PMID- 15777646 TI - Sequence comparison between the extracellular domain of M2 protein human and avian influenza A virus provides new information for bivalent influenza vaccine design. AB - To prevent the human and economic losses caused by human and avian influenza viruses, it is necessary to prepare safe bivalent influenza vaccines. Recent studies found that human influenza vaccines based on the extracellular domain of influenza M2 protein (M2e) induced broad-spectrum protective immunity in various antigen constructs. A prerequisite for using the M2e protein as a bivalent influenza vaccine component was to find out the sequence differences between human and non-human (avian or swine) influenza M2e proteins. Here, we completed such a comparison using 716 influenza M2e sequences available in Genbank. The results found one region on M2e protein consistent with host restriction specificities: PIRNEWGCRCN, PTRNGWECKCS and PIRNGWECRCN (aa10-20; the human, avian and swine specific M2e sequence, respectively). Interestingly, the comparison result was then validated by immunoblotting and enzyme-linked immunosorbent assay. The monoclonal antibody against the EVETPIRN sequence (aa6 13) of human M2e protein could weakly recognize avian M2e proteins bearing the EVETPTRN sequence (aa6-13) but failed to recognize avian M2e proteins bearing the EVETLTRN sequence (aa6-13). The data in this study provided useful information in the race to develop bivalent influenza vaccines against avian and human influenza A virus infection in human beings. PMID- 15777647 TI - Microarray and real-time RT-PCR analyses of differential human gene expression patterns induced by severe acute respiratory syndrome (SARS) coronavirus infection of Vero cells. AB - Vero E6 African green monkey kidney cells are highly susceptible to infection with the newly emerging severe acute respiratory syndrome coronavirus (SARS-CoV), and they are permissive for rapid viral replication, with resultant cytopathic effects. We employed cDNA microarray analysis to characterize the cellular transcriptional responses of homologous human genes at 12 h post-infection. Seventy mRNA transcripts belonging to various functional classes exhibited significant alterations in gene expression. There was considerable induction of heat shock proteins that are crucial to the immune response mechanism. Modified levels of several transcripts involved in pro-inflammatory and anti-inflammatory processes exemplified the balance between opposing forces during SARS pathogenesis. Other genes displaying altered transcription included those associated with host translation, cellular metabolism, cell cycle, signal transduction, transcriptional regulation, protein trafficking, protein modulators, and cytoskeletal proteins. Alterations in the levels of several novel transcripts encoding hypothetical proteins and expressed sequence tags were also identified. In addition, transcription of apoptosis-related genes DENN and hIAP1 was upregulated in contrast to FAIM. Elevated Mx1 expression signified a strong host response to mediate antiviral resistance. Also expressed in infected cells was the C-terminal alternative splice variant of the p53 tumor suppressor gene encoding a modified truncated protein that can influence the activity of wild type p53. We observed the interplay between various mechanisms to favor virus multiplication before full-blown apoptosis and the triggering of several pathways in host cells in an attempt to eliminate the pathogen. Microarray analysis identifies the critical host-pathogen interactions during SARS-CoV infection and provides new insights into the pathophysiology of SARS. PMID- 15777648 TI - Murine infection model for Mycobacterium fortuitum. AB - Mycobacterium fortuitum is an atypical, non-tubercular, pathogenic, rapidly growing mycobacteria. As very little is known about its virulence determinants, the absence of an animal infection model was always sorely felt. A reliable and reproducible murine infection model has been developed in which non-replicating persistence of 10(5) CFU/g tissue in kidney was observed when a standardised dosage inoculum of 5x10(7) CFU was injected intravenously. The tissue bacillary load was determined at regular intervals (10, 25, 45 and 60 days post inoculation) in different organs, viz., kidney, spleen, lung and liver. Histopathology of kidney revealed tissue damage and granuloma-like formations which appear to be part of the host's effort to combat the infection. As IFN gamma is known to trigger antimycobacterial effects of murine macrophages, IFN gamma was assayed to determine the correlation between host protective measures and bacillary load in kidney. Fifteen days after infection, the level of IFN gamma secreted by CD4+ and CD8+ T lymphocytes was high, concomitant with high tissue bacillary load, while the level sharply declined as the number of bacilli in kidney decreased 45 days post-inoculation. The invasion and proliferation of M. fortuitum ATCC 6841, when incubated with non-phagocytic (recombination activating genes (RAG) murine kidney) and phagocytic (murine peritoneal macrophages) cell lines, was assessed. M. fortuitum did not invade RAG murine kidney cell line, while the bacilli infected and proliferated freely inside murine macrophages. In conclusion, we present a reproducible murine infection model that sustains a persistent infection, the progression of which correlates meaningfully with host protective response. PMID- 15777649 TI - Impact of daily high-dose IFNalpha-2b plus ribavirin combination therapy on reduction of ALT levels in patients with chronic hepatitis C with genotype 1 and high HCV RNA levels. AB - The possibility of delaying progression to hepatocellular carcinoma in chronic hepatitis C patients with genotype 1 and high viral titers with baseline ALT levels of >/=50IU/L was examined by administration of IFN plus ribavirin combination therapy using ALT normalization as index and IFN monotherapy as control. The rate of sustained ALT normalization (ALT normal at 24 weeks after the end of treatment) was 28.1% with combination therapy and 10.5% with IFN monotherapy (P=0.001). Furthermore, the number of patients with sustained viral response (SVR) and with sustained ALT normalization in non-SVR patients was also significantly higher in the combination therapy versus monotherapy group. Mean ALT values during treatment and for 6 months after the end of treatment were significantly lower with combination therapy versus monotherapy even in virological nonresponders, as well as significantly lower during the post treatment observation period in patients who relapsed after the end of treatment. Since increase in the rate of sustained ALT normalization and SVR were successfully achieved, inhibition of progression to hepatocellular carcinoma should be studied with long-term IFN and ribavirin combination therapy. PMID- 15777650 TI - Extracellular nucleotides activate Runx2 in the osteoblast-like HOBIT cell line: a possible molecular link between mechanical stress and osteoblasts' response. AB - Dynamic mechanical loading increases bone density and strength and promotes osteoblast proliferation, differentiation and matrix production, by acting at the gene expression level. Molecular mechanisms through which mechanical forces are conversed into biochemical signalling in bone are still poorly understood. A growing body of evidence point to extracellular nucleotides (i.e., ATP and UTP) as soluble factors released in response to mechanical stimulation in different cell systems. Runx2, a fundamental transcription factor involved in controlling osteoblasts differentiation, has been recently identified as a target of mechanical signals in osteoblastic cells. We tested the hypothesis that these extracellular nucleotides could be able to activate Runx2 in the human osteoblastic HOBIT cell line. We found that ATP and UTP treatments, as well as hypotonic stress, promote a significant stimulation of Runx2 DNA-binding activity via a mechanism involving PKC and distinct mitogen-activated protein kinase cascades. In fact, by using the specific inhibitors SB203580 (specific for p38 MAPK) and PD98059 (specific for ERK-1/2 MAPK), we found that ERK-1/2, but not p38, play a major role in Runx2 activation. On the contrary, another important transcription factor, i.e., Egr-1, that we previously demonstrated being activated by extracellular released nucleotides in this osteoblastic cell line, demonstrated to be susceptible to both ERK-1/2 and p38 kinases. These data suggest a possible differential involvement of these two transcription factors in response to extracellularly released nucleotides. The biological relevance of our data is strengthened by the finding that a target gene of Runx2, i.e., Galectin 3, is up-regulated by ATP stimulation of HOBIT cells with a comparable kinetic of that found for Runx2. Since it is known that osteocytes are the primary mechanosensory cells of the bone, we hypothesize that they may signal mechanical loading to osteoblasts through release of extracellular nucleotides. Altogether, these data suggest a molecular mechanism explaining the purinoreceptors-mediated activation of specific gene expression in osteoblasts and could be of help in setting up new pharmacological strategies for the intervention in bone loss pathologies. PMID- 15777651 TI - Association and haplotype analyses of the COL1A2 and ER-alpha gene polymorphisms with bone size and height in Chinese. AB - Bone size (BS) is another risk factor of fracture independent of BMD in determining bone strength, and height is highly related with BS. To test the effect of the estrogen receptor-alpha (ER-alpha) and collagen type I alpha 2 (COL1A2) genes on the variation of BS and height, we genotyped the PvuII and XbaI polymorphisms in the intron 1 of the ER-alpha gene and the MspI and (GT)n markers in the intron 47 and intron 1 of the COL1A2 gene in 400 Chinese nuclear families with a total of 1256 individuals. The BS at the hip and spine was measured using a Hologic QDR 2000 dual-energy X-ray absorptiometry (DXA) scanner. Population stratification, total-family association, and within-family association were used to test the relationship of BS (at the spine and hip) and height with the four polymorphisms. We also performed these association analyses with the haplotypes of the MspI and (GT)n polymorphisms in the COL1A2 gene, and with the haplotypes of the PvuII and XbaI markers in the ER-alpha gene. Weak within-family association was found between the COL1A2-MspI (P = 0.05) and the femoral neck BS, between the ER-alpha-PX (P = 0.04) and the intertrochanter BS, and between the COL1A2-(GT)(17) (P = 0.02), COL1A2-m(GT)(17) (P = 0.009) and height. Subsequent permutation tests generally confirmed the suggestive within-family association. For the weak within-family association, the proportions of phenotypic variance accounted by the COL1A2-MspI, ER-alpha-PX, COL1A2-(GT)(17), COL1A2-m(GT)(17) markers were 1.50%, 1.51%, 2.15%, and 2.43% for the corresponding phenotypes. The association results indicate that the (GT)n and MspI markers of COL1A2 gene may have some influence on the variation of both BS and height, and the XbaI and PvuII markers of ER-alpha gene may have some effect on the variation of height in Chinese but not on the variation of BS. PMID- 15777652 TI - Limited nutritional energy supply differentially impairs growth and bone mineralization of the developing lumbar vertebrae in minipigs. AB - Bone development during puberty and adolescence may influence the later risk of osteoporosis. The aim of this study was to investigate whether the three dimensional development of the lumbar vertebrae is impaired in terms of bone mineral density and temporal control during the pubertal growth spurt when caloric intake is limited. Two groups of prepubertal female Gottingen minipigs (each n = 6) were fed either ad libitum (free fed) or with reduced food intake (controlled fed) for 12 months using a standard diet. Apparent volumetric bone mineral densities (vBMD) and size of the 4th vertebra were measured by quantitative CT in monthly intervals. The weight of free-fed animals was 29.5 +/- 0.9 kg compared to 25.9 +/- 3.5 kg in the controlled-fed group at the end of the observation period (mean +/- SD; p < 0.05). The maximal cross-sectional area of the 4th vertebra was 148.2 +/- 3.4 mm(2) in free-fed animals compared to 143.1 +/ 6.3 mm(2) in controlled-fed animals (P < 0.05) as both, the cortical and the medullary area were greater in the free-fed group. In contrast, cancellous and cortical bone mineral densities were not different between the two groups; however, maximal densities were reached earlier in the free-fed group. As regards the height of the vertebra, no difference was observed between the groups after 12 months (25.46 +/- 0.54 mm in controlled fed vs. 25.56 +/- 0.45 mm in free fed), although the peak height was reached earlier in the free-fed group. In both groups, the maximal growth rate in the cross-sectional direction was reached earlier compared to the longitudinal direction (both P < 0.05). Growth in the longitudinal direction that occurs in the primary and secondary epiphysis does not appear to depend on caloric intake contrary to the periosteal and endocortical bone generation in the cross-sectional direction. When limited energy is available, the organism seems to preserve the development of normal body height, while accepting reduced strength of the vertebra. PMID- 15777653 TI - Regional differences in cortical bone mineral density in the weight-bearing long bone shaft--a pQCT study. AB - This study used a multislice peripheral quantitative computed tomography (pQCT) to measure volumetric BMD (vBMD) and cortical thickness for investigating regional adaptation in lower tibial shaft in 72 healthy postmenopausal women aged 47-60. Tomographic slices were analysed on four distinct cortical regions: the anterior, posterior, medial and lateral cortical wall. One-way analysis of variance (ANOVA) test was used to compare the vBMD in the four regions. The results showed that the posterior cortex had the highest vBMD (1923 +/- 135.3 mg/cm(3)), significantly (P < 0.001) higher than the anterior cortex (1805 +/- 110.6 mg/cm(3)), medial cortex (1863 +/- 103.6 mg/cm(3)) and lateral cortex (1815 +/- 111.6 mg/cm(3)); whereas there was no significant difference (P > 0.05) between the medial and lateral cortices located near the neutral plane of bending. The anterior cortex had the greatest thickness (2.56 +/- 0.47 mm), significantly (P < 0.001) greater than that of the posterior cortex (2.11 +/- 0.27 mm), medial cortex (2.20 +/- 0.39 mm) and lateral cortex (2.03 +/- 0.29 mm). The vBMD of the posterior cortex was a significant 6.5% higher than that of the anterior cortex (P < 0.001); whereas the anterior cortical thickness was a significant 21.3% greater than that of the posterior cortex (P < 0.001). There was no linear relationship found between cortical vBMD and cortical thickness measured at the four cortical regions (r = 0.086, P > 0.05). In conclusion, the regional differences, with higher vBMD found in posterior cortex, might be a result of mechanical adaptation, which caused the posterior cortex to sustain higher compressive loading than the anterior tensile cortex during the landing phase in the gait cycles of individuals. Nevertheless, regional geometric adaptation in anterior cortical thickness might be adapted to accommodate for the reduced vBMD and to reduce the bending stress in this region. PMID- 15777654 TI - The hyalectan degrading ADAMTS-1 enzyme is expressed by osteoblasts and up regulated at regions of new bone formation. AB - During bone formation, there are numerous pivotal changes in the interrelationships between osteoblasts and molecules of the extracellular matrix (ECM). Consequently, the mechanisms that underlie the temporal and spatial distribution of ECM molecules in bone are of considerable interest in understanding its formation. A subfamily of a disintegrin and metalloproteinase (ADAMs) has been identified, which contain thrombospondin-like motifs (ADAMTS), and can break down several ECM molecules. Using reversed transcribed PCR, we identified ADAMTS-1, -4 and -5 mRNA expression in cultures of rat osteoblasts treated with ascorbic acid, beta-glycerophosphate and dexamethasone, molecules known to drive osteoblast differentiation. Of these, ADAMTS-1 followed most closely the osteogenic marker osteocalcin during in vitro mineralisation. Consequently, we studied, in detail, protein expression of ADAMTS-1 during in vitro osteogenesis together with ADAMTS-1 immunohistochemistry staining of sections from 2- and 10-day-old rat femur. Western analysis of osteoblast proteins showed ADAMTS-1 products that correspond well with both full-length and furin-processed species. In the ECM laid down by osteoblasts, only the mature secreted protein (approximately 90 kDa) and its accumulation during the later stages of osteogenesis in vitro were noticed. Furthermore, immunostaining with an antibody recognising ADAMTS-1 demonstrated strong expression around mineralised nodules and intense focal staining of putative new areas of nodule formation in vitro. Finally, immunohistochemistry of 2- and 10-day-old rat femur localised ADAMTS-1 protein to regions associated with osteogenesis. These data show that ADAMTS-1 protein accumulates in osteoblast ECM during differentiation. Furthermore, the focalised expression of ADAMTS-1 in regions of osteogenesis, both in vitro and in vivo, implicates this multifunctional protein to be involved in mineralised nodule and bone formation. PMID- 15777655 TI - Low dose fibroblast growth factor-2 (FGF-2) enhances bone morphogenetic protein-2 (BMP-2)-induced ectopic bone formation in mice. AB - To examine how fibroblast growth factor-2 (FGF-2) affects the BMP signaling pathway during bone morphogenetic protein-2 (BMP-2)-induced ectopic bone formation, we implanted type I collagen disks containing constant amounts of BMP 2 (5 micrograms) and varying amounts of FGF-2 onto the back muscles of adult male mice. We then performed histological analyses and histomorphometry, and measured bone mineral density and radiopaque area on the discs 1, 2, and 3 weeks after implantation. We also determined the expression profiles of several genes involved in bone formation and the BMP signaling pathway in the muscle that had been adjacent to the implanted disc and in muscle-derived primary culture cells that had similarly been treated with a constant concentration of BMP-2 and a varying concentration of FGF-2. In the presence of a constant amount of BMP-2, we confirmed that low doses of FGF-2 increased ectopic bone formation in vivo and high doses inhibited bone formation. Northern and/or Western blots of recovered muscle from the in vivo experiment and treated muscle-derived primary culture cells from the in vitro experiment revealed that low doses of FGF-2, but not high doses, increased the expression BMP receptor (BMPR)-1B, phosphorylated Smad1, Noggin, and Osteocalcin. Our results indicate that low-dose FGF-2 may facilitate BMP-2-induced ectopic bone formation by altering the expression of BMPRs on the surface of bone forming progenitor cells. They also indicate that the inhibitory effect of high-dose FGF-2 is not mediated via increased expression of the BMP inhibitor Noggin. PMID- 15777656 TI - Prediction of fracture callus mechanical properties using micro-CT images and voxel-based finite element analysis. AB - Assessment of fracture healing is a common problem in orthopaedic practice and research. To determine the effectiveness of certain treatments, drugs, mechanical loads, or rehabilitation regimes, the strength of the fracture callus must be determined. Both clinically and experimentally, there is a need to noninvasively and quantitatively evaluate fracture callus quality during healing. The objective of this study was to develop a method to assess fracture stiffness using micro computed tomography (micro-CT) and finite element analysis. The method was developed and validated with plastic phantoms of various cross sections and known material properties, tested experimentally in four-point bending and torsion. The method was then applied to fractured rat femurs after 3 and 4 weeks of healing tested experimentally in torsion (50 femurs total). Micro-CT scans were made of the fracture calluses to determine three-dimensional geometry and material properties for the finite element models. Experimentally measured torsional rigidities were compared to finite element solutions. Finite element model predictions of callus rigidity correlated significantly better with experimental torsional rigidity than other common measures of healing progress such as callus area, bone mineral density, or area moment of inertia. Using FEA to predict mechanical properties of the callus could prove to be a useful tool in fracture healing studies. PMID- 15777657 TI - The C terminus of MINT forms homodimers and abrogates MINT-mediated transcriptional repression. AB - Notch signaling plays a pivotal role in numerous cell fate determination events during development, and therefore its regulation has been studied intensively. MSX2-interacting nuclear target protein (MINT) modifies the Notch signaling by interacting with and inhibiting the downstream transcription factor RBP-J/CBF-1 of Notch. In this study, by a yeast two hybrid screening, we found that the C terminal fragment of MINT interacted with each other. We confirmed the interaction between two MINT C terminal fragments both in vitro and in vivo. We further demonstrated that the overexpression of the C terminal fragment of MINT cancelled its inhibitory effect on the transactivation of an RBP-J-dependent promoter by Notch. These results suggest that MINT may form a dimer or multimer in cells through its C terminus, and that the C terminal fragment of MINT may work as its dominant-negative version. PMID- 15777658 TI - The complete genomic sequence of the carp fast skeletal myosin heavy chain gene. AB - We have determined the complete DNA nucleotide sequence of the carp Cyprinus carpio fast skeletal myosin heavy chain (MYH) gene. Introns and exons were predicted by comparison with the corresponding carp MYH cDNAs previously reported. The gene encoded the entire mRNA transcript and contained 5958 nucleotides (nt) including 77 nt 5'-untranslated region, 5796 nt coding region for 1931 amino acids, and 85 nt 3'-untranslated region. The coding region was split by 38 introns and the complete gene contained 11,385 nt. This integration of the carp fast skeletal MYH gene was comparable to those of the rat and chicken embryonic MYH genes, which have 41 and 40 exons, respectively. However, the entire gene size of carp MYH was about half those of rat and chicken due to much smaller size of carp introns. We have also demonstrated that this carp MYH gene belonged to so-called intermediate type in a multigene family of carp fast skeletal muscle MYH in comparison of its nucleotide and deduced amino acid sequences to those of carp MYH cDNAs reported previously. PMID- 15777659 TI - Sp1: regulation of gene expression by phosphorylation. AB - As the prototype of a family of transcription factors, Sp1 has been extensively studied and widely reported for its role in gene regulation. The first evidence of Sp1 phosphorylation was reported more than a decade ago. Since then, an increasing number of Sp1 phosphorylation events have been characterized. Recent data demonstrate an important role for the phosphorylation state of Sp1 in the regulation of multiple genes. In this article, we review published literature in four specific areas relating to the phosphorylation of Sp1: (1) signal transduction pathways for Sp1 phosphorylation, (2) mechanisms of Sp1 dephosphorylation, (3) the functional implications of Sp1 phosphorylation, and (4) Sp1 phosphorylation in the lung. PMID- 15777660 TI - Sexuality and parasitism share common regulatory pathways in the fungus Parasitella parasitica. AB - Parasitella parasitica, a facultative mycoparasite of zygomycetous fungi, forms cytoplasmic fusions with its hosts during infection. Thus, the organism is an efficient donor of genetic material in parasexual host-parasite interactions. Recognition between parasite and host is mediated by trisporoids, which are also responsible for sexual communication. The TDH gene for one of the key enzymes of trisporic acid biosynthesis, 4-dihydromethyl-trisporate dehydrogenase, was cloned and its transcription analysed. TDH was cloned on a 6175-bp insert and was found to map in a complex cluster of genes that suggest post-transcriptional antisense regulation. Histochemical TDH analysis in developing parasitic or sexual structures shows high enzymatic activity in Parasitella. TDH is linked to a gene for a putative acyl-CoA thioesterase (ACT). Two ORFs were identified in the 5' region of the TDH gene, a third one, coding for 176 amino acids overlaps the ACT gene in antisense direction completely. Expression levels of ACT and ORF1 depend on parasitic and sexual interactions. PMID- 15777661 TI - Involvement of the "linker" region between the exonuclease and polymerization domains of phi29 DNA polymerase in DNA and TP binding. AB - For several DNA-dependent DNA polymerases it has been shown that their synthetic and degradative activities are organized in two separated modules. The functional coordination required between them to accomplish successfully the replication process is provided by important contacts with the substrate contributed by residues coming from both modules. These domains are connected by a central "linker" region adjacent to the "YxGG/A" motif, the putative limit of the polymerization domain. We describe here the mutational analysis of phi29 DNA polymerase in several residues of this region, connecting the N- and C-terminal domains and conserved in DNA polymerases able to start replication by protein priming. The mutant polymerases with the less conservative changes showed reduced DNA binding activity. Additionally, their TP binding capacity was reduced, affecting the TP-deoxynucleotidylation in the absence of template. Interestingly, the role of the residues studied here in DNA binding seems to be especially important to start replication, when the polymerase enters from the closed binary into the ternary complex. These results allow us to propose that this interdomain region of phi29 DNA polymerase is playing an important role for substrate binding including both DNA and TP. PMID- 15777662 TI - Length variation of CAG/CAA triplet repeats in 50 genes among 16 inbred mouse strains. AB - CAG repeats coding for poly-glutamines have been studied by many groups as repeat length variations contributes to differences in protein function and disease outcome. In this study, we systematically searched public databases for genes carrying CAG repeats. For the genes obtained, we experimentally analyzed variations of length and the purity of the repeats in 62 loci among 16 inbred mouse strains, including wild-derived and laboratory strains. We found that length was conserved in 50% of the loci, especially among wild-derived strains. Of 496 polymorphic repeat alleles, 78% were uninterrupted and 22% were interrupted with non-CAG codons. Interruptions tended to occur in longer repeats and all repeats of greater length than 23 were interrupted. Although interruptions can act as suppressors for the expansion of CAG repeats, we found that the occurrence of the interruptions depended on the length of the CAG repeats. Furthermore, most poly-glutamines examined in this study existed in human orthologous genes, reflecting the functional significance of poly glutamines in proteins. PMID- 15777663 TI - Molecular cloning and characterization of a cellulase gene from a symbiotic protist of the lower termite, Coptotermes formosanus. AB - The endo-beta-1,4-glucanase gene was cloned from a cDNA library constructed from the mixed population of symbiotic protists in the hindgut of the lower termite, Coptotermes formosanus, using the lambda ZAP II vector. The recombinant phage library was screened for cellulolytic activity by the Congo red staining procedure. The nucleotide sequence comprised 941 nucleotides including a polyA tail sequence and showed high sequence similarity with endoglucanase genes belonging to glycosyl hydrolase family 5. Determination of the 5' end of the cellulase gene using the 5'RACE method showed that the full-length cDNA comprised a 921-bp ORF, encoding a putative 33,620 Da protein. The organismal source of this cellulase gene was identified using PCR with gene-specific primers and whole cell in situ hybridization as the smallest symbiotic hypermastigote protist, Spirotrichonympha leidyi. The optimal pH and temperature of the cellulase heterologously expressed in Escherichia coli were 5.8-6.0 and 70 degrees C, respectively. The Km and Vmax values on carboxymethyl cellulose (CMC) substrate were 1.90 mg/ml and 148.2 units/mg protein, respectively. PMID- 15777664 TI - WITHDRAWN: Molecular evolution of a microRNA cluster in the PWS/AS region among mammals. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 15777665 TI - The mitochondrial genome of Indonesian coelacanth Latimeria menadoensis (Sarcopterygii: Coelacanthiformes) and divergence time estimation between the two coelacanths. AB - We determined the whole mitochondrial genome sequence for Indonesian coelacanth Latimeria menadoensis. The genome content and organization were identical to that of typical vertebrates including Comoran coelacanth, Latimeria chalumnae. The overall nucleotide differences between the two species (excluding the control region) was 4.28%. The divergence time between the two species was estimated using whole mitochondrial genome data from the two coelacanths and 26 actinopterygians that represent major actinopterygian lineages plus an outgroup. Partitioned Bayesian analyses were conducted with the two data sets that comprised concatenated amino acid sequences from 12 protein-coding genes (excluding ND6 gene) and concatenated nucleotide sequences from 12 protein-coding genes (without 3rd codon positions), 22 transfer RNA genes, and two ribosomal RNA genes. The molecular clock analysis was also conducted with the concatenated amino acid sequences from the 12 protein-coding genes after removing faster or more slowly evolving sequences. Using the sarcopterygian-actinopterygian split as a calibration point (450 Mya), divergence time estimation between L. menadoensis and L. chalumnae fell in the range of 40-30 Mya, which is much older than those of the previous studies (<6.3 Mya). Assuming that the most recent ancestor of Latimeria was distributed continuously along the deep coasts of Africa through Eurasia, our estimate is in agreement with the hypothesis that the collision of India with Eurasia (50 Mya) and the subsequent siltation caused by the formation of major rivers resulted in a coelacanth habitat disjunction that allowed populations on either side of India to diverge. PMID- 15777666 TI - Patterns of dentate granule cell responses to perforant path stimulation in epileptic mice with granule cell dispersion. AB - In adult mice, intrahippocampal administration of kainic acid induces a structural modification of the granule cell layer reminiscent of granule cell dispersion (GCD) seen in humans with temporal lobe epilepsy. We tested that GCD might be involved in the patterns of granule cell responses to perforant path stimulation by recording field potentials in vivo after kainic acid-induced status epilepticus until the phase of chronic seizure activity in presence of GCD or after its alteration by K252a co-treatment, an inhibitor of tyrosine kinase activities. Stimulation triggered bursts of multiple population spikes, the number of which progressively increased with time whereas their amplitude decreased in parallel with the progressive decrease in granule cell density. The population spike threshold was reached for a lower excitatory synaptic drive than in controls, as assessed by the initial slope of the field excitatory post synaptic potential. This indicates that, for identical synaptic responses, granule cells were closer to the firing threshold. Fast inhibition, assessed by paired pulse stimulation, was compromised immediately after the initial status epilepticus, consistent with the rapid loss of most hilar cells. Neither the epileptic course nor the epileptiform responses of the granule cells were modified and manipulation by alteration following GCD manipulation while granule cell neuropeptide-Y immunostaining was substantially decreased. In this mouse model of TLE, granule cells display a progressive increase in epileptiform responses to afferent input until the occurrence of spontaneous seizures. The population spike amplitude decreases in parallel with GCD while the granule cell excitability is enhanced. Consequently, data from field potentials in epilepsy experiments should be interpreted with care, taking into account the possible variations in the neuronal density in the recorded area. PMID- 15777667 TI - Antiviral reactivities of gammadelta T cells. AB - The complex antiviral immune mechanisms involve both adaptive and innate reactions mediated by gammadelta T lymphocytes, whose unique immunosurveillance contributions are analyzed here in different clinical and experimental settings. It is beyond any doubt that the fast, potent, cytotoxic as well as non-cytolytic antiviral activities of gammadelta T cells are critical in protecting the host against diverse viral pathogens. PMID- 15777668 TI - Ventilator-associated pulmonary infection: the germ theory of disease remains viable. AB - Pulmonary infection complicating mechanical ventilation is a major problem in critical care. The key issues surrounding care of patients suspected of having this disease are 1) appropriate diagnostic criteria; 2) when antibiotic therapy should be started; and 3) what constitutes adequate antibiotic therapy. Current data support use of quantitative cultures obtained by either bronchoscopic or blind catheter lavage or mini-brushing. Antibiotic therapy should be guided by duration of hospitalization prior to presumed infection and local predominating nosocomial organisms and their microbial resistance patterns. The key issue with timing of therapy now centers around early termination of therapy if quantitative cultures are negative. PMID- 15777669 TI - Oncogenic osteomalacia: diagnostic importance of fibroblast growth factor 23 and F-18 fluorodeoxyglucose PET/CT scan for the diagnosis and follow-up in one case. AB - A case of oncogenic osteomalacia is reported in a 71-year-old man who presented with bone pain, muscle weakness, and severe hypophosphatemia. The tumor which was localized in the left lower mandible was not detected by tomodensitometry, resonance magnetic imaging, and (111)IN-octreotide scintigraphy, but was easily localized by F-18 fluorodeoxyglucose PET/CT SCAN (F-18 FDG PET/CT SCAN). To our knowledge, the value of this technique for detecting tumors in oncogenic osteomalacia has never been reported. Secondly, this case provided an opportunity for confirming the usefulness of serum fibroblast growth factor 23 (FGF23) measurement for the diagnosis and follow-up. We conclude that FGF23 measurements combined with F-18 FDG PET/CT SCAN were decisive tools in a case of oncogenic osteomalacia and are likely to be of considerable importance for facilitating early diagnosis and follow-up in the future. PMID- 15777670 TI - An intermediate form of juvenile Paget's disease caused by a truncating TNFRSF11B mutation. AB - Juvenile Paget's disease (JPD) is a rare condition with an autosomal recessive mode of inheritance. Typically presenting in infancy or early childhood, the disorder is characterized by a generalized widening of the long bones and thickening of the skull combined with sustained elevation of serum alkaline phosphatase levels. The extremely rapid bone turnover results in osteopenia, fractures, and progressive skeletal deformity. In 2002, mutations in TNFRSF11B, the gene encoding osteoprotegerin, were described as underlying JPD. We evaluated a patient with JPD at the clinical, biochemical, radiological, and molecular level. Mutation analysis of TNFRSF11B revealed a homozygous insertion/deletion in exon 5, predicted to result in truncation of the protein at amino acid 325. The residual activity of the mutated protein product was investigated by Western blotting and ELISA upon transient overexpression. Absence of the C-terminal domain abolished homodimerization and was shown to lead to a decreased capacity of the mutant protein to bind its ligand RANKL. We conclude that truncation of the C-terminal part of osteoprotegerin negatively affects functional activity. As a consequence, osteoclast formation and function are up-regulated, causing the increased bone turnover seen in this patient. PMID- 15777671 TI - Effects of hyaluronan on three-dimensional microarchitecture of subchondral bone tissues in guinea pig primary osteoarthrosis. AB - Hyaluronan (HA) has received increasing interest as a potential agent in therapeutic intervention in osteoarthrosis (OA). HA has been shown to reduce arthritic lesions in experimental animal models of articular cartilage injury. This study was to investigate the effects of high molecular weight HA intra articular injection on subchondral bone tissues. Fifty-six male guinea pigs were randomly divided into 5 groups. During the initial 2.5-month period, three groups received intra-articular injection of HA 0.4 mg/kg/week for 5 weeks in both knee joints. Two control groups received vehicle. After 2.5 months, one HA group and one control group were sacrificed. The remaining 3 groups (5.5-month groups) were left for an additional 3 months before sacrifice during which time one HA group received additional 5 weeks injections, one HA group received no more injections, and the control group received vehicle. The left tibiae were harvested and micro CT scanned to quantify three-dimensional microarchitecture of subchondral bone plate, cancellous bone and cortical bone, followed by mechanical testing and collagen and mineral determinations. All HA-treated groups had almost normal cartilage, whereas the control groups had typical OA-related cartilage degradation. In the 2.5-month group, HA resulted in significantly decreased subchondral plate volume fraction and thickness and HA-treated cancellous bone had significantly lower bone volume fraction, and typical rod-like structure. After 5.5 months, these changes were more pronounced, with an additional marked decrease in connectivity and bone surface density. HA-treated cortical bone had significantly greater volume fraction at both observation times. HA groups had greater bone mineral concentration and reduced collagen to mineral ratio with similar mechanical properties of cancellous bone but less stiff cortical bone. The effects of HA on cartilage and subchondral bone were maintained when treatment was discontinued. In summary, HA effectively protects against cartilage degeneration, decreases subchondral bone density and thickness, changes trabecular structure toward rod-like, so that subchondral bone becomes more compliant and thereby reduces cartilage stress during impact loading. HA preserved cancellous bone mechanical properties by increasing bone mineralization. Early HA administration is effective for intervention of OA initiation and progression, and short-term early HA treatment is sufficient to maintain treatment effects. PMID- 15777672 TI - Inducible nitric oxide synthase mediates bone development and P. gingivalis induced alveolar bone loss. AB - The role of inducible nitric oxide synthase (iNOS) in bone development and bacterially induced periodontal bone loss was examined using mice with targeted mutation of the iNOS gene. Femurs of iNOS KO mice showed 30% and 9% higher bone mineral density compared to wild type (WT) at 4 and 9 weeks of age, respectively. Micro-computed tomography revealed that cortical thickness and cortical bone density is increased in the absence of iNOS, while trabecular bone thickness and bone density remains unchanged. Histochemical analysis using TRAP staining showed that osteoclast numbers are lower by 25% in iNOS KO femurs compared to WT femurs. When bone marrow cells were stimulated with M-CSF and RANKL in vitro, iNOS KO cultures developed 51% fewer TRAP-positive multinuclear cells compared to WT cultures. When similar cultures were grown on dentine discs, resorption pit area was decreased by 54% in iNOS KO cultures. Gene expression studies showed that iNOS expression is induced by M-CSF and RANKL in WT bone marrow cultures, while no iNOS transcript was detected in iNOS KO. No compensatory change was detected in the expression of neuronal or endothelial NOS isoforms. There was no difference in RANK and osteoprotegerin expression between iNOS KO and WT bone marrow cultures after M-CSF and RANKL-treatment, while Traf6 expression was significantly lower in the absence of iNOS. In the alveolar bone of the maxilla, the distance between the cementoenamel junction and the alveolar bone crest was larger in iNOS KO compared to WT mice from 6 to 14 weeks of age, indicating a developmental effect of iNOS in oral tissues. Oral administration of the periodontal pathogen Porphyromonas gingivalis caused alveolar bone loss in the maxilla of WT mice, but failed to do so in iNOS KO mice. Expression of the osteoclast marker cathepsin K was 25% lower in iNOS KO alveolar bone. These data indicate that iNOS promotes bone resorption during bone development as well as after bacterial infection, and that iNOS is an important signal for normal osteoclast differentiation. PMID- 15777673 TI - Low BMD is less predictive than reported falls for future limb fractures in women across Europe: results from the European Prospective Osteoporosis Study. AB - We have previously shown that center- and sex-specific fall rates explained one third of between-center variation in upper limb fractures across Europe. In this current analysis, our aim was to determine how much of the between-center variation in fractures could be attributed to repeated falling, bone mineral density (BMD), and other risk factors in individuals, and to compare the relative contributions of center-specific BMD vs. center-specific fall rates. A clinical history of fracture was assessed prospectively in 2451 men and 2919 women aged 50 80 from 20 centers participating in the European Prospective Osteoporosis Study (EPOS) using standardized questionnaires (mean follow-up = 3 years). Bone mineral density (BMD, femoral neck, trochanter, and/or spine) was measured in 2103 men and 2565 women at these centers. Cox regression was used to model the risk of incident fracture as a function of the person-specific covariates: age, BMD, personal fracture history (PFH), family hip fracture history (FAMHIP), time spent walking/cycling, number of 'all falls' and falls not causing fracture ('fracture free') during follow-up, alcohol consumption, and body mass index. Center effects were modeled by inclusion of multiplicative gamma-distributed random effects, termed center-shared frailty (CSF), with mean 1 and finite variance theta (theta) acting on the hazard rate. The relative contributions of center-specific fall risk and center-specific BMD on the incidence of limb fractures were evaluated as components of CSF. In women, the risk of any incident nonspine fracture (n = 190) increased with age, PFH, FAMHIP, > or =1 h/day walking/cycling, and number of 'all falls' during follow-up (all P < 0.074). 'Fracture-free' falls (P = 0.726) and femoral neck BMD did not have a significant effect at the individual level, but there was a significant center-shared frailty effect (theta = 0.271, P = 0.001) that was reduced by 4% after adjusting for mean center BMD and reduced by 19% when adjusted for mean center fall rate. Femoral trochanter BMD was a significant determinant of lower limb fractures (n = 53, P = 0.014) and the center-shared frailty effect was significant for upper limb fractures (theta = 0.271, P = 0.011). This upper limb fracture center effect was unchanged after adjusting for mean center BMD but was reduced by 36% after adjusting for center mean fall rates. In men, risk of any nonspine fracture (n = 75) increased with PFH, fall during follow-up (P < 0.026), and with a decrease in trochanteric BMD [RR 1.38 (1.08, 1.79) per 1 SD decrease]. There was no center effect evident (theta = 0.081, P = 0.096). We conclude that BMD alone cannot be validly used to discriminate between the risk of upper limb fractures across populations without taking account of population-specific variations in fall risk and other factors. These variations might reflect shared environmental or possibly genetic factors that contribute quite substantially to the risk of upper limb fractures in women. PMID- 15777674 TI - A PPAR response element regulates transcription of the gene for human adipose differentiation-related protein. AB - Lipid droplets are cytoplasmic organelles which serve as storage sites for neutral lipids. Adipose differentiation-related protein (ADRP) is intrinsically associated with the surface of lipid droplets and is believed to play a major role in the maintenance of lipid stores in non-adipocytes. ADRP abundance is intimately linked to the amount of lipid found within cells and agents which increase the levels of intracellular lipid, such as certain agonists of the peroxisome proliferator-activated receptors (PPARs), also are capable of modulating ADRP gene transcription. However, little is known about the molecular mechanisms and promoter control elements, which regulate the transcription of the human gene. Using a reporter system to investigate ADRP transcription, we have identified a PPAR response element (PPRE) with the sequence 5'-AGGTGA A AGGGCG-3' within its promoter region. Mutational analysis revealed that the ADRP PPRE specifically mediated the upregulation of transcription in response to activation by agonists of PPAR subtypes alpha and delta in both rat and human hepatocyte derived cell lines. These findings offer insight into the mechanisms which serve to regulate ADRP transcription and intracellular lipid storage. PMID- 15777675 TI - Structural characterization and transcriptional regulation of the gene encoding diapause hormone and pheromone biosynthesis activating neuropeptide in the cotton bollworm, Helicoverpa armigera. AB - We have cloned the gene encoding the diapause hormone and the pheromone biosynthesis activating neuropeptide in Helicoverpa armigera (Har-DH-PBAN). The Har-DH-PBAN gene contains six exons and five introns that fall in the same positions as in the Bombyx mori DH-PBAN gene (Bom-DH-PBAN). The transcription initiation site lays 29 bp upstream of the translation initiation site. Southern blot analysis suggests that a single copy of this gene is present per haploid genome. A structural comparison of DH-PBAN promoters between H. armigera and B. mori show similarities in the TATA box and in a potential binding site for a POU family transcription factor, POU-M2. However, testing of these DNA regions for factor binding in vitro and transcription assays in cell culture highlight significant differences in their regulation particularly in reference to the POU M2 sites. Our results uncover common and different regulatory mechanisms at work in the control of DH-PBAN gene expression in H. armigera and B. mori. PMID- 15777676 TI - Evolutionary turnover of two pBuM satellite DNA subfamilies in the Drosophila buzzatii species cluster (repleta group): from alpha to alpha/beta arrays. AB - The pBuM satellite DNA family was studied in seven Drosophila species from the buzzatii cluster (within the large Drosophila repleta group). The pBuM repeats are slightly AT-rich and show high levels of intraspecific sequence homogeneity. The pBuM family can be divided into two subfamilies. The pBuM-1 subfamily consists of tandemly arranged repetition units of approximately 190 bp, termed alpha. Alpha repeats were found in a high copy number in the genome of D. buzzatii, D. serido and D. antonietae. The pBuM-2 subfamily consists of tandemly arranged repetition units of 370 bp. Its origin is explained by an insertion of an approximately 180 bp foreign sequence (termed beta) in an alpha basic repeat unit, with subsequent homogenization/amplification events increasing its frequency. Alpha/beta repeats were found in a high copy number in the genome of D. serido, D. antonietae, D. seriema and D. gouveai. pBuM sequences were not detected in D. koepferae and D. borborema by hybridization experiments. The nucleotide analysis of 74 pBuM repeats revealed that apart from the beta insertion event, the evolution of the pBuM family has proceeded in a gradual fashion, mainly through accumulation and horizontal spread of nucleotide substitutions. Moreover, the data also indicate a faster evolutionary rate for the pBuM-2 subfamily than the pBuM-1 subfamily. Members of both subfamilies display a greater intraspecific than interspecific homogeneity, indicating a concerted mode of pBuM evolution. A scenario to explain the evolution of both satDNA subfamilies in the seven Drosophila species from the buzzatii cluster is proposed. PMID- 15777677 TI - Isolation and characterization of the Bactrocera oleae genes orthologous to the sex determining Sex-lethal and doublesex genes of Drosophila melanogaster. AB - Here we report the isolation and characterization of the olive fruit fly Bactrocera oleae genes orthologous to the Drosophila melanogaster sex-determining genes Sex-lethal (Sxl) and doublesex (dsx). Fragments of the Sxl and dsx orthologous were isolated with RT-PCR. Genomic and cDNA clones were then obtained by screening a genomic library and separate male and female cDNA adult libraries using the RT-PCR products as probes in both cases. B. oleae Sxl gene (BoSxl) expresses the same pattern of transcripts which encode for a single common polypeptide in both male and female flies. The gene shares a high degree of similarity in sequence and expression to its Ceratitis capitata orthologous and does not appear to play a key regulatory role in the sex-determining cascade. B. oleae dsx gene (Bodsx) expands in a chromosomal region of more than 50 kb, with 6 exons-5 introns, producing different sex-specific mRNAs, according to the Drosophila model. The cDNA sequences are almost identical to the gene orthologous of Bactrocera tryoni. Four repeat elements identical to the D. melanogaster TRA/TRA-2 binding sites have been found in the untranslated region of the female specific exon 4, predicting a common regulatory splicing mechanism in all studied species of Diptera. PMID- 15777678 TI - Effect of a selective agonist for prostaglandin E receptor subtype EP4 (ONO-4819) on the cortical bone response to mechanical loading. AB - The influence of a selective agonist for prostaglandin E receptor subtype EP4 (ONO-4819) on the bone response to mechanical loading was evaluated. Six-month old female Wistar rats were used and assigned to three groups (n = 12/group): Vehicle administration (EP4-V), low-dose ONO-4819 administration (EP4-L, 3 microg/kg BW), and high-dose ONO-4819 administration (EP4-H, 30 microg/kg BW). ONO-4819 was subcutaneously injected in the back twice a day for 3 weeks. Loads on the right tibia at 39.4 N for 36 cycles at 2 Hz were applied in vivo by 4 point bending every other day for 3 weeks. Whole-body bone mineral content showed a significant difference between EP4-V and EP4-H (P < 0.05). Bone mineral density (BMD) of the total and regional tibia (the region with maximal bending at the central diaphysis) was higher in EP4-H than EP4-V, showing a significant effect of loading (P < 0.001) and ONO-4819 (P < 0.05). BMD of the total femur was higher in EP4-H than EP4-V (P < 0.01) and that of the distal femur was higher in EP4-H than EP4-V (P < 0.001). Histomorphometry of the cortical bone showed that loading increased formation surface (FS/BS), mineral appositional rate (MAR), and bone formation rate (BFR/BS) significantly at the lateral periosteal surface (P < 0.001); however, the effect of ONO-4819 was not significant. At the medial periosteal surface, loading increased the three parameters (P < 0.001) and ONO 4819 increased FS/BS (P < 0.001) and MAR (P < 0.05) significantly. At the endocortical surface, the effects of both loading and ONO-4819 were significant on all three parameters (for loading; FS/BS P < 0.01, MAR P < 0.05, BFR/BS P < 0.03, for ONO-4819 all P < 0.001). It was concluded that ONO-4819 increased cortical bone formation in rats and there was an additive effect on the bone response to external loading by 4-point bending. PMID- 15777679 TI - Mechanosensitivity of the rat skeleton decreases after a long period of loading, but is improved with time off. AB - After the initial adaptation to large mechanical loads, it appears as though the skeleton's responsiveness to exercise begins to wane. To counteract the waning effects of long-term mechanical loading, "time off" may be needed to improve the responsiveness of bone cells to future mechanical signals and reinitiate bone formation. The aim of this study was to determine whether bone becomes less sensitive to long-term mechanical loading and whether time off is needed to improve mechanosensitivity. Fifty-seven female Sprague-Dawley rats (7-8 months of age) were randomized to one of following groups: Group 1 loading was applied for 5 weeks followed by 10 weeks of time off (1 x 5); Group 2 loading was applied for 5 weeks, followed by time off for 5 weeks and loading again for 5 weeks (2 x 5); Group 3 loading was applied continuously for 15 weeks (3 x 5); Group 4 age matched control group; and Group 5 baseline control group. An axial load was applied to the right ulna for 360 cycles/day, at 2 Hz, 3 days/week at 15 N. At the end of the intervention, all three loaded groups showed similar increases in bone mass, cortical area, and I(MIN) in response to mechanical loading(.) Bone formation rate of the loaded ulna was increased in the first 5 weeks of loading for all three loaded groups; however, during the last 5 weeks, it was only significantly increased in the group that had time off (2 x 5) (P < 0.05). The group that had time off (2 x 5) also showed greater improvements in work to failure compared to the group loaded for 5 weeks (1 x 5) and the entire 15 weeks (3 x 5). A second experiment showed that the waning effect of long-term loading on the skeleton is not a result of aging. In conclusion, mechanical loading of the rat ulna results in large improvements in bone formation during the first 5 weeks of loading, but continual loading decreases the osteogenic response. Having time off increases bone formation and improves the resistance to fracture. PMID- 15777680 TI - Are effects of MTHFR (C677T) genotype on BMD confined to women with low folate and riboflavin intake? Analysis of food records from the Danish osteoporosis prevention study. AB - We have previously found BMD and fracture risk to be significantly associated with the MTHFR (C677T) polymorphism in healthy postmenopausal women in the first years after menopause. Since then, other cohort studies have suggested that sufficient intake of riboflavin and/or folate may have the potential to prevent development of low BMD in women with the TT genotype. This could to some extent explain why this polymorphism is associated with low BMD or fracture in some study populations and not in others. It would also indicate that fractures associated with the TT genotype could be preventable by vitamin B supplementation. We have, therefore, reviewed baseline food record data from our original study to determine if BMD and fracture associations with the MTHFR genotype depended on the intake of folate, riboflavin, or other members of the vitamin B complex, associated with homocysteine metabolism. We analyzed genotype, BMD, and dietary records from 1700 healthy postmenopausal women who participated in the DOPS study. For the assessment of fracture risk, we used longitudinal observations from 854 women in the control group who remained compliant with their initial allocation of no treatment. Riboflavin intake was significantly correlated with femoral neck (FN) BMD in women with the TT genotype (r = 0.24, P < 0.01). FN and lumbar spine (LS) BMD were only associated with the MTHFR genotype in the lowest quartile of riboflavin intake. At the FN, similar threshold effects were shown for folate, vitamin B12, and vitamin B6. Among these vitamin B complex members, stepwise regression analysis identified riboflavin as the only significant predictor of FN BMD in the TT genotype. In conclusion, we confirm reports that BMD in the MTHFR TT genotype is only significantly reduced in the lowest quartile of riboflavin, B12, B6, and folate intake, at least at the time of menopause. Vitamin B supplementation would only be expected to benefit BMD in about 2% of the population, i.e., those with the TT genotype and low vitamin B intake. PMID- 15777681 TI - Influence of estrogen therapy at conventional and high doses on the degree of mineralization of iliac bone tissue: a quantitative microradiographic analysis in postmenopausal women. AB - The beneficial skeletal effects of menopausal estrogen replacement therapy (HRT) are well documented. The role of secondary mineralization of bone as a determinant of bone quality is now well established in postmenopausal women treated with bisphosphonates or SERMs. The aim of present study was to investigate the effect of conventional and high doses of estrogen on the main parameters reflecting the degree of mineralization of bone (DMB). Bone biopsies were obtained from 20 women with osteopenia or osteoporosis before and after 24 months (18 to 38 months) of conventional HRT, and from 19 women who had received high doses of estradiol (implant 100 mg every 3-6 months for 1.5-20 years). DMB parameters (mean DMB, DMB Freq. Max. and Heterogeneity Index of the individual distributions of DMB) were measured using quantitative microradiography in cortical, cancellous, and total bone and expressed as g mineral/cm(3) bone. Values obtained in women before HRT were lower than those reported in pre- and postmenopausal control women. After conventional HRT, there was an increase in mean DMB (total bone) of 4.4 +/- 1.9% (mean +/- SEM) versus pre-treatment values (4.1 +/- 2.1% in cortical bone, 4.5 +/- 2.3% in cancellous bone); these differences did not reach statistical significance (P = 0.055). Results were similar for DMB Freq. Max. but Heterogeneity Index was not significantly changed. After high dose estradiol therapy, mean DMB (total bone) was 6.9 +/- 1.9% higher than in untreated women (8.6 +/- 2.1% in cortical bone, 6.5 +/- 2.1% in cancellous bone); this difference was statistically significant (P T polymorphism was associated with total and intertrochanteric hip BMD and BMD was lower in the 32% of subjects homozygous for the C allele. This polymorphism codes for a nonsynonymous amino acid change with the T allele coding for valine, while the C allele codes for alanine. The difference in BMD was 3.1% (TT vs. CC) and 2.3% (CT versus CC) for the total hip (P = 0.023), and 3.7% (TT vs. CC) and 2.8% (CT versus CC) for the intertrochanter site (P = 0.012). Haplotype analysis demonstrated 6 haplotypes of frequency greater than 2%. A major haplotype defined by G-C-T alleles in SNPs -5826G > A, 3564C > T and 6007C > T respectively, showed association with high bone mass. No SNP showed association with fracture rates. We conclude that a polymorphism found in the BMP4 gene, affecting amino acid sequence, is associated with hip bone density in postmenopausal women, presumably via regulation of anabolic effects on the skeleton. PMID- 15777684 TI - Proximal femur bone geometry is appropriately adapted to lean mass in overweight children and adolescents. AB - It is unclear if the bones of overweight children are appropriately adapted to increased loads. The objective of this study was to compare bone geometry in 40 overweight (body mass index [BMI] > 85th percentile) and 94 healthy weight (BMI < or = 85th percentile) subjects, ages 4-20 years. Dual energy X-ray absorptiometry (Hologic QDR 2000) scans were analyzed at the femoral shaft (FS) and narrow neck (NN) by the Hip Structure Analysis program. Subperiosteal width, cortical thickness and indices of bone axial and bending strength (bone cross-sectional area [CSA] and section modulus [Z]) were measured from bone mass profiles. Multivariate regression models were used to compare overweight and healthy weight subjects. Z was 11 (95% CI 5, 19) and 13 (7, 20) percent higher at the FS and NN, respectively, in overweight subjects (P < 0.001), adjusted for height, maturation and gender. At the NN, higher Z was due to greater subperiosteal width [4% (2, 7)] and bone CSA [10% (5, 16]) and at the FS, to higher bone CSA [10% (5, 16)] and thicker cortices [9% (3, 15)]. When lean mass was added to the models, bone variables did not differ between overweight and healthy weight subjects (P > 0.22), with the exception of NN subperiosteal width [3% (0, 6), P = 0.04]. Fat mass did not contribute significantly to any model. In summary, proximal femur bone geometric strength in overweight children was appropriately adapted to lean mass and height but greater weight in the form of fat mass did not have an independent effect on bone bending strength. These geometric adaptations are consistent with the mechanostat hypothesis that bone strength adapts primarily to muscle forces, not to static loads represented by body weight. PMID- 15777685 TI - Bisphosphonate (YM529) delays the repair of cortical bone defect after drill-hole injury by reducing terminal differentiation of osteoblasts in the mouse femur. AB - We evaluated the effects of YM529, a nitrogen-containing bisphosphonate, on the repair of cortical bone after drill-hole injury at the tissue-, cell- and gene levels in the femur of mice. Eight-week-old male C57BL/6N mice were treated with an intravenous injection of 0.01, 0.05 and 0.1 mg/kg body weight (BW) of YM529, or the vehicle (VC) once a week from 8 weeks of age until sacrifice. At 10 weeks of age (day 0), a drill-hole was made in the diaphysis of bilateral femurs. Femoral specimens were obtained at days 3, 5, 7, 10, 14, 21 and 28 after surgery. Histology and histomorphometry confirmed the early woven bone formation in 7 days after injury in all four groups, but the following lamellar bone repair in the cortical tissue area delayed only in the YM529-treated groups. Since the findings were not dose-dependent, following evaluations were performed in VC and YM529 0.1 mg/kg BW dose groups. Calcein-labeled surface of regenerated bone decreased at day 21 in the YM529 group. At day 0, CFU-f number and mineralized nodule area that developed from marrow cells were significantly smaller in YM529 group than in VC group. At day 5, however, these values increased to levels similar to those in VC group. The mRNA expression levels of BMP-2, cbfa1, osterix, type I collagen, and osteocalcin in the injured bone and marrow cells at days 3 and 5 were similar in the two groups, but were higher in YM529 group at day 7 compared with that in the VC group. At day 14, the levels of these mRNAs were still high, while that of osteocalcin was significantly reduced compared to the VC group. These data indicate that the action of YM529 on bone formation is bimodal, stimulatory on the developments of osteogenic cells for the woven bone regeneration and inhibitory on the terminal differentiation of osteoblasts for the later remodeling, consequently leading to a delay in the lamellar bone healing in the cortical tissue area. PMID- 15777686 TI - Prenatal ethanol exposure has differential effects on fetal growth and skeletal ossification. AB - There is increasing evidence suggesting that the intrauterine environment may influence long-term bone health and the risk of developing osteoporosis in later life. Alcohol (ethanol) is one factor whose presence in the prenatal environment has long-term consequences for the offspring, including permanent growth retardation. Moreover, prenatal ethanol exposure retards both fetal and postnatal bone development. It is unknown if ethanol's effects on skeletal development result from generalized growth retardation or effects specific to skeletal development. Furthermore, the level of ethanol exposure required to produce skeletal effects is unknown. The objectives of this study were to determine (1) if ethanol exerts specific effects on fetal skeletal development that are independent from its effects on general growth, and (2) the level of prenatal ethanol exposure required to affect fetal growth and skeletal ossification. Rats were fed isocaloric diets with ethanol (15%, 25%, or 36% ethanol-derived calories (EDC), approximating low, moderate, and high exposure levels), or without ethanol (pair-fed, PF, or control, C groups), prior to and throughout 21 days of gestation. The degree of E-induced delay in development was determined by comparison of E fetuses on d21 gestation to C fetuses on d17-d21 gestation. Prenatal ethanol exposure at 36% EDC decreased fetal body weight, length, and skeletal ossification compared with PF and C fetuses on d21 gestation. Importantly, effects on ossification, but not body weight or length, were also seen at the more moderate dose of 25% EDC, and the number of bones affected and the severity of effects on ossification tended to increase with dose of ethanol. Comparison of E fetuses on d21 gestation with C fetuses from d17 to 21 gestation indicated that the ethanol-induced delay in development differed for weight and skeletal ossification, and was not uniform among skeletal sites. Taken together, these data suggest that prenatal ethanol exposure has effects on fetal skeletal development that are independent of those on overall fetal growth, and that these effects occur even at moderate levels of maternal drinking. Effects of prenatal ethanol exposure on fetal skeletal development could potentially increase the offspring's risk of osteoporosis later in life. PMID- 15777687 TI - Seroprevalence of Toxoplasma gondii in farm-reared ostriches and wild game species from Zimbabwe. AB - One hundred and seventy one serum samples from 10 game species from Zimbabwe were tested for IgG antibodies to Toxoplasma gondii infection using the modified agglutination test (MAT). Significantly higher seroprevalences were found in the felidae (Panthera leo) (92% of 26), bovidae (Tragelaphus species) (55.9% of 34) and farm-reared struthionidae (Struthio camelus) (48% of 50) compared to the other groups tested. Among the bovidae, the nyala (Tragelaphus angasii) had the highest seroprevalence of 90% (9/10). Anti-Toxoplasma antibody prevalences in browsers [greater kudu (Tragelaphus strepsiceros) (20% of 10), giraffe (Giraffa camelopardalis) (10% of 10) and elephant (Loxodonta africana) (10% of 20)] were generally in the lower range. No antibodies were detected in the wild African suidae [warthog (Phacochoerus africanus) and bushpig (Potamochoerus larvatus)]. Attempts to isolate T. gondii from the heart muscles of seropositve ostriches by subinoculation in BALB/c mice were unsuccessful. PMID- 15777688 TI - Isolation and characterization of a novel PHGPx gene in Raphanus sativus. AB - A full-length cDNA encoding putative phospholipid hydroperoxide glutathione peroxidase (PHGPx) was cloned from Raphanus sativus. The cDNA, designated RsPHGPx, includes an open reading frame which encodes 197 amino acid residues. The alignment of amino acid sequences showed that RsPHGPx had the highest sequence homology to plant PHGPx and contained an N-terminal extension characteristic of a mitochondrial targeting peptide. Northern blot analysis indicated that RsPHGPx was constitutively and ubiquitously expressed during radish development, and its expression was differently regulated by various stress conditions. The expression of RsPHGPx in a yeast PHGPx-deletion mutant significantly rescued the mutant sensitivity to oxidation-sensitive linolenic acid, just as the yeast PHGPx3 gene did. This suggested that RsPHGPx encodes a functional PHGPx protein. PMID- 15777689 TI - Time and frequency domain analyses of heart rate variability in patients with epilepsy. AB - Heart rate variability (HRV) is a useful tool for the detection of sympathetic parasympathetic balance of autonomic nervous system in patients at risk of sudden death (SD). SD is more common in patients with epilepsy and the exact mechanisms of SD are unknown. Autonomic nervous system involvement in patients with epilepsy has rarely been studied and has shown conflicting results. Our purpose was to determine if HRV showed any changes in patients with epilepsy in comparison with normal population. A short period analysis of HRV was performed for both the frequency and time domain in 43 epilepsy patients who had generalized tonic clonic seizures (GTCS) and who were not taking any medications and also in 43 age and sex matched controls. In the time domain analysis, patients displayed higher SDNN (standard deviation of all R-R intervals), SDANN (standard deviation of mean NN intervals in 5 min recordings) and HRV triangular index than did healthy subjects (p < 0.0001). Patients tended to display higher pNN50 (number of R-R intervals differed by > 50 ms from adjacent interval divided by the total number of all R-R intervals) and RMSSD (root-mean-square of successive differences) values than did healthy subjects, but the differences were not statistically significant (p > 0.05). In the frequency domain analysis, the spectral measures of HRV showed a reduction of high frequency (HF) values (is a marker of parasympathetic activity) and an increase of low frequency (LF) values (is a measure of sympathetic activity); as a result, the ratio between low and high frequencies (LF/HF) was significantly increased (p < 0.0001, p < 0.0001 and p < 0.001, respectively). Our data suggests an increase in the sympathetic control of the heart rate in epilepsy patients who have GTCS. This increased sympathetic activity could play a key role in the development of ventricular tachyarrhythmias in patients with epilepsy and may be related to the higher incidence of sudden death in this disorder as compared to controls. PMID- 15777690 TI - The effect of melatonin treatment on oxidative and nitrosative stress in rats with thioacetamide-induced hepatic damage. AB - BACKGROUND: : The following study aimed to clarify the importance of arginase and NOS activities in thioacetamide-induced hepatic damage and to evaluate the underlying mechanism of proposed protection provided by melatonin, using commonly applied therapeutic dose. METHODS: : Rats were randomly assigned to four groups (n=5): control, melatonin (10mg/kg i.p.), thioacetamide (200mg/kg i.p., two doses with a 24h interval) and thioacetamide+three doses of melatonin (10mg/kg i.p., prior- and post-treatment with a 24h interval before thioacetamide administrations) treated groups. RESULTS: : Thioacetamide administration caused hepatic damage creating oxidative and nitrosative stress accompanying perivenous necrosis and eosinophil infiltration. The significant elevation of total nitrite level in livers of thioacetamide treated groups reflected the activation of inducible nitric oxide synthase activity. The decrease in arginase activity indicated hepatic damage. Non-altered specific activity of arginase in the livers of thioacetamide treated groups did not overcome the elevation of NO production. Melatonin treatment did not modulate the levels/activities significantly. CONCLUSIONS: : Our results have indicated that nitrosative stress seems to be essentially critical in thioacetamide-induced hepatic failure in rats. Possible regulatory effect of arginase on NO production and applied dose of melatonin could not prevent hepatic damage. PMID- 15777691 TI - Tribendimidine: a promising, safe and broad-spectrum anthelmintic agent from China. AB - We review, for the first time, a 20-year Chinese story of research and development pertaining to tribendimidine, a promising anthelmintic agent that is safe and exhibits a broad spectrum of activity. Tribendimidine was first synthesized at the National Institute of Parasitic Diseases in Shanghai in the mid 1980s. In laboratory studies, tribendimidine showed high efficacy against Nippostrongylus braziliensis in rats, Necator americanus in hamsters, Ancylostoma caninum and Toxocara canis in dogs, and Syphacia mesocriceti in mice. Activity was also found against several species of cestodes in chicken. In clinical trials, a single oral dose of 400 mg tribendimidine, administered to patients infected only with N. americanus, or with N. americanus and Ancylostoma duodenalis, resulted in cure rates of 85.7% (132/154) and 89.8% (53/59), respectively. In comparison, a single oral dose of 400 mg albendazole resulted in significantly lower cure rates, namely 65.5% (91/139; chi(2) = 16.47, P < 0.001) and 71.7% (43/60; chi(2) = 6.29, P = 0.012), respectively. Single oral doses of tribendimidine (300 mg) and albendazole (400mg) were equally effective against Ascaris lumbricoides infections; cure rates were 96.0% (97/101) and 98.1% (101/103), respectively. In 5-14-year-old children with an Enterobius vermicularis infection, treated with a single oral dose of 200 mg tribendimidine, a cure rate of 81.6% (93/114) was observed. Tribendimidine was well-tolerated as only mild and transient side effects were observed. It would be of great public health significance if these findings are confirmed in other epidemiological settings, as more than one-quarter of the world population is currently affected by intestinal nematodes, with only very few drugs currently available on the market. PMID- 15777693 TI - Evaluation of hepatic functional reserve by scintigraphy with (99m)Tc-GSA. PMID- 15777692 TI - RNA interference of PPARgamma using fiber-modified adenovirus vector efficiently suppresses preadipocyte-to-adipocyte differentiation in 3T3-L1 cells. AB - The peroxisome proliferator-activated receptor (PPAR) gamma is regarded as a "master regulator" of adipocyte differentiation and is abundantly expressed in adipose. To understand the biological role of PPARgamma in adipose, RNA interference (RNAi) of PPARgamma should be a powerful tool. 3T3-L1 cell line serves an excellent model to investigate the mechanism of preadipocyte-to adipocyte differentiation. However, this cell line is difficult to transfect by plasmid vectors and viral vectors. We optimized the transduction of both 3T3-L1 preadipocytes and adipocytes by means of fiber-modified adenovirus (Ad) vectors. Among the various vectors tested, polylysine modification of the C-terminal of the fiber knob most markedly improved the transduction efficiency in both 3T3-L1 preadipocytes and adipocytes. Then, we examined whether fiber-modified Ad vectors with polylysine peptides expressing the small interfering RNA (siRNA) for PPARgamma inhibit the differentiation of 3T3-L1 preadipocytes into adipocytes. Oil red O staining and measurement of glycerol-3-phosphate dehydrogenase (GPDH) activity indicated that the vectors effectively suppressed the differentiation of 3T3-L1 preadipocytes to adipocytes. These results suggested that the combination of fiber-modified Ad vectors containing polylysine peptides and RNAi is an effective tool for the study of the biological and physiological mechanism of adipogenesis in adiposity and diabetes using 3T3-L1 models. Ad vector-mediated RNAi for PPARgamma should also be useful to clarify the biological role of the PPARgamma pathway in various tissues in addition to adipose and for therapeutic application to a variety of diseases, including adiposity and diabetes. PMID- 15777695 TI - Characterization of a novel Eph receptor tyrosine kinase, EphA10, expressed in testis. AB - In mammals, 14 members of the Eph receptor tyrosine kinase family have been described so far. Here we present a not yet described member of this family denoted EphA10. We report the identification of three putative EphA10 isoforms: one soluble and two transmembrane isoforms. One of the latter isoforms lacked the sterile alpha motif commonly found in Eph receptors. The gene encoding EphA10 is located on chromosome 1p34 and expression studies show that EphA10 mRNA is mainly expressed in testis. Binding studies to ephrin ligands suggests that this receptor belongs to the EphA subclass of Eph receptors binding mainly to ephrin-A ligands. PMID- 15777694 TI - Genetic characterization of Entamoeba dispar isolates in Northeast Brazil. AB - The genetic variability of Entamoeba dispar strains was investigated in 39 positive isolates on a survey of 1783 individuals from two different cities of Northeast Brazil (Recife and Macaparana) using two polymorphic species-specific loci (loci 1-2 and 5-6). A combinatory clustering analysis revealed no geographical correlation and remarkable genetic polymorphism among all the isolates examined. Nevertheless, a comparison of the frequency of eight individual PCR products, shared by both Recife and Macaparana populations, for the two loci, showed that only one product of locus 5-6 was significantly different between the two cities. These results suggested that the Macaparana population is infected by similar strains and that locus 5-6 shows potential in assaying questions related to the molecular epidemiology of this region. PMID- 15777696 TI - Looking at structure, stability, and evolution of proteins through the principal eigenvector of contact matrices and hydrophobicity profiles. AB - We review and further develop an analytical model that describes how thermodynamic constraints on the stability of the native state influence protein evolution in a site-specific manner. To this end, we represent both protein sequences and protein structures as vectors: structures are represented by the principal eigenvector (PE) of the protein contact matrix, a quantity that resembles closely the effective connectivity of each site; sequences are represented through the "interactivity" of each amino acid type, using novel parameters that are correlated with hydropathy scales. These interactivity parameters are more strongly correlated than the other hydropathy scales that we examine with: (1) the change upon mutations of the unfolding free energy of proteins with two-states thermodynamics; (2) genomic properties as the genome size and the genome-wide GC content; (3) the main eigenvectors of the substitution matrices. The evolutionary average of the interactivity vector correlates very strongly with the PE of a protein structure. Using this result, we derive an analytic expression for site-specific distributions of amino acids across protein families in the form of Boltzmann distributions whose "inverse temperature" is a function of the PE component. We show that our predictions are in agreement with site-specific amino acid distributions obtained from the Protein Data Bank, and we determine the mutational model that best fits the observed site-specific amino acid distributions. Interestingly, the optimal model almost minimizes the rate at which deleterious mutations are eliminated by natural selection. PMID- 15777697 TI - Analysis of chitin synthase function in a plant parasitic nematode, Meloidogyne artiellia, using RNAi. AB - A single chitin synthase gene is responsible for chitin production in the eggshells of the plant parasitic nematode Meloidogyne artiellia. In this paper we describe a functional analysis of this gene using RNAi as well as further analysis of two similar genes from the free-living nematode Caenorhabditis elegans. The parasitic life-style of M. artiellia required the development of a novel method for delivery of dsRNA to nematode eggs that may be of utility in other experimental systems. C. elegans chitin synthase genes were silenced by feeding nematodes bacteria expressing appropriate chitin synthase dsRNA from a plasmid vector, while M. artiellia egg masses were soaked in dsRNA solution. The results obtained demonstrated that the synthesis of chitin continues to take place in nematode eggs within the egg sac in the parasitic nematode, and that the removal of this activity affects egg development in both C. elegans and M. artiellia. The method described here provides a new way of investigating gene function in plant parasitic nematodes allowing the validity of potential target genes for novel control methods to be assessed. Furthermore, since intact egg cells within the gelatinous matrix of M. artiellia are permeable to dsRNA, eggs of other nematodes may also be similarly permeable to dsRNA and therefore amenable to use with dsRNAi. PMID- 15777698 TI - Expression and transcriptional regulation of Munc18 isoforms in mast cells. AB - Mast cells are specialized immune cells with a central pathophysiological role in allergic reactions and important roles in pathogen defense. Their main effector response is the exocytic release of preformed inflammatory mediators from secretory granules. Munc18 proteins are essential for exocytic function, so we analyzed the expression of Munc18 transcripts in RBL-2H3 mast cells and mouse bone marrow derived mast cells (BMMC). All three isoforms of Munc18 are expressed in both cell types, but Munc18-2 transcripts are most abundant. The proximal 181 bp region of the Munc18-2 gene promoter is conserved between mice and humans, and shows maximal promoter activity among a series of truncation mutants. Binding sites for Ets, E-box and CREB transcription factors that are known to be important for mast cell development are highly conserved and functionally active. Thus, mast cells upregulate an essential component of their exocytic machinery as they develop morphologic and functional features of the regulated secretory phenotype. PMID- 15777700 TI - A role of mast cells for hepatic fibrosis in primary sclerosing cholangitis. AB - We encountered four patients with overt primary sclerosing cholangitis (PSC) which were histologically classified into stage 2 or 3. We examined the expression of stem cell factor (SCF), a ligand of c-kit, in injured bile ducts by immunohistochemistry, and mast cells were identified by immunohistochemistry using anti-HMCT (human mast cell tryptase) and anti-c-kit antibodies to clarify their relation with portal fibrosis coincident with destroyed bile ducts. SCF was detected in the epithelia of most bile ducts in PSC, and many HMCT- and c-kit positive mast cells were found in portal tracts. Image analysis showed more significant numbers of c-kit-positive mast cells per area of portal tract in PSC than in chronic hepatitis C, and they might increase from stage 2 to 3. c-Kit positive cells infiltrated into the portal tracts with SCF-positive destroyed bile ducts, and c-kit mast cells should be investigated in detail to make a role for portal fibrosis in PSC. PMID- 15777699 TI - Intimate relationship between the genes of two transcriptional coactivators, ADA2a and PIMT, of Drosophila. AB - PIMT, a transcriptional coactivator which interacts with and enhances nuclear receptor coactivator PRIP function, was identified recently in mammalian cells and suggested to function as a link between two major multiprotein complexes anchored by CBP/p300 and PBP. Here we describe that the gene of the Drosophila homologue of PIMT, designated as Dtl, is closely associated and has an overlapping promoter with a gene encoding another transcriptional coactivator, ADA2a, which in turn participates in GCN5 HAT-containing complexes. Ada2a also produces an RNA polII subunit, RPB4, via alternative splicing; consequently, an overlapping regulatory region serves for the production of three proteins, each involved in transcription. By studying expression of reporter gene fusions in tissue culture cells and transgenic animals we have demonstrated that the regulatory regions of Ada2a/Rpb4 and Dtl overlap and the Dtl promoter is partly within the Ada2a/Rpb4 coding region. The shared regulatory region contains a DRE element, binding site of DREF, the protein factor involved in the regulation of a number of genes which play a role in DNA replication and cell proliferation. Despite the perfectly symmetrical DRE, DREF seems to have a more decisive role in Ada2a/Rpb4 transcription than in the transcription of Dtl. PMID- 15777701 TI - Biochemical manifestations of anti-tuberculosis drugs induced hepatotoxicity and the effect of silymarin. AB - In the present study, the biochemical manifestations of liver toxicity caused by co-administration of anti-TB drugs, rifampicin (RIF), isoniazid (INH) and pyrazinamide (PZA), in a sub-chronic mode (12 weeks), were investigated. Significant alterations were revealed in (a) increased levels of alanine aminotrasferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) and a high bilirubin content in serum; (b) elevated lipid peroxidation (LPO), intracellular calcium [Ca(2+)](i) and CYP4502EI activity in liver; and (c) decreased glutathione (GSH) content, glutathione peroxidase (GPx) and catalase activities in liver. Silymarin reversed these abnormal alterations. The biochemical changes were supported by histological observations. PMID- 15777702 TI - A possible mechanism of neonatal intrahepatic cholestasis caused by citrin deficiency. AB - For this study, we investigated why cholestasis develops into neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), and concluded that primary mitochondrial impairment associated with the delayed maturity of bile acid metabolism may contribute to the occurrence of NICCD. PMID- 15777703 TI - Comparison of Plasmodium falciparum growth in sickle cells in low oxygen environment and candle-jar. AB - The first successful in vitro cultivation of Plasmodium falciparum in sickle cells in a gas mixture containing 3% oxygen, 4% carbon dioxide and 93% nitrogen has been reported recently, contradicting earlier claims that the parasite does not multiply continuously in sickle cell trait (HbAS) and sickle cell anemia (HbSS) erythrocytes at low oxygen tension. The present study extends that report by growing three P. falciparum strains in erythrocytes from four different sickle cell trait and four sickle cell anemia donors. Because P. falciparum is known to grow normally in sickle cells when incubated in a candle-jar estimated to contain 15-18% oxygen, we have also compared the growth at 3% oxygen with that in a candle-jar. For convenience, we also refer to the 3% oxygen and the candle-jar as low and high oxygen environment, respectively. The three P. falciparum strains were first grown continuously in low oxygen environment for at least 1 month in erythrocytes from one HbAS carrier. These stock cultures were then used to infect erythrocytes from additional three HbAS carriers and four HbSS patients. Results of the experiments showed that parasite growth and hemozoin production in HbAS erythrocytes in low oxygen environment were comparable to those obtained in the candle-jar. There was growth retardation in HbSS erythrocytes in low oxygen environment, but some of the parasites survived and eventually produced high parasitemia levels. Continuous cultivation of different P. falciparum strains in HbAS erythrocytes is necessary for investigation of possible molecular differences between malaria parasites in sickle cells and those in HbAA erythrocytes. PMID- 15777704 TI - Chagas' disease: TCRBV9 over-representation and sequence oligoclonality in the fine specificity of T lymphocytes in target tissues of damage. AB - Using the same mouse strain and two Trypanosoma cruzi sub-populations (CA-I and RA) it is possible to induce pathology in different target tissues: skeletal muscle (CA-I) or sciatic nerve and spinal cord (RA). On the other hand, T cells are directly involved in tissue injury in a strain-dependent way, resembling the abnormalities of chronic Chagas' disease. In the present work, we examined the TCRBV repertoire and the CDR3 sequence polymorphism of T cells infiltrating spinal cord, sciatic nerve and skeletal muscle in chronically infected mice. The TCRBV9 segment was systematically over-represented in the target tissues for each T. cruzi strain: sciatic nerve and spinal cord in RA and skeletal muscle in CA-I infected mice. The analysis of CDR3 sequence polymorphism in the same tissues showed a high proportion of identical TCRBV9 clones in RA-infected mice: 66.6% of the TCRBV9 clones found in sciatic nerve and spinal cord expressed one out of four major CDR3 rearrangements. Sequence identity was shared among clones from sciatic nerve and spinal cord, tissues that are also damaged by passive transfer of CD8 + TL. Those observations are consistent with an antigen driven T-cell expansion sequestered at the inflammation site and demonstrate -- for the first time -- the presence of an oligoclonal repertoire in the antigen recognition site of over-represented T cells in nervous system tissues in chronic Chagas' disease. PMID- 15777705 TI - Dehydroepiandrosterone negatively regulates the p38 mitogen-activated protein kinase pathway by a novel mitogen-activated protein kinase phosphatase. AB - Dehydroepiandrosterone-sulfate, the sulfated form of dehydroepiandrosterone, is the most abundant steroid in young adults, but gradually declines with aging. In humans, the clinical application of dehydroepiandrosterone targeting some collagen diseases, such as systemic lupus erythematosus, as an adjunctive treatment has been applied in clinical trial. Here, we report that dehydroepiandrosterone may negatively regulate the mitogen-activated protein kinase pathway in humans via a novel dual specificity protein phosphatase, DDSP (dehydroepiandrosterone-enhanced dual specificity protein phosphatase). DDSP is highly homologous to LCPTP/HePTP, a tissue-specific protein tyrosine phosphatase (PTP) which negatively regulates both ERK and p38-mitogen-activated protein kinase, and is transcribed from the PTPN7 locus by alternative splicing. Although previous reports have shown that the mRNA expression of the LCPTP/HePTP gene was inducible by extracellular signals such as T-cell antigen receptor stimulation, reverse transcribed (RT)-PCR experiments using specific sets of primers suggested that the expression of LCPTP/HePTP was constitutive while the actual inducible sequence was that of DDSP. Furthermore DDSP was widely distributed among different types of human tissues and specifically interacted with p38-mitogen activated protein kinase. This inducible negative regulation of the p38-mitogen activated protein kinase-dependent pathway may help to clarify the broad range of dehydroepiandrosterone actions, thereby aiding the development of new preventive or adjunctive applications for human diseases. PMID- 15777706 TI - An Ets element regulates the transcription of the human 2B4 gene in natural killer cells. AB - 2B4 (CD244) acts as an activation receptor on human NK cells, whereas it sends inhibitory signals in murine NK cells. A previous study indicated a prominent role for AP-1 in the transcription of 2B4 gene. To further understand the transcriptional regulation we analyzed the upstream positive regulatory region ( 1151 to -704) of the 2B4 promoter. We have identified an Ets element that regulates the 2B4 gene transcription in an AP1 dependent manner. PMID- 15777707 TI - Isolation of Hox and ParaHox genes in the bivalve Pecten maximus. AB - The Hox cluster genes encode a set of transcription factors that have been shown to control spatial patterning mechanisms in bilaterian organism development. The ParaHox cluster is the evolutionary sister of the Hox cluster. The two are believed to descend from an ancestral ProtoHox cluster of four genes from which the three ParaHox genes (Gsx, Xlox, and Cdx) and the four Hox gene classes are believed to have originated. Although molluscs are among the most successful lophotrochozoan groups, very little work has been devoted to the characteristics of their homeotic genes. Using polymerase chain reaction-based approaches, we isolated 13 different Pecten maximus (Bivalvia: Pteriomorphia) sequences corresponding to all the genes of the four Hox cluster classes and to genes Xlox and Cdx of the ParaHox cluster. Comparison of results with those obtained in other lophotrochozoans seems to confirm the considerable homogeneity of the Hox and ParaHox genes in these taxa both as regards the presence of nearly all the genes of the two clusters and the marked sequence resemblance among orthologous genes. PMID- 15777708 TI - uAUG and uORFs in human and rodent 5'untranslated mRNAs. AB - The control of translation is a fundamental mechanism in the regulation of gene expression. Among the cis-acting elements that play a role in translation regulation are upstream open reading frames (uORFs) and upstream AUG (uAUGs) located in the 5'UTR of mRNAs. We present here a genome-wide analysis of uAUGs and uORFs in a curated set of human and rodent mRNAs. Our study shows that the occurrence of uAUGs is suppressed more strongly than that of uORFs and that in frame uAUGs are more strongly suppressed than out-of-frame uAUGs. A very similar pattern of uAUG/uORF frequency was also observed in mouse mRNAs. The analysis of orthologous 5'UTR sequences revealed a remarkable degree of evolutionary conservation only of those uORFs which acquired some functional activity. Our data suggest that besides leaky scanning and reinitiation, which likely occur with variable and gene-specific efficiency, the ribosome-shunt mechanism, eventually coupled to reinitiation after uORF translation, may be a widespread mode of translation regulation in eukaryotes. PMID- 15777709 TI - Puralpha activates PDGF-A gene transcription via interactions with a G-rich, single-stranded region of the promoter. AB - Transcription of the PDGF-A chain gene is regulated by multiple promoter and silencer elements that are GC-rich and exhibit considerable single-stranded character. In this study, the 42 kDa single-stranded DNA and RNA binding protein, Puralpha, was investigated with respect to its ability to bind and interact functionally with single-stranded DNA elements in the PDGF-A gene. Recombinant GST-Puralpha bound with high affinity and sequence-specificity to the G-rich strands of two such transcriptional control elements, the 5'-S1 nuclease hypersensitive silencer (5'SHS; -1418 to -1388) and the nuclease-hypersensitive element (NHE; -92 to -48). Ethylation interference footprinting localized binding of Puralpha to a region between nucleotides -91 and -77 within the NHE element, which contains binding sites for the double-stranded DNA-binding transcription factors Sp1, EGR-1 and WT1. Forced expression of Puralpha upregulated transcriptional activity of the PDGF-A promoter but not the 5'SHS silencer in HepG2 cells, demonstrating Puralpha has the potential to activate PDGF-A gene expression. Targeted disruption of the Puralpha gene reduced NHE activity and PDGF-A mRNA expression in mouse embryo fibroblasts, consistent with a physiological role for Puralpha in maintaining optimal transcription of the PDGF A gene. These results indicate Puralpha enhances transcription of the PDGF-A gene through its interactions with single-stranded, G-rich strands in the promoter, perhaps by stabilizing non-B-form DNA conformations. PMID- 15777710 TI - Novel genes, possibly relevant for molecular diagnosis or therapy of human rhabdomyosarcoma, detected by genomic expression profiling. AB - Transcriptional profiles of an alveolar rhabdomyosarcoma (RMS) and of a RMS cell line were reconstructed by a computational and statistical approach. Expression data of 29,963 genes in 11 adult human healthy tissues and in 37 tumour tissues were analysed for comparison. We identified 202 genes differentially expressed in at least one RMS sample, as compared with normal skeletal muscle. Among them, 107 resulted specifically overexpressed in RMS, but in no tumour affecting other tissues. Cluster analysis applied to expression data detected a series of genes presumably co-expressed with genes encoding known tumour markers and/or reportedly involved in genesis or development of rhabdomyosarcoma. This study succeeded in identifying a number of genes, which become candidates for in vitro study, thus facilitating discovery of novel tumour markers or targets for drug therapy. PMID- 15777711 TI - Dual functions of murine gammadelta cells in inflammation and autoimmunity in coxsackievirus B3-induced myocarditis: role of Vgamma1+ and Vgamma4+ cells. AB - Coxsackieviruses are a cause of clinical myocarditis. Both virus replication and host defense mechanisms, including virus-induced autoimmunity, mediate heart injury and cardiac dysfunction. Vgamma4+ cells kill infected cardiocytes and virus-specific CD4+ Th2 cells through Fas-dependent apoptosis and CD1d. The CD4+ Th1 response is necessary for activation of the autoimmune CD8+ T cells, which kill uninfected cardiocytes through perforin-dependent mechanisms. PMID- 15777712 TI - Murine gammadelta T cells in infections: beneficial or deleterious? AB - Although the importance of gammadelta T cells in pathogen-induced immune responses is becoming increasingly apparent, it is not clear that their involvement is always of benefit to the host. Here we review evidence for the protective and damaging roles of gammadelta T cells in infection and discuss how these disparate findings might be resolved by considering the nature and properties of the pathogen, the sites of infection and conditions under which gammadelta T cell responses are initiated, and the involvement of different subsets of gammadelta T cells. PMID- 15777713 TI - Forum in immunology. Introduction. Non-conventional T cells: useful or harmful players in microbial immunity? PMID- 15777714 TI - A novel ancestral splicing mutation in the multidrug resistance protein 2 gene causes Dubin-Johnson syndrome in Ashkenazi Jewish patients. AB - Dubin-Johnson syndrome (DJS) is an inherited disorder characterized by chronic conjugated hyperbilirubinemia due to the absence or dysfunction of the multidrug resistance protein 2 (MRP2). We previously identified two distinct ancestral mutations causing DJS in 22 unrelated Iranian and five unrelated Moroccan Jewish patients, respectively. In this study we identified and characterized the mutation causing DJS in Ashkenazi Jewish patients and assessed a possible founder effect. Sequencing of all 32 exons of the MRP2 gene identified a novel IVS8+4A- >G mutation in three unrelated homozygotes. Haplotype analysis using four intragenic dimorphisms disclosed a founder effect for the mutation. RT-PCR and real time PCR analysis of mRNA from one patient revealed three splice variants all leading to frameshifts and predicting premature termination codons. The main splice variant was a consequence of the use of a cryptic donor splice site inside exon 8. Liver biopsy in one patient revealed complete absence of MRP2 from the canalicular membrane of hepatocytes. In conclusion, our results provide strong evidence that an ancestral IVS8+4A-->G mutation causes DJS in Ashkenazi Jewish patients by abolishing normal splicing of intron 8 leading to aberrantly spliced products that predict truncation of MRP2. PMID- 15777715 TI - Positive and negative regulatory elements in the late lactation protein-A gene promoter from the tammar wallaby (Macropus eugenii). AB - Little is known about the regulation of the marsupial-specific late lactation protein-A (LLP-A) gene, first expressed at mid-lactation in the mammary gland of the tammar wallaby. A genomic clone of LLP-A was sequenced and shown to include seven exons. The LLP-A promoter region of 1969 bp ligated to a secreted alkaline phosphatase (SEAP) gene reporter was co-transfected into CHO-K1 cells with prolactin (PRL) receptor cDNA. Transfected cells cultured with insulin, cortisol and PRL did not secrete SEAP into media. Similarly, this construct was not expressed in the mammary gland of eight lines of transgenic mice. In contrast, when the LLP-A promoter region was reduced to 850 bp, the expression of the SEAP reporter in CHO-K1 cells was constitutive and PRL-independent, despite the presence of two low affinity Stat5 binding sites. The 1969 bp promoter was analyzed using nine serial deletions ligated to the SEAP gene. The expression of these constructs was PRL-independent. Five putative inhibitory elements were identified between -1969 and -1796, -1404 and -1184, -1184 and -992, -992 and 757, and -591 and -425, and a putative enhancer or core transcription element between -425 and-239. These studies indicate that the complex temporal regulation of the LLP-A gene involves elements in its 5'-regulatory region. PMID- 15777716 TI - Molecular cloning and characterization of a novel androgen repressible gene expressed in the prostate epithelium. AB - Prostate cancer deaths are due to functional escape of prostate cancer cells from their original androgen-dependent growth. To better understand the origin and evolution of hormone-refractory prostate cancer, it is important to identify and characterize genes expressed in the androgen-deprived prostate. We have verified that the rudimentary prostate of congenital androgen deficient mice (hpg) is indeed androgen independent. Using suppression subtractive hybridization between mRNA derived from prostates of hypogonadal (hpg) with or without 14 days of testosterone replacement we have cloned a novel gene from the hpg prostate, termed ADMP (for androgen down regulated gene expressed in mouse prostate), that is down regulated by androgens. ADMP expression is strong in hpg mouse prostate, weak in mature castrated mouse prostate and absent in normal intact or androgen replaced hpg mouse prostates. While ADMP expression is androgen independent in the hpg prostate, it appears to be androgen-dependent in the kidney and brain of normal intact mouse suggesting tissue specific regulation of ADMP by androgens. Human ADMP mRNA expression is suppressed by androgens in the androgen-sensitive LNCaP cell line. The predicted mouse and human protein of 76 amino acids shares sequence similarity to a putative G-protein coupled receptor indicating its possible role in signal transduction. Human ADMP expression was seen predominantly in the prostate epithelium with weaker expression in the fibroblasts and endothelial cells. Cloning and characterization of ADMP has made it feasible to determine its prospective role in the absence of androgens in prostate cancer. PMID- 15777717 TI - Phylogenetic and structural analysis of the Drosophila melanogaster p21-activated kinase DmPAK3. AB - P21-activated kinases (PAKs) are a family of serine/threonine kinases whose diverse cellular functions in cytoskeletal reorganisation, cell motility, transformation and cell death are regulated both by the binding of the small RhoGTPases RAC and CDC42 and by RhoGTPase independent mechanisms. The genome of Drosophila melanogaster encodes three different PAK proteins: DmPAK1, DmMBT (DmPAK2) and DmPAK3. Although structurally related, DmPAK and DmMBT control different developmental processes and also differ in the regulation of their kinase activity through binding of RAC or CDC42. Here, we report the characterisation of DmPAK3. The phylogenetic analysis provides evidence that DmPAK3 and the related proteins from Drosophila pseudoobscura, Anopheles gambiae and Apis mellifera make up a distinct subgroup within the PAK protein family, which might be confined to insects. The structural differences of this PAK subgroup are also reflected by the selective binding of DmPAK3 to RAC-like RhoGTPases. Our biochemical analysis supports a model in which DmPAK3 can form homodimers where the N-terminal regulatory domain of one DmPAK3 protein can bind to and thereby inhibit the catalytic domain of the other DmPAK3 protein. Binding of activated RAC to the regulatory domain or mutation of the RAC-binding site in DmPAK3 relieves the inhibitory effect on kinase activity. Furthermore, our data indicate a function of DmPAK3 in reorganisation of the actin cytoskeleton. PMID- 15777718 TI - Effects of methylation of non-CpG sequence in the promoter region on the expression of human synaptotagmin XI (syt11). AB - We have studied the effects of methylation of the promoter region on the expression of human synaptotagmin XI (syt11), a gene implicated in the onset of schizophrenia. Sequence analysis showed that cytosine residues not in the CpG sequence, but still within the promoter region of the gene, are partially methylated. The methylated cytosine residues are located in the mRNA-coding (minus) strand of the promoter region (mCmCTTmCTTmCmC). Gel mobility shift assays showed that when the cytosine residues are methylated, the binding activity of an Sp family protein, a transcription factor, to the region is significantly reduced. Furthermore, transient transcription assays using artificially methylated promoter sequences showed that methylation did reduce the expression of the reporter gene. The biological significance of the finding is discussed in respect to the effect of methylation of non-CpG sequences in promoter regions on gene expression. PMID- 15777719 TI - Gene expression in the salivary complexes from Haementeria depressa leech through the generation of expressed sequence tags. AB - A survey of the transcriptional profile of Haementeria depressa Ringuelet, 1972 (Annelida, Hirudinea) salivary complexes was produced through expressed sequence tag (EST). Sequences from 898 independent clones were assembled in 555 clusters, representing the transcript profile of this tissue. The repertoire of possible proteins involved in feeding and host interaction processes of the leech corresponded to 10.6% of all identified transcripts (67 clusters), being the carbonic anhydrases (30%), several coagulation inhibitors (25%) and hemerythrin like molecules (19%), the major components. Among the 387 clusters matching cellular proteins, the majority represents molecules involved in gene and protein expression, reflecting a high specialization of this tissue for protein synthesis. Our H. depressa dbEST was also compared to those from other blood feeding organisms, providing evidences that among the secreted proteins, the coagulation inhibitors present a profile very characteristic of this animal class. PMID- 15777720 TI - Polymerase chain reaction (PCR) is highly sensitive for diagnosis of mucosal leishmaniasis. AB - We evaluated the use of polymerase chain reaction (PCR) for diagnosis of mucosal leishmaniasis (ML) in an endemic area in Acre, Brazil, where Leishmania braziliensis is present. Leishmania DNA was detected 34 of 35 cases, yielding a positivity rate of 97.1%, which was higher than the positivity rates for all of the other diagnostic methods studied, namely Montenegro skin test (MST), anti Leishmania serological testing and microscopic examination of lesion biopsy specimens. These findings have led us to propose guidelines for the diagnosis of ML that use PCR as the principal method of parasitological confirmation of cases. PMID- 15777721 TI - Sp-family of transcription factors regulates human SHIP2 gene expression. AB - We have characterized the regulation of human SH-2 containing inositol 5' phosphatase 2 (SHIP2) gene expression. First, the transcription initiation sites and the sequence of the 5' upstream region of human SHIP2 gene were elucidated. Next, the minimal promoter of the human SHIP2 gene was identified by reporter gene assays in HL60 cells and differentiated human subcutaneous white adipocytes. An Sp1 element proximal to the transcription initiation site was indispensable for full promoter activity and bound specifically by Sp1 and Sp3 proteins. These findings suggest that human SHIP2 gene expression, like other housekeeping genes, is controlled by the Sp-family of transcription factors. PMID- 15777722 TI - Characterisation of two novel fork-head gene homologues of Schizosaccharomyces pombe: their involvement in cell cycle and sexual differentiation. AB - The fork-head type transcription factors are a class of regulators that function in a broad spectrum of cellular and developmental processes in many species ranging from yeasts to human. Previous data on yeast fork-head genes suggested roles for these regulators in the control of cell division, sexual differentiation and development. The genome of Schizosaccharomyces pombe has four genes that code for proteins containing fork-head domains (FKH), two of which have been characterised. Here we describe the remaining two genes, fhl1 and fkh2, that code for proteins containing fork-head-associated domains (FHA) besides their FKHs. Neither of them is essential for viability, although the deletion of either fhl1 (putative homologue of Saccharomyces cerevisiae FHL1) or fkh2 (similar to FKH1 and FKH2 of S. cerevisiae) reduced the growth rate and caused an extension of cell length due to delayed G2-to-M transition. Occasionally, multiseptate cells were also produced, indicating the involvement of fhl1 and fkh2 in efficient septum cleavage. The fkh2Delta cells were slightly more sensitive than the wild-type cells to certain environmental stresses, showed reduced fertility and occasional deficiencies in meiosis II, indicating that fkh2 might also act in stress response and sexual differentiation. PMID- 15777723 TI - Molecular characterization and chromosomal assignment of the bovine glycinamide ribonucleotide formyltransferase (GART) gene on cattle chromosome 1q12.1-q12.2. AB - The mammalian glycinamide ribonucleotide formyltransferase (GART) genes encode a trifunctional polypeptide involved in the de novo purine biosynthesis. We isolated a bacterial artificial chromosome (BAC) clone containing the bovine GART gene and determined the complete DNA sequence of the BAC clone. Cloning and characterization of the bovine GART gene revealed that the bovine gene consists of 23 exons spanning approximately 27 kb. RT-PCR amplification of bovine GART in different organs showed the expression of two GART transcripts in cattle similar to human and mouse. The GART transcripts encode two proteins of 1010 and 433 amino acids, respectively. Eleven single nucleotide polymorphisms (SNPs) were detected in a mutation scan of 24 unrelated animals of three different cattle breeds, including one SNP that affects the amino acid sequence of GART. The chromosomal localization of the gene was determined by fluorescence in situ hybridization. Comparative genome analysis between cattle, human and mouse indicates that the chromosomal location of the bovine GART gene is in agreement with a previously published mapping report. PMID- 15777724 TI - Re-analysis of a human hepatitis B virus (HBV) isolate from an East African wild born Pan troglodytes schweinfurthii: evidence for interspecies recombination between HBV infecting chimpanzee and human. AB - According to current estimates, hepatitis B virus (HBV) has infected 2 billion people worldwide and among them, 360 million suffer from chronic HBV infection. Except humans, HBV or HBV-like viruses have also been isolated from different species of apes and mammals. Although recombination has been described to occur extensively between different genotypes within the human HBV lineage, no recombination event has ever been reported between human and non-human primate HBV sequences. It was our objective to perform an exhaustive search for recombination between human and non-human primate HBV strains among all available full-length human and non-human primate HBV sequences, using bootscanning and phylogenetic analyses. Intriguingly, we found that an HBV sequence isolated from a wild born Pan troglodytes schweinfurthii in East Africa-FG-is a recombinant consisting of HBV infecting chimpanzee (ChHBV) and human genotype C. More specifically, in a fragment of approximately 500 nt (positions 551-1050 spanning half of the RT domain of pol, which overlaps with half of the coding region of the small surface protein), FG grouped with HBV genotype C, while in the rest of the genome it grouped with ChHBV sequences. Phylogenetic analyses showed that in the latter region FG was more closely related to the Pan troglodytes troglodytes subspecies, forming an outlier to this group. Moreover, we show evidence that the recombination event occurred after the initial dispersion of HBV genotype C in humans. Finally, our findings point out that although rare recombination between HBV viruses infecting different species occurs. PMID- 15777725 TI - Molecular cloning and characterization of two novel truncated isoforms of human Na+/Ca2+ exchanger 3, expressed in fetal brain. AB - The human gene encoding the Na+/Ca2+ exchanger family member 3 (NCX3) undergoes extensive alternative splicing, with four variants previously identified. In this study, we report two novel alternative transcripts encoding two N-terminally truncated NCX3 proteins specifically expressed in human fetal brain. The identified transcripts, designated NCX3-tN.1 and NCX3-tN.2, are approximately 2.8 kb and 2.9 kb, respectively. The open reading frames (ORFs) are predicted to encode separately a 284 and a 298 amino acid (aa) polypeptide. Sequence analysis and bioinformatics reveal that NCX3-tN.1 and NCX3-tN.2 are the result of alternative splicing of the NCX3 gene. They have their own potential start codons and unique 5' untranslated regions (UTRs) that are different from those of the known NCX3 variants. The variants include a part of intron 2 of the original gene organization as their first exon (exon "a") at the 5' end of the novel transcripts. NCX3-tN.2 consists of six exons including exon "a" and exons 4, 6, 7, 8 and 9 of NCX3, while NCX3-tN.1 lacks exon 4, but is otherwise similar to NCX3-tN.2. Expression studies show that both variants can be translated into protein and NCX3-tN.1 seems more efficiently translated. Based on their structural features, NCX3-tN.1 and NCX3-tN.2 proteins are potentially involved in regulation of Na+/Ca2+ homeostasis. PMID- 15777726 TI - The prion protein gene: identifying regulatory signals using marsupial sequence. AB - The function of the prion protein gene (PRNP) and its normal product PrP(C) is elusive. We used comparative genomics as a strategy to understand the normal function of PRNP. As the reliability of comparisons increases with the number of species and increased evolutionary distance, we isolated and sequenced a 66.5 kb BAC containing the PRNP gene from a distantly related mammal, the model Australian marsupial Macropus eugenii (tammar wallaby). Marsupials are separated from eutherians such as human and mouse by roughly 180 million years of independent evolution. We found that tammar PRNP, like human PRNP, has two exons. Prion proteins encoded by the tammar wallaby and a distantly related marsupial, Monodelphis domestica (Brazilian opossum) PRNP contain proximal PrP repeats with a distinct, marsupial-specific composition and a variable number. Comparisons of tammar wallaby PRNP with PRNPs from human, mouse, bovine and ovine allowed us to identify non-coding gene regions conserved across the marsupial-eutherian evolutionary distance, which are candidates for regulatory regions. In the PRNP 3' UTR we found a conserved signal for nuclear-specific polyadenylation and the putative cytoplasmic polyadenylation element (CPE), indicating that post transcriptional control of PRNP mRNA activity is important. Phylogenetic footprinting revealed conserved potential binding sites for the MZF-1 transcription factor in both upstream promoter and intron/intron 1, and for the MEF2, MyT1, Oct-1 and NFAT transcription factors in the intron(s). The presence of a conserved NFAT-binding site and CPE indicates involvement of PrP(C) in signal transduction and synaptic plasticity. PMID- 15777727 TI - Alternative oxidase and plastoquinol terminal oxidase in marine prokaryotes of the Sargasso Sea. AB - Alternative oxidase (AOX) represents a non-energy conserving branch in mitochondrial electron transport while plastoquinol terminal oxidase (PTOX) represents a potential branch in photosynthetic electron transport. Using a metagenomics dataset, we have uncovered numerous and diverse AOX and PTOX genes from the Sargasso Sea. Sequence similarity, synteny and phylogenetic analyses indicate that the large majority of these genes are from prokaryotes. AOX appears to be widely distributed among marine Eubacteria while PTOX is widespread among strains of cyanobacteria closely related to the high-light adapted Prochlorococcus marinus MED4, as well as Synechococcus. The wide distribution of AOX and PTOX in marine prokaryotes may have important implications for productivity in the world's oceans. PMID- 15777728 TI - Sequence analysis of the Lactobacillus plantarum bacteriophage PhiJL-1. AB - The complete genomic sequence of a Lactobacillus plantarum virulent phage PhiJL-1 was determined. The phage possesses a linear, double-stranded, DNA genome consisting of 36,677 bp with a G+C content of 39.36%. A total of 52 possible open reading frames (ORFs) were identified. According to N-terminal amino acid sequencing and bioinformatic analyses, proven or putative functions were assigned to 21 ORFs (41%), including 5 structural protein genes. The PhiJL-1 genome shows functionally related genes clustered together in a genome structure composed of modules for DNA replication, DNA packaging, head and tail morphogenesis, and lysis. This type of modular genomic organization was similar to several other phages infecting lactic acid bacteria. The structural gene maps revealed that the order of the head and tail genes is highly conserved among the genomes of several Siphoviridae phages, allowing the assignment of probable functions to certain uncharacterized ORFs from phage PhiJL-1 and other Siphoviridae phages. PMID- 15777729 TI - Characterization of multiple lineages of Tc1-like elements within the genome of the amphibian Xenopus tropicalis. AB - We have used genomic sequencing data extracted from the first assembly of the Xenopus tropicalis genome combined with a degenerated PCR approach to identify multiple lineages of Tc1 related transposable elements. Full-length elements were isolated in each lineage and were characterized. Most of them exhibit the typical characteristics of Tc1-like elements (TLEs). An open reading frame (ORF) encoding a 340-350 aa transposase containing a [D, D(34)E] signature was found as well as conserved inverted terminal repeats (ITRs) at each extremities. These ITRs could vary in length, depending on the TLE lineage. These new TLEs were named Eagle, Froggy, Jumpy, Maya, Xeminos, XtTXr and XtTXz. Phylogenetic analyses indicate that their closest relatives are present in the genomes of actinopterygian and amphibian. Interestingly, Maya and Xeminos share remarkable characteristics. Maya contains a [D,D(36)E] motif but is not related to any described TLE so far. Xeminos is the first vertebrate TLE strongly related to an invertebrate lineage. Finally, we have identified for most of these TLEs, copies containing an intact transposase ORF suggesting that these elements may still be active. PMID- 15777730 TI - The role of group 1 and group 2 CD1-restricted T cells in microbial immunity. AB - Group 1 and group 2 CD1 present both self and microbial lipid antigens to T cells. While group 1 CD1-restricted T cells are known for their ability to recognize mycobacterial glycolipid antigens, group 2 CD1-restricted T cells are recognized as regulatory T cells that can influence the outcome of innate and adaptive immune responses. The evidence that these T cells contribute to host defense against infectious diseases is reviewed. PMID- 15777731 TI - Transcription of brain creatine kinase in U87-MG glioblastoma is modulated by factor AP2. AB - Our previous studies established in U87-MG glioblastoma cells that elevated cAMP increased transcription of the endogenous as well as a transiently-transfected brain creatine kinase (CKB) gene, despite the absence of a cAMP response element (CRE) in the CKB proximal promoter. This report employed transfection to show that the transcription of CKB in U87 cells is induced by transcription factor AP2alpha, which is known to be activated by cAMP. Dominant-negative forms of AP2alpha not only prevented the AP2alpha-mediated activation of CKB but also blocked the cAMP-mediated increase in CKB transcription caused by forskolin treatment. The mutation of the four potential AP2 elements within the CKB proximal promoter showed that induction of CKB by AP2 was mediated principally through the AP2 element located at -50 bp in the promoter. Electromobility shift assays revealed a protein in U87 nuclear extracts that bound to a consensus AP2alpha element as well as to the (-50) AP2 element in CKB. Interestingly, the CKB (-50) AP2 element contains GCCAATGGG which also bound NF-Y, the CCAAT-binding protein, suggesting that interplay between AP2 and NF-Y may modulate CKB transcription. This is the first report of a role for AP2 in the regulation of CKB transcription and of an AP2 element within which an NF-Y site is located. PMID- 15777732 TI - Lamotrigine, carbamazepine and phenytoin differentially alter extracellular levels of 5-hydroxytryptamine, dopamine and amino acids. AB - We have studied the effects of treatment with the anticonvulsants lamotrigine (LTG), phenytoin (PHN) and carbamazepine (CBZ) on basal and stimulated extracellular aspartate (ASP), glutamate (GLU), taurine (TAU), GABA, 5 hydroxytryptamine (5-HT) and dopamine (DA) in the hippocampus of freely moving rats using microdialysis. All of the drugs investigated have had inhibition of Na(+) channel activity implicated as their principal mechanism of action. Neither LTG (10-20 mg/kg), PHN (20-40 mg/kg) or CBZ (10-20 mg/kg) had an effect on the basal extracellular concentrations of any of the amino acids studied with the exception of glutamate, which was decreased at the highest LTG dose. However, when amino acid transmitter levels were increased with 50 microM veratridine, LTG was found to cause a dose-dependent decrease in dialysate levels of all four amino acids, with the effect being most pronounced for glutamate. In contrast, PHN decreased extracellular aspartate levels but had no effect on evoked extracellular GLU, TAU or GABA. Somewhat unexpectedly, CBZ did not alter the stimulated increase in the excitatory amino acids, GLU and ASP, but, rather surprisingly for an antiepileptic drug, markedly decreased that of the inhibitory substances TAU and GABA. The three drugs had differing effects on basal extracellular 5-HT and DA. LTG caused a dose-dependent decrease in both, while CBZ and PHN both increased extracellular 5-HT and DA. When extracellular 5-HT and DA was evoked by veratridine LTG had no significant effect on this, while PHN but not CBZ increased stimulated extracellular 5-HT and both PHN and CBZ augmented DA. Thus, the effects of the three drugs studied seemed to depend on whether extracellular transmitter levels are evoked or basal and the particular transmitter in question. This suggests that there are marked differences in the neurochemical mechanisms of antiepileptic drug action of the three compounds studied. PMID- 15777733 TI - Model of transcriptional regulation of the BRCA1-NBR2 bi-directional transcriptional unit. AB - In contrast to hundreds of mutations found in familial breast and/or ovarian cancers, somatic mutations of BRCA1 are very rare. However, a high percentage of sporadic breast and ovarian cancers show a reduction in BRCA1 expression, suggesting that defects in transcriptional regulation is a contributing factor. BRCA1 shares a promoter with its neighboring gene, NBR2, which is transcribed in the opposite direction. We have previously shown that the transcription of BRCA1 is negatively regulated by protein factors that interact with a 36-bp segment, located 575 bp into its first intron. We now report the localization of an 18-bp transcriptional repressor element for NBR2, which resides 948 bp into its first intron. The binding of nuclear proteins to this repressor element was detected by electrophoretic mobility shift assays (EMSAs), and it conferred an orientation dependent functional suppression onto a heterologous thymidine kinase promoter. Combined with our previous studies, a model of transcriptional regulation of the closely aligned BRCA1-NBR2 bi-directional unit is proposed. A minimal 56-bp DNA region is functional in driving transcription in both directions, while uni directional control is provided by distinct repressors that bind to sequences located in the first intron of the respective genes. PMID- 15777734 TI - Hepatic uptake of bilirubin re-visited. PMID- 15777735 TI - Adult-type hypolactasia and regulation of lactase expression. AB - A common genetically determined polymorphism in the human population leads to two distinct phenotypes in adults, lactase persistence and adult-type hypolactasia (lactase non-persistence). All healthy newborn children express high levels of lactase and are able to digest large quantities of lactose, the main carbohydrate in milk. Individuals with adult-type hypolactasia lose their lactase expression before adulthood and consequently often become lactose intolerant with associated digestive problems (e.g. diarrhoea). In contrast, lactase persistent individuals have a lifelong lactase expression and are able to digest lactose as adults. Lactase persistence can be regarded as the mutant phenotype since other mammals down-regulate their lactase expression after weaning (the postweaning decline). This phenomenon does not occur in lactase persistent individuals. The regulation of lactase expression is mainly transcriptional and it is well established that adult-type hypolactasia is inherited in an autosomal recessive manner, whereas persistence is dominant. The recent findings of single nucleotide polymorphisms associated with lactase persistence have made it possible to study the potential mechanisms underlying adult-type hypolactasia. This work has led to the identification of gene-regulatory sequences located far from the lactase gene (LCT). The present review describes the recent advances in the understanding of the regulation of lactase expression and the possible mechanisms behind adult type hypolactasia. PMID- 15777736 TI - Mobility of integrin alpha5beta1 measured on the isolated ventral membranes of human skin fibroblasts. AB - We have measured the lateral mobility of individual alpha5 integrin molecules in ventral plasma membranes of fibroblasts, which were prepared by removal of apical surfaces and nuclei followed by elimination of actin filaments with gelsolin, an actin-severing protein. The cytoplasmic domain of individual integrin molecules was tagged with 100 nm fluorescent polystyrene bead, and motion of the bead was observed and video-recorded. Position of the bead in each frame was determined from the centroid of the fluorescence image, from which plots of the mean-square displacement against time intervals were derived. Within short intervals of time (<100 ms) the mean-square displacement was proportional to the time interval, and the averaged translational diffusion coefficient of (5.3+/-4.4) x 10(-10) cm2/s was obtained with a broad distribution of (1.3-20) x 10(-10) cm2/s. The broad distribution might reflect the oligomerized state of integrin. The largest diffusion coefficient was comparable to that of lipid molecules previously measured in cells and probably represented the diffusion of a single integrin molecule in the presence of little interference of actin cytoskeleton or extracellular matrix. In longer time intervals (>100 ms) the motion of the bead was confined in an area, the average diameter of which was 410+/-160 nm. This was similar to the values described in previous reports, in which the motion of other membrane receptors labeled on their extracellular domain was measured in living cells. PMID- 15777737 TI - Regulatory roles and molecular signaling of TNF family members in osteoclasts. AB - The tumor necrosis factor (TNF) family has been one of the most intensively studied families of proteins in the past two decades. The TNF family constitutes 19 members that mediate diverse biological functions in a variety of cellular systems. The TNF family members regulate cellular functions through binding to membrane-bound receptors belonging to the TNF receptor (TNFR) family. Members of the TNFR family lack intrinsic kinase activity and thus they initiate signaling by interacting intracellular signaling molecules such as TNFR associated factor (TRAF), TNFR associated death domain (TRADD) and Fas-associated death domain (FADD). In bone metabolism, it has been shown that numerous TNF family members including receptor activator of nuclear factor kappaB ligand (RANKL), TNF-alpha, Fas ligand (FasL) and TNF-related apoptosis-inducing ligand (TRAIL) play pivotal roles in the differentiation, function, survival and/or apoptosis of osteoclasts, the principal bone-resorbing cells. These TNF family members not only regulate physiological bone remodeling but they are also implicated in the pathogenesis of various bone diseases such as osteoporosis and bone loss in inflammatory conditions. This review will focus on our current understanding of the regulatory roles and molecular signaling of these TNF family members in osteoclasts. PMID- 15777738 TI - The first characterisation of the overall variability of repetitive units in a species reveals unexpected features of satellite DNA. AB - We investigated the overall variability of the S1a satellite DNA repeats in ten European populations of Rana temporaria by a new procedure that determines the average sequence of the repeats in a genome. The average genomic sequences show that only 17% of the S1a repeat sequence (494 bp) is variable. The variable positions contain the same major and minor bases in all or many of the population samples tested, but the percentages of these bases can greatly vary among populations. This indicates the presence in the species of an enormous number of repeats having a different distribution of bases in these variable positions. Individual genomes contain thousands of repeat variants, but these mixtures have very similar characteristics in all populations because they present the same type of restricted and species-specific variability. Southern blots analyses and sequences of cloned S1a repeats fully support this conclusion. The S1 satellite DNA of other European brown frog species also presents properties indicating the same type of variability. This first characterisation of the overall repeat variability of a satellite DNA in a species has revealed features that cannot be determined by gene conversion and crossing over. Our results suggest that a specific directional process based on rolling circle amplification should play a relevant role in the evolution of satellite DNA. PMID- 15777739 TI - PCR-based approach to SINE isolation: simple and complex SINEs. AB - Highly repeated copies of short interspersed elements (SINEs) occur in eukaryotic genomes. The distribution of each SINE family is usually restricted to some genera, families, or orders. SINEs have an RNA polymerase III internal promoter, which is composed of boxes A and B. Here we propose a method for isolation of novel SINE families based on genomic DNA PCR with oligonucleotide identical to box A as a primer. Cloning of the size-heterogeneous PCR-products and sequencing of their terminal regions allow determination of SINE structure. Using this approach, two novel SINE families, Rhin-1 and Das-1, from the genomes of great horseshoe bat (Rhinolophus ferrumequinum) and nine-banded armadillo (Dasypus novemcinctus), respectively, were isolated and studied. The distribution of Rhin 1 is restricted to two of six bat families tested. Copies of this SINE are characterized by frequent internal insertions and significant length (200-270 bp). Das-1 being only 90 bp in length is one of the shortest SINEs known. Most of Das-1 nucleotide sequences demonstrate significant similarity to alanine tRNA which appears to be an evolutionary progenitor of this SINE. Together with three other known SINEs (ID, Vic-1, and CYN), Das-1 constitutes a group of simple SINEs. Interestingly, three SINE families of this group are alanine tRNA-derived. Most probably, this tRNA gave rise to short and simple but successful SINEs several times during mammalian evolution. PMID- 15777740 TI - DNA microarray analysis of the heat- and cold-shock stimulons in Yersinia pestis. AB - DNA microarray was used as a tool to define the heat- and cold-shock stimulons in Yersinia pestis. Heat shock dramatically enhanced the transcription of genes encoding major heat-shock proteins (MHSPs) that are important for cell survival against the heat. Many other genes were also greatly up-regulated, but their roles in heat-shock response need to be elucidated. Meanwhile, heat shock retarded most of the metabolic processes, i.e. RNA transcription, protein translation, aerobic respiration, energy metabolism, small molecule metabolism, peptidoglycan biosynthesis, sulfate uptake and cysteine biosynthesis. In response to cold shock, Y. pestis has evolved complex adaptive mechanisms by elevating the transcription of a specific set of genes whose protein products are designed to prevent or eliminate cold-induced DNA or RNA structuring, to remodel cell membrane components for maintenance of normal functions, to elevate the energy generation for ensuring ATP-dependent responses during cold adaptation and to synthesize or transport compatible solutes such as cryoprotectants, and at the same time, by repressing the mRNA level of certain genes whose protein products are not needed for bacterial growth at low temperatures, such as the MHSPs. These results provide a set of new candidate genes for hypothesis-based investigations of their roles in stress response, host adaptation and pathogenicity of this deadly pathogen. PMID- 15777741 TI - Mucosal-associated invariant T (MAIT) cells: an evolutionarily conserved T cell subset. AB - Besides mainstream TCRalphabeta T cells harboring a very diverse repertoire, two subsets display an evolutionarily conserved invariant repertoire. This striking conservation indicates important and unique functions. CD1d-restricted NK-T cells expressing an invariant Valpha14 TCRalpha chain have been implicated in microbial and tumor responses as well as in auto-immunity. In this review, we describe the other subset, which bears the canonical hValpha7.2/mValpha19-Jalpha33 TCRalpha chain paired with a restricted set of Vbeta segments. These invariant T cells are present in mice, humans and cattle. They are preferentially located in the gut lamina propria (LP) of humans and mice and are therefore called mucosal associated invariant T (MAIT) cells. Selection/expansion of this population requires B lymphocytes expressing MR1, a monomorphic major histocompatibility complex class I-related molecule that is also strikingly conserved in diverse mammalian species. MAIT cells are not present in germ-free mice, indicating that commensal flora is required for their expansion in the gut LP. The nature of the ligand and the putative functions of these MAIT cells are discussed. PMID- 15777742 TI - The pathogenesis and control of Staphylococcus aureus-induced mastitis: study models in the mouse. AB - The intramammary colonisation by Staphylococcus aureus provokes mastitis in the cow. Once established, the infection is difficult to eradicate with available therapies and may become chronic. The present article focuses on the use of the experimental mouse model of S. aureus-induced mastitis as a practical approach for the study of bovine mastitis. Results obtained regarding the pathogenesis of S. aureus and the development of new therapeutic approaches are discussed. PMID- 15777743 TI - UDP-glucuronosyltransferase1A1 directly binds to albumin. AB - UDP-glucuronosyltransferase1A1 (UGT1A1) catalyses glucuronidation of bilirubin (the final break down product of heme which is produced mainly in the spleen and liver) and is located on the lumen of the endoplasmic reticulum (ER). To identify partner UGTs that form hetero-oligomers with UGT1A1, or other proteins that bind directly to UGT1A1, yeast two-hybrid screening was performed using UGT1A1 as bait. From these studies, cDNA clones specific for human serum albumin (HSA) were unexpectedly isolated. The direct interaction between UGT1A1 and albumin was confirmed in vitro by a pull-down assay. FITC-albumin uptake into HepG2 and Huh7 cells was observed only when bilirubin are present in the culture medium. Furthermore, the endocytosis inhibitor phenylarsine oxide (PAO) prevented albumin uptake into the cells, suggesting that the albumin/bilirubin complex is internalized through receptor-mediated endocytosis. From these studies, it would appear that production of large amounts of toxic bilirubin might use different uptake pathways for entry into hepatocytes. PMID- 15777744 TI - Gene array analysis of Wnt-regulated genes in C3H10T1/2 cells. AB - Wnt/beta-catenin signaling is involved in a large variety of modeling and remodeling processes including cell polarity, cell differentiation, and cell migration. Recently, a role of the Wnt pathway in bone biology has been demonstrated. However, the precise mechanism by which Wnt proteins regulate bone formation still remains to be elucidated. We have previously shown that the Wnt pathway mediates induction of alkaline phosphatase, an osteoblast differentiation marker, in the pluripotent mesenchymal cells C3H10T1/2. In the present study, we performed a genome-wide expression analysis using Affymetrix oligonucleotide chips to determine the Wnt3a-induced gene expression profile in C3H10T1/2 cells. The expression profiles of 447 Wnt3a-regulated genes, classified into distinct functional families, are presented here. Our data reveal that Wnt3a regulates several genes that are involved in osteoblast and adipocyte differentiation. Importantly, Wnt3a induces the expression of osteoprotegerin by a beta-catenin dependent mechanism indicating that the Wnt pathway may also affect osteoclastogenesis. Through the analysis of our expression profiling data, we have established a TaqMan panel as a tool to rapidly compare the expression profiles of a specific set of genes induced by distinct stimuli acting in the Wnt/beta-catenin pathway. Using the TaqMan panel, we have compared the gene expression profiles induced by Wnt1, Wnt2, and Wnt3a in C3H10T1/2 cells, and also by two different GSK-3beta inhibitors: LiCl and SB216773. Our data show that Wnt1 and Wnt3a act in a similar manner, distinct from Wnt2. Finally, we found that LiCl and SB216773 displayed different profiles in the TaqMan panel evidencing their distinct inhibitory action toward GSK-3beta. Overall, data presented herein will aid further understanding of the involvement of the Wnt signaling pathway in its regulation of osteoblast and adipocyte differentiation and function and, in addition, will enhance current knowledge of the Wnt signaling pathway itself. PMID- 15777745 TI - LRP5 gene polymorphisms predict bone mass and incident fractures in elderly Australian women. AB - Postmenopausal osteoporosis and bone mass are influenced by multiple factors including genetic variation. The importance of LDL receptor-related protein 5 (LRP5) for the regulation of bone mass has recently been established, where loss of function mutations is followed by severe osteoporosis and gain of function is related to increased bone mass. The aim of this study was to evaluate the role of polymorphisms in the LRP5 gene in regulating bone mass and influencing prospective fracture frequency in a well-described, large cohort of normal, ambulatory Australian women. A total of 1301 women were genotyped for seven different single nucleotide polymorphisms (SNPs) within the LRP5 gene of which five were potentially informative. The effects of these gene polymorphisms on calcaneal quantitative ultrasound measurements (QUS), osteodensitometry of the hip and bone-related biochemistry was examined. One SNP located in exon 15 was found to be associated with fracture rate and bone mineral density. Homozygosity for the less frequent allele of c.3357 A > G was associated with significant reduction in bone mass at most femoral sites. The subjects with the GG genotype, compared to the AA/AG genotypes showed a significant reduction in BUA and total hip, femoral neck and trochanter BMD (1.5% P = 0.032; 2.7% P = 0.047; 3.6% P = 0.008; 3.1% P = 0.050, respectively). In the 5-year follow-up period, 227 subjects experienced a total of 290 radiologically confirmed fractures. The incident fracture rate was significantly increased in subjects homozygous for the GG polymorphism (RR of fracture = 1.61, 95% CI [1.06-2.45], P = 0.027). After adjusting for total hip BMD, the fracture rate was still increased (RR = 1.67 [1.02-2.78], P = 0.045), indicating factors other than bone mass are of importance for bone strength. In conclusion, genetic variation in LRP5 seems to be of importance for regulation of bone mass and osteoporotic fractures. PMID- 15777746 TI - Simvastatin prevents 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced striatal dopamine depletion and protein tyrosine nitration in mice. AB - Parkinson's disease is a neurological disorder involving the progressive loss of dopaminergic neurons in the substantia nigra pars compacta. There is increasing evidence that inflammation plays a role in the propagation of neurodegenerative processes in Parkinson's disease. We investigated the neuroprotective effects of simvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A inhibitor with anti inflammatory properties, in mice treated with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). Oral administration of simvastatin attenuated the depletion of dopamine, 3,4-dihydroxyphenylacetic acid, and homovanillic acid in the striatum caused by MPTP in a dose-dependent manner. Simvastatin also inhibited the formation of 3-nitrotyrosine in striatal proteins in MPTP-treated mice. Simvastatin had no effect on cholesterol concentrations in the plasma or in the striatum. Simvastatin inhibited the production of tumor necrosis factor (TNF) alpha, nitric oxide, and superoxide in cultured rat microglia stimulated by lipopolysaccharide. The results suggest that simvastatin inhibits the formation of TNF-alpha and peroxynitrite in activated microglia thereby protecting dopaminergic neurons from inflammatory damage. Simvastatin may be a potential new treatment to slow the progression of Parkinson's disease. PMID- 15777747 TI - Acquisition of ischemic tolerance by repetitive transcranial magnetic stimulation in the rat hippocampus. AB - We investigated the acquisition of ischemic tolerance in the rat hippocampus using repetitive transcranial magnetic stimulation (rTMS). Rats received 1000 pulses/day for 7 days, and the field excitatory postsynaptic potentials were measured in the hippocampal CA1. After slices were exposed to ischemic conditions, long-term potentiation (LTP) was induced. The LTP of the stimulated group was enhanced compared with the LTP of the sham control group in each ischemic condition, suggesting that rTMS has the potential to protect hippocampal function from ischemia. PMID- 15777748 TI - The role of impaired insulin/IGF action in primary diabetic encephalopathy. AB - We have previously shown that hippocampal neuronal apoptosis accompanied by impaired cognitive functions occurs in type 1 diabetic BB/Wor rats. To differentiate the contribution by insulin deficiency vs. that by hyperglycemia on neuronal apoptosis, we examined the activities of various apoptotic pathways in hippocampi from type 1 diabetic BB/Wor rats (hyperglycemic and insulinopenic) and type 2 diabetic BBZDR/Wor rats (hyperglycemic and hyperinsulinemic). DNA fragmentation was demonstrated by LM-PCR in type 1 diabetic BB/Wor rats, but was not detectable in duration- and hyperglycemia-matched type 2 BBZDR/Wor rats. Of various apoptotic pathways, Fas activations, 8-OHdG expression, and caspase-12 were demonstrated in type 1 diabetic BB/Wor rats only. In contrast, perturbations of the IGF and NGF systems and PARP activation were demonstrated in type 1 and to a lesser extent in type 2 diabetes. Expressions of Bax and active caspase-3 were significantly increased in type 1, but not in type 2, diabetic rats. These data suggest a lesser apoptogenic stress in type 2 vs. type 1 diabetes. These differences translated into a more profound neuronal loss in the hippocampus of type 1 rats. The results demonstrate that caspase-dependent apoptotic activities dominate in type 1 diabetes, whereas PARP-mediated caspase-independent apoptotic stress is present in both type 1 and type 2 diabetes. The findings suggest that insulin deficiency plays a compounding role to that of hyperglycemia in neuronal apoptosis underpinning primary diabetic encephalopathy. PMID- 15777749 TI - nNOS-containing neurons in the hypothalamus and medulla project to the RVLM. AB - Nitric oxide (NO) within the brain is known to have an important influence on sympathetic nerve activity (SNA). NO is found in the paraventricular nucleus (PVN), caudal ventrolateral medulla (CVLM) and the nucleus tractus solitarius (NTS), regions that project to the rostral ventrolateral medulla (RVLM), an area that is critical in the regulation of SNA. The aim of the present study was to determine whether neurons in the PVN, NTS and CVLM that project to the RVLM contain the neuronal isoform of nitric oxide synthase (nNOS) and are, therefore, capable of producing NO. Under pentobarbitone general anaesthesia, the retrogradely-transported tracer, rhodamine-tagged microspheres, were microinjected into the RVLM of rats (n = 6). Two weeks later, the animals were re anaesthetised, perfused with para-formaldehyde and the brains were removed. Hypothalamic and medullary sections were processed for nNOS immunohistochemistry and the RVLM-projecting neurons were identified using fluorescence microscopy. We found nNOS-containing neurons were present throughout the PVN, CVLM and NTS and that these were intermingled with neurons that projected to the RVLM. Of the neurons in the PVN and CVLM that projected to the RVLM, approximately 12 +/- 1% and 8 +/- 3%, respectively, contained nNOS. In the NTS only 1 +/- 1% of the neurons were double-labeled. This study highlights anatomical pathways emanating from the PVN and CVLM, in particular, which may contribute to the effects on SNA elicited by NO within the brain. PMID- 15777750 TI - Relative sparing of primary auditory cortex in Williams Syndrome. AB - Williams Syndrome (WS) is a neurodevelopment disorder associated with a hemizygous deletion on chromosome 7. WS is characterized with mental retardation, severe visual-spatial deficits, relative language preservation, and excellent facial recognition. Distinctive auditory features include musical ability, heightened sound sensitivity, and specific patterns of auditory evoked potentials. These features have led to the hypothesis that the dorsal forebrain is more affected than the ventral. Previously, we reported primary visual area 17 abnormalities in rostral striate cortex, a region contributing to the dorsal visual pathway. Based on the dorsal-ventral hypothesis, and language and auditory findings, we predicted a more normal histometric picture in auditory area 41. We used an optical dissector method to measure neurons in layers II-VI of area 41 in right and left hemispheres of the same 3 WS and 3 control brains used in the area 17 study. There was a hemisphere by diagnosis interaction in cell packing density (CPD) in layer IV and in cell size in layer III between WS and control brains. Post hoc analysis disclosed in control brains, but not WS, a layer IV left > right asymmetry in CPD, and a layer III left < right asymmetry in cell size. WS brains showed more large neurons bilaterally in layer II and in left layer VI. Histometric alterations in area 41 were less widespread than rostral visual cortex. Also, there was less asymmetry in the WS brain. We interpret layers II and VI differences as reflecting increased limbic connectivity in primary auditory cortex of WS. PMID- 15777751 TI - Extracellular correlates of glutamate toxicity in short-term cerebral ischemia and reperfusion: a direct in vivo comparison between white and gray matter. AB - Glutamate toxicity and cellular calcium overload are thought to be pathophysiological key factors not only in gray matter (GM) but also in white matter (WM) ischemia. Correlates of excitotoxicity have never been directly investigated in vivo in GM and WM ischemia and reperfusion. We measured simultaneously amino acids, purines, and calcium in relation to tissue depolarization using microdialysis and ion-selective electrodes and regional CBF using hydrogen clearance in GM and WM of cats during 10 min of global ischemia and 120 min of reperfusion. CBF ceased during ischemia. Reperfusion was followed by hyperperfusion that turned into hypoperfusion within 60 min in both GM and WM. Direct current potential decreased in ischemia to around -15 mV in GM and -10 mV in WM and shifted back after reperfusion towards control levels in both compartments. Extracellular calcium decreased in GM during ischemia, whereas it increased in WM. After reperfusion, calcium returned to control levels in both GM and WM. Glutamate, aspartate, GABA, and taurine increased in GM but not in WM during ischemia and reperfusion. Adenosine increased transiently in both compartments peaking during the first minutes of reperfusion, and returned thereafter to control levels. Contrasting with GM, deleterious processes such as glutamate accumulation and cellular Ca(2+) influx do not occur in WM during short term ischemia and reperfusion. Rather, an intrinsic neuroprotective role of adenosine may be discussed. In our view, therefore, therapeutic strategies against glutamate toxicity in short-term ischemia and reperfusion should be mainly focused on GM. PMID- 15777752 TI - HMG CoA reductase inhibitors reduce ischemic brain injury of Wistar rats through decreasing oxidative stress on neurons. AB - Statins possess neuroprotective effect against ischemic damage, but how they protect neurons is not exactly made clear. We speculated that anti-oxidative property of statins is implicated, and investigated statins' influences on the oxidative neuronal damage in the brain after ischemia. After 14 days of atorvastatin, pitavastatin, simvastatin, or vehicle administration, 90 min of middle cerebral artery occlusion was imposed on Wistar rats. The production of 4 hydroxynonenal (HNE) and 8-hydroxy-2'-deoxyguanosine (8-OHdG), both of which are oxidative stress markers, as well as infarction formation were investigated at 1 day after the reperfusion. In the vehicle group, massive infarction was confirmed and HNE and 8-OHdG are robustly produced. In the statins-treated group, the infarction was smaller and the HNE and 8-OHdG production was less prominent than the vehicle group. Among the statins investigated, simvastatin was most effective for reducing oxidative stress and infarction volume, which may be brought by its highly lipophilic property. Reduction of oxidative stress by statins may be one main reason in ameliorating ischemic brain damage in rats. PMID- 15777753 TI - Calpain inhibitor MDL 28170 protects hypoxic-ischemic brain injury in neonatal rats by inhibition of both apoptosis and necrosis. AB - MDL 28170 is a CNS-penetrating calpain inhibitor, and we examined the effects of MDL 28170 on hypoxic-ischemic brain injury in immature brain using the Rice Vannucci model. Immediately after hypoxic exposure, 24 mg/kg of MDL 28170 was injected intraperitoneally as an initial dose, followed by 12 mg/kg every 4 h for a total dose of 60 mg/kg over 12 h post-HI. A vehicle control group received peanut oil injection instead. Macroscopic evaluation of brain injury revealed the neuroprotective effect of MDL 28170 after 12 h post-HI. Neuropathological quantitative analysis of cell death showed that MDL 28170 significantly decreased the number of necrotic cells in all the examined regions except for cingular cortex, and the number of apoptotic cells in caudate putamen, parietal cortex, hippocampus CA1, and laterodorsal thalamus. Western blots showed that MDL 28170 suppressed 145/150 kDa subunits of alpha-spectrin breakdown products (SBDP) in cortex, hippocampus, thalamus, and striatum, and also 120-kDa subunit of SBDP in all regions except for striatum. This suggests that MDL 28170 inhibited activation of calpain and caspase-3, respectively. Our results indicate that post hypoxic MDL 28170 injection is neuroprotective in HI newborn rat brain by decreasing both necrosis and apoptosis. SBDP expression also suggests that MDL 28170 injection inhibits both calpain and caspase-3 activation after HI insult. PMID- 15777754 TI - Responses of neuronal nitric oxide synthase expression in the brainstem to electroacupuncture Zusanli (ST 36) in rats. AB - Recent studies have reported that l-arginine-derived nitric oxide (NO) in the gracile nucleus modifies the hypotensive responses to electroacupuncture (EA) stimulation of Zusanli (ST 36). The purpose of this study was to examine the influence of EA stimulation of ST 36 on neuronal NO synthase (nNOS) expression in the brainstem nuclei in rats. EA stimulation of ST 36 and a non-acupoint was performed using 3 Hz of stimulation for 10 s every 2 min for a period of 120 min in rats anesthetized with ketamine. Rats in the sham-treated group received surgery and EA needles were placed into the acupoints without performing the stimulation. After 2-h stimulation and sham treatment, animals were perfused with 4% paraformaldehyde. Sections of rat medulla were examined by immunolabeling with a polyclonal antibody directed against nNOS. The brainstem nuclei were also visualized by NADPH-diaphorase histochemistry, a marker of nNOS activity. nNOS expression and NADPH-diaphorase reactivity were quantified by using a microscope with reticule grid to count the number of positive cells over a nucleus. Unilateral EA stimulation of ST 36 in rats caused increases in nNOS immunostained cells in the rostral region of the ipsilateral gracile nucleus, but was not altered in the contralateral gracile nucleus compared with sham-treated rats (P < 0.05, n = 6-7). NADPH-diaphorase-positive cells were also increased in the ipsilateral gracile nucleus of rats with EA stimulation. nNOS immunostaining and NADPH-diaphorase-positive neurons were significantly increased in both ipsilateral and contralateral sides of the medial nucleus tractus solitarius (mNTS) in rats receiving EA ST 36 compared with sham-treated animals (P < 0.05). nNOS immunostaining and NADPH-diaphorase reactivity was neither altered in the gracile nucleus and mNTS of non-acupoint stimulated rats nor other brainstem nuclei in rats with EA ST 36. These results show that nNOS immunoreactivity and NADPH-diaphorase reactivity are consistently increased in the gracile nucleus and the mNTS by EA ST 36. We conclude that EA ST 36 induces nNOS expression in the gracile nucleus and mNTS, and enhanced nNOS-NO in the nuclei may modify central cardiovascular regulation, which contribute to hypotensive effects of acupuncture. PMID- 15777755 TI - bFGF and EGF modulate trauma-induced proliferation and neurogenesis in juvenile organotypic hippocampal slice cultures. AB - Since postnatal and adult mammalian brains have been shown to retain an ability to generate neurons from endogenous stem cells throughout life, these cells could play a central role in regeneration after neuronal loss. Therefore, we studied cell proliferation, glio- and neurogenesis respectively after brain injury in organotypic hippocampal slice cultures using a focal trauma by transecting Schaffer collaterals in the cornu ammonis (CA) 2 region mechanically. After determination of cell death using propidium iodide, neuroregenerative processes were quantitatively analyzed by various immunohistochemical techniques at different time points post injury. As this endogenous insult-induced neurogenesis is rather inefficient, we investigated if it can be enhanced by application of exogenous growth factors. Exogenous basic fibroblast growth factor (bFGF) enhanced neurogenesis significantly in the dentate gyrus (DG) region. A neutralizing antibody against endogenous bFGF revealed a significant decrease of basal and trauma-induced proliferation. Reverse transcription polymerase chain reaction (RT-PCR) studies exhibited a downregulation of FGF messenger ribonucleic acid (mRNA) transcription after the antibody treatment. In contrast, epidermal growth factor (EGF) increased proliferation, but not neurogenesis. A combination of bFGF and EGF displayed an EGF-like effect on proliferation and no effect on neurogenesis. These results demonstrate, that in our model bFGF but not EGF sustains neurogenesis, whereas together the two growth factors permit an increased proliferation but not neurogenesis in organic hippocampal slice cultures. PMID- 15777756 TI - Mechanisms of 4-hydroxynonenal-induced neuronal microtubule dysfunction. AB - We have previously demonstrated that neuronal microtubules are exquisitely sensitive to the lipid peroxidation product 4-hydroxynonenal (HNE). The mechanism, however, by which HNE disrupts the microtubules, is not known. Sulfhydryl groups of protein-cysteines constitute main targets of HNE. Indeed, HNE is mainly detoxified by conjugation to glutathione (GSH), a reaction that leads to depletion of cellular GSH. GSH maintains protein sulfhydryl groups in the reduced form and has been implicated in the regulation of cytoskeletal function. Here, we assess what role depletion of cellular GSH plays in the HNE induced microtubule disruption. We demonstrate that HNE and its intracellularly activated tri-ester analog, HNE(Ac)(3), cause substantial GSH depletion in Neuro2A cells. However, other compounds inducing GSH depletion had no effect on the microtubule network. Therefore, HNE-induced depletion of cellular GSH does not contribute to the HNE-induced microtubule disruption. We previously demonstrated that another main cellular target of HNE is tubulin, the core protein of microtubules containing abundant cysteines. The functional relevance of this adduction, however, had not been evaluated. Here, we demonstrate that exposure of Neuro 2A cells to HNE or HNE(Ac)(3) results in the inhibition of cytosolic taxol-induced tubulin polymerization. These and our previous observations strongly support the hypothesis that HNE-adduction to tubulin is the primary mechanism involved in the HNE-induced loss of the highly dynamic neuronal microtubule network. PMID- 15777757 TI - Inhibitory influence of ginsenoside Rb(3) on activation of strychnine-sensitive glycine receptors in hippocampal neurons of rat. AB - Whole-cell patch-clamp technique was used to investigate the effect of ginsenoside Rb(3) (Rb(3)), an active constituent of Panax ginseng, on glycine receptor activity in immature hippocampal neurons, which were dissociated acutely from hippocampal CA(1) area in Sprague-Dawley rats aging 10-14 days using the method of enzyme digestion with mechanical dissociation. As a result, glycine elicited an inward current (I(gly)) in a concentration-dependent manner in approximately 86% of those isolated neurons tested. This current was strychnine sensitive. Rb(3) itself did not elicit any membrane currents. However, coapplication of Rb(3) inhibited peak current of I(gly). This depressant effect of Rb(3) varied with its concentrations. At a concentration of 0.1 micromol/L, ginsenoside Rb(3) had the most significant inhibition, with a net reduction of 31% in average. Moreover, the inhibition of I(gly) by Rb(3) did not depend on the membrane potential. Rb(3) (0.1 micromol/L) presented inhibitory effect on I(gly) mainly at higher glycine concentrations (>100 micromol/L), and decreased maximal glycine efficacy. This effect was the same as that of a non-competitive antagonist of glycine receptors. Finally, we found that Rb(3) prolonged the time constant of activation of I(gly). It is therefore suggested that ginsenoside Rb(3), possibly as a non-competitive antagonist, could inhibit strychnine sensitive glycine current at a dose-dependent manner in acutely dissociated hippocampal CA(1) neurons of young rats, and decrease of affinity of glycine to receptors and delay of receptor activation may be involved in this inhibition. Inhibitory effect of ginsenoside Rb(3) on I(gly) is possibly one of the bases of many pharmacological actions of Panax ginseng. PMID- 15777758 TI - Immunocytochemical and stereological analysis of GABA(B) receptor subunit expression in the rat vestibular nucleus following unilateral vestibular deafferentation. AB - The process of behavioral recovery that occurs following damage to one vestibular labyrinth, vestibular compensation, has been attributed in part to a down regulation of GABA(B) receptors in the vestibular nucleus complex (VNC) ipsilateral to the lesion, which could potentially reduce commissural inhibition from the contralateral VNC. In this study, we tested the possibility that this occurs through a decrease in the expression of either the GABA(B1) or GABA(B2) subunits of the GABA(B) receptor. We used Western blotting to quantify the expression of these subunits in the VNC at 10 h and 50 h following unilateral vestibular deafferentation (UVD) or sham surgery in rats. We then used immunocytochemistry and stereological counting methods to estimate the number of neurons expressing these subunits in the MVN at 10 h and 2 weeks following UVD or sham surgery. Compared to sham controls, we found no significant changes in either the expression of the two GABA(B) receptor subunits in the VNC or in the number of MVN neurons expressing these GABA(B) receptor subunits post-UVD. These results suggest that GABA(B) receptor expression does not change substantially in the VNC during the process of vestibular compensation. PMID- 15777759 TI - Sympathoexcitatory effects of estrogen in the insular cortex are mediated by GABA. AB - The current investigation examined the effect of estrogen in the insular cortex (IC) on autonomic tone and cardiac baroreceptor reflex function and sought to determine if modulation of neurotransmission was responsible for mediating this effect. Experiments were performed in Inactin-anaesthetized, male Sprague-Dawley rats. Animals were instrumented to record blood pressure, heart rate, vagal parasympathetic and renal sympathetic nerve activities, as well as cardiac baroreflex sensitivity (BRS). Direct, bilateral injection of 17beta-estradiol (0.5 microM; 200 nl/side) into the IC resulted in a significant increase in sympathetic tone (from 10 +/- 4 to 24 +/- 3) with no significant change in blood pressure, heart rate, parasympathetic tone or BRS measured at 30 min post injection. This estrogen-induced effect was completely blocked by the co injection of estrogen with the estrogen receptor antagonist, ICI 182, 780 (20 microM; 200 nl/side). Co-injection of estrogen with a GABA(B), NMDA or non-NMDA receptor antagonists did not effect the estrogen-induced increase in sympathetic tone. Co-injection of a sub-threshold dose of estradiol (0.125 microM; 200 nl/side) with the GABA(A) receptor antagonist, (+)-bicuculline (0.025 microM; 200 nl/side), resulted in an additive response to increase sympathetic nerve activity. These results suggest that estrogen acts on estrogen receptors to modulate GABA(A)-receptor-mediated neurotransmission within the IC to modulate sympathetic tone. PMID- 15777760 TI - Differential modulation of the glutamate transporters GLT1, GLAST and EAAC1 by docosahexaenoic acid. AB - At present, the ability of polyunsaturated fatty acids (PUFAs) to regulate individual glutamate transporter subtypes is poorly understood and very little information exists on the mechanism(s) by which PUFAs achieve their effects on the transport process. Here we investigate the effect of cis-4,7,10,13,16,19 docosahexaenoic acid (DHA) on the activity of the mammalian glutamate transporter subtypes, GLT1, GLAST and EAAC1 individually expressed in human embryonic kidney (HEK) cells. Exposure of cells to 100 muM DHA increased the rate of d [(3)H]aspartate uptake by over 72% of control in HEK(GLT1) cells, and by 45% of control in HEK(EAAC1) cells. In contrast, exposure of HEK(GLAST) cells to 200 muM DHA resulted in almost 40% inhibition of d-[(3)H]aspartate transport. Removal of extracellular calcium increased the inhibitory potential of DHA in HEK(GLAST) cells. In contrast, in the absence of extracellular calcium, the stimulatory effect of DHA on d-[(3)H]aspartate uptake in HEK(GLT1) and HEK(EAAC1) cells was abolished, and significant inhibition of the transport process by DHA was observed. Inhibition of CaM kinase II or PKC had no effect on the ability of DHA to inhibit transport into HEK(GLAST) cells but abolished the stimulatory effect of DHA on d-[(3)H]aspartate transport into HEK(GLT1) and HEK(EAAC1) cells. Inhibition of PKA had no effect on the modulation of d-[(3)H]aspartate transport by DHA in any of the cell lines. We conclude that DHA differentially modulates the GLT1, GLAST and EAAC1 glutamate transporter subtypes via different mechanisms. In the case of GLT1 and EAAC1, DHA appears to stimulate d [(3)H]aspartate uptake via a mechanism requiring extracellular calcium and involving CaM kinase II and PKC, but not PKA. In contrast, the inhibitory effect of DHA on GLAST does not require extracellular calcium and does not involve CaM kinase II, PKC or PKA. PMID- 15777761 TI - Hyperbaric oxygen induces rapid protection against focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: The timing and mechanisms of protection by hyperbaric oxygen (HBO) in cerebral ischemia have only been partially elucidated. We monitored the early in vivo effects of HBO after 2 h transient focal ischemia using repetitive MRI. METHODS: Wistar rats underwent filament occlusion of the middle cerebral artery (MCAO). 40 min after MCAO, rats were placed in a HBO chamber and breathed either 100% O(2) at 3.0 atmospheres absolute (ata; n = 24) or at 1.0 ata (control; n = 24) for 1 h. Diffusion, perfusion and T2-weighted MR images were obtained after 15 min and 3, 6 and 24 h of reperfusion. In 6 axial MR slices, volume of abnormal diffusion and T2w signals were measured in the ischemic hemisphere. Furthermore, hemispheric mean apparent diffusion coefficient (ADC) and T2 values were calculated for statistical analysis. RESULTS: HBO significantly reduced volume of abnormal DWI signal beginning immediately after reperfusion (control: 92 +/- 28 mm(3); HBO: 64 +/- 17) and lesion size on T2w (control: 375 +/- 91 mm(3); HBO: 225 +/- 39) after 24 h. Correspondingly, mean ADC levels were lower and T2 values higher in the ischemic hemisphere in the control group. HBO reduced histological infarct size at 24 h. CONCLUSION: High dose intraischemic HBO therapy has an immediate protective on the brain which is superior to normobaric oxygen. PMID- 15777762 TI - Isoflurane and sevoflurane affect cell survival and BCL-2/BAX ratio differently. AB - Depletion of calcium from the neuronal endoplasmic reticulum (ER) induces apoptosis. Isoflurane depletes calcium from sarcoplasmic reticulum (SR) of muscle, an analogue of ER in neurons, while sevoflurane maintains or increases SR calcium. We hypothesized that isoflurane, but not sevoflurane, induces apoptosis by depleting the ER calcium. Rat PC12 pheochromocytoma cells and primary cortical neurons were treated with equipotent doses of isoflurane and sevoflurane. Isoflurane, but not sevoflurane, at equipotent doses induced cell damage determined by both LDH release and MTT reduction assays, dose and time dependently, in both types of cells. Isoflurane at 2.4% for 24 h induced cytotoxicity in both cell types, which was characterized by nuclear condensation and fragmentation and activation of caspases 3 and 9. Isoflurane cytotoxicity was suppressed by dantrolene, a ryanodine receptor antagonist that inhibits abnormal calcium release from the ER. Isoflurane decreased the Bcl-2/Bax ratio by as much as 36% (P < 0.05). However, sevoflurane did not cause neuronal damage by apoptosis nor did it decrease the Bcl-2/Bax ratio. These results suggest that isoflurane and sevoflurane differentially affect the Bcl-2/Bax ratio and cell survival. At equipotent concentrations, isoflurane, but not sevoflurane, induces cytotoxicity in both PC12 cells and primary cortical neurons and decreases the Bcl-2/Bax ratio. PMID- 15777763 TI - Differential activation of trigeminal C or Adelta nociceptors by infrared diode laser in rats: behavioral evidence. AB - Radiant heat is often used for studying thermal nociception, although inherent characteristics such as the broad spectrum of applied wavelengths of typical light sources limit control over and repeatability of stimuli. To overcome these problems, we used a diode infrared laser-based stimulator (wavelength: 980 nm) for selectively stimulating trigeminal Adelta or C thermonociceptors in rats. To provide indirect evidence for nociceptor-selective stimulation, we tested the effects of capsaicin, dimethylsulfoxide (DMSO), and morphine on withdrawal latencies for long pulses with a low current (hypothesized to selectively stimulate C nociceptors) and for threshold currents of short pulses with high current (hypothesized to selectively stimulate Adelta nociceptors) in lightly anesthetized rats. Nonmem analysis was used to perform pharmacodynamic modeling. The measured baseline withdrawal latency for long pulses was 12.5 +/- 0.3 s which was changed significantly to 6.7 +/- 0.4 s after applying topical capsaicin which selectively sensitizes C nociceptors and to 16.5 +/- 1.3 s after 1.0 mg/kg morphine which preferentially attenuates C fiber nociception. Topical DMSO which appears to selectively sensitize Adelta afferents did not significantly alter withdrawal latencies to the long pulses. Fitted threshold currents for short pulses after DMSO were however significantly lower (974 +/- 53 mA vs. 1113 +/- 12 mA for baseline) indicating Adelta sensitization. Capsaicin and morphine did not significantly change threshold currents. Best Nonmem fits for the long pulse were obtained using a model assuming no DMSO effect, but a different inter-individual variability after applying this substance. For the short pulse, a model assuming no capsaicin or morphine effect, but again allowing different inter-individual variabilities after applying these drugs, best described the data. We conclude that different settings of the stimulator used in this study were capable of selectively activating trigeminal Adelta or C thermonociceptors. PMID- 15777764 TI - Learning deficits induced by sleep deprivation and recovery are not associated with altered [(3)H]muscimol and [(3)H]flunitrazepam binding. AB - Several studies have shown that sleep deprivation produces deficits in learning tasks, but mechanisms underlying these effects remain unclear. Other lines of evidence indicate an involvement of brain GABA systems in cognitive processes. Here, we investigated the possibility that alterations in GABA(A) or benzodiazepine (BDZ) receptor binding might underlie avoidance deficits induced by sleep deprivation. Rats were deprived of sleep for 96 h using the platform method and then trained in a step-through inhibitory avoidance task, or allowed to recover sleep for 24 h before training (sleep rebound group). Thirty minutes after training, animals were given a retention test. Both sleep-deprived and sleep-recovered animals showed a significant impairment in avoidance responding compared to cage controls, and the sleep-deprived group performed significant worse than the sleep-recovered group. A separate group of animals was sacrificed either immediately after 96 h of sleep deprivation or after 96 h of sleep deprivation followed by 24 h of sleep recovery. [(3)H]muscimol and [(3)H]flunitrazepam binding were examined by quantitative autoradiography in 42 brain regions, including areas involved in cognitive processes. No significant differences among groups were found in any brain region, except for a reduction in [(3)H]flunitrazepam binding in the frontal cortex of sleep-recovered animals. These results confirm the deleterious effects of sleep loss on inhibitory avoidance learning, but suggest that such deficits cannot be attributed to altered GABA(A) or BDZ binding in brain. PMID- 15777765 TI - Electroacupuncture stimulates hypothalamic aromatization. AB - We have previously reported that the repeated electroacupuncture (EA) stimulation significantly increased the concentrations of circulating estradiol and restored the depressed function of the hypothalamus-pituitary-ovary axis (HPOA) in ovariectomized (OVX) rats. We hypothesize that extragonadal aromatization in specific brain areas might be responsible for these changes. Thus, various assays, including radiometric assay, Western blot, and reverse transcriptase polymerase chain reaction (RT-PCR), were employed to determine the aromatization in the hypothalamus of rats that received both OVX and electroacupuncture (OVX + EA). The results showed that EA significantly increased the aromatase activity as well as the expressions of its mRNA and protein (P < 0.05) in the OVX rats. These results suggest that EA enhances brain aromatization, which might contribute to influence the function of gonadotropin-releasing hormone (GnRH) neurons and promote the hypofunction of the HPOA in the ovariectomized rats. PMID- 15777766 TI - Differential effects of Na-K-ATPase pump inhibition, chemical anoxia, and glycolytic blockade on membrane potential of rat optic nerve. AB - Na(+)-K(+)-ATPase pump failure during either anoxia or ouabain perfusion induces rapid axonal depolarization by dissipating ionic gradients. In this study, we examined the interplay between cation and anion transporting pathways mediating axonal depolarization during anoxia or selective Na(+)-K(+)-ATPase inhibition. Compound resting membrane (V(m)) potential of rat optic nerve was measured in a grease gap at 37 degrees C. Chemical anoxia (2 mM NaCN or NaN(3)) or ouabain (1 mM) caused a loss of resting potential to 42 +/- 11% and 47 +/- 2% of control after 30 min, respectively. Voltage-gated Na(+)-channel blockade was partially effective in abolishing this depolarization. TTX (1 microM) reduced depolarization to 73 +/- 10% (chemical anoxia) and 68 +/- 4% (ouabain) of control. Quaternary amine Na(+) channel blockers QX-314 (1 mM) or prajmaline (100 microM) produced similar results. Residual ionic rundown largely representing co efflux of K(+) and Cl(-) during chemical anoxia in the presence of Na(+)-channel blockade was further spared with DIDS (500 microM), a broad-spectrum anion transport inhibitor (95 +/- 8% of control after 30 min in anoxia + TTX vs. 73 +/- 10% in TTX alone). Addition of DIDS was slightly more effective than TTX alone in ouabain (74 +/- 5% DIDS + TTX vs. 68 +/- 4% in TTX alone, P < 0.05). Additional Na(+)-entry pathways such as the Na-K-Cl cotransporter were examined using bumetanide, which produced a modest albeit significant sparing of V(m) during ouabain-induced depolarization. Although cation-transporting pathways play the more important role in mediating pathological depolarization of central axons, anion-coupled transporters also contribute to a significant, albeit more minor, degree. PMID- 15777767 TI - Modulation of glutamate-induced outward current by prostaglandin E(2) in rat dissociated preoptic neurons. AB - The preoptic/anterior hypothalamus (POA) is one of the major brain regions where cytokines and their related mediators (i.e., prostaglandins) exert diverse actions. In the present study, the modulatory effects of prostaglandin E(2) (PGE(2)) on the glutamate-induced membrane currents were examined using perforated-patch clamp method in rat POA neurons that had been mechanically dissociated by vibration without enzyme treatment. Application of glutamate through U-tube induced a slow outward current following fast inward ionotroic current at a holding membrane potential of -30 mV. The slow outward current was also induced by N-methyl-d-aspartate (NMDA), accompanied by an increased membrane conductance, and inhibited by perfusion with Ca(2+)-free solution, tetraethylammonium chloride (TEA), and apamin, suggesting a Ca(2+)-dependent K(+) current (KCa) activated by Ca(2+) entry through NMDA channels. Perfusion with PGE(2) at 0.1-10 microM, a principal mediator of fever and neuroendocrine control at the POA, did not produce apparent current by itself, but selectively potentiated the glutamate- or NMDA-induced KCa without affecting inward currents. The KCa induced by activation of NMDA receptors may serve as a feedback mechanism and the modulatory effects of PGE(2) on the KCa may have an important physiological significance. PMID- 15777768 TI - Selective modulation of Ca(2+) influx pathways by 5-HT regulates synaptic long term plasticity in the hippocampus. AB - Both long-term potentiation (LTP) and long-term depression (LTD) can be induced in the Schaffer collateral-CA1 synapse of the hippocampus either by repetitive stimulation of afferent fibres with the frequency of the stimulation determining the polarity of the response or by associative pairing of pre- and postsynaptic activity. An increase in postsynaptic intracellular Ca(2+) concentration is an important signal for the induction of long-term synaptic plasticity. In patch clamp experiments on hippocampal brain slices, we tested the modulation of different forms of synaptic plasticity by the neurotransmitter serotonin (5-HT) which is known to inhibit high-voltage activated Ca(2+) channels. 1 microM of 5 HT inhibited homosynaptic LTD induced by low frequency stimulation. This effect of 5-HT could be blocked by the selective 5-HT(1A) antagonist WAY 100635. Low frequency-induced LTD is both dependent on Ca(2+) influx through NMDA receptors and high-voltage activated Ca(2+) channels. It was blocked by the NMDA-receptor antagonist D-AP5 and by the N-type Ca(2+) channel antagonist omega-conotoxin GIVA. Tetanus induced LTP was not affected by low concentrations of 5-HT, whereas depotentiation of LTP by asynchronous pairing of EPSPs and postsynaptic action potentials was completely abolished with 5-HT in the bath solution. We conclude that those forms of plasticity which depend on Ca(2+) influx via high-voltage activated Ca(2+) channels are subject to modulation by 5-HT. This might be a relevant mechanism by which 5-HT modifies basic network properties in the brain. PMID- 15777769 TI - Expression of neurocan after transient middle cerebral artery occlusion in adult rat brain. AB - Neurocan is one of the major chondroitin sulfate proteoglycans in the nervous tissues. The expression and proteolytic cleavage of neurocan are developmentally regulated in the normal rat brain, and the full-length neurocan is detected in juvenile brains but not in normal adult brains. Recently, some studies showed that the full-length neurocan was detectable even in the adult brain when it was exposed to mechanical incision or epileptic stimulation. In the present study, we demonstrated by Western blot analysis that the full-length neurocan transiently appeared in the peri-ischemic region of transient middle cerebral artery occlusion (tMCAO) in adult rat with a peak level at 4 days after tMCAO. Immunohistochemical analysis showed that a clear positive signal of neurocan was observed 4 days after tMCAO in the peri-ischemic region of cerebral cortex and caudate, where cells strongly positive in GFAP expression were also distributed. These results indicate that accumulation of the full-length neurocan produced by reactive astrocytes may be one of the processes for tissue repair and reconstruction of neural networks after focal brain ischemia as well. PMID- 15777770 TI - Xanthine oxidase, nitric oxide synthase and phospholipase A(2) produce reactive oxygen species via mitochondria. AB - The formation of reactive oxygen species (ROS) has been suggested to be associated with excitotoxicity but the involvement of cytoplasmic enzymes in ROS formation is not clearly known. In the present study, we examined the role of xanthine oxidase (XO), nitric oxide synthase (NOS) and phospholipase A(2) (PLA(2)) in glutamate-induced oxidative stress in rat cortical slices. Glutamate induced ROS formation and mitochondrial depolarization were measured in rat cortical slices in presence of allopurinol, L-NAME and 4-bromophenacylbromide, the specific inhibitors of XO, NOS and PLA(2), respectively. Upon stimulation of slices with glutamate, a significant increase in ROS formation and mitochondrial depolarization was observed. However, pretreatment of slices with allopurinol, L NAME and 4-bromophenacylbromide inhibited the glutamate-induced ROS formation and mitochondrial depolarization. The glutamate-induced ROS formation was dependent on the concentration of these inhibitors and also on the duration of the treatment. Allopurinol was found to be less effective as compared to L-NAME and 4 bromophenacylbromide. The combined treatment of slices with these enzyme inhibitors showed further inhibition in ROS formation and mitochondrial depolarization. The inhibition in ROS formation as well as mitochondrial depolarization by allopurinol, L-NAME and 4-bromophenacylbromide clearly suggests that the activation of XO, NOS and PLA(2) by calcium during glutamate receptor stimulation may release some chemicals which depolarize mitochondria resulting in ROS formation. PMID- 15777771 TI - Central administration of IGF-I and BDNF leads to long-lasting antidepressant like effects. AB - Drug development research has identified neurotrophic factors as a downstream target of chronic antidepressant treatments. In order to study their antidepressant-like effects, two neurotrophic factors, brain-derived neurotrophic factor and insulin-like growth factor I, were examined in the rat modified forced swimming test after a single icv administration. Both neurotrophins produced antidepressant-like behavioral effects in the modified rat forced swimming test, reducing immobility and increasing swimming. In contrast to currently used antidepressants, which produce acute effects in the forced swimming test, the effects of the neurotrophins were unusually long lasting and persisted at least 6 days after the treatment. Neither neurotrophic factor had an effect on locomotor activity. The results support a role for neurotrophic factors mediating the behavioral effects of antidepressant drugs. PMID- 15777772 TI - Chronic melatonin therapy fails to alter amyloid burden or oxidative damage in old Tg2576 mice: implications for clinical trials. AB - Melatonin has been proposed as a treatment for Alzheimer's disease based on the demonstration of antioxidant and "anti-amyloid" effects in vitro and in vivo. Chronic melatonin therapy in old, amyloid plaque-bearing transgenic mice was studied. Tg2576 mice started melatonin treatment at 14 months of age. After 4 months of treatment, there were no differences between untreated and melatonin treated mice in cortical levels of soluble, formic acid extracted, or histologically detectable beta amyloid (Abeta), nor in brain levels of lipid peroxidation product (total 8,12-isoprostane F(2alpha)-VI), despite marked elevations in plasma melatonin. We conclude that melatonin fails to produce anti amyloid or antioxidant effects when initiated after the age of amyloid plaque deposition. These findings diminish the possibility that melatonin will be useful for the treatment of established Alzheimer's disease. PMID- 15777773 TI - Effects of propofol on recombinant AMPA receptor channels. AB - The interaction of the anaesthetic propofol with recombinant human AMPA-type glutamate receptor channels was investigated by a patch-clamp study using fast agonist application techniques. Despite the marked effects of propofol on inhibitory synaptic transmission and voltage gated sodium channels, there is also evidence for a specific pharmacological action on AMPA receptors. In our study, we observed a deceleration of AMPA receptor channel desensitization in the prolonged presence of glutamate and propofol that is likely to account for the enhancement of ion currents through AMPA receptor channels observed in previous studies. While there was an increase in the rate and extent of desensitization at glutamate receptor 1, glutamate receptor 2, and glutamate receptor 3 AMPA receptors, no affection of current rise time, peak current amplitude, and deactivation properties was observed. Thus, our findings point to an isolated interaction with processes that control desensitization of AMPA receptor channels rather than indicating an interaction with channel opening and closing processes due to agonist binding and unbinding. The pharmacological effect described resembles in part that of compounds like cyclothiazide and aniracetam which are known to interact with channel desensitization. PMID- 15777775 TI - Role of 5-HT1A and 5-HT1B receptors in the antinociceptive effect of tramadol. AB - Tramadol, (1RS,2RS)-2-[(dimethylamine)-methyl]-1-(3-methoxyphenyl)-cyclohexanol hydrochloride, is an atypical centrally acting analgesic agent with relatively weak opioid receptor affinity and which, like some antidepressants, is able to inhibit the reuptake of serotonin (5-hydroxytryptamine, 5-HT) in the raphe nucleus. We have previously demonstrated that pindolol, a beta-adrenoceptor blocker/5-hydroxytryptamine(1A/1B) receptor antagonist, enhanced tramadol antinociception and that the selective 5-HT1A agonist 8-Hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) reduced it. These effects were related to the negative feedback control that regulates raphe region neurones. The current study examines the ability of the selective antagonist at somatodendritic 5-HT1A receptors, N-[2-[4-(2-methoxyphenyl)-1-piperazinyl] ethyl]-N-(2-pyridinyl) cyclohexane carboxamide (WAY100635, 0.8 mg/kg), the selective antagonist at terminal 5-HT1B receptors, N-[3-(2-dimethylamino) ethoxy-4-methoxyphenyl]-2' methyl-4'-(5-methyl-1,2,4-oxadiazol-3-yl)-(1,1'-biphenyl)-4-carboxamide (SB216641, 0.1-0.8 mg/kg) and the selective agonist at 5-HT1B receptors, 1,4 tDihydro-3-(1,2,3,6-tetrahydro-4-pyridinyl)-5H-pyrrolo[3,2-b] pyridin-5-one (CP93129, 0.2-0.4 mg/kg), to modify the antinociceptive effect of 4-64 mg/kg of tramadol in the hot plate test in mice. The results show that 0.8 mg/kg of WAY100635 enhanced antinociceptive effect of tramadol while neither agonism nor antagonism at the 5-HT1B receptor modifies it significantly at the doses tested. These results account for involvement of the somatodendritic 5-HT1A receptors in the analgesic effect of tramadol and support the supraspinal interaction of serotonin and the opioid system in the regulation of pain. PMID- 15777774 TI - Anxiolytic-like effect of a serotonergic ligand with high affinity for 5-HT1A, 5 HT2A and 5-HT3 receptors. AB - S-(-)-2-[[4-(napht-1-yl)piperazin-1-yl]methyl]-1,4-dioxoperhydropyrrolo[1,2 alpha]-pyrazine (CSP-2503) is a serotonin (5-HT) receptor ligand with selectivity and high affinity for 5-HT1A, 5-HT2A and 5-HT3 receptors. CSP-2503 reduced rectal temperature and 5-HT neuronal hypothalamic activity in mice, decreased electrical activity of raphe nuclei cells in rats and blocked the enhancement of adenylate cyclase activity induced by forskolin in HeLa cells transfected with the human 5 HT1A receptor. This compound also blocked head-twitches induced by the 5 HT(2A/2C) receptor agonist 2,5-dimethoxy-4-iodoamphetamine (DOI). Contractions of guinea pig ileum induced by the 5-HT3 receptor agonist 2-methyl-5-HT were prevented by CSP-2503. Moreover, it reduced the bradycardia reflex induced by 2 methyl-5-HT in anaesthetized rats. In the light/dark box and social interaction tests, CSP-2503 presented anxiolytic activity, an action shared by 5-HT1 agonists and 5-HT3 antagonists. Taken together, these results suggest that CSP-2503 is a new 5-HT1 receptor agonist with 5-HT2A and 5-HT3)receptor antagonist activities that might be useful in a number of conditions associated with anxiety. PMID- 15777776 TI - Morphine causes a delayed increase in glutamate receptor functioning in the nucleus accumbens core. AB - Enhanced excitatory neurotransmission in the mesocorticolimbic system may contribute to the persistence of addiction behaviour. Here, we demonstrated that glutamate-, N-methyl-D-aspartate (NMDA)- and alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid (AMPA)-induced [3H]-gamma-aminobutyric acid (GABA) release from superfused rat nucleus accumbens core slices is profoundly enhanced 3 weeks, but not 3 days, after a single s.c. morphine injection. This delayed increase in glutamate receptor functioning is associated with enhanced gene transcript levels of ionotropic NMDA and AMPA/kainate receptor subunits. These data reveal that morphine may progressively enhance glutamate neurotransmission within the nucleus accumbens core subsequent to drug exposure. PMID- 15777777 TI - Disconnection between the early onset anorectic effects by C75 and hypothalamic fatty acid synthase inhibition in rodents. AB - In order to explore the relationship between the anorectic effect of 3-carboxy-4 octyl-2-methylenebutyrolactone (C75) and its pharmacokinetic properties, studies of in vivo and in vitro pharmacological characterization of C75 were performed in Fischer rats. In a quantitative measurement of food intake, we determined that appetite suppression by C75 takes place within 4 h. The C(max) for C75 of 2.6+/ 1.5 microM was reached within 1-4 h after intraperitoneal administration at 30 mg/kg, a drug level that causes complete blockade of food intake. However, this concentration is substantially lower than the effective concentration used to inhibit rat fatty acid synthase enzyme activity in vitro (IC50: approximately 200 microM) and hypothalamic enzyme activity was found not to be inhibited by intraperitoneal administration of C75 at 30 mg/kg. Instead, a dramatic induction of c-Fos expression was found in area postrema. Collectively, these data indicate that the anorectic effect of C75 is independent of its inhibition of fatty acid synthase in the hypothalamus. PMID- 15777778 TI - Serotonin1A-receptor antagonism blocks psychostimulant properties of diethylpropion in marmosets (Callithrix penicillata). AB - Diethylpropion (1-phenyl-2-diethylamine-1-propanone hydrochloride) is a stimulant drug with reinforcing properties that is used to treat obesity in humans. While the anorectic properties of diethylpropion are mediated by a noradrenergic mechanism, stimulant properties depend on its effects on the serotonergic (5-HT) and/or dopaminergic systems. In this study we investigated the role of the 5-HT1A receptor in the acute behavioral effects of diethylpropion in marmosets (Callithrix penicillata). Animals were pretreated with the selective 5-HT1A receptor antagonist, N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2 pyridinyl) cyclohexane-carboxamide trihydrochloride (WAY 100635; 0.2, 0.4, 0.8 mg/kg, i.p.) or saline (i.p.) and received a treatment with diethylpropion (10 mg/kg, i.p) or saline (i.p.). Diethylpropion induced an increase in locomotor activity in 60% of the monkeys, which were classified as diethylpropion sensitive, but did not affect locomotion in 40% of the monkeys (diethylpropion insensitive). Sensitivity analysis revealed two types of responders to diethylpropion. In the sensitive animals (type A) diethylpropion increased locomotor activity and anxiogenic-like behavior, but decreased bodycare activities. In the insensitive animals (type B) diethylpropion did not affect locomotor and bodycare activity after diethylpropion, but led to a strong increase in anxiogenic-like behavioral responses. Selective 5-HT1A-receptor antagonism modulated the acute diethylpropion effects responder type specifically. In the sensitive (type A) monkeys WAY 100635 blocked the diethylpropion-induced increase in locomotor activity, while not affecting anxiogenic-like behavioral responses or the suppression of bodycare activities. In the insensitive monkeys, WAY 100635 had no effect on locomotor activity after diethylpropion, but blocked diethylpropion effects on some anxiogenic-like behavioral responses. In conclusion, these results suggest an essential contribution of the 5-HT1A-receptor to the stimulant effects of diethylpropion, which is responder type specific. It also suggests the 5-HT1A-receptor to be a source of the interindividual variance in the acute behavioral response to the stimulant diethylpropion in monkeys. PMID- 15777779 TI - Nitric oxide dynamics and endothelial dysfunction in type II model of genetic diabetes. AB - Although diabetes is a major risk factor for vascular diseases, e.g., hypertension and atherosclerosis, mechanisms that underlie the "risky" aspects of diabetes remain obscure. The current study is intended to examine the notion that diabetic endothelial dysfunction stems from a heightened state of oxidative stress induced by an imbalance between vascular production and scavenging of reactive oxygen/nitrogen species. Goto-Kakizaki (GK) rats were used as a genetic animal model for non-obese type II diabetes. Nitric oxide (NO) bioavailability and O2- generation in aortic tissues of GK rats were assessed using the Griess reaction and a lucigenin-chemiluminescence-based technique, respectively. Organ chamber-based isometric tension studies revealed that aortas from GK rats had impaired relaxation responses to acetylcholine whereas a rightward shift in the dose-response curve was noticed in the endothelium-independent vasorelaxation exerted by the NO donor sodium nitroprusside. An enhancement in superoxide (O2-) production and a diminuation in NO bioavailability were evident in aortic tissues of GK diabetic rats. Immunoblotting and high-performance liquid chromatography (HPLC)-based techniques revealed, respectively, that the above inverse relationship between O2- and NO was associated with a marked increase in the protein expression of nitric oxide synthase (eNOS) and a decrease in the level of its cofactor tetrahydrobiopterin (BH4) in diabetic aortas. Endothelial denudation by rubbing or the addition of pharmacological inhibitors of eNOS (e.g. N(omega) nitro-L-arginine methyl ester (L-NAME)), and NAD(P)H oxidase (e.g. diphenyleneiodonium, apocynin) strikingly reduced the diabetes-induced enhancement in vascular O2- production. Aortic contents of key markers of oxidative stress (isoprostane F2alpha III, protein-bound carbonyls, nitrosylated protein) in connection with the protein expression of superoxide generating enzyme NAD(P)H oxidase (e.g. p47phox, pg91phox), a major source of reactive oxygen species in vascular tissue, were elevated as a function of diabetes. In contrast, the process involves in the vascular inactivation of reactive oxygen species exemplified by the activity of CuZnSOD was reduced in this diseased state. Our studies suggest that diabetes produces a cascade of events involving production of reactive oxygen species from the NADPH oxidase leading to oxidation of BH4 and uncoupling of NOS. This promotes the oxidative inactivation of NO with subsequent formation of peroxynitrite. An alteration in the balance of these bioactive radicals in concert with a defect in the antioxidant defense counteracting mechanism may favor a heightened state of oxidative stress. This phenomenon could play a potentially important role in the pathogenesis of diabetic endothelial dysfunction. PMID- 15777780 TI - The ablation of the Ca(v)2.3/E-type voltage-gated Ca2+ channel causes a mild phenotype despite an altered glucose induced glucagon response in isolated islets of Langerhans. AB - Glucagon release upon hypoglycemia is an important homeostatic mechanism utilized by vertebrates to restore blood glucose to normal. Glucagon secretion itself is triggered by Ca2+ influx through voltage-gated ion channels, and the gene inactivation of R-type Ca2+ channels, with Ca(v)2.3 as the ion conducting subunit, has been shown to disturb glucose homeostasis. To understand how glucagon release may be affected in Ca(v)2.3-deficient mice, carbachol, insulin and glucose induced glucagon response was investigated. While the rise of insulin and glucose induced by carbachol is normal, mutant mice show an impaired glucagon response. Further, the effect of insulin injection on glucagon levels was altered by the loss of the Ca(v)2.3 subunit. Ca(v)2.3-deficient mice are characterized by an impaired glucose suppression of glucagon release. This was most obvious at the level of isolated islets suggesting that Ca(v)2.3 containing R-type voltage-gated Ca2+ channels are involved in the glucose-mediated signalling to glucagon release in mice. PMID- 15777781 TI - Formalin-induced pain is reduced in sigma(1) receptor knockout mice. AB - The role of sigma1 receptors in non-acute pain has not been explored. In this study we show that both phases of formalin-induced pain were reduced by approximately 55% in sigma1 receptor knockout mice in comparison to wild-type animals. These results suggest that the tonic pain induced by formalin is altered in mice lacking sigma1 receptors, and highlight the potential usefulness of further studies of the role of sigma1 receptors in models of non-acute pain. PMID- 15777782 TI - Seeing is believing! The optical properties of the eye lens are dependent upon a functional intermediate filament cytoskeleton. AB - Beaded filaments are the major cytoskeletal element of the eye lens and they are essential to the optical properties of the eye lens. They were discovered in 1972 by Harry Maisel and Margaret Perry and have since been found to comprise two novel intermediate filament proteins, CP49 and filensin. These proteins possess unique structure features and unusual assembly characteristics, which distinguish them from canonical IF proteins. Whilst CP49 is completely tailless, filensin has a rather short rod domain and extremely large C-terminal tail domain. In vitro, CP49 and filensin do not form IFs on their own. In vitro studies suggest that CP49 and filensin have a distinct coassembly mechanism. Whilst CP49 self assembles into thick bundles of filaments, filensin only forms short fibrils, but when combined together they form filaments. The generation of gene knockouts by the targeted deletion of Bfsp1 and Bfsp2 that encode filensin and CP49, respectively, have been made to explore the function of beaded filaments in the lens. Our results suggest that the lens-specific beaded filaments are the key cytoskeletal element in organising and maintaining lens fibre cell architecture and are a key factor in determining the optical properties of the lens. We have also found that some common mouse strains contain a natural mutation in Bfsp2 that will effectively generate a CP49 knockout. This finding has important implications for lens research involving other gene knockouts maintained on a 129 background. It has also been observed that mutations in Bfsp2 are the genetic basis of inherited human cataract. Collectively, these data demonstrate that beaded filaments are fundamental to lens function. PMID- 15777783 TI - Binding sequence-dependent regulation of the human proliferating cell nuclear antigen promoter by p53. AB - Exposure of a lung epithelial cell line to ionizing radiation (IR) arrests cell cycle progression through 48 h post-exposure. Coincidentally, IR differentially activates expression of the cell cycle inhibitor, p21/WAF1, and the DNA replication protein, proliferating cell nuclear antigen (PCNA). p21/WAF1 mRNA levels remain elevated through 48 h post-exposure to IR, while PCNA mRNA levels increase transiently at early times. Since p21/WAF1 inhibits DNA replication by directly binding PCNA, the relative levels of the two proteins can determine cell cycle progression. The PCNA p53-binding site displayed reduced p53 binding affinity in vitro relative to the distal p21/WAF1 p53-binding site. Substitution of the p21/WAF1 site for the resident p53-binding site in the PCNA promoter altered the responses to increasing amounts of p53 or IR in transient expression assays. The p21/WAF1 p53-binding site sustained activation of the chimeric PCNA promoter under conditions (high p53 levels or high dose IR) that the PCNA p53 binding site did not. Binding site-specific regulation by wild-type p53 was not observed with mutant p53 harboring a serine to alanine change at amino acid 46. Limited activation of the PCNA promoter by p53 and its modified forms would restrict the amount of PCNA made available for DNA repair. PMID- 15777784 TI - Hepatitis C virus E2 protein promotes human hepatoma cell proliferation through the MAPK/ERK signaling pathway via cellular receptors. AB - Dysregulation of mitogen-activated protein kinase (MAPK) signaling pathways by various viruses has been shown to be responsible for viral pathogenicity. The molecular mechanism by which hepatitis C virus (HCV) infection caused human liver diseases has been investigated on the basis of abnormal intracellular signal events. Current data are very limited involved in transmembrane signal transduction triggered by HCV E2 protein. Here we explored regulation of the MAPK/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway by E2 expressed in Chinese hamster oval cells. In human hepatoma Huh-7 cells, E2 specifically activated the MAPK/ERK pathway including downstream transcription factor ATF-2 and greatly promoted cell proliferation. CD81 and low density lipoprotein receptor (LDLR) on the cell surface mediated binding of E2 to Huh-7 cells. The MAPK/ERK activation and cell proliferation driven by E2 were suppressed by blockage of CD81 as well as LDLR. Furthermore, pretreatment with an upstream kinase MEK1/2 inhibitor U0126 also impaired the MAPK/ERK activation and cell proliferation induced by E2. Our results suggest that the MAPK/ERK signaling pathway triggered by HCV E2 via its receptors maintains survival and growth of target cells. PMID- 15777785 TI - Mesenchymal stem cells inhibit lymphocyte proliferation by mitogens and alloantigens by different mechanisms. AB - Human mesenchymal stem cells (MSCs) have immuno-modulatory properties. They inhibit T-cell proliferation to mitogens and alloantigens in vitro and prolong skin graft survival in vivo. We found that MSCs inhibited the proliferation of peripheral blood lymphocytes (PBLs) to phorbol myristate acetate (PMA), suggesting that MSCs exert an inhibitory effect downstream of the receptor level. We analyzed cytokine profiles of PBLs co-cultured with MSCs. MSCs increased interleukin (IL)-2 and soluble IL-2 receptor in mixed lymphocyte cultures (MLCs), while IL-2 and IL-2R decreased in phytohemagglutinin (PHA)-stimulated PBL cultures. MSCs inhibited IL-2 induced proliferation, without absorbing IL-2. IL 10 levels increased in MLCs co-cultured with 10% MSCs, while the levels were not affected in PHA cultures. In MLCs inhibited by MSCs, antibodies against IL-10 further suppressed proliferation but had no effect in PHA cultures. Addition of indomethacin, an inhibitor of prostaglandin-synthesis, restored part of the inhibition by MSCs in PHA cultures. However, indomethacin did not affect MSC induced inhibition in MLCs. To conclude, our data indicate that MSC-induced suppression is a complex mechanism affecting IL-2 and IL-10 signaling and may function differently, depending on T-cell stimuli. Prostaglandins are important in the inhibition by MSCs when the T cells were activated by PHA, but not alloantigens. PMID- 15777786 TI - Reversible and repeatable linear local cell force response under large stretches. AB - Large stretching and un-stretching force response of adherent fibroblasts is measured by micromachined mechanical force sensors. The force sensors are composed of a probe and flexible beams. The probe, functionalized by fibronectin, is used to contact the cells. The flexible beams are the sensing element. The sensors are made of single crystal silicon and fabricated by the SCREAM process. The maximum cell stretch reached is approximately 50 microm, which is about twice of the cell initial size, and the time delay between two consecutive stretching/un-stretching steps is 75 s unless otherwise stated. We find that the force response of the cells is strongly linear, reversible, and repeatable, with a small stiffening at the initial deformation stage. Force response of single cells measured before and after cytochalasin D treatment suggests that actin filaments take almost all the cell internal forces due to stretch. These findings may shed light on the increasing understanding on the mechanical behavior of cells and provide clues for making new classes of biological materials having uncommon properties. PMID- 15777787 TI - Formation of nucleoplasmic protein aggregates impairs nuclear function in response to SiO2 nanoparticles. AB - Despite of their exponentially growing use, little is known about cell biological effects of nanoparticles. Here, we report uptake of silica (SiO(2)) nanoparticles to the cell nucleus where they induce aberrant clusters of topoisomerase I (topo I) in the nucleoplasm that additionally contain signature proteins of nuclear domains, and protein aggregation such as ubiquitin, proteasomes, cellular glutamine repeat (polyQ) proteins, and huntingtin. Formation of intranuclear protein aggregates (1) inhibits replication, transcription, and cell proliferation; (2) does not significantly alter proteasomal activity or cell viability; and (3) is reversible by Congo red and trehalose. Since SiO(2) nanoparticles trigger a subnuclear pathology resembling the one occurring in expanded polyglutamine neurodegenerative disorders, we suggest that integrity of the functional architecture of the cell nucleus should be used as a read out for cytotoxicity and considered in the development of safe nanotechnology. PMID- 15777788 TI - Inhibition of SNARE-mediated membrane traffic impairs cell migration. AB - Cell migration occurs as a highly-regulated cycle of cell polarization, membrane extension at the leading edge, adhesion, contraction of the cell body, and release from the extracellular matrix at the trailing edge. In this study, we investigated the involvement of SNARE-mediated membrane trafficking in cell migration. Using a dominant-negative form of the enzyme N-ethylmaleimide sensitive factor as a general inhibitor of SNARE-mediated membrane traffic and tetanus toxin as a specific inhibitor of VAMP3/cellubrevin, we conducted transwell migration assays and determined that serum-induced migration of CHO-K1 cells is dependant upon SNARE function. Both VAMP3-mediated and VAMP3-independent traffic were involved in regulating this cell migration. Inhibition of SNARE mediated membrane traffic led to a decrease in the protrusion of lamellipodia at the leading edge of migrating cells. Additionally, the reduction in cell migration resulting from the inhibition of SNARE function was accompanied by perturbation of a Rab11-containing alpha(5)beta(1) integrin compartment and a decrease in cell surface alpha(5)beta(1) without alteration to total cellular integrin levels. Together, these observations suggest that inhibition of SNARE mediated traffic interferes with the intracellular distribution of integrins and with the membrane remodeling that contributes to lamellipodial extension during cell migration. PMID- 15777789 TI - Rnd3/RhoE induces tight junction formation in mammary epithelial tumor cells. AB - Glucocorticoid hormones stimulate adherens and tight junction formation in Con8 mammary epithelial tumor cells through a multistep process in which the membrane organization of structural apical junction proteins and tight junction sealing is controlled by specific signal transduction components. We have previously shown that dexamethasone stimulation of apical junction formation requires down regulation of the small GTPase RhoA. Here we identified Rnd3/RhoE, a GTPase deficient Rho family member and RhoA antagonist, as a key regulator of apical junction dynamics. Exogenously expressed Rnd3/RhoE co-localized with actin at the cell periphery and induced the localization of the adherens junction protein beta catenin and the tight junction protein ZO-1 to sites of cell-cell contact, and led to the formation of highly sealed tight junctions. Treatment with glucocorticoids was not required to achieve complete apical junction remodeling. Consistent with Rnd3/RhoE acting as an antagonist of RhoA, expression of Rnd3/RhoE rescued the disruptive effects of constitutively active RhoA on apical junction organization. Our results demonstrate a new role for the Rho family member Rnd3/RhoE in regulating the assembly of the apical junction complex and tight junction sealing. PMID- 15777790 TI - Cell density-dependent regulation of matrix metalloproteinase and TIMP expression in differently tumorigenic breast cancer cell lines. AB - Matrix metalloproteinases (MMPs) play a key role in cellular invasion and growth. Recent observations on tumor tissue samples suggest that MMP activity is altered in relation to cell density. Therefore, we examined MMP(-1,-2,-3,-8,-9,-10,-11 and -13) and TIMP-1/-2 expression of well-defined cell densities in breast carcinoma cell lines with differing in vivo tumorigenicity/invasiveness (MCF-7 < MDA-MB-468 < MDA-MB-231 < MDA-MB-435). Chemoinvasion assays were performed to link the in vitro data to the in vivo behavior. In accord with previous in vivo data, expression levels of most MMPs decreased significantly with increasing cell density and correlated well with a lower in vitro invasiveness of confluent cells. Since these data suggested that cell density regulates transcription and the promoter regions of most MMPs have AP-1 transcription factor binding consensus sequences, we tested whether functional AP-1 protein was involved in the mechanism of MMP downregulation by cell density. A role for AP-1 was confirmed by over-expression of c-Jun and c-fos in confluent MDA-MB-231 cells, showing with c-Jun increased MMP-2 (5-fold), MMP-3 (1.6-fold), and MMP-9 (160 fold) expression, as well as enhanced invasive potential, while TIMP-1 expression was down-regulated (2-fold) when compared to vector controls. Our data provide clear evidence that cell density regulates major MMPs and TIMPs which are controlled by AP-1 activity so that ultimately a major regulation pathway for the control of the invasive potential of tumor cells is presented. PMID- 15777791 TI - Transient expression of laminin alpha1 chain in regenerating murine liver: restricted localization of laminin chains and nidogen-1. AB - Most interstitia between epithelial and endothelial cells contain basal laminae (BLs), as defined by electron microscopy. However, in liver, the sinusoidal interstitium (called space of Disse) between hepatocytes and sinusoidal endothelial cells (SECs) lacks BLs. Because laminins are major components of BLs throughout the body, whether laminins exist in sinusoids has been a controversial issue. Despite recent advances, the distribution and expression of laminin chains have not been well defined in mammalian liver. Here, using a panel of antibodies, we examined laminins in normal and regenerating mouse livers. Of alpha chains, alpha5 was widely observed in all BLs except for sinusoids, while the other alpha chains were variously expressed in Glisson's sheath and central veins. Laminin gamma1 was also distributed to all BLs except for sinusoids. Although the beta2 chain was observed in all BLs and sinusoids, the expression of beta1 chain was restricted to Glisson's sheath. Detailed analysis of regenerating liver revealed that alpha1 and gamma1 chains appeared in sinusoids and were produced by stellate cells. The staining of alpha1 and gamma1 chains reached its maximum intensity at 6 days after two-thirds partial hepatectomy (PHx). Moreover, in vitro studies showed that alpha1-containing laminin promoted spreading of sinusoidal endothelial cells (SECs) isolated from normal liver, but not other hepatic cells. In addition, SECs isolated from regenerating liver elongated pseudopodia on alpha1-containing laminin more so than did cells from normal liver. The transient expression of laminin alpha1 may promote formation of sinusoids after PHx. PMID- 15777792 TI - The intermediate filament protein vimentin binds specifically to a recombinant integrin alpha2/beta1 cytoplasmic tail complex and co-localizes with native alpha2/beta1 in endothelial cell focal adhesions. AB - Integrin receptors are crucial players in cell adhesion and migration. Identification and characterization of cellular proteins that interact with their short alpha and beta cytoplasmic tails will help to elucidate the molecular mechanisms by which integrins mediate bi-directional signaling across the plasma membrane. Integrin alpha2beta1 is a major collagen receptor but to date, only few proteins have been shown to interact with the alpha2 cytoplasmic tail or with the alpha2beta1 complex. In order to identify novel binding partners of a alpha2beta1cytoplasmic domain complex, we have generated recombinant GST-fusion proteins, incorporating the leucine zipper heterodimerization cassettes of Jun and Fos. To ascertain proper functionality of the recombinant proteins, interaction with natural binding partners was tested. GST-alpha2 and GST-Jun alpha2 bound His-tagged calreticulin while GST-beta1 and GST-Fos beta1 proteins bound talin. In screening assays for novel binding partners, the immobilized GST Jun alpha2/GST-Fos beta1 heterodimeric complex, but not the single subunits, interacted specifically with endothelial cell-derived vimentin. Vimentin, an abundant intermediate filament protein, has previously been shown to co-localize with alphavbeta3-positive focal contacts. Here, we provide evidence that this interaction also occurs with alpha2beta1-enriched focal adhesions and we further show that this association is lost after prolonged adhesion of endothelial cells to collagen. PMID- 15777793 TI - The intracellular domain of teneurin-1 interacts with MBD1 and CAP/ponsin resulting in subcellular codistribution and translocation to the nuclear matrix. AB - Teneurin-1 is a type II transmembrane protein expressed in neurons of the developing and adult central nervous system. To investigate the intracellular signaling of teneurin-1, we searched for proteins interacting with its intracellular domain. One of the proteins identified is the c-Cbl-associated protein CAP/ponsin, an adaptor protein containing SH3 domains. This interaction results on one hand in the recruitment of the soluble intracellular domain of teneurin-1 to the cell membrane enriched in CAP/ponsin. On the other hand, it leads to the translocation of CAP/ponsin to the nucleus, the major site of accumulation of the intracellular domain of teneurin-1. The second interacting protein identified is the methyl-CpG binding protein MBD1. In the nucleus, the intracellular domain of teneurin-1 colocalizes with this transcriptional repressor in foci associated with the nuclear matrix. We propose that these interactions are part of a specific signaling pathway. Evidence for cleavage and nuclear translocation of the intracellular domain has been obtained by the detection of endogenous teneurin-1 immunoreactivity in nuclear speckles in chick embryo fibroblasts. Furthermore, in the nuclear matrix fraction of these cells as well as in cells expressing a hormone-inducible full-length teneurin-1 protein, a teneurin-1 fragment of identical size could be detected as in cells transfected with the intracellular domain alone. PMID- 15777794 TI - TEM8 expression stimulates endothelial cell adhesion and migration by regulating cell-matrix interactions on collagen. AB - The TEM8 gene is selectively expressed in tumor versus normal blood vessels, though its function in endothelial cell biology is not known. Towards the goal of clarifying this function, we tested whether TEM8 overexpression, or blocking TEM8's function with a dominant negative protein, would modulate endothelial cell activities. We found that TEM8-expressing endothelial cells migrated at a rate 3 fold greater than control cells in a monolayer denudation assay. Also, the addition of recombinant TEM8 extracellular domain (TEM8-ED) specifically inhibited both chemokinetic and chemotactic migration on collagen in the denudation and Boyden chamber assays, respectively. The TEM8-ED binds preferentially to collagen, and TEM8 expression enhanced endothelial adhesion to collagen 3-fold; the latter response was antagonized by the TEM8-ED. Consistent with the TEM8-ED acting as a dominant negative inhibitor of endogenously expressed protein were data showing that the TEM8-ED had no effect on the activation of beta1 integrin. TEM8 protein is present in human umbilical vein in situ and is expressed in low passage HUVEC in vitro. TEM8 protein expression in HUVEC was increased 5-fold by the initiation of tube formation, correlating expression of TEM8 with the angiogenic response. Taken together, these results indicate that TEM8 plays a positive role in endothelial cell activities related to angiogenesis. PMID- 15777795 TI - Microarray analysis of immune challenged Drosophila hemocytes. AB - Insect hemocytes play multiple roles in immunity and carry out cellular responses like phagocytosis, encapsulation and melanization as well as producing humoral effector proteins in the first line of defense after injury and invasion of microorganisms. In this work, we used the Drosophila melanogaster hemocyte-like cell line mbn-2 and Affymetrix Drosophila GeneChips to investigate the transcriptome of a single type of immune competent tissue exposed to Gram negative cell wall components (crude LPS) or high dose infection with live Escherichia coli. We found that gene expression profiles of both treatments overlap but show important differences in expression levels of several genes involved in immunity. In addition, cell morphology during infection was monitored and revealed distinct alterations in cell shape and adhesion. Presence of large numbers of bacteria also increased the number of cells taking on crystal cell fate. Taken together, our results indicate that hemocytes sense and respond differently to purified bacterial surface molecules and infection with live and actively growing bacteria both at the level of gene expression and in cell behavior. PMID- 15777796 TI - Annexin V and terminal differentiation of growth plate chondrocytes. AB - Terminal differentiation and mineralization are the final events in endochondral bone formation and allow the replacement of cartilage by bone. Retinoic acid (RA) stimulates these events, including upregulation of expression and activity of alkaline phosphatase (APase), expression of annexins II, V, and VI proteins, which bind to membranes and form Ca(2+) channels, expression of osteocalcin and runx2, another mineralization-related protein and terminal differentiation related transcription factor, and ultimately mineralization. Chelating cytosolic Ca(2+) with BAPTA-AM, interfering with annexin Ca(2+) channel activities using K 201, a specific annexin Ca(2+) channel blocker, or suppression of annexin V expression using siRNA inhibited these events. Overexpression of annexin V in embryonic chicken growth plate chondrocytes resulted in an increase of cytoplasmic Ca(2+) concentration, [Ca(2+)](i) similar to [Ca(2+)](i) increase in RA-treated cultures. Overexpression of annexin V also resulted in upregulation of annexin II, annexin VI, osteocalcin, and runx2 gene expression, expression and activity of APase, and ultimately stimulation of mineralization. K-201 inhibited upregulation of osteocalcin and runx2 gene expression, APase expression and activity, and mineralization in annexin V-overexpressing growth plate chondrocytes. These findings indicate that annexins II, V, and VI alter Ca(2+) homeostasis in growth plate chondrocytes thereby regulating terminal differentiation and mineralization events. Overexpression of annexin V is sufficient to stimulate these terminal differentiation events in growth plate chondrocytes, whereas suppression of annexin V expression inhibits these events. PMID- 15777797 TI - The transcriptional repressor DREAM is involved in thyroid gene expression. AB - Downstream regulatory element antagonistic modulator (DREAM) was originally identified in neuroendocrine cells as a calcium-binding protein that specifically binds to downstream regulatory elements (DRE) on DNA, and represses transcription of its target genes. To explore the possibility that DREAM may regulate the endocrine activity of the thyroid gland, we analyzed its mRNA expression in undifferentiated and differentiated thyroid cells. We demonstrated that DREAM is expressed in the normal thyroid tissue as well as in differentiated thyroid cells in culture while it is absent in FRT poorly differentiated cells. In the present work, we also show that DREAM specifically binds to DRE sites identified in the 5' untranslated region (UTR) of the thyroid-specific transcription factors Pax8 and TTF-2/FoxE1 in a calcium-dependent manner. By gel retardation assays we demonstrated that thapsigargin treatment increases the binding of DREAM to the DRE sequences present in Pax8 and TTF-2/Foxe1 5' UTRs, and this correlates with a significant reduction of the expression of these genes. Interestingly, in poorly differentiated thyroid cells overexpression of exogenous DREAM strongly inhibits Pax8 expression. Moreover, we provide evidence that a mutated form of DREAM unable to bind Ca(2+) interferes with thyroid cell proliferation. Therefore, we propose that in thyroid cells DREAM is a mediator of the calcium-signaling pathway and it is involved in the regulation of thyroid cell function. PMID- 15777798 TI - C/EBP alpha and beta mimic retinoic acid activation of IGFBP-5 in neuroblastoma cells by a mechanism independent from binding to their site. AB - Signal transduction mediated by insulin-like growth factors is implicated in the aggressive behavior of neuroblastoma (NB), a childhood tumor originating from the neural crest. IGFBP-5, a protein that binds IGFs with high affinity, is expressed in many NB cell lines exerting opposite effects, depending on its concentration. We found that IGFBP-5 expression increased during retinoic acid (RA)-mediated differentiation of NB cells. This was due to transcriptional activation as demonstrated by reporter assays carried out in basal and differentiating conditions. We defined the shortest region of the human IGFBP-5 promoter (from nucleotide -83 to +53) which is sensitive to RA. Mutation of a CCAAT enhancer binding protein (C/EBP) element inside this region increased transcription, suggesting a repressive role of this sequence. DNA Affinity Precipitation Assays (DAPA) and chromatin immunoprecipitation demonstrated that the binding of C/EBPalpha and beta to the C/EBP site decreased upon treatment with RA. C/EBPalpha and beta induced an increase in IGFBP-5 transcription in human and murine NB cells similar to that obtained upon RA treatment. Activation by C/EBP alpha and beta did not depend on their binding to the C/EBP site, since they still activated IGFBP-5 promoter carrying a mutation in the C/EBP site. Of interest, we found that both transcription factors were able to interact with the TATA box, but only C/EBPalpha interaction increased during RA-induced differentiation. PMID- 15777799 TI - Hydrogen peroxide inhibits formation of cartilage in chicken micromass cultures and decreases the activity of calcineurin: implication of ERK1/2 and Sox9 pathways. AB - Calcineurin was found as a positive regulator of chondrogenesis in chondrifying chicken micromass cultures (HDCs), as cyclosporine A (CsA) reduced both the amount of cartilage and the expression of mRNAs of aggrecan and the chondrogenic transcription factor Sox9. Cartilage formation was inhibited by H(2)O(2) in a concentration-dependent manner without loss of cellular viability or severe decrease of cell number. Expression of both the mRNA and the unphosphorylated protein Sox9 was decreased, while its phosphorylation was stimulated by either H(2)O(2) or CsA. Oxidative stress decreased the activity of calcineurin but the phosphorylation of the member of MAPK family ERK1/2 was extremely elevated either by 1 mM H(2)O(2) or 2 muM CSA. The ERK inhibitor PD098059 attenuated the depletion of cartilage matrix as well as decreased the expression and phosphorylation of Sox9 in cultures treated with H(2)O(2) or CsA. Our results suggest that the chondrogenesis-inhibiting effect of H(2)O(2) is mediated, at least partly, by inhibition of calcineurin and by activation of ERK1/2. We also propose a regulatory role of calcineurin in the phosphorylation level of either ERK1/2 or Sox9 and a positive role of ERK1/2 in regulating both the expression level and the phosphorylation state of Sox9 in chicken HDCs. PMID- 15777800 TI - PKC 412 sensitizes U1810 non-small cell lung cancer cells to DNA damage. AB - Non-small cell lung carcinoma (NSCLC) is characterized by resistance to drug induced apoptosis, which might explain the survival of lung cancer cells following treatment. Recently we have shown that the broad-range kinase inhibitor staurosporine (STS) reactivates the apoptotic machinery in U1810 NSCLC cells [Joseph et al., Oncogene 21 (2002) 65]. Lately, several STS analogs that are more specific in kinase inhibition have been suggested for tumor treatment. In this study the apoptosis-inducing ability of the STS analogs PKC 412 and Ro 31-8220 used alone or in combination with DNA-damaging agents in U1810 cells was investigated. In these cells Ro 31-8220 neither induced apoptosis when used alone, nor sensitized cells to etoposide treatment. PKC 412 as a single agent induced death of a small number of U1810 cells, whereas it efficiently triggered a dose- and time-dependent apoptosis in U1285 small cell lung carcinoma cells. In both cell types PKC 412 triggered release of mitochondrial proteins followed by caspase activation. However, concomitant activation of a caspase-independent pathway was essential to kill NSCLC cells. Importantly, PKC 412 was able to sensitize etoposide- and radiation-induced death of U1810 cells. The best sensitization was achieved when PKC 412 was administered 24 h after treatments. In U1810 cells, Ro 31-8220 decreased PMA-induced ERK phosphorylation as efficiently as PKC 412, indicating that the failure of Ro 31-8220 to induce apoptosis was not due to weaker inhibition of conventional and novel PKC isoforms. However, Ro 31-8220 increased the basal level of ERK and Akt phosphorylation in both cell lines, whereas Akt phosphorylation was suppressed in the U1810 cells, which might influence apoptosis. These results suggest that PKC 412 could be a useful tool in increasing the efficiency of therapy of NSCLC. PMID- 15777801 TI - Src tyrosine kinase-dependent migratory effects of antithrombin in leukocytes. AB - Tyrosine kinases are known to play a critical role in the regulation of leukocyte function. Antithrombin mediates its effects via syndecan-4 which is known to be linked to the Src tyrosine kinases. In this study, we investigated the role of Src tyrosine kinases in antithrombin-regulated leukocyte migration and Src tyrosine kinase phosphorylation in response to stimulation with antithrombin. Neutrophils and monocytes obtained from forearm venous blood were pre-treated by various Src-family selective tyrosine kinase inhibitors with or without antithrombin followed by washing and assessment of their migratory response toward antithrombin, interleukin-8, or RANTES using Boyden microchemotaxis chambers. Activation status of the two major Src tyrosine kinase phosphorylation sides Tyr416 and Tyr527 was tested using Western blot analysis. Dose-dependent reversal of the antithrombin-mediated effects on neutrophil and monocyte migration was induced by the selective Src kinase inhibitors PP1 and PP2. In Western blot analyses, antithrombin increased Tyr416 and decreased Tyr527 phosphorylation of Src tyrosine kinases in a time- and dose-dependent manner. Moreover, co-incubation with antithrombin lowered the level of RANTES-induced Tyr416 phosphorylation. Therefore, Src tyrosine kinases linked to signaling of antithrombin-binding sites on leukocytes may play an important role in modulating effects on cells function. PMID- 15777802 TI - Responses of keratinocytes to substrate-bound vitronectin: growth factor complexes. AB - Insulin-like growth factor-1 (IGF-I) can associate with the extracellular matrix protein vitronectin (VN) via select IGF-binding proteins, and the resulting complex stimulates responses in a variety of cell types. As VN can also associate with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF), we hypothesized that the multimeric nature of VN could be exploited to deliver multiple growth factors to the cell surface. We report here that VN enhances bFGF but not EGF stimulated [(3)H]-leucine incorporation in the HaCAT keratinocyte cell line, with VN synergistically enhancing cell migration in response to both EGF and bFGF when presented as a VN-bound complex. Furthermore, the addition of EGF and/or bFGF to IGF-I:IGFBP-5:VN complexes significantly enhances both [(3)H] leucine incorporation and migration of HaCAT cells above that induced by IGF:IGFBP-5:VN complexes alone. Indeed, similar responses are observed in primary cultures of human skin keratinocytes, highlighting the potential use of these novel complexes for a wide range of tissue repair applications. PMID- 15777803 TI - Women's attractiveness judgments of self-resembling faces change across the menstrual cycle. AB - Two lines of reasoning predict that women's preferences for people exhibiting cues to kinship will be lower in the follicular phase than in the luteal phase of the menstrual cycle. Women may avoid kinship cues during the follicular phase when they are most fertile due to the costs of inbreeding. Alternatively, women may seek kinship cues during the luteal phase as a byproduct of the benefits of associating with kin during pregnancy, which is also characterized by high progesterone. We find that preferences for facial resemblance, a putative kinship cue, follow this predicted pattern and are positively correlated with estimated progesterone levels based on cycle day. Neither estimated estrogen levels nor conception risk predicted preferences for self-resemblance, and the cyclic shift was stronger for preferences for female faces than male faces. These findings lead to the possibility that this cyclic change in preference for self resemblance may be a byproduct of a hormonal mechanism for increasing affiliative behavior toward kin during pregnancy rather than a mechanism for preventing inbreeding during fertile periods. PMID- 15777804 TI - New evidence that the MHC influences odor perception in humans: a study with 58 Southern Brazilian students. AB - Increasing evidence suggests a correlation between mate choice, odor preference, and genetic similarity at the Major Histocompatibility Complex (MHC) in a variety of animals, including our species. The MHC is a highly polymorphic group of genes that play an important role in the immunological self/nonself recognition. Its products have been reported to take part on the variety of compounds and reactions that together build an individual's body odor. It has been suggested, therefore, that animals use body odor as a guide to identify possible mates as MHC-similar or MHC-dissimilar from their own genotype. Preference for a MHC dissimilar partner enhances MHC heterozygosity of an individual's offspring. The possible adaptive advantages are clear: it is a mechanism of avoiding inbreeding and MHC-heterozygous offspring may have enhanced immunocompetence. The aim of this study was to search, in our species, new evidence on the correlation between specificities at HLA-A and HLA-B and assessments of pleasantness regarding specific body odors. HLA is the name for the human MHC. Four olfactory sessions were performed with 58 young Southern Brazilian students, in order to investigate whether assessments of pleasantness of body odors from individuals correlate to a person's HLA phenotype. Body odors were collected via sweat and urine from all participants. Women smelled and scored all male odor samples and men did the same with all female samples. We found a significant correlation only when female smellers evaluated male sweat odors. PMID- 15777805 TI - Testosterone, testes size, and mating success in birds: a comparative study. AB - Reproductive behaviors of vertebrates are often underpinned by temporal patterns of hormone secretion. We investigated interspecific patterns of circulating testosterone in male birds to test the hypothesis that testosterone plays a crucial role in sexual selection as determined by degree of polygyny and extra pair paternity. We predicted that the evolution of increased levels of polygyny and extra-pair paternity would have resulted in the evolution of increased levels of testosterone to allow males more efficiently to compete for mates. This hypothesis was tested in comparative analyses of 116 species of birds using Generalized Least Squares Models. We assessed the importance of latitudinal distribution, because this can confound the relationship between testosterone and mating success. There were weak positive phylogenetic correlations between measures of testosterone and estimates of mating success at the social level, but this association appeared to be confounded by latitudinal distribution, a significant correlate of testosterone titers. However, we found a significantly positive relationship between peak and residual peak testosterone (which is the peak testosterone level that is controlled for the baseline level) and extra-pair paternity independent of latitude. These results suggest that selection pressures arising from social and sexual mating differently affected testosterone levels with the former being mediated by factors associated with latitudinal distribution. An analysis of residual testes size revealed a positive association between peak and residual testosterone and testes size relative to body size. In a path analysis, we show that relative testis size primarily evolved in association with intense sperm competition and thus high sperm production, and these mechanisms had a secondary impact on blood testosterone levels at a phylogenetic scale. Our results suggest that sperm competition has played an important role in the evolution of reproductive mechanisms in birds. PMID- 15777806 TI - Estradiol and progesterone in paternal and non-paternal hamsters (Phodopus) becoming fathers: conflict with hypothesized roles. AB - Phodopus campbelli has an extensive paternal behavior repertoire whereas the closely-related Phodopus sungorus is not paternally responsive to a displaced pup. For the first time in a naturally paternal mammal, male estradiol and progesterone were determined during two critical phases: (1) the transition from sexually naive male to paired, expectant father that occurs in the absence of stimuli from pups (sexually naive males, paired males on G8, G12, G15, or G17 of the 18-day gestation) and (2) after pup stimuli became available to the males (paired males on days L1, L3, L5, or L12 of pup development). Hormone concentrations in naive males and between G17 and L1 (as stimuli from the birth and the pups became available to males) were also compared. Paternal responsiveness was tested on L3-L5 and confirmed species differences. Hormone concentrations in naive males were similar in the two species and males of both species had estradiol concentrations as high as fertile adult females. However, in direct contrast to predictions, estradiol concentrations were stable in P. campbelli males but increased before the birth, fell across the birth, and increased over pup development in P. sungorus males. Progesterone concentrations in P. campbelli males increased from G17 to L1 whereas a decrease had been predicted. Testosterone dynamics were consistent with previous studies. Either hormonal facilitation of paternal behavior is a hyper-variable trait that has evolved differently in different species, or, more probably, peripheral hormone concentrations are inadequate to explain the role of sex steroid hormones in paternal behavior. PMID- 15777807 TI - Corticosterone suppresses immune activity in territorial Galapagos marine iguanas during reproduction. AB - Individuals that display elaborate sexually selected characters often show reduced immune function. According to the immunocompetence handicap hypothesis, testosterone (T) is responsible for this result as it drives the development and maintenance of sexual characters and causes immunosuppression. But glucocorticoids also have strong influences on immune function and may also be elevated in reproductively active males. Here, we compared immune activity using the phytohemagglutinin (PHA) skin test in three discrete groups of male marine iguanas (Amblyrhynchus cristatus): territorials, satellites, and bachelors. Males of these three reproductive phenotypes had indistinguishable T concentrations during the height of the breeding season, but their corticosterone (cort) concentrations, body condition and hematocrit were significantly different. Territorial males, the animals with the most elaborate sexual ornaments and behaviors, had lower immune responses and body condition but higher cort concentrations and hematocrit than satellites or bachelors. To test directly cort's immunosuppressive role, we elevated cort by either restraining animals or additionally injecting cort and compared their PHA swelling response with the response of free-roaming animals. Such experimental elevation of cort significantly decreased immune activity in both restrained and cort-injected animals. Our data show that cort can induce immunosuppression, but they do not support the immunocompetence handicap hypothesis in its narrow sense because T concentrations were not related to immunosuppression. PMID- 15777808 TI - Prenatal stress differentially affects habituation of corticosterone responses to repeated stress in adult male and female rats. AB - Environmental factors operating early in life have long-lasting and important consequences for the mental and physical health of the adult organism. In particular, prenatal exposure to stress represents one category of adverse early environmental events that are associated with development of depression and schizophrenia in adulthood. In the present studies, we examined whether prenatal stress alters the habituation of hypothalamic-pituitary-adrenal (HPA) activity that occurs with repeated stress exposure in adulthood. We compared corticosterone responses to the first vs. the eighth restraint, with lower responses to the eighth vs. the first considered evidence of habituation. In males, prenatal stress prevented the habituation of corticosterone responses to repeated restraint that was observed in non-prenatally stressed rats. Limited evidence of habituation was seen in either group of females and prenatally stressed females did not exhibit the enhanced corticosterone response during recovery from the eighth restraint that was seen in non-prenatally stressed females. Together, these results suggest a sex-specific interaction between prenatal stress and adult chronic stress on HPA activity. PMID- 15777809 TI - Normally occurring intersexuality and testosterone induced plasticity in the copulatory system of adult leopard geckos. AB - The copulatory neuromuscular system of lizards is highly sexually dimorphic. Adult males possess bilateral penises called hemipenes, which are independently controlled by two muscles, the retractor penis magnus (RPM) and transversus penis (TPN). These structures are not obvious in adult females. However, in adult female leopard geckos (Eublepharis macularius), testosterone induces hemipene growth. We investigated whether these structures develop de novo in adulthood or are histologically present as rudimentary structures in the female leopard gecko. We also investigated the extent of sexual dimorphisms and plasticity in the associated neuromuscular components. To do this, we compared copulatory morphology (sizes of hemipenes, RPM and TPN muscle fibers, and associated motoneurons, as well as motoneuron and RPM fiber number) in adult females treated with testosterone, control females, and control males. All of the geckos possessed hemipenes, RPMs and TPNs, but these structures were indeed vestigial in control females. Testosterone induced striking increases in hemipene and copulatory muscle fiber size in females, but not to levels equivalent to control males. In parallel, males with increased levels of androgenic activity had larger hemipenes, suggesting naturally occurring steroid-induced plasticity. Copulatory motoneurons were not sexually dimorphic in size or number, and these measures did not respond to testosterone. The data demonstrate that the copulatory system of leopard geckos, in which gonadal sex is determined by egg incubation temperature, differs from that of many species (both reptilian and mammalian) with genotypic sex determination. Indeed, the system is remarkable in that adult females have normally occurring intersex characteristics and they exhibit substantial steroid induced morphological plasticity in adulthood. PMID- 15777810 TI - Testosterone implants alter the frequency range of zebra finch songs. AB - To investigate whether changes in testosterone alter the frequency range in which a zebra finch produces song, we assigned male zebra finches to two groups, one of which received testosterone implants and the other empty silastic capsule implants. We then recorded songs up to 52 weeks after the surgery and measured frequency changes in the fundamental frequencies of arbitrarily chosen harmonic stacks in the songs of birds in either group. We found statistically significant decreases after 5 weeks in the songs of the testosterone-treated birds. No changes were found in the fundamental frequencies of the control group. The frequency change remained after the apparent effects of the testosterone implants ended. These data show that high levels of testosterone can lower the frequencies of elements in zebra finch songs. PMID- 15777811 TI - The interaction of testosterone and breeding phase on the reproductive behavior and use of space of male zebra finches. AB - It is well known that androgens play a critical role in mediating the reproductive behavior of males. However, many laboratory experiments that examined the effects of testosterone in male songbirds typically limited their investigations to the early phase of breeding. We sought to determine the influence of testosterone on social behavior, pair bonding, nesting, and use of space in captive zebra finch (Taeniopygia guttata) males as a function of breeding stage (pre-laying, incubation, and nestling phases). Fourteen males were released into an aviary with 14 females and allowed to breed for 7 weeks. Half of the males were given testosterone implants and half were given control implants. During the pre-laying phase, testosterone-implanted males spent significantly more time in nesting activities than control birds and more time elapsed from starting to build a nest to when their mates initiated egg-laying. During the incubation phase, testosterone-implanted subjects spent significantly more time in female-directed and undirected singing. Use of space varied between hormone conditions depending upon breeding phase: there was no difference during pre laying, but during the incubation and nestling phases, testosterone-implanted subjects used significantly more space. This significant increase in "home range" during the latter phase of the breeding cycle coincides with results from field studies on other species. These results underscore the importance of considering breeding phase in assessing the behavioral sensitivity to hormones. PMID- 15777812 TI - Modulation of prolactin but not corticosterone responses to stress in relation to parental effort in a long-lived bird. AB - We tested the hypothesis that parental effort modulates the magnitude of corticosterone and prolactin responses to stress in a long-lived bird, the Black legged kittiwake (Rissa tridactyla). To do so, we compared corticosterone and prolactin responses to capture/restraint stress between chick-rearing birds and failed breeders (no parental effort). We predicted that (1) the increase in plasma corticosterone levels in response to stress should be lower in chick rearing birds, (2) the decrease in plasma prolactin levels in response to stress should be lower in chick-rearing birds, and (3) as both sexes care for the chick, there should be no sex difference in the hormonal response to stress. Baseline plasma corticosterone and prolactin levels were higher in chick-rearing birds and were not influenced by body condition. Failed breeders were in better condition than chick-rearing individuals. Corticosterone response to stress was unaffected by parental effort as both chick-rearing and failed birds exhibited a robust corticosterone increase. Prolactin response to stress was however clearly influenced by parental effort: chick-rearing birds showed a modest 9% prolactin decrease whereas in failed birds prolactin concentrations fell by 41%. Body condition did not influence hormonal responses to stress. When facing stressful condition, breeding kittiwakes attenuate their prolactin response to stress while enhancing their secretion of corticosterone. Increasing corticosterone secretion triggers foraging efforts and diminishes nest attendance whereas an attenuation of prolactin response to stress maintains parental behavior. We suggest that this hormonal mechanism facilitates a flexible time-budget that has been interpreted as a buffer against environmental variability. PMID- 15777813 TI - Coordinated and dissociated effects of testosterone on singing behavior and song control nuclei in canaries (Serinus canaria). AB - Temperate zone songbirds that breed seasonally exhibit pronounced differences in reproductive behaviors including song inside and outside the breeding season. Springlike long daylengths are associated with increases in plasma testosterone (T) concentrations, as well as with increases in singing and in the volume of several brain nuclei known to control this behavior. The mechanisms whereby T can induce changes in behavior and brain, and whether or not these effects are differentially regulated, have recently begun to be examined, as has the question of the relative contributions of T and its androgenic and estrogenic metabolites to the regulation of this seasonal behavioral and neural plasticity. In this experiment, we examined the effects of T, 5alpha-dihydrotestosterone, or 17beta estradiol treatment on castrated male canaries housed on short days and compared neural and behavioral effects in these males to similarly-housed males given only blank implants. We observed that only T treatment was effective in eliciting significant increases in singing behavior after 11 days of hormone exposure. In addition, T alone was effective in increasing the volume of a key song production nucleus, HVC. However, at this time, none of the steroids had any effects on the volumes of two other song control nuclei, Area X of the medial striatum and the robust nucleus of the arcopallium (RA), that are efferent targets of HVC, known to be regulated by androgen in canaries and also to play a role in the control of adult song. T can thus enhance singing well before concomitant androgen-induced changes in the song control system are complete. PMID- 15777814 TI - Effects of progesterone blockade over cocaine-induced genital reflexes of paradoxical sleep-deprived male rats. AB - Paradoxical sleep deprivation (PSD) enhances cocaine-induced genital reflexes (penile erection [PE] and ejaculation [EJ]) in male rats and induces a significant increase in progesterone concentration. As progesterone treatment facilitates PE in PSD castrated rats, we may speculate that progesterone appears to be a relevant hormonal factor eliciting genital reflexes in PSD males. In order to expand the latter finding, different doses of antiprogestin mifepristone (vehicle, 2.5, 5, 10, and 20 mg/kg, s.c.) were administered to PSD rats at the end of a 4-day period of PSD 1 h prior to cocaine administration (7 mg/kg, i.p.) and placed in observation cages for the evaluation of genital reflexes. Pretreatment with vehicle induced PE in all rats and this effect was significantly reduced by mifepristone at 5 to 20 mg/kg doses that lowered the proportion to 40% of the rats. The frequency of PE was also significantly reduced for all doses used. There were no significant differences between vehicle and mifepristone in EJ behavior. As for hormone concentrations, mifepristone reduced progesterone concentrations at the 5-20 mg/kg doses compared to vehicle group. At 20 mg/kg, it also elevated testosterone concentrations. In addition, mifepristone administration induced a significant decrease in the duration of PS episodes at all doses. These data suggest that progesterone exerts an essential role in erectile response induced by cocaine in PSD male rats. PMID- 15777815 TI - Linear social dominance hierarchy and corticosterone responses in male mallards and pintails. AB - In winter, dabbling ducks gather in large flocks and males compete not only for food resources but also for mates. Setting up a social hierarchy is one way to reduce the costs of conflicts, but the position in the hierarchy has implications for individuals, for instance their susceptibility to conflict and interference, hence to social stress. We investigated relationships between linear social dominance and corticosterone levels, baseline levels and induced-stress response, in captive male mallards Anas platyrhynchos and pintails Anas acuta during the winter period. We hypothesised that corticosterone responses would reflect the costs associated with social stress. From previous work on dominance and corticosterone in wintering birds, we expected that, where the social hierarchy is linear, there would be (1) no relationship between social ranks and baseline corticosterone levels and (2) a significant positive relationship between dominance ranks and responses to acute stress. Our results demonstrated the existence of a linear hierarchy in both species (h' = 0.95 for mallards and h' = 0.97 for pintails), and we found that pintails had on average more corticosterone than mallards. The relationship between dominance and corticosterone responses followed the predictions, with no differences for baseline levels and an attenuated response to induced-stress for subordinates. We discuss these results in the perspective of the cost-benefits of dominance and wintering strategies of waterbirds. PMID- 15777816 TI - Ethology and the origins of behavioral endocrinology. AB - The neurosciences embrace many disciplines, some long established, others of more recent origin. Behavioral endocrinology has only recently been fully acknowledged as a branch of neuroscience, distinctive for the determination of some of its exponents to remain integrative in the face of the many pressures towards reductionism that so dominate modern biology. One of its most characteristic features is a commitment to research at the whole-animal level on the physiological basis of complex behaviors, with a particular but by no means exclusive focus on reproductive behavior in all its aspects. The search for rigorously defined principles of behavioral organization that apply across species and the hormonal and neural mechanisms that sustain them underlies much of the research. Their aims are much like those put forth in the classical ethology of Lorenz and Tinbergen, one of the roots from which behavioral endocrinology has sprung. But there are others that can be traced back a century or more. Antecedents can be found in the work of such pioneers as Jakob von Uexkull, Jacques Loeb, Herbert Spencer Jennings, and particularly Charles Otis Whitman who launched a tradition that culminated in the classical contributions of Robert Hinde and Daniel Lehrman. William C. Young was another pioneer. His studies revolutionized thinking about the physiological mechanisms by which hormones influence behavior. An earlier potent influence was Karl Lashley who helped to shape the career of Frank Ambrose Beach who, more than anyone, has played a leading role in launching this new field. PMID- 15777818 TI - Perinatal risk management: obstetric methods to prevent birth asphyxia. AB - Because obstetric care frequently is associated with the potential for liability, the purpose of this article is to familiarize the reader with perinatal risk management using the concept of foreseeability of harm and its potential application to obstetric care. At the same time, this article introduces the concept of notice, and explains the critical conduct intervals that are used to gauge how well the health care teams handle obstetric emergencies. The focus then shifts to incorporate these concepts into several maternal-child health quality management programs. It is hoped that this article will result in an improvement of perinatal outcome for pregnant women and their unborn children. PMID- 15777819 TI - Management of birth injuries. AB - Birth injuries are a significant cause of neonatal morbidity and mortality. Although often associated with traumatic delivery, birth injuries often occur in normal spontaneous deliveries in the absence of any risk factors. This article discusses the diagnosis and management of the most common birth injuries that are encountered by health care providers who care for newborns. PMID- 15777820 TI - Birth injuries of the brachial plexus. AB - Birth injuries of the brachial plexus are fairly common, but the majority of affected newborns make quick recoveries without any specific intervention. A minority suffer more severe injuries that lead to varying degrees of life-long disability. Happily, modern microsurgical techniques permit reconstruction of certain plexus injuries and, in carefully selected patients, can restore voluntary activity to target muscle groups. To what degree reanimation of paralyzed muscles improves function and quality of life for these children is a more important matter that has not yet been addressed at the level of modern standards of evidence. Brachial plexus reconstruction is only a first step in the multidisciplinary process needed to optimize long-term functional outcomes for severely affected infants. PMID- 15777821 TI - Birth asphyxia and cerebral palsy. AB - There has been a societal presumption that most, if not all, cases of hypoxic ischemic encephalopathy-induced cerebral palsy occur during the 3 hours that are related to the events of labor and delivery; society has tended to overlook the remaining 7000 hours of the pregnancy. As a result of this societal perspective, often times the obstetrician has been targeted unfairly as the person who is responsible for a given child's neurologic injuries. Rather, the entire pregnancy, labor, delivery, and well beyond birth require examination to understand fully the pathophysiologic mechanisms that are responsible for an infant's brain injuries, and their long-term impact on the child. PMID- 15777822 TI - Pitfalls in neonatal resuscitation. AB - Ten percent of all newborns require resuscitation at birth. The Neonatal Resuscitation Program establishes the authoritative technique of newborn resuscitation. Errors continue to occur that are related to the use of unskilled resuscitators; intubation; inadequate suctioning of meconium; and the postresuscitation problems of hypoglycemia, hypocarbia, and hypotension. Specific recommendations are offered to avoid these pitfalls of neonatal resuscitation. PMID- 15777823 TI - Patient safety in the neonatal intensive care unit. AB - Patient safety has become a national priority. This article reviews issues of patient safety in neonatal intensive care. Safety goals for 2005 set by the Joint Commission on Accreditation of Healthcare Organizations are discussed from the perspective of relevance to neonatal intensive care. PMID- 15777824 TI - Prevention of medication errors. AB - Medication error is the most frequent source of medical error that is associated with adverse events, and, in many cases, is preventable. Medication errors can occur at any step in the medication process. Medication error prevention and reduction begins with a systematic approach to their detection. An important approach to mitigating errors involves the reduction of variation in task performance using tested techniques and technologies from other industries. The most important component of error prevention and reduction is the proactive promotion of a safety culture by organizational leadership, with sustained education and support for users. PMID- 15777825 TI - Risk management of severe neonatal hyperbilirubinemia to prevent kernicterus. AB - Our approach for risk management of severe neonatal hyperbilirubinemia to prevent kernicterus--one of the most easily preventable causes of neonatal brain damage- includes management of certain high-risk clinical situations, identification of systems failure, and suggestions for implementation strategies to enhance patient safety. PMID- 15777826 TI - Intravascular catheter complications in the neonatal intensive care unit. AB - Central and peripheral intravascular catheters serve many important functions for sick neonates; however, the use of these catheters also carries significant risk of permanent disability or even death. This article examines the risks of catheter-related thromboembolic events, vasospasm, and infection. The article also looks at risks that are specific to umbilical artery catheters, umbilical vein catheters, peripherally-inserted central catheters, peripheral artery catheters, and peripheral intravenous catheters. Specific recommendations are made to reduce the possibility of an intravascular catheter-related complication. PMID- 15777827 TI - Talking about the unthinkable: perinatal/neonatal communication issues and procedures. AB - Clinician-parent conversations in perinatology and neonatology are demanding. They take place over a period of time and ask much of the clinician. Many parents insist that they want thorough explanations so they can understand the medical information and its impact on them. These cases involve copious amounts of medical and resource information and parents cannot assimilate it all at one time; thus, these conversations are extended dialogs. Conversational skills that seem essential for clinicians in this work include giving bad news, sharing information, and expressing empathy. Underlying the conversational skills are those habits that form the complete physician: curiosity that leads one to learn the parents' values; the skill to help parents apply these values as they make difficult decisions; patience to listen to and stick with these suffering people who are full of strong emotions; and finally, the courage and endurance to confront these difficult situations day after day. PMID- 15777828 TI - A defense attorney's perspective on medical negligence litigation. AB - Being named as a defendant in a lawsuit is an unnerving development. Proceeding through the discovery stage of a case can be a daunting prospect. Sitting through a trial, in which one's professional conduct is judged by six or twelve laypersons with little or no medical knowledge, often is gut wrenching. In this day and age, many physicians, including extremely well qualified specialists, often must deal with these realities. To cope best with these events, this article is an attempt to summarize certain legal principles that are relevant to these cases and to emphasize the practical realities that are attendant to medical negligence litigation. PMID- 15777829 TI - A plaintiff attorney's candid view of medical malpractice. AB - The human dynamics of medical malpractice litigation is more nuanced than many doctors suspect. The purpose of this article is to provide some candid insights into the process. The article presents three subjects that doctors are not likely to learn on their own, even if they read all of the law books or had the misfortune of running the gauntlet of their own medical malpractice lawsuit. Those subjects are: (1) the top five reasons why doctors get sued, (2) avoiding less than optimal legal representation, and (3) the importance of nonmedical issues in litigation. PMID- 15777830 TI - Malpractice crisis and reform. AB - Medical liability coverage has become increasingly expensive and scarce since 2001, especially for obstetrics and other high-risk specialties. Physicians, whose own fees are constrained, see this as a crisis needing tort reform to cap payouts. Plaintiffs' lawyers fight caps; they blame insurance trends and call for stronger regulation of insurers and physicians. Doctors' arguments are somewhat better than lawyers' about insurance developments, but the huge tort reform battle wholly ignores the biggest problems--the medical system has too many preventable medical injuries, and the legal system fails to prevent them. It is time to try out better approaches, three of which are discussed in this paper. PMID- 15777831 TI - Malpractice reforms: are they fair? AB - In the debate over medical malpractice reform, the dimension of fairness often is invoked but is poorly understood. This article describes the factors that promote fairness in public policy and then examines whether proposed or enacted tort reforms would be fair. It finds that many of the reforms that are being promoted by perinatologists would be unfair; however, some reforms, including one broad ranging proposal that was put forward by the Institute of Medicine, could satisfy fairness criteria depending on how they were implemented. PMID- 15777832 TI - Evidence-based expert testimony. AB - Expert witnesses undoubtedly know a great deal about topics on which ordinary people lack information. But experts, no less than other people, are subject to predictable memory biases. No plaintiff lawyer can enter the courtroom alone, without a doctor by his or her side to serve as an expert witness. The behavior of the plaintiff's experts (and the behavior of the experts who walk alongside the defense lawyers as well) is the subject of this article. What should guide expert testimony? How should those guidelines be implemented? How has our profession dealt with these issues? PMID- 15777833 TI - Perinatal litigation and related nursing issues. AB - Obstetric and neonatal nurses are expected to provide an abundance of guidance, support, monitoring, and education to women and their babies during and after delivery. Nurses should adhere to standards of professional nursing practice. This will ensure that optimal and safe care is provided for the mother and fetus or neonate. Perinatal nurses are vulnerable to litigation should complications occur. Perinatal nurses are responsible for providing routine assessments as well as initiating and performing emergency interventions. This includes recognition of the symptoms of complications in the mother and the neonate, resuscitation, and activation of the emergency system. Occasionally, nurses are obliged to question the practice of other health care providers. Although perinatal nurses continue to be at risk for malpractice vulnerability, risk reduction techniques are available to them. This article provides the nurse with knowledge of legal proceedings and strategies to reduce liability when caring for pregnant women and newborns. PMID- 15777834 TI - ERK2 is required for FGF1-induced JNK1 phosphorylation in Xenopus oocyte expressing FGF receptor 1. AB - A possible connection between the ERK2 and JNK1 MAP kinases transduction cascades was investigated in Xenopus oocytes expressing FGFR1 stimulated by FGF1. Injection of various inhibitors for the Shc/Grb2/Ras/Mos/MEK/ERK2 cascade blocked FGF1-induced germinal vesicle breakdown (GVBD), as well as ERK2 and JNK1 phosphorylation. JNK1 was found to be activated downstream of ERK2, since injection of an active ERK2 triggered JNK1 phosphorylation and inhibition of ERK2 either by a MEK inhibitor or the MKP3 phosphatase blocked JNK1 phosphorylation. These results demonstrated that in FGFR1 signalling JNK1 phosphorylation depends on ERK2. PMID- 15777835 TI - Protein targeting to the chloroplasts of photosynthetic eukaryotes: getting there is half the fun. AB - The plastids of many algae are surrounded by three or four membranes, thought to be a consequence of their evolutionary origin through secondary endosymbiosis between photosynthetic and non-photosynthetic eukaryotes. Each membrane constitutes a barrier to the passage of proteins, so protein targeting in these complex plastids has an extra level of difficulty when compared to higher plants. In the latter, protein translocation across the two membranes uses multi-protein complexes that together import proteins possessing an N-terminal leader sequence rich in serine and threonine (S/T). In contrast, while targeting to most complex plastids also involves an S/T-rich region, this region is preceded by an N terminal hydrophobic signal peptide. This arrangement of peptide sequences suggests that proteins directed to complex plastids pass through the ER, as do other proteins with hydrophobic signal peptides. However, this simplistic view is not always easy to reconcile with what is known about the different secondary plastids. In the first group, with plastids bounded by three membranes, plastid directed proteins do indeed arrive in Golgi-derived vesicles, but a second hydrophobic region follows the S/T-rich region in all leaders. In the second group, where four membranes completely surround the plastids, it is still not known how the proteins arrive at the plastids, and in addition, one member of this group uses a targeting signal rich in asparagine and lysine in place of the S/T-rich region. In the third group, the fourth bounding membrane is contiguous with the ER, but it is not clear what distinguishes plastid membranes from others in the endomembrane system. Knowing what to expect is important, as genomic sequencing programs may soon be turning up some of the missing pieces in these translocation puzzles. PMID- 15777836 TI - Enhanced LPS-induced TNF alpha production in heat-shocked human promonocytic cells: regulation at the translational/post-translational level. AB - Heat shock proteins (hsps) play an important role in maintaining cellular homeostasis and protecting cells from various insults. Recent evidence also implicates hsps in the regulation of the immune response, particularly the inflammatory process. In the present study, we showed that human promonocytic cells (THP-1) produced elevated levels of tumor necrosis factor alpha (TNF alpha) after incubation with bacterial lipopolysaccharide (LPS) when cells were pre stressed by a mild heat shock (HS) of 42 degrees C (1.5 h) followed by recovery at 37 degrees C (3 h) in comparison with non-stressed cells also stimulated with LPS. This enhanced TNF alpha production was not due to changes in nuclear factor kappaB (NF-kappa B) activation, TNF alpha transcription rates, or mRNA stability. Thus, an effect at the translational or posttranslational level is likely responsible. Elevated production of TNF alpha was not observed when cells were stimulated with LPS immediately after stress or when HS temperature was increased to 43 degrees C. This negative effect of HS is likely due to a harmful effect of temperature. Moreover, enhanced LPS-induced TNF alpha production was not observed after differentiation of promonocytes into macrophage-like cells. Thus, our results show that the stress temperature, recovery period, and differentiation stage of the cell modulate the effect of HS on the inflammatory process. PMID- 15777837 TI - The distinct erythropoietin functions that promote cell survival and proliferation are affected by aluminum exposure through mechanisms involving erythropoietin receptor. AB - Erythropoietin (Epo) promotes the development of erythroid progenitors by triggering intracellular signals through the binding to its specific receptor (EpoR). Previous results related to the action of aluminum (Al) on erythropoiesis let us suggest that the metal affects Epo interaction with its target cells. In order to investigate this effect on cell activation by the Epo-EpoR complex, two human cell lines with different dependence on Epo were subjected to Al exposure. In the Epo-independent K562 cells, Al inhibited Epo antiapoptotic action and triggered a simultaneous decrease in protein and mRNA EpoR levels. On the other hand, proliferation of the strongly Epo-dependent UT-7 cells was enhanced by long term Al treatment, in agreement with the upregulation of EpoR expression during Epo starvation. Results provide some clues to the way by which Epo supports cell survival and growth, and demonstrate that not all the intracellular factors needed to guarantee the different signaling pathways of Epo-cell activation are available or activated in cells expressing EpoR. This study then suggests that at least one of the mechanisms by which Al interfere with erythropoiesis might involve EpoR modulation. PMID- 15777838 TI - Contact of Chlamydophila pneumoniae with type II cell triggers activation of calcium-mediated NF-kappa B pathway. AB - Nuclear factor-kappa B (NF-kappa B) plays an important role in inflammation, proliferation and regulation of apoptosis. The purpose of the present study on type II cells was to investigate whether Chlamydophila pneumoniae contact induces (I) a Ca2+ release, that (II) disrupts F-actin/beta-tubulin cytoskeletal association with NF-kappa B/I kappa B alpha, leading to (III) a subsequent NF kappa B activation. Incubation of rat type II pneumocytes with C. pneumoniae caused an intracellular calcium release within seconds. Confocal laser scanning microscopy (CLSM) revealed that bacterial contact with cell surface leads to a disappearance of the microvilli and disturbs the co-localization between F-actin and NF-kappa B (p65). Using semi-quantitative CLSM, we show that at 10-30 min I kappa B alpha was decreased and p65 or p50 was simultaneously translocated from cytoplasm to the nucleus, resulting in a 19-fold and 17-fold increase versus control cells. During this time no bacteria were internalized into type II cells. The pre-treatment of cells with BAPTA-AM inhibited C. pneumoniae-mediated calcium release. BAPTA-AM or SN50 prevented the C. pneumoniae-induced changes in F-actin cytoskeleton and inhibited NF-kappa B activation. Paclitaxel reduced C. pneumoniae-mediated changes of beta-tubulin cytoskeleton and activation of NF kappa B. These results suggest that calcium-mediated cytoskeleton reorganization is involved in C. pneumoniae-induced NF-kappa B activation in type II cells. PMID- 15777839 TI - Modulation of hepatocyte growth factor induction in human skin fibroblasts by retinoic acid. AB - Topical treatment of skin with all-trans-retinoic acid (ATRA), the major biologically active form of vitamin A, results in hyperproliferation of basal keratinocytes, leading to an accelerated turnover of epidermis cells and thickening of the epidermis, probably via induction of production of paracrine growth factors for keratinocytes in epidermal suprabasal keratinocytes and/or dermal fibroblasts. Since hepatocyte growth factor (HGF) is a factor mitogenic to epidermal keratinocytes secreted from dermal fibroblasts, the effect of ATRA on basal and induced HGF production in human dermal fibroblasts in culture was examined. ATRA alone did not induce HGF production, but it significantly enhanced HGF production induced by the cAMP-elevating agent cholera toxin or the membrane permeable cAMP analog 8-bromo-cAMP. Cholera toxin-induced activation of cAMP responsive element (CRE)-binding protein (CREB) was enhanced by pretreating cells with ATRA for 24 h. In contrast, HGF production induced by epidermal growth factor (EGF) or phorbol 12-myristate 13-acetate (PMA) was potently inhibited by ATRA. These modulatory effects of ATRA were different from the effects of transforming growth factor-beta1 (TGF-beta) and dexamethasone, both of which inhibited HGF production induced by all of the four inducers. Up-regulation of HGF gene expression by cholera toxin and EGF was also enhanced and inhibited, respectively, by ATRA. Both 9-cis-retinoic acid (9-cis-RA) and 13-cis-retinoic acid (13-cis-RA), which are stereo-isomers of ATRA, showed a modulatory effect on HGF induction similar to that of ATRA. These results suggest that ATRA augments the induction of HGF production caused by increased intracellular cAMP. PMID- 15777840 TI - The metabolism of hyaluronan in cultured rabbit growth plate chondrocytes during differentiation. AB - Hyaluronan (HA) is one of the major extracellular matrix components in cartilage. In addition to the biomechanical functions, HA has various important roles in the differentiation of chondrocytes. The purpose of this study was to clarify the nature of HA synthesis during chondrocyte differentiation. Growth plate chondrocytes were isolated from rabbit ribs and cultured in chondrocyte differentiation medium. The amount of HA and HA synthase (HAS) mRNA levels were analyzed for each stage of chondrocyte differentiation by means of high performance liquid chromatography (HPLC) and real-time PCR, respectively. The distribution of HA in cultured chondrocytes was observed by histochemical staining. The amount of HA, ranging widely in size, was increased substantially during the hypertrophic stage. The expression levels of HAS2 and HAS3 mRNAs were low during the matrix-forming stage. HAS2 mRNA level was substantially enhanced at the pre-hypertrophic stage, whereas HAS3 mRNA level exhibited a slight increase. HAS1 mRNA was not detected. The intensity of HA staining was high around the hypertrophic chondrocytes. These results suggest that HA metabolism in chondrocyte differentiation is regulated by the selective expression of HASs, and HAS2 and the related large size-HA may have a certain association with the hypertrophic changes of chondrocytes. PMID- 15777841 TI - Mechanism of DNA binding and localized strand separation by Pur alpha and comparison with Pur family member, Pur beta. AB - Pur alpha is a single-stranded (ss) DNA- and RNA-binding protein with three conserved signature repeats that have a specific affinity for guanosine-rich motifs. Pur alpha unwinds a double-stranded oligonucleotide containing purine rich repeats by maintaining contact with the purine-rich strand and displacing the pyrimidine-rich strand. Mutational analysis indicates that arginine and aromatic residues in the repeat region of Pur alpha are essential for both ss- and duplex DNA binding. Pur alpha binds either linearized or supercoiled plasmid DNA, generating a series of regularly spaced bands in agarose gels. This series is likely due to localized unwinding by quanta of Pur alpha since removal of Pur alpha in the gel eliminates the series and since Pur alpha binding increases the sensitivity of plasmids to reaction with potassium permanganate, a reaction specific for unwound regions. Pur alpha binding to linear duplex DNA creates binding sites for the phage T4 gp32 protein, an ss-DNA binding protein that does not itself bind linearized DNA. In contrast, Pur beta lacking the Pur alpha C terminal region binds supercoiled DNA but not linearized DNA. Similarly, a C terminal deletion of Pur alpha can bind supercoiled pMYC7 plasmid, but cannot bind the same linear duplex DNA segment. Therefore, access to linear DNA initially requires C-terminal sequences of Pur alpha. PMID- 15777842 TI - The pathway for IRP2 degradation involving 2-oxoglutarate-dependent oxygenase(s) does not require the E3 ubiquitin ligase activity of pVHL. AB - Iron regulatory protein 2 (IRP2), a posttranscriptional regulator of iron metabolism, is subjected to iron-dependent degradation by the proteasome. Recent experiments proposed a mechanism involving 2-oxoglutarate-dependent oxygenases. Enzymes of this class, such as prolyl-4-hydroxylases, mediate the oxygen and iron dependent degradation of the hypoxia inducible factor HIF-1alpha, which requires the E3 ubiquitin ligase activity of pVHL. Considering that the pathways for IRP2 and HIF-1alpha degradation share remarkable similarities, we investigated whether pVHL may also be involved in the degradation of IRP2. We show here that IRP2 can interact with pVHL in co-transfection/co-immunoprecipitation assays. Furthermore, pVHL is able to promote the ubiquitination and the decay of transfected IRP2. However, the iron-dependent degradation of endogenous IRP2 is not impaired in VHL deficient cell lines, suggesting that pVHL is not a necessary component of this pathway. PMID- 15777843 TI - Expression of catalase and glutathione peroxidase in renal insufficiency. AB - Chronic renal failure (CRF) is associated with oxidative stress, the precise mechanism of which is yet to be elucidated. The present study was undertaken to investigate in renal insufficiency the expression of catalase and glutathione peroxidase, which play a critical role in antioxidant defense system by catalyzing detoxification of hydrogen peroxide (H2O2) and organic hydroperoxides. Rats were randomly assigned to the CRF (5/6 nephrectomized) and sham-operated control groups and observed for 6 weeks. Renal and thoracic aortic catalase and glutathione peroxidase protein abundance was measured by Western blotting. The enzyme activities in the renal and aortic extracts, hepatic glutathione levels, blood pressure and urinary nitric oxide metabolites (NO(x)) excretion were also measured. Blood pressure and urinary nitric oxide metabolite (NO(x)) excretion were also measured. The CRF group showed a significant down-regulation of both immunodetectable catalase and glutathione peroxidase proteins in the remnant kidney. Catalase activity was also significantly decreased in the remnant kidney whereas glutathione peroxidase activity was not significantly affected. Furthermore, the protein abundance of catalase was unchanged whereas the enzyme activity was significantly decreased in the thoracic aorta of CRF animals compared to the sham-operated controls. By contrast, both the protein abundance and the enzyme activity of glutathione peroxidase were not significantly affected in the aorta of CRF animals compared to the sham-operated controls. This was coupled with marked arterial hypertension, significant reduction of hepatic glutathione levels and urinary NO(x) excretion pointing to increased inactivation and sequestration of NO by superoxide. These events point to the role of impaired antioxidant defense system in the pathogenesis of oxidative stress in CRF. PMID- 15777844 TI - Stable transfection of Acanthamoeba castellanii. AB - A simple method for stable transfection of Acanthamoeba castellanii using plasmids which confer resistance to neomycin G418 is described. Expression of neomycin phosphotransferase is driven by the Acanthamoeba TBP gene promoter, and can be monitored by cell growth in the presence of neomycin G418 or by Western blot analysis. Transfected cells can be passaged in the same manner as control cells and can be induced to differentiate into cysts, in which form they maintain resistance to neomycin G418 for at least several weeks, although expression of neomycin phosphotransferase is repressed during encystment. Expression of EGFP or an HA-tagged EGFP-TBP fusion can be driven from the same plasmid, using an additional copy of the Acanthamoeba TBP gene promoter or a deletion mutant. The TBP-EGFP fusion is localized to the nucleus, except in a small proportion of presumptive pre-mitotic cells. EGFP expression can also be driven by the cyst specific CSP21 gene promoter, which is completely repressed in growing cells but strongly induced in differentiating cells. Transfected cells maintain their phenotype for several weeks, even in the absence of neomycin G418, suggesting that transfected genes are stably integrated within the genome. These results demonstrate the utility of the neomycin resistance based plasmids for stable transfection of Acanthamoeba, and may assist a number of investigations. PMID- 15777845 TI - Blockade of murine erythroleukemia cell differentiation by hypomethylating agents causes accumulation of discrete small poly(A)- RNAs hybridized to 3'-end flanking sequences of beta(major) globin gene. AB - Induction of murine erythroleukemia (MEL) cell differentiation is accompanied by transcriptional activation of globin genes and biosynthesis of hemoglobin. In this study, we observed cytoplasmic accumulation of relatively small RNAs of different size (150-600 nt) hybridized to alpha1 and beta(major) globin DNA probes in MEL cells blocked to differentiate by hypomethylating agents (neplanocin A, 3-deazaneplanocin A and cycloleucine). These RNAs lack poly(A) tail and appear to be quite stable. Search within the 3'-end flanking sequences of beta(major) globin gene revealed the presence of a B1 repeat element, several ATG initiation codons, a GATA-1 consensus sequence and sequences recognized by AP 1/NF-E2 and erythroid Kruppel-like factor (EKLF) transcription factors. These data taken together indicate that exposure of MEL cells to hypomethylating agents promotes accumulation of relatively small discrete RNA transcripts lacking poly(A) tail regardless of the presence or absence of inducer dimethylsulfoxide (DMSO). However, the relative steady-state level of small RNAs was comparatively higher in cells co-exposed to inducer and each one of the hypomethylating agents. Although the orientation of these RNAs has not been established as yet, the possibility these small poly(A)- RNAs which are induced by hypomethylating agents may be involved in the blockade of MEL cell differentiation program is discussed. PMID- 15777846 TI - Hsp90 inhibitor geldanamycin increases hsp70 mRNA stabilisation but fails to activate HSF1 in cells exposed to hydrostatic pressure. AB - High hydrostatic pressure (HP) increases Hsp70 protein and mRNA levels by increasing the mRNA half-life without activation of HSF1 transcription factor. We investigated whether this change in gene expression requires Hsp90, previously shown to regulate hsp70 genes via HSF1. In HeLa cells, both HP and Hsp90 inhibitor geldanamycin (GA) up-regulated Hsp70 expression through mRNA stabilisation. GA, unlike HP, increased HSF1 activation. However, when exposures were used together a marked Hsp70 response was observed with mRNA stabilisation without coincidence of HSF1 activation. Our data suggests that Hsp90 is involved in hsp70 mRNA stabilisation and the HSF1 activation can be suppressed by high HP. PMID- 15777847 TI - 50-Hz extremely low frequency electromagnetic fields enhance cell proliferation and DNA damage: possible involvement of a redox mechanism. AB - HL-60 leukemia cells, Rat-1 fibroblasts and WI-38 diploid fibroblasts were exposed for 24-72 h to 0.5-1.0-mT 50-Hz extremely low frequency electromagnetic field (ELF-EMF). This treatment induced a dose-dependent increase in the proliferation rate of all cell types, namely about 30% increase of cell proliferation after 72-h exposure to 1.0 mT. This was accompanied by increased percentage of cells in the S-phase after 12- and 48-h exposure. The ability of ELF-EMF to induce DNA damage was also investigated by measuring DNA strand breaks. A dose-dependent increase in DNA damage was observed in all cell lines, with two peaks occurring at 24 and 72 h. A similar pattern of DNA damage was observed by measuring formation of 8-OHdG adducts. The effects of ELF-EMF on cell proliferation and DNA damage were prevented by pretreatment of cells with an antioxidant like alpha-tocopherol, suggesting that redox reactions were involved. Accordingly, Rat-1 fibroblasts that had been exposed to ELF-EMF for 3 or 24 h exhibited a significant increase in dichlorofluorescein-detectable reactive oxygen species, which was blunted by alpha-tocopherol pretreatment. Cells exposed to ELF-EMF and examined as early as 6 h after treatment initiation also exhibited modifications of NF kappa B-related proteins (p65-p50 and I kappa B alpha), which were suggestive of increased formation of p65-p50 or p65-p65 active forms, a process usually attributed to redox reactions. These results suggest that ELF-EMF influence proliferation and DNA damage in both normal and tumor cells through the action of free radical species. This information may be of value for appraising the pathophysiologic consequences of an exposure to ELF-EMF. PMID- 15777848 TI - Clinorotation prevents differentiation of rat myoblastic L6 cells in association with reduced NF-kappa B signaling. AB - In this study, we examined effects of the three-dimensional (3D)-clinorotation, a simulated-model of microgravity, on proliferation/differentiation of rat myoblastic L6 cells. Differentiation of L6 cells into myotubes was significantly disturbed in the 3D-clinorotation culture system, although the 3D-clinorotation had no effect on the proliferation. The 3D-clinorotation also suppressed the expression of myogenesis marker proteins, such as myogenin and myosin heavy chain (MHC), at the mRNA level. In association with this reduced differentiation, we found that the 3D-clinorotation prevented accumulation of ubiquitinated proteins, compared with non-rotation control cells. Based on these findings, we focused on the ubiquitin-dependent degradation of I kappa B, a myogenesis inhibitory protein, to clarify the mechanism of this impaired differentiation. A decline in the amount of I kappa B protein in L6 cells was significantly prevented by the rotation, while the amount of the protein in the non-rotated cells decreased along with the differentiation. Furthermore, the 3D-clinorotation reduced the NF kappaB-binding activity in L6 cells and prevented the ubiquitination of I kappa B proteins in the I kappa B- and ubiquitin-expressing Cos7 cells. Other myogenic regulatory factors, such as deubiquitinases, cyclin E and oxygen, were not associated with the differentiation impaired by the clinorotation. Our present results suggest that simulated microgravity such as the 3D-clinorotation may disturb skeletal muscle cell differentiation, at least in part, by inhibiting the NF-kappa B pathway. PMID- 15777849 TI - Nucleolar localization of hepatic c-Myc: a potential mechanism for c-Myc regulation. AB - The c-myc proto-oncogene encodes a transcription factor that is involved in cell proliferation, growth, differentiation, and apoptosis. Previous studies on the regulation of hepatic c-myc have focused on control of its mRNA expression, which generally correlates with hepatocyte proliferation during both liver development and liver regeneration. However, Western blot analysis showed similar levels of hepatic c-Myc in fetal and adult liver. We therefore went on to examine the abundance and distribution of hepatic c-Myc. Immunofluorescence on adult rat liver cryosections showed that c-Myc was readily detectable, but that it was largely localized to the nucleolus. In contrast, proliferating fetal hepatocytes and adult hepatocytes from regenerating liver showed a diffuse nuclear pattern. Transient transfection of adult hepatocytes with full-length HA-Myc also revealed localization to the nucleolus. Western immunoblotting studies confirmed that immunoreactive c-Myc was present in nucleolar extracts isolated from adult liver. We speculate that the nucleolus may act to sequester c-Myc in quiescent hepatocytes while providing a pool of c-Myc that is readily available to reach its targets in the nucleus. PMID- 15777850 TI - TGF-beta1-mediated activations of c-Src and Rac1 modulate levels of cyclins and p27(Kip1) CDK inhibitor in hepatoma cells replated on fibronectin. AB - Integrin-mediated cell adhesion transduces signals to regulate actin cytoskeleton and cell proliferation. While understanding how integrin signals cross-talk with the TGF-beta1 pathways, we observed lamellipodia formation and cyclin regulation in Hep3B cells, following TGF-beta1 treatment. To answer if integrin signaling via actin organization might regulate cell cycle progression after TGF-beta1 treatment, we analyzed cross-talk between the two receptor-mediated pathways in hepatoma cells on specific ECMs. We found that basal and TGF-beta1-mediated activation of c-Src and Rac1, expression of cyclins E and A, and suppression of p27Kip1 were significant in cells replated on fibronectin, but not in cells on collagen I, indicating a different integrin-mediated cellular response to TGF beta1 treatment. Levels of tyrosine phosphorylation and actin-enriched lamellipodia on fibronectin were also more prominent than in cells on collagen I. Studies using pharmacological inhibitors or transient transfections revealed that the preferential TGF-beta1 effects in cells on fibronectin required c-Src family kinase activity. These observations suggest that a specific cross-talk between TGF-beta1 and fibronectin-binding integrin signal pathways leads to the activation of c-Src/Rac1/actin-organization, leading to changes in cell cycle regulator levels in hepatoma cells. Therefore, this study represents another mechanism to regulate cell cycle regulators when integrin signaling is collaborative with TGF-beta1 pathways. PMID- 15777851 TI - Development of a fluorescent reporter to assess iron regulatory protein activity in living cells. AB - Through the insertion of an iron responsive element (IRE) into a pd2ECFP vector, we demonstrate a noninvasive method for determining alterations in iron regulatory protein (IRP) activity that results in changes in protein translation in living cells. This construct takes advantage of the specifically iron dependent interaction between IRPs that bind IREs on mRNAs to posttranscriptionally regulate protein expression in a manner similar to ferritin production. In this report, we demonstrate, using HEK-293 cells, that an IRE driven fluorescent reporter can be used to observe changes in cellular iron status that are sufficient to alter protein synthesis. When iron availability was decreased, there was less cyan fluorescent protein (CFP) expression, suggesting that IRPs bind to the IRE and block protein translation. Conversely, exposing the cells to iron increased CFP fluorescence. This construct has advantages over traditionally used dyes and existing IRE driven constructs because it can be used to repeatedly study iron-influenced protein production over extended periods of time. The future applications of this construct include investigation of how mutations in cells may impact cellular iron metabolism and how various types of exogenously applied trophic, stress, and therapeutic agents may impact cellular iron metabolism. PMID- 15777852 TI - Regulation of HIV-1 env mRNA translation by Rev protein. AB - We have examined the effect of Rev on the regulation of the expression of RRE containing mRNAs when they were synthesised in the nucleus or directly in the cytoplasm. In the nuclear expression system, Rev enhanced env mRNA transport by about 1.6-fold, while translation of this mRNA was increased more than a 100 fold. These findings indicate that the target of Rev activity is located mainly at the translational level. Synthesis of Env using a recombinant vaccinia virus system, which synthesised env mRNA directly in the cytoplasm, is also enhanced by Rev. Finally, RRE functioning was examined using a luciferase mRNA bearing this element. Rev stimulated the synthesis of Luciferase both when the luc mRNA was made in the nucleus or in cytoplasm. Our results indicate that the effect of Rev on env mRNA transport is low compared with the enhancement of translation of this mRNA. PMID- 15777853 TI - Induction of fusion-competent myoblast-specific gene expression during myogenic differentiation of Drosophila Schneider cells by DNA double-strand breaks or replication inhibition. AB - Differentiation of Drosophila Schneider cells caused by DNA double-strand break (DSB)-inducing topoisomerase II (topo II) inhibitors were attenuated by ICRF-193, a non-DNA-damaging topo II inhibitor. ICRF-193 did not inhibit differentiation induced by neocarzinostatin (NCS), a drug that causes DNA DSBs independent of topo II. Schneider cells differentiated upon treatment with gamma-ray. These results suggest that DNA DSBs induce myogenic differentiation of Schneider cells. We also found DNA replication inhibitors, hydroxyurea (HU), aphidicolin, and ethylmethanesulfonate (EMS) induced myogenic differentiation of Schneider cells. HU-induced differentiation was inhibited upon pretreatment of cells with chemical inhibitors of PP 1/2A, p38 MAPK, JNK, and proteasome. RT-PCR analysis revealed that the expressions of fusion-competent myoblast-specific genes lmd, sns, and del were induced in Schneider cells upon treatment with NCS or HU, whereas expressions of three founder cell-specific genes, duf, ants, and rols, were undetectable. These results indicate that the expression of fusion competent myoblast-specific genes is induced during myogenic differentiation of Drosophila Schneider cells by DNA DSBs or replication inhibition. PMID- 15777855 TI - The burning case of neuropathic pain wording. PMID- 15777854 TI - Acupuncture or acupuncture? PMID- 15777856 TI - Quantitative assessment of experimental pain perception: multiple domains of clinical relevance. PMID- 15777857 TI - Auricular acupuncture for pain relief after total hip arthroplasty - a randomized controlled study. AB - Auricular acupuncture (AA) is known to be effective in treatment of various pain conditions, but still there have been no randomized controlled studies of AA for treatment of acute postoperative pain. Therefore we tested whether AA of specific points is superior to sham acupuncture for complementary analgesia after total hip arthroplasty in a patient-anesthesiologist-evaluator-analyst blinded study. The patients were randomly allocated to receive true AA (lung, shenmen, thalamus and hip points) or sham procedure (4 non-acupuncture points on the auricular helix). Permanent press AA needles were retained in situ 3 days after surgery. Postoperative pain was treated with intravenous piritramide (opioid receptor agonist with analgesic potency of 0.7 compared with morphine) using a patient controlled analgesia (PCA) pump. The time to the first analgesic request, the amount of postoperative piritramide via PCA and pain intensity on a 100-mm visual analogue scale (VAS-100) were used to evaluate postoperative analgesia. Intraoperative anesthetic requirement, incidence of analgesia-related side effects, inflammation parameters and success of patients' blinding were also recorded. Fifty-four patients (29 AA and 25 controls) completed the study. Piritramide requirement during 36 h after surgery in AA group was lower than in control: 37+/-18 vs. 54+/-21 mg; mean+/-SD; P=0.004. Pain intensity on VAS-100 and incidence of analgesia-related side effects were similar in both groups. The differences between the groups as regard patients' opinions concerning success of blinding were not significant. Findings from our study demonstrate that AA could be used to reduce postoperative analgesic requirement. PMID- 15777858 TI - Effects of deliberate control on verbal and facial expressions of pain. AB - The 'facial feedback hypothesis' suggests that inhibiting or exaggerating pain displays produces parallel effects on subjective experience. Research on the regulation of emotional expressions suggests that the act of self-regulation may be detectable in the properties of facial behavior. Both issues were examined in this study. Healthy young volunteers were videotaped while they were exposed to electric shocks varying in intensity. Participants in the Augment group were instructed to exaggerate their facial reactions to the shocks. Participants in the Attenuate group were instructed to inhibit their reactions. Controls simply responded to the shocks. All groups rated the pain of each shock on numeric, sensory and affective scales. In subsequent phases, judges rated the intensity of pain displays for all participants, and facial reactions were measured with the Facial Action Coding System. Results provided no support for the facial feedback hypothesis. Judges' ratings of participants' pain indicated that the augment instructions produced distinct alterations in pain expression. The control and inhibit groups showed linear increases in pain expression with increasing pain intensity, which did not differ significantly. Fine-grained analysis of participants' facial behavior provided evidence that pain augmentation was accompanied by topographic changes in pain expression. Parallels with existing studies, methodological issues and practical implications of the findings are discussed. PMID- 15777860 TI - The anterior cingulate cortex contains distinct areas dissociating external from self-administered painful stimulation: a parametric fMRI study. AB - The anterior cingulate cortex (ACC) has a pivotal role in human pain processing by integrating sensory, executive, attentional, emotional, and motivational components of pain. Cognitive modulation of pain-related ACC activation has been shown by hypnosis, illusion and anticipation. The expectation of a potentially noxious stimulus may not only differ as to when but also how the stimulus is applied. These combined properties led to our hypothesis that ACC is capable of distinguishing external from self-administered noxious tactile stimulation. Thermal contact stimuli with noxious and non-noxious temperatures were self administered or externally applied at the resting right hand in a randomized order. Two additional conditions without any stimulus-eliciting movements served as control conditions to account for the certainty and uncertainty of the impending stimulus. Calculating the differences in the activation pattern between self-administered and externally generated stimuli revealed three distinct areas of activation that graded with perceived stimulus intensity: (i) in the posterior ACC with a linear increase during external but hardly any modulation for the self administered stimulation, (ii) in the midcingulate cortex with activation patterns independent of the mode of application and (iii) in the perigenual ACC with increasing activation during self-administered but decreasing activation during externally applied stimulation. These data support the functional segregation of the human ACC: the posterior ACC may be involved in the prediction of the sensory consequences of pain-related action, the midcingulate cortex in pain intensity coding and the perigenual ACC is related to the onset uncertainty of the impending stimuli. PMID- 15777859 TI - Development and pharmacological characterization of a rat model of osteoarthritis pain. AB - Osteoarthritis (OA) is an age-related joint disease characterized by degeneration of articular cartilage and is associated with chronic pain. Although several experimental models of OA have been employed to investigate the underlying etiologies of the disease, there has been relatively little investigation into development of animal models of OA to study the pain associated with the condition. In the present study, we investigated OA induced by injection of either iodoacetate or papain into the knee joint of rats, and assessed the joint degeneration with radiographic analyses and measured pain behavior using hind limb weight bearing. We found that injection of iodoacetate, but not papain, resulted in a chronic joint degeneration as measured by decreased bone mineral content and bone mineral density, necrosis of articular cartilage and osteophyte formation. These pathological changes were associated with pain that manifested as time- and concentration-dependent alterations in hind limb weight bearing. These alterations in hind limb weight bearing were reversed with morphine, but were not significantly affected by acute administration of either indomethacin or celecoxib. However, administration of 30 mg/kg celecoxib twice daily for 10 days resulted in a significant restoration of hind limb weight bearing. We conclude that the iodoacetate model of OA is a relevant animal model to study pain associated with OA, and can be used to test potential therapeutic agents. PMID- 15777861 TI - The influence of acute anxiety on assessment of nociceptive flexion reflex thresholds in healthy young adults. AB - The nociceptive flexion reflex (NFR) is a polysynaptic withdrawal reflex that occurs in response to painful stimulation. In human studies, NFR responsiveness has been used as a direct measure of nociception as well as an indirect measure of supraspinal modulation of nociceptive transmission. Previous studies have suggested that anxiety may influence NFR responding, and therefore it has been recommended that anxiety be reduced by familiarizing participants with assessment methodology prior to formal NFR assessment. The present study was designed to assess the influence of anxiety on NFR threshold. Using a repeated measures design, 40 men and women completed an NFR threshold assessment twice within session one, and twice again during a second session conducted 24h later. Within each assessment session, state anxiety was measured at the beginning of the session and immediately following each NFR threshold assessment. Results indicated that although anxiety increased in response to NFR threshold assessment and was positively related to subjective pain reports, anxiety was not related to observed NFR threshold levels. These findings suggest that individual differences in anxiety do not significantly affect NFR threshold level determinations under standard testing conditions. PMID- 15777862 TI - Socio-economic differences in the prevalence of acute, chronic and disabling chronic pain among ageing employees. AB - Pain is a strong predictor of reduced work ability and well-being, but there is little information on the prevalence of and socio-economic differences in acute, chronic and disabling chronic pain among employees. A questionnaire survey conducted in 2000-2002 among employees aged 40, 45, 50, 55 and 60 of the City of Helsinki (N=8970, response rate 67%) included socio-demographic and socio economic factors and measures of current pain, pain duration and pain-related disability. Pain was acute when lasting a maximum of 3 months and chronic when persisting for more than 3 months. Disabling chronic pain was determined using the disability subscale of Von Korff's Chronic Pain Grade questionnaire. Acute pain was reported by 15% of women, chronic pain by 29% and disabling chronic pain by 7%. The corresponding figures for men were 12, 24 and 5%. Chronic and disabling chronic pain were more common in older age groups among both genders. Among women, those with secondary or basic education were more likely to report chronic or disabling chronic pain than those with higher education, and semi professionals, routine non-manual employees and manual workers were more likely to report disabling chronic pain than managers. Among men, separated/divorced or widowed men were more likely to report acute pain than married men, and manual workers were more likely to report chronic pain than managers. Chronic pain was relatively common in this population, and those with older age, lower education and occupational class appear to be at excess risk for chronic pain, especially for disabling chronic pain. PMID- 15777863 TI - Cigarette smoking, stress-induced analgesia and pain perception in men and women. AB - This study examined gender differences in smoking-related analgesia and stress induced analgesia (SIA), as a function of pain modality. Forty men (20 smokers, 20 nonsmokers) and 37 women (17 smokers) were tested twice for pain sensitivity to tourniquet ischemia, thermal heat, and cold pressor tests; once following mental stress and once following rest control, counterbalancing order. Cardiovascular and neuroendocrine responses to mental stress were also examined. While expected gender differences in pain sensitivity were observed, women smokers had greater threshold and tolerance times to ischemic pain than women nonsmokers (P<0.05) when pain testing followed rest. Male smokers had greater threshold and tolerance to cold pressor pain than male nonsmokers (P<0.05) after both rest and stress. Only women showed evidence for SIA, since women nonsmokers demonstrated greater ischemic pain threshold and tolerance following mental stress versus rest (P<0.05), and all women reported lower thermal heat pain unpleasantness after stress versus rest (P=0.05). Only nonsmokers showed expected inverse relationships between sympathetic and hypothalamic-pituitary-adrenal (HPA) axis reactivity measures and sensitivity to pain. Smokers showed evidence for blunted HPA-axis function at rest and stress. These results indicate that analgesia related to both being a smoker and stress is influenced by gender and pain modality. The reduced pain perception in smokers and absence of relationships between endogenous pain regulatory mechanisms and pain sensitivity may reflect a maladaptive response to chronic smoking. PMID- 15777864 TI - Disruption of the P2X7 purinoceptor gene abolishes chronic inflammatory and neuropathic pain. AB - The P2X(7) purinoceptor is a ligand-gated cation channel, expressed predominantly by cells of immune origin, with a unique phenotype which includes release of biologically active inflammatory cytokine, interleukin (IL)-1beta following activation, and unique ion channel biophysics observed only in this receptor family. Here we demonstrate that in mice lacking this receptor, inflammatory (in an adjuvant-induced model) and neuropathic (in a partial nerve ligation model) hypersensitivity is completely absent to both mechanical and thermal stimuli, whilst normal nociceptive processing is preserved. The knockout animals were unimpaired in their ability to produce mRNA for pro-IL-1beta, and cytometric analysis of paw and systemic cytokines from knockout and wild-type animals following adjuvant insult suggests a selective effect of the gene deletion on release of IL-1beta and IL-10, with systemic reductions in adjuvant-induced increases in IL-6 and MCP-1. In addition, we show that this receptor is upregulated in human dorsal root ganglia and injured nerves obtained from chronic neuropathic pain patients. We hypothesise that the P2X(7) receptor, via regulation of mature IL-1beta production, plays a common upstream transductional role in the development of pain of neuropathic and inflammatory origin. Drugs which block this target may have the potential to deliver broad-spectrum analgesia. PMID- 15777865 TI - A role for the brainstem in central sensitisation in humans. Evidence from functional magnetic resonance imaging. AB - Animal studies have established a role for the brainstem reticular formation, in particular the rostral ventromedial medulla (RVM), in the development and maintenance of central sensitisation and its clinical manifestation, secondary hyperalgesia. Similar evidence in humans is lacking, as neuroimaging studies have mainly focused on cortical changes. To fully characterise the supraspinal contributions to central sensitisation in humans, we used whole-brain functional magnetic resonance imaging at 3T, to record brain responses to punctate mechanical stimulation in an area of secondary hyperalgesia. We used the heat/capsaicin sensitisation model to induce secondary hyperalgesia on the right lower leg in 12 healthy volunteers. A paired t-test was used to compare activation maps obtained during punctate stimulation of the secondary hyperalgesia area and those recorded during control punctate stimulation (same body site, untreated skin, separate session). The following areas showed significantly increased activation (Z>2.3, corrected P<0.01) during hyperalgesia: contralateral brainstem, cerebellum, bilateral thalamus, contralateral primary and secondary somatosensory cortices, bilateral posterior insula, anterior and posterior cingulate cortices, right middle frontal gyrus and right parietal association cortex. Brainstem activation was localised to two distinct areas of the midbrain reticular formation, in regions consistent with the location of nucleus cuneiformis (NCF) and rostral superior colliculi/periaqueductal gray (SC/PAG). The PAG and the NCF are the major sources of input to the RVM, and therefore in an ideal position to modulate its output. These results suggest that structures in the mesencephalic reticular formation, possibly the NCF and PAG, are involved in central sensitisation in humans. PMID- 15777866 TI - Prognostic factors for poor recovery in acute whiplash patients. AB - The objective of our prospective inception cohort study was to identify prognostic factors for poor recovery in patients with whiplash-associated disorders grade 1 or 2 who still had neck pain and accompanying complaints 2 weeks after the accident. The study was carried out in a primary health care setting in The Netherlands and included 125 patients. The primary outcome measure was functional recovery defined in terms of neck pain intensity or work disability without medication use. The secondary outcome measures included neck pain intensity, work disability and sick leave. The outcomes were assessed at 4, 12 and 52 weeks after the accident. Prognostic factors were identified by logistic regression analyses. One year after the injury, 64% of the patients were recovered. Factors related to poor recovery were female gender, a low level of education, high initial neck pain, more severe disability, higher levels of somatisation and sleep difficulties. Neck pain intensity and work disability proved to be the most consistent predictors for poor recovery. The accuracy of the predictions of the prognostic models was high, meaning that the models adequately distinguished patients with poor recovery from those regarded as recovered. These findings add to the growing body of evidence, indicating that socio-demographic, physical and psychological factors affect short- and long-term outcome after whiplash injury. Our findings also indicate that care providers can easily identify patients at risk for poor recovery with a visual analogue scale for initial pain intensity and work-related activities. PMID- 15777867 TI - Characterization of long-term potentiation of primary afferent transmission at trigeminal synapses of juvenile rats: essential role of subtype 5 metabotropic glutamate receptors. AB - Recent work has demonstrated that a brief high-frequency conditioning stimulation to the primary afferent nerve fibers can induce a long-term potentiation (LTP) of synaptic transmission in neurons in the superficial layer of the trigeminal caudal nucleus; however, the cellular and molecular mechanisms underlying this synaptic potentiation remain unclear. Using both extracellular field potential and whole-cell patch-clamp recordings in brainstem parasagital slices of juvenile rat with the mandibular nerve attached, we show here that the induction of trigeminal primary afferent LTP: (1) does not require the activation of ionotropic glutamate receptors; (2) is dependent on extracellular Ca(2+) and the release of Ca(2+) from intracellular stores; (3) is specifically prevented by the metabotropic glutamate receptor subtype 5 (mGluR5) antagonist 2-methyl-6 (phenylethynyl)pyridine but not the mGluR1 antagonist LY367385, group II mGluR antagonist LY341495 or group III mGluR antagonist MAP4; (4) is mimicked by the bath-applied group I mGluR agonist (S)-3,5-dihydroxyphenylglycine and mGluR5 agonist (RS)-2-chloro-5-hydroxyphenylglycine; (5) requires the activation of phospholipase C (PLC) and protein kinase C (PKC); and (6) is concomitantly with a decrease in paired-pulse depression. These results demonstrate that the activation of mGluR5 and in turn triggering a PLC/PKC-dependent signaling cascade may contribute to the induction of LTP of primary afferent synaptic transmission in the superficial layer of trigeminal caudal nucleus of juvenile rats. This may be relevant to the processing of nociceptive information. PMID- 15777868 TI - New evidence that both T-type calcium channels and GABAA channels are responsible for the potent peripheral analgesic effects of 5alpha-reduced neuroactive steroids. AB - Neurosteroids are potent blockers of neuronal low-voltage activated (T-type) Ca(2+) channels and potentiators of GABA(A) ligand-gated channels, but their effects in peripheral pain pathways have not been studied previously. To investigate potential analgesic effects and the ion channels involved, we tested the ability of locally injected 5alpha-reduced neurosteroids to modulate peripheral thermal nociception to radiant heat in adult rats in vivo and to modulate GABA(A) and T-type Ca(2+) channels in vitro. The steroid anesthetic alphaxalone (ALPX), the endogenous neurosteroid allopregnanolone (3alpha5alphaP), and a related compound ((3alpha,5alpha,17beta)-3-hydroxyandrostane-17 carbonitrile, (ACN)), induced potent, dose-dependent, enantioselective anti nociception in vivo and modulation of both T-type Ca(2+) currents and GABA(A) mediated currents in vitro. Analgesic effects of ALPX were incompletely antagonized by co-injections of the GABA(A) receptor antagonist bicuculline. The neurosteroid analogue ((3alpha,5alpha)-3-hydroxy-13,24-cyclo-18,21-dinorchol-22 en-24-ol (CDNC24), a compound with GABAergic but not T-type activity, was not analgesic. However, (3beta,5alpha,17beta)-17-hydroxyestrane-3-carbonitrile (ECN)), which has effects on T-type channels but not on GABA(A) receptors, also induced potent enantioselective peripheral anti-nociception. ECN increased pain thresholds less than ALPX, 3alpha5alphaP and ACN. However, when an ineffective dose of CDNC24 was combined with ECN, anti-nociceptive activity was greatly enhanced, and this effect was bicuculline-sensitive. These results strongly suggest that GABA(A) channels do not contribute to baseline pain transmission, but they can enhance anti-nociception mediated by blockade of T-type Ca(2+) channels. In conclusion, we demonstrate that potent peripheral analgesia induced by 5alpha-reduced neurosteroid is mediated in part by effects on T-type Ca(2+) channels. Our results also reveal a role of GABA-gated ion channels in peripheral nociceptive signaling. PMID- 15777869 TI - Does neonatal surgery lead to increased pain sensitivity in later childhood? AB - Does pain or tissue damage in early life lead to hyperalgesia persisting into childhood? We performed a cross-sectional study in 164 infants to investigate whether major surgery within the first 3 months of life increases pain sensitivity to subsequent surgery and to elucidate whether subsequent surgery in the same dermatome or in a different dermatome leads to differences in pain sensitivity. All infants received standard intraoperative and postoperative pain management, with rescue analgesia guided by a treatment algorithm. Differences in pain sensitivity during surgery were assessed by the intraoperative fentanyl intake and by (nor)epinephrine plasma concentrations. Differences in postoperative pain sensitivity were assessed by the observational pain measures COMFORT and VAS, and by morphine intake and (nor)epinephrine plasma concentrations. Infants previously operated upon in the same dermatome needed more intraoperative fentanyl, had higher COMFORT and VAS scores, had greater (nor)epinephrine plasma concentrations, and needed also more morphine than did infants with no prior surgery. In contrast, infants who previously underwent surgery in another dermatome had only significant higher postoperative analgesic requirements and norepinephrine plasma concentrations in comparison with infants with no prior surgery. These preliminary differences may indicate the occurrence of spinal and supraspinal changes following neonatal surgery. We conclude that the long-term consequences of surgery in early infancy are greater in areas of prior tissue damage and that these effects may portend limited clinical but important neurobiological differences. PMID- 15777870 TI - Analgesic effects of dietary caloric restriction in adult mice. AB - Nociception was studied in male mice, mostly of the C57BL/6 strain, during continuous or prolonged restriction of caloric intake (60% of ad-libitum) from midlife to senescence (up to 105 weeks). Restricted mice showed fewer licking or biting responses 20-60 min after hind paw injection of 5% formalin at 46 and 70 weeks, but not at 93 weeks. Also, they showed longer response latencies around 46 weeks of age in the 52 degrees C hot-plate test, which partial tail amputation failed to affect, although it did produce at least 2 weeks of chronic neuropathic hypersensitivity in ad libitum controls. Injection of collagen subcutaneously at 36-42 weeks led to chronic hyperalgesia in the DBA/1 but not the C57BL/6 strain, measured weekly by the barely nociceptive 50 degrees C hot-plate test to minimize damage. This collagen-induced arthritic hyperalgesia was then gradually and reversibly blocked during 9-15 weeks of caloric restriction starting at 53-58 weeks. In longitudinal trials on normal mice, performed every 2-4 weeks between 42 and 105 weeks with the 50 degrees C hot-plate, caloric restriction led to altered latencies (higher relative to controls) only in the last 10-20 weeks, perhaps because it delayed the onset of age-related peripheral neuropathies. In conclusion, long-term caloric restriction leads to significant hypoalgesia in pre senescent mice subjected to above-threshold pain of widely different durations, the effect disappearing at later ages unless spontaneous neuropathies become influential. A reduction in cumulative food intake thus appears to generate antinociceptive signals in adult male mice, perhaps serving specifically to promote riskier behavior during prolonged food shortages. PMID- 15777871 TI - Race and socioeconomic differences in post-settlement outcomes for African American and Caucasian Workers' Compensation claimants with low back injuries. AB - The purpose of this study was to predict post-settlement pain intensity, psychological distress, disability, and financial struggle among African American (n=580) and non-Hispanic Caucasian (n=892) Workers' Compensation claimants with single incident low back injury. The study was a population-based telephone survey conducted in three population centers in Missouri. Post-settlement outcomes were predicted from claimant demographics (race, age, gender); socioeconomic status (SES); diagnosis and legal representation; and Workers' Compensation resolution variables (treatment costs, temporary disability status, disability rating, settlement costs). Simultaneous-entry, hierarchical multiple linear regression analyses indicated that African American race and lower SES predicted higher levels of post-settlement pain intensity, psychological distress (general mental health, pain-related catastrophizing), disability (pain-related role interference), and financial struggle, independent of age, gender, diagnosis, legal representation, and Workers' Compensation resolution variables. The results suggest that African American race and lower SES-relative to Caucasian race and higher SES-are risk factors for poor outcomes after occupational low back injury. Mechanisms to explain these associations are discussed, including patient-level, provider-level, legal, and Workers' Compensation system-level factors. PMID- 15777872 TI - Sensory function and quality of life in patients with multiple sclerosis and pain. AB - Central neuropathic pain is well known in multiple sclerosis (MS), but the underlying mechanisms are unclear. In the present study we studied sensory function in MS patients with pain, MS patients without pain and healthy subjects in order to clarify the role of sensory abnormalities in pain. Fifty MS patients with pain were randomly recruited from a previous epidemiological MS study in Aarhus County, Denmark. Age and gender stratified MS patients without pain (N=50) and healthy subjects (N=50) served as controls. Patients with pain underwent a structured pain interview. Sensory function was examined by bedside and quantitative sensory testing. Quality of life was assessed using the health related quality of life questionnaire, SF-36. Patients with pain had lower pressure pain threshold than pain-free patients (260 kPa vs. 322 (median), P=0.02) otherwise quantitative sensory testing was similar. Pain patients more frequently had cold allodynia (9/50 vs. 0/50, P=0.003) and abnormal temporal summation (10/48 vs. 3/49, P=0.03). Fifty-eight percent had central pain. Central pain patients did not differ from musculoskeletal pain patients in quantitative sensory testing, but allodynia was more common in MS patients with central pain. Pain patients scored lower in all dimensions of SF-36 compared with pain-free patients and healthy subjects. The results suggest that pain in MS is central in more than half of the patients and is associated with mechanical or thermal hyperalgesia. PMID- 15777873 TI - Spinal 5-HT1A receptors differentially influence nociceptive processing according to the nature of the noxious stimulus in rats: effect of WAY-100635 on the antinociceptive activities of paracetamol, venlafaxine and 5-HT. AB - The regulation of nociceptive processing by 5-HT at the spinal level is intricate since the neurotransmitter has been implicated in both pro and antinociception. The aim of our study was to investigate, according to the nature of the noxious stimulus, how the blockade of spinal 5-HT(1A) receptors could influence the antinociceptive actions of exogenous 5-HT as well as two analgesics involving endogenous 5-HT, paracetamol and venlafaxine. Rats were submitted either to the formalin test (tonic pain) or the paw pressure test (acute pain). WAY-100635 (40 microg/rat, i.t.), a selective 5-HT(1A) receptor antagonist, had no intrinsic action in either test. However, in the formalin test, it blocked the antinociceptive action of 5-HT (50 microg/rat, i.t.) and paracetamol (300 mg/kg, i.v.) in both phases of biting/licking behaviour and that of venlafaxine (2.5 mg/kg, s.c.) in the late phase only. In the paw pressure test, the combination of sub-effective doses of 5-HT (0.01 microg/rat, i.t.), paracetamol (50 mg/kg, i.v.) or venlafaxine (20 mg/kg, s.c.) with WAY-100635 led to a significant antinociceptive effect, which seems to depend on the reinforcement of the activity of inhibitory GABAergic interneurones. In conclusion, both direct stimulation of the spinal 5-HT(1A) receptors by 5-HT, and indirect stimulation using paracetamol or venlafaxine can differently influence pain transmission. We propose that the nature of the applied nociceptive stimulus would be responsible for the dual effect of the 5-HT(1A) receptors rather than the hyperalgesic state or the supraspinal integration of the pain message. PMID- 15777874 TI - Do health care providers' attitudes towards back pain predict their treatment recommendations? Differential predictive validity of implicit and explicit attitude measures. AB - The current study aimed to measure the differential predictive value of implicit and explicit attitude measures on treatment behaviour of health care providers. Thirty-six physiotherapy students completed a measure of explicit treatment attitude (Pain Attitudes And Beliefs Scale For Physiotherapists-PABS-PT) and a measure of implicit treatment attitude (Extrinsic Affective Simon Task-EAST). Furthermore, they gave treatment recommendations for a patient simulating back pain on three video scenes. The implicit and explicit measures of attitudes were only weakly related to each other. However, both were differentially related to treatment recommendations. The implications of the differential predictive value of implicit and explicit attitude measures for treatment behaviour are discussed. PMID- 15777875 TI - Spinal administration of MK-801 and NBQX demonstrates NMDA-independent dorsal horn sensitization in incisional pain. AB - Surgery commonly causes pain and neural plasticity that are unique compared to other persistent pain problems. To more precisely study central sensitization and plasticity, we examined the role of ionotropic EAA receptors in dorsal horn neuron sensitization early after incision. Sensitization, in the form of increased background activity, increased mechanosensitivity or pinch receptive field expansion, was induced by plantar incision 1 h later in 30 neurons. (+)-5 Methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine (MK-801) or 1 mM 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo[f]quinoxaline-7-sulfonamide (NBQX) was administered through a microdialysis fiber to block NMDA and nonNMDA EAA receptors, respectively. Dorsal horn neuron sensitization was reexamined 1 h later. Spinal administration of NBQX blocked AMPA-induced excitation but did not affect excitation by NMDA. NBQX decreased background activity in the neurons that developed sustained increased activity after incision. The median decrease caused by NBQX was from 2.3 to 0.0 imp/s. Spinal administration of 5 mM MK-801 blocked NMDA-induced excitation but did not affect excitation by AMPA. The median change (from 2.6 to 1.1 imp/s) in background activity increased by incision was not significantly affected by MK-801. The responses to mechanical stimuli were enhanced after incision in wide dynamic range (WDR) neurons. NBQX eliminated these responses but MK-801 had no effect. The pinch receptive field (RF) expansion into uninjured areas of the paw and hindquarters occurred after incision. Only 1 of 13 neurons exhibited RF expansion after spinal NBQX administration; 9 of 12 neurons had RF expansion remaining after MK-801. Thus, nonNMDA receptors are critical and NMDA-independent factors influence the increased responsiveness of dorsal horn neurons that occur early after incision. PMID- 15777876 TI - Acupuncture analgesia during surgery: a systematic review. AB - The aim of this systematic review is to assess the effectiveness of acupuncture as an adjunctive analgesic method to standard anaesthetic procedures for surgery and to determine whether acupuncture has any analgesic-sparing effect. Electronic literature searches for randomised clinical trials (RCTs) of acupuncture during surgery were performed in seven electronic databases. No language restrictions were imposed. All included studies were rated according to their methodological quality and validity. As the studies were clinically heterogeneous, no meta analyses were performed. The evidence was classified according to four levels: strong, moderate, limited, or inconclusive. Nineteen RCTs were identified. Seven of them suggested that acupuncture is efficacious. Of nine high-quality RCTs, two studies had positive outcomes. There was no significant association between study quality and direction of outcome. One of eight high-validity trials reported a positive outcome and there was a significant relationship between validity and direction of outcome. The evidence that acupuncture is more effective than no acupuncture as an adjunct to standard anaesthetic procedures is therefore inconclusive. Strong evidence exists that real acupuncture is not significantly different from placebo acupuncture. For an analgesic-sparing effect of acupuncture, evidence remains inconclusive. In conclusion, this review does not support the use of acupuncture as an adjunct to standard anaesthetic procedures during surgery. PMID- 15777877 TI - Impaired self-perception of the hand in complex regional pain syndrome (CRPS) [S. Forderreuther, U. Sailer, A. Straube, Pain 2004; 110:756-761]. PMID- 15777878 TI - Comment on "Pain relief: a universal right" by Cousins, Brennan and Carr. PAIN Vol 112/1-2, pp. 1-4. PMID- 15777880 TI - Assessment of pain in laboratory animals: a comment on Mogil and Crager (2004). PMID- 15777883 TI - Cognitive modulation of pain-related brain responses. Comments on Seminowicz et al. (Pain 2004;112:48-58). PMID- 15777885 TI - Extremity soft tissue sarcoma: controversial management issues. AB - Unlike common malignancies, such as breast and colorectal carcinoma, where treatment modalities can be investigated with large prospective randomized trials, such an endeavor has been hampered with soft tissue sarcomas (STS) due to its rarity. In absence of such randomized clinical trials, controversy exists with regards to numerous clinically relevant questions and clinicians are left with single institutional experiences gathered either in a retrospective or prospective fashion. Some of these frequently encountered issues in the management of STS include (1) whether poorly executed biopsies affect outcome? (2) Do all unplanned excisions require re-excisions? (3) Is MRI a superior imaging modality? (4) Whether radiation should be provided pre- or post operatively? (5) Does extent of surgical margin influence local control? (6) Is adjuvant radiation therapy necessary for stage IIB STS? (7) Does adjuvant chemotherapy influence local control? (8) Does local recurrence influence survival? We will address these topics in this review. PMID- 15777886 TI - Hepatoid malignancy of unknown origin--a diagnostic conundrum: review of literature and case report of collision with adenocarcinoma. AB - Hepatoid carcinomas are a group of neoplasms with features resembling hepatocellular carcinomas. Although rare, more cases have been noted to arise from various organs within the last decade. Differentiating these tumors is not only a challenge but also critical, since treatment modalities and operative strategies are dependent upon the exact nature of the hepatoid cancer. Based on a review of literature, we discuss the guidelines for differentiating these tumors and utilize these criteria to differentiate these tumors irrespective of their primary tissue of origin. We also describe an unusual case of hepatoid variant of primary peritoneal yolk sac tumor presenting with extensive carcinomatosis and as a collision with two synchronous primary colonic adenocarcinomas, neither of which has been reported to our knowledge to date, thereby falsely mimicking metastatic dedifferentiated colonic adenocarcinoma. CASE DESCRIPTION: A septuagenarian female presented with weight loss, chronic anemia and ascites associated with bloating. Her past history was significant for COPD, hysterectomy and a bilateral oophorectomy 40 years previously. A work up which included computed tomography (CT) and contrast enema revealed significant ascites with multiple peritoneal nodules causing ascending colonic obstruction. Celiotomy revealed the entire peritoneal surface to be studded with nodules, with the largest measuring 6.0 cm. Intraoperative ultrasound, prompted by the frozen section of nodules resembling liver tissue, revealed no intrahepatic nodules. Right colectomy, omentectomy and tumor debulking were performed. The postoperative course was uneventful except for ascitic leak that spontaneously resolved. DISCUSSION: The peritoneal nodules consisted of malignant cells arranged in cords that resembled liver tissue. In the absence of a primary identifiable liver disease, this is consistent with either hepatoid variant of primary yolk sac tumor or hepatoid carcinoma arising from the peritoneum. The right colectomy specimen revealed two mucosal ulcers consistent with colonic adenocarcinoma abutting two large tumor nodules on the serosal surface. Although grossly appearing to be contiguous, low power examination showed that the serosal nodule and the mucosal lesion were entirely different, and separated by a definite fibrous band. Immunohistochemical stains and patterns were used to differentiate the type of tumor. This article also discusses the criteria used to differentiate hepatoid yolk sac tumors (hepatoid-YSTs), hepatoid carcinomas and metastatic hepatocellular carcinomas and the phenomenon of opisthoplasia observed in certain malignancies. The difficulties encountered in identifying hepatoid carcinomas and current modalities used to differentiate these tumors are highlighted. The phenomenon of opisthoplasia and the challenges posed by this phenomenon in certain metastatic lesions are reviewed and explanations for the possible origins of hepatoid tumors are considered. CONCLUSION: The most likely explanation in our case is that the hepatoid tumor either originated from microscopic remnant ovarian tissue left behind or primarily from the peritoneum. With no evidence of yolk sac component within the colonic tumor or in the draining lymphatics, this essentially excludes the commonly observed metastatic dedifferentiation (opisthoplasia) of adenocarcinoma to primitive forms (also known as combination tumors). Based on analysis of various factors, including tumor behavior and response to chemotherapy, we conclude that our case was a hepatoid variant of yolk sac tumor presenting in an elderly woman with carcinomatosis. This unusual presentation of two entirely different primary malignancies in close proximity is defined as "collision tumor". This is the first reported case of collision tumors involving dual colonic and primary peritoneal hepatoid-YST. Identifying the exact type still remains to be the most challenging aspect in the diagnosis of hepatoid tumors. PMID- 15777887 TI - New therapeutic approaches for early stage non-small cell lung cancer. AB - Surgical resection remains the mainstay of therapy for early stage non-small cell lung cancer (NSCLC). Unfortunately, many patients present with advanced stage disease, and many with resectable early stage disease are unable to tolerate pulmonary resection because of compromised cardiopulmonary function. This article reviews the standard and some alternative therapies that are being introduced into clinical practice for early stage NSCLC. New therapies such as sublobar resection with brachytherapy, radiofrequency ablation and stereotactic radiosurgery offer some hope for those patients who are deemed poor candidates for curative resection. PMID- 15777888 TI - Long-term management of patients after potentially curative treatment of extremity soft tissue sarcoma: practice patterns of members of the Society of Surgical Oncology. AB - The optimal strategy for follow-up of extremity soft tissue sarcoma patients after primary treatment with curative intent is not known. The 1592 members of the Society of Surgical Oncology (SSO) were sent a survey to determine their practices. Office visits and chest X-rays were the most frequently performed items for all years. Complete blood count and liver function tests were the most commonly ordered blood tests, but many respondents did not order any blood tests routinely. Imaging studies of the extremities were recommended for most patients during the first postoperative year. Postoperative surveillance strategies after sarcoma treatment vary appreciably among surgeons. Surgeon age, tumor size, tumor grade, and geographic location of the surgeon account for only a modest amount of the variation, suggesting that lack of evidence supporting any particular strategy is the factor responsible for most of the variation. Efforts to provide high-quality evidence on this topic are warranted. PMID- 15777889 TI - Radiofrequency ablation for malignant liver tumor. AB - Radiofrequency ablation (RFA) has been widely practiced to treat unresectable malignant liver tumors. It has the merits of localized tumor ablation and preservation of maximal normal liver parenchyma. In recent years, there has been a tremendous expansion in the application of RFA for patients with malignant liver tumors. However, the therapeutic effect of this local ablation treatment needs to be balanced against its risks and possible local failure. This review focuses on the current status of RFA for malignant liver tumors, with special attention to the indication, approaches, complications, survival benefits, combination therapies, and comparison with other treatment modalities. Although the results of most clinical studies of RFA seem favorable, the associated risks and tumor recurrence should not be underestimated. Careful patient selection, meticulous RFA techniques, and prompt treatment of residual and recurrent tumors are necessary to ensure a better outcome after RFA. Until recently, there has been no strong evidence showing that RFA can replace any other treatment modalities in the management of liver tumors. Nonetheless, more convincing evidence by randomized trials is required for the establishment of a treatment protocol of RFA for patients with malignant liver tumors. PMID- 15777891 TI - Oral versus high dose parenteral iron supplementation in pregnancy. AB - OBJECTIVE: To compare the effect of daily oral iron supplementation with two injections of high dose parenteral iron. METHOD: A total of 220 pregnant women with a singleton pregnancy and hemoglobin between 8 to 11 g% at 16-24 weeks gestation were identified and randomly divided into two groups. Group A was started on daily oral iron therapy of 100 mg of elemental iron. Group B was given 250 mg of iron sorbitol intramuscularly and repeated at an interval of 4-6 weeks. Blood indices were evaluated at the beginning of study and at 36 weeks to see the effect after iron supplementation in the two groups. The data were analyzed using SPSS software, version 10.1. RESULTS: Definitive and comparable improvement in hemoglobin and all the blood indices (hematocrit, MCH, MCHC, MCV, Serum iron and TIBC) was observed. The absolute change in hemoglobin and hematocrit was 1.18+/ 0.68 g% and 4.02+/-2.59% in oral group, 1.34+/-0.77 g% and 4.93+/-3.65% in parenteral group, respectively. Serum ferritin showed statistically significant absolute rise (10.43+/-7.92 microg/dl) after parenteral iron supplementation as compared to oral iron supplementation (9.76+/-4.78 microg/dl). Obstetric outcome was comparable in two groups. CONCLUSION: Two treatment regimens are biologically equivalent in terms of hematological response. Two high doses of intramuscular iron can be a good substitute to meet iron requirement in pregnancy. PMID- 15777892 TI - Diagnosis of human cytomegalovirus intrauterine infection using fetal cells from maternal blood. AB - OBJECTIVE: The sensitivity and specificity for the noninvasive prenatal diagnosis of human cytomegalovirus intrauterine infection were estimated by using isolating single fetal cells from maternal peripheral blood. METHODS: Micromanipulation techniques were employed to isolate single fetal nucleated erythroblasts from 273 maternal blood samples. SRY gene and HCMV-DNA in single fetal cells were detected by multiple primed in situ labeling (PRINS) from 76 HCMV-DNA positive samples of maternal peripheral blood. 273 samples of maternal peripheral blood were tested for SRY gene and HCMV-DNA in single fetal cells by primed extension preamplification (PEP) and polymerase chain reaction (PCR). RESULTS: The detection rate of fetal cells from maternal blood was 100% with micromanipulation techniques. The sensitivity of PRINS for SRY gene detection was 97.56% and its specificity was 100%. The sensitivity and specificity of PEP and PCR for SRY gene detection were 97.39% and 99.17%, respectively. The sensitivity of PRINS for HCMV DNA detection was 92.68% and the specificity was 100%. The sensitivity and specificity of PEP and PCR for HCMV-DNA detection were 95.12%and 100%, respectively. CONCLUSION: The technique for noninvasive prenatal detection of intrauterine infection of HCMV using single fetal cells from maternal peripheral blood by using PRINS and PEP and PCR is more reliable than the CMV-DNA detection in peripheral maternal blood, amniocentesis or percutaneous umbilical blood sampling. PMID- 15777893 TI - Lamellar body count as a predictor of neonatal lung maturity in high-risk pregnancies. AB - OBJECTIVES: To compare the usefulness of a lamellar body count, a fluorescence polarization assay, and the foam stability index for predicting neonatal lung maturity in high-risk pregnancies. SETTING: This study was conducted at the Department of Obstetrics and Gynecology and the Department of Pediatrics, Assiut University Hospital. DESIGN: A prospective clinical trail. SUBJECTS AND METHODS: This study was performed after recruiting 73 pregnant women, 52 with high-risk pregnancies (25 had diabetes and 27 had premature labor) and 21 with a healthy full-term singleton pregnancy as controls. All women were delivered in the Department of Obstetrics and Gynecology of Assiut University Hospital. The newborns with respiratory distress syndrome (RDS) were admitted in the neonatal intensive care unit of the Department of Pediatrics. Amniotic fluid specimens were obtained near delivery. Apgar score, vital signs, anthropometric data, and complete clinical examination results were available for all newborns, and particular emphasis was placed on signs of RDS. RESULTS: The incidence of RDS was 44.2% in the newborns of women who had experienced a high-risk pregnancy (of these, 82.6% were born preterm and 17.4% to diabetic mothers). We found that a lamellar body count is a good screening test for predicting neonatal lung maturity. It is as good as the fetal lung maturity assay by fluorescence polarization in some respects and better in others; moreover, it is better than the foam stability index test in all respects. A lamellar body count with cutoffs of 41 x 10(3)/microL and 18 x 10(3)/microL was a good predictor of low and high risks of RDS in newborns. Values between 19 x 10(3)/microL and 40 x 10(3)/microL were the best to predict an intermediate risk of RDS. CONCLUSION: Lamellar body count is a good screening test for predicting the degree of neonatal lung maturity. PMID- 15777894 TI - Helicobacter pylori infection and platelet counts during pregnancy. AB - OBJECTIVE: To investigate whether there is any correlation between Helicobacter pylori infection and platelet counts during pregnancy. METHODS: One hundred and twenty pregnant women without any complications were evaluated. Platelet counts were determined during pregnancy and postpartum. H. pylori infection was assessed by the detection of serum antibodies at the 3rd day postpartum. Statistically significant differences of platelet count between H. pylori-positive and negative patients were determined. RESULTS: H. pylori infection was found in 29 of 120 (24.2%). Platelet counts in the 3rd trimester were 22.7+/-4.7 x 10(3)/microl in H. pylori-positive group and 22.8+/-5.6 x 10(3)/microl in negative group (p=0.98). Platelet count in the 3rd trimester in positive and negative patients were 93+/-17% and 94+/-20%, respectively, of that of the 1st trimester (p=0.92). The incidence of hyperemesis gravidarum in both groups was not significant (p=0.28). CONCLUSION: A correlation between H. pylori infection and thrombocytopenia during pregnancy was not found in this study. PMID- 15777895 TI - Ultrasound imaging to evaluate Creatsas vaginoplasty. AB - OBJECTIVE: To present a new imaging technique for the evaluation of the functional effectiveness of Creatsas vaginoplasty. METHOD: Eighteen women with Mayer-Rokitansky-Kuster-Hauser syndrome underwent a transabdominal and transperineal ultrasound examination 4 weeks and 6 months after the surgical creation of a neovagina, and then yearly, with a condom filled with water as a distending and imaging medium. The functional length and width of the neovagina and its axis deviation were assessed, as well as the postoperative quality of the women's sexual life. RESULTS: A functional vagina 10 to 12 cm in length and 4 to 5 cm in width was observed; its axis deviation, which was similar to the anatomic deviation, was easily determined; and 94.5% of the women reported a satisfactory while 5.5% reported an adequate sexual life. CONCLUSION: This new imaging technique is a simple and effective alternative for the evaluation of the postoperative effectiveness of colpopoiesis. PMID- 15777896 TI - Emergency craniotomy in patients with intracranial metastatic gestational trophoblastic tumor. AB - OBJECTIVE: To study the diagnosis, management and prognosis of intracranial metastases of gestational trophoblastic tumor (GTT) patients who had emergency craniotomy. METHODS: Sixty three intracranial metastases of GTT patients were hospitalized in Peking Union Medical College Hospital from 1985 to 2004. Thirteen of them had craniotomy that were retrospectively analyzed and all of them had a feature of raised intracranial pressure followed by intracranial hemorrhagic mass. RESULTS: Of the thirteen patients who had undergone surgery combined with multiagent chemotherapy, seven got complete remission (CR), five got partial remission (PR), and one died of progress of the disease (PD). There were three patients who died (23.1%) during or after therapy. CONCLUSIONS: Emergency craniotomy has a small but important role to play in selected patients with brain metastases who display rapidly deteriorating signs. An undetermined intracranial hemorrhage in young female patients of a reproductive age group should raise the suspicion of cerebral metastatic GTT. PMID- 15777897 TI - Clinical aspects and prognosis of pelvic recurrence of cervical carcinoma. AB - OBJECTIVE: To identify which patients with locally recurrent cervical carcinoma are potentially curable. METHOD: A total of 664 stage IB-IVA patients were examined following surgery or radiotherapy. RESULT: Among the 664 patients, 193 (29%) developed recurrence. Sixty-seven (35%) of these recurrences were located in the pelvis alone. Among these 67 recurrences, 24 (35%) were central recurrences and the remaining 43 (65%) were pelvic side-wall recurrences. Of the 24 patients with central recurrences, 8 were salvaged. Of these 8 patients, 3 underwent pelvic exenteration, and 5 received optimal radiotherapy. The recurrent tumor in these 5 survivors who received radiotherapy had consisted of a small (<2 cm) tumor. All 43 patients with pelvic wall recurrence developed progressive disease. CONCLUSION: The following patients are potentially curable: patients with a resectable, centrally located tumor who are candidates for pelvic exenteration, and patients with a small central recurrence for whom complete radiation therapy is feasible. PMID- 15777898 TI - Maternal and umbilical soluble Fas concentration in normotensive and preeclamptic pregnancies. PMID- 15777899 TI - Subclinical thrombocytopenia and preeclampsia. PMID- 15777900 TI - Using the kanga to measure postpartum blood loss. PMID- 15777901 TI - Maternal serum ceruloplasmin in preeclampsia. PMID- 15777902 TI - Marginal involvement after large loop excision of transformation zone in patients with cervical cancerous lesion. PMID- 15777903 TI - Neurodevelopmental and cognitive assessment of 6-year-old children born growth restricted. PMID- 15777904 TI - Single dose of 1.5 mg Levonorgestrel for emergency contraception. PMID- 15777905 TI - Abdominal wall endometriomas. PMID- 15777906 TI - Idiopathic chronic uterine inversion in a nulligravida. PMID- 15777907 TI - Optical diagnosis of cervical dysplasia. PMID- 15777908 TI - Daidzein-rich isoflavone-aglycones for menopausal symptoms. PMID- 15777909 TI - Cervical softening with vaginal misoprostol before intrauterine device insertion. PMID- 15777910 TI - Attitudes about medical abortion among Indian women. PMID- 15777911 TI - Isolation and characterisation of a beta tubulin isotype 2 gene from two species of cyathostomin. AB - This study describes the isolation and characterisation of beta tubulin isotype 2 cDNA sequences from two common species of cyathostomin, Cylicocyclus nassatus and Cyathostomum catinatum. The full-length cDNAs for these species were 1709 and 1753 bp in length, respectively, including 1350 bp of sequence inferred to encode 450 amino acids of peptide sequence. They had greatest identity with previously characterised isotype 2 sequences from Trichostrongylus colubriformis, Cooperia oncophora and Haemonchus contortus (96% for C. nassatus and 95% for C. catinatum), and grouped together with these sequences in phylogenetic analysis. Both cyathostomin beta tubulin isotype 2 sequences contained the isotype-specific carboxyl terminal region described previously in other nematode species. Alignment with beta tubulin isotype 1 proteins from other trichostrongyloids, indicated 95 and 94% identity for the isotype 2 sequences of C. nassatus and C. catinatum, respectively. This comparison revealed 14 isotype-specific amino acid substitutions. Also, 2605 bp of beta tubulin isotype 2 genomic DNA sequence were isolated from C. nassatus. Comparison with the previously published isotype 1 gene of C. nassatus indicated differences in genomic organisation between the two isotypes. Reverse transcriptase (RT)-polymerase chain reaction analysis revealed constitutive temporal expression of beta tubulin isotype 1, whilst isotype 2 appeared to be developmentally expressed, with transcripts detected only in RNA derived from adult parasites. PMID- 15777912 TI - The Trypanosoma brucei La protein is a candidate poly(U) shield that impacts spliced leader RNA maturation and tRNA intron removal. AB - By virtue of its preferential binding to poly(U) tails on small RNA precursors and nuclear localisation motif, the La protein has been implicated for a role in the stabilisation and nuclear retention of processing intermediates for a variety of small RNAs in eukaryotic cells. As the universal substrate for trans-splicing, the spliced leader RNA is transcribed as a precursor with just such a tail. La protein was targeted for selective knockdown by inducible RNA interference in Trypanosoma brucei. Of three RNA interference strategies employed, a p2T7-177 vector was the most effective in reducing both the La mRNA as well as the protein itself from induced cells. In the relative absence of La protein T. brucei cells were not viable, in contrast to La gene knockouts in yeast. A variety of potential small RNA substrates were examined under induction, including spliced leader RNA, spliced leader associated RNA, the U1, U2, U4, and U6 small nuclear RNAs, 5S ribosomal RNA, U3 small nucleolar RNA, and tRNATyr. None of these molecules showed significant variance in size or abundance in their mature forms, although a discrete subset of intermediates appear for spliced leader RNA and tRNATyr intron splicing under La depletion conditions. 5'-end methylation in the spliced leader RNA and U1 small nuclear RNA was unaffected. The immediate cause of lethality in T. brucei was not apparent, but may represent a cumulative effect of multiple defects including processing of spliced leader RNA, tRNATyr and other unidentified RNA substrates. This study indicates that La protein binding is not essential for maturation of the spliced leader RNA, but does not rule out the presence of an alternative processing pathway that could compensate for the absence of normally-associated La protein. PMID- 15777913 TI - Prevalence, intensity and aggregation of intestinal parasites in mountain hares and their potential impact on population dynamics. AB - Mountain hare Lepus timidus populations show unstable dynamics and since hares carry a significant helminth infection and host-parasite interactions are known to be destabilising, they have been proposed as a possible causal mechanism for the observed instability. We assessed the prevalence, intensity of infection and aggregation of the helminth parasites Graphidium strigosum and Trichostrongylus retortaeformis recovered from 589 mountain hares culled from 30 Scottish sporting estates in 1999 and 2000. Graphidium strigosum showed low prevalence and intensity of infection and was highly aggregated. In contrast, T. retortaeformis showed high prevalence and intensity of infection and a low degree of aggregation. Differences in body condition of the hares were best explained by a model including sex and month of collection and interaction terms for sex-month and intensity of infection of T. retortaeformis-month. The low degree of aggregation of T. retortaeformis and the significant negative effect of intensity of infection on body condition are in accordance with the hypothesis that the host-parasite interaction is the causative destabilising mechanism for mountain hare dynamics. PMID- 15777914 TI - A dynamic population model to investigate effects of climate on geographic range and seasonality of the tick Ixodes scapularis. AB - A dynamic population model of Ixodes scapularis, the vector of a number of tick borne zoonoses in North America, was developed to simulate effects of temperature on tick survival and seasonality. Tick development rates were modelled as temperature-dependent time delays, calculated using mean monthly normal temperature data from specific meteorological stations. Temperature also influenced host-finding success in the model. Using data from stations near endemic populations of I. scapularis, the model reached repeatable, stable, cyclical equilibria with seasonal activity of different instars being very close to that observed in the field. In simulations run using data from meteorological stations in central and eastern Canada, the maximum equilibrium numbers of ticks declined the further north was the station location, and simulated populations died out at more northerly stations. Tick die-out at northern latitudes was due to a steady increase in mortality of all life stages with decreasing temperature rather than a specific threshold event in phenology of one life stage. By linear regression we investigated mean annual numbers of degree-days >0 degrees C (DD>0 degrees C) as a readily mapped index of the temperature conditions at the meteorological stations providing temperature data for the model. Maximum numbers of ticks at equilibrium were strongly associated with the mean DD>0 degrees C (r2>0.96, P<0.001), when the Province of origin of the meteorological station was accounted for (Quebec>Ontario, beta=103, P<0.001). The intercepts of the regression models provided theoretical limits for the establishment of I. scapularis in Canada. Maps of these limits suggested that the range of southeast Canada where temperature conditions are currently suitable for the tick, is much wider than the existing distribution of I. scapularis, implying that there is potential for spread. Future applications of the model in investigating climate change effects on I. scapularis are discussed. PMID- 15777915 TI - Theileria parva seroprevalence in traditionally kept cattle in southern Zambia and El Nino. AB - Sero-epidemiological surveys involving 27,526 cattle over a period of 8 years show that Theileria parva, the parasite causing East Coast fever (ECF) is found throughout southern Zambia. Higher values of T. parva sero-prevalence were observed in the plateau districts of Monze, Choma and Mazabuka than in the valley districts of Siavonga and Sinazongwe. Our results reveal a strong association between high T. parva sero-prevalence and the presence of the periodic climatic phenomenon known as the El Nino Southern Oscillation. More T. parva sero-positive samples were recorded during El Nino years (1997/98) (P<0.001) than other years in the study period. From this association, we conclude that Multiple El Nino Southern Oscillation Indices can be used to predict years with high or low ECF infection prevalence thereby contributing to the improved control of ECF in the area. PMID- 15777916 TI - Direct comparison of selected methods for genetic categorisation of Cryptosporidium parvum and Cryptosporidium hominis species. AB - A study was undertaken to compare the performance of five different molecular methods (available in four different laboratories) for the identification of Cryptosporidium parvum and Cryptosporidium hominis and the detection of genetic variation within each of these species. The same panel of oocyst DNA samples derived from faeces (n=54; coded blindly) was sent for analysis by: (i) DNA sequence analysis of a fragment of the HSP70 gene; (ii) DNA sequence analysis and the ssrRNA gene in laboratory 1; (iii) single-strand conformation polymorphism analysis of part of the ssrRNA; (iv) SSCP analysis of the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA region in laboratory 2; (v) 60 kDa glycoprotein (gp60) gene sequencing with prior species determination using PCR with restriction fragment length polymorphism analysis of the ssrRNA gene in laboratory 3; and (vi) multilocus genotyping at three microsatellite markers in laboratory 4. For detecting variation within C. parvum and C. hominis, SSCP analysis of ITS-2 was considered to have superior utility and determined 'subgenotypes' in samples containing DNA from both species. SSCP was also most cost effective in terms of time, cost and consumables. Sequence analysis of gp60 and microsatellite markers ML1, ML2 and 'gp15' provided good comparators for the SSCP of ITS-2. However, applicability of these methods to other Cryptosporidium species or genotypes and to environmental samples needs to be evaluated. This trial provided, for the first time, a direct comparison of multiple methods for the genetic characterisation of C. parvum and C. hominis samples. A protocol has been established for the international distribution of samples for the characterisation of Cryptosporidium. This can be applied in further evaluation of molecular methods by investigation of a larger number of unrelated samples to establish sensitivity, typability, reproducibility and discriminatory power based on internationally accepted methods for evaluation of microbial typing schemes. PMID- 15777917 TI - Real time PCR strategy for the identification of major lineages of Trypanosoma cruzi directly in chronically infected human tissues. AB - Two evolutionary lineages, called Trypanosoma cruzi I and II, have been identified in T. cruzi, the etiologic agent of human Chagas disease. Here, we describe a molecular strategy for direct genetic typing of these major groups of T. cruzi directly in human tissues. The protocol is based on heminested PCR amplification of the D7 region of the 24Salpha ribosomal DNA (rDNA), followed by identification of the products using denaturation curves in real time PCR. The repetitive nature of the gene, and the heminested PCR format insured the high sensitivity necessary to detect the presence of the very scarce T. cruzi DNA present in the chronically infected human tissues. There is 80% DNA sequence homology between the two 24Salpha rDNA alleles that define the T. cruzi I and II groups, sufficient to produce different thermal denaturation curves with melting temperature (TM) values of 81.7+/-0.43 and 78.2+/-0.33 degrees C (mean+/-SEM). Using this technical approach, we analysed tissue samples (esophagi, hearts and colon) from 25 different patients with the gastrointestinal or cardiac forms of Chagas disease; in all of them we found only the presence of T cruzi II. Previous epidemiological and immunological findings had already led to the idea that chronic human infections occurring in Brazil and Argentina might be primarily due to T. cruzi II strains, but all the evidence available had been indirect. Our findings provide definitive proof of this hypothesis and will also allow the establishment of which group of T. cruzi is responsible for Chagas disease in other countries. PMID- 15777918 TI - Ploidy changes associated with disruption of two adjacent genes on Leishmania major chromosome 1. AB - Leishmania major Friedlin (LmjF) is a kinetoplastid protozoan whose genomic sequence has been recently elucidated. About 60% of the identified genes do not have a known function, and many are trypanosomatid-specific. Here we characterise two adjacent genes from LmjF chromosome 1 (chr1): LmjF01.0750, which encodes a predicted protein with a serine/threonine protein kinase motif and LmjF01.0760, which encodes a product with no similarity to other known proteins. Orthologues of both genes are present in Trypanosoma cruzi, but neither occur in Trypanosoma brucei. We have mapped polyadenylation and spliced-leader acceptor sites for both genes, and show that they differ between Leishmania species. Attempts to generate null mutants of LmjF01.0750 by homologous recombination were unsuccessful and led to the apparent triploidy of the entire genome, suggesting that it is an essential gene. Interestingly, at least two copies of LmjF01.0750 are required for cell survival. Further evidence of genome plasticity in Leishmania was provided by changes in chr1 copy number that occurred during in vitro growth of wild-type LmjF promastigotes and following replacement of a single copy of LmjF01.0760. PMID- 15777919 TI - A new lineage of trypanosomes from Australian vertebrates and terrestrial bloodsucking leeches (Haemadipsidae). AB - Little is known about the trypanosomes of indigenous Australian vertebrates and their vectors. We surveyed a range of vertebrates and blood-feeding invertebrates for trypanosomes by parasitological and PCR-based methods using primers specific to the small subunit ribosomal RNA (SSU rRNA) gene of genus Trypanosoma. Trypanosome isolates were obtained in culture from two common wombats, one swamp wallaby and an Australian bird (Strepera sp.). By PCR, blood samples from three wombats, one brush-tailed wallaby, three platypuses and a frog were positive for trypanosome DNA. All the blood-sucking invertebrates screened were negative for trypanosomes both by microscopy and PCR, except for specimens of terrestrial leeches (Haemadipsidae). Of the latter, two Micobdella sp. specimens from Victoria and 18 Philaemon sp. specimens from Queensland were positive by PCR. Four Haemadipsa zeylanica specimens from Sri Lanka and three Leiobdella jawarerensis specimens from Papua New Guinea were also PCR positive for trypanosome DNA. We sequenced the SSU rRNA and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) genes in order to determine the phylogenetic positions of the new vertebrate and terrestrial leech trypanosomes. In trees based on these genes, Australian vertebrate trypanosomes fell in several distinct clades, for the most part being more closely related to trypanosomes outside Australia than to each other. Two previously undescribed wallaby trypanosomes fell in a clade with Trypanosoma theileri, the cosmopolitan bovid trypanosome, and Trypanosoma cyclops from a Malaysian primate. The terrestrial leech trypanosomes were closely related to the wallaby trypanosomes, T. cyclops and a trypanosome from an Australian frog. We suggest that haemadipsid leeches may be significant and widespread vectors of trypanosomes in Australia and Asia. PMID- 15777920 TI - Field evaluation of anthelmintic drug sensitivity using in vitro egg hatch and larval motility assays with Necator americanus recovered from human clinical isolates. AB - A field-applicable assay for testing anthelmintic sensitivity is required to monitor for anthelmintic resistance. We undertook a study to evaluate the ability of three in vitro assay systems to define drug sensitivity of clinical isolates of the human hookworm parasite Necator americanus recovered from children resident in a village in Madang Province, Papua New Guinea. The assays entailed observation of drug effects on egg hatch (EHA), larval development (LDA), and motility of infective stage larvae (LMA). The egg hatch assay proved the best method for assessing the response to benzimidazole anthelmintics, while the larval motility assay was suitable for assessing the response to ivermectin. The performance of the larval development assay was unsatisfactory on account of interference caused by contaminating bacteria. A simple protocol was developed whereby stool samples were subdivided and used for immediate egg recovery, as well as for faecal culture, in order to provide eggs and infective larvae, respectively, for use in the egg hatch assay and larval motility assay systems. While the assays proved effective in quantifying drug sensitivity in larvae of the drug-susceptible hookworms examined in this study, their ability to indicate drug resistance in larval or adult hookworms remains to be determined. PMID- 15777921 TI - Recombinant RoTat 1.2 variable surface glycoprotein as antigen for diagnosis of Trypanosoma evansi in dromedary camels. AB - The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni (insect) cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared with the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively, 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. PMID- 15777922 TI - The role of perception, language, and preference in the developmental acquisition of basic color terms. AB - When learning basic color vocabulary, young children show a selective delay in the acquisition of brown and gray relative to other basic color terms. In this study, we first establish the robustness of this finding and then investigate the extent to which perception, language, and color preference may influence color conceptualization. Experimental tasks were designed to measure different aspects of perceptual color processing (discrimination and saliency), color preference and objective counts of color term frequency in preschool-directed language (books and mothers' speech) were used to compare the acquisition of three groups of colors: primary colors, secondary colors (orange, pink, and purple) that appear at the same time as the primary colors, and secondary colors (brown and gray) that appear late. Although our results suggest that perception does not directly shape young children's color term acquisition, we found that children prefer brown and gray significantly less than basic colors and that these color terms appear significantly less often in child-directed speech, suggesting that color preference, linguistic input, and developing color cognition may be linked. PMID- 15777923 TI - Children's working memory: investigating performance limitations in complex span tasks. AB - Three experiments investigated the roles of resource-sharing and intrinsic memory demands in complex working memory span performance in 7- and 9-year-olds. In Experiment 1, the processing complexity of arithmetic operations was varied under conditions in which processing times were equivalent. Memory span did not differ as a function of processing complexity. In Experiment 2, complex memory span was assessed under three conditions designed to vary both processing and intrinsic storage demands: mental arithmetic (significant attentional demands-requires storage), odd/even judgments (significant attentional demands-no storage required), and articulatory suppression (minimal attentional demands--no storage required). The highest memory spans were found in the articulatory suppression task. Span was at an intermediate level with arithmetic processing and was lowest for processing involving odd/even judgments. This difference in memory span for processing tasks involving arithmetic processing and odd/even judgments was eliminated in Experiment 3 when the pacing requirements of the arithmetic and odd/even processing tasks were equated. The results are consistent with the view that complex memory span performance is disrupted by processing activities that divert attentional resources from storage. PMID- 15777924 TI - Grammatical sensitivity: its origins and potential contribution to early word reading skill. AB - A three-phase longitudinal study examined the origins of grammatical sensitivity and its usefulness as a predictor of early word-level reading. At about 4 years of age, children were given a range of language and cognitive tests. One year later, the children were given a further series of language and cognitive tests, this time including grammatical sensitivity, phonological sensitivity, and nonword repetition. Another year later, word-level reading achievement was assessed. Overall, grammatical sensitivity and phonological sensitivity were more firmly grounded in earlier language ability than in cognitive ability. Phonological sensitivity and nonword repetition showed reliable predictive associations with subsequent word reading skills. Grammatical sensitivity did not. PMID- 15777925 TI - The influence of provocateurs' emotion displays on the social information processing of children varying in social adjustment and age. AB - This study examined the effect of provocateurs' emotion displays on first through fourth graders' social information processing (SIP). Rating and nomination sociometric techniques were used to identify rejected-aggressive, rejected nonaggressive, average-nonaggressive, and popular-nonaggressive groups. Children viewed videotaped ambiguous provocation situations in which provocateurs' emotion displays were varied systematically. After each vignette, children's recall of story details, attribution of intent, and problem-solving responses were obtained. Half of the children were randomly assigned to a condition where they were asked directly about provocateurs' emotions, and the remaining children were not asked about provocateurs' feelings. Results revealed both developmental and social adjustment differences in SIP. In addition, provocateurs' emotion displays and asking versus not asking about those displays influenced multiple aspects of SIP. PMID- 15777926 TI - Gammadelta T-lymphocyte cytotoxic activity against Mycobacterium bovis analyzed by flow cytometry. AB - Gamma Delta (gammadelta) T lymphocytes contain the unique capability of responding to pathogens in both an innate and acquired immune response. Previously, gammadelta lymphocytes have been reported to respond to Mycobacteria tuberculosis determined by proliferation and IFN-gamma production. Unlike alpha beta (alphabeta) lymphocytes, gammadelta lymphocytes constitutively express a natural killer receptor providing gammadelta lymphocytes the capability for innate cytolytic functions. A new cytolytic assay by flow cytometry was reported capable of determining natural killer activity using K562 cells as targets without the need for radioactive materials. The objectives of this study were to first apply the flow cytometer-based assay to assess gammadelta lymphocytes natural killer activity following animal vaccination with Mycobacterium bovis Bacillus Calmette-Guerin (BCG). Secondly, to optimize the flow cytometer assay in order to detect antigen specific cytolytic activity to mycobacterium and to compare the cytolytic activity of gammadelta lymphocytes to CD-8 lymphocytes. gammadelta lymphocytes increased in NK activity (P=0.012) following animal vaccination with M. bovis BCG. Both innate (P=0.02) and acquired antigen-specific cytolytic activity (P=0.04) increased following incubation with M. bovis-infected monocytes. In conclusion, flow cytometric-based assay is a sensitive and reliable tool to determine cytolytic activity of gammadelta T-lymphocytes against mycobacterium. PMID- 15777927 TI - Detection of immune-complexed 9-O-acetylated sialoglycoconjugates in the sera of patients with pediatric acute lymphoblastic leukemia. AB - Although childhood acute lymphoblastic leukemia (ALL) is highly responsive to chemotherapy, reliable techniques are needed to determine treatment outcome. Over expression of 9-O-acetylated sialoglycoconjugates (9-OAcSGs) on lymphoblasts and concomitant anti-9-OAcSGs was found to have a diagnostic and prognostic potential. However, the presence of circulatory immune-complexed antigens remains unknown. The present study was aimed to evaluate whether immune-complexed 9 OAcSGs can be harnessed for better disease management. Immune-complexed antigens were evaluated in ALL sera (n=262) by a Dot-blot using a 9-OAcSAalpha2-6GalNAc specific lectin, Achatinin-H. Using three serum samples, the inter- and intra assay imprecision was evaluated as 11-13% and 7-11%, respectively. The recovery of spiked 9-OAcSGs was 84.2-95.4%. The central 95% reference interval for immune complexed 9-OAcSGs in normal human sera (NHS, n=144) was 2.9-3.4 mug/ml irrespective of sex and age. At disease presentation, the immune-complexed 9 OAcSGs were fivefold higher than NHS, decreased with remission induction and importantly, reappeared with clinical relapse. Sera from patients with other hematological disorders (n=86) showed negligible levels. The Dot-blot demonstrated the potential application of immune-complexed antigen as a disease specific marker and its efficacy as a sensitive and specific method that could serve as an economical yet effective index for monitoring disease status. PMID- 15777928 TI - The use of targeted microbeads for quantitative analysis of the phagocytic properties of human monocyte-derived dendritic cells. AB - The relationship between phagocytic capacity and morphology of dendritic cells (DCs) has not been investigated previously. Therefore, in order to approach this question, we have developed a novel assay, which is described here. The model of dendritic cells (DCs) used was based upon cytokine-induced differentiation of peripheral blood mononuclear cells, followed by culture on a fibronectin substratum. Under these conditions, standard current methods of quantifying phagocytosis are not applicable, as they rely upon flow cytometric analysis of fluid phase cells; and for adherent cells, quantitative efficiency of uptake is very difficult to measure. Furthermore, for both fluid phase and adherent cells, it is difficult to discriminate between internal and externally bound probe, and degradation of internalised probes can lead to artefacts. Therefore, in this study, these technical issues have been overcome by a simple and flexible assay. Phycoerythrin (PE)-conjugated antibodies are used to target microbeads to the DCs. Following an appropriate incubation period, secondary staining with fluorescein isothiocyanate (FITC)-conjugated antibody allows discrimination between internal and externally bound beads. Microscopic visualisation allows individual beads to be studied easily and thus phagocytosis quantified, whilst permitting parallel examination of morphological parameters. In particular, the relationship between bead uptake and the nature and distribution of the dendritic processes can be evaluated. PMID- 15777929 TI - Biophotonic cytotoxicity assay for high-throughput screening of cytolytic killing. AB - We have developed a highly sensitive biophotonic luciferase assay as an alternative to (51)Cr-release for assessment of cell-mediated cytotoxicity. The luciferin/ATP-dependent luminescent signal of target cells stably or transiently transfected with a firefly luciferase reporter gene (fLuc:Zeo) linearly correlates with viable target cell number. Upon incubation of fLuc:Zeo(+) target cells with CD8(+) CTLs, a rapid decrease in bioluminescence was detected that correlated with antigen-specific target cell lysis. The levels of specific lysis measured by (51)Cr-release assays correlated with the attenuation in biophotonic target cell signal, thus validating this approach as a sensitive and accurate method for the measurement of cytolysis. We show that this luminescent-based cytolytic assay (LCA) is amenable for high-throughput screening of effector cell cytolytic activity, allows for the rate of cytolysis to be measured in a single micro-plate, and permits the multiplexing of cytolytic killing with other lymphocyte functional assays such as cytokine release. Importantly, this method accurately measures the cytolytic killing of target cells that are either stably or transiently transfected with a fLuc reporter gene, and thus is ideal for monitoring cytolysis of both primary autologous and immortalized target cell lines. The versatility of the non-radioactive, high-throughput, biophotonic cytolytic assay should make this method an attractive alternative to chromium release for quantifying effector cell cytolytic activity. PMID- 15777930 TI - Activation-dependent phases of T cells distinguished by use of optical tweezers and near infrared Raman spectroscopy. AB - Near-infrared Raman spectroscopy may provide a highly sensitive, noninvasive means to identify activation status of leukocytes. The purpose of the current study was to establish Raman spectroscopic characteristics of T cell activation. Activation of the RsL.11 T cell clone in vitro with Con A resulted in specific decrements in band intensities at 785, 1048, 1093, and 1376 cm(-1) but did not alter a majority of other band intensities including those at 1004 cm(-1) (phenylalanine) and 1660 cm(-1) (amide bonds). Activation-dependent decrements in these band intensities occurred subsequent to IL-2 production and correlated closely with T cell blastogenesis. Activation-dependent decrements in these band intensities were not strictly a function of cell size because the same observations were noted in size-controlled comparisons of resting and activated T cells. Like the RsL.11 clone, freshly isolated thymocytes that were activated by Con A or IL-2 showed decrements in particular emissions. These findings indicate that near-infrared Raman spectroscopy can be used as a noninvasive technique to reveal the activation status of single living T cells. PMID- 15777931 TI - Flow cytometry identification and characterization of mononuclear cell subsets in the neotropical primate Saimiri sciureus (squirrel monkey). AB - BACKGROUND: The neotropical primate squirrel monkey is used in many areas of biomedical research including neuroendocrinology, immunology and infectious diseases. However, research has been hampered by the lack of immunological tools for this primate. METHODS: A series of 67 commercially available monoclonal antibodies to human CD antigens or cytokines were tested on Saimiri mononuclear cells and the specificity was assessed by double staining using flow cytometry. RESULTS: Monoclonal antibodies defining the main mononuclear cells subsets (monocytes, B, T, including CD4 and CD8 T cells) as well as activation markers have been identified. The conditions to specifically identify the various cell subsets using two color flow cytometry and establish their relative proportions have been set-up. We also have established normal values of the main circulating mononuclear cell subsets for adult Saimiri sciureus monkeys from the breeding unit of Institut Pasteur in French Guiana. The distribution between spleen, blood and lymph nodes has been compared. CONCLUSIONS: These tools allow documenting the phenotype of most Saimiri mononuclear cell subsets and assessing their activation level. This opens new perspectives for vaccinology and immunopathology research in this experimental non-human primate host, in particular for malaria research. PMID- 15777932 TI - Ribosome display of antibodies: expression, specificity and recovery in a eukaryotic system. AB - In ribosome display, proteins are linked to their encoding genetic material as protein-ribosome-mRNA complexes. The technology has been applied to the isolation of antibodies and other proteins from large PCR-derived libraries. Here we demonstrate the specificity of eukaryotic ribosome complexes and investigate recovery and display procedures using a single chain version of the anti progesterone monoclonal antibody DB3. Complexes are formed by deletion of the 3' stop codon in a coupled rabbit reticulocyte system. Using inhibition with different steroid probes, we show that the fine specificity of the combining site expressed as a nascent protein is closely similar to the native monoclonal, indicating correct folding and function while bound to the ribosome. We have demonstrated that the 3' end of the mRNA is blocked by the stalled ribosome and unavailable to primers. Moreover, we show that an in situ RT-PCR recovery procedure, carried out on intact complexes, is more efficient than ribosome disruption and isolation of mRNA followed by RT-PCR. We also explore the Mg(2+) and DTT concentrations and time required for efficient production of complexes. Our findings confirm the effectiveness of the eukaryotic ribosome display system and define conditions for efficient selection of single chain antibodies. PMID- 15777933 TI - Single-step purification of human C4b-binding protein (C4BP) by affinity chromatography on a peptide derived from a streptococcal surface protein. AB - Many Gram-positive bacteria express surface proteins that bind human plasma proteins. These bacterial proteins, and derivatives of them, are of interest for analysis of bacterial pathogenesis and as immunochemical tools. Well characterized examples include the IgG-binding reagents staphylococcal protein A and streptococcal protein G, and the recently described streptococcal IgA-binding peptide Sap. Here, we show that a peptide derived from the streptococcal M22 protein can be used for single-step affinity purification of the human complement regulator C4b-binding protein (C4BP). Binding of C4BP was strongly enhanced by dimerization of the peptide via a C-terminal cysteine residue not present in the intact M22 protein. The purified C4BP had the expected binding characteristics, and acted as a cofactor for factor I in the degradation of C4b. Passage of serum through a peptide column under non-saturating conditions resulted in binding of >99.5% of serum C4BP, implying that such a column can be used to deplete serum of C4BP. These data indicate that the C4BP-binding peptide is a versatile tool that can be used for simple and rapid purification of biologically active human C4BP or for removal of C4BP from serum. PMID- 15777934 TI - Evaluating ELISPOT summary measures with criteria for obtaining reliable estimates. AB - The ELISPOT assay is a commonly used technique for quantifying the occurrence of T lymphocyte cells secreting a cytokine after stimulation with an antigen or peptide. The assay endpoint, the number of spot-forming cells (SFC) at a specific concentration of effector cells, is typically estimated using either a simple arithmetic mean or the predicted value from a linear regression model. We compare statistical modeling approaches for summarizing these assays using data from the Pediatric AIDS Clinical Trial Group (PACTG) study 299. A simulation study was conducted to compare methods under controlled conditions. Assuming the optimal effector cell concentration is known, we demonstrate that the simple mean is appropriate if assays are conducted at the same concentration for all samples. Normalizing simple means to a summary concentration using results from a range of concentrations is not valid. A random effects or mixed model is superior to the simple mean when a large within-assay (subject) variance relative to between subject variance exists. When assays are conducted over a range of effector cell concentrations for each individual, the theoretical linearity assumption of the regression model is often violated and can result in biased estimates. In this case, nonlinear models provide more accurate estimation. Collecting data over a range of concentrations allows reassessment of the optimal cell concentration after the data are generated. PMID- 15777935 TI - Sheep red blood cells armed with anti-CD20 single-chain variable fragments (scFvs) fused to a glycosylphosphatidylinositol (GPI) anchor: a strategy to target CD20-positive tumor cells. AB - Single-chain variable fragment antibodies (scFv) retain antigen specificity and offer advantages over intact antibodies as therapeutic agents. We cloned the cDNA of the V(H) and V(kappa) regions from a mouse hybridoma (HB-9645) directed against human CD20. In addition to the basic scFv construct (V(kappa)-L-V(H)), we genetically engineered a secretory signal, six histidine residues, and a 'Flu' tag to facilitate secretion, purification, and detection. A glycosyl phosphatidylinositol (GPI) modification signal was added at the C terminus. The GPI-tagged and the non-tagged scFvs were expressed in high yields on the surface of stably transfected insect cells. The CD20-binding properties of purified non GPI tagged scFv were examined using flow cytometry and immunocytochemistry. The non-GPI-tagged scFv selectively recognizes CD20-positive cells in a concentration dependent manner. Double-flow cytometry analysis using fresh peripheral blood lymphocytes and WSU-FSCCL cells revealed that our scFv resolves the B-cell population better than the intact antibody. The GPI-tagged scFv was loaded onto the surface of sheep erythrocytes to form rosettes with CD20-positive cells. The genetically engineered anti-CD20 scFv and GPI-tagged derivative have binding specificity for the CD20 antigen. The scFvs described here has potential uses as an in vivo tumor-imaging agent and as a carrier vehicle for targeted delivery of cytocidal agents to CD20-positive cancer cells. PMID- 15777936 TI - Enhanced sensitivity of surface plasmon resonance (SPR) immunoassays using a peroxidase-catalyzed precipitation reaction and its application to a protein microarray. AB - We describe a method to improve the sensitivity of surface plasmon resonance (SPR) immunoassays using a horseradish peroxidase (HRP)-catalyzed precipitation reaction. The precipitation reaction catalyzed by HRP bound to the SPR biosensor surface via a sandwich immunoassay induced a shift in the SPR angle. Human interferon (IFN)-gamma at concentrations ranging from 0.01 to 100 ng/ml was detectable by this method. We also show that this biocatalytic signal amplification method can be applied to SPR imaging (SPRI), in an immunoassay of multiple proteins on a protein microarray format. PMID- 15777937 TI - Development of an assay for the quantification of type I collagen synthesis in the guinea pig. AB - There is a need for a reliable assay for the quantification of collagen type I synthesis in the guinea pig, an important model for many connective tissue diseases. Procollagen type I C-terminal propeptide (PICP) is the established marker of type I collagen synthesis but, to date, no assay has been developed to measure PICP in guinea pig tissue extracts. A monoclonal antibody, known to cross react with intact guinea pig procollagen type I (anti-PICP), was tested for its ability to bind soluble guinea pig PICP in crude skin extracts using a biosensor. Anti-PICP was immobilised to the surface of a sensor chip and antibody-antigen binding was detected using the phenomenon of surface plasmon resonance (SPR). The binding component in the SPR-immunoassay was identified as PICP by purification and N-terminal sequencing. Guinea pig PICP was purified from skin by gel filtration, ion exchange chromatography and lectin affinity chromatography. Purified PICP was then biotinylated and used with anti-PICP to develop a competition ELISA that was able to selectively and sensitively measure PICP in extracts of guinea pig connective tissue. PMID- 15777938 TI - Analysis of the binding of monoclonal and polyclonal antibodies to the glycoproteins of antigenic variants of human respiratory syncytial virus by surface plasmon resonance. AB - The surface glycoproteins of human respiratory syncytial virus (hRSV), F and G, are the major protective antigens of the virus. Both are antigenically variable, although to different degrees, but the role of antigenic variation in the pathogenesis of hRSV disease has not been fully evaluated. Assessment of immunity to different virus strains is difficult with conventional antibody assays where differing properties of the virus antigens, other than antigenicity, may influence the outcome of the assay. Here, we have developed BIAcore surface plasmon resonance based assays for antibodies to the glycoproteins of hRSV which allow valid comparison of antibody titres against multiple hRSV strains. Glycoproteins from a number of lineages of hRSV sub-group A were captured from lysates of infected cells onto the dextran coated surface of a BIAcore sensor chip via primary monoclonal antibodies (MAbs) to conserved epitopes. For the G glycoprotein, primary MAbs were conjugated directly to the dextran of the sensor chip via free amide groups. For the F glycoprotein, direct conjugation was found to inactivate the MAb and primary MAb was immobilised on the chip via rabbit anti mouse Fc antibody fragments in an indirect system. Using monoclonal antibodies as secondary MAbs, the glycoproteins in both systems were shown to exhibit a sub-set of conserved and variable epitopes, with some epitopes of both sorts being unavailable, presumably blocked by the primary antibody. Polyclonal anti-hRSV sera raised against viruses of different genotype bound equally to both F and G glycoproteins from homologous and heterologous viruses suggesting that mice immunised systemically with lysates of cells infected with recent isolates of virus do not respond well to genotype specific epitopes. PMID- 15777939 TI - Identification of HLA-DRB1-bound self-peptides following measles virus infection. AB - We developed a B-lymphocyte cell line derived from a measles seropositive individual who was homozygous for the HLA-DRB1*0301 allele. Peptides associated with the HLA-DRB1*0301 protein were purified from this lymphoblastoid cell line after infection with the Attenuvax measles vaccine virus (Merck Research Laboratories, West Point, PA) and with "sham" infection. More than 40 peptide sequences were obtained by nano-scale reversed phase-high performance liquid chromatography coupled to tandem mass spectrometry (nano-LC/MS/MS). These peptides originated from 21 different source proteins--the majority from membrane bound proteins. Most of the peptides (>73%) bound to HLA-DRB1*0301 appeared to be in lower abundance on measles-infected cells compared to the "sham-infected" cells. However, 26% of the identified peptides seem to have increased expression after measles infection. Measles vaccine virus infection did not change the level of HLA-DR expression. We demonstrate the power of nano-LC/MS/MS in the rapid determination of changes in the spectrum and expression of HLA-DRB1*0301-bound peptides after infection with measles virus. This provides further knowledge of the changes in peptide expression after viral infection and provides a powerful tool for identifying HLA-presented host and viral epitopes in the context of class II HLA molecules. PMID- 15777940 TI - Flow-cytometric screening of platelet antibodies with previously frozen cells. AB - In this study the flow-cytometric crossmatch results were compared between fresh cells and cells processed by various cryopreservation and storage methods. Platelets from healthy donors were incubated with 12 sera containing platelet reactive antibodies as well as with 62 control sera from blood donors. Direct comparisons were made between fresh platelets and platelets after freezing at -28 degrees C, -40 degrees C and -80 degrees C and in liquid nitrogen, using 6% DMSO as cryoprotectant. In addition, the effects of using controlled-rate freezing were evaluated. Finally we evaluated the application of the cryoprotectant ThromboSol. The best results were obtained after cryopreservation of the platelets with ThromboSol at -80 degrees C with controlled cooling rates. Using ThromboSol cryopreserved platelets, the sensitivity for the detection of incompatible platelets was 100% and the specificity was 97.1%, using the previous results obtained with flow-cytometry, MAIPA and LCT as a reference. CONCLUSION: Platelets can be frozen using ThromboSol as the cryoprotectant, with controlled rate freezing and storage at -80 degrees C for the screening of platelet antibodies and for flow-cytometric crossmatch procedures. This system yields a reproducible and logistically simple method for platelet crossmatching that yields results superior to fresh cells and can be easily incorporated into standard clinical laboratory practices. PMID- 15777941 TI - Granulocyte contamination dramatically inhibits spot formation in AIDS virus specific ELISpot assays: analysis and strategies to ameliorate. AB - The interferon-gamma (IFNgamma) ELISpot assay has become the most critical tool for HIV vaccine evaluation. External factors affecting ELISpot results must be minimized for the data to be reliably used in vaccine research and development processes. In pre-clinical pigtail macaque studies analyzing HIV/SIV vaccine studies, we detected a strong correlation between levels of granulocytes contaminating PBMC preparations and reduction in the quality and quantity of spots in the IFNgamma ELISpot assay. Acute SHIV infection of macaques worsened granulocyte contamination of PBMC fractions and made the assay much less reliable in detecting SIV-specific T cell immunity compared to intracellular cytokine staining (ICS). This problem could be ameliorated by using an F(ab)(2) form of the MD-1 IFNgamma capture antibody, presumably reflecting that activation of granulocytes in the well by the Fc portion of the standard capture antibody disrupts spot formation. Improving the standard ELISpot assay by using an F(ab)(2) capture antibody should make it more reliable for use in critical vaccine development studies. PMID- 15777942 TI - Molecular characterization of the circulating anti-HIV-1 gp120-specific B cell repertoire using antibody phage display libraries generated from pre-selected HIV 1 gp120 binding PBLs. AB - Human bone marrow is a major repository for maturated antibody-secreting plasma cells, which produce the majority of the antibodies found in serum, making it an attractive source for generating human immune antibody libraries. Unfortunately, bone marrow is not always readily available and, although human immune libraries can be generated from circulating B cells, the low frequency of antigen-specific B cells in the circulation yield few monoclonal antibodies of interest. We used a pre-selection strategy to enrich for antigen-specific B cells prior to library generation, and applied this approach to evaluate, at a molecular level, the nature of the human anti-HIV-1 gp120 repertoire encoded by circulating B cells. IgG antibody phage display libraries were generated from HIV-1 seropositive individuals using either affinity-selected anti-gp120 IgG-bearing circulating B cells, predominantly exhibiting memory/activated B cell phenotype, or unselected PBMCs. These libraries were selected against HIV-1 gp120, resulting in isolation of a panel of gp120-specific antibodies. Whereas only 2 gp120-specific antibodies were retrieved from the non-pre-selected HIV-1 library, 9 gp120-specific antibodies were retrieved from the 10-fold smaller library generated from the pre selected B cells, demonstrating the feasibility of the approach. The anti-gp120 antibodies derived from the circulating B cells of HIV-1 donors generally resembles those from bone marrow plasma cells with respect to epitope specificity, affinity and neutralization ability. They exhibit high affinity for gp120, are directed against a variety of epitopes, but rarely exhibit the ability to neutralize HIV-1. PMID- 15777943 TI - Analysis of the specificity and thermodynamics of the interaction between low affinity antibodies and carbohydrate antigens using fluorescence spectroscopy. AB - The purpose of this work has been to examine whether fluorescence spectroscopy can be used to investigate weak or transient binding between monoclonal antibodies and carbohydrate antigens. In earlier studies we have demonstrated that the three monoclonal antibodies 39.4 (IgG2b), 39.5 (IgG2b) and 61.1(IgG3) bind weakly to the glycosidic alpha(1-4) bond present in e.g. maltose and panose. In this study these antibodies showed an enhancement in the fluorescence intensity of tryptophan upon binding in solution to these two carbohydrate antigens. Using a structural analog to maltose, cellobiose, no fluorescence intensity change was induced. Dissociation constants for these antibodies at different temperatures (5-40 degrees C) were obtained in the range of 0.003-0.2 mM and they were in accordance with earlier data from studies on affinity chromatography and surface plasmon resonance. Almost a doubling of the dissociation constants was observed for every 10 degrees C increase in temperature, giving an exothermal reaction with standard enthalpy change of -51 kJ/mol, for the association between antibody and carbohydrate antigen. It was seen that the extra glycosyl ring in panose increased the affinity more than eight times for the monoclonal antibody 39.5. A standard entropy increase of 21%, probably due to hydrophobic effects, is introduced by the extra glycosyl ring, while the enthalpy stays unaffected. This direct fluorescence approach to measure the binding and thermodynamics of an interacting antigen-antibody pair is simple and accurate since measurements are performed in solution and no immobilization or fluorophore labeling of the components is required. Introduction of fluorescence techniques will be a useful complement to current procedures to measure interaction of antibody with antigen and in particular they will offer solutions to detect transiently binding antigens. PMID- 15777944 TI - Affinity maturation of a V(H)H by mutational hotspot randomization. AB - V(H)Hs from naive libraries have dissociation constants (K(D)s) in the low micromolar range and thus, for most antibody applications, their intrinsic affinities need to be improved significantly. Non-targeted in vitro affinity maturation approaches based on indiscriminate randomization of complementarity determining region (CDR) residues or random mutagenesis of conventional antibody variable domains have been shown to improve the affinity of recombinant antibodies by 450- to over 6000-fold. A different, targeted approach based on selective randomization of CDR codons containing AGY/RGYW nucleotide mutational hotspots i.e., "hotspot codons", also promises to be very efficient for improving antibody affinities. Here we employed the latter approach for improving the affinity of PTH22, a parathyroid hormone (PTH)-derived peptide-specific V(H)H that was isolated from a naive llama phage display library. A PTH22 mutant ribosome display library was constructed by randomizing nine CDR2 and CDR3 hotspot codons. The affinity improvement of the lead binder was 30-fold, which seems somewhat low in view of the large number of randomized hotspot codons. Nucleotide sequence analyses of PTH22 and 23 naive V(H)Hs suggested that many AGY/RGYW mutational hotspots are not affinity mutational hotspots but play a role in V(H)H solubility, structure, and deletion/insertion events. Our results indicate that the mutagenesis approach described here is beneficial in terms of yielding moderate increases in affinity while fine-tuning physical properties of an antibody. PMID- 15777945 TI - Use of biotinylated 17beta-estradiol in enzyme-immunoassay development: spacer length and chemical structure of the bridge are the main determinants in simultaneous streptavidin-antibody binding. AB - 17beta-estradiol (E2) concentrations are in the low pg/ml range in plasma. To develop a sensitive enzyme immunoassay (EIA) for E2-determination a highly specific antibody raised against a 6-carboxymethyl (CMO)-E2-bovine serum albumine conjugate was used. Based on 6-CMO-E2 and 6-amino-E2, four biotinylated tracers with two different spacer lengths between E2 and biotin were synthesized using biotinylation reagents in one step reactions. All amino-based tracers were unsuitable for assay development because the antibody binding was too weak compared to the analyte E2. For 6-CMO-based tracers the simultaneous binding of the tracer to the antibody and streptavidin seems to be the determining step in the procedure depending on incubation temperature and spacer lengths. While a short spacer of 9 carbon atoms was susceptible to room temperature, a longer spacer of 16 carbon atoms showed nearly the same results for incubation at 4 degrees C or at room temperature. The absolute detection limit of this system was 0.63 pg/well. For sample clean-up, porcine plasma was solvent-extracted and depending on the initial plasma volume further purified by solvent partition. Determination of reproducibility resulted in intraassay coefficients of variation of 13% and 5.3% for samples with E2-levels of 15 pg/ml and 236 pg/ml, respectively. Measurement of E2-spiked blood plasma revealed recoveries of 83% up to 100% for E2 concentrations between 50 pg/ml and 1000 pg/ml. Only for the lowest concentration (20 pg/ml) a recovery of 58% was observed. Correlation of the EIA with an established radio immunoassay resulted in r=0.991 using the same antibody. PMID- 15777946 TI - An in vitro evaluation of the potential suitability of peripheral blood CD14(+) and bone marrow CD34(+)-derived dendritic cells for a tolerance inducing regimen in the primate. AB - Dendritic cells (DC) represent a tool not only for immune activation, but also potentially for tolerance induction in transplantation. This latter approach is yet to be explored in a pre-clinical primate model. Since no information concerning baboon DC has been available, we characterised the DC of this species derived in vitro from bone marrow (CD34(+)) and peripheral blood (CD14(+)) precursors to determine which would be the most suitable for a tolerance inducing strategy. Baboon DC were differentiated in vitro using protocols similar to those used in humans and their maturation status was assessed and compared according to their phenotype and function. Based on both phenotypic and functional criteria, the CD14-derived baboon DC appeared to be less mature DC, necessitating an additional stimulus in order to become fully mature. The CD34-derived DC on the other hand appeared more mature in nature, without necessarily requiring exposure to overt maturation signals. We suggest therefore that, in the baboon, peripheral blood CD14-derived DC may be more suitable for protocols where tolerance induction is the goal. We now aim to perform further in vitro investigations into the potential tolerance inducing effects of CD14-derived DC alone or in association with other strategies that would be applicable in vivo. PMID- 15777947 TI - Snake egg immunoglobulins: biochemical characteristics and adjusted isolation procedure. AB - Transmission of specific immunoglobulins from mothers to their offspring via the egg is a common phenomenon in egg-laying vertebrates but the occurrence of this phenomenon in reptiles, especially in colubrid snakes, has not been proven until recently. Thus, the essential biochemical characteristics of antibodies deposited in eggs of Elaphe guttata (Colubridae, Serpentes) were studied after isolation of the antibody by precipitation and purification by affinity chromatography and gel filtration, with verification by isoelectric focusing and immunoprecipitation. The immunoglobulin deposited in the eggs of colubrid snakes is a singular, non truncated IgY antibody in a concentration roughly equal to that in the snake's serum. An efficient method to isolate antibody from snake eggs was developed, based on the PEG precipitation technique of avian yolk immunoglobulins; an unsophisticated protocol for the isolation procedure appropriate for reptile eggs is provided. PMID- 15777948 TI - Phosphorothyoate oligodeoxynucleotides block nonspecific binding of Cy5 conjugates to monocytes. AB - Cy5-based monoclonal antibody conjugates, including those containing PE-Cy5 and Cy5 alone, are used widely for multi-color flow cytometry in both research and clinical applications. One problem with these conjugates is they bind to monocytes and macrophages irrespective of antibody specificity. This effect, which is thought to be due to binding of the Cy5 itself to the human high affinity receptor for IgG (FcgammaRI, CD64), can complicate interpretion of flow cytometric results. In the present study we demonstrate that phosphorothioate oligodeoxynucleotides (PS-ODN) suppress this nonspecific binding in a manner that is independent of PS-ODN sequence. Binding of FITC-labeled PS-ODN to monocytes was blocked by CD64-specific monoclonal antibodies, suggesting CD64 is a PS-ODN binding protein. We conclude PS-ODN can be used as an effective, simple and low priced reagent to prevent nonspecific binding of Cy5-based immunoconjugates to monocytes. This effect appears to be mediated by PS-ODN binding to CD64 thereby blocking the binding of Cy5 to this receptor. PMID- 15777949 TI - Identifying subpopulations of thymic epithelial cells by flow cytometry using a new specific thymic epithelial marker, Ly110. AB - We generated monoclonal antibodies reacting to a mouse thymic epithelial cell specific membrane protein, Thymic Stromal Co-transporter (TSCOT)/Ly110. These antibodies showed specificity to the peptide sequences derived from TSCOT/Ly110 determined by specific peptide inhibition in flow cytometric analyses with cells expressing the protein on the surface. TSCOT/Ly110 expressing subpopulation can be identified among the CDR1(+) or 6C3(+) cortical epithelial cells. Furthermore, CDR1 positive cortical thymic epithelial cells can be separated into further distinguishable populations; CDR1(+)6C3(+)Ly110(+), CDR1(+)6C3(-)/(low)Ly110(+), CDR1(+)Ly110(-). Some of TSCOT/Ly110 expressing cells negative for both CDR1 and 6C3 markers were found at the earlier stages of development, while most of the cells are positive for both at 1-week-old stage. After then, downregulation in 6C3 and/or CDR1 expression was noticed until 16 weeks of age. These results suggest that TSCOT/Ly110 is a new marker for the subpopulation of CDR1(+) or 6C3(+) epithelial cells in the neonatal and adult thymus and is useful for the studies on the epithelial cell differentiation process. PMID- 15777950 TI - Sphingomyelinase D from venoms of Loxosceles spiders: evolutionary insights from cDNA sequences and gene structure. AB - Loxosceles spider venoms cause dermonecrosis in mammalian tissues. The toxin sphingomyelinase D (SMaseD) is a sufficient causative agent in lesion formation and is only known in these spiders and a few pathogenic bacteria. Similarities between spider and bacterial SMaseD in molecular weights, pIs and N-terminal amino acid sequence suggest an evolutionary relationship between these molecules. We report three cDNA sequences from venom-expressed mRNAs, analyses of amino acid sequences, and partial characterization of gene structure of SMaseD homologs from Loxosceles arizonica with the goal of better understanding the evolution of this toxin. Sequence analyses indicate SMaseD is a single domain protein and a divergent member of the ubitiquous, broadly conserved glycerophosphoryl diester phosphodiesterase family (GDPD). Bacterial SMaseDs are not identifiable as homologs of spider SMaseD or GDPD family members. Amino acid sequence similarities do not afford clear distinction between independent origin of toxic SMaseD activity in spiders and bacteria and origin in one lineage by ancient horizontal transfer from the other. The SMaseD genes span at least 6500bp and contain at least 5 introns. Together, these data indicate L. arizonica SMaseD has been evolving within a eukaryotic genome for a long time ruling out origin by recent transfer from bacteria. PMID- 15777951 TI - How does agkicetin-C bind on platelet glycoprotein Ibalpha and achieve its platelet effects? AB - The platelet glycoprotein (GP) Ib-IX-V receptor complex has a central role in primary haemostasis and possesses binding sites for the plasmatic adhesive protein von Willebrand Factor (VWF) and thrombin. Several snake venom components have been identified in recent years that target this receptor complex and modulate its functionality. Among them, agkicetin-C is from Deinagkistrodon acutus and proved to be a potent antagonist of GPIb-IX-V. We further studied the structure-activity relationships of agkicetin-C in order to reveal the molecular mechanisms of its antagonistic effect. Agkicetin-C concentration-dependently inhibited botrocetin-, ristocetin- and low dose thrombin- (0.32-0.4nM) induced platelet aggregation. Moreover, it abolished platelet adhesion to collagen under high shear conditions (2600/s), while having only minor effects at low shear rate (650/s), which suggested it targets mainly GPIbalpha instead of other platelet glycoproteins. The interaction site of agkicetin-C was further refined: it recognizes a linear sequence in a recombinant GPIbalpha (AA1-289) fragment and inhibited completely the ristocetin-induced VWF binding to this fragment. Using cross-blocking studies with epitope-mapped anti-GPIbalpha monoclonal antibodies, the binding region of agkicetin-C was refined to the AA201-282 region. In conclusiong the C-type lectin agkicetin-C is a potent GPIb-IX-V antagonist, inhibiting both VWF and thrombin interaction through binding to the AA201-282 region in GPIbalpha. Another thing of interest is that, although agkicetin-C did not agglutinate platelets in all conditions tested in vitro, it caused a severe thrombocytopenia in rats, suggesting a different mechanism than with flavocetin-A or echicetin. PMID- 15777952 TI - Primary structure of brevilysin L4, an enzymatically active fragment of a disintegrin precursor from Gloydius halys brevicaudus venom. AB - Brevilysin L4 (L4) is a non-hemorrhagic P-I class metalloprotease (MP) isolated from Gloydius halys brevicaudus venom. Its complete amino acid sequence has been determined. L4 is a single-chain polypeptide and highly homologous to those of other snake venom MPs. A zinc-binding motif, HExxHxxGxxH, is located at residues 142-152. A characteristic feature of L4 is the presence of a spacer sequence (LRTDTVS) at the C-terminal that links metalloprotease and disintegrin domains and is usually removed by post-translational proteolysis, suggesting that L4 is expressed together with a spacer region and a disintegrin domain at the C terminal. The nucleotide sequence of a cDNA clone encoding L4 has revealed that L4 is a disintegrin precursor and produced as a P-II class MP. The disintegrin coded after L4 sequence was brevicaudin 1, a disintegrin previously isolated from the same venom. P-II class MPs have been suspected to undergo autoproteolysis to release disintegrins. Although being P-I class MP, L4 itself autocatalytically degrades with a half-life of 30min at pH 8.5 and 37 degrees C in the absence of Ca(2+). Sequence analysis of several fragment peptides produced during the autolysis of L4 indicated that more than 40 peptide bonds were split, and the cleavages of Ser(60)-Asn(61), Thr(99)-Ala(100), and Phe(103)-Asp(104) bonds may trigger the autoproteolysis. Addition of Ca(2+) completely suppressed the cleavage of these particular bonds, resulting in a marked prevention of autoproteolysis. Thus, L4 provides a good model for the investigation of autolysis of some MPs. PMID- 15777953 TI - Studies on the cardenolide sequestration in African milkweed butterflies (Danaidae). AB - Butterflies of the Danaidae family are considered to be toxic or distasteful due to the presence of cardiac glycosides sequestered from their larval food plants. Alcoholic extracts of specimens of Danaus chrysippus aegyptius and Amauris ochlea ochlea from southern Africa (Namibia, S.-Africa, Mozambique) were analyzed by thin-layer chromatography for these cardenolides. But only 4 of 75 specimens of D. chrysippus aegyptius contained trace amounts, all others including 13 specimens of A. ochlea ochlea were negative. Genetic analysis of the ouabain binding site of the Na(+), K(+)-ATPase revealed that both species do not present an amino acid replacement at the position 122, which otherwise makes the enzyme insensitive to cardenolides suggesting that other strategies of toxin tolerance must have been developed. PMID- 15777954 TI - Pharmacological investigation of the nociceptive response and edema induced by venom of the scorpion Tityus serrulatus. AB - In this study we characterized the nociceptive response and edema induced by the venom of the scorpion Tityus serrulatus in rats and mice and carried out a preliminary pharmacological investigation of the mechanisms involved in these responses. Intraplantar injection of the venom (1 or 10mug) induced edema and a marked ipsilateral nociceptive response, characterized by thermal and mechanical allodynia and paw licking behaviour. The nociceptive response was inhibited by previous intraperitoneal administration of indomethacin (4mg/kg), dipyrone (200mg/kg), cyproheptadine (10mg/kg) or morphine (5 or 10mg/kg), but not by dexamethasone (1 or 4mg/kg) or promethazine (1 or 5mg/kg). The edema was inhibited by previous treatment with promethazine (5 or 10mg/kg) or cyproheptadine (5 or 10mg/kg), but not by indomethacin (2 or 4mg/kg), dexamethasone (1 or 4mg/kg) or cromolyn (40 or 80mg/kg). Some bioactive amines, including histamine and 5-hydroxytryptamine, were found in the venom in low concentrations. In conclusion, the nociceptive response and edema induced by the venom of T. serrulatus may result from the action of multiple mediators including eicosanoids, histamine and 5-hydroxytryptamine. These results may lead to a better understanding of the host response to potent animal toxins and also give insights into a more rational pharmacological approach to alleviate the intense pain associated with the scorpion envenomation. PMID- 15777955 TI - Further isolation and characterization of grammistins from the skin secretion of the soapfish Grammistes sexlineatus. AB - Soapfishes contain peptide toxins (grammistins) in the skin secretion. Two grammistins (Gs 1 and Gs 2) and six grammistins (Pp 1, Pp 2a, Pp 2b, Pp 3, Pp 4a and Pp 4b) have already been isolated from Grammistes sexlineatus and Pogonoperca punctata, respectively. In this study, five grammistins (Gs A-E), together with grammistins Gs 1 and Gs 2, were further isolated from G. sexlineatus by gel filtration and reverse-phase HPLC. Sequence analyses revealed that grammistins Gs A (28 residues) and Gs C (26 residues) are analogous to grammistin Pp 3 and grammistin Gs B (12 residues) to grammistin Pp 1, while grammistins Gs D (13 residues) and Gs E (13 residues) are identical with grammistins Pp 1 and Pp 2b, respectively. Grammistins Gs A-C exhibited antibacterial activity with a broad spectrum against nine species of bacteria in common with the other grammistins but had no hemolytic activity differing from the other grammistins. Grammistins Gs A-E, Gs 1 and Gs 2 could release carboxyfluorescein entrapped within liposomes made of either phosphatidylcholine or phosphatidylglycerol/phosphatidylcholine (3:1), demonstrating their membrane-lytic activity. However, no clear relationship between the membrane-lytic activity and the biological activity of grammistins was recognized. PMID- 15777956 TI - Variability of alkaloids in the skin secretion of the European fire salamander (Salamandra salamadra terrestris). AB - The two major alkaloids, samandarine and samandarone, were identified in the skin secretion of individual specimens from two populations of the European fire salamander (Salamandra salamandra terrestris) by gas chromatography/mass spectrometry. High intraspecific variability in the ratio of both alkaloids was observed, but also in individual specimens over a period of 4 months suggesting separate metabolic pathways of the compounds. Alkaloid synthesis appears to take place also in liver, testes and ovaries, whereas the larvae of the salamanders are entirely free of alkaloids. PMID- 15777957 TI - Ultrasensitive detection of domoic acid in mouse blood by competitive ELISA using blood collection cards. AB - Domoic acid (DA), an analog of the excitatory amino acid glutamate, is produced by the diatom genus Pseudo-nitzschia and acts as a neurotoxin in humans. During diatom blooms, DA can contaminate shellfish, as well as other filter feeding organisms, and can be transferred by ingestion to higher trophic levels, including marine mammals and humans. The prevalence of this algal toxin and its effects on protected species makes measurement of domoic acid in living animals a necessary biomonitoring tool for the near future. Blood collection cards have already been used for the sampling, extraction and detection of brevetoxin in blood from exposed laboratory animals and, more recently, marine mammals. However, a difficulty unique to measuring DA in blood is the rapid rate (>95% in 2h) at which it is cleared from blood. To meet this challenge, a direct competitive ELISA (cELISA), a method of detection with extremely high sensitivity and specificity, was used to analyze the blood of DA-exposed mice after extraction from the blood collection cards. More than 99% of DA was cleared from blood within 4h post dosage; however, domoic acid was still quantifiable (>0.7ngml(-1)) at 4h from blood spot extracts and still detectable at 24h when compared to control blood spots. By using this highly sensitive assay in conjunction with the use of blood spot cards for easy blood sample extraction, this method could be a very effective means of biomonitoring domoic acid in marine mammals in the field, as well as human populations. PMID- 15777958 TI - Tissue distributions and seasonal dynamics of the hepatotoxic microcystins-LR and -RR in two freshwater shrimps, Palaemon modestus and Macrobrachium nipponensis, from a large shallow, eutrophic lake of the subtropical China. AB - So far no information is available on microcystin (MC) contents in shrimps, prawns or crayfish from natural freshwaters. Tissue distributions and seasonal dynamics of the hepatotoxic MC-LR and -RR in two freshwater shrimps, Palaemon modestus and Macrobrachium nipponensis were studied monthly (during June November, 2003) in a Chinese lake containing toxic cyanobacterial blooms. The shrimps P. modestus and M. nipponensis accumulated high MCs not only in the hepatopancreas (mean 4.29 and 0.53 microg g(-1) DW, respectively) but also in the gonad (mean 1.17 and 0.48 microg g(-1) DW, respectively), and the crayfish Procambarus clarkii accumulated as much as 0.93 microg g(-1) DW in the gonad. This indicates that gonads of these invertebrates are the second important target organ of MCs. P. modestus apparently accumulated more MCs in their organs than M. nipponensis, which might be a reflection of their difference in trophic niche. Eggs of the shrimps accumulated 8.4% (M. nipponensis, 0.27 microg g(-1) DW) and 29.0% (P. modestus, 2.34 microg g(-1) DW) of total toxin burden, indicating that MCs had been transferred into offspring from their adults. Among the shrimp muscle samples analyzed, 31% were above the provisional WHO TDI level, suggesting the risk of consuming shrimps in Lake Chaohu. PMID- 15777959 TI - Bromoageliferin and dibromoageliferin, secondary metabolites from the marine sponge Agelas conifera, inhibit voltage-operated, but not store-operated calcium entry in PC12 cells. AB - Two alkaloids isolated from the marine sponge Agelas conifera were tested for interactions with cellular calcium homeostasis. Bromoageliferin and dibromoageliferin reduced voltage-dependent calcium entry in PC12 cells as measured with Fura II as calcium indicator. The half maximal concentration of both alkaloids to reduce voltage-dependent calcium entry was only slightly different: bromoageliferin showed a half maximal concentration of 6.61+/-0.33 microM, dibromoageliferin of 4.44+/-0.59 microM. Removal of calcium from extracellular solution for 10 min leads to an, at least, partial depletion of intracellular calcium stores, which induces a store-operated calcium entry after re-supplementation of calcium to the buffer. The store-operated calcium entry was unchanged by dibromoageliferin at a concentration of 30 microM, which fully blocks voltage-dependent calcium entry. The store-operated calcium entry induced by application of 5 microM thapsigargin was similarly not altered by 30 microM bromoageliferin. Both alkaloids reduce voltage-dependent calcium entry, but not store-operated calcium entry. The inhibition of voltage-operated calcium entry by bromoageliferin is shown in whole-cell patch clamp experiments. PMID- 15777960 TI - Morphology, toxin composition and pigment content of Prorocentrum lima strains isolated from a coastal lagoon in southern UK. AB - Prorocentrum lima was isolated from the coastal Fleet lagoon, Dorset, UK in 2000 and a number of clonal cultures established. These were analyzed for okadaic acid (OA), dinophysistoxin-1 (DTX-1), DTX-2, DTX-4 and diol esters by liquid chromatography coupled to mass spectrometry. OA concentrations varied from 0.4 to 17.1pg OAcell(-1) and DTX-1 from 0.4 to 11.3pg DTX-1cell(-1); DTX-2 was not detected in these isolates. OA and DTX-1 were detected in the culture media, as a result of toxin excretion. DTX-4 and a selection of DTX-4 diol esters were identified using selected ion monitoring, although not all strains produced these compounds. Cell size and number of marginal and valve pores of each strain were observed using scanning electron microscopy. OA and DTX-1 concentrations, pigment content and changes in nitrate and phosphate concentrations in the culture media were followed during growth of one strain of P. lima in batch culture. Diarrhetic shellfish poisoning (DSP) toxins have been previously detected in shellfish cultivated in the Fleet lagoon, but in the absence of any Dinophysis sp. cells. The identification of toxic P. lima strains from the Fleet suggests that this dinoflagellate is the most probable source of occasional DSP detected in the lagoon. PMID- 15777961 TI - A Lys49 phospholipase A(2) homologue from Bothrops asper snake venom induces proliferation, apoptosis and necrosis in a lymphoblastoid cell line. AB - Lys49 phospholipase A(2) homologues are abundant in viperid snake venoms. These proteins have substitutions at the calcium-binding loop and catalytic center which render them enzymatically inactive; however, they display a series of toxic activities, particularly cytotoxicity upon various cell lines in vitro. In this study we explored whether myotoxin II (MT-II), a Lys49 phospholipase A(2) homologue from the venom of the snake Bothrops asper, is capable of inducing various effects in a single cell type, using the lymphoblastoid B cell line CRL 8062 as a model. Cells were incubated with varying concentrations of MT-II for 24 and 48 h, time intervals that are more prolonged than the usual incubation times previously used in the characterization of this toxin. Results indicate that MT II induces proliferation at low concentrations (0.5-5.0 microg/mL). Apoptosis was predominant at higher toxin levels (5-25 microg/mL), whereas necrosis, associated with overt plasma membrane disruption, occurred at concentrations > or =25 microg/mL, and was the predominant effect at higher MT-II concentrations (50 microg/mL). It is concluded that a single phospholipase A(2) homologue can induce markedly different effects on a single cell line, depending on the concentration used, an observation that may have implications for the action of this type of venom component in vivo. PMID- 15777962 TI - Inhibition of tumor formation by snake venom disintegrin. AB - The metastasis of tumor cells to bone involves migration, invasion and adhesion to bone. Breast and prostate cancer cells have predilection for spreading to bone. Snake venom-derived arginine-glycine-aspartic acid (RGD)-containing disintegrins (e.g. rhodostomin) have been demonstrated to inhibit cell adhesion. Here, we found that rhodostomin inhibited the adhesion of breast and prostate carcinoma cells to both unmineralized and mineralized bone extracellular matrices in a dose-dependent manner, without affecting the viability of tumor cells. In addition, rhodostomin also inhibited the migration and invasion of breast and prostate carcinoma cells. It specifically inhibited the binding of monoclonal antibody (MoAb) 7E3, which recognizes integrin alphavbeta3, to tumor cells, but not those of other MoAbs against other integrin subunits such as alpha2, alpha3, alpha5 and beta1. As breast cancer cells MDA-MB-231 were locally injected into tibia in nude mice, histological examination of the tibia of control group revealed that most of the cancellous bone had been replaced by the breast cancer cells after 28 days' inoculation. In contrast, co-administration of trigramin with cancer cells markedly inhibited tumor growth and bone destruction. Taken together, disintegrins strongly inhibit the adhesion, migration, invasion of tumor cells and also tumor growth of human breast cancer cells in bone as well. Therefore, disintegrins may be developed as alternate therapy for bone metastasis of cancer cells. PMID- 15777963 TI - Inhibitory effect of phospholipase A(2) isolated from Crotalus durissus terrificus venom on macrophage function. AB - Recent work demonstrated that crotoxin, the main toxin of Crotalus durissus terrificus venom, inhibits macrophage spreading and phagocytic activities. The crotoxin molecule is composed of two subunits, an acidic non-toxic and non enzymatic polypeptide named crotapotin and a weakly toxic basic phospholipase A(2) (PLA(2)). In the present work, the active subunit responsible for the inhibitory effect of crotoxin on macrophage function was investigated. Peritoneal macrophages harvested from naive rats were used. Crotapotin (2.12, 3.75, or 8.37nM/ml), added for 2h to the medium of peritoneal cell incubation, did not modify the spreading and phagocytic activities of these cells. On the other hand, the PLA(2) (1.43, 2.86, or 6.43nM/ml) subunit caused a significant reduction (30, 33, and 35%, respectively) of the spreading activity. The PLA(2) also inhibited the phagocytosis of opsonised zymosan, opsonised sheep erythrocytes, and Candida albicans, indicating that this inhibitory effect is not dependent on the type of receptor involved in the phagocytosis process. The inhibitory effect of PLA(2) was not due to loss of cell membrane integrity, since macrophage viability was higher than 95%. These findings indicate that the inhibitory effect of crotoxin on macrophage spreading and phagocytic activities is caused by the phospholipase A(2) subunit. PMID- 15777964 TI - Antigenic relationships and relative immunogenicities of isolated metalloproteinases from Echis ocellatus venom. AB - The antigenic relationship between snake venom metalloproteinases (SVMPs) was analysed using rabbit antisera raised against the native forms of two SVMPs purified from Echis ocellatus venom. Using enzyme-linked immunosorbent assay (ELISA), western blotting and two-dimensional SDS-PAGE, our findings show that antibodies raised against EoVMP1, a non-haemorrhagic class P-I 24kDa SVMP, and EoVMP2, a haemorrhagic class P-III 56kDa SVMP, demonstrate cross-reactivities which relate to the domain hierarchy observed in class P-I to P-III/IV SVMPs. A third 65kDa P-III metalloproteinase (designated EoVMP3) was also isolated from E. ocellatus venom using hydrophobic interaction, size exclusion and anion exchange chromatography. In comparative immunoassays, EoVMP2 and EoVMP3 bound strongly to the commercial monovalent ovine Fab fragment antivenom EchiTAbtrade mark (raised against the same venom), but EoVMP1 showed no cross-reactivity. This could indicate that antivenoms may lack antibodies to potentially important venom components. PMID- 15777965 TI - Collection of venom from southern African scorpions. PMID- 15777966 TI - Lung cancer in women. AB - Lung cancer is one of the most important avoidable causes of death around the world, it is the most widespread carcinoma with a very poor prognosis, and is the leading cause of cancer death in both developed and developing countries. At present more men than women die each year from lung cancer, but in recent years a rapid increase in lung cancer mortality has been observed among women in developed countries, contrasting with a levelling off or decrease among men. The rising trend in female lung cancer mortality has been observed to parallel with the past and current prevalence of cigarette smoking among women in the United States and elsewhere. An important role of other factors acting either as independent risk factors or interacting with the effect of smoking has been suggested by some studies among women, among them genetic, biologic and hormonal factors, and probably some factors related to the environment and lifestyle. There is a controversy concerning the claim that women have a different susceptibility to tobacco carcinogens, which might or might not be greater than men do. Since tobacco is far and away the strongest epidemiological risk factor for the development of lung cancer, comprehensive smoking control efforts are the priority in the prevention of lung cancer among women. PMID- 15777967 TI - A common polymorphism in the 3'UTR of cyclooxygenase 2/prostaglandin synthase 2 gene and risk of lung cancer in a Chinese population. AB - Cyclooxygenases (COXs) are key enzymes that convert arachidonic acid to prostaglandins. Overexpression of COX-2, one of the COX isozymes, has been shown to be an early event in lung carcinogenesis and may play an important role in lung cancer development. A common single nucleotide polymorphism, T8473C, located within a potential functional region in the 3'UTR of COX-2 gene was identified and we hypothesized that this COX-2 variant is associated with lung cancer risk. To test this hypothesis, we genotyped this variant in a case-control study of 322 histologically-confirmed lung cancer patients and 323 age and sex frequency matched cancer-free controls in a Chinese population. The results showed that the frequencies of variant genotypes 8473CT/CC were significantly less common in the cases (27.3%) than in the controls (35.3%) (P=0.034), suggesting that the 8473C allele was protective against lung cancer. Multivariate logistic regression analyses revealed that the COX-2 variant genotypes (8473CT/CC) were associated with a significantly decreased risk of lung cancer compared with the 8473TT wild type homozygotes (OR=0.64, 95% CI=0.45-0.92). When we defined the reference group as non-smokers having the 8473CT/CC variant genotypes, the smokers with the 8473TT wild-type genotype had the greatest risk (adjusted OR=5.28, 95% CI=3.10 9.00). These findings indicate that the COX-2 T8473C polymorphism may contribute to lung cancer susceptibility in the Chinese population. Further larger molecular epidemiological studies are warranted to confirm these findings. PMID- 15777968 TI - Identification of intelectin overexpression in malignant pleural mesothelioma by serial analysis of gene expression (SAGE). AB - Malignant pleural mesothelioma (MPM) is a fatal neoplasm with no acceptable curative approaches. We used serial analysis of gene expression (SAGE) to compare the gene expression pattern of a surgically resected MPM to the autologous normal mesothelium. Intelectin gene overexpression (>139-fold) was found in the tumor. Online SAGE datasets revealed intelectin to be consistently present in mesothelioma(s), ovarian cancer, and colon cancer. Intelectin mRNA expression was found by RT-PCR in 4 of 5 resected MPM tumors, and Intelectin protein expression was confirmed by immunohistochemistry in 28 of 53 MPM tumors, and in 4 of 4 mesothelioma cell lines studied by Western blot. A marked induction in intelectin gene expression was observed among human primary mesothelial cells as a consequence of crocidolite asbestos exposure and simian virus 40 infection. Intelectin overexpression in mesothelioma could have potential screening, and therapeutic implications. PMID- 15777969 TI - Overexpression of vascular endothelial growth factor and its receptors in bronchial dypslasia demonstrated by quantitative RT-PCR analysis. AB - Neoangiogenesis is required for the growth of invasive lung carcinoma, however, the role of angiogenesis in the progression of premalignant changes to carcinoma of the lung is less clear. We have evaluated vascular endothelial growth factor (VEGF) expression and microvessel densities (MVDs) in 62 bronchoscopic biopsies from normal, reactive (basal cell hyperplasia (BCH)) and dysplastic bronchial epithelium and in tissue from twenty-seven invasive lung carcinomas in an effort to demonstrate angiogenic activity in these preneoplastic lesions and determine whether it is associated with increased bronchial epithelial VEGF expression. MVDs and VEGF RNA expression measured by quantitative RT-PCR were found to be elevated in comparison to normal bronchial tissue in bronchial dysplasias and invasive lung carcinomas but not in basal cell hyperplasias. Immunohistochemical (IHC) analyses revealed that expression of VEGF arose predominantly from bronchial epithelium. ELISA analysis of lung tumor tissue showed that elevated VEGF protein expression correlated with VEGF RNA levels (r=0.59, p=0.004). Increased expression of VEGF RNA was also found in histologically normal bronchial mucosa from patients with either dysplasia at other sites or a history of heavy tobacco use suggesting a possible field effect in regard to the elaboration of VEGF. Furthermore, analysis of VEGF isoforms and VEGF receptors by semi-quantitative RT-PCR in dysplastic and invasive lesions revealed characteristic altered patterns of expression in dysplasia and early cancer as compared to normal tissue. These results indicate that angiogenesis develops early in lung carcinogenesis and is associated with overexpression of VEGF. PMID- 15777970 TI - Cyclooxygenase-2 protein levels are independent of epidermal growth factor receptor expression or activation in operable non-small cell lung cancer. AB - Both cyclooxygenase (COX)-2 and epidermal growth factor receptor (EGFR) are thought to play important roles in the pathogenesis of non-small cell lung cancer (NSCLC). A number of in vitro studies have postulated a link between EGFR activation and subsequent COX-2 upregulation. The relationship between these factors has not been established in patients with NSCLC. COX-2 and EGFR expression were studied in 172 NSCLC specimens using standard immunohistochemical techniques. Western blotting was used to determine COX-2 and EGFR levels in five NSCLC cell lines. The effect of treatment with EGF on COX-2 expression in A549 cells was assessed. RESULTS: Both EGFR and COX-2 are overexpressed in NSCLC. The predominant pattern of COX-2 and EGFR staining was cytoplasmic. Membranous EGFR staining was seen in 23.3% of cases. There was no relationship between COX-2 and EGFR expression and survival or any clinicopathological features. No correlation was seen between EGFR expression and COX-2 expression in the immunohistochemical series or in the cell lines. Treatment with EGF did not upregulate COX-2 levels in A549 cells, either in serum free or serum-supplemented conditions. CONCLUSIONS: Although COX-2 and EGFR are over-expressed in NSCLC neither was of prognostic significance in this series of cases. There is no correlation between these two factors in either tumour samples or cell lines. Although these factors show no correlation in NSCLC, they remain potential, though independent targets for treatment. PMID- 15777971 TI - HER2/neu expression and amplification in non-small cell lung cancer prior to and after neoadjuvant therapy. AB - BACKGROUND: Expression and amplification of the HER2/neu protooncogene was analyzed in locally advanced NSCLC in a multimodality therapy approach in order to obtain information on the predictive value of HER2/neu for success or failure of neoadjuvant therapy. METHODS: In the scope of a prospective randomized phase III-trial, tumor tissue of pre-therapeutically obtained mediastinal lymph node biopsies (n=105) and corresponding post-surgical resection specimens (n=44) was analyzed by means of immunohistochemistry (DAKO-Hercep-Test) and fluorescence in situ hybridization (FISH). In 58 of 105 patients with metastatic mediastinal lymph node disease the extent of therapy-induced tumor regression could be established. RESULTS: Concerning HER2/neu expression, 16 lymph node biopsies (15.2%) showed 1+, 2+, or 3+ results. Five of these cases revealed amplification in FISH analysis (4.8%). In 44 corresponding resection specimens, Hercep-Test showed 1+, 2+, or 3+ results in 13 tumors (29.5%). Two of these patients revealed HER2/neu amplification in FISH analysis (4.5%). In patients with HER2/neu expressing tumors a trend towards a less extensive therapy-induced tumor regression could be demonstrated. When comparing pre-therapy and post-surgical results, there was a weak trend towards a selection of HER2/neu expressing tumor tissue in the course of neoadjuvant therapy. CONCLUSIONS: Only a limited subcollective of locally advanced NSCLC meets the biological requirements for anti-HER2/neu therapy. HER2/neu positive tumors appeared to be relatively resistant to chemotherapy and radiation treatment, none of these cases having a pathological complete or at least subtotal response in the corresponding resection specimens. This observation requires confirmation in large randomized controlled studies. PMID- 15777972 TI - Expression of E-cadherin and nm23 is associated with the clinicopathological factors of human non-small cell lung cancer in China. AB - E-cadherin, a calcium-dependent cell-cell adhesion molecule, functions as maintenance of epithelial integrity. nm23, encoded by non-metastatic 23 gene, plays a key role in differentiation of many kinds of epithelium. Loss or dysfunction of E-cadherin and nm23 was frequently identified in many types of human cancers and is considered to correlate with invasive/metastatic phenotype. We previously reported that defective expression of E-cadherin might play a role in the development of the malignant phenotype in non-small cell lung cancer (NSCLC) [Q.Y. Fei, H.T. Zhang, X.F. Chen, et al., Defected expression of Ecadherin in non-small cell lung cancer, Lung Cancer 37 (2002) 147-152]. In an attempt to evaluate the significance of E-cadherin and nm23 in human non-small cell lung cancer, we performed mRNA expression and genetic structure analyses of the E-cadherin and nm23 genes in 54 NSCLCs and 46 normal lung tissues. The mRNA expression was determined by semi-quantitative RT-PCR, and genetic structure was examined through PCR-SSCP followed by sequencing. Although no mutation of the E cadherin and nm23 genes was detected, the results obtained in the present study showed that reduction of E-cadherin and nm23 mRNA expression remarkably correlated with low histological differentiation, increasing stage as well as lymph node metastases (P<0.05). These data provide us with support for the idea that dysfunction of E-cadherin and nm23 has a role in progression of NSCLC and that the examination of E-cadherin and nm23 expression can provide experimental evidence for clinical treatment. PMID- 15777973 TI - Detection of plasma hnRNP B1 mRNA, a new cancer biomarker, in lung cancer patients by quantitative real-time polymerase chain reaction. AB - Circulating cell-free nucleic acids are noninvasive diagnostic tools for cancer detection. Heterogeneous nuclear ribonucleoprotein (hnRNP) B1, an RNA binding protein, has been found overexpressed in the early stage of lung cancer, including bronchial dysplasia, a premalignant lesion of lung squamous cell carcinoma. To determine the utility of plasma hnRNP B1 RNA and as cancer detection markers for lung cancer, we analyzed plasma hnRNP B1 mRNA of lung cancer patients by real-time RT-PCR. Plasma RNA was extracted from plasma of 44 lung cancer patients, 7 lung neoplasm patients, 24 benign lung diseases and 25 healthy volunteers. Mean concentration of plasma hnRNP B1 mRNA in lung cancer patients was 0.99 pg/microg RNA, whereas that in healthy volunteers and in benign lung diseases was 0.23 pg/microg RNA and 0.30 pg/microg RNA, respectively (p<0.05). Twenty of 44 (45.5%) lung cancer patients showed more than 0.70 pg/microg RNA of plasma hnRNP B1 mRNA, compared with only 3 of 25 (12.0%) healthy volunteers. Looking at histological subtype, squamous cell carcinoma patients showed higher hnRNP B1 mRNA in the plasma than did adenocarcinoma patients, which is consistent with our previous immunohistochemistry results. These results indicate that plasma hnRNP B1 mRNA is a useful non-invasive markers for detection of lung cancer. PMID- 15777974 TI - Endoscopic ultrasound guided biopsy versus mediastinoscopy for analysis of paratracheal and subcarinal lymph nodes in lung cancer staging. AB - BACKGROUND: Exact mediastinal evaluation of patients with non-small-cell lung cancer (NSCLC) is mandatory to improve selection of resectable and curable patients for surgery. Mediastinoscopy (MS) and endoscopic ultrasound guided fine needle aspiration biopsy (EUS-FNA) are considered complementary, MS covering the anterior- and EUS-FNA the posterior mediastinum. Both methods can reach the paratracheal- and subcarinal-regions, but little is known about which method is most accurate, when compared in patients having both procedures performed. The aim of this study was to assess and compare the diagnostic value of MS and EUS FNA with regard to mediastinal malignancy in the paratracheal- and subcarinal regions. METHODS: Sixty patients considered to be potential candidates for resection of verified or suspected NSCLC underwent MS and EUS-FNA. The EUS-FNA diagnoses were confirmed either by open thoracotomy, MS or clinical follow-up. RESULTS: MS and EUS-FNA were conclusive for paratracheal or subcarinal mediastinal disease in 6 and 24 patients, respectively. Two patients with N2 disease diagnosed by EUS-FNA were upstaged to N3 by MS. The sensitivity for lymph node metastases in the right paratracheal region (2/4R) was 67% for EUS-FNA versus 33% for MS (p=0.69). In the left paratracheal region (2/4L) the sensitivity of EUS-FNA was 80% versus 33% for MS (p=0.06). In the subcarinal region (7) the sensitivity of EUS-FNA was 100% versus 7% for MS (p<0.01). The sensitivity for lymph node metastases in region 2/4L and/or 2/4R and/or 7 was 96% for EUS-FNA versus 24% for MS (p<0.01). CONCLUSION: In our hands EUS-FNA was superior to MS in the examination of paratracheal- and subcarinal-regions of patients considered for resection of lung cancer. PMID- 15777975 TI - Patterns of care survey (PCS) in lung cancer: how well does current U.S. practice with chemotherapy in the non-metastatic setting follow the literature? AB - BACKGROUND: In LD-SCLC, combined modality therapy has emerged as the standard of practice in good performance status (PS) patients (pts). Pignon's meta-analysis [N Engl J Med 1992;327:1618-24] showed that combination chemotherapy (CT) and thoracic radiation (XRT) in LD-SCLC yielded an absolute 5.4% increase in 3-year survival versus chemotherapy alone. Concurrent chemoradiation upfront has generated the highest survival rates [Murray. J Clin Oncol 1993;11:336-44; Jeremic. J Clin Oncol 1996;15:893-900; Takada. J Clin Oncol 2002;20:3054-60]. In stage III NSCLC, multiple studies have shown therapeutic superiority for combination chemotherapy and XRT versus RT alone; and recent literature suggests a therapeutic advantage for concurrent chemoradiation versus chemotherapy followed by XRT [Curran. ASCO 2000;19:484a; Furuse. JCO 1999;17:2692-9; Zatloukal. ASCO 2002;A-1159]. Data are less secure regarding the role of chemotherapy in stage I and II NSCLC. MATERIAL AND METHODS: A stratified two-step cluster sampling technique was used for data collection. Five hundred and forty one individuals diagnosed between 1998 and 1999 with lung cancer, either LD-SCLC or stages I-III NSCLC were sampled from 58 institutions featuring radiotherapy facilities, giving a weighted sample size (wss) of 42,335 patients. All pts had Karnofski performance status (KPS) >or=60. We determined the percentage who received chemotherapy; the nature of chemotherapy and its timing with respect to XRT. SUDAAN statistical software was used to allow the incorporation of the design elements and weights to reflect the relative contribution of each institution and each patient in the analysis RESULTS: Of 72 (wss=6138) pts with LD-SCLC, 100% received XRT and 95% received chemotherapy (CT); 66% received concurrent (con) CT and XRT, of whom 29% also received CT pre XRT; 22% received CT post XRT as well, and 23% received both: 63% received sequential CT-->XRT+/ con CT; and 38% received some CT after XRT. Fifty-two percent received cisplatin (DDP), and 38% received carboplatin (CBDCA); 73% received etoposide (VP-16), while 10% received paclitaxel. Of 469 pts (wss=36,197) with NSCLC, 52% received CT, including 30% with stage I disease, 48% with stage II NSCLC, 60% with stage III NSCLC, and 50% with unknown stage. Thirty-nine percent received sequential CT ->XRT+/-CT, of whom 49% received CT pre XRT only. Seventy-four percent received con CT and XRT; and 27% received posterior CT, of whom 84% also received con CT/XRT. Forty-five received some CT in the pre-op setting and 15% in the post-op setting. Twelve percent received DDP-based therapy, while only 13% and 7% received VP-16 or vincas, respectively; 67% received CBDCA. Seventy-two percent received taxanes, of whom 96% received paclitaxel. Gemcitabine was administered to 3% of NSCLC pts. CONCLUSIONS: Combined modality therapy is typically employed in the therapy of LD-SCLC and LA-NSCLC. The majority of those treated for SCLC receive concurrent CT/XRT, while nearly 3/4 of those treated with CT and XRT for LA-NSCLC received concurrent CT/XRT. Current practice in the US generally matches evidence-based literature, although a significant percentage of practitioners substitute CBDCA for DDP in both venues and use paclitaxel in lieu of vincas or etoposide in NSCLC. PMID- 15777976 TI - Patterns of care survey (PCS) in chemotherapy (CHT) in nonmetastatic lung cancer. PMID- 15777977 TI - Stereotactic hypofractionated radiation therapy for stage I non-small cell lung cancer. AB - We reviewed our initial institutional experience with the use of stereotactic hypofractionated radiation therapy (SFRT) in patients with stage I non-small cell lung cancer (NSCLC). Thirty patients with inoperable stage I non-small cell lung cancer due to a severe chronic obstructive pulmonary disease (COPD) and/or chronic heart disease (Eastern Cooperative Oncology Group (ECOG) performance status of 0-2) were treated between December 2000 and October 2003 with SFRT in curative intent. Infiltration of locoregional lymph nodes and distant metastases were ruled out by computerized tomography (CT) scan of the brain, thorax, and abdomen, and by whole body FDG-positron emission tomography scan in all patients. Total RT doses ranged from 24.0 to 37.5 Gy, given in 3-5 fractions to the 60% isodose encompassing the planning target volume. Immobilization was carried out by a vacuum couch and a low-pressure foil. The clinical target volume was the tumor as it appeared in lung windowing on lung CT scan. Organ movements (caused by breathing; range, 6-22 mm) and reproducibility of patient positioning in the couch (range, 3-12 mm) were calculated by sequential CT and orthogonal films. The individual values were taken into account as a safety margin for the definition of the planning target volume (PTV). The median follow-up of living patients is 18 months (range, 6-38 months). As maximum response, there were 10 (33%) complete responses (CRs) and 14 (47%) partial responses (PRs), resulting in a total response rate of 80%. Stable disease was observed in 6 (20%) patients, while no patient experienced progressive disease. During follow-up, 2 (7%) local recurrences were observed (after 17 and 18 months, respectively). Of 5 (17%) patients who developed distant metastasis, 1 patient developed it in liver (3 months), another one in brain (6 months), and another one in the lung (36 months), while 2 patients developed it in mediastinal lymph nodes (after 8, and 11 months, respectively) only. Of 9 (30%) patients who have died, only 3 (10%) died of cancer, while 6 (20%) died of cancer-unrelated or unknown causes. Acute side effects were mild and affected 9 (33%) patients during the RT course (fatigue being the most frequent one in 6 patients). There were 22 acute events occurring in 19 (63%) patients during the first 3 months post-SFRT, the most frequent one being pneumonitis observed in 14 (46%) patients. However, there was only one (3%) grade 3 acute toxicity and no patient experienced greater than grade 3 toxicity during this study. One (3%) patient experienced rib fracture as the late event. SFRT is a feasible and safe treatment method in inoperable patients with stage I NSCLC having reduced lung capacity. Longer follow-up is necessary to get robust data on late toxicity as well as survival. PMID- 15777978 TI - Phase I study of combination chemotherapy with gemcitabine and irinotecan for non small cell lung cancer. AB - A phase I study combining a fixed dose of gemcitabine with differing doses of CPT 11 every two weeks for previously treated non-small cell lung cancer (NSCLC) patients. PATIENTS AND METHODS: A total of 21 patients with previously treated non-small cell lung cancer were treated every two weeks with CPT-11 followed by gemcitabine. The gemcitabine dose was fixed at 1000 mg/m2. The starting dose of CPT-11 (50 mg/m2) was then escalated different patients in 25 mg/m2 increments until 150 mg/m2 (level 5), the recommended dose as a single agent in Japan. RESULTS: Dose-limiting toxicity was only observed at level 5, in three of nine patients receiving the highest dose of CPT-11. One patient had grade 3 diarrhea, and two could not continue chemotherapy with grade 1 diarrhea or grade 1 neutropenia on day 15. Hematologic toxicity with this combination regimen, however, was generally mild. No grade 4 neutropenia, and only one case of grade 3 leukopenia was noted at level 5. Compliance with the combination regimen was good and there was no cumulative toxicity with the subsequent courses. Twenty-five courses of therapy were given at level 5 and the percentage of actual delivered doses/planned doses was 82%. CONCLUSIONS: The combination chemotherapy has only very mild toxicity and dose which can be recommended with this regimen are 1000 mg/m2 for gemcitabine and 150 mg/m2 for CPT-11 every two weeks. PMID- 15777979 TI - Increased dose-intensity of gemcitabine in advanced non small cell lung cancer (NSCLC): a multicenter phase II study in elderly patients from the "polmone toscano group" (POLTO). AB - Gemcitabine is usually administered at a planned dose-intensity (DI) from 750 to 800 mg/m2/week. Preclinical data have suggested a possible dose-response relationship of gemcitabine. A multicenter phase II study was conducted to evaluate the activity in terms of no progression rate (complete responses+partial responses+stable diseases) of gemcitabine administered at an increased DI (1000 mg/m2/week) in elderly advanced non-small-cell lung cancer (NSCLC) patients. Secondary endpoints were to evaluate tolerability, progression free survival and overall survival. Elderly (age>or=70 years) chemo-naive advanced NSCLC patients, ECOG PS 0-2, were treated with intravenous gemcitabine 1500 mg/m2 intravenous (30 min infusion) on days 1 and 8 every 21 days for four courses. One hundred and twenty-two patients with a median age of 75 years (range 70-84) entered the study. The following grade 3 (NCI-CTC) haematological toxicities were reported (percent of patients): neutropenia 2.4%, thrombocytopenia 1.6%, anaemia 2.4%. No grades 3-4 non-haematological toxicities were observed. Among 111 evaluable patients 52 (46.8%) no progressions, 17 (15.3%) partial responses (WHO criteria), 35 (31.5%) stable diseases and 59 (53.2%) progressions were observed. Median time to progression was 3.2 months and median duration of survival was 5.4 months. The overall 1-year survival rate was 27%. Although increased dose-intensity of gemcitabine in elderly NSCLC patients is feasible without severe toxicities, this does not seem to be associated with an increased activity and efficacy in comparison to standard gemcitabine regimens with lower dose-intensities. PMID- 15777980 TI - Phase II trial of oral vinorelbine in combination with cisplatin followed by consolidation therapy with oral vinorelbine in advanced NSCLC. AB - BACKGROUND: Among the cytotoxic agents commonly combined with cisplatin in the treatment of advanced NSCLC, vinorelbine has led to significant outcome improvements. Adding more than four cycles of the combination regimen increase toxicities. The availability of an oral form of vinorelbine appeared as a particularly convenient way to provide a consolidation treatment to patients who have achieved an objective response or stable disease. PATIENTS AND METHODS: This multi-centre phase II open-label, non-comparative study was designed to evaluate the treatment with four cycles of the combination chemotherapy with oral vinorelbine at the dose of 60 mg/m2 on day 1 and day 8 for the first cycle and then 80 mg/m2 plus cisplatin 80 mg/m2 on day 1 every 3 weeks followed for patients with objective response or stable disease by consolidation therapy with oral vinorelbine at 80 mg/m2 weekly on patients with unresectable localised or metastatic non-small-cell lung cancer (NSCLC). The primary endpoint was tumor response. The secondary objectives were progression free-survival, overall survival and toxicity assessment. Visual analogue scales (VAS) filled by the patients were also used to evaluate subjective changes under treatment, reflecting patients' clinical benefit. RESULTS: Fifty-six patients enrolled into the study from April 2001 to April 2002 received the combination regimen. Twenty five patients (43.9%) also received the subsequent consolidation treatment. Partial tumor responses were obtained in 13 patients (26.5%, 95% CI 15.0-41.1) of 49 evaluable patients. Stable disease was observed in 22 (44.9%) of patients. The median duration of response was 6 months (95% CI 4.3-8.2). The median progression free-survival was 4.2 months (95% CI 2.8-6). The median overall survival time was 10 months (95% CI 7.4-14) and the 1 year survival was 42.6%. The main toxicities recorded were haematological. Grade 3 and 4 neutropenia were observed in 16 patients (29.1%). Nausea, vomiting and fatigue were the major non-haematological toxicities reported. Among the symptoms recorded by the patients on VAS scales (appetite, fatigue, pain, cough, dyspnea, haemoptysis), except anorexia, all symptoms were improved during the combination therapy and in the consolidation phase. CONCLUSION: This study confirms that the efficacy of the cisplatin/oral vinorelbine combination in NSCLC is comparable to cisplatin/I.V. vinorelbine. This study also suggests that consolidation therapy with vinorelbine alone may probably prolong the efficacy of the combination regimen. The convenience offered to patients by an oral form of vinorelbine is a definite asset for consolidation therapy. PMID- 15777981 TI - Octreotide treatment for paraneoplastic intestinal pseudo-obstruction complicating SCLC. AB - Small cell lung cancer (SCLC) is often associated with paraneoplastic neurological syndromes like intestinal pseudo-obstruction. This syndrome is characterized by dysmotility of the bowel without mechanical obstruction. The pathogenesis of the syndrome is thought to involve autoimmune mechanisms with production of antineuronal antibodies and enteric neuronal degeneration. We report a patient with severe constipation as a clinical presentation of a paraneoplastic intestinal pseudo-obstruction complicating SCLC, who was successfully treated with the somatostatin analogue octreotide. This may be explained by effects of hormone-like substances from the tumor directly inhibiting the gut motility, rather than by autoimmune mechanisms. PMID- 15777982 TI - Efficacy of the tyrosine kinase inhibitor gefitinib in a patient with metastatic small cell lung cancer. AB - Gefitinib (Iressa, ZD1839) is an orally active inhibitor selective for the epidermal growth factor receptor tyrosine kinase and has shown promise in the treatment of non-small cell lung cancer (NSCLC). There has been no report to date of the effect of gefitinib treatment in patients with small-cell lung cancer (SCLC). Here we report a case of metastatic SCLC that was successfully treated with gefitinib. This case is the first reported of objective clinical response after gefitinib treatment in a patient with SCLC. Our report suggests that treatment with gefitinib is a novel option for a subset of patients with SCLC. PMID- 15777983 TI - Quantification of free RNA in serum and bronchial lavage: a new diagnostic tool in lung cancer detection? AB - Circulating cell-free nucleic acids have been detected in serum. In cancer patients levels of free DNA seem to be higher than in non-tumor controls and the detectable nucleic acids are partly of tumor origin. We asked whether free RNA can be detected as well in cell-free bronchial lavage fluid (BLF) supernatant, and whether quantification of free RNA allows to discriminate between tumor and non-tumor patients. 73 patients with lung cancer (NSCLC n=62, SCLC n=11) and 56 patients with non-malignant lung diseases were included. The RNA was isolated from 1 mL serum and lavage, respectively with the Quiamp MinElute Virus Vacuum Kit (Qiagen, Germany). A real-time quantitative RT-PCR assay was used for transcript quantification of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene. Intact RNA was detectable in cell-free supernatants of BLF from 126/129 patients and was investigated in all 64 serum samples. RNA levels were higher in the cell-free supernatant of BLF than in serum. RNA concentration in the BLF from tumor patients was higher than in patients with a benign lung disease (p=0.009). In conclusion, quantification of intact RNA isolated from BLF supernatant and from serum might become a valuable tool for differentiation between tumor and non tumor patients. PMID- 15777984 TI - Letter to the editor. PMID- 15777985 TI - Cut-off levels of CYFRA21-1 to differentiate between metastatic and non metastatic NSCLC. PMID- 15777987 TI - Antigenic proteins associated with calcareous corpuscules of taenia solium: partial characterization of a calcium-binding protein. AB - BACKGROUND: A protein fraction was isolated from calcareous corpuscles of Taenia solium cysticerci. The antigens in this fraction were recognized in ELISA and Western blot assays by all sera from a group of patients with active neurocysticercosis (NC) and were not recognized by the sera from patients with other neurological disorders. Western blot analysis also showed that several high molecular weight proteins were strongly recognized by antibodies in all the neurocysticercotic patient sera, suggesting a potential for serological diagnosis of neurocysticercosis. METHODS: In order to characterize these antigenic proteins, we used a monoclonal antibody raised against a high MW calcium-binding protein associated with calcareous corpuscles of Echinococcus granulosus (EgCaBP1). RESULTS: Western blot assays revealed the recognition of a protein band of about 260 kDa, appearing within the range of the high MW antigens recognized by the NC sera. Several cDNA clones were isolated through screening of a T. solium metacestode library with a DNA probe for EgCaBP1, containing partial coding sequences showing about 88% identity with the protein of E. granulosus. Moreover, a recombinant product expressed in bacteria from the partial coding sequence of T. solium showed the ability to bind Ca2+ and was recognized by the monoclonal antibody. This recombinant calcium-binding protein of T. solium was not recognized by the NC patient sera by ELISA and Western blot. CONCLUSIONS: Antigenic proteins in the calcareous corpuscles of T. solium metacestodes deserve further analysis as candidates in the development of diagnostic tools for neurocysticercosis. PMID- 15777988 TI - Mixture of honey, beeswax and olive oil inhibits growth of Staphylococcus aureus and Candida albicans. AB - BACKGROUND: Honey, beeswax and olive oil mixture (1:1:1, v/v) is useful in the treatment of diaper dermatitis, psoriasis and eczema. The study was designed to investigate effects of honey, olive oil, and beeswax and the mixture on growth of Staphylococcus aureus and Candida albicans isolated from human specimens. METHODS: The following experiments were performed: 1) honey mixture was poured on holes made on plates seeded with S. aureus or C. albicans, 2) the microorganisms were cultured onto media made of honey mixture alone, nutrient agar-honey mixture and Sabouraud glucose agar-honey mixture. The concentration of honey mixture in nutrient agar or Sabouraud glucose agar was 12.5, 25, 33, 50 and 66% (v/v), and 3) honey, olive oil or beeswax was added onto nutrient agar or Sabouraud glucose agar at a ratio of 1:2 (v/v) and then were seeded with S. aureus or C. albicans. RESULTS: Clear zone of inhibition was observed around holes filled with honey mixture; 3.5 mm on media seeded with C. albicans and 4 mm on media seeded with S. aureus. No growth of either microorganism was obtained on media made of honey mixture alone. The minimum concentration of honey mixture in nutrient agar-honey mixture media required to inhibit S. aureus was 50% and 66% concentration was required to inhibit C. albicans growth onto Sabouraud glucose agar-honey mixture media. No growth of S. aureus or C. albicans was obtained on media containing honey whereas mild to moderate growth was obtained on media containing olive oil or beeswax. CONCLUSIONS: Honey and honey mixture apparently could inhibit growth of S. aureus or C. albicans. PMID- 15777989 TI - Serum anti-Yersinia antibody in Chinese patients with Kawasaki disease. AB - BACKGROUND: Many infectious agents have been implicated as an etiology to develop Kawasaki disease (KD). In Taiwan, studies on the relationship between Yersinia and KD have not been reported. METHODS: We measured sera for anti-Yersinia antibodies by using enzyme immunoassay (EIA) in 31 patients with KD and 60 healthy children (HC). Yersinia strains included Y. pseudotuberculosis I, II, III, IV, V, VI and Y. enterocolitica O3, O8 and O9. RESULTS: Data of 31 patients with KD showed that for the IgG antibody, serum anti-Y. pseudotuberculosis II, III, Y. O8 and O9 antibody were significantly higher when compared to the HC. Except for Y. pseudotuberculosis IV, all other Yersinia strains of either IgA or IgM antibodies increased significantly in patients with KD vs. the HC. If we compared the number of patients who had significant elevation of OD and those of HC, we found IgA anti-Yersinia antibodies (PST I, PST II, O3, O8, O9), IgM (PST VI, O8) and IgG (PST II, O8, O9) were significantly elevated in KD patients than in HC. A significant relationship was present between KD with myocarditis and increased anti-Yersinia antibody titer. CONCLUSIONS: The findings in this study suggest that preceding Yersinia infection may play a role in the pathogenesis of KD. Further study of the relationship between KD with myocarditis and increased anti-Yersinia antibody is needed. PMID- 15777990 TI - Polymorphisms related to estrogen and xenobiotic metabolism in healthy Turkish women. AB - BACKGROUND: Polymorphisms which are inherited alterations in the activity of cytochrome P450 1B1 (CYP1B1), catechol O-methyltransferase (COMT), manganese superoxide dismutase (MnSOD) hold the potential to define differences in estrogen metabolism and, thereby, possibly explain inter-individual differences in cancer susceptibility associated with estrogen-mediated carcinogenesis. METHODS: The CYP1B1 (L432V), COMT (V158M), MnSOD (Ala-9Val) genotypes, to examine estrogen metabolism and the influence of age of menarche/menopause, were determined by using different polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) based on genotyping assays. RESULTS: Women who carried CYP1B1 *3 and COMT-L alleles had an earlier age at menarche than the women who carried wild alleles (chi2 = 4.57, p = 0.032), whereas I did not observe any correlation in women with all mutant alleles. Also, CYP1B1 *3 and COMT-H genotypes were common among postmenopausal women with a body mass index (BMI) > 27 kg/m2 (Fisher exact test, p = 0.044). CONCLUSIONS: To my knowledge, this is the first genetic study on the association of these genes with susceptibility in Turkish women. Although the small sample size of each combination of estrogen metabolizing, results suggest that the CYP1B1 *3 and COMT-L alleles influence age at menarche in healthy Turkish women. PMID- 15777991 TI - Clinical profile of pediatric HIV infection from India. AB - BACKGROUND: Our aim was to study the clinical profile of pediatric patients admitted with HIV infection. METHODS: The prospective study was conducted from January 2000 to October 2001 at a tertiary care referral teaching hospital in Mumbai, India. Admitted in-patients (aged 1 month to 12 years) detected to be HIV positive (on triple ELISA test) were enrolled in the study. HIV status of patients < 18 months of age was confirmed by DNA-PCR testing. Demographic data, clinical features, investigations and outcome were recorded in a pre-designed proforma. RESULTS: Fifty HIV-positive children (31 males and 19 females; M:F = 1.6:1) were enrolled. Thirty cases were completely immunized, 9 were partially immunized while 11 were not immunized. Forty-two were perinatally infected, while eight cases were infected via blood transfusion (patients with thalassemia major on chronic transfusion therapy). Clinical features at presentation in 42 symptomatic cases included protein-energy malnutrition (90%), fever > 1 month (50%), weight loss > 1 month (50%), persistent generalized lymphadenopathy (24%) and skin manifestations (79%). The gastrointestinal (62%) and respiratory (52%) were the most commonly involved organ systems. Opportunistic infections noted included tuberculosis (19 cases), candidiasis (6 cases), Pneumocystis carinii pneumonia (4 cases), herpes zoster (3 cases) and giardiasis (1 case). Six patients died (mortality, 14%). CONCLUSIONS: Perinatal transmission is the most common mode of acquiring HIV in the pediatric age group. Most patients have protein-energy malnutrition. Tuberculosis is common in HIV-infected Indian children. Patients with HIV-encephalopathy have a poor outcome. PMID- 15777992 TI - Renal handling of uric acid in patients with type 1 diabetes in relation to glycemic control. AB - BACKGROUND: The aim of the study was to compare the renal handling of uric acid (UA) in 16 patients with type 1 diabetes without renal failure (age 34.8 +/- 13.3 years) and in 15 healthy subjects (age 34.9 +/- 12.6 years). METHODS: Creatinine clearance (Cr-Cl), clearance of uric acid (UA-Cl), fractional excretion of uric acid (UA-FE), and 24-h urinary UA excretion (UA-U) were determined. Glycemic control was assessed using fasting glucose, glycated hemoglobin and fructosamine tests. RESULTS: Patients with diabetes had significantly (p < 0.0001) lower serum UA concentrations compared to control group (2.8 +/- 0.7 vs. 5.7 +/- 0.8 mg/dl), and higher urinary UA excretion (813 +/- 107 vs. 423 +/- 40 mg/day), UA clearance (21.9 +/- 7.1 vs. 5.2 +/- 0.9 mL/min) and fractional UA excretion (17.1 +/- 5.5 vs. 4.8 +/- 1.3%), with higher creatinine clearance (129 +/- 16 vs. 111 +/- 12 mL/min, p < 0.005). In patients with diabetes there was a strong negative correlation between serum UA concentration and UA 24-h excretion (R = -0.79; p < 0.001). Fractional UA excretion correlated with fasting glycemia and HbA1c (R = +0.51 and +0.53; p < 0.05). CONCLUSIONS: In type 1 diabetes there is significant UA renal clearance increase, which is higher with poor glycemic control. It leads to hypouricemia despite an approximately twofold UA excretion increase and therefore despite increased UA synthesis. PMID- 15777993 TI - Hypomagnesemia after major abdominal operations in cancer patients: clinical implications. AB - BACKGROUND: Serum magnesium levels are rarely measured in routine chemistry panels. The extent and impact of postoperative serum magnesium changes remain unclear. METHODS: One hundred seventy-one cancer patients who had undergone celiotomy procedures during a 38-month period were evaluated retrospectively for postoperative electrolyte alterations, with special emphasis on serum magnesium. Clinicopathologic predictors and early postoperative outcome correlations were examined. RESULTS: There were 151 major procedures and 20 minor operations. All postoperative electrolyte and hematocrit values were significantly different from preoperative values, except for serum phosphate. Preoperative total serum magnesium (normal range: 1.7-2.5 mg/dL {0.7-1.03 mmol/L}), obtained prior to any bowel cleansing, differed from postoperative levels (means +/- standard deviation: 2.0 +/- 0.46 vs. 1.53 +/- 0.33 mg/dL; p < 0.0001). A lowered postoperative serum magnesium was observed in those patients who had either undergone an operation with curative intent (p = 0.0035), a major resection (vs. no resection, p = 0.0259), or preoperative bowel cleansing with sodium phosphate (p = 0.024). Other laboratory serum parameters that correlated with the postoperative magnesium level included postoperative levels of phosphate (p = 0.009), potassium (p = 0.01), and total calcium (p = 0.012), as well as preoperative calcium (p = 0.017). The complication rate was 20%, with five postoperative deaths (2.9%). Postoperative morbidity was predicted by preoperative potassium (p = 0.004) and albumin levels (p = 0.016); deaths were predicted by postoperative infections (p = 0.0007) and correlated to postoperative hypokalemia (p = 0.03). CONCLUSIONS: Major abdominal cancer operations lead to significant electrolyte alterations. The severity of these changes correlates with the resection extent, especially in procedures with curative intent. In addition, bowel cleansing with sodium phosphate may participate in lowering serum magnesium as well as other electrolytes. In light of our postoperative magnesium replacement policy, no untoward events could be linked to postoperative hypomagnesemia in this series. To evaluate the impact of postoperative hypomagnesemia or magnesium replacement on postoperative outcomes requires a prospective randomized trial. PMID- 15777994 TI - Oral infections and glycemic control in pregnant type 2 diabetics. AB - BACKGROUND: The relationship between diabetes mellitus and infection is bidirectional. Diabetes favors infection, while infections make controlling diabetes much more difficult. The most frequent infections encountered in pregnant diabetics are those of the urinary tract (UTI) and cervicovaginal (CVI) area. Periodontal diseases (PD) and active caries (AC) are infectious diseases frequently found in these cases often remaining as hidden entities of low intensity. The aim of this study was to assess whether there is an association between PD and caries with lack of glycemic control often encountered in pregnant type 2 diabetic women. METHODS: A single skilled researcher performed the periodontal evaluation of PD and AC cases. Glycated alpha hemoglobin (HbA(1c)) over 6.5% was used to diagnose metabolic glucose disturbances. The controlled variables were UTI, CVI and adherence to treatment. The statistical tests used were chi-square adjusted for continuity, analysis of variance, odds ratio with 95% confidence intervals and logistic regression with a significance level of p < 0.05. RESULTS: One hundred twenty-seven type 2 diabetic women were seen during the second trimester of their pregnancies. Mean age was 31.3 years; 55% had lack of glycemic control associated with CVI (chi2 21.07, p < 0.000), PD (chi2 5.72, p < 0.005) and UTI (chi2 13.77, p < 0.000) with therapeutic adherence (TA) (chi2 14.80, p < 0.00). No association was found with AC. The logistic regression results showed that UTI, CIV, PD and TA are associated with lack of glycemic control. CONCLUSIONS: Periodontal diseases may act as "hidden" infections in pregnant diabetics and be associated with lack of glycemic control. This situation should be taken into consideration by healthcare teams in charge of prenatal care. PMID- 15777995 TI - Estimation of population pharmacokinetic parameters of free-phenytoin in adult epileptic patients. AB - BACKGROUND: Serum concentrations of free-phenytoin (F-PHT) obtained in adult epileptic patients receiving PHT in monotherapy were analyzed to estimate the Michaelis-Menten pharmacokinetic parameters. METHODS: Steady-state F-PHT serum concentrations, PHT dosing history, and associated information were collected prospectively. The maximum metabolic rate (Vm) and Michaelis-Menten constant (Km) of F-PHT and their interindividual variability data were estimated using nonlinear mixed effects modeling (NONMEM). RESULTS: Twenty-nine patients with two or more available steady-state F-PHT serum concentrations (total of 63 dose/serum concentration pairs) met the inclusion criteria. Patients were taking PHT (100 500 mg/day) in monotherapy. The population estimates of F-PHT for Vm and Km were 9.1 mg/kg/day and 7.3 mg/L, respectively. The model was prospectively evaluated in a small group (seven) of additional patients. CONCLUSIONS: The recommended daily dose in this population to achieve a F-PHT concentration of 1.5 mg/L is 6.1 mg/kg. PMID- 15777996 TI - Association between systemic lupus erythematosus, rheumatoid arthritis, hyperprolactinemia and thyroid autoantibodies. AB - BACKGROUND: Hyperprolactinemia (hyperPRL) has been associated with autoimmune rheumatic disorders and the presence of thyroid autoantibodies (tAb). The interrelation between these variables was the focus of this prospective study. METHODS: The study assessed six groups of individuals: 26 with systemic lupus erythematosus (SLE), 20 with rheumatoid arthritis (RA), 28 with tAb (tAb+), 14 with untreated hyperprolactinemia (hyperPRL), 10 with treated hyperPRL, and a control group (n = 28). Prolactin (PRL), free thyroxin, TSH, antibodies against thyroglobulin (TgAb), thyroid microsomal antigen (MsAb) and/or thyroid peroxidase (TPOAb) were determined in all patients. Those with hyperPRL had macroprolactin investigated by the polyethylene glycol (PEG) precipitation method. RESULTS: PRL (ng/mL) levels in the SLE, RA, and tAb+ groups were, respectively, 21.3 +/- 12.6, 11.5 +/- 7.4, and 12.5 +/- 8.6, and were significantly greater in the SLE group (p = 0.006) than in the controls (12.5 +/- 6.5) and in the other groups. Five patients had hyperPRL: three with SLE, one with RA, and one with tAb+. Macroprolactinemia was detected in three of the untreated hyperprolactinemic patients and in the hyperprolactinemic patient of the tAb+ group. Positivity for any of the tAb was 15% in the SLE, 15% in the RA, 57.1% in the untreated hyperPRL, 10% in the hyperPRL on treatment, and 3.6% in the control group. The presence of antibodies was significantly more frequent in the untreated hyperPRL group than in the control group (p = 0.001). CONCLUSIONS: The results indicate that the PRL level is higher in SLE patients and that in the presence of hyperPRL there is increased prevalence of antithyroid antibodies, evidencing the association of PRL and autoimmunity and pointing to the appropriateness of assessing and monitoring the progress of these markers in patients affected by these disorders. PMID- 15777997 TI - Increased triglyceride levels in a Japanese population living in southern Brazil. AB - BACKGROUND: In the present study we investigated 96 individuals of Japanese descent living in southern Brazil (Cascavel-PR) in terms of triglyceride (TG) levels (> or < or = 200 mg/dL) and compared them to non-Japanese control individuals. METHODS: We analyzed TG and total cholesterol (TC) levels by an enzymatic method and apolipoprotein A-I and B (apo A-I and apo B) by a turbidimetric method. We also determined the lipoproteins HDL and LDL by a direct method and by electrophoresis. All these determinations were performed in plasma. RESULTS: TG levels were above 200 mg/dL in 18.7% of the individuals of Japanese descent and in 8.4% of the controls. Mean TC levels were 259 mg/dL for Japanese descendants and 225 mg/dL for the control group. We observed that individuals of Japanese descent with TG levels above 200 mg/dL had the highest TC, LDL-c, and VLDL-c levels and the lowest HDL-c and apo A-I levels. Body mass index (BMI) was also higher in individuals of Japanese descent with TG above 200 mg/dL. CONCLUSIONS: This Japanese population has high TG levels compared to control individuals, and diet did not influence these levels. PMID- 15777998 TI - Recent trends in mortality due to chronic obstructive pulmonary disease (COPD) in Mexico, 1980-2002. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterized by airflow limitation that is not fully reversible. Changes in classification have a major impact on reported mortality rates. METHODS: Between 1980 and 2002, 230,463 COPD cases were studied (age group 35-94 years); 134,579 men; 95,884 women. RESULTS: The crude mortality rate varied from 37 (1980) to 61 per 100,000 men (2002), (increase: 65%). The crude mortality rate for females increased from 27 (1980) to 43 deaths per 100,000 women (2002), (increase: 56%). The trends of the absolute values by birth year and age groups are higher as age increases. As age of death comes down and the birth cohort increases, the absolute values decrease. The Mexican states located in the northern and central areas present a higher risk for dying. There is an increment coefficient of 93 cases per increment year in males (age group 35-74), and 61 cases per increment year in females (age group 35-74 years). For the age group 35-94 years, the annual increase for males is 288 cases. These results were statistically significant, and the regression model was validated by residual analysis. CONCLUSIONS: The oldest cohorts of the studied population showed the highest COPD mortality absolute values. The geographic risk of dying from COPD is concentrated in two regions: a) the three Mexican states of higher economic income at the northern frontier to the U.S. and b) those Mexican states surrounding the main producer of tobacco (Nayarit). PMID- 15777999 TI - A new test of disk-diffusion to help in staphylococci identification. AB - A modified carbohydrate fermentation test (MCFT) of disk-diffusion assay was performed using a peptone-free agar medium, in order to improve the identification of clinical staphylococci species. Results were compared with the standard method of carbohydrate fermentation in tubes (SCFT) containing methyl red broth. MCFT was able to identify correctly 12 different species of staphylococci from the ATCC collection and 646 clinical isolates. MCFT showed sensitivity between 91.6% and 100% and specificity between 99.6% and 100%, for the different carbohydrates studied. MCFT could replace the SCFT, with the evident advantage of reducing time for materials preparation. The method presented herein is easy to perform in clinical or research microbiology laboratories, because it can differentiate species of staphylococci. PMID- 15778000 TI - Effect of zinc-treated Entamoeba histolytica on the human polymorphonuclear respiratory burst. AB - One of the mechanisms that Entamoeba histolytica uses to evade host immune response is inhibition of the polymorphonuclear (PMN) leukocyte respiratory burst. In studies previously conducted in a model used in our laboratory, we observed that when treating trophozoites with different zinc concentrations certain amebic functions are inhibited while significantly limiting development of hepatic abscess in golden hamsters (Mesocricetus aureatus). We carried out an in vitro study using a chemoluminescent method to assess the effect zinc-treated amebic trophozoites exercise on respiratory burst in human PMNs. We measured response of PMNs incubated with E. histolytica trophozoites from cultures with TYI-S33 medium alone and with zinc. Zinc concentrations between 0.1 and 1.0 mM did not affect amebic trophozoite viability, and PMNs in contact with these in a zinc-free medium had an oxidative response similar to that obtained with zymosan and significantly greater (p <0.05) than that generated by cells co-incubated with amebas cultured in TYI-S33 medium alone. These results suggest that zinc alters the amebic mechanism that inhibits the oxidative function of human polymorphonuclear leukocyte. PMID- 15778001 TI - Nasopharyngeal colonization by Moraxella catarrhalis and study of antimicrobial susceptibility in healthy children from Cuban day-care centers. AB - The prevalence of nasopharyngeal carriage of Moraxella catarrhalis was determined for the first time in Cuba. One-hundred fifty healthy children attending three day-care centers in the municipality of Marianao, Havana City were studied. The percentage of recovering bacteria in nasal and pharyngeal swabs was compared. Antimicrobial susceptibilities to ampicillin, trimethoprim-sulfamethoxazole, tetracycline, cefotaxime, ceftriaxone, chloramphenicol, erythromycin, azithromycin, amoxicillin/clavulanate, and norfloxacin were determined by the disk diffusion method according to recommendations of the National Committee for Clinical Laboratory Standards. Sixty-five percent of the children studied carried Moraxella catarrhalis. The nasal cavity was the main isolation site for this organism (81% of positive cultures). Most strains were highly susceptible to the antimicrobial agents tested, except to ampicillin (53.6% resistance). This study provides evidence of the need for continued surveillance of antimicrobial susceptibility of Moraxella catarrhali, in order to determine optimal empiric therapy for community-acquired respiratory tract infections produced by this pathogen. PMID- 15778002 TI - Dynamic profiling of estrogen receptor and epidermal growth factor signaling in the uteri of genistein- and estrogen-treated rats. AB - The pharmacokinetics and time course actions of the soy isoflavone, genistein, and estradiol benzoate (EB) on sex steroid and growth factor signaling were compared in the rat uterus. Following one s.c. injection of 500 mg genistein/kg BW or 500 microg EB/kg BW, AUC for genistein was 20171.8 ng h/ml and was 15.7 ng h/ml for estradiol-17-beta. Estrogen receptor-alpha (ER-alpha) decreased within 2 h of genistein or EB treatment, returning to basal levels within 24 and 48 h, respectively. In response to genistein and EB, progesterone receptors (PRs) A and B increased between 16 and 24 h, with a significant increase at 24 and 48 h. Epidermal growth factor receptor (EGFR) expression peaked 16 h after genistein or EB treatment, inversely correlating with extracellular regulating kinase (ERK) phosphorylation. These effects were inhibited by antiestrogen pretreatment, demonstrating a requirement for ER. At 16 h, uterine weight, epithelial cell height, and cell proliferation increased. While EGFR levels increased, phosphorylated EGFR was not altered. Reduced phosphorylation of downstream kinases corresponded with decreased stromal phosphorylated-ERK (P-ERK) immunolabeling, suggesting signal attenuation. Dynamic profiling of sex steroid receptors and EGF signaling molecules suggest a similar mechanism of action for genistein and EB in the uterus, albeit at approximately 1000-fold concentration. PMID- 15778003 TI - Human erythrocytes are affected by the organochloride insecticide chlordane. AB - Chlordane is a widely used organochlorine insecticide. In order to evaluate its perturbing effect upon the morphology of human erythrocytes it was caused to interact with human red cells and molecular models of cell membranes. These consisted in bilayers of dimyristoylphosphatidylethanolamine (DMPE) and of dimyristoylphosphatidylcholine (DMPC), representative of phospholipid classes located in the inner and outer monolayers of the erythrocyte membrane, respectively. Scanning electron microscopy (SEM) observations indicated that this pesticide induced a significant alteration in the shape of the erythrocytes as they changed their discoid shape to spherocytes. According to the bilayer couple hypothesis, the shape changes induced in erythrocytes by foreign molecules are due to differential expansion of their two monolayers. The fact that chlordane produced spherocytes would indicate that the pesticide was equally located in the outer and the inner moieties of the red cell membrane. This conclusion was supported by the results obtained from X-ray diffraction studies. These showed that the hydrophobic and polar head regions of DMPC bilayers were perturbed when the insecticide was in a 1:10 molar ratio with respect to the lipid. These results were confirmed by the fluorescence experiments performed in DMPC large unilamellar vesicles (LUV). Chlordane produced a sharp decrease in the anisotropy and general polarization parameters in the 0-0.1 mM range, implying an increase in the fluidity at the acyl chain and polar region of DMPC. On the other hand, the bilayer structure of DMPE was perturbed in a fashion similar to that observed by X-ray diffraction in DMPC, a fact that explains the morphological change induced by chlordane to the human erythrocytes. PMID- 15778004 TI - Polycyclic aromatic hydrocarbons, carbon monoxide, "tar", and nicotine in the mainstream smoke aerosol of the narghile water pipe. AB - A smoking machine protocol and yields for "tar", nicotine, PAH, and CO are presented for the standard 171-puff steady periodic smoking regimen proposed by Shihadeh et al. [Shihadeh, A., Azar, S., Antonios, C., Haddad, A., 2004b. Towards a topographical model of narghile water-pipe cafe smoking: A pilot study in a high socioeconomic status neighborhood of Beirut, Lebanon. Pharmacology Biochemistry and Behavior 79(1), 75]. Results show that smokers are likely exposed to more "tar" and nicotine than previously thought, and that pyronsynthesized PAH are present in the "tar" despite the low temperatures characteristic of the tobacco in narghile smoking. With a smoking regimen consisting of 171 puffs each of 0.53l volume and 2.6s duration with a 17 s interpuff interval, the following results were obtained for a single smoking session of 10 g of mo'assel tobacco paste with 1.5 quick-lighting charcoal disks applied to the narghile head: 2.94 mg nicotine, 802 mg "tar", 145 mg CO, and relative to the smoke of a single cigarette, greater quantities of chrysene, phenanthrene, and fluoranthene. Anthracene and pyrene were also identified but not quantified. The results indicate that narghile smoke likely contains an abundance of several of the chemicals thought to be causal factors in the elevated incidence of cancer, cardiovascular disease and addiction in cigarette smokers. PMID- 15778005 TI - Effect of betaine supplementation on changes in hepatic metabolism of sulfur containing amino acids and experimental cholestasis induced by alpha naphthylisothiocyanate. AB - Alterations in the hepatic metabolism of sulfur amino acids in experimental cholestasis induced by alpha-naphthylisothiocyanate (ANIT) (100 mg/kg, po) were monitored in male mice for 1 week. We also examined the effects of betaine supplementation (1% in drinking water) for 2 weeks on the hepatotoxicity and changes in the sulfur amino acid metabolism induced by ANIT treatment. Acute ANIT challenge elevated the serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST) activities, and total bilirubin contents from 5 h after the treatment, reaching a peak at t = 48-72 h. Hepatic S adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) levels were decreased significantly in a manner almost inversely proportional to the changes in serum parameters measured to determine the ANIT-induced toxicity. Hepatic glutathione and cysteine levels were elevated at t = 120 h after the treatment. Betaine supplementation blocked or significantly attenuated induction of the hepatotoxicity by ANIT. The decrease in SAM and SAH levels was also inhibited by betaine intake. The results indicate that betaine supplementation may antagonize the induction of experimental cholestasis and changes in the metabolism of sulfur amino acids associated with ANIT treatment. The underlying mechanism and pharmacological significance of its action are discussed. PMID- 15778006 TI - Human exposure to dioxins from food, 1999-2002. AB - In response to aggressive attempts to control dioxin emissions over the last 35 years, human exposures to dioxins from the environment have declined significantly. The primary source of human exposure to dioxins at present is food. The sources of dioxins in food are not well understood and are probably varied. Data on the levels of dioxins measured in various foods for samples collected from 2000 to 2002 have recently been released by the US Food and Drug Administration as part of its Total Diet Study. Data on samples collected in 1999, and released in 2002, are also available. Based on those data and on the US Department of Agriculture's most recent food consumption survey (1994-1996 & 1998 Continuing Survey of Food Intakes by Individuals), estimates of dioxin intake for the total US population and for three age groups of children were obtained. Results show that the most recent mean dietary exposures for all groups are below 2 pg TEQ/kg BW/day, the tolerable daily intake established for dioxins by the World Health Organization. Between 1999 and 2002 mean dioxin intakes from food appear to have decreased, but when estimates are adjusted based on a standardized limit of detection and evaluating only those {congenerxfood} combinations common to all 4 years, no trend is apparent. When dioxin concentrations below the limit of detection are represented by one-half the limit, approximately 5% of the intake estimates for 2-year-olds and 1% of the intake estimates for 6-year-olds exceed the tolerable daily intake by about 10%, although such upper-percentile estimates should not be equated with excess risk. When non-detectable dioxin values are set to zero (i.e., when only dioxin values actually measured are used), only 1% of intake estimates exceed the tolerable daily intake for 2-year olds. As expected, about 50% of daily dietary dioxin intake by the total US population is attributable to meat and dairy products, based on the same food group classifications used by the National Academy of Sciences' Committee on the Implications of Dioxin in the Food Supply. This information may be useful for targeting future risk management activities. PMID- 15778007 TI - Percutaneous penetration of diethanolamine through human skin in vitro: application from cosmetic vehicles. AB - Concern has been raised over the safety of diethanolamine (DEA) which may be present as a minor component of alkanolamide ingredients of cosmetic formulations. Skin penetration data were therefore generated for a range of typical formulations under in-use conditions. Seven rinse-off formulations (A-E, G and H), a leave-on emulsion (F), representing prototype cosmetic formulations and containing representative levels of DEA were prepared. Target levels of DEA were attained by inclusion of DEA as either (14)C-DEA or a combination of (14)C DEA and unlabeled DEA. Skin permeation and distribution were evaluated using human skin in vitro, static diffusion cells and phosphate buffered saline (pH 7.4) as the receptor phase. At least 12 replicate epidermal membranes were prepared from a minimum of four donors for each test group. Receptor phase samples were taken at appropriate time intervals. At the end of the test period, radioactivity remaining on the skin surface and on the diffusion cell donor cap was determined before the skin samples were tape-stripped. The remaining tissue was solubilized and radioactivity determined. Permeation was very low from all vehicles applied under in-use conditions (range 1-48 ng/cm(2) over 24 h). Comparison was also made between permeation and distribution of DEA from an infinite dose of a simple aqueous solution and the leave-on formulation (F) through paired samples of fresh and frozen full thickness skin from the same donors. When applied as an infinite dose in aqueous solution DEA permeation at 24 h was greater through frozen than through fresh skin. From the leave-on formulation, permeation was similar and very low for both fresh and frozen skin. Recovery of DEA after application of the aqueous solution to fresh human skin and subsequent aqueous and organic extraction of the epidermal and dermal tissue indicated that the majority (>98%) of DEA was in the aqueous extract, suggesting that DEA was in the free state and not associated with the lipid fraction. These data provide a basis for the estimation of the potential systemic exposure and safety margins for DEA in representative cosmetic formulations. PMID- 15778008 TI - Fumonisins in foods from Cordoba (Argentina), presence and genotoxicity. AB - Fumonisins B(1), B(2)yB(3) (FB(1), FB(2)yFB(3)), are a group of toxins produced by different mold species, Fusarium moniliforme and Fusarium proliferatum being the most important ones. Its compounds were tested in chromosome aberrations (CA), sister chromatid exchange (SCE), and micronucleus (MN) in human lymphocytes, and, in Allium cepa (onion), the chromosomal aberrations (CA) assay was used. Moreover, the presence of fumonisins and their producer moulds was determined in different food substrata in Cordoba city, Argentina. Cytogenetic studies using FB(1), FB(2) and FB(3) levels gave positive results for the higher concentrations (5 and 10mug/g) with FB(1). As regards the cytogenetic aspect of FB(1), we found an increase in the incidence of genetic damage measured by chromosomal aberrations, sister chromatid exchange, micronuclei and chromosomal aberrations in Allium cepa. These results indicate that human lymphocytes cells and plants cells (Allium cepa) have a very sensitive cellular response to the mycotoxin fumonisin B(1) as observed at the highest concentrations. PMID- 15778009 TI - 4-Week repeated intravenous dose toxicity study of a new camptothecin anticancer agent CKD-602 in dogs. AB - CKD-602 is a new camptothecin derivative antitumor agent with a formula (7-[2-(N isopropylamino)ethyl]-(20S)-camptothecin) developed by Chong Kun Dang Pharmaceutical Company in Korea. In the present study, the subacute toxicity of CKD-602 was investigated after 4-week repeated intravenous administration of the test chemical in beagle dogs. The test chemical was administered intravenously at dose levels of 0, 0.001, 0.005, or 0.01 mg/kg/day for 4 weeks to male and female dogs (n = 3 for male and female dogs for each dose). During the test period, clinical signs, mortality, body weights, food and water consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross findings, organ weights and histopathology were examined. In the high dose group, an increase in the incidence of abnormal clinical signs and a decrease in food and water intake and body weight gain were observed in both sexes. Hematological investigations revealed decreased white blood cells (WBC) in both sexes and reduced red blood cells (RBC), hemoglobin and hematocrit in females. Histopathological examinations revealed an increase in the incidence of atrophy of the sternal and femoral marrow and spleen in both sexes and atrophy of the thymus and mesenteric lymph node in males. No treatment-related adverse effects were observed in both sexes of the low and middle dose groups. In conclusion, the 4-week repeated intravenous dose of CKD-602 to beagle dogs caused increases in the clinical signs and histopathological changes, and decreases in the body weight gain, food and water intake, RBC, hemoglobin, hematocrit and WBC at the dose level of 0.01 mg/kg/day. In the present experimental conditions, the target organs were determined to be bone marrow, blood cells, spleen, thymus, and mesenteric lymph node. The no observed-adverse-effect levels (NOAEL) for males and females were considered to be 0.005 mg/kg/day, respectively. PMID- 15778010 TI - Inhibition of human cytochrome P450 enzymes by the natural hepatotoxin safrole. AB - The hepatotoxin, safrole is a methylenedioxy phenyl compound, found in sassafras oil and certain other essential oils. Recombinant cytochrome P450 (CYP, P450) and human liver microsomes were studied to investigate the selective inhibitory effects of safrole on human P450 enzymes and the mechanisms of action. Using Escherichia coli-expressed human P450, our results demonstrated that safrole was a non-selective inhibitor of CYP1A2, CYP2A6, CYP2D6, CYP2E1, and CYP3A4 in the IC(50) order CYP2E1 < CYP1A2 < CYP2A6 < CYP3A4 < CYP2D6. Safrole strongly inhibited CYP1A2, CYP2A6, and CYP2E1 activities with IC(50) values less than 20 microM. Safrole caused competitive, non-competitive, and non-competitive inhibition of CYP1A2, CYP2A6 and CYP2E1 activities, respectively. The inhibitor constants were in the order CYP1A2 < CYP2E1 < CYP2A6. In human liver microsomes, 50 microM safrole strongly inhibited 7-ethoxyresorufin O-deethylation, coumarin hydroxylation, and chlorzoxazone hydroxylation activities. These results revealed that safrole was a potent inhibitor of human CYP1A2, CYP2A6, and CYP2E1. With relatively less potency, CYP2D6 and CYP3A4 were also inhibited. PMID- 15778011 TI - Promoting insulin secretion in pancreatic islets by means of bisphenol A and nonylphenol via intracellular estrogen receptors. AB - In this study, we investigated the effects of endocrine disrupters bisphenol A (BPA) and nonylphenol (NP) on insulin secretion from rat pancreatic islets. Following acute exposure to BPA and NP, neither BPA nor NP (0.1, 1, 10, 100 and 1000 microg/l) affected insulin secretion in concentrations of 16.7 mM glucose. However, insulin secretion following long-term exposure to BPA or NP for 24 h in 16.7 mM glucose was significantly higher than without exposure. To determine whether increased insulin secretion resulting from long-term exposure to BPA and NP is induced via intracellular estrogen receptors, we blocked the cytosolic/nuclear estrogen receptors, using actinomycin-D (Act-D), an inhibitor of RNA synthesis, and ICI 182,780 (ICI), an estrogen receptor inhibitor. Following long-term exposure to BPA (10 microg/l) or NP (10 microg/l), Act-D or ICI treatment eliminated the facilitation of insulin secretion. In conclusion, we have demonstrated for the first time that long-term exposure to endocrine disrupters, such as BPA and NP, promotes in vitro insulin secretion from the pancreatic islets, via cytosolic/nuclear estrogen receptors. PMID- 15778012 TI - Role of cyclooxygenase-2 in deoxynivalenol-induced immunoglobulin a nephropathy. AB - Ingestion of the trichothecene mycotoxin deoxynivalenol (DON) induces serum IgA elevation and kidney mesangial IgA deposition in a manner that mimics the early stages of IgA nephropathy (IgAN), the most common human glomerulonephritis. Previous studies indicate that elevated interleukin-6 (IL-6) expression is crucial for this model and that DON induction of cyclooxygenase-2 (COX-2) might drive IL-6 upregulation. We hypothesized that COX-2 and its metabolites are essential for DON-induced IgAN and thus might be a suitable target for prophylaxis against aberrant IgA upregulation. DON feeding studies using COX-2 knockout mice or the COX-2 specific inhibitor, rofecoxib (Vioxx), were employed to test the hypothesis. Study 1 results demonstrated that DON consumption induced serum IgA and IgA-immune complex (IC) accumulation, IgA kidney deposition and splenic IgA secretion in wild-type mice. COX-2 deficiency did not affect upregulation of these parameters but rather, promoted DON-induced serum IgA elevation. Study 2 demonstrated that rofecoxib could not block DON-induced serum IgA, serum IgA-IC and mesangial IgA accumulation but instead increased enhanced serum IgA upregulation. These corroborating results suggest that COX-2 is not a requisite for DON-induced IgAN and furthermore, that COX-2 inhibitors such as rofecoxib would be contraindicated for the prevention of early stages of IgAN. PMID- 15778013 TI - Chronic toxicity and carcinogenicity of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin displays a distinct dose/time toxicity threshold (c x t = k) and a life prolonging subthreshold effect. AB - Chronic toxicity of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) including its carcinogenicity was studied in female Sprague-Dawley rats in lifetime experiments. Six single dose and three multiple dose rate experiments were conducted with a single dose corn oil control group and a multiple dose rate corn oil control group, respectively. The lowest dose (1.0 mg/kg) of HpCDD and multiple dose rates of corn oil (4.0 ml/kg every other week) both prolonged the life of rats by about 2 months over that of single dose corn oil controls. Higher doses resulted in a predictable shortening of the life of rats after single dose administrations as well as after multiple dose rate administrations. The c x t = k paradigm previously validated for acute toxicity [Toxicol. Sci. 49 (1999) 102] was confirmed for chronic toxicity including carcinogenicity of HpCDD. The c x t = k product was independent of dosing regimen. Anemia and squamous cell carcinoma of the lungs were the earliest and most prevalent endpoints of toxicity. A dose of 2.1 mg/kg and 3.1 mg/kg of HpCDD caused 16.6% and 73.3% lung cancer, respectively. Liver cancer had a low prevalence and was a very late effect occurring only at doses lethal acutely for most rats in the three highest dosage groups. There was no correlation in the dose-dependence of non-malignant hepatic lesions and liver cancer. PMID- 15778014 TI - Bran extracts from pigmented rice seeds inhibit tumor promotion in lymphoblastoid B cells by phorbol ester. AB - Using flow cytometry, we quantitatively evaluated anti-tumor-promoting activity of rice bran extracts by measuring inhibition of Epstein-Barr virus early-antigen activation (EBV-EA) induced by the tumor promoter 12-O-tetradecanoylphorbol-13 acetate (TPA). This assay measures anti-tumor-promoting activity and cytotoxicity of target substances using the same batch of cells. Using this short-term procedure, we have determined the anti-tumor-promoting activity of 70% ethanol water extracts of bran (outer layer) from seeds of five pigmented rice cultivars: Jumlalocal-1, DZ 78, Elwee, LK1-3-6-12-1-1, and LK1A-2-12-1-1. The results show that, compared to an extract from the non-pigmented white cooking rice variety Chuchung, the extracts from the pigmented varieties strongly inhibited phorbol ester-induced tumor promotion in marmoset lymphoblastoid cells B95-8 in vitro. These findings may facilitate development and use of new health-promoting rice varieties. PMID- 15778015 TI - Hypolipidemic and antioxidant properties of tocotrienol rich fraction isolated from rice bran oil in experimentally induced hyperlipidemic rats. AB - We investigated a dose-dependent hypolipidemic and antioxidant effect of tocotrienol rich fraction (TRF) isolated from rice bran oil on experimentally induced hyperlipidemic rats. Feeding of atherogenic diet (5% hydrogenated fat, 0.5% cholic acid and 1% cholesterol) for three weeks resulted in a significant increase in plasma triglyceride (3.3-fold) and total cholesterol (2.4-fold) levels. There was a 5-fold increase in the level of LDL cholesterol with only a small increase in HDL cholesterol. On the other hand, HMG-CoA reductase activity was significantly reduced in these animals. The formation of TBARS, thiobarbituric acid reactive substances, (86%) and conjugated dienes (78%) were also significantly higher in these rats compared to normals. After the induction of hyperlipidemia for three weeks, rats were supplemented with different doses of TRF for one week. TRF supplementation decreased the lipid parameters in a dose dependent manner with an optimum effect at a dose of 8 mg TRF/kg/day. HMG-CoA reductase activity, which was increased after the withdrawal of atherogenic diet, remained significantly decreased during the TRF treatment. Feeding of TRF also decreased TBARS and conjugated dienes significantly. These results suggest that TRF supplementation has significant health benefits through the modulation of physiological functions that include various atherogenic lipid profiles and antioxidants in hypercholesterolemia. PMID- 15778016 TI - Cytotoxicity of Fusarium mycotoxins to mammalian cell cultures as determined by the MTT bioassay. AB - Fusarium mycotoxins occur worldwide in cereal grains and animal feeds and cause outbreaks of Fusarium mycotoxicoses in humans and animals. In this study mammalian cell cultures were used to screen the cytotoxicity of the most common Fusarium mycotoxins; deoxynivalenol (DON), zearalenone (ZEN), fumonisin B(1) (FB(1)) and moniliformin (MON). The most sensitive cell line for each Fusarium mycotoxin was determined for further toxicological investigations as an alternative to whole animal testing. Chinese hamster ovary cells (CHO-K1) were found to be the most sensitive for DON and FB(1) with IC(50) values of 0.27 and 85.5 microg/ml, respectively, after 48-h exposure. The hepatocellular carcinoma cells (HepG2) showed the highest sensitivity to MON with IC(50) values of 39.5 for 48 h and 26.8 microg/ml for 72-h exposure. Balb/c mice keratinocyte cell line (C5-O) was found to be the most sensitive to ZEN with IC(50) of 24.1 microg/ml after 72-h exposure. DON was found the most cytotoxic to the cell cultures of all the mycotoxins tested, followed by MON, ZEN, and FB(1). The results indicated that CHO-K1, C5-O, and HepG2 cells were found to be the sensitive cell lines for preliminary screening of DON, ZEN and MON contaminated feed and food extracts, respectively. PMID- 15778017 TI - Dietary modulation of 7,12-dimethylbenz[a]anthracene (DMBA)-induced adrenal toxicity in female Sprague-Dawley rats. AB - In this study, dietary modulation of 7,12-dimethylbenz[a]anthracene (DMBA) induced adrenal toxicity in rats was investigated. Beginning at postnatal day (PND) 21, female Sprague-Dawley rats were fed either soy-containing NIH-31 diet or soy- and alfalfa-free 5K96 diet. On the first day of diestrus when the animals were PND 50 +/- 5, rats received either an oral dose of 80 mg/kg DMBA or sesame oil, the vehicle, and were sacrificed at 24, 36, or 48 h after treatment. Apoptosis was manifested at 24 and 36 h after DMBA treatment in the zona reticularis (ZR) and the zona fasciculata (ZF) of the adrenal cortex; this was followed by severe hemorrhagic necrosis at 48 h. DMBA-induced apoptosis, evaluated by the TUNEL assay, immunohistochemical analysis of activated caspase 3, and the ratio of expression of pro-apoptotic Bax to anti-apoptotic Bcl2, was greater in rats fed NIH-31 diet relative to rats fed 5K96 diet at 24 h after treatment. Four of six DMBA-treated rats fed 5K96 diet had severe adrenal necrosis by 48 h, whereas this lesion was present in only two of six DMBA-treated rats fed NIH-31 diet. DMBA also caused a significant decrease of serum corticosterone relative to controls at 48 h in rats fed 5K96 diet. The present study indicated that diet modulates DMBA-induced adrenal toxicity in female rats, with increased apoptosis early and reduced necrosis later in rats fed a soy containing diet. PMID- 15778018 TI - The effect of pharmacologically altered gastric pH on cadmium absorption from the diet and its accumulation in murine tissues. AB - Solubility of Cd in Cd-amended mouse chow in water was reduced by increased pH; even less Cd was solubilized by simulated digestion in vitro, where increased gastric phase pH decreased solubility, an effect that persisted following intestinal digestion at pH 5.5. These data suggested that increasing gastric pH in vivo pharmacologically would reduce Cd accumulation in target organs of mice treated with omeprazole (a proton-pump inhibitor) or cimetidine (a H2-receptor antagonist). This expectation was mostly not realized. Gastric pH in animals receiving Cd-amended diet was increased by omeprazole, but not cimetidine, relative to animals receiving no drugs, and Cd-amended diet. Tissue concentrations of Cd were similar among the three groups receiving Cd-amended diet, for liver, kidney and testes. Small intestine Cd concentration was lower for omeprazole-treated animals than for those receiving neither drug and Cd amended diet, suggesting that omeprazole decreased Cd absorption by this organ. This effect may have been compensated for by increased uptake of complexed Cd by the large intestine, as accumulation in the liver, kidney and testes was not reduced. In vitro determinations of bioaccessible Cd in food may not predict in vivo bioaccumulation in all target organs. PMID- 15778019 TI - Toxicology and safety of anti-oxidant of bamboo leaves. Part 1: Acute and subchronic toxicity studies on anti-oxidant of bamboo leaves. AB - The anti-oxidant of bamboo leaves (AOB) has recently been certificated as a novel kind of natural anti-oxidant by the Ministry of Health of the People's Republic of China, and has been used in various food systems. Here, AOB was subjected to a series of acute and subchronic toxicological tests to evaluate its safety. It was examined to evaluate acute oral toxicity by using Kun-Ming mice and Sprague Dawley rats, and its mutagenic potential assessed by reverse mutation test using Salmonella typhimurium, bone marrow cell micronucleus test using Kun-Ming mice, and sperm abnormality test using Kun-Ming mice. In addition, a 90-day oral toxicity study using Sprague-Dawley rats was conducted to evaluate subchronic toxicology. The results showed that the maximum tolerated dose (MTD) of AOB was >10 g/kg body weight in both rats and in mice, which can be regarded as virtually non-toxic. No mutagenicity evidence was detected in any of the three mutagenic tests. Administration at levels of 1.43, 2.87 and 4.30 g/kg per day to the rats for 90 days did not induce significant hematological, clinic, chemical and histopathological changes, and suggested a no-observed-adverse-effect level (NOAEL) of 4.30 g/kg per day. These results indicate that AOB can be generally regarded as safe for use as a food additive. PMID- 15778020 TI - Long-term combined administration of quercetin and daidzein inhibits quercetin induced suppression of glutathione antioxidant defenses. AB - In this study, we investigated the effects of long-term administration of quercetin with or without daidzein on glutathione and the enzymes involved in its metabolism in rat liver in vivo. Male Sprague-Dawley rats were divided randomly into four groups and given oral quercetin (20 mg/day) and daidzein (20 mg/day) alone or in combination, or vehicle alone for six weeks. The serum and liver alpha-tocopherol concentrations were significantly increased following administration of quercetin and daidzein alone or in combination. Glutathione concentration and glutathione reductase activity was significantly (p < 0.05) decreased with quercetin treatment, while no such effect was observed with daidzein treatment. Interestingly, decrease in glutathione concentration and glutathione reductase activity by quercetin treatment was inhibited by combined administration of daidzein and quercetin. The malondialdehyde concentration was significantly decreased following administration of quercetin and daidzein alone or in combination. These results suggest that quercetin, but not daidzein, acts as a pro-oxidant agent by decreasing glutathione concentration and glutathione reductase activity. Interestingly, this pro-oxidant effect of quercetin was inhibited by the combined administration of quercetin and daidzein. PMID- 15778021 TI - Bluetongue virus in the French Island of Reunion. AB - This paper records the results of a bluetongue virus (BTV) serological survey and reports the first isolation of BTV on the French Island of Reunion. In January 2003, the French Island of Reunion, located off the coast of Madagascar, reported an outbreak of disease in cattle that resembled clinical bluetongue (BT) in sheep. The suspected causal agent was isolated and identified as epizootic haemorrhagic disease of deer virus (EHDV). However, because of the similarity in the clinical signs to those of BT, a retrospective survey against BTV was carried out using sera collected in 2002. Results revealed the presence of antibody in all sera tested indicating that BTV has been resident on the Island since 2002, and probably earlier. Although up to July 2003 no clinical BT had ever been reported in sheep, BTV viral RNA was amplified by RT-PCR from a single sheep blood collected in February that year, which strongly suggested that BTV was currently circulating on the Island. Following a second outbreak of disease in August 2003, this time involving a flock of Merino sheep, infectious BTV was finally isolated, and identified by both traditional and molecular techniques as serotype 3. The nucleotide and amino-acid sequences of the RT-PCR products amplified for BTV segments 7 and 10 from the sheep blood collected in February and August from different areas of the Island, were sufficiently diverse as to suggest that they were of different origins and/or different BTV serotypes. PMID- 15778022 TI - Genotyping of the fish rhabdovirus, viral haemorrhagic septicaemia virus, by restriction fragment length polymorphisms. AB - The aim of this study was to develop a standardized molecular assay that used limited resources and equipment for routine genotyping of isolates of the fish rhabdovirus, viral haemorrhagic septicaemia virus (VHSV). Computer generated restriction maps, based on 62 unique full-length (1524 nt) sequences of the VHSV glycoprotein (G) gene, were used to predict restriction fragment length polymorphism (RFLP) patterns that were subsequently grouped and compared with a phylogenetic analysis of the G-gene sequences of the same set of isolates. Digestion of PCR amplicons from the full-length G-gene by a set of three restriction enzymes was predicted to accurately enable the assignment of the VHSV isolates into the four major genotypes discovered to date. Further sub-typing of the isolates into the recently described sub-lineages of genotype I was possible by applying three additional enzymes. Experimental evaluation of the method consisted of three steps: (i) RT-PCR amplification of the G-gene of VHSV isolates using purified viral RNA as template, (ii) digestion of the PCR products with a panel of restriction endonucleases and (iii) interpretation of the resulting RFLP profiles. The RFLP analysis was shown to approximate the level of genetic discrimination obtained by other, more labour-intensive, molecular techniques such as the ribonuclease protection assay or sequence analysis. In addition, 37 previously uncharacterised isolates from diverse sources were assigned to specific genotypes. While the assay was able to distinguish between marine and continental isolates of VHSV, the differences did not correlate with the pathogenicity of the isolates. PMID- 15778023 TI - Genetic and antigenic characterization of the matrix protein of two genetically distinct ovine lentiviruses. AB - Small Ruminant Lentiviruses (SRLV) are a group of non-oncogenic retroviruses including Maedi-Visna virus (MVV) and Caprine Arthritis-Encephalitis virus (CAEV), which cause a chronic, multisystemic disease in sheep and goats, respectively. Phylogenetic analyses of SRLV are based in most cases on partial pol sequences. Several reports indicate that the species specificity of these viruses is not as strict as previously thought; MVV-like viruses have been found in goat populations and vice versa. Recently, the sequencing of some Italian ovine isolates has shown the presence of a new cluster more similar to classical caprine isolates (CAEV-like). Few data are available on the variability of structural proteins involved in the antibody response of infected animals. In this study, the gag gene of two genetically distinct ovine isolates, namely the MVV-like It-561 and the CAEV-like It-Pi1, was sequenced and the epitopes of matrix protein (MA) were mapped. Recombinant MAs and their subunits from both ovine aforementioned strains were tested against a panel of sheep and goat sera. Reactive epitopes were found in all three subunits of MA, although the central subunit displayed a more consistent reactivity. Epitope mapping of this subunit demonstrated that the amino acid sequence of at least one immunodominant epitope was quite different in the two strains. This antigenic variability may affect the sensitivity of a single strain-based immunoassay and suggests that both SRLV genotypes should be used in the development of future diagnostic tests, to avoid viral strain selection during the eradication programmes. PMID- 15778024 TI - Phylogenetic analysis of classical swine fever virus isolated from Taiwan. AB - By analyzing the E2 sequences of classical swine fever virus from field outbreaks in Taiwan during 1993-2001, three virus populations with distinct genotypes were determined including one historical (subgroup 3.4) and two exotic (subgroup 2.1) strains. The first subgroup 2.1 virus was isolated in 1994 and further sporadic outbreaks occurred after 1996. Phylogenetic analysis using the E2 region has segregated the Taiwanese strains of 2.1 virus into two different genotypes (termed 2.1a and 2.1b). The 2.1b viruses were only isolated in 2001 and shared approximately 94.8% nucleotide identities to the 2.1a viruses in the total genomic sequences. The results suggest that the 2.1a and 2.1b viruses may be introduced from different origins. PMID- 15778025 TI - A novel genetic marker to differentiate feline herpesvirus type 1 field isolates. AB - Five recent field isolates of feline herpesvirus type 1 (FHV-1) were compared by digestion with a restriction endonuclease, SalI or MluI. The SalI digestion showed a potentially useful difference in one isolate 00-035 that had an approximately 3.0 kbp fragment instead of a 2.6 kbp fragment in the other strains. After cloning the 3.0 and 2.6 kbp fragments, the nucleotide sequences were analyzed. The result showed that the 3.0 kbp fragment of 00-035 included a complete open reading frame of the herpes simplex virus 1 (HSV-1) homologue of the UL17 gene and a 5'-part of UL16 gene and that only one nucleotide substitution was found in the 5'-region of UL17 gene where the SalI site of the 2.6 kbp fragment locates. Based on these nucleotide sequences, two PCR primers were designed to amplify the region around the SalI site in the UL17 gene and the PCR was carried out using 78 field isolates from various parts of Japan. The SalI digestion of the PCR products revealed an interesting profile in that the genotype without the SalI site in UL17 gene was dominant in Tottori and Yamagata prefectures (69% and 75%, respectively) but minor in the other regions of Japan (0-10%). These results suggest that the SalI digestion method described in the present study can be used as a genetic marker to differentiate some FHV-1 field isolates and this is the first report that showed different distributions of FHV 1 genotypes using the novel genetic marker. PMID- 15778026 TI - Characterization of Env antigenicity of feline foamy virus (FeFV) using FeFV infected cat sera and a monoclonal antibody. AB - To characterize neutralizing antigenicity in relation to env genotypes of feline foamy virus (FeFV), serological analyses were performed using FeFV-infected cat sera and several field isolates including two env genotypes (F17- and FUV-types). Since three cats from which FeFV were isolated were found to have undetectable titers of virus neutralization (VN) antibodies, even to the homologous virus, VN antibodies were further examined with complement supplementation as an enhancement factor. With the presence of complement, the VN titers of FeFV infected cat sera increased drastically. Although most of serum samples neutralized strains of either env genotype, sera sampled from two cats neutralized all the strains examined at similar titers, suggesting that superinfection with both env genotypes of FeFV might have occurred in the two cats. Further, we produced a monoclonal antibody (mAb) specifically neutralizing FeFV strains of FUV-type. The mAb was shown to have higher affinity to an epitope on Env of FUV-type than that of F17-type by immunoprecipitation assay. This study supplies basic information important for studies on FeFV vector development as well as on the relationship between the virus and the host immune response. PMID- 15778027 TI - Mixed infections in vitro with different Chlamydiaceae strains and a cell culture adapted porcine epidemic diarrhea virus. AB - Assuming a synergistic or additive effect of Chlamydiaceae in coexistence with other enteropathogenic agents, the viral/bacterial interaction between a cell culture adapted porcine epidemic diarrhea virus (ca-PEDV) and different Chlamydiaceae strains was studied in vitro. Vero cells were dually infected with ca-PEDV and one of the three chlamydial strains Chlamydia trachomatis S45, Chlamydophila abortus S26/3 or Chlamydophila pecorum 1710S. Three experimental protocols were designed varying the inoculation sequence. Cell layers were first inoculated with Chlamydiaceae and 20 h later with ca-PEDV in protocol one. In protocol two, both agents were administered concurrently, whereas in protocol three, ca-PEDV was applied 20 h in advance of the Chlamydiaceae. Immunofluorescence techniques, immunohistochemical (IH) staining and electron microscopy were subsequently employed to investigate the cell layers. Using indirect immunofluorescence (IF) labeling, all mixed infections revealed dually infected cells, however, only incidentally and in low numbers. Characteristically, ca-PEDV syncytia with one or more chlamydial inclusions were detected but dually infected single cells were absent. Some syncytial cells contained enlarged C. abortus or C. pecorum inclusions with abnormally large developmental forms. In comparison with simultaneously conducted monoinfections, larger chlamydial inclusions were observed in dually infected cell layers. Experiments with C. trachomatis showed significantly increased numbers of chlamydial inclusions in dually infected cell layers compared to monoinfected ones. These findings indicate an influence of ca-PEDV on the chlamydial developmental cycle and in the case of C. trachomatis, a positive effect on chlamydial colonization in mixed infections. PMID- 15778028 TI - Comparison of vancomycin-resistant enterococci isolates from human, poultry and pigs in Korea. AB - Vancomycin-resistant enterococci (VRE) have emerged as an important nosocomial pathogen. Since 1989, a rapid increase in the incidence of enterococcal bacteremia and endocarditis by VRE has been reported. The use of avoparcin in animal husbandry is reportedly associated with the appearance of VRE. In this study, a multiplex polymerase chain reaction (PCR) method was established to detect and differentiate resistant types of enterococci, which specifically amplify the four van genes that encode vancomycin resistance elements. Using this method, we investigated the incidence rates and types of VRE from two types of farms: those that had used avoparcin and those that had not used avoparcin. A total of 1091 animal fecal samples were collected from 70 pig farms and 32 poultry farms. A total of 425 enterococci were isolated from the fecal samples. Among the 425 isolates, six showed a pattern of high-level vancomycin resistance (Minimal Inhibitory Concentration, MIC: 64-256 microg/ml). Out of six high-level VRE, three were isolated from poultry farms that had used avoparcin and three were not. The six high-level VRE harbored the vanA gene. Sixty-seven of 425 isolates that showed a pattern of low-level vancomycin resistance (MIC: 4-8 microg/ml) were associated with the presence of vanC-1 or vanC-2/3 gene. We also performed a repetitive extragenic palindromic PCR (rep-PCR) method to compare the genetic relatedness between the high-level VRE of six animal isolates and 31 human isolates. None of the animal isolates had a similar rep-PCR pattern as the human isolates but similarities between human VRE isolates were observed. PMID- 15778029 TI - Nasal carriage of Staphylococcus aureus in dairy sheep. AB - The purpose of this study was to assess the prevalence of Staphylococcus aureus nasal carriage of dairy sheep in farms producing cheeses manufactured with raw ewe's milk. The study showed that 29% of ewes carried S. aureus in their nares. The genetic diversity of the 136 isolates recovered from the anterior nares of the ewes, from the ambient air of the milking parlour and from cheeses was investigated using pulsed-field gel electrophoresis (PFGE) of DNA SmaI digests. The genotyping results showed that 75 out of 106 isolates recovered from nasal carriage in dairy sheep belonged to a dominant pattern (previously named OV) and a genetically related pattern (named OV'). The same profile (OV or OV') was found in the ambient air and cheeses, suggesting a continuum between isolates within these different compartments. PMID- 15778030 TI - Characterization of Escherichia coli O86 O-antigen gene cluster and identification of O86-specific genes. AB - Escherichia coli O86 belongs to the enteropathogenic E. coli (EPEC) group, some strains of which are pathogens of humans, wild birds and farm animals. The O antigen gene cluster of E. coli O86 was amplified by long-range PCR using primers based on the housekeeping genes galF and gnd, and then sequenced. Genes involved in GDP-Fuc and N-acetyl-galactosamine (GalNAc) synthesis and genes encoding glycosyltransferases, O-unit flippase and O-antigen polymerase were identified on the basis of homology. By screening against 186 E. coli and Shigella-type strains, two genes specific to E. coli O86 were identified. A polymerase chain reaction (PCR) assay, based on the specific O-antigen genes identified here, could be used for the rapid detection of E. coli O86 in environmental and clinical samples. The relationship between E. coli O86 and O127 was also determined by comparing the two O-antigen gene clusters. PMID- 15778031 TI - Genetic diversity of Mycobacterium avium subspecies paratuberculosis isolates from goats detected by pulsed-field gel electrophoresis. AB - Paratuberculosis in goats occurs worldwide causing considerable economic losses mainly due to reduced milk production. Nowadays, there is still relatively little knowledge about the epidemiology of this disease in goats, and only a few epidemiological studies have been carried out in goats naturally infected with Mycobacterium avium subspecies paratuberculosis (M. a. paratuberculosis). The objective of this study was to characterize forty four clinical caprine isolates of M. a. paratuberculosis by different molecular techniques (pulsed-field gel electrophoresis [PFGE], restriction fragment length polymorphism analysis coupled with hybridization to IS900, and IS1311 polymerase chain reaction-restriction enzyme analysis) to determine the most useful technique for molecular typing of caprine isolates, as well as to disclose the genetic variation amongst caprine isolates and the relationship with strains isolated from other animal species. PFGE was found to be the most discriminative technique identifying a total of 13 'multiplex' PFGE profiles, ten of which were novel profiles found only in caprine isolates to date. All isolates were genotyped as Type II strains, except two isolates that resembled the intermediate group referred as Type III. PMID- 15778032 TI - Sandwich ELISA detection of Clostridium perfringens cells and alpha-toxin from field cases of necrotic enteritis of poultry. AB - Sandwich ELISAs (sELISAs) for the detection of Clostridium perfringens cells and alpha-toxin were developed and used to screen intestinal samples from normal broiler chickens and from clinical cases of necrotic enteritis. The assays clearly distinguished between the two sets of samples. The sELISA absorbance values from samples obtained from the majority of healthy birds were low and those from the majority of necrotic enteritis cases were high. Together, the assays provide a suitable test for the rapid screening for the diagnosis of necrotic enteritis in poultry. PMID- 15778033 TI - Genetic diversity of Staphylococcus aureus isolated from ovine intramammary infections in Norway. AB - Three hundred and eighty-four Staphylococcus aureus isolates obtained from mammary secretions from 332 ewes kept for meat production were typed by pulsed field gel electrophoresis (PFGE). The ewes were from 242 flocks located in 13 counties distributed in four regions of Norway. In total, 64 different pulsotypes were identified, 31 of these were represented by a single isolate. Fifty-nine percent of the isolates belonged to one of five closely related pulsotypes. This group of pulsotypes occurred in all the counties. Although widely disseminated, the proportions of the prevalent and closely related pulsotypes differed between the regions. Nine pulsotypes were unique to single regions but the number of isolates belonging to each of these pulsotypes was low. Resistance to penicillin was found in only 3 of the 384 S. aureus isolates. These represented three different single banding patterns, not related to any of the prevalent pulsotypes found. PMID- 15778034 TI - Adhesion properties, fimbrial expression and PCR detection of adhesin-related genes of avian Escherichia coli strains. AB - Forty-nine avian pathogenic Escherichia coli (APEC) strains obtained from chickens suffering from septicemia (24), swollen head syndrome (14) and omphalitis (11), isolated from individuals in different regions of Brazil and from different outbreaks, were studied for their adhesion to trachea epithelial cells, fimbrial expression and hemagglutination capacity to different erythrocyte types. These results were compared with their content of fimbriae-related genes as detected by polymerase chain reaction (PCR) using specific pair of primers. The aim of these assays was to determine the importance of expression of adhesins in the pathogenic strains and to evaluate the presence of adhesin genes either previously described or not yet recognized for APEC strain. Thirty commensal strains isolated from poultry showing no signs of any of the above diseases were used to compare the results with the pathogenic isolates. The PCR assay demonstrated that septicaemic and swollen head syndrome strains had the highest number of adhesion-related genes of recognized importance in pathogenicity. Using different media for growth conditions, 40 different D-mannose resistant haemagglutination patterns were observed in this study, what indicates the expression of a great variability of surface agglutinins in these bacterial strains. Our results also showed that adhesion, whether D-mannose resistant (MRA) or D-mannose sensitive (MSA), is a characteristic observed in both pathogenic and commensal strains. Several strains with positive adherence had no genetic sequences related to the studied adhesin genes what indicates that our APEC strains probably possess a genome with adhesins genes besides those describe elsewhere and that have not yet been described. PMID- 15778035 TI - Detection of antibodies against Anaplasma marginale in milk using a recombinant MSP5 indirect ELISA. AB - An indirect enzyme linked immunosorbent assay (iELISA) for diagnosis of anaplasmosis using undiluted individual milk samples from dairy cows was developed. The recombinant 19 kDa major surface protein 5 (rMSP5) of Anaplasma marginale was used as antigen. A monoclonal antibody against bovine IgG1 conjugated with peroxidase and the chromogen 3,5,3',5'-tetramethylbenzidine were used in the test. Strong and weak, positive and negative milk samples were set up as reference controls. Results were expressed as percentage of positivity (PP) contrasting with the strongest positive control. The test was evaluated in two groups (G1 and G2) of lactating dairy cows from herds located in A. marginale non endemic areas of Argentina. The infection status of both groups, G1 (n=128) sampled after anaplasmosis outbreak, and G2 (n=216) free of anaplasmosis was established by polymerase chain reaction (PCR). Serum samples of cows from G1 and G2 were analyzed by card agglutination test (CAT) and competitive ELISA (cELISA), while the novel iELISA was evaluated in their corresponding milk samples. At a cutoff of 42 PP, the ELISA has 98% sensitivity and 95% specificity. A significant difference (P<0.0001) was found between the mean PP value of negative samples from G1 (17.4+/-14.9), and G2 (8.6+/-7.1). The agreement and kappa (kappa) value between iELISA and PCR was 96%, kappa=0.919; between iELISA and CAT was 97%, kappa=0.880; and between iELISA and cELISA was 97%, kappa=0.899. These results strongly support the usefulness of iELISA to detect A. marginale antibodies in milk. Additional studies are necessary to define the ability of the milk iELISA to detect not only acutely infected, but also carrier cattle. PMID- 15778036 TI - Prevalence of neutralizing antibodies to Equine rhinitis A and B virus in horses and man. AB - Equine rhinitis viruses (ERVs) are the causative agents of mild to severe upper respiratory infections in horses worldwide. Immunologically, four serotypes of ERVs have been identified. Equine rhinitis A virus (ERAV) and Equine rhinitis B virus 1 (ERBV1) are the most frequent serotypes in Europe. Both viruses have a broad host range in cultured cells with ERAV being able to infect humans. Since there is neither information on the seroprevalence of ERAV and ERBV1 in Austria nor on the zoonotic potential of ERBV1, we investigated 200 horse and 137 veterinary sera for the presence of neutralizing antibodies relating to ERAV and ERBV1. One hundred and eighty (90%) and 173 (86%) horse sera neutralized ERAV and ERBV1, respectively. In contrast, only four (2.7%) and five (3.6%) human sera showed weak neutralizing activity to ERAV and ERBV1, respectively. These results indicate that ERAV and ERBV1 are widespread in the Austrian horse population; however, the risk of acquiring zoonotic infection among veterinarians appears low. PMID- 15778038 TI - Heat treatment effects on Mycobacterium spp. isolated from ruminants in Nepal. PMID- 15778037 TI - Mutations in the quinolone-resistance determining region (QRDR) of Salmonella strains isolated from pigs in Spain. AB - Quinolone-resistance determining region (QRDR) of Gyrase A gene was sequenced in 54 Salmonella strains of pig origin that have different quinolone-resistance patterns. Those strains accounted for 12 different serotypes. Mutations at Ser83 or Asp87 were predominant in the studied isolates. However, for serotypes Anatum and Virchow, resistance to quinolones seemed to be linked to specific mutations, namely, Ser83-->Tyr and Ser83-->Phe, respectively. Other mutations found in different positions did not seem to have clinical significance except for changes at Asp82. PMID- 15778039 TI - Microcapsules prepared with different biomaterials to immobilize GDNF secreting 3T3 fibroblasts. AB - Cell microencapsulation represents a promising tool for the treatment of many central nervous system (CNS) diseases such as Parkinson's disease. In this technology, cells are surrounded by a semipermeable membrane which protects them from mechanical stress and isolates them from host's immune response. However, if the future clinical application of this strategy is wanted, many challenges remain including the improvement of the mechanical resistance of the microcapsules and the optimization of the intracapsular microenvironment conditions. In this way, the selection of the matrix is essential because the morphological and the physiological behavior of the cells depend on the interactions between the matrix and the enclosed cells. Assuming these considerations, three types of microcapsules elaborated with four different polymers: alginate, cellulose sulfate, agarose and pectin have been fabricated and compared in order to evaluate some key properties such as morphology, size and mechanical stability. Furthermore, GDNF secreting Fischer rat 3T3 fibroblasts were immobilized in each type of capsule and the viability and neurotrophic factor release was determinated. Results showed that the alginate and pectin microcapsules were the most resistant devices, maintaining an adequate microenvironment for the enclosed cells. In contrast, cells entrapped in alginate cellulose sulfate matrices presented the lowest mechanical resistance, cell viability and GDNF production. PMID- 15778040 TI - Evaluation of a water-soluble bioadhesive patch for photodynamic therapy of vulval lesions. AB - An innovative bioadhesive patch intended primarily as a vulval drug delivery system and, specifically, as a means to deliver photosensitisers, or their prodrugs, for photodynamic purposes is described. The patch was formulated with a copolymer of methyl vinyl ether and maleic anhydride (PMVE/MA) as a bioadhesive matrix and poly(vinyl chloride) as a drug-impervious backing layer. Adhesive strength to neonate porcine skin, as a model substrate, was evaluated using peel and tensile testing measurements. Acceptabilities of non-drug loaded patches were appraised using human volunteers and visual-analogue scoring devices. An optimal formulation, with water uptake and peel strengths appropriate for vulval drug delivery, was cast from a 20% (w/w) PMVE/MA solution and adhered with a strength of approximately 1.7 Ncm(-2). Patient evaluation demonstrated comfort and firm attachment for up to 4h in mobile patients. Aminolevulinic acid, a commonly used photosensitiser, was formulated into the candidate formulation and applied to vulval intraepithelial neoplastic lesions. Fluorescence under ultraviolet illumination revealed protoporphyrin synthesis. The patch achieves the extended application times obligatory in topical photodynamic therapy of vulval lesions, thereby contributing to potential methods for the eradication of neoplastic lesions in the lower female reproductive tract. PMID- 15778041 TI - Glycerin fatty acid esters as a new lubricant of tablets. AB - Lubrication properties were compared among glycerin fatty acid esters (Poem TR-FB and Poem TR-HB), magnesium stearate (Mg-St), and a sucrose fatty acid ester (RYOTO SUGAR ESTER S-370F: SSE). Granules containing 50% acetaminophen were prepared, and improvements in their fluidity by the lubricants were compared. The lubricant effects of TR-FB and HB during tablet punching (pressure transmission ratio, ejection force) were similar to those of Mg-St and were better than those of SSE. When the lubricant content, mixing time, and tabletting pressure were changed, TR-FB and TR-HB provided better tablet hardness than Mg-St. TR-FB and TR HB made tablets more disintegratable than Mg-St. When the effects of these lubricants on the stability of acetylsalicylic acid (ASA) were compared, Mg-St promoted its hydrolysis, but TR-FB or TR-HB did not affect its stability. PMID- 15778042 TI - Release characterization of dimenhydrinate from an eroding and swelling matrix: selection of appropriate dissolution apparatus. AB - The objective of this study was to evaluate the effect of various hydrodynamic conditions on drug release from an eroding and gel forming matrix. For this purpose, dimenhydrinate was formulated with hydroxypropyl methyl cellulose and polyethylene oxide into matrix tablets and the drug release in deionized water was evaluated spectrophotometrically, using multiple dissolution methods, namely, compendial USP 27-apparatus I-III, and a modified apparatus II (paddle over mesh). Various hydrodynamic conditions were examined at the agitation rates of 50 and 100 rpm for apparatus I and II, and 5 and 8 dpm for apparatus III. Similarity and difference factors were calculated using compendial apparatus II release data as reference. Among the methods, apparatus I showed the slowest initial release, while the release from apparatus III at 8 dpm was the highest among the methods. This was further compared via the dissolution half-times and calculation of the average release rate for each method. Based on the analysis of difference and similarity factors (f(1) and f(2)), the study clearly demonstrates the significance of hydrodynamics and the choice of a dissolution method and their respective effect on overall release profiles when erodible and swellable matrix systems are involved. Full surface exposure with insertion of mesh device in apparatus II may provide more realistic conditions especially when release data are to be used in developing IVIVCs. PMID- 15778043 TI - Usefulness of coadministration of bucolome in warfarin therapy: pharmacokinetic and pharmacodynamic analysis using outpatient prescriptions. AB - Bucolome, a nonsteroidal anti-inflammatory drug, has often been coadministered to patients who take warfarin as an anticoagulant. This combination increases the anticoagulant effect, which is most likely due to the interaction of bucolome with the pharmacokinetics (PK) or pharmacodynamics (PD) of warfarin. More than 30 years ago the mechanism of this interaction was reported to be inhibition of warfarin protein binding by bucolome, and the inhibition of warfarin metabolism by bucolome was also recently reported. Here, we examined daily doses of warfarin and its anticoagulant effect (thrombo-test, TT) in outpatient prescriptions in five hospitals to elucidate the drug interaction and the usefulness of this drug combination. Among the warfarin prescriptions, 78 were for patients also taking bucolome and 99 were for patients not taking bucolome. The daily dose of warfarin in patients taking bucolome was significantly lower than those without bucolome (ca. 40%). TT in patients taking bucolome was significantly lower as compared to those not taking bucolome. Control of the anticoagulant effect was greater with coadministration of bucolome and warfarin than with warfarin alone. PK and PD analysis of our results suggests that the improved therapeutic effect resulting from coadministration of warfarin with bucolome was due to lower and less patient to-patient variation of intrinsic hepatic clearance (CL(int)) of warfarin, since bucolome decreased the high CL(int) but did not have a great effect on the low CL(int). In conclusion, administration of bucolome in warfarin therapy is useful to control the anticoagulant effect of warfarin. Attention should also be paid to the enzymatic inhibition by bucolome on the PK of coadministered drugs. PMID- 15778044 TI - Molecular interaction in alginate beads reinforced with sodium starch glycolate or magnesium aluminum silicate, and their physical characteristics. AB - Diclofenac calcium-alginate (DCA) beads were reinforced with different amounts of sodium starch glycolate (SSG) or magnesium aluminum silicate (MAS) and were prepared using ionotropic gelation method. Complex formation of sodium alginate (SA) and SSG or MAS in calcium-alginate beads was revealed using FTIR spectroscopy. Differential scanning calorimetric study indicated that diclofenac sodium (DS) in amorphous form was dispersed in the matrix of DCA beads. The thermal behavior of SSG-DCA and MAS-DCA beads was similar to the control bead. Both additives can improve the entrapment efficiency of DCA beads. The swelling and water uptake of the beads depended on the properties of incorporated additives. The SSG-DCA beads showed a higher water uptake and swelling than MAS DCA beads. Moreover, the swelling of the beads showed a good correlation with the square root of time. The release kinetic of the beads in pH 6.8 phosphate buffer was swelling controlled mechanism, while that in distilled water followed Higuchi's model. The slower release rate and the longer lag time in pH 6.8 phosphate buffer was obtained from the SSG-DCA and MAS-DCA beads because of complex formation between SA and SSG or MAS. However, SSG in the beads could increase the release of DS from the beads in distilled water because it acted as a channeling agent. In contrast, MAS retarded the release of DS from the beads in distilled water due to the stronger matrix formation. PMID- 15778045 TI - Carrageenans can regulate the pulmonary absorption of antiasthmatic drugs and their retention in the rat lung tissues without any membrane damage. AB - Effects of various viscous vehicles on pulmonary absorption of antiasthmatic drugs were examined by an in situ pulmonary absorption experiment. Theophylline and fluticasone propionate were used as antiasthmatic drugs. The serum concentration-time profile of theophylline without viscous vehicles was similar to that following the intravenous injection, indicating that pulmonary absorption of theophylline was rapid and absolute. The serum concentration of theophylline was not controlled in the presence of 5% gelatin or 2% sodium alginate. However, 1% iota-carrageenan could control and regulate the serum concentration of theophylline. In the pharmacokinetic analysis, the C(max) values of theophylline significantly decreased, and its T(max) values increased in the presence of 1% and 2% iota-carrageenan, 1% kappa-carrageenan, and 2% sodium alginate compared with the control. The MRT and MAT values of theophylline with 1% iota-carrageenan were significantly higher than those without viscous vehicles. The local concentration of theophylline in the lung at 1h after intratracheal administration increased five-fold with 1% iota-carrageenan compared with the control. On the other hand, the pulmonary absorption of fluticasone propionate was controlled and regulated in the presence of 0.5% kappa-carrageenan. Additionally, the pulmonary inflammation after the exposure of carrageenans administered to the lung was evaluated in rats. Iota- and kappa-carrageenans did not cause local serious damage and inflammation to the pulmonary tissue. Therefore, these findings indicated that the carrageenans were effective to regulate the absorption rate of antiasthmatic drugs including theophylline and fluticasone propionate. PMID- 15778046 TI - Non-invasive vaccine delivery in transfersomes, niosomes and liposomes: a comparative study. AB - Non-invasive vaccine delivery is a top priority for public health agencies because conventional immunization practices are unsafe and associated with numerous limitations. Recently, the skin has emerged as a potential alternative route for non-invasive delivery of vaccine. Topical immunization (TI), introduction of antigen through topical application onto the intact skin, has many practical merits compared to injectable routes of administration. One of the possibilities for increasing the penetration of bioactives through the skin is the use of vesicular systems. Specially designed lipid vesicles are attracting intense attention and can be used for non-invasive antigen delivery. In the present study, elastic vesicle transfersomes, non-ionic surfactant vesicles (niosomes) and liposomes were used to study their relative potential in non invasive delivery of tetanus toxoid (TT). Transfersomes, niosomes and liposomes were prepared and characterized for shape, size and entrapment efficiency. These vesicles were extruded through polycarbonate filter (50-nm pore size) to assess the elasticity of the vesicles. The immune stimulating activity of transfersomes, niosomes and liposomes were studied by measuring the serum anti-TT IgG titre following topical immunization. The immune response elicited by topical immunization was compared with that elicited by same dose of alum-adsorbed tetanus toxoid (AATT) given intramuscularly. The results indicate that optimal formulations of transfersomes, niosomes and liposomes could entrap 72.7+/-3.4, 42.5+/-2.4 and 41.3+/-2.2% of antigen and their elasticity values were 124.4+/ 4.2, 29.3+/-2.4 and 21.7+/-1.9, respectively. In vivo study revealed that topically given TT containing transfersomes, after secondary immunization, could elicit immune response (anti-TT-IgG) that was equivalent to one that produced following intramuscularly alum-adsorbed TT-based immunization. In comparison to transfersomes, niosomes and liposomes elicited weaker immune response. Thus transfersomes hold promise for effective non-invasive topical delivery of antigen(s). PMID- 15778047 TI - Diethyl methyl chitosan as an intestinal paracellular enhancer: ex vivo and in vivo studies. AB - Chitosan exhibits favorable biological properties such as no toxicity, biocompatibility and biodegradability; therefore, it has attracted great attention in both pharmaceutical and biomedical fields. Chitosan exhibits poor solubility at pH values above 6 that prevents enhancing effects at the sites of absorption of drugs. In the present work, N-diethyl methyl chitosan (DEMC) was prepared and the enhancing effect of this polymer was investigated. Ex vivo studies have shown a significant increase in absorption of brilliant blue in the presence of diethyl methyl chitosan in comparison with chitosan. DEMC with positive charges is able to interact with tight junctions of colon epithelial cells and hence increases permeability of brilliant blue across the tight junctions. In vivo investigations have exhibited the absorption enhancer effects of DEMC on the colon absorption of insulin in normal and diabetic rats. The insulin absorption from the rat's colon was evaluated by its hypoglycemic effect. A significant decrease in blood glucose was observed, when mixture of insulin and DEMC was introduced in ascending colon of rats. PMID- 15778048 TI - Solubilisation in aqueous micellar solutions of block copoly(oxyalkylene)s. AB - The solubilisation capacities of micellar solutions of diblock and triblock copolymers composed of hydrophilic poly(ethylene oxide) and hydrophobic poly(styrene oxide) have been compared using the poorly water-soluble drug griseofulvin as a model solubilisate. Our results showed an increase of solubilisation capacity (expressed as mg griseofulvin per gram of hydrophobic block) with temperature and, for spherical micelles, with core volume before reaching limiting values. A change of micelle shape from spherical to cylindrical (or worm-like) resulting from an increase in micelle aggregation number was accompanied by a further enhancement of solubilisation capacity. Comparison with the solubilisation of the same drug in micellar solutions of block copolymers of poly(ethylene oxide) and poly(1,2-butylene oxide) showed that the solubilisation capacity of a poly(styrene oxide) block was approximately four times that of a poly(1,2-butylene oxide) block for spherical micelles. Solubilisation capacity at 25 degrees C was approximately doubled when griseofulvin was incorporated into a copolymer melt and micelles initially formed from the drug-loaded melt at 65 degrees C rather than by loading the drug into pre-micellised solution at 25 degrees C in the usual manner. PMID- 15778049 TI - Accelerated aging: prediction of chemical stability of pharmaceuticals. AB - Methods of rapidly and accurately assessing the chemical stability of pharmaceutical dosage forms are reviewed with respect to the major degradation mechanisms generally observed in pharmaceutical development. Methods are discussed, with the appropriate caveats, for accelerated aging of liquid and solid dosage forms, including small and large molecule active pharmaceutical ingredients. In particular, this review covers general thermal methods, as well as accelerated aging methods appropriate to oxidation, hydrolysis, reaction with reactive excipient impurities, photolysis and protein denaturation. PMID- 15778050 TI - Improvement of absorption enhancing effects of n-dodecyl-beta-D-maltopyranoside by its colon-specific delivery using chitosan capsules. AB - In general, absorption enhancing effects of various absorption enhancers were greater in the large intestine than those in the small intestinal regions. Therefore, the effectiveness of absorption enhancers is expected to be remarkably observed, if these enhancers can be delivered to the large intestine with some poorly absorbable drugs after oral administration. In this study, therefore, we examined whether chitosan capsules were effective for the colon-specific delivery of a certain absorption enhancer and can improve the absorption enhancing action of the absorption enhancer after oral administration. 5(6)-Carboxyfluorescein (CF) was used as a model drug to investigate the site-dependent effectiveness of various absorption enhancers by an in situ closed loop method. Sodium glycocholate (NaGC), n-dodecyl-beta-d-maltopyranoside (LM), sodium salicylate (NaSal) and sodium caprate (NaCap) were used as models of absorption enhancers in this study. Overall, the absorption enhancing effects of these enhancers for intestinal absorption of CF were greater in the colon than those in the jejunum and the ileum. Especially, among these enhancers tested in this study, LM showed much greater absorption enhancing effect in the colon than in the jejunum and the ileum. Therefore, LM was selected as a model absorption enhancer to examine the effect of chitosan capsules on the absorption enhancing effect of LM. When CF and LM were orally administered to rats using chitosan capsules, the plasma concentration of CF was much higher than those in other dosage forms including solution and gelatin capsules. Therefore, chitosan capsules may be useful carriers for colon-specific delivery of LM, thereby increasing its absorption enhancing effect from the intestinal membranes. PMID- 15778051 TI - Cremophor EL releases cyclosporin A adsorbed on blood cells and blood vessels, and increases apparent plasma concentration of cyclosporin A. AB - We examined the influence of cremophor EL (crEL) on the disposition kinetics of CyA in rats. A dose of 10mg/kg of CyA in a volume of 750 microL containing 4.3, 16 or 30% concentration of crEL was intravenously administered over 1 min to rats. The values of distribution volume at the steady-state (Vd(ss)) and total clearance (CL(tot)) of CyA in the presence of increasing amounts of crEL were decreased to about 1/3-1/5 of those with 4.3% crEL, in a crEL concentration dependent manner. The values of blood to plasma concentration ratio (RBP) and the apparent tissue to plasma concentration ratio (K(p,app)) of CyA with 30% crEL were both only about 1/2 of those of CyA with 4.3% crEL. Next, rats were intravenously given 30% crEL solution at 30 min after an intravenous administration of CyA (10 mg/kg) with 4.3% crEL. Subsequently, the blood and plasma concentrations of CyA rose significantly to 2.4 and 4.7 times those seen when i.v. 30% crEL was not given, respectively. In an in vitro study, we found that the uptake of CyA by red blood cells is inhibited by crEL, and that CyA adsorbed on the inner surface of blood vessels after the administration of CyA is released by crEL. The disposition kinetics of CyA is altered by i.v. administration in combination with the surfactant vehicle crEL, in a crEL concentration-dependent manner. PMID- 15778052 TI - Preparation, characterization, and tableting of a solid dispersion of indomethacin with crospovidone. AB - A significant problem with solid dispersion (SD) systems is the difficulty in preparing dosage forms. This difficulty can be overcome using crospovidone (CrosPVP) as a carrier. A powder SD of indomethacin (IM) with CrosPVP was prepared using mechanical mixing followed by heating to temperatures below the melting point. IM and CrosPVP interacted to produce IM in an amorphous state when its concentration was <40%. The solubility of IM was improved about fourfold compared to IM crystal. The SD had good fluidity, and tablets were prepared by direct compression. Tablets with small weight variation and acceptable hardness were obtained using only 1% of magnesium stearate as excipient. The dissolution of IM from tablets was similar to that of SD powder because CrosPVP, a disintegration agent, caused the tablets to break up rapidly. PMID- 15778053 TI - Solid dispersion particles of amorphous indomethacin with fine porous silica particles by using spray-drying method. AB - The solid dispersion particles of indomethacin (IMC) were prepared with different types of silica, non-porous (Aerosil 200) or porous silica (Sylysia 350) by using spray-drying method. Powder X-ray diffraction analysis showed that IMC in solid dispersion particles is in amorphous state irrespective of the type of silica formulated. In DSC analysis, the melting peak of IMC in solid dispersion particles with Sylysia 350 shifted to lower temperature than that in solid dispersion particles with Aerosil 200 although the peak of each solid dispersion particles was much smaller than that of original IMC crystals. Dissolution property of IMC was remarkably improved by formulating the silica particles to the solid dispersion particles. In comparing the effect of the type of the silica particles, the dissolution rate of solid dispersion particles with Sylysia 350 was faster than that with Aerosil 200. The formulation amount of IMC did not affect on the amorphous state of IMC in the resultant solid dispersion particles in powder X-ray diffraction patterns. However, the area of the melting peak of IMC in the solid dispersion particles increased and an exothermic peak owing to recrystallization was observed with increasing the IMC content in the DSC patterns. The dissolution rate of IMC from the solid dispersion particles with Sylysia 350 was faster than that of Aerosil 200 irrespective of IMC content. In stability test, amorphous IMC in the solid dispersion particles with each silica particles did not crystallize under storing at severe storage conditions (40 degrees C, 75% RH) for 2 months, while amorphous IMC without silica easily crystallized under same conditions. PMID- 15778054 TI - Preparation of molecularly imprinted cyclodextrin microspheres. AB - Molecularly imprinted cyclodextrins (MI-CDs) are prepared by cross-linking CDs in the presence of a template molecule. The binding ability of MI-CDs to the template molecule is specific; therefore, MI-CDs will prove to be useful materials. In this study, we prepared microspheres of MI-CDs (MSs-MI-CD) in a dimethylsulfoxide/poly(dimethylsiloxane) (PDMS) emulsion, using cholesterol as the template molecule. MSs-MI-CD were prepared under various conditions and were evaluated with respect to their morphology, size, and binding ability. MSs-MI-CD prepared at 65 degrees C were in an aggregated form; however, we could prepare separated and uniform MSs-MI-CD at 95 degrees C. The viscosity of PDMS influenced the size of MSs-MI-CD. The mean particle diameters of MSs-MI-CD prepared with 50 and 1000 mm(2)/s PDMS were 146 and 43 microm, respectively. The binding ability of MSs-MI-CD to cholesterol was higher than that of non-imprinted microspheres. Cholesterol imprinting also promoted the binding ability to other steroids; however, the increase in binding ability was most remarkable in the case of cholesterol, suggesting that we successfully introduced the cholesterol-specific binding ability into MSs-MI-CD. The novel MSs-MI-CD preparation method is useful and simple, and it will provide opportunities for further studies on the specific binding ability of MI-CDs. PMID- 15778055 TI - Measuring drug concentrations using pulsatile microdialysis: theory and method development in vitro. AB - A novel method of rapidly sampling drug concentrations, based on pulsatile microdialysis (PMD), was developed. In PMD, a dialysate fluid is pumped into a microdialysis probe, allowed to occupy the probe while at rest for some time, and then flushed at a high rate. A model that is based on a Fick's Laws was solved and tested, using methazolamide (MTZ) as the test drug in a variety of experimental setups, including time-dependent donor concentrations. Calibration plots of the donor versus sample concentrations were linear. There was excellent agreement between the calculated and experimental values of the fraction recovered obtained from the calibration plots. In a system for which the donor concentration declined in a first order manner, the data obtained using PMD and direct sampling of the donor were in nearly exact agreement with the theoretical value of k=0.09 min(-1). PMD was also able to collect data points quickly enough to characterize the rapid binding kinetics of MTZ by activated charcoal. It was concluded that PMD is an accurate method of sampling drug concentrations, and can obtain samples over shorter time intervals and more frequently than previously available methods. PMID- 15778056 TI - Prediction of oral drug absorption in humans by theoretical passive absorption model. AB - The purpose of the present study was to examine the oral drug absorption predictability of the theoretical passive absorption model (TPAM). As chemical descriptors of drugs, the octanol/buffer distribution coefficient at pH 6.0 (D(ow)), intrinsic octanol-water partition coefficient (P(ow)), pK(a), and molecular weight (MW) were calculated from the chemical structure. Total passive intestinal membrane permeation consists of transcellular, paracellular and unstirred water layer (UWL) permeation. Transcellular permeation was modeled based on the pH-partition hypothesis with correction for cationic species permeation, and the independent variables were D(ow), P(ow), and pK(a). Paracellular permeation was modeled as a size-restricted diffusion within a negative electrostatic field-of-force, and the independent variables were MW and pK(a). UWL permeation was modeled as diffusion across a water layer, and the independent variable was MW. Cationic species permeation in the transcellular permeation model and the effect of a negative electric field-of-force in the paracellular permeation model were the extensions to the previous TPAM. The coefficients of the paracellular and UWL permeation models were taken from the literature. A data set of 258 compounds with observed values of Fa% (the fraction of a dose absorbed in humans) taken from the literature was employed to optimize four fitting coefficients in the transcellular permeation model. The TPAM predicted Fa%, with root mean square errors of 15-21% and a correlation coefficient (CC) of 0.78-0.88. In addition, the TPAM predicted the effective human intestinal membrane permeability with a CC of 0.67-0.77, as well as the contribution of paracellular permeation. The TPAM was found to predict oral absorption from the chemical structure of drugs with adequate predictability for usage in drug discovery. PMID- 15778057 TI - Ketorolac amide prodrugs for transdermal delivery: stability and in vitro rat skin permeation studies. AB - Various amide prodrugs of ketorolac were synthesized and their rat skin permeation characteristics were determined. The solubility of the prodrugs in propylene glycol (PG) was determined at 37 degrees C while lipophilicity was obtained as 1-octanol/water partition coefficient (logP) and capacity factor (k') using HPLC. Stability of the prodrugs in rat skin homogenate, plasma and liver homogenate was investigated to observe the enzymatic degradation. Rat skin permeation characteristics of the prodrugs saturated in PG were investigated using the Keshary-Chien permeation system at 37 degrees C. The logP value of the prodrugs increased up to 4.28 with the addition of various alkyl chain to ketorolac which has a logP of 1.04. Good linear relationship between logP and capacity factor was observed (r(2)=0.89). Amide prodrugs were converted to ketorolac only in rat liver homogenate. However, the skin permeation rate of amide prodrugs did not significantly increase, probably due to their low aqueous solubility. Chemical modification of the ketorolac amide prodrug and/or the selection of proper vehicle to increase aqueous solubility would be necessary for an effective transdermal delivery of ketorolac. PMID- 15778058 TI - In vivo evaluation of tablets and capsules containing spray-dried o/w-emulsions for oral delivery of poorly soluble drugs. AB - It is recognised that poorly soluble drugs may show an increased oral bioavailability when incorporated in o/w-emulsions. Encapsulating the emulsion lipid droplets in hydroxypropyl methylcellulose (HPMC) by spray drying has been demonstrated to preserve an improved bioavailability releasing lipid droplets from the powder in vivo. However, the spray-dried powder is cohesive and bulky requiring additional processing to improve handling. This was resolved in previous work where a directly compressible dry emulsion formulation was described. The purpose of the present study is to investigate the oral bioavailability resulting from administration of a directly compressible dry emulsion as a tablet and compare it with a HPMC dry emulsion powder and a simple lipid solution. Four female Beagle dogs received a single dose of each formulation containing the same amount of medium-chain triglycerides (MCT) and a model drug, Lu 28-179. Cyclodextrin solutions administered orally and intravenously were used as references. The absolute bioavailability decreased in the order cyclodextrin solution (0.14), HPMC dry emulsion (0.11), technically improved dry emulsion (0.10) and MCT solution (0.06). The directly compressible dry emulsion tablets were concluded to be comparable to a HPMC dry emulsion powder in terms of bioavailability. The lack of statistically significant differences relative to a MCT solution was ascribed to a low and variable absolute oral bioavailability of the model drug. PMID- 15778059 TI - Transdermal delivery of naloxone: skin permeation, pharmacokinetic, irritancy and stability studies. AB - The current investigation aims to evaluate ex vivo, in vivo performance, stability and irritancy potential of a transdermal formulation of naloxone (NLX) developed at our laboratory at different concentrations (10, 20 and 30mg/g of gel) in a transdermal reservoir patch. Ex vivo permeation studies were performed by employing porcine and rat skins. In vivo performance was assessed in Sprague Dawley rats by single and multiple application of the patch. Further stability of the formulation was established for 3 months at accelerated stability conditions as per ICH guidelines. Amongst the barriers used the rat skin was found to be more permeable than the porcine epidermis and the flux across each barrier increased with increasing thermodynamic activity of drug in the gel. Based on ex vivo data, the surface area (SA) of the patch was predicted to be 39.6 cm(2) in order to achieve therapeutic blood levels. Upon single dose administration, the steady-state levels were maintained from 4-48 h, which proves the clear advantage of transdermal delivery system over the current mode of administration, i.e., intravenous (i.v.) bolus which is effective upto a maximum of 1.5h. Upon multiple dose administration, the sustained steady state for 12h, even after patch removal proves the formation of drug depot in the skin. The formulations were found to be stable with respect to NLX assay and penetration enhancer efficacy upto 3 months under accelerated stability conditions. The alteration of penetration barrier function, as evidenced by increased trans epidermal water loss (TEWL) was not accompanied by any significant amount of skin irritation measured using laser doppler velocimetry (LDV). The developed transdermal delivery system of NLX is efficacious, stable and safe upon single and multiple dose applications each lasting for 48 h. PMID- 15778060 TI - Characterization of nifedipine microparticles prepared by hot air coating technique. AB - In the present work, the Hot Air Coating (HAC) technique was used to prepare microparticulate systems containing nifedipine. Binary mixtures constituting of nifedipine and cetearyl alcohol (CA) in different proportions (30:70, 50:50, 70:30) were studied: they were homogenized by mixing or milling before spray treatment and successively subjected to a coating procedure with the HAC apparatus fed with air at 120 degrees C under a pressure of 4.5 atm. Morphology, entrapment efficiency, drug stability, thermal behaviour and the drug dissolution profile of HAC-treated and non-treated materials were examined and compared. The HAC products show the possession of physical and physico-chemical properties and dissolution behaviour different from those of the initial physical mixtures. The operative conditions employed in the spray process allow the obtaining of microparticles containing relevant percentages of the drug (at least up to 50%). Moreover, the experimental results give evidence that the milling pre-treatment of mixtures, unlike mixing, has significant effects on the properties of the lipid-coated microparticles. PMID- 15778061 TI - Enhancement of the release of azelaic acid through the synthetic membranes by inclusion complex formation with hydroxypropyl-beta-cyclodextrin. AB - The aim of this study was to investigate the release rates of azelaic acid and azelaic acid-hydroxypropyl-beta-cyclodextrin (HPbetaCD) inclusion complex through three types of synthetic membranes, namely cellophane, silicone and elastomer membranes. Solid inclusion complexes of azelaic acid-HPbetaCD at the molar ratio of 1:1 were prepared by coevaporation and freeze-drying methods, subsequently characterized by differential scanning calorimetry, X-ray diffractometry and dissolution studies. Solid inclusion complex obtained by coevaporation method which exhibited the inclusion of azelaic acid in the HPbetaCD cavity and gave the highest dissolution rate of azelaic acid was selected for the release study. Release studies of azelaic acid and this complex through the synthetic membranes were conducted using vertical Franz diffusion cells at 30 degrees C for 6 days. The release rates of azelaic acid through the synthetic membranes were enhanced by the formation of inclusion complex with HPbetaCD at the molar ratio of 1:1, with the increasing fluxes of about 41, 81 and 28 times of the uncomplexed system in cellophane, silicone and elastomer membranes, respectively. The result from this study can be applied for the development of azelaic acid for topical use. PMID- 15778062 TI - In vitro drug release mechanism and drug loading studies of cubic phase gels. AB - Glyceryl monooleate/water cubic phase systems were investigated as drug delivery systems, using salicylic acid as a model drug. The liquid crystalline phases formed by the glyceryl monooleate (GMO)/water systems were characterized by polarizing microscopy. In vitro drug release studies were performed and the influences of initial water content, swelling and drug loading on the drug release properties were evaluated. Water uptake followed second-order swelling kinetics. In vitro release profiles showed Fickian diffusion control and were independent on the initial water content and drug loading, suggesting GMO cubic phase gels suitability for use as drug delivery system. PMID- 15778063 TI - Accelerated photostability study of tretinoin and isotretinoin in liposome formulations. AB - The photodegradation of retinoic acids, tretinoin and isotretinoin, in ethanol and liposomes was studied. The light irradiation was performed according to the conditions suggested by the ICH Guideline for photostability testing by using a Xenon lamp within a wavelength range of 300-800 nm. The photodegradation process was monitored by UV spectrophotometry. In ethanol solution, tretinoin and isotretinoin undergo complete isomerization just within a few seconds of light exposure to give 13-cis and 9-cis isomers, respectively. The 13-cis isomer from tretinoin undergoes in turn a slow isomerization to the same 9-cis isomer. Both retinoic acids incorporated in liposome complexes showed an increased stability in comparison to the ethanol solutions. In particular for tretinoin, a residual concentration of 60% was still present after a light irradiance of 3470 kJ/m(2), by means of a 250 W/m(2) light power for 240 min, versus a residual value of just 8% measured at the same time in ethanol solution. Moreover, the isomerization rate in liposomes resulted reduced for isotretinoin and practically irrelevant for tretinoin. The degradation rate was found to be dependent on the drug concentration. The better stability of the tretinoin in liposome complex was supposed to be related to its higher incorporation value due to the linear structure of the molecule. PMID- 15778064 TI - Poly(ethylene oxide)-modified poly(epsilon-caprolactone) nanoparticles for targeted delivery of tamoxifen in breast cancer. AB - This study was carried out to evaluate and compare the biodistribution profile of tamoxifen when administered intravenously (i.v.) as a simple solution or when encapsulated in polymeric nanoparticulate formulations, with or without surface stabilizing agents. Tamoxifen-loaded, poly(ethylene oxide)-modified poly(epsilon caprolactone) (PEO-PCL) nanoparticles were prepared by solvent displacement process that allowed in situ surface modification via physical adsorption of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock polymeric stabilizer (Pluronic). The nanoparticles were characterized for particle size and surface charge. Presence of PEO chains on nanoparticle surface was ascertained by electron spectroscopy for chemical analysis (ESCA). In vivo biodistribution studies were carried out in Nu/Nu athymic mice bearing a human breast carcinoma xenograft, MDA-MB-231 using tritiated [(3)H]-tamoxifen as radio-marker for quantification. PEO-PCL nanoparticles with an average diameter of 150-250 nm, having a smooth spherical shape, and a positive surface charge were obtained with the formulation procedure. About 90% drug encapsulation efficiency was achieved when tamoxifen was loaded at 10% by weight of the polymer. Aqueous wettability, suspendability, and ESCA results showed surface hydrophilization of the PCL nanoparticles by the Pluronics. The primary site of accumulation for the drug-loaded nanoparticles after i.v. administration was the liver, though up to 26% of the total activity could be recovered in tumor at 6h post-injection for PEO-modified nanoparticles. PEO-PCL nanoparticles exhibited significantly increased level of accumulation of the drug within tumor with time as well as extended their presence in the systemic circulation than the controls (unmodified nanoparticles or the solution form). Pluronic surfactants (F-68 and F 108) presented simple means for efficient surface modification and stabilization of PCL nanoparticles to achieve preferential tumor-targeting and a circulating drug reservoir for tamoxifen. PMID- 15778065 TI - Pharmacological activity of peroral chitosan-insulin nanoparticles in diabetic rats. AB - The objective of the present study was to evaluate the effects of formulation parameters on the in vivo pharmacological activity of the chitosan-insulin nanoparticles. Chitosan-insulin nanoparticles were prepared by ionotropic gelation at pH 5.3 and 6.1 and denoted as F5.3 np and F6.1 np, respectively. F5.3 np and F6.1 np administered orally at insulin doses of 50 U/kg and/or 100 U/kg were effective at lowering the serum glucose level of streptozotocin-induced diabetic rats. The 100 U/kg-dose F5.3 np sustained the serum glucose at pre diabetic levels for at least 11 h. In comparison, F6.1 np had a faster onset of action (2h versus 10h) but lower efficiency. The effectiveness of peroral F5.3 np and F6.1 np in lowering the serum glucose level of streptozotocin-induced diabetic rats was ascribed to the local effect of insulin in intestine. Confocal micrographs showed strong interaction between rat intestinal epithelium and chitosan nanoparticles 3h post-oral administration. PMID- 15778066 TI - Pharmacokinetics and biodistribution of RGD-targeted doxorubicin-loaded nanoparticles in tumor-bearing mice. AB - We report the biodistribution and pharmacokinetics (PK) of a cyclic RGD doxorubicin-nanoparticle (NP) formulation in tumor-bearing mice. The NP core was composed of inulin multi-methacrylate with a targeting peptide, cyclic RGD, covalently attached to the NPs via PEG-400. Seventy-two percent of the doxorubicin was attached to the NP matrix via an amide bond; 28% of doxorubicin was entrapped as unconjugated drug. The PK of total, unconjugated and metabolized doxorubicin was examined for 5 days following intravenous (i.v.) administration of the NP formulation (250 microg doxorubicin equiv.), revealing a bi-exponential fix with a terminal half-life of 5.99 h. In addition, the biodistribution studies revealed decreasing drug concentrations over time in the heart, lung, kidney and plasma and accumulating drug concentrations in the liver, spleen and tumor. The drug concentration in these latter tissues peaked between 24 and 48 h with the liver, spleen and tumor containing 56, 3.5 and 1.8% of the administered dose at t=48 h, respectively. In contrast to all of the organs studied, the tumors contained high levels of a doxorubicin metabolite. PMID- 15778067 TI - Manipulation of local disposition and gene expression characteristics of plasmid DNA following intramuscular administration by complexation with cationic macromolecule. AB - To modulate the immune responses of DNA vaccine, it is very important to control the disposition and gene expression of plasmid DNA (pDNA) after local administration. We chose methylated bovine serum albumin (mBSA), a cationic macromolecule, as a carrier of pDNA. We examined the effects of complexation of pDNA with mBSA on the disposition and gene expression in mice after intramuscular administration. The elimination from injection site was retarded and the accumulation to lymph nodes was increased at the positively charged mBSA/pDNA complexes. As the charge ratios of mBSA/pDNA complexes were higher, the levels of gene expression were reduced. Antigen specific immune responses were evaluated using pDNA encoding ovalbumin (OVA), pCMV-OVA, as a model antigen-expressing pDNA. However, significant levels of production of anti-ovalbumin IgG antibody were obtained in mice immunized with a positively charged complex, mBSA/pCMV-OVA (8:1) (weight ratio). In vitro experiments using DC2.4 cells, a murine dendritic cell line, demonstrated that the levels of gene expression and cytokine release were increased by complexation. These results suggest that the immune responses might be manipulated by complexation presumably due to the altered disposition and gene expression of pDNA. PMID- 15778068 TI - Investigation of the release behavior of DEHP from infusion sets by paclitaxel loaded polymeric micelles. AB - The current clinical formulation of paclitaxel (Taxol) contains 1:1 blend of Cremophor EL (polyethoxylated castor oil) and dehydrated ethanol. Cremophor EL and dehydrated ethanol are well known to leach di-(2-ethylhexyl) phthalate (DEHP) from polyvinyl chloride (PVC) infusion bags and PVC administration sets. DEHP is a possible hepatotoxin, carcinogen, teratogen and mutagen. Long-term exposure to DEHP may cause health risks. As an alternative formulation for paclitaxel, paclitaxel-loaded polymeric micelles (PLPM), made of monomethoxy poly(ethylene glycol)-block-poly(d,l-lactide) (mPEG-PDLLA) diblock copolymer, has demonstrated clear advantages over Taxol in pharmacokinetics and therapeutic index. Paclitaxel in either PLPM or Taxol formulations, diluted in 0.9% sodium chloride injection, was stable in the PVC infusion bags. The PLPM formulation significantly reduced the amount of DEHP extracted from PVC infusion bags and PVC administration sets. For PLPM diluted in 0.9% sodium chloride injection, the total amount of DEHP delivered over the simulated infusion period was 0.7 mg for 3h and 2.0 mg for 24 h, which was less than 2.9% of the DEHP extracted by Taxol. These results confirmed that there is negligible risk of DEHP exposure from diluted PLPM i.v. infusion using PVC infusion bags and PVC administration sets. PMID- 15778070 TI - Reporting unplanned extubation. AB - Between 1995 and 2002 seven clinical audits were undertaken in consecutive periods over twelve months to determine the frequency and risk factors associated with reported unplanned extubation (UE) within a 22-bed general and surgical Intensive Care Unit (ICU). Nursing and medical staff provided information on the patient's age, diagnosis, mental status, precipitating causes and investigations/treatment ordered. Following the first audit, modifications were made to include anonymous reporting. Additional information was also obtained on the patient's position, sedation regimen, method of endotracheal tube (ETT) placement and the use of physical restraints. A clinical indicator was established to monitor the UE incidence based as a rate of UE per 100 patients. Audit results were between 1.06% and 4.86% with an aggregate rate from 1995 to 2002 of 2.6%. This rate compares favourably with the range of 2.8-22.5% reported in the literature. Over the survey periods, 28-60% of patients were assessed as being confused or agitated, 47-67% restrained and 53-70% sedated. The UE reported rate initially increased when anonymous reporting was introduced from 1.06% to 4.86%. Unplanned extubation incidence subsequently decreased in Surgical ICU following the introduction of clinical pathways, early weaning and nurse led extubation. Monitoring UE in ICU provides important information on the quality of care. We would recommend a system of anonymous reporting to more freely reflect incidence. PMID- 15778071 TI - A randomised controlled trial of weaning from mechanical ventilation in paediatric intensive care (PIC). Methodological and practical issues. AB - INTRODUCTION: Most children admitted to the Paediatric Intensive Care Unit (PICU) require assistance with breathing via a mechanical ventilator. Weaning from mechanical ventilation is the transition from ventilatory support to spontaneous breathing. Traditionally weaning has been with the authority of the medical staff. However, current opinion suggests that weaning could be performed by nurses using a standardised protocol [Schultz TR, Lin RJ, Watzman HM, Durning SM, Hales R, Woodson A, et al. Weaning children from mechanical ventilation: A prospective randomised trial of protocol-directed versus physician-directed weaning. Respir Care 2001;46(8):772-82]. The potential advantages of nurse-led (protocol-directed) weaning include: A reduction in weaning time and PICU stay with cost savings. Reduced complications. Improved quality of care. Appropriate use of resources. METHODS: A Randomised Controlled Trial was performed to test the null hypothesis: there is no difference between the clinical effectiveness of nurse-led versus medical-led weaning of infants from mechanical ventilation. Data was collected for 7 infants and analysed. RESULTS: Results indicated no significant differences between the two study groups. Unfortunately due to recruitment problems few inferences can be drawn from the data. CONCLUSION: The trial was unsuccessful due to Recruitment issues. Physical constraints. Impractical entry criteria. Limited randomisation service. Ethical constraints. Barriers to parental participation. The methods, the difficulties encountered and the implications for future research are addressed. PMID- 15778072 TI - The clinical and economic benefits of administering piperacillin-tazobactam by continuous infusion. AB - Beta-lactam antibiotics, such as piperacillin-tazobactam, are commonly administered frequently throughout the day as intermittent infusions over a period of 30-60 min. However, increasing knowledge of how these antibiotics kill bacteria has made continuous infusion a valuable option to achieve maximal clinical outcomes while consuming the least amount of institutional resources. Continuous infusion of piperacillin-tazobactam is currently used at our hospital because of its clinical and economic benefits when compared with intermittent infusion. This article will review our experience with the administration of piperacillin-tazobactam by continuous infusion and the numerous advantages we have documented. Additionally, advantages related to a reduction in nursing resource consumption will be a focus of discussion. PMID- 15778073 TI - Role of environmental cleaning in controlling an outbreak of Acinetobacter baumannii on a neurosurgical intensive care unit. AB - An outbreak of Acinetobacter baumannii colonization and infection occurred in 19 patients over a 14-month period during 1998-1999 on a neurosurgical intensive care unit. During efforts to control the outbreak a significant correlation was observed between the number of environmental isolates of A. baumannii obtained during each monthly screening and the number of patients with A. baumannii colonization/infection in the same calendar month (P=0.004). Use of 1000 ppm hypochlorite solution and the introduction of new cleaning protocols reduced the number of environmental isolates. Failure to maintain low levels of environmental contamination with A. baumannii resulted in increases in patient colonization. This study showed that high standards of cleaning play an integral role in controlling outbreaks of A. baumannii in the intensive care unit setting. PMID- 15778074 TI - Infection control issues in central venous catheter care. AB - Central venous catheters (CVCs) are now a routine part of patient management in the intensive care unit (ICU). Over time, a vast amount of literature associated with the use and care of CVCs has accumulated. The purpose of this article is to discuss the literature associated with the care of these devices in a narrative format. Although particular attention is paid to infection control issues, other fundamental areas such as catheter design, dressings, line changing and post insertion management are also discussed. The article goes on to look at the future of CVC design and concludes with an analysis of future developments related to CVCs. PMID- 15778075 TI - Finding the voice of clinical experience: participatory action research with registered nurses in developing a child critical care nursing curriculum. AB - The voice of clinical nurses is important to find and hear in the design of curricula. A participative action research project proposed to add this voice to the design of a new Critical Care Child Nursing programme at the University of Cape Town (UCT). Nurses' experiences of nursing critically ill children and their perceived learning needs in this context, were the central focus of the study. Participants were registered nurses working in the paediatric intensive care unit at the Red Cross Children's Hospital (a specialist hospital), which offers secondary and tertiary care in the Cape Town region and beyond. Data were gathered in five focussed group discussions. Findings indicate that the Critical Care Child Nurse needs not only a specialised knowledge base and acutely developed assessment skills, but also astute interpersonal skills. The nurse's professional identity and integration into the multidisciplinary team need exploring. Together with the development of interpersonal skills, the nurse needs to engage the child and family. PMID- 15778076 TI - Development of a new eye care guideline for critically ill patients. AB - Ventilated, critically ill patients have an increased risk of corneal exposure and microbial keratitis; however there is evidence that eye care is a neglected area of patient care. An audit of eye surface disease and eye care documentation revealed a problem within one general intensive care unit (GICU). An education programme was developed to highlight the incidence of eye surface disease and importance of eye care in the ventilated patient. PMID- 15778077 TI - Clinical research 4: qualitative data collection and analysis. AB - This six-part research series is aimed at clinicians who wish to develop research skills, or who have a particular clinical problem that they think could be addressed through research. The series aims to provide insight into the decisions that researchers make in the course of their work, and to also provide a foundation for decisions that nurses may make in applying the findings of a study to practice in their own Unit or Department. The series emphasises the practical issues encountered when undertaking research in critical care settings; readers are encouraged to source research methodology textbooks for more detailed guidance on specific aspects of the research process. A couple of points: 1. It is artificial to describe research as qualitative or quantitative. Studies often include both dimensions (for example, Evangelista LS, Doering L, Dracup K. Meaning and life purpose: the perspectives of post-transplant women. Heart Lung 2003;32(4):250-7; Fitzsimmons D, Parahoo K, Richardson SG, Stringer M. Patient anxiety while on a waiting list for coronary artery bypass surgery: a qualitative and quantitative analysis. Heart Lung 2003;32(1):23-31). However, for the purposes of this paper/series, this distinction is drawn for clarity of writing. 2. It is common practice for quantitative studies to refer to study 'subjects' and qualitative studies to refer to study 'participants'. For ease of reading, the latter term will be used throughout this series. PMID- 15778079 TI - Renal tubular acidosis: developments in our understanding of the molecular basis. AB - Renal tubular acidosis is a metabolic acidosis due to impaired acid excretion by the kidney. Hyperchloraemic acidosis with a normal anion gap and normal (or near normal) glomerular filtration rate, and in the absence of diarrhoea, defines this disorder. However, systemic acidosis is not always evident and renal tubular acidosis can present with hypokalaemia, medullary nephrocalcinosis and recurrent calcium phosphate stone disease, as well as growth retardation and rickets in children, or short stature and osteomalacia in adults. Renal dysfunction in renal tubular acidosis is not always confined to acid excretion and can be part of a more generalised renal tubule defect, as in the renal Fanconi syndrome. Isolated renal tubular acidosis is more usually acquired, due to drugs, autoimmune disease, post-obstructive uropathy or any cause of medullary nephrocalcinosis. Less commonly, it is inherited and may be associated with deafness, osteopetrosis or ocular abnormalities. The clinical classification of renal tubular acidosis has been correlated with our current physiological model of how the nephron excretes acid, and this has facilitated genetic studies that have identified mutations in several genes encoding acid and base ion transporters. In vitro functional studies of these mutant proteins in cell expression systems have helped to elucidate the molecular mechanisms underlying renal tubular acidosis, which ultimately may lead to new therapeutic options in what is still treatment only by giving an oral alkali. PMID- 15778078 TI - Astrocytes: regulation of brain homeostasis via apolipoprotein E. AB - Astrocytes are derived from the ventricular and subventricular zones of the neural plate, though there is controversy over their derivation from astrocyte specific precursor cells or radial glia intermediates. Astrocytes are the most abundant cell type in the brain and contribute to brain homeostasis in several ways, including buffering of extracellular K+, regulating neurotransmitter release, forming the blood-brain barrier (BBB), releasing growth factors, and regulating the brain immune response. In addition, astrocytes have been shown to release apolipoprotein E (ApoE), which has been shown to regulate neurotransmission, growth factor release, and immune responses. Due to the diverse functions of astrocytes, they may play a role in a variety of diseases such as hepatic encephalopathy, multiple sclerosis, epilepsy, and age-related diseases including Alzheimer's disease and Parkinson's disease. This review highlights many of the diverse roles played by astrocytes in regulating brain homeostasis and discusses their potential role in a variety of disorders. PMID- 15778080 TI - Platelet factor 4: a chemokine enigma. AB - Platelet factor 4 (PF4) is a platelet alpha-granule protein sequenced over 25 years ago that is a founding member of the C-X-C chemokine family, yet its physiologic function has yet to be definitively established. Initial investigations focused on possible procoagulant roles for PF4 in platelet function and plasmatic coagulation. Subsequent in vitro studies have, however, described a puzzling array of other apparently unrelated biologic functions, including inhibition of angiogenesis and hematopoiesis, promotion of neutrophil adhesion, and activation, enhancement of oxy-LDL binding to the LDL receptor and stimulation of anti-coagulant activated protein C generation by the thrombomodulin/protein C system. Preliminary studies with a just-described PF4 knockout mouse line support a role for PF4 in platelet-dependent thrombosis in vivo. PMID- 15778081 TI - FLT-3: a new focus in the understanding of acute leukemia. AB - The FMS-like tyrosine kinase-3 (FLT-3), which belongs to the class III receptor tyrosine kinase family, is primarily expressed by hematopoietic cells and plays an important role in hematopoiesis. FLT-3 is also expressed in the majority of acute leukemias, in which the presence of FLT-3 activating mutations is associated with poor prognosis. Consequently, there has been a recent surge in the development of FLT-3 inhibitors for the molecular targeting of leukemia, and many of these are now in clinical trials. An improved understanding of how FLT-3 interacts with its ligand, as well as how FLT-3 activating mutations are able to trigger downstream intracellular signaling pathways, will provide greater insight to how small molecule inhibitors may best be utilized and combined with established chemotherapeutic drugs for the management of patients with high-risk acute leukemia. PMID- 15778082 TI - Hephaestin--a ferroxidase of cellular iron export. AB - Hephaestin is a transmembrane copper-dependent ferroxidase necessary for effective iron transport from intestinal enterocytes into the circulation. Hephaestin is mutated in sex-linked anemia (sla) mice. The initial uptake of iron from the diet in these animals is normal, but the basolateral export of iron from enterocytes is defective, resulting in iron deficiency and microcytic hypochromic anemia. In addition to the small intestine, hephaestin is expressed to a lesser extent in colon, spleen, placenta and kidney but its role in these tissues remains unknown. So far, hephaestin has not been linked to a human disease. PMID- 15778083 TI - Opposite effects of peroxidase in the initial stages of tyrosinase-catalysed melanin biosynthesis. AB - The tyrosinase/oxygen enzymatic system catalyses the orthohydroxylation of L tyrosine to L-dopa and the oxidation of this to dopaquinone, which evolves non enzymatically towards to form melanins. The literature has demonstrated and revised the existence of peroxidase/hydrogen peroxide in the melanosomas of skin melanocytes, but points to controversy concerning the effects on melanogenesis. Some authors have recently proposed a new physiological function for tyrosinase, namely the direct scavenging of tyrosyl radicals, which are toxic oxidants of melanocytes. In this contribution, we describe and interpret four effects of peroxidase/hydrogen peroxide on melanogenesis. Two of these effects are its antagonism and synergy as regards the monophenolase and diphenolase activities, respectively, of tyrosinase/oxygen in the initial steps that trigger melanogenesis. Another effect concerns the increase in the oxidant character of the medium in the melanosome by increasing the synthesis of oxidising quinones (o dopaquinone, p-topaquinone, dopachrome) and the consumption of antioxidant diphenols (L-dopa), which are intermediate biomolecules in melanogenesis. Lastly, we demonstrate that the tyrosyl radicals generated by light or by the peroxidase/hydrogen peroxide system are not directly trapped by the tyrosinase but by the antioxidant orthodiphenol, L-dopa, accumulated in the steady-state of melanogenesis. In conclusion, peroxidase/hydrogen peroxide may help regulate the development of melanogenesis and the oxidant environment within the melanosome. This enzyme deserves further study for its possible antitumoral and depigmentation capacities in skin cancer and hyperpigmentation. PMID- 15778084 TI - Proteomic analysis of interleukin 6-induced differentiation in mouse myeloid leukemia cells. AB - Cytokine-induced differentiation of myeloid leukemia cells has important therapeutic implications, but the mechanism remains to be clarified. M1 cell, a mouse acute myeloid leukemia cell line, which underwent growth inhibition, terminal differentiation and apoptosis in response to IL-6, was selected as an experimental model to study on the molecular mechanisms of myeloid cell differentiation on a proteome-wide scale. Cell differentiation was evaluated by cell morphology and CD11b expression. With two-dimensional (2D) gel analyses, 17 protein spots showed obvious changes in quantity during the process of differentiation were found. With matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) or/and nano-electrospray ionization MS/MS (ESI-MS/MS) analysis, 15 protein spots were identified. The mRNA levels of these 15 proteins during differentiation were also examined using a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis. Except two proteins, the mRNA levels demonstrated similar expression patterns to what the proteomic analysis revealed. The identified proteins were known to be involved in different cellular functions, including protein synthesis, transcription, signal transduction, cell cycle control, cell rescue and defense, cellular organization, and metabolism. Notably, seven proteins were not described before to be involved in differentiation. Our data provide novel information for a better understanding of the mechanisms by which terminal differentiation of acute myeloid leukemia cells induced by IL-6. PMID- 15778086 TI - Antitumor effects of human ribonuclease inhibitor gene transfected on B16 melanoma cells. AB - Human ribonuclease inhibitor (RI) is a cytoplasmic acidic protein. The experiment demonstrated that it might effectively inhibit tumor-induced angiogenesis and inhibit tumor growth. Ribonuclease inhibitor is constructed almost entirely of leucine-rich repeats, which might be involved in unknown biological effects besides inhibiting RNase A and angiogenin activities. The exact molecular mechanism of antitumor on ribonuclease inhibitor remains unclear so far. In order to further understand the function of ribonuclease inhibitor and investigate the relationship with tumor growth, our study established a transfection of human ribonuclease inhibitor cDNA into the murine B16 cells by the retroviral packaging cell line PA317. The cell line transfected with a stably high expression of ribonuclease inhibitor was identified. We found that the transfected ribonuclease inhibitor could obviously inhibit cell proliferation, regulate cell cycle and induce cell apoptosis in vitro. Mice that were injected with the B16 cells transfected RI cDNA showed a significant inhibition of the tumor growth with lighter tumor weight, lower density of microvessels, longer latent periods, and survival time than those in the other two control groups. In conclusion, the results reveal the novel mechanism that antitumor effect of ribonuclease inhibitor is also associated with inducing apoptosis, regulating cell cycle and inhibiting proliferation besides antiangiogenesis. These results suggest that ribonuclease inhibitor might be a candidate of tumor suppressor gene in some tissues. RI could become a target gene for gene therapy. Our study may be of biological and clinical importance. PMID- 15778085 TI - Coenzyme Q10 affects expression of genes involved in cell signalling, metabolism and transport in human CaCo-2 cells. AB - Coenzyme Q10 is an essential cofactor in the electron transport chain and serves as an important antioxidant in both mitochondria and lipid membranes. CoQ10 is also an obligatory cofactor for the function of uncoupling proteins. Furthermore, dietary supplementation affecting CoQ10 levels has been shown in a number of organisms to cause multiple phenotypic effects. However, the molecular mechanisms to explain pleiotrophic effects of CoQ10 are not clear yet and it is likely that CoQ10 targets the expression of multiple genes. We therefore utilized gene expression profiling based on human oligonucleotide sequences to examine the expression in the human intestinal cell line CaCo-2 in relation to CoQ10 treatment. CoQ10 caused an increased expression of 694 genes at threshold-factor of 2.0 or more. Only one gene was down-regulated 1.5-2-fold. Real-time RT-PCR confirmed the differential expression for seven selected target genes. The identified genes encode proteins involved in cell signalling (n = 79), intermediary metabolism (n = 58), transport (n = 47), transcription control (n = 32), disease mutation (n = 24), phosphorylation (n = 19), embryonal development (n = 13) and binding (n = 9). In conclusion, these findings indicate a prominent role of CoQ10 as a potent gene regulator. The presently identified comprehensive list of genes regulated by CoQ10 may be used for further studies to identify the molecular mechanism of CoQ10 on gene expression. PMID- 15778087 TI - Chaperone-like activity of beta-casein. AB - The caseins are major components of milk for most mammals and are secreted as large colloidal aggregates termed micelles. They have less ordered secondary and tertiary structures in comparison with typical globular proteins. In this work, beta-casein, a member of the casein family, has been demonstrated to exhibit chaperone-like activity, being able to suppress the thermal and chemical aggregation of such substrate proteins as insulin, lysozyme, alcohol dehydrogenase, and catalase by forming stable complexes with the denaturing substrate proteins. Meanwhile, beta-casein was found to not only prevent aggregation of the substrate proteins, but also solubilize the protein aggregates already formed. Data also show that beta-casein exhibits a higher chaperone-like activity than alpha-casein, likely due to the difference in the number of proline residues present and/or in the extent of exposed hydrophobic surfaces. The implications for their in vivo functions of the caseins, based on their exhibiting such in vitro chaperone-like activities, are discussed. PMID- 15778088 TI - S100A9 deficiency alters adenosine-5'-triphosphate induced calcium signalling but does not generally interfere with calcium and zinc homeostasis in murine neutrophils. AB - The two calcium- and zinc-binding proteins, S100A9 and S100 A8, abundant in myeloid cells are considered to play important roles in both calcium signalling and zinc homeostasis. Polymorphonuclear neutrophils from S100A9 ko mice are also devoid of S100A8. Therefore, S100A9-deficient neutrophils were used as a model to study the role of the two S100 proteins in the neutrophils's calcium and zinc metabolism. Analysis of the intracellular zinc level upon pyrithione and (+/-) (E)-methyl-2-[(E)-hydroxyimino]-5-nitro-6-methoxy-3-hexeneamide (NOR-1) treatment revealed no differences between S100A9-deficient and wildtype neutrophils. Similar, the calcium signals were not distinguishable from S100A9-deficient and wildtype neutrophils upon stimulation with platelet activating factor (PAF), thapsigargin or macrophage inflammatory protein 1 alpha (MIP-1 alpha), indicating despite their massive expression S100A8/A9 do neither serve as calcium nor as zinc buffering proteins in granulocytes. In contrast, stimulation with adenosine 5'-triphosphate (ATP) induces a significant stronger increase of the intracellular free calcium level in S100A9-deficient cells compared to wildtype cells. Moreover, the ATP-induced calcium signal was still different when the cells were incubated in calcium free buffer suggesting that pirinergic receptors of the P(2Y) class could be involved in this signalling pathway. PMID- 15778089 TI - Expression of stress response protein glucose regulated protein-78 mediated by c Myb. AB - Glucose regulated protein-78, GRP78 has been implicated in the protection of tumor cells from cytotoxic damage and apoptosis. When protein profiles of colon cell lines were investigated we found remarkably high GRP78 expression in two cell lines. These cell lines express elevated levels of the transcription factor c-Myb due to genomic amplification of the c-myb locus and we hypothesized that c Myb regulates GRP78 expression in colon cancer cells. The promoters of human and murine GRP78 and the related family member GRP94 were examined and potential c Myb binding sites were identified and characterized. DNA binding studies with recombinant c-Myb and nuclear extracts together with ChIP assays on colon cell lines validated these sites. Endogenous GRP78 expression was further induced in these colon cells in response to Thapsigargin treatment, a potent inducer of the unfolded protein response. Transactivation studies with the human GRP78 promoter in colon cell lines showed reporter activity was dependent upon the presence of a conserved c-Myb binding site independent of sequences associated with the unfolded protein response. Finally, over-expression of c-Myb induced the endogenous GRP78 gene. These data suggest that amplification of c-myb in tumor cells may lead to robust GRP78 gene induction, which may in turn assist cells in survival under conditions of oxygen deprivation and nutrient stress. PMID- 15778091 TI - Beta-catenin/T-cell factor-mediated transcription is modulated by cell density in human bronchial epithelial cells. AB - The embryonic Wnt/beta-catenin ('canonical') pathway has been implicated in epithelial regeneration. To investigate the role of Wnt signal transduction in the airways, we characterised the expression of key pathway components in human bronchial epithelial cells (HBEC) and studied the influence of cell density on pathway activity, using sub-confluent cells in log-phase growth as a simple model of repairing epithelium. Primary HBEC and H292 bronchial epithelial cells were found to express TCF-4, TCF-3 and isoforms of LEF-1, transcription factors that are regulated by Wnt signalling. The cells also had the potential to respond to Wnt signalling through expression of several members of the Frizzled receptor family, including FZD-5 and -6. In confluent H292 cells, 20 mM lithium and 25% v/v Wnt-3a conditioned medium induced 4.5-fold (p = 0.008) and 1.4-fold (p = 0.006) increases in TOPflash activity, respectively. Under conditions of reduced cell density, TOPflash activity increased 1.8-fold (p = 0.002) in association with increased nuclear localisation of hypophosphorylated (active) beta-catenin and increased cell proliferation. This up-regulation in reporter activity occurred independently of EGF receptor activation and could not be recapitulated by use of low-calcium medium to disrupt cadherin-mediated cell-cell adhesion, but was associated with changes in FZD-6 expression. We conclude that reactivation of this embryonic pathway may play an important role in bronchial epithelial regeneration, and that modulation of Fzd-6 receptors may regulate Wnt signalling at confluence. Recognising that many chronic inflammatory disorders of the airways involve epithelial damage and repair, altered Wnt signalling might contribute to disease pathogenesis or progression. PMID- 15778090 TI - Contribution of L-3,4-dihydroxyphenylalanine metabolism to the inhibition of gluconeogenesis in rabbit kidney-cortex tubules. AB - The circulating L-3,4-dihydroxyphenylalanine, the drug of choice in the therapy of Parkinson's disease (PD), is efficiently extracted by kidney and converted to dopamine, known to control several renal functions. As: (i) in addition to liver, kidney is an important source of glucose in mammals and (ii) the action of this drug on renal gluconeogenesis has not yet been studied, the aim of the present investigation was to estimate the influence of L-3,4-dihydroxyphenylalanine metabolism on glucose formation in isolated kidney-cortex tubules incubated with various gluconeogenic substrates. The data indicate that a rapid intracellular degradation of L-3,4-dihydroxyphenylalanine and tyramine (at 100 and 200 microM concentrations) is accompanied by 25-40% decrease in glucose production from pyruvate, alanine + glycerol + octanoate and dihydroxyacetone due to augmented generation of hydrogen peroxide via monoamine oxidase B, resulting in a decline of glutathione redox state by 40%. Moreover, following inhibition of monoamine oxidase B by deprenyl or substitution of pyruvate by aspartate + glycerol + octanoate both L-3,4-dihydroxyphenylalanine and tyramine affect neither the rate of gluconeogenesis nor glutathione redox state. In view of: (i) L-3,4 dihydroxyphenylalanine- and tyramine-induced changes in intracellular levels of gluconeogenic intermediates, and (ii) a significant decline of phosphoenolpyruvate carboxykinase activity by 500 microM oxidized glutathione, it is likely that L-3,4-dihydroxyphenylalanine- and tyramine-evoked disturbances in the glutathione redox state might diminish flux through phosphoenolpyruvate carboxykinase and in consequence decrease glucose formation in renal tubules, suggesting a new potential side-action of L-3,4-dihydroxyphenylalanine treatment. PMID- 15778092 TI - A functional promoter region of the CKLFSF2 gene is located in the last intron/exon region of the upstream CKLFSF1 gene. AB - The genes for CKLFSF1 (chemokine-like factor super family member 1) and CKLFSF2 (chemokine-like factor super family member 2) are very closely linked, within 312 bp of each other. Here, we present evidence that the last intron/exon region of the CKLFSF1 gene contains a novel eukaryotic promoter capable of directing the expression of the downstream gene, CKLFSF2. We identified two segments of the upstream region of the CKLFSF2 gene, 2146 bp (-2134/+12, relative to ATG +1) and 1483 bp (-2134/-652), that were capable of efficiently driving expression of a linked reporter gene upon transient transfection into several kinds of cell lines. The 1483 bp segment exhibited more than a two-fold increase in luciferase activity relative to the 2146 bp segment. By analyzing 5'-deletion mutants of the 1483 bp segment, we identified a 195 bp segment (-846/-625) located in the last intron/exon region of the CKLFSF1 gene that was critical for promoter activity. DNA decoy experiments revealed that a 122 bp (-846/-725) fragment markedly inhibited CKLFSF2 mRNA transcription. Furthermore, we found that the putative promoter region of the CKLFSF2 gene is separated from the transcription start site by about 500 bp. Accumulating reports suggest that introns have many functions, including the modulation of regulation and structure. This work provides evidence that a eukaryotic gene promoter sequence from one gene located in an intron/exon of another. PMID- 15778093 TI - Apolipoproteins C-II and C-III inhibit selective uptake of low- and high-density lipoprotein cholesteryl esters in HepG2 cells. AB - Plasma low- and high-density lipoproteins (LDL and HDL) are cleared from the circulation by specific receptors and are either totally degraded or their cholesteryl esters (CE) are selectively delivered to cells by receptors such as the scavenger receptor class B type I (SR-BI). The aim of the present study was to define the effect of apoC-II and apoC-III on the uptake of LDL and HDL by HepG2 cells. Stable transformants were obtained with sense or antisense strategies that secrete 47-294% the normal level of apoC-II or 60-200% that of apoC-III. Different levels of secreted apoC-II or apoC-III had little effect on LDL and HDL protein degradation by HepG2 cells. However, compared to controls, cells under-expressing apoC-II showed a 160% higher capacity to selectively take up HDL-CE, while cells under-expressing apoC-III demonstrated 70 and 160% higher capacity to take up CE from LDL and HDL, respectively. In experiments conducted with exogenously added apoC-II or apoC-III, no significant effect was observed on lipoprotein-protein association/degradation; however, LDL-CE and HDL-CE selective uptake was significantly reduced in a dose-dependent manner. These results indicate that apoC-II and apoC-III inhibit CE-selective uptake. PMID- 15778094 TI - Hypoxia and recovery perturb free radical processes and antioxidant potential in common carp (Cyprinus carpio) tissues. AB - The effects of hypoxia exposure and subsequent normoxic recovery on the levels of lipid peroxides (LOOH), thiobarbituric acid reactive substances (TBARS), carbonylproteins, total glutathione levels, and the activities of six antioxidant enzymes were measured in brain, liver, kidney and skeletal muscle of the common carp Cyprinus carpio. Hypoxia exposure (25% of normal oxygen level) for 5h generally decreased the levels of oxidative damage products, but in liver TBARS content were elevated. Hypoxia stimulated increases in the activities of catalase (by 1.7-fold) and glutathione peroxidase (GPx) (by 1.3-fold) in brain supporting the idea that anticipatory preparation takes place in order to deal with the oxidative stress that will occur during reoxygenation. In liver, only GPx activity was reduced under hypoxia and reoxygenation while other enzymes were unaffected. Kidney showed decreased activity of GPx under aerobic recovery but superoxide dismutase (SOD) and catalase responded with sharp increases in activities. Skeletal muscle showed minor changes with a reduction in GPx activity under hypoxia exposure and an increase in SOD activity under recovery. Responses by antioxidant defenses in carp organs appear to include preparatory increases during hypoxia by some antioxidant enzymes in brain but a more direct response to oxidative insult during recovery appears to trigger enzyme responses in kidney and skeletal muscle. PMID- 15778095 TI - Substantial decrease of urinary 8-oxo-7,8-dihydroguanine, a product of the base excision repair pathway, in DNA glycosylase defective mice. AB - Genome integrity is maintained via removal (repair) of DNA lesions and an increased load of such DNA damage has been linked to numerous pathological conditions, including carcinogenesis and ageing. 8-Oxo-7,8-dihydroguanine is one of the most critical lesions of this type. The free 8-oxo-7,8-dihydroguanine produced by the action of a specific DNA glycosylase is a potential source of this compound in urine. To date, there has been no direct, experimental evidence demonstrating that urinary 8-oxo-7,8-dihydroguanine is produced by the base excision repair pathway. For clarification of this issue, we applied a recently developed methodology which involved high performance liquid chromatography pre purification followed by gas chromatography with isotope dilution mass spectrometric detection to compare the urinary excretion rate of 8-oxo-7,8 dihydroguanine in wild type and OGG1 glycosylase knock out mice. Our study revealed a 26% reduction in urinary level of 8-oxo-7,8-dihydroguanine in OGG1 deficient mice in comparison with the wild type strain. This clearly indicates that the mouse OGG1 glycosylase contributes significantly to the generation of urinary 8-oxo-7,8-dihydroguanine. Therefore, urinary measurements of 8-oxo-7,8 dihydroguanine may be attributed to DNA damage and repair, which in turn suggests that they may be useful in studying associations between DNA repair and disease. PMID- 15778096 TI - Stroke thrombolysis: we need new data, not more reviews. PMID- 15778097 TI - Treatment for the progression of Parkinson's disease. PMID- 15778098 TI - Pregabalin for painful neuropathy. PMID- 15778099 TI - Interferon beta, progressive MS, and brain atrophy. PMID- 15778100 TI - The neuro-ophthalmology of multiple sclerosis. PMID- 15778101 TI - Neurotoxic marine poisoning. AB - Marine poisoning results from the ingestion of marine animals that contain toxic substances and causes substantial illness in coastal regions. Three main clinical syndromes of marine poisoning have important neurological symptoms-ciguatera, tetrodotoxin poisoning, and paralytic shellfish poisoning. Ciguatera is the commonest syndrome of marine poisoning and is characterised by moderate to severe gastrointestinal effects (vomiting, diarrhoea, and abdominal cramps) and neurological effects (myalgia, paraesthesia, cold allodynia, and ataxia), but is rarely lethal. Tetrodotoxin poisoning and paralytic shellfish poisoning are less common but have a higher fatality rate than ciguatera. Mild gastrointestinal effects and a descending paralysis are characteristic of these types of poisoning. In severe poisoning, paralysis rapidly progresses to respiratory failure. Diagnosis of all types of marine poisoning is made from the circumstances of ingestion (type of fish and location) and the clinical effects. Because there are no antidotes, supportive care, including mechanical ventilation in patients with severe paralysis, is the mainstay of treatment. PMID- 15778102 TI - Assessment of disease progression in motor neuron disease. AB - Motor neuron disease (MND) is characterised by progressive deterioration of the corticospinal tract, brainstem, and anterior horn cells of the spinal cord. There is no pathognomonic test for the diagnosis of MND, and physicians rely on clinical criteria-upper and lower motor neuron signs-for diagnosis. The presentations, clinical phenotypes, and outcomes of MND are diverse and have not been combined into a marker of disease progression. No single algorithm combines the findings of functional assessments and rating scales, such as those that assess quality of life, with biological markers of disease activity and findings from imaging and neurophysiological assessments. Here, we critically appraise developments in each of these areas and discuss the potential of such measures to be included in the future assessment of disease progression in patients with MND. PMID- 15778103 TI - Progressive myoclonic epilepsies: a review of genetic and therapeutic aspects. AB - The progressive myoclonic epilepsies (PMEs) are a group of symptomatic generalised epilepsies caused by rare disorders, most of which have a genetic component, a debilitating course, and a poor outcome. Challenges with PME arise from difficulty with diagnosis, especially in the early stages of the illness, and further problems of management and drug treatment. Recent advances in molecular genetics have helped achieve better understanding of the different disorders that cause PME. We review the PMEs with emphasis on updated genetics, diagnosis, and therapeutic options. PMID- 15778105 TI - Assessment of professional development of neurology trainees: a UK perspective. AB - The European Working Time Directive has led to concerns that trainees' hours of work will decrease, with negative effects on their training. Various changes to the curriculum are needed to equip trainees for self-directed learning. A strong lobby argues for the introduction of examinations before entry to the consultant grade. Meanwhile, there is intense political pressure (in the UK at least), to shorten the duration of postgraduate training so that pledges for consultant numbers can be met. Reconciliation of these conflicting demands is a challenge for the future. Additional assessment procedures should be piloted. To successfully introduce changes, trainees and trainers will need to work in partnership. Course assessment and student feedback should be formalised. In this paper I summarise the assessment procedures in the UK and US, and suggest a model of best practice. PMID- 15778104 TI - Alteplase and ischaemic stroke: have new reviews of old data helped? AB - BACKGROUND: Thrombolysis for stroke is still not widely used as current recommendations restrict treatment to selected patients. In general, these are patients who can be assessed quickly by specialised stroke teams, have intracranial haemorrhage excluded by appropriate brain imaging, and are treated with alteplase (recombinant tissue plasminogen activator; rt-PA) within 3 h of symptom onset. There is, however, still much debate regarding the scope of treatment and the reorganisation of services required to support an effective service. RECENT DEVELOPMENTS: Two recent publications have helped clarify some issues. The first was an individual-patient data meta-analysis of the alteplase trials. These analyses suggest treatment effects beyond the usual 3 h time window, but other than time to treatment no other factors influenced the effects of treatment. The second publication was a reanalysis of the original National Institute of Neurological Disorders and Stroke (NINDS) alteplase trial, done after criticism of the original study. The reanalysis confirmed that there was significant baseline imbalance of stroke severity between treatment and control groups in the NINDS trial, but established that this did not materially affect the positive results of the trial. However, the recording of blood pressure in the study was found to be inconsistent and therefore unsuitable for reanalysis. The previously published data on recommendations for blood-pressure control, arising from the NINDS trial, needs to be reconsidered in this light. Both studies included too few patients to provide reliable data on which clinical and radiological features influence the response to alteplase. WHERE NEXT?: The individual-patient data meta-analysis and reanalysis of the NINDS trial have probably exhausted the potential of previous trials to answer questions on the effects of thrombolysis. Further randomised trials comparing thrombolysis with control will be required to determine whether elderly people benefit from treatment or whether there are worthwhile benefits from alteplase beyond 3 h (and in such patients, whether advanced magnetic resonance imaging is an effective way to select those most likely to benefit). Various new approaches to reperfusion also require assessment in large-scale trials: new thrombolytic drugs, the combination of intravenous and intra-arterial thrombolytic drugs, combinations of thrombolytics with new antiplatelet agents, and augmentation of thrombolysis either with mechanical devices or with transcranial ultrasound. PMID- 15778106 TI - Neurology residency training in Europe: an Italian perspective. AB - Education is the most important part of medical-science training. Assessment of training programmes and examination of trainees' skills are necessary to guarantee that trainees develop competence. Training programmes vary between and within countries. The impending integration of the medical-job market among European countries highlights the need for people training to be specialists to achieve a certain standard. Here, we review the neurological training programmes in Europe, principally those in Italy, and discuss the standardisation of the training of residents in Italy and the quality control of current residency training, or its absence, in Europe. PMID- 15778108 TI - Management of human immunodeficiency virus infection in pregnancy. AB - The HIV global epidemic is having a devastating effect on women of reproductive age; women aged 15-24 years are 2.5 times more likely to be infected than young men in the same age group. Further, mother-to-child transmission (MTCT) accounts for almost two-thirds of the new infections that occur in children world-wide, annually. MTCT of HIV-1 varies widely and is dependent on obstetric practices, mode of delivery, breastfeeding, and the level of the viral load in the mother. Antiretroviral therapy (ARV) in pregnancy is prescribed for two main reasons: (i) women who need ARV medication for their own health; (ii) women who do not need treatment, or do not have access to treatment are offered prophylaxis to prevent MTCT, using one of a number of ARV regimens known to be effective. HIV infection is also associated with significant maternal morbidity and mortality. Clinicians caring for HIV-infected women need to update their knowledge continuously to provide optimal care. PMID- 15778109 TI - Breastfeeding and HIV. AB - Breastfeeding is a route of HIV transmission from an HIV-infected mother to her infant. However, breastfeeding is an important pillar of child survival and the ideal way of feeding an infant, as well as providing a unique biological and emotional basis for child development. This chapter will highlight the dilemma created by the risks and benefits of breastfeeding and will discuss factors which increase the risk of HIV transmission during breastfeeding, as well as strategies which could be employed to reduce these risks. The research agenda is very full as many questions still remain unanswered. PMID- 15778110 TI - Economic impact of HIV. AB - Economics affects everyone and the economic impact of HIV/AIDS will take generations to unfold. Recent macroeconomic predictions have emphasized the concept of human capital, and predicted that much higher costs will be associated with the epidemic than earlier suggestions that economies might simply grow more slowly implied. At a firm or sector level AIDS increases the costs of doing business; impact studies at this level are therefore more important for planning. Impacts at household level are some of the most immediate and devastating, owing to the close relationship between households being affected by HIV/AIDS and subsequent impoverishment. The economic impacts of HIV/AIDS at each level will be modified through prevention and treatment interventions. The potential savings to governments through implementation of a programme to prevent mother-to-child transmission provide some of the most compelling evidence for action. However, health services are suffering under a dual burden of increased demand and reduced capacity to deliver, which contributes to a cycle of poor health and economic decline. PMID- 15778111 TI - Adaptive evolution in perinatal HIV-1. AB - The immune-viral dynamics of the transmission of HIV-1 from mother to child are poorly understood, despite 20 years of research. Here we review evidence that the maternal immune response against HIV-1 can select forms of the virus that evade immunity and when transmitted have negative consequences in the child. Moreover, recent studies indicate that when wild-type virus is transmitted, an early immune response in the child can lead to the selection of viral escape forms in the first few months of life. These data suggest that adaptive immune surveillance in both mother and child contributes to the pathogenesis of early perinatal HIV-1. These observations augment our general understanding of the processes that determine the evolution of HIV-1 as it passes from one host to another. PMID- 15778112 TI - HIV and gynaecological infections. AB - Human immunodeficiency virus (HIV) infection primarily affects women during their reproductive years, and the co-existence of gynaecological infections is not surprising, given the fact that HIV is mainly acquired via heterosexual contact. Most gynaecological infections are themselves sexually acquired, and have the potential to increase the risk both of acquiring and transmitting the HI virus. As most sexually transmitted infections are asymptomatic, there is a need to improve methods of diagnosis and algorithms for early detection of sexually transmitted infections. HIV infection, however, particularly advanced disease, may alter the clinical presentation, course and response to conservative treatment for some of the sexually transmitted infections. PMID- 15778113 TI - Laboratory markers associated with progression of HIV infection. AB - Infection with HIV may develop to AIDS at different rates in different individuals, with a spectrum varying from rapid progression to long-term non progression. The variable course of HIV-1 infection causes emotional trauma for the infected person and complicates the design and interpretation of therapeutic trials because of unrecognized differences in prognosis. Owing to the variable clinical expression of HIV infection, the use of non-clinical disease markers has become important to patient management. Thus, it is essential to have tests which can accurately assess the stage of infection in an individual, as well as predict its course and monitor its progression. These laboratory tests are valuable during the period of clinical latency and subsequently supplement various clinical parameters. PMID- 15778114 TI - Ethical issues in HIV. AB - The number of people with HIV/AIDS continues to increase globally. Women, who represent the subgroup with the fastest rate of increase, are usually informed of their serostatus by the obstetrician/gynaecologist. As treatment of infected women raises a number of ethical issues, an understanding of the theoretical background for ethical decision making is requisite to ensure these problems are resolved within a morally appropriate framework. Vigorous debate has arisen from the tensions between the competing goals of HIV testing, third party disclosure, management of the critically ill HIV-infected woman, infertility management in the background of HIV/AIDS, and gender-based violence as cause or result of acquiring HIV infection. Women may be differently empowered economically, socially and culturally. What may be a satisfactory solution in the context of the USA and Europe may be far from ideal in that of the developing world. PMID- 15778115 TI - HIV and cancer of the cervix. AB - Cancer of the cervix is the second most common cause of cancer-related death in women worldwide, and in some low resource countries accounts for the highest cancer mortality in women. The highest burden of the HIV/AIDS epidemic is currently in sub-Saharan Africa, where more than half of the people infected are women who have no access to cervical cancer screening. The association between HIV and invasive cervical cancer is complex, with several studies now clearly demonstrating an increased risk of pre-invasive cervical lesions among HIV infected women. However, there have not been significantly higher incidence rates of invasive cervical cancer associated with the HIV epidemic. The highest numbers of HIV-infected women are in poorly-resourced countries, where the natural progression of HIV disease in the absence of highly active antiretroviral treatment sometimes results in deaths from opportunistic infections before the onset of invasive cervical cancer. This chapter will discuss the association of HIV and cervical intraepithelial neoplasia, the treatment of pre-invasive lesions, and invasive cervical cancer in HIV-infected women. The role of screening and the impact of antiretroviral treatment on the progression of pre invasive and invasive cancer will also be discussed. PMID- 15778116 TI - The challenges of HIV vaccine development and testing. AB - A vaccine against HIV remains the best hope for bringing the epidemic under control. An intensive global effort is underway to develop such a vaccine; however, the challenges are considerable. Several new vaccine technologies that have been developed and shown promise in animal models are now being tested in early phase safety trials in humans. Because there is no laboratory assay that will predict whether an HIV vaccine can protect humans from infection, clinical trials involving thousands of volunteers will need to be conducted to determine the efficacy of HIV vaccines. These trials need to take place in the developing countries that bear the burden of the epidemic, requiring a substantial amount of infrastructure development and capacity building. PMID- 15778117 TI - Characterization and immunomodulating activities of polysaccharide from Lentinus edodes. AB - The polysaccharide L-II was isolated and purified from the fruiting body of Lentinus edodes, which consisted of d-glucopyranose and had the molecular weight of 2.03 x 10(5) Da. We evaluated the effects of the polysaccharide L-II on the cellular immune response of Sarcoma 180-bearing mice. Mice were treated with three doses of the polysaccharide L-II (1, 5, and 10 mg/kg body weight) for 10 days. Tumor weight, relative spleen and thymus weight, delayed-type hypersensitivity (DTH) response, phagocytosis of macrophage, splenocytes proliferation were studied. Concentration of tumor necrosis factor-alpha (TNF alpha), interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) in mice serum were measured in control and polysaccharide groups. At the dose of 1, 5 and 10 mg/kg, a significant increase (p<0.05) in relative spleen and thymus weight, DTH, phagocytosis of macrophage was observed, as well as a significant decrease in tumor formation. The concentration of TNF-alpha, IFN-gamma in serum increased significantly in the polysaccharide groups compared with the model control group, but IL-2 not. Moreover, the polysaccharide L-II could increase NO production and catalase activity in macrophages. Results of these studies demonstrated the antitumor activity of the polysaccharide L-II on mice-transplanted sarcoma 180 was mediated by immunomodulation in inducing T-cells and macrophage-dependent immune system responses. PMID- 15778118 TI - Involvement of P-gp in the process of apoptosis in peripheral blood mononuclear cells. AB - Multidrug resistance mediated by the drug-efflux protein P (P-gp) is one of mechanisms that cells use to escape death induced by drugs and other agents. The aim of the study was to evaluate the effect of P-gp inhibition on apoptosis of PHA-activated peripheral blood mononuclear cells (MNC) as well as apoptosis induced by methotrexate (MTX), dexamethasone (DEX), methylprednisolone (MP) and cortisone (COR). Apoptosis was quantified by flow cytometry using Annexin V/PI and terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL). P-gp expression was inhibited using verapamil (VER) and P-gp specific monoclonal antibodies (mAb). VER and mAb enhanced the apoptosis of PHA-activated MNC. Moreover these agents significantly increased the apoptosis induced by MTX, DEX, MP and COR. The results of this study suggest that P-gp is involved in the process of apoptosis in peripheral blood mononuclear cells. PMID- 15778119 TI - Effect of D-limonene on immune response in BALB/c mice with lymphoma. AB - The monoterpene D-limonene and its metabolites have been shown to exert chemopreventive and chemotherapeutic activities against different tumors in animal models and clinical trials. However, it is unknown whether these compounds modulate the immune response in tumor-bearing mice. We evaluated the survival of lymphoma-bearing mice fed with a diet with D-limonene. To assess the cell immune response, we sensitized and challenged BALB/c mice with 2,4-dinitrofluorobenzene (DNFB) and evaluated the T-cell subpopulations by flow cytometry. We also examined phagocytosis, microbicidal activity and chemotactic function in peritoneal macrophages. In order to know the role of D-limonene and its metabolites, macrophage NO production and lymphocyte proliferation studies were performed in vitro with D-limonene, perillic acid and perillyl alcohol. The results showed that D-limonene increased the survival of lymphoma-bearing mice, delayed hypersensitivity reaction to DNFB, phagocytosis and microbicidal activity. In vitro studies indicate that D-limonene increased NO production in peritoneal macrophages obtained from tumor-bearing mice. Our data suggest that in addition to reported properties, D-limonene modulates the immune response with significant potential for clinical application. PMID- 15778120 TI - Differential roles of constitutively activated ERK1/2 and NF-kappa B in cytotoxicity and proliferation by human NK cell lines. AB - Compared with freshly isolated peripheral blood natural killer (NK) cells, the YT and NK-92 cell lines are characterized by elevated cytolytic activity. The molecular mechanisms underlying the rapid proliferation and enhanced lytic activity of NK cell lines are poorly understood. Investigation of these cell lines revealed that ERK1/2 and NF-kappa B are constitutively activated, providing evidence that these two signaling pathways are differentially involved in cytolysis and proliferation. Furthermore, blocking ERK1/2 activation with the specific inhibitor, PD098059, inhibited cytolytic activity in both cell lines and reduced mRNA expression of cytolysis-related effector molecules such as Fas-L and IFN gamma, as measured by semi-quantitative RT-PCR. However, MTT colormetric analysis showed that treatment with the PD098059 inhibitor did not affect cell proliferation. Meanwhile, blockade of the NF-kappa B signaling pathway using MG132 inhibited cellular growth without impacting cytolytic capability. No synergistic interactions were observed between ERK1/2 and NF-kappa B after combination treatment with PD098059 and MG132, suggesting that these two signaling pathways likely affect cellular proliferation and cytotoxicity by NK cells differentially. PMID- 15778121 TI - Plant-derived anti-inflammatory compounds affect MIF tautomerase activity. AB - The cytokine macrophage migration inhibitory factor (MIF) has recently emerged as a crucial factor in the pathogenesis of rheumatoid arthritis (RA). It is debated whether the MIF mediated tautomeric conversion of either phenylpyruvate or of its other phenolic substrates is implicated in the pro-inflammatory action of this cytokine. Traditional herbal remedies have been used for centuries to alleviate inflammatory ailments of many kinds including arthritis. Several of their active ingredients identified are mono- or poly-phenol derivatives. In the present study the effect of some anti-inflammatory plant phenols on MIF mediated tautomerism of phenylpyruvate was investigated. Curcumin and caffeic acid were found to be the most potent inhibitors, exhibiting IC(50) values in the submicromolar range in the ketonase assay. Resveratrol and umbelliferon were almost as potent inhibitors as the antipyretic-analgetic drug acetaminophen. Our results reveal MIF as a possible target for the herbal anti-rheumatic agents. PMID- 15778122 TI - Cells isolated from bone-marrow and lungs of allergic BALB/C mice and cultured in the presence of IL-5 are respectively resistant and susceptible to apoptosis induced by dexamethasone. AB - We have previously reported that, in IL-5-stimulated bone-marrow cultures, dexamethasone upregulates eosinophil differentiation and protects developing eosinophils from apoptosis induced by a variety of agents. Recently developed procedures for the isolation of hemopoietic cells from allergic murine lungs have enabled us to evaluate how these cells respond to dexamethasone in IL-5 stimulated cultures, when compared with bone-marrow-derived cells isolated from the same donors, and whether differences in response patterns were linked to apoptosis. Ovalbumin challenge of sensitized mice increased significantly the numbers of mature leukocytes as well as hemopoietic cells recovered from digested lung fragments, relative to saline-challenged, sensitized controls. Both mature eosinophils and cells capable of differentiating into eosinophils in the presence of IL-5 were present in lungs from sensitized mice 24 h after airway challenge. Dexamethasone strongly inhibited eosinophil differentiation in IL-5-stimulated cultures of lung hemopoietic cells. By contrast, dexamethasone enhanced eosinophil differentiation in cultures of allergic bone-marrow cells, in identical conditions. Hemopoietic cells from lungs and bone-marrow were respectively susceptible and resistant to induction of apoptosis by dexamethasone. The dexamethasone-sensitive step was the response to IL-5 in culture, while accumulation of IL-5 responsive cells in allergen-challenged lungs was dexamethasone-resistant. Cells from lungs and bone-marrow, cultured for 3 days with IL-5 in the absence of dexamethasone, did not respond to a subsequent exposure to dexamethasone in the presence of IL-5. These findings confirm that IL 5-responsive hemopoietic cells found in challenged, sensitized murine lungs differ from those in bone-marrow, with respect to the cellular responses induced by dexamethasone, including apoptosis. PMID- 15778123 TI - Juzentaihoto, a Kampo medicine, enhances IL-12 production by modulating Toll-like receptor 4 signaling pathways in murine peritoneal exudate macrophages. AB - Juzentaihoto (TJ-48), a Kampo medicine, has been reported to affect the immune system. Although toll-like receptors (TLRs) have been identified as receptors of innate immunity, the effects of TJ-48 on TLR signaling pathways have not been thoroughly investigated. Here we evaluated the effects of TJ-48 on TLR4 signaling pathways. Peritoneal exudate macrophages (PEMs) isolated from mice orally administered TJ-48 for 11 days were stimulated with lipopolysaccharide (LPS), a ligand of TLR4, in vitro. Production of IL-12 p40 was significantly augmented in TJ-48-treated PEMs compared with that in vehicle PEMs, without affecting the surface expression of TLR4. Treatment with chemical inhibitors of NF-kappa B and p38 mitogen-activated protein kinases (MAPKs) in vitro inhibited LPS-induced IL 12 production, whereas JNK and ERK inhibitors increased IL-12 production. Immunoblotting with phosphorylation-state specific antibodies demonstrated that TJ-48 differentially affected LPS-induced phosphorylation of NF-kappa B and MAPKs. In PEMs treated with TJ-48, LPS-induced phosphorylation of p65 NF-kappa B and p38 MAPK was augmented, while that of JNK and ERK was attenuated compared with those in vehicle PEMs. These results suggest that selective modulation of the TLR4 signaling pathways by TJ-48 is involved in enhanced production of IL-12 in PEMs. PMID- 15778124 TI - A potent and selective inhibitor of group IIa secretory phospholipase A2 protects rats from TNBS-induced colitis. AB - Secretory phospholipase A(2) (sPLA(2)) enzymes have been implicated in the pathogenesis of human inflammatory bowel disease (IBD). In this study we compared the efficacy of a potent, new and highly selective inhibitor of group IIa human sPLA(2) enzyme (5-(4-benzyloxyphenyl)-4S-(7-phenylheptanoylamino)-pentanoic acid; sPLA(2)I), with that of sulfasalazine, in a rat model of trinitrobenzene sulfonic acid (TNBS)-induced colitis. Following a single oral dose of sPLA(2)I (5 mg/kg), pharmacoactive levels of drug were detected in the serum within 15 min and for up to 24 h by liquid chromatography mass spectrometry analysis. Rats treated with sPLA(2)I (5 mg/kg/day) prior to induction of colitis were significantly healthier than TNBS-alone rats, as shown by reduced mortality, improved food intake and increased body weight, and significantly reduced colon myeloperoxidase levels, edema, tumour necrosis factor-alpha levels, and colon macroscopic pathology scores after 8 days. Rats pretreated with sulfasalazine (100 mg/kg/day) also had reduced disease expression markers similar to the sPLA(2)I, but exhibited no improvement in colon edema. This study supports a role for the group IIa sPLA(2) enzyme in pathology associated with the TNBS rat model of IBD, and suggests a possible therapeutic application for selective inhibitors of group IIa sPLA(2) inhibitors in the treatment of IBD. PMID- 15778125 TI - Effect of Candida albicans septicemia on the cardiovascular function of rabbits. AB - Candida albicans is an opportunistic pathogen that causes life-threatening systemic infection in immunocompromised host. However, little is known about the effects of yeast on the cardiovascular functions. This study examined the effects of C. albicans septicemia on the heart and vessel functions and nitric oxide (NO) production in infected rabbits. Anaesthetized animals were challenged with intravenous C. albicans (6 x 10(8)/kg) or saline and the blood pressure of rabbits were measured over 5 h. After that response of the isolated thoracic aorta, right atrium and left papillary muscle were recorded. Blood pressure significantly decreased in the infected rabbits during the septicemia but in the control animals it was stable. The blood nitrite levels and NO-synthases (eNOS, iNOS) expression and tissue nitrite levels in the heart and aorta were similar in the both groups. In the aorta isolated from C. albicans-infected rabbits, acetylcholine-induced endothelium-dependent relaxation was decreased, but contractions induced by phenylephrine were potentiated. The NOS inhibitor, L-N(G) nitro-arginine methyl ester (L-NAME)-induced contraction increase in the right atrium was depressed by the yeast-infection. In the heart and aorta, microscopic examination revealed no tissue invasion of C. albicans. These results indicate the ability of C. albicans-induced septicemia to destroy NO-related responses of the heart and aorta and may have important implications for functional damage to endothelium and the regulation of cardiovascular functions. In addition, NOS induction and NO over-production are not stimulated by systemic C. albicans infection, which would alter the host immune reaction and homeostasis. PMID- 15778126 TI - The liquid culture filtrates of Paecilomyces tenuipes (Peck) Samson (=Isaria japonica Yasuda) and Paecilomyces cicadae (Miquel) Samson (=Isaria sinclairii (Berk.) Llond) regulate Th1 and Th2 cytokine response in murine Peyer's patch cells in vitro and ex vivo. AB - The effects of liquid culture filtrates of medicinal entomogenous fungi, Paecilomyces tenuipes (Peck) Samson (=Isaria japonica Yasuda or Isaria tenuipes) (PTCF) and Paecilomyces cicadae (Miquel) Samson (=Isaria sinclairii (Berk.) Llond) (PCCF), on cytokine productions in cultured Peyer's patches (PP) from C57BL/6J mice were investigated in vitro and ex vivo. In an in vitro experiment, PTCF (100 and 10 microg/ml) enhanced the production of T helper 1 (Th1) cytokines, interleukin (IL)-2 and interferon (IFN)-gamma, in cultured PP cells stimulated with 5 microg/ml concanavalin A (Con A) but did not influence on the production of T helper 2 (Th2) cytokines, IL-4 and IL-5. PTCF also enhanced the production of granulocyte macrophage colony-stimulating factor (GM-CSF) and IL-10 in the cultured PP cells. While, PCCF enhanced the production of IFN-gamma but did not alter the level of IL-2 in the PP cells. In an ex vivo experiment using PP cells removed from the mice after oral treatment of PTCF (10 and 100 mg/kg daily for 7 consecutive days), the production of IL-2 and IFN-gamma were increased in response to Con A. On the other hand, orally treated PCCF (10 mg/kg/day) suppressed IL-2 production but did not change the levels of IFN-gamma and IL-10 in the isolated PP cells. The flow cytometric analysis revealed that the population of CD3(+) cells in the PP cells slightly but significantly increased after oral administration of PCCF. Orally administered PTCF did not change the population of T (CD3(+)), B (CD19(+)), T cell subset (CD4(+)and CD8(+)) and Th1 (IFN-gamma(+)) and Th2 (IL-4(+)). From PTCF, the fraction rich in proteoglycans was separated as active fraction that stimulates Th1 immune response. These results indicate that the mode of action of PTCF and PCCF on mucosal immune response is different and this is contributed to their metabolites. Taken together, there is a possibility of PTCF and PCCF being therapeutic or preventive agents for immune diseases such as cancer, allergy and parasitic disease through activation of mucosal immune response. PMID- 15778127 TI - Effects of a short-course of amoxicillin/clavulanic acid on systemic and mucosal immunity in healthy adult humans. AB - Although amoxicillin/clavulanic acid (AMC) is the most frequently administered antibiotic in France, its in vivo effects on immunity in healthy adults have never, to our knowledge, been described. Eighteen healthy adult male volunteers, 25+/-6 years old, were treated for 5 days with oral amoxicillin (1 g) /clavulanate potassium (125 mg), two times daily. Systemic and local intestinal immunity parameters were sequentially explored before, during and after the antibiotic treatment. No significant differences were obtained for transudation markers (albumin and alpha1-antitrypsin) in sera, feces and saliva, showing that AMC did not induce inflammatory reaction. Phagocytosis, peripheral blood cell subsets, intracellular interferon (IFN)-gamma and tumor necrosis factor (TNF) alpha production by natural killer (NK) cells and cytotoxic T lymphocytes, intracellular TNF-alpha production by monocytes showed no significant differences throughout the trial. In fecal outputs, no significant differences were found in secretory immunoglobulin A (S-IgA), lactoferrin (Lf), lysozyme (Lz) and transforming growth factor (TGF)-beta1. In sera, concentrations of total IgA (T IgA), S-IgA, IgM, Lf and Lz did not show any significant variations throughout the study, whereas concentrations of IgG were slightly but significantly reduced 15 days after AMC treatment. In saliva, concentrations of T-IgA were slightly but significantly higher, whereas S-IgA concentrations were unchanged. Our results showed that oral AMC intake did not induce any significant adverse effects on immunity in adult humans. PMID- 15778129 TI - Correlates of strength training in older rural African American and Caucasian women. AB - This study examined factors influencing strength training (ST) in two convenience samples of older rural women. Focus group (FG) participants were 23 Caucasian and 16 African American women aged 67.5 +/- 9.2 years. Survey participants were 60 Caucasian and 42 African American women, aged 70.59 +/- 9.21 years. FG participants answered questions about the risks, benefits, and barriers to ST. Survey participants completed measures of demographics, physical activity (including ST), depression and stress, decisional balance for exercise (DBE), barriers to PA, and social support (SS). Regression modeling examined correlates of ST. FG participants identified physical health gains and improved appearance as ST benefits. African American women also included mental health benefits and "feeling good". Both Caucasian and African American groups named physical health problems as risks of ST. Caucasian women identified time constraints, lack of ST knowledge, physical health problems, lack of exercise facilities, and the cost of ST as barriers. African American women cited being "too tired", physical health problems, lack of support, and other family and work responsibilities. The linear regression model explained 23.2% of the variance in hours per week of ST; DBE and family SS were independent positive correlates. This study identified correlates to participation in ST in older rural women and provides a basis for developing ST interventions in this population. PMID- 15778128 TI - Effect of a fermented ginseng extract, BST204, on the expression of cyclooxygenase-2 in murine macrophages. AB - This paper investigates how BST204, a fermented ginseng extract, affects the expression and mechanism of cyclooxygenase-2 (COX-2). BST204 was prepared by incubating crude ginseng extract with ginsenoside-beta-glucosidase. Unexpectedly, BST204 had no effect on the level of COX-2 protein in unstimulated RAW 264.7 cells, and it suppressed the level of COX-2 protein and PGE(2) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. It did not show any suppressive effect, though, on the COX-2 mRNA level. To investigate the suppressive mechanism of COX-2 protein, the activating phosphorylation of p70 S6 kinase and 4E-BP1, which are important for translation, were measured. The phosphorylation of p70 S6 kinase, not 4E-BP1, was increased by LPS in a time dependent manner, and was inhibited by BST204 in a dose-dependent manner. The expression of COX-2 protein, however, was partially suppressed by rapamycin, an upstream inhibitor of p70 S6 kinase. Therefore, this paper suggests that the suppression of COX-2 protein by BST204 was partially correlated with the inhibition of p70 S6 kinase activation. PMID- 15778130 TI - Popular health promotion strategies among Chinese and East Indian immigrant women. AB - PURPOSE: To advance understanding about the popular health promotion strategies and factors associated with the successful transfer and uptake of health messages among Chinese and Indian immigrant women. METHODS: Eight focus groups were conducted with 46 immigrant women, 24 from Mainland China and 22 from India, who had lived less than 5 years in Canada. Audiotaped data were transcribed, translated and analyzed by identification of themes and subcategories within and between groups. RESULTS: In both ethnic groups, discussions on promoting health messages had five major themes, i.e., sources, barriers, facilitators, credibility and ways to improve access along with group specific sub-themes. Despite identification of several diverse sources of health information in the adopted country, Indian and Chinese immigrant women perceived most strategies as not very effective. The reasons of perceived ineffectiveness were barriers to accessing and comprehending the health messages; and limited prior exposure to institution based or formal health promotion initiatives. These women were more familiar with informal means of obtaining health information such as social networks, mass media and written materials in their mother tongue. CONCLUSION: Existing health communication and health promotion models need to be re orientated from a one-way information flow to a two-way dialogue model to bridge the gap between program efficacy and effectiveness to reach underserved immigrant women. An "outside the box" approach of non-institutional informal health promotion strategies needs to be tested for the studied groups. PMID- 15778131 TI - Universal prenatal HIV screening: patient attitudes and perceptions. AB - In 1995, Center for Disease Control guidelines were modified to replace targeted HIV screening of pregnant women with universal screening. Public health scholars have argued that one rationale for universal screening is to reduce the potential stigmatization of women of color and lower socioeconomic status who are typically targeted under selective screening. Here, we examine whether experiences and impressions of prenatal screening differ across socioeconomic and racial lines. Data are drawn from an NIH-funded pilot study of 353 pregnant women across several sites in Connecticut, a state with unusually strong prenatal HIV screening laws. We find no significant differences on testing protocols and attitudes toward mandatory HIV screening. However, minority and lower socioeconomic status women are more likely (1) to understand the screening law, (2) to report that their doctor stresses the importance of the test, and (3) to express much higher levels of concern about the confidentiality of the test. PMID- 15778132 TI - Women's approaches to the use of new technology for cancer risk information. AB - BACKGROUND: Women are currently offered multiple sources of breast cancer risk information, and the Internet is the most rapidly growing health information source. The objectives of this study were to identify women's preferences for receiving online breast cancer risk information, to identify barriers to accessing this information, and to identify differences in these factors between Internet users and non-users. METHODS: Eight focus groups were conducted with women aged 18 to 74 (n = 65) living in King County, Washington. RESULTS: Participants were interested in accessing online health information with the support of a real person to assist with information searches and in accessing personalized information online. Participants emphasized the importance of trust, both in the content provider and in the organization collecting their personal data. CONCLUSIONS: Web sites should provide personalized information and personal searching assistance. They should also include disclosures of content sources and a privacy policy that details how personal data is handled. PMID- 15778133 TI - Yu Duan practices as embodying tradition, modernity and social change in Chiang Mai, Northern Thailand. AB - In this paper I examine the traditional postpartum beliefs and practices which still exist in northern Thailand today. The paper is based on qualitative research involving in-depth interviews with 30 women in Chiang Mai province. Beliefs and practices remain an essential part for postpartum care for women and have important consequences for women's health and wellbeing in northern Thailand. Many Thai women see their reproductive health problems as the consequence of inadequate postpartum practices. Thai women also believe that the effects of postpartum taboos would continue for the rest of their lives. Although the traditional postpartum beliefs and practices abound, the level of adherence differs according to the social structure of the women and their families. Poor rural women seem to hold on to their traditions more strongly than their urban counterparts. Urban middle class women in particular embody modernity in their thinking and behaviours concerning postpartum practices. But modernization has brought with it medical dominance. Due to their medical knowledge, doctors retain authority over both knowledge and status. The consequence of this dominance is the attempt to dismiss local traditional knowledge and practices. Although the pattern of traditional postpartum beliefs and practices is changing, it is still observed in northern Thailand. I contend that postpartum care for women incorporates local traditions so that women's health can be optimized at the time when they are in the most vulnerable stage of their lives. PMID- 15778134 TI - Gender-biased diagnosing of women's medical complaints:contributions of feminist thought, 1970-1995. AB - With the advent of second-wave feminism during the 1970s, a significant body of literature emerged describing sexist practices in women's health care. Gender biased diagnosing-the notion that somatic complaints by female medical patients are more likely to be labeled by physicians as psychosomatic-became a concern that garnered considerable attention in Europe and the United States because of the increased health risks it posed for women. This article examines the impact of feminist knowledge on this topic during the quarter century spanning 1970 1995. Analysis of the literature reveals feminist perspectives played a critical role in uncovering and problematizing gender bias in women's health care. PMID- 15778135 TI - Weight and behavioral and psychosocial factors among ethnically diverse, low income women after childbirth: I. Methods and context. AB - OBJECTIVE: In this paper, we present the background, hypotheses, methods, and descriptive findings from the Austin New Mothers Study, a longitudinal series measuring weight and contextual variables across the first postpartum year. Among the distinguishing features of this study are its tri-ethnic sample of low-income women and its serial measurement of the behavioral and psychosocial context of postpartum weight changes post-delivery and at 6 weeks, and 3, 6, and 12 months postpartum. METHOD: A prospective longitudinal design was used to assess body mass index (BMI), energy intake, fat intake, physical activity, health-related lifestyle, depressive symptoms, body image, and weight-related distress at each observation. SAMPLE: The analytic sample consisted of 382 White, African American, and Hispanic women. FINDINGS: For BMI, effects for ethnicity (p < .001), time (p < .001), and their interaction (p = .005) were significant. All ethnic groups had significant declines in BMI from delivery to 6 weeks postpartum. Thereafter, BMIs of White women declined during the second 6 months, whereas those of Hispanic and African American women either displayed small gains or plateaus between adjacent observations. Time-related effects were significant for behavioral and psychosocial variables except for body image and weight related distress, whereas effects for ethnicity were significant on fat intake, depressive symptoms, and body image. Despite declines in depressive symptoms, women remained at risk of depression across the first postpartum year. CONCLUSIONS: Low-income ethnic minority women have a higher vulnerability to postpartum weight gains or plateaus. High depressive symptoms occurred in all ethnic groups examined. PMID- 15778137 TI - Outcome evaluation of a multi-disciplinary community-based continence service for Australian women. AB - This longitudinal study evaluated the effectiveness of a multi-disciplinary community-based service offering conservative treatment for Australian women suffering urinary incontinence and living independently in the community, in terms of urinary incontinence symptom severity, impact on quality of life and knowledge outcomes. One hundred and twenty-three women attending The Waterworx Centre, a multi-disciplinary, publicly funded community-based continence service in South East Queensland Australia participated in the study. They received multi disciplinary conservative treatment for urinary incontinence, including comprehensive assessment and an individually-tailored plan of care. All the women were also linked back to their own generalist health professional for ongoing care and management. Data were collected over a one-year period: at first consultation, and at three months and six months following the first consultation. The International Continence Society Urinary Symptom Index Short Form-Female Outcome was used to measure urinary symptoms and impact on quality of life, and a researcher-developed test was used to measure changes in knowledge. Results showed that the women experienced an improvement in urinary symptoms and continence-related knowledge at three months following first consultation, and a decreased impact on quality of life, with these improvements either being sustained or increasing at six months. This study demonstrated that multi disciplinary community-based services offering specialist conservative treatment for women suffering urinary incontinence can be effective in achieving improvements in urinary symptoms and continence-related knowledge and reducing the impact of urinary incontinence on quality of life. PMID- 15778136 TI - Weight and behavioral and psychosocial factors among ethnically diverse, low income women after childbirth: II. Trends and correlates. AB - OBJECTIVE: This paper presents a longitudinal analysis of behavioral and psychosocial correlates of weight trends during the first postpartum year. Data are derived from the Austin New Mothers Study (ANMS), a longitudinal study of a low-income, tri-ethnic sample of postpartum women that incorporated serial assessment of weight and behavioral and psychosocial variables. METHOD: Postpartum body mass index (BMI) was measured prospectively (post-delivery, 6 weeks, and 3, 6, and 12 months postpartum). The analytic sample consisted of 382 White, African American, and Hispanic women receiving maternity care funded by Medicaid who had at least three measured postpartum weights. Behavioral and psychosocial variables included energy intakes, fat intakes, physical activity, health related lifestyle, smoking, breastfeeding, contraception, depressive symptoms, emotional eating, body image, and weight-related distress. RESULTS: Using hierarchical linear modeling to incorporate baseline only and time-varying effects, significant associations with postpartum BMI were found for the following variables: ethnicity (p = .001), time of weight measurement (p < .001), the interaction of ethnicity and time (p = .005), pre-pregnant BMI (p < .001), gestational weight gain (p < .001), weight-related distress (p < .001), and energy intakes (p = .005). After adjusting for covariates, ethnic groups displayed differing trends in postpartum BMI resulting in White women having significantly lower BMIs at 12 months postpartum compared to ethnic minority women (p's < .01). CONCLUSION: Behavioral and psychosocial variables contribute to a fuller understanding of BMI status of low-income women during the first postpartum year. PMID- 15778138 TI - Acute forensic medical procedures used following a sexual assault among treatment seeking women. AB - Despite the negative physical and mental health outcomes of sexual assault, a minority of sexually assaulted women seek immediate post-assault medical and legal services. This study identified the number and types of acute forensic medical procedures used by women presenting at a hospital-based urgent care centre between 1997 and 2001 within 72 hours following a reported sexual assault. The study also examined assault and non-assault factors associated with the use of procedures. It was hypothesized that assault characteristics resembling the stereotype of rape would be associated with the use of more procedures. The multiple regression indicated that injury severity, coercion severity, homelessness, and delay in presentation were significantly associated with the number of procedures received. Findings provide partial support for the hypothesis that post-assault procedures would be associated with the stereotype of rape, and highlight homeless women as a group particularly at risk for not receiving adequate medical treatment following a sexual assault. PMID- 15778139 TI - Development and implementation of a culturally sensitive cervical health survey: a community-based participatory approach. AB - Cervical cancer mortality rates are higher for Great Plains Native American women than for Caucasian women and other Native women. Messengers for Health, a project based on the Apsaalooke (Crow Indian) reservation, utilizes a lay health advisor approach to decrease cervical cancer screening barriers, increase knowledge regarding screening and prevention, and increase the proportion of women receiving Pap tests among Apsaalooke women aged 18 and older. This project utilizes a community-based participatory research model, which emphasizes community member involvement in all phases of the project. The initial phase of this project was the development and implementation of a culturally sensitive survey used to guide the program and benefit the community. The process and preliminary results are presented. PMID- 15778140 TI - Access to women's health care: a qualitative study of barriers perceived by homeless women. AB - Homelessness is an escalating national problem and women are disproportionately affected. Nevertheless, few studies have focused on the special circumstances associated with being a homeless woman. For instance, while both genders experience serious barriers to obtaining health care, homeless women face an additional burden by virtue of their sexual and reproductive health needs. The current study was conducted as the first stage of a qualitative/quantitative investigation of homeless women's access and barriers to family planning and women's health care. We interviewed 47 homeless women of diverse ages and ethnic backgrounds. A qualitative approach was initially taken to explore the factors homeless women themselves perceive as barriers to their use of birth control and women's health services, and factors they believe would facilitate their use. Key findings are that health is not a priority for homeless women, that transportation and scheduling can be particularly burdensome for homeless women, and that being homeless leads some to feel stigmatized by health care providers. Despite being homeless, having children was extremely important to the women in our study. At the same time, those interested in contraception confronted significant barriers in their efforts to prevent pregnancies. We conclude with suggested interventions that would make general, gynecological, and reproductive health care more accessible to homeless women. PMID- 15778141 TI - Perceptions of breast cancer among Arab Israeli women. AB - The incidence of breast cancer is increasing among Arab Israeli women, yet few Arab women over the age 50 take advantage of free mammography available to them. In this study we attempted to identify social and cultural aspects of the Arab Israeli woman's life that may reduce screening attendance for early detection of breast cancer. This qualitative study consisted of 6 focus groups. The fundamental theme identified as a barrier to prevention behavior was the role of the Arab woman in society expressed as marrying, having children and being totally committed to her family. In this model, without these roles her life is purposeless and her place in society is lost. The women perceived the chain of events that begins with screening for breast cancer as possibly resulting in cancer diagnosis and having breast cancer will lead to an inability to fulfill female roles. This feedback mechanism inherent in the chain of events may be the cause of the low rates of compliance in performing early detection and mammography. Interventions aimed at increasing the rate of Arab women's performance of mammography should take these issues into consideration. PMID- 15778142 TI - Positively influencing student nurse attitudes toward caring for elders: results of a curriculum assessment study. AB - Exposure to gerontological content has been shown to positively influence student attitudes toward elders. Our School of Nursing recently phased out one curriculum while implementing a new integrated curriculum. In the former, aging content was presented in a first-semester senior course. In the new curriculum, students were introduced to aging concepts in first-semester junior courses and additional content was integrated throughout the program. The purpose of this study was to evaluate the effectiveness of the new integrated curriculum compared to the former in positively influencing attitudes toward caring for elders. An attitudes questionnaire was administered at 3 time points to students in each curriculum. Both groups, but particularly the integrated group, demonstrated statistically significant improvements in attitudes over time. PMID- 15778143 TI - Integrating geriatric content into a medical school curriculum: description of a successful model. AB - Most medical school curricula do not equip students with adequate attitudes, knowledge and skills to care for elderly populations. We describe an effective geriatric curricular infusion model compatible with preserving the overall curricula schema. Course and clerkship directors, staff and faculty from the Office of Educational Development, Center on Aging, curriculum committee and Associate/Assistant Deans of Education, and faculty from the schools of medicine, nursing, and allied health collaborated in the effort. Each of these components and institutional financial commitment were critical to successful basic science and clinical geriatric content infusion addressing the American Geriatric Society (AGS) Core Competencies. Delivery modalities included problem- based learning cases, lectures, standardized patient portrayals for teaching and assessment, and experiential activities with elderly. Assessments were conducted and outcomes tracked in several ways, including: (1) annual course reviews, focus groups, and student evaluations; (2) mandatory geriatrics 4th year graduation competency exam; and, (3) AAMC Graduation Questionnaire responses. Initial data indicate that student knowledge and competencies have increased with increasing exposure in the desired areas, and support infusion as a viable approach to enhancing gerontology and geriatric curricular content. PMID- 15778144 TI - Geriatric medical education in Israel. AB - In this article we will focus on geriatric medical education in Israel and will review our experience in this field. A coordinated effort of the Ministry of Health and the Israeli Medical Association led to the establishment of a modern geriatric system and to the recognition of geriatrics as a medical specialty in the early 1980s. All four Israeli medical schools joined this effort by introducing geriatrics in the undergraduate studies. Several innovative models for teaching geriatrics to medical students and for postgraduate training have been developed and implemented. The growing body of skills and knowledge available, and their application for competent care of the elderly, constitute a compelling impetus for the further development of a high-quality geriatric medical education. PMID- 15778145 TI - Web-based framework for improving geriatric education. AB - Despite the growth in the elderly population, physicians with special geriatric training and certification number only 9,000 out of 650,000 doctors in the United States. The flexibility and increasing availability of the Internet makes it an ideal avenue for addressing the educational needs of health care providers to improve the health and care of older adults. We performed an Internet search using popular search engines. This yielded between 321 and 358,000 websites about geriatrics and 164 to 175,000 about geriatric education. This paper reviews popular health and medical Web sites to determine geriatric educational content and proposes a structure for an "ideal" geriatric Web site that would provide resources and enhance training for allied health professionals, physicians in training and community doctors in practice. We also propose a framework addressing the design and content for developing a geriatric specific website. We use this framework to propose a prototype resource on falls and gait instability. PMID- 15778146 TI - Nursing home staff characteristics and knowledge gain from a didactic workshop on depression and behavior management. AB - Depression is a prevalent and serious problem among nursing home residents. Nursing home staff members are gatekeepers for mental health treatment for residents, but may know little about depression and its management. We evaluated a didactic workshop for nursing home staff on depressive symptoms and management. Results for 58 staff participants showed significant gains in knowledge about depression diagnosis and management after the workshop. A regression predicting pre-test knowledge indicated that significant contributors were age (beta = .27; p < .05), education (beta = .28, p = < .05), and experience in long-term care (beta = -.53, p < .01). Results provide preliminary support for didactic training about depression, but suggest that differences in staff knowledge related to experience should be addressed in training. PMID- 15778147 TI - A pilot study of CME on risk management in long-term care. AB - This pilot study's purpose was to evaluate behavioral changes among medical directors and physicians following CME on risk management in long-term care (LTC) facilities. The setting was a satellite conference at the AGS Meeting Symposium 2000. CME participants included 51 medical directors, attending physicians, and nurses. Evaluations were based on 20 LTC medical directors and physicians. Descriptive statistics were used for outcome measurements. Respondents (n = 15, 75%) "committed" to an average of 3 changes. Of the 45 commitments, no progress was reported on 8 (17.7%); another 8 commitments were reported as fully completed. The mean implementation rate was 60%. LTC physicians departed the course with intentions to alter their behaviors, but important obstacles such as "lack of time" and "staff not available or interested" interfered with implementation. PMID- 15778149 TI - A survey of evaluation practices for hospice social workers. AB - Hospice social workers make distinct contributions to the care of terminally ill persons and their loved ones. This study contributes to social work's professional knowledge base by examining methods by which hospice social workers are evaluated by their supervisors as well as their clients. This article reports on a survey of 109 hospices' evaluative methods for social workers. Implications for patient care and hospice operations are discussed. PMID- 15778150 TI - Penny wise, pound wise: a comparison of Medicaid expenditures for home and community-based services versus nursing facility care for older adults. AB - Kansas Medicaid offers home and community-based services through the Frail and Elderly (FE) program as an alternative for older adults who are eligible for nursing home (NH) care but wish to stay in the community. We determined demographic and clinical characteristics of enrollees receiving FE or NH services, examined their relative health care utilization patterns, and estimated the difference in Medicaid's expenditures for FE versus NH care. After adjusting for key demographic and clinical variables, mean monthly expenditures were $1,281 lower for the FE cohort. Since NH and FE populations are dissimilar, these care options may not be easily interchangeable at the individual level. PMID- 15778151 TI - Living arrangements decisions at discharge and later: differences in criteria and outcomes. AB - This study examined the pattern of post rehabilitation living arrangements over 18 months of 172 adults discharged from a geriatric rehabilitation unit at a large urban medical center, all of whom were living alone prior to admission. Results showed the diminishing importance of physical function and cognition as factors in decisions to return home as the length of time after hospital discharge increased. Patients who returned home to live alone after a period of time living in more supported environments had significantly lower assessment scores on physical function (p < 0.001) and cognition (p < 0.001) compared to patients who went home to live alone immediately upon discharge. As well, patients who returned home to live alone later than three months post discharge were significantly more likely to move to more supported environments by 18 months after discharge (p = 0.043) and to experience re-hospitalizations (p = 0.008), which raises questions about the appropriateness of these later decisions. PMID- 15778152 TI - Organizational and environmental factors affecting the utilization of telemedicine in rural home healthcare. AB - In the last five years, home health agencies have become increasingly interested in telemedicine as a potential means to meet the future healthcare needs of their aged and chronically ill clientele. This case study examines the organizational and environmental conditions that affected the implementation of a telemedicine program in one rural home healthcare organization. Several factors restricted the utilization of telemedicine, including Medicare's Prospective Payment System and corresponding documentation (Outcome Assessment and Information Set), the organization controlling grant funding for the program, and several environmental factors. Findings suggest that in rural communities, older homecare patients may have less opportunity to benefit from telemedicine. The study demonstrates the importance of environmental and organizational factors when implementing a telemedicine program. Recommendations are offered for home healthcare organizations considering development of telemedicine programs. PMID- 15778154 TI - A questionnaire to examine food service satisfaction of elderly residents in long term care facilities. AB - The purpose of this study was to develop a survey tool for assessing the satisfaction of elderly long-term care (LTC) residents with the meals and food services they receive, as well as to assess quality of life issues related to eating. Food service delivery should be provided in an environment that fosters autonomy, interpersonal relations, and security. The questionnaire was administered as face-to-face interviews with 205 residents (> or = 65 years of age) of 13 LTC facilities in Saskatoon, Saskatchewan, Canada (participation rate = 67%). Residents expressed some concern with food variety, quality, taste, and appearance, and with the posting of menus. Quality of life issues were mostly positive; however, residents were less satisfied with areas related to their autonomy such as food choice and snack availability. PMID- 15778155 TI - What older adults find useful for maintaining healthy eating and exercise habits. AB - Four focus groups were conducted with a total of 29 adults 60 years of age and older enrolled in the SENIOR Project, a health promotion intervention study designed to increase fruit and vegetable consumption and exercise among community dwelling older adults. The focus groups explored the motivations of older adults to eat five or more servings of fruits and vegetables and/or exercise and the strategies used to adopt or maintain these behaviors. Participants stated that maintaining health, remaining independent, and fearing illness provided the motivation needed to adhere to these behaviors. The strategies or the behavioral processes used to adopt or maintain these behaviors included counterconditioning, helping relationships, stimulus control, and self-liberation. PMID- 15778156 TI - Nutrition risk and associated factors in congregate meal participants in northern Florida: role of Elder Care Services (ECS). AB - Nutrition is a key for extending quality of life of the elderly. Factors associated with nutritional risk in elders were identified with a modified nutrition screen initiative checklist. High, moderate and low nutritional risk was seen in 31, 46 and 23%, respectively, of 324 elders in six congregate meal sites in northern Florida. High nutritional risk was associated with urban residency, females, being African American, self-reported poor health and irregular visits to medical/health professionals. African American females living in urban areas with limited access to health care are most vulnerable. PMID- 15778157 TI - Body composition and nutrient intake of hypertensive elders and middle-aged men and women in Nigeria. AB - Body composition of eighty hypertensive patients (43 middle-aged, 30-59 years and 37 elders, 60-80 years) was determined by anthropometry alongside their dietary intake. The mean body weight and height of female elders were 54.1 +/- 14.6 kg and 157.0 +/- 7.0 cm while that of male elders were 61.8 +/- 13.5 kg and 167.1 +/ 9.0 cm. Corresponding values for middle-aged subjects were also determined. The middle-aged subjects were heavier and taller than the elders in both sexes. Among the elders BMI values 21.7 (females) and 22.1 (males) were normal, but the female had higher percentage body fat (28.0 +/- 0.1%) than males (20.8 +/- 5.2%). Other body composition parameters determined included lean body mass, optimum body weight, body water, skin-fold thickness and arm muscle diameter. BMI was significantly correlated with high blood pressure. The mean energy and nutrient intake of the subjects were inadequate except for the female elders. PMID- 15778159 TI - Needs and preference assessment for an in-home nutrition education program using social marketing theory. AB - Nutrition education programs for elder caregivers (CG) and their elder care recipients (CR) are important in preventing malnutrition. Using Social Marketing Theory, this study assessed the needs and preferences for nutrition education in elder CGs and their CRs in Guilford County, NC. Thirty-two pairs of community residing elder CGs/CRs and three focus groups (FGs) participated. Health and diet questionnaires were administered to all CGs/CRs during in-home interviews. CGs/CRs and FGs evaluated nutrition education materials. Questionnaires were analyzed using SPSS v9. Ethnograph v5.0 was used to code the interviews regarding the education materials. The CGs were middle age (58.9 years), overweight (BMI = 28.8) Caucasian women. The CRs were old (79.4 years), overweight (BMI = 26.0) Caucasian women. Identified malnutrition risk factors of CGs and CRs included inadequate fluid and dietary intake, polypharmacy, and chronic disease. Identified nutrition needs and education preferences of CGs/CRs were similar. Perceived nutrition education preferences of the FGs did not reflect the interests of the CGs/CRs. This information is being used to revise the education materials and develop an in-home nutrition education program for CGs and CRs in Guilford County, NC. PMID- 15778166 TI - Alone and content: frail seniors living in their own homes compared to those who live with others. AB - This study compared the psychosocial well-being and functioning of frail elderly people who live alone to those who live with others. An exploratory study was conducted with a sample of 5,265 members of a social HMO in Southern California. Those living alone were more likely to be widowed, female, and European American. Elderly people living with others reported higher levels of depression and poorer functioning. Both groups reported having a social support network, but the sources of support varied. This study contradicts older research with its finding that living alone is not synonymous with depression. Further research is recommended. PMID- 15778167 TI - Understanding and reducing stress and psychological distress in older women with heart disease. AB - Stress and psychological distress were assessed in 457 older women who were subsequently randomized to a six-week heart disease management program ("Women take PRIDE") or to a "usual care" control group. Baseline distress was significantly associated with age, symptoms, physical functioning, social support, optimism, and self-esteem (p < .05). Only 20% of women reported their physician had recommended reducing stress. At four months follow-up, intervention women compared to controls were significantly more likely to report reductions in stress levels (p = 0.02) and also showed improvement in emotional behavior (p = 0.09). PMID- 15778168 TI - Women's financial planning for retirement: the impact of disruptive life events. AB - Providing care for an aged parent has immediate financial, emotional, psychological, and physical consequences for the primary caregiver. This pilot study of 138 middle aged and older females analyzes the long term financial consequences of providing care to aged relatives for female caregivers. The impact of this disruptive life event (e.g., caring for an aged relative) on retirement planning among middle aged and older adult women was analyzed using quantitative data collected from women residing in the Midwest region of the United States. PMID- 15778169 TI - Gender differences in factors that influence time spent planning for retirement. AB - Retirement research has shown that planning activities are influenced by a variety of demographic and psychological variables. However, few investigations have focused on how demographic and psychological factors influence the time and effort men and women allocate to retirement preparation. In the present study, 184 individuals completed a survey designed to assess future time perspective (FTP), worry about retirement, age, and income level. Hierarchical regression techniques were used to examine how these variables influenced the amount of time men and women spent planning for and thinking about retirement. Results revealed that women spent less time thinking about retirement than men, and gender was differentially related to the factors predictive of this activity. From an applied perspective, these findings suggest that separate retirement intervention programs are warranted that meet the unique needs of working men and women. PMID- 15778170 TI - Gender difference in physical performance, body composition and dietary intake in homebound elders. AB - Considering that the most vulnerable subgroup of elders for poor nutritional status and functional decline is the increasing number of homebound women, we examined gender differences in physical performance, body composition, and dietary intake in a randomly recruited sample of 345 homebound elders (81% women, 48% black, > 65% income < $750/month). After controlling for demographic and health-related factors, the results from multivariate analyses indicated that women were more likely than men to report the lowest nutrient intake and were 2.9 times more likely to be at the worst level of overall physical performance. These results suggest a heightened vulnerability of homebound older women to poor physical performance, low dietary intake, and increased Body Mass Index (BMI). Prospective research is now needed to examine the interrelationships between physical performance, dietary intake, and body composition among the growing homebound older population. PMID- 15778171 TI - Risk factors associated with osteoporosis in Hispanic women. AB - This study examined risk factors for osteoporosis in Hispanic women. Factors examined included ethnicity, gender, age, height, weight, family and personal history of fractures, height loss, exercise, diet, time since menopause or hysterectomy, hormone replacement therapy (HRT), calcium supplementation, hypertension, thyroid disease, diabetes, arthritis, chemotherapy, family history of breast cancer, use of water pills, fosamax, steroids, alcohol, and smoking. Most results found parallel those found in the Caucasian population. Heavier patients had greater bone density, as well as patients who exercised and those using HRT. Older patients had lower bone density as did diabetic patients. Results not anticipated were higher bone density in patients not taking calcium supplements, and in patients who consumed alcohol. PMID- 15778172 TI - Older women on the U.S.-Mexico border: exploring the health of Hispanics and non Hispanics Whites. AB - Research on ethnicity and socioeconomic status (SES) suggests that Hispanics are more likely than non-Hispanic Whites to experience poverty and low levels of education, which may relate to poorer health status. This study used a health survey to examine income, education, ethnicity, birthplace, and age on self reported health factors of women age 60 and older on the U.S.-Mexico border. Results show that income, age, and education were significantly associated with several health factors (Physical Health, Emotional Health, General Health, Energy Level, and Activity Potential). Older women with lower SES, regardless of ethnicity, reported poorer health than younger-old women with higher SES. PMID- 15778173 TI - Persevering: the experience of well elderly women overcoming the barriers to the U.S. health care system. AB - PURPOSE: To explore the barriers to health care for well older community living women living in a metropolitan area in North Central Texas. METHODS: Qualitative thematic design. Forty-six women who attended seniors' centers in North Central Texas participated in focus groups. Then three individual and a key informant interviews were completed. Data were analyzed using Lubrosky's thematic analysis. FINDINGS: The participants described barriers to access associated with provider barriers of devaluing older adults, the complexity and slowness of reimbursement, inadequacy of health insurance plans and physicians opting out of plans. Participants overcame these barriers by persevering. PMID- 15778174 TI - Pioneers in our own lives: grounded theory of lesbians' midlife development. AB - Historically, human development theorists made no mention of experiences unique to lesbians. To explore implications of differences associated with lesbian midlife development, this qualitative study compared the midlife experiences of 12 lesbians between the ages of 41 and 54 to descriptions of midlife development resulting from two prior studies using similar methods with groups whose members were nearly all heterosexual women, i.e., data collected in a series of focus group meetings and analyzed using a grounded theory approach. While lesbians in this study faced many of the same occurrences described by other middle-aged women, they experienced those events in different ways. The lesbians experienced less emotional turmoil during midlife than did their heterosexual counterparts. They also expressed unique concerns about old age. PMID- 15778175 TI - Women's adjustment to widowhood: does social support matter? AB - Widowhood is a stressful event for women, often coinciding with health, financial, and relationship losses. Researchers have considered many aspects of women's adjustment to widowhood, typically including social support in analyses. This study analyzed relationships between social support and adjustment from 19 previous widowhood studies. Contrary to intuition and most researchers' assumptions, the majority of relationships did not show that social support positively impacted adjustment. Analyses examined whether type of support or study methods affected the likelihood of finding a significant relationship between support and adjustment. Chi-square statistics indicated that neither of these factors influenced the effect of social support on women's adjustment to widowhood. PMID- 15778176 TI - Biographical sketches of elderly African-American women in later life. AB - The research concerning the African-American elderly is limited, since this group is made up of less than eight percent of the entire U.S. population. Due to discriminatory practices in the U.S., based on race and gender, the African American elderly woman has some unique issues facing her in old age. This paper will present three qualitative interviews of African-American women from age 65 to 100. These biographical sketches will report how these women dealt with family traditions, lifestyle changes, and growing old in their lives. Their perceptions of personal joys and frustrations, plus attitudes about how race, not gender, was a significant factor in their lives, will be reported. Education and religion were life-long strengths in their lifestyle. PMID- 15778178 TI - PubMed enhancements: fulfilling the promise of a great product. AB - There have been many recent changes to PubMed to enhance its usefulness. Those changes include: LinkOut Libraries (local holding field), PubMed Central (full text articles archived by the National Library of Medicine), and LinkOut (access to full-text articles right from the PubMed citation). Medical librarians should be aware of how these features work to best assist their clients. These new features offer the possibility of true desktop access for library patrons. Not only will patrons appreciate these new features, but their use in libraries will literally change what we do, who does it, and how it is done. PMID- 15778179 TI - Family practice clerkship information seminars:the role of the health sciences library. AB - This article will describe the New Jersey Medical School (NJMS) Family Medicine clerkship's Information Seminars for thirdyear medical students, conducted by the faculty and staff of the Department of Family Medicine and by librarians at the University of Medicine and Dentistry of New Jersey's (UMDNJ) George F. Smith Library of the Health Sciences. Content of the information seminars will be briefly described, as well as the renewed focus on the library through the incorporation of evidence-based medicine concepts into practice. PMID- 15778180 TI - Educating generation X and generation Y: teaching tips for librarians. AB - This article provides a list of helpful teaching tips for instructional librarians who need to meet the changing generational needs of their patrons. Specific generational qualities and attitudes of Generation X and Generation Y are discussed along with educational techniques and software recommendations. These tips are based on the authors' experiences at Drexel University's Hahnemann Library and Thomas Jefferson University's Scott Memorial Library, both of which are academic health sciences libraries. PMID- 15778181 TI - Search strategies for population and social subjects in a medical school curriculum. AB - This case study describes strategies developed for medical students to conduct Specific (as opposed to Sensitive) searches in PubMed so they can quickly locate several relevant references to articles on the population and social aspects of medicine. The Specific search strategy protocol for population and social aspects of medicine involves use of the MeSH Database, certain features of the Detailed Display in the MeSH Database, and applying Limits. By teaching the Specific form of search strategy, busy users can learn a handful of basic techniques that should yield a useful proportion of references, thereby ensuring these users' frequent success. PMID- 15778182 TI - Are we safe? NLM's household products database. AB - This column features an overview of the Division of Specialized Information Services, National Library of Medicine Household Products Database. Basic searching techniques are presented, as well as a brief overview of the data contained in this file. The Household Products Database contains information on chemical ingredients in various products used in U.S. homes. PMID- 15778183 TI - Addressing health disparities: African American consumer information resources on the web. AB - The Web continues to rapidly evolve as a medium of information exchange. One major area where the Web is now playing a definitive role is in increasing consumer accessibility to health information for personal health and disease related questions. The comprehensiveness of health information on the Web has the potential to empower consumers when making critical health-related decisions. Conversely, the myriad of information resources presents an array of challenges when seeking specific, credible, and timely health information. As the United States health care system continues to change, community concerns about the quantity, quality, and broad nature of information has become critical. In particular, the Web can play a vital role in health awareness and promotion by disseminating health information for traditionally underrepresented groups, particularly African Americans. Increased accessibility for this group has not always been the focus of attention; however, the growth of the Web coupled with heightened awareness of public health initiatives brings to the forefront numerous opportunities to provide tools that promote essential health information literacy. PMID- 15778184 TI - What did they know? And when did they know it? Progress in helping clinicians reach decisions. AB - In the past three decades, knowledge professionals have made remarkable strides toward achieving clinicians' decision-support dream of immediately available short answers. Medical evidence, what is truly known about health care effects and ready to be applied in patient care, can now much more easily be identified. Today's information delivery tools enable clinicians to seek answers to many more of their questions and to find the answers when patient encounters and teachable moments are occurring. Excerpted from a state health sciences library association presentation, this paper highlights efforts to speed clinician adoption of health care discoveries into routine patient care. PMID- 15778185 TI - Getting on the fast track, or how to get an MLIS through distance education, with a specialization in medical librarianship. AB - The article describes an innovation in MLIS education for medical librarianship, with an introduction to the FastTrack, the distance education program at the University of Pittsburgh's library and information science school, together with an overview of a model program linking the biomedical library at Vanderbilt University with the School of Information Sciences in Pittsburgh. Admissions requirements and specific curriculum for the distance education master's degree are detailed in an FAQ format. PMID- 15778202 TI - Combining Parenting and a Science Career. PMID- 15778203 TI - Crying "Whorf" PMID- 15778204 TI - Prediction of sudden cardiac death after acute myocardial infarction: role of Holter monitoring in the modern treatment era. AB - AIMS: Current treatment may have changed the risk profiles of survivors of acute myocardial infarction (AMI). We evaluated the utility of Holter-based risk variables in the prediction of sudden cardiac death (SCD) among survivors of AMI treated with modern therapy. METHODS AND RESULTS: A total of 2130 AMI patients (mean age 59 +/- 10 years) were included. The patients were treated with modern therapeutic strategies, for example, 94% were on beta-blocking therapy and 70% underwent coronary revascularization. Various risk parameters from Holter monitoring were analysed. During a median follow-up of 1012 days (interquartile range: 750-1416 days), cardiac mortality was 113/2130, including 52 SCDs. All Holter variables predicted the occurrence of SCD (P<0.01), but only reduced post ectopic turbulence slope (TS) (P<0.001) and non-sustained ventricular tachycardia (P<0.01) remained as marked SCD predictors after adjustment for age, diabetes, and ejection fraction (EF). In a subgroup analysis, none of the Holter variables predicted SCD among those with an EF < or = 0.35, but many variables predicted SCD among those with an EF >0.35, particularly TS (hazard ratio 5.9; 95% CI 2.9 11.7, P<0.001). CONCLUSION: Among the post-AMI patients treated according to the current guidelines, the incidence of SCD is low. Various Holter variables still predict the occurrence of SCD, particularly among the patients with preserved left ventricular function. PMID- 15778205 TI - The clinical characteristics and investigations planned in patients with stable angina presenting to cardiologists in Europe: from the Euro Heart Survey of Stable Angina. AB - AIMS: The Euro Heart Survey of Stable Angina set out to prospectively study the presentation and management of patients with stable angina as first seen by a cardiologist in Europe, with particular reference to adherence to existing guidelines and regional variability in patient presentation and initial assessment. METHODS AND RESULTS: Consecutive outpatients with a clinical diagnosis by a cardiologist of stable angina were enrolled in the study and 3779 patients were included in the analysis. The average age was 61 years and 58% were male. The majority of patients (88%) had mild to moderate angina, CCS class I or II. Despite a high prevalence of recognized risk factors, 27% did not have cholesterol and 33% did not have glucose measured within 4 weeks of assessment. The resting ECG was abnormal in 41% of patients. An exercise ECG was performed or planned as part of initial investigation in 76% of patients and 18% had a stress imaging test such as perfusion scanning or stress echo. A coronary angiogram was performed or planned in 41%, and 64% had an echo. The time from assessment to investigation varied widely, particularly for angiography. One in 10 patients had neither any form of stress test nor angiography, with marked regional variation. Availability of invasive facilities increased the likelihood of both non-invasive and invasive investigations. Those with more severe symptoms or longer duration of symptoms or a positive non-invasive test were more likely to have angiography. In multivariable analysis, a positive stress test was the strongest predictor of the use of angiography, associated with a six-fold increase in the likelihood of invasive investigation. However, gender and availability of facilities were also predictive. CONCLUSION: Considerable variation in features at presentation and use of investigations has been identified in the stable angina population in Europe. The evaluation of biochemical cardiovascular risk factors was suboptimal. Overall rates of non-invasive investigation for angina and the clinical appropriateness of factors predictive of the use of invasive investigation were broadly in line with guidelines. However, the influence of access to facilities, and marked international variation in rates and timing of investigation suggest that factors unrelated to clinical need are also influential in the management of patients with stable angina. PMID- 15778207 TI - Ethics and deafness: a matter of perspective? PMID- 15778208 TI - The question of sign-language and the utility of signs in the instruction of the deaf: two papers by Alexander Graham Bell (1898). AB - Alexander Graham Bell is often portrayed as either hero or villain of deaf individuals and the Deaf community. His writings, however, indicate that he was neither, and was not as clearly definite in his beliefs about language as is often supposed. The following two articles, reprinted from The Educator (1898), Vol. V, pp. 3-4 and pp. 38-44, capture Bell's thinking about sign language and its use in the classroom. Contrary to frequent claims, Bell does not demand "oral" training for all deaf children--even if he thinks it is the superior alternative--but does advocate for it for "the semi-deaf" and "the semi-mute." "In regard to the others," he writes, "I am not so sure." Although he clearly voices his support for oral methods and fingerspelling (the Rochester method) over sign language, Bell acknowledges the use and utility of signing in a carefully-crafted discussion that includes both linguistics and educational philosophy. In separating the language used at home from that in school and on the playground, Bell reveals a far more complex view of language learning by deaf children than he is often granted. (M. Marschark). PMID- 15778209 TI - Phonological processing in deaf children: when lipreading and cues are incongruent. AB - Deaf children exposed to Cued Speech (CS), either before age two (early) or later at school (late), were presented with pseudowords with and without CS. The main goal was to establish the way in which lipreading and CS combine to produce unitary percepts, similar to audiovisual integration in speech perception, when participants are presented with synchronized but different lipreading and auditory information (the McGurk paradigm). In the present experiment, lips and cues were sometimes congruent and sometimes incongruent. It was expected that incongruent cues would force the perceptual system to adopt solutions according to the weight attributed to different sources of phonological information. With congruent cues, performance improved, with improvements greater in the early than the late group. With incongruent cues, performance decreased relative to lipreading only, indicating that cues were not ignored, and it was observed that the effect of incongruent cues increased when the visibility of the target phoneme decreased. The results are compatible with the notion that the perceptual system integrates cues and lipreading according to principles similar to those evoked to explain audiovisual integration. PMID- 15778210 TI - Parents' of deaf children evaluative accounts of the process and practice of universal newborn hearing screening. AB - This article presents results from a narrative interview study of 45 parents/caregivers whose infants were correctly identified as deaf through Phase 1 of the Newborn Hearing Screening Programme in England. It concerns the period from the first screening event to the point of referral for audiological assessment. It focuses on the meanings parents attribute to the inconclusive message that the screen delivers and analyzes what it is that differentiates parents for whom such an outcome raises little concern from those who express dissatisfaction. Parents' evaluations of specific features of screening practice and process such as communication style and manner are also considered. It ends with a discussion of the status and validity of parents' accounts within the context of an evaluation of a national screening program and the further development of professional practice. PMID- 15778211 TI - Predicting career development in hard-of-hearing adolescents in Australia. AB - This article reports on a study investigating the career development of hard-of hearing high school students attending regular classes with itinerant teacher support. We compared 65 hard-of-hearing students with a matched group of normally hearing peers on measures of career maturity, career indecision, perceived career barriers, and three variables associated with social cognitive career theory career decision-making self-efficacy, outcome expectations, and goals. In addition, the predictors of career maturity and career indecision were tested in both groups. Results indicated that (a) the two groups did not differ on measures of career maturity, (b) the SCCT variables were less predictive of career behaviors for the hard-of-hearing students than for the normally hearing students, and (c) perceived career barriers related to hearing loss predicted lower scores on career maturity attitude for the hard-of-hearing students. These findings are discussed in the context of career education and counseling interventions that may benefit young people who are hard of hearing. PMID- 15778212 TI - Initiations of social interactions by young hearing impaired preschoolers. AB - This study examined strategies for initiating social interactions with peers, among 4 children with hearing impairment, aged 33 to 36 months, attending a special early education center or a regular kindergarten. The study investigated initiation type (related to partner's hearing status) and rates of initiation success/failure vis-a-vis hearing and deaf partners. Results revealed (a) more initiations in the regular program than in the special program; (b) in the special program, much more successful initiations toward children with hearing impairment than toward hearing children; (c) vocalization as the most frequent strategy used with both hearing and hearing-impaired partners; and (d) referential decisions about their initiations even among young children with hearing impairment (made by changing frequencies of various strategies according to partner's hearing status). The discussion addressed implications regarding integration of children with hearing impairment into regular educational settings. PMID- 15778213 TI - Ethical reasoning and mental health services with deaf clients. AB - Ethical problems encountered by mental health practitioners working with deaf clients are often complex and involve issues not fully addressed in professional codes of ethics. A principles-based ethical reasoning process can assist in resolving many of these ethical concerns. Principles such as beneficence, nonmaleficence, autonomy, fairness, integrity, and respect are found in the ethical codes of many disciplines; these can also create a common language or reference point when professionals from different fields attempt to deal with shared problems. This article discusses some applications of these principles in working with deaf individuals and proposes an ethical decision-making process that can provide a framework for ethical reasoning in thinking through complex problems. PMID- 15778214 TI - Ethics, deafness, and new medical technologies. AB - In the last 50 years, several new technologies have become enormously important within the Deaf community and have helped significantly to improve deaf people's lives in a hearing world. Current public attention and admiration, however, seems unduly focused on medical technologies that promise to solve "the problem" of being deaf. One reason for this interest, we argue, is the public's preoccupation with deafness as a disability and promises that technological breakthroughs such as the cochlear implant will "cure" deafness. Pressure on parents to make quick and early decisions and lack of adequate information about alternatives often leave them unprepared for the consequences of these decisions. To allow deaf individuals and their families to make better informed decisions about their lives and their futures, we argue finally that professionals who interact with these families adopt inclusive and individualizing ethics. PMID- 15778215 TI - An unhappy and utterly pitiable creature? Life and self-images of deaf people in the Netherlands at the time of the founding fathers of deaf education. AB - This article describes how young deaf people in the Netherlands between 1809 and 1828 made the transition from living in a school for the Deaf,1 a rather protected community with mostly deaf people and with hearing people who could understand them rather well, to a life in hearing society with mostly hearing people who knew little about deafness. How did they manage to live in that hearing society? The article describes how these deaf people viewed themselves as Deaf persons in a hearing society. The description is based on an analysis of 73 letters written by 35 ex-pupils to the founder of their school, Reverend H. D. Guyot. As it turns out, these deaf ex-pupils managed to live in hearing society remarkably well. PMID- 15778216 TI - Evaluation of the support services provided to deaf children attending secondary general schools in Cyprus. AB - Deaf children have been integrated into secondary schools in Cyprus since 1990. This article reports the results of a major study carried out in Cyprus, the aim of which was to evaluate for the first time the support services available for deaf children receiving their education in secondary general schools. For the purposes of our study, four types of questionnaires were designed and administered to all deaf children integrated into secondary general schools, as well as to their parents, teachers, and head teachers. All participants stressed that the support services are vital for the children's academic success and social integration. The main support services identified by the participants in our study were: one-to-one and group sessions; presence of co-coordinators (special teachers of the deaf); "acoustical treatment" of the classrooms; provision and management of personal amplification, psychological support, and counseling; and in-service training for designated teachers. The majority of the participants expressed satisfaction with the support services offered to them. They also made some suggestions for the improvement of the integration of deaf children into secondary schools in Cyprus. PMID- 15778217 TI - Arabic sign language: a perspective. AB - Sign language in the Arab World has been recently recognized and documented. Many efforts have been made to establish the sign language used in individual countries, including Jordan, Egypt, Libya, and the Gulf States, by trying to standardize the language and spread it among members of the Deaf community and those concerned. Such efforts produced many sign languages, almost as many as Arabic-speaking countries, yet with the same sign alphabets. This article gives a tentative account of some sign languages in Arabic through reference to their possible evolution, which is believed to be affected by the diglossic situation in Arabic, and by comparing some aspects of certain sign languages (Jordanian, Palestinian, Egyptian, Kuwaiti, and Libyan) for which issues such as primes, configuration, and movement in addition to other linguistic features are discussed. A contrastive account that depicts the principal differences among Arabic sign languages in general and the spoken language is given. PMID- 15778218 TI - Biochemical properties of Saccharomyces cerevisiae DNA polymerase IV. AB - Although mammals encode multiple family X DNA polymerases implicated in DNA repair, Saccharomyces cerevisiae has only one, DNA polymerase IV (pol IV). To better understand the repair functions of pol IV, here we characterize its biochemical properties. Like mammalian pol beta and pol lambda, but not pol mu, pol IV has intrinsic 5'-2-deoxyribose-5-phosphate lyase activity. Pol IV has low processivity and can fill short gaps in DNA. Unlike the case with pol beta and pol lambda, the gap-filling activity of pol IV is not enhanced by a 5'-phosphate on the downstream primer but is stimulated by a 5'-terminal synthetic abasic site. Pol IV incorporates rNTPs into DNA with an unusually high efficiency relative to dNTPs, a property in common with pol mu but not pol beta or pol lambda. Finally, pol IV is highly inaccurate, with an unusual error specificity indicating the ability to extend primer termini with limited homology. These properties are consistent with a possible role for pol IV in base excision repair and with its known role in non-homologous end joining of double strand breaks, perhaps including those with damaged ends. PMID- 15778219 TI - Up-regulation of human myo-inositol oxygenase by hyperosmotic stress in renal proximal tubular epithelial cells. AB - myo-Inositol oxygenase (MIOX) catalyzes the oxidative cleavage of myo-inositol (MI) to give d-glucuronic acid, a committed step in MI catabolism. d-Glucuronic acid is further metabolized to xylitol via the glucuronate-xylulose pathway. Although accumulation of polyols such as xylitol and sorbitol is associated with MI depletion in diabetic complications, no causal relationship has been established. Therefore we are examining the role of MIOX in diabetic nephropathy. Here we present evidence that the basis for the depletion of MI in diabetes is likely to be mediated by the increased expression of MIOX, which is induced by sorbitol, mannitol, and xylitol in a porcine renal proximal tubular epithelial cell line, LLC-PK1. To understand the molecular mechanism of regulation of MIOX expression by polyols, we have cloned the human MIOX gene locus of 10 kb containing 5.6 kb of the 5' upstream sequence. Analysis of the 5' upstream sequence led to the identification of an osmotic response element (ORE) in the promoter region, which is present approximately 2 kb upstream of the translation start site. Based on luciferase reporter and electrophoretic mobility shift assays, polyols increased the ORE-dependent expression of MIOX. In addition, we demonstrate that the activity of the promoter is dependent on the binding of the transcription factor, tonicity element-binding protein, or osmotic response element-binding protein, to the ORE site. These results suggest that the expression of MIOX is up-regulated by a positive feedback mechanism where xylitol, one of the products of MI catabolism via the glucuronate-xylulose pathway, induces an overexpression of MIOX. PMID- 15778220 TI - Dynamic CpG and non-CpG methylation of the Peg1/Mest gene in the mouse oocyte and preimplantation embryo. AB - In somatic tissues, the CpG island of the imprinted Peg1/Mest gene is methylated on the maternal allele. We have examined the methylation of CpG and non-CpG sites of this differentially methylated CpG island in freshly ovulated oocytes, in vitro aged oocytes, and preimplantation embryos. The CpG methylation pattern was heterogeneous in freshly ovulated oocytes, despite the fact that they all were arrested in metaphase II. After short in vitro culture, Peg1/Mest became hypermethylated, whereas prolonged in vitro culture resulted in demethylation in a fraction of oocytes. Non-CpG methylation also occurred in a stage-specific manner. On alleles that were fully methylated at CpG sites, this modification was found, and it became reduced in two-cell stage embryos and blastocysts. These observations suggest that the process of establishment of the methylation imprint at this locus is more dynamic than previously thought. PMID- 15778221 TI - Trafficking of the plasma membrane gamma-aminobutyric acid transporter GAT1. Size and rates of an acutely recycling pool. AB - Plasma membrane neurotransmitter transporters rapidly traffic to and from the cell surface in neurons. This trafficking may be important in regulating neuronal signaling. Such regulation will be subject to the number of trafficking transporters and their trafficking rates. In the present study, we define an acutely recycling pool of endogenous gamma-aminobutyric acid transporters (GAT1) in cortical neurons that comprises approximately one-third of total cellular GAT1. Kinetic analysis of this pool estimates exocytosis and endocytosis time constants of 1.6 and 0.9 min, respectively, and thus approximately one-third of the recycling pool is plasma membrane resident in the basal state. Recent evidence shows that GAT1 substrates, second messengers, and interacting proteins regulate GAT1 trafficking. These triggers could act by altering trafficking rates or by changing the recycling pool size. In the present study we examine three GAT1 modulators. Calcium depletion decreases GAT1 surface expression by diminishing the recycling pool size. Sucrose increases GAT1 surface expression by blocking clathrin- and dynamin-dependent endocytosis, but it does not change the recycling pool size. Protein kinase C decreases surface GAT1 expression by increasing the endocytosis rate, but it does not change the exocytosis rate or the recycling pool size. Based upon estimates of GAT1 molecules in cortical boutons, the present data suggest that approximately 1000 transporters comprise the acutely recycling pool, of which 300 are on the surface in the basal state, and five transporters insert into the plasma membrane every second. This insertion could represent the fusion of one transporter-containing vesicle. PMID- 15778222 TI - Essential roles of Atg5 and FADD in autophagic cell death: dissection of autophagic cell death into vacuole formation and cell death. AB - Autophagic cell death is characterized by the accumulation of vacuoles in physiological and pathological conditions. However, its molecular event is unknown. Here, we show that Atg5, which is known to function in autophagy, contributes to autophagic cell death by interacting with Fas-associated protein with death domain (FADD). Down-regulation of Atg5 expression in HeLa cells suppresses cell death and vacuole formation induced by IFN-gamma. Inversely, ectopic expression of Atg5 using adenoviral delivery induces autophagic cell death. Deletion mapping analysis indicates that procell death activity resides in the middle and C-terminal region of Atg5. Cells harboring the accumulated vacuoles triggered by IFN-gamma or Atg5 expression become dead, and vacuole formation precedes cell death. 3-Methyladenine or expression of Atg5(K130R) mutant blocks both cell death and vacuole formation triggered by IFN-gamma, whereas benzyloxycarbonyl-VAD-fluoromethyl ketone (Z-VAD-fmk) inhibits only cell death but not vacuole formation. Atg5 interacts with FADD via death domain in vitro and in vivo, and the Atg5-mediated cell death, but not vacuole formation, is blocked in FADD-deficient cells. These results suggest that Atg5 plays a crucial role in IFN-gamma-induced autophagic cell death by interacting with FADD. PMID- 15778223 TI - Tpl2 (tumor progression locus 2) phosphorylation at Thr290 is induced by lipopolysaccharide via an Ikappa-B Kinase-beta-dependent pathway and is required for Tpl2 activation by external signals. AB - The serine-threonine protein kinase encoded by the tumor progression locus 2 (Tpl2) proto-oncogene transduces Toll-like receptor and death receptor signals in a variety of cell types. Here we show that Tpl2 undergoes phosphorylation at Thr(290) both in cells overexpressing Tpl2 and in cells stimulated with lipopolysaccharide (LPS) or tumor necrosis factor-alpha and that phosphorylation on this site parallels Tpl2 activation. Reconstitution of Tpl2(-/-) macrophages with wild type Tpl2 or Tpl2 T290D restored ERK activation by LPS, whereas reconstitution of the same cells with Tpl2 T290A did not, suggesting that phosphorylation at Thr(290) is required for the physiological activation of Tpl2 by external signals. Both the wild type Tpl2 and the kinase-inactive mutant Tpl2 K167M undergo Thr(290) phosphorylation, suggesting that Thr(290) may be a site of trans-phosphorylation rather than auto-phosphorylation. Pretreatment of 293 cells and primary macrophages with the Ikappa-B kinase-beta (IKKbeta) inhibitor PS-1145 blocked Tpl2 phosphorylation at Thr(290), suggesting that phosphorylation depends on IKKbeta, an obligatory positive regulator of Tpl2. We conclude that Tpl2 phosphorylation at Thr(290) is induced by LPS, depends on IKKbeta, and is required for the physiological activation of Tpl2 by external signals. PMID- 15778224 TI - Identification of multiple genes encoding membrane proteins with undecaprenyl pyrophosphate phosphatase (UppP) activity in Escherichia coli. AB - The bacA gene product of Escherichia coli was recently purified to near homogeneity and identified as an undecaprenyl pyrophosphate phosphatase activity (El Ghachi, M., Bouhss, A., Blanot, D., and Mengin-Lecreulx, D. (2004) J. Biol. Chem. 279, 30106-30113). The enzyme function is to synthesize the carrier lipid undecaprenyl phosphate that is essential for the biosynthesis of peptidoglycan and other cell wall components. The inactivation of the chromosomal bacA gene was not lethal but led to a significant, but not total, depletion of undecaprenyl pyrophosphate phosphatase activity in E. coli membranes, suggesting that other(s) protein(s) should exist and account for the residual activity and viability of the mutant strain. Here we report that inactivation of two additional genes, ybjG and pgpB, is required to abolish growth of the bacA mutant strain. Overexpression of either of these genes, or of a fourth identified one, yeiU, is shown to result in bacitracin resistance and increased levels of undecaprenyl pyrophosphate phosphatase activity, as previously observed for bacA. A thermosensitive conditional triple mutant delta bacA,delta ybjG,delta pgpB in which the expression of bacA is impaired at 42 degrees C was constructed. This strain was shown to accumulate soluble peptidoglycan nucleotide precursors and to lyse when grown at the restrictive temperature, due to the depletion of the pool of undecaprenyl phosphate and consequent arrest of cell wall synthesis. This work provides evidence that two different classes of proteins exhibit undecaprenyl pyrophosphate phosphatase activity in E. coli and probably other bacterial species; they are the BacA enzyme and several members from a superfamily of phosphatases that, different from BacA, share in common a characteristic phosphatase sequence motif. PMID- 15778225 TI - Investigating HIV-1 polypurine tract geometry via targeted insertion of abasic lesions in the (-)-DNA template and (+)-RNA primer. AB - A variety of biochemical and structural studies indicate that two regions of the human immunodeficiency virus type 1 (HIV-1) polypurine tract (PPT)-containing RNA/DNA hybrid deviate from standard Watson-Crick geometry. However, it is unclear whether and how these regions cooperate to ensure PPT primer selection by reverse transcriptase-associated ribonuclease H and subsequent removal from nascent (+)-DNA. To address these issues, we synthesized oligonucleotides containing abasic lesions in either the PPT (+)-RNA primer or (-)-DNA template to locally remove nucleobases, although retaining the sugar-phosphate backbone. KMnO(4) footprinting indicates such lesions locally alter duplex structure, whereas thermal melting studies show significantly reduced stability when lesions are positioned around the scissile bond. Substituting the (-)-DNA template between positions -15 and -13 altered cleavage specificity, whereas equivalent substitutions of the (+)-RNA had almost no effect. The unpaired base of the DNA template observed crystallographically (-11C) could also be removed without significant loss of cleavage specificity. With respect to the scissile -1/+1 phosphodiester bond, template nucleobases could be removed without loss of cleavage specificity, whereas equivalent lesions in the RNA primer were inhibitory. Our data suggest an interaction between the p66 thumb subdomain of HIV-1 reverse transcriptase, and the DNA template in the "unzipped" portion of the RNA/DNA hybrid could aid in positioning the ribonuclease H catalytic center at the PPT/U3 junction and also provides insights into nucleic acid geometry around the scissile bond required for hydrolysis. PMID- 15778226 TI - Phosphorylation of rat liver mitochondrial glycerol-3-phosphate acyltransferase by casein kinase 2. AB - We have previously shown rat liver mitochondrial glycerol-3-phosphate acyltransferase (mtGAT), which catalyzes the first step in de novo glycerolipid biosynthesis, is stimulated by casein kinase 2 (CK2) and that a phosphorylated protein of approximately 85 kDa is present in CK2-treated mitochondria. In this paper, we have identified the (32)P-labeled 85-kDa protein as mtGAT. We have also investigated whether the phosphorylation of mtGAT is because of CK2. Mitochondria were treated with CK2 and [gamma-(32)P]GTP as the phosphate donor. Autoradiography, Western blot, and immunoprecipitation results showed mtGAT was phosphorylated by CK2. Next, we incubated mitochondria with CK2 and either ATP or GTP, in the presence of heparin, a known inhibitor of CK2. Heparin inhibited CK2 induced stimulation of mtGAT activity; this inhibition resulted in decreased (32)P-labeling of mtGAT. Additionally, mitochondria were treated with CK2 and [gamma-(32)P]ATP in the presence of staurosporine (a serine/threonine protein kinase inhibitor), genistein (a tyrosine kinase inhibitor), and 5,6-dichloro-1 beta-D-ribofuranosylbenzimidazole (DRB, a CK2 inhibitor). Only DRB, the CK2 inhibitor, greatly reduced the amount of (32)P-incorporation into mtGAT by CK2. Finally, isolated mitochondrial outer membrane was incubated with cytosol in the presence of [gamma-(32)P]GTP; (32)P-labeled mtGAT was detected. Collectively, these data suggest that CK2 phosphorylates mtGAT. The impact of our results in the regulation of mtGAT and other anabolic processes is discussed. PMID- 15778228 TI - Human cytomegalovirus UL84 is a phosphoprotein that exhibits UTPase activity and is a putative member of the DExD/H box family of proteins. AB - Human cytomegalovirus (HCMV) UL84 is required for lytic DNA replication and is proposed to be the key factor in initiation of viral DNA synthesis. We now show that UL84 has a high degree of homology to the DExD/H (where x can be any amino acid) box family of helicases, displays UTPase activity, and is phosphorylated at serine residues. Affinity column-purified UL84-FLAG fusion protein was used in an in vitro nucleoside triphosphatase (NTPase) assay to show that UL84 has NTPase activity, preferring UTP. This UTPase activity was linear with respect to enzyme concentration and slightly enhanced by the addition of nucleic acid substrates. UL84 UTPase was the highest at low salt concentrations, a pH of 7.5, and a temperature of 45 degrees C. The enzyme preferred Mg2+ as the divalent cation but was also able to catalyze the UTPase reaction in the presence of Mn2+, Ca2+, and Zn2+ albeit at lower levels. The evidence presented here suggests that the UL84 UTPase activity may be part of an energy-generating system for helicase activity associated with the initiation of HCMV DNA replication. PMID- 15778229 TI - NHS smoking cessation services and smoking prevalence: observational study. PMID- 15778230 TI - A feasibility study of signed consent for the collection of patient identifiable information for a national paediatric clinical audit database. AB - OBJECTIVES: To investigate the feasibility of obtaining signed consent for submission of patient identifiable data to a national clinical audit database and to identify factors influencing the consent process and its success. DESIGN: Feasibility study. SETTING: Seven paediatric intensive care units in England. PARTICIPANTS: Parents/guardians of patients, or patients aged 12-16 years old, approached consecutively over three months for signed consent for submission of patient identifiable data to the national clinical audit database the Paediatric Intensive Care Audit Network (PICANet). MAIN OUTCOME MEASURES: The numbers and proportions of admissions for which signed consent was given, refused, or not obtained (form not returned or form partially completed but not signed), by age, sex, level of deprivation, ethnicity (South Asian or not), paediatric index of mortality score, length of hospital stay (days in paediatric intensive care). RESULTS: One unit did not start and one did not fully implement the protocol, so analysis excluded these two units. Consent was obtained for 182 of 422 admissions (43%) (range by unit 9% to 84%). Most (101/182; 55%) consents were taken by staff nurses. One refusal (0.2%) was received. Consent rates were significantly better for children who were more severely ill on admission and for hospital stays of six days or more, and significantly poorer for children aged 10-14 years. Long hospital stays and children aged 10-14 years remained significant in a stepwise regression model of the factors that were significant in the univariate model. CONCLUSION: Systematically obtaining individual signed consent for sharing patient identifiable information with an externally located clinical audit database is difficult. Obtaining such consent is unlikely to be successful unless additional resources are specifically allocated to training, staff time, and administrative support. PMID- 15778231 TI - Effects of acupuncture and stabilising exercises as adjunct to standard treatment in pregnant women with pelvic girdle pain: randomised single blind controlled trial. AB - OBJECTIVES: To compare the efficacy of standard treatment, standard treatment plus acupuncture, and standard treatment plus stabilising exercises for pelvic girdle pain during pregnancy. DESIGN: Randomised single blind controlled trial. Settings East Hospital, Gothenburg, and 27 maternity care centres in Sweden. PARTICIPANTS: 386 pregnant women with pelvic girdle pain. INTERVENTIONS: Treatment for six weeks with standard treatment (n = 130), standard treatment plus acupuncture (n = 125), or standard treatment plus stabilising exercises (n = 131). MAIN OUTCOME MEASURES: Primary outcome measure was pain (visual analogue scale); secondary outcome measure was assessment of severity of pelvic girdle pain by an independent examiner before and after treatment. RESULTS: After treatment the stabilising exercise group had less pain than the standard group in the morning (median difference = 9, 95% confidence interval 1.7 to 12.8; P = 0.0312) and in the evening (13, 2.7 to 17.5; P = 0.0245). The acupuncture group, in turn, had less pain in the evening than the stabilising exercise group (-14, 18.1 to -3.3; P = 0.0130). Furthermore, the acupuncture group had less pain than the standard treatment group in the morning (12, 5.9 to 17.3; P < 0.001) and in the evening (27, 13.3 to 29.5; P < 0.001). Attenuation of pelvic girdle pain as assessed by the independent examiner was greatest in the acupuncture group. CONCLUSION: Acupuncture and stabilising exercises constitute efficient complements to standard treatment for the management of pelvic girdle pain during pregnancy. Acupuncture was superior to stabilising exercises in this study. PMID- 15778232 TI - Opportunity cost of antidepressant prescribing in England: analysis of routine data. PMID- 15778235 TI - The workload of GPs: patients with psychological and somatic problems compared. AB - BACKGROUND: GPs state that patients with mental problems make heavy demands on their available time. To what extent these perceived problems correspond with reality needs more investigation. OBJECTIVES: To investigate the effect of patients with psychological or social diagnoses on GP's workload, expressed in time investments. METHODS: Data were derived of a cross-sectional National Survey in General Practice, conducted in The Netherlands in 2000-2002. For a year, all patient contacts with a representative sample of 104 general practices were registered. Patients diagnosed with one or more diagnoses in ICPC (International Classification of Primary Care) chapter 'Psychological' or 'Social' (n = 37,189) were compared to patients with only somatic diagnoses (n = 189,731). A subdivision was made in diagnoses depression, anxiety, sleeping disorders, stress problems, problems related to work or partner and 'other psychological or social problems'. Workload measures are the consultation frequency, number of diagnoses and episodes of illness of the patients involved. RESULTS: Patients in all categories of psychological or social problems had almost twice as many contacts with their general practice as patients with only somatic problems. They received more diagnoses and more episodes of illness were shown. Patients with psychological or social diagnoses also contacted their general practice about their somatic problems more frequently, compared to patients with only somatic problems. CONCLUSION: Patients with psychological or social problems make heavy demands on the GP's workload, for the greater part due to the increase in somatic problems presented. PMID- 15778236 TI - The challenges of cross-national research in primary health care across Europe. PMID- 15778237 TI - Forty-eight hour access to primary care: practice factors predicting patients' perceptions. AB - BACKGROUND: The government has proposed a 48-hour target for GP availability. Although many practices are moving towards delivering that goal, recent national patient surveys have reported a deterioration in patients' reports of doctor availability. What practice factors contribute to patients' perceptions of doctor availability? METHOD: A cross sectional patient survey (11,000 patients from 54 inner London practices, 7247 (66%) respondents) using the General Practice Assessment Survey. We asked patients how soon they could be seen in their practice following non-urgent consultation requests and related their aggregated responses to the characteristics of their practice. RESULTS: Three factors relating to practice administration and appointments systems operation independently predicted patients' reports of doctor availability. These were the proportion of patients asked to attend the surgery and wait to be seen, the proportion of patients seen using an emergency surgery arrangement, and the extent of practice computerization. CONCLUSION: Some practices may have difficulty in meeting the target for GP availability. Meeting the target will involve careful review of practice administrative procedures. PMID- 15778238 TI - Magnetic resonance imaging for accelerated assessment of drug effect and prediction of subsequent radiographic progression in rheumatoid arthritis: a study of patients receiving combined anakinra and methotrexate treatment. AB - OBJECTIVES: By MRI to assess the efficacy of addition of anakinra for controlling synovitis and stopping erosive progression in patients with clinically active RA despite receiving methotrexate, and to determine the predictive value of MRI for subsequent radiographic erosive progression. METHODS: 100 mg anakinra subcutaneously/day was added to the treatment of 17 patients with clinically active RA despite methotrexate. MRI of the non-dominant wrist and 2nd-5th MCP joints (OMERACT evaluation) was performed at weeks 0, 12, and 36, and radiography of both hands and wrists (modified Sharp evaluation) at weeks 0 and 36. RESULTS: MRI synovitis scores were not significantly changed. Radiography of both hands and wrists after 36 weeks showed erosive progression in 11 patients, and MRI after 12 weeks in 10 patients. Nine of 10 patients with MRI progression at 12 weeks had radiographic progression at 36 weeks. Baseline MRI synovitis and erosion scores, but no clinical/biochemical parameters, correlated significantly with subsequent erosive progression. CONCLUSION: Addition of anakinra did not significantly reduce MRI signs of synovitis, and most patients had progressive joint destruction. Baseline MRI findings predicted subsequent radiographic erosive progression. Unilateral wrist and MCP joint MRI after 12 weeks had a similar sensitivity for detection of erosive progression as bilateral hand and wrist radiography after 36 weeks. PMID- 15778239 TI - Transdifferentiation of polymorphonuclear neutrophils to dendritic-like cells at the site of inflammation in rheumatoid arthritis: evidence for activation by T cells. AB - OBJECTIVES: To investigate infiltrated cells in the synovial fluid (SF) of inflamed joints of patients with rheumatoid arthritis (RA), with special reference to polymorphonuclear neutrophils (PMN) and their interaction with T cells. METHODS: Expression on PMN of activation associated receptors CD14, CD64, CD83, and major histocompatibility complex (MHC) class II was examined in the SF of 15 patients with RA, as were the infiltrated T cells. SF cytokines were determined by enzyme linked immunosorbent assay (ELISA). To mimic the in vivo situation, co-culture experiments were carried out using PMN and T cells of healthy donors. RESULTS: The SF contained activated T lymphocytes and abundant PMN. SF PMN expression of CD14 and CD64 was enhanced compared with peripheral blood. Of special interest was the observation that only the SF PMN expressed MHC class II antigens and CD83. Exposure to SF, which contained considerable amounts of cytokines (for example, interferon gamma (IFNgamma), tumour necrosis factor alpha, and interleukin 2), induced a similar receptor pattern on blood derived PMN of healthy donors. Furthermore, PMN acquired MHC class II and CD83 within 24 to 48 hours, when co-cultured with autologous T cells or T cell lines. This effect was also achieved by T cell supernatants, was dependent on protein synthesis, and could be inhibited by antibodies against IFNgamma. CONCLUSIONS: SF PMN from patients with RA undergo major alterations, including transdifferentiation to cells with dendritic-like characteristics, probably induced by T cell derived cytokines. Because MHC class II positive PMN are known to activate T cells, the mutual activation of PMN and T cells might contribute to the perpetuation of the local inflammatory process, and eventually to the destructive process in RA. PMID- 15778240 TI - Radiographic progression in patients with ankylosing spondylitis after 2 years of treatment with the tumour necrosis factor alpha antibody infliximab. AB - BACKGROUND: Anti-tumour necrosis factor (TNF) treatment is clinically efficacious in patients with active ankylosing spondylitis (AS) and leads to improvement of spinal inflammation, as assessed by magnetic resonance imaging. It is unclear whether anti-TNF treatment affects chronic spinal changes in AS. OBJECTIVES: To analyse the effect of infliximab on the radiographic course of AS over 2 years. METHODS: Complete sets of lateral radiographs of the cervical spine and lumbar spine were available from 82 patients from two sources: 41 patients (group 1) had been treated with infliximab (5 mg/kg/6 weeks) as part of a recent randomised controlled trial and 41 patients (group 2) were part of the early German AS cohort (GESPIC), without controlled interventions. Radiographs were obtained at baseline and after 2 years and scored by the modified Stokes AS Spinal Score (mSASSS). RESULTS: Patients in the infliximab group were older, had a longer disease duration, and more radiographic damage at baseline. The mean (SD) mSASSS change was 0.4 (2.7) and 0.7 (2.8) for groups 1 and 2, respectively (p = NS). Radiographic damage at baseline was a predictor for more radiographic progression. Patients with baseline damage who were treated with infliximab showed a trend for less radiographic progression. No correlations between clinical parameters and radiographic progression were found. CONCLUSIONS: Patients with AS treated with infliximab had less radiographic progression after 2 years. Patients with prevalent radiographic damage are prone to develop more damage over time. Infliximab may decelerate radiographic progression in such patients. Larger studies are needed to prove that anti-TNF treatment inhibits structural damage. PMID- 15778241 TI - Impact of pregnancy on health related quality of life evaluated prospectively in pregnant women with rheumatic diseases by the SF-36 health survey. AB - OBJECTIVE: To gain insight into patient experience of the disease course and health related quality of life during and after pregnancy in women with rheumatoid arthritis and ankylosing spondylitis. METHODS: 10 patients with rheumatoid arthritis, 10 patients with ankylosing spondylitis, and 29 age matched healthy pregnant controls were evaluated by the medical outcomes study short form 36 (SF-36) health survey once at each trimester and at 6, 12, and 24 weeks postpartum. A group of non-pregnant age matched female patients (40 rheumatoid arthritis, 16 ankylosing spondylitis) was studied for comparison. RESULTS: Impaired physical dimensions as well as increased bodily pain was observed in healthy women in late pregnancy. Patients with rheumatoid arthritis showed improved physical functioning scores in the second trimester and reduced pain in the third trimester. Among pregnant patients, those with ankylosing spondylitis suffered the greatest impairment of health related quality of life during pregnancy. In all patient groups the physical impairment in the third trimester was less pronounced than in healthy controls. Mental health scores remained stable even with persisting active disease during pregnancy, or with a postpartum flare. CONCLUSIONS: Pregnancy reduced physical functioning in healthy women and patients, but had no impact on mental and emotional health, even at times of disease aggravation. The pregnancy experience documented in our patients may be helpful when counselling patients contemplating pregnancy. PMID- 15778242 TI - Close association between valvar heart disease and central nervous system manifestations in the antiphospholipid syndrome. AB - BACKGROUND: Heart valves lesions and central nervous system involvement are among the most common manifestations of the antiphospholipid syndrome (APS). OBJECTIVE: To evaluate possible interrelations between these manifestations in a large group of APS patients. METHODS: 284 APS patients were evaluated retrospectively, 159 of whom had primary APS. Cardiac-CNS associations were determined for the entire study population, and for subgroups of patients with primary APS or APS associated with systemic lupus erythematosus (SLE). RESULTS: Significant associations where found between cardiac vegetations and epilepsy (p < 0.02), and between cardiac valve thickening or dysfunction and migraine (p = 0.002). Borderline association was found between valvar vegetations and migraine (p = 0.09). A significant association was also found between all valvar lesions and stroke or transient ischaemic attacks. Subanalyses showed that patients with primary APS had significant associations between cardiac valve pathology and all CNS manifestations, while patients with APS associated with SLE had no such associations. CONCLUSIONS: The study suggests potential differences in biological behaviour between primary APS and APS associated with SLE. The presence of cardiac valve pathology may be a risk factor for several types of CNS involvement in PAPS. PMID- 15778243 TI - Magnetic resonance imaging of the spine and the sacroiliac joints in ankylosing spondylitis and undifferentiated spondyloarthritis during treatment with etanercept. AB - OBJECTIVE: To assess the changes in inflammatory lesions of the spine and the sacroiliac (SI) joints as detected by magnetic resonance imaging (MRI) in patients with ankylosing spondylitis (AS) and undifferentiated spondyloarthritis (uSpA) with predominant axial symptoms during treatment with etanercept. METHODS: MRI of the spine and/or the SI joints of patients with active AS or axial uSpA was performed at baseline (TP0, n = 25), after 6 weeks (TP1, n = 20), and after 24 weeks of continuous treatment with etanercept (TP2, n = 12). T1 weighted spin echo pre -(T1), post-gadolinium (T1/Gd-DTPA) and short tau inversion recovery (STIR) MRI sequences were used to assess chronic and active spinal lesions using the scoring system ASspiMRI. Active and chronic SI lesions were assessed using a simple scoring system. RESULTS: By use of the definite STIR sequence, significant regression of spinal inflammation was already seen already after 6 weeks in the patients treated with etanercept (mean (SD) 11.2 (13.8) at TP0 v 6.8 (7.9) at TP1; p = 0.023) but not in patients treated with placebo. Continuous treatment with etanercept for 24 weeks reduced active spinal changes by 69% (p = 0.012). T1/Gd-DTPA sequences gave similar results. There was only a trend for a decrease of active inflammatory lesions of the SI joints. CONCLUSIONS: Etanercept treatment in patients with active AS and uSpA leads to regression of active inflammatory lesions of the spine as depicted by MRI. The potential role of etanercept on deceleration of chronic spinal changes needs further study. PMID- 15778244 TI - Effect of doxycycline on sulfur mustard-induced respiratory lesions in guinea pigs. AB - Respiratory tract lesions induced by the chemical warfare agent sulfur mustard (SM) are characterized by epithelial damages associated with inflammatory cell infiltration. Here we evaluated the imbalance between gelatinase and tissue inhibitors of metalloproteinases (TIMPs), and we tested pretreatment with the protease inhibitor doxycycline. Guinea pigs were intoxicated intratracheally with SM and evaluated 24 h after exposure. Matrix metalloproteinase (MMP) gelatinase activity of bronchial lavage (BL) fluid from SM-exposed guinea pigs was high compared with controls, as shown by both zymography and biotinylated substrate degradation, whereas TIMP-1 and -2 levels by immunoblotting were similar. Extensive areas of lysis were evidenced by in situ zymography, indicating imbalance between gelatinases and inhibitors towards net proteolytic activity. Doxycycline pretreatment resulted in 1) decreased gelatinase activity (zymography, free gelatinase activity assay, and in situ zymography); 2) decreased inflammation (BL fluid cellularity and protein level); and 3) dramatic decrease in histological epithelial lesions. Our results suggest inadequate levels of TIMP to counteract increased gelatinase activity and further support a role for MMP gelatinases in SM-induced respiratory lesions. They also suggest that doxycycline may hold promise as a therapeutic tool. PMID- 15778245 TI - MAP kinases in lung endothelial permeability induced by microtubule disassembly. AB - Lung endothelial barrier function is regulated by multiple signaling pathways, including mitogen-activated protein kinases (MAPK) extracellular signal-regulated kinases (ERK) 1/2 and p38. We have recently shown involvement of microtubule (MT) disassembly in endothelial cell (EC) barrier failure. In this study, we examined potential involvement of ERK1/2 and p38 MAPK in lung EC barrier dysfunction associated with MT disassembly. MT inhibitors nocodazole (0.2 microM) and vinblastine (0.1 microM) induced sustained activation of Ras-Raf-MEK1/2-ERK1/2 and MKK3/6-p38-MAPKAPK2 MAPK cascades in human and bovine pulmonary EC, as detected by phosphospecific antibodies and in MAPK activation assays. These effects were linked to increased permeability assessed by measurements of transendothelial electrical resistance and cytoskeletal remodeling analyzed by morphometric analysis of EC monolayers. MT stabilization by taxol (5 microM, 1 h) attenuated nocodazole-induced ERK1/2 and p38 MAPK activation and phosphorylation of p38 MAPK substrate 27-kDa heat shock protein and regulatory myosin light chains, the proteins involved in actin polymerization and actomyosin contraction. Importantly, only pharmacological inhibition of p38 MAPK by SB-203580 (20 microM, 1 h) attenuated nocodazole-induced MT depolymerization, actin remodeling, and EC barrier dysfunction, whereas the MEK/ERK1/2 inhibitor U0126 (5 microM, 1 h) exhibited no effect. These data suggest a direct link between p38 MAPK activation, remodeling of MT network, and EC barrier regulation. PMID- 15778246 TI - Effect of CO2 on LPS-induced cytokine responses in rat alveolar macrophages. AB - Alveolar macrophages (AM) may be exposed to a range of CO(2) and pH levels depending on their location in the alveoli and the health of the lung. Cytokines produced by AM contribute to inflammation in acute lung injury (ALI). Current ventilatory practices for the management of ALI favor low tidal volumes, which can give rise to increases in CO(2) and changes in pH of the alveolar microenvironment. Here we examined the effect of CO(2) on cytokine release from LPS-stimulated rat AM. AM were incubated for 1-4 h under different atmospheric gas mixtures ranging from 2.5-20% CO(2). To distinguish between effects of pH and CO(2), the culture media were also buffered to pH 7.2 with NaHCO(3). Cell metabolic activity, but not cell viability, decreased and increased significantly after 4 h at 20 and 2.5% CO(2), respectively. Increasing CO(2) decreased TNF alpha secretion but had no effect on lysate TNF-alpha. Buffering the media abated the effects of CO(2) on TNF-alpha secretion. CO(2) increased cytokine-induced neutrophil chemoattractant factor-1 secretion only when the pH was buffered to 7.2. Effects of CO(2) on cytokine responses were reversible. In conclusion, the effects of CO(2) on cytokine lysate levels and/or secretion in AM are cytokine specific and, depending on both the cytokine and the immediate microenvironment, may be beneficial or detrimental to ALI. PMID- 15778247 TI - Acid and particulate-induced aspiration lung injury in mice: importance of MCP-1. AB - A model of aspiration lung injury was developed in WT C57BL/6 mice to exploit genetically modified animals on this background, i.e., MCP-1(-/-) mice. Mice were given intratracheal hydrochloric acid (ACID, pH 1.25), small nonacidified gastric particles (SNAP), or combined acid plus small gastric particles (CASP). As reported previously in rats, lung injury in WT mice was most severe for "two-hit" aspiration from CASP (40 mg/ml particulates) based on the levels of albumin, leukocytes, TNF-alpha, IL-1beta, IL-6, MCP-1, KC, and MIP-2 in bronchoalveolar lavage (BAL) at 5, 24, and 48 h. MCP-1(-/-) mice given 40 mg/ml CASP had significantly decreased survival compared with WT mice (32% vs. 80% survival at 24 h and 0% vs. 72% survival at 48 h). MCP-1(-/-) mice also had decreased survival compared with WT mice for CASP aspirates containing reduced particulate doses of 10-20 mg/ml. MCP-1(-/-) mice given 5 mg/ml CASP had survival similar to WT mice given 40 mg/ml CASP. MCP-1(-/-) mice also had differing responses from WT mice for several inflammatory mediators in BAL (KC or IL-6 depending on the particle dose of CASP and time of injury). Histopathology of WT mice with CASP (40 mg particles/ml) showed microscopic areas of compartmentalization with prominent granuloma formation by 24 h, whereas lung tissue from MCP-1(-/-) mice had severe diffuse pneumonia without granulomas. These results indicate that MCP 1 is important for survival in murine aspiration pneumonitis and appears to act partly to protect uninjured lung regions by promoting isolation and compartmentalization of tissue with active inflammation. PMID- 15778248 TI - Enhanced susceptibility of staggerer (RORalphasg/sg) mice to lipopolysaccharide induced lung inflammation. AB - The retinoid-related orphan receptor alpha (RORalpha), a member of the ROR subfamily of nuclear receptors, has been implicated in the control of a number of physiological processes, including the regulation of several immune functions. To study the potential role of RORalpha in the regulation of innate immune responses in vivo, we analyzed the induction of airway inflammation in response to lipopolysaccharide (LPS) challenge in wild-type and staggerer (RORalpha(sg/sg)) mice, a natural mutant strain lacking RORalpha expression. Examination of hematoxylin and eosin-stained lung sections showed that RORalpha(sg/sg) mice displayed a higher degree of LPS-induced inflammation than wild-type mice. Bronchoalveolar lavage (BAL) was performed at 3, 16, and 24 h after LPS exposure to monitor the increase in inflammatory cells and the level of several cytokines/chemokines. The increased susceptibility of RORalpha(sg/sg) mice to LPS induced airway inflammation correlated with a higher number of total cells and neutrophils in BAL fluids from LPS-treated RORalpha(sg/sg) mice compared with those from LPS-treated wild-type mice. In addition, IL-1beta, IL-6, and macrophage inflammatory protein-2 were appreciably more elevated in BAL fluids from LPS-treated RORalpha(sg/sg) mice compared with those from LPS-treated wild type mice. The enhanced susceptibility of RORalpha(sg/sg) mice appeared not to be due to a repression of IkappaBalpha expression. Our observations indicate that RORalpha(sg/sg) mice are more susceptible to LPS-induced airway inflammation and are in agreement with the hypothesis that RORalpha functions as a negative regulator of LPS-induced inflammatory responses. PMID- 15778249 TI - Mycoplasma pneumoniae infection increases airway collagen deposition in a murine model of allergic airway inflammation. AB - Mycoplasma pneumoniae (Mp) has been linked to chronic asthma. Airway remodeling (e.g., airway collagen deposition or fibrosis) is one of the pathological features of chronic asthma. However, the effects of respiratory Mp infection on airway fibrosis in asthma remain unclear. In the present study, we hypothesized that respiratory Mp infection may increase the airway collagen deposition in a murine model of allergic airway inflammation in part through upregulation of transforming growth factor (TGF)-beta1. Double (2 wk apart) inoculations of Mp or saline (control) were given to mice with or without previous allergen (ovalbumin) challenges. On days 14 and 42 after the last Mp or saline, lung tissue and bronchoalveolar lavage (BAL) fluid were collected for analyses of collagen and TGF-beta1 at protein and mRNA levels. In allergen-naive mice, Mp did not alter airway wall collagen. In allergen-challenged mice, Mp infections did not change airway wall collagen deposition on day 14 but increased the airway collagen on day 42; this increase was accompanied by increased TGF-beta1 protein in the airway wall and reduced TGF-beta1 protein release from the lung tissue into BAL fluid. Our results suggest that Mp infections could modulate airway collagen deposition in a murine model of allergic airway inflammation with TGF-beta1 involved in the collagen deposition process. PMID- 15778250 TI - Farming and asthma. PMID- 15778251 TI - The role of occupation in the development of chronic obstructive pulmonary disease (COPD). PMID- 15778252 TI - Occupation and COPD. PMID- 15778253 TI - Exposure to traffic exhausts and oxidative DNA damage. AB - AIMS: To assess the relations between exposure to traffic exhausts and indicators of oxidative DNA damage among highway toll station workers. METHODS: Cross sectional study of 47 female highway toll station workers exposed to traffic exhausts and 27 female office workers as a reference group. Exposure assessment was based on average and cumulative traffic density and a biomarker of exposure, urinary 1-hydroxypyrene-glucuronide (1-OHPG). Urinary 8-hydroxydeoxyguanosine (8 OHdG) was used as a biomarker of oxidative DNA damage. Plasma nitric oxide (NO) was measured as an indicator of oxidative stress related to traffic exhaust exposure. RESULTS: The mean concentration of urinary 8-OHdG was substantially higher among the exposed non-smokers (13.6 microg/g creatinine) compared with the reference non-smokers (7.3 microg/g creatinine; difference 6.3, 95% CI 3.0 to 9.6). The mean concentration of NO among the exposed (48.0 micromol/l) was also higher compared with the reference non-smokers (37.6 micromol/l; difference 10.4, 95% CI -0.4 to 21.2). In linear regression adjusting for confounding, a change in log(8-OHdG) was statistically significantly related to a unit change in log(1 OHPG) (beta = 0.372, 95% CI 0.081 to 0.663). CONCLUSIONS: Results indicate that exposure to traffic exhausts increases oxidative DNA damage. Urinary 8-OHdG is a promising biomarker of traffic exhaust induced oxidative stress. PMID- 15778254 TI - Do changes in effort-reward imbalance at work contribute to an explanation of the social gradient in angina? AB - AIMS: To determine whether an increase in effort-reward imbalance over time increases the risk of angina, and whether such increases are associated with lower occupational position. METHODS: Effort-reward imbalance (ERI) at work was measured in the Whitehall II occupational cohort of London based civil servants at baseline (1985-88) and in 1997. Coronary heart disease was measured in a self reported health questionnaire by combining the Rose Angina Questionnaire with doctor diagnosed angina in 2001. RESULTS: Among men, increase in ERI over time was associated with an increased risk of incident angina. Moreover, as increases in ERI were more common among lower grade civil servants, change in imbalance, to some extent, contributed to explaining the social gradient in angina. Among women, increases in imbalance were not associated with risk of angina, and therefore did not contribute to the explanation of the social gradient. CONCLUSIONS: Reductions in effort-reward imbalance at work may reduce the risk of coronary heart disease among men. PMID- 15778255 TI - Cancer risks in a historical UK cohort of benzene exposed workers. AB - AIMS: To examine mortality from different causes and cancer incidence among a cohort of benzene workers in England and Wales. METHODS: A cohort of 5514 workers who had been occupationally exposed to benzene in 1966/67 or earlier was assembled by the former Factory Inspectorate and the Medical Research Council from details provided by 233 employers in England and Wales. The cohort was followed up for mortality (1968-2002) and cancer registrations (1971-2001). National mortality rates and cancer registration (incidence) rates were used to calculate standardised mortality ratios and standardised registration ratios. RESULTS: Mortality was close to expectation for all causes and significantly increased for cancer of the lip, cancer of the lung and bronchus, secondary and unspecified cancers, acute non-lymphocytic leukaemia (ANLL), and all neoplasms. Significant deficits were shown for three non-malignant categories (mental disorders, diseases of the digestive system, accidents). SMRs for other leukaemia, lymphomas, and multiple myeloma were close to or below expectation. There was some evidence of under-ascertainment of cancer registrations, although significantly increased SRRs were shown for lung cancer and cancer of the pleura (mesothelioma). CONCLUSIONS: Many study subjects would have been exposed to carcinogens other than benzene (for example, asbestos, rubber industry fumes, foundry fumes, polycyclic aromatic hydrocarbons), and the excesses of lung cancer and mesothelioma are likely to reflect exposures to these other carcinogens. The carcinogenic effects of benzene exposure on the lymphohaematopoietic system were limited to ANLL. PMID- 15778256 TI - A follow up study of vibration induced white finger in compensation claimants. AB - AIMS: To follow up vibration induced white finger (VWF) in a selected group of 73 vibration exposed workers who claimed unsuccessfully for VWF compensation at a first examination. METHODS: The VWF claimants were sent to our unit by the National Insurance Institute. The basic compensatory criteria included a positive history of VWF and abnormal cold response of the digital arteries. Following the first unsuccessful examination, over a mean time period of 4.1 (range 1-11) years the National Insurance Institute requested a second examination for all 73 claimants and a third examination for 29. During the follow up period, all subjects continued to work with vibratory tools. RESULTS: There were 14 new cases who reported white finger during the follow up period. In the new VWF cases, finger blanching attacks became visible after about 3.5 years since the first examination. All incident cases of anamnestic VWF showed an abnormal cold response in the digital arteries and obtained compensation according to the basic compensatory criteria. In the entire sample of VWF claimants, there was a discrepancy between positive history of VWF symptoms at medical interview (55%) and abnormal cold provocation outcomes (19%). Digital arterial hyperresponsiveness to cold was associated with both VWF symptoms and the duration of vibration exposure since the first examination. Over the follow up period, a significant increase in the vasoconstrictor response to cold was observed in the vibration exposed workers with no symptoms of finger whiteness. Abnormal cold response was not associated with either age or smoking habit. CONCLUSIONS: Cold test measuring finger systolic blood pressure may be considered a useful laboratory method to confirm objectively VWF symptoms and to disclose abnormal cold induced vasoconstrictor response in vibration exposed workers with a negative history of VWF. Medical interview outcomes should be interpreted with caution in medicolegal situations involving VWF claimants. PMID- 15778257 TI - Relationship between chemical structure and the occupational asthma hazard of low molecular weight organic compounds. AB - AIMS: To investigate quantitatively, relationships between chemical structure and reported occupational asthma hazard for low molecular weight (LMW) organic compounds; to develop and validate a model linking asthma hazard with chemical substructure; and to generate mechanistic hypotheses that might explain the relationships. METHODS: A learning dataset used 78 LMW chemical asthmagens reported in the literature before 1995, and 301 control compounds with recognised occupational exposures and hazards other than respiratory sensitisation. The chemical structures of the asthmagens and control compounds were characterised by the presence of chemical substructure fragments. Odds ratios were calculated for these fragments to determine which were associated with a likelihood of being reported as an occupational asthmagen. Logistic regression modelling was used to identify the independent contribution of these substructures. A post-1995 set of 21 asthmagens and 77 controls were selected to externally validate the model. RESULTS: Nitrogen or oxygen containing functional groups such as isocyanate, amine, acid anhydride, and carbonyl were associated with an occupational asthma hazard, particularly when the functional group was present twice or more in the same molecule. A logistic regression model using only statistically significant independent variables for occupational asthma hazard correctly assigned 90% of the model development set. The external validation showed a sensitivity of 86% and specificity of 99%. CONCLUSIONS: Although a wide variety of chemical structures are associated with occupational asthma, bifunctional reactivity is strongly associated with occupational asthma hazard across a range of chemical substructures. This suggests that chemical cross-linking is an important molecular mechanism leading to the development of occupational asthma. The logistic regression model is freely available on the internet and may offer a useful but inexpensive adjunct to the prediction of occupational asthma hazard. PMID- 15778258 TI - Impact of occupational stress and other psychosocial factors on musculoskeletal pain among Chinese offshore oil installation workers. AB - AIMS: To explore the relation between psychosocial factors and musculoskeletal pain in Chinese offshore oil installation workers. METHODS: Half of all offshore workers (being a representative sample) in a Chinese oil company were invited to complete a self-administered questionnaire providing information on sociodemographic characteristics, occupational stressors, type A behaviour, social support, coping style, health related behaviour, past injuries, and musculoskeletal pain. Factor analysis was used to identify the sources of occupational stress and the domains of type A behaviour and coping style. Logistic regression analyses were used to study the relations between psychosocial factors and musculoskeletal pain in each body region. RESULTS: The prevalence of musculoskeletal pain over the previous 12 months varied between 7.5% for elbow pain and 32% for low back pain; 56% workers had at least one complaint. Significant associations were found between various psychosocial factors and musculoskeletal pain in different body regions after adjusting for potential confounding factors. Occupational stressors, in particular stress from safety, physical environment, and ergonomics, were important predictors of musculoskeletal pain, as was coping by eating behaviour. CONCLUSIONS: These observations supported the widely accepted biopsychosocial model of musculoskeletal disorders and suggested that in future studies of work related musculoskeletal disorders, psychosocial factors must be given due consideration. PMID- 15778259 TI - Computer based safety training: an investigation of methods. AB - BACKGROUND: Computer based methods are increasingly being used for training workers, although our understanding of how to structure this training has not kept pace with the changing abilities of computers. Information on a computer can be presented in many different ways and the style of presentation can greatly affect learning outcomes and the effectiveness of the learning intervention. Many questions about how adults learn from different types of presentations and which methods best support learning remain unanswered. AIMS: To determine if computer based methods, which have been shown to be effective on younger students, can also be an effective method for older workers in occupational health and safety training. METHODS: Three versions of a computer based respirator training module were developed and presented to manufacturing workers: one consisting of text only; one with text, pictures, and animation; and one with narration, pictures, and animation. After instruction, participants were given two tests: a multiple choice test measuring low level, rote learning; and a transfer test measuring higher level learning. RESULTS: Participants receiving the concurrent narration with pictures and animation scored significantly higher on the transfer test than did workers receiving the other two types of instruction. There were no significant differences between groups on the multiple choice test. CONCLUSIONS: Narration with pictures and text may be a more effective method for training workers about respirator safety than other popular methods of computer based training. Further study is needed to determine the conditions for the effective use of this technology. PMID- 15778261 TI - Increased incidence of cutaneous malignant melanoma among longshoremen in Genoa, Italy: the role of sunlight and occupational exposure. PMID- 15778260 TI - Factors influencing the difference between maternal and cord blood lead. AB - AIMS: To determine the factors that affect why some infants receive higher exposures relative to the mother's body burden than do others. METHODS: A total of 159 mother-infant pairs from a cohort of women receiving prenatal care at Magee-Womens Hospital in Pittsburgh, PA from 1992 to 1995 provided blood samples at delivery for lead determination. The difference between cord and maternal blood lead concentration (PbB) and a dichotomous variable indicator of higher cord than maternal PbB, were examined as indicators of relative transfer. Women were interviewed twice during the pregnancy about lifestyle, medical history, calcium nutrition, and physical activity. RESULTS: Higher blood pressure was associated with relatively greater cord compared with maternal PbB, as was maternal alcohol use. Sickle cell trait and higher haemoglobin were associated with a lower cord relative to maternal blood lead PbB. No association was seen with smoking, physical exertion, or calcium consumption. CONCLUSION: While reduction in maternal exposure will reduce fetal exposure, it may also be possible to mitigate infant lead exposure by reducing transfer from the pregnant woman. Interventions aimed at reducing blood pressure and alcohol consumption during pregnancy may be useful in this regard. PMID- 15778262 TI - Changing trends in US mesothelioma incidence. AB - The correspondence by Weillet al (below) refers to a letter by Greenberg, which was published in February's edition of the journal. We regret the late appearance of this printed response, which arises from an administrative error. An electronic version of this text was posted on the website on 1 February 2005. PMID- 15778263 TI - Design of exposure questionnaires for epidemiological studies. PMID- 15778264 TI - Dynamic association of nitric oxide downstream signaling molecules with endothelial caveolin-1 in rat aorta. AB - Classically, nitric oxide (NO) formed by endothelial NO synthase (eNOS) freely diffuses from its generation site to smooth muscle cells where it activates soluble guanylyl cyclase (sGC), producing cGMP. Subsequently, cGMP activates both cGMP- and cAMP-dependent protein kinases [cGMP-dependent protein kinase (PKG) and cAMP-dependent protein kinase (PKA), respectively], leading to smooth muscle relaxation. In endothelial cells, eNOS has been localized to caveolae, small invaginations of the plasma membrane rich in cholesterol. Membrane cholesterol depletion impairs acetylcholine (ACh)-induced relaxation due to alteration in caveolar structure. Given the nature of NO to be more soluble in a hydrophobic environment than in water, and assuming that colocalization of components in a signal transduction cascade seems to be a critical determinant of signaling efficiency by eNOS activation, we hypothesize that sGC, PKA, and PKG activation may occur at the plasma membrane caveolae. In endothelium-intact rat aortic rings, the relaxation induced by ACh, by the sGC activator 3-(5'-hydroxymethyl 2'furyl)-1-benzyl indazole (YC-1), and by 8-bromo-cGMP was impaired in the presence of methyl-beta-cyclodextrin, a drug that disassembles caveolae by sequestering cholesterol from the membrane. sGC, PKG, and PKA were colocalized with caveolin-1 in aortic endothelium, and this colocalization was abolished by methyl-beta-cyclodextrin. Methyl-beta-cyclodextrin efficiently disassembled caveolae in endothelium. In summary, our results provide evidence of compartmentalization of sGC, PKG, and PKA in endothelial caveolae contributing to NO signaling cascade, giving new insights by which the endothelium mediates vascular smooth muscle relaxation. PMID- 15778265 TI - A novel inhibitor of N-ethylmaleimide-sensitive factor decreases leukocyte trafficking and peritonitis. AB - Endothelial exocytosis is an early stage in the process of leukocyte trafficking. N-ethylmaleimide-sensitive factor (NSF) plays a critical role in regulating exocytosis. We hypothesized that inhibitors of NSF decrease endothelial exocytosis and vascular inflammation. We designed a novel fusion polypeptide consisting of a human immunodeficiency virus transactivator of transcription (TAT) protein transduction domain joined to a NSF homohexamerization domain. We show that this TAT-NSF polypeptide inhibits the ATPase activity and the disassembly activity of NSF. Furthermore, the TAT-NSF polypeptide decreases endothelial cell exocytosis and reduces leukocyte adherence to endothelial cells in culture. Finally, the TAT-NSF polypeptide inhibits leukocyte rolling on murine venules in vivo and inhibits leukocyte trafficking into the peritoneal cavity in a murine model of experimental peritonitis. These data suggest that NSF is a critical regulator of leukocyte trafficking in vivo. Novel compounds that inhibit the exocytic machinery in endothelial cells may be useful anti-inflammatory drugs. PMID- 15778266 TI - The role of dopamine in a model of trigeminovascular nociception. AB - Migraine is a common, disabling problem with three phases: premonitory, main headache attack, and postdrome. The headache phase is thought to involve activation of trigeminal neurons, whereas the premonitory and postdrome phases may involve dopaminergic mechanisms. In animal studies, dopamine has been found to cause vasodilation of cranial arteries at very low doses. Using intravital microscopy, we examined the effect of dopamine receptor agonists on dural blood vessel caliber and the effect of dopamine and specific dopamine receptor antagonists on trigeminovascular neurogenic dural vasodilation. Dopamine hydrochloride caused a significant vasoconstriction (P < 0.05) and increase in arterial blood pressure (P < 0.05) that was reversed by a alpha2-adrenoceptor antagonist, yohimbine, rather than specific dopamine receptor antagonists. The D1 receptor agonist caused a vasoconstriction (P < 0.05) and a blood pressure increase (P < 0.05), which was reversed by yohimbine and therefore alpha2 adrenoceptor-mediated. None of the specific dopamine receptor antagonists were able to attenuate neurogenic dural vasodilation. Dopamine hydrochloride infusion (P < 0.05) and a D1 receptor agonist were able to attenuate the vasodilation (P < 0.05), with maximal dilation returning after cessation of the dopamine agonist infusion. This response may be due to the vasoconstrictor effects of the alpha2 adrenoceptor and an action at the D1 receptor. In the intravital model of trigeminal activation, it seems that dopamine receptors do not play a major role and may not present an acute treatment option. Our data do not exclude a role for dopamine receptor modulators in short- or long-term prevention. PMID- 15778267 TI - Effects of isoflurane and xenon on Ba2+-currents mediated by N-type calcium channels. AB - BACKGROUND: Isoflurane and xenon are inhalation general anaesthetics with differing clinical profiles and contrasting synaptic actions. Both agents have been shown to depress excitatory synaptic responses. Whether this is via pre synaptic or post-synaptic mechanisms has not been determined clearly. N-type calcium channels are a putative pre-synaptic target for these agents. We tested whether N-type calcium channels were sensitive to isoflurane and xenon and whether there was any stereoselectivity in the effect of isoflurane. METHODS: We used patch-clamp electrophysiology on isolated HEK293 cells stably expressing N type calcium channels to investigate the effects of isoflurane and xenon on barium currents mediated by N-type calcium channels. RESULTS: Racemic isoflurane caused a concentration-dependent reduction (11-35%) in the peak current through the N-type channels in the concentration range 0.15-1.22 mM. In the clinically relevant concentration range the inhibition was small. At an isoflurane concentration of 0.31 mM (equivalent to 1 MAC), the peak N-type current was inhibited by 14 (1)%. The optical isomers of isoflurane were found to be equally potent at inhibiting currents through N-type channels. The inert gas anaesthetic xenon was found to have no measureable effect on N-type channels at a concentration of 3.4 mM (approximately 1 MAC). CONCLUSIONS: These results suggest that N-type calcium channels are not the targets mediating general anaesthesia with these two inhalation agents. PMID- 15778268 TI - Antagonism of neuromuscular blockade but not muscle relaxation affects depth of anaesthesia. AB - BACKGROUND: Conflicting effects of neuromuscular blocking drugs and anticholinesterases on depth of anaesthesia have been reported. Therefore we evaluated the effect of atracurium and neostigmine on bispectral index (BIS) and middle-latency auditory evoked potentials (AAI). METHODS: We studied 40 patients (ASA I-II) aged 18-69 yr. General anaesthesia consisted of propofol and remifentanil by target-controlled infusion and neuromuscular function was monitored by electromyography. When BIS reached stable values, patients were randomly assigned to one of two groups. Group 1 received atracurium 0.4 mg kg(-1) and, 5 min later, the same volume of NaCl 0.9%; group 2 received saline first and then atracurium. When the first twitch of a train of four reached 10% of control intensity, patients were again randomized: one group (N) received neostigmine 0.04 mg kg(-1) and glycopyrrolate 0.01 mg kg(-1), and the control group (G) received only glycopyrrolate. RESULTS: Injection of atracurium or NaCl 0.9% had no effect on BIS or AAI. After neostigmine-glycopyrrolate, BIS and AAI increased significantly (mean maximal change of BIS 7.1 [SD 7.5], P<0.001; mean maximal change of AAI 9.7 [10.5], P<0.001). When glycopyrrolate was injected alone BIS and AAI also increased (mean maximal change of BIS 2.2 [3.4], P=0.008; mean maximal change of AAI 3.5 [5.7], P=0.012), but this increase was significantly less than in group N (P=0.012 for BIS; P=0.027 for AAI). CONCLUSION: These data suggest that neostigmine alters the state of propofol-remifentanil anaesthesia and may enhance recovery. PMID- 15778269 TI - Bispectral index monitoring may not reliably indicate cerebral ischaemia during awake carotid endarterectomy. AB - BACKGROUND: Intraoperative ischaemia during carotid cross-clamping in patients undergoing carotid endarterectomy (CEA) is a major complication and prompt recognition of insufficient collateral blood supply is crucial. Electroencephalogram (EEG) is believed to be one of the useful forms of monitoring cerebrovascular insufficiency during CEA. The aim of this study was to evaluate the utility of bispectral index (BIS) monitoring, a processed EEG parameter, for the reliable detection of intraoperative cerebral ischaemia during awake CEA. METHODS: We monitored 52 patients continuously with the BIS monitor together with assessment of neurological function (contralateral upper and lower limb strength and the verbal component of the Glasgow Coma Scale for speech) in patients undergoing awake CEA. RESULTS: Overall mean BIS value in all patients was 96 (SD 2.9). In five patients who showed clinical evidence of cortical ischaemia during carotid cross-clamping, there was no change in the original range of BIS values throughout the procedure (96.7 [3.2]). In one patient BIS values decreased to 38 about 5 min after the incision and recovered within the next 10 min. The mean BIS value in the remaining 46 patients who did not develop clinical signs of ischaemia was 95.4 (2.6). Three cases are presented which demonstrate the inability of the BIS monitor to detect cerebral ischaemia. CONCLUSIONS: Lack of correlation of BIS with the signs of cerebral ischaemia during CEA makes it unreliable for detection of cerebrovascular insufficiency. We conclude that awake neurological testing is the preferred method of monitoring in these patients. PMID- 15778270 TI - Enzyme-selective effects of nitric oxide on affinity and maximum velocity of various rat cytochromes P450. AB - Nitric oxide (NO) has recently been shown to decrease cytochrome P450 (P450) enzyme activity rapidly (< or =30 min), concentration dependently, and enzyme selectively in the rat liver. Interestingly, among all the studied P450 enzymes, only CYP2D1 was not affected by NO donors. However, these studies were conducted using only a single concentration of the substrates, thus lacking information about the possible simultaneous changes in both maximum velocity (Vmax) and affinity (Km) of the enzymes. In the present study, we systematically evaluated the effects of NO on the enzyme kinetic parameters of marker substrates for a range of P450 enzymes, including 2D1. Livers were perfused (1 h) in the absence (control) or presence of two NO donors with different mechanisms of NO release. At the end of the perfusion, microsomes were prepared and used for kinetic analysis. Except for 2D1, NO reduced the Vmax of all the model reactions studied, although to a varying degree. However, the effects of NO donors on Km were more diverse. Whereas the Km values for testosterone 6beta-hydroxylation (3A2) and 16alpha-hydroxylation (2C11) significantly decreased, the values for chlorzoxazone 6-hydroxylation (2E1), dextromethorphan N-demethylation (3A2), and high affinity ethoxyresorufin O-dealkylation (1A1/2) significantly increased in the presence of NO donors. Furthermore, the Km values for the high-affinity component of dextromethorphan O-demethylation and benzyloxyresorufin O dealkylation remained unchanged. These results indicate that NO can potentially change both the Vmax and Km of various substrates selectively and confirm our previous findings that the activity of CYP2D1 is not affected by NO donors. PMID- 15778271 TI - Minipig cytochrome P450 2E1: comparison with human enzyme. AB - Cytochrome P450 2E1 was isolated from minipig liver microsomes. The protein has been cloned and the respective cDNA sequenced (GenBank Accession Number AY581116). Minipig CYP2E1 is two residues shorter than its human ortholog. The only difference between pig and minipig sequence is the presence of aspartic acid residue in position 346 contrary to valine in the pig enzyme. Minipig CYP2E1 was shown to be able to convert two prototypical substrates of human CYP2E1, chlorzoxazone and p-nitrophenol, to the respective metabolites. The experiments performed with both the liver microsomal fraction and reconstituted systems with human or minipig CYP2E1 confirmed the similarity of both enzymes. Inhibition with diethyldithiocarbamate gave comparable Ki values for minipig as well as for the human CYP2E1. The results indicate that the systems containing minipig CYP2E1 may be used to model the respective CYP2E1-catalyzed reactions of drug metabolism in humans. PMID- 15778272 TI - Impact of endotoxin-induced changes in P-glycoprotein expression on disposition of doxorubicin in mice. AB - P-glycoprotein (PGP) encoded by the Mdr1 gene mediates the excretion of drugs in organs such as the liver and kidney. Inflammation has been shown to suppress the expression and activity of PGP in rodent liver, thus potentially altering the pharmacokinetics of drugs that are substrates of PGP. Here we examined the effect of endotoxin (lipopolysaccharide; LPS)-induced inflammation on the disposition of the PGP substrate doxorubicin (DOX) in the mouse. Male CD-1 mice received 5 mg/kg LPS intraperitoneally. DOX (5 mg/kg) was administered intravenously 24 h after LPS treatment. The time course of DOX levels in plasma, urine, bile, and tissues was analyzed by high performance liquid chromatography. PGP protein and mRNA expression in liver and kidney was measured using Western blots and reverse transcriptase polymerase chain reaction. As compared to controls, LPS-treated mice exhibited a significant decrease (50%) in biliary clearance and 3-fold increased renal clearance of DOX. These changes were associated with strongly reduced PGP protein levels (30% controls, p < 0.05) in the liver and increased PGP levels in the kidney (140% controls, p < 0.05). Hepatic mRNA levels of all Mdr isoforms were reduced in LPS-treated mice, whereas renal Mdr1b levels were increased. In LPS-treated mice, we also measured an increased area under the plasma concentration-time curve and reduced systemic clearance of DOX, as well as a 2- to 5-fold increase in the urinary excretion of the doxorubicin and doxorubicinol aglycones. Our data suggest that endotoxin-induced inflammation in mice causes differential regulation of PGP in liver and kidney, thereby altering the clearance profile of DOX. PMID- 15778273 TI - Restoration of normal vascular relaxation mechanisms in cerebral arteries by chromosomal substitution in consomic SS.13BN rats. AB - This study sought to identify the mechanisms of vascular relaxation that are rescued in middle cerebral arteries (MCA) of SS.13BN consomic rats by substituting chromosome 13 containing the renin gene from Brown Norway (BN) rats into the Dahl salt-sensitive (SS) genetic background. Isolated MCA from SS rats exhibited an indomethacin-sensitive constriction in response to acetylcholine (ACh) and hypoxia. ACh-induced dilation was NO dependent and hypoxic dilations were cyclooxygenase (COX) dependent in BN and SS.13BN rats. In SS rats, hypoxic dilation was restored by indomethacin and abolished by inhibiting cytochrome P 450 epoxygenases, suggesting a role for epoxyeicosatrienoic acids. MCA from SS and SS.13BN rats constricted and MCA from BN rats dilated in response to the stable prostacyclin analog iloprost. MCA from SS.13BN and BN rats (but not SS rats) dilated in response to the prostaglandin E2 receptor agonist butaprost. Hypoxia increased prostacyclin release in cerebral arteries from all the strains, whereas thromboxane A2 production was reduced in BN rat vessels only. These data suggest that SS rats may be less sensitive to vasodilator prostaglandins and that normalization of renin-angiotensin system regulation causes a switch from production of COX-derived vasoconstrictor metabolites (in SS rats) toward NO dependent relaxation in response to ACh- and prostaglandin-dependent dilation in response to hypoxia in SS.13(BN) rats. PMID- 15778274 TI - Skeletal muscle reperfusion injury is enhanced in extracellular superoxide dismutase knockout mouse. AB - This study investigates the role of extracellular SOD (EC-SOD), the major extracellular antioxidant enzyme, in skeletal muscle ischemia and reperfusion (I/R) injury. Pedicled cremaster muscle flaps from homozygous EC-SOD knockout (EC SOD-/-) and wild-type (WT) mice were subjected to 4.5-h ischemia and 90-min reperfusion followed by functional and molecular analyses. Our results revealed that EC-SOD-/- mice showed significantly profound I/R injury compared with WT littermates. In particular, there was a delayed and incomplete recovery of arterial spasm and blood flow during reperfusion, and more severe acute inflammatory reaction and muscle damage were noted in EC-SOD-/- mice. After 90 min reperfusion, intracellular SOD [copper- and zinc-containing SOD (CuZn-SOD) and manganese-containing (Mn-SOD)] mRNA levels decreased similarly in both groups. EC-SOD mRNA levels increased in WT mice, whereas EC-SOD mRNA was undetectable, as expected, in EC-SOD-/- mice. In both groups of animals, CuZn-SOD protein levels decreased and Mn-SOD protein levels remained unchanged. EC-SOD protein levels decreased in WT mice. Histological analysis showed diffuse edema and inflammation around muscle fibers, which was more pronounced in EC-SOD-/- mice. In conclusion, our data suggest that EC-SOD plays an important role in the protection from skeletal muscle I/R injury caused by excessive generation of reactive oxygen species. PMID- 15778275 TI - A cis-9,trans-11-conjugated linoleic acid-rich oil reduces the outcome of atherogenic process in hyperlipidemic hamster. AB - Conjugated linoleic acid (CLA) mixtures demonstrated antiatherogenic properties in several animal models, including hamsters, but the mechanism of action of the main food-derived CLA isomer is unknown in this species. This study thus focused on cis-9,trans-11-CLA (rumenic acid), and its effect was compared with that of fish oil, which is known to influence several aspects of atherogenesis. Syrian hamsters were fed (for 12 wk) diets containing 20% (wt/wt) butter fat (B diet) or the same diet augmented with either 1% (wt/wt) of a cis-9,trans-11-CLA-rich oil (BR diet) or 1% (wt/wt) fish oil (BF diet). The BR diet induced the lowest aortic lipid deposition (from -30% to -45%) among the butter oil-fed hamsters. In this group, plasma also displayed a reduced non-HDL-to-HDL-cholesterol ratio (21% less than in the butter oil group) and inflammatory serum amyloid A levels (70-80%) and an improvement of anti-oxidized LDL paraoxonase activity (all P < 0.05). Compared with the B group, the beneficial effects of the BR diet could be further explained in part by preventing the high VCAM-1 expression rate, increasing (30%) ATP-binding cassette subfamily A1 expression in the aorta, and downregulating expression of inflammatory-related genes (TNF-alpha, IL-1beta, and cyclooxygenase 2, 2- to 2.8-fold, P < 0.05). This effect was partly associated with an activation of peroxisome proliferator-activating receptor (PPAR)/liver X receptor (LXR)-alpha signaling cascade. Interestingly, activation of PPAR/LXR-alpha signaling was not observed in hamsters fed the BF diet, in which the early signs of atherogenesis were increased. In conclusion, this study demonstrated that milk fat-rich cis-9,trans-11-CLA reduces the atherogenic process in hyperlipidemic hamsters. PMID- 15778276 TI - Cardiac-specific attenuation of natriuretic peptide A receptor activity accentuates adverse cardiac remodeling and mortality in response to pressure overload. AB - Atrial (ANP) and brain (BNP) natriuretic peptides are hormones of myocardial cell origin. These hormones bind to the natriuretic peptide A receptor (NPRA) throughout the body, stimulating cGMP production and playing a key role in blood pressure control. Because NPRA receptors are present on cardiomyocytes, we hypothesized that natriuretic peptides may have direct autocrine or paracrine effects on cardiomyocytes or adjacent cardiac cells. Because both natriuretic peptides and NPRA gene expression are upregulated in states of pressure overload, we speculated that the effects of the natriuretic peptides on cardiac structure and function would be most apparent after pressure overload. To attenuate cardiomyocyte NPRA activity, transgenic mice with cardiac specific expression of a dominant-negative (DN-NPRA) mutation (HCAT D 893A) in the NPRA receptor were created. Cardiac structure and function were assessed (avertin anesthesia) in the absence and presence of pressure overload produced by suprarenal aortic banding. In the absence of pressure overload, basal and BNP-stimulated guanylyl cyclase activity assessed in cardiac membrane fractions was reduced. However, systolic blood pressure, myocardial cGMP, log plasma ANP levels, and ventricular structure and function were similar in wild-type (WT-NPRA) and DN-NPRA mice. In the presence of pressure overload, myocardial cGMP levels were reduced, and ventricular hypertrophy, fibrosis, filling pressures, and mortality were increased in DN-NPRA compared with WT-NPRA mice. In addition to their hormonal effects, endogenous natriuretic peptides exert physiologically relevant autocrine and paracrine effects via cardiomyocyte NPRA receptors to modulate cardiac hypertrophy and fibrosis in response to pressure overload. PMID- 15778277 TI - gp91phox-containing NAD(P)H oxidase mediates attenuation of nitric oxide dependent control of myocardial oxygen consumption by ANG II. AB - We have previously reported that ANG II stimulation increased superoxide anion (O2-) through the activation of NAD(P)H oxidase and inhibited nitric oxide (NO) dependent control of myocardial oxygen consumption (MVo2) by scavenging NO. Our objective was to investigate the role of NAD(P)H oxidase, especially the gp91phox subunit, in the NO-dependent control of MVo2. MVo2 in mice with defects in the expression of gp91phox [gp91(phox)(-/-)] was measured with a Clark-type oxygen electrode. Baseline MVo2 was not significantly different between wild-type (WT) and gp91(phox)(-/-) mice. Stimulation of NO production by bradykinin (BK) induced significant decreases in MVo2 in WT mice. BK-induced reduction in MVo2 was enhanced in gp91(phox)(-/-) mice. BK-induced reduction in MVo2 in WT mice was attenuated by 10(-8) mol/l ANG II, which was restored by coincubation with Tiron or apocynin. In contrast to WT mice, BK-induced reduction in MVo2 in gp91(phox)( /-) mice was not altered by ANG II. There was a decrease in lucigenin (5 x 10(-6) mol/l)-detectable O2- in gp91(phox)(-/-) mice compared with WT mice. ANG II resulted in significant increases in O2- production in WT mice, which was inhibited by coincubation with Tiron or apocynin. However, ANG II had no effect on O2- production in gp91(phox)(-/-) mice. Histological examination showed that the development of abscesses and/or the invasion of inflammatory cells occurred in lungs and livers but not in hearts and kidneys from gp91(phox)(-/-) mice. These results indicate that the gp91(phox) subunit of NAD(P)H oxidase mediates O2 production through the activation of NAD(P)H oxidase and attenuation of NO dependent control of MVo2 by ANG II. PMID- 15778278 TI - Dietary isoflavones during pregnancy and lactation provide cardioprotection to offspring rats in adulthood. AB - In adult rats, elongation of cardiac myocytes (CMs) correlates with dilatation (and sometimes dysfunction) of cardiac ventricles. Although sex steroids may constitute one possible factor that affects the dimensions of CMs, studies on their effects in rodents is complicated by the fact that most commercial soy based diets also contain abundant phytoestrogens. We report that feeding Wistar Kyoto rat dams during gestation and lactation with a phytoestrogen-rich soy-based diet caused the CMs of their adult offspring to be shorter than in counterparts originating from mothers fed with a phytoestrogen-free casein-based diet. The soy based diet had no such effects when given to rats after 6 wk of age, and its effects were replicated when supplementing the maternal casein-based diet with the isoflavones daidzein and genistein (the most abundant phytoestrogens in soy based diets). In contrast to rats whose mothers had been fed with a soy-based diet, the hearts of adult rats raised with a casein-based diet only featured dilated eccentric hypertrophy and progressed toward congestive heart failure when further challenged. Thus the presence of isoflavones in the maternal diet provides cardioprotection to the hearts of their offspring during adulthood. PMID- 15778279 TI - Menstrual cycle and sex affect hemodynamic responses to combined orthostatic and heat stress. AB - Women have decreased orthostatic tolerance compared with men, and anecdotal evidence suggests women are more susceptible to orthostatic intolerance in warm environments. Because estrogen and progesterone affect numerous physiological variables that may alter orthostatic tolerance, the purpose of our study was to compare orthostatic tolerance across the menstrual cycle phases in women during combined orthostatic and heat stress and to compare these data with those of men. Eight normally menstruating women and eight males (22 +/- 4.0 and 23 +/- 3.5 yr, respectively) completed the protocol. Women were studied during their early follicular (EF), ovulatory (OV), and midluteal (ML) phases. Men were studied twice within 2-4 wk. Heart rate, cardiac output, blood pressure, core temperature (T(c)), and cutaneous vascular conductance (CVC) were measured during three head up tilt tests, consisting of two tilts in the thermoneutral condition and one tilt after a 0.5 degrees C rise in T(c). There was no difference in orthostatic tolerance across the menstrual cycle phases, despite higher CVC in the ML phase after heating (EF, 42.3 +/- 4.8; OV, 40.1 +/- 3.7; ML, 57.5 +/- 4.5; P < 0.05). Orthostatic tolerance in the heat was greater in men than women (P < 0.05). These data suggest that although many physiological variables associated with blood pressure regulation fluctuate during the menstrual cycle, orthostatic tolerance in the heat remains unchanged. Additionally, our data support a clear sex difference in orthostatic tolerance and extend upon previous data to show that the sex difference in the heat is not attributable to fluctuating hormone profiles during the menstrual cycle. PMID- 15778280 TI - Sphingosine 1-phosphate prevents platelet-activating factor-induced increase in hydraulic conductivity in rat mesenteric venules: pertussis toxin sensitive. AB - Sphingosine 1-phosphate (S1P) is a biologically active lipid. In vitro, S1P tightens the endothelial barrier, as assessed by a rapid increase in electrical resistance and a decrease in solute permeability. We hypothesized that this activity of S1P would also occur in vivo. Hydraulic conductivity (Lp), an assessment of endothelial barrier function, was measured in individually perfused venules in rat mesenteries. S1P (1 microM) decreased basal Lp by 63% when basal Lp was between 3.6 and 4.1 x 10(-7) cm x s(-1) x cmH2O(-1) but showed no effect when basal Lp was below 2 x 10(-7) cm x s(-1) x cmH2O(-1). Under either condition, S1P blocked the sixfold increase in Lp induced by platelet-activating factor (PAF, 10 nM). Perfusion of venules with pertussis toxin (0.1 microg/ml), a specific inhibitor of the inhibitory G protein, Gi, for 3 h did not affect basal Lp or the increased Lp induced by PAF. Pertussis toxin, however, significantly attenuated the inhibitory action of S1P on the PAF-induced increase in Lp, indicating the involvement of the Gi protein. Measurement of endothelial cytoplasmic Ca2+ concentration ([Ca2+]i) in venules loaded with fura-2 AM showed that S1P alone transiently increased basal endothelial [Ca2+]i (from 89 nM to 193 nM) but had no effect on the magnitude and time course of the PAF-induced increase in endothelial [Ca2+]i. These results indicate that S1P functions in vivo to prevent the PAF-induced increase in microvessel permeability. The inhibitory action of S1P involves the pertussis toxin-sensitive Gi protein and is not mediated by prevention of the PAF-induced increase in endothelial [Ca2+]i. PMID- 15778281 TI - A B56 regulatory subunit of protein phosphatase 2A localizes to nuclear speckles in cardiomyocytes. AB - Protein phosphatase 2A (PP2A) is widely distributed in heart tissues, yet its precise cellular functions are poorly understood. This study is based on the notion that PP2A action is governed by interactions of the core enzyme with B targeting/regulatory subunits. The subcellular localizations of two B subunits, B56alpha and B56gamma1, were assessed using adenovirus-driven expression of epitope-tagged (hemagglutinin, HA) in cultured neonatal and adult rat ventricular myocytes. Confocal imaging revealed that HA-B56alpha was excluded from the nucleus and decorated striated structures, whereas HA-B56gamma1 was principally found in the nucleus. Precise immunolabeling studies showed that B56gamma1 was concentrated in intranuclear structures known as nuclear speckles, macromolecular structures that accumulate transcription and splicing factors. Western blot analyses revealed that overexpression of either B subunit had no effect on the levels of other PP2A subunits in cultured neonatal cardiac cells. However, overexpression of only B56gamma1 increased whole cell PP2A activity by 40% when measured in cell extracts. Finally, B56gamma1 did not alter global gene expression or expression of hypertrophic gene markers such as alpha-skeletal actin. However, morphometric analyses of confocal images revealed that B56gamma1 alters the dynamic assembly/disassembly process of nuclear speckles in heart cells. These studies provide new insight into mechanisms of PP2A targeting in the subnuclear architecture in cardiomyocytes and into the role of this phosphatase in nuclear signaling. PMID- 15778282 TI - Sympathetic activation restrains endothelium-mediated muscle vasodilatation in heart failure patients. AB - Although the vasodilatory response during mental stress is blunted in heart failure (HF), the mechanisms underlying this phenomenon are not fully understood. We tested the hypothesis that sympathetic activity limits the endothelium dependent vasodilatation during mental stress in chronic HF patients. Twenty-one HF patients (age 45 +/- 2 yr, functional classes III and IV, New York Heart Association) and 22 age-matched normal controls (NC; age 42 +/- 2 yr, P = 0.13) were studied at rest and during 4 min of Stroop color-word test with brachial intra-arterial saline, acetylcholine (endothelium dependent), phentolamine (alpha blocker), and phentolamine plus acetylcholine infusion. Forearm blood flow was measured by venous occlusion plethysmography. Baseline forearm vascular conductance (FVC) was significantly lower in HF patients (2.18 +/- 0.12 vs. 3.66 +/- 0.22 units, P = 0.001). During mental stress with saline, the changes in FVC were significantly blunted in HF patients compared with NC (0.92 +/- 0.20 vs. 2.13 +/- 0.39 units, P = 0.001). In HF, the vasodilatation with acetylcholine was similar to saline control and significantly lower than in NC. In HF patients, phentolamine significantly increased FVC responses (1.16 +/- 0.20 vs. 2.09 +/- 0.29 units, P = 0.001), and the difference between HF patients and NC tended to decrease (2.09 +/- 0.29 vs. 3.61 +/- 0.74 units, P = 0.052). The vasodilatation with phentolamine plus acetylcholine was similar between HF and NC (4.23 +/- 0.73 vs. 4.76 +/- 1.03 units, P = 0.84). In conclusion, sympathetic activation mediates the blunted muscle endothelium-mediated vasodilatation during mental stress in HF patients. PMID- 15778283 TI - MRI-based finite-element analysis of left ventricular aneurysm. AB - Tagged MRI and finite-element (FE) analysis are valuable tools in analyzing cardiac mechanics. To determine systolic material parameters in three-dimensional stress-strain relationships, we used tagged MRI to validate FE models of left ventricular (LV) aneurysm. Five sheep underwent anteroapical myocardial infarction (25% of LV mass) and 22 wk later underwent tagged MRI. Asymmetric FE models of the LV were formed to in vivo geometry from MRI and included aneurysm material properties measured with biaxial stretching, LV pressure measurements, and myofiber helix angles measured with diffusion tensor MRI. Systolic material parameters were determined that enabled FE models to reproduce midwall, systolic myocardial strains from tagged MRI (630 +/- 187 strain comparisons/animal). When contractile stress equal to 40% of the myofiber stress was added transverse to the muscle fiber, myocardial strain agreement improved by 27% between FE model predictions and experimental measurements (RMS error decreased from 0.074 +/- 0.016 to 0.054 +/- 0.011, P < 0.05). In infarct border zone (BZ), end-systolic midwall stress was elevated in both fiber (24.2 +/- 2.7 to 29.9 +/- 2.4 kPa, P < 0.01) and cross-fiber (5.5 +/- 0.7 to 11.7 +/- 1.3 kPa, P = 0.02) directions relative to noninfarct regions. Contrary to previous hypotheses but consistent with biaxial stretching experiments, active cross-fiber stress development is an integral part of LV systole; FE analysis with only uniaxial contracting stress is insufficient. Stress calculations from these validated models show 24% increase in fiber stress and 115% increase in cross-fiber stress at the BZ relative to remote regions, which may contribute to LV remodeling. PMID- 15778284 TI - Activation of regenerating gene Reg in rat and human hearts in response to acute stress. AB - Recently, the regenerating gene (Reg) has been documented to play an important role in various regenerating tissues, but it is unknown whether the Reg gene could be activated in the heart. The aim of this study was to reveal the transcriptional activation of Reg in the heart in response to heart stress. We first found REG-1 protein expression in human hearts obtained from autopsied patients who died of myocardial infarction. REG protein was immunohistochemically stained in a fine granular pattern in the cytoplasm of cardiomyocytes. To demonstrate the activation profiles of Reg gene expression in the heart, we quantified the levels of Reg-1 mRNA in rat hearts after coronary artery ligation using real-time RT-PCR. Transient Reg-1 mRNA activation, peaking at 12 h after coronary ligation, was observed mainly in the atria, which was sevenfold higher compared with hearts with pressure overload due to aortic constriction. In contrast, Reg receptor mRNA was expressed intensely in damaged ventricles. Furthermore, Western blot analysis showed the corresponding pattern of Reg protein secretion into the serum after loading, and circulating levels of the protein after myocardial infarction were higher than those after aortic constriction. In conclusion, our results demonstrate for the first time the presence of the Reg/Reg receptor system in damaged hearts. In view of emerging evidence of Reg for tissue regeneration in a variety of tissues/organs, it is proposed that the damaged heart may be a target for Reg action and that Reg may protect against acute heart stress. PMID- 15778285 TI - Heterogeneous vasodilator responses of human limbs: influence of age and habitual endurance training. AB - Forearm endothelium-dependent vasodilation is impaired with age in sedentary, but not endurance-trained, men. The purpose of this investigation was to determine whether these age- and physical activity-related differences in endothelium dependent vasodilation also occur in the leg. Brachial and common femoral arterial blood flow were measured with Doppler ultrasound during increasing doses of acetylcholine (1, 4, and 16 microg.100 ml limb tissue(-1).min(-1)), substance P (8, 31, and 125 pg.100 ml limb tissue(-1).min(-1)), and sodium nitroprusside (0.063, 0.25, and 1 microg.100 ml limb tissue(-1).min(-1)) in 23 healthy men (8 younger sedentary, 8 older sedentary, and 7 older endurance trained). Increases in forearm blood flow to the highest dose of acetylcholine and sodium nitroprusside were smaller (P < 0.05) in older sedentary (841 +/- 142%, 428 +/- 74%) compared with younger sedentary (1,519 +/- 256%, 925 +/- 163%) subjects. Similarly, increases in forearm blood flow to sodium nitroprusside (1 microg.100 ml limb tissue(-1).min(-1)) were smaller (P < 0.05) in older endurance-trained (505 +/- 110%) compared with younger sedentary (925 +/- 163%) subjects. In contrast, no differences in leg blood flow responses to intra-arterial infusions of acetylcholine, substance P, or sodium nitroprusside were noted between subject groups. These results demonstrate that 1) acetylcholine- and sodium nitroprusside induced vasodilation are attenuated in the forearm vasculature and preserved in the leg vasculature of older sedentary subjects and 2) sodium nitroprusside induced vasodilation remains attenuated in the forearm vasculature of healthy older endurance-trained men but preserved in the leg vasculature of these men. PMID- 15778286 TI - Simulated microgravity produces attenuated baroreflex-mediated pressor, chronotropic, and inotropic responses in mice. AB - Whether myocardial contractile impairment contributes to orthostatic intolerance (OI) is controversial. Accordingly, we used transient bilateral carotid occlusion (TBCO) to compare the in vivo pressor, chronotropic, and inotropic responses (parts 1 and 2) to open-loop selective carotid baroreceptor unloading in anesthetized mice. In part 3, in vitro myocyte responses to isoproterenol in mice exposed to hindlimb unweighting (HLU) for approximately 2 wk were determined. Heart rate (HR) and mean arterial pressure (MAP) responses to TBCO were measured. In control mice, TBCO increased HR (15 +/- 2 beats/min, P < 0.05) and MAP (17 +/- 2 mmHg, P < 0.05). These responses were markedly potentiated in denervated control (DC) mice, in which the aortic depressor nerve and sympathetic trunk were sectioned before measurement. Baroreflex responses to TBCO were eliminated by blockade with hexamethonium bromide (10 microg/kg). In HLU (denervated) mice, HR and MAP responses were reduced approximately 70% compared with DC mice. In part 2, myocardial contractile responses to TBCO were measured with a left ventricular micromanometer-conductance catheter. TBCO in DC mice increased the slope of the end-systolic pressure-volume relation (end-systolic elastance) by 86 +/- 13%. This inotropic response was attenuated (14 +/- 10%, P < 0.005) after HLU. In part 3, contractile responses to isoproterenol were impaired in myocytes isolated from HLU mice. In conclusion, selective carotid baroreceptor unloading stimulates HR, blood pressure, and myocardial contractility, and HLU attenuates each response. These findings have important implications for the management of OI in astronauts, the elderly, and individuals subjected to prolonged bed rest. PMID- 15778287 TI - Role of endogenous nitric oxide in endotoxin-induced alteration of hypoxic pulmonary vasoconstriction in mice. AB - Pulmonary vasoconstriction in response to alveolar hypoxia (HPV) is frequently impaired in patients with sepsis or acute respiratory distress syndrome or in animal models of endotoxemia. Pulmonary vasodilation due to overproduction of nitric oxide (NO) by NO synthase 2 (NOS2) may be responsible for this impaired HPV after administration of endotoxin (LPS). We investigated the effects of acute nonspecific (N(G)-nitro-L-arginine methyl ester, L-NAME) and NOS2-specific [L-N6 (1-iminoethyl)lysine, L-NIL] NOS inhibition and congenital deficiency of NOS2 on impaired HPV during endotoxemia. The pulmonary vasoconstrictor response and pulmonary vascular pressure-flow (P-Q) relationship during normoxia and hypoxia were studied in isolated, perfused, and ventilated lungs from LPS-pretreated and untreated wild-type and NOS2-deficient mice with and without L-NAME or L-NIL added to the perfusate. Compared with lungs from untreated mice, lungs from LPS challenged wild-type mice constricted less in response to hypoxia (69 +/- 17 vs. 3 +/- 7%, respectively, P < 0.001). Perfusion with L-NAME or L-NIL restored this blunted HPV response only in part. In contrast, LPS administration did not impair the vasoconstrictor response to hypoxia in NOS2-deficient mice. Analysis of the pulmonary vascular P-Q relationship suggested that the HPV response may consist of different components that are specifically NOS isoform modulated in untreated and LPS-treated mice. These results demonstrate in a murine model of endotoxemia that NOS2-derived NO production is critical for LPS-mediated development of impaired HPV. Furthermore, impaired HPV during endotoxemia may be at least in part mediated by mechanisms other than simply pulmonary vasodilation by NOS2 derived NO overproduction. PMID- 15778288 TI - Critical contribution of CD80 and CD86 to induction of anterior chamber associated immune deviation. AB - Intraocular inoculation of antigens induces anterior chamber-associated immune deviation (ACAID), which is mediated by development of regulatory T cells in response to antigen-presenting cells (APC) pre-conditioned by intraocular transforming growth factor-beta (TGF-beta). In this study, we examined the involvement of T-cell co-stimulatory molecules in this process. To mimic the intraocular APC, thioglycollate-elicited peritoneal exudate cells (PEC) were pre treated with TGF-beta in vitro. Expression of CD80, CD86, OX40 ligand (OX40L) and CD70 was analyzed by flow cytometry. Contribution of these molecules to co stimulatory activity of TGF-beta-treated PEC on antigen-stimulated T-cell proliferation and cytokine production was determined by inhibition with blocking antibodies in vitro. Contribution of CD80 and CD86 to induction of ACAID was determined by the administration of blocking antibodies at intraocular antigen inoculation in vivo. TGF-beta-treated PEC expressed CD80 and CD86 but not OX40L or CD70. Antigen-stimulated T cells proliferated and produced IL-10, but not IFN gamma, in response to co-stimulation by TGF-beta-treated PEC, which was abrogated by blocking antibodies against CD80 and CD86. Induction of regulatory cells mediating ACAID was abolished by in vivo blockade of CD80 and CD86. The present results indicated that CD80 and CD86 play a critical role in induction of ACAID, possibly by co-stimulating expansion and IL-10 production of regulatory T cells in response to TGF-beta-conditioned APC. PMID- 15778289 TI - Highly efficient antigen targeting to M-DC8+ dendritic cells via FcgammaRIII/CD16 specific antibody conjugates. AB - Conjugates of peptide antigens with antibodies specifically recognizing surface molecules on dendritic cells (DC) represent an attractive approach to target antigens to antigen-presenting cells (APC) for the induction of specific T cell responses. The present study evaluates the potential of M-DC8(+) DC, a sub population of professional APC in the blood, for an antibody-based vaccination strategy. We prepared, by chemical cross-linking, conjugates of peptide model antigens with antibodies directed against different cell surface molecules of DC. Antigen-peptide conjugates using an anti-CD16 (FcgammaRIII) antibody were most potent in inducing in vitro activation of a specific CD4(+) T cell response. They were at least 300 times more efficient than two other antibody-antigen conjugates and approximately 500 times more efficient than unconjugated antigen peptides. Our data demonstrate that specific antigen targeting via CD16 on M-DC8(+) DC is a promising vaccination approach for the efficient induction of specific CD4(+) T cell responses ex vivo, and perhaps in vivo. PMID- 15778290 TI - Anti-CD3 induces bi-phasic apoptosis in murine intestinal epithelial cells: possible involvement of the Fas/Fas ligand system in different T cell compartments. AB - Recent studies have suggested that Fas-mediated apoptosis is involved in the pathogenesis of intestinal injury. In this study, we determined the role of Fas/Fas ligand (FasL) interactions in different T cell compartments using a murine model of small intestinal injury. An intraperitoneal injection of 145-2C11 (anti-CD3) antibody into C3H/HeN, BALB/c and MRL mice induced mucosal flattening and rapid, bi-phasic intestinal epithelial cell (IEC) apoptosis, which was detected by conventional light and electron microscopy and by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. In the first, early phase, villous apoptosis was observed up to 4 h after injection, and in the second, later phase, apoptotic crypt cells gradually accumulated for up to 24 h. The early and later phases of apoptosis were reduced in lpr/lpr and nude mice compared with those in control strains. In addition, the kinetics of Fas-mediated killer activity induced by the antibody injection were different between intestinal intraepithelial lymphocytes (IEL) and splenocytes (SPL) and seemed to correlate with the bi-phasic occurrence of the apoptosis. Finally, the transfer of intestinal IEL from euthymic to nude mice induced both phases of apoptosis, whereas SPL induced the second phase's crypt apoptosis only by the antibody injection. Together, these results suggest the involvement of Fas-mediated killer activity of thymus-derived T cells in different compartments. Namely, T cell populations in different compartments are differentially involved in the induction of IEC apoptosis and contribute to the complex pathogenesis of immune mediated intestinal injury in which Fas/FasL interactions may play a critical role. PMID- 15778291 TI - Thyroid hormone can increase estrogen-mediated transcription from a consensus estrogen response element in neuroblastoma cells. AB - Thyroid hormones (T) and estrogens (E) are nuclear receptor ligands with at least two molecular mechanisms of action: (i) relatively slow genomic effects, such as the regulation of transcription by cognate T receptors (TR) and E receptors (ER); and (ii) relatively rapid nongenomic effects, such as kinase activation and calcium release initiated at the membrane by putative membrane receptors. Genomic and nongenomic effects were thought to be disparate and independent. However, in a previous study using a two-pulse paradigm in neuroblastoma cells, we showed that E acting at the membrane could potentiate transcription from an E-driven reporter gene in the nucleus. Because both T and E can have important effects on mood and cognition, it is possible that the two hormones can act synergistically. In this study, we demonstrate that early actions of T via TRalpha1 and TRbeta1 can potentiate E-mediated transcription (genomic effects) from a consensus E response element (ERE)-driven reporter gene in transiently transfected neuroblastoma cells. Such potentiation was reduced by inhibition of mitogen activated protein kinase. Using phosphomutants of ERalpha, we also show that probable mitogen-activated protein kinase phosphorylation sites on the ERalpha, the serines at position 167 and 118, are important in TRbeta1-mediated potentiation of ERalpha-induced transactivation. We suggest that crosstalk between T and E includes potential interactions through both nuclear and membrane initiated molecular mechanisms of hormone signaling. PMID- 15778292 TI - An initial strategy for the systematic identification of functional elements in the human genome by low-redundancy comparative sequencing. AB - With the recent completion of a high-quality sequence of the human genome, the challenge is now to understand the functional elements that it encodes. Comparative genomic analysis offers a powerful approach for finding such elements by identifying sequences that have been highly conserved during evolution. Here, we propose an initial strategy for detecting such regions by generating low redundancy sequence from a collection of 16 eutherian mammals, beyond the 7 for which genome sequence data are already available. We show that such sequence can be accurately aligned to the human genome and used to identify most of the highly conserved regions. Although not a long-term substitute for generating high quality genomic sequences from many mammalian species, this strategy represents a practical initial approach for rapidly annotating the most evolutionarily conserved sequences in the human genome, providing a key resource for the systematic study of human genome function. PMID- 15778294 TI - Sonic hedgehog and retinoic acid synergistically promote sensory fate specification from bone marrow-derived pluripotent stem cells. AB - Recent studies demonstrated that stromal cells isolated from adult bone marrow have the competence of differentiating into neuronal cells in vitro and in vivo. However, the capacity of marrow stromal cells or mesenchymal stem cells (MSCs) to differentiate into diverse neuronal cell populations and the identity of molecular factors that confer marrow stromal cells with the competence of a neuronal subtype have yet to be elucidated. Here, we show that Sonic hedgehog (Shh) and retinoic acid (RA), signaling molecules secreted from tissues in the vicinity of peripheral sensory ganglia during embryogenesis, exert synergistic effects on neural-competent MSCs to express a comprehensive set of glutamatergic sensory neuron markers. Application of Shh or RA alone had little or no effect on the expression of these neuronal subtype markers. In addition, incubation of MSCs with embryonic hindbrain/somite/otocyst conditioned medium or prenatal cochlea explants promoted up-regulation of additional sensory neuron markers and process outgrowth. These results identify Shh and RA as sensory competence factors for adult pluripotent cells and establish the importance of interactions between adult pluripotent cells and the host microenvironment in neuronal subtype specification. PMID- 15778293 TI - Ku70 acetylation mediates neuroblastoma cell death induced by histone deacetylase inhibitors. AB - Histone deacetylase inhibitors (HDACIs) are therapeutic drugs that inhibit deacetylase activity, thereby increasing acetylation of many proteins, including histones. HDACIs have antineoplastic effects in preclinical and clinical trials and are being considered for cancers with unmet therapeutic need, including neuroblastoma (NB). Uncertainty of how HDACI-induced protein acetylation leads to cell death, however, makes it difficult to determine which tumors are likely to be responsive to these agents. Here, we show that NB cells are sensitive to HDACIs, and that the mechanism by which HDACIs induce apoptosis involves Bax. In these cells, Bax associates with cytoplasmic Ku70, a protein that typically increases chemotherapy resistance. Our data show that in NB cells Ku70 binds to Bax in an acetylation-sensitive manner. Upon HDACI treatment, acetylated Ku70 releases Bax, allowing it to translocate to mitochondria and trigger cytochrome c release, leading to caspase-dependent death. This study shows that Ku70 is an important Bax-binding protein, and that this interaction can be therapeutically regulated in NB cells. Whereas the Bax-binding ability of Ku70 allows it to block apoptosis in response to certain agents, it is also a molecular target for the action of HDACIs, and in this context, a mediator of NB cell death. PMID- 15778295 TI - Melt analysis of process cheese spread or product using a rapid visco analyzer. AB - The objective of this study was to evaluate a rapid visco analyzer (RVA) method for measuring the melting characteristics of process cheese spread or product. The melt properties of 32 commercial process cheese spread and process cheese product samples from 4 manufacturers were analyzed with the RVA, tube melt test, texture profile analysis (TPA) hardness, and dynamic stress rheometry (DSR). For the RVA melt test, a 15-g disc of cheese was packed into the RVA canister and subjected to a heating, holding, and cooling profile during continuous mixing. During the test, the apparent viscosity was continuously measured and several data points (melt time, hot viscosity, time at 5000 cP during cooling, and solidification time) were collected from the viscosity vs. time curve. There was a high correlation (R2= 0.91) between the DSR melt temperature and the tube melt test. There was also a high correlation between the RVA melt time and the DSR melt temperature or the tube melt test (R2= 0.84 and 0.74, respectively). The RVA hot viscosity had a low correlation (R2< 0.44) with the DSR melt temperature and the tube melt test but had a high correlation (R2= 0.74) with DSR G'' at 85 degrees C. The results of this study indicate that RVA melt analysis of process cheese spread/product is correlated with the results from other melt tests and is capable of measuring the melt properties quantified by other methods. The RVA melt test may also provide additional information on the melt characteristics of process cheese spread/product not measured in other tests. PMID- 15778296 TI - Evaluation of bitterness in Ragusano cheese. AB - The appearance of undesirable bitter taste in Ragusano cheese was investigated by comparing the composition of 9 bitter cheeses with that of 9 reference cheeses of good quality by means of chemical, electrophoretic, and chromatographic analyses. Rates of proteolysis were significantly affected in cheeses of different quality. Primary proteolysis, as measured by pH 4.6-soluble N, was significantly greater in bitter cheeses compared with reference samples. Urea-PAGE profiles showed an almost complete breakdown of caseins in bitter cheeses and the further degradation of primary peptides into smaller compounds not detectable by this technique. Cheeses with defects had significantly lower levels of secondary proteolysis as reflected by the percentage of pH 4.6-soluble N soluble in 12% trichloroacetic acid and the amounts of total free amino acids. Peptides separated by reversed phase-HPLC revealed that the large and significant differences in peptide profiles of the soluble fractions between bitter and reference cheeses were mainly due to a much higher proportion of hydrophobic peptides in the former. The occurrence of bitterness in Ragusano cheese was therefore attributable to unbalanced levels of proteolysis and peptidolysis. Extensive degradation of caseins and primary peptides by activities of proteases produced large amounts of small- and medium-sized hydrophobic peptides that were not adequately removed by peptidases of microflora and therefore accumulated in cheese potentially contributing to its bitter taste. The presence of these compounds in bitter cheeses was related to high salt-in-moisture and low moisture contents that limited the enzymatic activities of microflora important in secondary proteolysis. Combining salt-in-moisture and the ratio of hydrophobic-to hydrophilic soluble peptides resulted in the best logistic partial least squares regression model predicting cheese quality. Although bitterness is known to be rarely encountered in cheese at salt-in-moisture levels >5.0, all of the bitter cheeses analyzed in this study had salt-in-moisture levels much greater than this value. According to the logistic model, a risk of bitterness development may exist for cheeses with a midrange (5 to 10%) salt-in-moisture content but with an inadequate level of secondary proteolysis. PMID- 15778297 TI - Effect of heating and processing methods of milk and dairy products on conjugated linoleic acid and trans fatty Acid isomer content. AB - The conventional heating methods of milk did not cause any significant increase in the trans isomer content, with the exception of milk heated at 63 +/- 1.0 degrees C for 30 min and milk microwaved for 5 min, which were significantly increased by 19 and 31%, respectively. The chemical changes of lipids were generally accelerated with the severity of the heat treatment and duration of storage. The conjugated linoleic acid content of cheese heated in a microwave oven for 5 min decreased by 21%, and microwave heating for 10 min caused a decrease of 53% compared with that of freshly boiled cheese. PMID- 15778298 TI - Seasonal variation in conjugated linoleic acid and vaccenic acid in milk fat of sheep and its transfer to cheese and ricotta. AB - The seasonal variation in conjugated linoleic acid (CLA) and vaccenic acid (VA) concentrations in sheep dairy products and the extent of their transfer from milk fat to cheese and ricotta fat were investigated. Samples were collected from 2 sheep milk processing plants in North Sardinia (Italy) every 2 wk from March through June. Concentrations of fatty acids (FA) in fresh cheese and ricotta fat were primarily dependent on the fatty acid content of the unprocessed raw milk. The content of c9,t11-CLA averaged 1.73, 1.69, and 1.75 mg/100 mg of FA methyl esters (FAME), respectively, for milk, cheese, and ricotta, and differed significantly between cheese and ricotta. The content of VA averaged 3.40, 3.33, and 3.43 mg/100 mg of FAME, respectively for milk, cheese, and ricotta. The FA composition of dairy products was markedly affected by period of sampling: the mean c9,t11-CLA and VA concentration decreased from March (2.20 and 4.52 mg/100 mg of FAME) to June (1.14 and 1.76 mg/100 mg of FAME) in all dairy products. No differences in c9,t11-CLA and VA concentration of dairy products were observed between the 2 dairy companies obtaining milk from the same geographical origin. The seasonal changes in CLA and VA in milk fat were probably related to changes in pasture quality. PMID- 15778299 TI - Use of ultrasound for characterizing dairy products. AB - It has been known for a long time that acoustic measurements offer some unique features for characterizing liquid food products in their intact state, without any preparation or destruction of the product sample. Acoustic characterization can yield information about fat content, droplet size distribution, and kinetics of product variation with time. Furthermore, acoustic methods are very attractive for on-line process control. This paper addresses several questions: Why does ultrasound attenuate when propagating through a heterogeneous system? What properties of dairy products can be extracted from such ultrasound measurements? Which measurement is better for product characterization: attenuation or sound speed? What measurement precision is required to adequately characterize product properties? What frequency range is of most value for determining these product properties?It is possible to provide answers to many of these questions using experimental data, thereby avoiding, for the moment, any complex mathematical analysis. We present several applications of acoustic spectroscopy for characterizing dairy products, including characterization of the fat content in a wide variety of dairy products; calculation of the fat droplet size distribution in milk without dilution; and calculation of water droplet size in butter, without dilution or melting. PMID- 15778300 TI - Antagonistic action of Lactobacillus delbrueckii ssp. lactis RM2-5 toward spoilage organisms in cottage cheese. AB - Cells of Lactobacillus delbrueckii ssp. lactis RM2-5 were added to cottage cheese stored at 7 degrees C in different amounts to determine if they would inhibit the growth of Pseudomonas fluorescens, also inoculated into the cheese samples. In addition, experiments were conducted in which no spoilage organisms were added to determine the effect of the lactobacilli on the natural background flora in the cottage cheese. For most experiments, as the numbers of lactobacilli increased, the numbers of spoilage organisms were lower than in the control on any given day of storage. In cheese inoculated with P. fluorescens, the numbers of spoilage organisms in the control had increased 5 log cycles by d 7, whereas the treatment containing the highest level of L. delbrueckii ssp. lactis RM2-5 (1.0 x 10(9) cfu/g) had not, and did not, increase over the course of the 21-d study. In the experiments where no spoilage organisms were added, lactobacilli significantly retarded the growth of gram-negative bacteria in the cheese. However, in these experiments, mold growth on the samples became a limiting factor during extended storage. The results of these experiments indicate that lactobacilli could be effective at helping control gram-negative spoilage bacteria in cottage cheese, thus potentially extending its shelf life. PMID- 15778301 TI - Processing effects on physicochemical properties of creams formulated with modified milk fat. AB - Type of thermal process [high temperature, short time pasteurization (HTST) or ultra-high temperature pasteurization (UHT)] and homogenization sequence (before or after pasteurization) were examined for influence on the physicochemical properties of natural cream (20% milk fat) and creams formulated with 20% low melt, fractionated butteroil emulsified with skim milk, or buttermilk and butter derived aqueous phase. Homogenization sequence influenced physicochemical makeup of the creams. Creams homogenized before pasteurization contained more milk fat surface material, higher phospholipid levels, and less protein at the milk fat interface than creams homogenized after pasteurization. Phosphodiesterase I activity was higher (relative to protein on lipid globule surface) when cream was homogenized before pasteurization. Creams formulated with skim milk and modified milk fat had relatively more phospholipid adsorbed at the milk fat interface. Ultra-high-temperature-pasteurized natural and reformulated creams were higher in viscosity at all shear rates investigated compared with HTST-pasteurized creams. High-temperature, short time-pasteurized natural cream was more viscous than HTST pasteurized reformulated creams at most shear rates investigated. High temperature, short time-pasteurized creams had better emulsion stability than UHT pasteurized creams. Cream formulated with buttermilk had creaming stability most comparable to natural cream, and cream formulated with skim milk and modified butteroil was least stable to creaming. Most creams feathered in a pH range of 5.00 to 5.20, indicating that they were moderately stable to slightly unstable emulsions. All processing sequences yielded creams within sensory specifications with the exception of treatments homogenized before UHT pasteurization and skim milk formulations homogenized after UHT pasteurization. PMID- 15778302 TI - Antioxidative activity of lactobacilli measured by oxygen radical absorbance capacity. AB - The reducing ability and antioxidative activity of some species of Lactobacillus were compared under in vitro conditions. Cultures of Lactobacillus delbrueckii ssp. lactis, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and Lactobacillus casei were grown at 37 degrees C in de Man, Rogosa, Sharpe (MRS) broth supplemented with 0.5% 2,3,5 triphenyl tetrazolium chloride (TTC) to evaluate reducing activity. Reduced TTC was extracted from the cultures with acetone, and the intensity of the red color measured colorimetrically at 485 nm was an indication of reducing activity. The lactobacilli varied significantly in relative ability to reduce TTC when grown in MRS broth for 15 h. The relative amounts of growth as indicated by pH values at 18 h appeared to influence the amount of reduction. Antioxidative activity was evaluated by the ability of the whole cells or the cell-free extracts from cultures to protect a protein from being attacked by free radicals. These analyses were performed using the oxygen radical absorbance capacity method. All cultures tested exhibited some degree of antioxidative activity. Among the treatments, the cell-free extracts from cells grown in MRS broth exhibited significantly higher values than did whole cells. There was no apparent relationship between the reducing and antioxidative activities of the cultures evaluated. The results from this study show that these cultures can provide a source of dietary antioxidants. Furthermore, selection of cultures that produce antioxidants as starters could provide yet another health or nutritional benefit from cultured or culture-containing dairy products. PMID- 15778303 TI - Impact of seasonal changes in ovine milk on composition and yield of a hard pressed cheese. AB - A hard-pressed, brined cheese was produced from frozen ovine milk collected in February, May, and August. Solids in the milk decreased as the season progressed. This was a result of high solids in early-lactation milk and low solids in August milk because of hot weather and poorer quality pastures. Casein as a percentage of true protein and the casein to fat ratio were higher in May and August milk. Fat in the cheese from February milk was higher and total protein was lower than in May and August. Milk, whey, and press whey composition were influenced by season and followed the trends of milk composition. Fat recovery in the cheeses ranged from 83.2 to 84.2%. Protein recovery in the cheeses was not affected by season. Cheese yield from February milk was higher than from May and August milk and was a result of higher casein and fat in the milk. PMID- 15778304 TI - Short communication: serum and tissue concentrations of vitamin D metabolites in beef heifers after buccal dosing of 25-hydroxyvitamin D3. AB - Sixteen crossbred (British x Continental; average un-shrunk body weight = 507.9 kg; SD = 45.6 kg) beef heifers fed a steam-flaked corn-based finishing diet with melengestrol acetate (0.4 mg/heifer daily) included to suppress estrus were used in a completely random design to evaluate the efficacy of buccal administration of 0, 10, 100, or 1000 mg of 25-hydroxyvitamin D3, (25-OH D3). Serum Ca, P, Mg, 25-OH D3, 1,25-dihydroxyvitamin D [1,25-(OH)2 D3], albumin, and protein were measured 24 h before dosing (-24 h), at dosing (0 h), and 6 and 24 h after dosing, after which the cattle were slaughtered at a commercial facility. Samples of kidneys, liver, longissimus lumborum, and triceps brachii were collected and evaluated for concentrations of 1,25-(OH)2 D3. With -24 and 0 h as baseline covariates, a significant time x treatment interaction was observed for serum 25 OH D3 and Ca concentrations, but not for serum 1,25-(OH)2 D3. Supplemental 25-OH D3 doses of 100 and 1000 mg significantly increased serum 25-OH D3 at 24 h after dosing, 1,25-(OH)2 D3 at 6 and 24 h after dosing, and serum Ca at 24 h after dosing. Similarly, buccal dosing of 1000 mg of supplemental 25-OH D3 significantly increased (approximately 2- to 3-fold) concentrations of 1,25-(OH)2 D3 in the kidney, liver, and longissimus lumborum relative to the other 3 treatments but not in triceps brachii. Serum albumin, protein, P, and Mg were not affected by treatment. Based on these results, buccal administration of 100 and 1000 mg 25-OH D3 increased vitamin D3 metabolites in serum and tissues, and it should be an effective method of delivering the vitamin. PMID- 15778305 TI - Expression of growth hormone receptor 1A mRNA is decreased in dairy cows but not in beef cows at parturition. AB - The promoter controlling expression of a major bovine growth hormone (GH) receptor (GHR) mRNA variant, GHR 1A, contains a common DNA element for transcription factors hepatocyte nuclear factor 4alpha (HNF-4alpha), hepatocyte nuclear factor 4gamma (HNF-4gamma), and chicken ovalbumin transcription factor II (COUP-TFII). Expression of GHR 1A mRNA is decreased in the liver of dairy cows at parturition. The objective of this study was to determine whether reduced expression of GHR 1A mRNA in dairy cows at parturition is associated with changed expression of HNF-4alpha, HNF-4gamma, or COUP-TFII mRNA. Liver biopsy samples were taken from multiparous Holstein cows 7 to 23 d before parturition, within 24 h after parturition (i.e., at parturition), and 8 to 18 d after parturition, and the relative amounts of GHR 1A, insulin-like growth factor-I (IGF-I), HNF-4alpha, HNF-4gamma, and COUP-TFII mRNA in these samples were measured by ribonuclease protection assays. As expected, expression of GHR 1A, total GHR, and IGF-I mRNA was decreased at parturition, compared with that detected prepartum or during the postpartum period. Expression of HNF-4alpha and COUP-TFII mRNA was unchanged, but that of HNF-4gamma mRNA was increased at parturition. The same study was also conducted in multiparous Angus cows 7 to 23 d before parturition, at parturition, and 8 to 18 d after parturition. Neither expression of GHR 1A, total GHR, or IGF I mRNA, nor expression of HNF-4alpha, COUP-TFII, or HNF-4gamma mRNA was changed in the liver of beef cows at parturition. These results together suggest that, at the molecular level, decreased expression of GHR 1A mRNA in the liver of dairy cows at parturition may involve increased expression of HNF-4gamma mRNA and that, at the systemic level, decreased expression of GHR 1A mRNA is not a direct result of the end of pregnancy, parturition, or the initiation of lactation. PMID- 15778306 TI - Molecular characterization of a saposin-like protein family member isolated from bovine lymphocytes. AB - Human and porcine T lymphocytes and natural killer (NK) cells produce antibacterial proteins that belong to the saposin-like family of proteins (SAPLIP). The objective of this study was to determine if a bovine homolog of SAPLIP exists in lymphocytes that exhibit antibacterial activity. Following stimulation with IL-2, bactericidal activity against Staphylococcus aureus was detected to some extent in most major subpopulations of T lymphocytes including CD4+, CD8+, CD3+, and WC1+ gammadelta T lymphocytes. However, the majority of antibacterial activity was observed in the CD2+CD3- lymphocytes, which are similar phenotypically to NK cells. A partial sequence of a bovine SAPLIP was generated using low specificity primers designed from regions of homology between other SAPLIP including porcine NK-lysin and human granulysin. Enhanced expression of the bovine lysin gene was detected in mRNA isolated from IL-2-stimulated CD2+CD3- lymphocytes. The partial cDNA sequence was then used to make gene specific primers for a rapid amplification of cDNA ends (RACE) procedure that provided repeatable 5' and 3' cDNA ends. By examining overlapping regions from the RACE procedure, full-length sequence information was obtained for the bovine lysin homologue. Conceptual translation of the cDNA demonstrated conserved similarities to known SAPLIP members. Further characterization of the bovine lysin may facilitate its use in protecting dairy cattle against bacterial infections. PMID- 15778307 TI - Presence of multidrug-resistant enteric bacteria in dairy farm topsoil. AB - In addition to human and veterinary medicine, antibiotics are extensively used in agricultural settings, such as for treatment of infections, growth enhancement, and prophylaxis in food animals, leading to selection of drug and multidrug resistant bacteria. To help circumvent the problem of bacterial antibiotic resistance, it is first necessary to understand the scope of the problem. However, it is not fully understood how widespread antibiotic-resistant bacteria are in agricultural settings. The lack of such surveillance data is especially evident in dairy farm environments, such as soil. It is also unknown to what extent various physiological modulators, such as salicylate, a component of aspirin and known model modulator of multiple antibiotic resistance (mar) genes, influence bacterial multi-drug resistance. We isolated and identified enteric soil bacteria from local dairy farms within Roosevelt County, NM, determined the resistance profiles to antibiotics associated with mar, such as chloramphenicol, nalidixic acid, penicillin G, and tetracycline. We then purified and characterized plasmid DNA and detected mar phenotypic activity. The minimal inhibitory concentrations (MIC) of antibiotics for the isolates ranged from 6 to >50 microg/mL for chloramphenicol, 2 to 8 microg/mL for nalidixic acid, 25 to >300 microg/mL for penicillin G, and 1 to >80 microg/mL for tetracycline. On the other hand, many of the isolates had significantly enhanced MIC for the same antibiotics in the presence of 5 mM salicylate. Plasmid DNA extracted from 12 randomly chosen isolates ranged in size from 6 to 12.5 kb and, in several cases, conferred resistance to chloramphenicol and penicillin G. It is concluded that enteric bacteria from dairy farm topsoil are multidrug resistant and harbor antibiotic-resistance plasmids. A role for dairy topsoil in zoonoses is suggested, implicating this environment as a reservoir for development of bacterial resistance against clinically relevant antibiotics. PMID- 15778308 TI - Subacute ruminal acidosis induces ruminal lipopolysaccharide endotoxin release and triggers an inflammatory response. AB - Subacute ruminal acidosis (SARA) was induced in 3 rumen fistulated Jersey steers by offering them different combinations of wheat-barley pellets and chopped alfalfa hay. Steers were offered 4, 5, and 6 kg/d of pelleted concentrate and 6, 5, and 4 kg/d of chopped alfalfa hay for diets 1, 2, and 3, respectively, during 5-d treatment periods and were fed chopped alfalfa hay between treatment periods. Inducing SARA increased blood concentrations of haptoglobin and serum amyloid-A. Dry matter intake of concentrate and hay decreased from d 1 to 5 in each period. Subacute ruminal acidosis was induced in all steers during d 4 and 5 when concentrate was fed, with ruminal pH remaining below 5.6 for an average of 187 and 174 min/d on these days. Lipopolysaccharide concentration increased significantly during periods of grain feeding compared with times when only hay was fed. Inducing SARA by feeding wheat-barley pellets activated a systemic inflammatory response in the steers. PMID- 15778309 TI - Effects of yeast culture and galacto-oligosaccharides on ruminal fermentation in holstein cows. AB - Four nonlactating, ruminally cannulated Holstein cows were used in a 4 x 4 Latin square design, balanced for residual effects, to evaluate the effects of supplementing dairy cow diets with yeast culture (Trichosporon sericeum; YC), galacto-oligosaccharides (GOS), or the mixture of YC and GOS on ruminal fermentation, microbial N supply, in situ degradation, and energy and nitrogen metabolism. Treatments were arranged in a 2 x 2 factorial as follows: 1) basal diet, 2) basal diet plus 10 g/d YC, 3) basal diet plus 2% GOS, 4) basal diet plus a mixture of 10 g/d YC and 2% GOS. Nitrogen losses in urine were lower, and retained N was higher, for cows supplemented with a mixture of YC and GOS. Ruminal pH was lower in cows supplemented with GOS alone compared with other treatments. Total VFA concentration was higher in cows fed control and GOS supplemented diets than in those fed YC containing diets. The molar proportion of propionate was higher, and the molar proportion of acetate was lower, in cows fed control diets. Microbial N supply was higher in cows fed control diets. There were no major positive effects of supplements observed in this study. However, supplementation of a mixture of YC and GOS had a tendency for synergistic effects on N metabolism and in situ degradation of a soluble fraction of oat straw DM and CP of concentrates compared with supplementation of YC or GOS alone. PMID- 15778310 TI - Corn grain endosperm type and brown midrib 3 corn silage: site of digestion and ruminal digestion kinetics in lactating cows. AB - Interactions of endosperm type of corn grain and the brown midrib 3 (bm3) mutation in corn silage on ruminal kinetics and site of nutrient digestion of lactating dairy cows were evaluated. Eight ruminally and duodenally cannulated cows (72 +/- 8 d in milk; mean +/- SD) were used in a duplicated 4 x 4 Latin square design experiment with a 2 x 2 factorial arrangement of treatments. Treatments were corn grain endosperm type (floury or vitreous) and corn silage type (bm3 or isogenic normal). Diets contained 26% neutral detergent fiber (NDF) and 30% starch. Interactions of treatments were not observed for any measure of digestibility, but digestion kinetics of starch and fiber did interact to affect digestible organic matter intake by affecting dry matter intake. Rate of ruminal starch digestion was faster and rate of ruminal starch passage tended to be slower in diets containing corn grain with floury vs. vitreous endosperm, resulting in a mean increase of 22 units for ruminal starch digestibility. Although compensatory postruminal starch digestion decreased differences among treatments for total tract starch digestibility, starch entering the duodenum was more digestible for grain with floury endosperm compared with vitreous grain, resulting in greater total tract starch digestibility for floury compared with vitreous corn grain. Fermentation rate of potentially digestible NDF was not affected by either bm3 corn silage or greater ruminal starch digestion of floury grain. Brown midrib corn silage increased total tract NDF digestibility vs. control silage by numerically increasing ruminal and postruminal digestibility of NDF. Endosperm type of corn grain greatly influences site of starch digestion and should be considered when formulating diets. PMID- 15778311 TI - Corn grain endosperm type and brown midrib 3 corn silage: feeding behavior and milk yield of lactating cows. AB - Interactions of endosperm type of corn grain and the brown midrib 3 mutation (bm3) in corn silage on feeding behavior, productivity, energy balance, and plasma metabolites of lactating dairy cows were evaluated. Eight ruminally and duodenally cannulated cows (72 +/- 8 d in milk; mean +/- SD) were used in a duplicated 4 x 4 Latin square design experiment with a 2 x 2 factorial arrangement of treatments. Treatments were corn grain endosperm type (floury or vitreous), and corn silage type (bm3 or isogenic control). Diets contained 26% neutral detergent fiber (NDF) and 30% starch. Floury endosperm grain decreased dry matter intake (DMI) 1.9 kg/ d compared with vitreous grain when combined with control corn silage but did not affect DMI when combined with bm3 corn silage. This interaction of treatments occurred because of changes in meal size; floury endosperm grain decreased meal size in control silage diets but increased meal size in bm3 corn silage diets. Ruminal pool sizes reflected DMI differences among diets, suggesting that ruminal fill was not the primary limitation on intake. Brown midrib 3 corn silage reduced rumination time per day and number of rumination bouts per day. Floury endosperm grain decreased 3.5% fat-corrected milk by 1.2 kg/d when combined with control silage but increased 3.5% fat corrected milk by 2.1 kg/d when combined with bm3 corn silage. Starch and fiber digestibility interact to affect feeding behavior and milk production and production response to bm3 corn silage depends on the grain source that is fed. PMID- 15778312 TI - Corn grain endosperm type and brown midrib 3 corn silage: ruminal fermentation and N partitioning in lactating cows. AB - Interactions of endosperm type of corn grain and the brown midrib 3 mutation (bm3) in corn silage on ruminal fermentation and microbial efficiency of lactating dairy cows were evaluated. Eight ruminally and duodenally cannulated cows (72 +/- 8 d in milk; mean +/- SD) were used in a duplicated 4 x 4 Latin square design experiment with a 2 x 2 factorial arrangement of treatments. Treatments were corn grain endosperm type (floury or vitreous) and corn silage type (bm3 or isogenic normal). Diets contained 26% neutral detergent fiber and 30% starch. Increasing ruminal starch digestibility by replacing vitreous corn grain with floury grain reduced mean and minimum ruminal pH. Brown midrib 3 corn silage reduced mean and minimum ruminal pH and increased total volatile fatty acid concentration. Ruminal pH was positively associated with rate of valerate absorption. Although floury endosperm reduced acetate:propionate ratio in both control and bm3 corn silage diets, it had a greater effect on reducing acetate:propionate ratio for control silage compared with bm3 corn silage. Nonammonia N flow to the duodenum did not differ among treatments and no effects of treatment were detected for microbial N and nonammonia, nonmicrobial N flow. Although treatment effects on ruminal fermentation and ruminal pH were observed, few interactions of treatment were detected and treatments did not affect flow of N fractions to the intestines. PMID- 15778313 TI - The effect of concentrate supplementation on nutrient flow to the omasum in dairy cows receiving freshly cut grass. AB - An experiment was carried out to determine the effect of increasing the amount of grain-based concentrate (0, 3, or 6 kg/d) on nutrient flow to the omasum, rumen fermentation pattern, milk yield, and nutrient use of dairy cows. Harvested timothy-meadow fescue grass was fed individually 3 times daily to 6 rumen cannulated Holstein-Friesian cows in a duplicated 3 x 3 Latin square experiment. Grass was offered as 6 equal meals daily, and concentrates were fed as 2 equal meals daily. Nitrogen, microbial N, and neutral detergent fiber (NDF) flow from the rumen were measured using an omasal sampling technique in combination with a triple marker method [CoEDTA, Yb, and indigestible NDF (INDF) as markers]. Concentrate supplementation linearly decreased ruminal pH, N degradability, ammonia N concentration, and molar proportion of acetate and increased the molar proportion of butyrate. Supplementation of grass with concentrates linearly increased dry matter intake (DMI), microbial N synthesis, N, and NDF flow to the omasum, and ruminal and total tract NDF digestibility decreased linearly. Decreases in NDF digestibility in response to concentrates was primarily related to a decrease in the rate of digestion. Increased DMI overcame the negative effects of concentrate on NDF digestion, resulting in a linear increase in total metabolizable energy intake and milk production. Physical constraints were found not to limit grass DMI. Concentrate supplementation increased the apparent use of dietary N for milk production because of a reduction in N intake, rather than thorough improvements in N capture in the rumen. PMID- 15778314 TI - Calf response to caustic paste and hot-iron dehorning using sedation with and without local anesthetic. AB - Dairy producers require effective methods of reducing the pain associated with dehorning. Previous work has shown that analgesics can reduce pain associated with hot-iron dehorning, but these interventions may not be practical for some producers. Some producers favor the use of caustic paste (to cause a chemical burn as opposed to thermal burn), but little is known about how to treat pain caused by caustic burns. In 2 experiments, head shaking and head rubbing behaviors were used to evaluate pain following dehorning using caustic paste. In experiment 1, sedated calves were dehorned using a caustic paste, with or without a lidocaine local block. Calves treated with lidocaine showed no evidence of reduced pain response in the hours after the caustic paste was applied. In experiment 2, response to caustic paste dehorning with a sedative only was compared with hot-iron dehorning using a sedative and local anesthetic. In the first 4 h after dehorning, calves dehorned with a hot iron showed more head shaking than calves dehorned with caustic paste. These results indicate that dehorning with caustic paste combined with a sedative results in less pain to calves than dehorning with a hot iron combined with a sedative and a local anesthetic. PMID- 15778315 TI - Effect of nursing management and skeletal size at weaning on puberty, skeletal growth rate, and milk production during first lactation of dairy heifers. AB - Forty Israeli-Holstein 5-d-old calves were used to determine the effect of increasing calf body weight (BW) and skeletal size during the nursing period on age and skeletal size at puberty and on skeletal size and performance during first lactation. The calves were randomly allotted to 2 experimental groups as follows: milk replacer (MR) [calves were given 0.450 kg/d dry matter of milk replacer for the first 50 d of life] and milk-fed (MF) [calves had free access to milk in two 30-min meals/d]. From weaning to 180 d of age, all calves were fed the same diet. At 180 d of age, the MR and MF calves were each divided into 2 equal subgroups: one subgroup from each treatment was given only growing ration, and the other was given the same ration supplemented with fish meal to supply 2% crude protein (CP) (treatments MR + CP and MF + CP, respectively). Finally, at 270 d of age, all calves were housed together and fed a growing heifer's ration until first calving. During the entire nursing period, the MF calves consumed 9.8% more DM, 39.7% more CP, and 52.4% more metabolizable energy than the MR calves. At 60 d of age, BW and all skeletal parameters were higher in the MF calves than in the MR calves. During the entire rearing period (60 to 550 d), the average BW of the MF calves was greater by 16 kg than the BW of the MR calves. Nursing management did not affect differences in skeletal parameters at calving. Average age at puberty onset was 272 +/- 26.8 d; MF calves reached puberty 23 d earlier than MR calves. Yields of milk (kg/305 d) and fat-corrected milk (FCM, kg/d) were greater for the MF + CP heifers than for the MR heifers. It was concluded that nursing by ad libitum milk, as compared with milk replacer, affected BW but not skeletal size of the adult animal, decreased age of puberty onset, and increased FCM yield at first lactation. Supplementing the diet with 2% CP during the prepubertal period increased BW but not skeletal size of the adult animal and 305-d milk and FCM yields during first lactation. PMID- 15778316 TI - Nutrient content of whole cottonseed. AB - The objective of this study was to determine if the nutrient and gossypol contents and in vitro digestibility of 3 types of genetically modified whole cottonseed differed from traditional whole cottonseed. Samples of seed from traditional (no genetic modifications) and genetically modified varieties of cotton grown in 1999 and 2000 were analyzed. Genetic modifications included the insertion of genes to protect cotton from insect pests (Bt), and damage from glyphosate herbicides (RR), and from both (Bt/RR). Year effects were significant for in vitro dry matter (DM) digestibility, gossypol, DM, crude protein (CP), fat, neutral detergent fiber (NDF), acid detergent fiber (ADF), and ash. Higher rainfall resulted in higher CP, fat, and ash and lower NDF and gossypol. There were no differences among seed types for ground or whole seed digestibility, DM, CP, fat, NDF, ADF, ash, lignin, net energy for lactation, amino acids, total fatty acids, or seed index. Overall, the nutrient content and digestibility of varieties of genetically modified seed were similar to that of varieties of traditional whole cottonseed. PMID- 15778317 TI - Mammary lipid metabolism and milk fatty acid secretion in alpine goats fed vegetable lipids. AB - Fourteen Alpine goats at midlactation were fed a diet of hay and concentrate (55:45), without (control) or with formaldehyde-treated linseed (FLS) or oleic sunflower oil (OSO) at 11.2 or 3.5% of dry matter intake, respectively, in a 3 x 3 Latin Square design with three 3-wk periods. Milk yield was lower in goats fed FLS than control or OSO (2.13 vs. 2.32 kg/d). Milk fat content was higher with FLS or OSO than control (40.8 vs. 33.8 g/kg). Formaldehyde-treated linseed and OSO caused a significant decrease (23 and 18%, respectively) of C10 to C17 fatty acids secretion compared with control. The secretion of cis-9 C18:1 and cis-9, trans-11 C18:2 were increased 1.44- and 1.54-fold for FLS and 1.78- and 1.36-fold for OSO, compared with control. The C18:3 (n-3) secretion was increased 2.61-fold with FLS compared with control. Milk cis-9 C14:1/C14:0, cis-9 C16:1/C16:0, and cis-9 C18:1/C18:0 ratios decreased with the supplemented diets compared with control. Mammary stearoyl-CoA desaturase mRNA and activity were decreased by the lipid supplements, whereas no significant change was observed for acetyl-CoA carboxylase and fatty acid synthase. The activities of glucose-6-phosphate dehydrogenase, malic enzyme, and glycerol-3-phosphate dehydrogenase were not affected by the lipid supplements. Mammary lipoprotein lipase mRNA increased with OSO, whereas lipoprotein lipase activity tended to decrease with FLS compared with control. Milk lipoprotein lipase activity sharply decreased with lipid supplement (by 59 and 71%, for FLS and OSO, respectively). The changes in milk fatty acid profile due to FLS and OSO supplements were partly related to changes in the levels of mammary enzyme activities or mRNA. PMID- 15778318 TI - Caprine kappa-casein (CSN3) polymorphism: new developments in molecular knowledge. AB - A high degree of polymorphism was recently found at the kappa-casein (CSN3) locus in the domesticated goat (Capra hircus). In the present study, 2 new patterns previously identified by PCR-single-strand conformation polymorphism analysis (SSCP) were characterized. The allele provisionally named "X" (GenBank Accession no. AY350425) differs from CSN3*C (AF485341) by a (silent) A-->G substitution at position 509 of the goat CSN3 reference sequence (X60763). As this newly identified sequence changes the amino acid sequence, and the already known CSN3*C allele (AF485341) has an additional silent mutation, we proposed a change in nomenclature to reflect these changes, indicating the silent mutation with the prime symbol (i.e.,'). The CSN3*M allele (provisionally named "Y") results in a new protein variant, differing by 2 nonsynonymous mutations from the CSN3*F allele. The new variant is characterized by a G-->A transition at nucleotide position 384, resulting in the amino acid exchange Asp90-->Asn90, and a C-->T transition at position 550, resulting in a Val145-->Ala145 substitution. Thus, the number of alleles identified in the domesticated goat has increased to 16, of which 13 are protein variants and 3 are silent mutations, involving a total of 15 polymorphic sites in CSN3 exon 4. Data on the distribution of the main alleles in 7 goat breeds of Europe, West Africa, and the Near East show differences in the occurrence and frequency of the alleles between breeds and geographic origin with the highest number of alleles found in goat breeds from the Near East. PMID- 15778319 TI - Methodology for estimation of days dry effects. AB - The primary objective of this research was to determine if, with appropriate methodology, unbiased estimates of days dry (DD) effects on subsequent lactation milk yield can be obtained from field data, particularly when DD is correlated with cow effects. Another objective was to ascertain relevant sampling properties of designed trials for estimation of DD effects. Simulated records were used to assess methodology. Along with a model with no adjustments for cow effects, alternative models including 1) previous lactation milk yield, 2) a prior adjustment for cow effects estimated from an animal model, and 3) a combination of 1 and 2, were tested. Estimates from the unadjusted model were biased downward; however, the 3 alternative analyses provided estimates of DD effects that were essentially unbiased, with a prior adjustment for cow effects and previous milk yield in the model providing the best results in terms of elimination of bias. Therefore, DD effects can be estimated from field data without bias from cow effects.A designed trial with 2 groups and 10 or fewer cows/ group is noninformative and has an unacceptably high probability of leading to invalid conclusions. A minimum of 30 cows/group is considerably better and should be used whenever possible. Even with 30 cows/group, however, the power is low unless the difference between DD groups for yield is at least 1130 kg. Prior correction of 305-d, mature equivalent records for cow effects, using predicted producing abilities, could be done in designed trials to improve the statistical power of tests and accuracy of estimates. PMID- 15778320 TI - Genetic association between susceptibility to clinical mastitis and protein yield in norwegian dairy cattle. AB - The objective of this study was to examine associations between susceptibility to clinical mastitis and protein yield in first-lactation Norwegian Dairy Cattle (NRF) cows. Records from 372,227 first-lactation daughters of 2411 NRF sires were analyzed bivariately, using a threshold-liability model for clinical mastitis and a linear Gaussian model for 305-d protein yield. The mean (SD) of the posterior distribution of heritability was 0.08 (0.004) for susceptibility to clinical mastitis and 0.19 (0.007) for 305-d protein yield. The posterior mean (SD) of the genetic correlation between susceptibility to clinical mastitis and 305-d protein yield was 0.43 (0.03). Posterior means of the correlations between herd-5-yr effects, and between model residuals were 0.19 and -0.008, respectively. Corresponding estimates of genetic, herd-5-yr, and residual correlations from a bivariate linear model analysis were 0.42, 0.18, and -0.008, respectively. An antagonistic genetic relationship between clinical mastitis and protein yield was corroborated. PMID- 15778321 TI - Genetic improvement in mastitis resistance: comparison of selection criteria from cross-sectional and random regression sire models for somatic cell score. AB - Several selection criteria for reducing incidence of mastitis were developed from a random regression sire model for test-day somatic cell score (SCS). For comparison, sire transmitting abilities were also predicted based on a cross sectional model for lactation mean SCS. Only first-crop daughters were used in genetic evaluation of SCS, and the different selection criteria were compared based on their correlation with incidence of clinical mastitis in second-crop daughters (measured as mean daughter deviations). Selection criteria were predicted based on both complete and reduced first-crop daughter groups (261 or 65 daughters per sire, respectively). For complete daughter groups, predicted transmitting abilities at around 30 d in milk showed the best predictive ability for incidence of clinical mastitis, closely followed by average predicted transmitting abilities over the entire lactation. Both of these criteria were derived from the random regression model. These selection criteria improved accuracy of selection by approximately 2% relative to a cross-sectional model. However, for reduced daughter groups, the cross-sectional model yielded increased predictive ability compared with the selection criteria based on the random regression model. This result may be explained by the cross-sectional model being more robust, i.e., less sensitive to precision of (co)variance components estimates and effects of data structure. PMID- 15778322 TI - The relationship between fertility, rump angle, and selected type information in Holstein-Friesian cows. AB - Some dairy cattle breeders believe that dairy cows with high pin bones will have poorer fertility. The aim of this study was to examine this claim by investigating the relationships, both genetic and phenotypic, between fertility, rump angle, and other selected type traits in first-lactation Holstein-Friesians. Results showed an unfavorable genetic correlation (-0.16) between calving interval and rump angle, suggesting that animals with high pin bones would have a longer calving interval. However, no significant genetic or phenotypic correlation between days to first service and nonreturn rate and rump angle was observed. No evidence of a relationship, linear or quadratic, between any fertility trait and rump angle was found. Udder support and mammary system were unfavorably correlated to calving interval (0.25 and 0.14, respectively), suggesting that cows with more functional udders would have a longer calving interval. Legs and feet score was favorably correlated to nonreturn rate, suggesting that animals with good legs and feet would be less likely to return to service. PMID- 15778323 TI - Genetic evaluation of calving ease for Brown Swiss and Jersey bulls from purebred and crossbred calvings. AB - The objective of this study was to examine the feasibility of implementing routine national calving ease (CE) genetic evaluations of Brown Swiss (BS) and Jersey (JE) sires that include records of crossbred calvings. Records were available for 11,793 BS calvings, 3431 BS-sired crosses, 65,293 JE calvings, and 7090 JE-sired crosses. Evaluations were performed for each breed using only purebred calvings and using both purebred and crossbred calvings. In the latter evaluations, the sire-maternal grandsire model used for the routine evaluation of Holstein (HO) CE was modified to include a fixed breed composition effect to account for differences between purebred and crossbred calvings. Jersey cows had very little calving difficulty (0.5 to 0.7%) and JE bulls had a very small range of evaluations, suggesting that a routine JE evaluation would be of little value. Results from the BS evaluations suggest a routine evaluation would provide BS breeders with a useful tool for genetic improvement. Further examination of data showed that many BS calvings were in mixed herds with HO calvings. As a result, a joint evaluation for BS and HO bulls was developed. The BS data showed that there is similar genetic variability as found in the HO population, which suggests implementation of a routine evaluation including BS CE would be of value. It appears BS bulls may produce daughters with superior maternal calving ability compared with HO. Validation of the joint evaluation was performed by comparing results with the routine HO evaluation. Holstein solutions from the joint evaluation were comparable to results from the routine HO-only evaluation. Correlations among solutions and evaluations showed HO evaluations were not adversely affected by BS data and BS sires were reranked as compared with the BS only evaluation. PMID- 15778324 TI - Genetics of parity-dependant production increase and its relationship with health, fertility, longevity, and conformation in swiss holsteins. AB - Genetic analysis of production increase (ProdI), defined as an increase in production from early to later lactations, was conducted using data from the Holstein Association of Switzerland. This production increase describes the maturity rate of the cow. The data set contained 42,807 cows with a ProdI value. All cows had completed the first 3 lactations. Different formulas were derived for the computation of ProdI using 1) milk yields or energy-corrected milk yields and 2) yields from all 3 lactations or only 2 of them (first and second, first and third, second and third). Heritabilities of ProdI and genetic and phenotypic correlations of ProdI with somatic cell score, days to first service, nonreturn rate, longevity, and 27 conformation traits were estimated by univariate and bivariate sire models that included relationship among sires. Heritabilities for ProdI were low (0.06 to 0.08), but genetic variation among sires existed. For nonreturn rate and longevity, regressions on the sire estimated breeding values were estimated. Additive genetic correlations of ProdI were moderately favorable with somatic cell score (-0.22 to -0.33) and chest width (0.21 to 0.30), i.e., with traits often associated with long-lasting cows. Unfavorable correlations were found with angularity (-0.18 to -0.26). Regression coefficients from regressing ProdI on sire estimated breeding values for longevity tend to show favorable relationships between these 2 traits (0.10 to 0.20). Results show that animals can be selected for ProdI, as there is good genetic variation between bulls. ProdI is a potential trait to be included in selection indices, as it has favorable genetic relationships with economically important functional traits such as health, conformation, and longevity. PMID- 15778325 TI - Relationship between type traits and longevity in Canadian Jerseys and Ayrshires using a Weibull proportional hazards model. AB - The aim of this study was to use a Weibull proportional hazards model to explore the impact of type traits on the functional survival of Canadian Jersey and Ayrshire cows. The data set consisted of 49,791 registered Jersey cows from 900 herds calving from 1985 to 2003. The corresponding figures for Ayrshire were 77,109 cows and 921 herds. Functional survival was defined as the number of days from first calving to culling, death, or censoring. Type information consisted of phenotypic type scores for 8 composite traits and 19 linear descriptive traits. The statistical model included the effects of stage of lactation; season of production; annual change in herd size; type of milk recording supervision; age at first calving; effects of milk, fat, and protein yields calculated as within herd-year-parity deviations; herd-year-season of calving; each type trait; and the animal's sire. Analysis was done one trait at a time for each of 27 type traits in each breed. The relative culling risk was calculated for animals in each class after accounting for the previously mentioned effects. Among the composite type traits with the greatest contribution to the likelihood function was final score followed by mammary system for Jersey breed, while in Ayrshire breed feet and legs was the second most important trait next to final score. Cows classified as Poor for final score in both breeds were >5 times more likely to be culled compared with the cows classified as Good Plus. In both breeds, cows classified as Poor for feet and legs were 5 times more likely to be culled than were cows classified as Excellent, and cows classified as Excellent for mammary system were >9 times more likely to survive than were cows classified as Poor. PMID- 15778326 TI - Casein haplotype structure in five Italian goat breeds. AB - The aim of this work was to investigate the genetic structure of the casein gene cluster in 5 Italian goat breeds and to evaluate the haplotype variability within and among populations. A total of 430 goats from Vallesana, Roccaverano, Jonica, Garganica, and Maltese breeds were genotyped at alphas1-casein (CSN1S1), alphas2 casein, (CSN1S2), beta-casein (CSN2), and kappa-casein (CSN3) loci using several genomic techniques and milk protein analysis. Casein haplotype frequencies were estimated for each breed. Principal component analysis was carried out to highlight the relationship among breeds. Allele and haplotype distributions indicated considerable differences among breeds. The haplotype CSN1S1*F- CSN1S2*F CSN3*D occurred in all breeds with frequencies >0.100 and was the most common haplotype in the Southern breeds. A high frequency of CSN1S1*0-CSN1S2*C-CSN3*A haplotype was found in Vallesana population (0.162). Principal component analysis clearly separated the Northern and Southern breeds by the first component. The variability of the caprine casein loci and variety of resulting haplotypes should be exploited in the future using specific breeding programs aiming to preserve biodiversity and to select goat genetic lines for specific protein production. PMID- 15778327 TI - Genetic progress in multistage dairy cattle breeding schemes using genetic markers. AB - The aim of this paper was to explore general characteristics of multistage breeding schemes and to evaluate multistage dairy cattle breeding schemes that use information on quantitative trait loci (QTL). Evaluation was either for additional genetic response or for reduction in number of progeny-tested bulls while maintaining the same response. The reduction in response in multistage breeding schemes relative to comparable single-stage breeding schemes (i.e., with the same overall selection intensity and the same amount of information in the final stage of selection) depended on the overall selection intensity, the selection intensity in the various stages of the breeding scheme, and the ratio of the accuracies of selection in the various stages of the breeding scheme. When overall selection intensity was constant, reduction in response increased with increasing selection intensity in the first stage. The decrease in response was highest in schemes with lower overall selection intensity. Reduction in response was limited in schemes with low to average emphasis on first-stage selection, especially if the accuracy of selection in the first stage was relatively high compared with the accuracy in the final stage. Closed nucleus breeding schemes in dairy cattle that use information on QTL were evaluated by deterministic simulation. In the base scheme, the selection index consisted of pedigree information and own performance (dams), or pedigree information and performance of 100 daughters (sires). In alternative breeding schemes, information on a QTL was accounted for by simulating an additional index trait. The fraction of the variance explained by the QTL determined the correlation between the additional index trait and the breeding goal trait. Response in progeny test schemes relative to a base breeding scheme without QTL information ranged from +4.5% (QTL explaining 5% of the additive genetic variance) to +21.2% (QTL explaining 50% of the additive genetic variance). A QTL explaining 5% of the additive genetic variance allowed a 35% reduction in the number of progeny tested bulls, while maintaining genetic response at the level of the base scheme. Genetic progress was up to 31.3% higher for schemes with increased embryo production and selection of embryos based on QTL information. The challenge for breeding organizations is to find the optimum breeding program with regard to additional genetic progress and additional (or reduced) cost. PMID- 15778328 TI - Evaluation of closed adult nucleus multiple ovulation and embryo transfer and conventional progeny testing breeding schemes for milk production in tropical crossbred cattle. AB - The potential benefits of closed adult nucleus multiple ovulation and embryo transfer (MOET) and conventional progeny testing (CNS) schemes, and the logistics of their integration into large-scale continuous production of crossbred cattle were studied by deterministic simulation. The latter was based on F1 (Bos taurus x Bos indicus) production using AI or natural mating and MOET, and continuous F2 production by mating of F1 animals. The gene flow and the cumulative discounted expressions (CDES) were also calculated. Both schemes had 8, 16, 32, or 64 dams with 2, 4, 8, 16, or 32 sires selected. In the MOET nucleus scheme (MNS), the test capacity was 1, 2, 8, or 16 offspring, and the number of matings per dam per year was 1, 2, or 4. A scheme of 8 sires with 64 dams and a test capacity of 4 female offspring per dam per year resulted in an annual genetic gain (in phenotypic standard deviation) of 0.324 and 0.081 for MNS and CNS, respectively. In the MNS, there was substantial genetic gain with a relatively small number of animals compared with a CNS. The F1 had the highest, and the F2 scheme the lowest CDES. However, a very large number of B. indicus females would be required in the F1 scheme. This scheme may not be practical under conditions in developing countries. The F2 scheme was logistically attractive because it produces its own replacements, and the number of B. taurus females required would be easy to attain. Accompanying technical and financial constraints of nucleus schemes should be addressed before applying them. PMID- 15778329 TI - A comparison of three animal welfare assessment programs on California dairies. AB - Due to increased awareness of animal welfare issues by consumers, 3 voluntary programs were developed to assess animal welfare on commercial dairies in the United States. These programs include: 1) Humane Farm Animal Care (HFAC), with minimum standards and a third-party audit system; 2) Dairy Quality Assurance Center (DQAC); and 3) University of California-Davis (UCD) program with self assessments. The objective of this article was to provide an overview of the similarities and differences in content and scope of these programs. Furthermore, to ascertain the practical application of these programs on commercial dairies, a study was designed to compare the rank order of compliance scores of each program on 10 commercial California dairies using a survey and personal interviews with dairy owners or managers. The survey combined 316 statements from the DQAC checklist, the UCD assessment, and the HFAC standards. The mean (+/-SD) percentage compliance for the DQAC, HFAC, and UCD programs was 83 +/- 8, 94 +/- 3, and 85 +/- 6%, respectively. A ranking order from 1 to 10 was assigned to each dairy using the compliance score for each assessment program. Comparisons of ranking order between programs were not significant, indicating that different indices were being measured by each program. All programs, however, identified the same 2 lowest-ranking dairies, which indicated that the programs were able to identify facilities with the weakest welfare practices. The survey results show that selection of the available assessment programs for animal welfare on the dairy is important in determining the outcomes. PMID- 15778330 TI - Improving food safety within the dairy chain: an application of conjoint analysis. AB - This study determined the relative importance of attributes of food safety improvement in the production chain of fluid pasteurized milk. The chain was divided into 4 blocks: "feed" (compound feed production and its transport), "farm" (dairy farm), "dairy processing" (transport and processing of raw milk, delivery of pasteurized milk), and "consumer" (retailer/catering establishment and pasteurized milk consumption). The concept of food safety improvement focused on 2 main groups of hazards: chemical (antibiotics and dioxin) and microbiological (Salmonella, Escherichia coli, Mycobacterium paratuberculosis, and Staphylococcus aureus). Adaptive conjoint analysis was used to investigate food safety experts' perceptions of the attributes' importance. Preference data from individual experts (n = 24) on 101 attributes along the chain were collected in a computer-interactive mode. Experts perceived the attributes from the "feed" and "farm" blocks as being more vital for controlling the chemical hazards; whereas the attributes from the "farm" and "dairy processing" were considered more vital for controlling the microbiological hazards. For the chemical hazards, "identification of treated cows" and "quality assurance system of compound feed manufacturers" were considered the most important attributes. For the microbiological hazards, these were "manure supply source" and "action in salmonellosis and M. paratuberculosis cases". The rather high importance of attributes relating to quality assurance and traceability systems of the chain participants indicates that participants look for food safety assurance from the preceding participants. This information has substantial decision-making implications for private businesses along the chain and for the government regarding the food safety improvement of fluid pasteurized milk. PMID- 15778332 TI - A multi-species epidemic model with spatial dynamics. AB - A model is formulated that describes the spatial propagation of a disease that can be transmitted between multiple species. The spatial component consists, for each species, of a certain number of patches that make up the vertices of a digraph, the arcs of which represent the movement of the various species between the patches. In each of the patches and for each species, a susceptible-exposed infectious-recovered (SEIR) epidemic model describes the evolution of the disease status of individuals. Also in each patch, there is transmission of the disease from species to species. An analysis of the system is given, beginning with results on the mobility component. A formula is derived for the computation of the basic reproduction number R(0) for sspecies and npatches, which then determines the global stability properties of the disease free equilibrium. Simulations for the spread of a disease in one species and two patches are presented. PMID- 15778333 TI - Simple models of antibiotic cycling. AB - The use of environmental heterogeneity is an old but potentially powerful method for managing biological systems. Determining the optimal form of environmental heterogeneity is a difficult problem. One family of heterogeneous management strategies that has received attention in the medical community is the periodic cycling of antibiotic usage to control antibiotic resistance. This paper presents a theory for the optimization of antibiotic cycling based on a density independent model of transmission and immigration of evolutionarily static strains. In the case of two pathogen strains, I show that the population's asymptotic growth rate is a monotonically increasing function of the oscillation period under certain common assumptions. Monte Carlo simulations show that this result fails in more general settings, but suggest that antibiotic cycling seldom provides a significant improvement over alternative mixing practices. The results support the findings of other researchers that antibiotic cycling does not offer significant advantages over idealized conventional practice. However, cycling strategies may be preferable in some special cases. PMID- 15778334 TI - Non-linear incidence and stability of infectious disease models. AB - In this paper we consider the impact of the form of the non-linearity of the infectious disease incidence rate on the dynamics of epidemiological models. We consider a very general form of the non-linear incidence rate (in fact, we assumed that the incidence rate is given by an arbitrary function f (S, I, N) constrained by a few biologically feasible conditions) and a variety of epidemiological models. We show that under the constant population size assumption, these models exhibit asymptotically stable steady states. Precisely, we demonstrate that the concavity of the incidence rate with respect to the number of infective individuals is a sufficient condition for stability. If the incidence rate is concave in the number of the infectives, the models we consider have either a unique and stable endemic equilibrium state or no endemic equilibrium state at all; in the latter case the infection-free equilibrium state is stable. For the incidence rate of the form g(I)h(S), we prove global stability, constructing a Lyapunov function and using the direct Lyapunov method. It is remarkable that the system dynamics is independent of how the incidence rate depends on the number of susceptible individuals. We demonstrate this result using a SIRS model and a SEIRS model as case studies. For other compartment epidemic models, the analysis is quite similar, and the same conclusion, namely stability of the equilibrium states, holds. PMID- 15778335 TI - Multiple modes of interaction between Lck and CD28. PMID- 15778336 TI - Proving negative selection in the thymus. PMID- 15778338 TI - Preventing intolerance: the induction of nonresponsiveness to dietary and microbial antigens in the intestinal mucosa. AB - The gut-associated lymphoid tissue (GALT) is constantly exposed to a variety of Ags and must therefore decipher a large number of distinct signals at all times. Responding correctly to each set of signals is crucial. When the GALT receives signals from the intestinal flora or food Ags, it must induce a state of nonresponsiveness (mucosal tolerance). In contrast, when pathogenic bacteria invade the intestinal mucosa, it is necessary to elicit strong T and B cell responses. The GALT is therefore in the position of constantly fighting intolerance to food and the commensal flora while effectively battling infectious microbes. Determining precisely which type of response to generate in each case is key to the prevention of immune dysregulation and tissue damage. PMID- 15778339 TI - Cutting edge: KIR2DL4 transduces signals into human NK cells through association with the Fc receptor gamma protein. AB - KIR2DL4 (2DL4, CD158d), a member of the human killer cell Ig-like receptor (KIR) family, triggers potent IFN-gamma responses but weak cytotoxicity in resting NK cells. 2DL4 mRNA has been detected in most NK cell clones from most humans examined, but surface protein expression is detectable only on CD56(high) NK cells from certain donors. The receptor possesses a transmembrane arginine residue, suggesting association with a signaling accessory protein that has remained elusive. We provide biochemical and functional evidence that FcepsilonRI gamma (gamma) associates with 2DL4 to promote surface expression and provide signal transducing function. Weak cytolytic responses triggered through 2DL4 may result from low stoichiometric association with gamma. Selective association with gamma distinguishes 2DL4 from all other activating forms of the KIR family, which alternatively associate with DNAX-activating protein (DAP)12. PMID- 15778340 TI - Cutting edge: IFN-gamma signaling to macrophages is required for optimal Valpha14i NK T/NK cell cross-talk. AB - Activated NK T cells are known to rapidly stimulate NK cells and, subsequently, CD8(+) T cells and B cells. In this report, we first demonstrate that the downstream effects induced by alpha-galactosylceramide activated NK T cells on NK cells are mainly dependent on IFN-gamma. We found that NK T cell activation of NK cells requires a functional IFN-gamma signaling in macrophages and dendritic cells but not in B cells, NK cells, or NK T cells. NK T cell activation is dendritic cell-dependent whereas NK T cell activation of NK cells is indirect and in part mediated by macrophages. Interestingly, in this context, macrophage participation in the CD1d Ag presentation of alpha-galactosylceramide to NK T cells is not necessary. These data indicate that NK T cell-dependent activation of macrophages is required for optimal NK T cell-induced stimulation of NK cells. PMID- 15778341 TI - Simultaneous LFA-1 and CD40 ligand antagonism prevents airway remodeling in orthotopic airway transplantation: implications for the role of respiratory epithelium as a modulator of fibrosis. AB - Airway remodeling is a prominent feature of certain immune-mediated lung diseases such as asthma and chronic lung transplant rejection. Under conditions of airway inflammation, the respiratory epithelium may serve an important role in this remodeling process. Given the proposed role of respiratory epithelium in nonspecific injury models, we investigated the respiratory epithelium in an immune-specific orthotopic airway transplant model. MHC-mismatched tracheal transplants in mice were used to generate alloimmune-mediated airway lesions. Attenuation of this immune injury and alteration of antidonor reactivity were achieved by the administration of combined anti-LFA-1/anti-CD40L mAbs. By contrast, without immunotherapy, transplanted airways remodeled with a flattening of respiratory epithelium and significant subepithelial fibrosis. Unopposed alloimmune injury for 10 days was associated with subsequent epithelial transformation and subepithelial fibrosis that could not be reversed with immunotherapy. The relining of donor airways with recipient-derived epithelium was delayed with immunotherapy resulting in partially chimeric airways by 28 days. Partial chimerism was sufficient to prevent luminal fibrosis. However, epithelial chimerism was also associated with airway remodeling. Therefore, there appears to be an intimate relationship between the morphology and level of chimerism of the respiratory epithelium and the degree of airway remodeling following alloimmune injury. PMID- 15778342 TI - Extracellular ATP induces cytokine expression and apoptosis through P2X7 receptor in murine mast cells. AB - Extracellular ATP and other nucleotides act through specific cell surface receptors and regulate a wide variety of cellular responses in many cell types and tissues. In this study, we demonstrate that murine mast cells express several P2Y and P2X receptor subtypes including P2X(7), and describe functional responses of these cells to extracellular ATP. Stimulation of bone marrow-derived mast cells (BMMC), as well as MC/9 and P815 mast cell lines with millimolar concentrations of ATP, resulted in Ca(2+) influx across the cellular membrane and cell permeabilization. Moreover, brief exposures to ATP were sufficient to induce apoptosis in BMMCs, MC/9, and P815 cells which involved activation of caspase-3 and -8. However, in the time period between commitment to apoptosis and actual cell death, ATP triggered rapid but transient phosphorylation of multiple signaling molecules in BMMCs and MC/9 cells, including ERK, Jak2, and STAT6. In addition, ATP stimulation enhanced the expression of several proinflammatory cytokines, such as IL-4, IL-6, IL-13, and TNF-alpha. The effects of ATP were mimicked by submillimolar concentrations of 3-O-(4'-benzoyl)-benzoyl-benzoyl-ATP, and were inhibited by pretreatment of mast cells with a selective blocker of human and mouse P2X(7) receptor, 1[N,O-bis(5-isoquinolinesulphonyl)-N-methyl-l tyrosyl]-4-phenylpiperazine, as well as oxidized ATP. The nucleotide selectivity and pharmacological profile data support the role for P2X(7) receptor as the mediator of the ATP-induced responses. Given the importance of mast cells in diverse pathological conditions, the ability of extracellular ATP to induce the P2X(7)-mediated apoptosis in these cells may facilitate the development of new strategies to modulate mast cell activities. PMID- 15778343 TI - Mice deficient in OX40 and CD30 signals lack memory antibody responses because of deficient CD4 T cell memory. AB - Recently, we reported that a CD4(+)CD3(-)CD11c(-) accessory cell provided OX40 dependent survival signals to follicular T cells. These accessory cells express both OX40 ligand and CD30 ligand, and the receptors, OX40 and CD30, are both expressed on Th2-primed CD4 T cells. OX40 and CD30 signals share common signaling pathways, suggesting that CD30 signals might substantially compensate in OX40 deficient mice. In this report we have dissected the signaling roles of CD30 alone and in combination with OX40. CD30-deficient mice showed an impaired capacity to sustain follicular germinal center responses, and recall memory Ab responses were substantially reduced. Deficiencies in OX40 and CD30 signals were additive; secondary Ab responses were ablated in double-deficient mice. Although the initial proliferation of OX40/CD30 double-knockout OTII transgenic T cells was comparable to that of their normal counterparts, they failed to survive in vivo, and this was associated with reduced T cell numbers associated with CD4(+)CD3(-) cells in B follicles. Finally, we show that OX40/CD30 double knockout OTII transgenic T cells fail to survive compared with normal T cells when cocultured with CD4(+)CD3(-) cells in vitro. PMID- 15778344 TI - Transgenic Ly-49A inhibits antigen-driven T cell activation and delays diabetes. AB - Activation of islet-specific T cells plays a significant role in the development of type 1 diabetes. In an effort to control T cell activation, we expressed the inhibitory receptor, Ly-49A, on islet-specific mouse CD4 cells. Ag-mediated activation of Ly-49A T cells was inhibited in vitro when the Ly-49A ligand, H 2D(d), was present on APCs. Ag-driven T cell proliferation, cytokine production, and changes in surface receptor expression were significantly reduced. Inhibition was also evident during secondary antigenic challenge. Addition of exogenous IL-2 did not rescue cells from inhibition, suggesting that Ly-49A engagement does not lead to T cell anergy. Importantly, in an adoptive transfer model, Ly-49A significantly delays the onset of diabetes. Together these results demonstrate that the inhibitory receptor Ly-49A effectively limits Ag-specific CD4 cell responses even in the presence of sustained autoantigen expression in vivo. PMID- 15778345 TI - IL-12 induction of mRNA encoding substance P in murine macrophages from the spleen and sites of inflammation. AB - Substance P (SP), a neuropeptide, interacts with the neurokinin 1 receptor (NK 1R) on immune cells to help control IFN-gamma production. In murine schistosomiasis mansoni, schistosome worms produce ova that incite focal Th2-type granulomatous inflammation within the liver and intestines. Normal gut is characterized by a controlled state of inflammation. IL-10 knockout mice develop chronic Th1-type colitis spontaneously. Both schistosome granulomas and gut mucosa display an SP immune regulatory circuit. However, the origin and regulation of SP production at these sites of inflammation are poorly understood. Macrophages are a potential source of SP. We therefore studied macrophages (F4/80(+)) from these models of inflammation. SP mRNA (preprotachykinin A (PPT A)) was detected within the schistosome granuloma, spleen, and lamina propria macrophages. Compared with those from wild-type mice, granuloma macrophages from STAT6(-/-) mice had 10-fold higher PPT A mRNA expression, whereas in STAT4(-/-) animals, PPT A mRNA expression was nearly abolished. IL-12 signals via STAT4 to induce Th1-type inflammation. It was demonstrated that IL-12, but not IL-18, induces SP mRNA expression in resting splenic macrophages from Schistosoma infected mice and in wild-type lamina propria mononuclear cells. Thus, macrophages are a source for SP at these sites of chronic inflammation, and IL-12 and STAT4 are regulators of macrophage SP mRNA expression. PMID- 15778346 TI - Mechanisms controlling termination of V-J recombination at the TCRgamma locus: implications for allelic and isotypic exclusion of TCRgamma chains. AB - Analyses of Vgamma-Jgamma rearrangements producing the most commonly expressed TCRgamma chains in over 200 gammadelta TCR(+) thymocytes showed that assembly of TCRgamma V-region genes display properties of allelic exclusion. Moreover, introduction of functionally rearranged TCRgamma and delta transgenes results in a profound inhibition of endogenous TCRgamma rearrangements in progenitor cells. The extent of TCRgamma rearrangements in these cells is best explained by a model in which initiation of TCRgamma rearrangements at both alleles is asymmetric, occurs at different frequencies depending on the V or J segments involved, and is terminated upon production of a functional gammadelta TCR. Approximately 10% of the cells studied contained two functional TCRgamma chains involving different V and Jgamma gene segments, thus defining a certain degree of isotypic inclusion. However, these cells are isotypically excluded at the level of cell surface expression possibly due to pairing restrictions between different TCRgamma and delta chains. PMID- 15778347 TI - Priming of CTLs by lymphocytic choriomeningitis virus depends on dendritic cells. AB - Appropriate activation of naive CD8(+) T cells depends on the coordinated interaction of these cells with professional APC that present antigenic peptides in the context of MHC class I molecules. It is accepted that dendritic cells (DC) are efficient in activating naive T cells and are unique in their capacity to prime CD8(+) T cell responses against exogenous cell-associated Ags. Nevertheless, it is unclear whether epitopes, derived from endogenously synthesized proteins and presented by MHC class I molecules on the surface of other APC including B cells and macrophages, can activate naive CD8(+) T cells in vivo. By infecting transgenic CD11c-DTR/GFP mice that allow conditional depletion of DC with lymphocytic choriomeningitis virus (LCMV), which infects all types of APC and elicits a vigorous CTL response, we unambiguously show that priming of LCMV-specific CD8(+) T cells is crucially dependent on DC, despite ample presence of LCMV-infected macrophages and B cells in secondary lymphoid organs. PMID- 15778348 TI - Stat3 activity in melanoma cells affects migration of immune effector cells and nitric oxide-mediated antitumor effects. AB - Infiltration of immune effector cells in tumors is critical for antitumor immune responses. However, what regulates immune cell infiltration of tumors remains to be identified. Stat3 is constitutively activated with high frequency in diverse cancers, promoting tumor cell growth and survival. Blocking Stat3 signaling in tumors in vivo results in tumor growth inhibition that involves killing of nontransfected tumor cells and infiltration of immune effector cells, suggesting that Stat3 activity in tumor cells might affect immune cell recruitment. However, dying tumor cells can also attract immune cells. In this study, we show in isogenic murine melanomas that natural Stat3 activity is associated with tumor growth and reduction of T cell infiltration. Blocking Stat3 signaling in the melanoma cells containing high Stat3 activity results in expression of multiple chemoattractants, leading to increased migration of lymphocytes, NK cells, neutrophils, and macrophages. In addition, blocking Stat3 triggers tumor cells to produce soluble factors capable of activating macrophage production of NO in vitro and in vivo. TNF-alpha and IFN-beta, which are secreted by Stat3-inhibited tumor cells, are able to activate macrophage NO production, whereas neutralizing TNF-alpha in the tumor supernatant from Stat3-blocked tumor cells abrogates nitrite production. Moreover, interrupting Stat3 signaling in tumor cells leads to macrophage-mediated, nitrite-dependent cytostatic activity against nontransduced tumor cells. These results suggest that tumor Stat3 activity affects recruitment of diverse immune effectors and it can be manipulated to activate the effector phase of innate immune responses. PMID- 15778349 TI - Anopheles mosquito bites activate cutaneous mast cells leading to a local inflammatory response and lymph node hyperplasia. AB - When Anopheles mosquitoes probe the skin for blood feeding, they inject saliva in dermal tissue. Mosquito saliva is known to exert various biological activities, but its perception by the immune system and its role in parasite transmission remain poorly understood. In the present study, we report on the cellular changes occurring in the mouse skin and draining lymph nodes after a Anopheles stephensi mosquito bite. We show that mosquito bites induce dermal mast cell degranulation, leading to fluid extravasation and neutrophil influx. This inflammatory response does not occur in mast cell-deficient W/W(v) mice, unless these are reconstituted specifically with mast cells. Mast cell activation caused by A. stephensi mosquito bites is followed by hyperplasia of the draining lymph node due to the accumulation of CD3(+), B220(+), CD11b(+), and CD11c(+) leukocytes. The T cell enrichment of the draining lymph nodes results from their sequestration from the circulation rather than local proliferation. These data demonstrate that mosquito bites and very likely saliva rapidly trigger the immune system, emphasizing the critical contribution of peripheral mast cells in inducing T cell and dendritic cell recruitment within draining lymph nodes, a prerequisite for the elicitation of T and B lymphocyte priming. PMID- 15778350 TI - Simultaneous induction of CD4 T cell tolerance and CD8 T cell immunity by semimature dendritic cells. AB - Previous studies suggested that depending on their maturation state, dendritic cells (DC) could either induce T cell tolerance (immature and semimature DC) or T cell activation (mature DC). Pretreatment of C57BL/6 mice with encephalitogenic myelin oligodendrocyte glycoprotein (MOG)(35-55) peptide-loaded semimature DC protected from MOG-induced autoimmune encephalomyelitis. This protection was mediated by IL-10-producing CD4 T cells specific for the self Ag. Here we show that semimature DC loaded with the MHC class II-restricted nonself peptide Ag (OVA) induce an identical regulatory T cell cytokine pattern. However, semimature DC loaded simultaneously with MHC class II- and MHC class I-restricted peptides, could efficiently initiate CD8 T cell responses leading to autoimmune diabetes in a TCR-transgenic adoptive transfer model. Double-peptide-loaded semimature DC also induced simultaneously in the same animal partially activated CD8 T cells with cytolytic function as well as protection from MOG-induced autoimmune encephalomyelitis. Our study suggests that the decision between tolerance and immunity not only depends on the DC, but also on the type and activation requirements of the responding T cell. PMID- 15778351 TI - IFN regulatory factor-2 cooperates with STAT1 to regulate transporter associated with antigen processing-1 promoter activity. AB - Class I MHC complexes (MHC(I)) are essential in mediating immune response. The transport of antigenic peptides (TAP) to MHC(I) and the stable expression of MHC(I) on the cell surface require the presence of a dedicated TAP. In this study we report that IFN-gamma and thrombopoietin (TPO) strongly increase TAP1 protein expression in megakaryocytes, followed by an enhanced expression of MHC(I) on the cell surface. This expression parallels the enhanced TAP1 promoter activity and TAP1 mRNA expression, which are independent of protein synthesis. We also show that this cytokine-dependent expression of TAP1 transcripts depends on STAT1 and IFN regulatory factor-2 (IRF-2), but not on IRF-1, and provide evidence that IRF 2 constitutively binds to the TAP1 gene promoter and enhances TAP1 promoter activity. We show that IRF-2 forms a complex with STAT1 and the cytokine responsive region of the TAP1 promoter in any TPO or IFN-gamma target cells tested. Interaction of IRF-2 and STAT1 on the promoter depends on the DNA-binding domain of IRF-2. Overall, our data indicate that TPO and IFN-gamma activate the expression of TAP1 via a new mechanism that involves functional cooperation between STAT1 and IRF-2 on the TAP1 promoter. PMID- 15778352 TI - Inducing P-selectin ligand formation in CD8 T cells: IL-2 and IL-12 are active in vitro but not required in vivo. AB - In vitro studies have demonstrated that IL-2 and IL-12 can support formation of P selectin ligands (P-SelL) in activated T cells, ligands that are variably required for efficient lymphocyte recruitment to sites of inflammation. To ascertain whether these cytokines were required for P-SelL formation in vivo, TCR transgenic CD8 T cells specific for male Ag (HY) were transferred into male mice under conditions in which either IL-2 and/or IL-15 or IL-12Rp40 were absent. P SelL formation at day 2 was unperturbed in HY-TCR IL-2(null) CD8 T cells responding in doubly deficient IL-2(null)IL-12(null) or IL-2(null)IL-15(null) male recipients. HY-specific CD8 T cell proliferative responses detected in both spleen and peritoneum occurred vigorously, but only splenic CD8 T cells up regulated P-SelL, demonstrating that in vivo induction of P-SelL is an active, nonprogrammed event following T cell activation and that despite the efficacy of IL-2 and IL-12 in supporting P-SelL formation in vitro, these cytokines appear to be dispensable for this purpose in vivo. PMID- 15778353 TI - Direct cross-priming by th lymphocytes generates memory cytotoxic T cell responses. AB - Under optimal Ag stimulation, CTL become functional effector and memory T cells. Professional APCs (pAPC) are considered essential for the activation of CTL, due to their unique capacity to provide costimulation and present exogenous Ags through MHC class I molecules. In this study, we report a novel means by which Th lymphocytes acquire and present MHC class I determinants to naive CTL. Although previous studies have looked at T cell Ag presentation to activated T cells, this study presents the first example of Ag presentation by Th cells to naive CTL. We report that activated Th cells can function as effective pAPC for CTL. Our results show that: 1) In addition to acquisition of cell surface molecules, including MHC class I/peptide complexes, from pAPC, Th cells can acquire and present MHC class I-binding peptides through TCR-MHC class II interactions with pAPC; 2) the acquired Ag can be functionally presented to CTL; and 3) Ag presentation by Th cells induces naive CTL to proliferate and preferentially differentiate into cells that phenotypically and functionally resemble central memory T cells. These findings suggest a novel role of Th cells as pAPC for the development of memory immune responses. PMID- 15778354 TI - Ex vivo characterization of the autoimmune T cell response in the HLA-DR1 mouse model of collagen-induced arthritis reveals long-term activation of type II collagen-specific cells and their presence in arthritic joints. AB - Although the pathogenesis of collagen-induced arthritis (CIA), a model of rheumatoid arthritis, is mediated by both collagen-specific CD4(+) T cells and Ab specific for type II collagen (CII), the role of CII-specific T cells in the pathogenesis of CIA remains unclear. Using tetrameric HLA-DR1 with a covalently bound immunodominant CII peptide, CII(259-273), we studied the development of the CII-specific T cell response in the periphery and arthritic joints of DR1 transgenic mice. Although the maximum number of DR1-CII-tetramer(+) cells was detected in draining lymph nodes 10 days postimmunization, these T cells accounted for only 1% or less of the CD4(+) population. After day 10, their numbers gradually decreased, but were still detectable on day 130. Examination of TCR expression and changes in CD62L, CD44(high), and CD69 expression by these T cells indicated that they expressed a limited TCR-BV repertoire and had clearly undergone activation. RT-PCR analysis of cytokine expression by the tetramer(+) T cells compared with tetramer(-) cells indicated the tetramer(+) cells expressed high levels of Th1 and proinflammatory cytokines, including IL-2, IFN-gamma, IL 6, TNF-alpha, and especially IL-17. Additionally, analysis of the synovium from arthritic paws indicated that the same CD4(+)/BV8(+)/BV14(+)/tetramer(+) T cells were present in the arthritic joints. These data demonstrate that although only small numbers of CII-specific T cells are generated during the development of CIA, these cells express very high levels of cytokine mRNA and appear to preferentially migrate to the arthritic joint, indicating a potential direct role of CII-specific T cells in the pathogenesis of CIA. PMID- 15778355 TI - Characterizing the impact of CD8 antibodies on class I MHC multimer binding. AB - Many studies have suggested that CD8 Abs affect the binding of class I MHC tetramers/multimers to CD8(+) T cells, which has led to the interpretation that CD8 participates directly in multimer binding. In contrast, a recent publication has argued that CD8 Abs instead cause reorganization of TCR distribution and hence have an indirect effect on multimer binding to the TCR alone. We address these issues by testing the role of CD8 and the impact of CD8 Abs on the binding of normal and mutant multimers to Ag-specific mouse T cells. Our data suggest that, in this system, CD8 Abs act directly on CD8 and only mediate their effects on multimer binding when CD8 is capable of binding to the multimer. These data reinforce the paradigm that CD8 plays an active and direct role in binding of class I MHC multimers. PMID- 15778356 TI - IL-12 or IL-4 prime human NK cells to mediate functionally divergent interactions with dendritic cells or tumors. AB - In the course of inflammatory responses in peripheral tissues, NK cells may be exposed to cytokines such as IL-12 and IL-4 released by other cell types that may influence their functional activities. In the present study we comparatively analyzed purified human peripheral blood NK cells that had been exposed to either IL-12 or IL-4 during short (overnight) incubation. We show that although IL-12 cultured NK cells produced abundant IFN-gamma, TNF-alpha, and GM-CSF in response to stimuli acting on the NKp46-activating receptor, IL-4-cultured NK cells did not release detectable levels of these cytokines. In contrast, IL-4-cultured NK cells produced significant levels of TNF-alpha and GM-CSF only when stimulated with PMA and ionomycin. In no instance could the production of IL-5 and IL-13 be detected. Importantly, IL-12-cultured, but not IL-4-cultured, NK cells displayed strong cytolytic activity against various tumor cells or immature dendritic cells (DCs). Moreover, only NK cells that had been cultured in IL-12 were able to induce substantial DC maturation. Our data suggest that NK cells exposed to IL-12 for a time interval compatible with in vivo responses may favor the selection of appropriate mature DCs for subsequent Th1 cell priming in secondary lymphoid organs. On the contrary, NK cells exposed to IL-4 do not exert DC selection, may impair efficient Th1 priming, and favor either tolerogenic or Th2-type responses. PMID- 15778358 TI - T cells in G1 provide a memory-like response to secondary stimulation. AB - The commitment of naive T cells to proliferate is a function of the strength and duration of stimuli mediated by the TCR and coreceptors. Ranges of 2-20 h of stimulation have been reported as necessary in vitro. Whether T cells actually experience uninterrupted stimulation for such long periods under physiological conditions is controversial. Here we ask whether commitment to proliferate requires continuous stimulation, or can T cells integrate intermittent periods of stimulation. T cells were stimulated for two short-term (subthreshold) periods (5 7 h) either sequentially or separated by an interval of rest. Naive lymph node T cells were able to integrate interrupted stimulation, even when the duration of rest was as long as 2 days. Furthermore, when short-term-stimulated T cells were separated by density, three populations were observed: low density blasts, intermediate density G(1) cells, and high density G(0) cells. Low density cells progressed to division without further stimulation, whereas G(0) and G(1) cells remained undivided. However, after a period of rest, a second subthreshold stimulation caused the G(1) but not the G(0) fraction to quickly proceed through the cell cycle. We conclude that noncycling T cells in the G(1) phase of the cell cycle remain in a state of readiness for prolonged periods of time, and may represent a population of memory-like effectors capable of responding rapidly to antigenic challenge. PMID- 15778357 TI - Effector CD4+ T cells generate intermediate caspase activity and cleavage of caspase-8 substrates. AB - Caspase-8 activation promotes cell apoptosis but is also essential for T cell activation. The extent of caspase activation and substrate cleavage in these divergent processes remains unclear. We show that murine effector CD4(+) T cells generated levels of caspase activity intermediate between unstimulated T cells and apoptotic populations. Both caspase-8 and caspase-3 were partially activated in effector T cells, which was reflected in cleavage of the caspase-8 substrates, c-FLIP(L), receptor interacting protein 1, and to a lesser extent Bid, but not the caspase-3 substrate inhibitor of caspase-activated DNase. Th2 effector CD4(+) T cells manifested more caspase activity than did Th1 effectors, and caspase blockade greatly decreased initiation of cell cycling. The current findings define the level of caspase activity and substrates during initiation of T cell cycling. PMID- 15778359 TI - IL-15 activates telomerase and minimizes telomere loss and may preserve the replicative life span of memory CD8+ T cells in vitro. AB - The preservation of the replicative life span of memory CD8(+) T cells is vital for long-term immune protection. Although IL-15 plays a key role in the homeostasis of memory CD8(+) T cells, it is unknown whether IL-15 regulates the replicative life span of memory CD8(+) T cells. In this study, we report an analysis of telomerase expression and telomere length in human memory phenotype CD8(+) T cells maintained by IL-15 in vitro. We demonstrate that IL-15 is capable of activating telomerase in memory CD8(+) T cells via Jak3 and PI3K signaling pathways. Furthermore, IL-15 induces a sustained level of telomerase activity over long periods of time, and in turn minimizes telomere loss in memory CD8(+) T cells after substantial cell divisions. These findings suggest that IL-15 activates stable telomerase expression and compensates telomere loss in memory phenotype CD8(+) T cells, and that telomerase may play an important role in memory CD8(+) T cell homeostasis. PMID- 15778360 TI - TRAIL-transduced dendritic cells protect mice from acute graft-versus-host disease and leukemia relapse. AB - TRAIL preferentially induces apoptotic cell death in a wide variety of transformed cells, whereas it induces no apoptosis, but inhibits activation of Ag specific T cells via blockade of cell cycle progression. Although accumulating results suggest that TRAIL is involved in the maintenance of immunological homeostasis under steady state conditions as well as in the initiation and progression of immunopathologies, the potential regulatory effect of TRAIL on immune responses and its therapeutic potential in immunological diseases remains unclear. We report in this study the potential usefulness of TRAIL-transduced dendritic cells (DCs) for the treatment of lethal acute graft-vs-host disease (GVHD) and leukemia relapse. DCs genetically modified to express TRAIL showed potent cytotoxicity against both alloreactive T cells and leukemic cells through the induction of apoptosis. In addition, treatment with genetically modified DCs expressing TRAIL of allogeneic BM transplants recipients with leukemia was effective for protection against acute GVHD and leukemia relapse. Thus, gene transfer of TRAIL to DCs is a novel modality for the treatment of acute GVHD and leukemia relapse by selective targeting of pathogenic T cells and leukemic cells. PMID- 15778361 TI - Increased expression of CD27 on activated human memory B cells correlates with their commitment to the plasma cell lineage. AB - Plasma cells (PC) or Ig-secreting cells (ISC) are terminally differentiated B cells responsible for the production of protective Ig. ISC can be generated in vitro by culturing human B cells with the T cell-derived stimuli CD40L, IL-2, and IL-10. ISC have traditionally been identified by the increased expression of CD38, analogous to primary human PC, and the acquired ability to secrete Ig. By tracking the proliferation history of activated B cells, we previously reported that the differentiation of memory B cells into CD38(+) B cells is IL-10 dependent, and increases in frequency with cell division. However, <50% of CD38(+) cells secreted Ig, and there was a population of CD38(-) ISC. Thus, the PC phenotype of CD38(+) cells generated in vitro did not correlate with PC function. To address this, we have examined cultures of activated memory B cells to accurately identify the phenotype of ISC generated in vitro. We found that CD27 is also up-regulated on memory B cells in an IL-10-dependent and division dependent manner, and that ISC segregated into the CD27(high) subset of activated memory B cells irrespective of the acquired expression of CD38. The ISC generated in these cultures expressed elevated levels of the transcription factors Blimp-1 and X box-binding protein-1 and reduced levels of Pax-5, and exhibited selective migration toward CXCL12, similar to primary PC. We propose that the differentiation of memory B cells into PC involves a transitional stage characterized by a CD27(high)CD38(-) phenotype with the acquired ability to secrete high levels of Ig. PMID- 15778362 TI - Plasmacytoid dendritic cells control TLR7 sensitivity of naive B cells via type I IFN. AB - Detailed information of human B cell activation via TLR may lead to a better understanding of B cell involvement in autoimmunity and malignancy. In this study we identified a fundamental difference in the regulation of TLR7- and TLR9 mediated B cell stimulation: whereas the induction of polyclonal naive B cell proliferation by the TLR7 ligands resiquimod (R848) and loxoribine required the presence of plasmacytoid dendritic cells (PDCs), activation via the TLR9 ligand CpG was independent of PDCs. We found that PDC-derived type I IFN enhanced TLR7 sensitivity of B cells by selectively up-regulating TLR7 expression. In contrast the expression levels of TLR9 and of other TLRs studied remained unchanged. In the presence of type I IFN, TLR7 ligation triggered polyclonal B cell expansion and B cell differentiation toward Ig-producing plasma cells; notably, this occurred independently of T cell help and B cell Ag. Human B cells did not respond to ligands of other TLRs including TLR2, TLR4 and TLR6 with and without type I IFN. In conclusion, our results reveal a distinct regulation of TLR7 and TLR9 function in human B cells and highlight TLR7 and TLR9 as unique targets for therapeutic intervention in B cell-mediated immunity and disease. PMID- 15778363 TI - An alternate core 2 beta1,6-N-acetylglucosaminyltransferase selectively contributes to P-selectin ligand formation in activated CD8 T cells. AB - Core 2 beta1,6-N-acetylglucosaminyltransferase (C2GlcNAcT) synthesizes essential core 2 O-glycans on selectin ligands, which mediate cell-cell adhesion required for lymphocyte trafficking. Although gene-deletion studies have implicated C2GlcNAcT-I in controlling selectin ligand-mediated cell trafficking, little is known about the role of the two other core 2 isoenzymes, C2GlcNAcT-II and C2GlcNAcT-III. We show that C2GlcNAcT-I-independent P-selectin ligand formation occurs in activated C2GlcNAcT-I(null) CD8 T cells. These CD8 T cells were capable of rolling under shear flow on immobilized P-selectin in a P-selectin glycoprotein ligand 1-dependent manner. RT-PCR analysis identified significant levels of C2GlcNAcT-III RNA, identifying this enzyme as a possible source of core 2 enzyme activity. Up-regulation of P-selectin ligand correlated with altered cell surface binding of the core 2-sensitive mAb 1B11, indicating that CD43 and CD45 are also physiological targets for this alternate C2GlcNAcT enzyme. Furthermore, C2GlcNAcT-I-independent P-selectin ligand induction was observed in an in vivo model. HY(tg) CD8 T cells from C2GlcNAcT-I(null) donors transferred into male recipients expressed P-selectin ligand in response to male Ag, although at reduced levels compared with wild-type HY(tg) CD8 T cells. Our data demonstrate that multiple C2GlcNAcT enzymes can contribute to P-selectin ligand formation and may cooperate with C2GlcNAcT-I in the control of CD8 T cell trafficking. PMID- 15778364 TI - CD40 engagement prevents peroxisome proliferator-activated receptor gamma agonist induced apoptosis of B lymphocytes and B lymphoma cells by an NF-kappaB-dependent mechanism. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a transcription factor important in fat metabolism and is emerging as an important regulator of immunity and inflammation. We previously demonstrated that normal and malignant B lineage cells express PPARgamma and die by apoptosis after PPARgamma agonist exposure. In this study, we used the WEHI-231 mouse B lymphoma and normal mouse spleen B lymphocytes to elucidate the mechanism of PPARgamma agonist-induced apoptosis, and to determine whether an apoptosis rescue mechanism exists. In WEHI 231 cells, the natural PPARgamma agonist 15-deoxy-Delta(12,14)-PGJ(2) and the synthetic PPARgamma agonist ciglitazone induced activation of caspase 3 and caspase 9, a decrease in mitochondrial membrane potential, and caused cleavage of the caspase substrate poly(ADP-ribose) polymerase. We next tested whether CD40, whose engagement delivers a potent prosurvival signal for B cells, could protect B cells from PPARgamma agonist-induced apoptosis. CD40 engagement with CD40L significantly blunted the ability of PPARgamma agonists to induce apoptosis of B lymphocytes and prevented the inhibition of NF-kappaB mobilization by 15-deoxy Delta(12,14)-PGJ(2) and ciglitazone. Interestingly, PPARgamma agonists induced an increase in IkappaBalpha and IkappaBbeta protein levels, which was prevented with CD40 engagement. The rescue mechanism induced by CD40 engagement was dependent on NF-kappaB, as an NF-kappaB inhibitor prevented rescue. Apoptosis induction by PPARgamma ligands may be important for immune regulation by killing B lymphocytes as a rapid means to dampen inflammation. Moreover, the ability of PPARgamma agonists to kill malignant B lineage cells has implications for their use as anti B lymphoma agents. PMID- 15778365 TI - The chemokine receptor CCR7 activates in dendritic cells two signaling modules that independently regulate chemotaxis and migratory speed. AB - CCR7 is necessary to direct dendritic cells (DCs) to secondary lymphoid nodes and to elicit an adaptative immune response. Despite its importance, little is known about the molecular mechanisms used by CCR7 to direct DCs to lymph nodes. In addition to chemotaxis, CCR7 regulates the migratory speed of DCs. We investigated the intracellular pathways that regulate CCR7-dependent chemotaxis and migratory speed. We found that CCR7 induced a G(i)-dependent activation of MAPK members ERK1/2, JNK, and p38, with ERK1/2 and p38 controlling JNK. MAPK members regulated chemotaxis, but not the migratory speed, of DCs. CCR7 induced activation of PI3K/Akt; however, these enzymes did not regulate either chemotaxis or the speed of DCs. CCR7 also induced activation of the GTPase Rho, the tyrosine kinase Pyk2, and inactivation of cofilin. Pyk2 activation was independent of G(i) and Src and was dependent on Rho. Interference with Rho or Pyk2 inhibited cofilin inactivation and the migratory speed of DCs, but did not affect chemotaxis. Interference with Rho/Pyk2/cofilin inhibited DC migratory speed even in the absence of chemokines, suggesting that this module controls the speed of DCs and that CCR7, by activating its components, induces an increase in migratory speed. Therefore, CCR7 activates two independent signaling modules, one involving G(i) and a hierarchy of MAPK family members and another involving Rho/Pyk2/cofilin, which control, respectively, chemotaxis and the migratory speed of DCs. The use of independent signaling modules to control chemotaxis and speed can contribute to regulate the chemotactic effects of CCR7. PMID- 15778366 TI - Impaired post-thymic development of regulatory CD4+25+ T cells contributes to diabetes pathogenesis in BB rats. AB - One of the BB rat diabetes (diabetes mellitus (DM)) susceptibility genes is an Ian5 mutation resulting in premature apoptosis of naive T cells. Impaired differentiation of regulatory T cells has been suggested as one possible mechanism through which this mutation contributes to antipancreatic autoimmunity. Using Ian5 congenic inbred rats (wild-type (non-lyp BB) and mutated (BB)), we assessed the development of BB regulatory CD8(-)4(+)25(+)T cells and their role in the pathogenesis of DM. BB rats have normal numbers of functional CD8( )4(+)25(+)Foxp3(+) thymocytes. The proportion of CD25(+) cells among CD8(-)4(+) recent thymic emigrants is also normal while it is increased among more mature CD8(-)4(+) T cells. However, BB CD8(-)4(+)25(+)Foxp3(+) thymocytes fail to undergo homeostatic expansion and survive upon transfer to nude BB rats while Foxp3 expression is reduced in mature CD8(-)4(+)25(+) T cells suggesting that these cells are mostly activated cells. Consistent with this interpretation, peripheral BB CD8(-)4(+)25(+) T cells do not suppress anti-TCR-mediated activation of non-lyp BB CD8(-)4(+)25(-) T cells but rather stimulate it. Furthermore, adoptive transfer of unfractionated T cells from diabetic BB donors induces DM in 71% of the recipients while no DM occurred when donor T cells are depleted of CD8(-)4(+)25(+) cells. Adoptive transfer of 10(6) regulatory non-lyp BB CD8(-)4(+)25(+) T cells to young BB rats protects the recipients from DM. Taken together, these results demonstrate that the BB rat Ian5 mutation alters the survival and function of regulatory CD8(-)4(+)25(+) T cells at the post thymic level, resulting in clonal expansion of diabetogenic T cells among peripheral CD8(-)4(+)25(+) cells. PMID- 15778367 TI - Cross-linking of CD45 on suppressive/regulatory T cells leads to the abrogation of their suppressive activity in vitro. AB - CD4(+)CD25(+) T cells have immunoregulatory and suppressive functions and are responsible for suppressing self-reactive cells and maintaining self-tolerance. In addition to CD4(+)CD25(+) T cells, there is some evidence that a fraction of CD4(+)CD25(-) T cells exhibit suppressive activity in vitro or in vivo. We have shown, using aged mice, that aging not only leads to a decline in the ability to mount CD4(+)CD25(-) T cell responses, but, at the same time, renders aged CD4(+)CD25(-) T cells suppressive. In this study we report two newly established mAbs that could abrogate the suppressive function of aged CD4(+)CD25(-) T cells. These mAbs recognized the same protein, the transmembrane phosphatase CD45. Cross linking of CD45 on aged CD4(+)CD25(-) T cells was required for the disruption of their suppressive activity. Surprisingly, these mAbs also abrogated the suppressive action of CD4(+)CD25(+) T cells in vitro. Our results demonstrate an unexpected function of CD45 as a negative regulator neutralizing the suppressive activity of aged CD4(+)CD25(-) and young CD4(+)CD25(+) T cells. PMID- 15778368 TI - Peripheral deletion of antigen-specific T cells leads to long-term tolerance mediated by CD8+ cytotoxic cells. AB - Peripheral deletion is one mechanism by which potentially self-reactive clones are removed whether they escape thymic deletion. We have examined the consequences of deleting Ag-specific T cells by i.v. injection of soluble Ag. Deletion of DO11.10 T cells by peptide was mediated predominately via a Fas/FasL mechanism. Animals that underwent deletion were tolerant to subsequent immunization with Ag, even when tolerant mice were given fresh Ag-specific DO11.10 T cells before immunization. Tolerance was mediated by CD8(+) T cells that killed the DO11.10-transgenic T cells in vivo. These data demonstrate that the programmed cell death of large numbers of T cells leads to peripheral tolerance mediated by CD8(+) CTLs. PMID- 15778369 TI - Stat5a inhibits IL-12-induced Th1 cell differentiation through the induction of suppressor of cytokine signaling 3 expression. AB - In previous studies, we have shown that Th2 cell differentiation is diminished but Th1 cell differentiation is increased in Stat5a-deficient (Stat5a(-/-)) CD4(+) T cells. In the present study, we clarified the molecular mechanisms of Stat5a-mediated Th cell differentiation. We found that enhanced Th1 cell differentiation and the resultant IFN-gamma production played a dominant inhibitory role in the down-regulation of IL-4-induced Th2 cell differentiation of Stat5a(-/-) CD4(+) T cells. We also found that IL-12-induced Stat4 phosphorylation and Th1 cell differentiation were augmented in Stat5a(-/-) CD4(+) T cells. Importantly, the expression of suppressor of cytokine signaling (SOCS)3, a potent inhibitor of IL-12-induced Stat4 activation, was decreased in Stat5a(-/ ) CD4(+) T cells. Moreover, a reporter assay showed that a constitutively active form of Stat5a but not Stat6 activated the SOCS3 promoter. Furthermore, chromatin immunoprecipitation assays revealed that Stat5a binds to the SOCS3 promoter in CD4(+) T cells. Finally, the retrovirus-mediated expression of SOCS3 restored the impaired Th cell differentiation of Stat5a(-/-) CD4(+) T cells. These results suggest that Stat5a forces the Th1/Th2 balance toward a Th2-type by preventing IL 12-induced Th1 cell differentiation through the induction of SOCS3. PMID- 15778370 TI - Receptors and counterreceptors involved in NK-B cell interactions. AB - In addition to the well-documented effect of NK cells on B cell differentiation via their ability to secrete IFN-gamma, NK cells can also induce, via direct cell cell interactions, germline transcripts (Igamma2a) necessary for switch recombination to IgG2a. Analysis of the ligand-receptor pairs that could be involved in this induction revealed that the expression of CD48 on B cells is crucial for the induction. NK cells from mice with targeted deletions of either the CD2 or the CD244 gene, both of which encode ligands for CD48, are compromised in their ability to induce B cell Igamma2a expression. Interestingly, although CD244 can bind to CD48 with a higher affinity, the ability of NK cells from CD244(-/-) mice to stimulate Igamma2a is not as compromised as NK cells from CD2( /-) mice. Despite the difference between cell surface receptors that are stimulated by NK cells vs those stimulated by the combination of LPS and IFN gamma, we show in this study that the initiation of gamma2a germline transcription is regulated by similar cis-acting elements located at the 3' end of the IgH locus. However, NK cells cannot induce the final steps of switch recombination resulting in the production of mature mRNA from recombined DNA. Our findings suggest that these different signaling pathways converge on regulatory elements that are common to germline transcription; however, because NK induction does not result in the final steps of switch recombination, some signals initiated by LPS plus IFN-gamma are not induced by NK cells. PMID- 15778371 TI - Galectin-1 acts as a soluble host factor that promotes HIV-1 infectivity through stabilization of virus attachment to host cells. AB - The establishment of HIV type 1 (HIV-1) infection is initiated by the stable attachment of the virion to the target cell surface. Although this process relies primarily upon interaction between virus-encoded gp120 and cell surface CD4, a number of distinct interactions influence binding of HIV-1 to host cells. In this study, we report that galectin-1, a dimeric beta-galactoside-binding protein, promotes infection with R5, X4, and R5X4 variants. Galectin-1 acts as a soluble adhesion molecule by facilitating attachment of HIV-1 to the cell surface. This postulate is based on experiments where galectin-1 rendered HIV-1 particles more refractory to various agents that block HIV-1 adsorption and coreceptor binding (i.e., a blocking anti-CD4, soluble CD4, human anti-HIV-1 polyclonal Abs; stromal cell-derived factor-1alpha; RANTES). Experiments performed with the fusion inhibitor T-20 confirmed that galectin-1 is primarily affecting HIV-1 attachment. The relevance of the present findings for the pathogenesis of HIV-1 infection is provided by the fact that galectin-1 is abundantly expressed in the thymus and lymph nodes, organs that represent major reservoirs for HIV-1. Moreover, galectin 1 is secreted by activated CD8(+) T lymphocytes, which are found in high numbers in HIV-1-positive patients. Therefore, it is proposed that galectin-1, which is released in an exocrine fashion at HIV-1 replication sites, can cross-link HIV-1 and target cells and promote a firmer adhesion of the virus to the cell surface, thereby augmenting the efficiency of the infection process. Overall, our findings suggest that galectin-1 might affect the pathogenesis of HIV-1 infection. PMID- 15778372 TI - Tumoricidal potential of native blood dendritic cells: direct tumor cell killing and activation of NK cell-mediated cytotoxicity. AB - Dendritic cells (DCs) are characterized by their unique capacity for primary T cell activation, providing the opportunity for DC-based cancer vaccination protocols. Novel findings reveal that besides their role as potent inducers of tumor-specific T cells, human DCs display additional antitumor effects. Most of these data were obtained with monocyte-derived DCs, whereas studies investigating native blood DCs are limited. In the present study, we analyze the tumoricidal capacity of M-DC8(+) DCs, which represent a major subpopulation of human blood DCs. We demonstrate that IFN-gamma-stimulated M-DC8(+) DCs lyse different tumor cell lines but not normal cells. In addition, we show that tumor cells markedly enhance the production of TNF-alpha by M-DC8(+) DCs via cell-to-cell contact and that this molecule essentially contributes to the killing activity of M-DC8(+) DCs. Furthermore, we illustrate the ability of M-DC8(+) DCs to promote proliferation, IFN-gamma production, and tumor-directed cytotoxicity of NK cells. The M-DC8(+) DC-mediated enhancement of the tumoricidal potential of NK cells is mainly dependent on cell-to-cell contact. These results reveal that, in addition to their crucial role in activating tumor-specific T cells, blood DCs exhibit direct tumor cell killing and enhance the tumoricidal activity of NK cells. These findings point to the pivotal role of DCs in triggering innate and adaptive immune responses against tumors. PMID- 15778373 TI - Three structurally and functionally divergent kinds of promoters regulate expression of clonally distributed killer cell Ig-like receptors (KIR), of KIR2DL4, and of KIR3DL3. AB - The generation of killer cell Ig-like receptor (KIR) expression patterns in NK cells involves variegated silencing of KIR genes by DNA methylation. To identify regulatory elements involved in KIR gene activation, upstream regions of KIR genes were functionally characterized in NK3.3 cells as well as in primary NK cells. Three kinds of KIR promoters were defined, controlling clonally expressed KIR genes, the constitutively active KIR2DL4, and the weakly expressed KIR3DL3. Upstream of a short core promoter common to all KIR genes, a region containing functionally divergent elements was characterized. Although this region had no impact on the activity of the KIR2DL3 promoter, an inhibitory element was identified in the KIR2DL4 promoter and an activating element was found in the KIR3DL3 promoter. Upon treatment with a methyltransferase inhibitor, KIR3DL3 expression could be readily induced showing that the low levels of KIR3DL3 expression in peripheral blood are due to sustained DNA methylation of an otherwise fully functional promoter. Analysis of transcription factor binding sites identified a functional acute myeloid leukemia (AML) site common to all three KIR promoters. Mutation of this site led to a substantial increase in activity of all KIR promoters. Among the different members of the AML family, AML 2 was identified as the predominant KIR binding factor. The present study suggests that AML-2 acts as a repressor of KIR expression in mature NK cells and opens the possibility that AML factors and associated cofactors are involved in regulation of KIR expression during NK cell development. PMID- 15778374 TI - Regulation of the murine Ddelta2 promoter by upstream stimulatory factor 1, Runx1, and c-Myb. AB - Accessibility control of V(D)J recombination at Ag receptor loci depends on the coordinate activities of transcriptional enhancers and germline promoters. Recombination of murine Tcrd gene segments is known to be regulated, at least in part, by the Tcrd enhancer (Edelta) situated in the Jdelta2-Cdelta intron. However, there has been little characterization of promoters and other cis-acting elements that are activated by or collaborate with Edelta and that might function to regulate Tcrd gene recombination events. We now describe a strong promoter that is tightly associated with the murine Ddelta2 gene segment. EMSAs reveal that upstream stimulatory factor 1, Runx1, c-Myb, lymphoid enhancer binding factor 1, NF1, and E47 all interact with this promoter in vitro. Of these, upstream stimulatory factor 1, Runx1, and c-Myb appear necessary for full promoter activity in transiently transfected cells. Moreover, the same three factors were found to interact with the promoter in vivo by chromatin immunoprecipitation. We suggest that these factors play important roles as Edelta dependent regulators of Ddelta2 accessibility in vivo. Consistent with the established roles of c-Myb and Runx factors in Edelta function, we detected low level, enhancer-independent activity of the Ddelta2 promoter in transient transfection experiments. We speculate that the Ddelta2 promoter may play a role as a weak, enhancer-independent regulator in vivo, and might contribute to residual Tcrd rearrangement in Edelta(-/-) mice. PMID- 15778375 TI - Plasticity and rigidity in adaptor protein-2-mediated internalization of the TCR:CD3 complex. AB - Many cell surface proteins are internalized via dileucine- or tyrosine-based motifs within their cytoplasmic domains by the heterotetrameric adaptor protein complex, AP-2. In this study we have examined how AP-2 mediates internalization of large cell surface receptors, such as the eight-chain TCR:CD3 complex. Although most receptors have a single signal that drives internalization, the TCR complex has two (D/E)xxxL(L/I) motifs and 20 YxxO motifs. Using 293T cells, we show that AP-2 is completely dependent on both signals to mediate TCR internalization, because deletion of either completely blocks this process. Significant plasticity and redundancy were observed in the use of the YxxO motifs, with a clear hierarchy in their use (CD3delta > CD3gamma >or= CD3zeta >> CD3epsilon). Remarkably, a single, membrane-distal YxxO motif in CD3delta could mediate approximately 75% of receptor internalization, whereas its removal only reduced internalization by approximately 20%. In contrast, significant rigidity was observed in use of the (D/E)xxxL(L/I) motif in CD3gamma. This was due to an absolute requirement for the position of this signal in the context of the TCR complex and for a highly conserved lysine residue, K128, which is not present in CD3delta. These contrasting requirements suggest a general principle by which AP 2 may mediate the internalization of large, multichain complexes. PMID- 15778376 TI - Stable activation of phosphatidylinositol 3-kinase in the T cell immunological synapse stimulates Akt signaling to FoxO1 nuclear exclusion and cell growth control. AB - We have previously reported at the single cell level that PI3K is activated after conjugate formation between T lymphocytes and APCs. However, in contrast to cells exposed to an asymmetrical signal that usually increase 3'-phosphoinositides (3' PI) transiently in the region of the activated receptors, T cells contacting APC accumulate 3'-PI across their whole plasma membrane far beyond the region of the immunological synapse (IS). Importantly, this effect is maintained over time, for hours, and although PI3K-dependent pathways translate in various cell types extracellular stimuli into a wide range of biological events, in primary T cells this stability is mostly required for cell division induced by Ag. Using imaging methodologies, the present article elucidates the molecular mechanisms responsible for this particular functioning of the PI3K pathway in primary human T lymphocytes interacting with APCs, especially with dendritic cells. The results reveal that the IS unremittingly recruits PI3K to maintain high 3'-PI levels in T cells through phosphotyrosine-dependent mechanisms, suggesting a major participation of class Ia PI3K. This persistent activation of PI3K results in the Akt-dependent sequestration of the FoxO transcription factor, FoxO1, outside the nucleus of T cells interacting with APCs. Using an active form of FoxO1, we demonstrate that this compartmentalization process can affect T cell growth after Ag recognition. We conclude that the need for sustained PI3K signaling within the consolidated IS is probably an undemanding tactic used by primary T cells critical for initiating cell cycle progression through the prolonged inactivation of FoxO1, one important factor that can control cell quiescence. PMID- 15778377 TI - Linking C5 deficiency to an exonic splicing enhancer mutation. AB - As an important component of the innate immune system, complement provides the initial response to prevent infections by pathogenic microorganisms. Patients with dysfunction of C5 display a propensity for severe recurrent infections. In this study, we present a patient with C5 deficiency demonstrated by immunochemical and functional analyses. Direct sequencing of all C5 exons displayed no mutation of obvious functional significance, except for an A to G transition in exon 10 predicting an exchange from lysine to arginine. This sequence alteration was present in only one allele of family members with a reduced serum C5 concentration and in both alleles of the patient with almost complete C5 deficiency, suggesting that this alteration may be producing the phenotype. Recent findings indicate that distinct nucleotide sequences, termed exonic splicing enhancers (ESEs), influence the splicing process. cDNA from all family members harboring the mutated allele showed skipping of exon 10, which resulted in a premature STOP codon, explaining the lack of C5 in the propositus. Sequence analysis of the mutated region revealed the substitution to be located within an ESE, as predicted by the RESCUE-ESE program. The altered ESE sequence is located close to the 5' splicing site and also lowers the predicted strength of the splice site itself. This apparently inconsequential sequence alteration represents a noncanonical splicing mutation altering an ESE. Our finding sheds a new light on the role of putative silent/conservative mutations in disease associated genes. PMID- 15778378 TI - p38 MAPK autophosphorylation drives macrophage IL-12 production during intracellular infection. AB - The intracellular protozoan Toxoplasma gondii triggers rapid MAPK activation in mouse macrophages (Mphi). We used synthetic inhibitors and dominant-negative Mphi mutants to demonstrate that T. gondii triggers IL-12 production in dependence upon p38 MAPK. Chemical inhibition of stress-activated protein kinase/JNK showed that this MAPK was also required for parasite-triggered IL-12 production. Examination of upstream MAPK kinases (MKK) 3, 4, and 6 that function as p38 MAPK activating kinases revealed that parasite infection activates only MKK3. Nevertheless, in MKK3(-/-) Mphi, p38 MAPK activation was near normal and IL-12 production was unaffected. Recently, MKK-independent p38alpha MAPK activation via autophosphorylation was described. Autophosphorylation depends upon p38alpha MAPK association with adaptor protein, TGF-beta-activated protein kinase 1-binding protein-1. We observed TGF-beta-activated protein kinase 1-binding protein-1 p38alpha MAPK association that closely paralleled p38 MAPK phosphorylation during Toxoplasma infection of Mphi. Furthermore, a synthetic p38 catalytic-site inhibitor blocked tachyzoite-induced p38alpha MAPK phosphorylation. These data are the first to demonstrate p38 MAPK autophosphorylation triggered by intracellular infection. PMID- 15778379 TI - Maintenance of pulmonary Th1 effector function in chronic tuberculosis requires persistent IL-12 production. AB - The mechanisms that prevent reactivation of latent Mycobacterium tuberculosis infection in asymptomatic individuals are poorly understood. Although IL-12 is critical for the induction of IFN-gamma-dependent host control of M. tuberculosis, the requirement for the cytokine in the maintenance of host resistance and pulmonary Th1 effector function has not yet been formally examined. In this study, we reconstituted IL-12p40-deficient mice with IL-12 during the first 4 wk of infection and then assessed the effects of cytokine withdrawal. Although IL-12 administration initially resulted in restricted mycobacterial growth and prolonged survival, the reconstituted animals eventually succumbed to infection. This breakdown in bacterial control was accompanied by a marked reduction in the numbers of IFN-gamma-producing CD4(+) T cells in lungs. Moreover, whereas CD4(+) T cells isolated from chronically infected wild-type mice expanded and transferred long-term protection to M. tuberculosis-challenged RAG(-/-) mice, they failed to do so in IL-12p40-deficient RAG(-/-) recipients and were clearly reduced in frequency within pulmonary granulomas in the latter animals. These studies establish that continuous IL-12 production is necessary for maintenance of the pulmonary Th1 cells required for host control of persistent M. tuberculosis infection and suggest that breakdown of this mechanism could be a contributing factor in reactivated disease. PMID- 15778380 TI - Smad3 deficiency in mast cells provides efficient host protection against acute septic peritonitis. AB - Mast cells play an important role in innate immunity as well as in allergic reaction. However, regulatory mechanisms underlying mast cell-mediated innate immune responses remain largely unknown. Here we determined whether Smad3, a major signal transducer of TGF-beta, regulates innate immune response by mast cells against Gram-negative bacteria. Bone marrow-derived mast cells (BMMC) obtained from Smad3 null mutant mice showed augmented capacity to produce proinflammatory cytokines upon stimulation with a Gram-negative bacteria associated product, LPS. In acute septic peritonitis model induced by cecal ligation and puncture, mast cell-deficient W/W(v) mice reconstituted with Smad3 null BMMC had significantly higher survival rate than W/W(v) mice reconstituted with wild-type BMMC, which was associated with higher production of proinflammatory cytokines in the peritoneal cavity. These in vitro and in vivo results suggest that Smad3 in mast cells functions as inhibitory for mast cell mediated innate immune response against Gram-negative bacteria. Suppression of Smad3 expression in mast cells may thus have therapeutic potential for Gram negative bacterial infection such as acute septic peritonitis by augmenting innate immune responses of mast cells. PMID- 15778381 TI - CD8-mediated protection against Ebola virus infection is perforin dependent. AB - CD8 T cells have been shown to play an important role in the clearance and protection against fatal Ebola virus infection. In this study, we examined the mechanisms by which CD8 T cells mediate this protection. Our data demonstrate that all normal mice infected s.c. with a mouse-adapted Ebola virus survived the infection, as did 100% of mice deficient in Fas and 90% of those deficient in IFN gamma. In contrast, perforin-deficient mice uniformly died after s.c. challenge. Perforin-deficient mice failed to clear viral infection even though they developed normal levels of neutralizing anti-Ebola Abs and 5- to 10-fold higher levels of IFN-gamma than control mice. Using MHC class I tetramers, we have also shown that perforin-deficient mice have 2- to 4-fold higher numbers of Ebola specific CD8s than control mice. These findings suggest that the clearance of Ebola virus is perforin-dependent and provide an additional example showing that this basic immunologic mechanism is not limited to the clearance of noncytopathic viruses. PMID- 15778382 TI - Inverse correlation of maturity and antibacterial activity in human dendritic cells. AB - Dendritic cells (DCs) are a key part of host defense against microbial pathogens, being part of the innate immune system, but also instructing the adaptive T cell response. This study was designed to evaluate whether human DCs directly contribute to innate immunity by killing intracellular bacteria, using tuberculosis as a model. DCs were detected in bronchoalveolar lavage samples indicating that DCs are available for immediate interaction with Mycobacterium tuberculosis (M. Tb) after inhalation of the pathogen. The phenotype of DC in bronchoalveolar lavage closely resembles monocyte-derived immature DC (iDC) according to the expression of CD1a, CD83, and CCR7. The antimicrobial activity of iDC against intracellular M. Tb inversely correlated with TNF-alpha-release and was enhanced by treatment with anti-TNF-alpha Abs. Differentiation of iDC into mature DC by addition of TNF-alpha or activation via Toll-like receptors further reduced killing of M. Tb. The antibacterial activity against intracellular M. Tb of all DCs was significantly lower than alveolar macrophages. Therefore, the maintenance of a pool of DCs at the site of disease activity in tuberculosis, and the maturation of these DC by TNF-alpha provides a mechanism by which M. Tb escapes the innate immune system. PMID- 15778383 TI - Cross-talk between CD14 and complement receptor 3 promotes phagocytosis of mycobacteria: regulation by phosphatidylinositol 3-kinase and cytohesin-1. AB - The glycosylphosphatidyl anchored molecule CD14 to the monocyte membrane plays a prominent role in innate immunity, and the paradigms for CD14 selective signaling are beginning to be elucidated. In this study, transfected human monocytic cell line THP-1 and Chinese hamster ovary (CHO) fibroblastic cells were used to examine phagocytosis of Mycobacterium bovis bacillus Calmette-Guerin (BCG). Flow cytometry was combined with molecular and biochemical approaches to demonstrate a dual mechanism for BCG internalization involving either CD14 alone or a CD14 regulated complement receptor (CR)3-dependent pathway. Phagocytosis by CD14 positive THP-1 cells was attenuated by phosphatidylinositol-3 inhibitors LY294002 and wortmannin and experiments using transfected CHO cells showed substantial accumulation of phosphatidylinositol-3,4,5-trisphosphate at the BCG attachment site in CHO cells expressing CD14 and TLR2 suggesting that bacteria bind to CD14 and use TLR2 to initiate a PI3K signaling pathway. Additional experiments using blocking Abs showed that anti-TLR2 Abs inhibit phagocytosis of BCG by THP-1 cells. Furthermore, knockdown of cytohesin-1, a PI3K-regulated adaptor molecule for beta(2) integrin activation, specifically abrogated CD14-regulated CR3 ingestion of BCG consistent with the observation of physical association between CR3 and cytohesin-1 in cells stimulated with mycobacterial surface components. These findings reveal that mycobacteria promote their uptake through a process of "inside-out" signaling involving CD14, TLR2, PI3K, and cytohesin-1. This converts low avidity CR3 into an active receptor leading to increased bacterial internalization. PMID- 15778384 TI - Uptake of granulysin via lipid rafts leads to lysis of intracellular Listeria innocua. AB - The bacteriolytic activity of CTL is mediated by granulysin, which has been reported to kill intracellular Mycobacterium tuberculosis in dendritic cells (DC) with high efficiency. Despite that crucial effector function, the killing mechanism and uptake of granulysin into target cells have not been well investigated. To this end we analyzed granulysin binding, uptake, and the subsequent lysis of intracellular Listeria innocua in human DC. Recombinant granulysin was found to be actively taken up by DC into early endosomal Ag 1 labeled endosomes, as detected by immunofluorescence. Further transfer to L. innocua-containing phagosomes was indicated by colocalization of bacterial DNA with granulysin. After uptake of granulysin by DC, lysis of L. innocua was found in a dose-dependent manner. Uptake as well as lysis of Listeria were inhibited after blocking endocytosis by lowering the temperature and by cholesterol depletion of DC. Colocalization of granulysin with cholera toxin during uptake showed binding to and internalization via lipid rafts. In contrast to cholera toxin, which was targeted to the perinuclear compartment, granulysin was found exclusively in endosomal-phagosomal vesicles. Lipid raft microdomains, enriched in the immunological synapse, may thus enhance uptake and transfer of granulysin into bacterial infected host cells. PMID- 15778385 TI - Early role of CD4+ Th1 cells and antibodies in HER-2 adenovirus vaccine protection against autochthonous mammary carcinomas. AB - HER-2 is an oncogenic tumor-associated Ag that is overexpressed in several human tumors including breast and ovarian cancer. The efficacy and mechanism of a HER-2 expressing recombinant adenoviral vaccine to protect against tumorigenesis was examined using HER-2 transgenic (BALB-neuT) mice, which develop spontaneous breast tumors in all 10 mammary glands, and also using a transplantable mouse tumor model. Vaccination beginning at 6-8 wk of age (through 19 wk of age) prevented development of spontaneous mammary tumors even after 50 wk, whereas the animals in the control groups had tumors in all mammary glands by 25 wk. Such long-term protection after the last boost has not been achieved previously in this transgenic mouse in which the oncogene is continuously spawning tumorigenesis. Using beta(2)-microglobulin-knockout, IFN-gamma-knockout, and B cell-deficient mice, CD4(+) and CD8(+) cell depletion, and Ab transfer studies, we show that induction of anti-HER-2/neu Abs are both necessary and sufficient for protection, and the IgG2a isotype is most effective. In contrast, CD8(+) T cells are not necessary at all, and CD4(+) T cells are necessary for only 36-48 h after immunization to provide help for B cells but not as effector cells. Equal protection in immunized mice deficient in FcgammaRI/III excluded an FcR-mediated mechanism. Anti-HER-2 serum not only inhibited growth of mammary tumor cell lines expressing HER-2 in vitro but also protected mice from tumors in vivo, suggesting a direct action of Ab on the tumor cells. Such a vaccine may provide Ab-mediated protection against HER-2-expressing breast cancers in humans. PMID- 15778386 TI - Infection biology of a novel alpha-crystallin of Mycobacterium tuberculosis: Acr2. AB - Heat shock proteins assist the survival of Mycobacterium tuberculosis (MTB) but also provide a signal to the immune response. The gene most strongly induced by heat shock in MTB is Rv0251c, which encodes Acr2, a novel member of the alpha crystallin family of molecular chaperones. The expression of acr2 increased within 1 h after infection of monocytes or macrophages, reaching a peak of 18- to 55-fold by 24 h. Inhibition of superoxide action reduced the intracellular increase in acr2. Despite this contribution to the stress response of MTB, the gene for acr2 appears dispensable; a deletion mutant (Deltaacr2) was unimpaired in log phase growth and persisted in IFN-gamma-activated human macrophages. Acr2 protein was strongly recognized by cattle with early primary Mycobacterium bovis infection and by healthy MTB-sensitized people. Within the latter group, those with recent exposure to infectious tuberculosis had, on average, 2.6 times the frequency of Acr2-specific IFN-gamma-secreting T cells than those with more remote exposure (p = 0.009). These data show that, by its up-regulation early after entry to cells, Acr2 gives away the presence of MTB to the immune response. The demonstration that there is infection stage-specific immunity to tuberculosis has implications for vaccine design. PMID- 15778387 TI - Yersinia outer protein P inhibits CD8 T cell priming in the mouse infection model. AB - Pathogenic yersiniae translocate a mixture of effector proteins called Yersinia outer proteins (Yops) into the cytosol of eukaryotic cells by their type III secretion system. YopP is one of the best characterized of these effector proteins and known to inhibit the proinflammatory response of the host by interfering with NF-kappaB signal transduction and inducing apoptosis of macrophages. The effects of YopP on the immune response were studied by a Yersinia Ag-independent approach using bacteria that translocate the well characterized model Ag listeriolysin O of Listeria monocytogenes via their type III secretion system. In this study we demonstrate a novel function for YopP in vivo. It is shown for the first time that YopP not only counteracts the innate immune defense but also inhibits the adaptive immune system by suppressing the development of an effective CD8 T cell response in a mouse model. A possible mechanism for this could be the inhibition of Ag presentation by dendritic cells (DC). In vitro this is shown to be due to the rapid induction of programmed DC death and to inhibition of DC maturation. Using this approach we could further show that the listeriolysin O-specific CD8 T cells generated in vivo by the yopP mutant are functional and are able to protect mice against a lethal challenge with wild type Listeria. PMID- 15778388 TI - Mastoparan, a G protein agonist peptide, differentially modulates TLR4- and TLR2 mediated signaling in human endothelial cells and murine macrophages. AB - Previous studies have implicated a role for heterotrimeric G protein-coupled signaling in B cells, monocytes, and macrophages stimulated with LPS and have shown that G proteins coimmunoprecipitate with membrane-bound CD14. In this study, we have extended these observations in human dermal microvessel endothelial cells (HMEC) that lack membrane-bound CD14 and in murine macrophages to define further the role of heterotrimeric G proteins in TLR signaling. Using the wasp venom-derived peptide, mastoparan, to disrupt G protein-coupled signaling, we identified a G protein-dependent signaling pathway in HMEC stimulated with TLR4 agonists that is necessary for the activation of p38 phosphorylation and kinase activity, NF-kappaB and IL-6 transactivation, and IL-6 secretion. In contrast, HMEC activation by TLR2 agonists, TNF-alpha, or IL-1beta was insensitive to mastoparan. In the murine macrophage cell line, RAW 264.7, and in primary murine macrophages, G protein dysregulation by mastoparan resulted in significant inhibition of LPS-induced signaling leading to both MyD88-dependent and MyD88-independent gene expression, while TLR2-mediated gene expression was not significantly inhibited. In addition to inhibition of TLR4-mediated MAPK phosphorylation in macrophages, mastoparan blunted IL-1R-associated kinase-1 kinase activity induced by LPS, but not by TLR2 agonists, yet failed to affect phosphorylation of Akt by phosphoinositol-3-kinase induced by either TLR2- or TLR4-mediated signaling. These data confirm the importance of heterotrimeric G proteins in TLR4-mediated responses in cells that use either soluble or membrane associated CD14 and reveal a level of TLR and signaling pathway specificity not previously appreciated. PMID- 15778389 TI - Induction of human T cell leukemia virus type I receptors on quiescent naive T lymphocytes by TGF-beta. AB - The retrovirus human T cell leukemia virus (HTLV) type I (HTLV-I) is primarily transmitted by breast-feeding or sexual contact, by cell-to-cell contact between T cells. TGF-beta, which has been shown to enhance transmission of HTLV-I in vitro, is found at high levels in breast milk and semen. In this study, the ability of TGF-beta to regulate expression of molecules involved in HTLV-I binding and entry was examined. Previous studies using a soluble form of the HTLV I envelope protein SU have shown that quiescent human T cells do not express cell surface molecules that specifically bind SU. After T cell activation, HTLV SU binding proteins are rapidly induced. In this study, we report that TGF-beta induces expression of proteins that bind soluble HTLV SU and HTLV virions on naive CD4(+) T lymphocytes. The induction of these proteins occurred without cell cycle entry or expression of activation markers, involved TGF-beta-induced intracellular signaling, and required de novo transcription and translation. Treatment of naive CD4(+) T lymphocytes with TGF-beta induced expression of GLUT 1, which has recently been reported to function as a receptor for HTLV. Treatment of a TGF-beta-sensitive human myeloid cell line increased the titer of both HTLV I- and HTLV-II-pseudotyped viruses. Although earlier studies suggested that HTLV SU binding proteins might be an early marker of T cell activation and/or cell proliferation, we report in this study that TGF-beta induces binding of HTLV virions and expression of glucose transporter type 1 in primary CD4(+) T lymphocytes that remain quiescent. PMID- 15778390 TI - Structure-function relationships among human cathelicidin peptides: dissociation of antimicrobial properties from host immunostimulatory activities. AB - Cathelicidins and other antimicrobial peptides are deployed at epithelial surfaces to defend against infection. These molecules have broad-spectrum killing activity against microbes and can have effects on specific mammalian cell types, potentially stimulating additional immune defense through direct chemotactic activity or induction of cytokine release. In humans, the cathelicidin hCAP18/LL 37 is processed to LL-37 in neutrophils, but on skin it can be further proteolytically processed to shorter forms. The influence of these cathelicidin peptides on keratinocyte function is not known. In the current study, DNA microarray analysis and confirmatory protein analysis showed that LL-37 affects the expression of several chemokines and cytokines by keratinocytes. Analysis of a synthetic peptide library derived from LL-37 showed that antimicrobial activity against bacterial, fungal, and viral skin pathogens resides within specific domains of the parent peptide, but antimicrobial activity does not directly correlate with the ability to stimulate IL-8 production in keratinocytes. IL-8 release was induced by d- and l-amino acid forms of cathelicidin and correlated with membrane permeability, suggesting that highly structure-specific binding to a cell surface receptor is not likely. However, this effect was inhibited by either pertussis toxin or AG1478, an epidermal growth factor receptor tyrosine kinase inhibitor, suggesting that cathelicidin may indirectly stimulate multiple signaling pathways associated with cell surface receptors. Taken together, these observations suggest that proteolytic processing may alter the balance between cathelicidin antimicrobial and host immunostimulatory functions. PMID- 15778391 TI - NADPH oxidase-derived reactive oxygen species-mediated activation of ERK1/2 is required for apoptosis of human neutrophils induced by Entamoeba histolytica. AB - The extracellular tissue penetrating protozoan parasite Entamoeba histolytica has been known to induce host cell apoptosis. However, the intracellular signaling mechanism used by the parasite to trigger apoptosis is poorly understood. In this study, we investigated the roles of reactive oxygen species (ROS), and of MAPKs in the Entamoeba-induced apoptosis of human neutrophils. The neutrophils incubated with live trophozoites of E. histolytica revealed a marked increase of receptor shedding of CD16 as well as phosphatidylserine (PS) externalization on the cell surface. The Entamoeba-induced apoptosis was effectively blocked by pretreatment of cells with diphenyleneiodonium chloride (DPI), a flavoprotein inhibitor of NADPH oxidase. A large amount of intracellular ROS was detected after exposure to viable trophozoites, and the treatment with DPI strongly inhibited the Entamoeba-induced ROS generation. However, a mitochondrial inhibitor rotenone did not attenuate the Entamoeba-induced ROS generation and apoptosis. Although E. histolytica strongly induced activation of ERK1/2 and p38 MAPK in neutrophils, the activation of ERK1/2 was closely associated with ROS mediated apoptosis. Pretreatment of neutrophils with MEK1 inhibitor PD98059, but not p38 MAPK inhibitor SB202190, prevented Entamoeba-induced apoptosis. Moreover, DPI almost completely inhibited Entamoeba-induced phosphorylation of ERK1/2, but not phosphorylation of p38 MAPK. These results strongly suggest that NADPH oxidase-derived ROS-mediated activation of ERK1/2 is required for the Entamoeba induced neutrophil apoptosis. PMID- 15778392 TI - IFN-alpha enhances TLR3-mediated antiviral cytokine expression in human endothelial and epithelial cells by up-regulating TLR3 expression. AB - TLRs play a critical role in early innate immune response to virus infection. TLR3 together with TLR7 and TLR8 constitute a powerful system to detect genetic material of RNA viruses. TLR3 has been shown to bind viral dsRNA whereas TLR7 and TLR8 are receptors for viral single-stranded RNA. In this report we show that TLR7 or TLR8 are not expressed in human epithelial A549 cells or in HUVECs. Accordingly, A549 cells and HUVECs were unresponsive to TLR7/8 ligand R848. TLR3 was expressed at a higher level in HUVECs than in A549 cells. The TLR3 ligand poly(I:C) up-regulated IFN-beta, IL-28, IL-29, STAT1, and TLR3 expression in HUVECs but not in A549 cells. An enhanced TLR3 expression by transfection or by IFN-alpha stimulation conferred poly(I:C) responsiveness in A549 cells. Similarly, IFN-alpha pretreatment strongly enhanced poly(I:C)-induced activation of IFN-beta, IL-28, and IL-29 genes also in HUVECs. In conclusion, our results suggest that IFN-alpha-induced up-regulation of TLR3 expression is involved in dsRNA activated antiviral response in human epithelial and endothelial cells. PMID- 15778393 TI - Requirement for TLR9 in the immunomodulatory activity of Propionibacterium acnes. AB - Propionibacterium acnes (formerly Corynebacterium parvum) is part of the human flora and, as such, is associated with several human pathologies. It possesses strong immunomodulatory activities, which makes this bacterium interesting for prophylactic and therapeutic vaccination. The bacterial component(s) and the host receptor(s) involved in the induction of these activities are poorly understood. We show in this study that TLR9 is crucial in generating the characteristic effects of killed P. acnes priming in the spleen, such as extramedullary hemopoiesis and organ enlargement, and granuloma formation in the liver. Furthermore, the ability to overproduce TNF-alpha and IFN-gamma in response to LPS, lipid A, synthetic lipopeptide Pam(3)CysK(4), or whole killed bacteria was present in P. acnes-primed wild-type, but not TLR9(-/-), mice. Finally, P. acnes priming failed to induce enhanced resistance to murine typhoid fever in TLR9(-/-) mice. Thus, TLR9 plays an essential role in the induction of immunomodulatory effects by P. acnes. Because IFN-gamma is a key mediator of these effects, and enhanced IFN-gamma mRNA expression was absent in spleen and liver of P. acnes primed TLR9(-/-) mice, we conclude that TLR9 is required for the induction of IFN gamma by P. acnes. PMID- 15778394 TI - Regulation of p38 MAPK by MAPK kinases 3 and 6 in fibroblast-like synoviocytes. AB - The p38 MAPK signal transduction pathway is a key regulator of IL-1 and TNF-alpha production in rheumatoid arthritis. Previous studies demonstrated that upstream MAPK kinases (MKK3 and MKK6) that regulate p38 are activated in rheumatoid arthritis synovium. However, their functional relevance in fibroblast-like synoviocytes (FLS) has not been determined. To investigate the relative contribution of MKK3 and MKK6 to p38 activation, the effect of dominant-negative (DN) MKK3 and MKK6 constructs on cultured FLS was evaluated. Cultured FLS were stimulated with medium or IL-1beta, and immunoblotting was performed. In some experiments, cells were lysed and immunoprecipitated with anti-p38 Ab, followed by in vitro kinase assay with [gamma-(32)P]ATP and GST-activating transcription factor-2 as substrate. IL-1beta rapidly induced p38 phosphorylation in cells transfected with empty vector (pcDNA3.1), but was inhibited by 25% in cells expressing DN MKK3 or DN MKK6. Cotransfection with both DN plasmids decreased phospho-p38 by almost 75%. In vitro kinase assays on IL-1-stimulated FLS also showed that the combination of DN MKK3 and DN MKK6 markedly decreased kinase activity compared with empty vector or the individual DN plasmids. Furthermore, IL-1beta-induced IL-8, IL-6, and matrix metalloproteinase-3 protein production was significantly inhibited in DN MKK3/DN MKK6-transfected cells. The constructs had no effect on the respective mediator mRNA levels. These data demonstrate that MKK3 and MKK6 make individual contributions to p38 activation in FLS after cytokine stimulation, but that both must be blocked for maximum inhibition. PMID- 15778395 TI - Targeted overexpression of IL-18 binding protein at the central nervous system overrides flexibility in functional polarization of antigen-specific Th2 cells. AB - The current study shows that functional polarization of Ag-specific CD4(+) Th2 cells entering the CNS during the accelerating phase of experimental autoimmune encephalomyelitis is flexible and dependent on the cytokine milieu there. Thus, targeted cell/gene therapy by Ag-specific T cells overexpressing IL-18 binding protein overrides this flexibility and induces infectious spread of T cell tolerance. Using a congenic system, we demonstrated that at this time, Ag specific Th2 cells accumulate at the CNS but then arrest of IL-4 production. A manipulation of targeted cell/gene delivery was then used to detect whether this function is dependent on the cytokine milieu there. Targeted overexpression of IL 18 binding protein, a natural inhibitor of IL-18, restored the ability of these Ag-specific Th2 cells to produce IL-4 and subsequently induce protective spread of Th2 polarization. These findings not only suggest a novel way of therapy, but also explain why shifting the balance of Ag-specific T cells toward Th2 suppresses ongoing experimental autoimmune encephalomyelitis, whereas a direct transfer of these cells is ineffective. PMID- 15778396 TI - Activation pathways implicate anti-HLA-DP and anti-LFA-1 antibodies as lead candidates for intervention in chronic berylliosis. AB - CD4(+) T cells play a key role in granulomatous inflammation in the lung of patients with chronic beryllium disease. The goal of this study was to characterize activation pathways of beryllium-responsive bronchoalveolar lavage (BAL) CD4(+) T cells from chronic beryllium disease patients to identify possible therapeutic interventional strategies. Our results demonstrate that in the presence of APCs, beryllium induced strong proliferation responses of BAL CD4(+) T cells, production of superoptimal concentrations of secreted proinflammatory cytokines, IFN-gamma, TNF-alpha,and IL-2, and up-regulation of numerous T cell surface markers that would promote T-T Ag presentation. Ab blocking experiments revealed that anti-HLA-DP or anti-LFA-1 Ab strongly reduced proliferation responses and cytokine secretion by BAL CD4(+) T cells. In contrast, anti-HLA-DR or anti-OX40 ligand Ab mainly affected beryllium-induced proliferation responses with little impact on cytokines other than IL-2, thus implying that nonproliferating BAL CD4(+) T cells may still contribute to inflammation. Blockade with CTLA4-Ig had a minimal effect on proliferation and cytokine responses, confirming that activation was independent of B7/CD28 costimulation. These results indicate a prominent role for HLA-DP and LFA-1 in BAL CD4(+) T cell activation and further suggest that specific Abs to these molecules could serve as a possible therapy for chronic beryllium disease. PMID- 15778397 TI - Induction of hyper Th1 cell-type immune responses by dendritic cells lacking the suppressor of cytokine signaling-1 gene. AB - Suppressor of cytokine signaling (SOCS1/JAB) has been shown to play an important role in regulating dendritic cell (DC) function and suppressing inflammatory diseases and systemic autoimmunity. However, role of SOCS1 in DCs for the initiation of Th cell response has not been clarified. Here we demonstrate that SOCS1-deficient DCs induce stronger Th1-type responses both in vitro and in vivo. SOCS1-deficient DCs induced higher IFN-gamma production from naive T cells than wild-type (WT) DCs in vitro. Lymph node T cells also produced a higher amount of IFN-gamma when SOCS1-deficient bone marrow-derived DCs (BMDCs) were transferred in vivo. Moreover, SOCS1(-/-) BMDCs raised more effective anti-tumor immunity than WT BMDCs. Microarray analysis revealed that IFN-inducible genes were highly expressed in SOCS1-deficient DCs without IFN stimulation, suggesting hyper STAT1 activation in SOCS1(-/-) DCs. These phenotypes of SOCS1-deficient DCs were similar to those of CD8alpha(+) DCs, and in the WT spleen, SOCS1 is expressed at higher levels in the Th2-inducing CD4(+) DC subset, relative to the Th1-inducing CD8alpha(+) DC subset. We propose that reduction of the SOCS1 gene expression in DCs leads to CD8alpha(+) DC-like phenotype which promotes Th1-type hyperresponses. PMID- 15778398 TI - HIV-1 transactivator of transcription protein induces mitochondrial hyperpolarization and synaptic stress leading to apoptosis. AB - Despite the efficacy of highly active antiretroviral therapy in reducing viral burden, neurologic disease associated with HIV-1 infection of the CNS has not decreased in prevalence. HIV-1 does not induce disease by direct infection of neurons, although extensive data suggest that intra-CNS viral burden correlates with both the severity of virally induced neurologic disease, and with the generation of neurotoxic metabolites. Many of these molecules are capable of inducing neuronal apoptosis in vitro, but neuronal apoptosis in vivo does not correlate with CNS dysfunction, thus prompting us to investigate cellular and synaptic events occurring before cell death that may contribute to HIV-1 associated neurologic disease. We now report that the HIV-1 regulatory protein transactivator of transcription protein (Tat) increased oxidative stress, ATP levels, and mitochondrial membrane potential in primary rodent cortical neurons. Additionally, a proinflammatory cellular metabolite up-regulated by Tat, platelet activating factor, also induced oxidative stress and mitochondrial hyperpolarization in neurons, suggesting that this type of metabolic dysfunction may occur on a chronic basis during HIV-1 infection of the CNS. Tat-induced mitochondrial hyperpolarization could be blocked with a low dose of the protonophore FCCP, or the mitochondrial KATP channel antagonist, tolbutamide. Importantly, blocking the mitochondrial hyperpolarization attenuated Tat-induced neuronal apoptosis, suggesting that increased mitochondrial membrane potential may be a causal event in precipitating neuronal apoptosis in cell culture. Finally, Tat and platelet-activating factor also increased neuronal vesicular release, which may be related to increased mitochondrial bioenergetics and serve as a biomarker for early damage to neurons. PMID- 15778399 TI - Molecular circuits of resolution: formation and actions of resolvins and protectins. AB - The cellular events underlying the resolution of acute inflammation are not known in molecular terms. To identify anti-inflammatory and proresolving circuits, we investigated the temporal and differential changes in self-resolving murine exudates using mass spectrometry-based proteomics and lipidomics. Key resolution components were defined as resolution indices including Psi(max), the maximal neutrophil numbers that are present during the inflammatory response; T(max), the time when Psi(max) occurs; and the resolution interval (R(i)) from T(max) to T(50) when neutrophil numbers reach half Psi(max). The onset of resolution was at approximately 12 h with proteomic analysis showing both haptoglobin and S100A9 levels were maximal and other exudate proteins were dynamically regulated. Eicosanoids and polyunsaturated fatty acids first appeared within 4 h. Interestingly, the docosahexaenoic acid-derived anti-inflammatory lipid mediator 10,17S-docosatriene was generated during the R(i). Administration of aspirin triggered lipoxin A(4) analog, resolvin E1, or 10,17S-docosatriene each either activated and/or accelerated resolution. For example, aspirin-triggered lipoxin A(4) analog reduced Psi(max), resolvin E1 decreased both Psi(max) and T(max), whereas 10,17S-docosatriene reduced Psi(max), T(max), and shortened R(i). Also, aspirin-triggered lipoxin A(4) analog markedly inhibited proinflammatory cytokines and chemokines at 4 h (20-50% inhibition), whereas resolvin E1 and 10,17S-docosatriene's inhibitory actions were maximal at 12 h (30-80% inhibition). Moreover, aspirin-triggered lipoxin A(4) analog evoked release of the antiphlogistic cytokine TGF-beta. These results characterize the first molecular resolution circuits and their major components activated by specific novel lipid mediators (i.e., resolvin E1 and 10,17S-docosatriene) to promote resolution. PMID- 15778400 TI - Respiratory syncytial virus (RSV) infection induces cyclooxygenase 2: a potential target for RSV therapy. AB - Cyclooxygenases (COXs) are rate-limiting enzymes that initiate the conversion of arachidonic acid to prostanoids. COX-2 is the inducible isoform that is up regulated by proinflammatory agents, initiating many prostanoid-mediated pathological aspects of inflammation. The roles of cyclooxygenases and their products, PGs, have not been evaluated during respiratory syncytial virus (RSV) infection. In this study we demonstrate that COX-2 is induced by RSV infection of human lung alveolar epithelial cells with the concomitant production of PGs. COX 2 induction was dependent on the dose of virus and the time postinfection. PG production was inhibited preferentially by NS-398, a COX-2-specific inhibitor, and indomethacin, a pan-COX inhibitor, but not by SC-560, a COX-1-specific inhibitor. In vivo, COX-2 mRNA expression and protein production were strongly induced in the lungs and cells derived from bronchioalveolar lavage of cotton rats infected with RSV. The pattern of COX-2 expression in vivo in lungs is cyclical, with a final peak on day 5 that correlates with maximal histopathology. Treatment of cotton rats with indomethacin significantly mitigated lung histopathology produced by RSV. The studies described in this study provide the first evidence that COX-2 is a potential therapeutic target in RSV-induced disease. PMID- 15778401 TI - Prenatal blockage of lymphotoxin beta receptor and TNF receptor p55 signaling cascade resulted in the acceleration of tissue genesis for isolated lymphoid follicles in the large intestine. AB - Signaling by lymphotoxin (LT) and TNF is essential for the organogenesis of secondary lymphoid tissues in systemic and mucosal compartments. In this study, we demonstrated that the progeny of mice treated with fusion protein of LTbetaR and IgGFc (LTbetaR-Ig) or LTbetaR-Ig plus TNFR55-Ig (double Ig) showed significantly increased numbers of isolated lymphoid follicles (ILF) in the large intestine. Interestingly, double Ig treatment accelerated the maturation of large intestinal ILF. Three-week-old progeny of double Ig-treated mice showed increased numbers of ILF in the large intestine, but not in the small intestine. Furthermore, alteration of intestinal microflora by feeding of antibiotic water did not affect the increased numbers of ILF in the large intestine of double Ig treated mice. Most interestingly, mice that developed numerous ILF also had increased levels of activation-induced cytidine deaminase expression and numbers of IgA-expressing cells in the lamina propria of the large intestine. Taken together, these results suggest that ILF formation in the large intestine is accelerated by blockage of LTbetaR and TNFR55 signals in utero, and ILF, like colonic patches, might play a role in the induction of IgA response in the large intestine. PMID- 15778402 TI - Protective CD8 T cell immunity triggered by CpG-protein conjugates competes with the efficacy of live vaccines. AB - In contrast to infectious (live) vaccines are those based on subunit Ag that are notoriously poor in eliciting protective CD8 T cell responses, presumably because subunit Ags become insufficiently cross-presented by dendritic cells (DCs) and because the latter need to be activated to acquire competence for cross-priming. In this study, we show that CpG-Ag complexes overcome these limitations. OVA covalently linked to CpG-DNA (CpG-OVA complex), once it is efficiently internalized by DCs via DNA receptor-mediated endocytosis, is translocated to lysosomal-associated membrane protein 1 (LAMP-1)-positive endosomal-lysosomal compartments recently shown to display competence for cross-presentation. In parallel, CpG-OVA complex loaded DCs become activated and acquire characteristics of professional APCs. In vivo, a single s.c. dose of CpG-OVA complex (10 mug of protein) induces primary and secondary clonal expansion/contraction of Ag specific CD8 T cells similar in kinetics to live vaccines; examples including Listeria monocytogenes genetically engineered to produce OVA (LM-OVA) and two viral vector-based OVA vaccines analyzed. Interestingly, CpG-OVA complex induced almost equal percentages of Ag-specific memory CD8 T cells as did infection with LM-OVA. A single dose vaccination with CpG-OVA complex protected mice against lethal doses of LM-OVA. These data underscore that the synergy imparted by CpG OVA complex-mediated combined triggering of innate and specific immunity might be key to initiate CD8 T cell-based immunoprotection by synthetic vaccines based on subunit Ag. PMID- 15778403 TI - In vitro and in vivo characterization of a novel antibody-like single-chain TCR human IgG1 fusion protein. AB - We have constructed a protein composed of a soluble single-chain TCR genetically linked to the constant domain of an IgG1 H chain. The Ag recognition portion of the protein binds to an unmutated peptide derived from human p53 (aa 264-272) presented in the context of HLA-A2.1, whereas the IgG1 H chain provides effector functions. The protein is capable of forming dimers, specifically staining tumor cells and promoting target and effector cell conjugation. The protein also has potent antitumor effects in an in vivo tumor model and can mediate cell killing by Ab-dependent cellular cytotoxicity. Therefore, single-chain TCRs linked to IgG1 H chains behave like Abs but possess the ability to recognize Ags derived from intracellular targets. These fusion proteins represent a novel group of immunotherapeutics that have the potential to expand the range of tumors available for targeted therapies beyond those currently addressed by the conventional Ab-based approach. PMID- 15778404 TI - The peritoneal cavity provides a protective niche for B1 and conventional B lymphocytes during anti-CD20 immunotherapy in mice. AB - Although anti-CD20 immunotherapy effectively treats human lymphoma and autoimmune disease, the in vivo effect of immunotherapy on tissue B cells and their subsets is generally unknown. To address this, anti-mouse CD20 mAbs were used in a mouse model in which the extent and kinetics of tissue B cell depletion could be assessed in vivo. CD20 mAb treatment depleted most mature B cells within 2 days, with 95-98% of B cells in the bone marrow, blood, spleen, lymph nodes, and gut associated lymphoid tissues depleted by day 7, including marginal zone and follicular B cells. The few spleen B cells remaining after CD20 mAb treatment included pre-B, immature, transitional, and some B1 B cells that expressed CD20 at low levels. By contrast, peritoneal cavity B cells expressed normal CD20 densities and were coated with CD20 mAb, but only 30-43% of B1 cells and 43-78% of B2 cells were depleted by day 7. Spleen B cells adoptively transferred into the peritoneal cavity were similarly resistant to mAb-induced depletion, while transferred B cells that had migrated to the spleen were depleted. However, peritoneal B1 and B2 cells were effectively depleted in mAb-treated wild-type and C3-deficient mice by thioglycolate-induced monocyte migration into this otherwise privileged niche. Inflammation-elicited effector cells did not promote peritoneal cavity B cell depletion in FcR-deficient mice treated with CD20 mAb. Thus, the majority of CD20(+) cells and B cell subsets within lymphoid tissues and the peritoneum could be depleted efficiently in vivo through Fc-dependent, but C independent pathways during anti-CD20 immunotherapy. PMID- 15778405 TI - G protein-coupled receptor kinase 2 in multiple sclerosis and experimental autoimmune encephalomyelitis. AB - Many modulators of inflammation, including chemokines, neuropeptides, and neurotransmitters signal via G protein-coupled receptors (GPCR). GPCR kinases (GRK) can phosphorylate agonist-activated GPCR thereby promoting receptor desensitization. Here we describe that in leukocytes from patients with active relapsing-remitting multiple sclerosis (MS) or with secondary progressive MS, GRK2 levels are significantly reduced. Unexpectedly, cells from patients during remission express even lower levels of GRK2. The level of GRK2 in leukocytes of patients after stroke, a neurological disorder with paralysis but without an autoimmune component, was similar to GRK2 levels in cells from healthy individuals. In addition, we demonstrate that the course of recombinant myelin oligodendrocyte glycoprotein (1-125)-induced experimental autoimmune encephalomyelitis (EAE), an animal model for MS, is markedly different in GRK2(+/ ) mice that express 50% of the GRK2 protein in comparison with wild-type mice. Onset of EAE was significantly advanced by 5 days in GRK2(+/-) mice. The earlier onset of EAE was associated with increased early infiltration of the CNS by T cells and macrophages. Although disease scores in the first phase of EAE were similar in both groups, GRK2(+/-) animals did not develop relapses, whereas wild type animals did. The absence of relapses in GRK2(+/-) mice was associated with a marked reduction in inflammatory infiltrates in the CNS. Recombinant myelin oligodendrocyte glycoprotein-induced T cell proliferation and cytokine production were normal in GRK2(+/-) animals. We conclude that down-regulation of GRK2 expression may have important consequences for the onset and progression of MS. PMID- 15778406 TI - Depletion of regulatory T cells in HIV infection is associated with immune activation. AB - Immune activation during chronic HIV infection is a strong clinical predictor of death and may mediate CD4(+) T cell depletion. Regulatory T cells (Tregs) are CD4(+)CD25(bright)CD62L(high) cells that actively down-regulate immune responses. We asked whether loss of Tregs during HIV infection mediates immune activation in a cross-sectional study of 81 HIV-positive Ugandan volunteers. We found that Treg number is strongly correlated with both CD4(+) and CD8(+) T cell activation. In multivariate modeling, this relationship between Treg depletion and CD4(+) T cell activation was stronger than any other clinical factor examined, including viral load and absolute CD4 count. Tregs appear to decline at different rates compared with other CD4(+) T cells, resulting in an increased regulator to helper ratio in many patients with advanced disease. We hypothesize that this skewing may contribute to T cell effector dysfunction. Our findings suggest Tregs are a major contributor to the immune activation observed during chronic HIV infection. PMID- 15778407 TI - Primary human lymphocytes transduced with NY-ESO-1 antigen-specific TCR genes recognize and kill diverse human tumor cell lines. AB - cDNAs encoding TCR alpha- and beta-chains specific for HLA-A2-restricted cancer testis Ag NY-ESO-1 were cloned using a 5'RACE method from RNA isolated from a CTL generated by in vitro stimulation of PBMC with modified NY-ESO-1-specific peptide (p157-165, 9V). Functionality of the cloned TCR was confirmed by RNA electroporation of primary PBL. cDNA for these alpha- and beta-chains were used to construct a murine stem cell virus-based retroviral vector, and high titer packaging cell lines were generated. Gene transfer efficiency in primary T lymphocytes of up to 60% was obtained without selection using a method of precoating retroviral vectors onto culture plates. Both CD4(+) and CD8(+) T cells could be transduced at the same efficiency. High avidity Ag recognition was demonstrated by coculture of transduced lymphocytes with target cells pulsed with low levels of peptide (<20 pM). TCR-transduced CD4 T cells, when cocultured with NY-ESO-1 peptide pulsed T2 cells, could produce IFN-gamma, GM-CSF, IL-4, and IL 10, suggesting CD8-independent, HLA-A2-restricted TCR activation. The transduced lymphocytes could efficiently recognize and kill HLA-A2- and NY-ESO-1-positive melanoma cell lines in a 4-h (51)Cr release assay. Finally, transduced T cells could efficiently recognize NY-ESO-1-positive nonmelanoma tumor cell lines. These results strongly support the idea that redirection of normal T cell specificity by TCR gene transfer can have potential applications in tumor adoptive immunotherapy. PMID- 15778408 TI - Detection of human P2X7 nucleotide receptor polymorphisms by a novel monocyte pore assay predictive of alterations in lipopolysaccharide-induced cytokine production. AB - The nucleotide receptor P2X(7) is expressed by most leukocytes and initiates signaling events that amplify numerous LPS responses. We tested the hypothesis that loss-of-function polymorphisms in the human P2X(7) gene predispose to the production of an anti-inflammatory mediator balance. Accordingly, we developed a novel P2X(7) pore assay in whole blood that magnifies the activity from wild-type alleles and preserves the gene dosage effect for the 1513 C polymorphism (AA, 69 +/- 4; AC, 42 +/- 4; and CC, 6 +/- 1-fold stimulation). Thirty of 200 healthy individuals were identified as having low P2X(7) pore activity. Seven low pore subjects were 1513 CC, 3 and 11 participants had the other known variants 946 GA and 1729 TA respectively; the remaining 9 volunteers likely have novel polymorphisms. Because platelets are a large source of extracellular ATP during inflammation, whole blood was treated ex vivo with Salmonella typhimurium LPS in the absence of exogenous nucleotides. LPS-stimulated whole blood from individuals in the low pore activity group generated reduced plasma levels of TNF-alpha (p = 0.036) and higher amounts of IL-10 (p < 0.001) relative to the high pore controls. This reduction in the TNF-alpha to IL-10 ratio persisted to at least 24 h and is further decreased by cotreatment with 2-methylthio-ATP. The ability of P2X(7) polymorphisms to regulate the LPS-induced TNF-alpha to IL-10 ratio suggests that 15% of healthy adults may exhibit anti-inflammatory mediator responses during major infectious perturbations of the immune system, which can be predicted by P2X(7) pore activity. PMID- 15778409 TI - Does gene therapy become pharmacotherapy? AB - Recent progress in molecular and cellular biology has led to the development of numerous effective cardiovascular drugs. However, there are still a number of diseases for which no known effective therapy exists, such as peripheral arterial disease, ischaemic heart disease, restenosis after angioplasty, and vascular bypass graft occlusion. Currently, gene therapy is emerging as a potential strategy for the treatment of cardiovascular disease despite its limitations. The first human trial in gene therapy for cardiovascular disease was started at 1994 to treat peripheral vascular disease using vascular endothelial growth factor (VEGF). Then, many different potent angiogenic growth factors were tested in clinical trials to treat peripheral arterial disease and ischaemic heart disease. Improvement of clinical symptoms in peripheral arterial disease and ischaemic heart disease has been reported. This review focuses on the future potential of gene therapy for the treatment of cardiovascular disease. In the future, gene therapy might become a real pharmacotherapy to treat cardiovascular disease. PMID- 15778410 TI - Therapeutic angiogenesis and vasculogenesis for tissue regeneration. AB - Therapeutic angiogenesis/vasculogenesis holds promise for the cure of ischaemic disease. The approach postulates the manipulation of spontaneous healing response by supplementation of growth factors or transplantation of vascular progenitor cells. These supplements are intended to foster the formation of arterial collaterals and promote the regeneration of damaged tissues. Angiogenic factors are generally delivered in the form of recombinant proteins or by gene transfer using viral vectors. In addition, new non-viral methods are gaining importance for their safer profile. The association of growth factors with different biological activity might offer distinct advantages in terms of efficacy, yet combined approaches require further optimization. Alternatively, substances with pleiotropic activity might be considered, by virtue of their ability to target multiple mechanisms. For instance, some angiogenic factors not only stimulate the growth of arterioles and capillaries, but also inhibit vascular destabilization triggered by metabolic and oxidative stress. Transplantation of endothelial progenitor cells was recently proposed for the treatment of peripheral and myocardial ischaemia. Progenitor cells can be transplanted either without any preliminary conditioning or after ex vivo genetic manipulation. Delivery of genetically modified progenitor cells eliminates the drawback of immune response against viral vectors and makes feasible repeating the therapeutic procedure in case of injury recurrence. It is envisioned that these new approaches of regenerative medicine will open unprecedented opportunities for the care of life threatening diseases. PMID- 15778411 TI - Cardiovascular pharmacogenomics. AB - There is large interpatient variability in the response to drugs, including cardiovascular drugs. Thus, while some patients achieve the desired therapeutic response from their drug therapy, others do not. There is also a subset of patients who will experience adverse effects, which can range from bothersome to life threatening. Research in recent years has provided compelling evidence that in many cases, genetics contributes importantly to this variable drug response. Thus, pharmacogenomics is a field focused on unravelling the genetic determinants of variable drug response. Examples from the literature of genetic associations with drug efficacy and toxicity are described to provide insight into the field, including the roles of genetic variability in drug-metabolizing enzymes and drug targets. There is also a detailed discussion of the experimental approaches used in cardiovascular pharmacogenomics. Current research is largely focused on a limited candidate gene approach, which allows for description of significant genetic associations with variable response, but often does not explain the genetic basis of variable drug response enough to be useful clinically. As such, there is a move towards genome-wide approaches, and the various technologies available to obtain genomic data are discussed. Cardiovascular pharmacogenomics has the potential for leading to improvements in the use of cardiovascular drug therapy, through selection of the most appropriate drug therapy in an individual based on their genetic information. It will probably be a decade or more before genetic information is widely used in drug therapy decisions, but it seems clear that important findings in the area will continue to expand and the experimental approaches will continue to evolve. PMID- 15778412 TI - Targeting cerebral arteries for gene therapy. AB - After the steady progress towards application of gene therapy to cerebral arterial diseases, several applications, including modification of gene expression in cerebral arteries, are now feasible. There are several possible targets for cerebrovascular gene therapy, and numerous studies have tested gene therapy strategies in animal models of cerebrovascular disorders. However, some major obstacles, especially issues of safety, must be overcome before clinical use in humans. Gene therapy for cerebral arterial diseases is still in its infancy, and many basic and preclinical studies are yet to be done in order to develop effective and safe techniques. PMID- 15778414 TI - The pathogenesis of tuberculosis: the first one hundred (and twenty-three) years. PMID- 15778415 TI - Hemoglobin-mediated, hypoxia-induced vasodilation via nitric oxide: mechanism(s) and physiologic versus pathophysiologic relevance. PMID- 15778417 TI - Origin and uses of primum non nocere--above all, do no harm! AB - The so-called Hippocratic injunction to do no harm has been an axiom central to clinical pharmacology and to the education of medical and graduate students. With the recent reexamination of the nature and magnitude of adverse reactions to drugs, the purposes of this research and review were to discover the origin of this unique Latin expression. It has been reported that the author was neither Hippocrates nor Galen. Searches of writings back to the Middle Ages have uncovered the appearance of the axiom as expressed in English, coupled with its unique Latin, in 1860, with attribution to the English physician, Thomas Sydenham. Commonly used in the late 1800s into the early decades of the 1900s, it was nearly exclusively transmitted orally; it rarely appeared in print in the early 20th century. Its applicability and limitations as a guide to the ethical practice of medicine and pharmacological research are discussed. Despite insufficiencies, it remains a potent reminder that every medical and pharmacological decision carries the potential for harm. PMID- 15778418 TI - Potential utility of data-mining algorithms for early detection of potentially fatal/disabling adverse drug reactions: a retrospective evaluation. AB - The objective of this study was to apply 2 data-mining algorithms to a drug safety database to determine if these methods would have flagged potentially fatal/disabling adverse drug reactions that triggered black box warnings/drug withdrawals in advance of initial identification via "traditional" methods. Relevant drug-event combinations were identified from a journal publication. Data mining algorithms using commonly cited disproportionality thresholds were then applied to the US Food and Drug Administration database. Seventy drug-event combinations were considered sufficiently specific for retrospective data mining. In a minority of instances, potential signals of disproportionate reporting were provided clearly in advance of initial identification via traditional pharmacovigilance methods. Data-mining algorithms have the potential to improve pharmacovigilance screening; however, for the majority of drug-event combinations, there was no substantial benefit of either over traditional methods. They should be considered as potential supplements to, and not substitutes for, traditional pharmacovigilance strategies. More research and experience will be needed to optimize deployment of data-mining algorithms in pharmacovigilance. PMID- 15778419 TI - The safety and effectiveness of newer antiepileptics: a comparative postmarketing cohort study. AB - Clinical trials for the newer antiepileptic drugs (AEDs) have provided inconclusive information to evaluate comparative risk benefit. The authors use data from postmarketing observational cohort studies to compare the failure of treatment with lamotrigine, vigabatrin, and gabapentin in patients with refractory epilepsy. The Drug Safety Research Unit has conducted prescription event monitoring (PEM) studies for lamotrigine, vigabatrin, and gabapentin to monitor their safety when used in primary care. The primary outcome of this study was time to treatment failure in patients who had been prescribed the drug after the start of the PEM study. Patients on gabapentin had reduced time to treatment failure compared to those on the other 2 drugs. The age- and sex-adjusted hazard ratio of failure was 1.53 (95% confidence interval [CI] = 1.38-1.70) for gabapentin compared to lamotrigine and 1.19 (95% CI = 1.10-1.30) for vigabatrin compared to lamotrigine. The observed differences between the 3 study drugs might be confounded by a higher proportion of patients treated with gabapentin having refractory epilepsy, a shorter duration of the gabapentin PEM study, and a lower relative dose of gabapentin (approved at the time of the PEM study). The current study provides information about the routine usage of newer AEDs, which complements evidence from clinical trials regarding the efficacy and safety of these AEDs. Although this study showed differences on times to treatment failure between lamotrigine, vigabatrin, and gabapentin, the results are only useful when considered together with results from other studies seeking to answer the same questions. PMID- 15778420 TI - Review of UCN-01 development: a lesson in the importance of clinical pharmacology. AB - UCN-01 is a protein kinase inhibitor under development as a novel anticancer drug. The initial pharmacologic features in patients were not predicted from preclinical experiments. The distribution volume and the systemic clearance were much lower than those in experimental animals (mice, rats, and dogs), and the elimination half-life was unusually long (>200 hours). The unbound fraction in human plasma was also much smaller than that in dogs, rats and mice, as was the binding of UCN-01 to human alpha-1 acid glycoprotein much stronger than that to human serum albumin or human gamma-globulin. The association constants for alpha 1 acid glycoprotein and human plasma were approximately 8 x 10(8)(mol/L)(-1), indicating extremely high affinity. In this review article, the authors discuss the pharmacologic features of UCN-01 across species and provide a perspective on how this information could be applied prospectively to the future development of this agent. PMID- 15778421 TI - Sequential analysis of tacrolimus dosing in adult lung transplant patients with ABCB1 haplotypes. AB - The genetic polymorphisms in the ABCB1 gene, which encodes for the membrane pump, P-glycoprotein, have been previously demonstrated to have an association with tacrolimus dosing in organ transplant patients. This study associated the haplotype and genotype for ABCB1 G2677T and C3435T variants with a sequential analysis of tacrolimus blood level (ng/mL) per mg/day dosage ([L/D]) administered to 91 adult lung transplant patients at 1, 3, 6, 9, and 12 months after transplantation. Haplotype 22 carriers had a significantly higher tacrolimus [L/D] value in comparison with nonhaplotype 22 carriers (P = .04) only at 1 month after transplant. Sequential analysis demonstrated that ABCB1 genotypes 00 and 01 had low tacrolimus [L/D] values at 1 and 3 months, but these values increased substantially at 6, 9, and 12 months after transplantation. This was not true of the other genotypes with the exception of genotypes 10 and 21, which had small numbers of patients but had consistently low tacrolimus [L/D]. Haplotype analysis also suggested that the homozygous for ABCB1 2677 variant allele had more of an impact on tacrolimus [L/D] in haplotype analysis than that of ABCB1 3435. In conclusion, sequential analysis of tacrolimus [L/D] with haplotypes can explain previous clinical observations of changes in tacrolimus dosage over time but suggests that this effect is limited to individual patient haplotypes. Sequential analysis of drug dosing and haplotypes relationships can provide important information about the induction or inhibition of drug-drug and disease-drug interactions among specific haplotypes. PMID- 15778422 TI - Effect of the MDR1 C3435T variant and P-glycoprotein induction on dicloxacillin pharmacokinetics. AB - This study investigated 2 hypotheses about genotype-phenotype relationships for the efflux transporter, P-glycoprotein: (1) the presence of a synonymous C3435T variant in exon 26 of the MDR1 gene correlates to higher plasma concentrations of a P-glycoprotein substrate, dicloxacillin, and (2) the effects of genotypic differences decrease under conditions of P-glycoprotein induction by rifampin. Eighteen healthy volunteers received two 1-g doses of dicloxacillin, one on the 1st study day and the other on the 11th day of rifampin dosing (600 mg daily). Dicloxacillin and its 5-hydroxymethyl metabolite were analyzed using liquid chromatography/tandem mass spectrometry. Mean dicloxacillin C(max) measurements were 30.5 +/- 13.5, 33.3 +/- 4.7, and 31.1 +/- 12.8 mug/mL in individuals with the CC, CT, and TT genotype at position 3435 in exon 26 of the MDR1 gene. Following rifampin dosing, the mean dicloxacillin C(max) across genotypes decreased from 31.4 +/- 10.8 to 22.9 +/- 7.0 microg/mL (P < .05), whereas the mean oral clearance increased from 235 +/- 82 to 297 +/- 71 mL/min (P < .001), and the mean absorption time increased from 0.71 +/- 0.55 to 1.34 +/- 0.77 h (P < .05). Rifampin treatment increased the formation clearance, C(max), and AUC of the 5-hydroxymethyl metabolite by 135%, 119%, and 59%, respectively. The C3435T variant had no effect on dicloxacillin pharmacokinetics. The data suggested that rifampin induced intestinal P-glycoprotein and increased dicloxacillin metabolism. PMID- 15778423 TI - Studies to investigate the pharmacokinetic interactions between ranolazine and ketoconazole, diltiazem, or simvastatin during combined administration in healthy subjects. AB - The interactions of ranolazine, a new antianginal compound, with inhibitors and substrates of the CYP3A isoenzyme family were studied in 1 open-label and 4 double-blind, randomized, multiple-dose studies. In healthy adult volunteers, the authors sought (1) to determine the steady-state pharmacokinetics, safety, and tolerability of immediate- and sustained-release ranolazine with and without ketoconazole, diltiazem, or simvastatin and (2) to evaluate the effect of ranolazine on the pharmacokinetics of diltiazem, simvastatin, simvastatin metabolites, and HMG-CoA reductase activity. Ketoconazole increased ranolazine plasma concentrations and reduced the CYP3A4-mediated metabolic transformation of ranolazine, confirming that CYP3A4 is the primary metabolic pathway for ranolazine. Diltiazem reduced oral clearance of ranolazine in a dose-dependent manner. Simvastatin did not affect ranolazine pharmacokinetics, although ranolazine increased the AUC and C(max) of simvastatin, simvastatin acid, 2 simvastatin metabolites, and HMG-CoA reductase activity by <2-fold. Administration of ranolazine in combination with diltiazem or simvastatin was safe and well tolerated during the interval studied. PMID- 15778424 TI - The influence of renal function on hydroxyurea pharmacokinetics in adults with sickle cell disease. AB - This was an open-label, nonrandomized, 2-center study conducted to assess the influence of renal function on the pharmacokinetics of hydroxyurea in adults with sickle cell disease (SCD). Seventeen patients were divided into 5 groups: normal renal function (n = 7), mild renal impairment (n = 2), moderate renal impairment (n = 3), severe renal impairment (n = 2), and end-stage renal disease (ESRD, n = 3). Except for patients with ESRD, all the patients received a 15-mg/kg single oral dose of hydroxyurea. Patients with ESRD received a 15-mg/kg oral dose of hydroxyurea on 2 occasions. Blood and urine samples were collected for the assessment of hydroxyurea pharmacokinetics. The results indicate that the systemic exposure increases and the urinary recovery decreases as the degree of renal insufficiency worsens. On the basis of the exposure and the apparent clearance from the current and 2 historical studies, the authors have proposed an initial dosing regimen of hydroxyurea (7.5 mg/kg/day) for SCD patients with CL(cr) <60 mL/min. This dosing strategy is anticipated to provide a safe dose for SCD patients with renal impairment. PMID- 15778425 TI - FTY720 pharmacokinetics in mild to moderate hepatic impairment. AB - The influence of mild and moderate hepatic impairment on FTY720 pharmacokinetics was assessed. The authors enrolled 32 subjects consisting of 8 with mild and 8 with moderate hepatic impairment based on Child-Pugh criteria and 16 demographically matched control subjects. A single 1-mg oral dose of FTY720 was administered under fasting conditions. Blood, plasma, and urine samples were obtained over a 14-day period for measurement of FTY720 and metabolite concentrations and protein binding. Total blood lymphocyte counts and heart rate were serially monitored to assess pharmacologic responses to FTY720. Peak FTY720 blood concentrations were similar across groups. Oral clearance (CL/F) was reduced 10% in mild hepatic impairment (P = .493) and 31% in moderate hepatic impairment (P = .034). There were no significant differences in blood exposure to the hexanoic or butanoic acid metabolites among groups. The effect of FTY720 on blood lymphocytes was similar across groups, with a mean decrease of 44% from the predose value. Like-wise, the effect of FTY720 on supine heart rate was similar across groups, with a mean 13% decrease from the predose rate occurring 2 to 4 hours postdose and recovering within 1 to 2 days. Although hepatic impairment elicited changes in the disposition of FTY720, the magnitude of these changes suggests that the FTY720 dose does not need to be adjusted in mild or moderate hepatic-impaired patients. PMID- 15778426 TI - Safety, tolerability, and pharmacokinetics of a capsule formulation of DRF-1042, a novel camptothecin analog, in refractory cancer patients in a bridging phase I study. AB - The purpose of this bridging phase I study was to characterize the toxicity, pharmacokinetics, and antitumor effects of a capsule formulation of DRF-1042, a novel camptothecin analog, in refractory solid tumor patients. DRF-1042 was given daily for 5 consecutive days for 2 weeks, repeated every 3 weeks at 81 mg/m(2). Adverse events were monitored following NCI-CTC. Blood samples were processed for bioanalysis using a validated high-performance liquid chromatography method. The pharmacokinetics of lactone and total (lactone + carboxylate) forms was determined on days 1 and 12 using a noncompartmental pharmacokinetic method. Pharmacokinetic data with the capsule formulation were compared with previously reported pharmacokinetic parameters with a suspension formulation. Efficacy was evaluated by applying World Health Organization criteria. Six patients received 10 courses of therapy. Thrombocytopenia and diarrhea were dose-limiting toxicities. The upper limit of the area under the curve of DRF-1042 (lactone and total) with the capsule formulation was higher than a suspension formulation at a similar dose on day 1 (lactone: capsule = 8.53 microMxh, suspension = 5.33 microMxh; total: capsule = 393 microMxh, suspension = 176 microMxh) and day 12 (lactone: capsule = 22.1 microMxh, suspension = 6.1 microMxh; total: capsule = 1302 microMxh, suspension = 309 microMxh). The upper limit of the area under the curve of DRF-1042 (lactone and total) was higher under fed conditions (lactone = 15.9 microMxh, total = 605 microMxh) relative to fasted conditions (lactone = 8.53 microMxh, total = 393 microMxh) on day 1. One patient experienced stable disease. The toxicity and pharmacokinetics of the capsule correlated well with the suspension. The recommended phase II dose is 81 mg/m(2). PMID- 15778427 TI - Successful projection of the time course of drug concentration in plasma during a 1-year period from electronically compiled dosing-time data used as input to individually parameterized pharmacokinetic models. AB - Pharmacokinetic studies rely on blood sampling at times relative to predefined dosing intervals. Intensive sampling is often done under direct observation of dose taking, which, though costly, virtually eliminates uncertainty about actual dosing times. In contrast, the sparse sampling done in population pharmacokinetic studies relies on patient-reported times of dosing, the accuracy of which the authors sought to assess by adding electronic monitoring to the usual patient reporting of dosing times. The study involved 35 antiretroviral-naive, human immunodeficency virus-infected patients and was designed to assess the safety, tolerability, pharmacokinetics, and antiviral activity of prescribed lopinavir/ritonavir (800/200 mg qd or 400/100 mg bid), stavudine, and lamivudine. The present research reports the pharmacokinetic analysis that results from taking into account the patients' actual dosing histories. Intensive sampling for plasma lopinavir concentrations was done at week 3, and 4 additional predose (trough) concentrations were measured during the next 12 months. Convergence was achieved by fitting a simple 1-compartment pharmacokinetic model, with first order absorption and elimination, to the sparse sampling data, using electronic monitoring-reported times. In contrast, convergence was not achieved using the simple model when steady state was assumed, and the times for the last qd dose or the last 2 bid doses, as reported by the patient, were used as model input. Estimated individual pharmacokinetic parameters were then combined with electronic dosing histories to project each patient's internal drug exposure over long periods of time. This strategy may provide a basis for greatly increasing the informational yield and utility of conventional therapeutic drug monitoring. PMID- 15778428 TI - Population pharmacokinetics of efalizumab (humanized monoclonal anti-CD11a antibody) following long-term subcutaneous weekly dosing in psoriasis subjects. AB - The population pharmacokinetics of efalizumab was characterized in patients with moderate to severe plaque psoriasis. The study included 1088 subjects who received 1 or 2 mg/kg/wk subcutaneous efalizumab for 12 weeks from a phase I (64 subjects) and 3 phase III studies with day 42 and/or day 84 trough levels (1024 patients). Due to the limitation of the data, a 1-compartment model with first order absorption and elimination was used to fit the data. The population means for V/F, Ka, and CL/F were 9.13 L, 0.191 day(-1), and 1.29 L/d, respectively, for a typical subject receiving a 1-mg/kg dose. Interindividual variability in CL/F was 48.2%. Body weight has the largest influence on CL/F. Other covariates (obesity, baseline lymphocyte counts, Psoriasis Area and Severity Index score, and age) had only modest effects. Subjects in the 2-mg/kg dose group had a 24.0% lower CL/F, consistent with nonlinear pharmacokinetics of efalizumab. The results of this analysis support the current body weight-adjusted dosing strategy. PMID- 15778429 TI - A pilot study of netilmicin pharmacokinetics during continuous venovenous hemodiafiltration. PMID- 15778430 TI - Enhanced ghrelin secretion in rats with cysteamine-induced duodenal ulcers. AB - Ghrelin, produced and secreted by the A-like cells of the stomach, stimulates growth hormone secretion, gastric motility, and food intake. Cysteamine inhibits the release of somatostatin and induces the formation of duodenal ulcers in rats. The present study was conducted to investigate the dynamics of ghrelin secretion in rats treated with cysteamine. Male Wistar rats (7 wk old) were administered three doses of cysteamine (400 mg/kg) orally; at 50 h after the first dose, duodenal ulcers were induced, and the plasma level of somatostatin and gastric density of somatostatin-immunoreactive cells were significantly reduced. The plasma total and active ghrelin levels were significantly higher in the cysteamine-treated rats than in the control rats, whereas the gastric ghrelin levels, number of gastric ghrelin-immunoreactive cells, and preproghrelin mRNA expression levels were significantly lower. Even at the time points of 2 and 10 h after the first dose of cysteamine, at which time no significant ulcer formation or antral neutrophil accumulation was yet noted, a significant increase in the plasma ghrelin level and decrease in the gastric ghrelin level were observed. Furthermore, although lansoprazole treatment attenuated the duodenal ulceration induced by cysteamine, the increase in the plasma level of ghrelin could still be demonstrated. Because an inverse correlation was found between the plasma ghrelin and somatostatin levels, the inhibition of somatostatin secretion may be associated with the increased ghrelin secretion. In conclusion, an increase in the plasma ghrelin level precedes the formation of duodenal ulcers in rats treated with cysteamine. PMID- 15778431 TI - TGF-beta1 gene transfer to the mouse colon leads to intestinal fibrosis. AB - Crohn's disease (CD) is a chronic, relapsing inflammatory bowel disease, characterized by transmural inflammation. In CD, the recurrent inflammatory injury and tissue repair that occurs in the intestine can progress uncontrollably, leading to the proliferation of mesenchymal cells as well as fibrosis, characterized by excessive extracellular matrix deposition. These processes thicken the bowel wall, reducing flexibility, and often culminate in obstructive strictures. Because no effective measures are currently available to specifically treat or prevent intestinal stricturing, we sought to gain a better understanding of its pathogenesis by developing a mouse model of intestinal fibrosis. Because transforming growth factor (TGF)-beta1 can mediate both fibrosis and mesenchymal cell proliferation; we studied the effects of delivering adenoviral vectors encoding spontaneously active TGF-beta1 into the colons of mice. We first demonstrated that enema delivery of marker adenoviral vectors led to the transfection of the colonic epithelium and transient transgene expression. Histologically, control vectors caused an acute inflammatory response, involving the recruitment of neutrophils and mononuclear cells into the colonic lamina propria; however, infection caused little if any fibrosis. In contrast, the TGF beta1 vector caused a more severe and prolonged inflammatory response as well as localized collagen deposition, leading to severe and progressive fibrosis. This was accompanied by the emergence of cells with a myofibroblast phenotype. Ultimately the fibrosis resulted in many of the TGF-beta1-transfected mice developing profound colonic obstruction. Through adenoviral gene transfer technology, we describe a novel mouse model of colitis and implicate TGF-beta1 in the pathogenesis of obstructive intestinal fibrosis. PMID- 15778433 TI - The effect of the 2-amino group of 7,8-dihydro-8-oxo-2'-deoxyguanosine on translesion synthesis and duplex stability. AB - Replication of DNA containing 7,8-dihydro-8-oxo-2'-deoxyguanosine (OxodG) gives rise to G --> T transversions. The syn-isomer of the lesion directs misincorporation of 2'-deoxyadenosine (dA) opposite it. We investigated the role of the 2-amino substituent on duplex thermal stability and in replication using 7,8-dihydro-8-oxo-2'-deoxyinosine (OxodI). Oligonucleotides containing OxodI at defined sites were chemically synthesized via solid phase synthesis. Translesion incorporation opposite OxodI was compared with 7,8-dihydro-8-oxo-2' deoxyguanosine (OxodG), 2'-deoxyinosine (dI) and 2'-deoxyguanosine (dG) in otherwise identical templates. The Klenow exo- fragment of Escherichia coli DNA polymerase I incorporated 2'-deoxyadenosine (dA) six times more frequently than 2'-deoxycytidine (dC) opposite OxodI. Preferential translesion incorporation of dA was unique to OxodI. UV-melting experiments revealed that DNA containing OxodI opposite dA is more stable than when the modified nucleotide is opposed by dC. These data suggest that while duplex DNA accommodates the 2-amino group in syn OxodG, this substituent is thermally destabilizing and does not provide a kinetic inducement for replication by Klenow exo-. PMID- 15778434 TI - Conversion of DNA methyltransferases into azidonucleosidyl transferases via synthetic cofactors. AB - Aziridine-based cofactor mimics have been synthesized and are shown to undergo methyltransferase-dependent DNA alkylation. Notably, each cofactor mimic possesses an azide functionality, to which can be attached an assortment of unnatural groups following methyltransferase-dependent DNA delivery. DNA duplexes modified with these cofactor mimics are capable of undergoing the Staudinger ligation with phosphines tethered to biological functionalities following enzymatic modification. This methodology provides a new tool by which to selectively modify DNA in a methyltransferase-dependent way. The conversion of biological methyltransferases into azidonucleosidyl transferases demonstrated here also holds tremendous promise as a means of identifying, as yet, unknown substrates of methylation. PMID- 15778432 TI - Two modes of microsatellite instability in human cancer: differential connection of defective DNA mismatch repair to dinucleotide repeat instability. AB - Microsatellite instability (MSI) is associated with defective DNA mismatch repair in various human malignancies. Using a unique fluorescent technique, we have observed two distinct modes of dinucleotide microsatellite alterations in human colorectal cancer. Type A alterations are defined as length changes of < or =6 bp. Type B changes are more drastic and involve modifications of > or =8 bp. We show here that defective mismatch repair is necessary and sufficient for Type A changes. These changes were observed in cell lines and in tumours from mismatch repair gene-knockout mice. No Type B instability was seen in these cells or tumours. In a panel of human colorectal tumours, both Type A MSI and Type B instability were observed. Both types of MSI were associated with hMSH2 or hMLH1 mismatch repair gene alterations. Intriguingly, p53 mutations, which are generally regarded as uncommon in human tumours of the MSI+ phenotype, were frequently associated with Type A instability, whereas none was found in tumours with Type B instability, reflecting the prevailing viewpoint. Inspection of published data reveals that the microsatellite instability that has been observed in various malignancies, including those associated with Hereditary Non-Polyposis Colorectal Cancer (HNPCC), is predominantly Type B. Our findings indicate that Type B instability is not a simple reflection of a repair defect. We suggest that there are at least two qualitatively distinct modes of dinucleotide MSI in human colorectal cancer, and that different molecular mechanisms may underlie these modes of MSI. The relationship between MSI and defective mismatch repair may be more complex than hitherto suspected. PMID- 15778435 TI - Kinetic properties of DM-nitrophen binding to calcium and magnesium. AB - Caged-Ca(2+) compounds such as nitrophenyl-EGTA (NP-EGTA) and DM-nitrophen (DMn) are extremely useful in biological research, but their use in live cells is hampered by cytoplasmic [Mg(2+)]. We determined the properties of Ca(2+) release from NP-EGTA and DMn by using Oregon green BAPTA-5N to measure changes in [Ca(2+)] after ultraviolet flash photolysis in vitro, with or without Mg(2+) present. A large fraction (65%) of NP-EGTA, which has a negligible Mg(2+) affinity, uncages with a time constant of 10.3 ms, resulting in relatively slow increases in [Ca(2+)]. Uncaging of DMn is considerably faster, but DMn has a significant affinity for Mg(2+) to complicate the uncaging process. With experimentally determined values for the Ca(2+) and Mg(2+) binding/unbinding rates of DMn and NP-EGTA, we built a mathematical model to assess the utility of NP-EGTA and DMn in rapid Ca(2+)-uncaging experiments in the presence of Mg(2+). We discuss the advantages and disadvantages of using each compound under different conditions. To determine the kinetics of Ca(2+) binding to biologically relevant Ca(2+) buffers, such as Ca(2+)-binding proteins, the use of DMn is preferable even in the presence of Mg(2+). PMID- 15778436 TI - Lipid bilayer perturbations around a transmembrane nanotube: a coarse grain molecular dynamics study. AB - The perturbations induced in a lipid bilayer by the presence of a transmembrane nanotube are investigated using coarse grained molecular dynamics. Meniscus formation by the lipids and tilting of the nanotube occur in response to hydrophobic mismatch, although these two effects do not compensate completely for the total mismatch. The lipid head-to-tail vector field is examined and shows strong ordering in the membrane plane regardless of the nanotube length. Molecular layering at the lipid-nanotube interface is reported. This study extends previous theoretical approaches to a more realistic setting. PMID- 15778437 TI - Nanosecond dynamics of a mimicked membrane-water interface observed by time resolved stokes shift of LAURDAN. AB - We studied the dipolar relaxation of the surfactant-water interface in reverse micelles of AOT-water in isooctane in the nanosecond and subnanosecond time ranges by incorporating the amphipathic solvatochromic fluorescent probes LAURDAN and TOE. A negative component was observed in the fluorescence decays in the red edge of the emission spectrum-the signature of an excited state reaction-with LAURDAN but not for TOE. The deconvolution of the transient reconstructed spectra of LAURDAN based on a model constructed by adding together three log-normal Gaussian equations made it possible to separate the specific dynamic solvent response from the intramolecular excited state reactions of the probe. The deconvoluted spectrum of lowest energy displayed the largest Stokes shift. This spectral shift was described by unimodal kinetics on the nanosecond timescale, whereas the relaxation kinetics of water-soluble probes have been reported to be biphasic (on the subnanosecond and nanosecond timescales) due to the heterogeneous distribution of these probes in the water pool. Most of this spectral shift probably resulted from water relaxation as it was highly sensitive to the water to surfactant molar ratio (w(0)) (60-65 nm at w(0) = 20-30). A small part of this spectral shift (9 nm at w(0) = 0) probably resulted from dipolar interaction with the AOT polar headgroup. The measured relaxation time values were in the range of the rotational motion of the AOT polar headgroup region as assessed by LAURDAN and TOE fluorescence anisotropy decays. PMID- 15778438 TI - Na+ shows a markedly higher potential than K+ in DNA compaction in a crowded environment. AB - Whereas many physicochemical investigations have shown that among monovalent cations Na(+) ion possesses minimal potential for DNA binding, biological assays have shown that Na(+) ion (in contrast to K(+) ion) plays a primary role in chromatin compaction and related processes. It is difficult to explain this inverse relationship between the compaction potentials of Na(+) and K(+) and their binding abilities. In this study we sought to resolve this contradiction and emphasize the phenomenological distinction between DNA compaction and DNA binding processes in the case of DNA compaction by monocations. Using polyethylene glycol solutions as a model of a crowded cell environment, we studied DNA compaction by alkali metal salts LiCl, NaCl, KCl, RbCl, and CsCl, and found that all of these monocations promote DNA compaction. Among these monovalent cations Na(+) produces the greatest compaction and the ratio of K(+) cand Na(+) oncentrations for DNA compaction is approximately 1.5-2. A comparative analysis of recent experimental results indicates that a higher binding activity of monocation generally corresponds to a low compaction potential of the corresponding monovalent ion. This inverse relation is explained as a result of partial dehydration of monocations in the compact state. PMID- 15778439 TI - Braiding DNA: experiments, simulations, and models. AB - DNA encounters topological problems in vivo because of its extended double helical structure. As a consequence, the semiconservative mechanism of DNA replication leads to the formation of DNA braids or catenanes, which have to be removed for the completion of cell division. To get a better understanding of these structures, we have studied the elastic behavior of two braided nicked DNA molecules using a magnetic trap apparatus. The experimental data let us identify and characterize three regimes of braiding: a slightly twisted regime before the formation of the first crossing, followed by genuine braids which, at large braiding number, buckle to form plectonemes. Two different approaches support and quantify this characterization of the data. First, Monte Carlo (MC) simulations of braided DNAs yield a full description of the molecules' behavior and their buckling transition. Second, modeling the braids as a twisted swing provides a good approximation of the elastic response of the molecules as they are intertwined. Comparisons of the experiments and the MC simulations with this analytical model allow for a measurement of the diameter of the braids and its dependence upon entropic and electrostatic repulsive interactions. The MC simulations allow for an estimate of the effective torsional constant of the braids (at a stretching force F = 2 pN): C(b) approximately 48 nm (as compared with C approximately 100 nm for a single unnicked DNA). Finally, at low salt concentrations and for sufficiently large number of braids, the diameter of the braided molecules is observed to collapse to that of double-stranded DNA. We suggest that this collapse is due to the partial melting and fraying of the two nicked molecules and the subsequent right- or left-handed intertwining of the stretched single strands. PMID- 15778440 TI - Dynamics of myosin-driven skeletal muscle contraction: I. Steady-state force generation. AB - Skeletal muscle contraction is a canonical example of motor-driven force generation. Despite the long history of research in this topic, a mechanistic explanation of the collective myosin force generation is lacking. We present a theoretical model of muscle contraction based on the conformational movements of individual myosins and experimentally measured chemical rate constants. Detailed mechanics of the myosin motor and the geometry of the sarcomere are taken into account. Two possible scenarios of force generation are examined. We find only one of the scenarios can give rise to a plausible contraction mechanism. We propose that the synchrony in muscle contraction is due to a force-dependent ADP release step. Computational results of a half sarcomere with 150 myosin heads can explain the experimentally measured force-velocity relationship and efficiency data. We predict that the number of working myosin motors increases as the load force is increased, thus showing synchrony among myosin motors during muscle contraction. We also find that titin molecules anchoring the thick filament are passive force generators in assisting muscle contraction. PMID- 15778441 TI - Membrane fluidity is a key modulator of membrane binding, insertion, and activity of 5-lipoxygenase. AB - Mammalian 5-lipoxygenase (5-LO) catalyzes conversion of arachidonic acid to leukotrienes, potent mediators of inflammation and allergy. Upon cell stimulation, 5-LO selectively binds to nuclear membranes and becomes activated, yet the mechanism of recruitment of 5-LO to nuclear membranes and the mode of 5 LO-membrane interactions are poorly understood. Here we show that membrane fluidity is an important determinant of membrane binding strength of 5-LO, penetration into the membrane hydrophobic core, and activity of the enzyme. The membrane binding strength and activity of 5-LO increase with the degree of lipid acyl chain cis-unsaturation and reach a plateau with 1-palmitoyl-2-arachidonolyl sn-glycero-3-phosphocholine (PAPC). A fraction of tryptophans of 5-LO penetrate into the hydrocarbon region of fluid PAPC membranes, but not into solid 1,2 dipalmitoyl-sn-glycero-3-phosphocholine membranes. Our data lead to a novel concept of membrane binding and activation of 5-LO, suggesting that arachidonic acid-containing lipids, which are present in nuclear membranes at higher fractions than in other cellular membranes, may facilitate preferential membrane binding and insertion of 5-LO through increased membrane fluidity and may thereby modulate the activity of the enzyme. The data presented in this article and earlier data allow construction of a model for membrane-bound 5-LO, including the angular orientation and membrane insertion of the protein. PMID- 15778442 TI - Lateral ligand-receptor interactions on membranes probed by simultaneous fluorescence-interference detection. AB - We describe an experimental approach for studying ligand-receptor interactions in the plane of the membrane. The extracellular domains of the type I interferon receptor subunits ifnar1-EC and ifnar2-EC were tethered in an oriented fashion onto solid-supported, fluid lipid bilayers, thus mimicking membrane anchoring and lateral diffusion of the receptor. Ligand-induced receptor assembling was investigated by simultaneous total internal reflection fluorescence spectroscopy and reflectance interferometry (RIf). Based on a rigorous characterization of the interactions of fluorescence-labeled IFNalpha2 with each of the receptor subunits, the dynamics of the ternary complex formation on the fluid lipid bilayer was addressed in further detail making use of the features of the simultaneous detection. All these measurements supported the formation of a ternary complex in two steps, i.e., association of the ligand to ifnar2-EC and subsequent recruitment of ifnar1-EC on the surface of the membrane. Based on the ability to control and quantify the receptor surface concentrations, equilibrium, and rate constants of the interaction in the plane of the membrane were determined by monitoring ligand dissociation at different receptor surface concentrations. Using mutants of IFNalpha2 binding to ifnar2-EC with different association rate constants, the key role of the association rate constants for the assembling mechanism was demonstrated. PMID- 15778443 TI - Gapped DNA and cyclization of short DNA fragments. AB - We use the cyclization of small DNA molecules, approximately 200 bp in length, to study conformational properties of DNA fragments with single-stranded gaps. The approach is extremely sensitive to DNA conformational properties and, being complemented by computations, allows a very accurate determination of the fragment's conformational parameters. Sequence-specific nicking endonucleases are used to create the 4-nt-long gap. We determined the bending rigidity of the single-stranded region in the gapped DNA. We found that the gap of 4 nt in length makes all torsional orientations of DNA ends equally probable. Our results also show that the gap has isotropic bending rigidity. This makes it very attractive to use gapped DNA in the cyclization experiments to determine DNA conformational properties, since the gap eliminates oscillations of the cyclization efficiency with the DNA length. As a result, the number of measurements is greatly reduced in the approach, and the analysis of the data is greatly simplified. We have verified our approach on DNA fragments containing well-characterized intrinsic bends caused by A-tracts. The obtained experimental results and theoretical analysis demonstrate that gapped-DNA cyclization is an exceedingly sensitive and accurate approach for the determination of DNA bending. PMID- 15778444 TI - Topography and mechanical properties of single molecules of type I collagen using atomic force microscopy. AB - Although the mechanical behavior of tendon and bone has been studied for decades, there is still relatively little understanding of the molecular basis for their specific properties. Thus, despite consisting structurally of the same type I collagen, bones and tendons have evolved to fulfill quite different functions in living organisms. In an attempt to understand the links between the mechanical properties of these collageneous structures at the macro- and nanoscale, we studied trimeric type I tropocollagen molecules by atomic force microscopy, both topologically and by force spectroscopy. High-resolution imaging demonstrated a mean (+/- SD) contour length of (287 +/- 35) nm and height of (0.21 +/- 0.03) nm. Submolecular features, namely the coil-pitch of the molecule, were also observed, appearing as a repeat pattern along the length of the molecule, with a length of approximately 8 nm that is comparable to the theoretical value. Using force spectroscopy, we established the stretching pattern of the molecule, where both the mechanical response of the molecule and pull-off peak are convoluted in a single feature. By interpreting this response with a wormlike chain model, we extracted the value of the effective contour length of the molecule at (202 +/- 5) nm. This value was smaller than that given by direct measurement, suggesting that the entire molecule was not being stretched during the force measurements; this is likely to be related to the absence of covalent binding between probe, sample, and substrate in our experimental procedure. PMID- 15778446 TI - The kinetics of phase separation in asymmetric membranes. AB - Phase separation in a model asymmetric membrane is studied using Monte Carlo techniques. The membrane comprises two species of particles, which mimic different lipids in lipid bilayers and separately possess either zero or non-zero spontaneous curvatures. We study the influence of phase separation on membrane shape and the influence of the coupling of composition and height dynamics on phase separation and domain growth, via both the degree of shape asymmetry and relative kinetic coefficients for height relaxation. PMID- 15778445 TI - Oxidation and reduction control of the inactivation gating of Torpedo ClC-0 chloride channels. AB - Oxidation and reduction (redox) are known to modulate the function of a variety of ion channels. Here, we report a redox regulation of the function of ClC-0, a chloride (Cl(-)) channel from the Torpedo electric organ. The study was motivated by the occasional observation of oocytes with hyperpolarization-activated Cl(-) current when these oocytes expressed ClC-0. We find that these atypical recording traces can be turned into typical ClC-0 current by incubating the oocyte in millimolar concentrations of reducing agents, suggesting that the channel function is regulated by oxidation and reduction. The redox control apparently results from an effect of oxidation on the slow (inactivation) gating: oxidation renders it more difficult for the channel to recover from the inactivated states. Introducing the point mutation C212S in ClC-0 suppresses the inactivation state, and this inactivation-suppressed mutant is no longer sensitive to the inhibition by oxidizing reagents. However, C212 is probably not the target for the redox reaction because the regulation of the inactivation gating by oxidation is still present in a pore mutant (K165C/K165 heterodimer) in which the C212S mutation is present. Taking advantage of the K165C/K165 heterodimer, we further explore the oxidation effect in ClC-0 by methane thiosulfonate (MTS) modifications. We found that trimethylethylammonium MTS modification of the introduced cysteine can induce current in the K165C/K165 heterodimer, an effect attributed to the recovery of the channel from the inactivation state. The current induction by MTS reagents is subjected to redox controls, and thus the extent of this current induction can serve as an indicator to report the oxidation state of the channel. These results together suggest that the inactivation gating of ClC-0 is affected by redox regulation. The finding also provides a convenient method to "cure" those atypical recording traces of ClC-0 expressed in Xenopus oocytes. PMID- 15778447 TI - Does the chromophore's ring move after photoexcitation of rhodopsin? AB - By comparing the shift of the absorption maxima when a visual pigment is converted to its lumirhodopsin photointermediate for two classes of pigments, we can infer whether or not the pigment's beta-ionone ring has left its binding site. We compare this shift for the long-wavelength sensitive visual pigment of chicken iodopsin (lambdamax = 571 nm), which has polar residues in the ring binding site that interact with the ring, with that for three pigments, which do not. We conclude that by the time the Lumi product of the pigment is formed, the ring has moved away from the ring binding site. PMID- 15778448 TI - Determination of the orientation of T4 lysozyme vectorially bound to a planar supported lipid bilayer using site-directed spin labeling. AB - Site-directed spin labeling is used to investigate the structure of adsorbed T4 lysozyme (T4L). A monolayer of T4L is prepared by tethering the protein selectively via a His-tag to the chelating headgroups (NTA Ni) of a planar quartz supported lipid bilayer. This results in a vectorially oriented ensemble of proteins on the surface, which gives rise to angular-dependent electron paramagnetic resonance spectra. Similar measurements of spin-labeled lipid bilayers were used to characterize the structure and dynamics of the supports. Electron paramagnetic resonance line shape was analyzed using the stochastic Liouville equation approach developed by Freed and co-workers. The simulations reveal a conservation of the secondary and tertiary structure of T4L upon adsorption although slight conformational changes in the presence of the surface can be detected by probing tertiary contact sites. The orientation of the entire protein was deduced on the basis of an anisotropic motional model for the spin labeled side chain. In addition, a polar order but azimuthal disorder of the molecules was assumed to fit the data. These results demonstrate the utility of site-directed spin labeling in combination with spectral simulation to study not only the secondary and tertiary structure of adsorbed proteins in monolayer coverage but also their orientation with respect to the surface. PMID- 15778449 TI - The single chlorophyll a molecule in the cytochrome b6f complex: unusual optical properties protect the complex against singlet oxygen. AB - The cytochrome b(6)f complex of oxygenic photosynthesis mediates electron transfer between the reaction centers of photosystems I and II and facilitates coupled proton translocation across the membrane. High-resolution x-ray crystallographic structures (Kurisu et al., 2003; Stroebel et al., 2003) of the cytochrome b(6)f complex unambiguously show that a Chl a molecule is an intrinsic component of the cytochrome b(6)f complex. Although the functional role of this Chl a is presently unclear (Kuhlbrandt, 2003), an excited Chl a molecule is known to produce toxic singlet oxygen as the result of energy transfer from the excited triplet state of the Chl a to oxygen molecules. To prevent singlet oxygen formation in light-harvesting complexes, a carotenoid is typically positioned within approximately 4 A of the Chl a molecule, effectively quenching the triplet excited state of the Chl a. However, in the cytochrome b(6)f complex, the beta carotene is too far (> or =14 Angstroms) from the Chl a for effective quenching of the Chl a triplet excited state. In this study, we propose that in this complex, the protection is at least partly realized through special arrangement of the local protein structure, which shortens the singlet excited state lifetime of the Chl a by a factor of 20-25 and thus significantly reduces the formation of the Chl a triplet state. Based on optical ultrafast absorption difference experiments and structure-based calculations, it is proposed that the Chl a singlet excited state lifetime is shortened due to electron exchange transfer with the nearby tyrosine residue. To our knowledge, this kind of protection mechanism against singlet oxygen has not yet been reported for any other chlorophyll-containing protein complex. It is also reported that the Chl a molecule in the cytochrome b(6)f complex does not change orientation in its excited state. PMID- 15778451 TI - Opioid receptor homo- and heterodimerization in living cells by quantitative bioluminescence resonance energy transfer. AB - Opioid receptors have been shown to dimerize or oligomerize among themselves and each other, affecting their functional properties. This study used bioluminescence resonance energy transfer (BRET) between the mu, delta, and kappa opioid receptors to study opioid receptor aggregation in transfected human embryonic kidney 293 cells. Titration of receptor levels indicated that all three opioid receptors have a similar affinity to form homo- or hetero-oligomers in combination with any other opioid receptor type. In contrast, none of the opioid receptors formed detectable oligomers with the muscarinic M2 receptor, indicating that interactions among opioid receptors are selective. The formation of opioid receptor dimers, rather than higher order oligomers, is supported by binding kinetics in competition experiments between labeled and unlabeled receptors. Opioid receptor dimerization occurred at physiological temperatures upon receptor biosynthesis, before trafficking to the plasma membrane. Moreover, using BRET, coimmunoprecipitation, receptor binding, and G protein coupling, we demonstrate for the first time functional mu opioid receptor-kappa opioid receptor heterodimerization. These combined results demonstrate that opioid receptors can undergo homo- and heterodimerization, a process with potential implications for opioid physiology and pharmacology. PMID- 15778450 TI - Peptides derived from apoptotic Bax and Bid reproduce the poration activity of the parent full-length proteins. AB - Bax and Bid are proapoptotic proteins of the Bcl-2 family that regulate the release of apoptogenic factors from mitochondria. Although they localize constitutively in the cytoplasm, their apoptotic function is exerted at the mitochondrial outer membrane, and is related to their ability to form transbilayer pores. Here we report the poration activity of fragments from these two proteins, containing the first alpha-helix of a colicinlike hydrophobic hairpin (alpha-helix 5 of Bax and alpha-helix 6 of Bid). Both peptides readily bind to synthetic lipid vesicles, where they adopt predominantly alpha-helical structures and induce the release of entrapped calcein. In planar lipid membranes they form ion conducting channels, which in the case of the Bax-derived peptide are characterized by a two-stage pattern, a large conductivity and lipid-charge dependent ionic selectivity. These features, together with the influence of intrinsic lipid curvature on the poration activity and the existence of two helical stretches of different orientations for the membrane-bound peptide, suggest that it forms mixed lipidic/peptidic pores of toroidal structure. In contrast, the assayed Bid fragment shows a markedly different behavior, characterized by the formation of discrete, steplike channels in planar lipid bilayers, as expected for a peptidic pore lined by a bundle of helices. PMID- 15778452 TI - Tristetraprolin regulates the expression of the human inducible nitric-oxide synthase gene. AB - The expression of human inducible NO synthase (iNOS) is regulated both by transcriptional and post-transcriptional mechanisms. Stabilization of mRNAs often depends on activation of p38 mitogen-activated protein kinase (p38 MAPK). In human DLD-1 cells, inhibition of p38 MAPK by the compound 4-(4-fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB203580) or by overexpression of a dominant-negative p38 MAPKalpha protein resulted in a reduction of human iNOS mRNA and protein expression, whereas human iNOS promoter activity was not affected. An important RNA binding protein regulated by the p38 MAPK pathway and involved in the regulation of the stability of several mRNAs is tristetraprolin. RNase protection, quantitative real-time polymerase chain reaction, and Western blot experiments showed that cytokines used to induce iNOS expression in DLD-1 cells also enhanced tristetraprolin expression. SB203580 incubation reduced cytokine-mediated enhancement of tristetraprolin expression. Overexpression or down-regulation of tristetraprolin in stably transfected DLD-1- or A549/8 cells consistently resulted in enhanced or reduced iNOS expression by modulating iNOS mRNA stability. In UV cross-linking experiments, recombinant tristetraprolin did not interact with the human iNOS mRNA. However, coimmunoprecipitation experiments showed interaction of tristetraprolin with the KH-type splicing regulatory protein (KSRP), which is known to recruit mRNAs containing AU-rich elements to the exosome for degradation. This tristetraprolin-KSRP interaction was enhanced by cytokines and reduced by SB203580 treatment. We conclude that tristetraprolin positively regulates human iNOS expression by enhancing the stability of human iNOS mRNA. Because tristetraprolin does not directly bind to the human iNOS mRNA but interacts with KSRP, tristetraprolin is likely to stabilize iNOS mRNA by capturing the KSRP-exosome complex. PMID- 15778453 TI - Transcriptional regulation of the human hepatic CYP3A4: identification of a new distal enhancer region responsive to CCAAT/enhancer-binding protein beta isoforms (liver activating protein and liver inhibitory protein). AB - CCAAT/enhancer-binding proteins (C/EBPs) are key transcription factors involved in the constitutive expression of several cytochrome P450 genes in the liver. Their concentration and activity change in several pathophysiological conditions. For instance, during inflammation, released cytokines induce repressive C/EBPbeta liver inhibitory protein (LIP), which antagonizes constitutive C/EBP transactivators [C/EBPalpha and C/EBPbeta-liver activating protein (LAP)], down regulating genes such as CYP3A4. However, the mechanism by which hepatic C/EBP factors modulate transcription of the CYP3A4 gene is not known. To elucidate the mechanism of action, we cotransfected luciferase reporter vectors, containing 5' flanking deletions of the CYP3A4 gene, along with expression vectors for C/EBPbeta-LAP, C/EBPbeta-LIP, and C/EBPalpha, in hepatic (HepG2) and nonhepatic (HeLa) cells. Analysis of the -3557 to -6954 base pair (bp) region demonstrated the existence of a 288-bp sequence at -5.95 kilobases (kb), which showed maximal response to C/EBPbeta-LAP ( approximately 30-fold increase in HepG2 cells). Coexpression of LAP with increasing amounts of LIP reduced the activating effect by approximately 70%. Site-directed mutagenesis of predicted C/EBPbeta binding sites demonstrated the presence of four functional C/EBPbeta-responsive motifs within this distal flanking region. Further experiments using chromatin immunoprecipitation proved the binding of endogenous C/EBPbeta to the -5.95 kilobase enhancer of the CYP3A4 gene in human hepatocytes. Expression of recombinant LAP and LIP by means of adenoviral vectors resulted in their binding to this region, which was followed by activation/repression of CYP3A4. Together, our results uncover a new distal enhancer site in the CYP3A4 gene where C/EBPbeta LAP binds and activates transcription, whereas the truncated form, C/EBPbeta-LIP, antagonizes LAP activity and causes gene repression. PMID- 15778454 TI - Centaurin-alpha 1, an ADP-ribosylation factor 6 GTPase activating protein, inhibits beta 2-adrenoceptor internalization. AB - The small GTP-binding protein ADP ribosylation factor 6 (ARF6) has recently been implicated in the internalization of G protein-coupled receptors (GPCRs), although its precise molecular mechanism in this process remains unclear. We have recently identified centaurin alpha(1) as a GTPase activating protein (GAP) for ARF6. In the current study, we characterized the effects of centaurin alpha(1) on the agonist-induced internalization of the beta(2)-adrenoceptor transiently expressed in human embryonic kidney (HEK) 293 cells. Using an enzyme-linked immunosorbent assay as well as confocal imaging of cells, we found that expression of centaurin alpha(1) strongly inhibited the isoproterenol-induced internalization of beta(2)-adrenoceptor. On the other hand, expression of functionally inactive versions of centaurin alpha(1), including an R49C mutant, which has no catalytic activity, and a double pleckstrin homology (PH) mutant (DM; R148C/R273C), which has mutations in both the PH domains of centaurin alpha(1), rendering it unable to translocate to the cell membrane, were unable to inhibit beta(2)-adrenoceptor internalization. In addition, a constitutively active version of ARF6, ARF6Q67L, reversed the ability of centaurin alpha(1) to inhibit beta(2)-adrenoceptor internalization. Finally, expression of centaurin alpha(1) also inhibited the agonist-induced internalization of beta(2) adrenoceptor endogenously expressed in HEK 293 cells, whereas the R49C and DM mutant versions of centaurin alpha(1) had no effect. Together, these data indicate that by acting as an ARF6 GAP, centaurin alpha(1) is able to switch off ARF6 and so inhibit its ability to mediate beta(2)-adrenoceptor internalization. Thus, ARF6 GAPs, such as centaurin alpha(1), are likely to play a crucial role in GPCR trafficking by modulating the activity of ARF6. PMID- 15778455 TI - Invasion of Lotus japonicus root hairless 1 by Mesorhizobium loti involves the nodulation factor-dependent induction of root hairs. AB - In many legumes, including Lotus japonicus and Medicago truncatula, susceptible root hairs are the primary sites for the initial signal perception and physical contact between the host plant and the compatible nitrogen-fixing bacteria that leads to the initiation of root invasion and nodule organogenesis. However, diverse mechanisms of nodulation have been described in a variety of legume species that do not rely on root hairs. To clarify the significance of root hairs during the L. japonicus-Mesorhizobium loti symbiosis, we have isolated and performed a detailed analysis of four independent L. japonicus root hair developmental mutants. We show that although important for the efficient colonization of roots, the presence of wild-type root hairs is not required for the initiation of nodule primordia (NP) organogenesis and the colonization of the nodule structures. In the genetic background of the L. japonicus root hairless 1 mutant, the nodulation factor-dependent formation of NP provides the structural basis for alternative modes of invasion by M. loti. Surprisingly, one mode of root colonization involves nodulation factor-dependent induction of NP-associated cortical root hairs and epidermal root hairs, which, in turn, support bacterial invasion. In addition, entry of M. loti through cracks at the cortical surface of the NP is described. These novel mechanisms of nodule colonization by M. loti explain the fully functional, albeit significantly delayed, nodulation phenotype of the L. japonicus ROOT HAIRLESS mutant. PMID- 15778456 TI - Aphid infestation causes different changes in carbon and nitrogen allocation in alfalfa stems as well as different inhibitions of longitudinal and radial expansion. AB - Alfalfa (Medicago sativa) stem elongation is strongly reduced by a pea aphid (Acyrthosiphon pisum Harris) infestation. As pea aphid is a phloem feeder that does not transmit virus or toxins, assimilate withdrawal is generally considered as the main mechanism responsible for growth reduction. Using a kinematic analysis, we investigated the spatial distributions of relative elemental growth rates of control and infested alfalfa stems. The water, carbon, and nitrogen contents per unit stem length were measured along the growth zone. Deposition rates and growth-sustaining fluxes were estimated from these patterns. Severe short-term aphid infestation (200 young adults over a 24-h period) induced a strong and synchronized reduction in rates of elongation and of water and carbon deposition. Reduced nitrogen content and associated negative nitrogen deposition rates were observed in some parts of the infested stems, especially in the apex. This suggested a mobilization of nitrogen from the apical part of the growth zone, converted from a sink tissue into a source tissue by aphids. Calculation of radial growth rates suggested that aphid infestation led to a smaller reduction in radial expansion than in elongation. Together with earlier observations of long-lasting effects of a short-term infestation, this supports the hypothesis that in addition to nutrient withdrawal, a thigmomorphogenesis-like mechanism is involved in the effect of aphid infestation on stem growth. PMID- 15778457 TI - RNA interference of soybean isoflavone synthase genes leads to silencing in tissues distal to the transformation site and to enhanced susceptibility to Phytophthora sojae. AB - Isoflavones are thought to play diverse roles in plant-microbe interactions and are also potentially important to human nutrition and medicine. Isoflavone synthase (IFS) is a key enzyme for the formation of the isoflavones. Here, we examined the consequences of RNAi silencing of genes for this enzyme in soybean (Glycine max). Soybean cotyledon tissues were transformed with Agrobacterium rhizogenes carrying an RNAi silencing construct designed to silence expression of both copies of IFS genes. Approximately 50% of emerging roots were transformed with the RNAi construct, and most transformed roots exhibited >95% silencing of isoflavone accumulation. Silencing of IFS was also demonstrated throughout the entire cotyledon (in tissues distal to the transformation site) both by high performance liquid chromatography analysis of isoflavones and by real-time reverse transcription-PCR. This distal silencing led to a nearly complete suppression of mRNA accumulation for both the IFS1 and IFS2 genes and of isoflavone accumulations induced by wounding or treatment with the cell wall glucan elicitor from Phytophthora sojae. Preformed isoflavone conjugates were not reduced in distal tissues, suggesting little turnover of these stored isoflavone pools. Distal silencing was established within just 5 d of transformation and was highly efficient for a 3- to 4-d period, after which it was no longer apparent in most experiments. Silencing of IFS was effective in at least two genotypes and led to enhanced susceptibility to P. sojae, disrupting both R gene-mediated resistance in roots and nonrace-specific resistance in cotyledon tissues. The soybean cotyledon system, already a model system for defense signal-response and cell-to-cell signaling, may provide a convenient and effective system for functional analysis of plant genes through gene silencing. PMID- 15778458 TI - Enhancing resistance to Sclerotinia minor in peanut by expressing a barley oxalate oxidase gene. AB - Sclerotinia minor Jagger is the causal agent of Sclerotinia blight, a highly destructive disease of peanut (Arachis hypogaea). Based on evidence that oxalic acid is involved in the pathogenicity of many Sclerotinia species, our objectives were to recover transgenic peanut plants expressing an oxalic acid-degrading oxalate oxidase and to evaluate them for increased resistance to S. minor. Transformed plants were regenerated from embryogenic cultures of three Virginia peanut cultivars (Wilson, Perry, and NC-7). A colorimetric enzyme assay was used to screen for oxalate oxidase activity in leaf tissue. Candidate plants with a range of expression levels were chosen for further analysis. Integration of the transgene was confirmed by Southern-blot analysis, and gene expression was demonstrated in transformants by northern-blot analysis. A sensitive fluorescent enzyme assay was used to quantify expression levels for comparison to the colorimetric protocol. A detached leaflet assay tested whether transgene expression could limit lesion size resulting from direct application of oxalic acid. Lesion size was significantly reduced in transgenic plants compared to nontransformed controls (65%-89% reduction at high oxalic acid concentrations). A second bioassay examined lesion size after inoculation of leaflets with S. minor mycelia. Lesion size was reduced by 75% to 97% in transformed plants, providing evidence that oxalate oxidase can confer enhanced resistance to Sclerotinia blight in peanut. PMID- 15778459 TI - Conservation of Arabidopsis flowering genes in model legumes. AB - The model plants Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa) have provided a wealth of information about genes and genetic pathways controlling the flowering process, but little is known about the corresponding pathways in legumes. The garden pea (Pisum sativum) has been used for several decades as a model system for physiological genetics of flowering, but the lack of molecular information about pea flowering genes has prevented direct comparison with other systems. To address this problem, we have searched expressed sequence tag and genome sequence databases to identify flowering-gene-related sequences from Medicago truncatula, soybean (Glycine max), and Lotus japonicus, and isolated corresponding sequences from pea by degenerate-primer polymerase chain reaction and library screening. We found that the majority of Arabidopsis flowering genes are represented in pea and in legume sequence databases, although several gene families, including the MADS-box, CONSTANS, and FLOWERING LOCUS T/TERMINAL FLOWER1 families, appear to have undergone differential expansion, and several important Arabidopsis genes, including FRIGIDA and members of the FLOWERING LOCUS C clade, are conspicuously absent. In several cases, pea and Medicago orthologs are shown to map to conserved map positions, emphasizing the closely syntenic relationship between these two species. These results demonstrate the potential benefit of parallel model systems for an understanding of flowering phenology in crop and model legume species. PMID- 15778460 TI - Overlaps in the transcriptional profiles of Medicago truncatula roots inoculated with two different Glomus fungi provide insights into the genetic program activated during arbuscular mycorrhiza. AB - Arbuscular mycorrhiza (AM) is a widespread symbiotic association between plants and fungal microsymbionts that supports plant development under nutrient-limiting and various stress conditions. In this study, we focused on the overlapping genetic program activated by two commonly studied microsymbionts in addition to identifying AM-related genes. We thus applied 16,086 probe microarrays to profile the transcriptome of the model legume Medicago truncatula during interactions with Glomus mosseae and Glomus intraradices and specified a total of 201 plant genes as significantly coinduced at least 2-fold, with more than 160 being reported as AM induced for the first time. Several hundred genes were additionally up-regulated during a sole interaction, indicating that the plant genetic program activated in AM to some extent depends on the colonizing microsymbiont. Genes induced during both interactions specified AM-related nitrate, ion, and sugar transporters, enzymes involved in secondary metabolism, proteases, and Kunitz-type protease inhibitors. Furthermore, coinduced genes encoded receptor kinases and other components of signal transduction pathways as well as AM-induced transcriptional regulators, thus reflecting changes in signaling. By the use of reporter gene expression, we demonstrated that one member of the AM-induced gene family encoding blue copper binding proteins (MtBcp1) was both specifically and strongly up-regulated in arbuscule-containing regions of mycorrhizal roots. A comparison of the AM expression profiles to those of nitrogen-fixing root nodules suggested only a limited overlap between the genetic programs orchestrating root endosymbioses. PMID- 15778461 TI - Tissue-specific localization of pea root infection by Nectria haematococca. Mechanisms and consequences. AB - Root infection in susceptible host species is initiated predominantly in the zone of elongation, whereas the remainder of the root is resistant. Nectria haematococca infection of pea (Pisum sativum) was used as a model to explore possible mechanisms influencing the localization of root infection. The failure to infect the root tip was not due to a failure to induce spore germination at this site, suppression of pathogenicity genes in the fungus, or increased expression of plant defense genes. Instead, exudates from the root tip induce rapid spore germination by a pathway that is independent of nutrient-induced germination. Subsequently, a factor produced during fungal infection and death of border cells at the root apex appears to selectively suppress fungal growth and prevent sporulation. Host-specific mantle formation in response to border cells appears to represent a previously unrecognized form of host-parasite relationship common to diverse species. The dynamics of signal exchange leading to mantle development may play a key role in fostering plant health, by protecting root meristems from pathogenic invasion. PMID- 15778462 TI - Inhibition of N2 fixation in soybean is associated with elevated ureides and amino acids. AB - Decreased N2 fixation in soybean (Glycine max) L. Merr. during water deficits has been associated with increases in ureides and free amino acids in plant tissues, indicating a potential feedback inhibition by these compounds in response to drought. We evaluated concentrations of ureides and amino acids in leaf and nodule tissue and the concurrent change in N2 fixation in response to exogenous ureides and soil-water treatments for the cultivars Jackson and KS4895. Exogenous ureides applied to the soil and water-deficit treatments inhibited N2 fixation by 85% to 90%. Mn fertilization increased the apparent catabolism of ureides in leaves and hastened the recovery of N2 fixation following exogenous ureide application for both cultivars. Ureides and total free amino acids in leaves and nodules increased during water deficits and coincided with a decline in N2 fixation for both cultivars. N2 fixation recovered to 74% to 90% of control levels 2 d after rewatering drought-stressed plants, but leaf ureides and total nodule amino acids remained elevated in KS4895. Asparagine accounted for 82% of the increase in nodule amino acids relative to well-watered plants at 2 d after rewatering. These results indicate that leaf ureides and nodule asparagine do not feedback inhibit N2 fixation. Compounds whose increase and decrease in concentration mirrored the decline and recovery of N2 fixation included nodule ureides, nodule aspartate, and several amino acids in leaves, indicating that these are potential candidate molecules for feedback inhibition of N2 fixation. PMID- 15778464 TI - Aphid resistance in Medicago truncatula involves antixenosis and phloem-specific, inducible antibiosis, and maps to a single locus flanked by NBS-LRR resistance gene analogs. AB - Aphids and related insects feed from a single cell type in plants: the phloem sieve element. Genetic resistance to Acyrthosiphon kondoi Shinji (bluegreen aphid or blue alfalfa aphid) has been identified in Medicago truncatula Gaert. (barrel medic) and backcrossed into susceptible cultivars. The status of M. truncatula as a model legume allows an in-depth study of defense against this aphid at physiological, biochemical, and molecular levels. In this study, two closely related resistant and susceptible genotypes were used to characterize the aphid resistance phenotype. Resistance conditions antixenosis since migratory aphids were deterred from settling on resistant plants within 6 h of release, preferring to settle on susceptible plants. Analysis of feeding behavior revealed the trait affects A. kondoi at the level of the phloem sieve element. Aphid reproduction on excised shoots demonstrated that resistance requires an intact plant. Antibiosis against A. kondoi is enhanced by prior infestation, indicating induction of this phloem-specific defense. Resistance segregates as a single dominant gene, AKR (Acyrthosiphon kondoi resistance), in two mapping populations, which have been used to map the locus to a region flanked by resistance gene analogs predicted to encode the CC-NBS-LRR subfamily of resistance proteins. This work provides the basis for future molecular analysis of defense against phloem parasitism in a plant model system. PMID- 15778463 TI - Partial reconstruction of flavonoid and isoflavonoid biosynthesis in yeast using soybean type I and type II chalcone isomerases. AB - Flavonoids and isoflavonoids are major plant secondary metabolites that mediate diverse biological functions and exert significant ecological impacts. These compounds play important roles in many essential physiological processes. In addition, flavonoids and isoflavonoids have direct but complex effects on human health, ranging from reducing cholesterol levels and preventing certain cancers to improving women's health. In this study, we cloned and functionally characterized five soybean (Glycine max) chalcone isomerases (CHIs), key enzymes in the phenylpropanoid pathway that produces flavonoids and isoflavonoids. Gene expression and kinetics analysis suggest that the soybean type I CHI, which uses naringenin chalcone as substrate, is coordinately regulated with other flavonoid specific genes, while the type II CHIs, which use a variety of chalcone substrates, are coordinately regulated with an isoflavonoid-specific gene and specifically activated by nodulation signals. Furthermore, we found that some of the newly identified soybean CHIs do not require the 4'-hydroxy moiety on the substrate for high enzyme activity. We then engineered yeast (Saccharomyces cerevisiae) to produce flavonoid and isoflavonoid compounds. When one of the type II CHIs was coexpressed with an isoflavone synthase, the enzyme catalyzing the first committed step of isoflavonoid biosynthesis, various chalcone substrates added to the culture media were converted to an assortment of isoflavanones and isoflavones. We also reconstructed the flavonoid pathway by coexpressing CHI with either flavanone 3beta-hydroxylase or flavone synthase II. The in vivo reconstruction of the flavonoid and isoflavonoid pathways in yeast provides a unique platform to study enzyme interactions and metabolic flux. PMID- 15778465 TI - Targeted proteomic analysis of 14-3-3 sigma, a p53 effector commonly silenced in cancer. AB - To comprehensively identify proteins interacting with 14-3-3 sigma in vivo, tandem affinity purification and the multidimensional protein identification technology were combined to characterize 117 proteins associated with 14-3-3 sigma in human cells. The majority of identified proteins contained one or several phosphorylatable 14-3-3-binding sites indicating a potential direct interaction with 14-3-3 sigma. 25 proteins were not previously assigned to any function and were named SIP2-26 (for 14-3-3 sigma-interacting protein). Among the 92 interactors with known function were a number of proteins previously implicated in oncogenic signaling (APC, A-RAF, B-RAF, and c-RAF) and cell cycle regulation (AJUBA, c-TAK, PTOV-1, and WEE1). The largest functional classes comprised proteins involved in the regulation of cytoskeletal dynamics, polarity, adhesion, mitogenic signaling, and motility. Accordingly ectopic 14-3-3 sigma expression prevented cellular migration in a wounding assay and enhanced mitogen activated protein kinase signaling. The functional diversity of the identified proteins indicates that induction of 14-3-3 sigma could allow p53 to affect numerous processes in addition to the previously characterized inhibitory effect on G2/M progression. The data suggest that the cancer-specific loss of 14-3-3 sigma expression by epigenetic silencing or p53 mutations contributes to cancer formation by multiple routes. PMID- 15778466 TI - Cooling the foot to prevent diabetic foot wounds: a proof-of-concept trial. AB - The etiology of neuropathic diabetic foot wounds can be summarized by the following formula: pressure x cycles of repetitive stress = ulceration. The final pathway to ulceration consists of an inflammatory response, leading to tissue breakdown. Mitigation of this response might reduce the risk of ulceration. This proof-of-concept trial evaluates whether simple cooling of the foot can safely reduce the time to thermal equilibrium after activity. After a 15-min brisk walk, the six nondiabetic volunteers enrolled were randomly assigned to receive either air cooling or a 10-min 55 degrees F cool water bath followed by air cooling. The process was then repeated with the intervention reversed, allowing subjects to serve as their own controls. There was a rise in mean +/- SD skin temperature after 15 min of activity versus preactivity levels (87.8 degrees +/- 3.9 degrees versus 79 degrees +/- 2.2 degrees F; P = .0001). Water cooling immediately brought the foot to a point cooler than preactivity levels for all subjects, whereas air cooling required an average of nearly 17 min to do so. Ten minutes of cooling required a mean +/- SD of 26.2 +/- 5.9 min to warm to preactivity levels. No adverse effects resulted from the intervention. We conclude that cooling the foot may be a safe and effective method of reducing inflammation and may serve as a prophylactic or interventional tool to reduce skin breakdown risk. PMID- 15778467 TI - Sinus tarsi denervation: clinical results. AB - Traumatic neuroma of the branches of the deep peroneal nerve that innervate the sinus tarsi can be the source of recalcitrant lateral ankle pain. That these nerves can be the source of the pain can be demonstrated by nerve blocks, and this pain can be surgically treated by resection of the appropriate branch of the deep peroneal nerve. This article documents the clinical results of this approach in 13 patients with sinus tarsi syndrome. At a minimum of 6 months postoperatively, 10 patients (77%) were completely pain-free, wore normal shoes, and had returned to work. Two patients (15%) had a small degree of residual pain but resumed usual activities and wore normal footwear. One patient had some pain relief but could not resume usual activities. We conclude that denervation of the sinus tarsi can relieve recalcitrant pain emanating from the sinus tarsi. This approach may reduce the need for subtalar fusion or evacuation procedures, including arthroereisis, thus avoiding their potential complications. Moreover, sinus tarsi denervation may allow the continued use of an arthroereisis implant in the presence of satisfactory objective findings, despite the subjective presence of postoperative pain. PMID- 15778468 TI - Use of the longitudinal arch angle to predict dynamic foot posture in walking. AB - To determine whether differences exist in the longitudinal arch angle based on sex or extremity, the longitudinal arch angle was assessed in 21 men and 21 women using a digital image of the medial aspect of each subject's feet. The image was obtained with the subject in relaxed standing posture and in maximum internal rotation of the lower leg. To determine whether the longitudinal arch angle could be used to predict dynamic foot posture during walking, 50 different subjects were asked to walk across a 6-m walkway while the medial aspect of each foot was videotaped. The longitudinal arch angle was digitized from digital images obtained at midstance for three walking trials. No differences in the longitudinal arch angle were found based on sex or extremity. The longitudinal arch angles obtained in the static positions of relaxed standing posture and maximum internal rotation were highly predictive of dynamic foot posture at midstance during walking. Relaxed standing posture and maximum internal rotation significantly contributed to explaining more than 90% of the variance associated with the longitudinal arch angle position at midstance during walking. These results validate use of the longitudinal arch angle as part of the foot and ankle physical examination. PMID- 15778469 TI - Management of hallux limitus with distraction of the first metatarsophalangeal joint. AB - This article describes a joint-preserving and joint-restoring procedure for the management of hallux limitus and hallux rigidus. The procedure uses a minirail external fixator to obtain distraction with or without arthrotomy of the first metatarsophalangeal joint. This procedure aims to restore joint function through elimination of the pathologic forces involved in hallux limitus and hallux rigidus. Both intrinsic and extrinsic muscular imbalances are reduced. Follow-up of 133 patients treated in this manner since 1997 demonstrates excellent long term results. PMID- 15778470 TI - Future of gait analysis: a podiatric medical perspective. AB - Despite the plethora of information on human gait analysis, its continued use as a clinical tool remains uncertain. Analysis of gait dysfunction has become integral to podiatric medical practice, and, like many specialized fields, it is rapidly changing to meet the needs of the future. Practice in the 21st century is predicated on the concept of multidisciplinary working approaches and a growing trend toward evidence-based practice, in which gait analysis could play a prominent role. This article provides a historical synopsis of instrumented gait analysis and its associated subcomponents and discusses the salient issues concerning its future role in podiatric medicine. PMID- 15778471 TI - Improved sensitivity in patients with peripheral neuropathy: effects of monochromatic infrared photo energy. AB - The medical records of 1,047 patients (mean age, 73 years) with established peripheral neuropathy were examined to determine whether treatment with monochromatic infrared photo energy was associated with increased foot sensitivity to the 5.07 Semmes-Weinstein monofilament. The peripheral neuropathy in 790 of these patients (75%) was due to diabetes mellitus. Before treatment with monochromatic infrared photo energy, of the ten tested sites (five on each foot), a mean +/- SD of 7.9 +/- 2.4 sites were insensitive to the 5.07 Semmes Weinstein monofilament, and 1,033 patients exhibited loss of protective sensation. After treatment, the mean +/- SD number of insensate sites on both feet was 2.3 +/- 2.4, an improvement of 71%. Only 453 of 1,033 patients (43.9%) continued to have loss of protective sensation after treatment. Therefore, monochromatic infrared photo energy treatment seems to be associated with significant clinical improvement in foot sensation in patients, primarily Medicare aged, with peripheral neuropathy. Because insensitivity to the 5.07 Semmes-Weinstein monofilament has been reported to be a major risk factor for diabetic foot wounds, the use of monochromatic infrared photo energy may be associated with a reduced incidence of diabetic foot wounds and amputations. PMID- 15778472 TI - A critical discussion of the use of antiseptics in acute traumatic wounds. AB - Approximately 10 million patients with traumatic wounds are treated in US emergency departments annually. The practice of wound cleansing or antiseptic management has a dichotomous history anchored in tradition and science. The merits of antiseptic fluid irrigation of traumatic wounds have received little scientific study. The purpose of this article is to critically evaluate the potential harm to patient outcome by the use of antiseptics on acute wounds. First, animal and cell culture data that describe the effects of topical antiseptics on wound healing are offered. Second, human case studies are presented to illustrate the potential harm of the indiscriminate use of antiseptics. Finally, data from previously published reviews are presented and evaluated for clinically based evidence to justify the current practice of antiseptic use in acute traumatic wounds. PMID- 15778473 TI - A simple technique for correcting footdrop: suspension tenodesis of the tibialis anterior tendon to the distal tibia. AB - Surgical correction of the footdrop deformity caused by various neuromuscular diseases is a time-honored technique, with most surgical procedures intended to correct the absence of active dorsiflexion and a concomitant cavovarus foot deformity. We describe suspension tenodesis of the tibialis anterior tendon to the distal tibia to correct the footdrop deformity. This technique is simpler to perform and more secure than transferring the distal attachment of a tendon to a new location in the foot itself or using tendon weaving techniques. In addition, compared with an ankle fusion, our technique allows for a certain amount of shock absorption during gait and is much less invasive, and a future ankle fusion could be performed if needed for full stability and definitive correction of the symptomatic footdrop. Finally, the technique we describe is easily reproducible and seems to remain stable over time. PMID- 15778474 TI - Unusually large cellular schwannoma of the foot. AB - This article presents a case of an unusually large cellular schwannoma of the foot. This uncommon lesion of the lower extremity is described with regard to clinical appearance, radiologic evaluation (including magnetic resonance imaging), histologic characteristics, and intraoperative findings. PMID- 15778475 TI - Diffuse pigmented villonodular synovitis: two cases involving the calcaneocuboid joint. AB - Pigmented villonodular synovitis is nonmalignant and nonmetastasizing, but it is locally destructive and can result in considerable disability through infiltration and involvement of surrounding soft tissues and bone. This article briefly describes the clinical picture of the diffuse form of pigmented villonodular synovitis and reports on two cases involving juxta-articular erosions of the calcaneocuboid joint. Treatment involved substantial curettage of bone and resection of infiltrated intrinsic musculature. PMID- 15778476 TI - Melorheostosis in a pediatric patient. AB - Melorheostosis is a nonhereditary and uncommon condition that can affect both adults and children. It can appear on radiographs as increased sclerosis on bones of the upper and lower extremities and may mimic other bony conditions such as osteopoikilosis, osteopetrosis, arthrogryposis multiplex congenita, and osteopathia striata. The sclerotic appearance can differ greatly between adults and children. The skin and subcutaneous tissues may be affected by fibrosis, resulting in contractures of joints and limbs that lead to deformities and limb length discrepancies. This article reviews the literature on melorheostosis and describes a case in a 10-year-old boy. PMID- 15778477 TI - Ulceration of pyoderma gangrenosum treated with negative pressure wound therapy. AB - Pyoderma gangrenosum is a skin disease characterized by wounds with blue-to purple undermined borders surrounding purulent necrotic bases. This article reports on a patient with a circumferential, full-thickness, and partially necrotic lower-extremity ulceration of unknown etiology. Results of laboratory tests and arterial and venous imaging studies were found to be within normal limits. The diagnosis of pyoderma gangrenosum was made on the basis of the histologic appearance of the wound tissue after biopsy as a diagnosis of exclusion. Negative pressure wound therapy was undertaken, which saved the patient's leg from amputation. Although negative pressure wound therapy has demonstrated efficacy in the treatment of chronic wounds in a variety of circumstances, this is the first documented use of this technique to treat an ulceration secondary to pyoderma gangrenosum. PMID- 15778478 TI - Onychoplasty with carbon dioxide laser matrixectomy for treatment of ingrown toenails. AB - Ingrown toenails are one of the most common pathologic conditions encountered in podiatric medical practice. Many methods of treatment for ingrown toenails have been used and studied, including chemical matrixectomies, surgical approaches, and CO(2) laser ablation. This study is a retrospective review of a new technique that consists of resection of the involved nail matrix using a No. 15 blade and controlled cauterization using a CO(2) laser. The technique was performed on 381 painful ingrown toenails, and all of the patients were followed up postoperatively for an average of 34 months. The results showed minimal pain, a low recurrence rate, rapid return to activity, and good cosmesis. PMID- 15778479 TI - Capsular interposition for the Keller bunionectomy with the use of soft-tissue anchors. AB - The Keller procedure has been used during the past century for the treatment of first metatarsophalangeal joint pathology. Many modifications to the procedure have been made, including interposition of the joint capsule into the first metatarsophalangeal joint space. Capsular interposition is often the most difficult step in performing the Keller bunionectomy. This article describes a new, simplified technique for capsular interposition with the use of a dorsal capsular flap and soft-tissue anchors. PMID- 15778481 TI - Internet resources for podiatric medical students: an update. AB - This article presents a selection of Internet resources covering most of the subject areas found in standard medical education curricula. Basic-sciences sites are emphasized, but clinical resources are also included. Sites were evaluated on the basis of their potential to enhance the learning process, provide practice questions or study guides for examinations, or aid in the preparation of papers. Podiatric medical students, residents, and practitioners who require a quick reference guide to sources covering the basic-science foundations of podiatric medicine or the clinical side of general medicine may find this article useful. PMID- 15778483 TI - Robin Goodfellow. PMID- 15778482 TI - Brucella osteomyelitis of the calcaneus. PMID- 15778484 TI - How pharmacogenomics will play a role in the management of asthma. PMID- 15778485 TI - PO2-dependent changes in intrinsic and extrinsic tongue muscle activities in the rat. AB - RATIONALE: Historically, respiratory-related research in sleep apnea has focused exclusively on the extrinsic tongue muscles (i.e., genioglossus, hyoglossus, and styloglossus). Until recently, the respiratory control and function of intrinsic tongue muscles (i.e., inferior and superior longitudinalis, transverses, and verticalis), which comprise the bulk of the tongue, were unknown. OBJECTIVES: The current study sought to determine if extrinsic and intrinsic tongue muscles are coactivated in conditions of hypoxemia comparable to that experienced by adults with obstructive sleep apnea. MEASUREMENTS: Esophageal pressure and EMG activity of an extrinsic (hyoglossus) and an intrinsic (superior longitudinal) tongue muscle were studied in anesthetized, tracheotomized, spontaneously breathing rats. Average EMG activity was compared in a control gas condition (Pa(O2), 160 +/- 12 mm Hg) and in mild isocapnic hypoxia (Pa(O2), 69 +/- 7.2 mm Hg), with and without brief (3-breath) airway occlusions, pre- and postbilateral vagotomy. MAIN RESULTS: (1) intrinsic and extrinsic tongue muscles are coactivated in mild hypoxia, (2) airway occlusion increased the activities of intrinsic retractor muscles in mild hypoxia, and (3) extrinsic retractor muscles have a steeper rate of rise of activity and an earlier burst onset relative to intrinsic retractor activities in mild hypoxia. CONCLUSIONS: These findings support our working hypothesis that airway patency is maintained not simply by activation of extrinsic tongue muscles but by the coactivation of intrinsic and extrinsic protrudor and retractor muscles. PMID- 15778486 TI - Rescue of the hypoplastic lung by prenatal cyclical strain. AB - We determined the effects of sustained and cyclical prenatal mechanical strain on the hypoplastic lung of the ovine model of congenital diaphragmatic hernia. Over a period of 4 weeks in late gestation, repeated cyclical tracheal occlusion for 23 hours with 1-hour release stimulated minimal growth, but promoted maturation with the development of a saccular lung. In contrast, a cycle consisting of 47 hours with 1-hour release induced optimal lung growth and morphologic maturation of the hypoplastic lung parenchyma. Sustained occlusion resulted in exaggerated lung growth, exceeding that of unaffected controls, and abnormal alveolar development. The extent of induction of lung growth by mechanical strain was inversely proportional to the number of alveolar type II cells remaining in the lung epithelium. These studies show that, although mechanical strain is capable of inducing lung growth and differentiation, cyclical strain is a prerequisite for normal development and that mechanically induced growth occurs at the expense of the alveolar type II cell. We conclude that cyclical strain may allow optimal alveolar development while maintaining a population of alveolar type II cells and may thus facilitate an improvement in postnatal lung function in infants with congenital diaphragmatic hernia. PMID- 15778487 TI - Pulmonary rehabilitation in chronic obstructive pulmonary disease. PMID- 15778488 TI - Dual role of vascular endothelial growth factor in experimental obliterative bronchiolitis. AB - Obliterative bronchiolitis (OB) is the major limitation for long-term survival of lung allograft recipients. We investigated the role of vascular endothelial growth factor (VEGF) in the development of OB in rat tracheal allografts. In nonimmunosuppressed allografts, VEGF mRNA and protein expression vanished in the epithelium and increased in smooth muscle cells and mononuclear inflammatory cells with progressive loss of epithelium and airway occlusion compared with syngeneic grafts. Intragraft VEGF overexpression by adenoviral transfer of a mouse VEGF(164) gene increased early epithelial cell proliferation and regeneration but increased microvascular remodeling and lymphangiogenesis and luminal occlusion by more than 50% compared with AdlacZ-treated allografts. Although VEGF receptor inhibition decreased early epithelial regeneration in noninfected allografts, it reduced microvascular remodeling, lymphangiogenesis, intragraft traffic of CD4(+) and CD8(+) T cells, and the degree of luminal occlusion. Simultaneous VEGF gene transfer and platelet-derived growth factor receptor inhibition with imatinib preserved respiratory epithelium and totally prevented luminal occlusion. In conclusion, our findings indicate that VEGF has a dual role in transplant OB. Our results suggest that VEGF may protect epithelial integrity. On the other hand, VEGF may enhance luminal occlusion by increasing the recruitment of mononuclear inflammatory cells with platelet-derived growth factor acting as a final effector molecule in this process. PMID- 15778489 TI - Fluctuations in end-expiratory lung volume during Cheyne-Stokes respiration. AB - We hypothesized that patients with Cheyne-Stokes respiration exhibit periodic increases in end-expiratory lung volume, mediated by changes in breath components, postinspiratory inspiratory muscle activity, or both. Calibrated inductive plethysmography revealed that 12 of 12 patients with Cheyne-Stokes respiration experienced increases in end-expiratory volume during hyperpnea: maximum 412 +/- 112 (SE) ml (range 75-1,543 ml). Compared with quiet breathing, the breath with largest increase in end-expiratory volume had larger tidal volume (867 +/- 107 vs. 567 +/- 38 ml, p < 0.01) and shorter expiratory time (1.25 +/- 0.11 vs. 1.66 +/- 0.15 seconds, p < 0.05). During decrescendo, the breath with largest decrease in end-expiratory volume had smaller tidal volume (p < 0.01) and longer expiratory time (p < 0.01). Cross-correlation of time series revealed that end-expiratory volume was related to both breath components (p < 0.0001). Bipolar needle electrodes revealed that scalene muscle activity at end-expiration was 50.7 +/- 14.0% higher at highest increase in lung volume than during preceding apnea (p < 0.05). Time series for scalene activity and end-expiratory volume were cross-correlated (p < 0.008). Increase in tonic scalene activity at end expiration, however, was equivalent during crescendo and decrescendo phases: 50.6 +/- 22.1 versus 42.0 +/- 12.9% (p = 0.48). In conclusion, patients with Cheyne Stokes respiration exhibit fluctuations in end-expiratory lung volume, primarily because of alterations in tidal volume and expiratory time rather than postinspiratory inspiratory muscle activity. PMID- 15778490 TI - Asthma: one hundred years of treatment and onward. AB - There have been four types of drug treatment of asthma that have been used over the past 100 years. Belladonna alkaloids, derived from the thorn-apple plant were used in 1905, and chemically synthesized entities in this class are still in use today. Western medicine began to use adrenergic stimulants approximately 100 years ago, but they were likely used in Asian medicine long before that. Systemic treatment with corticosteroids was introduced into the treatment of asthma in the mid-20th century; inhaled corticosteroids have been in use for over 35 years. The last 40 years have also seen the development of the first targeted asthma treatments: cromones, antileukotrienes, and anti-IgE. As we learn more of the biology of asthma, we anticipate that more effective targeted asthma treatments will be developed. PMID- 15778491 TI - Developmental regulation of p66Shc is altered by bronchopulmonary dysplasia in baboons and humans. AB - RATIONALE: The p66(Shc) adapter protein antagonizes mitogen-activated protein, or MAP, kinase, mediates oxidative stress, and is developmentally regulated in fetal mouse lungs. OBJECTIVES: To determine if p66(Shc) is similarly regulated in primates and in bronchopulmonary dysplasia (BPD), which results from oxidative injury to immature lungs. METHODS: Normal and injured lungs from humans and baboons were evaluated by Western analysis and immunohistochemistry. MEASUREMENTS AND MAIN RESULTS: In baboons, p66(Shc) decreased 80% between 125 and 175 days' gestation (p = 0.025), then doubled after term delivery at 185 days (p = 0.0013). In the hyperoxic 140-day fetal baboon BPD model, p66(Shc) expression persisted, and its localization shifted from the epithelium of gestational controls to the mesenchyme of diseased lungs, coincident with expression of proliferating cell nuclear antigen and cleaved poly(adenyl ribose) polymerase, a marker of apoptosis. Treatment with the antibombesin antibody 2A11 attenuated BPD, reduced cell proliferation, increased p66(Shc) expression 10.5-fold, and preserved epithelial p66(Shc) localization. p66(Shc) also decreased during normal human lung development, falling 87% between 18 and 24 weeks' gestation (p = 0.02). p66(Shc) was expressed throughout 18-week human lungs, became restricted to scattered epithelial cells by 24 weeks, and localized to isolated mesenchymal cells after term delivery. In contrast, p66(Shc) remained prominent in the epithelium of lungs with acute injury or mild BPD, and in the mesenchyme of lungs with severe disease. p66(Shc) localized to tissues expressing proliferating cell nuclear antigen and cleaved poly(adenyl ribose) polymerase. CONCLUSIONS: p66(Shc) expression, cell proliferation, and apoptosis are concomitantly altered during lung development and in BPD. PMID- 15778492 TI - Induction of CXCL5 during inflammation in the rodent lung involves activation of alveolar epithelium. AB - The lung is continuously exposed to bacteria and their products, and has developed a complex defense mechanism, including neutrophil recruitment. In mice, keratinocyte cell-derived chemokine and macrophage inflammatory protein-2 are the major chemokines for neutrophil recruitment into the lung. We have previously described a role for C-X-C chemokine (CXCL5) in neutrophil trafficking during lipopolysaccharide (LPS)-induced lung inflammation in mice. The aims of the present study were to identify the cellular origin of CXCL5 and to determine the signaling cascades that regulate its expression in the lung during LPS-induced inflammation and in isolated LPS-stimulated CXCL5-expressing cells. Our immunohistochemical analysis indicates that alveolar epithelial type II (AEII) cells are the primary source of CXCL5 in the rodent lung. These in vivo observations were confirmed with primary AEII cells. In addition, our data indicate that the Toll-like receptor 4 (TLR4) signaling cascade involving TLR4, myeloid differentiation factor 88, and Toll-IL-1R domain-containing adapter protein is required to induce CXCL5 expression in the lung. Furthermore, p38 and c-Jun N-terminal kinases are involved in lung CXCL5 expression. Similarly, TLR4, and p38 and c-Jun N-terminal kinases, are associated with LPS-induced CXCL5 expression in AEII cells. These novel observations demonstrate that activation of AEII cells via TLR4-dependent signaling is important for the production of CXCL5 in the lung exposed to LPS. PMID- 15778493 TI - Resident Th1-like effector memory cells in pulmonary recall responses to Mycobacterium tuberculosis. AB - We recently described a model of Th1 recall responses based on segmental antigen challenge with purified protein derivative of Mycobacterium tuberculosis (PPD). Bronchoscopic instillation of 0.5 tuberculin units of PPD resulted in localized lymphocytic inflammation in PPD-positive subjects only. Recruited lymphocytes were predominantly CD4+ and were enriched for cells capable of PPD-specific interferon (IFN)-gamma production. In the current study, we investigated the mechanisms by which this localized recall response is mobilized. Bronchoscopic PPD challenge of skin test-positive subjects resulted in the production of CXCR3 ligands IFN-gamma-inducible protein (IP)-10 and monokine induced by IFN-gamma (Mig), but not of CCR5 ligands macrophage inflammatory protein-1alpha and regulated-upon activation, normal T-cell expressed and secreted, whereas skin test-negative subjects produced none of these chemokines. Baseline bronchoalveolar lavage (BAL) cells of skin test-positive subjects produced IP-10 and Mig in response to in vitro stimulation as well. Because IP-10 and Mig are IFN-gamma-inducible chemokines, these findings suggested that chemokine responses to PPD were facilitated by resident memory cells of the lung. Further studies confirmed that baseline BAL lymphocytes of PPD-positive subjects produce IFN gamma in response to PPD, and that, compared with peripheral blood, BAL cells are preferentially enriched for PPD-specific lymphocytes. This IFN-gamma production is predominantly a function of CD4+ T cells that display the CD45RO+/CCR7- surface phenotype characteristic of effector memory cells. PMID- 15778494 TI - The role of the extracellular domain in the biology of the coxsackievirus and adenovirus receptor. AB - The Coxsackievirus B and Adenovirus Receptor (CAR) plays a dual role as a homotypic junctional adhesion protein and as a viral receptor. CAR is a transmembrane protein and a member of the Immunoglobulin (Ig) superfamily with two extracellular Ig-like domains. The most distal Ig-like domain (D1) mediates the homophilic interaction and is also responsible for the high-affinity binding of the adenovirus (Ad) fiber protein. Currently, no activity has been ascribed to the proximal Ig-like domain (D2). To further understand the function of the extracellular domain in the biological activities of CAR, we created extracellular deletion mutants and evaluated cellular localization, adhesion, and viral infection. Deletion of any segment of the extracellular domain results in loss of adhesion and mislocalization as explained by a model, termed "diffusion trapping," that suggests adhesion is the driving force in junctional localization. Loss of junctional localization and adhesion was particularly apparent in polarized human airway epithelia, where mutant CAR expression was basolateral but not limited to the lateral junctions between cells. Surprisingly, the D2 domain was required for adenovirus fiber-knob binding and infection. In summary, the entire extracellular domain of CAR is of vital importance to the biology of this highly conserved and important protein. PMID- 15778495 TI - A surfactant protein C precursor protein BRICHOS domain mutation causes endoplasmic reticulum stress, proteasome dysfunction, and caspase 3 activation. AB - BRICHOS is a domain found in several proteins consisting of approximately 100 amino acids with sequence and structural similarities. Mutations in BRICHOS domain have been associated with both degenerative and proliferative diseases in several nonpulmonary organs, although the pathogenic mechanisms are largely undefined. Recently, several mutations in surfactant protein C (SP-C) mapping to the BRICHOS domain located within the proprotein (proSP-C) have been linked to interstitial lung diseases. In vitro expression of one of these BRICHOS mutants, the exon 4 deletion (hSP-CDeltaexon4), promotes a dominant-negative perinuclear aggregation of the protein. The present study characterizes the trafficking behavior and pathogenic consequences resulting from hSP-CDeltaexon4 expression. Time-lapse and co-localization microscopy studies demonstrated enhanced green fluorescent protein (EGFP)/hSP-CDeltaexon4 expression in calnexin-positive (endoplasmic reticulum [ER]) compartment with subsequent time- and concentration dependent development of ubiquitinated perinuclear inclusion bodies followed by apoptosis. Compared with controls, EGFP/hSP-CDeltaexon4 promoted upregulation of multiple ER stress species, activated caspase 3, and induced annexin V binding. Furthermore, in GFP-u cells, hSP-CDeltaexon4 directly inhibited proteasome activity. These results support a model whereby proSP-C BRICHOS mutations induce a dynamic toxic gain-of-function, causing apoptotic cell death both by early ER accumulation leading to an exaggerated unfolded protein response and by enhanced deposition of cellular aggregates associated with proteasome dysfunction. PMID- 15778496 TI - Interleukin-2-inducible T cell kinase regulates mast cell degranulation and acute allergic responses. AB - Bruton's tyrosine kinase (Btk) is thought to positively regulate mast cell activation, implying a role in allergic responses. We have compared acute and late phase allergic airway reactions in mice lacking either Btk or interleukin-2 inducible T cell kinase (Itk), another Tec kinase expressed in mast cells. Btk(-/ ) mice showed minor protection against allergic symptoms when challenged with allergen via the airways. In sharp contrast, both acute and late phase inflammatory allergic responses were markedly reduced in Itk(-/-) mice. Notably, airway mast cell degranulation in Itk(-/-) mice was severely impaired, despite wild-type levels of allergen-specific IgE and IgG1. The degranulation defect was confirmed in DNP-conjugated human serum albumin-challenged mice passively sensitized with anti-DNP IgE antibodies, and was also observed after direct G protein stimulation with the mast cell secretagogue c48/80. Moreover, late phase inflammatory changes, including eosinophilia, lymphocyte infiltration, and Th2 cytokine production in the lungs, was eliminated in Itk(-/-) mice. Collectively, our data suggest a critical role of Itk in airway mast cell degranulation in vivo that together with an impaired T cell response prevents the development of both acute and late phase inflammatory allergic reactions. PMID- 15778497 TI - Attenuation of endotoxin-induced acute lung injury by the Rho-associated kinase inhibitor, Y-27632. AB - A small GTPase, Rho, plays key roles in cell adhesion, motility, and contraction after stimulation. Among Rho effectors isolated, the family of Rho-associated coiled-coil-forming protein kinases (ROCK) is implicated in Rho-mediated cell adhesion and smooth muscle contraction. The effect of a specific inhibitor of ROCK, Y-27632, was evaluated in a murine model of acute lung injury induced by intravenous injection of Escherichia coli endotoxin (lipopolysaccharide [LPS]). Lung edema was evaluated by measuring extravascular leakage of radio-labeled serum albumin, and neutrophil emigration into the lung parenchyma by morphometric observation and measuring myeloperoxidase activity. Pretreatment with Y-27632 attenuated both lung edema and neutrophil emigration after LPS. We also measured albumin transfer through cultured endothelial cell monolayers on a porous filter. Tumor necrosis factor-alpha significantly increased albumin transfer, which was attenuated by pretreatment with Y-27632. Fluorescence microscopy revealed that morphologic changes in endothelial cells induced by tumor necrosis factor-alpha were inhibited by Y-27632. In contrast, the increased fraction of neutrophils with polymerized actin after formyl-methionyl-leucyl-phenylalanine was not altered by Y-27632. These data suggest that ROCK may play an important role in the pathogenesis of LPS-induced lung injury and that ROCK inhibition could attenuate cytoskeletal rearrangement of endothelial cells, leading to decreased neutrophil emigration into the lung parenchyma. PMID- 15778499 TI - Mint represses transactivation of the type II collagen gene enhancer through interaction with alpha A-crystallin-binding protein 1. AB - Collagen type II is an extracellular matrix protein important for cartilage and bone formation, and its expression is controlled by multiple cis- and trans acting elements, including the zinc finger transcription factor alpha A crystallin-binding protein 1 (CRYBP1). Here we show that MSX2-interacting nuclear target protein (MINT), a conserved transcriptional repressor, associates with CRYBP1 and negatively regulates the transactivation of the collagen type II gene (Col2a1) enhancer. We identified CRYBP1 as a binding partner of MINT by screening a mouse embryonic cDNA library using the yeast two-hybrid system. We demonstrated that the C terminus of MINT interacts with the C terminus of CRYBP1 using the mammalian cell two-hybrid assay, glutathione S-transferase pull-down, and co immunoprecipitation analyses. Furthermore, MINT and CRYBP1 form a complex on the Col2a1 enhancer, as shown by chromatin immunoprecipitation and gel shift assays. In the presence of CRYBP1, overexpression of MINT or its C-terminal fragment in cells repressed a reporter construct driven by the Col2a1 enhancer elements. This transcription repression is dependent on histone deacetylase, the main co repressor recruited by MINT. The present study shows that MINT is involved in CRYBP1-mediated Col2a1 gene repression and may play a role in regulation of cartilage development. PMID- 15778498 TI - Proline-rich tyrosine kinase 2 (Pyk2) mediates vascular endothelial-cadherin based cell-cell adhesion by regulating beta-catenin tyrosine phosphorylation. AB - Vascular endothelial-cadherin (VE-cadherin) controls endothelial cell-cell adhesion and preserves endothelial integrity. In order to maintain endothelial barrier function, VE-cadherin function is tightly regulated through mechanisms that involve protein phosphorylation and cytoskeletal dynamics. Here, we show that loss of VE-cadherin function results in intercellular gap formation and a drop in electrical resistance of monolayers of primary human endothelial cells. Detailed analysis revealed that loss of endothelial cell-cell adhesion, induced by VE-cadherin-blocking antibodies, is preceded by and dependent on a rapid activation of Rac1 and increased production of reactive oxygen species. Moreover, VE-cadherin-associated beta-catenin is tyrosine-phosphorylated upon loss of cell cell contact. Finally, the redox-sensitive proline-rich tyrosine kinase 2 (Pyk2) is activated and recruited to cell-cell junctions following the loss of VE cadherin homotypic adhesion. Conversely, the inhibition of Pyk2 activity in endothelial cells by the expression of CRNK (CADTK/CAKbeta-related non-kinase), an N-terminal deletion mutant that acts in a dominant negative fashion, not only abolishes the increase in beta-catenin tyrosine phosphorylation but also prevents the loss of endothelial cell-cell contact. These results implicate Pyk2 in the reduced cell-cell adhesion induced by the Rac-mediated production of ROS through the tyrosine phosphorylation of beta-catenin. This signaling is initiated upon loss of VE-cadherin function and is important for our insight in the modulation of endothelial integrity. PMID- 15778500 TI - Biosynthesis of UDP-N-acetyl-L-fucosamine, a precursor to the biosynthesis of lipopolysaccharide in Pseudomonas aeruginosa serotype O11. AB - UDP-N-acetyl-L-fucosamine is a precursor to l-fucosamine in the lipopolysaccharide of Pseudomonas aeruginosa serotype O11 and the capsule of Staphylococcus aureus type 5. We have demonstrated previously the involvement of three enzymes, WbjB, WbjC, and WbjD, in the biosynthesis of UDP-2-acetamido-2,6 dideoxy-L-galactose or UDP-N-acetyl-L-fucosamine (UDP-l-FucNAc). An intermediate compound from the coupled-reaction of WbjB-WbjC with the initial substrate UDP-2 acetamido-2-deoxy-alpha-D-glucose or UDP-N-acetyl-D-glucosamine (UDP-GlcNAc) was purified, and the structure was determined by NMR spectroscopy to be UDP-2 acetamido-2,6-dideoxy-L-talose (UDP-L-PneNAc). WbjD could then convert this intermediate into a new product with the same mass, consistent with a C-2 epimerization reaction. Those results led us to propose a pathway for the biosynthesis of UDP-L-FucNAc; however, the exact enzymatic activity of each of these proteins has not been defined. Here, we describe a fast protein liquid chromatography (FPLC)-based anion-exchange procedure, which allowed the separation and purification of the products of C-2 epimerization due to WbjD. Also, the application of a cryogenically cooled probe in NMR spectrometry offers the greatest sensitivity for determining the structures of minute quantities of materials, allowing the identification of the final product of the pathway. Our results showed that WbjB is bifunctional, catalyzing firstly C-4, C-6 dehydration and secondly C-5 epimerization in the reaction with the substrate UDP-D-GlcNAc, producing two intermediates. WbjC is also bifunctional, catalyzing C-3 epimerization of the second intermediate followed by reduction at C-4. The FPLC based procedure provided good resolution of the final product of WbjD reaction from its epimer/substrate UDP-l-PneNAc, and the use of the cryogenically cooled probe in NMR revealed unequivocally that the final product is UDP-L-FucNAc. PMID- 15778501 TI - JNK1 and JNK2 oppositely regulate p53 in signaling linked to apoptosis triggered by an altered fibronectin matrix: JNK links FAK and p53. AB - The extracellular matrix regulates many cellular processes, including survival, and alterations in the matrix or in matrix survival signals can trigger apoptosis. Previously, we showed that an altered fibronectin matrix triggers apoptosis in primary cells via a novel pathway regulated by transcriptionally mediated decreases in p53 and c-Myc levels. Here we report that this apoptotic mechanism is propagated by decreased phosphorylation of focal adhesion kinase (FAK), which is linked to increased phosphorylation of c-Jun N-terminal kinase (JNK) and to decreased levels of p53. FAK is physically and spatially linked to JNK and p53, which relocalize from the nucleus to the cell membrane to mediate this interaction. Further, p53 participates in a feedback mechanism with JNK to regulate this apoptotic process and is oppositely regulated by JNK1 and JNK2. PMID- 15778502 TI - P2Y2 nucleotide receptors enhance alpha-secretase-dependent amyloid precursor protein processing. AB - The amyloid precursor protein (APP) is proteolytically processed by beta- and gamma-secretases to release amyloid beta, the main component in senile plaques found in the brains of patients with Alzheimer disease. Alternatively, APP can be cleaved within the amyloid beta domain by alpha-secretase releasing the non amyloidogenic product sAPP alpha, which has been shown to have neuroprotective properties. Several G protein-coupled receptors are known to activate alpha secretase-dependent processing of APP; however, the role of G protein-coupled nucleotide receptors in APP processing has not been investigated. Here it is demonstrated that activation of the G protein-coupled P2Y2 receptor (P2Y2R) subtype expressed in human 1321N1 astrocytoma cells enhanced the release of sAPP alpha in a time- and dose-dependent manner. P2Y2 R-mediated sAPP alpha release was dependent on extracellular calcium but was not affected by 1,2-bis(2 aminophenoxy)ethane-N,N,N,-trimethylammonium salt, an intracellular calcium chelator, indicating that P2Y2R-stimulated intracellular calcium mobilization was not involved. Inhibition of protein kinase C (PKC) with GF109203 or by PKC down regulation with phorbol ester pre-treatment had no effect on UTP-stimulated sAPP alpha release, indicating a PKC-independent mechanism. U0126, an inhibitor of the mitogen-activated protein kinase pathway, partially inhibited sAPPalpha release by UTP, whereas inhibitors of Src-dependent epidermal growth factor receptor transactivation by P2Y2Rs had no effect. The metalloprotease inhibitors phenanthroline and TAPI-2 and the furin inhibitor decanoyl-Arg-Val-Lys-Arg chloromethylketone also diminished UTP-induced sAPP alpha release. Furthermore, small interfering RNA silencing of an endogenous adamalysin, ADAM10 or ADAM17/TACE, partially suppressed P2Y2R-activated sAPP alpha release, whereas treatment of cells with both ADAM10 and ADAM17/TACE small interfering RNAs completely abolished UTP-activated sAPP alpha release. These results may contribute to an understanding of the non-amyloidogenic processing of APP. PMID- 15778504 TI - Novel heparan sulfate structures revealed by monoclonal antibodies. AB - The sulfated glycosaminoglycan heparan sulfate (HS) is found ubiquitously on cell surfaces, in the extracellular matrix, and intracellularly as HS proteoglycans. Because of the structural heterogeneity of HS, tissue-derived HS preparations represent a mixture of HS chains originating from different cell types and tissue loci. Monoclonal anti-HS antibodies have been employed to detect the localization of specific HS epitopes in tissues, but limited information has been available on the saccharide structures recognized by the antibodies. We have studied the saccharide epitope structures of four anti-HS antibodies, HepSS1, JM13, JM403, and 10E4, which all recognize distinct HS species as demonstrated by different patterns of immunoreactivity upon staining of embryonic rat and adult human tissues. The epitopes recognized by JM13 and HepSS1 were found almost exclusively in basement membrane HS, whereas JM403 and 10E4 reacted also with cell-associated HS species. The binding of HepSS1, JM403, and 10E4 to HS was dependent on the GlcN N-substitution of the polysaccharide rather than O-sulfation. HepSS1 thus interacted with N-sulfated HS domains, JM403 binding was critically dependent on N-unsubstituted GlcN residues, and 10E4 bound to "mixed" HS domains containing both N-acetylated and N-sulfated disaccharide units. By contrast, JM13 binding seemed to require the presence of 2-O-sulfated glucuronic acid residues. PMID- 15778503 TI - Sclerostin binds to LRP5/6 and antagonizes canonical Wnt signaling. AB - The loss of the SOST gene product sclerostin leads to sclerosteosis characterized by high bone mass. In this report, we found that sclerostin could antagonize canonical Wnt signaling in human embryonic kidney A293T cells and mouse osteoblastic MC3T3 cells. This sclerostin-mediated antagonism could be reversed by overexpression of Wnt co-receptor low density lipoprotein receptor-related protein (LRP) 5. In addition, we found that sclerostin bound to LRP5 as well as LRP6 and identified the first two YWTD-EGF repeat domains of LRP5 as being responsible for the binding. Although these two repeat domains are required for transduction of canonical Wnt signals, canonical Wnt did not appear to compete with sclerostin for binding to LRP5. Examination of the expression of sclerostin and Wnt7b, an autocrine canonical Wnt, during primary calvarial osteoblast differentiation revealed that sclerostin is expressed at late stages of osteoblast differentiation coinciding with the expression of osteogenic marker osteocalcin and trailing after the expression of Wnt7b. Given the plethora of evidence indicating that canonical Wnt signaling stimulates osteogenesis, we believe that the high bone mass phenotype associated with the loss of sclerostin may be attributed, at least in part, to an increase in canonical Wnt signaling resulting from the reduction in sclerostin-mediated Wnt antagonism. PMID- 15778505 TI - Identification, evolution, and regulation of expression of Guinea pig trappin with an unusually long transglutaminase substrate domain. AB - Trappins are found in human, bovine, hippopotamus, and members of the pig family, but not in rat and mouse. To clarify the evolution of the trappin genes and the functional significance of their products, we isolated the trappin gene in guinea pig, a species belonging to a rodent family distinct from rat and mouse. Guinea pig trappin was confirmed to encode the same domain structure as trappin, consisting of a signal sequence, an extra large transglutaminase substrate domain, and a whey acidic protein motif. Northern blot analysis and in situ hybridization histochemistry as well as immunohistochemistry demonstrated that guinea pig trappin is expressed solely in the secretory epithelium of the seminal vesicle and that its expression is androgen-dependent. We confirmed that guinea pig trappin is cross-linked by prostate transglutaminase and that the whey acidic protein motif derived from guinea pig trappin has an inhibitory activity against leukocyte elastase. Genome sequence analysis showed that guinea pig trappin belongs to the family of REST (rapidly evolving seminal vesicle transcribed) genes. PMID- 15778506 TI - Fluid shear stress synergizes with insulin-like growth factor-I (IGF-I) on osteoblast proliferation through integrin-dependent activation of IGF-I mitogenic signaling pathway. AB - This study tested the hypothesis that shear stress interacts with the insulin like growth factor-I (IGF-I) pathway to stimulate osteoblast proliferation. Human TE85 osteosarcoma cells were subjected to a steady shear stress of 20 dynes/cm(2) for 30 min followed by 24-h incubation with IGF-I (0-50 ng/ml). IGF-I increased proliferation dose-dependently (1.5-2.5-fold). Shear stress alone increased proliferation by 70%. The combination of shear stress and IGF-I stimulated proliferation (3.5- to 5.5-fold) much greater than the additive effects of each treatment alone, indicating a synergistic interaction. IGF-I dose-dependently increased the phosphorylation level of Erk1/2 by 1.2-5.3-fold and that of IGF-I receptor (IGF-IR) by 2-4-fold. Shear stress alone increased Erk1/2 and IGF-IR phosphorylation by 2-fold each. The combination treatment also resulted in synergistic enhancements in both Erk1/2 and IGF-IR phosphorylation (up to 12- and 8-fold, respectively). Shear stress altered IGF-IR binding only slightly, suggesting that the synergy occurred primarily at the post-ligand binding level. Recent studies have implicated a role for integrin in the regulation of IGF-IR phosphorylation and IGF-I signaling. To test whether the synergy involves integrin-dependent mechanisms, the effect of echistatin (a disintegrin) on proliferation in response to shear stress +/- IGF-I was measured. Echistatin reduced basal proliferation by approximately 60% and the shear stress-induced mitogenic response by approximately 20%. It completely abolished the mitogenic effect of IGF-I and that of the combination treatment. Shear stress also significantly reduced the amounts of co-immunoprecipitated SHP-2 and -1 with IGF IR, suggesting that the synergy between shear stress and IGF-I in osteoblast proliferation involves integrin-dependent recruitment of SHP-2 and -1 away from IGF-IR. PMID- 15778507 TI - Pulmonary defense and the human cathelicidin hCAP-18/LL-37. AB - Antimicrobial peptides form an important component of the innate immune system. The cathelicidin family, a key member of the antimicrobial peptide defenses, has been highly conserved throughout evolution. Though widespread in mammals, there is currently only one identified human example, hCAP-18/LL-37. The cathelicidins have been found to have multiple functions, in addition to their known antimicrobial and lipopolysaccharide-neutralizing effects. As a result, they profoundly affect both innate and adaptive immunity. Currently, antimicrobial peptides are being evaluated as therapeutic drugs in disease states as diverse as oral mucositis, cystic fibrosis, and septic shock. One such peptide, the cathelicidin hCAP-18/LL-37, is reviewed in detail in the context of its role in lung physiology and defense. PMID- 15778508 TI - Understanding sex biases in immunity: effects of estrogen on the differentiation and function of antigen-presenting cells. AB - The initiation and perpetuation of innate and adaptive immunity is dependent on the ability of professional antigen-presenting cells (APCs) to sense inflammatory stimuli; produce cytokines; and internalize, degrade, and present antigens via surface major histocompatibility complex (MHC) molecules. Dendritic cells (DCs), macrophages, and B lymphocytes express estrogen receptors, indicating that the steroid sex hormone estrogen might directly modulate the function of these cells during immune responses. Sex-specific parameters of immune function have been identified during autoimmunity and the pathogenesis of infectious disease, which show sex biases in their incidence and manifestation; female immunity also varies as estrogen levels change. In this article, we summarize studies that demonstrate effects of estrogen on the differentiation or function of APCs in model in vitro systems, or under circumstances of natural or imposed variation in estrogen levels in vivo. PMID- 15778510 TI - CD19 function in central and peripheral B-cell development. AB - Although the B-cell antigen receptor (BCR) factors most prominently in the maintenance and differentiation of mature B cells, it is now appreciated that co receptor molecules can positively or negatively modulate signals through the BCR. Co-receptors are functionally defined as modifiers of BCR engagement and signal transduction, and are distinct from other accessory molecules that act independently to regulate B-cell growth. The co-receptor CD19 functions to augment signals by the pre-BCR/BCR and in doing so can modulate B-cell fate decisions at multiple stages of development. In mature B cells, CD19 also associates with complement receptor 2 (CR2/CD21) and is pivotal for transducing signals induced by co-recognition of complement C3d-fixed antigens by the BCR and CD21. In this article, we focus on recent progress in the understanding of CD19 function through the characterization of mouse models that relate in vivo function to biochemical properties of CD19. PMID- 15778511 TI - A biological context for the self-nonself discrimination and the regulation of effector class by the immune system. AB - An effective immune response to an antigen requires two sets of decisions: Decision 1, the sorting of the repertoire, and Decision 2, the regulation of effector class. The repertoire, because it is somatically generated, large, and random, must be sorted by a somatic mechanism that subtracts those specificities (anti-self) that, if expressed, would debilitate the host, leaving a residue (anti-nonself) that, if not expressed, would result in the death of the host by infection. The self-nonself discrimination is the metaphor used to describe Decision 1, the sorting of the repertoire. In order to be functional, the sorted repertoire must be coupled to a set of biodestructive and ridding effector functions, such that the response to each antigen is treated in a coherent and independent manner. Although a reasonably complete framework for Decision 1 exists, Decision 2 lacks conceptualization. The questions that must be considered to arrive at a proper framework are posed. It should be emphasized that manipulation at the level of Decision 2 is where clinical applications are likely to be found. PMID- 15778509 TI - Metabolic mechanisms of tumor resistance to T cell effector function. AB - Established tumors develop ways to elude destruction by the host immune system. Recent work has revealed that tumors can take advantage of the generation of metabolic dysregulation to inhibit immune responses. Effector T-cell functions are particularly sensitive to nutrient availability in the tumor microenvironment. In this review, we highlight experimental data supporting the importance of glucose, oxygen, tryptophan, and arginine for optimal T-cell function, and the mechanisms by which these nutrients may become depleted in the tumor microenvironment. These observations provide a conceptual framework for modulating metabolic features of the T cell-tumor interaction, toward the end of promoting more effective immune-mediated tumor destruction in vivo. PMID- 15778512 TI - The mdr1a-/- mouse model of spontaneous colitis: a relevant and appropriate animal model to study inflammatory bowel disease. AB - There are many types of colitis models in animals that researchers use to elucidate the mechanism of action of human inflammatory bowel disease (IBD). These models are also used to test novel therapeutics and therapeutic treatment regimens. Here, we will review the characteristics of the mdr1a -/- model of spontaneous colitis that we believe make this model an important part of the IBD researcher's toolbox. We will also share new data that will reinforce the fact that this model is relevant in the study of IBD. Mdr1a -/- mice lack the murine multiple drug resistance gene for P-glycoprotein 170 that is normally expressed in multiple tissues including intestinal epithelial cells. These mice spontaneously develop a form of colitis at around 12 wk of age. The fact that the complexity of this model mirrors the complexity of disease in humans, as well as recent literature that links MDR1 polymorphisms in humans to Crohn's Disease and Ulcerative Colitis, makes this an appropriate animal model to study. PMID- 15778513 TI - Are we really on the right TRAIL? PMID- 15778515 TI - Complete rectal prolapse--quo vadis? PMID- 15778516 TI - Hepatitis B virus genotyping: will it stand the test of time? PMID- 15778517 TI - Postoperative pulmonary function in laparoscopic versus open cholecystectomy: prospective, comparative study. AB - BACKGROUND: Pulmonary complications remain a leading cause of morbidity after major abdominal operations. OBJECTIVE: To compare pulmonary function and the frequency of pulmonary complications after laparoscopic cholecystectomy (LC) and open cholecystectomy (OC). METHODS: Fifty-five patients with symptomatic gallstone disease undergoing elective cholecystectomy (LC 40, OC 15) under general anesthesia were evaluated using pulmonary function tests (forced vital capacity [FVC], forced expiratory volume at 1 second [FEV1], and forced expiratory flow at 25% to 75% [FEF25% -75%], chest X-ray and pulse oximetry before and after surgery. RESULTS: FVC, FEV1 and FEF25% -75% decreased by 21.5%, 21.2% and 30.3%, respectively, on postoperative day 1 following LC, and by 44.3%, 46.2% and 58.3%, respectively, after OC. Chest X-ray showed atelectasis in 15% of patients undergoing LC and 45% of those with OC. CONCLUSION: Impairment in pulmonary function after LC was less marked than after OC. PMID- 15778518 TI - Devadhar's operation for complete rectal prolapse: 25 years' experience. AB - BACKGROUND: Numerous abdominal and perineal operations have been described for the treatment of complete rectal prolapse. We describe our results with Devadhar's rectopexy, which avoids dissection in the presacral space and hence may be expected to have a low risk of sexual and urinary disturbances. METHODS: Case records of 72 consecutive patients (40 men), aged above 18 years, with complete rectal prolapse who were treated with Devadhar's operation were reviewed. RESULTS: The only complication observed was mucosal prolapse in 3 patients. None of the 40 men had erectile dysfunction or retrograde ejaculation after a median follow-up of 10 (range 3-48) months. No patient had disturbance in micturition. Two patients (2.7%) had recurrence of rectal prolapse. In four patients (8.5%), constipation persisted. CONCLUSION: Devadhar's rectopexy for complete rectal prolapse was not associated with disturbances in sexual or micturition function, and low rates of recurrence of prolapse. PMID- 15778519 TI - Role of HBV genotype in predicting response to lamivudine therapy in patients with chronic hepatitis B. AB - BACKGROUND: Predictors of response of chronic hepatitis B (CHB) to lamivudine therapy need better definition. Whether hepatitis B virus (HBV) genotypes could serve as such a predictor has not been well studied. AIM: To study the association of HBV genotypes with the outcome of lamivudine treatment in patients with CHB. METHODS: Seventy-six patients with CHB (45 HBeAg +ve) received lamivudine 100 mg/day, orally for 12 mo. Infecting HBV genotypes were determined in pre-treatment specimens using restriction fragment length polymorphism. End-of treatment response (ETR) and sustained viral response (SVR) were defined as undetectable HBV DNA (< 0.5 pg/mL) at 12 and 18 months, respectively. RESULTS: ETR was observed in 26 (34%) and SVR in 11 (14%) patients receiving lamivudine. The pre-treatment characteristics of the responders and non-responders were comparable. Genotypes A and D were observed in 28 (37%) and 48 (63%) patients, respectively. The frequency of genotypes A and D was comparable between responders (28.6% vs. 37.5%) and non-responders (71.4% vs. 62.5%), respectively (p=ns). Of the 26 responders, SVR could be evaluated in 20 subjects; 9 (45%) relapsed and 11 achieved SVR. Patients with genotype D achieved higher SVR rate than genotype A (10 of 48, 28.8% vs. 1 of 28, 3.5% p =0.0359). CONCLUSIONS: Forty five percent of Indian patients with CHB who achieve ETR relapse, and SVR to lamivudine therapy is achieved in 14%. Patients with genotype D achieve higher SVR rate than with genotype A. PMID- 15778520 TI - Pre-operative prediction of difficult laparoscopic cholecystectomy using clinical and ultrasonographic parameters. AB - BACKGROUND: Pre-operative prediction of a difficult laparoscopic cholecystectomy (LC) can help the patient as well as the surgeon prepare better for the intra operative risk and the risk of conversion to open cholecystectomy. METHODS: In 105 eligible patients who underwent LC during May 2001 to January 2003, patient characteristics, clinical history, laboratory data, ultrasonography results and intra-operative details were prospectively analyzed to determine predictors of difficult LC. RESULTS: Of 105 patients, 12 (11.4%) required conversion to open cholecystectomy. Significant predictors of conversion were body mass index> 30 Kg/m2, male gender, past history of acute cholecystitis or acute pancreatitis, past history of upper abdominal surgery, and gall bladder wall thickness exceeding 3 mm. CONCLUSION: Clinical and ultrasonograpic factors can help predict difficult LC and likelihood of conversion of LC to open surgery. PMID- 15778521 TI - Hepatitis B virus genotype A is more often associated with severe liver disease in northern India than is genotype D. AB - BACKGROUND: The clinical outcome of chronic hepatitis B may depend on hepatitis B virus (HBV) genotype. Data from India on this aspect are limited and contradictory. We studied the frequency of HBV genotypes and their clinical significance. METHODS: Stored sera from patients with chronic HBV infection were tested for HBV genotype using PCR-RFLP. Clinical data, and biochemical and serological parameters were retrieved from medical records; patients were classified as having chronic hepatitis or cirrhosis. RESULTS: Of 70 patients studied (mean age [SD] 38.4 [17.0] years; 63 men; ALT 140 [177] U/L), 32 had chronic hepatitis and 38 had cirrhosis. HBeAg was positive in 50/67 (75%), and anti-HBe in 12/66 (18%). Genotype A was the commonest (37; 53%), followed by D (32; 46%) and C (1; 1%). Patients with genotype A more often had ALT elevation exceeding 1.5 times normal (30/37 [81%] than those with genotype D (18/31 [58%]; p< 0.05). They also more often had positive HBeAg (32/37; 86%) and negative anti HBe (33/36; 92%) than those with genotype D (18/29 [62%] and 21/29 [72%], respectively; p< 0.05 each). Of 37 patients with genotype A, 23 (62%) had cirrhosis and 14 (38%) had chronic hepatitis; of 32 patients with genotype D, 15 (47%) had cirrhosis and 17 (53%) had chronic hepatitis (p=ns). In the subgroup aged> 25 years, genotype A patients more often had cirrhosis than those with genotype D (23/28 [82%] vs 13/23 [57%]; p < 0.05). CONCLUSION: HBV genotypes A and D were the commonest in our population. Genotype A was more often associated with ALT elevation, HBeAg positivity, absence of anti-HBe and, among those aged 25 years and above, cirrhosis of liver, than was genotype D. PMID- 15778522 TI - Increasing incidence of Crohn's disease in India: is it related to improved sanitation? PMID- 15778523 TI - Perforation of Meckel's diverticulum by roundworm. AB - An 11-month-old girl child presented with distension of abdomen, constipation, vomiting and fever. X-ray of the abdomen suggested intestinal obstruction. Exploration revealed the distal ileum to be kinked around a band running from the tip of a congested Meckel's diverticulum to the lateral pelvic wall. A small perforation was noticed at the tip of the diverticulum and a live roundworm was found lying free in the peritoneal cavity. The diverticulum along with a segment of adjoining bowel was resected and end-to-end anastomosis done. The diverticulum had a wide lumen and showed no heterotopic tissue on subsequent examination. PMID- 15778524 TI - Hepatic tuberculosis mimicking carcinoma gall bladder. AB - We report a 50-year-old man with tuberculosis of the liver with calculous cholecystitis. The diagnosis was made when the patient underwent cholecystectomy. He responded to antitubercular therapy. PMID- 15778525 TI - Gigantic post-traumatic pseudocyst of sigmoid mesentery. AB - A 29-year-old man presented with dull abdominal pain and a lump occupying almost the entire abdomen, four months after a fall from a height. Ultrasonography revealed a cystic lesion with debris occupying almost the entire abdomen. Diagnostic tap revealed brownish fluid. Exploration revealed a huge thick-walled cyst of the sigmoid mesocolon, which could be enucleated out entirely. Histology suggested it to be a false cyst. PMID- 15778526 TI - Bifid liver in a patient with diaphragmatic hernia. AB - Liver malformations including lobe and segmental abnormalities are rare. We report a 65-year-old lady with complaints of breathlessness and fullness after meals for two months. Investigations revealed a diaphragmatic hernia on the right side with a bifid liver; the right lobe of the liver was among the hernia contents. The lady is asymptomatic after surgical repair. PMID- 15778527 TI - Pleuro-biliary fistula--a delayed complication following open cholecystectomy. AB - A 24-year-old lady presented with bilious expectoration and history suggestive of obstructive jaundice, 16 months following open cholecystectomy. Pleurocentesis was done, followed by intercostal drainage, which drained about 300 mL bilious fluid per day for a month. Her symptoms of obstructive jaundice were relieved. A pleuro-biliary fistula was demonstrated by percutaneous transhepatic cholangiogram, HIDA scan and ERCP, which showed complete cut-off at the lower end. The patient underwent bilio-enteric bypass with gastric access loop, with complete healing of the pleuro-biliary fistula. PMID- 15778528 TI - Hepatic resection for metastasis from adenoid cystic carcinoma of parotid gland. AB - Adenoid cystic carcinoma is the commonest malignant tumor of the submandibular and minor salivary glands; the parotid gland constitutes a small share of this neoplasm. We present a 30-year-old woman with solitary liver metastasis from an adenoid cystic carcinoma of the parotid gland, which had been surgically treated 10 years ago. The patient underwent successful resection of this metastasis. PMID- 15778529 TI - Atypical Whipple's disease. AB - We report an unusual case of Whipple's disease diagnosed with help from the patient himself. The patient presented with rash resembling dermatitis herpetiformis, constipation, and intermittent diarrhea. A suspicion of celiac disease led to duodenal biopsy, which showed features of Whipple's disease on histology and electron microscopy. PMID- 15778530 TI - Role of fruit consumption in gall bladder cancer. PMID- 15778531 TI - HLA class I and II antigens associated with susceptibility and resistance to ulcerative colitis. PMID- 15778532 TI - Prevalence of hepatitis A virus antibodies in patients with chronic liver disease in Shiraz, Iran. PMID- 15778533 TI - Technical modification for difficult intubation during endoscopic variceal ligation. PMID- 15778534 TI - Hepatitis A in pediatric acute liver failure in southern India. PMID- 15778536 TI - Autoimmune pancreatitis. PMID- 15778537 TI - Dysphagia due to olanzepine, an antipsychotic medication. PMID- 15778538 TI - Sporadic, multiple adenomatous polyposis coli treated with ileo-endorectal pull through. PMID- 15778539 TI - Esophageal bezoar in a patient with normal esophagus. PMID- 15778540 TI - [CMV colitis in diabetic patient without use of immunosuppressant]. PMID- 15778541 TI - [Photodynamic therapy in gastroenterology]. AB - Photodynamic therapy (PDT) was first used for the treatment of esophageal cancer in early 1980s, Since then, numerous applications have been reported for its use in gastrointestinal tract including Barrett's esophagus, gastric, duodenal, biliary, pancreatic and colorectal lesions. PDT in gastroenterology has made tremendous progress over the last decade but its clear role is yet to be proved. Now, there is an increasing need for less invasive methods of treatment in patients with pre-malignant disease, early cancer or those who are unfit for surgery. It is one of a number of ablative techniques currently under investigation and appears to have a number of potential advantages over other forms of treatment in the alimentary tract. The development of newer potent, highly efficient photosensitizers, as well as endoscopic imaging techniques and light delivery systems, are continuing to expand the clinical uses of PDT. As data from additional clinical trials become available, we will gain a clearer perspective of where PDT fits in the treatment of cancers. PMID- 15778542 TI - [Frequency analysis of NOD2 gene mutations in Korean patients with Crohn's disease]. AB - BACKGROUND/AIMS: Several studies from Western populations have recently shown that three mutations in NOD2 gene (C2104T, G2722C, and 3020insC) are associated with susceptibility to Crohn's disease (CD). However, three mutations were shown not to be associated with CD in Japanese and Chinese population. Here, we have analyzed the frequency of three NOD2 mutations in Korean patients to determine whether the NOD2 mutations are associated with susceptibility to CD in Korean population. METHODS: Blood samples were obtained from 128 patients with CD, 47 patients with ulcerative colitis, 19 Behcet's colitis, and 200 healthy controls. DNA in the region of three NOD2 mutations was sequenced by single base extension method, and the frequency of mutations were analyzed. RESULTS: Among the subjects in our study groups, including patients with CD, ulcerative colitis, Behcet's colitis, and healthy controls, none had NOD2 mutations. CONCLUSIONS: Our results indicate that although three NOD2 mutations are associated with susceptibility to CD in Western populations, these might be rare and may not be associated with susceptibility to CD in Korean patients. PMID- 15778543 TI - [Clinical analysis of prognosis in spontaneous esophageal rupture]. AB - BACKGROUND/AIMS: Spontaneous esophageal rupture is a life-threatening injury because of delay in diagnosis and rapid progression to septic condition. But acceptable standard treatment strategy has not been established yet. This may be due to its low incidence and lack of published literature. In this study, we evaluated the proper treatment strategy as to decide when and how to manage spontaneous esophageal rupture by analyzing our experiences. METHODS: Eleven patients who were diagnosed as spontaneous esophageal perforation in Dongsan Medical Center from 1993 to 2003 were analyzed. Patients WERE divided into survival and death group. Clinical manifestations, rupture site and size, treatment methods and complications were compared. RESULTS: All patients were male and alcoholics. Six patients had survived and five patients died. Age, sites and sizes of ruptures, operation methods were not different in both groups. Before operation, septic condition was present in all patients of death group and more common than survival group (p=0.015). Survival group showed shorter time interval from rupture to initial treatment (p=0.021) and to operation (p=0.019). CONCLUSIONS: Early diagnosis and initial aggressive treatments such as nothing per oral, pleural and mediastinal drainage are important factor for better prognosis. If possible, early operation such as primary repair must be done. Although any difference according to types of operation method was not found in this study, further study with larger groups seems to be necessary. PMID- 15778544 TI - [Therapeutic effect of adefovir dipivoxil on recurrent or de novo infection of hepatitis B virus after liver transplantation: a preliminary report]. AB - BACKGROUND/AIMS: Anti-viral therapy using hepatitis B immune globulin and lamivudine could not prevent HBV recurrence after liver transplantation (LT) completely. Adefovir dipivoxil is a acyclic nucleotide phosphate analogue and known to have potent anti-HBV effect. In this study, we analyzed the therapeutic effect of adefovir for recurrent or de novo HBV infection after LT. METHODS: From December 2002 to October 2004, adefovir was administered in 12 post-LT patients of HBV infection (11 recurrent and 1 de novo infection). In these patients, lamivudine and other combined therapies were used before the introduction of adefovir. Thereafter, adefovir combined with lamivudine was administered to all patients. RESULTS: The duration of adefovir administration was 5.5-18 (median, 15.5) months. The median values of serum AST and ALT levels were significantly reduced from 86+/-80 IU/L and 140+/-103 IU/L, respectively before the adefovir administration to 42+/-19 IU/L and 38+/-33 IU/L after 2 months of administration. This trend of improved liver function persisted throughout the follow-up period. HBeAg seroconversion was achieved in 4 of 10 patients (40%) and HBsAg seroconversion was observed in 1 of 10 patients (10%). HBV DNA levels have decreased to undetectable levels by hybridization assay in 6 of 7 patients within the first 2 months of therapy. Nephrotoxicity and hypophosphatemia were not found in all of these patients. CONCLUSIONS: Based on this preliminary result, adefovir dipivoxil seems to be an effective and safe antiviral agent leading to viral inhibition and clinical improvement in post-LT patients with recurrent or de novo HBV infection. PMID- 15778545 TI - [Positive predictability and predictive factors of the third generation anti hepatitis C virus (HCV) ELISA test for HCV infection]. AB - BACKGROUND/AIMS: Anti-HCV positivity suggests past or present infection of HCV, or false positivity. The positive predictability of this test can differ according to the subjects. This study examines the positive predictability of the third generation anti-HCV ELISA and factors predicting HCV infection with special emphasis on the significance of the anti-HCV sample/cut-off (S/CO) ratio. METHODS: One hundred and ninety patients who were anti-HCV positive were enrolled, from November 1998 to January 2002 in Kyung Hee University Hospital. RT PCR was performed to confirm HCV infection. RESULTS: One hundred and seven patients were RT-PCR positive (56.3% positive predictability). The positive predictability changed with the S/CO ratio: 17.9% in cases below 6, 58.3% between 6 and 50, 78.6% between 51 and 75, and 60% over 75. Those with the S/CO ratio more than 6 showed significantly higher predictability, but it did not increase further when the ratio got higher. Factors predicting HCV infection were the presence of liver cirrhosis (OR 5.5, p=0.000), hepatocellular carcinoma (OR 11.67, p=0.004), liver diseases (OR 2.99 p=0.001), and increase of AST (OR 2.49, p=0.002), ALT (OR 2.32, p=0.005), alpha-FP (OR 3.49, p=0.040), and the S/CO ratio of more than 6 (OR 7.82, p=0.000). However, liver cirrhosis was the sole factor in multivariate analysis (OR 8.32, p=0.02). CONCLUSIONS: The positive predictability of the third generation anti-HCV test was 56.3% with a significant difference between those with the S/CO ratio below 6 (18%) and above 6 (63%). In liver cirrhosis, positive predictability of anti-HCV test was relatively high as 85%. PMID- 15778546 TI - [Immunohistochemical study of p53 mutation and p16, p14 alterations encoded by INK4a-ARF in mucin-hypersecreting bile duct tumor]. AB - BACKGROUND/AIMS: Mucin-hypersecreting bile duct tumor is rare, and has an unusual histologic characteristic of having various degrees of cellular atypia ranging from dysplasia to invasive carcinoma in the same specimen. To gain insight into the role of p16, p14 and p53 in the carcinogenic process of bile duct tumor, we analyzed the expression status of these proteins in mucin-hypersecreting bile duct tumor. METHODS: Immunohistochemical staining of p16, p14 and p53 were performed in 34 paraffin embedded tissues obtained from 22 patients of mucin hypersecreting bile duct tumor. RESULTS: Thirty-four specimens were categorized into low-grade dysplasia (9), high-grade dysplasia (4), carcinoma in situ (CIS, 11) and invasive carcinoma (10) based on the degree of cytologic and structural atypia. p53 overexpressions were found in 6 (17.6%, 3 in CIS, 3 in invasive carcinoma) and more frequently observed in the advanced histologic stages (p<0.05). Loss of p16 staining was found only in 2 (6%) of low-grade dysplasia specimen. Loss of p14 staining was found in 21 (61.7%, 7 in low-grade dysplasia, 2 in high-grade dysplasia, 8 in CIS, and 4 in invasive carcinoma) and was frequently observed in low-grade and high-grade dysplasia compared to p53 (p<0.05). CONCLUSIONS: In mucin-hypersecreting bile duct tumor, p14 and p53 may play a role in the early and advanced stage of carcinogenesis, respectively. Further study regarding genetic and epigenetic alterations in p14 and p53 gene may be needed. PMID- 15778547 TI - [Two cases of metronidazole-induced encephalopathy]. AB - Metronidazole is a 5-nitroimidazole compound known as an antimicrobial agent widely used for the treatment of protozoal infection, anaerobic infection, Helicobacter pylori infection and hepatic encephalopathy. It may produce a number of neurologic side effects including peripheral neuropathy, seizure, encephalopathy, ataxic gait and dysarthritic speech. There have been ten or more reports of metronidazole-induced encephalopathy in the literatures including a few reports of brain imaging changes by magnetic resonance images (MRI). However, none of the case of metronidazole-induced encephalopathy in patients with hepatic encephalopathy has been reported yet. Recently, we experienced two cases of metronidazole-induced encephalopathy in patients with liver cirrhosis caused by chronic hepatitis B, which were diagnosed by brain MRI and MR spectroscopy. In this report, we present 2 cases of metronidazole-induced encephalopathy with MR imaging and MR spectroscopic changes including follow-up imaging performed after the discontinuation of the metronidazole with a review of the literatures. PMID- 15778548 TI - [A case of Budd-Chiari syndrome and superior mesenteric vein thrombosis in ulcerative colitis]. AB - Patients with inflammatory bowel disease are at increased risk for thromboembolic complications. Its prevalence rate ranges from 1% to 7% in clinical studies and reaches 39% in autopsy series. The cause of thrombotic complications in inflammatory bowel disease is generally considered to be associated with hypercoagulability. We experienced a case of ulcerative colitis associated with Budd-Chiari syndrome and superior mesenteric vein thrombosis. This rare complication of ulcerative colitis was successfully managed by conventional treatment for ulcerative colitis and anticoagulation therapy. PMID- 15778549 TI - [A case of colon obstruction developed during the recovery period of acute pancreatitis]. AB - Complications of acute pancreatitis usually occur in pancreas and its contiguous organs. The prevalence of colonic invasion is rare, however, the consequence is fatal, with mortality above 50%. The initial symptoms and onset times are variable and major affected sites are transverse colon and splenic flexure. The spread of inflammatory exudates into the colon is the main mechanism of colonic invasion. If the colonic stenosis develops, it is necessary to manage it surgically. We report a case who arrived at the hospital with watery diarrhea and abdominal distension in the recovery period of acute alcoholic pancreatitis and was diagnosed as a colonic obstruction in the splenic flexure. The patient underwent loop ileostomy instead of the resection of the lesion because of severe adhesion around the splenic flexure. The patient died due to sepsis 5 days after the operation. PMID- 15778550 TI - [Effect of Helicobacter pylori infection on the cardiovascular diseases]. PMID- 15778552 TI - [Development of an interaction behaviors checklist for early detection of autistic children]. AB - PURPOSE: This study was conducted to develop a behavioral checklist to predict an autistic disorder and to identify the earliest detecting time. METHOD: One hundred and fifty eight children including normal, autistic, institutionalized normal, and retarded were assessed using critical interaction behavioral markers from literature review. Data was collected by semi-structured mother-child interaction by videotape recording and analyzed by factor analysis, Cronbach alpha, Kappa, chi(2), and Duncan. RESULT: Ten behavioral markers were sorted into 2 factors; joint-attention and synchronized behavior. Autistic children were impaired in pretend play, prodeclarative pointing, proimperative pointing, gaze monitoring, referential looking, showing, joint-attention, rhythmical vocal exchange, and synchronized laughing. The sychronized behavior was also a critical marker to predict the autistic disorder. However, it was difficult to differentiate autistic disorder from mental retardation. In addition, the appropriate detecting time was around 18 months after birth. CONCLUSION: This checklist should be behavior markers to predict autistic disorder and could be useful as educational material at children's clinics, parents class, and for caregivers in the health center. In addition, early detection should lead to treatment being started as soon after 18 months of age as possible. PMID- 15778553 TI - [Psychosocial adjustment process in adolescents with epilepsy]. AB - PURPOSE: The purpose of this study was to explore the psychosocial adjustment process in adolescents with epilepsy in the context of Korean society and culture. METHOD: A grounded theory method was used for data collection and analysis. Participants for this study were 9 adolescents who regularly visited an epilepsy clinic in a university hospital. The data was collected through in-depth interviews during the period from November, 2002 to June, 2003. Data collection and analysis were performed simultaneously. RESULT: Twenty-three categories emerged including 'suffering', 'psychological stigma', and 'social isolation from one's peers'. Categories were divided into paradigms which consisted of conditions, actions/interactions, and consequences. 'Reconstructing life' was the core category in this study. The theoretical scheme was described by organizing categories around the core category. CONCLUSION: This study provides a framework for the development of individualized nursing interventions to care for adolescents with epilepsy. PMID- 15778554 TI - [Prospective study on the relating factors to the stages of change in smoking cessation and barriers in coronary artery disease patients]. AB - PURPOSE: The main purpose of this study was to investigate that the stages of change in smoking cessation behavior among coronary artery disease patients for six months progressed following the stages of change suggested by the transtheoretical model. METHOD: Subjects for this descriptive survey were 59 coronary disease patients who were smoking or who had stopped smoking for less than six months. RESULT: In the baseline, the distribution of the subjects' stages of change was as follows: pre-contemplation stage 25.4%, contemplation stage 25.4%, preparation stage 22%, and action stage 27.1%. After six months, more subjects in the contemplation(33.3%) and preparation stages(30.8%) progressed to the action stage than those of the pre-contemplation stage(0%). Eighty-one percent of the subjects in the action stage at baseline progressed to the maintenance stage. The relationship between the numbers of smoking cessation attempts for six months and stages of change at baseline was significant(p=.001). However, the relationships between self-efficacy and nicotine dependence at baseline and progression in stages of change after six months were not significant. CONCLUSION: Progression in the stages of change for six months among subjects corresponded to the stages of change suggested by the transtheoretical model. Hence, future development and evaluation of intervention programs should be tailored individually considering each patient's stage of change. PMID- 15778555 TI - [Canonical correlation between Korean traditional postpartum care performance and postpartum health status]. AB - PURPOSE: The purpose of this longitudinal study was to examine the relationship between postpartum care performance and postpartum health status. SAMPLE: The study subjects were 82 mothers who delivered full-term infants at 3 hospitals at P city. Data were collected for their health status at the postpartum unit and the sample was followed up to 6 weeks postpartum to collect postpartum care performance and health status. RESULTS: Mothers rated postpartum care performance as moderate to high and especially rated the maternal role attainment the highest. Mothers experienced 4 physical symptoms and moderate levels of fatigue. In addition, they experienced moderate levels of positive affect and low levels of negative affect at both times. Canonical correlation revealed that postpartum care performance was related to postpartum health status with 2 significant canonical variables. The first variate indicated that mothers who performed hospitality, physical and emotional recovery, self-caring, and role attainment well showed higher positive affects, lower negative affects, fewer physical symptoms, and lower levels of fatigue. The second variate showed that the greater the performance of caring and physical and emotional recovery, the fewer physical symptoms and lower levels of fatigue. CONCLUSION: Although Korean traditional postpartum care performance was related to postpartum health status, the further study is needed to identify the causal relationship between them. Nurses need to integrate the perspective of westernized postpartum care and Korean traditional views of postpartum approach to maintain and promote women's health better. PMID- 15778556 TI - [Evaluating the validity of the Pediatric Index of Mortality II in the intensive care units]. AB - PURPOSE: This study was to evaluate the validity of the Pediatric Index of Mortality II(PIM II). METHOD: The first values on PIM II variables following ICU admission were collected from the patient's charts of 548 admissions retrospectively in three ICUs(medical, surgical, and neurosurgical) at P University Hospital and a cardiac ICU at D University Hospital in Busan from January 1, 2002 to December 31, 2003. Data was analyzed with the SPSSWIN 10.0 program for the descriptive statistics, correlation coefficient, standardized mortality ratio(SMR), validity index(sensitivity, specificity, positive predictive value, negative predictive value), and AUC of ROC curve. RESULT: The mortality rate was 10.9%(60 cases) and the predicted death rate was 9.5%. The correlation coefficient(r) between observed and expected death rates was .929(p<.01) and SMR was 1.15. Se, Sp, pPv, nPv, and the correct classification rate were .80, .96, .70, .98, and 94.0% respectively. In addition, areas under the curve(AUC) of the receiver operating characteristic(ROC) was 0.954(95% CI=0.919-0.989). According to demographic characteristics, mortality was underestimated in the medical group and overestimated in the surgical group. In addition, the AUCs of ROC curve were generally high in all subgroups. CONCLUSION: The PIM II showed a good, so it can be utilized for the subject hospital. PMID- 15778557 TI - [Effect of aromatherapy massage for the relief of constipation in the elderly]. AB - PURPOSE: The purpose of this study was to verify the effect of aromatherapy massage on constipation in the elderly. METHOD: This study for 10 day, employed a randomized control group pretest-posttest design. The experimental group received abdominal massage using essential oils with Rosemary, Lemon, and Peppermint, and the control group received a placebo massage. To evaluate the effect of aromatherapy, the degree of constipation was measured using the CAS(constipation assessment scale) and the number of bowel movements per week. Data was analyzed by repeated measures of ANOVA using the SPSS program. RESULT: The score of CAS of the experimental group was significantly lower than that of the control group. In addition the average number of bowel movements in the experimental group was higher than that of the control group. The effect of aromatherapy lasted 2 weeks after treatment, while the placebo effect lasted 7-10 days after treatment. CONCLUSION: The finding of this study showed that aromatherapy helps relieve constipation in the elderly. PMID- 15778558 TI - [Perception about problem-based learning in reflective journals among undergraduate nursing students]. AB - OBJECTIVE: The aim of this study is to explore the variation in perceptions about problem-based learning(PBL) according to the level of academic achievement and learning attitude in the nursing students of a junior college (3-year program). METHOD: Students (n=39) learned the respiratory and cardiac system with seven PBL packages and group-based learning for a semester in 2002. Students were asked to write reflective journals that focused on their learning perception after an experience with each learning package. A total of 208 journals were used for analysis. RESULT: Students positively perceived that PBL making them increase their sense of responsibility for learning and felt satisfaction with the learning process, and had a confidence in the use of clinical nursing interventions. On the other hand, they negatively perceived that PBL was a burden because it took more time than traditional learning tasks, and they experienced an anxiety about regular tests and felt conflicts and diffidences in the learning process. The negative perceptions were expressed more often from students with a low academic achievement and low learning attitude compared to others. CONCLUSION: Students perceived the PBL as effective in understanding the learning concepts in the clinical practice environment. PBL need to be supplemented by feedback-based lecture and facilitative strategies for academically low-achieved students. PMID- 15778559 TI - [The effects of a stress management program on mental health and the coping behavior for the children of alcoholics]. AB - PURPOSE: The purpose of this study was to examine the effects of a stress management program on mental health and coping behavior for children of alcoholics. METHOD: Data was collected from January to February, 2003. The subjects were 20 adolescents from 13 to 18 years old. Data was analyzed using descriptive statistics, chi-square test, and t-test with the SAS program. RESULT: There were statistically significant differences in mental health, active coping, positive cognitive restructuring, and support-seeking for problem solving between the experimental group and the control group. CONCLUSION: The stress management program helped children of alcoholics by enhancing self-esteem, providing information about alcohol, and improving emotional and problem focused coping abilities. This eventually enhanced mental health. PMID- 15778560 TI - [Effecting factors on depression in patients with fibromyalgia]. AB - PURPOSE: The purpose of this research was to provide basic data in the control of depression found in patients with fibromyalgia by analysing the main factors that affect them. METHOD: The subjects were 207 out-patients in three university medical centers diagnosed with fibromyalgia according to the conditions by American College of Rheumatology (1990). Depression was measured by the CES-D Scale. Dependant variables, such as stress and anxiety were measured by the VAS Scale and physical activity was measured using 10(4-point-scale) questions developed by the researcher. The number of tender points was converted to scores based on the criteria of the ACR(1990) and of Yunus. Self-efficacy was measured by the Self-Efficacy Scale developed by Lorig et al.(1989) for arthritis patients. RESULT: The main factors that affect depression of patients with fibromyalgia were self-efficacy, anxiety, physical activity, tender points fatigue, and family support, which explained 49% of the depression. CONCLUSION: It has been confirmed that the regression equation model of this research may serve as a predictor of depression in patients with fibromyalgia. PMID- 15778561 TI - [Development of a task performance evaluation instrument for clinical nurses]. AB - PURPOSE: The purpose of this study was to develop an instrument of task performance evaluation for clinical nurses, thus testing the validity and the reliability of the scale. METHOD: Data was collected from 84 Head Nurses and 255 General Nurses. A conceptual framework, composed of 4 factors of meaning in task performance evaluation, was identified through review of the relevant literature. A total of 78 items were developed and were used on a five-point likert scale. Through factor analysis, items whose factor loading was below 0.50 were deleted, thus 35 items remained. To test the validity and reliability of the instrument, the SPSS 11.0 windows program was used. RESULT: The results of the factor analysis indicated that 4 factors were classified and the cumulative percent of variance was 67.54%. The results of the reliability test indicated that Cronbach's coefficient of the total 35 items was over 0.9176. The results of the factor analysis indicated that factor loadings of all items was over 0.50. Conclusively, the validity and the reliability of the scale were proven. CONCLUSION: This study was identified as a tool with a high degree of reliability and validity. PMID- 15778562 TI - [The effects of fall prevention program on knowledge, self-efficacy, and preventive activity related to fall, and depression of low-income elderly women]. AB - PURPOSE: This research was conducted to determine the effects of a fall prevention program on knowledge, self-efficacy, prevention activity, and depression in the low-income elderly women. METHOD: The design of this study was a nonequivalent control group pretest-posttest design. There were 22 subjects in the experimental group and 22 in the control group. A fall prevention program was performed for approximately 40 minutes, once a week for 6 weeks. RESULTS: The results of this study were as follows: There were statistically significant differences of knowledge, self-efficacy, and prevention activity related to falls and depression between the experimental group and control group. CONCLUSION: In conclusion, the fall prevention program used in this study is appropriate for the elderly, therefore this program is strongly recommended for community-based health and welfare centers. PMID- 15778563 TI - [Scale development on health conservation of the institutionalized elderly]. AB - PURPOSE: The purpose of this study was to develop a health conservation scale with high validity and reliability for institutionalized elderly. METHOD: The process of development of this scale was as follows. A conceptual framework composed of 4 phases of health conservation of institutionalized elderly was identified based on the literature review with elderlies and discussions with experts in health conservation. A total of 75 items, on a 4-point scale were developed. Through reliability testing and factor analysis, 57 preliminary items were selected. By means of internal consistency of the 57 items, 18 items whose inner-items correlation coefficient was below .40 were deleted. Through factor analysis, 2 items whose factor loading was below .40 were deleted. Finally 37 items remained. To verify the 37 items, factor analysis, reliability testing, and correlation was done. Data were collected from 207 institutionalized elderly subjects in Daegu, Kyungpook, Busan, and KyungNam Province from August. 2003 to February. 2004. RESULT: In the result of factor analysis of the 37 items, 4 factors were extracted. These factors were labeled as 'personal integrity', 'conservation of energy', 'structural integrity', and 'social integrity'. These factors included 4 phases of health conservation. Cronbach's Alpha of 37 the items was .9424 and the correlation coefficient of HPLP was .723. CONCLUSION: The researchers recommend the following: An explorative study on the variables related to health conservation is needed for criterion validity of this scale. Studies on health conservation of different age groups, and subjects are needed for verification. PMID- 15778564 TI - [Factors influencing the development of pressure ulcers in surgical patients]. AB - PURPOSE: The purpose of this study was to identify the influencing factors on the development of pressure ulcers in patients undergoing surgery which lasted more than two hours. METHOD: One hundred nineteen surgical adult patients were included in the study. Data was measured on each participant from December 2003 to February 2004. It was collected using a structured researcher-administered sheet and analyzed by descriptive statistics, t-test, chi-square test and logistic regression analysis. RESULT: The prevalence of a perioperative pressure ulcer was 26.1%. The level of moisture, friction and shear, length of surgery, and perioperative irrigation were significantly higher in the pressure ulcer group than those in the non-pressure ulcer group. The level of activity and level of consciousness were significantly lower in the pressure ulcer group than those in the non-pressure ulcer group. Significant influencing factors on the development of pressure ulcer were 'moisture' and 'irrigation' and those variables explained 23.1% of variance in the development of a pressure ulcer during surgery. CONCLUSION: It is necessary to develop a strategy to prevent pressure ulcer by taking 'moisture' and 'irrigation' into account during the preoperative, perioperative and postoperative period. PMID- 15778565 TI - [The effects of abdominal meridian massage on constipation among CVA patients]. AB - PURPOSE: This study was aimed at developing an efficient nursing method for the management of constipation by comparing the effects of abdominal meridian massage on the symptoms of constipation. METHOD: The subjects were determined by the Rome II criteria and the Constipation Assessment Scale from Aug 30 through Sep 26, 2002. They were randomly divided into two groups: one experimental group of 16, another control group of 15. Abdominal meridian massage was given to the experimental group and no massage was given to the control group. The frequency of defecation and severity of constipation by CAS were measured. The data was analyzed with the t-test, chi(2)-test, and repeated measures ANOVA. RESULTS: There was a significant improvement in frequency of defecation, and severity of constipation in the experimental group compared to the control group. CONCLUSION: Abdominal meridian massage can be considered an effective nursing method for the management of constipation among CVA patients. PMID- 15778566 TI - [Effects of cycled lighting on body weight, physiological variables and behavioral states in low birth weight infants]. AB - PURPOSE: This study was aimed at finding the effects of cycled lighting on body weight, physiological variables and the behavioral state of LBWI (low birth weight infants) in the NICU. METHOD: The subjects were 20 LBWI at 2 NICUs. They were assigned to an experimental or control group which consisted of 10 subjects in each. Cycled light was applied to the experimental group for 10 days. RESULT: It was certified that the application of cycled lighting resulted in increased body weight and O(2) saturation, and decreased heart rate of the LBWI. However, there was no effect in decrease of respiration and stabilization of the behavioral state. CONCLUSION: The application of cycled lighting might be a nursing intervention which would in turn have positive effects on the growth of LBWI. PMID- 15778567 TI - [Effects of an empowerment program on the burden of mothers having a child with cerebral palsy]. AB - PURPOSE: The purpose of this study was to develop an empowerment program as a nursing intervention for mothers who care for a child with cerebral palsy at home and to determine the effects of the program on those mothers' self efficacy, coping behavior and burden. METHOD: An non-equivalent control group pretest posttest design was used in this study. An Empowerment program was developed based on Dunst & Trivette's model. Using the program, the study was carried out from Dec. 13, 2003 to Jan. 17, 2004, mothers whose children, aged 1 to 6, were outpatients of the Dept. of Rehabilitation Medicine, at P University Hospital or registered at educational institutions for early disabled children. The experimental group of subjects were included in the new empowerment program which was held for two and half hours every week for 6 times. RESULTS: After treatment with the Empowerment Program, the experimental group was found to be significantly increased in score for self efficacy(t=4.55, p<.01), coping behavior(t=5.54, p<.001), objective burden(t=-3.96, p<.01) and subjective burden(t=-5.05, p<.01), in comparison to the control group. CONCLUSION: The Empowerment Program is very effective in increasing self efficacy and coping behavior of mothers having a child with cerebral palsy and decreasing their burden. Finally, this study would recommend that an empowerment program should be extended to community facilities such as public health offices and welfare centers. PMID- 15778568 TI - [Development and evaluation of a Web-based support program for the maternal role of primiparas]. AB - PURPOSE: This study was conducted to develop and evaluate a Web-based program for the maternal role of primiparas who use the internet. METHOD: The study process was a systems requirements analysis, design and development of a program, program testing by experts, program implementation, and program evaluation by users. A nonequivalent control group non-synchronized design was used. The data was collected from October 5th, 2002 to February 24th, 2003. RESULT: 1. Based on inquiries into mothers' needs, a Web-based support program was developed. The program was then modified from feedback received from experts. 2. In a sub-scale analysis of mothers' perception of a baby, amenability and persistence was significantly higher in the intervention group. The differences in the mean score of maternal self-confidence and maternal satisfaction were significant. 3. Cyber counseling was done for a total of 73 cases and the most frequent problems for counseling were feeding and nutrition (28.8%), followed by baby care, and health problems. CONCLUSION: It was proven that a Web-based support program provided appropriate support to primiparas and was effective in promoting their maternal role. Therefore, this study suggests that a Web-based support program for primiparas can become a powerful nursing intervention on virtually all mother and infant health concerns. PMID- 15778569 TI - [The effects of foot reflexology on nausea, vomiting and fatigue of breast cancer patients undergoing chemotherapy]. AB - PURPOSE: The purpose of this study was to identify the effects of foot reflexology on nausea, vomiting and fatigue in breast cancer patients undergoing chemotherapy. METHOD: The research was a quasi-experimental study using a non equivalent pre-post design and was conducted from Jan. 26, to Mar. 20, 2004. The subjects consisted of 34 patients with 18 in the experimental group and 16 in control group. A pretest and 2 posttests were conducted to measure nausea, vomiting and fatigue. For the experimental group, foot reflexology, which was consisted of 4 phases for 40 minutes, was given by a researcher and 4 research assistants. The collected data were analyzed by repeated measures ANOVA using the SPSS WIN 10.0 program. RESULTS: There was a statistically significant decrease in nausea, and vomiting in the experimental group compared to the control group over two different times. In addition, there was a statistically significant decrease in fatigue in the experimental group compared to the control group over two different times. CONCLUSION: Foot reflexology was effective on nausea, vomiting and fatigue in breast cancer patients receiving chemotherapy in this study. Therefore, foot reflexology can be usefully utilized as a nursing intervention in the field of cancer nursing for breast cancer patients receiving chemotherapy. PMID- 15778571 TI - [A correlational study among perceived stress, anger expression, and depression in cancer patients]. AB - PURPOSE: This study was to identify the relationship between perceived stress, anger expression, and level of depression in cancer patients. METHOD: A cross sectional descriptive study design was used. Data was collected by questionnaires from 185 in- and out-patients who were diagnosed with cancer at 3 university hospitals and the National Cancer Center using Spielberger et al.'s Anger Expression Scale, Cohen, Kamarch & Mermelstein's Perceived Stress, and Derogatise's SCL-90. The data was analyzed using descriptive statistics, Pearson correlation coefficient, and stepwise multiple regression with SAS/PC. RESULT: The perceived stress in cancer patients indicated a significant positive correlation to anger-in(r=.288, p=.000), anger-out(r=.232, p=.001), and depression(r=.68, p=.000), but no significant correlation to anger-control. The anger-in of cancer patients showed a significant positive relationship to anger out(r=.53, p=.000), and depression(r=.383, p=.000), but no significant correlation to anger-control. Anger-out showed a significantly negative correlation to anger-control(r=-.248, p=.001) and a positive correlation to depression(r=.240, p=.001). The most significant predictor which influenced depression in cancer patients was perceived stress, followed by anger-in and hobby, and these factors explained their depression with a variance of 54%. CONCLUSION: These results suggested that cancer patients with a high degree of perceived stress are likely to be high in anger-out and anger-in. Perceived stress and anger-in are major factors which affect depression in cancer patients. PMID- 15778570 TI - [The effects of aromatherapy on pain, depression, and life satisfaction of arthritis patients]. AB - PURPOSE: The purpose of this study was to investigate the effect of aromatherapy on pain, depression, and feelings of satisfaction in life of arthritis patients. METHOD: This study used a quasi-experimental design with a non-equivalent control group, pre-and post-test. The sample consisted of 40 patients, enrolled in the Rheumatics Center, Kangnam St. Mary's Hospital, South Korea. The essential oils used were lavender, marjoram, eucalyptus, rosemary, and peppermint blended in proportions of 2:1:2:1:1. They were mixed with a carrier oil composed of almond (45%), apricot(45%), and jojoba oil(10%) and they were diluted to 1.5% after blending. The data were analyzed using an 2-test, Fisher's exact test, t-test and paired t-test. RESULT: Aromatherapy significantly decreased both the pain score and the depression score of the experimental group compared with the control group. However, aromatherapy didn't increase the feeling of satisfaction in life of the experimental group compared with the control group. CONCLUSION: The result of this study clearly shows that aromatherapy has major effects on decreasing pain and depression levels. Based on our experiment's findings, we suggest that aromatherapy can be a useful nursing intervention for arthritis patients. PMID- 15778572 TI - [Analysis on articles published in Journal of Korean Academy of Nursing (Volume 33, Number 6-Volume 34, Number 5) based on nursing knowledge classifications]. PMID- 15778573 TI - Population biokinetic modeling of thyroid uptake and retention of radioiodine. AB - The aim of this study was to build a population biokinetic model for the uptake and retention of radioiodine in the thyroid of hyperthyroid patients. At Malmo University Hospital (Malmo, Sweden), a database with detailed information from (131)I-iodide therapies has been created. The database consisted of uptake measurement from 422 patients with the diagnoses of diffuse goiter, multinodular goiter, adenoma, and undefined. In total, 2013 uptake measurements, performed from 3 hours to 9 days after intake of a test activity of (131)I, were included. Nonlinear mixed effect modeling (NLME) was used to model the population biokinetics of (131)I. The structural model was parameterized in terms of the level of uptake (A), the rate constant for the absorption to the thyroid (k(a)) and the rate constant for the output from the thyroid (k(e)). Interindividual and interoccasion variabilities were added to all three structural parameters. The following covariates were found to significantly affect the structural model parameters: Diagnosis on A, k(a) and K(e), volume of the thyroid on A and k(a), and age on k(a) and k(e). The interindividual and interoccasion variabilities were in the range of 8%-28%. The variance of the residual error, modeled as an additive, was low. This type of modeling is a powerful tool for biokinetic studies and should be used in other biokinetic areas of radioiodine therapy, such as radiation dosimetry. PMID- 15778574 TI - Administration of high-dose continuous infusion interleukin-2 to patients age 70 or over. AB - High-dose bolus or continuous infusion interleukin-2-based therapy can cause capillary leak syndrome. Significant cardiovascular/hemodynamic events, including myocardial infarction, hypotension, pulmonary edema, and cardiac arrhythmia, have been described with such therapy. Concern over the toxicity of highdose interleukin-2 (IL-2) therapy has led to some clinicians excluding patients 70 years of age or over. We have treated 15 patients 70 years of age or over having an Eastern Conference Oncology Group (ECOG) performance status of 0 or 1, with therapy based on continuous infusion IL-2 18 MIU/sq m/24 hours for 72 hours. All patients underwent a pretreatment evaluation of cardiac status with a low-level stress or adenosine stress test. Cycles were typically repeated every 3 weeks for 4 cycles, then every 3-4 weeks thereafter. Patients were treated by oncology nurses in either the stem cell transplant (intermediate unit) or the oncology inpatient unit. Patient characteristics were: median age, 72 years (range, 70-83 years); tumor types: melanoma (10), kidney cancer (5); most common sites of disease: lung (11), lymph nodes (6), subcutaneous (3), liver (2); prior therapy included: none (8), outpatient IL-2 (5), other immunotherapy (4). Median number of cycles received: 3 (1-10). Most common toxicities were: fever, rigors, nausea, emesis, hypophosphatemia, and hypomagnesemia. Three patients required the use of dopamine for blood pressure support. Two patients declined further therapy. There were no treatment-related deaths. No patients required endotracheal intubation or transfer to an intensive care unit. One complete and 8 partial responses (60% response rate) have been seen. Responding sites include the lung, lymph node, intact kidney primary, and liver. Median survival has not been reached at over 14 months (range 3+-26+ months). Patients who are 70 years of age and older with an ECOG performance status of 0 or 1 are able to tolerate high-dose continuous infusion IL-2-based therapy and may respond to such treatment. PMID- 15778575 TI - Pharmacokinetics and clinical evaluation of 125I-radiolabeled humanized CC49 monoclonal antibody (HuCC49deltaC(H)2) in recurrent and metastatic colorectal cancer patients. AB - INTRODUCTION: CC49 is an antitumor monoclonal antibody that is promising for use in radioimmunoguided surgery (RIGS). However, the murine antibody has been limited by human antimouse antibody (HAMA) response and slow clearance. This study examined the pharmacokinetics and tissue localization of a humanized domain deleted CC49 antibody (HuCC49DeltaC(H)2 MAb) in humans. METHODS: Twenty-one patients with colorectal carcinoma were given 1 mg intravenous (I.V.) bolus of HuCC49DeltaC(H)2 MAb radiolabeled with 2 mCi (125)I after thyroid blockade. The level of circulating HuCC49DeltaC(H)2 MAb was measured daily as precordial counts using a handheld gamma-detecting probe. Each patient underwent an exploratory laparotomy on postinjection days 3-20. Gamma counts were measured at normal organs, aortic bifurcation (AB), and both clinically evident and occult tumors. RESULTS: Precordial and AB gamma counts showed an excellent linear correlation. HuCC49DeltaC(H)2 MAb followed a two-compartment pharmacokinetic model. Normal organs and AB showed similar exposures to HuCC49DeltaC(H)2 MAb, while HuCC49DeltaC(H)2 MAb favorably distributed into tumors from day 3. Intestinal and metastatic liver lesions showed the highest partition coefficients. All patients showed no HAMA response. DISCUSSION: C(H)2 region deletion of HuCC49DeltaC(H)2 MAb did not alter the pharmacokinetics compared to murine CC49. The favorable partition coefficient K of HuCC49DeltaC(H)2 MAb into tumors supports its use in RIGS. PMID- 15778576 TI - The role of 99mTc-tetrofosmin scintigraphy for staging patients with laryngeal cancer. AB - Diagnosis and staging of laryngeal cancer is currently based on physical examination, endoscopy, and imaging techniques such as computed tomography (CT) and/or magnetic resonance (MR) and histology. While imaging techniques have a pivotal role for defining the size of the primary tumor, they are less accurate for defining metastatic involvement of regional lymph nodes, especially if lymph nodes are smaller than 10-15 mm. The aim of this study was to comparatively assess the relevance of (99m)Tc-tetrofosmin scintigraphy for the staging of laryngeal tumors versus the CT scan. We evaluated the sensitivity of imaging with (99m)Tc-tetrofosmin in 28 consecutively enrolled patients with squamous cell laryngeal carcinoma. Total-body scintigraphy with 99mTc-tetrofosmin was performed preoperatively, and the results were compared to CT images of the neck and mediastinum. CT and (99m)Tc-tetrofosmin scintigraphy were equally sensitive (96%) in identifying the primary tumor. While CT was more sensitive for detecting metastatic lymph nodes (100% versus 50%), (99m)Tc-tetrofosmin scintigraphy was more specific (100% versus 56%; p < 0.04). The overall diagnostic capabilities of the two techniques for detecting lymph node metastases were comparable (Youden Index: J = 0.56 for CT and J = 0.50 for (99m)Tc-tetrofosmin scintigraphy). (99m)Tc-tetrofosmin scintigraphy is a useful complement to CT for staging laryngeal tumors, especially for detecting metastatic lymph nodes and distant metastases. PMID- 15778577 TI - High-dose continuous infusion plus pulse interleukin-2 and famotidine in metastatic kidney cancer. AB - High-dose continuous infusion interleukin-2 (IL-2) regimens generate a higher degree of lymphokine activated killer cell (LAK) cytotoxicity when tested against tumor cells in vitro and a higher rebound lymphocytosis in vivo than do bolus IL 2 regimens. Lymphocytes initially activated by continuous infusion IL-2 have increased cytotoxicity against cancer cells when they are subsequently pulsed with additional IL-2. Famotidine may enhance LAK cytolytic ability. Six patients with kidney cancer have been treated with a combination of famotidine 20 mg intravenous bid and continuous infusion IL-2 (18 MIU/sq m/24 hours) for 72 hours, followed by a 24-hour rest, then IL-2 18 MIU/sq m over 15-30 minutes. The most common metastatic sites were the lung, lymph node, and bone. Median number of cycles received = 5 (range, 3-8). The most common toxicities were fever, rigors, nausea/emesis, hypophosphatemia, hypotension, elevated creatinine, and metabolic acidosis. There were no treatment-related deaths, and no patients required intensive care admission. Two partial responses (33% response rate) have been seen. Median survival has not been reached at greater than 8 months. The combination of high-dose continuous infusion plus pulse IL-2 and famotidine is active in metastatic kidney cancer. An accrual of additional patients is needed to better assess the response rate. PMID- 15778578 TI - Proceedings of the 1st International Symposium on Radionuclide Therapy and Radiopharmaceutical Dosimetry. September 4-8, 2004, Helsinki, Finland. PMID- 15778580 TI - The radiobiology of conventional radiotherapy and its application to radionuclide therapy. AB - The linear-quadratic (LQ) model of radiobiological effect is well established in conventional, i.e., external beam, radiotherapy. Because the model is derived from sound biophysical principles, it is also emerging as the standard formalism for assessing biological responses for the whole range of radiotherapy treatments. A central feature of LQ methodology is the quantity known as the biologically effective dose (BED), which may be used to quantify the radiobiological impact of a treatment on both tumors and normal tissues. The BEDs commonly associated with conventional therapy may thus be compared to those expected from novel radiotherapy treatments, such as targeted radionuclide therapy. This approach also provides a mechanism for designing targeted treatments which are therapeutically equivalent to external beam treatments. In this paper the LQ methodology is outlined and worked examples are provided which demonstrate the tentative link between targeted radiotherapy doses and those used in conventional radiotherapy. The incorporation of an allowance for relative biological effectiveness (RBE) effects is also discussed. The complexity of the subject and the potentially large number of variables does place a restriction on overall predictive accuracy and the necessary caveats are outlined. PMID- 15778581 TI - A comparison of high- versus low-linear energy transfer somatostatin receptor targeted radionuclide therapy in vitro. AB - INTRODUCTION: The somatostatin analog [DOTA(0)-Tyr(3)]-octreotide (DOTATOC) has been widely used to target somatostatin receptor expressing tumors for therapy using radionuclides such as (90)Y or (177)Lu. AIM: This aim of this study was to compare the effects of DOTATOC labeled to high linear energy transfer (LET) alpha emitter (213)Bi and low-LET beta-emitter (177)Lu in vitro. MATERIALS AND METHODS: Somatostatin receptor (sstr)-positive cell line Capan-2 and sstr-negative control cell line A549 were used for the experiments. The effects of two exposure times using different radiation doses of high-LET alpha-emitter (213)Bi and low-LET beta-emitter (177)Lu were investigated using cell survival assay. The apoptotic effects were investigated using Cell Death Detection ELISA(PLUS)10x. The cumulated activity and the mean absorbed dose per unit cumulated activity were calculated using MIRD cellular Svalues. RESULTS: (213)Bi-DOTATOC had an approximately four times greater induction of apoptosis than (177)Lu-DOTATOC and a 100 times greater induction of apoptosis than nonradiolabeled DOTATOC. Nonspecific radiolabeled tetra-azacyclododecanetetra-acetic acid (DOTA) had a less pronounced effect on the cell survival and apoptosis, as compared to the sstr-specific radiolabeled DOTATOC. CONCLUSION: (213)Bi-DOTATOC is significantly more potent than (177)Lu-DOTATOC in vitro because of its high-LET alpha emission.(213)Bi-DOTATOC shows enhanced effects on mitotic and apoptotic cell deaths. PMID- 15778582 TI - The application of dose-rate volume histograms and survival fractions to multicellular dosimetry. AB - The distribution of therapeutic radiopharmaceuticals in volumes smaller than those that can be fully resolved by the imaging system, such as by PET and SPECT scanners, is usually assumed to be homogeneous. The aim of this study was to investigate the implications of such an assumption at a scale that can be defined as multicellular for heterogeneous activity localizations of (32)P, (90)Y, and (131)I. Dose-rate distributions from heterogeneous radioactivity uptakes have been calculated in cubic volumes of 1, 3, and 4 mm using the in-house software package DOVE. These have been studied by the use of dose-rate volume histograms, and the influence of the heterogeneous dose distribution on the treatment outcome has been analyzed by the calculation of Integral Survival Fractions. The results showed that the effect of the heterogeneous localization of the compound can be overridden by the amount of radioactivity administered. However, significant variations in the survival probability distributions have been observed, depending on the amount of initial activity considered, the activity configuration, the radionuclide, and the time over which the energy was deposited. It has been shown, for example, that the ability of longer-range beta emitters, such as (32)P and (90)Y, to invalidate heterogeneous dose-rate distributions may be negated by the decay rate of the radioactivity. PMID- 15778584 TI - A new method to obtain transmission images for planar whole-body activity quantification. AB - Dosimetry is important in the evaluation of new radiopharmaceuticals and in radionuclide therapy treatment planning. The base is an accurate in vivo measurement of the spatial and temporal distribution of the administered radioactivity. This can be made by planar whole-body activity quantification using the conjugate-view method where a geometrical-mean image is corrected for attenuation by a measured transmission image. We present a new method to measure whole-body transmission images using the X-ray unit on a single photon emission computed tomography/computed tomography (SPECT/CT) scintillation system. METHOD: Two general problems are associated with this method: (1) the divergence of the X ray beam and (2) the scaling of the resulting image. We have applied linear transformations and scale by a system-generated factor to obtain images corresponding to a radiological depth at 70 keV. We validated the method by measurements of the radiological depth for different types of phantoms, and in terms of activity quantification accuracy when applying the method for attenuation correction. RESULTS: An accuracy of within +/-4% was found for both the radiological depth measurements and for the activity-quantification measurements. CONCLUSIONS: The advantage with the proposed method is that it is fast (2 minutes for a 2-minute scan), it has good spatial resolution and signal to-noise ratio, and it is easy to carry out for the patient. PMID- 15778583 TI - Whole-body dosimetry for targeted radionuclide therapy using spectral analysis. AB - The whole-body dose (WBD) is routinely calculated for targeted radionuclide therapy (TRT). The aim of this work was to investigate the feasibility of using spectral analysis (SA) for the automatic delineation of decay phases, and consequently, the calculation of the WBD given a whole-body (WB) time-activity curve (TAC). SA characterizes the TAC as an arbitrary sum of exponential functions determined by fitting the data with a non-negative least-squares (NNLS) algorithm. The cumulated activity (CA) is calculated analytically as the integral of the fitted curve while the number of phases describing the kinetics of the radiopharmaceutical and the half-lives of the phases can be determined from the spectrum. The uncertainty associated with the estimation of the WBD can be obtained using bootstrap techniques. SA was applied to WB TACs from (186)Re-HEDP and (131)I-mIBG therapies. The results were compared to results obtained using a semiempirical method and showed good agreement in the calculated WBDs. Bootstrapping with resampling on a subset of patients from the two therapies showed much larger coefficient-ofvariation (CV) for the (186)Re-HEDP TACs than for the (131)I-mIBG therapies. We concluded that SA provides a fast, accurate, and reproducible method to obtain WBDs and accurate estimates of the parameters describing the radiotracer kinetics. The method could be extended to other dosimetric applications. PMID- 15778585 TI - Monte Carlo modeling of gamma cameras for I-131 imaging in targeted radiotherapy. AB - INTRODUCTION: Dosimetric studies for targeted radiotherapy require the quantification of activity from scintigraphic images. Quantitative imaging is difficult to achieve because of several effects that can lead to errors in activity estimates, some of which are more apparent when I-131 is considered as a source. An evaluation of these phenomena was performed by modeling the gamma camera and its behavior using Monte Carlo simulations. Two gamma cameras were modeled: DST-XLi and Millennium VG Hawk-Eye (GEMS), and two Monte Carlo codes were used: MCNP (LANL) and GATE (openGate collaboration). GATE is a dedicated single photon emission computed tomography/positron emission tomography (SPECT)/(PET) software based on Geant4 (CERN, Geneve). MATERIALS AND METHODS: Gamma-camera modeling was performed in 2 steps: first without a collimator, then with a high-energy, all-purpose (HEAP) collimator according to the specifications given by the manufacturer (the simulation took the hexagonal shape of collimator holes into account). Simulated and measured energy spectra from point sources in air were compared (with or without a collimator). Spatial resolution was obtained from line sources in air at various distances from the detector heads. The photons detected in the 20% energy window from a point source were analyzed in order to determine the amount of primary photons, scattered photons (in the collimator), and septal photons (i.e., photons that crossed the collimator septa without interacting). RESULTS: Both codes agree well with experimental measurements for the two gamma cameras considered in this study. This allowed us to validate gamma-camera modeling and also served as a benchmark of GATE (new code) versus MCNP (reference code). As shown previously by Dewaraja et al., septal penetration is an important source of image degradation when HEAP collimators are used for I-131 imaging. With the DST-XLi, and for a point source in air, our simulations have shown that 53% of scattered (30%) and septal penetration (23%) photons are detected in the 20% window. CONCLUSION: The modeling of two gamma cameras (DST-XLi and Millennium VG Hawk-Eye) has been performed with two Monte Carlo codes (MCNP and Gate). Results obtained with the two Monte Carlo codes agree well with experimental results. As already indicated by several authors, septal penetration and scattered photons in the collimator have a major impact on I-131 scintigraphic imaging. PMID- 15778587 TI - The LundADose method for planar image activity quantification and absorbed-dose assessment in radionuclide therapy. AB - A new method for absorbed-dose assessment in radionuclide therapy is presented in this paper. The method is based on activity quantification by the conjugate-view methodology, applied to serial whole-body, anterior-posterior, scintillation camera scans. The quantification method is an extension of previous studies, and includes separate corrections for attenuation, scatter, and overlapping organs. Further development has now been undertaken to take into account the capabilities of new dual-head camera systems with a built-in X-ray tube for anatomical imaging. Furthermore, the modeling of time-activity data is included, and dosimetric calculations based on the formalism by the Medical Internal Radiation Dose (MIRD) committee. To streamline absorbed-dose assessments for a large number of patient studies, the programs for quantification, image registration, and absorbed-dose calculations have been embedded in an envelop program termed LundADose, where calculations, to a great extent, are performed automatically. Evaluation of the whole-body activity quantification is performed for patients undergoing radioimmunotherapy by monoclonal antibodies labeled with (111)In or (90)Y. PMID- 15778586 TI - Dosimetry calculations on a tissue level by using the MCNP4c2 Monte Carlo code. AB - OBJECTIVE: The aim of this study was to develop a MCNP4c2-code and to further refine the small-scale anatomy intestinal dosimetry model based on a EGS4-code developed by Jonsson et al.(1,2) METHOD: The small intestine was modeled as a hexagonal tube system and includes cross-dose contribution from activity in nearby intestine loops. The model includes villi (height, 500 microm), radiosensitive crypt cells (height, 150 microm), and an overlying mucus layer of thicknesses (5-200 microm). The developed intestinal model used in either of the two Monte Carlo codes make it possible to calculate S-values and subsequent mean absorbed dose to the radiation-sensitive crypt cells in the small intestinal wall by considering contributions from the self-dose and from the cross-dose from nearby intestinal loops. Results are given for monoenergetic electrons and photons and for full decay schemes of (99m)Tc, (111)In, (131)I, (67)Ga, (90)Y, and (211)At. RESULTS: Results show that the cross-dose from nearby intestinal loops is significant, and that the fraction of cumulated activity in the intestinal wall contents is important for accurate absorbed-dose estimation. CONCLUSION: It is evident from our study that previous Medical Internal Radiation Dose (MIRD) and International Conference on Radiological Protection (ICRP) models tend to overestimate the absorbed dose to the wall. Our work on the gastrointestinal tract model includes several noticeable refinements, as compared to the MIRD- and ICRP model, and the "onion shell" geometry can easily be transferred to similar geometrical dosimetry applications. PMID- 15778588 TI - Determination of individual S-values for 131I using segmented CT data and the EGS4 Monte Carlo code. AB - In individual voxel phantoms, which were segmented from whole-body computed tomography (CT) scans, S-values were calculated for (131)I using the EGS4 Monte Carlo code and compared to Medical Internal Radiation Dose (MIRD) S-values, which were derived from transport calculations in idealized mathematical phantoms. The individually calculated S-values agree very well with the MIRD values for organs, which are source and target simultaneously, when individual organ-mass corrections are applied to the MIRD values. For different source-target combinations, large deviations up to 184% were found. The contribution of the gamma-absorbed fractions to the total dose, however, is small ( approximately 4%). We conclude, therefore, that individual transport calculations in radionuclide-targeted therapies are not necessary for macroscopic dose estimates. Reliable dosimetry is reduced to the problem of accurate activity determination in vivo. PMID- 15778589 TI - Estimation of patient dose from radiopharmaceuticals using voxel models. AB - The aim of this study was to demonstrate the advantages of patient dosimetry using voxel models and to present sets of dose estimates for patients of different gender and size. These models offer greater realism with respect to organ shape and topology than the well-established Medical Internal Radiation Dose (MIRD)-type mathematical models. At the National Research Centre for Environment and Health (GSF), specific absorbed fractions have been previously calculated for 4 male and 3 female voxel models, representing different age and stature, for a wide range of source organs. For this study, estimates both for established and new radiopharmaceuticals were performed using biokinetic data from International Commission on Radiological Protection (ICRP). The above calculations allowed for comparison to the MIRD technique in relation to the resulting absorbed organ and effective doses. Furthermore, data sets representing a range of voxel phantoms were investigated. It was found that dose differences among the voxel models can amount up to a factor of 3. PMID- 15778590 TI - Testicular uptake and radiation dose in patients receiving Zevalin and Pretarget CC49Fusion protein. AB - OBJECTIVE: Radiation dose to the testes from radionuclide therapies is of concern. This study evaluated image-quantification methods for testicular uptake in a phantom and in patients. METHODS: A 50-mL vial and a large water tank were used to simulate testes and the body, respectively. Activity concentration in the vial and water tank was prepared to generate testes-to-background concentrations of 1.3 and 1.1. Five male lymphoma patients who received a Zevalin (Biogen Idec, Cambridge, MA) regimen and 6 male colorectal cancer patients who received a Pretarget (Neo Rx, Seattle, WA) CC49Fusion protein were evaluated. Testicular activity was quantified using two methods: (1) geometric-mean, background corrected testicular region of interest (ROI) counts as a fraction of body counts without explicit attenuation correction (Zevalin Kit); (2) background-corrected anterior testicular ROI counts with attenuation correction using known depth in the phantom and CT depth in patients. RESULTS: In the phantom study, Method 1 underestimated 49% and 39%, at image contrast of 1.3 and 1.1, respectively. Quantification was improved using Method 2 (7% for a 1.3 contrast, -17% for a 1.1 contrast). Method 2 was used in patients because background-corrected posterior ROI counts were statistically unreliable due to poor image contrast. In patients receiving Zevalin, the median peak percent injected dose (%ID)/testis was 0.10 (range, 0.08-0.18) with a median biologic half-time (T(bio1/2)) of 156 (range, 91 4200) hours. The median dose was 2.4 (range, 1.5-3.6) Gy/GBq, compared to the originally reported mean dose of 9.1 (range, 5.4-11.4) Gy/GBq (Zevalin package insert). In patients receiving the Pretarget CC49Fusion protein, the median peak %ID/testis was 0.22 (range, 0.05-0.29) with a median T(bio1/2) of 44 (range, 37 64) hours. The median dose was 0.84 (range, 0.3-1.2) Gy/GBq. CONCLUSION: This study found that testicular doses from Zevalin were much lower than that originally reported in the package insert. The median testicular dose from Pretarget CC49Fusion protein was less than half that of the median testicular dose from Zevalin. PMID- 15778593 TI - Nontuberculous (atypical) mycobacterial keratitis after LASIK: current status and clinical implications. AB - PURPOSE: Corneal infection with unfamiliar organisms such as nontuberculous (atypical) mycobacteria after laser in situ keratomileusis (LASIK) can be a significant clinical problem, and mismanagement of such corneal infection in an otherwise healthy, young individual can lead to significant medicolegal issues for the refractive surgeon. Because nontuberculous (atypical) mycobacterial keratitis is the most common infection after LASIK, the refractive surgeon should be aware of all aspects of this dreaded infection to be better prepared to manage such cases. METHODS: Literature search. RESULTS/CONCLUSIONS: This article provides a comprehensive compilation of all reported cases of nontuberculous (atypical) mycobacterial keratitis in the English literature and provides some useful recommendations for the clinical management of such corneal infection after LASIK. PMID- 15778594 TI - Forty years of changing indications in penetrating keratoplasty in Israel. AB - PURPOSE: To examine the leading indications for keratoplasty and identify the changing trends in the past 40 years in Israel. METHODS: Pathology reports of all penetrating keratoplasties (PKPs) performed at Hadassah-Hebrew University Hospital from 1961 to 2000 were reviewed. We evaluated the indications for keratoplasty in each decade between the years 1961 and 2000. RESULTS: During the 40-year period, a total of 1681 PKPs were performed. Keratoconus (n = 478, 28.4%) was the most common indication, followed by graft failure (n = 226, 13.4%), pseudophakic corneal edema (PCE) (n = 142, 8.4%), herpetic infections (n = 125, 7.4%), nonherpetic infections (n = 114, 6.8%), scarring (n = 113, 6.7%), and trauma (n = 110, 6.5%). The 7 groups account for approximately 77.5% of all keratoplasties performed. Ocular infections were the most common indications before 1970 and have been declining ever since. Keratoconus became the leading indication in the past 30 years. In the past decade, of 663 keratoplasties, keratoconus was the most common indication (56, 38.6%) followed by graft failures/rejections and PCE (the second most common indication between the years 1981 and 1990). CONCLUSIONS: Keratoconus was the leading indication for keratoplasty in our series. There was a decreasing trend in PK for ocular herpetic infections during the decades. The rate of PCE decreased while failed graft became the second most common indication for PKP during the past decade. PMID- 15778595 TI - White-to-white corneal diameter: normal values in healthy humans obtained with the Orbscan II topography system. AB - PURPOSE: The corneal horizontal diameter (white-to-white) is abnormal in diseases like microcornea, relative anterior microphthalmos, and corneal dystrophies. Because normal values are described imprecisely in the literature, the purpose of this study was to reevaluate the horizontal corneal diameter as a scientific parameter. METHODS: The horizontal corneal diameter was measured with the Orbscan II system in 370 right eyes and 373 left eyes of 390 healthy white subjects aged 10-80 years. There were 148 female subjects and 242 male subjects. Each measurement was repeated twice. Differences in gender, between right and left eyes, and age-related alterations were analyzed statistically. RESULTS: The average corneal diameter was 11.71 +/- 0.42 mm. The average corneal diameter was 11.77 +/- 0.37 mm in males compared with 11.64 +/- 0.47 mm in females. The resulting normal ranges were 11.04 to 12.50 for males and 10.70 to 12.58 mm for females. Differences in gender were not significant in the t test for independent samples (P = 0.071). There were no statistically significant differences between right and left eyes in the t test for dependent samples (P = 0.16). Corneal diameters decreased slightly with age. CONCLUSIONS: With the obtained normal values, more precise determination of microcornea and macrocornea will be possible in the future. The horizontal corneal diameter was not significantly greater in males than in females. Further studies are needed to show the reasons for the age-related decrease in measurements. PMID- 15778596 TI - Phototherapeutic keratectomy in treating keratomycosis. AB - PURPOSE: To evaluate the efficacy of using phototherapeutic keratectomy (PTK) in the treatment of superficial keratomycosis. METHODS: We studied 9 patients with superficial keratomycosis that had infiltrated less than half of the corneal thickness and responded poorly to topical antifungal therapy. Using a 193-nm excimer laser, we performed PTK to try to eradicate the infiltrates and facilitate antifungal therapy. We compared our study group findings with those of a control group made up of 31 cases of keratomycosis, also involving infiltration of less than half of the corneal thickness that had been treated with traditional surgical procedures and topical antifungal agents. RESULTS: Using PTK and short term antifungal eyedrops, we were able to eradicate keratomycosis without recurrence in all 9 study patients. Much less time was needed to treat the PTK group (12.9 +/- 3.6 days) than the control group (40.8 +/- 26.4 days) (P <0.05). After PTK, the ablated area underwent rapid reepithelialization (average, 3.6 +/- 1.8 days). Final vision ranged from 20/200 to 20/20. The PTK group had an average visual improvement of 2.9 +/- 2.1 lines, which was significantly better than final improvement found in the control group (average, 0.6 +/- 1.7 lines) (P < 0.05). PTK complications included mild corneal haze, astigmatism, and thinning cornea. CONCLUSIONS: Because PTK can shorten treatment time, hasten reepithelialization, and restore reasonably good vision, it can be a valuable therapeutic alternative for superficial keratomycosis, especially in instances in which there is poor response to treatment by topical antifungal agents alone. PMID- 15778597 TI - Scleral contact lenses: the expanding role. AB - PURPOSE: The purpose of this study was to describe the current indications for scleral contact lens (ScCL) management at Moorfields Eye Hospital, London. METHODS: A database of 1003 patients (1560 eyes) seen between September 1999 and May 2003, either assessed for an ScCL trial or to follow up previously issued lenses, was analyzed. RESULTS: The major contact lens indication groups were keratoconus or other primary corneal ectasia (PCE), corneal transplant, and ocular surface disease (OSD). The 3 main small groups were aphakia, myopia, and ptosis. A total of 538 patients continued lens wear in 1 or both eyes, for a total of 808 eyes. The total numbers of eyes for each contact lens indication were PCE, 496 (61.4%); corneal transplant, 150 (18.6%); OSD, 91 (11.4%); aphakia, 17 (2.1%); myopia 21 (2.6%); ptosis 14 (1.7%); and a miscellaneous indication, 19 (2.2%). The percentages were similar to those of the total group of 1560 eyes. Seven hundred twenty lenses (89.1%) were rigid gas permeable (RGP), 708 were nonventilated, and 12 were fenestrated; 88 (10.9%) were ventilated polymethylmethacrylate or ptosis props. CONCLUSIONS: ScCLs continue to play a significant role in the management of corneal abnormalities, especially PCE, corneal transplant, and OSD. The great majority of lenses in use were nonventilated RGP designs. PMID- 15778598 TI - Corneal endothelium evaluation after phacoemulsification with continuous anterior chamber infusion. AB - PURPOSE: To determine if the use of continuous anterior chamber infusion (CACI) during conventional phacoemulsification can damage the corneal structure by adding an additional hydrodynamic stress to the corneal endothelium intraoperatively and to use this option routinely in small-incision phacoemulsification to soften its learning curve. METHODS: We performed a prospective, interventional, case series of patients with cataract who underwent phacoemulsification. Patients were previously classified into 2 groups based on whether CACI was used (group 2) intraoperatively or not (group 1). Pachymetry and endothelial cell (EC) density were evaluated before and after surgery, and data were compared between the 2 groups. Eyes with values of EC density less than 1000 cells/mm or showing previous corneal structural alterations were excluded from the study. RESULTS: Sixty-seven eyes were included in our study from September 2002 to January 2003. Group 1 included 31 eyes (mean pachymetry 520.4 microm and EC density 2883), and group 2 included 36 eyes (mean pachymetry 519.0 and EC density 2627). Multivariate analysis of postoperative data following a multiple regression model showed an increase in pachymetry for both groups at 24 hours after surgery that progressively diminished until the last visit 4 weeks postoperatively, and no statistically significant differences were found between the groups. EC density displayed an abrupt loss for both groups, especially for group 2, that reached statistical significance at day 7 after surgery (P = 0.020). EC density experienced an increase for both groups in follow-up visits, and it was not statistically different at last postoperative control (P = 0.361). CONCLUSIONS: : The trend in modern cataract surgery is to reduce postoperative ocular trauma, and this can be achieved by small-incision phacoemulsification. But it requires a learning curve that we believe can be softened by using some surgical maneuvers such as continuous anterior chamber infusion, which minimizes the risk for anterior chamber collapse during the intervention. Comparing postoperative pachymetry and EC density between group 1 and 2, we found that the use of CACI does not add any additional damage to corneal structures because the initial statistically significant EC loss at day 7 postoperatively has a prompt recovery in subsequent visits, and then the EC loss remains similar in both groups. Transition from conventional to bimanual phacoemulsification can be simplified by using CACI, and it does not increase postoperative corneal damage. PMID- 15778599 TI - Delayed healing of corneal epithelium after phototherapeutic keratectomy for lattice dystrophy. AB - PURPOSE: To evaluate the time period necessary for complete epithelial healing after phototherapeutic keratectomy (o-PTK) carried out for various superficial corneal opacities. SUBJECTS AND METHOD: A total of 197 eyes were divided into 9 groups: group 1, Cogan dystrophy including recurrences (n = 15); group 2, Reis Bucklers dystrophy including recurrences (n = 12); group 3, granular dystrophy including recurrences (n = 63); group 4, lattice dystrophy including recurrences (n = 19); group 5, macular dystrophy including recurrences (n = 10); group 6, herpetic scars (n = 5); group 7, corneal scars of nonherpetic origin (including scrofulous, traumatic, central keratoconus, post-pterygium surgery) (n = 31); group 8, Salzmann nodular degeneration (n = 22); and group 9, miscellaneous (such as bullous keratopathy, acute chemical burn, corneal degeneration) (n = 20). After o-PTK, patients were examined daily at the slit lamp using fluorescein and blue light. The time period necessary for complete healing of the epithelial defect was compared among these groups. Delayed healing was considered where the epithelium was not closed after 7 days. RESULT: One hundred sixty-one eyes (95%) healed within 7 days. Overall, 63%, 80%, and 85% of epithelial defects were closed within 3, 4, and 5 days, respectively. Out of 9 eyes that had delayed healing, 6 eyes (67%) belonged to lattice dystrophy category. Mean time taken for healing in group 4 (8.6 +/- 8.4 days) was significantly longer than those in group 1 (3.0 +/- 1.5 days, P = 0.009), group 2 (3.7 +/- 3.1 days, P = 0.03), group 3 (3.1 +/- 1.5 days, P = 0.001), group 5 (2.7 +/- 0.8 days, P = 0.01), group 7 (3.6 +/- 2.4 days, P = 0.007), group 8 (3.3 +/- 1.3 days, P = 0.009), and group 9 (3.0 +/- 1.9 days, P = 0.011). CONCLUSION: Eyes with lattice corneal dystrophy suffered from delayed epithelial healing after o-PTK. In addition to adequate counseling, these patients should be followed up closely until complete closure of the epithelium to avoid ulceration, scarring, or even infection. These eyes might need additional treatment such as hyaluronic acid drops, autologous serum drops, simultaneous amniotic membrane patching, or even temporary lateral tarsorrhaphy. PMID- 15778600 TI - The incidence of fungal keratitis and endophthalmitis following penetrating keratoplasty. AB - PURPOSE: To determine the incidence of postkeratoplasty fungal endophthalmitis and keratitis at the New York Eye and Ear Infirmary. To determine whether there is a relationship between culture-positive corneoscleral donor material and postoperative infection. METHODS: The microbiologic records of corneoscleral donor rims submitted for culture following penetrating keratoplasty at the New York Eye and Ear Infirmary between January 1998 and January 2003 were reviewed. The incidence of rim cultures positive for fungi was tabulated. Clinical outcome measures were recorded for each patient receiving corneal donor tissue. RESULTS: Of 2466 donor corneoscleral rims cultured during the study period, 344 were positive for microbial growth (13%). Of those rims with positive cultures, 28 (8.6%) were positive for fungus. All fungi cultured were Candida species. Four of the 28 recipient eyes (14%) who received contaminated donor material went on to develop postkeratoplasty fungal infections. There were no cases of fungal infection in any postkeratoplasty patients in the absence of contaminated donor rims during the study period. Overall, there was a 0.16% incidence of fungal infection (4/2466) following penetrating keratoplasty. There were 18 positive donor rims identified in the first 4 years of the study, but there were 10 cases in the last 10 months of the study. CONCLUSIONS: The overall incidence of fungal infection following penetrating keratoplasty is low, but all cases in our study were associated with positive rim cultures. Whether prophylactic antifungal therapy would be of any benefit in the presence of a positive corneoscleral rim culture has not yet been determined. PMID- 15778601 TI - Opinions on risk factors and management of corneal graft rejection in the United kingdom. AB - PURPOSE: To determine the opinions regarding risk factors and practice preferences for corneal graft rejection by members of the Bowman Club (UK) and to compare them with those of members of the Castroviejo Society (USA). METHODS: A questionnaire was sent in 1999 to members of the Bowman Club (UK), who were responsible for two thirds of all corneal grafts undertaken annually. The survey included 8 questions identical to those given to members of the Castroviejo Society (USA) in a survey carried out in 1989. RESULTS: Thirty-six out of 40 surgeons replied. Factors considered by respondents to be high risk for corneal graft rejection were previous corneal graft rejection in the operated eye (97%), significant corneal vessels (97%), and previous herpetic eye disease (94%). The preferred routine preoperative treatment in "high-risk" patients included no treatment (47%), topical corticosteroids (33%), and oral prednisolone (22%). In postoperative "high-risk" patients, 100% of surgeons used topical and 42% used oral corticosteroids. Immune suppression agents were used by 44% of respondents, the majority (92%) using cyclosporine A. In previous herpes simplex patients, 47% of surgeons used oral and 14% used topical antivirals preoperatively, whereas 75% used oral and 47% used topical postoperatively. CONCLUSION: This study documents the perceived risk factors and management of corneal graft rejection by corneal surgeons in the UK. It showed wide variation in practice preferences, allowing individual surgeons a comparison with peer practice. It highlights the need for greater use of postoperative antiviral prophylaxis in the presence of previous herpetic corneal pathology. PMID- 15778602 TI - Ocular surface squamous neoplasia: a standard of care survey. AB - PURPOSE: To determine the current standard of care in the treatment of ocular surface squamous neoplasia (OSSN) with particular regard to topical chemotherapeutic agents including mitomycin C, 5-fluorouracil, and interferon alpha2b. METHODS: A mail survey of the 325 physician members of the Ocular Microbiology and Immunology Group. RESULTS: The survey response rate was 38%, and the average number or cases of OSSN managed by respondents was 6.5 per year. A majority (54%) believed sufficient evidence existed to justify the use of mitomycin C in the treatment of OSSN, and fewer felt the published literature justified the use of 5-fluorouracil or interferon (11% and 21%, respectively). About one-half of responding physicians always perform a biopsy before institution of topical therapy. The reported use of topical chemotherapy as an adjunct to surgical excision increased with the size of the lesion; 45% of the respondents utilize topical therapy along with surgery for lesions greater than 8 mm in diameter. CONCLUSION: The standard of care for the treatment of ocular surface squamous neoplasia appears to have shifted from surgery toward the use of topical chemotherapeutic agents as adjuncts to surgery or even as sole therapy, despite a paucity of long-term studies in the published literature. Longer-term observational studies and prospective masked and controlled studies are indicated to determine the optimal treatment(s) for squamous neoplasia of the ocular surface. PMID- 15778603 TI - Personality in keratoconus in a sample of patients derived from the internet. AB - PURPOSE: Keratoconus is a corneal disease that manifests itself in young adulthood and decreases vision. Anecdotally, keratoconus patients are commonly described as having unusual personality characteristics; however, previous researchers have not been able to define a unique keratoconus personality profile. We sought to better define the personality of patients with keratoconus. METHODS: The Millon Behavioral Health Inventory (MBHI) is a psychological instrument designed for use with physically ill patients to assess factors relevant to their medical care. The MBHI may aid in the evaluation of keratoconus patients and provide information relevant to the doctor-patient relationship. Patients with keratoconus were recruited through the World Wide Web. Surveys were conducted by mail, with all responses returned anonymously. One hundred fifty three keratoconus patients were compared with the MBHI age-matched norms, generated from medical and nonmedical samples of adults. Subgroups of keratoconus patients were compared based on their self-reported rating of the daily burden of keratoconus. RESULTS: Keratoconus patients scored significantly lower (chi, P <0.05) compared with the normative population on the respectful coping style scale, and their scores were not significantly associated with a history of penetrating keratoplasty or their rating of the effect of keratoconus on their lives. This suggests that keratoconus patients are less respectful of practitioners, uncooperative, and noncompliant with treatment plans. Practitioners may look unfavorably on patients who exhibit these characteristics. This would also explain why the clinical perception of keratoconus patients persists, but other personality inventories have not defined a distinct keratoconus profile. CONCLUSIONS: These data from the MBHI, showing that keratoconus patients score lower on the respectful coping style scale, may explain eye care practitioners' perceptions about these patients' personality characteristics. PMID- 15778604 TI - Surgical management of scleral defects. AB - PURPOSE: To evaluate the results of surgical management of scleral defects using fascia lata, cornea, and sclera as graft materials. METHODS: The scleral defects of 8 patients were repaired surgically. Six had previous intraocular surgery, 1 had eye trauma, and the other had both intraocular surgery and trauma. Corneoscleral grafts were used in 3 eyes, scleral grafts in 4, and fascia lata was used in 1 eye. RESULTS: In 7 patients, we achieved the desired cosmetic and tectonic success. During the follow-up period, a recurrent defect was seen in 1 patient, and a second operation was required. CONCLUSIONS: Surgical techniques using fascia lata, corneal, and scleral grafts are effective for scleral defects. PMID- 15778605 TI - MHC class I and II antigens as targets of rejection in penetrating keratoplasty in low- and high-risk mouse eyes. AB - PURPOSE: To determine the extent to which expression of major histocompatibility complex (MHC) class I and II molecules contributes to rejection of orthotopic corneal transplants in mice. METHODS: Full-thickness corneas, prepared from eyes of normal C57BL/6 (B6) and BALB/c mice and from B6 mice in which the class II gene I-A or the beta2-microglobulin (beta2mu) gene was disrupted, were placed orthotopically in low- or high-risk eyes of BALB/c (fully incompatible), BALB.B [minor histoincompatible (H) only], and bm12 (class II only disparate) recipients. BALB/c grafts were placed in low-risk eyes of normal B6 and B6 mice with disrupted H-2 DMalpha genes. Graft survival was judged by clinical examination. RESULTS: Recipient-identical class II, but not class I, molecules on fully allogeneic corneas grafted to low-risk beds promoted graft rejection. Allogeneic class II molecules on fully allogeneic corneas placed in high-risk beds promoted graft rejection more strongly than did allogeneic class I molecules. Neither allogeneic class I molecules nor recipient-identical class II molecules on grafts placed in high-risk beds contributed to graft outcome. Mice deficient in H-2 DMalpha failed to reject fully incompatible cornea grafts. CONCLUSIONS: On corneal allografts, where minor H antigens are the major barriers to acceptance, allogeneic class II molecules promote rejection if the graft is placed in high-risk eyes, whereas recipient-type class II molecules promote rejection if the graft is placed in low-risk eyes. Allogeneic class I molecules make a minor contribution to rejection only if the grafts are placed in high-risk eyes. PMID- 15778607 TI - Femtosecond laser posterior lamellar keratoplasty: a laboratory model. AB - PURPOSE: To evaluate feasibility of femtosecond laser application in posterior lamellar keratoplasty. METHODS: To evaluate the laser's effectiveness through opaque corneas, anterior corneal caps were resected from opaque corneas induced with 80% acetone solution. To evaluate the femtosecond laser posterior lamellar keratoplasty surgical procedure, human corneoscleral rims were mounted on an artificial anterior chamber. After corneal pachymetry, the femtosecond laser was used to create a 6-mm-diameter, 200-microm-thick endostromal lenticule. Access to the lenticule was provided by a small perilimbal surface opening, also created by the laser. The lenticule was removed using a pair of corneal forceps. A donor lenticule of similar dimensions was created, its endothelial surface coated with viscoelastic, inserted, and positioned on the recipient bed. Two sutures were placed to seal the small surface opening. RESULTS: The femtosecond laser produced an effective and smooth dissection through opaque corneas even at deeper settings. Graft transplantation was fairly simple and effective. CONCLUSION: Femtosecond laser posterior lamellar keratoplasty is a procedure that may provide an alternative to penetrating keratoplasty or the technically challenging manual posterior lamellar keratoplasty. PMID- 15778606 TI - A morphologic study of Fuchs dystrophy and bullous keratopathy. AB - PURPOSE: To describe the morphologic features of Fuchs corneal dystrophy and compare them with those of bullous keratopathy. METHODS: This was an observational case series. The study group consisted of 32 corneal buttons with a diagnosis of Fuchs dystrophy and the comparison group consisted of 22 corneal buttons with bullous keratopathy. Morphologic analysis was performed of corneal buttons from patients with the clinical diagnosis of Fuchs dystrophy or bullous keratopathy by light and electron microscopy. RESULTS: The main outcome measure was identification of degenerated keratocytes, granular material in and around keratocytes, and lipid keratopathy. The overall morphologic features of Fuchs dystrophy and bullous keratopathy are similar to those described in previous literature. A high proportion of keratocytes exhibited degenerative changes (78.9% in Fuchs dystrophy versus 50.5% in bullous keratopathy). Granular material was identified in and around variably degenerated keratocytes in all cases of Fuchs dystrophy and in 14 of 22 (64%) of the corneas with bullous keratopathy. The percentage of keratocytes with granular deposits was higher in Fuchs dystrophy corneas as compared with corneas with bullous keratopathy (51.7% versus 14.1%, P < 0.0005). Lipid keratopathy was a common occurrence in both Fuchs dystrophy and bullous keratopathy (23/32 [72%] versus 12/22 [55%]). CONCLUSIONS: Histopathologic changes in the corneal stroma and keratocytes occur in Fuchs dystrophy. Secondary lipid keratopathy ensues and may contribute to corneal haze. A higher proportion of keratocytes in Fuchs dystrophy have granular deposit than in bullous keratopathy. That a high proportion of keratocytes had degenerative changes in both Fuchs dystrophy and bullous keratopathy suggests that keratocytes may degenerate secondary to altered stromal microenvironment because of endothelial cell loss. PMID- 15778608 TI - Selective uptake of iron oxide by rabbit conjunctival lymphoid follicles. AB - PURPOSE: To determine whether conjunctival lymphoid follicles are preferential sites for uptake of foreign material in the preocular tear film. METHODS: Iron oxide suspension was applied to the eyes of New Zealand white rabbits for selected times, and the conjunctiva was examined histochemically for the presence of iron. RESULTS: Iron was observed by histochemical staining within conjunctival follicle-associated epithelium at 1 hour and deep within the follicles at 4 hours after exposure. Iron was not seen within nonfollicular conjunctival epithelium or underlying substantia propria at any time after iron oxide application. CONCLUSIONS: Iron oxide in the preocular tear film is taken up preferentially by conjunctival lymphoid tissue, supporting the hypothesis that mammalian conjunctival lymphoid follicles may participate in the acquired immune response to pathogens in the preocular tear film. PMID- 15778609 TI - Human leukocyte class I antigen and beta2-microglobulin expression in conjunctival dysplasia, carcinoma in situ, and squamous cell carcinoma. AB - PURPOSE: Malignant transformation of cells is frequently associated with abnormalities in HLA expression. These abnormalities may play a significant role in the clinical course of the disease because the cellular immune response to tumors relies on concomitant recognition of tumor antigens with self-HLA molecules. Since HLA molecules mediate interactions of tumor cells with specific receptors on T and natural killer (NK) cells, tight control of expression of HLA molecules is critical for initiation and implementation of an effective cellular immune response. There is not much information on HLA class I antigens in conjunctival carcinomas. We studied the immunoexpression of HLA class I antigen and beta2-microglobulin (beta2-m) in conjunctival dysplasias and conjunctival squamous cell carcinomas. METHODS: HLA class I antigen and beta2-m expression were analyzed in 10 conjunctival dysplasias, 6 carcinomas in situ, and in 11 conjunctival carcinomas by immunoperoxidase staining with monoclonal antibodies to HLA class I antigen and beta2-m on the archival clinical samples. Immunoanalysis was done according to the International Histocompatibility Working Group Project Description. RESULTS: HLA class I antigen and beta2-m were heterogeneous in 10 conjunctival dysplasias, negative in 6 carcinomas in situ, and 11 conjunctival squamous cell carcinomas. The difference of immunoexpression for HLA class I and beta2-m among the 3 groups was statistically significant (P <0.001). CONCLUSIONS: HLA class I antigens and beta2-m are decreased in conjunctival dysplasias and negative in carcinomas in situ, and carcinomas. The findings support the role of cytotoxic T lymphocyte-mediated control of tumor growth in the clinical course of conjunctival squamous cell carcinomas. PMID- 15778610 TI - Transient band-like keratopathy after treatment for seborrheic dermatitis. AB - PURPOSE: To describe a case of transient band-like keratopathy after ocular exposure to fluocinonide cream and ketoconazole shampoo. DESIGN: Observational case report. RESULTS: A 40-year-old patient presented with acute pain, photophobia, lacrimation, and redness in 1 eye. The symptoms began while using fluocinonide cream and ketoconazole shampoo as treatment of seborrheic dermatitis of the scalp. Examination revealed white clumpy deposits in a horizontal band across the inferocentral corneal epithelium and conjunctival hyperemia. Corneal scrapings revealed no cells or organisms, and culture was negative. The lipophilic deposits dissolved during slide fixation and processing. With conservative treatment the deposits resolved in 3 days. CONCLUSION: We present a case of transient band-like corneal deposit, a novel complication of fluocinonide and ketoconazole exposure. PMID- 15778611 TI - Mycobacterium keratitis after laser in situ keratomileusis. AB - PURPOSE: We present a case of Mycobacterium keratitis after laser in situ keratomileusis (LASIK). METHOD: A retrospective case report. RESULTS: A 40-year old woman developed Mycobacterium chelonae keratitis in the right eye 2 weeks after bilateral LASIK. The diagnosis was made by acid-fast staining of corneal scrapings on admission and confirmed by culture results 2 weeks later. Intensive treatment with topical amikacin (25 mg/mL) and ciprofloxacin and partial excision of the flap led to resolution of the infection after 2 months of therapy. In addition, topical methylprednisolone (1 mg/mL) was necessary to control inflammation and pain. No recurrence was found during the 1-year follow-up period. The final uncorrected and corrected visual acuity in the right eye was 20/25 and 20/20, respectively. CONCLUSION: Mycobacterium keratitis is a potentially vision-threatening complication after LASIK. Early recognition of the organisms and aggressive treatment if infection develops may improve the outcome. When surgery is indicated, partial flap amputation, which may help to resolve the infection and preserve better vision, may be considered. PMID- 15778612 TI - Spontaneous perforation of the cornea following silicone oil keratopathy. AB - PURPOSE: To report a sight-threatening complication of silicone oil injection for management of complex retinal detachment. METHODS: Review of case record. RESULTS: Spontaneous perforation of cornea may occur following silicone oil injection into an eye for complex retinal detachments. Risk factors are longer duration of oil in the eye, aphakia, silicone oil in the anterior chamber, and extensive and multiple surgeries. CONCLUSIONS: In high-risk cases, regular assessment of corneal status and removal of silicone oil may help to prevent such a complication. PMID- 15778613 TI - Features of the iridocorneal endothelial syndrome on confocal microscopy. AB - PURPOSE: To present a subtle case of iridocorneal endothelial (ICE) syndrome and discuss the utility of in vivo confocal microscopy in the evaluation of this disorder. Previous reports of the confocal microscopic features of ICE syndrome are reviewed. METHODS: A 32-year-old man presented with decreased vision and halos. Slit-lamp biomicroscopy and in vivo confocal microscopy were used to evaluate the etiology of his complaints. Clinical photographs and confocal micrographs were analyzed. RESULTS: In vivo confocal microscopy revealed marked asymmetry between the right and left corneal endothelial layers with pleomorphic epithelioid cells on the right, some with hyperreflective nuclei. A transition between cells with uniform appearance and dark nuclei and a highly irregular cellular arrangement with hyperreflective nuclei was present. CONCLUSIONS: This report supports the clinical utility of in vivo confocal microscopy in the evaluation of ICE syndrome. It demonstrates that the histopathologic features of ICE syndrome on scanning electron microscopy may be shown clinically using high resolution confocal microscopy. PMID- 15778614 TI - Conjunctival melanoma arising from diffuse primary acquired melanosis in a young black woman. AB - PURPOSE: To report a case of conjunctival melanoma arising from diffuse primary acquired melanosis (PAM) with atypia in a young black woman in the context of previously published cases of this lesion in blacks. METHODS: Retrospective case report with literature review. The number and percentage of conjunctival melanomas occurring in black patients were determined from case series in which race was specified, published from 1950 to the present. RESULTS: Nodular multifocal conjunctival melanoma in a 30-year-old black woman was treated using surgical excision and adjuvant cryotherapy. Extensive PAM with severe atypia, including areas of microinvasive melanoma, was treated using topical mitomycin C. Literature review revealed 35 cases of conjunctival melanoma occurring in black patients. No previous reports of mitomycin C use in black patients with melanoma or PAM were identified. CONCLUSIONS: Conjunctival melanoma is an exceedingly rare tumor in black patients. The current case brings the total of reported cases to 36. We successfully treated nodular melanoma and diffuse PAM in a young black woman using a combination of excision with cryotherapy and topical mitomycin C, suggesting that these lesions are amenable to the same types of therapy previously described for white patients. PMID- 15778615 TI - Mycobacterium chelonae keratitis: elucidation of diagnosis through evaluation of smears of fluid from patient's contact lens care system. AB - PURPOSE: To describe the microbiologic diagnosis of putative Mycobacterium chelonae keratitis in a soft contact lens wearer by initial evaluation of Gram- and Kinyoun (acid fast)-stained smears of fluid from patient's contact lens care system. METHODS: Corneal ulcer of suspected Acanthamoeba etiology developed in a 28-year-old soft contact lens wearer. After corneal scrapings were negative, microbiologic consultation led to evaluation of stained smears and culture of fluid from patient's contact lens care system. RESULTS: Gram stain of smears showed a polymicrobic flora distinguished by numerous gram-positive, beaded, tightly banded, "diphtheroid-like" bacilli strongly suggestive of a rapidly growing mycobacterial species. Kinyoun-stained smears revealed innumerable acid fast bacilli singly and in tightly woven bundles (cords), which culturally proved to be M. chelonae. Treatment with ciprofloxacin and amikacin resolved the ulcer. CONCLUSION: Diagnosis of M. chelonae keratitis in contact lens wearers is often delayed or even overlooked. Additionally, in the absence of overt corneal injury, eg, trauma or surgery, a source for the infecting mycobacterial species in the setting of contact lens wear has not been identified. If searched for, however, as in the present case, the patient's contact lens care system may serve as the reservoir for the microorganism. Staining for acid-fast bacilli is further recommended when smears of contact lens care solution of a patient with a corneal ulcer shows the presence of gram-positive "diphtheroid-like" organisms. PMID- 15778616 TI - Detection of corneal epithelial defect through amniotic membrane patch by fluorescein. AB - PURPOSE: To demonstrate the usefulness of fluorescein dye to detect a corneal epithelial defect through an amniotic membrane patch in 4 cases. PATIENTS AND METHOD: Two patients with painful bullous keratopathy, 1 patient with total limbal deficiency, and 1 patient with acute chemical burn underwent surgeries in conjunction with amniotic membrane as patch. Postoperatively, slit-lamp biomicroscopic examination with and without fluorescein staining was performed. RESULTS: The status of epithelialization, ie, corneal epithelial defect on either denuded corneal stroma or amniotic membrane graft, was indiscernible by slit-lamp biomicroscopy through the amniotic membrane patch. Nevertheless, it could be detected by fluorescein staining, especially after saline rinse, in all 4 cases. CONCLUSIONS: Fluorescein staining permits clinicians to avoid prematurely removing the amniotic membrane patch during the treatment of a persistent corneal epithelial defect. PMID- 15778617 TI - Epibulbar nodular fasciitis associated with floppy eyelids. AB - PURPOSE: To report a case of epibulbar nodular fasciitis in a patient with floppy eyelids. METHOD: Observational case report. Clinical, histologic, and immunohistochemical findings in a patient who underwent excision of a subconjunctival mass. RESULTS: The clinical examination revealed floppy eyelids and an epibulbar mass in the right eye. The histopathology from an excisional biopsy of the mass was consistent with nodular fasciitis. CONCLUSIONS: Nodular fasciitis is a rare, benign, soft-tissue tumor that has not been previously reported in association with floppy eyelids. The cause of both disorders is thought to be repetitive trauma, suggesting that eye rubbing may be a common underlying etiology. PMID- 15778618 TI - How to best treat adenoviral corneal opacities. PMID- 15778620 TI - Fixed dilated pupil (Urrets-Zavalia syndrome) in corneal dystrophies. PMID- 15778621 TI - Effect of pterygium surgery on corneal topography. PMID- 15778623 TI - From the editor. PMID- 15778625 TI - A panel study of physical activity in Taiwanese youth: testing the revised health promotion model. AB - The rate of inactivity among young people in Taiwan remains high. However, few studies have explored physical activity patterns with a longitudinal approach in this population. Based on the revised health-promotion model, this study used 2 wave panel data to test a structural model of how individual characteristics, cognitions, and interpersonal influences predicted physical activity of Taiwanese adolescents. The findings suggest that gender, social support, modeling, self efficacy, and perceived benefits and barriers to performing physical activity directly and indirectly influence the behavior of physical activity in Taiwanese adolescents. The hypothesized model explained 25% of the variance in physical activity. The implications for parents and policy are discussed. PMID- 15778626 TI - Physical activity in physical education: teacher or technology effects. AB - This study assessed if wearing a heart rate monitor influenced student activity levels in elementary physical education. Data were analyzed for 4 students using an alternating treatment design to assess differential effects between accelerometer activity counts obtained from students when wearing the heart rate monitor and when they were not wearing the monitor. Results show that (a) there was no difference in activity counts between the 2 conditions, (b) boys had higher means than girls, and (c) the variance between more and less active boys was greater than the variance among the girls. PMID- 15778627 TI - Perceptions of health status and play activities in parents of overweight Hispanic toddlers and preschoolers. AB - Childhood overweight among lower socioeconomic, Hispanic children has increased. Interviews regarding health status and play patterns were conducted with 76 predominantly Hispanic mothers of overweight toddlers and preschoolers served by Women, Infants, and Children (WIC). Most participants believed their child was healthy and half were unconcerned about their child's weight. Most parents reported having a safe place to play and access to a playground, although gender differences were found. Access to an outside play area was related to amount of active play activities. Children watched an average of 1.7 hours per day of television. Health professionals must partner with parents to address childhood obesity. PMID- 15778628 TI - Keeping all children healthy: challenges to leading an active lifestyle for preschool children qualifying for at-risk programs. AB - This study examined contextual factors associated with physical activity of urban African American preschoolers (N = 59). Qualitative research methodologies utilized data from home visits, caregiver and child interviews, field notes, and document collection. The primary finding suggested, "In underserved communities fundamental barriers exist that obstruct young children's ability to be physically active." Four themes were developed revealing that outside environments limited opportunities for physical activity, home environments resulted in sedentary behaviors, and communities had limited physical activity role models. Despite this, young children enjoyed being physically active. PMID- 15778629 TI - Health inequity: the plight of uninsured children in a rural Alabama county and the plan to cure it. AB - Many children in the United States do not have access to health insurance. Providing health insurance for children has been particularly challenging in rural America. This article describes and evaluates a local plan to provide access to health care for school children in a rural Alabama county. A triangulated methodology (personal interviews, ad hoc survey focus groups, US census and health fair data) was used in the evaluation. Gains were made in enrolling children despite some limitations, especially in rural outreach. The most successful aspect of the program was a partnership between local leaders, health providers, and educators to provide impetus for a coordinated plan. The stability of the program is uncertain because of diminishing resources and the negative effects of economic recession. PMID- 15778630 TI - The independent impact of household- and neighborhood-based social determinants on early childhood caries: a cross-sectional study of inner-city children. AB - This study examined the relationship between early childhood caries (ECC) and measures of socioeconomic status as environmental determinants of oral health inequalities. Dental caries and quality of oral hygiene were reported on 384 young inner-city children (24-35 months) by clinical examination. Early childhood caries was diagnosed in 18.5% of children. Ethnicity (P < 0.001) and neighborhood (P < 0.001) were the social variables significantly predictive of ECC in a logistic regression model. Living in a deprived neighborhood and the mother being of East European descent were the social variables with strongest association with ECC. These variables can be considered adequate criteria to identify groups at risk within the overall population. PMID- 15778631 TI - The health of chronically ill children: lessons learned from assessing family caregiver quality of life. AB - This study aims to assess family caregiver quality of life (QOL) and its relationship to the health of chronically ill children. Family caregivers (n = 100) completed the Medical Outcomes Short Form 36, Beck Depression Inventory, and Caregiver Quality of Life for Cystic Fibrosis Scale. Disease severity data were collected for children with cystic fibrosis. Analysis revealed significant QOL issues; and as the child's disease severity increases, the QOL of the caregiver decreases. A relationship exists between the child's and caregiver's health. Program recommendations to benefit family caregiver health and in turn benefit health of chronically ill children are discussed. PMID- 15778632 TI - Building holistic HIV/AIDS responses in African American urban faith communities: a qualitative, multiple case study analysis. AB - Holistic prevention strategies are increasingly more effective in eradicating the national human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) health crisis, which disproportionately affects African Americans. Faith communities have been integral in advancing African American community welfare; however, little is understood about their evolving role in HIV prevention. This article reports the findings from a study conducted in Washington, DC, that identifies the factors that shape the holistic development of HIV/AIDS-prevention programs within African American faith communities. By providing policy recommendations, the research illuminated a useful theoretic framework and opportunities to more holistically address current social and structural challenges in prevention efforts among faith-health leaders in similar environments. PMID- 15778633 TI - Integrating research on racial and ethnic disparities in health care over place and time. PMID- 15778634 TI - Hospital-level racial disparities in acute myocardial infarction treatment and outcomes. AB - BACKGROUND: Previous studies have documented racial disparities in treatment of acute myocardial infarction (AMI) among Medicare beneficiaries. However, the extent to which unobserved differences between hospitals explains some of these differences is unknown. OBJECTIVE: The objective of this study was to determine whether the observed racial treatment disparities for AMI narrow when analyses account for differences in where blacks and whites are hospitalized. RESEARCH DESIGN: Retrospective observational cohort study using Medicare claims and medical record review. SUBJECTS: This study included 130,709 white and 8286 black Medicare patients treated in 4690 hospitals in 50 US states for confirmed AMI in 1994 and 1995. MEASURES: Measures in this study were receipt of reperfusion, aspirin, and smoking cessation counseling during hospitalization; prescription of aspirin, angiotensin-converting enzyme inhibitor, and beta-blocker at hospital discharge; receipt of cardiac catheterization, percutaneous coronary intervention (PCI), or bypass surgery (CABG) within 30 days of AMI; and 30-day and 1-year mortality. RESULTS: Within-hospital analyses narrowed or erased black-white disparities for medical treatments received during the acute hospitalization, widened black-white disparities for follow-up surgical treatments, and augmented the survival advantage among blacks. These findings indicate that, on average, blacks went to hospitals that had lower rates of evidence-based medical treatments, higher rates of cardiac procedures, and worse risk-adjusted mortality after AMI. CONCLUSIONS: Incorporating the hospital effect altered the findings of racial disparity analyses in AMI and explained more of the disparities than race. A policy of targeted hospital-level interventions may be required for success of national efforts to reduce disparities. PMID- 15778635 TI - Technology diffusion, hospital variation, and racial disparities among elderly Medicare beneficiaries: 1989-2000. AB - BACKGROUND: Low rates of technology utilization in hospitals with high proportions of black inpatients may be a remediable cause of healthcare disparities. OBJECTIVES: Our objective was to determine how differences in technology utilization among hospitals contributed to racial disparity and if temporal reduction in hospital procedure rate variation resulted in decreased racial disparity for these technologies. METHODS: We identified 2,348,952 elderly Medicare beneficiaries potentially eligible for 1 of 5 emerging medical technologies from 1989-2000 and determined if these patients had received the indicated procedure within 90 days of their qualifying hospital admission. Initial multivariate regression models adjusted for age, race, sex, admission year, clinical comorbidity, community levels of education and income, and academic/urban hospital admission. The inpatient racial composition of each patient's admitting hospital and time-race interactions were added as covariates to subsequent models. RESULTS: Blacks had significantly lower adjusted rates (P < 0.001) compared with whites for tissue replacement of the aortic valve, internal mammary artery coronary bypass grafting, dual-chambered pacemaker implantation, and lumbar spinal fusion. Hospitals with > 20% black inpatients were less likely to perform these procedures on both white and black patients than hospitals with < 9% black inpatients, and racial disparity was greater in hospitals with larger black populations. There were no temporal reductions in racial disparities. CONCLUSIONS: Blacks may be disadvantaged in access to new procedures by receiving care at hospitals that have both lower procedure rates and greater racial disparity. Policies designed to ameliorate racial disparities in health care must address hospital variation in the provision of care. PMID- 15778636 TI - Race/ethnic disparities in utilization of lifesaving technologies by Medicare ischemic heart disease beneficiaries. AB - OBJECTIVE: The objective of this study was to explain race/ethnic disparities in hospitalizations, utilization of high-technology diagnostic and revascularization services, and mortality of elderly ischemic heart disease (IHD) patients. DESIGN: A longitudinal Medicare claims database of all Part A hospital and Part B physician services provided elderly patients admitted for IHD in 1997 is used to construct admission, utilization, and mortality rates for whites and blacks, Asians, Hispanics, and American Indians. Z-scores are used to test differences in rates between whites and minorities at the 99% confidence level. Logistic and proportional hazard models are used to predict the likelihood of revascularization and its effects on race/ethnic survival 2 years postdischarge. SETTING: The setting of this study was an acute hospital supplemented by all ambulatory Part B outpatient providers of care. PATIENTS/PARTICIPANTS: Participants included all 700,000 age 65+ Medicare beneficiaries in fee-for service identified with IHD as a primary diagnosis on admission in 1997. MEASUREMENTS AND MAIN RESULTS: Whites were 26% more likely to be admitted for IHD than blacks, 50% more likely than Asians, 5% more than American Indians, but 3% less likely than Hispanics. Once admitted, elderly blacks and American Indians undergo invasive diagnostic and surgical revascularization far less often than whites (P < 0.01), although blacks are equally as likely as whites to be admitted to an open heart hospital. Controlling for other factors, whites reduce their 2 year mortality by 20% by undergoing revascularization 41% of the time. Blacks gain only 11% as a result of much lower rates and gains to revascularization than whites. Asians and Hispanics were slightly more likely than whites to undergo revascularization but gain less than whites from the procedure. CONCLUSIONS: Despite having similar Medicare health insurance coverage, elderly utilization and IHD mortality rates differ markedly not only between whites and minorities, but within minority groups themselves. A large, nationally representative survey of physicians and patients is needed to distinguish between systemwide "failures to refer" and patient "aversions to surgery" as explanations for lower black rates of surgical interventions. PMID- 15778637 TI - Competition and health plan performance: evidence from health maintenance organization insurance markets. AB - OBJECTIVE: We sought to assess whether health maintenance organizations (HMOs) operating in competitive markets, or markets with substantial HMO penetration, perform better on the standardized Health Plan Employer Data and Information Set (HEDIS) and Consumer Assessment of Health Plans Survey (CAHPS) measures. STUDY DESIGN: We performed a secondary analysis of nonexperimental, cross-sectional data. DATA SOURCES: Data were obtained from a variety of sources, including the National Committee for Quality Assurance (NCQA), Interstudy, the Area Resource File, the U.S. Office of Personnel Management, and the U.S. Department of Labor. METHODS: Multiple Indicator Multiple Cause models were used to simultaneously estimate 6 latent quality variables from 35 HEDIS and CAHPS measures and to relate these latent variables to HMO competition and HMO penetration while controlling for other health plan and market characteristics. PRINCIPAL FINDINGS: Greater competition, as measured by the Herfindahl index, was associated with inferior health plan performance on 3 of 6 quality dimensions. Plans in markets with greater HMO penetration perform better on HEDIS- but not CAHPS-based dimensions of performance. Plans that make their data available publicly perform significantly better on both the HEDIS and CAHPS domains, performing one third to three quarters of a standard deviation better than plans that don't make their results available publicly. CONCLUSIONS: Plans in more competitive markets in 1999 did not achieve better quality after controlling for other important covariates, although plans in markets with a high degree of HMO penetration are performing better on the HEDIS quality dimensions. Although our study design cannot determine causality, the results suggest reason to revisit the belief that competition among HMOs will inherently improve quality. PMID- 15778638 TI - Utilities for prostate cancer health states in men aged 60 and older. AB - PURPOSE: We sought to measure utilities for prostate cancer health states in older men. METHODS: A total of 162 men aged 60 years or older (52% of whom had been diagnosed with prostate cancer) provided standard gamble utilities for 19 health states associated with prostate cancer or its treatment using an interactive, computer-based utility assessment program. Demographics and experience with specific health states were examined as predictors of ratings using ordinary least squares regression analysis. RESULTS: Mean utilities ranged from 0.67 to 0.84 for living with symptom-free cancer under conservative management ("watchful waiting") and from 0.71 to 0.89 for symptoms occurring with treatment (prostatectomy, radiation, and hormone ablation). For long-term treatment complications, bowel problems (0.71) were rated as significantly worse than impotence (0.89), urinary difficulty (0.88), or urinary incontinence (0.83). Combinations of these conditions were rated as significantly worse than individual component states. Men who had experienced impotence or urinary incontinence rated these states as slightly better than men who had not experienced the specific problems. CONCLUSIONS: Both "watchful waiting" and treatment complications from prostate cancer treatments can have large impacts on quality of life. Mean ratings are important for use in policy-making and cost effectiveness analyses. Variation in ratings across patients suggests that mean scores do not reflect individual preferences and that shared decision-making may be best for clinical decisions. PMID- 15778639 TI - Cultural competence: a systematic review of health care provider educational interventions. AB - OBJECTIVE: We sought to synthesize the findings of studies evaluating interventions to improve the cultural competence of health professionals. DESIGN: This was a systematic literature review and analysis. METHODS: We performed electronic and hand searches from 1980 through June 2003 to identify studies that evaluated interventions designed to improve the cultural competence of health professionals. We abstracted and synthesized data from studies that had both a before- and an after-intervention evaluation or had a control group for comparison and graded the strength of the evidence as excellent, good, fair, or poor using predetermined criteria. MAIN OUTCOME MEASURES: We sought evidence of the effectiveness and costs of cultural competence training of health professionals. RESULTS: Thirty-four studies were included in our review. There is excellent evidence that cultural competence training improves the knowledge of health professionals (17 of 19 studies demonstrated a beneficial effect), and good evidence that cultural competence training improves the attitudes and skills of health professionals (21 of 25 studies evaluating attitudes demonstrated a beneficial effect and 14 of 14 studies evaluating skills demonstrated a beneficial effect). There is good evidence that cultural competence training impacts patient satisfaction (3 of 3 studies demonstrated a beneficial effect), poor evidence that cultural competence training impacts patient adherence (although the one study designed to do this demonstrated a beneficial effect), and no studies that have evaluated patient health status outcomes. There is poor evidence to determine the costs of cultural competence training (5 studies included incomplete estimates of costs). CONCLUSIONS: Cultural competence training shows promise as a strategy for improving the knowledge, attitudes, and skills of health professionals. However, evidence that it improves patient adherence to therapy, health outcomes, and equity of services across racial and ethnic groups is lacking. Future research should focus on these outcomes and should determine which teaching methods and content are most effective. PMID- 15778640 TI - Nonresponse bias in a survey of patient perceptions of hospital care. AB - BACKGROUND: Incomplete participation is of particular concern for surveys of patient perceptions of care because patients who have negative opinions may be least likely to participate. OBJECTIVE: We sought to examine indirect evidence of nonresponse bias. DESIGN: We re-analyzed data from a cross-sectional patient survey. SUBJECTS: Our subjects were patients discharged from a Swiss hospital (n = 2156). MEASURES: We measured the following: (1) an observed problem score, based on 15 key items of the Picker Patient Experience questionnaire, (2) a predicted problem score, and (3) a participation propensity score. The latter scores were computed for all eligible patients, including those who did not return the survey, from routinely available baseline data. RESULTS: The participation rate was 70% (n = 1518), and the mean problem score was 29.9 (SD 23.8). Early respondents reported significantly fewer problems than late respondents (28.6 versus 32.9, P = 0.001). Participation propensity scores were progressively lower in early respondents (mean 74.2), late respondents (70.7), and nonrespondents (63.9, P < 0.001); the pattern was similar for predicted problem scores (early respondents: 29.5; late respondents: 30.5; nonrespondents: 33.4, P < 0.001). The propensity to participate was negatively associated with the problem score (Pearson r = -0.19). Finally, predictors of participation were similar to predictors of problem scores. CONCLUSIONS: The tendency to participate in the survey was negatively associated with the report of problems during hospitalization. Nevertheless, increasing participation from 30% to 70% had only a modest influence on the final conclusions of the survey. PMID- 15778641 TI - Improving depression care for older, minority patients in primary care. AB - OBJECTIVE: Few older minorities receive adequate treatment of depression in primary care. This study examines whether a collaborative care model for depression in primary care is as effective in older minorities as it is in nonminority elderly patients in improving depression treatment and outcomes. STUDY DESIGN: A multisite randomized clinical trial of 1801 older adults comparing collaborative care for depression with treatment as usual in primary care. Twelve percent of the sample were black (n = 222), 8% were Latino (n = 138), and 3% (n = 53) were from other minority groups. We compared the 3 largest ethnic groups (non-Latino white, black, and Latino) on depression severity, quality of life, and mental health service use at baseline, 3, 6, and 12 months after randomization to collaborative care or usual care. PRINCIPAL FINDINGS: Compared with care as usual, collaborative care significantly improved rates and outcomes of depression care in older adults from ethnic minority groups and in older whites. At 12 months, intervention patients from ethnic minorities (blacks and Latinos) had significantly greater rates of depression care for both antidepressant medication and psychotherapy, lower depression severity, and less health-related functional impairment than usual care participants (64%, 95% confidence interval [CI] 55-72 versus 45%, CI 36-55, P = 0.003 for antidepressant medication; 37%, CI 28-47 versus 13%, CI 6-19, P = 0.002 for psychotherapy; mean = 0.9, CI 0.8-1.1 versus mean = 1.4, CI 1.3-1.5, P < 0.001 for depression severity, range 0-4; mean = 3.7, CI 3.2-4.1, versus mean = 4.7, CI 4.3-5.1, P < 0.0001 for functional impairment, range 0-10). CONCLUSIONS: Collaborative Care is significantly more effective than usual care for depressed older adults, regardless of their ethnicity. Intervention effects in ethnic minority participants were similar to those observed in whites. PMID- 15778642 TI - Do patients consistently report comorbid conditions over time?: results from the prostate cancer outcomes study. AB - BACKGROUND: Comorbidity is an important dimension of patient health status. However, limited attention has been given to assessing the reliability of patient reported data on comorbid conditions. OBJECTIVES: We sought to evaluate the consistency of self-report of 12 comorbid conditions at 3 distinct time points and examine the sociodemographic, clinical, health status, and survey factors associated with reliability. STUDY DESIGN: We undertook a longitudinal cohort analysis of survey and medical record abstract data obtained from a population based sample of 3095 prostate cancer survivors. METHODS: Consistent and inconsistent response patterns were assessed using descriptive statistics and regression modeling. RESULTS: More than half of the cohort demonstrated consistent responses to all comorbid condition questionnaire items. Arthritis had the highest (13%) and liver disease the lowest (1%) proportion of inconsistent responses. Older age, lower income, and poorer general and mental health status were significant predictors of inconsistent responses. Subset analyses of the 4 most prevalent comorbid conditions (ie, arthritis, diabetes, hypertension, and depression) showed that respondents who reported that they were limited by, or taking prescription medicine for, the condition were more likely to provide consistent responses than those neither limited nor taking medicine. Response consistencies of 92% or better were obtained for 11 of the 12 conditions. CONCLUSIONS: Men with a relatively recent prostate cancer diagnosis are generally able to provide reliable reports of their concomitant health conditions. To increase the likelihood of obtaining reliable data, investigators should consider ascertaining condition severity and current medical management when querying subjects about comorbid conditions in surveys. PMID- 15778644 TI - Increasing response rates in a survey of Medicaid enrollees: the effect of a prepaid monetary incentive and mixed modes (mail and telephone). AB - OBJECTIVES: We sought to evaluate the effect of pairing a mixed-mode mail and telephone methodology with a prepaid US 2.00 dollars cash incentive on response rates in a survey of Medicaid enrollees stratified by race and ethnicity. RESEARCH DESIGN: Sampling was conducted in 2 stages. The first stage consisted of a simple random sample (SRS) of Medicaid enrollees. In the second stage, American Indian, African American, Latino, Hmong, and Somali enrollees were randomly sampled. A total of 8412 enrollees were assigned randomly to receive a mail survey with no incentive or a US 2.00 dollars bill. RESULTS: The response rate within the SRS after the mail portion was 54% in the incentive group and 45% in the nonincentive group. Response rates increased considerably with telephone follow-ups. The incentive SRS response rate increased to 69%, and the nonincentive response rate increased to 64%. Differences between incentive conditions are more pronounced after the first mailing (P < 0.01); almost all differences remained significant (P < 0.05) after the completion of the mail mode. The inclusion of the US 2.00 dollars incentive had similar effects on response rates and cost across the different racial and ethnic strata, except for Latino enrollees. CONCLUSIONS: A mixed-mode mail and telephone methodology is effective for increasing response rates in a Medicaid population overall and within different racial and ethnic groupings. The effectiveness of this strategy can be enhanced, in terms of response rate and cost, by including a US 2.00 dollars prepaid incentive. PMID- 15778643 TI - Insurance coverage, usual source of care, and receipt of clinically indicated care for comorbid conditions among adults living with human immunodeficiency virus. AB - BACKGROUND AND OBJECTIVES: Associations of insurance coverage and source of care with use of human immunodeficiency virus (HIV)-related health, mental health, and substance abuse services are examined in a large, diverse, highly active antiretroviral therapy-era cohort. METHODS: Adults who were infected with HIV (n = 3818) were interviewed in clinics and community agencies in Los Angeles, Milwaukee, New York, and San Francisco regarding drug use behaviors, health status, and health care utilization. RESULTS: Most participants were insured by Medicaid. During the previous 3 months, 90% of privately insured, 87% of publicly insured, and 78% of uninsured participants had visited any provider. Publicly and privately insured participants were similar in receipt of antiretrovirals, prophylaxis against Pneumocystis carinii pneumonia, substance abuse services, and antidepressants. Uninsured participants were less likely to receive antiretrovirals but were more likely to use substance abuse services. Participants with no usual source of care were less likely to receive PCP prophylaxis. CONCLUSIONS: A lack of insurance is associated with barriers to care, but the advantage of private over public coverage appears smaller than in previous studies. PCP prophylaxis, substance abuse treatment, and antidepressants remain markedly underutilized. Educational initiatives about these treatments targeting providers and patients are indicated. PMID- 15778645 TI - Free internet access, the digital divide, and health information. AB - BACKGROUND: The Internet has emerged as a valuable tool for health information. Half of the U.S. population lacked Internet access in 2001, creating concerns about those without access. Starting in 1999, a survey firm randomly invited individuals to join their research panel in return for free Internet access. This provides a unique setting to study the ways that people who had not previously obtained Internet access use the Internet when it becomes available to them. METHODS: In 2001-2002, we surveyed 12,878 individuals 21 years of age and older on the research panel regarding use of the Internet for health; 8935 (69%) responded. We analyzed respondents who had no prior Internet access, and then compared this group to those who had prior Internet access. RESULTS: Among those newly provided free Internet access, 24% had used the Internet for health information in the past year, and users reported notable benefits, such as improved knowledge and self-care abilities. Not surprisingly, the no-prior Internet group reported lower rates of using the Internet (24%) than the group that had obtained Internet access prior to joining the research panel (40%), but the 2 groups reported similar perceptions of the Internet and self-reported effects. CONCLUSIONS: Those who obtained Internet access for the first time by joining the panel used the Internet for health and appeared to benefit from it. Access helps explain the digital divide, although most people given free access do not use the Internet for health information. PMID- 15778647 TI - Response to an editorial in the January 2005 issue of Medical Care. PMID- 15778648 TI - Read fiction to know nursing. PMID- 15778649 TI - The impact of hospital nursing characteristics on 30-day mortality. AB - BACKGROUND: Evidence indicates that hospital nursing characteristics such as staffing contribute to patient outcomes. Less attention has been given to other hospital nursing characteristics central to optimal professional practice, namely nurse education and skill mix, continuity of care, and quality of the work environment. OBJECTIVE: To assess the relative effects and importance of nurse education and skill mix, continuity of care, and quality of work environment in predicting 30-day mortality after adjusting for institutional factors and individual patients characteristics. METHOD: A cross-sectional analysis of outcome data for 18,142 patients discharged from 49 acute care hospitals in Alberta, Canada, for diagnoses of acute myocardial infarction, congestive heart failure, chronic obstructive pulmonary disease, pneumonia, or stroke between April 1, 1998, and March 31, 1999, was done. Mortality data were linked to patient demographic and comorbidity factors, institutional characteristics, and hospital nursing characteristics derived from a survey of all registered nurses working in acute care hospitals. RESULTS: Using multilevel analysis, it was determined that the log-odds for 30-day mortality varied significantly across hospitals (variance = .044, p < .001). Patient comorbidities and age explained 44.2% of the variance in 30-day mortality. After adjustment for patient comorbidities and demographic factors, and the size, teaching, and urban status of the study hospitals in a fixed-effects model, the odds ratios (95% confidence interval) of the significant hospital nursing characteristics that predict 30-day mortality were as follows: 0.81 (0.68-0.96) for higher nurse education level, 0.83 (0.73-0.96) for richer nurse skill mix, 1.26 (1.09-1.47) for higher proportion of casual or temporary positions, and 0.74 (0.60-0.91) for greater nurse-physician relationships. The institutional and hospital nursing characteristics explained an additional 36.9%. DISCUSSION: Hospital nursing characteristics are an important consideration in efforts to reduce the risk of 30-day mortality of patients. PMID- 15778650 TI - Structural model for quality of life of patients with chronic cardiovascular disease in Korea. AB - BACKGROUND: Health promotion among those with chronic illness has been proposed as a strategy to contain health care cost and enhance quality of life (QOL), but little research has been done in Korea on QOL focused on health promotion of patients with chronic cardiovascular disease (CCVD). OBJECTIVES: This study was designed to estimate a structural model to represent the QOL of patients with CCVD, on the basis of a literature review and Pender's health promotion model (HPM). METHODS: Data were collected by questionnaire from 436 patients with CCVD in a university hospital in Seoul from August 2001 to July 2002. Data analysis included descriptive statistics and covariance structural analysis. RESULTS: As the fit of the hypothetical model to the data was only moderate, it was modified by excluding 10 paths and including free parameters. The modified model with path showed a good fit to the empirical data (chi = 508.12, p = .001, GFI = .95, AGFI = .93, NFI = .98, NNFI = .98, RMSEA = .04). Health-promoting behavior (HPB) and self-efficacy were found to have a significant direct effect on QOL. Health perception, self-esteem, perceived barriers to action, and preference were found to have indirect effects on QOL. The variables of this model explain 63% of the variance in QOL. DISCUSSION: The modified model is considered appropriate to explain and predict QOL of patients with CCVD. Therefore, it can effectively be used as a reference model for further studies and to suggest directions in nursing practice. PMID- 15778651 TI - Effectiveness of a behavioral change intervention in Thai elders after knee replacement. AB - BACKGROUND: After total knee replacement, elders need an effective intervention to change exercise and physical activity behavior. OBJECTIVES: This study examined the effects of a behavioral change intervention (BCI) on self-efficacy and outcome expectations for exercise and functional activity, physical activity participation, and physical performance of older adults. METHODS: The study was based on the social cognitive theory (SCT), with a longitudinal quasi experimental, pretest-posttest control group design. Sixty-three Thai elders undergoing knee replacement surgery were studied. The experimental group received a BCI based on SCT given by the investigator. Outcome measures were the Self Efficacy for Exercise Scale (SEES) and Self-Efficacy for Functional Activity Scale (SEFAS), the Outcome Expectations for Exercise Scale (OEES) and Outcome Expectations for Functional Activity Scale (OEFAS), the Physical Performance Test (PPT), and the Physical Activity Diary (PAD). RESULTS: The experimental group had significantly greater improvements in self-efficacy for exercise, outcome expectations for exercise, and functional activity, significantly more participation in exercise and walking, and significantly greater improvement in physical performance than did the control group at postoperative weeks 2 and 6. DISCUSSION: The BCI based on SCT was effective in changing the outcomes in the expected direction. This BCI may be applicable, with modifications, to elders in other situations in which changing behavior is the key to recovery. PMID- 15778652 TI - A randomized clinical trial of an HIV-risk-reduction intervention among low income Latina women. AB - BACKGROUND: HIV infection has increased within the Latina community more than in any other ethnic or racial group within the United States. Latinas comprise only 13% of the U.S. population, yet they account for 20% of the cumulative reported cases of AIDS. OBJECTIVES: The purpose of the study was to evaluate a randomized culturally tailored intervention to prevent high-HIV-risk sexual behaviors for Latina women residing in urban areas. METHODS: Mexican and Puerto Rican women (18 44 years of age; N = 657) who were sexually active during the previous 3 months were recruited and randomized into intervention and control groups. The intervention, facilitated by bilingual, bicultural, trained Latina women, consisted of culturally tailored sessions on understanding their bodies, HIV/AIDS and sexually transmitted diseases, condoms (myths and use), negotiating safer sex practices, violence prevention, and partner communication. Bivariate and multivariate analyses assessed changes from baseline. RESULTS: The intervention improved HIV knowledge, partner communication, risk-reduction behavioral intentions, and condom use, and decreased perceived barriers to condom use. DISCUSSION: The efficacy of a culturally-sensitive intervention to reduce HIV/AIDS-risk behaviors in Latina women was demonstrated in the current study. PMID- 15778653 TI - Reducing assaults against nursing home caregivers. AB - BACKGROUND: Nursing assistants (NAs) working in long-term care have the highest incidence of workplace assault among all workers in the United States. OBJECTIVES: The objective of this study was to test the effectiveness of a violence-prevention intervention, on the basis of Social Cognitive Theory to increase knowledge, self-efficacy, and skills, and to decrease assaults. METHODS: Investigators conducted a quasi-experimental study with 138 NAs in 3 intervention and 3 comparison homes. A baseline questionnaire was used to obtain information on demographics, employment, and violence experience. At pre-, post- and 6 months after the intervention, all participants completed the State Trait Anger Inventory and the Knowledge and Self-Efficacy Survey, carried an Assault log for 80 hr of work, and participated in a simulation exercise to assess violence prevention skills. Tabulations, analysis of variance, and Poisson regression were used to analyze the data. RESULTS: The intervention participants showed significant increases in knowledge, self-efficacy, and violence-prevention skills. Although the intervention had no significant main effect on the incidence of assaults, there was an interaction effect between the intervention and the number of preintervention assaults. The intervention had a significant effect on those NAs who had fewer than 6 assaults preintervention (p < .001) and no significant effect on those who had more than 7 assaults on preintervention. There were significant relationships between assaults and the following covariates: age, state anger, and the number of residents assigned. DISCUSSION: Although the incidence of violence cannot be eliminated, it can be decreased and it should never be tolerated or accepted as "part of the job" because to do so devalues the NA. PMID- 15778654 TI - The Consolidated Standards of Reporting Trials (CONSORT): Guidelines for reporting randomized trials. AB - BACKGROUND: The rules for reporting the results of randomized controlled trials (RCTs) in published manuscripts have been elusive, and many nurse scientists have learned to write research reports by trial and error. APPROACH: New comprehensive guidelines for reporting RCTs are now available to authors, editors, and readers, a development that is likely to improve the quality of published reports. The Consolidated Standards of Reporting Trials (CONSORT) guidelines consist of two parts, a 22-item checklist and a flowchart template, that track the flow of all participants through the RCT. Currently, some journals, including Nursing Research, have adopted CONSORT and suggest or require its use in submitted manuscripts, but other journals do not. RESULTS: The CONSORT has potential to be useful beyond writing manuscripts. Because CONSORT is a comprehensive guide to essential elements of an RCT report, it also provides a useful guide for designing RCTs and writing funding proposals that include all the essential elements of a trial. It also could be a useful synopsis of important concepts when teaching research design to doctoral students. DISCUSSION: Reports of RCTs that use CONSORT are easier to read, interpret, and evaluate for relevancy to clinical practice. In addition, excellent reports of research increase the visibility of nursing research beyond our discipline. The CONSORT adds value to nursing research because researchers and readers alike will benefit if it is used more often. PMID- 15778655 TI - Logistic regression and Bayesian networks to study outcomes using large data sets. AB - BACKGROUND: In nursing research, the interest in using large health care databases to predict nursing sensitive outcomes is growing rapidly. Traditionally, one of the most frequently used methods is logistic regression (LR), which, although powerful and familiar, has several limitations when used in the analysis of large databases. As a result, innovative approaches are required. APPROACH: To (a) introduce an innovative/alternative data analysis approach (Bayesian network), (b) discuss the constraints of LR and the complementary advantages of Bayesian networks (BNs) in working with large and multidimensional health care data, and (c) provide a fundamental understanding of the use of BNs in the nursing/health care domain. RESULTS: Studies have shown that BNs have several advantages over LR in analyzing complex and large data: (a) statistical assumptions, such as linearity and additivity, are relaxed; (b) handling of a larger number of predictors and identification of interactions among predictors is less complex; and (c) the discovery of structure, pattern, and knowledge, for example, of unknown, complex, and nonlinear relationships, in data is facilitated. CONCLUSION: Outcome studies, such as those undertaken by nurse researchers, may benefit from the examination and use of innovative approaches such as BNs to the analysis of very large and complex health care data sets. PMID- 15778656 TI - Treatment fidelity in behavior change research: a case example. AB - BACKGROUND: Treatment fidelity refers to the methodological strategies used to monitor and enhance the reliability and validity of behavioral interventions. Assuring optimal treatment fidelity also may decrease the costs of a study and help the research team explain findings. APPROACH: The Behavioral Change Consortium developed a comprehensive model of treatment fidelity that incorporates 5 areas: (a) study design, (b) training providers, (c) delivery of treatment, (d) receipt of treatment, and (e) enactment of treatment skills. The definitions of these areas and a case example (Testing the Effectiveness of the Exercise Plus Program) are provided. RESULTS: There was evidence of treatment fidelity related to delivery based on careful monitoring of the study implementation. A comprehensive plan for training of the interventionists was provided, although evidence of treatment fidelity to training was not quantified. There were evidence based on observations of treatment sessions of delivery and receipt of the intervention and evidence of enactments based on evaluation of exercise calendars. DISCUSSION: The development and implementation of a treatment fidelity plan requires a careful conceptualization of what is relevant to treatment fidelity in any given study. Monitoring of treatment fidelity ideally requires direct or indirect observations of sessions, which can be built into the study design so that costs are minimal in terms of time and resources. Monitoring treatment fidelity allows research teams to truly test interventions and to develop and implement interventions that ultimately improve the overall health and well-being of individuals. PMID- 15778658 TI - The privacy rule and the ophthalmic plastic surgeon as author. PMID- 15778659 TI - Effect of cosmetic surgery on a specialty. PMID- 15778660 TI - Conjunctival epithelial involvement by eyelid sebaceous carcinoma. The 2003 J. Howard Stokes lecture. AB - PURPOSE: To determine incidence and distribution of conjunctival epithelial involvement by eyelid sebaceous carcinoma and to make recommendations regarding its management. METHODS: The medical records were reviewed retrospectively on patients with histopathologically confirmed sebaceous carcinoma of the eyelids managed at the Oncology Service at Wills Eye Hospital. Those tumors with involvement of the conjunctival epithelium were identified and selected for further study. The incidence and distribution of epithelial involvement was determined, based on histopathology of resected tumors and map biopsies. The incidence of metastasis and tumor-related mortality was determined. Based on these findings and personal surgical experience, recommendations are made regarding management of eyelid sebaceous carcinoma with involvement of the conjunctival epithelium. RESULTS: Of 60 patients with sebaceous carcinoma, epithelial involvement of the conjunctiva was identified in 28 (47%). Of the 28 cases, the neoplasm affected the following sites: superior tarsal and fornical conjunctiva in 28 (100%), inferior tarsal conjunctiva in 19 (68%), inferior fornical conjunctiva in 18 (64%), superior bulbar conjunctiva in 19 (68%), and inferior bulbar conjunctiva in 16 (57%). The caruncle was involved in 15 (54%) and the cornea in 11 (39%). Metastasis occurred in 3 of the 28 cases (11%). Map biopsies, combined with cryotherapy, topical chemotherapy, local surgical resection, and orbital exenteration, were used to achieve local control. CONCLUSIONS: Eyelid sebaceous carcinoma was found to exhibit epithelial involvement of the conjunctiva in 47% of cases, predominantly in the superior tarsal and fornical conjunctiva and less often in the inferior tarsal conjunctiva, caruncle, and cornea. Treatment of this condition is challenging, and map biopsy, cryotherapy, topical chemotherapy, and newer surgical methods are being used more often by our group. PMID- 15778661 TI - Orbital and periorbital microcystic adnexal carcinoma. AB - PURPOSE: To review the clinical and histopathologic features of patients with microcystic adnexal carcinoma of the orbital and periorbital tissues. METHODS: This study was designed as a noncomparative interventional case series of three patients seen and treated at two oculoplastic surgical departments and a review of the literature. Clinical presentation, histopathologic findings, outcome of surgery, and clinical recurrence are presented. RESULTS: Three patients underwent surgery for lesions temporal to the lateral canthus. All had initial histopathology diagnosed as squamous cell carcinoma but were subsequently diagnosed as microcystic adnexal carcinoma after clinical recurrence. All patients had multiple recurrences and extensive perineural spread. Two patients required radiotherapy. CONCLUSIONS: Review of the literature shows that microcystic adnexal carcinoma affecting the orbital and periorbital tissues is difficult to differentiate clinically and microscopically from other conditions including squamous cell carcinoma. Eighty-one percent of cases that have histopathology checked in the initial assessment are still misdiagnosed. A high degree of suspicion is necessary if the lesion extends beyond an apparently adequate surgical margin, multiple recurrences occur, or superficial perineural spread is present. Mohs technique is the treatment of choice and may be improved if paraffin sections or immunohistocytochemistry are performed. Wide excision should be carried out once clear margins are obtained. Radiotherapy may be necessary in cases with multiple recurrences. PMID- 15778662 TI - Malignant fibrous histiocytoma of the eyelid: differential diagnosis and management. AB - PURPOSE: Malignant fibrous histiocytoma (MFH) is a pleomorphic soft tissue sarcoma that occurs rarely in the periocular region. The purpose of this study was to present a case series of periocular MFH and to discuss the differential diagnosis and management. METHODS: This is a retrospective case review of patients diagnosed with periocular MFH from tertiary hospitals and private practices. RESULTS: Four patients, two women and two men with periocular MFH, had a mean age of 81 years (range, 72 to 85 years). All tumors were <5 cm in diameter and of storiform pleomorphic histologic subtype. One was located deep and 3 presented superficially. The initial diagnoses were sarcomatoid carcinoma, atypical fibroxanthoma, and leiomyosarcoma that were subsequently reclassified as MFH. One patient had a frozen section, 1 had fast track paraffin section margin control, and 2 had no frozen section margins at the initial excisions. Histology showed 2 negative margins, 1 close margin, and 1 positive margin that were cleared with wide local excision. Local recurrence occurred in 3 cases, and of these, 1 required orbital exenteration. Adjuvant radiotherapy was given to 3 patients. One had regional node metastases. There were no distant metastases or tumor-related deaths. Mean follow-up period was 36 +/- 35 months (median, 30; range, 1 to 84 months). CONCLUSIONS: This study highlights the difficulties in the clinicopathologic diagnosis of periocular MFH and in particular the distinction of more superficial tumors from atypical fibroxanthoma. The mainstay of treatment is complete surgical excision with wide margins, and consideration should be given to histologic margin control in addition to adjuvant radiotherapy. PMID- 15778663 TI - Horizontal eyelid movement on eyelid closure. AB - PURPOSE: When performing ptosis surgery, a suture is placed between the levator and the tarsal plate at the point of the desired peak. This placement is done while the patient's eye is closed, but its effects are observed with the patient's eye open. Unaccounted-for horizontal eyelid movement on eyelid closing may therefore adversely affect the desired outcome. The purpose of this study was to characterize the horizontal translation of the eyelid on closing the eyes. METHODS: In this experimental case series study, a randomly selected upper eyelid of normal subjects, subjects with ptosis, and subjects with exophthalmos, ages 21 through 70, was studied to determine the horizontal eyelid movement on closing the eyes by videotape analysis of the movement of dots placed above the eyelid margin and on the eyelid crease above the center of the pupil. RESULTS: The eyelid translation on closing the eye is medial and varies among subjects from 0.1 mm to 3.5 mm. The movement is usually greater at the eyelid margin than at the eyelid crease. CONCLUSIONS: The amount of horizontal eyelid translation on closing the eyes is clinically significant in many people. The clinical impact of this finding should affect how ptosis surgery is performed. The desired point of peaking of the eyelid should be marked at the beginning of the case with the eyes open and the mark used as a guideline for placing the suture that will determine the peak. PMID- 15778664 TI - Effect of darkness on upper eyelid position. AB - PURPOSE: To determine the magnitude of upper eyelid retraction induced by sudden darkness in normal subjects and in patients with Graves upper eyelid retraction before and after treatment with guanethidine drops. METHODS: The study comprised 211 control subjects (n=211 eyes) and 45 patients (n=78 eyes) with Graves upper eyelid retraction. The control subjects were divided in four age groups: 0 to 1 year, 2 to 9 years, 0 to 18 years, and 19 to 61 years. Twenty-one patients with Graves upper eyelid retraction (n=39 eyes) used guanethidine drops for 15 days. Palpebral fissure images of subjects were acquired in photopic conditions and in darkness. For both images, the distance between the mid-pupil and upper eyelid margin was measured. RESULTS: Darkness induced upper eyelid retraction in all subjects. The increment in the mid-pupil eyelid distance was greater in children. There was no significant difference between the magnitude of eyelid elevation of Graves patients and normal adults. Guanethidine drops did not abolish the eyelid reflex in Graves patients. CONCLUSIONS: Darkness provokes upper eyelid retraction in control subjects and in patients with Graves upper eyelid retraction. This effect decreases with age and does not result from sympathetic stimulation of the Muller muscle. PMID- 15778665 TI - Free tarsus autogenous graft struts for lower eyelid elevation. AB - PURPOSE: Free tarsus autogenous graft (FTG) has emerged as a useful biological spacer in lower eyelid elevation. However, it has limited use where a spacer of greater than 4 mm height is required, because at least 4 mm of upper tarsal height must be preserved to maintain upper eyelid rigidity. We describe an augmentation of FTG in its use as a spacer, by way of creating 2 parallel-placed vertical FTG struts, in contrast to interpositioning the FTG horizontally. METHODS: This case series describes the technique of harvesting and interpositioning FTG struts. Nine eyelids in 6 patients in whom FTG struts were used for lower eyelid elevation are reported. The patients ranged in age from 41 to 80 years. Underlying diagnoses included thyroid orbitopathy, previous blepharoplasty, and previous facial and eyelid thermal burns. RESULTS: FTG struts achieved an effective spacer height of up to 7 mm, and follow-up of 9 to 20 months revealed a good outcome in all 3 cases, with no complications. CONCLUSIONS: FTG struts provide an effective and lasting lower eyelid elevation when used as a spacer. A spacer graft of up to 7 mm in vertical height may be harvested from a long strip of upper eyelid tarsus that is 4 mm in vertical height. This technique would allow autogenous upper eyelid tarsus to be harvested with maximum conservation of tissue. PMID- 15778666 TI - Exposure of orbital implants wrapped with polyester-urethane after enucleation for advanced retinoblastoma. AB - PURPOSE: Enucleation is the main form of treatment for advanced retinoblastoma. The major complication of this procedure is orbital implant exposure. Different implants and wrapping materials are currently in use. The aim of the current study was to analyze the complications associated with the use of polyester urethane, an artificial dura substitute, as a wrapping material for enucleation in advanced retinoblastoma. METHODS: A retrospective review of 32 cases (28 patients), who were treated with enucleation for advanced retinoblastoma, was performed. The age of the patients ranged between 3 months and 6.7 years (median, 19 months). Additional chemotherapy was administered in 12 cases. The removed eyeball was replaced either with a silicone implant and polyester-urethane wrapping (13 cases) or hydroxyapatite, silicone-hydroxyapatite, or polyethylene implants without additional wrappings (19 cases). The follow-up period ranged from 7 months to 5.9 years (median, 22.4 months). Statistical analysis was performed using the Kaplan-Meier method. RESULTS: Single or multiple exposures occurred in 22% of cases (7/32). There were 6 exposures (46%, 6/13) in the group with polyester-urethane wrapping compared with only 1 exposure (5%, 1/19) in the implants without wrapping. This difference was statistically significant (p=0.0236). None of the other analyzed factors (additional chemotherapy, surgeon, age of the patient, or size of the implant) demonstrated a significant correlation to implant exposures. CONCLUSIONS: Wrapping of orbital implants with polyester-urethane resulted in a high rate of implant exposures after enucleation for advanced retinoblastoma. In this series, the best results were achieved with integrated implants without additional wrapping. PMID- 15778667 TI - Secondary orbital implants after evisceration: a new conjunctiva-sparing technique. AB - PURPOSE: To describe a new conjunctiva-sparing technique for secondary orbital implantation after evisceration. METHODS: Two patients with conjunctival cicatrization and a volume-deficient anophthalmic socket had implantation of an intraconal biointegratable implant. This was placed through a lateral canthal approach, after temporary disinsertion of the lateral rectus, thereby avoiding further injury to the conjunctiva. RESULTS: A good surgical outcome was achieved in both patients. There were no intraoperative or postoperative complications, and both have remained stable for nearly 2 years. CONCLUSIONS: Secondary intraconal implantation through the lateral canthal approach is safe and effective and suitable for patients in whom it is desirable to avoid a conjunctival incision. PMID- 15778668 TI - Processed fascia lata as an alternative implant material in evisceration. AB - PURPOSE: To examine the use of processed fascia lata strips as an implant material in evisceration. METHODS: This is a prospective, interventional, noncomparative case series. Eight patients underwent standard evisceration of an eye with subsequent primary insertion of processed fascia lata strips in the scleral shell. They were clinically examined at regular postoperative intervals (up to 20 months) and had computed tomography and magnetic resonance imaging scans of the orbits at the 12-month interval. The development of vascularity of the scleral shell contents was assessed with the intravenous administration of a contrast-enhancing agent. RESULTS: The fascia lata strips were incorporated without significant complications in all cases. The maintenance of adequate volume and position of the eviscerated globe was confirmed by both computed tomography and magnetic resonance imaging scans. T1-weighted magnetic resonance imaging studies with intravenous administration of contrast-enhancing agent and fat-saturation sequences revealed the development of vascularity of the scleral shell contents. CONCLUSIONS: Processed fascia lata may be a useful alternative implant material in evisceration. Possible advantages of its use may include protection against extrusion, custom volume selection and easy intraoperative manipulation. PMID- 15778669 TI - Intralesional steroid injection for management of acute idiopathic dacryoadenitis: a preliminary result. AB - PURPOSE: To evaluate the efficacy of intralesional injection of steroid as a new line of therapy in treatment of the acute form of an isolated idiopathic dacryoadenitis. METHODS: Five patients with unilateral isolated acute idiopathic dacryoadenitis were included in the study. In 2 patients, the pathology was recurrent. Intralesional injection of 2 to 4 ml betamethasone suspension (1 ml contains 2 mg betamethasone sodium phosphate and 5 mg betamethasone dipropionate) was followed by cold compression for 6 hours. Systemic nonsteroidal anti inflammatory drugs were given for 2 weeks after the injection. RESULTS: Dramatic response was achieved within 7 days after injection in all cases. No patient had recurrence during the follow-up period of 8 to 19 months. CONCLUSIONS: The excellent preliminary results make this new line of therapy highly promising. However, more study and long-term follow-up are recommended. PMID- 15778670 TI - Monocanalicular versus bicanalicular silicone intubation for nasolacrimal duct stenosis in adults. AB - PURPOSE: To compare the success rate of monocanalicular versus bicanalicular silicone intubation of incomplete nasolacrimal duct obstruction (nasolacrimal duct stenosis) in adults. METHODS: In a retrospective, nonrandomized comparative case series, 48 eyes of 44 adult patients with nasolacrimal duct stenosis underwent endoscopic probing and either bicanalicular (BCI; n=22 eyes) or monocanalicular (MCI; n=26 eyes) nasolacrimal duct intubation under general anesthesia. "Complete success" was defined as complete disappearance of the symptoms, "partial success" as improvement with some residual symptoms, and "failure" as absence of improvement or worsening of symptoms at last follow-up. The last follow-up examination included diagnostic probing and irrigation if there was not complete success. RESULTS: Patient ages ranged from 31 to 90 years (mean, 69; SD, 11.5). Forty-five tubes were removed 6 to 17 weeks (mean, 9.1; SD, 3) after surgery. Premature tube dislocation and removal occurred in one eye with BCI and in two eyes with MCI. Follow-up ranged from 6 to 52 months (mean, 14.9; SD, 8.4). The complete success rate was nearly the same in eyes with MCI (16/26, 61.53%) and BCI (13/22, 59.09%). Partial success (MCI: 8/26, 30.76%; BCI: 1/22, 4.54%) and failure (MCI: 2/26, 7.69%; BCI: 8/22, 36.36%) were, however, significantly different (p=0.010). Complications included 3 slit puncta with BCI and 4 temporary superficial punctuate keratopathy after MCI. CONCLUSIONS: MCI had virtually the same complete success rate as BCI, a higher partial success rate than BCI, and a lower failure rate than BCI in treatment of nasolacrimal duct stenosis in adults. PMID- 15778671 TI - Treatment of multiple apocrine hidrocystomas with trichloroacetic acid. AB - The apocrine hidrocystoma is a benign adenomatous cystic proliferation derived from apocrine glands, which frequently occurs in periocular tissues. These cystadenomas may occur bilaterally, in multiple disfiguring confluent groups on both the upper and lower lids. Although these lesions have been treated successfully with meticulous surgical extirpation and electrosurgery, this report describes the successful treatment of two patients, each with multiple large (>7 mm) periocular apocrine hidrocystomas by either chemical ablation of the cystic epithelium with trichloroacetic acid (TCA) or surgical excision. Examination of the cysts at 1, 3, and 6 months after TCA treatment revealed well-healed lesions without cyst recurrence. Most of the TCA-treated cysts resolved completely, without leaving any trace to clinical examination. Treatment of cysts with TCA was technically simpler and much less time-consuming than surgical excision. The treatment of large apocrine hidrocystomas with TCA is an effective and expeditious method of treating these disfiguring and recalcitrant lesions. PMID- 15778672 TI - Managing inflammatory reaction to restylane. AB - A 62-year-old woman received injections of Restylane, cross-linked hyaluronic acid, for the cosmetic filling of facial rhytids and developed a severe dermal inflammatory reaction. Hyaluronidase administered directly in the inflammatory nodules resulted in rapid resolution of pain, palpable nodularity, and visible induration possibly due to dissolution of the hyaluronic acid. PMID- 15778673 TI - Temporalis fascia in the management of gold eyelid weight extrusion. AB - A patient with a traumatic facial nerve palsy and resulting paralytic lagophthalmos underwent surgical implantation of a gold weight load, which extruded 4 weeks after surgery. Cicatricial contraction of the anterior lamella developed, and a full-thickness skin graft was used to correct the cicatrix. A second gold weight loading procedure was then undertaken with a temporalis fascia drape added to reduce the risk of extrusion. One year after surgery, there is no sign of migration or extrusion. Gold weight loading has emerged as the standard in management of paralytic lagophthalmos, with extrusion cited as the most serious complication. We propose temporalis fascia draping as an adjuvant procedure in the reimplantation of an extruded gold weight. PMID- 15778674 TI - Self-inserted intraorbital foreign bodies. AB - We report an unusual case of self-mutilation by insertion of multiple foreign bodies in the orbit. A 26-year-old female psychiatric patient with a severe emotionally unstable personality disorder was reviewed in the ophthalmology clinic. She had allegedly inserted a metal staple into her right eye 2 days previously and was complaining of pain around the right eye. Radiography showed multiple foreign bodies in the right orbit and one in the left orbit. Self mutilation in psychiatric patients has been well documented. The management of nonorganic, nontoxic intraorbital foreign bodies is discussed. A low threshold for imaging in these cases is of vital importance. PMID- 15778675 TI - Coccidioidomycosis of the eyelid. AB - A 77-year-old woman presented with an ulcerated nodule on her right lower eyelid that was unresponsive to broad-spectrum antibiotics or incision and drainage. Coccidioidomycosis was suspected because of previous serologic testing and concurrent indurated cutaneous nodules. Although skin biopsies were not diagnostic, cultures from the eyelid were positive for Coccidioides immitis. The patient was initially treated with oral fluconazole, with resolution of the eyelid and skin lesions. She ultimately completed a 6-month course of oral azole therapy, and at 18-month follow-up she was symptom free. Few cases of granulomatous coccidioidomycosis of the eyelids have been reported. The clinical features of this infection are not specific, and the diagnosis is often difficult. Presentation can mimic bacterial infection or malignant neoplasm, leading to delayed or inappropriate treatment. We describe an unusual ulcerative lesion of the eyelid that led to a diagnosis of disseminated coccidioidomycosis. PMID- 15778676 TI - Dacryocystoceles in the aftermath of Stevens-Johnson syndrome. AB - Although Stevens-Johnson syndrome has been reported to result in canalicular and nasolacrimal duct obstruction, reports of dacryocystocele formation are lacking. We describe the occurrence and management of bilateral dacryocystoceles related to Stevens-Johnson syndrome. A 45-year-old man, recovered from Stevens-Johnson syndrome, presented with bilateral medial canthal masses and intermittent overlying facial cellulitis. Examination revealed severe keratitis sicca with extensive corneal scarring, cicatricial occlusion of all puncta, and palpable medial canthal masses bilaterally. Computed tomography demonstrated bilateral cystic masses consistent with dacryocystoceles. Bilateral lacrimal sac/cyst excision was attempted, but the left cyst recurred within 6 months after surgery. A dacryocystorhinostomy was then performed, effectively marsupializing the cyst. The puncta were not canalized and stents were not placed. The masses have not recurred with 2 years of follow-up. Dacryocystoceles may arise in the setting of Stevens-Johnson syndrome. Successful management may be achieved with marsupialization to the nasal cavity (dacryocystorhinostomy). PMID- 15778677 TI - Unusual complex choristoma of the lateral canthus. AB - This report details the gross and histopathologic findings of an unusual complex choristoma at the lateral canthus. Although this lesion displays elements of hair follicle nevus, bulbar dermoid, epibulbar osseous choristoma, and accessory tragus, it does not fit neatly in any of these categories. We propose that this lesion is the result of faulty migration of pluripotential cells during embryogenesis. PMID- 15778678 TI - Conjunctival biopsy of anophthalmic socket in sarcoidosis. AB - A case of sarcoidosis was diagnosed by a random conjunctival biopsy from an anophthalmic socket. A 22-year-old black female patient presented with anterior and posterior uveitis and subconjunctival nodules in the left eye. Her right eye was enucleated due to a penetrating injury 3 years before admission. Biopsy of subconjunctival nodules in the left eye failed to demonstrate granulomatous inflammation; however, a random conjunctival biopsy from the normal-appearing anophthalmic socket conjunctiva revealed noncaseating granulomas. Conjunctival biopsy is a safe, cost-effective means of diagnosis in sarcoidosis suspects. To our knowledge, this is the first reported case of sarcoidosis confirmed with random conjunctival biopsy from an anophthalmic socket. PMID- 15778679 TI - Re: the lacrimal paradox: toward a greater understanding of success in lacrimal surgery. PMID- 15778682 TI - Re: flash visual evoked potentials in traumatic optic neuropathy. PMID- 15778690 TI - Management of atrial fibrillation. AB - Atrial fibrillation (AF) is the most common arrhythmia encountered in clinical practice. It is common in the elderly and those with structural heart disease. Clinical classification can be helpful in treatment decisions and the most widely accepted classification scheme (first episode, recurrent paroxysmal, recurrent persistent, permanent) is found in the ACC/AHA/ESC guidelines. The pathophysiology of AF remains unclear at this time. It is unlikely that a single pathophysiology is operative in all or even a majority of cases. Therapies to be considered for AF include prevention of thromboembolism, rate control, and restoration and maintenance of sinus rhythm. These therapies and specific treatments for these purposes are discussed under these headings, including a section on the relative merits of the rate control and rhythm control strategies. Risk stratification is a fundamental part of the treatment for thromboembolism. When risk warrants treatment, prevention of thromboembolism is achieved either pharmacologically with aspirin, or with warfarin or new agents like ximelagatran, or by nonpharmacological approaches. Schema to assist in risk stratification and selection of appropriate antithrombotic therapy are provided. Recent trials comparing the strategy of rate control to the strategy of rhythm control failed to demonstrate that the rhythm control approach is superior to the rate control approach in patients and therapies studied so far. Rate control is an acceptable primary line of therapy in many patients, particularly the elderly with persistent AF who are not highly symptomatic. However, the risk and benefit of each treatment modality should be individualized according to the patient circumstances and comorbidity. Algorithms to help individualize which of the two strategies to use are provided. There are a number of pharmacologic and nonpharmacologic therapies available for rhythm management of AF. Pharmacologic cardioversion is an alternative to electrical cardioversion for recent onset AF but the latter is preferred for persistent AF. Current drug therapy to maintain sinus rhythm is neither highly effective nor completely safe. An algorithm to guide selection of the most appropriate antiarrhythmic drug for an individual patient is provided. Nonpharmacologic therapies for maintenance of sinus rhythm include surgery, radiofrequency ablation, devices, and hybrid (combination) therapies. Much remains to be learned about the role and application of such therapies. Pharmacologic heart rate control can be achieved for most patients with available agents and, when it cannot, there are effective nonpharmacologic therapies. A few specific situations in which AF occurs and for which there are some special considerations are described. PMID- 15778693 TI - Alterations in transcription clusters underlie development of bladder cancer along papillary and nonpapillary pathways. AB - Bladder cancer develops in the urothelial lining from intraurothelial preneoplasia via two pathways, papillary and nonpapillary, which correspond to nonaggressive and aggressive forms of the disease. Because these two forms of cancer may develop via distinct molecular events, we examined the gene expression patterns in the development of bladder cancer from preneoplasia along papillary and nonpapillary pathways. The expression profiles of 19 pairs of RNA samples from adjacent urothelium and tumors were analyzed using cDNA microarrays. For selected genes their expressions were verified on a cohort of 251 bladder cancer patients using tissue microarray and immunohistochemistry and were related to clinicopathological parameters including follow-up data. We identified alterations in seven gene clusters controlling proliferation, differentiation, and programmed cell death that were common for papillary and nonpapillary cancer. In contrast, genes controlling cellular and stromal interactions were altered in the nonpapillary cancer. The expression levels of only two genes from this group could be used to define an aggressive form of the disease. Tumors characterized by the low expression of e-cadherin and the high expression of DNA alpha topoisomerase II had a high propensity for distant metastasis and were associated with poor survival. PMID- 15778694 TI - Mucin-producing urothelial-type adenocarcinoma of prostate: report of two cases of a rare and diagnostically challenging entity. AB - The differential diagnosis of mucin-producing adenocarcinoma of the prostate includes conventional prostatic adenocarcinoma with mucin production, secondary adenocarcinoma usually of colorectal origin and, very rarely, urothelial-type adenocarcinoma arising from either the prostatic urethra or proximal ducts. Conventional prostatic adenocarcinoma with mucin production is readily identified by routine microscopy and immunohistochemistry. The distinction between secondary adenocarcinoma and urothelial-type adenocarcinoma, however, can present a significant diagnostic challenge. In addition, documented examples of the latter in the prostate are exceptionally rare. A transurethral resection of prostate specimen and prostatic needle biopsies from two patients showing urothelial-type adenocarcinoma of the prostate were identified in our consultation files. One of the patients subsequently underwent a radical prostatectomy. Both patients had negative gastrointestinal endoscopic workups. Transurethral resection of prostate material from two patients with clinically confirmed secondary adenocarcinoma of colonic origin involving the prostate and a prostatectomy specimen with mucinous conventional prostatic adenocarcinoma were also identified for comparison purposes. Formalin-fixed, paraffin-embedded sections were stained for prostate specific antigen (PSA), prostatic acid phosphatase, carcinoembryonic antigen, cytokeratin 7, cytokeratin 20 and high molecular weight cytokeratin 34betaE12. The urothelial-type adenocarcinoma cases were diffusely positive for cytokeratin 7 and focally positive for 34betaE12 and cytokeratin 20, consistent with an origin from the urothelium of the prostatic urethra or proximal prostatic ducts. In contrast, the secondary adenocarcinoma of colonic origin cases were diffusely cytokeratin 20 positive and either negative or focally positive for cytokeratin 7 and negative for 34betaE12. The mucinous conventional prostatic adenocarcinoma was positive for PSA and prostatic acid phosphatase and negative for cytokeratin 7, cytokeratin 20 and 34betaE12. All tumors were positive for carcinoembryonic antigen. PMID- 15778698 TI - Functional implications of modifying RyR-activating peptides for membrane permeability. AB - 1. Our aim was to determine whether lipoamino acid conjugation of peptides that are high-affinity activators of ryanodine receptor (RyR) channels would (a) render the peptides membrane permeable, (b) alter their structure or (a) reduce their activity. The peptides correspond to the A region of the II-III loop of the skeletal dihydropyridine receptor. 2. The lipoamino acid conjugation increased the apparent permeability of the peptide across the Caco-2 cell monolayer by up to approximately 20-fold. 3. Nuclear magnetic resonance showed that the alpha helical structure of critical basic residues, required for optimal activation of RyRs, was retained after conjugation. 4. The conjugated peptides were more effective in enhancing resting Ca2+ release, Ca2+-induced Ca2+ release and caffeine-induced Ca2+ release from isolated sarcoplasmic reticulum (SR) than their unconjugated counterparts, and significantly enhanced caffeine-induced Ca2+ release from mechanically skinned extensor digitorum longus (EDL) fibres. 5. The effect of both conjugated and unconjugated peptides on Ca2+ release from skeletal SR was 30-fold greater than their effect on either cardiac Ca2+ release or on the Ca2+ Mg2+ ATPase. 6. A small and very low affinity effect of the peptide in slowing Ca2+ uptake by the Ca2+, Mg2+ ATPase was exacerbated by lipoamino acid conjugation in both isolated SR and in skinned EDL fibres. 7. The results show that lipoamino acid conjugation of A region peptides increases their membrane permeability without impairing their structure or efficacy in activating skeletal and cardiac RyRs. PMID- 15778699 TI - Effects of synaptotagmin reveal two distinct mechanisms of agonist-stimulated internalization of the M4 muscarinic acetylcholine receptor. AB - 1. Synaptotagmin has been reported to function in clathrin-mediated endocytosis. Here, we investigated its involvement in agonist-stimulated internalization of M4 muscarinic acetylcholine receptors exogenously expressed in human embryonic kidney (HEK-293 tsA201) cells. 2. Synaptotagmin I was present at low levels in these cells, and when overexpressed resided at the plasma membrane. 3. Synaptotagmin overexpression alone did not affect receptor internalization, but 'rescued' internalization that had been inhibited by either dominant-negative dynamin-1 or dominant-negative arrestin-2. Both normal and 'rescued' internalization were sensitive to inhibitors of clathrin-mediated endocytosis, but not to inhibitors of the function of caveolae. 4. There was no increase in AP 2 recruitment to the plasma membrane in cells overexpressing synaptotagmin. However, a mutant form of the receptor lacking a potential AP-2 recruitment motif, while being internalized normally in response to agonist stimulation, was not rescued by synaptotagmin in cells expressing dominant-negative dynamin or arrestin. 5. A mutant form of synaptotagmin (K326,327A), which binds phosphatidylinositol-4,5-bisphosphate (PIP2) much more weakly than the wild-type protein, did not rescue internalization. Furthermore, internalization was inhibited by the PH domain of phospholipase C-delta1, which sequesters PIP2, and synaptotagmin was now unable to rescue. 6. We propose that AP-2 binding to the C terminal tail of the receptor is not normally required for its endocytosis, but that the synaptotagmin-mediated rescue involves the formation of a ternary complex with the receptor and AP-2. PIP2 might play a role as an intermediary in the formation of this complex. PMID- 15778701 TI - LY294002 and rapamycin co-operate to inhibit T-cell proliferation. AB - 1. T-cell proliferation is critical for mounting an effective adaptive immune response. It is regulated by signals through the T-cell receptor, through co stimulation and through cytokines such as interleukin-2 (IL-2). Phosphatidylinositol 3-kinase (PI3K) lies downstream of each of these pathways and has been directly implicated in the regulation of lymphocyte proliferation. 2. In this study, we have shown that PI3K regulates cyclin D2 and cyclin D3, the first cell cycle proteins induced in T-cell proliferation, transcriptionally and post-transcriptionally. In T-lymphoblasts, LY294002, a PI3K inhibitor, prevents the induction of both D-type cyclin mRNA and protein, while rapamycin inhibits the induction of protein. Rapamycin inhibits mammalian target of rapamycin (mTOR), which lies downstream of PI3K. 3. Furthermore, our data show that the combination of LY294002 and rapamycin results in a co-operative inhibition of T cell proliferation. This co-operation occurs in Kit225 cells stimulated with IL 2, and also in resting peripheral blood lymphocytes stimulated with antibodies to the T-cell receptor in the presence and absence of antibodies to CD28. 4. These data indicate that PI3K regulates T-cell proliferation in response to diverse stimuli, and suggest that combinations of inhibitors, perhaps isoform-selective, may be useful as alternative immunosuppressive therapies. PMID- 15778700 TI - Excessive stimulation of poly(ADP-ribosyl)ation contributes to endothelial dysfunction in pre-eclampsia. AB - 1. Pre-eclampsia is a serious pregnancy disorder associated with widespread activation of the maternal vascular endothelium. Recent evidence implicates a role for oxidative stress in the aetiology of this condition. 2. Reactive oxygen species, particularly superoxide anions, invokes endothelial cell activation through many pathways. Oxidant-induced cell injury triggers the activation of nuclear enzyme poly(ADP-ribose) polymerase (PARP) leading to endothelial dysfunction in various pathophysiological conditions (reperfusion, shock, diabetes). 3. We have studied whether the loss of endothelial function in pre eclampsia is dependent on PARP activity. Endothelium-dependent responses of myometrial arteries were tested following exposure to either plasma from women with pre-eclampsia or normal pregnant women in the presence and absence of a novel potent inhibitor of PARP, PJ34. Additional effects of plasma and PJ34 inhibition were identified in microvascular endothelial cell cultures. 4. In myometrial arteries, PARP inhibition blocked the attenuation of endothelium dependent responses following exposure to plasma from women with pre-eclampsia. In endothelial cell cultures, plasma from pre-eclamptics induced measurable oxidative stress and a concomitant increase in PARP activity and reduction in cellular ATP. Again, these biochemical changes were reversed by PJ34. 5. These results suggest that PARP activity plays a pathogenic role in the development of endothelial dysfunction in pre-eclampsia and promotes PARP inhibition as a potential therapy in this condition. PMID- 15778702 TI - V102862 (Co 102862): a potent, broad-spectrum state-dependent blocker of mammalian voltage-gated sodium channels. AB - 1. 4-(4-Fluorophenoxy)benzaldehyde semicarbazone (V102862) was initially described as an orally active anticonvulsant with robust activity in a variety of rodent models of epilepsy. The mechanism of action was not known. We used whole cell patch-clamp techniques to study the effects of V102862 on native and recombinant mammalian voltage-gated Na+ channels. 2. V102862 blocked Na+ currents (I(Na)) in acutely dissociated cultured rat hippocampal neurons. Potency increased with membrane depolarization, suggesting a state-dependent mechanism of inhibition. There was no significant effect on the voltage dependence of activation of I(Na). 3. The dissociation constant for the inactivated state (K(I)) was approximately 0.6 microM, whereas the dissociation constant for the resting state (K(R)) was >15 microM. 4. The binding to inactivated channels was slow, requiring a few seconds to reach steady state at -80 mV. 5. The mechanism of inhibition was characterized in more detail using human embryonic kidney-293 cells stably expressing rat brain type IIA Na+ (rNa(v)1.2) channels, a major Na+ channel alpha subunit in rat hippocampal neurons. Similar to hippocampal neurons, V102862 was a potent state-dependent blocker of rNa(v)1.2 channels with a K(I) of approximately 0.4 microM and K(R) approximately 30 microM. V102862 binding to inactivated channels was relatively slow (k(+) approximately = 1.7 microM(-1) s( 1)). V102862 shifted the steady-state availability curve in the hyperpolarizing direction and significantly retarded recovery of Na+ channels from inactivation. 6. These results suggest that inhibition of voltage-gated Na+ channels is a major mechanism underlying the anticonvulsant properties of V102862. Moreover, understanding the biophysics of the interaction may prove to be useful in designing a new generation of potent Na+ channel blocker therapeutics. PMID- 15778703 TI - Blockade of HERG cardiac K+ current by antifungal drug miconazole. AB - 1. Miconazole, an imidazole antifungal agent, is associated with acquired long QT syndrome and ventricular arrhythmias. Miconazole increases the plasma concentration of QT-prolonging drugs by inhibiting the hepatic cytochrome P450 metabolic pathway, but whether it has direct effects on cardiac ion channels has not been elucidated. 2. To determine the mechanism underlying these clinical findings, we investigated the effect of miconazole on human ether-a-go-go-related gene (HERG) K+ channels. 3. HERG channels were heterologously expressed in human embryonic kidney 293 (HEK293) cells and whole-cell currents were recorded using a patch-clamp technique (23 degrees C). 4. Miconazole inhibited HERG peak tail current in a concentration-dependent manner (0.4-40 microM) with an IC50 of 2.1 microM (n=3-5 cells at each concentration, Hill coefficient 1.2). HERG block was not frequency-dependent. It required channel activation, occurred rapidly, and had very slow dissociation properties. 5. The activation curve was shifted in a negative direction (V(1/2): -9.5+/-2.3 mV in controls and -15.3+/-2.4 mV after 4 microM miconazole, P<0.05, n=6). Miconazole did not change other channel kinetics (activation, deactivation, onset of inactivation, recovery from inactivation, steady-state inactivation). 6. The S6 domain mutation, F656C, abolished the inhibitory action of miconazole on HERG current indicating that miconazole preferentially binds to an aromatic amino-acid residue within the pore-S6 region. 7. Our findings indicate that miconazole causes HERG channel block by binding to a common drug receptor, and this involves preferential binding to activated channels. Thus, miconazole prolongs the QT interval by direct inhibition of HERG channels. PMID- 15778704 TI - Synergistic and additive interactions of the cannabinoid agonist CP55,940 with mu opioid receptor and alpha2-adrenoceptor agonists in acute pain models in mice. AB - 1. Cannabinoid receptor agonists elicit analgesic effects in acute and chronic pain states via spinal and supraspinal pathways. We investigated whether the combination of a cannabinoid agonist with other classes of antinociceptive drugs exerted supra-additive (synergistic) or additive effects in acute pain models in mice. 2. The interactions between the cannabinoid agonist CP55,940, alpha2 adrenoceptor agonist dexmedetomidine and mu-opioid receptor agonist morphine were evaluated by isobolographic analysis of antinociception in hot plate (55 degrees C) and tail flick assays in conscious male Swiss mice. Drug interactions were examined by administering fixed-ratio combinations of agonists (s.c.) in 1:1, 3:1 and 1:3 ratios of their respective ED50 fractions. 3. CP55,940, dexmedetomidine and morphine all caused dose-dependent antinociception. In the hot plate and tail flick assays, ED50 values (mg kg(-1)) were CP55,940 1.13 and 0.51, dexmedetomidine 0.066 and 0.023, and morphine 29.4 and 11.3, respectively. Synergistic interactions existed between CP55,940 and dexmedetomidine in the hot plate assay, and CP55,940 and morphine in both assays. Additive interactions were found for CP55,940 and dexmedetomidine in the tail flick assay, and dexmedetomidine and morphine in both assays. 4. Thus, an alpha2-adrenoceptor agonist or mu opioid receptor agonist when combined with a cannabinoid receptor agonist showed significant synergy in antinociception in the hot plate test. However, for the tail flick nociceptive response to heat, only cannabinoid and mu opioid receptor antinociceptive synergy was demonstrated. If these results translate to humans, then prudent selection of dose and receptor-specific agonists may allow an improved therapeutic separation from unwanted side effects. PMID- 15778705 TI - Sequence-dependent stimulation of the mammalian innate immune response by synthetic siRNA. AB - Short interfering RNAs (siRNAs) that mediate specific gene silencing through RNA interference (RNAi) are widely used to study gene function and are also being developed for therapeutic applications. Many nucleic acids, including double- (dsRNA) and single-stranded RNA (ssRNA), can stimulate innate cytokine responses in mammals. Despite this, few studies have questioned whether siRNA may have a similar effect on the immune system. This could significantly influence the in vivo application of siRNA owing to off-target effects and toxicities associated with immune stimulation. Here we report that synthetic siRNAs formulated in nonviral delivery vehicles can be potent inducers of interferons and inflammatory cytokines both in vivo in mice and in vitro in human blood. The immunostimulatory activity of formulated siRNAs and the associated toxicities are dependent on the nucleotide sequence. We have identified putative immunostimulatory motifs that have allowed the design of siRNAs that can mediate RNAi but induce minimal immune activation. PMID- 15778706 TI - Wnt signalling induces maturation of Paneth cells in intestinal crypts. AB - Wnt signalling, which is transduced through beta-catenin/TCF4, maintains the undifferentiated state of intestinal crypt progenitor cells. Mutational activation of the pathway initiates the adenomacarcinoma sequence. Whereas all other differentiated epithelial cells migrate from the crypt onto the villus, Paneth cells home towards the source of Wnt signals--that is, the crypt bottom. Here, we show that expression of a Paneth gene programme is critically dependent on TCF4 in embryonic intestine. Moreover, conditional deletion of the Wnt receptor Frizzled-5 abrogates expression of these genes in Paneth cells in the adult intestine. Conversely, adenomas in Apc-mutant mice and colorectal cancers in humans inappropriately express these Paneth-cell genes. These observations imply that Wnt signals in the crypt can separately drive a stem-cell/progenitor gene programme and a Paneth-cell maturation programme. In intestinal cancer, both gene programmes are activated simultaneously. PMID- 15778707 TI - An empirical test of the mutational landscape model of adaptation using a single stranded DNA virus. AB - The primary impediment to formulating a general theory for adaptive evolution has been the unknown distribution of fitness effects for new beneficial mutations. By applying extreme value theory, Gillespie circumvented this issue in his mutational landscape model for the adaptation of DNA sequences, and Orr recently extended Gillespie's model, generating testable predictions regarding the course of adaptive evolution. Here we provide the first empirical examination of this model, using a single-stranded DNA bacteriophage related to phiX174, and find that our data are consistent with Orr's predictions, provided that the model is adjusted to incorporate mutation bias. Orr's work suggests that there may be generalities in adaptive molecular evolution that transcend the biological details of a system, but we show that for the model to be useful as a predictive or inferential tool, some adjustments for the biology of the system will be necessary. PMID- 15778708 TI - Genetic dissection and prognostic modeling of overt stroke in sickle cell anemia. AB - Sickle cell anemia (SCA) is a paradigmatic single gene disorder caused by homozygosity with respect to a unique mutation at the beta-globin locus. SCA is phenotypically complex, with different clinical courses ranging from early childhood mortality to a virtually unrecognized condition. Overt stroke is a severe complication affecting 6-8% of individuals with SCA. Modifier genes might interact to determine the susceptibility to stroke, but such genes have not yet been identified. Using Bayesian networks, we analyzed 108 SNPs in 39 candidate genes in 1,398 individuals with SCA. We found that 31 SNPs in 12 genes interact with fetal hemoglobin to modulate the risk of stroke. This network of interactions includes three genes in the TGF-beta pathway and SELP, which is associated with stroke in the general population. We validated this model in a different population by predicting the occurrence of stroke in 114 individuals with 98.2% accuracy. PMID- 15778709 TI - Reverse engineering of regulatory networks in human B cells. AB - Cellular phenotypes are determined by the differential activity of networks linking coregulated genes. Available methods for the reverse engineering of such networks from genome-wide expression profiles have been successful only in the analysis of lower eukaryotes with simple genomes. Using a new method called ARACNe (algorithm for the reconstruction of accurate cellular networks), we report the reconstruction of regulatory networks from expression profiles of human B cells. The results are suggestive a hierarchical, scale-free network, where a few highly interconnected genes (hubs) account for most of the interactions. Validation of the network against available data led to the identification of MYC as a major hub, which controls a network comprising known target genes as well as new ones, which were biochemically validated. The newly identified MYC targets include some major hubs. This approach can be generally useful for the analysis of normal and pathologic networks in mammalian cells. PMID- 15778710 TI - AlphaCaMKII autophosphorylation contributes to rapid learning but is not necessary for memory. AB - Autophosphorylation of alpha calcium-calmodulin-dependent kinase II (alphaCaMKII) has been proposed to be the key event in memory storage. We tested this hypothesis with autophosphorylation-deficient mutant mice in hippocampus- and amygdala-dependent learning and memory tasks and found that the autophosphorylation of alphaCaMKII was required for rapid learning but was not essential for memory. We conclude that alphaCaMKII autophosphorylation contributes to single-trial learning but is dispensable for memory. PMID- 15778711 TI - Virtual lesions of the anterior intraparietal area disrupt goal-dependent on-line adjustments of grasp. AB - Adaptive motor behavior requires efficient error detection and correction. The posterior parietal cortex is critical for on-line control of reach-to-grasp movements. Here we show a causal relationship between disruption of cortical activity within the anterior intraparietal sulcus (aIPS) by transcranial magnetic stimulation (TMS) and disruption of goal-directed prehensile actions (either grip size or forearm rotation, depending on the task goal, with reaching preserved in either case). Deficits were elicited by applying TMS within 65 ms after object perturbation, which attributes a rapid control process on the basis of visual feedback to aIPS. No aperture deficits were produced when TMS was applied to a more caudal region within the intraparietal sulcus, to the parieto-occipital complex (putative V6, V6A) or to the hand area of primary motor cortex. We contend that aIPS is critical for dynamic error detection during goal-dependent reach-to-grasp action that is visually guided. PMID- 15778712 TI - Protocadherin Celsr3 is crucial in axonal tract development. AB - In the embryonic CNS, the development of axonal tracts is required for the formation of connections and is regulated by multiple genetic and microenvironmental factors. Here we show that mice with inactivation of Celsr3, an ortholog of Drosophila melanogaster flamingo (fmi; also known as starry night, stan) that encodes a seven-pass protocadherin, have marked, selective anomalies of several major axonal fascicles, implicating protocadherins in axonal development in the mammalian CNS for the first time. In flies, fmi controls planar cell polarity (PCP) in a frizzled-dependent but wingless-independent manner. The neural phenotype in Celsr3 mutant mice is similar to that caused by inactivation of Fzd3, a member of the frizzled family. Celsr3 and Fzd3 are expressed together during brain development and may act in synergy. Thus, a genetic pathway analogous to the one that controls PCP is key in the development of the axonal blueprint. PMID- 15778713 TI - G protein betagamma directly regulates SNARE protein fusion machinery for secretory granule exocytosis. AB - The activation of G protein-coupled receptors (GPCRs) can result in an inhibition of Ca(2+)-dependent hormone and neurotransmitter secretion. This has been attributed in part to G protein inhibition of Ca(2+) influx. However, a frequently dominant inhibitory effect, of unknown mechanism, also occurs distal to Ca(2+) entry. Here we characterize direct inhibitory actions of G protein betagamma (Gbetagamma) on Ca(2+)-triggered vesicle exocytosis in permeable PC12 cells. Gbetagamma inhibition was rapid (<1 s) and was attenuated by cleavage of synaptosome-associated protein of 25 kD (SNAP25). Gbetagamma bound soluble N ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes, and binding was reduced to SNARE complexes containing cleaved SNAP25 or by Ca(2+) dependent synaptotagmin binding. Here we show inhibitory coupling between GPCRs and vesicle exocytosis mediated directly by Gbetagamma interactions with the Ca(2+)-dependent fusion machinery. PMID- 15778715 TI - Duality of fatigue failures of materials caused by Poisson defect statistics of competing failure modes. AB - The statistical nature of failures of components subjected to fatigue loading is a major challenge in engineering applications. Here, we report an interesting fatigue phenomenon in which there is a 'duality' of the S-N fatigue data (S is stress and N is cycles to failure)--the grouping of data into two distinct S-N curves as a result of two competing failure modes. The proportions of failure by either mode are about the same and one cannot predict which failure mode will eventually dominate. This means that fatigue lives may be unpredictable in materials under certain application conditions and the consequences can be disastrous. Based on Poisson defect statistics and Monte Carlo simulations, we show that this phenomenon can arise from the Poisson distribution of low populations of microscale defects. An unexpected finding is that there should be a combination of average defect density and specimen area at which this phenomenon is observable in materials having sparsely populated defects. PMID- 15778714 TI - Spin-conserving carrier recombination in conjugated polymers. AB - The ultimate efficiency of polymer light-emitting diodes is limited by the fraction of charges recombining in the molecular singlet manifold. We address the question of whether this fraction can principally exceed the fundamental limit set down by spin statistics, which requires the possibility of spin changes during exciton formation. Sensitized phosphorescence at 4-300 K enables a direct quantification of spin conversion in coulombically bound electron-hole pairs, the precursors to exciton formation. These are stabilized in external electric fields over times relevant to carrier transport, capture and recombination in devices. No interconversion of exciton intermediates between singlet and triplet configurations is observed. Static magnetic fields are equally unable to induce spin mixing in electroluminescence. Our observations imply substantial exchange splitting at all times during carrier capture. Prior statements regarding increased singlet yields above 25% merely on the basis of higher singlet than triplet formation rates should therefore be re-examined. PMID- 15778716 TI - Trans-interface diffusion-controlled coarsening. AB - Accurate theoretical predictions of the volume-fraction dependence during diffusion-controlled coarsening of a polydisperse assembly of particles have proved difficult. Here, a new model of coarsening is presented, involving diffusive transport through the coherent interface between ordered and disordered phases, which atomistic calculations show has a ragged structure. The interface is a diffusion bottleneck when the ordered phase is dispersed. It is predicted that the square of the average radius grows linearly with time, that the depletion of solute decreases as the inverse square-root of time, and that there is no effect of volume fraction on kinetics and the scaled particle-size distributions. These differ dramatically from predictions of modern theories of diffusion-controlled coarsening. Data on coarsening in Ni-Al alloys is examined. We show that no other theory is consistent with the experimentally observed absence of an effect of volume fraction on coarsening of ordered gamma' (Ni3Al) precipitates in a disordered Ni-Al (gamma) matrix, and the strong volume-fraction dependence of coarsening of gamma precipitates in an ordered gamma' matrix. PMID- 15778717 TI - Quantum criticality and universal scaling of a quantum antiferromagnet. AB - Quantum effects dominate the behaviour of many diverse materials. Of particular current interest are those systems in the vicinity of a quantum critical point (QCP). Their physical properties are predicted to reflect those of the nearby QCP with universal features independent of the microscopic details. The prototypical QCP is the Luttinger liquid (LL), which is of relevance to many quasi-one dimensional materials. The magnetic material KCuF3 realizes an array of weakly coupled spin chains (or LLs) and thus lies close to but not exactly at the LL quantum critical point. By using inelastic neutron scattering we have collected a complete data set of the magnetic correlations of KCuF3 as a function of momentum, energy and temperature. The LL description is found to be valid over an extensive range of these parameters, and departures from this behaviour at high and low energies and temperatures are identified and explained. PMID- 15778718 TI - Mechanism of nonhomologous end-joining in mycobacteria: a low-fidelity repair system driven by Ku, ligase D and ligase C. AB - DNA double-strand breaks (DSBs) can be repaired either via homologous recombination (HR) or nonhomologous end-joining (NHEJ). Both pathways are operative in eukaryotes, but bacteria had been thought to rely on HR alone. Here we provide direct evidence that mycobacteria have a robust NHEJ pathway that requires Ku and a specialized polyfunctional ATP-dependent DNA ligase (LigD). NHEJ of blunt-end and complementary 5'-overhang DSBs is highly mutagenic ( approximately 50% error rate). Analysis of the recombination junctions ensuing from individual NHEJ events highlighted the participation of several DNA end remodeling activities, including template-dependent fill-in of 5' overhangs, nontemplated addition of single nucleotides at blunt ends, and nucleolytic resection. LigD itself has the template-dependent and template-independent polymerase functions in vitro that compose the molecular signatures of NHEJ in vivo. Another ATP-dependent DNA ligase (LigC) provides a backup mechanism for LigD-independent error-prone repair of blunt-end DSBs. We speculate that NHEJ allows mycobacteria to evade genotoxic host defense. PMID- 15778719 TI - The HEAT repeat protein Blm10 regulates the yeast proteasome by capping the core particle. AB - Proteasome activity is fine-tuned by associating the proteolytic core particle (CP) with stimulatory and inhibitory complexes. Although several mammalian regulatory complexes are known, knowledge of yeast proteasome regulators is limited to the 19-subunit regulatory particle (RP), which confers ubiquitin dependence on proteasomes. Here we describe an alternative proteasome activator from Saccharomyces cerevisiae, Blm10. Synthetic interactions between blm10Delta and other mutations that impair proteasome function show that Blm10 functions together with proteasomes in vivo. This large, internally repetitive protein is found predominantly within hybrid Blm10-CP-RP complexes, representing a distinct pool of mature proteasomes. EM studies show that Blm10 has a highly elongated, curved structure. The near-circular profile of Blm10 adapts it to the end of the CP cylinder, where it is properly positioned to activate the CP by opening the axial channel into its proteolytic chamber. PMID- 15778720 TI - Angiopoietin-related growth factor antagonizes obesity and insulin resistance. AB - Angiopoietin-related growth factor (AGF), a member of the angiopoietin-like protein (Angptl) family, is secreted predominantly from the liver into the systemic circulation. Here, we show that most (>80%) of the AGF-deficient mice die at about embryonic day 13, whereas the surviving AGF-deficient mice develop marked obesity, lipid accumulation in skeletal muscle and liver, and insulin resistance accompanied by reduced energy expenditure relative to controls. In parallel, mice with targeted activation of AGF show leanness and increased insulin sensitivity resulting from increased energy expenditure. They are also protected from high-fat diet-induced obesity, insulin resistance and nonadipose tissue steatosis. Hepatic overexpression of AGF by adenoviral transduction, which leads to an approximately 2.5-fold increase in serum AGF concentrations, results in a significant (P < 0.01) body weight loss and increases insulin sensitivity in mice fed a high-fat diet. This study establishes AGF as a new hepatocyte-derived circulating factor that counteracts obesity and related insulin resistance. PMID- 15778721 TI - A new transgene reporter for in vivo magnetic resonance imaging. AB - We report a new platform technology for visualizing transgene expression in living subjects using magnetic resonance imaging (MRI). Using a vector, we introduced an MRI reporter, a metalloprotein from the ferritin family, into specific host tissues. The reporter is made superparamagnetic as the cell sequesters endogenous iron from the organism. In this new approach, the cells construct the MRI contrast agent in situ using genetic instructions introduced by the vector. No exogenous metal-complexed contrast agent is required, thereby simplifying intracellular delivery. We used a replication-defective adenovirus vector to deliver the ferritin transgenes. Following focal inoculation of the vector into the mouse brain, we monitored the reporter activity using in vivo time-lapse MRI. We observed robust contrast in virus-transduced neurons and glia for several weeks. This technology is adaptable to monitor transgene expression in vivo in many tissue types and has numerous biomedical applications, such as visualizing preclinical therapeutic gene delivery. PMID- 15778722 TI - Somatostatin regulates brain amyloid beta peptide Abeta42 through modulation of proteolytic degradation. AB - Expression of somatostatin in the brain declines during aging in various mammals including apes and humans. A prominent decrease in this neuropeptide also represents a pathological characteristic of Alzheimer disease. Using in vitro and in vivo paradigms, we show that somatostatin regulates the metabolism of amyloid beta peptide (Abeta), the primary pathogenic agent of Alzheimer disease, in the brain through modulating proteolytic degradation catalyzed by neprilysin. Among various effector candidates, only somatostatin upregulated neprilysin activity in primary cortical neurons. A genetic deficiency of somatostatin altered hippocampal neprilysin activity and localization, and increased the quantity of a hydrophobic 42-mer form of Abeta, Abeta(42), in a manner similar to presenilin gene mutations that cause familial Alzheimer disease. These results indicate that the aging-induced downregulation of somatostatin expression may be a trigger for Abeta accumulation leading to late-onset sporadic Alzheimer disease, and suggest that somatostatin receptors may be pharmacological-target candidates for prevention and treatment of Alzheimer disease. PMID- 15778723 TI - Immune restoration following hematopoietic stem cell transplantation: an evolving target. AB - Hematopoietic stem cell transplantation (HSCT) is the definitive cure for many malignant and nonmalignant diseases. However, delays in immune reconstitution (IR) following HSCT significantly limit the success of transplantation and increase the risk for infection and disease relapse in the transplant recipient. Therefore, ways to measure and to manipulate immune recovery following HSCT are emerging and their success depends directly upon an enhanced understanding for the underlying mechanisms responsible for reconstituted immunity and hematopoiesis. Recent discoveries in the activation, function, and regulation of dendritic cell (DC), natural killer (NK) cell, and T-lymphocyte subtypes have been critical in developing immunotherapies used to prevent graft-versus-host disease and to enhance graft-versus-leukemia. For example, regulatory T cells that induce tolerance and NK receptor-tumor ligand disparities that result in tumor lysis are being used to minimize GVHD and tumor burden, respectively. Furthermore, expansion and modulation of immune effector cells are being used to augment hematopoietic and immune recovery and to decrease transplant-related toxicity in the transplant recipient. Specifically, DC expansion and incorporation into antitumor and anti-microbial vaccines is fast approaching application into clinical trials. This paper will review our current understanding for IR following HSCT and the novel ways in which to restore immune function and decrease transplant-related toxicity in the transplant recipient. PMID- 15778724 TI - A new case of IPEX receiving bone marrow transplantation. PMID- 15778725 TI - Prospective randomised trial of amifostine cytoprotection in myeloma patients undergoing high-dose melphalan conditioned autologous stem cell transplantation. AB - In this prospective multicentre trial, 90 patients undergoing autologous stem cell transplantation (ASCT) were randomised to receive (n=43) or not receive (n=47) amifostine 910 mg/m(2) prior to melphalan 200 mg/m(2). Patients were monitored for regimen-related toxicity, engraftment, supportive care, response and survival. Both groups underwent ASCT at a median of 8 months from diagnosis and were matched for disease characteristics, prior therapy and pre-ASCT disease responsiveness. Amifostine infusional side-effects were frequent, occurring in 65% of patients, but of mild severity. Amifostine use was associated with a reduction in the median grade of oral mucositis (1 vs 2, P=0.01) and the frequency of severe (WHO grades 3 or 4) mucositis (12 vs 33%, P=0.02), but no reduction in the requirement for parenteral nutrition or analgesic use. Conversion to complete remission post-ASCT occurred in 30 and 14% of the amifostine and control groups, respectively (P=0.09). With a median follow-up of 35 months, there was no statistically significant difference between the median progression-free or overall survival times for the two groups. We conclude that amifostine can be safely administered prior to high-dose melphalan and significantly reduces the frequency and severity of therapy-induced oral mucositis. PMID- 15778726 TI - Donor Y chromosome in renal carcinoma cells of a female BMT recipient: visualization of putative BMT-tumor hybrids by FISH. PMID- 15778727 TI - Long-term outcome of allogeneic or autologous haemopoietic cell transplantation for acute lymphoblastic leukaemia in second remission in children. GETMON experience 1983-1998. AB - We present a retrospective study of long-term outcome and predictive factors of survival and relapse in 219 paediatric patients with acute lymphoblastic leukaemia (ALL) in second remission. They received allogeneic (allo) or autologous (auto) haemopoietic cell transplantation (HCT) depending on the availability of a matched sibling donor. The probability of event-free survival (EFS) for the total patient group was 0.35+0.03 at 14 years. No significant differences were observed for EFS between allo- and auto-HCT: 0.39+0.05 vs 0.32+0.04 (P=0.43). A better EFS was seen in patients with a late relapse (LR) (P=0.06 and 0.02, for allogeneic and autologous respectively). Significantly better EFS was observed in allo-HCT patients under 10 years of age and in auto HCT patients with leukocytes at diagnosis below 25 x 109/l and late relapse. Predictive factors of failure in both groups were early relapse (ER), medullary relapse and age over 10 years. The probability of relapse (RP) for the total group of patients was 0.57+0.03, and it was significantly higher in auto-HCT patients: 0.65+0.04 vs 0.42+0.06 (P=0.002). Factors predictive for relapse were medullary and early relapse, auto-HCT and WBC >25 x 109/l at diagnosis. PMID- 15778728 TI - Oligotide, a defibrotide derivative, protects human microvascular endothelial cells against fludarabine-induced activation, damage and allogenicity. AB - Fludarabine is a nonmyeloablative immunosuppressant increasingly used as a component of alternative reduced-intensity conditioning regimens prior to allogeneic stem cell transplantation (SCT). However, we have previously shown that 2-fluoroadenine 9-beta-D-arabinofuranoside (F-Ara) as the active metabolized form of fludarabine induces damage, activation and allogenicity in human microvascular endothelial cells (HMEC). We had also identified the pharmaceutic compound Defibrotide (DF), originally used in the treatment of veno-occlusive disease and thrombotic microangiopathy, as being protective against F-Ara-induced dysfunction of HMEC, importantly, without affecting the antileukemic effect of F Ara. In the present report, we show that a recently developed derivative of DF, Oligotide, similarly downregulates F-Ara-induced activation and damage of HMEC as well as their antigenicity for allogeneic CD8+ T cells. In addition, Oligotide could also block F-Ara-mediated transendothelial migration of peripheral blood cells across the HMEC barrier. Taken together, these observations argue for a potential clinical use of both DF and Oligotide in pre transplant conditioning. PMID- 15778729 TI - Association of post-thaw viable CD34+ cells and CFU-GM with time to hematopoietic engraftment. AB - In all, 78 peripheral hematopoietic progenitor cell collections from 52 patients were evaluated using our previously published validated post-thaw assays at the time of collection and following transplantation by assessment of viable CD34(+) cells, and granulocyte-macrophage colony-forming units (CFU-GM) cryopreserved in quality control vials. The median (range) post-thaw recovery of viable CD34(+) cells and CFU-GM was 66.4% (36.1-93.6%) and 63.0% (28.6-85.7%), respectively, which did not show significant correlation with the engraftment of either neutrophils (P=0.136 and 0.417, respectively) or platelets (P=0.88 and 0.126, respectively). However, the reinfused viable CD34(+) cells/kg of patient weight pre- or post-cryopreservation showed significant correlation to engraftment of neutrophils (P=0.0001 and 0.001, respectively) and platelets (P=0.023 and 0.010, respectively), whereas CFU-GM pre- or post-cryopreservation was significantly correlated to neutrophils (P=0.011 and 0.007, respectively) but not to platelets (P=0.112 and 0.100, respectively). The results show that post-cryopreservation assessment of viable CD34(+) cells or CFU-GM is as reliable a predictor of rapid engraftment as that of pre-cryopreservation measures. Therefore, the post cryopreservation number of viable CD34(+) cells or CFU-GM should be used to eliminate the risks of unforeseen cell loss that could occur during cryopreservation or long-term storage. PMID- 15778730 TI - Haplotype mismatched transplantation using high doses of peripheral blood CD34+ cells together with stratified conditioning regimens for high-risk adult acute myeloid leukemia patients: a pilot study in a single Korean institution. AB - A total of 11 high-risk Korean acute myeloid leukemia (AML) patients received stem cell transplantation from human leukocyte antigen (HLA) haploidentical donors. Specifically, for eight patients with 2-3 mismatched antigens and bidirectional vectors, we used a newly designed conditioning regimen that consists of total body irradiation, busulfex, ATG, and fludarabine. The median number of CD34+ cells infused was 15.4 x 10(6)/kg (range, 8-21.2). These patients received neither graft-versus-host disease (GvHD) prophylaxis nor post transplantation G-CSF. All of the patients who were followed up for a median of 6 months (range, 17 days-28 months) showed stable primary engraftment and had no acute GvHD or transplant-related mortality for 100 days post transplant. Three patients with high-risk or refractory disease eventually died in relapse, even with GvH-directed NK alloreactivity. However, the patients in complete remission (CR), with the exception of one patient who is alive at 18 months EFS, died at 4, 6, and 8 months post transplantation due to infections that were associated with delayed immune recovery. Our findings suggest that haploidentical transplantation represents a feasible treatment for patients with high-risk AML in CR, provided that a plan for the enhancement of immune recovery is implemented. PMID- 15778731 TI - Allogeneic transplantation for hepatosplenic alphabeta T-cell lymphoma. PMID- 15778732 TI - Successful allogeneic bone marrow transplantation in a patient with active Coccidioidomycosis. PMID- 15778733 TI - Progesterone enhances cytokine-stimulated nitric oxide synthase II expression and cell death in human breast cancer cells. AB - The presence of hormone receptors is related to survival outcome in breast cancer. Previous results from our laboratory established a correlation between the presence of nitric oxide synthase II (NOSII) and nitric oxide (NO) production with progesterone receptors in a series of human breast tumours. Furthermore, this was directly related to a lower tumour grade and a lower proliferation rate of the tumour cells. To examine these results in further detail, the effect of progesterone (Pg) and 17beta-oestradiol (E2) on NOSII expression was analysed in the human breast cancer cell line MCF-7. By Northern blot and promoter activity, we show that a cytokine mix (TNF-alpha, IL-beta, and IFN-gamma) induces NOSII transcription after 6 h stimulation. In the absence of cytokines, neither hormone affects NOSII expression. However, Pg but not E2, enhances cytokine-induced NOSII transcription as well as NO synthesis, mainly by cooperation with gamma interferon. The increase in NO accumulation in the media induced by addition of Pg to the cytokine treatment significantly increases cell death, mainly accounted for by apoptosis, as compared to the effect of cytokines alone. Our findings help clarify the role of steroid hormones in NOSII expression as well as the effect on cell viability and may suggest novel approaches towards hormonotherapy and the treatment of cancer. PMID- 15778734 TI - Decreasing the infusion rate reduces the proarrhythmic risk of NS-7: confirming the relevance of short-term variability of repolarisation in predicting drug induced torsades de pointes. AB - 1 The rate of infusion has been suggested to be important for drug-induced torsades de pointes (TdP) arrhythmias. We investigated the repolarisation prolonging effects and proarrhythmic properties of NS-7, a neuroprotective drug in development, using two different infusion rates. 2 A fast (5 min intravenously (i.v.)) escalating dosing regimen (0.3 and 3.0 mg kg(-1), n=4) of NS-7 was investigated in anaesthetised control dogs in sinus rhythm (SR). This was compared to a slow infusion (60 min i.v.) of one dose (3.0 mg kg(-1), n=4) NS-7. The similar dosing regimens were investigated in anaesthetised dogs with chronic, complete AV block (CAVB), an animal model of TdP (n=6). 3 No electrophysiological effects were seen after 0.3 mg kg(-1) NS-7. Fast infusion of 3.0 mg kg(-1) caused prolongation of repolarisation, for example, heart rate corrected QT interval (QT(c)): in SR: 6+/-1%; in CAVB: 10+/-7%, which was accompanied by TdP in three of six CAVB dogs. No TdP were seen in SR dogs. 4 Slow infusion did not cause TdP in the same CAVB dogs, although NS-7 caused repolarisation to prolong with a similar magnitude (QT(c): 12+/-7%) as in the fast-infusion experiment. 5 Short term variability (STV) is a novel parameter for the prediction of drug-induced TdP analysing the beat-to-beat variability of repolarisation. STV was only increased after the fast infusion in CAVB dogs (2.6+/-0.3 versus 6.0+/-1.4 ms, P<0.05), while there was no increase (2.1+/-0.2 versus 2.5+/-1.0 ms) after the slow infusion of NS-7. 6 Peak plasma concentrations attained were lower in slow (0.5+/-0.1 microg ml(-1) after 50 min) than in fast-infusion regimen (2.1+/-0.4 microg ml(-1) after 5 min; P<0.05). 7 The results support the conclusion that limiting peak plasma concentration by decreasing the rate of infusion of NS-7 reduces the proarrhythmic risk despite comparable prolongation in repolarisation parameters. The relevance of STV in predicting drug-induced TdP was confirmed. PMID- 15778735 TI - Megakaryocyte apoptosis: sorting out the signals. PMID- 15778736 TI - Emerging roles for cyclooxygenase-2 in gastrointestinal mucosal defense. AB - The development of selective inhibitors of cyclooxygenase-2 (COX-2) was based on the concept that this enzyme played little, if any, role in modulating the ability of the gastrointestinal (GI) tract to resist and respond to injury. There is now overwhelming evidence that this is far from true. Indeed, COX-2 mediates several of the most important components of 'mucosal defense', contributes significantly to the resolution of GI inflammation and plays a crucial role in regulating ulcer healing. COX-2 also contributes to long-term changes in GI function after bouts of inflammation. PMID- 15778737 TI - Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis. AB - 1 We have previously demonstrated that nitric oxide (NO) triggers CD34(+)-derived megakaryocyte apoptosis. We here show that prostacyclin (PGI(2)) inhibits PAPA/NO induced megakaryocyte death detected by fluorescent microscopy and flow cytometry. 2 The cAMP-specific phosphodiesterase inhibitor, Ro 20-1724, and the permeable analog dibutyryl-cAMP also delayed apoptosis. PGI(2) effect was fully prevented when adenylyl cyclase activity was suppressed by SQ 22536, and partially reversed by the permeable protein kinase A inhibitor PKI 14-22 amide. ELISA showed that while both PGI(2) and NO alone or synergistically raised cAMP, only NO was able to increase intracellular cGMP levels. 3 Treatment of megakaryocytes with PGI(2) abolished both basal and NO-raised cGMP levels. Addition of 8-pCPT-cGMP or activation of soluble guanylyl cyclase by BAY 41-2272 induced cell death in a concentration-dependent manner, and ODQ, an inhibitor of guanylyl cyclase, prevented both PAPA/NO- or BAY 41-2272-induced apoptosis. Specific cGMP phosphodiesterase inhibition by Zaprinast or suppression of adenylyl cyclase by SQ 22536 enhanced the PAPA/NO proapoptotic effect. 4 PGI(2) completely inhibited NO-mediated generation and the increased activity of the cleaved form of caspase-3. 5 In conclusion, our results demonstrate that contrary to their well-known direct and synergistic inhibitory effects on platelets, PGI(2) and NO regulate opposite megakaryocyte survival responses through a delicate balance between intracellular cyclic nucleotide levels and caspase-3 activity control. PMID- 15778738 TI - Expression of functional NK1 receptors in human alveolar macrophages: superoxide anion production, cytokine release and involvement of NF-kappaB pathway. AB - 1 Substance P (SP) is deeply involved in lung pathophysiology and plays a key role in the modulation of inflammatory-immune processes. We previously demonstrated that SP activates guinea-pig alveolar macrophages (AMs) and human monocytes, but a careful examination of its effects on human AMs is still scarce. 2 This study was undertaken to establish the role of SP in human AM isolated from healthy smokers and non-smokers, by evaluating the presence of tachykinin NK(1) receptors (NK-1R) and SP's ability to induce superoxide anion (O(2)(-)) production and cytokine release, as well as activation of the nuclear factor kappaB (NF-kappaB) pathway. 3 By Western blot analysis and immunofluorescence, we demonstrate that authentic NK-1R are present on human AMs, a three-fold enhanced expression being observed in healthy smokers. These NK-1R are functional, as SP and NK(1) agonists dose-dependently induce O(2)(-) production and cytokine release. In AMs from healthy smokers, SP evokes an enhanced respiratory burst and a significantly increased release of tumor necrosis factor-alpha as compared to healthy non-smokers, but has inconsistent effects on IL-10 release. The NK(1) selective antagonist CP 96,345 ((2S,3S)-cis-2-diphenylmethyl-N[(2-methoxyphenyl) methyl]-1-azabicyclo-octan-3-amine)) competitively antagonized SP-induced effects. 4 SP activates the transcription factor NF-kappaB, a three-fold increased nuclear translocation being observed in AMs from healthy smokers. This effect is receptor-mediated, as it is reproduced by the NK(1) selective agonist [Sar(9)Met(O(2))(11)]SP and reverted by CP 96,345. 5 These results clearly indicate that human AMs possess functional NK-1R on their surface, which are upregulated in healthy smokers, providing new insights on the mechanisms involved in tobacco smoke toxicity. PMID- 15778739 TI - Antagonism of ATP responses at P2X receptor subtypes by the pH indicator dye, Phenol red. AB - 1 Many types of culture media contain a pH-sensitive dye. One commonly occurring dye, Phenol red sodium (Na(+)) salt, was tested for blocking activity at rat P2X(1-4) receptors (P2X(1-4)Rs) expressed in Xenopus oocytes. 2 Phenol red Na(+) salt antagonised adenosine 5'-triphosphate (ATP) responses at P2X(1)R (IC(50), 3 microM) and, at higher concentrations, also blocked P2X(2)R and P2X(3)R. Phenol red Na(+)-salt, purified of lipophilic contaminants, blocked P2X(1)R and P2X(3)R by acting as an insurmountable antagonist. 3 Two lipophilic extracts of Phenol red antagonised ATP responses at P2XRs. Extract A was a potent antagonist at P2X(1)R (IC(50), 1.4 microM), whereas extract B was a potent antagonist at P2X(3)R (IC(50), 4.1 microM). A bisphenolic compound (RS151030) found in these extracts was a potent antagonist at P2X(1)R (IC(50), 0.3 microM) and at P2X(3)R (IC(50), 2.4 microM). 4 Phenolphthalein base was a potent irreversible antagonist at P2X(1)R (IC(50), 1 microM), whereas Phenolphthalein K(+)-salt was 25-fold less potent here. 5 Phenolphthalein base was a reversible antagonist of ATP responses at rat P2X(4)R (IC(50), 26 microM), whereas Phenolphthalein K(+)-salt was inactive. 6 Dimethyl sulphoxide (DMSO), used to dissolve lipophilic extracts, showed pharmacological activity by itself at rat P2X(1)R and P2X(4)R. 7 Thus, Phenol red and related compounds are antagonists at rat P2X(1)R, but are also active at other rat P2XRs. Phenolphthalein base is a newly identified, low potency antagonist of ATP responses at P2X(4)R. Culture media containing these red dyes should be used cautiously in future pharmacological studies of P2XRs. Also, wherever possible, the solvent DMSO should be used with caution. PMID- 15778740 TI - Involvement of reactive oxygen species in cyclic stretch-induced NF-kappaB activation in human fibroblast cells. AB - 1 Uniaxial cyclic stretch leads to an upregulation of cyclooxygenase (COX)-2 through increases in the intracellular Ca(2+) concentration via the stretch activated (SA) channel and following nuclear factor kappa B (NF-kappaB) activation in human fibroblasts. However, the signaling mechanism as to how the elevated Ca(2+) activates NF-kappaB is unknown. In this study, we examined the involvement of reactive oxygen species (ROS) as an intermediate signal, which links the elevated Ca(2+) with NF-kappaB activation. 2 4-Hydroxy-2-nonenal (HNE) was produced and modified IkappaB peaking at 2 min. The phosphorylation of IkappaB peaked at 8 min. HNE modification and IkappaB phosphorylation, NF-kappaB translocation to the nucleus, and following COX-2 production were inhibited by extracellular Ca(2+) removal or Gd(3+) application, as well as by the antioxidants. The stretch-induced Ca(2+) increase was inhibited by extracellular Ca(2+) removal, or Gd(3+) application. 3 IkappaB kinase (IKK) activity peaked at 4 min, which was inhibited by extracellular Ca(2+) removal, Gd(3+) or the antioxidants. IKK was also HNE-modified and, similarly to IkappaB, peaked at 2 min. IKK under static conditions was activated by exogenously applied HNE at a relatively low dose (1 microM), while it was inhibited at higher concentrations, suggesting that HNE could be one of the candidate signals in the stretch-induced NF-kappaB activation. 4 The present study suggests that the NF-kappaB activation by cyclic stretch is mediated by the following signal cascade: SA channel activation --> intracellular Ca(2+) increase --> production of ROS --> activation of IKK --> phosphorylation of IkappaB --> NF-kappaB translocation to the nucleus. PMID- 15778741 TI - Tachykinergic neurotransmission is enhanced in duodenum from dystrophic (mdx) mice. AB - 1 Duodenal longitudinal muscle of mdx mice, an animal model for Duchenne muscular dystrophy, showed a decrease in the electrically evoked nonadrenergic, noncholinergic (NANC) inhibitory responses associated with a reduction of the participation of nitric oxide (NO). In this study, we investigated whether the impairment of NO could also lead to alterations in the NANC excitatory transmission. 2 Nerve-evoked responses consisted of an inhibitory phase followed, at the end of stimulation, by an excitatory response characterised by an increase in amplitude of the spontaneous contractions. In mdx mice, the amplitude of the nerve-evoked contractions was significantly higher than in normals. 3 N(omega) nitro-L-arginine methyl ester (L-NAME) or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1 one (ODQ), an inhibitor of soluble guanylyl cyclase, increased the amplitude of the nerve-evoked contractions only in normals, being ineffective in mdx mice. Apamin, a blocker of Ca(2+)-dependent potassium channels, failed to affect the nerve-evoked contractions. 4 In both models, substance P and neurokinin A produced concentration-dependent contractions, reduced by tachykinin NK(1) and NK(2) receptor antagonists, respectively. Moreover, NK(1) and NK(2) receptor antagonists reduced the amplitude of the nerve-evoked contractions. 5 Sodium nitroprusside (SNP) reduced the amplitude of nerve-evoked contractions similarly in normal and mdx mice. ODQ, but not apamin, prevented the SNP-induced effects. SNP did not affect the contractions induced by exogenous tachykinins. 6 The results suggest that NO can exert an inhibitory modulatory role on tachykinergic excitatory transmission via activation of guanylyl cyclase in mouse duodenum. In mdx mice, the impairment of NO function leads to an increase in the nerve-evoked contractions. PMID- 15778742 TI - GW274150 and GW273629 are potent and highly selective inhibitors of inducible nitric oxide synthase in vitro and in vivo. AB - 1 GW274150 ([2-[(1-iminoethyl) amino]ethyl]-L-homocysteine) and GW273629 (3-[[2 [(1-iminoethyl)amino]ethyl]sulphonyl]-L-alanine) are potent, time-dependent, highly selective inhibitors of human inducible nitric oxide synthase (iNOS) vs endothelial NOS (eNOS) (>100-fold) or neuronal NOS (nNOS) (>80-fold). GW274150 and GW273629 are arginine competitive, NADPH-dependent inhibitors of human iNOS with steady state K(d) values of <40 and <90 nM, respectively. 2 GW274150 and GW273629 inhibited intracellular iNOS in J774 cells in a time-dependent manner, reaching IC(50) values of 0.2+/-0.04 and 1.3+/-0.16 microM, respectively. They were also acutely selective in intact rat tissues: GW274150 was >260-fold and 219 fold selective for iNOS against eNOS and nNOS, respectively, while GW273629 was >150-fold and 365-fold selective for iNOS against eNOS and nNOS, respectively. 3 The pharmacokinetic profile of GW274150 was biphasic in healthy rats and mice with a terminal half-life of approximately 6 h. That of GW273629 was also biphasic in rats, producing a terminal half-life of approximately 3 h. In mice however, elimination of GW273629 appeared monophasic and more rapid (approximately 10 min). Both compounds show a high oral bioavailability (>90%) in rats and mice. 4 GW274150 was effective in inhibiting LPS-induced plasma NO(x) levels in mice with an ED(50) of 3.2+/-0.7 mg kg(-1) after 14 h intraperitoneally (i.p.) and 3.8+/-1.5 mg kg(-1) after 14 h when administered orally. GW273629 showed shorter-lived effects on plasma NO(x) and an ED(50) of 9+/-2 mg kg(-1) after 2 h when administered i.p. 5 The effects of GW274150 and GW273629 in vivo were consistent with high selectivity for iNOS, as these inhibitors were of low potency against nNOS in the rat cerebellum and did not cause significant effects on blood pressure in instrumented mice. PMID- 15778744 TI - Estimating the impact of the next influenza pandemic on population health and health sector capacity in New Zealand. AB - AIM: To estimate the impact of the next influenza pandemic on population health and health sector capacity in New Zealand. METHOD: Population data for New Zealand was used with the software package 'FluAid' (CDC, Atlanta). Additional data was used to provide estimates of impacts on health sector capacity. RESULTS: For incidence rates in the 15% to 35% range for the first pandemic wave, the modelling results give a range of 1600 to 3700 deaths attributable to pandemic influenza. The estimated range of hospitalisations was between 6900 and 16,200. The estimated number of cases of illness requiring medical consultation ranged from 325,000 to 759,000. For the peak week of an 8-week epidemic (35% incidence scenario), it was estimated that 42% of all public hospital beds would be required at least for some proportion of the week and that the average general practitioner would be consulted by around 80 people with influenza. CONCLUSION: This modelling work has a number of limitations and so these results could still substantially over- or under-estimate the impact of the next influenza pandemic. Nevertheless, the potentially severe impact of pandemic influenza on population health and health sector capacity provides a strong case for health authorities to intensify preparatory efforts and to strengthen health sector infrastructure. PMID- 15778743 TI - CB1 cannabinoid receptor-mediated modulation of food intake in mice. AB - 1 Marijuana's appetite-increasing effects have long been known. Recent research suggests that the CB(1) cannabinoid receptor antagonist SR141716A may suppress appetite. This study represents a further, systematic investigation of the role of CB(1) cannabinoid receptors in the pharmacological effects of cannabinoids on food intake. 2 Mice were food-restricted for 24 h and then allowed access to their regular rodent chow for 1 h. Whereas the CB(1) antagonist SR141716A dose dependently decreased food consumption at doses that did not affect motor activity, Delta(9)-tetrahydrocannabinol (Delta(9)-THC) increased food consumption at doses that had no effect on motor activity. O-3259 and O-3257, structural analogs of SR141716A, produced effects similar to those of the parent compound. 3 Amphetamine (a known anorectic) and diazepam (a benzodiazepine and CNS depressant) decreased food consumption, but only at doses that also increased or decreased motor activity, respectively. The CB(2) cannabinoid receptor antagonist SR144528 and the nonpsychoactive cannabinoid cannabidiol did not affect food intake nor activity. 4 SR141716A decreased feeding in wild-type mice, but lacked pharmacological activity in CB(1) knockout mice; however, basal food intake was lower in CB(1) knockout mice. Amphetamine decreased feeding in both mouse genotypes. 5 These results suggest that SR141716A may affect the actions of endogenous cannabinoids in regulating appetite or that it may have effects of its own aside from antagonism of cannabinoid effects (e.g., decreased feeding behavior and locomotor stimulation). In either case, these results strongly suggest that CB(1) receptors may play a role in regulation of feeding behavior. PMID- 15778745 TI - New Zealand's preparedness for the next influenza pandemic. PMID- 15778746 TI - Screening for chronic hepatitis B infection in New Zealand: unfinished business. PMID- 15778747 TI - The New Zealand Hepatitis B Screening Programme: screening coverage and prevalence of chronic hepatitis B infection. AB - AIM: To report on screening coverage and the distribution of HBsAg (a marker of chronic hepatitis B virus infection) among participants in the New Zealand Hepatitis B Screening Programme. METHOD: Coverage and crude and age-standardised prevalence rates of HBsAg by age group, sex, ethnic group, and region were calculated from data held by the two providers and the New Zealand 2001 Census. RESULTS: 177,000 people were tested for hepatitis B virus infection (51% of the programme targets and 27% of Census 2001 eligible population), with highest coverage among women (28.9%) and Pacific people (34.9%). Overall, 5.7% (10,176) of participants were HBsAg-positive and there were significant regional, ethnic group, and gender differences. 5.6% of Maori, 7.3% of Pacific people, and 6.2% of Asians were HBsAg-positive, and men were more likely to test HBsAg-positive (6.1%) than women (5.4%). CONCLUSIONS: Previous estimates of HBsAg prevalence among Maori and Pacific people from smaller surveys were confirmed and new information obtained about the distribution of hepatitis B virus infection among Pacific Islands and Asian populations in New Zealand. Opportunistic screening of adults in these populations should continue in order to identify others with (as yet undetected) infection. Regular follow-up of people with chronic hepatitis B virus infection should also continue. Ongoing outcome monitoring is now needed to judge whether this unique programme has been an effective component of New Zealand's hepatitis B control strategy and whether it is a worthwhile investment of resources. PMID- 15778748 TI - Sexually transmitted infections in New Zealand in 2003. AB - AIMS: To describe the current burden and trends of sexually transmitted infections (STIs) in New Zealand (NZ) since 1999, as reported by current surveillance methods. METHODS: Clinic rates were calculated by dividing the number of diagnoses by the total number of clinic visits. Laboratory rates were calculated using the NZ Census 2001 population data for the Auckland, Waikato, and Bay of Plenty regions. RESULTS: In 2003, chlamydia was the most commonly diagnosed STI in sexual health (SHCs) and family planning clinics (FPCs), followed by genital warts. Laboratory surveillance reported a chlamydia rate of 653.0 per 100,000 population and a gonorrhoea rate of 90.2 per 100,000 population. The highest rates of chlamydia and gonorrhoea were in the 15 to 19 years age group. From 2002 to 2003, both chlamydia and gonorrhoea cases have increased by 14.0% in SHCs. In FPCs, chlamydia increased by 25.9% and gonorrhoea increased by 11.4%. Since 2002, numbers of chlamydia and gonorrhoea cases have increased by 14.0% in SHCs and by 25.9% and 11.4%, respectively, in FPCs. Maori and Pacific Peoples continue to be disproportionately affected by STIs. CONCLUSIONS: Current national surveillance methods are unrepresentative of the NZ population and do not provide accurate estimates of the population burden of STIs. Expansion of laboratory surveillance (to accurately reflect all areas of NZ) is needed and is currently under active consideration. PMID- 15778749 TI - High rates of chlamydia in patients referred for termination of pregnancy: treatment, contact tracing, and implications for screening. AB - AIMS: To determine the rate of chlamydia and other sexually transmitted infections (STIs), and to describe treatment and factors associated with chlamydia in patients presenting for a termination of pregnancy (TOP). METHODS: A retrospective audit of patients attending one of two TOP clinics from 1 February 2003. (Clinic A, n=500; Clinic B, n=501). Age, ethnicity, marital status, previous pregnancies, contraception, STIs, and treatment were recorded. RESULTS: Ten percent of patients tested positive for an STI. Chlamydia was most commonly detected, in 7.7% of all patients. Higher rates of chlamydia were observed at clinic B (10.2% vs 5.2%, p=0.005) and in under 25 year olds (11.2% vs 3.6%, p<0.001). Rates of chlamydia in Pacific women were 18.6%, in Maori 12.9%, in Asian 7.3% and 4.4% in New Zealand European women. All patients testing positive for chlamydia were treated prior to TOP but only 41% of partners were treated. Other infections detected included 18 cases of human papillomavirus (HPV), three cases of trichomoniasis, one case of gonorrhoea, and one case of syphilis. CONCLUSIONS: There is a high rate of chlamydia in women presenting for TOP, particularly in under 25 year olds, Pacific, and Maori women. There is an immediate need for policymakers to respond to this increasing burden of chlamydia by instigating targeted education, guidelines, and mandatory chlamydia screening and contact tracing for pregnant women. PMID- 15778750 TI - Is syphilis resurgent in New Zealand in the 21st century? A case series of infectious syphilis presenting to the Auckland Sexual Health Service. AB - Bacterial sexually transmitted infections such as chlamydia and gonorrhoea have been on the rise in recent years in New Zealand. Infectious syphilis has however remained rare over the last few decades, except in New Zealanders who have had sex overseas or in recent immigrants. However, in the previous 2 and a half years (1/2002-9/2004), the number of people presenting to Auckland Sexual Health Service with infectious syphilis has more than doubled. The main people at risk for acquiring infectious syphilis appear to be either men who have sex with men, or heterosexuals who have recently had sex overseas. More information is urgently required about the epidemiology of syphilis in New Zealand. One expedient way to determine whether we have an emerging epidemic would be to make syphilis a notifiable condition. PMID- 15778751 TI - Findings and outcome of teenage women referred for colposcopy at Christchurch Women's Hospital, New Zealand. AB - AIM: To determine the colposcopic findings, treatment, and follow up of 15-19 year old women referred to a large public colposcopy clinic. METHOD: A retrospective review of the colposcopy database and clinical records between 1995 and 2001. RESULTS: 243 women or 4.7% of new referrals were aged 15-19 years. Referral smears were high-grade squamous lesions (HGSL) in 15%, and low-grade squamous lesions (LGSL) or atypical squamous cells of uncertain significance (ASCUS) in 82%. Following colposcopy, 21% had biopsy proven high-grade abnormalities but only 4% had grade 3 cervical intraepithelial neoplasia (CIN3). Of those with LGSL or ASCUS smears, 2% had biopsy-proven CIN3; and of those with HGSL smears, 14% had biopsy proven CIN3. Women underwent a mean of 4.1 colposcopy sessions; 67% underwent treatment and 26% were discharged without treatment. Sixteen percent of the women were lost to follow-up. Treatment failure occurred in 8% of those treated. For women undergoing observation, 25% were discharged after the first follow-up and less than 3% progressed to CIN3. CONCLUSIONS: The rate of CIN3 in women under the age of 20 referred to colposcopy at Christchurch Women's Hospital is low. Screening teenage women results in invasive investigation and treatment without proven benefit and is not recommended. A conservative approach to the management of young women with low-grade squamous intraepithelial abnormalities is also advocated. PMID- 15778752 TI - The utility of blood cultures in the management of non-facial cellulitis appears to be low. AB - AIM: To assess the utility of blood cultures in the management of patients presenting to the Emergency Department at Christchurch Hospital from the community with non-facial cellulitis (or soft tissue infection) and no other morbidity. METHODS: A multidisciplinary team formulated the search protocol. A systematic review methodology was used. Seven electronic databases were searched for clinical studies of blood culture utility in patients with non-facial cellulitis. Relevant studies were appraised using predetermined validity assessment criteria. Conclusions were presented based on an assessment of the validity and applicability of the evidence. RESULTS: Seventeen studies were identified as addressing the topic at least as part of a secondary objective for the study. All were retrospective reviews or case series and were often associated with significant methodological limitations. CONCLUSIONS: Blood cultures are rarely positive in patients presenting from the community with non facial cellulitis. When they are positive, initial empiric therapy is usually adequate to treat pathogenic bacteria. The available evidence does not support the routine use of blood cultures in the clinical management of healthy adults presenting with non-facial cellulitis at the Emergency Department. PMID- 15778754 TI - Avoiding liability under the Commerce Act--what can be learned from the Ophthalmologists' case? AB - AIM: To consider the application of the Commerce Act 1986 to the conduct of medical practitioners and practitioner associations and societies, with special reference to the recent Ophthalmologists' case and the lessons which can be learned from that. DISCUSSION: The Commerce Commission recently took successful action (under Section 27 [s27] of the Commerce Act) against the Ophthalmological Society of New Zealand and several ophthalmologists from the lower South Island. The case illustrates the ease with which practitioners can be drawn into anticompetitive arrangements, and practitioner associations and societies can become liable under the Act through the conduct of their officeholders. The article discusses the relevant prohibitions under the Act, and the way in which these can relate to individual practitioners and practitioner associations and societies. In particular, s27 applies not only when the purpose of an arrangement substantially lessens competition, but also when the effect or likely effect of the arrangement substantially lessens competition. Therefore arguing that conduct is for ethical or safety reasons will not be sufficient to avoid liability. CONCLUSION: Markets for medical services are treated just like the markets for most other services under the Commerce Act. Individual practitioners should avoid becoming involved in or giving support to conduct or arrangements, which may be anticompetitive. Practitioner associations and societies should exercise control over the actions of their officeholders, especially where the purpose or effect of these could be considered to be anticompetitive. PMID- 15778753 TI - Demographic variation in the use of antibiotics in a New Zealand town. AB - AIMS: To describe the use of antibiotics in one New Zealand town, and to investigate relationships between antibiotic use and gender, age, and socioeconomic status. METHODS: Data from dispensing computers in all community pharmacies in the town were extracted. All dispensings of antibiotics to residents in the town were identified. Discrete individuals were identified and, where possible, linked with data on gender, age and the socioeconomic status (NZDep) of the area in which they lived. RESULTS: 42% of residents in the town received one or more dispensing of antibiotics in 2002. These people, on average, received 2.1 dispensings. Children received antibiotics more often than adults, females more than males and there was a strong relationship between socioeconomic status and antibiotic dispensings. CONCLUSIONS: Rates of antibiotic use in the community are strongly influenced by age, gender and socioeconomic status. PMID- 15778755 TI - Mycobacterium fortuitum infection caused by a cat bite. PMID- 15778756 TI - Idiopathic granulomatous hypophysitis. PMID- 15778757 TI - Tuberculosis in Auckland autopsies, revisited. AB - AIMS: To review the cases of tuberculosis found at autopsy in the Auckland Coronial Autopsy Service in the previous 10 years, and compare the results with that of a similar study for the period 1975 to 1992. Cases which were not diagnosed prior to autopsy are scrutinised as to why the diagnosis may have been missed. METHODS: A computerised search of the Forensic Pathology Department database at Auckland Hospital for cases of tuberculosis or atypical mycobacterial infection found at autopsy was done. Basic demographic data, past medical history, and police reports were analysed. The cases were placed into five groups for analysis. RESULTS: A total of 30 cases of tuberculosis (including 3 cases of atypical mycobacterial infection) were found. A large proportion (70%) was undiagnosed before death, and each case was potentially infective. Two cases had respiratory symptoms suggestive of tuberculosis, and a further 5 cases had generalised symptoms which were unexplained yet tuberculosis was not considered. CONCLUSIONS: In those cases that were missed, the autopsy played a vital role by identifying the previously undiagnosed cases of tuberculosis and ensuring contact screening. Awareness of tuberculosis and its increasing prevalence in New Zealand is essential for minimising missed diagnoses. PMID- 15778758 TI - COX-2 inhibitors--first, do no harm. PMID- 15778759 TI - Cord blood banking. PMID- 15778760 TI - Investigation for iron deficiency anaemia. PMID- 15778761 TI - Opioid poisoning deaths in New Zealand. PMID- 15778762 TI - Quality use of medicines activities. PMID- 15778763 TI - At last, some good news. PMID- 15778764 TI - Electron-conducting quantum dot solids: novel materials based on colloidal semiconductor nanocrystals. AB - We review the optical and electrical properties of solids that are composed of semiconductor nanocrystals. Crystals, with dimensions in the nanometre range, of II-VI, IV-VI and III-V compound semiconductors, can be prepared by wet-chemical methods with a remarkable control of their size and shape, and surface chemistry. In the uncharged ground state, such nanocrystals are insulators. Electrons can be added, one by one, to the conduction orbitals, forming artificial atoms strongly confined in the nanocrystal. Semiconductor nanocrystals form the building blocks for larger architectures, which self-assemble due to van der Waals interactions. The electronic structure of the quantum dot solids prepared in such a way is determined by the orbital set of the nanocrystal building blocks and the electronic coupling between them. The opto-electronic properties are dramatically altered by electron injection into the orbitals. We discuss the optical and electrical properties of quantum dot solids in which the electron occupation of the orbitals is controlled by the electrochemical potential. PMID- 15778765 TI - Continuous chirality measures in transition metal chemistry. AB - The definition of the continuous chirality measure(CCM) is provided and its applications are summarized in this tutorial review, with special emphasis on the field of transition metal complexes. The CCM approach, developed in recent years, provides a quantitative parameter that evaluates the degree of chirality of a given molecule. Many quantitative structural correlations with chirality have been identified for most of the important families of metal complexes. Our recent research has shown that one can associate the chirality measures with, e.g., enantioselectivity in asymmetric catalysis. We also explore a fragment approach to chirality in which we investigate which part of a molecule is responsible for the chirality-associated properties of a given family of compounds. PMID- 15778766 TI - Electron and energy transfer modulation with photochromic switches. AB - This tutorial review illustrates how work on the reversible interconversion between the colorless and colored forms of photochromic compounds can be exploited to modulate electron and energy transfer processes. Indeed, a photochrome can be designed to accept electrons or energy from a complementary donor in one of its two states only. Alternatively, the photoinduced transformations associated with a photochromic switch can be engineered to control the relative orientation and distance of donor-acceptor pairs. If either the donor or the acceptor is fluorescent, the photoregulated transfer of energy or electrons results in the modulation of the emission intensity. Thus, these fascinating molecular and supramolecular systems can advance the basic understanding of electron and energy transfer processes, while leading to viable operating principles to control light with light. PMID- 15778767 TI - Artificial metalloenzymes: proteins as hosts for enantioselective catalysis. AB - Enantioselective catalysis is one of the most efficient ways to synthesize high added-value enantiomerically pure organic compounds. As the subtle details which govern enantioselection cannot be reliably predicted or computed, catalysis relies more and more on a combinatorial approach. Biocatalysis offers an attractive, and often complementary, alternative for the synthesis of enantiopure products. From a combinatorial perspective, the potential of directed evolution techniques in optimizing an enzyme's selectivity is unrivaled. In this review, attention is focused on the construction of artificial metalloenzymes for enantioselective catalytic applications. Such systems are shown to combine properties of both homogeneous and enzymatic kingdoms. This review also includes our recent research results and implications in the development of new semisynthetic metalloproteins for the enantioselective hydrogenation of N protected dehydro-amino acids. PMID- 15778768 TI - Calorimetric and computational study of sulfur-containing six-membered rings. AB - Thermochemical data, and in particular the enthalpies of formation of oxygen- and sulfur-containing six-membered heterocycles provide essential information on the factors responsible for the contrasting behavior (structural, conformational and reactivity) between these types of compounds. A proper understanding of the experimental observations requires theoretical modeling in order to confirm the relative importance of the steric, electronic, electrostatic and stereoelectronic interactions that are responsible of the enthalpies of formation for the heterocyclic compounds of interest. PMID- 15778769 TI - Anti-inflammatory metabolites from marine sponges. AB - Marine sponges are a rich source of biologically active secondary metabolites with novel chemical structures. Eighty four anti-inflammatory compounds have been isolated from marine sponges. This is the first comprehensive review presenting the structures and anti-inflammatory activities of marine sponge metabolites. (100 references). PMID- 15778770 TI - Carotid stenosis and carotid plaque analysis relevant to carotid endarterectomy and stent-assisted angioplasty. AB - The primary objective of this cadaveric study was to review the morphological variations of the anatomy of the human carotid artery bifurcation relevant to carotid endarterectomy (CEA) and carotid artery stent-supported angioplasty (CSSA). We quantify carotid bifurcation plaque morphology. Results showed that the angle of deviation at the origin of the internal carotid artery (ICA), in relation to the common carotid artery (CCA), measured a mean of 21.8 degrees with a range from seven to 45 degrees. This anatomical finding is important for the interventionalist concerned with insertion of a carotid stent. The angle of the ICA origin may be an independent risk factor for early atherosclerotic changes at the ICA bulb. Carotid bifurcation plaque was observed in a small, random cohort of seven out of 13 cadavers, and contributed to a mean stenosis of 15.2% (range 5.0-34.8%). Plaque morphology (n = 7) showed haemorrhage (29%), superficial thrombosis (57%), calcification (71%), areas of focal necrosis (71%), neovascularisation (14%) and infiltrates (29%). Ulcerations were not detected. Although four out of 13 patients (31%) died of a cerebrovascular accident, the cause of cerebral apoplexy was thought not to be associated with the carotid bifurcation pathology. 'Re-boring' of occluding plaque, as in CEA, offers potential volumetric anatomical advantage over CSSA within the carotid bifurcation and bulb. In conclusion, precise and applied knowledge of carotid bifurcation anatomy is critical to reduce technical complications during CEA or CSSA. This information may reduce potential dangers of iatrogenic thrombo embolism and ensuing neurologic deficits. Patients with low-grade carotid stenosis, evidence of focal plaque necrosis, are at risk of spontaneous plaque cap rupture, distal thromboembolism and stroke. PMID- 15778771 TI - Proposed mechanisms for the anabolic steroid-induced increase in myocardial susceptibility to ischaemia/reperfusion injury. AB - Androgenic anabolic steroids (AAS) are often used by athletes to enhance athletic performance but are strongly associated with detrimental cardiovascular effects including sudden cardiac death. HYPOTHESIS: AAS use increases myocardial susceptibility to ischaemia/reperfusion injury. METHODS: Rats were trained (swimming) with or without intramuscular injection of nandrolone laurate (0.375 mg/kg). Untrained rats with or without nandrolone served as controls. Hearts were mounted on the Langendorff perfusion apparatus and mechanical function was measured before and after 20-min normothermic global ischaemia. Myocardial tissue samples were collected for determination of tissue cyclic nucleotide and TNFalpha concentrations. RESULTS: Anabolic steroids decreased the rate pressure product (RPP) of the exercise-trained rat heart [34 582 +/- 1 778 mmHg/min vs 28 868 +/- 2 446 mmHg/min for exercise-trained steroid-treated hearts (p < 0.05)]. Reperfusion RPP was lower in both the sedentary, and the exercise-trained, steroid-treated hearts than in their concurrent vehicle-treated controls (18 276 +/- 2 026 mmHg/min vs 12 018 +/- 1 725 mmHg/min for sedentary steroid-treated hearts and, 21 892 +/- 2 912 mmHg vs 12 838 +/- 1 536 mmHg/min for exercise trained steroid-treated hearts). Myocardial TNFalpha [267.75 +/- 44.25 pg/g vs 190.00 +/- 15.75 pg/g (p < 0.05)] and cAMP concentrations [406.04 +/- 18.41 pmol/g vs 235.6 +/- 43.26 pmol/g (p < 0.05)] were elevated in the steroid-treated hearts when compared with their untreated counterparts. CONCLUSIONS: Supraphysiological doses of anabolic steroids, whether taken during exercise training or under sedentary conditions increase myocardial susceptibility to ischaemia/reperfusion injury in our model. This increased susceptibility may be related to steroid-induced increases in the pre-ischaemic myocardial cAMP concentrations and/or increases in both pre-ischaemic and reperfusion TNFalpha concentrations. PMID- 15778772 TI - Predictors of the successful outcome of one-year survivors of coronary artery bypass surgery. AB - This study was conducted on a South African cohort to establish the actors that may predict the successful outcome of coronary artery bypass surgery when assessed in terms of improved quality of life one year after the surgery. Information was sought on the socio-economic status of patients, their risk factor profiles and clinical history. From the patient files, information was recorded on left ventricular ejection fraction, number of vessels bypassed, bypass time, and aortic cross-clamp time. The characteristics that were predictive of a successful outcome one year after surgery were identified. Patients in this study represented a high-risk population with multiple risk factors (obese, heavy smokers, hypertensive, hypercholesterolaemic, inactive, family history of heart disease, diabetes, and regular intake of alcohol). Measured medical parameters could not distinguish between the group with an improved quality of life and the group who did not have improved quality of life. One year after CABG all patients with an improved quality of life were men. The additional identified predictor variables for a successful outcome were: being married, patients' height, the knowledge that smoking affects the cardiovascular system, number of years that sporting activities were stopped prior to CABG surgery, a better quality sex life after the operation, acceptance of self responsibility for rehabilitation, and the spouse knowing the diet the patient should follow. The predictors of a successful outcome at the time of the operation were: being married (OR = 22.6; p = 0.02); taller than 170 cm (OR = 15.5; p = 0.01); stopped all sporting activities for a period less than 20 years prior to their surgery (OR 11.4; p = 0.01). We concluded that the outcome of coronary artery bypass surgery could not be predicted on the basis of a medical model that considers exclusively the extent of the patient's disease and associated co-morbidities. Patients should be carefully selected and an intensive post-operative educational intervention should be provided to patients and their spouses/caregivers. PMID- 15778773 TI - Endovascular aortic aneurysm repair by a multidisciplinary team: lessons learned and six-year clinical update. AB - BACKGROUND: Endovascular aneurysm repair (EVAR) (using an Federal Drug Association-approved AneuRx device) compared to conventional surgical repair of abdominal aortic aneurysm (AAA) previously rendered favourable outcomes regarding post-operative pain, avoidance of laparotomy, and rapid rehabilitation and hospital discharge in high-risk patients, including octagenarians. OBJECTIVES: To assess the safety, reduction in aneurysm-related deaths, and interim survival data up to 72 months after AAA exclusion by endoluminal endografts (EVAR). DESIGN: We carried out an open, controlled, prospective, multidisciplinary EVAR study for the period 1998 to 2003 (six years). In the earlier part of the study, EVAR was compared with previously published results of conventional open aneurysmectomy surgery. SETTING: Heart Unit, Panorama Medi-Clinic, Parow, South Africa. PARTICIPANTS: We recruited adult male and female patients presenting with AAA and fulfilling the inclusion criteria for endovascular repair, as recommended by the consensus 2003 meeting of the Vascular Association of South Africa (VASSA). All patients were offered open surgery as an alternative and were entered into the VASSA EVAR trial registry. Pre-operatively, AAA anatomy was assessed by spiral-computed tomography (CT), and selectively with conventional angiography and intravascular ultrasound (IVUS). Informed consent was obtained in accordance with the recommendations of the Senate of Surgery Paper 2, Ethical Guidelines, Great Britain and Ireland. Patients underwent EVAR by a multidisciplinary interventional team. INTERVENTIONS: Two hundred and seven adult patients with AAA were assessed. Forty-four of the 207 (21.2%) were excluded from EVAR because of irreversible comorbid factors and complex aneurysm morphology. One hundred and sixty-three patients (78%), with a mean age of 70.7 years (range 60-91 years), underwent EVAR (1998-2003). Five patients were lost to follow-up (3%). Median AAA diameter was 56.9 mm and ASA ratings were I, 1.2%; II, 15.9%; III, 57%; IV, 22.6%; and V, 2.4%. EVAR was performed in high- and low-risk categories of both sexes. Most patients were in ASA groups III and IV. DEVICES DEPLOYED: EVAR was performed using a selection of endografts over 72 months- AneuRx (Medtronic) 47; Talent (Medtronic) 49; Vanguard three; Zenith (Cook) one; Powerlink (Endologix) 62; and other, one. RESULTS: Thirty-day outcome: successful deployment 99%, primary stent patency 97%, surgical conversion 0.6%, procedural or intra-operative mortality 1.2%, 30-day mortality 4.3%, endoleaks 1.84%, and secondary intraprocedural endovascular interventions 24.5%. Perioperative mortality was 3.1% (one aneurysm related). One patient had suspected endograft infection. Late mortality was 21.4% (35 patients due to co-morbidities, and one was aneurysm related). Follow-up was a median of 28.3 months (range 1-69 months). In 163 patients, two persisting endoleaks (1.2%) were detected. Endotension was detected in 3/163 (1.8%) with average sac increase of 0.8 cm. Conversion to open surgery was needed in one patient (0.6%). Co-morbidities that contributed to late mortality included multi-organ failure, ischaemic heart disease (IHD), cardiomyopathy, renal failure, stroke and cancer. One procedural rupture was fatal (0.6%). Two late ruptures occurred; one was successfully endostented and the other patient died after a failed surgical intervention (0.6%). Endovascular repair of AAA is more expensive than conventional surgery. Introduction of the Endologix stent has reduced operative time from 120 to 60 minutes in un complicated patients. Newer-generation aortic stents allow better control of negative remodeling and stent migration. CONCLUSION: A multidisciplinary team can safely perform EVAR, with a low 30-day mortality rate in selected patients graded ASA II-IV and with favourable aortic aneurysm morphology. About 22% of patients with AAA are not suited for EVAR. Persisting late endoleaks occurred in 1.2% of the cohort study and were not device specific. Life-long follow-up post EVAR is a prerequisite to detect late device failure, endoleaks and aneurysm-sac enlargement, and to assure the durability of these midterm results. Short-term aneurysm rupture prevention is a predictable outcome in high-risk groups. PMID- 15778774 TI - Ventricular tachycardia as an adverse effect of the African potato (Hypoxis sp.). PMID- 15778775 TI - Eprosartan (Teveten) offers new opportunities to prevent a second stroke. PMID- 15778776 TI - Interpreting the ACTION study - benefits in patients with stable angina and raised blood pressure. PMID- 15778777 TI - Pre-hospital use of aspirin in emergency setting - seldom with adverse events. PMID- 15778778 TI - ARBITER-2 : evidence that targeting HDL cholesterol slows atherosclerosis. PMID- 15778779 TI - New amlodipine (Amloc) product available in South Africa. PMID- 15778780 TI - Major trial shows cardiovascular benefits using new drug combination with perindopril. PMID- 15778781 TI - Valsartan - first angiotensin II receptor blocker licensed for post-MI use. PMID- 15778782 TI - Achieving synergy and improvements in cholesterol lowering. PMID- 15778783 TI - Renewed interest in lovastatin following launch of Lovachol. PMID- 15778784 TI - Prevention of microalbuminuria in type 2 diabetics. PMID- 15778785 TI - PEACE - an important yet neutral study of low-risk patients with CAD. PMID- 15778787 TI - Bad health informatics can kill--is evaluation the answer? AB - OBJECTIVE: Health care is entering the age of information society. It is evident that the use of modern information and communication technology (ICT) offers tremendous opportunities to improve health care. However, there are also hazards associated with ICT in health care. We want to present an overview of typical hazards associated with ICT in health care, and to discuss how ICT evaluation can be a solution. METHODS: We analyze examples of failures and problems associated with ICT in health care. This collection is also made available on a website. RESULTS AND CONCLUSION: Systematic, continuous evaluation of quality and effects of ICT during the whole life cycle of ICT components seems to be one important approach to detect and prevent possible ICT hazards and failures, supporting a higher quality of patient care. However, empirical studies proving this assumption are needed. PMID- 15778788 TI - Demand-driven evolution of IT systems in healthcare--a case study for improving interdisciplinary processes. AB - OBJECTIVES: To analyze and to optimize interdisciplinary clinical processes, to introduce an IT-supported model for demand-driven system evolution in healthcare, and to demonstrate the feasibility of the approach for a clinical example and to present an evaluation. METHODS: System evolution and change management are viewed as two sides of the same coin, thus formal methods for process analysis and IT system evolution were embedded into a goal-oriented change management model. Based on a process model, a Failure Mode and Effects Analysis (FMEA) and a computer simulation were performed. A tool for rapid application development (RAD) was used to incrementally improve the healthcare information system according to newly arising needs. RESULTS: Each of the formal methods used contributed to the successful reorganization of the interdisciplinary clinical process. An evaluation demonstrated significant improvements. An integrated IT application was implemented to support the optimized process. CONCLUSIONS: Process improvement is feasible and effective when formal methods for process analysis and requirements specification are used in a reasonable and goal oriented way. It might be necessary to trade off costs and benefits or simplify a given method in the context of a particular project. As the same information is utilized in different tools, it is supposed that the efforts for process analysis, documentation and implementation of adapted applications could be reduced if different tools were integrated and based on a single coherent reference model for description of clinical processes. PMID- 15778789 TI - Modeling cancer registration processes with an enhanced activity diagram. AB - OBJECTIVES: Adequate instruments are needed to reflect the complexity of routine cancer registry operations properly in a business model. The activity diagram is a key instrument of the Unified Modeling Language (UML) for the modeling of business processes. The authors aim to improve descriptions of processes in cancer registration, as well as in other public health domains, through the enhancements of an activity diagram notation within the standard semantics of UML. METHODS: The authors introduced the practical approach to enhance a conventional UML activity diagram, complementing it with the following business process concepts: timeline, duration for individual activities, responsibilities for individual activities within swimlanes, and descriptive text. RESULTS: The authors used an enhanced activity diagram for modeling surveillance processes in the cancer registration domain. Specific example illustrates the use of an enhanced activity diagram to visualize a process of linking cancer registry records with external mortality files. CONCLUSIONS: Enhanced activity diagram allows for the addition of more business concepts to a single diagram and can improve descriptions of processes in cancer registration, as well as in other domains. Additional features of an enhanced activity diagram allow to advance the visualization of cancer registration processes. That, in turn, promotes the clarification of issues related to the process timeline, responsibilities for particular operations, and collaborations among process participants. Our first experiences in a cancer registry best practices development workshop setting support the usefulness of such an approach. PMID- 15778790 TI - Design and development of a mobile system for supporting emergency triage. AB - OBJECTIVES: Our objective was to design and develop a mobile clinical decision support system for emergency triage of different acute pain presentations. The system should interact with existing hospital information systems, run on mobile computing devices (handheld computers) and be suitable for operation in weak connectivity conditions (with unstable connections between mobile clients and a server). METHODS: The MET (Mobile Emergency Triage) system was designed following an extended client-server architecture. The client component, responsible for triage decision support, is built as a knowledge-based system, with domain ontology separated from generic problem solving methods and used for the automatic creation of a user interface. RESULTS: The MET system is well suited for operation in the Emergency Department of a hospital. The system's external interactions are managed by the server, while the MET clients, running on handheld computers are used by clinicians for collecting clinical data and supporting triage at the bedside. The functionality of the MET client is distributed into specialized modules, responsible for triaging specific types of acute pain presentations. The modules are stored on the server, and on request they can be transferred and executed on the mobile clients. The modular design provides for easy extension of the system's functionality. A clinical trial of the MET system validated the appropriateness of the system's design, and proved the usefulness and acceptance of the system in clinical practice. CONCLUSIONS: The MET system captures the necessary hospital data, allows for entry of patient information, and provides triage support. By operating on handheld computers, it fits into the regular emergency department workflow without introducing any hindrances or disruptions. It supports triage anytime and anywhere, directly at the point of care, and also can be used as an electronic patient chart, facilitating structured data collection. PMID- 15778791 TI - The International Partnership for Health Informatics Education: lessons learned from six years of experience. AB - OBJECTIVES: To inform the medical and health informatics community on the rational, goals, and the achievements of the International Partnership for Health Informatics Education--IPHIE, (I phi E), that was established at six universities in 1999. METHODS: We elaborate on the overall goals of I phi E and describe the current state of affairs: the activities undertaken and faculty and student experience related to these activities. In addition we outline the lessons we have learned over these past six years and our plans for the future. RESULTS: I phi E members first started to collaborate by supporting and encouraging the exchange of talented students and faculty and by establishing joint master classes for honors students. Following the success of these activities, new initiatives were undertaken such as the organization of student workshops at medical informatics conferences and a joint course on strategic information management in hospitals in Europe. CONCLUSIONS: International partnerships such as I phi E take time to establish, and, if they are to be successful, maintaining leadership continuity is critically important. We are convinced that I phi E promotes professionalism of future medical informatics specialists. There will be a continuing growth of globalization in higher education. It will therefore become increasingly important to offer educational programs with international components. PMID- 15778792 TI - Automated tissue analysis--a bioinformatics perspective. AB - OBJECTIVES: Recent progress in automated tissue analysis (tissomics) provides reproducible phenotypical characterization of histological specimens. We introduce informatics tools to cluster and correlate quantitative tissue profiles with gene expression data. The great potential of synergies between tissue analysis and bioinformatics and its perspectives in medical research and computational diagnostics are discussed. METHODS: Key enablers in microscopic imaging and machine vision are reviewed to perform a high-throughput tissue analysis. Methodologies are described and results are demonstrated that support a combined analysis of tissue with gene expression profiles whereby the consideration of individual responses is key. RESULTS: Comprehensive histomorphometric profiles, extracted using machine vision, provide information regarding the components and heterogeneity of a tissue in a reproducible format amenable to data mining and analysis. Tissue quantitative information can be placed in synergetic context with bioinformatics data, such as gene expression profiles, for a more comprehensive stratification of individual responses. From a bioinformatics point of view tissue data are co-variants that support the identification of candidate genes relevant in tissue injury or disease. CONCLUSIONS: Progress in automated analytics enables the generation of quantitative data about tissue previously limited to visual histopathology. Such reproducible data sets can be statistically correlated and clustered throughout the continuum of bioinformatics. The combined approach supports a system-wide view of biology and has a potential to accelerate developments for a personalized computational diagnosis. PMID- 15778793 TI - Applying informatics in tissue engineering. AB - OBJECTIVE: To facilitate tissue engineering strategies determination with informatics tools. METHODS: Firstly, tissue engineering experimental data were standardized and integrated into a centralized database; secondly, we used data mining tools (e.g. artificial neural networks and decision trees) to predict the outcomes of tissue engineering strategies; thirdly, a strategy design algorithm was developed, and its efficacy was validated with animal experiments; lastly, we constructed an online database and a decision support system for tissue engineering. RESULTS: The artificial neural networks and the decision trees respectively predicted the outcomes of tissue engineering strategies with the predictive accuracy of 95.14% and 85.26%. Following the strategies generated by computer, we cured 18 of the 20 experimental animals with a significantly lower cost than usual. CONCLUSION: Informatics is beneficial for realizing safe, effective and economical tissue engineering. PMID- 15778794 TI - An inventory of evaluation studies of information technology in health care trends in evaluation research 1982-2002. AB - OBJECTIVES: During the last years the significance of evaluation studies as well as the interest in adequate methods and approaches for evaluation has grown in medical informatics. In order to put this discussion into historical perspective of evaluation research, we conducted a systematic review on trends in evaluation research of information technology in health care from 1982 to 2002. METHODS: The inventory is based on a systematic literature search in PubMed. Abstracts were included when they described an evaluation study of a computer-based component in health care. We identified 1035 papers from 1982 to 2002 and indexed them based on a multi-axial classification describing type of information system, study location, evaluation strategy, evaluation methods, evaluation setting, and evaluation focus. RESULTS AND CONCLUSIONS: We found interesting developments in evaluation research in the last 20 years. For example, there has been a strong shift from medical journals to medical informatics journals. With regard to methods, explanatory research and quantitative methods have dominated evaluation studies in the last 20 years. Since 1982, the number of lab studies and technical evaluation studies has declined, while the number of studies focusing on the influence of information technology on quality of care processes or outcome of patient care has increased. We interpret this shift as a sign of maturation of evaluation research in medical informatics. PMID- 15778795 TI - A methodological framework for the conversion of procedure classifications. AB - OBJECTIVES: During the adaptation of the Australian Refined Diagnosis Related Groups for Germany mapping tables between procedure classifications were needed. The mapping between the German OPS-301 2.0 and the Australian MBS-Extended should transfer the Australian expertise by keeping a well-established terminology system. METHODS: A methodological framework for the development of mapping tables had been developed based on the model for representation of semantics provided by the European Committee of Standardization. Two approaches were used; the concept based approach from OPS-301 2.0 to MBS-Extended and the class-based approach the other way round. A conversion had to be identified between 23,160 classes of the OPS-301 2.0 and 6,328 classes of the MBS-Extended in two asymmetrical mapping tables. RESULTS: The class-based approach leads to a low number of 6,980 conversions but misses 82.6% of the classes of the OPS-301. Because of domain incongruencies and missing domain completeness of the OPS-301 2.0 for non operative procedures 15.7% of the MBS-Extended-classes remain without conversion. The concept-based approach leads to a slightly higher mean number of conversions per class of 1.35 in comparison to 1.31 with the class-based approach. But it was possible to find conversions for 99.5% of the OPS-301 2.0-classes. 16.3% of the DRG-relevant classes of the MBS-Extended were missed. CONCLUSIONS: The class based approach was not useful, because the MBS-Extended is significantly broader than the OPS-301 2.0. An external validation study for the direction OPS-301 2.0 to MBS-Extended revealed a satisfactory quality. The empirical and the reference based approach are important alternatives to the ones used in this project. There are clear criteria about the appropriate application area for the methodological approaches presented here. PMID- 15778796 TI - The EpiLink record linkage software: presentation and results of linkage test on cancer registry files. AB - OBJECTIVES: Record linkage, the process of bringing together separately compiled but related records from different databases, is essential in many areas of biomedical research. We developed a record linkage program (EpiLink), which employs a simple mathematical approach. We describe the program and present results obtained testing it in a linkage task. METHODS: EpiLink was designed to be flexible with user-friendly settings to tailor linkage and operating parameters to specific linkage tasks, and employ deterministic, probabilistic or sequential deterministic-probabilistic linkage strategies as required. The user can also standardize data format, examine linkage results and accept or discard them. We used EpiLink to link a subset of cases of the Lombardy Cancer Registry (20,724 records) with the Social Security file of the population (1,021,846 records) covered by the registry. The linkage strategy was deterministic, followed by several probabilistic linkage steps. RESULTS: Manual inspection of the results showed that EpiLink achieved 98.8% specificity and 96.5% sensitivity. CONCLUSIONS: EpiLink is a practical and accurate means of linking records from different databases that can be used by non-statisticians and is efficient in terms of human and financial resources. PMID- 15778797 TI - Decision analysis for the assessment of a record linkage procedure: application to a perinatal network. AB - OBJECTIVES: According to European legislation, we must develop computer software allowing the linkage of medical records previously rendered anonymous. Some of them, like AUTOMATCH, are used in daily practice either to gather medical files in epidemiologic studies or for clinical purpose. In the first situation, the aim is to avoid homonymous errors, and in the second one, synonymous errors. The objective of this work is to study the effect of different parameters (number of identification variables, phonetic treatments of names, direct or probabilistic linkage procedure) on the reliability of the linkage in order to determine which strategy is the best according to the purpose of the linkage. METHODS: The assessment of the Burgundy Perinatal Network requires the linking of discharge abstracts of mothers and neonates, collected in all the hospitals of the region. Those data are used to compare direct and probabilistic linkage, using different parameterization strategies. RESULTS: If the linkage has to be performed in real time, so that no validation of indecisions generated by probabilistic linkage is possible, probabilistic linkage using three variables without any phonetic treatment seems to be the most appropriate approach, combined with a direct linkage using four variables applied to non-conclusive links. If a validation of indecisions is possible in an epidemiological study, probabilistic linkage using five variables, with a phonetic treatment adapted to the local language has to be preferred. For medical purpose, it should be combined with a direct linkage with four or five variables. CONCLUSION: This paper reveals that the time and money available to manage indecision as well as the purpose of the linkage are of paramount importance for choosing a linkage strategy. PMID- 15778798 TI - AdaptFlow: protocol-based medical treatment using adaptive workflows. AB - OBJECTIVES: In many medical domains investigator-initiated clinical trials are used to introduce new treatments and hence act as implementations of guideline based therapies. Trial protocols contain detailed instructions to conduct the therapy and additionally specify reactions to exceptional situations (for instance an infection or a toxicity). To increase quality in health care and raise the number of patients treated according to trial protocols, a consultation system is needed that supports the handling of the complex trial therapy processes efficiently. Our objective was to design and evaluate a consultation system that should 1) observe the status of the therapies currently being applied, 2) offer automatic recognition of exceptional situations and appropriate decision support and 3) provide an automatic adaptation of affected therapy processes to handle exceptional situations. METHODS: We applied a hybrid approach that combines process support for the timely and efficient execution of the therapy processes as offered by workflow management systems with a knowledge and rule base and a mechanism for dynamic workflow adaptation to change running therapy processes if induced by changed patient condition. RESULTS AND CONCLUSIONS: This approach has been implemented in the AdaptFlow prototype. We performed several evaluation studies on the practicability of the approach and the usefulness of the system. These studies show that the AdaptFlow prototype offers adequate support for the execution of real-world investigator-initiated trial protocols and is able to handle a large number of exceptions. PMID- 15778799 TI - Selection of predictor variables for pneumonia using neural networks and genetic algorithms. AB - BACKGROUND: Artificial neural networks (ANN) can be used to select sets of predictor variable that incorporate nonlinear interactions between variables. We used a genetic algorithm, with selection based on maximizing network accuracy and minimizing network input-layer cardinality, to evolve parsimonious sets of variables for predicting community-acquired pneumonia among patients with respiratory complaints. METHODS: ANN were trained on data from 1044 patients in a training cohort, and were applied to 116 patients in a testing cohort. Chromosomes with binary genes representing input-layer variables were operated on by crossover recombination, mutation, and probabilistic selection based on a fitness function incorporating both network accuracy and input-layer cardinality. RESULTS: The genetic algorithm evolved best 10-variable sets that discriminated pneumonia in the training cohort (ROC areas, 0.838 for selection based on average cross entropy (ENT); 0.954 for selection based on ROC area (ROC)), and in the testing cohort (ROC areas, 0.847 for ENT selection; 0.963 for ROC selection), with no significant differences between cohorts. Best variable sets based on the genetic algorithm using ROC selection discriminated pneumonia more accurately than variable sets based on stepwise neural networks (ROC areas, 0.954 versus 0.879, p = 0.030), or stepwise logistic regression (ROC areas, 0.954 versus 0.830, p = 0.000). Variable sets of lower cardinalities were also evolved, which also accurately discriminated pneumonia. CONCLUSION: Variable sets derived using a genetic algorithm for neural networks accurately discriminated pneumonia from other respiratory conditions, and did so with greater accuracy than variables derived using stepwise neural networks or logistic regression in some cases. PMID- 15778800 TI - In vivo and simulation study of artificial ventilation effects on energetic variables in cardiosurgical patients. AB - OBJECTIVES: The analysis of energetic ventricular variable changes during artificial ventilation, obtained by numerical simulation was done. Twenty-one sets of hemodynamic parameters for eight cardiosurgical patients were used to estimate left and right stroke work. The data were collected for three methods of ventilation: conventional, lung-protective (with minute ventilation diminished by half) and high frequency ventilation (with frequency 5, 10, or 15 Hz). METHODS: The computer simulator (CARDIOSIM) of the cardiovascular system, was used as a tool to calculate values of energetic ventricular variables for conditions that corresponded to these during in vivo measurements. Different methods of ventilation caused differences of intrathoracic pressure, haemodynamic and finally energetic ventricular variables. The trends of these variable changes were the same in in vivo and simulation studies, in the whole range of intrathoracic pressure changes (Pt = 1.5-3.5 mmHg). RESULTS: As values of main hemodynamic variables like cardiac output or arterial, systemic and pulmonary pressures were very close in both studies. Cardiac index and left ventricular stroke work also differed less than 10% for all examined patients and computer simulation. In a case of right ventricular stroke work the difference between in vivo data and simulation was a bit greater than 10% for two of eight patients under study. CONCLUSIONS: Our comparative analysis proved that numerical simulation is a very useful tool to predict changes of main hemodynamic and energy-related ventricular variables caused by different levels of positive Pt. It means that it can help an anesthesiologist to choose an appropriate method of artificial ventilation for cardiosurgical patients. PMID- 15778801 TI - Propagation of EEG activity in the beta and gamma band during movement imagery in humans. AB - OBJECTIVES: The objective of the paper was the determination of electrical brain activity propagation in sensorimotor areas during hand movement imagery. METHODS: Right-hand and left-hand movement imagination was studied in three subjects. The 10-channel Multivariate Autoregressive Model (MVAR) was fitted to EEG signals recorded from subsets of electrodes overlying central and related brain areas. By means of the Short-time Directed Transfer Function (SDTF) the propagation of brain activity as a function of frequency and time was found. RESULTS: During imagery the relation between propagations in gamma and beta bands changed significantly for electrodes overlying sensorimotor areas, namely the increase in gamma was accompanied by the decrease in the beta band. CONCLUSIONS: The hypothesis was put forward that these kinds of changes in flow of electrical brain activity are connected with the specific information processing. PMID- 15778802 TI - Integrated software suite for magnetocardiographic data analysis--a proposal based on an interactive programming environment. AB - OBJECTIVES: This paper describes an integrated software suite (ISS) for the processing of magnetocardiographic (MCG) recordings obtained with super conducting multi-channel systems having different characteristics. We aimed to develop a highly flexible suite including toolboxes for current MCG applications, organized consistently with an open architecture that allows function integrations and upgrades with minimal modifications; the suite was designed for the compliance not only of physicists and engineers but also of physicians, who have a different professional profile and are accustomed to retrieve information in different ways. METHODS: The MCG-ISS was designed to work with all common graphical user interface operative systems. MATLAB was chosen as the interactive programming environment (IPE), and the software was developed to achieve usability, interactivity, reliability, modularity, expansibility, interoperability, adaptability and graphics style tailoring. Three users, already experienced in MCG data analysis, have intensively tested MCG-ISS for six months. A great amount of MCG data on normal subjects and patients was used to assess software performances in terms of user compliance and confidence and total analysis time. RESULTS: The proposed suite is an all-in-one analysis tool that succeeded in speeding MCG data analysis up to about 55% with respect to standard reference routines; it consequently enhanced analysis performance and user compliance. CONCLUSIONS: Those results, together with the MCG-ISS advantage of being independent on the acquisition system, suggest that software suites like the proposed one could uphold a wider diffusion of MCG as a diagnostic tool in the clinical setting. PMID- 15778803 TI - A ruler for interpreting diagnostic test results. AB - OBJECTIVES: Bayes' rule formalizes how the pre-test probability of having a condition of interest is changed by a diagnostic test result to yield the post test probability of having the condition. To simplify this calculation a geometric solution in form of a ruler is presented. METHODS: Using odds and the likelihood ratio of a test result in favor of having the condition of interest, Bayes' rule can succinctly be expressed as "the posttest odds equals the pre-test odds times the likelihood ratio". Taking logarithms of both sides yields an additive equation. RESULTS: The additive log odds equation can easily be solved geometrically. We propose a ruler made of two scales to be adjusted laterally. A different, widely used solution in form of a nomogram was published by Fagan. CONCLUSIONS: Whilst use of the nomogram seems more obvious, the ruler may be easier to operate in clinical practice since no straight edge is needed for precise reading. Moreover, the ruler yields more intuitive results because it shows the change in probability due to a given test result on the same scale. PMID- 15778804 TI - Meta-analysis: a unifying meta-likelihood approach framing unobserved heterogeneity, study covariates, publication bias, and study quality. AB - OBJECTIVES: This contribution provides a unifying concept for meta-analysis integrating the handling of unobserved heterogeneity, study covariates, publication bias and study quality. It is important to consider these issues simultaneously to avoid the occurrence of artifacts, and a method for doing so is suggested here. METHODS: The approach is based upon the meta-likelihood in combination with a general linear nonparametric mixed model, which lays the ground for all inferential conclusions suggested here. RESULTS: The concept is illustrated at hand of a meta-analysis investigating the relationship of hormone replacement therapy and breast cancer. The phenomenon of interest has been investigated in many studies for a considerable time and different results were reported. In 1992 a meta-analysis by Sillero-Arenas et al. concluded a small, but significant overall effect of 1.06 on the relative risk scale. Using the meta likelihood approach it is demonstrated here that this meta-analysis is due to considerable unobserved heterogeneity. Furthermore, it is shown that new methods are available to model this heterogeneity successfully. It is argued further to include available study covariates to explain this heterogeneity in the meta analysis at hand. CONCLUSIONS: The topic of HRT and breast cancer has again very recently become an issue of public debate, when results of a large trial investigating the health effects of hormone replacement therapy were published indicating an increased risk for breast cancer (risk ratio of 1.26). Using an adequate regression model in the previously published meta-analysis an adjusted estimate of effect of 1.14 can be given which is considerably higher than the one published in the meta-analysis of Sillero-Arenas et al. In summary, it is hoped that the method suggested here contributes further to a good meta-analytic practice in public health and clinical disciplines. PMID- 15778805 TI - [Anaesthesia in patients with Parkinson's disease]. AB - The neurodegenerative death of dopaminergic neurons of the pars compacta of the substantia nigra leads to the classical triad of resting tremor, muscle rigidity, and bradykinesia of Parkinson's disease. Parkinson's disease is a common disease of elderly patients requiring perioperative anaesthesia. Particular anaesthetic problems are neurological, respiratory, and cardiovascular. The clinical features and the interaction of common anaesthetics with the drug therapy of the patient present an anaesthetic challenge and directly influence perioperative morbidity and mortality. PMID- 15778806 TI - [Pre-operative measurement of heart rate variability in diabetics: a method to estimate blood pressure stability during anaesthesia induction]. AB - BACKGROUND: In diabetics severe hyopotension can occur during anaesthesia as a consequence of cardiovascular autonomic neuropathy (CAN). A simple approach to recognize CAN is heart rate variability (HRV) measured at rest. The aim of this study was to determine the relationship between pre-operatively measured HRV and blood pressure stability during induction phase of anaesthesia. PATIENTS AND METHODS: A total of 35 diabetics undergoing an ophthalmosurgical procedure under general anaesthesia were investigated. HRV was examined one day before surgery by the following parameters: coefficient of variation (CV), root mean squared successive difference (RMSSD), and power spectrum. Anaesthesia was induced with thiopental and fentanyl and maintained with enflurane/N(2)O; tracheal intubation was performed after relaxation with vecuronium. Patients who developed a drop in systolic blood pressure below 90 mmHg during anaesthesia induction were assigned to group H (hypotensive), the other patients to group N (normotensive). The groups were compared regarding HRV and other variables. RESULTS: Of all patients 13 developed hypotension during anaesthesia induction (group H). The groups were comparable regarding the demographic data. Parameters of HRV, with the exception of spectral power in low frequency (LF) band of power spectrum, were significantly lower in group H. The groups differed mainly in relation to spectral power in the mid-frequency (MF) band of the power spectrum, and especially regarding CV. In patients with normal CV incidence of hypotension after anaesthesia induction was 11%, but in patients with abnormally reduced CV, 65% (p=0.002). CONCLUSION: The results confirm a significant relationship between HRV pre-operatively measured at rest and blood pressure stability during anaesthesia induction in diabetics. Particularly examination of CV, a simple test feasible within few minutes, may be useful in pre-operative risk stratification of these patients. Application of the time consuming traditional test combination to identify CAN seems to be unnecessary. PMID- 15778807 TI - Single-nucleotide polymorphisms in the C-reactive protein (CRP) gene promoter that affect transcription factor binding, alter transcriptional activity, and associate with differences in baseline serum CRP level. AB - To investigate whether functional polymorphisms exist in the C-reactive protein (CRP) gene, i.e., ones that contribute directly to differences in baseline CRP among individuals, we sequenced a 1,156-nucleotide-long stretch of the CRP gene promoter in 287 ostensibly healthy people. We identified two single-nucleotide polymorphisms (SNPs), a bi-allelic one at nucleotide -409 (G-->A), and a tri allelic one at -390 (C-->T-->A), both resident within the hexameric core of transcription factor binding E-box elements. Electrophoretic mobility shift assays confirmed that the SNP within the sequence (-412)CACGTG(-407) (E-box 1) modulates transcription factor binding, and that the one within (-394)CACTTG( 389) (E-box 2) supports transcription factor binding only when the -390 T allele is present. The commonest of four E-box 1/E-box 2 haplotypes (-409G/-390T) identified in the population supported highest promoter activity in luciferase reporter assays, and the rarest one (-409A/-390T) supported the least. Importantly, serum CRP in people with these haplotypes reproduced this rank order, i.e., people with the -409G/-390T haplotype had the highest baseline serum CRP (mean +/- SEM 10.9 +/- 2.25 microg/ml) and people with the -409A/-390T haplotype had the lowest (5.01 +/- 1.56 microg/ml). Furthermore, haplotype associated differences in baseline CRP were not due to differences in age, sex, or race, and were still apparent in people with no history of smoking. At least two other SNPs in the CRP promoter lie within E-box elements (-198 C-->T, E-box 4, and -861 T-->C, E-box 3), indicating that not only is the quality of E-box sites in CRP a major determinant of baseline CRP level, but also that the number of E-boxes may be important. These data confirm that the CRP promoter does encode functional polymorphisms, which should be considered when baseline CRP is being used as an indicator of clinical outcome. Ultimately, development of genetic tests to screen for CRP expression variants could allow categorization of healthy people into groups at high versus low future risk of inflammatory disease. PMID- 15778808 TI - Endothelial nitric oxide synthase gene polymorphism (Glu298Asp) in patients with coexistent hypertrophic cardiomyopathy and coronary spastic angina. AB - Coronary vasospasm appears to play a significant role in the etiology of myocardial ischemia in patients with hypertrophic cardiomyopathy (HCM). Furthermore, the management of patients with coexistent HCM and coronary spastic angina (CSA) presents a therapeutic challenge. The purpose of this study was to examine the Glu298Asp variant of the endothelial nitric oxide synthase (eNOS) gene to determine whether this polymorphism was associated with susceptibility to CSA in patients with HCM. The eNOS gene polymorphism (Glu298Asp) was genotyped in 150 HCM patients by the TaqMan chemical method. Patients were classified into group A (n=12) if they had CSA provoked by intracoronary acetylcholine, and group B (n=138) if they did not. In group A, the frequency of Glu/Glu, Glu/Asp, and Asp/Asp genotypes was 5 (41.7%), 6 (50%), and 1 (8.3%), respectively. In group B, it was 119 (86.2%), 17 (12.3%), and 2 (1.5%), respectively. The frequency of the Asp298 variant was significantly higher in group A than in group B (P<0.001). Multivariate logistic regression analysis showed that the Asp298 variant was a significant risk factor for CSA (odds ratio 11.8; P<0.001) that was independent of age, gender, smoking status or body mass index. Significantly more drugs were used by the patients in group A than those in group B and the patients with the Asp298 variant were treated with significantly more drugs than those without it. In conclusion, the Asp298 variant of the eNOS gene may be associated with CSA in HCM patients. HCM patients with CSA or the Asp298 variant may need more drugs to relieve their symptoms. PMID- 15778809 TI - Parkinsonism-hyperpyrexia syndrome not related to antiparkinsonian treatment withdrawal during the 2003 summer heat wave. PMID- 15778811 TI - Wegener granulomatosis presenting with haemorragic stroke in a young adult. PMID- 15778810 TI - Is lumbar spondylosis a cause of urinary retention in elderly women? AB - OBJECTIVES: Lumbar spondylosis (LS) is a common spinal degenerative disorder which causes various types of lower urinary tract dysfunction (LUTD). However, it is not certain whether LS may cause urinary retention in elderly women. METHODS: In a period covering the past 3 years, we retrospectively reviewed: a) urodynamic case records of women with urinary retention (post-void residuals, PVR > 100 ml), b) the records of women with LUTD due to LS (cauda equina syndrome and spinal canal narrowing by MRI), and c) uro-neurological features of women who belonged to both a) and b). RESULTS: a) One-hundred women with a mean age of 58 years had urinary retention. The most common underlying disease was multiple system atrophy [19], followed by multiple sclerosis [13] and cervical/thoracic tumours [8]. LS was the fourth most common [5], with the highest age (71 years) of all diseases. b) Nineteen women with LUTD had LS (12, canal narrowing of 50-70%; 7 > 70 %), with a mean PVR volume of 60 ml. A fourth [5] of them had urinary retention, with severe spinal canal narrowing (all 5 > 70%). c) Thus, 5 women belonged to both a) and b). In 4 of these women, LUTD followed or occurred together with typical cauda equina syndrome symptoms such as sciatica and saddle anesthesia. However, one elderly woman presented with painless urinary retention, and absent ankle reflexes were the sole neurological abnormality. The urodynamic abnormalities underlying urinary retention included an underactive detrusor in all 5, bladder sensory impairment in 3, an unrelaxing sphincter in 2, a low compliance detrusor in one, neurogenic sphincter motor unit potentials in 2 of 4 studied, and cholinergic supersensitivity of the detrusor in one of 3 studied. Surgical decompression ameliorated urinary retention in 1 of 2 women who had surgery. CONCLUSIONS: In our series, only 5 percent of the women with urinary retention had LS, but LS poses a potential risk for retention, particularly in elderly women with severe spinal canal narrowing. Preganglionic somato-autonomic dysfunctions underlie this condition. It may appear as the sole initial complaint in cases in which no other obvious neurological abnormalities are found. PMID- 15778812 TI - Acute disseminated encephalomyelitis associated with Campylobacter jejuni infection and antiganglioside GM1 IgG antibodies. PMID- 15778813 TI - Clinical characteristics of essential tremor in Mersin, Turkey--a population based door-to-door study. AB - OBJECTIVE: To describe the clinical features of essential tremor (ET) in a population. BACKGROUND: With few exceptions, clinical data on ET are derived from patients who attend specialty clinics. Most (> 90%) population-dwelling ET cases do not seek neurological attention. METHODS: 89 ET cases living in the Mersin province, Turkey were matched to 89 controls from the same population. All were examined by neurologists. Standardized scales included the Hamilton Depression Scale (HDS) and Hamilton Anxiety Scale (HAS). RESULTS: Eight-one (91%) of 89 cases previously had not been diagnosed as ET and 96.6% were untreated. Despite this,more than half (51.7%) of the cases answered "yes" to the question "are you disabled in some way by your tremor". Cases had more psychiatric symptomatology than controls (mean HDS scores = 11.4 +/- 8.2 vs. 7.9 +/- 6.1, p = 0.003 and mean HAS scores = 12.0 +/- 8.8 vs. 6.9 +/- 7.1, p < 0.001). Among ET cases, HDS scores (r = 0.24, p = 0.03) and HAS scores (r = 0.27, p = 0.01) were correlated with tremor severity. CONCLUSIONS: We present the clinical findings of a group of largely undiagnosed and untreated population-dwelling ET cases that would not otherwise have come to neurological attention. Approximately one-half reported functional difficulty and psychiatric symptoms were over-represented in these ET cases compared with matched controls. These findings suggest that ET, as it exists in the population, is not a completely benign entity. PMID- 15778814 TI - Interaction between interleukin-6 and intercellular adhesion molecule-1 genes and Alzheimer's disease risk. PMID- 15778815 TI - Crossed right hemisphere syndrome following left thalamic stroke. AB - In most right-handed people, language and motor functions are more reliant on systems of the left hemisphere while several non-linguistic visuo-spatial and attentional processes depend more on the right hemisphere. The rare exceptions to these rules provide important clues as to what functions co-lateralise, and are thus crucial for models of cerebral specialization. Here we report on the case of a patient, who, after a lesion restricted to the left thalamic region, showed signs normally associated with right hemispheric injury including motor impersistence, visuo-spatial dysfunction and poor comprehension of facial expression. Language abilities were spared and no signs of apraxia were present, in spite of his right hand, foot and eye preference, a pattern normally associated with conventional cerebral dominance. In spite of his other right hemispheric signs, the patient showed no signs of hemi-spatial neglect. The patient's pattern of spared and impaired abilities is compared and contrasted with other rare cases of crossed right hemisphere syndrome. PMID- 15778816 TI - Predicting the outcome of optic neuritis: evaluation of risk factors after 30 years of follow-up. AB - BACKGROUND: Multiple sclerosis (MS) is a common disease with considerable risk for disability. Optic neuritis (ON) is a common first symptom of MS but it can also remain an isolated episode. Therefore, predicting the outcome of ON has gained in importance, particularly in light of current discussions of early disease modifying treatments in individuals at risk of developing MS. We reported previously on our cohort of 86 patients with acute monosymptomatic unilateral ON of whom 33 had progressed to MS after up to 18 years. Three patients had died. The present study extends the observation period to 31 years. METHODS: Patients were followed for up to 31 years or until a diagnosis of MS was made. Cerebrospinal fluid (CSF) was examined at onset. HLA class I and II antigens were determined. Magnetic Resonance Imaging (MRI) was performed during follow up. FINDINGS: Only one of 50 patients at risk developed clinical manifestations of MS during the extended follow up period. The estimated 15-year-risk of MS was 40 % (confidence interval [CI] 31%-52%). Most cases, 20 of 34 or 60%, occurred within three years. Among factors present at onset, CSF with mononuclear pleocytosis and/or oligoclonal Ig increased the risk for subsequent MS significantly, 49% (CI 38%-65%) compared with 23 % (CI 12%-44%) for those with normal CSF, p=0.02. Younger patients and those with winter onset also had greater risk. Recurrence of ON similarly elevated the risk significantly, p<0.001. After 19-31 years MRI lesions suggestive of demyelinating disease were detected in 20 of 30 individuals although no clinical manifestations of MS had occurred. CONCLUSION: The risk of MS in this large population-based prospective ON patient series was 40% and significantly higher in those with inflammatory CSF abnormalities at onset. Clinically silent MRI lesions suggestive of MS were detected in a majority of those with "ON-only". This finding should be taken into account when discussing prognosis and early intervention in patients with clinically isolated ON. PMID- 15778817 TI - Permeability and reactivity of Thermotoga maritima in latex bimodal blend coatings at 80 degrees C: a model high temperature biocatalytic coating. AB - Thermostable polymers cast as thin, porous coatings or membranes may be useful for concentrating and stabilizing hyperthermophilic microorganisms as biocatalysts. Hydrogel matrices can be unstable above 65 degrees C. Therefore a 55-microm thick, two layer (cell coat + polymer top coat) bimodal, adhesive latex coating of partially coalesced polystyrene particles was investigated at 80 degrees C using Thermotoga maritima as a model hyperthermophile. Coating permeability (pore structure) was critical for maintaining T. maritima viability. The permeability of bimodal coatings generated from 0.8 v/v of a suspension of non-film-forming 800 nm polystyrene particles with high glass transition temperature (T(g) = 94 degrees C, 26.9% total solids) blended with 0.2 v/v of a suspension of film-forming 158 nm polyacrylate/styrene particles (T(g) approximately -5 degrees C, 40.9% total solids) with 0.3 g sucrose/g latex was measured in a KNO3 diffusion cell. Diffusivity ratio remained above 0.04 (D(eff)/D) when incubated at 80 degrees C in artificial seawater (ASW) for 5 days. KNO3 permeability was corroborated by cryogenic-SEM images of the pore structure. In contrast, the permeability of a mono-dispersed acrylate/vinyl acetate latex Rovace SF091 (T(g) approximately 10 degrees C) rapidly decreased and became impermeable after 2 days incubation in ASW at 80 degrees C. Thermotoga maritima were entrapped in these coatings at a cell density of 49 g cell wet weight/liter of coating volume, 25-fold higher than the density in liquid culture. Viable T. maritima were released from single-layer coatings at 80 degrees C but accurate measurement of the percentage of viable entrapped cells by plate counting was not successful. Metabolic activity could be measured in bilayer coatings by utilization of glucose and maltose, which was identical for latex-entrapped and suspended cells. Starch was hydrolyzed for 200 h by latex entrapped cells due to the slow diffusion of starch through the polymer top coat compared to only 24 h by suspended T. maritima. The observed reactivity and stability of these coatings was surprising since cryo-SEM images suggested that the smaller low T(g) polyacrylate/styrene particles preferentially bound to the T. maritima toga-sheath during coat formation. This model system may be useful for concentrating, entrapment and stabilization of metabolically active hyperthermophiles at 80 degrees C. PMID- 15778818 TI - Developmental morphology of the Asian one-leaf plant, Monophyllaea glabra (Gesneriaceae) with emphasis on inflorescence morphology. AB - We examined the developmental morphology of the tropical Asian one-leaf plant Monophyllaea glabra, which is believed to have diverged first in the phylogenetic tree of the genus. The embryo within the seed consists of two cotyledons and a hypocotyl with no shoot or root apical meristems. The endogenous root meristem is formed nearer the hypocotyl end than in other examined Monophyllaea species. One of the cotyledons grows to form the macrocotyledon by means of the basal meristem. The groove meristem arises between the anisocotyledons, shifts toward the macrocotyledon, and is transformed to the inflorescence apex, which produces inflorescence axes in the axils of all ventral bracts of two rows, and secondary inflorescences in the axils of the lower dorsal bracts of the other two rows. The macrocotyledon may act as a ventral bract for the first inflorescence axis at the reproductive stage. This organization suggests that a common ancestor of Monophyllaea and Whytockia with decussate inflorescences diverged in one direction to become Monophyllaea and in another to become Whytockia. PMID- 15778820 TI - Practical approach to the hormonal treatment of acne. AB - Acne is a disease of the pilosebaceous units and these are mainly under hormonal control. In female patients, hormonal therapy is a unique opportunity for the treatment of acne. Several combined oral contraceptives (COCs), cyproterone acetate, spironolactone, flutamide, and others, have been tried for the control of acne. An overview on the use of the most useful drugs in clinical practice was conducted. COCs are thoroughly discussed, also taking into consideration their potential side effects. A practical approach with guidelines on the use of COC in acne is proposed. PMID- 15778821 TI - Hormonal treatment of acne: review of current best evidence. AB - BACKGROUND: Recent controlled clinical trials have demonstrated the efficacy of various hormonal preparations, including oral contraceptives, in treatment of acne. OBJECTIVE: The goal of this article is to evaluate the best current evidence on the efficacy of hormonal treatment of acne. METHODS: English-language controlled clinical trials of systemic hormonal treatment of acne were sought by search of references of general dermatology texts, reviews on acne, drug product monographs, and computer-assisted search of MEDLINE, CINAHL, and COCHRANE databases between 1970 and 2003 using the key words hormonal, oral contraceptive, acne, spironolactone, cyproterone, flutamide, and therapy. For each agent, studies fulfilling the highest level of evidence were selected for further evaluation. RESULTS: The literature search recovered two placebo-controlled random controlled trials (RCTs) each of ethinyl estradiol 0.035 mg and norgestimate and ethinyl estradiol 20 microg and leonorgestrel 100 microg, three active-comparator RCTs of ethinyl estradiol 0.035 mg and cyproterone acetate 2 mg, one active comparator RCT of ethinyl estradiol 30 mug and drosperinone 3 mg, three small placebo-controlled RCTs of spironolactone, and one active-comparator RCT of flutamide. CONCLUSIONS: The efficacy of Tri-Cyclen and Alesse in acne is supported by high-quality RCTs (level A evidence). Evidence for the efficacy of Diane-35, spironolactone, and flutamide is derived from lower-quality RCTs (level B evidence). Results from a population-based epidemiological study suggests that Diane-35 is the most effective of these oral contraceptives in the treatment of acne (level B evidence; epidemiological study). PMID- 15778822 TI - Hormonal therapy of acne. PMID- 15778823 TI - Psychosocial effects of acne. AB - This article discusses the psychological effect of acne vulgaris. It is shown that acne has significant effect on self-image and impacts quality of life. The impact of acne may be equivalent to that of asthma or epilepsy. Anxiety and depression and a reduction in social functioning are a consequence of this condition. Effective treatment results in improvement of quality-of-life measurement. Most of the data is gathered from case control studies. Further work, particularly prospective longitudinal cohort studies, needs to be performed to validate the impact of acne on quality of life. Acne severity grading should incorporate life quality scores to better establish the true impact of this condition on our patients in order to optimize therapy. PMID- 15778824 TI - Hormonal approach to the treatment of acne a Canadian perspective. PMID- 15778825 TI - [Infection of the hip joint following psoas abscess. Case report and literature review]. AB - Infection of the hip joint following psoas abscess is uncommon. In fact, only two cases have been reported in the literature. The clinical aspect of psoas abscess is characterised only by unspecific back pain and hip pain, which are responsible for delayed diagnosis. MRT and culture of joint aspirate can permit early diagnosis. The preferred treatment is immediate surgical intervention with debridement, drainage and antibiotic therapy. Percutaneous drainage of psoas abscess alone only rarely is sufficient. In cases of infected hip joint, resection arthroplasty often cannot be avoided. Following complete decline of inflammatory parameters and certain absence of bacteria confirmed by repeated joint aspiration, prosthetic joint replacement is possible. This decision should be made on an individual basis, because Girdlestone hip also can lead to a satisfactory result concerning pain relief and functional outcome. PMID- 15778827 TI - [Avulsion fractures of the lower pole of the patella]. AB - Fractures of the lower pole of the patella are rare and occur typically in children. In these types of fractures an extensive sleeve of cartilage is pulled off with a small bony fragment. The so-called sleeve fractures are often overlooked in plain radiographs. We describe the operative treatment and outcomes in two patients with sleeve fractures, one receiving early and one delayed treatment.A 12-year-old boy (case 1) sustained an indirect injury to the left knee while playing ball. Clinical examination showed a lag of active extension of the left knee without decrease in passive range of motion. At 9 years of age, a 12-year-old girl (case 2) sustained a direct blunt trauma to her right knee while playing ball. The lesion of the lower pole of the patella was not diagnosed in time. During the following 3 years a lag of extension of the right knee developed.The sleeve fracture diagnosed early was treated by open reduction and internal fixation with transosseous suturing. At the 6-month follow-up the knee had regained full range of motion. The sleeve fracture diagnosed late showed a nonunion of the patella resulting in a lag of extension. Even after 3 years a shortening osteotomy of the patella resulted in full range of motion of the right knee. PMID- 15778826 TI - [Systemic capillary leak syndrome causes a compartment syndrome of both lower legs and a forearm. A case report with a review of the literature]. AB - Systemic capillary leak syndrome is a rare disorder which causes recurrent episodes of hypovolemic shock due to a markedly increased plasma shift into the interstitium. Hemoconcentration, hypoalbuminemia and a monoclonal gammopathy are characteristic laboratory findings. A rare manifestation of this disease is a compartment syndrome with rhabdomyolysis and acute renal failure. We describe a patient who suffers from a compartment syndrome of both lower legs and the left forearm secondary to systemic capillary leak syndrome, and discuss the pathophysiological background. PMID- 15778828 TI - [Fillet flaps as a possibility for defect reconstruction of the hand. Reconstruction without additional donor site morbidity]. AB - Fillet flaps offer an additional reconstruction opportunity for complex hand defects after trauma, burns, tumors or infections. This retrospective study elucidates the concept of fillet flaps and presents the results of an overall of 34 plastic surgical reconstructions of the hands in 31 patients. Pedicled axial pattern flaps were used predominantly, except 2 cross finger flaps. In 10 cases the defects were localized in the dorsal and in 9 cases in the palmar aspect of the hand. 14 finger defects and one of the ulnar hand were covered. Very few complications occured. In only 2 cases partial flap loss was observed. An additional wound infection required revision in one case. Another case was left to secondary healing. Prior to any amputation, possible use of spare parts for defect reconstruction should be considered as a matter of principle. Our data suggest that the concept of fillet flaps is suitable for the reconstruction of complex defects of the hands without additional donor site morbidity. PMID- 15778829 TI - [Value of arthroscopy in the treatment of upper ankle arthritis]. AB - OBJECTIVE: The purpose of this retrospective study was to evaluate arthroscopy in upper ankle osteoarthritis. METHODS: In the period from 1988 to 1997, 358 arthroscopies of the upper ankle joint were performed in 178 cases due to arthritis. Of 132 accessible patients, 124 replied to our questionnaire: 90 (73%) of the responders underwent clinical examination 94 (37-152) months following the initial arthroscopy and were graded according to the score of Evanski and Waugh. All arthroscopies were performed under fluid filling using anterolateral and anteromedial approaches. In only five cases was the posterior compartment additionally investigated. Generally, neither tourniquet nor mechanical joint distraction were applied. RESULTS: Of a total of 12 (6.7%) complications, only 3 (1.7%) hematomas needed surgical revision. One hematoma was due to an arteriovenous fistula treated by double ligation. All complications including five hypesthesias were temporary and subsided spontaneously. The 124 patients of the questionnaire group assessed the result of arthroscopic surgery as excellent in 11%, good in 46%, fair in 22%, and poor in 21%. Of all patients, 22% required further surgery of the upper ankle: in 9% further arthroscopy, in 3% arthrodesis, and in 10% microsurgical denervation. The total range of motion increased from 10/0/40 degrees (extension/flexion) preoperatively to 15/0/44 degrees at the follow-up examination. The Evanski score improved significantly (p<0.001) from 41 to 76 points. CONCLUSION: Due to minimal invasiveness and low risk of complications, arthroscopy is recommended for the following indications of upper ankle osteoarthritis: focal arthrosis, limited range of motion caused by osteophytes, soft tissue impingement, corpora libera, and synovitis. Severity and extent of upper ankle arthritis, range of motion, pain, local bone and soft tissue quality as well as the age, physical activity and compliance of the patient concerned are decisive for the individual therapeutic protocol. Alternative surgical techniques in upper ankle osteoarthritis are assessed such as denervation, distraction arthroplasty, correction osteotomy, ankle arthrodesis and total ankle replacement. PMID- 15778830 TI - [Force distribution in the wrist following scaphotrapeziotrapezoid arthrodesis]. AB - Scaphotrapeziotrapezoid (STT) arthrodesis has been proposed to stabilise the radial column and to redirect the load away from the lunate. Midterm effects on force transmission are still unclear. Six patients who were treated with triscaphe arthrodesis were examined after an average of 5 years using CT osteoabsorptiometry of both wrists. STT arthrodesis had been performed in four cases with Kienbock's disease type IIIb and in two cases of scaphotrapeziotrapezoid arthritis. At all contralateral wrists peak mineralisations were found beyond the lunate fossa and in the scaphoid fossa of the distal radius. At the side with STT arthrodesis there was only one peak. In five cases this density maximum was beyond the scaphoid fossa and in one case half beyond the lunate and half beyond the scaphoid fossa. Triscaphe arthrodesis allows load transmission from the lunate to the radial column. PMID- 15778831 TI - [Clinical results with A new retrograde femoral nail with a radiographical-free proximal locking device. A prospective study with 50 fractures]. AB - INTRODUCTION: Conventional retrograde nailing of the femur causes two important disadvantages: the proximal locking of the nail is difficult because of the anatomic conditions and a chondral defect was left into the knee. MATERIAL AND METHODS: After the retrograde implantation the new nail was lead through the greater trochanter. An additional proximal aiming device for proximal interlocking can be fixed. The entrance portal will be sealed by an osteochondral cylinder. 50 cases of femur fractures were selected for the prospective study. We recorded all intraoperative complications and technical difficulties. The cases were followed up for 52 weeks, both clinical and radiology examinations were performed. RESULTS: The mean follow up was 15.5+/-5 months. All fractures were healed. Knee movement was 125+/-14 degrees. The Leung Score was 84+/-12.6 points; HSS Score was 90+/-9 points. In two cases wound infections were developed. Mal union was observed in three cases, in two cases nail brake down. CONCLUSIONS: The new retrograde interlocking nail could be used to manage femur fractures successfully. Two aiming devices enable a easy interlocking. Replacement of the osteochondral cylinder into the entry portal reduces cartilage damage. PMID- 15778832 TI - [Surgical treatment of a displaced femoral head fracture with a cement-free dual headed prosthesis using a minimally invasive approach. Clinical and radiographic outcome]. AB - One hundred patients treated with a cementless bipolar prosthesis for a displaced subcapital hip fracture were prospectively evaluated for clinical and radiographic outcome. All patients were operated via a minimally invasive approach; in every case we implanted a Zweymuller stem. There were 77 women and 33 men with a mean age of 80 years (29-98 years). The mean duration of the operation was 29 min (20-95 min). Full weight bearing on crutches was allowed 1 day after the operation. The mean follow-up was 2.5 years (6 months to 7 years). Two years after the operation there were 40% of the remaining 65 patients who scored between 90 and 100 on the Harris hip score, 23% between 80 and 89, 20% between 70 and 79, and 17% below 70. No patient complained about thigh pain and up to now there has been no need for femoral revision due to loosening. There was no infection or nerve lesion. In three patients there was a luxation of the prosthesis which could be reduced by closed means. Radiographs from 81 patients showed stress shielding in 97.5% mainly in Gruen zones 1 and 7. Radiolucent lines in two or more Gruen zones were found in two patients. These findings suggest that the noncemented, pressfit, grit-blasted bipolar prosthesis demonstrated similar stability and radiographic results to cemented bipolar prostheses. Stress shielding was common but did not influence longevity of the implant. We did not find any signs of protrusion. Especially in older patients with a history of cardiac disease, the noncemented bipolar prosthesis is a rational alternative to avoid intra- and postoperative complications despite the higher costs for the implant. The minimally invasive approach helps to reduce operation time and intraoperative blood loss. PMID- 15778833 TI - [Epidural haematoma after operative evacuation of an acute subdural haematoma. Discussion of primary treatment illustrated by an unusual case]. AB - Subdural haematomas as a consequence of severe head injury often constitute the indication for operative evacuation. Despite intensive care management postoperative computed tomography scans are essential. This is illustrated by an unusual case report of a patient suffering from an epidural haematoma after operation of an subdural haematoma. In addition, the importance of the skull X ray in the emergency setting and intracranial pressure monitoring are discussed. PMID- 15778835 TI - Distal aneurysms of the unpaired ACA: embryologic and therapeutic aspects. AB - Anatomical variants of the cerebral arteries in general are frequent and due to the complex ontogenesis of these structures. Although encountered in many mammals, a single anterior cerebral artery (ACA) trunk is an infrequent finding in humans with an incidence of 3-5%. This vessel, giving rise to the arteries of both frontal lobes, is subjected to high flow volumes and distal arterial aneurysms have repetitively been encountered, mostly however before the introduction of endovascular treatment strategies. We report on five patients with acute SAH and arterial aneurysms of an unpaired ACA, who underwent coil embolisation. In all cases endovascular treatment using detachable platinum coils resulted in an at least satisfactory degree of aneurysm obliteration without parent artery occlusion or embolic infarcts. All patients had clinical and angiographic follow-up with median follow-up time of 29 months during which no aneurysm regrowth was encountered. In spite of a small patient group our results suggest, that altered flow dynamics due to enlarged single intracranial vessels may predispose to aneurysm formation and that endovascular embolisation is an appropriate treatment option in distal aneurysms of an unpaired ACA. PMID- 15778836 TI - Septic thrombosis of the transverse and sigmoid sinuses: imaging findings. AB - The purpose of this study was to evaluate the contribution of magnetic resonance imaging (MRI) to the diagnosis of septic thrombosis of transverse and sigmoid sinuses to analyze the different steps of the diagnosis and to identify the origin of the difficulties in diagnosis. This retrospective study included eight patients aged 53-81 years (mean age: 61.9 years) with proven or highly probable septic thrombosis of transverse and sigmoid sinuses. All patients underwent a pre and post-contrast enhancement brain CT scans and MRI. MR venogram (n=4) and HRCT of the temporal bone were performed when diagnosis was under discussion. After admission, the delay in diagnosis of lateral sinus thrombosis ranged from 8 to 60 days, with an average of 27 days (SD: +/-12.8). The delay in diagnosis was mainly due to non focused CT scans (6/8) or MR images performed at the initial presentation and absence of systematic radiological reading of the related fatty spaces and of skull base in bone windows (3/8). Diagnosis of septic origin of the thrombosis is of great importance, as it completely modifies the therapeutic planning of the patients. However, it remains a difficult challenge due to its lack of suggestive neurological or otolaryngologic symptoms. PMID- 15778837 TI - Renal artery entrapment by the diaphragmatic crus. AB - The aim of this study is to describe renal artery entrapment (RAE) by the diaphragmatic crus and to elucidate the diagnostic and therapeutic approach to this entity. From 1995 to 2002, 15 patients (mean age 65) were found to have a RAE. They were investigated by CT scan (n=14) and/or MRA (n=2) for hypertension (n=7), chronic renal insufficiency (n=4) or aneurysms (n=4). The right (n=11) or the left (n=4) renal artery (RA) was involved. The compression was ostial (n=8) or truncal (n=7), and was > or = 50% in eight cases. The course of the RA along the aorta on angiographic views (n=8) or a concentric ostial stenosis in a patient free of atheromatous lesions (n=7) were two findings suggestive of an RAE. Seven RAEs were indicated for treatment but only three were treated, by mean of stenting. Among the latter, two stents were patent at 6-month follow-up and one evolved to restenosis because of a stent fracture. RAEs may be suspected on angiographic views and proved by cross-sectional imaging because of specific imaging features. It is of importance to detect this etiology of RA stenosis because angioplasty with stenting is probably not always advisable. PMID- 15778838 TI - Prospective, multireader evaluation of image quality and vascular delineation of multislice CT angiography of the brain. AB - The aim of this prospective, multireader trial was to investigate image quality and vascular delineation of cranial multislice CT angiography (MSCTA) to identify strengths and weaknesses of the method. Sixty consecutive patients underwent standardized cranial MSCTA. The mean estimated effective dose was 0.96+/-0.11 mSv. Three masked readers independently graded image quality parameters and vascular delineation on a 5-point scale. Vascular attenuation values and dose length products were assessed quantitatively. Quantitative parameters were evaluated with a proportional odds regression model with bootstrapped standard errors to adjust the relevant standard errors for correlation within subjects and across readers. The non-parametric Wilcoxon sign-rank test was applied for quantitative measurements. Good to excellent ratings were observed regarding image quality parameters and vascular delineation. The delineation of veins was rated higher than that of arteries (OR 2.00). Smaller arterial segments were rated significantly less favorably than larger segments (OR up to 26.98). Moreover, the cavernous sinus, the C2 segment of the ICA and the communicating arteries demonstrated lower scores. Attenuation values were >240 HU and vessel-to parenchyma ratios >7 in all vessels. Cranial MSCTA achieved high ratings regarding image quality and vascular delineation. Relative weaknesses were found in small arterial subsegments and in vessels in close topographical proximity to bone. PMID- 15778839 TI - Pedal neuron 3 serves a significant role in effecting turning during crawling by the marine slug Tritonia diomedea (Bergh). AB - The marine nudibranch Tritonia diomedea crawls using its ciliated foot surface as the sole means of propulsion. Turning while crawling involves raising a small portion of the lateral foot margin on the side of the turn. The cilia in the lifted area no longer contribute to propulsion, and this asymmetry in thrust turns the animal towards the lifted side. Neurons located in the pedal ganglia of the brain contribute to these foot margin contractions. T. diomedea has a natural tendency to turn upstream (rheotaxis), and pedal flexion neuron Pedal 3 elicits foot margin lift and receives modulatory input from flow receptors. To assess the contribution of this single cell in turning behavior, two fine wires were glued to the surface of the brain over left and right Pedal 3. We determined that Pedal 3 activity is correlated with subsequent ipsilateral turns, preceding the lift of the foot margin and the change in orientation by a consistent interval. Both Pedal 3 cells show synchronous bursts of activity, and the firing frequency of the ipsilateral Pedal 3 increased before turns were observed to that side. Stimulation of the electrode over Pedal 3 proved sufficient to elicit an ipsilateral turn in Tritonia. PMID- 15778840 TI - Does an increase in reward affect the precision of the encoding of directional information in the honeybee waggle dance? AB - Apis mellifera foragers perform waggle dances to communicate the presence of highly desirable nectar sources to their forager-mates. Each waggle dance consists of several waggle-runs (straight movements of the dancer closely aligned on the comb surface) that carry spatial information that the dance followers can use to locate the food source being advertised. To address how this complex motor display responds to unpredictable fluctuations in its main triggering stimulus, i.e., sucrose stimulation, we analyzed the effects of an increase in reward on the direction of consecutive waggle-runs as well as other components of the waggle dance. Results show that a sudden increase in reward may increase the directional scatter among consecutive waggle-runs, especially those performed at the beginning of the dance. However, a simultaneous and rapid increase in the duration of the signal--together with a more regular alignment of the later waggle-runs within the signal--seems to compensate the initial increase in directional scatter so that the transfer of directional information remains effective. These results point out that the regulation of dance maneuvers depends on the dancer's motivation to forage. PMID- 15778842 TI - Macular hole and sub-hyaloid hemorrhage following filtering surgery with mitomycin C. AB - BACKGROUND: We report the occurrence of a macular hole with sub-hyaloid hemorrhage following filtering surgery with mitomycin C. METHODS: Combined trabeculotomy and trabeculectomy with mitomycin C was performed in the left eye for primary congenital glaucoma. RESULTS: Two weeks postoperatively the fundus showed a macular hole with subretinal and retinal hemorrhage along with sub hyaloid bleeding. Optical coherence tomography confirmed the presence of a full thickness macular hole. CONCLUSION: A full-thickness macular hole with subretinal and retinal hemorrhage is reported as a complication of glaucoma filtering surgery. PMID- 15778841 TI - Clinical and cytological findings in limbal stem cell deficiency. AB - PURPOSE: To evaluate and relate the clinical (including corneal sensitivity and tear function) and cytological (presence of goblet cells and cytokeratin 3- and 19-positive cells) features of limbal stem cell deficiency (LSCD). METHODS: Twenty-nine patients (44 eyes) with a clinical diagnosis of LSCD participated in this study. Corneal signs (epithelial alterations, superficial neovascularisation and stromal scarring) and cytological findings (presence of goblet cells and cytokeratins 3 and 19) were evaluated and scored (from 0 to 3) from each of the five corneal sectors. Corneal sensitivity (Cochet-Bonnet aesthesiometer) and tear function (Schirmer test and BUT) were also assessed. Cytological scores were correlated statistically with both corneal signs and sensitivity values. RESULTS: Cytokeratin 19-positive cells were found in 82% of corneal impression cytology samples, while goblet cells were identified in only 59% of these same samples. Cytokeratin 3-positive cells were present in 61% of LSCD eyes and in all unaffected eyes. Corneal sensitivity was significantly decreased in affected eyes compared with contralateral, healthy eyes (1.6+/-1.7 cm versus 5.7+/-0.3 cm). Tear function tests did not show significant changes. In LSCD eyes, goblet cells and cytokeratin 19-positive cells on the corneal surface were significantly correlated to corneal epithelial alterations and to corneal superficial neovascularisation (p<0.001). Corneal cytokeratin 3-positive cells were inversely related to epithelial alterations (p=0.003). Corneal sensitivity was decreased in corneal sectors with epithelial alterations (p<0.001), neovascularisation (p<0.001) and stromal abnormalities (p=0.049), and was indirectly related to the presence of goblet cells (p=0.005) and cytokeratin 19-positive cells (p<0.001). CONCLUSION: This study confirmed the importance of cytological tests in the diagnosis of LSCD. Furthermore, the absence of goblet cells may not exclude corneal conjunctivalisation as demonstrated by cytokeratin 19 immunostaining. Lastly, corneal conjunctivalisation was associated with zone-specific impairment in corneal sensitivity. PMID- 15778843 TI - Protein kinase C expression in the rabbit retina after laser photocoagulation. AB - BACKGROUND: Laser photocoagulation is a well-established treatment for diabetic retinopathy but the mechanism behind its effectiveness has not been elucidated. The protein kinase C (PKC) family is a group of enzymes which has been the subject of extensive interest in clinically related research since the advent of its role in the pathogenesis of diabetic retinopathy. With this study we wanted to explore whether PKC expression is altered in the retina after laser photocoagulation. METHODS: Normal rabbit eyes were treated with laser photocoagulation of varying intensity and examined after 30 min to 7 weeks. Treated and untreated regions of the retina were investigated histologically with the MC5 monoclonal antibody against PKC. Labeling for glial fibrillary acidic protein (GFAP), as well as hematoxylin and eosin (H&E) staining was also performed to assess the laser-induced trauma. RESULTS: In the normal retina, the MC5 antibody labeled rod bipolar cells and photoreceptor outer segments corresponding to PKC alpha. A translocated PKC expression with labeling concentrated in the rod bipolar terminals was seen in specimens examined 30 min after laser treatment, and after 1 week, no expression was seen in any part of the retina. After 2 weeks, PKC expression again indicated a translocated labeling pattern. After 5 weeks, labeling was found only in rod bipolar terminals in the peripheral retina. When comparing high- and low-intensity laser treatment 7 weeks postoperatively, no labeling was found in the high intensity-treated retinas, whereas low intensity-treated eyes displayed a near-normal labeling pattern. H&E staining revealed focal neuroretinal edema immediately after laser treatment, also in untreated areas. At later stages, destruction of the outer nuclear layer and migration of pigment epithelial cells in laser-lesioned areas was seen. GFAP labeled Muller cells were seen 1 week postoperatively in the entire retina. Labeling after this time decreased, but was still present in laser spots after 5 and 7 weeks. CONCLUSIONS: Laser photocoagulation alters the expression of PKC in the entire normal rabbit retina. The response follows a temporal pattern and is also related to laser intensity. These findings may help to explain the high efficacy of laser treatment in diabetic retinopathy. PMID- 15778844 TI - Inflammatory myofibroblastic tumor of the posterior mediastinum: an older adult case with anaplastic lymphoma kinase abnormalities determined using immunohistochemistry and fluorescence in situ hybridization. AB - Inflammatory myofibroblastic tumor (IMT) is a rare neoplasm that usually occurs in children and young adults. Anaplastic lymphoma kinase (ALK) abnormalities in IMT, determined using immunohistochemistry and/or molecular genetic studies, including fluorescence in situ hybridization (FISH), have almost been limited to children and young adults. In elderly cases of IMT, these ALK abnormalities are very rare. We report on a case of IMT arising in the posterior mediastinum of a 59-year-old Japanese man that showed ALK abnormalities determined using immunohistochemistry and FISH, suggesting the neoplastic nature of a subset of IMTs in older patients similar to those in younger ones and the presence of an additional mechanism(s) that allows them to start to grow late. PMID- 15778845 TI - Changes in the adherens junctions of human endothelial cells infected with spotted fever group rickettsiae. AB - Rickettsiae of the spotted fever group are obligately intracellular bacteria that primarily infect the vascular endothelium, invade adjacent cells propelled by actin polymerization, and cause severe systemic diseases. Endothelial dysfunction and vascular leakage develop as a consequence; this effect is the pathophysiological mechanism that explains most clinical manifestations. Here we report that rickettsial infection of cultured primary human endothelial cells is associated with the formation of gaps in the interendothelial adherens junctions, occurring late during the course of in vitro infections but not early, even when rickettsial loads are significant. PMID- 15778846 TI - Simplified quantification of urinary protein excretion using a novel dipstick in children. AB - Although the quantification of protein excretion is valuable for diagnosing and monitoring renal disease, accurate, timed, urine collection entails practical difficulties in children. Several authors have shown that the random urine protein/creatinine ratio (UP/UC) correlates well with timed protein excretion. A novel dipstick, Multistix PRO, has recently enabled us to analyze concentrations of both urinary protein and creatinine, semi-quantitatively, in 60 s. The aim of this study was to investigate whether the UP/UC values obtained by Multistix PRO correlate well with those obtained by quantitative methods and daily urinary protein excretion. In order to obtain the UP/UC values, we measured urinary protein and creatinine concentrations both semi-quantitatively by Multistix PRO and quantitatively by conventional methods. The relationship between the semi quantitative UP/UC by Multistix PRO and the quantitative UP/UC by conventional methods was analyzed. Similarly, the relationship between the semi-quantitative UP/UC and daily urinary protein excretion was studied. Semi-quantitative UP/UC by Multistix PRO correlated closely with both quantitative UP/UC and daily urinary protein excretion (r=0.86 and r=0.91, respectively). A cut-off level of heavy proteinuria, i.e., nephrotic range of proteinuria (>3.5 g/day) corresponded to 3.0, assessed by UP/UC by Multistix PRO. The semi-quantitative UP/UC by Multistix PRO correlated well with both quantitative UP/UC and daily urinary protein excretion, and use of the Multistix PRO would avoid errors and difficulties associated with timed urine collection. It is, therefore, a useful tool to monitor the urinary protein excretion in children with renal diseases at outpatient clinic. PMID- 15778847 TI - Pulmonary thrombosis in steroid-sensitive nephrotic syndrome. AB - Thrombo-embolic episodes are an uncommon but known complication of nephrotic syndrome. However, pulmonary thrombosis/thromboembolism is rare, especially in children. We describe the cases of two girls, aged 12 years, who presented with severe oedema in relapse. They had intermittent tachypnoea, and CT pulmonary angiography (CTPA) provided a less invasive and more definitive way of confirming pulmonary thrombosis/thromboembolism. They received heparin with resolution of the tachypnoea. Anticoagulation was continued for 6 months after the episode in one patient. They have been in remission for more than 1 year, and a thrombophilia screen does not indicate a predisposing tendency to the formation of clots. Pulmonary thrombosis/thromboembolism could present with subtle symptoms and needs prompt diagnosis and treatment to prevent a fatal outcome. PMID- 15778849 TI - Expression of the rat connexin 39 (rCx39) gene in myoblasts and myotubes in developing and regenerating skeletal muscles: an in situ hybridization study. AB - We report a detailed analysis of the expression pattern of the recently identified rat connexin gene, named rat connexin 39 (rCx39), both during embryonic development and in adult life. Qualitative and quantitative reverse transcription/polymerase chain reaction analysis showed intense expression of rCx39 restricted to differentiating skeletal muscles, with a peak of expression detected at 18 days of embryonic life, followed by a rapid decline to undetectable levels within the first week of postnatal life. A combination of the in situ hybridization technique for the detection of rCx39 mRNA and immunohistochemistry for myogenin, a myoblast-specific marker, allowed us to establish that the mRNA for this connexin was expressed in myogenin-positive myoblasts and early myotubes but disappeared in mature myotubes. Moreover, in adult animals, rCx39 mRNA was expressed in myogenic cells involved in skeletal myofiber regeneration following a crush injury. This is the first case of a connexin being mainly expressed in the myogenic cell lineage. The information presented should pave the way to novel molecular approaches in studies on the role of connexin-based gap-junctional communication in skeletal muscle differentiation and regeneration. PMID- 15778850 TI - Expression of lynx1 in developing lung and its modulation by prenatal nicotine exposure. AB - The expression of nicotinic acetylcholine receptors (nAChR) in fetal lung suggests maternal smoking during pregnancy effects newborn lung structure and function by the direct interaction of nicotine with nAChR in the developing lung. The recent identification of the lynx1 nAChR modulator protein in nicotinic neurons in the brain suggests that lynx1 may be similarly expressed in the lung. To study this, cDNAs encoding lynx1 were cloned from rhesus monkey lung. The temporal expression of lynx1 was studied in pre- and postnatal monkey lungs by in situ hybridization, immunohistochemistry, and realtime polymerase chain reaction (PCR). Lynx1 mRNA signal and lynx1 immunohistochemical staining were localized predominantly in airway epithelial cells, submucous glands, and smooth muscle cells, in endothelial and smooth muscle cells in vessel walls, and in alveolar type II cells. The distribution of lynx1 was similar to that of alpha4, beta2, and beta4 nAChR expression as determined by immunohistochemistry. Immunohistochemical staining also co-localized choline acetyltransferase, the enzyme that synthesizes acetylcholine, with lynx1 expression. Lynx1 expression was first observed in 71-day fetal lungs and increased with age. Immunohistochemistry, Western analysis, and realtime PCR analysis showed increased lynx1 expression in lungs following prenatal nicotine exposure. Thus, lynx1 is co-expressed with nAChR in the lung. Alteration of lynx1 levels is a potential new mechanism by which nicotine affects lung development. PMID- 15778852 TI - Effects of natural winter pasture and commercial pellet on the ultrastructure of small intestinal epithelium in reindeer. AB - Segments of small intestine (duodenum, jejunum and ileum) from slaughtered reindeer (Rangifer tarandus tarandus) grazing natural winter pastures (n=3) and reindeer fed commercially available pellets (RF-80) in winter (n=5) were collected and immediately fixed in McDowell's fixative. Transmission electron microscopy was employed to investigate the ultrastructural features of the epithelium and lamina propria along the small intestine and to relate these to the different diets. Major differences in ultrastructural features were observed between the small intestinal enterocytes of reindeer fed the two diets. Enterocytes in reindeer fed the natural diet displayed a normal appearance with a dense cytoplasm and distinct microvilli. In contrast, reindeer fed the commercial diet showed damaged enterocytes amongst the normal cells. Abnormal changes included disintegration and loss of microvilli, cytoplasmic swelling, loss of membrane integrity and increases in the width of intercellular spaces, especially in the jejunum. PMID- 15778851 TI - Comparison of effect of BMP-2, -4, and -6 on in vitro cartilage formation of human adult stem cells from bone marrow stroma. AB - The human adult stem cells from bone marrow stroma referred to as mesenchymal stem cells or marrow stromal cells (MSCs) are of interest because they are easily isolated and expanded and are capable of multipotential differentiation. Here, we examined the ability of recombinant human bone morphogenetic protein (BMP)-2, -4, and -6 to enhance in vitro cartilage formation of MSCs. Human MSCs were isolated from bone marrow taken from normal adult donors. The cells were pelleted and cultured for 21 days in chondrogenic medium containing transforming growth factor beta3 and dexamethasone with or without BMP-2, -4, or -6. All the BMPs tested increased chondrogenic differentiation as assayed by immunohistochemistry and by the size and weight of the cartilage synthesized. However, BMP-2 was the most effective. Microarray analyses of approximately 12,000 genes and reverse transcription-polymerase chain reaction assays established that the critical genes for cartilage synthesis were expressed in the expected time sequence in response to BMP-2. The tissue engineering of autologous cartilage derived from MSCs in vitro for transplantation will be a future alternative for patients with cartilage injuries. To obtain large amounts of cartilage rich in proteoglycans, the use of BMP-2 is recommended, instead of BMP-4 or -6. PMID- 15778853 TI - Localization of the Drosophila protein FL(2)D in somatic cells and female gonads. AB - The Drosophila gene female-lethal(2)d [fl(2)d] has been implicated in the alternative splicing regulation of genes involved in sexual determination, such as Sex-lethal and transformer, and in the alternative splicing of the gene Ultrabithorax. Here, the expression and subcellular localization of the FL(2)D protein is reported. FL(2)D is ubiquitously expressed in embryos, in imaginal discs of larvae and in ovaries. In Schneider cells, the protein is detected throughout the nucleoplasm, excluding the nucleolus, and partially co-localizes with SXL in nuclear speckles. In addition, FL(2)D associates with multiple transcriptionally active loci on salivary gland polytene chromosomes, consistent with the proposal that the protein is involved in the post-transcriptional regulation of a variety of genes. Interestingly, the localization of FL(2)D in nurse cells changes during oogenesis, from a rather diffuse early nuclear pattern in the germarium to a preferential accumulation at the nuclear periphery of nurse cells of developing cysts. PMID- 15778854 TI - Kappa opioid receptor is expressed in the rat cerebellar cortex. AB - Classically, the cerebellum has been shown to be involved in motor and visual functions, although recent evidence point to new roles of this organ. Pain processing is one of the recently described functions of the cerebellum. According to the importance of the opioid system in nociception, a detailed characterization of the expression pattern of opioid peptides in the cerebellum is the first step towards understanding the precise involvement of this organ in pain management. By using two different approaches (reverse transcription/polymerase chain reaction and in situ hybridization), we have detected, for the first time, expression of the kappa opioid receptor (KOR) gene in the cerebellar cortex of the rat. Expression is found in the molecular and granular layers in all the lobules of the cerebellum. Approximately 34% of the cells present in the molecular layer express KOR mRNA. This work contributes to the deeper knowledge of the mechanisms that are involved in cerebellar function and may lead to a better understanding of the relationships between nociceptive activity and drug abuse potential. PMID- 15778855 TI - Immunohistochemical evidence identifying the site of androgen production in the ovary of the protogynous grouper Epinephelus merra. AB - Androgen plays an important role in the developing ovaries of female fish. However, little is known regarding either the sites of production of androgen or its functional roles. In the present study, we investigated immunohistochemically the localization of cholesterol-side-chain-cleavage (P450scc) and cytochrome P45011beta-hydroxylase (P45011beta) with antibodies P450scc and P45011beta in the ovary of the female honeycomb grouper Epinephelus merra during its reproductive cycle. Clusters of strongly immunopositive cells, with 100-1000 cells in each cluster, against both P450scc and P45011beta, were observed throughout the annual reproductive cycle in tissue near blood vessels in the tunica ovary surrounding the outer periphery of the ovary. The ultrastructural characteristics of these cells showed that they were steroid-producing cells. In contrast, immunopositive cells against P450scc but not against P45011beta were localized in the theca layer surrounding the outer periphery of oocytes. These results suggest that two distinct steroid biosynthesis sites exist in the ovary and that cells at the two sites differ functionally. The only cells that biosynthesize 11-ketotestosterone are found in clusters in the vicinity of blood vessels; they possibly play a physiological role in oocyte growth and gonadal restructuring during the sex change of individuals of this species. PMID- 15778856 TI - Differentiation of epithelial cells in the urinary tract. AB - Uroplakins, cytokeratins and the apical plasma membrane were studied in the epithelia of mouse urinary tract. In the simple epithelium covering the inner medulla of the renal pelvis, no uroplakins or cytokeratin 20 were detected and cells had microvilli on their apical surface. The epithelium covering the inner band of the outer medulla became pseudostratified, with the upper layer consisting of large cells with stalks connecting them to the basal lamina. Uroplakins and cytokeratin 20 were not expressed in these cells. However, some superficial cells appeared without connections to the basal lamina; these cells expressed uroplakins Ia, Ib, II and III and cytokeratin 20, they contained sparse small uroplakin-positive cytoplasmic vesicles and their apical surface showed both microvilli and ridges. Cytokeratin 20 was seen as dots in the cytoplasm. This epithelium therefore showed partial urothelial differentiation. The epithelium covering the outer band of the outer medulla gradually changed from a two-layered to a three-layered urothelium with typical umbrella cells that contained all four uroplakins. Cytokeratin 20 was organized into a complex network. The epithelium possessed an asymmetric unit membrane at the apical cell surface and fusiform vesicles. Umbrella cells were also observed in the ureter and urinary bladder. In males and females, the urothelium ended in the bladder neck and was continued by a non-keratinized stratified epithelium in the urethra in which no urothelial cell differentiation markers were detected. We thus show here the expression, distribution and organization of specific proteins associated with the various cell types in the urinary tract epithelium. PMID- 15778857 TI - Comparative analysis of the expression patterns of Wnts during chick limb development. AB - Wnts control a number of processes during limb development-from initiating outgrowth and controlling patterning, to regulating cell differentiation in a number of tissues. We analyzed the expression pattern of various Wnts (4, 5a, 5b, 6, 11, and 14) in whole mount in situ hybridization during chick wing development. From HH stage 26, expression of Wnt 4 is observed in the central elbow region and wrist-forming regions, and during later stages, expression is seen in the joint-forming regions of the whole limb. Wnt 5a is expressed throughout the limb mesenchyme during early limb developmental stages, and later, at HH stage 23, it becomes predominantly confined to the distal tip, leaving low expression levels proximally. At HH stage 29, expression at the distal tip is restricted to the interdigital regions, and at day 8, expression is seen in the region surrounding the phalanges. Wnt 5b expression is first observed in the AER at HH stage 20 and later in the dorsal and ventral mesenchyme surrounding the cartilage elements of the limb. Expression of Wnt 6 is observed from HH stage 17 until day 8 in the dorsal and ventral ectoderm and also in the dorsoventral limb boundaries. Expression of Wnt 11 is observed in the proximal dorsal mesenchyme of the limb from HH stage 23 onward and later in the dorsal and ventral subectodermal mesenchyme and in the regions adjacent to the digits at day 8. Weak expression of Wnt 14 is observed at the proximal mesenchyme of the limb at HH stage 23; later, it extends as a transverse strip surrounding the cartilage elements as well as in the interdigital mesenchyme. PMID- 15778858 TI - Late-presenting congenital diaphragmatic hernia in children: a literature review. AB - This is a review of 122 articles published until December 2003 that describe 349 children with late-presenting postero-lateral congenital diaphragmatic hernia (CDH). Data regarding pre-operative diagnostic work-up were adequately reported in 177 patients with left CDH and in 41 with a right-sided defect. Chest radiography was the only diagnostic study in 92 (51.9%) children from the former group and in 17 (43.9%) from the latter group. In other patients, diagnostic work up encompassed various combinations of two or more imaging modalities. Apart from chest radiography, contrast study of the upper gastrointestinal tract was the most frequently performed imaging modality. In 88 (25.2%) children, initial radiographic features of CDH were misinterpreted. Pneumothorax and pleural effusion were the most common initial findings. Analysis of the hernial content in this group of patients has shown that herniation of the stomach, spleen or omentum should be regarded as risk factors for misdiagnosis of left CDH, whereas for right CDH, the risk factor is the presence of liver in the chest. Late presenting CDH may pose a significant diagnostic problem because of the great variability of radiographic appearance. Chest radiography following passage of a nasogastric tube and contrast studies of the gastrointestinal tract seem to be the most useful investigations for the diagnosis of left CDH. For patients with right CDH, owing to the high probability of liver herniation, a chest radiograph with liver scintigraphy or CT seems to be the best diagnostic option. PMID- 15778859 TI - The risk of venous thromboembolism is markedly elevated in patients with diabetes. AB - AIMS/HYPOTHESIS: Diabetes mellitus is associated with several changes in coagulation and fibrinolysis that may lead to a thrombogenic propensity. However, it is not known whether these perturbations actually cause increased risk of venous thromboembolism. METHODS: In a retrospective population-based study we evaluated the medical records of all 302 adult patients who were admitted to the Umea University Hospital with verified deep vein thrombosis or pulmonary embolism during the years 1997 to 1999. The patients were classified as diabetic (n=56) and non-diabetic (n=246) according to clinical information. The total number of diagnosed diabetic patients in different age groups in the catchment area was obtained from computerised registries in the primary health care centres and the Umea University Hospital, and data on the background population were collected from the Swedish population registry. RESULTS: The annual incidence rate of venous thromboembolism among diabetic patients in the population was 432 per 100,000 individuals (95% CI 375-496). In non-diabetic individuals it was 78 (95% CI 68-88). The age-adjusted incidence rate among the diabetic population was 274 (95% CI 262-286). The annual incidence rate of venous thromboembolism was elevated in type 1 and type 2 diabetic patients and the incidence rates were 704 (95% CI 314-1,566) and 412 (95% CI 312-544) respectively. The overall standardised morbidity ratio was 2.27 (95% CI 1.75-2.95), i.e. diabetic patients were more prone to venous thromboembolism after adjustment for age differences. CONCLUSIONS/INTERPRETATION: These results suggest that the age-adjusted risk for venous thromboembolism is more than two-fold higher among diabetic patients than in the non-diabetic background population. PMID- 15778860 TI - Adiponectin and beta cell dysfunction in gestational diabetes: pathophysiological implications. AB - AIMS/HYPOTHESIS: Gestational diabetes mellitus (GDM) identifies a population of young women at high risk of developing type 2 diabetes and thus provides an excellent model for studying early events in the natural history of this disease. Adiponectin, a novel adipocyte-derived protein with insulin-sensitising properties, has been proposed as a factor linking insulin resistance and beta cell dysfunction in the pathogenesis of type 2 diabetes. We conducted the current investigation to determine whether adiponectin is associated with beta cell dysfunction in GDM. METHODS: We studied 180 women undergoing OGTT in late pregnancy. Based on the OGTT results, participants were stratified into three groups: (1) NGT (n=93); (2) IGT (n=39); and (3) GDM (n=48). First-phase insulin secretion was determined using a validated index previously proposed by Stumvoll. Insulin sensitivity was assessed using the validated OGTT insulin sensitivity index of Matsuda and DeFronzo (IS(OGTT)). RESULTS: To evaluate beta cell function in relation to ambient insulin sensitivity, an insulin secretion-sensitivity index (ISSI) was derived from the product of the Stumvoll index and the IS(OGTT), based on the existence of the predicted hyperbolic relationship between these two measures. Mean ISSI was highest in the NGT group (6,731), followed by that in the IGT group (4,976) and then that in the GDM group (3,300) (overall p<0.0001), compatible with the notion of declining beta cell function across these glucose tolerance groups. Importantly, adiponectin was significantly correlated with ISSI (r=0.34, p<0.0001), with a stepwise increase in mean ISSI observed per tertile of adiponectin concentration (trend p<0.0001). In multivariate linear regression analysis, ISSI was positively correlated with adiponectin and negatively correlated with GDM, IGT and C-reactive protein (r(2)=0.54). CONCLUSIONS/INTERPRETATION: Adiponectin concentration is an independent correlate of beta cell function in late pregnancy. As such, adiponectin may play a key role in mediating insulin resistance and beta cell dysfunction in the pathogenesis of diabetes. PMID- 15778861 TI - Major differences in noradrenaline action on lipolysis and blood flow rates in skeletal muscle and adipose tissue in vivo. AB - AIMS/HYPOTHESIS: The regulation of skeletal muscle lipolysis is not fully understood. In the present study, the effects of systemic and local noradrenaline administration on lipolysis and blood flow rates in skeletal muscle and adipose tissue were studied in vivo. METHODS: First, circulating noradrenaline levels were raised tenfold by a continuous i.v. infusion (n=12). Glycerol levels (an index of lipolysis) were measured in m. gastrocnemius and in abdominal adipose tissue using microdialysis. Local blood flow was determined with the (133)Xe clearance technique and whole-body lipolysis rates assessed with a stable glycerol isotope technique ([(2)H(5)] glycerol). Second, interstitial glycerol levels in m. gastrocnemius, m. vastus and adipose tissue were measured by microdialysis during local perfusion with noradrenaline (10(-8)-10(-6) mol/l) (n=10). Local blood flow was monitored with the ethanol perfusion technique. RESULTS: With regard to systemic noradrenergic stimulation, no change in fractional release of glycerol (difference between tissue and arterial glycerol) was seen in skeletal muscle. In adipose tissue it transiently increased twofold (p<0.0001), and the rate of appearance of glycerol in plasma showed the same kinetic pattern. Blood flow was reduced by 40% in skeletal muscle (p<0.005) and increased by 50% in adipose tissue (p<0.05). After noradrenaline stimulation in situ, a discrete elevation of skeletal muscle glycerol was registered only at the highest concentration of noradrenaline (10(-6) mol/l) (p<0.05). Adipose tissue glycerol doubled already at the lowest concentration (10(-8) mol/l) (p<0.05). In skeletal muscle a decrease in blood flow was seen at the highest noradrenaline concentrations (p<0.05). CONCLUSIONS/INTERPRETATION: Lipolysis and blood flow rates are regulated differently in adipose tissue and skeletal muscle. Adipose tissue displays a high, but transient (tachyphylaxia) sensitivity to noradrenaline, leading to stimulation of both lipolysis and blood flow rates. In skeletal muscle, physiological concentrations of noradrenaline decrease blood flow but have no stimulatory effect on lipolysis rates. PMID- 15778862 TI - The demographic and behavioural profile of women with cervicitis infected with Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum and the comparison of two medical regimens. AB - OBJECTIVE: The aim of this study was to compare the therapeutic effect of single dose oral azithromycin with twice-daily, 7-day doxycycline in women with chlamydial, mycoplasmic or ureaplasmic cervicitis and to demonstrate the demographic and behavioral profile of infected women. MATERIALS AND METHODS: Five hundred and thirty-three women with various gynecologic complaints were recruited for this study. All women were screened for Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) by enzyme immune assay tests. Patients positive for Neisseria gonorrhoeae were excluded. Women treated for these infections were tested after completing medical therapy. Educational levels of infected women were similar in each group. The prevalence of CT, UU and MH was 3.4% (18/533), 11.8% (63/533) and 0.9% (5/533), respectively. In 452 patients, no treatment was administered. The remaining patients were either treated with azithromycin (n=41) or doxycycline (n=40). The eradication rate for the infectious agents was 87.3% and 93.5% in the group of azithromycin and doxycycline, respectively (P>0.05). There was no statistically significant difference in efficacy between single dose azithromycin and a 7-day course of doxycycline with respect to the treatment of culture-positive cases. Recurrences were observed in five cases in azithromycin group (12.5%) and in three cases in doxycycline group (7.5%). CONCLUSIONS: The treatment of uncomplicated chlamydial, mycoplasmic and ureaplasmic cervicitis with a single dose of azithromycin administered under supervision in the clinic is as effective as a 7-day course of doxycycline. This regimen may overcome the problem of compliance with the standard twice-daily, 7-day regimen of doxycycline. PMID- 15778864 TI - Microarray analysis of the Df1 mouse model of the 22q11 deletion syndrome. AB - The 22q11 deletion syndrome (22q11DS; DiGeorge/velo-cardio-facial syndrome) primarily affects the structures comprising the pharyngeal arches and pouches resulting in arch artery, cardiac, parathyroid, thymus, palatal and craniofacial defects. Tbx1 haploinsufficiency is thought to account for the main structural anomalies observed in the 22q11DS. The Df1 deleted mouse provides a model for 22q11DS, the deletion reflecting Tbx1 haploinsufficiency in the context of the deletion of 21 adjacent genes. We examined the expression of genes in Df1 embryos at embryonic day (E) 10.5, a stage when the arch-artery phenotype is fully penetrant. Our aims were threefold, with our primary aim to identify differentially regulated genes. Second, we asked whether any of the genes hemizygous in Df1 were dosage compensated to wild type levels, and third we investigated whether genes immediately adjacent to the deletion were dysregulated secondary to a position effect. Utilisation of oligonulceotide arrays allowed us to achieve our aims with 9 out of 12 Df1 deleted genes passing the stringent statistical filtering applied. Several genes involved in vasculogenesis and cardiogenesis were validated by real time quantitative PCR (RTQPCR), including Connexin 45, a gene required for normal vascular development, and Dnajb9 a gene implicated in microvascular differentiation. There was no evidence of any dosage compensation of deleted genes, suggesting this phenomenon is rare, and no dysregulation of genes mapping immediately adjacent to the deletion was detected. However Crkl, another gene implicated in the 22q11DS phenotype, was found to be downregulated by microarray and RTQPCR. PMID- 15778866 TI - Isolation and identification of bacteria from spent nuclear fuel pools. AB - The aim of the present research was to isolate and identify bacteria from spent nuclear fuel pools of a Spanish nuclear power plant. Water samples were collected and inoculated onto different culture media to isolate the highest number of species. 16S rDNA fragments from colonies growing on solid media were amplified and analysed by denaturing gradient gel electrophoresis. Sequencing revealed the presence of 21 different bacteria belonging to several phylogenetic groups (alpha, beta, and gamma-Proteobacteria, Actinomycetales, Flavobacterium, and the Bacillus/Staphylococcus group). The isolation of these microorganisms in this particular environment (oligotrophic and radioactive) is highly interesting because of the possibility of their being used for the bioremediation of radionuclide-contaminated waters. PMID- 15778867 TI - Cycling time trial performance during different phases of the menstrual cycle. AB - Submaximal exercise performance has not previously been assessed in the late follicular phase of the menstrual cycle, which is associated with a pre-ovulatory surge in oestrogen. Therefore, we compared cycling time trial performance during the early follicular (EF), late follicular (LF) and mid-luteal (ML) phase of the menstrual cycle in trained and untrained eumenorrhoeic women who cycled 30 and 15 km, respectively, in a non-fasted state. The women completed the three cycling time trials on a conventional racing bicycle mounted on an air-braked ergometer. We required resting oestrogen to increase by at least twofold above EF phase values in both the LF and ML phases and this resulted in a number of exclusions reducing the sample size of each group. No significant difference was noted in the finishing time between the different menstrual phases in trained (n=5) or untrained (n=8) group, albeit limited by sample size. However, analysis of the combined trained and untrained group data (n=13) revealed a trend for a faster finishing time (P=0.027) in the LF phase compared to the EF phase as 73% of the subjects showed improvements with an average of 5.2+/-2.9% (or 2.1+/-1.1 min) in the LF phase (for alpha=0.05 requires P<0.017). Combined group analysis yielded no difference between performance in the EF and ML phase or between the LF and ML phase. Thus, further research is encouraged to confirm the tendency for a faster time trial in the LF phase, which coincides with the pre-ovulatory surge in oestrogen. PMID- 15778869 TI - Positive reaction for cysticercosis and multicentric anaplastic oligoastrocytoma. AB - INTRODUCTION: An unusual case of positive immunological testing for cysticercosis in the cystic fluid obtained from an anaplastic oligoastrocytoma is presented. CASE REPORT: A 15-year-old boy was admitted with multiple brain lesions. The biggest was a cyst with a mural node and neurocysticercosis was suspected. In order to relieve intracranial pressure, the cyst was punctured and the immunological testing for cysticercosis was positive, reinforcing the clinical suspicion and leading to a clinical trial with albendazole and steroids. As the patient deteriorated the cystic lesion was removed and the diagnosis of anaplastic oligoastrocytoma was established. A second lesion was eventually approached and the histopathological diagnosis of both specimens concurred. DISCUSSION: Although some authors believe that chronic inflammatory changes following neurocysticercosis could induce the formation of brain tumors, this association may be a mere coincidence. In our case no clinical evidence of a prior infestation by Cysticercus was found. In fact, an exhaustive examination of the specimens did not reveal any areas of inflammatory reaction. We believe that the similarity of the glioma and cysticercosis antigens may be the cause of the positive reactions in the cystic fluid. PMID- 15778868 TI - Cross-species microarray hybridization to identify developmentally regulated genes in the filamentous fungus Sordaria macrospora. AB - The filamentous fungus Sordaria macrospora forms complex three-dimensional fruiting bodies that protect the developing ascospores and ensure their proper discharge. Several regulatory genes essential for fruiting body development were previously isolated by complementation of the sterile mutants pro1, pro11 and pro22. To establish the genetic relationships between these genes and to identify downstream targets, we have conducted cross-species microarray hybridizations using cDNA arrays derived from the closely related fungus Neurospora crassa and RNA probes prepared from wild-type S. macrospora and the three developmental mutants. Of the 1,420 genes which gave a signal with the probes from all the strains used, 172 (12%) were regulated differently in at least one of the three mutants compared to the wild type, and 17 (1.2%) were regulated differently in all three mutant strains. Microarray data were verified by Northern analysis or quantitative real time PCR. Among the genes that are up- or down-regulated in the mutant strains are genes encoding the pheromone precursors, enzymes involved in melanin biosynthesis and a lectin-like protein. Analysis of gene expression in double mutants revealed a complex network of interaction between the pro gene products. PMID- 15778870 TI - Intra-abdominal gout mimicking pelvic abscess. AB - Gout is the most common crystal-induced arthritis. Gouty tophi typically deposit in the extremities, especially toes and fingers. We present an unusual case of intrapelvic tophaceous gout in a patient suffering from chronic gouty arthritis. CT and MRI of the abdomen and pelvic cavity disclosed calcified gouty tophi around both hips, and a cystic lesion with peripheral enhancement in the pelvic cavity along the course of the iliopsoas muscle. The intra-abdominal tophus mimicked pelvic abscess. PMID- 15778871 TI - Single lamellar mechanics of the human lumbar anulus fibrosus. AB - The mechanical behavior of the entire anulus fibrosus is determined essentially by the tensile properties of its lamellae, their fiber orientations, and the regional variation of these quantities. Corresponding data are rare in the literature. The paper deals with an in vitro study of single lamellar anulus lamellae and aims to determine (i) their tensile response and regional variation, and (ii) the orientation of lamellar collagen fibers and their regional variation. Fresh human body-disc-body units (L1-L2, n=11) from cadavers were cut midsagittally producing two hemidisc units. One hemidisc was used for the preparation of single lamellar anulus specimens for tensile testing, while the other one was used for the investigation of the lamellar fiber orientation. Single lamellar anulus specimens with adjacent bone fragments were isolated from four anatomical regions: superficial and deep lamellae (3.9+/-0.21 mm, mean +/- SD, apart from the outer boundary surface of the anulus fibrosus) at ventro lateral and dorsal positions. The specimens underwent cyclic uniaxial tensile tests at three different strain rates in 0.15 mol/l NaCl solution at 37 degrees C, whereby the lamellar fiber direction was aligned with the load axis. For the characterization of the tensile behavior three moduli were calculated: E(low) (0 0.1 MPa), E(medium) (0.1-0.5 MPa) and E(high) (0.5-1 MPa). Additionally, specimens were tested with the load axis transverse to the fiber direction. From the second hemidisc fiber angles with respect to the horizontal plane were determined photogrammetrically from images taken at six circumferential positions from ventral to dorsal and at three depth levels. Tensile moduli along the fiber direction were in the range of 28-78 MPa (regional mean values). Superficial lamellae have larger E(medium) (p=0.017) and E(high) (p=0.012) than internal lamellae, and the mean value of superficial lamellae is about three times higher than that of deep lamellae. Tensile moduli of ventro-lateral lamellae do not differ significantly from the tensile moduli of dorsal lamellae, and E(low) is generally indifferent with respect to the anatomical region. Tensile moduli transverse to the fiber direction were about two orders of magnitude smaller (0.22+/-0.2 MPa, mean +/- SD, n=5). Tensile properties are not correlated significantly with donor age. Only small viscoelastic effects were observed. The regional variation of lamellar fiber angle phi is described appropriately by a regression line |phi|=23.2 + 0.130 x alpha (r(2)=0.55, p<0.001), where alpha is the polar angle associated with the circumferential position. The single anulus lamella may be seen as the elementary structural unit of the anulus fibrosus, and exhibits marked anisotropy and distinct regional variation of tensile properties and fiber angles. These features must be considered for appropriate physical and numerical modeling of the anulus fibrosus. PMID- 15778872 TI - Modelling of anisotropic growth in biological tissues. A new approach and computational aspects. AB - In this contribution, we develop a theoretical and computational framework for anisotropic growth phenomena. As a key idea of the proposed phenomenological approach, a fibre or rather structural tensor is introduced, which allows the description of transversely isotropic material behaviour. Based on this additional argument, anisotropic growth is modelled via appropriate evolution equations for the fibre while volumetric remodelling is realised by an evolution of the referential density. Both the strength of the fibre as well as the density follow Wolff-type laws. We however elaborate on two different approaches for the evolution of the fibre direction, namely an alignment with respect to strain or with respect to stress. One of the main benefits of the developed framework is therefore the opportunity to address the evolutions of the fibre strength and the fibre direction separately. It is then straightforward to set up appropriate integration algorithms such that the developed framework fits nicely into common, finite element schemes. Finally, several numerical examples underline the applicability of the proposed formulation. PMID- 15778873 TI - Brain temperature change and movement activation induced by intravenous cocaine delivered at various injection speeds in rats. AB - RATIONALE: Speed of intravenous (i.v.) injection presumably affects the rewarding effects of cocaine in humans. Work with animals has shown alterations in the behavioral and neurochemical effects of cocaine based on delivery speed. OBJECTIVES: We studied the effects of cocaine (1 mg/kg) as both a single i.v. injection and a series of five repeated injections (8-min intervals) delivered at different speeds (4, 16, and 64 s) on brain, muscle, and skin temperatures, and locomotion in awake, unrestrained rats. Since cocaine has a distinctive action on temperature, any enhancement of cocaine's properties by injection speed should readily be seen. RESULTS: When given as a single injection, cocaine increased brain temperature and locomotion, but transiently decreased skin and muscle temperatures; these effects were augmented by a high injection speed. Regardless of injection speed, however, changes in brain temperature and locomotion were strongly correlated with basal brain temperatures; higher temperatures were associated with less change after cocaine injection. When given as a series of five injections, cocaine increased brain temperature and locomotion. Although skin temperature initially decreased, it increased after successive cocaine injections. With each successive cocaine injection in the series, measures of temperature and movement parameters increased to a plateau and brain temperature change became biphasic. CONCLUSIONS: While confirming the results of others that rapid injection speed enhances cocaine-induced locomotor activation, our study suggests that delivery rate also affects the basic physiological actions of cocaine. PMID- 15778874 TI - Dronabinol and marijuana in HIV(+) marijuana smokers: acute effects on caloric intake and mood. AB - RATIONALE: No studies to date have directly compared the tolerability and efficacy of smoked marijuana and oral dronabinol in HIV(+) marijuana smokers. OBJECTIVES: The aim of this study was to compare dronabinol (0, 10, 20, 30 mg p.o.) and marijuana [0.0, 1.8, 2.8, 3.9% Delta(9)-tetrahydrocannabinol (THC)] in two samples of HIV(+) marijuana smokers: those with (n=15) and those without (n=15) a clinically significant loss of muscle mass (<90% body cell mass/height), which is one component of AIDS wasting. METHODS: Mood, physical symptoms, self selected food intake, cardiovascular data, and cognitive task performance were measured before and repeatedly after dronabinol and marijuana administration in eight 7-h sessions. Marijuana and dronabinol were administered in randomized order using a within-subject, staggered, double-dummy design. RESULTS: As compared to placebo, (1) marijuana (1.8, 2.8, 3.9% THC) and the lower dronabinol doses (10, 20 mg) were well tolerated (e.g., few physical symptoms, significant increases in ratings of "good drug effect") in both groups of participants; the highest dose of dronabinol (30 mg) was poorly tolerated in a subset of participants; (2) marijuana and dronabinol significantly increased caloric intake in the low bioelectrical impedance analysis (BIA) group but not in the normal BIA group; and (3) drug effects on cognitive performance were minor. CONCLUSIONS: These data suggest that for experienced marijuana smokers with clinically significant muscle mass loss, both dronabinol (at acute doses at least four to eight times the current recommendation) and marijuana produce substantial and comparable increases in food intake without producing adverse effects. PMID- 15778875 TI - Interaction of cocaine with positive GABAA modulators on the repeated acquisition and performance of response sequences in rats. AB - RATIONALE: Although positive GABA(A) modulators can attenuate several cocaine induced behavioral effects, there is a paucity of data on their interaction with cocaine on transition behavior or learning. OBJECTIVES: The current study examined the effects of cocaine (3.2-32 mg/kg), pregnanolone (3.2-24 mg/kg), and lorazepam (0.1-10 mg/kg) alone and in combination in rats responding under a multiple schedule of repeated acquisition and performance. METHODS: In the acquisition component, subjects acquired a different three-response sequence each session, whereas in the performance component, they responded on the same three response sequence each session. RESULTS: All three drugs produced dose-dependent rate-decreasing and error-increasing effects. Cocaine was the least effective in decreasing rates and the most effective in increasing the percentage of errors. In combination with pregnanolone (3.2 or 10 mg/kg), the rate-decreasing effects of cocaine were relatively unchanged in both components, but 3.2 mg/kg of pregnanolone enhanced its error-increasing effects and the 10-mg/kg dose produced a significant dose-dependent interaction on errors. The combination of cocaine with lorazepam (0.32 mg/kg, 70-min pretreatment) produced significantly greater rate-decreasing and error-increasing effects than cocaine alone. A 15-min pretreatment with the same dose of lorazepam enhanced the error-increasing effects of small doses and attenuated the effects of larger doses of cocaine. Combinations of pregnanolone and lorazepam produced greater rate-decreasing and error-increasing effects in both components than either drug alone. CONCLUSIONS: The present data show that cocaine is more disruptive to learning in rats than pregnanolone or lorazepam, and that the disruptive effects of cocaine can be enhanced by CNS depressants. PMID- 15778876 TI - 5-HT1A receptor expression during memory formation. AB - RATIONALE: It has been reported that 5-HT(1A) receptors modulate learning and memory and diverse pharmacological and genetic evidence supports this notion. Nevertheless, there are few works about expression of these receptors during memory formation. OBJECTIVE: We aimed to determine 5-HT(1A) receptor expression in brain areas of untrained, passive, and autoshaping trained groups of rats. METHODS: Ex vivo receptor autoradiography using the ligand agonist [(3)H]8 hydroxy-2-[di-n-propylamino]tetralin] (8-OH-DPAT) was used. RESULTS: The trained group relative to untrained animals showed increases of 5-HT(1A) receptor expression in 14 brain areas, decrements in 7, and no changes in 12. Thus, in contrast to untrained rats, 5-HT(1A) receptor expression of autoshaping trained rats was augmented in the tubercule olfactory, septal nucleus, nucleus accumbens, caudate putamen, globus pallidus, striate, and parietal (1 and 2), temporal cortex (1 and 3), granular retrosplenial cortex (1), amygdala, and median and dorsal raphe nuclei. In contrast, in the latter group, receptors were decreased in the CA1 area, hypothalamus dorsal, frontal cortex (1 and 3), occipital cortex, cingulate cortex (1 and 2), and cuneiform nucleus. There were significant differences between passive vs trained groups, but not regarding untrained rats, in the lateral olfactory tract, dentate gyrus, CA3 area, ventromedial hypothalamic, lateral hypothalamus, preoptic medial, frontal cortex (2), granular retrosplenial cortex (2), entorhinal cortex (1 and 2), piriform cortex, and substantia nigra. CONCLUSIONS: These data suggest that upregulated, downregulated, and "silence" of 5-HT(1A) receptors in brain areas form part of neural circuits engaged in memory formation by demonstrating a high degree of specificity and memory mapping. PMID- 15778877 TI - Involvement of the opioid system in the effects induced by nicotine on anxiety like behaviour in mice. AB - RATIONALE: Recent studies have revealed the participation of the endogenous opioid system in several behavioural responses induced by nicotine including antinociception, rewarding properties, and physical drug dependence. OBJECTIVES: The present study was designed to examine the possible involvement of the various opioid receptors in the anxiolytic- and anxiogenic-like responses induced by nicotine in mice. METHODS: The acute administration of low (0.05) or high (0.8 mg/kg) doses of nicotine subcutaneously produced opposite effects in the elevated plus maze, i.e. anxiolytic- and anxiogenic-like responses, respectively. Animals were only exposed once to nicotine. The effects of the pretreatment with the mu opioid receptor antagonist, beta-funaltrexamine (5 mg/kg), the delta-opioid antagonist, naltrindole (2.5 mg/kg) and the kappa-opioid antagonist, nor binaltorphimine (2.5 mg/kg) intraperitoneally were evaluated on the anxiolytic- and anxiogenic-like responses induced by nicotine. RESULTS: beta-funaltrexamine, but not nor-binaltorphimine or naltrindole, abolished nicotine-induced anxiolytic like effects, suggesting an involvement of mu-opioid receptors in this behavioural response. On the other hand, naltrindole, but not nor-binaltorphimine or beta-funaltrexamine, increased the anxiogenic-like responses of nicotine, suggesting an involvement of delta-receptors in this behavioural effect. CONCLUSIONS: These results demonstrate that the endogenous opioid system is involved in the effects induced by nicotine on anxiety-like behaviour and provide new findings to further clarify the interaction between these two neurochemical systems. PMID- 15778878 TI - Somatostatin receptors in the ventral pallidum/substantia innominata modulate rat locomotor activity. AB - RATIONALE: Somatostatin and its receptors (sst(1) and sst(2)) have been localized in brain nuclei implicated in motor control, such as the nucleus accumbens, ventral pallidum (VP) and substantia innominata (SI). OBJECTIVES: The objective of the study is to investigate the effect of somatostatin and selective sst(1) and sst(2) analogs infused in the VP/SI on the locomotor activity of the rat. METHODS: Somatostatin (15, 30, 60, 120 and 240 ng/0.5 microl/side), CH275 (sst(1) analog; 60, 180, 240 and 480 ng/0.5 microl/side), MK678 (sst(2) analog; 120, 240 and 480 ng/0.5 microl/side), L-809,087 (sst(4) agonist, 240 ng/0.5 microl/side) or saline (vehicle) were infused bilaterally in the VP/SI of the rat and locomotor activity measured for 60 min. The effect of SRA-880 (sst(1) antagonist) and CYN-154806 (sst(2) antagonist) on somatostatin-, CH275- and MK678-mediated locomotor activity was also ascertained. RESULTS: Somatostatin decreased locomotor activity in the first 30 min after its infusion in the VP/SI and in a dose-dependent manner. The sst(1) and sst(2) antagonists, SRA-880 and CYN-154806, respectively, reversed the somatostatin effect. The sst(1) and sst(2) agonists CH275 and MK678, respectively, mimicked somatostatin's actions, while the selective sst(4) agonist L-809,087 had no effect. Moreover, SRA-880 and CYN 154806 reversed the respective agonist action on locomotor activity. CONCLUSION: The present study provides functional evidence for the presence of sst(1) and sst(2) receptors in the VP/SI and their implication in motor control. The mechanism via which somatostatin and agonists mediate the attenuation of locomotor activity is presently being investigated. PMID- 15778879 TI - Sleep-related vagotonic effect of zolpidem in rats. AB - RATIONALE: Zolpidem is a relatively new nonbenzodiazepine sedative-hypnotic. The effects of zolpidem on autonomic functions remain unclear. OBJECTIVES: The aim of this study was to evaluate the effects of zolpidem on sleep and related cardiac autonomic modulations as compared with triazolam in Wistar-Kyoto rats. METHODS: Continuous power spectral analyses of electroencephalogram (EEG), electromyogram, and heart rate variability were performed on freely moving rats during daytime sleep. The consciousness states were classified into active waking (AW), quiet sleep (QS), and paradoxical sleep (PS). Drugs were administered via gavage and data within 2 h were analyzed. RESULTS: All zolpidem (ZP3, 3 mg/kg; ZP30, 30 mg/kg) and triazolam (TZ0.075, 0.075 mg/kg; TZ0.75, 0.75 mg/kg) groups had longer accumulated QS time and averaged QS duration as compared with the vehicle control. The accumulated QS time and averaged QS duration of ZP3 were similar to those of TZ0.075. Significant suppressions of PS time were noted in all drug groups except ZP3. During QS, ZP3 and ZP30 exhibited significant increases of magnitude and percentage of EEG delta power, whereas TZ0.075 and TZ0.75 did not. Heart period and high-frequency power of heart rate variability increased significantly in ZP3 during all sleep-wake states. Both parameters, however, did not increase but even decreased in ZP30, TZ0.075, and TZ0.75. CONCLUSIONS: Zolpidem not only caused a longer and deeper sleep but also led to an elevated cardiac vagal activity at a specific dose in the rat. PMID- 15778880 TI - Added donepezil for stable schizophrenia: a double-blind, placebo-controlled trial. AB - RATIONALE: Schizophrenia is a disorder with cognitive deficits that could stem from cholinergic dysfunction. OBJECTIVES: Our aim was to examine if donepezil administered to stable, medicated outpatients with schizophrenia improves cognition and psychopathology. METHODS: We conducted a double-blind placebo controlled trial of donepezil up to 10 mg/day added for 8 weeks to ongoing antipsychotic treatment in 36 typical community-treated schizophrenia patients not selected for cognitive impairment. RESULTS: Donepezil did not improve measures of cognition or psychopathology. It was well tolerated. CONCLUSION: Consistent with other studies, addition of donepezil to stable patients with schizophrenia did not improve cognition or measures of psychopathology. This result does not support the hypothesis that residual symptoms and cognitive problems result from a cholinergic deficit that can be remedied by an acetylcholinesterase inhibitor. A donepezil add-on strategy might make sense in selected schizophrenia cases where a pathological process is known to affect cholinergic neurons (e.g., history of head injury or comorbid dementia). PMID- 15778881 TI - Acetylcholinesterase inhibitors ameliorate behavioral deficits in the Tg2576 mouse model of Alzheimer's disease. AB - RATIONALE: Acetylcholinesterase inhibitors are widely used for the treatment of patients with Alzheimer's disease (AD). However, the relationship between the capacity of such drugs to ameliorate the symptoms of AD and their ability to alter the underlying disease process is not well understood. Transgenic mice that overexpress the human form of amyloid precursor protein and develop deposits of beta-amyloid (Abeta) and behavioral deficits during adulthood are useful for investigating this question. OBJECTIVES: The effects of administration of two acetylcholinesterase inhibitors, physostigmine and donepezil, on Abeta plaque formation and memory-related behaviors were investigated in the Tg2576-transgenic mouse model of AD. At 9-10 months of age, Tg2576-transgenic [Tg(+)] mice develop Abeta plaques and impairments on paradigms related to learning and memory as compared to transgene-negative [Tg(-)] mice. METHODS: Beginning at 9 months of age, increasing doses of physostigmine (0.03, 0.1, and 0.3 mg/kg), donepezil (0.1, 0.3, and 1.0 mg/kg), or saline were administered over 6 weeks to cohorts of Tg(+) and Tg(-) mice. Performance on tests of spatial reversal learning and fear conditioning was evaluated at each drug dose throughout the period of drug administration. After drug administration was completed, the animals were sacrificed and Abeta plaque number was quantified. RESULTS: Administration of physostigmine and donepezil improved deficits in contextual and cued memory in Tg(+) mice so that their behaviors became more similar to Tg(-) mice. However, administration of physostigmine and donepezil tended to improve cued memory and deficits in spatial learning in both Tg(+) and Tg(-) mice. Physostigmine administration demonstrated more prominent effects in improving contextual memory than donepezil, while donepezil was more effective than physostigmine in improving deficits in the acquisition of the spatial memory paradigm. Administration of neither drug altered the deposition of Abeta plaques. CONCLUSIONS: These studies suggest that acetylcholinesterase inhibitors can ameliorate memory deficits in Tg(+) mice without necessarily altering the deposition of Abeta plaques. Tg2576 mice may be useful as an animal model to further investigate the mechanisms by which aceytlcholinesterase inhibitors improve cognitive deficits in patients with AD. PMID- 15778883 TI - Effects of alcohol and performance incentives on immediate working memory. AB - RATIONALE AND OBJECTIVES: The Sternberg Memory Scanning task was used to test the hypotheses that immediate working memory (WM) would be impaired during moderate rising blood alcohol concentrations (BACs), and provision of a rewarding incentive for good performance under alcohol would counteract this impairment. METHODS: Two groups (n=18) of social drinkers received either 0.62 g/kg alcohol (A) or a placebo (P). Reward (verbal feedback and money) for good performance was provided to another pair of alcohol and placebo groups (n=18) (AR and PR). All participants completed two tests when BACs in the alcohol groups were rising (68 and 80 mg/100 ml) and two tests during declining BACs (73 and 64 mg/100 ml). RESULTS: Comparisons of A and P groups showed that rising BACs impaired immediate WM [increased errors, slowed rate of mental scanning, slowed reaction time (RT) when a larger number of items had to be held in WM]. Comparisons of AR and PR groups showed that reward for good performance during rising BACs counteracted impairment in scanning rate and RT but did not reduce errors. Tests during declining BACs of 73 mg/100 ml indicated some symptoms of impairment abated before others. When BAC declined to 64 mg/106 ml, the groups no longer differed on any measure of WM. CONCLUSIONS: An acute dose of alcohol can impair aspects of immediate WM performance while BACs are rising, and some of these effects can be reversed when incentives to perform well are provided. PMID- 15778882 TI - Screening antidepressants in the chick separation-stress paradigm. AB - RATIONALE: Clinical research has indicated that antidepressants are efficacious in the treatment of anxiety disorders, especially when repeatedly administered. However, few animal models of anxiety are sensitive to antidepressants, a finding that may be due to procedures limited to acute administrations. OBJECTIVES: The purpose of the present research was to further validate the chick separation stress paradigm as an animal model of anxiety by examining its sensitivity to the monoamine oxidase inhibitor (MAOI) phenelzine (6.25, 12.5, 25.0 mg/kg), the tricyclic antidepressant (TCA) imipramine (5.0, 10.0, 20.0 mg/kg), the selective serotonin reuptake inhibitor (SSRI) citalopram (1.0, 2.5, 5.0 mg/kg), and the norepinephrine reuptake inhibitor (NRI) maprotiline (5.0, 10.0, 20.0 mg/kg) under acute (no pretreatment) or repeated (3 or 6 days pretreatment) administration procedures. METHODS: Following any pretreatment, 8-day-old chicks received their respective vehicle or drug probe injection 15 min before tests in either a "mirror" (low stress) or "no mirror" (high stress) condition for a 180-s isolation period. The dependent measures were distress vocalizations to index separation stress and sleep onset latency to index sedation. RESULTS: The model was sensitive to acutely administered phenelzine (MAOI), imipramine (TCA), and maprotiline (NRI), but not citalopram (SSRI) and retained its sensitivity to these drug probes across both repeated administration procedures. None of the drug probes possessed any sedative properties. CONCLUSIONS: These results help extend the validity and utility of the chick separation-stress paradigm as an animal model of anxiety by demonstrating its sensitivity to antidepressants under both acute and repeated administration procedures. PMID- 15778884 TI - A parametric analysis of olanzapine-induced weight gain in female rats. AB - RATIONALE: Some novel antipsychotics, including olanzapine, induce weight gain and metabolic abnormalities, which represent the major adverse effects of these drugs. However, the mechanism(s) involved in such effects are unclear. OBJECTIVE: The aim of this study was to develop, in female rats, a parametric model of olanzapine-induced weight gain and metabolic abnormalities and evaluate it against clinical findings. METHODS: Female rats were administered olanzapine b.i.d. at doses of 0, 1, 2 and 4 mg/kg over 20 days, and a wide range of variables were recorded during and after drug administration. RESULTS: Olanzapine increased both 24 h and total food intake. This was associated with rapid onset weight gain and increased adiposity (assessed by visceral fat pad masses). Insulin, but not glucose, concentrations were elevated, with a significant increase in the HOMA-IR index, indicative of insulin resistance. A nonsignificant trend towards higher levels of leptin was observed. Paradoxically, there was a significant increase in adiponectin. All of these variables showed maximal increases at either 1 or 2 mg/kg and attenuated effects at 4 mg/kg. Prolactin levels were also increased by olanzapine. However, for this variable, there was a clear dose-response curve, with the maximal effect at the highest dose (4 mg/kg). CONCLUSIONS: These data suggest that aspects of olanzapine-induced weight gain and metabolic abnormalities can possibly be modelled in female rats. It is suggested that olanzapine-induced hyperphagia acts as an initial stimulus which leads to weight gain, enhanced visceral adiposity and subsequent insulin resistance, although the latter may be ameliorated by compensatory responses in adiponectin levels. Prolactin elevation appears likely not to be involved in the weight gain, adiposity and metabolic changes seen in this model. PMID- 15778885 TI - Repeated administration of methylphenidate in young, adolescent, and mature rats affects the response to cocaine later in adulthood. AB - RATIONALE: Previous studies have shown that the expression of behavioral sensitization to psychostimulants depends on the age and gender of the animal. OBJECTIVE: This study was conducted to determine the pattern of behavioral sensitization to repeated administration of methylphenidate (MPD) at three different developmental ages and to assess the response to a cocaine challenge in adulthood. METHODS: We gave five daily i.p. injections of 10 or 20 mg kg(-1) of MPD (10 MPD, 20 MPD) or saline to male and female rats beginning on postnatal days (PND) 21, 45, or 60. When all groups reached PND 90, rats were challenged with 10 mg kg(-1) cocaine. For both MPD administration and cocaine challenge, locomotion and stereotyped behaviors were assessed for 1 h. RESULTS: The 10 MPD dose produced increased locomotion over the other two treatments at all ages. Rats that received 20 MPD showed a decline in locomotion across days with an increase in the time spent in high intensity stereotypy by day 5. Animals treated with 10 MPD showed diverse behavioral responses with adolescents showing somewhat dampened stereotypy than the other two age groups. In response to cocaine, pretreatment with MPD at all ages enhanced the cocaine response and produced qualitatively different patterns of stereotyped behavior for each gender and pretreatment age group. CONCLUSION: MPD produced clear age-specific sensitization of behavior in rats. Furthermore, exposure to MPD cross-sensitized with cocaine regardless of the age at which MPD exposure occurred with each pretreatment age group showing a unique pattern of responses. PMID- 15778886 TI - Acute effects of nicotine on visual search tasks in young adult smokers. AB - RATIONALE: Nicotine is known to improve performance on tests involving sustained attention and recent research suggests that nicotine may also improve performance on tests involving the strategic allocation of attention and working memory. OBJECTIVES: We used measures of accuracy and response latency combined with eye tracking techniques to examine the effects of nicotine on visual search tasks. METHODS: In experiment 1 smokers and non-smokers performed pop-out and serial search tasks. In experiment 2, we used a within-subject design and a more demanding search task for multiple targets. In both studies, 2-h abstinent smokers were asked to smoke one of their own cigarettes between baseline and tests. RESULTS: In experiment 1, pop-out search times were faster after nicotine, without a loss in accuracy. Similar effects were observed for serial searches, but these were significant only at a trend level. In experiment 2, nicotine facilitated a strategic change in eye movements resulting in a higher proportion of fixations on target letters. If the cigarette was smoked on the first trial (when the task was novel), nicotine additionally reduced the total number of fixations and re-fixations on all letters in the display. CONCLUSIONS: Nicotine improves visual search performance by speeding up search time and enabling a better focus of attention on task relevant items. This appears to reflect more efficient inhibition of eye movements towards task irrelevant stimuli, and better active maintenance of task goals. When the task is novel, and therefore more difficult, nicotine lessens the need to re-fixate previously seen letters, suggesting an improvement in working memory. PMID- 15778887 TI - Individual differences in initial sensitivity and acute tolerance predict patterns of chronic drug tolerance to nitrous-oxide-induced hypothermia in rats. AB - RATIONALE: A preventive strategy for drug addiction would benefit from being able to identify vulnerable individuals. Understanding how an individual responds during an initial drug exposure may be useful for predicting how that individual will respond to repeated drug administrations. OBJECTIVES: This study investigated whether individual differences in initial drug sensitivity and acute tolerance can predict how chronic tolerance develops. METHODS: During an initial 3-h administration of 60% nitrous oxide (N(2)O), male Long-Evans rats were screened for N(2)O's hypothermic effect into subsets based on being initially insensitive (II), sensitive with acute tolerance (AT), or sensitive with no intrasessional recovery (NR). Animals in each individual difference category were randomly assigned to receive six 90-min exposures of either 60% N(2)O or placebo gas. Core temperature was measured telemetrically. RESULTS: Rats that exhibited a comparable degree of hypothermia during an initial N(2)O exposure, but differed in acute tolerance development, developed different patterns of chronic tolerance. Specifically, the NR group did not become fully tolerant over repeated N(2)O exposures while the AT group developed an initial hyperthermia followed by a return of core temperature to control levels indicative of full tolerance development. By the second N(2)O exposure, the II group breathing N(2)O became hyperthermic relative to the placebo control group and this hyperthermia persisted throughout the multiple N(2)O exposures. CONCLUSIONS: Individual differences in initial drug sensitivity and acute tolerance development predict different patterns of chronic tolerance. The hypothesis is suggested that individual differences in opponent-adaptive responses may mediate this relationship. PMID- 15778888 TI - Social and environmental influences on opioid sensitivity in rats: importance of an opioid's relative efficacy at the mu-receptor. AB - RATIONALE: Evidence indicates that social and environmental enrichment can influence the functional maturation of the central nervous system and may affect an organism's sensitivity to centrally acting drugs. OBJECTIVE: The purpose of the present study was to examine the effects of social and environmental enrichment on sensitivity to mu-opioids possessing a range of relative efficacies at the mu-receptor. METHODS: Rats were obtained at weaning (21 days) and divided into two groups immediately upon arrival. Isolated rats were housed individually in opaque laboratory cages with no visual or tactile contact with other rats; enriched rats were housed socially in groups of four in large cages and given various novel objects on a daily basis. After 6 weeks under these conditions, the effects of morphine, levorphanol, buprenorphine, butorphanol, and nalbuphine were examined in the warm-water, tail-withdrawal procedure and the place-conditioning procedure. RESULTS: In the tail-withdrawal procedure, isolated and enriched rats did not differ in sensitivity to morphine (1.0-30 mg/kg) and levorphanol (0.3-10 mg/kg), but enriched rats were more sensitive to buprenorphine (0.03-3.0 mg/kg), butorphanol (0.3-30 mg/kg), and nalbuphine (0.3-30 mg/kg). In drug combination tests, butorphanol and nalbuphine antagonized the effects of morphine in isolated rats under conditions in which they produced high levels of antinociception in enriched rats. In the place-conditioning procedure, doses of 10 morphine and 3.0 levorphanol established a place preference in both groups of rats, whereas doses of 0.3 buprenorphine, 3.0 butorphanol, and 10 nalbuphine established a place preference only in enriched rats. CONCLUSIONS: These findings may be taken as evidence that enriched rats are more sensitive than isolated rats to the effects of lower-efficacy mu-opioids and that social and environmental enrichment leads to functional alterations in opioid receptor populations. PMID- 15778892 TI - Repeated cold showers as a method of habituating humans to the initial responses to cold water immersion. AB - The hypothesis that the initial responses to cold water immersion could be attenuated by repeated cold showers was tested. Eighteen (13 men, 5 women) non habituated subjects undertook two 3-min head-out seated immersions into stirred water at 10 degrees C wearing swim wear. The immersions were separated by 4 days during which time they took six cold showers. The subjects were randomly split into three groups with different showering regimes: 3 min at 10 degrees C on the back (10B); 3 min at 15 degrees C on the back (15B); and 30 s at 10 degrees C on the back followed by 30 s on the front (10BF). Over the first 30 s of immersion respiratory frequency ( f (R)) was reduced by 21% in groups 10B and 10BF from 54 (14) to 44 (16) breaths.min(-1) ( P <0.05), and 33 (8) to 26 (10) breaths.min(-1) ( P <0.05) respectively, following repeated showers. Group 15B showed no change in f (R). The tachycardia induced on immersion in water at 10 degrees C was not reduced by repeated showers except in group 15B during the last 150 s [from 119 (23) to 105 (25) beats.min(-1), P <0.05]. Repeated showering in water at 10 degrees C reduced the respiratory drive (as measured by f (R)) during head-out immersion in water at the same temperature. No such habituation was observed with repeated showers in warmer water (15 degrees C). It is concluded that when the body surface area cooled is the same, the rate of change of skin temperature is an important factor in determining the degree of habituation produced. PMID- 15778889 TI - Functional magnetic resonance imaging studies of opioid receptor-mediated modulation of noxious-evoked BOLD contrast in rats. AB - RATIONALE: Functional magnetic resonance imaging (fMRI) in rats can non invasively identify brain regions activated by physiological stimuli and the effects of pharmacological intervention on these responses. OBJECTIVES: This study was conducted to investigate the effects of systemic administration of the mu-opioid receptor agonist morphine on whole brain functional signal intensity in anaesthetised rats; to investigate whether pre-treatment with the opioid receptor antagonist naloxone blocks the effects of morphine; to determine whether pre treatment with morphine attenuates noxious-evoked changes in whole brain functional signal intensity. METHODS: Continuous whole brain fMRI scanning was used to study brain signal intensity prior to, and following, systemic administration of morphine (5 mg/kg, n=7), systemic administration of naloxone (1 mg/kg) and morphine (n=8). Effects of pre-treatment with saline (n=5) or morphine (5 mg/kg, n=5) on formalin (5%, intraplantar)-evoked changes in signal intensity were determined. Data were processed using SMP99 with fixed-effects analysis (p<0.05). RESULTS: Morphine produced significant positive bilateral increases in signal intensity in the cingulate cortex, amygdala, thalamus, hypothalamus and PAG (p<0.05), and these effects were blocked by naloxone. Intraplantar injection of formalin produced a significant positive increase in signal intensity in the cingulate cortex, somatosensory cortex, amygdala, thalamus, hypothalamus and PAG (p<0.05). Morphine attenuated formalin-evoked increases in signal intensity in the PAG, amygdala, hypothalamus and cingulate cortex. CONCLUSION: Our data demonstrate that morphine modulates noxious-evoked changes in signal intensity in discrete brain regions. fMRI studies in rats are able to identify specific brain regions involved in the pharmacological modification of physiologically evoked changes in regional brain activation. PMID- 15778891 TI - Motivational aspects of maternal anxiolysis in lactating rats. AB - RATIONALE AND OBJECTIVE: This study examines the role of maternal motivation on the reduced anxiety-like responses displayed by lactating rats in the plus maze test. RESULTS: Maternal animals, both lactating and sensitized (ovariectomized females behaving maternal after a continuous exposure to pups), displayed anxiolytic-like responses in the plus maze test in contrast to ovariectomized non maternal rats. However, the levels of experimental anxiety were lower in lactating than in sensitized females. Pups placed in the open arms of the maze further reduced the low levels of anxiety-like behavior of both sensitized and lactating rats. Low doses of haloperidol (0.05 and 0.1 mg/kg), a dopamine antagonist, which interfere with maternal motivation but has neither anxiolytic nor anxiogenic effect in the plus maze test, significantly increased the anxiety like responses of lactating rats. The presence of the pups in the open arms of the maze overrode the behavioral effect of haloperidol on lactating dams' anxiety related behavior. CONCLUSIONS: These experiments show that maternity induces changes in the way the animals react to the environment, rendering them less anxious to aversive stimuli. The degree of experimental anxiolysis displayed by maternal animals varies according to their maternal motivation, which is modulated by the female's endocrine state, the pups and/or the dopaminergic system. PMID- 15778893 TI - Sustained hyperglycaemia increases muscle blood flow but does not affect sympathetic activity in resting humans. AB - An increase in capillary blood flow and pressure in response to diabetes mellitus may lead to microangiopathy. We hypothesize that these haemodynamic changes are caused by a decreased activity of the sympathetic nervous system due to episodes of sustained hyperglycaemia. Twelve healthy volunteers consecutively underwent a hyperglycaemic experiment (HYPER), with the plasma glucose level maintained at 20 mmol.l(-1) for 6 h by combined infusion of somatostatin, insulin and glucose; and a normoglycaemic experiment (NORMO), with similar infusions but with the plasma glucose maintained at fasting level. During both experiments, sympathetic nervous system (SNS) activity was measured by assessing the plasma catecholamine levels, microneurography, power spectral analysis and forearm blood flow (FBF). In an age and weight matched group, fasting and 6-h sympathetic activity was measured without infusion of somatostatin and insulin (CONTROL). During HYPER, forearm blood flow increased from 2.45 (0.21) to 3.10 (0.48) ml.dl(-1).min(-1) ( P <0.05), but did not change in NORMO or CONTROL. The HYPER conditions did not change the plasma noradrenaline levels or the muscle sympathetic nerve activity [42 (4), 50 (10) and 45 (5) bursts/100 beats, HYPER, NORMO and CONTROL respectively]. Also, the power spectral analysis was similar under all experimental conditions. All results are expressed as the mean (SEM). In conclusion, sustained hyperglycaemia in normal subjects induces moderate vasodilation in skeletal muscle, but this increased blood flow can not be attributed to a decreased sympathetic tone. PMID- 15778890 TI - Methodological considerations in rat brain BOLD contrast pharmacological MRI. AB - RATIONALE AND OBJECTIVES: Blood oxygen level dependent (BOLD) contrast pharmacological magnetic resonance imaging (phMRI) is an increasingly popular technique that allows the non-invasive investigation of spatial and temporal changes in rat brain function in response to pharmacological stimulation in vivo. Rat brain BOLD contrast phMRI is, at present, established in few neuropharmacological laboratories, and various issues associated with the technique require attention. The present review is primarily aimed at psychopharmacologists with no previous experience of phMRI, who are interested in the practical aspects that phMRI studies entail. RESULTS AND DISCUSSION: Experimental and analytical considerations, including anaesthesia, physiological monitoring, drug dose and delivery, scanning protocols, statistical approaches and the interpretation of phMRI data, are discussed. PMID- 15778894 TI - Effects of voluntary wheel running and amino acid supplementation on skeletal muscle of mice. AB - The aims of the present study were as follows: (1) to examine the adaptational changes to chronic endurance voluntary exercise and (2) to investigate the effects of amino acid supplementation on the adaptational changes induced by endurance training in hindlimb (gastrocnemius, tibialis, soleus) and respiratory (diaphragm) muscles of mice. Male C57Bl6 mice were divided in four groups: control sedentary, sedentary supplemented with amino acid mixture (BigOne, 1.5 mg g day(-1) in drinking water for 8 weeks), running (free access to running wheels for 8 weeks), and running supplemented with amino acid mixture. Myosin heavy chain (MHC) isoform distribution was determined in all muscles considered. Fiber cross-sectional area (CSA) was measured in the soleus muscle. In all muscles except the tibialis, endurance training was associated with an overall shift towards the expression of slower MHC isoforms. Amino acid supplementation produced a shift towards the expression of faster MHC isoforms in the soleus and diaphragm muscles, and partially antagonized the effects of training. Immunohistochemical analysis of CSA of individual muscle fibers from the soleus muscle suggests that voluntary running produced a decrease in the size of type 1 fibers, and amino acid supplementation during training resulted in an increase in size in both type 1 and type 2A fibers. Collectively, these results suggest that the endurance adaptations induced by voluntary running depend on the muscle type, and that amino acid supplementation is able to modulate both fiber size and MHC isoform composition of skeletal muscles in sedentary and exercised mice. PMID- 15778895 TI - Salivary IgA response to prolonged exercise in a hot environment in trained cyclists. AB - The aim of this study was to determine the effects of prolonged exercise in hot conditions on saliva IgA (s-IgA) responses in trained cyclists. On two occasions, in random order and separated by 1 week, 12 male cyclists cycled for 2 h on a stationary ergometer at 62 (3)% V(.)O(2 max) [194 (4) W; mean (SEM)], on one occasion (HOT: 30.3 degrees C, 76% RH) and on another occasion ( CONTROL: 20.4 degrees C, 60% RH). Water was available ad-libitum. Venous blood samples and 2 min whole unstimulated saliva samples were collected at pre, post and 2 h post exercise. The s-IgA concentration was determined using a sandwich-type ELISA. Exercising heart rate, rating of perceived exertion, rectal temperature, corrected body mass loss (P<0.01) and plasma cortisol (P<0.05) were greater during HOT. The decrease in plasma volume post-exercise was similar on both trials [HOT: -6.7 (1.1) and CONTROL: -6.6 (1.3)%; P<0.01]. Saliva flow rate decreased post-exercise by 43% returning to pre-exercise levels by 2 h post exercise (P<0.05) with no difference between trials. Saliva IgA concentration increased post-exercise (P<0.05) with no difference between trials. Saliva IgA secretion rate decreased post-exercise by 34% returning to pre-exercise levels by 2 h post-exercise (P<0.05) with no difference between trials. These data show that a prolonged bout of exercise results in a reduction in s-IgA secretion rate. Additionally, these data demonstrate that performing prolonged exercise in the heat, with ad libitum water intake, does not influence s-IgA responses to prolonged exercise. PMID- 15778896 TI - Carbohydrate supplementation and alterations in neutrophils, and plasma cortisol and myoglobin concentration after intense exercise. AB - The present study examined the effect of carbohydrate supplementation on changes in neutrophil counts, and the plasma concentrations of cortisol and myoglobin after intense exercise. Eight well-trained male runners ran on a treadmill for 1 h at 85% maximal oxygen uptake on two separate occasions. In a double-blind cross over design, subjects consumed either 750 ml of a 10% carbohydrate (CHO) drink or a placebo drink on each occasion. The order of the trials was counter-balanced. Blood was drawn immediately before and after exercise, and 1 h after exercise. Immediately after exercise, neutrophil counts (CHO, 49%; placebo, 65%; P<0.05), plasma concentrations of glucose (CHO, 43%; P<0.05), lactate (CHO, 130%; placebo, 130%; P<0.01), cortisol (CHO, 100%; placebo, 161%; P<0.01), myoglobin (CHO, 194%; placebo, 342%; P<0.01) all increased significantly. One hour post-exercise, plasma myoglobin concentration (CHO, 331%; placebo, 482%; P<0.01) and neutrophil count (CHO, 151%; placebo, 230% P<0.01) both increased further above baseline. CHO significantly attenuated plasma myoglobin concentration and the neutrophil count after exercise (P<0.01), but did not affect plasma cortisol concentration. The effects of CHO on plasma myoglobin concentration may be due to alterations in cytokine synthesis, insulin responses or myoglobin clearance rates from the bloodstream during exercise. Plasma cortisol responses to CHO during exercise may depend on the intensity of exercise, or the amount of CHO consumed. Lastly, cortisol appears to play a minor role in the mobilisation of neutrophils after intense exercise. PMID- 15778898 TI - Peak oxygen uptake during running and arm cranking normalized to total and regional skeletal muscle mass measured by magnetic resonance imaging. AB - The purposes of our study were to determine the peak oxygen uptake (V(.) >O(2peak)) per total or regional skeletal muscle (SM) mass using magnetic resonance imaging (MRI) and to investigate the relationships between SM mass and V(.) >O(2peak) during running and arm cranking. Eight male college swimmers aged 18-22 years [mean (SD) age 20.0 (1.3) years] were recruited to participate in this study. V(.) >O(2) during running and arm cranking were measured using an automated breath-by-breath mass spectrometry system. Contiguous MRI slices were obtained from the first vertebra cervicale to the malleolus lateralis (1.0-cm slice thickness, 0-cm inter-slice gap), resulting in a total of approximately 156 images for each subject. The absolute V(.) O(2peak) and the V(.) O(2peak) per body mass during running and arm cranking were 3.6 (0.6) l.min(-1), 54.4 (5.9) ml.min(-1).kg(-1) and 2.5 (0.5) l.min(-1), 36.9 (5.3) ml.min(-1).kg(-1), respectively. The absolute V(.) O(2peak) was higher ( P <0.05) during running than during arm cranking, but not the V(.) O(2peak) per regional area SM mass. The lower body SM mass was correlated to the V(.) O(2peak) during running ( r =0.95, P <0.001). All measurements and calculated values were expressed as the mean (SD) for the eight subjects. To eliminate the influence of body mass and fat free mass (FFM), a regression analysis was performed on the mass-residuals of the V(.) O(2peak) during running and the lower body SM mass. The residuals of lower body SM mass were correlated to the residuals of V(.) O(2peak) during running, with respect to body mass ( r =0.90, P <0.001) and FFM ( r =0.82, P <0.05). These results suggest that the MRI-measured lower body SM mass was closely associated to the absolute V(.) O(2peak) during running, independently of body mass or FFM, and that the V(.) O(2peak) per regional SM mass corresponded, regardless of the type of exercise (upper or lower body). PMID- 15778897 TI - Differential responses of serum and salivary interleukin-6 to acute strenuous exercise. AB - Physical exercise is associated with elevation of serum levels of interleukin-6 (IL-6) because of its production in the muscles. The use of IL-6 measurements in saliva has been proposed in the field of immunopathology, mainly involving salivary gland disease. We evaluated the responses of serum and salivary IL-6 in two different groups of athletes submitted to different types of controlled strenuous exercise (spinning activity and maximal isokinetic test). Serum and salivary samples for IL-6 measurements, and serum samples for lactate and myoglobin determination before and after exercise, were obtained. Salivary IL-6 was measured by ELISA after dilution experiments and compared with results obtained by immunoradiometric assay. Spinning activity elicited significant increases in all the variables, and no correlation was found among the respective variations. A significant response to the isokinetic exercise was observed for serum IL-6, lactate and myoglobin only; no correlation was found between serum and salivary IL-6. Our study demonstrated that serum and salivary IL-6 responses to exercise are dissociated, possibly in relation to the lack of relationships between the systemic/muscular and the salivary routes of IL-6 production. Analytical issues that concern IL-6 measurement in saliva deserve attention, notably regarding the collection method used to absorb saliva. Concomitant monitoring of serum markers of inflammation, muscle metabolism and damage can provide information about muscle function properties and adaptations to physical effort in different types of athletes. PMID- 15778899 TI - Influence of different rest intervals during active or passive recovery on repeated sprint swimming performance. AB - The purpose of this study was to investigate the effect of active or passive recovery after two different rest intervals on performance during repeated bouts of maximal swimming exercise. Sixteen swimmers (eight males and eight females) performed four trials in a counterbalanced order. Eight repetitions of 25-m sprints (8 x 25 m), with a rest interval of 45 or 120 s, followed by a 50-m sprint test 6 min later, were performed in each trial. The 45 or 120-s interval was either active (A45 and A120) or passive (P45 and P120). The intensity of the active recovery corresponded to 60% of the individual best 100-m velocity. Performance time was recorded using an official competition timing system. The first 25-m sprint was comparable across trials (P>0.05), but performance was decreased after the second sprint during active compared to passive recovery, irrespective of the interval duration (P<0.05). The 50-m sprint time was 2.4% better in the P120 and A120 compared to the A45 and P45 trials (P<0.05). After completing the 8x25 m, blood lactate was decreased with active recovery when the interval period was 120 s (P120 vs A120, P<0.05). Blood lactate concentration at the start as well as 5 min after the 50-m sprint was lower in the A120 and A45 compared to the P120 and P45 trials respectively (P<0.05). Plasma glycerol was not different between trials (P>0.05), whereas plasma ammonia was higher in the A45 compared to the P120 trial (P<0.05). The interval period separating short duration sprints may therefore alter performance when subsequent maximum exertion is applied. For sustained sprinting ability, passive recovery is advised during repeated swimming sprints of short duration. PMID- 15778900 TI - Aggravated hypoxia during breath-holds after prolonged exercise. AB - Hyperventilation prior to breath-hold diving increases the risk of syncope as a result of hypoxia. Recently, a number of cases of near-drownings in which the swimmers did not hyperventilate before breath-hold diving have come to our attention. These individuals had engaged in prolonged exercise prior to breath hold diving and it is known that such exercise enhances lipid metabolism relative to carbohydrate metabolism, resulting in a lower production of CO(2) per amount of O(2 )consumed. Therefore, our hypothesis was that an exercise-induced increase in lipid metabolism and the associated reduction in the amount of CO(2) produced would cause the urge to breathe to develop at a lower P O(2), thereby increasing the risk of syncope due to hypoxia. Eight experienced breath-hold divers performed 5 or 6 breath-holds at rest in the supine position and then 5 or 6 additional breath-holds during intermittent light ergometer exercise with simultaneous apnoea (dynamic apnoea, DA) on two different days: control (C) and post prolonged sub-maximal exercise (PPE), when the breath-holds were performed 30 min after 2 h of sub-maximal exercise. After C and before the prolonged submaximal exercise subjects were put on a carbohydrate-free diet for 18 h to start the depletion of glycogen. The respiratory exchange ratio ( RER) and end tidal P CO(2), P O(2), and SaO(2) values were determined and the data were presented as means (SD). The RER prior to breath-holding under control conditions was 0.83 (0.09), whereas the corresponding value after exercise was 0.70 (0.05) ( P <0.01). When the three apnoeas of the longest duration for each subject were analysed, the average duration of the dynamic apnoeas was 96 (14) s under control conditions and 96 (17) s following exercise. Both P O(2) and P CO(2) were higher during the control dynamic apnoeas than after PPE [PO(2) 6.9 (1.0) kPa vs 6.2 (1.2) kPa, P <0.01; P CO(2) 7.8 (0.5) kPa vs 6.7 (0.4) kPa, P <0.001; ANOVA testing]. A similar pattern was observed after breath-holding under resting conditions, i.e., a lower end-tidal P O(2) and P CO(2) after exercise (PPE) compared to control conditions. Our findings demonstrate that under the conditions of a relatively low RER following prolonged exercise, breath-holding is terminated at a lower P O(2) and a lower P CO(2) than under normal conditions. This suggests that elevated lipid metabolism may constitute a risk factor in connection with breath-holding during swimming and diving. PMID- 15778901 TI - Evaluation of microsatellite markers in association studies: a search for an immune-related susceptibility gene in sarcoidosis. AB - Association studies using linkage disequilibrium (LD) between candidate loci and nearby markers have been proposed to identify susceptibility genes for complex diseases. We analyzed polymorphisms of microsatellites (MSs) and LD patterns of the regions in which candidate genes related to the Th1 immune response have been annotated and attempted to identify a susceptibility gene for sarcoidosis in a marker-based association study. Nineteen MSs were identified in six Th1-related genes (IFNGR1, IFNGR2, IL12RB1, IL12RB2, STAT1 and STAT4) and then eight were further characterized as useful polymorphic markers. Most of these MSs showed LD with single nucleotide polymorphisms (SNPs) on both 5' and 3' ends of these candidate genes, in which r(2) values between at least one of the MS marker alleles and the SNPs were higher than 0.1. A significant association with one MS allele near STAT4 was shown and a cluster of SNPs in LD with the MS marker was associated with sarcoidosis. These results suggest that association studies using not only SNPs but also multi-allelic MS within or near candidate loci would be useful markers to search for a disease susceptibility gene, especially in populations with unknown LD structure. PMID- 15778902 TI - Structure and the evolutionary implication of the triplicated complement factor B genes of a urochordate ascidian, Ciona intestinalis. AB - To elucidate the evolution of the complement system and MHC class III region, we analyzed the complement factor B (Bf) genes of a urochordate ascidian, Ciona intestinalis. Three different cDNA species, termed CiBf-1, CiBf-2 and CiBf-3, were identified. The deduced amino-acid sequences all contained the usual domains of vertebrate Bf and, in addition, three extra domains at the N-terminus. Furthermore, the serine protease domain of these CiBfs shared unique features with vertebrate complement components C1r/s and mannose-binding lectin-associated serine protease (MASP)-2/3, the absence of the disulfide bond designated histidine loop, and the usage of the AGY codon for the catalytic serine residue. These results indicate that complement genes have evolved through extensive exon shuffling events in the early stage of chordate evolution. Overall deduced amino acid identity between CiBf-1 and -2 was 88%, whereas CiBf-3 showed 49% identity to both CiBf-1 and CiBf-2. These three CiBf genes were located within an approximately 50-kb genomic region, and exons 3 and 5 of all the three Bf genes showed an extremely high degree of nucleotide identity, indicating that the CiBf genes experienced extensive reorganization, such as duplication and gene conversion, since its divergence from the vertebrate Bf/C2 gene. Fluorescent in situ hybridization (FISH) to the chromosomes showed that genetic loci for the CiBfs, CiC3-1 and CiC3-2 genes are present on three different chromosomes, suggesting the possibility that the linkage among the MHC class III complement genes was established in the vertebrate lineage after its divergence from urochordates. PMID- 15778905 TI - Interobserver reliability of ICSD-R minimal diagnostic criteria for the parasomnias. AB - OBJECTIVE: To estimate the interobserver reliability (IR) of the minimal diagnostic criteria for the parasomnias provided in the International Classification of Sleep Disorders Revised (ICSD-R). METHODS: Fifty consecutive subjects underwent a structured interview by three doctors based on the ICSD-R minimal criteria for the diagnosis of 13 parasomnias at any time in life. IR was calculated by Kappa statistics and interpreted according to conventional categories. RESULTS: In the group of Arousal Disorders, IR was "substantial" (kappa 0.74) for confusional arousals, "slight" (kappa 0.36) for sleepwalking, "fair" (kappa -0.02) for sleep terrors. In the group of Wake-Sleep Transition Disorders, IR was "substantial" to "almost perfect", but "moderate" for sleep starts (kappa 0.41). In the group of Parasomnias usually associated with REM Sleep, IR was "substantial" (kappa 0.69) for sleep paralysis, "moderate" (kappa 0.46) for RBD, "fair" (kappa 0.25) for nightmares. In the group of Other Parasomnias, IR was "substantial" to "almost perfect" (kappa between 0.73 and 0.93). CONCLUSIONS: When the clinical diagnosis of parasomnias is based on the ICSD-R: 1) the majority of Arousal Disorders and REM sleep parasomnias showed only a "fair" to "moderate" IR; 2) all of the other parasomnias showed a "substantial" to "almost perfect" IR. Nosological entities with unsatisfactory IR share complex motor phenomenology associated with a mental state difficult for the patient to define. The source of disagreement probably lies in the difficulty in interpreting patients' reports. For these parasomnias IR must be verified and possibly improved with training. PMID- 15778904 TI - The receptor mechanism mediating the contractile response to adenosine on lung parenchymal strips from actively sensitised, allergen-challenged Brown Norway rats. AB - Parenchymal strips prepared from lungs removed from actively sensitised Brown Norway rats challenged with allergen show hyperresponsiveness to adenosine. The response is mast cell mediated and a preliminary pharmacological analysis suggested the involvement of a receptor (or receptors) that could not be classified as any of the known adenosine receptor subtypes. We present a further analysis of the response. Male Brown Norway (BN) rats, actively sensitised to ovalbumin (OA), were challenged intratracheally with OA and killed 3 h later to provide parenchymal strip preparations. The augmented contractile responses to adenosine were partially blocked by the 5-HT receptor antagonist, methysergide, or the A(1) receptor antagonist, DPCPX, and abolished in the presence of both antagonists. Responses to high concentrations of the A(1) receptor agonist, CPA were, like those to adenosine, augmented on tissues from allergen-challenged animals and blocked by a combination of methysergide and DPCPX. The A(3) receptor agonist, Cl-IB-MECA, did not contract the tissue, but partially blocked the response to adenosine. A combination of Cl-IB-MECA and methysergide induced a similar degree of blockade to that seen with either drug given alone. Combination of Cl-IB-MECA and/or methysergide with DPCPX abolished the response to adenosine. The effects of the A(3) receptor agonist, inosine, were augmented on tissues from allergen-challenged animals and markedly inhibited by disodium cromoglycate, methysergide or Cl-IB-MECA. Responses to adenosine were abolished when parenchymal strips were taken from rats pretreated 48 h previously with pertussis toxin. 8-SPT, CGS 15943, XAC, MRS 1754, DPCPX and theophylline, at concentrations which inhibit the A(1) A(2A) and/or A(2B) receptors but have negligible affinity for the rat A(3) receptor, inhibited responses to adenosine, but high concentrations were required and blockade was incomplete. MRS 1523 and MRS 1191, which are antagonists at the rat A(3) receptor, had no effect on the response to adenosine. The present results support and clarify our earlier conclusion that an atypical receptor mechanism mediates contraction of the parenchymal strip prepared from the lungs of actively sensitised BN rats challenged with allergen to adenosine. The response arises from a combined effect of adenosine on the A(1) receptor and a receptor with similarities to the A(3) receptor, but where Cl-IB MECA behaves as an antagonist and MRS 1523 and MRS 1191 are inactive at concentrations that substantially exceed their affinities for the rat A(3) receptor. PMID- 15778906 TI - Vermal atrophy of alcoholics correlate with serum thiamine levels but not with dentate iron concentrations as estimated by MRI. AB - Chronic alcohol consumption is frequently accompanied by cerebellar degeneration. The exact aetiology of alcoholic cerebellar degeneration is still a matter of debate. The aim of the present study was to investigate whether patients with chronic alcohol consumption exhibit a decrease in dentate nuclei intensity as measured by MRI, and if so, whether this decrease correlates with cerebellar atrophy as revealed by MR imaging or with clinical signs of cerebellar ataxia. A decrease in dentate nuclei intensity would indirectly indicate that iron accumulation, and therefore, oxidative stress may play a role in alcoholic cerebellar degeneration. MRI of 45 alcoholics and 44 age and sex-matched healthy control subjects was performed using a 3D-T1-weighted fast low angle shot (FLASH) echo sequence. Signal intensities of the dentate nuclei and cerebellar white matter were bilaterally measured. Planimetric measurements of cerebellar size were performed using a 3D-T1-weighted magnetization prepared rapid acquisition gradient echo (MPRAGE) sequence. Results demonstrated that dentate nuclei intensity was not significantly decreased in patients with chronic alcohol consumption (mean +/- SD signal intensity 65.36 +/- 13.0) if compared with control subjects (mean +/- SD signal intensity 68.95 +/- 9.4) (p = 0.15). Dentate nuclei intensity did not correlate with cerebellar size neither in control subjects nor in alcoholics. In contrast, vitamin B1 level correlated with cerebellar size in alcoholics even if the vitamin B1 concentration was within normal values (r = 0.344, p = 0.028). These results support the view that thiamine deficiency rather than direct neurotoxic effects of alcohol is the main causative factor for the development of alcoholic cerebellar degeneration. PMID- 15778908 TI - Acute peripheral facial palsy in adults. AB - OBJECTIVE: To collect epidemiological data of peripheral facial palsy, and especially to chart the incidence and clinical characteristics of Lyme associated facial palsy. MATERIAL AND METHODS: We included consecutive adult patients presenting with peripheral facial palsy in Vest-Agder County from January 1997 to December 1998. The facial palsy was graded according to the House and Brackman facial function scoring system,and cerebrospinal fluid and serum samples were examined for Borrelia burgdorferi antibodies and virus antibodies. Final outcome was evaluated by follow up visits or telephone interview. RESULTS: Sixty nine patients were included and followed until complete recovery, or for 5 years. Ten per cent were caused by Lyme disease, 17% by virus infection, 4% by other causes and 68% were classified as Bell's palsy. All patients with Lyme facial palsy had additional neurological symptoms, and 87% reported constitutional complaints. The overall final outcome was good with complete recovery in 77%, slight sequelae in 20% and moderate sequelae in 3%. No patients experienced severe sequelae. Two of 28 patients examined with neurography had absent compound muscle action potentials in orbicularis oculi. Both made good recovery with only slight sequelae. CONCLUSIONS: Peripheral facial palsy is a common disorder with a favourable prognosis. Lyme disease seems to be an infrequent cause of facial palsy in patients without constitutional symptoms or additional neurological findings. PMID- 15778907 TI - Postherpetic neuralgia: topical lidocaine is effective in nociceptor-deprived skin. AB - OBJECTIVES: Topical lidocaine is effective in postherpetic neuralgia (PHN). The aim of the present investigation was to classify patients according to their predominant peripheral nociceptor function and to compare these data with the results of a controlled study using dermal lidocaine patch. METHODS: Within the skin area of maximal pain QST (thermotest) and QCART (histamine iontophoresis and laser Doppler flowmetry) were performed prospectively in 18 PHN patients. A controlled study using cutaneous lidocaine (lidocaine 5% patch, IBSA) followed. RESULTS: Six patients (group I, sensitised nociceptors) had no sensory loss. Heat pain thresholds were equal or lower than on the contralateral side. Histamine induced flare and axon reflex vasodilatation were not different on both sides. Histamine evoked pain increased. In 12 patients (group II, nociceptor impairment) heat pain thresholds were higher than contralateral. Histamine-induced flare was impaired or abolished. Histamine did not induce any sensation. Lidocaine was efficacious in the entire group of patients. Subgroup analysis revealed that patients with impairment of nociceptor function had significantly greater pain reduction under lidocaine vs placebo. Patients with preserved and sensitised nociceptors demonstrated no significant pain relief. CONCLUSIONS: PHN patients differ concerning their cutaneous nociceptor function: In the group I pain is caused by pathologically sensitised nociceptors. In subset II there is a loss of function of cutaneous C-nociceptors within the allodynic skin. Patients responded well to topical lidocaine even if the skin was completely deprived of nociceptors. Different underlying mechanisms of lidocaine action in nociceptor deprived skin are discussed. PMID- 15778909 TI - Only physical aspects of quality of life are significantly improved by bilateral subthalamic stimulation in Parkinson's disease. AB - BACKGROUND: The well known global improvement of quality of life (QoL) after bilateral high frequency chronic deep brain stimulation of the subthalamic nucleus (STN DBS) in Parkinson's disease (PD) is in contrast to behavioral disturbances as observed after surgery. Indeed the impact of DBS on physical versus mental aspects of QoL in PD remains unknown. OBJECTIVE: To assess the influence of bilateral STN DBS on physical versus mental aspects of QoL in Parkinson's disease. METHODS: The results of 27 patients for the Unified Parkinson's disease Rating Scale (UPDRS), Parkinson's Disease Questionnaire 39 (PDQ39) and Short Form 36 health survey questionnaire (SF36) were compared before surgery and after 12 months of bilateral STN DBS. RESULTS: Comparing off-dopa conditions before versus 12 months after surgery, both UPDRS part II and part III significantly improved: 32.6% and 52%, respectively. UPDRS part I scores did not change significantly at 12 months. As for PDQ39, the global score significantly improved after surgery (21.1 %) as did four subscores: mobility (25.6 %), activity of daily living (34.5 %), stigma (40.1 %) and bodily discomfort (30 %). Three PDQ39 subscores, however, showed no significant changes: emotional well being (10.7 %), social support (3.2%) and cognition (8.5 %) and one item even worsened: communication (-7.7 %). In SF36, only physical items significantly improved. CONCLUSION: Using clinician's based rating scale, bilateral STN DBS showed significant improvement in PD patients at 12 month follow up. However, using patient's self-assessment scales, the clinical benefit of STN DBS was more subtle: physical items of QoL significantly improved, whereas mental items such as emotional well-being, social support, cognition and communication showed no improvement. Our results are suggestive of a dissociation of motor and non-motor symptoms control after bilateral STN DBS in PD patients. PMID- 15778910 TI - Measuring carotid artery stenosis--comparison of postmortem arteriograms with the planimetric gold standard. AB - OBJECTIVE: Five different calliper methods for assessing the degree of carotid artery stenosis and visual estimation ("eyeballing") of postmortem carotid arteriograms were compared with the planimetric gold standard of the area reduction at the site of the stenosis. METHODS: During autopsy 53 carotid specimens were removed in toto from 31 neurological patients. Carotid arteries were ligated and redistended to a physiological degree for standardised three plane arteriography. Afterwards, the entire specimen was filled with an embedding medium under the same conditions and sectioned. Slices at the site of stenoses were histologically processed. Computerised planimetric analysis of the lumen area reduction was performed and compared with the arteriographic findings. Arteriograms were evaluated by two independent observers by means of linear Common Carotid Artery (CC), the European Carotid Surgery Trial (ECST) and the North American Symptomatic Carotid Endarterectomy Trial (NASCET), and squared measurements (NASCET2, ECST2) after applying the pi r2 function. Further, three independent observers performed eyeballing of the degree of stenosis from the postmortem arteriographies. RESULTS: Planimetry was carried out in 29 internal carotid artery (ICA) and 17 common carotid artery (CCA) stenoses ranging from 8.5 to 100%. The smallest mean differences of the degree of stenosis in % between planimetry and arteriography were -0.5 and 0.6%. The narrowest 95 %-limits of agreement covered a range of +/-24.1 and 26.3% of stenoses, and the highest correlation coefficients were both 0.9 for the CC and ECST2 techniques, respectively. By eyeballing, the degree of stenosis was underestimated by 13.5 to 15.8% on average. The narrowest limits of agreement between two observers for eyeballing covered a range of 35%. CONCLUSION: Three-plane arteriography has only a moderate accuracy and reproducibility in detecting and measuring carotid artery stenosis independent of the technique of measurement used. PMID- 15778912 TI - The auricle in the visual arts. PMID- 15778913 TI - Incisionless otoplasty. AB - Otoplasty surgery for correction of the "lop" protuberant ear deformity continues to evolve. A noninvasive method for achieving normal appearance and physiology for the protuberant ear pinna is presented. Incisionless Otoplasty surgery, involving a combined cartilage scoring procedure and a percutaneously placed retention suture procedure, creates a predictable and permanent correction of the missing antihelical fold. In addition, treatment of the protruding conchal bowl by endoscopic techniques further enhances the cosmetic appearance. A long-term follow-up period of over 10 years gives perspective on the efficacy of the operation. PMID- 15778914 TI - The modified converse technique in combination with cavum rotation. AB - We report our experiences in 210 patients with protruding ears who underwent a surgical correction of their deformity using a modified "Converse technique." The technique combines a posterior incomplete cartilage incision with permanent cartilage sutures and a cavum rotation if necessary. The described technique provides a good anterior width and normal relief, with shaped anthelix and crus superior, can be achieved. PMID- 15778915 TI - Otoplasty: a modified anterior scoring technique. AB - In the long history of otoplasty, the basic techniques used to correct prominent ears with a missing antihelix are based on incisions, scoring, and suturing of the cartilaginous framework of the auricle. In this article, modifications (subperichondrial dissection and adjusting sutures) of the anterior scoring technique of Chongchet are described. PMID- 15778916 TI - The incision-excision technique in minor auricular deformities. AB - In 1972, in the search for a method to cope with all deformities in protruding ears and other common minor auricular deformities, Claus Walter published a surgical technique based on various incisions and excisions of the cartilage. The procedure consists of a complete separation of the auricle into two parts, combined with remodeling of antihelical, helical, and lobular structures without use of adjusting sutures. Thus, even in patients with stiff and unyielding cartilage and in revisional surgery, pleasing aesthetic results may be achieved. The recurrence rate has found to be reduced markedly. Considering all pros and cons and optional surgical modifications, the incision-excision technique can be adopted for every protruding ear as well as for lop ears, moderate cup ear deformities, and secondary revisions. Because of the extent of cartilage dissection, this technique is not recommended for beginners but should be restricted to experienced surgeons with profound knowledge of basic remodeling procedures. PMID- 15778917 TI - Correction of the lobule. AB - Many techniques have been described for the correction of protruding ears. Most of them concentrate on correcting the form and position of auricular cartilage. The lobule is a soft tissue structure. Skin resections of its posterior surface have been propagated for the correction of its position; however, these cause tension on the wound and might increase the already relatively high risk for the development of keloids. We have modified the technique for correcting the protruding lobule for its exact positioning and minimizing the risk for relapse and keloids. Starting from the incision performed for the anthelix plasty, a subcutaneous pocket is prepared between the anterior and posterior sides of the lobule. Afterwards, the subcutaneous layer of the postlobular skin is adjusted and fixed to the cartilage of the conchal cavum with a special mattress suture. This technique is a refinement of otoplasty for bat ears. It is indicated for precise modification of form and position of protruding lobules. PMID- 15778918 TI - Synopsis of otoplasty. PMID- 15778919 TI - RNA interference-based strategies for metabolic syndrome treatment. AB - RNA interference is a naturally occurring cellular mechanism to inhibit the expression of specific gene products. The technical application of RNA interference offers great potential for the specific treatment of a huge variety of diseases including the metabolic syndrome, one of the most challenging threats to human health associated with our civilization. In order to develop novel and powerful strategies for the treatment of the metabolic syndrome, it is essential to define a set of specific gene products that may be targeted by RNA interference. Based on currently available in vitro and in vivo data, we discuss the feasibility of candidate genes involved in the pathophysiology of the metabolic syndrome as potential targets for a rational RNA interference based therapy in this review. PMID- 15778920 TI - Transfer of diabetes from prediabetic NOD mice to NOD-SCID/SCID mice: association with pancreatic insulin content. AB - Splenocytes from prediabetic female NOD mice can transfer diabetes to NOD-SCID mice. Whereas the kinetics of disease transfer was shown to be a function of the age of donor splenocytes, information is scarce as to how the stage of autoimmune disease, as evaluated by pancreatic insulin content, is related to the diabetogenic potency of splenic T-cells. We therefore determined individual diabetes transfer times after an i. v. injection of splenocytes from prediabetic NOD mice of different ages into female NOD-SCID mice in relation to the diabetes incidence in NOD donor mice and their pancreatic insulin contents. Three groups (n = 8) of NOD mice aged 5, 11, and 17 weeks (wk) underwent splenectomy and hemipancreatectomy. After that, 10x10 (6) splenocytes either pooled from all donor NOD mice of the different age groups or individually from single donor mice were transferred to groups of four 6-week-old NOD-SCID mice, respectively, in two sets of experiments. Insulin was extracted from the resected hemipancreas, and the insulin content was determined by a RIA. Diabetes in the NOD-SCID cohort occurred after a mean time of 126 days after transfer of pooled splenocytes from 5-week-old NODs, after 68 days (transfer from 11-week-old NODs), and after a mean time of 43 days (transfer from 17-week-old NODs, 5 vs. 11 wk: p < 0.02, 11 vs. 17 wk: p < 0.001). Individual time to diabetes positively correlated with diabetes transfer times in NOD-SCID recipients (p < 0.0001) in the 17-week-old NOD mice, confirming previous diabetes transfer studies in hemi-pancreatectomized NOD mice. Furthermore, individual insulin concentrations in 17-week-old NOD mice also positively correlated to diabetes transfer times in recipient mice (p < 0.0001). No such correlations for these parameters were seen for the 5 and 11-week-old NOD mice (time to diabetes: 11 wk, p = 0.14, 5 wk, p = 0.75; insulin content: 11 wk, p = 0.81, 5 wk, p = 0.14). These data suggest that destructive T-cell activity increases during the course of islet autoimmunity. The immune response seems to be programmed for beta-cell destruction just before diabetes onset. This is the only time that pancreatic insulin content predicts the impending onset of diabetes. PMID- 15778921 TI - Blood flow in subcutaneous adipose tissue depends on skin-fold thickness. AB - Blood flow in subcutaneous adipose tissue is reduced in obese compared to lean subjects. Limitations in vascular supply might interfere with adipose tissue function as a metabolic and endocrine organ. We tested the hypothesis that nutritive blood flow and tissue metabolism depends on subcutaneous adipose tissue thickness even in normal-weight subjects. Sixteen young, healthy, normal-weight subjects (8 men, 8 women) were included in the study. Abdominal subcutaneous adipose thickness was assessed by skin-fold measurements. The microdialysis technique was applied for monitoring basal adipose tissue blood flow (ethanol dilution technique) and metabolism. An increase in skin-fold thickness from 15 to 45 mm and from 8 to 37 mm was associated with a linear increase in basal ethanol ratio from 0.19 to 0.63 and 0.25 to 0.75 and linear decreases in dialysate glucose concentrations from 1.95 to 0.24 mM and 1.68 to 0.29 mM, and 152 to 42 microM and 172 to 49 microM for glycerol concentrations in men and women, respectively (p < 0.05). Isoproterenol-stimulated blood flow also inversely correlated to skin-fold thickness (p < 0.05). We conclude that increased adipose tissue thickness is associated with reduced tissue perfusion and metabolism, even in lean subjects. Skin-fold thickness is an important confounding variable in metabolic studies, particularly in microdialysis experiments. PMID- 15778922 TI - Pyrrolidon carboxypeptidase activities in the hypothalamus-pituitary-thyroid and hypothalamus-pituitary-ovary axes of rats with mammary gland cancer induced by N methyl nitrosourea. AB - Pyrrolidon carboxypeptidase is an omega-peptidase that hydrolyses N-terminal pyroglutamyl residues from biologically active peptides such as gonadotropin releasing and thyrotrophin-releasing hormones. We previously described a decrease in both rat and human pyrrolidon carboxypeptidase activity with breast cancer, suggesting that gonadotropin-releasing hormone may be an important local intracrine, autocrine and/or paracrine hormonal factor in the pathogenesis of breast cancer while playing a role in the tumoral process. However, the other susceptible substrate of pyrrolidon carboxypeptidase, thyrotrophin-releasing hormone, may also be modified with breast cancer, supporting an association between breast cancer and thyroid disorders. The present work analyses soluble and membrane-bound pyrrolidon carboxypeptidase activities in the hypothalamus pituitary-thyroid and hypothalamus-pituitary-ovary axes in N-methyl nitrosourea induced breast cancer in rats. Our aim was to determine the possible relationship between gonadotropin-releasing hormone and thyrotrophin-releasing hormone regulation through pyrrolidon carboxypeptidase activity. We propose that pyrrolidon carboxypeptidase activity dysregulation at various local and systemic levels may participate in the initiation, promotion and progression of breast cancer induced in rat by N-methyl nitrosourea through the increase in gonadotropin-releasing hormone. Since pyrrolidon carboxypeptidase activity also acts on thyrotrophin-releasing hormone, the dysregulation of this enzyme's activity could indirectly affect hypothalamus-pituitary-thyroid axis function, and thus potentially represent a link between the diseases of thyroid and breast cancer. PMID- 15778923 TI - Effect of 5-Campestenone (24-methylcholest-5-en-3-one) on Zucker diabetic fatty rats as a type 2 diabetes mellitus model. AB - We examined the therapeutic effects of dietary exposure to 5-campestenone (24 methylcholest-5-en-3-one), an enone derivative of campesterol, in Zucker diabetic fatty (ZDF) rats, an animal model of type 2 diabetes mellitus. Dietary 0.6 % exposure to 5-campestenone caused marked reduction in hemoglobin A1c (HbA1c), plasma total cholesterol, triglycerides and non esterified fatty acid (NEFA). In particular, plasma triglyceride levels were reduced in the 0.6 % 5-campestenone fed group to about 25 % of that in the control group. In the oral glucose tolerance test (OGTT) at three and seven weeks after the beginning of treatment, 5-campestenone limited the rise of blood glucose levels by oral administration of glucose dose-dependently. Amounts of adipose tissue in the retroperitoneum and periepididymal area as well as abdominal subcutaneous fat were significantly decreased in animals fed 0.6 % 5-campestenone. The blood leptin concentration on the final day of feeding was significantly in animals administered 5 campestenone. No obvious anomaly due to consumption of 5-campestenone was detected in necropsy or clinical observations. PMID- 15778924 TI - Time course of vasopressin and oxytocin secretion after stress in adrenalectomized rats. AB - To characterize the participation of vasopressin (AVP) and oxytocin (OT) in hypothalamus-pituitary-adrenal regulation after adrenalectomy (ADX), we evaluated corticosterone, ACTH, AVP and OT plasma concentrations and AVP and OT content of the paraventricular nucleus (PVN) at different periods (3 h, 1, 3, 7 and 14 days) in sham or ADX rats under basal conditions and after immobilization stress. ADX animals showed undetectable corticosterone levels, while sham animals showed a marked increase in corticosterone and ACTH 3 h after surgery, then lowering to basal control levels. ADX rats showed high basal ACTH levels with a triphasic response without changes after immobilization. After three hours, the ADX group showed higher OT levels than the sham group. OT was increased after immobilization stress in sham and ADX groups. AVP plasma levels did not change throughout the basal or stress studies in either group. There was a decrease in hypothalamic AVP content 1 and 3 days after ADX under basal and stress conditions. Plasma osmolality showed a significant decrease in the ADX group at 3, 7, and 14 days. In conclusion, there are different pituitary-adrenal axis set points after removal of the glucocorticoid negative feedback. The role of vasopressinergic and oxytocinergic neurons in the ACTH secretion after ADX or immobilization stress appears to differ. Magnocellular AVP is unlikely to contribute to ACTH secretion in response to ADX or immobilization stress. On the other hand, OT is elicited by immobilization stress and might contribute to the ACTH secretion during short-term ADX. PMID- 15778925 TI - Anemia under androgen deprivation: influence of flutamide, cyproteroneacetate and orchiectomy on the erythropoietin system. AB - OBJECTIVE: There is a high incidence of anemia in patients with advanced prostate cancer (PC) under androgen deprivation. Pathophysiology of this anemia remains unclear. Erythropoietin (EPO) is the main growth factor inducing erythropoesis in response to hypoxia. In this study, function of the EPO-system following androgen deprivation was tested in standardized animal model. METHODS: Animals were pretreated by either orchiectomy, injection of cyproteroneacetate or flutamide. After hypoxic stimulation, EPO mRNA expression and EPO serum levels were studied. RESULTS: In all animals, EPO mRNA expression and EPO serum levels were increased following hypoxic stimulation. Compared to the control group, this increase was even more pronounced after androgen deprivation. None of the different forms of androgen deprivation had a negative stimulating effect on EPO expression. CONCLUSION: Unexpectedly, androgen deprivation did not suppress EPO mRNA expression and EPO serum concentrations. Instead, stimulation of the EPO system was even more pronounced after androgen deprivation. A deficient EPO system does not appear to contribute to the clinically observed anemia in patients treated by androgen deprivation. PMID- 15778926 TI - Influence of leptin, androgens and insulin sensitivity on increased GH response to clonidine in lean patients with polycystic ovary syndrome. AB - Our aim was to investigate whether insulin sensitivity, leptin, androgen or estradiol levels are associated with disturbed GH response to clonidine in lean patients with polycystic ovary syndrome. Fourteen lean polycystic ovary syndrome patients, 11 ovulatory patients presenting idiopathic hirsutism and 10 non hirsute, normal women with regular cycles paired for age and BMI were included in a cross-sectional study. Baseline hormonal and metabolic variables were assessed and analyzed in association with GH response to oral administration of 0.3 mg of clonidine. Delta GH was significantly higher in the PCOS group than in the IH and control groups (p = 0.014). The groups were similar in terms of body mass index, insulin, glucose, total and HDL cholesterol, triglycerides and estradiol levels. Free androgen index (r = 0. 454, p = 0.015) and leptin (r = 0.419, p = 0.023) were positively correlated with the homeostasis model assessment. The homeostasis model assessment was the only variable that significantly correlated with GH response to clonidine (r = 0.375, p = 0.029) (vs. estradiol, free androgen index, leptin and LH). Nonetheless, when the analysis was adjusted for leptin levels and free androgen index, the statistical significance of this correlation was lost. The increased GH secretion observed in our lean PCOS patients may be associated with slight changes in insulin sensitivity, even in the absence of clinical evidence of insulin resistance. This association seems to be modulated by leptin and androgen levels. PMID- 15778927 TI - Influence of the PPAR-gamma2 Pro12Ala and ACE I/D polymorphisms on insulin sensitivity and training effects in healthy offspring of type 2 diabetic subjects. AB - Aerobic endurance training improves insulin sensitivity, and is of great importance in the prevention and treatment of type 2 diabetes. The improvement in insulin sensitivity and cardiovascular function through exercise is highly variable among individuals, and is probably partly determined by genetic components. This study evaluated the peroxisome proliferation-activated receptor gamma2 ( PPAR-gamma2) Pro12Ala polymorphism and the angiotensin converting enzyme ( ACE) I/D polymorphism with respect to any potential influence that these highly prevalent polymorphisms may impose on changes in insulin sensitivity and maximal aerobic capacity induced by exercise. Seventy-nine healthy first-degree relatives of type 2 diabetic patients were compared to a control group consisting of 54 subjects without any family history of type 2 diabetes. All subjects had a normal OGTT. The groups were comparable with respect to age (34 +/- 7 vs. 33 +/- 7 years), gender ((m/f) 43/36 vs. 30/24) and BMI (25.7 +/- 2.6 vs. 25.3 +/- 2.5 kg/m (2)); p (all) = NS. Furthermore, a subgroup of 29 offspring and 17 control subjects were engaged in a standardized training program lasting ten weeks. Insulin sensitivity (hyperinsulinemic euglycemic clamp technique) and VO (2)max (exhaustive exercise test) was assessed before and after the training period. We will demonstrate the allelic frequency of the Ala-allele of the Pro12Ala polymorphism to be lower in offspring to type 2 diabetic patients (13.3 %) compared to control subjects (21.3 %); p < 0.05. In offspring only, the Pro12Ala polymorphism of the PPAR-gamma2 gene appeared to enhance weight changes brought about by exercise (Deltaweight = - 0.3 +/- 1.4 kg vs. - 1.8 +/- 1.8 kg; p < 0.05; (Pro/Pro vs. Pro/Ala + Ala/Ala) - suggesting possible gene-environment or gene gene interactions. The ACE I/D polymorphism was not of significant importance in determining the capability of responding to exercise in terms of improvement in insulin sensitivity or maximal aerobic capacity. PMID- 15778928 TI - Optimizing insulin sensitivity assessment using the minimal model in chronic heart failure. AB - BACKGROUND: Minimal model analysis of the intravenous glucose tolerance test (IVGTT) has been used successfully to demonstrate that patients with chronic heart failure (CHF) are insulin-resistant. Continuing experience in minimal model methodology has raised questions about how best to assign basal glucose concentrations during such analyses. METHODS AND RESULTS: IVGTT data from randomly selected patients with CHF (n = 15) and controls (n = 15) were analysed using the minimal model, with the basal glucose concentration (G (b)) assigned the value of fasting plasma glucose concentration (G (fast)), or the value of plasma glucose concentration 180 minutes after the start of the IVGTT (G (180)). Insulin sensitivity (S (I)) was significantly higher with G (b) = G (fast), than with G (b) = G (180) (controls: 5.60 +/- 0.78 vs. 3.36 +/- 0.25/min/muU/ml x 10 (4), p = 0.0017; patients 4.19 +/- 0.54 vs. 2.36 +/- 0.15/min/microU/ml x 10 (4), p = 0.0004). At G (b) = G (fast), CHF patients showed a non-significant 25 % reduction in S (I) in comparison to controls (p = 0.15). In contrast, at G (b) = G (180), CHF patients showed a significant 30 % reduction of S (I) in comparison to controls (p = 0.0018). S (I) estimates derived at G (b) = G (fast) exhibited twice the variability of those estimated using G (b) = G (180) (coefficients of variation of S (I) in patients with CHF were 50.0 % and 24.8 %, respectively). CONCLUSION: In studies of patients with CHF, greater precision and discriminatory power of insulin sensitivity estimates is obtained when the basal glucose concentration is taken as the plasma glucose concentration 180 minutes after the start of the IVGTT. PMID- 15778929 TI - Nutrient-stimulated GLP-1 release in normal-weight men and women. AB - The aim of the present study was to investigate the effect of a monosaccharide/fiber preload (galactose/guar gum) in combination with a standard breakfast (GG) on plasma GLP-1, insulin, glucose, free fatty acid concentrations, and appetite ratings (satiety, hunger, fullness and desire to eat) in 30 normal weight subjects. GG was compared to water in combination with the standard breakfast (W). We also tested for differences in GLP-1 release in the described conditions between genders. In women, postprandial plasma GLP-1 concentrations after GG were significantly increased compared to W (p < 0.05). The area under the curve (AUC) for DeltaGLP-1 after GG was related to body fat (%BF) in women (r = 0.48; p = 0.02), but not in men. The rise in plasma insulin was delayed, and plasma glucose concentration was blunted after GG. Deltasatiety was significantly related to DeltaGLP-1 (W). In conclusion, galactose with guar gum increased and extended the GLP-1 release due to breakfast in women, but not in men. This may be partly explained by %BF, which is higher in women than in men since GLP-1 release appears to be related to %BF. PMID- 15778931 TI - Direct effect of leptin on gastric ghrelin secretion. PMID- 15778930 TI - Spontaneous prolactin secretion in growth hormone-deficient children. AB - OBJECTIVE: To establish the spontaneous nocturnal prolactin (PRL) release in relation to growth hormone (GH)-deficient children and idiopathic short-stature children (ISS). METHODS: A total of 32 prepubertal children (11 girls, 21 boys) aged between 3 and 12 years were studied retrospectively and sorted according to diagnosis: idiopathic GH deficiency (GHD, n = 9), neurosecretory deficiency of GH secretion (NSD, n = 10) and ISS (n = 13). Nocturnal spontaneous hormone secretion was studied by intermittent venous sampling. Secretion profiles and copulsatility were analyzed using Pulsar and AnCoPuls software. RESULTS: (median, range in mug/l): Children with GHD and NSD had significantly lower GH and area-under-the curve (AUC) levels than normal children (p < 0.001), whereas ISS children showed normal values. In contrast, prolactin levels were significantly higher (p < 0. 05) in children with GHD and NSD (11.1, 4.9 - 13.0 and 10.3, 8. 8 - 19. 6, respectively) compared to the ISS children (8.0, 4.9 - 13.0). In addition, prolactin AUC and peak height were higher (p < 0.05) in GH-deficient patients, whereas all other secretion parameters were the same. Correlation and copulsatility analysis revealed no evidence for a direct relation between PRL and GH secretion. CONCLUSIONS: PRL secretion is significantly higher in children with GHD and NSD compared to ISS children but PRL and GH show no copulsatile secretion pattern. PMID- 15778933 TI - Cross-tolerance in the tidepool sculpin: the role of heat shock proteins. AB - Cross-tolerance, or the ability of one stressor to transiently increase tolerance to a second heterologous stressor, is thought to involve the induction of heat shock proteins (Hsp). We thus investigated the boundaries of cross-tolerance in tidepool sculpins (Oligocottus maculosus) and their relationship to Hsp70 levels. Survival of sculpins exposed to severe osmotic (90 ppt, 2 h) and hypoxic (0.33 mg O(2)/L, 2 h) stressors increased from 68% to 96%, and from 47% to 76%, respectively, following a +12 degrees C heat shock. The magnitude of this heat shock was critical for protection. A +10 degrees C heat shock did not confer cross-tolerance, while a +15 degrees C heat shock was deleterious. Sculpins required between 8 and 48 h of recovery following the +12 degrees C heat shock to develop cross-tolerance. There was no association between Hsp70 levels before the onset of the secondary stressor and cross-tolerance. However, branchial Hsp70 levels following osmotic shock were highly correlated with the time frame of cross-tolerance. Thus, Hsp70 induction by the priming stressor may be less important than the ability of the cell to mount an Hsp response to subsequent stressors. The time frame of cross-tolerance is similar to the interval between low tides, suggesting the possible relevance of this response in nature. PMID- 15778934 TI - Phenotypic flexibility at the molecular and organismal level allows desert dwelling rodents to cope with seasonal water availability. AB - We examined the phenotypic flexibility of field urine osmolality (Uosm) in response to seasonal rainfall and the experimental expression of renal aquaporins (AQPs) in the leaf-eared mouse Phyllotis darwini, a South American desert dwelling rodent, through an integrative study at both the cellular and the organismal level. Field Uosm was higher in summer than in winter. Fall and winter Uosm were not significantly different. During a rainy year, winter Uosm was 2,140 +/- 82.3 mOsm kg(-1); the corresponding value in a dry year was 2,569 +/- 61.3 mOsm kg(-1). During the summer, the mean Uosm in a rainy year was 3,321 +/- 71.5 mOsm kg(-1), and in a dry year it was 3,604 +/- 107.2 mOsm kg(-1). The distribution of AQP-2, AQP-3, and AQP-4 was similar to that described for mouse and rat kidneys and confined to principal cells in cortex and inner medullary collecting-duct cells. AQP-4 immunoreactivity was unaltered by the state of water balance. Relative to water loading, dehydration induced an increase in AQP-2 immunoreactivity and protein abundance. Although more discrete, AQP-3 immunolabeling was also increased by dehydration. We now reveal how the integration of flexible renal mechanisms acting at the cellular and organismal level allow a small desert-dwelling mammal to cope with seasonal and yearly (El Nino) water availability in its semiarid habitat. PMID- 15778935 TI - The flight physiology of reproductives of Africanized, European, and hybrid honeybees (Apis mellifera). AB - Neotropical African honeybees (Apis mellifera scutellata), in the process of spreading throughout tropical and subtropical regions of the Americas, hybridize with and mostly replace European honeybees (primarily Apis mellifera mellifera and Apis mellifera ligustica). To help understand this process, we studied the effect of lineage (African, European, or hybrid) on the flight physiology of honeybee reproductives. Flight metabolic rates were higher in queens and drones of African lineage than in European or hybrid bees, as has been previously found for foraging workers. These differences were associated with higher thorax/body mass ratios and higher thorax-specific metabolic rates in African lineage bees. Queens were reared in common colonies, so these metabolic and morphological differences are likely to be genetic in origin. African drones had higher wing beat frequencies and thorax temperatures than European or hybrid bees. Hybrids were intermediate for many parameters, but hybrid queen mass-specific flight metabolic rates were low relative to Africans and were nonlinearly affected by the proportion of African lineage, consistent with some negative heterosis for this trait. PMID- 15778936 TI - Intermittent locomotor activity that increases endurance also increases metabolic costs in the desert Iguana (Dipsosaurus dorsalis). AB - Intermittent activity, alternating bouts of activity and rest, can extend endurance relative to continuous locomotion. Utilizing a rapid fatiguing activity intensity (1.08 m s(-1)), Dipsosaurus dorsalis (n = 14) ran repeated bouts of varying durations (5, 15, or 30 s) interspersed with variable pause periods (100%, 200%, 400%, or 800% of the activity period) until exhausted. Total distance ran increased relative to continuous locomotion. The largest increases were seen when activity periods were limited to 5 s and pause periods were extended from 5 s to 20 s to 40 s (55, 118, and 193 m, respectively). To analyze these increases further, O(2) consumption was measured for six bouts of 5-s activity separated by either 5, 20, or 40 s (n = 8). The sum of elevated O(2) consumption during activity, pauses, and recovery increased significantly from 0.08 to 0.09 and 0.12 mL O(2) g(-1) as pause duration increased, primarily due to greater O(2) consumption during longer pause intervals. Postexercise recovery metabolism was a large cost (>57% of total) but did not differ among treatments. Overall, 40-s pauses were most expensive (absolutely and per unit distance) but provided the greatest endurance, likely due to further repletion of metabolites or removal of end products during the longer pause. In contrast, the shortest pause period was most economical but exhausted the animal most rapidly. Thus, a pattern of intermittent activity utilized by an animal may have energetic advantages that sometimes may be offset by behavioral costs associated with fatigue. PMID- 15778937 TI - Stickleback fights: why do winners win? Influence of metabolic and morphometric parameters. AB - Pairs of reproductively mature male three-spined stickleback (Gasterosteus aculeatus) were introduced into unfamiliar aquaria and observed until one male became dominant. Skin carotenoid content, morphometric indexes, and metabolic capacities of the axial and pectoral muscles were examined to establish whether morphological or physiological parameters differentiated winners and losers. Stickleback that initiated fights typically won. Quick initiation led to quick victory. Overall, winners and losers differed in few morphological or metabolic characteristics, but these properties and the differences between these attributes for losers and winners of specific fights were linked with initiation time and fight duration. Morphometric indexes of losers were the primary determinants of initiation time and fight duration, whereas for winners muscle metabolic capacities were linked to these fight characteristics. The greater the hepatosomatic index (HSI) of losers, the longer the fight initiation times. Similarly, losers with high HSI and carotenoid levels resisted defeat longer. In winners, initiation time decreased as axial muscle phosphofructokinase levels increased and citrate synthase levels decreased, whereas the metabolic capacities of the pectoral muscle were linked with time to achieve victory. When losers had greater HSI values than the winners of a specific fight, fight initiation was delayed and fights lasted longer. When losers had higher carotenoid levels than winners, fights also lasted longer. On the other hand, when losers had more visceral fat (fat body mass over somatic mass) than winners, both initiation time and combat duration were reduced. These results suggest that male stickleback assess their physiological status and that of their opponents, in particular the HSI, and adjust their combat strategies accordingly. PMID- 15778938 TI - The effect of meal composition on specific dynamic action in burmese pythons (Python molurus). AB - We quantified the specific dynamic action (SDA) resulting from the ingestion of various meal types in Burmese pythons (Python molurus) at 30 degrees C. Each snake was fed a series of experimental meals consisting of amino acid mixtures, simple proteins, simple or complex carbohydrates, or lipids as well as meals of whole animal tissue (chicken breast, beef suet, and mouse). Rates of oxygen consumption were measured for approximately 4 d after feeding, and the increment above standard metabolic rate was determined and compared to energy content of the meals. While food type (protein, carbohydrate, and lipid) had a general influence, SDA was highly dependent on meal composition (i.e., amino acid composition and carbohydrate structure). For chicken breast and simple carbohydrates, the SDA coefficient was approximately one-third the energetic content of the meal. Lard, suet, cellulose, and starch were not digested and did not produce measurable SDA. We conclude that the cost of de novo protein synthesis is an important component of SDA after ingestion of protein meals because (1) simple proteins, such as gelatin and collagen, did not stimulate levels of SDA attained after consumption of complete protein, (2) incomplete mixtures of amino acids failed to elicit the SDA of a complete mixture, and (3) the inhibition of de novo protein synthesis with the drug cycloheximide caused a more than 70% decrease in SDA. Stomach distension and mechanical digestion of intact prey did not cause measurable SDA. PMID- 15778939 TI - Ultraviolet exposure and vitamin D synthesis in a sun-dwelling and a shade dwelling species of Anolis: are there adaptations for lower ultraviolet B and dietary vitamin D3 availability in the shade? AB - We compared the natural ultraviolet B (UV-B) exposure, dietary vitamin D, and skin-generated vitamin D synthesis for adult males of two species of Jamaican anoles. The more shade-tolerant and thermal-conforming Anolis lineotopus merope, rarely exposed to full sun, experienced less UV-B irradiation in its shady environment than the more heliophilic and thermophilic Anolis sagrei, which frequently basked in full sun during the morning hours (0800-1100 hours). Both species obtained detectable levels of vitamin D(3) in their diet, but the heliophilic A. sagrei obtained more. To compensate for less availability of UV-B and dietary vitamin D, the skin of A. lineotopus merope seems to have acquired a greater sensitivity than that of A. sagrei regarding UV-B-induced vitamin D(3) photobiosynthesis. We assessed this by observing a greater conversion of provitamin D to photoproducts in skin exposed to UV-B from a sunlamp. The reduced skin sensitivity of A. sagrei regarding vitamin D photobiosynthesis may reflect a correlated response associated with less need for vitamin D photobiosynthesis and greater need for UV-B screening capacity as an adaptation to a more damaging UV-B environment. However, the possibility that adaptations for photobiosynthesis of vitamin D and for protection from skin damage could involve independent mechanisms needs investigation. Also, the ability to behaviorally regulate UV-B exposure, as shown for the panther chameleon, would benefit both species of Anolis and should be investigated. PMID- 15778940 TI - Symbiotic fermentation, digesta passage, and gastrointestinal morphology in bullfrog tadpoles (Rana catesbeiana). AB - Relative to other herbivorous vertebrates, the nutritional ecology and digestive physiology of anuran larvae remain poorly understood. Our objective was to compare gut structure and inhabitants, digesta passage, and microbial fermentation in bullfrog tadpoles (Rana catesbeiana) to those in other herbivores. Bullfrog tadpole gastrointestinal tracts were long and voluminous, with an enlarged colon that harbored a diverse symbiotic community. The transit time for particulate markers passing through bullfrog tadpoles was 6 h, the median retention time was 8-10 h, and gut clearance was 10-14 h postingestion. Relatively high levels of short-chain fatty acids in the hindgut of tadpoles indicated active microbial fermentation in this gut region. This report represents the first account of gastrointestinal fermentation in the class Amphibia. On the basis of in vitro fermentation assays, we estimated that microbial fermentation in the hindgut provides 20% of the total daily energy requirement of bullfrog tadpoles. These tadpoles also exhibited coprophagy, a practice that provides important nutritive gains in other herbivores. The physiological and behavioral characteristics of these tadpoles are remarkably similar to those of other small-bodied, hindgut-fermenting vertebrates, suggesting convergent digestive strategies among a broad range of herbivorous taxa. PMID- 15778941 TI - The little skate Raja erinacea exhibits an extrahepatic ornithine urea cycle in the muscle and modulates nitrogen metabolism during low-salinity challenge. AB - Urea synthesis via the hepatic ornithine urea cycle (OUC) has been well described in elasmobranchs, but it is unknown whether OUC enzymes are also present in extrahepatic tissues. Muscle and liver urea, trimethylamine oxide (TMAO), and other organic osmolytes, as well as selected OUC enzymes (carbamoyl phosphate synthetase III, ornithine transcarbamoylase, arginase, and the accessory enzyme glutamine synthetase), were measured in adult little skates (Raja erinacea) exposed to 100% or 75% seawater for 5 d. Activities of all four OUC enzymes were detected in the muscle. There were no changes in muscle OUC activities in skates exposed to 75% seawater; however, arginase activity was significantly lower in the liver, compared to controls. Urea, TMAO, and several other osmolytes were significantly lower in the muscle of little skates exposed to 75% seawater, whereas only glycerophosphorylcholine was significantly lower in the liver. Urea excretion rates were twofold higher in skates exposed to 75% seawater. Taken together, these data suggest that a functional OUC may be present in the skeletal muscle tissues of R. erinacea. As well, enhanced urea excretion rates and the downregulation of the anchor OUC enzyme, arginase, in the liver may be critical in regulating tissue urea content under dilute-seawater stress. PMID- 15778942 TI - Temperature influences whole-animal rates of metabolism but not protein synthesis in a temperate intertidal isopod. AB - The effects of temperature on two important biological rate processes, whole-body rates of oxygen uptake (M dot o2) and protein synthesis (k(s)), were investigated in the temperate intertidal isopod Ligia oceanica at two different times of the year. Animals were collected in January (winter) and June (summer) and either subjected to an acute temperature change after 24 h (acclimatized) or acclimated to various temperatures for 4 wk. In both cases, M dot o2 increased with temperature, with a Q(10) of 2.2 between 5 degrees and 20 degrees C, but increased in thermal sensitivity at 25 degrees C. Winter isopods were characterized by significantly higher M dot o2 levels, greater thermal sensitivities, and lower thermal tolerances than summer animals. Seasonal differences in M dot o2 persisted after acclimation, indicating that temperature alone was not responsible for the changes. In sharp contrast, whole-body k(s) showed no variation with temperature, although overall rates decreased upon acclimation. In acclimatized animals, k(s) was higher in the summer than in the winter. After acclimation, a compensatory increase in RNA capacity in winter animals reversed this situation. The temperature independence of whole-body k(s) in L. oceanica could ensure survival in a highly liable thermal environment, as thermal tolerances of intertidal invertebrates are thought to be more closely related to protein than to energy metabolism. PMID- 15778943 TI - Are the low protein requirements of nectarivorous birds the consequence of their sugary and watery diet? A test with an omnivore. AB - Nectar-feeding birds have remarkably low nitrogen requirements. These may be due either to adaptation to a low-protein diet or simply to feeding on a fluid diet that minimizes metabolic fecal nitrogen losses. We measured minimal nitrogen requirements (MNR) and total endogenous nitrogen loss (TENL) in the omnivorous European starling Sturnus vulgaris, fed on an artificial nectar-like fluid diet of varying concentrations of sugar and protein. The MNR and TENL of the birds were similar and even slightly higher than allometrically expected values for birds of the starlings' mass (140% and 103%, respectively). This suggests that the low measured nitrogen requirements of nectar-feeding birds are not simply the result of their sugary and watery diets but a physiological adaptation to the low nitrogen input. We also measured the effect of water and protein intake on the nitrogenous waste form in the excreta and ureteral urine in European starlings. Neither high water intake nor low protein intake increased the fraction of nitrogen excreted as ammonia. Ammonia was excreted at consistently low levels by the starlings, and its concentration was significantly higher in ureteral urine than in excreta. We hypothesize that ureteral ammonia was reabsorbed in the lower intestine, indicating a postrenal modification of the urine. PMID- 15778944 TI - Experimental evidence of food-independent larval development in endemic Jamaican freshwater-breeding crabs. AB - In an experimental study, we compared reproductive and developmental traits of endemic sesarmid crabs from Jamaica living in landlocked limnic or terrestrial habitats. Laboratory rearing and behavioral observations showed that the larval development of Sesarma windsor, Sesarma dolphinum (both from freshwater brooks), and Metopaulias depressus (the bromeliad crab) invariably consists of two nonfeeding zoeal stages and a facultatively lecithotrophic megalopa. In a quantitative study of life-history processes characterizing this developmental mode, we provide for S. windsor first data of biomass and elemental composition (dry mass, W; carbon, hydrogen, nitrogen, collectively, CHN) during development from the egg through successive larval stages. These data show that larval independence of food is based on an enhanced female energy allocation in reproduction, reflected in unusually large egg size (1.45-1.70 mm), as well as high contents of C and H (about 60% and 9%, respectively) and high C : N ratios (7.6-8.4) in eggs and early larvae. During zoeal development, about 6% of initial W and 9% of N but 13% each of C and H were lost; similar losses occurred during megalopal development in continued absence of food. These patterns reflect the metabolic utilization of stored organic matter, with preferential degradation of lipid reserves. Fed megalopae gained greater amounts of W and N as compared with C and H (increments of 37% and 38% vs. 25% and 19%, respectively), indicating preferential investment of nutritional energy in proteins required for the formation of new tissues and organs, while generally decreasing proportions of CHN within total W suggested an increasing mineralization of the exoskeleton. Although survival and molt cycle duration of the megalopa stage were not affected by absence of food, significant effects were found in the size of first-stage juvenile crabs, indicating a trade-off between nutritional flexibility in the last larval stage and postmetamorphic fitness. Similar patterns of development and biomass in M. depressus as well as preliminary data obtained for S. dolphinum and Sesarma fossarum suggest that reproductive and developmental traits may be similar in all endemic Jamaican sesarmids. These traits are interpreted as life history adaptations to development in landlocked habitats, probably playing a key role during adaptive radiation. PMID- 15778946 TI - Breathing hypoxic gas affects the physiology as well as the diving behaviour of tufted ducks. AB - We measured the effects of exposure to hypoxia (15% and 11% oxygen) and hypercapnia (up to 4.5% carbon dioxide) on rates of respiratory gas exchange both between and during dives in tufted ducks, Aythya fuligula, to investigate to what extent these may explain changes in diving behaviour. As found in previous studies, the ducks decreased dive duration (t(d)) and increased surface duration when diving from a hypoxic or hypercapnic gas mix. In the hypercapnic conditions, oxygen consumption during the dive cycle was not affected. Oxygen uptake between dives was reduced by only 17% when breathing a hypoxic gas mix of 11% oxygen. However, estimates of the rate of oxygen metabolism during the foraging periods of dives decreased nearly threefold in 11% oxygen. Given that tufted ducks normally dive well within their aerobic dive limits and that they significantly reduced their t(d) during hypoxia, it is not at all clear why they make this physiological adjustment. PMID- 15778945 TI - A pharmacological examination of Na+ and Cl- transport in two species of freshwater fish. AB - We examined branchial Na(+) and Cl(-) uptake in two species of stenohaline, freshwater fish (goldfish and the Amazonian neon tetra). Kinetic analysis revealed that the two species had similar uptake capacities and affinities for Na(+) and Cl(-). However, while uptakes of Na(+) and Cl(-) (JNain and JClin, respectively) by goldfish were completely inhibited at pH 4.5 and below, uptake in tetras was unaffected by pH down to 3.25. Examination of Cl(-) transport with blockers indicated that goldfish and neon tetras utilize Cl(-)/HCO-3 exchange; SITS and SCN(-) inhibited Cl(-) uptake in both species. In contrast, large differences in Na(+) transport were indicated between the species. In goldfish, exposure to four Na(+)/H(+) exchange blockers, as well as the Na(+) channel blocker phenamil, strongly inhibited JNain. Further, Na(+) and Cl(-) uptake were strongly inhibited by the Na(+)/K(+)/Cl(-) cotransport inhibitor furosemide, as was JNain in "Cl(-)-free" water and JClin in "Na(+)-free" water. This suggests the presence of multiple transporters and possibly even a direct linkage between the transport of Na(+) and Cl(-) in goldfish. In contrast, none of these drugs strongly reduced Na(+) transport in neon tetras, which raises the possibility of a significantly different Na(+) transport mechanism in this acid-tolerant species. PMID- 15778947 TI - Effects of diving and swimming behavior on body temperatures of pacific leatherback turtles in tropical seas. AB - Mathematical models and recordings of cloacal temperature suggest that leatherback turtles (Dermochelys coriacea) maintain core body temperature higher than ambient water temperature (T(W)) while freely swimming at sea. We investigated the thermoregulatory capabilities of free-ranging leatherbacks and, specifically, the effect that changes in diving patterns and ambient temperatures have on leatherback body temperatures (T(B)). Data loggers were used to record subcarapace and gastrointestinal tract temperatures (T(SC) and T(GT), respectively), T(W), swim speed, dive depth, and dive times of female leatherback turtles during internesting intervals off the coast of Guanacaste, Costa Rica. Mean T(SC) (28.7 degrees -29.0 degrees C) was significantly higher than mean T(W) (25.0 degrees -27.5 degrees C). There was a significant positive relationship between T(SC) and T(W) and a significant negative correlation between T(SC) and dive depth and T(GT) and dive depth. Rapid fluctuations in T(GT) occurred during the first several days of the internesting interval, which suggests that turtles were ingesting prey or water during this time. Turtles spent 79%-91% of the time at sea swimming at speeds greater than 0.2 m s(-1), and the average swim speed was 0.7 +/- 0.2 m s(-1). Results from this study show that alterations in diving behavior and T(W) affect T(B) of leatherback turtles in the tropics. Body temperatures of free-ranging leatherback turtles correspond well with values for T(B) predicted by mathematical models for tropical conditions. PMID- 15778948 TI - Determination of HIV-1 subtypes (A-D, F, G, CRF01_AE) by PCR in the transmembrane region (gp41) with novel primers. AB - HIV-1 has a huge genetic diversity. So far, nine subtypes have been isolated, namely, subtypes A, B, C, D, F, G, H, J, and K. Epidemiological study provides information which may help in the development of HIV-1 prevention programs or health policies. In the future, subtyping may also be critical for vaccine development, and an effective anti-viral drug will need to be effective for different subtypes of HIV virus. The analysis of the nucleotide sequence of the v3 region is considered the most reliable method for determining the HIV-1 subtype. However, the procedures for determining the v3 sequences are complicated and time consuming, requiring expensive reagents, equipment, and well-trained personnel. The polymerase chain reaction (PCR) method using subtype-specific primers for HIV-1 subtyping is easier and faster. The objective of this study was to develop subtype-specific primers for subtyping PCR. The specific primers were designed for subtypes A, B, C, D, F, G, and CRF01_AE, and these primers could be applied to assay for various HIV-1 subtypes in the clinical samples. The specific primers were designed for each subtypes in the gp41 region. The result of PCR was compared with the subtypes which was determined by the v3 sequence. The results of subtyping by PCR using the newly designed primers could detect 29 of 33 patients tested, and all matched those obtained by nucleotide sequencing of the env v3 region except for three subjects, which were differentiated as CRF02_AG. The newly designed primers functioned accurately and conclusively. In comparison with PCR as a method for the determination of subtypes, sequence analysis requires better-trained personnel, more expensive reagents, and more equipment and time. PMID- 15778949 TI - Subcortical lesions after transient thread occlusion in the rat: T2-weighted magnetic resonance imaging findings without corresponding sensorimotor deficits. AB - PURPOSE: To investigate infarct evolution and functional consequences of exclusive subcortical or cortico-subcortical strokes, transient middle cerebral artery occlusion (MCAO) was conducted in Wistar rats. MATERIALS AND METHODS: MCAO was induced in male Wistar rats (260-300 g) for 60 minutes. Lesion volumes and absolute T2 times on magnetic resonance imaging (MRI) were assessed 1 and 14 days after MCAO using a 4.7-T MRI animal scanner in conjunction with functional testing (adhesive tape removal, cylinder test, and ledged beam walking). RESULTS: Functional test scores were not distinguishable between sham-operated animals (N = 5) and those with exclusive caudoputaminal infarct (N = 8; group cp), but showed significant deficits in animals with cortico-subcortical infarction (N = 10; group cp+). The cp group had lower absolute T2 times and a more pronounced reduction in T2 lesion volume over time than the subcortical component in the cp+ group. There was no correlation of T2 lesion size or absolute T2 times and functional impairment in either group. CONCLUSION: When judged from functional tests alone, subcortical ischemic lesions may not be diagnosed reliably. Furthermore, T2-weighted (T2-w) MRI does not well anticipate functional deficits in primarily striatal lesions. PMID- 15778950 TI - Myocardial blood flow in patients with dilated cardiomyopathy: quantitative assessment with velocity-encoded cine magnetic resonance imaging of the coronary sinus. AB - PURPOSE: To quantify global myocardial perfusion using magnetic resonance imaging (MRI) in patients with heart failure due to idiopathic dilated cardiomyopathy (IDC) and to compare myocardial perfusion and microvascular reactivity with healthy subjects. MATERIALS AND METHODS: A total of 19 subjects (healthy volunteers (N = 12) and IDC patients (N = 7)) were studied using cine MRI to measure left ventricular (LV) mass and a velocity-encoded cine MRI technique to measure coronary sinus flow at rest and after dipyridamole-induced hyperemia. Absolute values of total myocardial blood flow (MBF) were calculated from coronary sinus flow and LV mass. RESULTS: At baseline, MBF was not significantly different in patients with IDC (0.48 +/- 0.07 mL/minute/g) and healthy subjects (0.55 +/- 0.19 mL/minute/g, P= 0.41). After dipyridamole administration, MBF in IDC patients increased to a level significantly less than that in normal volunteers (1.05 +/- 0.35 mL/minute/g vs. 1.99 +/- 1.05 mL/minute/g, P < 0.05). Consequently, MBF reserve was impaired in patients with IDC (2.19 +/- 0.77) compared to that in healthy subjects (3.51 +/- 1.29, P < 0.05). A moderate correlation was found between MBF reserve and LV ejection fraction (r = 0.48, P < 0.05). CONCLUSION: MBF reserve is reduced in patients with IDC, indicating that coronary microcirculatory flow is impaired. This integrated MRI approach allows quantitative measurement of global MBF in humans and may have the potential to study the effects of pharmacological interventions on myocardial perfusion. PMID- 15778951 TI - Conspicuity of liver hemangiomas: short tau inversion recovery, T1, and T2 imaging with gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid. AB - PURPOSE: To compare conspicuity of liver hemangiomas on STIR, T1-weighted, and T2 weighted magnetic resonance (MR) images before and after administration of gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid (Gd-EOB-DTPA) (hepatocellular contrast agent), using contrast-to-noise ratios (CNRs). MATERIALS AND METHODS: Thirteen hemangiomas were imaged using breath-hold gradient echo (GRE) T1, fat-saturated turbo spin echo (TSE)-T2, and short tau inversion recovery (STIR) sequences. Background noise and signal-to-noise ratios (SNRs) for liver and hemangioma, along with CNR for normal liver and hemangioma, were measured on each sequence before and after administration of Gd-EOB-DTPA. Hemangioma conspicuity was also evaluated qualitatively. RESULTS: After Gd-EOB DTPA administration, the quantitative liver SNR decreased 54% on STIR, increased 45% on T1-weighted images, and increased 14.5% on TSE-T2-weighted images. The CNR for liver and hemangioma increased 50% on STIR images (P < 0.0001), increased 46% on T1-weighted imaging (P = 0.0033), and increased 22% on TSE-T2-weighted MR imaging (MRI) (P = 0.0083). After contrast, the CNR for TSE-T2 images was greater than those for both the T1 and STIR images (P < 0.0001 for both). Qualitatively, signal change was visually apparent in the liver on T1 and STIR, but not on T2 images or in the hemangiomas on any sequence. CONCLUSION: Despite the statistically significant T1 and STIR increase in CNR, liver hemangiomas were most conspicuous on TSE-T2 images after Gd-EOB-DTPA. This pilot study with hemangiomas highlights the newly recognized potential benefit of TSE-T2 imaging with hepatocellular contrast. PMID- 15778952 TI - Dynamic Gd-DTPA enhanced MRI as a surrogate marker of angiogenesis in tissue engineered bladder constructs: a feasibility study in rabbits. AB - PURPOSE: To evaluate the potential of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to assess angiogenesis in tissue-engineered bladder constructs in a blinded animal study, and compare different analysis approaches and their correlation with microvessel density (MVD). MATERIALS AND METHODS: Constructs fortified with vascular endothelial growth factor (VEGF) for enhanced vascularity were grafted onto the bladder in nine rabbits. DCE-MRI of Gd-DTPA uptake was performed and analyzed using Tofts' model, the area under the concentration time curve (AUC), and the uptake slope. DCE-MRI parameters were compared to MVD determined with CD31 immunohistochemistry. RESULTS: Significantly increased MVD was found in the high VEGF group (20 ng/g of tissue) but not at low VEGF (10 ng/g) (2.3x increase, P = 0.035 vs. 1.1x over control). Enhanced permeability at low VEGF was suggested by elevated K(trans), but overall correlation to MVD was poor. Significant correlation to MVD was obtained with AUC(8min) (r = 0.705, P = 0.034). Furthermore, AUC(8min) provided the most precise discrimination between different VEGF preparations and was the only parameter to show a significant increase (P = 0.0058) consistent with MVD changes at high VEGF. CONCLUSION: Findings support DCE-MRI for evaluating angiogenesis in bladder constructs and suggest vessel changes other than density. Future studies should incorporate larger contrast agents and permeability assessment to devise an optimal DCE-MRI strategy. PMID- 15778953 TI - Contrast-enhanced magnetic resonance angiography of the lower extremities: standard-dose vs. high-dose gadodiamide injection. AB - PURPOSE: To compare the efficacy and safety of two different doses (0.1 and 0.3 mmol/kg of body weight [BW]) of gadodiamide for contrast-enhanced magnetic resonance angiography (ce-MRA) of the lower extremities with intraarterial digital subtraction angiography (IA-DSA). MATERIALS AND METHODS: A total of 30 patients with peripheral arterial occlusive disease underwent IA-DSA and ce-MRA from the aortic bifurcation down to the ankle. Patients were randomized to receive a total dose of 0.1 or 0.3 mmol/kg of BW gadodiamide (Omniscan, Amersham Buchler), administered intravenously as a series of three automatic bolus injections. Ce-MRA was performed with a 1.5-T system using a body phased-array coil, centered stepwise over the calf, thigh, and pelvic region. A fast T1 weighted, three-dimensional gradient-echo sequence was obtained before and after injection of the allocated dose. IA-DSA was performed using the Seldinger technique and a femoral approach. The vessels under investigation were divided into 31 segments, and ce-MRA and IA-DSA image sets were evaluated in a double blind fashion for the presence of stenosis, presence of collateral vessels, vessel delineation, and overall image quality. Both dose groups were compared with regard to contrast index (CI) and signal- and contrast-to-noise ratios (SNR, CNR). The occurrence of adverse events or side effects was also documented. Sensitivity, specificity, and accuracy were calculated in relation to the results of stenosis grading. RESULTS: A total of 26 patients were entered in the efficacy evaluation, while all 30 patients were included in the safety assessment. The sensitivity, specificity, and accuracy for the 0.1 and 0.3 mmol/kg dose groups were 78.8%/93.0%/88.9% vs. 60.2.%/91.5%/83.2%, respectively. The detection of collaterals was similar to IA-DSA for the 0.3 mmol/kg dose group (30.2% vs. 27.4%), but was lower in the 0.1 mmol/kg dose group (27.3% vs. 12.3%). The high dose gadodiamide injection proved to be superior to the 0.1 mmol/kg dose group with regard to vessel delineation and overall image quality (P = 0.007 and P = 0.002, respectively). The difference between the two dose groups regarding CI, SNR, and CNR was significant (P = 0.0001), in favor of the 0.3 mmol/kg dose group. No adverse events were observed in any of the patients. CONCLUSION: Ce-MRA with gadodiamide is safe and efficacious. Comparison of two different doses with IA-DSA as the standard of reference showed that the 0.3 mmol/kg dose is superior to the standard 0.1 mmol/kg dose with respect to contrast enhancement, vessel delineation, image quality, and detection of collaterals. However, the 0.1 mmol/kg dose was superior to the high dose in the grading of stenosis. PMID- 15778954 TI - Standardized assessment of whole body adipose tissue topography by MRI. AB - PURPOSE: To assess standardized whole body adipose tissue topography in a cohort of subjects at an increased risk for type 2 diabetes and to compare fat distribution in subgroups regarding anthropometric (age, body mass index [BMI]) and metabolic parameters (insulin sensitivity). MATERIALS AND METHODS: A total of 80 volunteers (40 females, 40 males) underwent T1-weighted MR imaging of the entire body. Standardized adipose tissue (AT) profiles were calculated considering the different body structure of the participants. The measured data were interpolated to a defined number of sampling points enabling a direct comparison of the profiles independent on body structure. Resulting mean profiles and region-dependent standard deviations of four age groups and three BMI-groups were compared for females and males. Correlations between insulin sensitivity and body fat distribution were analyzed. RESULTS: Reliable adipose tissue profiles could be obtained from all volunteers. In BMI-matched subgroups, females show significant higher AT and subcutaneous abdominal AT (P < 0.05 both), but lower visceral AT (P < 0.01) compared to the males. Furthermore, visceral AT increases with age, as shown in the matched age groups. In both gender groups, insulin resistant subjects are characterized by higher visceral adipose tissue (VAT) compared to insulin-sensitive subjects. In addition, profiles of insulin resistant subjects show more AT in the shoulder/neck region but less AT in the upper extremities. CONCLUSION: Standardized assessment of whole body AT profiles based on T1-weighted MRI provides a reliable basis for interindividual comparison of the body fat distribution and allows a fast and reliable quantification of total body adipose tissue and the distribution of different AT components as subcutaneous and visceral fat in different body regions. Differences in standardized profiles might enable an early identification of people at risk of metabolic disorders, as not only the amount but also the distribution of AT is expected to play an essential role in the pathogenesis of metabolic diseases. PMID- 15778955 TI - Induced refolding of a temperature denatured llama heavy-chain antibody fragment by its antigen. AB - In a previous study we have shown that llama VHH antibody fragments are able to bind their antigen after a heat shock of 90 degrees C, in contrast to the murine monoclonal antibodies. However, the molecular mechanism by which antibody:antigen interaction occurs under these extreme conditions remains unclear. To examine in more detail the structural and thermodynamic aspects of the binding mechanism, an extensive CD, ITC, and NMR study was initiated. In this study the interaction between the llama VHH -R2 fragment and its antigen, the dye Reactive Red-6 (RR6) has been explored. The data show clearly that most of the VHH-R2 population at 80 degrees C is in an unfolded conformation. In contrast, CD spectra representing the complex between VHH-R2 and the dye remained the same up to 80 degrees C. Interestingly, addition of the dye to the denatured VHH-R2 at 80 degrees C yielded the spectrum of the native complex. These results suggest an induced refolding of denatured VHH-R2 by its antigen under these extreme conditions. This induced refolding showed some similarities with the well established "induced fit" mechanism of antibody-antigen interactions at ambient temperature. However, the main difference with the "induced fit" mechanism is that at the start of the addition of the antigen most of the VHH molecules are in an unfolded conformation. The refolding capability under these extreme conditions and the stable complex formation make VHHs useful in a wide variety of applications. PMID- 15778956 TI - The effect of resolution-dependent global shape modifications on rigid-body protein-protein docking. AB - Docking unbound molecules presents a challenge in the case where no prior biological or bioinformatic knowledge exists. This is mainly due to differences between the structures of the molecules when in a complex and in the free state. Presumably, these differences interfere with the ability of protein-protein docking algorithms, which rely on a dominant shape descriptor, to identify the correct solution and rank it higher than false solutions. In this study we verify the notion that small discords in the molecular fit can be eliminated by using appropriately designed low-resolution shape descriptors, thereby improving the docking results. We exploit the inherent gradual resolution dependency of Fourier transforms and formulate a resolution-dependent shape descriptor by truncating selected Fourier transform terms. Thus, different levels of shape modification are attained, affecting the degree of detail in the depiction of the molecular surface. We applied the modified descriptor to a selection of 23 protein-protein systems, using the unbound structures where possible. The docking results obtained with the new geometric descriptor were considerably superior to former results, improving the ranks of nearly correct solutions for 17 systems. Unification of the results of scans in which different resolutions were employed further improved the ranks of nearly correct solutions to less than 100 for 12 of the 23 systems and less than 300 for 20 systems. The new geometric descriptor can be combined with other descriptors, which typify the electrostatic or hydrophobic character of the molecular surface, and with external experimental or bioinformatic data. PMID- 15778957 TI - Transmission of G145R mutant of HBV to an unrelated contact. AB - Household contacts of HBV-related chronic liver disease patients constitute a high-risk group for acquisition of HBV infection. Some of the HBsAg mutants are associated with liver disease and some are reported to be transmitted vertically. There is limited information on the horizontal transmission of Gly 145 Arg (G145R) mutant to related contacts. Its possible transmission to an unrelated third degree contact is reported in the present study. An HBV related chronic liver disease patient; the index patient, and his 11 household contacts were studied. This included four 1 degrees, three 2 degrees, one 3 degrees, and a sexual contact. Surface gene sequencing including the "a" determinant region was carried out in HBV DNA+ve subjects. The sequences were aligned and compared for the homology. HBV DNA was found to be positive in one 1 degrees, three 2 degrees, and one 3 degrees contact, besides the index patient. Histopathological studies revealed evidence of chronic hepatitis in all these contacts. Mutation T118V was present in all the six subjects. Mutant G145R along with T118V and T143M was identified in three subjects who included one 1 degrees, one 2 degrees, and one 3 degrees contact. Presence of T118V and T143M mutations along with G145R mutation in these subjects provides an indirect evidence for the possible horizontal transmission of G145R HBV variant to a 3 degrees unrelated contact. Of these three contacts with G145R mutation, only one 1 degrees contact was found to be HBsAg-ve. The data also reaffirms the earlier finding of HBsAg positivity in presence of G145R mutation of the S-gene. HBV exists as quasi-species and mixed population in subjects with chronic HBV infection. PMID- 15778958 TI - The incidence of congenital cytomegalovirus infections in The Netherlands. AB - Cytomegalovirus (CMV) is one of the most common causes of congenital infection without an effective treatment or an effective vaccine available to date. The emphasis has to be on preventive strategies, which rely on the epidemiological situation. The incidence of congenital CMV infections, however, is not known for The Netherlands. Therefore, a prospective virological study was carried out in a population of 7,524 pregnant women and 7,793 newborns. CMV-specific IgG antibodies were determined in cord blood by ELISA. When CMV antibodies were present, a CMV specific PCR was performed on the throat swab. A positive PCR was confirmed by urine culture. In addition, the seroepidemiology for CMV was investigated in the metropolitan region (Amsterdam and Rotterdam) which has a different ethnic composition. Congenital CMV infection was found in 7 infants (0.9 per 1,000). None had symptoms at birth or during 24 month follow-up. Carriage or CMV was 41%, with a variation between 35% and 100% depending on ethnicity. The ethnic composition in the south-eastern region was different from that in large cities, but similar to that in the rest of the country. The incidence of congenital CMV infections in The Netherlands is the lowest described to date, which does not justify special preventive policies. PMID- 15778959 TI - Why are rare traits unilaterally expressed?: trait frequency and unilateral expression for cranial nonmetric traits in humans. AB - Based on an analysis of nonmetric trait databases from several large skeletal series in Northern Europe and South America, representing 27 bilateral traits, we report a predictable relationship between the frequency of nonmetric traits and the probability that they are expressed bilaterally. In a wider sampling of traits and populations, this study thus confirms the findings of an earlier study by Ossenberg ([1981] Am. J. Phys. Anthropol. 54:471-479), which reported the same relationship for two mandibular traits. This trend was previously explained by extending the multifactorial threshold model for discontinuous traits to incorporate either separate thresholds for unilateral or bilateral expression, or by a fuzzy threshold in which the probability of bilateral expression increases away from the median threshold value. We show that the trend is produced under the standard multifactorial threshold model for discontinuous traits simply if the within-individual or developmental instability variance remains relatively constant across the range of liability. Under this assumption, the number of individuals in which one side but not the other is pushed over the threshold for trait formation will be a larger proportion of the number of individuals expressing the trait when the trait frequency is low. As trait frequency increases, the significance of within-individual variance as a determinant of trait formation decreases relative to the genetic and among-individual environmental variance. These results have implications for interpreting nonmetric trait data as well as for understanding the prevalence of unilateral vs. bilateral expression of a wide variety of discontinuous traits, including dysmorphologies in humans. PMID- 15778960 TI - Analysis of correlated domain motions in IgG light chain reveals possible mechanisms of immunological signal transduction. AB - It was shown experimentally that binding of a micelle composed of Congo red molecules to immunological complexes leads to the enhanced stability of the latter, and simultaneously prevents binding of a complement molecule (C1q). The dye binds in a cavity created by the removal of N-terminal polypeptide chain, as observed experimentally in a model system-immunoglobulin G (IgG) light chain dimer. Molecular Dynamics (MD) simulations of three forms of IgG light chain dimer, with and without the dye, were performed to investigate the role of N terminal fragment and self-assembled ligand in coupling between V and C domains. Root-mean-square distance (RMSD) time profiles show that removal of N-terminal fragment leads to destabilization of V domain. A micelle composed of four self assembled dye molecules stabilizes and fixes the domain. Analysis of root-mean square fluctuation (RMSF) values and dynamic cross-correlation matrices (DCCM) reveals that removal of N-terminal fragment results in complete decoupling between V and C domains. Binding of self-assembled Congo red molecules improves the coupling, albeit slightly. The disruption of a small beta-sheet composed of N and C-terminal fragments of the domain (NC sheet) is the most likely reason for the decoupling. Self-assembled ligand, bound in the place originally occupied by N-terminal fragment, is not able to take over the function of the beta-sheet. Lack of correlation of motions between residues in V and C domains denotes that light chain-Congo red complexes have hampered ability to transmit conformational changes between domains. This is a likely explanation of the lack of complement binding by immunological complexes, which bind Congo red, and supports the idea that the NC sheet is the key structural fragment taking part in immunological signal transduction. PMID- 15778961 TI - Seasonality and clinical features of human metapneumovirus infection in children in Northern Alberta. AB - Human metapneumovirus (hMPV) causes respiratory tract infections in all age groups. The characteristics of pediatric hMPV infection in Northern Alberta have not been studied. The objectives of this study were to determine the seasonality of pediatric hMPV infections over a 13-month period, the genetic relationship of hMPV isolates to hMPV detected in other parts of Canada, and the burden of illness and possible risk factors for pediatric hMPV hospitalization. Detection of hMPV by polymerase chain reaction was performed on nasopharyngeal specimens collected from outpatients and inpatients at the Stollery Children's Hospital in Edmonton, Alberta, November 12, 2002-December 31, 2003. Forty-two of 1,079 specimens were positive for hMPV (3.9%) from 41 patients (14 outpatients and 27 inpatients), with a peak incidence during January-April, but isolates were detected 10 months of the year. Co-infection was not detected in 39 specimens from which RSV had been detected. Two hMPV genetic clusters were detected, and the isolates were homologous to those of previous Canadian isolates. Four of the 14 outpatients had reactive airways disease. Possible risk factors in the 27 inpatients included prematurity (n = 8), congenital heart disease (n = 6), gastroesophageal reflux disease or aspiration (n = 6), global developmental delay (n = 5), and multiple congenital anomalies (n = 4). Risk factors for hospitalization appear to be similar to risk factors for respiratory syncytial virus hospitalization. PMID- 15778963 TI - Protein secondary structure prediction with dihedral angles. AB - We present DESTRUCT, a new method of protein secondary structure prediction, which achieves a three-state accuracy (Q3) of 79.4% in a cross-validated trial on a nonredundant set of 513 proteins. An iterative set of cascade-correlation neural networks is used to predict both secondary structure and psi dihedral angles, with predicted values enhancing the subsequent iteration. Predictive accuracies of 80.7% and 81.7% are achieved on the CASP4 and CASP5 targets, respectively. Our approach is significantly more accurate than other contemporary methods, due to feedback and a novel combination of structural representations. PMID- 15778962 TI - Characterization of monoclonal antibodies to Marburg virus nucleoprotein (NP) that can be used for NP-capture enzyme-linked immunosorbent assay. AB - After the first documented outbreak of Marburg hemorrhagic fever identified in Europe in 1967, several sporadic cases and an outbreak of Marburg hemorrhagic fever have been reported in Africa. In order to establish a diagnostic system for Marburg hemorrhagic fever by the detection of Marburg virus nucleoprotein, monoclonal antibodies to the recombinant nucleoprotein were produced. Two clones of monoclonal antibodies, MAb2A7 and MAb2H6, were efficacious in the antigen capture enzyme-linked immunosorbent assay (ELISA). At least 40 ng/ml of the recombinant nucleoprotein of Marburg virus was detected by the antigen-capture ELISA format. The epitope of the monoclonal antibody (MAb2A7) was located in the carboxy-terminus of nucleoprotein from amino acid position 634 to 647, while that of the MAb2H6 was located on the extreme region of the carboxy-terminus of the Marburg virus nucleoprotein (amino acid position 643-695). These monoclonal antibodies strongly interacted with the conformational epitopes on the carboxy terminus of the nucleoprotein. Furthermore, these two monoclonal antibodies were reacted with the authentic Marburg virus antigens by indirect immunofluorescence assay. These data suggest that the Marburg virus nucleoprotein-capture ELISA system using the monoclonal antibodies is a promising technique for rapid diagnosis of Marburg hemorrhagic fever. PMID- 15778964 TI - Molecular dynamics simulation of the aggregation of the core-recognition motif of the islet amyloid polypeptide in explicit water. AB - The formation of amyloid fibrils is associated with major human diseases. Nevertheless, the molecular mechanism that directs the nucleation of these fibrils is not fully understood. Here, we used molecular dynamics simulations to study the initial self-assembly stages of the NH2-NFGAIL-COOH peptide, the core recognition motif of the type II diabetes associated islet amyloid polypeptide. The simulations were performed using multiple replicas of the monomers in explicit water, in a confined box starting from a random distribution of the peptides at T = 300 K and T = 340 K. At both temperatures the formation of unique clusters was observed after a few nanoseconds. Structural analysis of the clusters clearly suggested the formation of "flat" ellipsoid-shaped clusters through a preferred locally parallel alignment of the peptides. The unique assembly is facilitated by a preference for an extended conformation of the peptides and by intermolecular aromatic interactions. Taken together, our results may provide a description of the molecular recognition determinants involved in fibril formation, in terms of the atomic detailed structure of nascent aggregates. These observations may yield information on new ways to control this process for either materials development or drug design. PMID- 15778965 TI - Induction of dendritic cell-mediated immune responses against HIV-1 by antigen capturing nanospheres in mice. AB - Prophylactic vaccines, designed to elicit potent humoral and cellular immune responses to human immunodeficiency virus type 1 (HIV-1) antigens in mucosa, are the important approach to the protection of individuals against HIV-1 infection, since HIV-1 transmission is largely a result of sexual contact. In this study, a novel strategy has been developed to induce HIV-1-specific immune responses, which involves inactivated HIV-1-caputring concanavalin A (Con A)-immobilized nanospheres (HIV-NS) and their interaction with bone marrow (BM)-derived dendritic cells. HIV-NS were taken up by dendritic cells via cytoskeleton dependent but mannose-binding site-independent phagocytosis. Serial stimulations to unprimed T-cells with HIV-1 gp120-capturing NS-pulsed dendritic cells could induce antigen-specific T-cell response. Intranasal administration of fluorescein isothiocyanate-labeled nanospheres (NS) in mice proved that the particles were taken up into pulmonary dendritic cells. Analysis of mice receiving intranasal immunizations with HIV-NS revealed that the mice efficiently induced the antibodies against HIV-1 in the genital tract and specific cytotoxic T-cells in the spleen. These results suggest that the use of HIV-1-NS may provide a novel and promising approach for the induction of humoral and cellular immune responses to HIV-1. PMID- 15778966 TI - Hepatitis B vaccination in a school age population: a feasibility study. AB - There remains no consensus on whether to adopt a universal hepatitis B vaccination strategy in the United Kingdom, where the endemicity of hepatitis B virus (HBV) is considered to be very low in the general population. To assess the feasibility and acceptance of a school-based adolescent vaccination approach, 11 13 years old pupils in local secondary schools in the London Borough of Camden and Islington were contacted and offered a three-dose hepatitis B vaccination course using a 0, 1, and 12 months schedule. The adult dose of hepatitis B vaccine (Engerix B GlaxoSmithKline) containing 20 mug recombinant hepatitis B surface antigen (HBsAg) in 1 ml suspension was administered. This dosage is normally intended for adults and children older than 15 years of age, but can be administered in 10-15 years old children when compliance may be low, since a higher proportion of those vaccinated develop protective antibody levels following administration of only two doses of vaccine. Overall, a total of 528 pupils were contacted, with 122 (23%) consenting to be vaccinated. Of these, 117 (96%) received the complete three-dose regimen according to the schedule (four did not receive vaccine: three were non-attendees and one was previously vaccinated; one withdrew following a flu-like adverse event). The results of this study show that it is feasible and practical to administer hepatitis B vaccination to adolescents in a school setting, and that it is possible to achieve high rates of uptake for the complete three-dose course among adolescents. However, in order to attain and sustain high coverage rates among pupils, this would require additional general health promotion, including health education and provision of information, targeting of teachers, pupils, and parents in order to increase participation in a school-based hepatitis B vaccination programme. A further requirement includes the availability of good local health support within schools so as to allow for an efficient vaccine delivery system to maximize vaccination in this setting. PMID- 15778967 TI - Automated computation of low-energy pathways for complex rearrangements in proteins: application to the conformational switch of Ras p21. AB - The computation of minimum energy paths (MEPs) is an approach for gaining insight into protein conformational transitions that are too slow to be observed with unconstrained molecular dynamics simulations. MEPs have the advantage of providing the energy barrier of the rate-limiting step(s), allowing discrimination among different paths. Finding low-energy MEPs for complex transitions, such as those involving rearrangements of the backbone fold or repacking of buried side chains, has hitherto been unfeasible in a reliable, automated manner, the MEP often displaying unphysical behavior, such as the crossing of bonds. Here, this problem is addressed by combining a counterintuitive procedure for generating an initial guess of the path, in which all side chains are shrunk, with the conjugate peak refinement (CPR) method. The effectiveness of the approach is tested on the conformational switch in Ras p21. This conformational transition involves some partial unfolding and re-folding, a process for which a multitude of pathways are likely to exist and for which a single MEP does not provide a complete description. However, this transition requires some sterically demanding rearrangements, thus testing the ability of a method to find low-energy pathways free of structurally unphysical events. This is achieved by the present approach, which finds a path whose rate-limiting barrier is compatible with experiment. This demonstrates that the method can be used to compute plausible pathways for complex rearrangements in proteins in an automated manner that is unbiased by external driving constraints. PMID- 15778968 TI - Differential expression of RARbeta isoforms in the mouse striatum during development: a gradient of RARbeta2 expression along the rostrocaudal axis. AB - The retinoic acid receptor RARbeta is highly expressed in the striatum of the ventral telencephalon. We studied the expression pattern of different RARbeta isoforms in the developing mouse striatum by in situ hybridization. We found a differential ontogeny of RARbeta2 and RARbeta1/3 in embryonic day (E) 13.5 lateral ganglionic eminence (striatal primordium). RARbeta2 mRNA was detected primarily in the rostral and ventromedial domains, whereas RARbeta1/3 mRNAs were enriched in the caudal and dorsolateral domains. Notably, by E16.5, a prominent decreasing gradient of RARbeta2 mRNA was present in the developing striatum along the rostrocaudal axis, i.e., RARbeta2 was expressed at higher levels in the rostral than the caudal striatum. No such gradient was found for RARbeta1/3 and RARbeta3 mRNAs. The rostrocaudal RARbeta2 gradient gradually disappeared postnatally and was absent in the adult striatum. The differential expression pattern of RARbeta isoforms in the developing striatum may provide an anatomical basis for differential gene regulation by RARbeta signaling. PMID- 15778969 TI - Profiles of HCV core protein and viremia in chronic hepatitis C: possible protective role of core antigen in liver damage. AB - The relation between HCV core antigen and HCV RNA has been confirmed in patients with chronic hepatitis C and a parallel behavior of the two markers has been described in early kinetics analysis during antiviral therapy. Variations of the core antigen to HCV RNA ratio have been reported in individual patients and the existence of nucleocapsid particles, not always associated with viral genomes, have been described. To assess the characteristics of HCV core antigen reactivity in relation to viremia in patients with different clinical profiles, 233 patients with chronic hepatitis C were studied serially. Group A included 54 asymptomatic HCV carriers, group B included 8 viremic patients with biochemical long-term response after antiviral therapy, while group C was composed of 171 patients with chronic liver disease and 75 were treated with combination therapy. Core antigen levels were not significantly different in the three groups of patients and a wide range of antigenic reactivity was observed in individual patients. A close relationship was observed between core antigen and HCV RNA, although their ratio was significantly higher in biochemical long-term responders (group B), compared to the other groups (P < 0.05). Physicochemical characterization of core antigen reactivity by equilibrium CsCl density gradient identified two distinct peaks migrating at 1.08-1.12 g/ml CsCl density and at 1.18-1.31 CsCl density, respectively. The first one, corresponding to the lipid-associated fraction, contained higher amounts of core antigen reactivity and was associated with clinical remission of liver damage, while the second peak, corresponding to naked nucleocapsids, was observed mainly in sera with active disease. In conclusion, a close relationship between core and HCV RNA was documented both in treated and untreated patients. The finding of an excess of lipid-associated core particles in a subset of viremic patients without biochemical activity of liver disease suggests their protective effect in liver cell damage. PMID- 15778971 TI - Linking tumor cell cytotoxicity to mechanism of drug action: an integrated analysis of gene expression, small-molecule screening and structural databases. AB - An integrated, bioinformatic analysis of three databases comprising tumor-cell based small molecule screening data, gene expression measurements, and PDB (Protein Data Bank) ligand-target structures has been developed for probing mechanism of drug action (MOA). Clustering analysis of GI50 profiles for the NCI's database of compounds screened across a panel of tumor cells (NCI60) was used to select a subset of unique cytotoxic responses for about 4000 small molecules. Drug-gene-PDB relationships for this test set were examined by correlative analysis of cytotoxic response and differential gene expression profiles within the NCI60 and structural comparisons with known ligand-target crystallographic complexes. A survey of molecular features within these compounds finds thirteen conserved Compound Classes, each class exhibiting chemical features important for interactions with a variety of biological targets. Protein targets for an additional twelve Compound Classes could be directly assigned using drug-protein interactions observed in the crystallographic database. Results from the analysis of constitutive gene expressions established a clear connection between chemo-resistance and overexpression of gene families associated with the extracellular matrix, cytoskeletal organization, and xenobiotic metabolism. Conversely, chemo-sensitivity implicated overexpression of gene families involved in homeostatic functions of nucleic acid repair, aryl hydrocarbon metabolism, heat shock response, proteasome degradation and apoptosis. Correlations between chemo-responsiveness and differential gene expressions identified chemotypes with nonselective (i.e., many) molecular targets from those likely to have selective (i.e., few) molecular targets. Applications of data mining strategies that jointly utilize tumor cell screening, genomic, and structural data are presented for hypotheses generation and identifying novel anticancer candidates. PMID- 15778970 TI - Monitoring low cytomegalovirus viremia in transplanted patients by a real-time PCR on plasma. AB - Until recently, human cytomegalovirus (hCMV) infection and anti-CMV treatment in transplanted patients have been monitored essentially by pp65 antigenemia, which is time-consuming and requires experienced operators. For the last two years, pp65 antigenemia levels have tended to be lower than previously in our laboratory, which could be due to better monitoring of CMV-related risk. Results obtained by real-time PCR with a LightCycler instrument or by pp65 antigen assay were compared on 145 serial samples from bone marrow or kidney transplant recipients under the usual conditions of our laboratory. CMV DNA was extracted from plasma and quantified by using primers and probes directed to HXFL4 gene. The plasma CMV DNA load was measured by using a standard curve constructed with a commercially available quantified CMV DNA suspension. Among the 145 samples, 139 showed a pp65 antigen which was negative or lower than 20 positively stained cells per 200,000 leukocytes. In the patients with positive pp65 antigenemia, the corresponding values of CMV DNA copy number/ml were significantly higher than those observed in patients without antigenemia (P < 0.001). CMV DNA was detected from 4 up to 52 days before pp65 antigen. Elsewhere, between two dates at which pp65 antigen was positive, intermediate PCR results could be positive while the pp65 antigen was negative. This real-time quantitative PCR assay is a rapid technique adapted to monitor plasma CMV DNA in transplant setting, even for low viremia. PMID- 15778972 TI - Variable expression of human myeloid specific nuclear antigen MNDA in monocyte lineage cells in atherosclerosis. AB - MNDA (human myeloid nuclear differentiation antigen) is expressed in specific lineages of hematopoietic cells and most notably at high levels in macrophages at sites of inflammation. MNDA and related proteins appear to modulate the activity of transcription factors and in some cases have a role in mediating cell death. The expression of MNDA was characterized in normal and diseased human aorta. MNDA positive cells double labeled for CD68 in all tissue examined. Twenty percent of normal aortas were negative or contained rare MNDA positive cells while other normal aorta contained more frequent positive cells. In atherosclerotic aorta, the number of MNDA positive cells increased with progression of disease. In normal and early lesions, MNDA positive cells adjacent to the endothelium generally displayed a strong MNDA reactivity associated with small amount of CD68 reactive cytoplasm. In the same sections, MNDA positive cells at increasing distances from the endothelium displayed lower MNDA reactivity and were associated with larger amounts of CD68 reactive cytoplasm. Foam cells in fatty streaks exhibited MNDA reactivity that ranged from strong to weak or negative. In advanced lesions, cells in the shoulder and those in fibrous tissue surrounding an atheroma were highly reactive for MNDA. However, only a fraction of the CD68 positive foam cells near the lipid core under the cap and shoulder contained MNDA reactivity. The variation in MNDA expression appeared to change with phenotypic specialization of monocytes in atherosclerosis consistent with its association with inflammation and suspected roles in regulating gene expression or in mediating cell death. PMID- 15778973 TI - Relationship between drug resistance mutations, plasma viremia, and CD4+ T-cell counts in patients with chronic HIV infection. AB - Transmission of drug-resistant viruses has been shown to be associated with lower virus replication capacity and higher CD4+ cell counts in recent human immunodeficiency virus (HIV) seroconvertors. The impact of drug resistance mutations on CD4 cell counts in chronically HIV-infected patients has not been examined. A total of 825 patients whose plasma specimens were submitted to a reference laboratory for genotypic testing from 1999 to 2002 were analyzed. There was no significant difference in the median CD4+ cell count when comparing 63 drug-naive and 762 treatment-experienced patients [399 (IQR, 141-525) vs. 319 (IQR, 174-521); P = 0.8]. In contrast, the median viral load was significantly higher in drug-naive than in pre-treated patients [4.6 (IQR, 4.1-5.25) vs. 4.1 (IQR, 3.4-4.7) logs; P < 0.0001]. Overall, drug resistance mutations appeared in 81% of patients, with a median number of 9 (IQR, 5-14). The rate of drug resistance genotypes was 9.5% for drug-naive patients and 86.7% for pre-treated individuals. In the univariate analysis, a lower viral load (P < 0.0001), the presence of drug-resistant viruses (P = 0.038), and specific mutations in the reverse transcriptase (RT) gene [presence of M184V (P = 0.016) or K70R (P < 0.0001), and lack of L74V (P < 0.003)] were all associated with higher CD4+ counts. However, in the multivariate analyses, only a lower viral load and the presence of K70R were significantly associated with higher CD4+ cell counts. In summary, drug-resistant viruses are associated with lower viral loads, but after adjusting for plasma viremia, subjects carrying drug-resistant viruses do not show significantly higher CD4 cell counts. Thus, keeping on treatment HIV infected individuals failing virologically and harboring drug-resistant viruses may ameliorate their immunological deterioration until new drugs became available. PMID- 15778974 TI - Genetic diversity of JC virus in the Saami and the Finns: implications for their population history. AB - The JC virus (JCV) genotyping method was used to gain insights into the population history of the Saami and the Finns, both speaking Finno-Ugric languages and living in close geographic proximity. Urine samples from Saami and Finns, collected in northern and southern Finland, respectively, were used to amplify a 610-bp JCV-DNA region containing abundant type-specific mutations. Based on restriction site polymorphisms in the amplified fragments, we classified JCV isolates into one of the three superclusters of JCV, type A, B, or C. All 15 Saami isolates analyzed and 41 of 43 Finnish isolates analyzed were classified as type A, the European type, and two samples from Finns were classified as type B, the African/Asian type. We then amplified and sequenced a 583-bp JCV-DNA region from the type A isolates of Saami and Finns. According to type-determining nucleotides within the region, we classified type A isolates into EU-a1, -a2, or b. Most type A isolates from Saami were classified as EU-a1, while type A isolates from Finns were distributed among EU-a1, EU-a2, and EU-b. This trend in the JCV-genotype distribution was statistically significant. On a phylogenetic tree based on complete sequences, most of the type A isolates from Saami were clustered in a single clade within EU-a1, while those from Finns were distributed throughout EU-a1, EU-a2, and EU-b. These findings are discussed in the context of the population history of the Saami and the Finns. This study provides new complete JCV DNA sequences derived from populations of anthropological interest. PMID- 15778975 TI - Gene trap screening as an effective approach for identification of Wnt-responsive genes in the mouse embryo. AB - In this study, we examined whether gene trap methodology, which would be available for systematic identification and functional analysis of genes, is effective for screening of Wnt-responsive genes during mouse development. We screened out two individual clones among 794 gene-trapped embryonic stem cell lines by their in vitro response to WNT-3A proteins. One gene was mainly expressed in the ductal epithelium of several developing organs, including the kidney and the salivary glands, and the other gene was expressed in neural crest cells and the telencephalic flexure. The spatial and temporal expression of these two genes coincided well with that of several Wnt genes. Furthermore, the expression of these two genes was significantly decreased in embryos deficient for Wnts or in cultures of embryonic tissues treated with a Wnt signal inhibitor. These results indicate that the gene trap is an effective method for systematic identification of Wnt-responsive genes during embryogenesis. PMID- 15778976 TI - Factors accelerating liver fibrosis progression in renal transplant patients receiving ribavirin monotherapy for chronic hepatitis C. AB - In untreated hepatitis virus (HCV)-positive renal transplant patients, the rate of liver fibrosis progression is low. In contrast, in those treated by ribavirin monotherapy, liver fibrosis score increased significantly after only 1 year of ribavirin monotherapy. The aim of this study was to identify the factors that might contribute to accelerate liver fibrosis progression in this population. Eleven patients were included in the study. Intrahepatic transforming growth factor (TGF)-beta, interferon (IFN)-gamma, and interleukin (IL)-10 mRNA quantification determined by real-time reverse transcription-polymerase chain reaction (RT-PCR) were similar before and after ribavirin therapy. The number of amino acid substitutions observed in the hypervariable region (HVR)-1 of the HCV genome between baseline and 1 year after ribavirin monotherapy was low, i.e., 3 (1-11) amino acid substitutions, suggesting the absence of a high selection pressure induced by ribavirin. In contrast, due to ribavirin-induced hemolysis, there was a significant increase in serum ferritin levels (P = 0.02) and in intrahepatic iron deposition (P = 0.04). Transferrin level and total iron-binding capacity decreased significantly during ribavirin monotherapy (P = 0.004). The increased liver fibrosis observed in renal transplant patients receiving ribavirin monotherapy could be related to ribavirin-induced anemia. Severe chronic hemolysis is responsible for iron overload, liver iron deposition, and an acceleration in the progression of liver fibrosis. PMID- 15778977 TI - Epstein-Barr virus (EBV)-encoded BARF1 gene is expressed in nasopharyngeal carcinoma and EBV-associated gastric carcinoma tissues in the absence of lytic gene expression. AB - The BARF1 gene is located in the BamHI-A fragment of the Epstein-Barr virus (EBV) genome, encodes 221 amino acids, and has activity as an oncogene. Several reports have demonstrated that BARF1 is expressed in the tissues of various EBV associated epithelioid malignancies. However,BARF1 is thought to be a lytic gene, since its expression is induced upon induction of the lytic cycle in Burkitt's lymphoma cell lines. Therefore, the possibility cannot be excluded that BARF1 expression in EBV-associated epithelioid malignancies reflects spontaneous induction of the lytic cycle in carcinoma cells. The present study aimed to clarify whether BARF1 was expressed as a latent gene or a lytic gene in epithelioid malignancies. Quantitative real-time RT-PCR assay revealed that BARF1 was highly expressed in nasopharyngeal carcinoma (NPC) and EBV-positive gastric carcinoma tissues in the absence of expression of lytic genes. On the other hand, BARF1 protein was detectable only in two of seven NPC tissue samples by immunoblot analysis. Analysis of BARF1-transfected CNE1 cells revealed that BARF1 was quickly secreted into culture medium and was hardly detectable in the cell lysate, which would account for why some NPC tissues were negative for BARF1 protein expression even though they were strongly positive forBARF1 expression at the transcriptional level. The present findings indicate that BARF1 is expressed in NPC and EBV-positive gastric carcinoma tissues as a latent gene and suggest that BARF1 plays a role in the pathogenesis of these malignancies. PMID- 15778978 TI - Hierarchical nature of morphological integration and modularity in the human posterior face. AB - Morphological integration and modularity are important points of intersection between evolution and the development of organismal form. Identification and quantification of integration are also of increasing paleoanthropological interest. In this study, the "posterior face," i.e., the mandibular ramus and its integration with the associated midline and lateral basicranium, is analyzed in lateral radiographs of 144 adult humans from three different geographic regions. The null hypothesis of homogenously pervasive morphological integration among "posterior-face" components is tested with Procrustes geometric morphometrics, partial least squares, and singular warps analysis. The results reveal statistically significant differences in integration. Only loose integrative relationships are found between midline and lateral components of the basicranium, which may indicate the presence of at least two different basicranial modules. This modularity can be interpreted in terms of spatiotemporal dissociation in the development of those basicranial structures, and gives support to hypotheses of independent phylogenetic modifications at the lateral and midline basicranium in humans. In addition, morphological integration was statistically significantly stronger between the middle cranial fossa and the mandibular ramus than between the ramus and the midline cranial base. This finding confirms previous hypotheses of a "petroso-mandibular unit," which could be a developmental consequence of well-known phylogenetic modifications in coronal topology of the posterior face and base in hominoid evolution, related to middle cranial fossa expansion. This unit could be involved in later evolutionary tendencies in the hominid craniofacial system. PMID- 15778979 TI - Endogenously expressed estrogen receptors mediate neuroprotection in hippocampal cells (HT22). AB - Discovery of estrogen receptors (ER) in the central nervous system and the ability of estrogens to modulate neural circuitry and act as neurotrophic factors, suggest a therapeutic role of this steroid. To gain better understanding of the specificity and cellular mechanisms involved in estrogen-mediated neuroprotection, a mouse hippocampal neuronal cell line (HT22) was evaluated. Earlier reports indicated this cell line was devoid of ERs. Contrary to these findings, characterization of HT22 cells using RT-PCR, immunoblot, immunocytochemical, and radioligand binding techniques revealed endogenous expression of ER. The predominant subtype appeared to be ERalpha with functional activity confirmed using an ERE-tk-luciferase assay. The ability of an ER antagonist, ICI-182780, to block the neuroprotective effects of estrogens confirmed ER was involved mechanistically in neuroprotection. In conclusion, HT22 cells express functional ERalpha or a closely related ER enabling this cell line to be used to profile estrogens for neuroprotective properties acting via an ER dependent mechanism. PMID- 15778980 TI - Human papillomavirus genotypes and their association with cervical neoplasia in a cohort of Western Australian women. AB - Human papillomavirus (HPV) is known to be the cause of almost all cervical cancers. The genotypes have been classified into high and low risk types according to their oncogenic potential. However, data for many of the genotypes are limited and some (HPV-26, 53, and 66) have no agreed status. A study was undertaken to determine the HPV genotype distribution in women of Western Australia and the association with cervical neoplasia. Liquid based cervical samples from a cohort of 282 Western Australian women were tested for HPV DNA by PCR followed by DNA sequencing to determine HPV genotypes. HPV-53 and HPV-16 were the most common genotypes found in this population. In addition 86 archived liquid based cervical samples from women with cervical intraepithelial neoplasia grades 1-3 (CIN 1-3) were tested for HPV DNA. Also 32 archived paraffin biopsy samples from women with squamous cell carcinoma were also tested. HPV-16 was the most common genotype found in these samples. Of the cohort of Western Australian women tested, 27% were found to contain HPV and approximately half of these contained known high-risk HPV genotypes, but only 30% of these were types 16 or 18. The data from this study indicate that HPV-53 is not oncogenic based on an R value and odds ratio (OR) of zero. The data also suggest that HPV-73 may be oncogenic, while HPV-66 is unlikely to be. Two high-risk HPV genotypes that are associated with the Asian region (HPV-52 and HPV-58) were found in Western Australian women suggesting a possible epidemiological link between women in these countries. PMID- 15778981 TI - Importance of cooperation and affiliation in the evolution of primate sociality. AB - The idea that competition and aggression are central to an understanding of the origins of group-living and sociality among human and nonhuman primates is the dominant theory in primatology today. Using this paradigm, researchers have focused their attention on competitive and aggressive behaviors, and have tended to overlook the importance of cooperative and affiliative behaviors. However, cooperative and affiliative behaviors are considerably more common than agonistic behaviors in all primate species. The current paradigm often fails to explain the context, function, and social tactics underlying affiliative and agonistic behavior. Here, we present data on a basic question of primate sociality: how much time do diurnal, group-living primates spend in social behavior, and how much of this time is affiliative and agonistic? These data are derived from a survey of 81 studies, including 28 genera and 60 species. We find that group living prosimians, New World monkeys, Old World monkeys, and apes usually devote less than 10% of their activity budget to active social interactions. Further, rates of agonistic behaviors are extremely low, normally less than 1% of the activity budget. If the cost to the actors of affiliative behavior is low even if the rewards are low or extremely variable, we should expect affiliation and cooperation to be frequent. This is especially true under conditions in which individuals benefit from the collective environment of living in stable social groups. PMID- 15778982 TI - Regulation by the extracellular matrix (ECM) of prolactin-induced alpha s1-casein gene expression in rabbit primary mammary cells: role of STAT5, C/EBP, and chromatin structure. AB - The aim of the present study was to understand how the extracellular matrix (ECM) regulates at the gene level the prolactin (Prl)-induced signal transducer and activator of transcription 5 (STAT5)-dependent expression of the alpha s1-casein gene in mammary epithelial cells. CCAAT enhancer binding proteins (C/EBPs) are assumed regulators of beta-casein gene expression. Rabbit primary mammary cells express alpha s1-casein gene when cultured on collagen and not on plastic. Similar C/EBPbeta, C/EBPdelta, STAT5, and Prl-activated STAT5 were found under all culture conditions. Thus the ECM does not act through C/EBPs or STAT5. This was confirmed by transfections of rabbit primary mammary cells by a construct sensitive to ovine prolactin (oPrl) and ECM (6i TK luc) encompassing STAT5 and C/EBP binding sites. The mutation of C/EBPs binding sites showed that these sites were not mandatory for Prl-induced expression of the construct. Interestingly, chromatin immunoprecipitation by the anti-acetylhistone H4 antibody (ChIP) showed that the ECM (and not Prl) maintained a high amount of histone H4 acetylation upstream of the alpha s1-casein gene especially at the level of a distal Prl- and ECM-sensitive enhancer. Alpha6 integrin (a membrane receptor of laminin, the principal active component of the mammary ECM) was found at the surface of cells cultured on collagen but not on plastic. In cells cultured on collagen in the presence of anti-alpha6 integrin antibody, Prl-induced transcription of the endogenous alpha s1-casein gene was significantly reduced, without modifying C/EBPs and STAT5. Besides, histone H4 acetylation was reduced. Thus, we propose that the ECM regulates rabbit alpha s1-casein protein expression by local modification of chromatin structure, independently of STAT5 and C/EBPs. PMID- 15778984 TI - Transgenic frogs expressing the highly fluorescent protein venus under the control of a strong mammalian promoter suitable for monitoring living cells. AB - To easily monitor living cells and organisms, we have created a transgenic Xenopus line expressing Venus, a brighter variant of yellow fluorescent protein, under the control of the CMV enhancer/chicken beta-actin (CAG) promoter. The established line exhibited high fluorescent intensity not only in most tissues of tadpoles to adult frogs but also in germ cells of both sexes, which enabled three dimensional imaging of fluorescing organs from images of the serial slices of the transgenic animals. Furthermore, by using this transgenic line, we generated chimeric animals by brain implantation and importantly, we found that the brain grafts survived and expressed Venus in recipients after development, highlighting the boundary between fluorescent and nonfluorescent areas in live animals. Thus, Venus-expressing transgenic frogs, tadpoles, and embryos would facilitate their use in many applications, including the tracing of the fluorescent cells after tissue/organ transplantation. PMID- 15778983 TI - Expression and antigenicity of virus-like particles of norovirus and their application for detection of noroviruses in stool samples. AB - Human noroviruses (NoVs), members of the genus Norovirus in the family Caliciviridae, are the leading agents of nonbacterial acute gastroenteritis worldwide. Human NoVs are currently divided into at least two genogroups, genogroup I (GI) and genogroup II (GII), each of which contains at least 14 and 17 genotypes. To explore the genetic and antigenic relationship among NoVs, we expressed the capsid protein of four genetically distinct NoVs, the GI/3 Kashiwa645 virus, the GII/3 Sanbu809 virus, the GII/5 Ichikawa754 virus, and the GII/7 Osaka10-25 virus in baculovirus expression system. An antigen enzyme-linked immunosorbent assay (ELISA) with hyperimmune serum against the four recombinant capsid proteins and characterized previously three capsid proteins derived from GI/1, GI/4, and GII/12 was developed to detect the NoVs antigen in stools. The antigen ELISA was highly specific to the homotypic strains, allowing assignment of a strain to a Norovirus genetic cluster within a genogroup. PMID- 15778985 TI - Inhibitors of ubiquitin-dependent proteolysis can delay programmed cell death of adult intersegmental muscles in the moth Manduca sexta. AB - In the moth Manduca sexta, intersegmental muscles (ISMs) undergo rapid programmed cell death (PCD) within 48 hr of adult emergence. ISM PCD involves ubiquitin dependent proteasomal degradation accompanied by the down-regulation of expression of actin genes and the up-regulation of degradative gene expression such as ubiquitin. Hemin chloride and N-acetyl-leu-leu-norleucinal (ALLN), both inhibitors of proteasomal activity, administered before adult emergence delayed PCD for up to 5 days in ISMs maintained from the larval stage, such as the dorsal internal medial muscle in abdominal segment 4 (DIM-A4). ISMs that developed during metamorphosis from respecified larval muscles such as the DIM-A2 were less dramatically affected. The increase in polyubiquitinated proteins and the decrease in actin mRNA expression accompanying maintained ISM PCD were delayed after inhibitor application. No changes were detected in respecified ISMs. These results reveal a regulatory role for proteasomal activity in an early stage of maintained ISM cell death. PMID- 15778986 TI - Development of a perfusion fed bioreactor for embryonic stem cell-derived cardiomyocyte generation: oxygen-mediated enhancement of cardiomyocyte output. AB - Cell transplantation is emerging as a promising new approach to replace scarred, nonfunctional myocardium in a diseased heart. At present, however, generating the numbers of donor cardiomyocytes required to develop and test animal models is a major limitation. Embryonic stem (ES) cells may be a promising source for therapeutic applications, potentially providing sufficient numbers of functionally relevant cells for transplantation into a variety of organs. We developed a single-step bioprocess for ES cell-derived cardiomyocyte production that enables both medium perfusion and direct monitoring and control of dissolved oxygen. Implementation of the bioprocess required combining methods to prevent ES cell aggregation (hydrogel encapsulation) and to purify for cardiomyocytes from the heterogeneous cell populations (genetic selection), with medium perfusion in a controlled bioreactor environment. We used this bioprocess to investigate the effects of oxygen on cardiomyocyte generation. Parallel vessels (250 mL culture volume) were run under normoxic (20% oxygen tension) or hypoxic (4% oxygen tension) conditions. After 14 days of differentiation (including 5 days of selection), the cardiomyocyte yield per input ES cell achieved in hypoxic vessels was 3.77 +/- 0.13, higher than has previously been reported. We have developed a bioprocess that improves the efficiency of ES cell-derived cardiomyocyte production, and allows the investigation of bioprocess parameters on ES cell derived cardiomyogenesis. Using this system we have demonstrated that medium oxygen tension is a culture parameter that can be manipulated to improve cardiomyocyte yield. PMID- 15778987 TI - Correlation for the partition behavior of proteins in aqueous two-phase systems: effect of surface hydrophobicity and charge. AB - Correlations to describe the effect of surface hydrophobicity and charge of proteins with their partition coefficient in aqueous two-phase systems were investigated. Polyethylene glycol (PEG) 4000/phosphate, sulfate, citrate, and dextran systems in the presence of low (0.6% w/w) and high (8.8% w/w) levels of NaCl were selected for a systematic study of 12 proteins. The surface hydrophobicity of the proteins was measured by ammonium sulfate precipitation as the inverse of their solubility. The hydrophobicity values measured correlated well with the partition coefficients, K, obtained in the PEG/salt systems at high concentration of NaCl (r = 0.92-0.93). In PEG/citrate systems the partition coefficient correlated well with protein hydrophobicity at low and high concentrations of NaCl (r = 0.81 and 0.93, respectively). The PEG/citrate system also had a higher hydrophobic resolution than other systems to exploit differences in the protein's hydrophobicity. The surface charge and charge density of the proteins was determined over a range of pH (3-9) by electrophoretic titration curves; PEG/salt systems did not discriminate well between proteins of different charge or charge density. In the absence of NaCl, K decreased slightly with increased positive charge. At high NaCl concentration, K increased as a function of positive charge. This suggested that the PEG-rich top phase became more negative as the concentration of NaCl in the systems increased and, therefore, attracted the positively charged proteins. The effect of charge was more important in PEG/dextran systems at low concentrations of NaCl. In the PEG/dextran systems at lower concentration of NaCl, molecular weight appeared to be the prime determinant of partition, whereas no clear effect of molecular weight could be found in PEG/salt systems. PMID- 15778988 TI - Interactions of lysozyme in guanidinium chloride solutions from static and dynamic light-scattering measurements. AB - The interactions of partially unfolded proteins provide insight into protein folding and protein aggregation. In this work, we studied partially unfolded hen egg lysozyme interactions in solutions containing up to 7 M guanidinium chloride (GdnHCl). The osmotic second virial coefficient (B(22)) of lysozyme was measured using static light scattering in GdnHCl aqueous solutions at 20 degrees C and pH 4.5. B(22) is positive in all solutions, indicating repulsive protein-protein interactions. At low GdnHCl concentrations, B(22) decreases with rising ionic strength: in the absence of GdnHCl, B(22) is 1.1 x 10(-3) mLmol/g(2), decreasing to 3.0 x 10(-5) mLmol/g(2) in the presence of 1 M GdnHCl. Lysozyme unfolds in solutions at GdnHCl concentrations higher than 3 M. Under such conditions, B(22) increases with ionic strength, reaching 8.0 x 10(-4) mLmol/g(2) at 6.5 M GdnHCl. Protein-protein hydrodynamic interactions were evaluated from concentration dependent diffusivity measurements, obtained from dynamic light scattering. At moderate GdnHCl concentrations, lysozyme interparticle interactions are least repulsive and hydrodynamic interactions are least attractive. The lysozyme hydrodynamic radius was calculated from infinite-dilution diffusivity and did not change significantly during protein unfolding. Our results contribute toward better understanding of protein interactions of partially unfolded states in the presence of a denaturant; they may be helpful for the design of protein refolding processes that avoid protein aggregation. PMID- 15778991 TI - Role of the intracellular domains of LRP5 and LRP6 in activating the Wnt canonical pathway. AB - LDL-receptor related proteins 5 and 6 (LRP5/6) are co-receptors of Frizzled receptors that mediate Wnt-induced activation of the transcription factor family TCF/LEF-1. Even though LRP5 and LRP6 are highly homologous, LRP6, but not LRP5, is expressed primarily in the nervous system and deletion of the LRP6 gene results in significant brain abnormalities, while deletion of LRP5 results in primarily decreased bone density. Additionally, the exact function of LRP5 and LRP6 have not been clearly defined, although it is clear that they both play key roles in the Wnt canonical pathway. In this study the role of the intracellular domains of LRP5/6 in mediating Wnt signaling was examined. In the absence of exogenous Wnt 3a, full-length (FL) LRP6, but not LRP5, increased TCF/LEF-1 transcriptional activity, however both significantly potentiated Wnt 3a-induced TCF/LEF-1 activation. In contrast to the findings with the FL constructs, the intracellular domains (membrane-anchored and cytosolic) of both LRP5 and LRP6 significantly increased TCF/LEF-1 activation in the absence of Wnt 3a, and potentiated the Wnt 3a-induced decrease in beta-catenin phosphorylation, increase in free beta-catenin levels and the increase in TCF/LEF-1 activity. These findings demonstrate that: (1) LRP5 and LRP6 differentially modulate TCF/LEF-1 activation in the absence of Wnt 3a and (2) the intracellular C-terminal domains of LRP5/6 potentiate Wnt 3a-induced TCF/LEF-1 activation whether or not they are membrane-anchored. These findings provide significant new insights into the roles of LRP5/6 in modulating canonical Wnt signaling. PMID- 15778992 TI - Studies on epidermal growth factor receptor signaling in vertebrate limb patterning. AB - The epidermal growth factor receptor (EGFR) regulates multiple patterning events in Drosophila limb development, but its role in vertebrate limb morphogenesis has received little attention. The EGFR and several of its ligands are expressed in developing vertebrate limbs in manners consistent with potential patterning roles. To gain insight into functions of EGFR signaling in vertebrate limb development, we expressed a constitutively active EGFR in developing chick limbs in ovo. Expression of activated EGFR causes pre- and postaxial polydactyly, including mirror-image-type digit duplication, likely due to induction of ectopic expression and/or modulation of genes involved in anterior-posterior (AP) patterning such as Sonic hedgehog (Shh), dHand, Patched (Ptc), Gli3, Hoxd13, Hoxd11, bone morphogenetic protein 2 (Bmp2), Gremlin, and FGF4. Activation of EGFR signaling dorsalizes the limb and alters expression of the dorsal-ventral (DV) patterning genes Wnt7a, Lmx, and En1. Ectopic and/or extended FGF8 expressing apical ectodermal ridges (AERs) are also seen. Interdigital regression is inhibited and the digits fail to separate, leading to syndactyly, likely due to antiapoptotic and pro-proliferative effects of activated EGFR signaling on limb mesoderm, and/or attenuation of interdigital Bmp4 expression. These findings suggest potential roles for EGFR signaling in AP and DV patterning, AER formation, and cell survival during limb morphogenesis. PMID- 15778993 TI - Signaling through FGF receptor-2 is required for lens cell survival and for withdrawal from the cell cycle during lens fiber cell differentiation. AB - Fibroblast growth factors (FGFs) play important roles in many aspects of development, including lens development. The lens is derived from the surface ectoderm and consists of an anterior layer of epithelial cells and elongated, terminally differentiated fiber cells that form the bulk of the tissue. FGF signaling has been implicated in lens induction, proliferation, and differentiation. To address the role of FGFs in lens development, we inactivated FGF receptor-2 (Fgfr2) using a Cre transgene that is expressed in all prospective lens cells from embryonic day 9.0. Inactivation of Fgfr2 shows that signaling through this receptor is not required for lens induction or for the proliferation of lens epithelial cells. However, Fgfr2 signaling is needed to drive lens fiber cells out of the cell cycle during their terminal differentiation. It also contributes to the normal elongation of primary lens fiber cells and to the survival of lens epithelial cells. PMID- 15778994 TI - Development and validation of algorithms for measuring G-protein coupled receptor activation in cells using the LSC-based imaging cytometer platform. AB - BACKGROUND: A cell-based assay system (Transfluor) has been developed for measurement of G-protein coupled receptor (GPCR) activity by using cells transfected to express a fusion protein of arrestin plus green fluorescent protein (GFP) and the target GPCR. Upon agonist stimulation, the arrestin-GFP translocates to and binds the activated GPCR at the plasma membrane. The receptor/arrestin-GFP complexes then localize in clathrin-coated pits and/or intracellular vesicles. This redistribution of arrestin-GFP into condensed fluorescent spots is useful for visually monitoring the active status of GPCRs and its quantitation is possible with certain types of digital image analysis systems. METHODS: We designed two lines of image processing algorithms to carry out quantitative measurement of the arrestin-GFP movement on an inverted version of laser scanning cytometry (iCyte) as an imaging platform. We used a cell line expressing arrestin-GFP and the wild-type beta2-adrenergic receptor or a modified version of this receptor with enhanced affinity for arrestin. Each cell line was challenged with various concentrations of agonist. RESULTS: A dose-dependent signal was measured and half-maximal effective concentration values were obtained that agreed well with results determined by other methods previously reported. CONCLUSIONS: The results indicate that the combination of Transfluor, iCyte, and our algorithms is suitable for robust and pharmacologically relevant GPCR ligand exploration. PMID- 15778995 TI - Cloning and developmental expression of Xenopus Enabled (Xena). AB - Regulation of actin dynamics, organization, and interaction with cell surface adhesion proteins is critical for tissue morphogenesis during development. The Ena/VASP family of actin-binding proteins function in several cellular processes that involve dynamic regulation of the actin cytoskeleton, including axon guidance, platelet aggregation, cell migration, and cell adhesion. The vertebrate Ena/VASP family is composed of three genes: Ena (Enabled), VASP (Vasodilator Stimulated Phosphoprotein), and Evl (Ena/VASP-Like). To better understand the role of Ena/VASP proteins during vertebrate development, we have cloned and characterized the developmental expression of Ena in Xenopus laevis. Analysis of the temporal expression of Xenopus Ena (Xena) demonstrates that multiple isoforms of Xena are detected throughout embryogenesis and that the presence of different isoforms is developmentally regulated. In situ hybridization analyses reveal that Xena is broadly expressed throughout development. During gastrulation and neurulation, Xena is detected in the neuroepithelium, notochord, and somites. In tadpoles, Xena expression is restricted to dorsal regions of the brain, whereas it is expressed at lower levels throughout the spinal cord. Xena expression is also detected in the notochord, myotome, heart, pronephros, and cranial placodes, including the olfactory and otic placodes. Analysis of the subcellular localization of Xena using a GFP fusion protein revealed that Xena localizes to adherens junctions and focal adhesions in Xenopus animal caps and NIH3T3 fibroblasts, respectively. These results define spatiotemporal windows in which Xena may function during early Xenopus development to modulate actin-dependent processes such as cell adhesion and migration. PMID- 15778996 TI - Planar cell polarity: heading in the right direction. AB - Epithelial cells are patterned not only along their apical-basolateral axis, but also along the plane of the epithelial sheet; the latter event is regulated by the planar cell polarity (PCP) pathway. PCP regulates diverse outputs, such as the distal placement of a hair in all cells of the Drosophila wing, and convergent extension movements during gastrulation in the vertebrate embryo. This primer describes the molecular mechanisms that initiate and establish PCP, as well as biochemical pathways that translate PCP signaling to cell type-specific patterning events. The primer concludes with a discussion of current topics in the field with two PCP researchers, Matt Kelley, Ph.D., and Helen McNeill, Ph.D. PMID- 15778998 TI - Overexpression of GSTA2 protects against cell cycle arrest and apoptosis induced by the DNA inter-strand crosslinking nitrogen mustard, mechlorethamine. AB - The effectiveness of bifunctional alkylating nitrogen mustard compounds in chemotherapy is related to their ability to form DNA inter-strand crosslinks. Patients exposed to DNA inter-strand crosslinking (ICL) agents subsequently experience an elevated incidence of myelodysplastic syndromes (MDS) and MDS related acute myeloid leukemia. Fanconi's anemia (FA) patients are deficient in the repair of crosslink DNA damage and they experience a high incidence of MDS. These observations indicate that hematopoietic cells are specific target for the transforming effects of DNA crosslinking damage. Changes in transcript levels were characterized in human hematopoietic cells occurring in response to the nitrogen mustard, mechlorethamine (HN2), but not in response to monofunctional analogs. Only modest changes in a few gene transcripts were detected in HL60 cells exposed to levels of HN2 tittered to maximal dose that caused growth suppression with minimal cell death and allowed eventual resumption of normal cell growth. Under conditions of transient growth suppression, a subset of glutathione-S-transferase (GST) isoenzyme genes was consistently upregulated three to fourfold by HN2, but not by monofunctional analogs. Subsequent efforts to confirm the changes detected by microarray analyses revealed an unexpected dependence on treatment conditions. The GST alpha class A2 subfamily member transcripts were upregulated 24 h after a 1 h exposure to HN2 that caused an extensive, but transient block in late S/G2 cell cycle phase, but were minimally altered with continuous exposure. The 1-h exposure to HN2 caused a transient late S/G2 cell cycle arrest in both the HL-60 cell line and the Colo 320HSR human colon cancer cell line. Overexpression of GSTA2 by transient transfection protected Colo 320HSR cells against both cycle arrest and apoptosis following exposure to HN2. Overexpression of GSTA2 in Colo 320HSR cells induced after exposure to HN2 did not alter cycle arrest or apoptosis. The results indicate that human GSTA2 facilitates the protection of cells from HN2 damage and not repair. Our results are consistent with the possibility that GSTA2 polymorphisms, variable isoenzyme expression, and variable induced expression may be factors in the pathogenesis of MDS. PMID- 15778999 TI - SP100B is a repressor of gene expression. AB - Mammalian cell nuclei exhibit discrete sites where specific proteins characteristically localize. PML nuclear bodies (PML NBs) (nuclear domain 10s (ND10s)) are the primary localization site for the promyelocytic leukemia (PML) protein and the SP100 autoantigen. The observations that some PML and SP100 isoforms can function as transcriptional regulators, that both the size and number of PML bodies increase in response to interferon treatment, and that many mammalian viruses encode proteins that mediate disruption of PML bodies suggest that these sites suppress viral infection, perhaps by repressing viral gene expression. We hypothesized that a component of PML NBs functions as a repressor of gene expression. To test this hypothesis, we characterized the effect of PML or SP100 isoforms on expression of transfected reporter genes. PML-I, PML-VI, and SP100A did not repress reporter gene expression. In contrast, SP100B repressed reporter gene expression, especially under conditions in which the reporter gene expression was elevated by a viral transactivator or addition of trichostatin A to the culture medium. The SP100B DNA binding domain was required for repression. SP100B had no detectable effect on the amount, methylation pattern, or topological form of plasmid DNA in the nuclei of transfected cells. The demonstrated repressive activity of SP100B supports the hypothesis that SP100B is a component of an innate immune response that represses expression of ectopic DNA. PMID- 15779000 TI - Switching stimulation patterns improves performance of paralyzed human quadriceps muscle. AB - The clinical efficacy of functional electrical stimulation (FES) is limited by the rapid onset of fatigue. Functional electrical stimulation applications typically stimulate skeletal muscles with constant-frequency trains (CFTs). Our laboratory has identified trains that we call doublet-frequency trains (DFTs) and that produce greater forces than CFTs, but more fatigue during repetitive activation than comparable CFTs. The purpose of this study was to see whether a series of CFTs followed by DFTs would reach a targeted isometric peak force more times than either train type alone during repetitive isometric activation of the paralyzed quadriceps muscles of subjects with spinal cord injuries (SCI). The combination of CFTs followed by DFTs reached the targeted isometric force 14% more often than the CFTs alone and 18% more often than the DFTs alone. These findings confirm that switching train types may be a useful strategy to offset the rapid fatigue of the functionally important quadriceps muscle that persons with SCI experience when using FES. PMID- 15779002 TI - Direct inhibition of interleukin-2 receptor alpha-mediated signaling pathway induces G1 arrest and apoptosis in human head-and-neck cancer cells. AB - Overexpression of the interleukin-2 receptor (IL-2R) alpha chain in tumor cells is associated with tumor progression and a poor patient prognosis. IL-2Ralpha is responsible for the high affinity binding of the receptor to IL-2, leading to activation of several proliferative and anti-apoptotic intracellular signaling pathways. We have previously shown that human squamous cell carcinoma of a head and-neck line (PCI-13) genetically engineered to overexpress IL-2Ralpha exhibit increased transforming activity, proliferation, and drug resistance, compared to the vector control cells (J Cell Biochem 2003;89:824-836). In this study, we report that IL-2Ralpha(+) cells express high levels of total and phosphorylated Jak3 protein and are more resistant to apoptosis induced by a Jak3 inhibitor than the control LacZ cells. Furthermore, we used daclizumab, a monoclonal antibody specific to IL-2Ralpha, and determined the effects of IL-2Ralpha inhibition on cell cycle and apoptosis as well as the involvement of potential cell cycle and apoptosis regulatory proteins. We found that daclizumab induces G(1) arrest, associated with down-regulation of cyclin A protein, preferentially in IL 2Ralpha(+) cells, but not in LacZ cells. In addition, daclizumab activates apoptotic death program via Bcl-2 down-regulation preferentially in IL-2Ralpha(+) cells. Finally, daclizumab also sensitizes IL-2Ralpha(+) cells to other apoptotic stimuli, although the effect is moderate. These results indicate that daclizumab inhibits the proliferative potential of IL-2Ralpha(+) cells via inhibition of cell cycle progression and induction of apoptosis. PMID- 15779001 TI - Adhesion-dependent redistribution of MAP kinase and MEK promotes muscarinic receptor-mediated signaling to the nucleus. AB - The mitogen-activated protein kinases (MAPKs) are activated by extracellular signals, and translocate to the nucleus where they modulate transcription. Integrin-mediated cell adhesion to extracellular matrix (ECM) proteins is required for efficient transmission of MAPK-based signals initiated by growth factors. However, the modulation of G protein-coupled receptor (GPCR) signaling by adhesion is less well understood. In the present study, we assessed the impact of cell adhesion on MAPK activation by muscarinic M3 receptors. The muscarinic agonist carbachol more efficiently promoted stress fiber formation and tyrosine phosphorylation of focal adhesion-associated proteins in M3 receptor-expressing cells adherent to fibronectin or collagen type I, as compared to polylysine. Overall MAPK activation was robust in cells adherent to all three substrata. However, total levels of MAPK and mitogen-activated protein kinase kinase (MEK) in the nucleus were significantly greater in cells adherent to ECM proteins for 2.5 h, and levels of activated MAPK and MEK in the nuclei of these cells were higher following carbachol stimulation, relative to levels in cells adherent to polylysine. MEK inhibitors did not prevent adhesion-dependent translocation of MAPK and MEK to the nucleus, and increased nuclear phospho-MEK levels in carbachol-stimulated cells. The results suggest that adhesion of cells to ECM triggers the redistribution of MAPK and MEK to the nucleus, possibly as a result of the cytoskeletal rearrangements that accompany cell spreading. This may represent a mechanism for priming the nucleus with MEK and MAPK, leading to more rapid and pronounced increases in intranuclear phospho-MAPK upon GPCR stimulation. PMID- 15779003 TI - Voluntary muscle activation, contractile properties, and fatigability in children with and without cerebral palsy. AB - Cerebral palsy (CP) may lead to profound weakness in affected portions of the extremities and trunk. Knowing the mechanisms underlying muscle weakness will help to better design interventions for increasing force production in children with CP. This study quantified voluntary muscle activation, contractile properties, and fatigability of the quadriceps femoris and triceps surae in children with and without CP. Twelve children with CP (7-13 years) and 10 unaffected children (controls, 8-12 years) were assessed for (1) voluntary muscle activation during maximum voluntary isometric contractions (MVICs); (2) antagonist coactivation during agonist MVICs; (3) contractile properties, and (4) fatigability using electrically elicited tests. Children with CP were significantly weaker, had lower agonist voluntary muscle activation, and greater antagonist coactivation. In children with CP, the quadriceps normalized force frequency relationship (FFR) was shifted upward at low frequencies and was less fatigable than controls. No differences were seen between groups in the normalized FFR and fatigability of the triceps surae. In addition, no differences were seen in the sum of the time to peak tension and half-relaxation times between groups for either muscle. Because children with CP demonstrated large deficits in voluntary muscle activation, using voluntary contractions for strength training may not produce forces sufficient to induce muscle hypertrophy. Techniques such as enhanced feedback and neuromuscular electrical stimulation may be helpful for strengthening muscles that cannot be sufficiently recruited with voluntary effort. PMID- 15779004 TI - Imprinting centers, chromatin structure, and disease. AB - Two regions that best exemplify the role of genetic imprinting in human disease are the Prader-Willi syndrome/Angelman syndrome (PWS/AS) region in 15q11-q13 and the Beckwith-Wiedemann syndrome (BWS) region in 11p15.5. In both regions, cis acting sequences known as imprinting centers (ICs) regulate parent-specific gene expression bidirectionally over long distances. ICs for both regions are subject to parent-specific epigenetic marking by covalent modification of DNA and histones. In this review, we summarize our current understanding of IC function and IC modification in these two regions. PMID- 15779005 TI - MLL: how complex does it get? AB - The mixed lineage leukemia (MLL) gene encodes a very large nuclear protein homologous to Drosophila trithorax (trx). MLL is required for the proper maintenance of HOX gene expression during development and hematopoiesis. The exact regulatory mechanism of HOX gene expression by MLL is poorly understood, but it is believed that MLL functions at the level of chromatin organization. MLL was identified as a common target of chromosomal translocations associated with human acute leukemias. About 50 different MLL fusion partners have been isolated to date, and while similarities exist between groups of partners, there exists no unifying property shared by all the partners. MLL gene rearrangements are found in leukemias with both lymphoid and myeloid phenotypes and are often associated with infant and secondary leukemias. The immature phenotype of the leukemic blasts suggests an important role for MLL in the early stages of hematopoietic development. Mll homozygous mutant mice are embryonic lethal and exhibit deficiencies in yolk sac hematopoiesis. Recently, two different MLL-containing protein complexes have been isolated. These and other gain- and loss-of-function experiments have provided insight into normal MLL function and altered functions of MLL fusion proteins. This article reviews the progress made toward understanding the function of the wild-type MLL protein. While many advances in understanding this multifaceted protein have been made since its discovery, many challenging questions remain to be answered. PMID- 15779006 TI - An inactive X specific replication origin associated with a matrix attachment region in the human X linked HPRT gene. AB - Early in female mammalian embryogenesis, one of the two X chromosomes is inactivated to compensate the gene dosage between males and females. One of the features of X chromosome inactivation (XCI) is the late replication of the inactivated X chromosome. This study reports the identification, by competitive PCR of nascent DNA, of a replication origin in intron 2 of the human X-linked HPRT gene, that is functional only on the inactive X. Features frequently associated with replication origins, including a peak of enhanced DNA flexibility, a perfect match to the yeast ACS sequence, a 14/15 match to the Drosophila topoisomerase II consensus, and a 20/21 match to an initiation region consensus sequence, were identified close to the replication origin. The origin is located approximately 2 kb upstream of a matrix attachment region (MAR) and also contains two A:T-rich elements, thought to facilitate DNA unwinding. PMID- 15779007 TI - Molecular profile of catabolic versus anabolic treatment regimens of parathyroid hormone (PTH) in rat bone: an analysis by DNA microarray. AB - Teriparatide, human PTH (1-34), a new therapy for osteoporosis, elicits markedly different skeletal responses depending on the treatment regimen. In order to understand potential mechanisms for this dichotomy, the present investigation utilized microarrays to delineate the genes and pathways that are regulated by intermittent (subcutaneous injection of 80 microg/kg/day) and continuous (subcutaneous infusion of 40 microg/kg/day by osmotic mini pump) PTH (1-34) for 1 week in 6-month-old female rats. The effect of each PTH regimen was confirmed by histomorphometric analysis of the proximal tibial metaphysis, and mRNA from the distal femoral metaphysis was analyzed using an Affymetrix microarray. Both PTH paradigms co-regulated 22 genes including known bone formation genes (i.e., collagens, osteocalcin, decorin, and osteonectin) and also uniquely modulated additional genes. Intermittent PTH regulated 19 additional genes while continuous treatment regulated 173 additional genes. This investigation details for the first time the broad profiling of the gene and pathway changes that occur in vivo following treatment of intermittent versus continuous PTH (1-34). These results extend previous observations of gene expression changes and reveal the in vivo regulation of BMP3 and multiple neuronal genes by PTH treatment. PMID- 15779008 TI - TSH receptor protein is selectively expressed in normal human extraocular muscle. AB - Thyroid-associated ophthalmopathy is a common manifestation of Graves' disease. Its pathophysiology is not well understood but an antigen shared between the thyroid and orbit is thought to play a role. Using immunohistochemistry, we have demonstrated the presence of the autoantigenic target of Graves' disease, the thyroid-stimulating hormone receptor, in normal human extraocular muscle. These results support previous findings of the full length and splice variant thyroid stimulating hormone receptor cDNA in extraocular muscle. The observation of the autoimmune target--the thyroid-stimulating hormone receptor protein--being shared between the thyroid and extraocular muscle lends greater support to the notion that an extraocular muscle thyroid-stimulating hormone receptor is also a likely target autoantigen in thyroid-associated ophthalmopathy. PMID- 15779010 TI - Congenital diaphragmatic hernia in WAGR syndrome. AB - Wilms tumor, aniridia, genitourinary anomalies, and mental retardation (WAGR) syndrome is a contiguous gene deletion syndrome involving the Wilms tumor 1 gene (WT1), the paired box gene 6 (PAX6), and possibly other genes on chromosome 11p13. WT1 is required for normal formation of the genitourinary system and the high incidence of Wilms tumor and genitourinary anomalies found in patients with WAGR are attributed to haploinsufficiency of this gene. It has been hypothesized that WT1 also plays an important role in the development of the diaphragm. During mammalian embryonic development, WT1 is expressed in the pleural and abdominal mesothelium that forms part of the diaphragm. Furthermore, mice that are homozygous for a deletion in the mouse homolog of WT1 have diaphragmatic hernias. Case reports describing congenital diaphragmatic hernias in infants with Denys Drash and Frasier syndromes, both of which can be caused by mutations in WT1, provide additional support for this hypothesis. We report an infant with aniridia, bilateral cryptorchidism, vesicoureteral reflux, and a right-sided Morgagni-type diaphragmatic hernia. G-banded chromosome analysis revealed a deletion of 11p12-p15.1. Breakpoint regions were refined by fluorescence in situ hybridization (FISH) and deletion of the WAGR critical region, including WT1, was confirmed. A review of the medical literature identified a second patient with a deletion of 11p13, a left-sided Bochdalek-type diaphragmatic hernia, and anomalies that suggest a diagnosis of WAGR including bilateral microphthalmia, a small penis, bilateral cryptorchidism, and a hypoplastic scrotum. These cases demonstrate that congenital diaphragmatic hernia can be associated with WAGR syndrome and suggest that deletions of WT1 may predispose individuals to develop congenital diaphragmatic hernia. PMID- 15779011 TI - Autosomal recessive mesoaxial synostotic syndactyly with phalangeal reduction maps to chromosome 17p13.3. AB - Previously we have described a novel and distinct form of non-syndromic osseous syndactyly segregating in an autosomal recessive pattern in a consanguineous Pakistani family. The limb findings include mesoaxial reduction of the fingers, synostoses of the third and fourth metacarpals with associated single phalanges, fifth finger clinodactyly, and preaxial webbing of toes. We identified another published report of this phenotype in a large, inbred Turkish family. In the present study we mapped the phenotype in the Pakistani and Turkish families to chromosome 17p13.3 (multipoint LOD score 5.1). The identification of a single locus for this complex limb malformation in two families with distinct ethnic backgrounds supports the hypothesis that this is a distinct form of syndactyly. Since this form of syndactyly is phenotypically distinct from the previously described eight types, we propose to name this phenotype mesoaxial synostotic syndactyly with phalangeal reduction (MSSD, type IX syndactyly, Malik-Percin type). PMID- 15779012 TI - A provisionally unique syndrome of macrosomia, bone overgrowth, macrocephaly, and tall stature. AB - We report a young man with intrauterine macrosomia, macrocephaly, and bony abnormalities. Excessive growth continued throughout infancy and childhood. Bone age was advanced. He developed contractures of the large joints and was confined to a wheelchair. Extensive laboratory studies, repeated on multiple occasions were all normal. Intellectually, he was normal. His near final height was 234 cm. The constellation of findings in this patient is at variance with previously described syndromes of tall stature. We postulate that excessive size and bone overgrowth in this young man is caused by a receptor/post-receptor abnormality involving a growth on/off mechanism at the cellular level. PMID- 15779013 TI - Flaviviruses in motor neuron disease. AB - Sporadic motor neuron disease (MND) causes a progressive loss of motor neurons. West Nile virus can attack motor neurons, so we examined whether flavivirus infection could be detected in MND cases. Spinal cord sections from 22 MND cases were stained immunohistochemically with a flavivirus-specific antibody. No staining for flavivirus was seen in any case. Sporadic MND does not appear to arise from a recent infection with a flavivirus. PMID- 15779014 TI - Yttrium and lanthanide complexes with various P,N ligands in the coordination sphere: synthesis, structure, and polymerization studies. AB - Yttrium and lanthanide complexes with different P,N ligands in the coordination sphere have been synthesized. First the chloride complexes [{CH(PPh2NSiMe3)2}Ln{(Ph2P)2N}Cl] (Ln = Y (1 a), La (1 b), Nd (1 c), Yb (1 d)) having the bulky [CH(PPh2NSiMe3)2]- and the flexible [(Ph2P)2N]- ligands in the same molecule were prepared by three different synthetic pathways. Compounds 1 a d can be obtained by reaction of [{[CH(PPh2NSiMe3)2]LnCl2}2] with [K(thf)nN(PPh2)2] (n = 1.25, 1.5) or by treatment of [{(Ph2P)2N}LnCl2(thf)3] with K[CH(PPh2NSiMe3)2]. Furthermore, a one-pot reaction of K[CH(PPh2NSiMe3)2] with LnCl3 and [K(thf)nN(PPh2)2] leads to the same products. Single-crystal X-ray structures of 1 a-d show that the conformation of the six-membered metallacycle (N1-P1-C1-P2-N2-Ln) which is formed by chelation of the [CH(PPh2NSiMe3)2]- ligand to the lanthanide atom is influenced by the ionic radius of the central metal atom. In solution dynamic behavior of the [(Ph2P)2N]- ligand is observed, which is caused by rapid exchange of the two different phosphorus atoms. Further reaction of 1 b with KNPh2 resulted in [{(Me3SiNPPh2)2CH}La{N(PPh2)2}(NPh2)] (2). Compounds 1 a-d and 2 are active in the ring-opening polymerization of epsilon caprolactone and the polymerization of methyl methacrylate. In some cases high molecular weight polymers with good conversions and narrow polydispersities were obtained. In both polymerizations the catalytic activity depends on the ionic radius of the metal center. PMID- 15779015 TI - The impact of preoperative serum C-reactive protein on the prognosis of patients with hepatocellular carcinoma. AB - BACKGROUND: The authors evaluated the significance of the preoperative serum C reactive protein (CRP) level as a prognostic indicator in patients with hepatocellular carcinoma (HCC). METHODS: One hundred forty-one patients who underwent curative resection for HCC were reviewed retrospectively. Clinicopathologic variables were compared between patients with serum CRP levels >/= 1.0 mg/dL (n = 22 patients; the CRP-positive group) and patients with serum CRP levels < 1.0 mg/dL (n = 119 patients; the CRP-negative group). Univariate and multivariate analyses were conducted to identify factors that affected survival and disease recurrence. RESULTS: There was a significant correlation between the preoperative serum CRP level and tumor size. Invasion to the portal vein in the CRP-positive group was significantly more frequent than that in the CRP-negative group. Even after they underwent curative resection, 75.3% of patients in the CRP positive group experienced recurrence within 1 year. The overall survival and recurrence-free survival rates in the CRP-positive group were significantly lower compared with the rates in the CRP-negative group. On multivariate analysis, the preoperative serum CRP level was selected as one of the unfavorable indicators regarding survival and recurrence. When CRP levels, albumin levels, and platelet counts that were available before surgery were scored as a combined index, the total score demonstrated a good stratification value for survival after hepatic resection. CONCLUSIONS: The current results showed that the preoperative serum CRP level is an independent and significant indicator predictive of poor prognosis and early recurrence in patients with HCC. The new CRP-based scoring system offers reliable information for predicting survival. PMID- 15779016 TI - A cohort study of cancer risk in relation to family histories of cancer in the Utah population database. AB - BACKGROUND: It is well known that genetic variability affects the risk of many cancers, but details of the patterning of inherited cancer risk across different sites and age groups still are not well quantified. METHODS: The authors conducted a nested case-control study of the familial risk of 40 cancers based on a cohort of 662,515 individuals from the Utah Population Database. From 1 to 10 controls selected from the cohort were matched individually on gender, birth year, and birthplace to each cancer case; and familial standardized incidence ratios (FSIR) were calculated for both cases and controls. Conditional logistic regression was used to estimate relative risks and population-attributable risks (PARs) of cancer in relation to FSIR. Relative risks of cancer in first-degree through fifth-degree relatives of cases, compared with controls, were calculated using the proportional hazards methods. All analyses were adjusted for spouse affection status and Latter Day Saints church affiliation. RESULTS: Thirty-five of 40 cancers exhibited positive associations between risk and FSIR, and 21 of those associations were statistically significant. PAR estimates were strikingly high for prostate carcinoma (57%), breast carcinoma (39%), colon carcinoma (32%), lip carcinoma (31%), chronic lymphocytic leukemia (35%), and melanoma (32%). Both the proportion and the number of all cancers attributable to family history peaked at 32% in the group ages 65-84 years and remained high in the group age >/= 85 years. CONCLUSIONS: A substantial portion of cancer risk was attributable to familial factors. The patterns of familial cancer recurrence among distant relatives suggested that simple genetic mechanisms may explain much of the familiality of cancer. PMID- 15779017 TI - Vascular abnormalities in the fingers of patients affected with hereditary hemorrhagic telangiectasia (HHT) as assessed by color doppler sonography. PMID- 15779018 TI - Prevalence of nonsyndromic oral clefts in Texas: 1995-1999. AB - Nonsyndromic cleft lip with/without cleft palate (NSCLP) and nonsyndromic cleft palate only (NSCPO) are common complex birth defects affecting 4,000 newborns annually. We undertook a descriptive study of oral clefts in Texas, focusing on the effect of folic acid fortification and Hispanic ethnicity on the prevalence of oral clefts as these factors have not previously been described. Data on 896 infants with NSCLP and NSCPO born between 1995 and 1999 in Texas were compared to all births in Texas during the same period. Prevalence odds ratios (POR) were calculated for maternal ethnicity, race, age, parity, public health region of residence, highest level of education, and infant gender. The effect of folic acid fortification on oral clefts was also examined. Compared with whites, adjusted POR were 0.97 (95% CI = 0.77-1.23) and 0.90 (95% CI 0.72-1.14) for NSCLP and 0.46 (95% CI = 0.30-0.72) and 0.62 (95% CI = 0.42-0.90) for NSCPO in foreign born and US-born Hispanics, respectively. After fortification was implemented, the rate of NSCLP did not decrease. However, there was a 13% decrease in the prevalence of NSCPO (adjusted POR = 0.87, 95% CI = 0.68-1.15). Compared to whites, the rates in US-born and foreign-born Hispanic women were similar for NSCLP and much lower for NSCPO. The small reduction of 13% in NSCPO after folic acid fortification is imprecise and should be interpreted cautiously. Overall, it appears that folic acid fortification has had very little or no effect on the prevalence of oral clefts in infants born in Texas. PMID- 15779019 TI - Topiramate for weight reduction in Duchenne muscular dystrophy. PMID- 15779020 TI - Peripheral axon caliber and conduction velocity are decreased after burn injury in mice. AB - Peripheral neuropathies are reported to arise as a result of the systemic inflammatory response produced by a full-thickness cutaneous burn injury. This study was designed to characterize the magnitude and time course of functional and morphological changes in peripheral axons that arise after a full-thickness dermal burn injury in an animal model. A 20% body surface area (20% BSA) full thickness dermal burn was applied to the back of C57BL6 female mice. Longitudinal H- and M-wave recordings were used to determine the conduction velocities (CV) of large myelinated motor and sensory axons in the tibial nerve of sham control and burn-injured mice. Motor CVs were significantly reduced from 6 h to 28 days after the burn, and sensory CVs were significantly reduced from 7 to 14 days after the burn. Morphological evaluation also showed that the mean caliber of large axons in tibial nerves and L5 ventral and dorsal roots in burned mice was significantly decreased. The results demonstrate that both functional and morphological deficits may be produced in peripheral nerve axons at sites well removed from a full-thickness dermal burn injury. The neural deficits may contribute to changes in neuromuscular transmission and the development of limb and respiratory muscle weakness that also accompany burn injury. PMID- 15779021 TI - Holoprosencephaly and limb reduction defects: a consideration of Steinfeld syndrome and related conditions. AB - Individuals with holoprosencephaly (HPE) and limb reduction defects have been ascribed historically to a variety of syndromes with overlapping phenotypic features. As such, these patients are challenges for clinicians and researchers alike. In an effort to better understand this association, we reviewed our autopsy records and identified five cases of HPE with reduction defects of the limbs and other anomalies. One case appears to be the third reported instance of Steinfeld syndrome, while others represent microgastria-limb reduction sequence, VATER/VACTERL association, and an additional unique condition characterized by HPE, rhombencephalosynapsis, absent left radius, first metacarpal, and thumb, and congenital heart disease. The phenotypic heterogeneity inherent in these patients continues to complicate diagnosis, which will hopefully be simplified by continuing delineation at morphologic and especially genetic levels. PMID- 15779022 TI - Health-related quality of life during neoadjuvant treatment and surgery for localized esophageal carcinoma. AB - BACKGROUND: Esophagectomy has a negative influence on health-related quality of life (HRQL) during the first postoperative year, but it is not known how chemotherapy or chemoradiotherapy treatment before surgery affects HRQL. The current study examined HRQL during preoperative chemotherapy/chemoradiotherapy treatment and compared postoperative recovery of HRQL in patients undergoing combined treatment with patients undergoing surgery alone. METHODS: One hundred three patients completed standardized HRQL measures before and during neoadjuvant treatment and before and after surgery. Mean HRQL scores were calculated and preoperative scores were used to model postoperative ratings using linear regression. RESULTS: Deterioration in most aspects of HRQL occurred during preoperative chemotherapy. Patients proceeding to concomitant radiotherapy further deteriorated with specific problems with reflux symptoms and role function (difference between means >15, P < 0.01). After neoadjuvant treatment, but before surgery, HRQL returned to baseline levels. Six weeks after surgery, patients reported marked reductions in physical, role, and social function (difference between means > 30, P < 0.01) and increase in fatigue, nausea and emesis, pain, dyspnea, appetite loss, and coughing (difference between means > 15, P < 0.01). Recovery of HRQL was not hampered by preoperative treatment, and fewer problems with postoperative nausea, emesis, and dysphagia were reported by patients who had undergone neoadjuvant treatment compared with patients who had undergone surgery alone. CONCLUSIONS: Preoperative chemotherapy or chemoradiotherapy had a negative impact on HRQL that was restored in patients proceeding to surgery. Recovery of HRQL after esophagectomy was not impaired by neoadjuvant treatment. These results supported the use of neoadjuvant treatment before surgery. PMID- 15779023 TI - Three patients with hallucal polydactyly and WAGR syndrome, including discordant expression of Wilms tumor in MZ twins. AB - The WAGR contiguous gene deletion syndrome is a combination of Wilms tumor, Aniridia, Genito-urinary abnormalities, and growth and mental retardation which is invariably associated with an 11p13 deletion. We report two monozygotic twins and a third, unrelated patient with WAGR syndrome and additional clinical features not usually associated with WAGR. Both twins had developmental delay, growth deficiency, severe ocular involvement (nystagmus, aniridia, cataracts), atrial septal defect and two uncommon findings: agenesis of the corpus callosum and duplication of the halluces. One twin developed Wilms tumors aged 19 months while her sister remained tumor free by the age of 6.5 years. The singleton patient showed typical WAGR syndrome and preaxial hallucal polydactyly. Molecular cytogenetic studies refined the identification of the extent of the deleted segments, which were not identical in the two families. The two deletions included the PAX6 and WT1 genes as previously reported in typical WAGR patients. The unusual anomalies described in this report, may represent the expression of low penetrant traits associated with haploinsufficency of one or more of the genes present in the deletion (PAX6 is expressed in CNS) or may indicate epistatic influences of modifier genes on the expression of gene(s) present in the WAGR region. PMID- 15779024 TI - The correlation between blood oxygenation level-dependent signal strength and latency. AB - PURPOSE: To investigate the relationship between signal strength and latency of the blood oxygenation level-dependent (BOLD) signal. MATERIALS AND METHODS: Several correlation analyses were performed on data obtained in a functional magnetic resonance imaging (fMRI) experiment, where subjects were presented with a simple visual stimulus. The BOLD signal strength was correlated with both the phase shift of the spectral density matrix and time-to-peak calculated from trial averaged time courses. Correlation coefficients were calculated for visual stimuli of 2, 6, and 15 seconds in duration. RESULTS: Analyzing all functional runs for the same subject separately, i.e., including for each run all significantly activated voxels, we observed that correlations between phase shift and signal strength, as well as between time-to-peak and signal strength, decreased with increasing stimulus length. However, when analyses were restricted to voxels found activated in all functional runs, we observed similar correlations between BOLD signal strength and latency in all runs, independent of the length of stimulation. This result was again obtained for both latency measures: the spectral density phase shift and time-to-peak. CONCLUSION: For both latency measures, phase shift and time-to-peak, a high correlation between BOLD signal strength and latency was observed. We have shown that this correlation is independent of the length of visual stimulation. Thus, the correlation between BOLD signal strength and latency seems to be an inherent property of the BOLD response that is independent of the length of stimulation and can be observed using different methods for determining signal latency. PMID- 15779025 TI - In vivo measurements of T1 relaxation times in mouse brain associated with different modes of systemic administration of manganese chloride. AB - PURPOSE: To measure regional T1 and T2 values for normal C57Bl/6 mouse brain and changes in T1 after systemic administration of manganese chloride (MnCl2) at 9.4 T. MATERIALS AND METHODS: C57Bl/6 mice were anesthetized and baseline T1 and T2 measurements obtained prior to measurement of T1 after administration of MnCl2 at 9.4 T. MnCl2 was administered systemically either by the intravenous (IV), intraperitoneal (IP), or subcutaneous (SC) routes. T1 and T2 maps for each MRI transverse slice were generated using commercial software, and T1 and T2 values of white matter (WM), gray matter (GM), pituitary gland, and lateral ventricle were obtained. RESULTS: When compared with baseline values at low-field, significant lengthening of the T1 values was shown at 9.4 T, while no significant change was seen for T2 values. Significant T1 shortening of the normal mouse brain was observed following IV, IP, and SC administration of MnCl2, with IV and IP showing similar acute effects. Significant decreases in T1 values were seen for the pituitary gland and the ventricles 15 minutes after either IV or IP injection. GM showed greater uptake of the contrast agent than WM at 15 and 45 minutes after either IV or IP injections. Although both structures are within the blood-brain barrier (BBB), GM and WM revealed a steady decrease in T1 values at 24 and 72 hours after MnCl2 injection regardless of the route of administration. CONCLUSION: Systemic administration of MnCl2 by IV and IP routes induced similar time-course of T1 changes in different regions of the mouse brain. Acute effects of MnCl2 administration were mainly influenced by either the presence or absence of BBB. SC injection also provided significant T1 change at subacute stage after MnCl2 administration. PMID- 15779026 TI - Parallel acquisition techniques for accelerated volumetric interpolated breath hold examination magnetic resonance imaging of the upper abdomen: assessment of image quality and lesion conspicuity. AB - PURPOSE: To evaluate the impact of parallel acquisition techniques (PATs) on image quality and detection of liver metastases using three-dimensional volumetric interpolated breath-hold examination (VIBE) for clinical liver imaging. MATERIALS AND METHODS: Forty-nine patients with various primary malignancies underwent abdominal dynamic contrast-enhanced three-dimensional VIBE magnetic resonance imaging (MRI) (1.5 T) using a standard phased array coil. Recently introduced Generalized Autocalibrating Partially Parallel Acquisition (GRAPPA) and SENSitivity Encoding (mSENSE) PAT reconstruction algorithms were added to reduce scan time twofold. Overall image quality, motion, and aliasing artifacts were classified on a 5-point scale. Signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) measurements were performed for quantitative comparison. All sequences were evaluated concerning the number of detected lesions. RESULTS: PAT resulted in a reduction of data acquisition time from 23 to 13 seconds. Both GRAPPA and mSENSE data sets yielded 30% less SNR (34.8 +/- 14.1 and 33.1 +/- 13.3, P < 0.001) and 35% less CNR (21.2 +/- 15.0 and 20.9 +/- 12.7, P < 0.05) in comparison to unaccelerated VIBE (SNR = 50.8 +/- 20.3/CNR = 32.5 +/- 19.1). Similarly, PAT revealed lower-image-quality scores than unaccelerated VIBE. GRAPPA resulted in more fold-over artifacts than mSENSE. mSENSE revealed slightly fewer motion artifacts than no PAT. The unaccelerated late-venous-phase VIBE sequence revealed 146 lesions in the same patients. Accelerated images with mSENSE reconstruction detected 138 lesions. GRAPPA revealed 127 lesions, and thus performed inferior to mSENSE. CONCLUSION: At least for arrays with small numbers of elements, such as arrays used in this study, the PAT-induced reduction in scanning times must be weighed against compromises in image quality, which translate into poorer diagnostic performance regarding detection of small hepatic lesions. Thus, the PAT implementations tested in this study should probably be reserved for patients unable to hold their breaths for regular three-dimensional VIBE data sets. PMID- 15779027 TI - Magnetic resonance image-guided trans-septal puncture in a swine heart. AB - PURPOSE: To test the feasibility of performing magnetic resonance (MR)-guided trans-septal punctures in the swine heart. MATERIALS AND METHODS: All procedures were performed in a 1.5-T MR scanner. A novel, active MR intravascular needle system was utilized for needle tracking and septal punctures. Trans-septal punctures were performed in five swine using electrocardiogram (ECG)-gated high resolution and non-ECG-gated, real-time MR imaging techniques. The intravascular needle was advanced over a guidewire from the femoral vein. Once the needle was in proper position, trans-septal punctures were made. RESULTS: Active tracking of the needle traversing the septum was possible. The location of the catheter tip was confirmed using real time gradient recalled echo (GRE). After a confirmatory ventriculogram with gadolinium-DTPA, a 0.014-inch guidewire was advanced into the left atrium and left ventricle. All punctures were made with no change in cardiac rhythm or rate; postmortem analysis was performed on all animals and demonstrated that 18/19 (95%) punctures were directly through the fossa ovalis. CONCLUSION: Using only MR guidance and a novel active intravascular needle system, we were able to repeatedly puncture the fossa ovalis in a swine heart from a transfemoral approach, with direct visualization of all components, including the needle, the atria, the fossa ovalis, and the surrounding vasculature. PMID- 15779028 TI - Dobutamine-induced stress affects intracellular uptake of manganese: a quantitative magnetic resonance imaging study in pigs. AB - PURPOSE: To determine whether there are differences in increase in longitudinal relaxation rate (delta R1) in the myocardium between bolus administration of Mn2+ ions during rest and during dobutamine-induced stress and, additionally, to determine whether there are differences in delta R1 between bolus injection and infusion of Mn2+ ions during dobutamine-induced stress. MATERIALS AND METHODS: Pigs were divided into three groups with six pigs in each group. All animals received 15 micromol MnCl2/kg of body weight (b.w.) intravenously either as a bolus injection (groups 1 and 2) or as an infusion over one minute (group 3). The animals in groups 2 and 3 were subjected to dobutamine stress before injection of MnCl2, while those in group 1 were not given dobutamine. T1 was quantified in the myocardium and left ventricular blood pool before contrast injection and repeatedly during a one-hour postinjection period. RESULTS: A significant difference in delta R1 between the groups with and the group without dobutamine stress was noted in the myocardium up to 45 minutes after contrast agent injection. No such significant difference was found between pigs that received the contrast agent as a bolus injection compared to infusion. CONCLUSION: Dobutamine stress increases uptake of manganese ions in the myocardium. This increase was independent of whether the contrast agent was administered as a bolus injection or as an infusion. PMID- 15779029 TI - Small bowel motility assessment with magnetic resonance imaging. AB - PURPOSE: To assess primarily the feasibility of magnetic resonance imaging (MRI) as a tool to monitor small bowel peristaltic motion and secondarily to validate this technique by demonstrating drug-induced motility changes. MATERIALS AND METHODS: After a standardized oral preparation of Ispaghula husk (Metamucil) and meglumine gadoterate (Gd-DOTA; Dotarem), 10 volunteers underwent dynamic MRI using a two-dimensional turbofast field echo (TFE) sequence with a slice repetition time of 500 msec. Intraluminal cross-sectional caliber changes over time were assessed allowing quantification of the peristaltic frequencies and amplitudes of the small gut on various regions of interest. Pharmacologically induced alterations of the peristaltic motion after spasmolytics and gastrokinetic motion enhancers were investigated. RESULTS: Small bowel diameter measurement resulted in a peristalsis of 10.96 (SD = +/-2.51) waves per minute, oscillating regularly with mean amplitudes of 6.65 mm (SD = +/-1.15 mm). Peristaltic frequency in normal individuals is consistent with that observed with other techniques. Intravenous administration of scopolamine butylbromide (= hyoscine butylbromide/Buscopan) resulted in small bowel paralysis within 21.3 seconds (SD = +/-2.8 seconds). Prokinetic effect of intravenous metoclopramide (Paspertin) after Buscopan paralysis was tested in one volunteer, characterized by a slow recovery of peristalsis, which propagated from the proximal to the distal segments and enhanced contraction amplitudes. CONCLUSION: Dynamic MRI allows observing and quantifying small bowel peristalsis, characterizing motion patterns, and monitoring the effects of interfering factors such as drugs. PMID- 15779030 TI - High-resolution diffusion-weighted imaging with interleaved variable-density spiral acquisitions. AB - PURPOSE: To develop a multishot magnetic resonance imaging (MRI) pulse sequence and reconstruction algorithm for diffusion-weighted imaging (DWI) in the brain with submillimeter in-plane resolution. MATERIALS AND METHODS: A self-navigated multishot acquisition technique based on variable-density spiral k-space trajectory design was implemented on clinical MRI scanners. The image reconstruction algorithm takes advantage of the oversampling of the center k space and uses the densely sampled central portion of the k-space data for both imaging reconstruction and motion correction. The developed DWI technique was tested in an agar gel phantom and three healthy volunteers. RESULTS: Motions result in phase and k-space shifts in the DWI data acquired using multishot spiral acquisitions. With the two-dimensional self-navigator correction, diffusion-weighted images with a resolution of 0.9 x 0.9 x 3 mm3 were successfully obtained using different interleaves ranging from 8-32. The measured apparent diffusion coefficient (ADC) in the homogenous gel phantom was (1.66 +/- 0.09) x 10(-3) mm2/second, which was the same as measured with single-shot methods. The intersubject average ADC from the brain parenchyma of normal adults was (0.91 +/- 0.01) x 10(-3) mm2/second, which was in a good agreement with the reported literature values. CONCLUSION: The self-navigated multishot variable density spiral acquisition provides a time-efficient approach to acquire high resolution diffusion-weighted images on a clinical scanner. The reconstruction algorithm based on motion correction in the k-space data is robust, and measured ADC values are accurate and reproducible. PMID- 15779031 TI - Driven equilibrium magnetic resonance imaging of articular cartilage: initial clinical experience. AB - PURPOSE: To evaluate three-dimensional driven equilibrium Fourier transform (3D DEFT) for image quality and detection of articular cartilage lesions in the knee. MATERIALS AND METHODS: We imaged 104 consecutive patients with knee pain with 3D DEFT and proton density (PD-FSE) and T2-weighted (T2-FSE) fast spin echo. Twenty four went on to arthroscopy. Signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) efficiency were measured. Subjective image quality, fat suppression, and cartilage thickness visibility were assessed. Cartilage lesions on 3D-DEFT and T2-FSE were compared with findings outlined in operative reports. RESULTS: SNR efficiency was higher for 3D-DEFT and PD-FSE than for T2-FSE (P < 0.02). 3D DEFT and PD-FSE showed superior cartilage thickness visibility compared with T2 FSE (P < 0.02). T2-FSE showed better fat suppression and fewer image artifacts than 3D-DEFT (P < 0.04). 3D-DEFT had similar sensitivity and similar specificity for cartilage lesions compared with PD-FSE and T2-FSE. CONCLUSION: 3D-DEFT provides excellent synovial fluid-to-cartilage contrast while preserving signal from cartilage, giving this method a high cartilage SNR. 3D-DEFT shows the full cartilage thickness better than T2-FSE. T2-FSE had superior fat saturation and fewer artifacts than 3D-DEFT. Overall, 3D-DEFT requires further technical development, but is a promising method for imaging articular cartilage. PMID- 15779032 TI - Combined helium-3/proton magnetic resonance imaging measurement of ventilated lung volumes in smokers compared to never-smokers. AB - PURPOSE: To use a combination of helium-3 (3-He) magnetic resonance imaging (MRI) and proton single-shot fast spin echo (SSFSE) to compare ventilated lung volumes in groups of "healthy" smokers, smokers diagnosed with moderate chronic obstructive pulmonary disease (COPD), and never-smokers. MATERIALS AND METHODS: All study participants were assessed with spirometry prior to imaging. 3-He images were collected during an arrested breath hold, after inhaling a mixture of 200 mL of hyperpolarized 3-He/800 mL of N2. Proton SSFSE images were acquired after inhaling 1 liter of room air. The ventilated volume for each study participant was calculated from the 3-He images, and a ratio was calculated to give a percentage ventilated lung volume. RESULTS: Never-smokers exhibited a 90% mean ventilated volume. The mean ventilated lung volumes for healthy smokers and smokers diagnosed with COPD were 75.2% and 67.6%, respectively. No correlation with spirometry was demonstrated for either of the smoking groups. CONCLUSION: Combined 3-He/Proton SSFSE MRI of the lungs is a noninvasive method, using nonionizing radiation, which demonstrates ventilated airspaces and enables the calculation of ventilated lung volumes. This method appears to be sensitive to early obstructive changes in the lungs of smokers. PMID- 15779033 TI - Differential uptake of ferumoxtran-10 and ferumoxytol, ultrasmall superparamagnetic iron oxide contrast agents in rabbit: critical determinants of atherosclerotic plaque labeling. AB - PURPOSE: To compare atherosclerotic plaque uptake of a first (ferumoxtran-10) and second generation (ferumoxytol) ultrasmall superparamagnetic iron oxide (USPIO) contrast agent with different pharmacokinetic/pharmacodynamic properties. MATERIALS AND METHODS: New Zealand White rabbits maintained on a high cholesterol/fat diet were subjected to balloon injury to the abdominal aorta. Ferumoxtran-10 or ferumoxytol (500 micromol/kg) was administered at 2, 4, and 8 weeks following injury. In vivo magnetic resonance imaging (MRI) was performed immediately prior to, immediately after, and 6 days post-contrast administration. Ex vivo MRI, histologic, and inductively coupled plasma-mass spectrometry (ICP MS) iron analyses were performed on the excised vessels. RESULTS: The blood pool clearance of ferumoxytol (t(1/2) < or = 6 hours) was more rapid than that of ferumoxtran-10 (t(1/2) < or = 48 hours). Decreased in vivo MRI signal intensity in the abdominal aorta was observed at 2, 4, and 8 weeks following injury with ferumoxtran-10, but not with ferumoxytol. Consistent with these observations, ex vivo MRI signal intensity was decreased in the ferumoxtran-10 vessels, and to a lesser degree in the ferumoxytol vs. control vessels (- contrast agent). In contrast, in vitro macrophage phagocytosis of USPIO was four to six fold greater with ferumoxytol than with ferumoxtran-10. Additionally, the absolute iron content correlated with ex vivo MRI signal intensity in all vessels (r = -0.86, P < 0.0001). CONCLUSIONS: These data suggest that the exposure period of atherosclerotic plaque to USPIO rather than the kinetics of the USPIO uptake by plaque alone is a critical criterion for experimental design of in vivo studies. PMID- 15779034 TI - A multicenter study of two magnetic resonance spectroscopy techniques in individuals with HIV dementia. AB - PURPOSE: To evaluate single-voxel proton magnetic resonance spectroscopy (SV-MRS) and magnetic resonance spectroscopic imaging (MRSI) metabolite results in individuals with HIV dementia. MATERIALS AND METHODS: Twenty HIV-positive (HIV+) individuals underwent SV-MRS (TE 35 msec) and MRSI (TE 280 msec). Results were stratified according to serostatus, dementia severity, psychomotor speed performance, and functional impairment. RESULTS: HIV+ individuals with psychomotor slowing had an increased myoinositol/creatine (mI/Cr) ratio (0.63 vs. 0.45) in the frontal white matter using SV-MRS and an increased choline (Cho)/Cr ratio (1.88 vs. 1.41) in the mesial frontal gray matter using MRSI compared to HIV+ individuals without psychomotor slowing. Using MRSI, subjects with HIV dementia also had a decreased N-acetyl aspartate (NAA)/Cho ratio (1.55 vs. 2.53) compared to HIV+ individuals without cognitive impairment in the mesial frontal gray matter. Both techniques detected metabolite ratio abnormalities associated with abnormal functional performance. CONCLUSION: SV-MRS and MRSI offer complementary roles in evaluating individuals with HIV dementia. Short TE SV-MRS measures mI, which may be elevated in early HIV dementia, whereas MRSI provides wider spatial coverage to examine specific regional changes. PMID- 15779035 TI - Optimization of the arterial input function for myocardial perfusion cardiovascular magnetic resonance. AB - PURPOSE: To determine how injection rate, cardiac function, and breathhold influence the arterial input function (AIF), in order to optimize the AIF in the clinical setting for quantitative myocardial perfusion cardiovascular magnetic resonance (CMR). MATERIALS AND METHODS: Gd (0.1 mmol/kg) bolus was injected at 3, 5, or 7 mL/second in 35 patients. In each cardiac cycle during the first-pass, a series of saturation recovery (SR) fast low-angle shot (FLASH) low resolution images with exponentially increasing SR delay times were acquired. Signal intensity (SI) time measurements were made from a region of interest (ROI) drawn in the ascending aorta (AA). The calculation of short T1s and thus peak Gd concentration [Gd] was performed by fitting the mean ROI SI against SR delay times. RESULTS: The mean peak [Gd] in the AA increased as injection rate increased from 3 mL/second (5.0 mM), to 5 mL/second (7.1 mM), to 7 mL/second (4 mM) (P < 0.0001). The peak [Gd] increased as the left ventricular stroke volume (LV SV) increased (P = 0.01). Breath holding was not found to influence peak [Gd]. CONCLUSION: In this study, we found that a high injection rate has advantages over lower injection speeds, although the duration of the AIF was apparently not significantly shortened by faster injection. The choice of expiration or inspiration as breathhold did not have a significant influence upon the AIF. Poor cardiac function was associated with a lower peak [Gd], indicating that first pass perfusion measurements in these patients will be suboptimal. PMID- 15779036 TI - Improved artery delineation in dual-stack coronary magnetic resonance angiography using parallel imaging at 3 T. AB - PURPOSE: To improve vessel sharpness and T2 preparation (T2Prep) in dual-stack three-dimensional coronary magnetic resonance angiography (MRA) by shortening the time delays between the magnetization preparation pulses and the imaging stacks using sensitivity encoding (SENSE) at 3 T. MATERIALS AND METHODS: By combining dual-stack three-dimensional coronary MRA with the parallel imaging technique SENSE at 3 T, the acquisition duration of each three-dimensional imaging stack was shortened by a factor of 2. The proposed technique was implemented and tested in experiments with a moving phantom and in measurements on six healthy volunteers. RESULTS: The time delay between the navigator, T2Prep, and second imaging stack was reduced by 37%, relative to conventional dual-stack angiography without parallel imaging. This enabled the achievement of comparable high-vessel sharpness values for the left and the right coronary arteries relative to values known from conventional single-stack three-dimensional coronary MRA at 3 T. CONCLUSION: Parallel imaging allows for improved vessel visualization in dual stack coronary MRA, given shorter temporal delays between navigator, T2Prep, and the actual image acquisitions, and thus considerably facilitates simultaneous acquisition of high-resolution angiograms of the left and right coronary systems. PMID- 15779037 TI - Reproducibility and dependence on diffusion weighting of line scan diffusion in the lumbar intervertebral discs. AB - PURPOSE: To investigate the dependence of line scan diffusion imaging (LSDI) in the lumbar vertebral discs on diffusion weighting, fat suppression (FS), and postprocessing noise correction. MATERIALS AND METHODS: Eleven normal volunteers were scanned using 4 b-value and 12 b-value LSDI protocols, with and without FS. Three repeated four b-value scans were performed for evaluation of the reproducibility of apparent diffusion coefficient (ADC) values calculated with mono- and biexponential decay models. Two-point ADC analysis for 12 b-value scans was performed with and without noise correction to evaluate the ADC dependence on diffusion weighting. Correlations between different ADC calculation and acquisition methods were evaluated. RESULTS: Monoexponential ADC measures had a coefficient of variation (CV) under 3%, while use of a constrained biexponential increased the CV to 6% to 9%. Strong dependence on b-value was seen from chemically shifted marrow fat signal and noise. These systematic variations in ADC were eliminated using noise correction and FS. ADC values from 4 and 12 b value FS scans correlated strongly (R2 = 0.91), while biexponentially derived ADC values correlated moderately well with the FS ADC (R2 = 0.51). CONCLUSION: LSDI gives reproducible ADC measurements in the lumbar discs, largely independent of b value and signal-to-noise ratio (SNR) when used with noise correction and FS. PMID- 15779038 TI - Magnetic resonance voiding cystography in the diagnosis of vesicoureteral reflux: comparative study with voiding cystourethrography. AB - PURPOSE: To evaluate the feasibility of magnetic resonance voiding cystography (MRVC) compared with voiding cystourethrography (VCUG) for detecting and grading vesicoureteral reflux (VUR). MATERIALS AND METHODS: MRVC was performed upon 20 children referred for investigation of reflux. Either coronal T1-weighted spin echo (SE) or gradient-echo (GE) (fast multiplanar spoiled gradient-echo (FMPSPGR) or turbo fast low-angle-shot (FLASH)) images were obtained before and after transurethral administration of gadolinium solution, and immediately after voiding. The findings of MRVC were compared with those of VCUG and technetium-99m ((99m)Tc) dimercaptosuccinic acid (DMSA) single-photon emission computed tomography (SPECT) performed within 6 months of MRVC. RESULTS: VUR was detected in 23 ureterorenal units (16 VURs by both methods, 5 VURs by VCUG, and 2 VURs by MRVC). With VCUG as the standard of reference, the sensitivity of MRVC was 76.2%; the specificity, 90.0%; the positive predictive value, 88.9%; and the negative predictive value, 78.3%. There was concordance between two methods regarding the grade of reflux in all 16 ureterorenal units with VUR detected by both methods. Of 40 kidneys, MRVC detected findings of renal damage or reflux nephropathy in 13 kidneys, and (99m)Tc DMSA renal SPECT detected findings of reflux nephropathy in 17 kidneys. CONCLUSION: Although MRVC is shown to have less sensitivity for VUR than VCUG, MRVC may represent a method of choice offering a safer nonradiation test that can additionally evaluate the kidneys for changes related to reflux nephropathy. PMID- 15779039 TI - Magnetic resonance angiography of the carotid arteries using three different techniques: accuracy compared with intraarterial x-ray angiography and endarterectomy specimens. AB - PURPOSE: To compare three different magnetic resonance angiography (MRA) techniques with x-ray angiography and endarterectomy specimens. MATERIALS AND METHODS: Twenty-one patients underwent x-ray angiography, three-dimensional time of-flight (TOF) focusing on the carotid bifurcation, high-resolution (HR) contrast-enhanced (CE) MRA, and time-resolved CE MRA. Stenoses of internal carotid arteries were evaluated by three independent observers on identical projection of x-ray angiography and MRA. Maximum stenosis grades on MRA were assessed additionally and correlated with endarterectomy specimens in 12 cases. RESULTS: Sensitivity for the detection of severe stenoses was excellent (100%) for all MRA techniques, and specificity was superior for three-dimensional TOF (96.7%) compared with HR CE MRA (80.6%) and time-resolved CE MRA (83.9%). The correlation between x-ray angiography and MRA for all stenoses was slightly superior for three-dimensional TOF and HR CE MRA compared with the time-resolved technique (kappa = 0.87 and 0.86 vs. 0.84). The same trend was seen for the interobserver agreement and for the correlation with endarterectomy specimens. Eleven up to 17 stenoses (depending on the MRA technique) were graded higher using additional projections. CONCLUSION: Three-dimensional TOF MRA yielded even more accurate results than HR CE MRA in grading of stenoses near the carotid bifurcation. Therefore, a combination of both methods seems to be advantageous. PMID- 15779040 TI - Effect of ingestion order of the fat component of a solid meal on intragastric fat distribution and gastric emptying assessed by MRI. AB - PURPOSE: To develop an MRI technique to investigate how varying the ingestion order of nonfat and fat components of a solid meal influences three-dimensional intragastric distribution and gastric emptying (GE). MATERIALS AND METHODS: Eight healthy subjects were studied twice in randomized order. On one occasion (condition F-NF), the fat component (40 g mayonnaise on toast) was served before the nonfat component (270 g pasta, 200 g tomato sauce, 100 mL water); on the other (condition NF-F), the ingestion order was reversed. GE and intragastric distribution of both components were assessed by MRI for 180 minutes. RESULTS: During condition F-NF, GE of fat was significantly faster than during condition NF-F (T(25) [min]: F-NF: 20 +/- 9; NF-F: 40 +/- 7; P < 0.05), a larger amount of fat was observed in the antrum during condition F-NF, and more fat layering occurred. No differences were observed in total GE between the two conditions. CONCLUSION: Meal ingestion order influences GE and intragastric distribution of fat, which can be assessed by MRI techniques, providing new insights into the physiology of gastric processing and intragastric distribution of different meal phases. PMID- 15779041 TI - Adding another spectral dimension to 1H magnetic resonance spectroscopy of hepatic encephalopathy. AB - PURPOSE: To evaluate a localized two-dimensional correlated magnetic resonance spectroscopic (L-COSY) technique in patients with hepatic encephalopathy (HE) and healthy subjects, and to correlate the cerebral metabolite changes with neuropsychological (NP) test scores. MATERIALS AND METHODS: Eighteen minimal hepatic encephalopathy (MHE) patients and 21 healthy controls have been investigated. A GE 1.5-T magnetic resonance (MR) scanner was used in combination with a body MR coil for transmission and a 3-inch surface coil for reception. A 27-mL voxel was localized by three slice-selective radio frequency (RF) pulses (90 degrees-180 degrees-90 degrees) in the anterior cingulate region. The total duration of each two-dimensional L-COSY spectrum was approximately 25 minutes. The NP battery included a total of 15 tests, which were grouped into six domains. RESULTS: MR spectroscopic results showed a statistically significant decrease in myo-inositol (mI) and choline (Ch) and an increase in glutamate/glutamine (Glx) in patients when compared to healthy controls. There was also an increase in taurine (Tau) in patients. The NP results indicated a significant correlation between motor function assessed by NP tests and mI ratios recorded using two dimensional L-COSY. CONCLUSION: The study demonstrated the feasibility of evaluating the two-dimensional L-COSY sequence in a clinical environment. The results showed additional cerebral metabolites that can be measured with the technique in comparison to one-dimensional study. PMID- 15779042 TI - Cell proliferation in the developing lateral line system of zebrafish embryos. AB - The sensory organs of the embryonic lateral line system are deposited by migrating primordia that originate in the otic region. Here, we examine the pattern of cell proliferation in the posterior lateral line system. We conclude that three phases of cell proliferation are involved in the generation of this system, separated by two phases of mitotic quiescence. The first phase corresponds to generalized proliferation during gastrulation, followed by a first period of quiescence that may be related to the determination of the lateral line precursor cells. A second phase of proliferation takes place in the placode and migrating primordium. This region is organized in annuli that correspond to the expression of proneural/neurogenic genes. A second period of quiescence follows, corresponding to deposition and differentiation of the sensory organs. The third period of proliferation corresponds to continued renewal of hair cells by division of support cells within each sensory organ. PMID- 15779043 TI - Structure of YciI from Haemophilus influenzae (HI0828) reveals a ferredoxin-like alpha/beta-fold with a histidine/aspartate centered catalytic site. PMID- 15779044 TI - Pathogenic potentials of glycoprotein C-negative syncytial mutants from rabbit T cells infected persistently with herpes simplex virus type 1. AB - Human T cell lymphotropic virus type I (HTLV-I)-transformed T cells of rabbits were infected persistently with Herpes simplex virus type 1 (HSV-1) strain KOS. These infected cells yielded syncytial mutants, either glycoprotein C (gC) negative or -positive, which predominated over and replaced the wild-type virus in a long-term culture for 2 years. An alignment of nucleotide sequences showed multiple mutations in glycoprotein B (gB) and gC genes of these mutants, which are or may be responsible for the mutant phenotypes. One of four mutants analyzed produced extensively large syncytia and possessed point mutations within the cytoplasmic domain of gB. All four mutants possessed multiple point mutations in gC and two possessed single insertions which resulted in a frame shift, leading to the premature termination of the gC polypeptide chain. The supernatant of the 2-year culture of cells infected persistently, containing only gC-negative syncytial mutants, induced encephalitic symptoms in B/Jas inbred rabbits, when injected intravenously. One gC-negative syncytial isolate from an encephalitic lesion, together with those from the culture supernatant, were examined for pathogenic potential in vitro and in vivo. All these mutants were more cytotoxic and more susceptible to complement inactivation than the parental virus, and could infect and replicate in adrenal glands when injected intravenously into rabbits. Invasion into the central nervous system appeared to be blocked at the portal of entry, the adrenal gland, i.e., none exhibited neuroinvasive potential by itself. Syncytial gC-negative mutants could thus be pathogenic in rabbits. PMID- 15779045 TI - Detection of serum antibodies to bovine norovirus in veterinarians and the general population in the Netherlands. AB - The close genetic relationship of human and animal strains of norovirus has raised the possibility of transmission of noroviruses from animals to humans and may explain the emergence of certain norovirus strains. To assess if exposure to bovine noroviruses (NoV) might result in infection in humans, an enzyme immunoassay (EIA) was designed and validated in order to detect antibodies against bovine norovirus. This and two other EIAs were used to test sera from 210 veterinarians and 630 matched population controls for IgG and IgA antibodies to recombinant capsid protein of bovine NoV (rBoV), Norwalk virus (rNV), and Lordsdale virus (rLDV). Of 840 participants, IgG reactivity to rBoV was found in 185 (22%), to rNV in 638 (76%) and to rLDV in 760 (90%). IgG reactivity to rBoV was more common in veterinarians (58/210: 28%) than in controls (127/630: 20% [P = 0.03]). IgA reactivity to rBoV was similar in both veterinarians and controls. Cross-reactivity of IgA and IgG antibodies to rBoV and rNV was seen, but 26% of all specimens positive rBoV antibodies showed high IgG reactivity to rBoV but low reactivity to rNV, suggesting a specific response to bovine antigen. No evidence of overall cross-reactivity of antibodies to rBoV and rLDV was seen. Among veterinarians, youth spent on farm (Odds Ratio [OR] = 1.8) and membership of the bovine practitioners' society (OR = 2.7) were significantly associated with IgG seroreactivity to rBoV. These data indicate that bovine strains of NoV may infect humans though less frequently than human strains. PMID- 15779046 TI - Emerging patterns of cardiac conduction in the chick embryo: waveform analysis with photodiode array-based optical imaging. AB - Major difficulties investigating the developing cardiac conduction system stem from that the embryonic heart is extremely small (< 2 mm) and cardiac activation is relatively rapid (< 8 msec). The objective of this study was to investigate the electrophysiology of the embryonic chick cardiac conduction system at periseptation stages with a photodiode array-based detection method of optical mapping capable of high spatial and temporal resolution. Previous work indicated that, in chicken embryos, a switch occurs in ventricular activation pattern from immature base-to-apex to mature apex-to-base pattern at the time of ventricular septation. It was our aim to map activation in more detail to identify the active pathway or pathways of atrioventricular conduction at these particular stages. Analysis of preseptated hearts (n = 10) showed that the latest atrial activation took place just above the site of the earliest ventricular activation at the ventral left ventricular base. Analysis of postseptated hearts (n = 11) showed apex-to-base conduction consistent with activation through the maturing His Purkinje system. Evaluation of hearts during septation revealed a gradual transition of ventricular activation patterns rather than an abrupt "switch." External pacing of preseptated hearts revealed significant slowing of interventricular conduction compared with spontaneous beats (spontaneous, 61.7 cm/sec +/- 9 cm/sec vs. paced, 36.5 cm/sec +/- 10 cm/sec). The more detailed mapping revealed that, before septation, the pattern of activation of the ventricular myocardium is consistent with direct atrial-ventricular myocardial connections at the left lateral atrioventricular junction; however, functional evidence for a preferential conduction pathway within the ventricles was present before septation. PMID- 15779047 TI - Establishment of mesodermal gene expression patterns in early Xenopus embryos: the role of repression. AB - In Xenopus, activin-like signals are able to induce and pattern mesoderm in a concentration-dependent manner. Previous experiments demonstrated that discrete gene expression patterns can be formed in animal cap explants as a response to graded activin signals. We analyzed the spatiotemporal appearance of goosecoid (gsc), chordin (chd), and Xbrachyury (Xbra) mRNAs in whole Xenopus embryos ectopically expressing activin or BVg1. To discriminate between direct transcriptional regulation and indirect, protein synthesis-dependent effects of ectopic signals, we combined overexpression studies and cycloheximide treatment. Our experiments revealed long-range signaling of activin/BVg1, but the expression patterns of gsc, chd, and Xbra in response to activin/BVg1 indicated that repressors are essential to establish the proper expression of these genes. Analysis of endogenous gsc, chd, and Xbra transcript distribution in embryos treated with cycloheximide supported this concept. We, therefore, conclude that inhibition is fundamental during early embryonic patterning. PMID- 15779048 TI - Persistence of acute infection with hepatitis B virus genotype A and treatment in Japan. AB - Among the 97 adult patients with acute hepatitis B who were admitted to the Toranomon Hospital in Metropolitan Tokyo during 28 years from 1976 to 2003, 31 (32%) were infected with hepatitis B virus (HBV) genotype A, nine (9%) with genotype B, 44 (45%) with genotype C, one (1%) each with genotypes E and F. HBV in the remaining 11 (11%) patients were untypeable. All the 31 patients with acute hepatitis B caused by HBV genotype A infection were male with a median age of 31 years, and 16 (52%) contracted infection through extramarital sexual contacts. The baseline HBV DNA level was higher in the seven (23%) patients in whom infection with HBV genotype A persisted than the remaining 24 (77%) with spontaneous resolution (median: >8.7 vs. 6.0 log genome equivalents/ml, P = 0.004). Persistent infection was more frequent in patients with maximum alanine aminotransferase <500 IU/L than > or =500 IU/L (83% [5/6] vs. 4% [1/25], P = 0.0001). Of the six patients with persistent HBV genotype A infection who received interferon and/or lamivuidine for treatment of chronic active hepatitis, three (50%) responded with the loss of hepatitis B e antigen (HBeAg); hepatitis B surface antigen (HBsAg) was cleared from serum in one patient who received interferon and lamivudine in sequence. HBV genotype A persisted along with HBeAg in the remaining three patients given antiviral therapy as well as another who was not treated. In conclusion, infection with HBV genotype A prevails in patients with acute hepatitis B in Japan where genotypes B and C are common, is often contracted sexually (16/31 [52%]) and tends to persist (7/31 [23%]). Infection was cleared in only one of the six (17%) patients who received antiviral therapy. PMID- 15779049 TI - Prevalence of antibodies to hepatitis E virus in residents of a district in Havana, Cuba. AB - A seroepidemiological study of hepatitis E virus (HEV) infection was conducted in a district of Havana, where hepatitis A virus (HAV) is considered endemic. The levels of anti-HEV antibodies were evaluated by enzyme-linked immunosorbent assay (ELISA) based on the recombinant protein GST-ORF2.1. Anti-HEV antibodies were detected in 11 of 209 (5.3%) of serum samples, compared to 71.3% for anti-HAV antibodies. No risk factors reported previously for HEV infection showed a significant association with the presence of anti-HEV antibodies, whereas anti HAV antibodies were strongly associated with increasing age. HEV may be considered endemic in this area and is likely to have a significant clinical impact. PMID- 15779051 TI - Contribution of morphometry in the differential diagnosis of fine-needle thyroid aspirates. AB - BACKGROUND: Cytologic discrimination of cellular nodules, follicular adenoma, and follicular carcinoma in the thyroid is problematic. Methods are needed to achieve a reliable diagnosis. Some sophisticated tools, such as microarrays, offer great potential but lack accompanying morphologic information. METHODS: One hundred twelve samples obtained from patients with lesions histopathologically diagnosed as nodular goiter, follicular adenoma, follicular carcinoma, and papillary carcinoma were used. Eight geometric features, such as nuclear area and circular form factor, were measured. The dataset was divided into six overlapping groups to represent the frequently encountered situations in routine practice. Multivariate analysis of variance, Tukey's honestly significant differences test, and discriminant analysis were performed. Statistical analysis was carried out with two conceptually different approaches. In the first, data from all measured nuclei were used. In the second, a subset of data representing the most extreme values of variables was extracted from the entire dataset to simulate the "selection procedure" performed during conventional morphologic examination. RESULTS: When the selected dataset instead of data from all measured nuclei was used, the correct classification rates in discriminant analysis improved considerably. CONCLUSIONS: Morphologic examination is based primarily on selection. Using data obtained from all of the cells in morphometry may cause a dilution effect in diagnostically important features. Morphometric studies may also be planned with a proper selection "bias." This may be particularly helpful when isolated abnormal cells carry most of the diagnostic information. PMID- 15779052 TI - Coping with appraised breast cancer risk among women with family histories of breast cancer. AB - This descriptive correlational study was based on Neuman's Systems Model and was designed to examine how women with family histories of breast cancer appraise and cope with breast cancer risk. Ninety percent of 209 women in the sample appraised their degree of breast cancer threat as moderate or high. Women with high degrees of appraised risk had low breast cancer risk scores, while women with moderate degrees of appraised risk had higher risk scores. The most common and effective coping modes used were confrontive, optimistic, and early detection behaviors. Over 75% of women either did not use evasive, emotive, palliative, and fatalistic modes of coping, or rated them as ineffective. Canonical correlation analyses revealed five different patterns of appraisal and coping behaviors and lent support to the premise that the type of coping behaviors used varies with how breast cancer risk is appraised. PMID- 15779053 TI - The relationship of maternal attributes, resources, and perceptions of postpartum experiences to depression. AB - The relationships of maternal attributes, resources, and perceptions of the postpartum experience to postpartum depression (PPD) were examined. One hundred and forty-three mothers completed measures of maternal attributes, resources, and perceptions of the postpartum experience, as well as depressed mood and clinical depression symptoms. Results from canonical correlation analysis revealed two patterns. Pattern 1: Women without history of depression, more emotional support from partners, higher parenting evaluation, lower centrality of the infant in the mother's thoughts and actions, and fewer life changes had lower depressed mood and fewer clinical depression symptoms than their counterparts. Pattern 2: Mothers without history of depression who were married, with higher financial stress, and lower parenting evaluation had higher depressed mood, but not more clinical depression symptoms. Maternal age, parity, time since delivery, income, and help from partner were unrelated to either depression indicator. These patterns suggest multiple paths to PPD, and the need for routine mental health assessment and exploration of women's perceptions of their postpartum experience. PMID- 15779054 TI - Test of the health promotion model as a causal model of commitment to a plan for exercise among Korean adults with chronic disease. AB - The purpose of this study was to test seven constructs (prior experience of exercise, perceived health status, exercise benefits, exercise barriers, exercise self-efficacy, social support for exercise, and options for exercise) from the health promotion model (HPM) as a causal model of commitment to a plan for exercise in a sample of 400 Korean adults with chronic disease. Using structural equation modeling (SEM), we found that all fit indices indicated a good fit. The final model accounted for 54% of the variance in commitment to a plan for exercise. Prior experience with exercise and exercise benefits were the factors most highly related. Health professionals can assess prior experience and emphasize personally relevant benefits of exercise in designing intervention programs to help Korean adults with chronic disease become more physically active. PMID- 15779055 TI - Work and retirement after cancer diagnosis. AB - A pilot telephone survey was administered to 253 cancer survivors, ages 35-75, 5 7 years after their diagnosis of breast, colorectal, lung, or prostate cancer. These survivors reported their demographic characteristics and, if they had retired, reasons for retirement and their impression of the retirement experience. Retirees were older, had less education and lower incomes, and fewer had health insurance through their current/former employer relative to employed survivors. Those who had retired, regardless of whether they retired before or after a cancer diagnosis, were similar in age, race/ethnicity, health insurance status, household income, and impressions of retirement. The results suggest that cancer may not negatively affect survivors' decision to retire or the quality of the retirement experience. PMID- 15779057 TI - Re: Predictors of patient satisfaction with inpatient hospital nursing care. PMID- 15779056 TI - Symptoms of unstable angina in patients with and without diabetes. AB - Patients with diabetes experience cardiac autonomic neuropathy that may affect the way they perceive the symptoms of unstable angina (UA). The purpose of this study was to examine symptom differences in patients with and without diabetes during an episode of UA. A convenience sample of 50 women and 50 men were recruited. Patients with diabetes were more likely to have a history of hypercholesterolemia (83% vs. 60%), prior history of heart disease (85% vs. 65%), and prior angiogram (85% vs. 67%). Patients with diabetes reported having less nausea (20% vs. 40%), less squeezing (25% vs. 48%) and less aching (25% vs. 45%) type pain, and more hyperventilation (27.5% vs. 11.7%). Other cardiac symptoms were similar between the groups. Further study of symptom presentation in patients with diabetes is warranted given their high levels of morbidity and mortality from cardiac disease. PMID- 15779058 TI - Health care and social service use among Chinese immigrant elders. AB - We explored patterns and reasons for health and social service use among Chinese immigrant elders. Interviews were conducted with 27 Chinese immigrant elders, 11 adult care giving children, and 12 health and social service providers. Content analysis of these data indicated that participants across groups agreed that Chinese elders under-utilize services because of problems related to language, transportation, cost, long waits for appointments, and because of cultural norms/values related to need for care, preference for self-over professional care, fear, and distrust of western biomedicine, and the obligation to refrain from using formal services. These problems are complicated by geographical dispersion and dialect differences in the local Chinese immigrant community. PMID- 15779059 TI - Positive and negative depression coping in low-income African American women. AB - The purpose of the study was to examine positive and negative depression coping (DC) in low-income African American women. Because low-income African American women have been shown to be vulnerable to depression symptom onset yet less accepting of treatment, DC in this population is of interest to researchers. Depression symptom severity, defense mechanisms, difficult life circumstances (DLC), and social support were examined as possible determinants of DC. In 244 mildly or moderately to severely depressed women, mature defense mechanisms predicted positive DC, and DLC predicted negative DC. Social support had no effect on positive or negative DC. Findings are discussed in terms of individual and community tailored rehabilitative psychotherapy to promote positive DC. PMID- 15779060 TI - Flow cytometric analysis of CD41-labeled platelets isolated by the rapid, one step OptiPrep method from human blood. AB - BACKGROUND: Although platelet-rich plasma is relatively easy to produce by centrifugation of whole blood, yields of platelets may be variable because many of them are trapped within the erythrocyte layer. Although they can be recovered by washing these cells, it is a general rule that the number of centrifugations should be kept to a minimum to avoid activation of platelets. This work describes the rapid, one-step OptiPrep method for the isolation of highly purified platelets from human blood (buffy coat). METHODS: To provide a functionally intact and uncontaminated platelet fraction, a density gradient centrifugation was performed by using a density barrier prepared from OptiPrep. CD41 antibody staining was performed to assess the purity of the obtained platelet population by means of a FACScan flow cytometer. Platelets were identified by a morphologic gate in which events were further studied for CD41 expression. Data were analyzed by CellQuest (Becton Dickinson). RESULTS: Platelet-specific CD41 antibody staining showed that the purity of the platelet population recovered from this density barrier method was greater than 90%. The platelets showed an excellent morphologic state. CONCLUSION: The rapid, one-step OptiPrep density gradient centrifugation is a reliable method for obtaining highly purified platelets from human blood that are ready for further pharmacologic investigations. PMID- 15779061 TI - On the merits of attention-control groups. PMID- 15779062 TI - Characterization of seven genotypes (A to E, G and H) of hepatitis B virus recovered from Japanese patients infected with human immunodeficiency virus type 1. AB - To investigate the prevalence of hepatitis B virus (HBV) genotypes and characteristics of HBV isolates among Japanese patients infected with human immunodeficiency virus type 1 (HIV), serum samples collected between September 1990 and March 2002 from 471 HIV-infected patients (age, 38.8 +/- 11.4 [mean +/- standard deviation] years; male, 90%) were tested for hepatitis B surface antigen (HBsAg) and HBV DNA. Positivity for HBsAg and HBV DNA was seen in 42 patients (8.9%), 41 of whom had contracted HIV infection through sexual activity and 1 had hemophilia. Genotypes of HBV were determined by comparative and phylogenetic analyses of the S gene sequence (396 nucleotides [nt]). The distribution of HBV genotypes among the 42 HBV-viremic patients was: A (50%), B (5%), C (24%), D (5%), E (2%), H (10%), A plus D (2%), A plus G (2%). The hemophilia patient had HBV genotype D. Genotypes E, G, and H which had not been reported in Japan, were found in one patient each who had traveled to Zambia, the US, and South America, respectively. Genotypes A and D, which are rare in Japan, were found in patients who had no history of traveling abroad. The entire genome of the HB-JI411 (genotype E [3,212 nt]), HB-JI444G (genotype G [3,248 nt]), and HB-JI260 (genotype H [3,218 nt]) isolates had the highest identity of 98.3%, 99.9%, and 98.5%, respectively, with reported HBV isolates of the same genotype. Most Japanese patients coinfected with HIV and HBV had HBV genotypes that are found rarely or had not been reported in Japan. PMID- 15779063 TI - DNA index, genome size, and electronic nuclear volume of vertebrates from the Miami Metro Zoo. AB - BACKGROUND: Flow cytometry is a rapid and reliable method for measuring nuclear DNA content and genome size. Fluorochrome binding characteristics, sample preparation and differences in DNA condensation, and availability of binding sites can cause variations in results obtained. METHODS: Blood samples from 82 vertebrate species were collected in 10% dimethyl sulfoxide and stained with propidium iodide/hypotonic citrate or 4,6-diamidino-2-phenylindole dihydrochloride for analysis of DNA content and electronic nuclear volume (ENV). Trout red blood cells (TRBCs), human peripheral blood lymphocytes, and human buccal cavity cells were used as internal standards. RESULTS: Mean fluorescence channel (MFC) values of TRBC and buccal cavity cells used as internal standards were stable at 15 to 120 min of propidium iodide staining. TRBCs mixed with other cells especially human peripheral blood cells showed an increase in MFC. ENV and MCF values were less variable in different species of birds than in reptiles or mammals. Genome size based on use of buccal cavity cells as the internal standard showed a high degree of correlation with previous reports. CONCLUSIONS: Proper selection and use of internal standards and sample preparation are essential for reliable determination of DNA content and genome size in vertebrates by flow cytometry. PMID- 15779064 TI - Studying the biological and technical sources of variation in telomere length of individual chromosomes. AB - BACKGROUND: Consistent average length differences between species and chromosome arm differences within species indicate that telomere length is genetically determined. This seems to contradict an observed large variation in lengths of the same human telomere between metaphases of the same individual. We examined the extent to which the variation in the telomeres of the human X and Y chromosomes is heritable, induced, or technical in origin. METHODS: Metaphase chromosomes were stained by fluorescence in situ hybridization with a telomere repeat-specific probe, and fluorescence intensities of the X and Y chromosomes were measured. If telomere length variation is predominantly genetically determined and a 50% probability of meiotic recombination between the pseudo autosomal regions of Yp and Xp in the father is taken into account, one expects an equal chance that the Yp telomere of a son is derived from his father's Xp or Yp telomere. This implies that the Yp/Yq telomere ratios in fathers and sons will be identical in the absence of paternal meiotic recombination and different when recombination occurs. RESULTS: Among five father-son pairs, four showed similar Yp/Yq ratios (P > 0.05), whereas one pair exhibited a large difference in the Yp/Yq ratio that was attributable to a significantly longer Xp than Yp telomere in the father and a presumptive meiotic exchange between X and Y during paternal meiosis. Further, the Xq telomere exhibited a generally shorter telomere length than the others. CONCLUSIONS: The high variation in telomere length appeared to be intracellular (between sister chromatids) and, hence, technical in nature. We found no measurable induced variation in the cells studied, implying that, if induced variation exists, it is small compared with the technical variation. PMID- 15779065 TI - Flow cytometric characterization of viable meiotic and postmeiotic cells by Hoechst 33342 in mouse spermatogenesis. AB - BACKGROUND: Spermatogenesis in adult is a complex stepwise process leading to terminally differentiated spermatozoa. The cellular heterogeneity of testis renders complex the studies on molecular aspects of this differentiation process. Analysis of the regulation of adult spermatogenesis would undoubtedly benefit from the development of techniques to characterize each germinal differentiation step. METHODS: Hoechst 33342 staining of mouse testicular cells allows characterization of an enriched population in germinal stem cell and spermatogonia, called side population. In this study, we examined the definition of the various germinal populations stained by Hoechst 33342, notably meiotic and postmeiotic cells. RESULTS: Preleptotene spermatocytes, spermatocyte I, spermatocyte II, and round and elongated spermatids were discriminated by Hoechst 33342 staining. In addition, we associated differentiation of spermatocyte I through leptotene to diplotene with changes in Hoechst 33342 red fluorescence pattern. CONCLUSIONS: Hoechst 33342 staining of viable germinal cells constitutes a valuable tool to study normal and impaired mouse adult spermatogenesis or to isolate viable cells from various differentiation stages for studies of molecular mechanisms regulating spermatogenesis. PMID- 15779066 TI - Maintaining your focus in focus groups: avoiding common mistakes. AB - Focus groups are used frequently in nursing research as a method for collecting qualitative data. However, due to inadequate planning or execution, the results can be of poor quality or even worthless. A well-defined purpose, carefully planned environment, and well trained personnel are discussed as the cornerstones to focus group success. This article sets the stage with a description of a well conducted focus group, then provides examples and a critique of less-than effective groups. Focus groups can be powerful and effective when their use is appropriate to the study purpose, the environment is non-judgmental and welcoming, and the personnel are skilled. PMID- 15779067 TI - Bioactivity and metabolism of trans-resveratrol orally administered to Wistar rats. AB - The purpose of this study was to investigate the implications of selected chemopreventive parameters and metabolic conversion of resveratrol in vivo. In two 8-week long feeding experiments with rats, a low-resveratrol diet containing 50 mg resveratrol per kg body weight (bw) and day and a high-resveratrol diet with 300 mg per kg bw and day were administered. For chemopreventive evaluation selected phase I and phase II enzymes of the biotransformation system, the total antioxidant activity, and the vitamin E status of the animals were determined. The level of resveratrol and its metabolites in the feces, urine, plasma, liver, and kidneys was identified and quantitated by high-performance liquid chromatography-diode array detection (HPLC-DAD) using synthesized resveratrol conjugate standards. Feeding of different dosages of resveratrol revealed no effect on the different chemopreventive parameters, except for the total antioxidant activity, which was elevated in plasma by 19% after feeding 50 mg resveratrol per kg bw and day. The formation of trans-resveratrol-3-sulfate, trans-resveratrol-4'-sulfate, trans-resveratrol-3,5-disulfate, trans-resveratrol 3,4'-disulfate, trans-resveratrol-3,4',5-trisulfate, trans-resveratrol-3-O-beta-D glucuronide, and resveratrol aglycone was detected by HPLC analysis, depending on the biological material. Total resveratrol recovery in urine and feces of rats fed on 50 mg resveratrol per kg bw and day was 15% and 13%, respectively. For rats fed the higher dosage of 300 mg resveratrol per kg bw and day recovery was 54% and 17%, respectively. This is the first study performed with synthesized standards of relevant resveratrol conjugates. The lack of effect on the chemopreventive parameters is probably due to the formation of various resveratrol conjugates reducing its bioavailability in the rat. PMID- 15779068 TI - Effects of resveratrol, piceatannol, tri-acetoxystilbene, and genistein on the inflammatory response of human peripheral blood leukocytes. AB - Inflammatory processes are involved in the etiology of diseases. We analyzed the effect of resveratrol, piceatannol, synthetic tri-acetoxystilbene (TAS), and genistein (Bonistein(TM)) on the production of inflammatory mediators including prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-alpha), interleukins, and chemokines, which participate in the progression of inflammation. In order to induce inflammatory responses, human peripheral blood mononuclear and/or polymorphonuclear leukocytes were stimulated with lipopolysaccharide (LPS) and interferon-gamma (IFN(gamma)) and the production of PGE2, interleukin-8 (IL-8), and TNF-alpha was determined. In response to the stimuli, genes were substantially activated within < 2 h (e. g., TNF-alpha, IL-1alpha), or at a later stage, (e. g., COX-2, IL-6, IL-8). Unlike genistein, resveratrol and related compounds dose-dependently reduced PGE2 production. Genistein, piceatannol, and TAS diminished secretion of TNF-alpha, and IL-8. TAS reduced mRNA levels of COX 2, TNF-alpha, IL-8, IL-6, and IL-1alpha, while resveratrol impaired early expression of IL-8 and TNF-alpha. Piceatannol out-performed resveratrol, yet without matching TAS. Genistein downregulated TNF-alpha and IL-8 expression. These substances altered the LPS/IFNgamma-induced gene expression in mononuclear cells rather than in polymorphonuclear leukocytes. Immunoblot analyses corroborated the distinct activity pattern of resveratrol and genistein. In conclusion, resveratrol and their derivatives attenuated the inflammatory response of PBLs at several levels, whereas genistein acts on cytokines and pro inflammatory interleukins. PMID- 15779069 TI - Prevalence of vertebral alterations and the effects of calcium and vitamin D supplementation on calcium metabolism and bone mineral density after gastrectomy. AB - BACKGROUND: Bone disease is common after gastrectomy, resulting in decreased bone mass and an increased risk of fracture. No proven therapy is currently available. METHODS: Serum markers of calcium metabolism in 98 patients after partial or total gastrectomy were compared with those in 30 age- and sex-matched healthy controls. Patients with disorders of calcium metabolism were investigated by conventional radiography and single-energy computed tomography of the spine. Forty patients participated in a 1-year follow-up study to investigate the effects of vitamin D and calcium supplementation on calcium metabolism and bone mineral density. RESULTS: Altered serum markers of calcium and phosphate metabolism were observed in 77 (79 per cent) of 98 patients. Sixty (79 per cent) of these had vertebral alterations. Vertebral fractures were detected in 22 patients, grade I vertebral deformities in 50 patients, grade II deformities in 22 patients and osteopenia (Z-score less than - 1) in 30 patients. Calcium and vitamin D supplementation resulted in an increase in 25-hydroxy-vitamin D (P < 0.001), 1,25-dihydroxy-vitamin D (P = 0.048) and osteocalcin (P = 0.045), whereas levels of parathyroid hormone were decreased (P = 0.007). Bone mineral density did not change over time. CONCLUSION: Disturbances of calcium and bone metabolism are common after gastrectomy. Calcium and vitamin D supplementation normalized levels of markers of calcium metabolism and might have prevented age-related bone mass loss, although it did not increase bone mineral density after 1 year. PMID- 15779070 TI - Metabolism and bioavailability of trans-resveratrol. AB - Resveratrol (3,4',5-trihydroxy-trans-stilbene) is a polyphenolic compound accounting to the stilbene class. Most stilbenes in plants act as antifungal phytoalexins, compounds that are usually synthesized only in response to infection or injury. Resveratrol has been detected in trees, in a few flowering plants, in peanuts, and in grapevines. The major dietary sources of resveratrol include grapes, wine, peanuts, and peanut products. Numerous in vitro studies describe different biological effects of resveratrol. The major impacts are the antioxidative, anti-inflammatory, and estrogenic effects as well as anticancer and chemopreventive activities. In order to reveal information on absorption, metabolism, and the consequent bioavailability of resveratrol, different research approaches were performed, including in vitro, ex vivo, and in vivo models, all of which are considered in this review. Summarizing the data, resveratrol is absorbed and metabolized. Around 75% of this polyphenol are excreted via feces and urine. The oral bioavailability of resveratrol is almost zero due to rapid and extensive metabolism and the consequent formation of various metabolites as resveratrol glucuronides and resveratrol sulfates. The potential biologic activity of resveratrol conjugates should be considered in future investigations. PMID- 15779071 TI - Co-morbidity leads to altered treatment and worse survival of elderly patients with colorectal cancer. AB - BACKGROUND: The aim of this study was to evaluate the effects of co-morbidity on the treatment and prognosis of elderly patients with colorectal cancer. METHODS: The independent influence of age and co-morbidity on treatment and survival was analysed for 6931 patients with colorectal cancer aged 50 years or more diagnosed between 1995 and 2001 in the southern part of the Netherlands. RESULTS: Co morbidity had no influence on resection rate. The use of adjuvant chemotherapy in patients with stage III colonic cancer was influenced by co-morbidity, especially a previous malignancy (odds ratio (OR) 0.2 (95 per cent confidence interval (c.i.) 0.1 to 0.6); P = 0.002) or chronic obstructive pulmonary disease (COPD) (OR 0.3 (95 per cent c.i. 0.1 to 0.9); P = 0.043). Co-morbidity also influenced use of adjuvant radiotherapy in patients with rectal cancer, especially the presence of hypertension in combination with diabetes (OR 0.5 (95 per cent c.i. 0.2 to 0.9); P = 0.031). Co-morbidity influenced survival (hazard ratio up to 1.6), when adjusted for age, sex, tumour stage and treatment. The greatest influence on survival of patients with colonic cancer was previous malignancy, cardiovascular disease and COPD, and that of patients with rectal cancer was COPD, hypertension, and hypertension in combination with diabetes. CONCLUSION: Elderly patients with co-morbidity were treated less aggressively and had a worse survival than those with no concomitant disease. PMID- 15779072 TI - EXternal Pelvic REctal SuSpension (Express procedure) for rectal intussusception, with and without rectocele repair. AB - BACKGROUND: The results of conventional treatment for rectal intussusception and rectocele are unpredictable. The aim was to develop a less invasive surgical approach and to evaluate outcome in selected patients. METHODS: Seventeen patients (13 women; median age 47 (range 20-67) years) with rectal evacuatory dysfunction and rectal intussusception, 13 of whom had a rectocele, were selected. The intussusception was corrected by external pelvic suspension of the rectum, using collagen strips attached to the rectal wall and pubis. The rectocele was repaired with a collagen patch. Patients were assessed before and 6 months after surgery by symptom and quality of life questionnaires, anorectal physiological investigation and proctography, and were followed up for a median of 12 months. RESULTS: Sepsis requiring exploration occurred in two patients but there was no extrusion or need to remove the collagen. Of the 15 patients assessed after surgery, total symptom scores were significantly decreased (P < 0.001) and quality of life scores improved (P < 0.001). Proctographically, the degree of intussusception was improved in ten patients; six patients had normal postoperative proctograms. The rectocele was reduced in size in all patients, and was not demonstrable in eight. CONCLUSION: An effective procedure for rectal intussusception and rectocele has been developed in a selected group of patients with marked evacuatory symptoms. PMID- 15779073 TI - Variability in food-processing behavior among white-faced capuchins (Cebus capucinus) in Santa Rosa National Park, Costa Rica. AB - Observed patterns of variability in the food-processing behavior of white-faced capuchins (Cebus capucinus) across populations may reflect foraging traditions. However, there has been relatively little attention given to intrapopulation variability in food processing among groups and age/sex classes, making recent cross-population comparisons difficult to interpret. In this paper, we provide data on patterns of object use in foraging that we observed at Santa Rosa National Park, Costa Rica, for comparison with published data from a neighboring research site, Palo Verde National Park. We also describe the techniques used to process two food items consumed by Cebus capucinus at Santa Rosa, and discuss the factors that may underlie observed variability. We conducted a 6-month study on two groups of capuchins in 2001, and collected data on general activity and feeding patterns, rates and forms of object use, and distinctive processing techniques employed for two specific foods (Sloanea terniflora and Luehea candida). Rates of object-use behavior at Santa Rosa were considerably higher than those reported for Palo Verde and showed significant variation between groups and age/sex classes, as did patterns of Sloanea and Luehea processing. Observed differences in feeding rates between groups may reflect food availability or relative profitability, whereas variation between age/sex classes seems to reflect differences in the physical capabilities, foraging strategies, and the relative experience of mature and immature animals. Further research is needed to identify how a social context may influence the acquisition of food processing techniques in juveniles and the development of foraging traditions in social groups. PMID- 15779074 TI - Doppler assessment after right hepatectomy confirms the need to fix the remnant left liver in the anatomical position. AB - BACKGROUND: The remnant left liver after right hepatectomy tends to rotate spontaneously into the right subphrenic space. This rotation might induce venous outflow impairment. The aim of this study was to assess immediate venous outflow in the left hepatic vein by intraoperative Doppler ultrasound (US) according to the position of the remnant liver. METHODS: From August 2003 to February 2004, assessment of left hepatic venous outflow was systematically performed in 44 consecutive right hepatic resections by Doppler US in spontaneous and anatomical positions. The anatomical position was defined as the position in which the falciform ligament was in its strict median position. RESULTS: The placement of the left liver from the spontaneous position to the anatomical position resulted in a significant increase in left hepatic venous outflow (20.1 +/- 5.7 versus 8.5 +/- 4.4 cm/s; P < 0.0001). In the spontaneous position, the decrease in left hepatic venous outflow persisted even without division of the left triangular ligament (10.2 +/- 5.4 versus 21.7 +/- 5.3 cm/s in the anatomical position) or removal of the middle hepatic vein (8.4 +/- 3.4 versus 21.3 +/- 5.8 cm/s). CONCLUSION: : Results of this study strongly suggest that after right hepatectomy the remnant left liver should always be fixed in the anatomical position. PMID- 15779075 TI - Increased calcitonin level in thyroid nodules without medullary carcinoma. AB - BACKGROUND: Basal calcitonin measurement is routinely performed in patients with a thyroid nodule to detect medullary carcinoma. However, increased calcitonin does not always correlate with medullary carcinoma. The aim of this study was to analyse increased calcitonin levels in patients without medullary carcinoma and to find out whether absence of this carcinoma can be predicted with certainty. METHODS: From 1992 to 2003, 5018 patients with thyroid nodules underwent thyroid surgery. A retrospective analysis of preoperative increased calcitonin levels in 67 of these patients was performed. RESULTS: Pathology revealed medullary carcinoma in 16 patients (group I), micromedullary carcinoma in 13 (group II) and no medullary carcinoma in 38 (group III). In group III, 30 patients had C-cell hyperplasia. The mean basal calcitonin level was 6250 pg/ml in group I (39-62 500), 109.6 pg/ml in group II (10-728) and 25.5 pg/ml in group III (10.5-145). The mean pentagastrin-stimulated calcitonin level was 1074.1 pg/ml in group II (26-5700) and 67.6 pg/ml in group III (10-205). CONCLUSION: There is an overlap of thyroid C-cell pathology for medullary carcinoma, micromedullary carcinoma and C-cell hyperplasia that occurs when basal calcitonin is between 10 and 145 pg/ml and pentagastrin-stimulated calcitonin between 10 and 205 pg/ml. In these patients, since medullary carcinoma cannot be completely excluded, total thyroidectomy should be recommended. PMID- 15779076 TI - Heritability of age at menarche in girls from the Fels Longitudinal Study. AB - Menarche is the hallmark maturational event of female childhood. Many studies indicated a significant genetic contribution to the timing of the onset of menstruation, but most of these studies were limited by the use of retrospective data and by the use of data from only certain types of relatives (i.e., mothers and daughters, sisters, or twin sisters). The primary goal of this study was to use a modern maximum likelihood quantitative genetic method to estimate the heritability (h(2)) of age at menarche, using familial data collected over the course of the 74-year-old Fels Longitudinal Study. The secondary goal was to review earlier studies of the heritability of age at menarche. The study of the heritability of age at menarche presented here is unique for two reasons. First, because of the Fels Longitudinal Study's serial design, age-at-menarche data were collected prospectively from most participants. Second, because the Fels Longitudinal Study is a family study that has been conducted for decades, age-at menarche data are available from many types of female relatives spanning multiple households and generations. The best-fitting and most parsimonious quantitative genetic model included provision for a secular decrease in age at menarche, and estimated the h(2) of age at menarche to be 0.49+/- 0.13 (95% confidence interval of h(2),=0.24-0.73). The results of this study are in general agreement with the findings of most previous studies of genetic influences on age at menarche, and suggest that it is reasonable to consider it well-established that approximately half the phenotypic variation among girls from developed nations in the timing of menarche is due to genetic factors. PMID- 15779077 TI - In search of the vertebrate phylotypic stage: a molecular examination of the developmental hourglass model and von Baer's third law. AB - In 1828, Karl von Baer proposed a set of four evolutionary "laws" pertaining to embryological development. According to von Baer's third law, young embryos from different species are relatively undifferentiated and resemble one another but as development proceeds, distinguishing features of the species begin to appear and embryos of different species progressively diverge from one another. An expansion of this law, called "the hourglass model," has been proposed independently by Denis Duboule and Rudolf Raff in the 1990s. According to the hourglass model, ontogeny is characterized by a starting point at which different taxa differ markedly from one another, followed by a stage of reduced intertaxonomic variability (the phylotypic stage), and ending in a von-Baer-like progressive divergence among the taxa. A possible "translation" of the hourglass model into molecular terminology would suggest that orthologs expressed in stages described by the tapered part of the hourglass should resemble one another more than orthologs expressed in the expansive parts that precede or succeed the phylotypic stage. We tested this hypothesis using 1,585 mouse genes expressed during 26 embryonic stages, and their human orthologs. Evolutionary divergence was estimated at different embryonic stages by calculating pairwise distances between corresponding orthologous proteins from mouse and human. Two independent datasets were used. One dataset contained genes that are expressed solely in a single developmental stage; the second was made of genes expressed at different developmental stages. In the second dataset the genes were classified according to their earliest stage of expression. We fitted second order polynomials to the two datasets. The two polynomials displayed minima as expected from the hourglass model. The molecular results suggest, albeit weakly, that a phylotypic stage (or period) indeed exists. Its temporal location, sometimes between the first-somites stage and the formation of the posterior neuropore, was in approximate agreement with the morphologically defined phylotypic stage. The molecular evidence for the later parts of the hourglass model, i.e., for von Baer's third law, was stronger than that for the earlier parts. PMID- 15779078 TI - Long-term outcome after laparoscopic fenestration of symptomatic simple cysts of the liver. PMID- 15779079 TI - Role of laparoscopic cholecystectomy in the early management of acute gallbladder disease. AB - BACKGROUND: This study evaluated the role of laparoscopic surgery in the early management of acute gallbladder disease in a single large UK teaching hospital. METHODS: Details of all emergency admissions for acute gallbladder disease from January 2000 to December 2001 were identified and additional information from the hospital records was reviewed retrospectively. RESULTS: Three hundred and eighty five patients with gallstone disease (243 acute biliary pain, 142 acute cholecystitis) and 15 with acalculous disease were identified. The conversion rate was higher during early laparoscopic surgery for acute calculous cholecystitis than in operations for acute biliary pain (19 versus 4 per cent; P = 0.002). In patients with acute calculous cholecystitis the conversion rate was significantly lower in operations within 48 h of admission (one of 26) than when surgery was delayed beyond 48 h (14 of 52) or subsequently carried out electively (seven of 21) (P = 0.014). Elective surgery for previous acute cholecystitis was associated with a higher conversion rate (seven of 21 patients) than elective surgery for biliary pain (three of 65) (P = 0.002). CONCLUSION: Laparoscopic cholecystectomy for acute calculous cholecystitis should be performed, where possible, within the first 48 h of admission. PMID- 15779080 TI - Microsurgery and the reconstructive ladder. PMID- 15779081 TI - Circular dichroism studies of carbopeptoid-cyclodextrins. AB - A series of sugar amino acids, based on open chain sugars, have been oligomerised and cyclised. The resulting cyclic carbopeptoids have been examined for desirable properties such as host-guest chemistry (as in cyclodextrins) or self-assembling properties (e.g., peptide nanotubes). Initial studies of these systems, by circular dichroism and X-ray crystallography, have given valuable insight into their stability and properties. One of the four cyclic species studied was found to interact with ion/molecular probes. PMID- 15779082 TI - Development of the commercial process for Zoloft/sertraline. AB - Sertraline represented Pfizer's first product with stereocenters not derived from natural sources, as was the case with azithromycin and beta-lactam products. Discovery chemists employed chromatography and classical resolution to secure the active agent's two stereocenters. Investigations to identify a commercial route comprised a number of synthetic approaches, including enantioselective ketone reduction, SN2 displacement of an activated alcohol to transfer chirality, and continuous chromatography. The challenges of adopting new technology in the pharmaceutical industry are discussed in the context of sertraline's development. The successful outcome employed continuous chromatography, and the potential for this technology to shift the development paradigm for any development candidate or commercial product is described. PMID- 15779083 TI - Chymotrypsin-like proteases contribute to human monocytic THP-1 cell as well as human microglial neurotoxicity. AB - Activated microglia have been observed in various neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis, and multiple sclerosis. They may exacerbate neuronal damage by secreting various toxic molecules. The list of candidate toxins includes proteases. Since it is currently not known which, if any, proteases are involved in human microglia neurotoxicity, we studied the effects of a panel of protease inhibitors on the toxicity of cell-free supernatants of stimulated human microglia and THP-1 monocytic cells to human SH-SY5Y cells. Five structurally distinct inhibitors that are known to inhibit chymotrypsin-like proteases were partially protective. They included chymostatin, AEBSF (Pefabloc SC), alpha1 antichymotrypsin, bromoenol lactone, and 3,4-dichloroisocoumarin. The data suggest that certain protease inhibitors could inhibit microglial-mediated toxicity. They might represent a novel class of drugs with benefit in diseases where overactivity of microglia contributes to the pathogenesis. PMID- 15779084 TI - Very low intensity alternating current decreases cell proliferation. AB - Electric fields impact cellular functions by activation of ion channels or by interfering with cell membrane integrity. Ion channels can regulate cell cycle and play a role in tumorigenesis. While the cell cycle may be directly altered by ion fluxes, exposure to direct electric current of sufficient intensity may decrease tumor burden by generating chemical products, including cytotoxic molecules or heat. We report that in the absence of thermal influences, low frequency, low-intensity, alternating current (AC) directly affects cell proliferation without a significant deleterious contribution to cell survival. These effects were observed in normal human cells and in brain and prostate neoplasms, but not in lung cancer. The effects of AC stimulation required a permissive role for GIRK2 (or K(IR)3.2) potassium channels and were mimicked by raising extracellular potassium concentrations. Cell death could be achieved at higher AC frequencies (>75 Hz) or intensities (>8.5 microA); at lower frequencies/intensities, AC stimulation did not cause apoptotic cellular changes. Our findings implicate a role for transmembrane potassium fluxes via inward rectifier channels in the regulation of cell cycle. Brain stimulators currently used for the treatment of neurological disorders may thus also be used for the treatment of brain (or other) tumors. PMID- 15779086 TI - Activation of catechol-O-methyltransferase in astrocytes stimulates homocysteine synthesis and export to neurons. AB - Elevation of the total homocysteine (tHcy) concentration in plasma has been implicated in neurodegeneration in patients with stroke, dementia, Alzheimer disease, and Parkinson disease. Because the mechanisms controlling brain tHcy are unknown, the present study investigated its synthesis and transport in primary rat brain cell cultures. We found that the catechol-O-methyltransferase (COMT) substrate 3,4-dihydroxybenzoic acid (DHB) increased export of tHcy in astrocytes, but not in neurons. The export mechanism was selective for tHcy over cyst(e)ine, total glutathione (tGSH) or cysteinylglycine (Cys-Gly). tHcy export from astrocytes was also induced by the COMT substrates levodopa (L-DOPA), dopamine and quercetin, and it was blocked by the COMT inhibitors tropolone and entacapone. This export was associated with increased synthesis of tHcy because both intracellular and extracellular tHcy concentrations rose during COMT activation. Incubation in cyst(e)ine-deficient medium inhibited the tHcy export response to COMT activation. Exogenous tHcy (100 muM) was accumulated into neurons, but not into astrocytes. We conclude that activation of COMT causes sustained synthesis of Hcy in astrocytes and transport of this amino acid to neurons. PMID- 15779085 TI - Mechanical strain injury increases intracellular sodium and reverses Na+/Ca2+ exchange in cortical astrocytes. AB - Traditionally, astrocytes have been considered less susceptible to injury than neurons. Yet, we have recently shown that astrocyte death precedes neuronal death in a rat model of traumatic brain injury (TBI) (Zhao et al.: Glia 44:140-152, 2003). A main mechanism hypothesized to contribute to cellular injury and death after TBI is elevated intracellular calcium ([Ca2+]i). Since calcium regulation is also influenced by regulation of intracellular sodium ([Na+]i), we used an in vitro model of strain-induced traumatic injury and live-cell fluorescent digital imaging to investigate alterations in [Na+]i in cortical astrocytes after injury. Changes in [Na+]i, or [Ca2+]i were monitored after mechanical injury or L glutamate exposure by ratiometric imaging of sodium-binding benzofuran isophthalate (SBFI-AM), or Fura-2-AM, respectively. Mechanical strain injury or exogenous glutamate application produced increases in [Na+]i that were dependent on the severity of injury or concentration. Injury-induced increases in [Na+]i were significantly reduced, but not completely eliminated, by inhibition of glutamate uptake by DL-threo-beta-benzyloxyaspartate (TBOA). Blockade of sodium dependent calcium influx through the sodium-calcium exchanger with 2-[2-[4-(4 Nitrobenzyloxy)phenyl]ethyl]isothiourea mesylate (KB-R7943) reduced [Ca2+]i after injury. KB-R7943 also reduced astrocyte death after injury. These findings suggest that in astrocytes subjected to mechanical injury or glutamate excitotoxicity, increases in intracellular Na+ may be a critical component in the injury cascade and a therapeutic target for reduction of lasting deficits after traumatic brain injury. PMID- 15779087 TI - Involvement of phospholipase A2 and lipoxygenase in lipopolysaccharide-induced inducible nitric oxide synthase expression in glial cells. AB - The present study underlines the importance of phospholipase A2 (PLA2)- and lipoxygenase (LO)-mediated signaling processes in the regulation of inducible nitric oxide synthase (iNOS) gene expression. In glial cells, lipopolysaccharide (LPS) induced the activities of PLA2 (calcium-independent PLA2; iPLA2 and cytosolic PLA2; cPLA2) as well as gene expression of iNOS. The inhibition of cPLA2 by methyl arachidonyl fluorophosphates (MAFP) or antisense oligomer against cPLA2 and inhibition of iPLA2 by bromoenol lactone reduced the LPS-induced iNOS gene expression and NFkappaB activation. In addition, the inhibition of LO by nordihydroguaiaretic acid (NDGA; general LO inhibitor) or MK886 (5-LO inhibitor), but not baicalein (12-LO inhibitor), completely abrogated the LPS-induced iNOS expression. Because NDGA could abrogate the LPS-induced activation of NFkappaB, while MK886 had no effect on it, LO-mediated inhibition of iNOS gene induction by LPS may involve an NFkappaB-dependent or -independent (by 5-LO) pathway. In contrast to LO, however, the cyclooxygenase (COX) may not be involved in the regulation of LPS-mediated induction of iNOS gene because COX inhibition by indomethacin (general COX inhibitor), SC560 (COX-1 inhibitor), and NS398 (COX-2 inhibitor) affected neither the LPS-induced iNOS expression nor activation of NFkappaB. These results indicate a role for cPLA2 and iPLA2 in LPS-mediated iNOS gene induction in glial cells and the involvement of LO in these reactions. PMID- 15779088 TI - Expression of a dominant negative IFN-gammareceptor on mouse oligodendrocytes. AB - The interferon-gamma (IFN-gamma) receptor is expressed by all nucleated cells, and binding of its cognate ligand, IFN-gamma, induces a wide variety of biological functions. Transgenic mice expressing a dominant negative IFN-gamma receptor 1 (IFN-gammaR1DeltaIC) on oligodendrocytes under control of the myelin proteolipid protein promoter are described. The mRNA encoding the transgene was only detected in the nervous system and protein expression was confirmed by immunohistochemistry. Transgenic receptor expression does not alter myelination and the mice exhibited no clinically apparent phenotype. Consistent with the restricted nervous system expression of the transgene, no alterations in peripheral immune responses were detected. Flow cytometric analysis demonstrated constitutive expression of both the IFN-gammaR1DeltaIC transgene and the endogenous IFN-gamma receptor 2 at high levels on oligodendrocytes derived from the transgenic mice. These oligodendrocytes also exhibited decreased STAT1 phosphorylation in response to IFN-gamma, confirming dominant negative transgene function. Transgenic mice in which oligodendrocytes have a diminished ability to respond to IFN-gamma showed delayed virus clearance from oligodendroglia compared with wild-type mice. This model will allow evaluation of oligodendrocyte responses to this critical cytokine during CNS inflammation. PMID- 15779089 TI - Downregulation of GFAP, TSP-1, and p53 in human glioblastoma cell line, U373MG, by IE1 protein from human cytomegalovirus. AB - Human cytomegalovirus (HCMV) is a member of the beta-herpesvirus family, which has tropism for glial cells. It was recently reported that HCMV might play important roles in the pathogenesis of malignant glioma. In this study, we investigated the effects of the HCMV IE1 protein on the gene expression profile in the human glioblastoma cell line, U373MG by employing cDNA microarray technology. Using DNA chips containing approximately 1,000 human cDNAs, RNA samples from U373MG cells stably expressing IE1 were compared with those from the control cells lacking IE1 cDNA. Fluorescence intensities of 13 genes were significantly decreased in IE1-expressing cells, while one gene was found to be upregulated. Among these 14 genes, we chose to work further on glial fibrillary acidic protein (GFAP), thrombospondin-1 (TSP-1), and p53, because of their previously known involvement in tumorigenesis. The mRNA levels of all these genes were found to be decreased in IE1-expressing glioblastoma cells by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) as well as Northern blot analysis. The decreased expression of these genes was also observed at protein levels as measured by immunocytochemistry or fluorescence-activated cell sorting (FACS) analysis. Our data strongly suggested that HCMV IE1 could modulate the expression of cellular genes that might play important roles in the pathogenesis of glial tumors. PMID- 15779090 TI - Leukemia inhibitory factor promotes oligodendrocyte survival after spinal cord injury. AB - Injury to the mammalian spinal cord is accompanied by a delayed, secondary wave of oligodendrocyte apoptosis that arises several days after the initial injury. A strong candidate to support oligodendrocyte survival after spinal cord injury is the pleiotropic cytokine, leukemia inhibitory factor (LIF). In vitro, LIF potentiates the differentiation and survival of oligodendrocyte precursors. LIF can also prevent oligodendrocyte apoptosis in response to either growth factor removal or cytotoxic challenge. More recently, in vivo studies have demonstrated that LIF is effective in preventing oligodendrocyte death in a mouse model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE). We therefore asked whether systemic delivery of LIF could ameliorate oligodendrocyte death in a mouse model of spinal cord injury. We have found that daily administration of LIF (25 microg/kg/day) promotes oligodendrocyte survival after spinal cord injury. Interestingly however, this effect does not appear to be mediated by a direct action of LIF on the oligodendrocyte but rather via an ancillary cell type, which results in augmented expression of another trophic factor capable of supporting oligodendrocyte survival, insulin-like growth factor 1 (IGF-1). PMID- 15779091 TI - Using baseline data to design a group randomized trial. AB - Group randomized trials (GRT) are often designed with relatively little preliminary data available to estimate key parameters. In this paper, however, the opposite situation is considered-very good baseline data are available on the primary outcome of interest. These data can then be used to inform key design and analysis decisions such as (i) should the trial be designed as an unmatched or pair-matched study, or stratified in some other fashion; (ii) is analysis of "change from baseline" preferable to using end-of-study data alone; and (iii) what power might be expected by pursuing these various strategies. The results are applied to a GRT for sexually transmitted diseases prevention in Peru. PMID- 15779092 TI - Covalent hemin-DNA adducts for generating a novel class of artificial heme enzymes. PMID- 15779093 TI - Intramolecular hydroarylation of alkynes catalyzed by platinum or gold: mechanism and endo selectivity. AB - The cyclization of differently substituted aryl alkynes with PtII or AuI catalysts proceeds by endo-dig pathways. When AgI was used to generate reactive cationic AuI catalysts, 2H-chromenes dimerize to form cyclobutane derivatives by a AgI-catalyzed process. A DFT study on the cyclization mechanism shows a kinetic and thermodynamic preference for 6-endo-dig versus 5-exo-dig cyclizations in PtII catalyzed processes. Calculations indicate that although Friedel-Crafts and the cyclopropanation processes via metal cyclopropyl carbenes show very similar activation energies, platinum cyclopropyl carbenes are the stationary points with the lowest energy. PMID- 15779094 TI - Tuning the decay time of lanthanide-based near infrared luminescence from micro- to milliseconds through d-->f energy transfer in discrete heterobimetallic complexes. AB - Inert and optically active pseudo-octahedral Cr(III)N6 and Ru(II)N6 chromophores have been incorporated by self-assembly into heterobimetallic triple-stranded helicates HHH-[CrLnL3]6+ and HHH-[RuLnL3]5+. The crystal structures of [CrLnL(3)](CF(3)SO(3))(6) (Ln=Nd, Eu, Yb, Lu) and [RuLnL3](CF3SO3)5 (Ln=Eu, Lu) demonstrate that the helical structure can accommodate metal ions of different sizes, without sizeable change in the intermetallic MLn distances. These systems are ideally suited for unravelling the molecular factors affecting the intermetallic nd-->4f communication. Visible irradiation of the Cr(III)N6 and Ru(II)N6 chromophores in HHH-[MLnL3]5/6+ (Ln=Nd, Yb, Er; M=Cr, Ru) eventually produces lanthanide-based near infrared (NIR) emission, after directional energy migration within the complexes. Depending on the kinetic regime associated with each specific d-f pair, the NIR luminescence decay times can be tuned from micro- to milliseconds. The origin of this effect, together with its rational control for programming optical functions in discrete heterobimetallic entities, are discussed. PMID- 15779095 TI - Gas-phase catalysis by atomic and cluster metal ions: the ultimate single-site catalysts. AB - Gas-phase experiments with state-of-the-art techniques of mass spectrometry provide detailed insights into numerous elementary processes. The focus of this Review is on elementary reactions of ions that achieve complete catalytic cycles under thermal conditions. The examples chosen cover aspects of catalysis pertinent to areas as diverse as atmospheric chemistry and surface chemistry. We describe how transfer of oxygen atoms, bond activation, and coupling of fragments can be mediated by atomic or cluster metal ions. In some cases truly unexpected analogies of the idealized gas-phase ion catalysis can be drawn with related chemical transformations in solution or the solid state, and so improve our understanding of the intrinsic operation of a practical catalyst at a strictly molecular level. PMID- 15779096 TI - Direct synthesis of an oligonucleotide-poly(phenylene ethynylene) conjugate with a precise one-to-one molecular ratio. PMID- 15779097 TI - Why Jesse likes Jennifer. PMID- 15779098 TI - By the way, doctor. My sister passed away from a brain aneurysm several months ago. She was just 33. My doctor says I have nothing to worry about because it's not hereditary. Still, I'm worried. Do I and other close relatives need to be checked? PMID- 15779099 TI - HIV and gender. Health research supports call for increased HIV testing. PMID- 15779100 TI - Claim of HIV dementia might free convicted murderer. PMID- 15779101 TI - New resistant strain of HIV worries health officials. PMID- 15779102 TI - HIV-discrimination claim fails for man with two jobs. PMID- 15779103 TI - Prisons. DOC officials cleared of delayed HIV testing for injured inmate. PMID- 15779104 TI - Prisons. BOP to pay $1.1 million award for false AIDS-suicide claim. PMID- 15779105 TI - Abstracts from the 2005 Tandem Bone and Marrow Transplantation Meetings. February 10-14, 2005, Keystone, Colorado, USA. PMID- 15779106 TI - Proceedings of the 30th Annual Scientific Meeting of the Society of Interventional Radiology. March 31-April 5, 2205, New Orleans, Louisiana, USA. PMID- 15779107 TI - Use of hormonal replacement therapy after treatment of breast cancer. Number 142, May 2004. AB - OBJECTIVE: To review the use of hormonal replacement therapy (HRT) after treatment of breast cancer. OPTIONS: The effect and role of estrogens on breast cancer. OUTCOME: Improved health and quality of life for women with breast cancer. VALUES: References were collected through MEDLINE searches up to 2002. EVIDENCE: The level of evidence and quality of recommendations have been determined using the criteria described by the Canadian Task Force on the Periodic Health Examination. BENEFITS, HARMS, AND COSTS: Utilization of the information to make proper risk-benefit assessment of HRT use in women with breast cancer. RECOMMENDATIONS: 1. HRT after treatment of breast cancer has not been demonstrated to have an adverse impact on recurrence and mortality. (II-2B). 2. HRT is an option in postmenopausal women with previously treated breast cancer. (II-2B). 3. Prospective, randomized clinical trial results are needed. (III-A). VALIDATION: Recommendations were viewed and revised by the Breast Diseases Committee of the Society of Obstetricians and Gynaecologists of Canada (SOGC) and approved by the Executive and Council of the SOGC. SPONSOR: The Society of Obstetricians and Gynaecologists of Canada. PMID- 15779108 TI - William Trager. PMID- 15779109 TI - SOGC clinical practice guidelines. The detection and management of vaginal atrophy. Number 145, May 2004. AB - OBJECTIVE: To support the practitioner in the diagnosis of vaginal atrophy and in the management of the related symptoms. OPTIONS: The modalities of evaluation range from basic pelvic examination, examination of the vulva, and laboratory tests. OUTCOMES: A comprehensive approach to the detection of vaginal atrophy and a discussion of available therapeutic and nontherapeutic options. EVIDENCE: Published opinions of experts, supplemented by evidence from clinical trials, where appropriate. VALUES: The quality of the Force on the Periodic Health Examination. BENEFITS, HARMS AND COSTS: Diagnosis of vaginal atrophy is often a challenge because women are unwilling to report symptoms, which have the potential to significantly decrease their quality of life. Increased clinical suspicion is the first step in the diagnosis of vaginal atrophy, which will prompt the initiation of safe therapies with proven efficacy. RECOMMENDATIONS: (1) Health-care providers should routinely assess postmenopausal women for the symptoms and signs of vaginal atrophy, a common condition that exerts significant negative effects on quality of life. (III-C) (2) Regular sexual activity should be encouraged to maintain vaginal health. (II-2B) (3) Women experiencing recurrent urinary tract infections should be instructed that consumption of pure cranberry-lingonberry juice, rather than cranberry drink, will decrease their risk of urinary infections. (I-A) (4) Vaginal moisturizers applied on a regular basis have an efficacy equivalent to local hormone replacement for the treatment of local urogenital symptoms such as vaginal itching, irritation, and dyspareunia, and should be offered to women wishing to avoid the use of hormone replacement therapy. (I-A) (5) Women experiencing vaginal atrophy can be offered any of the following effective vaginal estrogen replacement therapies: conjugated equine estrogen (CEE) cream (I-A), a sustained-release intravaginal estradiol ring (I-A), or a low-dose estradiol tablet. (I-A) (6) Although systematic absorption of estrogen can occur with local preparations, there is insufficient data to recommend annual endometrial surveillance in asymptomatic women using local estrogens. (III-C) (7) For menopausal women experiencing recurrent urinary tract infections and who have no contraindication to local hormone replacement, vaginal estrogen therapy should be offered. (I-A) VALIDATION: These guidelines have been reviewed by the joint committee of Clinical Practice Gynaecology and Urogynaecology and approved by the Execute and Council of the Society of Obstetricians and Gynaecologists of Canada. SPONSOR: The Society of Obstetricians and Gynaecologists of Canada. PMID- 15779110 TI - Guidelines for operative vaginal birth. Number 148, May 2004. AB - OBJECTIVE: To provide guidelines for operative vaginal birth in the management of the second state of labour. OPTIONS: Nonoperative techniques, episiotomy, and Caesarean section are compared to operative vaginal birth. OUTCOME: Reduced fetal and maternal morbidity and mortality. EVIDENCE: MEDLINE and Cochrane databases were searched using the key words "vacuum" and "birth" as well as "forceps" and "birth" for literature published in English from january 1970 to June 2004. The level of evidence and quality of recommendations made are described using the Evaluation of Evidence from the Canadian Task Force on the Periodic Health Examination. RECOMMENDATIONS: (1) Nonoperative interventions such as one-to-one support, partogram use, oxytocin use, and delayed pushing in women using epidurals will decrease need for operative birth (I-A). (2) Manual rotation may be used alone or in conjunction with instrumental birth with little or no increased risk to the pregnant women or the fetus (III-B). (3) Routine episiotomy is not necessary for an assisted vaginal birth (II-1E). (4) When operative intervention in the second stage of labour is required, the options, risks, and benefits of vacuum, forceps, and Caesarean section much be considered. The choice of intervention needs to be individualized, as one is not clearly safer or more effective than the other (II-B). (5) Failure of the chosen method, vacuum and/or forceps, to achieve delivery of the fetus in a reasonable time should be considered an indication for abandonment of the method (III-C). (6) Adequate clinical experience and appropriate training of the operator are essential to the safe performance of operative deliveries. Hospital credentialing boards should grant privileges for performing these techniques only to an appropriately trained individual who demonstrates adequate skills (III-C). VALIDATION: The Clinical Practice Obstetrics Committee and Executive and Council of the Society of Obstetricians and Gynaecologists of Canada approved these guidelines. PMID- 15779113 TI - Combining parenting and a science career. PMID- 15779112 TI - Abstracts of the American Urological Association Annual Meeting. May 21-26, 2005, San Antonio, Texas, USA. PMID- 15779114 TI - Combining parenting and a science career. PMID- 15779115 TI - Combining parenting and a science career. PMID- 15779116 TI - Crying "Whorf". PMID- 15779117 TI - Edward D. Freis. PMID- 15779118 TI - Believe good things can--and will--happen. PMID- 15779119 TI - Latin American foods lower estrogen levels, maybe breast cancer risk. PMID- 15779121 TI - The cost of late-life depression. PMID- 15779120 TI - By the way, doctor. I take 20 mg of Lipitor and 10 mg of Zetia. My total cholesterol has declined from 177 to 145, my LDL from 133 to 93, but my "good" HDL cholesterol also went down from 46 to 41. Would you consider my low HDL level a large risk factor? If so, is there a regimen that would increase it? PMID- 15779122 TI - Post-tsunami Banda Aceh--on the road to recovery. PMID- 15779123 TI - New HIV/AIDS vaccine enters phase II trials. PMID- 15779124 TI - Edith Silverglied Lisansky Gomberg. PMID- 15779125 TI - Remediation strategies and students' clinical performance. PMID- 15779126 TI - Computer-assisted learning of neurological examination. PMID- 15779127 TI - [125th anniversary of Martin Kirschner]. PMID- 15779128 TI - Perioperative analgesia for knee arthroplasty. PMID- 15779129 TI - Presentation of the Morris F. Collen Award to Clement J. McDonald, MD. PMID- 15779131 TI - A healthier heart, hour by hour. The choices you make throughout your day can add up to longer life. PMID- 15779130 TI - Environmental effects of ozone depletion and its interactions with climate change: progress report, 2004. AB - The complexity of the linkages between ozone depletion, UV-B radiation and climate change has become more apparent. PMID- 15779132 TI - A painkiller is pulled from the market. So what do you do now? PMID- 15779133 TI - Heart-protective drugs: a primer. Back to basics on how they work, side effects, interactions. PMID- 15779134 TI - Medical mystery. A woman with too-thin blood. Why was the patient bleeding? What's her case mean to you? PMID- 15779135 TI - 3 easy-going cardio workouts. The pros and cons of low-impact, high-benefit exercises. PMID- 15779136 TI - Ask the doctors. You've reported that statins may cause damage to the muscles, liver and kidneys. Is the problem temporary or permanent? You also say that a normal CK test indicates no muscle damage. Does that mean that an abnormally high CK level definitely reveals no damage? PMID- 15779137 TI - [Sternoclavicular dislocations: clinical and therapeutic study. A report of 9 cases]. AB - Sterno-clavicular dislocations represent an uncommon pathology in shoulder trauma. The authors describe the anatomical and clinical forms observed between January 1999 and December 2002, and evaluate the treatment process. The retrospective study addressed 9 patients (men only) with an average age of 26.6. Based on the duration of the consulting period, one could distinguish old forms from more recent ones. Diagnosis would help to see anterior forms from posterior forms. Evaluation criteria were: pain, mobility of the shoulder, visibility of the deformation of the sterno-clavicular joint. Six types of anterior dislocations were listed, including 4 recent and 2 old ones. There were 3 recent posterior forms. The anterior recent forms, which were treated in orthopedics, showed a positive evolution. As there was no functional disability, it was wiser to refrain from any treatment of the anterior old forms. The posterior forms, two of them were operated (osteosynthesis) revealed no functional deficit after treatment. However, one of the patients had a broken implant. Scanner facilitated the diagnosis of sterno-clavicular dislocations. The anterior forms, which are more frequent and benign, are different from the posterior forms, which are emergency cases due to the proximity of the aero-digestive and cardiovascular elements. Their surgical stabilization requires techniques related to ligamentoplasty rather than using osteo-synthesis equipment which may move away or break off. PMID- 15779138 TI - [HIV in workplace: knowledges and attitudes of workers in three Senegaleses manufactures]. AB - This study was conducted from may to august 2000 in three senegalese work sites randomly selected among seven which benefited from HIV preventive program. The aims were to evaluate knowledge, attitude and behavior related to HIV infection. A sample of 200 workers randomly chosen were asked about the ways of transmission and methods of protection as well as their risky attitudes and behaviors. Mean age was 41 years, 82.5% were male and 75.5% benefited from high school education; 75.75% had a good level of knowledge on HIV infection, 58% CI [51-65] stated having changed their sexual behavior into abstinence, fidelity to their partner (52.5%) and use of condom (46.5%). This good level of Knowledge and the positive changes in attitude are related to the high rate of alphabetization in our sample, information message diffused by media and preventive programs undertaken in workplace. Information campaigns via media should be maintained and strengthened and preventive programs diffused in all workplaces which will reduce diffusion of HIV in population. PMID- 15779139 TI - [Bronchial hyperreactivity syndrome : a case report following exposure to sulfur dioxide]. AB - A variety of occupational asthma is induced by important exposure to irritants. A case is reported, where le patient developed asthma symptoms like, following huge exposure to sulfur dioxide. All Books criteria were met, except for the metachlorine challenge which was not performed. Nevertheless the bronchial reactivity in the work place, shown by a portable pick flow eased le diagnosis. General practitioners and occupational physicians should be aware of this new medical entity for a better management. PMID- 15779140 TI - Global Tobacco Surveillance System (GTSS): purpose, production, and potential. AB - The World Health Organization (WHO), Centers for Disease Control and Prevention (CDC), and Canadian Public Health Association (CPHA) developed the Global Tobacco Surveillance System (GTSS) to assist all 192 WHO Member States in collecting data on youth and adult tobacco use. The flexible GTSS system includes common data items but allows countries to include important unique information at their discretion. It uses a common survey methodology, similar field procedures for data collection, and similar data management and processing techniques. The GTSS includes collection of data through three surveys: the Global Youth Tobacco Survey (GYTS) for youth, and the Global School Personnel Survey (GSPS) and the Global Health Professional Survey (GHPS) for adults. GTSS data potentially can be applied in four ways. First, countries and research partners can disseminate data through publications, presentations, and an active GTSS web site. Second, countries can use GTSS data to inform politicians about the tobacco problem in their country, leading to new policy decisions to prevent and control tobacco use. Third, GTSS can provide countries with valuable feedback to evaluate and improve Country National Action Plans or develop new plans. Fourth, in response to the WHO FCTC call for countries to use consistent methods and procedures in their surveillance efforts, GTSS offers such consistency in sampling procedures, core questionnaire items, training infield procedures, and analysis of data across all survey sites. The GTSS represents the most comprehensive tobacco surveillance system ever developed and implemented. As an example, this paper describes development of the GYTS and discusses potential uses of the data. Sample data were drawn from 38 sites in 24 countries in the African Region, 82 sites in 35 countries in the Americas Region, 20 sites in 17 countries and the Gaza Strip/West Bank region in the Eastern Mediterranean Region, 25 sites in 22 countries in the European Region, 34 sites in six countries in the Southeast Asia Region, and 25 sites in 14 countries in the Western Pacific Region. PMID- 15779141 TI - Being courtside: your behavior is important. PMID- 15779142 TI - Findings of scientific misconduct. PMID- 15779143 TI - The effect of simulated microgravity on hybridoma cells. AB - The effect of clinostat-simulated microgravity on SP-2/0 and 1D6 hybridoma cells was studied. Clinorotation during 4-5 days at 1.5 rounds per minute decreased dramatically their proliferating capacity: the rotated cells divided less than once while control cells performed 4-5 divisions. They decreased the non-specific adhesion to tissue culture plastic, but increased the number of cell-to-cell contacts. Such phenomenological changes were accompanied with the alterations in pericellular glycosaminoglycans: decreased accumulation of hyaluronic acid and increased accumulation of chondroitin/dermatan-sulfate, as well as with the increase of cytoplasmic Ca2+ concentration. Clinorotation resulted in hybridoma nicotinic receptor desensitization but not down-regulation. In contrast, both the quantity and quality (molecular isoforms, affinity and specificity) of the antibody produced by 1D6 hybridoma cells were not altered by clinorotation. It is concluded that simulated microgravity affected the proliferating and adhesive, but not biosynthetic properties of hybridoma cells. PMID- 15779144 TI - Kinetic measurements of DNA hybridization on an oligonucleotide-immobilized 27 MHz quartz crystal microbalance. AB - A highly sensitive 27-MHz quartz-crystal microbalance, on which a 10-30-mer oligonucleotide was immobilized as a probe molecule, was employed to detect hybridization of complementary oligonucleotides in aqueous solution. From frequency decreases (mass increases due to the hybridization) with passage of time, kinetic parameters such as association constants (K(a)) and binding and dissociation rate constants (k(1) and k(-1)) could be obtained, as well as binding (hybridization) amount at the nanogram level (delta m). Kinetic studies were carried out by changing various parameters: (i) the immobilization method of a probe oligonucleotide on Au electrode, (ii) number of mismatching bases in sequences of target oligonucleotides, (iii) length of both probe and target oligonucleotides, (iv) hybridization temperature, and (v) ionic strength in solution. The obtained results were compared with those obtained by a surface plasmon resonance method using a BIAcore system. PMID- 15779145 TI - Funding health advocacy. PMID- 15779146 TI - Aquatic injury. Near drowning. PMID- 15779147 TI - Autologous preparations rich in growth factors promote proliferation and induce VEGF and HGF production by human tendon cells in culture. AB - Blood platelets become activated and aggregate at the site of vessel injury. Upon activation by thrombin, platelets release storage pools of proteins and growth factors (GFs), including those involved in tissue repair. Our goal was to evaluate the potential beneficial effect of proteins released from platelet-rich clots on tendon healing. PDGF, TGF-beta-1, IGF-I, HGF, VEGF and EGF were measured in human platelet-poor plasma (PPP) and in the releasates collected from either platelet-poor or platelet-rich clots prepared in vitro. We then studied the effects of the releasates on human tendon cells in culture. Releasates from both platelet-rich and platelet-poor clots stimulated tendon cell proliferation, in contrast to un-clotted PPP. The mitogenic activity of the supernatants was not decreased by the thrombin inhibitor, hirudin. Cultured tendon cells synthesise VEGF and HGF in the presence of PPP-clots and PRP-clot releasates, thus the synthesised amount was significantly higher with supernatants from platelet-rich clots than supernatants from a platelet-poor clot (p < 0.05). These results suggest that administering autologous platelet-rich clots may be beneficial to the treatment of tendon injuries by inducing cell proliferation and promoting the synthesis of angiogenic factors during the healing process. PMID- 15779148 TI - OHAfrica Lesotho Initiative--for hope, health and healing. PMID- 15779149 TI - Building momentum. PMID- 15779150 TI - Building momentum. PMID- 15779151 TI - Manual ventilation and risk of barotrauma: primum non nocere. PMID- 15779152 TI - Paraquat poisoning. PMID- 15779153 TI - Methemoglobinemia and pulse oximetry. PMID- 15779154 TI - Dynamic hyperinflation, intrinsic positive end-expiratory pressure, and respiratory rate. PMID- 15779155 TI - Expiratory rib-cage compression, airway suctioning, and atelectasis. PMID- 15779156 TI - [Promoting improvement of liver transplantation in China]. PMID- 15779157 TI - Quarterly index. Use of health services and SARS. PMID- 15779158 TI - [Trends of liver transplantation]. PMID- 15779159 TI - How to pretend that correlated variables are independent by using difference observations. AB - In many areas of data modeling, observations at different locations (e.g.,time frames or pixel locations) are augmented by differences of nea r by observations (e.g., delta features in speech recognition, Gabor jets in image analysis). These augmented observations are then often modeled as being independent. How can this make sense?We provide two interpretations,showing (1) that the likelihood of data generated from an auto regressive process can be computed in terms of "independent" augmented observations and (2) that the augmented observations can be given a coherent treatment in terms of the products of experts model (Hinton, 1999). PMID- 15779160 TI - Convergence of the IRWLS Procedure to the Support Vector Machine Solution. AB - An iterative reweighted least squares (IRWLS) procedure recently proposed is shown to converge to the support vector machine solution. The convergence to a stationary point is ensured by modifying the original IRWLS procedure. PMID- 15779161 TI - [Antidiabetic activity of Zizyphus mauritiana Lam (Rhamnaceae)]. AB - In traditonnal medicine, several plants were used in diabetes treatment. Among them, the aqueous extract from the leaves of Zizyphus mauritiana Lam (Rhamnaceae) was studied. The extract was administrated per os to Wistar rats made diabetics either temporarily by oral glucose tolerance test (first case) or definetely by subcutaneous injection of alloxan (second case). It was observed a striking decrease of the hyperglycemic arrow (p < 0.05) in the first case, with 300 mg/kg administrated 90 minutes before starting the test. In the second case, the results obtained with a dose of 300 mg/kg once or twice a day were identical as those with glibenclamide at 0.2 mg/kg per day. So, the antidiabetic activity of Zizyphus mauritiana Lam was experimentally born out but it has to be standardized for common use. PMID- 15779162 TI - [Chronic cor pulmonale: a study of 34 cases in the Dakar University Hospital Center Cardiology Department]. AB - The authors report the results of a study on patients with chronic cor pulmonale hospitalized in the Departement of Cardiology (Dakar), from 1990 to 1998. The hospital prevalence is 0.9% with a male predominance (73.5%). The clinical signs were dominated by dyspnea and cough. The right heart failure is noted in 85.29% of cases. As far as the biological factor is concerned, it exists an polycythemia about 75% of cases, hypoxia (88.8%), hypercapnia (55.5%) and a respiratory acidoses (55.5%). The spirometry showed in 66% of cases, a mixed syndrom with obstructive predominance. The echocardiography showed in all cases right heart dilatation with pulmonary hypertension. The etiological factors are dominated by tobacco (21 cases). The hospital evolution has been favorable in 19 cases underoxygentherapy, lowdosediuretic, expectorant, bronchodilatators and salt restriction. However, 14 patients have died. PMID- 15779163 TI - [Inflammatory ointment from shea butter and hydro-alcoholic extract of Khaya senegalensis barks (Cailcederat)]. AB - In a former study, it was proved that the alcoholic solution of hydro-alcoholic extract of Khaya senegalensis barks had an anti-inflammatory activity on animals after a local application. In this work, ointments made from the same extract and three different excipients (vaseline, lanoline and shea butter (crude and refined)) have been prepared and tested by the method of the croton oil inhibited ear oedema. Results showed inhibition percentages of the ear oedema of 58.8%, 66.7% and 75.4% when the hydro-alcoholic extract was tested at respective doses of 1%, 2% and 3% in shea butter. The two other excipients, (vaceline and Lanoline) tested at the dose of 3% showed between 52% and 58% of inhibitions. The interest of this study was to demonstrate the possibility to maintain the anti inflammatory activity of Khaya senegalensis barks by using them in a galenic form, easy to prepare and which is, in addition, more adapted than the extract to possible clinical trials. PMID- 15779165 TI - [Mango leaves (Manguifera indica, Anacardiacea) used as biomarker of pollution by pp'Dichlorodiphenyltrichloroethane (pp'DDT)]. AB - Dichlorodiphenyltrichloroethane (DDT), organochlorine pesticide, byanother way cumulative pesticide is banned in mostdeveloped countries. Whatever, it is still used in many countries in the Third World. This work consist to check whether this chemical compound is still used in Senegal. The biomarker of pollution by this pesticide for this research was mango leaves (mango tree is widespread in Senegal) open to accumulate halogenated hydrocarbons. Leaves sample were taken in many sites in the centerline of Dakar-Thies (Senegal), then analysed by gas chromatography. The results allowed to note the presence of DDT and its metabolites [(dichlorodiphenyltrichloroethylen (DDE), (dichlorodiphenyltrichloroethane (DDD)] in most of samples. The comparison of DDT content in relation to those of the principal metabolite allowed to make the difference according to sample sites, an old contamination of an utilisation more or less recent of this pesticide. From those results, we can conclude that DDT, typical pesticide by its big persistence is still used in some spherical zones in this country, in particular in farming gardening zones. PMID- 15779164 TI - [Specific antibodies to Plasmodium falciparum antigens in immune patients: IV- Cross-sectional analysis of anti-P. falciparum Ab responses in a mesoendemic zone of transmission in Sine-Saloum region]. AB - We report here a comparative study of specific antibody responses to blood stages of P. falciparum in 100 individuals aged from 3 to 76 years, living in the village of Ndiop (an area where parasite transmission is approximately 20 infective bites per year). We have used 2 antigen preparations: crude merozoites and infected erythrocyte ghosts (GRp). High specific antibody responses were found in the villagers' sera, with IgG levels being significantly higher than those directed to the whole P. falciparum schizont Ag preparation; the reverse was found for IgM responses. An imbalanced IgG subclass Ab response was evidenced towards cytophilic IgG1+/gG3+ responses rather than to IgG2+/lgG4+. Mean levels of antibodies increased significantly with age and a strong relationship was evidenced in individual levels of Ab responses towards all the Ag studied. Investigation of the morbidity, recorded as the number of confirmed clinical accesses, underlined a significant inverse relationship only with the levels of IgG to GRp. Taken together, these results proved usefullness of crude P. falciparum Ags for immuno-epidemiological related studies and underlined GRp as a particular target of Ab responses. PMID- 15779166 TI - [Role of nitric oxide in diaphragmatic dysfunction genesis during sepsis in rats]. AB - Nitric oxide (NO) is a vasodilator agent that is cytotoxic and negatively inotropic in the heart. More recently, it has been shown that during sepsis there is a high amount of NO production by a NO synthase (NOS) that is inducible by cytokines. The aim of this study was to investigate the role of NO in the genesis of diaphragmatic dysfunction during sepsis. Rats were inoculated i.p. injection with 10 mg/kg of Escherichia coil endotoxin (E animals) or saline (C animals). Six hours after endotoxin or saline inoculation, diaphragmatic force and muscularc GMP (Cyclic guanosine monophosphate) were assessed by in vitro force frequency curves and ELISA method, respectively. As compared to C animals, E animals showed a significant decrease in diaphragmatic force for all the frequencies of stimulation (p < 0.01). This reduction was associated with a significant increase in muscular cGMP. Inhibition of NO synthesis in E animals with either dexamethasone (4 mg/kg IV, 45 min before endotoxin or saline) or NG monomethyl-L-arginine (L-NMMA, 8 mg/kg IV, 90 min after endotoxin or saline) prevented the effects of endotoxin. However, no modification was seen with NG monomethyl-D-arginine (D-NMMA), a molecule which does not inhibit NO synthesis. Administration of dexamethasone or L-NMMA in C animals did not induce any significant change in diaphragmatic force, and cGMP ratio. We conclude that NO has a contributive role in diaphragmatic dysfunction during Escherichia coli induced sepsis in rats. PMID- 15779168 TI - [Aflatoxin contamination level in artisanal and industrial peanut butter food in Dakar (Senegal)]. AB - Aflatoxins are mycotoxins produced by some strains of fungus (Aspergillus) which develop in peanut seeds. Peanut oil and past are very used up in Senegal, then the aflatoxin poisoning risk is very actual. The aim of this study was to determinate the aflatoxin level in artisanal and industrial peanut pastry food from Dakar (Senegal). High Performance Liquid Chromatography (HPLC) analysis of the different samples showed that the most contaminated by aflatoxins are artisanal pastry sold in different market of Dakar (Senegal). Indeed, 40% of these samples contained mean values of aflatoxin B1 (the most dangerous) widely over allowable EEC specifications (5ppb). Furthermore, most of industrial and domestic peanut pastry were cleaned and could be consumed without risk. PMID- 15779167 TI - [Peripartum heart failure: the underestimated role of frequent diseases in the Sudan-Sahelian area]. AB - Peripartum cardiomyopathy is rare in developped countries, but still frequent in Africa. It is defined as a heart failure occurring during peripartum, without any underlying etiology. Authors present 3 cases showing that heart failure before or after delivery may be due to causes which are frequent in the Sahelian area but generally misdiagnosed. Anemia, hypertension and rheumatic fever were the causes of heart failure in these 3 patients, but they were not apparent when the initial diagnosis was made. These observations emphasize that, despite the complex hypothesis trying to explain heart failure during the peripartum period, one should think about some frequent causes which can be misdiagnosed because of the pregnant state or the heart failure itself. PMID- 15779169 TI - [Value of post coital and cytological sperm tests in conjugal sterility in Senegal]. AB - Conjugal infertility in Senegal is surrounded with accusatory believes most oftenly unfavourable to woman who is generally incriminated and, in somes cases, repudied. This study is an account of a research work carried out on conjugal infertility causes analysis, using post coital and cytospermiologic tests. It aims to help our populations to better appreciate the conjugal infertility by giving them elements for an objective appraisal of this flail, particulary about the mutual responsabilities of the two partners. In a first stage, we run post coital tests with 4636 Negro-African infertile couples living in Senegal, at the Clinical Cytology, Cytogenetics, and Reproductive Biology Laboratory of Aristide Le Dantec Hospital in a period dated from 1983 to 1996. Post coital tests were abnormal for 3838 couples and 3150 of them had anomalies incriminating the husbands. In a second stage, post coital tests were suggested to these 3150 husbands, but only 2842 accepted the post coital test suggestion. The others refused it for sociocultural reasons. It stooks out of this study that: the responsability in the conjugal sterility is mixed and shared by both partners in 43.78% cases, the masculine responsability (38.30%) ruled the feminine one (17.92%) contrary to local believes, almost one husband out of four (23.89%) presented azoospermy, the feminine responsability is ruled by infection factors that represented half of the glair abnormalities (50%). These results contradict the present local mentalities and believes which accuse principally the woman. PMID- 15779171 TI - [Surgical aspects of pulmonary aspergilloma in 24 patients]. AB - From January 1991 to July 1998, 24 patients were operated for pulmonary aspergilloma. All the patients presented previous tuberculosis treated. Hemoptysis had been the revealing elements of the illness in 80% of cases. Treatment was by resection in all the cases: 13 pneumonectomy (54%) and 11 partial resections (46%). The procedures were usually difficult and hemorrhagic. The global mortality was 20%, can be explained by the high number of major resections and the unfavourable general state of health of our patients. Post operative complications are mainly due to secondary empyemas encountered in 15% of the surviving ones. The surgical resection of pulmonary aspergilloma is still difficult and risky for extended underlying parenchymal disease. Minima procedures such as speleotomy and intra cavitary aspiration technique must be indicated in patients with very poor general condition. PMID- 15779170 TI - [Assessment of the level of urinary iodine deficiency in children of Senegalese central regions]. AB - Iodine deficiency disorders (IDD) are a major public health problem in Senegal, where strategies of salt iodization were adopted in the southern and eastern regions. The aim of this study led in four districts (Koungheul, Bambey, Mekhe and Kebemer), was to estimate by a questionnaire, the women knowleges, attitudes and practices (KAP) concerning IDD, and to measure children urinary iodine excretion by the Sandell-Kolthoff method to assess a potential deficiency. Six hundred ninety eight households were selected covering 1336 women (age 15 to 49 years) and 400 children (age 6 to 12 years). Sixty three per cent of the women knew the goiter, 89% of them considered that it was a disease and only 0.6% knew the role of iodized salt in the treatment. On the other hand, 20% of the children presented a normal range of urinary iodine excretion superior to 100 microg/l, the deficiency was light (50 to 100 microg/l) in 38% of the children, moderate (25 to 50 microg/l) in 27% and severe (< 25 microg/l) in 15% of them. These results show that other Senegalese regions are concerned by iodine deficiency disorders and need information, education and iodine supplementation programmes, notably for children and young women. PMID- 15779172 TI - [Early post-operative arrhythmias in endomyocardial fibrosis or chronic parietal endocarditis in 84 cases studied at the Cardiology Institute of Abidjan]. AB - The authors have studied rhythmic events happened during early post-operative period in 84 operated patients for endomyocardial fibrosis, synonym nowadays to chronic parietal endocarditis, in the Institute of Cardiology of Abidjan (Cote d'lvoire), from January 1977 to July 1991. The mean age has been 15.6 +/- 43 years old. Endocardectomy was left in 25 cases, right in 32 and at last bilateral in the 19 others. The surgical way has been a left and/or right atriotomy. Mitral and/ ortricuspide valvular surgery has been always realised. The absolute prevalence of patients having presented one or several arrhythmias, atrial tachycardia and premature ventricular beats. The favouring factors have been the period of exclusion of the aortic root from circulation and number of endomyocardectomy. Atrial fibrillation has been related to auricular volume. Most of nodal arrhythmias have appeared with surgery. Endomyocardial fibrosis surgery has been greatful in rhythmic plan in 16% of patients. Early post-operative arrhythmias have been responsible forone case of the death of the operated patients and they have been present in seven other cases of the death. PMID- 15779174 TI - [Prevalence of the use of skin bleaching cosmetics in two areas in Dakar (Senegal)]. AB - The use of skin bleaching cosmetics is an increasingly widespread phenomenon in Africa. If the negative sanitary effects of this practice are today well known, reliable statistics on the importance of the problem within the global population are rare. Our study, conducted in two popular areas in Dakar, Usine bene taly and Usine Niary Taly, had 2 objectives: to determine the prevalence of the use of skin bleaching cosmetics in the studied zones, to describe the prevalence according to the following variables: age, matrimonial situation, level and/or type of instruction, job, type of agents used. For that purpose, we conducted a transverse study on a representative sample of 600 women from 15 to 55 years old. This study reveals a prevalence of the use of skin bleaching cosmetics of 67.2%. The agents found are hydroquinone derivatives (61%), topical corticosteroids (37%) and agents from unknown origin (2%). The use of skin bleaching cosmetics is more important among young women between 30 to 44 years (72.5%), married (72.2%), analphabets (75%), working (77.6%). Studies conducted in Mali and Togo gave comparable results, which give perspectives for a prevention based on education an awareness. PMID- 15779173 TI - [Friedreich's disease in the department of neurology in Dakar]. AB - A retrospective study of Friedreich's ataxia was conducted from january 1960 to december 1997. Eighten cases had been collected. Friedreich's ataxia was the second disorder after Pierre Marie's ataxia among inherited ataxia. Parental consanguinity has been found in 27.77% of cases. The sex-ratio was 2.6 for males. 66,67% of Friedreich's ataxia cases began after 25 years. Clinical signs were: cerebellar ataxia, sensitive disorders in 70%, pyramidal syndrom and cardiomyopathy in 22.22%, bones dysmorphy in 50% of cases. Electromyography indicated severe axonopathy of members with decreased somesthesic potentials in six cases. Glycaemia was normal in all cases. Clinical future presented wide variation between one and fourth years old. PMID- 15779175 TI - [Prevalence and clinical aspects of puerperal mental disorders at the University Hospital Center of Dakar]. AB - This is a contribution to the survey on psychiatric disorders of puerperality. We are particularly interested in the clinical signs and the ethiopathogenical facts of psychiatric disorders of the post-partum which appear-in our point of view-to be the main point on which we could probably act to reduce the effect of the disease. It is a retrospective survey going from January, 1st 1992 to December, 31st 1996. Here are included all the female patients whose age is more than 15, hospitalised during survey period and who presented a mental pathology linked with puerperality in accordance with diagnosis criterions of World Health Organization. In 1 627 hospitalisations, there were 658 women among whom 52 presented puerperality psychical disorders, that's meaning 7.9% of the female patients hospitalised in psychiatry, with an average age of 24.5 years. Concerning the classification of the diseases, the acute delirious psychosis and the depressive disorders are more important with respectively 56% and 19% of the sample. 92% of the women gave birth by the vaginal normal way and 61.5% presented disorders in the first week following their delivery. The married women are 94%. Concerning their occupations, 98% are house keepers. 52% are primiparas and 48% are multiparas. Psychical disorders of puerperality are frequent in Dakar. Acute psychosis and mood disorders are the main clinical aspects. The management of the future mother to her new roles by health education and prenatal check-ups in health centers by skilled and appropriate personnal is necessary. PMID- 15779176 TI - [Epidemiological, clinical and progressive aspects of neurological manifestations associated with retroviral infections: eleven year retrospective study]. AB - Through a cohort of 93 neuroaids which has been diagnosed at Dakar in our Neurology Department, the authors evaluated the hospital prevalence of retrovirus, detected socio-demographic factors, related AIDS outline the mean neurological picture and try to correlate survival and neurological involvement of these patients. Among 1151 patients who got retroviral blood test, 93 were seropositive (8.1%). On these repartitions 36 were females (38.7%) and 57 males (61.3%). The age goes from 19 to 76 years old. 45 patients (48.4%) have been found positive for HIV-1 blood test, 21 patients (22.6%) for HIV-2 blood test, 11 patients (11.8%) for both HIV2, 11 patients (11.8%) for HTLV1, 3 patients (3.2%) for both HIV-1 and HTLV1, and 2 patients (2.2%) for both HIV-2 and HTLV1. In our study the transmission of AIDS occur mainly through heterosexual inter course and multiple parternship is a high risk group. The central nervous system deseases represented 68.8% of cases. The pathology were dominated by stroke, myelopathies, meningoencephalotis and spinal cord compression. The peripheral nervous system desease were found in 7.5% of cases. The peripheral facial paralysis occupied the first place in HIV infections of peripheral nervous system deseases (57.1% of cases). When neurological involvement set up the letality is higher for HIV-1 (57% of global letality) and for central system nervous involvement (76.2%). PMID- 15779177 TI - [Aetiological aspects of neurological diseases in Dakar: follow-up after 10 years (1986-1995)]. AB - Because of the apparition of new risk factors and numerous progresses in investigation methods, authors take stock of neurological diseases of patients admitted in Clinique Neurologique of Fann during the last ten years. The 8539 cases repartition is as followed: 4736 males and 3803 female. Their ages ranged between 3 and 80 years with 2130 deaths corresponding to 24.94%. Annual mean of admission is 853 +/- 42 cases. Aetiological data are: stroke (3910 cases 45.78% with 60.56% of death). The following is peripheral neuropathieswith 714 cases(8.61%) and the spinal cord compression with 692 cases (8.10%). Comparision of frequencies of different aetiologies with those of previous studies shows that the small capacity of the neurological department is a restricting factor for the entry frequencies of neurological patients; the principal pathologies got small variations from one decade to another. In the opposite, some aetiologies have disappeared (neurological syphilitis, trypanosomiasis, neurocyticercosis, cerebral mycosis). Some eatiologies remain unprecised (infectious diseases, peripheral neuropathies, degenerative diseases). PMID- 15779178 TI - [Breast cancer and pregnancy: about 60 cases collected at the Cancer Institut of Dakar]. AB - The purpose of this retrospective study of 60 breast carcinoma during pregnancy collected at the Cancer Institut of Dakar from 1962 to 1998 was to determine the epidemiology, the clinical appearance and to discuss the pathogeny, the diagnosis, the prognosis and the treatment. The mean of age of patients was 34.5 years. The number of parity was 5 and the first pregnancy at 20 years. The authors arrived at diagnosis in the post partum in 28 cases (47.45%), in the first trimester in 8 cases (13.55%), in the midtrimester in 13 cases (22.03%) and in the third trimester in 11 cases (18.64%). The polychemotherapy preceded the surgery in 30 cases and succeded in 16 cases. The hormonotherapy with Tamoxifen prescribed in 17 cases after the delivery. The mastectomy and axillary lymphadenectomy was realised in 26 cases in the post partum. The adjuvant radiotherapy was done in 12 cases. The mortality was 31% and the global survival at 2 years, 34.6%. PMID- 15779179 TI - [Actual experiences of menopause in 342 women]. AB - The objective of this study is to assess the actual experiences of menopause in menopaused women without hormone replacement therapy. A cross-sectional study was conducted among 342 women who had natural menopause. The target population was recruited from subjects attending the Departement of Gynecologie-Obstetrique du CHU TOKOIN from November 1996 to October 1997. All women investigated presented clinical signs related to menopause. 51.5% found its symptoms embarrassing. 53.8% of the women enjoyed their menopause. The difference between the two groups of women having accepted their menopausal status or not, where the menstrual period were embarrassing or not (p = 0.0001) and were the climateric symptoms were embarrassing or not (p = 0.0001) were significant. Agreement to undertake a hormone replacement therapy were given by 29.8% of women. In spite of the climateric symptoms, menopaused women hardly accept their status and few were available for a hormone replacement therapy. PMID- 15779180 TI - [Caries and gingivitis study among preschool children (2-5 years) of the region of Ziguinchor in Senegal. Strategies of prevention]. AB - We studied the preschool (2-5 years) group of child's oral health status in Ziguinchor, the southern regions of Senegal. Our objective was to make the inventory of fixtures dealing with dental caries, gingivitis and tartar among the children of this age group. It's a transversal study based on a clinical examination of a sample size of 168 children collected by stratified random sampling in 12 neursey schools in the region. We used carious (dmf/t), gingival and plaque indices to estimate, orodental status of the population constituting the sample. In the lump, 60.71% of the children have a natural history of dental caries with a mean dmf/t of 2.47 and gingivitis and or tartar are observed in 42.86% of the children. Even children less than 3 years are affected. None age may no longer be free from these diseases. That lead us to suggest to set up in this regional area a preventive program involving mainling the parents and the educators and based on the oral health education and the optimal use of dental products with fluoride since the drinking water in the region is poor in fluoride. The aim of this program would be to reduce the prevalence of the diseases in the proportions indicated by the world health organization and even eradicate these diseases among the prescolar children during the coming years as it's going on actually in the developed countries. PMID- 15779181 TI - [Tuberculosis spondylodiscitis in a neurological service in Nouakchott]. AB - It is about a retrospective study dealing with the place of tuberculous spondylitis with discitis among medullary wounds hospitalized in our service from January 1995 to June 1998 and their medical care. Twenty reports have been done making: 30.30% of our medullary wounds. We have noted a male prevalence and precocity. The flaccido-spastic paraplegia with sensitive level prevails the clinical chart (45%) this in correlation with thoracic and lumbar localization predominance (75%) at the radiography. All patients have had for their benefit an anti-tuberculous treatment and eleven among them had also tetracosactide (synactene retard). This has allowed us to observe a quick recovery of the motor deficit for nine patients: 81.8% of the patients who have had the therapeutic association; 45% of the whole. No patient had an operation. PMID- 15779182 TI - [HBs antigen carrier state in pregnant women in Bobo Dioulasso (Burkina Faso)]. AB - The aim of this study was to evaluate the prevalence rate and the risk factors for the carriage of hepatitis B markers in pregnant women in Bobo Dioulasso, Burkina Faso. Out of 917 pregnant women recruited during antenatal care, 98 (10.7%) were HBs antigen positive. Among these ones, 18.2% carded HBe antigen, 66.7% antiHBe antibodies and 95.6% antiHBc antibodies. Two risk factors were identified: maternal age of 23 and 28 (RR = 2.33, chi2 =12.21, p = 0.005) and widowage (Fisher test RR = 6.43, p = 0.0016). This high prevalence of HBs antigen calls for systematic screening for hepatitis B during antenatal care along with an immunization policy toward women of reproductive age and newborns. PMID- 15779183 TI - [Giant ovarian cyst and pregnancy (a case report)]. AB - We report a case of giant ovarian cyst in pregnancy of 23 weeks of amenorrhea revealed by digestive and respiratory compression. As the absence of ultrasonographic and biologic malignancy criterias, several punctures of the cyst were done permitting the evolution of pregnancy until the preterm period. An annexectomy done in the immediate post-partum permitted to avoid the frequent mechanical accidents during this period. From this case, we discuss clinical and therapeutic problems of this association. PMID- 15779184 TI - [Shigella bacteraemia: a report of two cases observed in Dakar]. AB - Shigellosis is usually a self-limiting enteric disease, rarely accompanied by extra-intestinal manifestations. In Senegal, shigella bacteraemia is poorly documented. The authors report two retrospective cases of Shigella dysenteriae 1 bacteraemia in an infant aged 12 months and a 32 years-old adult admitted at the Infectious Diseases Clinic in Dakar. In both cases, a fatal outcome was observed, in relation to complications: endotoxic shock in the infant, severe intestinal haemorrhage in the adult. PMID- 15779185 TI - [Lymph node and cutaneous tuberculosis revealed by a pyloroduodenal stenosis: a case report]. AB - Lymphadenitis is the most frequent form of extrapulmonary tuberculosis. In immunocompetent patients, its clinical presentation is usually superficial, less frequently intra-abdominal located, and exceptionnally symptomatic. We report a case of scrofuloderma characterized by the lymph nodes extent of spread which led to a pyloro-duodenal stenosis. An 18 years old immunocompetend man is admitted in the internal medicine department, presenting a pyloroduodenal stenosis syndrome, fever, weight loss, and ulcerated axillary and cervical lymph nodes. The upper gastrointestinal endoscopy and ultrasound examination reveal the extrinsic duodenal compression by large numerous lymph node. A biopsy confirms the tuberculosis disease for which no other localisation was detected. Despite of the diagnostic and therapeutic delay, which explains the expanse and depth of the lesions, the antituberculosis therapy was effective on the digestive symptom and cleared all the cutaneous manifestations. PMID- 15779186 TI - [Biventricular thrombus complicating peripartum cardiomyopathy. A case report]. AB - The authors report a case of a biventricular thrombus complicating peripartum cardiomyopathy in a 38 years old female. The diagnosis was done by bidimensionnal transthoracic echocardiography which showed 2 thrombi in the apical region of the right ventricle and in the anterior and lateral wall of the left ventricle. With treatment associating salt restriction, diuretics and angiotensin-converting enzyme- inhibitors, the evolution was good: the thrombi disappeared at the first month for the left ventricular thrombi and after 45 days for the rignt ventricular thrombi. The patient didn't experience any embolic complication. PMID- 15779187 TI - [Delayed root canal therapy. Data on the frequency and complications]. AB - This retrospective study of 118 teeth receiving root canal treatment was conducted to determine the frequence and complications of delayed root canal treatment. Teeth were categorised into prompt treatment group and delayed treatment group according to the length time of treatment; prompt treatment group was defined as a group where root canal treatment was achieved within a period of 4 months or less, and delayed treatments group, a group where period of treatment was greater than 4 months or where canals were never obturated. Two types of complications were studied: postobturation pain and fracture involving tooth extration. The results of this study have shown that 34.5% of treatments were delayed. In this group we found also a higher rate of fractures involving tooth extraction, but the difference was not significant. The results have also shown that disappearance of pain was the essential factors in the frequence of delayed treatment, but the dental chair breakdown had some responsability. As well as in the two groups, the first reason of extractions was fracture and not pain. These results emphasize the necessity of focusing the patient's motivation on the risk of losing the tooth. PMID- 15779188 TI - [The pathology of the peritoneo-vaginal process in the young males: clinical and therapeutical aspects in 160 cases]. AB - The objective of this work was to describe the clinical and therapeutical aspects of pathology of the peritoneo-vaginal process. We have performed a retrospective study including 160 patients operated between January 1990 up to December 1996. Mean age at diagnosis was 8 years, ranged from 1 month to 13 years old. All patients were male. The abnormality was located in the right side in 60% of cases and was bilateral in 6.7% of cases. The main clinical features were scrotal mass (81%) and scrotal pain (13.46%). The diagnosis was made at birth only in 20% of cases. A maldescended testis was associated in 7.5% of cases. A groin incision have been used in 91.25% of patients. The average hospital stay after surgery was 1 day. Thus, the pathology of the peritoneo-vaginal process is common and apparently banal. Need for treatment through a groin incision owing to the possibility of associated maldescended testis. PMID- 15779189 TI - Speaking of good health. PMID- 15779190 TI - Jean Wishnick King: a true "slot" woman. PMID- 15779191 TI - Jean. PMID- 15779192 TI - What to do about a surprise BME investigation. PMID- 15779193 TI - Now more than ever..your profession needs you. PMID- 15779194 TI - Old questions, new playing field. PMID- 15779195 TI - Save yourself first: parenting an addicted child. PMID- 15779196 TI - Zoledronic acid-induced severe hypocalcaemia in a prostate cancer patient with extensive osteoblastic bone metastases. AB - A 74-year old white man was undergoing treatment with palliative chemotherapy for Stage IV Prostate Adenocarcinoma with multiple osteoblastic metastases. He was started on intravenous Zoledronic acid for reduction of bone pain and prevention of skeletal complication from multiple bone metastases. Four days after intravenous Zoledronic acid, the patient presented to emergency room with complaints of carpopedal spasm and bronchospasm. On admission, serum calcium and albumin was 3.5 mg/dl and 3.7 g/dl respectively. QT interval was more than 500 m sec. The patient was started on intravenous calcium gluconate. He received 174 gm of intravenous calcium for total of 12 days. His serum calcium returned to a normal range in three months with Vitamin D and calcium supplement. Before starting Zoledronic acid therapy, the patient's serum calcium level was 6.9 mg/dl. PMID- 15779197 TI - Physician management of death certificates. PMID- 15779198 TI - National doctors' day. PMID- 15779200 TI - Half way there. PMID- 15779201 TI - Will it deliver? PMID- 15779202 TI - Mad boy. Interview by Frank Bangay. PMID- 15779203 TI - Virtual circle. PMID- 15779204 TI - Kiss it XX better. PMID- 15779205 TI - Neglect, not smacking, is the public health issue. To conceive and leave should be against the law. PMID- 15779206 TI - Under current law the right to change the 'nearest relative' is weighted in favour of the psychiatrists. PMID- 15779207 TI - These fiddly Carer's allowance rules were designed to put the mean into means tested. PMID- 15779208 TI - Stop the juggernaught. PMID- 15779209 TI - Reaching out to women. PMID- 15779210 TI - Even elders get the blues. PMID- 15779211 TI - We know what we want. PMID- 15779212 TI - This life. PMID- 15779213 TI - The strategic futures initiative. PMID- 15779215 TI - The soft side. AB - For true communication between dentist and patient, a dentist must develop behavioral skills (or "soft" skills) in addition to clinical skills. These skills should be utilized at all times but particularly during a patient's initial visit. Educating the patient is mandatory and the dentist should never offer solutions to problems that the patient cannot see, feel, or understand. If these ideas are kept paramount at all times they will become normal operating procedure and not seem artificial. Caring about the patient and remaining committed to technical excellence will go a long way toward the delivery of quality care. PMID- 15779214 TI - Aloe vera gel: update for dentistry. PMID- 15779216 TI - Transparent truthfulness. PMID- 15779217 TI - Hula hoops, pet rocks, and low-carbohydrate diets. PMID- 15779218 TI - Rehabilitation of a fearful dental patient with oral sedation: utilizing the incremental oral administration technique. AB - The treatment of fearful or anxious patients presents a myriad of problems for the dentist. In-office sedation using oral (enteral) medications is an effective means of increasing patient tolerance of invasive dental procedures. The incremental oral administration technique is a protocol that can be utilized to treat fearful or anxious patients. A case is presented in which this technique was used as an adjunct to the rehabilitation of a debilitated mouth. PMID- 15779219 TI - Effects of sports drinks and other beverages on dental enamel. AB - A high percentage of people consume soft drinks that contain sugar or artificial sweeteners, flavorings, and various additives. The popularity of sports (energy) drinks is growing and this pilot study compares enamel dissolution in these and a variety of other beverages. Enamel blocks (approximately 7.0 x 5.0 x 2.5 mm) were sectioned from sound extracted human premolars and molars and measured, weighed, and immersed in the selected beverages for a total of 14 days. The pH of all beverages was measured. The enamel sections were weighed at regular intervals throughout the immersion period with the solutions being changed daily; all studies were performed in duplicate. The data were subjected to one-way ANOVA with post hoc Scheffe testing. Enamel dissolution occurred in all of the tested beverages, with far greater attack occurring in flavored and energy (sports) drinks than previously noted for water and cola drinks. No correlation was found between enamel dissolution and beverage pH. Non-cola drinks, commercial lemonades, and energy/sports drinks showed the most aggressive dissolution effect on dental enamel. Reduced residence times of beverages in the mouth by salivary clearance or rinsing would appear to be beneficial. PMID- 15779220 TI - Antimicrobial susceptibility of air-dispersed microorganisms in dental settings. AB - To determine the number and the susceptibility of microorganisms collected in a clinical environment against the antimicrobial agents used commonly in dentistry, petri dishes containing trypticase soy agar were exposed to air in different sites of a multi-chair dental clinic before, during, and after multiple clinical procedures and incubated for 24 hours under aerobic conditions. Colonies were identified by Gram stain technique and biochemical tests. Commercial paper disks containing widely prescribed antimicrobial agents (beta-lactams, macrolides, and clindamycin) were used to perform the antimicrobial susceptibility tests. The groups (colony forming units = cfu/m2/min) were submitted to the Kruskal-Wallis test (alpha = 5.0%), considering different clinical situations and environmental sites. During clinical procedures, the number of microorganisms increased (p < 0.05). This study highlights the need for established strategies to prevent resistant bacterial strains from emerging in dental settings. PMID- 15779221 TI - Shear bond strengths of three different resin luting cements to bovine teeth. AB - This study evaluated the shear bond strengths of dentin/porcelain and enamel/ porcelain specimens bonded with three different dual-cure cements. The specimens were prepared using the heat-pressed ceramic technique. Three different dual-cure cements were used to bond samples to previously prepared bovine enamel and dentin surfaces. The specimens were thermocycled and tested in shear until failure. The shearbond data was subjected to one-way ANOVA, followed by Duncan's multiple range test at a significance level of p < 0.05. The shear bond strengths of Variolink II were significantly higher than those of Rely-X ARC and Panavia F for enamel and dentin (p < 0.05). Bond strenghts in enamel were higher than those of dentin in all materials (p < 0.05). PMID- 15779222 TI - Proximal caries detection: Sirona Sidexis versus Kodak Ektaspeed Plus. AB - This study compared the accuracy of intraoral film and a charge-coupled device (CCD) receptor for proximal caries detection. Four observers evaluated images of the proximal surfaces of 40 extracted posterior teeth. The presence or absence of caries was scored using a five-point confidence scale. The actual status of each surface was determined from ground section histology. Responses were evaluated by means of receiver operating characteristic (ROC) analysis. Areas under ROC curves (Az) were assessed through a paired t-test. The performance of the CCD-based intraoral sensor was not different statistically from Ektaspeed Plus film in detecting proximal caries. PMID- 15779223 TI - Comparison of the degree of pulpal anesthesia achieved with the intraosseous injection and infiltration injection using 2% lidocaine with 1:100,000 epinephrine. AB - This prospective, randomized study compared the degree of pulpal anesthesia obtained from an intraosseous injection to an infiltration injection that used 2% lidocaine with 1:100,000 epinephrine. The success rate for the intraosseous injection was 98%; for the infiltration injection, the success rate was 85%. There was no significant difference between the two techniques. The mean time for the onset of pulpal anesthesia was significantly faster with the intraosseous injection and the infiltration injection resulted in a significantly longer duration of pulpal anesthesia. PMID- 15779225 TI - Urgent denture repair in a medically compromised patient. AB - Dental practices will see an increasing number of older patients as our population ages over the next decade. The following article describes a denture repair in a patient with Parkinson's disease. The patient and his caregiver were unable to make multiple trips to the clinic for treatment. The type of repair described in this article minimized the amount of expense and time required to repair the denture. PMID- 15779224 TI - The mineralized and elongated styloid process: a review of current diagnostic criteria and evaluation strategies. AB - The mineralized and elongated styloid process and Eagle's syndrome are similar processes of elongation in which mineralization of the stylohyoid ligament leads to styloid process of the temporal bone. The mineralized and elongated styloid process and Eagle's syndrome differ significantly in terms of the symptoms displayed and the treatment modalities that are sought. The mineralized and elongated styloid process refers to unilateral or bilateral elongation of the styloid process that does not result in any significant pain, discomfort, or limitation of neck movement. It often remains asymptomatic until it is discovered on extraoral radiographs. Eagle's syndrome refers to pain and discomfort in the cervicofacial region resulting specifically from the elongated styloid process. Surgical shortening may be the only treatment that will alleviate the patient's symptoms. This article reviews the entire process of elongation pertaining to the styloid process and discusses the associated syndromes, including current knowledge of the theories of elongation, diagnostic criteria, and treatment strategies. PMID- 15779226 TI - Management of endodontic failures: case selection and treatment modalities. AB - General dentists should realize that endodontic failures can be retreated successfully, avoiding tooth loss. Retreatment of endodontic failure and initial root canal therapy share similar biologic principles and treatment objectives. The criteria for successful root canal therapy are important factors that must be understood before teeth are retreated. This article demonstrates three retreatment modalities for managing endodontic failures as an alternative to tooth extraction. PMID- 15779227 TI - Dental management of liver transplant patients. AB - Dentists are responsible for understanding treatment regimens concerning a myriad of health issues. Increasingly, patients who have significant liver disease or have undergone liver transplant therapy will seek either preoperative or postoperative care. This article reviews current protocols for managing these patients. PMID- 15779228 TI - Identifying causes of dental erosion. AB - Erosion is a chemical process in which an intrinsic or extrinsic source causes a loss of tooth structure. The damage caused by erosion can be accelerated when it occurs in combination with attrition or abrasion. It is important to recognize erosion and determine the source of insult so that destructive behaviors or systemic conditions contributing to the erosive process are terminated. PMID- 15779229 TI - Preventive guidelines for the preschool patient. AB - This article focuses on risk-based caries prevention and specific protective measures for preschool pediatric patients, to provide an age-appropriate approach designed for individual caries risk and optimize preventive care for preschoolers both in-office and at home. PMID- 15779230 TI - Green primary teeth. Diagnosis: Bilirubin deposition. PMID- 15779231 TI - Gene and protein expression profiling of the microvascular compartment in experimental autoimmune encephalomyelitis in C57Bl/6 and SJL mice. AB - Dysfunction of the blood-brain barrier (BBB) is a hallmark of inflammatory diseases of the central nervous system (CNS) such as multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). The molecular mechanisms leading to BBB breakdown are not well understood. In order to find molecules involved in this process, we used oligonucleotide microarrays and proteomics to analyze gene and protein expression of the microvascular compartment isolated from brains of C57Bl/6 and SJL/N mice afflicted with EAE and the microvascular compartment isolated from healthy controls. Out of the 6500 known genes and expressed sequence tags (ESTs) studied, expression of 288 genes was found to be changed. Of these genes 128 were altered in the microvascular compartment in both EAE models. Six proteins were identified to be present at altered levels. In addition to the expected increased expression of genes coding for molecules involved in leukocyte recruitment, genes not yet ascribed to EAE pathogenesis were identified. Thus, proteomics and gene array screens of the microvascular compartment are valid approaches, that can be used to define novel candidate molecules involved in EAE pathogenesis at the level of the BBB. PMID- 15779232 TI - Reduced apoptosis rates in human schwannomas. AB - Schwannomas, tumors originating from Schwann cells, represent a frequent neurological tumor and can occur both in a genetic disorder called neurofibromatosis type 2 (NF2) and sporadically. In both cases the genetic background is identical as all schwannomas are caused by biallelic mutations in the tumor suppressor gene NF2 coding for merlin. Mutations in this gene have also been found to be responsible for 50% to 60% of spontaneous and 100% of the NF2 associated meningiomas. The NF2 gene product, merlin, links transmembrane proteins to the cytoskeleton and is involved in intracellular signaling processes. It has previously been shown that reexpression of wild-type merlin in primary human schwannoma cells leads to an increase in the number of apoptotic cells. Here, we report in vivo and in vitro evidence that the basal apoptosis rate of primary human schwannoma cells is reduced in comparison to that of normal Schwann cells, supporting the idea that in this benign tumor type, apoptosis has a role in tumorigenesis. PMID- 15779233 TI - Chromosome 22q deletions in atypical teratoid/rhabdoid tumors in adults. AB - Atypical teratoid/rhabdoid tumors (AT/RTs) are rare, malignant brain tumors that usually occur in the posterior fossa. Both AT/RT and the analogous tumor outside the brain, malignant rhabdoid tumor, share a polyphenotypic immunoprofile and frequent 22q deletions with inactivation of the IN11/hSNF5 gene. Reports, so far, indicate that AT/RTs occur almost exclusively in children, most of whom are 5 years-old or less. The rarity of the tumor and the polyphenotypic immunoprofile, characterized by antigen expression that is often patchy, make diagnosis in adults difficult and controversial. We describe three AT/RTs in adults in which the diagnoses were supported by detection of 22q11.2 deletions, INI1 mutation and/or loss of INI1 protein expression. Two patients were female, ages 20 and 31 and one was male, age 45. Two tumors occurred in the sella or sellar region and one in the cerebellum. In all cases, fluorescence in situ hybridization with probes to the BCR (22q11.2) and NF2 (22q12) regions of chromosome 22 revealed single copy deletions of BCR with normal dosages of NF2 and, in all cases, immunohistochemistry demonstrated loss of INI1 protein expression. In one case, a single base pair deletion was detected in the INI1/hSNF5 gene. These molecular findings confirm the occurrence of AT/RTs in adults. Although rare, AT/RT should be considered in the differential diagnosis of poorly differentiated intracranial tumors in adults. PMID- 15779234 TI - Profound cardiac sympathetic denervation occurs in Parkinson disease. AB - In the last few years, cardiac sympathetic dysfunction in Parkinson disease (PD) has been postulated on the basis of decreased cardiac uptake of sympathoneural imaging tracers. However, the pathological substrate for the dysfunction remains to be established. We examined the left ventricular anterior wall from postmortem specimens with immunohistochemical staining for tyrosine hydroxylase (TH), neurofilament (NF) and S-100 protein in PD patients and control subjects, and quantified the immunoreactive areas. As TH-immunoreactive axons nearly disappeared and NF-immunoreactive axons drastically decreased in number, the morphological degeneration of the cardiac sympathetic nerves in PD was confirmed. Quantitative analysis showed that sympathetic nerves were preferentially involved. Triple immunofluorolabeling for NF, TH, and myelin basic protein showed clearly the profound involvement of sympathetic axons in PD. The extent of involvement of the cardiac sympathetic nerves seems likely to be equivalent to that in the central nervous system, including the nigrostriatal dopaminergic system. PD affects the cardiac sympathetic nervous system profoundly as well as nigrostriatal dopaminergic system. PMID- 15779235 TI - Ephrin A receptors and ligands in lesions and normal-appearing white matter in multiple sclerosis. AB - Complexes of the tyrosine kinase ephrin ligands (ephrins) and their receptors (Ephs) provide critical cell recognition signals in CNS development. Complementary ephrin/Eph expression gradients present topographic guidance cues that may either stimulate or repulse axon growth. Some ephrin/Ephs are upregulated in adult CNS injury models. To assess their involvement in multiple sclerosis (MS), ephrin A1-5 and Eph A1-8 expression was analyzed in CNS tissues using immunohistochemistry. Control samples showed distinct expression patterns for each ephrin/Eph on different cell types. Perivascular mononuclear inflammatory cells, reactive astrocytes and macrophages expressed ephrin A1-4, Eph A1, -A3, -A4, -A6 and -A7 in active MS lesions. Axonal ephrin A1 and Eph A3, A4, and -A7 expression was increased in active lesions and was greater in normal appearing white matter (NAWM) adjacent to active lesions than within or adjacent to chronic MS lesions, in contralateral NAWM, or in control samples. As in development, therefore, there are temporally dynamic, lesion-associated axonal ephrin/Eph A expression gradients in the CNS of MS patients. These results indicate that ephrin/Eph As are useful cell markers in human CNS tissue samples; they likely are involved in the immunopathogenesis of active lesions and in neurodegeneration in MS NAWM; and they represent potential therapeutic targets in MS. PMID- 15779236 TI - Genes associated with fast glioma cell migration in vitro and in vivo. AB - Identification of genes mediating glioma invasion promotes the understanding of glia motility and might result in biologically based therapeutic approaches. Most experimental studies have been performed in vitro, although glial cells typically undergo marked phenotypic change following placement into cell culture. To evaluate migration mechanisms operating in vitro versus in vivo, we used C6 rat glioblastoma cells for selecting highly migratory cells in a monolayer migration assay as well as in brains of nude mice, and analyzed in each paradigm the expression profiles of these "fast" cells versus those of the original "slow" cells using oligonucleotide microarrays comprising 8832 genes. In vitro, 516 (10.6%) of 4848 expressed genes were regulated (i.e., differentially expressed in fast versus slow cells); 916 genes were expressed only in vitro, including 142 (15.5%) regulated genes. In vivo, 245 (6.1%) of 4044 expressed genes were regulated; 112 genes were expressed only in vivo, including 25 (22.3%) regulated genes, none of them having a known relation to glioma invasion. Of 730 regulated genes, only 31 (4.2%) were regulated in parallel in vitro and in vivo, most of them having a known relation to (glioma) invasion. Our data provide new molecular entry points for identifying glioma invasion genes operating exclusively in the brain. They further suggest that genes underlying glia cell motility are strikingly different in vitro and in vivo. PMID- 15779237 TI - Insights into meningioangiomatosis with and without meningioma: a clinicopathologic and genetic series of 24 cases with review of the literature. AB - Meningioangiomatosis (MA) is a rare seizure-associated lesion of presumed hamartomatous or developmental origin. It is occasionally combined with a neoplasm, most commonly meningioma (MA-M). In the current study, we examined 24 cases (14 pure MA, 10 MA-M) using immunohistochemistry for merlin, protein 4.1 B, progesterone receptor (PR), and MIB-1, as well as FISH for NF2 and 4.1B gene dosages. Nine cases of MA-M (90%) had gene deletions (NF2/4.7B), protein losses (merlin/protein 4.1B), and/or PR positivity, with a similar or identical phenotype in both components. No PR positivity or gene deletions were seen in pure MAs, though merlin and/or protein 4.1B were immunonegative in six cases. Our data suggest that in most MA-Ms, the MA component is neoplastic, likely representing an exuberant perivascular pattern of spread from the meningioma, rather than an underlying hamartoma. This pattern of spread may be facilitated by meningiomas that are predominantly leptomeningeal or intracerebral in origin. It remains important to distinguish this pattern from true brain invasion, given the more ominous prognostic significance of the latter. In contrast, most perivascular spindled cells of pure MA are genetically and immunohistochemically similar to non-neoplastic meningothelial cells, consistent with current histogenetic theories. PMID- 15779238 TI - Intracellular and extracellular Abeta, a tale of two neuropathologies. AB - The central pathological cause of Alzheimer disease (AD) is hypothesized to be an excess of beta-amyloid (Abeta) which accumulates into toxic fibrillar deposits within extracellular areas of the brain. These deposits disrupt neural and synaptic function and ultimately lead to neuronal degeneration and dementia. In addition to the pathological roles attributed to Abeta, evidence from our laboratory would suggest that Abeta serves a physiological role in the modulation of CRE-directed gene expression. This commentary also highlights some of the pathological consequences of the accumulation of intracellular Abeta. Finally it discusses the impact of cortical Abeta burden on transmitter-specific synaptic numbers as well as the generation of dystrophic neurites. The fundamental thesis of my proposal is that the Abeta pathology seen in AD is a continuous process from an initial abnormal Abeta intracellular accumulation to the well-established extracellular Abeta aggregation, culminating in the formation of amyloid plaques and dystrophic neurites. PMID- 15779239 TI - Immunological and anti-chaperone therapeutic approaches for Alzheimer disease. AB - Alzheimer disease (AD) is the most common cause of dementia. Currently available therapies only provide symptomatic relief. A number of therapeutic approaches are under development that aim to increase the clearance of brain Abeta peptides. These include immune mediated clearance of Abeta and the inhibition of the interaction between Abeta and its pathological chaperones. Both active and passive immunization has been shown to have robust effects in transgenic mouse models of AD on amyloid reduction and behavioral improvements. However, a human trial of active immunization has been associated with significant toxicity in a minority of patients. New generation vaccines are being developed which likely will reduce the potential for cell-mediated toxicity. In addition, the recent development of anti-chaperone therapy opens a new therapeutic avenue which is unlikely to be associated with toxicity. PMID- 15779240 TI - Neurovascular pathways and Alzheimer amyloid beta-peptide. AB - According to the prevailing amyloid cascade hypothesis, the onset and progression of a chronic neurodegenerative condition in Alzheimer disease (AD) is initiated by the amyloid beta-peptide (Abeta) accumulation in brain and consequent neuronal toxicity. Recent emphasis on co-morbidity of AD and cerebrovascular disease and the recognition that cerebrovascular dysregulation is an important feature of AD, has shed new light on neurovascular dysfunction as a possible contributor to cognitive decline and Alzheimer neurodegeneration. In the same time, this association has raised a question as to whether there is a causal relationship between cerebrovascular dysregulation and Abeta-initiated pathology, and whether influencing targets in the neurovasculature may prevent different forms of Abeta brain accumulation and/or lower pre-existing accumulates in a later stage of the disease. Pathogenic cascades which operate to dissociate normal transport exchanges between central and peripheral pools of Abeta, and decreased vascular competence leading to brain hypoperfusion and impaired Abeta clearance are discussed. We suggest that there is a link between neurovascular dysfunction and elevated brain Abeta which provides a new scenario for therapeutic interventions to control Alzheimer mental deterioration. PMID- 15779242 TI - Re: Interferon-gamma expression in periventricular leukomalacia in the human brain. PMID- 15779241 TI - The Abeta hypothesis: leading us to rationally-designed therapeutic strategies for the treatment or prevention of Alzheimer disease. AB - In recent years the amyloid cascade hypothesis of Alzheimer disease (AD) has been increasingly referred to as the amyloid beta protein (Abeta) cascade hypothesis. This subtle rephrasing reflects the acknowledgment that there is debate within the field as to whether Abeta aggregates other than Abeta deposited as classic amyloid fibrils could trigger the pathological cascade that results in neuronal dysfunction and neurodegeneration. Despite this semantic shift, which highlights one enigmatic aspects of AD, the evidence supporting the Abeta hypothesis of AD is extensive. More importantly the Abeta hypothesis of AD has led and will continue to lead to the development of rationale therapeutic strategies that are likely to either prevent or treat this devastating disease. In this review, the evidence supporting the Abeta hypothesis and the recent advances in anti-Abeta therapy are discussed. PMID- 15779243 TI - July 2004: 40-year-old man with headaches and dyspnea. AB - A 40-year-old man had a 6-week history of severe frontal headaches and dry cough. Chest x-ray showed hilar adenopathy with bilateral parenchymal infiltrates. A diagnosis of atypical pneumonia was made. Four weeks later he was admitted with persistent headache. Infectious screen was negative. Brain MR post contrast, revealed cerebellar enhancement and swelling with moderate tonsillar herniation; findings which precluded the performance of a lumbar puncture. High resolution CT thorax confirmed hilar abnormalities; shown by microscopy to represent non caseating granulomata. A presumptive diagnosis of sarcoidosis was reached. Despite an initial symptomatic improvement his headache persisted. Repeat MRI, eleven days after admission, showed reduced cerebellar enhancement and swelling with no change in the degree of tonsillar herniation. He deteriorated acutely and died two weeks after admission. Autopsy revealed cerebral oedema with tonsillar herniation secondary to cryptococcal meningitis variety neoformans. There was no evidence of neurosarcoid. Active and inactive sarcoid was identified in the lungs and hilar nodes with no evidence of systemic sarcoid. Focal evidence of cryptococcal pneumonitis was present in the lung as a necrotic focus. A strong index of clinical suspicion is necessary to diagnose the rare association of cryptococcus complicating sarcoidosis. PMID- 15779244 TI - August 2004: 64-year-old man with intermittent paresthesia of the abdomen and of the legs. AB - A 64-year-old-man had a 2-year history beginning with a sense of abdominal "constriction." Additional slowly rising symptoms, such as tingling of the legs, mild gait ataxia and painful micturition, led to MRI investigation of the spinal cord. A fusiform enlargement of the cord extending from T5 to T8 was shown. The space occupying lesion infiltrated diffusely the spinal cord. A contrast medium enhancing exophytic tumor pellet was approached via a 2-level laminoplasty and resected. Biopsies were taken from different exophytic tumor areas whereas the intramedullary part was spared. The histologic examination confirmed the typical pattern of a pilocytic astrocytoma in all specimens. In our surgical experience with 226 intramedullary tumors and with 117 patients affected by intracranial pilocytic astrocytoma this case is unique because of its combination of tumor location, growth pattern and age of the patient. PMID- 15779245 TI - September 2004: a 6-year-old girl with headache and stiff neck. AB - Free-living amebas in the genera Naegleria, Acanthamoeba and Balamuthia are known to cause CNS infections. Here we report a case of fatal granulomatous amebic meningoencephalitis (GAE) caused by Balamuthia mandrillaris in a 6-year-old previously healthy girl who presented with headache and stiff neck. She was treated medically for brain abscess after a CT scan identified a ring-enhancing lesion in the right temporo-parietal area. A brain biopsy showed necrosis and granulomatous inflammation. Subsequently, multiple new lesions appeared in the brain bilaterally. A second brain biopsy revealed viable amebic trophozoites that were most abundant in perivascular spaces, accompanied by neutrophils, macrophages and eosinophils. Immunofluorescence study confirmed the amoeba as Balamuthia mandrillaris. This case demonstrates that making diagnosis of GAE pre mortem requires a high index of suspicion. Amebic infection should be included in the differential diagnosis of any granulomatous lesion in CNS; and careful search for amebic parasites should be carried out especially when necrosis predominates in the pathological material. PMID- 15779246 TI - Can diabetes educators become "contagious?". PMID- 15779247 TI - Implementing an empowerment-based diabetes self-management education program. AB - Diabetes educators are challenged to develop culturally appropriate, integrated, behaviorally based, effective education programs. This article describes the intervention used in a problem-based educational program for urban African Americans with diabetes. The intervention consisted of six 2-hour, weekly group educational and data collection sessions. No lectures were used, and the content was determined by participants' questions and concerns. Culturally tailored written educational materials were also provided. PMID- 15779248 TI - Diabetes foot self-care practices in a rural triethnic population. AB - PURPOSE: The purposes of this study were to assess the level of foot self-care performed in a rural, multiethnic population of older adults and to identify factors associated with foot self-care. METHODS: The Evaluating Long-term Diabetes Self-management Among Elder Rural Adults study included a random sample of 701 African American, Native American, and white adults from 2 rural North Carolina counties. Participants completed in-home interviews, 5 foot self-care practices from the Summary of Diabetes Self-Care Activities (SDSCA), functional status measures, and measures of education and support for foot care. RESULTS: Foot care practices/behaviors reported at least 6 days/week ranged from 35.6% for inspecting shoes to 79.2% for not soaking feet. Four independent predictors of the SDSCA summary foot care index score were observed: having been shown how to care for feet (P < .0001), female gender (P = .03), having had a doctor check nerves in feet in past year (P = .02), and not receiving support caring for feet (P = .0425). CONCLUSIONS: These findings indicate that educating patients about foot self-care may encourage routine foot care but that those dependent on either formal or informal support to perform foot care do so less frequently than those who perform it independently. PMID- 15779249 TI - Self-monitoring of blood glucose in a multiethnic population of rural older adults with diabetes. AB - PURPOSE: The purpose of the study was to describe self-monitoring of blood glucose (SMBG) practices of 698 older adults with type 2 diabetes in the rural Southeast, to identify characteristics differentiating testers from nontesters, and to identify personal and support-related predictors of monitoring frequency. METHODS: The ELDER (Evaluating Long-term Diabetes Self-management Among Elderly Rural Adults) study was a population-based, cross-sectional survey of African American, Native American, and white Medicare recipients > or =65 years with diagnosed diabetes. Data were obtained through in-home interviews. Multiple logistic regression models were used to identify factors associated with SMBG and frequency of monitoring. RESULTS: Seventy-seven percent of respondents practiced SMBG in the previous week; 40% tested every day in that week. No ethnic differences were seen. Significant independent predictors of any SMBG were medication regimen (taking oral agents or insulin with or without oral agents) and health care provider (HCP) recommendation to test. Among those monitoring, significant independent predictors of SMBG frequency were medication regimen, HCP recommendation to test, duration of diabetes, and receiving help with testing, which was negatively associated with monitoring frequency. CONCLUSIONS: Among rural older persons with diabetes, HCP recommendation significantly affected practicing SMBG and SMBG frequency. These findings suggest points of intervention by diabetes educators with this vulnerable population. Further research is needed to determine how older adults use SMBG data in their self-care regimen. PMID- 15779250 TI - Developing a new generation of ongoing: Diabetes self-management support interventions: a preliminary report. AB - PURPOSE: The study examined the feasibility, acceptability, and potential impact of an innovative, community-based, ongoing self-management intervention aimed at enhancing and sustaining self-care behaviors over the long term among urban African Americans with type 2 diabetes. METHODS: Sixty-two African American men and women completed the study. Participants were invited to attend 24 weekly, consecutive, diabetes self-management support/ education groups. The flow of the weekly group sessions was guided by questions and concerns of the patients. Baseline and 6-month follow-up metabolic functioning, lipid profiles, cardiovascular functioning, and self-care behaviors were assessed. RESULTS: Ninety percent (n = 56) of the sample attended at least 1 session; 40% attended at least 12 or more sessions. Paired t tests found significant improvements in body mass index (P < .001), total cholesterol (P < .01), high-density lipoprotein (P < .05), and low-density lipoprotein (P < .001). Significant increases were also found for self-care behaviors (P < .05). CONCLUSIONS: Preliminary evidence suggests that participation in this weekly problem-based, self-management support intervention can yield diabetes-related health benefits. PMID- 15779252 TI - Understanding skin care. PMID- 15779251 TI - "Plenty of sickness": descriptions by African Americans living in rural areas with type 2 diabetes. AB - PURPOSE: The purposes of this study were to (1) describe facilitators and barriers to self-care for African Americans with type 2 diabetes living in a rural community, (2) compare experiences of men and women, and (3) solicit recommendations for programs of care. METHODS: Focus groups with small numbers of men and women (n = 7) were held to explore facilitators and barriers to self management. Groups were led by moderators of the same gender and race as participants. Data from these information-rich cases were analyzed thematically using FolioViews and Inspiration software. RESULTS: Final categories included "being diagnosed," "diabetes as betrayal by the body," "provider-individual family relationship," "self-management," and "difficulty getting help." Although limited by sample size and methodology, results indicate differences by gender that require additional investigation. CONCLUSIONS: Knowledge of self-care patterns is foundational to designing culturally appropriate interventions and programs of care for rural African Americans living with diabetes. PMID- 15779253 TI - [Increased survival following antibiotic prophylaxis in patients with cirrhosis and upper gastrointestinal hemorrhage]. PMID- 15779254 TI - [Antibiotic prophylaxis for patients with liver cirrhosis and upper gastrointestinal hemorrhage. Is there evidence for its use?]. PMID- 15779255 TI - [Liver cirrhosis and bacterial infections]. PMID- 15779256 TI - [Antibiotic prophylaxis in variceal bleeding to patients with liver cirrhosis? Yes!]. PMID- 15779257 TI - [The molecular-genetic background of spinal muscular atrophy. Diagnosis, prognosis and future treatment]. PMID- 15779258 TI - [Chemotherapy as a supplement to radical treatment of patients with locally advanced bladder cancer]. PMID- 15779259 TI - [Legal abortion. Why should it be well taken care of?]. PMID- 15779260 TI - [Knot tying techniques]. PMID- 15779261 TI - [Incidence of tetanus in Denmark, 1983-2000]. PMID- 15779262 TI - [The societal cost of zoonotic Salmonella infections and other foodborne bacterial infections in Denmark]. PMID- 15779263 TI - [Prolonged restaurant-associated outbreak of multidrug resistant Salmonella typhimurium--secondary publication]. PMID- 15779264 TI - [Genetically determined reduction in antioxidative protection and increased risk of ischemic heart disease--secondary publication]. PMID- 15779265 TI - [An outbreak of methicillin-resistant Staphylococcus aureus infection among patients with liver cirrhosis]. PMID- 15779266 TI - [Tularemia acquired in Denmark by an eight-year-old child]. PMID- 15779267 TI - [Second-look laparoscopy in intestinal ischemia]. PMID- 15779268 TI - [Questions to the Drug Committee about administration of Lyrica]. PMID- 15779269 TI - [Mammography screening--again]. PMID- 15779270 TI - [Magnyl 75 mg daily as prevention against migraine?]. PMID- 15779271 TI - [Chronic fatigue syndrome, an acknowledged neurologic diagnosis]. PMID- 15779272 TI - [Ezetimibe and safety]. PMID- 15779273 TI - Electronic health records: taking the plunge. PMID- 15779274 TI - E-H-R spells trouble. PMID- 15779275 TI - More EHR woes. PMID- 15779276 TI - How will data standards affect EHRs? PMID- 15779277 TI - Full financial disclosure would help EHR assessment. PMID- 15779278 TI - New year, new Medicare benefits. PMID- 15779279 TI - Answers to your questions. PMID- 15779280 TI - Purchasing an affordable electronic health record. PMID- 15779281 TI - Practicing without paper charts: one clinic's experience. PMID- 15779282 TI - Electonic health records: a user-satisfaction survey. PMID- 15779283 TI - How to select an electronic health record system. PMID- 15779285 TI - Headache. PMID- 15779284 TI - Managing the habitual no-show patient. PMID- 15779286 TI - Making the most of every minute. PMID- 15779287 TI - [New options in pharmacological treatment of urge urinary incontinence in women]. AB - OBJECTIVE: The aim of this paper is to provide a brief update review of current pharmacological agents used to treat women with urge incontinence. DESIGN: Review article. SETTINGS: Gynecological and Obstetric Clinic, 1. LF UK and VFN, Prague. RESULTS: Urge urinary incontinence is the condition characterized by the involuntary loss of urine accompained by a strong desire to void. Urge incontinence is often due to detrusor instability, although the instability of the detrusor bladder muscle cannot be demonstrated clinically but through cystomery. Sensory urge incontinence is the involuntary loss of urine associated with urgency and a strong desire to void urine immediately due to the hypersensitivity of the bladder and urethral sensory receptors. The mainstay treatment for this common form of incontinence is conservative treatment. There are two forms of conservative treatment: behavioural techniques and pharmacotherapy. The urge urinary incontinence is most successfully treated by a drug therapy. Anticholinergic drugs and anticholinergic antispasmodic drugs are the primary pharmacologic treatment for this condition, although the usefulness of this agent has been limited by a lack of selectivity for the bladder, which gives rise to frequent, bothersome side effect (dry mouth, constipation, blurred vision, etc.) For these reasons, tolterodine was developed as the first antimuscarinic agent specifically targeted for the treatment of the urge urinary incontinence. This agent has demonstrated a bladder-selective profile in vivo, leading to a more pronunced and longer lasting effect on the bladder than on salivation in humans. PMID- 15779288 TI - [Importance of registry of aggregated data on women giving birth]. AB - OBJECTIVE: Analysis of causes of unchanging level of national perinatal mortality and identification of potential for its further decrease. DESIGN: Retrospective epidemiological analysis of aggregated data on perinatal care. SETTING: Mother and Child Care Institute, Prague 4-Podoli. METHODS: Correlation of selected national aggregated data on the level of perinatal care under current system and under influence of changing conditions for care provision. RESULTS: The analysis of some criteria of perinatal care demonstrated the influence of several contradictory processes: 1) among newborns below 2500 grams the stillbirth increased while the early neonatal mortality decreased; there was also decrease of contribution of congenital malformation in this group, 2) although there were more live born babies under 500 grams, who suffered from high mortality, their contribution to perinatal mortality was compensated by further decrease of early neonatal mortality of newborns weighing 1000-1499 grams, 3) the increasing frequency of low birthweight newborns did not influence the total perinatal mortality due to improved perinatal care. We observed the improvement of results among half of the regional perinatal centres with formerly higher early neonatal mortality figures, which represents the decrease of one of the reserves for further decrease of early neonatal mortality. There is a similar situation in centralisation of preterm deliveries--transfer in utero--which reached its maximum. Up to now we did not succeed in early referral of women with serious pregnancy complications which lead to stillbirth to perinatal centres with lower stillbirth rates. However this represents only small potential for decrease of total perinatal mortality. CONCLUSION: The 4-year stagnation of national perinatal mortality is a result of contradictory influence of improving level of care on one side and worsening of conditions on the other side. The potential for improvement under current conditions is almost exhausted. PMID- 15779289 TI - [Analysis of maternal mortality in the Czech Republic in 2001]. AB - OBJECTIVE: Analysis of maternal mortality in the Czech Republic in 2001. DESIGN: Retrospective statistical and clinical analysis. SETTING: Mother and Child Care Institute, Prague-Podoli. METHODS: We used database of 9 cases of maternal deaths in the Czech Republic during 2001 to analyze their causes, clinical courses especially related to obstetrical surgery, adequacy of provided care, and characteristics of their occurrence. RESULTS: There were total of 9 reported maternal deaths in pregnancy, during labor or within 42 days after delivery in the Czech Republic in 2001. During the same period there were 90,715 live births in the Czech Republic, and Pregnancy-related mortality ratio (A+B+C) was 0.0992/1000, i.e. 9.92 deaths per 100,000 live births. This is about 1.08/1000 better than in 2000 (the table 5 demonstrates the development of maternal mortality during the last 11 years). One reported death was unrelated to gestation (category C), therefore adjusted maternal mortality rate (A+B) was 0.0882/1000, i.e. 8.82 deaths per 100,000 live births versus 9.9 deaths per 100,000 live births in 2000. Group A (specific risk-direct maternal mortality) contributed to adjusted maternal mortality by 5 maternal deaths and direct maternal mortality was 5.5 per 100,000 live births. Group B (non-specific risk undirect maternal mortality) contributed by 3 maternal deaths and undirect maternal mortality was 3.3 per 100,000 live births. PMID- 15779290 TI - [Analysis of present diagnostic methods of intrapartum fetal hypoxia]. AB - OBJECTIVE: To evaluate specificity of present diagnostic methods of intrapartal fetal hypoxia (cardiotocography--CTG, fetal pulse oxymetry--FpO2, ST-ECG analysis). DESIGN: A prospective study. SETTING: Gynecology-Obstetrics Clinic, 1st Medical Faculty and General Faculty Hospital in Prague. METHODS: The results of synchronic fetus monitoring by means of CTG, FpO2 and STAN-ST 21 were followed in 53 mothers with a risk and pathological delivering of birth in the period of April 2003 to March 2004. The study investigated, which of the methods provided the best prediction of the intrapartal fetal hypoxia. The statistical evaluation (p-mark test and Mc Namara test) was based on the assessment of correct or incorrect prediction of the Apgar score values in the first minute after birth, pH in umbilical artery and the lactate level. We also investigated results of three methods during postpartum depression, turbid or mushy amniotic fluid and the way the individual methods were made useful in indication for ending the delivery. RESULTS: In comparison with CTG there was a statistically significantly higher specificity in FpO2 and STAN in the evaluation of Apgar score in the newborn in the first minute after birth, FpO2 (p=0.007) and STAN-ST (p<0.001), in the determination of pH (a) from umbilical blood FpO2 (p=0.029) and STAN (p=0.001) and the occurrence of postpartum depression of the newborn in minute 30 60 after birth FpO2 (p=0.019) and STAN (p=0.0005). The changes in lactate level in umbilical blood were better predicated by STAN (p=0.001). FpO2 evaluated the changes in the same way as CTG. The threatening hypoxia in strongly turbid or even mushy amniotic fluid was correctly evaluated by STAN only (p=0.002). The FpO2 evaluation was not statistically significant. There was not any statistically significant difference in the indication of operation for ending the delivery among the individual methods. CONCLUSIONS: The results univocally demonstrated that the used of other method for diagnosis on intrapartal fetus hypoxia--Fetal pulse oxymetry and ST--analysis of ECG of the fetus give more precision to the diagnosis. The introduction of these methods requires a correct interpretation and the effort of the obstetrician to use these methods in clinical practice. PMID- 15779291 TI - [Contribution of Doppler examination in pregnancy at risk of alloimune fetus anemia]. AB - OBJECTIVE: To evaluate the utilisation measurements of peak systolic velocities in the middle cerebral artery (MCA-PSV) to predict fetal anemia in pregnancies complicated by alloimmune antibodies known to cause immunological hydrops. DESIGN: A prospective study. SETTING: Department of Obstetrics and Gynecology, University Hospital, Olomouc. METHODS: In 38 pregnancies at risk for fetal anemia due to maternal red-cell alloimmunization MCA-PSV had been assessed and fetal blood sampling for measurement of hemoglobin concentration was obtained subsequently, either by cordocentesis or at delivery. RESULTS: 66 examinations were performed at 19-37 week's gestation. An MCA-PSV >1.5 MoM detected 100% of severely anemic fetuses with a hemoglobin concentration <0.65 MoM that required invasive intervention. In 18 cases a cordocentesis was performed and intrauterine blood transfusion was given alternatively. Remaining fetuses did not require invasive intrauterine intervention and no or mild hemolytic anemia and hyperbilirubinemia were diagnosed after delivery. No false positive case was identified (enregistered). The median maternal age at the term of delivery was 29.1 (range, 19-41) years. CONCLUSION: Middle cerebral artery peak systolic velocity is a highly sensitive non-invasive means for determining the degree of anemia. A Doppler interval of seven days is recommended. PMID- 15779293 TI - [Birth defects in the Czech Republic--expected incidence in the coming years]. AB - OBJECTIVE: The study was aimed to analyze expected incidence of birth defects as a whole and some selected types of birth defects on the basis of the mean values over the period of 1983-2002 in the Czech Republic. A possible incidence of birth defects in the Czech Republic in the coming years was evaluated. DESIGN: A statistical estimate and analysis of the expected incidence based on retrospective values of incidence of birth defects from the data of the Institute of the Care of Mother and Child (UPMD) and the Institute of Medical Information and Statistics of the Czech Republic (UZIS). METHODS: The estimate of incidence of birth defects as a whole as well as according to individual selected types with the application of exponential adjustment of time series with the time weighted parameter for individual years of the retrospective period of time. RESULTS AND CONCLUSIONS: The statistical estimate indicates that 1,244,767 children should be born in the Czech Republic over the period of 2003-2015. In this period of time there should be 50,706-67,628 prenatally and postnatally diagnosed cases of birth defects in this republic. These our results represent an estimate of birth defects in the Czech Republic in the coming years and draw attention to the importance of registry of birth defects in the Czech Republic. The epidemiology of birth defects actually represents a multidisciplinary branch, which is primarily aimed at and providing data for the evaluation of quality of prenatal and postnatal care in the Czech Republic. PMID- 15779292 TI - [Risk factors for 3rd and 4th degree perineal ruptures during delivery]. AB - OBJECTIVE: Summary of the mutual relationship between vaginal delivery and anal incontinence. DESIGN: Retrospective case-control study. SETTING: Department of Gynaecology and Obstetrics, Charles University and Faculty Hospital Pilsen, Czech Republic. METHODS: 9600 vaginal deliveries between January 1997 and March 2004 were evaluated. The aim of the study was to observe the incidence of the 3rd degree tear and identification of the risk factors. The recognition of the perineal injury was open to the individual interpretation and assessment of the doctors involved. RESULTS: The incidence of the 3rd degree tear was 0.84%. The diagnostics of the 3rd degree tear was significantly more frequent if the evaluation of the perineal injury was performed by the urogynaecologists (p<0.001). Statistically significant risk factors of the 3rd degree tear in the univariate and multivariate analysis were: forceps, shoulder dystocia, the length of the 2nd stage of labour and maternal cooperation at the final phase of delivery. Parturity (primiparity), birth weight and episiotomy (when all types of episiotomy were included) were statistically significant only in the univariate analysis. Mediolateral episiotomy was not proved to be either the risk or the protective significant factor. SUMMARY: The diagnostics of the 3rd degree tear demands a detailed knowledge of the structure and anatomy of the pelvic floor. A sufficient amount of experience and erudition are the requirements for the prevention, diagnostics and subsequent treatment of this injury. PMID- 15779294 TI - [Pelvic exenteration]. AB - OBJECTIVE: Review of literature concerning pelvic exenterative procedures. DESIGN: Review article. SETTING: Department of Obstetrics and Gynecology, Charles University, Prague. METHODS: Review and critical assessment of published data. CONCLUSION: Pelvic exenterations are standard procedures in oncogynecology which have no alternative in certain indications. The most frequent indications are recurrences or progressions of cervical, vulva or vaginal cancers. Exenterative procedures might be used in primary treatment in some cases of locally advanced tumors. Mortality of current procedures reaches 5% to 10%, early and late postoperative morbidity is frequent (40-60%). Recently explorative laparoscopy is used in preoperative staging to decrease the number of aborted procedures due to distant metastasis or pelvic tumor inoperability. The procedure with high morbidity, causing impairment of quality of life, is justified due to good follow up results--5-years overall survival is about 50-60%. It should be emphasized that with no treatment in these patients median of survival reaches about 6 months. PMID- 15779295 TI - [Possibilities of increased radicality in pelvic exenteration]. AB - OBJECTIVE: Review of advanced radicality in pelvic exenterations. DESIGN: Review article. SETTING: Department of Obstetrics and Gynecology, General Faculty Hospital and Ist Medical Faculty of the Charles University, Prague. METHODS: Review and critical assessment of published data. CONCLUSIONS: More extensive radicality in pelvic exenterations make possible to use surgical treatment in some cases of lateral recurrences or central recurrences with the attachment to the pelvic side wall. One possible technique is a combination of en bloc exenteration with pelvic bone resection, most frequently sacrum. Experiences in gynecological tumours are so far limited. Laterally extended resection was described in lateral infrailiac pelvic wall recurrences. The only one presented paper related to laterally extended procedures showed a reasonable overall survival of patients; however follow-up is limited. PMID- 15779296 TI - [Abdominal hysterectomy--risk factor in development of urinary incontinence? Results of a questionnaire study]. AB - OBJECTIVE: To evaluate the risk of urinary incontinence development after abdominal hysterectomy for benign gynecological disease. DESIGN: Retrospective questionnaire study. SETTING: Obstetrics and Gynecology Department, Charles University 2nd Medical Faculty and Teaching Hospital Motol, Prague. METHODS: Questionnaire study of women in which abdominal hysterectomy for benign gynecological disease was performed from January till December 2001 at our Department of Gynecology and Obstetrics. The questionnaire included questions about symptoms of stress, urge and mixed urinary incontinence. They were evaluated in connection with age, parity, hormonal status and body mass index. Student t-test was used for the statistical analysis. RESULTS: 114 (59.7%) from all sent-out questionnaires were obtained. In 15.8% cases (18 from 114) de novo urinary incontinence after surgery developed. 77.8% (14 from 18) from these patients suffered from genuine stress incontinence. The rest (4 from 18) of the women had symptoms of mixed incontinence. Study results draw attention to 13.1% women (15 from 114) in which preoperative stress incontinence persisted even after hysterectomy. Preoperative stress incontinence was cured in 6.1% (7 from 114) cases after hysterectomy. Age, parity, hormonal status and body mass index were not statistically different in all groups after surgery. CONCLUSIONS: The results of the study show relatively high risk of urinary incontinence development after abdominal hysterectomy in previously continent women. The results also show high persistence of the stress incontinence symptoms in women who did not inform about their incontinence even if they were asked about it. Higher quality of further health education of the women with drawing their attention to both the possibility of surgical treatment of urinary incontinence simultaneously with hysterectomy are the most important aspects how to make the current gynecological surgical therapy more effective. PMID- 15779297 TI - [MMP-26 mRNA and estrogen receptor alpha co-expression in normal and pathological endometrium]. AB - OBJECTIVE: To examine the expression pattern of matrix metalloproteinase-26 (MMP 26) mRNA and estrogen receptor-alpha (ER alpha) in normal, hyperplastic, premalignant and malignant endometrial tissue. DESIGN: Experimental study. SETTING: Department of Obstetrics and Gynecology of the Palacky University Medical School and University Hospital, Olomouc, Czech Republic, Department of Obstetrics and Gynecology, University Hospital, Lund, Sweden, Atherosclerosis Research Unit, King Gustav V Research Institute, Karolinska Hospital, Stockholm, Sweden. METHODS: We studied MMP-26 mRNA and ER alpha in 36 normal, 7 hyperplastic, 6 premalignant and 19 malignant endometrial samples. Based on histological examination, all normal specimens were classified according to an ideal 28 day menstrual cycle as early, mid, and late proliferative phase, early, mid and late secretory phase and menstrual phase. Samples with hyperplasia were classified as simple or complex. Premalignant samples were represented by complex hyperplasia with atypia. Malignant samples were histologically classified as well, intermediately and poorly differentiated, respectively. Specimens were analyzed using in situ hybridization and real time PCR. ER alpha was localized by immunohistochemistry. RESULTS: Epithelial MMP-26 mRNA expression was highest in the early secretory phase and in endometrial hyperplasia. Expression levels were low in the late secretory and menstrual phase and in malignant samples decreased gradually with dedifferentiation. Expression pattern of MMP-26 mRNA in normal, hyperplastic, premalignant and malignant endometrial tissue strongly co-variated with that of ER alpha. CONCLUSION: Co-expression of MMP-26 and ER alpha in normal and pathological endometrial tissue suggests possible regulation of MMP-26 gene by estrogen. PMID- 15779298 TI - [Protein bcl-2 expression in the secretory endometrium after oral estrogen progesterone replacement]. AB - OBJECTIVE: To compare the changes of the protein bcl-2 expression, regulatory mechanism in the process of apoptosis, in the secretory endometrium throughout natural and artificial (estrogen-progesterone substitution) cycles in the same group of patients. DESIGN: A prospective study. SETTING: Dept. of Obstetrics and Gynecology, University Hospital Olomouc, Dept. of Histology and Embryology, Faculty of Medicine, Palacky University in Olomouc, Dept. of Obstetrics and Gynecology, Bata Hospital, Zlin. METHODS: Endometrial samples (n=56) were obtained on days 5 and 7 after ovulation, progesterone addition, resp. Patients (n=14) included in the study had regular menstrual cycle, age under 40, BMI range 20-32 and basal FSH level range 5-9 IU/l. The collected samples were processed routinely and bcl-2 was estimated by indirect three level imunohistochemic method. RESULTS: We demonstrated the bcl-2 expression in all evaluated layers (epithelial surface, stroma, glands) of the mid-secretory endometrium in both cycles. No difference was found in bcl-2 expression between days 5 and 7 in the secretory phase of the natural cycle. In artificial cycles higher bcl-2 expression was found only in epithelial surface (p<0.05) between days 5 and 7. On day 7 higher bcl-2 expression was found in the artificial cycle in the endometrial epithelial surface (p<0.001) as well as in spongy layer (p<0.01) and compact layer (p<0.05) comparing to day 7 of the natural cycle. CONCLUSION: The bcl-2 expression in the mid-secretory endometrium is significantly higher in the cycle with estrogen-progesterone substitution comparing to the natural cycle. These changes were more significant in endometrial and glandular epithelium than in stromal cells. PMID- 15779299 TI - [Syndrome of pseudomyxoma peritonei--description of three cases and survey of the problem]. AB - OBJECTIVE: To describe personal experience with three heterogeneous cases of pseudomyxoma peritonei. Review of the literature concentrates on the pathogenesis and biologic spectrum of the rare syndrome. DESIGN: Three case reports. SETTING: Private Biopsy Lab s.r.o. and Sikl's Department of Pathology, Charles University and Faculty Hospital, Pilsen. METHODS AND RESULTS: Included are cases from Biopsy Lab s.r.o. and from personal consulting registry of Prof. Michal, Sikl's Department of Pathology, Pilsen. Clinical presentation of three cases of pseudomyxoma peritonei documented is in details including long follow up. Broad variability of histologic appearance associated with different Clinical outcome is demonstrated. Immunohistochemistry performed by streptavidin-biotin system (LSAB+, Dako) as detection system, and diaminobenzidine tetrahydrochloride as chromogen, employed three monoclonal antibodies--MUC2 (Novocastra, 1:400), cytokeratin 7 (Dako, 1:200) and cytokeratin 20 (Dako, 1:100). CONCLUSION: Pseudomyxoma peritonei is a clinical syndrome defined as presence of massive mucinous, viscous material in the peritoneal cavity, both floating and adhering to serosal surface (jelly-belly). The histologic examination should always follow with detailed and precise description of the epithelial component. The cytological and structural quality of the epithelium constitutes two basic forms with entirely different nature biology and prognosis. First and more frequent, so called disseminated peritoneal adenomucinosis, where primary low grade (benign) mucinous appendiceal tumor is almost constant finding, often recurs but displays favorable prognosis. Second, so-called peritoneal mucinous carcinomatosis is an extraordinary manifestation of peritoneal carcinosis following generalization of the gastrointestinal mucinous adenocarcinoma. Histopathogenesis in both types is thought to arise from MUC2 positive goblet cells in GI and ovarian involvement is secondary. A normal macroscopic finding on appendix or "uneventful" appendectomy in anamnesis is not unusual. PMID- 15779300 TI - [Cystic hypersecretory carcinoma: rare and potentially aggressive variant of intraductal carcinoma of the breast. Report of five cases]. AB - OBJECTIVE: To describe a poorly recognized histological and immunohistochemical features of cystic hypersecretory carcinoma of breast, including the overexpression of HER-2/neu protein and androgen receptors. SUBJECT: Review of clinical and pathological features of 5 cases of cystic hypersecretory carcinoma. SETTING: Private Bioptic Lab, Plzen and Department of Pathology, Charles University, Plzen. SUBJECT AND METHODS: Five cases of rare variant of intraductal carcinoma of the breast, so-called cystic hypersecretory carcinoma, are reported. The patients were females aged between 53 and 78 years (average 66.8 years). The size of the lesions ranged between 7 and 8 cm in largest dimension. RESULTS: In two cases, the development of high grade invasive ductal carcinoma was observed; in one additional case there was recurrence of high grade in situ carcinoma after three years. Strong overexpression of HER-2/neu protein was observed in three cases including those two with invasive component. Protein p53 was variably positive in all cases. Steroid receptor immunohistochemical study yielded variable results with only one case being positive for both estrogen and progesterone receptors. Interestingly, in most case (4/5) staining for androgen receptors was observed. CONCLUSION: The importance of correct diagnosis of this potentially aggressive lesion is emphasized. The differential diagnosis of breast lesions with predominantly cystic growth pattern is discussed in detail. PMID- 15779302 TI - [Problems with the ASCUS (atypical squamous cells of undetermined significance) in the Bethesda system]. PMID- 15779301 TI - [Differential diagnosis of inflammations of the vagina and external genitalia and possibilities and limits of microbiologic examination]. PMID- 15779303 TI - Prospects look so bright. AB - IHEEM is at a pivotal point in its development, says the institute's president Richard Nugent. He outlines to Health Estate Jouurnal's managing editor Nicholas Marshall how IHEEM is in a position to seize opportunities to become stronger through further widening its membership, and developing new roles for providing training and guidance. PMID- 15779304 TI - Green procurement meets energy targets. PMID- 15779305 TI - Managing change proactively. PMID- 15779306 TI - Pharmacies need fresh ideas dispensed. PMID- 15779307 TI - Infection control during construction. AB - "Construction" work requires special attention in healthcare facilities because infection caused by exposure to airborne dust contaminated with certain mould species can cause severe illness in patients with weakened immune systems. It is essential, therefore, that all construction work be assessed at the planning stage for potential impacts and, where required, be performed under carefully controlled conditions. Consideration must extend beyond dust generated by major projects to include dust that can become airborne during routine maintenance and minor renovation activ ties. A number of measures for controlling dust which are common practice in asbestos abatement and mould remediat on work can be applied to construction work carried out in healthcare facilities. PMID- 15779308 TI - Air flow disruption must be minimised. AB - Operating theatre lights are known to affect the air flow in ultra clean ventilation systems and it is important that specialist luminaires are designed to minimise the disruption of air flow. This paper explains the results of tests carried out during actual surgical interventions using reticulated operating theatre lights and examines the implications of new NHS recommendations on the future use of UCV ceilings. PMID- 15779309 TI - Dentin bonding--is there a future? PMID- 15779310 TI - Stress concentration in microtensile tests using uniform material. AB - PURPOSE: The objective of this study was to analyze the stress concentration factor (Kt) in specimens of uniform material with the most commonly used geometry (square hourglass) during microtensile tests using finite element analysis. Standardization is emphasized with the aim of obtaining the most representative nominal strength of the material. METHODS: Eighty cases were simulated using three-dimensional models, in which we varied the fixation of specimens in the jig (f = 1 or 2 sides), the height of this fixed region (h = 1 or 2.75 mm), the specimen width (D = 1.5, 2, 3, 4 or 5 mm), and the radius of curvature of the notch (r = 0.2, 0.5, 0.7 or 1 mm). The cross-sectional area (1 mm2) remained constant in all analyses. The stress concentration factor Kt (maximum tensile stress/nominal tensile stress) was calculated. RESULTS: A 150% difference was observed from the lowest Kt value (1.3) to the highest one (3.2). Results indicated that the radius of curvature is a very influential geometric parameter in microtensile strength tests (variation in Kt values up to 47.4%). For two-side fixed specimens, the Kt values varied from 3 to 4%, while the one-side fixed models resulted in variations from 11 to 15%. CONCLUSION: Variations in the specimen geometry and mode of load application can be responsible for part of the different strength values obtained in microtensile tests. The specimen fixation by two sides is a simple and easily performed method to reduce the stress concentration factor and its variations induced by specimen geometry and test assembly. PMID- 15779311 TI - Influence of crosshead speed on resin-dentin microtensile bond strength. AB - PURPOSE: The aim of this study was to investigate the influence of testing speed on the microtensile bond strength values of two adhesive systems to dentin. MATERIALS AND METHODS: A flat dentin surface was exposed after grinding the occlusal enamel of 10 human third molars. The adhesive systems (n = 5 each) Single Bond and Clearfil SE Bond were applied according to the manufacturers' instructions, and composite resin crowns (Z250) were constructed incrementally. After 24 h in distilled water at 37 degrees C, each tooth was longitudinally sectioned in both the x and y directions, under a cutting speed of 300 rpm, to obtain sticks with a cross-sectional area of 0.8 mm2. The sticks from each tooth were divided into 5 groups to be tested under crosshead speeds of 0.1, 0.5, 1.0, 2.0, and 4.0 mm/min. The fracture pattern was analyzed microscopically after debonding (400X). A two-way ANOVA having adhesive system (2 levels) and testing speed (5 levels) as main factors was used to compare the microtensile bond strengths. RESULTS: The two-way ANOVA did not detect statistically significant differences either for the interaction (p = 0.80) or for the crosshead speed (p = 0.69). Differences were only detected between adhesive systems (p = 0.001), with Single Bond showing higher bond strength values than Clearfil SE Bond. CONCLUSION: The crosshead speed used for microtensile bond strength tests does not influence the bond strength values, regardless of the adhesive system employed. PMID- 15779312 TI - Effect of surface preparation on microtensile bond strength of three adhesive systems to bovine enamel. AB - PURPOSE: The purpose of this study was to compare the microtensile bond strength (microTBS) of three adhesives to bovine enamel prepared with 600-grit silicon carbide paper, diamond rotary instrument, or carbide bur. MATERIALS AND METHODS: Bovine teeth (n = 36) were randomly divided into three treatment groups and bonded using a total-etch adhesive (Single Bond, 3M ESPE), a self-etching primer system (Clearfil SE Bond, Kuraray), or a self-etching adhesive (One-Up Bond F, Tokuyama). A 4-mm composite crown was built on the bonded surfaces and specimens were stored in water for one day at 37 degrees C. Specimens were sectioned into 0.7-mm-thick slabs, trimmed to a cross-sectional area of 1 mm2, and loaded to failure at a crosshead speed of 1 mm/min using a tabletop tester (EZ-Test, Shimadzu). Microtensile bond strength data were analyzed using ANOVA and Fisher's PLSD test (alpha = 0.05). RESULTS: The bond strength of each self-etching system was lower when the enamel was prepared using a diamond or carbide bur, rather than with 600-grit silicon carbide paper. Differences in microTBS between carbide and diamond-prepared surfaces were not significant. The surface preparation method did not affect the total-etch system. CONCLUSION: Different preparation instruments are unlikely to affect resin-enamel bond strengths. PMID- 15779313 TI - The role of cavity preparation and conditioning in the leakage of restorations. AB - PURPOSE: Restoration microleakage is thought to be determined by the method and location of cavity preparation, enamel etching, and dentin conditioning, as well as the restorative material. This study compared the microleakage of composite restorations placed in preparations cut with carbide and diamond burs and those treated with different bonding/conditioning agents. MATERIALS AND METHODS: Class V preparations (3 x 2 x 2 mm) were cut wholly in enamel or in enamel and cementum in 100 human premolars. Twenty teeth were prepared with carbide burs and the preparations etched and conditioned with Prime&Bond NT but not restored. A second set of 20 teeth had enamel-only preparations cut with carbides (n = 10) or diamonds (n = 10), and the preparations etched, conditioned (Prime&Bond NT) and restored with Prisma TPH. The other 60 teeth were divided into 3 groups of 20 teeth each with enamel-only (n = 10) or enamel/cementum preparations (n = 10). The 3 groups of teeth were conditioned with Optibond Solo, Clearfil SE Bond or Prompt-L-Pop prior to restoration with Prisma TPH. Two mm of root was resected from all teeth, pulpal tissue removed, and insulated copper wires inserted via the root canals to contact with the pulp chamber roof before the tooth-wire interfaces and root surfaces were sealed. The teeth were immersed in 0.9% NaCl and leakage assessed over 30 d by iR drop across a resistor in series with a DC source and stainless-steel counter electrode. RESULTS: Differences (p < 0.05) in leakage were found for enamel preparations cut with carbides and diamonds, and the relationship of leakage vs time was linear. Enamel/cementum preparations showed greater leakage, and the relationship of leakage vs time was sigmoidal. Conditioned-only preparations showed the same leakage as those conditioned and restored, while preparation leakage varied with the conditioning agent. CONCLUSION: Cavity preparation location, method of cutting, and the conditioning agent markedly affect leakage behavior. PMID- 15779314 TI - In vitro shear bond strength of self-etching adhesives in comparison to 4th and 5th generation adhesives. AB - PURPOSE: To determine the shear bond strength (SBS) of different established (Resulcin Aqua Prime & Monobond N: RA, Prompt L-Pop III: PLP) and experimental (AC-Bond: AC, AC-Bond + Desensitizer: ACD) self-etching adhesives in comparison to fourth (Optibond FL: FL) and fifth generation (Excite: EX, Gluma Comfort Bond: CB) adhesives. MATERIALS AND METHODS: All adhesives were applied on flat enamel and dentin surfaces and light cured following manufacturers' directions. Tetric Ceram A2 composite cylinders 3.5 mm in diameter and 2.0 mm in height were sheared off (1 mm/min) after thermocycling (5 to 55 degrees C, 5000x). The t-test (5% level, Bonferroni-correction) was used for statistical analysis. RESULTS: SBS in enamel: RA: 27.0+/-5.8 MPa, PLP: 15.9+/-3.4 MPa, AC: 28.1+/-4.4 MPa, ACD: 22.2+/ 4.1 MPa, FL: 33.2+/-3.2 MPa, EX: 30.5+/-5.1 MPa, CB: 30.1+/-3.7 MPa. SBS in dentin: RA: 25.8+/-5.7 MPa, PLP: 20.7+/-2.9 MPa, AC: 27.0+/-4.5 MPa, ACD: 20.7+/ 3.7 MPa, FL: 34.4+/-3.8 MPa, EX: 30.0+/-4.6 MPa, CB: 27.9+/-2.6 MPa. FL resulted in significantly (p < 0.002) higher SBS in enamel and dentin than RA, AC, ACD, and PLP, and in higher SBS to dentin than CB. In enamel and dentin, RA performed significantly superior to PLP, but was not different from AC and ACD. EX and CB were both on the same level of significance as AC and RA, but showed superior results to ACD and PLP (enamel and dentin). PLP resulted in significantly lower SBS values in enamel and dentin than all the other materials investigated, except ACD in dentin, to which it was equivalent. CONCLUSION: Resulcin Aqua Prime & Monobond N and AC-Bond were not significantly different than established 5th generation products. AC-Bond + Desensitizer and Prompt L-Pop have significantly different SBS from established 4th and 5th generation products. Future studies are required to investigate marginal integrity to determine if self-etching adhesives are an adequate alternative to one- and multi-bottle systems. PMID- 15779315 TI - Influence of activation mode of dual-cured resin composite cores and low viscosity composite liners on bond strength to dentin treated with self-etching adhesives. AB - PURPOSE: The objective of this study was to investigate the effect of light- or chemical initiation on the polymerization reaction of dual-cured composite core buildup materials (CM) with respect to their shear bond strength to dentin treated with self-etching adhesives (SE), in the presence or absence of a low viscosity composite liner (LI). MATERIALS AND METHODS: Two SEs (Clearfil SE Bond and Unifil Core Bond), their respective CMs (DC Core Paste and Unifil Core), and a LI (Protect Liner F) were tested. After removing the labial enamel surfaces of 8 bovine incisors, each flat dentin surface was sectioned into 4 pieces with similar surface areas (30 mm2). The dentin surfaces were wet abraded with 600 grit SiC paper and randomly divided into 8 groups. Experimental groups comprised the use of a SE and its respective CM with or without an intermediate application of the LI and with or without CM light activation. Three cylinders of CM (0.5 mm high and 0.75 mm in diameter) were applied to each bonded dentin surface (n = 12), using a tygon tubing mold. After water storage for 24 h, specimens were subjected to microshear testing. Data were statistically analyzed by two-way ANOVA and Tukey's test (5%). RESULTS: For both SEs, LI application and CM light activation showed significantly higher bond strengths than the groups in which an intermediate LI was not used or the CMs were only chemically cured. CONCLUSION: The bond strength of CM to dentin using SE is reduced if the LI and light activation are not employed. PMID- 15779316 TI - Effect of sodium hypochlorite on dentin bonding in primary teeth. AB - PURPOSE: The aim of this study was to evaluate the effect of sodium hypochlorite (NaOCl) on the shear bond strength (SBS) using three bonding systems in primary dentin. MATERIALS AND METHODS: Forty-five sound extracted primary molars were selected. The crowns were longitudinally sectioned, embedded in polystyrene resin, and flattened until a dentin surface was reached. The samples were assigned to 6 groups (n = 15): G1, Single Bond (SB); G2, NaOCl + SB; G3, Prime & Bond 2.1 (PB); G4, NaOCl + PB; G5, Clearfil SE Bond (CSE); G6, NaOCl + CSE. All the adhesive systems were applied according to the manufacturers' instructions, except for the application of 10% NaOCl solution for 60 s in groups 2, 4 (after acid etching), and 6 (before applying adhesive system). The composite resin was placed in increments in a mold and light cured for 20 s. The samples were stored in distilled water at 37 degrees C for 24 h and submitted to SBS testing with a crosshead speed of 0.5 mm/min. The failure sites were observed with SEM. The data were treated with ANOVA and Tukey's tests (p < 0.05). RESULTS: There was no statistically significant difference between the groups with or without treatment of the substrate with NaOCl, regardless the material used. The SBS averages in MPa (+/-SD) were: G1: 15.8(1.9)a; G2: 14.6(1.3)a; G3: 10.2(0.7)a; G4: 9.9(0.2)a; G5: 13.3(1.2)a; and G6: 10.7(1.0)a. There was a statistically significant difference between the materials (SB > or = CSE > or = PB). Mixed failure was the failure type most frequently observed for all groups. CONCLUSION: Dentin surface treatment with NaOCl did not affect the resin-dentin bonding strength in primary teeth. PMID- 15779317 TI - Microtensile bond strength of self-etching adhesives to enamel and dentin. AB - PURPOSE: To measure the microtensile bond strength to enamel and dentin of three self-etching adhesives in comparison with a total-etch two-step system as a control. MATERIALS AND METHODS: A total of 40 extracted human molars were stored in saline solution until use, then divided into 4 groups of 10 teeth (one group per adhesive system). Half of each of these groups underwent bond strength tests on enamel, and the other half was used for adhesion testing on dentin. The following experimental groups (n = 5) were then formed: E(1) Adper Prompt-L-Pop (AP, 3M ESPE) on enamel; E(2) Xeno CF II (X, Sanking Kogyo) on enamel; E(3) AdheSE (AS, Ivoclar-Vivadent) on enamel; E(4) Excite (EX, Ivoclar-Vivadent) on enamel; D(1) AP on dentin; D(2): X on dentin; D(3) AS on dentin; D(4) EX on dentin. Each tooth yielded 15 to 20 sticks about 0.9 x 0.9 mm in cross-sectional area for microtensile testing. Specimens were loaded in tension at a crosshead speed of 0.5 mm/minute, and bond strength at failure was calculated in MPa. A two way ANOVA was applied to test for significance of the differences among the groups. RESULTS: The bond strength values of Excite (the control) were significantly higher than those of the test products on enamel (42.92+/-4.8 MPa) and on dentin (45.80+/-5.79 MPa). The self-etching adhesives AdheSE (28.48+/-4.71 MPa) and Xeno CF II (27.22+/-2.74 MPa) revealed significantly stronger adhesion than Adper Prompt-L-Pop (20.16+/-2.07 MPa) on dentin. On enamel, all self-etching test materials performed similarly. The substrate did not appear to have a significant influence on adhesion, as each material reached comparable levels of bond strength on enamel and dentin. CONCLUSION: On both substrates the self etching adhesives tested performed significantly worse than did the total-etch system. PMID- 15779318 TI - Bonding of composite resin luting cement to fiber-reinforced composite root canal posts. AB - PURPOSE: The aim of this study was to compare bonding of composite resin luting cement to a fiber-reinforced composite (FRC) root canal post with either a cross linked or a semi-interpenetrating polymer network (IPN) polymer matrix. MATERIALS AND METHODS: Four different types of prefabricated FRC posts with a cross-linked polymer matrix and two types of FRC posts with a semi-IPN polymer matrix which were individually formed were tested. Serrated titanium posts served as the reference. An auto-polymerizing resin luting cement was used for cementing the posts into the holes of composite resin disks. The pull-out force was measured using a universal testing machine after the post/cement/disk system had been stored dry or thermocycled (5 degrees C/55 degrees C, 6000x) in water. The bonding sites of the posts were examined with SEM. RESULTS: The FRC posts with a semi-IPN polymer matrix gave significantly higher pull-out force values than the prefabricated FRC posts with a smooth surface and a cross-linked polymer matrix (p < 0.004). The highest pull-out force was obtained with serrated titanium posts. Both the type of FRC post and thermocycling had a significant effect on the pull-out force (n = 8, ANOVA p < 0.001 and p < 0.007, respectively). SEM photomicrographs confirmed the results. CONCLUSION: This study showed that FRC posts with a semi-IPN polymer matrix bonded better to composite resin luting cement than did prefabricated FRC posts with a cross-linked polymer matrix, although their pull-out force was not as high as that of the mechanically interlocked serrated titanium posts. PMID- 15779319 TI - Effect of ceramic surface treatment on the microtensile bond strength between a resin cement and an alumina-based ceramic. AB - PURPOSE: The objective of this study was to test the following hypothesis: the silica coating on ceramic surface increases the bond strength of resin cement to a ceramic. MATERIALS AND METHODS: In-Ceram Alumina blocks were made and the ceramic surface was treated: G1--sand-blasting with 110-microm aluminum oxide particles; G2--Rocatec System: tribochemical silica coating (Rocatec-Pre powder + Rocatec-Plus powder + Rocatec-Sil); G3--CoJet System: silica coating (CoJet-Sand) + ESPE-Sil. The ceramic blocks were cemented to composite blocks with Panavia F resin cement (under a load of 750 g/1 min). The cemented blocks were stored in distilled water at 37 degrees C for 7 days and sectioned along the x and y axes with a diamond disk. Samples with an adhesive area of ca 0.8 mm2 (n = 45) were obtained. The samples were attached to an adapted device for the microtensile test, which was performed in a universal testing machine (EMIC) at a crosshead speed of 1 mm/min. RESULTS: The obtained results were submitted to ANOVA and Tukey's test. Mean values of tensile strength (MPa) and standard deviation values were: (G1) 16.8+/-3.2; (G2) 30.6+/-4.5; (G3) 33.0+/-5.0. G2 and G3 presented greater tensile strength than G1. There was no significant difference between G2 and G3. All the failures took place at the ceramic/resin cement interface. CONCLUSION: The silica coating (Rocatec or CoJet systems) of the ceramic surface increased the bond strength between the Panavia F resin cement and alumina-based ceramic. PMID- 15779320 TI - Clinical evaluation of ceramic inlays and onlays made with two systems: a one year follow-up. AB - PURPOSE: The aim of this study was to evaluate the clinical performance of ceramic inlays and onlays made with two systems: sintered (Duceram [D], DeguDent) and pressable (IPS Empress [IPS], Ivoclar-Vivadent) after 1 year. MATERIALS AND METHODS: Seventy-four restorations - 37 IPS and 37 D - were cemented in 34 patients. Twenty-four premolars and 50 molars received Class II cavity preparations, totaling 28 onlays and 46 inlays. The restorations were evaluated by two independent investigators at baseline, 6 months, and 1 year, according to modified USPHS criteria. Fisher and McNemar statistical tests were employed to assess the survival rate. RESULTS: After one year, 100% of the restorations were assessed and all the restorations were considered clinically excellent or acceptable. Among the analyzed criteria, only the following received "Bravo" ratings: marginal discoloration: IPS (24.32%), D (13.51%); marginal integrity: IPS (10.81%), D (8.11%); color match: IPS (5.41%), D (5.41%); surface texture: IPS (2.70%), D (10.81%). No "Charlie" or "Delta" scores were given to the restorations. CONCLUSION: Among the analyzed criteria, only marginal discoloration differed statistically significantly from the results of the baseline examination for IPS Empress ceramic restorations (p = 0.008). No significant differences were found between the two ceramics. The two ceramic systems demonstrated excellent clinical performance after a period of 1 year. PMID- 15779322 TI - Neuronal organization of structures in the central part of the amygdaloid body of the brain. AB - The features of neurons in the central part of a amygdaloid body (AB) were studied and compared with published data on similar measures of cells in the anterior and posterior parts of the amygdaloid complex. Neurons were impregnated with silver nitrate using the Golgi method. Neurons were classified and measured as described by Leontovich (1978). Long-axon and short-axon neurons were identified and their quantitative characteristics were described. PMID- 15779321 TI - Distribution of calcium ions in the mixed synapses of Mauthner neurons in the goldfish in normal conditions, in exhaustion, and in conditions of adaptation to exhaustion. AB - The aim of this study was to investigate the structure of large myelinated club terminals of Mauthner neurons (MN) in the goldfish at different levels of functional activity and the distribution within these synapses of calcium ions as assessed using a modified pyroantimonate method. In intact preparations, calcium pyroantimonate precipitates were not seen in gap junctions (GJ) or desmosome-like contacts (DLC). Fibrillar bridges in DLC clefts were not contrasted. After natural stimulation, which induces long-term adaptation in MN, GJ showed electron dense precipitates lining the whole cleft. Granules and clumps of precipitate were also seen in DLC clefts, with intense deposition on bridges. Increases in calcium ion concentrations to and above the levels detectable by the pyroantimonate method are known to block electrotonic transmission; filamentous actin is known to conduct the electrotonic signal as a cation current. The staining of DLC bridges with calcium pyroantimonate is therefore evidence for an association between calcium ions and actin molecules, as DLC bridges consist of actin, i.e., we have obtained evidence for the functioning of bridges as electrotonic transsynaptic shunts at the moment of fixation. These data lead to the conclusion that DLC in mixed synapses, apart from the known adhesive functions, also have a communication function. This appears in extreme conditions, allowing the synapse to maintain or change its conductivity according to ongoing need. PMID- 15779323 TI - Selective involvement of opioids in the mechanisms of synapse-specific plasticity in the common snail during the acquisition of sensitization. AB - Studies on defensive behavior command neurons LP11 and RP11 in semi-intact common snail preparations were performed to investigate the effects of the opioid peptide met-enkephalin and the opioid antagonist naloxone on the effects of nociceptive sensitization. Application of nociceptive stimuli to the snail's head elicited marked reversible membrane depolarization along with depression of neuron responses to sensory stimulation during the short-term stage of sensitization and facilitation of responses in the long-term stage. Met enkephalin at a dose of 10 microM but not at a dose of 0.1 microM partially suppressed responses to nociceptive stimuli. Acquisition of sensitization during exposure to met-enkephalin at doses of 10 and 0.1 microM led to complete suppression of the facilitation of responses to tactile stimulation of the head. Facilitation of responses to chemical stimulation of the head and tactile stimulation of the foot in these conditions was similar to that of neurons in control sensitized animals. Acquisition of sensitization during exposure to met enkephalin and/or naloxone elicited selective suppression of facilitation of responses to chemical stimulation of the head but had no effect on facilitation of responses to tactile stimulation of the head and foot. Met-enkephalin and naloxone had no effect on the depression of neuron responses evoked by sensory stimulation in the short-term stage of sensitization. It is suggested that during the acquisition of sensitization in the common snail, opioids are involved in controlling the mechanism of nociception and in the mechanisms of selective induction of long-term plasticity of the synaptic inputs to command neurons activated by tactile and chemical stimulation of the animal's head. PMID- 15779324 TI - Effects of vibrissae removal on search accuracy in the water maze. AB - In two experiments, the vibrissae were clipped on either the left, the right, or both sides, and the rats were trained to find a submerged platform in the Morris water maze. In both experiments, animals without vibrissae on both sides or on the left consistently spent significantly more time in the "counter" area twice the platform diameter in size, surrounding the submerged platform, than intact controls. Counter preference was not as consistent across experiments in rats with right vibrissae removed. These results suggest that the vibrissae are required for proprioceptive location of the platform itself, but not for proximal search accuracy. Since ischemic damage to hippocampal CA1 pyramidal cells has also been reported to prolong counter search during training, the results support the suggestion that impaired hippocampal processing of proprioceptive information from the vibrissae may contribute to the increased latency to find the platform shown by ischemic rats. PMID- 15779325 TI - Age-related changes in the characteristics of evoked responses in field CA1 in rat hippocampus slices after deafferentation of the forelimb. AB - The effects of in situ transection of the median nerve in baby rats on the formation of the properties of synaptic transmission in the hippocampus were studied. Paired-pulse stimulation was used to identify the characteristics of short-term facilitation in living slices. Significant deviations in the time dynamics of facilitation from control values were seen in operated animals. Analysis showed that the decrease in the population spike amplitude, which we have described previously, may be due not only to functional causes, but also to morphological causes--elimination of neurons and their processes. It is suggested that a period of destructive changes is followed by the onset of a phase of intense formation of new synaptic contacts. The nature of the heterochrony of electrophysiological measures does not lead to any conclusion as to whether the maturation of intrahippocampal systems leads or lags in the experimental baby rats. It is suggested that hippocampal development along a new, abnormal pathway is induced. PMID- 15779326 TI - Aftereffects of microinjections of neurotensin into the substantia nigra of the brain on conditioned motor responses in rats with lesions to serotoninergic neurons. AB - The effects of neurotensin microinjections into the substantia nigra of the brain on the performance of motor reactions in response to positive (food reinforcement) and negative (unreinforced) conditioned signals were studied, along with the characteristic aftereffects of these microinjections in rats with lesions to serotoninergic neurons in the dorsal cervical nucleus. Lesions were produced by local administration of the selective neurotoxin 5,7 dihydroxytryptamine. Microinjections of neurotensin were found to weaken the disinhibitory effects of neurotoxin on extinction of intersignal responses and extinction of conditioned motor reactions in response to presentation of negative stimuli, but to have no marked effect on the performance of reactions evoked by positive conditioned signals. Changes persisted into subsequent experiments without peptide microinjections. Measurements of the level of preference of the animals for one quadrant of the experimental chamber showed that operated rats avoided occupying the test quadrant and preferred to locate themselves in the opposite quadrant. Neurotensin released this effect of the neurotoxin. These results lead to the conclusion that these behavioral effects of neurotensin are associated with its normalizing influences on the motivational-emotional state of the animals with lesions to serotoninergic neurons and may also be due to the formation in the animals of a contextual conditioned emotional state, which aids optimization of the adaptive functions of the brain. PMID- 15779327 TI - Evoked brain potentials in adolescents in normal conditions and in attention deficit during solution of tasks requiring recognition of short-duration acoustic stimuli. AB - This study compares the effectiveness of the frequency-based recognition of short acoustic stimuli in groups of adolescents with attention deficit and normal measures of attention in conditions of the standard "oddball" paradigm. Stimuli of duration 50 msec yielded insignificant intergroup differences, though adolescents with attention deficit discriminated signal of duration 11 msec significantly worse. These showed significant differences in evoked brain potentials even with standard stimuli, with a significantly greater amplitude for N2b waves and a decreased P3b component. Evoked potentials obtained in response to the deviant stimulus were characterized by a P3b wave of reduced amplitude in the group with attention deficit. These data provide evidence that adolescents with attention deficit show defined abnormalities in the processing of acoustic sensory information at the cortical level. PMID- 15779328 TI - Effects of selective visual attention in the parietal and temporal areas of the human cortex using evoked potential data. AB - Studies of 11 young subjects addressed the analysis of evoked potentials in the parietal and temporal areas to sequential presentation of visual symbols on the left and right sides; symbols were squares and circles and were observed passively and in conditions of selective attention to target stimuli presented to the subjects in random order and requiring rapid and precise motor responses. Comparison of monopolar evoked potentials in leads P3, P4, T3, T4, T5, and T6 with bipolar potentials in leads P3-T3, P3-T5, P4-T4, and P4-T6 in conditions of passive and selective attention showed that voluntary attention was accompanied by significant rearrangements in evoked activity in the parietal and temporal areas. This was evident as: 1) an increase in correlations between evoked potentials in the parietal and temporal areas; 2) stabilization of monopolar evoked potentials over time, i.e., increases in the correlations of sequential evoked potentials, in both associative visual areas; 3) stabilization of bipolar parietal-temporal evoked potentials in terms of increases in their sequential correlations. It is suggested that selective attention facilitates linked activity of the two associative areas, which is needed for performance of visual selection. PMID- 15779329 TI - Actions of pulsed ultra-broadband electromagnetic irradiation on the EEG and sleep in laboratory animals. AB - Irradiation of animals with ultrashort impulses of ultra-broadband magnetic irradiation with an impulse repetition frequency of 6 Hz for 1 h induced changes in the spectral composition of cerebral cortex electrical activity in rats, measured over the 5 min immediately after irradiation, as compared with controls. In particular, there was suppression of frequencies close to the impulse sequence frequency, along with a decrease in interhemisphere coherence. Continuous recording of polygrams for 22 h from rabbits after irradiation revealed a "delayed" effect--a significant increase in paradoxical sleep, starting 16 h after the end of irradiation and persisting to the end of the recording period. It is suggested that irradiation has a direct action both on the mechanisms of generation of the theta rhythm (septohippocampal) and on the system controlling circadian rhythms (the suprachiasmatic nucleus-epiphysis system). PMID- 15779330 TI - Hypothalamic monoamines in cold stress on the background of changes in the activity of the nitric oxide system. AB - The effects of the NO donor sodium nitroprusside and the NO synthase blocker L omega-N-nitroarginine (LNA) on body temperature, hypothalamic monoamines, and plasma corticosterone in conditions of cooling were studied in Male Wistar rats. Reductions in body temperature on cooling, both after administration of sodium nitroprusside and LNA, were no different from those seen without treatment. The basal corticosterone level after treatment with sodium nitroprusside increased from 5.3 +/- 2.2 to 29.1 +/- 1.8 microg%. Cooling led to a multiple increase in corticosterone levels in all animals, both in control conditions and after treatment with sodium nitroprusside and LNA. Sodium nitroprusside significantly decreased the basal hypothalamic noradrenaline level, by 37%. Cooling of the animals in these conditions led to an additional drop in the noradrenaline level. Noradrenaline levels 48 h after cold stress applied to animals cooled after treatment with LNA or sodium nitroprusside were significantly higher than in those cooled without treatment. No changes in serotonin and 5-hydroxyindoleacetic acid levels were seen in these experiments. The basal dihydroxyphenylacetic acid and dopamine levels increased after treatment with sodium nitroprusside, by 379% and 239% respectively. No dopamine response to cold was observed, though the dihydroxyphenylacetic acid level in the control group and animals treated with LNA increased. Thus, cold stress did not reveal differently directed directions for the actions of the NO donor and the NO synthase blocker, as seen with other types of stress. PMID- 15779331 TI - Effects of the C-terminal peptide of the alphaS subunit of the G protein on the regulation of adenylyl cyclase and protein kinase A activities by biogenic amines and glucagon in mollusk and rat muscles. AB - The C-terminal parts of the a subunits of heteromeric G proteins play an important role in the functional linkage of G proteins with receptors of the serpentine type. The present report describes studies of the effects of the C terminal octapeptide 387-394 of the alphaS subunit of the mammalian G protein on the transmission of the hormonal signal via the hormone-sensitive adenylyl cyclase signal system, whose major components are receptors of the serpentine type, G proteins, and the enzymes adenylyl cyclase and protein kinase A. The peptide synthesized here, 387-394 amide (10(-7) - 10(-4) M), dose-dependently decreased adenylyl cyclase and protein kinase A activities stimulated by serotonin and glucagon in smooth muscle from the freshwater bivalve mollusk Anodonta cygnea and by the beta agonist isoproterenol in rat skeletal muscle. At a concentration as low as 10(-7) M, the peptide released potentiation of the stimulatory effects of hormones on adenylyl cyclase activity due to the non hydrolyzable guanine nucleotide analog Gpp[NH]p. At the same time, it had almost no effect on the stimulation of adenylyl cyclase activity by non-hormonal agents (NaF, Gpp[NH]p, and forskolin). The inhibitory effects of hormones on adenylyl cyclase and protein kinase A activities persisted in the presence of the peptide. Our data demonstrate the importance of the C-terminal part of the alphaS subunit of the stimulatory G protein for its functional linkage with receptors of the serpentine type and throw light on the molecular mechanisms of the interactions between G proteins and receptors. PMID- 15779332 TI - Function of the hypothalamo-hypophyseal-adrenal system in mice with ectopic hyperproduction of the agouti protein. AB - The agouti protein is known to compete with the melanocortin hormones (ACTH, melanocyte-stimulating hormone) at melanocortin receptors, which in turn are involved in controlling the central and peripheral components of the hypothalamo hypophyseal-adrenal system. The aim of the present work was therefore to assess the effects of the dominant mutation Agouti yellow (A(y)/a), which induces ectopic hyperproduction of the agouti protein and yellow coat color, on the function of the hypothalamo-hypophyseal-adrenal system. Experiments were performed on male A(y)/a mice of the C57BI/6J line. Controls consisted of mice of the same line bearing the recessive mutation nonagouti (a/a), which leads to the absence of agouti protein and black coat color. The experimental results showed that mice with different agouti genotypes had identical basal corticosterone levels, though yellow mice, as compared with black mice, had increased corticosterone levels after restriction stress (p < 0.02), along with decreased stress reactivity after treatment with dexamethasone (p < 0.0007), and increased adrenal sensitivity to small doses of activity, both in vitro and in vivo. PMID- 15779333 TI - Anxiety in females induced by long-lasting psychoemotional influences. AB - The experiments reported here addressed the behavior of female mice after long lasting psychoemotional influences consisting of daily placing of females in the presence of intermale confrontations, the females being on the other side of a transparent perforated partition; females were subsequently placed in the territory (litter) in which the conflict had taken place. These conditions induced anxiety in the females, detected in the elevated cross maze test. In the absence of any marked prodepressant effect, females were more passive in the Porsolt test. There was also an increase in the time spent by females close to the partition both 5 min before the start of the conflict and during the intermale conflict. PMID- 15779334 TI - Psychoemotional manifestations in hippocampectomized rats. AB - The effects of lesioning of the dorsal hippocampus on the psychoemotional state of Wistar rats were studied. Hippocampal lesions did not affect the learning process, but affected mainly the pattern of psychoemotional manifestations in all animals regardless of the typology of higher nervous activity. All animals showed reductions in the extremes of types of passive and active stress reactions. Individual effects depended on the behavioral phenotypes of the animals. Hippocampal lesions in rapidly learning rats (10%) led to sharp decreases in irritability (active/passive = 0.35/0.5 instead of 1.0/0.4), while passive defensive manifestations were not altered. In slowly learning animals (30%), hippocampal lesions were not accompanied by qualitative changes in the patterns of psychoemotional manifestations, while non-learning rats (60%) showed decreases in passive and increases in active manifestations (active/passive = 0.45/1.4 instead of 0.3/1.9). It is suggested that the hippocampus is involved in producing psychoemotional reactions, individual patterns of which depend on the morphofunctional characteristics of the system responsible for organizing the psychoemotional state. PMID- 15779335 TI - Characteristics of evoked potentials of the human brain during recognition of short acoustic stimuli of different frequencies. AB - We report here studies of the efficiency of recognition of short acoustic signals (11-30 msec) whose presentation did not produce mismatch negativity in evoked potentials in the human brain. Nonetheless, stimuli could still be recognized efficiently in these conditions; processing negativity disappeared from evoked potentials and the efficiency of recognition was significantly worsened on presentation of the shortest stimuli (11 msec). It is suggested that processing negativity also has a critical duration for acoustic signals, though this is significantly shorter than the minimum duration of stimuli needed for generating mismatch negativity. PMID- 15779338 TI - [Future increase in number of Japanese diabetics]. PMID- 15779336 TI - The significance of individual resistance to hypoxia for correction of the consequences of craniocerebral trauma. AB - Experiments on rats showed that the individual resistance of the body to acute hypoxia is of decisive importance in the early recovery period after mechanical craniocerebral trauma. I.p. administration of ethomersol (25 mg/kg) during the three days following trauma decreased behavioral impairments in rats with different levels of resistance to acute hypoxia, restored the structure of individual behavior, and prevented metabolic disturbances in the brain. The results led to the conclusion that the antioxidant ethomersol is effective in hypoxia due to craniocerebral trauma. PMID- 15779337 TI - The effects of the D1 receptor antagonist SCH-23390 on individual and aggressive behavior in male mice with different experience of aggression. AB - The dopaminergic systems of the brain are known to be involved in the mechanisms of aggression. The present report describes studies of the effects of acute administration of the dopamine D1 receptor antagonist SCH-23390 (0.1 mg/kg, i.p., over 30 min) on the individual and aggressive behavior of male mice of the line C57BL/6J with different experience of aggression. A group of animals with no previous experience of aggression responded to administration of this agent with decreases in direct aggression (attacks), though the total time of hostile behavior, i.e., the sum of the durations of attacks, aggressive grooming, and scattering of foreign litter, showed no change. The agent had no effect on the aggressivity of animals with 20 days of experience of agonistic confrontations. The discussion addresses the possible development of pharmacological densensitization of dopamine D1 receptors in aggressive males in response to prolonged activation of the dopaminergic systems in conditions of repeated experience of aggression, as demonstrated previously. PMID- 15779339 TI - [Borderline type, impaired glucose tolerance (IGT), and impaired fasting glucose (IFG): their concepts, classification and significance]. PMID- 15779340 TI - [Impaired glucose tolerance (IGT): current understanding and future research directions]. PMID- 15779341 TI - [IFG and IGT in Europe and America: the present and prospective]. PMID- 15779342 TI - [Clinical relevance of IGT in the DECODA Study]. PMID- 15779343 TI - [Definition and diagnosis of IFG, IGT and borderline type]. PMID- 15779344 TI - [Primary prevention of diabetes--importance of early diagnosis and interventions in IGT & IFG]. PMID- 15779346 TI - [Natural history of glucose intolerance groups]. PMID- 15779345 TI - [Effect of impaired fasting glucose and impaired glucose tolerance on the progression of diabetic macroangiopathy]. PMID- 15779347 TI - [International comparison of morbidity in diabetes and IGT]. PMID- 15779348 TI - [Epidemiologic investigation on the incidence of diabetes mellitus from impaired glucose tolerance in population based study]. PMID- 15779349 TI - [The prevalence of impaired glucose tolerance in life-style related disease]. PMID- 15779350 TI - [The internal and external cohort studies about impaired glucose tolerance]. PMID- 15779352 TI - [Pathogenic mechanism of type 2 diabetes mellitus from impaired glucose tolerance/borderline type and its reversibility]. PMID- 15779351 TI - [Etiologies of impaired glucose tolerance and impaired fasting glycemia]. PMID- 15779353 TI - [Mechanism of progression of atherosclerosis in patients with impaired glucose tolerance (IGT)]. PMID- 15779354 TI - [Insulin release impairment in IGT]. PMID- 15779355 TI - [Molecular mechanism of insulin resistance]. PMID- 15779356 TI - [Impaired hepatic glucose metabolism]. PMID- 15779357 TI - [Disorder of glucose metabolism in skeletal muscle]. PMID- 15779358 TI - [Impaired glucose-uptake in adipocytes]. PMID- 15779359 TI - [Postprandial hyperglycemia]. PMID- 15779360 TI - [Insulin secretion and insulin sensitivity in subjects with impaired fasting glucose]. PMID- 15779361 TI - [Type 2 diabetes and genetic variations of calpain 10 gene]. PMID- 15779362 TI - [S20G mutation of amylin gene--amyloid diabetes due to S20G amylin gene mutation]. PMID- 15779363 TI - [PPARgamma gene Pro12Ala polymorphism in diabetes mellitus]. PMID- 15779364 TI - [HNF-1alpha G319S mutation in Oji-Cree type 2 diabetes]. PMID- 15779365 TI - [I164T polymorphism of adiponectin gene in diabetes]. PMID- 15779366 TI - [Beta3-adrenergic receptor gene polymorphism in diabetes]. PMID- 15779367 TI - [Fatty acid binding protein-2 (FABP2) gene polymorphism in diabetes]. PMID- 15779368 TI - [TNF-alpha gene polymorphism, insulin resistance and type 2 diabetes in humans]. PMID- 15779369 TI - [Genetic variations of urotensin II gene in type 2 diabetes]. PMID- 15779370 TI - [Polymorphisms in ACE gene deletion/insertion in type 2 diabetes]. PMID- 15779371 TI - [Polymorphisms of uncoupling protein-1 gene in type 2 diabetes]. PMID- 15779372 TI - [Risk factors in pathogenesis of diabetes: trends in food intake]. PMID- 15779373 TI - [Factors influencing occurrence of diabetes: nutrient intakes (fat, animal protein and others)]. PMID- 15779374 TI - [Factors influencing occurrence of diabetes: eating behavior]. PMID- 15779375 TI - [Amount of exercise necessary for prevention of diabetes (amount of leisure-time exercise)]. PMID- 15779376 TI - [Effect of daily exercise in prevention of impaired glucose tolerance]. PMID- 15779377 TI - [Effect of alcohol consumption on glucose metabolism]. PMID- 15779378 TI - [Pathophysiological conditions progressing from impaired glucose tolerance: type 2 diabetes mellitus]. PMID- 15779379 TI - [Pathophysiological conditions progressing from impaired glucose tolerance: high blood pressure]. PMID- 15779380 TI - [Significance of visceral fat syndrome in pathogenesis of impaired glucose tolerance]. PMID- 15779381 TI - [Pathophysiological conditions progressing from impaired glucose tolerance: atherosclerosis]. PMID- 15779382 TI - [Pathophysiological conditions progressing from impaired glucose tolerance: coronary artery disease (ischemic heart disease)]. PMID- 15779383 TI - [Pathophysiological conditions progressing from impaired glucose tolerance: stroke (cerebral infarction)]. PMID- 15779384 TI - [Obesity and the metabolic syndrome]. PMID- 15779385 TI - [Hyperuricemia as a unique marker for metabolic syndrome]. PMID- 15779386 TI - [Glucose intolerance in endocrinopathies]. PMID- 15779387 TI - [Glucose tolerance in renal failure]. PMID- 15779388 TI - [Glucose intolerance after renal transplantation]. PMID- 15779389 TI - [Impaired glucose tolerance in chronic liver disease]. PMID- 15779390 TI - [Impaired glucose tolerance in nonalcoholic fatty liver disease]. PMID- 15779392 TI - [Impaired glucose tolerance in patients with ischemic heart diseases]. PMID- 15779391 TI - [Impaired glucose tolerance in dumping syndrome]. PMID- 15779393 TI - [Genetic syndromes associated with glucose intolerance]. PMID- 15779394 TI - [Glucose intolerance caused by interferon therapy]. PMID- 15779395 TI - [Effects of Ca channel blockers on glucose metabolism and insulin sensitivity]. PMID- 15779396 TI - [Glucocorticoid-induced diabetes]. PMID- 15779397 TI - [Glucose intolerance with antipsychotic drugs]. PMID- 15779398 TI - [Glucose intolerance induced by tacrolimus (FK506), cyclosporin A]. PMID- 15779400 TI - [Impaired glucose tolerance in the elderly--prevalence, pathogenesis and management]. PMID- 15779399 TI - [Glucose intolerance due to percutaneous ethanol injection therapy]. PMID- 15779401 TI - [Impaired glucose tolerance in children and adolescents]. PMID- 15779402 TI - [Glucose intolerance during pregnancy]. PMID- 15779403 TI - [Diagnosis and tests for impaired glucose tolerance and borderline type]. PMID- 15779404 TI - [Significance of insulin measurement in subjects with impaired glucose tolerance or type 2 diabetes]. PMID- 15779405 TI - [Screening for type 2 diabetes and impaired glucose tolerance]. PMID- 15779406 TI - [Oral glucose tolerance test (OGTT) for diagnosis of diabetes mellitus]. PMID- 15779407 TI - [Fasting plasma glucose for diagnosis of diabetes mellitus]. PMID- 15779408 TI - [Analysis of HbA1c and glycoalbumin for diagnosis of impaired glucose tolerance]. PMID- 15779409 TI - [Plasma anhydro-glucitol as a blood sugar control marker for impared glucose tolerance]. PMID- 15779410 TI - [Combined use of fasting plasma glucose and HbA1c (as a screening test)]. PMID- 15779411 TI - [2h-CPR/2h-PG in 75g-OGTT]. PMID- 15779412 TI - [Detection of glucose intolerance by urinary myo-inositol]. PMID- 15779414 TI - [Efficacy of IMT as the quantitative measure of carotid artery atherosclerosis]. PMID- 15779413 TI - [Microalbuminuria in patients with impaired glucose tolerance]. PMID- 15779415 TI - [The cutoff point of visceral fat area as a goal for improving risk factors of arteriosclerosis in patients with glucose intolerance]. PMID- 15779416 TI - [Treatment for patients with impaired glucose tolerance]. PMID- 15779417 TI - [Selection of oral antidiabetic drugs]. PMID- 15779418 TI - [Characteristic of metformin for treatment of impaired glucose tolerance]. PMID- 15779419 TI - [Effects of nateglinide in impaired glucose tolerance subjects]. PMID- 15779420 TI - [Effects of mitiglinide in treatment of impaired glucose tolerance]. PMID- 15779421 TI - [Effects of acarbose, alpha-glucosidase inhibitor in treatment of impaired glucose tolerance]. PMID- 15779422 TI - [Effects of voglibose, alpha-glucosidase inhibitor in treatment of impaired glucose tolerance]. PMID- 15779423 TI - [Effect of pioglitazone, one of TZDs, on IGT-patients]. PMID- 15779424 TI - [Diabetes prevention program (metformin)]. PMID- 15779425 TI - [The Study TO Prevent Non-Insulin Dependent Diabetes Mellitus trial (acarbose)]. PMID- 15779426 TI - [NAVIGATOR trial (nateglinide)]. PMID- 15779427 TI - [Japan Diabetes Prevention Program]. PMID- 15779428 TI - [Malmo Feasibility Study (Sweden)]. PMID- 15779429 TI - [The Da Qing IGT and Diabetes study]. PMID- 15779430 TI - [Diet therapy for impaired glucose tolerance]. PMID- 15779431 TI - [Nutrition guidance for elderly glucose intolerance]. PMID- 15779432 TI - [Review of exercise in prevention of diabetes]. PMID- 15779433 TI - [Exercise therapy for obese impaired glucose tolerance patients]. PMID- 15779434 TI - [Cognitive behavior therapy for impaired glucose tolerance]. PMID- 15779436 TI - [Treatment of hypertensive patients with impaired glucose tolerance (IGT)]. PMID- 15779435 TI - [Therapies for newly-onset diabetic patients]. PMID- 15779437 TI - [Dyslipidemia management in patients with impaired glucose tolerance]. PMID- 15779438 TI - [Treatment for coronary artery disease patients with impaired glucose tolerance]. PMID- 15779439 TI - [Treatment of cerebral infarction patients with IGT]. PMID- 15779440 TI - [Treatment of obese patients with IGT]. PMID- 15779441 TI - [Perioperative management for patients with impaired glucose tolerance (IGT)]. PMID- 15779443 TI - [Prevention of type 2 diabetes mellitus by changing life-styles among high risk persons: the Diabetes Prevention Program in Hiroshima (DPPH)]. PMID- 15779442 TI - [Diabetic campaign in Toyama: framework and program for organized cooperation in regional diabetes care and prevention]. PMID- 15779444 TI - [Management of IGT education program at a hospital and its primary prevention effect on diabetes mellitus]. PMID- 15779445 TI - [An approach to the prevention of type 2 diabetes in industrial healthcare]. PMID- 15779446 TI - [The global burden of impaired glucose tolerance: present and future strategy]. PMID- 15779447 TI - [The 2002 National Nutrition Survey and current status of obesity, dietary intake and habitual exercise among Japanese]. PMID- 15779448 TI - [World wide pandemic of childhood and adolescence type 2 diabetes mellitus and in Japan]. PMID- 15779449 TI - [Development of health-risk appraisal in impaired glucose tolerance]. PMID- 15779450 TI - [Diabetic retinopathy]. PMID- 15779451 TI - [Diabetic nephropathy]. PMID- 15779452 TI - [Diabetic neuropathy]. PMID- 15779453 TI - [Arteriosclerosis in diabetes]. PMID- 15779454 TI - [Decreased serum levels of adiponectin as a risk factor for development of type 2 diabetes]. PMID- 15779456 TI - [Evaluation of health promotion program for patients with glucose intolerance]. PMID- 15779455 TI - [Is hyperfattyacidemia related to the occurrence of diabetes?]. PMID- 15779457 TI - [Mechanisms of physical exercise for the treatment of mild diabetic patients]. PMID- 15779458 TI - [Cost-effectiveness of the diabetes prevention program for patients with IGT]. PMID- 15779459 TI - Evaluation of the implantation position of mini-screws for orthodontic treatment in the maxillary molar area by a micro CT. AB - The interalveolar septum between the upper first molar and the second premolar of the separated human maxillary bone was three-dimensionally observed by micro CT to evaluate the appropriate mini-screw type implant placement position by considering the relationship between the tooth roots and the maxillary sinus. After taking micro CTs of 5 human maxillary bones, horizontally sectioned images of the interalveolar septum area 2, 4, 6, 8, 10, and 12 mm deep from the crest of the alveolar ridge were reconstructed by three-dimensional reconstruction software. The bucco-lingual and mesio-distal lengths and area in each sectioned interalveolar septum were measured using digital image measurement software. Using the results, the interalveolar septum area between the upper first molar and the second premolar approximately 6-8 mm deep from the alveolar crest in the tooth root apical direction was determined to be the safest position for mini screw implantation. Furthermore, lateral implantation from the palatal side was deduced to be the safest approach. PMID- 15779460 TI - Comparison of short-term in vitro fluoride release and recharge from four different types of pit-and-fissure sealants. AB - OBJECTIVE: The purpose of this in vitro study was to assess the effects of four commercial fluoride-containing pit-and-fissure sealants on caries prevention. MATERIALS AND METHODS: Four sealants containing fluoride, Fuji III, Fuji III LC (GC Co., Tokyo), Teethmate F-1 (Kuraray Medical Co., Osaka) and Helioseal F (Vivadent Co., Liechtenstein) were used to investigate fluoride release and recharge. Disk-shaped specimens prepared from each material were immersed in distilled water at a temperature of 37 degrees C. After seven days, acidulated phosphate fluoride solution (APF) was applied to each specimen, and it was then again immersed in distilled water for 14 days. We then determined how much fluoride had been released into the immersing water. Fuji III LC was used with APF solution to investigate the fluoride uptake. RESULTS: Fuji III had the highest fluoride release, and Fuji III LC had the highest fluoride recharge. Helioseal F and Teethmate F-1 had almost no fluoride recharge. Fuji III LC/APF had a higher fluoride uptake to enamel than Fuji III LC. CONCLUSIONS: These results suggest that GIC-sealants in the oral cavity can serve as a fluoride reservoir and contribute to retaining a low fluoride level in oral fluids, thereby preventing caries. PMID- 15779461 TI - Approach to obstructive sleep apnea syndrome at Tokyo Dental College, Ichikawa General Hospital. AB - In this communication, we report the current status of OSAS (Obstructive Sleep Apnea Syndrome) in the southern region of Higashikatsushika around Ichikawa City, our effort to improve patient QOL as well as to establish diagnostic and therapeutic methods, and the results of a comparison of therapeutic options with the focus on improvement of compliance by using nCPAP (nasal continuous positive airway pressure). We examined 112 patients who visited the Otolaryngology Department at Tokyo Dental College, Ichikawa General Hospital, with the chief complaint of nocturnal snoring or sleep apnea from January 2001 to April 2003 and underwent all-night PSG (polysomnography). Based upon the results of these all night PSGs, 89 and 23 patients were diagnosed as having OSAS and simple snoring, respectively. Using the AHI classification of severity, 58 and 31 patients were assessed as having severe OSAS and mild OSAS, respectively. (1) nCPAP was tried in 61 patients, and 39 patients (63%) were able to continue it. After the introduction of nCPAP, surgery was performed in 18 patients (30%). As a result, weaning from nCPAP was successfully achieved in 10 cases, compliance with nCPAP was improved in six cases, alleviation of symptoms (decreased pressure) was seen in one case, and aggravation was noted in one case. In addition, four patients (7%) unilaterally discontinued nCPAP. (2) Surgery was performed in 34 patients, and 18 of them had surgery after nCPAP was tried. (3) We asked the dental department to make OAs (oral appliances) for 31 patients but seven of them did not attend the department, so a total of 24 patients used OAs. Fourteen patients (58%) were able to tolerate an OA for 3 months or more. Based on these results, we are hoping to achieve a better control of OSAS by combining nCPAP and other modalities. PMID- 15779462 TI - Endothelin-1-induced depolarization and hyperpolarization in submandibular ganglion neurons. AB - The effects of endothelin-1 were studied in vitro on neurons in the hamster submandibular ganglion, using the intracellular microelectrode technique. Endothelin (1 microM) caused a depolarization (5.5 +/- 1.2 mV) followed by a hyperpolarization (8.5 +/- 2.8 mV) of the membrane potential. Membrane conductance was increased during the endothelin-induced depolarization and was decreased during the endothelin-induced hyperpolarization. The endothelin-induced depolarization was depressed (mean 43.6%) in a Krebs solution containing zero calcium and high magnesium. The results suggested that the predominant component of the depolarization was mediated by calcium ions. The calcium-insensitive component of depolarization was carried by chloride ions. Endothelin-induced slow rhythmic hyperpolarizations were probably induced by a decrease in chloride ion conductance. PMID- 15779463 TI - Ironies of success: a new history of the American health care "system". AB - The context for contemporary research and policy is set through a theoretically informed history of the modern American health care system that draws on the concept of countervailing powers and Fligstein's theory of control. In this context, the papers of this special issue are then introduced. PMID- 15779464 TI - Why the United States has no national health insurance: stakeholder mobilization against the welfare state, 1945--1996. AB - The United States is the only western industrialized nation that fails to provide universal coverage and the only nation where health care for the majority of the population is financed by for-profit, minimally regulated private insurance companies. These arrangements leave one-sixth of the population uninsured at any given time, and they leave others at risk of losing insurance as a result of normal life course events. Political theorists of the welfare state usually attribute the failure of national health insurance in the United States to broader forces of American political development, but they ignore the distinctive character of the health care financing arrangements that do exist. Medical sociologists emphasize the way that physicians parlayed their professional expertise into legal, institutional, and economic power but not the way this power was asserted in the political arena. This paper proposes a theory of stakeholder mobilization as the primary obstacle to national health insurance. The evidence supports the argument that powerful stakeholder groups, first the American Medical Association, then organizations of insurance companies and employer groups, have been able to defeat every effort to enact national health insurance across an entire century because they had superior resources and an organizational structure that closely mirrored the federated arrangements of the American state. The exception occurred when the AFL-CIO, with its national leadership, state federations and union locals, mobilized on behalf of Medicare. PMID- 15779465 TI - The misalignment of institutional "pillars": consequences for the U.S. health care field. AB - This paper uses an institutional perspective (Scott, 1994; 2001; DiMaggio and Powell, 1991) to analyze the history and current state of the American health care field in terms of the alignment of its normative, cognitive, and regulatory elements. I depict the relation between institutional elements in each of three historical eras of the health care field (Scott et al. 2000): the era of professional dominance (1945--1965), the era of federal involvement (1966--1982), and the era of managerial control and market mechanisms (after 1983 to the present). I argue that a weakening of alignment between these elements, beginning in the 1970s and increasing in the 1980s, led to consumer and provider dissatisfaction with managed care, and that the state of the field in the beginning of the 21st century suggests that a new era is emerging with renewed alignment between normative beliefs and values, cognitive models, and regulation. Implications for the future of health care and institutional theory are discussed. PMID- 15779466 TI - Unfamiliar tasks, contested jurisdictions: the changing organization field of medical practice in the United States. AB - This article proposes that Andrew Abbott's approach to the professions in his System of Professions can usefully be adapted to the study of organizational populations in an organization field. The key phenomena to study using this approach are conflicts over the definition of tasks and over the establishment of jurisdiction over these tasks by particular organizational populations. The following article uses this "tasks and jurisdictions " approach to describe the evolution of the field of medical practice in the United States during the era of managed care. Large employers and the federal and state governments have created two new and unfamiliar tasks--controlling costs and improving quality for defined populations of patients--and assigned jurisdiction over them to health plans. However, these tasks are not well understood, and health plan jurisdiction over them is not accepted by the public or by physicians. PMID- 15779467 TI - The rise and fall of managed care. AB - The managed care backlash is analyzed as a collective behavioral response led by attacks from threatened professional, provider, and special interest communities. Central to the backlash was the middle class's repudiation of explicit rationing at the point of service adopted by Health Maintenance Organizations (HMOs) and other managed care plans. Americans are accustomed to choice and autonomy in their health care utilization and reacted negatively to restrictions. Assisted by negative anecdotes in the media and allegations of the denial of needed services, opponents built a negative picture of managed care often inconsistent with the empirical evidence. They succeeded in arousing public anxiety that appropriate health care might not be available when people most needed it. The private centralization of large health plans made them an easy target for the media, politicians, and provider groups in opposition. These attacks and regulatory initiatives succeeded in diluting strong utilization management controls contributing to current difficulties of containing large health cost increases. PMID- 15779468 TI - The U.S. long-term care field: a dialectic analysis of institution dynamics. AB - This paper presents an institutional analysis of organizational change and inertia in the US. field of long-term care. We employ a dialectic approach to concentrate on the contest between interests aligned with two templates of organization (archetypes) that draw from distinctive sets of beliefs and values (interpretive schemes) to specify appropriate structural forms, roles, and resource distributions. It is shown that the long-term care field was historically characterized by a nursing home archetype which legitimates the provision of care in residential facilities under the control of medical professionals. After a century of reformers' efforts to build legitimacy and resource support for alternative home and community-based services that maintain consumers' independence, the field now accommodates both the nursing home archetype and a home and community-based services archetype. While this new institutional framework reflects aspects of change, especially the establishment of the home and com-munity-based services archetype, it also displays inertia including the continued dominance of the nursing home archetype. Roles played in these contested dynamics are traced along a key process of change in each archetype: (1) the growth of large multi-facility (chain) nursing home corporations, and (2) political advocacy for home and community-based services. PMID- 15779469 TI - A theory of physician-hospital integration: contending institutional and market logics in the health care field. AB - This article proposes a theory of physician-hospital integration. The theory is developed by building on three streams of scholarship: "new" institutionalism, "old" institutionalism, and the theory of economic markets. The theory uses several key concepts from these theoretical frameworks, including the notions of environmental demands for legitimacy, market demands for efficiency, and agency. To enhance the predictive power of the theory, two new concepts are introduced: directionality of influence between institutional and market forces at the macro societal level, and degree of separation of institutional and market domains at the local level--which add important predictive power to the theory. Using these concepts, a number of hypotheses are proposed regarding the ideal types of physician-hospital arrangements that are likely to emerge under different combinations of directionality of influence and institutional and market domain separation. Moreover, the theory generates hypotheses regarding organizational dynamics associated with physician-hospital integration, including the conditions associated with high and low prevalence of physician-hospital integration, the extent to which the integrated organization is physician-centric or hospital centric, and whether physician-hospital integration is likely to be based on loose contractual arrangements or tight, ownership-based arrangements. PMID- 15779470 TI - Community responses to national healthcare firms. AB - Over the last 25 years, national Health Maintenance Organization (HMO) and hospital firms attempted to enter local markets, either by acquiring formerly independent, locally based HMOs and hospitals or by directly entering local markets. While national HMOs have been relatively successful, national hospital firms have had much less success. This paper explores the reasons for this difference. It reviews changes in presence of national HMO and hospital firms in markets, discusses common conceptual lenses through which national entry into local markets typically has been viewed, and shows how social network theory can be used to develop a better understanding of why the entry experience of national HMO and hospital firms varies across markets. The paper concludes with a research agenda that addresses issues raised by social network theory and its application to national firm entry into local markets. PMID- 15779472 TI - Medicalization, markets and consumers. AB - This paper examines the impact of changes in the medical marketplace on medicalization in U.S. society. Using four cases (Viagra, Paxil, human growth hormone and in vitro fertilization), we focus on two aspects of the changing medical marketplace: the role of direct-to-consumer advertising of prescription drugs and the emergence of private medical markets. We demonstrate how consumers and pharmaceutical corporations contribute to medicalization, with physicians, insurance coverage, and changes in regulatory practices playing facilitating roles. In some cases, insurers attempt to counteract medicalization by restricting access. We distinguish mediated and private medical markets, each characterized by differing relationships with corporations, insurers, consumers, and physicians. In the changing medical environment, with medical markets as intervening factors, corporations and insurers are becoming more significant determinants in the medicalization process. PMID- 15779473 TI - Evidence-based medicine and the reconfiguration of medical knowledge. AB - Over the past decade, different parties in the health care field have developed and disseminated clinical practice guidelines as part of evidence-based medicine. These formal tools based on a scientific evaluation of the research literature purport to tell health care professionals how to practice medicine. Because clinical practice guidelines shift the knowledge base in the health care field through standardization, they remain controversial within and outside medicine. In this paper, we evaluate the predictive accuracy of four medical professionalization theories--functionalism, Freidson's theory of professional dominance, deprofessionalization theory, and the theory of countervailing powers- to account for (1) the shift from pathophysiology to epidemiology with guidelines, (2) the creation of practice guidelines, and (3) the effects of clinical practice guidelines on the autonomy of health professionals. In light of the mixed predictive record of professionalization theories, we conclude with a need for "evidence-based sociology" and a recalibration of basic premises underlying professionalization theories. PMID- 15779471 TI - Multinational corporations and health care in the United States and Latin America: strategies, actions, and effects. AB - In this article we analyze the corporate dominance of health care in the United States and the dynamics that have motivated the international expansion of multinational health care corporations, especially to Latin America. We identify the strategies, actions, and effects of multinational corporations in health care delivery and public health policies. Our methods have included systematic bibliographical research and in-depth interviews in the United States, Mexico, and Brazil. Influenced by public policy makers in the United States, such organizations as the World Bank, International Monetary Fund, and World Trade Organization have advocated policies that encourage reduction and privatization of health care and public health services previously provided in the public sector. Multinational managed care organizations have entered managed care markets in several Latin American countries at the same time as they were withdrawing from managed care activities in Medicaid and Medicare within the United States. Corporate strategies have culminated in a marked expansion of corporations' access to social security and related public sector funds for the support of privatized health services. International financial institutions and multinational corporations have influenced reforms that, while favorable to corporate interests, have worsened access to needed services and have strained the remaining public sector institutions. A theoretical approach to these problems emphasizes the falling rate of profit as an economic motivation of corporate actions, silent reform, and the subordination of polity to economy. Praxis to address these problems involves opposition to policies that enhance corporate interests while reducing public sector services, as well as alternative models that emphasize a strengthened public sector PMID- 15779475 TI - Legislation and our profession. PMID- 15779474 TI - Global manpower needs for integrative systems physiologists. PMID- 15779476 TI - Volunteerism: limitless opportunities. PMID- 15779477 TI - Now more than ever, develop professionally through local connections. PMID- 15779478 TI - Modern electric handpieces feature improved benefits for today's dental surgeon. PMID- 15779479 TI - Laser safety officer: a vital role for dental assistants. AB - In closing, the LSO is a vital role for dental assistants and one that will expand as laser use increases. The Academy of Laser Dentistry (ALD) is your source and resource for information and certification in order to garner this status. Together with the ALD, dental assistants will have the knowledge and skills to implement a successful laser program within individual practices. The members of your profession must not lose this opportunity to further their education. Laser dentistry is gaining acceptance because of the benefits not only to the practice but to patients. Dental assistants can and should provide this vital role in the safe and effective delivery of this technology so that the best care is delivered. PMID- 15779480 TI - Intraarterial chemotherapy: another choice for unresectable advanced hepatocellular carcinoma? PMID- 15779481 TI - The metabolic syndrome: what have we learned? PMID- 15779482 TI - Benefits of surgery for patients with pulmomary metastases from hepatocellular carcinoma. PMID- 15779483 TI - Clinical effects of intra-arterial infusion chemotherapy with cisplatin, mitomycin C, leucovorin and 5-flourouracil for unresectable advanced hepatocellular carcinoma. AB - BACKGROUND: Intra-arterial infusion chemotherapy (IAIC) can potentially improve survival in some patients with hepatocellular carcinoma (HCC), but the ideal regimen is not yet established. We prospectively evaluated the effects of short course continuous infusion with the combination of cisplatin, mitomycin C, 5 fluorouracil (5-FU) and leucovorin for unresectable advanced HCC and analyzed their prognostic factors. METHODS: Patients with unresectable advanced HCC and not suitable for other therapy were enrolled. Cannulation via the left subclavian artery with the tip of catheter at the proper hepatic artery was done before initialization of IAIC routinely. The regimen consisted of the daily administration of cisplatin (10 mg/m2), mitomycin C (2 mg/m2), leucovorin (15 mg/m2), and daily infusion of 5-FU (100 mg/m2) for 5 days. Only the patients that had received at least 2 courses of IAIC were evaluated. RESULTS: Two-hundred and 11 courses of IAIC were performed, and each patient received at least 2 cycles of chemotherapy. The overall response rate was 28.3%. We observed a complete response in 5 patients (9.4%), a partial response in 10 patients (18.9%), a minimal response in 5 patients (9.4%), no change in 11 patients (20.8%) and a progressive disease in 22 patients (41.5%). The patients with response to treatment survived longer than the patients without response (24.6 +/- 14.2 months vs 8.7 +/- 5.3 months, p < 0.001). In univariate and multivariate analysis, absence of main vessel thrombosis and alpha-fetoprotein (AFP) reduction percentage > 50% following treatment showed significance in our study. All side effects subsided after conservative treatment. CONCLUSIONS: Continuous IAIC with cisplatin, mitomycin-C, leucovorin, and 5-FU is effective for patients with severe advanced HCC. Absence of main vessel thrombosis, and AFP reduction percentage > 50% following treatment were good predictors of treatment response in our study. All side effects were mild and tolerable. PMID- 15779484 TI - Prevalence of metabolic syndrome in a large health check-up population in Taiwan. AB - BACKGROUND: Metabolic syndrome is a major risk factor for cardiovascular disease and the development of type 2 diabetes. The purpose of the present study was to investigate the prevalence of metabolic syndrome in a large health check-up population in Taiwan. METHODS: During the period of 2000-2001, about 120,000 adults aged 20 years and above received health check-ups at the MJ Health Screening Centers. Among these, 24,329 participants with complete data on waist circumference, blood pressure, fasting levels of glucose, triglyceride, and high density-lipoprotein cholesterol were randomly selected for analysis. The prevalence of metabolic syndrome was estimated according to the criteria proposed by the National Cholesterol Education Program (NCEP III), with adoption of the Asian criteria for abdominal obesity. RESULTS: Based on the strict NCEP III criteria, the prevalence of metabolic syndrome was 9.5% (10.6% in men and 8.1% in women, respectively); with Asian criteria for waist circumference, 12.9% (15.5% in men and 10.5% in women, respectively). The prevalence of metabolic syndrome increased with age. Prevalences of all components of metabolic syndrome increased sharply with age in women. In contrast, the prevalences of high triglyceride and low high-density-lipoprotein cholesterol flattened and declined after the age of 50 years in men. Therefore, the gender difference resulted in a cross-over of the prevalence of metabolic syndrome between men and women at the age of 60 years. CONCLUSIONS: The prevalence of metabolic syndrome in a physical check-up population is significantly affected by age and gender. Age- and gender-specific strategies may be useful in the control of the syndrome. PMID- 15779485 TI - Pulmonary metastasectomy for hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is seldom resectable due to advanced status. Even though hepatectomy is feasible, a large proportional of patients may still develops extrahepatic recurrence. Pulmonary metastasis is the most common site of extrahepatic spread. Few articles have discussed the benefit of resection for lung metastasis after curative hepatectomy. We evaluated the general information and the result of lung resection for patients having lung metastasis after curative resection of HCC. METHODS: Six patients who underwent pulmonary metastasectomy for HCC at Taipei Veterans General Hospital between August 1995 and May 2004 were enrolled in the study. All of them had received the hepatectomy for primary HCC. The demographic information of patients, the site and number of extrahepatic recurrence, the method of surgical intervention and the outcome after surgery were retrospectively reviewed. RESULTS: There were 4 men and 2 women with the mean age of 47.3 years. All of them were HBV carriers. Five patients had multiple pulmonary metastases removed by wedge resection. Two patients had bilateral lung metastases upon diagnosis of extrahepatic recurrence. The mean duration of follow-up after hepatic resection was 75.0 +/- 25.1 months (32 to 104 months). The mean survival after pulmonary resection was 47.2 +/- 34.3months (1 to 94 months). Four patients are still alive and free of the disease. One patient is alive but with the disease. One patient who refused further aggressive treatment after resection of lung metastasis died of the disease 40 months after lung resection, 77 months after hepatic resection. CONCLUSIONS: Lung resection for the pulmonary metastasis of HCC can result in a favorable long-term survival when there is no other intrahepatic or extraphepatic recurrence of HCC. For patients with multiple lung metastases in different lobes or different lungs, aggressive surgical resection is recommended if complete resection can be achieved. PMID- 15779486 TI - Urolithiasis in pregnancy. AB - BACKGROUND: The aim of this study was to review our experience in the diagnosis and management of urolithiasis in pregnant women, and to discuss safety, appropriate diagnostic tools and treatment modalities with review of literatures. METHODS: Nine women with pregnancy proved to have urolithiasis were included in this study. The diagnosis was made according to their symptoms and signs, and ultrasonographic findings. Conservative treatment with hydration, analgesics and antibiotics was applied initially, followed by some minimally invasive procedures, including percutaneous nephrostomy (PCN), double J (DBJ) stenting and ureterorenoscopy (URS), if initial treatment failed. RESULTS: All patients received conservative treatment at first, and 4 patients got well till delivery, another 5 patients needed further minimally invasive procedures; 1 of them received DBJ stenting, another 1 received PCN first and changed to DBJ stenting. The remaining 3 patients were treated by ureterorenoscopy with stone manipulation. All these 5 patiens had full-term delivery without obstetric sequela, except 1 patient who received left salpingoophrectomy due to left ovarian abscess. CONCLUSIONS: During pregnancy, urolithiasis is a diagnostic and therapeutic challenge. Clinical symptoms and signs are most important in establishing diagnosis, and ultrasound examination adds accuracy. Minimally invasive procedures (such as PCN, DBJ stenting, URS) can facilitate the successful management of patients requiring further intervention due to failure of conservative treatment. PMID- 15779487 TI - Primary appendiceal lymphoma presenting as perforated acute appendicitis. AB - Acute perforated appendicitis is a very rare initial presentation of appendiceal lymphoma. A case of primary lymphoma of the appendix in a 42-year-old female is reported here. The symptom was pain in the right lower quadrant of the abdomen. Surgical intervention was performed under clinical diagnosis of acute appendicitis. Grossly, the resected appendix was gangrenous and perforated. Light microscopy revealed transmural infiltration by diffuse large B-cell lymphoma with angioinvasion and tumor necrosis, resulting in perforated acute suppurative appendicitis. To the best of our knowledge, only 4 cases of well-documented primary diffuse large B-cell lymphoma of appendix have been reported in the world literature. PMID- 15779488 TI - Pilomatrixoma of the head and neck. AB - Pilomatrixoma is a benign skin appendage tumor that commonly occurs as a solitary lesion and is not usually hereditary. There is evidence to suggest that patients with a family history of multiple pilomatrixomas have a high probability of autosomal dominant disorders such as myotonic dystrophy, Gardner syndrome, and Rubinstein-Taybi syndrome. In January 2004, a case was reported of an 8-year-old girl with 2 progressively enlarged facial masses. One of them was excised and diagnosed as pilomatrixoma. Coincidently, her 13-year-old sister had the same type of tumor when she was 4 years old. We report this unusual case and review the literature. Pilomatrixoma has not been widely reported in the head and neck surgery literature. This benign tumor may be misdiagnosed as a carcinoma, resulting in unnecessary aggressive therapy. Otolaryngologists should therefore note the clinical and pathologic characteristics of these symptoms. PMID- 15779489 TI - Extraperitoneal rectal perforation without perineal wound or pelvic fracture. AB - The present report describes an unusual case of rectal perforation. An 81-year old female was struck by a truck while walking in the street, and she was sent to a local hospital where fracture of the right subtrochanteric femur was diagnosed. She was admitted, in stable condition, for planned orthopedic operation. Consciousness change and respiratory distress developed 6 hours later. She was then transferred to a trauma center where extraperitoneal rectal perforation was diagnosed. Despite empirical antibiotics and surgical intervention, the patient unfortunately expired 3 days later. Unusual mechanism and incomplete physical examination were the major causes of delayed diagnosis. This case report also discusses the mechanism, classification and management of rectal perforation. PMID- 15779490 TI - Subarachnoid hemorrhage and death following coingestion of MDMA with other drugs. AB - Ecstasy, the popular name for 3,4-methylenedioxymethamphetamine (MDMA), is a synthetic amphetamine derivative. It stimulates the sympathetic nervous system, producing serious adverse effects on the cardiovascular system. We present a 20 year-old female patient, who developed subarachnoid hemorrhage (SAH) and death following MDMA and coingestion with other drugs. She suffered from severe headache followed by vomiting, and conscious change 5 hours after an intake of 1 tablet MDMA and other drugs at a dance club. Her blood pressure was 226/164 mmHg, pulse rate 164/min, respiratory rate 30/min on arrival at our emergency department. Diffuse rales were heard over both lung fields. Both pupils' sizes were 4 mm, with sluggish reaction to light. A 12 lead electrocardiograph showed sinus tachycardia, ST depression in the inferior leads and V4 to V6 precordial leads. Laboratory findings revealed normal except a slightly raised white cell count and glucose. Arterial blood gas analysis showed pH was 7.333, with PaCO2 24.6 mmHg, PaO2 151.7 mmHg and HCO3 12.8 mmol/L. Chest x-ray revealed acute pulmonary edema. Urgent computerized tomography scanning of the head demonstrated SAH. Her condition continued to deteriorate, and went to deep coma and shock status. She expired on the second day although we treated aggressively. PMID- 15779491 TI - Femoral neuropathy after pelvic surgery. AB - Postoperative femoral neuropathy is an uncommon complication occurring after pelvic surgery. Inappropriate stretching and prolonged compression of the nerve are 2 major mechanisms of the neuropathy. Here we report 2 cases of femoral neuropathy immediately following pelvic surgery. Both cases had neither previous vascular nor peripheral nerve disease. They suffered from weakness of left hip flexion and knee extension and sensory impairment over the left lower limb after surgery. Electromyography and nerve conduction studies confirmed left femoral neuropathy. Both of the patients received physical therapy and had nearly total neurological recovery within 3 months. We report this unusual complication that followed major pelvic surgery and also review the literature and discuss the possible etiology for prevention of this injury. PMID- 15779492 TI - A mandibular advancement device reduces sleep disordered breathing in patients with congestive heart failure. AB - Sleep disordered breathing (SDB) including obstructive and central sleep apnoea/hypopnoea as well as periodic breathing (PB) is common and is believed to increase risk for mortality in patients with congestive heart failure (CHF). Mandibular advancement device (MAD) has widely been recommended for treatment of obstructive sleep apnoea but the method has never been investigated for treatment of SDB in the patients with CHF. The aim with the present study was to examine the effect of MAD intervention on SDB in patients with CHF. The study included 17 male patients, aged 68.4+/-5.7 (mean+/-SD) with stable, mild to moderate CHF due to left ventricular systolic dysfunction and with SDB, expressed as apnoea/hypopnoea index (AHI) > or = 10. The SDB was examined during a single night using an unattended, portable polysomnographic device in the patients home, prior to and following intervention with a individually adjusted MAD. The SDB was evaluated by calculating AHI, PB expressed as the percentage of the total registration time, oxygen desaturation index (ODI) and snoring time. The AHI was reduced by MAD intervention from 25.1+/-9.4 to 14.7+/-9.7 (p=0.003). ODI reduced from 21.1+/-9.0 to 10.5+/-7.8 (p=0.007) and snoring time decreased from 53+/-111 to 18+/-47 seconds (p=0.02). PB was reduced from 55.7+/-25.6 to 40.4+/-26.4 per cent without statistical significance. In conclusion, the MAD intervention may be a feasible method for reducing SDB in patients with stable, mild to moderate CHF and left ventricular systolic dysfunction. PMID- 15779493 TI - Knowledge on periodontal disease before and after a mass media campaign. AB - The aim of this study was to evaluate if a mass media campaign regarding periodontal disease could increase the knowledge in the general population of diagnoses, symptoms and treatment options of periodontal disease. More specifically, the aim was to investigate the number of correct answers to knowledge questions before and after the campaign. The Swedish Association of Periodontology conducted the campaign through brochures, newspapers, radio and TV. The effect of the campaign was evaluated by a pre- and post campaign questionnaire with a cohort design. From a national population register of 50-75 year olds in Sweden, 900 persons were randomly sampled for the study. A total of 64% of the sample answered both questionnaires. The result of the study showed an improvement among the respondents. There was a significant increase in the number of correct answers regarding diagnoses, symptoms and treatments of periodontitis. In the questionnaire, correct answers regarding "Mobile teeth" increased from 57% to 65% (p=0.003) and "careful dental hygiene" from 65% to 73% (p=0.001). Kappa value's were calculated for consistency in the reply and all kappa values were low especially for the questions "X-ray" (0.36) and "Cleaning between the teeth" (0.38). It was concluded that the campaign probably was successful from a public health knowledge standpoint. PMID- 15779494 TI - In vivo validity and reliability of IR fluorescence measurements for caries detection and quantification. AB - Studies of detection and quantification of dental caries by KaVo DIAGNOdent have shown high diagnostic accuracy and high inter-operator agreement in vitro. The aim of the present study was to determine whether comparable results could be achieved under clinical conditions. In Part I (occlusal surfaces), 52 occlusal sites (suspected caries) were examined by visual inspection, bitewing radiography, and two DIAGNOdent devices. The fissures were then opened, and lesion depth established according to a four point scale. In Part II (smooth surfaces), two operators performed DIAGNOdent measurements of incipient carious lesions on the buccal surface of 30 molar teeth. The readings were compared with corresponding measurements by Quantitative Light-induced Fluorescence (deltaF). The results for occlusal surfaces disclosed very low correlation between lesion depth and readings by the two DIAGNOdent devices. The intra-operator agreements were very good (0.80-0.92), and the inter-operator agreement was good (0.71 0.87). A significant systematic difference in readings by the two devices precludes general recommendations of cut-off levels. However, individually suitable cut-off values for dentinal lesions gave good qualitative information about the surface (77%-85% over all correct observations), rather than quantitative information in terms of lesion depth within the dentine. In this context, the DIAGNOdent device might be of help for the clinician in the process of decision making about invasive therapy. For smooth surfaces, the correlation between readings obtained with the DIAGNOdent device and deltaF were acceptable (0.57-0.73). The intra-operator agreement was 0.94 for both operators (excellent), and the inter-operator agreement ranged from 0.79 to 0.87 (very good). PMID- 15779495 TI - Racial and ethnic disparities in cardiac catheterization for acute myocardial infarction in the United States, 1995--2001. AB - OBJECTIVE: To examine recent trends in racial and ethnic disparities in cardiac catheterization for acute myocardial infarction (AMI) to determine whether disparities documented from the 1980s through mid-1990s persist, and evaluate whether patient and hospital characteristics are associated with any observed disparities METHODS: Cross-sectional analyses of 585,710 white, 51,369 black and 31,923 Hispanic discharges from hospitals in the Nationwide Inpatient Sample (which includes data on all discharges from 951 representative hospitals in 23 states) that had performed cardiac catheterization from 1995--2001 with a primary diagnosis of AMI. Adjusted procedure rates and prevalence ratios (PR) were computed to compare catheterization rates by race and ethnicity. MEASUREMENTS AND MAIN RESULTS: Catheterization rates were higher for whites than blacks for all years examined; rates among Hispanics increased during this period and approached the rate among whites. After adjustment for age, demographics, comorbidity, year and hospital characteristics, rates (per 100 discharges) were 58.4 for whites, 50.1 for blacks (PR 0.87; 95% CI 0.84-0.91) and 55.2 for Hispanics (PR 0.95; 95% CI 0.90-0.99). CONCLUSIONS: These nationwide data suggest blacks remain less likely than whites and Hispanics to undergo catheterization during a hospitalization for AMI. Whether this disparity stems from patient or provider factors remains to be determined. PMID- 15779497 TI - Family poverty accounts for differences in lower-extremity amputation rates of minorities 50 years old or more with diabetes. AB - Rates of leg amputations in diabetics are known to differ among racial/ethnic groups, but the relationship between family poverty and the risk of amputation has not been fully addressed. One-hundred-seven diabetic patients, all 50 or more years old and all from ZIP code tabulation area 778, underwent their first amputations at one hospital. Linear regression evaluated differences in age and atherosclerosis severity among African-American, Hispanic-American and other patients. chi2 statistics evaluated differences among African-American, Hispanic American and other patients, with respect to sex and type of amputation. chi2 statistics evaluated differences among fractions of African- American, Hispanic American and other patients, with respect to those residents 50 years old and older and those of poor families. Patient groups did not differ in regard to age, atherosclerosis severity, sex or type of amputation (P>0.05). The percents who were African-American, Hispanic-American, and other (33%, 21%, and 47%, respectively) differed markedly from those of persons 50 or more years old [13%, 7%, and 79%, (w=0.81, P<0.00001)] and mirrored those of poor families [37%, 19%, and 44% (w=0.08, P>0.05)]. Family poverty accounts for differences in diabetic amputation rates of African Americans, Hispanic Americans and other persons 50 or more years old. PMID- 15779496 TI - Relationship of C-reactive protein and bone mineral density in community-dwelling elderly females. AB - OBJECTIVE: Inflammatory cytokines have been shown to play an important role in bone remodeling. We hypothesized that higher levels of C-reactive protein (CRP) are associated with low bone mineral density (BMD) in elderly females. DESIGN: Secondary data analysis of the Third National Health and Nutrition Examination Survey. PARTICIPANTS: 2,807 females 65 years and older. RESULTS: CRP was associated with BMD in the bivariate sis (p<0.001) but not in the multivariate analysis (p=0.23) Age, ethnicity, body mass index (BMI), hormone replacement therapy (HRT) and immobility were independently associated with BMD. CONCLUSIONS: CRP may be useful in screening for osteoporosis among community-dwelling elderly females. However, CRP appears to act as a surrogate for other factorsdirectly associated with osteoporosis. Further studies are needed to validate these findings. PMID- 15779499 TI - Nonductal dependent coarctation: a 20-year study of morbidity and mortality comparing early-to-late surgical repair. AB - OBJECTIVE: The objective of this study was to evaluate the effect of the timing of surgical repair of nonductal dependent coarctation on the short-term outcome. METHODS: The medical records of 76 patients, diagnosed and treated for a nonductal dependent mild-to-moderate coarctation at a tertiary care institute over a 20-year period, were retrospectively reviewed with the age at repair compared against outcome measures. Multiple logistic regression was performed to assess the timing of repair, the presence of congestive heart failure or associated cardiac defects on the outcome measures. RESULTS: The mean age of surgery for the mild-to-moderate coarctation repair was 3.1 years (range: three days to 12 years). The most common cause for referral to a pediatric cardiologist was the clinical finding of a cardiac murmur. The timing of surgical repair was not found to be a predictor of morbidity or mortality. There was no significant difference in outcome measures defined as residual hypertension, residual coarctation gradient, persistent cardiomegaly, postoperative neurological sequelae, the requirement for a second surgery or the need for balloon dilatation for residual postoperative coarctation and the need for antihypertensive medications within five years postsurgery. CONCLUSION: The timing of surgical repair in the setting of nonductal dependent, mild-to-moderate coarctation of the aorta, does not adversely affect the short term (less than 20 years) outcome in children. PMID- 15779498 TI - Moving beyond attitudinal barriers: understanding African Americans' support for organ and tissue donation. AB - PURPOSE: African Americans are disproportionately represented among individuals in need of an organ transplant, due in part to low donation rates in this population. The research literature has focused on attitudinal barriers to donation; however, the current study explores individual experiences and values that contribute to supportive attitudes toward organ and tissue donation. PROCEDURES: Focus group participants were 26 African-American clergymen and 42 African-American parishioners recruited from seven Christian churches in the metro Atlanta area. FINDINGS: Although a large number of participants had previous exposure to organ and tissue donation and transplantation, the majority of these experiences were negative, and participants felt a general fear and lack of knowledge about the process of donation and transplantation. Despite these negative experiences, respondents reported personal values (e.g., the desire to help others and acceptance of group responsibility) and religious values (e.g., the desire to carry out God's will and to have faith in God) that contribute to supportive attitudes toward organ and tissue donation. CONCLUSION: An understanding of supportive attitudes toward donation may help improve the development of effective culturally sensitive intervention messages targeting the African-American religious community with the ultimate goal of increasing the pool of organs available for transplantation. PMID- 15779500 TI - Minority status and the risk of serious childhood injury and death. AB - OBJECTIVE: Minority populations have an increased risk for trauma, but little is known about injury rates for minority children. This study compares the causes, rates and outcomes of traumatic injuries between minority and white children in a statewide population sample. METHODS: A cohort study of 5,973 children (age <16) receiving inpatient care for treatment of acute injuries at the pediatric trauma centers in Ohio from 1999--2001. Case records were analyzed for race, injury type, injury severity, length of stay and demographic information. Supplemental data sources included the 2000 U.S. census and Ohio Vital Statistics death certificates 1996--2001. MAIN OUTCOME MEASURES: Hospital admission rate, mortality rate, length of stay, rate of admission to rehabilitation service. RESULTS: African-American children, who composed the vast majority of the minority population sample, were 7.7 more times likely to sustain a burn or gunshot wound, seven times more likely to be struck by a car, six times more likely to be intentionally injured and over twice as likely to killed by an injury than white children. However, after adjusting for injury severity, they have the same mortality, hospital length of stay and referral rate to rehabilitation as white children. CONCLUSION: Trauma has a far greater impact on minority children than on white children. Research and development of injury prevention initiatives that specifically target minority children are urgently needed. PMID- 15779501 TI - Obstructive sleep apnea syndrome in a publicly funded healthcare system. AB - BACKGROUND: Despite its current recognition as a major health concern, little has been published about obstructive sleep apnea syndrome (OSAS) as a health problem in public healthcare systems where limited resources, language and cultural differences may present barriers to detection and treatment. OBJECTIVE: To describe patients referred for suspected OSAS in a large county-funded healthcare system. METHOD: A retrospective, descriptive observational study that included all patients referred for an OSAS evaluation between September 2000 and September 2002. RESULTS: Only 123 patients were referred and 115 completed an evaluation during the two-year period: 99% met OSAS diagnostic criteria, which was severe in 79% and frequently complicated by related comorbid conditions. CPAP acceptance was lower than in the previous series, especially among Hispanics. CONCLUSIONS: The findings suggest that referral for OSAS evaluation was limited to those most severely affected and raise the possibility of underdetection and undertreatment in the ublic sector. PMID- 15779502 TI - Chronotherapeutic oral drug absorption system verapamil is effective in reducing morning blood pressure in African Americans: a post hoc analysis of the chrono trial. AB - Results of several clinical trials have shown that verapamil is effective in reducing blood pressure (BP) in African Americans, a population at high risk for hypertension and target-organ damage. Nonetheless, adequate control of BP is perceived as difficult to achieve in this population. A post hoc analysis of data from the community-based CHRONO trial (Controlling Hypertension in the moRning with a ChrONO medication) was undertaken to assess racial/ethnic differences in the safety and efficacy of the Chronotherapeutic Oral Drug Absorption System (CODAS) formulation of verapamil in a real-world setting. Once-daily administration of the CODAS formulation of verapamil significantly reduced morning BP (P<0.0001) regardless of race or ethnicity. In the African-American population (N=466), the response rate for systolic BP (<140 mmHg or > or =10% reduction from baseline) and diastolic BP (<90 mmHg or reduction > or =10 mmHg from baseline) combined was 70.8%, and 60% of those individuals responded at the lowest (200 mg) dose. Of the 59.7% of African Americans who reached the target BP of <140/90 mmHg, 64% did so at the 200-mg dose. Response rates were not affected by gender, age or treatment history, and CODAS-verapamil was well tolerated in all ethnic/racial treatment groups. In a trial conducted in actual clinical practices, the CODAS formulation of verapamil was shown to be safe and effective in African Americans, Caucasians, Hispanics and Asians. PMID- 15779503 TI - Clinical characteristics and echocardiographic features of black men vs. black women with systolic heart failure. AB - PURPOSE: We sought to determine whether significant differences exist in clinical characteristics and echocardiographic features between black men and black women who presented to an outpatient heart failure clinic. METHODS: A retrospective review of the charts of 114 consecutive patients with systolic dysfunction who presented to a heart failure clinic in Brooklyn from 1999--2003 was performed. The first echocardiogram within six months of presentation to the clinic was used for analysis. The clinical characteristics included are: age, sex, body mass index (BMI) and diabetes status. A significance level of <0.05 was used throughout. SUMMARY: Based on the analysis of 108 black men and women, overall, men have a significantly larger left ventricular end-diastolic diameter (LVEDD) compared to women (p<0.006). Younger men (7.17 cm) had a larger LVEDD compared to older men (6.37 cm) and both younger and older women (6.22 cm and 6.40 cm, respectively). This difference in LVEDD between the sexes decreased with aging. Men (22.17) had a lower ejection fraction (EF) than women (25.37). No significant differences were noted in the BMI (30.45 for men and 28.60 for women) and the albumin level (3.59 for men and 3.44 for women) between the sexes. However, the younger age group (particularly younger men) had a higher BMI than the older age group (p=0.008). PMID- 15779504 TI - Geriatric autopsy findings in the last 10 years: an Urban Teaching Hospital experience. AB - In this study, we reviewed autopsy records for clinical data and autopsy findings from patients aged 70 or more, over a 10-year period (1993 to 2002) in an urban university hospital. For that period, there were a total of 772 autopsy cases of which 180 (23%) patients were aged 70 years or older. We found that despite a marked decrease in total autopsy rates, there has been a perceptible rise in geriatric cases. Cardiovascular and infectious diseases in this age group are the leading causes of death as reported nationally. We found that women died more of acute myocardial infarctions than men, even though hypertensive and atherosclerotic cardiovascular diseases not otherwise specified were more prevalent in men. It is our conclusion that at our institution: 1) despite a marked decrease in the total autopsy rate, the geriatric autopsy rate is rising; 2) infectious and cardiovascular diseases are the leading causes of death in elderly patients; 3) Myocardial infarcts as a cause of death are more often seen in women for this age group. It is also our impression that better autopsy reporting is needed for maximal utilization of autopsy findings in medical auditing and teaching and for improvements in the quality of patient care in general and the geriatric patient in particular. PMID- 15779505 TI - Diagnosis of tuberculosis: urgent need to strengthen laboratory services. AB - Tuberculosis (TB) is of great public health concern globally, and the impact is most felt in developing countries of Asia and Africa, where 95% of cases and 98% of deaths are attributable to the disease, The disease is poverty-driven and the situation is further worsened by absence of rapid diagnostic tools to facilitate early diagnosis of the illness, thus leading to widespread of the disease. While attention is being focused on the HIV/AIDS pandemic, little is being heard of TB, especially in the areas of laboratory diagnosis despite the fact that the disease is the commonest cause of death in people living with HIV/AIDS. The importance of a diagnostic laboratory in a TB control program cannot be overemphasized. Smear microscopy, which is the cornerstone of World Health Organization (WHO) 'DOTS' strategy for the treatment of TB, has many drawbacks among which is its inability to detect latent infection and the dependency of its sensitivity on a trained and motivated microscopist. Therefore, there is a need for a more reliable, sensitive and rapid diagnostic test to facilitate early diagnosis of cases and prompt initiation of therapy for a TB control program to have a meaningful impact in the community. PMID- 15779506 TI - Medical care on the brink: the need for re-engineering healthcare services in sub Saharan Africa. AB - The sub-Saharan region of Africa is home to more of the ills of mankind than any other region on earth. Nowhere is the aggregate of disease, political turmoil, inadequate resources and a crumbling infrastructure so completely packaged in a seemingly "escape proof" pod as in sub-Saharan Africa. This continent is a kaleidoscope of people and problems derived from artificial boundaries drawn by European colonial powers, resulting in a litany of problems that have flourished for many decades. In the immediate postcolonial era, there was some oversight by the departed powers, but this has changed recently with decreasing interest in African affairs and only episodic worldwide news coverage because of other world events that overshadow Africa and its problems. The end of the cold war also eliminated the attention Africa received when the superpowers were courting nations. The American Medical Team for Africa has conducted medical missionary work throughout Africa for over a decade and, through its observations, has developed recommendations that are germane to all of sub-Saharan Africa. The organization thinks that this might warrant the attention of governments, international pharmaceutical houses, foundations, the United Nations and all international aid agencies concerned about the plight of healthcare in Africa. These recommendations should enable these countries to re-establish an affordable, efficient and sustainable infrastructure for basic hospital services so that they can diagnosis, monitor, treat and manage disease populations. In some areas, Africa needs to be retrofitted with technology from the past, while in others it needs to be fast-forwarded into the future. The purpose of this manuscript is to try putting the various healthcare challenges into one of these two categories. PMID- 15779507 TI - A comparative study of academic achievement of children with sickle cell anemia and their healthy siblings. AB - BACKGROUND: Sickle cell anemia (SCA) could impact negatively on academic performance of affected children. Reports so far have been inconsistent. OBJECTIVE: To do a comparative analysis of academic performance of Nigerian children with SCA and their siblings. METHODS: We studied sessional aggregate scores, pass rates and percentage scores in four core subjects (mathematics, English language, integrated science and social studies), as well as the total number of days of school absence of 52 school-age children (6-17 years) with sickle cell anemia and 42 siblings of similar sociodemographic characteristics, over one academic session. FINDINGS: School absence among the SCA patients was significantly higher than that of siblings (9.3+/-5.5 days/school year and 4.3+/ 2.6 days/school year, respectively, mean+/-SD, p<0.05). Although the mean sessional aggregate score for patients was comparable with that of the siblings, there was a significantly larger proportion of below-average pupils among the patients. The mean percentage scores of the patients on mathematics, English language, integrated science and social studies compared well with those of the siblings. There was no significant correlation between school absence and academic achievement of the study population. CONCLUSION: More children with SCA are underachievers in this study, and their school underachievement is not associated with the observed higher school absence. PMID- 15779508 TI - What does the geometry of an arc reveal about methods of assessing spinal radiographs in scoliosis? AB - BACKGROUND: The shapes of an arc and scoliosis are the same. Therefore, the geometry of an arc can reveal the relationship between the angles within and around a scoliotic spine. OBJECTIVE: To look for angles which are functions of the geometry of an arc and can also be used for assessment of curve progression in scoliosis. DESIGN: Review of the geometry of an arc and that of scoliosis. METHOD: English literature on scoliosis (from 1930 to date), searched manually, via Internet and from colleagues, was reviewed. RESULTS: Methods of curve measurement in use presently are functions of the geometry of scoliosis. Three additional methods are proposed for assessment of curve progression in scoliosis. These methods are geometrically related with the previously known ones. CONCLUSION: All methods of assessment of curve progression in scoliosis are fundamentally the same and interchangeable. There is no reason for preference of one method over another method of curve measurement. PMID- 15779509 TI - Acute kidney transplant failure following transurethral bladder polyp fulguration. AB - Ureteral obstruction and anastomotic leak represent the most common urologic complications of kidney transplantation. Delay in diagnosis or treatment can lead to allograft loss. Obstruction of the ureter occurs in 2% of kidney transplant recipients. Although the majority of cases are immediate technical complications of the operation, subsequent manipulation of the genitourinary system can result in iatrogenic ureteral injury. We report the case of a long-term kidney transplant recipient who developed obstructive uropathy and acute renal failure requiring dialysis after undergoing cystoscopy and bladder polyp fulguration. The etiology was inadvertent thermal injury of the ureteroneocystostomy incurred during the procedure. After attempted percutaneous management, definitive open repair resulted in a return of allograft function to baseline. PMID- 15779510 TI - Transfusion-associated graft-versus-host disease in an immunocompetent individual following cardiac surgery. AB - A 65-year-old man underwent coronary artery bypass graft surgery at our tertiary care hospital. Perioperatively, he was transfused with four units of nonirradiated whole blood from first-degree relatives and discharged from the hospital at postoperative day seven. He presented six days later with fever, skin rash, elevated liver enzymes, and progressive pancytopenia. Elevated bilirubin levels and diarrhea were added to the clinical picture over the following days. Clinical findings and results of a skin biopsy specimen were consistent with transfusion-associated graft-versus-host disease. The patient died 20 days after transfusion. PMID- 15779512 TI - Maximizing reimbursement by appealing claim denials. AB - Providers continue to lose millions of dollars from payers' abusive practices. An immediate way to increase reimbursement rates is to know and understand effective administrative appeal processes. These processes are usually defined by contract or by statute. Recent amendments to regulations governing ERISA are helpful to providers. PMID- 15779511 TI - A case of systemic malignant atrophic papulosis (Kohlmeier-Degos' disease). AB - Malignant atrophic papulosis (MAP) is a rare and clinically distinctive vasculopathy. It is characterized by narrowing and occlusion of the lumen by intimal proliferation and thrombosis, which leads to ischemia and infarction in the involved organ systems. Its rarity and rapidly fatal course make the disease a difficult diagnostic and therapeutic challenge. PMID- 15779514 TI - Computer briefs: spyware and adware. PMID- 15779513 TI - A patient, an internet article, and thou: challenges of the Internet in clinical practice. AB - This article addresses issues a physician should consider when responding to medical research gathered by a patient from the Internet, discussing both potential medical malpractice liability and offering specific, recommended responses for physicians whose patients conduct online medical research. PMID- 15779515 TI - Cost accounting: making your practice stronger and more efficient. PMID- 15779516 TI - Improving customer service on the phone: a multidimensional effort with a big payback. AB - Telephone communications are the first line of contact for new patients and established patients who require ongoing care. They also represent one of many Achilles' heels for practices. Poor handling of phones can be inefficient and costly. This article outlines The Vancouver Clinic's experience in viewing the problems, identifying the issues, and resolving them with the minimum of expense and personnel disruption. It documents the marked lowering of call abandonment rates and the decrease in call-to-answer times. PMID- 15779517 TI - Coding and billing for postoperative services in the global period by a different physician. PMID- 15779518 TI - Developing a competitive benefits program. AB - Offering your employees the right fringe benefits can help staff morale soar, foster loyalty, and increase the chances that a top-notch job applicant will say yes to your job offer. This article suggests practical ways to offer a competitive benefits program without breaking the bank. It includes guidance about specific benefits and suggests a dozen more extra benefits employees value and a sample cafeteria-style fringe benefits plan. Finally, the article includes guidelines about creating and using your own benefits statement with your staff; along with a model statement form you can use or adapt to your needs. PMID- 15779519 TI - Low-maintenance management: using a simple classification to rate employees' productivity and support requirements. AB - Employees vary in their productivity and their need for guidance, support, and other inputs that may be called "maintenance." This article provides a guide to classifying your team members by a gross ratio of perceived managerial inputs to productivity outputs. It also gives some approaches to understanding your own reactions and dealing with the needs of each group. PMID- 15779520 TI - Should you add a PA or an NP to your practice? AB - Physicians and office managers have long sought solutions to enhancing medical care while controlling costs. One answer is to hire a physician assistant (PA) or nurse practitioner (NP). But which provider is best for the practice? There is no universal staffing solution. It depends on the needs of the patients, the needs of the practice, and the needs of the physician. All three factors should be part of the decision-making equation. This article raises the important questions that should be addressed when deciding whether a PA or an NP is the appropriate provider to add to the staff and offers recommendations on where to go to learn more about both professions to assist a practice in making a decision. PMID- 15779521 TI - Basics of asset protection. AB - Asset protection has become a hot topic for physicians because of the risk of high judgments in medical malpractice cases-judgments that often exceed their policy limits and can force a physician into bankruptcy. In this article we describe some of the background and basics of asset protection. In future articles we will detail some of the protection strategies that can be used. PMID- 15779522 TI - The organized medical staff: should anyone care anymore? AB - Every hospital has an organized medical staff, but of what value is this organizational construct in the current environment? Given the New World Order of quality improvement, patient safety, the malpractice crisis, and the multiple demands physicians struggle to accommodate on a daily basis, it is time to reconsider the role of the medical staff This article looks at the legal basis for the medical staff's role; considers typical medical staff bylaws; and addresses some of today's hottest potatoes between medical staffs and boards, all in light of the new quality era. Based on more than 25 years of working with medical staffs on these issues, and reflecting themes around clinical practice guidelines (CPGs), the author makes the case that although the medical staff is still a vital component of the hospital's mission, both in the bylaws and in its functions, today's staff can be revitalized in ways that can be far more meaningful to physicians and far more likely to propel quality. PMID- 15779523 TI - New considerations in maintaining financial stability. PMID- 15779524 TI - The American Jobs Creation Act: impact on practices. PMID- 15779525 TI - Trends in the consumer price index. PMID- 15779526 TI - From sensory to long-term memory: evidence from auditory memory reactivation studies. AB - Everyday experience tells us that some types of auditory sensory information are retained for long periods of time. For example, we are able to recognize friends by their voice alone or identify the source of familiar noises even years after we last heard the sounds. It is thus somewhat surprising that the results of most studies of auditory sensory memory show that acoustic details, such as the pitch of a tone, fade from memory in ca. 10-15 s. One should, therefore, ask (1) what types of acoustic information can be retained for a longer term, (2) what circumstances allow or help the formation of durable memory records for acoustic details, and (3) how such memory records can be accessed. The present review discusses the results of experiments that used a model of auditory recognition, the auditory memory reactivation paradigm. Results obtained with this paradigm suggest that the brain stores features of individual sounds embedded within representations of acoustic regularities that have been detected for the sound patterns and sequences in which the sounds appeared. Thus, sounds closely linked with their auditory context are more likely to be remembered. The representations of acoustic regularities are automatically activated by matching sounds, enabling object recognition. PMID- 15779527 TI - Putting things into perspective: individual differences in working-memory span and the integration of information. AB - An important function of working memory is the integration of incoming information into an appropriate model of the contextual situation. We hypothesized that individual variability in working-memory function (estimated using Engle's operation-span measure) may lead to differential reactivity to a changing context. We recorded event-related brain potentials (ERPs) and reaction time measures to stimuli embedded in long stimulus series (two auditory discrimination tasks), and examined the participants' responses in relation to how the current stimuli fit with the context generated by the previous stimuli. In both tasks, participants with low working-memory span scores showed larger brain responses as a function of variations in the local stimulus sequence than participants with high span scores. These data suggest that the low working memory span group is more affected by the local stimulus sequence than the high span group, possibly because they are more easily swayed by ongoing changes and are therefore less capable of maintaining their attention on the overall sequence. PMID- 15779528 TI - Local contextual cuing in visual search. AB - Previous research has indicated that covariations between the global layout of search displays and target locations result in contextual cuing: the global context guides attention to probable target locations. The present experiments extend these findings by showing that local redundancies also facilitate visual search. Participants searched for randomly located targets in invariant homogenous displays, i.e., the global context provided information neither about the location nor about the identity of the target. The only redundancy referred to spatial relations between the targets and certain distractors: Two of the distractors were frequently presented next to the targets. In four of five experiments, targets with frequent flankers were detected faster than targets with rare flankers. The data suggest that this local contextual cuing does not depend on awareness of the redundant local topography but needs the redundantly related stimuli to be attended to. PMID- 15779529 TI - Integrating sequential arrays in visual short-term memory. AB - Are sequential visual arrays represented as separate images or as a combined image in visual short-term memory (VSTM)? Proponents of the integration account suggest that an image of the first array is gradually formed and integrated with an image of the second to produce a combined representation. This view is evidenced by successful performance in an empty-cell detection task. In this task, on a 4 x 4 square matrix, 7 locations are occupied on a first array, followed by a variable interval, and then by 8 other occupied locations on a second array. Subjects' success in identifying the remaining empty cell has been taken as evidence for integration. In this study, we show that success in this task can be better accounted for by a convert-and-compare process than by an integration process. We conclude that VSTM only supports limited integration across sequential arrays. PMID- 15779530 TI - The odd-even effect in addition: an analysis per problem type. AB - The odd-even effect is a well documented finding in the literature on mental arithmetic, at least for multiplication. It implies that false answers with the same parity as the correct answer are rejected more slowly than false answers with a different parity. For addition, this effect is not so well documented. The study by Krueger and Hallford (1984) is the only one that investigated odd-even effects for addition. However, they did not investigate odd-even effects per problem, even though there are indications that problem type can modulate odd even effects for multiplication (Lemaire & Reder, 1999). Therefore, we wanted to get more insight into odd-even effects for addition by investigating odd-even effects per problem type. Our results extended the findings of Krueger and Hallford. First of all, we found an interaction between split and problem type. The most important and new result of present study, however, was a strong parity effect for E + E problems. We discuss our results in terms of two alternative explanations for odd-even effects, namely use of a parity rule on the one hand and familiarity with even outcomes on the other. PMID- 15779531 TI - Inconsistent probability estimates of a hypothesis: the role of contrasting support. AB - This paper studies consistency in the judged probability of a target hypothesis in lists of mutually exclusive nonexhaustive hypotheses. Specifically, it controls the role played by the support of displayed competing hypotheses and the relatedness between the target hypothesis and its alternatives. Three experiments are reported. In all experiments, groups of people were presented with a list of mutually exclusive nonexhaustive causes of a person's death. In the first two experiments, they were asked to judge the probability of each cause as that of the person's decease. In the third experiment, people were asked for a frequency estimation task. Target causes were presented in all lists. Several other alternative causes to the target ones differed across the lists. Findings show that the judged probability/frequency of a target cause changes as a function of the support of the displayed competing causes. Specifically, it is higher when its competing displayed causes have low rather than high support. Findings are consistent with the contrastive support hypothesis within the support theory. PMID- 15779532 TI - Escalation of commitment with transparent future outcomes. AB - A frequent case of irrational decision making is the tendency to escalate commitment to a chosen course of action after unsuccessful prior investments of money, effort, or time (sunk costs). In previous research it is argued that escalation does not occur when future outcomes and alternative investments are transparent. Inconsistent with this argument, in an experiment in which undergraduates were presented fictitious investment problems with sunk costs, escalation was demonstrated when full information was given about investment alternatives and estimates of future returns. Thus, it is indicated that people may escalate despite knowing that it will not make them economically better off. A more comprehensive understanding of escalation requires disentangling people's noneconomic reasons for escalation. PMID- 15779533 TI - Clarifying the role of the "other" category in the self-esteem IAT. AB - A. Karpinski (2004) recently criticized Implicit Association Test (IAT) measures of self-esteem, arguing that their measurements of self-associations are compromised by their contrasting self with a putatively extremely negative second category, the nonspecific other. The present data show, to the contrary, that the nonspecific other category in the self-esteem IAT is near neutral in valence. Validity of the self-esteem IAT is most appropriately assessed by examining its correlations with conceptually related measures. That has been done in several previous studies that are reviewed here. The nonspecific other category is only one of several choices for representing the concept of other in self-esteem IATs. Choice of the appropriate other category to contrast with self in self-esteem IATs should be guided by the needs of the research question being addressed. PMID- 15779534 TI - Awareness of pain awareness. PMID- 15779535 TI - Mechanisms associated with unusual orofacial pain. AB - This article presents an overview of possible mechanisms associated with pain perception, with a specific focus on understanding unusual manifestations of orofacial pain associated with nerve insult. It includes recent evidence concerning neurobiological changes that occur in the periphery at tissue and nerve sites, or within the central nervous system, and that may involve chemical and inflammatory responses, sensitization, or alterations of cellular function. Moreover, the contribution of the autonomic nervous system, changes in emotional reactivity and vigilance, the roles of high brain centers such as the basal ganglia (nigro-striatal) system, and the influence of aging and gender, are briefly described. PMID- 15779536 TI - Ultrastructural characteristics of axons in traumatic neuromas of the human lingual nerve. AB - AIMS: To determine the ultrastructural characteristics of axons in traumatic neuromas of the human lingual nerve during the surgical removal of lower third molar teeth and to establish whether any characteristics were different between patients with dysesthesia and patients without dysesthesia. METHODS: Transmission electron microscopy was used to determine the ultrastructural morphological characteristics of human lingual nerve neuromas (n = 34) removed at the time of microsurgical nerve repair. From a sample population of myelinated and nonmyelinated fibers within the neuromas, fiber diameter, myelin thickness, g ratio, and the number of mitochondria per axon were quantified. Comparisons were made with normal control lingual nerve specimens (n = 8) removed at the time of organ donor retrieval. RESULTS: Significant differences in ultrastructural morphology were found between the neuromas and control nerves. The neuromas contained a higher proportion of small (2- to 8-microm diameter) myelinated nerve fibers than controls, and the mean myelinated fiber diameter was significantly lower in neuromas than in controls. Mean myelin sheath thickness was significantly thinner in neuromas (0.6 +/- 0.1 microm) than in controls. However, the g-ratio, which is a measure of the myelination status of the nerve fibers in relation to their diameter, was found to be similar in each group, suggesting a normal process of myelination in the damaged axons. Nonmyelinated axon diameter was also significantly smaller in the neuromas than in the controls, and Schwann cells were found to sheathe more nonmyelinated axons in neuromas than in controls. The ratio of nonmyelinated to myelinated axons was significantly higher in neuromas than in controls. However, no significant differences were found between patients with dysesthesia and those without dysesthesia. CONCLUSION: Damage to the lingual nerve results in marked changes to axon diameter, myelin sheath thickness, and Schwann cell-axon relationships. These ultrastructural changes could contribute to the altered electrophysiological properties of axons trapped within neuromas. However, no significant differences in the ultrastructural characteristics studied were found between specimens from patients with or without symptoms of dysesthesia. PMID- 15779537 TI - Temporal changes in inflammatory mediator concentrations in an adjuvant model of temporomandibular joint inflammation. AB - AIMS: To determine temporal changes in the concentrations found in the temporomandibular joint (TMJ) and trigeminal ganglion of 3 specific classes of inflammatory mediators commonly linked with conditions of joint inflammation. The intent was to determine whether concentrations of the neuropeptide calcitonin gene-related peptide (CGRP), the neurotrophin nerve growth factor (NGF), and the proinflammatory cytokines interleukin-1beta (IL1beta) and tumor necrosis factor alpha (TNF-alpha) are altered in the trigeminal ganglion and TMJ tissues during various stages of adjuvant-induced inflammation of the rat TMJ. METHODS: Adult male rats received bilateral TMJ injection of complete Freund's adjuvant (CFA), while control rats did not receive CFA treatment. The trigeminal ganglion and TMJ tissues were collected at 2 days, and 2, 4, and 6 weeks postinjection and analyzed using either radioimmunoassay or enzyme-linked immunosorbent assay. RESULTS: In the trigeminal ganglion, both CGRP and NGF concentrations were significantly elevated in comparison to controls from 2 days to 4 weeks; however, the patterns of increase differed. Concentrations of each inflammatory mediator were significantly elevated in the TMJ tissues of CFA-injected animals at 2 days and continued to be significantly elevated throughout the 6-week period. CGRP content remained at peak levels from 2 days through 6 weeks, while peak content for NGF, IL-1beta, and TNF-alpha was found at 2 days through 2 weeks. CONCLUSION: The results suggest that the development of CFA-induced inflammation of the TMJ was accompanied by a variable increase in the concentration of different classes of inflammatory mediators in both the trigeminal ganglion and TMJ tissues, which implies that each class of inflammatory mediator may play a significant role during different stages in the onset and exacerbation of the inflammatory process. PMID- 15779539 TI - Generalized joint hypermobility and temporomandibular disorders: inherited connective tissue disease as a model with maximum expression. AB - AIMS: To study the relationship between generalized joint hypermobility (GJH) and temporomandibular disorders (TMD) by assessing prevalence and patient characteristics of TMD in a population of patients with maximum expression of GJH as a symptom of inherited connective tissue disease. In addition, diagnostic reliability of a series of clinical signs indicative of temporomandibular joint (TMJ) hypermobility was tested. METHODS: The study sample consisted of 42 subjects with GJH, 24 with Marfan syndrome and 18 with Ehlers-Danlos syndrome. A subgroup of 27 individuals was selected by age (> or = 18 yrs) and was compared to 40 controls with TMD and normal peripheral joint mobility. TMD diagnoses were assigned to each subject according to the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). RESULTS: In the GJH sample (n = 42), 71.4% of the subjects were symptomatic for TMD. Of those, 13.3% had sought treatment. A myofascial pain diagnosis was made in 69%, disc dislocation with reduction was diagnosed in 85.7%, and TMJ arthralgia in 61.9%. Multiple TMD diagnoses were assigned in 69% of the subjects; of these, 57% had 3 or more subgroup diagnoses. Joint noises (P < .01) and recurrent TMJ dislocations (P < .01) were a frequent finding in adult GJH subjects (n = 27) compared to controls, with symptomatic GJH subjects presenting more and more prolonged dislocation events than asymptomatic subjects (P < .001). TMJ hypermobility signs were expressed significantly more often in GJH compared to controls with TMD and normal joint mobility. CONCLUSION: This study indicates a positive relationship between GJH and TMD. PMID- 15779538 TI - Elevated levels of beta-endorphin in temporomandibular joint synovial lavage fluid of patients with closed lock. AB - AIMS: To investigate the presence of endogenous beta-endorphin, an opioid, in the synovial lavage fluid of the temporomandibular joint (TMJ), and to compare the concentration of 3-endorphin in patients with closed lock with that in symptom free subjects. METHODS: Thirty-eight patients (38 joints) with closed lock diagnosed on the basis of the results of clinical examination and magnetic resonance imaging (MRI) and 11 healthy volunteers (19 joints) were examined. Samples of lavage fluid were obtained prior to arthrocentesis by washing the joint with saline. Samples were assayed for beta-endorphin by an enzyme immunoassay, and concentrations of protein were measured by a bicinchoninic acid assay. Subjective pain was assessed by patients using a visual analog scale. Bone changes in the condyle were assessed by MRI, and synovitis was assessed on the basis of arthroscopic findings. RESULTS: beta-endorphin was present in the synovial fluid of the TMJ, and the concentration was significantly higher in patients with closed lock of the TMJ compared to symptom-free volunteers. The beta-endorphin levels were not, however, significantly correlated with clinical parameters in the patients. CONCLUSION: The study results support recent findings that some opioids and their receptors exist not only within the central nervous system but also in the TMJ region, and that opioid concentrations are higher in patients with pain and dysfunction of the TMJ. PMID- 15779540 TI - Is there a difference in the reliable measurement of temporomandibular disorder signs between experienced and inexperienced examiners? AB - AIMS: To determine whether there is a difference in terms of reliability between experienced examiners and inexperienced examiners in the measurement of signs of temporomandibular disorders (TMD). METHODS: A total of 27 patients seen for treatment of TMD were rated blindly and in random sequence by 2 experienced and 2 inexperienced examiners. The examiners participated in a 4-hour calibration session on the day preceding the reliability study. Both experienced and inexperienced examiners participated in the calibration session to reduce the effect of examiner subjectivity and allow the study focus to be on the effect of experience. The rating followed the Research Diagnostic Criteria for Temporomandibular Disorders and included mandibular movements, joint sounds, and digital palpation of muscles and joints. Intraclass correlation coefficients and kappa statistics were calculated to estimate interrater reliability. The Wilcoxon signed rank test was performed to test for differences between experienced and inexperienced examiners' results, and the Friedman test was used for differences between all 6 examiner combinations. RESULTS: Excellent overall reliability was found for vertical mandibular motions, acceptable reliability was found for the summed muscle palpation pain sites, and moderate to poor reliability was found for excursive movements, joint sounds, and single muscle palpation pain sites. No significant differences in the measurement results could be found between the experienced examiners and the inexperienced examiners. CONCLUSION: Examiner calibration rather than professional experience seems to be the most important factor for reliable measurement of TMD symptoms. PMID- 15779541 TI - Diagnoses based on the Research Diagnostic Criteria for Temporomandibular Disorders in a biracial population of young women. AB - AIMS: To compare the clinical characteristics of diagnostic subtypes of temporomandibular disorders (TMD) based on the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) in terms of physical findings (Axis I) and psychosocial findings (Axis II) among Caucasian and African American young women. An ancillary goal was to assess the value of using self-reported TMD pain as a screening tool compared to RDC/TMD examinations. METHODS: A biracial community sample (n = 830) of young women 19 to 23 years old was screened for facial pain with the Chronic Pain Grade questionnaire. Patients were considered to be putative cases of TMD if they reported facial pain present within the last 6 months; putative controls had no facial pain history or jaw symptoms. Women with facial pain more than 6 months ago and jaw symptoms (jaw symptom-past pain, JSPP group) were added. 129 women were clinically examined for TMD diagnosis for final confirmation of case-control status. RESULTS: 41 of 43 Caucasian and 11 of 18 African American putative cases were confirmed as cases; 9 of 27 Caucasians, but 0 of 17 African Americans from the JSPP group were confirmed as cases. All 24 putative controls were confirmed as controls. Based on RDC/TMD Axis I, 80% of 61 cases were muscle-related diagnoses, 33% as disc-related diagnoses, and 48% as arthralgia/arthritis/arthrosis. Based on Axis I, there were no significant differences in diagnoses between African American and Caucasian women. Based on Axis II, cases had significantly greater depression (P = .002) and somatization with pain (P < .001) than controls as expected. African Americans had significantly greater somatization with pain than Caucasians (P = .020). There were no other significant racial differences. CONCLUSION: Among young women reporting facial pain, clinical TMD subtypes, pain impact, treatment utilization, and additional characteristics other than somatization with pain were similar between races. A high percentage of these young non-clinical cases presented severe depression and somatization. PMID- 15779542 TI - Jaw clenching modulates sensory perception in high- but not in low-hypnotizable subjects. AB - AIMS: To investigate the effect of jaw clenching on the sensations evoked at segmental and nonsegmental levels by painful and nonpainful stimuli and in relation to hypnotic susceptibility. METHODS: The effect of jaw clenching on painful and nonpainful sensations on the face and leg was studied in high hypnotizable (HH) and low-hypnotizable (LH) subjects. Sixteen healthy subjects were selected and assigned to either the HH group (n = 8) or the LH group (n = 8). Painful and nonpainful electrical stimuli were delivered in random order to the face and leg. The subjects rated the intensity of the evoked sensation on a visual analog scale (VAS) while clenching or not clenching their jaw. RESULTS: Jaw clenching significantly attenuated the VAS sensory ratings of all the subjects under various conditions (F(1-31) = 6.15, P < .02). When the HH and LH subjects were analyzed separately, jaw clenching was found to be effective in reducing sensations only in the HH subjects (F(1-15) = 8.30, P = .01), only those evoked in the face (segmental level), and only those evoked by nonpainful stimuli (tied Z = 2.52, tied P < .02). CONCLUSION: Sensory modulation produced by jaw clenching may be related to hypnotic susceptibility. On the whole, jaw clenching had a weak, local effect in modulating sensation, in contrast to its known widespread effect on motor behavior. PMID- 15779543 TI - Dominant form of arthrogryposis multiplex congenita with limited mouth opening: a clinical and imaging study. AB - AIMS: Arthrogryposis multiplex congenita (AMC) is characterized by congenital contractures and joint deformities, but there are only a few reports of temporomandibular joint (TMJ) involvement. The objective of this investigation was to study the cause of limited mouth movement in this disease. MATERIALS AND METHODS: Four individuals from a family affected by AMC over 5 generations were examined clinically and by magnetic resonance imaging (MRI) and 3-dimensional computerized tomography (3D-CT). RESULTS: The CT scans of the 4 individuals showed hyperplasia of the coronoid process protruding into the infratemporal fossa in 2 of them and cranially to the zygomatic arch in the other 2; the hyperplasia was associated with mechanical limitation of the mouth opening. The MRI showed a disc displacement with reduction in 1 patient and a disc displacement without reduction in another; disc displacement could not be evaluated because of the limited mouth opening in the other 2. The condyle-disc complex of these last 2 individuals could only rotate. The MRI on T2-weighted images showed disc hyposignal in all cases but no alterations in the masticatory muscle tissue. The pedigree of the family suggests an autosomal dominant form of inheritance. CONCLUSIONS: The restriction of mouth opening in the 4 individuals affected by AMC was likely due to osseous dysplasia. PMID- 15779544 TI - Myopain 2004: Sixth World Congress on myofascial pain and fibromyalgia. July 18 22, 2004, Munich, Germany. PMID- 15779545 TI - Slights. PMID- 15779546 TI - Come ye to the waters. PMID- 15779547 TI - Primary prevention for older persons. PMID- 15779548 TI - Cognitive screening of older adults. PMID- 15779549 TI - Another perspective on the memantine graphs: clinical vs. statistical significance. PMID- 15779550 TI - Safe and effective drug therapy in older adults. AB - The last two decades have witnessed the introduction of major advances in drug therapy for many chronic diseases. Despite an increasing number of therapeutic choices, basic challenges remain when using new drugs in the elderly. It isessential: (1) to consider whether a new drug is truly safer, more effective or worth the extra expense compared to established medications, (2) to monitor patients for drug related toxicities, and (3) to avoid polypharmacy and the prescribing cascade. PMID- 15779551 TI - Deep venous thrombosis as a consequence of benign prostatic hypertrophy. PMID- 15779552 TI - Prevention of falls in older adults. PMID- 15779553 TI - Influenza 2004-2005. PMID- 15779554 TI - Miriam Hospital and Rhode Island Hospital morbidity and mortality conferences, two cases of sudden weakness. PMID- 15779555 TI - Hospital community benefits in 2003. PMID- 15779556 TI - Colorectal cancer screening resources in Rhode Island. PMID- 15779557 TI - First-ever HIPAA conviction highlights differing views of HIPAA's civil and criminal penalties. PMID- 15779558 TI - Expert witnesses. PMID- 15779559 TI - Contingency fee legal actions in the U.S.A. PMID- 15779560 TI - The passageways of medicine. PMID- 15779561 TI - Community spirit. PMID- 15779562 TI - Maggie Stubbs. PMID- 15779563 TI - Changing times. PMID- 15779564 TI - Going global. PMID- 15779565 TI - Managing the aftershock. PMID- 15779566 TI - Ending the paper chase. PMID- 15779567 TI - Managing change: cultural diversity in the NHS workforce. PMID- 15779568 TI - Mentoring and the value of observation. PMID- 15779569 TI - Gene delivery into the chicken embryo by using replication-competent retroviral vectors. AB - Rous sarcoma virus (RSV)-derived retroviral vectors have allowed for efficient gene transfer into the chicken embryo which is a classical model for studying vertebrate development. Current evidence reveals that this method can be used for regionally restricted expression, inducible expression, and for interfering with endogenous gene function, suggesting that gain-of-function and loss-of-function strategies for specific genes can be achieved spatially and temporally in the avian embryo. Thus, retroviral-mediated gene transfer into the chicken embryo coupled with a wide variety of strategies is now an important tool to address specific biological questions in the vertebrate. PMID- 15779570 TI - Pathoanatomic investigation of cervical spondylotic myelopathy. AB - Multiple dural sac and spinal cord indentations are often observed on MRI in cervical spondylotic myelopathy. However, it is rare that all of the indented levels contribute to clinical symptoms. Pathological changes in cervical compression myelopathy have previously been reported. Still the critical degree of spinal cord compression needed to induce pathologic changes is unknown. To clearfy this matter the relationships between the spinal cord as well as the dural sac indentations, and pathological changes of the cervical spinal cord were investigated in cadavers. Sixty-eight cadavers were used for this study. The dural sac and the spinal cord were observed especially regarding presence of indentations. The spinal cord was removed from the specimens in order to perform histopathological examination. The indentations of the dural sac and the spinal cord were observed at C4/ 5, C5/6, C6/7 and C3/4 intervertebral levels in order of incidence. However, all of the dural sac indentations were not correlated with spinal cord indentations. Pathological changes in the spinal cord were observed in two specimens with less than 30% of the AP compression ratio. One specimen with 20.9% in the AP compression ratio had remarkable histopathological changes. In the other specimen with 29.6% in the AP compression ratio, diffuse demyelination was distributed in the lateral white matter. The results indicate that the critical degree of the AP compression ratio is 30% to induce histopathological changes in the spinal cord. If a spinal cord indentation in the patient with cervical spondylotic myelopathy on imaging, i.e. MRI, show less than 30% in the AP compression ratio, the clinical symptoms, i.e. numbness, tickling and paresthetic pain, may not be improved after the surgery because of some histopathological changes in the spinal cord. PMID- 15779572 TI - Histological distribution and ultrastructural features of immunoreactive terminals against RT97, a monoclonal antibody to a 200 kD neurofilament, in the spinal dorsal horn of a rat. AB - Localization and ultrastructural features of immunoreactive fibers and terminals against RT-97, a mouse monoclonal antibody that recognizes subunit of a 200-kD neurofilament, were examined in the spinal dorsal horn of adult rats. Under a light-microscope, many RT-97 immunoreactive fibers were detected in the dorsal root, collaterals of the dorsal root in the dorsal funiculus, and laminae III and IV in the dorsal horn. Few immunoreactive fibers were found in laminae I and II. Electron microscopic observation demonstrated that almost all RT-97 immunoreactive fibers in the dorsal root were myelinated, and unmyelinated fibers immunonegative. The immunoreactive fibers entered into the dorsal horn passing through the collaterals of the dorsal root along the superficial gray lamina. In the dorsal horn, these fibers ascended into and then terminated in lamina II. RT 97 immunoreactive central terminals were semicircular or ellipsoid in appearance and contained many flat-type presynaptic vesicles. Some terminals made synaptic contact with dendritic profiles in lamina II. Our present results indicate that RT-97 is a useful marker for ultrastructural examination of terminals served by non-nociceptive A-fibers. PMID- 15779571 TI - A comparative study on contractile responses of rabbit and guinea pig vasa deferentia to electrical field stimulation. AB - Contractile responses of rabbit and guinea pig vasa deferentia to electrical field stimulation (EFS) are compared. A muscarinic receptor blocking agent, 1 microM atropine markedly reduced phasic and tonic contraction induced by EFS (20 Hz, 0.5 msec, 30 V, for 30 sec) in rabbit vas deferens, while it only slightly depressed those in guinea pig vas deferens. Further addition of an adrenergic alpha1 receptor blocking agent, 1 microM prazosin markedly depressed the second tonic contraction in both rabbit and guinea pig vasa deferentia. In the presence of atropine and prazosin, further addition of a P2X purinoceptor desensitizing agent, 10 microM alpha,beta-methylene ATP (alpha, beta-MeATP) abolished the residual phasic contractile response in guinea pig vas deferens, while it partially depressed that in rabbit vas deferens. The administration of 10 microM alpha,beta-MeATP in the absence of atropine and prazosin markedly potentiated the phasic contractile response of rabbit vas deferens to EFS, while it depressed that of guinea pig vas deferens. Contractile response of rabbit vas deferens to alpha,beta-MeATP was more potent than those of ATP and 2-methyl-thioATP (2-Me thioATP), while these nucleotides had almost same potency in guinea pig vas deferens. These findings may indicate that contribution of cholinergic, adrenergic and purinergic neurotransmission to the contractile response of rabbit vas deferens to EFS is different from that of guinea pig vas deferens. PMID- 15779574 TI - Molecular medicine: the way forward. PMID- 15779573 TI - A case of annular pancreas in a male adult. AB - Annular pancreas is a rare congenital anomaly, which consists of a ring of pancreatic tissue partially or completely encircling the descending portion of the duodenum. We reported a case of symptomatic annular pancreas in a 40 year old man admitted to our hospital complaining of abdominal pain, nausea and vomiting without body weight loss in January 2000. The patient underwent laparoscopic cholecystectomy for acalculous cholecystitis in September 1996. Initially, he was diagnosed with duodenal stenosis due to a duodenal ulcer scar, but laboratory data showed no abnormalities. His symptoms did not improve with medication or endoscopic balloon dilatation. Duodenograpy revealed a narrow segment with a smooth mucosal surface in the 2nd portion of the duodenal loop in the duodenum, and a computed tomography (CT) scan demonstrated a thickened pancreas head around this narrow segment. We were therefore able to diagnose annular pancreas. A duodeno-duodenostomy was performed in March 2000. The patient's postoperative course was uneventful, and he was discharged from our hospital on the 19th postoperative day. Although define diagnosis of annular pancreas is frequently made at laparotomy, the development of a recurrent imaging modality might assist in the preoperative diagnosis. PMID- 15779575 TI - Impact of sexual health course on Malaysian university students. AB - A sexual health course was offered and taught by academic staff from the Faculty of Medicine, University of Malaya during semester II of every year as a university elective course to other university students apart from medical students. The course covered a wide range of topics: adolescent sexuality, family planning and pregnancy, violence against women, alternative sexual behavior, physiology of sex, sex and the disabled, gender bias in sexuality, relationship and marriage, sexual dysfunctions, clarification of sexual attitudes and STDs and AIDS. The Sexual Knowledge and Attitude Test (SKAT-II) was used to measure students' pre- and post-course scores on sexual knowledge and attitudes. Fifty four students who completed both the pre- and post-course tests showed a significant change in sexual knowledge and their attitudes towards sexual myths and autoeroticism. Sexual knowledge was also positively correlated with age, heterosexual relations, autoeroticism and sexual myths scores. However, sexual knowledge is negatively related to religiosity and the influence of religious beliefs on one's attitudes towards sexual matters. This study showed that the sexual health course offered does have a positive impact in increasing one's knowledge and changing one's attitudes towards sexual issues. PMID- 15779576 TI - Nerve injuries in anterior shoulder dislocations. AB - A retrospective study was conducted to determine the incidence of nerve injuries in anterior dislocations of the shoulder and to determine if a neurological examination is routinely performed in such cases. One hundred and fifteen shoulder dislocations in 100 patients (74 males, 26 females; mean age 35+/-18 years) were reviewed. Eighty-seven patients were examined for neurological injuries prior to reduction and 8 of these patients (9.2%) were found to have sustained neurological injuries. Following reduction, neurological examination was performed in 85 patients. Three patients who were not examined initially prior to reduction were subsequently found to have nerve injuries after reduction. Seven patients (7%) were not examined for neurological injury both before and after reduction of the dislocation. Eleven patients were found to have sustained nerve injuries in the final analysis. Axillary nerve injuries were the most common, occurring in 8 patients. In conclusion, nerve injuries are common in shoulder dislocations. The importance of performing and documenting the findings of neurological examination anterior shoulder dislocations needs to be reiterated. PMID- 15779577 TI - An epidemiological investigation of an outbreak of leptospirosis associated with swimming, Beaufort, Sabah. AB - From October 13 to October 21, 1999, a total of 46 males, aged 8 to 19 years, were admitted to Beaufort Hospital after swimming in the creek near an oil palm plantation in Kampung (Kg) Kebatu, Beaufort. Thirty (30) presented with symptoms including fever, vomiting, bodyache, giddiness, headache, chest pain and cough, while 16 others, were asymptomatic. One, a 15 year old boy, died from haemorrhagic shock secondary to pulmonary haemorrhage. The onset of the illness was from 11 October to 19 October 1999. A case-control study found that the outbreak was associated with swimming in the creek (p<0.0001). A total of 44 paired sera samples were sent for microscopic agglutination test (MAT), 5 pairs showed sero-conversion, 3 pairs had 4 fold rise in titre and 18 pairs were positive at >320. The findings indicated that Leptospirosis was the cause of the outbreak of this illness and the contaminated creek water was the source of the infection. The occurrence of flooding and stagnation in the creek following the heavy rainfall during the first week of October 1999 could have contributed to the timing of the outbreak. PMID- 15779578 TI - Blood pressure and associated cardiovascular risk factors in Iran: Isfahan Healthy Heart Programme. AB - This study was done to estimate the prevalence of high blood pressure (BP) in treated and non-treated subjects with respect to age and gender and its association with other cardiovascular risk factors in Iran. This cross sectional study was performed in three cities of Iran on participants over 19 years at 2002. First a questionnaire consisting of demographic details, drug intake and smoking status was filled. Then physical examination including systolic and diastolic blood pressure (SBP, DBP), body mass index (BMI) and waist to hip circumference (WHC) was performed. Fasting blood sample was drawn for sugar (FBS), total cholesterol (TC) and triglyceride (TG) and a 2-hour postprandial glucose was also measured. In this study performed on 12494 subjects, 48% were males and 52% females. The mean age of men and women was 38.99+/-15.30 and 38.80+/-14.54 years respectively. The prevalence of high BP in men and women was 15.6% and 18.8% respectively. The prevalence of high BP was higher in women than in men, except in the younger age classes. Overall 26.7% of hypertensive men and 47.7% of hypertensive women were on anti-hypertensive pharmacological treatment. Among the treated patients, BP was under control in 6.4% of the men and 13.8% of the women. In 86.5% of men with high BP and 89.3% of women with high BP, at least one other cardiovascular risk factor was present and its prevalence increased with age in both genders. BMI >25 (especially abdominal obesity) was the most frequent associated risk factor (41.9% in male, 59% in female). Except for smoking, the prevalence of each cardiovascular risk factor increased with the severity of hypertension, except in young women. The prevalence of high BP- even in treated subjects- is high in Iran. Many subjects with high BP have at least one other associated cardiovascular risk factor. These data emphasize the necessity of implementing community-based interventions. PMID- 15779579 TI - Presence of allelic loss and PTEN mutations in malignant gliomas from Malay patients. AB - Loss of heterozygosity (LOH) on several loci and mutations on PTEN tumor suppressor gene (10q23.3) occur frequently in sporadic gliomas. We have performed polymerase chain reaction (PCR)-LOH analysis using microsatellite markers and single-stranded conformational polymorphism (SSCP) analysis to determine the incidence of allelic losses on chromosome 10q, 9p, 17p and 13q and mutations of exons 5, 6 and 8 of the PTEN gene in malignant gliomas. Twelve of 23 (52.2%) malignant glioma cases showed allelic losses whereas 7 of 23, (30.4%) samples showed aberrant band patterns and mutations of the PTEN gene. Four of these cases showed LOH on 10q23 and mutations of the PTEN gene. The data on LOH indicated the involvement of different genes in gliomagenesis whereas mutations of the PTEN gene indicated the role of PTEN tumor suppressor gene in the progression of glioma in Malay population. PMID- 15779580 TI - Presence of telomerase activity with undetectable p16 gene mutation in Malaysian patients with brain tumor. AB - Recent study has shown that activation of the telomerase and p16 gene mutation are both necessary for tumorigenesis. Our objectives were to detect telomerase activity and investigate the possibility of p16 gene mutations in various types of brain tumor. We analyzed 23 tumor tissues collected in 2000 to 2002. Telomerase activity was detected by a TRAP assay using a TRAPEZE Telomerase Detection Kit (Intergen, Co). PCR-SSCP (Single Strand Conformation Polymorphism) analysis was performed to screen for p16 gene mutation at exon 1 and 2. The activity was detected in 26.1% of the brain tumor samples and mostly present in high-grade tumors. There was a significant association between telomerase activity status and tumor grade but not with patient criteria. Telomerase activity was detected in the analyzed tumors, supporting the fact that activation of telomerase is an important feature for tumorigenesis. There was no mobility shift of p16 gene using SSCP and suggested no mutation at exon 1 and 2 occurred in all samples. These results suggest that another mechanism of p16 gene alterations could be involved and associated with detectable telomerase activity in the progression of tumors. PMID- 15779581 TI - Ultrasound in the assessment of the palpable breast mass. AB - The aim of the study was: to obtain the profile of patients (with regards to age and family history of breast cancer) with a palpable breast mass. To determine the validity of ultrasound in the assessment of the palpable breast mass by determining the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of ultrasound in distinguishing a malignant mass. To determine the most discriminating ultrasound characteristics for differentiating benign and malignant masses. Seventy patients who had fine needle aspiration cytology of a palpable breast mass were subjected to an ultrasound assessment of the mass. The ultrasound findings were classified as benign, indeterminate or malignant. These findings were then compared with either the cytology or histology results in cases that eventually had surgical excision. The age of the patients ranged from 15 to 66 years old The majority was in the third and fourth decades with an average age of 25 years. The 8 patients with a proven malignant breast mass were aged between 39 and 66 years old. They did not have any family history of breast cancer. Only 4 patients had a family history of breast carcinoma and all proved to have a benign breast lesion. Ultrasound had a sensitivity of 100%, specificity of 85.7%, positive predictive value of 50%, negative predictive value of 100% and accuracy of 87.5% for distinguishing a malignant mass. For benign masses: 93.7% had well-defined margins, 81.3% had homogenous internal echoes, 91.7% had depth-width ratio of less than 1.0 and 89% were compressible. For malignant masses: 87.5% had either ill-defined or irregular margins, 87.5% had inhomogenous internal echoes and mixed posterior echoes, and 100% were incompressible. The majority of patients with a palpable breast mass were aged below 40 years old. Most of the patients with a malignant breast mass were aged 40 years and older. Neither a positive nor a negative family history of breast cancer had any significance on outcome. Ultrasound had high sensitivity, specificity and accuracy in distinguishing a malignant mass. The most discriminating benign ultrasound characteristic was compressibility. The most discriminating malignant ultrasound characteristic was ill-defined and irregular margins. PMID- 15779582 TI - The utilization of an index for serum globulin compensation in diseases associated with decreased serum albumin. AB - The albumin globulin ratio (A/G ratio) is meant to represent the ratio of alterations in serum proteins, since, in liver disease, globulins (G) rise following serum albumin (SA) decrease. pathophysiological value, its' use has been limited. Alternatively, we have developed an index, the globulin compensation index (GCI) to measure the changes in serum globulins when albumin is decreased. The index is calculated as follows: G - 25 / 35 - SA. The GCI has been tested using retrospective patients' data from the Hospital Universiti Sains Malaysia. Analysis of the data shows that the GCI may be of potential value in showing the actual serum protein status, especially in cases where globulins are decreased along with albumin. Furthermore, globulin rise in cases with reduced albumin was found in 72.3% of cases of hepatic diseases, whereas this finding occurred in up to 32.3% of cases of non-hepatic, systemic diseases. PMID- 15779583 TI - Knowledge and attitudes towards tuberculosis among the people living in Kudat District, Sabah. AB - A multistage random sampling method was used to select the community sample in the district of Kudat, Sabah. A total of 205 respondents from 210 selected houses were interviewed using a standardised questionnaire to ascertain their knowledge and attitude towards tuberculosis. Generally the knowledge about tuberculosis was poor. The well known symptoms that the respondents knew were coughing blood (46.2%), cough (37.1%), loss of weight (34.5%), and loss of appetite (32.0%). Only 51% thought that the disease was caused by germs and it was transmitted by air. TB sufferers were thought to be dirty (22%) and the majority (51%) were not keen to mix with TB patients. Although more than 90% of the respondents considered TB as socially acceptable within their family and community, a large proportion (41%) expressed that getting TB was embarrassing, 4% said it was a disgrace to the family, and 16% said that it was too sensitive to discuss about it. These behaviours suggest that at the private level, the respondents were still perceived negative social attitudes towards tuberculosis. PMID- 15779584 TI - Analysis of the survival motor neuron and neuronal apoptosis inhibitory protein genes in Malay patients with Spinal Muscular Atrophy. AB - In Malaysia, Spinal Muscular Atrophy (SMA) is diagnosed based on clinical observation with or without muscle biopsy. Molecular analyses of the SMA-related genes have not been available so far. In this preliminary study, we searched for homozygous deletion of Survival Motor Neuron (SMN1) and Neuronal Apoptosis Inhibitory Protein (NAIP) genes in Malay patients with SMA and found homozygous deletion of SMN1 exon 7 and 8 in all the patients while homozygous deletion of NAIP exon 5 was detected in only our type 1 patients but not in the type 3 patient. To the best of our knowledge, these are the first SMA cases diagnosed at the molecular level in Malaysia. PMID- 15779586 TI - Atopic sensitization of children with rhinitis in Malaysia. AB - Atopy is defined as the genetic propensity to develop immunoglobulin E antibodies in response to exposure to allergens and assessed by skin prick test (SPT) responses to common allergens, which may contribute to the development of the clinical disorders (phenotype). Although it is generally agreed that atopy is an important risk factor for allergic diseases such as asthma, rhinitis, and eczema, the extent to which atopy accounts for these diseases is controversial. One hundred forty one children (up to 12 years) were skin prick tested to evaluate 16 foods common to the Malaysian diet and 4 common aeroallergens. Eighty-five percent of patients had positive SPT reactivity. The most commonly implicated aeroallergen and food allergen was house dust mite (HDM) and Prawn. Seventy percent had positive SPT reactivity results to HDM and 24.8% to prawns. Fifty five percent were positive to more than one allergen and 17.7% positive to single aeroallergen. The prevalence of atopy in children with history of eczema was 90%. The incidence of HDM and food allergy especially crabs and prawns, is significantly greater in Malaysian Children with rhinitis symptoms. PMID- 15779585 TI - P53 expression in colorectal carcinoma: the University of Malaya Medical Centre's experience. AB - Loss of P53 function is regarded as one of the critical steps in colorectal carcinogenesis. This study determines the P53 expression pattern of colorectal carcinoma in a cohort of 116 local patients. There was no significant relationship between overexpression of P53 with tumour stage (p=-0.209, chi square test) and grade (p=0.877, chi square test). Survival analysis using Kaplan Meier procedure did not show significant relationship between P53 positivity with overall recurrence-free and survival outcome (p=0.3322 and 0.921 respectively; log rank test). Long-term follow-up may give a better evaluation on the prognostic value of P53 overexpression in colorectal carcinoma. PMID- 15779587 TI - Large fusiform posterior cerebral artery aneurysm (P2 segment): successful trapping and excision via subtemporal approach. AB - A 62-year-old man presented with a 2-month history of right occipito-parietal headache and worsening ipsilateral ptosis. CT scan and cerebral angiography revealed a large fusiform aneurysm of the P2 segment of the right posterior cerebral artery (PCA). The aneurysm was successfully trapped and subsequently excised via sub temporal approach leaving no additional neurological deficits to our patient. PMID- 15779588 TI - Lingual thyroid--a lesson to learn. AB - We present a case of a middle-aged woman with a mass in the posterior third of the tongue which was diagnosed as a tongue haemangioma. The tumour was successfully excised via a midline mandibular osteotomy and tongue splitting approach. The histopathology examination, however, revealed the 5x4 cms mass to be a lingual thyroid. The salient features of this unusual presentation of a thyroid enlargement will be discussed. PMID- 15779589 TI - A very rare and rapidly fatal case of Chromobacterium violaceum septicemia. AB - Chromobacterium violaceum infection is rare but causes a high mortality rate particularly in immunosuppressed persons. Since its clinical presentation is non specific and the diagnosis basically relies upon blood culture and sensitivity, this infection should be considered among the organisms targeted empirically for antibiotic therapy when a cellulitis or rapidly progressive illness follows exposure to water or soil. This is a case of fulminant septicemia caused by this rarely encountered organism. PMID- 15779590 TI - Skin-sparing mastectomy. AB - Skin-sparing mastectomy is still in its infancy in Malaysia. The option of skin sparing mastectomy is rarely given to patients as many general surgeons perform the conventional mastectomy. This could also be compounded by the lack of awareness amongst the local surgeons on the safety, surgical technique and treatment outcome of this relatively new procedure. This case report demonstrates the feasibility of this procedure performed on a Malaysian patient with a comparable outcome of those reported in the Western countries. PMID- 15779591 TI - Heterotopic pancreas in the stomach. AB - A patient diagnosed with gastrointestinal stromal tumour of the stomach underwent gastrectomy. Post-operatively, histopathology was reported as heterotopic pancreas of the stomach. Heterotopic pancreas is very rare and though most of them appear asymptomatic, surgical resection is advisable. PMID- 15779592 TI - Dysphagia as a side effect of botulinum toxin injection. AB - Dysphagia is a known adverse effect of botulinum toxin injection into the cervical region for dystonia. We present an unusual case of dysphagia arising from injection into the orbicularis oculi muscle, which has hitherto not been described. We postulate that her dysphagia was caused by distant side effects of botulinum toxin due to repeated injections. We recommend that clinicians should restrict the frequency of injections to as few life-time doses of the toxin as possible for adequate management of spasm. The practice of re-injecting patients routinely every three months, or at the first return of mild spasms should be discouraged. PMID- 15779593 TI - Behcet's disease with vascular complications. AB - A young gentleman presented with a history of multiple pseudoaneurysms of the right carotid artery, left radial artery, right femoral artery as well as deep vein thrombosis involving the right femoral vein. A diagnosis of Behcet's disease was made on the basis of his history of recurrent oral and genital ulceration with characteristic eye lesion. PMID- 15779594 TI - Dentigerous cyst of the maxillary sinus in a child. AB - Dentigerous cyst in maxillary sinus, especially in children, is uncommon in Malaysia. Few cases of dentigerous cyst in maxillary sinus in children have been reported in the medical literature. According to Tay AB et al, dentigerous cyst was accounted for 2.3% of the 20 most common diagnosed oral tumors in Singapore form year 1993--1997. This report illustrates a case of the dentigerous cyst in the maxillary sinus, resulting in significant facial swelling, which was managed by endoscopic marsupialization of the cyst. PMID- 15779595 TI - Congenital nasal encephalocele--a review of surgical techniques. AB - We report a case of a 6 month old baby boy who had congenital nasal encephalocele, repaired via the traditional staging procedure. The surgical techniques and procedures are described and discussed. PMID- 15779596 TI - Solid pseudopapillary tumor of the pancreas in a young Malay woman. AB - Solid pseudopapillary tumors of the pancreas are very rare, low-grade malignant potentially curable neoplasms. Despite having non-specific symptomatology, they have typical features such as being more common in young women, and classically presenting as large abdominal masses. Accurate diagnosis is important because long-term survival hinges on complete resection of the tumor. PMID- 15779597 TI - Neisseria meningitidis isolates with moderate susceptibility to penicillin. AB - Neisseria meningitidis is one of the important cause of meningitis and has been extremely susceptible to penicillin. Nevertheless, moderately penicillin resistant strains have been reported in some parts of the world. To our knowledge, there is no such report in Malaysia. We report two clinical isolates that were found to have MICs of decreased susceptibility to penicillin by the agar dilution method. PMID- 15779598 TI - Differences in the pattern of gastric carcinoma between north-eastern and north western peninsular Malaysia: a reflection of Helicobacter pylori prevalence. AB - A retrospective study on demographics of gastric carcinoma was conducted in Hospital Pulau Pinang (HPP) with the aim of comparing it to a previous study done in Hospital Universiti Sains Malaysia (HUSM), Kelantan. The incidence of gastric carcinoma was much higher in Penang compared to Kelantan. It was commonest in males and Chinese. The incidence and site of gastric carcinoma closely parallels Helicobacter pylori infection rates. This was evidenced by the higher incidence and non-cardia location of gastric carcinomas in an area with higher H. pylori infection rates (HPP) compared to a much lower incidence and preponderance of cardia tumours in HUSM where the H. pylori infection rate is exceptionally low. PMID- 15779599 TI - Age related visual impairment in the elderly. AB - Visual impairment among the elderly is a major health problem. With advancing age, the normal function of eye tissues decreases and there is an increased incidence of ocular pathology. Demographic studies have shown that age is the best predictor of blindness and visual impairment. The most common causes of age related visual impairment in the elderly are presbyopia, cataracts, age related macular degeneration, primary open angle glaucoma and diabetic retinopathy. Untreated visual impairment leads to physical handicap, increased incidence of fall, depression, social isolation and dependency. Active screening for visual loss in the elderly should be part of the health examination. The elderly should be encouraged to come for formal 1-2 yearly eye assessment for early detection of visual impairment and to treat all associated problems in order to prevent permanent visual loss. PMID- 15779600 TI - Fulminant babesiosis manifested soon after coronary bypass surgery. PMID- 15779601 TI - Adenocarcinoma of the prostate metastatic to the testis via lymphatic invasion: a case report. PMID- 15779602 TI - Successful cesarean section performed with inadvertent subdural anesthesia. PMID- 15779603 TI - Adequacy of iodine nutrition in the United States. AB - Concerns have been raised about the adequacy of iodine nutrition in the United States despite recent NHANES III data indicating that iodine intake remains generally adequate. Such concerns probably reflect misunderstanding of definitions of iodine deficiency. We review current criteria for iodine deficiency, the reasons for variability of urine iodine determinations, and their relation to interpretations of NHANES data. Although NHANES data indicate that iodine nutrition in the United States is adequate, the possibility remains that those who adhere to restricted diets may have reduced intake of iodine. Because of such possibilities, patients' diets and use of dietary supplements should be explicitly considered as a part of routine medical care. PMID- 15779604 TI - Fascial closure: is there a standard practice? AB - A review of the literature was performed with respect to suture type and best practice in securing fascial closure. A Medline search of the literature under "fascial closure," "fascia," and "wound healing" was performed. Studies comparing the types of sutures and the different techniques were reviewed. A critique of meta-analysis studies was performed. Those articles not prospective in nature or including extraneous topics were eliminated. Research studies were selected on a nonbiased basis, comparing suture type and techniques for closure of fascia. Results were based on review of the literature. No valid conclusion could be drawn pertaining to mode of closure. Numerous suture types as well as different modalities of closure were found reliable. PMID- 15779605 TI - The use of prophylactic amiodarone after cardiothoracic surgery reduces the incidence of atrial fibrillation. PMID- 15779606 TI - Stem cell research: ethical concerns--'great expectations'. PMID- 15779607 TI - What I am hearing from physicians. PMID- 15779608 TI - Pride, shame, and the oath. PMID- 15779609 TI - [Bronchial asthma (in the adult)]. PMID- 15779610 TI - [Dysphagia: clinical management in diagnostics and therapy]. AB - The oral nutrition is a complex procedure of different mechanisms. It saves the survival and is part of the social integration. That's why dysphagia seems to be a serious symptom. The aetiology is multiple and is often not recognized. With a profound diagnostic algorithm a lot of aetiologies can be discovered. Treatments can be conservative or surgical. The aim of therapy is to develop techniques of swallowing to reach an oral nutrition and to reintegrate patients in their social net. It is important to recognize early signs. The general practicer is an important initiator to introduce a differential diagnostic procedure. PMID- 15779611 TI - [Neuro-endocrine tumors of the gastrointestinal tract: epidemiology, classification, prognosis, diagnosis and therapeutic modalities]. AB - The neuro-endocrine tumors of the gastrointestinal tract comprise a heterogeneous group of slow-growing malignancies with great differences regarding their localization, tissue of origin and their entopic and ectopic production of hormones. They can be subdivided in carcinoid tumors and endocrine tumors of the pancreas. According to their secreted products they manifest as endocrinological syndromes or as local space-occupying tumors. This review focuses, besides summarizing the available epidemiological data and describing tumor localization and classification, on the differing symptom complexes and the prognosis of the various tumor entities. Furthermore, the value of available diagnostic techniques and the role of different therapeutic modalities like surgery, radiation, biotherapy and cytostatic chemotherapy are discussed. PMID- 15779612 TI - [MR-diagnostig of arthrosclerosis]. PMID- 15779613 TI - [Epigastric pain and abnormal ECG]. AB - We report on a 50-year old man who presented with upper abdominal pain and new T wave inversions in the anterior leads of the electrocardiogramm. Since the patient additionally had cardiovascular risk factors (positive family history, smoking, hypercholesterinemia and hypertension) he was initially thought to have an acute coronary syndrome. Moreover, he had elevated serum levels of amylase and lipase, and together with typical findings of abdominal ultrasound he was diagnosed with acute pancreatitis responsible for the patient's pain. The ECG changes persisted during the hospital stay, but had fully normalised three weeks later. Further cardiologic evaluation yielded normal results. Thus, the ECG abnormalities could be attributed to the acute pancreatitis, which can cause ECG pathologies mimicking an acute coronary syndrome in up to 30%. Knowledge of there findings is important in order to prevent wrong therapeutic decisions. PMID- 15779614 TI - [Medicinal plant with heavenly power]. PMID- 15779615 TI - Hospital links patient satisfaction to employee performance. PMID- 15779616 TI - Glucose control reduces ICU stay and mortality. AB - Intensive monitoring and control of blood glucose levels in intensive care unit patients has resulted in significant clinical improvements at The Stamford Hospital in Stamford, CT. Mortality for ICU patients is down 29%, while the length of stay in the ICU has decreased by nearly 11%. PMID- 15779617 TI - Establishing criteria for physicians lowers hip fracture mortality. AB - Establishing criteria for physicians lowers hip fracture mortality. Hundreds of physicians with privileges at Staten Island University Hospital were told that they no longer could assess and treat patients who had suffered a hip fracture. It may sound like a bitter pill for the doctors to swallow, but the decision turned out to be the right one for the 785-bed hospital, located in the New York City borough of Staten Island. PMID- 15779618 TI - Florida hospital saves 5.3 M dollars by adopting principles of lean manufacturing. AB - Florida hospital saves 5.3M dollars by adopting principles of lean manufacturing. Surgery cancellations have been slashed from 13% of cases to less than 3%, while emergency department admissions have increased by 20%. Those are just two of the results of a quality improvement project at Lee Memorial Health System in Fort Myers, FL, that focused on using the principles of lean manufacturing to improve patient flow in the five-hospital system PMID- 15779619 TI - Pain therapy in horses. PMID- 15779620 TI - Diagnosing cardiac disease in equine athletes: the role of stress testing. PMID- 15779621 TI - Standing oral extraction of cheek teeth in 100 horses (1998--2003). AB - REASONS FOR PERFORMING STUDY: Extraction of cheek teeth (CT) by the conventional repulsion technique requires general anaesthesia and carries a high rate of post operative complications. Consequently, an alternative method of extraction, i.e. orally in standing horses, was evaluated. HYPOTHESIS: The need for and risks of general anaesthesia could be avoided and post extraction sequelae reduced by performing extractions orally in standing horses. METHODS: One hundred mainly younger horses (median age 8, range 2-18 years) with firmly attached CT that required extraction because of apical infections, displacements, diastemata, idiopathic fractures and the presence of supernumerary CT had the affected teeth (n = 111) extracted orally under standing sedation. Follow-up information was obtained for all cases, a median of 16 months later. RESULTS: Oral extraction was successful in 89 horses and unsuccessful in 11 due to damage to the CT clinical crown (n = 9) during extraction, for behavioural reasons (n = 1) and because the apex of a partly extracted CT fell back into the alveolus following sectioning (n = 1). Predispositions to extraction-related CT fractures were present in 5 of the 9 cases, i.e. advanced dental caries (n = 2) and pre-existing 'idiopathic' fractures (n = 3). The iatrogenically fractured CT were later repulsed under standing sedation (n = 3) and under general anaesthesia (n = 6). Eighty-one of the remaining 89 horses had successful oral CT extraction with no or minimal intra- or post operative complications occurring. Post operative complications in the other 8 cases included post extraction alveolar sequestration (n = 3), alveolar sequestration and localised osteomyelitis (n = 1), localised osteomyelitis (n = 1), incorporation of alveolar packing material into alveolar granulation tissue (n = 1), and nasal discharge due to continued intranasal presence of purulent food material (n = 1) and to ongoing sinusitis (n = 1). The above sequelae were treated successfully in all cases, with general anaesthesia required in just one case. Following oral extraction, significantly (P<0.001) fewer post operative problems developed in 54 horses with apically infected CT in comparison with 71 previous cases that had repulsion of apically infected CT at our clinic. CONCLUSIONS AND POTENTIAL RELEVANCE: Oral extraction of cheek teeth is a successful technique in the majority of younger horses with firmly attached CT and greatly reduces the post operative sequelae, compared with CT repulsion. Additionally, the costs and risks of general anaesthesia are avoided. Further experience and refinement in the described protocol could potentially increase the success of this procedure and also reduce the incidence of post operative sequelae. PMID- 15779622 TI - Lameness associated with foot pain: results of magnetic resonance imaging in 199 horses (January 2001--December 2003) and response to treatment. AB - REASONS FOR PERFORMING STUDY: The diagnosis of foot-related lameness often remains elusive and it can be difficult to offer rational treatment, or to predict outcome. OBJECTIVES: To describe the spectrum of injuries of the foot identified using magnetic resonance imaging (MRI), to determine their relative prevalence among MRI diagnoses and to establish the long-term results of treatment. METHODS: The MR images of horses examined from January 2001--December 2003 were reviewed. Horses were selected for MRI if the pain causing lameness was localised to the foot using perineural analgesia but any clinical, radiological or ultrasonographic abnormalities were insufficient to explain the degree of lameness. The clinical significance of lesions identified using MRI was determined with reference to the results of local analgesia, radiography, ultrasonography and nuclear scintigraphy. Follow-up information was obtained in January 2004 for horses which had been examined 6-36 months previously and the outcome classified as excellent (horse returned to full athletic function without recurrent lameness), moderate (sound, but only in light work), or poor (persistent or recurrent lameness). RESULTS: One hundred and ninety-nine horses underwent MRI examinations. Deep digital flexor (DDF) tendonitis was the most common injury (59%) with primary injury in 65 horses (33%) and a further 27 horses (14%) having lesions of the DDF tendon and navicular bone. Seventeen percent of horses had injuries to multiple structures, including 24 with DDF tendonitis. Desmitis of a collateral ligament (CL) of the distal interphalangeal (DIP) joint was the second most common injury (62 horses, 31%), with primary injuries in 30 horses (15%) and a further 32 horses (16%) that had CL desmitis in conjunction with other injuries. Prognosis was best for horses with traumatic injuries of the middle or distal phalanges, with 5 of 7 horses (71%) having an excellent outcome. Horses with primary lesions of the DDF tendon or CL of the DIP joint had excellent results in only 13 of 47 (28%) and 5 of 17 horses (29%), respectively. Horses with combined injuries of the DDF tendon and navicular bone, or primary navicular bone abnormalities, had a poor outcome, with the majority of horses suffering persistent lameness. CONCLUSIONS: A wide variety of lesions associated with foot pain were identified using MRI, a high proportion of which were primary soft tissue injuries with a guarded prognosis for return to full athletic function. POTENTIAL RELEVANCE: It is now possible to propose more rational treatment strategies for the variety of foot injuries identified using MRI than had previously been possible; however, further information concerning aetiopathogenesis of these injuries is needed to improve their management. PMID- 15779623 TI - Effect of systemic lidocaine on visceral and somatic nociception in conscious horses. AB - REASONS FOR PERFORMING STUDY: Commonly used analgesics (nonsteroidal anti inflammatory agents, opioids and alpha2-agonists) have unwanted side effects. An effective alternative with minimal adverse effects would benefit clinical equine pain management. OBJECTIVES: To compare the effect of lidocaine or saline on duodenal and rectal distension threshold pressure and somatic thermal threshold in conscious mature horses. HYPOTHESIS: Systemically administered lidocaine would increase somatic and visceral nociceptive thresholds. METHODS: Lidocaine (2 mg/kg bwt bolus followed by 50 microg/kg bwt/min for 2 h) or saline was administered to 6 horses each carrying a permanently implanted gastric cannula, in a randomised, blinded cross-over design. Thermal threshold was measured using a probe containing a heater element placed over the withers which supplied heat until the horse responded. A barostatically controlled intraduodenal balloon was distended until a discomfort response was obtained. A rectal balloon was inflated until extruded or signs of discomfort noted. RESULTS: Thermal threshold was increased significantly 30 and 90 mins after the start of lidocaine infusion. There was no change in duodenal distension pressure and a small but clinically insignificant change in colorectal distension pressure in the lidocaine group. CONCLUSIONS: At the dose used, systemically administered lidocaine produced thermal antinociception but minimal changes in visceral nociception. POTENTIAL RELEVANCE: At these doses, lidocaine may play a role in somatic analgesia in horses. PMID- 15779624 TI - Atropine reduces dobutamine-induced side effects in ponies undergoing a pharmacological stress protocol. AB - REASONS FOR PERFORMING STUDY: High-dose dobutamine stress echocardiography has been shown to be cardiotoxic and arrhythmogenic in horses. However, the test may have benefit in practice as a pharmacological challenge of exercise without the treadmill being required. OBJECTIVES: To investigate the effect of low-dose dobutamine on cardiac performance in ponies previously treated with atropine, in order to develop a pharmacological protocol that allows examination of the equine heart under stimulation. METHODS: In 13 healthy Shetland ponies, heart rate (HR), stroke index (SI) and cardiac index (CI) were calculated from pulsed-wave Doppler ultrasound measurements performed at rest and during incremental steps of dobutamine infusion. Group 1 (n = 7) received dobutamine infusion at 2 microg/kg bwt/min for 5 mins followed by incremental rates of 5 microg/kg bwt/min every 5 mins, from 5 to 40 microg/kg bwt/min. Group 2 (n = 6) received dobutamine infusion in incremental rates of 1 microg/kg bwt/min, every 5 mins, from 2 microg/kg bwt/min to 5 microg/kg bwt/min, after premedication with 2 injections of 25 microg/kg bwt of atropine 5 mins apart. RESULTS: The increase in CI during the pharmacological challenge was higher in Group 2 and reached about 2.5 times the resting value. This increase in CI was mediated by a significant increase in HR in both groups, while SI significantly decreased in Group 1 and did not change significantly in Group 2. Ponies of Group 1, but not those of Group 2, showed excessive restlessness and cardiac arrhythmias during the pharmacological challenge and a high intragroup variability in cardiac response. CONCLUSIONS: The results of this study suggest that a low dose of dobutamine in ponies previously given atropine could be a helpful pharmacological protocol to perform stress echocardiography in equids. POTENTIAL RELEVANCE: Further studies should evaluate left ventricular wall motion in horses undergoing low-dose dobutamine protocol after pretreatment with atropine. PMID- 15779625 TI - In vitro comparison of three materials as apical sealants of equine premolar and molar teeth. AB - REASONS FOR PERFORMING STUDY: Surgical endodontic therapy is a conservative dental technique used in horses with some degree of clinical success. Failure of this procedure can partially be explained by inadequate sealing of the root apices with resultant microleakage in the periapical area. OBJECTIVES: To assess and compare in vitro sealing ability of 3 different dental restorative materials used as apical sealants during equine surgical endodontics. METHODS: Thirty extracted equine cheek teeth were divided randomly into 3 groups and subjected to apicoectomy and apical sealing using 3 materials: reinforced zinc oxide-eugenol cement; intermediate restorative material (IRM); a resin-modified glass ionomer; and amalgam. After apical sealing, the teeth were submerged in a solution of Procion Brilliant Cresyl Blue stain for a period of 7 days. The teeth were then washed, embedded in resin, sectioned and assessed microscopically for dye leakage around the apical restorations. RESULTS: Although the materials proved effective as apical sealants, some dye leakage was encountered in all 3 groups with no statistical difference (P = 0.114). CONCLUSIONS AND POTENTIAL RELEVANCE: IRM, a resin-modified glass ionomer and amalgam all showed comparative features as apical sealants when used in vitro in equine teeth. IRM is currently regarded as the superior material in clinical situations due to its ease of handling and lesser sensitivity to environmental moisture during placement compared to the other 2 materials. PMID- 15779626 TI - Modified frontonasal sinus flap surgery in standing horses: surgical findings and outcomes of 60 cases. AB - REASONS FOR PERFORMING STUDY: Previous studies of sinus surgery on standing horses have been based on techniques which utilise powered hand bone saws and preserve the bone flap, which was thought to be essential for a good cosmetic result. This report describes a simplified technique applied to the standing horse where the sinus flap construction used a large skull trephine and where the bone flap is necessarily discarded. OBJECTIVES: To assess whether the modified standing frontonasal flap (SFF) surgery offers an effective method to investigate and/or treat sinunasal disorders in the horse. METHODS: The case records of 60 horses subjected to modified SFF surgery were reviewed to analyse the efficacy of the technique when applied to a range of sinunasal disorders in terms of diagnosis, surgical findings, complications and long-term outcomes, including cosmetic effect. RESULTS: Resolution of clinical signs was achieved in 54 out of 60 horses, and for the remainder the SFF technique proved useful on a diagnostic basis. The cosmetic result was, in the opinion of the owners, excellent or satisfactory in 48 of 56 (86%) horses. CONCLUSIONS: The modified SFF technique was found to be practicable and provided satisfactory exposure of the sinus contents for diagnosis and removal of diseased tissue in a range of disorders. An acceptable cosmetic result can be obtained in the majority of horses despite rejection of the disc of overlying bone. Apart from avoiding the risks associated with general anaesthesia, the advantages of the modified SFF sugery are: a reduction of haemorrhage in the standing horse; reduced mucosal engorgement; straightforward anatomical orientation; a comfortable working height; and reduced surgery time. A sound knowledge of the functional anatomy of the area is a prerequisite before embarking on any form of sinunasal surgery. Complications are infrequent and usually transient. POTENTIAL RELEVANCE: The modified SFF technique is a valuable alternative to other invasive methods of sinunasal surgery. PMID- 15779627 TI - Genetic diagnosis of sex chromosome aberrations in horses based on parentage test by microsatellite DNA and analysis of X- and Y-linked markers. AB - REASONS FOR PERFORMING STUDY: Sex chromosome aberrations are often associated with clinical signs that affect equine health and reproduction. However, abnormal manifestation with sex chromosome aberration usually appears at maturity and potential disorders may be suspected infrequently. A reliable survey at an early stage is therefore required. OBJECTIVES: To detect and characterise sex chromosome aberrations in newborn foals by the parentage test and analysis using X- and Y-linked markers. METHODS: We conducted a genetic diagnosis combined with a parentage test by microsatellite DNA and analysis of X- and Y-linked genetic markers in newborn light-breed foals (n = 17, 471). The minimum incidence of sex chromosome aberration in horses was estimated in the context of available population data. RESULTS: Eighteen cases with aberrations involving 63,XO, 65,XXY and 65,XXX were found. The XO, XXY (pure 65,XXY and/or mosaics/chimaeras) and XXX were found in 0.15, 0.02 and 0.01% of the population, respectively, based solely on detection of abnormal segregation of a single X chromosome marker, LEX003. CONCLUSIONS AND POTENTIAL RELEVANCE: Detection at an early age and understanding of the prevalence of sex chromosome aberrations should assist in the diagnosis and managment of horses kept for breeding. Further, the parental origin of the X chromosome of each disorder could be proved by the results of genetic analysis, thereby contributing to cytogenetic characterisation. PMID- 15779628 TI - Functional adaptation of articular cartilage from birth to maturity under the influence of loading: a biomechanical analysis. AB - REASONS FOR PERFORMING STUDY: The concept of functional adapatation of articular cartilage during maturation has emerged from earlier biochemical research. However, articular cartilage has principally a biomechanical function governed by joint loading. OBJECTIVES: To verify whether the concept of functional adaptation can be confirmed by direct measurement of biomechanical properties of cartilage. HYPOTHESIS: Fetuses have homogeneous (i.e. site-independent) cartilage with regard to biomechanical properties. During growth and development to maturity, the biomechanical characteristics adapt according to functional (loading) demands, leading to distinct, site-dependent biomechanical heterogeneity of articular cartilage. METHODS: Osteochondral plugs were drilled out of the surface at 2 differently loaded sites (Site 1: intermittent impact-loading during locomotion, Site 2: low-level constant loading during weightbearing) of the proximal articular cartilage surface of the proximal phalanx in the forelimb from stillborn foals (n = 8), horses of age 5 (n = 9) and 18 months (n = 9) and mature horses (n = 13). Cartilage thickness was measured using ultrasonic, optical and needle-probe techniques. The osteochondral samples were biomechanically tested in indentation geometry. Young's modulus at equilibrium, dynamic modulus at 1 Hz and the ratios of these moduli values between Sites 1 and 2 were calculated. Age and site effects were evaluated statistically using ANOVA tests. The level of significance was set at P<0.05. RESULTS: Fetal cartilage was significantly thicker compared to the other ages with no further age-dependent differences in cartilage thickness from age 5 months onwards. Young's modulus stayed constant at Site 1, whereas at Site 2 there was a gradual, statistically significant increase in modulus during maturation. Values of dynamic modulus at both Sites 1 and 2 were significantly higher in the fetus and decreased after birth. Values for both moduli were significantly different between Sites 1 and 2 from age 18 months onwards. The ratio of values between Sites 1 and 2 for Young's modulus and dynamic modulus showed a gradual decrease from approximately 1.0 at birth to 0.5 0.6 in the mature horse. At age 18 months, all values were comparable to those in the mature horse. CONCLUSIONS: In line with the concept of functional adaptation, the neonate is born with biomechanically 'blank' or homogeneous cartilage. Functional adaptation of biomechanical properties takes place early in life, resulting in cartilage with a distinct heterogeneity in functional characteristics. At age 18 months, functional adaptation, as assessed by the biomechanical characteristics, has progressed to a level comparable to the mature horse and, after this age, no major adaptations seem to occur. POTENTIAL RELEVANCE: Throughout life, different areas of articular cartilage are subjected to different types of loading. Differences in loading can adequately be met only when the tissue is biomechanically adapted to withstand these different loading conditions without injury. This process of functional adaptation starts immediately after birth and is completed well before maturity. This makes the factor of loading at a young age a crucial variable, and emphasises the necessity to optimise joint loading during early life in order to create an optimal biomechanical quality of articular cartilage, which may well turn out to be the best prevention for joint injury later in life. PMID- 15779629 TI - Dose responses to inhalation of endotoxin, hay dust suspension and Aspergillus fumigatus extract in horses as measured by levels and activation of matrix metalloproteinase-9. AB - REASONS FOR PERFORMING STUDY: Airway matrix metalloproteinases (MMPs) increase after endotoxin (LPS) exposure, but there are no reports describing dose dependent increases or activation following exposure. OBJECTIVES: To study matrix metalloproteinase-9 (MMP-9) and -2 (MMP-2) responses in bronchoalveolar lavage fluid (BALF) from heaves-susceptible and control horses following inhalation of hay dust suspension (HDS), LPS and Aspergillus fumigatus extract (AFE). METHODS: Heaves-susceptible (n = 7) and control (n = 6) horses received inhalation challenges with 3 different doses of HDS and LPS. Heaves-susceptible horses (n = 6) also received 3 different doses of AFE and one dose of AFE depleted of endotoxin (AFE-LPS). BALF collected following inhalation challenges was analysed using gelatin zymography. Gelatinolytic bands were identified as complex, pro-MMP 9, active MMP-9, pro-MMP-2 and active MMP-2 based on molecular weights. RESULTS: Each challenge substance induced a dose-dependent elevation in gelatinolytic activity. The dose-dependency was most evident for pro-MMP-9 and total MMP-9 levels in heaves-susceptible horses following LPS challenges. CONCLUSIONS: There is a dose-dependent elevation in MMP-9 in BALF of heaves-susceptible and control horses following inhalation challenge with organic dust and some of its components, elevation being more marked in heaves-susceptible horses. Organic dust components vary in their pro-inflammatory potential. POTENTIAL RELEVANCE: This study supports the role of MMPs in the pathogenesis of heaves and highlights the potential value of protease inhibitors in attenuating the airway inflammatory response to inhaled organic dust. PMID- 15779630 TI - Deep digital flexor tendon force and digital mechanics in normal ponies and ponies with rotation of the distal phalanx as a sequel to laminitis. AB - REASONS FOR PERFORMING STUDY: Previous studies have implicated tension in the deep digital flexor tendon (DDFT) in the rotation of the distal phalanx (DP) after the breakdown of the dorsal laminae caused by laminitis. Howeveer, once the DP has rotated, the DDFT should become shorter, reducing the force it exerts on the DP. OBJECTIVE: To compare DDFT force and ground reaction forces (GRFs) in normal ponies and ponies with rotation of the DP as a sequel to laminitis. METHODS: Six normal ponies (Group 1) and 6 sound ponies with 6-13 degrees of rotation of the DP in relation to the dorsal hoof wall (Group 2) were assessed at trot using forceplate and motion analysis. The force in the DDFT was calculated by assuming that the extending moment at the distal interphalangeal (DIP) joint resulting from the GRF was equal to the flexing moment created by the force in the DDFT during the stance phase (inverse dynamics). RESULTS: In early stance, the peak DDFT force (mean+/-s.d.) in the normal ponies was 1.92+/-1.63 N/kg. However, in Group 2, the point of zero moment was palmar to the centre of rotation of the DIP joint for the first 40% of stance and hence DDFT force was zero. Force in the DDFT reached a peak of 10.00+/-3.56 N/kg at 60.7+/-5.6% of stance in Group 1 and 6.41+/-1.37 N/kg at 79.2+/-9.6% of stance in Group 2. CONCLUSIONS: DDFT force in Group 2 laminitic ponies was much reduced until late stance, when it neared normal values. POTENTIAL RELEVANCE: Further studies of ponies with rotation of the DP as a sequel to laminitis should assist farriery aimed at reducing the force in the DDFT through the breakover phase of stance to protect damaged dorsal laminae. PMID- 15779631 TI - Osteoarthritis of the talocalcaneal joint in 18 horses. AB - REASONS FOR PERFORMING STUDY: Talocalcaneal osteoarthritis (TO) is an uncommon cause of moderate to severe hindlimb lameness, on which only isolated case reports have been published to date. OBJECTIVES: To review the clinical features of TO and determine optimal methods for diagnosis, management and prognosis. METHODS: The case records from 4 referral centres of 18 horses showing hindlimb lameness considered, as a result of clinical investigation, to be caused by TO, were reviewed. RESULTS: TO affected mature sports and pleasure horses (age 7-16 years) and caused moderate to severe lameness, usually of sudden onset with no obvious inciting cause. There were few localising signs, other than worsening of lameness by hock flexion. Tarsocrural joint analgesia produced improvement in lameness in 6/11 horses (55%) and perineural analgesia of the tibial and fibular nerves complete soundness in 6/14 horses (43%) in which it was performed; 7/14 horses (50%) showed a further substantial improvement. Radiological findings included subchondral bone lysis and sclerosis and irregular joint space width, seen most obviously in a lateromedial view. Nuclear scintigraphy revealed marked uptake of radiopharmaceutical predominantly plantaromedially in the region of the talus in the 7 horses in which it was performed. Fourteen horses were treated conservatively with box- or pasture-rest, with or without intra-articular corticosteroids, hyaluronic acid or polysulphated glycosaminoglycan, and all remained lame. Intra-articular corticosteroids appeared to have no effect in any horse. Of 10 horses receiving conservative management only, 6 were subjected to euthanasia, 3 were retired and 1 remained in light work, but was still lame. Two horses treated by either partial tibial and fibular neurectomy or subchondral forage failed to regain soundness and were retired. Six horses were treated by surgical arthrodesis of the talocalcaneal joint with 2 or three 5.5 mm AO screws introduced obliquely across the joint from the plantarolateral aspect of the calcaneus, which resulted in improvement in lameness in all cases. CONCLUSIONS: Osteoarthritis of the talocalcaneal joint causes acute onset severe lameness, but clinical findings and diagnostic analgesia often fail to identify precisely the site of pain. Consistent radiographic changes suggested TO was contributing to the lameness and this diagnosis was supported by nuclear scintigraphy. The poor success of conservative treatment (including intra-articular medication) suggests that surgical arthrodesis is the treatment of choice, although the prognosis is still poor for a return to full soundness. POTENTIAL RELEVANCE: The clinical features described should facilitate more accurate diagnosis and prognosis. A novel surgical treatment is described which appears to offer significant improvement in the lameness. Further work is necessary to determine the causes of this condition and more effective management. PMID- 15779632 TI - Concentration of methylprednisolone in the centrodistal joint after administration of methylprednisolone acetate in the tarsometatarsal joint. AB - REASONS FOR PERFORMING STUDY: The centrodistal (CD) and tarsometatarsal (TMT) joints are often injected individually with a corticosteroid to resolve lameness caused by osteoarthritis (OA). There are no data available regarding diffusion of methylprednisolone (MP) from the TMT joint to the CD joint. HYPOTHESIS: A therapeutic concentration of MP diffuses into the CD joint after methylprednisolone acetate (MPA) is administered into the TMT joint. OBJECTIVE: To measure the concentration of MP in the CD joint after MPA was administered into the TMT joint. METHODS: MPA was administered into a TMT joint of 16 horses. At different times, the ipsilateral CD joint of these horses was injected with a small amount of saline and recovered saline was measured for concentration of MP using high performance liquid chromatography. RESULTS: Six hours after administration of MPA into the TMT joint, a therapeutic concentration of MP was found in all 10 CD joints sampled at this time. CONCLUSIONS: Horses with pain arising from the distal 2 joints of the hock can be treated by administering MPA into the TMT joint alone. POTENTIAL RELEVANCE: Administering MPA into the TMT joint only, to treat OA of the distal 2 hock joints, reduces the difficulties and risks associated with centesis of the CD joint. PMID- 15779634 TI - Clinical diagnosis and results of surgical treatment of 13 cases of acquired bilateral stringhalt (1991--2003). PMID- 15779633 TI - Effect of age at presentation on outcome following arthroscopic debridement of subchondral cystic lesions of the medial femoral condyle: 85 horses (1993--2003). AB - REASONS FOR PERFORMING STUDY: Subchondral cystic lesions of the medial femoral condyle (SCMFC) are well documented in horses < or =3 years; arthroscopic debridement or enucleation of the cyst is currently the surgical treatment of choice. However, studies of occurence and outcome following surgery in older horses are lacking. OBJECTIVE: To identify factors important in outcome for horses with SCMFC treated by arthroscopic debridement. HYPOTHESIS: Age of horse at presentation has a significant influence on return to work following arthroscopic treatment for SCMFC. Clinical and diagnostic findings are also significant with respect to prognosis. METHODS: A retrospective review of medical records from 6 equine referral centres identified 85 horses that underwent arthroscopic debridement of SCMFC. Clinical examination, radiographic and arthroscopic findings were analysed together with follow-up data. Univariable analysis and multivariable logistic regression models were used to determine factors affecting return to soundness. Event-time analysis was performed to evaluate return to work. RESULTS: Older horses (>3 years) were less likely to return to soundness (P = 0.02) or to work (P = 0.04) than younger horses (< or = 3 years). Of 39 horses age 0-3 years, 25 (64%, 95% CI 49-79%) returned to soundness. Of 46 horses age >3 years, 16 (35%,95% CI 21-49%) returned to soundness. In addition, cartilage damage at sites other than the SCMFC negatively affected prognosis (P = 0.05). The hospital where treatment was performed had no influence on return to soundness. CONCLUSIONS: Older horses carry a worse prognosis for both return to soundness and return to work. POTENTIAL RELEVANCE: It is important for clients to be made aware of the difference in outcome between age groups. PMID- 15779635 TI - Let's tackle errors and omissions head-on. PMID- 15779637 TI - A critical review of the aetiology of diabetic neuropathic ulcers. AB - Much has been written on the causes of diabetic neuropathic foot ulcers. This review outlines current thinking on the main aetiologies and describes the causative factors involved. Such an understanding will aid patient assessment. PMID- 15779636 TI - Making the case for adapting the abbreviated burn severity index to include erythroblast count. AB - The presence of erythroblasts is an accurate predictor of a poor prognosis. This study investigates whether this should be taken into account when assessing patients with burn injuries as traditional protocols may underestimate the level of risk. PMID- 15779638 TI - The positive experience of using a growth factor product on deep wounds with exposed bone. AB - Trafermin, a form of basic fibroblast growth factor, has been used in Japan since 2001. This study investigates whether it can facilitate closure in wounds with deep soft-tissue defects and exposed bone, where surgical closure is not possible. PMID- 15779639 TI - Treatment of dehisced and infected wounds. PMID- 15779640 TI - An unusual night at a burns unit. PMID- 15779642 TI - The effectiveness of a hyperoxygenated fatty acid compound in preventing pressure ulcers. AB - OBJECTIVE: To compare the effects of Mepentol, a hyperoxygenated fatty acid preparation, with a placebo treatment in preventing the development of pressure ulcers. METHOD: The research study consisted of a multicentre double-blind randomised clinical trial. The incidence of pressure ulcers, relative risk (RR), preventable fraction and number necessary to treat (NNT) were calculated. In addition, Kaplan-Meier survival curves, with log-rank test, and Cox's proportional hazards regression model were used to compare both groups. RESULTS: A total of 331 patients completed the study: 167 in the control group and 164 in the study group. Pressure-ulcer incidence during the study was 7.32% in the intervention group versus 17.37% in the placebo group (p 0.006). These results show that for each 10 patients treated with Mepentol one pressure ulcer was prevented (NNT = 9.95). Survival curves and the regression model showed a significant statistical difference for both groups (p < or = 0.001). The average cost of Mepentol during the study was 7.74 Euro. CONCLUSION: Mepentol is an effective measure for pressure ulcer prevention. It was more effective than a greasy placebo product, and was found to be cost-effective. PMID- 15779641 TI - Pressure ulcers and spinal-cord injury: incidence among admissions to the Irish national specialist unit. AB - Unlike many other countries, Ireland does not have a pathway to facilitate rapid transfer to its national spinal injuries unit. This retrospective study investigated the effect of this on the incidence of pressure ulceration in new admissions to the unit. PMID- 15779643 TI - A review of studies that examine the impact of infection on the normal wound healing process. AB - Wound infection disrupts the normal healing process, although to what extent is not proven. This review explores controversies in the literature surrounding the bacterial load of the chronic wound and its impact, if any, on this process. PMID- 15779644 TI - Measuring quality of life in patients with pressure ulcers to include in economic evaluations. AB - Quality-of-life measures are increasingly being incorporated into economic evaluations of health-care interventions. But this review identified only two studies with empirical data on quality of life in patients with pressure ulceration. PMID- 15779645 TI - Effects of warming on healing. AB - Systemic and local warming may accelerate wound healing and minimise postoperative wound infection, although more research is needed to confirm this. The existing evidence of the benefits of warming therapies is outlined here. PMID- 15779646 TI - [Methodology of recommendations]. PMID- 15779647 TI - [TNM 2002 classification of urogenital tumors: principles and new findings]. PMID- 15779648 TI - [Presentation of a urologic cancer file in team work]. PMID- 15779649 TI - [Information communication and personalized care program]. PMID- 15779650 TI - [Standardized histopathology reports in urologic oncology]. PMID- 15779651 TI - [Comorbidity and urologic cancer]. PMID- 15779652 TI - [Testicular tumors]. PMID- 15779653 TI - [Penile tumors]. PMID- 15779654 TI - [Prostatic cancer]. PMID- 15779655 TI - [Urothelial tumors]. PMID- 15779656 TI - [Kidney tumors]. PMID- 15779657 TI - [Adrenal gland tumors]. PMID- 15779658 TI - Leadership building partnerships. PMID- 15779659 TI - Oral care in intensive care units: a literature review. AB - Patients in an intensive care unit (ICU) may have significant oral health needs. This article offers a review of the available literature regarding oral care for patients in the ICU. Based on this review, the author describes signs and symptoms reported as common among patients in the ICU and makes recommendations concerning their oral care. The implications of these findings are also discussed. PMID- 15779660 TI - Non-clinical information obtained by dentists during initial examinations of older adult patients. AB - The authors sent a questionnaire to a random sample of general dentists in Ontario, Canada, to assess the types of non-clinical information (NCI) dentists usually obtain during initial examinations of older patients. From a list of 11 NCI questions, dentists indicated which questions they usually asked during new patient examinations. The adjusted response rate was 34% (n = 672). Respondents most often asked about pain and satisfaction with the appearance of teeth and/or dentures. About half the respondents asked about oral dryness and whether problems with chewing had limited food choices. Respondents were least likely to ask about problems with speaking and avoidance of eating with others because of chewing problems. Traits of those who asked the least common NCI questions typically including continuing education courses in geriatric dentistry, self perceived competence in treating elderly adults living in institutional settings, exposure to geriatric outreach settings during dental school and greater dentist involvement in patient history taking. PMID- 15779661 TI - Dental hygienists as providers in long-term care facilities. AB - The authors surveyed all practicing dental hygienists (n = 870, response rate = 69%) in Kansas about providing preventive dental care to elderly residents of long-term care (LTC) facilities. Maximum likelihood estimates stepwise logistic regressions were used to predict the likelihood of dental hygienists filling the need for a specialty workforce. Few respondents wanted to work exclusively with elderly patients. Lack of interest in LTC practice was related to the number of children living at home or discomfort with patients who had limited abilities to communicate. Interest in part-time geriatric practice was related to motivation to help older patients and unhappiness with current role limitations. Implications for reimbursement and educational offerings are discussed. While dental hygienists could fill a need in preventive oral health for elderly patients, it remains to be seen if subsequent policy changes result in that outcome. PMID- 15779662 TI - Factors associated with self-reported use of dental health services among older Greek and Italian immigrants. AB - The authors discuss utilization of dental health services by older Greek and Italian immigrants in Melbourne, Australia. Their study involved 374 Greek and 360 Italian adults who completed a questionnaire and received an oral examination. Nearly 41% of Greek and 45% of Italian respondents had used dental services in the previous year. As barriers to care, Greek participants most often cited waiting lists and waiting time in the office. Italian participants most often identified cost, length of waiting lists and language barriers. Multivariate analyses associated recent use of dental services with number of teeth, oral health knowledge, age and occupation before retirement for both groups, as well as living arrangements among Greek participants and perceived barriers among Italian participants. Findings highlight the need for oral health promotion programs targeted toward older adults from immigrant populations and reductions of the structural barriers that prevent these adults from seeking oral health care. PMID- 15779663 TI - Arbitrarily primed polymerase chain reaction for fingerprinting the genotype identification of mutans streptococci in children with Down syndrome. AB - This study investigated the possible intrafamilial similarity of mutans streptococcal strains in some families with a child with Down syndrome using chromosomal DNA fingerprinting. The isolates were genotyped using arbitrarily primed polymerase chain reaction with the OPA 02 and OPA 03 primers. The results showed that five children with Down syndrome harbored mutans streptococci genotypes different from those of their mothers. A matching of genotypes was observed within the control pair (mother/child without Down syndrome). After six months, new samples were collected from all participants. Analysis showed that samples from children with Down syndrome were colonized by a new strain of Streptococcus mutans that did not match the previously collected one. The results suggest the S. mutans indigenous bacteria change more than once in children with Down syndrome. PMID- 15779664 TI - [Functional disturbances of the hypothalamus in patients with anorexia nervosa]. AB - Anorexia nervosa is a complex disease characterized by abnormal feeding behaviour, food aversion and acute disturbances in perception of the body shape. Every eating disorder comes about as a consequence of disturbances in synaptic transmission in particular brain regions (hypothalamus, limbic system, cortical centres). The human and animal feeding physiology is precisely regulated by autonomic nuclei of hypothalamus. The perikarya of arcuate nucleus, lateral hypothalamic nuclei and other areas produce and release both the orexigenic (e.g. NPY) and anorexigenic (e.g. a-MSH) signalling substances. The novel hypothalamic and peripheral neurohormones: orexins and ghrelin as well as serotonin and dopamine seem to play a significant role in pathogenesis of eating disorders. In anorexia nervosa simultaneous excess of orexigenic and anorexigenic factors may evoke a "mixed signal" leading to failure of hypothalamic regulatory pathways. Experimental results also suggest that women with anorexia nervosa have disturbances of regional cerebral blood flow. PMID- 15779665 TI - [Characteristics of the forming personality in children suffering from anorexia nervosa]. AB - AIM: The evaluation of forming personality in 30 children (27 girls and 3 boys) suffering from anorexia nervosa (average age--13.5). METHOD: Anamnesis from patients and their parents, clinical observation, psychiatric investigation, psychological investigation with use of following methods: The Thematic Apperception Test, Colorful Pyramids Test, The House-The Tree-The Man Test. RESULTS: After the conducted investigation and observation, the characteristics of the forming incorrect personality were observed in 80% of examined children. According to DSM IV classification, in 50%--from the cluster C, and in 30% cases- the cluster B. According to ICD-10 classification: 26.6% attributes of dependent personality, 16.6%--histrionic personality, 13.3% avoiding personality, 10%- anancastic personality, 6.7% borderline personality and 6.7% antisocial personality. CONCLUSIONS: There is predominating opinion in literature, connecting restrictive type of psychical anorexia, with disorders of personality of anxiety type--the cluster C, and the figure bulimic--with personality the cluster B. Those results were confirmed with statistical significance in the conducted examinations. PMID- 15779666 TI - [The self-image and the image of other people in the perception of patients with anorexia nervosa]. AB - AIM: The aim of this study was to compare the self-image and the image of other people in the perception of female patients with anorexia nervosa. METHOD: Thirty women with anorexia nervosa were investigated during hospitalisation in the Psychiatric Department of the Medical School of Lublin. The mean age of patients was 20 years. The Adjective Check List (ACL) of Gough and Heilbrunn was used with the instruction: how you are, how would you like to be, how are the other people. RESULTS: The measurement gave us the possibility to obtain the real and ideal self-image and the real image of the other people. The results showed that there are statistically significant differences between self-image and the image of other people in the perception of anorectic females in connection with self acceptance, self-control, the need of intraception, nurturance, exhibition, autonomy, aggression, need of abasement, difference, the scale of masculine attributes, the scale of feminine attributes and the scale of the free child. CONCLUSIONS: Significant differences occurred also in the scale based on the results. The final conclusions are as follows: 1) Anorectic females show negative self-image and even a more negative image of other people. 2) According to anorectic females the people are more aggressive, more self-issued, avoiding close contacts, manipulating, tending to gain autonomy and to be in the centre of the group's attention. PMID- 15779667 TI - [The self-acceptance and the acceptance of the parents by patients with anorexia nervosa]. AB - AIM: The aim of this work is to describe the relationship between self-esteem and characteristics of self image and acceptance of parents and the characteristics of their images in the perception of patients with anorexia nervosa. METHOD: The test group consisted of 30 patients diagnosed with anorexia nervosa undergoing a treatment in the Department of Psychiatry of the Medical Academy in Lublin. The average age of the examined patients was 20 years. All patients had secondary education. In my work the ACL Gough and Heilbrun Adjectival Test was used which was completed by the examined women three times following the instruction: "I am", "my mother is", "my father is". As a result the actual images of the patients as well as of their mothers and fathers were obtained. The esteem rates were calculated by comparing the number of positive adjectives to the number of negative adjectives used by the patients to evaluate individual persons. The interdependencies between esteem rates and the characteristics of the actual images of these persons were determined based on the r-Pearson correlation. RESULTS AND CONCLUSIONS: The results obtained indicated the existence of crucial statistic dependencies between the variables analysed and constituted the basis for the formulation of the following conclusions: 1. Patients are characterised by a low self-esteem and a low acceptance level of their parents. They accept their fathers in the smallest degree, compared to that their self-esteem is higher and the acceptance of their mothers is the highest. 2. Self-esteem is related in a significant way to the characteristics of self image such as: sense of responsibility, autonomy, ability to accomplish life objectives and establishing close relationships with other people. 3. The acceptance of their mothers is related to the characteristics typical of them such as: protectiveness, ability to establish and maintain positive interpersonal relationships, empathy and understanding the motives of their own behaviour and those of other people. 4. The acceptance of their fathers is related in a significant way to such characteristics discerned in them as: perseverance, ability to work hard, protectiveness, responsibility and ability to cope with tasks and problems and establishing close relationships with other people. PMID- 15779668 TI - [Temperament and character of Polish women with anorexia and bulimia nervosa]. AB - AIM: One of the factors influencing eating disorders are personality traits. The authors analyse temperament and character of healthy women. METHOD: The Cloninger Temperament and character Inventory was applied to 52 eating disordered patients (33 with anorexia nervosa and 19 with bulimia nervosa). The patients were divided into subgroups of restrictive type and bulimic types of anorexia, bulimia and bulimic episodes. RESULTS: In all the subgroups of the patients a higher result was obtained on the harm avoidance scale (HA), cooperativeness (C) and the self transcendence ST2 subscale. Lower results were seen in self-directedness (SD) in the SD2, SD3 and SD5 subscales. The subgroups differed in temperament. Bulimia patients noted a higher need for NS stimulation and a higher reward dependence (RD). Anorectic patients had higher results in the persistance scale (P), whilst the restrictive anorectic patients had lower results in the NS1 and RD3 subscales. CONCLUSIONS: The TCI Inventory is a useful tool, helping for a precise measurement of the difference in temperament of anorectic and bulimia patients as compared to their healthy peers. PMID- 15779669 TI - [An attempt to evaluate the health condition and social functioning of anorectic females--a follow-up study]. AB - AIM: An attempt to evaluate, on the basis of follow-up data, the health condition and social functioning in patients with anorexia nervosa, four years after hospitalisation. METHOD: Anamnesis from 20 young females (aged 17-24, average 21.7) hospitalised for the treatment of anorexia nervosa in The Department of Developmental, Psychotic and Geriatric Psychiatry, Medical University in Gdansk. RESULTS AND CONCLUSIONS: 50% of those studied suffer protractedly, with periods of remission or intensity of symptoms. 80% of women have achieved normalisation of weight (30% of them seemed to be healthy and well functioning; 50% of them is in partial remission), 20% still fulfill DSM-IV criteria for anorexia nervosa. Currently 60% menstruate (10% of them without hormonal treatment). In the examined group, 50% appear to have an abnormal manner of nutrition. Abnormal behaviour caused weight loss in 70% of those studied. Most of the females (55%) consider themselves overweight, 75% have weight-gain phobia. PMID- 15779670 TI - [Altered visual information processing in anorectic girls--the study on potentials related to the test of attention]. AB - AIM: Combined biological and psychological basis of anorexia nervosa, related to increased self-control, low self-esteem and peculiar motivation, makes a synthetic approach still hard to achieve. The presented study assessed cognitive functions in anorectic patients with respect to both neuropsychological (test of attention) and biological (electrophysiology of information processing) approaches, and their reciprocal interrelations as well. METHODS: Eleven adolescent anorectic females and 10 control subjects performed the Continuous Attention Test CAT. Parameters of potentials elicited by the visual CAT items were referred to the CAT results. RESULTS: The anorectic girls were more accurate during CAT performance, revealing a lower index of errors, especially of commissions. On the other hand, they revealed a lower amplitude of the P2 component (correlated with index of commissions) and shortened latency of the frontal N2 component (correlated with a lower index of errors). The groups differed in relations between psychometric and biological parameters referring to the earlier stages of information processing. In the control group, high index of detections was correlated to right-sided P1 and P2 components. In the patient group, correlations linked a lowered index of commissions with right-sided N1 and bilateral P2 amplitudes. CONCLUSIONS: Increased self-control in anorexia nervosa, reflected by lower frequency of needless reactions, is related to alterations in biological visual information processing yet on its early, preconscious stage. PMID- 15779671 TI - [Assessment of early traumatic life events influence on intensity and frequency of symptoms in patients aged 50 years and over]. AB - AIM: Assessment of early traumatic life events influence on psychological symptoms of depressive syndromes. METHOD: 88 patients with diagnosis of depression were included. The inventory of psychopathological symptoms--AMDP and depression rating scales--MADRS, GDS were applied. RESULTS: Early traumatic life events were associated with general intensity of depression, but also with specific categories of symptoms (thinking disturbances, phobias, obsessions). CONCLUSIONS: Traumatic life events in early childhood may in effect lead to changes in the clinical picture of depression in adulthood. The strength of the particular events may vary. PMID- 15779672 TI - [CYP 2D6 activity profile among patients with depression]. AB - AIM: The aim of this work was to establish the profile of CYP 2D6 activity among patients with depressive disorders, and compare it with the literature data on this activity among healthy volunteers, recruited from the Polish population. METHOD: The debrisoquine test was used to determine CYP 2D6 activity. Debrisoquine metabolic ratio (MR) was determined with the use of high performance liquid chromatography with spectrophotometric detection. Our results were compared with the results obtained with the use of an analogous method in the study performed at the Institute of Cardiology in Warsaw on 154 healthy controls. RESULTS: The percentage of "ultra-fast metabolizers" was 1.03% in the group of patients with depression, and 1.3% in the group of healthy volunteers. The percentages of the "slow metabolizers" in both groups were 5.1% and 5.6% respectively. Also, the percentages of persons considered to be at an increased risk of undesirable side effects from drugs metabolized by CYP 2D6, i.e. persons with debrisoquine metabolic ratio values above 4 were nearly identical in both groups (11.3 and 11.6%). CONCLUSION: The results showed that CYP 2D6 activity profiles were practically identical in both groups. This indicates that there is no relationship between CYP 2D6 gene expression, as measured by the debrisoquine test, and affective disorders morbidity. PMID- 15779673 TI - [Assessment of CYP2D6 activity as a form of optimizing antidepressant therapy]. AB - Cytochrome 2D6 catalyzes oxidation processes of many antidepressants (TCAs, SSRIs, maprotyline, mianserine, nefazodon, trazodon, venlafaxine). CYP2D6 is characterized by genetically determined polymorphism which may lead to serious clinical consequences. Based on CYP2D6 activity four phenotypes are distinguished: poor metabolism (PM), intermediate (IM), extensive (normal) EM and ultrarapid (UM). In case of PM and IM increased plasma concentration of a drug and adverse events or toxicity may appear. Decreased plasma level and lack of clinical effect may be connected with the ultrarapid phenotype. CYP2D6 activity may be assessed by phenotyping or genotyping . Model drugs such as sparteine, debrisoquine, dextromethorphan and metoprolol are used in the phenotyping method. Based on the metabolic ratio of model drug the phenotype status is established. Genotyping consists in an assessment of genotype i.e. an identification of alleles coding the CYP2D6 protein. The environmental factors may modify the CYP2D6 activity and have influence on phenotyping but not genotyping results. The knowledge of CYP2D6 phenotype is of special value when drugs characterized by a narrow therapeutic index are used and in polymedicated and older patients. PMID- 15779675 TI - [Some aspects of pharmacotherapy in the context of transference countertransference]. AB - The article deals with the psychological background of administration of medication in psychiatry, the meaning of administration of drugs in relation to the doctor-patient relationship. The place of pharmacotherapy among other medical procedures was discussed. The differences in the approach to pharmacotherapy in different therapists was noticed, with the pointing out to two kinds of approaches (symptom oriented and problem oriented). Other than rational motivations for the use of pharmacotherapy was discussed. PMID- 15779674 TI - [Changes of sleep quality and mood disorders under the influence of phototherapy in patients with seasonal affective disorders SAD]. AB - Major depression, seasonal pattern (seasonal affective disorder SAD) characterize the winter recurrence depressive episodes with remission of symptoms in spring and summer. Patients with winter depression report hypersomnia, fatigue, loss of energy, carbohydrate craving, appetite and weight gain. AIM: The aim of this study was to assess the effect of phototherapy on the quality of sleep parameters and subjective estimation of mood disorders in patients with seasonal affective disorders. METHOD: The investigated group consisted of 17 patients with SAD (15 female, 2 male) aged 18-64 (mean 38+/-12) years. Phototherapy (bright light therapy) was applied for 14 days, everyday morning--30 minutes, between 6.00 to 10.00--exposition to light of about 10,000 lux intensity. Polysomnogram (sleep EEG) was recorded before and after treatment. RESULTS: After phototherapy patients reported a significant mood improvement (57%) measured by the Seasonal Pattern Assessment Questionnaire. Sleep investigation showed: increased sleep efficiency, decreased sleep latency, decreased slow wave sleep latency and increased of sleep spindles in the first hour of sleep. CONCLUSIONS: Research confirms that phototherapy is an effective method of treatment of choice for patients with SAD. The result indicates that phototherapy markedly improved mood and sleep quality. PMID- 15779676 TI - [Application of actigraphy in psychiatry]. AB - Altered motor activity is a frequently observed symptom in many psychiatric disorders. It is an important diagnostic parameter in mania, depression, ADHD and others. The method which allows measuring motor activity precisely and objectively is actometry. An actometer is a small, electronic device measuring body movements. This paper is a review of psychiatric research in which actometry was used: sleep disorders, ADHD, schizophrenia, anorexia nervosa, affective disorders and also in chronobiology and psychopharmacology. Methodology of actometric research and limitations of the method are discussed. PMID- 15779677 TI - [Course of the EDEN study by using participants' flow sheets]. AB - The development of science allows us to take up new investigative challenges. Interpretations of results can be difficult if there is no adequate and complete information about investigative methods applied, course of trial and statistical analyses. The CONSORT group proposed a standardized diagram facilitating an accurate presentation of the randomised trial. The EDEN study is an example of the application of this method, where the studied groups, number of participants, and all procedures can be presented. It is helpful to prepare reports and find necessary data. The first part of the diagram describes the course of recruitment, the second part presents data necessary to estimate the course of the study and data about numbers of patients in each timepoint whilst the third part shows the assessment of caregivers. CONCLUSIONS: The analysis of the course and correctness of the study is legible if the flow diagram is used. It is useful to find the necessary data for statistical analyses at every stage of study. Permanent control of the study allow to avoid bias upsetting the course of the study and influencing its quality. It introduces comprehensible and straight forward presentation of the results. PMID- 15779678 TI - [59th Annual Meeting of the Society of Biological Psychiatry. April 29-May 1, 2004, New York, New York, USA]. PMID- 15779679 TI - [Cognitive Neuroscience Summer School on Working Memory. Bled, Slowenia 10 17.07.2004]. PMID- 15779681 TI - [Giving life to the years rather than years to life]. PMID- 15779680 TI - Deaths: preliminary data for 2003. AB - OBJECTIVES: This report presents preliminary U.S. data on deaths, death rates, life expectancy, leading causes of death, and infant mortality for the year 2003 by selected characteristics such as age, sex, race, and Hispanic origin. METHODS: Data in this report are based on a large number of deaths comprising approximately 93 percent of the demographic file and 91 percent of the medical file for all deaths in the United States in 2003. The records are weighted to independent control counts for 2003. For certain causes of death such as unintentional injuries, homicides, suicides, and respiratory diseases, preliminary, and final data differ because of the truncated nature of the preliminary file. Comparisons are made with 2002 final data. RESULTS: The age adjusted death rate for the United States decreased from 845.3 deaths per 100,000 population in 2002 to 831.2 deaths per 100,000 population in 2003. Age-adjusted death rates decreased between 2002 and 2003 for the following causes: Diseases of heart, Malignant neoplasms, Cerebrovascular diseases, Accidents (unintentional injuries), Influenza and pneumonia, Intentional self-harm (suicide), Chronic liver disease and cirrhosis, and Pneumonitis due to solids and liquids. They increased between 2002 and 2003 for the following: Alzheimer's disease, Nephritis, nephrotic syndrome and nephrosis, Essential (primary) hypertension and hypertensive renal disease, and Parkinson's disease. Life expectancy at birth rose by 0.3 years to a record high of 77.6 years. PMID- 15779682 TI - [An establishment dedicated to the aged person]. PMID- 15779683 TI - [Complementarity, efficiency and conviviality]. PMID- 15779684 TI - [Evaluating the psychological state and competence in the aged]. PMID- 15779685 TI - [Maintaining a favorable environment in the hospital?]. PMID- 15779688 TI - A practical approach to intersex. AB - The realm of intersex presents a challenge to the family as well the health care provider. As the physical aspects of diagnosis and management have become better understood, it has become apparent that there are psychological and social aspects that we have only begun to understand. A general overview of intersex issues that may confront the urologic practitioner, and the current concepts of diagnosis and management, are provided. PMID- 15779689 TI - Overlapping conditions in women with irritable bowel syndrome. AB - Irritable bowel syndrome is a common and often unpredictable disorder with an increased incidence among women. It is characterized by abdominal pain associated with constipation-diarrhea. Recent research, current theories about etiology and pathophysiology, diagnosis, management, and overlapping conditions such as interstitial cystitis and chronic pelvic pain are discussed. PMID- 15779690 TI - Risk factors for urinary incontinence in African-American women. AB - The majority of research on urinary incontinence (UI) has been conducted with Caucasian populations. This correlational, descriptive study was designed to identify prevalence and risk factors for Ul and to determine the types of incontinence most commonly seen in healthy African-American women. Results of this study showed that age and education were risk factors for Ul in African American women. It is suggested that African-American women be routinely screened for symptoms of Ul as a part of preventive health care. PMID- 15779691 TI - The role of the nurse in the use of vaginal pessaries to treat pelvic organ prolapse and/or urinary incontinence: a literature review. AB - The available literature between 1990 and 2004 was reviewed to determine if the nurse's role in the use of vaginal pessaries to treat pelvic organ prolapse and/or urinary incontinence is well defined. Forty-five articles were reviewed, including one written by a physician's assistant, two written by both a physician and a registered nurse, seven written by registered nurses, 34 written by physicians, and one unpublished manuscript. Nurses could make a valuable contribution to the bank of information available on the use of vaginal pessaries to treat stress urinary incontinence and pelvic organ prolapse. PMID- 15779692 TI - Case and grounded theory as qualitative research methods. AB - Case and grounded theory are two methods of qualitative research. Both methods have their roots in sociology and are focused on understanding, explaining, and/or predicting human behavior. They are ideal methods for nursing research, as they are useful for exploring human responses to health problems. The theoretical underpinnings, methodologies, strategies for data collection, requirements for trustworthiness, and examples of research using case and grounded theory are described. PMID- 15779693 TI - Resolving the nursing shortage: finding passion and joy in nursing. PMID- 15779695 TI - What does it take to become a 'person of influence'? PMID- 15779694 TI - Update on antimicrobial resistance. AB - WHO experts believe that antimicrobial resistance is potentially containable, but the window of opportunity to control and eventually eliminate the most dangerous infectious diseases is closing. If we miss our opportunity, it may become very difficult and expensive--and in some cases impossible--to treat infectious diseases. WHO's global strategy to contain antimicrobial resistance requires a massive effort and an alliance among countries, governments, international organizations, drug manufacturers, and private and public health care sectors. If infectious diseases are fought wisely and widely by the international community, drug resistance can be controlled and lives saved. PMID- 15779696 TI - Getting ready for certification: benign prostatic hyperplasia. PMID- 15779697 TI - Trospium chloride (Sanctura)--new to the U.S. for overactive bladder. PMID- 15779698 TI - Fad diets and obesity--Part IV: Low-carbohydrate vs. low-fat diets. AB - The first three parts of this series of articles covered the basics of some of the more popular low-carbohydrate diets, and the theories behind them. In the fourth and final part of this series, some of the more popular low-fat and low calorie diets, such as the Ornish diet and Weight Watchers, are covered briefly. Recently, several clinical trials of longer duration that compared low carbohydrate versus low-fat diets have been published. These studies demonstrate that some of the low-carbohydrate diets result in reduced weight in the short term, but their ability to reduce weight long-term any better than low-fat or other diets has been questioned. Most popular or fad diets have some positive messages contained within them and some preliminary positive short-term results, but overall the compliance rates with any fad diet are very poor over the long term. The decision to go on any diet should be made with a health professional who can monitor the patient closely. PMID- 15779699 TI - Patient and staff safety. PMID- 15779700 TI - Answering the call for quality nursing care and patient safety: a new model for nursing education. AB - Current discussion on the need to dramatically change nursing education and practice is based on clear concerns about the changing nature of healthcare. The increasingly complex healthcare system of our nation provides sophisticated interventions yet concerns about quality persist. A new model of education and practice is proposed and this model is being implemented through a groundbreaking partnership to prepare a master's educated, entry-level, generalist nursing clinician to lead and guide care at the point of care. The competencies associated with this new clinician are discussed and future implications for evaluation and monitoring are shared. PMID- 15779701 TI - Evaluating the performance and contribution of nurses to achieve an environment of safety. AB - Increasing evidence demonstrates that the quality and stability of nursing personnel substantially influence patient outcomes and healthcare costs. However, standardized performance measures to quantify this influence are limited by the gaps in the understanding of this connection and the underdeveloped state of measure development. Given the importance of nurses and the growing trend to make healthcare quality information available to consumers and purchasers, the paucity of standardized nursing-sensitive performance measures is a major void. To address these needs, the National Quality Forum (NQF) undertook a consensus project to endorse an initial set of performance measures for nursing-sensitive care and to define a research agenda that supports future refreshment and expansion of the national voluntary consensus standards in this area. This article describes the innovative approach taken by NQF to endorse a set of national voluntary consensus standards for nursing-sensitive care and the implications for use. PMID- 15779702 TI - Real-time reporting drives the race toward zero. AB - LifeCare Hospitals of Pittsburgh is a 155-bed freestanding, Acute Long Term Care Hospital, one of 333 nationwide. Patients with complex care needs who are recovering from acute events, postsurgical interventions, and multiple comorbidities stay an average of 25 days. Like other hospitals, reducing medication errors is a focus of the quality improvement program. Provision of a safe environment is tantamount to achieving quality patient outcomes. Creative solutions to the problem of medication errors are needed. Healthcare executives need effective strategies to engage care providers in solving problems in the course of work and in a nonpunitive environment. Over the period of 1 year, medication errors reaching patients at LifeCare decreased by 50% through culture changes, rapid learning cycles, and strong support from the hospital's leaders. PMID- 15779703 TI - Evolution of BCMA within the Department of Veterans Affairs. AB - The Department of Veterans Affairs Medical Center implemented Bar Code Medication Administration (BCMA) between 1999 and 2000 in 161 Medical Centers or Health Systems. BCMA has had a major impact on inpatient licensed nursing staff. Nurses have moved from manual to electronic medication documentation, increasing the complexity of medication administration. There has been acceptance of BCMA by the nurses who are able to see the positive benefits and patient safety aspects. A marked decrease in medication administration errors is a result of implementing BCMA. PMID- 15779704 TI - Nurse's role in tracking adverse drug events: the impact of provider order entry. AB - Adverse drug events (ADE), or injuries caused by drug therapy, are a frequent and serious problem in hospitalized patients. Monitoring, preventing, and treating ADEs is an important patient safety function. Nurses play a significant role in this function, because their data is a unique and important indicator of ADEs and because they are the final point of medication administration. New provider order entry systems with electronic medical records have been viewed as an effective innovation and solution to high rates of ADEs. These systems increase legibility of drug orders, provide decision support, and increase access to the medical record. However, they may not interface with nursing processes effectively. This study reports the experience of a team conducting an ADE surveillance study in a Veterans Health Administration setting where extensive computerized innovations are in place. Lessons learned regarding the integration of nursing work processes with the computerized setting are described. Three areas of concern are highlighted: decreased access to nursing narratives, lack of decision support for medication administration, and failure to code nursing data. Each of these is discussed in terms of relevance to patient safety and the design of information systems. PMID- 15779705 TI - An innovative model for restraint use at the Philadelphia Veterans Affairs Medical Center. AB - In this article, Quality Management Specialists at the Philadelphia Veterans Affairs Medical Center describe an innovative model for restraint use designed to ensure patient safety. They discuss the impetus of the model, its evolution, and purposes. The model provides alternatives to restraint use, education to staff, patients, and families, and electronic tools to monitor restraint usage. Descriptive statistics and analyses of outcomes generated from the electronic tools are presented to demonstrate the utility and value of the model. Future directions for the use of the model and data generated through the electronic record are highlighted. PMID- 15779706 TI - Designing and implementing a close call reporting system. AB - Medical errors and adverse events related to medication errors have received press coverage over the past 3 years. Nursing leaders have by necessity and duty become leaders in the field of patient safety. Defining and reporting errors become critical when analysis of errors relies on adequate and accurate reporting of errors. The next step towards a culture of safety is to avoid "blaming" employees, establishing trust and instituting a close call/near miss reporting system. By encouraging all staff to identify close calls you raise the level of awareness of employees for maintaining a safe patient care environment. Nursing leaders need to guide staff in identifying and reporting close calls through the development and implementation of a transparent reporting system involving recognition and rewarding staff. PMID- 15779707 TI - A business case for patient care ergonomic interventions. AB - This article provides a framework for a business case for patient ergonomic programs that accentuates the financial gains to be realized from such programs as compared to meeting safety requirements. An introduction is made to such commonly used measures as payback period, net present value analysis and internal rate of return. Financial measures on a successful patient handling project in the Veterans Health Administration are outlined and policy implications discussed. PMID- 15779708 TI - Using information to empower nurse managers to become champions for patient safety. AB - There is no longer any question about the risks to patients safety that exist in the hospital. Hospitals are macrosystems that are built upon many interrelated microsystems. Most patient care and hence most errors that directly affect the care outcomes and negatively impact patient safety occur at the microsystem unit level, which is the same level that many improvements to patient safety occur. Patient Safety Net (PSN) is an on-line occurrence reporting tool being used by University HealthSystem Consortium (UHC) member hospitals to report medical events and improve care. As PSN became progressively integrated into the daily operations of these UHC members isolated anecdotes began to surface about how unit nurse managers were able to implement rapid and effective patient safety improvements at the microsystem level on the basis of data received through PSN, without involving performance and safety committees mechanisms. This article highlights the survey performed to validate these improvement anecdotes. PMID- 15779709 TI - Intravenous medication safety system averts high-risk medication errors and provides actionable data. AB - A major responsibility of nursing leaders is to implement systems and policies to improve patient and staff safety, avoid medication errors, and most importantly safeguard patients against harm. In the medication use process, the nurse at the bedside is the most vulnerable, and intravenous (i.v.) drug administration often results in the most serious medication error outcomes. At a 675-bed, tertiary care "Magnet Hospital System," nurses played a key role in a multidisciplinary process that led to successful implementation of a computerized i.v. medication safety system. Software customization, staff training and product set-up were completed in approximately 2 months; 685 devices were installed in 3 hospitals within 12 hours. Nursing acceptance is excellent, and implementation of the system is thought to enhance nursing retention and recruitment. Preliminary data indicate an estimated 849 programming changes ("near misses") annually, ie, potential infusion errors averted by the i.v. medication safety system. A chronogram created from safety data demonstrates that most infusion error warnings occurred between 3:00 PM and 9:00 PM, with an unexpected peak at 6:00 PM. Implementation of the i.v. medication safety system has prevented potentially serious infusion errors and has provided previously unavailable, actionable continuous quality improvement data for best practice improvements. PMID- 15779710 TI - Patient safety: a priority in the US Department of Health and Human Services. AB - This descriptive article provides information about some of the major patient safety initiatives within the operating and staff divisions of the US Department of Health and Human Services. The research for this article was done using the internet. Many health professionals and consumers turn first to the internet while researching a disease or new diagnosis, or while seeking general health information. It is important for nurse administrators to know what resources are readily available to help them implement regulatory requirements, what voluntary programs exist for reporting problems with medical products, what resources are available for consumers to make informed health choices, and where they can get information about specific Department of Health and Human Services programs. PMID- 15779711 TI - Patient and nurse safety: how information technology makes a difference. AB - The Institute of Medicine's landmark report asserted medical error is seldom the fault of individuals, but the result of faulty healthcare policy/procedure systems. Numerous studies have shown that information technology can shore up weak systems. For nursing, information technology plays a key role in protecting patients by eliminating nursing mistakes and protecting nurses by reducing their negative exposure. However, managing information technology is a function of managing the people who use it. This article examines critical issues that impact patient and nurse safety, both physical and professional. It discusses the importance of eliminating the culture of blame, the requirements of process change, how to implement technology in harmony with the organization and the significance of vision. PMID- 15779712 TI - Progress in reducing global measles deaths: 1999-2003. PMID- 15779713 TI - Influenza in the world: September 2004-January 2005. PMID- 15779714 TI - Cigarette tax hike answer to medical assistance deficit? PMID- 15779715 TI - Falls--the great silent killer. PMID- 15779716 TI - Injury: a disease that affects all of society. PMID- 15779717 TI - Public health: a fellowship in government and community service. PMID- 15779718 TI - Making Wisconsin kids SAFE KIDS. PMID- 15779719 TI - Wisconsin's violent death reporting system: monitoring and responding to Wisconsin's violent deaths. AB - In 2003, the Centers for Disease Control and Prevention (CDC) awarded the Wisconsin Department of Health and Family Services, Injury Prevention Program, a grant to participate in a multistate project called the National Violent Death Reporting System (NVDRS). The purpose of the Wisconsin Violent Death Reporting System (WVDRS) is to link violent death records (death certificates, police reports, medical examiner and coroner reports, crime laboratories, and perhaps child fatality review teams) from the same event, promote more timely information retrieval, describe in detail circumstances that may have contributed to the violent death, and identify and characterize perpetrators and their relationships to victims. This article describes the development of WVDRS and its importance in understanding and preventing violent injury and death in Wisconsin. PMID- 15779720 TI - Trends, risk factors, and prevention of falls in older adults in Wisconsin. AB - Wisconsin's death rate due to falls among adults 65 years and older is more than twice the national average. The hospitalization rate due to falls-related injuries in Wisconsin increased slightly from 1995 to 2002, with an injury rate of 2159 per 100,000 in 1995, and 2263 per 100,000 in 2002. Emergency department (ED) utilization and hospitalization rates for falls-related injury are higher for women than for men in Wisconsin. In 2002, the total statewide charges for hospitalizations and ED visits for falls-related injuries were more than $96 million. Two thirds of those admitted to the hospital for a falls-related injury were discharged to a nursing home or rehabilitation facility. Multifactorial intervention strategies have been shown to decrease the rate of falls in randomized, controlled trials. The purpose of this paper is to describe trends in falls-related injury fatalities, hospitalizations, and ED visits in Wisconsin. Also included are cost data related to falls, identification of risk factors, and descriptions of the possible role of physicians and other health care professionals in interdisciplinary, multifocal programs to prevent falls-related injuries in high-risk older adults. PMID- 15779721 TI - Community-based fall prevention programs for older adults in Wisconsin. AB - Falls prevention and intervention programs are important strategies to reduce the considerable morbidity and mortality from falls in older adults in Wisconsin. This article attempts to dispel the common misperception that falls are a normal part of the aging process, to provide examples of prevention programs in Wisconsin targeted specifically to community-dwelling older adults, and to provide prevention messages for older adults and the health care and social or human services professionals that serve them. PMID- 15779722 TI - Kids can't fly: preventing fall injuries in children. AB - Falls are the leading cause of unintentional injury in children. Pediatric fall injuries can be grouped into roughly 4 categories: (1) falls while walking or running, (2) falls from heights, (3) falls while participating in recreational activities, and (4) falls during competitive sports activities. Because injury statistics collected by the US Consumer Product Safety Commission are product related, there is little data on falls from walking or running. Falling down stairs is the most common height-related fall injury, followed by playground related falls. Bicycle-related injury leads the third category in frequency. Over 30% of parents report that their child has been injured while playing a team sport; although not all sports-related injuries are caused by falling, most result in a fall. Strategies for preventing fall injuries are based on finding risk factors that can be modified; these modifications can be pre-event, event specific, or post-event. Office-based injury prevention counseling plays an important role in preventing pediatric fall injury. PMID- 15779723 TI - Give Wisconsin children a boost. AB - In the United States, more children die from motor vehicle crashes than any other cause. Research has demonstrated that children ages 4-8 have a significantly reduced risk of injury if they are restrained in booster seats rather than adult seatbelts. Despite current recommendations, few children in this age group are properly restrained. Health care providers can help increase booster seat use by educating parents, participating in community campaigns, and advocating for mandatory booster seat laws. PMID- 15779724 TI - Analysis of booster seat and seat belt use: how many Wisconsin childhood deaths and hospitalizations could have been prevented in 1998--2002? AB - BACKGROUND: Motor vehicle collisions (MVC) remain the leading cause of childhood death and injury in Wisconsin and throughout the nation. The objective of this study is to estimate the potential benefits of increased use of belt-positioning booster (BPB) seats for children ages 4-7, and increased seat belt use for children ages 8-18.5 years. The outcomes measured were the reduction in MVC associated childhood deaths and hospitalizations as a result of increased use of BPB seats or seat belts. METHOD: Childhood MVC-associated deaths and hospitalizations from 1998 to 2002 were obtained from the Wisconsin Interactive Statistics on Health (WISH) data query system. Three separate age groups were analyzed: children ages 4-7, 8-15, and 16-18.5 years. Ages for the groups were designed to match current "best practice" recommendations for the use of BPB seats, published age-range specific baseline restraint use data, and age specific mortality/injury relative risk (RR) values as closely as possible for a pediatric population. Population attributable risk (PAR) was calculated to predict preventable deaths and injuries (hospitalizations) over the years studied. RESULTS: From 1998 to 2002, MVCs accounted for 440 childhood deaths and 2639 injuries requiring hospital admission. Using the calculated PAR population statistic for each age group studied, the predicted reduction in childhood MVC deaths and hospitalizations were calculated for various increases in restraint use above current baseline compliance. At the 100% use level, the model predicted 16 fewer deaths and 84 fewer hospitalizations for children 4-7 years old (BPB seat use); 45 fewer deaths and 206 fewer hospitalizations for children 8-15 years old (seat belt use); and 119 fewer deaths and 669 fewer hospitalizations for children 16-18.5 years old (seat belt use). The total potentially avoided deaths and hospitalizations across all age groups studied was 180 childhood deaths and 959 hospitalizations during this 5-year study period. CONCLUSION: Communities should devote resources to public education programs directed at increasing the use of belt-positioning booster seats for children 4-7 years old and seat belts for older children. Evidence-based child passenger safety laws should be improved and enforced in Wisconsin in an effort to support families in their attempts to safeguard their children. With a multi-component child passenger restraint safety campaign, unnecessary MVC-associated childhood deaths and injuries can be avoided in the future. PMID- 15779725 TI - Improving awareness and use of booster seats in Head Start families. AB - OBJECTIVES: To determine the knowledge level of Head Start providers, parents, and students about booster seats and to directly observe booster seat use before and after a combined educational program and booster seat giveaway. METHODS: Before and after a short educational session and child safety seat giveaway, Head Start providers and parents received a brief questionnaire on booster seats and the state child restraint law. Direct parking-lot observation of booster seat use was performed before and after the giveaway. RESULTS: Forty-three students were enrolled in the study, with 33 receiving booster seats and 5 receiving forward facing car seats, dependent on the weight and age of the child. Before the study, 15 (35%) of the children had weight/age appropriate child safety seats; after the giveaway, this number increased to 42 (98%; P<0.001). The proportion of children observed using booster seats before the giveaway was 6%, which increased to 34% after the giveaway (P<0.001). CONCLUSIONS: This study indicates that a booster seat giveaway can be successful in increasing the number of children who use booster seats; however, the majority (66%) of participating children still rode inappropriately restrained after the giveaway. Steps beyond providing booster seats, such as combining this intervention with ongoing parent and community education efforts, as well as legislation and enforcement, are needed to bring booster seat use to a high level. The information from this study may be helpful in designing future programs intended to increase booster seat use, as well as emphasizing the need for booster seat legislation. PMID- 15779726 TI - Wisconsin's experience with the Graduated Driver Licensing Law. AB - In 2000, Wisconsin enacted the Graduated Driver Licensing (GDL) Law in an effort to reduce the number of teen deaths and injuries due to motor vehicle crashes. The GDL program targets high-risk teen driving behaviors by mandating more hours of supervised driving, limiting nighttime driving, restricting vehicle passengers, requiring seatbelt use, and mandating absolute sobriety for all teen drivers. In the first 3 years of graduated driver licensing, Wisconsin saw a 15% decrease in the number of motor vehicle collisions involving 16-year-old drivers. While this decline is encouraging, it lags behind the effectiveness documented by other states with GDL laws. The current study evaluates informational or attitudinal barriers that may prevent the effective implementation of Wisconsin's GDL law. Knowledge and attitude surveys were administered to 15-17 year olds and their parents. Parents (n=26) and teens (n=26) were knowledgeable about the 30 hours of supervised driving during the learner's permit phase and extremely knowledgeable about the 9-month nighttime and passenger restrictions of the probationary phase. While 76% of parents supported GDL, 70% of teens disapproved of the law. The reasons for disapproval varied, parents often cited inconvenience and restrictiveness. Teens most often cited passenger restrictions as the reason for disapproval. Overall, negative attitudes, more than lack of knowledge, may weaken compliance with Wisconsin's GDL law and, in turn, limit the reduction of teen crashes. PMID- 15779727 TI - Bullying prevention: Wisconsin takes a stand. PMID- 15779728 TI - The burden of suicide and homicide of Wisconsin's children and youth. AB - OBJECTIVE: To describe the epidemiology of violent deaths (suicides and homicides) of children and youth in Wisconsin. METHODS: Linked data from death certificates, coroners or medical examiners, Uniform Crime Reports, police case reports and crime laboratories were analyzed using SPSS 11.5. Analyses from 2000 to 2002 describe suicides and homicides of children and youth under age 25. Rates are per 100,000 population per year. RESULTS: A total of 585 persons under age 25 in Wisconsin died from suicide or homicide during 2000--2002. Suicides outnumbered homicides 323 (55%) to 262 (45%). Firearms were involved in 59% (n=344) of cases. Fatality rates increased with advancing age, ranging from 2.0 in 0-13 year olds to 26.9 in youth age 21-24. There were over twice as many violent deaths compared to cancer and infectious disease deaths combined (n=253). CONCLUSION: Suicides and homicides are a significant burden on the children and youth of Wisconsin. An investment in reducing this burden requires comprehensive data and informed programs and policies. We recommend that physicians and public health community leaders collaborate with the criminal justice community and policy makers to develop, implement, and evaluate prevention programs and policies. PMID- 15779730 TI - What is managed money and should you care? PMID- 15779729 TI - Medical College faculty provide national leadership in injury control and prevention. PMID- 15779731 TI - The surgical care improvement project. PMID- 15779732 TI - Osteosarcoma of the humerus with removal of the arm, clavicle and scapula; death from sarcoma of lung nine months later. 1904. PMID- 15779733 TI - Medical-surgical nursing: a specialty or not? PMID- 15779734 TI - A quest for quality: from data to care delivery. PMID- 15779735 TI - The influence of race on heart failure in African-American women. AB - Numerous factors contribute to the rate of heart failure in African-American women. More research is needed to elucidate and address heart failure in this population. PMID- 15779736 TI - Assessment and treatment of terminal restlessness in the hospitalized adult patient with cancer. AB - Terminal restlessness affects a large proportion of patients with cancer at the end of life. It has many different risk factors, presentations, and causes. Management consists of keen assessment, identification and reversal of the causes, and treatment by a combination of pharmacological, environmental, and spiritual interventions. PMID- 15779737 TI - A nursing standards-based system that works! AB - A 200-bed community hospital has been using a standards-based system since 1985 to guide nursing practice and establish the framework for documentation. Managers and staff nurses appreciate this system for its comprehensive yet simple approach, and its value has stood the test of time. The comprehensive standards model and its use are described. PMID- 15779738 TI - Immobility: geriatric self-learning module. AB - The Geriatric Resource Nurse Model is used at the University of Virginia to improve the competency of staff in caring for older adults. Eight self-learning educational modules were developed to address common concerns in hospitalized elders. The Immobility. Geriatric Self-Learning Module is published here, along with a post-test. This is the third in a four-part publication of self-learning modules. PMID- 15779739 TI - Confusion: geriatric self-learning module. AB - The Geriatric Resource Nurse Model is used at the University of Virginia to improve the competency of staff in caring for older adults. Eight self-learning educational modules were developed to address common concerns in hospitalized elders. The Confusion. Geriatric Self-Learning Module is published here, along with a post-test. This is the third in a four-part publication of self-learning modules. PMID- 15779740 TI - Update on risk factors for atherosclerosis: the role of inflammation and apolipoprotein E. AB - Over 1 million adults will have a new or recurrent myocardial infarction this year. Traditional risk factor assessment predicts less than one-half of all future cardiovascular events, and many patients develop atherosclerosis in the absence of these factors. Alternative risk factors, including genotype and the inflammatory response, are presented, along with intervention considerations for the medical-surgical nurse. PMID- 15779741 TI - The lived experience of having a chronic wound: a phenomenologic study. AB - Though millions of American elders suffer from chronic non-healing wounds, relatively little research has been done on the wound experience. The purpose of this descriptive phenomenologic study was to describe the lived experience of having a chronic wound. PMID- 15779742 TI - What contributes to professionalism? AB - Appearance, behavior, dress, and communication skills play an important role in the image that a nurse projects. As the nurse interacts with patients, families, community members, corporate personnel, and policymakers, he or she must reflect a professional image. PMID- 15779743 TI - Management of gastrointestinal fistulas: a case study. AB - The management of GI fistulas can be very challenging for the health care team and the patient. The treatment goals are to provide bowel rest, prevent fluid and electrolyte imbalances, protect perifistula skin, and contain the effluent. Consistent nursing care and team collaboration can lead to successful patient outcomes. PMID- 15779744 TI - Enlisting medical-surgical nurses in the battle against cancer. PMID- 15779746 TI - Prediction of large variation in biota to sediment accumulation factors due to concentration-dependent black carbon adsorption of planar hydrophobic organic compounds. AB - Biota-to-sediment accumulation factors (BSAFs) sometimes show large variations between sites for a given planar hydrophobic organic compound (pHOC) in a given organism. We present a framework in which strong adsorption to soot/charcoal (black carbon [BC]) explains such BSAF variations. Recently constrained parameters on environmental BC sorption were used in a dual-mode sorption model of BSAE Variations in sedimentary BC content are shown to explain variations in BSAF of one to two orders of magnitude. In addition, strong BC sorption can explain the often-observed difference in BSAF between polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs). The strong nonlinearity of BC sorption can render BSAF values strongly concentration-dependent for a given sediment/organism/compound combination, so it is of paramount importance to carry out uptake and toxicity tests at the relevant environmental concentrations. PMID- 15779747 TI - Soil-solution speciation of Cd as affected by soil characteristics in unpolluted and polluted soils. AB - Total metal content by itself is insufficient as a measure to indicate actual environmental risk. Understanding the mobility of heavy metals in the soil and their speciation in the soil solution is of great importance for accurately assessing environmental risks posed by these metals. In a first explorative study, the effects of general soil characteristics on Cd mobility were evaluated and expressed in the form of empirical formulations. The most important factors influencing mobility of Cd proved to be pH and total soil content. This may indicate that current legislation expressing the requirement for soil sanitation in Flanders (Belgium) as a function of total soil content, organic matter, and clay does not successfully reflect actual risks. Current legal frameworks focusing on total content, therefore, should be amended with criteria that are indicative of metal mobility and availability and are based on physicochemical soil properties. In addition, soil-solution speciation was performed using two independent software packages (Visual Minteq 2.23 and Windermere Humic Aqueous model VI [WHAM VI]). Both programs largely were in agreement in concern to Cd speciation in all 29 soils under study. Depending on soil type, free ion and the organically complexed forms were the most abundant species. Additional inorganic soluble species were sulfates and chlorides. Minor species in solution were in the form of nitrates, hydroxides, and carbonates, the relative importance of which was deemed insignificant in comparison to the four major species. PMID- 15779748 TI - Aging of triazine amine in soils demonstrated through sorption, desorption, and bioavailability measurements. AB - The aging of triazine amine in soil was studied during a time course of 119 d by measuring bioavailability in terms of mineralization after inoculation of the triazine amine-degrading bacterium Rhodococcus erythropolis TA57. The bioavailability was measured in four soil samples: A-, B-, and C-horizons from an agricultural soil profile and in a peat soil. The sorption of triazine amine in the soil samples was quantified during the period of aging in terms of sorption distribution coefficients (Kd) and desorption distribution coefficients (Kd,des). Measures of bioavailability and triazine amine concentration in the nonavailable fraction showed effects of aging in the soils that were rich in organic matter. The triazine amine bioavailability declined significantly during the aging period in soils containing greater than 2% organic carbon, whereas the B- and C-horizons showed no signs of aging, in agreement with their low content of organic material. Corresponding to this, desorption decreased significantly in the A horizon but, surprisingly, not in the peat soil. Analyses by thin-layer chromatography indicated an association of aqueous triazine amine and dissolved organic matter in the peat soil. This gives an explanation for both the significant decrease in bioavailability and the noncorresponding stability of the nonavailable (i.e., nondesorbed) fraction. PMID- 15779749 TI - Aqueous photochemistry of triclosan: formation of 2,4-dichlorophenol, 2,8 dichlorodibenzo-p-dioxin, and oligomerization products. AB - The photochemical fate of the antimicrobial agent triclosan is presented. Experiments performed in both natural and buffered deionized water show that triclosan rapidly photodegrades by direct photolysis (t(1/2) = 5 h, pH 8, noon summer sunlight, 45 degrees N latitude). Both 2,8-dichlorodibenzo-p-dioxin (2,8 DCDD) and 2,4-dichlorophenol (2,4-DCP) are produced. The 2,8-DCDD and 2,4-DCP also are photolabile and, thus, are intermediates. The yields for 2,8-DCDD and 2,4-DCP ranged from 3 to 12% depending on the conditions employed. When triclosan is photolyzed in the presence of Suwannee River (GA, USA) fulvic acid, a portion of the initial mass is recovered as insoluble material. Based on experiments in which the formation of insoluble material was monitored with photolysis time, it is postulated that photolysis in natural waters leads to some of the triclosan being coupled to humic matter. Triclosan also reacts rapidly with both singlet oxygen (k(rxn) = 1.07 +/- 0.03 x 10(8) M(-1) s(-1) in water of pH 10) and hydroxyl radical (k(*OH) = 5.4 +/- 0.3 X 10(9) M(-1)(s-1). Indirect photolysis pathways, however, are not expected to be important because of low steady-state concentrations of reactive oxygen species in natural waters and the efficiency of the direct photolysis of triclosan. PMID- 15779750 TI - Fate of mercury accumulated by blowflies feeding on fish carcasses. AB - Because fish represent the principal methylmercury (MeHg) pool in the water column of freshwater systems, MeHg released from their carcasses could represent an important flux to the environment. Necrophagous invertebrates such as fly larvae can play an important role in this Hg recycling. We studied Hg accumulation by blowflies (Diptera, Calliphoridae) feeding on beached fish carcasses in the field. We found that the MeHg these flies accumulated as larvae is retained in their pupal stage but is eliminated by the adult following emergence. We conclude that calliphorids possess an efficient mechanism for excreting the MeHg that they accumulate from carcasses. PMID- 15779751 TI - Comparison of different microbial bioassays to assess metal-contaminated soils. AB - These experiments compared the sensitivity of four different types of bioassay over time after five metals were added to a wide range of soils at the maximum concentrations in the European Union Sewage Sludge Directive. Three were chronic assays (most probable number of Rhizobium leguminosarum, soil microbial C and Biolog substrate utilization). The fourth bioassay, an acute biosensor, employed a lux-marked luminescent bacterium (Escherichia coli) in the soil pore water. Five metals were added to 23 different soils as a mixture at Zn = 300, Cd = 3, Pb = 300, Cu = 135, and Ni = 75 mg/kg as nitrate salts and compared with unamended controls. Zinc and Cu were the metals most likely to be toxic at the concentrations used here. In the case of Rhizobium, the number of cells in soil was not affected after 11 d; however, by 818 d the numbers had decreased by four orders of magnitude with increasing concentrations of Zn and Cu in soil solution. Microbial biomass also was not affected after 11 d, but significantly decreased with increased Zn (p < 0.001) and Cu (p < 0.01) in soil solution after 818 d. Toxicity to the soil microbial biomass increased with time, whereas the toxicity to the biosensor remained the same. Biolog substrate utilization profiles were not responsive to the concentrations used here. PMID- 15779752 TI - Effect of interaction between density dependence and toxicant exposure on population growth rate of the potworm Enchytraeus doerjesi. AB - Laboratory cultures of the potworm Enchytreaus doerjesi were exposed to zinc contaminated soil at nominal concentrations 0, 75, 150, 300, or 600 mg Zn/kg dry weight for four weeks (one generation) at five different initial densities (5, 10, 20, 40, or 80 individuals per 20 g soil). The final culture numbers and the instantaneous population growth rate, r(i) were used as measures of culture response to both factors. In all treatments, the number of individuals increased during the experiment; thus, all the cultures exhibited positive r(i) values. The population growth rates decreased significantly with increasing Zn concentration and were also affected by the initial density. However, the latter effect was nonlinear and the initial density of 20 individuals resulted in the highest r(i) values throughout the whole range of Zn concentrations tested, while the lowest population growth rates were recorded for the highest initial density. Significant interaction between zinc toxicity and population size was found for the two lowest initial densities, where synergistic effects were noted. No effect of density on zinc toxicity was found at initial densities of 20 to 80 individuals, most probably because the animals were fed ad libitum and thus were not food limited. PMID- 15779753 TI - Field validation of sediment zinc toxicity. AB - A field study was conducted to validate concentrations of zinc in freshwater sediments that are tolerated by benthic macroinvertebrate communities and to determine whether a relationship exists with the acid volatile sulfide (AVS) simultaneously extracted metal (SEM) model. In both the lake and riverine systems, one sediment type was high in AVS and one low in AVS, which resulted in zinc-spiked sediments that ranged from low to high SEM to AVS ratios. The colonization trays were sampled seasonally, ranging from 6 to 37 weeks of exposure, and were evaluated using several appropriate benthic indices. Results of the field evaluations at the four test sites confirmed the validity of the AVS SEM model, predicting benthic macroinvertebrate effects correctly 92% of the time. In sediments where the SEM to AVS ratio or the AVS and organic (OC) normalized fractions exceeded 8 and 583 micromol/g of OC, toxicity was observed from the zinc-spiked sediments. Conversely, when the SEM to AVS ratio or OC normalized AVS fractions were less than 2 or 100 micromol/g of OC, no toxicity was observed. In the range of 148 to 154 micromol/g of OC, toxicity varied in two treatments. Total zinc concentrations in sediments showed no relationship to benthic effects. The most impaired benthic community occurred in the high gradient stream sediments, which had low OC and AVS concentrations and SEM to AVS ratios of 33 and 44 in the spiked sediments. Five to six benthic metrics were depressed at SEM to AVS ratios of 8.32 and 9.73. The no-observed-effect level appeared to be near a SEM to AVS ratio of 2, with slight to no effects between ratios of 2.34 and 2.94. No sites with ratios of less than 2 showed any adverse effects. PMID- 15779754 TI - Bioaccumulation of metals in Eisenia fetida after exposure to a metal-loaded bauxsol reagent. AB - The present study investigated the acute toxic effects of a metal-loaded Bauxsol reagent (MLBR), containing more than 6,450 mg kg(-1) of bound metal, on the earthworm Eisenia fetida. Bauxsol is manufactured by Virotec Global Solutions of Gold Coast, Queensland, Australia. The E. fetida were exposed to 0, 10, 20, 40, 60, and 80% treatments of MLBR plus cow manure for 28 d. Good motility and no mortality of E. fetida were found in all treatments after 28 d of exposure. The greatest bioaccumulation of metals by E. fetida occurred in the 20 and 40% MLBR treatments. The bioaccumulation of metals in the worms and bioaccumulation factors (BAFs) were less than the reported toxicity thresholds to cause mortality and less than the reported BAF for moderately contaminated soils, indicating that metals bound to Bauxsol reagents are mostly nonbioavailable. Analyses of the 20% MLBR treatment at 28 d using a sequential extraction procedure showed that more than 95% of the metals are bound within the Fe/Mn oxide fractions. However, changes occurred in metal fractionation after exposure to E. fetida for Cd/Cr, and Fe/Mn. The data also showed that the exchangeable (1 M MgCl2) and the toxicity characteristic leaching procedure extractant are useful as indicators of metal bioavailability from MLBR to E. fetida. PMID- 15779755 TI - Projected population-level effects of thiobencarb exposure on the mysid, Americamysis bahia, and extinction probability in a concentration-decay exposure system. AB - Population-level effects of the mysid, Americamysis bahia, exposed to varying thiobencarb concentrations were estimated using stage-structured matrix models. A deterministic density-independent matrix model estimated the decrease in population growth rate (lambda) with increasing thiobencarb concentration. An elasticity analysis determined that survival of middle stages provided the largest contribution to lambda. Decomposing the effects of lambda in terms of changes in the matrix components determined that reduced reproduction had a large influence on population dynamics at lower thiobencarb concentrations, whereas reduced survivorship had the largest impact on populations at higher concentrations. A simulation model of a concentration-decay system was developed to demonstrate the importance of integrating chemical half-life and management practices in determining population viability. In this model, mysids were originally exposed to a high thiobencarb concentration (300 microg/L) that decayed an order of magnitude in the number of mysid generations corresponding to thiobencarb half-life values under three different exposure regimes. Environmental stochasticity was added to the model to estimate the cumulative extinction probability of mysids exposed to fluctuating concentrations of thiobencarb in random environments. The cumulative extinction probability increased with thiobencarb half-life, stochasticity, and concentration present at the time of a new exposure. The model demonstrated the expansion of population projection models in determining the ecological impact of a population exposed to pesticides. PMID- 15779757 TI - Macroinvertebrate community response to pulse exposure with the insecticide lambda-cyhalothrin using in-stream mesocosms. AB - Pesticides are constantly being applied to agricultural catchments, but little is known about their impact on aquatic biota during natural exposure. In the present study, the impact of the pyrethroid lambda-cyhalothrin was studied in an in stream mesocosm setup. Twice during the summer of 2002, the natural macroinvertebrate community was exposed in situ to a 30-min pulse of lambda cyhalothrin. Pyrethroid doses were released through a modified drip set with nominal concentrations of 0.10, 1.00, and 10.0 microg L(-1) during the first exposure and 0.05, 0.50, and 5.00 microg L(-1) in the second exposure. Before, during, and after exposure, drifting macroinvertebrates were caught in nets. Quantitative benthic samples were taken both before and on two occasions after exposure. Macroinvertebrate drift increased immediately after the pulse exposure, with total drift being significantly higher at all concentrations. Gammarus pulex, various Ephemeroptera, Leuctra sp., and Simuliidae were some of the taxa showing the most pronounced drift response. Structural change in the community was found only at 5.00 and 10.0 microg L(-1), and recovery occurred within approximately two weeks. The present study may be valuable in assessing extrapolations based on laboratory results as well as in evaluating pyrethroid impact on natural freshwater environments. PMID- 15779756 TI - Biofilm formation by algae as a mechanism for surviving on mine tailings. AB - Photosynthetic biofilms successfully colonize the sediments of a mine tailings reservoir (Guanajuato, Mexico) despite the high metal concentrations that are present. To elucidate the mechanisms of biofilm survival despite metal ores, experiments were performed to evaluate the response of seminatural biofilms to Cu, Zn, and a combination of both metals at concentrations observed in the field. The biofilms were composed mostly of the chlorophyte Chlorococcum sp. and the cyanobacterium Phormidium sp., and their response to the two added metals was described by measurements of extracellular polymeric substances (EPS) and in vivo fluorescence. The photosynthetic efficiency and the minimal chlorophyll fluorescence of dark-adapted cells were measured by multiwavelength pulse amplitude-modulated fluorometry. The photosynthetic efficiency of light-adapted cells (phi(PSII)) also was measured. Metal exposure increased the EPS production of biofilms, as visualized with confocal laser-scanning microscopy. Extracellular polymeric substances enhanced the extracellular metal accumulation from the first day of metal exposure. Metals provoked changes in the relative abundance of the dominant taxa because of a species-specific response to the metals when added individually. Metals affected the phi(PSII) less than the total biomass, suggesting ongoing activity of the surviving biofilms. Survival of individual biofilm photosynthetic cells was found to resume from the embedding in the mucilaginous structure, which immobilizes the metals extracellularly. The survival of biofilms under mixed-metal exposure has practical applications in the remediation of mine tailings. PMID- 15779758 TI - Early-life-stage toxicity in offspring from exposed parent medaka, Oryzias latipes, to mixtures of tributyltin and polychlorinated biphenyls. AB - The present study examined the effects of tributyltin (TBT), polychlorinated biphenyls (PCBs), and mixtures of both chemicals on reproduction in Japanese medaka, Oryzias latipes. For 21 d we gave groups of medaka freeze-dried brine shrimp flakes contaminated with a mixture of either 0, 1, 5, or 25 microg TBT g( 1) plus 0 or 25 .micro PCBs g(-1). We measured the fecundity and fertility of the parent fish and assessed the deformity, hatchability, time-to-hatching, and swim up failure rate of the next generation. Fertilization success in the third week of the administration period was significantly decreased by administration of 25 microg TBT g(-1) (77%) compared with the control group (87%). Both TBT and PCBs were transferred maternally into the eggs of the next generation, causing early life-stage toxicity. Administration of 1 microg TBT g (-1) was not toxic to embryological development, but abnormal eye development (i.e., small eyes or no eyes) occurred when TBT at the same concentration was mixed with PCBs (6.4%). Administration of TBT alone significantly decreased hatchability and increased swim-up failure, and administration of PCBs alone significantly increased time-to hatching. Statistical analysis by two-way analysis of variance detected an interaction between TBT and PCBs in these three parameters. TBT induces abnormal development of the eyes, reduced hatchability, and increased swim-up failure, whereas PCBs delay time-to-hatching. Administration of mixtures of TBT and PCBs has more adverse effects on the developmental stage of medaka than does that of each chemical alone. PMID- 15779759 TI - Uptake and tissue-specific distribution of selected polychlorinated biphenyls in developing chicken embryos. AB - Fertilized chicken eggs were injected with high doses of individual polychlorinated biphenyl (PCB) congeners (0.5 microg of PCB 77, 9.8 microg of PCB 153, or 10.9 microg of PCB 180) before incubation to investigate the structure specific uptake of these compounds by the embryo and their accumulation in brain and liver tissue. In accordance with earlier publications, a gradual uptake and accumulation of these compounds was observed during the last week of embryonic development. The PCB uptake and distribution to the specific tissues did not appear to be structure dependent. Wet-weight liver PCB concentrations (18, 266, and 278 ng/g at hatching for PCB 77, PCB 153, and PCB 180, respectively) were consistently two- to fourfold higher than carcass levels (7 ng/g of PCB 77, 117 ng/g of PCB 153, and 81 ng/g of PCB 180 at hatching). Whereas liver and carcass concentrations increased exponentially between day 13 of incubation and hatching, PCB levels in brain tissue remained unaltered (range, 0.6-1.0 ng/g of PCB 77 and 8-12 ng/g of PCB 153 and PCB 180 throughout the last week of incubation). Lipid analysis of the organs suggested that the lipid composition of brain may be an important factor explaining the low PCB accumulation in this tissue. PMID- 15779760 TI - Influence of dimethoate on acetylcholinesterase activity and locomotor function in terrestrial isopods. AB - Locomotor behavior in terrestrial organisms is crucial for burrowing, avoiding predators, food seeking, migration, and reproduction; therefore, it is a parameter with ecological relevance. Acetylcholinesterase (AChE) is a nervous system enzyme inhibited by several compounds and widely used as an exposure biomarker in several organisms. Moreover, changes in energy reserves also may indicate an exposure to a stress situation. The aim of this study is to link biomarkers of different levels of biological organization in isopods exposed to increasing doses of dimethoate in semifield conditions. Locomotor parameters, AChE activity, and energy reserves (lipid, glycogen, and protein contents) were evaluated in the isopod Porcellio dilatatus after 48-h and 10-d exposure to dimethoate-contaminated soil. Results showed a clear impairment of both locomotor and AChE activity during the entire study, although effects were more pronounced after 48 h. Most locomotor parameters and AChE activity showed a clear dose response relationship. By contrast, no clear trend was observed on energetic components. A positive and significant relationship was found between AChE activity and those locomotor parameters indicating activity, and the opposite was observed with those locomotor parameters indicating confusion and disorientation. The results obtained in this study enhance the importance of linking biochemical responses to parameters with ecological relevance at individual level, the value of locomotor behavior as an important marker to assess effects of toxicants, and also the usefulness and the acquisition of ecological relevance by AChE as a biomarker, by linking it with ecologically relevant behavioral parameters. PMID- 15779761 TI - Thyroid hormone status in gray seal (Halichoerus grypus) pups from the Baltic Sea and the Atlantic Ocean in relation to organochlorine pollutants. AB - Marine apex predators, such as the gray seal (Halichoerus grypus), are exposed to high concentrations of organochlorine (OC) compounds via biomagnification in the food web. These lipophilic pollutants are transferred from the female to her offspring during lactation, and there is particular concern for endocrine disruption in the offspring. Herein we examined associations between the blubber concentrations of polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and metabolites, chlordanes, hexachlorocyclohexanes (HCHs) and hexachlorobenzene (HCB), and plasma concentrations of thyroid hormones (thyroxine [T4] and tri-iodothyronine [T3]) in free-ranging, newly weaned gray seal pups from the Baltic Sea and the Norwegian waters of the Atlantic Ocean. Concentrations of total T3 (TT3) and free T3 (FT3) were significantly lower in the polluted seals from the Baltic Sea. Blubber concentrations of the sum of the analyzed PCB congeners (sigmaPCBs) and the sum of the DDT compounds (sigmaDDTs) were correlated negatively with plasma TT3 and FT3 concentrations. The results from this study indicate that OCs may affect levels of thyroid hormones in free-ranging gray seal pups, and that more focus should be put on T3 when addressing the possible effects of OCs on thyroid hormone function and status in nonrodent species. PMID- 15779762 TI - Effects of contaminant exposure on reproductive success of ospreys (Pandion haliaetus) nesting in Delaware River and Bay, USA. AB - Despite serious water-quality problems and pollutant loading and retention, Delaware River and Bay (USA) provide important wildlife habitat. In 2002, we conducted a comprehensive evaluation of contaminant exposure and reproduction of ospreys (Pandion haliaetus) breeding in Delaware River and Bay. Sample eggs were collected from 39 nests and analyzed for organochlorine pesticides, polychlorinated biphenyls (PCBs), and mercury; a subset of 15 eggs was analyzed for perfluorinated compounds and polybrominated diphenyl ethers (PBDEs). The fate of each nest was monitored weekly. Concentrations of 10 organochlorine pesticides or metabolites, total PCBs, and several toxic PCB congeners were greater (p < 0.05) in eggs collected between the Chesapeake and Delaware Canal (C and D Canal) and Trenton (Delaware River and northern Bay) compared to other sites. Concentrations of p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE; 0.785-3.84 microg/g wet wt) and total PCBs (5.50-14.5 microg/g wet wt) in eggs collected between the C and D Canal and Trenton were similar to levels recently found in the Chesapeake Bay. In all study segments, at least one young fledged from 66 to 75% of nests. Productivity for Delaware Inland Bays (reference area) and southern Delaware Bay was 1.17 and 1.42 fledglings/active nest, respectively; north of the C and D Canal, productivity was 1.00 fledgling/active nest, which is marginally adequate to maintain the population. Using these data, a logistic regression model found that contaminant concentrations (p,p'-DDE, heptachlor epoxide, chlordane and metabolites, and total PCBs) were predictive of hatching success. Several perfluorinated compounds and PBDEs were detected in eggs at concentrations approaching 1 microg/g wet weight. These findings provide evidence that contaminants continue to be a significant stressor on osprey productivity in the northern Delaware River and Bay. PMID- 15779763 TI - Polychlorinated biphenyls, dioxins, and furans in weaned, free-ranging northern elephant seal pups from central California, USA. AB - The aim of this study was to measure persistent organic pollutants in northern elephant seals ([NES], Mirounga angustirotris). We obtained blubber biopsy samples from six healthy, newly weaned NES pups from Ano Nuevo, California (USA). Contaminant levels were lower than those of other pinnipeds studied on the west coast of North America. Blubber concentrations of polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans averaged 700 +/- 130 microg/kg, 32 +/- 23 ng/kg, and 17 +/- 5 ng/kg (lipid wt), respectively. These contaminants originate from transplacental transfer and from maternal milk, which, in turn, reflect contaminants acquired by the mother from prey during long-distance foraging trips in the northeastern Pacific Ocean. The PCB profile in the blubber of NES pups mainly was composed of penta-, hexa-, and hepta-chlorobiphenyls, possibly reflecting the deep-sea nature of the mother's diet. Our results suggest that NES pups, in contrast to pups of other pinnipeds in the eastern Pacific Ocean, are exposed to low levels of persistent organic pollutants, reflecting an open ocean signal. PMID- 15779764 TI - Toxicity of heavy metals in soil assessed with various soil microbial and plant growth assays: a comparative study. AB - Abstract-Elevated metal concentrations in soils can disturb the soil ecosystem; thus, researchers strive to identify the most sensitive assay for detection of the early signs of toxicity. The purpose of the present study was to compare eight different ecotoxicological endpoints on the same set of metal-contaminated soils that were collected from seven series of soils sampled during field trials. The endpoints are based on three microbial assays (potential nitrification rate [PNR], substrate-induced respiration [SIR], and basal respiration [BR]) and two plant growth tests, one of which included symbiotic N fixation. The overall sensitivity of the endpoints to detect statistically significant adverse effects ranked as follows: PNR > SIR (lag time) > plant yield and N fixation > SIR (respiration after 24 and 48 h) > BR. The lowest adverse effect concentrations were found with the PNR at 7 mg kg(-1) of Cd and 107 mg kg(-1) of Zn. The variability of these endpoints among different uncontaminated soils was additionally assessed on 14 soil samples. That variability showed a strong correlation with sensitivity scores, illustrating that metal-sensitive endpoints have a large natural variability. We question the ecological relevance of highly sensitive microbial assays, because they tend to have a large natural variability. The identification of toxicity in the field requires endpoints that are highly sensitive and that do not vary greatly among soils (i.e., robust); however, no such endpoint was found in the present study. The endpoints that combined average sensitivity and robustness were SIR (lag time), clover yield, and N fixation in clover. PMID- 15779765 TI - Toxicity of sixty-three metals and metalloids to Hyalella azteca at two levels of water hardness. AB - The toxicity of all atomically stable metals in the periodic table, excluding Na, Mg, K, and Ca, was measured in one-week exposures using the freshwater amphipod Hyalella azteca in both Lake Ontario, Canada, and soft water (10% Lake Ontario). Metals were added as atomic absorption standards (63 metals), and also as anion salts for 10 metals. Lethal concentrations resulting in 50% mortality (LC50s) were obtained for 48 of the metals tested; the rest were not toxic at 1,000 microg/L. The most toxic metals on a molar basis were Cd, Ag, Pb, Hg, Cr (anion), and Tl, with nominal LC50s ranging from 5 to 58 nmol/L (1 to 58 nmol/L measured). These metals were followed by U, Co, Os, Se (anion), Pt, Lu, Cu, Ce, Zn, Pr, Ni, and Yb with nominal LC50s ranging from 225 to 1,500 nmol/L (88-1,300 nmol/L measured). Most metals were similarly or slightly more toxic in soft water, but Al, Cr, Ge, Pb, and U were >17-fold more toxic in soft water; Pd was less toxic in soft water. Atomic absorption (AA) standards of As and Se in acid had similar toxicity as anions, Sb was more toxic as the AA standard, and Cr and Mn were more toxic as anions. One-week LC50s for H. azteca correlate strongly with three-week LC50s and three-week effect concentrations resulting in 50% reduction in reproduction (EC50s) in Daphnia magna. PMID- 15779766 TI - Description and initial evaluation of a Xenopus metamorphosis assay for detection of thyroid system-disrupting activities of environmental compounds. AB - A need is recognized for the development and evaluation of bioassays for detection of thyroid system-disrupting compounds. The issue of testing for thyroid disruption can be addressed by exploiting amphibian metamorphosis as a biological model. In the present study, a test protocol for a Xenopus metamorphosis assay (XEMA) was developed and its interlaboratory transferability was evaluated in an informal ring test with six laboratories participating. In the XEMA test, exposure of Xenopus laevis tadpoles was initiated at stages 48 to 50 and continued for 28 d. Development and growth of tadpoles were assessed by means of developmental stage and whole body length determinations, respectively. For initial test protocol evaluation, thyroxine (T4), and propylthiouracil (PTU) were used as positive controls for thyroid system-modulating activity, and ethylenethiourea (ETU) was used as a test compound. Exposure of tadpoles to 1 microg/L T4 produced a significant acceleration of metamorphosis whereas PTU concentrations of 75 and 100 mg/L completely inhibited metamorphosis. Five different ETU concentrations (5, 10, 25, 50, and 100 mg/L) were tested and a concentration-dependent inhibition of metamorphosis was observed. None of the compounds affected tadpole survival, and only PTU caused a slight retardation in tadpole growth. This study demonstrates that the XEMA test provides a sensitive, robust, and practical testing approach for detection of compounds with both agonistic and antagonistic effects on the thyroid system in Xenopus tadpoles. PMID- 15779767 TI - Organochlorine pesticides and mercury in cottonmouths (Agkistrodon piscivorus) from northeastern Texas, USA. AB - Despite their ecological importance and global decline, snakes remain poorly studied in ecotoxicology. In this study, we examined organochlorine (OC) pesticide and mercury accumulation in cottonmouths (Agkistrodon piscivorus) living on a contaminated site in northeastern Texas (USA). Mercury and p,p' dichlorodiphenyldichloroethylene (p,p'-DDE) were detected in all snakes examined. Other OCs, including p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT), methoxychlor, aldrin, and heptachlor, also were detected, but less frequently. Concentrations of p,p'-DDE were higher in fat than in liver, while mercury concentrations were highest in liver, followed by kidney and tail clips. One animal contained the highest mercury concentration yet reported for a snake (8,610 ng/g). Mercury concentrations in liver and kidney were higher in males than females, while no intersex differences in p,p'-DDE concentrations were observed. Concentrations of p,p'-DDE in fat were correlated positively with body size in male cottonmouths but not females, suggesting a slower rate of accumulation in females. Body size strongly predicted mercury concentrations in liver, kidney, and tail clips of both sexes. Tail clips were strong predictors of mercury in liver and kidney in males but not females, suggesting possible sex dependent differences in mercury toxicokinetics. Both long-term field studies and controlled laboratory investigations are needed to adequately assess the response of cottonmouths to chronic contaminant exposure. PMID- 15779768 TI - Squamous epithelial lesion of the mandibles and maxillae of wild mink (Mustela vison) naturally exposed to polychlorinated biphenyls. AB - Approximately 125 km of the Kalamazoo River, located in southwestern Michigan (USA), are designated as a Superfund site, with polychlorinated biphenyls (PCBs) as the contaminant of concern. Mink (Mustela vison) are a naturally occurring predator in this area and also a species of concern because of their known sensitivity to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and structurally similar compounds, such as PCBs. Four of nine mink trapped from the Kalamazoo River area of concern (KRAOC) exhibited histological evidence of a jaw lesion previously identified in ranch mink. The jaw lesion, hyperplasia of squamous epithelium in the mandible and maxilla, is known to be caused by 3,3',4,4',5 pentachlorobiphenyl (PCB 126) and TCDD. Mink trapped from an upstream reference area (Fort Custer Recreation Area [FCRA]) did not exhibit the lesion. Mean concentrations of total PCBs were 2.8 and 2.3 mg/kg wet weight in the livers of mink from the KRAOC and FCRA, respectively, and TCDD toxic equivalent (TEQ) concentrations were 0.30 and 0.11 microg/kg wet weight, respectively. Significant correlations were found between the severity of the lesion and the hepatic concentrations of total PCBs and TEQs. To our knowledge, this is the first published report of the lesion occurring in wild mink. PMID- 15779769 TI - Effects of perchlorate exposure on resting metabolism, peak metabolism, and thyroid function in the prairie vole (Microtus ochrogaster). AB - Perchlorate, the oxidizer component in most solid rocket propellant formulations, is known to inhibit the uptake of iodide into the thyroid gland, thereby reducing production of the thyroid hormones, triiodothyronine and thyroxine (T4). Thyroid hormones regulate metabolism in endothermic organisms and are responsible for maintenance of homeothermic body temperatures. Little is known about the effects of perchlorate on metabolic capacity. The objectives of the present study were to determine if subchronic (51 d; 0, 1, and 10 mg/kg/d) and chronic (180 d; 0.75 mg/kg/d) perchlorate exposure in adult male prairie voles (Microtus ochrogaster) would alter resting metabolic rates as a result of decreased circulating thyroid hormone concentrations and to determine if perchlorate exposure disrupts thermogenesis in mammals exposed to cold stress. Voles exposed to perchlorate for 51 or 180 d experienced no significant alterations in resting metabolic rates at any point during the exposure period. Additionally, the treatment had no effect on peak metabolic rates or plasma thyroid hormone concentrations. However, thyroid gland T4 concentrations were significantly lower in perchlorate-exposed voles than in controls, indicating that thyroid gland T4 content may be a more sensitive endpoint than other thyroid variables for assessing perchlorate exposure. Overall, the present study did not provide evidence for energetic alterations associated with perchlorate exposure at concentrations that are higher than those typically found in groundwater or surface water in the environment. PMID- 15779770 TI - Mercury concentrations in fish from forest harvesting and fire-impacted Canadian Boreal lakes compared using stable isotopes of nitrogen. AB - Total mercury (Hg) concentration was determined in several piscivorous and nonpiscivorous species of fish from 38 drainage lakes with clear-cut, burnt, or undisturbed catchments located in the Canadian Boreal Shield. Mercury concentrations increased with increasing fish trophic position as estimated using stable isotopes of nitrogen (N; r2 = 0.52, 0.49, and 0.30 for cut, reference, and burnt lakes, respectively; p < 0.01). Mercury biomagnification per thousand delta15N varied from 22 to 29% in the three groups of lakes. Mercury availability to organisms at the base of the food chain in lakes with cut catchments was higher than that in reference lakes. In cut lakes, Hg concentrations in fish were significantly related to ratio of the clear-cut area to lake area (or lake volume; r = +0.82 and +0.74, respectively, p < 0.01). Both impact ratios were, in turn, significantly correlated with dissolved organic carbon. These findings suggest that differential loading of organic matter-bound Hg to lakes can affect Hg cycling. In addition, Hg concentrations exceeded the advisory limit for human consumption (0.5 microg/g wet wt) from the World Health Organization in all top predatory species (northern pike, walleye, and burbot) found in cut and in two partially burnt lakes. Thus, high Hg concentrations in fish from forest-harvested and partially burnt lakes may reflect increased exposure to Hg relative to that in lakes not having these watershed disturbances. PMID- 15779771 TI - Is oxidative stress the mechanism of blue sac disease in retene-exposed trout larvae? AB - Retene (7-isopropyl-1-methylphenanthrene) causes blue sac disease (BSD) in early life stages of fish, an effect similar to that of 2,3,7,8 tetrachlorodibenzo(p)dioxin. The signs of BSD include cytochrome P450 (CYP1A) induction, edema, hemorrhaging, and craniofacial deformities, indicating membrane damage, circulatory failure, and impaired development. To test if the underlying cause was oxidative stress, rainbow trout (Oncorhynchus mykiss) larvae were exposed to waterborne retene or to known prooxidants (paraquat, t-butyl hydroperoxide, and carbon tetrachloride) in the presence or absence of vitamin E, an antioxidant. Fish exposed to retene showed an increased prevalence of BSD, reduced tissue concentrations of vitamin E and total glutathione, and a lower percentage of glutathione in a reduced form. Coexposure to vitamin E reduced the prevalence of BSD and restored tissue concentrations of vitamin E, but it did not affect retene uptake or tissue concentrations of glutathione. These responses are consistent with oxidative stress as a mode of action of retene. However, retene did not affect whole-body lipid peroxide concentrations, and prooxidants did not affect the prevalence of BSD and had only minimal effects on tissue glutathione and vitamin E. Possible explanations for these conflicting results include prooxidant exposures were insufficient to generate oxidative stress; lipid peroxidation may not be measurable in whole-body homogenates of retene-exposed fish if effects are localized to endothelial cells, where CYP1A enzymes are most induced; or retene may have an alternate mode of action (e.g., adduction of retene metabolites to lipids, protein, or DNA). PMID- 15779772 TI - Effect of species differences, pollutant concentration, and residence time in soil on the bioaccumulation of 2,2-bis (p-chlorophenyl)-1,1-dichloroethylene by three earthworm species. AB - Laboratory experiments were conducted to study the effects of species differences, soil concentration, and contaminant-residence time in soil on the bioaccumulation factor (BAF; dry-weight ratio of contaminant concentration in the tissue to that in the soil) of 2,2-bis (p-chlorophenyl)- 1,1-dichloroethylene (p,p'-DDE) for three species of earthworms. In four field-weathered soils, the BAF for Eisenia foetida, an epigeic species (surface habitat), was approximately 10-fold higher than those for Lumbricus terrestris, an anecic species (deep habitat) and Aporrectodea caliginosa, an endogeic species (habitat within the soil profile). Preliminary analysis indicates that BAF may decline with increasing pollutant concentration in soil. With regard to contaminant-residence time, the BAF for E. foetida was lower in weathered soils relative to that in freshly amended soils, but age of p,p'-DDE did not significantly alter the BAF for A. caliginosa. These data suggest total chemical concentration alone is not a reliable indicator of the toxicological significance of a contaminated soil and that species-specific differences and environmental factors significantly impact overall exposure and risk. PMID- 15779773 TI - Assimilation and bioconcentration of Ag and Cd by the marine black bream after waterborne and dietary metal exposure. AB - We determined the aqueous uptake and dietary assimilation of Cd and Ag by the marine black bream Acanthopagrus schlegeli following one to four weeks' exposure (or conditioning) to waterborne or dietary Cd or Ag at different concentrations. The concentrations of metals and metallothioneins (MT) in different tissues also were determined. The viscera contained the highest Ag, Cd, and MT concentrations after metal exposure. After exposure to waterborne metals, the metal and MT concentrations in the gills were higher than those in the remaining tissues (mainly muscles and bones), but this pattern was reversed following exposure to dietary metals. The assimilation efficiencies (AEs) of Cd and Ag ranged from 6 to 24% and 15 to 30%, respectively. The rate constant of uptake from the dissolved phase (ku) of Cd and Ag ranged from 2.2 to 7.5 and 8.0 to 31.7 L kg(-1) d(-l), respectively. In all the exposure experiments, the ku and AE increased with induced MT concentration and tissue metal concentration. Increasing metal accumulation may have been due to the increased available binding sites following the induction of MT in the fish. Furthermore, the MT induced by either Cd or Ag was not specific, but was able to bind with both metals and enhance bioaccumulation. Exposure to dissolved and dietary metals may increase metal accumulation, which potentially may lead to metal toxicity, although the fish may develop a tolerance to metals due to the apparent induction of MT. PMID- 15779774 TI - Population genetic structure of a nonmigratory estuarine fish (Fundulus heteroclitus) across a strong gradient of polychlorinated biphenyl contamination. AB - Populations of the estuarine fish Fundulus heteroclitus indigenous to contaminated sites exhibit heritable resistance to some of the toxic effects of early life-stage exposure to polychlorinated biphenyls (PCBs). This evolved tolerance provides evidence of strong selection by PCBs, and it suggests other potential genetic effects of these stressors on resident populations. Environmental contaminants have the potential to affect the genetic structure of populations and to reduce genetic diversity, but species life-history traits, particularly patterns of migration and dispersal, also influence the distribution of genetic variation among populations. Therefore, the present was conducted to determine whether genetic diversity or genetic structure is altered in populations of F. heteroclitus indigenous to 18 sites in Massachusetts (USA) and Rhode Island (USA), representing a steep gradient of sediment PCB contamination and culminating in a Superfund site at New Bedford Harbor (NBH; MA, USA). Allele frequencies at enzymatic loci were used to assess genetic structure and diversity. Selection experiments using a highly toxic PCB congener (3,3',4,4',5 pentachlorobiphenyl) were conducted to determine if genetic patterns at field sites could be associated with contaminant exposures. Although allele frequencies clearly reflected a pattern of isolation by distance, the results indicated neither significant loss of genetic diversity nor alteration of allele frequencies for populations of F. heteroclitus in NBH. PMID- 15779775 TI - Population genetic structure and tolerance to dioxin-like compounds of a migratory marine fish (Menidia menidia) at polychlorinated biphenyl-contaminated and reference sites. AB - The present study was conducted to evaluate evidence of genetic adaptation to local contaminants in populations of the migratory marine fish Menidia menidia residing seasonally in reference sites or an industrial harbor contaminated with dioxin-like compounds (DLCs). For this purpose, we compared DLC sensitivity and genetic patterns of populations sampled from sites both inside and outside New Bedford Harbor (NBH; MA, USA), a U.S. Environmental Protection Agency Superfund site with extreme polychlorinated biphenyl (PCB) contamination. Offspring of M. menidia collected from NBH were significantly less sensitive regarding embryonic exposure to the dioxin-like PCB congener 3,3',4,4',5-pentachlorobiphenyl (PCB 126) than offspring of M. menidia from a reference site. Analysis of 10 polymorphic enzymatic loci indicated little genetic differentiation among populations in the study area. However, genotype frequencies of juveniles from both NBH and an adjacent site in Massachusetts exhibited significant deviations from Hardy-Weinberg equilibrium expectations at one locus, phosphoglucomutase (PGM*). Genetic analysis of survivors of embryonic laboratory exposure to PCB 126 indicated that genotypes at PGM* were related to survivorship. Although a relationship was identified between DLC tolerance and PGM* genotype, regional mixing of M. menidia populations during migration and absence of multigeneration exposure at contaminated sites may limit localized adaptation. PMID- 15779776 TI - Bench-scale evaluation of in situ bioremediation strategies for soil at a former manufactured gas plant site. AB - We examined the biodegradation and desorption of a set of 15 polycyclic aromatic hydrocarbon (PAH) compounds in coal tar-contaminated soil at a former manufactured gas plant site to evaluate the feasibility of in situ bioremediation. Experiments were conducted in well-mixed aerobic soil suspensions containing various additives over a 93- to 106-d period. In general, both biotransformation and desorption decreased with PAH ring size, becoming negligible for the six-ring PAH compounds. Biodegradation by indigenous microorganisms was strongly accelerated by addition of inorganic nutrients (N, P, K, and trace metals). The rates of biotransformation of PAH compounds by indigenous microorganisms in nutrient-amended flasks outpaced their maximum (i.e., chelate-enhanced) rates of desorption to an infinite sink (Tenax) in sterilized systems run in parallel, suggesting that indigenous organisms facilitated desorption. Biodegradation by indigenous organisms in nutrient amended flasks appeared to be unaffected by the addition of a site-derived bacterial enrichment culture, resulting in approximately 100-fold higher aromatic dioxygenase levels, and by the addition of 0.01 M chelating agent (citrate or pyrophosphate), although such chelating agents greatly enhanced desorption in microbially inactivated flasks. The strong ability of nutrients to enhance degradation of the bioavailable PAHs indicates that their persistence for many decades at this site likely results from nutrient-limited natural biodegradation, and it also suggests that an effective strategy for their bioremediation could consist simply of adding inorganic nutrients. PMID- 15779778 TI - Mapping the journey of cancer patients through the health care system. Part 3: An approach to staging. AB - This is the third in a series of articles relating results from a line of research whose intent was to construct a complete history of patient interactions with the health care system using available data sources for all patients diagnosed in 1990 with a primary breast, colorectal, or lung tumour in Manitoba. This article presents details of the development and application of methods to produce TNM staging data on the roughly 2,000 patients in this population. The operational definitions constructed for this research can be adapted for other tumour sites and data sources. Findings include methods developed to overcome the sometimes ambiguous and inconsistent available documentation, which ultimately produced reliable TNM staging data. Survival data for this population by stage of disease are given. PMID- 15779777 TI - Screening test battery for pharmaceuticals in urine and wastewater. AB - A test battery for identifying ecotoxicological hazards was applied to six pharmaceuticals (carbamazepine, diclofenac, ethinylestradiol, ibuprofen, propranolol, and sulfamethoxazole), to their mixtures, and to urine spiked with pharmaceuticals to test the suitability of biotests for screening urine and wastewater and for monitoring the efficiency of wastewater treatment. The test battery comprised the bioluminescence inhibition test with Vibrio fischeri, the yeast estrogen screen, and a photosynthesis inhibition assay in algae based on chlorophyll fluorescence measurements. Mixture and additional experiments with a cocktail of pharmaceuticals added to urine confirmed the applicability of the test systems as an integrated measure of the overall micropollutant burden. Because the concentration of pharmaceuticals in wastewater is low and the nutrients and salts may have a negative impact on the bioassays, urine and wastewater samples were cleaned and concentrated by solid-phase extraction (SPE). The compounds of interest ranged from polar to nonpolar and from positively charged to neutral and negatively charged. Consequently, the SPE method was optimized for universality rather than for specificity. Results of preliminary experiments with raw and treated urine and wastewater indicate the suitability of the proposed test battery for screening urine and wastewater. PMID- 15779779 TI - Do we have the power to create our own future as oncology nurses? PMID- 15779780 TI - Factors influencing the incidence and severity of oral mucositis in patients undergoing autologous stem cell transplantation. AB - This retrospective descriptive study documented the frequency of oral mucositis and examined the impact of certain variables in the development of oral mucositis in autologous stem cell transplants. Oral mucositis occurred in 90% of the patients, but 53.57% developed ulcerations. On average, oral mucositis started on day seven, lasted for six days and resolved on day 13 after the initiation of chemotherapy. Sodium bicarbonate mouthwash and mucositis mouthwash were commonly used interventions; 72.92% of the interventions were used as secondary prevention. Oral mucositis was significantly associated with diagnosis (lymphoma), chemotherapy (etoposide and melphalan), and level of prevention (secondary). Diagnosis (lymphoma), chemotherapy (etoposide and melphalan), serum creatinine (peak level), and level of prevention (secondary) were independent predictors of oral mucositis. PMID- 15779781 TI - What should we measure: infection or infection control? PMID- 15779782 TI - Accountability for health protection in England: how this has been affected by the establishment of the Health Protection Agency. PMID- 15779783 TI - Evaluation of peer review of infection control arrangements for healthcare associated infection in four acute care trusts in the East of England. AB - In order to constructively use Department of Health surveillance data to address the issue of healthcare-associated infection, a peer review process investigating infection control arrangements for healthcare-associated infection in acute care trusts was initiated in the East of England. This study set out to evaluate that process and determine if it should be rolled out regionwide. A questionnaire survey of infection control doctors at trusts visited was conducted by telephone. Respondents commented that the peer review process raised the profile of infection control and healthcare-associated infection in participating trusts, and that a review of routine surveillance data was a useful way to initiate discussion about and focus attention on hospital infection control. Respondents felt the forum for discussion of infection control was most beneficial. A peer review process utilising routine surveillance data is of value in addressing issues related to infection control arrangements for healthcare-associated infection in acute care trusts. PMID- 15779784 TI - Surveillance of antenatal infections--HIV, hepatitis B, syphilis and rubella susceptibility in London. AB - London has relatively high rates of HIV, hepatitis B and syphilis, and has the potential for undiagnosed infection in pregnancy. As part of a drive to improve HIV testing in pregnancy, surveillance of a number of antenatal infections has been carried out in London's 30 maternity units since 2000. Infection screening rates rose from 89% in 2000 to 94% in 2002, and HIV screening rates increased from 66% to 86%. During 2002, 115,230 women booked for antenatal care; 998 were HBsAg positive (about ten in 1000), 452 were HIV positive (five in 1,000), 412 tested positive for syphilis (four in 1000), and 3,929 were susceptible to rubella (36 in 1000). The main burden of infection was concentrated in a few geographic areas. Worryingly the majority of women found to be HIV positive were diagnosed by antenatal screening and not before. PMID- 15779785 TI - Immunisation of babies of women who screen positive for hepatitis B. AB - Pregnant women from countries with high prevalence rates of hepatitis B virus (HBV) infection may have significant HBV DNA levels despite being hepatitis B e antigen (HBeAg) negative and having antibody to HBeAg (anti-HBe positive). It is suggested that their babies, following delivery, should be given hepatitis B specific immunoglobulin (HBIG) in addition to hepatitis B vaccine in order to ensure their protection against infection. PMID- 15779786 TI - General practitioners' concerns about childhood immunisation and suggestions for improving professional support and vaccine uptake. AB - In recent years childhood vaccination has been vigorously debated. Professional and parental confidence in the measles, mumps and rubella (MMR) vaccine in particular has been shaken, as reflected by its decreased uptake. A semi structured postal questionnaire survey of general practitioners (GPs) working in the Highland region of Scotland was undertaken to assess their views on vaccination issues. A 73% response rate was achieved. Of respondents 28% expressed concerns about the possible side effects of MMR. Of GPs 98% thought it was 'likely' that the benefits of the DTP-Hib, polio and men C vaccines outweighed the possible risks. For the first and second doses of MMR 92% and 86%, respectively, believed this to be true. Approximately three quarters of GPs described themselves 'very confident' in discussing the DTP-Hib vaccine with parents, compared with 57% for MMR. Some respondents were misinformed regarding the adverse events associated with the MMR vaccine. There is professional concern about MMR. Initiatives to support GPs in promoting vaccination in order to improve immunisation rates are discussed. PMID- 15779787 TI - Survey of local immunisation training in England--the case for setting national standards. AB - In June 2003 a questionnaire on immunisation training was sent to the 302 primary care trusts (PCTs) in England to ascertain the frequency and content of immunisation training being offered to healthcare professionals. Fifty-four per cent of the 227 trusts (75%) who replied were concerned about their ability to deliver an immunisation programme. Contributing factors included the lack of a designated training lead, shortage of specialist input, available time, and funding. Of PCTs, 33/219 (15%) were not providing immunisation training sessions for practice nurses, 28/219 (13%) for health visitors, and 30/219 (14%) for school nurses; 67/219 (31%) had no sessions organised for general practitioners. Most 138/175 (79%) PCTs would welcome the introduction of some national minimum standards for immunisation training to assist them in setting up and maintaining a programme, and allocating sufficient resources to it. PMID- 15779788 TI - Outbreak of influenza A (H3N2) in a highly-vaccinated religious community: a retrospective cohort study. AB - An outbreak of influenza occurred at the end of the 2001-2 winter season in a highly-vaccinated, semi-enclosed, religious community. On the basis of retrospective self reporting, 43% met the case definition (151/350) and 81% (25/31) of throat swabs from cases were polymerase chain reaction (PCR) -positive for influenza A (H3N2). The risk of developing influenza in people aged 65 or more was lower than that of children aged 2 years and under (odds ratio 0.1, 95% confidence interval 0.02 to 0.38). The risk of developing symptoms of influenza was not significantly different between people who had been vaccinated in the United Kingdom and those who had not been vaccinated (OR 1.14, CI 0.41 to 3.14). Chronic disease was an independent risk factor for developing symptoms of influenza (OR 1.9, CI 1 to 3.63). Timing of the outbreak, the age structure and mode of communal living may have influenced the efficacy of the influenza vaccine, which was well matched to circulating strains at the time of the outbreak. It is important to consider the diagnosis of influenza even in a highly vaccinated community. PMID- 15779789 TI - Acceptability of universal hepatitis B vaccination among school pupils and parents. AB - The World Health Organisation (WHO) recommends universal hepatitis B (hepB) vaccination for all countries, but this policy has not been adopted in the UK and its acceptability there is unknown. We investigated the attitudes of secondary one (S1) school pupils aged 12-13 years (n = 50) and parents (n = 39) using semi structured focus group discussions. There was a lack of awareness of hepB among most participants prior to the study. Parents sought further information, including the risks of infection and vaccine side effects. No participants identified cultural or socioeconomic barriers to being vaccinated against hepB. The majority of pupils and nearly all parents were in favour of universal hepB vaccination. Offering hepB vaccination to all S1 pupils, in school, should therefore be highly acceptable, providing that sufficient information on the risk of hepB infection and vaccine safety is provided. A facility for answering questions and a forum for pupil education should also be offered. PMID- 15779790 TI - Tetanus immunisation policy in England and Wales--an overview of the literature. AB - Current UK policy on immunisation for tetanus has changed in the light of evidence that five doses of vaccine probably provides sufficient protection as long as high-risk wounds are managed with tetanus immunoglobulin. This paper reviews the evidence base for tetanus immunisation policy in England and Wales: the epidemiology of tetanus, vaccination coverage and response to tetanus toxoid, and population immunity to tetanus. The paper highlights gaps in our current knowledge of tetanus vaccination and policy implementation, and makes recommendations for further investigations. PMID- 15779791 TI - Sharps discarded in inner city parks and playgrounds--risk of bloodborne virus exposure. AB - The objectives of this study were to determine the number of discarded syringes found in four parks in Southwark, South London, over a specific time period and to test their contents for the presence of hepatitis B virus (HBV) and hepatitis C virus (HCV). Of 106 syringes collected over a four-month period, evidence of HBV was detected in 4.7% (5/106) and HCV in 4.7% (5/106). Urban children, park users and workers are at risk of contact with sharps which may be contaminated with both viruses. Park users need more information on what to do in the event of an injury, and park workers should be immunised against HBV and educated on safe disposal of sharps. PMID- 15779792 TI - Hepatitis A vaccination--a prison-based solution for a community-based outbreak? AB - In December 2001, an increase in cases of hepatitis A was observed in South Yorkshire. Cases were predominantly young males who reported injecting drug use. A community-based vaccination programme was introduced in November 2002, but new cases continued to occur. In March 2003, a vaccination campaign was implemented in the local prison for a four-week period. One thousand two hundred and thirty six (91%) prisoners were vaccinated. Two thirds (895/1,363) of the prisoners came from the area affected by the outbreak and 52% (465/895) reported injecting drugs. The median age of injectors was 25 years. Notifications of cases of hepatitis A from South Yorkshire ceased in August 2003. Although on this occasion the prison vaccination campaign was probably implemented too late to have had a significant impact on the local outbreak, a large number of young male injectors from the local area were successfully vaccinated. This suggests that a prison based intervention offers a potentially effective way of immunising the IDU population and interrupting a community-based outbreak. PMID- 15779793 TI - Acceptance and completion of hepatitis B vaccination among drug users in New York City. AB - Hepatitis B (HBV) vaccination rates remain low among drug users. We examined correlates of vaccine acceptance and completion in two ongoing prospective studies of young injecting and non-injecting drug users in New York City. Street recruited drug users were enrolled at one of two neighbourhood locations (Harlem and the Bronx) between 2000 and 2004 and completed risk behaviour questionnaires and HBV testing. Free HBV vaccination was offered. Among 1117 participants, 26.1% (275) had a previous HBV infection, 57.9% (610) were susceptible to HBV, and 16.0% (169) had serological evidence of previous vaccination. Of the 610 participants susceptible to HBV, 466 (76.4%) returned for their results and were offered vaccination; 53.9% (251) received at least one dose of the vaccine (acceptors). Correlates of vaccine acceptance included older age, public assistance as main income source, and being recruited in the Bronx. Daily crack users were significantly less likely to initiate the vaccine series. Among 240 vaccine acceptors, 98 (40.8%) completed all three doses. Daily injectors, Hispanics, and those recruited in Harlem were less likely to complete the vaccination series. HBV vaccination acceptance among drug users seems likely in programmes that are convenient and offer remuneration; however, extended efforts are needed to improve series completion. PMID- 15779794 TI - Audit of bloodborne virus prevention activity with drug users seen in primary care. AB - Drug misuse is increasingly being managed in general practice. It has been proposed that better use could be made of this contact to identify people with bloodborne virus (BBV) morbidity and to deliver prevention strategies. The Hepatitis B and C Prevention Project was designed to enhance the work of primary healthcare teams in preventing transmission of BBVs in people known to have a history of problem drug use. As part of this work a baseline audit of current provision was undertaken and the results are reported here. Primary care records of 1278 people with a history of illicit drug use were audited to establish the levels of hepatitis B immunisation and testing for BBVs and to determine whether there was a record of any professional discussion of BBV issues with the patient. Records were drawn from rural and city-based general practices. Audit feedback, training, and advice were offered to raise awareness and discussion of how this work was currently being undertaken, and how it might be improved. This baseline audit showed that 90% (n = 1153) of the patients had been questioned about injecting drug use and of these 50% (579/1153) reported injecting at some point in the past. Only 4% (54/1278) had completed a course of hepatitis B immunisation and of these three quarters gave a history of injecting drug use. Another 6% (74/1278) of patients tested for hepatitis B virus (HBV) showed markers of natural immunity. Up to 90% of this group therefore remained vulnerable to this preventable disease. A discussion of BBV issues with a professional was recorded in41% (523/1278) of cases, and was more likely to have occurred in those with a known history of injecting. Individuals were less likely to have been tested for hepatitis C virus (HCV) than for HIV or HBV despite its high prevalence in this group. Only 28% (354/1278) were tested for HCV compared with 33% (416/1278) tested for HBV and 36% (454/1278) tested for HIV. Prevalence of anti-HCV for people with a history of injecting was 51% (137/268) compared to 11% (9/83) in those with no history of injecting. Prevalence of anti-HIV in those with a history of injecting was 10% (29/294) compared 0.7% (1/137) in those with no history of injecting. Prevalence of HBV markers in those with a history of injecting was 23% (65/279) compared with 7% (8/114) in those with no history of injecting. Of the 530 patients with test results, only 52% (275/530) had been tested for all three viruses despite the common transmission routes. PMID- 15779795 TI - Increasing hepatitis B vaccine coverage in prisons in England and Wales. AB - The most frequently reported risk factor for hepatitis B infection in England and Wales is injecting drug use (38%). Since approximately 61% of injecting drug users (IDUs) had been imprisoned and less than 40% had received hepatitis B vaccine, a prison based hepatitis B vaccination programme was set up in 2001. At the 42 establishments participating in this study, all prisoners were offered vaccine at reception. Prisoners over 18 years were vaccinated using the 0, 7 and 21 days schedule and those under 18 years, using the 0, 1 and 2 months schedule. As far as possible a fourth dose was given to all after 12 months. In 2003, 14,163 prisoners received at least one dose of vaccine and altogether 26,265 doses were administered. A further 1111 prisoners reported they had already been vaccinated against hepatitis B. The median vaccine coverage rate was 17% (range 0 94%). Despite low coverage levels, the vaccination programme in prisons can be said to have vaccinated a sizable number of young, male prisoners, a group that have previously been shown to be at high risk of infection. The prisons which achieved vaccine coverage levels over 50% had designated nursing staff who ran the vaccination clinics. PMID- 15779796 TI - Importance of using an HIV Ag/Ab combined assay in a UK population at high risk of acquiring HIV infection. AB - The combined assays for HIV p24 antigen and antibody (4th generation) have had a mixed reception in testing laboratories. A series of five cases detectable only by 4th generation assays or RNA viral load and negative in a variety of 3rd generation assays has prompted us to change our 'routine' screen to a combined assay. PMID- 15779797 TI - Enhanced surveillance of HIV infection in the West Midlands. AB - The West Midlands Regional HIV Surveillance Project was set up in 1991 in response to a need for enhanced surveillance of HIV at a local level for service planning. The system showed that accurate and timely data could be collected whilst ensuring patient confidentiality. We present some of the data obtained from the system. HIV diagnoses continue to increase year on year, with certain population subgroups, including men who have sex with men and black Africans, disproportionately affected. PMID- 15779798 TI - Relationship between the use of hospital services and deprivation score of place of residence among HIV-positive individuals in the north west of England. AB - We used routine surveillance data to investigate whether deprivation relates to hospital admission in a HIV-positive population. HIV-positive individuals living in the poorest areas were more likely to have spent one or more nights in hospital for HIV-related care (adjusted odds ratio = 1.6, p = 0.009, after controlling for infection route, disease stage and demographic variables). This implies that healthcare networks in poorer areas may incur disproportionately greater costs. PMID- 15779799 TI - Clostridium difficile: a questionnaire survey of laboratory practice in England, Wales, and Northern Ireland. AB - Since January 2004, the incidence of Clostridium difficile associated disease (CDAD) has been monitored by a systematic, national, laboratory surveillance system. This system incorporates the recommendations of a body of experts, the National Clostridium difficile Standards Group, which was convened in 2002 to advise the Department of Health (DoH). The recommendations of the group were informed by a questionnaire survey of current practice, and the results of that survey have been used to assess the implications of the recommendations on laboratory practice. Large variability was found as to the specimens selected, tested, and reported on for C. difficile. Standardisation of the diagnosis and reporting of CDAD is desirable and necessary to increase understanding of its epidemiology. PMID- 15779800 TI - Evaluation of Binax NOW RSV as an acute point-of-care screening test in a paediatric accident and emergency unit. AB - Point-of-care tests (POCTs) for respiratory syncytial virus infections are a useful adjunct to reduce the risk of healthcare-associated infections in paediatric wards. A new test based on immunochromatography, Binax NOW RSV, was introduced in the winter of 2002-03. It has user friendly features making it particularly suitable for non-laboratory personnel in a paediatric accident and emergency unit. A prospective study comparing the POCT with laboratory-based direct immunofluorescence (DIF) showed sensitivity of 87% specificity of 94%, positive predictive value of 89% and negative predictive value 92%. The simplicity of Binax NOW RSV should not detract from training staff and maintaining consistent vigilance on quality control. PMID- 15779801 TI - Audit of the detection and notification of meningococcal disease at a North London university district hospital. AB - The microbiology department at a North London hospital audited notifications of suspected meningococcal disease in response to a reported high rate, which it suspected was due to over-reporting out of hours to public health specialists. All cases notified over a 14-month period were assessed with regards to time of reporting and subsequent microbiological confirmation. Of 36 notifications, 18 were confirmed as bacterial, mostly meningococcal meningitis. Of the four types of sample readily obtainable on admission (blood culture, ethylene diamine tetra acetic acid [EDTA] blood sample, throat swab and acute serum) all were obtained in only two cases. The reporting was considered appropriate, reflecting the relatively high incidence of meningococcal disease (MD) within the local population. Further cases might perhaps have been laboratory confirmed had all samples been sent on admission. A single form requesting all four samples in suspected meningococcal disease cases is recommended. Informing public health specialists out of hours could be rationalised. PMID- 15779802 TI - How complete and accurate is meningococcal disease notification? AB - Effective public health control of meningococcal disease (meningococcal meningitis and septicaemia) is dependent on complete, accurate and speedy notification. Using capture-recapture techniques this study assesses the completeness, accuracy and timeliness of meningococcal notification in a health authority. The completeness of meningococcal disease notification was 94.8% (95% confidence interval 93.2% to 96.2%); 91.2% of cases in 2001 were notified within 24 hours of diagnosis, but 28.0% of notifications in 2001 were false positives. Clinical staff need to be aware of the public health implications of a notification of meningococcal disease, and of failure of, or delay in notification. Incomplete or delayed notification not only leads to inaccurate data collection but also means that important public health measures may not be taken. A clinical diagnosis of meningococcal disease should be carefully considered between the clinician and the consultant in communicable disease control (CCDC). Otherwise, prophylaxis may be given unnecessarily, disease incidence inflated, and the benefits of control measures underestimated. Consultants in communicable disease control (CCDCs), in conjunction with clinical staff, should de-notify meningococcal disease if the diagnosis changes. PMID- 15779803 TI - Outbreak of a new Salmonella phage type in South West England: alternative epidemiological investigations are needed. AB - An outbreak of Salmonella Typhimurium DT193a occurred in South West England in April-May 2003. Preliminary investigation suggested an association with the consumption of ham. A matched case-control study was implemented, together with an environmental investigation and a geographic information system (GIS) analysis. Thirty-seven cases and 38 controls were enrolled in the study. Matched analysis was based only on 23 cases, as the other cases did not nominate controls. Eighty per cent of cases and 51% of controls had eaten ham (odds ratio = 3.5, p = 0.03). Cases did not differ from controls in terms of distance from outlets providing ham. All environmental samples collected were negative. The epidemiological evidence indicated an association between this outbreak and consumption of ham, but the environmental investigation was inconclusive. Alternative methods should be considered in support of traditional epidemiological investigation. PMID- 15779804 TI - Susceptibility to infectious rash illness in pregnant women from diverse geographical regions. AB - Varicella-zoster (VZV), rubella (RV) and parvovirus B19 (B19V) infections are important causes of rash illness in pregnancy, due to their potential adverse impact on both mother and fetus. We determined susceptibility to these infections in pregnant women attending our hospital in 2002. Age and nationality were recorded. Sera were tested for VZV, RV, and B19V antibody by enzyme immunoassay. Of 7,980 women screened for VZV IgG, 11.3% were seronegative and therefore susceptible to infection. Across different worldwide regions, 6.9% of Irish and other Western European women were susceptible to VZV, compared to 19.7% of other women tested (p < 0.001), most of whom were from Central and Eastern Europe, sub Saharan Africa and Asia. Of 7,872 women screened for RV IgG, 2.3% were seronegative. Few Irish (0.6%) or other Western European women (0.7%) were rubella non-immune, but 5.5% of women from other regions tested were susceptible to rubella (p < 0.001). A random subset of 1,048 women were tested for B19V IgG. About 38% were susceptible, varying from 22% to 63% across the different regions studied. There are important differences in immunity to these infections and so of potential risk of an adverse outcome in indigenous and immigrant pregnant women in Ireland. PMID- 15779805 TI - Airborne Campylobacter infection in a poultry worker: case report and review of the literature. AB - A man who had recently started work as a packer in a chicken factory developed campylobacteriosis with severe complications. This prompted a qualitative assessment of the occupational infection risk. It is likely that his infection occurred by droplet transmission via the mouth. Serological studies have shown increased risk of infection to poultry workers, particularly in the first weeks of employment. Previous reports have identified the risks of airborne pathogen transmission, and these papers are reviewed here. Epidemiological evidence from the plant indicated that workers were three times more likely than the general population to suffer campylobacteriosis, and occupational health reporting confirms the risk to poultry workers. Employers should offer face masks to their workers for protection from airborne infection. PMID- 15779806 TI - Assessment of HIV-2 testing practices and reporting in England, Wales, and Northern Ireland. PMID- 15779807 TI - Selective neonatal BCG immunisation in a multiethnic community: poor uptake among white British infants. PMID- 15779808 TI - MMR debate: how many children are actually receiving single vaccines? PMID- 15779809 TI - Biotechnology and microbiology--have reached the end of the line? PMID- 15779810 TI - Issues in career progression in microbiology. AB - We are three highly motivated university graduates who are looking to begin careers in microbiology. We each attained a degree in a biological science discipline, one in biochemistry, one in microbial and cellular biology and one in biomedical sciences (the last gained in South Africa). However, the career obstacles we have so far encountered require us to be Olympic hurdlers. This article highlights the issues, and the advantages and disadvantages of the options open to us. We look at each career option in turn and describe the difficulties encountered. PMID- 15779812 TI - [Klippel-Trenaunay syndrome]. AB - The author in connection with three properly observed female patients summarizes the clinical entity of Klippel-Trenaunay syndrome, its partly resembling partly dissociating features facing to other vascular congenital anomalies, and the most important aspects of vasculogenesis. Based on the observation, attention is distinctly called on the first case in whom the Klippel-Trenaunay syndrome presenting with multiple mesenchymal anomalies was associated with selective congenital IgA deficiency. The second was the daughter of this first patient, who also had the syndrome and a selective congenital IgA deficiency. The other daughter of the patient was free from both of the pathological conditions. The third case represented the features of the typical syndrome without verified immunodeficiency. The literary data do not speak about possible correlations between Klippel-Trenaunay syndrome and immunodeficiencies. The pathogenesis of the process and of its possible connections with immunodeficiencies remain to be investigated. PMID- 15779811 TI - [Chronic myeloid leukemia]. AB - Chronic myeloid leukemia is still the most characteristic entity of the chronic myeloproliferative diseases. The tumor promoter role of able-dependent tyrosine kinase activation, which is enhanced by bcr/abl rearrangement (due the classical translocation of Philadelphia chromosomal abnormality) has been quite well clarified. The better understanding of the role of altered cell signalling pathways in the pathogenesis of chronic myeloid leukaemia opened new areas for extensive and fruitful pharmacological research. The first, non-myelosuppressive agent, which was able to reduce the number of Philadelphia positive clonal cells was the interferon group, which drug could substantially prolong the chronic phase and mortality of chronic myeloid leukaemia. Imatinib mesylate, a tyrosine kinase inhibitor seems to be able to produce clinical and major cytogenetic response in more than 80% of patients. Imatinib is also a powerful agent in the accelerated or blastic phased of chronic myeloid leukaemia. With the advent of these new drugs the therapeutic algorithm of chronic myeloid leukaemia and allogenous bone marrow transplantation seems to be reconsidered, too. PMID- 15779813 TI - [Results of photorefractive keratectomy in myopia with flying spot excimer laser with high speed and small beam diameter]. AB - PURPOSE: To examine the result of photorefractive keratectomy (PRK) with the next generation of flying spot excimer laser for the treatment of myopic refractive errors. PATIENTS AND METHODS: During the study 164 eyes of 98 patients were assessed. The average age was 29.4 +/- 4.62 years. The myopic eyes were divided into 3 groups: in Group 1. (n = 115 eyes) the eyes between -1.0 D and -6.0 D were evaluated, in this group the preoperative average refraction was -3.43 +/- 1.2 D (SE = spherical equivalent); in Group 2. (n = 31 eyes) the eyes between -6.1 D and -9.0 D were evaluated; the preoperative average refraction was -7.26 +/- 2.4 D (SE). In Group 3. (n = 18 eyes) eyes above -9.0 D were examined; in Group 3. the average preoperative refraction was -10.22 +/- 3.4 D (SE) Follow-up is 6 months. PRK treatments were carried out with the Zeiss Meditec MEL 80 G-Scan flying spot excimer laser, which was operated with a 0.7 mm beam diameter and 250 Hz frequency. RESULTS: In Group 1. The preoperative correction decreased to -0.14 D (SE); in 71% of the treated eyes uncorrected visual acuity (UCVA) was 1.0; in 24% of the eyes UCVA improved to 1.2. Best corrected visual acuity (BCVA) was unchanged in 65% of the eyes; while in 34 improved (in 24% it improved by more than 2) Snellen lines. In Group 2. the preoperative correction decreased to -0.13 +/- 0.01 D (SE); 67% had 1.0 and 8% had a 1.2 UCVA. BCVA was unchanged in 92% in Group 2.; in 8% BCVA increased by 1 or 2 Snellen lines. In Group 3., the preoperative correction decreased to -0.44 +/- 0.2 D (SE); 96% of the eyes had a 0.8 UCVA; BCVA did not change in 75% of the eyes, 25% of the eyes gained 1 Snellen line. CONCLUSIONS: Myopic PRK treatments with the 3rd generation of excimer laser were effective and safe. Regarding BCVA, best results were obtained in the low and medium myopia groups. PMID- 15779814 TI - [Ethical aspects of disclosing information on prenatal screening for Down's syndrome]. AB - Giving detailed information on prenatal screening for Down's syndrome is considered as paramount since this medical procedure intends to enhance the patient's self-governance in reproductive issues. Not only the respect for autonomy, but also the increased maternal anxiety and the reproductive decisions following the positive test result demand from the genetic professional to offer the test through genetic counselling. The counsellor's awareness about the expectations of pregnant women and the clarification of her own attitude concerning the screening can contribute to the effectiveness of counselling. The content of information embraces the technical aspects of screening and its consequences, like the description of Down's syndrome, the method of screening, the way of risk assessment, the detection rate, the false positive and false negative test results, the diagnostic procedures, and the termination of pregnancy. Written information leaflets should be completed by personal communication as the combination of these two forms has proved to be the most useful. The process of consultation is influenced by the communication skill of the genetic professional and the information seeking activity of the patient, so doctors should be trained to communicate better and patients should be encouraged to get more information about the screening. PMID- 15779815 TI - [Practical guide-lines for the treatment of hypertension]. PMID- 15779816 TI - [Unveiling a Semmelweis statue]. PMID- 15779818 TI - Desperate to stop the flow of red ink, level I trauma center will deny transfers. AB - Despite financial hardship, the Emergency Medical Treatment & Labor Act (EMTALA) may place severe limitations on solutions available to overcrowded EDs. --It is not up to the receiving hospital to determine capabilities of the transferring facility. --If you suspect an improper transfer request, accept the transfer and then file a complaint with the Centers for Medicare & Medicaid Services. - Whistle-blower laws protect ED managers if they refuse or complain about orders that violate EMTALA. PMID- 15779817 TI - [Remembrance of Lajos Barta (1913-2000)]. PMID- 15779819 TI - New report highlights crisis with on-call panels. AB - To solve the challenge of filling call panels, money, turf, and lifestyle issues of physicians must be addressed. --Use of hospitalists and physician assistants can help provide coverage for internal medicine cases. --Competitive contracting could create a critical mass of available on-call physicians. --Paying physicians to be on call makes sense, but adequate funding must be provided. PMID- 15779820 TI - To ease overcrowding, delay elective surgeries. AB - Make sure you are in the loop when decisions about postponing surgeries are made. --The ED managers often have the most accurate information available concerning likely demand for beds. --Try to be part of the drafting process for any new policies. --A response plan that does not include ED input is likely to suffer a breakdown. PMID- 15779821 TI - 'Predicting the future' helps cut LOS by 50%. AB - Being able to predict demand is critical to achieving proper staffing levels and optimal patient flow. --No need to invest in fancy software; a basic Excel program can perform the functions you require. --If you don't integrate your data with the rest of the hospital, you actually may make things worse. PMID- 15779822 TI - Most ED patients feel safe, but many fear errors. AB - Involving your patients in the care process can help ease their fears and promote safety. --In a culture of safety, ED managers must be accountable for keeping patients well informed. --Easing patient safety concerns can contribute to higher levels of satisfaction and more return visits. --Even worries about potential errors can affect the likelihood a patient will recommend your facility. PMID- 15779823 TI - Proton pumping inorganic pyrophosphatase of endoplasmic reticulum-enriched vesicles from etiolated mung bean seedlings. AB - Endoplasmic reticulum (ER)-enriched vesicles from etiolated hypocotyls of mung bean seedlings (Vigna radiata) were successfully isolated using Ficoll gradient and two-phase (polyethylene glycol-dextran) partition. The ER-enriched vesicles contained inorganic pyrophosphate (PPi) hydrolysis and its associated proton translocating activities. Antiserum prepared against vacuolar H+-pyrophosphatase (V-PPase, EC 3.6.1.1) did not inhibit this novel pyrophosphatase-dependent proton translocation, excluding the possible contamination of tonoplast vesicles in the ER-enriched membrane preparation. The optimal ratios of Mg2+/PPi (inorganic pyrophosphate) for enzymatic activity and PPi-dependent proton translocation of ER-enriched vesicles were higher than those of vacuolar membranes. The PPi dependent proton translocation of ER-enriched vesicles absolutely required the presence of monovalent cations with preference for K+, but could be inhibited by a common PPase inhibitor, F-. Furthermore, ER H+-pyrophosphatase exhibited some similarities and differences to vacuolar H+-PPases in cofactor/substrate ratios, pH profile, and concentration dependence of F-, imidodiphosphate (a PPi analogue), and various chemical modifiers. These results suggest that ER-enriched vesicles contain a novel type of proton-translocating PPase distinct from that of tonoplast from higher plants. PMID- 15779824 TI - Uptake, transport and accumulation of nicotine by the Golden Potho (Epipremnum aureum): the central role of root pressure. AB - The roots of Epipremnum aureum, though not synthesizing nicotine themselves, take up exogenously fed nicotine as a xenobiotic. The alkaloid is subsequently translocated to the leaves, via the xylem path, where it accumulates in the mesophyll up to levels comparable with nicotine-rich Nicotiana species. The Epipremnum plants accept nicotine only up to a distinct level; saturation is reached after about 10 days. All mature, non-senescent leaves accumulate the same amount of nicotine. By different experimental approaches, unequivocal evidence could be provided that root pressure is the 'translocative force' for nicotine transport in E. aureum. Xylem sap exudates, collected from shoot stumps that were connected to an intact root system immersed in nicotine solution were analyzed for nicotine content. Nicotine uptake from the medium by the root and its subsequent transfer into the xylem of the shoot persisted for more than 10h without measurable decline of the transport rate, provided the nicotine concentrations applied were < or =0.05%. In intact plants, where both components of water transport in the xylem--root pressure and transpirative water flow--are in operation, no surplus transport of nicotine from the roots into the leaves took place beyond the level observed in amputated plants. Under the influence of inhibitors of root respiration, nicotine uptake was halted slowly in case of oxygen deprivation and in case of cyanide, or it stopped very rapidly when CCCP, an uncoupler of mitochondrial ATP formation, was applied to the roots. This threshold of toxicity against the xenobiotic was established by dose effect curves for nicotine sensitivity of the roots for root respiration and by transpiration measurements. Leaves, bearing a heavy 'nicotine load', showed symptoms of senescence only after 3-6 weeks, as indicated by a decline in the chlorophyll content, the chl a/b ratio, and the maximal quantum yield efficiency (Fv/Fm), and by an increase in catalase activity. Our results provide insight into the mechanisms of uptake, transport and storage of nicotine as a xenobiotic. PMID- 15779825 TI - Apple flavonols during fruit adaptation to solar radiation: spectral features and technique for non-destructive assessment. AB - Spectral properties of flavonols of three varieties (Golden Delicious, Antonovka, and Renet Simirenko) of anthocyanin-free apple fruit were investigated with reflectance spectroscopy. The results of spectral and biochemical analyses suggested that fruit reflectance in a broad spectral range 365-430 nm is strongly dependent on and, in sunlit fruit surfaces, governed by flavonols. The build up of peel flavonols (mainly rutin and other quercetin glycosides) resulted in a dramatic decrease of fruit reflectance in this range, flattening of the spectrum, and extending the region with low reflectance (4-5%) to ca. 410 nm. The spectral features observed suggest that flavonols contribute significantly to screening of excessive radiation, not only UV-A, but in the short-wave bands of chlorophyll and carotenoid absorption in the visible part of the spectrum as well. To retrieve quantitatively flavonol content from reflectance spectra, we tested the applicability of an inversion technique developed for non-destructive leaf pigment assessment. The model for flavonol content assessment was suggested in the form (R(-1)410 - R(-1)460)R800, where Rlambda is reflectance at wavelength lambda. The model was linearly related to flavonol content between 8 and 220nmol/cm2 with the coefficient of determination r2=0.92 and root mean square error of flavonol estimation of 20 nmol/ cm2 regardless of cultivar, chlorophyll, and carotenoid content. PMID- 15779826 TI - The variable C-terminus of 14-3-3 proteins mediates isoform-specific interaction with sucrose-phosphate synthase in the yeast two-hybrid system. AB - Sucrose-6-phosphate synthase (SPS) is a target for 14-3-3 protein binding in plants. Because several isoforms of the 14-3-3 protein are expressed in plants, I investigated which isoforms have the ability to bind SPS. Two 14-3-3 isoforms (T14-3d and a novel isoform designated T14-3 g) were found to interact with SPS from tobacco (Nicotiana tabacum L.) in a two-hybrid screen. To further address the question of isoform specificity of 14-3-3s, four additional isoforms were tested for their ability to interact with SPS in the yeast two-hybrid system. The results clearly revealed large differences in affinity between individual 14-3-3 isoforms toward SPS. Deletion analysis suggested that these differences were mediated by the variable C-terminus of 14-3-3s. Site-directed mutagenesis of candidate 14-3-3 binding sites on SPS demonstrated that interaction could be independent of a phosphorylated serine residue within conserved binding motifs in the yeast system. These findings suggest that the large number of 14-3-3 isoforms present in plants reflects functional specificity. PMID- 15779827 TI - Rapid degradation of starch in chloroplasts and concomitant accumulation of soluble sugars associated with ABA-induced freezing tolerance in the moss Physcomitrella patens. AB - Abscisic acid (ABA) has been postulated to play a role in the development of freezing tolerance during the cold acclimation process in higher plants, but its role in cold tolerance in tower land plants has not been elucidated. The moss Physcomitrella patens rapidly developed freezing tolerance when its protonemata were grown in a medium containing ABA, with dramatic changes in the LT50 value from -2 degrees C to over -10 degrees C. We examined physiological and morphological alterations in protonema cells caused by ABA treatment to elucidate early cellular events responsible for rapid enhancement of freezing tolerance. Microscopic observations revealed that ABA treatment for 1 day resulted in a dramatic alteration in the appearance of intracellular organelles. ABA-treated cells had slender chloroplasts, with a reduced amount of starch grains, in comparison with those of non-treated cells. The ABA-treated cells also had several segmented vacuoles while many of non-treated cells had one central vacuole. When frozen to -4 degrees C, freezing injury-associated ultrastructural changes such as formation of aparticulate domains and fracture-jump lesions were frequently observed in the plasma membrane of non-treated protonema cells but not in that of ABA-treated cells. The ABA treatment increased the osmotic concentration of the protonema cells, in correlation with accumulation of free soluble sugars. These results suggest that ABA-induced accumulation of soluble sugars, associated with morphological changes in organelles, mitigated freezing induced structural damage in the plasma membrane, eventually leading to enhancement of freezing tolerance in the protonema cells. PMID- 15779828 TI - Biophysical studies of photosystem II-related recovery processes after a heat pulse in barley seedlings (Hordeum vulgare L.). AB - Leaves of 7-day-old barley seedlings were subjected to heat pulses at 50 degrees C for 20 or 40s to inhibit partially or fully the oxygen evolution without inducing visible symptoms. By means of biophysical techniques, we investigated the time course and mechanism of photosystem II (PSII) recovery. After the heat treatment, the samples were characterized by typical heat stress symptoms: loss of oxygen evolution activity, strong decrease of Fv/Fm, induction of the K-step in the fluorescence induction transient, emergence of the AT-thermoluminescence band and a dramatic increase in membrane permeability. In the first 4h in the light following the heat pulse, the AT-band and the K-step disappeared in parallel, indicating the loss of this restricted activity of PSII. This phase was followed by a recovery period, during which PSII-activity was gradually restored in the light. In darkness, no recovery, except for the membrane permeability, was observed. A model is presented that accounts for (i) the damage induced by the heat pulse on the membrane architecture and on the PSII donor side, (ii) the light-dependent removal of the impaired reaction centers from the disorganized membrane, and (iii) the subsequent light-independent restoration of the membrane permeability and the de novo synthesis of the PSII reaction centers in the light. PMID- 15779829 TI - The effect of ozone on the yellowing process of magnesium-deficient clonal Norway spruce grown under defined conditions. AB - During two vegetation periods, young clonal spruce trees (Picea abies (L.) Karst.) with sufficient and poor magnesium (Mg) supply were exposed in the environmental chambers of the GSF phytotron to three levels of ozone (daily means: 18-22, 88-130, and 135-190 microg m(-3); 10% reduction at night). Previous year's needles were examined at 4-week intervals with respect to their contents of Mg, Ca, K, Mn, N, P, and chlorophyll (Chl), various parameters of Chl fluorescence, and the stability of the isolated light-harvesting Chl-a/b-protein complex LHC II. The needles of the two nutrition variants contained more than 0.53 or less than 0.27mg Mg g(-1) needle dry matter, respectively. The ratio of variable to maximal Chl-a fluorescence of the dark-adapted needles, Fv/Fm, and the photoinhibitory quenching of Fv after light treatment, SVi.v, were affected by the Mg content of the needles rather than the ozone levels. Changes of the Chl content and the behavior of the LHC II allowed differentiating between a slow process of needle yellowing occurring under Mg deficiency only, and a rapid process of needle yellowing occurring under the combined action of Mg deficiency and ozone pollution. Only the rapid yellowing process was accompanied by destabilization of the LHC II, and the degree of destabilization was correlated with the ozone concentration present in the days before sampling. The results are consistent with observations obtained at a research site in the Central Black Forest (J Plant Physiol 161 (2004) 423). PMID- 15779830 TI - A specific enzyme hydrolyzing 6-O(4-O)-indole-3-ylacetyl-beta-D-glucose in immature kernels of Zea mays. AB - The purification of 6-O(4-O)-indole-3-ylacetyl-beta-D-glucose (IAGlc) hydrolase from immature kernels of maize (Zea mays) was undertaken to separate this enzyme from 1-O-IAGlc hydrolase and beta-glucosidase. Partially purified 6-O(4-O)-IAGlc hydrolase was found to be the specific enzyme catalyzing hydrolysis of stable esters of IAA and glucose. Among a range of ester conjugates tested as substrates, only 6-O(4-O)-IAA-glucose and IBA-glucose isomers were effectively hydrolyzed. No activity against p-nitrophenyl-beta-D-glucopyranoside, a synthetic substrate for beta-glucosidase, was detected in the enzyme preparation. The enzyme is probably involved in the regulation of the IAA levels by the target release of free auxin from ester-linked conjugates, its inactive storage forms. PMID- 15779831 TI - Isolation of trehalose-6-phosphate phosphatase gene from tobacco and its functional analysis in yeast cells. AB - Trehalose is a nonreducing disaccharide composed of two glucose units joined by an alpha, alpha-1, 1 linkage, and has been found in bacteria, yeast, fungi, invertebrates and plants. Accumulation of trehalose in organisms plays a role in enhancing the stress tolerance. A trehalose-6-phosphate phosphatase gene, NtTPPL, was isolated from tobacco in this report. The predicted NtTPPL protein has a putative trehalose_PPase domain. The transcription of the NtTPPL gene was significantly induced by heat stress, and was only slightly induced by NaCl, PEG and low-temperature treatments. When expressing in yeast tps2 mutant, NtTPPL rescued the mutant phenotype under high temperature. This result indicated that NtTPPL functioned as a trehalose-6-phosphate phosphatase in yeast and may play similar roles in plants. PMID- 15779832 TI - Investigation of Nicotiana tabacum (+) N. suaveolens cybrids with carpelloid stamens. AB - To investigate cytoplasmic effects on homeotic floral morphology, Nicotiana tabacum and N. suaveolens protoplasts were fused and cybrids obtained to contrast with the sexual alloplasmic line Nta(sua)S. Nta(sua)S contains the nucleus of N. tabacum and cytoplasm of N. suaveolens while cybrids derive from fused cells where the cytoplasms can interact. The three male-sterile somatic cybrid plants analyzed contained mitochondria with N. tabacum and N. suaveolens mtDNA sequences, but not all the N. tabacum or all the N. suaveolens mtDNA sequences were present. The flowers were N. tabacum-like but with a split corolla (not observed in Nta(sua)S) and the whorl of stamens replaced by a whorl of carpel like structures. Based on scanning electron microscopy the carpelloid stamens had a characteristic N. tabacum stigma, a style of variable length and a pseudo-ovary with ovule-like structures. The Southern blot data were consistent with mtDNA recombination. These genomic changes were maternally inherited. Chloroplasts were either of the N. tabacum or N. suaveolens type. AFLP analysis showed transfer of variable amounts of N. suaveolens nuclear DNA. However, it is the presence of the N. suaveolens sequences and/or absence of N. tabacum sequences in the mitochondria that correlates with the homeotic floral morphology. These cybrids will facilitate the analysis of the role of mitochondrial DNA sequences in floral organ identity; which has received limited attention in genetic flowering models based primarily on Arabidopsis research. PMID- 15779833 TI - Stable maintenance and expression of a foreign gene in transgenic pear shoots retrieved from in vitro conservation. AB - Shoot-tips of transgenic pear were successfully conserved in vitro by slow-growth and cryopreservation methods. After 1 year of storage in slow growth conditions, all shoot-tips survived showing shoot re-growth. Similarly, shoot-tips showed high survival and regeneration rates after cryopreservation. The genetic stability of the transgenic GUS construct in shoots recovered from slow-growth and cryopreservation was assessed. The results from polymerase chain reaction and Southern blotting showed the stable maintenance of the GUS gene in genome, and the Single-Strand Conformation Polymorphism assay did not detect single base variation. X-Gluc staining suggested a normal expression of the GUS gene in shoots retrieved from these storage methods. PMID- 15779834 TI - Correlation of Narcotrend Index, entropy measures, and spectral parameters with calculated propofol effect-site concentrations during induction of propofol remifentanil anaesthesia. AB - OBJECTIVE: The aim of this study was to compare the EEG parameter Narcotrend Index with the spectral and entropy-based EEG parameters median frequency, 95% spectral edge frequency, burst-compensated 95% spectral edge frequency, spectral entropy, amplitude entropy, and approximate entropy with regard to their ability to describe cerebral anaesthetic drug effects during induction of propofol remifentanil anaesthesia. METHODS: Three induction schemes were studied with 10 patients each receiving 2 mg propofol/kg/60s (group 1), 4 mg/kg/120s (group 2), and 4 mg/kg/240s (group 3). The EEG was recorded with the EEG monitor Narcotrend. To analyse the relation between drug effect and EEG parameters, Spearman rank correlation of the different EEG parameters with the calculated propofol effect site concentration was computed. RESULTS: In all groups Narcotrend Index showed the highest correlation with the propofol effect-site concentration and the lowest variability of individual correlation values. Furthermore, only the Narcotrend Index showed a monophasic behaviour over the entire time period analysed. In the group of entropy parameters approximate entropy yielded the best results. Among the spectral parameters the burst-compensated 95% spectral edge frequency had the highest correlation with the propofol effect-site concentration. It was markedly higher than for the standard spectral edge frequency. The correlations of median frequency and amplitude entropy with propofol effect-site concentration were the lowest. CONCLUSIONS. Changes in the propofol effect-site concentration during induction of anaesthesia were best described by the multivariate Narcotrend Index compared to conventional spectral EEG parameters and different entropy measures. PMID- 15779835 TI - Model-based prediction of expiratory resistance index in patients with asthma. AB - OBJECTIVES: Develop a sensitive algorithm and index for detection of asthma patients using forced expiratory flow curves. METHODS: A lumped-parameter model of forced expiration was developed. The model can predict the flow-volume curve during forced expiratory maneuver. The flow-volume curves generated by the model depend on values of resistance parameters (FER). Use of flow-volume curves recorded from normal subjects and from patients with asthma before and after ventolin inhalation as inputs for the inverse model, yielded the resistance parameters for each case. These parameters are based on the entire information presented in the flow-volume curve and on the reduction in flow at all lung volumes. RESULTS: Forced Expiratory Resistance (FER(N)) indices were estimated at different percent of lung volumes using the inverse model. The index was significantly affected by inhalation of ventolin in asthmatic patients and was insensitive to ventolin inhalation in normal patients. In asthmatic patients, the FER index at five lung volumes (out of eight), was two--five times greater than in normal subjects with p < 0.05 (three of them with p < 0.01). CONCLUSIONS: The estimated parameters were sensitive indicators of the degree of lung function impairment and were able to accurately distinguish between healthy and asthmatic patients. PMID- 15779836 TI - Presentation of laboratory and sonoclot variables using principal component analysis: identification of hypo- and hypercoagulation in the HELLP syndrome. AB - The HELLP (Hemolysis, Elevated Liver enzyme, and Low Platelet) syndrome requires close monitoring of rapid changes in hemostasis. A bedside viscoelastic test- Sonoclot--was used together with coagulation, liver and hemolysis laboratory analyses in three parturients with the HELLP syndrome up to 10 days postpartum. Principal component analysis (PCA) was used to reduce the dimensionality of this multivariate problem and to visualize this process in a two-dimensional plot. It was possible to follow changes in these variables over time and to show how they changed in relation to 10 typical healthy parturients with normal laboratory and Sonoclot values as well as 20 simulated patients with hypo- or hypercoagubility. The effects of emergency delivery, correction of low plasma-antithrombin with plasma and antithrombin factor concentrate, plasma exchange and individualized dosages of low molecular weight heparin to counteract postpartum Sonoclot detected hypercoagulation were evaluated. The efficiency of each treatment strategy was visualized in the PCA plot by movement towards an area with normocoagulation. In conclusion, PCA of viscoelastic and laboratory coagulation analysis data facilitated the detection of both hypo- and hypercoagulative events and represents an alternative way to evaluate treatment strategies in patients with complex coagulative disorders, like the HELLP syndrome. PMID- 15779837 TI - Validation of a noninvasive blood pressure monitoring device in normotensive and hypertensive pediatric intensive care patients. AB - OBJECTIVE: To evaluate the performance and to define limitations of a noninvasive blood pressure monitoring device in the critically ill pediatric population. METHOD: Patients were included in the study if they were admitted to the Pediatric Intensive Care Unit, were between the ages of 1 month and 18 years with wrist circumferences of > or =10 cm, and had an indwelling arterial line. Patients were excluded if their systolic blood pressure differed by > or =7.5% between their upper extremities. The measurements were collected simultaneously with those from an arterial line by a computer interfaced with the noninvasive blood pressure monitoring system and the patient's monitor. Heart rates were calculated from the recorded pulse waveforms of the arterial lines. Comparison analyses were performed via bias and precision plots of the blood pressure and heart rate data in addition to calculation of Pearson's correlation coefficients and concordance correlation coefficients. As a nonparametric method of comparison, the proportion of measurements that differed by greater than 10% was calculated. Results. Blood pressures and heart rates of 20 patients between the ages of 12 months and 17 years were monitored by a noninvasive blood pressure monitor for 30 min per patient. This data collection resulted in 2015 data points for each blood pressure and heart rate for comparison of methods. Concordance correlation coefficients were the following: systolic blood pressure, 0.93; diastolic blood pressure, 0.93; mean blood pressure, 0.94; and heart rate, 0.85. CONCLUSIONS: The noninvasive blood pressure monitor is capable of producing an accurate blood pressure measurement every 12-15 heartbeats in addition to providing a pulse waveform and digital display of the heart rate. Our study showed good agreement between the methods in the normotensive and hypertensive critically ill pediatric population with a wrist circumference limitation defined at > or =11 cm. PMID- 15779838 TI - Hipocrates: a robust system for the control of neuromuscular blockade. AB - OBJECTIVE: Development of an automatic system (software package Hipocrates) for the control of neuromuscular blockade by continuous infusion of the non depolarising types of muscle relaxant drugs presently used in anaesthesia, namely atracurium, cisatracurium, vecuronium and rocuronium. METHODS: Hipocrates incorporates control strategies based upon classical, adaptive and robust control, as well as a wide range of noise reduction techniques and on-line adaptation to patient-specific characteristics. Therefore, the system provides strong robustness to inter- and intra-individual variability of the patients responses or unexpected circumstances and adaptation to the individual requirements. RESULTS: The control system is easy to set up and to use in a clinical environment. It consists of a portable PC computer, a Datex AS/3 NMT sensor and a B/Braun compact perfusion pump. In the simulation mode the software package incorporates sophisticated generation of pharmacokinetic/pharmacodynamic models driven by simulated drug administration regimes (bolus, continuous infusion and a combination of both). CONCLUSIONS: Hipocrates is an advanced standalone application for the control of neuromuscular blockade with a friendly graphic interface. It has been extensively validated, and it can be used on patients undergoing surgery as well as for simulation studies. Therefore Hipocrates also provides an excellent environment for education and training purposes. PMID- 15779839 TI - Weaning from long-term mechanical ventilation: a nonpulmonary weaning index. AB - OBJECTIVE: Despite the extensive investigations in the area of weaning, clinicians are still struggling with the question of when to begin the process of weaning. Clinical weaning indices designed to predict the weaning potential are most frequently based on pulmonary factors. However, many physiological, respiratory, and mechanical factors also have impact on weaning, but are often overlooked. We suggest a new "nonpulmonary weaning index" (NPWI), which assesses the influence of factors such as blood albumin and total blood protein on the weaning success. METHODS: We assess the information value of 17 clinical and paraclinical indices in a retrospective study covering 151 patients on a long term (at least 7 days) mechanical ventilation. The most informative of those 17 indices are used in the formulation of NPWI. Its threshold differentiates the successful from the unsuccessful weaning trials. RESULTS: From all 17 indices the most significant are: total blood protein, blood albumin, PaO2, hematocrit, lactate, the ratio PaO2/FiO2, hemoglobine, and RUE. The proposed index uses only two of them: blood albumin and total blood protein. It is easily calculated and can easily be tracked in time. It has high sensitivity and specificity. CONCLUSIONS: The results of this study suggest that in the decision whether to attempt weaning from long-term mechanical ventilation, more attention should be paid to the nonpulmonary factors. PMID- 15779840 TI - The correlation of the bispectral index with propofol effect site concentrations is not altered by epochs indicated as artefact-loaded by narcotrend. AB - OBJECTIVE: Artefact detection is an essential feature of automatic EEG monitoring systems used in anaesthesia. Clinical experience indicates that Narcotrend monitoring (MonitorTechnik, Bad Bramstedt, Germany, version 4.0) excludes more EEG epochs because of artefacts than bispectral index monitoring (BIS, Aspect Medical Systems, Newton, MA, version XP). Whether this increased exclusion of epochs is justified has not been investigated yet. METHODS: Eighteen adult patients undergoing radical prostatectomy were investigated. Induction of anaesthesia was performed with a fentanyl bolus and a propofol infusion. Additionally, following intubation patients received 15 ml bupivacaine 0.5% epidurally. After a waiting period of 45 min depth of anaesthesia was varied two times by increasing and decreasing propofol concentrations. Narcotrend index, BIS values and calculated propofol effect site concentrations were automatically recorded at intervals of 5 s. We tested the hypothesis whether exclusion of artefacts detected by the Narcotrend monitor would possibly improve the prediction probability of the BIS monitor, justifying the necessity of artefact suppression. RESULTS: Simulated propofol effect site concentrations ranged from 2 microg/ml to 6 microg/ml. The Narcotrend monitor excluded a significantly higher percentage of epochs because of artefact detection (12.6 + 1.0%) than the BIS monitor (0.4 +/- 0.1%). The performance of BIS as an indicator of predicted propofol effect site concentrations did not differ when including (P(K) = 0.86 +/ 0.05) or excluding (P(K) = 0.85 +/- 0.04) the data pairs where Narcotrend monitor but not BIS monitor indicated an artefact. Artefacts were evenly distributed over the investigated range ofpropofol effect site concentrations. CONCLUSION: Exclusion of data pairs that were detected as artefacts by Narcotrend but not by BIS did not change the performance of bispectral index as an indicator of propofol effect site concentration. PMID- 15779841 TI - The pulse in reflectance pulse oximetry: modeling and experimental studies. AB - OBJECTIVE: Reflectance pulse oximetry permits the use of alternative monitoring sites such as the face or torso, and is the approach commonly employed in fetal pulse oximetry systems. The purpose of this study is to investigate the impact of assumptions about the nature of arterial pulsatility on the calibration of such systems. METHODS: Monte Carlo simulations of reflectance pulse oximetry were run on a six-layer tissue model, varying depth and magnitude of the arterial pulse. SpO2 readings on and off the femoral artery obtained during desaturation studies in newborn piglets were compared to predictions. Results. Monte Carlo simulation results clarified the difference between deep and shallow pulsatility found with photon diffusion models, agreeing with earlier in vivo observations. Significant overestimation of SpO2 <75% and slight underestimation >75% is expected if a sensor is placed on a highly pulsatile site. The on- and off-artery SpO2 readings recorded during desaturation in the newborn piglet follow the model predictions. CONCLUSIONS: The sensitivity of reflectance pulse oximetry calibration to the depth and magnitude of arterial pulsatility reinforces the observation that monitoring site selection is of importance in optimizing reflectance pulse oximetry performance, particularly fetal pulse oximetry. Sites with palpable pulsatility should be avoided. PMID- 15779842 TI - The effects of isoflurane and propofol on intraoperative neurophysiological monitoring during spinal surgery. AB - OBJECTIVES: To compare the effects of isoflurane and propofol on intraoperative neurophysiological monitoring (IONM) during spinal surgery. METHODS: Thirty-five patients were randomly assigned to receive isoflurane (n = 17) or propofol (n = 18) anesthesia. Somatosensory evoked potentials (SEPs) following posterior tibial nerve stimulation were recorded before induction as baselines. Isoflurane concentrations and propofol infusions were adjusted to obtain four pre-determined BIS ranges: 65-55, 55-45, 45-35 and 35-25. For each range, a stable state was maintained for at least 10 min to perform IONM. The SEP latency P40 and amplitude P40-N50, the onset latency and amplitude of transcranial motor evoked potentials (tcMEPs), and threshold intensity of triggered electromyographic activity (EMG) following pedicle screw stimulation were statistically analyzed. RESULTS: Compared with baseline values, P40 latency increased and P40-N50 amplitude decreased after anesthesia with isoflurane or propofol. Isoflurane caused a dose dependent depression of SEPs, but propofol did not. TcMEPs were recordable and stable in all patients receiving propofol in each BIS range, but only recordable in 10 (58.8%) receiving isoflurane with BIS >55, and 3 (17.8%) with BIS <55. No difference was noted in triggered EMG. CONCLUSIONS: Isoflurane inhibited IONM more than propofol. Propofol is recommended for critical spinal surgery, particularly when motor pathway function is monitored. PMID- 15779843 TI - On the sex ratio of mortality: evolution during the period 1970-1999. AB - In the industrialized world large differences between male and female age specific mortality rates exist, favouring the female sex.These differences vary over time and depend on the type of disease. This study intends to define the problem by determining the male/female (M/F) ratio of mortality for different disease groups and 2 time periods, 1970 and 1999. PMID- 15779844 TI - Comments about possible use of human embryonic stem cell-derived cardiomyocytes to direct autologous adult stem cells into the cardiomyogenic lineage. AB - Several studies have shown that cell-transplantation therapy following myocardial infarction has some efficacy in aiding myocardial repair and subsequent recovery of heart function. Large-scale production of human embryonic stem cell-derived cardiomyocytes can potentially provide an abundant supply of donor cells for myocardial transplantation. There are, however, immunological barriers to their use in human clinical therapy.A novel approach would be to look at utilizing human embryonic stem cell-derived cardiomyocytes to reprogram autologous adult stem cells to express cardiomyogenic function, instead of using these directly for transplantation. This could be achieved through a number of novel techniques. Enucleated cytoplasts generated from human embryonic stem cell-derived cardiomyocytes could be fused with autologous adult stem cells to generate cytoplasmic hybrids or cybrids. Adult stem cells could also be temporarily permeabilized and exposed to cytoplasmic extracts derived from these cardiomyocytes. Alternatively, intact cells or enucleated cytoplasts from human embryonic stem cell-derived cardiomyocytes could be co-cultured with adult stem cells in vitro, to provide the cellular contacts and electrical coupling that might enable some degree of trans-differentiation to take place. This review would therefore examine the potential advantages and disadvantages of such a novel approach, in comparison to other more conventional techniques such as the use of exogenous cytokines/growth factors or the use of genetic modulation. PMID- 15779845 TI - Multisite cardiac pacing. A code proposal revisited. PMID- 15779846 TI - Nitrate-stimulated tilt testing enhances the predictive value of the tilt test on the risk of recurrence in patients with suspected vasovagal syncope. AB - OBJECTIVE: Nitrate-stimulated tilt testing may be used to diagnose vasovagal syncope and to guide therapy. However, to date the predictive value of the test is undetermined. This study analyses the risk of recurrence of syncope in vasovagal patients on the basis of a nitrate-stimulated tilt result and other clinical factors. METHODS AND RESULTS: We used a combined passive and nitrate stimulated tilt protocol. One year after the test a questionnaire, asking about recurrence of syncope and other demographic factors, was sent to 131 patients. Passive tilting was found not to be a useful predictor; recurrence rates after a positive and negative test were 26% and 30%, respectively, and non-significant. With the addition of sublingual nitrates the test became a significant predictor; recurrence rates were 34 (34%) with a positive and 4 (13%) with a negative tilt test (p = 0.031). Univariate analysis of the clinical factors revealed that gender and pre-test symptomatology were significant predictors of recurrence. Multivariate analysis revealed no additional relationships. CONCLUSION: In patients with suspected vasovagal syncope, a positive tilt test with sublingual nitrates together with the clinical factors gender and symptomatology, are predictors for recurrence of syncope during one year of follow-up. Without specific therapy the prognosis of vasovagal syncope is good. PMID- 15779847 TI - Intracardiac basic fibroblast growth factor and transforming growth factor-beta 1 mRNA and their proteins expression level in patients with pressure or volume overload right or left ventricular hypertrophy. AB - OBJECTIVES: The aim of the study was to investigate the pathogenic role of intracardiac basic fibroblast growth factor (bFGF) and transforming growth factor beta-1 (TGF beta-1) mRNA and their protein expression level in patients with ventricular volume or pressure-overload. BACKGROUND: Myocardial hypertrophy is responsiveness of cardiomyocytes and interstitial cells to ventricular stress produced by ventricular preload or/and afterload and a series of growth factors. However, the molecular mechanism by which the changes of bFGF and TGF beta-1 mRNA and their protein expression level in patients with volume or pressure-overload lead to distinct forms of cardiac hypertrophy is unclear. METHODS: 32 patients were divided into two groups: 16 patients with cardiac volume overload leading to left ventricular hypertrophy (VG) and 16 patients with pressure overload leading to right ventricular hypertrophy (PG), respectively, as compared with 5 unexpected deaths of noncardiac disease (CG), which is determinated by echocardiography and/or pathologic examination following operation; intracardiac bFGF and TGF beta-1 mRNA and their protein expression level were determined by immunohistochemistry and in situ hybridization as well as image analysis; cardiomyocytes and fibril collagen of type I and III were examined by haemotoxylin and eosin (HE) staining or sirius-red staining. RESULTS: Cardiocyte transverse diameter and fibril collagen of extracellular matrix, intracardiac bFGF and TGF beta-1 mRNA and their protein expression level in patients with volume or pressure-overload were significantly increased in both VG and PG, as compared with CG. CONCLUSION: This is the first paper to suggest that intracardiac bFGF and TGF beta-1 play a pivotal role in patients with pressure or volume-overload leading to right or left ventricular hypertrophy, composed of cardiomyocyte hypertrophy and extracellular matrix proliferation. PMID- 15779848 TI - Different clinical features, biochemical profiles, echocardiographic and elctrocardiographic findings in older and younger patients with idiopathic dilated cardiomyopathy. AB - OBJECTIVE: To study the clinical features, biochemical profiles, echocardiographic and electrocardiographic findings in the elderly and younger with idiopathic dilated cardiomyopathy (IDCM). METHODS: We measured biochemical profiles and reviewed the clinical features, echocardiographic and electrocardiographic findings in 40 elderly (group A) and 70 younger (group B) patients with IDCM. RESULTS: The aldosterone level in group A was higher than in group B (304.8 +/- 69.1 vs. 213.3 +/- 54.5 pmol/l, P < 0.05). Triiodothyronine (T3) and free T3 in group A were lower than in group B (0.78 +/- 0.21 and 2.87 +/ 0.73 vs. 1.26 +/- 0.33 nmol/l and 3.55 +/- 0.64 pmol/l, all P < 0.05). The incidence of ventricular arrhythmia in group A was lower than in group B (61.3% vs. 92.1 %, P < 0.01). The incidence of hypokalaemia and hypomagnesaemia were higher in group A (51.3% and 27.5%). 28.8% patients in group A were susceptible to digitalis intoxication. Disease duration and mean survival period in group A were longer than in group B (11.0 +/- 4.7 and 6.9 +/- 4.2 vs. 5.2 +/- 2.5 y and 3.4 +/- 2.7 y, all P < 0.05). The main cause of death in group A was congestive heart failure (78.9%) and ventricular arrhythmia (61.9%) in group B. CONCLUSIONS: The prognosis in group A was better than in group B. The patients in group A usually showed low T3 syndrome. Ventricular arrhythmia in group A may be due to heart failure, electrolyte imbalance and sympathetic activation. PMID- 15779849 TI - Acute Chagas' disease: immunohistochemical characteristics of T cell infiltrate and its relationship with T. cruzi parasitic antigens. AB - OBJECTIVE: The present work analysed endomyocardial biopsies of patients with acute Chagas' disease in order to evaluate the frequency and intensity of T. cruzi antigens, CD4+ and CD8+ T cells to determine the characteristics of this recurrent disease in Venezuela. MATERIAL AND METHODS: Twelve endomyocardial biopsies of patients with Chagas' disease, 12 to 51 years old, (7M and 5F) were analysed. T. cruzi antigens and CD4+ (helper) and CD8+ (cytotoxic-suppressor) T cells were detected by the immunoperoxidase technique. The presence and intensity of lymphocytic myocarditis was evaluated according to the degree of myocardial fibre injury caused by inflammatory infiltrate. RESULTS: Myocarditis was present in 100% of the cases. The mean numbers of CD4+ T cell and CD8+ T cell were 11.00 (+/- 10.29); 14.69 (+/- 13.08) and the CD4/CD8 T cell ratio was 0.75. T. cruzi antigens were detected in 58%. There was a good correlation between the numbers of CD4 and CD8 T cells of each case and a lack of correlation with the amount of T. cruzi antigens. CONCLUSION: All patients with acute Chagas' disease show some degree of myocarditis that seems to be directly related to the presence of parasitic antigens. Both CD4 and CD8 T cells participate in this process. We are following these patients to see if patients with severe myocarditis and more parasite antigens in the acute phase will develop chronic heart failure. PMID- 15779850 TI - Echinococcosis of the heart: clinical and echocardiographic features in 12 patients. AB - We describe the clinical and echocardiographic features in 12 patients with hydatid cyst of the heart who were screened between 1985 and 2001. Presenting symptoms have a wide variability but the main symptom was precordial pain. The electrocardiogram was always abnormal and showed T wave inversion in 8 cases, ST depression in 5 cases, incomplete right bundle-branch block in one case and atrial fibrillation in one case. Transthoracic echocardiography and transoesophageal echocardiography constitute the imaging procedure of choice for the diagnosis of cardiac hydatid cyst with a high sensibility, specificity. Computed tomography (CT) was performed in all patients and confirmed the echocardiographic data. MRI was performed in one patient with right atrium hydatid cyst location and confirmed the echocardiographic data. CONCLUSION: In the presence of atypical symptomatology in a patient coming from an endemic area of hydatid cyst, the diagnosis of cardiac echinococcosis is possible.TTE and TEE are the imaging procedure of choice and sufficient for the diagnosis of cardiac hydatid cyst. CT and/or MRI provide more information about the extension of echinococcus disease in other intra- or extrathoracic locations. PMID- 15779851 TI - Effect of Chlamydia pneumoniae infection and hyperlipidaemia on the expression of PPARgamma, P50 and c-Fos in aortic endothelial cells in C57bL/6J mice. AB - OBJECTIVE: To investigate the effect of C. pneumoniae infection and/or hyperlipidaemia on the expression of peroxisome proliferator-activated receptor gamma (PPARgamma), nuclear factor-kappa B (NF-kappaB) and activated protein-1 (AP 1) in aortic endothelial cells in C57BL/6J mice. METHODS AND RESULTS: Forty eight, 8-week-old female C57BL/6J mice were divided into four groups:A, B, C and D (each twelve mice). Group A (as blank control) and B were fed a regular diet. Group C and D were fed an atherogenic diet (consisting of 15% fat, 2.5% cholesterol and 0.5% sodium cholate). Group B and D were infected with C. pneumoniae. Fourteen weeks later, the expression of PPARgamma, P50 (subunit of NF kappaB) and c-Fos (subunit of AP-1) was determined by indirect immunofluorescence in the aortic endothelial cells. Slides of aortic sinus were prepared by cryosection, and stained with Sudan IV for examination of atherosclerotic plaque. The score of atherosclerotic plaque was determined by microscopy. The score of atherosclerotic plaque in group B was not increased, while it was significantly higher in groups C and D (P < 0.01), still the score in group D was higher than in group C (P < 0.01). The expression of PPARgamma, NF-kappaB and AP-1 in endothelial cells in aortic sinus was upregulated in group B, C and D, in comparison with group A (P < 0.05). There was no significant difference among groups B, C and D. CONCLUSION: The expression of PPARgamma, NF-kappaB and AP-1 was upregulated in the endothelial cells in mice infected with C. pneumoniae and/or fed with an atherogenic diet. An atherogenic diet or this diet combined with C. pneumoniae infection accelerated the process of atherosclerosis. The diet infected with C. pneumoniae alone would not accelerate this process. PPARgamma might play an anti-atherosclerotic role in this process. PMID- 15779852 TI - Cardiovascular changes in mucopolysaccharidoses in Taiwan. AB - From 1991 to 2002, we managed 72 patients with mucopolysaccharidoses. We retrospectively reviewed the records of 37 for whom detailed physical findings and cardiovascular evaluation were available. Twenty patients had serial electrocardiographic (ECG) examinations, and none had arrhythmias. All 20 patients had low R wave voltage in V6. One type VI patient had ischaemic changes secondary to severe cardiomyopathy at the terminal stage of her disease. Among the 37 patients in whom echocardiography was performed, cardiovascular abnormalities progressed with age, although most had mild clinical signs and symptoms. The most common changes were abnormalities of the mitral and aortic valves hypertrophy of the interventricular septum, especially in types I, II, and VI. During the follow-up period of up to 5 years, 3 type II patients and I type VI patient expired. In at least 3 of these patients, death was related to cardiopulmonary failure. As MPS patients are surviving longer, complete cardiac evaluation at regular intervals is mandatory and should be part of their routine assessment. PMID- 15779853 TI - Aortic distensibility in coronary artery disease and in aortic stenosis. PMID- 15779854 TI - Secondary cardiac tumour originating from laryngeal carcinoma: case report and review of the literature. AB - Although most malignant tumours metastasize to the heart they rarely produce clinical symptoms. Cardiac metastasis of tumours arising from head and neck and particulary of the larynx are extremely rare. Herein we report an unusual case of successfully treated, advanced laryngeal carcinoma with subsequent late symptomatic metastatic disease to the heart. The patient discussed, developed a right ventricular intracavitary tumour with pseudo-infarct pattern on the ECG. Current litterature regarding incidence, diagnostic techniques, treatment modalities and survival rates of secondary heart tumours are reviewed. PMID- 15779855 TI - Combination of a high bolus dose of tirofiban with half-dose thrombolytics for the treatment of subacute stent thrombosis. AB - Acute stent thrombosis is rare and it is usually related to complications during the procedure. Subacute thrombosis is far more common and is associated with a high incidence of acute myocardial infarction and death. Restoration of flow by thrombolysis, emergency bypass surgery or emergency percutaneous transluminal coronary angioplasty (PTCA) has had only limited success with respect to myocardial salvage. We report the case of a patient who suffered from recurrent subacute stent thrombosis, in whom administration of tirofiban at high-dose bolus in association with a half dose of recombinant tissue plasminogen activator succeeded in restoring normal myocardial flow and stable clinical condition. PMID- 15779856 TI - Anomalous left coronary artery arising from the right sinus of valsalva as a cause of ventricular tachycardia. AB - This paper describes a rare case of a young patient with an anomalous left coronary artery (LCA) originating from the right sinus of Valsalva, passing between the aorta and the pulmonary artery. This rare case was discovered during coronary angiography for the evaluation of the patient, after restoration of symptomatic ventricular tachycardia. PMID- 15779857 TI - Variant angina in a 17-year-old male. AB - Chest pain is a common complaint in adolescents. It is mostly from musculoskeletal origin. Variant angina is a rare cause of chest pain in adolescents. Here we report a 17-year-old male with severe chest pain accompanied by transient ST-segment elevation in leads II, III, avF of the electrocardiogram showing/revealing variant angina associated with acute myocardial ischaemia. Data obtained from laboratory tests, including serial cardiac markers, were normal and subsequent cardiac catheterization revealed a normal coronary anatomy. The patient has been asymptomatic since discharge, and treatment with calcium channel antagonists with nitrate seems to assure a good prognosis. PMID- 15779858 TI - Cardiac metastasis from a stage IIIb cervix carcinoma. AB - We report the case of a 37-year-old woman with a right ventricular mass (detected by echocardiography and PET-CT) associated with cervical cancer. After radiation therapy there was no more evidence for a mass in the right ventricle. We wish to stress that, although the outcome for cervical cancer with cardiac metastases is still very poor, prognosis has improved with advanced imaging technology and radiation therapy. PMID- 15779859 TI - Reversible left ventricular hypertrophy after tako-tsubo-like cardiomyopathy. AB - Tako-tsubo-like cardiomyopathy is a newly-recognized enigmatic disease characterized by transient left ventricular dysfunction of a broad area of the apex with a hyperkinetic area around the cardiac base. There is ST-segment elevation with no coronary stenosis. The exact mechanism for this entity remains unknown. Here, we report a case of tako-tsubo-like cardiomyopathy that showed a marked left ventricular hypertrophy (LVH) when the wall motion returned to normal. LVH was normalized at 10 months. The cause of LVH remains unknown. PMID- 15779860 TI - Ezra Greenspan: a trailblazer in combination chemotherapy. PMID- 15779861 TI - Cyclosporine A, alpha-lnterferon and interleukin-2 following chemotherapy with BCNU, DTIC, cisplatin, and tamoxifen: a phase II study in advanced melanoma. AB - Preclinical data suggest that one method of inducing autoimmunity to tumor is the administration and subsequent withdrawal of cyclosporine A following chemotherapy and that this effect may be enhanced with interferon and interleukin-2. Consequently, we performed a phase II trial in patients with advanced melanoma to explore this approach. Thirty-three patients were treated with BCNU (150 mg/m2 iv every 8 weeks), cisplatin (25 mg/m2 iv days 1-3) every 4 weeks, DTIC (220 mg/m2 iv days 1-3 every 4 weeks) along with tamoxifen (10 mg po BID days 1-4). Cyclosporine A at 3 mg/kg/day in two divided doses was given on days 4-21, alpha interferon 1 million units/m2 subcutaneously every other day on days 4-21 and interleukin-2 1 million units/m2 BID subcutaneously days 21-28 were also given. Of the 33 patients, 3 patients (9%) had complete response and 8 patients (24%) had a partial response for a total response rate of 33% (95% confidence interval 18-52%). Median duration of response was 17 months (range 3+ to 24+ months). Six patients continue to show no signs of tumor progression for 3+, 5+, 10+, 24+, 60+, and 72+ months. Toxicity was generally well tolerated and included myelosuppression and fatigue. This regimen is feasible and generally tolerable and has produced an antitumor response rate comparable with inpatient biochemotherapy regimens. PMID- 15779862 TI - Oxaliplatin for pretreated patients with advanced or metastatic pancreatic cancer: a multicenter phase II study. AB - A phase II study was designed to evaluate the efficacy and safety of oxaliplatin as second-line treatment in patients with locally advanced or metastatic pancreatic cancer. Eighteen patients with advanced pancreatic cancer previously treated with gemcitabine-based chemotherapy, received oxaliplatin 130 mg/m2 i.v. every 21 days. Patients were treated until tumor progression or unacceptable toxicity. No objective response was observed among the 18 treated patients. Three (16.7%) patients had stable disease for > 2 months. A clinical benefit response was observed in five (27.7%) patients. Toxicity was mild. Oxaliplatin as second line treatment for patients with unresectable pancreatic cancer is well tolerated and associated with improvement of tumor-related symptoms despite its failure to induce objective responses. LOHP merits further investigation in combination with other drugs as palliative treatment of pretreated patients with advanced pancreatic cancer. PMID- 15779864 TI - Overexpression of glutathione-S-transferase, MGSTII, confers resistance to busulfan and melphalan. AB - A major obstacle to hematopoietic gene therapy is the lack of appropriate in vivo selection protocols that can raise the presently low numbers of gene-altered stem cells to therapeutically useful levels. Overexpression of glutathione-S transferases (GST), in combination with busulfan treatment, may provide an exploitable selection mechanism for hematopoietic gene therapy strategies. GST provides a major route of detoxification of a variety of xenobiotics, including alkylating agents used for myeloablative chemotherapy. The only known route of clearance of busulfan is by GST-mediated conjugation. Using a fibroblast cell line as a model, we have tested the effects of overexpression of three human GST (GSTA1, GSTP1, and MGSTII) on cell survival under a busulfan or melphalan challenge. In two separate assay formats using chronic exposure to busulfan, MGSTII conferred a reproducible twofold selective advantage. GSTA1 and GSTP1 had no effect on busulfan resistance, and melphalan resistance was not affected by expression of any of the GSTs in these assays. In an acute (24-hour) melphalan exposure assay, MGSTII conferred about a twofold selective advantage. Busulfan was not toxic in this assay. RTPCR analysis of human bone marrow CD34+ cells showed that MGSTII is not highly expressed in this stem/early progenitor population. These data indicate that MGSTII may be a useful selective agent in hematopoietic gene therapy. PMID- 15779863 TI - Phase II study of weekly low-dose paclitaxel for relapsed and refractory non Hodgkin's lymphoma: a Wisconsin Oncology Network Study. AB - This study was performed to determine the clinical activity and safety of weekly low-dose paclitaxel (90 mg/m2) given as a 1-hour infusion in patients with relapsed and refractory non-Hodgkin's lymphoma (NHL). Thirty patients were treated on a phase II protocol conducted at the University of Wisconsin Comprehensive Cancer Center and within the Wisconsin Oncology Network (WON). A cycle of therapy was defined as paclitaxel at 90 mg/m2 weekly for 6 consecutive weeks followed by a 2-week rest period. Cycles were repeated as long as there was no disease progression or unacceptable toxicity. In general, the patients were heavily pretreated with a median of 4 prior therapies (range 2-11), and 73% were refractory to the most recent systemic therapy. The median age was 70 (range 44 97). All NHL histological subtypes were eligible. Of the 30 eligible patients enrolled, 26 were evaluable for response and 28 for toxicity. The overall response rate was 23% (95% confidence interval (CI) 9.0-43.7%). One patient had a complete response, and 5 patients had partial responses. The median response duration was 3.2 months (range 1.4-11.8 months). The median event-free survival was 1.9 months. The major toxicity was neuropathy. Despite the limited marrow reserve in this patient population, myelosuppression was minimal. Paclitaxel given in this dose and schedule has modest activity in previously treated non Hodgkin's lymphoma. The response rate appears similar to other reports using different doses and schedules. Myelosuppression appears less with this schedule than with other schedules. PMID- 15779865 TI - The change of intracellular pH is involved in the cisplatin-resistance of human lung adenocarcinoma A549/DDP cells. AB - We had reported that the intracellular pH (pHi) of human lung adenocarcinoma A549 cells, which is sensitive to cisplatin, was more acidic than that of cisplatin resistant A549/DDP cells. The correlation between the change of the pHi and cisplatin-resistance of A549/DDP cells was further studied by altering pHi in consequence of the change of CO2 concentration of the incubator. The pHi alterations of the cells were monitored by using the fluorescence probe of BCECF AM. The results indicated that the pHi was more alkaline at lower CO2 concentration (2% CO2 in the incubator) and more acidic at higher CO2 concentration (8% CO2 in the incubator) for both A549 and A549/DDP cells compared with those of both A549 and A549/DDP cells cultured at 5% CO2 as the normal condition. Accumulation of bodipy-cisplatin, a fluorescence probe used for drug resistance assays, in A549 cells incubated at 2%, 5%, and 8% CO2 was increased 8.4%, 17.4%, and 23.5% compared to A549/DDP cells, respectively. Intracellular sequestration and distribution of bodipy-cisplatin imaged by laser scanning confocal microscopy indicated that bodipy-cisplatin was more encapsulated in acidic compartments of A549/DDP cells as shown with acridine orange, a dye that specifically labels acidic organelles in the cells. These results can be further confirmed in liposome systems with different pH gradients. It is proposed from the above results that the change of pHi in especially more acidic compartments in A549/DDP cells involves their cisplatin resistance. PMID- 15779866 TI - Prolonged disease-free and treatment-free survival in platinum-resistant ovarian cancer following extended (>1 year) administration of single-agent paclitaxel: A case report and discussion of potential clinical implications. AB - BACKGROUND: The optimal duration of therapy for women with platinum-resistant ovarian cancer who respond to second-line treatment programs remains undefined. CASE REPORT: A patient with well-documented platinum-resistant ovarian cancer who responded to single-agent paclitaxel (175 mg/m2 over 3 hours q 28-35 days) was continued on this program for a total of 15 cycles delivered over a period of 16 months. The patient has subsequently remained without evidence of disease recurrence >7 years following discontinuation of this treatment regimen. CONCLUSION: The prolonged administration of paclitaxel, a highly cycle-specific cytotoxic agent, is a rational management option in patients with platinum resistant ovarian cancer who exhibit evidence of a response to therapy and who do not experience excessive treatment-associated toxicity (e.g., grade 2-3 peripheral neuropathy). PMID- 15779867 TI - Pathology of the future: molecular profiling for targeted therapy. AB - Recent evidence suggests that each patient's cancer has a unique subset of molecular pathogenetic derangements. These derangements may both genetic and proteomic alterations. Genomic and proteomic research tools enable genome-wide assessment of gene expression as well as kinase driven cell signaling events. These tools are illuminating the molecular derangements of individual tumors, even if these tumors have similar morphological characteristics. A combination of laser capture microdissection with multiplexed phosphoproteomic analysis using reverse phase protein microarray technology is being used to identify protein molecular signatures of individual tumors. The in vivo state of multiple kinase driven signal pathways may be evaluated by reverse phase protein microarray with a panel of specific antibodies developed based upon our knowledge of biological processes. Molecular profiling of individual patient's tumors is currently being evaluated in clinical trials at the National Institutes of Health, National Cancer Institute for monitoring Epidermal Growth Factor (EGF) cell signaling events for patients with breast and ovarian cancer. PMID- 15779868 TI - Developing an understanding of proteomics: an introduction to biological mass spectrometry. AB - Three components form the foundation of the rapidly growing field of proteomics research: traditional protein chemistry experiments, separations science, and biological mass spectrometry. The increasing demand for proteomics, as well as the presentation of the 2003 Nobel Prize to John Fenn and Koichi Tanaka for ion source development, has placed biological mass spectrometry in the scientific spotlight. A basic understanding of mass spectrometry is increasingly important for basic, translational, and clinical research. This review article presents an introduction to mass spectrometry, a brief description of different mass spectrometry experiments, and new developments in the field. PMID- 15779869 TI - Environmental and genetic risk factors for childhood leukemia: appraising the evidence. AB - Childhood leukemia is the most common cause of malignancy under the age of 15, representing an annual incidence rate of 43 cases per million in the United States. Confirmed clinical and epidemiologic associations explain less than 10% of disease incidence, leaving 90% of cases with an unclear etiology. To effectively study leukemia in children, one must recognize that this disease has a multifactorial causal mechanism and a heterogeneous biological composition. In addition, the timing of environmental exposures and genetic changes related to disease risk must be considered. This review of both environmental and genetic risk factors for childhood leukemia evaluates the current published literature and synthesizes the available knowledge. Furthermore, attention is directed to expected sources of new advances and the compelling current issues that need to be addressed before further progress can be made. We discuss parental occupational exposures, air pollution, other chemical exposures such as household solvents and pesticides, radiation, dietary factors, immunological factors, socioeconomic status, and genetic susceptibility. We hope to provide the reader with an understanding of the challenge and promise that characterizes the current and future directions in childhood leukemia research. PMID- 15779870 TI - The problem of post-breast cancer lymphedema: impact and measurement issues. AB - Breast cancer treatment is recognized as the most common cause of secondary lymphedema (LE) in the developed countries of the world. LE occurs as both an acute and chronic health condition in which significant and persistent swelling is associated with an abnormal accumulation of protein-rich fluid. The impact of LE is significant on a wide range of daily activities and survivorship quality of life. Measurement and quantification of LE has been problematic despite the fact that various methods have been used to measure limb volume (LV). Perhaps in part because of difficulties in measurement and diagnosis, the reported incidence of LE varies greatly among persons treated with surgery and radiation for breast cancer. Through increased measurement accuracy, LE incidence and prevalence following current therapeutic approaches for breast cancer treatment cancer will be better understood, and more informed decisions about risk factors, treatment interventions, and recovery will be made. Further identification of epidemiological and clinical factors associated with risk and incidence will provide the necessary foundation for preventive intervention. Bilateral measurements at pre-op and over time are necessary to assess LV changes during follow-up, as it is important as part of the differential diagnosis to note whether volume change has occurred in the affected limb alone or in both limbs. Application of rigorous measurement protocols, assessment of symptom experience, and establishment of a data base on bilateral LV at pre-op for later comparison are essential components of a solid foundation for intervention studies. Through multidisciplinary collaboration with rigorous scientific approaches feasible to be carried out in the clinical setting, we have the opportunity to better target risk factors for development of LE, design data-based interventions, and improve post-treatment quality of life. PMID- 15779871 TI - Pain in cancer patients unrelated to the cancer or treatment. AB - The majority of patients with cancer will experience pain in the course of their disease [Kjaer, M. The therapy of cancer pain and its integration into a comprehensive supportive care strategy. Ann. Oncol. 1997, 8 (3), 15-19; Bruera, E.; Lawlor, P. Cancer pain management. Acta Anaesthesiol. Scand. 1997, 41 (1 of 2), 146-153]. Epidemiological studies [Foley, K.M. The treatment of pain in the patient with cancer. CA Cancer J. Clin. 1986, 36 (4), 194-215; Walley, B.A.; Hagen, N.A. The epidemiology of cancer pain. Pain Dig. 1995, (5) 237-244; Portenoy, R.K. Cancer pain: epidemiology and syndromes. Cancer 1989, 63 (11), 2298-2307] generally categorize the pain as 1) directly caused by the neoplastic process or related phenomena; 2) by treatment; or 3) unrelated to the neoplastic process. In approximately 10% of cancer patients who have pain, the pain is unrelated to the disease or treatment and is most often caused by muscles and connective tissue (Twycross, R. Pain Relief in Advanced Cancer; Churchill Livingstone: New York, 1994; 55-61). An overview of pathophysiological mechanisms of muscle pain is presented, followed by a structured protocol to treat frequently encountered pain of muscular origin. The purpose of this article is to provide to the practicing clinicians easy to apply approaches for the treatment of muscle-related pain. PMID- 15779872 TI - Ignorance is bliss? Ethical considerations in therapeutic nondisclosure. AB - It is not uncommon for oncologists and other clinicians to encounter families who wish that grave medical information be withheld from their loved ones. Despite families' typically benevolent motives in making these requests, often, nondisclosure is not in the patients' best interests. Furthermore, nondisclosure may challenge physicians' professional obligations to their patients. However, there are particular situations in which nondisclosure is ethically appropriate, if not ethically obligatory. This article discusses potential benefits and harms associated with therapeutic nondisclosure in adult patients and offers strategies to effect more appropriate disclosure when therapeutic nondisclosure is contemplated. PMID- 15779873 TI - Etiology of childhood leukemia: environment, genes, controversies, and conundrums. PMID- 15779874 TI - Vinca alkaloids vs. taxanes as therapy in lymphoid malignancies: do our experimental models obscure our understanding of how these drugs really work? PMID- 15779875 TI - The relationship between prosodic and syntactic organization in early multiword speech. AB - Several descriptions of the transition from single to multiword utterances use prosody as an important diagnostic criterion. For example, in contrast to successive single-word utterances, 'real' two-word utterances are supposed to be characterized by a unifying intonation contour and a lack of an intervening pause. Research on the acquisition of prosody, however, revealed that control of the phonetic parameters pitch, loudness, and duration is far from complete at such an early stage. In this study, we examine the interaction between the development of different types of syntactic structures and their prosodic organization. Data from a detailed production record of a monolingual German learning boy is analysed both auditorily and acoustically with a focus on four different types of two-word utterances produced between 2;0 and 2;3. Two major findings are reported here. First, the different types of two-word utterances undergo individual trajectories of prosodic (re-)organization, in part depending on the time course in which they become productive. This suggests that different types of multiword utterances become prosodically fluent at different points in time. Second, the variability of prosodic features such as pauses and stress pattern is very high at the onset of combinatorial speech. Consequently, fluency or disfluency of individual examples should not be used as a reliable criterion for their syntactic status and we recommend caution when taking prosody as a cue for syntactic development. PMID- 15779876 TI - Testing the role of semantic similarity in syntactic development. AB - The study explored early syntactic development, and tested the hypothesis that children use similarity of meaning in order to move beyond the learning of individual item-based multiword constructions. The first 6 types of verb-object (VO) constructions in Hebrew-speaking children were analysed for the occurrence of transfer of learning and facilitation, as well as for the semantic similarity of the direct objects (DO). Longitudinal naturalistic speech corpora of 20 children (1;06-2;06) were analysed. We found facilitation (increased rate of learning) among the first 6 types of VO constructions (each type built on a different verb) as evidenced by the accelerating growth curves. Next, we measured the semantic similarity of the DOs using an 8-category system including Patient, Theme and Object of Result. The first 6 DO types represented 3.95 different semantic roles. On the average, after the first VO construction was learned, 3 out of the following 5 constructions produced were not preceded by another VO construction where the DO was of the same semantic category. The results indicate that facilitation of learning of early syntax is most probably NOT mediated by semantic similarity. PMID- 15779877 TI - Maternal responsive and directive behaviours and utterances as predictors of children's lexical development. AB - Predictive relations were examined between measures of 20 mothers' behavioural and verbal general and specific responsiveness and intrusive and supportive directiveness and their children's subsequent expressive vocabularies during three developmental periods with endpoints at the beginning, middle, and end of the second year: 0; 10 to 1; 1, 1 ; 1 to 1; 5, and 1;5 to 1;9. Regression analyses, controlling for mothers' utterance frequencies and children's initial lexicons, revealed considerable consistency between reported and observed lexicons but changing patterns of predictive relations with development. During the first period, behavioural, but not verbal, measures of maternal responsiveness and supportive directiveness were positively predictive. In period two, verbal, but not behavioural, measures predicted children's vocabularies, with specific responsiveness and supportive directiveness as positive predictors and intrusive directiveness as a negative predictor. During the final period, mothers' behavioural and verbal responsiveness and behavioural supportive directiveness positively predicted and their verbal intrusive directiveness negatively predicted children's lexical growth. PMID- 15779878 TI - When children reach beyond their grasp: why some children make pronoun case errors and others don't. AB - This research addresses the question of why some children are disposed to making a large number of pronoun case errors and others are not. The answer proposed is that when pronoun paradigm building outstrips the development of INFL, children become especially vulnerable to erring in the choice of pronominal word form, resulting in pronoun case error. On the other hand, when pronoun paradigm building proceeds more conservatively, the risk of error is reduced. The spontaneous sentence production of children observed in naturalistic caregiver- child interaction from a cross-section of 44 children (2;0-4;0) is used to support this proposal. The data show that pronoun case error was minimal among children who had strong INFL. However, among children with weak INFL there was a wide range of variation, some children making many errors and others making none. Analysis of variance confirmed that this variation was strongly related to the dispersion of production attempts across an extended pronoun paradigm, such that, the fewer cells attempted, the lower the error rate. These findings show that pronoun case errors are not an inevitable result of grammatical development, but may conceivably be avoided altogether if paradigm building proceeds at a rate commensurate with the child's development of INFL. PMID- 15779879 TI - Production of the English past tense by children with language comprehension impairments. AB - Three experiments investigated the ability of eight-year old children with poor language comprehension to produce past tense forms of verbs. Twenty children selected as poor comprehenders were compared to 20 age-matched control children. Although the poor comprehenders performed less well than controls on a range of tasks considered to tap verbal-semantic abilities, the two groups showed equivalent phonological skills. Poor comprehenders performed as well as control children when asked to inflect novel verbs and regular verbs. In contrast, poor comprehenders were less skilled than controls at inflecting both high frequency and low frequency irregular verbs. Although the predominant error pattern for all children was to over-regularize, this was most marked in the poor comprehenders; control children were more likely to produce errors that contained knowledge of the irregular form than poor comprehenders. In addition, the ability to inflect irregular verbs was related to individual differences in verbal-semantic skills. These findings are discussed within a framework in which verb inflection is related to underlying language skills in both the phonological and semantic domains. PMID- 15779880 TI - Large constituent families help children parse compounds. AB - The family size of the constituents of compound words, or the number of compounds sharing the constituents, has been shown to affect adults' access to compound words in the mental lexicon. The present study was designed to see if family size would affect children's segmentation of compounds. Twenty-five English-speaking children between 3;7 and 5;9 were asked to explain the meaning of existing compounds with constituents of varying family size to an alien puppet. The results showed that children were more likely to mention the modifier of compounds if they came from large constituent families than if they came from small constituent families. Other variables were also shown to have some, but smaller effects on children's parsing, including the frequency of the constituent words and the compounds, whether the compounds were already known, and age. These results suggest that children's segmentation of compounds might be facilitated by analogy with other compounds already in their vocabularies. PMID- 15779881 TI - Phonological specificity in children at 1;2. AB - The specificity of infants' phonological representations is examined by comparing their sensitivity to mispronunciations of novel and familiar words, using the preferential looking task. 29 children at 1; 2 were trained and tested on familiar and novel word-object pairs. Children showed evidence of sensitivity to mispronunciations of novel and familiar words, indicating detailed phonological representations. Discrepancies between this study and earlier investigations are discussed with reference to differences between habituation and preferential looking tasks. PMID- 15779882 TI - 'Word-learning wizardry' at 1;6. AB - This article explores whether infants are able to learn words as rapidly as has been reported for preschoolers. Sixty-four infants aged 1;6 were taught labels for either two moving images or two still images. Each image-label pair was presented three times, after which comprehension was assessed using an adaptation of the intermodal preferential looking paradigm. Three repetitions of each label were found to be sufficient for learning to occur, fewer than has previously been reported for infants under two years. Moreover, contrary to a previous finding, learning was equally rapid for infants who were taught labels for moving versus still images. The findings indicate that infants in the early stages of acquiring a vocabulary learn new word-referent associations with ease, and that the learning conditions that allow such learning are less restricted that was previously believed. PMID- 15779883 TI - Productivity and beyond: mastering the Polish genitive inflection. AB - This study charts the development of the genitive masculine inflection, one of the most irregular parts of the Polish case-marking system. 72 Polish children aged from 2;3 to 10;8 participated in a nonce word production experiment testing their ability to supply the genitive form and their sensitivity to the semantic factors determining the choice of ending. Results indicate that productivity, or the ability to supply the inflected form of some nonce words, emerges early: 78% of the two-year-olds were able to inflect at least one test item. However, mastery, or the ability to consistently supply the correct ending, takes considerably longer to develop, and adultlike levels of provision are not reached until about age 10;0. PMID- 15779884 TI - The influence of object pre-exposure on two-year-olds' disambiguation of novel labels. AB - We investigated whether manipulating the perceived novelty of nameless objects would influence two-year-olds' tendency to map novel words to these objects. In Experiment 1, children who had been pre-exposed to target nameless objects were more likely to map novel words onto those objects than children who were not pre exposed to the objects or children who were pre-exposed to non-target members of the nameless object categories. In Experiment 2, children who were pre-exposed to a nameless object were more likely to assign the novel label to that object than to either a familiar object or an unfamiliar object that had not been pre exposed. The results of these studies suggest that reducing the novelty of nameless objects increases two-year-olds' tendency to map a novel word to a nameless object. PMID- 15779885 TI - Object name learning and object perception: a deficit in late talkers. AB - Two experiments examined the relation between early object name learning and the ability to represent objects by their abstract shapes. In Experiment 1, two-year old children with productive vocabularies in the bottom 20th percentile--'late talkers'--were compared with (1) same-age children with larger vocabularies, and (2) younger children matched for productive vocabulary, on their ability to recognize named common objects. Object categories were represented two ways: by lifelike, perceptually rich toys, and by grey caricatures of those objects' abstract shapes. All 3 groups recognized lifelike objects equally well. Both typically-developing control groups were better than late talkers at recognizing shape caricatures of objects whose names they knew. In Experiment 2, late talkers and age-matched controls identified named objects represented by lifelike toys and by duplicates of those toys covered in grey textured paint. Age-matched controls knew more of the object names overall, but both they and the late talkers performed equally well on both kinds of test objects. Thus, late talkers had some difficulty in Experiment 1 recognizing objects from abstract shape cues, but no difficulty in Experiment 2 when the shape cues were realistic. The findings imply a relation between the growth of productive vocabulary and the emergence of the ability to represent object categories by abstract shape. PMID- 15779886 TI - Mapping and binding analysis of peptides derived from the tumor-associated antigen survivin for eight HLA alleles. AB - There is considerable interest in developing immunotherapeutic approaches to elicit tumor-specific CTL responses in cancer patients. Epitope-based approaches aim to deliver the antigenic peptides or epitopes recognized by CTLs rather than the intact tumor antigen. Many tumor-associated proteins are nonmutated self proteins for which the dominant peptide epitopes are usually poorly immunogenic. The subdominant epitopes, however, can elicit robust T cell responses if optimized for their ability to bind to class I MHC molecules. Only the epitopes for a few tumor antigens expressed in human cancers have been defined to this level, mainly for technical reasons. The means to rapidly screen and characterize the binding of epitopes derived from complex tumor-associated antigens is an important enabling technology. Here, we have used the high-throughput technology iTopia to identify those peptides derived from the tumor-associated antigen survivin that bind 8 class I alleles. A library of overlapping nonamers spanning the length of the survivin protein was initially screened for peptides capable of binding each allele. Nineteen HLA-A*0201, zero HLA-A*0101, seven HLA-A*0301, twelve HLA-A*1101, twenty-four HLA-A*2402, six HLA-B*0702, six HLA-B*0801, and eight HLA-B*1501 binding peptides were identified based on an arbitrary cutoff. Peptides capable of binding a given allele were further characterized by their affinity for MHC class I molecules and by the rate of dissociation of the complex. This information should help guide functional studies and future epitope based immunotherapies. PMID- 15779887 TI - Crystal structure of a hyperthermophilic archaeal acylphosphatase from Pyrococcus horikoshii--structural insights into enzymatic catalysis, thermostability, and dimerization. AB - Acylphosphatases catalyze the hydrolysis of the carboxyl-phosphate bond in acyl phosphates. Although acylphosphatase-like sequences are found in all three domains of life, no structure of acylphosphatase has been reported for bacteria and archaea so far. Here, we report the characterization of enzymatic activities and crystal structure of an archaeal acylphosphatase. A putative acylphosphatase gene (PhAcP) was cloned from the genomic DNA of Pyrococcus horikoshii and was expressed in Escherichia coli. Enzymatic parameters of the recombinant PhAcP were measured using benzoyl phosphate as the substrate. Our data suggest that, while PhAcP is less efficient than other mammalian homologues at 25 degrees C, the thermophilic enzyme is fully active at the optimal growth temperature (98 degrees C) of P. horikoshii. PhAcP is extremely stable; its apparent melting temperature was 111.5 degrees C and free energy of unfolding at 25 degrees C was 54 kJ mol( )(1). The 1.5 A crystal structure of PhAcP adopts an alpha/beta sandwich fold that is common to other acylphosphatases. PhAcP forms a dimer in the crystal structure via antiparallel association of strand 4. Structural comparison to mesophilic acylphosphatases reveals significant differences in the conformation of the L5 loop connecting strands 4 and 5. The extreme thermostability of PhAcP can be attributed to an extensive ion-pair network consisting of 13 charge residues on the beta sheet of the protein. The reduced catalytic efficiency of PhAcP at 25 degrees C may be due to a less flexible active-site residue, Arg20, which forms a salt bridge to the C-terminal carboxyl group. New insights into catalysis were gained by docking acetyl phosphate to the active site of PhAcP. PMID- 15779888 TI - Mutant-selective thyromimetics for the chemical rescue of thyroid hormone receptor mutants associated with resistance to thyroid hormone. AB - The thyroid hormone receptors (TRs) are ligand-dependent transcription factors that control the expression of multiple genes involved in development and homeostasis in response to thyroid hormone (triiodothyronine, T3). Mutations to TRbeta that reduce or abolish ligand-dependent transactivation function are associated with resistance to thyroid hormone (RTH), an autosomal dominant human genetic disease. A series of neutral alcohol-based compounds, based on the halogen-free thyromimetic GC-1, have been designed, synthesized, and evaluated in cell-based assays for their ability to selectively rescue three of the most common RTH-associated mutations (i.e., Arg320 --> Cys, Arg320 --> His, and Arg316 --> His) that affect the basic carboxylate-binding arginine cluster of TRbeta. Several analogues show improved potency and activity in the mutant receptors relative to the parent compound GC-1. Most significantly, two of these mutant complementing thyromimics show high potency and activity with a strong preference for the mutant receptors over wild-type TRalpha(wt), that is associated with the cardiotoxic actions of T3. The compounds were evaluated in reporter gene assays using the four common thyroid hormone response elements, DR4, PAL, F2 (LAP), and TSH, and show activities and selectivites consistent with their unique potential as agents to selectively rescue thyroid function to these RTH-associated mutants. PMID- 15779889 TI - Crystal structures of deoxy and CO-bound bjFixLH reveal details of ligand recognition and signaling. AB - Rhizobia directly regulate the expression of genes required for symbiotic nitrogen fixation in response to oxygen concentration via the sensor protein FixL. The N-terminal PAS domain of FixL contains a histidine-coordinated heme and regulates the activity of its effector domain, a C-terminal histidine kinase, in response to binding of oxygen and other ligands at the heme. To further investigate ligand-induced inhibition of FixL, we have determined the crystal structures of the heme domain in both the deoxy state and bound to carbon monoxide, a weak inhibitor of FixL kinase activity. Structures collected at room temperature are presented in each state from two crystallographic space groups at 1.8 and 2 A resolution. These structures reveal displacement of the residues of the H(beta) and I(beta) strands by Leu236 upon CO binding, and this structural change propagates more than 15 A to a region of the structure implicated in signal transduction in PAS proteins. Displacement of residues Ile215, Ile216, and Gly217 in the FG loop is also evident, accompanied by the movement of heme propionate 6 upon change in iron ligation. CO binding increases the temperature factors in the FG loop of the protein and disorders the side chain of Arg206, a conserved residue involved in the FG loop switch mechanism. We relate these results to structural changes in other PAS sensor domains and their involvement in catalytic control. PMID- 15779891 TI - Structural and functional modeling of human lysozyme reveals a unique nonapeptide, HL9, with anti-HIV activity. AB - We previously reported that lysozyme accounts for anti-HIV activity associated with the beta-core fraction of human chorionic gonadotropin [Lee-Huang, S., Huang, P. L., Sun, Y., Kung, H. F., Blithe, D. L. & Chen, H. C. (1999) Proc Natl Acad Sci U S A 96, 2678-81]. To define the structural and sequence requirements for anti-HIV activity, we carried out peptide fragmentation and activity mapping of human lysozyme. We identified two peptides that consist of 18 and 9 amino acids of human lysozyme (HL18 and HL9), corresponding to residues 98-115 and 107 115. HL18 and HL9 are potent inhibitors of HIV-1 infection and replication with EC(50)s of 50 to 55 nM, comparable to intact lysozyme. Scrambling the sequence or substitution of key arginine or tryptophan residues results in loss of antiviral activity. HL9, with the sequence RAWVAWRNR, is the smallest peptide we identified with full anti-HIV activity. It forms a pocket with its basic residues on the surface of the molecule. HL9 exists as an alpha-helix in native human lysozyme, in a region of the protein distinct from the muramidase catalytic site. Monte Carlo peptide folding energy minimizing simulation modeling and CD studies indicate that helical propensity does not correlate with antiviral activity. HL9 blocks HIV-1 viral entrance and replication, and modulates gene expression of HIV infected cells, affecting pathways involved in survival, stress, TGFbeta, p53, NFkappaB, protein kinase C and hedgehog signaling. PMID- 15779890 TI - S-Nitrosation and regulation of inducible nitric oxide synthase. AB - The inducible isoform of nitric oxide synthase (iNOS) and three zinc tetrathiolate mutants (C104A, C109A, and C104A/C109A) were expressed in Escherichia coli and purified. The mutants were found by ICP-AES and the zinc specific PAR colorimetric assay to be zinc free, whereas the wild-type iNOS zinc content was 0.38 +/- 0.01 mol of Zn/mol of iNOS dimer. The cysteine mutants (C104A and C109A) had an activity within error of wild-type iNOS (2.24 +/- 0.12 micromol of NO min(-1) mg(-1)), but the double cysteine mutant had a modestly decreased activity (1.75 +/- 0.14 micromol of NO min(-1) mg(-1)). To determine if NO could stimulate release of zinc and dimer dissociation, wild-type protein was allowed to react with an NO donor, DEA/NO, followed by buffer exchange. ICP-AES of samples treated with 10 microM DEA/NO showed a decrease in zinc content (0.23 +/- 0.01 to 0.09 +/- 0.01 mol of Zn/mol of iNOS dimer) with no loss of heme iron. Gel filtration of wild-type iNOS treated similarly resulted in approximately 20% more monomeric iNOS compared to a DEA-treated sample. Only wild-type iNOS had decreased activity (42 +/- 2%) after reaction with 50 microM DEA/NO compared to a control sample. Using the biotin switch method under the same conditions, only wild-type iNOS had increased levels of S-biotinylation. S-Biotinylation was mapped to C104 and C109 on wild-type iNOS using LysC digestion and MALDI-TOF/TOF MS. Immunoprecipitation of iNOS from the mouse macrophage cell line, RAW-264.7, and the biotin switch method were used to confirm endogenous S-nitrosation of iNOS. The data show that S-nitrosation of the zinc tetrathiolate cysteine results in zinc release from the dimer interface and formation of inactive monomers, suggesting that this mode of inhibition might occur in vivo. PMID- 15779892 TI - Slow proton transfer through the pathways for pumped protons in cytochrome c oxidase induces suicide inactivation of the enzyme. AB - In the absence of subunit III the aa(3)-type cytochrome c oxidase exhibits a shortened catalytic life span (total number of turnovers) due to an increased probability of undergoing irreversible inactivation during steady-state turnover. Inactivation results from structural alteration of the heme a(3)-Cu(B) active site in subunit I [Hosler (2004) Biochim. Biophys. Acta 1655, 332-339]. The absence of subunit III also dramatically slows proton uptake to the active site via the D proton pathway, as well as inhibiting the proton backflow/exit pathway that connects the active site/proton pump with the outer surface of the oxidase complex. Here we demonstrate that these phenomena are linked: slow proton delivery to the active site through these pathways induces suicide inactivation, thus shortening the catalytic life span of the enzyme. Mutations that inhibit the D pathway, but not the K pathway, increase the probability of suicide inactivation. Strong inhibition of the D pathway allows suicide inactivation to occur even in the presence of subunit III. Arachidonic acid, which stimulates proton uptake by the D pathway, retards suicide inactivation. Steady-state turnover in the presence of DeltaPsi and DeltapH, which inhibits proton uptake from the inner surface of the protein, enhances suicide inactivation. Simultaneous inhibition of proton uptake from both sides of the protein by a double mutation affecting the D pathway and the proton backflow/exit pathway greatly shortens the catalytic life span of the oxidase even in the presence of subunit III. Thus, maintenance of rapid proton transfer through the D pathway and the backflow/exit pathway is one mechanism by which subunit III normally functions to prevent suicide inactivation of cytochrome c oxidase. The experiments suggest that increased lifetimes of the heme a(3) oxoferryl intermediates as well as the anionic form of Glu286 of the D pathway cause suicide inactivation in the active site. PMID- 15779893 TI - "Zn-link": a metal-sharing interface that organizes the quaternary structure and catalytic site of the endoribonuclease, RNase E. AB - Ribonuclease E is an essential hydrolytic endonuclease in Escherichia coli, and it plays a central role in maintaining the balance and composition of the messenger RNA population. The enzyme is also required for rRNA and tRNA processing. We have shown earlier that the highly conserved catalytic domain of E. coli RNase E is a homotetramer [Callaghan, A. J. et al. (2003) Biochemistry 42, 13848-13855]. Here, we report that this quaternary organization requires zinc. Two protomers share a single zinc ion, and quantitative analysis indicates that each protein contributes two cysteine thiols toward the coordination of the metal. The candidate cysteines are part of a motif that is conserved in the RNase E protein family, and mutation of these residues causes the partial loss of zinc, the complete disruption of the tetramer into dimers, and effective catalytic inactivation. However, these mutations do not affect RNA binding. The tetramer can be artificially maintained by disulfide bond formation, which fully displaces the zinc but largely preserves the catalytic activity. Thus, catalytic activity does not require zinc directly but does require the quaternary structure, for which the metal is essential. We propose that the RNase E tetramer has two nonequivalent subunit interfaces, one of which is mediated by a single, tetrathiol-zinc complex, which we refer to as a "Zn-link" motif. One or both interfaces organize the active site, which is distinct from the primary site of RNA binding. PMID- 15779894 TI - A tryptophan that modulates tetrahydrobiopterin-dependent electron transfer in nitric oxide synthase regulates enzyme catalysis by additional mechanisms. AB - Nitric oxide synthases (NOSs) are flavo-heme enzymes that require (6R) tetrahydrobiopterin (H(4)B) for activity. Our single-catalytic turnover study with the inducible NOS oxygenase domain showed that a conserved Trp that interacts with H(4)B (Trp457 in mouse inducible NOS) regulates the kinetics of electron transfer between H(4)B and an enzyme heme-dioxy intermediate, and this in turn alters the kinetics and extent of Arg hydroxylation [Wang, Z.-Q., et al. (2001) Biochemistry 40, 12819-12825]. To investigate the impact of these effects on NADPH-driven NO synthesis by NOS, we generated and characterized the W457A mutant of inducible NOS and the corresponding W678A and W678F mutants of neuronal NOS. Mutant defects in protein solubility and dimerization were overcome by purifying them in the presence of sufficient Arg and H(4)B, enabling us to study their physical and catalytic profiles. Optical spectra of the ferric, ferrous, heme-dioxy, ferrous-NO, ferric-NO, and ferrous-CO forms of each mutant were similar to that of the wild type. However, the mutants had higher apparent K(m) values for H(4)B and in one mutant for Arg (W457A). They all had lower NO synthesis activities, uncoupled NADPH consumption, and a slower and less prominent buildup of enzyme heme-NO complex during steady-state catalysis. Further analyses showed the mutants had normal or near-normal heme midpoint potential and heme-NO complex reactivity with O(2), but had somewhat slower ferric heme reduction rates and markedly slower reactivities of their heme-dioxy intermediate. We conclude that the conserved Trp (1) has similar roles in two different NOS isozymes and (2) regulates delivery of both electrons required for O(2) activation (i.e., kinetics of ferric heme reduction by the NOS flavoprotein domain and reduction of the heme-dioxy intermediate by H(4)B). However, its regulation of H(4)B electron transfer is most important because this ensures efficient coupling of NADPH oxidation and NO synthesis by NOS. PMID- 15779895 TI - Thermostability and Ca2+ binding properties of wild type and heterologously expressed PsbO protein from cyanobacterial photosystem II. AB - Oxygenic photosynthesis takes place in the thylakoid membrane of cyanobacteria, algae, and higher plants. Initially light is absorbed by an oligomeric pigment protein complex designated as photosystem II (PSII), which catalyzes light induced water cleavage under release of molecular oxygen for the biosphere on our planet. The membrane-extrinsic manganese stabilizing protein (PsbO) is associated on the lumenal side of the thylakoids close to the redox-active (Mn)(4)Ca cluster at the catalytically active site of PSII. Recombinant PsbO from the thermophilic cyanobacterium Thermosynechococcus elongatus was expressed in Escherichia coli and spectroscopically characterized. The secondary structure of recombinant PsbO (recPsbO) was analyzed in the absence and presence of Ca(2+) using Fourier transform infrared spectroscopy (FTIR) and circular dichroism spectropolarimetry (CD). No significant structural changes could be observed when the PSII subunit was titrated with Ca(2+) in vitro. These findings are compared with data for spinach PsbO. Our results are discussed in the light of the recent 3D-structural analysis of the oxygen-evolving PSII and structural/thermodynamic differences between the two homologous proteins from thermophilic cyanobacteria and plants. PMID- 15779896 TI - Circular permutation and deletion studies of myoglobin indicate that the correct position of its N-terminus is required for native stability and solubility but not for native-like heme binding and folding. AB - We studied the effect of deleted and circularly permuted mutations in sperm whale myoglobin and present here results on three classes of mutants: (i) a deletion mutant, Mb(1)(-)(99), in which the C-terminal helices, G and H, were removed; (ii) two circular permutations, Mb-B_GHA, in which helix B is N-terminal and helix A is C-terminal, and Mb-C_GHAB, in which helix C is N-terminal and helices A and B are C-terminal; and (iii) a deleted circular permutation, Mb-HAB_F, in which helix H is N-terminal, helix F is C-terminal, and helix G is deleted. The conformational characteristics of the apo and holo forms of these mutants were determined at neutral pH, by spectroscopic and hydrodynamic methods. The apo form of the deleted and permuted mutants exhibited a stronger tendency to aggregate and had lower ellipticity than the wild type. The mutants retained the ability to bind heme, but only the circularly permuted holoproteins had native-like heme binding and folding. These results agree with the theory that myoglobin has a central core that is able to bind heme, but also indicate that the presence of N- and C-terminal helices is necessary for native-like heme pocket formation. Because the holopermuteins were less stable than the wild-type protein and aggregated, we propose that the native position of the N-terminus is important for the precise structural architecture of myoglobin. PMID- 15779897 TI - Determining the origin of the stabilization of DNA by 5-aminopropynylation of pyrimidines. AB - DNA duplexes are stabilized by aminopropynyl modification of pyrimidines at the 5 position. A combination of thermodynamic analyses as a function of ionic strength, NMR, and molecular modeling has been applied to determine the origin of the stabilization. UV melting studies of a dodecamer bearing one, two, or three nonadjacent modified dU and dC and of a single dU(8) in the Dickerson-Drew dodecamer revealed that the modifications are essentially additive in terms of T(m), DeltaG, and DeltaH, and there is little difference between dU and dC. The free energy change was parsed into electrostatic and nonelectrostatic components, which showed a significant contribution from charge interactions at physiological ionic strength but also a nonelectrostatic contribution that arises in part from hydration. NMR spectroscopy of the modified Dickerson-Drew dodecamer revealed that the conformation of the duplexes is not significantly altered by the modifications, though (31)P NMR shows that the positive charge may affect ionic interactions with the oxygen atoms of the neighboring phosphates. The modified duplex showed significant hydration in both major and minor grooves. The single strands were also analyzed by NMR, which showed evidence of significant stacking interactions in the modified oligonucleotide. Parsing the energy contribution has shown that electrostatics and hydration can produce substantial increases in thermodynamic stability without significant changes in the conformation of the duplex state. These considerations have significance for the design of oligonucleotides used for hybridization. PMID- 15779898 TI - Correlations among morphology, beta-sheet stability, and molecular structure in prion peptide aggregates. AB - The misfolding of proteins into beta-sheets and the subsequent aggregation of these sheets into fibrous networks underlies many diseases. In this paper, the role of peptide structure in determining the ordering of beta-sheet aggregates and the morphology of fibrils and protofibrils is dissected. Using a series of peptides based on residues 109-122 of the Syrian hamster prion protein (H1) with a range of substitutions at position 117, the link between side chain interactions and beta-sheet thermal stability has been investigated. The thermal stability of beta-sheets is associated with the peptides' ability to adopt the same alignment as wild-type H1, with residue 117 in register across all beta strands [Silva, R. A. G. D., Barber-Armstrong, W., and Decatur, S. M. (2003) J. Am. Chem. Soc. 125, 13674-13675]. These aligned strands are capable of forming long, rigid, and twisted fibrils (as visualized by atomic force microscopy) which are thermostable. Peptides which do not adopt this strand alignment aggregate to form thin, flexible, and smooth protofibrils. The ability to form ordered aggregates, and thus to form twisted fibrils, is modulated by the structure of the side chain of residue 117. PMID- 15779899 TI - Characterization of the protein components of Nephila clavipes dragline silk. AB - Spider silk is predominantly composed of structural proteins called spider fibroins or spidroins. The major ampullate silk that forms the dragline and the cobweb's frame threads of Nephila clavipes is believed to be a composite of two spidroins, designated as Masp 1 and 2. Specific antibodies indeed revealed the presence of Masp 1 and 2 specific epitopes in the spinning dope and solubilized threads. In contrast, sequencing of specific peptides obtained from solubilized threads or gland urea extracts were exclusively homologous to segments of Masp 1, suggesting that this protein is more abundantly expressed in silk than Masp 2. The strength of immunoreactivities corroborated this finding. Polypeptides reactive against both Masp 1 and 2 specific antibodies were found to be expressed in the epithelia of the tail and different gland zones and accumulated in the gland secreted material. Both extracts of gland secretion and solubilized threads showed a ladder of polypeptides in the size range of 260-320 kDa in gel electrophoresis under reducing conditions, whereas gel filtration chromatography yielded molecular masses of the proteins of approximately 300-350 kDa. In the absence of a reducing agent, dimeric forms of the spidroins were observed with estimated molecular masses of 420-480 kDa according to gel electrophoresis and 550-650 kDa as determined by gel filtration chromatography. Depending on the preparation, some silk material readily underwent degradation, and polypeptides down to 20 kDa in size and less were detectable. PMID- 15779900 TI - Structural uncoupling between opposing domains of oxidized calmodulin underlies the enhanced binding affinity and inhibition of the plasma membrane Ca-ATPase. AB - Stabilization of the plasma membrane Ca-ATPase (PMCA) in an inactive conformation upon oxidation of multiple methionines in the calcium regulatory protein calmodulin (CaM) is part of an adaptive cellular response to minimize ATP utilization and the generation of reactive oxygen species (ROS) under conditions of oxidative stress. To differentiate oxidant-induced structural changes that selectively modify the amino-terminal domain of CaM from those that modulate the conformational coupling between the opposing domains, we have engineered a tetracysteine binding motif within helix A in the amino-terminal domain of calmodulin (CaM) that permits the selective and rigid attachment of the conformationally sensitive fluorescent probe 4',5'-bis(1,3,2-dithioarsolan-2 yl)fluorescein-(1,2-ethanedithiol)(2) (FlAsH-EDT(2)). The position of the FlAsH label in the amino-terminal domain provides a signal for monitoring its binding to the CaM-binding sequence of the PMCA. Following methionine oxidation, there is an enhanced binding affinity between the amino-terminal domain and the CaM binding sequence of the PMCA. To identify oxidant-induced structural changes, we used frequency domain fluorescence anisotropy measurements to assess the structural coupling between helix A and the amino- and carboxyl-terminal domains of CaM. Helix A undergoes large amplitude motions in apo-CaM; following calcium activation, helix A is immobilized as part of a conformational switch that couples the opposing domains of CaM to stabilize the high-affinity binding cleft associated with target protein binding. Methionine oxidation disrupts the structural coupling between opposing globular domains of CaM, without affecting the calcium-dependent immobilization of helix A associated with activation of the amino-terminal domain to promote high-affinity binding to target proteins. We suggest that this selective disruption of the structural linkage between the opposing globular domains of CaM relieves steric constraints associated with high affinity target binding, permitting the formation of new contact interactions between the amino-terminal domain and the CaM-binding sequence that stabilizes the PMCA in an inhibited conformation. PMID- 15779901 TI - Unusual four-bond secondary H/D isotope effect supports a short-strong hydrogen bond between phospholipase A2 and a transition state analogue inhibitor. AB - A prominent secondary four-bond hydrogen/deuterium isotope effect was observed from proton NMR at the active site histidine imidazole ring of bovine pancreatic sPLA(2) in the presence of a phosphonate transition state analogue. The cross modulation of H(epsilon2)/H48 and H(delta1)/H48 resonances was confirmed by line shape simulation that follows the McConnell equation with fractionation factors incorporated to account for the change in the signal magnitude as well as the resonance line shape at various H(2)O/D(2)O solvent mixtures. While the downfield shift of each individual proton upon deuteration on the opposite site can be attributed to the proton-relay system of the H48-D99 catalytic dyad in sPLA(2), the observation that H(delta1)/H48 induces a 3-fold larger H/D secondary isotope effect ( approximately 0.15 ppm) on H(epsilon2)/H48 than vice versa ( approximately 0.05 ppm) is interpreted as additional spectroscopic evidence for the previously proposed short-strong hydrogen bond formed between the donor N(delta1)/H48 and a nonbridging phosphonate oxygen atom of the transition state analogue. These results provide additional details for the catalytic mechanism of sPLA(2) and demonstrate that the intrinsic H/D secondary isotope effect is a useful tool to probe hydrogen bond strength. PMID- 15779902 TI - Thermochromatium tepidum photoactive yellow protein/bacteriophytochrome/diguanylate cyclase: characterization of the PYP domain. AB - The purple phototrophic bacterium, Thermochromatium tepidum, contains a gene for a chimeric photoactive yellow protein/bacteriophytochrome/diguanylate cyclase (Ppd). We produced the Tc. tepidum PYP domain (Tt PYP) in Escherichia coli, and found that it has a wavelength maximum at 358 nm due to a Leu46 substitution of the color-tuning Glu46 found in the prototypic Halorhodospira halophila PYP (Hh PYP). However, the 358 nm dark-adapted state is in a pH-dependent equilibrium with a yellow species absorbing at 465 nm (pK(a) = 10.2). Following illumination at 358 nm, photocycle kinetics are characterized at pH 7.0 by a small bleach and red shift to what appears to be a long-lived cis intermediate (comparable to the I(2) intermediate in Hh PYP). The recovery to the dark-adapted state has a lifetime of approximately 4 min, which is approximately 1500 times slower than that for Hh PYP. However, when the Tt PYP is illuminated at pH values above 7.5, the light-induced difference spectrum indicates a pH-dependent equilibrium between the I(2) intermediate and a red-shifted 440 nm intermediate. This equilibrium could be responsible for the sigmoidal pH dependence of the recovery of the dark-adapted state (pK(a) = 8.8). In addition, the light-induced difference spectrum shows that, at pH values above 9.3, there is an apparent bleach near 490 nm superimposed on the 358 and 440 nm changes, which we ascribe to the equilibrium between the protonated and ionized dark-adapted forms. The L46E mutant of Tt PYP has a wavelength maximum at 446 nm, resembling wild-type Hh PYP. The kinetics of recovery of L46E following illumination with white light are slow (lifetime of 15 min at pH 7), but are comparable to those of wild-type Tt PYP. We conclude that Tt PYP is unique among the PYPs studied to date in that it has a photocycle initiated from a dark-adapted state with a protonated chromophore at physiological pH. However, it is kinetically most similar to Rhodocista centenaria PYP (Ppr) despite the very different absorption spectra due to the lack of E46. PMID- 15779903 TI - Site-directed mutagenesis of active site residues of phosphite dehydrogenase. AB - Phosphite dehydrogenase (PTDH) catalyzes the unusual oxidation of phosphite to phosphate with the concomitant reduction of NAD(+) to NADH. PTDH shares significant amino acid sequence similarity with D-hydroxy acid dehydrogenases (DHs), including strongly conserved catalytic residues His292, Glu266, and Arg237. Site-directed mutagenesis studies corroborate the essential role of His292 as all mutants of this residue were completely inactive. Histidine selective inactivation studies with diethyl pyrocarbonate provide further evidence regarding the importance of His292. This residue is most likely the active site base that deprotonates the water nucleophile. Kinetic analysis of mutants in which Arg237 was changed to Leu, Lys, His, and Gln revealed that Arg237 is involved in substrate binding. These results agree with the typical role of this residue in D-hydroxy acid DHs. However, Glu266 does not play the typical role of increasing the pK(a) of His292 to enhance substrate binding and catalysis as the Glu266Gln mutant displayed an increased k(cat) and unchanged pH rate profile compared to those of wild-type PTDH. The role of Glu266 is likely the positioning of His292 and Arg237 with which it forms hydrogen bonds in a homology model. Homology modeling suggests that Lys76 may also be involved in substrate binding, and this postulate is supported by mutagenesis studies. All mutants of Lys76 display reduced activity with large effects on the K(m) for phosphite, and Lys76Cys could be chemically rescued by alkylation with 2 bromoethylamine. Whereas a positively charged residue is absolutely essential for activity at the position of Arg237, Lys76 mutants that lacked a positively charged side chain still had activity, indicating that it is less important for binding and catalysis. These results highlight the versatility of nature's catalytic scaffolds, as a common framework with modest changes allows PTDH to catalyze its unusual nucleophilic displacement reaction and d-hydroxy acid DHs to oxidize alcohols to ketones. PMID- 15779905 TI - Characterization of three essential residues in the conserved ATP-binding region of Epstein-Barr virus thymidine kinase. AB - The thymidine kinase encoded by Epstein-Barr virus (EBV TK) is an important target for antiviral therapy and the treatment of EBV-associated malignancies. Through computer-assisted alignment with other human herpesviral TK proteins, EBV TK was shown to contain a conserved ATP-binding motif as for the other TK enzymes. To investigate functional roles of three highly conserved residues (G294, K297, T298) within this region, site-directed mutagenesis was employed to generate various mutants. The TK enzyme activity and ATP-binding ability of these mutant TK enzymes were determined and compared with EBV wild-type TK (wtTK). Mutant G294V lost its ATP-binding ability and was inactive in enzyme activity assay. As the enzyme activity of G294A was reduced to 20% of that of wtTK, the K(m) for ATP binding of G294A was 48.7 microM as compared with 30.0 microM of EBV wtTK. These results suggested that G294 participates in ATP binding and contributes to maintenance of structure. EBV TK mutants K297E, K297Q, and K297R lost their ATP-binding ability and enzyme activity. However, K297R was shown to have a preference for usage of GTP (K(m): 43.0 microM) instead of ATP (K(m): 87.6 microM) as the phosphate donor. This implies that, in addition to nucleotide binding, K297 was involved in the selection of phosphate donor. While EBV TK mutant T298S retained approximately 80% of wtTK enzyme activity, T298A lost its enzyme activity, suggesting that a hydroxyl group at this position is important for the enzyme activity. Interestingly, T298A retained its ATP-binding ability, suggesting a role of T298 in the catalytic process but not in the coordination of ATP. This study demonstrated that amino acid residues G294, K297, and T298 in the ATP-binding motif of EBV TK enzyme are essential for the enzymatic activity but are involved in different aspects of its action. PMID- 15779904 TI - Role of putative anion-binding sites in cytoplasmic and extracellular channels of Natronomonas pharaonis halorhodopsin. AB - Natronomonas (Natronobacterium) pharaonis halorhodopsin (NpHR) is an inward light driven Cl(-) ion pump. For efficient Cl(-) transport, the existence of Cl(-) binding or -interacting sites in both extracellular (EC) and cytoplasmic (CP) channels is postulated. Candidates include Arg123 and Thr126 in EC channels and Lys215 and Thr218 in CP channels. The roles played by these amino acid residues in anion binding and in the photocycle have been investigated by mutation of the amino acid residues at these positions. Anion binding was assayed by changes in circular dichroism and the shift in the absorption maximum upon addition of Cl(-) to anion-free NpHR. The binding affinity was affected in mutants in which certain EC residues had been replaced; this finding revealed the importance of Arg123. On the other hand, mutants in which certain residues in the CP channel were replaced (CP mutants) did not show changes in their dissociation constants. The photocycles of these mutants were also examined, and in the case of the EC mutants, the transition to the last step was greatly delayed; on the other hand, in the CP mutants, L2-photointermediate decay was significantly prolonged, except in the case of K215Q, which lacked the O-photointermediate. The importance of Thr218 for binding of Cl(-) to the CP channel was indicated by these results. On the basis of these observations, the possible anion transport mechanism of NpHR was discussed. PMID- 15779906 TI - Effect of mutation of the Sac7d intercalating residues on the temperature dependence of DNA distortion and binding thermodynamics. AB - Sac7d is a small chromatin protein from the hyperthermophile Sulfolobus acidocaldarius which kinks duplex DNA by approximately 66 degrees at a single base pair step with intercalation of V26 and M29 side chains. Site-directed mutagenesis coupled with calorimetric and spectroscopic data has been used to characterize the influence of the intercalating side chains on the structure and thermodynamics of the DNA complex from 5 to 85 degrees C. Two single-alanine substitutions (V26A and M29A) and five double-glycine, -alanine, -leucine, phenylalanine, and -tryptophan substitutions of the surface residues have been created. NMR and fluorescence titrations indicated that the substitutions had little effect on the structure of the protein or DNA binding site size. Each of the mutant proteins demonstrated a temperature-dependent binding enthalpy which was correlated with a similar temperature dependence in the structure of the complex reflected by changes in fluorescence and circular dichroism. A positive heat capacity change (DeltaC(p)) for DNA binding was observed for only those mutants which also demonstrated a thermotropic structural transition in the complex, and the temperature range for the positive DeltaC(p) coincided with that observed for the structural transition. The thermodynamic data are interpreted using a model in which binding is linked to an endothermic distortion of the DNA in the complex. The results support the proposal that the unfavorable enthalpy of binding of Sac7d at 25 degrees C is due in part to the distortion of DNA. PMID- 15779907 TI - Toward the full set of human mitochondrial aminoacyl-tRNA synthetases: characterization of AspRS and TyrRS. AB - The human mitochondrion possesses a translational machinery devoted to the synthesis of 13 proteins. While the required tRNAs and rRNAs are produced by transcription of the mitochondrial genome, all other factors needed for protein synthesis are synthesized in the cytosol and imported. This is the case for aminoacyl-tRNA synthetases, the enzymes which esterify their cognate tRNA with the specific amino acid. The genes for the full set of cytosolic aaRSs are well defined, but only nine genes for mitochondrial synthetases are known. Here we describe the genes for human mitochondrial aspartyl- and tyrosyl-tRNA synthetases and the initial characterization of the enzymes. Both belong to the expected class of synthetases, have a dimeric organization, and aminoacylate Escherichia coli tRNAs as well as in vitro transcribed human mitochondrial tRNAs. Genes for the remaining missing synthetases were also found with the exception of glutaminyl-tRNA synthetase. Their sequence analysis confirms and further extends the view that, except for lysyl- and glycyl-tRNA synthetases, human mitochondrial and cytosolic enzymes are coded by two different sets of genes. PMID- 15779908 TI - Structural aspects of RecA-dependent homologous strand exchange involving human telomeric DNA. AB - Telomeric DNA sequences in human cells and those of other vertebrates consist of long d(TTAGGG) repeats. In somatic cells, telomeres shorten every cell division with shortening serving as a mitotic clock that counts cell divisions and ultimately results in cellular senescence. Telomere length is principally maintained by a ribonucleoprotein, telomerase. However, a non-negligible proportion of human cells use a recombination-based mechanism for telomere maintenance, termed alternative maintenance of telomeres (ALT). Although the molecular mechanism of ALT is not known, GT-rich sequences in prokaryotes and eukaryotes display high levels of recombination relative to those of non-GT-rich DNA. We show that human telomeric strand-exchange complexes mediated by Escherichia coli RecA protein differ from those formed with nontelomeric sequences. Moreover, telomeric strand-exchange intermediates, unlike those involving nontelomeric sequences, exhibit a tendency to form higher-order nucleoprotein structures. We propose that the strong DNA unwinding activity inherent in the assembly of the RecA strand-exchange complex promotes the formation of alternative DNA structures at human telomeric loci. Organization of these noncanonical structures into higher-order complexes involving multiple DNA duplexes could facilitate the search for homology on different DNA molecules and provide a framework for understanding recombination-dependent mechanisms of telomere maintenance. PMID- 15779909 TI - Multiple loop conformations of peptides predicted by molecular dynamics simulations are compatible with nuclear magnetic resonance. AB - The affinity and selectivity of protein-protein interactions can be fine-tuned by varying the size, flexibility, and amino acid composition of involved surface loops. As a model for such surface loops, we study the conformational landscape of an octapeptide, whose flexibility is chemically steered by a covalent ring closure integrating an azobenzene dye into and by a disulfide bridge additionally constraining the peptide backbone. Because the covalently integrated azobenzene dyes can be switched by light between a bent cis state and an elongated trans state, six cyclic peptide models of strongly different flexibilities are obtained. The conformational states of these peptide models are sampled by NMR and by unconstrained molecular dynamics (MD) simulations. Prototypical conformations and the free-energy landscapes in the high-dimensional space spanned by the phi/psi angles at the peptide backbone are obtained by clustering techniques from the MD trajectories. Multiple open-loop conformations are shown to be predicted by MD particularly in the very flexible cases and are shown to comply with the NMR data despite the fact that such open-loop conformations are missing in the refined NMR structures. PMID- 15779910 TI - Effect of pH on the active site of an Arg121Cys mutant of the metallo-beta lactamase from Bacillus cereus: implications for the enzyme mechanism. AB - The zinc-dependent metallo-beta-lactamases are a group of bacterial enzymes that pose a threat to the future efficacy of present-day antibiotics. Their mechanism is poorly understood, and there are no clinically useful inhibitors. While most members of the group contain two tightly bound zinc ions in their active sites, the Bacillus cereus enzyme has a much lower affinity for its second zinc (Zn2), thought to be due to the presence of Arg121 immediately beneath the floor of the active site (cf. Cys/Ser/His121 in the bizinc enzymes). Crystal structures of the Arg121Cys mutant of the B. cereus 569/H/9 enzyme were solved at pH 7.0, 5.0, and 4.5, each in the presence of either 20 microM or 20 mM Zn(2+) to generate the mono- and bizinc forms, respectively. Surprisingly, the structure of the active site was unaffected by the mutation; a network of ordered water molecules replaced the interactions made by the arginine side chain, and the occupancy of Zn2 appeared minimally changed. As the pH was lowered, Zn2 moved away from one of its ligands, Asp120, but was "tracked" by two others, Cys221 and His263. Furthermore, the hydroxide ion (and proposed nucleophile for beta-lactam hydrolysis) was bound to Zn1 at pH 5 and above but absent at pH 4.5. This provides experimental evidence for an earlier proposed mechanism in which protonation of Asp120 and the Zn1-bound hydroxide are the two events that lead to the loss of activity at low pH. PMID- 15779911 TI - Evidence for Watson-Crick and not Hoogsteen or wobble base pairing in the selection of nucleotides for insertion opposite pyrimidines and a thymine dimer by yeast DNA pol eta. AB - We have recently reported that pyrene nucleotide is preferentially inserted opposite an abasic site, the 3'-T of a thymine dimer, and most undamaged bases by yeast DNA polymerase eta (pol eta). Because pyrene is a nonpolar molecule with no H-bonding ability, the unusually high efficiencies of dPMP insertion are ascribed to its superior base stacking ability, and underscore the importance of base stacking in the selection of nucleotides by pol eta. To investigate the role of H bonding and base pair geometry in the selection of nucleotides by pol eta, we determined the insertion efficiencies of the base-modified nucleotides 2,6 diaminopurine, 2-aminopurine, 6-chloropurine, and inosine which would make a different number of H-bonds with the template base depending on base pair geometry. Watson-Crick base pairing appears to play an important role in the selection of nucleotide analogues for insertion opposite C and T as evidenced by the decrease in the relative insertion efficiencies with a decrease in the number of Watson-Crick H-bonds and an increase in the number of donor-donor and acceptor acceptor interactions. The selectivity of nucleotide insertion is greater opposite the 5'-T than the 3'-T of the thymine dimer, in accord with previous work suggesting that the 5'-T is held more rigidly than the 3'-T. Furthermore, insertion of A opposite both Ts of the dimer appears to be mediated by Watson Crick base pairing and not by Hoogsteen base pairing based on the almost identical insertion efficiencies of A and 7-deaza-A, the latter of which lacks H bonding capability at N7. The relative efficiencies for insertion of nucleotides that can form Watson-Crick base pairs parallel those for the Klenow fragment, whereas the Klenow fragment more strongly discriminates against mismatches, in accord with its greater shape selectivity. These results underscore the importance of H-bonding and Watson-Crick base pair geometry in the selection of nucleotides by both pol eta and the Klenow fragment, and the lesser role of shape selection in insertion by pol eta due to its more open and less constrained active site. PMID- 15779912 TI - Structural details of the thermophilic filamentous bacteriophage PH75 determined by polarized Raman microspectroscopy. AB - The filamentous virus PH75, which infects the thermophile Thermus thermophilus, consists of a closed DNA strand of 6500 nucleotides encapsidated by 2700 copies of a 46-residue coat subunit (pVIII). The PH75 virion is similar in composition to filamentous viruses infecting mesophilic bacteria but is distinguished by in vivo assembly at 70 degrees C and thermostability to at least 90 degrees C. Structural details of the PH75 assembly are not known, although a fiber X-ray diffraction based model suggests that capsid subunits are highly alpha-helical and organized with the same symmetry (class II) as in the mesophilic filamentous phages Pf1 and Pf3 [Pederson et al. (2001) J. Mol. Biol. 309, 401-421]. This is distinct from the symmetry (class I) of phages fd and M13. We have employed polarized Raman microspectroscopy to obtain further details of PH75 architecture. The spectra are interpreted in combination with known Raman tensors for modes of the pVIII main chain (amide I) and Trp and Tyr side chains to reveal the following structural features of PH75: (i) The average pVIII peptide group is oriented with greater displacement from the virion axis than peptide groups of fd, Pf1, or Pf3. The data correspond to an average helix tilt angle of 25 degrees in PH75 vs 16 degrees in fd, Pf1, and Pf3. (ii) The indolyl ring of Trp 37 in PH75 projects nearly equatorially from the subunit alpha-helix axis, in contrast to the more axial orientations for Trp 26 of fd and Trp 38 of Pf3. (iii) The phenolic rings of Tyr 15 and Tyr 39 project along the subunit helix axis, and one phenoxyl engages in hydrogen-bonding interaction that has no counterpart in either fd or Pf1 tyrosines. Also, in contrast to fd, Pf1, and Pf3, the packaged DNA genome of PH75 exhibits no Raman anisotropy, suggesting that DNA bases are not oriented unidirectionally within the nucleocapsid assembly. The structural findings are discussed in relation to intrasubunit and intersubunit interactions that may confer hyperthermostability to the PH75 virion. A refined molecular model is proposed for the PH75 capsid subunit. PMID- 15779914 TI - Can a hydroxide ligand trigger a change in the coordination number of magnesium ions in biological systems? AB - Density functional (B3LYP) calculations indicate that a hydroxide ligand is capable of triggering a reduction in the coordination number of Mg(2+) ions from 6 to 5. Since this could be quite relevant in the mode of action of magnesium containing enzymes (especially hydrolases in which a metal-bound hydroxide species is believed to play a crucial role), we have performed a systematic deprotonation study of biologically relevant magnesium complexes. We explicitly calculated the preferred coordination number of [MgL(1)(x)L(2)(y)L(3)(z)](2)( )(n) species at the B3LYP/aug-cc-pVTZ level of theory. L(1), L(2), and L(3) represent combinations of water, hydroxide, carboxylate (models Glu and Asp), ammonia ligands (models Lys and His residues), and fluoride ions. As expected, Mg(2+) exclusively prefers an octahedral coordination geometry with H(2)O, HCO(2)(-), or NH(3). Surprisingly, one hydroxide ligand triggers a change to a trigonal bipyramidal geometry. The isoelectronic fluoride ion behaves similarly. When two OH(-) are present, a tetrahedral coordination geometry is preferred. We postulate that a hydroxide (in addition to its role as an active nucleophile) could be employed by magnesium-containing enzymes to trigger a differential coordination behavior. PMID- 15779913 TI - The tertiary structure of the hairpin ribozyme is formed through a slow conformational search. AB - The hairpin ribozyme is a small catalytic RNA comprised of two internal loops carried on two adjacent arms of a four-way helical junction (4WJ). To achieve catalytic activity, the ribozyme folds into a compact conformation that facilitates the formation of tertiary interactions between the two loops. We have investigated the folding kinetics of the natural 4WJ form of the hairpin ribozyme, as well as a minimal construct consisting of just the two loop containing duplexes, by means of stopped-flow fluorescence resonance energy transfer between donor and acceptor probes attached to the ends of the loop bearing arms. Folding was initiated by the addition of Mg(2+) ions or a pseudosubstrate strand to the ribozyme, and the ensuing changes in the emission of both donor and acceptor were monitored over time. Both ribozyme constructs exhibited slow, biphasic kinetic behavior, attributed to two parallel folding pathways leading to compact, docked structures. Two distinct folding rates were observed across a range of Mg(2+) concentrations, and increasing amounts of Mg(2+) accelerated both rates. Notably, both rates were essentially independent of temperature, indicating that the corresponding activation enthalpies were negligible, in contrast to the large activation enthalpies generally observed for RNA folding processes. Instead, the slow folding was due to unfavorable entropy changes in reaching the transition state, indicating that the ribozyme tertiary structure forms through a slow conformational search. These features were observed in both forms of the ribozyme, indicating that the conformational search is confined to the two loop regions and is largely independent of the overall ribozyme architecture. Conformational search may be a general mechanism of tertiary structure formation in RNA. PMID- 15779915 TI - Kinetic analysis and ligand-induced conformational changes in dimeric and tetrameric forms of human thymidine kinase 2. AB - Recombinant human thymidine kinase 2 (hTK2) expressed in Escherichia coli has been found to bind tightly a substoichiometric amount of deoxyribonucleoside triphosphates (dTTP > dCTP >> dATP), known to be strong feedback inhibitors of the enzyme. Incubation of hTK2 with the substrate dThd was able to release the dNTPs from the active site during purification from E. coli and thus allowed the kinetic characterization of the noninhibited enzyme, with the tetrameric hTK2 showing slightly higher activity than the most abundant dimeric form. The unliganded hTK2 revealed a lower structural stability than the inhibitor-bound enzyme forms, being more prone to aggregation, thermal denaturation, and limited proteolysis. Moreover, intrinsic tryptophan fluorescence (ITF), far-UV circular dichroism (CD), and limited proteolysis have revealed that hTK2 undergoes distinct conformational changes upon binding different substrates and inhibitors, which are known to occur in the nucleoside monophosphate kinase family. The CD monitored thermal denaturation of hTK2 dimer/tetramer revealed an irreversible process that can be satisfactorily described by the two-state irreversible denaturation model. On the basis of this model, the parameters of the Arrhenius equation were calculated, providing evidence for a significant structural stabilization of the enzyme upon ligand binding (dCyd < MgdCTP < dThd < dCTP < dTTP < MgdTTP), whereas MgATP further destabilizes the enzyme. Finally, surface plasmon resonance (SPR) was used to study in real time the reversible binding of substrates and inhibitors to the immobilized enzyme. The binding affinities for the inhibitors were found to be 1-2 orders of magnitude higher than for the corresponding substrates, both by SPR and ITF analysis. PMID- 15779916 TI - Role of the second intradiscal loop of peripherin/rds in homo and hetero associations. AB - Peripherin/rds (P/rds) is a disk rim protein that assembles into homo and hetero complexes with its nonglycosylated homologue, Rom-1, to maintain the integrity of the photoreceptor outer segment. Mutations in the rds gene have been identified in a variety of human retinal degenerative diseases. More than 70% of these mutations are located in the second intradiscal (D2) loop, highlighting the functional importance of this region. This study examines the involvement of different regions of the D2 loop in protein associations using a GST pull-down assay and a heterologous coexpression system. The pull-down assay suggests an association of the N-terminal portion (Phe(120)-Phe(187)) of the D2 loop with Rom 1 as well as with other P/rds molecules. Through peptide competition experiments, the region between Cys(165) and Asn(182) of the D2 loop has been identified as the domain for these associations. In a COS-1 cell heterologous expression system, coexpression of the D2 loop along with the intact P/rds and Rom-1 hindered the association of the two full-length proteins. In contrast to the homo association of P/rds molecules, it seems that the hetero association of P/rds with Rom-1 has a more stringent structural requirement. This work defines the crucial domain of the D2 loop, which mediates homo and hetero associations, specifically the regions that lay between Cys(165) and Asn(182). Elucidation of the molecular mechanisms behind the protein-protein associations of P/rds and its partners may reveal the pathogenic defects arising from the most common mutations in this gene. PMID- 15779917 TI - Structural requirements of tropomodulin for tropomyosin binding and actin filament capping. AB - Regulation of actin filament dynamics underlies many cellular functions. Tropomodulin together with tropomyosin can cap the pointed, slowly polymerizing, filament end, inhibiting addition or loss of actin monomers. Tropomodulin has an unstructured N-terminal region that binds tropomyosin and a folded C-terminal domain with six leucine-rich repeats. Of tropomodulin 1's 359 amino acids, an N terminal fragment (Tmod1(1)(-)(92)) suffices for in vitro function, even though the C-terminal domain can weakly cap filaments independent of tropomyosin. Except for one short alpha-helix with coiled coil propensity (residues 24-35), the Tmod1(1)(-)(92) solution structure shows that the fragment is disordered and highly flexible. On the basis of the solution structure and predicted secondary structure, we have introduced a series of mutations to determine the structural requirements for tropomyosin binding (using native gels and CD) and filament capping (by measuring actin polymerization using pyrene fluorescence). Tmod1(1)( )(92) fragments with mutations of an interface hydrophobic residue, L27G and L27E, designed to destroy the alpha-helix or coiled coil propensity, lost binding ability to tropomyosin but retained partial capping function in the presence of tropomyosin. Replacement of a flexible region with alpha-helical residues (residues 59-61 mutated to Ala) had no effect on tropomyosin binding but inhibited the capping function. A mutation in a region predicted to be an amphipathic helix (residues 65-75), L71D, destroyed the capping function. The results suggest that molecular flexibility and binding to actin via an amphipathic helix are both required for tropomyosin-dependent capping of the pointed end of the actin filament. PMID- 15779918 TI - Domain formation in sphingomyelin/cholesterol mixed membranes studied by spin label electron spin resonance spectroscopy. AB - Interactions of palmitoylsphingomyelin with cholesterol in multilamellar vesicles have been studied over a wide range of compositions and temperatures in excess water by using electron spin resonance (ESR) spectroscopy. Spin labels bearing the nitroxide free radical group on the 5 or 14 C-atom in either the sn-2 stearoyl chain of phosphatidylcholine (predominantly 1-palmitoyl) or the N stearoyl chain of sphingomyelin were used to determine the mobility and ordering of the lipids in the different phases. Two-component ESR spectra of the 14 position spin labels demonstrate the coexistence first of gel (L(beta)) and liquid-ordered (L(o)) phases and then of liquid-ordered and liquid-disordered (L(alpha)) phases, with progressively increasing temperature. These phase coexistences are detected over a limited range of cholesterol contents. ESR spectra of the 5-position spin labels register an abrupt increase in ordering at the L(alpha)-L(o) transition and a biphasic response at the L(beta)-L(o) transition. Differences in outer splitting between the C14-labeled sphingomyelin and phosphatidylcholine probes are attributed to partial interdigitation of the sphingomyelin N-acyl chains across the bilayer plane in the L(o) state. In the region where the two fluid phases, L(alpha) and L(o), coexist, the rate at which lipids exchange between phases (<<7 x 10(7) s(-)(1)) is much slower than translational rates in the L(alpha) phase, which facilitates resolution of two component spectra. PMID- 15779919 TI - Role of arginine in the stabilization of proteins against aggregation. AB - The amino acid arginine is frequently used as a solution additive to stabilize proteins against aggregation, especially in the process of protein refolding. Despite arginine's prevalence, the mechanism by which it stabilizes proteins is not presently understood. We propose that arginine deters aggregation by slowing protein-protein association reactions, with only a small concomitant effect on protein folding. The associated rate effect was observed experimentally in association of globular proteins (insulin and a monoclonal anti-insulin) and in refolding of carbonic anhydrase. We suggest that this effect arises because arginine is preferentially excluded from protein-protein encounter complexes but not from dissociated protein molecules. Such an effect is predicted by our gap effect theory [Baynes and Trout (2004) Biophys. J. 87, 1631] for "neutral crowder" additives such as arginine which are significantly larger than water but have only a small effect on the free energies of isolated protein molecules. The effect of arginine on refolding of carbonic anhydrase was also shown to be consistent with this hypothesis. PMID- 15779920 TI - Thioesterase portability and peptidyl carrier protein swapping in yersiniabactin synthetase from Yersinia pestis. AB - Multimodular enzymes, including polyketide synthases (PKSs), nonribosomal peptide synthetases (NRPSs), and mixed PKS/NRPS systems, contain functional domains with similar functions. Domain swapping and module fusion are potential powerful strategies for creating hybrid enzymes to synthesize modified natural products. To explore these strategies, yersiniabactin (Ybt) synthetase containing two subunits, HMWP2 [two NRPS modules (N-terminus-ArCP-Cy1-A-PCP1 and Cy2-PCP2-C terminus)] and HMWP1 [one PKS (N-terminus-KS-AT-MT1-KR-ACP) one NRPS module (Cy3 MT2-PCP3-TE-C-terminus)], was used as a model system to study peptidyl carrier protein (PCP) domain swapping, thioesterase (TE) portability, and module-module fusion. The PCP1 domain of the N-terminal NRPS module of HMWP2 was swapped with either PCP2 or PCP3. The fusion proteins were 3-8-fold less active than the wild type protein. The swapping of PCP2 of HMWP2 abolished the heterocyclization activity of the Cy2 domain while retaining its condensation function. When the two PCPs of HMWP2 were swapped by PCP3TE, it created two active fusion proteins: one or two NRPS modules fused to the TE domain. The internal TE domain of the two fusion proteins catalyzed the hydrolysis of enzyme-bound intermediates HPT-S-PCP3 to form HPT-COOH and HPTT-S-PCP3 to form HPTT-COOH. The TE activity was eliminated by the S2980A point mutation at its active site. Therefore, the three PCPs of the Ybt synthetase were swappable, and its lone TE domain was portable. The reasons for the observed low activities of the fusion proteins and lessons for protein engineering in generating novel modular enzymes were discussed. PMID- 15779921 TI - Neuropeptide processing profile in mice lacking prohormone convertase-1. AB - Prohormone convertase 1 (PC1; also known as PC3) is believed to be responsible for the processing of many neuropeptide precursors. To look at the role PC1 plays in neuropeptide processing in brain and pituitary, we used radioimmunoassays (RIA) as well as quantitative peptidomic methods and examined changes in the levels of multiple neuropeptide products in PC1 knockout (KO) mice. The processing of proenkephalin was impaired in PC1 KO mouse brains with a decrease in the level of Met-Enkephalin immunoreactivity (ir-Met-Enk) and an accumulation of higher molecular weight processing intermediates containing ir-Met-Enk. Processing of the neuropeptide precursor VGF was also affected in PC1 KO mouse brains with a decrease in the level of an endogenous 3 kDa C-terminal peptide. In contrast, the processing of proSAAS into PEN was not altered in PC1 KO mouse brains. Quantitative mass spectrometry was used to analyze a number of peptides derived from proopiomelanocortin (POMC), provasopressin, prooxytocin, chromogranin A, chromogranin B, and secretogranin II. Among them, the levels of oxytocin and peptides derived from chromogranin A and B dramatically decreased in the PC1 KO mouse pituitaries, while the levels of peptides derived from proopiomelanocortin and provasopressin did not show substantial changes. In conclusion, these results support the notion that PC1 plays a key role in the processing of multiple neuroendocrine peptide precursors and also reveal the presence of a redundant system in the processing of a number of physiologically important bioactive peptides. PMID- 15779922 TI - Characterization of the aminocoumarin ligase SimL from the simocyclinone pathway and tandem incubation with NovM,P,N from the novobiocin pathway. AB - Simocyclinone D(8) consists of an anguicycline C-glycoside tethered by a tetraene diester linker to an aminocoumarin. Unlike the antibiotics novobiocin, clorobiocin, and coumermycin A(1), the phenolic hydroxyl group of the aminocoumarin in simocyclinone is not glycosylated with a decorated noviosyl moiety that is the pharmacophore for targeting bacterial DNA gyrase. We have expressed the Streptomyces antibioticus simocyclinone ligase SimL, purified it from Escherichia coli, and established its ATP-dependent amide bond forming activity with a variety of polyenoic acids including retinoic acid and fumagillin. We have then used the last three enzymes from the novobiocin pathway, NovM, NovP, and NovN, to convert a SimL product to a novel novobiocin analogue, in which the 3-prenyl-4-hydroxybenzoate of novobiocin is replaced with a tetraenoate moiety, to evaluate antibacterial activity. PMID- 15779924 TI - Reactive surface micropatterning by wet stamping. AB - Hydrogel stamps are used to reactively micropattern various types of substrates. The method, called reactive wet stamping (r-WETS), is general in nature and overcomes several limitations of conventional soft-lithographic techniques. Illustrative applications of r-WETS in surface wettability modification, deposition of metallic microstructures, preparation of supports for electrostatic self-assembly, and multistep reactive patterning are discussed. PMID- 15779923 TI - Improved method for the preparation of carboxylic acid and amine terminated self assembled monolayers of alkanethiolates. AB - We present an improved method to prepare carboxylic acid (COOH) and amine (NH2) terminated self-assembled monolayers (SAMs) of alkanethiolates. In this method, a small amount of CF3COOH (for COOH-SAM) or N(CH2CH3)3 (for NH2-SAM) is added into the ethanolic solution of alkanethiols during SAM formation. The freshly formed COOH- and NH2-SAMs are then rinsed with an ethanolic solution of NH4OH or CH3COOH, respectively. Both SAMs prepared with the improved method show better quality in terms of surface chemical composition, roughness, and wettability as measured by X-ray photoelectron spectroscopy, atomic force microscopy, and contact angle, respectively. The formation of better SAMs can be attributed to the disruption of interplane hydrogen bonds. PMID- 15779925 TI - Self-assembled structure of nanoparticles at a liquid-liquid interface. AB - Pickering emulsions are used as a template to investigate the multiphase interactions and self-assembled structure of nanoparticles at a trichloroethylene water interface. The dodecanethiol-capped silver nanoparticles of 1-5 nm form randomly distributed multilayers at the liquid/liquid interface, with an interparticle distance varying from close contact to approximately 25 nm. This report offers the first direct observation of nanoparticles in a liquid medium using the environmental transmission electron microscope, as well as the first work revealing the detailed self-assembled structure of nanoparticles at a liquid/liquid interface when the size of the nanoparticles is comparable to the molecular dimension of the liquids. PMID- 15779926 TI - Photoinduced one-electron reduction of MV2+ in titania nanosheets using porphyrin in mesoporous silica thin films. AB - Composite films of a meso-(tetramethylpyridinium)porphyrin (TMPyP) hybrid incorporated in mesoporous silica (MPS) and cast on a methyl viologen (MV2+)/titania nanosheet hybrid were synthesized and a light-induced charge separation between the two could be observed. These composite thin films were able to initiate a one-electron reduction of the MV2+ ions accompanied by the simultaneous decomposition of the TMPyP organic dye within the mesoporous silica channels. PMID- 15779927 TI - Formation of graphitic structures in cobalt- and nickel-doped carbon aerogels. AB - We have prepared carbon aerogels (CAs) doped with cobalt or nickel through sol gel polymerization of formaldehyde with the potassium salt of 2,4 dihydroxybenzoic acid, followed by ion exchange with M(NO3)2 (where M = Co2+ or Ni2+), supercritical drying with liquid CO2, and carbonization at temperatures between 400 and 1050 degrees C under a N2 atmosphere. The nanostructures of these metal-doped carbon aerogels were characterized by elemental analysis, nitrogen adsorption, high-resolution transmission electron microscopy (HRTEM), X-ray photoelectron spectroscopy (XPS), and X-ray diffraction (XRD). Metallic nickel and cobalt nanoparticles are generated during the carbonization process at about 400 and 450 degrees C, respectively, forming nanoparticles that are approximately 4 nm in diameter. The sizes and size dispersion of the metal particles increase with increasing carbonization temperatures for both materials. The carbon frameworks of the Ni- and Co-doped aerogels carbonized below 600 degrees C mainly consist of interconnected carbon particles with a size of 15-30 nm. When the samples are pyrolyzed at 1050 degrees C, the growth of graphitic nanoribbons with different curvatures is observed in the Ni- and Co-doped carbon aerogel materials. The distance of graphite layers in the nanoribbons is approximately 0.38 nm. These metal-doped CAs retain the overall open cell structure of metal free CAs, exhibiting high surface areas and pore diameters in the micro- and mesoporic region. PMID- 15779928 TI - Core-shell nanostructures from single poly(N-vinylcaprolactam) macromolecules: stabilization and visualization. AB - Formation of core-shell poly(N-vinylcaprolactam) (PVCL) single-molecule nanostructures due to interaction of PVCL with metal ions was studied using transmission electron microscopy, 13C NMR, and light scattering. This study demonstrates that addition of CoCl2 to PVCL in its globular conformation yields unimolecular core-shell polymer particles with the core decorated with Co(II) ions. The crucial condition for formation of well-defined unimolecular nanostructures is the presence of stable globular aggregates in aqueous solution. Moreover, the metal ions should have a sufficiently high coordination number (higher than 2) to provide a cross-linking and stabilization of the core. PMID- 15779929 TI - Temperature-controlled vesicle aggregation in the mixed system of sodium n dodecyl sulfate/n-dodecyltributylammonium bromide. AB - Temperature-controlled vesicle aggregation was investigated in a catanionic surfactant system of sodium n-dodecyl sulfate/n-dodecyltributylammonium bromide. Vesicle aggregation took place as the temperature reached the critical value (Tc). Tc can be adjusted by the variations of the total surfactant concentration and the mixed molar ratio. It was also found that the temperature variation above Tc can greatly influence the vesicle aggregation rate. The vesicle aggregation process was irreversible as long as T >/= Tc, whereas the vesicle disaggregation process occurred only below Tc. PMID- 15779930 TI - Reactions of C=C-terminated self-assembled monolayers with gas-phase ozone. AB - Reactions of gas-phase ozone with alkene-terminated alkanethiol self-assembled monolayers on Au are explored using reflection-absorption infrared spectroscopy (RAIRS). The experiments are performed by using a directional doser to control the ozone flux onto the surface and in situ high vacuum techniques to eliminate reactions with atmospheric contaminants. We find that reactions between ozone and the C=C terminal group proceed through the formation of a carboxylic acid moiety that subsequently converts to an interchain carboxylic acid anhydride. PMID- 15779931 TI - Probing alpha-helical and beta-sheet structures of peptides at solid/liquid interfaces with SFG. AB - We demonstrated that sum frequency generation (SFG) vibrational spectroscopy can distinguish different secondary structures of proteins or peptides adsorbed at solid/liquid interfaces. The SFG spectrum for tachyplesin I at the polystyrene (PS)/solution interface has a fingerprint peak corresponding to the B1/B3 mode of the antiparallel beta-sheet. This peak disappeared upon the addition of dithiothreitol, which can disrupt the beta-sheet structure. The SFG spectrum indicative of the MSI594 alpha-helical structure was observed at the PS/MSI594 solution interface. This research validates SFG as a powerful technique for revealing detailed secondary structures of interfacial proteins and peptides. PMID- 15779932 TI - Fabrication of superhydrophobic surfaces from microstructured ZnO-based surfaces via a wet-chemical route. AB - The fabrication of a superhydrophobic surface is demonstrated via a wet chemical route, and this method offers advantages of being cleanroom free, cost efficiency, and wide applicability. The preferable growth of ZnO crystalline forms a microstructured surface, and a variety of alkanoic acids were adopted to tune the surface wettability. Although all surfaces show an advancing contact angle greater than 150 degrees , they substantially differ in the wetting mechanisms. It is found that only when the length of alkanoic acid is greater than 16, the microstructured surface shows a stable superhydrophobicity, in which the Cassie state dominates. While for those moderate-length alkanoic acids (C8 C14), their corresponding surfaces have a tendency to fall into the Wenzel state and display a great contact angle hysteresis. PMID- 15779933 TI - Stabilization of lysozyme-incorporated polyion complex micelles by the omega-end derivatization of poly(ethylene glycol)-poly(alpha,beta-aspartic acid) block copolymers with hydrophobic groups. AB - To improve the stability of lysozyme-incorporated polyion complex (PIC) micelles in physiological condition, three types of hydrophobic groups, including phenyl (Phe), naphthyl (Nap), and pyrenyl (Py) terminal groups, were separately introduced to the omega-end of poly(ethylene glycol)-poly(alpha,beta-aspartic acid) block copolymers (PEG-P(Asp)). The goal was to enhance association forces between the enzyme, lysozyme, and PEG-P(Asp) carriers. Introduction of these hydrophobic groups significantly decreases micellar critical association concentration and increases the micellar tolerability against increasing NaCl concentrations. Particularly, PIC micelles formed from PEG-P(Asp) with Py groups was most stable against increasing NaCl concentrations up to 0.1 M. Significant deviation from a spherical shape for the micelles was also observed for the PEG P(Asp)-Py system, consistent with an increased association number. PMID- 15779934 TI - Dediazoniation in SDS/BuOH/H2O reverse micelles: structural parameters, kinetics, and mechanism of the reaction. AB - Dediazoniation of o-methylbenzenediazonium tetrafluoroborate was investigated in SDS/BuOH/H2O (SDS = sodium dodecyl sulfate) reverse micelles, RMs, and, for comparison, in binary BuOH/H2O mixtures by employing a combination of spectrophotometric and chromatographic techniques. RMs were characterized by steady-state fluorescence; the data indicate that the aggregation number of the RMs increase upon increasing [SDS], while the radius of the water pool is mainly controlled by the amount of water in the system, and that the thickness of the interfacial region increases upon increasing the amount of BuOH in the system, in agreement with literature reports. Experimental evidence suggests that dediazoniation mainly takes place in the interfacial region of the RMs. Kinetic data show that a turnover from the heterolytic to the homolytic mechanism takes place about pH = 5; the variation of the observed rate constants, k(obs,) with pH following an S-shaped curve. At pH approximately 2, k(obs) values are insensitive to solvent composition both in RMs and in the binary mixture; however, k(obs) values in RMs are slightly lower than those in BuOH/H2O, probably due to the presence of SDS. High-performance liquid chromatography analyses of the reaction mixture indicate, in both RMs and in binary mixtures, the main dediazoniation products are the heterolytic ArOH and ArOBu, their yields depending on the composition of the system, and only small (<10%) amounts of the reduction ArH product were detected. The data at low pH are interpreted in terms of a DN + AN dediazoniation mechanism, i.e., a rate-limiting formation of an extremely reactive aryl cation that further reacts with available nucleophiles in the solvation shell. PMID- 15779935 TI - Zwitterionic heterogemini surfactants containing ammonium and carboxylate headgroups. 1. Adsorption and micellization. AB - Zwitterionic heterogemini surfactants with two hydrocarbon chains and two different hydrophilic groups, N,N-dimethyl-N-[2-(N'-alkyl-N'-beta carboxypropanoylamino)ethyl]-1-alkylammonium bromides (2C(n)AmCa, where n represents the hydrocarbon chain lengths of 8, 10, 12, and 14), were synthesized by N,N-dimethylethylenediamine with alkyl bromide, followed by reaction with succinic anhydride. One of the hydrophilic groups is a carboxylate anion, and the other is an ammonium cation. Their physicochemical properties were characterized by measuring equilibrium and dynamic surface tension, fluorescence intensity of pyrene, and light-scattering intensity. A relationship between a logarithm of critical micelle concentration (cmc) and hydrocarbon chain length showed a linear decrease upon increasing chain length and then a departure from linearity at n = 14. This is due to the existence of premicellar aggregations at concentrations below the cmc for n = 14. The surface tension of 2C(n)AmCa reached 27-30 mN m(-1) at each cmc, indicating efficiencies typical of hydrocarbon chain surfactants. The adsorbing rate at the air/water interface became slow with an increase of the chain length. From the fluorescence intensity ratios of 373 and 384 nm using pyrene as a probe, for n = 8, 10, and 14, the pyrene was solubilized in surfactant micelles at around the cmc, whereas for n = 12 the pyrene was solubilized from a concentration of 10-fold the cmc. The scattering intensities by dynamic light scattering also increased from around these concentrations for each chain length, showing the formation of aggregates in solution. PMID- 15779937 TI - Influence of the hydrotrope structure on the physical chemical properties of polyoxide aqueous solutions. AB - The physical chemical properties of block substituted poly(ethylene oxide propylene oxide) (PEO-PPO) block copolymer aqueous solutions were evaluated in the presence of two hydrotropes of different structures: sodium p-toluene sulfonate (NaPTS) and butyl monoglycol sodium sulfonate (NaBMGS). The critical micelle concentration and the cloud point of the copolymer solutions were displaced to higher concentration values, indicating that the solubility of the copolymer was increased in the presence of the hydrotropes. Temperature increased the micelle hydrodynamic radius, but concentration had a limited effect. Carbon 13 nuclear magnetic resonance (13C NMR) permitted the interaction between the surface-active agent and the hydrotrope to be evaluated: NaBMGS, which presented a more pronounced hydrotropic effect, interacts more effectively with the hydrophobic moiety of the surfactant, while NaPTS interacts rather mainly with the hydrophilic oxyethylenic groups. The results furnish experimental evidence to conclude that the hydrotropic phenomenon is specific in relation to both the hydrotrope and the solubilizate. PMID- 15779936 TI - Is the ether group hydrophilic or hydrophobic? AB - A series of six surfactants, each with two ether oxygens within otherwise all hydrocarbon chains, were synthesized and examined for their colloidal properties. Since an ether oxygen is sterically and conformationally similar to the methylene group it has replaced, the ether effect on micellization should stem mainly from solvation of the oxygen and, possibly, disrupted hydrophobicity of its adjacent carbons. It was found that critical aggregation values among the surfactants differ only modestly despite the total length of the ether-separated carbon segments ranging from 12 to 18. Shorter ether surfactants with only 12 or 14 total carbons appear to form small, loose aggregates owing, presumably, to a mild hydrophilicity of the ether groups. A surfactant with 18 chain carbons has a greater tendency to associate hydrophobically, but this is counterbalanced by a relatively water-free environment encountered by the ether groups within a more conventional micelle interior. The result is a leveling effect in which the critical aggregation concentration (cac) loses it sensitivity to chain length. Above their cac's, none of the ether surfactants is a good solubilizer of tetramethysilane or mesitylene. This is not necessarily a predictable finding since it was conceivable that the presence of interior ether groups might actually enhance solubilization (much as ether is a better solvent than hexane). Foamability and solid adsorption studies also indicate that the ethers impair surface activity. In response to the question posed in the paper's title, two ether groups are not sufficiently hydrophilic to prevent aggregation, but they do manage to alter the micelles' morphology and properties considerably. PMID- 15779938 TI - Synthesis and phase characterization of a double-tailed pyrrole-containing surfactant: a novel tecton for the production of functional nanostructured materials. AB - A double-tailed polymerizable (pyrrolylalkyl) ammonium amphiphile has been synthesized, and its interfacial properties and aqueous phase behavior have been studied by polarized optical microscopy and X-ray diffraction. The Krafft temperature is about 27 degrees C, and the critical micelle concentration at 40 degrees C is about 1 mM, as obtained from surface tension measurements, potentiometry, and isothermal titration calorimetry. The lyotropic behavior of the surfactant is found to be of a complex nature. At concentrations higher than the micellar (L1) region, two mesophases have been identified: a second isotropic (L2) phase, which is probably micellar but not fully miscible with water, and a lamellar (L(alpha)) phase, showing interesting alignment properties. Small-angle X-ray scattering analysis of the mesophases has been evaluated in terms of a model of spherical micelles, which describes a mutual arrangement by a structure factor derived from a hard-sphere potential (Percus-Yevick, "PY", approach). Interest in the comprehensive phase behavior of the polymerizable surfactant is based on the desire to integrate the system into a composite material to obtain potentially conducting self-assembled hybrid mesostructures. PMID- 15779939 TI - Thermodynamic properties of the unique self-assembly of {Mo72Fe30} inorganic macro-ions in salt-free and salt-containing aqueous solutions. AB - Static and dynamic laser light scattering techniques are used to monitor the slow self-assembly of 2.5-nm-diameter, hollow spherical, fully hydrophilic heteropolyoxometalate {Mo72Fe30} macro-ions into single-layer vesicle-like "blackberries" (averaging approximately 50-60 nm in diameter) in dilute salt-free and salt-containing aqueous solutions, to obtain the thermodynamic properties of the unique self-assembly. A very high activation energy is observed during the transition from the single ion (general solute state) to blackberries (so-called "second solute state"), which might be responsible for the interestingly slow self-assembly process in dilute solutions. The thermodynamic parameters of the blackberry formation can be affected by adding simple electrolytes into the solution, because the electrostatic interactions are responsible for the unique self-assembly, and the effects of various anions and cations (in the low salt concentration regimes) are discussed. Multivalent anions make the single {Mo72Fe30} macro-ions more stable and make the blackberry formation more difficult. Small cations carrying more charges tend to accelerate the self assembly process. This is the first study on the thermodynamic properties of the novel self-assembly in dilute solutions and the equilibrium and transition between the two solute states of macro-ions in solution. PMID- 15779941 TI - High-Q ultrasonic determination of the critical nanoaggregate concentration of asphaltenes and the critical micelle concentration of standard surfactants. AB - Asphaltenes are known to be interfacially active in many circumstances such as at toluene-water interfaces. Furthermore, the term micelle has been used to describe the primary aggregation of asphaltenes in good solvents such as toluene. Nevertheless, there has been significant uncertainty regarding the critical micelle concentration (CMC) of asphaltenes and even whether the micelle concept is appropriate for asphaltenes. To avoid semantic debates we introduce the terminology critical nanoaggregate concentration (CNAC) for asphaltenes. In this report, we investigate asphaltenes and standard surfactants using high-Q, ultrasonic spectroscopy in both aqueous and organic solvents. As expected, standard surfactants are shown to exhibit a sharp break in sonic velocity versus concentration at known CMCs. To prove our methods, we measured known surfactants with CMCs in the range from 0.010 g/L to 2.3 g/L in agreement with the literature. Using density determinations, we obtain micelle compressibilities consistent with previous literature reports. Asphaltenes are also shown to exhibit behavior similar to that of ultrasonic velocity versus concentration as standard surfactants; asphaltene CNACs in toluene occur at roughly 0.1 g/L, although the exact concentration depends on the specific (crude oil) asphaltene. Furthermore, using asphaltene solution densities, we show that asphaltene nanoaggregate compressibilities are similar to micellar compressibilities obtained with standard nonionic surfactants in toluene. These results strongly support the contention that asphaltenes in toluene can be treated roughly within the micelle framework, although asphaltenes may exhibit small levels of aggregation (dimers, etc.) below their CNAC. Furthermore, our extensive results on known surfactants agree with the literature while the asphaltene CNACs reported here are one to two orders of magnitude lower than most previously published results. (Previous work utilized the terminology "micelle" and "CMC" for asphaltenes.) We believe that the previously reported high concentrations for asphaltene CMCs do not correspond to primary aggregation; perhaps they refer to higher levels of aggregation or perhaps to a particular surface structure. PMID- 15779940 TI - Photooxidation of diglycine in confined media. Application of the microreactor model for spin-correlated radical pairs in reverse micelles and water-in-oil microemulsions. AB - Time-resolved electron paramagnetic resonance spectra (X-band) of correlated radical pairs created in AOT reverse micelles and microemulsions are presented, simulated, and discussed using the microreactor model. The radicals are formed inside the water pool using photooxidation of diglycine by the excited triplet states of two different anthraquinone sulfonate salts. Water pool size and temperature effects on the spectra are reported, and the simulations allow for extraction of the diffusion coefficient in the interior, which monotonically increases with water pool size. The data directly correlate with the diffusional properties of correlated radical pairs in regular aqueous micelle solutions studied previously by similar methods. Competition between H-atom abstraction and electron transfer is observed with anthraquinone sulfonate, but electron transfer is the only reaction pathway observed when anthraquinone disulfonate triplet state is the sensitizing species. PMID- 15779942 TI - Small-angle neutron scattering studies of charged carboxyl-terminated dendrimers in solutions. AB - Small-angle neutron scattering was used to characterize the solution behavior of charged carboxylic acid terminated "cascade" dendrimers (Z-Cascade/methane [4]/3 oxo-6-oxa-2-azaheptylidyne/3-oxo-2-azaheptylidyne/propanoic acids) of third (G3) and fifth (G5) generations as a function of dendrimer concentration, pH, and ionic strength. An increase in dendrimer concentration leads to a single broad peak in the scattering profile arising from interdendrimer interaction. The dissociation of terminal carboxylate groups also gives rise to an interdendrimer interaction peak, which could be suppressed by the addition of excess salt. The results of contrast matching measurements indicate the accumulation of an excess concentration of tetramethylammonium counterions around the surface of these highly charged particles, and the thickness of these counterions lies somewhere between 4 and 6 A. This conclusion is consistent with our previous potentiometric titration (Zhang, H.; et al. J. Phys. Chem. B 1997, 101, 3494) and capillary electrophoresis (Huang, Q. R.; et al. J. Phys. Chem. B 2000, 104, 898) data. PMID- 15779943 TI - Added surfactant can change the phase behavior of aqueous polymer-particle mixtures. AB - The phase behavior of aqueous mixtures of the "clouding" polymer ethyl(hydroxyethyl)cellulose (EHEC) mixed with colloidal particles and surfactants has been studied. These types of mixtures are important in many technical formulations. Two types of particles, polystyrene latex and silica, and two types of EHEC, nonmodified EHEC (N-EHEC) and hydrophobically modified EHEC (HM-EHEC), were studied. The EHECs adsorb to both kinds of particles. Both the amount and the type of added surfactant were seen to dramatically influence the partitioning of the particles between the EHEC-rich and EHEC-poor phases of phase separated mixtures (above the cloud point temperature). Surfactants that are known not to associate with the EHEC backbone, that is, nonionic surfactants and short-chain cationic surfactants, changed the interaction between EHEC and the colloidal particles from attraction to repulsion above a specific surfactant concentration, resulting in a change in the partitioning of the particles from the EHEC-rich to the EHEC-poor phase. No such particle inversion was observed for ionic surfactants that bind to the EHEC backbone. An analysis considering both the binding of surfactant to EHEC and the competitive adsorption of surfactant to the particle surfaces could rationalize all observations, including the large variations observed, among the studied mixtures, in the surfactant concentration required for particle inversion. PMID- 15779944 TI - Distribution of CdSe quantum dots within swollen polystyrene microgel particles using confocal microscopy. AB - CdSe quantum dots (QDs) are semiconducting nanoparticles that fluoresce when stimulated by visible light. This property has been exploited in their use as tracer particles in biomedical applications. In this study, confocal microscopy has been used to determine the distribution of QDs within polystyrene microgel particles, dispersed in an organic solvent. It was found that the extent of microgel swelling affected the penetration of the QDs into the particles. Only when the microgel particles were swollen to their maximum extent were the QDs able to penetrate into the central core region of the particles. PMID- 15779945 TI - Photoinduced gelation by stilbene oxalyl amide compounds. AB - Oxalyl amide derivatives bearing 4-dodecyloxy-stilbene as a cis-trans photoisomerizing unit were synthesized. The trans derivative acted as a versatile gelator of various organic solvents, whereas the corresponding cis derivative showed a poor gelation ability or none at all. In diluted solution (c = 2.0 x10( 5) mol dm(-3), ethanol), the cis isomer was photochemically converted into the trans isomer within 4 min. Depending on the radiation wavelength, the trans isomer was stable or liable to photodecomposition. When exposed to irradiation, a concentrated solution of the cis isomer (c = 2.0 x 10(-2) mol dm(-3), ethanol) turned into a gel. The FT-Raman, FT-IR, and 1H NMR spectra demonstrated that the gelation process occurred because of a rapid cis --> trans photoisomerization followed by a self-assembly of the trans molecules. Apart from the formation of hydrogen bonding between the oxalyl amide parts of the molecules, confirmed by FT IR spectroscopy, it was assumed that the pi-pi stacking between the trans stilbene units of the molecule and a lipophilic interaction between long alkyl chains were the interactions responsible for gelation. PMID- 15779946 TI - Differences in binding of a cationic surfactant to cross-linked sodium poly(acrylate) and sodium poly(styrene sulfonate) studied by Raman spectroscopy. AB - Raman spectroscopy has been used to investigate the structure of gel-surfactant complexes. Cross-linked sodium poly(acrylate) and sodium poly(styrene sulfonate) were immersed in solutions of the cationic surfactant dodecyl trimethylammonium bromide. During the deswelling process, two distinct regions could be observed for both types of gels. Looking at the Raman spectra, however, for the poly(styrene sulfonate), the surfactant could be found throughout the gel particle, whereas for poly(acrylate), essentially all the surfactant was bound in a surface layer. PMID- 15779947 TI - Enhanced adsorption of alkyl glucosides on the silica/water interface by addition of amine oxides. AB - The effect of adding a small amount of dodecyl dimethylamine oxide (DDAO) on adsorption on silica from an aqueous solution of dodecyl maltoside (C12G2) has been investigated. The C12G2 itself does not adsorb significantly on silica at any concentration. DDAO on the other hand readily adsorbs in a bilayer-like structure at concentrations approaching the critical micelle concentration (cmc), but the adsorbed amount at the concentrations it has been applied in these mixtures is small. In contrast, by combination of the two surfactants, significant adsorption is observed at concentrations where the adsorption of the pure DDAO, as well as pure C12G2, is very low. We thus see a strong enhancement of the adsorption from the mixed system. The adsorption is suggested to be a two step process, where individual DDAO molecules first adsorb to the silica surface through electrostatic interactions, and then C12G2 adsorbs at the hydrophobic sites the DDAO tails constitute through hydrophobic bonding. A minimum concentration of DDAO is required to induce adsorption from a solution with constant C12G2 concentration. This concentration is lower for C12G2 solutions below and equal to cmcC12G2 than above cmcC12G2. In addition, the total adsorbed amount shows a maximum around cmcC12G2 of the mixture for a solution with low DDAO contents. Both these effects are explained by incorporation of DDAO in the mixed micelles above cmc(mix), which leads to a desorption of DDAO from the surface. PMID- 15779948 TI - Robust self-assembled octadecylphosphonic acid monolayers on a mica substrate. AB - As determined by scratch tests, self-assembled monolayers (SAMs) of octadecylphosphonic acid (OPA) on a muscovite mica substrate were found to be mechanically robust and to serve as a lubricant to protect the underlying mica substrate. For comparison purposes, three polymer films were subjected to scratch tests under the same conditions. The scratch tests were conducted using a diamond tipped stylus, and the resultant scratches were examined using atomic force microscopy. The excellent mechanical strength of OPA SAMs is supported by analysis with time-of-flight secondary ion mass spectrometry, which suggests that the headgroup of the OPA is strongly bonded to the substrate atoms. The molecular lubrication provided by OPA SAMs suggests that the interaction between the headgroup and the substrate is sufficiently strong to endure significant shear force and that the hydrocarbon chains are able to dissipate shear energy. PMID- 15779949 TI - Surfactant boundary lubricant film modified by an amphiphilic diblock copolymer. AB - The effect of the uptake of a low-molecular-weight amphiphilic diblock copolymer on the morphology of didodecyldimethylammonium bromide (DDAB) adsorbed layers on mica, the interactions between two coated surfaces, and the frictional properties of the boundary film have been studied using an atomic force microscope and a dynamic surface forces apparatus nanotribometer. When DDAB-coated surfaces in aqueous solution were compressed, hemifusion or removal of the adsorbed surfactant bilayers could not be induced, and no frictional force could be measured between the surfaces, which display superior lateral cohesion and lubricant properties. Coadsorbing octadecyl end modified poly(ethylene oxide) chains at low density facilitates hemifusion, generating significant shear stress and leading to stick-slip instabilities. The mixed films regain their lateral cohesion at higher adsorbed copolymer densities, but an extra short-range attraction brings the adsorbed layers into adhesive contact without causing bilayer hemifusion. Here, noticeable frictional forces are also measured. PMID- 15779950 TI - Intriguing self-assembly of large granules of F-actin facilitated by gelsolin and alpha-actinin. AB - We report microscopic observations and a structural determination of actin granules self-assembled in concentrated solutions of actin filaments (F-actin). Optical microscopy shows reproducible formation of numerous and stable granules of densely packed F-actin of variable sizes on the order of 10 microm. These granules coexist with a uniform network of F-actin of a lower concentration. The microscopic segregation of F-actin into two distinct states is assisted by an actin cross-linking protein, alpha-actinin. The rapid on and off rates and temperature sensitivity of the alpha-actinin/F-actin interaction facilitate the formation of multi-micrometer-sized granules of well-defined shapes. Additional physical factors such as the excluded volume effect and the minimization of surface energy act in concert with the specific molecular interactions to define the intriguing granular formation. Both the biochemical specificity of alpha actinin and the thermodynamics of phase transitions are required for understanding such large scale self-assembly. PMID- 15779951 TI - Proton sponge and fatty acid interactions at the air-water interface. Thermodynamic, spectroscopic, and microscopic study. AB - Mixed Langmuir and Langmuir-Blodgett (LB) films of a proton sponge, namely, diphenyl bis(octadecylamino)phosphonium bromide, and a fatty acid, docosanoic acid, with different molar ratios have been fabricated. Surface pressure versus area per molecule isotherms were registered, and the excess areas and excess Gibbs energy of mixing were calculated. Strong interactions between the proton sponge and the fatty acid take place at the air-water interface. The existence of a stoichiometric 1:1 acid-base reaction between the two components forming a complex on the water surface at high surface pressures has been demonstrated. Furthermore, the reaction had an efficiency close to 100% at the air-water interface; meanwhile, it hardly takes place in organic solvents such as chloroform or even a mixture of chloroform and dimethyl sulfoxide. The floating films were transferred to solid supports and characterized by means of several techniques including IR spectroscopy, X-ray diffraction, and atomic force microscopy, revealing the presence of highly ordered alkyl chains and a constant architecture along the different layers as well as the presence of different domains in the LB films, except those having a 1:1 proton sponge-fatty acid ratio that are homogeneous. Such domains have been interpreted as the presence of two different phases, the 1:1 complex plus the excess component in the mixture. PMID- 15779952 TI - Formation of a liquid crystalline phase from phosphatidylcholine at the oil aqueous interface. AB - Adsorption of phospholipid (1,2-dioleoyl-sn-glycero-3-phosphatidylcholine) and formation of a surface phase at the oil-water interface has been followed by using ellipsometry. The properties of the interfacial phase were found to depend strongly on whether phospholipid was added to the oil phase or to the aqueous phase as liposomal structures. In the latter case a monolayer formed, while if the phospholipid was supplied from the oil phase a lamellar phase appeared at the interface. The effect on the stabilizing surface phase of a surface-active protein (beta-lactoglobulin) was also investigated. The observations are important for understanding stabilizing properties of surface-active compounds commonly used to stabilize emulsions. In addition it has been demonstrated that ellipsometry can be used to study the initial process when a two-phase system consisting of a water and an oil phase is transformed into a three phase system or eventually to a one-phase system. PMID- 15779953 TI - Adsorption of organic matter at mineral/water interfaces: 5. Effects of adsorbed natural organic matter analogues on mineral dissolution. AB - The effects of the adsorption of pyromellitate, an analogue for natural organic matter, on the dissolution behavior of corundum (alpha-Al2O3) have been examined over a wide range of pyromellitate concentrations (0-2.5 mM) and pH conditions (2 10). The adsorption modes of pyromellitate on corundum have first been examined using in situ attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy and are shown to be dominated by a fully deprotonated, outer-sphere pyromellitate species ([triple bond]AlOH2+. . .Pyr4-) at pH >/= 5.0. At lower pH conditions, however, an additional protonated outer-sphere species ([triple bond]AlOH2+. . .H2Pyr2-) and an inner-sphere species are also evident. In accordance with the ATR-FTIR findings, modeling of macroscopic pyromellitate adsorption data using an extended constant capacitance treatment was possible using two outer-sphere ([triple bond]AlOH2+. . .Pyr4- and [triple bond]AlOH2+. . .H2Pyr2-) and one inner-sphere ([triple bond]AlPyr3-) adsorbed pyromellitate species. The presence of adsorbed pyromellitate strongly inhibited the dissolution of corundum under acidic (pH < 5) conditions, consistent with a mechanism previously proposed by Johnson et al. whereby outer-spherically adsorbed Pyr4- species sterically protect dissolution-active surface sites from attack by dissolution-promoting species such as protons. A reduction in the protolytic dissolution rate of corundum results. A reference Suwannee River fulvic acid, which also adsorbs to aluminum (oxyhydr)oxide surfaces in a predominantly outer-sphere manner, was similarly shown to strongly inhibit the dissolution of corundum at pH = 3. PMID- 15779954 TI - An experimental and theoretical study of competitive adsorption at the n heptane/water interface. AB - A model to calculate the interfacial concentration of competing surface active species in a two-phase oil/water system was developed. To enable the calculation of the surface excess of 2-hydroxy-5-nonylacetophenone oxime (HNAPO, active ingredient of LIX 84) in the presence of surfactants competing for interfacial area, an interfacial adsorption competition model was derived for noninteracting surface active species in a n-heptane/aqueous system, assuming ideal enthalpy and entropy of mixing. The model was found to be valid for HNAPO in the presence of sodium dodecyl sulfate (SDS) or dodecyldimethyl(3-sulfopropyl)ammonium (DDSA). In the case of dodecyltrimethylammonium chloride (DTAC) or octa(ethylene glycol) mono-n-dodecyl ether (C12E8) as the competing surfactants with HNAPO, the predicted surface excess values from the model fit less favorably. The difference was shown to not be due to nonideal entropy of mixing. PMID- 15779955 TI - Composition of supported model membranes determined by neutron reflection. AB - We have investigated the formation of supported bilayers by coadsorption of dipalmitoyl phosphatidylcholine (DPPC) with the nonionic surfactant beta-D dodecyl maltoside. The adsorption of mixed phospholipid-surfactant micelles on hydrophilic silica surfaces at 25 degrees C was followed as a function of bulk concentration by neutron reflection. Using chain-deuterated d(25)-beta-D-dodecyl maltoside and d(62)-DPPC, we demonstrate that it is possible to determine the composition of the bilayers at each stage of a sequential dilution process, which enriches the adsorbed layer in phospholipid and leads to complete elimination of the surfactant. The final supported bilayers have thicknesses of 51 +/- 3 A and are stable to heating to 37 degrees C once all surfactant has been removed, and the structures agree well with other published data on DPPC supported bilayers. The coadsorption of cholesterol in a DPPC-surfactant mixture was also achieved, and the location and volume fraction of cholesterol in the DPPC bilayer was determined. Cholesterol is located in a 18 +/- 1 A thick layer below the lipid headgroup region and leads to an increased bilayer thickness of 58 +/- 2 A at 26 mol % of cholesterol. PMID- 15779956 TI - Investigation of activated carbon surface heterogeneity by argon and nitrogen low pressure quasi-equilibrium volumetry. AB - Surface heterogeneity can be assessed by adsorption of different gaseous probes on solid materials. In the present study, four types of activated carbons were analyzed by classical N2 Brunauer-Emmett-Teller (BET) measurements and by low pressure quasi-equilibrium volumetry (LPQEV) (Villieras, F.; Michot, L. J.; Bardot, F.; Cases, J. M.; Francois, M.; Rudzinski, W. Langmuir 1997, 13, 1104). Three methods of data evaluation were applied: (a) the Frenkel-Halsey-Hill method for estimation of fractal dimensions from BET data, (b) the Horwath-Kawazoe method to calculate the pore size distribution from LPQEV Ar and N2 adsorption isotherms, and (c) the derivative isotherm summation (DIS) method to describe the solid's surface heterogeneity by a concept of local derivative isotherms. Similar Ar and N2 adsorption energy distributions were obtained on all carbons, which indicates the presence of mainly nonpolar surfaces. When adsorption was described by the van der Waals equation, the ratio between the interaction energy of different energetic sites with argon and nitrogen was 0.88. This value corresponded very well with a slope obtained when Ar and N2 positions of local isotherms by the DIS method were compared. This relationship has an important impact because it enables one to constrain the modeling of local isotherms. This study, besides the surface information, showed large possibilities of the DIS method for the surface analysis not only in terms of solid heterogeneity characterization but also in terms of polarity assessment. PMID- 15779957 TI - Analysis of desiccation and vitrification characteristics of carbohydrate films by shear-wave resonators. AB - Desiccated state preservation of mammalian cells and tissues in the presence of carbohydrates has started to show promise in the last two decades. Certain carbohydrates play a major role in preservation by reducing molecular mobility in the desiccated state. In this communication, the feasibility of utilizing shear wave resonators to collect real-time molecular mobility information during desiccation and vitrification of carbohydrate based thin films was demonstrated. Simultaneous quartz crystal microbalance experimentation and optical imaging were utilized to determine the conditions for thin film formation and the vitrification kinetics of certain carbohydrate solutions of biological importance. Using the technique presented here, it was possible to gain insight into the vitrification characteristics of carbohydrate solutions establishing the basics for future research with quantitative analysis of film properties and experimentation with live mammalian cells. PMID- 15779958 TI - Adsorption and aggregation of cationic amphiphilic polyelectrolytes on silica. AB - The adsorption of two cationic amphiphilic polyelectrolytes, which are copolymers of two charged monomers, triethyl(vinylbenzyl)ammonium chloride and dimethyldodecyl(vinylbenzyl)ammonium chloride (which is the amphiphilic one) with different contents of amphiphilic groups (40% (40DT) and 80% (80DT)), onto the hydrophilic silica-aqueous solution interface has been studied by in situ null ellipsometry and tapping mode atomic force microscopy (AFM). Adsorption isotherms for both polyelectrolytes were obtained at 25 degrees C and at different ionic strengths, and the adsorption kinetics was also investigated. At low ionic strength, thin adsorbed layers were observed for both polyelectrolytes. The adsorption increases with polymer concentration and reaches, in most cases, a plateau at a concentration below 50 ppm. For the 80DT polymer, at higher ionic strength, an association into aggregates occurs at concentrations at and above 50 ppm. The aggregates were observed directly by AFM at the surface, and by dynamic light scattering in the solution. The adsorption data for this case demonstrated multilayer formation, which correlates well with the increase in viscosity with the ionic strength observed for 80DT. PMID- 15779959 TI - Neutron reflectivity study of lipid membranes assembled on ordered nanocomposite and nanoporous silica thin films. AB - Single bilayer membranes of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were formed on ordered nanocomposite and nanoporous silica thin films by fusion of small unilamellar vesicles. The structure of these membranes was investigated using neutron reflectivity. The underlying thin films were formed by evaporation induced self-assembly to obtain periodic arrangements of silica and surfactant molecules in the nanocomposite thin films, followed by photocalcination to oxidatively remove the organics and render the films nanoporous. We show that this platform affords homogeneous and continuous bilayer membranes that have promising applications as model membranes and sensors. PMID- 15779960 TI - Adsorption of nitrogen on granular activated carbon: experiment and modeling. AB - A carbon adsorbent was produced and used to volumetrically measure nitrogen adsorption isotherms from 93 to 298 K and up to 7 MPa. The isosteric heat of adsorption was determined to range between -9.5 and -16 kJ/mol. The excess adsorption isotherms were modeled using an approach based on a modified Dubinin Astakhov adsorption model, adapted for excess adsorption, which provided an accurate fit for all supercritical isotherms. An expression for the differential energy of adsorption as a function of pressure was developed using the Dubinin Astakhov isotherm. The energy of adsorption for the isotherms measured was found to range from -8 to -15 kJ/mol as a function of pressure. PMID- 15779961 TI - Templated surfactant readsorption on polyelectrolyte-induced depleted surfactant surfaces. AB - Changes in the structure of a surfactant adsorbed on oxidized silicon arising from interaction with a polyelectrolyte have been studied using polarized infrared attenuated total reflection spectroscopy. Specifically, the cationic surfactant cetyltrimethylammonium bromide (CTAB) was found to form a highly ordered layer on oxidized silicon at a concentration of 5.5 x 10(-5) M and a pH of 9.6. Addition of a solution of the anionic polyelectrolyte poly(styrenesulfonate) to the ordered CTAB layer resulted in a rapid and dramatic decrease in the surface excess of CTAB. Interestingly however, the interfacial order of the residual surfactant was retained for a time period greater than 1 h, before decreasing. Reintroduction of a surfactant solution prior to destabilization of the residual interfacial CTAB resulted in the readsorption of the surfactant, the recovery of the initial equilibrium coverage, and the maintenance of an ordered CTAB conformation. This desorption/readsorption process may be subsequently repeated without destroying the order of the CTAB on the surface. If however sufficient time is allowed for the residual interfacial surfactant to destabilize prior to readdition of CTAB, the degree of surfactant order remains low, despite the rapid reobtainment of a surface excess equal to or greater than that initially measured. These results are interpreted in terms of polymer/surfactant interfacial complexation and the removal of adsorbed surfactant into solution. The ordering behavior of the residual surfactant suggests that CTAB is left on the surface in isolated patches of highly ordered species that maintain their order until two-dimensional diffusion leads to a more homogeneous surfactant surface distribution and hence the loss of conformational order. The degree of orientation order assumed by surfactant readsorbing on a depleted surface appears to be templated by the order of the residual interfacial surfactant, suggestive of a two-dimensional epitaxial growth mechanism for CTAB readsorption. PMID- 15779962 TI - Instabilities during the formation of electroactive polymer thin films. AB - The solvent-induced film structure of poly(n-vinyl carbazole) (PVK) thin films on indium tin oxide (ITO)-coated glass was examined. PVK thin films were prepared via spin-coating using five different solvents. We investigated the relationship between the solvent characteristics and film properties, including surface roughness and structure, film thickness, and density. The spin-coated polymer thin films are not in thermodynamic equilibrium; rather, the film properties are affected by the dynamics of the spin-coating process. We found that water present in tetrahydrofuran (THF) induces dewetting of PVK films during the spin-coating process. Solvents with a high evaporation rate lead to high surface roughness due to Marangoni convection. The results show that the surface roughness and structure of the films are dominated by the dynamics of the film formation process, rather than thermodynamic interactions between the polymer and solvents. PMID- 15779963 TI - Effect of surface area on equilibrium pressure. Adsorption approach. AB - Changing droplet radii in a liquid-vapor system is due to the phase transition on the droplet surface. As a variation of the internal energy does not depend on the way the change occurs, we can imagine that a gas condenses on a droplet surface in two stages: in the first stage, autoadsorption occurs on the liquid surface, and in the second stage, adsorbed molecules transfer into the volume by diffusion. Assuming that the energetic effects of the diffusion are independent of the surface curvature, one may conclude that if two liquid bodies differ only with respect to their geometry, the difference of enthalpies of condensation on their surfaces, DeltaH(bd), is equal to the variation of energies of autoadsorption. An estimation of DeltaH(bd) for the simple bodies is presented, and the relationship between the saturation pressure and droplet radii is derived. In the range of micrometer dimensions, the new equation and the Kelvin model lead to close results; for nanocapillaries, the Kelvin equation predicts a divergence of hysteresis loops, whereas the new equation adequately describes the observations. The classical model presumes that a surface area, A, affects the free energy, while the new approach is based on the assumption that A is the repository for the internal energy. PMID- 15779964 TI - Slow adsorption reaction between arsenic species and goethite (alpha-FeOOH): diffusion or heterogeneous surface reaction control. AB - The slow stage of phosphate or arsenate adsorption on hydrous metal oxides frequently follows an Elovich equation. The equation can be derived by assuming kinetic control by either a diffusion process (either interparticle or intraparticle) or a heterogeneous surface reaction. The aim of this study is to determine whether the slow stage of arsenic adsorption on goethite is more consistent with diffusion or heterogeneous surface reaction control. Adsorption kinetics of arsenate and dimethylarsinate (DMA) on goethite (alpha-FeOOH) were investigated at different pH values and inert electrolyte concentrations. Their adsorption kinetics was described and compared using Elovich (Gamma vs ln time) plots. Desorption of arsenate and DMA was studied by increasing the pH of the suspension from pH 4.0 to pH 10.0 or 12.0. The effective particle sizes and zeta potential of goethite were also determined. Effective particle size increased rapidly as the pH approached pH(IEP), both in the absence and presence of arsenic. Inert electrolyte concentrations and pH had no effect on the slow stage of arsenate adsorption on goethite, while the kinetics of DMA adsorption on goethite was influenced by both parameters. The slow stage of arsenate adsorption on goethite follows an Elovich equation. Since effective particle size changes with both pH and inert electrolyte concentrations, and effective particle size influences interparticle diffusion, the arsenate adsorption kinetics indicate that the slow adsorption step is not due to interparticle diffusion. DMA also has complex adsorption kinetics with a slow adsorption stage. DMA desorbed completely and rapidly when the pH was raised, in contrast to the slow adsorption kinetics, indicating that the slow adsorption step is not due to intraparticle diffusion. The slow adsorption is not the result of diffusion, but rather is due either to the heterogeneity of the surface site bonding energy or to other reactions controlling arsenic removal from solution. PMID- 15779965 TI - Systematic control of the packing density of self-assembled monolayers using bidentate and tridentate chelating alkanethiols. AB - The structural and interfacial properties of self-assembled monolayers (SAMs) on gold derived from the adsorption of a series of 1,1,1-tris(mercaptomethyl)alkanes (i.e., CH3(CH2)mC[CH2SH]3, where m = 9, 11, 13, 15) were investigated. The new SAMs, which possess uniformly low densities of alkyl chains, were characterized by ellipsometry, contact angle goniometry, and polarization modulation infrared reflection absorption spectroscopy. Additional analysis of the SAMs by X-ray photoelectron spectroscopy permitted a direct calculation of the packing densities of the SAMs on gold. The results as a whole, when compared to those obtained on SAMs generated from normal alkanethiols (CH3(CH2)m+2SH), 2 alkylpropane-1,3-dithiols (CH3(CH2)mCH[CH2SH]2), and 2-alkyl-2-methylpropane-1,3 dithiols (CH3(CH2)mC(CH3)[CH2SH]2) having analogous chain lengths, demonstrate that the 1,1,1-tris(mercaptomethyl)alkanes afford SAMs with alkyl chains having the lowest packing density and least conformational order. PMID- 15779966 TI - Measurement of dynamical forces between deformable drops using the atomic force microscope. I. Theory. AB - Recent experimental developments have enabled the measurement of dynamical forces between two moving liquid drops in solution using an atomic force microscope (AFM). The drop sizes, interfacial tension, and approach velocities used in the experiments are in a regime where surface forces, hydrodynamics, and drop deformation are all significant. A detailed theoretical model of the experimental setup which accounts for surface forces, hydrodynamic interactions, droplet deformation, and AFM cantilever deflection has been developed. In agreement with experimental observations, the calculated force curves show pseudo-constant compliance regions due to drop flattening, as well as attractive pull-off forces due mainly to hydrodynamic lubrication forces. PMID- 15779967 TI - Self-assembly patterns formed upon solvent evaporation of aqueous cetyltrimethylammonium bromide-coated gold nanoparticles of various shapes. AB - Gold nanocrystals of various shapes, which were produced in high yield in the presence of cetyltrimethylammonium bromide (CTAB), showed a range of two dimensional self-assembly patterns upon drying from aqueous solution. The interparticle spacings were independent of the size and shape of the gold nanocrystals. Energy-dispersive X-ray analysis (EDAX) and Fourier-transform infrared (FTIR) spectroscopic studies revealed that the CTAB molecules adsorb onto surfaces of the gold nanocrystals in a bilayer or multilayer fashion, consistent with other groups' results. Zeta potential measurements showed that CTAB-coated nanocrystals were positively charged and the zeta potential remained almost the same upon two centrifugations and redispersion of the nanocrystals in deionized water, confirming the high stability of the surfactant-nanoparticle interaction. The nanocrystal shape strongly influenced the nature of the self assembly patterns, in some cases in accord with theoretical predictions. CTAB is proposed as the medium for self-assembly, via interdigitation of its hydrophobic chains from adjacent nanocrystals for close contact, or via sharing a layer of counterions for larger inter-nanocrystal spacings. PMID- 15779968 TI - Adsorption of poly(ethylene oxide)-b-poly(epsilon-caprolactone) copolymers at the silica-water interface. AB - The adsorption of amphiphilic poly(ethylene oxide)-b-poly(epsilon-caprolactone) and poly(ethylene oxide)-b-poly(gamma-methyl-epsilon-caprolactone) copolymers in aqueous solution on silica and glass surfaces has been investigated by flow microcalorimetry, small-angle neutron scattering (SANS), surface forces, and complementary techniques. The studied copolymers consist of a poly(ethylene oxide) (PEO) block of M(n) = 5000 and a hydrophobic polyester block of poly(epsilon-caprolactone) (PCL) or poly(gamma-methyl-epsilon-caprolactone) (PMCL) of M(n) in the 950-2200 range. Compared to homoPEO, the adsorption of the copolymers is significantly increased by the connection of PEO to an aliphatic polyester block. According to calorimetric experiments, the copolymers interact with the surface mainly through the hydrophilic block. At low surface coverage, the PEO block interacts with the surface such that both PEO and PCL chains are exposed to the aqueous solution. At high surface coverage, a dense copolymer layer is observed with the PEO blocks oriented toward the solution. The structure of the copolymer layer has been analyzed by neutron scattering using the contrast matching technique and by tapping mode atomic force microscopy. The experimental observations agree with the coadsorption of micelles and free copolymer chains at the interface. PMID- 15779969 TI - Surface modification of polyacrylonitrile-based membranes by chemical reactions to generate phospholipid moieties. AB - A novel approach for the surface modification of poly(acrylonitrile-co-2 hydroxyethyl methacrylate) (PANCHEMA) membranes by introducing phospholipid moieties is presented, which involved the reaction of the hydroxyl groups on the membrane surface with 2-chloro-2-oxo-1,3,2-dioxaphospholane (COP) followed by the ring-opening reaction of COP with trimethylamine. The chemical changes of phospholipid-modified acrylonitrile-based copolymers (PMANCP) membranes were characterized by Fourier transfer infrared spectroscopy and X-ray photoelectron spectroscopy. The surface properties of PMANCP membranes were evaluated by pure water contact angle, protein adsorption, and platelet adhesion measurements. Pure water contact angles measured by the sessile drop method on PMANCP membranes were obviously lower than those measured on the PANCHEMA membranes and decreased with the increase of the content of phospholipid moieties on the membrane surface. It was found that the bovine serum albumin adsorption and platelet adhesion were suppressed significantly with the introduction of phospholipid moieties on the membranes surface. These results demonstrated that the described process was an efficient way to improve the surface biocompatibility for the acrylonitrile-based copolymer membrane. PMID- 15779970 TI - Texture changes inside smectic-C droplets in azobenzene Langmuir monolayers. AB - Preparation of Langmuir monolayers of a mixture of trans- and cis-isomers of an azobenzene derivative, 4-[4-[(4-octylphenyl)azo]phenoxy]butanoic acid, results in the segregation of birefringent trans-isomer domains embedded in an isotropic medium of cis-isomers. Brewster angle microscopy observations allow us to identify different textures inside the domains depending on surface pressure, temperature, and domain size. The evolution of the monolayer in the dark, from initial droplets formed after spreading to a stable stripe texture, is described. The dynamics of domain coalescence and some morphological transitions induced by temperature and surface pressure changes are also discussed. A simple theoretical model is included to supplement some of these experimental observations. PMID- 15779971 TI - Probing of porphyrin surface chemistry in systems with laser-ablated Ag nanoparticle hydrosol: role of thiosulfate anions. AB - The influence of sodium thiosulfate (THS) concentration in Ag colloid/THS/H(2)TMPyP and Ag colloid/H2TMPyP/THS systems (H2TMPyP = 5,10,15,20 tetrakis(1-methyl-4-pyridyl)porphyrin) was investigated by a combination of surface-enhanced resonance Raman scattering (SERRS) spectroscopy, surface plasmon extinction (SPE) measurements, and transmission electron microscopy (TEM). THS was found to have a strong impact on Ag nanoparticle surface structure and aggregation state and on interaction with H2TMPyP probe molecules, as evidenced by variations of the SERRS spectrum. In the Ag colloid/THS/H2TMPyP system, when laser-ablated Ag colloid was THS pretreated prior to the porphyrin addition, a critical threshold THS concentration (4 x 10(-5) M) was discovered. At concentrations below the threshold, THS mainly reduces the number of Ag+ adsorption sites. This leads to increased Ag nanoparticle aggregation prior to the porphyrin addition and significant weakening of the overall SERRS signal. Dominant contributions in the SERRS spectrum correspond to free base H2TMPyP and Ag+ containing the AgTMPyP form. At concentrations above the threshold, THS mediates also the formation and stabilization of new adsorption sites, probably Ag(0) sites. This induces a turn in the aggregation state of the pretreated Ag c/THS system, an increase of the overall SERRS signal, and the appearance of a new spectral form of Ag metalated porphyrin. PMID- 15779972 TI - Self-assembled shells composed of colloidal particles: fabrication and characterization. AB - We construct shells with tunable morphology and mechanical response with colloidal particles that self-assemble at the interface of emulsion droplets. Particles self-assemble to minimize the total interfacial energy, spontaneously forming a particle layer that encapsulates the droplets. We stabilize these layers to form solid shells at the droplet interface by aggregating the particles, connecting the particles with adsorbed polymer, or fusing the particles. These techniques reproducibly yield shells with controllable properties such as elastic moduli and breaking forces. To enable diffusive exchange through the particle shells, we transfer them into solvents that are miscible with the encapsulant. We characterize the mechanical properties of the shells by measuring the response to deformation by calibrated microcantilevers. PMID- 15779973 TI - Synthesis and self-assembly of galactose-terminated alkanethiols and their ability to resist proteins. AB - The synthesis of two galactose-terminated alkanethiols with the structural formula X-OC2H5NHCO(CH2)15SH (X = 2,3,4,6-tetra-O-methyl-beta-D-Gal or beta-D Gal) is described. Single-component and mixed self-assembled monolayers (SAMs) of the methylated and nonmethylated compounds were prepared on gold and subsequently characterized with ellipsometry, contact angle goniometry, and infrared reflection-absorption spectroscopy. Studies of the irreversible protein adsorption onto the SAMs using ex-situ ellipsometry revealed very low levels of fibrinogen and lysozyme adsorption onto mixed SAMs displaying advancing water contact angles between 24 degrees and 45 degrees and below 45 degrees , respectively. A monomethylated compound (X = 6-O-methyl-beta-D-Gal) was also synthesized and assembled on gold. This particular compound was found to possess wettability properties corresponding to the low adsorption regime of the mixed SAMs, and the results from the same set of fibrinogen and lysozyme adsorption experiments showed very low levels of protein adsorption. Our findings suggest that the protein rejecting properties rely on a fine balance between the surface energy and/or hydrogen bond donating/accepting properties of the SAM surface. PMID- 15779974 TI - Poly(ethylene glycol) self-assembled monolayer island growth. AB - Here, we report a study of the morphology and growth dynamics of a self-assembled monolayer (SAM) of the amide containing poly(ethylene glycol) (PEG) thiol (CH3O(CH2CH2O)17NHCO(CH2)2SH) on atomically flat Au(111) surfaces. SAM growth from a 20 muM ethanolic solution reveals island growth through three distinct steps: island nucleation, island growth, and coalescence. The coalescence-step, filling voids in the SAM, is by far slowest. The fine structure study reveals dendritic island formation, an observation which can be explained by attractive intermolecular interactions and surface diffusion-limited aggregation. We have also observed a change in the island height, which peaks during the island growth phase. This height change can be associated with a molecular conformational transition. PMID- 15779975 TI - Kinetics of salicylic acid adsorption on activated carbon. AB - The adsorption and desorption of salicylic acid from water solutions was investigated in HPLC microcolumns packed with activated carbon. The adsorption isotherm was obtained by the step-up frontal analysis method in a concentration range of 0-400 mg/L and was well fitted with the Langmuir equation. The investigation of rate aspects of salicylic acid adsorption was based on adsorption/desorption column experiments where different inlet concentrations of salicylic acid were applied in the adsorption phase and desorption was conducted with pure water. The concentration profiles of individual adsorption/desorption cycles data were fitted using several single-parameter models of the fixed-bed adsorption to assess the influence of different phenomena on the column behavior. It was found that the effects of axial dispersion and extraparticle mass transfer were negligible. A rate-determining factor of fixed-bed column dynamics was the kinetics of pore surface adsorption. A bimodal kinetic model reflecting the heterogeneous character of adsorbent pores was verified by a simultaneous fit of the column outlet concentration in four adsorption/desorption cycles. The fitted parameters were the fraction of mesopores and the adsorption rate constants in micropores and mesopores, respectively. It was shown that the former rate constant was an intrinsic one whereas the latter one was an apparent value due to the effects of pore blocking and diffusional hindrances in the micropores. PMID- 15779976 TI - Molecular rectification: application in dye-sensitized solar cells. AB - A dye-sensitized heterojunction of configuration n-TiO2/PD-CuPC-MV/p-CuSCN (where PD = 3,4-pyridinedicarboxylic acid anchored to TiO2, CuPC = copper(II) phthallocyanine tetrasulfonic acid ionically linked to PD, and MV = Methyl Violet complexed to CuPC) is developed to demonstrate the applicability of molecular rectification to dye-sensitized solar cells as a strategy of suppressing recombination. Short-circuit photocurrent, open-circuit voltage, energy conversion efficiency, and incident photon to photocurrent conversion of this system are higher than that of the heterojunctions of configurations n-TiO2/PD MV/p-CuSCN, n-TiO/CuPC-MV/p-CuSCN, and n-TiO2/MV/p-CuSCN. The impressively high rectification ratio and the mode of anchorage of CuPC toTiO2 are suggested as the cause of superior photovoltaic action of the cell TiO2/PD-CuPC-MV/p-CuSCN. PMID- 15779977 TI - Solid-state NMR structural studies of peptides immobilized on gold nanoparticles. AB - In this paper we describe solid-state NMR experiments that provide information on the structures of surface-immobilized peptides. The peptides are covalently bound to alkanethiolates that are self-assembled as monolayers on colloidal gold nanoparticles. The secondary structure of the immobilized peptides was characterized by quantifying the Ramachandran angles phi and psi. These angles were determined in turn from distances between backbone carbonyl 13C spins, measured with the double-quantum filtered dipolar recoupling with a windowless sequence experiment, and by determination of the mutual orientation of chemical shift anisotropy tensors of 13C carbonyl spins on adjacent peptide planes, obtained from the double-quantum cross-polarization magic-angle spinning spectrum. It was found that peptides composed of periodic sequences of leucines and lysines were bound along the length of the peptide sequence and displayed a tight alpha-helical secondary structure on the gold nanoparticles. These results are compared to similar studies of peptides immobilized on hydrophobic surfaces. PMID- 15779978 TI - Near-IR light-sensitized voltaic conversion system using nanocrystalline TiO2 film by Zn chlorophyll derivative aggregate. AB - A Zn chlorophyll-a derivative, Zn chlorin-e6 (ZnChl-e6), adsorbed onto a nanocrystalline TiO2 film (ZnChl-e6/TiO2) electrode was prepared, and the photovoltaic properties of the ZnChl-e6/TiO2 electrode were studied. The absorption peaks of ZnChl-e6/TiO2 observed at 420, 654, and 795 nm were attributed to the ZnChl-e6 molecules aggregating onto TiO2 film. The fluorescence attributed to the ZnChl-e6 monomer and aggregate was observed at 710 and 820 nm, respectively, and the fluorescence in both cases was quenched by TiO2 particles. The maximum of the incident photon-to-current conversion efficiency (IPCE) value in the photocurrent action spectrum was 800 nm, and the IPCE value was 7.0%. ZnChl-e6 molecules formed aggregates on a nanocrystalline TiO2 film electrode. From the photocurrent-photovoltage characteristics of the ZnChl-e6/TiO2 electrode irradiated with 100 mW cm(-2), the short-circuit photocurrent (I(SC)) was found to be 0.19 mA cm(-2) and the open-circuit photovoltage (V(OC)) was found to be 375 mV. The maximum power was estimated to be 28.7 microW cm(-2), and the fill factor (FF) was estimated to be 40.1%. A near-IR light induced photovoltaic conversion system using a ZnChl-e6 aggregate formed onto a nanocrystalline TiO2 film electrode was achieved. PMID- 15779979 TI - Preparation, characterization, and electrocatalytic activity of surface anchored, Prussian Blue containing starburst PAMAM dendrimers on gold electrodes. AB - Gold bead electrodes were modified with submonolayers of 3-mercaptopropionic acid or 2-aminoethanethiol and further reacted with poly(amidoamine) (PAMAM) dendrimers (generation 4.0 and 3.5, respectively) to obtain films on which Prussian Blue (PB) was later absorbed to afford mixed and stable electrocatalytic layers. Experiments carried out with these novel materials not only showed an improved surface coverage of PB on the dendrimer modified electrodes as compared to PB modified gold electrodes prepared under acidic conditions, but also showed an increased stability at neutral pH values for one of the dendrimer containing substrates where the PB film on a bare gold electrode is simply not formed. The dendrimer modified electrodes were also tested as electrocatalytic substrates for the electroxidation of L(+)-ascorbic acid (AA), and it was found that their sensitivity as well as the corresponding detection limits were improved as compared to the voltammetric response of a Au-PB modified electrode. On the basis of UV-visible (UV-vis) spectroscopy and electrochemical experiments, it is suggested that the PB molecules are located within the dendritic structure of the surface attached PAMAM dendrimers. PMID- 15779980 TI - Indium tin oxide electrodes modified with tris(2,2'-bipyridine-4,4'-dicarboxylic acid) iron(II) and the catalytic oxidation of tris(4,4'-di-tert-butyl-2,2' bipyridine) cobalt(II). AB - Indium tin oxide (ITO) electrodes modified by attachment of tris(2,2'-bipyridine 4,4'-dicarboxylic acid) iron(II) are examined. The mode of attachment is believed to be via the COOH functions in a manner similar to attachment of similar carboxylate-containing compounds to TiO2 surfaces. On the surface the complex resides as a stable electrochemically active monolayer. These modified electrodes can efficiently catalyze the oxidation of certain cobalt complexes, specifically, tris(4,4'-di-tert-butyl-2,2'-bipyridine) cobalt(II). On the unmodified ITO surfaces this cobalt complex is essentially electrochemically inert. The catalytic process approaches diffusional control at very slow scan speeds. Also, the electro-catalysis is sufficiently efficient that the peak oxidation current for Co2+, under certain conditions, exceeds the i(p) for the surface oxidation of the adsorbed Fe2+ by >x100 and the current for the uncatalyzed oxidation of Co2+ by considerably more than that. PMID- 15779981 TI - 29Si NMR study of structural ordering in aluminosilicate geopolymer gels. AB - A systematic series of aluminosilicate geopolymer gels was synthesized and then analyzed using 29Si magic-angle spinning nuclear magnetic resonance (MAS NMR) in combination with Gaussian peak deconvolution to characterize the short-range ordering in terms of T-O-T bonds (where T is Al or Si). The effect of nominal Na2O/(Na2O + K2O) and Si/Al ratios on short-range network ordering was quantified by deconvolution of the 29Si MAS NMR spectra into individual Gaussian peaks representing different Q4(mAl) silicon centers. The deconvolution procedure developed in this work is applicable to other aluminosilicate gel systems. The short-range ordering observed here indicates that Loewenstein's Rule of perfect aluminum avoidance may not apply strictly to geopolymeric gels, although further analyses are required to quantify the degree of aluminum avoidance. Potassium geopolymers appeared to exhibit a more random Si/Al distribution compared to that of mixed-alkali and sodium systems. This work provides a quantitative account of the silicon and aluminum ordering in geopolymers, which is essential for extending our understanding of the mechanical strength, chemical and thermal stability, and fundamental structure of these systems. PMID- 15779982 TI - Mechanism of selective oxygen reduction on platinum by 2,2'-bipyridine in the presence of methanol. AB - Mechanism of selective oxygen reduction on platinum by 2,2'-bipyridine in the presence of methanol has been investigated by in situ surface-enhanced infrared absorption spectroscopy. The addition of 2,2'-bipyridine caused the decrease of adsorbed water molecules and those existing near the surface of platinum. The formation of both CO and formate, the latter being the intermediate in the non-CO path for methanol oxidation, depressed in the presence of 2,2'-bipyridine, suggests that 2,2'-bipyridine hinders methanol oxidation via both non-CO and CO paths on platinum. The geometrical effect of 2,2'-bipyridine adsorbed onto platinum was also investigated by multisite Monte Carlo simulation. It is indicated that selective oxygen reduction is caused by the difference in the number of required adsorption sites between methanol and dioxygen molecules. The suppression of Pt oxide species by 2,2'-bipyridine is found to be another factor that enhances the oxygen reduction. PMID- 15779983 TI - The antifouling and fouling-release performance of hyperbranched fluoropolymer (HBFP)-poly(ethylene glycol) (PEG) composite coatings evaluated by adsorption of biomacromolecules and the green fouling alga Ulva. AB - Cross-linked hyperbranched fluoropolymer (HBFP) and poly(ethylene glycol) (PEG) amphiphilic networks with PEG weight percentages of 14% (HBFP-PEG14), 29% (HBFP PEG29), 45% (HBFP-PEG45), and 55% (HBFP-PEG55) were prepared on 3 aminopropyl)triethoxysilane (3-APS) functionalized microscope glass slides for marine antifouling and fouling-release applications. The surface-free energies (gamma(s)), polar (gamma(s)(p) and gamma(s)(AB)), and dispersion (gamma(s)(d) and gamma(s)(LW)) components were evaluated using advancing contact angles by two liquid geometric-mean and three-liquid Lifshitz-van der Waals acid-base approaches. The HBFP coating exhibited a low surface energy of 22 mJ/m(2), while the gamma(s) and gamma(s)(p) of the cross-linked HBFP-PEG coatings increased proportionally with the PEG weight percentages in the networks. The adsorption of bovine serum albumin (BSA), lectin from Codium fragile (CFL), lipopolysaccharides from Escherichia coli (LPSE) and Salmonella minnesota (LPSS) upon glass, APS glass, HBFP, PEG, and the cross-linked HBFP-PEG network coatings were investigated by fluorescence microscopy. The marine antifouling and fouling release properties of the cross-linked HBFP-PEG coatings were evaluated by settlement and release assays involving zoospores of green fouling alga Ulva (syn. Enteromorpha; Hayden, H. S.; Blomster, J.; Maggs, C. A.; Silva, P. C.; Stanhope, M. J.; Waaland, J. R. Eur. J. Phycol. 2003, 38, 277). The growth and release of Ulva sporelings were also investigated upon the HBFP-PEG45 coating in comparison to a poly(dimethylsiloxane) elastomer (PDMSE) standard material. Of the heterogeneous cross-linked network coatings, the maximum resistances to protein, lipopolysaccharide, and Ulva zoospore adhesion, as well as the best zoospore and sporeling release properties, were recorded for the HBFP-PEG45 coating. This material also exhibited better performance than did a standard PDMSE coating, suggesting its unique applicability in fouling-resistance applications. PMID- 15779984 TI - Influence of antibody immobilization strategy on molecular recognition force microscopy measurements. AB - A systematic evaluation of the effects of antibody immobilization strategy on the binding efficiency and selectivity (e.g., ability to distinguish between specific and nonspecific interactions) of immunosurfaces prepared with F(ab') antibody fragments of rabbit Immunoglobulin G (IgG) is described. F(ab') was attached to gold surfaces either (1) directly via the formation of a gold-thiolate bond or (2) indirectly through a series of a bifunctional linkers containing an alkane chain or ethylene glycol spacer. Immobilization of F(ab') via the sulfhydryl reactive group located opposite the antigen binding site ensured optimum orientation of the antigen binding site. X-ray photoelectron spectroscopy (XPS) and surface plasmon resonance (SPR) were used to confirm surface modification with the bifunctional linkers and antibody immobilization, respectively. Binding efficiency assays performed with SPR indicated that increasing the length of the linker increased the antigen binding efficiency. Atomic force microscopy (AFM) adhesion force measurements indicated that AFM probes functionalized with directly immobilized F(ab') more effectively discriminated between specific and nonspecific surface-bound proteins than probes modified indirectly via linker immobilized F(ab'). In addition, a greater number of antibody-antigen binding events were observed with directly immobilized F(ab')-functionalized probes. PMID- 15779985 TI - Regiospecific control of protein expression in cells cultured on two-component counter gradients of extracellular matrix proteins. AB - This work describes the use of microfluidic tools to generate covalently immobilized counter gradients of extracellular matrix (ECM) proteins laminin and collagen I. Using these platforms, we demonstrate control of the expression levels of two proteins linked to cell cycle progression by virtue of the spatial location of cells on the gradients, and hence by the local ECM environments in these devices. In contrast to physisorbed gradients, covalently immobilized protein patterns preserved the gradient fidelity, making long term cell studies feasible. This method of precisely controlling local cell environments is simple and broadly portable to other cell types and to other ECM proteins or soluble factors. Our approach promises to enable new investigations in cell biology that will contribute to the establishment of biological design rules for controlling cell growth, differentiation, and function. PMID- 15779986 TI - Near-IR absorption and fluorescence spectra and AFM observation of the light harvesting 1 complex on a mica substrate refolded from the subunit light harvesting 1 complexes of photosynthetic bacteria Rhodospirillum rubrum. AB - The subunit light-harvesting 1 (LH 1) complexes isolated from photosynthetic bacteria Rhodospirillum rubrum using n-octyl-beta-glucoside were reassociated and adsorbed on a mica substrate using spin-coat methods with the aim of using this LH complex in a nanodevice. The near-IR absorption and fluorescence spectra of the LH 1 complexes indicated that the LH 1 complex on the mica was stable, and efficient energy transfer from a carotenoid to a bacteriochlorophyll a was observed. Atomic force microscopy of the reassociated LH 1 complexes, under air, showed the expected ringlike structure. The outer and inner diameters of the ringlike structure of the LH 1 complex were approximately 30 and 8 nm, respectively, and the ringlike structure protruded by 0.2-0.6 nm. PMID- 15779987 TI - Ultrathin coatings from isocyanate terminated star PEG prepolymers: patterning of proteins on the layers. AB - This study presents the easy and fast patterning of low molecular weight molecules that act as binding partners for proteins on Star PEG coatings. These coatings are prepared from isocyanate terminated star shaped prepolymers and form a highly cross-linked network on the substrate in which the stars are connected via urea groups and free amino groups are present. Streptavidin has been patterned on these layers by microcontact printing (muCP) of an amino reactive biotin derivative and consecutive binding of streptavidin to the biotin. Patterns of Ni(2+)-nitriltriacetic acid (NTA) receptors have been prepared by printing amino functional NTA molecules in freshly prepared Star PEG layers that still contain amino reactive isocyanate groups. Complexation of the NTA groups with Ni(II) ions enabled the binding of His-tag enhanced green fluorescent protein (EGFP) in the desired pattern on the substrates. Since the unmodified Star PEG layers prevent unspecific protein adsorption, His-EGFP could selectively be bound to the sample by immersion into crude, nonpurified His-tag EGFP containing cell lysate. PMID- 15779988 TI - Probe surface chemistry dependence and local polymer network structure in F-actin microrheology. AB - We investigate the dependence of F-actin microrheology on probe surface chemistry using diffusing wave spectroscopy. Polystyrene probe particles exhibit subdiffusive mean-squared displacements, where Deltar(2)(t) approximately t(0.77)(+/-)(0.03) consistent with previous experiments and theory. However, polystyrene probes preadsorbed with bovine serum albumin (BSA) interact weakly with the surrounding polymer network and exhibit a scaling exponent similar to pure diffusion Deltar(2)(t) approximately t, which decreases as particle size and actin concentration increases. Using models of particle diffusion in locally heterogeneous viscoelastic microenvironments, we find that the microrheological response of BSA-treated particles is consistent with the formation of a polymer depleted shell surrounding the probes. The shell thickness scales with particle size but not polymer concentration. These results suggest that the depletion is caused by exclusion or orientation of actin filaments near probes due to their long length and rigidity. PMID- 15779989 TI - Specificity and biomineralization activities of Ti-binding peptide-1 (TBP-1). AB - Numerous peptide aptamers that recognize inorganic materials have been isolated using in vitro peptide evolution systems. However, it remains unknown how peptides interact with inorganic materials or how specific those interactions are. We, therefore, assessed the target specificities of the peptide aptamer TBP 1 (RKLPDAPGMHTW) by monitoring its ability to bind 10 different metals. We found that phages displaying TBP-1 bound to Ti, Si, and Ag surfaces but not to Au, Cr, Pt, Sn, Zn, Cu, or Fe. As previously seen with Ti, binding to Si and Ag was diminished by R1A, P4A, or D5A mutation, suggesting that the same molecular mechanism underlies TBP-1 binding to all three materials. We also observed that a synthetic TBP-1 peptide mediated mineralization of both silica and Ag. It, thus, appears that although the overall chemical characteristics of Ti, Si, and Ag surfaces are dissimilar, they share a common subnanometric structure that is recognized by TBP-1. PMID- 15779990 TI - DNA-functionalized MFe2O4 (M = Fe, Co, or Mn) nanoparticles and their hybridization to DNA-functionalized surfaces. AB - Magnetic MFe2O4 (M = Fe, Co, or Mn) nanoparticles with uniform diameters in the 4 20 nm range and with excellent material properties, reported previously, can be rendered soluble in water or aqueous buffers using a combination of alkylphosphonate surfactants and other surfactants such as ethoxylated fatty alcohols or phospholipids. Surfactant-modified oligonucleotides can be incorporated into the particles' organic shell. The particles can withstand salt concentrations up to 0.3 M, temperatures up to 90 degrees C, and various operations such as concentration to dryness, column or membrane separations, and electrophoresis. The particles can be selectively hybridized to DNA functionalized gold surfaces with high coverages using a two-story monolayer structure. These particles may find valuable applications involving the magnetic detection of small numbers of biomolecules using spin valves, magnetic tunnel junctions, or other sensors. PMID- 15779991 TI - Metal-ion adsorption on carboxyl-bearing self-assembled monolayers covalently bound to magnetic nanoparticles. AB - Hematite nanoparticles have been functionalized with a self-assembled monolayer of undecanoic acid (UA) by covalent attachment via siloxane groups. The number density of carboxylic/carboxylate groups on the surface was determined by titration, yielding values on the order of 10(14) UA molecules/cm(2) and thus consistent with a closed-packed monolayer. The ability of these functionalized particles to adsorb Cd(2+) in the ppm range was demonstrated using in situ anodic stripping voltammetry. PMID- 15779992 TI - Property and morphology development in nanocomposite thermoplastic elastomer gels. AB - Thermoplastic elastomer gels (TPEGs), molecular networks composed of a microphase separated multiblock copolymer swollen to a large extent by a low-volatility midblock-selective solvent, are ubiquitous in a wide range of contemporary technologies, including home and office products, athletic equipment, and telecommunications devices. In this work, we investigate the effect of several network-forming nanoscale modifiers-two different silica nanoparticles, three different nanoclays, and a multiwalled carbon nanotube-on the property and morphology development of a TPEG prepared from a microphase-ordered poly(styrene b-(ethylene-co-butylene)-b-styrene) (SEBS) triblock copolymer imbibed with an EB compatible aliphatic mineral oil. Dynamic rheological measurements of the resultant nanocomposite TPEGs confirm that addition of these modifiers affects the linear viscoelastic threshold and increases, to different extents, the dynamic elastic modulus, the dynamic yield stress, and the maximum operating temperature of the parent TPEG. X-ray diffraction analysis reveals that the nanoclays used to generate three series of modified TPEGs are generally swollen with copolymer and/or solvent. PMID- 15779993 TI - Fusogenic tilted peptides induce nanoscale holes in supported phosphatidylcholine bilayers. AB - Tilted peptides are known to insert in lipid bilayers with an oblique orientation, thereby destabilizing membranes and facilitating membrane fusion processes. Here, we report the first direct visualization of the interaction of tilted peptides with lipid membranes using in situ atomic force microscopy (AFM) imaging. Phase-separated supported dioleoylphosphatidylcholine/dipalmitoylphosphatidylcholine (DOPC/DPPC) bilayers were prepared by fusion of small unilamellar vesicles and imaged in buffer solution, in the absence and in the presence of the simian immunodeficiency virus (SIV) peptide. The SIV peptide was shown to induce the rapid appearance of nanometer scale bilayer holes within the DPPC gel domains, while keeping the domain shape unaltered. We attribute this behavior to a local weakening and destabilization of the DPPC domains due to the oblique insertion of the peptide molecules. These results were directly correlated with the fusogenic activity of the peptide as determined using fluorescently labeled DOPC/DPPC liposomes. By contrast, the nontilted ApoE peptide did not promote liposome fusion and did not induce bilayer holes but caused slight erosion of the DPPC domains. In conclusion, this work provides the first direct evidence for the production of stable, well-defined nanoholes in lipid bilayer domains by the SIV peptide, a behavior that we have shown to be specifically related to the tilted character of the peptide. A molecular mechanism underlying spontaneous insertion of the SIV peptide within lipid bilayers and the subsequent removal of bilayer patches is proposed, and its relevance to membrane fusion processes is discussed. PMID- 15779994 TI - AFM characterization of dendrimer-stabilized platinum nanoparticles. AB - This work describes the use of atomic force microscopy (AFM) to measure the size of dendrimer-stabilized Pt nanoparticles (Pt DNs) deposited from aqueous solutions onto mica surfaces. Despite considerable previous work in this area, we do not fully understand the mechanisms by which PAMAM dendrimers template the formation of Pt DNs. In particular, Pt DN sizes measured by high-resolution transmission electron microscopy (HRTEM) are reported to be larger than expected if one assumes that each PAMAM molecule templates one spherical Pt nanoparticle. AFM provides a vertical height measurement that complements the lateral dimension measurement from HRTEM. We show that AFM height measurements can distinguish between "empty" PAMAM and Pt DNs. If the complexation of Pt precursor with PAMAM is prematurely terminated, AFM images and feature height distributions show evidence of arrested precipitation of Pt colloids. In contrast, sufficient Pt PAMAM complexation time leads to AFM images and height distributions that have relatively narrow, normal distributions with mean values that increase with the nominal Pt:PAMAM ratio. The surface density of features in AFM images suggest that these Pt DNs reside on the mica surface as two-dimensional surface aggregates. These observations are consistent with an intradendrimer templating mechanism for Pt DNs. However, we cannot determine if the mechanism obeys a fixed loading law because we do not have definitive information about Pt DN shape. A second peak in the Pt DN height distribution appears when the Pt loading exceeds about 66% of PAMAM's theoretical capacity for Pt. Excluding these secondary particles, the dependence of mean feature height on the Pt:PAMAM ratio follows a power-law relationship. Also considering the magnitudes of the measured mean height values, the data suggest that Pt DNs exist as ramified, noncompact aggregates of Pt atoms interspersed within the PAMAM framework. PMID- 15779995 TI - Patterning polymerized lipid vesicles with soft lithography. AB - The applications of soft lithography in patterning polymerized lipid vesicles of 1,2-bis(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine on glass substrates are reported. We demonstrate that the polymerized vesicles can be used as a high molecular weight ink to be transferred from a PDMS stamp onto a glass substrate to form two-dimensional stripes with a controlled separation. By combining channel flow with dewetting within microfluidic networks, we assemble the polymerized vesicle into three-dimensional stripes and one-dimension lines on glass substrates. Atomic force microscopy shows that these patterned vesicle structures are stable on glass substrates. The simple, stable, and precise immobilization of lipid vesicles on solid substrates will open up the possibility of integrating them in biosensors and microelectronic devices. PMID- 15779996 TI - Fibrous crystalline hydrogels formed from polymers possessing a linear poly(ethyleneimine) backbone. AB - Novel thermoreversible physical hydrogels formed from polymers with linear and star architectures possessing a linear poly(ethyleneimine) (PEI) backbone have been investigated. The hydrogelation occurred simply upon natural cooling of hot aqueous solutions of PEIs to room temperature. The X-ray diffraction and differential scanning calorimetry measurements for the resultant hydrogels unambiguously indicated that the hydrogelation originated from the formation of dihydrate crystalline structures of PEI. These crystalline hydrogels are structurally unique and hierarchical. Microscopic images revealed that the morphologies of the crystalline hydrogels depend on their molecular architectures. The linear PEI resulted in branched fibrous bundles organized by unit crystalline nanofibers with a width of ca. 5-7 nm. The six-armed star with benzene ring core produced fanlike fibrous bundles while the four-armed star with porphyrin core assembled into asterlike aggregates. The critical concentration of gelation (C(G)) was low (about 0.2 approximately 0.3%) and the thermoreversible gel-sol transition temperatures (T(G)) were controllable from approximately 43 to approximately 79 degrees C. The hydrogels formed in the presence of the various aqueous additives including organic solvents, hydrophilic polymers, physical cross-linker, chemical cross-linker, and base enabling modification and functionalization during synthesis. The mechanical properties of the hydrogels could be improved by chemical cross-linking of preformed hydrogels by glutaraldehyde. Physically and physical/chemical cross-linked hydrogels served as excellent template roles in biomimetic silicification, which produced silica-PEI hybrid powder or monolith constructed by nanofibers. PMID- 15779997 TI - Nanomechanical properties of silica-coated multiwall carbon nanotubes-poly(methyl methacrylate) composites. AB - The mechanical properties of polymer composites, reinforced with silica-coated multiwall carbon nanotubes (MWNTs), have been studied using the nanoindentation technique. The hardness and the Young's modulus have been found to increase strongly with the increasing content of these nanotubes in the polymer matrix. Similar experiments conducted on thin films containing MWNTs, but without a silica shell, revealed that the presence of these nanotubes does not affect the nanomechanical properties of the composites. While carbon nanotubes (CNTs) have a very high tensile strength due to the nanotube stiffness, composites fabricated with CNTs may exhibit inferior toughness. The silica shell on the surface of a nanotube enhances its stiffness and rigidity. Our composites, at 4 wt % of the silica-coated MWNTs, display a maximum hardness of 120 +/- 20 MPa, and a Young's modulus of 9 +/- 1 GPa. These are respectively 2 and 3 times higher than those for the polymeric matrix. Here, we describe a method for the silica coating of MWNTs. This is a simple and efficient technique, adaptable to large-scale production, and might lead to new advanced polymer based materials, with very high axial and bending strength. PMID- 15779998 TI - Two-dimensional ordered arrays of aligned lipid tubules on substrates with microfluidic networks. AB - Microfluidic networks is a powerful tool for aligning one-dimensional materials over a large area on solid substrates. Here we show that lipid nano- and microtubules can be assembled into two-dimensional (2-D) parallel arrays with controlled separations by combining fluidic alignment with dewetting, which occurs within microchannels. We also demonstrate that lipid tubules can be bent into a well-defined shape at the entrance of the channels by the capillary force. Atomic force microscopy is used to study the structure and stability of the aligned lipid tubules on substrates. The deposition experiments with silica colloidal particles show that the 2-D parallel-aligned tubules can be used as a template to synthesize silica films with controlled morphologies and patterns on substrates in a single-step process. PMID- 15779999 TI - Inward-growing self-assembly of colloidal crystal films on horizontal substrates. AB - Colloidal crystal films have been fabricated on solid substrates with a horizontal deposition method. Scanning electron microscope images showed that the colloidal crystal films exhibit ordered face-centered cubic structures in large domains. Optical measurements demonstrated the presence of photonic band gap along the crystallographic [111] direction. The fabrication method described in this paper allows one to rapidly fabricate colloidal crystal films of different thicknesses, which can be controlled by varying colloidal suspension concentration or volume. In addition, the method also works well for growing colloidal crystal films on a hydrophilic solid substrate with a rough surface. Furthermore, the fabrication of colloidal crystal heterostructures has been demonstrated. An inward-growing mechanism responsible for self-assembly of colloidal spheres on horizontal substrates has been proposed to interpret the observed experimental results. PMID- 15780000 TI - Salt softening of polyelectrolyte multilayer capsules. AB - The changes in the morphology and the mechanical properties of hollow polyelectrolyte multilayer capsules made from poly(styrenesulfonate)/poly(allylamine hydrochloride) in response to added salt were investigated. We found that capsules shrink in response to salt exposure. The effect depends strongly on the nature of the salt added and follows trends of the Hoffmeister series, with weakly hydrated cations inducing the strongest shrinking. For NaCl, we have investigated additional effects on capsule mechanical properties that are occurring above a 3 M salt concentration and we found that the morphological changes are accompanied by a pronounced softening of the capsule wall material, which we can quantify by analyzing the force response of capsules in the prebuckling regime. This shows that salts can act as plasticizers in the multilayers and induce annealing effects. PMID- 15780001 TI - Characterization of TiO2 nanoparticles surface modified with aluminum stearate. AB - This paper uses measurements of adsorption and vibrational spectra (DRIFTS, ATR, and Raman) to characterize TiO2 (rutile) nanoparticles that have been surface treated with aluminum and stearate, "aluminum stearate". From these measurements, we have developed a model of titania particles covered by patches of "alumina". Vibrational spectra, particularly the spectra of the carboxylate headgroups, show that the stearate then adsorbs on both the titania and the alumina. Surprisingly, the distribution of the stearate between alumina and titania is sensitive to the presence of water. As the water content decreases, the relative amounts of stearate on titania, rather than alumina, increase, and this increase is accompanied by a less ordered structuring of the stearate tails, as evidenced by a shift of the C-H stretching bands to higher frequencies and a broadening of the 1296 cm(-1) Raman band. This effect is consistent with earlier observations that the presence of water reduced the bonding of stearate headgroups to the surface of titania. We have also shown that the dispersion in C12-C15 alkyl benzoate of aluminum stearate coated titania is sensitive to the presence of small amounts, approximately 4%, of water. Finally, we have demonstrated that surface stearate, like surface alumina, reduces the rate of phototocatalytic oxidation of 2 propanol. A 7% stearate coating reduces acetone formation by a factor of 4. There is no evidence from these studies that, during the oxidation experiment, 2 propanol displaces stearate from the titania surface. PMID- 15780002 TI - Dynamic aspects of mercury porosimetry: a lattice model study. AB - Grand canonical Monte Carlo simulations using both Glauber dynamics and Kawasaki dynamics have been carried out for a recently developed lattice model of a nonwetting fluid confined in a porous material. The calculations are aimed at investigating the molecular scale mechanisms leading to mercury retention encountered during mercury porosimetry experiments. We first describe a set of simulations on slit and ink-bottle pores. We have studied the influence of the pore width parameter on the intrusion/extrusion curve shapes and investigated the corresponding mechanisms. Entrapment appears during Kawasaki dynamics simulations of extrusion performed on ink-bottle pores when the system is studied for short relaxation times. We then consider the more realistic and complex case of a Vycor glass building on recent work on the dynamics of adsorption of wetting fluids (Woo, H. J.; Monson, P. A. Phys. Rev. E 2003, 67, 041207). Our results suggest that mercury entrapment is caused by a decrease in the rate of mass transfer associated with the fragmentation of the liquid during extrusion. PMID- 15780003 TI - Prediction of high-pressure adsorption equilibrium of supercritical gases using density functional theory. AB - In this paper, we present the results of the prediction of the high-pressure adsorption equilibrium of supercritical gases (Ar, N2, CH4, and CO2) on various activated carbons (BPL, PCB, and Norit R1 extra) at various temperatures using a density-functional-theory-based finite wall thickness (FWT) model. Pore size distribution results of the carbons are taken from our recent previous work,(1,2) using this approach for characterization. To validate the model, isotherms calculated from the density functional theory (DFT) approach are comprehensively verified against those determined by grand canonical Monte Carlo (GCMC) simulation, before the theoretical adsorption isotherms of these investigated carbons calculated by the model are compared with the experimental adsorption measurements of the carbons. We illustrate the accuracy and consistency of the FWT model for the prediction of adsorption isotherms of the all investigated gases. The pore network connectivity problem occurring in the examined carbons is also discussed, and on the basis of the success of the predictions assuming a similar pore size distribution for accessible and inaccessible regions, it is suggested that this is largely related to the disordered nature of the carbon. PMID- 15780004 TI - On the mechanism of selective electroadsorption of protons in the pores of carbon molecular sieves. PMID- 15780006 TI - Characterization of a spontaneously polarizing HT-29 cell line, HT-29/cl.f8. AB - A cloned cell line that spontaneously polarizes in standard glucose-containing media was derived from a single cell of the adenocarcinoma cell line HT-29. The cloned line, designated HT-29/cl.f8, has remained stable over 2 yr in culture, maintained high transepithelial resistance (300 ohm cm(2) or higher), and correctly sorted influenza virus and vesicular stomatitis virus to apical or basolateral domains, respectively. The newly cloned cells also displayed apical microvilli, tight junctions, and desmosomes, the morphological characteristics of mature epithelia. The cloned HT-29/cl.f8 cells function as epithelial enterocytes as shown by the apical expression of intestinal alkaline phosphatase, the expression of vimentin and cytokeratin, and lack of expression of mucin. We propose that the newly cloned HT-29/cl.f8 cells offer a viable alternative for studies of enterocyte function that will readily yield interpretable data not complicated by cell alterations due to the presence of drugs or chemicals that induce differentiation. PMID- 15780008 TI - Characterization of cultured insect cells selected by Bacillus thuringiensis crystal toxin. AB - Selection for resistance against Bacillus thuringiensis (Bt) Cry1Ac10 in the Trichoplusia ni (Hubner) cell line BTI-TN-5B1-4 (TnH5) was tested, and the development of resistance in the selected cells was like a S-form curve. Monitoring at the Cry1Ac10 50th challenge, the resistance ratio was 1,294-fold as many as that of initial cells. But the resistance to Cry1Ac10 declined gradually when the selection was relaxed. The resistance declined rapidly at the low level of resistance and slowly at the high level of resistance. This resistant cell had high resistance to all the tested solubilized trypsin-treated mixture of crystal multitoxins of B. thuringiensis subsp. aizawai GC-91, an engineering bacterium of Bt, B. thuringiensis subsp. aizawai HD-133 and B. thuringiensis subsp. kurstaki HD-1, and low cross-resistance (19.7-fold) to activated Cry1C. Both N-acetyl-D galactosamine (GalNAc) and tunicamycin did not inhibit the toxicity of Cry1Ac10 to the susceptible TnH5 cells. Comparison of the total proteins of the selected resistant cells with that of the nonselected susceptible cells by two-dimensional electrophoresis analysis showed that were obvious differences among the 11 protein expressions. These results strongly suggest that there exists an unknown mechanism of resistance in the cell line that was different from the reported mechanisms in insects. PMID- 15780005 TI - The Wolbachia genome of Brugia malayi: endosymbiont evolution within a human pathogenic nematode. AB - Complete genome DNA sequence and analysis is presented for Wolbachia, the obligate alpha-proteobacterial endosymbiont required for fertility and survival of the human filarial parasitic nematode Brugia malayi. Although, quantitatively, the genome is even more degraded than those of closely related Rickettsia species, Wolbachia has retained more intact metabolic pathways. The ability to provide riboflavin, flavin adenine dinucleotide, heme, and nucleotides is likely to be Wolbachia's principal contribution to the mutualistic relationship, whereas the host nematode likely supplies amino acids required for Wolbachia growth. Genome comparison of the Wolbachia endosymbiont of B. malayi (wBm) with the Wolbachia endosymbiont of Drosophila melanogaster (wMel) shows that they share similar metabolic trends, although their genomes show a high degree of genome shuffling. In contrast to wMel, wBm contains no prophage and has a reduced level of repeated DNA. Both Wolbachia have lost a considerable number of membrane biogenesis genes that apparently make them unable to synthesize lipid A, the usual component of proteobacterial membranes. However, differences in their peptidoglycan structures may reflect the mutualistic lifestyle of wBm in contrast to the parasitic lifestyle of wMel. The smaller genome size of wBm, relative to wMel, may reflect the loss of genes required for infecting host cells and avoiding host defense systems. Analysis of this first sequenced endosymbiont genome from a filarial nematode provides insight into endosymbiont evolution and additionally provides new potential targets for elimination of cutaneous and lymphatic human filarial disease. PMID- 15780009 TI - Liver-specific physiology of immortal, functionally differentiated hepatocytes and of deficient hepatocyte-like variants. AB - Five different immortalized transgenic hepatocyte cell lines derived from mice were investigated with respect to their potential to maintain the physiological properties of primary hepatocytes using chemically defined medium. This research completes a previous study by Klocke and coworkers in 2002, using gene expression analysis of the same cell lines by the respective physiological analysis for investigating the hepatocyte-like function. Three transgenic cell lines harboring a fusion gene derivative (construct 202) consisting of the complete SV40 early region, including the coding sequences for the transforming large and small tumor antigens, placed under the control of the murine metallothioneine 1 promotor/enhancer element, showed a hepatocyte-like function and physiology. They grew as a monolayer with a polygonal cell shape, consumed lactate, and secreted albumin at a cell-specific rate of 1.5 pg/h, which is in the range of primary hepatocytes. In addition, the potential of detoxifying ammonium could be maintained. Ammonium was metabolized and urea was produced and released into the medium. A complete urea cycle could be determined. A cell line established from neonatal transgenic mice and expressing a secretory variant of the human epidermal growth factor (IgEGF) under the control of the albumin promoter was characterized by an incomplete urea cycle. Another cell line isolated from the liver of homozygote neonatal p53-knockout mice showed no hepatocyte-specific functions but only properties of continuous cell lines. Specific nucleoside triphosphate (NTP) and uridine (U) ratios were used to characterize the differentiation status of the particular cell lines. A low NTP-U value was found for the three cell lines containing construct 202, which was identical to that observed for primary hepatocytes. In contrast, the cell line harvested from the liver of homozygote neonatal p53-knockout mice presented a NTP-U ratio characteristic for continuous cell lines. This study demonstrates that the four transgenic and the p53-knockout hepatocyte-derived cell lines can be used as models for investigating the conservation of tissue-specific functions in immortalized cells. PMID- 15780007 TI - Differentiated cultures of primary hamster tracheal airway epithelial cells. AB - Primary airway epithelial cell cultures can provide a faithful representation of the in vivo airway while allowing for a controlled nutrient source and isolation from other tissues or immune cells. The methods used have significant differences based on tissue source, cell isolation, culture conditions, and assessment of culture purity. We modified and optimized a method for generating tracheal epithelial cultures from Syrian golden hamsters and characterized the cultures for cell composition and function. Soon after initial plating, the epithelial cells reached a high transepithelial resistance and formed tight junctions. The cells differentiated into a heterogeneous, multicellular culture containing ciliated, secretory, and basal cells after culture at an air-liquid interface (ALI). The secretory cell populations initially consisted of MUC5AC-positive goblet cells and MUC5AC/CCSP double-positive cells, but the makeup changed to predominantly Clara cell secretory protein (CCSP)-positive Clara cells after 14 d. The ciliated cell populations differentiated rapidly after ALI, as judged by the appearance of beta tubulin IV-positive cells. The cultures produced mucus, CCSP, and trypsin-like proteases and were capable of wound repair as judged by increased expression of matrilysin. Our method provides an efficient, high-yield protocol for producing differentiated hamster tracheal epithelial cells that can be used for a variety of in vitro studies including tracheal cell differentiation, airway disease mechanisms, and pathogen-host interactions. PMID- 15780012 TI - Overweight status of adolescent girls with polycystic ovary syndrome: body mass index as mediator of quality of life. AB - PURPOSE: Adolescent girls with polycystic ovary syndrome (PCOS) have significant disruption in health-related quality of life (HRQL) compared to their healthy peers, but it is unclear which aspects of the disorder have the greatest impact on these quality-of-life changes. Studies of adult women have indicated that weight status has a significant impact on subjective symptomatology; however, a similar effect of weight status on HRQL in adolescents with PCOS has not been established. This study evaluated the effect of body mass index (BMI) on quality of-life disturbances in adolescent girls with PCOS. METHODS: Data were derived from the Adolescent PCOS Quality of Life Study, a cross-sectional study of healthy (n = 186) and PCOS-affected adolescent girls (n = 97). The study was conducted at an urban, hospital-based adolescent medicine clinical practice. Medical staff obtained anthropomorphic measurements. Participants in both groups completed the Child Health Questionnaire-Version CF-87 as a measure of HRQL. BMI was tested as a mediator of HRQL using bivariate analyses and multivariate linear regression models. RESULTS: The average BMI was 31.7 kg/m(2) (standard deviation [SD] = 8.4) for adolescents with PCOS and 23.5 kg/m(2) (SD = 4.2) for healthy adolescents. BMI was associated with PCOS status and HRQL. Girls with PCOS scored lower on the general health perceptions scale, physical functioning scale, family activities scale, and the general behavior scale and higher on the change in health in the last year subscale. When BMI was added to the multivariate linear regression models, coefficients were reduced on average by 3 points and became nonsignificant, indicating that the HRQL differences are mediated by the higher BMI. CONCLUSION: This study demonstrates that BMI is a primary mediator in the relationship between PCOS and the HRQL reductions experienced by girls with the disorder. Clinical interventions that effectively address weight issues in adolescents with PCOS may improve overall HRQL and other obesity-related clinical outcomes. PMID- 15780011 TI - Emergency department utilization by children in foster care. AB - OBJECTIVE: To examine emergency department (ED) utilization of children in foster care using nationally representative data. METHODS: Cross-sectional analysis of the National Survey of Child and Adolescent Well-Being, Wave 1, which provided data for children ages 1 to 14 years in foster care for 1 year between October 1999 and December 2000. We identified children as having used the ED based upon foster parent responses to the question, "In the last 12 months, has your child gone to an emergency room or urgent care center (UCC) for an illness or injury?" We examined child and family demographic variables that were associated with having used the ED. RESULTS: The sample included 559 children in foster care. Thirty-one percent (95% confidence interval = 26%, 36%) of foster children had visited the ED or UCC in the past 12 months. Multivariate logistic regression analysis revealed that out of all children in foster care, children who had a chronic condition, children of younger age, and children with younger foster caregivers were significantly more likely to have used the ED. CONCLUSION: A sizeable proportion of children in foster care use the ED, and subgroups of these children have particularly high rates of ED use. An important component of providing a medical home for children in foster care should involve coordination of care around ED visits for this vulnerable group of children. PMID- 15780010 TI - Interleukin-8 can stimulate progesterone secretion from a human trophoblast cell line, BeWo. AB - Precise paracrine cross-talk between the embryo and the endometrium is essential for the establishment of a successful pregnancy. Previous studies have demonstrated that the expression of interleukin-8 (IL-8) in the endometrium is enhanced during the late-secretory phase and early pregnancy. Furthermore, IL-8 receptor (IL-8R) expression has been detected in trophoblast cells of the developing embryo. To clarify the roles of IL-8 in the endometrium-embryo interactions, the effects of IL-8 on hormone secretion by trophoblast cells were studied using the BeWo trophoblast cell line that retains hormone-secreting properties of normal trophoblast cells. Using reverse transcription-polymerase chain reaction, we found that IL-8R messenger ribonucleic acid (mRNA) was expressed in BeWo cells. The levels of IL-8R mRNA and protein expression in BeWo cells were similar to those in primary first-trimester trophoblast cells. Progesterone (P4) secretion of BeWo cells was comparable with that of first trimester trophoblast cells but higher than that of third-trimester trophoblast cells. Treatment of BeWo cells with recombinant human IL-8 (rhIL-8) had no effect on cell proliferation, as determined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide assay. Interestingly, secretion of P4, but not human chorionic gonadotropin, from cultured BeWo cells was significantly enhanced when the cells were incubated with rhIL-8. Our results demonstrate that IL-8 may play an important role in the endometrium-embryo interactions by stimulating trophoblast secretion of P4 for maintenance of a successful pregnancy. PMID- 15780013 TI - Precepting genital exams: challenges in implementing a complex educational intervention in the continuity clinic. AB - BACKGROUND: The female genital exam is often omitted from the routine physical. This limits familiarity with normal findings and represents a missed opportunity to evaluate girls for urogenital abnormalities. Continuity clinic offers an opportunity to develop and test a teaching intervention to address this problem. OBJECTIVE: To describe the effect of precepted genital exams on resident confidence and comfort with the exam as well as knowledge and documentation of genital anatomy. METHODS: A controlled trial was conducted in 10 continuity clinics at Children's Hospital, Columbus, Ohio. Residents in 5 clinics reviewed genital anatomy, exam positions, and documentation examples. They were asked to complete 6 precepted exams of girls aged 1-6 years during well-child care over a 6-month period. Residents were given pre- and posttests assessing knowledge of, confidence, and comfort with the genital exam. Pre- and poststudy genital exam documentation was reviewed. RESULTS: Ninety-seven percent of the intervention group, compared to 73% of the control group, answered 4 or 5 on a 5-point scale of confidence with regard to recognizing a normal exam (5 = highest) following the intervention (P = .01). Ninety-four percent of the intervention group and 64% of the control group answered 4 or 5 with regard to how comfortable they were performing genital exams (P = .01). Knowledge scores and documentation of genital anatomy did not differ between groups. Thirty-two percent of residents had the full intervention. CONCLUSIONS: Resident confidence and comfort increase with precepting. Precepting did not improve knowledge or documentation of genital anatomy. We advocate use of this preliminary data to design and test future educational interventions. PMID- 15780014 TI - Internet and computer access and use for health information in an underserved community. AB - BACKGROUND: The "digital divide" is the gap between those with access to information tools such as the Internet and those without access. The gap has been described by income, education, age, and race. Little information exists on computer and Internet access and use for health information by parents, particularly among populations of low income and low education level. OBJECTIVE: To describe computer and Internet access and use, including health information retrieval, among low-income, urban, African American caregivers (parents). DESIGN: Cross-sectional survey administered in pediatric waiting rooms of urban community-based health centers in a low-income area. PARTICIPANTS: Caregivers of pediatric outpatients. MAIN OUTCOME MEASURES: Access to computers, Internet access, and use of Internet for health information. RESULTS: In 2003, among 260 African Americans who completed surveys, 58% had a computer and 41% had home Internet access. Fifty-two percent had used the Internet for finding health information. Ninety-three percent agreed or strongly agreed that there is useful health information on the Internet. Ninety-two percent agreed or strongly agreed that they would want to talk with a medical professional about health information on the Internet. Sixty-five percent of respondents had no additional schooling after high school. Annual household income was 0.05). Therefore, hepatitis B infection might be a contributing factor for cholangiocarcinoma development. PMID- 15780028 TI - Peer education project on breast self-examination in Izmir, Turkey. AB - PURPOSE: The purpose of this research was to assess undergraduate female student knowledge related to the Peer Education Method and Breast Self-Examination (BSE), to encourage them to refer for screening with certain intervals and to teach them the means of utilizing the health services. DESIGN: The subjects comprised 160 undergraduate female students who were selected by simple random sampling and who accepted to participate. METHODS: A survey form, which was prepared under the direction of relative literature by the researcher four 3rd graders of Ege University School of Nursing, was applied to female students. The data collected were assessed after being coded. FINDINGS: The numbers of undergraduate female students performing BSE regularly and their information levels related to the symptoms of breast cancer were found to be very low, but their mean scores for performing BSE after training given by peer guides were increased and the peer group training was found to be effective. CONCLUSIONS: From the results we conclude that the training guide was appropriate for transmitting information to university students for peer group education purposes. PMID- 15780029 TI - A case-control study of multiple myeloma in Japan: association with occupational factors. AB - Recently the incidence rate of multiple myeloma (MM) has increased in Japan. Epidemiologic efforts have suggested that certain occupational and chemical exposures are likely to increase the risk for MM. We therefore performed a case control study of MM, examining occupational factors. Data for 57 cases and 57 controls were obtained from Sapporo Medical University Hospital and its affiliated hospitals in Hokkaido. Controls were matched to each case by gender, age (+/- 5 years) and hospital. Detailed information regarding several factors possibly related to MM was obtained by interviews in hospitals. Odds ratios (ORs) and 95% confidence intervals (95%CIs) were calculated with adjustment for cigarette smoking as confounding factor. The occupational category of agriculture and fishery showed a significant association with increased risk (OR = 5.89, 95% CI = 1.24-28.04). Occupational exposure to chemical products including organic solvents or petroleum showed a significant association with increased risk (OR = 8.05, 95% CI = 1.01-64.45). Medical histories of operation for appendicitis and blood transfusion were associated with decreased risk, but not significantly. Our results suggested that occupational exposure to chemicals might play a role in the risk of MM. PMID- 15780030 TI - Epidemiology of breast cancer in females of reproductive age in Kyrgyzstan. AB - The goal of this research was to study breast cancer morbidity in females of reproductive age in Kyrgyzstan. Information on patients was obtained from the National Center of Oncology under the Ministry of Health and the National Statistics Committee of the Kyrgyz Republic. The research was retrospective and covered the period from 1995-2002. Cancer morbidity ratios were calculated for reproductive age according to standard methods of medico-biological statistics. The breast cancer morbidity in the country's female population was determined as 12.3+/- 0.2/100,000. The research revealed ethnic specificity: in Russians (crude rate, 32.9+/- 2.1) was higher (p<0.001), than in Kyrgyz and Uzbek females, who demonstrated equal crude incidence rates of -8.0 +/- 0.6. The dynamics over time showed increase in Kyrgyz and Uzbek females but decrease in Russians. Age ratios analysis showed higher morbidity in later reproductive age (40-49 years), with a statistically significant difference (p<0.001) between ethnic Europeans and Asians. PMID- 15780031 TI - Dietary administration of inositol and/or inositol-6-phosphate prevents chemically-induced rat hepatocarcinogenesis. AB - Chemoprevention is considered a rational strategy for dietary approaches to prevention of cancer. Multiple lines of evidence suggest that many of our dietary principles are able to intervene in the multistage carcinogenesis process and phytic acid (inositol hexaphosphate, IP6), a phytochemical present in a variety of plant species, has been shown to prevent various cancers, including those of the mammary gland, colon and liver. However, the mechanism of chemoprevention by IP6 has not been fully elucidated. In the present study, we examined the effects of inositol and/or IP6 supplementation on rat hepatocarcinogenesis initiated by diethylnitrosamine (DEN) and promoted by partial hepatectomy (PH). Supplementation with either inositol or IP6, or their combination, starting one week prior to administration of DEN, resulted in a significant decrease in both the area and the number of placental glutathione S-transferase positive (GST-P+) foci, a preneoplastic marker for DEN-initiated hepatocarcinogenesis. The administration of inositol and/or IP6 in drinking water caused marked enhancement in the glutathione S-transferase (GST) activity. In addition, the production of thiobarbituric acid reactive substances and the catalase activity were significantly reduced in rats supplemented with inositol and /or IP6. Based on these findings, it is likely that the chemopreventive effects of inositol and/or IP6 on rat hepatocarcinogenesis initiated by DEN and promoted by PH are associated with induction of GST activity and suppression of lipid peroxidation. PMID- 15780032 TI - Risk factors for esophageal cancer: a case-control study in South-western China. AB - Esophageal cancer is a crucial cancer in China. Yanting in Sichuan Province was a key area with highest esophageal cancer mortality in China, but little evidence on esophageal cancer risk factors has been reported for this area and the etiology remains unclear. To clarify risk factors, a 1:1 matched case-control study was conducted. Totals of 185 eligible esophageal cancer patients and 185 healthy residents matched for sex and age were recruited. Conditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for possible risk/protective factors. All ORs were adjusted by family history of esophageal cancer and occupation, and then further adjusted by other possible confounding factors. Our results showed that smoking and alcohol drinking were risk factors for esophageal cancer with dose-response. The ORs (95% CI) compared with never smokers and drinkers were 4.06 (1.55-10.6) and 2.49 (1.06 5.85), respectively. The OR was further increased to 8.86 (95% CI, 3.82-20.5) for both smoking and drinking in combination. Eating food rapidly (OR=5.84, 95% CI, 2.05-16.7), drinking shallow ground water (OR=4.18, 95% CI, 1.30-13.4) and frequent intake of picked vegetables (OR=2.12, 95% CI, 1.00-4.49) appeared to increase the risk, while frequent intake of fresh fruit (OR=0.42, 95% CI, 0.19 0.89), fresh vegetables (OR= 0.62, 95% CI, 0.32-1.17) and eggs (OR=0.59, 95% CI, 0.25-1.39) decreased the risk. In conclusion, smoking and alcohol drinking are common in Yanting and main contributors to esophageal cancer. Consumption of fresh fruit and eggs are not common and high consumption of these two foods as well as fresh vegetables may decrease the risk of esophageal cancer in this area. In addition, drinking shallow ground water and eating food rapidly, as well as frequent intake of pickled vegetables, are also factors increasing the risk. PMID- 15780033 TI - Variables associated with recurrence in breast cancer patients-the Shaukat Khanum Memorial experience. AB - From a cohort of female breast cancer patients registered at the Shaukat Khanum Memorial Cancer Hospital and Research Center, in Lahore, Pakistan, during the time period extending from December 1994 to December 2002, 700 subjects who were followed up in time, were selected. Those who presented with benign tumors, carcinoma in situ, or metastases were excluded from the analyses. Age, tumor size, nodal status, menopause, estrogen receptor (ER), and progesterone receptor (PR) status, at the time of presentation, were determined. Tumors were classified according to the TNM classification (American Joint Commission on Cancer (AJCC) sixth edition), and subsequently, grouped into T1/T2 and T3/T4. Lymph nodes were categorized as N0 (node-negative) and N1, N2, and N3 combined (node-positive). The odds ratio (OR) for developing recurrence in T3/T4 versus T1/T2 was determined to be 2.06 (95% confidence interval (CI) 1.39-3.05, p < 0.001); the OR for node-positive relative to node-negative was found to be 2.54 (95 % CI 1.61 4.0, p < 0.001). Furthermore, the association between the odds of developing recurrence in ER-positive compared to ER-negative was represented by an OR of 0.61, (95 % CI 0.40-0.94 (p= 0.02)). These findings are consistent with the observations that ER-positive, node-negative, and T1/T2 lesions have a decreased risk of recurrence. Also, ER-positive patients may have a better response to hormonal treatment than those who are ER-negative. PMID- 15780034 TI - Awareness of oral cancer and precancer among patients attending a hospital in Sri Lanka. AB - OBJECTIVE: To assess awareness about oral cancer and precancer among patients attending for dental treatment at a University Dental hospital. METHODS AND MATERIALS: A self-administered questionnaire was used to collect information from 410 randomly selected outpatients attended the Dental Hospital (teaching), Faculty of Dental Sciences, University of Peradeniya, Sri Lanka. The questionnaire included questions to ascertain information on socio-demographic parameters, awareness of oral cancer and precancer, habits of betel chewing, smoking and alcohol consumption. RESULTS: Ninety five percent of the respondents were aware of the possibility of occurrence of cancer in the mouth while only 44.9% (n=184) were aware about precancer. Of the 390 individuals who were aware of the existence of oral cancer, 80.7% were knowledgeable about the causal relationship between betel chewing habit and oral cancer. Forty-seven and 17 percent were aware of links with tobacco smoking and alcohol consumption, respectively. However, out of those who had knowledge of oral cancer, 18 % (n=69) were not knowledgeable about associations with habits. CONCLUSIONS: This survey revealed that the patients attending the hospital were well informed about oral cancer. However, awareness about precancer was relatively low. Knowledge about the causal relationships with tobacco smoking and use of alcohol was low compared to that for betel chewing. PMID- 15780035 TI - Relation of the CD36 gene A52C polymorphism to the risk of colorectal cancer among Japanese, with reference to with the aldehyde dehydrogenase 2 gene Glu487Lys polymorphism and drinking habit. AB - High consumption of white meat (or saturated fatty acids) and alcohol has been demonstrated to have a tendency to increase the risk of colorectal cancer, according to the level of malondialdehyde-deoxyguanosine adducts derived from lipid per-oxidation in the colorectal mucosa. CD36 plays important roles as a long-chain fatty acid translocase and oxidized low-density lipoprotein (LDL) scavenger, while alcohol is metabolized by aldehyde dehydrogenase 2 (ALDH2) and decreases transiently metabolism of dietary fat and serum lipids. To examine associations between the risk of colorectal cancer and the CD36 gene A52C polymorphism according to the ALDH2 gene Glu487Lys polymorphism and drinking habit, a hospital-based case-control study was conducted with 128 colorectal cancer cases and 238 cancer-free controls. Odds ratios (ORs) for the C/C genotype relative to the A/A genotype were 1.70 [95% confidence interval (CI), 0.76-4.11] and 4.24 (95% CI, 1.42-22.66) for men and women, respectively, with the low activity (Glu/Lys + Lys/Lys) ALDH2 genotype. The high-activity (Glu/Glu) genotype for men and women had no associations. On the other hand, the OR for the C/C genotype with high frequency of drinking habit relative to the A/A genotype with low frequency of drinking habit among men was 3.63 (95% CI, 1.29-13.15). The number of women with a high frequency drinking habit was too small for any corresponding analyses. Our findings suggest a significant interaction between alcohol consumption and the CD36 gene A52C polymorphism related to the metabolism of long-chain fatty acids and oxidized LDL in the etiology of colorectal cancer. PMID- 15780036 TI - Accuracy of visual inspection with acetic acid (VIA) for early detection of cervical dysplasia in Tehran, Iran. AB - OBJECTIVE: To evaluate the accuracy of visual inspection with 5% acetic acid (VIA) when used to detect cervical cancer and its precursors. METHODS: The study population included women attended Family Planning and Gynecological Clinic in Bagher Abad Health Center and Mirza Koochak Khan Hospital for regular cervical screening tests. After obtaining informed consent from each woman, VIA was performed. One hundred with a positive VIA test and 100 women with a negative VIA test were randomly selected for this study. Cytology and colposcopy examination were performed for all 200 cases and cervical biopsies were conducted for those individuals showing abnormal colposcopic findings. RESULTS: Nine cases in VIA positive group and two cases in VIA-negative group had an abnormal cytology. Ninety five women in the VIA-positive group and 25 in the VIA-negative group had abnormal colposcopic findings. From biopsy examination, 67 (71%) of cases in the VIA-positive group and 3 (12%) cases in the VIA-negative group had a final diagnosis of dysplasia. Among biopsied samples, only 7 cases of VIA-positive group showed abnormal result and the remaining were normal. Based on these results, VIA test sensitivity and specificity were 95.7% and 44.0% respectively, while they were 10% and 92% for cytology tests. CONCLUSIONS: The results of this study indicate that although VIA is a sensitive screening test for detection of cervical dysplasia, it can not be used by itself. Applying VIA along with Pap smears helps to detect a higher number of cases with cancer precursor lesions. PMID- 15780037 TI - Population based survival of female breast cancer cases in Riyadh Region, Saudi Arabia. AB - Breast cancer is the most frequent tumor among Saudi women, accounting to 19.8% of female cancers. The present study was conducted to determine 5-year survival for all cases of invasive breast cancer that occurred during 1994-96 in the province of Riyadh (n=316). The overall observed survival probability of the study population at 1, 3 and 5 years was 93.9%, 79.2% and 59.6%, respectively. The 5 year survivals for the younger (< 40 years), older (50 + years) and 40-49 years patients were 60.6%, 51.6% and 69.2% respectively, the differences not reaching statistical significance. While there was not a great deal of variation in the 5-year survival between cases with regional (55.6%), distant metastasis (57.6%) and extent of disease unknown (56.7%) cases, localized (67.5%) cases had a clearly better prognosis. An increased but not significant hazard was seen for the cases with regional and distant metastasis disease, 1.40 and 1.11 respectively, compared to localized cases. The 5-year survival for duct carcinomas (62.8%) was greater than for adenocarcinomas (55.6%) and lobular carcinomas (50.0%). PMID- 15780038 TI - Olive: fruit of peace against cancer. AB - The olive has a history almost as long as that of Western civilization and has been looked upon as a sign of hope, peace, and sacredness. Olive oil, extracted from the olive, is the principal source of dietary fat in the Mediterranean basin. The composition differs from that of other dietary fats in that olive oil is rich in monounsaturated fatty acids. Even other than as a source of monounsaturated fats, olive oil has own unique effects. Accumulating evidence suggests that olive oil may have beneficial health effects, especially when it comes to reducing risk factors for coronary heart diseases, preventing cancer, and modifying immune and inflammatory responses. However, evidence remains limited, definitive conclusions are difficult to draw, and there remains a significant need for further studies, particularly prospective cohort and well designed, controlled intervention studies. In this manuscript, the beneficial health effects of olive oil are reviewed, with particular attention paid to cancer prevention and immune functions.. PMID- 15780039 TI - Screening for breast cancer -is there an alternative to mammography? AB - Given the continuing increase in mammary cancer incidence and in many cases also mortality across the world, as well as the difficulty with primary prevention, the question of whether screening for early detection is effective is of prime importance. If there is a real benefit in terms of reduced mortality then attention should clearly be focused on the modality which should be recommended in different resource settings. In the developed world where mammography is generally available the results are less than conclusive. It seems possible that there is a segment of breast cancer benefited both by screening and by treatment, and that far from these effects being additive, they affect the same spectrum of cases, so that as treatment improves, the benefit we can expect to see from screening falls. In the Asian Pacific setting, randomized trials on the basis of the cost and benefit should be a high priority. However, the lesson from all programmes of breast screening, is that for success, attention has to be paid to all aspects of the programme, compliance with screening, high quality screening tests, quality in the referral, diagnosis and treatment process, as well as adequate follow-up. PMID- 15780040 TI - Hepatitis B virus and human immunodeficiency virus co-infection in sub-Saharan Africa: a call for further investigation. AB - A growing body of evidence indicates that human immunodeficiency virus (HIV) positive individuals are more likely to be infected with hepatitis B virus (HBV) than HIV-negative individuals, possibly as a result of shared risk factors. There is also evidence that HIV-positive individuals who are subsequently infected with HBV are more likely to become HBV chronic carriers, have a high HBV replication rate, and remain hepatitis Be antigen positive for a much longer period. In addition, it is evident that immunosuppression brought about by HIV infection may cause reactivation or reinfection in those previously exposed to HBV. Furthermore, HIV infection exacerbates liver disease in HBV co-infected individuals, and there is an even greater risk of liver disease when HIV and HBV co-infected patients are treated with highly active anti-retroviral therapy (HAART). Complicating matters further, there have been several reports linking HIV infection to 'sero-silent' HBV infections, which presents serious problems for diagnosis, prevention, and control. In sub-Saharan Africa, where both HIV and HBV are endemic, little is known about the burden of co-infection and the interaction between these two viruses. This paper reviews studies that have investigated HIV and HBV co-infection in sub-Saharan Africa, against a backdrop of what is currently known about the interactions between these two viruses. PMID- 15780042 TI - Influence of viral load and genotype in the progression of Hepatitis B-associated liver cirrhosis to hepatocellular carcinoma. AB - AIM/BACKGROUND: Hepatitis B virus (HBV) is an important factor in the development of hepatocellular carcinoma (HCC). We studied the influence of HBV viral load on HCC occurrence in HBV related liver cirrhosis (LC). PATIENTS AND METHODS: Ninety one LC patients were followed up over a period of 7 years. Twenty three patients received Interferon (IFN) therapy. RESULTS: In 7 years, 23 patients developed HCC. Of them twenty-two (95.6%) were of genotype C. HBV DNA was found to be the only significant variable associated with HCC occurrence on both univariate (P = 0.029) and multivariate analysis (odds ratio 2.33; P < 0.033). The cumulative survival at 5 years was 83% and the annual rate of hepatitis B surface antigen clearance was 0.9 %. All of 17 HCC patients observed over a period of 5 years or more belonged to the continuously high HBV DNA group (annual average >3.7 log copies/ml) and all but one belonged to the continuously high alanine aminotransferase group (annual average >40 IU/l). CONCLUSION: Patients with genotype C and a continuously high HBV DNA for 5 years or more are at a high-risk group for HCC development. Maintaining continuously low HBV DNA for 3 years or more with anti-viral therapy, may be useful in preventing or delaying HCC occurrence. PMID- 15780041 TI - Prevalence and clinical implications of HFE gene mutations (C282Y and H63D) in patients with chronic hepatitis B and C in Taiwan. AB - BACKGROUND/AIMS: The implication of hemochromatosis (HFE) gene mutations in chronic viral hepatitis remains controversial. We therefore studied the prevalence of HFE mutations and their impact on the progression of chronic viral hepatitis in Taiwan. PATIENTS & METHODS: H63D and C282Y mutations were screened by using polymerase chain reaction followed by restriction fragment length polymorphism in 152 chronic hepatitis B patients with various stages of liver disease, 87 chronic hepatitis C patients with various stages of liver disease, and 49 healthy controls. The distribution of each allele frequency was then compared among different groups of patients and in various stages of liver disease. RESULTS: All three groups of patients were C282Y wild type and the majority of H63D mutations were heterozygotes. Although statistically not significant, allele frequencies of H63D mutation in hepatitis B-related liver cirrhosis (6%) and hepatitis C-related liver cirrhosis (9.1%) were higher than those in healthy control (2%). After adjustment for age and sex, hepatitis B patients with H63D heterozygosity had a higher likelihood of cirrhosis than those with H63D wild type (odds ratios (OR): 3.2, confidence interval (CI): 0.49-20.5, P = 0.22). Similarly, hepatitis C patients with H63D homozygosity had a higher likelihood of cirrhosis compared with those with H63D wild type (OR: 2.35, CI : 0.19-28.5, P = 0.52). CONCLUSIONS: Almost all Taiwanese are C282Y wild type. H63D heterozygote and homozygote, occurring in less than 5% of the subjects, tended to be associated with the development of liver cirrhosis, irrespective of viral etiology. Screening for H63D mutation might be considered in patients with chronic viral hepatitis in Taiwan. PMID- 15780043 TI - SEN virus infection influences the pathological findings in liver but does not affect the incidence of hepatocellular carcinoma in patients with chronic hepatitis C and liver cirrhosis. AB - BACKGROUND/AIMS: This investigation compared the histological findings in the livers of chronic hepatitis C patients who were or were not co-infected with SEN virus (SEN-V) to determine the histological and clinical characteristics of SEN-V infection in Japan. METHODS: Three hundred and ninety-two patients with hepatitis C virus-associated chronic hepatitis (CH) or liver cirrhosis (LC) were included in the study. Serum samples were tested for the presence of SEN-V DNA by nested polymerase chain reaction. The liver biopsy specimen of each patient was examined and scores were assigned to indicate the severity of each of the following features: inflammatory cell infiltration in the periportal, parenchymal, and portal areas; F stage; portal sclerotic change; perivenular fibrosis; pericellular fibrosis; damage to the bile ducts; steatosis and irregular regeneration of hepatocytes (IR). RESULTS: Of the 473 patients, 194 (41.0%) were positive for SEN-V DNA. The rate of progression of F stage correlated with SEN-V DNA positivity. The blood biochemical parameters did not differ significantly between the SEN-V DNA-positive and -negative patients. The histological features of the livers of SEN-V DNA-positive patients included more severe parenchymal inflammatory cell infiltration and more IR. In particular, among those at the F2, F3 and F4 stages, the degree of IR of the SEN-V DNA-positive patients was significantly greater than that of the SEN-V DNA-negative patients. The cumulative probability of hepatocellular carcinoma (HCC) incidence and survival rate did not differ between the SEN-V DNA-positive and-negative patients. CONCLUSIONS: SEN-V co-infection may influence the histopathological features of the livers of patients with type C CH and LC but does not affect the outcome of patients with type C chronic liver disease. PMID- 15780044 TI - Comparison of recurrence after hepatic resection in patients with hepatitis B vs. hepatitis C-related small hepatocellular carcinoma in hepatitis B virus endemic area. AB - PURPOSE: Hepatitis B virus (HBV) and hepatitis C virus (HCV) infection are two important factors in the development of hepatocellular carcinoma (HCC). The carcinogenic mechanism of HBV and HCV is considered to be different. It is interesting to compare the recurrence after hepatic resection in patients with small HCC who were infected with HBV or HCV. METHODS: From 1991 to 1995, 145 patients who were positive for hepatitis B surface antigen (HBsAg) or antibody to HCV (anti-HCV) and diagnosed as small HCC (< or =3 cm) in three medical centers in Taiwan were evaluated in this study. All patients underwent hepatic resection. Among them, 83 (57.2%) were infected by HBV, 51 (35.2%) were infected by HCV, and 11 (7.6%) had dual HBV and HCV infection. RESULTS: Anti-HCV+ HCCs were associated with older age, lower serum albumin, higher alanine transaminase (ALT) level and multi-nodular tumors during diagnosis. During the follow-up, 92 (63.4%) patients developed tumor recurrence. Anti-HCV + HCC had a higher cumulated recurrence rate than HBsAg+ HCC (72.4% vs 53.6 % at 5 year, P = 0.032). In multivariate analysis, the presence of vascular invasion and lower serum albumin levels (<3.9 g/dl) were the determinants for tumor recurrence. CONCLUSIONS: HCV infection, as compared with HBV infection, had a higher cumulated recurrence after hepatic resection in patients with small HCC. Low serum albumin level was significantly associated with recurrence among these patients. PMID- 15780045 TI - Long-term outcome of interferon-alpha-induced autoimmune thyroid disorders in chronic hepatitis C. AB - BACKGROUND: Autoimmune thyroid disorders are among the well-known adverse effects of interferon-alpha (IFN-alpha) therapy in patients with chronic hepatitis C. However, there are few reports regarding the long-term outcome of this complication. We aimed to evaluate the natural history of IFN-alpha-induced autoimmune thyroid disorders with long-term follow-up. METHODS: Four hundred and thirty-nine patients with chronic hepatitis C were treated with IFN-alpha for 24 weeks between March 1993 and April 1998. Seventeen of 439 (3.9%) patients developed symptomatic autoimmune thyroid disorders including nine cases of hyperthyroidism and eight cases of hypothyroidism. The patients with hypothyroidism were all women. These 17 patients were followed up for 71.1 +/- 17.8 months (48-120 months) and were evaluated for long-term outcome. RESULTS: Eight patients could discontinue the thyroid medication (2-36 months, median 10 months). Nine patients needed the thyroid medication at the follow-up period. The patients with hyperthyroidism who needed long-term thyroid medication had a significantly high titer of TSH receptor antibody on onset compared with the patients who could discontinue the thyroid medication. There were no significant differences in age, type of IFN, duration from IFN administration to onset, cessation of IFN, genotype of hepatitis C virus and thyroid hormone levels on onset between the patients who needed long-term thyroid medication and the patients who could discontinue the thyroid medication. CONCLUSION: All patients with IFN-alpha-induced thyroid disorders could be controlled with medication. However, the IFN-alpha-induced thyroid disorders are not always reversible. One must be careful about not only the development of autoimmune thyroid disorders during IFN-alpha therapy but also the outcome of the thyroid disease. PMID- 15780046 TI - Impact of occult hepatitis B virus infection on efficacy and prognosis of interferon-alpha therapy for patients with chronic hepatitis C. AB - BACKGROUND/AIMS: It is reported that some patients with undetectable hepatitis B surface antigen (HBsAg) have serum hepatitis B virus (HBV) DNA in patients with chronic hepatitis C (HCV). The aim of this study was to elucidate the impact of occult HBV infection on the efficacy and prognosis of interferon-alpha (IFN) therapy in HCV patients. METHODS: One hundred and forty HCV patients without HBsAg who received IFN therapy were studied. Serum HBV DNA was quantified by real time detection polymerase chain reaction. RESULTS: Of 140 patients, 11 (7.9%) were HBV DNA-positive before IFN therapy. The serum HBV DNA levels ranged from 106 to 884 copies/ml. Four of these 11 patients showed a sustained virologic response by IFN, compared with 39 of 129 without HBV DNA (P = NS). Interestingly, two of the 11 patients developed hepatocellular carcinoma (HCC) after therapy, compared with 16 of 129 without HBV DNA (P = NS). In the serial study, serum HBV DNA was transiently undetectable during and after IFN; however, most became positive during follow-up. CONCLUSIONS: Occult HBV infection may not have a significant impact on response to IFN therapy for chronic HCV and development of HCC after therapy. Occult HBV may be sensitive to IFN although HBV is not completely eradicated. PMID- 15780047 TI - Immunohistochemical identification of HCV target antigen in paraffin-embedded liver tissue: reproducibility and staining patterns. AB - BACKGROUND: Immunohistochemical staining has been applied successfully to detect hepatitis C virus (HCV) antigen in fresh frozen tissue. In paraffin-embedded tissues, however, minimal trials with conflicting results have been reported. AIMS: The present study is a trial to evaluate the identification of HCV antigen in paraffin-embedded liver biopsies using the anti-HCV monoclonal antibody (MAb) TORDJI-22. METHODS: We applied immunohistochemical staining for HCV in 56 paraffin-embedded liver biopsy specimens, 46 from patients seropositive for HCV RNA and 10 control liver biopsy specimens. The TORDJI-22 MAb was applied in dilution 1:40, with overnight incubation. RESULTS: Reproducible staining patterns of HCV antigen in tissues were identified among the majority (42/46-91%) of HCV RNA seropositive cases. The staining pattern was cytoplasmic of hepatocytes, with occasional nuclear hue. It is mainly coarse granular with microvesicular pattern. Three staining patterns were identified: A, diffuse or membranous; B, patchy; and C, occasional paranuclear. None of the control samples showed a similar staining pattern. CONCLUSION: Immunohistochemical identification of HCV antigen is easy to apply in paraffin-embedded liver biopsy specimens when the optimal detection techniques are applied. The staining pattern is reproducible, being mainly coarse granular cytoplasmic. Cross reactivity with hepatitis B virus antigens was not detected. PMID- 15780048 TI - Radiotherapy for painful bone metastases from hepatocellular carcinoma. AB - BACKGROUND: Painful bone metastases are one of the distressing clinical situations in patients with hepatocellular carcinoma (HCC). The purpose of the study was to evaluate the palliative effect of radiotherapy for painful bone metastases from HCC. PATIENTS AND METHODS: A retrospective analysis was performed upon 51 patients, who were treated with radiotherapy for painful bone metastases from HCC at 77 sites, from January 1991 to June 2000. Twenty patients (39%) presented synchronous metastases, and the remaining 31 patients (61%) showed metachronous metastases. In addition to bone pain, neurologic symptoms and a palpable mass were presented in 13 (25%) and seven (13%) patients, respectively. Twenty-one patients (41%) had a solitary metastasis and the most frequent site was the vertebra. The total dose of radiotherapy ranged from 12.5 to 50 Gy (median 30 Gy). RESULTS: On the Wisconsin Brief Pain Questionnaire, relief of pain was achieved at 56 sites (73%). The overall survival rate at 2 years was 4% and the median survival time was 5 months. Univariate analysis showed that tumor stage within the liver and the presence of metastases to organs, other than bones, were significant prognostic factors for survival (P < 0.05). CONCLUSION: In conclusion, radiotherapy provides effective palliation for patients with painful bone metastases from HCC during the substantial median survival time. PMID- 15780049 TI - High frequency of promoter hypermethylation of RASSF1A in tumor and plasma of patients with hepatocellular carcinoma. AB - PURPOSE: To determine the presence of ras association domain family 1A (RASSF1A) promoter methylation in the tumor tissues and plasma of patients with hepatocellular carcinoma (HCC). EXPERIMENTAL DESIGN: Methylation-specific polymerase chain reaction was used to detect RASSF1A methylation in DNA extracted from HCC tumors and paired plasma samples of 40 patients. The association of RASSF1A hypermethylation in tumor and plasma DNA of HCC patients with clinicopathological characteristics was also analyzed. RESULTS: RASSF1A promoter hypermethylation was detected in 37 of the 40 HCC tissues (92.5%). Of the paired plasma from the 40 HCC patients, aberrant methylation was detected in 17 (42.5%). No RASSF1A methylation was detected in the plasma in the absence of methylation in the corresponding tumor. The presence of RASSF1A promoter hypermethylation in plasma DNA was found to associate with HCC size of > or =4 cm (P = 0.035). CONCLUSION: RASSF1A promoter hypermethylation occurred at a high frequency in HCC. The aberrant methylation was also detectable in over 40% of matched plasma. The latter should be evaluated as a screening tool and/or prognosticator of HCC patients. PMID- 15780050 TI - In vivo and in vitro nuclear magnetic resonance spectroscopy as a tool for investigating hepatobiliary disease: a review of H and P MRS applications. AB - Nuclear magnetic resonance (NMR) spectroscopy is a non-invasive technique, which allows the study of cellular biochemistry and metabolism. It is a diverse research tool, widely used by biochemists to investigate pathophysiological processes in vitro and, more recently, by physicians to determine disease abnormalities in vivo. This article reviews the basics of the NMR phenomenon and summarises previous research on the hepatobiliary system using both laboratory based and clinical methodologies. The role of proton and phosphorus-31 ((31)P) NMR spectroscopy in the study of malignant and non-malignant liver disease and studies of bile composition are discussed. In vivo techniques (magnetic resonance spectroscopy, MRS) can be performed as an adjunct to standard MR examination of the liver. Although still primarily a research tool, the in vivo technique provides non-invasive biochemical information on disease severity and holds promise in its use to gauge response to treatment regimens. PMID- 15780051 TI - Physical hemodynamic interaction between portal venous and hepatic arterial blood flow in humans. AB - OBJECTIVE: The hepatic arterial end-diastolic velocity (HAEDV) is normally equal to portal vein peak velocity (PVPV). However, there is no report of quantitative measurement that HAEDV was equal to PVPV. We investigated the interaction in PVPV and HAEDV in both chronic and acute hepatic hemodynamic changes. METHODS: One hundred and nineteen patients (54 with cirrhosis, 23 with chronic hepatitis, and 42 with no diffuse liver disorder) were enrolled. We investigated the differences in PVPV and HAEDV among the patients with and without liver disorder. In addition, we measured the intraindividual changes in HAEDV when PVPV was mechanically changed by percutaneous isolated hepatic perfusion in six patients and by percutaneous transhepatic portal embolization (PTPE) in six more. RESULTS: HAEDV was nearly equal to PVPV not only in patients with both normal and hepatitis but also in those with cirrhosis (PVPV-HAEDV = 3.0 +/- 5.2, 2.2 +/- 5.4, 2.3 +/- 6.5 cm/s, respectively). In the intraindividual study, both PVPV and HAEDV decreased during hepatic mechanical perfusion and HAEDV was equal to PVPV (8.2 +/- 2.8, 10.5 +/- 1.5 cm/s, respectively). After PTPE, PVPV was increased and hepatic arterial peak systolic velocity was reciprocally decreased. However, HAEDV was nearly equal to PVPV 7 days after PTPE (PVPV-HAEDV = 5.9 +/- 5.1 cm/s). CONCLUSIONS: Since arterial end-diastolic velocity depends on the downstream vascular resistance, lower HAEDV in patients with cirrhosis was considered to reflect a high outflow resistance. If there is no collateral pathway, we consider that HAEDV may actually reflect sinusoidal resistance to the same degree as PVPV. PMID- 15780052 TI - Signal intensity of the liver parenchyma in microbubble contrast agent in the late liver phase reflects advanced fibrosis of the liver. AB - BACKGROUND: Microbubble of Levovist accumulates in liver parenchyma, and the phenomenon has been reported as late liver-specific parenchymal. The aim of the present study was to compare the parenchymal enhancement effect of Levovist with the degree of liver dysfunction. PATIENTS AND METHODS: Sixty consecutive patients who consented to be treated were enrolled in this study. Pulse-inversion ultrasonography (US) in the liver parenchymal phase of enhancement with Levovist was performed in a preoperative examination. The mechanical index of pulse inversion US was set at 1.3. The gray-scale intensity of the non-tumor area of the liver parenchyma at the level of the focal zone was measured. The hepatic fibrosis index was measured in each liver by morphometric analysis. The correlation between the gray-scale intensity of the non-tumor area of the liver parenchyma and the hepatic fibrosis index was assessed. RESULTS: There was a significant inverse correlation between the gray scale of the liver parenchyma and the hepatic fibrosis index (r = -0.809, P < 0.01). The average signal intensity of the liver parenchyma was 144.5 in a normal liver, 133.6 in chronic hepatitis, and 102.6 in liver cirrhosis, demonstrating a significant difference between a normal and cirrhotic liver (P < 0.01). CONCLUSIONS: The signal intensity of a microbubble disruption of the liver parenchyma in the late phase of enhancement with Levovist was considered to reflect the degree of hepatic fibrosis. PMID- 15780053 TI - Proposal for a histological scoring and grading system for non-alcoholic fatty liver disease. AB - AIM/BACKGROUND: The spectrum of histopathological features in non-alcoholic fatty liver disease (NAFLD) has been well described. At least two scoring systems have been established. We propose here a system in which numerical scores are obtained using the different features. METHODS: Twenty-five cases of well-defined NAFLD were identified. Two pathologists blinded to identifiers and clinical data independently scored the liver biopsies twice for portal fibrosis (PF: 0-6), lobular inflammation and necrosis (LIN: 0-3), Mallory bodies (MB: 0-3), hepatocyte ballooning (HB: 0-3), perisinusoidal fibrosis (PSF: 0-3) and fatty change (FC: 1-4). The kappa statistic tested observer concordance. Non-parametric measures of correlation and hierarchical cluster analysis were used to elaborate a grading system. RESULTS: A broad spectrum of NAFLD was observed. Intra- and interobserver concordance was satisfactory. An activity score was created (AS: 0 12) as the sum of LIN, MB, HB and PSF, but not FC. A system for severity of NAFLD was developed: Grade 1 (PF: 0-2 and AS: 0-4), Grade 2 (PF: 3 or AS: 5-7) and Grade 3 (PF: 4-6 or AS: 8-12). Diabetes, elevated alkaline phosphatase and decreased platelets were associated with advanced grade. CONCLUSIONS: This simple, reproducible NAFLD score produces a three-tier severity grade. This numerical system may prove useful in assessing disease severity and interval changes. PMID- 15780054 TI - Osteoprotegerin and RANKL in alcoholic liver cirrhosis. AB - BACKGROUND/AIMS: The mechanisms leading to osteoporosis in alcoholic liver disease remain poorly understood. Recently identified soluble circulating osteoprotegerin (OPG), is the osteoclastogenesis inhibitory factor. It acts as a decoy receptor for osteoclast activating factor, receptor activator of nuclear factor-kappaB ligand (RANKL), and impairs osteoclast function. The aim of our study was to investigate the OPG/RANKL system in alcoholic cirrhotic patients and their correlation with biochemical marker of bone turnover. PATIENTS AND METHODS: Serum OPG, RANKL, osteocalcin (OC), C-terminal cross-linking telopeptide of type I collagen (CTX-I), bone alkaline phosphatase activity (bALP), and urinary hydroxyproline were measured in 30 patients with alcoholic cirrhosis, and in 20 age- and sex-matched healthy controls. RESULTS: OPG levels were significantly increased in patients with alcoholic cirrhosis compared with healthy subjects (5.9 pmol/l, range 2.7-9.0 vs 4.1 pmol/l, range 1.2-6.6; P < 0.001). RANKL levels were significantly higher in patients with cirrhosis (0.48 pmol/l, range 0.01 1.34) than in healthy subjects (0.11 pmol/l, range 0.01-0.90). There was a positive correlation between serum OPG and RANKL (r = 0.37; P < 0.001), bALP (r = 0.66; P < 0.001) and urinary hydroxyproline (r = 0.51; P < 0.05) but not with OC and CTX-I. CONCLUSIONS: OPG might partly represent a compensating mechanism to the negative balance of bone remodelling in patients with alcoholic cirrhosis. PMID- 15780055 TI - Severity of liver disease does not predict osteopenia or low bone mineral density in primary sclerosing cholangitis. AB - BACKGROUND: The association between metabolic bone disease and cholestatic liver disease has been poorly characterized. To date a single institution has published data suggesting that in primary sclerosing cholangitis (PSC), advanced liver disease predicts advanced bone disease. AIM: To determine the association between the severity of liver disease and bone mineral density (BMD) in PSC patients. METHODS: We identified 30 PSC patients who had undergone dual energy X-ray absorptiometry (DXA) scan. We compared lumbar spine DXA scores between patients with more and less advanced liver disease. RESULTS: Nine patients were osteopenic (30%), and one patient was osteoporotic. Five patients were female (17%), and none was postmenopausal. BMD was not different between patients listed and not listed for liver transplantation (P = 0.49) or between patients with and without hepatic decompensation (P = 0.63). Model for end-stage liver disease (MELD) score (P = 0.99) and the modified Mayo risk score (P = 0.25) did not predict BMD. CONCLUSIONS: Our study is the first to suggest that low bone density cannot be predicted by the severity of liver disease in PSC. Perhaps other known risk factors for osteoporosis will be important predictors of abnormal bone density in this patient population. PMID- 15780056 TI - Does the diagnosis of primary biliary cirrhosis or autoimmune cholangitis depend on the 'phase' of the disease? AB - BACKGROUND: It is unclear whether autoimmune cholangitis (AIC) is a separate disease entity or primary biliary cirrhosis (PBC) without antimitochondrial antibodies (AMA) since fluctuation of AMA titres by immunofluorescence (IF) is often observed during the course of PBC. The aim of this study was to determine the serial changes in AMA profiles during the course of initially diagnosed PBC or AIC. METHODS: In this prospective study, 32 patients with PBC or AIC were followed-up for at least 20 months and tested for AMA by IF, enzyme-linked immunosorbent assay (ELISA), and immunoblotting (IB). RESULTS: When positive AMA result was defined as 'AMA by IF positive', 'AMA by IF and/or ELISA positive', and 'AMA by IB positive', the diagnosis of PBC or AIC did not change in 78%, 91%, and 97%, respectively, throughout follow-up. However, the diagnosis changed in one patient, and three patients were diagnosed as AIC throughout follow-up, despite the use of all three assays. CONCLUSIONS: Our results suggested that the diagnosis of PBC and AIC was dependent on the 'phase' of the respective disease in 22% of the patients when negative AMA result was defined as 'AMA by IF negative'. This may result in recommending IB analysis before making the diagnosis of AIC. PMID- 15780057 TI - An unusual form of autoimmune hepatitis in young Somalian men. AB - BACKGROUND: Significant diversity in disease severity has been identified for autoimmune disorders among different ethnic groups. Current knowledge of both the natural history and management of autoimmune hepatitis (AIH) has been derived from European or Japanese patients, and there is limited information about the disease in patients from other ethnic groups. AIMS: To assess the clinical, histological and immunological features of AIH in patients from Somalia and to determine their response to therapy. METHODS: Retrospective review of a cohort of young Somalian men with atypical AIH compared with a control group of European patients. RESULTS: The six Somalian men were younger at presentation (median age 37 (range 24-59) years) than the seven female and three male European controls (55 (34-54) years, P = 0.06). The Somalians had slightly more severe disease at presentation-median modified Ishak stage of 2.5 compared with 2 in Europeans (P = 0.61) and four (66%) had features of cholestasis compared with only one (10%) European patient (P = 0.04). Therapy with prednisolone and azathioprine was completely effective for eight of 10 Europeans but only one of seven Somalians (P = 0.04). Analysis of human leucocyte antigen types revealed differences between the Somalian and European patients, although these differences did not reach statistical significance. CONCLUSIONS: Somalian men with AIH present with cholestatic features and respond poorly to standard immunosuppressive regimes. PMID- 15780058 TI - Reduced prevalence of ischemic events and abnormal supraortic flow patterns in patients with liver cirrhosis. AB - BACKGROUND: A reduced prevalence of cardiovascular diseases has been reported in liver cirrhosis. However, studies focusing on supraortic district of cirrhotic patients are lacking. METHODS: By ultrasound duplex scanning, the presence and severity of atherosclerotic plaques and flow pattern abnormalities were assessed in carotid and vertebral arteries of 118 cirrhotics aged 60.7 +/- 12.8 (1 standard deviation) years, and in 236 controls matched with cirrhotic patients according to age, sex and cigarette smoking. RESULTS: Previous ischemic strokes were significantly less numerous in cirrhotic patients than in controls (0.8% vs. 10.5%; P = 0.0009); also the prevalence of myocardial infarction was significantly reduced (1.7% vs. 6.4%; P = 0.0532). Moreover, cirrhotic patients differed from controls for a lower prevalence of hypertension and hypercholesterolemia, and for a greater proportion of diabetics. Although the presence and severity of atherosclerotic plaques was similar in the two groups, liver cirrhosis was associated with a lower prevalence of abnormal flow patterns (13.6% vs. 29.2%; P = 0.0011). The inverse association of hemodynamic changes with liver cirrhosis persisted after all main risk factors were simultaneously taken into account by multiple logistic regression. However, in the presence of hypertension, hypercholesterolemia and cigarette smoking, the 'protective' effect of cirrhosis on the occurrence of abnormal flow patterns was no longer detectable. CONCLUSIONS: Advanced liver disease is associated with a reduced prevalence of ischemic stroke, which seems to be related to a decreased prevalence of abnormal flow patterns in the supraortic vessels, especially among non-smokers. PMID- 15780059 TI - Intrahepatic circulatory time analysis of an ultrasound contrast agent in liver cirrhosis. AB - OBJECTIVE: To assess the diagnostic accuracy and advantages of intrahepatic circulatory time analysis of an ultrasound contrast agent using pulse-inversion imaging as compared with recently reported noninvasive diagnostic tests for cirrhosis. METHODS: Forty patients divided into noncirrhotic (nonLC) (n = 20) and compensated cirrhotic (LC) (n = 20) groups were studied prospectively. After intravenous administration of a contrast agent, the arrival times at the hepatic artery (HA), portal vein (PV), and hepatic vein (HV) were measured by pulse inversion imaging. Intrahepatic circulatory time was calculated as the difference between the HV and HA arrival times (HV-HA interval time) or the HV and PV arrival times (HV-PV interval time). RESULTS: The HV-HA and HV-PV interval times were significantly shorter in the LC group (7.4 +/- 1.7 and 1.9 +/- 1.5 s, respectively) compared with those in the nonLC group (normal: 15.6 +/- 2.1 and 11.1 +/- 1.7 s, respectively; P < 0.001 and P < 0.001, respectively, and hepatitis: 12.8 +/- 4.1 and 7.8 +/- 4.4 s, respectively; P < 0.001 and P < 0.002, respectively). Each intrahepatic circulatory time showed the highest accuracy rate for cirrhosis in other noninvasive diagnostic tests. CONCLUSIONS: This analysis, which is considered to reflect intrahepatic hemodynamic changes, is a useful noninvasive diagnostic test for compensated cirrhosis. PMID- 15780060 TI - Instant diagnosis of spontaneous bacterial peritonitis using leukocyte esterase reagent strips: Nephur-Test vs. MultistixSG. AB - BACKGROUND/AIM: Spontaneous bacterial peritonitis (SBP), defined by an ascites neutrophil count >250/mul, is a severe complication of cirrhosis and demands prompt diagnosis. Leukocyte esterase reagent strips can rapidly detect leukocytes in physiological fluids. We compared the performance of two leukocyte esterase reagent strips in the diagnosis of SBP. METHODS: Non-selected patients with cirrhosis and ascites were prospectively included in a Community Hospital. Manual ascites cytology, culture and BIOCHEMISTRY were compared with the Nephur-Test (read at 60 s: negative + 1 to + 3) and MultistixSG (read at 120 s: negative, trace, +1 to +3). Multiple investigators performed the tests with repeat readings by one nurse. RESULTS: Two hundred and forty five ascites fluid samples were obtained in 51 patients: 44% in conditions of SBP antibiotic prophylaxis, 64% in an ambulatory setting. Seventeen samples had an ascites polymorphonuclear count of > or =250/microl, associated with a positive ascites culture in seven, obtained in 12 patients (24%), 53% in an ambulatory setting. Sensitivity, specificity, and positive and negative predictive values were: Nephur-Test (88.2%, 99.6%, 93.8%, 99.1%); MultistixSG (64.7%, 99.6%, 91.7%, 97.4%). There was excellent agreement between the investigators and the nurse. CONCLUSIONS: Leukocyte esterase reagent strips may provide a rapid bedside diagnosis of SBP and thereby shorten time to appropriate management. The Nephur-Test appeared to out-perform MultistixSG. These tests might be particularly useful in ambulatory settings or whenever ascites fluid analysis is not rapidly available. PMID- 15780061 TI - Transjugular intrahepatic portosystemic shunt in refractory ascites: a meta analysis. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is a more effective treatment for refractory ascites than large volume paracentesis (LVP), but the magnitude of its effect in terms of control of ascites, encephalopathy and survival has not been established. AIM: This meta-analysis compare TIPS to LVP in terms of control of ascites at 4 and 12 months, encephalopathy and survival at 1 and 2 years. RESULTS: Five randomized controlled trials involving 330 patients were included. In the TIPS group, control of ascites was more frequently achieved at 4 months (66% vs 23.8%, mean difference: 41.4%, 95% confidence interval (CI): 29.5-53.2%, P < 0.001) and 12 months (54.8% vs 18.9%, mean difference: 35%, 95% CI: 24.9-45.1%, P < 0.001), whereas encephalopathy was higher (54.9% vs 38.1%, mean difference: 17%, 95% CI: 7.3-26.6%, P < 0.001). Survival at 1 year (61.7% vs 56.5%, mean difference: 3.2%, 95% CI: -14.7 to 21.9%) and 2 years (50% vs 42.8%, mean difference: 6.8%, 95% CI: -10 to 23.6%) were not significantly different. CONCLUSIONS: TIPS is a more effective treatment for refractory ascites than LVP. However, TIPS increase encephalopathy and does not improve survival. PMID- 15780062 TI - In situ expression patterns of melanoma inhibitory activity 2 in healthy and diseased livers. AB - Recently we identified a novel gene of the MIA gene family, melanoma inhibitory activity 2 (MIA2) and found that MIA2 mRNA is selectively expressed in hepatocytes. Here, we analyzed the in situ expression of MIA2 protein and mRNA in healthy and diseased livers to get first insights into the function of MIA2. METHODS: We analyzed liver tissue of patients with chronic hepatitis C (HepC) infection and hepatocellular carcinoma (HCC) as well as a human multi-tissue array, primary human hepatocytes and the hepatoma cell-lines HepG2, Hep3B and PLC by immunohistochemical staining and quantitative RT-PCR. In addition to MIA2, the expression of alpha-smooth muscle actin (alpha-sma), a marker for activated hepatic stellate cells (HSCs)/myofibroblast, was analyzed. RESULTS: Hepatocytes were confirmed as the exclusive cellular source of MIA2 expression, with a granular, cytoplasmatic staining pattern without enhancement at the cell membrane. In contrast, only low MIA2 expression levels were detected in most HCC and hepatoma cell lines. Only in HCC that contained fibrous stroma or thick hyalinized bundles, adjacent atypical hepatocytes revealed strong staining. In accordance, MIA2 expression was also upregulated in non-tumorous livers of patients with HepC and correlated with the staging of fibrosis. Interestingly, both in HCC and liver tissues of patients with HepC we found a correlation of MIA2 and alpha-sma expression. DISCUSSION: We define for the first time in situ expression patterns of MIA2 in healthy and diseased livers. Our data raise the hypothesis that activation of HSCs/myofibroblasts has influence on MIA2 expression in vivo, consistent with our previous in vitro findings. Since the staining pattern and the protein structure highly suggests that MIA2 is a secreted protein, it may possibly serve as a marker of hepatic fibrosis. PMID- 15780063 TI - Hepatobiliary transporter expression in human hepatocellular carcinoma. AB - BACKGROUND/AIMS: Treatment of hepatocellular carcinoma (HCC) is hampered by resistance to chemotherapy, which might be mediated by multidrug resistance P glycoproteins (MDR P-gps) and MDR-associated proteins (MRPs). The effectiveness of cytostatics could be further impeded by reduced hepatocellular drug uptake into HCCs. Therefore, we aimed to determine P-gp, MRP and organic anion transporting protein OATP2 (SLC21A6) expression in HCC. Furthermore, we investigated expression of the major bile salt uptake system Na(+)/taurocholate cotransporter NTCP (SLC10A1), since bile salt-coupled chemotherapeutics were proposed to increase therapeutic drug enrichment in HCC. MATERIAL/METHODS: mRNA and protein expression and tissue distribution of P-gps, MRPs, OATP2 and NTCP were assessed in HCC and peritumorous non-neoplastic tissue by reverse transcription polymerase chain reaction, Western blotting and immunohistochemistry, respectively. RESULTS: Expression of P-gps (multidrug export pump MDR1 (ABCB1), phospholipid flippase MDR3 (ABCB4), sister of P glycoprotein SPGP (ABCB11)) and basolateral MRP homologue MRP3 (ABCC3) showed a trend for decreased levels in HCC but was highly variable among individual tumors. In contrast, canalicular conjugate export pump MRP2 (ABCC2) expression was generally maintained or even showed a trend towards increased levels. NTCP and OATP2 expression was markedly reduced in most HCCs (P < 0.05). Expression of the genuine drug transporter, the concentrative nucleoside transporter (CNT1), was highly variable and showed a trend for reduced levels in HCC. SUMMARY/CONCLUSIONS: MRP2 seems to be the major candidate transporter involved in chemoresistance and reduced expression of OATP2 may further contribute to low drug accumulation in HCCs. Overexpression of drug exporters is not a general feature of HCC but could account for chemoresistance of individual cases. Since expression of uptake systems is generally reduced in HCC, bile salt-coupled therapeutics may not represent a suitable strategy to overcome insufficient drug enrichment. PMID- 15780064 TI - Decreased expression and frequent allelic inactivation of the RUNX3 gene at 1p36 in human hepatocellular carcinoma. AB - BACKGROUND/AIMS: Alteration in transforming growth factor-beta signaling pathway is one of the main causes of hepatocellular carcinoma (HCC). The human runt related transcription factor 3 gene (RUNX3) is an important component of this pathway. RUNX3 locus 1p36 is commonly deleted in a variety of human cancers, including HCC. Therefore, we examined genetic and epigenetic alterations of RUNX3 in human HCC. METHODS: Five HCC cell lines and 41 patients with HCC were investigated in this study. We examined the expression of RUNX3 mRNA, methylation status of RUNX3 promoter region, loss of heterozygosity (LOH) at 1p36, and mutation analysis. These results were compared with clinicopathological data. RESULTS: Promoter hypermethylation was detected in four (80%) of five HCC cell lines and 31 (75.6%) of 41 HCC tissues, confirmed by sequence of bisulfite treated DNA. LOH was detected in 14 (37.8%) of 37 HCC. By comparison with clinicopathological data, hypermethylation was more common in hepatitis C virus antibody and formation of capsule-positive cases, and decrease of expression was correlated strongly with advanced stage and LOH-detected cases. CONCLUSION: Hypermethylation and LOH appear to be common mechanisms for inactivation of RUNX3 in HCC. Therefore, RUNX3 may be an important tumor suppressor gene related to hepatocarcinogenesis. PMID- 15780065 TI - Liver cell lines for the study of hepatocyte functions and immunological response. AB - BACKGROUND: Liver cell lines closely resembling primary hepatocyte are essential for research on hepatitis viruses and hepatocyte function. Currently used cell lines are derived from hepatic tumours and have altered gene expression. AIMS: The generation and characterisation of novel human hepatocyte lines (HHLs) derived from healthy human liver, retaining the primary hepatocyte phenotype. RESULTS: Primary hepatocytes were immortalised with Moloney's mouse leukaemia virus expressing E6 and E7 proteins of human papillomavirus, and cultures propagated long-term. All HHLs contained markers of hepatocyte and biliary phenotype (cytokeratins 7, 8, 18 and 19), Cytochrome P450 and albumin. The HHLs did not express high levels of p53 or alpha-fetoprotein. When grown in a collagen sandwich culture, or at the air-liquid interface, HHLs were maintained as monolayer whereas Huh-7 and HepG2 formed thick layers. All HHLs showed increased capacity to bind recombinant hepatitis C virus-like particles in comparison with Huh-7 and HepG2. We also demonstrate that HHLs contained active gap junctions, and that the cells respond to stimulation with IFN-alpha by upregulation of major histocompatibility complex (MHC)-I and -II. CONCLUSIONS: These HHLs retain primary hepatocyte phenotype and should be useful for investigating mechanisms of entry and replication of hepatotropic viruses, and should also be valuable in the study of hepatocyte biology and pathology. PMID- 15780066 TI - Cyclic AMP-phosphodiesterases inhibitor improves sodium excretion in rats with cirrhosis and ascites. AB - BACKGROUND: The mechanisms responsible for renal dysfunction and sodium retention in cirrhosis remain unclear. Cyclic AMP (cAMP) regulates sodium reabsorption in the proximal nephron. This study investigates the role of cAMP metabolism in renal dysfunction in cirrhosis. METHODS: Renal function was studied by the clearance technique in anesthetized control and cirrhotic rats with or without ascites. cAMP phosphodiesterase (PDE) activity was measured in the renal cortex in vitro. Moroever, the effects on renal function of the intravenous administration of cAMP and rolipram, a powerful and specific cAMP-PDE4 inhibitor, were evaluated. RESULTS: In control and in non-ascitic cirrhotic rats, cAMP administration significantly increased sodium and phosphate excretions, but did not change these excretions in cirrhotic rats with ascites. cAMP-PDE activity was higher in ascitic than in control rats (P < 0.05). Rolipram infusion significantly increased sodium and phosphate excretion only in cirrhotic rats with ascites. CONCLUSION: These results suggest that increased renal cAMP-PDE activity is responsible for resistance to the natriuretic effects of cAMP in cirrhosis and plays a role in the development of ascites. PMID- 15780067 TI - Anti-tRNP(ser)sec/SLA/LP autoantibodies. Comparative study using in-house ELISA with a recombinant 48.8 kDa protein, immunoblot, and analysis of immunoprecipitated RNAs. AB - BACKGROUND: Antibodies against tRNP((ser)sec) (ribonucleoproteins, RNP) have been described in our laboratory as markers of poor outcome in type 1 autoimmune hepatitis (AIH). The antigenic protein has been sequenced and cloned as a 48.8 kDa protein and identified with soluble liver antigen (SLA) and liver-pancreas (LP) antigen. The aim of this paper was to determine the best assay by which to detect these antibodies in type 1 AIH. METHODS: A simple and reliable enzyme linked immunoassay based on prokaryotically expressed protein was compared with an immunoblot assay using prokaryotically- and eukaryotically-expressed proteins and an assay based on immunoprecipitated RNAs from HeLa cell extracts. Eighty-one sera from 58 patients with type 1 AIH, 168 sera from patients with autoimmune diseases or chronic hepatitis C, and 60 sera from healthy subjects were similarly tested. RESULTS: The specificity of the assays was 100%, but the frequency of seropositivity was higher in the assay based on immunoprecipitated RNAs (44.4%) than in the enzyme-linked immunosorbent assay (ELISA) (16%) and the immunoblot assay with prokaryotically (12.34%) and eukaryotically (14.8%)-expressed protein. There were no clinical differences between the patients positive by ELISA, immunoblot assay, or immunoprecipitated RNAs. CONCLUSIONS: These results suggest that the analysis of the immunoprecipitated RNAs is the most useful, sensitive and specific method to detect anti-tRNP((ser)sec)/SLA/LP autoantibodies. PMID- 15780068 TI - Effect of IL-4 and IL-13 on collagen production in cultured LI90 human hepatic stellate cells. AB - BACKGROUND: Recently, it has been reported that interleukin 4 (IL-4) and 13 (IL 13) directly activate fibroblasts and promote fibrosis. In the process of hepatic fibrosis, the effects of these cytokines on hepatic stellate cells (HSCs) are not well known. METHODS: We evaluated the effects of IL-4 and IL-13 on the collagen production and the proliferation of LI90, a hepatic stellate cell line. We also examined whether interferon (IFN) interferes with the expression of collagen, since IFN has been reported to clinically suppress hepatic fibrosis. RESULTS: The receptor complex for IL-4 and IL-13 was IL-4Ralpha/IL-13Ralpha1 on LI90 cells, and the phosphorylation of Stat6 was induced by IL-4 and IL-13. The treatment of LI90 cells with IL-4 or IL-13 increased the production of collagen I protein levels by nearly three times in comparison with untreated cells. Collagen mRNA levels were increased roughly 10-fold by IL-4 and 100-fold by IL-13. Interestingly, BrdU incorporation in LI90 cells was decreased by IL-4 or IL-13 treatment. Furthermore, induction of collagen I production by these cytokines was blocked by IFNalpha or IFNbeta treatment, although neither treatment alone suppressed collagen production. CONCLUSIONS: Our data suggested that IL-4 and IL 13 directly affected HSCs by increasing collagen production and suppressing cell proliferation. The anti-fibrogenetic effect of IFN may be due in part to the blockade of IL-4 and IL-13 stimulation of HSCs. PMID- 15780069 TI - The role of nitric oxide in the pathogenesis of systemic and splanchnic vasodilation in cirrhotic rats before and after the onset of ascites. AB - BACKGROUND: The role of nitric oxide (NO) in the pathogenesis of splanchnic arterial vasodilation in cirrhosis has been recently debated by some experimental studies. AIMS: We investigated the role of NO in the pathogenesis of the splanchnic arterial vasodilation along the course of CCl(4)-induced experimental cirrhosis. METHODS: We analyzed the effect on mean arterial pressure (MAP), cardiac output (CO), total peripheral resistance (TPR), and resistance in the superior mesenteric artery (RSMA), before and after the administration of a unspecific NO synthase (NOS) inhibitor (Nomega-nitro-L-arginine-methyl-ester, L NAME) and a specific NOS2 inhibitor (L-N-(1-iminoethyl)-lysine, L-NIL) to cirrhotic rats with and without ascites, and to control rats. NOS2 and NOS3 protein expression was also assessed in systemic and splanchnic arteries of these animals. RESULTS: L-NAME in cirrhotic rats markedly improved MAP, and TPR and decreased CO regardless of whether they had ascites or not. L-NIL did not produce any significant effect on systemic haemodynamics in control and cirrhotic rats. NOS3 overexpression in the aorta of cirrhotic animals paralleled the progression of the liver disease. L-NAME increased RSMA in cirrhotic rats, but this effect was much less intense in rats with ascites. L-NIL had an effect only on RSMA in rats with ascites, which was of a similar extent to that produced by L-NAME. Western blot experiment showed a faint overexpression of NOS3 in the mesenteric artery of cirrhotic rats with and without ascites and a clear induction of NOS2 only in the mesenteric artery of rats with ascites. CONCLUSIONS: These results indicate that NO contributes significantly to the pathogenesis of arterial splanchnic circulation in the early stages of experimental cirrhosis but has only a minor role in its maintenance after the development of ascites. Furthermore, the expression of the different NOS isoforms varies along the course of the liver disease. PMID- 15780070 TI - Effects of prior splenectomy on remnant liver after partial hepatectomy with Pringle maneuver in rats. AB - BACKGROUND/AIMS: In the case of the liver resection, the temporary occlusion of the hepatoduodenal ligament (Pringle maneuver) is often used. However, the maneuver causes hepatic ischemia/reperfusion (I/R) injury that strongly affects the recovery of patients. The present study investigated the effects of prior splenectomy on the remnant liver in partial hepatectomized rat with Pringle maneuver. METHODS: Pringle maneuver was conducted just before a two-thirds partial hepatectomy. Efficacy of splenectomy was assessed by survival rate, serum alanine aminotransferase (ALT), neutrophil infiltration into liver, recovery of remnant liver weight, and liver proliferating cell nuclear antigen (PCNA) levels. Ischemic preconditioning was performed as follows; 10 min of total hepatic ischemia followed by 10 min of reperfusion. RESULTS: In partial hepatectomized rats with 30 min of Pringle maneuver, seven out of 12 rats died within 3 days. On the other hand, when splenectomy was performed on 3 days before the maneuver, only one out of 12 rats died. When prior splenectomy was performed on eight and 18 days before the Pringle maneuver, respectively, similar efficacy was observed. In addition, prior splenectomy on 3 days before the maneuver showed that serum ALT activity, neutrophil infiltration, recovery of remnant liver weight, and PCNA levels in partial hepatectomized rats with Pringle maneuver were also ameliorated as compared with those of control rats without splenectomy. When effects of prior splenectomy were compared with those of ischemic preconditioning in these situations, efficacy of prior splenectomy was comparable with that of the ischemic preconditioning. CONCLUSIONS: Prior splenectomy ameliorated the I/R injury in the remnant liver after partial hepatectomy with Pringle maneuver. Effects of prior splenectomy may influence the liver for long duration, because splenectomy on 18 days before the maneuver still exerts effective action. PMID- 15780071 TI - Interleukin-6 inhibits cell proliferation in a rat model of hepatocellular carcinoma. AB - BACKGROUND: Interleukin-6 (IL-6) plays a critical role in normal hepatic growth and liver regeneration. The aims of the present study are to determine the expression of components of IL-6 signaling in an in vivo model of hepatocellular carcinoma (HCC) and address the role of IL-6 signaling in the progression of HCC. METHODS: An in vivo rat HCC model was established and IL-6 receptor (IL-6R) and downstream signaling pathway expression and activity were determined in HCC and normal liver specimens. Tumorigenic HCC cells from resected HCC samples and normal hepatocytes were then isolated and cultured in the presence and absence of recombinant human IL-6 (rhIL-6). RESULTS: HCC specimens demonstrated decreased IL 6Ralpha/gp130 expression as compared with the normal liver. In contrast, HCC samples had significantly increased IL-6 messenger RNA expression and signal transducers and activators of transcription (STAT)3 activity. Using in vitro cell cultures, we demonstrated that IL-6 stimulated STAT3 and extracellular regulated kinase (ERK) activity in both HCC cells and isolated hepatocytes. However, while STAT3 activation profiles were similar, IL-6 stimulated ERK activity in a biphasic manner in HCC cells and a monophasic, sustained ERK activation in hepatocytes. In HCC cells, a significant induction of cyclin-dependent kinase (CDK) inhibitors, p21(waf1/cip1) and p27(Kip1) occurred, an effect that was not observed in normal hepatocytes. Finally, we established that IL-6 acted to inhibit serum-stimulated DNA synthesis and cell mitogenesis in HCC cells in vitro. CONCLUSIONS: These data demonstrate altered expression of components of IL 6 signaling in HCC in vivo. IL-6 treatment of HCC cells inhibits serum-stimulated mitogenesis, possibly via differences in activation profiles of intracellular signaling pathways and their effect on CDK inhibitor expression/activity. PMID- 15780072 TI - Primary hepatic nerve sheath tumor. PMID- 15780075 TI - IL-10, IL-6, and TNF-alpha: central factors in the altered cytokine network of uremia--the good, the bad, and the ugly. AB - It has been increasingly apparent that wasting and cardiovascular disease (CVD) is associated with a persistent systemic inflammatory response in end-stage renal disease (ESRD) patients. The reasons for the increased risk of inflammation in ESRD patients appear to be complex, including non-dialysis as well as dialysis related factors. The combination of an impaired immune response coupled with persistent immune stimulation may have a role in the low-grade systemic inflammation and altered cytokine balance that characterizes the uremic state and which may translate into increased risk for vascular disease. The accelerated atherosclerotic process of ESRD may involve several interrelated processes, such as oxidative stress, endothelial dysfunction, and vascular calcification, in a milieu of constant low-grade inflammation with impaired function of neutrophils and T cells, as well as a dysregulated cytokine network. Although a large number of pro- and anti-inflammatory cytokines are of importance, available data suggest that the anti-inflammatory cytokine interleukin (IL)-10 and the mainly proinflammatory cytokines IL-6 and tumor necrosis factor-alpha (TNF-alpha) may play important roles in the development of Th imbalance, CVD and wasting in the uremic milieu. Given the strong association between proinflammatory cytokines and complications common in ESRD, such as vascular calcification and wasting, the potential role of both general and targeted anticytokine treatment strategies in ESRD patients needs further evaluation. PMID- 15780076 TI - Molecular pathogenesis of ADPKD: the polycystin complex gets complex. AB - Autosomal-dominant polycystic kidney disease (ADPKD) is one of the most common human monogenic diseases with an incidence of 1:400 to 1:1000. It is characterized by the progressive development and enlargement of focal cysts in both kidneys, typically resulting in end-stage renal disease (ESRD) by the fifth decade. The cystogenic process is highly complex with a cellular phenotype consistent with "dedifferentiation" (i.e., a high proliferative rate, increased apoptosis, altered protein sorting, changed secretory characteristics, and disorganization of the extracellular matrix). Although cystic renal disease is the major cause of morbidity, the occurrence of nonrenal cysts, most notably in the liver (occasionally resulting in clinically significant polycystic liver disease) and the increased prevalence of other abnormalities including intracranial aneurysms, indicate that ADPKD is a systemic disorder. Following the identification of the first ADPKD gene, PKD1, 10 years ago and PKD2 2 years later, considerable progress has been made in defining the etiology and understanding the pathogenesis of this disorder, knowledge that is now leading to the development of several promising new therapies. The purpose of this review is to summarize our current state of knowledge as to the structure and function of the PKD1 and PKD2 proteins, polycystin-1 and -2, respectively, and explore how mutation at these loci results in the spectrum of changes seen in ADPKD. PMID- 15780077 TI - Analysis of NPHS1, NPHS2, ACTN4, and WT1 in Japanese patients with congenital nephrotic syndrome. AB - BACKGROUND: Congenital nephrotic syndrome (CNS) causes significant renal failure, and is classified into two types: (1) Finnish type; and (2) other, including diffuse mesangial sclerosis. Mutations of NPHS1 and NPHS2, which encode the slit diaphragm components nephrin and podocin, cause CNS and autosomal-recessive familial steroid-resistant nephrotic syndrome, respectively. Most patients with Finnish-type CNS in Europe and the United States have NPHS1 mutations. However, NPHS2 mutations have been detected in some cases. Mutations in ACTN4, encoding alpha-actinin-4, cause an autosomal-dominant focal segmental glomerulosclerosis. alpha-actinin-4 stabilizes the podocyte cytoskeleton structure, connecting with actin filaments. WT1 mutations, causing Wilm's tumor, have been demonstrated in some CNS patients with diffuse mesangial sclerosis. Systematic investigation of genes for CNS in Japan has never been performed. METHODS: To clarify the role of mutations in these four genes, we used polymerase chain reaction (PCR) and direct sequencing to investigate all exons and exon-intron boundaries for these genes in 13 unrelated CNS patients from regional pediatric kidney disease centers in Japan. RESULTS: A novel homozygous nonsense mutation of NPHS1, E246X in exon 7, and a novel homozygous deletion mutation of NPHS1, 2156_2163del in exon 16 were detected in one patient each. A novel homozygous nonsense mutation of NPHS2, R196X in exon 5, was found in one patient, and the same heterozygous nonsense mutation was detected in another. No ACTN4 or WT1 mutations were detected. CONCLUSION: These studies demonstrate that mutation of NPHS1 is not a major cause of CNS in Japanese patients, and that mutation of NPHS2 can be responsible for CNS in this population. PMID- 15780079 TI - Detection of mutations in the COL4A5 gene by analyzing cDNA of skin fibroblasts. AB - BACKGROUND: Alport syndrome is a progressive hereditary glomerulonephritis that is characterized by hematuria, sensorineural deafness, ocular lesions, and progressive renal failure. The majority of cases (about 85%) are caused by mutations in the COL4A5 gene on the X chromosome which encodes the type IV collagen alpha5 chain (X-linked Alport syndrome). METHODS: In this study we performed a systematic analysis of the entire coding region of COL4A5 mRNA in 31 unrelated Chinese X-linked Alport syndrome patients and four controls by using reverse transcription-polymerase chain reaction (RT-PCR) and direct sequencing methods. The mRNA analyzed was isolated from cultured skin fibroblasts of Alport syndrome patients. RESULTS: The entire sequences of mRNA of the controls corresponded exactly to the published sequence. There were 28 variants detected by analyzing mRNA of COL4A5 in 28/31 patients. Of those, a total of 25 functionally significant COL4A5 mutations was confirmed in 25/31 patients by using RT-PCR method and subsequently confirmed at genomic DNA level, which included seven different mutations described in previous reports, and 18 novel mutations. The mutation detection rate was 80.6% (25/31), which is comparable with the highest previous detection sensitivity of COL4A5 mutations in evident X linked Alport syndrome using genomic DNA. Furthermore, three splicing mutations that occurred at the cryptic splice sites and would be overlooked or simply considered as intronic sequence variations by solely analyzing genomic DNA were identified in this study. CONCLUSION: RT-PCR and direct sequencing using cultured skin fibroblasts RNA is a practical approach with high sensitivity for genetic analysis in X-linked Alport syndrome patients. PMID- 15780078 TI - Modifier genes play a significant role in the phenotypic expression of PKD1. AB - BACKGROUND: Polycystic kidney disease type 1 (PKD1) is characterized by extreme variation in the severity and progression of renal and extrarenal phenotypes. There are significant familial phenotype differences; but it is not clear if this is due to differences in PKD1 mutations, differences in genetic background, or both. METHODS: A total of 315 affected relatives (83 PKD1 families) without end stage renal disease (ESRD) were evaluated for disease markers, including renal volume, creatinine clearance, proteinuria, liver cysts, and hypertension. Of these patients, 19% progressed to ESRD within 1 to 10 years after the initial examination. Nested analysis of variance was used to investigate interfamilial and intrafamilial differences in these phenotypes. Heritability analyses were used to estimate the effect of the genetic background on phenotypic variability. The age of onset of ESRD was also analyzed with an additional 389 family members from the same PKD1 families without clinical evaluation but with data on age of onset of ESRD (or age without ESRD). RESULTS: There were significant phenotype differences between patients with the same mutation and different genetic backgrounds. The phenotypic variation between patients with different mutations and different genetic backgrounds was not significantly greater than the variation between patients with the same mutation and different genetic backgrounds. However, when the 389 family members were included, both the mutation and modifier genes had significant effects on the age of onset of ESRD. Inherited differences in genetic background were estimated to account for 18% to 59% of the phenotypic variability in PKD1 disease markers in patients prior to ESRD and in the subsequent progression to ESRD (43% heritability) in the 315 patients who were clinically evaluated. CONCLUSION: Modifier loci in the genetic background are important factors in inter- and intrafamilial variability in the phenotypic expression of PKD1. The extreme intrafamilial phenotype differences are consistent with the hypothesis that one or a few modifier genes have a major effect on the progression and severity of PKD1. PMID- 15780080 TI - Differential proteomic analysis of proteins induced by glucocorticoids in cultured murine podocytes. AB - BACKGROUND: The glomerular podocyte is the kidney cell most affected during the development of nephrotic syndrome, and mutations in podocyte proteins are responsible for a variety of inherited forms of nephrotic syndrome. Although glucocorticoids are a primary treatment for nephrotic syndrome, neither their target cell nor mechanism of action are known. In order to describe the proteome of the podocyte, and to identify podocyte proteins whose expression is altered by glucocorticoids, we performed a differential proteomic analysis of control and dexamethasone-treated cultured murine podocytes. METHODS: Podocyte proteins were separated by two-dimensional-polyacrylamide gel electrophoresis (PAGE) and identified by matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and peptide fingerprinting. Comparisons of stained two-dimensional PAGE separations were used to identify proteins whose expression was altered by treatment with the glucocorticoid dexamethasone, and these results were confirmed by quantitative Western blotting. RESULTS: A total of 106 protein spots yielded MALDI-TOF results, and 92 were identified by protein fingerprinting. Of the 88 unique proteins and four protein isoforms identified, six proteins were found whose expression was altered by dexamethasone. The proteome of cultured murine podocytes is particularly rich in actin cytoskeletal proteins and proteins involved in responses to cellular stress. The change in expression of three proteins [ciliary neurotrophic factor (CNTF), alphaB-crystallin, and heat shock protein 27 (hsp27)] was confirmed by quantitative Western blotting. CONCLUSION: Three proteins with known roles in protecting cells from injury were up-regulated by dexamethasone, demonstrating that glucocorticoids exert a direct effect on cultured podocytes resulting in changes in the expression of proteins with potential relevance to the therapeutic action of glucocorticoids in diseases such as nephrotic syndrome. PMID- 15780081 TI - Evidence for low-density lipoprotein-induced expression of connective tissue growth factor in mesangial cells. AB - BACKGROUND: Although hyperlipidemia is a risk factor for the progression of renal damage, the relationship between increased plasma lipoproteins and glomerular injury is poorly defined. Connective tissue growth factor (CTGF) is emerging as a key determinant of progressive fibrotic diseases and its expression is up regulated by diabetes. To define the mechanisms through which low-density lipoproteins (LDLs) promote glomerular injury, we evaluated whether LDL can modulate the expression of CTGF and collagen I. METHODS: The effects of LDL on CTGF and collagen I expression were carried out in rat mesangial cells. RESULTS: Treatment of mesangial cells with LDL for 24 hours produced a significant increase in the protein levels of CTGF and collagen I compared to unstimulated controls. To explore if CTGF and collagen I are downstream targets for regulation by transforming growth factor-beta (TGF-beta), mesangial cells were treated with various concentration of TGF-beta for 24 hours. TGF-beta produced a concentration dependent increase in the protein levels of CTGF and collagen I. The increase in CTGF and collagen I induced by LDL was significantly inhibited by neutralizing anti-TGF-beta antibodies. Inhibition of p38(mapk) or p42/44(mapk) activities did not affect LDL-induced TGF-beta1, CTGF, and collagen I expression, whereas inhibition of c-Jun NH2-terminal kinase (JNK) suppressed LDL-induced TGF-beta, CTGF, and collagen I expression. CONCLUSION: These findings implicate JNK pathway and TGF-beta1 as key players in LDL signaling leading to CTGF and collagen I expression in mesangial cells. The data also point to a potential mechanistic pathway through which lipoproteins may promote glomerular injury. PMID- 15780082 TI - Plasminogen activator inhibitor-1 deficiency retards diabetic nephropathy. AB - BACKGROUND: Plasminogen activator inhibitor-1 (PAI-1) is increased in kidneys of humans and animals with diabetic nephropathy and is associated with extracellular matrix (ECM) accumulation. PAI-1 may promote ECM buildup by preventing plasmin and matrix metalloproteinase (MMP) activation. However, the importance and mechanism of PAI-1 action in the pathogenesis of diabetic nephropathy is unknown. METHODS: We investigated the effect of streptozotocin (STZ)-induced diabetes in wild-type (PAI-1(+/+)) mice and mice null for PAI-1 (PAI-1(-/-)). After 1 month of diabetes, animals were placed in metabolic cages for 24-hour urine collection. Total RNA was isolated from kidney cortex for reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis, and Western blots were quantitated from cortical protein. Primary mesangial cells were grown from Sprague-Dawley rats and used in signal transduction studies. RESULTS: Urinary albumin excretion (UAE) in diabetic PAI-1(+/+) mice increased >threefold, but remained unchanged in PAI-1(-/-) mice. Transforming growth factor-beta (TGF-beta) and fibronectin message and protein levels were lower in diabetic PAI-1(-/-) vs. PAI-1(+/+) mice, suggesting that PAI-1 deficiency impaired TGF-beta expression despite diabetes. Indeed, recombinant PAI-1 directly stimulated TGF-beta message and protein via mitogen-activated protein kinase (MAPK) signal transduction in cultured mesangial cells. Urokinase plasminogen activator (uPA) inhibited this PAI-1 action in a dose-dependent manner. The inhibitory effect of antibody to uPA receptor (uPAR) on PAI-1-induced TGF-beta function suggested that uPAR mediated the cellular effect of PAI-1. CONCLUSION: PAI-1 can regulate TGF-beta expression by binding to uPAR and activating the extracellular-regulated signal kinase (ERK)/MAPK pathway. Therefore, PAI-1 contributes to diabetic nephropathy by regulating TGF-beta and renal ECM production and may be a therapeutic target in diabetic nephropathy. PMID- 15780083 TI - Insulin-like growth factors inhibit podocyte apoptosis through the PI3 kinase pathway. AB - BACKGROUND: Abnormal podocyte development and progressive podocyte injury have been implicated in a number of human kidney diseases. Factors necessary for regulating development and maintenance of this cell type are only beginning to emerge. METHODS: To study the role of the insulin-like growth factor (IGF) system in regulating podocyte survival, we induced human fetal podocytes to undergo apoptosis. We demonstrated a significant increase in apoptosis when these cells were incubated in the presence of etoposide, as measured by DNA fragmentation and nuclear membrane condensation and blebbing. RESULTS: Podocyte apoptosis was reduced to control levels when the cells were coincubated in the presence of IGF 1. We showed that the protective effect of IGFs in this cell type was mediated through the activation of the phosphatidylinositol 3'-kinase (PI3K) pathway. IGF 1 stimulation resulted in the formation of the insulin receptor substrate (IRS)-1 p85 complex, an increase in PI3 kinase activity, and activation of protein kinase B (AKT/PKB) and the bcl-2 family member bad. Incubation of the podocytes with inhibitors of the PI3 kinase pathway resulted in a loss of this IGF-1 protective effect. CONCLUSION: These data demonstrate an important role for the IGF system in fetal podocyte survival in vitro, and suggest potential mediators to slow or alleviate the loss or damage of the podocyte in progressive renal disease. PMID- 15780084 TI - Thrombin stimulates proinflammatory and proliferative responses in primary cultures of human proximal tubule cells. AB - BACKGROUND: Fibrin deposition is frequently observed within the tubulointerstitium in various forms of chronic renal disease. This suggests the presence of active components of the coagulation pathway, which may contribute to the progressive deterioration in renal function. The aim of this study was to investigate the proinflammatory and fibroproliferative effects of the coagulation protease thrombin on human proximal tubular cells (PTC) in culture. METHODS: Primary cultures of PTC were established from normal kidney tissue and grown under serum-free conditions with or without thrombin or the protease-activated receptor (PAR) activating peptides TFLLRN-NH(2), SLIGKV-NH(2), and SFLLRN-NH(2) (100 to 400 micromol/L). DNA synthesis (thymidine incorporation), intracellular Ca(2+) mobilization (fura-2 fluorimetry), fibronectin secretion [enzyme-linked immunosorbent assay (ELISA), immunoblotting], monocyte chemoattractant protein-1 (MCP-1) secretion (ELISA), and transforming growth factor-beta1 (TGF-beta1) secretion (ELISA) were measured. Reverse transcription-polymerase chain reaction (RT-PCR) was used to assess PAR mRNA expression in these cells. RESULTS: Thrombin enhanced DNA synthesis, fibronectin secretion, MCP-1 secretion, and TGF-beta1 secretion in a concentration-dependent manner. Cell injury [lactate dehydrogenase (LDH) release] and cellular protein levels were unaffected. RT-PCR showed that cultures of PTC expressed mRNA transcripts for the thrombin receptors PAR-1 and PAR-3, but not PAR-4. Thrombin and each of the PAR activating peptides enhanced intracellular calcium mobilization. However, the other effects of thrombin were only fully reproduced by the PAR-2-specific peptide, SLIGKV-NH(2), only partially by SFLLRN-NH(2), (a PAR-1 peptide that can activate PAR-2), and not at all by the PAR-1-specific peptide, TFLLRN-NH(2). Thrombin-induced DNA synthesis, fibronectin, and MCP-1 secretion were unaffected by a TGF-beta neutralizing antibody, the matrix metalloproteinase (MMP) inhibitor, GM6001 and the epidermal growth factor (EGF) receptor kinase inhibitor AG1478. CONCLUSION: Thrombin initiates both proinflammatory and fibroproliferative responses in human PTC. These responses which are dependent on its protease activity appear not to be mediated by PAR-1 activation, the autocrine action of thrombin-induced TGF-beta1 secretion, MMP activation, or EGF receptor transactivation. The proinflammatory and fibroproliferative actions of thrombin on human PTC may help explain the extent of tubulointerstitial fibrosis observed in kidney diseases where fibrin deposition is evident. PMID- 15780085 TI - Midkine antisense oligodeoxyribonucleotide inhibits renal damage induced by ischemic reperfusion. AB - BACKGROUND: Midkine, a heparin-binding growth factor, is involved in the migration of inflammatory cells. The inflammatory cell migration to the tubulointerstitium of the kidney after ischemia/reperfusion (I/R) injury is attenuated in midkine gene-deficient mice, resulting in better preservation of the tubulointerstitium compared with wild-type mice. In the present investigation, we planned to evaluate the usefulness of antisense midkine for the therapy of ischemic renal failure. METHODS: Midkine antisense phosphorothioate oligodeoxyribonucleotide (ODN) at a dose of 1 mg/kg in saline was intravenously administered to mice 1 day before or after I/R. The kidneys were removed for examination 1, 2, 3, and 7 days after I/R. RESULTS: It was rapidly incorporated into proximal tubular epithelial cells, and inhibited midkine synthesis, leading to reduced migration of inflammatory cells to the injured epithelial layer. Consequently, the midkine antisense ODN-treated animals exhibited less severe renal damage than untreated or midkine sense ODN-treated animals 2 days after I/R as assessed by morphologic criteria and blood urea nitrogen (BUN) and serum creatinine levels. Midkine expression, BUN, and serum creatinine levels were not significantly different between injection of midkine antisense ODN before and after ischemic injury. CONCLUSION: These results indicate that intravenous injection of midkine antisense ODN is a candidate for a novel therapeutic strategy against acute tubulointerstitial injury induced by I/R injury. PMID- 15780086 TI - CD40 ligand (CD154) takes part in regulation of the transition to mononuclear cell dominance during peritonitis. AB - BACKGROUND: CD40 is a member of the tumor necrosis factor (TNF) family of receptors whose ligand (CD154) is found mainly on membranes of activated mononuclear cells. CD154-CD40 cross-linking is a central event in antigen presentation, B-cell activation, and regulation of cytokine and chemokine secretion from various types of cells. We have previously demonstrated in vitro the presence of CD40 on human peritoneal mesothelial cells (PMC) and have also shown that CD40 ligation synergizes with interferon-gamma (IFN-gamma) to up regulate CC chemokine secretion from these cells. The aim of the present study was to investigate the role of CD40 ligation in leukocyte recruitment during peritonitis. METHODS: Peritonitis was induced in mice by bacterial inoculation, CD40 levels were analyzed on PMC by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. CD154 levels on leukocytes were analyzed by flow cytometry and RT-PCR. Chemokines mRNA levels were analyzed by RT PCR. CD154 was blocked in vivo using monoclonal antibodies. Results. In mice inoculated by Staphylococcus epidermidis or Escherichia coli, CD40 in PMC increased twofold at 24 hours and CD154 was induced and reached a peak at 48 hours. In both Gram-positive and Gram-negative-peritonitis, peritoneal macrophages were the main peritoneal leukocyte population to express CD154. Similar results were observed in human subjects during peritonitis. Injection of CD154 blocking monoclonal antibody (MR1) reduced the mononuclear infiltrate by 50% and had no effect on granulocyte recruitment 48 hours after inoculation of S. epidermidis. CONCLUSION: Our data suggest that CD40 plays a significant role in the process of the mononuclear infiltration during peritonitis. PMID- 15780087 TI - The lipopolysaccharide-triggered mesangial transcriptome: Evaluating the role of interferon regulatory factor-1. AB - BACKGROUND: Presently, we do not have a clear picture of how the mesangial transcriptome evolves following stimulation. The present study was designed to address this, using an innate trigger to stimulate murine mesangial cells. METHODS: Three independent mesangial cell lines derived from C57BL/6 mice were stimulated with lipopolysaccharide (LPS). The mesangial cell transcriptomes were defined 1, 6, 24, and 60 hours poststimulation with LPS, using a 17,000 gene oligonucleotide array. RESULTS: Interferon regulatory factor-1 (IRF-1), ScyA2/MCP1, ScyA20/MIP3alpha (ScyB1/Gro1, and ScyB2/MIP2alpha/Gro2 were the earliest genes to be hyperexpressed after LPS stimulation. Later-appearing genes included ScyA7/MCP3, ScyD1/fractalkine, GM-CSF/CSF-2, PDGF, epiregulin, NfKb, C/EBP, TIMP-1, MMP11, MMP13, PTGS2/COX2, SpI2-1, Spp1, PAI-1, VCAM-1, C3, and defensin-beta1, among others. Several of these changes were validated by real time polymerase chain reaction (PCR) or enzyme-linked immunosorbent assay (ELISA). Rapid IRF-1 hyperexpression was also noted following stimulation of mesangial cells with peptidoglycan, poly I:poly C, interferon-gamma?(IFN-gamma), and heat-aggregated IgG. However, the blocking of IRF-1 using RNA interference and the use of mesangial cells isolated from IRF-1-deficient mice could not substantiate an obligatory role for IRF-1 in LPS-induced mesangial cell activation. Likewise, IRF-1 deficiency did not impact the development of anti glomerular basement membrane (GBM)-induced immune nephritis. CONCLUSION: Innate stimuli such as LPS appear to trigger successive waves of mesangial cell gene expression. Although IRF-1 surfaces as an "early-on, early-off" transcription factor following several different triggers, it does not appear to be an essential molecule for mesangial cell activation by innate triggers or for anti GBM disease. PMID- 15780088 TI - R-roscovitine (CYC202) alleviates renal cell proliferation in nephritis without aggravating podocyte injury. AB - BACKGROUND: Cyclin-dependent kinase (CDK) inhibition is a new therapeutic approach to proliferative glomerulonephritides. CDK2 is required for G(1)/S transition and DNA synthesis and is inhibited by CYC202 (R-roscovitine). Since podocytes express CDK2 in nephritis and since loss of podocytes contributes to glomerulosclerosis, the rationale of the present study was to test whether CDK2 inhibition is safe in instances of podocyte injury. METHODS: Rats with passive Heymann nephritis, a model of membranous glomerulonephritis, were treated (day 3 to 30) with vehicle, low (25 mg/kg/day), or high (50 mg/kg/day) doses of CYC202. RESULTS: On day 27, blood pressure was normal in nephritic controls and was dose dependently reduced by CYC202. Urinary albumin excretion did not differ between the groups on days 9, 16, 23, and 30. To investigate podocyte injury, we assessed the glomerular de novo expression of desmin, which was markedly up-regulated in almost all passive Heymann nephritis glomeruli but was not significantly different between the three groups. No tubulointerstitial de novo expression of desmin or alpha-smooth muscle actin (alpha-SMA), or tubulointerstitial monocyte/macrophage infiltration was noted in any group. Biologic activity of CYC202 was evident in the form of a dose-dependent decrease in the number of glomerular and tubulointerstitial mitotic figures as compared to vehicle alone. Glomerular immunostaining for cyclin D1, a marker for G(0) to G(1) transition, was significantly decreased in CYC202 treated groups at day 9. CONCLUSION: Whereas inhibition of CDKs by CYC202 reduced intrarenal cell proliferation in passive Heymann nephritis it did not aggravate podocyte damage, suggesting that this novel therapeutic approach is safe in renal diseases characterized by podocyte injury. PMID- 15780089 TI - Mechanisms of antioxidant and pro-oxidant effects of alpha-lipoic acid in the diabetic and nondiabetic kidney. AB - BACKGROUND: alpha-Lipoic acid is a potent antioxidant that improves renal function in diabetes by lowering glycemia, however, the mechanisms by which alpha lipoic acid exerts its antioxidant effects are not completely understood. METHODS: Metabolic parameters, renal function, and morphology, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity and subunit expression were analyzed in nondiabetic and streptozotocin-induced diabetic rats fed normal rat chow (control) with or without alpha-lipoic acid (30 mg/kg body weight) for 12 weeks. RESULTS: Blood glucose was increased with diabetes (nondiabetic + control 89 +/- 3 mg/dL and diabetic + control 336 +/- 28 mg/dL) and was similar with alpha-lipoic acid treatment (diabetic +alpha-lipoic acid 351 +/- 14 mg/dL). In contrast, alpha-lipoic acid attenuated albuminuria (nondiabetic + control 8.9 +/- 1.3 mg/day; diabetic + control 28.1 +/- 4.6 mg/day; and diabetic +alpha lipoic acid 17.8 +/- 1.2 mg/day) associated with diabetes. Similarly, alpha lipoic acid attenuated glomerulosclerosis (nondiabetic + control 0.22 +/- 0.01; diabetic + control 0.55 +/- 0.04; diabetic +alpha-lipoic acid 0.36 +/- 0.03), tubulointerstitial fibrosis (nondiabetic + control 0.42 +/- 0.18; diabetic + control 1.52 +/- 0.05; diabetic +alpha-lipoic acid 1.10 +/- 0.05), superoxide anion (O(.-) (2)) generation (nondiabetic +control 15.8 +/- 1.7; diabetic +control 87.1 +/- 3.5; diabetic +alpha-lipoic acid 25.5 +/- 3.3 RLU/mg protein), and urine 8-isoprostane (8-iso) excretion (nondiabetic + control 7.4 +/- 1.4; diabetic + control 26.0 +/- 4.5; diabetic +alpha-lipoic acid 19.6 +/- 5.6 ng/day) associated with diabetes. alpha-Lipoic acid also reduced kidney expression of NADPH oxidase subunits p22phox and p47phox. Surprisingly, alpha-lipoic acid appears to cause pro-oxidant effects in nondiabetic animals, resulting in increased albuminuria (nondiabetic +alpha-lipoic acid 14.2 +/- 1.2 mg/day), increase in plasma creatinine levels (nondiabetic + control 59 +/- 6; diabetic + control 68 +/- 6; nondiabetic +alpha-lipoic acid 86 +/- 9; diabetic +alpha-lipoic acid 69 +/- 7 mumol/L), exacerbated glomerulosclerosis and tubulointerstitial fibrosis, increased O(.-) (2) generation, up-regulated p22phox and p47phox expression and increased 8-iso excretion. CONCLUSION: We conclude that alpha lipoic acid improves albuminuria and pathology in diabetes by reducing oxidative stress, while in healthy animals, alpha-lipoic acid may act as a pro-oxidant, contributing to renal dysfunction. PMID- 15780091 TI - Positive correlation between sialyl Lewis X expression and pathologic findings in renal cell carcinoma. AB - BACKGROUND: Interaction between tumor cells and endothelium plays a major role in cancer invasion and metastasis. Among various cell adhesion molecules, the cognate interaction between sialyl Lewis antigen expressed in the tumor cell surface and E-selectin expressed on endothelial cells is considered to be crucial for the tumor cell adhesion to the endothelium. METHODS: The sialyl Lewis X (sL(X)) expression in 45 specimens from renal cell carcinoma patients was examined using immunohistochemistry. RESULTS: In this study, we demonstrate that the immunoreactivity for sL(X) in renal cell carcinoma specimens not only correlates with conventional histopathologic parameters but also serves as a useful indicator for the prognosis of renal cell carcinoma. CONCLUSION: Since beneficial effect of cimetidine has been reported and ascribed to its inhibitory action on the expression of E-selectin, a ligand molecule of sialyl Lewis antigen, cimetidine may also show inhibitory effect on the tumor recurrence and metastasis of renal cell carcinoma with high level of sL(X) expression. PMID- 15780090 TI - HIV-associated immune complex glomerulonephritis with "lupus-like" features: a clinicopathologic study of 14 cases. AB - BACKGROUND: While the most common glomerular lesion associated with human immunodeficiency virus (HIV) infection is collapsing focal segmental glomerulosclerosis (FSGS) [HIV-associated nephropathy (HIVAN)], immune complex mediated forms of glomerulonephritis have been increasingly reported. One form of glomerulonephritis that has been described in the HIV-infected population is immune complex glomerulonephritis with "lupus-like" features, characterized by histologic, immunohistologic, and ultrastructural features resembling lupus nephritis, but occurring in patients without evidence of systemic lupus erythematosus (SLE). Data regarding clinical outcomes in patients with this form of glomerulonephritis are very limited. METHODS: We reviewed pathology reports for all native renal biopsy specimens from HIV-positive patients processed at our center from January 1999 through December 2003. Of 77 total specimens, 14 met the following criteria for lupus-like glomerulonephritis: (1) immunofluorescence microscopy showed granular glomerular staining for IgG, IgA, IgM, C3 and C1q, with > or=1+ (0 to 4+ scale) staining for C1q; and (2) the patient's serum was negative for antinuclear antibodies (ANA), or weakly positive (titer < or =1:80) for ANA and negative for antidouble-stranded DNA. RESULTS: Clinically, ten of the 14 patients with lupus-like glomerulonephritis presented with nephrotic syndrome, all had microscopic hematuria, and nine had serum creatinine >3.0 mg/dL. All but one were African American. Histologically, seven biopsies showed diffuse proliferative glomerulonephritis, six focal proliferative glomerulonephritis, and one membranous nephropathy. All but two biopsies showed moderate or severe chronic change, and three showed concurrent HIVAN. Ten of the 14 patients developed end-stage renal disease (ESRD) within 1 year of the biopsy. Nine of these ten patients presented with proteinuria >5.0 g/24 hours and nephrotic syndrome, while three of four patients who did not develop ESRD had proteinuria < or =3.0 g/24 hours. CONCLUSION: Lupus-like glomerulonephritis, defined by immunohistologic features and absence of serologic evidence of SLE, is not an uncommon form of glomerular disease in HIV-infected patients undergoing a renal biopsy. Renal outcomes in these patients were poor, although this may be due largely to most patients presenting with advanced disease. PMID- 15780092 TI - IL-2-mediated apoptosis of kidney tubular epithelial cells is regulated by the caspase-8 inhibitor c-FLIP. AB - BACKGROUND: Tubular epithelial cells (TECs) are essential in the maintenance of kidney function. Apoptosis of TECs occur during acute and chronic renal allograft rejection as well as other forms of renal injury, including autoimmune nephritis. The regulation of TEC apoptosis by proinflammatory cytokines associated with renal inflammation [e.g., interleukin (IL)-2 and interferon-gamma (IFN-gamma)] has not been extensively investigated. METHODS: Apoptosis in murine TECs was determined by FACS with annexin-V or ligation-mediated-polymerase chain reaction (LM-PCR) and mRNA levels by reverse transcription (RT)-PCR or Northern blot. Protein expression was observed using Western blot. RESULTS: IL-2R (CD25) was expressed by murine TECs and up-regulated by IL-2. Both IL-2 and IFN-gamma induced TEC apoptosis and activated caspase-8. Apoptosis with IL-2 was concentration-dependent and blocked by z-IETD-fmk, a specific caspase-8 inhibitor. Apoptosis with IFN-gamma was associated with increased surface expression of Fas, while IL-2 had no effect on Fas. IL-2 did not induce apoptosis in Fas-deficient TECs (M3.1-lpr) suggesting IL-2 regulation of caspase-8 activity requires Fas. Consistent with this, IL-2 but not IFN-gamma was found to decrease mRNA and protein expression of c-FLIP, an endogenous caspase-8 inhibitor in murine TECs. Overexpression of c-FLIP in TECs (CS3.7-FLIP) blocked apoptosis and caspase-8 activation with both IFN-gamma and IL-2. c-FLIP expression was found in kidney cortex, primary and cloned TECs, suggesting c-FLIP is likely a key regulator of caspase-8-mediated apoptosis in vivo. CONCLUSION: This is the first report of c-FLIP regulation by IL-2 in renal TECs. Augmentation of c-FLIP in TECs may enhance an endogenous mechanism by which TECs normally resist injury to caspase-8-mediated apoptosis and thus may be a useful and novel strategy to prevent tubular injury in transplant rejection and autoimmune nephritis. PMID- 15780094 TI - Guanylin and uroguanylin regulate electrolyte transport in isolated human cortical collecting ducts. AB - BACKGROUND: Guanylin and uroguanylin link intestinal and renal electrolyte and water transport. Their function in intestine is well studied, but renal actions are less understood. Uroguanylin concentrations are increased in patients with chronic renal failure, nephrotic syndrome, or those on dialysis. Guanylate cyclase C (GC-C) is the receptor first described for these peptides. In guanylate cyclase C-deficient mice guanylin- and uroguanylin-induced renal natriuresis, kaliuresis, and diuresis are retained. METHODS: Effects of guanylin and uroguanylin on principal cells of human cortical collecting ducts (CCD) isolated from kidneys after tumor nephrectomy were investigated. Reverse transcription polymerase chain reaction (RT-PCR), slow whole-cell patch-clamp, and microfluorimetric analysis of intracellular Ca(2+) were used. Here we present first functional measurements of isolated human CCD. RESULTS: Principal cells of CCD were identified by the amiloride-induced hyperpolarization of principal cells (-3.8 +/- 0.3 mV) (N= 52). Cells depolarized upon guanylin or uroguanylin (each 10 nmol/L) by 3.3 +/- 0.8 mV (N= 12) and 3.4 +/- 0.5 mV (N= 18), respectively, but were hyperpolarized by 8Br-cyclic guanosine monophosphate (cGMP) (100 micromol/L) (-3.0 +/- 0.2 mV) (N= 4). mRNA for GC-C was not detected in CCD. Effects of both peptides were inhibited by Ba(2+) (1 mmol/L) or phospholipase A(2) (PLA(2)) inhibition (AACOCF(3)) (5 micromol/L). CONCLUSION: These findings suggest a new cGMP- and GC-C-independent but PLA(2)-dependent signaling pathway for these peptides in the kidney. Most likely guanylin and uroguanylin inhibit luminal K(+) channels of principal cells of human CCD via this pathway. This depolarization of principal cells consequently reduces the driving force of Na(+) and water reabsorption, explaining natriuresis and diuresis caused by these peptides. PMID- 15780093 TI - Circadian clock genes directly regulate expression of the Na(+)/H(+) exchanger NHE3 in the kidney. AB - BACKGROUND: Daily rhythms in mammalian physiology are generated by a transcription/translation feedback loop orchestrated by a set of clock genes. However, little is known about the molecular cascade from the clock gene oscillators to cellular function. METHODS: The mRNA expression profiles of NHE3 and clock genes were examined in mice and rat kidneys. First, luciferase assays followed by a site directed mutagenesis of an E-box sequence were used to assess the CLOCK:BMAL1-transactivated NHE3 promoter activity. A direct binding of CLOCK:BMAL1 heterodimers to an E-box sequences of NHE3 promoter was confirmed by electrophoretic mobility shift assay (EMSA). RESULTS: We present evidence that renal tubular NHE3, the Na(+)/H(+) exchanger critical for systemic electrolyte and acid-base homeostasis, is a clock-controlled gene regulated directly by CLOCK:BMAL1 heterodimers in kidneys. NHE3 mRNA level in rat kidney displayed circadian kinetics, and this circadian expression was severely blunted in homozygous CRY1/2 double-deficient mice, suggesting that the transcriptional machinery of peripheral clocks in renal tubular cells directly regulates the circadian expression of NHE3. By analyzing the 5' upstream region of the NHE3 gene, we found an E box critical for the transcription of NHE3 via the CLOCK:BMAL1-driven circadian oscillator. The circadian expression of NHE3 mRNA was reflected by oscillating protein levels in the proximal tubules of the rat kidney. CONCLUSION: NHE3 should represent an output gene of the peripheral oscillators in kidney, which is regulated directly by CLOCK:BMAL1 heterodimers. PMID- 15780095 TI - A biologic role of HIF-1 in the renal medulla. AB - BACKGROUND: Activation of hypoxia-inducible factor-1 (HIF-1) is the primary defensive mechanism against hypoxia. HIF-1 activation generally occurs in pathologic disruption of tissue oxygenation. However, a biologic role of HIF-1 in the medulla of the kidney, which is considered perpetually hypoxic under physiologic conditions due to its unique circulation, remains to be elucidated. METHODS: The expression of HIF-1alpha was detected by immunohistochemical analysis. Functional studies of HIF in medulla were carried out by gene transfer of various plasmids by retrograde injection via ureter. RESULTS: Our immunohistochemical analysis detected HIF-1alpha in the inner stripe and the inner medulla of normal rats. Water deprivation increased the number of HIF 1alpha-positive cells, which may be mediated by an increase in medullar workload and a decrease in local blood flow. To perform functional studies, we performed gene transfer. Efficient expression of the transgene was confirmed using an enhanced green fluorescent protein (E-GFP) expressing vector. Our histologic and immunoblotting analysis detected the transgene product at the inner medulla and the inner stripe 48 hours after injection. Administration of negative-dominant HIF induced severe damage in the medulla of normal rats. In contrast, gene transfer of constitutively active HIF (HIF/VP16) induced expression of various HIF-regulated genes and protected the medulla against ischemic insults. CONCLUSION: Our studies demonstrated a crucial role of HIF in the renal medulla under normal and hypoxic circumstances. PMID- 15780096 TI - Development of diabetic nephropathy in the Milan normotensive strain, but not in the Milan hypertensive strain: possible permissive role of hemodynamics. AB - BACKGROUND: Rats of the Milan normotensive strain develop spontaneous glomerulosclerosis, whereas those of the Milan hypertensive strain are resistant to renal disease, possibly due to intrarenal artery hypertrophy protecting from systemic hypertension. To assess the role of hemodynamic versus metabolic factors in diabetic nephropathy, we investigated whether streptozotocin-induced diabetes accelerates glomerulosclerosis in Milan normotensive and/or removes (the hemodynamic) protection in Milan hypertensive rats by reducing preglomerular vascular resistance. METHODS: Diabetic and nondiabetic Milan normotensive, hypertensive, and progenitor Wistar rats were followed for 6 months for the assessment of renal function and structure. RESULTS: Proteinuria increased in nondiabetic and diabetic normotensive and, to a lesser extent, in diabetic Wistar, but not hypertensive rats. Serum creatinine increased and creatinine clearance decreased in nondiabetic and diabetic normotensive rats at 6 months. At 1.5 months, diabetic normotensive, but not hypertensive rats showed increased glomerular filtration rate and filtration fraction, suggesting glomerular hypertension. Diabetic nephropathy was detected in diabetic normotensive and Wistar, but not hypertensive rats. Glomerular extracellular matrix and TGF-beta mRNA levels increased with diabetes (and age) in normotensive, but not hypertensive rats. Arterioles and interlobular arteries showed increased media thickness in hypertensive versus normotensive rats, with diabetes reducing it only in the normotensive. CONCLUSION: These data show that Milan hypertensive rats are not susceptible to diabetic nephropathy, at variance with glomerulosclerosis-prone Milan normotensive rats, thus indicating the importance of genetic background. Our study suggests that the nature of this (genetic) protection might be hemodynamic, with intrarenal artery hypertrophy preventing diabetes-induced loss of autoregulation. PMID- 15780098 TI - Hypertension promotes integrin expression and reactive oxygen species generation by circulating leukocytes. AB - BACKGROUND: Compelling evidence has emerged pointing to the role of oxidative stress in the pathogenesis of hypertension (HTN) in experimental animals. Excess production of reactive oxygen species (ROS) in the renal and vascular tissues has been shown to raise arterial pressure via inactivation of nitric oxide and generation of isoprostanes. Accumulation of inflammatory cells in the renal interstitium has been shown to increase ROS generation in the kidneys of hypertensive animals. In addition, considerable evidence has emerged pointing to spontaneous activation of circulating leukocytes in animals and humans with hereditary HTN. This study was designed to explore whether induction of HTN in genetically normal animals can lead to spontaneous activation and ROS production in circulating leukocytes. METHODS: Integrin expression, superoxide, and hydrogen peroxide production were assessed by flow cytometry in the circulating and splenic leukocytes of Sprague-Dawley rats rendered hypertensive by abdominal aorta banding above the renal arteries, and in sham-operated control rats. RESULTS: The hypertensive animals studied 4 weeks after abdominal aorta banding exhibited a significant increase in superoxide and H(2)O(2) production in the circulating granulocyte, and a marked increase in H(2)O(2) production in the blood and splenic helper and cytotoxic T-lymphocytes. This was coupled with a significant up-regulation of CD18 and CD11a in splenic helper T-cells and cytotoxic T cells, and of CD18 in the circulating helper T cells. CONCLUSION: Induction of HTN in genetically normotensive rats causes a spontaneous increase in ROS generation in the circulating and splenic leukocytes. This phenomenon can contribute to systemic oxidative stress, inflammation, cardiovascular and renal complications in hypertensive animals. PMID- 15780097 TI - Effects of angiotensin-(1-7) blockade on renal function in rats with enhanced intrarenal Ang II activity. AB - BACKGROUND: Increasing evidence suggests that angiotensin-(1-7) [Ang-(1-7)] acts as an endogenous antagonist of Ang II when the renin-angiotensin system (RAS) is activated. In the present study, we therefore compared the effects of acute intrarenal (i.r.) Ang-(1-7) receptor blockade on renal function under conditions of normal and increased intrarenal Ang II concentration. METHODS: Salt-replete Hannover-Sprague Dawley rats (HanSD) served as control animals. As models with enhanced action of Ang II we first used transgenic rats harboring the Ren-2 renin gene (TGR), second, Ang II-infused rats, third, 2-kidney, 1-clip (2K1C) hypertensive rats on normal salt intake, and fourth, salt-depleted TGR and HanSD. RESULTS: I.r. Ang-(1-7) receptor blockade elicited significant decreases in glomerular filtration rate (GFR), renal plasma flow (RPF), and sodium excretion in 2K1C rats, and in salt-depleted TGR and HanSD. In contrast, i.r. Ang-(1-7) receptor blockade did not significantly change GFR, RPF, and sodium excretion in salt-replete TGR and HanSD, or in Ang II-infused rats. CONCLUSION: These findings suggest that under conditions of normal intrarenal RAS activity and increased intrarenal Ang II action by infusion of Ang II or by insertion of a renin gene in salt-replete conditions, Ang-(1-7) is not an important factor in the regulation of renal function. In contrast, under conditions of endogenous RAS activation due to clipping of the renal artery or to sodium restriction, Ang-(1-7) serves as opponent of the vasoconstrictor actions of Ang II. PMID- 15780099 TI - Use of sirolimus and low-dose calcineurin inhibitor in lung transplant recipients with renal impairment: results of a controlled pilot study. AB - BACKGROUND: Renal failure induced by calcineurin-inhibitor agents is a common complication of lung transplantation. Sirolimus, a macrolide immunosuppressant with a distinct mechanism of action, may prevent renal failure but was found to have a high infectious and toxicity rate in the only relevant study conducted so far. The aim of the present prospective pilot study was to assess the benefit of sirolimus combined with low-dose calcineurin inhibitors in this patient population. METHODS: Sixteen lung transplant recipients with post-transplantation renal dysfunction were allocated to receive the standard immunosuppression regimen or a combination sirolimus/low-dose calcineurin-inhibitor regimen. Target trough levels of sirolimus were 4 to 8 ng/mL. Tacrolimus was tapered down to target trough levels of 4 to 8 ng/mL and cyclosporine to 80 to 120 ng/mL. Duration of follow-up was 18 months. RESULTS: At the end of follow-up, the sirolimus group showed a significant improvement in creatinine clearance (42.6 mL/min vs. 32.5 mL/min, P= 0.05), whereas the control group showed a significant reduction (32.3 mL/min vs. 40.3 mL/min, P= 0.02). The difference between the groups was statistically significant (P < 0.0001). Acute rejection episodes occurred in 2 patients in the sirolimus group and 1 patient in the control group (P= NS). Pneumonia developed in 6 study patients and 4 controls; all responded to antibiotics. CONCLUSION: Sirolimus combined with low-dose calcineurin inhibitors appears to be a safe and effective alternative immunosuppressive therapy to sirolimus alone in lung transplant recipients with renal failure. Graft function is preserved, and infection and drug toxicity rates are low. PMID- 15780100 TI - Microalbuminuria three years after recovery from Escherichia coli O157 hemolytic uremic syndrome due to municipal water contamination. AB - BACKGROUND: Knowledge of the long-term renal prognosis of diarrhea associated hemolytic uremic syndrome (HUS) is important for patient counseling and follow up. However, estimates of long-term risk are highly variable, with previous studies not using a healthy control group. METHODS: A municipal water system in the small rural town of Walkerton, Ontario, became contaminated with Escherichia coli O157:H7 in 2000. A cohort of 19 children who recovered from HUS was randomly age- and sex-matched to 38 children with no symptoms at the time of the outbreak. Both groups had detailed renal function testing 3 years after the outbreak, including a random urine albumin to creatinine, glomerular filtration rate estimated by Schwartz formula, and automated and manual blood pressure measurements. RESULTS: There were no baseline differences between the groups with respect to age (mean 4.8 years, range 1 to 15), sex, or birth weight (mean 3.4 kg). In follow-up there were no differences between the groups in body surface area (mean 1.0 m(2)), or in the methods by which renal function was assessed. Compared to the group with no symptoms, patients with HUS demonstrated more microalbuminuria [32% vs. 5%, relative risk 4.8 (95% CI 1.1 to 22.0)], a nonsignificant trend toward lower GFR (124 vs. 134 mL/min per 1.73 m(2)), and no difference in blood pressure. CONCLUSION: Children may demonstrate microalbuminuria 3 years after recovering from HUS. Longer follow-up is needed to determine if this finding has clinical relevance and utility. PMID- 15780101 TI - Higher prevalence of anemia with diabetes mellitus in moderate kidney insufficiency: The Kidney Early Evaluation Program. AB - BACKGROUND: The Kidney Early Evaluation Program (KEEP 2.0) cross-sectional, community-based study, targeted individuals at increased risk for kidney disease and measured blood glucose, creatinine, and hemoglobin. METHODS: KEEP 2.0 screening data were used to determine the prevalence of anemia by level of kidney function and diabetes status. Estimated glomerular filtration rate (EGFR) was calculated using serum creatinine values, and categorized as > or =90, 60-89, 30 59 and <30 mL/min/1.73 m(2). Anemia was defined as hemoglobin <12 g/dL in men and in women aged >50 years, and <11 g/dL in women < or =50 years. Diabetes was defined as participant-reported diagnosis, fasting glucose >125 mg/dL, or nonfasting glucose >200 mg/dL. RESULTS: Data were available on 5380 participants screened from August 2000 through December 2001. Diabetes was present in 26.9% of participants, and anemia in 7.7%; 15.9% of participants had at least moderately reduced kidney function (EGFR <60 mL/min/1.73 m(2)). In participants with diabetes, anemia prevalence at the 4 levels of descending EGFR were 8.7%, 7.5%, 22.2%, and 52.4%, compared with 6.9%, 5.0%, 7.9%, and 50.0% in persons without diabetes. In a multivariable model, participants of non-white race/ethnicity, those with diabetes and those with EGFR <30 or 30-59 mL/min/1.73 m(2) had significantly increased odds of anemia. In addition, a significant sex-diabetes interaction was identified; odds of anemia were 4-fold greater in men than women with diabetes relative to sex-matched participants without diabetes. CONCLUSION: Diabetes was independently correlated with anemia, more so in men than women, and may be linked to premature expression of anemia in persons with moderate reductions in kidney function. PMID- 15780102 TI - Renal replacement therapy for diabetic end-stage renal disease: data from 10 registries in Europe (1991-2000). AB - BACKGROUND: There is concern about the rising prevalence of type 2 diabetes mellitus and of the resultant nephropathy. This study uses data from the European Renal Association-European Dialysis and Transplant Association (ERA-EDTA) Registry to provide information on the epidemiology and outcome of renal replacement therapy (RRT) for end-stage renal disease (ESRD) due to diabetic nephropathy (DN). METHODS: Data from the following 10 registries: Austria, French speaking Belgium, Denmark, Finland, Greece, Norway, Scotland (UK), Catalonia (Spain), Sweden, and The Netherlands were combined. Average annual changes (%) were estimated by Poisson regression. Analyses of mortality were performed by Cox regression. RESULTS: An increase in patients with type 2 DN entering RRT has been observed (+11.9% annually, P < 0.05), while large differences in RRT incidence in this disease continue to exist between countries in Europe. There was a reduction in mortality during the first 2 years on dialysis therapy among patients with type 2 DN (AHR 0.96, 95%CI 0.94-0.97 annually). The mortality among transplant recipients decreased for both type 1 DN and nondiabetic ESRD (non DN) within the 1995-1998 cohort (type 1 DN: AHR 0.49, 95% CI 0.35-0.68; non DN: AHR 0.79, 95% CI 0.69-0.90) compared to the 1991-1994 cohort. CONCLUSION: This report has shown that during the last decade there has been a marked increase in the incidence of RRT for type 2 DN. Survival analysis showed that over the period 1991-1999 the mortality rates of all dialysis patients and of type 1 diabetic and nondiabetic renal transplant recipients have fallen. PMID- 15780103 TI - A systematic review of the effect of nocturnal hemodialysis on blood pressure, left ventricular hypertrophy, anemia, mineral metabolism, and health-related quality of life. AB - BACKGROUND: Nocturnal hemodialysis is a novel form of dialysis where patients perform dialysis 6 nights per week while they sleep. Multiple publications report significant improvements in selected clinical outcomes, although the strength of these results is limited by shortcomings in study design. A systematic review of the current available literature was undertaken to examine the effect of nocturnal hemodialysis on key health outcomes. METHODS: An inclusive search of medical databases was undertaken to identify all nocturnal hemodialysis studies. These results were manually reviewed for relevance to nocturnal hemodialysis and its impact on the following predefined health outcomes: blood pressure control, left ventricular hypertrophy, anemia, mineral metabolism, and health related quality of life. Case reports, short-term studies (<4 weeks), studies without comparator groups, and studies not reporting data in a quantitative fashion were excluded. The results of the remaining studies were reported in tabular format. RESULTS: Of the initial 270 studies identified, only 14 met inclusion/exclusion criteria. No studies examining the impact of nocturnal hemodialysis on mortality were identified. All studies reported improved blood pressure control after conversion to nocturnal hemodialysis. Data regarding the other health outcomes of interest revealed mixed results. CONCLUSION: Nocturnal hemodialysis is a potential alternative to conventional intermittent hemodialysis. Before significant resources are invested in initiating nocturnal hemodialysis programs, further data on mortality and cardiovascular morbidity, preferably from randomized clinical trials, are required. PMID- 15780104 TI - The clinical characteristics of HIV-infected patients receiving dialysis in France between 1997 and 2002. AB - BACKGROUND: In 1997, 0.38% of dialysis patients in France were infected by human immunodeficiency virus (HIV). No prevalence data were available in France since the widespread introduction of highly active antiretroviral therapy. METHODS: This was a cross-sectional epidemiologic survey. A questionnaire was sent to all French dialysis centers in July 2002. The centers that did not respond were sent 3 additional mailing reminders. Finally, the nonresponding centers were called early in 2004. RESULTS: Of the 27,577 patients on hemodialysis and 587 patients on peritoneal dialysis, 190 patients (0.67%) were infected by HIV. HIV-associated nephropathy was the cause of renal failure in 39.8% patients. Mean age was 44.6 +/- 10.9 years, the mean duration of dialysis was 4.9 +/- 5.9 years, the mean known duration of HIV infection was 8.9 +/- 5.6 years. Eighty-two percent of patients received antiretroviral therapy (ART). Fifty-eight percent of ART treated patients had an undetectable HIV plasma viral load with a median CD4+ T cell count 303/mm(3). CONCLUSION: The prevalence of HIV infection among French dialysis patients was 0.67% in late 2002, a 79% increase since 1997. Possible reasons for this large increase include increased access to dialysis, better general status of HIV dialysis patients, and increasing proportion of patients originating from Africa and the Caribbean. The current efficacy of ART makes renal transplantation a realistic option for these young patients. PMID- 15780105 TI - Correlation of renal histopathology with sonographic findings. AB - BACKGROUND: Judgments about irreversible renal disease are frequently based on the sonographic appearance of the kidneys. However, the sensitivity and specificity of sonography in identifying chronic, irreversible disease have never been determined, and the specific pathologic changes that increase renal cortical echogenicity have not been defined. METHODS: We retrospectively compared sonographic parameters (length, quantitative echogenicity, cortical thickness, and parenchymal thickness) to biopsy findings of glomerular sclerosis, tubular atrophy, interstitial fibrosis, and interstitial inflammation in 207 patients. RESULTS: Echogenicity showed the strongest correlation with all 4 histologic parameters (r= 0.28-0.35). Renal size was significantly correlated with glomerular sclerosis (r=-0.26) and tubular atrophy (r= 0.20). Parenchymal thickness, but not cortical thickness, correlated with tubular atrophy (r=-0.23). By multivariate analysis, tubular atrophy and interstitial inflammation, but not interstitial fibrosis, were significant determinants of cortical echogenicity. Severe chronic disease (>50% sclerosed glomeruli or a score of 3 out of 5 or greater for tubular atrophy or interstitial fibrosis) was present in 69% and 47% of patients with combined renal length <20 cm and >20 cm, respectively (P= <0.05). For cortical echogenicity >1.0 (>liver echogenicity) and 1.0. CONCLUSION: Cortical echogenicity is the sonographic parameter that correlates best with renal histopathology. Although size or echogenicity alone are poor predictors of chronic irreversible disease, the likelihood of treatable disease in small kidneys with increased cortical echogenicity is very low. PMID- 15780106 TI - Autonomic function in elderly uremics studied by spectral analysis of heart rate. AB - BACKGROUND: Aging determines an altered response of the autonomic nervous system (ANS) to physiologic stresses. A widespread autonomic damage is well recognized in chronic renal failure (CRF). METHODS: We studied 30 CRF patients, aged 19 to 85 years, who were on bicarbonate hemodialysis. Surface electrocardiogram was recorded on lying and 65 degrees head-up tilt standing positions. A dedicated software, using an autoregressive modeling technique, allowed to calculate power spectral analysis (PSA) of heart rate variability, assessing a low-frequency band in the range 0.03 to 0.15 Hz, and a high-frequency band in the range 0.15 to 0.33 Hz. Low-frequency and high-frequency components are regarded, but not invariably, as specific markers of sympathetic and parasympathetic activities, respectively, and the low-frequency/high frequency ratio as an index of sympathovagal balance. RESULTS: In normal controls, low-frequency band value and low-frequency/high frequency ratio on standing resulted significantly reduced in the group older than 65 years when compared with those younger than 65 years; an opposite finding was seen in high-frequency band value on standing. In uremic patients, low frequency band on lying resulted significantly lower only in elderly uremics when compared with elderly controls, whereas low-frequency band on standing was significantly lower in elderly than in younger uremics. Regression analysis showed a significant inverse relationship between aging and most low-frequency band values, especially in uremics. The comparison of linear regression models confirmed that a sympathetic autonomic derangement is greatly present in older uremics, in particular after 50 years of age. CONCLUSION: Our data support assertion that combination of aging and CRF increases the chance of autonomic derangement being present. PMID- 15780107 TI - Incidence and etiology of acute renal failure among ambulatory HIV-infected patients. AB - BACKGROUND: Acute renal failure (ARF) is a cause of renal dysfunction in human immunodeficiency virus (HIV)-infected patients. Its incidence and causes have not been studied since the introduction of highly active antiretroviral therapy (HAART) in HIV ambulatory patients. METHODS: This is a prospective cohort study of 754 HIV patients, 18 years or older, seen at a university-based infectious disease clinic between 2000 and 2002. ARF was identified using proportional increases in serum creatinine from baseline and by chart review. Clinical conditions were assessed at the time of the ARF event. ARF incidence rates (IR) were calculated by dividing the number of events by person time at risk. To compare patients with and without ARF, t test or chi-square test were used. RESULTS: Patient's mean age was 40 years; 68% were male and 61% were black. One hundred-eleven ARF events occurred in 71 subjects (IR 5.9 per 100 person-years; 95% CI 4.9, 7.1). ARF was more common in men, in those with CD4 cell count <200 cells/mm(3), and HIV RNA levels >10,000 copies/mL. These patients more often had acquired immunodeficiency syndrome (AIDS), hepatitis C infection (HCV), and have received HAART. ARF was mainly community-acquired, due to prerenal causes or acute tubular necrosis, and associated with opportunistic infections and drugs. Liver disease was a cause of ARF in HCV-infected patients. CONCLUSION: ARF is common in ambulatory HIV patients. Immunosuppression, infection, and HCV are important conditions associated with ARF in the post-HAART era. PMID- 15780108 TI - Modeling the implications of changes in vascular calcification in patients on hemodialysis. AB - BACKGROUND: The Treat-to-Goal Study found that sevelamer slowed the progression of coronary calcification in patients on hemodialysis compared to calcium-based phosphate binders. To understand the implications of this effect for cardiovascular events, risk equations are needed. METHODS: Data on 179 patients on hemodialysis treated at one center in France included biochemical values during the year prior to study entry, patient characteristics, and cardiovascular events over an average of 4 years. As arterial calcification was evaluated ultrasonographically and quantified using a 0 to 4 score, an equation relating this to the electron-beam tomography (EBT)-based calcification score used in the trial was developed and applied to all patients. The estimated scores were then used in survival and Cox proportional hazards analyses of cardiovascular events in relation to the degree of calcification, controlling for other characteristics. RESULTS: Mean age at inclusion was 54 years, dialysis vintage 70 months, average follow-up 49 months; 32% suffered an event. The calcification score, diabetes, C-reactive protein (CRP), diastolic blood pressure, gender, smoking and hypertension are independent predictors of cardiovascular risk. The resulting equation indicates that, relative to a calcification score below 400, the risk of an initial event increases 44% for a score of 600, and more than doubles for a score of 1000. CONCLUSION: In the absence of long-term follow-up studies, these equations permit quantification of the expected long-term clinical consequences of the impact of various phosphate binders on vascular calcification. Together with resource use and cost information, these equations are key inputs for formal cost-effectiveness analyses. PMID- 15780109 TI - Homocysteine in chronic kidney disease: Effect of low protein diet and repletion with B vitamins. AB - BACKGROUND: Data are limited on the determinants of homocysteine (tHcy) and its relationship with nutritional indices, and dietary protein intake, in the earlier stages of chronic kidney disease (CKD). METHODS: Levels of tHcy were assayed at baseline (N= 804) and 1 year postrandomization (N= 678) in the Modification of Diet in Renal Disease (MDRD) Study [study A, glomerular filtration rate (GFR) 25 to 55 mL/min/1.73 m(2) and study B GFR 13 to 24 mL/min/1.73 m(2)]. Participants were randomly assigned to different blood pressure targets and protein diets and all subjects received a multivitamin supplement containing 1 mg of folic acid, 10 mg pyridoxal 5'-phosphate (PLP) and 6 mug of vitamin B(12). Multivariable analyses were used to evaluate determinants of tHcy at baseline and 1 year. RESULTS: The prevalence of hyperhomocysteinemia (tHcy >15 mumol/L) at baseline was 56% in study A and 85% in study B. Baseline tHcy was negatively correlated with measures of body fat and dietary protein intake. Folate, vitamin B(12), and GFR were the major determinants of tHcy levels. Of the patients with hyperhomocysteinemia at baseline, 49% and 24% reduced their tHcy levels at 1 year to < or =15 micromol/L in study A and study B, respectively. There was no association between dietary protein intake and odds of developing hyperhomocysteinemia at 1 year in study A (P= 0.94) or study B (P= 0.10). CONCLUSION: Hyperhomocysteinemia is partly amenable to correction by vitamin supplementation in CKD stages 3 and 4. There is insufficient evidence to suggest that low tHcy is associated with poor nutritional status in the MDRD Study cohort. B vitamins and GFR, but not dietary protein, are the major determinants of tHcy in this patient population. PMID- 15780110 TI - Lipid abnormalities associated with urinary albumin excretion rate in Taiwanese type 2 diabetic patients. AB - BACKGROUND: The purpose of this study was to examine the lipid abnormalities associated with urinary albumin excretion rate (UAER) in type 2 diabetic patients. METHODS: A total of 275 (122 men and 153 women; aged 60.6 +/- 11.1 years) patients were selected with stringent criteria to prevent confounders. Normoalbuminuria (N= 152) and albuminuria (N= 123) were defined as urinary albumin-to-creatinine ratio (ACR) of <30 and > or =30 microg/mg, respectively. Total cholesterol, triglycerides, low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol, and apolipoproteins A1 (ApoA1) and B (ApoB) were measured and non-HDL cholesterol calculated. The subjects were divided into four phenotypes based on triglycerides (<1.5 or > or =1.5 mmol/L) and ApoB (<1.2 or > or =1.2 g/L). RESULTS: Total cholesterol, ApoB, and non-HDL cholesterol were significantly (P < 0.05) higher in patients with albuminuria. For quartiles of the lipid parameters, prevalences of albuminuria showed significant association with ApoB and non-HDL cholesterol (P trend <0.05). After adjusting for age, systolic blood pressure and hemoglobin A(1c) (HbA(1c)) correlation coefficients between the natural logarithm (ln) ACR and lipid parameters, odds ratios for albuminuria, and standardized regression coefficients for ln ACR, were significant for total cholesterol, ApoB and non-HDL cholesterol in all subjects and in men, but only ApoB was significant in women. For patients with normoalbuminuria, frequencies of normotriglycerides/normo-ApoB, hypertriglycerides/normo-ApoB, normotriglycerides/hyper-ApoB, and hypertriglycerides/hyper-ApoB were 44.7%, 28.9%, 10.5%, and 15.8%, respectively; and were 30.1%, 19.5%, 15.4%, and 35.0% for patients with albuminuria (P < 0.001). The respective adjusted odds ratio for albuminuria for the four phenotypes was 1.00, 1.04 (0.54 to 2.00), 2.25 (1.02 to 5.00), and 3.38 (1.75 to 6.53). CONCLUSION: Increased UAER is associated with ApoB-containing lipoproteins and the phenotype of hypertriglycerides/hyper-ApoB is associated with the highest risk of albuminuria. The surrogate marker of non-HDL cholesterol for ApoB is more applicable to the diabetic men. PMID- 15780111 TI - Regular ultrasonographic screening significantly prolongs patency of PTFE grafts. AB - BACKGROUND: Polytetrafluoroethylene (PTFE) dialysis grafts have considerably shorter patency than native arteriovenous fistulas, despite the use of a complex of screening monitoring methods (venous pressure, access flow). PTFE grafts are used often in subjects with depleted subcutaneous veins after previous abandoned accesses, so keeping the access patent is crucial. We hypothesized that regular duplex Doppler ultrasound screening for access stenoses, together with their sooner treatment, would prolong PTFE graft patency. METHODS: We performed a randomized, prospective study of PTFE grafts' cumulative patency in 192 subjects. In group 1, regular ultrasound examinations performed every 3 months was added to traditional screening (i.e., regular access examination at hemodialysis unit, monitoring of venous pressure and access flow). Group 2 was screened only traditionally (without ultrasound). Interventions of suspected stenoses were indicated by nephrologists, vascular surgeon, and, in group 1, also by ultrasonography. Classic ultrasound criteria for significant stenosis were used, even if the access flow had not been decreased. The mean follow-up lasted 392 +/- 430 days. RESULTS: Groups were similar with respect to age, gender, diabetes status, and number of previous abandoned accesses. Group 1 had significantly longer access patency (P < 0.001). Number of interventions per graft was 2.1 +/- 1.8 and 1.3 +/- 1.0 in group 1 and group 2. CONCLUSIONS: Regular screening duplex Doppler ultrasonography results in significantly longer PTFE graft patency due to early detection of access stenosis and, thus, more frequent elective interventions of access stenoses. PMID- 15780112 TI - Benefits of switching from a conventional to a low-GDP bicarbonate/lactate buffered dialysis solution in a rat model. AB - BACKGROUND: Long-term exposure to standard peritoneal dialysis fluid (PDF) results in alterations in peritoneal morphology and function. Studies investigating the long-term effects on the peritoneum of a low-glucose degradation product (GDP) bicarbonate/lactate-buffered PDF demonstrated its superior biocompatibility. We examined the potential of the low-GDP bicarbonate/lactate-buffered solution to reverse or reduce standard PDF-induced peritoneal alterations. METHODS: Female Wistar rats received twice daily intraperitoneal infusions with either a lactate-buffered solution with 3.86% glucose at pH 5.5 (Dianeal, referred to as standard PDF), or a low-GDP bicarbonate/lactate-buffered solution with 3.86% glucose at physiologic pH (Physioneal, referred to as bicarbonate/lactate PDF) for different periods of time: (1) 12 weeks Dianeal (N= 9); (2) 12 weeks Physioneal (N= 9); (3) 20 weeks Dianeal (N= 11); (4) 20 weeks Physioneal (N= 10); (5) 12 weeks Dianeal followed by 8 weeks Physioneal (N= 10). RESULTS: Chronic standard PDF exposure resulted in loss of ultrafiltration capacity, increased VEGF expression and vascular density, higher advanced glycation end product (AGE) accumulation, up-regulation of TGF beta expression, and development of fibrosis compared to low-GDP bicarbonate/lactate-buffered PDF. The PDF-induced alterations were time dependent. Crossover from standard PDF to low-GDP bicarbonate/lactate PDF resulted in a less impaired ultrafiltration (UF), less pronounced VEGF expression and neoangiogenesis, and less severe AGE accumulation, TGF-beta expression, and fibrosis compared to continuous standard PDF exposure for 20 weeks. CONCLUSION: Low-GDP bicarbonate/lactate-buffered PDF has the potential to slow down standard PDF-induced peritoneal membrane damage. PMID- 15780113 TI - Kinetic behavior of urea is different from that of other water-soluble compounds: the case of the guanidino compounds. AB - BACKGROUND: Although patients with renal failure retain a large variety of solutes, urea is virtually the only currently applied marker for adequacy of dialysis. Only a limited number of other compounds have up until now been investigated regarding their intradialytic kinetics. Scant data suggest that large solutes show a kinetic behavior that is different from urea. The question investigated in this study was whether other small water-soluble solutes, such as some guanidino compounds, show a kinetic behavior comparable or dissimilar to that of urea. METHODS: This study included 7 stable conventional hemodialysis patients without native kidney function undergoing low flux polysulphone dialysis (F8 and F10HPS). Blood samples were collected from the inlet and outlet bloodlines immediately before the dialysis session, after 5, 15, 30, 120 minutes, and immediately after discontinuation of the session. Plasma concentrations of urea, creatinine (CTN), creatine (CT), guanidinosuccinic acid (GSA), guanidinoacetic acid (GAA), guanidine (G), and methylguanidine (MG) were used to calculate corresponding dialyzer clearances. A two-pool kinetic model was fitted to the measured plasma concentration profiles, resulting in the calculation of the perfused volume (V(1)), the total distribution volume (V(tot)), and the intercompartmental clearance (K(12)); solute generation and overall ultrafiltration were determined independently. RESULTS: No significant differences were observed between V(1) and K(12) for urea (6.4 +/- 3.3 L and 822 +/- 345 mL/min, respectively) and for the guanidino compounds. However, with respect to V(tot), GSA was distributed in a smaller volume (30.6 +/- 4.2 L) compared to urea (42.7 +/- 6.0L) (P < 0.001), while CTN, CT, GAA, G, and MG showed significantly higher volumes (54.0 +/- 5.9 L, 98.0 +/- 52.3 L, 123.8 +/- 66.9 L, 89.7 +/- 21.4 L, 102.6 +/- 33.9 L, respectively; P= 0.004, = 0.033, = 0.003, < 0.001, = 0.001, respectively). These differences resulted in divergent effective solute removal: 67% (urea), 58% (CTN), 42% (CT), 76% (GSA), 37% (GAA), 43% (G), and 42% (MG). CONCLUSION: The kinetics of the guanidino compounds under study are different from that of urea; hence, urea kinetics are not representative for the removal of other uremic solutes, even if they are small and water-soluble like urea. PMID- 15780114 TI - Coronary calcification in hemodialysis patients: the contribution of traditional and uremia-related risk factors. AB - BACKGROUND: Coronary artery calcification is a common feature of atherosclerosis, occurring in 90% of angiographically significant lesions. There is recent evidence that coronary artery calcification is frequent in hemodialysis patients and it has been suggested that this increased incidence may be associated to uremia-related factors. The development and progression of coronary artery calcification is similar to osteogenesis. The aim of this study was to evaluate the relationship between coronary artery calcification, uremia-related factors, and bone histomorphometry in hemodialysis patients. METHODS: A total of 101 hemodialysis patients were assessed for biochemical markers of inflammation, oxidative stress, and bone metabolism. Subsequently, they were submitted to multislice coronary tomography (MSCT) and transiliac bone biopsy. RESULTS: The median calcium score was 116.2 (range 0 to 5547). Fifty-two percent of the patients showed moderate and severe coronary artery calcification, 20% had calcium scores greater than 1000. In univariate analysis, age (r= 0.57, P < 0.000001), osteoprotegerin (OPG) (r= 0.44, P= 0.00002), and body mass index (BMI) (r= 0.24, P= 0.01) correlated positively with calcium score. Bone trabecular volume and trabecular thickness correlated negatively with calcium score (r= 0.24, P= 0.02; r=-0.22, P= 0.03). There was a correlation of borderline significance between calcium score and C-reactive protein (CRP) (r= 0.18, P= 0.062). The multiple linear regression analysis identified OPG as the only variable independently associated with coronary artery calcification. CONCLUSION: Coronary artery calcification is highly prevalent in the hemodialysis population and is associated with older age, higher BMI, inflammation and reduced trabecular bone volume. Higher OPG is independently associated with coronary artery calcification and may represent an incomplete self-defensive response to the progression of atherosclerosis in hemodialysis patients. PMID- 15780115 TI - Lectin-like oxidized low-density lipoprotein (LDL) receptor (LOX-1)-mediated pathway and vascular oxidative injury in older-age rat renal transplants. AB - BACKGROUND: Older-age renal allografts are associated with inferior survival; however, the mechanisms are unclear. Reactive oxygen species participate in aging and in chronic vascular disease. We investigated how mediators of oxidative stress may increase allograft susceptibility to vascular injury. METHODS: We employed the low-responder allogeneic F344-to-Lew rat renal transplantation model. We used nonimmunosuppressed young (donors and recipients aged 12 weeks), old (donors and recipients aged 52 weeks), and old-to-young animal (donors aged 52 weeks and recipients aged 12 weeks) combinations. Grafts were transplanted after 2 hours cold preservation in University of Wisconsin solution and harvested 1, 2, 7 and 10 days later. Additionally, old animals receiving continuous 1.5 mg/kg cyclosporine (CyA) immunosuppression were included. Renal allograft pathology was scored according to Banff criteria. We studied intragraft vascular adhesion molecule-1 (VCAM-1), lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (LOX-1), and hypochlorite-modified LDL expression as well as ED-1+ monocytes/macrophages and CD8+ lymphocyte infiltration. Intragraft in situ superoxide anion radical production was determined with dihydroethidium assay on cryosections. RESULTS: During the first 2 posttransplant days, old transplants demonstrated higher functional impairment and increased oxidative stress, while young transplant had higher ED-1+ monocytes/macrophage infiltration and VCAM-1 expression. The degree of VCAM-1 expression and ED-1+ monocytes/macrophage and CD8+ lymphocyte infiltration correlated at later time points directly with the transplant age. VCAM-1 and LOX-1 staining were localized predominantly on the endothelium of arterial vessels, shifting the distribution to vascular smooth muscle layer strongly dependent on donor age and the grade of vascular injury. LOX-1 staining colocalized with hypochlorite-modified epitopes in the media of injured arteries. We measured increased in situ superoxide anion radical production in corresponding areas. Immunosuppression with CyA had no protective effect on vascular injury and LOX-1 expression. CONCLUSION: Induction of LOX-1 related oxidation pathways and increased susceptibility to oxidative stress could play an important role in promoting vascular injury in old renal transplants independent of the recipient age. PMID- 15780116 TI - Evaluation of pre-implantation kidney biopsies: comparison of Banff criteria to a morphometric approach. AB - BACKGROUND: Donor glomerulosclerosis, interstitial fibrosis, and fibrous intimal thickening correlate with graft outcome. We evaluate chronic lesions in donor biopsies according to Banff criteria and with a morphometric technique to ascertain their predictive value on graft outcome. METHODS: We evaluated 77 cadaveric donor biopsies according to Banff criteria. Glomerulosclerosis was expressed as the percentage of global sclerotic glomeruli. The following morphometric parameters were obtained: cortical interstitial volume fraction (Vvint/c), cortical glomerular volume fraction (Vvglom/c), mean glomerular volume (Vg), mean and maximal intimal arterial volume fraction (Vvintima/art), and Vvintima/art of the largest artery. We evaluated the correlation of histologic lesions with delayed graft function, 3 months' glomerular filtration rate (GFR), and death-censored graft survival. RESULTS: Multivariate logistic regression showed that delayed graft function was associated with cv score [relative risk (RR) 4.2 and 95% CI 1.1 to 16.0) and glomerulosclerosis (RR 1.06 and 95% CI 1.01 to 1.13). Stepwise regression showed that Vvint/c and glomerulosclerosis were independent predictors of 3 months' GFR (R= 0.62, P= 0.0001). Repeated analysis not considering morphometric parameters showed that glomerulosclerosis, cv score and ci score were independent predictors of 3 months' GFR (R= 0.64, P= 0.0001). A donor chronic damage score was generated considering glomerulosclerosis, cv score and ci score. This score after adjusting for clinical variables was associated with 3 months' GFR (R= 0.71, P < 0.0001) and death-censored graft survival (RR 2.2 and 95% CI 1.3 to 3.7). CONCLUSION: Combined evaluation of donor glomerulosclerosis, chronic vascular and interstitial damage according to Banff criteria allows a precise prediction of graft outcome. Morphometric evaluation of donor biopsies does not improve the predictive value of semiquantitative grading. PMID- 15780117 TI - Predilution hemodiafiltration displays no hemodynamic advantage over low-flux hemodialysis under matched conditions. AB - BACKGROUND: It is the prevailing view that convective dialysis techniques stabilize blood pressure. The aim of this study was to compare the intrasession hemodynamics during high-dose predilution hemodiafiltration (HDF) and low-flux hemodialsis, under strict controlled conditions. METHODS: Twelve stable hemodialysis patients were investigated in a randomized crossover blinded controlled trial. The patients were allocated to one session of predilution HDF (substitution fluid 1.20 +/- 0.10 L/kg body weight) and one session of hemodialysis at 4(1/2) hours. To eliminate confounding factors, dialysis dose, ultrafiltration volume and arterial temperature were matched. At the start of the dialysis the patients' core temperature was "locked" by an automatic feedback system regulating the dialysate temperature; thereby, patients' temperature was kept stable throughout the whole treatment. The calcium-ion concentration in the substitution/dialysis fluid was 1.25 mmol/L. Cardiac output was measured hourly by the ultrasound velocity dilution method. RESULTS: Mean blood pressure, cardiac output, stroke volume, cardiac work, and relative blood volume was significantly reduced in both treatments. Total peripheral resistance increased significantly in both groups. Ultrafiltration volume, cardiopulmonary recirculation, Kt/V, and total energy transfer were similar for hemodialysis and HDF. The pulse rate showed no significant change throughout both sessions. No significant differences were revealed between hemodialysis and HDF. CONCLUSION: The hemodynamics of predilution HDF and low-flux hemodialysis displayed a similar profile during matched conditions. An acute circulatory benefit of convective solute removal over diffusive could not be demonstrated. PMID- 15780118 TI - Longitudinal membrane function in functionally anuric patients treated with APD: data from EAPOS on the effects of glucose and icodextrin prescription. AB - BACKGROUND: Peritoneal dialysis is associated with changes in membrane function that can lead eventually to ultrafiltration (UF) failure. Factors driving these changes are thought to include hypertonic glucose exposure, but previously reported associations are confounded by the presence of residual renal function. METHODS: Longitudinal membrane function (solute transport and UF capacity) were measured annually in a prospective cohort of 177 functionally anuric patients as part of the European Automated Peritoneal Dialysis Outcomes Study (EAPOS). Subgroup analysis was performed according to glucose exposure and icodextrin use at baseline. RESULTS: The whole cohort experienced an increase in solute transport and reduction in UF capacity at 12 and 24 months that could not be explained by informative censoring. These changes were accelerated and more severe in patients using either 2.27% or 3.86% glucose, or those not using icodextrin at baseline. These differences could not be explained by age, comorbidity score, previous time spent on renal replacement, differential dropout from the study, peritonitis rates, or, by definition, residual renal function. Patients using icodextrin at baseline had worse membrane function and were more likely to be diabetic. There was an association between membrane function changes and achieved 24-hour ultrafiltration over the 2-year study period. CONCLUSION: Anuric automated peritoneal dialysis (APD) patients experience significant detrimental changes in membrane function over a relatively short time period. Glucose appears to enhance these changes independent of residual renal function. Icodextrin use in these circumstances is associated with less deterioration in membrane function. PMID- 15780119 TI - BCA-1/CXCL13 expression is associated with CXCR5-positive B-cell cluster formation in acute renal transplant rejection. AB - BACKGROUND: Recent studies showed a crucial role for B cells in acute renal allograft rejection. It remains largely unknown, however, which mechanisms lead to the B-cell recruitment into the allograft. The chemokine CXCL13 and its corresponding receptor CXCR5 play a central role in B-cell trafficking to secondary lymphatic tissue and ectopic B-cell clusters in rheumatoid arthritis. We therefore investigated the potential role of CXCL13 and CXCR5 in formation of B-cell clusters in renal transplant rejection. METHODS: Serial immunohistochemical staining for CXCL13, CXCR5, and CD20 was carried out in protocol biopsies of 23 patients obtained between day 4 and day 9 after renal transplantation. Intragraft mRNA expression of CXCL13 was assessed by real-time polymerase chain reaction (PCR) analysis. RESULTS: Of 23 kidney biopsies obtained between days 4 and 9 after renal transplantation, 13 revealed an acute rejection. Four of these patients showed a substantial infiltration of the transplant with cluster-forming B cells. By immunohistochemistry CXCL13 and the corresponding receptor CXCR5 were exclusively detected in areas of B-cell clusters. Intrarenal CXCL13 mRNA expression was 27-fold higher in transplants with B-cell clusters compared to rejecting allografts without B-cell accumulation (P= 0.011). CONCLUSION: We describe a striking colocalization of CXCL13 expression with CXCR5 and CD20-positive B cells in renal transplants undergoing rejection. This is the first study demonstrating a potential role of CXCL13 and its specific receptor CXCR5 in recruitment of B cells in renal allograft rejection. PMID- 15780120 TI - Safety and success of kidney transplantation and concomitant immunosuppression in HIV-positive patients. AB - BACKGROUND: Human immunodeficiency virus-associated nephropathy (HIVAN) has become the third leading cause of end-stage renal disease (ESRD) in African Americans, and is expected to grow exponentially. Highly active antiretroviral therapy (HAART) has significantly prolonged the survival of patients with HIV infection. Despite the growing number of HIV-positive dialysis patients with prolonged life expectancy, kidney transplantation with immunosuppression has been declined because it is considered a waste of scarce donor kidneys due to potential increases in morbidity and mortality. METHODS: The institutional review board of Drexel University College of Medicine and Hahnemann University Hospital approved this prospective study. The aim was to find out safety and success of kidney transplantation, and the effect of immunosuppression on HIV infection. Forty HIV-positive dialysis patients received kidney transplantation between February 2001 and January 2004. Patient inclusion criteria were maintenance of HAART, plasma HIV-1 RNA of <400 copies/mL, absolute CD4 counts of 200 cells/muL or more. Immunosuppression was basiliximab induction and maintenance with cyclosporine, sirolimus, and steroids. HAART was continued post-transplant. Acute rejections were diagnosed by biopsy and treated with methylprednisolone. Surveillance biopsies were completed at 1, 6, 12, and 24 months, and evaluated for subclinical acute rejection, chronic allograft nephropathy, and HIVAN. RESULTS: One- and 2-year actuarial patient survival was 85% and 82%, respectively, and graft survival was 75% and 71%, respectively. Plasma HIV-1 RNA remained undetectable, and CD4 counts remained in excess of 400 cells per muL with no evidence of AIDS for up to 2 years. CONCLUSION: One- and 2-year graft survival is comparable to other high-risk populations receiving kidney transplantation. One- and 2-year patient survival is higher than HIV patients maintained on dialysis. Immunosuppression does not adversely affect HIV recipients maintained on HAART in the short term. PMID- 15780121 TI - Nature and nurture on phenotypic variability of autosomal dominant polycystic kidney disease. PMID- 15780122 TI - Nephropathy in the context of HIV infection. PMID- 15780124 TI - Effectiveness and safety of dialysis vascular access procedures performed by interventional nephrologists. PMID- 15780126 TI - Interpretation of erythropoietin levels in patients with various degrees of renal anemia. PMID- 15780127 TI - Economic impact of sevelamer in patients with ESRD. PMID- 15780128 TI - Opportunity and cost of sevelamer in dialysis patients. PMID- 15780129 TI - Body mass index and survival differences in dialysis patients. PMID- 15780130 TI - Autosomal-recessive polycystic kidney disease. PMID- 15780132 TI - A topological model of biofeedback based on catecholamine interactions. AB - BACKGROUND: The present paper describes a topological model of biofeedback. This model incorporates input from a sensory organ and a transduction phase mediated through catecholamine production in the feedback path. The transduction phase comprises both conservative and dissipative systems, from which the appropriate output is combined in a closed loop. RESULTS: The model has been simulated in MATLAB 6.0 R12 in order to facilitate a comprehensive understanding of the complex biofeedback phenomena concomitant with the transduction phases associated with migraine and with psychosomatic diseases involving digestive disorders. CONCLUSION: The complexity of the biological system influences the transduction phase and nature of the system response, which is consequent on the activation of smooth muscles by sympathetic and parasympathetic stimulation. PMID- 15780133 TI - Surgical revascularization versus amputation for peripheral vascular disease in dialysis patients: a cohort study. AB - BACKGROUND: Surgical treatment of peripheral vascular disease (PVD) in dialysis patients is controversial. METHODS: We examined the post-operative morbidity and mortality of surgical revascularization or amputation for PVD in a retrospective analysis of United States Renal Data System. Propensity scores for undergoing amputation were derived from a multivariable logistic regression model of amputation. RESULTS: Of the Medicare patients initiated on dialysis from Jan 1, 1995 to Dec 31, 1999, patients underwent surgical revascularization (n = 1,896) or amputation (n = 2,046) in the first 6 months following initiation of dialysis were studied. In the logistic regression model, compared to claudication, presence of gangrene had a strong association with amputation [odds ratio (OR) 19.0, 95% CI (confidence interval) 13.86-25.95]. The odds of dying within 30 days and within 1 year were higher (30 day OR: 1.85, 95% CI: 1.45-2.36; 1 yr OR: 1.46, 95% CI: 1.25-1.71) in the amputation group in logistic regression model adjusted for propensity scores and other baseline factors. Amputation was associated with increased odds of death in patients with low likelihood of amputation (< 33rd percentile of propensity score) and moderate likelihood of amputation (33rd to 66th percentile) but not in high likelihood group (> 66th percentile). The number of hospital days in the amputation and revascularization groups was not different. CONCLUSION: Amputation might be associated with higher mortality in dialysis patients. Where feasible, revascularization might be preferable over amputation in dialysis patients. PMID- 15780134 TI - A standard curve based method for relative real time PCR data processing. AB - BACKGROUND: Currently real time PCR is the most precise method by which to measure gene expression. The method generates a large amount of raw numerical data and processing may notably influence final results. The data processing is based either on standard curves or on PCR efficiency assessment. At the moment, the PCR efficiency approach is preferred in relative PCR whilst the standard curve is often used for absolute PCR. However, there are no barriers to employ standard curves for relative PCR. This article provides an implementation of the standard curve method and discusses its advantages and limitations in relative real time PCR. RESULTS: We designed a procedure for data processing in relative real time PCR. The procedure completely avoids PCR efficiency assessment, minimizes operator involvement and provides a statistical assessment of intra assay variation. The procedure includes the following steps. (I) Noise is filtered from raw fluorescence readings by smoothing, baseline subtraction and amplitude normalization. (II) The optimal threshold is selected automatically from regression parameters of the standard curve. (III) Crossing points (CPs) are derived directly from coordinates of points where the threshold line crosses fluorescence plots obtained after the noise filtering. (IV) The means and their variances are calculated for CPs in PCR replicas. (V) The final results are derived from the CPs' means. The CPs' variances are traced to results by the law of error propagation. A detailed description and analysis of this data processing is provided. The limitations associated with the use of parametric statistical methods and amplitude normalization are specifically analyzed and found fit to the routine laboratory practice. Different options are discussed for aggregation of data obtained from multiple reference genes. CONCLUSION: A standard curve based procedure for PCR data processing has been compiled and validated. It illustrates that standard curve design remains a reliable and simple alternative to the PCR-efficiency based calculations in relative real time PCR. PMID- 15780136 TI - Worldwide trends in quantity and quality of published articles in the field of infectious diseases. AB - BACKGROUND: Trying to confront with the widespread burden of infectious diseases, the society worldwide invests considerably on research. We evaluated the contribution of different world regions in research production in Infectious Diseases. METHODS: Using the online Pubmed database we retrieved articles from 38 journals included in the "Infectious Diseases" category of the "Journal Citation Reports" database of the Institute for Scientific Information for the period 1995 2002. The world was divided into 9 regions based on geographic, economic and scientific criteria. Using an elaborate retrieval system we obtained data on published articles from different world regions. In our evaluation we introduced an estimate of both quantity and quality of research produced from each world region per year using: (1) the total number of publications, (2) the mean impact factor of publications, and (3) the product of the above two parameters. RESULTS: Data on the country of origin of the research was available for 45,232 out of 45,922 retrieved articles (98.5 %). USA and Western Europe are by far the most productive regions concerning publications of research articles. However, the rate of increase in the production of articles was higher in Eastern Europe, Africa, Latin America and the Caribbean, and Asia during the study period. The mean impact factor is highest for articles originating in the USA (3.42), while it was 2.82 for Western Europe and 2.73 for the rest of the world (7 regions combined). CONCLUSION: USA and Western Europe make up a striking 80% of the world's research production in Infectious Diseases in terms of both quantity and quality. However, all world regions achieved a gradual increase in the production of Infectious Diseases articles, with the regions ranking lower at present displaying the highest rate of increase. PMID- 15780135 TI - A genetic study of autism in Costa Rica: multiple variables affecting IQ scores observed in a preliminary sample of autistic cases. AB - BACKGROUND: Autism is a heritable developmental disorder of communication and socialization that has not been well studied in Hispanic populations. Therefore, we are collecting and evaluating all possible cases of autism from a population isolate in the Central Valley of Costa Rica (CVCR) for a clinical and genetic study. METHODS: We are assessing all subjects and parents, as appropriate, using the newly translated Spanish versions of the Autism Diagnostic Interview-Revised (ADI-R) and the Autism Diagnostic Observation Schedule (ADOS) as well as tests of intelligence and adaptive behavior. Detailed obstetric and family medical/psychiatric histories are taken. All cases are tested for Fragile X and will be extensively evaluated for cytogenetic abnormalities. RESULTS: To date we have obtained clinical evaluations on over 76 cases of possible autism referred to our study and report data for the initial 35 complete cases. The mean age of the probands is 6.7 years, and 31 of the 35 cases are male. Twenty-one of the cases have IQs <50 and only 6 cases have IQs > or = 70. Over half of the mothers had complications during pregnancy and/or delivery. No cases have tested positively for Fragile X or PKU. Chromosomal G-banding is not yet complete for all cases. CONCLUSION: Diagnostic data gathered on cases of autism in the CVCR using Spanish versions of the ADI-R and ADOS look similar to that generated by studies of English-speaking cases. However, only 17% of our cases have IQs within the normal range, compared to the figure of 25% seen in most studies. This result reflects an ascertainment bias in that only severe cases of autism come to treatment in the CVCR because there are no government-sponsored support programs or early intervention programs providing an incentive to diagnose autism. The severity of mental retardation seen in most of our cases may also be exaggerated by the lack of early intervention programs and the use of IQ tests without Costa Rican norms. Still, we must formally train healthcare providers and teachers to recognize and refer autistic cases with normal or near normal IQs that are not seen in treatment. PMID- 15780137 TI - Evolutionary models for insertions and deletions in a probabilistic modeling framework. AB - BACKGROUND: Probabilistic models for sequence comparison (such as hidden Markov models and pair hidden Markov models for proteins and mRNAs, or their context free grammar counterparts for structural RNAs) often assume a fixed degree of divergence. Ideally we would like these models to be conditional on evolutionary divergence time. Probabilistic models of substitution events are well established, but there has not been a completely satisfactory theoretical framework for modeling insertion and deletion events. RESULTS: I have developed a method for extending standard Markov substitution models to include gap characters, and another method for the evolution of state transition probabilities in a probabilistic model. These methods use instantaneous rate matrices in a way that is more general than those used for substitution processes, and are sufficient to provide time-dependent models for standard linear and affine gap penalties, respectively. Given a probabilistic model, we can make all of its emission probabilities (including gap characters) and all its transition probabilities conditional on a chosen divergence time. To do this, we only need to know the parameters of the model at one particular divergence time instance, as well as the parameters of the model at the two extremes of zero and infinite divergence. I have implemented these methods in a new generation of the RNA genefinder QRNA (eQRNA). CONCLUSION: These methods can be applied to incorporate evolutionary models of insertions and deletions into any hidden Markov model or stochastic context-free grammar, in a pair or profile form, for sequence modeling. PMID- 15780139 TI - Evaluating an intervention to reduce fear of falling and associated activity restriction in elderly persons: design of a randomised controlled trial [ISRCTN43792817]. AB - BACKGROUND: Fear of falling and associated activity restriction is common in older persons living in the community. Adverse consequences of fear of falling and associated activity restriction, like functional decline and falls, may have a major impact on physical, mental and social functioning of these persons. This paper presents the design of a trial evaluating a cognitive behavioural group intervention to reduce fear of falling and associated activity restriction in older persons living in the community. METHODS/DESIGN: A two-group randomised controlled trial was developed to evaluate the intervention. Persons 70 years of age or over and still living in the community were eligible for study if they experienced at least some fear of falling and associated activity restriction. A random community sample of elderly people was screened for eligibility; those eligible for study were measured at baseline and were subsequently allocated to the intervention or control group. Follow-up measurements were carried out directly after the intervention period, and then at six months and 12 months after the intervention. People allocated to the intervention group were invited to participate in eight weekly sessions of two hours each and a booster session. This booster session was conducted before the follow-up measurement at six months after the intervention. People allocated to the control group received no intervention as a result of this trial. Both an effect evaluation and a process evaluation were performed. The primary outcome measures of the effect evaluation are fear of falling, avoidance of activity due to fear of falling, and daily activity. The secondary outcome measures are perceived general health, self-rated life satisfaction, activities of daily life, feelings of anxiety, symptoms of depression, social support interactions, feelings of loneliness, falls, perceived consequences of falling, and perceived risk of falling. The outcomes of the process evaluation comprise the performance of the intervention according to protocol, the attendance and adherence of participants, and the participants' and facilitators' opinion about the intervention. Data of the effect evaluation will be analysed according the intention-to-treat and on-treatment principle. Data of the process evaluation will be analysed using descriptive techniques. PMID- 15780141 TI - Therapeutic targets for HIV-1 infection in the host proteome. AB - BACKGROUND: Despite the success of HAART, patients often stop treatment due to the inception of side effects. Furthermore, viral resistance often develops, making one or more of the drugs ineffective. Identification of novel targets for therapy that may not develop resistance is sorely needed. Therefore, to identify cellular proteins that may be up-regulated in HIV infection and play a role in infection, we analyzed the effects of Tat on cellular gene expression during various phases of the cell cycle. RESULTS: SOM and k-means clustering analyses revealed a dramatic alteration in transcriptional activity at the G1/S checkpoint. Tat regulates the expression of a variety of gene ontologies, including DNA-binding proteins, receptors, and membrane proteins. Using siRNA to knock down expression of several gene targets, we show that an Oct1/2 binding protein, an HIV Rev binding protein, cyclin A, and PPGB, a cathepsin that binds NA, are important for viral replication following induction from latency and de novo infection of PBMCs. CONCLUSION: Based on exhaustive and stringent data analysis, we have compiled a list of gene products that may serve as potential therapeutic targets for the inhibition of HIV-1 replication. Several genes have been established as important for HIV-1 infection and replication, including Pou2AF1 (OBF-1), complement factor H related 3, CD4 receptor, ICAM-1, NA, and cyclin A1. There were also several genes whose role in relation to HIV-1 infection have not been established and may also be novel and efficacious therapeutic targets and thus necessitate further study. Importantly, targeting certain cellular protein kinases, receptors, membrane proteins, and/or cytokines/chemokines may result in adverse effects. If there is the presence of two or more proteins with similar functions, where only one protein is critical for HIV-1 transcription, and thus, targeted, we may decrease the chance of developing treatments with negative side effects. PMID- 15780143 TI - Values for preventing influenza-related morbidity and vaccine adverse events in children. AB - BACKGROUND: Influenza vaccination recently has been recommended for children 6-23 months old, but is not currently recommended for routine use in non-high-risk older children. Information on disease impact, costs, benefits, risks, and community preferences could help guide decisions about which age and risk groups should be vaccinated and strategies for improving coverage. The objective of this study was to measure preferences and willingness-to-pay for changes in health related quality of life associated with uncomplicated influenza and two rarely occurring vaccination-related adverse events (anaphylaxis and Guillain-Barre syndrome) in children. METHODS: We conducted telephone interviews with adult members selected at random from a large New England HMO (n = 112). Respondents were given descriptions of four health outcomes: uncomplicated influenza in a hypothetical 1-year-old child of their own, uncomplicated influenza in a hypothetical 14-year-old child of their own, anaphylaxis following vaccination, and Guillain-Barre syndrome. "Uncomplicated influenza" did not require a physician's visit or hospitalization. Preferences (values) for these health outcomes were measured using time-tradeoff and willingness-to-pay questions. Time tradeoff questions asked the adult to assume they had a child and to consider how much time from the end of their own life they would be willing to surrender to avoid the health outcome in the child. RESULTS: Respondents said they would give a median of zero days of their lives to prevent an episode of uncomplicated influenza in either their (hypothetical) 1-year-old or 14-year-old, 30 days to prevent an episode of vaccination-related anaphylaxis, and 3 years to prevent a vaccination-related case of Guillain-Barre syndrome. Median willingness-to-pay to prevent uncomplicated influenza in a 1-year-old was $175, uncomplicated influenza in a 14-year-old was $100, anaphylaxis $400, and Guillain-Barre syndrome $4000. The median willingness-to-pay for an influenza vaccination for their children with no risk of anaphylaxis or Guillain-Barre syndrome was $50 and $100, respectively. CONCLUSION: Most respondents said they would not be willing to trade any time from their own lives to prevent uncomplicated influenza in a child of their own, and the time traded did not vary by the age of the hypothetical affected child. However, adults did indicate a willingness-to-pay to prevent uncomplicated influenza in children, and that they would give more money to prevent the illness in a 1-year-old than in a 14-year-old. Respondents also indicated a willingness to pay a premium for a vaccine without any risk of severe complications. PMID- 15780142 TI - A gene expression fingerprint of C. elegans embryonic motor neurons. AB - BACKGROUND: Differential gene expression specifies the highly diverse cell types that constitute the nervous system. With its sequenced genome and simple, well defined neuroanatomy, the nematode C. elegans is a useful model system in which to correlate gene expression with neuron identity. The UNC-4 transcription factor is expressed in thirteen embryonic motor neurons where it specifies axonal morphology and synaptic function. These cells can be marked with an unc-4::GFP reporter transgene. Here we describe a powerful strategy, Micro-Array Profiling of C. elegans cells (MAPCeL), and confirm that this approach provides a comprehensive gene expression profile of unc-4::GFP motor neurons in vivo. RESULTS: Fluorescence Activated Cell Sorting (FACS) was used to isolate unc 4::GFP neurons from primary cultures of C. elegans embryonic cells. Microarray experiments detected 6,217 unique transcripts of which approximately 1,000 are enriched in unc-4::GFP neurons relative to the average nematode embryonic cell. The reliability of these data was validated by the detection of known cell specific transcripts and by expression in UNC-4 motor neurons of GFP reporters derived from the enriched data set. In addition to genes involved in neurotransmitter packaging and release, the microarray data include transcripts for receptors to a remarkably wide variety of signaling molecules. The added presence of a robust array of G-protein pathway components is indicative of complex and highly integrated mechanisms for modulating motor neuron activity. Over half of the enriched genes (537) have human homologs, a finding that could reflect substantial overlap with the gene expression repertoire of mammalian motor neurons. CONCLUSION: We have described a microarray-based method, MAPCeL, for profiling gene expression in specific C. elegans motor neurons and provide evidence that this approach can reveal candidate genes for key roles in the differentiation and function of these cells. These methods can now be applied to generate a gene expression map of the C. elegans nervous system. PMID- 15780144 TI - Human immunodeficiency virus type 1 (HIV-1) proviral DNA load in purified CD4+ cells by LightCycler real-time PCR. AB - BACKGROUND: The human immunodeficiency virus type 1 (HIV-1) proviral DNA persists in infected cells, even after prolonged successful HAART. In the present study, a relative quantification assay of HIV-1 proviral DNA by LightCycler real-time PCR based on SYBR Green I detection was developed in comparison to the number of purified CD4+ cells as estimated by the quantification of the beta-globin gene. METHODS: The ability of the designed gag primers to quantify HIV-1 Group M and the PCR efficiency were assessed on HIV-1 reference isolate subtypes A, B, C and D. The 8E5 cell line containing a single defective copy of HIV-1 proviral DNA was used as a standard for both the HIV-1 target gene and the beta-globin reference gene. The assay was applied on thirty consecutive patient samples received for RNA viral load determinations and on retrospective samples from fifteen patients undergoing 2 years of structured treatment interruption (STI). RESULTS: The lower limit of quantification was 50 HIV-1 DNA proviral copies per CD4+ cell sample. The dynamic range was from 50 to 106 HIV-1 DNA copies per CD4+ cell sample with intra- and inter-assay coefficients of variability ranging from 3.1% to 37.1%. The beta-globin reference gene was quantified down to a limit of 1.5 pg of DNA/microl (approximately 5 cells) with intra- and interassay coefficients of variability ranging from 1.8% to 21%. DNA proviral load varies widely among HIV-1 infected patients. Proviral load and plasma viral load rebound were high in STI patients who took longer to achieve an undetectable plasma viral load under therapy. A statistically significant correlation was observed between DNA proviral load and RNA steady state viral load in STI patients (p-value = 0.012). CONCLUSION: We have developed a fast, sensitive and specific relative quantification assay of HIV-1 proviral DNA in purified CD4+ cells. The assay enables the monitoring of HIV-1 proviral load, which may be useful to monitor therapy efficacy especially in patients with undetectable plasma RNA viral load, and allows the exploration of viral reservoirs. PMID- 15780145 TI - Health-state valuations for pertussis: methods for valuing short-term health states. AB - BACKGROUND: The incidence of reported adolescent and adult pertussis continues to rise in the United States. Acellular pertussis vaccines for adolescents and adults have been developed and may be available soon for use in the U.S. Our objectives were: (1) to describe patient valuations of pertussis disease and vaccination; and (2) to compare valuations for short-term and long-term health states associated with pertussis. METHODS: We conducted telephone surveys with 515 adult patients and parents of adolescent patients with pertussis in Massachusetts to determine valuations of pertussis-related health states for disease and vaccination using time trade-off (TTO) and contingent valuation (CV) techniques. Respondents were randomized to complete either a short-term or long term TTO exercise. Discrimination between health states for each valuation technique was assessed using Tukey's method, and valuations for short-term vs. long-term health states were compared using the Wilcoxon rank-sum test. RESULTS: Three hundred three (59%) and 309 (60%) respondents completed and understood the TTO and CV exercises, respectively. Overall, respondents gave lower valuations (lower TTO and higher CV values) to avoid a given state for adolescent/adult disease compared to vaccine adverse events. Infant complications due to pertussis were considered worse than adolescent/adult disease, regardless of the method of valuation. The short-term TTO resulted in lower mean valuations and larger mean differences between health states than the long-term TTO exercise. CONCLUSION: Pertussis was considered worse than adverse events due to vaccination. Short-term health-state valuation is better able to discriminate among health states, which is useful for cost-utility analysis. PMID- 15780147 TI - Focus on the metabolic consequences of long-term treatment with olanzapine, quetiapine and risperidone: are there differences? PMID- 15780146 TI - MBEToolbox: a MATLAB toolbox for sequence data analysis in molecular biology and evolution. AB - BACKGROUND: MATLAB is a high-performance language for technical computing, integrating computation, visualization, and programming in an easy-to-use environment. It has been widely used in many areas, such as mathematics and computation, algorithm development, data acquisition, modeling, simulation, and scientific and engineering graphics. However, few functions are freely available in MATLAB to perform the sequence data analyses specifically required for molecular biology and evolution. RESULTS: We have developed a MATLAB toolbox, called MBEToolbox, aimed at filling this gap by offering efficient implementations of the most needed functions in molecular biology and evolution. It can be used to manipulate aligned sequences, calculate evolutionary distances, estimate synonymous and nonsynonymous substitution rates, and infer phylogenetic trees. Moreover, it provides an extensible, functional framework for users with more specialized requirements to explore and analyze aligned nucleotide or protein sequences from an evolutionary perspective. The full functions in the toolbox are accessible through the command-line for seasoned MATLAB users. A graphical user interface, that may be especially useful for non-specialist end users, is also provided. CONCLUSION: MBEToolbox is a useful tool that can aid in the exploration, interpretation and visualization of data in molecular biology and evolution. The software is publicly available at http://web.hku.hk/~jamescai/mbetoolbox/ and http://bioinformatics.org/project/?group_id=454 PMID- 15780149 TI - Food security for all. PMID- 15780150 TI - Mealtimes as active processes in long-term care facilities. AB - Mealtimes are central to the nutritional care of residents in long-term care facilities. There has been little Canadian research to guide interdisciplinary practice around mealtimes. This study included a grounded theory approach to explore mealtime experiences of 20 people with dementia living in two long-term care facilities, and the meal-related care they received from registered nurses, health care aides, and dietitians. Theoretical sampling directed the collection and analysis of data from mealtime observations in special care units and key informant interviews with care providers. The constant comparison method was used to analyze and conceptualize the data. A substantive theory emerged with three key themes: 1. Each mealtime is a unique process embedded within a long-term care facility's environment. 2. Residents are central to the process through their actions (i.e., arriving, eating, waiting, socializing, leaving, and miscellaneous distracted activities). 3. Internal (i.e., residents' characteristics) and external (i.e., co-resident, direct caregiving, indirect caregiving, administrative, and government activities) influences affect residents' actions at mealtimes. The theory suggests that optimal mealtime experiences for residents require individualized care that reflects interdisciplinary, multi-level interventions. PMID- 15780151 TI - Does eating while watching television influence children's food-related behaviours? AB - To assess children's food-related behaviours and their relationships with eating while watching television (TV), data were collected from 534 ten-year-old French Canadian children. A self-administered questionnaire was used. Almost 18% of girls and over 25% of boys reported eating in front of the TV every day. Although, overall, the boys' eating pattern was less healthy than the girls', all of the children's food choices deteriorated with increased frequency of eating in front of the TV. Compared with girls, boys gave more importance to coloured and attractive foods, and to selecting foods similar to those eaten by others. Over 50% of children reported always receiving negative weight-related comments from family members. For boys, significant correlations were found between the frequency of eating in front of the TV, the importance given to a food's appearance, and their requests to parents for advertised foods. Significance was at the p<0.05 level for all findings. These results suggest that gender should be considered in attempts to understand children's food motivations and behaviours. The findings also indicate the need to document children's eating environments, and to inform children and their families about eating behaviours that may be associated with a given environment. PMID- 15780152 TI - Effect of gastrostomy on growth in children with neurodevelopmental disabilities. AB - Chronic malnutrition and growth failure are frequent consequences of feeding difficulties in neurodevelopmentally disabled children. Gastrostomy feeding has been used successfully to alleviate chronic malnutrition as well as distress and frustration associated with feeding. Unfortunately, caregivers are often resistant to gastrostomy placement. In order to determine the impact of gastrostomies in 20 children with neurodevelopmental disability (NDD), a questionnaire was used to collect caregivers' perceptions both before and after gastrostomy. The questionnaire assessed caregivers' retrospective perceptions of quality of life, feeding difficulties, and the burdens and benefits of gastrostomies. To determine impact on growth, height and weight were measured once before and three times after gastrostomy (at six, 12, and 24 months). The number of times a child was fed and the amount of time spent feeding decreased significantly following gastrostomy (p<0.001 and p<0.05, respectively). Growth for all children improved following gastrostomy (p<0.001). Pre-gastrostomy problems improved significantly following gastrostomy, as did caregivers' perceptions of quality of life for both themselves and their child (p<0.001). These results indicate that gastrostomy has a positive impact on growth for neurodevelopmentally disabled children, and on quality of life for both children and caregivers. Caregivers may find these results encouraging if they are faced with a decision about gastrostomy placement for their child. PMID- 15780153 TI - Factors that influence adherence to calcium recommendations. AB - To identify barriers to following calcium recommendations among women with reduced bone mineral density (BMD), four focus group sessions were held with 30 postmenopausal women diagnosed with low BMD. Key concepts and themes were derived from transcripts. Participants were aware of the importance of calcium to their bone health, and were attempting to follow calcium intake recommendations. Several major themes associated with the ability to obtain adequate calcium were identified, including knowledge and confidence in actions, lifestyle and food preferences, and side effects and conflict with other health conditions. Participants reported that they obtained information in an effort to make a confident decision about calcium intake, but were easily confused by conflicting information. Daily routines and family and personal food preferences influenced dietary behaviours. Women indicated that side effects, particularly those associated with perceived lactose intolerance, caused them to restrict their calcium intake. Our data provide important insight into factors that women believe affect their ability to reach recommended calcium intakes. To optimize osteoporosis prevention and treatment, dietitians should focus on individualized patient assessments to identify factors affecting adherence to dietary calcium recommendations. PMID- 15780154 TI - Media literacy: a critical role for dietetic practice. AB - The mass media, including broadcast, electronic, and print media, have become entrenched in Canadians' daily lives. Spending the majority of their leisure time with mass media puts Canadians at increased health risk. Our review of the research literature shows that television (TV) viewing and content are linked to potential consumer health risks due to developed health attitudes, beliefs, and behaviours. The associated health risks of children and youth are of particular concern. Excessive TV viewing has been associated with obesity development, increased energy consumption, reduced energy expenditure, negative body image development, and reduced concern with the concept of self-care. Media literacy, the ability to view critically and understand mediated messages, is a possible technique to mitigate these adverse effects. The enhanced inclusion of media literacy concepts in health education activities of dietetic practice is advocated. Dietitians could increase their understanding of research findings on the health-related effects of mass media use and the implications of including media literacy in daily practice. Such awareness would further augment available health promotion strategies. PMID- 15780155 TI - Meal composition and shift work performance. AB - Research indicates that the ability to perform a task can be affected by the composition of the meal preceding the task. This study investigated the effect of shift workers' consumption of a medium-fat, medium-carbohydrate meal on alertness scores. Six subjects (four men, two women) aged 19 to 44 recorded food intake, sleep, and quality of sleep for two weeks, and measured their body temperature and performed cognitive tests during two night shifts at baseline and in test periods. The Stanford Sleepiness Scale (SSS) was used to quantify sleepiness, and a Paced Auditory Serial Addition Test (PASAT) was used to measure cognitive performance. In comparison with the score at baseline, when subjects had a low fat, high-carbohydrate dietary intake (1,335 kcal/5,588 kJ, 56% carbohydrate, 28% fat), the 1.6-second PASAT score improved significantly (p=0.042) during night shifts when subjects consumed a test meal (987 kcal/4,131 kJ, 46% carbohydrate, 42% fat). No statistically significant difference in SSS was found between baseline and test periods. The reduced body temperature between 2400 hours and 0530 hours was similar for both baseline and test periods. Meal composition and size during night shifts may affect cognitive performance. PMID- 15780156 TI - Proceedings of the First Global Genito-Urinary Oncology Conference. October 2-3, 2004. Honolulu, Hawaii, USA. PMID- 15780157 TI - Detection of prostate cancer: the impact of the European Randomized Study of Screening for Prostate Cancer (ERSPC). AB - The European Randomized Study of Screening for Prostate Cancer (ERSPC) is a large, randomized controlled trial of screening versus control, conducted in eight European countries (Belgium, Finland, France, Italy, the Netherlands, Spain, Sweden, and Switzerland). This article focuses on important aspects relating to recent findings from the ERSPC about two topics: first, leadtime and overdiagnosis, and second, prostate-specific antigen (PSA) as a test for repeated screening. The ERSPC together with the prostate cancer arm of the Prostate, Lung, Colon and Ovary (PLCO) screening trial of the National Cancer Institute in the United States are set to show or exclude an effect of screening on prostate cancer mortality. Both studies are progressing according to plan. Definitive endpoint-related data can be expected between 2005 and 2010 depending on the difference in prostate cancer mortality that may be shown between the screening and control arms. The ERSPC will allow a risk-to-benefit analysis including parameters of quality of life and cost. Overdiagnosis with present prostate cancer screening regimens is high. This amount of overdiagnosis is likely to be unacceptable for most healthcare policy makers and providers. Addressing overdiagnosis will be a major research task for urologists for the years to come. Present screening needs to be more "selective" for cases that have aggressive patterns and are likely to lead to clinical diagnosis of prostate cancer and/or death. The test characteristics of prostate-specific antigen (PSA) change after one use. The positive relation between PSA levels and positive predictive value (PPV) and detection rates in first screening rounds are lost. This may be compatible with the observation that tumor volumes in second round screening are smaller, and larger tumors are harvested. Tumor volume becomes a negative predictor in round 2, indicating that a large proportion of elevated PSA values are caused by benign prostatic hyperplasia (BPH) rather than by prostate cancer. While the outcome of the ongoing randomized studies is uncertain, screening tests cannot be refused to men who are well-informed and accept to take the risk of experiencing more harm than benefit as a result of a positive screening test result. PMID- 15780158 TI - Screening for prostate cancer: updated experience from the Tyrol study. AB - OBJECTIVES: The aim of this study was to monitor the impact of prostate cancer screening in a natural experiment by comparing prostate cancer mortality in Tyrol, Austria, where prostate-specific antigen (PSA) testing was made available at no charge, with the rest of Austria, where this screening was not introduced. METHODS: In 1993, PSA testing was made freely available to men aged 40 to 79 years old living in the Federal State of Tyrol, Austria. In the first 10 years of this study, at least 70% of all men in this age range had PSA tests done at least once. Initially, only total PSA was measured, but free PSA measurement was added in 1995, and complexed PSA was added in 2001. Digital rectal examination (DRE) was not part of the screening examination. RESULTS: Significant migration to lower clinical as well as pathological prostate cancer stages has been observed in patients undergoing radical prostatectomy since the introduction of this screening program. A reduction in mortality rates from prostate cancer in Austria occurred from 1993 onward, with a much greater reduction in Tyrol; mortality remained fairly constant between 1993 and 1995 and subsequently fell. From 1993 to 2000 (the most recent data), there was a significantly greater decrease in the rate of prostate cancer mortality in Tyrol compared to the rest of Austria (P value = 0.006). Based on age-specific death rates for men aged 40 to 79, the difference between the number of expected and observed deaths from prostate cancer in Tyrol was 22 in 1998 (a 42% decrease), 18 in 1999 (a 33% decrease) and 25 in 2000 (a 44% decrease). CONCLUSIONS: These findings are consistent with the hypothesis that a policy of making PSA testing freely available, and wide acceptance by men in the population, is associated with a reduction in prostate cancer mortality in an area in which urology services and radiotherapy are available freely to all patients. It is our opinion that most of this decline in mortality is likely to be due to aggressive downstaging and successful treatment and that any contribution from detecting and treating early cancers will only become apparent in the years to come. PMID- 15780159 TI - Defining high-risk prostate cancer: current status. AB - Defining men at high risk for prostate cancer treatment failure and death continues to evolve. Identifying these men allows for better disease prognostication, patient decision treatment making and facilitates accrual for appropriate clinical trials. Men at traditional high risk for prostate cancer progression and death include men with advanced clinical stage, higher levels of PSA and Gleason pattern 4. Utilizing accepted methods of risk stratification including nomograms can aid in case identification. Softer risk factors such as obesity, race, socioeconomic status, and genetic polymorphisms are increasingly being studied. Ultimately high-throughput genomics will aid in identification of these men. PMID- 15780160 TI - Penile cancer: current challenges. AB - Penile cancer remains a formidable challenge in many developing countries because of its high incidence and the advanced disease stage at diagnosis. For early penile cancer, surgery alone offers a high cure rate. Penile sparing therapies are proposed as alternative treatment options for select patients with the added advantages of preservation of body image and improved quality of life. The optimal management of lymph node disease remains controversial. The role of the sentinel lymph node biopsy, lymphatic mapping, prophylactic lymphadenectomy and the template for lymph node dissection are discussed. For advanced, metastatic penile cancer, more effective and less toxic chemotherapy is needed. This may be coupled with palliative surgery or radiotherapy for the primary tumor and inguinal disease. PMID- 15780161 TI - Sentinel lymph node biopsy in penile cancer: evolution and insights. AB - INTRODUCTION: Defining the presence or absence of microscopic metastases in the inguinal lymph nodes in patients with invasive penile squamous carcinoma and no palpable adenopathy remains crucial but difficult short of performing inguinal lymphadenectomy. METHODS: We reviewed the results of less invasive procedures such as traditional sentinel node biopsy and contemporary dynamic sentinel node biopsy aided by intraoperative lymphatic mapping (IOLM) to determine their role in the management of patients without palpable inguinal adenopathy RESULTS: Inguinal node biopsy directed to the sentinel node area or region although initially promising was associated with a recurrence rate of 16% (24/150) among seven series reported. Extending the dissection to a wider region did not improve these results (20% recurrence, 5/25 patients). Preoperative lymphoscintigraphy combined with IOLM (with blue dye and a hand held gamma probe to detect radioactive counts) routinely detected sentinel nodes in the Netherlands Cancer Institute experience. However IOLM was associated with a false negative rate of 18% (6/34 patients). IOLM using an open incision approach at M.D. Anderson Cancer Center provided evidence for inguinal lymph drainage to alternate areas within the inguinal field confirming proof of principle for IOLM. CONCLUSION: Inguinal lymph node biopsy directed to the sentinel node area to detect microscopic metastases is no longer recommended. Dynamic sentinel node biopsy utilizing IOLM is a promising technique in evolution that requires further testing among high volume centers for penile cancer. Contemporary superficial and modified inguinal dissection techniques with intraoperative frozen section remain the "gold standard" for defining the presence of microscopic metastases. PMID- 15780162 TI - Extended lymphadenectomy in penile cancer. AB - INTRODUCTION AND OBJECTIVE: There are many controversies regarding the optimal management of the inguinal nodes in patients with penile cancer. The inflammatory response of the draining regional lymph nodes can cause enlargement without implying the presence of metastases. On the other hand, 20% of patients with clinically non-suspicious nodes contain micrometastases. We studied the dissemination risk factors of the primary lesion in penile cancer, the preferential lymphatic pathways, and the extension of lymphadenectomies, in order to understand how to better control this cancer. PATIENTS AND METHODS: In this prospective study of 50 patients (aged 21-73; median age 54) with penile carcinoma, the initial clinical and pathologic findings were compared to biopsy specimens obtained in routine, bilateral, superficial, deep and pelvic lymphadenectomies. Patients were followed from 6 months to 18 years. RESULTS: We found that first, risk factors of tumor spread were primary lesions greater than 2 cm in diameter, unfavorable histology findings, and invasive lesions. Second, these risk factors were present in all patients who had node metastases. Third, clinical staging was not accurate, since there was a tendency to understage 19% of localized disease and overstage 51.5% of metastatic disease. Fourth, the sentinel nodes were the most commonly infiltrated nodes but were only present in 2 (11%) of 18 metastatic cases. Fifth, there were no cases of deep inguinal nodes without superficial infiltration. Lastly, there were no cases of pelvic node without prior contamination of inguinal nodes. CONCLUSIONS: Ideal candidates for watchful waiting after primary lesion treatment are those who do not have primary lesions greater than 2 cm in diameter, unfavorable histology findings, invasive lesions, or palpable nodes. Performing limited surgery on positive nodes risks leaving some of the tumor. Superficial lymphadenectomy is the procedure of choice in cases of patients with clinical negative nodes and risk factors of tumor spread. PMID- 15780163 TI - Open retroperitoneal lymph node dissection. AB - Testicular cancer has become one of the most curable solid malignancies. Although chemotherapy can rescue patients with advanced disease, accurate staging of the retroperitoneum has been a mainstay of effectiveness of treatment. Retroperitoneal lymph node dissection via open technique has been and remains the gold standard for pathologic staging of the retroperitoneum as well as effective therapy for patients with minimal nodal involvement. Retroperitoneal lymphadenectomy has resulted in a 99.5% tumor survival for patients with clinical stage I or early stage II disease. Alternatives to open retroperitoneal lymph node dissection include laparoscopic retroperitoneal lymph node dissection, active surveillance, or primary chemotherapy. Each of these modalities has a potential role in selected patients. For the majority of patients, however, a meticulous retroperitoneal lymph node dissection gives patients the highest likelihood of survival with relatively low morbidity. PMID- 15780164 TI - UPM3: review of a new molecular diagnostic urine test for prostate cancer. AB - PSA elevation is the most common indication for urologic referral to rule out the presence of prostate cancer. Recently PSA screening and its usefulness in suggesting the presence of clinically significant prostate cancer has been put into doubt. PSA has limitations in detecting significant cancers even when elevated and on the other hand significant cancers are found in the presence of low PSA levels. In order to better predict patients at risk of harboring prostate cancer new diagnostic tests are required. A promising novel approach is based on the molecular detection of prostate cancer cells in urine. The uPM3 assay is based on the amplification of specific target RNA using the nucleic acid sequence based amplification (NASBA) technology. In a large multi-center study of 517 cases the overall sensitivity was 66% with a specificity of 89%. The positive (PPV) predictive values for the uPM3 test were 75% compared to 38% for a serum PSA cutoff of 4. The negative predictive value (NPV) was equivalent between the tests, but due to the higher PPV for uPM3, the accuracy of uPM3 was nearly two fold greater than PSA (81% versus 43% and 47% for PSA cutoffs of 2.5 and 4 ng/ml, respectively). This test may become one of the first molecular diagnostic tools to aid in prostate cancer detection. PMID- 15780165 TI - Prostate biopsy: who, how and when. An update. AB - Biochemical parameters and pathological features as well as biopsy related morbidity of prostate cancer detected on second, third and fourth repeat prostate biopsy in men with a serum total PSA level between 4 ng/mL and 10 ng/mL were evaluated and compared to those cancers detected on initial prostate biopsy. In a prospective European Prostate Cancer Detection study, 1051 men with a total PSA level between 4 ng/mL and 10 ng/mL underwent transrectal ultrasound (TRUS)-guided sextant biopsy and two additional transition zone biopsies. All subjects whose biopsy samples were negative for prostate cancer (CaP) underwent a first repeat biopsy after 6 weeks. If also negative a third and even a fourth biopsy was performed at 8 weeks intervals. Those with clinically localized cancers underwent radical prostatectomy. Pathological and clinical features of patients diagnosed with cancer on either initial or repeat biopsy and clinically organ confined disease who agreed to undergo radical prostatectomy were compared. Cancer detection rates on first, second, third and fourth biopsy were 22% (231/1051), 10% (83/820), 5% (36/737) and 4% (4/94), respectively. Percent free PSA and PSA TZ were the most powerful parameters to predict cancer on repeat biopsy. Overall, of patients with clinically localized disease (67% of cancers detected), 86% underwent radical prostatectomy and 14% opted for watchful waiting or radiation therapy. Overall, 58.0%, 60.9%, 86.3% and 100% had organ confined disease on first, repeat, third and fourth biopsy, respectively. Despite statistical significant differences with respect to multifocality (p=0.009) and cancer location (p=0.001) (cancers on second biopsy showing a lower rate of multifocality and a more apico-dorsal location), there were no differences with respect to stage (p=0.2), Gleason score (p=0.3), percentage Gleason grade 4/5 (p=0.2), serum PSA and patient age between first and second biopsy. However, cancers detected on third and fourth biopsy had a significantly lower Gleason score (p=0.001 and 0.001), lower rate of grade 4/5 cancer (p=0.02), lower cancer volume (p= 0.001 and 0.001) and lower stage (p= 0.001). Morbidity of first and repeat biopsy were similar, whereas third and fourth biopsy had a slightly higher complication rate. Interestingly, patients under 60 years of age reported a higher pain apprehension as quantified with the visual analog pain scale (VAS). Further, the use of the Vienna tables allowed an accurate calculation of the number of biopsy cores required based on prostate volume and age. Despite differences in location and multifocality, pathological and biochemical features of cancers detected on initial and second biopsy were similar, suggesting similar biological behavior. Cancers detected on third and fourth biopsy had a lower grade, stage and cancer volume as compared to cancers on first and repeat biopsy. Morbidity of first and repeat biopsy were similar, whereas third and fourth biopsy had a slightly higher complication rate. Hence, a second prostate biopsy in all cases of a negative finding on initial biopsy appears justified. Third and fourth repeat biopsies however, should only be obtained in very selected patients with high suspicion of cancer and/or poor prognostic factors on the first or second biopsy. Power Doppler TRUS will further enhance prostate cancer detection as will artificial neural networks as patient selecting tools. PMID- 15780166 TI - Prostatic intraepithelial neoplasia: a risk factor for prostate cancer. AB - Prostatic Intraepithelial Neoplasia (PIN) is an increasingly common finding at ultrasound guided prostate biopsy, with the high grade form (HGPIN) thought to be "precancerous". With the more widespread use of extended biopsy protocols, taking sometimes up to 14 cores or more, the incidence of HGPIN can be up to 25%. Histologically, it has many features in common with cancer of the prostate and has been shown to be both associated with cancer at the time of its finding and predictive for the development of prostate cancer in the future. Basic science research has demonstrated genes common specifically to both prostate cancer and HGPIN and immunostaining studies of microvessel density may help to differentiate HGPIN from lower risk PIN. There are no active treatments for HGPIN although there are trials to assess the effectiveness of hormonal therapy and nutritional supplements. Currently most urologists recommend that patients should be followed at 6 monthly intervals with regular PSA and repeat biopsies as indicated. PMID- 15780167 TI - Active surveillance with selective delayed intervention: walking the line between overtreatment for indolent disease and undertreatment for aggressive disease. AB - PURPOSE: To summarize the case for active surveillance for good risk prostate cancer with selective delayed intervention for rapid biochemical or grade progression, and review the results of a large phase II experience using this approach. MATERIALS AND METHODS: A prospective phase II study of active surveillance with selective delayed intervention was initiated in 1995. Patients were managed initially with surveillance; those who had a PSA DT of 2 years or less, or grade progression on rebiopsy were offered radical intervention. The remainder were closely monitored. RESULTS: The cohort consists of 299 patients with good risk prostate cancer, or intermediate risk prostate cancer in men over 70. The median PSA doubling time was 7.0 years. 35% had a PSA DT > 10 years. The majority of patients remain on surveillance. At 8 years, overall actuarial survival is 85%, and disease specific survival is 99%. CONCLUSION: Most men with favorable risk prostate cancer will die of unrelated causes. The approach of active surveillance with selective delayed intervention based on PSA DT represents a practical compromise between radical therapy for all (which results in overtreatment for patients with indolent disease), and watchful waiting with palliative therapy only (which results in undertreatment for those with aggressive disease). The results at 8 years are favorable. Longer follow up will be required to confirm the safety of this approach in men with a long (>15 year) life expectancy. PMID- 15780168 TI - Surgery or radiation: what is the optimal management for locally advanced prostate cancer? AB - INTRODUCTION: To date, randomized trials comparing radiotherapy to surgery for adenocarcinoma of the prostate are few. Lacking, are randomized comparisons between treatment modalities for the patient with high-risk locally advanced disease. Hence, there is a need to determine which approach offers superior results in these patients who comprise a significant proportion of those dying of prostate cancer. In this short review we highlight key studies that may provide interim answers while awaiting definitive results from randomized studies. MATERIAL AND METHODS: A MEDLINE literature review was performed of studies evaluating current treatment modalities for high-risk (TNM stage >T2b, PSA>10, Gleason > or =8) prostate cancer. Publications from 1975 to present were searched using the keywords: prostate cancer, locally advanced prostate cancer, high-risk prostate cancer, prostatectomy, external beam radiation, brachytherapy, and PSA doubling time. RESULTS: Comparisons of different treatment modalities are difficult due to many factors, from uncertainties in clinical staging to the questionable equivalence of PSA failure. However, the general consensus is that low dose rate brachytherapy monotherapy is not ideal for high-risk patients. There are several options for combination therapy which show moderately good survival results. Because of the lack of prospective randomized trials comparing these approaches, matched analyses with uniform patient treatment and pathological review may provide an interim answer. CONCLUSION: The optimal management for patients with locally advanced prostate cancer is unclear. While randomized clinical trials will eventually shed light on this question, interim solutions may provide some answers in the short term. PMID- 15780169 TI - Stage I nonseminomatous germ cell tumors: the case for management by risk stratification. AB - For patients with clinical stage I nonseminomatous germ cell tumor (NSGCT), the therapeutic options after orchiectomy are retroperitoneal lymphadenectomy, surveillance, and chemotherapy. Ideally the option selected will be based on an individualized assessment of the estimated risk of progression based on prognostic factors, so called risk-adapted treatment, to reduce overall burden of therapy while maintaining survival. It is possible to identify patients at low risk of progression who can be followed by active surveillance initially. Prognostic factors for high risk, while well defined, do not identify all patients at risk and those that are destined to progress, can usually be salvaged with delayed therapy. Most centres recommend either surgery or primary chemotherapy for those defined as being at high risk for progression. Prognostic factors for high risk however, while well defined, do not identify all patients at risk and those that are destined to progress, can usually be salvaged with delayed therapy. PMID- 15780170 TI - New treatments for metastatic kidney cancer. AB - Renal cell carcinoma accounts for approximately 3% of adult malignancies and 90% 95% of neoplasms arising from the kidney. It is characterized by a lack of early warning signs, diverse clinical manifestations, resistance to radiation and chemotherapy, and infrequent but reproducible responses to immunotherapy with agents such as interferon alpha (IFNa) and interleukin 2 (IL-2). International studies have shown objective response rates of < 15% in patients with advanced and metastatic disease, with 5-year disease-specific survival ranging between 0 20%. Considering these poor outcomes, renal cancers' very vascular nature and overexpression of receptors for vascular endothelial growth factor (VEGF), various biologic and angio-suppressive therapies are being evaluated in clinical trials. Promising results in terms of overall response rate and median time to progression have been reported especially as second-line therapy following cytokine failure, a setting where no effective systemic therapy has been recognized (SU011248, Bay 43-9006, Bevacizumab and Erlotinib). While confirmatory studies are ongoing, other novel treatments in first line trials (CCI-779, Infliximab, PTK-787, and Thalidomide) have drawn international attention. This review, analyzing basic translational research principles, will summarize the available data on the use of these new therapeutic approaches in RCC. PMID- 15780171 TI - Bone loss in prostate cancer: evaluation, treatment and prevention. AB - Modern medicine offers multiple treatment options to prolong the survival of patients with prostate cancer. However, in the absence of adequate supportive care, the systemic effects of prostate cancer and therapies such as androgen deprivation therapy (ADT) can undermine skeletal integrity, resulting in skeletal complications. Skeletal morbidity contributes to the erosion in quality of life in patients with prostate cancer. These patients are at risk for fractures from cancer treatment-induced bone loss and, later on, pathologic fractures from bone metastases, which may occur during the progression of prostate cancer. Several supportive care options are available to prevent generalized and focal bone loss, including calcium and vitamin D supplements and bisphosphonates. Oral calcium and vitamin D supplementation alone, however, appears to be insufficient to prevent bone loss during ADT. Bisphosphonates may be beneficial in preventing bone loss and eventually reducing skeletal morbidity due to prostate cancer and ADT. PMID- 15780172 TI - Effectiveness of Maximal Androgen Blockade (MAB): illusion or reality? AB - Two decades have passed since the concept of Maximal Androgen Blockade (MAB) was first applied to the clinical treatment of prostate cancer. The theory is that by cutting off the supply of androgen from the adrenal gland, androgen blockade of the prostate could be made more complete. However, to date the clinical benefit of MAB has failed to live up to the theoretically expected effect. Having said that, fundamental research and clinical trials in recent years do indicate that the benefit of MAB is not merely an illusion. PMID- 15780173 TI - Recent docetaxel studies establish a new standard of care in hormone refractory prostate cancer. AB - INTRODUCTION: Treatment of hormone refractory prostate cancer (HRPC) has generally aimed at increasing symptom free survival in asymptomatic patients and improving quality of life in symptomatic patients. However, recent randomized studies might be shifting the paradigm towards achieving an improved overall survival. METHODS: Two large randomized controlled studies were conducted using mitoxantrone plus prednisone as a control arm compared to docetaxel-based regimens. RESULTS: In the TAX 327 trial, 3-weekly docetaxel plus prednisone proved significantly superior to mitoxantrone plus prednisone (an established reference regimen) in extending survival, reducing levels of prostate specific antigen (PSA), controlling pain and improving quality of life. The Southwest Oncology Group's study (protocol 99-16) randomized patients to either docetaxel plus estramustine or mitoxantrone plus prednisone. Compared with the mitoxantrone regimen, docetaxel plus estramustine significantly extended median overall survival and time to progression. Men treated with the docetaxel regimen were also more likely to have a PSA response. In this study, the two regimens were similarly effective in relieving pain. CONCLUSION: These studies have an important impact on the management strategy of hormone refractory prostate cancer. Docetaxel is the first agent shown significantly to extend survival in HRPC. Although this proven benefit must be balanced against toxicity, docetaxel should now be considered the standard of care for most patients that fail first line or more hormonal manipulations. Drug combinations which may further extend survival and improve quality of life are actively being pursued, as is the possibility of using docetaxel in the adjuvant setting. PMID- 15780174 TI - International regional working groups on prostate cancer: results of consensus development. AB - At the first Global Urologic Oncology Congress--which was held in conjunction with the SIU meeting in Honolulu, Hawaii, on October 2-3, 2004--a total of 250 urologists from nine regions throughout the world participated in working groups to develop consensus statements about the management of prostate cancer patients. The focus was on two areas of prostate cancer: first, screening and detection- including when to do a biopsy, and how to manage low-risk patients--and second, management of high-risk patients. CONCLUSION: Overall, there is marked global diversity of beliefs about optimal screening and management of patients with low- or high-risk localized prostate cancer. This diversity likely reflects regional differences in epidemiologic factors, resources, and treatment patterns. Participating in the process of trying to develop international practice consensus gives urologists the opportunity to broaden their perspective about handling common clinical scenarios. PMID- 15780175 TI - Viral infections and cell cycle G2/M regulation. AB - Progression of cells from G2 phase of the cell cycle to mitosis is a tightly regulated cellular process that requires activation of the Cdc2 kinase, which determines onset of mitosis in all eukaryotic cells. In both human and fission yeast (Schizosaccharomyces pombe) cells, the activity of Cdc2 is regulated in part by the phosphorylation status of tyrosine 15 (Tyr15) on Cdc2, which is phosphorylated by Wee1 kinase during late G2 and is rapidly dephosphorylated by the Cdc25 tyrosine phosphatase to trigger entry into mitosis. These Cdc2 regulators are the downstream targets of two well-characterized G2/M checkpoint pathways which prevent cells from entering mitosis when cellular DNA is damaged or when DNA replication is inhibited. Increasing evidence suggests that Cdc2 is also commonly targeted by viral proteins, which modulate host cell cycle machinery to benefit viral survival or replication. In this review, we describe the effect of viral protein R (Vpr) encoded by human immunodeficiency virus type 1 (HIV-1) on cell cycle G2/M regulation. Based on our current knowledge about this viral effect, we hypothesize that Vpr induces cell cycle G2 arrest through a mechanism that is to some extent different from the classic G2/M checkpoints. One the unique features distinguishing Vpr-induced G2 arrest from the classic checkpoints is the role of phosphatase 2A (PP2A) in Vpr-induced G2 arrest. Interestingly, PP2A is targeted by a number of other viral proteins including SV40 small T antigen, polyomavirus T antigen, HTLV Tax and adenovirus E4orf4. Thus an in-depth understanding of the molecular mechanisms underlying Vpr-induced G2 arrest will provide additional insights into the basic biology of cell cycle G2/M regulation and into the biological significance of this effect during host pathogen interactions. PMID- 15780176 TI - The matrix metalloproteinase stromelysin-3 cleaves laminin receptor at two distinct sites between the transmembrane domain and laminin binding sequence within the extracellular domain. AB - The matrix metalloproteinase (MMP) stromelysin-3 (ST3) has long been implicated to play an important role in extracellular matrix (ECM) remodeling and cell fate determination during normal and pathological processes. However, like other MMPs, the molecular basis of ST3 function in vivo remains unclear due to the lack of information on its physiological substrates. Furthermore, ST3 has only weak activities toward all tested ECM proteins. Using thyroid hormone-dependent Xenopus laevis metamorphosis as a model, we demonstrated previously that ST3 is important for apoptosis and tissue morphogenesis during intestinal remodeling. Here, we used yeast two-hybrid screen with mRNAs from metamorphosing tadpoles to identify potential substrate of ST3 during development. We thus isolated the 37 kd laminin receptor precursor (LR). We showed that LR binds to ST3 in vitro and can be cleaved by ST3 at two sites, distinct from where other MMPs cleave. Through peptide sequencing, we determined that the two cleavage sites are in the extracellular domain between the transmembrane domain and laminin binding sequence. Furthermore, we demonstrated that these cleavage sites are conserved in human LR. These results together with high levels of human LR and ST3 expression in carcinomas suggest that LR is a likely in vivo substrate of ST3 and that its cleavage by ST3 may alter cell-extracellular matrix interaction, thus, playing a role in mediating the effects of ST3 on cell fate and behavior observed during development and pathogenesis. PMID- 15780177 TI - Overexpression of Bax sensitizes prostate cancer cells to TGF-beta induced apoptosis. AB - NRP-154 is a tumorigenic epithelial cell line derived from the preneoplastic dorsal-lateral prostate of rats. These cells are exquisitely sensitive to TGF beta induced apoptosis. In contrast, we find that NRP-154 cells can sustain overexpression of exogenous Bax protein, which is different from non-tumor cells where Bax functions as a ubiquitous stimulator of apoptosis. NRP-154 cells stably overexpressing Bax show increased sensitivity to TGF-beta induced apoptosis. The degree of TGF-beta induced apoptosis displays high correlation with cleavage of Bax at the amino-terminus. Our data indicate that prostate cancer cells can host high levels of latent Bax which can be activated through post-translational modification. PMID- 15780179 TI - Residues Met76 and Gln79 in HLA-G alpha1 domain involve in KIR2DL4 recognition. AB - Human leukocyte antigen-G (HLA-G) has long been speculated as a beneficial factor for a successful pregnancy for its restricted expression on fetal-maternal extravillous cytotrophoblasts and its capability of modulating uterine natural killer cell (uNK) function such as cytotoxicity and cytokine production through NK cell receptors. HLA class I alpha1 domain is an important killer cell Ig-like receptor (KIR) recognition site and the Met76 and Gln79 are unique to HLA-G in this region. NK cell receptor KIR2DL4 is a specific receptor for HLA-G, yet the recognition site on HLA-G remains unknown. In this study, retroviral transduction was applied to express the wild type HLA-G (HLA-wtG), mutant HLA-G (HLA-mG) on the chronic myelogenous leukemia cell line K562 cells and KIR2DL4 molecule on NK 92 cells, respectively. KIR2DL4-IgG Fc fusion protein was generated to determine the binding specificity between KIR2DL4 and HLA-G. Our results showed that residue Met76, Gln79 mutated to Ala76,79 in the alpha1 domain of HLA-G protein could affect the binding affinity between KIR2DL4 and HLA-G, meanwhile, the KIR2DL4 transfected NK-92 cells (NK-92-2DL4) showed a considerably different cytolysis ability against the HLA-wtG and HLA-mG transfected K562 targets. Taken together, our data indicated that residue Met76 and Gln79 in HLA-G alpha1 domain plays a critical role in the recognition of KIR2DL4, which could be an explanation for the isoforms of HLA-G, all containing the a1 domain, with the potential to regulate NK functions. PMID- 15780178 TI - Indoleamine 2, 3-dioxygenase (IDO) is essential for dendritic cell activation and chemotactic responsiveness to chemokines. AB - Indoleamine 2, 3-dioxygenase (IDO) is a rate-limiting enzyme for the tryptophan catabolism. In human and murine cells, IDO inhibits antigen-specific T cell proliferation in vitro and suppresses T cell responses to fetal alloantigens during murine pregnancy. In mice, IDO expression is an inducible feature of specific subsets of dendritic cells (DCs), and is important for T cell regulatory properties. However, the effect of IDO and tryptophan deprivation on DC functions remains unknown. We report here that when tryptophan utilization was prevented by a pharmacological inhibitor of IDO, 1-methyl tryptophan (1MT), DC activation induced by pathogenic stimulus lipopolysaccharide (LPS) or inflammatory cytokine TNF-alpha was inhibited both phenotypically and functionally. Such an effect was less remarkable when DC was stimulated by a physiological stimulus, CD40 ligand. Tryptophan deprivation during DC activation also regulated the expression of CCR5 and CXCR4, as well as DC responsiveness to chemokines. These results suggest that tryptophan usage in the microenvironment is essential for DC maturation, and may also play a role in the regulation of DC migratory behaviors. PMID- 15780180 TI - Golgi localization and dynamics of hyaluronan binding protein 1 (HABP1/p32/C1QBP) during the cell cycle. AB - Hyaluronan binding protein 1 (HABP1) is a negatively charged multifunctional mammalian protein with a unique structural fold. Despite the fact that HABP1 possesses mitochondrial localization signal, it has also been localized to other cellular compartments. Using indirect immunofluorescence, we examined the sub cellular localization of HABP1 and its dynamics during mitosis. We wanted to determine whether it distributes in any distinctive manner after mitotic nuclear envelope disassembly or is dispersed randomly throughout the cell. Our results reveal the golgi localization of HABP1 and demonstrate its complete dispersion throughout the cell during mitosis. This distinctive distribution pattern of HABP1 during mitosis resembles its ligand hyaluronan, suggesting that in concert with each other the two molecules play critical roles in this dynamic process. PMID- 15780181 TI - Preliminary studies on differential defense responses induced during plant communication. AB - We compared the expression patterns of three representative genes in undamaged tomato and tobacco plants in response to exposure to either tomato or tobacco fed on by Helicoverpa armigera (cotton bollworm). When tomato and tobacco, two species of one family, were incubated in the chambers with the tomato plants damaged by the cotton bollworm, the expression of the PR1, BGL2, and PAL genes was up-regulated in leaves of both plants. However, the levels of gene expression were significantly higher in the tomato than that in the tobacco. In addition, the activities of enzymes, peroxidase, polyphenol oxidase, and lipoxygenase were found to be higher in the tomato than those in the tobacco. Similar results were obtained when the damaged plants were replaced by the tobacco. PMID- 15780182 TI - Inhibition of genes expression of SARS coronavirus by synthetic small interfering RNAs. AB - RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequence. dsRNA-mediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression. Synthetic 21-23 nucleotide (nt) small interfering RNA (siRNA) with 2 nt 3' overhangs was recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we studied the effects of synthetic siRNA duplexes targeted to SARS coronavirus structural proteins E, M, and N in a cell culture system. Among total 26 siRNA duplexes, we obtained 3 siRNA duplexes which could sequence-specifically reduce target genes expression over 80% at the concentration of 60 nM in Vero E6 cells. The downregulation effect was in correlation with the concentrations of the siRNA duplexes in a range of 0 approximately 60 nM. Our results also showed that many inactive siRNA duplexes may be brought to life simply by unpairing the 5'end of the antisense strands. Results suggest that siRNA is capable of inhibiting SARS coronavirus genes expression and thus may be a new therapeutic strategy for treatment of SARS. PMID- 15780183 TI - Endostatin promotes the anabolic program of rabbit chondrocyte. AB - Endostatin is a natural occurred angiogenesis inhibitor derived from collagenXVIII. So far its function during the angiogenesis process of bone formation and arthropathy has not been well studied yet. The present study addresses the function of endostatin in rabbit articular chondrocytes (RAC). We found that endostatin can promote RAC adhesion and spreading as well as its proliferation. In monolayer cultured RAC, CollagenII, TIMP1 and collagenXVIII transcription were up regulated by endostatin while collagenI and MMP9 were down regulated. Moreover collagenXVIII and endostatin antigens are present at synovial fluid. These findings indicate new function of endostatin as a homeostatic factor in cartilage metabolism. PMID- 15780184 TI - Identification of connexin 50 and 57 mRNA in A-type horizontal cells of the rabbit retina. AB - Horizontal cells (HCs) mediate negative feedback to photoreceptors. In the mammalian retina, there are two types of HCs, which are extensively coupled to neighboring cells through homologous gap junctions. The permeability and therefore the strength of feedback can be regulated by light intensity, dopamine and many other factors. However, the component(s) of the most prominent gap junctions, those between A-type HCs in the rabbit retina, is still unknown. In this study, we compared the sequences of many types of mammalian connexins, obtained partial sequences of rabbit connexin 50 and 57. Using specific primers designed against the rabbit sequences, we identified mRNAs of connexin 50 and/or 57 in visually selected single A-type HC using multiplex RT-PCR. PMID- 15780185 TI - Identification of EGFR kinase domain mutations among lung cancer patients in China: implication for targeted cancer therapy. AB - Lung cancer is one of the leading causes of death with one of the lowest survival rates. However, a subset of lung cancer patients who are of Asian origin and carry somatic mutations in epidermal growth factor receptor or EGFR have responded remarkable well to two tyrosine kinase inhibitors, gefitinib and erlotinib. While EGFR mutation profiles have been reported from Japan, South Korea, and Taiwan, there is no such report from mainland of China where the largest pool of patients reside. In this report, we identified ten somatic mutations from a total of 41 lung cancer patients in China. Among them, seven mutations were found in 17 adenocarcinomas. In contrast to previous reports, eight of these mutations are deletions in exon 19 and two of these deletions are homozygous. These results suggest that a large portion of Chinese adenocarcinoma patients could benefit from gefitinib or erlotinib. This unique mutation profile provides a rationale to develop the next generation of EGFR inhibitors more suitable for the Chinese population. PMID- 15780186 TI - Role of amygdala in mediating sexual and emotional behavior via coupled nitric oxide release. AB - Although the anatomical configuration of the amygdala has been studied a great deal, very little research has been conducted on understanding the precise mechanism by which this emotional regulatory center exerts its control on emotional and sexual behavior. By applying research methodology from the Neuroscience Research Institute, State University of New York, College at Old Westbury, we intended to demonstrate that much of the mediated effects of the amygdala, specifically the regulation of the male and female sexual response cycles, as well as related emotional considerations, exert their effects coupled to nitric oxide (NO) release. Furthermore, by using current anatomical and histological data, we demonstrated that amygdalar tissue rich in endocannabinoid and opiate, as well as catecholamine, receptors could exert its neurochemical effects within an NO-mediated paradigm. This paradigm, together with the existence of estrogen and androgen signaling within the amygdala, further lends credence to our theoretical framework. We begin with a brief anatomical and functional review of amygdalar function, and then proceed to demonstrate its relationship with NO. PMID- 15780187 TI - Development of Caenorhabditis elegans pharynx, with emphasis on its nervous system. AB - The Caenorhabditis elegans pharynx is a neuromuscular tube of which the function is to pump and crush bacteria, and inject them into the intestine. The 80-cell pharynx develops via the morphogenesis and differentiation of the cells that compose its semi-spherical primordium, and requires the activity of several evolutionarily conserved genes, such as pha-4 (the homolog to the Drosophila forkhead and vertebrate FoxA), ceh-22 (the homolog to the Drosophila tinman and vertebrate Nkx2.5), and pha-2 (the homolog to the vertebrate Hex). There are 20 neurons in the pharynx, each with a reproducible unique trajectory. Developmental genetic analysis of axon guidance in the pharynx indicates that some axon trajectories are in part established without growth cones, whereas other parts necessitate growth cone function and guidance. Here we provide an overview of the developmental genetics of the Caenorhabditis elegans pharynx, with an emphasis on its nervous system. PMID- 15780188 TI - Baculovirus as a highly efficient expression vector in insect and mammalian cells. AB - Baculovirus has been widely used for the production of recombinant proteins in insect cells. Since the finding that baculovirus can efficiently transduce mammalian cells, the applications of baculovirus have been greatly expanded. The prospects and drawbacks of baculovirus-mediated gene expression, either in insect or in mammalian cells, are reviewed. Recent progresses in expanding the applications to studies of gene regulation, viral vector preparation, in vivo and ex vivo gene therapy studies, generation of vaccine vectors, etc are discussed and the efforts directed towards overcoming the existing bottlenecks are particularly emphasized. PMID- 15780189 TI - Novel role for gabapentin in neuroprotection of central nervous system in streptozotocine-induced diabetic rats. AB - AIM: To investigate the effect of gabapentin on neural [neuron-specific enolase (NSE)] and glial markers [glial fibrillary acidic protein (GFAP) and S100B] in different brain regions of diabetic rats. METHODS: Diabetes was induced by a single intraperitoneal injection of streptozotocine (50 mg/kg body weight). Rats in one diabetic group received gabapentin (50 mg.kg(-1).d(-1)) and rats in the other diabetic group received vehicle only for 6 weeks. The levels of GFAP, S100B, and NSE were determined by immunoblotting in the hippocampus, cortex, and cerebellum. Lipid peroxidation (LPO as malondialdehyde+ 4-hydroxyalkenals) and glutathione (GSH) levels were also determined in the same brain parts. RESULTS: Total and degraded GFAP content and S100B protein expression in different areas of brain tissues significantly increased in diabetic rats compared to control rats. Similarly, NSE levels were also significantly elevated in hyperglycemic rats. In addition, there was a significant increase in LPO levels in the diabetic rat brain compared to control rat brains. Pretreatment with gabapentin prevented the upregulation of GFAP, S100B, and NSE in all brain regions of diabetic rats. The level of LPO was reduced, but not completely halted, by treatment with gabapentin. CONCLUSION: These results suggest that diabetes causes glial and neuronal injury, possibly as a result of elevated oxidative stress, and that gabapentin protects neurons and glial cells. Thus, we predict that gabapentin treatment will attenuate the hippo-campal and cortical neurodegeneration observed during diabetes mellitus in rats. PMID- 15780190 TI - Influence of low dietary histamine on seizure development of chemical kindling induced by pentylenetetrazol in rats. AB - AIM: To determine the role of dietary low histamine on the seizure development of pentylenetetrazol (PTZ)-induced kindling in rats. METHODS: After 14 d of feeding on a low histamine diet (LH, containing 0.145 mumol/g of histamine), the rats were chemically kindled by repeated intraperitoneal injection of a subconvulsant dose of PTZ (35 mg/kg) once every 48 h, and seizure activity of kindling was recorded for 30 min. Histamine in brain samples was analyzed using a high performance liquid chromatography system with a fluorescence spectrofluorometer. RESULTS: The LH diet induced an increase in seizure response (seizure susceptibility) to the first trial of PTZ, and resulted in facilitation of subsequent PTZ kindling process (seizure development). The histamine levels in the cortex, hippocampus, and hypothalamus of LH-treated rats decreased significantly and these changes correlated well with seizure behavior (r = 0.875, 0.651, and 0.796, respectively). In addition, chronic kindled seizures resulted in a significant increase of the histamine content in the cortex and hypothalamus in the LH-fed groups. CONCLUSION: These findings indicate that the histamine in daily food could influence the brain histaminergic function, and play an important role in regulating seizure susceptibility. PMID- 15780191 TI - Bradykinin potentiates 5-HT3 receptor-mediated current in rat trigeminal ganglion neurons. AB - AIM: To explore the modulatory effect of bradykinin (BK) on 5-HT(3 )receptor mediated current in trigeminal ganglion (TG) neurons in rats. METHODS: The whole cell patch-clamp technique was used to record 5-HT-activated currents (I(5-HT)) in neurons freshly dissociated from rat TG. Drugs were applied by rapid solution exchange. RESULTS: The majority of the neurons examined responded to 5-HT applied externally with an inward current (76.3%, 74/97) that could be blocked by the 5 HT(3 )receptor antagonist, ICS-205,930 (10(-6) mol/L). In 66 of the 74 cells sensitive to 5-HT (89.2%), pretreatment for 30 s with BK (10(-6)-10(-10) mol/L) could potentiate I(5-HT) with the maximal modulatory effect occurring at 10(-7) mol/L BK (71.6%+/-4.9%). BK shifted the 5-HT concentration-response curve upwards with an increase of 68.9%+/-7.2% in the maximal current response, but with no significant change in the EC(50) value (19.1+/-3.2 mumol/L vs 20.9+/-3.5 micromol/L; t-test, P>0.05; n=8). BK potentiated I(5-HT) in a holding potential independent manner and did not alter the reverse potential of I(5-HT). This BK induced potentiation of I(5-HT) was almost completely blocked by Hoe 140 (5*10( 7) mol/L), a selective B2 BK receptor antagonist, and was removed after intracellular dialysis of GF-109203X (2 micromol/L), a selective protein kinase C (PKC) inhibitor, with the re-patch clamp. CONCLUSION: Pre-application of BK exerts an enhancing effect on I(5-HT) via a PKC-dependent pathway in rat TG neurons, which may explain the peripheral mechanism of pain and hyperalgesia caused by, for example, tissue damage and inflammation. PMID- 15780192 TI - ONO-1078 reduces NMDA-induced brain injury and vascular cell adhesion molecule-1 expression in rats. AB - AIM: To determine whether ONO-1078 (pranlukast), a potent cysteinyl leukotriene receptor 1 (CysLT1) antagonist, has an effect on N-methyl-D-aspartate (NMDA) induced brain injury and vascular cell adhesion molecule-1 (VCAM-1) expression in rats. METHODS: Brain injury was induced by direct microinjection of NMDA (0.3 mumol in 1 muL of sterile 0.1 mol/L PBS, pH 7.4) into the cerebral cortex. The lesion volume (area), brain edema and neuron density were assessed by an image analyzer and the expression of VCAM-1 in the cortex was detected by Western blot 24 h after NMDA injection. ONO-1078 (0.03, 0.1, or 0.3 mg/kg) and edaravone (MCI 186, 10 mg/kg), a neuroprotective agent, were ip injected 30 min before and after NMDA injection. RESULTS: NMDA microinjection produced well-defined focal lesions dose- and time-dependently. ONO-1078 (0.1, 0.3 mg/kg) and edaravone (10 mg/kg) decreased the total lesion volume, lesion area and brain edema induced by NMDA. Furthermore, ONO-1078 (0.1, 0.3 mg/kg) significantly inhibited the enhanced expression of VCAM-1 in the injured cortices, but edaravone did not have this effect. CONCLUSION: CysLT1 receptor antagonist ONO-1078 attenuates NMDA-induced brain damage in rats, and this might relate to the attenuation of NMDA receptor dependent neurotoxicity and the inhibition of the upregulation of VCAM-1 expression. PMID- 15780193 TI - Down regulation of cyclooxygenase-2 is involved in delayed neuroprotection by ischemic preconditioning in rats. AB - AIM: To examine whether the prostaglandins (PGs) pathway is involved in triggering delayed neuroprotection by ischemic preconditioning (IPC) and evaluate the effects of IPC on cyclooxygenase-2 (COX-2) expression following focal cerebral ischemia and reperfusion in rats. METHODS: IPC was induced by 10 min of saline infusion into the left internal carotid artery with the right common carotid artery clamped at the same time. Middle cerebral artery occlusion (MCAO) and reperfusion model was produced using intraluminal filament method. RESULTS: IPC 48 h prior to MCAO significantly reduced infarct area as compared with MCAO alone. A nonselective inhibitor of COX indomethacin (3 mg/kg, ip) applied 1 h prior to or 1 h after IPC failed to affect its protective effects. IPC had no direct effect on the cortex COX-2 mRNA and protein expression 72 h later, but decreased the expression of COX-2 mRNA and protein following ischemia and reperfusion insult. CONCLUSION: PGs pathways was not involved in triggering delayed neuroprotection by IPC, and IPC induced down-regulation of COX-2 following focal cerebral ischemia and reperfusion in rats in vivo. PMID- 15780194 TI - Ascorbic acid improves impaired venous and arterial endothelium-dependent dilation in smokers. AB - AIM: To compare the acute effects of ascorbic acid on vasodilation of veins and arteries in vivo. METHODS: Twenty-six healthy non-smokers and 23 healthy moderate smokers were recruited in this study. The dorsal hand vein compliance technique and flow-mediated dilation were used. Dose-response curves to bradykinin and sodium nitroprusside were constructed to test the endothelium-dependent and independent relaxation before and after acute infusion of ascorbic acid. RESULTS: Smokers had an impaired venodilation with bradykinin compared with non-smokers (68.3%+/-13.2% vs 93.7%+/-20.1%, respectively; P<0.05). Ascorbic acid administration in the dorsal hand vein significantly increased the venodilation with bradykinin in smokers (68.3%+/-13.2% vs 89.5%+/-6.3% before and after infusion, respectively; P<0.05) but not in non-smokers (93.7%+/-20.1% vs 86.4%+/ 12.4% before and after infusion, respectively). Similarly, the arterial response in smokers had an impaired endothelium-dependent dilation compared with that in non-smokers (8.8%+/-2.7% vs 15.2%+/-2.3%, respectively; P<0.05) and ascorbic acid restored this response in smokers (8.8%+/-2.7% vs 18.7%+/-6.5% before and after infusion, respectively; P<0.05), but no difference was seen in non-smokers (15.2%+/-2.3% vs 14.0%+/-4.4% before and after infusion, respectively). The endothelium-independent dilation did not differ in both the groups studied. No important hemodynamic change was detected using the Portapress device. CONCLUSION: Smokers had impaired endothelium-dependent vasodilation responsiveness in both arterial and venous systems. Ascorbic acid restores this responsiveness in smokers. PMID- 15780195 TI - Identification and classification of all potential hemolysin encoding genes and their products from Leptospira interrogans serogroup Icterohae-morrhagiae serovar Lai. AB - AIM: To identify and classify all potential hemolysin candidates of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai. METHODS: All of the potential hemolysin encoding genes were characterized in silico. These genes were cloned and expressed in Escherichia coli. The hemolytic activities of the expressed proteins were assayed observing the hemolysis on sheep blood agar plates. Sphingomyelinase activities of the hemolysin candidates were measured by thin-layer chromatography (TLC) and HPLC for sphingomyelin-hydrolysis. Expression and secretion of the hemolysins in L interrogans were studied by reverse transcription polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assays. RESULTS AND CONCLUSION: The hemolytic activities of hemolysin candidates (LA0327, LA0378, LA1027, LA1029, LA1650, LA3050, LA3937, LA4004) from L interrogans strain Lai were confirmed. They were further divided into two groups, sphingomyelinase hemolysins and non-sphingomyelinase hemolysins, based on their ability to hydrolyze sphingomyelin. Most of these hemolysins were actually expressed in living L interrogans and some of them were secreted into the environment. This study establishes an essential and complete basis for further studying the contribution of hemolysins to the pathogenesis of L interrogans. PMID- 15780196 TI - Identification of a novel splice variant of human PD-L1 mRNA encoding an isoform lacking Igv-like domain. AB - AIM: To investigate the expression and regulation of PD-1 ligand 1 (PD-L1) in peripheral blood mononuclear cells (PBMC). METHODS: The cDNA encoding human PD-L1 precursor was cloned from the total RNA extracted from the resting and phorbol dibutyrate plus ionomycin- or phytohemagglutinin-activated PBMC, by reverse transcription polymerase chain reaction (RT-PCR), and independent clones were sequenced and analyzed. The expression and subcellular localization were examined in transiently transfected cells. The PD-L1 gene expression in different PBMC was also analyzed by RT-PCR. RESULTS: A novel human PD-L1 splice variant was identified from the activated PBMC. It was generated by splicing out exon? encoding an immunoglobulin variable domain (Igv)-like domain but retaining all other exons without a frame-shift. Consequently, the putative translated protein contained all other domains including the transmembrane region except for the Igv like domain. Furthermore, the conventional isoform was expressed on the plasma surface whereas the novel isoform showed a pattern of intracellular membrane distribution in transiently transfected K562 cells. In addition, the expression pattern of the PD-L1 splice variant was variable in different individuals and in different cellular status. CONCLUSION: PD-L1 expression may be regulated at the posttranscriptional level through alternative splicing, and modulation of the PD L1 isoform expression may influence the outcome of specific immune responses in the peripheral tissues. PMID- 15780197 TI - Hepatocyte-like cells from directed differentiation of mouse bone marrow cells in vitro. AB - AIM: To design the effective directed differentiation medium to differentiate bone marrow cells into hepatocyte-like cells. METHODS: Bone marrow cells were cultured in the directed differentiation media including fibroblast growth factor 4 (FGF-4) and oncostatin M (OSM). Hepatocyte-like cells from directed differentiation of bone marrow cells were identified through cell morphology, RNA expressions by reverse transcriptase-polymerase chain reaction (RT-PCR), protein expressions by Western blot, and hepatocellular synthesis and metabolism functions by albumin ELISA, Periodic acid-Shiff staining and urea assay. RESULTS: Some epithelial-like cells or polygonal cells appeared and increased in the course of the cell directed differentiation. Hepatocyte nucleur factor-3beta (HNF 3beta, albumin (ALB), cytokeratin 18 (CK18), transthyretin (TTR), glucose-6 phosphate (G-6-Pase), and tyrosine aminotransferase (TAT) mRNA were expressed in the course of the directed differentiation. The directed differentiated cells on d 21 expressed HNF-3? ALB, and CK18 proteins. The directed differentiated cells produced albumin and synthesized urea in a time-dependent manner. They could also synthesize glycogen. CONCLUSION: Our differentiation media, including FGF-4 and OSM, are effective to differentiate bone marrow cells into hepatocyte-like cells, which could be used for hepatocyte resources for bioartificial liver or hepatocyte transplantation. PMID- 15780198 TI - Inducible effects of icariin, icaritin, and desmethylicaritin on directional differentiation of embryonic stem cells into cardiomyocytes in vitro. AB - AIM: To investigate the possible inducible effects of icariin, icaritin, and desmethylicaritin on the directional differentiation of embryonic stem (ES) cells into cardiomyocytes in vitro. METHODS: ES cells were cultivated as embryoid bodies (EBs) in hanging drops with icariin, icaritin, or desmethylicaritin. ES cells treated with retinoic acid and with solvent were used as positive and negative controls, respectively. The cardiomyocytes derived from the ES cells were verified using immunocytochemistry. The expression of cardiac developmental dependent genes was detected using the reverse transcription-polymerase chain reaction (RT-PCR) method. Cell cycle distribution and apoptosis were analyzed using flow cytometry to determine the partly inducible effect mechanisms involved. RESULTS: The total percentage of beating EBs treated with 10(-7) mol/L icariin, icaritin, or desmethylicaritin was 87% (P<0.01), 59% (P<0.01), and 49%, respectively. All the beating cardiomyocytes derived from the ES cells expressed cardiac-specific proteins for a-actinin and troponin T. Among them, 10(-7) mol/L icariin treatment resulted in a significantly advanced and increased mRNA level of a-cardiac major histocompatibility complex (MHC) and myosin light chain 2v (MLC-2v) in EBs in the early cardiac developmental stage. Before shifting to the cardiomyocyte phenotype, icariin could evoke the accumulation of ES cells in G0/G1 and accelerate apoptosis of the cell population (P<0.05). CONCLUSION: Icariin facilitated the directional differentiation of ES cells into cardiomyocytes at a concentration of 10(-7) mol/L. The promoting effect of icariin on cardiac differentiation was related to increasing and accelerating gene expression of a-cardiac MHC and MLC-2v, as well as regulating the cell cycles and inducing apoptosis. PMID- 15780199 TI - Engagement of membrane immunoglobulin enhances Id3 promoter activity in WEHI-231 B lymphoma cells. AB - AIM: We have recently shown that engagement of membrane immunoglobulin (mIg) induced upregulation of inhibitor of differentiation 3 (Id3) mRNA, resulting in growth arrest at G1 phase in WEHI-231 cells. In the present study, we examined whether engagement of mIg will affect promoter activity of the Id3 gene in WEHI 231 cells. METHODS: DNA fragments corresponding to the 5'-flanking region of mId3 gene were amplified by polymerase chain reaction (PCR) using genomic DNA as the template. Three DNA fragments upstream of the transcription start site (+1) of the mId3 gene were subcloned into the luciferase reporter vector PGV-B2. The recombinant constructs were transiently transfected into WEHI-231 cells by an electroporation method. After incubation for 24 h, WEHI-231 cells were stimulated with 10 mg/L anti-IgM or irradiated CD40L-expressing NIH3T3 cells or control NIH3T3 cells for further 24 h, followed by assay for luciferase activity. RESULTS: The luciferase analysis demonstrated that basal promoter activity of the Id3 gene was found in the region between -200 and +54. The Id3 promoter activity was increased 2-fold following stimulation with anti-IgM, but not CD40L, compared with medium alone. CONCLUSION: The mIg-mediated upregulation of Id3 expression is controlled, at least in part, through transcriptional regulation, as assessed by luciferase assay. PMID- 15780200 TI - Effect of 7-hydroxystaurosporine on glioblastoma cell invasion and migration. AB - AIM: To investigate the effect of 7-hydroxystaurosporine (UCN-01), a selective protein kinase C (PKC) inhibitor, on cell growth, migration, and invasion in invasive human glioblastoma U-87MG cells. METHODS: PKC activity was determined based on the PKC-catalyzed transfer of the (32)P-phosphate group from [g (32)P]ATP into a PKC-specific peptide substrate. Cell viability was measured by MTT assay. Cell invasion and migration were evaluated by a Boyden chamber assay and scratch wound assay, respectively. Protein expression was analyzed using Western blot assay. The formation of 3-dimensional cellular aggregates was examined by a cell-cell aggregation assay. RESULTS: UCN-01 treatment resulted in concentration- and time-dependent inhibition of U-87MG cell growth at higher doses (>100 nmol/L), and reduced cell invasion and migration capability at less cytotoxic doses (<100 nmol/L). UCN-01 significantly repressed PKC activity. Consistent with this result, UCN-01 blocked cell invasion stimulated by phorbel 12-myristate-13-acetate (PMA) and ethanol (EtOH), 2 PKC activators. Enforced expression of the tumor suppressor genes BRCA1 and PTEN increased the anti invasion potential of UCN-01. Exposure to UCN-01 caused a dose-dependent increase in cell adhesion molecule E-cadherin. The effect of UCN-01 on the formation of cell-cell aggregation was significantly reduced by the addition of an anti-E cadherin antibody. CONCLUSION: UCN-01 inhibits the invasion and migration of human glioma cells. Accordingly, UCN-01 can have potential clinical applications for the treatment of human glioma metastasis. PMID- 15780201 TI - Predictive model of blood-brain barrier penetration of organic compounds. AB - AIM: To build up a theoretical model of organic compounds for the prediction of the activity of small molecules through the blood-brain barrier (BBB) in drug design. METHODS: A training set of 37 structurally diverse compounds was used to construct quantitative structure-activity relationship (QSAR) models. Intermolecular and intramolecular solute descriptors were calculated using molecular mechanics, molecular dynamics simulations, quantum chemistry and so on. The QSAR models were optimized using multidimensional linear regression fitting and stepwise method. A test set of 8 compounds was evaluated using the models as part of a validation process. RESULTS: Significant QSAR models (R=0.955, s=0.232) of the BBB penetration of organic compounds were constructed. BBB penetration was found to depend upon the polar surface area, the octanol/water partition coefficient, Balaban Index, the strength of a small molecule to combine with the membrane-water complex, and the changeability of the structure of a solute membrane-water complex. CONCLUSION: The QSAR models indicate that the distribution of organic molecules through BBB is not only influenced by organic solutes themselves, but also relates to the properties of the solute-membrane water complex, that is, interactions of the molecule with the phospholipid-rich regions of cellular membranes. PMID- 15780202 TI - Genetic variation of low-density lipoprotein-cholesterol and its clinical implications. PMID- 15780203 TI - Rheumatic fever reappraised. PMID- 15780204 TI - Linkage of the cholesterol 7alpha-hydroxylase gene and low-density lipoprotein cholesterol conditional on apolipoprotein E association: the National Heart, Lung, and Blood Institute Family Heart Study. AB - BACKGROUND: Genetic factors account for approximately 50% of the individual variation in plasma low-density lipoprotein cholesterol (LDL-C) concentrations in the general population. Several candidate genes have been proposed but their relative contributions to the variance in LDL-C are not known, except for apolipoprotein E (apoE). We report here an investigation of the relationship between LDL-C and cholesterol 7alpha-hydroxylase (CYP7), as well as apoE and low density lipoprotein receptor (LDLR), three pivotal genes in LDL metabolism. METHODS: Our study population included more than 200 nuclear families with increased coronary heart disease (CHD) risk from the National Heart, Lung, and Blood Institute (NHLBI) Family Heart Study. Variance-component linkage methods, a measured genotype approach, and a variance-component linkage analysis conditional on a measured genotype association were used. RESULTS: The results showed significant linkage between a genetic determinant of plasma LDL-C concentrations and a polymorphism near CYP7 with its allelic variation accounting for 27% of the total LDL-C variation. There is significant association between plasma LDL-C concentrations and apoE genotypes. Conditional on the apoE association, the total LDL-C variation accounted by allelic variation of a polymorphism near CYP7 was increased significantly. CONCLUSION: Our results suggest the apoE and CYP7 may be two important genes accounting for the genetic variation of plasma LDL-C concentrations in a population with cardiovascular diseases. PMID- 15780205 TI - Detection of herpes simplex virus type 1 in rheumatic valvular tissue. AB - BACKGROUND: Rheumatic heart disease (RHD) is the most important sequela of rheumatic fever (RF): evidence that streptococcal infection is aetiological is prominent, but sometimes contradictory. Acute HSV-1 infection in mouse leads to carditis and valvulitis whereas recurrent infection results in inflammatory granulomatous lesions that resemble Aschoff bodies. Cells containing HSV-1 inclusions or virus infected giant cells appear similar to Anitschkow cells or Aschoff cells respectively. We hypothesized that HSV-1 infection also may be involved in RHD. METHODS: Formalin-fixed, paraffin-embedded valvular tissue samples from 32 patients with RHD were investigated for evidence of HSV-1 infection. HSV-1 antigen was detected by immunohistochemistry, using HSV-1 specific monoclonal and polyclonal antibodies. HSV-1 glycoprotein D gene sequences were amplified by nPCR, using beta-globin gene amplification in the same samples as internal control. Valvular tissue from 5 cases of sudden death and 3 cases died of neisseria meningitis without a history of valvular disease was used for comparison. HSV-1-infected lung tissue was used as positive control. RESULTS: HSV-1 antigens were detected in valvular tissues from 21 of 32 (65.6%) patients. Fifteen of these 21 (46.9% of cases), but no antigen-negative sample, were positive also for HSV DNA. Nucleotide sequence of PCR products was homologous to the targeted region of the HSV-1 glycoprotein D gene. HSV-1 antigen was present also in one case of sudden death but viral DNA was not found in any tissue sample from the comparison group. Results from reagent and positive controls were as anticipated. CONCLUSIONS: This is the first study to show the presence of HSV-1 antigen and genomic DNA in valvular tissues from patients with RHD and provides evidence for an association of HSV-1 infection with some cases of rheumatic valvular disease. PMID- 15780206 TI - The beta-adrenergic blocker carvedilol restores L-type calcium current in a myocardial infarction model of rabbit. AB - BACKGROUND: Carvedilol, an antagonist of alpha1- and beta-adrenergic receptors, has shown efficacy in reducing all-cause death and arrhythmia death for ischemic heart disease and congestive heart failure in several large-scale trials. It has been found to prevent ventricular remodeling, and recently was reported to reverse down-regulation of Na+ channel in a chronic heart failure model. This study was conducted to investigate whether carvedilol could reverse the ion remodeling in a myocardial infarction model of rabbit. METHODS: After the procedure of coronary ligation, animals were randomized to placebo or carvedilol treatment (5 mg/kg). Action potentials, L-type calcium current (Ica L) and the effect of isoproterenol stimulation on Ica L were measured using whole-cell patch method. Evaluation of the expression of calcium channel subunits was carried out by RT-PCR and Western blot. RESULTS: The results indicate that mean peak Ica L densities (pA/pF) at +10 mV was reduced in postinfarction myocytes (5.33 +/- 0.45, n = 25) compared to sham myocytes (6.52 +/- 0.21, n = 20). Treatment of myocardial infarction rabbits with carvedilol could restore it partially (5.91 +/ 0.39, n = 20, P < 0.05). However, steady-state activation parameters were similar in three groups. With stimulation by isoproterenol (1 micromol/L) Ica L increased in all three groups, but the increase was smaller in postinfarction myocytes. mRNA levels of calcium channel subunit CaA1 gene was decreased but CaB2a, CaB2b and CaB3 mRNA levels did not change after MI. Corresponding change in CaA1 protein was also observed. CONCLUSIONS: The results demonstrate that carvedilol restores Ica L density and reverse the downregulation of CaA1 postinfarction. PMID- 15780207 TI - Benefit period using alendronate to increase bone mineral density in women with osteoporosis? AB - BACKGROUND: Alendronate, a nitrogen-containing bisphosphonate is a specific inhibitor of bone resorption and now in the forefront of treatment of osteoporosis. In this study, we reported a significant increase in bone mineral density (BMD) of the spine and the hip in postmenopausal women taking alendronate at 10 mg/d for 1, 2 and 3 years. METHODS: Participants had received daily, oral, 10 mg dose of alendronate for one to three years and placed into one of three groups according to alendronate treatment duration: 41 women received alendronate for 1 year (group I), 46 received alendronate for 2 years (group II), and 30 received alendronate for 3 years (group III). Measurements of bone density had been made by dual energy X-ray absorbtiometry once each year. RESULTS: The differences in L2-L4, L2, L4, femoral neck and trochanter BMD values before and after treatment for first group were significantly different. In second group, significant differences between initial and after treatment were found at the other sites except at the Ward's triangle. In the third group, only a significant increase in the L2-L4, L2, L3, L4, trochanter BMD values between before treatment and at the end of third year was found. Comparisons between groups were performed with Student's t test. ANOVA was used to test the age, menopause age, menopause duration and initial BMD values between the three groups. Calculated P values of less than 0.05 were considered statistically significant. CONCLUSIONS: Alendronate had increased BMD significantly at the spine and hip in postmenopausal women over three years. Increases of BMD in third group were significant during the first and second years. However, continued therapy with alendronate had been required to maintain the gain in BMD over the third year. PMID- 15780208 TI - Effects of extract of Ginkgo biloba with venlafaxine on brain injury in a rat model of depression. AB - BACKGROUND: Recent studies have indicated that chronic stress may give rise to brain damage, which is related to the genesis of depression. The purpose of this study is to investigate the effects of extract of Ginkgo biloba (EGb) and venlafaxine on depression. METHODS: Rats were treated with chronic and comprehensive stress to create a depression model. Immunohistochemistry was used to detect the expression of brain-derived neurotrophic factor (BDNF) in the hippocampal CA3 neurons of rats treated with different drugs. Behavioral changes of these rats were also examined. RESULTS: The expression of BDNF in the hippocampal CA3 neurons of the depression model decreased with a reduction in exploring behavior and a significant increase in fecal production. The expression of neuron nitric-oxide synthase (nNOS) protein also increased in the rats compared to normal controls. The rats treated with EGb and venlafaxine showed an increase in expression of BDNF and exploring behavior compared to untreated rats, but a decrease in nNOS and fecal production. CONCLUSIONS: Rats sustain damage to the brain after being subjected to chronic and comprehensive stress. Our research has indicated that combined EGb with venlafaxine enhances the protection of neurons and decreases damage to the brain, while relieving the side effects of synthetic antidepressants. PMID- 15780209 TI - Expression of c-fos and c-jun proteins in the marginal division of the rat striatum during learning and memory training. AB - BACKGROUND: A new brain region, the marginal division (MrD), was discovered at the caudal margin of the neostriatum. The MrD was shown to be involved in learning and memory in the rat. The aim of this study was to investigate the expression of the immediate-early genes c-fos and c-jun in the MrD of the striatum during learning and memory processes in the rat, immunocytochemical and Western blot methods were used to examine Y-maze trained rats. METHODS: The rats were divided into three groups, namely the training, pseudotraining, and control groups. After Y-maze training, the expression of the immediate-early genes c-fos and c-jun in the MrD of the rats was investigated using immunocytochemical and Western blot methods. RESULTS: After one hour of Y-maze training, the expression of c-jun and c-fos proteins was significantly enhanced in the MrD; the c-jun protein, in particular, was more intensely expressed in this region than in other parts of the striatum. The expression of these two proteins in the training group was significantly higher than in the pseudotraining and control groups. In addition, positive expression was also found in the hippocampus, cingulum cortex, thalamus, and in other areas. Western blot disclosed two immunoreactive bands for the anti-c-fos antibody (47 kD and 54 kD) and two immunoreactive bands for the anti-c-jun antibody (39 kD and 54 kD). CONCLUSIONS: These results indicate that the immediate-early genes c-fos and c-jun participate in signal transduction during the learning and memory processes associated with Y-maze training in rats. PMID- 15780210 TI - The anatomic and electrophysiological characters of the coronary sinus. PMID- 15780211 TI - Effects of hypoxia on promoter of telomerase reverse transcriptase and cell cycle distribution in neonatal rat cardiac myocytes. PMID- 15780212 TI - Coronary artery calcification is associated with atherogenic lipid changes, cardiac dysfunction and morphologic abnormalities in HIV-1 infected black adults. PMID- 15780213 TI - Relationship between IRS-2G1057D variant and type 2 diabetes mellitus in Han population in Liaoning Province, China. PMID- 15780214 TI - The role of oral glucose tolerance test in screening for diabetes mellitus in the elderly of Harbin, China. PMID- 15780215 TI - Detection of nanobacteria in serum, bile and gallbladder mucosa of patients with cholecystolithiasis. PMID- 15780216 TI - Clinical study of 312 cases with matastatic differentiated thyroid cancer treated with large doses of 131I. PMID- 15780217 TI - Clinical and angiographic follow-up study of sirolimus-eluting stent for treatment of in-stent restenosis. PMID- 15780218 TI - Surgical interruption of spinal dural arteriovenous fistulas. PMID- 15780219 TI - Clinical experience with Leksell gamma knife in the treatment of trigeminal schwannomas. PMID- 15780220 TI - The UK breast implant registry-ten years on. PMID- 15780221 TI - Drainage in breast reduction surgery: a prospective randomised intra-patient trail. AB - One hundred and fifty bilateral breast reductions were prospectively randomised intra-operatively to receive a unilateral wound drain. Seven patients required post-operative evacuation of haematoma, three on the drained and four on the undrained sides. There was no statistical difference in haematoma formation between drained and undrained sides (p = 1), or the incidence of wound healing or other complications. We believe that routine wound drainage in breast reduction surgery is an unnecessary intervention. PMID- 15780222 TI - Predicting cup size after reduction mammaplasty. AB - Presently, predicting cup size after reduction mammaplasty is largely based on experience. Regnault and Daniel (1984) proposed a formula for estimating the appropriate weight of reduction to achieve a desired cup size. To validate their method, we studied 75 consecutive patients undergoing bilateral breast reduction mammaplasty. Patients were measured pre-operatively and 6 weeks post-operatively using two techniques to determine bra size. Actual weight of breast tissue excised was compared to the predicted values using the Regnault and Daniel formula for each measuring technique (Methods A and B). A modified formula (Method C) was developed to increase the accuracy. The results obtained using Methods A and B were compared with those of Method C. A paired sample t test showed no significant difference between actual and predicted weight of reduction for using Method A or C. Method B systematically underestimated the weight of tissue to be removed. The mean estimate for Method B was 730 g, a difference of 45 g (CI: -81.7 to -8.9; p = 0.015). For Method A, the mean was 765.5 g, a difference of 9.4 g (CI: -68.1 to 49.3; p = 0.75). This compared to Method C where the mean predicted weight of tissue to be removed was 770.4 g, a difference of -4.5 g (95% CI: -42.4 to 33.4; p = 0.81). The implications of these findings are discussed. PMID- 15780223 TI - Two staged breast reconstruction following prophylactic bilateral subcutaneous mastectomy. AB - The aim of this retrospective study was to evaluate the results in patients who underwent bilateral subcutaneous mastectomy (BSCM) for prophylaxis against invasive breast cancer. All patients were operated on with the same protocol regarding indications and surgical method. Reconstruction was completed in two stages with tissue expanders and permanent round or shaped rough textured gel filled silicone implants. The study includes 52 patients with a mean age of 39.5 years operated on in the period 1991-2000; the period of follow-up ranged between 3 and 12 years with a mean of 7. In this series, not a single case of invasive cancer developed, and the aesthetic results are considered very satisfactory. This data strengthens the case of subcutaneous mastectomy as a valid prophylactic operation. PMID- 15780224 TI - Speech and swallowing following radial forearm flap reconstruction of major soft palate defects. AB - Velopharyngeal function is often compromised by the resection and reconstruction of oropharyngeal and palatal tumours. While free tissue transfer has improved the outcomes of head and neck reconstruction. In general, palatal reconstruction remains a challenge. Velopharyngeal function was analysed in eight patients following microsurgical reconstruction of defects of between 50 and 100% of the soft palate. The radial forearm fasciocutaneous free flap was used in all cases. The outcome of reconstruction was analysed by patient questionnaire and with standardised tests of speech and swallowing function. Velopharyngeal function post-operatively ranged from poor to near normal. Poor function appeared due to the loss of active elevation and contracture of the reconstructed palate producing failure of velopharyngeal closure during swallowing and speech. The results emphasise the limitations of reconstruction of a dynamic structure such as the soft palate with the static fold of skin and soft tissue produced by a fasciocutaneous flap. The relatively poor results obtained suggest that an anatomical approach to soft palate reconstruction is inadequate and reduction of the calibre of the velopharyngeal aperture is required to compensate for the lack of mobility in the reconstructed palate. PMID- 15780225 TI - Secondary repair of cleft lip nose deformity using subcutaneous pedicle flaps from the unaffected side. AB - We used three types of subcutaneous pedicle flaps harvested from the unaffected side of the nostril to repair postoperative nose deformity caused by primary cleft lip surgery. By moving the subcutaneous pedicle flap from a nearby tissue rich area, it was possible to achieve a favourable symmetrical nasal contour for the narrowing of the nostril cavity, depression of the nostril floor and the columella deviation of the affected side. The method, we used here is technically simple. We believe this procedure is a method of choice for repairing small deformities of the nose occurring after primary or secondary cleft lip surgery. PMID- 15780226 TI - Analysis of 109 Japanese children's lip and nose shapes using 3-dimensional digitizer. AB - We assessed lip and nose shapes, which played an important role in performance evaluations, before and after cleft lip and nose surgery. We used a noncontact type semiconductor laser 3-dimensional measurement system on normal Japanese children to obtain 3-dimensional images of noses and lips, which were accurate enough to measure facial shapes. We could rotate these images on the computer, therefore we measured the following points: the distance between the peaks of the Cupid's bow and the width of the labial fissure (frontal view), and the width of the nose and the nasal tip protrusion (basal view). Lip and nose shapes were evaluated for each gender. Additionally, nasolabial angles (NLA) were measured on the lateral views of faces. We classified the morphology of the philtral columns into four types; (1) triangular type, (2) parallel type, (3) concave type, and (4) flat type. We also classified nostril shapes into four types: (1) teardrop type, (2) heart shaped type, (3) round type, and (4) triangular type. We calculated the average of the 3-dimensional coordinate values for each landmark, and created standard facial models of normal Japanese children. Moreover, we invented a new morphological evaluation method before and after cleft lip and nose surgery, using the 3-dimensional data converting and editing software. The method was more feasible to evaluate the assessment cleft lip and nose surgery, by quantifying the surface areas of right and left nostrils and the surface areas of upper and lower vermilions, even now by measuring with the eyes and comparing them. PMID- 15780227 TI - The role of near-infrared angiography in the assessment of post-operative venous congestion in random pattern, pedicled island and free flaps. AB - Indocyanine green near-infrared-video angiography (ICG-NIR-VA) was recently introduced for measuring perfusion of skin flaps. The prognostic value of this method with regards to post-transfer manipulations of the flap, and subsequently, flap survival is not adequately documented in the literature. In this paper, we report our experience with the ICG-NIR-VA in the intraoperative evaluation and post-operative follow-up of nine flaps (2 large random pattern, 4 pedicled island and 3 free flaps) used in various reconstructive procedures. Two flaps (1 random pattern and 1 free flap) showed delayed intraoperative uptake in ICG-NIR-VA. In the post-operative phase, (days 1-3) six flaps (1 random pattern, 2 axial pattern and 3 free flaps) showed a delay both in the ICG-NIR-VA uptake, as well as clearance. However, a clinical correlate was observed only in 2 of the 6 flaps demonstrating this delay: general and partial venous congestion was seen in a distally based interosseous posterior flap and a free lateral arm flap respectively. Leeches were implemented only based on the clinical signs. In one distally based perforator flap, the decision on perfusion augmentation via microanastomosis was based on the intraoperative ICG-NIR-VA finding. All flaps showed uneventful healing. Based on our observations, the question arises, not whether the ICG-NIR-VA is sensitive for the prognosis of venous congestion, upon which the flap manipulation strategy might rest-but whether it is too sensitive. Further, prospective studies are necessary. PMID- 15780228 TI - Vascular endothelial growth factor gene therapy with intramuscular injections of plasmid DNA enhances the survival of random pattern flaps in a rat model. AB - The objective of this study was to determine the effects of the naked plasmid DNA encoding vascular endothelial growth factor (VEGF) on the survival of random flaps on rats. Thirty Sprague-Dawley rats whose random flaps were elevated on the back were randomised into three groups of 10 animals each. In the experimental group, the naked plasmid DNA encoding VEGF was injected directly into the panniculus carnosus of the flap. In the two control groups, either control plasmid DNA or physiologic saline was injected. After 7 days, the flaps were evaluated with the following devices: RT-PCR for the expression of VEGF gene, immunohistochemistry for the expression of VEGF protein, histology for vascular density, single photon emission computerised tomography for RBC in the flap, and image analysis for flap survival area. Notably increased expressions of VEGF mRNA and VEGF protein were found in the treatment group. Vascular density was markedly more increased in the treatment group than those in the two control groups (P < 0.01). Compared with the two control groups, the flap treated with VEGF plasmid DNA showed a more significantly enhanced tissue viability: 87 +/- 5 versus 47 +/- 6% for the control plasmid DNA group and 46 +/- 5% for the saline group (P < 0.01). Our results indicated that the VEGF gene therapy was able to enhance the survival of random pattern flaps by inducing angiogenesis. PMID- 15780229 TI - GLUT-1: an extra diagnostic tool to differentiate between haemangiomas and vascular malformations. AB - The differential diagnosis between juvenile haemangiomas, vascular malformations, pyogenic granulomas and normally proliferative endothelium (granulation tissue) on the basis of histology alone is sometimes difficult. This is important because haemangiomas, are self-limiting and vascular malformations are not. We report our experience of using the immunohistochemical marker GLUT-1 to distinguish haemangiomas from vascular malformations following the initial report by North and Colleagues (1998). We studied a total of 50 specimens from patients with vascular anomalies, and found that GLUT-1 reactivity was positive in 18 out of 19 juvenile haemangiomas, negative in two out of two noninvoluting congenital haemangiomas (NICH) and negative in 29 out of 29 vascular malformations, that included capillary malformations, lymphatic malformations, venous malformations and arteriovenous malformations (95% sensitivity, 100% specificity). Pyogenic granulomas (n = 4) and granulation tissue samples (n = 4) were used as negative controls. Placenta tissue was used as positive control. GLUT-1 accurately distinguishes haemangiomas from vascular malformations, and as a result from this work, we use this technique in routine histopathological differentiation of vascular anomalies. PMID- 15780230 TI - Ectropion following excision of lower eyelid tumours and full thickness skin graft repair. AB - A retrospective review was performed to define the incidence of post operative lower eyelid ectropion after excision and full thickness grafting of lower eyelid tumours. In a 7-year period (1995-2001) 106 consecutive patients were identified who had undergone such surgery with mean follow-up period of 7.4 months (median 5 months, range 1-41.5 months). The majority of lesions were basal cell carcinomas 89/106 (84%). A total of 15/106 (14.2%) patients developed ectropion. The length of follow-up for patients with ectropion was on average 11.7 months (median 10 months, range 2-34 months). Ten patients underwent corrective surgery with five of these showing cosmetic and symptomatic improvement. Of five patients treated conservatively, three showed spontaneous improvement. The incidence of lower eyelid ectropion after lower eyelid skin grafting is defined and this does not seem to have been reported before for a consecutive series of patients. We offer a classification for eyelid zones with regard to ectropion and anatomical and pathological factors that may be important for ectropion formation are discussed. Finally we suggest a new classification of ectropion based on cosmetic and functional consequences. PMID- 15780231 TI - Reconstruction of seventeen full-thickness defects of the eyelids with twenty-two Hubner tarsomarginal grafts. AB - In 1976, Hubner described a new technique to repair full-thickness defects of the eyelids. In 1993, we decided to adopt this simple and easily reproducible technique that guarantees restoration of the normal aspect of the margin. Subsequently, 17 eyelid reconstructions were performed on 13 patients, requiring the harvesting of 22 tarsomarginal grafts. No cosmetic or functional sequelae in the donor eyelids were observed. One or several functional complications were present in seven out of 17 reconstructed eyelids, including two cases of epiphora, one case of lagophtalmos and two cases of lid notch. Loss of the eyelids was observed in three cases out of six upper eyelid reconstructions and in nine cases out of 11 lower eyelid reconstructions. In all cases, the margin integrity was otherwise preserved. Very few simple techniques achieve that level of quality in the eyelid reconstruction process. PMID- 15780232 TI - The Cook-Swartz venous Doppler probe for the post-operative monitoring of free tissue transfers in the United Kingdom: a preliminary report. AB - Accurate assessment of the perfusion of free tissue transfers has always been a challenge for surgeons undertaking microvascular reconstructive procedures. The complexities of flap microcirculation are often difficult to assess despite all the subjective and objective examination techniques available today, particularly when the free tissue transfer is buried, and not visible for monitoring. The Cook Swartz venous Doppler system is a technique for monitoring venous flow in free tissue transfer consisting of an implantable, removable, 20 MHz ultrasonic probe around the venous pedicle and a battery operated portable monitor. We perceive it as a quick and easy to use system, which in our study was well received by both medical and nursing staff. It can be used in conjunction with other monitoring techniques and we found it of value following revascularisation, during inset and in post-operative monitoring of free flaps particularly when operating outside our base hospital. We believe our initial experiences, on 24 patients, with the device, supports the use of a Cook-Swartz probe as an adjunct to traditional clinical monitoring techniques. We have had no technical difficulties with its application, use and removal, so far and we plan to continue with its use when it becomes available outside of a clinical trial. PMID- 15780233 TI - The compound thoracodorsal perforator flap in the treatment of combined soft tissue defects of sole and dorsum of the foot. AB - Nine cases of massive soft-tissue loss of the foot were reconstructed by means of a compound (chimera) thoracodorsal artery perforator (TAP) flap, which reconstituted the different functional units (dorsum, heel, instep, weight bearing surface). In each case, the flap consisted of a skin component isolated on its perforator in combination with a portion of latissimus dorsi muscle and/or serratus fascia, all pedicled on the thoracodorsal vessels. The pedicle length allows up to 4-6 cm of independent mobility of the skin island. The mobility of the various flap components allows the various functional units of the foot to be reconstructed without relying on multiple flaps or anastomoses. The pedicle length was sufficient to be able to perform the anastomosis out of the zone of injury. In some cases the skin island was harvested along with intercostal nerve branches, this gave us the potential to develop a sensate flap. The indications and advantages of this reconstructive method are discussed. PMID- 15780234 TI - Effects of an informational CD-ROM on anxiety and knowledge before aesthetic surgery: a randomised trial. AB - This trial aimed to assess the value of visual information by CD ROM before aesthetic breast reduction or abdominoplasty. All included patients were given an information leaflet about the procedure and completed a Spielberger state-trait anxiety inventory (STAI form Y-1) questionnaire to assess situational anxiety. The patient was then randomly assigned to watch or not to watch an informational CD ROM. The day before surgery the patient completed a second anxiety questionnaire and a knowledge questionnaire. Eighty patients were included, 40 were assigned the CD ROM and 40 no CD ROM. Patients who watched the CD ROM were significantly less anxious than those who did not (Mean STAI 45 [38.2-46.3] vs 55 [49.9-63.8]). Furthermore they also scored higher in the knowledge questionnaire as far as the purpose and the procedural details of the surgery were concerned. However, no statistical difference of knowledge regarding the potential complications of the procedure was found. PMID- 15780235 TI - A scanning electron microscopical study of the two sides of polypropylene mesh (Marlex) and PTFE (Gore Tex) mesh 2 years after complete abdominal wall reconstruction. A study of 15 cases. AB - The use of biomaterials for the repair of abdominal wall defect is becoming common and safe. It has been 20 years since the senior author developed a method to reconstruct the very large transfixing abdominal wall defect with a combination of two biomaterials (Gore Tex) PTFE as a neo peritoneum and polypropylene superficial to this in order to give rigidity to the abdominal wall) and a superficial flap. An observation at the electron microscopy level of the two sides of the implants' surfaces was performed. At the time of a late abdominal wall surgical revision on 15 patients, the prosthesis fragments have been analyzed at the electron microscopy level. The aim of our study was to analyze the late evolution of the different sides of these prostheses. Our results showed, for the first time in vivo, that there is an impressive stability of the deep side of PTFE ultra structure after implantation, a significant difference of the two sides of PTFE at the ultrastructural level and the creation of an intermediate tissue between the two meshes. In contrast, the polypropylene invariably gave rise to adhesions and colonisation by the surrounding tissues. Findings confirmed that the structure and porosity of a biomaterial play a key role in the appearance of adhesions and their consistency. PMID- 15780236 TI - An alternative approach for correction of constricted ears of moderate severity. AB - Moderately severe constricted ears are characterised by lidding, moderate reduction of the vertical height of the upper ear (reduced scapha, absent superior crus) and prominence with absent antihelix. Techniques described for the correction of moderate constricted ear deformities included an attempt to expand the upper part of the ear by various cartilage transection and/or flap techniques. In the current paper, an alternative approach is presented and is based on the recreation of an antihelix (with Mustarde sutures) and excision of the lidding without performing cartilage expansion procedures. The technique was used in four patients with satisfactory results despite the persistence of the deficient scapha and triangular fossa. No recurrence of the cupping deformity was observed (follow up ranged from 6 months to 4 years with a mean of 2 years) and this was attributed to the use of a 'mastoid hitch' as an adjunctive technique to prevent recurrence. PMID- 15780237 TI - What should be the treatment modality in giant cutaneous lipomas? Review of the literature and report of 4 cases. AB - We present three females (20, 39 and 58 years old) and one male patient (47 years old), admitted to our clinic with 3-20 years history of giant lipomas on periscapular (n = 2), posterior cervical (n = 1) and abdominal areas (n = 1), respectively. The sizes of the tumours ranged from 15 x 14 cm(2) to 35 x 25 cm(2) weighing 1900-12 350 g. In all cases, the diagnosis of the lesions was by physical examination and confirmed with computed tomography, magnetic resonance imaging and ultrasonography. Incisional biopsy or aspiration cytology were used to exclude liposarcoma. Three lesions with surrounding capsule formation and lipofibromatous changes underwent open surgery. In one case, as the capsule formation was not evident, suction assisted lipectomy was attempted. However, no aspirate could be evacuated possibly due to the fibrotic nature of the lesion and open surgery was performed. The follow-up period was 14 months to 5 years and no recurrence was noted. PMID- 15780238 TI - The congenital cervical midline cleft. Case report and review of literature. AB - Congenital midline cervical clefts (CMCC) are rare. There have been approximately 50 reports in the international literature with the majority found in Caucasian females. They consist of a skin tag, mucosal surface and a caudal sinus in the ventral midline of the neck. The embryological cause is disputed, as is the best course of treatment. A case of a 4-month-old Indian boy with a congenital midline cervical cleft is reported. The appropriate investigations are described as well as a new method of correcting the lesion. In addition, the most likely aetiological basis for the condition are discussed. PMID- 15780239 TI - Rehabilitation after the replantation on a 2-year-old girl with both amputated legs. AB - We had an opportunity to perform replantation of both legs on a 2-year-old girl, and our decision to perform replantation rather than amputation surgery was carefully made taking her age, degree of crushing injury, ischaemic time and level of the amputation into consideration. Painstakingly designed rehabilitation treatments were continuously performed on this girl from the early stage after the operation, and the treatments were comprised of four parts; that is, flexion and extension exercise for the ankle in order to prevent it from stiffness or contracture, functional electrical stimulation (FES) in order to prevent muscular atrophy on the lower extremities, muscle strengthening exercise for the lower extremities, and electrical stimulation to regenerate the damaged nerves and to prevent muscular atrophy from occurring. For an objective assessment of the postoperative conditions, total active motion angles of the ankle joint were measured, and also EMG and NCV were conducted at the end of the first month as well as at the end of the 6th month. Total active motion angles of the ankle joint were increased progressively as time went on, from 15 to 60 degrees on the right and from 10 to 45 degrees on the left. NCV did not show any sensation or response from motor nerves, or amplitude decreased considerably 1 month after the operation; however, at the end of the 6th month conditions improved a great deal with both amplitude and latency. And most muscles that did not show any signals on EMG or showed less than normal at the end of the first month after the operation eventually recovered at the end of the 6th month. The patient had no particular difficulties in walking after 6 months or rather she started running in small steps showing her legs functioning superbly. An infant with both of lower extremities amputated is quite a rare case. We believe that the replantation surgery was successful due to the fact that carefully selected preoperative factors were taken into consideration and well designed postoperative rehabilitation program consisted of four parts was carried out continuously. PMID- 15780240 TI - Could your case of necrotising fascitis be Pyoderma gangrenosum? AB - Necrotising fascitis is a fulminant infective process which can result in significant morbidity and can indeed be life threatening. It needs to be recognised early and managed aggressively with extensive surgical debridement followed by appropriate antibiotics. On the other hand, Pyoderma gangrenosum is a disease process that needs to be managed non-surgically. We present one such case, the diagnosis of which had treaded along the thin line between the two. PMID- 15780241 TI - Carney syndrome and the plastic surgeon: presentation, recognition, and evaluation. AB - Carney syndrome is a rare clinical entity usually treated by plastic surgeons. We present a patient with Carney syndrome and review the literature emphasising the plastic and reconstructive surgical aspects of the syndrome. PMID- 15780242 TI - The use of Integra in an upper extremity avulsion injury. AB - We present the case of a 22-year-old man who suffered an avulsion injury of the left upper extremity including the elbow region in an industrial accident. After debridement of the skin flap, the defect was primarily closed with Integra. On day 22 split thickness skin graft was performed. Functional and aesthetic outcome and skin quality are excellent. PMID- 15780243 TI - The palmaris longus tendon-rationality of the finger manoeuvres. PMID- 15780244 TI - 'Thumb protection splints' for reposition of the acute temporo-mandibular dislocation. PMID- 15780246 TI - Experimental approaches to hyaluronan structure. AB - A review of the literature describing experimental studies on hyaluronan (HA) is presented. Methods sensitive to the hydrodynamic properties of HA, analyzed in neutral aqueous solution containing NaCl at physiological concentration, can be shown to fit the expected behavior of a high molecular weight linear semi flexible polymer. The significant nonideality of HA solutions can be predicted by a simple treatment for hydrodynamic interactions between polymer chains. Nuclear magnetic resonance and circular dichroism studies of HA are also in agreement with a model incorporating dynamically formed and broken hydrogen bonds, contributing to the semi-flexibility of the polymer chain, and segmental motions on the nanosecond time scale. HA shows the capability for self-association in the formation of a viscoelastic putty state at pH 2.5 in the presence of salt, and a gel state at pH 2.5 in mixed organic/aqueous solution containing salt. Ordered and associated structures have also been observed for HA on the surfaces, especially in the presence of surface-structured water. These phenomena can be understood in terms of counterion-mediated polyelectrolyte interactions. The possibility that hyaluronan exists in vivo in environments that induce ordered structures and assemblies is discussed. PMID- 15780247 TI - Challenges for the modern analytical ultracentrifuge analysis of polysaccharides. AB - This article reviews some of the recent advances in analytical ultracentrifugation and how these advances have impacted--and can impact--on our understanding of the size, shape through conformation modelling, interactions and charge properties of polysaccharides in solution, particularly when used in combination with other solution techniques and also imaging techniques. Specifically we look at (1) polysaccharide polydispersity and simple shape analysis by sedimentation velocity, and in particular using new approaches such as SEDFIT analysis; (2) polysaccharide molecular-weight analysis by sedimentation equilibrium and MSTAR analysis and how this complements analysis of size exclusion chromatography coupled to multi-angle laser light scattering; (3) polysaccharide conformation analysis using traditional procedures such as the Wales-van Holde ratio, power law or 'scaling' relations, more specialised treatments for rigid cylindrical structures, semi-flexible chains and worm-like coils and complications through draining effects; (4) Analysis of polysaccharide interactions and in particular complex formation phenomena, focusing on interesting applications in the areas of mucoadhesion and sedimentation fingerprinting; and (5) the possibilities for macromolecular charge and charge screening measurement. PMID- 15780248 TI - Determining the crystal structure of cellulose III(I) by modeling. AB - Recently, a one-chain monoclinic unit cell for cellulose III(I) having P2(1) symmetry and a single glucose in the asymmetric unit was proposed, based on high resolution diffraction patterns. The new work challenged a two-chain structure that was published 25 years earlier, although it did not provide new three dimensional coordinates. Our goals were to solve the structure by modeling, find whether modeling would reject the previously determined two-chain unit cell, and compare the model with the anticipated experimental structure. Combinations of three rotamers of the O-2, O-3, and O-6 hydroxyl groups produced 27 'up' and 27 'down' starting structures. Clusters ('minicrystals') of 13 cellotetraose chains terminated by methyl groups for each of the 54 starting structures were optimized with MM3(96). Hydroxyl groups on 16 of these 54 structures reoriented to give very similar hydrogen-bonding schemes in the interiors, along with the lowest energies. Hydrogen bonds included the usual intramolecular O-3H...O-5' linkage, with O-6' also accepting from O-3H. Interchain hydrogen bonds form an infinite, cooperative O-6H...O-2H...O-6 network. Direct comparison of total minicrystal energies for the one- and two-chain unit cell was inappropriate because the two chain cell's alternate chains are shifted 0.9 A along the z-axis. To get comparable energy values, models were built with both cellotetraose and cellohexaose chains. The differences in their energies represent the energies for the central layers of cellobiose units. The one-chain cell models had much lower energy. The eight best 'up' one-chain models agree reasonably well with the structure newly determined by experiment. PMID- 15780249 TI - Polysaccharide structures from powder diffraction data: molecular models of arabinan. AB - X-ray intensity data from a polycrystalline sample of debranched arabinan, [-->5) alpha-L-Ara(f)-(1-->](n), have been obtained using a powder diffractometer in order to determine its three-dimensional structure. The observed peaks index on a monoclinic cell with a=5.444(7), b=6.395(10), c=8.680(5) A, and gamma=99.6(3) degrees , V=298 A3. One 2-fold helix along the c-axis can be accommodated in the unit cell. Molecular and packing models have been analyzed using the seven C-2' endo/C-3'-endo allomorphs originally proposed by Radha and Chandrasekaran [Carbohydr. Res. 1997, 298, 105]. The generated powder pattern matches closely with the observed diffraction only for one C-2'-endo model. In this structure, the three main chain conformation angles are in the trans domains, there are no intra-chain hydrogen bonds, and the packing arrangement is stabilized by inter chain O-3-H...O-2 bonds. PMID- 15780250 TI - The two-conformer hypothesis: 2,3,4,6-tetra-O-methyl-mannopyranosyl and glucopyranosyl oxacarbenium ions. AB - Computational chemistry can give information about the probable conformations of reactive intermediates that are difficult to determine experimentally. Based on density functional theory (DFT) calculations of tetra-O-methyl-D-mannopyranosyl and -glucopyranosyl oxacarbenium ions, two families of conformations, which we call B0 and B1, were found. For the manno configuration, a 4H3 and 3E almost isoenergetic pair were found, whereas for the gluco-configuration a 4H3 and 5S1 pair favouring 4H3 were calculated. These results corroborate earlier results and suggest that this two or more conformer hypothesis is general. Nucleophilic attack on these pairs of cations was modelled with methanol and led to four cases to consider namely alpha- or beta-attack on B0 or B1. The resulting complexes (G0, G1 and F0, F1) demonstrate facial selectivity. The relative energies of these complexes are dominated by intramolecular hydrogen bonding and the conformational consequences to the pyranose ring of changes in the C-5-O-5-C-1-C 2 torsion angle. Constrained variation of the nucleophilic oxygen (methanol) to C 1 distance shows that these ion dipole complexes are the only minima with this constraint. PMID- 15780251 TI - The external-anomeric torsional effect. AB - The rotational barrier for a methyl group at the end of an anomeric system is sometimes lower than we might have anticipated. Thus, in the trans-trans conformation of dimethoxymethane, the barrier to methyl rotation is calculated (B3LYP/6-311++G(2d,2p)) to be 2.22 kcal/mol, just slightly smaller than the corresponding barrier to rotation of the methyl group in methyl propyl ether of 2.32 kcal/mol. However, if the methyl being rotated in dimethoxymethane is placed into a gauche conformation, that rotational barrier is reduced to 1.52 kcal/mol. This substantial (0.80 kcal/mol relative to methyl propyl ether) reduction in barrier height in the latter case is attributed mainly to the change in the bond order of the C-O bond to which the methyl is attached, as a function of conformation, which in turn is a result of the anomeric effect. We have called this barrier lowering the external-anomeric torsional effect. This effect is apparently widespread in carbohydrates, and it results in the changing of conformational energies by up to about 2 kcal/mol. If polysaccharide potential surfaces are to be accurately mapped by molecular mechanics, this effect clearly needs to be accounted for. PMID- 15780252 TI - The utility of residual dipolar couplings in detecting motion in carbohydrates: application to sucrose. AB - The solution structure and dynamics of sucrose are examined using a combination of NMR residual dipolar coupling and molecular mechanics force fields. It is found that the alignment tensors of the individual rings are different, and that fitting 35 measured residual dipolar couplings to structures with specific phi, psi values indicates the presence of three major conformations: phi, psi=(120 degrees ,270 degrees), (45 degrees, 300 degrees) and (90 degrees ,180 degrees). Furthermore, fitting two structures simultaneously to the 35 residual dipolar couplings results in a substantial improvement in the fits. The existence of multiple conformations having similar stabilities is a strong indication of motion, due to the interconversion among these states. Results from four molecular mechanics force fields are in general agreement with the experimental results. However, there are major disagreements between force fields. Because fits of residual dipolar couplings to structures are dependent on the force field used to calculate the structures, multiple force fields were used to interpret NMR data. It is demonstrated that the pucker of the fructofuranosyl ring affects the calculated potential energy surface, and the fit to the residual dipolar couplings data. Previously published 13C nuclear relaxation results suggesting that sucrose is rigid are not inconsistent with the present results when motional timescales are considered. PMID- 15780253 TI - Ramachandran free-energy surfaces for disaccharides: trehalose, a case study. AB - We present calculated potential of mean force surfaces for rotation about phi, psi dihedral angles of the alpha(1<-->1)alpha-glycosidic linkage in the disaccharide trehalose (alpha-D-Glc-(1<-->1)-alpha-D-Glc) in both vacuum and aqueous solution. The effects of aqueous solvation upon the alpha(1<-->1)alpha glycosidic linkage are investigated through comparison of the vacuum and aqueous solution free-energy surfaces. These surfaces reveal that trehalose is restricted to a single minimum-energy conformation in both vacuum and solution. The exceptional rigidity of this disaccharide in solution may provide a molecular rationale for the antidesiccant properties of trehalose glasses. PMID- 15780254 TI - Influence of homologous disaccharides on the hydrogen-bond network of water: complementary Raman scattering experiments and molecular dynamics simulations. AB - A comparative investigation of trehalose, sucrose, and maltose in water solution has been performed using Raman scattering experiments and Molecular Dynamics simulations. From the analysis of the O-H stretching region in the [2500,4000] cm(-1) Raman spectral range, which includes for the first time the contribution of 'free' water, and the statistical distribution of water HB probabilities from MD simulations, this study confirms the privileged interaction of trehalose with water above a peculiar threshold weight concentration of about 30%. The role of the hydration number of sugars--found higher for trehalose--on the destructuring effect of the water hydrogen bond network is also addressed. The analysis of the water O-H-O bending spectral range [1500,1800] cm(-1) reveals a change of the homogeneity of water molecules influenced by sugars, but the three investigated sugars are found to behave similarly. PMID- 15780255 TI - Water T2 relaxation in sugar solutions. AB - 1H spin-spin relaxation times of water were measured with the CPMG sequence in dilute aqueous solutions of glucitol, mannitol, glycerol, glycol, the methyl D pyranosides of alpha-glucose, beta-glucose, alpha-galactose, beta-galactose, alpha-xylose, beta-xylose, beta-arabinose and sucrose, alpha,alpha-trehalose, beta-maltose, maltotriose and maltoheptaose. The relaxation-time dispersion was measured by varying the CPMG pulse spacing, tau. These data were interpreted by means of the Carver-Richards model in which exchange between water protons and labile solute hydroxyl protons provides a significant contribution to the relaxation. From the dependences on temperature and tau, parameters characteristic of the pool of hydroxyls belonging to a given solute were extracted by nonlinear regression, including: the fraction of exchangeable protons, P, the chemical-shift difference between water protons and hydroxyl protons, deltaomega, the intrinsic spin-spin relaxation time, T2, and the chemical exchange rate, k. These solute-specific parameters are related, respectively, to the concentration, identity, mobility and exchange life-time of the hydroxyl site. At 298 K, values of deltaomega, T2 and k were found to be of the order of 1 ppm, 100 ms and 1000 s(-1), respectively. Effects of molecular size, conformation and solute concentration were investigated. The exchange mechanism was characterised by Eyring activation enthalpies and entropies with values in the ranges 50-70 kJ mol(-1) and -10 to 60 J K(-1)mol(-1), respectively. PMID- 15780256 TI - Towards understanding the interaction between oligosaccharides and water molecules. AB - Complex carbohydrates are implicated in many important biological processes, and have a strong interaction with water. This close interplay with molecular water through multiple hydroxyls may be an integral part of their emergent structure and dynamics, as selected during evolution. Using molecular dynamics simulations with explicit water the interactions at the linkages within a variety of oligosaccharides are investigated and contrasted, in order to establish correlations between linkage orientation, sugar epimerization, and water interaction. In particular, interactions at alpha linkages, and between mannose and glucose residues, that are common in oligosaccharides are considered. Sugars joined by alpha linkages at the 2-, 3-, and 6-position were found to interact via a combination of weak hydrogen-bonds and water-bridges, which is dependent on the epimerization state of the sugars. Due to their three-dimensional structure, they are also likely to interact with noncontiguous sugar residues in an oligosaccharide, which can lead to ordered structures through the exclusion of water. On the other hand, beta linkages (to 3- and 4-position) maintain strong hydrogen-bonds, have a limited ability to be involved in water-bridges, and predominantly interact with the directly attached sugars. Therefore, sequences of alpha-linked sugars form compact, branched structures that have conformational flexibility, and beta linkages form extended, relatively rigid structures, suitable for structural molecules, and at the termini of protein bound oligosaccharides. These results provide further tentative ties between chemical structure, water interactions, and the emergent form and function of specific sugars and linkages in oligosaccharides. PMID- 15780257 TI - Proton fluctuations and water diffusion in dextran chemical hydrogels studied by incoherent elastic and quasielastic neutron scattering. AB - Proton fluctuations reporting local motions of the glycosidic linkages of chemically crosslinked dextran hydrogels with well defined pore-size distributions are studied by static and dynamic neutron-scattering approaches. The dependence of the dynamic behaviour of water on the pore sizes is also discussed. PMID- 15780259 TI - The disordered conformation of kappa-carrageenan in solution as determined by NMR experiments and molecular modeling. AB - The conformation of kappa-carrageenan in solution was studied combining 1H and 13C NMR with molecular mechanics. The experimental conditions were chosen to characterize the disordered conformation of the polymer. Particular attention has been given to explore a wide range of experimental conditions as to the dependence on solvent (water and Me2SO), polymer concentration, temperature, pH, presence of a denaturing agent (guanidinium chloride), and of ions otherwise able to induce conformational order of the carrageenan chains, either in solution (I-) or in the gel state (Rb+). Two-dimensional NOE experiments were analyzed to obtain information on internuclear distances, and molecular mechanics provided the range of energetically accessible conformations. Two inter-residue topological constraints were clearly identified: their combination is rather restricting for the chain and suggests that the disordered conformation of kappa carrageenan is characterized by an intrinsic stiffness with high values of persistent length and characteristic ratio. They also rule out any postulated interchain hydrogen bonds. In contrast, experiments on the temperature dependence of the chemical shift in Me2SO reveal the existence of two inter-residue intramolecular H-bonds which might contribute positively to the rigidity of the polymer chain. The overall picture emerging from the present results is that of a locally elongated 'loose single helix'. PMID- 15780258 TI - Hyaluronan conformations on surfaces: effect of surface charge and hydrophobicity. AB - Extended, relaxed, condensed, and interacting forms of the polysaccharide hyaluronan have been observed by atomic force microscopy (AFM). The types of images obtained depend on the properties of the surfaces used. We have investigated several different surface conditions for HA imaging, including unmodified mica, mica chemically modified with two different kinds of amino terminated silanes (3-aminopropyltriethoxysilane and N-trimethoxysilylpropyl N,N,N-trimethylammonium chloride), and highly oriented pyrolytic graphite. We found the degree of HA molecular extension or condensation to be variable, and the number of bound chains per unit area was low, for all of the mica-based surfaces. HA was more easily imaged on graphite, a hydrophobic surface. Chains were frequently observed in high degrees of extension, maintained by favorable interaction with the surface after molecular combing. This observation suggests that the HA macromolecule interacts with graphite through hydrophobic patches along its surface. AFM studies of HA behavior on differing surfaces under well controlled environmental conditions provides useful insight into the variety of conformations and interactions likely to be found under differing physiological conditions. PMID- 15780260 TI - Hyaluronan chain conformation and dynamics. AB - An overview of the present state of research in the field of hyaluronan chain conformational aspects is presented. The relationship between structure and dynamics are illustrated for a series of hyaluronan oligomers. Conformational characteristics of hyaluronan chains are discussed, together with the dynamic chain patterns, evaluated by using a theoretical approach to diffusive polymer dynamics. The dependence of correlation times and NMR relaxation parameters from the chain dimension are investigated. Topological features and dimensional properties are related to the structural determinants by using classical computational methods of molecular mechanics and Monte Carlo simulation. PMID- 15780261 TI - Probing macromolecular architectures of nanosized cyclic structures of (1-->3) beta-D-glucans by AFM and SEC-MALLS. AB - Comb-like branched (1-->3)-beta-D-glucans dissolve in water as stiff triple helical structures. Dissociation followed by re-association leads to the formation of a blend of various macromolecular topologies, where the cyclic species make up a significant fraction. In this study, the molecular properties of these nanosized cyclic structures of (1-->3)-beta-D-glucans were probed using a combination of AFM and SEC-MALLS. The cyclic structures were obtained by subjecting linear triple-helical molecules of (1-->3)-beta-D-glucans to a denaturation-renaturation cycle, and the fraction of cyclic structures in the renatured sample was determined by AFM. Samples containing different known fractions of linear and circular molecules were studied by SEC with online multi angle laser-light scattering and viscometric detectors. The molecular weight and the radius of gyration of the molecules eluting from the SEC column, as well as the concentration and the intrinsic viscosity, were determined simultaneously. By extrapolating the results to a situation of only circular species, the results allowed to determine the linear mass per unit length (M(L)) of not only the linear but also the circular morphologies of the (1-->3)-beta-D-glucans. The values obtained were M(L)=2140+/-180 g mol(-1)nm(-1) for the circular species and 2045+/-80 g mol(-1)nm(-1) for the linear species. This is the first direct determination of the M(L) parameter of the circular topology, and the results indicate that the reassociation of the individual chains yield a triplex structure also for the circular morphology, similar to the initial triple helix. PMID- 15780262 TI - The complex of xylan and iodine: the induction and detection of nanoscale order. AB - The complex of xylan and iodine and its formation in a solution of xylan, CaCl2, and I2+KI was investigated by UV/Vis, second-derivative UV/Vis, and Raman spectroscopy. The complex forms only at very high concentrations of CaCl2, suggesting that when the water available in the solution is not sufficient to fully hydrate the calcium cation the chelation with the hydroxyl groups of the xylan can occur. The electronic spectra indicate that iodine is present in the form of three linear polyiodides I9(3-), I11(3-), and I13(3-) structures, which the Raman spectra show to be linear aggregates of the I3- and I5- substructures. Iodide concentration has a significant influence on the relative population of I9(3-), I11(3-), and I13(3-), as well as I3- and I5-, which lead to changes in both the UV/Vis absorption maxima shifts and changes in the Raman spectra. The key difference between this system of complexes with the linear polyiodide aggregates and that of amylose is that the longest aggregate observed with the amylose system, the I15(3-) polyanion, is not observed with the xylans. This indicates that the ordered arrays in the xylan-iodine complex do not exceed 4 nm in length. It is not possible to conclude at this time whether the ordered segment of the xylan molecule is linear or helical. If it is linear the length of the longest ordered arrays would be eight xylose residues. The number would exceed eight if the xylan molecule were helically wound. PMID- 15780263 TI - Effect of the carbohydrate side-chain on the conformation of a glycoconjugate polystyrene in aqueous solution. AB - An oligomaltose-carrying polystyrene "glycoconjugate polystyrene" was synthesized by the homopolymerization of 4-vinylbenzylamine oligomaltonic amides, derived from maltose, maltotriose, maltopentaose, and maltoheptaose. The resultant amphiphilic glycoconjugate polystyrenes were dissolved in 0.1 M aqueous urea, and their structures characterized by small-angle X-ray scattering and molecular modeling. "Glycoconjugate polystyrene" was found to behave as a "molecular bottle brush", composed of a large pseudo-helical polystyrene backbone and carbohydrate brushes. A large pseudo-helical polystyrene backbone is formed by a random sequence of TT, TG, and/or TTGG. The results indicate that the cross-section of a backbone chain with smaller oligosaccharide side-chains is obliged to expand more than that with longer side-chains. Even with rigid hydrophilic pendant oligosaccharide chains, the larger pseudo-helix of the main chain could orient the side-chains so as to envelop the hydrophobic backbone in aqueous solution. Thus the conformation of the main chain is determined not only by the chemical nature of an oligosaccharide chain but also by its length. PMID- 15780264 TI - A novel xyloglucan from seeds of Afzelia africana Se. Pers.--extraction, characterization, and conformational properties. AB - This paper is the first multi-scale characterization of the xyloglucan extracted from seeds of the African tree Afzelia africana Se. Pers. It describes the extraction and characterization of this polysaccharide in terms of both primary monosaccharide and oligosaccharide composition. It also includes a study of the seed morphology. Morphological characterization includes optical, transmission, and scanning electron microscopy. The polysaccharide exists in thickened cell walls of the cotyledonary cells, and the extracted xyloglucan is structurally quite similar to those from tamarind seed and detarium. Nevertheless there are some subtle differences in the fine structure, particularly in the oligomeric xyloglucan composition. The chain flexibility of the polysaccharide is also discussed in the light of our recent measurements reported elsewhere [Biomacromolecules2004, 5, 2384-2391]. PMID- 15780265 TI - Structural elucidation of type III group B Streptococcus capsular polysaccharide using molecular dynamics simulations: the role of sialic acid. AB - The conformational properties of the capsular polysaccharide (CPS) from group B Streptococcus serotype III (GBS III) are derived from 50 ns explicitly solvated molecular dynamics simulations of a 25-residue fragment of the CPS. The results from the simulations are shown to be consistent with experimental NMR homo- and heteronuclear J-coupling and NOE data for both the sialylated native CPS and for the chemically desialylated polysaccharide. A helical structure is predicted with a diameter of 29.3 A and a pitch 89.5 A, in which the sialylated side chains are arrayed on the exterior surface of the helix. The results provide an explanation for the observation that CPS antigenicity varies with carbohydrate chain length up to approximately 4 pentasaccharide repeat units. The conformation of the immunodominant region is established and shown to be independent of the presence of sialic acid. The data provide an explanation for the observation that the specificity of the determinant, associated with the major population of antibodies raised upon immunization of rabbits with GBS III, is dependent on the presence of sialic acid. In the sialylated native CPS, the antibody response is largely directed against the immunodominant core of the helix. From simulations of the desialylated CPS, a model emerges which suggests that the minor population of antibodies, whose determinant is not sialic acid dependent, recognizes the same immunodominant region, but that in the disordered CPS this region is not presented in a regular repeating motif. PMID- 15780266 TI - Conformation of the exopolysaccharide of Burkholderia cepacia predicted with molecular mechanics (MM3) using genetic algorithm search. AB - We present a computational conformational analysis of the exopolysaccharide of Burkholderia cepacia, which is believed to play a role in colonization and persistence of B. cepacia in the lungs of cystic fibrosis patients. The repeating unit of the exopolysaccharide is a heptasaccharide with three branches, which cause significant steric restraints. Conformational searches using glygal, an in house developed software using genetic algorithm search methods, were performed on fragments as well as on the complete repeating unit with wrap-over residues. The force field used for the calculations was MM3(96). The search showed four favored conformations for an isolated repeating unit. However, for a sequence of several repeating units, the calculations indicate a single, well-defined linear conformation. PMID- 15780267 TI - Conformational features of cepacian: the exopolysaccharide produced by clinical strains of Burkholderia cepacia. AB - Conformational energy calculations and molecular dynamics investigations, both in water and in dimethyl sulfoxide, were carried out on the exopolysaccharide cepacian produced by the majority of the clinical strains of Burkholderia cepacia, an opportunistic pathogen causing serious lung infection in patients affected by cystic fibrosis, The investigation was aimed at defining the structural and conformational features, which might be relevant for clarification of the structure-function relationships of the polymer. The molecular dynamics calculations were carried out by Ramachandran-type energy plots of the disaccharides that constitute the polymer repeating unit. The dynamics of an oligomer composed of three repeating units were investigated in water and in Me2SO, a non-aggregating solvent. Analysis of the time persistence of hydrogen bonds showed the presence of a large number of favourable interactions in water, which were less evident in Me2SO. The calculations on the cepacian chain indicated that polymer conformational features in water were affected by the lateral chains, but were also largely dictated by the presence of solvent. Moreover, the large number of intra-chain hydrogen bonds in water disappeared in Me2SO solution, increasing the average dimension of the polymer chains. PMID- 15780268 TI - A dynamic perspective on the molecular recognition of chitooligosaccharide ligands by hevein domains. AB - The complexes between hevein and different chitin oligomers, from the di- to the penta-saccharide, are studied through all atom molecular-dynamics simulations. The results for the smaller oligosaccharide complexes show that the carbohydrate is able to move on the surface of the relatively flat binding-pocket of hevein, therefore occupying different binding subpockets. The pentasaccharide spans all possible intermolecular interactions with the receptor in a simultaneous manner. Statistical analysis methods were also applied in order to define the principal overall motions in the complexes. The oligosaccharide binding can be considered to be defined by a subtle balance between enthalpic and entropic effects, providing the possibility of the existence of multiple binding conformations. This structural and dynamical view parallels the results based on NOE NMR data for the three disaccharide, trisaccharide, and pentasaccharide complexes. PMID- 15780269 TI - Potential transition-state analogs for glycosyltransferases. Design and DFT calculations of conformational behavior. AB - The structure of a previously calculated transition state (TS) was used to design the [tetrahydro-2-(methylthio)furan-2-yl]methyl phosphate dianion (1) as a new scaffold for transition-state analogs of reactions catalyzed by the inverting glycosyltransferases. This scaffold contains relevant features of the donor and acceptor and represents a new type of potential inhibitors for these enzymes. Available conformational space of 1 was explored using DFT quantum chemical methods by means of two-dimensional potential-energy maps calculated as a function of Phi, Psi, and omega dihedral angles at the B3LYP/6-31+G* level. The calculated potential energy surfaces revealed the existence of several low-energy domains. Structures from these regions were refined at the 6-311++G** level and led to 14 conformers. The stability of conformers is influenced by their environment, and in aqueous solution two conformers dominate the equilibrium. A superposition of calculated conformers with the predicted TS structure revealed that the preferred conformers in solution nicely mimic structural features of the TS. These results imply that 1 has structural properties required to mimic the TS and therefore can be used as a scaffold for further development of TS-analog inhibitors for retaining glycosyltransferases. PMID- 15780270 TI - The use of a genetic algorithm search for molecular mechanics (MM3)-based conformational analysis of oligosaccharides. AB - We have implemented a system called glygal that can perform conformational searches on oligosaccharides using several different genetic algorithm (GA) search methods. The searches are performed in the torsion angle conformational space, considering both the primary glycosidic linkages as well as the pendant groups (C-5-C-6 and hydroxyl groups) where energy calculations are performed using the MM3(96) force field. The system includes a graphical user interface for setting calculation parameters and incorporates a 3D molecular viewer. The system was tested using dozens of structures and we present two case studies for two previously investigated O-specific oligosaccharides of the Shigella dysenteriae type 2 and 4. The results obtained using glygal show a significant reduction in the number of structures that need to be sampled in order to find the best conformation, as compared to filtered systematic search. PMID- 15780271 TI - Surface chemical studies on the competitive adsorption of poly(ethylene glycol) and ammonium poly(methacrylate) onto alumina. AB - The adsorption of poly(ethylene glycol) (PEG) and ammonium poly(methacrylate) (APMA) onto alumina has been examined both individually and in combination. The adsorption density of APMA was found to be higher than that of PEG onto alumina. The adsorption isotherms of PEG and APMA for alumina exhibited a Langmuirian behavior. The adsorption density of PEG was significantly reduced in the presence of APMA, but the reverse was not true. About 60% desorption of PEG from alumina was achieved, while in the case of APMA the amount desorbed was only 10% in the pH range of 3-6. The zeta potential values of alumina were decreased and the isoelectric point (i.e.p.) values were shifted toward acidic pH values, proportional to the concentration of APMA added. However, such changes in the electrokinetic behavior were not observed by the addition of PEG. The dispersion behavior of alumina in the combined presence of PEG and APMA essentially followed the trends obtained for the alumina-APMA system, corroborating the electrokinetic measurements. Coprecipitation tests confirmed complexation between aluminum species and APMA in the bulk solution, but not with PEG. The interaction between alumina and PEG is primarily governed by hydrogen-bonding forces, while both hydrogen bonding and chemical interaction are involved in the case of the alumina APMA system. FTIR spectroscopic studies provided evidence in support of the interaction mechanisms proposed. PMID- 15780272 TI - The fabrication and characterization of a formaldehyde odor sensor using molecularly imprinted polymers. AB - The fabrication and characterization of odor sensors based on molecularly imprinted polymers is reported as the first case of imprinting formaldehyde. A quartz crystal microbalance is employed as a sensitive apparatus of a sensor for the determination of odor formaldehyde. An equation is deduced to characterize the interaction between molecularly imprinted films and the template. A linear relationship between the frequency shifts and the concentration of analyte in the range of 1.25 to 14.25 microM is found. The detection limit is about 20.5 microM. The sensor can selectively distinguish gaseous formaldehyde. It is envisaged that this novel and handy method could be employed to determine formaldehyde gas in the atmosphere. PMID- 15780273 TI - Surface complexation modeling of the sorption of 2-, 3-, and 4-aminopyridine by montmorillonite. AB - The sorption of 2-, 3-, and 4-aminopyridine on K-saturated Wyoming (SWy-K) and Texas (STx-K) and Ca-enriched Texas (STx-Ca) montmorillonite was measured at 25 degrees C with 10 mM KNO(3) or 3.3 mM Ca(NO(3))(2) as the background electrolyte. The aminopyridines adsorbed to montmorillonite at low pH, but not at high pH. Extended constant capacitance surface complexation models (ECCMs) and attenuated total reflectance-FTIR data indicate that aminopyridines sorb to the silica-like faces by cation exchange, forming outer-sphere complexes between aminopyridinium ions and permanent negatively charged surface sites (X(-)). X-ray diffraction data and sorption kinetics suggest that sorption occurs not only at external X(-) sites but also at those in the interlayer spaces. Differences in the sorption behaviors of 2-, 3-, and 4-aminopyridine result from differences in their pK(a)s. The extent of sorption of aminopyridines by the montmorillonite samples (SWy K>STx-K>STx-Ca) results from the higher cation-exchange capacity of SWy-K, and from the fact that Ca(2+) is much more effective than K(+) in competing with protonated aminopyridines for the X(-) sites. PMID- 15780274 TI - Effect of low-molecular-weight organic anions on exchangeable aluminum capacity of variable charge soils. AB - Low-molecular-weight (LMW) organic acids exist widely in soils and have been implicated in many soil processes, such as nutrient availability, translocation of metals, fate of heavy metals, and mineral weathering. In this paper, the effect of the LMW organic anions on the exchangeable aluminum of two variable charge soils was examined. The results showed that the organic anions induced an increase or a decrease in the exchangeable Al, and the extent and direction of the effect depended on the nature of organic anions, surface chemical properties of soils, and pH. For example, at pH 4.5, the quantity of exchangeable Al of Oxisol in the control system was 2.65 mmol kg(-1), whereas the values in the citrate, oxalate, malonate, malate, tartarate, salicylate, and lactate systems increased by 3.25, 1.93, 1.95, 1.82, 1.28, 0.88, and 0.45 times, respectively. In contrast, the quantity of the exchangeable Al of Ultisol at pH 4.5 in the oxalate and the citrate systems decreased by 8.8 and 19.6%, respectively. The increase in the exchangeable Al was caused mainly by the increase in negative surface charge of the soils due to the specific adsorption of organic anions. The ability of organic anions at low concentrations to increase exchangeable Al for Oxisol followed the order citrate > oxalate and malonate > malate > tartarate > salicylate > maleate > lactate. This order is consistent with that of the effect of the adsorption of anions on the increase in the negative surface charge and/or the decrease in the positive surface charge of the soil. On the other hand, the organic anions could depress the exchangeable Al through the formation of soluble Al-organic anion complexes under certain conditions. The anions with small stability constants of Al-organic anion complexes, such as lactate, caused an increase in exchangeable Al with the change in surface charge of the soils, while those with large stability constants, such as citrate and oxalate, caused an increase in the exchangeable Al at low concentration and a decrease at high concentration. PMID- 15780275 TI - Cosorption of Zn(II) and 2-, 3-, or 4-aminopyridine by montmorillonite. AB - Data from acid-base titrations at 25 degrees C of Zn(NO(3))(2) and 2-, 3-, or 4 aminopyridine in 10 mM KNO(3) as background electrolyte suggested that soluble complexes ZnL(2+) and Zn(OH)L(+) form, where L represents aminopyridine. Zinc hydroxyaminopyridine complexes have not been reported previously. The cosorption of Zn(II) with each of the aminopyridines to K-saturated Wyoming (SWy-K) and Texas (STx-K), and Ca-enriched Texas (STx-Ca) montmorillonites was measured at 25 degrees C, with 10 mM KNO(3) or 3.3 mM Ca(NO(3))(2) as background electrolyte. Comparison with previous data for sorption of Zn(II) and the aminopyridines separately and surface complexation modeling of the cosorption data showed that under acid conditions competition between Zn(2+) and aminopyridinium ions for the permanent negatively charged sites of montmorillonite results in suppression of the uptake of each sorbate by the other, but only when a large excess of the competing sorbate is present. Under alkaline conditions the sorption of Zn(II) was not affected by the presence of even a large excess of aminopyridine, but the sorption of 4-aminopyridine in particular was slightly enhanced when a large excess of Zn(II) was present. The enhancement was attributed to the formation of metal-bridged ternary surface complexes at the variable-charge sites on the edges of the montmorillonite crystals. PMID- 15780276 TI - Ammonium sorption from aqueous solutions by the natural zeolite Transcarpathian clinoptilolite studied under dynamic conditions. AB - The scope of this study is ammonium-ion uptake from synthetic aqueous solutions onto raw and pretreated forms of the natural zeolite Transcarpathian clinoptilolite under dynamic conditions. Hydrogen ions displaced exchangeable cations on the clinoptilolite in distilled water (sodium ions) and hydrochloric acid (sodium, potassium, and calcium ions) and destroyed the zeolite framework structure in the last case. Ammonium uptake onto the zeolite occurs by exchange with Na(+), Ca(2+), and K(+) ions. Although Na(+) ions were observed to be more easily exchanged for both hydrogen and ammonium ions, the role of Ca(2+) ions increased with zeolite saturation by NH(+)(4) ions. The maximum sorption capacity of the clinoptilolite toward NH(+)(4) ions, estimated under dynamic conditions, is significantly higher than that measured under static conditions; proximity of the values of a distribution coefficient and a retardation factor for different conditions (215-265 dm(3)/kg and 979-1107, respectively) allows us to use these parameters to model ammonium uptake onto the clinoptilolite. Slowing down or interruption in filtration resulted in the improvement of ammonium sorption properties of the zeolite. The ammonium removal improves with use of the finer fractions of the clinoptilolite up to 0.35 mm. A recycling study results confirmed the importance of external diffusion for ammonium sorption by the clinoptilolite. Preliminary treatment of the sorbent confirmed the predominant importance of the ion-exchange mechanism. The advantage of prior NaCl treatment of the clinoptilolite in improvement of ammonium removal over the other techniques was shown. PMID- 15780277 TI - Adsorption of lignite-derived humic acids on coal-based mesoporous activated carbons. AB - The adsorption by a coal-based mesoporous activated carbon of humic acids (HAs) isolated from two Polish lignites was studied. For comparison, a commercial Aldrich humic acid was also included into this study. The differences in chemical structure and functional groups of HAs were determined by elemental analysis and infrared spectroscopy DRIFT. Two activated carbons used differed in terms of mesopore volume, mesopore size distribution, and chemical properties of the surface. The kinetics of adsorption of HAs have been discussed using three kinetic models, i.e., the first-order Lagergren model, the pseudo-second-order model, and the intraparticle diffusion model. It was found that the adsorption of HAs from alkaline solution on mesoporous activated carbon proceeds according to the pseudo-second-order model. The correlation coefficients were close to 1. The intraparticle diffusion of HA molecules within the carbon particle was identified to be the rate-limiting step. Comparing the two activated carbons, the carbon with a higher volume of pores with widths of 10-50 nm showed a greater removal efficiency of HA. An increase in the Freundlich adsorption capacity with decreasing carbon content of HA was observed. Among the HAs studied, S-HA shows characteristics indicating the highest contribution of small-size fraction. The S HA was removed by both activated carbons to the highest extent. The effect of pH solution on the adsorption of HA was examined over the range pH 5.4-12.2. It was found that the extent of adsorption decreased with decreasing pH of the solution. PMID- 15780278 TI - A mathematical simulation of H+ ion chemisorption by anilinepropylsilica xerogels. AB - A study of the basic characteristics of the hybrid anilinesilica xerogel, which shows promising metal sorption properties, was carried out. A simplified approach to describe the protonization affinity of the immobilized aniline was developed, and it was compared with the known values for free aniline. The proton chemisorption process may be described properly by a model based on the Langmuir equation, and the immobilized groups have less affinity to the H(+) than to the free aniline. PMID- 15780279 TI - Adsorption equilibrium and transport kinetics for a range of probe gases in Takeda 3A carbon molecular sieve. AB - Measurements of adsorption equilibria and transport kinetics for argon, oxygen and nitrogen at 20, 50, and 80 degrees C on commercially derived Takeda carbon molecular sieve (CMS) employed for air separation have been undertaken in an effort to elucidate fundamental mechanisms of transport. Results indicate that micropore diffusion which is modeled by a Fickian diffusion process, governs the transport of oxygen molecules and the pore mouth barrier controls argon and nitrogen transport which is characterized by a linear driving force (LDF) model. For the three temperatures studied, the pressure dependence of the diffusivity and the LDF rate constant appear to be well characterized by a formulation based on the chemical potential as the driving force for transport. Isosteric heat of adsorption at zero loading and activation energy measurements are compared with predictions made from a previously proposed molecular model for characterizing CMS. PMID- 15780280 TI - The physical and surface chemical characteristics of activated carbons and the adsorption of methylene blue from wastewater. AB - Adsorption of a basic dye, methylene blue, from aqueous solutions onto as received activated carbons and acid-treated carbons was investigated. The physical and surface chemical properties of the activated carbons were characterized using BET-N(2) adsorption, X-ray photoelectron spectroscopy (XPS), and mass titration. It was found that acid treatment had little effect on carbon textural characteristics but significantly changed the surface chemical properties, resulting in an adverse effect on dye adsorption. The physical properties of activated carbon, such as surface area and pore volume, have little effect on dye adsorption, while the pore size distribution and the surface chemical characteristics play important roles in dye adsorption. The pH value of the solution also influences the adsorption capacity significantly. For methylene blue, a higher pH of solution favors the adsorption capacity. The kinetic adsorption of methylene blue on all carbons follows a pseudo-second-order equation. PMID- 15780281 TI - The effects of 1-hydroxyethane-(1,1-diphosphonic acid) on the adsorptive partitioning of metal ions onto gamma-AlOOH. AB - 1-Hydroxyethane-(1,1-diphosphonic acid) (HEDP) is extensively used in many technical applications. This work, as a first stage, examines the adsorption of aqueous HEDP, Cu(II), and Zn(II) onto boehmite (gamma-AlOOH) as single solutes. These processes can be described by the formation of inner-sphere complexes by the surface complexation model with constant capacitance. Uncomplexed HEDP is adsorbed as a mononuclear complex over the entre pH range. Four mononuclear surface complexes with different protonation levels are required to obtain a good fit of the experimental data for the pH range studied. Cu(II)- and Zn(II)-HEDP complexes at equimolar concentrations are studied at high and low surface density. The results indicate that the presence of HEDP significantly promotes metal adsorption at low pH in defects of surface sites. However, metal adsorption exhibits the same trend in the absence and presence of HEDP when the surface sites are in excess. The constant capacitance model successfully describes the experimental data through a ligand-like adsorption complex at low pH. At high pH, the model predicts separate adsorption of divalent metal and HEDP onto different surface sites as the preferred adsorption form. PMID- 15780282 TI - Salt softening of polyelectrolyte multilayer microcapsules. AB - By using a combination of atomic force and confocal microscopy, we explore the effect of 1:1 electrolyte (NaCl) on the stiffness of polyelectrolyte microcapsules. We study the "hollow" and "filled" (with polystyrene sulfonate) capsules. In both cases the shells are composed of layers of alternating polystyrene sulfonate (PSS) and polyallylamine hydrochloride (PAH). The stiffness of both "hollow" and "filled" capsules was found to be largest in water. It decreases with salt concentration up to approximately 3 mol/L and gets quasi constant in more concentrated solutions. The "filled" capsules are always stiffer than "hollow." The observed softening correlates with the salt-induced changes in morphology of the multilayer shells detected with the scanning electron microscopy. It is likely that at concentrations below approximately 3 mol/L the multilayer shell is in a "tethered" state, so that the increase in salt concentration leads to a decrease in number of ionic cross-links and, as a result, in the stiffness. In contrast, above the critical concentration of approximately 3 mol/L multilayer shells might be in a new, "melted," state. Here the multilayer structure is still retained, but sufficient amount of ionic cross links is broken, so that further increase in salt concentration does not change the capsule mechanics. These ideas are consistent with a moderate swelling of multilayers at concentrations below approximately 3 mol/L and significant decrease in their thickness in more concentrated solutions measured with surface plasmon spectroscopy. PMID- 15780283 TI - Size and shape of soil humic acids estimated by viscosity and molecular weight. AB - Ultrafiltration fractions of three soil humic acids were characterized by viscometry and high performance size-exclusion chromatography (HPSEC) in order to estimate shapes and hydrodynamic sizes. Intrinsic viscosities under given solute/solvent/temperature conditions were obtained by extrapolating the concentration dependence of reduced viscosities to zero concentration. Molecular mass (weight average molecular weight (M (w)) and number average molecular weight (M (n))) and hydrodynamic radius (R(H)) were determined by HPSEC using pullulan as calibrant. Values of M (w) and M (n) ranged from 15 to 118 x 10(3) and from 9 to 50 x 10(3) (g mol(-1)), respectively. Polydispersity, as indicated by M (w)/M (n), increased with increasing filter size from 1.5 to 2.4. The hydrodynamic radii (R(H)) ranged between 2.2 and 6.4 nm. For each humic acid, M (w) and [eta] were related. Mark-Houwink coefficients calculated on the basis of the M (w) [eta] relationships suggested restricted flexible chains for two of the humic acids and a branched structure for the third humic acid. Those structures probably behave as hydrated sphere colloids in a good solvent. Hydrodynamic radii of fractions calculated from [eta] using Einstein's equation, which is applicable to hydrated sphere colloids, ranged from 2.2 to 7.1 nm. These dimensions are fit to the size of nanospaces on and between clay minerals and micropores in soil particle aggregates. On the other hand, the good agreement of R(H) values obtained by applying Einstein's equation with those directly determined by HPSEC suggests that pullulan is a suitable calibrant for estimation of molecular mass and size of humic acids by HPSEC. PMID- 15780284 TI - Rheological properties of protein-surfactant based gels. AB - Water-based protein-surfactant gels, formed by mixing bovine serum albumin (BSA) and sodium dodecyl sulfate in water, were investigated by rheological methods. The measurements were performed for many different protein-to-surfactant ratios as a function of the applied frequency, stress, or strain, as well as by changing the temperature, in the range between 15 and 65 degrees C. The rheological behavior of the gels as a function of applied frequency is interpreted in terms of the overlapping of at least two viscoelastic relaxation processes. The rheological results indicate the presence of thermal transitions from essentially viscous to mainly elastic regimes, in analogy with the thermal gelation processes observed in polymer solutions. The thermal gelation threshold in the present system is modulated by the protein/surfactant ratio. Differential scanning calorimetry measurements were also performed to determine whether thermal gelation is somehow concomitant to protein denaturation. The results indicate that the thermal denaturation of BSA in protein-surfactant based gels occurs at slightly higher temperatures than in the bulk. Scanning electron microscopy indicates the occurrence in the gel structure of globules formed by the arrangement of fibrils. PMID- 15780285 TI - An evaluation of the colloidal stability of metal working fluid. AB - The effect of calcium on the stability of a commercial MWF is characterized through the experimental determination of the stability ratio, W. Three experimental methods of stability ratio evaluation are investigated. (1) The initial slope of the absorbance versus time curve is used to estimate the rate of coagulation. (2) Absorbance measurements are used to estimate N(0)/N with time. The stability ratio is determined from the slope of N(0)/N versus time. (3) Photon correlation spectroscopy (PCS) measurements of the volume distribution with time are used to estimate N(0)/N with time. Electrophoretic mobility was also measured and used to determine the fast coagulation concentration of the MWF. The accuracy of the experimentally determined stability ratios is evaluated using a population balance coagulation model. The model predicts the population distribution of a coagulating dispersion with time based on an initial particle size distribution and stability ratio. The model results were compared with the PCS-measured distributions to determine which stability ratio evaluation method best describes the stability of the MWF emulsion studied. Using the initial slope of the absorbance versus time curve to determine the fast coagulation concentration correlates well with electrophoretic mobility measurements. However, using absorbance measurements to determine the rate of coagulation underestimates the stability ratio of the MWF studied by orders of magnitude. N(0)/N values calculated from absorbance measurements provide a reasonable estimate of the stability ratio but inconsistencies in the method decrease its reliability. The stability ratio derived from PCS measurements appears to provide the most accurate, reliable description of MWF stability. PMID- 15780286 TI - Sonochemical synthesis of stable hydrosol of Fe3O4 nanoparticles. AB - The sonolysis of an aqueous solution of Fe(CO)(5) in the presence of sodium dodecyl sulfate leads to the formation of a stable hydrosol of amorphous Fe(3)O(4) nanoparticles. The amorphicity of iron oxide nanoparticles was determined by X-ray diffraction and differential scanning calorimetry. The nanoparticles were characterized by elemental analysis, EDX, transmission electron microscopy, dynamic light scattering, Raman spectroscopy, XPS, and spot test. PMID- 15780287 TI - Synthesis and characterization of CdS nanoparticles embedded in a polymethylmethacrylate matrix. AB - CdS nanopowder capped with sodium bis(2-ethylhexyl)sulfosuccinate was synthesized by using water-in-oil microemulsions. The CdS nanoparticles of about 5 nm obtained were embedded in polymethylmethacrylate matrix by a photocuring process. The transparent yellow solid compound was characterized by optical absorption and emission spectroscopy, high-resolution transmission electron microscopy, and energy-dispersive X-ray spectroscopy. The properties of this compound were compared with those of the nanopowder dispersed in heptane and in methylmethacrylate. The results obtained indicate that the nanoparticles are homogeneously dispersed in the matrix and do not change in size during the embedding process. Even if the surface slightly changes its luminescence properties, as a consequence of the different new chemical environment, the final product seems to be suitable for practical applications. PMID- 15780288 TI - Mobility of permeable fractal agglomerates in slip regime. AB - Hydrodynamic drag and mobility of fractal aggregates in the slip creeping flow regime are calculated. A theoretical continuum model of the gas slip flow past and within agglomerates is developed. It accounts for effects of flow rarefaction and porous fractal structure upon the molecular mean free path, apparent viscosity, and effective permeability of agglomerates. It is shown that flow rarefaction significantly diminishes the aggregates' drag to an extent that cannot be predicted by the Cunningham's drag correction factor. The developed model allows calculation the agglomerates' transport properties in a wide range of fractal dimensions. For low D(f) agglomerates the drag force agrees with the Friedlander's expression based on the Epstein's single sphere drag in the free molecular regime. PMID- 15780289 TI - Size-controlled preparation of Cu2O octahedron nanocrystals and studies on their optical absorption. AB - We report herein the size-controlled preparation of monodispersed cuprous oxide octahedron nanocrystals smaller than 100 nm. The method is based on the reduction of copper nitrate in Triton X-100 water-in-oil (w/o) microemulsions by gamma irradiation. The average edge length of the octahedron-shaped nanocrystals varies from 45 to 95 nm as a function of the dose rate. The quantum confinement effect was illustrated by the blueshift in the optical absorption. In addition, the growth process was also traced by absorption spectra. PMID- 15780290 TI - Effect of layer thickness on the luminescence properties of ZnS/CdS/ZnS quantum dot quantum well. AB - ZnS/CdS/ZnS quantum dot quantum well was prepared. The optical properties of ZnS/CdS/ZnS QDQW with different thickness of CdS well and ZnS shell were studied. Absorption spectra, emission spectra, and luminescence lifetimes were measured. The observed luminescence was assigned to the bulk donor-acceptor pair recombination of CdS and can be enhanced by increasing the thickness of the CdS well or coating an appropriate thickness of ZnS shell on the surface of the CdS well. The luminescence enhancement was caused by the relative reduce in the surface effect. The luminescence lifetimes were influenced strongly by the surface state. PMID- 15780291 TI - Synthesis of spherical silver nanoparticles by digestive ripening, stabilization with various agents, and their 3-D and 2-D superlattice formation. AB - Capped nanoparticles of silver were synthesized via the solvated metal atom dispersion (SMAD) technique followed by a digestive ripening procedure producing gram quantities of monodisperse spherical nanoparticles. This shows for the first time that a digestive ripening protocol is possible for an element other than gold. The particle size and optical spectra were found to be dependent on the capping agent used. Particles capped with dodecane thiol had a mean diameter of 6.6+/-1 nm, while trioctyl phosphine capped particles were 6.0+/-2 nm determined via TEM microscopy. These particles were found to organize into two- and three dimensional superlattices with a well defined geometry through self-assembly in a liquid solution, that was dictated by the ligand used resulting in a triangular or circular lattice. PMID- 15780292 TI - Stabilization of magnetorheological suspensions by polyacrylic acid polymers. AB - This work is devoted to the synthesis and stabilization of magnetorheological suspensions constituted by monodisperse micrometer-sized magnetite spheres in aqueous media. The electrical double-layer characteristics of the solid/liquid interface were studied in the absence and presence of adsorbed layers of high molecular weight polyacrylic acids (PAA; Carbopol). Since the Carbopol-covered particles can be thought of as "soft" colloids, Ohshima's theory was used to gain information of the surface potential and the charge density of the polymer layer. The effect of the pH of the solution on the double-layer characteristics is related to the different conformations of the adsorbed molecules provoked by the dissociation of the acrylic groups present in polymer molecules. The stability of the suspensions was experimentally studied for different pH and polymer concentrations, and in the absence or presence of a weak magnetic field applied. The stability of the suspensions was explained using the classical DLVO theory of colloidal stability extended to account for hydration, steric, and magnetic interactions between particles. Diagrams of potential energy vs interparticle distance show the predominant effect of steric, hydrophilic/hydrophobic, and magnetic interactions on the whole stability of the system. The best conditions to obtain stable suspensions were found when strong steric and hydrophilic repulsions hinder the coagulation between polymer-covered particles, simultaneously avoiding sedimentation by the thickening effect of the polymer solution. When a not too high molecular weight PAA was employed in a low concentration, the task of a long-time antisettling effect compatible with the desired magnetic response of the fluid was achieved. PMID- 15780293 TI - Effect of base addition rate on the preparation of partially neutralized ferric chloride solutions. AB - Cationic polymers prepared from partially neutralized ferric chloride solutions were investigated using photon correlation spectroscopy (PCS) techniques. To study the speciation and yield of the polymers, FeCl(3) solutions were hydrolyzed by adding NaHCO(3) solution for a range of hydrolysis ratios (B=[OH]/Fe(T)) of 0 to 2.0 under different base addition rates. It was observed that the prepared solution has a higher degree of polymerization at a higher base addition rate. The "B rate" (R(B), 1/time), i.e., the rate of change of B during preparation, was found to have a dramatic effect on the speciation of the partially neutralized ferric chloride solution. High-yield inorganic iron(III) polymer ( approximately 80%) can be produced in a specific range of B rates. These iron(III) polymers, having a size ranging from 3 to 7 nm, were stable without precipitation of ferric hydroxide microcolloids after aging for 2 weeks. PMID- 15780294 TI - Influence of nonwetting on the aggregation dynamics of micronic solid particles in a turbulent medium. AB - The aim of this work was to determine and to interpret the influence of nonwetting on the aggregation dynamics of micronic solid particles in a turbulent medium. Two silica granular samples were studied: one was naturally hydrophilic; the other was made hydrophobic. Aggregation in an aqueous ethanol solution was followed by in situ turbidimetry. The influence of stirring rate and deaeration was determined. Aggregates of hydrophilic particles were small and fragile, whereas aggregates of hydrophobic particles were large and solid. Moreover, they differred greatly in optical properties. Within the proposed approach, different features of the aggregate morphology were identified: fractal dimension, maximum size, and gas content of the hydrophobic clusters. These elements are taken into account in the models of aggregation dynamics proposed here. PMID- 15780295 TI - Interactions of hairy latex particles with cationic copolymers. AB - Interactions between polycations and core-corona particles are governed by ion exchange reactions, entropically favored by the release of counterions. This complexation process allows the chains to penetrate into the shell, leading to adsorbed amounts greater than 1 mg m(-2). The destabilization occurs quickly, the domain of flocculation becomes larger when the concentration of monovalent salts is increased, and aggregates are composed of small and very compact clusters in a more or less self-similar structure at large scale. The adsorption of copolymers of low cationicity is characterized by still larger adsorbed amounts and layers thicker than the radius of gyration of the macromolecules. Depending on the charge content, the enhancement of the ionic strength can either promote the destabilization of the suspension or conversely induce the desorption of the chain. In pure water the structure of the flocs is long-range ordered and it becomes more heterogeneous in ionic media. PMID- 15780296 TI - On the self-similar solution of fragmentation equation: Numerical evaluation with implications for the inverse problem. AB - It is well known that the fragmentation equation admits self-similar solutions for evolving particle-size distributions (PSD); i.e., if the shape of PSD is independent of time after an initial transient period. Although an analytical derivations of the self-similar PSD cases have been studied extensively, results for cases requiring numerical solutions are rare. The aim of the present work is to fill this gap for the case of homogeneous breakage functions. The known analytical and approximate solutions for the self-similar PSD are reviewed and a general algorithm for the numerical solution is proposed. Results for a broad range of breakage functions (kernel and rate) are presented. Further, the work is focused on the sensitivity of the relation between self-similar PSD and breakage kernel and its influence on the inverse breakage problem, i.e., that of estimating the breakage kernel from experimental self-similar PSDs. Useful suggestions are made for tackling the inverse problem. PMID- 15780297 TI - Effects of a central metal on the organization of 5,10,15,20-tetra-(p chlorophenyl)-rare earth porphyrin hydroxyl compound at the air/water interface and in Langmuir-Blodgett films. AB - Langmuir monolayers and Langmuir-Blodgett (LB) films of 5,10,15,20-tetra-(p chlorophenyl) terbium/gadolinium porphyrin hydroxyl compound (TbOH and GdOH) and their mixtures with stearic acid (SA) in a molar ratio of 1:1 were investigated by Brewster angle microscopy (BAM), ultraviolet-visible (UV-vis), and infrared (IR) spectroscopy and atomic force microscopy (AFM). pi-A isotherms showed that well-defined Langmuir monolayers were formed at an air/water interface for the porphyrins and their mixture with SA. The BAM observations suggest that the pi-pi interaction between the GdOH molecules is stronger than that between the TbOH molecules. This result can be further confirmed by the AFM measurements. After the introduction of SA, the pi-pi interaction between the TbOH molecules is broken and thus two phases formed in the mixed LB film. However, it cannot break the stronger pi-pi interaction between the GdOH molecules. Therefore, no phase separation is observed in the GdOH/SA LB film. IR reflection-absorption (RA) spectra showed that the COOH groups of SA are partly converted to COO(-) groups, suggesting that there is an interaction between MOH and SA in the films. This interaction leads the benzene rings of TbOH to rotate toward parallel to the substrate and those of GdOH to rotate toward perpendicular to the substrate. All these results have demonstrated that the central metal ions have great effects on the organization and formation of the films. PMID- 15780298 TI - Carbon nanotubes as a secondary support of a catalyst layer in a gas diffusion electrode for metal air batteries. AB - In this paper, we report the use of binary carbon supports (carbon nanotubes (CNTs) and active carbon) as a catalyst layer for fabricating gas diffusion electrodes. The electrocatalytic properties for the oxygen reduction reaction (ORR) were evaluated by polarization curves and electrochemical impedance spectroscopy (EIS) in an alkaline electrolyte. The binary-support electrode exhibits better performance than the single-support electrode, and the best performance is obtained when the mass ratio of carbon nanotubes and active carbon is 50:50. The results from the electrode kinetic parameters indicate that the introduction of carbon nanotubes as a secondary support provides high accessible surface area, good electronic conductivity, and fast ORR kinetics. Furthermore, the effect of CNT support on the electrocatalytic properties of Pt nanoparticles for binary-support electrodes was also investigated by different loading reduction methods. The electrocatalytic activity of the binary-support electrodes is improved dramatically by Pt loading on CNT carbon support, even at very low Pt loading. Additionally, the EIS analysis results indicate that the process of ORR may be controlled by diffusion of oxygen in the electrode thin film for binary support electrodes with or without Pt catalyst. PMID- 15780299 TI - Adsorption and redox reactions of heavy metals on Fe-Mn nodules from Chinese soils. AB - Adsorption of heavy metals and redox reactions of Cr(W) ions on Fe-Mn nodules from five soils of China were investigated by chemical analysis, equilibrium adsorption/redox, and X-ray photoelectron spectroscopy (XPS). Results show that Mn is mainly present as Mn(3+) and Mn(4+) forms in Fe-Mn nodules. The maximum adsorption amounts for different heavy metal ions follow the order Pb(2+) approximately Cu(2+)>Zn(2+)>Co(2+)>Ni(2+)>Cd(2+). The adsorption capacity for heavy metals by Fe-Mn nodules from calciaquert in Shandong province (N5-1) is the highest, while that from hapludalf in Shandong province (N6-1) is the lowest. About 44-100% of the heavy metals adsorbed on Fe-Mn nodules were dissolved in 0.1 mol/L hydroxylamine hydrochloride (HAHC). The maximum amounts of Cr(VI) production by Fe-Mn nodules follow the order of N1-1 (69 mmol/kg)>N4-1 (57 mmol/kg)>N2-1 (52 mmol/kg)>N5-1 (44 mmol/kg). Based on the content of MnO(2) in Fe-Mn nodules dissolved in HAHC, the amount of Cr(VI) production by Mn oxides in N1-1, N2-1, N4-1, and N5-1 is 326, 624, 726, and 482 mmol/kg (MnO(2)), respectively. We propose that the amounts of Cr(VI) production through oxidation Cr(III) by Mn oxides are related to the types of Mn oxides in Fe-Mn nodules. PMID- 15780300 TI - Catalytic dehydration of ethanol using transition metal oxide catalysts. AB - The aim of this work is to study catalytic ethanol dehydration using different prepared catalysts, which include Fe(2)O(3), Mn(2)O(3), and calcined physical mixtures of both ferric and manganese oxides with alumina and/or silica gel. The physicochemical properties of these catalysts were investigated via X-ray powder diffraction (XRD), acidity measurement, and nitrogen adsorption-desorption at 196 degrees C. The catalytic activities of such catalysts were tested through conversion of ethanol at 200-500 degrees C using a catalytic flow system operated under atmospheric pressure. The results obtained indicated that the dehydration reaction on the catalyst relies on surface acidity, whereas the ethylene production selectivity depends on the catalyst chemical constituents. PMID- 15780301 TI - Evaluation of thermoporometry for characterization of mesoporous materials. AB - The accuracy of thermoporometry (TPM) in terms of the characterization of SBA-15 is examined based on a model that classifies the water in the mesopores into two different types: freezable pore water, which can form cylindrical ice crystals, and nonfreezable pore water, which cannot undergo a phase transition during a differential scanning calorimetry (DSC) measurement. Applying the empirical relationship between the sizes of the ice crystals formed in the mesopores and the solidification temperature of the freezable pore water to a thermogram (a recording of the heat flux during the solidification of the freezable pore water) yielded a size distribution of the ice crystals. The size of the ice crystals increased slightly with repetitive freezing, indicating that the mesopores were enlarged by formation of the ice crystals. Adding the thickness, t(nf), of the nonfreezable pore water layer to the ice crystal-size distribution calculated from the thermogram allowed for the determination of the porous properties of SBA 15. The porous properties attained from TPM experiments were compared with the results attained through the combination of Ar gas adsorption experiments and nonlocal density functional theory (NLDFT) analysis. The porous properties determined by TPM were confirmed to be quite sensitive to the t(nf) value. PMID- 15780302 TI - Theory of mercury intrusion in a distribution of unconnected wedge-shaped slits. AB - Effective mercury intrusion in a wedge-shaped slit is gradual, the intruded depth increasing with applied pressure. The Washburn equation must be modified accordingly. It relates the distance, e, separating the three-phase contact lines on the wedge faces to the hydrostatic pressure, P, wedge half-opening angle alpha, mercury surface tension gamma, and contact angle theta: e=( 2gamma/P)cos(theta-alpha) if theta-alpha>pi2. The equations relating the volume of mercury in a single slit to hydrostatic pressure are established. The total volume of mercury V(Hg)(tot)(E(0),e) intruded in a set of unconnected isomorphous slits (same alpha value) with opening width, E, distributed over interval [E(0),0], and volume-based distribution of opening width, f(V)(E), is written as where G(X)=(sin(-1)X-X1-X(2))/X(2) and X(E,e)=-cos(theta-alpha)Ee. The exact relation between total internal surface area and integral pressure work is. PMID- 15780303 TI - Quantifying the effect of ionic strength on colloidal fouling potential in membrane filtration. AB - Ultrafiltration experiments were conducted to study the fouling potential of colloidal suspensions under different ionic strengths and colloid concentrations. A linear relationship was found relating the colloidal fouling potential to the logarithm of the Debye-Huckel parameter, a characteristic for electrical double layers of colloids. This finding provided a useful quantitative linkage between the colloidal fouling potential and the water chemistry. Considering the linear dependence of colloidal fouling potential on the colloid concentration, a bilinear model was proposed to explain the coupling effects of colloid concentration and ionic strength of the suspension on the fouling potential. The model predictions of fouling potential were found to fit accurately with experimentally determined fouling potential values. Further analysis of the model showed that ionic strength can significantly affect colloidal fouling, for example, a 10-fold increase in ionic strength from 0.001 to 0.01 M for a given feed concentration has the same membrane fouling effect as doubling the feed concentration. The model allows for a quick and reliable assessment of fouling potential without even performing any experiments. This could then be used to design the membrane process or pretreatment stages required to mitigate membrane fouling. PMID- 15780304 TI - Characterization of latex particle arrays by gas adsorption. AB - Assemblies of colloidal particles are frequently used in novel applications, and this requires nondestructive methods allowing overall characterization of the sample and collection of information about the quality of the arrays. From suspensions of polystyrene, poly[styrene-co-(2-hydroxyethylmethacrylate)], poly[styrene-co-acrylic acid], and poly[styrene-co-methacrylic acid], assemblies of spherical particles were obtained by elimination of the solvent in different ways-evaporation, gravity deposition, and filtration. These latex particle packings were characterized by scanning and transmission electron microscopy and by gas adsorption to determine the efficiency of packing. The surface area, total pore volume, and pore size distributions obtained from the adsorption and desorption data were related to characteristic parameters calculated for cubic close-packed spherical particles. PMID- 15780305 TI - Synthesis of functional microcapsules containing suspensions responsive to electric fields. AB - A sort of functional microcapsules, which contain a suspension responsive to electric fields, is prepared by in situ polymerization of urea and formaldehyde. The suspension is made up of pigment phthalocyanine green (PPG) and tetrachloroethylene. In order to solve the particles' separation from the suspension during the microencapsulation and to obtain microcapsules applying to electronic ink display, the dispersibility of the particles, the contact angles between the particles and the tetrachloroethylene, and the influences of different emulsifiers on the microencapsulation are investigated. It is found that the dispersion extent and lipophilicity of the PPG particles are improved due to their surface modification with octadecylamine. The contact angles between the modified PPG particles and the tetrachloroethylene increase, and the PPG particles modified with 2 wt% octadecylamine have the best affinity for tetrachloroethylene. The interfacial tension between C(2)Cl(4) and H(2)O with urea-formaldehyde prepolymer descends from 43 to 35 mN/m, which indicates that the polymer has certain surface activity. However, water-soluble emulsifiers have an important influence during the microencapsulation because they can absorb on the surfaces of internal phase and prevent the resin of urea-formaldehyde from depositing there. From the SEM images of shell surface and cross section, the microcapsules have relatively smooth surfaces and the average thickness is about 4.5 mum. When the microcapsules are prepared with agitation rates of 1000 and 600 rpm, the mean diameters of the obtained microcapsules are 11 and 155 mum, respectively. The particles in the capsules move toward positive electrode with a responsive time of several hundred milliseconds while providing an electric field. PMID- 15780306 TI - Wetting films of polar and nonpolar liquids. AB - Disjoining pressure isotherms for water and n-octane films on glass obtained using a recently developed apparatus, based on the interferometric technique, are compared with other values reported in the literature. Our method, including a simple cleaning procedure, yields experimental results in excellent agreement both with other experimental results and with theoretical predictions. An error analysis was made which quantifies the uncertainty of our results and simultaneously allows the minimization of the errors. PMID- 15780307 TI - Self-aggregation of binary mixtures of alkyltriphenylphosphonium bromides: a critical assessment in favor of more than one kind of micelle formation. AB - The micellization behavior of binary combinations of alkyltriphenylphosphonium bromides (ATPBs) with alkyl chain carbons 10, 12, 14, and 16 has been studied by conductometry and calorimetry. The combinations C(10)-C(12), C(10)-C(14), C(10) C(16), C(12)-C(14), C(12)-C(16), and C(14)-C(16) were found to form two cmc's by both the methods, with good agreement, except C(14)-C(16)TPB, which has evidenced only a single cmc by calorimetry for all combinations. The combinations C(10) C(12) (for both cmc(1) and cmc(2)) and C(10)-C(14)TPB (for cmc(2)) formed ideal mixtures, whereas the rest were nonideal. In the nonideal binary mixtures, the ATPB components showed antagonistic interaction with each other. The cmc, interaction parameter (beta), mixed micellar composition, extent of counterion binding, and thermodynamic parameters for the micellization process have been reported and discussed. The enthalpy of mixed micelle formation has been found to have a fair correlation with a Clint-type relation applicable to ideal binary mixtures of surfactants. PMID- 15780308 TI - Interaction between cationic surfactants and montmorillonites under nonequilibrium condition. AB - Surfactant adsorption by two different montmorillonites was characterized by examining the time dependence of surfactant behavior on clay surfaces. Surfactants with different micelle concentrations were conducted in our experiment to observe a nonequilibrium activity of cationic surfactant on the clay over reaction periods ranging from 0.1 min to 11 days. Compared with Ca montmorillonite (SAz), a more active intrusion of surfactant molecules into the interlayers was found in Na-montmorillonite (SWy). During a short "initiation" stage, the basal spacing of SWy montmorillonite increased rapidly with logarithmic time. For SAz montmorillonite, however, the abrupt basal spacing increase occurred at a later stage of the reaction. From the results, it is assumed that the difference in the adsorption behavior exhibited by the two montmorillonite types partly arises from their intrinsic nature; that is, inorganic cations originally existed on the clay surfaces. Additionally, the micelle concentration of the surfactants affects the development of organomontmorillonite, especially in the intercalant formation and stabilization under nonequilibrium. PMID- 15780309 TI - Effect of AOT on enzymatic activity of the organic solvent resistant tyrosinase from Streptomyces sp. REN-21 in aqueous solutions and water-in-oil microemulsions. AB - The effect of AOT (sodium-bis(2-ethylhexyl sulfosuccinate)) on enzymatic activity of the organic solvent resistant tyrosinase (OSRT) in aqueous phosphate buffer solutions and in water-in-oil microemulsions of the water/AOT/isooctane system has been investigated. In contrast to mushroom tyrosinase, AOT does not activate OSRT in aqueous solutions, altering its activity very little at concentrations lower than 2 mM. Increasing contents of AOT in isooctane reduce the observed initial reaction rates of oxidation of t-butylcatechol (tBC) and 4-methylcatechol (4-MC). Similarly to mushroom tyrosinase, the effect has been described using an equation based on preferential binding of the substrates by surfactant interface layers. The apparent Michaelis-Menten substrate binding constants increase linearly with AOT concentration (with slopes of 0.12+/-0.02 and 0.051+/-0.006 for tBC and 4-MC, respectively), and the effective enzyme turnover number in the microemulsions remains practically constant. PMID- 15780310 TI - Fluorescence studies of interactions of ionic surfactants with poly(amidoamine) dendrimers. AB - The pyrene fluorescence measurements have been carried out for the micelle formation of sodium dodecyl sulfate (SDS), dodecyltrimethylammonium bromide (DTAB), and dimethylene bis(dodecyldimethylammonium bromide) (12-2-12) in the presence of fixed different amounts of various generations of poly(amidoamine) (PAMAM). The critical micelle concentration (cmc) of SDS decreases with an increase in the fixed amount of PAMAM, suggesting the facilitation of micellization due to the participation of SDS-PAMAM complex in the micelle formation. This behavior has not been observed for DTAB/12-2-12 in the presence of various generations of PAMAM. The results indicate that SDS always has stronger interactions with all the generations of PAMAM in comparison to those of DTAB and 12-2-12. PMID- 15780311 TI - Physical chemistry of nanostructured molecular sieves by the study of phase diagrams: the case of the cetyltrimethylammonium bromide-tetramethylammonium silicate-water system. AB - A phase diagram for the system cetyltrimethylammonium bromide (CTAB)/tetramethylammonium silicate (TMASi)/water has been constructed in order to better understand the interactions between these precursors of the MCM-41 mesoporous molecular sieves. Three different CTAB concentration regions were analyzed: the dilute and semidilute regions, where simple surfactant species, such as monomers and spherical and nonspherical CTAB micelles, are found, and the concentrated region, involving liquid-crystalline phases. In the dilute and semidilute regions, the formation of a white nanostructured solid, having a hexagonal array similar to that found in MCM-41 materials, was observed. Precipitation of this solid requires some degree of surfactant monomer aggregation, which is favored by the presence of silicate anions. If micelles have already been formed, the material can be obtained at any CTAB concentration above a threshold concentration of silicate anions. These facts suggest that silicate anions have an important role in changing the aggregation and/or the shape of the surfactant aggregates. In the concentrated region, precipitation of the solid was not observed, but the presence of the silicate anions alter the characteristics of the liquid-crystalline phase formed by the surfactant. The system shows very complex and rich behavior and its investigation may be very useful in understanding the processes of nanostructured solid formation. PMID- 15780312 TI - Molecular modeling study on the relative stabilities of the flotation products for arsenic-containing minerals: dixanthogens and arsenic(III) xanthates. AB - The interactions of As(III) ion with C(2)H(5)OCS(-)(2) and C(2)H(5)SCS(-)(2), known as the most popular collector ions, were studied by density functional theory (DFT) at the B3LYP/6-31G** level in connection with the arsenic-containing minerals realgar, orpiment, and arsenopyrite. The dixanthogen formations of these ions were also investigated at the same level of theory. The central purpose of this paper is to compare the stabilities of the major flotation products, namely As(III) xanthates and dixanthogens. The results show that the magnitudes of the interaction energies for the formations of As(III) xanthates and dixanthogens increase when the oxygen atom is replaced by the sulfur atom in C(2)H(5)OCS( )(2). Therefore, the C(2)H(5)OCS(-)(2) ion is preferred in these formations. The results obtained are in agreement with the experimental data reported. PMID- 15780314 TI - Structures of micelles formed by synthetic alkyl glycosides with unsaturated alkyl chains. AB - Three new alkyl glycosides with similar molecular structures (oleyl and oleoyl alkyl chains and various head groups: disaccharide, trisaccharide and disaccharide with an additional amidoethoxy spacer) were synthesized and their supramolecular structure in aqueous solution was investigated. Small angle neutron scattering, surface tension measurement and the contact preparation method were applied to get molecular structure-property relationships. Although the chemical structures differ only in small details, their CMC values, lyotropic phase behaviour, surface area per surfactant molecule in the micelle and at the liquid-air interface, and the size and shape of the micelles are very different. We have found three different types of aggregates: spherical, cylindrical and polymer-like micelles in dilute solutions. PMID- 15780313 TI - Transition from micelle to vesicle in aqueous mixtures of anionic/zwitterionic surfactants studied by fluorescence, conductivity, and turbidity methods. AB - Vesicles form spontaneously in a aqueous mixture of sodium bis(2-ethylhexyl) sulfosuccinate (Aerosol OT) and lauryl sulfonate betaine (LSB). Different from catanionic vesicles, the formation or disaggregation of such zwitterionic/anionic vesicles may be easily controlled by adjusting the relative amount of LSB and salinity. The participation of LSB reduces the polydispersity of the vesicles and even results in the formation of monodispersed vesicles at a certain salinity. But as LSB exceeds a certain proportion, vesicles cannot form at any concentration and salinity, making convenient the study of the structural transitions. We applied pyrene as a fluorescence probe and monitored the transition among the monomer, micelle, and vesicle through the variation of I(1)/I(3), accompanied by conductivity and turbidity measurements. In LSB solution and LSB-rich mixture, an abrupt change of the ratio of I(1)/I(3) was found in the transition from monomer to micelle with increasing concentration, as well as in the transition from micelle to vesicle with increasing salinity, which shows that a difference of the polarity of the microenvironment between the micelle and the vesicle bilayer resulted from the composition change. But in AOT solution and AOT-rich mixture, only a gradual change in the transition is observed due to the existence of intermediate structures, which have different microenvironments from micelles and vesicles. So the formation of vesicle experiences a process of monomer to premicelle to micelle to bilayer segment with increasing concentration by combining the conductivity method. The ratio of I(1)/I(3) is independent of the vesicle size once formed. PMID- 15780315 TI - Microstructural evolution of viscoelastic emulsions stabilised by sodium caseinate and xanthan gum. AB - The time-dependent evolution of the phase-separated microstructure of a caseinate stabilised emulsion containing xanthan gum added before emulsification has been investigated by confocal laser scanning microscopy, image analysis and rheology. Moderately low levels of xanthan addition lead to depletion flocculation and gravity-induced phase separation. Increasing the polysaccharide concentration causes immobilisation of the microstructure due to an increase in the local viscoelasticity: that is, the emulsion structure cannot easily rearrange to expel xanthan-enriched aqueous serum phase because a weak gel-like network is generated. The effect of xanthan on the evolving microstructure of phase separated regions, which reflects indirectly the local emulsion micro-rheology, has been estimated from image analysis of time sequences of confocal micrographs. A comparison has been made between object shape analysis using four different shape descriptors. The roundness parameter has been found to be a convenient descriptor for reliably quantifying the structural change in terms of the relaxation rate of xanthan-rich aqueous drops. The Taylor parameter has been used to link the kinetics of drop relaxation to the time-dependent small-deformation rheological behaviour. The analysis of the combined experimental data reveals the difficulty of relating the evolving microstructure to bulk rheological measurements. PMID- 15780316 TI - Drag force on a rigid spheroidal particle in a cylinder filled with Carreau fluid. AB - The boundary effect on the drag acting on a rigid particle is investigated by considering a spheroid on the axis of a cylinder filled with a Carreau fluid. The result of numerical simulation reveals that the ratio (drag coefficient in Carreau fluid/drag coefficient in Newtonian fluid) has a maximum as the ratio (semiaxis in radial direction/radius of cylinder) varies. The presence of a wall has the effect of enhancing the convective motion in the rear part of a particle, and therefore, the formation of wakes. The influence of the shape of a particle on the drag force acting on it can be decreased either by increasing the shear thinning effect of the fluid or by increasing the Reynolds number. The Reynolds number at which flow separation occurs is found to increase roughly linearly with the increase in the power-law exponent of the Carreau fluid. PMID- 15780317 TI - Shear-modulated electroosmotic flow on a patterned charged surface. AB - The effect of imposing shear flow on a charge-modulated electroosmotic flow is theoretically investigated. The flow structures exhibit either saddle points or closed streamlines, depending on the relative strength of an imposed shear to the applied electric field. The formation of closed streamlines could be advantageous for trapping nondiffusive particles at desired locations. Different time periodic alternating flows and their corresponding particle trajectories are also examined to assess strategies for creating efficient mixing. PMID- 15780318 TI - Timescales for relaxation to Boltzmann equilibrium in nanopores. AB - In most models for electrokinetic phenomena at charged interfaces, Boltzmann equilibrium is assumed to be established. Here we show that a long nanopore with significant double layer overlap establishes equilibrium quite slowly and that centimeter-long nanopores can take O(10(5)) s to establish Boltzmann equilibrium. The timescale is determined not by diffusion across the double layer, but by diffusion or convective transport along the length of the pore to reservoirs at its ends. An "intermediate equilibrium" state described by Qu and Li (J. Colloid Interface Sci. 224 (2000) 397) may exist for times between the (fast) EDL establishment timescale and (slow) axial transport timescale. PMID- 15780319 TI - Microcalorimetric study on the interaction of dissymmetric gemini surfactants with DNA. AB - The interaction of a series of dissymmetric gemini surfactants, [C(m)H(2m+1)(CH(3))(2)N(CH(2))(6)N(CH(3))(2)C(n)H(2n+1)]Br(2) (designated as C(m)C(6)C(n)Br(2), with constant m+n=24, and m=12, 14, 16, and 18) with DNA in 10 mM NaCl solution has been investigated by isothermal titration microcalorimetry (ITC). The curves for titration of the surfactants into DNA solution show noticeable differences from those into 10 mM NaCl solution without DNA. It is attributed to the interaction between DNA and surfactants. The critical aggregation concentration (CAC), the saturation concentration (C(2)), and the thermodynamic parameters for the aggregation and interaction processes were obtained from the calorimetric titration curves. The results show that the dissymmetry degree (m/n) has a marked effect on the interaction of the C(m)C(6)C(n)Br(2) surfactants with DNA. The CAC and C(2) tend to become smaller with increased m/n. The enthalpy change (DeltaH(agg)) and the Gibbs free energy change (DeltaG(agg)) for aggregation become more negative down the series, indicating that the hydrophobic interaction between the hydrophobic chains of the surfactant molecules increases and the aggregation process is more spontaneous with increased m/n. The entropy changes of aggregation (DeltaS(agg)) are all positive and TDeltaS(agg) is much larger than |DeltaH(agg)|, revealing that the aggregation process is mainly entropy-driven. However, the calculated Gibbs free energy (DeltaG(DS)) for the interaction between the gemini surfactants and DNA becomes less negative with increased m/n, which reveals that the interaction between the gemini surfactants and DNA tends to be weaker with increased m/n. This is induced by the disruption of the chain-chain hydrophobic interaction between the surfactant molecules at higher m/n, where the entropy change DeltaS(DS) for the interaction process tends to be an unfavorable factor. In addition, the DNA concentration also has a remarkable influence on the interaction. PMID- 15780320 TI - The Asian Tsunami remembered. PMID- 15780321 TI - Community-based public health interventions in North Korea: one non-governmental organization's experience with tuberculosis and hepatitis B. AB - BACKGROUND: The Democratic People's Republic of Korea, also known as North Korea, is the most isolated country in the world, and has been a source of interest, politically and socially, since the 1953 ceasefire of the Korean War. While in the past year, interest has focused on the nuclear dispute with the USA, over the past decade, most interest has revolved around the economic plight of the country, the lack of funds and resources for health care, and the subsequent public health declines. North Korea's present economic and public health problems began in the early 1990s due to the gradual loss of economic support from its communist allies (i.e. after the fall of the Soviet Union and the capitalization of China), combined with an inordinate number of natural disasters (floods, famine and drought) all occurring within the same time span. These simultaneous events initiated a 'snowball effect' of severe economic depression and a rapid deterioration of the overall public health infrastructure in the country. North Korea's continued isolation and reluctance to release health statistics has left the international community uncertain of the precise extent of the public health devastation. The uncertainty of the situation has been further complicated by disparate accounts of the public health declines. For instance, the North Korean Government has estimated that approximately 220,000 people died due to famine in the 1990s, while the World Health Organization (WHO) claims that this figure is closer to 2 million. In the past few years, the willingness of the North Korean Government to engage the outside world has increased. This is reflected by the growing number of foreign aid organizations or non-governmental organizations (NGOs) that have been permitted entry into the country. NGOs have not only served to improve various aspects of the public health system but also serve as a source of 'on-the-ground' information for the outside world. This role has proven critical not only for public health purposes but for facilitating improved international relations between countries. PMID- 15780322 TI - The Dutch heart health community intervention 'Hartslag Limburg': results of an effect study at organizational level. AB - BACKGROUND: 'Hartslag Limburg', a cardiovascular diseases (CVD) prevention programme, integrates a community strategy and a high-risk strategy to reduce CVD risk behaviours. This article presents the results of the effect evaluation study of the community intervention at the organizational level. Organizational changes were an intermediate goal of the Hartslag Limburg community intervention, as these are assumed to be a prerequisite for changes at the individual level. METHODS: A baseline-post-test control group design was used. The baseline measurement was conducted in 1998 and the post-test measurement in 2001. At baseline, 700 organizations were selected in the Maastricht region, and 577 in a control region. All organizations that were potentially significant agents in health-promoting activities were included. Data on organizational involvement in health-promoting activities were gathered by means of structured questionnaires, and sent to organization representatives by mail. RESULTS: The overall post-test percentage of organizations involved in at least one activity relating to physical activity was higher in the Maastricht region than in the control region. Furthermore, the number of activities per organization involved in activities relating to healthy eating, smoking behaviour or physical activity was higher in the Maastricht region than in the control region at post-test. CONCLUSIONS: This study provided valuable information about organizational involvement in health promoting activities, as well as important information to consider in future research in this area. Due to the limitations of the study, the importance of measuring change at different social levels in community-based programmes, and the scarcity of effect studies of community interventions at the organizational level, further research on this subject is warranted. PMID- 15780323 TI - Approaches to increase physical activity: reviewing the evidence for exercise referral schemes. AB - OBJECTIVE: To review current evidence of effectiveness for exercise-referral schemes. METHODS: Studies were identified from MEDLINE 1966-2002, EMBASE 1980 2002 and CINHAL 1982-2002 and bibliographies of relevant papers. INCLUSION CRITERIA: Interventions providing access to exercise activities and/or facilities, experimental or quasi-experimental studies, studies with a control group, interventions based in a primary care setting, and interventions including an exercise component with measures of physical activity levels. CONCLUSIONS: Exercise-referral schemes appear to increase physical activity levels in certain populations, namely individuals who are not sedentary but already slightly active, older adults and those who are overweight (but not obese). However, increases in the level of physical activity may not be sustained over time. Further studies are required to assess effectiveness in a range of populations and for different activities, and to find strategies to increase long-term adherence. PMID- 15780324 TI - Beliefs, sexual behaviours and preventive practices with respect to HIV/AIDS among commercial sex workers in Daulatdia, Bangladesh. AB - OBJECTIVES: Despite the rising prevalence of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) since 1994 in Bangladesh, the World Bank found the epidemic to be preventable provided vigorous and prompt action is taken. High-risk heterosexual contact, especially among commercial sex workers (CSWs), is a major mode of transmission. Formulation of relevant and effective prevention programmes for HIV/AIDS requires better understanding of the knowledge, attitudes, behaviours and practices in the high-risk groups. STUDY DESIGN AND METHODS: A cross-sectional survey comprising face-to-face interviews using a structured questionnaire with items on knowledge, beliefs, condom use and other sexually transmitted diseases (STDs). SETTINGS: In total, 300 CSWs were interviewed between July and October 2000 in Daulatdia brothel. Daulatdia is one of the largest river ports in Bangladesh. RESULTS: Although most CSWs had heard of AIDS, correct knowledge of transmission and symptoms was lacking. HIV/AIDS was viewed as a remote threat, over-ridden by immediate economic and survival concerns. Although the majority of CSWs knew that condoms afforded protection against STDs/AIDS, only one-third of sex acts on the last day of work were protected through condom use. CSWs who were married, had been a CSW for less than 5 years, were with a new client, or had two or more clients in last working day reported significantly higher condom use. Client dissatisfaction was the major reason for not using condoms. Many did not obtain treatment for STDs in a timely fashion, if at all. CONCLUSIONS: Bangladesh needs a comprehensive HIV programme that combines clinical and screening measures with behaviour change and communication interventions, along with change in social norms and attention to the rights of CSWs in order to avert a widespread epidemic. PMID- 15780325 TI - Condom-promotion programme among slum-dwellers in Chandigarh, India. AB - This study describes high-risk behaviour and condom usage in inhabitants of an urban slum in the Union Territory of Chandigarh, which has a population of about 20,000. A cohort of 375 participants was followed-up before and after an intervention. Half of the married but only one-eighth of the unmarried slum dwellers were using condoms regularly. One-quarter of the study subjects practised high-risk behaviour and this was higher among unmarried (44.88%) than married participants (18.7%). An intervention involving education on condom use and provision of free condoms increased the condom usage rate from 31.73% in the pre-intervention phase to 60% in the postintervention phase. PMID- 15780326 TI - The prevalence of obesity and associated morbidity among 17-year-old Israeli conscripts. AB - BACKGROUND: There has been a significant increase in the prevalence of obesity among children and adolescents over the last few decades. Obesity is associated with significant psychosocial and physical morbidity. OBJECTIVE: The aim of this study was to define the prevalence of obesity and associated morbidity, including type 2 diabetes and hypertension, among 17-year-old adolescents. A comparison between the morbidity patterns of females and males was also performed. DESIGN: All 17-year-old Israeli nationals are obliged by law to present at the Israel Defence Forces recruiting office for a medical examination, with the exception of orthodox religious and Arabic adolescents. Height and weight are measured and classified, and a trained specialist evaluates those with suspected associated diseases. The level of education is also recorded. RESULTS: Our survey included 76,732 adolescents, 32,402(42.2%) females and 44,330(57.8%) males. The prevalence of obesity and morbid obesity among 17-year-old Israeli conscripts was 4.1% in males and 3.3% in females. The prevalence of borderline overweight was 12.4% in males and 11.4% in females. The prevalence of hypertension and type 2 diabetes was significantly higher among conscripts with BMI>30 kg/m2 in both genders. There was also a significantly higher prevalence of hypertension and type 2 diabetes in males compared with females(P<0.001, 0.015). A significantly higher prevalence of conscripts without high-school education(<10 years of education) was found among females with BMI>30 kg/m2. There was no significant difference among the male conscripts. CONCLUSIONS: This study found a relatively low prevalence of obesity among 17-year-old Israeli conscripts, but an alarmingly high prevalence of borderline overweight. Obesity was correlated with a higher prevalence of hypertension and type 2 diabetes, and a lower level of education. There was a significant difference between genders in the prevalence of hypertension and type 2 diabetes, and this should be further investigated. These data call for the creation of new education programmes on the prevention of overweight among children and adolescents. PMID- 15780327 TI - Smoking habits and attitudes towards smoking among Estonian physicians. AB - OBJECTIVES: This study examined the smoking habits and attitudes towards smoking among Estonian physicians. STUDY DESIGN AND METHODS: Cross-sectional data for 2668 physicians were gathered by a self-administered postal survey. RESULTS: The current smoking prevalence was 24.9% for male physicians and 10.8% for female physicians. The percentages of ex-smokers were 32.9 and 16.8%, respectively. Smoking prevalence among physicians was below the levels reported for the highest educational bracket of the total population in Estonia. Non-smoking physicians had more unfavourable views towards smoking than those who smoked. The majority of physicians were aware of the association between smoking and various diseases, with significant differences between smokers and non-smokers. Non-smoking physicians were more active in asking patients about smoking habits than those who smoked. Most Estonian physicians, especially those who smoked, failed to perceive themselves as positive role models. CONCLUSIONS: This study found a lower prevalence of smoking among physicians compared with the general population, and demonstrated the impact of personal smoking on physicians' attitudes towards smoking. The results provide an important challenge to medical education in Estonia. PMID- 15780328 TI - Standardizing questionnaire items for the assessment of waterpipe tobacco use in epidemiological studies. AB - Lessons from surveillance of the smoking epidemic show that the lack of adequate attention to standardizing measures and instruments for epidemiological studies has negatively influenced our ability to assess spatial and secular trends in smoking worldwide. Waterpipe smoking, another hazardous form of tobacco use, is gaining popularity worldwide, with societies in the Eastern Mediterranean region (EMR) being most affected. Several research groups are currently investigating the epidemiology of waterpipe smoking among various populations in the EMR. Initial evidence shows that in contrast to usage patterns observed in adult cigarette smoking, waterpipe smoking is characterized mainly by intermittent and social use. As such, many measures that have been traditionally used for the study of usage patterns and dependence among adult cigarette smokers are uninformative for waterpipe smoking. Thus, the need to develop standardized measures and terminology for assessment of the epidemiology of waterpipe smoking in different populations is of paramount importance. As the monitoring of waterpipe smoking is in its infancy, the development of consensus measures should facilitate the initiation of effective surveillance that can guide public health response to this emerging epidemic. PMID- 15780329 TI - The trend of pulmonary tuberculosis in patients seen at DOTS clinics in the Federal Capital Territory, Abuja, Nigeria. AB - We evaluated the pattern of sputum smear positivity and assessed the effects of directly observed treatment short course (DOTS) among tuberculosis (TB) patients at the DOTS clinics in the Federal Capital Territory (FCT), Abuja. In total, 1391 patients were seen at six microscopy and treatment centres across the FCT between January and December 2003. Their sputa were screened microscopically for the presence of acid-fast bacilli (AFB) using the Ziehl-Neelsen staining technique. In total, 296 (21.3%) patients were smear positive; 201 (67.9%) were new cases and 95 (32.1%) were follow-up cases. The highest incidence of sputum smear positivity (24.8%) was found in those aged 21-30 years and the lowest incidence (6%) was found in those aged 71 years and above. No incidence of smear positivity was recorded in children aged 0-10 years. In total, 160 of the men screened were AFB positive (75% new cases, 25% follow-up cases). In comparison, 136 women were AFB positive (59.6% new cases, 40.4% follow-up cases). During the 1-year study period, two deaths were recorded. Men pose a serious threat to public health as most of the follow-up cases result in the tubercle bacilli developing resistance to available anti-TB drugs. This study demonstrated a high prevalence of infectious TB in the population screened, and therefore underlines the need for capacity building through a multisectoral approach in the fight against the disease. Cohort analysis should be the cardinal management strategy in evaluating the effectiveness of TB control through systemic follow-up and reporting of certain indicators in treatment progress and success. PMID- 15780330 TI - Caregivers's knowledge, attitude and practice on childhood malaria and treatment in urban and rural communities in Enugu, south-east Nigeria. AB - OBJECTIVE: To ascertain caregivers's knowledge, attitude and practice regarding malaria and treatment of children in rural and urban communities in Enugu, Nigeria. STUDY DESIGN: This was a cross-sectional study, and caregivers were selected by convenience sampling. METHOD: In total, 224 and 184 respondents were interviewed in paediatric outpatient clinics in rural and urban areas, respectively, using structured questionnaires. RESULTS: Most respondents in rural and urban areas has heard of malaria (99% urban, 74% rural; P < or = 0.05). Both groups were unaware of the particular susceptibility of children and pregnant women to the disease, and respondents preferred to use self-medication (urban 79%, rural 20%). The rural and urban caregivers were quite familiar with and used antimalarial drugs such as chloroquine (urban 23%, rural 15%) and sulphadoxine pyrimethamine (urban 50%, rural 6%), although in inappropriate dosages. They were also aware of preventive measures against malaria such as insecticide-treated nets (urban 32%, rural 56%), but there was low usage in both communities (7 and 2%, respectively). CONCLUSIONS: Both rural and urban communities were aware of malaria as a disease, but practices were grossly inadequate. The provision of affordable healthcare facilities and the use of health education to change knowledge, attitude and practice will help meet the target goals of the 'Roll Back Malaria' campaign in the area. PMID- 15780332 TI - Fine particles and meteorological conditions are associated with lung function in children with asthma living near two power plants. AB - Fine particles are thought to pose a risk to health, especially for vulnerable groups such as children with asthma. These children are also known to be affected by meteorological and seasonal changes. We assessed the association between air pollution and lung function via peak expiratory flow (PEF), controlling for seasonal changes, meteorological conditions and personal physiological, clinical and sociodemographic measurements, in a panel of schoolchildren with asthma living near two power plants in Israel. Two hundred and eighty-five children with confirmed asthma performed PEF tests and completed a respiratory symptoms diary twice a day. Particulate matter <10 microm in diameter (PM10), particulate matter <2.5 microm in diameter (PM2.5) and meteorological conditions were measured at six fixed stations. Data were analysed using time series analysis-generalized linear model and generalized estimating equations. The models were built under the assumption that any health outcome belongs to a multivariate hierarchical system and depends on meteorological, geophysical and sociocultural variables and pollution factors. No significant differences were found in the demographic (age, gender, mean parental education level, parental smoking habits, place of birth and housing density), physiological (body mass index) and clinical factors (illness severity) between the communities participating in the study. A significant direct effect of PM2.5 on the PEF was found in Ashdod (P=0.000). In Sderot, this effect was through an interaction between PM10 and the sequential day of the year (P=0.000). The main conclusion of this study is that children with asthma are at risk from air pollution and geophysical conditions. Policy makers should take these results into consideration when setting thresholds for environmental protection. PMID- 15780331 TI - The use of personal protection measures against malaria in an urban population. PMID- 15780333 TI - Drinking patterns and traffic casualties in Switzerland: matching survey data and police records to design preventive action. AB - OBJECTIVE: To examine the association between drinking patterns and alcohol related traffic casualties. STUDY DESIGN: Data linkage of cross-sectional survey data on alcohol consumption with official traffic casualty records. METHODS: Alcohol consumption measures for usual heavy drinking and risky single occasion drinking were derived for different time segments of the day from a 7-day drinking diary study of 747 current drinkers. Measures were correlated with official records of alcohol-related traffic casualties. RESULTS: There was a high correlation between alcohol-related traffic casualties and the number of risky single occasion drinkers that consumed alcohol outside their homes (r=0.92). On average, about 50% of these drinking occasions were attributed to usual moderate drinkers. The proportion of usual heavy drinkers was lowest in the time segments with the most alcohol-related casualties. CONCLUSION: Preventive countermeasures should be targeted at the general population, enforced particularly during specific periods of the week. PMID- 15780334 TI - Assessment of cardiovascular risk factors and their correlation in 28,000 Hungarians. AB - OBJECTIVES: The aim of this study was to assess the prevalence and inter relations of systolic hypertension, hypercholesterolaemia and hyperglycaemia in more than 28,000 Hungarians. STUDY DESIGN AND METHODS: A retrospective study was designed to collect relevant information from the database of the Screening Institution in Miskolc, Hungary, between 1990 and 1998. Prevalence of the investigated conditions was compared between different groups. RESULTS: The occurrence of any of the investigated cardiovascular risk factors increased the likelihood of the other risk factors, and this relationship was most apparent in more serious categories. A particularly high degree of comorbidity was found between hypertension and hyperglycaemia. CONCLUSION: Since these results were based on a large percentage of the population, this study provides an important message for practising doctors: if they find any of these three risk factors in a patient, it is highly recommended that they should search for the others. PMID- 15780335 TI - Differences in risk of mortality under 1 year of age between rural and urban areas: an ecological study. AB - OBJECTIVE: To investigate differences in risk of categories and causes of death before 1 year of age between rural and urban areas. METHODS: Population-based ecological study using Poisson regression analysis of data from all enumeration districts in Wales. Data included all 243,223 registrable births to women resident in Wales, 809 therapeutic and spontaneous abortions, 1302 stillbirths and 1418 infant deaths occurring between 1993 and 1999. MAIN RESULTS: The relative risk of mortality in rural areas compared with urban areas for all deaths before 1 year of age was 0.89 (95% confidence interval 0.82, 0.98, P=0.02). The risk of mortality in rural areas was significantly lower than in urban areas for all categories of deaths occurring after 7 days of life. The relative risk of death due to infection was significantly lower in rural areas compared with urban areas (P=0.04), with similar results for deaths due to sudden infant death syndrome (P=0.03). After adjusting for social deprivation, there were no significant differences in the risk of death between rural and urban areas. CONCLUSIONS: While there were significant differences in crude risk between rural and urban enumeration districts for some causes and age groups before 1 year, after adjusting for social deprivation, these differences were not significant. The lack of significant interaction between rurality and deprivation indicated that the relationship between social deprivation and death before 1 year of age was not significantly different in rural areas compared with urban areas. Collaborative public health programmes to tackle deprivation are necessary in both rural and urban areas. PMID- 15780336 TI - Evaluation of an innovative approach to the Director of Public Health's annual report. AB - BACKGROUND: The aim of this study was to evaluate the process of an innovative 'presentation with discussion' approach to the Director of Public Health's (DPH) annual report for North Derbyshire Health Authority. STUDY DESIGN: Mixed methods survey and interviews. METHODS: Survey methods were used to obtain qualitative and quantitative data. Key stakeholders were sent a postal questionnaire, individuals attending presentations could respond via a printed slip or letter, structured group discussions following presentations were recorded manually, and presenters and producers were interviewed. RESULTS: Thirty-five of 41 (85.4%) key stakeholders returned questionnaires, 18 people responded individually, discussions following 26/30 (86.7%) presentations were documented, and all eight presenters and 11 producers were interviewed. The general response was extremely positive with 25 of 35 (71%) key stakeholders, 100% of presenters and 80% of producers preferring the new format to the previous year's report. People felt that it conveyed the public health message effectively, and appreciated the opportunity to contribute to the recommendations for improving health in North Derbyshire. Many were concerned that the circulation should be wide, and key stakeholders were keen to have a printed reference document to support their work. Presenters enjoyed the process although tailoring the presentations to unfamiliar audiences proved difficult. The production team found the process more efficient and focused, although direct production costs were increased. Presenters and producers were concerned that the process for implementing recommendations for action was not fully considered. CONCLUSIONS: Most people preferred this innovative approach to the traditional annual report, and it is therefore recommended that this format should be used for future DPH annual reports. Key stakeholders still require a reference document, and presenters' briefing notes could be adapted for this purpose. Circulation of the report should be wide, and further consideration of how participants' recommendations for action are implemented is needed. PMID- 15780337 TI - Chronic diseases: an increasing challenge in developing countries. PMID- 15780338 TI - Evaluating a malaria intervention strategy using knowledge, practices and coverage surveys in rural Bolifamba, southwest Cameroon. AB - We evaluated the impact of a malaria intervention in Bolifamba in rural Cameroon. The intervention consisted of educating the community on management of malaria and provision of a dispensary for early diagnosis and treatment. In July 2001, prior to the intervention, a questionnaire was used to obtain information on knowledge of and practices toward childhood malaria of 185 mothers of children aged 0-5 years. The same questionnaire was administered to 120 of the 185 mothers, one-year post-intervention. Clinical and laboratory investigations were carried out on children whose mothers were interviewed. A comparison of pre- and post-intervention data indicated significant changes in (i) the use of appropriate malaria treatment (from 50% to 81.7%); (ii) recognition of splenomegaly as a feature of malaria (from 18.4% to 80.8%); (iii) prevalence of splenomegaly (from 26.5% to 13.3%); (iv) prevalence of fever (from 27.8% to 13.3%); (v) parasite prevalence (from 60.5% to 44.2%) and (vi) severe malaria anaemia (from 2.6% to 0.0%). These findings revealed that proper education of villagers, particularly mothers, on malaria and the presence of health facilities, where treatment is readily available at affordable cost, close to villages, are important strategies that would reduce malaria morbidity and mortality significantly. PMID- 15780339 TI - Curdlan sulphate in human severe/cerebral Plasmodium falciparum malaria. AB - Preclinical studies have shown that curdlan sulphate (CRDS), a sulphated 1-->3 beta-D glucan, inhibits Plasmodium falciparum in vitro and down-modulates the immune response. A direct, non-specific effect on cytoadherence and rosetting may be predicted, as has been described with other sulphated polysaccharides, e.g. heparin. The anticoagulant effect of CRDS is 10-fold lower than heparin. Curdlan sulphate has, therefore, emerged as a candidate for adjunct medication in the treatment of severe/cerebral malaria. Two clinical studies were conducted using CRDS as adjunct medication to conventional therapy (artesunate) in patients with severe and severe/cerebral malaria. Both studies were double-blind and placebo controlled to evaluate the efficacy and safety of the combination. Curdlan sulphate appeared to reduce the severity of the disease process, e.g. fever clearance time was shortened. Due to the small number of patients, there was no difference in mortality. The two treatment arms in both studies showed similar results for all laboratory parameters. The only adverse event recorded during CRDS treatment was an increase in activated partial thromboplastin time. This can be monitored easily. It seems that the patients who may benefit most are severe/cerebral cases with no organ damage on admission. PMID- 15780340 TI - Why has the dihydrofolate reductase 164 mutation not consistently been found in Africa yet? AB - Resistance to the antifolate sulfadoxine-pyrimethamine (SP), the current mass treatment antimalarial drug, is associated with selection of point mutations in dihydrofolate reductase and dihydropteroate synthase. Among these mutations, the leucine 164 dihydrofolate reductase mutation (Leu-164) is associated with higher levels of SP resistance; this mutation is also associated with a decrease in the efficacy of chlorproguanil/dapsone, a newly developed antifolate antimalarial drug. Leu-164 has been detected in Southeast Asia and South America, regions where SP is no longer effective. Surprisingly, this mutation has not yet been detected in Africa, using the standard protocol based on PCR-RFLP, despite high SP resistance. In this paper, we discuss briefly the reasons why Leu-164 has not yet been selected in Africa and we propose a means that may slow down the selection of this mutation. PMID- 15780341 TI - Malaria vectors and urbanization in the equatorial forest region of south Cameroon. AB - Entomological surveys were carried out in the town of Mbalmayo and in the nearby rural village of Olama, within the equatorial forest zone of Cameroon. Mosquitoes were captured when landing on human volunteers and by pyrethrum spray catches. Malaria vectors captured were Anopheles gambiae Giles (M and S forms) and A. moucheti Evans in both areas, together with A. funestus Giles in Mbalmayo. One A. marshallii (Theobald) specimen infected by Plasmodium falciparum was found in Olama. Anopheles moucheti was the most abundant anopheline species caught in Olama, while A. gambiae was the most abundant in Mbalmayo. All these vectors were highly anthropophilic as indicated by the fact that only 5 of 201 blood meals analysed had been taken from non-human hosts. Plasmodium falciparum was the only malaria parasite species found in Mbalmayo, while P. malariae was also found in Olama. The annual entomological inoculation rate was estimated at 129 infective bites/person/year in Mbalmayo and 322 in Olama. Comparison with data published in 1955 from Mbalmayo, before expansion of the town, showed the impact of urbanization on the composition of the vector system and malaria transmission dynamics. Such changes should be considered when implementing sustainable control measures. PMID- 15780342 TI - Factors affecting delays in tuberculosis diagnosis in rural China: a case study in four counties in Shandong Province. AB - Improving case detection is an urgent and serious challenge for tuberculosis (TB) control in China. We investigated the extent to which TB patients delayed seeking TB care and health services delayed reaching a diagnosis, and socio-economic factors associated with the delays. Standard questionnaires were administered to 190 new smear-positive TB patients who had completed treatment at TB dispensaries in four counties of Shandong Province in 2001. Multivariate analysis using Cox Regression showed that old age, lack of education and distance from home to a township health centre were significantly associated with delay in seeking care from service providers. In examining the delay between first contact with a service provider and diagnosis, we found that women experienced longer delays than men, and that the higher the level of facility patients first visited, the less time was needed to achieve a diagnosis. These two factors were statistically significant in multivariate Cox Regression analysis. We concluded that the elderly, the less educated, women, and those living far from health facilities face the longest delays in reaching TB services and achieving diagnosis. PMID- 15780343 TI - PPD induced in vitro interferon gamma production is not a reliable correlate of protection against Mycobacterium tuberculosis. AB - Correlates of protection against tuberculosis are crucial for the evaluation of new vaccine candidates and for the demonstration of their potential efficacy. Such correlates can be proposed on the basis of animal models. In this study, we hypothesized that protection against tuberculosis (TB) induced by bacillus Calmette-Guerin (BCG) correlates with in vitro TB antigen-specific IFN-gamma production. BCG vaccination, known to provide effective protection against TB in animals, was used to investigate the use of in vitro IFN-gamma production as a marker of BCG-induced protection against TB. Our results show that BCG vaccination does provide substantial protection against challenge with Mycobacterium tuberculosis. However, despite previous compelling evidence that Th1 type immune responses are essential for TB immunity, the magnitude of in vitro purified protein derivative (PPD)-specific IFN-gamma production assessed during the course of TB infection did not correlate with protection. This emphasizes the need to identify further correlates of protection, in addition to IFN-gamma, to be used as markers of protective immunity against M. tuberculosis and/or to identify M. tuberculosis antigens inducing IFN-gamma that correlate with protective immunity. PMID- 15780344 TI - Field evaluation of a recombinant glutathione S-transferase-based pyrethroid quantification assay. AB - A recombinant glutathione S-transferase (GST)-based pyrethroid quantification assay was field-tested in Ifakara, Tanzania. Initial laboratory tests suggested that all reagents used in the assay should be sufficiently stable for field use, provided that domestic refrigeration facilities were available. Insecticide impregnated bednets were collected from a region where a social marketing programme was in progress. A total of 100 bednets were collected and the assay plus standard HPLC analysis was performed on the residues extracted from four replicate areas of each net. Insecticide residue estimations for assays performed on white and pale green bednet samples were accurate when compared with residue analysis by HPLC. However, for dark green or blue bednets, there was no correlation between the GST-based assay and HPLC pyrethroid quantification results. The assay failure with the dark coloured nets was caused by the extraction of the dyes along with the insecticide, which subsequently interfered with the GST assay. When the same samples were analysed by HPLC, the dyes were separated from the insecticide by reverse phase column chromatography and hence did not affect the results. PMID- 15780345 TI - Trypanosoma cruzi infection in wild mammals of the National Park 'Serra da Capivara' and its surroundings (Piaui, Brazil), an area endemic for Chagas disease. AB - We studied the prevalence of Trypanosoma cruzi infection among eight species of wild small mammals (n=289) in an area where human cases of infection/disease have occurred. Dogs (n=52) and goats (n=56) were also surveyed. The study was carried out inside a biological reserve, the National Park 'Serra da Capivara' and its surroundings in Piaui State, Brazil. The marsupial Didelphis albiventris and the caviomorph rodent Trichomys apereoides were found to be the most important reservoirs in the study area. Trichomys apereoides was the most abundant species (80%) and D. albiventris the most frequently infected (61%). Both T. cruzi I and T. cruzi II genotypes were isolated from these species. One specimen of Tr. apereoides displayed a mixed T. cruzi I/zymodeme 3 infection. Serum prevalence among dogs suggests that they may be involved in the maintenance of the parasite in the peridomestic environment, in contrast to goats, which are not apparently of any epidemiological importance. The distinct distribution and patterns of infection observed in the study areas suggest that even in the same biome, epidemiological studies or determination of control measures must take into account ecological peculiarities. PMID- 15780346 TI - Distribution of Plasmodium falciparum genotypes in clinically mild and severe malaria cases in Orissa, India. AB - A cross-sectional study was conducted in a malaria hyperendemic state of India to ascertain the distribution of Plasmodium falciparum genotypes in patients with mild (n=40) and severe (n=35) malaria. PCR and nested PCR were used to determine the glutamate-rich protein (GLURP), merozoite surface proteins 1 and 2 (MSP1 and MSP2) and knob-associated histidine-rich protein (KAHRP) for characterization of the parasite. The results indicate that (i) the 200bp allele of the MAD20 family of MSP1 and the 550bp allele of the 3D7 family of MSP2 show over-representation in severe malaria cases; (ii) the multiplicity of infection with respect to MSP2 alleles is significantly higher (P<0.001) in severe cases than in mild cases; and (iii) comparison with the findings of other studies leads to the conclusion that the distribution of P. falciparum genotypes between different clinical groups differs geographically. PMID- 15780348 TI - Renal cryoablation: study with care--proceed with caution. PMID- 15780347 TI - Renal cryotherapy: pro. PMID- 15780349 TI - Minimally invasive surgery for pelviureteral junction obstruction in adults: a critical review of the options. PMID- 15780350 TI - Obesity and prostate cancer. PMID- 15780351 TI - Bacterial colonization on intraluminal surface of urethral catheter. AB - OBJECTIVES: To determine the relationship between catheter-colonizing bacteria and urinary planktonic bacteria. The significance of catheter-colonizing bacteria for the pathogenesis of urinary tract infection remains unclear, although they are involved in the development of catheter-associated urinary tract infection refractory to antimicrobial chemotherapy. METHODS: The subjects were 86 inpatients with urethral catheters. Urinary culture and catheter culture were performed simultaneously. The intraluminal catheter surface was swabbed to prepare a suspension. The suspension and a urine specimen from each patient were quantitatively cultured with the dip slide method and the microorganisms identified. RESULTS: The catheters had been mainly placed to monitor urine output after urologic surgery, and their median indwelling period was 3.0 days (range 1 to 35). The overall positive rate of catheter culture was significantly greater than that of urine culture (53.5% and 30.2%, respectively, P <0.01), even in patients without a recent antibacterial agent history. The difference was observed at day 2 (60% and 13.3%, catheter versus urine culture, respectively, P = 0.011) and days 3 to 6 (52.4% and 14.3%, respectively, P = 0.010) of the indwelling period, but was indistinguishable at day 14 and thereafter. The percentage of patients who had the same bacterial species isolated from both specimens increased in a time-dependent manner. CONCLUSIONS: The present results revealed considerable differences in the positive rates and bacterial species isolated from urine and catheter cultures, indicating that not all species of bacteria colonizing the intraluminal surface of the urethral catheter are detected as urinary bacteria. These results also suggest that bacterial colonization on the intraluminal catheter surface can precede the emergence of bacteriuria. PMID- 15780352 TI - Diagnosis of prostatic inflammation: efficacy of needle biopsies versus tissue blocks. AB - OBJECTIVES: To assess the variation of inflammation found on single needle biopsies, we compared our results using multiple needle biopsies and surgical samples taken from predetermined areas of prostates removed from patients with prostate cancer. METHODS: Biopsies and tissue blocks were taken from 25 prostates that had been removed at prostate cancer surgery. These samples were taken from the contralateral side from the cancer whenever possible. The biopsies were taken from the apex, middle, and base, and blocks were taken from the peripheral and transition zones of the middle region. The biopsies were evaluated for density of inflammatory cells and their histologic location (glandular vs. periglandular vs. stromal). RESULTS: Inflammation was generally periglandular, with multifocal and diffuse patterns. The severity of the inflammation was quite variable. The overall inflammatory cell counts did not differ among the regions except for lower levels at the base. Correlation coefficients demonstrated significant correlations of the middle region biopsies with the peripheral zone and transition zone blocks (P = 0.02 and P = 0.016); the apical biopsies also correlated with the transition zone blocks. CONCLUSIONS: Our results suggest that individual, transrectal needle biopsies taken consistently from the peripheral zone or transition zone of the apex or middle region of prostates from patients with incidental prostatitis may serve as well as multiple biopsies for evaluating the degree of inflammation. PMID- 15780353 TI - Postoperative acute confusional state in typical urologic population: incidence, risk factors, and strategies for prevention. AB - OBJECTIVES: To determine the incidence of, and predictors for, the acute confusional state (ACS) in older patients after urologic surgery. ACS is among the most common complications after surgery in older patients. It is associated with increased postoperative morbidity, longer hospital stays, and greater mortality. Agitation caused by ACS might have deleterious consequences in a large proportion of older patients, especially after urologic surgery. Only a few studies, of highly selected urologic procedures, have been reported, and all showed an astonishingly low percentage of patients with this distressing condition. METHODS: We examined 100 consecutive, older patients (age 60 years or older), prospectively, before and after urologic surgery, to determine both the incidence of, and the predictors for, ACS. RESULTS: Only 7 of the 100 patients developed postoperative ACS. The risk factors identified were preoperative cognitive deficits, pre-existing depression, impaired vision, and the operative time. CONCLUSIONS: These results suggest that postoperative ACS is relatively rare after urologic surgical procedures; however, patients who are likely to develop ACS can be identified, prompting consideration for prophylactic antidelirium care. PMID- 15780354 TI - Percutaneous nephrolithotomy with holmium: Yttrium-aluminum-garnet laser and fiber guider--report of 349 cases. AB - OBJECTIVES: To assess the efficacy and safety of holmium:yttrium-aluminum-garnet (YAG) laser lithotripsy for percutaneous nephrolithotomy in a cohort of 349 consecutive procedures. METHODS: We retrospectively reviewed the charts of 334 patients who had undergone 349 percutaneous nephrolithotomy (PCNL) procedures conducted at our hospital from January 2001 to March 2004 in which the holmium:YAG laser with a self-made fiber guider was used for lithotripsy. RESULTS: The average patient age was 54.1 years, and the average stone size was 3.3 +/- 1.8 cm. The average operative time was 99 +/- 38 minutes, and the average postoperative hospital stay was 4.0 +/- 2.3 days. The overall stone-free rate was 83.7%. The postoperative urinary tract infection rate was 7.2%. The postoperative transfusion rate was 2.0%. The holmium:YAG laser was effective against all kinds of stones, but sometimes it was time consuming to use the holmium:YAG laser to disintegrate a very large stone. Thirteen PCNLs (3.7%) to treat large complete staghorn stones were performed in combination with a pneumatic lithotriptor. Of the 349 procedures, 152 (43.6%) had bloodless tracts afterward, and all 152 procedures had been performed with the nephrostomy tube-free modification. CONCLUSIONS: The holmium:YAG laser is an effective and safe lithotriptor for most percutaneous stone surgery. However, in patients with a very large stone burden, the combination of this technology with another, more powerful, intracorporeal lithotriptor may be necessary. PMID- 15780355 TI - Is 24-hour urinary calcium a surrogate marker for dietary calcium intake? AB - OBJECTIVES: To determine whether the 24-hour urinary excretion of calcium was a reliable surrogate marker for dietary calcium intake. Although dietary calcium intake has been negatively correlated with the risk of recurrent calcium-based stones, detailed dietary histories are not routinely evaluated in most patients with recurrent stone formation. METHODS: The dietary records and corresponding 24 hour urine collections of 68 randomly selected women with a history of calcium based renal stones and two or more outpatient clinic visits were studied. Subjects were excluded if they had conditions or took medications affecting calcium absorption or excretion. Multivariate regression analysis was performed on the most recent set of data per patient with 24-hour urinary calcium as the dependent variable. Independent variables included age, weight, and dietary calcium, sodium, potassium, magnesium, fiber, and animal protein. Regression analysis was performed on the differences between the first and last visits for dietary and urinary data. Using tertiles, the positive predictive value of 24 hour urinary calcium for the respective dietary intake tertiles was calculated. RESULTS: The regression model on absolute values showed all dietary parameters to have squared partial correlation coefficients of less than 0.3 (P = 0.015, R2 = 0.264). In the second regression analysis, the model did not significantly explain the variance (P = 0.656). The positive predictive value of a mean 24-hour urinary calcium level less than 3.75 mmol/L for calcium intake less than 585 mg/day was 45%. CONCLUSIONS: The results of the present study revealed that the 24-hour urinary calcium cannot be used as a surrogate marker for dietary intake of calcium. A detailed dietary history is needed for all women with recurrent stone formation for proper assessment and potential modification of calcium intake to reduce recurrences. PMID- 15780356 TI - Improved hemostasis during laparoscopic partial nephrectomy using gelatin matrix thrombin sealant. AB - OBJECTIVES: To evaluate whether using a biologic hemostatic sealant facilitates hemostasis during laparoscopic partial nephrectomy. Secure and durable parenchymal hemostasis is a critical requirement during laparoscopic partial nephrectomy. METHODS: Since September 1999, laparoscopic partial nephrectomy has been performed in more than 300 patients by a single surgeon, duplicating open surgical principles. Recently, from patient 225 onward, we modified our technique by incorporating topical application of a gelatin matrix thrombin sealant (FloSeal) to cover the partial nephrectomy bed before sutured renorrhaphy over a Surgicel bolster. The impact of FloSeal on reducing hemorrhagic complications was evaluated by comparing two sequential groups of patients: group 1 consisted of 68 patients in whom FloSeal was not used (patients 156 to 224) and group 2 consisted of 63 patients in whom it was used (patients 225 to 288). RESULTS: Groups 1 (no FloSeal) and 2 (FloSeal) were comparable in tumor size, number of central tumors, and performance of pelvicaliceal suture repair (84% versus 92%; P = 0.16). Intraoperative variables were also comparable in terms of mean warm ischemia time (36.1 versus 37.2 minutes; P = 0.55), blood loss (150 versus 106 mL; P = 0.36), operative time, and hospital stay. However, the FloSeal group had significantly fewer overall complications (37% versus 16%; P = 0.008) and tended toward a lower rate of hemorrhagic complications (12% versus 3%), although this did not achieve statistical significance (P = 0.08). CONCLUSIONS: The results of this study have shown that adjunctive use of gelatin matrix thrombin sealant substantially enhances parenchymal hemostasis and has decreased our procedural and hemorrhagic complications to levels comparable with contemporary open partial nephrectomy series. This gelatin matrix-thrombin tissue sealant is now a routine part of laparoscopic partial nephrectomy at our institution. PMID- 15780357 TI - Comparison of nephron-sparing surgery in central versus peripheral renal tumors. AB - OBJECTIVES: To determine the feasibility of nephron-sparing surgery (NSS) in patients with centrally located tumors. METHODS: A retrospective cohort study of 118 patients who underwent NSS between 1993 and 2002 (35 patients with centrally located tumors and 83 with peripherally located tumors) was performed. Kaplan Meier curves were constructed to evaluate freedom from local recurrence and disease-specific survival in patients with conventional histologic subtype tumors. The Wilcoxon test was used to compare the curves (two-tailed P < or =0.05 was considered to be statistically significant). RESULTS: Intraoperatively, in patients with centrally located tumors, the need to close the collecting system (P = 0.035) and for blood transfusions (P = 0.033) was greater. Two perioperative deaths occurred in patients with peripherally located tumors. Two patients with centrally located tumors subsequently underwent nephrectomy. Of the patients with centrally located tumors, 1 patient had a positive margin, 2 patients had local recurrence, and 1 patient developed metastasis. No positive surgical margins or local recurrence was found in patients with peripherally located tumors, although 4 patients developed distant metastasis. Kaplan-Meier curves for patients with conventional histologic subtype tumors demonstrated a statistically significant difference for local recurrence (P = 0.04), but not for survival (P = 0.71). The mean follow-up time was 38.8 and 43.8 months for patients with centrally located and peripherally located tumors, respectively. CONCLUSIONS: NSS can be used to postpone, or eliminate the need for, nephrectomy in 94.3% of patients with centrally located tumors and can achieve oncologic disease control similar to that for exophytic lesions. These data indicate that NSS should be considered even for patients with centrally located tumors, taking into account that performing such surgery is a challenging task. PMID- 15780358 TI - Laparoscopic heminephrectomy for tumor. AB - OBJECTIVES: To present our experience with laparoscopic heminephrectomy. We defined heminephrectomy as excision of 30% or more of the renal parenchyma. Laparoscopic partial nephrectomy has typically been performed for smaller, exophytic tumors. With growing experience, we have performed more extensive resections for larger tumors, when indicated. METHODS: Since August 1999, we have performed laparoscopic partial nephrectomy in more than 300 patients. Of these, 41 patients were deemed to have undergone laparoscopic heminephrectomy (group 1). A contemporary group of 41 consecutive patients who underwent laparoscopic partial nephrectomy (less than 30% resection) were retrospectively identified for comparison (group 2). RESULTS: The preoperative patient demographics were similar, except for a greater body mass index (P = 0.02) in group 1. Group 1 had larger tumors (3.7 cm versus 2.3 cm, P <0.001) that were more commonly centrally located (41% versus 9.8%, P = 0.001) and more deeply infiltrating (P <0.001) compared with group 2. Group 1 underwent larger parenchymal resections (P <0.001) and routine pelvicaliceal suture repair (P = 0.002). The warm ischemia time was longer in group 1 (39 versus 33 minutes, P = 0.02); however, the amount of blood loss (150 versus 100 mL, P = 0.28) and total operative time (220 versus 190 minutes, P = 0.09) were comparable between the groups. The analgesic requirements, hospital stay, overall complications, and postoperative serum creatinine were comparable between the groups. Histopathologic examination showed that all 82 surgical margins were negative. CONCLUSIONS: Laparoscopic heminephrectomy is an advanced procedure that can be performed efficaciously with equivalent outcomes to those of less substantial resections. To our knowledge, experience with laparoscopic heminephrectomy has not been previously described. PMID- 15780359 TI - Tension-free vaginal tape in the elderly: is it a safe procedure? AB - OBJECTIVES: To analyze the safety and efficacy of tension-free vaginal tape (TVT) surgery in elderly versus younger women with stress urinary incontinence (SUI). METHODS: A total of 157 consecutive elderly women, aged 70 years and older, and 303 younger women (mean age, 74.8 and 57.2 years, respectively) who underwent TVT for urodynamically-confirmed SUI were prospectively enrolled. Concomitant genitourinary prolapse repair was performed in 84% of the elderly and 67% of the younger women. The main outcome measures were perioperative morbidity, postoperative SUI, persistent or de novo urge incontinence, and voiding dysfunction. RESULTS: The incidence of TVT-related morbidity was similar in both groups, except for significantly fewer cases of bladder perforation among elderly women (1.3% versus 4.9%, P <0.05). However, some age-related morbidity was noted among the elderly women: 2 cases of pulmonary embolism, 2 cases of cardiac arrhythmia, 1 case of severe pneumonia, and 1 case of deep vein thrombosis. The outcome analysis was restricted to 123 elderly and 208 younger women with follow up of at least 12 months (mean, 30 +/- 12 months; range, 12 to 67 months). The incidence of persistent postoperative SUI and persistent urge incontinence was similar in both age groups. However, de novo urge incontinence was significantly more common among elderly women (18% versus 4%, P <0.05). Two elderly and three younger women had postoperative pressure-flow studies suggestive of bladder outlet obstruction. CONCLUSIONS: Tension-free vaginal tape surgery in elderly women is associated with good outcome results; however, the risk of postoperative de novo urge incontinence, as well as age-related morbidity, is increased. PMID- 15780360 TI - Perioperative complications of surgery for genital prolapse: does concomitant anti-incontinence surgery increase complications? AB - OBJECTIVES: To establish whether the short-term risks of prolapse surgery are increased by a concomitant anti-incontinence procedure. METHODS: Using data from the past 4 years of the National Hospital Discharge Survey, we identified women admitted for surgery for pelvic organ prolapse (specifically, suspension of the vagina or obliteration of the cul-de-sac, with or without concomitant hysterectomy or vaginal repairs). We identified women diagnosed with complications (including infections, bleeding complications, surgical injuries, pulmonary complications, and cardiovascular complications). We used multiple logistic regression analysis to compare women with and without complications with respect to concurrent continence procedures, controlling for demographic characteristics, concurrent procedures, and medical comorbidity (using the Charlson index). RESULTS: Of the 1931 women who underwent prolapse surgery, concomitant anti-incontinence procedures were performed in 514 (26.6%). Complications were reported in 288 patients (14.9%) and were associated with medical comorbidity (odds ratio 11.2) and concomitant hysterectomy (odds ratio 1.5). Concomitant surgery for incontinence was not associated with an increased risk of complications. CONCLUSIONS: These data suggest that medical comorbidity is strongly associated with complications after surgery for pelvic organ prolapse. A concomitant anti-incontinence procedure did not significantly increase the immediate morbidity of prolapse surgery. PMID- 15780361 TI - Two-surgeon versus single-surgeon radical cystectomy and urinary diversion: impact on patient outcomes and costs. AB - OBJECTIVES: To examine the difference in charges and outcomes between patients who underwent radical cystectomy and urinary diversion by a team of two surgeons versus a single surgeon. METHODS: A total of 63 patients with bladder cancer who underwent the procedures were retrospectively analyzed. Two surgeons sequentially performed the cystectomy and ileal conduit (IC, n = 17) or neobladder (NBL, n = 18) or a single surgeon performed both the cystectomy and IC (n = 21) or NBL (n = 7). Procedure-related charges, hospital charges, operating room time, length of stay, and complications were compared between the two groups. RESULTS: For the IC patients, the two-surgeon team had 60% greater mean surgeon charges (P <0.0001), 23% lower mean anesthesia charges (P <0.0001), 121 minutes shorter operating room time (P = 0.001), and 30% lower operating room charges (P = 0.001). For the NBL patients, the two-surgeon team had 32% greater surgeon charges (P <0.0001), 22% lower anesthesia charges (P = 0.003), 149 minutes shorter operating room time (P <0.0001), and 41% less operating room charges (P <0.0001). No differences were found in total hospital charges. The NBL patients who underwent surgery by two surgeons had a longer length of stay (P = 0.008). No differences were found in complications between the groups. CONCLUSIONS: For IC patients, our data showed no differences in the average overall charges, whether a two-surgeon team or a single surgeon performed the procedure. Additional reductions in hospital charges could offset the greater physician charges in the NBL patients and allow full realization of the benefit from the shorter operative time with the two-surgeon team. PMID- 15780362 TI - Accuracy of two noninvasive methods of diagnosing bladder outlet obstruction using ultrasonography: intravesical prostatic protrusion and velocity-flow video urodynamics. AB - OBJECTIVES: We assessed the accuracy of two noninvasive, ultrasound methods of diagnosing bladder outlet obstruction (BOO). The potential for the combined methods to enhance the accuracy of diagnosis was also assessed. METHODS: We evaluated 30 male outpatients using two recently developed diagnostic methods, transabdominal ultrasound grading of intravesical prostatic protrusion (IPP) and Doppler ultrasound urodynamics (Doppler UDS), as well as conventional pressure flow studies according to the research protocol. IPP has been reported to be a useful anatomic parameter for the assessment of BOO. Doppler UDS uses transperineal ultrasonography to measure the velocity flow of urine at the prostatic and sphincteric urethras. We recruited 168 outpatients; however, of the 57 patients who fulfilled all the research criteria, only 30 were able to undergo conventional pressure flow study analysis. The results of all three diagnostic methods were analyzed statistically for reliability and relationship in the 30 patients with a BOO index from the conventional pressure flow study. RESULTS: IPP grading correlated well with the BOO index (Spearman's rho 0.624), as did the Doppler UDS parameter (Spearman's rho 0.736). The combination of IPP grading and Doppler UDS showed good sensitivity and specificity. CONCLUSIONS: We confirmed the accuracy of the two novel methods. The combination of the two methods may be a novel standard in the diagnosis of BOO in male patients. PMID- 15780363 TI - Comprehensive study of bladder neck contracture after transurethral resection of prostate. AB - OBJECTIVES: To test the validity of transurethral resection of the prostate (TURP) plus transurethral incision (TUI) of bladder neck as an alternative to TUI of the prostate, a nonrandomized and retrospective study was done to review comprehensively the incidence of, severity of, and risk factors for bladder neck contracture (BNC) in patients with benign prostatic hyperplasia who underwent transurethral surgery. METHODS: The evaluation parameters included age, prostate specific antigen level, urinalysis and uroflowmetry findings, voided volume, presence of vesical stones and urinary retention, surgical type, adenoma weight, and perioperative morbidities. RESULTS: Of the 1470 patients studied, 1135 (77.2%) were eligible for analysis. At a mean follow-up of 37.9 months, 110 patients (9.7%) had developed BNC. The adenoma weight, blood transfusion, and postoperative maximal and mean flow rate in patients with BNC were significantly less than in patients without BNC. The incidence of BNC in the TURP group was greater than that in the TURP plus TUI group (12.3% versus 6%, P = 0.000). BNC was completely prevented using TURP plus TUI if the adenoma weight was greater than 30 g. However, in the TURP group, 4% of patients developed BNC even with an adenoma weight greater than 50 g. Multivariate analyses showed that adenoma weight and surgical type were significant risk factors for BNC. Patients with associated vesical stones were less prone to develop BNC. CONCLUSIONS: TURP plus TUI provides a pathologic diagnosis, and with minimal morbidity, it can be an alternative to TUI of the prostate in selected patients. Personal diathesis may play a role in the pathogenesis of BNC. PMID- 15780365 TI - Analysis of acute toxicity with use of transabdominal ultrasonography for prostate positioning during intensity-modulated radiotherapy. AB - OBJECTIVES: To analyze the effects of the B-mode ultrasound acquisition and targeting (BAT) system for positioning of patients with prostate cancer receiving intensity-modulated radiotherapy on acute gastrointestinal (GI) and genitourinary (GU) toxicity. METHODS: The records of 50 consecutive patients treated using the BAT system were reviewed. Additionally, a comparison (no-BAT) group (ie, a group without a BAT study) treated in a similar manner was identified. The no-BAT group consisted of 49 patients treated immediately before the BAT group. For the two groups, the target definitions and dose prescriptions were identical, the treatment plan acceptance criteria were identical, and intensity-modulated radiotherapy was used for all patients. The daily BAT movements were charted in each of the three principal directions. Acute toxicity was scored for all patients according to the Radiation Therapy Oncology Group GI and GU acute toxicity scales. RESULTS: The GU toxicity rates for the BAT versus no-BAT groups were grade 0 in 20% versus 14%; grade 1 in 38% versus 47%; grade 2 in 38% versus 39%; and grade 3 in 4% versus 0%, respectively (P = 0.284). The corresponding GI toxicity rates were grade 0 in 42% versus 27%; grade 1 in 28% versus 29%; and grade 2 in 30% versus 45% (P = 0.040). The incidence of GU and GI toxicity did not correlate with the directions or size of the BAT moves. Regression analysis revealed that for acute GI toxicity, the only variable reaching statistical significance was BAT use; no variable, including BAT use, reached statistical significance for acute GU toxicity. CONCLUSIONS: The use of the BAT system did not change the rate of acute GU toxicity but did reduce the rate of acute GI toxicity. PMID- 15780366 TI - Evaluation of analgesic requirements and postoperative recovery after radical retropubic prostatectomy using long-acting spinal anesthesia. AB - OBJECTIVES: To analyze the postoperative pain, analgesic requirements, and convalescence of patients undergoing radical retropubic prostatectomy (RRP) under spinal anesthesia using long-acting morphine sulfate as preemptive analgesia. METHODS: A total of 103 consecutive men underwent RRP by a single surgeon. The time to tolerate oral fluids, time to unassisted ambulation, postoperative pain levels (visual analog pain score of 0 to 10), and analgesic requirements expressed in morphine equivalents were evaluated. Baseline patient characteristics and intraoperative factors (operating room time, blood loss) were also evaluated. RESULTS: The mean time to tolerate oral fluids and unassisted ambulation was 11.3 +/- 7.6 hours and 20 +/- 6 hours, respectively. The mean narcotic requirements were 7.4 +/- 6.1 morphine equivalents before discharge and 28.5 +/- 25.9 morphine equivalents in the first week after discharge. The mean visual analog pain score was 4.5 +/- 2.1 at discharge and fell significantly to 1.5 +/- 1.0 by the time of Foley catheter removal on postoperative day 7 or 8. The analgesic requirements after discharge correlated with the pain score at discharge (P = 0.016). The mean time to resumption of normal preoperative activities was 19.4 +/- 9.4 days. Two patients developed postspinal anesthesia headache. No other complications attributable to the anesthetic occurred. CONCLUSIONS: RRP may be performed through a small modified Pfannenstiel incision under spinal anesthesia containing long-acting morphine with little postoperative pain, low narcotic requirements, and a short convalescence. A prospective, randomized study is needed to compare the early postoperative outcomes of RRP performed using general versus spinal anesthesia. PMID- 15780367 TI - Does anesthesia method affect implant-induced prostate swelling? AB - OBJECTIVES: To investigate the impact of anesthesia selection on prostate gland swelling, acute toxicity, and implant quality. The outcome of prostate brachytherapy is dependent on the dose intensity and distribution. Preoperative and intraoperative planning are intended to optimize radiation delivery, but do not account for the impact of postoperative swelling on interseed spacing. Factors that increase swelling can be expected to increase the disparity between the intended and actual dose delivery. General anesthesia has been implicated in increased intraoperative bleeding during prostate surgery. METHODS: All iodine prostate implants planned and performed by the same radiation oncologist during a defined period were retrospectively reviewed. Excluded from the study were patients who had undergone preimplantation external beam radiotherapy or androgen deprivation. The remaining cases were analyzed to determine any association between the anesthesia type (general or spinal) and an increase in gland volume (from mapping transrectal ultrasonography to immediate postoperative computed tomography), implant quality (dosimetrically determined by minimal dose received by 90% of the volume [D90] and volume receiving 100% of prescribed dose [V100]), and acute toxicity (urinary retention, perineal/scrotal bruising). RESULTS: A total of 83 implants met the inclusion criteria. The outcomes did not significantly differ in regard to the median volume increase (23% versus 23.5%), D90 (115% versus 113%), V100 (97% versus 96.5%), acute urinary retention (3% versus 4%), or incidence or severity of perineal or scrotal bruising. No correlation was found between anesthesia type and any of the studied outcomes. CONCLUSIONS: Although only a prospective, randomized trial can definitively answer the question, our results suggest that the anesthesia selection for prostate brachytherapy does not influence prostate swelling, acute toxicity, or implant dosimetric quality. PMID- 15780368 TI - Recovery of quality of life in year after laparoscopic or retropubic radical prostatectomy: a multi-institutional longitudinal study. AB - OBJECTIVES: To compare the general and disease-specific health-related quality of life (HRQOL) after laparoscopic radical prostatectomy (LRP) with that after retropubic radical prostatectomy (RRP). METHODS: A total of 45 patients who underwent LRP alone and 121 who underwent RRP alone were prospectively enrolled in an HRQOL survey. We measured the general and disease-specific HRQOL with the Medical Outcomes Study 36-Item Short Form and University of California, Los Angeles, Prostate Cancer Index, respectively. The participants were asked to complete the questionnaires before and 1, 3, 6, and 12 months after surgery. RESULTS: No significant differences were found in the preoperative characteristics of the two groups. Repeated measures of analyses of variance revealed significantly different patterns of alteration in the several general HRQOL domains between the RRP and LRP groups. The LRP group tended to have a more delayed recovery than the RRP group in the domain of urinary function and bother. The sexual function and bother of both groups showed a substantially lower score throughout the postoperative period. When the LRP group was divided into two groups according to the surgical period, an apparent improvement in HRQOL was observed in the most recent LRP series. CONCLUSIONS: The two approaches showed different patterns of alteration regarding general HRQOL for 1 year after surgery. The LRP group reported delayed recovery of urinary and sexual function, which seemed to affect their general HRQOL. LRP appears to be still an evolving procedure. PMID- 15780369 TI - Intussusception of bladder neck improves early continence after radical prostatectomy: results of a prospective trial. AB - OBJECTIVES: To evaluate the impact of intussusception of the bladder neck on post radical prostatectomy incontinence. METHODS: A total of 272 men with organ confined prostate cancer who had undergone radical retropubic prostatectomy were studied. Of the 272 men, 139 underwent bladder neck intussusception and 133 did not. Patients completed validated questionnaires IIQ-7 and a symptom inventory. Continence was defined as the use of no or only one pad daily. RESULTS: Of the 272 men, 100%, 98.5%, and 96% answered the questionnaire and urinary symptom inventory at baseline and 3 and 12 months postoperatively, respectively. According to the protective pad requirement, 100%, 60%, and 86% of patients without intussusception and 100%, 77%, and 83% of the patients with intussusception were continent at baseline and 3 and 12 months postoperatively, respectively. Univariate logistic regression analysis revealed a statistically significant impact of intussusception on postprostatectomy incontinence at 3 months (P = 0.009), although continence at 12 months did not differ significantly (P = 0.5). CONCLUSIONS: Intussusception of the bladder neck had a significant effect on regaining continence at 3 months, although continence at 12 months was not affected. PMID- 15780370 TI - Predictors of short postoperative prostate-specific antigen doubling time for patients diagnosed during PSA era. AB - OBJECTIVES: To determine the preoperative and postoperative predictors of a short prostate-specific antigen (PSA) doubling time (PSADT) after radical prostatectomy for patients diagnosed during the PSA era. METHODS: Between 1989 and 2003, 1785 men underwent radical prostatectomy for 2002 American Joint Committee on Cancer (AJCC) Stage T1c or T2 prostate cancer. Of these men, 205 had documented PSA failure. The PSADT was calculated by assuming first-order kinetics and using a minimum of two detectable postoperative PSA measurements after a previous undetectable level. Multivariable logistic regression analyses were performed to determine the significant preoperative and postoperative predictors of a PSADT of less than 6 months. RESULTS: Patients with a greater biopsy Gleason score (P = 0.006), greater preoperative risk group (P = 0.002), greater prostatectomy Gleason score (P = 0.0006), greater 2002 AJCC pathologic stage (P = 0.01), or shorter time to postoperative PSA failure (P = 0.04) were more likely to have a shorter PSADT. Using multivariable analysis, high-risk disease (P = 0.0001) was the only preoperative factor that remained an independent significant predictor of a PSADT of less than 6 months. Of the postoperative factors, a prostatectomy Gleason score of 8 to 10 (P = 0.002), 2002 AJCC pathologic Stage T3b (P = 0.03), and time to PSA failure of less than 2 years (P = 0.05) remained significant independent predictors of a PSADT of less than 6 months. CONCLUSIONS: High-risk disease preoperatively and a prostatectomy Gleason score of 8 to 10, seminal vesicle invasion, or a time to PSA failure of less than 2 years postoperatively were significant independent indicators of developing a postoperative PSADT of less than 6 months. For these men, trials studying systemic therapy in addition to radical prostatectomy are needed. PMID- 15780371 TI - Urinary/serum prostate-specific antigen ratio: comparison with free/total serum prostate-specific antigen ratio in improving prostate cancer detection. AB - OBJECTIVES: To test the previously reported hypothesis, that the urinary/total serum (U/S) prostate-specific antigen (PSA) ratio improves the detection of prostate cancer, by evaluating the clinical usefulness of the U/S PSA ratio and comparing it with the free/total (F/T) serum PSA ratio. METHODS: A total of 165 patients undergoing transrectal ultrasound-guided prostate biopsy were prospectively included in this multicenter study. In all patients, PSA was measured from preoperative serum and 12-hour urine specimens in a centralized laboratory. RESULTS: Prostate cancer was identified in 83 of 165 patients. The differences between patients with and without prostate cancer were statistically significant (P <0.001) when considering the total PSA value (median, 10.2 ng/mL and 6.6 ng/mL respectively), F/T serum PSA ratio (0.11 and 0.18), and U/S PSA ratio (1.2 and 4.2). In the group of 79 patients with a PSA level between 4 and 10 ng/mL, receiver operating characteristic curves showed that the U/S PSA ratio was associated with a larger area under the curve (0.63; 95% confidence interval, 0.51 to 0.73) than the total PSA value (0.55; 95% confidence interval, 0.43 to 0.66) or F/T serum PSA ratio (0.60; 95% confidence interval, 0.49 to 0.71). The U/S PSA ratio did not correlate with patient age or prostate volume. CONCLUSIONS: Our results confirmed that the U/S PSA ratio may be a useful test in prostate cancer detection when the total serum PSA level is between 4 and 10 ng/mL. The F/T serum PSA ratio and U/S PSA ratio did not correlate. This suggests that these two tests could complement each other. PMID- 15780372 TI - High-grade prostatic intraepithelial neoplasia in needle biopsy as risk factor for detection of adenocarcinoma: current level of risk in screening population. AB - OBJECTIVES: To assess the current incidence of prostate carcinoma detection in serial biopsies in a prostate-specific antigen-based screening population after a diagnosis of isolated high-grade prostatic intraepithelial neoplasia (HG-PIN) in needle biopsy tissue. METHODS: We retrospectively identified 190 men with a diagnosis of isolated HG-PIN in needle biopsy tissue. Most men (86%) were diagnosed from 1996 to 2000. Logistic regression analysis was used to predict the presence of carcinoma in these 190 men and in a control group of 1677 men with only benign prostatic tissue in needle biopsy tissue. RESULTS: The cumulative risk of detection of carcinoma on serial sextant follow-up biopsies was 30.5% for those with isolated HG-PIN compared with 26.2% for the control group (P = 0.2). Patient age (P = 0.03) and serum prostate-specific antigen level (P = 0.02) were significantly linked to the risk of cancer detection, but suspicious digital rectal examination findings (P = 0.1), the presence of HG-PIN (P = 0.2), and the histologic attributes of PIN were not (all with nonsignificant P values). HG-PIN found on the first repeat biopsy was associated with a 41% risk of subsequent detection of carcinoma compared with an 18% risk if benign prostatic tissue was found on the first repeat biopsy (P = 0.01). CONCLUSIONS: The results of our study have shown that the current level of risk for the detection of prostate carcinoma in a screened population is 30.5% after a diagnosis of isolated HG-PIN in a needle biopsy. This risk level is lower than the previously reported risk of 33% to 50%. HG-PIN is a risk factor for carcinoma detection only when found on consecutive sextant biopsies. The data presented here should prompt reconsideration of repeat biopsy strategies for HG-PIN, and re-evaluation of the absolute necessity of repeat biopsy for all patients with HG-PIN. PMID- 15780374 TI - Evidence suggesting PSA cutpoint of 2.5 ng/mL for prompting prostate biopsy: review of 36,316 biopsies. AB - OBJECTIVES: To determine whether a prostate-specific antigen (PSA) level of 2.0, 2.5, or 4.0 ng/mL is the most appropriate cutpoint for determining the need for prostate biopsy. It has been suggested that the PSA cutpoint of 4.0 ng/mL is inappropriate because the rate of prostate cancer detection is similar in patients with lower PSA values. Some investigators have recommended a 2.6-ng/mL cutpoint. Others have recommended a cutpoint of 2.0 ng/mL. METHODS: A total of 36,316 prostate biopsies submitted to DIANON Systems from January 1, 1997 through December 31, 2001 were reviewed. These biopsy specimens also had DIANON PSA test results available that had been performed within 6 months of the biopsy date. These biopsies were stratified according to the PSA level within the 6 months before the time of biopsy, and the prostate cancer detection rate was determined for the stratified PSA levels. RESULTS: The detection rate of prostate cancer varied according to the PSA level. The incidence of prostate cancer was similar for the groups with less than 2.0 ng/mL and 2.0 to 2.5 ng/mL (18.67% and 21.89%, respectively). Also, the groups with 2.5 to 4.0 ng/mL and 4.0 to 10.0 ng/mL had similar cancer detection rates (27.48% and 30.08%, respectively). CONCLUSIONS: The prostate cancer detection rate for a PSA level between 2.5 and 4.0 ng/mL was similar (27.48%) to that for the PSA range of 4.0 to 10.0 ng/mL (30.08%). The absolute cutpoint used to determine the need to evaluate a patient for prostate cancer by biopsy is not clear; however, many studies have suggested that 2.5 ng/mL may be a more appropriate cutpoint than 4.0 ng/mL. PMID- 15780373 TI - Docetaxel-based chemotherapy as second-line treatment for paclitaxel-based chemotherapy-resistant hormone-refractory prostate cancer: a pilot study. AB - OBJECTIVES: To examine docetaxel (DTX)-based chemotherapy in paclitaxel (PTX) based chemotherapy-resistant hormone-refractory prostate cancer (HRPC) to investigate the activity and degree of toxicity in a pilot study. We have previously reported on the effectiveness of combination chemotherapy with PTX, estramustine (EMP), and carboplatin against HRPC. Although many patients with HRPC initially responded to this PTX/EMP-based combination chemotherapy, most finally progressed to PTX-resistant status within a mean of less than 1 year. DTX is an inhibitor of microtubule depolymerization like PTX and has demonstrated activity against PTX-resistant metastatic tumors. METHODS: The subjects were 15 patients with HRPC who displayed disease progression while receiving PTX/EMP/carboplatin combination chemotherapy. The patients were treated with intravenous DTX 30 mg/m2 weekly, oral EMP 10 mg/kg daily, and intravenous carboplatin (dosed to an area under the curve of 6) on day 1 of every 4-week cycle. Treatment was continued until disease progression or excessive toxicity. RESULTS: Each patient received a median of eight consecutive cycles. The prostate specific antigen level decreased by 50% or more in 33.3% of patients and by 90% or more in 13.3%. The median follow-up was 49.6 weeks, with median time to progression of 25.0 weeks and median overall survival of 54.0 weeks. One patient died of interstitial pneumonitis. One patient developed secondary osteomyelodysplastic syndrome. The major severe toxicities were grade 3 or 4 anemia in 66.7% of patients, leukopenia in 26.7%, and thrombocytopenia in 40.0%. CONCLUSIONS: The results of our study have shown that DTX is comparatively active for heavily pretreated patients with PTX-resistant HRPC. However, given the significant toxicities and small subject population, well-designed Phase I-II trials of the improved regimen are warranted. PMID- 15780375 TI - Benefits of microsurgical repair of adolescent varicocele: comparison of semen parameters in fertile and infertile adults with varicocele. AB - OBJECTIVES: To compare preoperative semen parameters and the effect of microscopic varicocele repair on semen parameters in adolescents, fertile adults, and infertile adults with varicocele. METHODS: A total of 96 patients (35 adolescents, 20 fertile adults, and 41 infertile adults) were included in the primary analysis. RESULTS: The fertile adults had a higher sperm count than did the adolescents or infertile adults (P <0.001). In infertile adults, the percentage of normal sperm increased from 14.9% +/- 1.0% preoperatively to 21.2% +/- 2.2% at 3 months (P = 0.013) and 22.1% +/- 2.2% (P = 0.003) at 6 months. The infertile adults had a lower sperm count than did the adolescents and fertile adults at both 3 (P = 0.014) and 6 (P = 0.003) months. The postoperative sperm motility in adolescents was significantly greater than that in the infertile adults at 3 (P = 0.009) and 6 (P = 0.003) months. The proportion of sperm with normal morphology was not statistically different at either 3 or 6 months among the groups. No significant differences were detected in the increase in semen parameters at 3 or 6 months among patients before or after surgery. CONCLUSIONS: The results of our findings suggest that after varicocele repair the degree of improvement in semen parameters is not different. However, because the baseline values of the semen parameters in adolescents were better than those in infertile adults, the postoperative values in the adolescents were superior to those in infertile adults and at least as good as those in fertile adults. PMID- 15780376 TI - High prevalence of erectile dysfunction in young male patients after intramedullary femoral nailing. AB - OBJECTIVES: To evaluate, given the central role of the pudendal nerves in erection, the impact of potential infraclinical lesions on male sexual function. After intramedullary femoral fixation, countertraction on the fracture table has sporadically been involved in pudendal neurapraxia. Patients with tibial fractures served as controls. METHODS: A total of 168 patients treated for femoral or tibial shaft fractures by intramedullary nailing were mailed the International Index of Erectile Function questionnaire, which addresses all aspects of male sexual function and permits grading of the severity of erectile dysfunction (ED). Univariate and multivariate analyses were conducted to test for factors associated with ED. RESULTS: Of the 168 patients, 101 (60.1%) returned the questionnaire. A greater proportion of ED was observed in sexually active patients after femoral fracture than after tibial fracture (40.5% versus 12.5%, P <0.01). The differential prevalence of ED in both groups subjected to comparable high-energy trauma suggested that post-traumatic stress disorder was of marginal importance in ED occurring after femoral nailing. Greater intraoperative doses of curare were associated with better sexual functioning in sexually active patients after femoral fracture (10.6 versus 7.5 mg in patients without and with ED, respectively, P = 0.02), suggesting that postoperative ED could be partially prevented by optimal muscle relaxation during fracture reduction. CONCLUSIONS: Erectile dysfunction was shown to be highly prevalent after intramedullary nailing of femoral shaft fractures. Greater intraoperative curare doses, resulting in optimal relaxation and reduced pressure on the pudendal nerves by the perineal post, were associated with better sexual functioning. PMID- 15780377 TI - Subureteral injection of Deflux for correction of reflux: analysis of factors predicting success. AB - OBJECTIVES: To review, prospectively, our experience with endoscopic Deflux injection and evaluate the volume injected, grade, endoscopic appearance after injection, and presence or absence of voiding dysfunction as predictors of success. Subureteral injection of dextranomer/hyaluronic acid copolymer (Deflux) has become an effective treatment of vesicoureteral reflux. METHODS: A total of 52 patients (50 females and 2 males; 80 ureters) were treated with a single subureteral injection of Deflux. The mean patient age was 7.6 years (range 14 months to 22 years). The presence or absence of voiding dysfunction was evaluated with a preoperative questionnaire and patient history. The volume of Deflux injected in each ureter was recorded. The endoscopic appearance after injection was recorded as "volcano" or "other." Success was defined as no reflux on postoperative voiding cystourethrography. RESULTS: The success rate by grade of reflux in individual ureters was 82%, 84%, 78%, and 73% for grade 1, 2, 3, and 4 vesicoureteral reflux, respectively. No statistically significant difference was found in the cure rate by grade (P = 0.76). The overall cure rate by ureter was 80% and by patient was 71%. New contralateral reflux developed in 12.5% of patients. No statistically significant difference was found in the cure rate with respect to the volume injected or the presence or absence of voiding dysfunction. The ureteral cure rate with volcano and alternate morphology was 87% and 53%, respectively (P = 0.004). CONCLUSIONS: Mound morphology was the only statistically significant predictor of a successful outcome, with an associated cure rate of 87%. Concomitant voiding dysfunction did not have an adverse effect on the cure rate. In our experience, no statistically significant difference was found in the cure rate for grades 1 through 4 vesicoureteral reflux after a single injection of Deflux. PMID- 15780378 TI - Intractable high-pressure bladder in female infants with spina bifida: clinical characteristics and use of vesicostomy. AB - OBJECTIVES: To characterize the clinical characteristics of a group of children with neurogenic bladder, high leak point pressures (LPPs), bladder wall changes, and hydronephrosis in whom medical management failed, who then underwent vesicostomy. Medical management can, rarely, fail in children with neurogenic bladder, high LPPs, bladder wall changes, and hydronephrosis. When it does, surgical intervention is necessary. METHODS: Between 1988 and 2001, 8 girls with spina bifida (4 with thoracic and 4 with lumbosacral) underwent vesicostomy after medical management. All had undergone neonatal back closure and five had ventriculoperitoneal shunts. The mean follow-up after vesicostomy was 6 years. RESULTS: High-grade reflux was present at birth in 3 patients and developed later in 3 patients at 1 year. Two had undergone ureteral reimplantation before vesicostomy. Four girls had trabeculation at birth and it developed in three at 1 year and one at 3 years. Hydronephrosis was present at birth in 2 patients and developed later in 6. High LPPs were noted at younger than 10 months of age in 6 patients. Oral oxybutynin was begun in 7 patients before 1 year of age and 5 were later given intravesical oxybutynin. Six patients began clean intermittent catheterization in infancy. The indications for vesicostomy were persistently elevated LPPs and hydronephrosis. Six had recurrent urinary tract infections. The mean age at vesicostomy was 3 years. Hydronephrosis resolved in 7 patients, and only 1 had additional febrile urinary tract infections. Two underwent reversal along with bladder augmentation and were continent at last follow-up. CONCLUSIONS: A small subset of girls with spina bifida are unresponsive to early and intensive medical therapy. Most have high LPPs and bladder wall and upper tract changes early in infancy. Vesicostomy reverses the clinical course. At an appropriate age, these children can undergo reconstruction with a high likelihood of continence. PMID- 15780379 TI - Simultaneous bilateral retrograde intrarenal surgery for bilateral complex upper tract stone disease. AB - Recent advances in retrograde endoscopy have greatly expanded the role of minimally invasive surgery in addressing upper tract stone disease. In an attempt to decrease patient morbidity further, we present our initial experience with simultaneous bilateral retrograde intrarenal surgery in a patient with complex bilateral upper tract stones. PMID- 15780380 TI - Laparoscopic retroperitoneal lymph node dissection: duplication of open technique. AB - Laparoscopic retroperitoneal lymph node dissection has been used to stage germ cell testicular cancer. Since its initial description, this minimally invasive procedure has evolved into a therapeutic operation that adheres to established strict oncologic principles. A modified template dissection that fully duplicates the open technique is now routinely performed at our institution. We describe and show in the accompanying video segments a laparoscopic modified template dissection. PMID- 15780381 TI - Open partial nephrectomy using saline-enhanced monopolar radiofrequency device: evaluation of novel surgical technique with TissueLink DS3.0 Dissecting Sealer. AB - INTRODUCTION: Nephron-sparing surgery has emerged as the preferred surgical management of select renal masses. Advancements in surgical and hemostatic techniques, together with long-term data on cancer-free survival, have led to greater use of these procedures. The TissueLink Dissecting Sealer is a new device that couples radiofrequency energy with low-volume saline irrigation to give improved hemostasis during dissection. We assessed this device as an alternative dissecting tool during open partial nephrectomy. TECHNICAL CONSIDERATIONS: A total of 32 consecutive patients underwent open partial nephrectomy during a 20 month period. These masses were resected using the TissueLink Dissecting Sealer or the surgeon's conventional method of dissection (needlepoint electrocautery). Perioperative parameters, including operative time, tumor size, blood loss, and hilar vessel occlusion times, were retrospectively reviewed and compared. The average age in the TissueLink group (n = 16) was 60 years (range 46 to 87) compared with 61 years (range 45 to 89) in the conventional group (n = 16). The average tumor size in the TissueLink group was 3.1 +/- 1.4 cm (range 1.2 to 7.0) compared with 2.5 +/- 1.3 cm (range 0.8 to 6.0) in the conventional group. The hilar vessels were clamped in select patients (with similar occlusion times) from both groups because of deep tumor penetration or a central location. The average blood loss was 126 +/- 40 mL (range 50 to 200) for the TissueLink group versus 194 +/- 105 mL (range 50 to 400) in the conventional group, a statistically significant difference (P = 0.027). No perioperative complications occurred. CONCLUSIONS: TissueLink-assisted open partial nephrectomy is a novel technique that couples radiofrequency energy with low-volume saline irrigation to provide excellent hemostasis during and after renal tumor dissection. PMID- 15780382 TI - Technique for removal of symptomatic bone anchors placed during stress incontinence surgery. AB - INTRODUCTION: To present an uncomplicated, reliable technique for bone anchor removal in patients with anchor-related infections or chronic pain. TECHNICAL CONSIDERATIONS: We removed 17 anchors from 9 patients between 1999 and 2004. The surgical technique used fluoroscopy for localization of the anchors and an orthopedic broken screw removal instrument for resection of the anchor and surrounding bone. Nine patients with bone anchors had been symptomatic with chronic pain and/or wound drainage for a mean of 23.7 months before surgery. After removal, 10 anchors grew positive bacterial cultures. The most common organism was coagulase-negative Staphylococcus. During a mean follow-up period of 6.8 months after hardware removal, 8 of the 9 patients had symptomatic improvement. CONCLUSIONS: Fluoroscopic localization and en bloc resection with a broken screw removal instrument is an effective method of removing symptomatic bone anchors. PMID- 15780383 TI - Urinary retention and obstructive nephropathy secondary to mullerian duct cyst in an adult. PMID- 15780384 TI - Lymphangioleiomyomatosis presenting as bladder outlet obstruction. PMID- 15780385 TI - Abdominoperineal excision of male lower urinary tract for synchronous adenocarcinoma of urethra and urinary bladder. AB - Urethral adenocarcinoma is the least common histologic subtype of a rare primary carcinoma of the male urethra. Its site of origin remains speculative. The scarcity of reported cases in the literature makes it difficult to define the optimal management. This report is of a 40-year-old morbidly obese man with synchronous adenocarcinoma of the bulbar urethra and bladder, treated surgically by en bloc abdominoperineal penoprostatocystectomy. The pleural recurrence, after a disease-free period of 2 years, responded well to platinum-docetaxel-based systemic chemotherapy. Synchronous occurrence of adenocarcinoma of the urethra and bladder has not been previously reported. This case also highlights the possible role of upcoming adjuvant chemotherapeutic agents. PMID- 15780386 TI - Endoscopic seminal vesicle stone removal. AB - Seminal vesicle stones are extremely rare, and few cases have been reported. Treatment requires removal of the stone, generally through an open vesiculectomy. A 31-year-old man presented with perineal pain, painful ejaculation, and infertility of several years' duration. Multiple stones in the seminal vesicle duct system were diagnosed by radiologic examination. We treated the patient by seminal vesicle endoscopic stone removal, thereby obviating organ loss. The composition of the stones was whewellite. To our knowledge, this approach has not been previously reported, and our result may be encouraging for treatment of such pathologic conditions of the seminal vesicles. PMID- 15780387 TI - Testicular metastasis 10 years after resection of appendiceal carcinoid. AB - Testicular carcinoids are very rare and account for less than 1% of all testicular neoplasms. They may present as primary testicular tumors or secondary tumors from extratesticular sources. We report a case of a secondary testicular carcinoid occurring 10 years after surgical treatment of an appendiceal carcinoid. The patient underwent radical orchiectomy and at 24 months of follow up had no evidence of metastases elsewhere. Because secondary carcinoids have a worse clinical course and prognosis than primary tumors, metastatic disease should be excluded before the tumor is identified as a primary. Long-term follow up is necessary for patients with carcinoid tumor owing to its indolent course and the risk of metastasis several years after treatment of the primary. PMID- 15780388 TI - Epididymo-orchitis and testicular abscess due to Nocardia asteroides complex. AB - Nocardia asteroides complex is a rare human pathogen chiefly affecting immunosuppressed patients. We present the sixth case of epididymo-orchitis attributable to this organism. A 78-year-old man presented to the Mayo Clinic with fever and weight loss after being treated with cyclophosphamide and prednisone for an immune-mediated vasculitis. He was found to have a testicular abscess, and orchiectomy was performed. N. asteroides complex was identified on the wound cultures. Sulfa therapy was begun. In previous case reports, this condition carried a 50% mortality rate. Patients who survive seemed to benefit from aggressive surgical debridement and long-term sulfa therapy. PMID- 15780389 TI - The kidney bridge procedure for bilateral ureteral loss. AB - Bilateral ureteral loss is an uncommon occurrence that challenges the usual reconstructive approaches. We developed a novel procedure, using autotransplantation of one kidney to bridge the ureteral deficit of the contralateral kidney. A 61-year-old woman developed long bilateral mid and distal ureteral strictures from idiopathic retroperitoneal fibrosis. The left kidney was autotransplanted to the right iliac fossa and the right ureter was anastomosed to the autotransplanted kidney's pelvis, followed by ureteroneocystostomy, all over a stent. This provided an intestine-free, physiologically normal system for draining both kidneys into the bladder. She had a stable creatinine of 1.7 mg/dL at 18 months postoperatively. PMID- 15780390 TI - Plasma renin activity revealed renal artery stenosis concealed by aneurysms. AB - We describe a case of renovascular hypertension with renal artery stenosis concealed by aneurysms. Arteriography demonstrated no apparent renal artery stenosis, but did reveal aneurysms on the left renal artery. Captopril-loaded renoscintigraphy could not detect disturbed renal perfusion. High basal and exaggerated plasma renin activity after captopril administration were the only clues indicating renovascular hypertension. A reduction of the systemic blood pressure and normalized plasma renin activity after resection of the aneurysms confirmed preoperative renovascular hypertension. Fibromuscular dysplasia was an underlying cause of the arterial deformity. In cases of hypertension accompanied by renal artery aneurysms, the captopril-challenge test can be a useful tool to detect renal artery stenosis concealed by the aneurysms. PMID- 15780391 TI - Page kidney as a complication of percutaneous antegrade endopyelotomy. AB - A kidney with a compressive process that results in hyperreninemic hypertension is often referred to as a "Page" kidney. We present a case of "Page" kidney that resulted as a complication of percutaneous antegrade endopyelotomy. The patient had a typical presentation and classic radiologic signs of such a process. Surgical management consisted of nephrectomy, with resultant resolution of signs and symptoms. PMID- 15780392 TI - Neuromuscular complications after percutaneous radiofrequency ablation of renal tumors. AB - Neuromuscular complications after percutaneous renal tumor ablation have not been previously reported. In a series of 48 patients undergoing percutaneous ablation, 3 of our patients had neuromuscular complications. One patient developed permanent flank laxity and two had transient paresthesias. Neuromuscular complications are uncommon, but may occur with percutaneous renal ablative surgery. PMID- 15780393 TI - Methicillin-resistant Staphylococcus aureus endocarditis after transurethral prostatic resection. AB - We report the first case, to our knowledge, of methicillin-resistant Staphylococcus aureus endocarditis complicating transurethral prostatic resection for benign prostatic hyperplasia. The patient had been catheterized preoperatively because of urinary retention and, postoperatively, developed pyrexia with persistent methicillin-resistant S. aureus-positive blood cultures. The cause remained elusive, despite extensive investigations (including echocardiography), until he developed embolic phenomena pathognomonic of infective endocarditis 1 month into treatment with intravenous vancomycin. PMID- 15780394 TI - Treatment of postpriapism erectile dysfunction by closure of persistent distal glans-cavernosum fistulas 5 years after shunt creation. AB - Erectile dysfunction is common after the formation of a glans-cavernosum shunt for intractable priapism. Often patients require placement of a penile prosthesis to restore sexual function. We report a case of restoration of full erections after reversal of a glans-cavernosum shunt in a 30-year-old man who had been treated for priapism 5 years previously. On the basis of our experience, it seems reasonable to attempt shunt reversal for correction of erectile dysfunction in selected patients who likely have a persistent vascular fistula. PMID- 15780395 TI - Bilateral testicular tumors in an infertile man previously treated with follicle stimulating hormones. AB - A 43-year-old infertile man with oligozoospermia and normal serum gonadotropin and cytogenetic findings was treated with follicle-stimulating hormone for 1 year at our institution. Two years later, the patient presented with bilateral palpable testicular tumors. His beta-human chorionic gonadotropin and lactate dehydrogenase levels were elevated. Pathologic examination disclosed a pure seminoma in each testis (pT1N0M0). Adjuvant radiotherapy was administered. The tumors might have been induced by follicle-stimulating hormone treatment. Careful follow-up examination of the testis is necessary in men who have received hormonal treatment for infertility. PMID- 15780396 TI - Fibroepithelial polyp of glans penis. AB - A case of fibroepithelial polyp of the penis in a 59-year-old man is presented. The physical examination showed that the lesion affected the ventral surface of the glans, near the urethral meatus. No communication was found with the urethra. The patient had a history of long-term condom catheter use. He was successfully treated by wide local excision. The histopathologic diagnosis was a fibroepithelial polyp. Postoperatively, the follow-up examinations at 3, 6, 12, and 24 months showed normal findings and no recurrence. PMID- 15780397 TI - Acute urinary retention in ileum conduit urinary diversion. AB - Three years after ileum conduit urinary diversion, a 68-year-old patient was referred to us with acute urinary retention. Ultrasound examination demonstrated an extremely dilated, urine-filled ileum conduit and dilated kidneys. Placing a catheter into the subfascial part of the conduit to drain the urine was impossible because of extreme kinking and compression of the conduit in the fascial level. Ultrasound-guided puncture of the conduit was performed, and 170 mL of urine was aspirated. Afterward, a 16F catheter was easily placed. Clear urine drained. At last follow-up, no further episodes of urinary retention had occurred. PMID- 15780398 TI - Acute obstructive renal failure after laparoscopic cryoablation of multiple renal tumors in a solitary kidney. AB - A 67-year-old man with a solitary left kidney and baseline serum creatinine of 3.3 ng/mL (normal range 0.7 to 1.4) underwent laparoscopic cryoablation of two enhancing, solid renal tumors. Postoperatively, the case was complicated by a large blood clot in the renal collecting system causing obstruction and anuria. This complication of cryotherapy, never before reported to our knowledge, was treated successfully with a temporary ureteral stent. Follow-up magnetic resonance imaging at 3 months did not demonstrate any new lesions or enhancement of the treated renal masses. His serum creatinine at 3 months was 3.7 ng/mL. PMID- 15780399 TI - Ligands for peroxisome proliferator-activated receptor gamma have potent antitumor effect against human renal cell carcinoma. AB - OBJECTIVES: To examine whether peroxisome proliferator-activated receptor gamma (PPARgamma) is expressed in human renal cell carcinoma (RCC) cells, and whether activation of PPARgamma by its ligands can have multiple antitumor effects on human RCC cells in vitro. METHODS: We examined the expression of PPARgamma in four human RCC cell lines by reverse transcriptase-polymerase chain reaction and immunocytochemical staining. The effects of two PPARgamma ligands, pioglitazone and 15-deoxy-Delta12,14-prostaglandin J2, on cell proliferation were investigated by 3-[4,5-dimethylthiazol-2-thiazoly]-2,5-diphenyltetrazolium bromide assay. The induction of apoptosis by the ligands was examined using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method and Annexin V assay. Furthermore, we investigated whether these ligands suppressed the production of angiogenic factors, vascular endothelial growth factor and basic fibroblast growth factor, by enzyme-linked immunosorbent assay. RESULTS: PPARgamma and retinoid X receptor, which forms a heterodimer with PPARgamma, were expressed in all RCC cell lines. In addition, immunocytochemical studies showed expression of PPARgamma protein in the RCC cells. PPARgamma ligands inhibited the cell growth in all cells in a dose-dependent manner. These ligands also induced apoptosis. Furthermore, secretion of both vascular endothelial growth factor and basic fibroblast growth factor was inhibited by these ligands in a dose-dependent and time-dependent manner. CONCLUSIONS: Ligands for PPARgamma have multiple antitumor effects in human RCC cells in vitro. Activation of the PPARgamma pathway may be a new strategy for treatment of patients with RCC. PMID- 15780400 TI - Canine model of surgical stress response comparing standard laparoscopic, microlaparoscopic, and hand-assisted laparoscopic nephrectomy. AB - OBJECTIVES: To compare the postoperative stress induced by standard laparoscopic, microlaparoscopic, and hand-assisted laparoscopic (HALS) nephrectomy in an animal model. METHODS: A total of 39 dogs underwent standard laparoscopic (n = 19), microlaparoscopic (n = 11), or HALS (n = 9) left nephrectomy. The serum cortisol levels were measured preoperatively, at skin closure, and 2, 4, and 6 hours postoperatively. RESULTS: Compared with the preoperative level, a sharp rise was noted in the serum cortisol taken at skin closure that correlated with both operative time (P = 0.003) and method (P = 0.009 for HALS versus microlaparoscopy and P = 0.02 for HALS versus standard). HALS had the lowest cortisol increase and shortest operative time, and microlaparoscopy had the greatest cortisol increase and longest operative time (P = 0.03 for cortisol increase, HALS versus microlaparoscopy). Two hours postoperatively, HALS was the only method associated with a continuing increase in cortisol levels (P = 0.01 and P = 0.02 compared with the other methods) and had the greatest cortisol level relative to baseline. The cortisol level decreased at all postoperative points in the other groups. By 4 hours, all methods were associated with similar and falling cortisol levels. CONCLUSIONS: HALS nephrectomy was associated with a greater operative stress response in the first 2 postoperative hours in dogs compared with standard laparoscopic and microlaparoscopic nephrectomy that was partially ameliorated by the lower intraoperative cortisol production in the more rapid HALS procedure. Microlaparoscopy provided no benefit in terms of reduced surgical stress. The stress differences among the techniques were insignificant by 4 hours postoperatively. PMID- 15780401 TI - Cyclic adenosine monophosphate and cyclic guanosine monophosphate phosphodiesterase isoenzymes in human vagina: relation to nitric oxide synthase isoforms and vasoactive intestinal polypeptide-containing nerves. AB - OBJECTIVES: To evaluate the distribution of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) phosphodiesterases (PDEs) in relation to nitric oxide synthase isoforms and vasoactive intestinal polypeptide (VIP) in specimens of the human vagina. Nitric oxide and VIP, mediating biologic signals through cGMP and cAMP, have been assumed to be involved in the control of vaginal smooth muscle. METHODS: Immunohistochemical techniques were applied to sectioned specimens of the human vaginal wall to evaluate the presence of the PDE isoenzymes 3, 4, 5, and 10 in relation to neuronal nitric oxide synthase (nNOS), endothelial NOS (eNOS), and VIP. RESULTS: Immunoreactivity (IR) for cAMP degrading PDE-4 was observed in the vaginal nonvascular smooth musculature, as well as in the wall of subepithelial arteries. VIP-IR nerves innervated the smooth muscle portion of the vaginal wall and also formed a subepithelial network. Immunoreactivity specific for PDE-5 was also registered in vascular and nonvascular vaginal smooth muscle. Immunosignals for eNOS were detected in the endothelial lining of arteries containing PDE-5-IR smooth muscle cells. These arteries were supplied by nNOS-IR nerve fibers. PDE-10-IR smooth muscle cells were located in muscle bundles of the vaginal wall. CONCLUSIONS: Our study revealed immunoreactivity specific for PDE-4, PDE-5, and PDE-10 in the vascular and nonvascular smooth muscle of the vagina. Immunosignals for PDE-4 and PDE-5 were also observed in close proximity to nNOS-IR or VIP-IR nerve fibers or to eNOS-IR endothelial cells. The distribution of PDEs may indicate a role of these enzymes in the control of the function of the human vagina. PMID- 15780402 TI - Free radical-scavenging activity of Korean red ginseng for erectile dysfunction in non-insulin-dependent diabetes mellitus rats. AB - OBJECTIVES: To investigate the antioxidant activity of Korean red ginseng (KRG) and its effect on erectile function in non-insulin-dependent diabetes mellitus (NIDDM) rats. Oxidative stress is an important factor in vascular complications of diabetes. METHODS: A total of 84 male Sprague-Dawley rats were included in this study. NIDDM was induced by an intraperitoneal injection of 90 mg/kg of streptozotocin on day 2 after birth. According to the diabetic period, they were classified as either short-term (22 weeks, n = 32) or long-term (38 weeks, n = 32) diabetics. Of those, 20 (10 short-term and 10 long-term) were fed 30 mg/kg of KRG three times weekly for 1 month. The remaining diabetic rats (22 short-term and 22 long-term) and their age-matched controls (n = 10 each for each group) were fed a normal diet. Erectile function was measured after electrostimulation of the cavernous nerve. The total cavernous malondialdehyde and glutathione levels were measured using a spectrophotometric assay. RESULTS: The intracavernous pressure after nerve stimulation and cavernous glutathione level were significantly lower in the long-term than the short-term diabetics with a normal diet and were markedly decreased compared with their age-matched controls (P <0.01 and P <0.05, respectively). The malondialdehyde content was markedly increased in the short-term diabetics compared with the controls (P <0.05). In contrast, erectile function was not impaired in the diabetic group treated with KRG. Furthermore, both glutathione and malondialdehyde levels in those treated with KRG were comparable to their age-matched controls. CONCLUSIONS: Oxidative stress to cavernous tissue may be a contributory factor in erectile dysfunction in diabetics. KRG may preserve potency in the NIDDM rats through its antioxidant activity. PMID- 15780403 TI - Preliminary study of immunomagnetic quantification of circulating tumor cells in patients with advanced disease. AB - OBJECTIVES: To enumerate the amount of circulating tumor cells (CTCs) in patients with advanced prostate cancer and to investigate the relationship between these numbers, prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSM) expression, and clinical parameters. METHODS: Whole blood was collected in proprietary CellSave tubes. Mononuclear cell fractions were isolated using epithelial cell antibody-coated magnetic nanoparticles. On one half of each immunomagnetically enriched cell fraction, automated fluorescent microscopy was used to identify the epithelial tumor cells. From the remainder of each sample, RNA extraction, cDNA synthesis, and polymerase chain reaction amplification of PSA and PSM were performed. RESULTS: Eighty-four patients with advanced prostate cancer submitted 130 samples for analysis. Intact CTCs were identified in 62% of samples; 83.3% of CTC-positive and 0% of CTC-negative samples were reverse transcriptase-polymerase chain reaction positive for PSA and PSM (P = 0.001). A significant positive correlation was found between the CTC number and PSA (r = 0.49), alkaline phosphatase (r = 0.47), and lactate dehydrogenase (r = 0.55) levels, and a significant negative correlation with hemoglobin (r = -0.35). The initial Gleason grade, prior therapy, current therapy, and type of metastasis (bone, soft tissue) did not correlate significantly with the CTC number. CONCLUSIONS: The presence of intact CTCs and the expression of PSA and PSM demonstrated robust agreement. The tumor cell numbers reflected current disease status and correlated significantly with the clinical disease indicators of PSA, hemoglobin, and liver function tests. These findings warrant further investigation of the diagnostic and prognostic value of enumerating intact CTCs. PMID- 15780404 TI - Effect of botulinum toxin on detrusor overactivity induced by intravesical adenosine triphosphate and capsaicin in a rat model. AB - OBJECTIVES: To assess the effects of intravesical injection of botulinum toxin type A (BTX) on a model of detrusor overactivity induced by intravesical infusions of adenosine triphosphate (ATP) and capsaicin. BTX has recently been used clinically to treat overactive bladder syndromes without a precise knowledge of the mechanism of action. METHODS: Twelve Sprague-Dawley rats underwent BTX injections. Six received 1.0 U and 6 received 0.5 U. BTX injections were done at bladder tube placement. Ten rats received saline injections as controls. After 48 hours of recovery, all 22 animals underwent awake, conscious cystometrography (CMG), performed using both saline and ATP (20 mM) intravesical infusion at 0.074 mL/min. In another 4 rats, capsaicin (100 microM) was infused intravesically before and after the BTX injections. The CMG parameters calculated included bladder contraction pressures and contraction frequencies (contractions per minute or Herz). RESULTS: Intravesical saline CMG produced a contraction frequency of 0.78 +/- 0.10 Hz. Intravesical ATP doubled this voiding frequency to 1.45 +/- 0.18 Hz (P = 0.003). BTX treatment at 1.0 U reduced the frequency to 0.91 +/- 0.13 Hz (P = 0.02). BTX injection significantly decreased the bladder contraction pressure during saline and ATP CMG. However, 0.5 U BTX did not decrease ATP-induced overactivity; therefore, in the capsaicin experiments, 1.0 U BTX was used. Although BTX tended to reverse detrusor overactivity secondary to intravesical capsaicin, this difference was not statistically significant. CONCLUSIONS: Intravesical infusion of either ATP or capsaicin can induce detrusor overactivity. BTX was more effective in blocking the effect of ATP than of capsaicin, although BTX injection did show a trend in reducing the contraction frequencies and amplitudes induced by capsaicin. The clinical utility of using BTX to treat overactive bladder syndromes and bladder hypersensory states, especially those that may be caused by an augmentation of the purinergic pathway, should be studied further. PMID- 15780405 TI - Plasma bubble formation induced by holmium laser. PMID- 15780406 TI - Applying complexed prostate-specific antigen to clinical practice. PMID- 15780410 TI - Nerve-sparing laparoscopic radical prostatectomy: replicating the open surgical technique. PMID- 15780411 TI - The interpretation of nucleic acid amplification tests for tuberculosis: do rapid tests change treatment decisions? AB - OBJECTIVES: To describe changes in treatment decisions after receipt of nucleic acid amplification (NAA) test for the diagnosis of M. tuberculosis. METHODS: Retrospective notes review of treatment decisions in patients receiving a NAA test for suspected pulmonary or non-pulmonary tuberculosis at the Royal Free Hospital in London between March 2001 and February 2002. Notes were sought on a 50% random sample of patients with both smear and NAA negative specimens and all patients with other specimen results. RESULTS: Two hundred and fifty patients were tested with NAA; clinical details were obtained on 138; 61 were ever treated. Seventeen (17/18) smear-negative patients were started on treatment after a positive NAA; none of six smear-negative patients treated prior to a negative NAA result had treatment stopped. Seventeen (17/21) smear-positive patients were treated prior to NAA result and all were NAA positive; treatment was delayed in four smear-positive patients until receipt of an NAA and one NAA negative patient was not treated. CONCLUSIONS: In routine practice a positive test in an untreated smear-negative patient leads to decision to treat in almost all, but the proportion testing positive is low (8% or 17/219). In patients already on treatment negative tests did not lead to decisions to stop. PMID- 15780412 TI - Tuberculous meningitis and miliary tuberculosis: the Rich focus revisited. AB - Tuberculous meningitis (TBM) develops most often when a caseating meningeal or sub-cortical focus, the Rich focus, discharges its contents into the subarachnoid space. It is recognized that TBM is frequently accompanied by miliary tuberculosis, but the relationship between the development of the Rich focus and miliary tuberculosis remains controversial. The original descriptions of Arnold Rich and Howard McCordock are reviewed together with the work of other pathologists and the observations of the natural history of tuberculosis by astute clinicians such as Arvid Wallgren and Edith Lincoln. Rich and McCordock dissociated miliary tuberculosis from a role in the pathogenesis of TBM, and this view continues to appear in reviews and textbooks dealing with TBM. We suggest, particularly in childhood, that miliary tuberculosis is indeed directly involved in the pathogenesis of TBM in as much as that the overwhelming bacillaemia that accompanies miliary tuberculosis serves to increase the likelihood that a meningeal or sub-cortical Rich focus will be established, which may in its turn caseate and give rise to TBM. PMID- 15780413 TI - Salvage therapy with caspofungin for invasive aspergillosis: results from the caspofungin compassionate use study. AB - OBJECTIVES: The objective was to prospectively assess the efficacy and safety of caspofungin as salvage therapy for invasive aspergillosis in patients enrolled in the caspofungin compassionate-use study. METHODS: Forty-eight patients with invasive Aspergillus infections (36 with pulmonary infection, 12 with extrapulmonary or disseminated infection) were enrolled in this study. All patients were refractory to or intolerant of intravenous amphotericin B or a lipid amphotericin formulation(s). Efficacy was assessed at end of intravenous caspofungin therapy based on the clinical (symptom/sign and radiographic) response. RESULTS: Underlying diseases included hematological malignancy (69%), organ transplant (8%), and AIDS (6%). Forty-three (90%) patients were refractory to prior antifungal treatment, including 25 patients refractory to multiple agents. Sixteen (33%) were neutropenic at study entry. Following caspofungin therapy, a favorable response was noted in 44% (20/45) of the patients, including nine (20%) and 11 (24%) patients with complete and partial responses, respectively. Caspofungin was generally well tolerated one serious drug-related adverse event was reported. CONCLUSIONS: In this study, caspofungin was an effective alternative for patients with refractory Aspergillus infections. PMID- 15780414 TI - Dalbavancin compared with vancomycin for prevention of Staphylococcus aureus colonization of devices in vivo. AB - OBJECTIVES: Although active against free-floating bacteria, vancomycin displays a poor activity against organisms embedded within the biofilm surrounding implanted devices. Dalbavancin is a novel glycopeptide antibiotic with strong activity against staphylococci and a long half-life that allows for once-a-week dosing. The objective of this animal study was to examine the ability of dalbavancin and vancomycin to prevent Staphylococcus aureus colonization of devices. METHODS: Twelve rabbits were randomized, in three groups of four each, to receive intravenous injections of dalbavancin, vancomycin or normal saline (control). Eight polyurethane catheter segments were subcutaneously implanted in the back of each rabbit, then inoculated with S. aureus. Rabbits were sacrificed a week later and explanted devices were cultured. RESULTS: The rates of device colonization were comparable in the vancomycin (53%) and control (47%) groups, whereas only 28% of devices in the dalbavancin group became colonized. There was a trend (although not statistically significant) toward a lower rate of device colonization following receipt of dalbavancin vs. vancomycin (P = 0.07) or saline (P = 0.02). CONCLUSIONS: The demonstrated efficacy of dalbavancin in this animal study suggest that this novel antibiotic may have an important role in the prevention and treatment of device-related infection. PMID- 15780415 TI - Clinical and bacteriological efficacy of 5-day telithromycin in acute maxillary sinusitis: a pooled analysis. AB - OBJECTIVES: To compare the efficacy and tolerability of a 5-day course of telithromycin (800 mg once daily) with a 10-day course of telithromycin or standard comparators (amoxicillin-clavulanate 500/125 mg three times daily or cefuroxime axetil 250 mg twice daily) in patients with acute maxillary sinusitis (AMS). METHODS: Data from three randomised double blind studies were pooled. The studies included patients with clinical symptoms of AMS and sinus X-ray findings of total opacity, air-fluid levels or mucosal thickening. RESULTS: Pooled analysis of results for 5-day telithromycin revealed overall clinical cure rates of 83.6% (383/458 patients) at post-therapy (days 17-24) and 78.9% (330/418 patients) at late post-therapy (days 31-45) in the per-protocol population. Clinical cure rates at post-therapy were equivalent to those observed with 10-day telithromycin (82.5% vs 81.7%) or comparator treatment (80.9% vs 77.4%). Moreover, clinical cure rates exceeded 80% in subgroups of patients of interest, including those with severe infection and those fulfilling more stringent criteria for bacterial AMS. A satisfactory bacteriological outcome was achieved in 87.6% of patients. The 5-day telithromycin regimen was well tolerated. CONCLUSIONS: Telithromycin once daily for 5 days offers effective treatment for AMS and is comparable to 10-day courses of standard treatments. PMID- 15780416 TI - Clinical outcomes of HIV-HCV co-infection in a large cohort of hemophiliac patients. AB - OBJECTIVE: To determine the disease progression of HIV-HCV co-infected hemophiliacs in a large cohort of patients (n = 288) cared for at a single medical institution. PATIENTS AND METHODS: Annual mortality rates for AIDS- and liver-related death were calculated and Kaplan-Meier survival plots were drawn to determine the progression to AIDS and death. RESULTS: Between January 1985 and December 2002, 179 (62.2%) and 195 (67.7%) of these patients had developed AIDS or died, respectively. Overall, AIDS accounted for 128 deaths, which almost entirely (93.7%) occurred prior to the introduction of highly active antiretroviral therapy (HAART) at the end of 1995. A total of 29 patients died of liver failure, most of them (69%) during the years 1991-1996. Since 1997, only five cases of fatal liver failure were reported. Non-HIV-HCV related reasons were responsible for 38 deaths and occurred predominantly (47%) in the years 1997 2002. Starting November 1995, 72 patients were treated with HAART. However, by December 2002, only 52.5% and 83% of all HAART-treated patients had a stable viremia (<400 copies/ml) and a sufficient CD4(+) T-cell count (>200/microl), respectively. CONCLUSION: These data indicate that liver-related mortality peaked in the years 1991-1996, but subsequently tended to decline. Moreover, despite widespread treatment of patients with HAART, a significant proportion of individuals had an unsatisfactory immunological and virological status at the end of 2002. PMID- 15780417 TI - Lopinavir/ritonavir combination and total/HDL cholesterol ratio. AB - OBJECTIVES: To describe the evolution of the lipidic profile among LPV/r treated patients in a 'real life' situation. METHODS: Lipids measurements at LPV/r initiation time and every 3 months, and pharmacological measurements at M3 and M6 were collected retrospectively in 142 patients attending our clinic. Dyslipidaemia was defined as total cholesterol > or =6.2 mmol/l, HDL-cholesterol > or =1 mmol/l, total/HDL-cholesterol ratio > or =6.5 and triglycerides > or =2.3 mmol/l. RESULTS: Eighty-nine percent of patients had previously received a regimen with protease inhibitors, 4% were treatment naive. At baseline, 17% of patients had high total cholesterol, 49% high triglycerides, 63% low HDL cholesterol, 25% a high total/HDL-cholesterol ratio. At M12, the mean HDL cholesterol increase per patient was 21%. Lipids levels significantly increased over the study period, as early as the 3rd month (6th month for ratio) and continuously until the 12th month. Among the patients with available LPV/r plasma determinations at M3, a higher lopinavir residual concentration was observed in those with high triglycerides (6.78 vs 3.02 mg/l, p = 0.05) as, at M6, in those with an elevated ratio (9.19 vs 0.96 mg/l, p = 0.02). CONCLUSIONS: Those results suggest that LPV/r may induce a significant rise in the total/HDL-cholesterol ratio, despite an increase in HDL-cholesterol levels. The association between triglycerides and total/HDL-cholesterol ratio elevated levels and high residual concentrations of lopinavir may also argue for systematic drug monitoring. PMID- 15780418 TI - Fusobacteriosis presenting as community acquired pneumonia. AB - Fusobacterium species are anaerobic Gram-negative bacilli, which colonise the mucus membranes of man and animals and can cause a number of clinical manifestations including Lemierre's disease (postanginal septicaemia), abdominal infection and deep-seated abscesses. The incidence of fusobacterium infections appears to be increasing, and we present three cases of fusobacteriosis who presented with features of community acquired pneumonia (CAP). Cases were treated with benzyl penicillin and metronidazole, co-amoxiclav and metronidazole and amoxicillin and clarithromycin. Since some of the Fusobacterium species are resistant to penicillin and erythromycin, treatment with these antibiotics in cases of fusobacteriosis presenting as CAP may lead to treatment failure. A high index of clinical suspicion is required to recognise this rare cause of CAP. PMID- 15780419 TI - Group B streptococcal meningitis in a patient with horizontal transmission: beware of toothbrushing on Sunday mornings. PMID- 15780420 TI - Antigenic trigger for type 1 reaction in leprosy. AB - Type 1 (reversal or upgrading) reactions occur during or after chemotherapy in around 10% leprosy patients [Int J Lepr Other Mycobact Dis 61 (1993) 8-15]. The cause of this immunological upgrading is incompletely defined, although the approximately 2-fold increased risk of reaction in patients vaccinated with Mycobacterium w suggests that infection by mycobacteria other than Mycobacterium leprae may trigger this phenomenon [Vaccine 13 (1995) 1102-1110]. We report a case of borderline lepromatous leprosy in which we studied the antigenic specificity of peripheral blood mononuclear cells immediately before, and then during, a Type 1 reaction which provides more direct evidence in favor of this hypothesis. PMID- 15780421 TI - Nosocomial ventriculitis due to Roseomonas gilardii complicating subarachnoid haemorrhage. AB - Roseomonas gilardii is a pink-pigmented, non-fermentative, Gram-negative coccobacillus that has been recognized as a rare cause of human infections. We report the first case of ventriculitis caused by R. gilardii in a 54-year-old man with a subarachnoid haemorrhage secondary to a vertebral artery aneurysm; discuss previous reports of this organism as a nosocomial and community-acquired pathogen, laboratory diagnosis, and patient management. PMID- 15780422 TI - Ventriculitis due to a hetero strain of vancomycin intermediate Staphylococcus aureus (hVISA): successful treatment with linezolid in combination with intraventricular vancomycin. AB - A 67-year male presented with relapse 14 days after treatment with vancomycin for a MRSA ventriculitis. CSF samples taken at the time of relapse grew MRSA with a MIC for vancomycin of 4 mg/L by E-test and therapy with linezolid (600 mg bd) and intraventricular vancomycin (20 mg od) was initiated. Using the macrodilution E test, the isolate was found to have sub-populations with a MIC for vancomycin of 8 mg/L and teicoplanin of 12 mg/L and a population analysis profile almost identical to the hVISA strain MU3, indicative of a hVISA strain. Concentrations of vancomycin in the CSF over the period of therapy ranged from 25.6-192.5 mg/L after intraventricular administration and those of linezolid ranged from 3.4-6.7 mg/L after intravenous administration, exceeding the MICs for this isolate. The patient made a successful recovery, with no further episodes of ventriculitis at 1-year follow-up. We report the first case of ventriculitis due to hVISA. It was successfully treated with intrathecal vancomycin and intravenous linezolid. We also believe this to be the first documented case of clinical infection due to hVISA in South Africa. PMID- 15780423 TI - Campylobacter fetus prosthetic hip joint infection: successful management with device retention and review. PMID- 15780424 TI - Chronic hepatotoxicity after long-term antiretroviral treatment including nevirapine. PMID- 15780425 TI - Is Campylobacter jejuni enteritis a weekend disease? PMID- 15780426 TI - Proceedings from the GLP1 Meeting on the Occasion of the 80th Birthday of Prof. Dr. med. Dr. hc (em.) Werner Creutzfeldt. PMID- 15780427 TI - The [pre-] history of the incretin concept. AB - The discoverers of secretin already thought of the existence of a chemical excitant for the internal secretion of the pancreas. Numerous experiments have been performed and published between 1906 and 1935 testing the effect of injected or ingested duodenal ("secretin") extracts on fasting or elevated blood glucose levels of normal or diabetic animals and humans with contradictory results. In 1940, after a series of negative dog experiments performed by an opinion leader, the existence of an incretin was considered questionable and further research stopped for more than 20 years. However, after the development of the radio immunoassay, the incretin-concept has been revived in 1964, showing that significantly more insulin was released after ingestion of glucose than after intravenous injection. The possibility that nerves or one of the known gut hormones were responsible for the incretin effect could be ruled out. In 1970, glucose dependent insulinotropic polypeptide (GIP), and finally, in 1985 glucagon like peptide 1 (GLP-1) and its truncated form GLP-1(7-36) were recognized as true incretins. Thereafter, multiple antidiabetic qualities and the therapeutic perspectives of GLP-1(7-36) and its analogues and mimetics have been demonstratred. PMID- 15780428 TI - The discovery of glucagon-like peptide 1. AB - The discovery of glucagon-like peptide 1 (GLP-1) began more than two decades ago with the observations that anglerfish islet proglucagon messenger RNAs (mRNAs) contained coding sequences for two glucagon-related peptides arranged in tandem. Subsequent analyses revealed that mammalian proglucagon mRNAs encoded a precursor containing the sequence of pancreatic glucagon, intestinal glicentin and two glucagon-related peptides termed GLP-1 and GLP-2. Multidisciplinary approaches were then required to define the structure of biologically active GLP-1 7-36 amide and its role as an incretin, satiety hormone and, most recently, a neuroprotective peptide. This historial perspective outlines the use of traditional recombinant DNA approaches to derive the GLP-1 sequence and highlights the challenges and combination of clinical and basic science approaches required to define the physiology and pathophysiology of bioactive peptides discovered through genomics. PMID- 15780429 TI - Glucagon-related peptide 1 (GLP-1): hormone and neurotransmitter. AB - The interest in glucagon-like petide-1 (GLP-1) and other pre-proglucagon derived peptides has risen almost exponentially since seminal papers in the early 1990s proposed to use GLP-1 agonists as therapeutic agents for treatment of type 2 diabetes. A wealth of interesting studies covering both normal and pathophysiological role of GLP-1 have been published over the last two decades and our understanding of GLP-1 action has widened considerably. In the present review, we have tried to cover our current understanding of GLP-1 actions both as a peripheral hormone and as a central neurotransmitter. From an initial focus on glycaemic control, GLP-1 research has been diverted to study its role in energy homeostasis, neurodegeneration, cognitive functions, anxiety and many more functions. With the upcoming introduction of GLP-1 agonists on the pharmaceutical venue, we have witnessed an outstanding example of how initial ideas from basic science laboratories have paved their way to become a novel therapeutic strategy to fight diabetes. PMID- 15780430 TI - The physiological role of GLP-1 in human: incretin, ileal brake or more? AB - The proglucagon-derived peptide glucagon-like peptide-1 (GLP-1) is an intestinal signal peptide postprandially released from the L cells of the lower gut. Exogenously administered the synthetic hormone exerts a glucose-dependent insulinotropic effect at the pancreatic beta-cells and lowers plasma glucagon by an inhibitory effect against the alpha-cells. It delays gastric emptying by relaxation of the gastric fundus, inhibition of antral contractility, and stimulation of both the tonic and phasic motility of the pyloric sphincter. Enhancement of insulin, suppression of glucagon, and inhibition of gastric emptying are the main determinants controlling glucose homeostasis with GLP-1. Human studies employing the specific GLP-1 receptor antagonist exendin(9-39) show that endogenously released GLP-1 likewise controls fasting plasma glucagon, stimulates insulin, and influences all the motoric mechanisms known to control gastric emptying. Therefore, GLP-1 is discussed as an incretin hormone and as an enterogastrone in man. Synthetic GLP-1 also suppresses gastric acid and pancreatic enzyme secretion. The inhibitory effects on upper gastrointestinal functions are at least partly mediated by vagal-cholinergic inhibition and may involve interactions with vagal afferent pathways and/or circumventricular regions within the CNS. GLP-1 is a candidate humoral mediator of the 'ileal brake' exerting inhibition of upper gastrointestinal function preventing malabsorption and postprandial metabolic disturbances. As human studies indicate a central action of GLP-1 in reduction of food intake, it is uncertain if this is a consequence of induction of satiety or of transduction of visceral aversive stress signals. PMID- 15780431 TI - What do we know about the secretion and degradation of incretin hormones? AB - The incretin hormones, glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1) are secreted from endocrine cells located in the intestinal mucosa, and act to enhance meal-induced insulin secretion. GIP and GLP 1 concentrations in the plasma rise rapidly after food ingestion, and the presence of unabsorbed nutrients in the intestinal lumen is a strong stimulus for their secretion. Nutrients can stimulate release of both hormones by direct contact with the K-cell (GIP) and L-cell (GLP-1), and this may be the most important signal. However, nutrients also stimulate GLP-1 and GIP secretion indirectly via other mechanisms. Incretin hormone secretion can be modulated neurally, with cholinergic muscarinic, beta-adrenergic and peptidergic (gastrin releasing peptide, GRP) fibres generally having positive effects, while secretion is restrained by alpha-adrenergic and somatostatinergic fibres. Hormonal factors may also influence incretin hormone secretion. Somatostatin exerts a local inhibitory effect on the activity of both K- and L-cells via a paracrine mechanism, while, in rodents at least, GIP from the proximal intestine has a stimulatory effect on GLP-1 secretion, possibly mediated via a neural loop involving GRP. Once they have been released, both GLP-1 and GIP are subject to rapid degradation. The ubiquitous enzyme, dipeptidyl peptidase IV (DPP IV) cleaves N-terminally, removing a dipeptide and thereby inactivating both peptides, because the N-terminus is crucial for receptor binding. Subsequently, the peptides may be degraded by other enzymes and extracted in an organ-specific manner. The intact peptides are inactivated during passage across the hepatic bed and further metabolised by the peripheral tissues, while the kidney is important for the final elimination of the metabolites. PMID- 15780432 TI - GIP and GLP-1 as incretin hormones: lessons from single and double incretin receptor knockout mice. AB - Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) are gut-derived incretins secreted in response to nutrient ingestion. Both incretins potentiate glucose-dependent insulin secretion and enhance beta cell mass through regulation of beta-cell proliferation, neogenesis and apoptosis. In contrast, GLP-1, but not GIP, inhibits gastric emptying, glucagon secretion, and food intake. Furthermore, human subjects with Type 2 diabetes exhibit relative resistance to the actions of GIP, but not GLP-1R agonists. The physiological importance of both incretins has been investigated through generation and analysis of incretin receptor knockout mice. Elimination of incretin receptor action in GIPR-/- or GLP-1R-/- mice produces only modest impairment in glucose homeostasis. Similarly, double incretin receptor knockout (DIRKO) mice exhibit normal body weight and normal levels of plasma glucagon and hypoglycemic responses to exogenous insulin. However, glucose-stimulated insulin secretion is significantly decreased following oral but not intraperitoneal glucose challenge in DIRKO mice and the glucose lowering actions of dipeptidyl peptidase-IV (DPP-IV) inhibitors are extinguished in DIRKO mice. Hence, incretin receptor signaling exerts physiologically relevant actions critical for glucose homeostasis, and represents a pharmacologically attractive target for development of agents for the treatment of Type 2 diabetes. PMID- 15780433 TI - Glucagon-like peptide 1 and its derivatives in the treatment of diabetes. AB - Glucagon-like peptide 1 (GLP-1) was discovered as an insulinotropic gut hormone, suggesting a physiological role as an incretin hormone, i.e., being responsible, in part, for the higher insulin secretory response after oral as compared to intravenous glucose administration. This difference, the incretin effect, is partially lost in patients with Type 2 diabetes. The actions of GLP-1 include (a) a stimulation of insulin secretion in a glucose-dependent manner, (b) a suppression of glucagon, (c) a reduction in appetite and food intake, (d) a deceleration of gastric emptying, (e) a stimulation of beta-cell neogenesis, growth and differentiation in animal and tissue culture experiments, and (f) an in vitro inhibition of beta-cell apoptosis induced by different toxins. Intravenous GLP-1 can normalize and subcutaneous GLP-1 can significantly lower plasma glucose in the majority of patients with Type 2 diabetes. GLP-1 itself, however, is inactivated rapidly in vivo and thus does not appear to be useful as a therapeutic agent in the long-term treatment of Type 2 diabetes. Other agents acting on GLP-1 receptors have been found (like exendin-4) or developed as GLP-1 derivatives (like liraglutide or GLP-1/CJC-1131). Clinical trials with exenatide (two injections per day) and liraglutide (one injection per day) have shown reductions in glucose concentrations and HbA1c by more than 1%, associated with moderate weight loss (2-3 kg), but also some nausea and, rarely, vomiting. It is hoped that this new class of drugs interacting with the GLP-1 or other incretin receptors, the so-called "incretin mimetics", will broaden our armamentarium of antidiabetic medications in the nearest future. PMID- 15780434 TI - Glycaemic effects of incretins in Type 1 diabetes mellitus: a concise review, with emphasis on studies in humans. AB - The remission phase of Type 1 diabetes mellitus is associated with substantial recovery of beta-cell function and with marked improvement of endogenous insulin responses to meals in the early months after diagnosis, accompanied by little or no improvement in the insulin response to parenteral glucose, suggesting that the incretin function may be important in glycaemic regulation in this phase of diabetes. Preservation of the insulin response to parenteral glucagon-like peptide-1 (GLP-1), contrasting with lack of stimulation of insulin secretion by the other known incretin gastric inhibitory polypeptide (GIP), prompted studies with exogenous GLP-1 in recent-onset Type 1 diabetes. These studies showed substantial reduction of glycaemic excursions after ingestion of mixed nutrients during intravenous infusion of GLP-1 without administration of insulin, in subjects with a range of endogenous secretion of insulin in response to meals as demonstrated by blood levels of the insulin-connecting peptide (CP). These effects were independent of stimulation of blood levels of CP and were reproduced in volunteers with no endogenous release of CP in response to meals. The glycaemic effects were associated with inhibition of abnormal rises of blood levels of glucagon, and with suppression of endogenous release of human pancreatic polypeptide (HPP), by GLP-1. It was hypothesized that a major component of the glycaemic effect is attributable to the known action of GLP-1 to inhibit gastric emptying and to inhibit glucagon secretion. Studies of the effects of GLP-1 agonists (GLP-1 and exendin-4) given together with established insulin doses before a meal supported the hypothesis. The more prolonged actions of exendin-4 were accompanied by greater and more prolonged reduction of glycaemic effects of ingestion of meals in volunteers with CP-negative Type 1 diabetes mellitus, during intensive insulin therapy, in whom delay of gastric emptying was confirmed by studies of blood levels of acetaminophen ingested with the meals. Side effect-free doses of exendin-4 given together with insulin in volunteers with CP-negative Type 1 diabetes receiving continuing intensive insulin therapy demonstrated the capacity of this combination therapy to normalize blood glucose levels after ingestion of meals that were consistent with the dietary program of the volunteers, without apparent increased risk of hypoglycaemia within a normal between-meals interval. It is suggested that further and more prolonged studies of the use of long-acting GLP-1 agonists as congeners with insulin in Type 1 diabetes mellitus are indicated. PMID- 15780435 TI - Dipeptidyl peptidase IV inhibitors: how do they work as new antidiabetic agents? AB - A number of new approaches to diabetes therapy are currently undergoing clinical trials, including those involving stimulation of the pancreatic beta-cell with the gut-derived insulinotropic hormones (incretins), GIP and GLP-1. The current review focuses on an approach based on the inhibition of dipeptidyl peptidase IV (DP IV), the major enzyme responsible for degrading the incretins in vivo. The rationale for this approach was that blockade of incretin degradation would increase their physiological actions, including the stimulation of insulin secretion and inhibition of gastric emptying. It is now clear that both GIP and GLP-1 also have powerful effects on beta-cell differentation, mitogenesis and survival. By potentiating these pleiotropic actions of the incretins, DP IV inhibition can therefore preserve beta-cell mass and improve secretory function in diabetics. PMID- 15780436 TI - The Rho GTPase activators CNF1 and DNT bacterial toxins have mucosal adjuvant properties. AB - Cytotoxic necrotizing factor 1 (CNF1) from uropathogenic Escherichia coli belongs to a family of factors activating Rho GTPases. We report the in vivo effects of CNF1 in mice co-fed toxin and the soluble protein antigen ovalbumin (OVA). Similar to cholera toxin, CNF1 elicits adjuvanticity anti-OVA responses, both systemic and mucosal. In contrast, the catalytic inactive mutant CNF1-C866S demonstrated no effects. Using dermonecrotic toxin (DNT), a closely related Rho activating toxin from Bordetella, we discovered that the adjuvant property is within the DNT catalytic domain. Manipulation of Rho proteins thus provides a possible new approach for the development of effective mucosal immunoadjuvants. PMID- 15780437 TI - Protective immunity against Mycobacterium bovis induced by vaccination with Rv3109c--a member of the esat-6 gene family. AB - In a number of clinical studies the current TB vaccine, Mycobacterium bovis bacille Calmette-Guerin (BCG), has provided little or no protection against pulmonary tuberculosis in cattle and man. A new generation of vaccines is therefore required to replace or supplement BCG. Safety concerns surrounding a number of strategies make protein subunits an attractive approach. Moreover, novel prime-boost strategies based on primary immunisations with BCG are not only showing promise but also present a clear strategy for testing new TB vaccines in clinical studies. We report the evaluation of six protein vaccine candidates for their ability to induce protective immunity in a murine virulent M. bovis challenge model. One protein (Rv3019c) induced reproducibly significant protection in the spleen and lungs approaching that induced by BCG. Detailed analysis of antigen-specific T cell responses revealed that despite robust responses in the spleen and lungs of vaccinated mice, there was no correlation between these responses and the protective efficacy of the vaccine. Significantly, Rv3019c also stimulated IFN-gamma responses in PBMC from BCG vaccinated cattle, indicating its potential for use in a heterologous prime-boost strategy in conjunction with BCG in the target species. PMID- 15780438 TI - Role of IgA in the defense against respiratory infections IgA deficient mice exhibited increased susceptibility to intranasal infection with Mycobacterium bovis BCG. AB - IgA is the predominant Ig isotype in mucosal tissue and is believed to be involved in defense against viral and bacterial infections at these sites. Here, we examined the role of IgA in the protection against intranasal (i.n.) infection with Mycobacterium bovis Bacillus Calmette-Guerin (BCG). IgA deficient (IgA-/-) mice and wild type non-targeted littermate (IgA+/+) mice were immunized by i.n. route with the mycobacterium surface antigen PstS-1 formulated with cholera toxin (CT). Our data showed that IgA-/- mice were more susceptible to BCG infection compared to IgA+/+ mice, as revealed by the higher bacterial loads in the lungs and bronchoalveolar lavage (BAL). Analysis of the Ig levels and the antibody responses to PstS-1 showed that IgA-/- mice had no detectable IgA either in the saliva or in the BAL. However, these mice displayed higher levels of total and specific IgM than IgA+/+ mice in both mucosal fluids. More importantly, analysis of the cytokine responses revealed a reduction in the IFN-gamma and TNF-alpha production in the lungs of IgA-/- compared to IgA+/+ mice. Altogether, our results suggest that IgA may play a role in protection against mycobacterial infections in the respiratory tract by blocking the pathogen entrance and/or by modulating the pro-inflammatory responses. PMID- 15780439 TI - Evaluation of the xerovac process for the preparation of heat tolerant contagious bovine pleuropneumonia (CBPP) vaccine. AB - The study was conducted with the aim of evaluating the xerovac process as a method for preparing contagious bovine pleuropneumonia (CBPP) vaccine with increased heat resistance. The thermo-protective effects of various concentrations of trehalose in mycoplasma growth medium, various concentrations of trehalose in the dehydration stabilizer and the importance of some divalent cations were assessed. The results obtained indicate that a rapid dehydration of CBPP vaccine following the xerovac method and in an excipient composed of a high concentration of trehalose, renders the product more heat tolerant than a similar vaccine prepared using a regular or an extended freeze drying regime. It was also demonstrated that the addition of chitosan as a mycoplasma precipitating agent conferred additional heat resistance to the vaccine. It is suggested that the application of the xerovac process in the dehydration of CBPP vaccine offers the advantages of a faster, cheaper and easier process over the conventional dehydration methods like freeze drying. PMID- 15780440 TI - Evaluation of an intracellular-activated promoter for the generation of live Salmonella recombinant vaccines. AB - Salmonella enterica serovar Typhimurium is a versatile host organism for the generation of recombinant live vaccines for mucosal immunization. We investigated the performance of an intracellular-activated promoter for expression of heterologous antigens by Salmonella carrier strains. We observed that intracellular expressed antigens were more efficient in elicitation of humoral and cellular immune responses than constitutively expressed antigens. Furthermore, the intracellular-activated promoter was more efficient in combination with a carrier strain deficient in SPI2 gene sseC than with the conventional aroA strain. We propose that intracellular-activated promoters will be useful tools for construction of efficient recombinant live vaccines. PMID- 15780441 TI - Potent enhancement of cellular and humoral immune responses against recombinant hepatitis B antigens using AS02A adjuvant in healthy adults. AB - Recombinant subunit protein vaccines generally elicit good humoral immune responses, weak helper T cell responses and no cytotoxic T cell responses. Certain adjuvants are known to enhance humoral and cellular immune responses. This study evaluated the humoral, CD4+ T helper and CTL responses induced by the recombinant SL* protein adjuvanted with AS02A in comparison with non-adjuvanted SL* in PBS in two groups of 15 healthy adult volunteers. The AS02A adjuvant contains monophosphoryl lipid A (MPL), QS21 and an oil in water emulsion. The adjuvanted vaccine induced fast and vigorous humoral and helper T cell responses of the Th1 type. Using a pool of overlapping 20mer peptides a cytotoxic response was detected in 6 out of 14 HLA-A2-positive (+) and HLA-A2-negative (-) recipients of the adjuvanted vaccine. All HLA-A2-positive subjects in the adjuvanted group and up to 30% of the subjects in the SL* PBS group displayed a CTL response against selected HLA-A2-restricted CD8+ T cell epitopes. The non adjuvanted vaccine induced a very weak antibody response and no helper T cell responses. Local and general reactions were more frequently reported by AS02A recipients than in the non-adjuvanted group but the safety profile was considered acceptable. AS02A can be considered as a useful adjuvant that strongly enhances the cellular and humoral responses of subunit protein vaccines. PMID- 15780442 TI - Immunogenicity and safety of a combined hepatitis A and B vaccine administered concomitantly with either a measles-mumps-rubella or a diphtheria-tetanus acellular pertussis-inactivated poliomyelitis vaccine mixed with a Haemophilus influenzae type b conjugate vaccine in infants aged 12-18 months. AB - Two studies were undertaken to investigate the concomitant administration of combined hepatitis A/B vaccine with a diphtheria-tetanus-acellular pertussis inactivated poliomyelitis vaccine mixed with Haemophilus influenzae vaccine (DTPa IPV/Hib), or with a measles-mumps-rubella vaccine (MMR), during the second year of life. On completion of the vaccination course, all subjects were seropositive or seroprotected against all antigens except for one subject who was seronegative for anti-PT. Seropositivity and seroprotection rates for all other antibodies were comparable to reference values for each vaccine component, indicating that the immunogenicity of MMR, DTPa-IPV/Hib and combined hepatitis A/B vaccines is not impaired by co-administration. All vaccines were well tolerated. PMID- 15780443 TI - Pneumococcal conjugate vaccination does not induce a persisting mucosal IgA response in children with recurrent acute otitis media. AB - AIM: In a prospective controlled study in young children with a history of recurrent acute otitis media, we analyzed the salivary IgA and IgG antibody titers upon vaccination with a 7-valent pneumococcal conjugate vaccine (PCV) given once or twice, followed by a 23-valent polysaccharide booster vaccination. METHODS: Salivary IgA and IgG antibody concentrations to vaccine serotype 6B, 14, 18C and 19F were measured by enzyme immunoassay in 38 samples of children vaccinated with PCV and 45 control samples. In the PCV group, 12 samples were taken prior to vaccination, 12 samples 4 weeks after the polysaccharide booster (8 months after the first conjugate vaccination) and 14 samples 7 months after the last vaccination (14 months after the first conjugate vaccination). In the control group 15 children were sampled at each of these three time points. RESULTS: We observed an increase in salivary IgG antibody concentrations against serotype 6B, 14, and 18C 14 months after the primary vaccination in children vaccinated with PCV twice, although this was significant for serotype 14 only. There was no increase in salivary IgG antibody in children vaccinate with PCV once nor in control children. IgA antibody titers increased significantly after 8 and after 14 months in both the pneumococcal vaccine recipients and the controls. However, the observed increase in mean antibody titers was significantly higher in control children compared to the PCV group. CONCLUSION: We suggest that repeated pneumococcal conjugate vaccination is necessary to induce an increase in salivary IgG antibodies and effectuate clearance of S. pneumoniae from the nasopharyngeal mucosa of children with recurrent acute otitis media. We hypothesize that the increase in salivary IgA is caused by the local boosting of the mucosal immune response by carriage and recurrent infections, which occurs less often in the PCV group compared to the control children. PMID- 15780444 TI - Oral administration of second-generation immunomodulatory oligonucleotides induces mucosal Th1 immune responses and adjuvant activity. AB - CpG DNA induces potent Th1 immune responses through Toll-like receptor 9. In the present study, we used oligonucleotides consisting of a novel 3'-3'-linked structure and synthetic stimulatory motifs, referred as second-generation immunomodulatory oligonucleotides (IMOs). The stimulatory motifs included: CpR, YpG, or R'pG (R = 2'-deoxy-7-deazaguanosine, Y = 2'-deoxy-5-hydroxy-cytidine, and R' = 1-[2'-deoxy-beta-d-ribofuranosyl]-2-oxo-7-deaza-8-methyl-purine). We evaluated the stability of orally administered IMOs in the gastrointestinal (GI) environment and their ability to induce mucosal immune responses in mice, and compared these characteristics with those of a conventional CpG DNA. The IMOs were significantly more stable than CpG DNA following oral administration, and IMOs induced stronger local and systemic immune responses as determined by MIP 1beta, MCP-1, IP-10, and IL-12 production. Mice orally immunized with ovalbumin (OVA) and IMO had higher levels of IgG2a antibodies in serum and IgA antibodies in intestinal mucosa than did mice immunized with OVA and CpG DNA. These studies demonstrate that IMOs are more stable than CpG DNA in the GI tract and can induce more potent mucosal Th1 adjuvant responses. IMOs may prove to be effective oral adjuvants, able to promote strong systemic and mucosal immune responses to oral vaccines and antigens for therapeutic and prophylactic applications. PMID- 15780445 TI - Effect of sequence variation in meningococcal PorA outer membrane protein on the effectiveness of a hexavalent PorA outer membrane vesicle vaccine in toddlers and school children. AB - Though meningococcal conjugate vaccines are effective against serogroup C, there is currently no vaccine solution for serogroup B disease. PorA outer membrane protein (OMP) is a potential serogroup B vaccine candidate. A hexavalent PorA outer membrane vesicle (OMV) vaccine has been evaluated in phase I and II trials with promising results. However, considerable sequence variation occurs in the variable regions (VRs) encoding these serosubtypes. By using five wild type P1.19,15 variant strains we examined the serum bactericidal antibody (SBA) titres from sera collected from toddlers and school children pre- and post-vaccination. The numbers of subjects with SBA titres of <4, 4 and > or = 8 varied greatly between the different strains. This was also reflected when > or = 4-fold rises in SBA titres were examined. This finding in sera from toddlers and school children may have implications for PorA based vaccines. PMID- 15780446 TI - Safety characterization of HeLa-based cell substrates used in the manufacture of a recombinant adeno-associated virus-HIV vaccine. AB - The use of transformed cell substrates for prophylactic vaccine manufacturing is widely debated. Extensive characterization is required to address the suitability of neoplastic cell substrates for vaccine manufacture. The HeLa-based cell substrate used in the manufacture of a prophylactic rAAV-HIV vaccine, AAV2-gagPR delta RT (tgAAC09) was tested in vivo for its tumor-forming potential, the oncogenic potential of its high molecular weight DNA and the potential presence of occult oncogenic adventitious agents. This data from these in vivo studies, in conjunction with prion gene and protein characterization, cell and viral clearance studies and quantity of residual host-cell DNA levels in the purified tgAAC09 vaccine, were used to establish what we believe to be an acceptable safety profile for the vaccine manufacturing process. The tumor-producing dose in 50% of the animals was consistent with that in a published report from FDA staff for HeLa cells. High molecular weight cellular DNA was not oncogenic and no occult oncogenic agents were detected by testing in nude mice and newborn rodent models, respectively. Endogenous prion protein was also normal and genomic sequence analysis detected no mutations associated with increased risk of prion disease. In addition, the purification process used to produce this vaccine candidate removed all detectable cells (clearance of greater than 22 log10), viral clearance study showed 6-17 log10 clearance of three model viruses and host cell DNA in the bulk product was less than 100pg host-cell DNA per dose of 3 x 10(11) DNase resistant particles (DRP) of the vaccine. Taken together, the data from the in vivo and in vitro tests that were performed to characterize the HeLa based producer cell line (T3B12-5B) and HeLa S3 cells support the use of these cells as substrates for the manufacture of a purified rAAV-HIV vaccine candidate. The data also supports the ability of the process, employing the HeLa cell substrate, used to manufacture the rAAV-HIV vaccine to produce a product as free of adventitious agents as current testing procedures can document. Safety of the rAAV-HIV vaccine is currently being assessed in a Phase I clinical trial. PMID- 15780447 TI - CD4+ T cell epitope affinity to MHC II influences the magnitude of CTL responses elicited by DNA epitope vaccines. AB - Immunization with naked plasmid DNA elicits strong cell-mediated immune responses. In the present study, we examine strategies to enhance epitope specific cytotoxic T lymphocyte (CTL) responses using DNA constructs, expressing a minimal class I epitope of the gp120 of HIV-IIIB. Here, we evaluate the effect of CD4+ T cell (T(H)) epitope affinity for the MHC II molecule on the immunogenicity of our DNA vaccines. Our data indicate that a low-affinity T(H) epitope decreased the magnitude of the CTL responses. In addition, we observed decreased numbers of epitope-specific T helper cells and CTLs, as well as diminished cytokine secretion and proliferative responses. Thus, the immunogenicity of a DNA epitope vaccine can be modulated by altering the affinity of the T(H) epitope. PMID- 15780448 TI - Analysis of the immune response against mixotope peptide libraries from a main antigenic site of foot-and-mouth disease virus. AB - The design of vaccines for RNA viral diseases is complicated by the high genetic variability of the viruses, which favors the selection of escape mutants. A case in point is foot-and-mouth disease virus (FMDV), for which only limited protection has been observed in vaccination with single peptides. We have explored the potential of immunogens of higher sequence diversity, covering a broad range of field or culture-induced mutations at the immunodominant site A of FMDV, serotype C. Four mixotope-type peptide libraries, containing ca. 3 x 10(3) or ca. 3 x 10(5) peptides each, in either linear or cyclic form, and combining most significant mutations found or induced at site A have been synthesized and used to immunize guinea-pigs. Substantial levels of serum conversion have been observed for all four mixotope libraries, as well as for single peptides, linear or cyclic, corresponding to the consensus site A sequence. The specificity and neutralizing ability of the anti-mixotope and -peptide antibodies have been evaluated by direct ELISA and by plaque reduction and micro-neutralization assays, respectively. Challenge experiments with an infectious, guinea-pig adapted FMDV strain, have shown higher protection rates in animals immunized with the cyclic versions, either in single sequence or in combinatorial mixotope form. PMID- 15780449 TI - Vaccination with recombinant HBsAg-HBIG complex in healthy adults. AB - A therapeutic vaccine for viral hepatitis B composed of yeast-derived recombinant HBsAg complexed to human anti-HBs immunoglobulin (yeast-derived-immunogenic complex, YIC) with alum as the adjuvant was evaluated for safety. In stage 1, 22 healthy Chinese adult volunteers were vaccinated with three doses of 30 microg, 60 microg or 90 microg of HBsAg in YIC at 4-week intervals. In stage 2, nine volunteers received 90 microg of HBsAg in YIC for six injections. All immunizations were well tolerated. Renal, liver function and other blood chemistry tests remained within normal range. All recipients developed serum anti HBs, the highest being 1000 mIU/ml, and the subtypes of anti-HBs were IgG1 and IgG3. The serum levels of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) were increased, while no significant increase was observed in interleukin-4 (IL 4), interleukin-6 (IL-6), interleukin-10 (IL-10) or tumor necrosis factor-alpha (TNF-alpha). These results indicate that this complex is safe and can induce a potent anti-HBs response. PMID- 15780450 TI - The immunogenicity-enhancing effect of emulsion vaccine adjuvants is independent of the dispersion type and antigen release rate--a revisit of the role of the hydrophile-lipophile balance (HLB) value. AB - Effective antigen delivery is one of the most important issues in vaccine development. It has been suggested that adjuvant action results from a depot effect by prolonging the duration of the interaction between antigen and cells, and thus is related to the antigen-releasing properties of emulsion adjuvants. The objective of this study was to investigate the effect of the dispersion properties of emulsion-type vaccine adjuvants on the immune response with the aim of optimizing vaccine adjuvant formulation. Emulsion-type adjuvants with various dispersion properties of either the oil-in-water or water-in-oil type were prepared using emulsifiers with various hydrophilic-hydrophobic balance (HLB) values. The physicochemical properties of the emulsions, including the conductivity and viscosity, and antigen release rates were then determined. Cell death induced by the vaccine adjuvants was examined in EL4 cells by Annexin V/propidium iodide (PI) staining and flow cytometric analysis. Mice were immunized with or without the adjuvants and the immunogenicity-enhancing effect of the adjuvants determined by measuring antibody production using an enzyme linked immunosorbent assay. The conductivity, viscosity, and antigen release rates varied widely among emulsions containing emulsifiers with different HLB values. However, the magnitude of the antigen-specific antibody response was similar in most emulsions adjuvants containing Spans or Tweens. L121-adjuvant, the control adjuvant inducing the strongest apoptosis in vitro, was shown to stimulate the highest antibody response in vivo. The results obtained in this study indicate that the immunogenicity-enhancing effect of emulsion adjuvants is independent of the dispersion type and the antigen release rate of the vaccine delivery system. PMID- 15780451 TI - BCG vaccine-associated suppurative lymphadenitis. AB - Clusters of BCG vaccine-associated suppurative lymphadenitis (BCG-ASLD) have been reported in many countries. We have observed an increase in BCG-ASLD following the change from the percutaneous Evans vaccine to the intradermal BCG Statens Serum Institute (SSI) vaccine. PMID- 15780452 TI - Efficacy of the live attenuated Francisella tularensis vaccine (LVS) in a murine model of disease. AB - A live attenuated vaccine Francisella tularensis live vaccine strain (LVS), that confers protection against tularemia infection in a number of animal models including man was developed during the 1960s in the US. In this study, we have established the median lethal dose (MLD) after intraperitoneal (i.p.) or intravenous (i.v.) delivery of NDBR Lot 4 F. tularensis LVS to be 4 cfu and 2.24 x 10(4) cfu, respectively, in BALB/c mice and less than 1 cfu and 1.29 x 10(4) cfu, respectively, in C57BL/6 mice. When delivered subcutaneously, the MLD for F. tularensis LVS was greater then 1 x 10(8) cfu in both strains of mouse. Using mouse models of systemic tularemia infection it was demonstrated that F. tularensis LVS immunised BALB/c mice were fully protected after challenge with approximately 1000 MLD of a strain of F. tularensis subsp. tularensis or a strain of F. tularensis subsp. holarctica. Under similar challenge conditions, protection in C57BL/6 mice was only evident against a subsp. holarctica strain. In BALB/c mice, protection against a subsp. holarctica strain was achieved 4 days after F. tularensis LVS immunisation whereas protection against a subsp. tularensis strain was only evident 14 days after F. tularensis LVS immunisation. PMID- 15780453 TI - Real-life spatial skills, handedness, and family history of handedness. AB - According to , pronounced left hemisphere lateralization for language abilities in women, as in female absolute right-handers, limits their right hemisphere capacity and spatial abilities. This study examines the degree of handedness and the family history of non-right-handedness with respect to real-life spatial abilities in women. Twenty-four women had, first, to learn a new route and, second, to orient themselves within a labyrinth. In the former task, the number of errors and completion time were evaluated; in the latter task, degree of error for orienting was recorded. The results show that, contrary to Annett's prediction, right-handers with and without a family history of non-right handedness did not differ on these measures. In addition, and unexpectedly, absolute right-handers were found to surpass non-absolute ones in the spatial orientation task. These findings do not support Annett's hypothesis and are discussed in relation to functional cerebral organization. PMID- 15780454 TI - Semantic and syntactic aspects of the mass/count distinction: a case study of semantic dementia. AB - Semantic and syntactic contributions to processing of mass and count nouns were assessed by examining the performance of a patient suffering from a pure semantic deficit. Semantic and syntactic processing was evaluated on grammaticality judgement and sentence-picture matching tasks, respectively, where each task involved mass and count readings of metonymic nouns. While the patient did not show impaired performance on the grammaticality judgment task, he manifested difficulties in making mass/count distinctions in the sentence-picture matching task. It is thus argued that while distributionally the mass/count distinction may be established on a purely syntactic basis, cognitive processing of mass/count information requires both intact syntactic and semantic knowledge. PMID- 15780455 TI - Canceling out both the real and the spectral lines. AB - Neglect patients typically show motor perseveration while canceling targets on the ipsilesional side. This behavior can be influenced by the presence vs. absence of targets on the (neglected) contralesional side (). As alternative explanations, the authors proposed (i) directional hypokinesia--the patient cannot perform reaching movements towards detected left-sided targets, and thus carries on canceling on the right side, and (ii) allochiria--the patient misperceives left-sided targets as located on the right side, and cancels them there. We report here data from a patient (EZ) that might confirm the second hypothesis. EZ was presented with 19 displays in which the number and position of cancellation targets on both sides were varied systematically. EZ showed motor perseveration while canceling, but this tendency did not vary across conditions. Interestingly though, EZ also drew cancellation marks in the empty space between the ipsilesional targets, and this phenomenon was significantly more intense when there were more targets on the neglected side. As EZ's comments suggested, such a behavior might reflect the attempt to cancel out delusional targets. Our speculation is that those objects were generated by allochiria. PMID- 15780456 TI - Early Huntington's disease affects movements in transformed sensorimotor mappings. AB - This study examined the effect of transformed visual feedback on movement control in Huntington's disease (HD). Patients in the early stages of HD and controls performed aiming movements towards peripheral targets on a digitizing tablet and emphasizing precision. In a baseline condition, HD patients were slower but showed few precision problems in aiming. When visual feedback was inverted in both vertical and horizontal axes, patients showed problems in initial and terminal phases of movement where feedback is most critical. When visual feedback was inverted along a single axis as in a mirror-inversion, HD patients showed large deviations and over-corrections before adaptation. Adaptation was similar in both groups. These results suggest that HD impairs on-line error correction in novel movements. PMID- 15780457 TI - Emotion-based learning and central executive resources: an investigation of intuition and the Iowa Gambling Task. AB - The role of emotion in complex decision-making can be assessed on the Iowa Gambling Task (IGT), a widely used neuropsychological measure that may tap a different aspect of executive function than that assessed by conventional measures. Most notably, the 'feeling' about which decks are good or bad, often described in relation to IGT performance, seems reminiscent of decision-making based on intuition-linked to a long history of research in decision-making contrasting the 'intuition' versus 'reasoning' styles of problem solving. To test the claim that the performance on the IGT relies more on emotion-based learning than conventional executive resources for normal performance, a group of participants completed the IGT simultaneously with one of two secondary-tasks, one of which (random number generation) is known to load executive resources. A third group performed the IGT with no secondary-task. If performance on the IGT requires the properties associated with intuitive operations, then participants should either show no disruption when completing a secondary-task, or at least show no selective disruption on a secondary-task that loads for executive function. The rate of learning in the three groups was not significantly different. This suggests that the sorts of cognitive resources loaded by traditional executive tasks such as random number generation do not overlap, in the cognitive architecture, with the emotion-based learning skills that are required for Iowa Gambling Task performance. The findings of the present study are also consistent with a previous claim of the Iowa group that emotion-based learning and working memory resources are doubly dissociable. PMID- 15780458 TI - Past experience influences object representation in working memory. AB - The nature of object representation in working memory is vital to establishing the capacity of working memory, which in turn shapes the limits of visual cognition and awareness. Although current theories discuss whether representations in working memory are feature-based or object-based, no theory has considered the role of past experience. However, work with humans and non human primates suggests that once participants learn which features are important for category membership, these diagnostic features become more salient than non diagnostic features in long-term memory and object recognition. Critically, the brain areas involved in this diagnosticity effect are also recruited during working memory tasks. We report two experiments testing whether a diagnosticity effect exists in working memory; and whether it is present when visual information is encoded into working memory, or if it is the result of maintenance within working memory. Results showed a diagnosticity effect which was present at encoding. Maintenance did not influence the nature of object representation in working memory. These findings show that the meaning we glean from our past experience has a profound influence on the nature of object representation in working memory. PMID- 15780459 TI - Action control: independent effects of memory and monocular viewing on reaching accuracy. AB - Evidence suggests that perceptual networks in the ventral visual pathway are necessary for action control when targets are viewed with only one eye, or when the target must be stored in memory. We tested whether memory-linked (i.e., open loop versus memory-guided actions) and monocular-linked effects (i.e., binocular versus monocular actions) on action arise from a common mechanism as suggested by evidence from neuropsychology and psychophysics. Participants viewed targets with either one eye or two (vision: monocular versus binocular) and then reached to touch targets in open-loop and memory-guided conditions (condition: open-loop versus 0, 500, 1000, and 1500 ms delays). Results showed that memory-linked and monocular-linked increases in radial and variable movement error were additive (i.e., main effects of vision and condition, but no interaction). This suggests that the two effects on visuomotor control arise from separate mechanisms, in contrast to evidence from psychophysics and neuropsychology suggesting a common underlying mechanism. PMID- 15780460 TI - Appearance of symmetry, beauty, and health in human faces. AB - Symmetry is an important concept in biology, being related to mate selection strategies, health, and survival of species. In human faces, the relevance of left-right symmetry to attractiveness and health is not well understood. We compared the appearance of facial attractiveness, health, and symmetry in three separate experiments. Participants inspected front views of faces on the computer screen and judged them on a 5-point scale according to their attractiveness in Experiment 1, health in Experiment 2, and symmetry in Experiment 3. We found that symmetry and attractiveness were not strongly related in faces of women or men while health and symmetry were related. There was a significant difference between attractiveness and symmetry judgments but not between health and symmetry judgments. Moreover, there was a significant difference between attractiveness and health. Facial symmetry may be critical for the appearance of health but it does not seem to be critical for the appearance of attractiveness, not surprisingly perhaps because human faces together with the human brain have been shaped by adaptive evolution to be naturally asymmetrical. PMID- 15780461 TI - Spatial awareness, alertness, and ADHD: the re-emergence of unilateral neglect with time-on-task. AB - Studies examining a relationship between attention deficit hyperactivity disorder (ADHD) and relative visual inattention towards left space have produced inconsistent results. Here, based on previous studies with adult neurological patients who show very severe inattention to the left, we examine whether any spatial bias in ADHD may be modulated by changes in alertness levels. In a single case, we found that inattention to the left--that was intermittently apparent- dramatically increased as a function of time-on-task. No significant changes were observed on the right. The implications for clinical assessment and research design are discussed. PMID- 15780462 TI - Hippocampal and amygdalar involvement in discriminatory place learning. AB - A conflict task was developed that simultaneously examines place aversion learning and fear-motivated context discrimination. The task superimposed Pavlovian discriminative fear conditioning on an appetitively motivated instrumental response (alternation). Rats were trained to alternate along a high walled, diamond-shaped runway between two chambers for food. On half of the trials, a tone CS signaled the fact that a fixed section at the apex of the runway was electrified. Both the tone and the shock were turned on at the beginning of, and remained on for the duration of, each tone trial. A new trial was initiated at the time the animal entered the subsequent food chamber. Therefore, during a tone trial, in order to attain additional food reinforcement, the animal had to cross over the electrified region at the runway apex. Behavioral performance of rats with small lesions of the amygdala or dorsal hippocampus (DH) was compared with that of sham-operated controls. All groups displayed significant discriminative responding, hesitating more on tone trials while in areas of the runway adjacent to the shock region. Animals with lesions of the DH were similar to controls with respect to the tone-mediated discrimination, yet were delayed in the initial expression of a location-specific fear response. Conversely, amygdala lesions did not affect place learning; however, these animals were impaired in their suppression of the fear response following repeated unpaired trials. PMID- 15780463 TI - Time course of behavioral changes following basal forebrain cholinergic damage in rats: Environmental enrichment as a therapeutic intervention. AB - The present experiment was designed to study changes in behavior following immunolesioning of the basal forebrain cholinergic system. Rats were lesioned at 3 months of age by injection of the 192 IgG-saporin immunotoxin into the medial septum area and the nucleus basalis magnocellularis, and then tested at different times after surgery (from days 7-500) on a range of behavioral tests, administered in the following order: a nonmatching-to-position task in a T-maze, an object-recognition task, an object-location task, and an open-field activity test. The results revealed a two-way interaction between post-lesion behavioral testing time and memory demands. In the nonmatching-to-position task, memory deficits appeared quite rapidly after surgery, i.e. at a post-lesion time as short as 1 month. In the object-recognition test, memory impairments appeared only when rats were tested at late post-lesion times (starting at 15 months), whereas in the object-location task deficits were apparent at early post-lesion times (starting from 2 months). Taking the post-operative time into account, one can hypothesize that at the shortest post-lesion times, behavioral deficits are due to pure cholinergic depletion, while as the post-lesion time increases, one can speculate the occurrence of a non-cholinergic system decompensation process and/or a gradual degeneration process affecting other neuronal systems that may contribute to mnemonic impairments. Interestingly, when middle-aged rats were housed in an enriched environment, 192 IgG-saporin-lesioned rats performed better than standard-lesioned rats on both the nonmatching-to-position and the object recognition tests. Environment enrichment had significant beneficial effects in 192 IgG-saporin-lesioned rats, suggesting that lesioned rats at late post-lesion times (over 1 year) still have appreciable cognitive plasticity. PMID- 15780464 TI - The primate amygdala: Neuronal representations of the viscosity, fat texture, temperature, grittiness and taste of foods. AB - The primate amygdala is implicated in the control of behavioral responses to foods and in stimulus-reinforcement learning, but only its taste representation of oral stimuli has been investigated previously. Of 1416 macaque amygdala neurons recorded, 44 (3.1%) responded to oral stimuli. Of the 44 orally responsive neurons, 17 (39%) represent the viscosity of oral stimuli, tested using carboxymethyl-cellulose in the range 1-10,000 cP. Two neurons (5%) responded to fat in the mouth by encoding its texture (shown by the responses of these neurons to a range of fats, and also to non-fat oils such as silicone oil ((Si(CH(3))(2)O)(n)) and mineral oil (pure hydrocarbon), but no or small responses to the cellulose viscosity series or to the fatty acids linoleic acid and lauric acid). Of the 44 neurons, three (7%) responded to gritty texture (produced by microspheres suspended in cellulose). Eighteen neurons (41%) responded to the temperature of liquid in the mouth. Some amygdala neurons responded to capsaicin, and some to fatty acids (but not to fats in the mouth). Some amygdala neurons respond to taste, texture and temperature unimodally, but others combine these inputs. These results provide fundamental evidence about the information channels used to represent the texture and flavor of food in a part of the brain important in appetitive responses to food and in learning associations to reinforcing oral stimuli, and are relevant to understanding the physiological and pathophysiological processes related to food intake, food selection, and the effects of variety of food texture in combination with taste and other inputs on food intake. PMID- 15780465 TI - Antinociceptive effects of choline against acute and inflammatory pain. AB - We used the hot plate test and the formalin test to evaluate the antinociception of choline after i.c.v. or i.v. administration. The analgesic mechanism of choline was also studied. The response latency of mice was significantly prolonged in the hot plate test after choline (90-120 mug/animals) i.c.v. administration in a dose-dependent manner. Pretreatment with methyllycaconitine citrate (MLA), alpha-bungarotoxin, or atropine blocked the antinociception of choline in the hot plate test. In contrast, mecamylamine and naloxone had no effect. No antinociceptive action of choline was found in the hot plate test, but it did have an effect in the late phase of the formalin test after i.v. administration. The effect of choline on anti-inflammatory pain was blocked by MLA, but not by mecamylamine, naloxone and atropine, which is indicative of the involvement of alpha7 receptors in peripheral sites. When choline (2 mg/kg) was coadministered with aspirin (9.4 mg/kg), the licking/biting times in the late phase significantly decreased, although no effects were shown when these doses of drugs were used alone. Similarly, coadministration of choline (2 mg/kg) with morphine (0.165 mg/kg) significantly increased the antinociception of morphine in the late phase, but had no effect in the early phase. These results demonstrate that activation of alpha7 nicotinic receptors by choline elicits antinociceptive effects both in an acute thermal pain model and in an inflammatory pain model. Choline holds promise for development as a non-addictive analgesic drug and in reducing the regular dose of aspirin or morphine in inflammatory pain. PMID- 15780466 TI - Role of hippocampal M2 muscarinic receptors in the estrogen-induced enhancement of working memory. AB - We have previously demonstrated that acetylcholine, acting at M2 muscarinic receptors, mediates the estradiol-induced increase in hippocampal N-methyl-d aspartate receptor binding and the associated enhancement in working memory. The goal of present experiment was to investigate the role of hippocampal M2 receptors in the behavioral aspects of these effects. Ovariectomized rats were trained to locate a hidden escape platform on a matching-to-place version of the water maze in which the platform was moved to a new location for each session of four daily trials. Following 18 days of training, rats were randomly assigned to receive one of the following treatments: 1) injections of oil vehicle delivered 72 and 48 h before testing and continuous delivery of vehicle into the dorsal hippocampus via bilateral cannulae implants connected to osmotic minipumps; 2) injections of estradiol benzoate (EB) delivered 72 and 48 h before testing and continuous delivery of vehicle into the hippocampus; 3) injections of EB delivered 72 and 48 h before testing and continuous delivery of the M2 muscarinic receptor antagonist, AFDX 116, into the hippocampus; and 4) injections of EB delivered 72 and 48 h before testing and continuous delivery of AFDX 116 into a control site in the cortex. Chronic administration of AFDX 116 into the hippocampus, but not the cortex, significantly attenuated an estrogen-induced enhancement in performance on a working memory task in the water maze as indicated by increased latency and increased path length to locate an escape platform during a test trial when a 90 min delay was imposed between the first and second trials. These results indicate that acetylcholine acts at M2 muscarinic receptors located in the hippocampus to mediate the positive effects exerted by estrogen on working memory. PMID- 15780467 TI - Brain creatine functions to attenuate acute stress responses through GABAnergic system in chicks. AB - The involvement of brain creatine in the adaptation to acute stress responses was investigated in chicks. In experiment 1, brain creatine content of chicks exposed to social separation stress was significantly increased compared with control chicks. The effects of i.c.v. injection of creatine (2 mug) on vocalizations, spontaneous activity and plasma corticosterone concentration in chicks under social separation stress were investigated in experiment 2. All measurements were attenuated by the i.c.v. injection of creatine compared with the controls under separation stress. Creatine also significantly decreased the active posture, but increased the motionless eye-opened posture, compared with the control. To clarify the relationship between creatine function and GABA receptors, the i.c.v. co-injection of creatine with picrotoxin, a GABA-A receptor antagonist, or CGP54626, a GABA-B receptor antagonist, was investigated in experiments 3 and 4. The effects of creatine on vocalizations and spontaneous activity were attenuated by co-injection of picrotoxin. In this case, active postures decreased by creatine were recovered by co-injection with picrotoxin. However, these effects were not obtained with CGP54626. The results suggest that central creatine functions within the CNS to attenuate the acute stress response by acting through GABA-A receptors in chicks. PMID- 15780469 TI - Tenascin-C regulates proliferation and migration of cultured astrocytes in a scratch wound assay. AB - Tenascin-C (TNC), an extracellular matrix glycoprotein, is involved in tissue morphogenesis like embryogenesis, wound healing or tumorigenesis. Astrocytes are known to play major roles in wound healing in the CNS. To elucidate the roles of TNC in wound closure by astrocytes, we have examined the morphological changes of cultured astrocytes in a scratch wound assay and measured the content of soluble TNC released into the medium. We have also localized the expression of TNC mRNA, TNC, glial fibrillary acidic protein (GFAP), vimentin and integrin beta1. After wounding, glial cells rapidly released the largest TNC isoform and proliferated in the border zones. Subsequently, they became polarized with unidirectional processes and finally migrated toward the denuded area. The proliferating border zone cells and pre-migratory cells intensely expressed TNC mRNA, TNC-, vimentin-, GFAP- and integrin beta1-like immunoreactivity, while the migratory cells showed generally reduced expression except the front. Exogenous TNC enhanced cell proliferation and migration, while functional blocking with anti-TNC or anti integrin beta1 antibody reduced both of them. These results suggest that mechanical injury induces boundary astrocytes to produce and release TNC that promotes cell proliferation and migration via integrin beta1 in an autocrine/paracrine fashion. PMID- 15780468 TI - Selective cholinergic denervation, independent from oxidative stress, in a mouse model of Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by increases in amyloid-beta (Abeta) peptides, neurofibrillary tangles, oxidative stress and cholinergic deficits. However, the selectivity of these deficits and their relation with the Abeta pathology or oxidative stress remain unclear. We therefore investigated amyloidosis-related changes in acetylcholine (ACh) and serotonin (5-HT) innervations of hippocampus and parietal cortex by quantitative choline acetyltransferase (ChAT) and 5-HT immunocytochemistry, in 6, 12/14 and 18 month old transgenic mice carrying familial AD-linked mutations (hAPP(Sw,Ind)). Further, using manganese superoxide dismutase (MnSOD) and nitrotyrosine immunoreactivity as markers, we evaluated the relationship between oxidative stress and the ACh deficit in 18 month-old mice. Thioflavin-positive Abeta plaques were seen in both regions at all ages; they were more numerous in hippocampus and increased in number (>15-fold) and size as a function of age. A majority of plaques exhibited or were surrounded by increased MnSOD immunoreactivity, and dystrophic ACh or 5-HT axons were seen in their immediate vicinity. Counts of immunoreactive axon varicosities revealed significant decreases in ACh innervation, with a sparing of the 5-HT, even in aged mice. First apparent in hippocampus, the loss of ACh terminals was in the order of 20% at 12/14 months, and not significantly greater (26%) at 18 months. In parietal cortex, the ACh denervation was significant at 18 months only, averaging 24% across the different layers. Despite increased perivascular MnSOD immunoreactivity, there was no evidence of dystrophic ACh varicosities or their accentuated loss in the perivascular area. Moreover, there was virtually no sign of tyrosine nitration in ChAT nerve terminals or neuronal cell bodies. These data suggest that aggregated Abeta exerts an early, non-selective and focal neurotoxic effect on both ACh and 5-HT axons, but that a selective, plaque- and oxidative stress-independent diffuse cholinotoxicity, most likely caused by soluble Abeta assemblies, is responsible for the hippocampal and cortical ACh denervation. PMID- 15780470 TI - Functional GABA(B) receptors are expressed at the cone photoreceptor terminals in bullfrog retina. AB - GABA(B) receptors at the cone terminals in bullfrog retina were characterized by immunocytochemical and whole-cell patch clamp techniques in retinal slice preparations. Somata, axons and synaptic terminals (pedicles) of cones were both GABA(B) receptor (GABA(B)R) 1 and GABA(B)R2 immunoreactive. Physiologically, barium/calcium currents of cones to voltage steps were significantly reduced in size when GABA was puffed to cone terminals in the presence of picrotoxin that is supposed to block both GABA(A) and GABA(C) receptors. Similar reduction in barium currents was obtained with puff application of baclofen to cone terminals. These results suggest the presence of functional GABA(B) receptors at the bullfrog cone terminals. Suppression of barium currents of cones by baclofen was dose dependent. Moreover, barium currents of cones were potentiated by background illumination, as compared with those recorded in the dark. 6,7-Dinitroquinoxaline 2,3-dione, an antagonist of non-NMDA receptors that hyperpolarizes horizontal cells and reduces GABA release from these cells, and saclofen, a GABA(B) receptor antagonist, both potentiated barium currents of cones in the dark, thereby mimicking the effects of background illumination. It is suggested that changes in calcium influx into the cone synaptic terminals due to activation of GABA(B) receptors may provide a negative feedback mechanism for regulating signal transmission between cones and second-order neurons in the retina by modifying the amount of glutamate released from the cones. PMID- 15780471 TI - Neuroprotection of adult rat dorsal root ganglion neurons by combined hypothermia and alkalinization against prolonged ischemia. AB - Ischemia and ischemia-induced secondary events, such as acidosis and excessive activation of receptors by amino acids, trigger neuron death. The isolation and dissociation of dorsal root ganglion (DRG) involves time during which the neurons are ischemic due to being densely packed within the intact DRG and surrounded by a connective tissue coat. Thus, the longer the time between killing the host animal and when the DRG are dissociated, the longer the neurons are ischemic and exposed to ischemia-induced secondary causes of neuron death. It is well established that hypothermia and alkalinization each separately protect neurons from ischemia and ischemia-induced secondary causes of neuron death, but there are no data on the neuroprotection provided by simultaneous hypothermia and alkalinization. The present experiments were designed to determine the combination of hypothermic and alkaline conditions that yield the largest number of viable neurons dissociated from intact DRG maintained ischemic for up to 4 h. Hypothermia (20 degrees C>15 degrees C>37 degrees C) and alkalinization (pH 9.3>pH 8.3>pH 7.4) increased the yield of viable neurons compared with the yield from DRG maintained under physiological conditions. Hypothermia and alkalinization combined (20 degrees C/pH 9.3) provided the greatest neuroprotection with a yield of viable neurons after 1 h of ischemia 2.5-fold larger than that from DRG maintained under physiological conditions (37 degrees C/pH 7.6). Over 4 h of ischemia, the yield of viable neurons from DRG maintained under both hypothermic/alkaline and physiological conditions decreased in a linear manner, but those at 20 degrees C/pH 9.3 had a 4.5-fold greater yield of viable neurons than those at 37 degrees C/pH 7.6. Thus, combined hypothermia and alkalinization provide significantly greater protection against ischemia and ischemia-induced secondary causes of neuron death than either alone. PMID- 15780472 TI - Intraneuronal Abeta, non-amyloid aggregates and neurodegeneration in a Drosophila model of Alzheimer's disease. AB - We have developed models of Alzheimer's disease in Drosophila melanogaster by expressing the Abeta peptides that accumulate in human disease. Expression of wild-type and Arctic mutant (Glu22Gly) Abeta(1-42) peptides in Drosophila neural tissue results in intracellular Abeta accumulation followed by non-amyloid aggregates that resemble diffuse plaques. These histological changes are associated with progressive locomotor deficits and vacuolation of the brain and premature death of the flies. The severity of the neurodegeneration is proportional to the propensity of the expressed Abeta peptide to form oligomers. The fly phenotype is rescued by treatment with Congo Red that reduces Abeta aggregation in vitro. Our model demonstrates that intracellular accumulation and non-amyloid aggregates of Abeta are sufficient to cause the neurodegeneration of Alzheimer's disease. Moreover it provides a platform to dissect the pathways of neurodegeneration in Alzheimer's disease and to develop novel therapeutic interventions. PMID- 15780473 TI - The laminar histamine receptor system in human prefrontal cortex suggests multiple levels of histaminergic regulation. AB - Human prefrontal cortex is essential for high brain functions and its activity is modulated by multiple neurotransmitters, including histamine. However, the histamine receptors in this brain area have not been systematically studied so far. In situ hybridization and receptor binding autoradiography were employed to map and quantify the mRNA expression and receptor binding of three of the four histamine receptors (H(1), H(2), H(3)). mRNA expression and receptor binding of these three histamine receptors displayed characteristic laminar distribution patterns. Both H(1) and H(3) receptor mRNAs were mainly expressed in the deeper layers (H(1) in laminae V and VI; H(3) in lamina V), where most of the corticothalamic projections originate, whereas H(2) receptor mRNA was primarily expressed in the superficial layer II. Receptor ligand binding of these three histamine receptors displayed relatively even distribution patterns throughout the gray matter. However, higher densities of H(1) and H(3) receptor radioligand binding sites were seen in the middle layers III and IV that receive abundant thalamic inputs and where some of the apical dendrites of the deep-layer pyramidal neurons terminate, whereas higher density of H(2) receptor radioligand binding sites was seen in the superficial layers I-III. The results, together with data on histaminergic regulation of thalamic oscillations suggest that histamine regulates both cortico-cortical and thalamo-cortical circuits. As histamine receptors are also abundant in thalamus, histamine may be involved also in human diseases of the thalamocortical system. PMID- 15780474 TI - Serotonin-related gene expression in female monkeys with individual sensitivity to stress. AB - Female cynomolgus monkeys exhibit different degrees of reproductive dysfunction with moderate metabolic and psychosocial stress. In this study, the expression of four genes pivotal to serotonin neural function was assessed in monkeys previously categorized as highly stress resistant (n=3; normal menstrual cyclicity through two stress cycles), medium stress resistant (n=5; ovulatory in the first stress cycle but anovulatory in the second stress cycle), or low stress resistant (i.e. stress-sensitive; n=4; anovulatory as soon as stress is initiated). In situ hybridization and quantitative image analysis was used to measure mRNAs coding for SERT (serotonin transporter), 5HT1A autoreceptor, MAO-A and MAO-B (monoamine oxidases) at six levels of the dorsal raphe nucleus (DRN). Optical density (OD) and positive pixel area were measured with NIH Image software. In addition, serotonin neurons were immunostained and counted at three levels of the DRN. Finally, each animal was genotyped for the serotonin transporter long polymorphic region (5HTTLPR). Stress sensitive animals had lower expression of SERT mRNA in the caudal region of the DRN (P<0.04). SERT mRNA OD in the caudal DRN was positively correlated with serum progesterone during a pre stress control cycle (P<0.0007). 5HT1A mRNA OD signal tended to decline in the stress-sensitive group, but statistical difference between averages was lacking in analysis of variance. However, 5HT1A mRNA signal was positively correlated with control cycle progesterone (P<0.009). There was significantly less MAO-A mRNA signal in the stress-sensitive group (P<0.007) and MAO-A OD was positively correlated with progesterone from a pre-stress control cycle (P<0.007). MAO-B mRNA exhibited a similar downward trend in the stress-sensitive group. MAO-B OD also correlated with control cycle progesterone (P<0.003). There were significantly fewer serotonin neurons in the stress-sensitive group. All animals contained only the long form of the 5HTTLPR. Thus, all serotonin-related mRNAs examined in the dorsal raphe to date were lower (SERT, MAO-A) or exhibited a lower trend (5HT1A, MAO-B) in the stress sensitive animals, which probably reflects the lower number of serotonin neurons present. PMID- 15780475 TI - Differential effects of methamphetamine on expression of neuropeptide Y mRNA in hypothalamus and on serum leptin and ghrelin concentrations in ad libitum-fed and schedule-fed rats. AB - Relatively little is known concerning the interaction of psychostimulants with hypothalamic neuropeptide systems or metabolic hormones implicated in regulation of energy balance. The present studies tested whether methamphetamine alters the expression of neuropeptide Y (NPY) and agouti-related peptide (AgRP), two important orexigenic neuropeptides, or proopiomelanocortin (POMC), the precursor for the anorexigenic peptide alpha-melanocyte-stimulating hormone, or the secretion of leptin, insulin and ghrelin, concomitant with inhibition of food intake. Female rats were either fed ad libitum (AL) or placed on a scheduled feeding (SF) regimen, with access to food limited to 4 h/day. Administration of (+/-)-methamphetamine (7.5 mg/kg, i.p.) 2 h prior to food presentation significantly inhibited food intake in SF animals, but did not affect intake in AL animals. In a separate study, AL and SF animals were killed just prior to expected food presentation, and expression of NPY, AgRP and POMC mRNAs in hypothalamus was determined using in situ hybridisation; concentrations of leptin, insulin and ghrelin in serum were determined with radioimmunoassays. In saline-treated, SF controls, NPY and AgRP mRNA expression in arcuate nucleus and serum ghrelin were significantly elevated, and serum leptin and insulin were significantly reduced. Methamphetamine reversed the up-regulation of NPY mRNA expression observed in the SF condition, without affecting AgRP mRNA or the serum concentrations of metabolic hormones. However, in AL animals, NPY mRNA expression in arcuate and dorsomedial nuclei was significantly increased by methamphetamine, which also reduced serum leptin and insulin and increased serum ghrelin concentrations. These findings suggest that the inhibition of NPY expression in SF animals may be a mechanism underlying the anorexigenic effect of methamphetamine seen in this condition. The increase in NPY expression produced by methamphetamine in AL animals may be mediated by the ability of this drug to decrease secretion of leptin and insulin and increase secretion of ghrelin. PMID- 15780476 TI - Reduction of a vascular endothelial growth factor receptor, fetal liver kinase-1, by antisense oligonucleotides induces motor neuron death in rat spinal cord exposed to hypoxia. AB - Vascular endothelial growth factor (VEGF) is reported to play a neuroprotective role through a VEGF receptor, fetal liver kinase-1 (Flk-1) in vitro. We investigated whether reduction of Flk-1 could induce motor neuron loss in rat spinal cord by inhibiting the expression of Flk-1 in rat spinal cord using antisense oligodeoxynucleotides (ODNs) against the Flk-1 receptor. Rat spinal cord was repetitively exposed to 12% hypoxia, and the change of the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway and the mitogen-activated protein kinase kinase (MEK)/extracellular-signal-regulated kinase (ERK) pathway was examined. Intrathecal infusion of Flk-1 antisense ODNs for 7 days suppressed almost completely Flk-1 expression in the lumbar segment of the spinal cord and was followed by a hypoxic challenge with 12% oxygen for 1 h that was repeated for 7 more days. In the lumbar segment, we observed that reduced Flk-1 expression and hypoxic challenge for 7 days resulted in approximately 50% loss of motor neurons, in which the activation of Akt and ERK, that is, increased levels of phosphorylated-Akt and of phosphorylated-ERK by hypoxia, was markedly inhibited. In contrast, the reduction of Flk-1 expression alone did not induce motor neuron loss. These results suggest that VEGF exerts its protective effect on motor neurons against hypoxia-induced toxicity by the Flk-1 receptor through the PI3 K/Akt and the MEK/ERK signaling pathways. PMID- 15780477 TI - Induction of plasminogen activator inhibitor-1 and -2 in dorsal root ganglion neurons after peripheral nerve injury. AB - We have previously found that tissue type and urokinase type plasminogen activators (tPA and uPA) are induced in dorsal root ganglia (DRG) neurons after peripheral axotomy and that tPA plays crucial roles in generating neuropathic pain. Here we examined whether the plasminogen activator inhibitor-1 and -2 (PAI 1 and PAI-2) mRNA, endogenous inhibitors of tPA and uPA, are induced in the DRG following sciatic nerve transection. L4 and L5 DRG sections were examined using in situ hybridization histochemistry. The results showed that both PAI-1 and PAI 2 mRNA were up-regulated in DRG neurons within 1 day, and peaked at 1-3 days, after injury. Reduction of these mRNA was observed from 7 days after injury. The precise expression patterns of PAI-1 and PAI-2 mRNA at 3 days after axotomy revealed that PAI-1 mRNA was observed in predominantly small neurons, while much of the PAI-2 mRNA was expressed in large neurons. Double-labeling analysis of these mRNAs with activated transcription factor 3, known as an injury marker, revealed that most PAI-1 and PAI-2 mRNAs was induced in injured neurons. Co expression of PAI-1, 2 with tPA and uPA in DRG neurons suggests that these inhibitors may act in an autocrine manner to modulate extracellular proteolytic activity after nerve injury. PMID- 15780478 TI - Mechanical and heat sensitization of cutaneous nociceptors in rats with experimental peripheral neuropathy. AB - This study examined whether or not the properties of cutaneous nociceptive fibers are altered in the neuropathic state by comparing lumbars 5 and 6 spinal nerve ligation (SNL) rats with sham-operated controls. The rats with the unilateral SNL developed mechanical allodynia in the ipsilateral hind limb, whereas the sham group did not. Two to 5 weeks after the neuropathic or sham surgery, rats were subjected to single fiber-recording experiments to examine the properties of afferent fibers in the sural and plantar nerves. A total of 224 afferents in the C- and Adelta-ranges were characterized in the neuropathic and sham groups. Spontaneous activity was observed in 16 of 155 fibers in the neuropathic group and one of 69 fibers in the sham group. The response threshold of both the C- and Adelta-fibers to mechanical stimuli was lower in the neuropathic group than the sham group. The afferent fibers responsive to heat stimuli were all C-fibers, and none were Adelta-fibers. The response threshold of the C-fibers to the heat stimuli was lower in the neuropathic group than the sham group. The magnitude of the responses of both C- and Adelta-fibers to the suprathreshold intensity of the mechanical stimulus was greater in the neuropathic group than the sham group. However, the magnitude of the responses of C-fibers to the suprathreshold intensity of the heat stimulus in the neuropathic group was not different from that in the sham group. These results suggest that after a partial peripheral nerve injury, the nociceptors on the skin supplied by an uninjured nerve become sensitized to both mechanical and heat stimuli. This nociceptor sensitization can contribute to neuropathic pain. PMID- 15780479 TI - The inferior colliculus of the rat: a quantitative analysis of monaural frequency response areas. AB - Frequency response areas (FRAs) were measured for 237 single units in the inferior colliculus (IC) of urethane-anesthetized pigmented rats using monaural pure-tone stimulation. Based on qualitative criteria [J Neurosci 21 (2001) 7303], FRAs were classified as V-shaped in 69% of neurons, non-V-shaped in 29%, and unclassifiable in the remaining 2%. Non-V-shaped FRAs were heterogeneous, comprising a number of subtypes including narrow, closed, low- and high-tilt, multipeaked, U-shaped, mosaic and inhibitory. To complement this subjective classification, we applied quantitative measures used by others (e.g. [J Neurophysiol 84 (2000) 1012]), including the inverse slope of the upper and lower FRA borders, Q-values, and other measures of bandwidth. The results suggest that FRAs in the rat IC are best described as forming a continuous distribution among subtypes, rather than clustering into discrete categories. Moreover, there is a broad range of frequency tuning characteristics and FRA types across the entire frequency spectrum. Within this general pattern, however, there are some frequency-specific differences in FRA type distribution. The relative proportion of V-shaped FRAs was greatest at the high and low ends of the auditory range, with the highest proportion of non-V-shaped FRAs in the mid-range from 6 to 12 kHz. For most neurons with multipeaked FRAs, the peak frequencies were not harmonically related. Frequency tuning in the pigmented rat IC is generally similar to that in other species. Comparison of Q values across auditory nuclei shows little evidence that FRAs are sharpened at levels above the auditory nerve. Rather, there is a broad range of frequency tuning properties at each level. PMID- 15780480 TI - Activation of dopaminergic neurotransmission in the medial prefrontal cortex by N methyl-d-aspartate stimulation of the ventral hippocampus in rats. AB - Many behavioral functions-including sensorimotor, attentional, memory, and emotional processes-have been associated with hippocampal processes and with dopamine transmission in the medial prefrontal cortex (mPFC). This suggests a functional interaction between hippocampus and prefrontal dopamine. The anatomical substrate for such an interaction is the intimate interconnection between the ventral hippocampus and the dopamine innervation of the mPFC. The present study yielded direct neurochemical evidence for an interaction between ventral hippocampus and prefrontal dopamine transmission in rats by demonstrating that subconvulsive stimulation of the ventral hippocampus with N-methyl-d aspartate (NMDA; 0.5 mug/side) activates dopamine transmission in the mPFC. Postmortem measurements revealed that bilateral NMDA stimulation of the ventral hippocampus, resulting in locomotor hyperactivity, increased the homovanillic acid/dopamine ratio, an index of dopamine transmission, in the mPFC; indices of dopamine transmission in any of five additionally examined forebrain regions (amygdala, nucleus accumbens shell/core, lateral prefrontal cortex, caudate putamen) were unaltered. In vivo microdialysis measurements in freely moving rats corroborated the suggested activation of prefrontal dopamine transmission by demonstrating that unilateral NMDA stimulation of the ventral hippocampus increased extracellular dopamine in the ipsilateral mPFC. The suggested influence of the ventral hippocampus on prefrontal dopamine may be an important mechanism for hippocampo-prefrontal interactions in normal behavioral processes. Moreover, it indicates that aberrant hippocampal activity, as found in neuropsychiatric diseases, such as schizophrenia and mood disorders, may contribute to disruption of certain cognitive and emotional functions which are extremely sensitive to imbalanced prefrontal dopamine transmission. PMID- 15780481 TI - Excessive crying and gastro-oesophageal reflux disease in infants: misalignment of biology and culture. AB - Excessive crying is the most common problem presenting to the doctor in the first months of life in western industrialised societies, affecting up to 30% of infants. There has been an exponential increase in the diagnosis of gastro oesophageal reflux disease (GORD) in babies who cry excessively over the past few decades, and many parents believe their crying infant "has reflux". This paper proposes that culturocentric assumptions have confused interpretation of research into GORD, and re-examines the findings of GORD research from the perspective of evolutionary biology. Evolutionary biologists argue that the human infant is an exterogestate foetus for at least the first six months of life, dependent on maternal co-regulation for optimal physiological function. However, infant-care practices in western industrialised societies shifted towards an emphasis on infant autonomy at the time of the Industrial Revolution. From the perspective of evolutionary biology, a misalignment between western culture and the biological expectations of the infant developed over two million years of evolution may result in excessive crying in less adapted babies. The key biocultural factors that impact on infant distress are feeding management, parental responsiveness, sensory nourishment and sleep management. When the concept of the human infant as an exterogestate foetus is integrated with the findings of GORD research, a hypothesis and its corollary emerge. This hypothesis proposes that infant GORD is a physiological manifestation of misalignment between biology and culture, and proposes, as a corollary, that if the impact of biocultural factors upon the physiology of otherwise well crying babies is not addressed in the first months of life, populations of infants who cry excessively may be predisposed to GORD after three to four months of age. If this hypothesis is correct, an integrated clinical approach to crying babies less than three to four months of age that considers feeding management (e.g., frequent feeds, breast- or bottle-feeding technique, referral to a lactation consultant, cow's milk allergy), parental responsiveness (e.g., prompt response to infant cues), sensory nourishment (e.g., sling or backpack, walks, massage) and sleep management (e.g., nocturnal co sleeping) should, firstly, decrease crying when applied to infants less than three to four months of age, and secondly, decrease the incidence of GORD in these infants once they are older than three to four months of age. Thirdly, if this hypothesis is correct, combining the integrated approach with pharmaceutical intervention should improve outcomes in infants diagnosed with GORD. PMID- 15780482 TI - Ecstasy (MDMA) mimics the post-orgasmic state: impairment of sexual drive and function during acute MDMA-effects may be due to increased prolactin secretion. AB - Methylenedioxymethamphetamine (MDMA or "Ecstasy") is a major stimulant drug of abuse worldwide. MDMA produces euphoria, enhances interpersonal communication and feelings of closeness with others. In contrast to the induced emotions of affection and sensual enhancement, clinical studies show that it impairs sexual drive and functioning. In drug-free humans, sexual stimulation with orgasm induces a pronounced secretion of prolactin, which may mediate the post-orgasmic state. The phenomenological features of the psychological state induced by MDMA show some similarities with features of the post-orgasmic state. In addition, MDMA also induces a prominent increase of prolactin plasma levels with a similar time kinetic compared to the post-orgasmic prolactin increase. Here, we present the hypothesis that the impairment of sexual parameters after MDMA may be mediated by increased prolactin. PMID- 15780483 TI - Can worms defend our hearts? Chronic helminthic infections may attenuate the development of cardiovascular diseases. AB - The established risk factors for atherosclerosis fail to fully explain the extent and severity of coronary artery diseases in 50% of the patients. Thus, the causative agents and processes, which may be involved in the pathogenesis of atherosclerosis, are being sought. Notoriously, atherosclerosis and cardiovascular event rates are much lower in developing countries. Clinically, severe infections by intracellular pathogens are widespread mostly in developing countries with poor sanitation, nutrition and massive worm infections. A link between atherosclerosis and helminth infections has never been examined. Based on the present knowledge of immune and infectious mechanisms related to atherosclerosis, it is proposed that chronic helminthic infections can have a significant bearing on the epidemiology of cardiovascular diseases. How can helminthic infections affect the cardiovascular risk? (1) Helminths evade or suppress host immune responses, by producing anti-inflammatory and other immunomodulatory molecules. (2) Helminths induce chronic Th2 activation, which can modify cytokine profiles and immunological responses to heat shock proteins, Chlamydia pneumoniae and cytomegalovirus. (3) The chronic Th2 profile may modulate monocyte activation and chemotaxis to inflammatory sites (atherosclerotic plaques). (4) Chronic Th2 activation may lead to a cytokine profile that could be beneficial for attenuation of atherosclerosis development (upregulation of IL-4, IL-10 and IL-13 and downregulation of proinflammatory cytokines). (5) Helminthic infections may reduce plasma LDL level not only by affecting the host nutrition, but also via modulation of naturally occurring antibodies to cholesterol. Studies are needed to clarify these suggestions. If the hypothesis that helminthic infections impact atherosclerosis is correct, it should be taken into consideration in atherosclerosis immunomodulation therapy and especially in the design of vaccines and vaccine trials. PMID- 15780484 TI - Effects on the serotoninergic system in sub-acute transmissible spongiform encephalopathies: current data, hypotheses, suggestions for experimentation. AB - Sub-acute transmissible spongiform encephalopathies (TSEs), or prion diseases, are affections in which little is known of their etiology. The predominant theory stipulates that an abnormal protease-resistant prion protein (PrPres) would be infectious by directly inducing its defective conformation to the normal native protein (PrPC). The function of PrPC remains unknown. The preferred localization of PrPC at the level of the synapses supposes a function in neuronal transmission. Several neurotransmitter systems (acetylcholine, GABA, dopamine, etc.) are damaged in TSEs, mainly the serotonin (5-HT) system. At a hypothetical level, PrPC would play a trophic and functional role by regulating the capture of amino acid precursors of neurotransmitters and the functions of neuroreceptors, in particular regarding tryptophan and 5-HT receptors. By comparison with the modes of action of Ras proteins and of the envelope glycoprotein of jaagsiekte sheep retrovirus, the adaptation of an oncogenic model is suggested for the mode of action of PrPres. The sequence of events could be the following: capture of PrPres and forming of an abnormal receptor, chronic disturbance of transduction pathways, more particularly of the phosphatidylinositol-3 kinase (PI-3K)/protein kinase B (Akt)/glycogen synthetase kinase 3 (GSK 3)/Wnt-beta catenin pathway, deregulation of the PrP gene and infrequent and transitory forming of abnormal RNA messengers and, finally, the forming of abnormal proteins and the deterioration of the serotoninergic system. The involvement of endogenous nucleic acids is supposed. The infectious agent of TSEs could be an ancestral form of retrovirus, such as a retrotransposon using the prion protein as an envelope glycoprotein. Pharmacological tests, by comparison with a rare disease of unknown etiology in cattle, bovine spastic paresis, are suggested with the amino acid precursors of neuromediators (tryptophan, tyrosine, glutamic acid, etc.) and with lithium, neuroprotector and regulator of the serotonergic system. PMID- 15780485 TI - Why don't humans get scrapie from eating sheep? A possible explanation based on secondary structure predictions. AB - In an effort to find a structural explanation for the lack of direct transmission of scrapie from sheep to humans, secondary structure predictions are used to locate the segments of the prion sequence which may be involved in the transformation from the normal form of the prion protein, which has high helix content, to the pathogenic form, which has high beta-sheet content. The Chou Fasman algorithm, which calculates propensities for both helix and sheet formation, was used to predict the secondary structures of the scrapie-resistant and the scrapie-susceptible variants of the ovine prion protein. The scrapie susceptible variant, which has a glutamine at residue position 168 (human prion protein numbering), is predicted to have a propensity for sheet formation in that region of the molecule, while the scrapie-resistant variant, which has an arginine at position 168, does not. The valine at position 133, additionally present in the ovine variant which is the most susceptible to scrapie, is predicted to result in even more sheet formation. When the predicted secondary structure of the human prion protein is compared to those of the ovine prion protein variants, the human protein is found to be most similar to the scrapie resistant variant. This result is proposed to provide a possible explanation for the observation that scrapie is not directly transmitted from sheep to humans. PMID- 15780486 TI - Inflammation may be a bridge connecting hypertension and atherosclerosis. AB - Pathogenesis of the atherosclerotic process is deemed as multi-factorial, and characterized by chronic inflammatory response. Although hypertension is known to be one of the most important risk factors for atherosclerosis in causasians, its relative contribution to early atherosclerosis are still unknown. Increased evidence has indicated that hypertension, through the vasoactive peptides, such as angiotensin and endothelin-1, promotes and accelerates the atherosclerotic process via inflammatory mechanisms. In animal and human studies pro-inflammatory properties of angiotensin II has been demonstrated in large conduit and small arteries, in the kidney as well as in the heart. Activation of oxidative stress by angiotensin II is a key component of this process. Angiotensin II stimulates nicotinamide adenine dinucleotide phosphate/nicotinamide adenine dinucleotide oxidase in endothelium, smooth muscle cells, and the adventitia of blood vessel to generate reactive oxygen species, leading to endothelial dysfunction, growth, and inflammation, upregulation of endothelin-1, adhesion molecules, nuclear factor-kappa B, and other inflammatory mediators, as well as increased breakdown of nitric oxide and uncoupling of nitric oxide synthase, contribute to the progression of vascular disease and atherogenesis. In addition, recent advances concerning role of endothelin-1 as another important mediator of chronic inflammation in the vascular wall has been documented, and relationship between endothelin-1 and angiotensin II on vascular inflammation demonstrated. Inflammatory mechanisms, therefore, are important participants in the pathophysiology of hypertension-related cardiovascular disease, including atherosclerosis. In experimental models as well as human studies of atherosclerosis, angiotensin converting enzyme inhibitors or angiotensin II receptor blockers have demonstrated the ability to prevent or reverse the progression of atherosclerosis, which was in part associated with decreased expression of inflammatory mediators and improve endothelial functions. Based on those increasing evidence, we hypothesize that inflammation may be a bridge connecting hypertension and atherosclerosis. PMID- 15780487 TI - Endogenous psychoactive tryptamines reconsidered: an anxiolytic role for dimethyltryptamine. AB - The presence of the potent hallucinogenic psychoactive chemical N,N dimethyltryptamine (DMT) in the human body has puzzled scientists for decades. Endogenous DMT was investigated in the 1960s and 1970s and it was proposed that DMT was involved in psychosis and schizophrenia. This hypothesis developed from comparisons of the blood and urine of schizophrenic and control subjects. However, much of this research proved inconclusive and conventional thinking has since held that trace levels of DMT, and other endogenous psychoactive tryptamines, are insignificant metabolic byproducts. The recent discovery of a G protein-coupled, human trace amine receptor has triggered a reappraisal of the role of compounds present in limited concentrations in biological systems. Interestingly enough, DMT and other psychoactive tryptamine hallucinogens elicit a robust response at the trace amine receptor. While it is currently accepted that serotonin 5-HT(2A) receptors play a pivotal role in the activity of hallucinogenic/psychedelic compounds, we propose that the effects induced by exogenous DMT administration, especially at low doses, are due in part to activity at the trace amine receptor. Furthermore, we suggest that endogenous DMT interacts with the TA receptor to produce a calm and relaxed mental state, which may suppress, rather than promote, symptoms of psychosis. This hypothesis may help explain the inconsistency in the early analysis of endogenous DMT in humans. Finally, we propose that amphetamine action at the TA receptor may contribute to the calming effects of amphetamine and related drugs, especially at low doses. PMID- 15780489 TI - Improvement of cervical dystonia: possible role of transcranial magnetic stimulation simulating sensory tricks effect. AB - Idiopathic cervical dystonia (ICD) is the most common form of focal dystonia. A characteristic and unique diagnostic feature is the presence of "sensory tricks", for example, a finger placed on the chin to neutralize the spasm. Although approximately 70% of patients with ICD experience effective sensory tricks, the exact mechanism of these tricks is still unknown. However, recent evidence suggests that higher sensorimotor integration processes are involved. A study using H2(15)O positron emission tomography demonstrated that the application of sensory trick stimulus, resulting in a near-neutral head position, led to an increased activation mainly of the superior and inferior parietal lobules (ipsilateral to head turn) and bilateral occipital cortex and to a decreased activity of the supplementary motor area and the primary sensorimotor cortex (contralateral to head turn). Since transcranial magnetic stimulation (TMS) is an experimental device with the ability to excite or depress the neural circuits, we hypothesize that the use of TMS of specific parameters to specific brain areas (as above) may produce an effect similar to sensory tricks resulting in the relief of spasms and the improvement of cervical dystonia. PMID- 15780488 TI - Therapeutic value of antidepressants in asthma. AB - Laboratory and clinical data provide evidence that a biological linkage exists between asthma and depression. Cytokines are key molecules in both diseases. They promote allergic reaction as well as depressive symptomatology. Antidepressants may have a therapeutic role in asthma by suppressing production of proinflammatory cytokines, inducing production of anti-inflammatory ones and preventing their brain effects. Most antidepressants also induce adaptive changes in central monoaminergic neurotransmission, which itself might modulate immune reactivity and central actions of cytokines. Antidepressants may also have direct effects on the immune cells. Their impact on hypothalamus-pituitary-adrenal axis is discussed. Antidepressants are expected to terminate the cascade of inflammatory events in other inflammatory diseases as well. The use of antidepressants in experimental clinical trials in patients with asthma is suggested. PMID- 15780490 TI - The potential importance of steroids in the treatment of autistic spectrum disorders and other disorders involving mercury toxicity. AB - Autism is a neurodevelopmental disorder that according to the Centers for Disease Control and Prevention (CDC) affects 1 in 150 children in the United States. Autism is characterized by impairments in social relatedness and communication, repetitive behaviors, abnormal movements, and sensory dysfunction. Recently emerging evidence suggests that mercury, especially from childhood vaccines, appears to be a factor in the development of the autistic disorders, and that autistic children have higher than normal body-burdens of mercury. In considering mercury toxicity, it has previously been shown that testosterone significantly potentates mercury toxicity, whereas estrogen is protective. Examination of autistic children has shown that the severity of autistic disorders correlates with the amount of testosterone present in the amniotic fluid, and an examination of a case-series of autistic children has shown that some have plasma testosterone levels that were significantly elevated in comparison neurotypical control children. A review of some of the current biomedical therapies for autistics, such as glutathione and cysteine, chelation, secretin, and growth hormone, suggests that they may in fact lower testosterone levels. We put forward the medical hypothesis that autistic disorders, in fact, represents a form of testosterone mercury toxicity, and based upon this observation, one can design novel treatments for autistics directed towards higher testosterone levels in autistic children. We suggest a series of experiments that need to be conducted in order to evaluate the exact mechanisms for mercury-testosterone toxicity, and various types of clinical manipulations that may be employed to control testosterone levels. It is hoped by devising therapies that address the steroid hormone pathways, in addition to the current treatments that successful lower heavy metal body-burdens of mercury, will work synergistically to improve clinical outcomes. In light of the fact that there are a number of other diseases that may have a chronic mercury toxicity component, such as Alzheimer's disease, heart disease, obesity, ALS, asthma, and other various forms of autoimmune disorders, it is imperative that further research should be conducted to understand mercury-testosterone toxicity. PMID- 15780491 TI - Protein aggregation as a possible cause for pathology in a subset of familial Unverricht-Lundborg disease. AB - Loss of function mutations in the gene (CSTB) encoding human cystatin B, a widely expressed cysteine protease inhibitor, are responsible for a severe neurological disorder known as an Unverricht-Lundborg disease (EPM1). EPM1 had been linked to chromosome 21q22.3 in Finnish families and it is an autosomal recessive inherited disorder with a homozygous minisatellite expansion in the cystatin B gene (stefin B gene). This disease is difficult to treat because it is refractory to most antiepileptic drugs and using multiple medications had been unsuccessful so far. To come a step closer to understanding of the nature of this disease, especially about the events on the molecular level, in vitro properties of missense EPM1 mutant G4R were determined. It was observed that the mutant has a prolonged lag phase of fibrillation at the same protein stability, which could indicate it were more toxic to the cells. Similar experiments with the N-terminal fragment of 67 aminoacid residues are ongoing, showing higher propensity to aggregate. Therefore, a hypothesis is launched that at least in a subset of Unverricht Lundborg disease patients, cystatin B protein may aggregate in the cell. Protein aggregation can be secondary to external insults or aging, however, inherited forms of neurodegenerative diseases, such as familial Parkinson's, Huntington's or familial Alzheimer's disease, are directly linked to the mutant proteins aggregation. Protein aggregates in the form of amyloid plaques, neurofibrilary tangles, intra-cytoplasmic or intra-nuclear inclusions lead to increased production of the reactive oxygen species and dysfunction of the ubiquitine/proteasome system. Finally, mitochondrial dysfunction and cell death are observed. Certainly, it remains to be checked by experiments whether overexpression in cell culture of the missense mutants G4R and N-terminal fragment to residue 68 lead to cellular inclusions and the accompanying changes characteristic for the conformational disorders. PMID- 15780492 TI - Alzheimer disease is substantially preventable in the United States -- review of risk factors, therapy, and the prospects for an expert software system. AB - Epidemiology studies, including both regional incidence and the analysis of specific risk factors for Alzheimer's disease indicate that substantial prevention of the disease, in the 50-70 percent range, is a practical possibility for the United States. Epidemiology has identified a rich diversity of specific prevention strategies relating to nutrition, dietary supplements, lifestyle, food and environmental toxins, and in some cases medication, many of which have a capacity to reduce Alzheimer's risk by 50 percent or more. The interaction of these risk factors with brain biology is increasingly understood. In contrast, therapeutic strategies for un-prevented Alzheimer's generally prove incapable of delaying disease progression by more than 3-11 months, because extensive brain cell death occurs even in preclinical or mild cases. A public health program aimed at prevention can be fashioned with expert software packages, based on already identified risk factors. Such statistical analysis should allow the prediction of individual and group Alzheimer's risks of sufficient power to instruct the formulation of lifestyle, nutritional and environmental programs to substantially reduce disease incidence. A less satisfactory but complementary alternative is very early disease detection with therapeutic strategies focused on retardation of brain cell death, so that the person dies of another cause before the disease is clinically manifested. PMID- 15780493 TI - Neurodegeneration caused by the translation of nonsense: does macromolecular misfolding impair the synchrony of gene expression? AB - Here, it is proposed that the principal event underlying neurodegeneration occurs when cytotoxic, truncated proteins are expressed from normally-untranslated nonsense RNA and pseudogene transcripts. The proximal event occurs when a small fraction of the total pool of gene expression machinery within the cell is disrupted by rare events of macromolecular misfolding during gene expression. Macromolecular misfolding, such as beta-sheet formation of protein leading to intracellular aggregation, has been implicated in a number of neurodegerative diseases. As gene expression is a synchronised series of processes from the nucleus to the cytoplasm, should macromolecular misfolding occur in any given component of the gene expression apparatus, co-dependent gene expression processes could become disrupted. For example, should proteins misfold during their own translation, aggregates could accumulate within the translation machinery and disrupt the regulation of upstream gene expression events, such as RNA splicing or Nonsense Mediated Decay. Although only a limited amount of gene expression machinery would be affected by macromolecular misfolding, the resultant loss in fidelity could allow sufficient levels of expression of aberrant proteins from nonsense RNA and pseudogene transcripts to produce cytotoxic effects within the cell over time and ultimately lead to neurodegeneration. PMID- 15780494 TI - Antioxidants may increase the probability of developing allergic diseases and asthma. AB - In addition to genetic predisposition, a lack of triggers for Th1 immune response like exposure to infections, endotoxins and dirt in childhood are supposed to be responsible for the higher incidence of allergic rhinitis and asthma (hygiene hypothesis). In vitro, beverages rich in antioxidants like green tea and wine were found to suppress formation of Th1-type cytokine interferon-gamma. Due to the existing cross-regulatory interplay between Th1- and Th2-type immune response, these beverages may thus slow-down Th1-type immune response and thereby favour an over-production of Th2-type cytokines. Also food rich in antioxidants may increase the risk of atopic disease. Thus, not only a lack of triggers for Th1 type immune response, but also a nutrition rich in antioxidants suppressing interferon-gamma would result in a persistence of Th2-type immune response and increase the susceptibility for allergic reactions and asthma. In addition to improved hygienic standards in the past decades, also social changes including the availability of functional food and food enriched in antioxidants may have increased the prevalence of atopic diseases in Western countries. PMID- 15780495 TI - Potentiation of immunological tolerance induction in adult mice by co administration of pooled normal IgG and oral tolerogens: a potential therapeutic approach for autoimmune diseases. AB - Oral tolerance can be defined as the inability of an adult animal to produce specific antibodies or cellular immune responses upon conventional immunization, after oral antigenic administration. Recently, the oral administration of antigens has gained renewed interest because of the possibility of inducing tolerance in nonimmunized adult animals and, consequently, opening up the theoretical possibility of preventing or treating diseases caused by malfunction of the immune system. This strategy has been proven to be useful in the prevention of allergic and autoimmune diseases in rodents, as well as in the amelioration of certain autoimmune diseases in humans. Although there is experimental and clinical evidence for the usefulness of oral tolerance in medical practice, the mechanisms responsible for this phenomenon are still poorly understood, and the results obtained are not always satisfactory. Herein, we show that the thymus is required for the induction and maintenance of oral tolerance, providing evidence that it is not a pure form of clonal deletion-based peripheral tolerance. Oral tolerance could therefore depend on the formation and release to the periphery of regulatory T cells, such as gammadelta or alphabeta T cells, by the thymus. This finding may have profound implications for the treatment of autoimmune diseases, since most of them are associated with thymic hypofunction. On the other hand, due to so far unknown mechanisms, the intraperitoneal co administration of normal IgG to mice orally treated with tolerogen leads to a sustained and intense immunological tolerance, both in euthymic and thymectomized mice, including those of the lupus erythematosus-prone NZB x NZW lineage. This approach for inducing and maintaining tolerance in thymus-deficient conditions is discussed and put forth herein as a new evidence-based proposition for the therapy of autoimmune diseases. PMID- 15780496 TI - Does autoantigen administration inhibit recurrence of type 1 diabetes in transplanted islets? AB - Type 1 diabetes is a chronic disease with a subclinical prodromal period resulting from autoimmune destruction of pancreatic beta-cells. At the time of clinical symptoms of diabetes, the majority of islets have irreversibly been destroyed. Thus, the only cure for type 1 diabetes is pancreas (or islet) transplantation. To reach this goal, both allograft rejection and recurrent autoimmunity must be overcome. These have partly been achieved at the cost of lifelong immunosuppression, however, the risk to benefit ratio for immunosuppressive drugs to insulin usage remains obscure. To eliminate the need for immunosuppression, several tolerance induction protocols have been developed which particularly target alloimmune responses, whereas no tolerance induction protocol that particularly prevents the recurrence of autoimmunity has been suggested. It is hypothesized that autoantigen-based interventions may inhibit the recurrence of type 1 diabetes in transplanted pancreas (or islets) through the induction of specific tolerance to beta-cell autoantigens. PMID- 15780497 TI - Estradiol induced inhibition of 11beta-hydroxysteroid dehydrogenase 1: an explanation for the postmenopausal hormone replacement therapy effects. AB - The adverse and beneficial effects of postmenopausal hormone replacement therapy include: ischemic heart disease, stroke, pulmonary embolism, breast cancer, an increased rate of onset of asthma as well as reductions in the incidence of diabetes in women with known coronary artery disease and osteoporotic fractures. These varied effects can be explained by the down regulation of 11beta hydroxysteroid dehydrogenase by estradiol, which results in a reduction of tissue specific cortisol production. The reduction in local cortisol production which diminishes the endogenous anti-inflammatory effects, also allows for the progression of both vascular and pulmonary inflammation. The decrease in cortisol activation reduces insulin resistance and anti-proliferative effects thereby reducing the potential for diabetes but allowing for the emergence of malignancy. Furthermore, the decreased local tissue availability of cortisol reduces the tendency for the development of osteoporosis. New techniques and drugs are being developed to evaluate the modulation of 11beta-HSD1 activity. Further study should result in new ways to control both inflammation and metabolism. PMID- 15780498 TI - Incorporating protein transduction domains (PTD) within intracellular proteins associated with the 'stemness' phenotype. Novel use of such recombinant 'fusion' proteins to overcome current limitations of applying autologous adult stem cells in regenerative medicine? AB - Adult stem cells originating from post-natal tissues hold tremendous promise in regenerative medicine. Nevertheless, there are several deficiencies of adult stem cells that would limit their application in transplantation therapy, in particular their relative scarcity, restricted multi-potency and limited proliferative capacity in vitro. A possible approach to overcome these limitations would be to genetically modulate adult stem cells to strongly express genes that are closely associated with the 'stemness' phenotype. Overwhelming safety concerns would preclude the direct application of recombinant DNA technology in genetic modulation. Moreover, constitutive expression of 'stemness' genes would prevent adult stem cells from participating in tissue/organ regeneration upon transplantation. A novel alternative would be to incorporate protein transduction domains within intracellular proteins (i.e. transcription factors) that are associated with the 'stemness' phenotype. Such recombinant fusion proteins would then have the ability to translocate across the cell membrane and be internalized within the cytosol, thereby enabling them to exert a gene-modulatory effect on the cell, without any permanent genetic alteration. This would be particularly useful for maintaining the 'stemness' of adult stem cell populations during extensive ex vivo proliferation, to generate adequate cell numbers for transplantation therapy. PMID- 15780499 TI - Targeting fatty acid synthase-driven lipid rafts: a novel strategy to overcome trastuzumab resistance in breast cancer cells. AB - Trastuzumab (Herceptin) is a humanized antibody directed against the extracellular domain of the tyrosine kinase orphan receptor Her-2/neu (erbB-2) that has shown therapeutic efficacy against Her-2/neu-overexpressing breast tumors. However, less than 35% of patients with Her-2/neu-overexpressing metastatic breast cancer respond to trastuzumab as a single agent, whereas the remaining cases do not demonstrate tumor regression. Furthermore, the majority of patients who achieve an initial response generally acquire resistance within one year. Therefore, the identification of the potential mechanisms of resistance to trastuzumab can be very helpful for the development of new compounds, which might overcome that resistance and/or have additive/synergistic antitumor effect when given in association with trastuzumab. Recent studies in breast cancer cells have revealed a bi-directional connection between Her-2/neu and fatty acid synthase (FAS), a major lipogenic enzyme catalyzing the synthesis of long-chain saturated fatty acids from the 2-carbon donors malonyl-CoA and acetyl-CoA. Her-2/neu overexpression stimulates the FAS promoter and ultimately mediates increased endogenous fatty synthesis, and this Her-2/neu-mediated induction of breast cancer-associated FAS is inhibitable by trastuzumab. On the other hand, chemical FAS inhibitors as well as RNA interference-mediated silencing of FAS gene repress Her-2/neu gene expression at the transcriptional level. Moreover, specific FAS blockade synergistically sensitizes breast cancer cells carrying Her-2/neu oncogene amplification and/or overexpression to trastuzumab-induced cell growth inhibition and apoptotic cell death. Strikingly, FAS inhibition synergistically interacts with trastuzumab in Her-2/neu-negative breast cancer cells engineered to overexpress Her-2/neu, thus suggesting that the molecular linkage between FAS activity and functioning of Her-2/neu cannot be explained only on the basis of a transcriptional repression of Her-2/neu gene promoter. Interestingly, while in liver and adipose tissue FAS produces fat from excess carbon consumed as carbohydrates, which is ultimately stored as triglycerides, in epithelial cancer cells, FAS activity is mainly involved in the production of phospholipids partitioning into detergent-resistant membrane microdomains (lipid raft aggregates), which point to an active role of FAS in the deregulation of membrane functioning in tumor cells. Importantly, clusters of Her-2/neu and EGFR (erbB-1) co-localize with lipid rafts and the lipid environment in the cell membrane of breast cancer cells profoundly influences their association properties and biological functions. We hypothesize that pharmacological or small interference RNA-induced inhibition of breast cancer-associated FAS will result in major changes in the synthesis of phospholipids which, in turn, should impair a correct cellular localization of Her-2/neu at the cellular membrane of breast cancer cells. In this working model, FAS inhibition could induce a shift in the equilibrium between transport of Her-2/neu to and from the membrane favoring an increased Her-2/neu internalization followed by intracellular degradation, thus enhancing the mechanism of action of the anti-Her-2/neu antibody trastuzumab. Moreover, the inhibition of FAS-driven lipid rafts will also negatively affect EGFR-Her-2/neu cross-talk, an important mechanism of trastuzumab resistance. In summary, the specific blockade of a novel molecular linkage between FAS-regulated membrane composition and functioning of transmembrane growth factor receptors EGFR and Her-2/neu may represent a previously unrecognized therapeutic approach circumventing trastuzumab resistance in breast carcinomas. PMID- 15780500 TI - Cancer by negative heterosis: breast and ovarian cancer excess in hybrids of inbred ethnic groups. AB - Breast and ovarian cancer rates in Pakistan are significantly higher than in neighboring countries. The cancer rate discrepancies cannot be explained with discrepancies of their risk factors. We propose that observed cancer excess in Pakistan is due to cancer development by negative heterosis. Heterosis occurs when a hybrid has a phenotypic characteristic significantly different from that in either parent (hybrid vigor). At a molecular level, heterosis occurs in a heterozygote when one of the two alleles is inactivated. Gene inactivation occurs by methylation of cytosine in a promoter region of a gene. Initiation of allele inactivation is linked to the factors like stress, gender, diet, or another gene. In heterozygote, inactivation of one of the two tumor-suppressor alleles leads to monoallelic expression. This increases cancer risk in the same way the risk is increased in individual who inherit a single mutated tumor-suppressor gene (hereditary cancer syndrome). In both, cancer by heterosis and inherited cancer syndrome, cancer develops after inactivation of a second allele (second hit hypothesis). In a population, conditions that favor development of cancer by heterosis are those that favor mating of a large number of different homozygotes because they produce a large number of different heterozygotes. Among a large number of heterozygotes, there is an increased chance that some of hybrids will develop cancer by heterosis. In Pakistan, conditions were favorable for cancer development by heterosis because country has a high number of different ethnic groups and brotherhoods all of which have a higher rate of homozygosity due to a high frequency of consanguineous marriages, and marriages between members of different groups occurred because of intense population mixing. Result was birth of a large number of inter-ethnic/brotherhood hybrids (heterozygotes), some of which have developed cancer by heterosis. PMID- 15780501 TI - Homocysteinemia is a risk factor for aortic dissection. AB - There are significant associations between moderate increases in serum homocysteine and three cardiovascular diseases: ischemic heart disease, deep vein thrombosis and pulmonary embolism, and stroke. An association between the presence of abdominal aortic aneurysm and elevated homocysteine plasma levels has been indicated. Although chronic systemic hypertension is the most common factor predisposing the aorta to dissection, homocysteinemia has never been known as the risk for aortic dissection except for that with Marfan syndrome. Homocysteinemia is suggested to be the risk for aortic dissection in Marfan syndrome and spontaneous cervical artery dissection. Reduced fibrillin-1 deposition into the extracellular matrix is found not only in Marfan syndrome but also in isolated ascending aortic aneurysm and dissection. The reduced matrix deposition produces a mild form of weakness of elastic tissue, which predisposes to ascending aortic aneurysm and dissection in patients who do not have the Marfan syndrome. The defect in fibrillin-1 leads to: (1) formation of elastin that is abnormally aggregated and more easily degraded by matrix metalloproteinases than is normal elastin; (2) upregulation of the synthesis of matrix metalloproteinases; (3) progressive destruction of connective tissue by these enzymes; (4) development of thoracic aortic aneurysms. Homocysteine causes premature breakdown in the arterial elastic fibers by activation of the elastolytic activities. Irreversible homocysteinylation of long-lived proteins should lead to cumulative damage and progressive clinical manifestations, and fibrillin-1 is seen as the paradigm of extracellular connective tissue proteins that are specially susceptible to homocysteine (and presumably homocysteine thiolactone) attack. The authors hereupon propose a novel hypothesis that homocysteine plays an important role in development of aortic dissection and that homocysteinemia is one of the risk factors for aortic dissection. PMID- 15780502 TI - The role of viruses in cell fusion and its importance to evolution, invasion and metastasis of cancer clones. AB - The hypothesis described here is a logical extension of two areas of observation: First, it has been discovered that viruses (and perhaps other intracellular parasites) catalyze cell fusion as a means of cell-to-cell transmission. Effective cell-to-cell transmission appears to require: (i) induced expression of adhesion molecules on the cell surface; (ii) suppression of p53-dependent apoptosis; (iii) arrest of the cell cycle that would otherwise lead to cell death by "mitotic catastrophe". Suppression of apoptosis and cell death through "mitotic catastrophe" are important for formation of stable syncytia. Expression of Bcl-2 or a viral analogue of Bcl-2 (vBcl-2) is particularly useful to viruses because Bcl-2 both suppresses (p53-dependent) apoptosis and arrests the cell cycle through p27. Bcl-2 may also block any p53-independent cell death (e.g., mitotic catastrophe) that is initiated at the mitochondria. Second, it has been found that cell fusion plays a role in cancer clone evolution, invasion of normal cells in tissue adjacent to tumors and metastasis to remote normal tissues. Thus, it can be hypothesized that infection of cancer cells with viruses that spread by cell-to-cell transmission may coincidentally contribute to development of aggressive aneuploid clones and facilitate both invasion and metastasis of tumors. Regardless of the role of viruses, suppression of Bcl-2 may be an approach to preventing successful formation of syncytia and limiting the invasion and metastasis of tumors, thus, making surgical removal and radiation treatment more feasible. PMID- 15780503 TI - Nutritional modulation of parathyroid hormone secretion may influence risk for left ventricular hypertrophy. AB - Recent studies demonstrate low serum levels of 25-hydroxyvitamin D in patients with congestive heart failure (CHF). Although this may in part reflect reduced capacity for outdoor exercise, the possibility that poor vitamin D status increases risk for left ventricular hypertrophy (LVH), and its common sequel CHF, merits consideration. In cardiomyocytes, hormones which activate protein kinase C (PKC) -- including norepinephrine, angiotensin II, and endostatin, implicated in the pathogenesis of LVH -- induce a hypertrophic response analogous to that seen in LVH. Transgenic mice overexpressing PKC-beta2 or its upstream activator Galphaq in cardiac myofibers develop a syndrome similar to LVH. Parathyroid hormone (PTH) also activates Galphaq and PKC in cardiomyocytes, and provokes the expected hypertrophic response. Both primary and secondary hyperparathyroidism are associated with high risk for LVH. Moreover, in uncomplicated essential hypertension, left ventricular mass index has been shown to correlate very tightly with serum PTH levels, independent of blood pressure. This latter finding suggests that variations of PTH within the normal range can influence induction of LVH in at-risk subjects. If so, nutritional and lifestyle measures which modulate PTH secretion may have an impact on LVH risk. PTH secretion should be down-regulated by good vitamin D status -- achieved through supplementation or regular uv exposure -- and by vegan diets moderately low in bioavailable phosphate. Although high calcium intakes can likewise suppress PTH, they also boost renin secretion, which could have a countervailing effect on risk for LVH. Whether these nutritional measures do indeed influence LVH risk could be examined in prospective studies targeting patients at high risk, such as hypertensives. PMID- 15780504 TI - Secondary hyperparathyroidism promotes the acute phase response -- a rationale for supplemental vitamin D in prevention of vascular events in the elderly. AB - Parathyroid hormone (PTH) promotes IL-6 secretion by osteoblasts, and may also up regulate IL-6 production in the liver and adipose tissue; this may explain why serum IL-6 is markedly elevated in primary hyperparathyroidism, and low in hypoparathyroidism. IL-6 is the chief stimulus to hepatic production of many acute phase reactants, notably fibrinogen and C-reactive protein (CRP). Mild secondary hyperparathyroidism is common in elderly people, particularly at high latitudes during the winter, owing to poor vitamin D status. This may rationalize evidence that acute phase proteins show seasonal variations and are typically elevated in the elderly, whereas leisure physical activity is associated with a reduction in these proteins. In a recent clinical trial targeting elderly chronically ill patients, administration of vitamin D reduced serum levels of both CRP and IL-6; further such studies should assess the impact of physiologically meaningful doses of vitamin D on acute phase reactants in elderly subjects likely to have poor vitamin D status. Since elevations of CRP and fibrinogen may increase risk for thromboembolic vascular events, these considerations may help to explain the excess of coronary mortality observed during winter months, and suggest a role for supplemental vitamin D in preservation of vascular health. Moderate alcohol intake is associated with reduced serum PTH as well as decreased levels of CRP and fibrinogen; conceivably, modulation of PTH mediates, at least in part, the favorable impact of moderate drinking on the acute phase reactants. PMID- 15780505 TI - Marinobufagenin may mediate the adverse impact of salty diets on renal calcium retention by impairing the efficiency of renal tubular sodium-calcium exchange. AB - For reasons yet to be clarified, salt loading and plasma volume expansion decrease renal calcium retention; consequently, high-salt diets are thought to increase risk for osteoporosis and renal stones. These measures also can evoke increased adrenal production of the natriuretic factor marinobufagenin (MBG), recently implicated in the genesis of essential hypertension. MBG achieves natriuresis via potent selective inhibition of the alpha-1-type sodium pump, expressed throughout the nephron. In as much as renal calcium retention is largely dependent on efficient activity of calcium-sodium exchangers situated in the basolateral membranes of tubular epithelium, it is evident that an increased intracellular sodium concentration consequent to sodium pump inhibition could blunt the activity of these exchangers. Thus, it is postulated that MBG mediates the impact of salt loading on renal calcium retention. PMID- 15780506 TI - Supplemental arginine and high-dose folate may promote bone health by supporting the activity of endothelial-type nitric oxide synthase in bone. AB - The endothelial isoform of NO synthase promotes maintenance of bone density by stimulating osteoblastic activity while inhibiting bone catabolism; it appears to be a key mediator of the anabolic effects of mechanical loading, estrogens, and statin therapy on bone. This enzyme is susceptible to competitive inhibition by elevated systemic levels of asymmetric dimethylarginine (ADMA) encountered in vascular disorders associated with endotheliopathy; it may not be coincidental that reduced bone density has been observed in subjects afflicted with many of these disorders. Supplemental arginine has the potential to offset this adverse effect of ADMA. Superoxide production by osteoclasts may also impair bone NO synthase activity by oxidizing its cofactor tetrahydrobiopterin; high-dose folate has been shown to compensate for endothelial deficiency of this cofactor by effectively "pinch hitting" for it. These considerations suggest that supplementation with arginine as well as high-dose folate might aid maintenance of bone density by helping to preserve optimal NO synthase activity in bone cells. PMID- 15780507 TI - Evidence-based medicine is rooted in Protestant exegesis. AB - Evidence-based medicine (EbM) has been practised for about a decade now. Until now, it has generally been accepted that EbM has its roots in medical thinking of mid-19th century France. Due to the startling fact that France never was a centre of EbM, historical tradition was reconsidered. Since EbM has mainly been flourishing in Protestant countries, a qualitative historical investigation was conducted according to the approach of Max Weber's "The Protestant Ethics". Thus, it could be shown that there are three major prerequisites for EbM to evolve apart from current technical developments, such as the computer and the internet: (1) historical critical exegesis functioned as a methodology to balance contradictory passages; (2) both an equality based relationship among physicians and a Protestant concept that lay people are considered equal in the theologic debate were fundamental to EbM as a new approach of medical thinking; (3) mostly nationally funded health care systems are prone to practise EbM as they are obliged to provide health care which is both fair in access and allocation to the whole population. Against the background of historical exegesis, it has to be taken into account that EbM implies a twist in medicine towards a concept of textual criticism rather than the mere introduction of statistics. Moreover, it both relies upon and enhances a more equal relationship between physicians. PMID- 15780508 TI - Relativistic parameters of senescence. AB - The laws of biochemistry and biology are governed by parameters whose description in mathematical formulas is based on the three-dimensional space. It is a fact, however, that the life span of a cell and its specific functions, though limited, can be extended or diminished depending on the genetic code but also, on the natural pressure of the environment. The plasticity exhibited by a cellular system has been attributed to the change of the three-dimensional structure of the cell, with time being a simple measure of this change. The model of biological relativity proposed here, considers time as a flexible fourth dimension that corresponds directly to the inertial status of the cells. Two types of clocks are defined: the relativistic biological clock (RBC) and the mechanical clock (MC). In contrast to the MCs that show the astrological reference time, the time shown by the RBCs delay because it depends on cellular activity. The maximum and the expected life span of the cells and/or the organisms can be therefore relied on time transformation. One of the most important factors that can affect time flow is the energy that is produced during metabolic work. Based on this observation, RBCs can be constructed following series of theoretical experiments in order to assess biological time and life span changes. PMID- 15780509 TI - Misattributed paternity. A bias in the family studies in schizophrenia? AB - One of the few consistent findings originating from the research in schizophrenia is the high familiality in the transmission of the disease. Data from family studies have been used in the attempt to disentangle the genetic from the environmental components of this familiality. The analysis of this body of research has led several groups to the conclusion that there is a higher heritability of schizophrenia through the female rather than the male lineage and that the concordance for the illness is higher among DZ twins than siblings. These assumptions have generated a flurry of hypotheses on the aetiology of schizophrenia. This article demonstrates that the above findings can instead be explained as the result of an artefact originating from the uncertainty in the paternity of the probands. None of the studies has in fact considered that, in some populations, up to 30% of children are not genetically related to their putative fathers. This issue is potentially important for the genetic studies in all pathologies, but has a particular relevance for schizophrenia, due to the high profile acquired by family studies in the research for the aetiology in this illness. The conclusions reached in most family studies in schizophrenia should therefore be reappraised. PMID- 15780510 TI - Paradoxical strategy for treating chronic diseases where the therapeutic effect is derived from compensatory response rather than drug effect. AB - Reversing chronic conditions remains an elusive goal of medicine. The modern medical paradigm based on blocking overactive pathways or augmenting deficient pathways offers symptomatic benefit, but tolerance to therapy can develop and treatment cessation can produce rebound symptoms due to compensatory mechanisms. We propose a paradoxical strategy for treating chronic conditions based on harnessing compensatory mechanisms for therapeutic benefit. Many current drugs may be repurposed for a paradoxical indication where the therapeutic effect is derived from compensatory response, rather than drug effect. For example, although exercise is associated with acute adrenergia, paradoxical downregulation of baseline sympathovagal ratio occurs as a remodeling response. For conditions that manifest chronic sympathetic bias such as cardiovascular diseases, judicious administration of adrenergic agonists may induce compensatory downregulation of baseline sympathovagal ratio. The concept may generalize to many other diseases, especially those involving pathways which exhibit strong homeostatic tendencies such as the neurologic, immune, and endocrine systems. Careful consideration of chronobiologic features is necessary to optimize dosing strategies for modulating compensatory responses, and eccentric dosing schedules, shorter-acting formulations, or pulsatile delivery may be desirable in some cases. To what extent the effect of desensitization to current therapy is mistaken for disease progression in conditions such as diabetes, myopia, depression, and hypertension warrants investigation. The merits of combining behavioral and drug therapies such as diet-insulin therapy for diabetes and exercise-beta-blockade for cardiovascular disease should be revisited since there is a risk for exacerbating the underlying dysfunction. The reduced dynamic range of various environmental experiences and the tendency to revert to the mean through medical intervention, thermoregulation, and other modern lifestyle changes may play under-recognized roles in human diseases. Perhaps alternating agonists and antagonist may exercise the entire dynamic range of pathways and improve health. PMID- 15780511 TI - Oxytocin and osteoporosis: is there any relationship? PMID- 15780512 TI - Non-chaotic behavior of human cancers. PMID- 15780513 TI - Nicotinamide trials in diabetes intervention. Does a metabolite provide benefit? PMID- 15780514 TI - Thimerosal, micromercurialism and chronic fatigue syndrome. PMID- 15780515 TI - Estimate of the average electric field produced by a mobile telephone inside human skull. PMID- 15780516 TI - Atherosclerosis -- an infectious disease? PMID- 15780517 TI - Diabetes mellitus, acquired immune deficiency syndrome and tuberculosis. PMID- 15780518 TI - Innate properties of H-Wave device, a small fiber stimulator provides the basis for a paradigm shift of electro-therapeutic treatment of pain with increased functional restoration associated with human neuropathies by affecting tissue circulation: a hypothesis. PMID- 15780519 TI - Molecular mechanism of chemotherapeutic drug(s)-, induced autoimmunity: a mitochondrial-lysosomal axis theory. PMID- 15780520 TI - Estrogen receptor (ER) in breast cancer tissue of premenopausal patients: are some ER- findings false due to down-regulation? PMID- 15780521 TI - Could repetitive transcranial magnetic stimulation be effective in autism? PMID- 15780522 TI - Value of C-reactive protein levels and IL-6 in predicting events levels in women at increased cardiovascular risk. AB - BACKGROUND: Elevated C-reactive protein (CRP) levels due to of heightened vascular inflammatory state in vascular conditions are often associated with elevated interleukin-6 (IL-6) levels since during inflammation CRP production in the liver is induced by IL-6. It has been suggested that CRP may be a predictors of unfavourable outcome in postmenopausal women (PMW) receiving hormone replacement therapy. Because of the possible metabolic effect of hormone replacement therapy (HRT) on CRP, the relative predictive importance of CRP and IL-6 levels in PMW receiving HRT remains to be elucidated. METHODS: We measured plasma levels of CRP and IL-6 levels in 346 consecutive PMW (mean age 66+/-9 years) with cardiovascular risk >20 in 10 years followed during a 36 month period. Women underwent measurement of inflammatory cytokines at baseline and were allocated to two groups according to the willingness to take hormone replacement therapy. All women underwent a further measurement of CRP and IL-6 at 3 and 6 months. Health status was assessed by out patient visits and hospital charts. RESULTS: During 1 year follow up, three patient died, four had a major cardiovascular event, three had a unstable angina, two had a transient ischemic attack and two patients underwent PTCA. PMW with events had higher CRP levels compared with patients with no events (1.9+/-0.61 versus 1.43+/-0.21, P<0.05) but still within the limits of normal. Also baseline IL-6 plasma levels were significantly higher in PMW with events than in those without events (0.87+/-0.23 versus 0.54+/-0.18, P <0.05). The increase in CRP and IL-6 with HRT was significantly higher in patients with events than in those with no events (CRP: 81+/-12 % versus 76+/-21%, P<0.05; IL-6 9+/-3 % versus -14+/-7%, P<0.05). In a stepwise multivariate analysis, IL-6 levels resulted a stronger predictor of outcome than CRP. CRP levels were predictors of future events only after removal of IL-6 levels and presence of cardiovascular symptoms from the analysis. CRP levels were associated with an unfavourable outcome only when IL-6 levels were also elevated. The increase in CRP with HRT during follow up was not associated to an increased event rate. CONCLUSION: Our study showed that CRP levels are increased in PMW receiving HRT. Elevated IL-6 levels may identify those PMW at increased 1 year risk. CRP levels predict events only when they are coupled with elevated IL-6 levels. PMID- 15780523 TI - Differential estrogen signaling in endothelial cells upon pulsed or continuous administration. AB - While experimental evidence demonstrates that estrogen protects vascular cells, clinical trials on hormone replacement therapies (HRT) fail to report cardiovascular benefits. This discrepancy may indicate that estrogen signaling during HRT may not be fully effective in vascular cells, possibly due to the way of delivering estrogens to vascular tissues. We therefore, tested whether a different kinetics of exposure of endothelial cells to estrogens may alter the balance between transcriptional and non-transcriptional signaling. METHODS AND RESULTS: Cultured human umbilical vein endothelial cells (HUVEC) were exposed to equal amounts of 17beta-estradiol administered transiently (7 nM for 1 h) or continuously (0.29 nM for 24 h), using endothelial nitric oxide synthase (eNOS) expression and activity as functional endpoints. Transient and continuous administrations equally increased eNOS expression, implying that a short contact between hormone and cells is sufficient to trigger genomic pathways. In agreement, in ovariectomized rats pulse-like changes in plasma estradiol or stable levels equally increased aortic eNOS. More importantly, we extensively show that transient estradiol administration results in preferential recruitment of non-genomic pathways, leading to more vivid activation of MAPK and phosphatidylinositol-3 kinase (PI3K) cascades. CONCLUSIONS: Our data suggest that the kinetics of contact of estrogen with the cells could represent a sensor for estrogen receptor (ER) signaling towards non-genomic or genomic pathways. These findings deepen our understanding of estrogen signaling, and have important implications for the design of safer and more effective HRT. PMID- 15780524 TI - Vascular reactivity and atheromatous plaques in post-menopausal women on tibolone treatment. Open prospective study with Doppler ultrasonography in internal carotid artery. AB - BACKGROUND: Arteriosclerosis is the main cause of ischaemic ictus. The middle cerebral and anterior cerebral arteries, which irrigate over 70% of the entire cerebral tissue, spring from the internal carotid. Additionally, it is in the extracraneal vessels that embolism, thrombosis and stenosis originate more frequently. AIM: To evaluate the variations in blood flow (pulsatility index: PI) and to assess the evolution of atherogenic lesions (thickening of the vascular intima and the presence of atheromatous plaques) in the internal carotid artery after tibolone therapy. SUBJECTS AND METHODS: A total of 116 healthy menopausal women were included in this open, prospective and comparative study. Of them, 101 subjects completed the 48 weeks follow-up. Subjects were allocated in two groups: group T (n = 55) received 2.5mg/day of tibolone daily and group C (n = 61) was a free-treatment control group. To evaluate both resistance to blood flow and the existence and evolution of atheromatous plaques in the internal carotid, an ultrasonograph with a pulsed Doppler was used. The PI was used as the parameter of vascular tone. To study atherosclerotic lesions in the internal carotid artery, we used morphological criteria. Measurements were done before entering in the study, and at 12, 24, 36 and 48 weeks of treatment. RESULTS: After tibolone treatment the PI in the internal carotid artery was observed markedly diminished. Moreover, tibolone reduces both the thickness and length of atheromatous plaque and the degree of vascular stenosis. CONCLUSION: tibolone administration reduced the carotid atheromatous plaque in thickness and length and improved cerebral perfusion. PMID- 15780525 TI - Acute administration of 17beta-estradiol improves endothelium-dependent vasodilation in postmenopausal women. AB - OBJECTIVES: Estrogen's effect on endothelial function in postmenopausal women with mild hypertension but no other cardiovascular risk factors remains unclear. This study investigated the effect of an acutely administered therapeutic/low dose of 17beta-estradiol on vasodilation in this patient population. METHODS: Forearm blood flow (FBF) was measured in seven white, hypertensive (blood pressure 144 +/- 8/93 +/- 5 mmHg), postmenopausal (mean age: 54.4 +/- 5 years) women at baseline and during the intra-brachial infusion of increasing doses of acetylcholine (ACh; 0.75, 5, and 15 microg/100 mL tissue/min) and sodium nitroprusside (NP; 1, 2, and 4 microg/100 mL tissue/min). These measurements were obtained both before and after the sublingual administration of 17beta-estradiol. Eight normotensive women (blood pressure 115 +/- 8/76 +/- 5 mmHg) with otherwise similar characteristics served as controls (mean age: 55.8 +/- 5 years). Exclusion criteria included medications or any condition that could alter endothelial function. RESULTS: Before estradiol administration, FBF values at baseline and after all doses of ACh and NP were similar between groups. Acutely administered 17beta-estradiol significantly improved the FBF response to ACh in both the normotensive (maximal response: 17.6 +/- 5 versus 22.5 +/- 7 mL/min/100 mL) and hypertensive (11 +/- 4 versus 16 +/- 6; 12 +/- 4 versus 17 +/- 5 and 14 +/- 3 versus 20 +/- 7 mL/min/100 mL) groups. It also altered the NP dose-response curve in the both groups. CONCLUSION: 17beta-estradiol improved vasodilatory responses in mildly hypertensive postmenopausal women without other risk factors for cardiovascular disease. PMID- 15780526 TI - Effects in post-menopausal women of transdermal estrogen associated with progestin upon the removal from the plasma of a microemulsion that resembles low density lipoprotein (LDL). AB - OBJECTIVE: To evaluate the effects of transdermal estradiol and medroxyprogesterone acetate (MPA) treatment on the removal from the plasma of a cholesterol-rich microemulsion (LDE) that roughly resembles low-density lipoprotein (LDL) structure and that binds to LDL receptors. METHODS: Ten healthy post-menopausal women were studied before and after 3-month treatment with transdermal estradiol in the following dosages administered every 3.5 days: 25, 50, 50, 100, 100, 50, 50 and 25 microg. From the 15th to the 21st day and from the 22nd to the 28th day of estrogen treatment, respectively, 10 and 5 mg q.d. MPA per oral were associated to the transdermal estrogen. The emulsion labeled with 14C-cholesteryl oleate was injected after 12 h fasting and its fractional catabolic rate (FCR) was calculated from the plasma decaying curves of the isotope. RESULTS: Treatment reduced LDL-cholesterol levels by 8% only (149.0 +/- 36.0 mg/dl, 138.0 +/- 27.0 mg/dl; P = 0.046), but the FCR of LDE expressed in medians (25%; 75%) increased from 0.0054 (0.003; 0.052) h(-1) to 0.021 (0.009; 0.10) h(-1), P = 0.002. CONCLUSION: The association used in this study so as to mimic the increasing-decreasing pattern of the hormonal ovarian production reduced modestly LDL-cholesterol levels but pronouncedly increased the lipoprotein removal as tested by LDE FCR. PMID- 15780527 TI - Early impact of hormone replacement therapy on vascular hemodynamics detected via ocular colour Doppler analysis. AB - OBJECTIVE: To determine the effects of hormone replacement therapy (HRT) on ocular blood flow. STUDY DESIGN: In a prospective controlled study, 40 healthy women who presented to the menopause clinic between December 2000 and December 2001 were randomly assigned into the study. The HRT-receiving group was administered estradiol 17-valerate 2 mg the first 11 days, and estradiol 17 valerate 2 mg plus ciproterone acetate 1 mg the next 10 days of the monthly cycle for 6 months. The control group did not receive any HRT for 6 months. The ocular colour Doppler analysis were performed at baseline and after 3 and 6 months. The ocular Doppler analysis was performed in the first half of the cycle in the HRT receiving group. RESULTS: Central retinal artery and ophthalmic artery basal Doppler index (peak systolic velocity, end-diastolic velocity, resistive index and pulsatility index) values of the two groups at the beginning of the study did not show any statistically significant difference. Both the right and the left central retinal artery pulsatility index (PI) values of the study group, who received HRT at the end of the third and sixth months, showed a statistically significant decline (paired-samples test, P < 0.05), while the decrease in the resistive indexes was not significant. CONCLUSION: These results suggest that 6 months of combined hormone replacement therapy with estradiol 17-valerate 2 mg plus ciproterone acetate 1 mg improves ocular vascular Doppler indices which may be a reflection of cerebral vascular status. PMID- 15780528 TI - Effects of transdermal estrogen replacement therapy on plasma levels of nitric oxide and plasma lipids in postmenopausal women. AB - OBJECTIVE: Our purpose was to assess the effects of transdermal estrogen replacement therapy (TERT) on plasma levels of nitric oxide (NO) and plasma lipids in postmenopausal women. MATERIALS AND METHODS: The study designed as a randomized, double-blind, placebo-controlled trial, involved 43 postmenopausal healthy women who had previously undergone hysterectomy. Women received either transdermal 100 microg 17beta-estradiol (Climara forte TTS) or placebo once a week for 3 months. Plasma levels of NO metabolites, estradiol (E2), total cholesterol (TC), triglicerides (TG), low-density lipoproteins (LDL), high density lipoproteins (HDL), HDL2 and HDL3 were measured in blood samples of all women which were collected before, after 24 h and after 3 months of therapy. RESULTS: We found significantly increased NO levels 24 h after therapy in TERT group. Moreover significantly higher NO levels were determined at 3rd month of therapy. Serum HDL and HDL2 levels of ERT group were significantly increased at 3rd month of therapy. Alteration of serum levels of HDL3, LDL and TC were not significantly different in groups. TG levels were significantly decreased in TERT group. DISCUSSION: NO-related mechanism may help to explain the cardio-protective effect of TERT in the postmenopausal period. TERT seems to have favorable effects on plasma lipids in surgical menopausal women. PMID- 15780529 TI - Are dehydroepiandrosterone sulphate and lipids associated with erectile dysfunction? AB - OBJECTIVE: Considering the results of the Massachusetts Male Aging Study (1994) we undertook the current investigation in an attempt to clarify the role of dehydroepiandrosterone sulphate (DHEAS), testosterone and lipids on age related deterioration of erectile function. METHODS: Forty males (13 under 40 years of age and 27 over 40) with erectile dysfunction were investigated. Seventeen healthy subjects (8 under 40 and 9 over 40 years) volunteered as controls. Serum levels of DHEAS, testosterone, total cholesterol, high-density lipoprotein cholesterol (HDL-ch), low-density lipoprotein cholesterol (LDL-ch) and triglycerides were assessed in blood samples. RESULTS: Plasma levels of DHEAS in patients over 40 years of age (4.17 +/- 2.76 micromol/l) were significantly lower in comparison with the younger group of patients (10.49 +/- 3.87 micromol/l), P < 0.001. There was no statistically significant difference in the DHEAS levels between patients and controls in the same age group. DHEAS in the patients showed an inverse correlation with age (r = -0.705, P < 0.001) and a positive correlation with testosterone (r = +0.402, P < 0.01). The same was found in the controls. The HDL-ch results were in the reference range. The total cholesterol levels (5.35 +/- 0.74 mmol/l) and LDL-ch levels (3.58 +/- 0.76 mmol/l) of the patients with erectile dysfunction in the group under 40 years were significantly higher in comparison with the controls (4.21 +/- 0.69 and 2.46 +/- 0.74 mmol/l, respectively, P <0.01). CONCLUSION: The data indicates that the decline in DHEAS is an age-related process rather than a causative factor of erectile dysfunction; total cholesterol and mainly LDL-ch may contribute to erectile dysfunction, especially in younger men. PMID- 15780530 TI - Long-term treatment with vaginal estradiol tablets--an impact on uterine artery blood flow. AB - OBJECTIVE: We investigated, using color Doppler ultrasound, possible detectable blood changes of prolonged treatment with 17-beta-estradiol tablets on uterine artery blood flow, in a cohort of low risk postmenopausal women. METHODS: 39 postmenopausal women, who were taking local estrogen therapy for at least 6 months, were examined in the study group. Forty-two women who never used hormone replacement therapy consisted a control group. They were examined with color Doppler ultrasound and the pulsatility index of the uterine artery was measured. The groups were divided into three subgroups (the age, the duration of the postmenopause, and the duration of the treatment) and the data were compared among groups and within subgroups. RESULTS: The overall mean age of the patients was 66 years, the mean duration of the postmenopause was 15 years, the overall mean PI was 1.96 +/- 0.90, and the mean duration of taking vaginal estradiol tablets was 1.97 years for the study group; and 64 years, 13 years and 2.88 +/- 0.96 for the control group, respectively. There was a difference in the mean PI among groups. No significant differences in pulsatility index values in the subgroups were found. CONCLUSION: Long-term treatments with vaginal estradiol tablets in low risk women causes lowering of the uterine artery PI values compared to the women who are not receiving hormone replacement therapy. PMID- 15780531 TI - Different effect of hormone replacement therapy, DHEAS and tibolone on endothelial function in postmenopausal women with increased cardiovascular risk. AB - Menopause is associated with an increased cardiovascular risk and with a decrease in endothelial function. Hormone replacement therapy (HRT) improves endothelial function in post-menopausal women (PMW) without established atherosclerosis. New alternative treatments, among which tibolone (T) and DHEAS have been suggested to reduce postmenopausal cardiovascular risk. Although, in vitro animal studies have suggested that T and DHEAS improve endothelial function, their effect in humans has never been tested. The aim of the present study was to compare the effects of HRT (continuous combined 0.625 mg conjugated equine estrogen plus 2.5 mg/d medoxyprogesterone) DHEAS and T on endothelium-dependent flow-mediated vasodilatation (FMD), plasma nitrite, nitrate and endothelin-1 levels in 16 PMW with increased cardiovascular risk in a double-blinded, double-crossover study. Women were randomized and treated for 4 weeks with HRT, T or DHEAS. Brachial artery diameter, FMD, endothelin-1 and plasma nitrite and nitrate levels were measured at baseline and after each treatment phase. Brachial artery diameters remained unchanged after each treatment phase. HRT significantly improved FMD compared to both baseline and to T and DHEAS therapies while no effect of T or DHEAS on FMD was noted. In conclusion, HRT, but neither T nor DHEAS, improves endothelial function and reduces plasma levels of endothelin-1 in PMW at risk of CAD. PMID- 15780532 TI - Effect of raloxifene and hormone therapy on serum markers of brain and whole-body cholesterol metabolism in postmenopausal women. AB - OBJECTIVE: To compare the 2-year effects of raloxifene (Rlx) with oral postmenopausal hormone therapy (HT) on serum markers of brain and whole-body cholesterol metabolism. METHODS: In a randomized, double-blind, placebo controlled trial, 95 healthy, non-hysterectomized, early postmenopausal women received either daily Rlx 60 mg (n = 24), Rlx 150 mg (n = 23), HT (conjugated equine estrogens 0.625 mg/medroxyprogesterone acetate 2.5 mg; n = 24), or placebo (n = 24). Fasting blood samples were collected at baseline and after 6, 12, and 24 months of treatment for measurement of serum concentrations of cholesterol by means of gas-liquid chromatography; 24S-hydroxycholesterol (cerebrosterol), lathosterol, and the plant sterol campesterol by means of gas-liquid chromatography-mass spectrometry. The analyses were performed retrospectively from serum samples stored at -70 degrees C for 5 years. RESULTS: Twenty-four months of treatment with raloxifene 150 mg was associated with a significant reduction in serum cholesterol concentrations (-10%, P = 0.007). The ratio of 24S hydroxycholesterol to cholesterol, a serum marker of brain cholesterol metabolism, showed a significant increase after 6 and 12 months with raloxifene 150 mg but not after 24 months (P = 0.001). The ratio of lathosterol to cholesterol, a marker of whole-body cholesterol synthesis, increased with raloxifene 60 mg (P = 0.163), raloxifene 150 mg (P < 0.001), as well as with HT (P = 0.005). The ratio of campesterol to cholesterol, a marker of cholesterol absorption rate, was significantly reduced with HT (P = 0.002). CONCLUSION: Two year treatment with raloxifene or HT had no influence on brain cholesterol metabolism, while whole-body cholesterol synthesis, assessed by the ratio of lathosterol to cholesterol, increased during raloxifene and HT. PMID- 15780534 TI - Polycystic ovary syndrome in post-menopausal women--marker of the metabolic syndrome. AB - OBJECTIVES: To determine among post-menopausal women, the prevalence of polycystic ovary syndrome (PCOS) and to evaluate the endocrine and metabolic profile of these women in comparisons to women without PCOS. METHODS: All women attending two post-menopausal outpatient clinics, fulfilling the inclusion criteria, were evaluated by physical examination, detailed questionnaire and laboratory measurements of glucose, lipids and sex-hormone levels. RESULTS: Among 104 women 7 (6.7%) were diagnosed with PCOS. As compared to women without PCOS, central obesity was more common in the PCOS group (p=0.004), 4/7 had type 2 diabetes compared to 8/97 in the non-PCOS group (p=0.003) and 6/7 versus 31/97 had the characteristic dyslipidemia of the metabolic syndrome (p=0.047). CONCLUSIONS: PCOS is common in post-menopausal women attending outpatient's clinics, and is a marker for a metabolic profile that is associated with a high risk for cardiovascular diseases (CVD). PMID- 15780533 TI - Serum androgen levels and insulin resistance in postmenopausal women: association with hormone therapy, tibolone and raloxifene. AB - OBJECTIVE: To assess endogenous androgen and insulin resistance status in postmenopausal women receiving continuous combined hormone therapy (HT), tibolone, raloxifene or no therapy. METHODS: A total of 427 postmenopausal women aged 42-71 years were studied in a cross-sectional design. Among them 84 were taking HT (46 women conjugated equine estrogens 0.625 mg; medroxyprogesterone acetate, 5 mg, CEE/MPA; and 38 women 17beta-estradiol 2 mg; norethisterone acetate 1 mg, E2/NETA); 83 were taking tibolone 2.5 mg; 50 were taking raloxifene HCl 60 mg; and 210 women were not receiving any therapy. Main outcome measures were FSH, LH, estradiol, total testosterone, SHBG, free androgen index (FAI), Delta4-Androstendione (Delta4-A), Dehydroepiandrosterone sulphate (DHEAS) and HOMA insulin resistance index (HOMA-IR). RESULTS: In women not on hormone therapy smoking and older age was associated with lower DHEAS levels. FAI values increased linearly with increasing BMI. Age and BMI were positive determinants of HOMA-IR, while no association was identified between endogenous sex steroids and insulin resistance. CEE/MPA therapy was associated with higher SHBG, lower FAI and lower HOMA-IR values compared to women not on therapy (age and BMI-adjusted SHBG: CEE/MPA 148.8 nmol/l, controls 58.7 nmol/l, p < 0.01; age-adjusted FAI: CEE/MPA 0.8, controls 3.2, p < 0.05; age-adjusted HOMA-IR: CEE/MPA 1.3, controls 2.6, p < 0.05). On the contrary, E2/NETA treatment had no effect on these parameters. Women on tibolone had lower SHBG, higher FAI and similar HOMA-IR values compared to controls (age and BMI-adjusted SHBG: 24.1 nmol/l, p < 0.01; FAI: 6.0, p < 0.05; HOMA-IR: 2.3, p = NS). Raloxifene users did not exhibit any difference with respect to sex steroids and HOMA-IR levels. CONCLUSIONS: CEE/MPA users had lower free testosterone and improved insulin sensitivity. Tibolone on the other hand associated with higher free testosterone, while raloxifene did not relate to any of these parameters. PMID- 15780535 TI - A comparative study of efficacy of tibolone and simvastatin on atherosclerosis in ovariectomized cholesterol-fed rabbits. AB - BACKGROUND: After menopause women are more susceptible to coronary heart disease due to increased risk of atherosclerosis. Tibolone (Livial) is an innovative synthetic steroid analogue for the treatment of postmenopausal climacteric symptoms including atherosclerosis, but the mechanisms of its effect are still unclear. The present study investigated the effect of tibolone and simvastatin on atherosclerosis and the expression of both estrogen receptor A (ERA) and LDL receptor (LDLR) mRNA in ovariectomized cholesterol-fed rabbits. METHODS: Fifty New Zealand white rabbits were included for the study. Of them, 40 underwent bilateral ovariectomy and the other 10 were sham-operated. The sham-operated group only received atherogenic diet (group SC) and the ovariectomized rabbits were divided into 4 groups of 10 each, with group N received normal diet, group C received atherogenic diet, group T received atherogenic diet and tibolone (2.5 mg/day) and group SI received atherogenic diet and simvastatin (20 mg/day). After 12 weeks of the treatments, the animals were euthanized and the extent of thoracic aortic atherosclerosis was measured morphologically and the level of ERA and LDLR mRNA in heart and liver was determined by real-time RT-PCR. RESULTS: The extent of atherosclerosis in the thoracic aorta was 0.75+/-0.24 for group C, 0.56+/-0.27 for group SC, almost 0 for group N, 0.10+/-0.02 for group T and 0.09 +/-0.08 for group SI (P<0.01; groups T versus C, T versus SC, SI versus C, SI versus SC). The relative copies of ERA at group C, SC, N, T and SI were 0.29, 0.53, 0.46, 0.85 and 0.30, respectively in heart and 0.32, 0.51, 0.49, 0.68 and 0.30, respectively in liver; the relative copies of LDLR at group C, SC, N, T and SI were 0.22, 0.24, 0.33, 0.27 and 0.23, respectively in heart and 0.68, 0.93, 1.52, 1.27 and 0.88, respectively in liver. CONCLUSION: Both tibolone and simvastatin prevented the atherosclerosis in ovariectomized cholesterol-fed rabbits and this effect was associated with up-regulation of ERA and LDLR expression by tibolone but not by simvastatin. PMID- 15780536 TI - Lipids and clotting factors during low dose transdermal estradiol/norethisterone use. AB - OBJECTIVE: To demonstrate the effects of 2-year transdermal continuous combined low-dose estradiol (0.025 mg/day) and norethisterone acetate (0.125 mg/day) on lipid/lipoprotein profile and coagulation/fibrinolysis. METHODS: A double-blind, randomized, multicenter, parallel, 1-year trial enrolled 266 healthy women at least 2 years post menopause. Patients received either 0.025 mg estradiol and 0.125 mg norethisterone acetate daily or placebo transdermally. One hundred and thirty five women completed a second year open follow-up (96 had used Estragest TTS, 39 placebo during the first year), where all women had the estradiol/norethisterone patch. Lipid/lipoprotein profile and coagulation/fibrinolysis parameters were studied at 0, 24, 48, 72 and 96 weeks. RESULTS: In women on estradiol/norethisterone total cholesterol, Lp(a) and VLDL cholesterol decreased significantly more than in the placebo group after 24 weeks and LDL cholesterol after 48 weeks. Women on estradiol/norethisterone had no change in HDL, triglycerides or Lp(a), an increased HDL/total cholesterol ratio and decreased LDL, VLDL and total cholesterol at 48 weeks compared to placebo. Women with active treatment also showed a significant reduction compared with the placebo group of Factor VII and antithrombin III at 24 and 48 weeks and a reduction of fibrinogen at 24 weeks. These changes persisted over the second year. CONCLUSIONS: A continuous combined low-dose transdermal patch daily delivering 0.025 mg estradiol and 0.125 mg norethisterone acetate provided beneficial effects on lipid/lipoprotein profile and coagulation/fibrinolysis. The changes were similar to those previously described after higher dose oral and transdermal estrogen/progestogen regimens. PMID- 15780537 TI - Intranasal 17beta-estradiol treatment and Vitamin B12, folate and homocysteine in menopause. AB - OBJECTIVE: This study assessed the effect of intranasal administration of 17beta estradiol (Aerodiol) on plasma levels of homocysteine, Vitamin B12 and folate in postmenopausal women. METHODS: In all, 26 symptomatic postmenopausal women who had undergone hysterectomy and oophorectomy at least 12 months previously participated in this 6-month randomized prospective clinical study. Menopause was determined by serum FSH level >30 microIU/ml and serum estradiol concentration <30 pg/ml. Intranasal 17beta-estradiol treatment was given once daily at a standard daily dose of 300 microg to 16 women, and 10 did not receive any treatment. RESULTS: In the group receiving intranasal 17beta-estradiol, mean (+/ S.D.) plasma homocysteine level decreased significantly from pre-treatment values (from 16.68+/-4.33 to 14.15+/-1.18 nmol/ml, p=0.029) and the mean folate level increased (from 4.11+/-0.80 to 5.64+/-1.87 ng/ml, p=0.012). Vitamin B12 levels showed a tendency towards increasing. In the treated group, significant negative correlations were observed between homocysteine and folate values (r=-0.586, p=0.017) and between homocysteine and Vitamin B12 values (r=-0.672, p=0.004). No significant changes were observed in the untreated group. CONCLUSION: The reduction in plasma homocysteine levels observed after 6 months' treatment with intranasal 17beta-estradiol may reflect an alteration in folate and Vitamin B12 homeostasis. PMID- 15780539 TI - Women, Co-occurring Disorders, and Violence Study: evaluation design and study population. AB - The Women, Co-occurring Disorders, and Violence Study (WCDVS) was a multi-site cooperative study to evaluate new service models for women with co-occurring mental health and substance use disorders and a history of physical and/or sexual abuse. Despite common features in the service interventions and evaluation procedures, diversity across the nine sites plus differences introduced by non random assignment led to numerous methodological challenges. This article describes the design, measurement, and analysis decisions behind the WCDVS and lays the foundation for understanding participant-level outcomes and service costs. This article also describes the study population, as recruited and following attrition at the 6-month follow-up, in order to address the threat of selection bias to inferences drawn from this multi-site study. PMID- 15780540 TI - Outcomes for women with co-occurring disorders and trauma: program-level effects. AB - Program-level effects at 6 months are reported from meta-analysis of a nine-site quasi-experimental study of comprehensive, integrated, trauma-informed, and consumer-involved services for women who have mental health problems, substance use disorders, and who have experienced interpersonal violence. The average weighted effect size is significant for the treatment condition for improved post traumatic symptoms (p < 0.02), drug use problem severity (p < 0.02), and nearly significant for mental health symptoms (p < 0.06). There is significant heterogeneity in effect sizes across sites. Program-level variables were examined in an effort to explain this heterogeneity. The findings indicate that sites which provided significantly more integrated counseling produced more favorable results in mental health symptoms (p < 0.01) and both alcohol (p < 0.001) and drug use problem severity (p < 0.001). The same trend is observable for reductions in post-traumatic stress symptoms, although the difference does not attain statistical significance. PMID- 15780541 TI - Outcomes for women with co-occurring disorders and trauma: program and person level effects. AB - Six-month outcomes are evaluated from a 9-site quasi-experimental study of women with mental health and substance use disorders who have experienced physical or sexual abuse who enrolled in either comprehensive, integrated, trauma-informed, and consumer/survivor/recovering person-involved services (N = 1023) or usual care (N = 983). Mental health, post-traumatic stress symptoms, and substance use outcomes are assessed with multilevel regression models, controlling for program and personal characteristics. Person-level variables predict outcomes independent of intervention condition and, to a small extent, moderate intervention and program effects. In sites where the intervention condition provided more integrated counseling than the comparison condition, there are increased effects on mental health and substance use outcomes; these effects are partially mediated by person-level variables. These results encourage further research to identify the longer-term effects of integrated counseling for women with co-occurring disorders and trauma histories. PMID- 15780542 TI - Service costs for women with co-occurring disorders and trauma. AB - Several aspects of costs related to health care and other service use at 6-month follow-up are presented for women with co-occurring mental health and substance abuse disorders with histories of physical and/or sexual abuse receiving comprehensive, integrated, trauma-informed and consumer/survivor/recovering person-involved interventions (n = 1023) or usual care (n = 983) in a nine-site quasi-experimental study. Results show that, controlling for pre-baseline use, there are no significant differences in total costs between participants in the intervention condition and those in the usual care comparison condition, either from a governmental (avg. US dollars 13,500) or Medicaid reimbursement perspectives (avg. just over US dollars 10,000). When combined with clinical outcomes analyzed in other works in this issue by Cocozza et al. (2005) and Morrissey et al. (2005), which favored the intervention sites, these cost findings indicate that the treatment intervention services are cost-effective as compared with the usual care received by women at the comparison sites. PMID- 15780543 TI - Women, Co-occurring Disorders, and Violence Study: a case for trauma-informed care. PMID- 15780544 TI - Improving the quality of demonstration research: Integrating those with personal experience. PMID- 15780545 TI - Outcomes for clients of public substance abuse treatment programs before and after Medicaid managed care. AB - Following conversion of Medicaid substance abuse treatment programs in Oregon from fee-for-service to managed care, there were suggestions of poorer outcomes. This project interviewed cohorts of Medicaid and non-Medicaid substance abuse treatment clients before and after the 1995 implementation of Oregon's Medicaid capitated payment system. Subjects were 553 adults (59% male, average age 35, 64% white). Forty-three percent were Medicaid clients and 60% were treated during the fee-for-service era. All clients were interviewed with the Addiction Severity Index at baseline and at 6 and 12 months thereafter. Multilevel, multivariate analyses examined baseline and outcome differences between the Medicaid and Non Medicaid groups; between the fee-for-service and managed care conditions; and their interactions. After adjustment for covariates, most analyses were not statistically significant. It does not appear that Medicaid managed care had an adverse impact on outcomes for clients with substance abuse problems. PMID- 15780546 TI - The Drinker's Check-up: 12-month outcomes of a controlled clinical trial of a stand-alone software program for problem drinkers. AB - Sixty-one problem drinkers were randomly assigned to either immediate treatment or a 4-week wait-list control group. Treatment consisted of a computer-based brief motivational intervention, the Drinker's Check-up (DCU). Outcomes strongly support the experimental hypotheses and long-term effectiveness of the treatment. Overall, participants reduced the quantity and frequency of drinking by 50%, and had similar reductions in alcohol-related problems that were sustained through 12 month follow-up. The DCU seems to be effective in enhancing problem drinkers' motivation for change. PMID- 15780547 TI - Development and initial demonstration of a community-based intervention for homeless, cocaine-using, African-American Women. AB - Drug abstinence is difficult to achieve and maintain, especially when clients return to their pretreatment environment. Forging ties with the natural helping networks in the community, such as religious organizations, may reinforce abstinent behaviors established during treatment. This study evaluated Bridges to the Community, a supplemental component to an intensive residential treatment program. Bridges uses members of African-American churches as mentors for recovering women. This demonstration project included 118 female participants with primary cocaine dependence who received either standard treatment or Bridges plus standard treatment. Participants in both groups reduced substance use, risk taking behaviors, depression, and increased self-esteem. Participants who received Bridges had greater treatment retention, reported 100% cocaine abstinence at follow-up, and were more satisfied with their treatment. PMID- 15780548 TI - Knowledge of and interest in hepatitis C treatment at a methadone clinic. AB - This study describes knowledge about hepatitis C virus (HCV) infection and interest in treatment among 110 opiate dependent patients from an opiate dependence treatment program in San Francisco. None had been treated for HCV and only 30% had been evaluated for HCV treatment. While only 34% knew about HCV treatment, 54% of the sample became "definitely interested" in HCV treatment after hearing the risks and benefits. Men were approximately five times more likely than women to know of some HCV treatment. Whites were seven times and Latinos were about six times more likely than African-Americans to know about HCV treatment. Our findings suggest that methadone programs can play an important role in increasing access to HCV treatment through educating patients about treatment options. PMID- 15780549 TI - The role of depression symptoms in predicting drug abstinence in outpatient substance abuse treatment. AB - OBJECTIVE: This study examined the role of depressive symptoms in the context of specific demographic and individual treatment characteristics in predicting drug abstinence at discharge from outpatient substance abuse treatment. METHODS: Data from 827 clients entering a large public funded outpatient substance abuse treatment program were analyzed using logistic regression to assess the effects of depressive symptoms on drug abstinence status at discharge. Analyses on the effects of gender, race, age, education level, frequency of drug use, insurance status, referral source, and length of stay in treatment on drug abstinence status at discharge were also conducted. RESULTS: Higher depressive symptom scores significantly predicted a decreased likelihood of clients' abstinence at discharge even after accounting for other significant demographic and treatment variables such as insurance status, race, age, primary drug of choice, frequency of drug use at admission and length of stay in treatment. CONCLUSION: The findings suggest that depression symptoms are an important factor affecting successful substance abuse treatment outcomes. Treatment approaches that address depressive symptoms are likely to enhance substance abuse treatment outcomes in real world clinical settings. PMID- 15780550 TI - A controlled trial of amlodipine for cocaine dependence: a negative report. AB - Preclinical models of cocaine dependence have shown favorable reductions in cocaine use using dihydropyridine calcium channel antagonists. This is one of the first reports testing the efficacy of the long-acting calcium channel antagonist, amlodipine, for the treatment of cocaine dependence. This was a 12-week, double blind, randomized, placebo-controlled, parallel patient group trial of amlodipine vs. placebo for the treatment of cocaine dependence. One hundred and sixteen subjects participated in a 12-week medication trial in which 60 subjects received medication and 56 received placebo. Subjects in both groups received up to 12 standard manual-driven cognitive behavioral therapy sessions. Overall, drop-out rate for both groups was high, with only about 20% of subjects completing all 12 weeks of treatment. Both groups showed comparable levels of medication compliance and therapy attendance. In the end, amlodipine was no more effective than placebo in reducing craving or measured levels of cocaine use. PMID- 15780551 TI - Civil legal needs of individuals in drug treatment. AB - This report describes the findings of a pilot study that examined 17 domains of civil legal problems among individuals in drug treatment in one urban setting, the extent to which they seek help to resolve a potential legal problem, and their reasons for not seeking legal assistance. More than half the participants reported at least one legal problem related to health care and/or work, and nearly half reported legal problems related to their family, civil liberties, and/or neighborhood/community concerns. Less than one in five, however, sought legal assistance for problems in the five top-ranked domains, with the exception of family problems. The primary reasons for not seeking assistance were a lack of awareness that a problem was a legal issue and/or uncertainty about who could help. While preliminary in nature, the pilot study suggests that a significant proportion of treatment clients have civil legal problems and that treatment programs could help clients identify legal problems and resources for assistance. PMID- 15780553 TI - Thrombomodulin: tumour biology and prognostic implications. AB - BACKGROUND: Thrombomodulin (TM) is an endothelial receptor that exerts anti coagulant, anti-fibrinolytic, and anti-inflammatory activity by inhibiting thrombin and cellular adhesion. There is growing evidence that TM plays a role in tumour behaviour. METHODS: The electronic literature (1966-2004) was reviewed with a specific focus on tumour biology. RESULTS: TM is expressed on both the endothelium and tumour cells in several cancers. Loss of expression denotes a more malignant profile with poorer prognosis. Loss of TM is mediated by hypoxia, endotoxin, and various cytokines, while up-regulation can be achieved by pharmacological manipulation (e.g. pentoxyfylline and statins). CONCLUSION: Originally described as an endothelial anticoagulant, TM plays a key role in tumour biology and prognostics, and provides a potential therapeutic target in impeding cancer spread. PMID- 15780554 TI - Incidence of axillary recurrence in 113 sentinel node negative breast cancer patients: a 3-year follow-up study. AB - BACKGROUNDS/AIMS: This study evaluates the 3-year follow-up period and recurrence rate in patients with a negative sentinel node biopsy (SNB) without an additional axillary dissection (ALND). METHODS: Between January 2000 and March 2002, 197 patients with an invasive breast cancer and clinically negative axillary nodes underwent a sentinel node biopsy. One hundred and thirteen patients were included in our study. The follow-up consisted of clinical examination every 3 months in the first year, followed by every 6 months after the first year. A mammography was obtained annually. Attention was paid to loco-regional recurrence, including axillary recurrence, and distant metastases. RESULTS: The mean duration of follow up was 37.5 months (range 24-54). In this period, one patient was diagnosed with an axillary recurrence and one patient developed a supraclavicular lymph node metastasis. Two patients developed a second primary breast cancer in the contralateral breast. No patients were diagnosed with distant metastasis. CONCLUSION: These 3 year follow-up results suggest that SNB is a procedure with a low clinical recurrence rate, which can replace, when strict criteria are met, ALND if the sentinel node is negative. PMID- 15780555 TI - Axillary recurrence in breast cancer. AB - AIM: To determine whether axillary recurrence reflects inadequate axillary treatment or adverse pathological features. METHODS: The case-records were reviewed of 2122 women aged under 75 years, treated for invasive breast cancer during the time-period 1/1/86-31/12/91 in a geographically defined area. Data were abstracted on operations performed, pathological features, post-operative treatments and details of axillary recurrence. The risk of axillary recurrence was examined by pathological, treatment and patient factors. RESULTS: Axillary recurrence was more than twice as likely after inadequate compared to adequate treatment of the axilla (adequate staging or axillary radiotherapy or clearance). Delayed treatment of the axilla was not as successful as adequate primary treatment: multiple axillary recurrences were twice as common, one third of which were uncontrolled at time of death. Inadequate surgical treatment was associated with increased rates of recurrence despite endocrine therapy, chemotherapy or radiotherapy. Lymphoedema was twice as common if axillary radiotherapy was combined with any axillary surgical procedure. CONCLUSIONS: Axillary recurrence is more common in tumours with adverse pathology but may also result from inadequate axillary treatment. In order to minimise axillary recurrence, optimal treatment of the axilla entails adequate staging (sampling of four or more nodes) and treatment (axillary clearance or radiotherapy and endocrine therapy) in all women. PMID- 15780556 TI - Breast cancer diagnosed during pregnancy and lactation: biological features and treatment options. AB - AIM OF THE STUDY: Assessment of biological features and treatment of patients with breast cancer presenting during pregnancy or lactation. PATIENTS AND METHODS: Immunohistochemical analysis of estrogen receptor (ER) and progesterone receptor (PgR), Ki-67, HER2/neu, prognostic markers, treatment and follow-up of 21 patients with breast cancer during pregnancy (BCdP) and 17 with breast cancer during lactation (BCdL) are presented. RESULTS: Median age was 36 and 33 years, median tumour size was 2.4 and 2.5 cm, axillary lymph nodes were positive in 10 of 21 pregnant patients and 11 of 17 lactating patients, respectively. Both ER and PgR were not expressed in six of 21 pregnant women and nine of 17 lactating patients. All the six women who had concurrent diagnosis of breast cancer and pregnancy (first trimester) preferred termination of pregnancy although an alternative option was discussed. Five patients received anthracycline containing chemotherapy during the second and third trimester with no complications for patient and child. Conservative surgery was performed in 15 of 21 patients during pregnancy with no local reappearance after a median follow-up of 24 months. Three pregnant women underwent lymphoscintigraphy and sentinel lymph node biopsy. CONCLUSIONS: Patients who had concurrent diagnosis of breast cancer and pregnancy (early first trimester) preferred termination of pregnancy to allow easier completion of treatment. Conservative surgery was safe also in women with BCdP. Sentinel node biopsy might be considered for pregnant patients with a clinically negative axilla. PMID- 15780557 TI - The assessment of urinary function following extended lymph node dissection for colorectal cancer. AB - AIMS: The aim of the current study is to demonstrate whether the effects of extended systematic lymph-node dissection (ESLND) on urinary dysfunctions differ from those of curative radical surgery (CRS) only for rectal cancer. METHODS: We present data about our patients who underwent rectal resection for rectal cancer over 5 years. One hundred and seventy patients with rectal cancer were reviewed with respect to surgical procedures and post-operative urinary problems. RESULTS: We performed CRS on 146 patients and CRS+ESLND on 24 patients, and analysed the incidence of post-operative urinary dysfunction in both groups. Urinary incontinence rates were 39 and 58%, urinary retention rates were 4 and 16%, for the patients from CRS group and CRS+ESLND group, respectively. CONCLUSIONS: We conclude that the addition of ESLND to CRS does not increase the frequency of post-operative urinary dysfunction, apart from an increased risk of urinary retention. PMID- 15780558 TI - Changes in outcome with sphincter preserving surgery for rectal cancer in Korea, 1991-2000. AB - AIM: To report the clinical and oncological data of patients operated on for rectal cancers 3-5 cm from the AV over a 10 year period, including the Sphincter preservation (SP) rate. METHODS: We reviewed medical records of 304 patients with rectal cancers 3-5 cm from the AV who underwent surgical resection from January 1991 through December 2000. The 10 years were divided into three periods based on the introduction of new surgical techniques, specifically, ultralow anterior resection (ULAR) with double stapling in March 1994 and ULAR with coloanal anastomosis in April 1997. The rates of SP, complications and patient survival during these periods were compared. RESULTS: The SP rate increased significantly over the 10 years, from 16.4% in period I (January 1991-February 1994), to 53.0% in period II (March 1994-March 1997), to 86.5% in period III (April 1997-December 2000) (p<0.001). Over time, the age of the patients increased (p=0.004), the length of the distal resection margin became shorter (p=0.005), and the rate of lymph node metastasis increased (p=0.016). The factors significantly influencing SP were the period (p<0.001) and the distance from the AV (p<0.001). Over time, morbidity did not increase, and overall and disease free survival rates did not decrease. In contrast, the overall survival of N2 cases significantly increased over time (p=0.0492). CONCLUSION: Over 10 years, the SP rate in rectal cancers 3 5 cm from the AV was significantly increased by the introduction of the double stapling and coloanal anastomosis techniques. These surgical methods, however, had no effect on morbidity, disease free survival and overall survival rates. PMID- 15780559 TI - Anorectal reconstruction by electrostimulated graciloplasty as part of abdominoperineal resection. AB - AIMS: To report the outcomes of anorectal reconstruction by electrostimulated graciloplasty as part of abdominoperineal resection, on data prospectively collected over 10 years. PATIENTS AND METHODS: Twenty-three abdominoperineal resections were associated to coloperineal pull through, double graciloplasty and loop abdominal stoma. Fifteen patients also received an implantable pulse generator, either for unsatisfactory result after external-source intermittent electrostimulation and biofeedback (five cases) or during graciloplasty (10 cases). Follow-up was to a maximum of 10 years. Functional outcome was followed up in sixteen patients who underwent stoma takedown. RESULTS: Mean actuarial survival at 5 years was 72.3%. Satisfactory results (score < or =8) occurred in 75% of patients (three without and 13 with stimulator) in the early stages, decreasing to 57% at 1 year and gradually increasing up to 100% at 5 years and over. CONCLUSIONS: Total anorectal reconstruction yields a good functional outcome over time. Thus, despite, and because of, a high complication rate and a great drain on resources, it should be considered a suitable procedure only for selected, strongly motivated patients. PMID- 15780560 TI - CDH1 associated gastric cancer: a report of a family and review of the literature. AB - AIMS: Hereditary diffuse gastric cancer (HDGC) is an autosomal-dominant inherited form of gastric cancer associated with inactivating germline mutations in the CDH1 gene. We set out to outline the role of CDH1 in HDGC. Investigation of a family suspected as having HDGC is discussed. The role of surgery in the management of affected individuals is then examined. METHODS: A search was conducted of Medline and the National Library of Medicine to identify key articles concerning CDH1 gene mutations, familial gastric cancer and gastrectomy. Further, relevant articles were obtained by manual scanning of the reference lists of identified papers. Mutation-specific CDH1 genetic testing was performed on six living family members and on gastric tissue obtained from two deceased members. RESULTS: CDH1 mutations cause inactivation of the cell adhesion protein E-cadherin. Carriers of the CDH1 germline gene mutation develop an aggressive, diffuse, submucosal gastric cancer at an early age. Current endoscopic screening is ineffective at detecting HDGC. The presence of a CDH1 germline gene mutation was confirmed in both deceased family members and also in four of the six living members tested. CONCLUSION: Genetic counselling and CDH1 gene mutation testing is indicated in families with suspected HDGC. In the absence of a satisfactory surveillance mechanism, prophylactic total gastrectomy would appear to be an appropriate therapeutic option in mutation carriers. PMID- 15780561 TI - The role of follow-up endoscopy after total gastrectomy for gastric cancer. AB - BACKGROUND: Follow-up endoscopy after total gastrectomy for gastric cancer is commonly performed without there being any definite evidence of clinical relevance. Therefore, we investigated the role of the upper endoscopic examinations after total gastrectomy for gastric cancer. METHODS: The medical records of 212 early gastric cancer (EGC) patients and 622 advanced gastric cancer (AGC) patients who underwent follow-up endoscopic examination after total gastrectomy between 1994 and 2001 were reviewed. RESULTS: Two of 212 EGC patients and 233 of 622 AGC patients revealed tumour recurrence at all sites. All the endoscopically accessible local tumour recurrences (n=24) were found in the AGC group. Anastomosis site stenosis was detected in 72 of 834 patients. CONCLUSION: Follow-up endoscopy after total gastrectomy for gastric cancer is useful in detecting complications and tumour recurrence. However, this procedure has a limited role in the clinical management and overall survival for patients with recurrent gastric adenocarcinoma. PMID- 15780562 TI - Prognostic value of nodal micrometastases in patients with cancer of the gastro oesophageal junction. AB - AIMS: Aim of this study was to examine the presence and the prognostic impact of immunohistochemically identified nodal micrometastases in patients with gastro oesophageal junction (GEJ) carcinomas. METHODS: Between January 1988 and December 2000, 148 patients underwent a radical (R0) resection with a two-field lymphadenectomy for a GEJ carcinoma. Specimens of 60 patients in whom conventional haematoxylin and eosin (H & E) examination did not demonstrate lymph node metastases (pN0) were available for immunohistochemical (IHC) analysis using antibodies AE1/AE3 directed against cytokeratins. Paraffin embedded material of all retrieved lymph nodes in these patients were serially sectioned and analysed by one pathologist after H & E examination for the presence of micrometastases by IHC. RESULTS: In 60 resection specimens initially staged as pN0 a total of 524 lymph nodes were available for IHC analyses. Micrometastases were detected in 126 out of 524 lymph nodes (24%), corresponding with 18 of the 60 patients (30%) who were upstaged by this technique. Compared with the pN0 group, the disease free survival (DFS) was significantly lower in patients with nodal involvement at IHC (p<0.001). Survival of patients with IHC identified micrometastatic disease was comparable to those with H & E positive lymph nodes. CONCLUSIONS: Micrometastases in regional nodes were detected by cytokeratin-specific IHC in 30% of radical resected GEJ tumours without overt nodal involvement. Their presence conveys a worse prognosis with a significant reduced DFS, suggesting that the finding of micrometastases should be included in the staging system. PMID- 15780563 TI - Recurrent laryngeal nerve paralysis (RLNP) following esophagectomy for carcinoma. AB - BACKGROUND: The aim of this study was to report the frequency of post-operative recurrent laryngeal nerve paralysis (RLNP) following resection for esophageal carcinoma. PATIENTS AND METHODS: Four hundred and four patients were studied. Diagnosis of post-operative RLNP was performed by indirect laryngoscopy. Tumour characteristics, surgical approach and perioperative morbidity and mortality following esophageal resection were recorded. RESULTS: Sixty patients were diagnosed with post-operative RLNP, of whom 47 had a unilateral and 16 a bilateral lesion. RLNP was more frequently diagnosed after transhiatal resection with cervical esophagogastrostomy as compared to abdomino-thoracic resection (p=0.06). A higher rate of post-operative pneumonia was evident in patients with RLNP (33 of 63 as opposed to 90 of 341; p=0.027). CONCLUSION: RLNP is associated with a significant morbidity, especially pulmonary complications after resection of esophageal cancer. PMID- 15780564 TI - Cystic neoplasms of the pancreas with mucin-production. AB - AIM: To compare the clinico-pathological features of intraductal papillary mucinous cystic tumours (IPMT) and mucinous cystic tumours (MCT) of the pancreas. METHODS: Eighteen patients with IPMT and 18 with MCT who underwent surgical resection between 1990 and 2004 were retrospectively reviewed. Their clinico pathological features were compared using univariate analysis. Statistical analyses of potential predictive factors of malignancy for each of these two groups were also conducted. RESULTS: Patients with IPMT were found to be older (64+/-10 vs 43+/-18 years, p<0.001) and were predominantly male (male:female ratio, 5:4 vs 1:17, p=0.003) as compared to patients with MCT. MCTs were found in the body-tail region (100%) whereas IPMTs were more evenly distributed (50% in the head) (p=0.001). Pathologically, IPMT was distinct from MCT in terms of size (3.8+/-3.2 vs 9.1+/-4.4 cm, p=0.001), association with secondary pancreatitis (50 vs 0%, p=0.011), communication with the pancreatic duct (94 vs 0%, p<0.001), presence of a dilated main pancreatic duct (61 vs 0%, p<0.001) and the presence of ovarian-type stroma (0 vs 44%, p=0.003). CONCLUSION: IPMT and MCT are distinct clinico-pathological entities. This distinction is important as management and outcome of these entities may differ. PMID- 15780565 TI - Usefulness of fine needle aspiration cytology in the diagnosis of loco-regional recurrence of differentiated thyroid carcinoma. AB - AIM: The aim of our study was to define the usefulness of fine needle aspiration cytology (FNAC) in the assessment of loco-regional recurrence of differentiated thyroid carcinoma (DTC). METHODS: Among 1182 consecutive patients treated and followed for DTC from 1992 to 2001, we retrospectively analysed 65 FNAC results of patients presenting a suspicion of loco-regional recurrence. Recurrences were proved at histology in 35 cases and by cervical radioiodine uptake on post therapeutic WBS (whole body scan) in nine cases. RESULTS: Among the 44 recurrences, FNAC results were malignant, benign and unsatisfactory in 33, two and nine cases, respectively. For the diagnosis of malignancy, FNAC sensitivity was 94%, specificity 100%, positive predictive value 100% and negative predictive value 87%. In the 35 cases where divergent results between diagnostic WBS (37 111MBq (131)I) and Tg level were observed, FNAC assessed the final status in 22 cases (malignant and benign in 17 and five cases, respectively). Of the 12 non functioning and non-secreting lesions, FNAC diagnosed malignancy in four of the five malignant cases and ruled out malignancy in all seven benign lesions. CONCLUSION: These results outline the interest of FNAC in the assessment of loco regional recurrences of DTC, especially when classical follow-up tools such as WBS and/or Tg level are unable to detect the recurrences. PMID- 15780566 TI - Modification of the infra hyoid musculo-cutaneous flap. AB - BACKGROUND: In 1986, Wang described the infra hyoid musculo-cutaneous flap. Here, we report technical adaptations and improvements to this flap. METHODS: From 1994 to 1996, we performed 61 infrahyoid flaps using the classical procedure. After 5 years of technical evolution, we studied a new series of 91 flaps from January 2000 to June 2002. We reviewed our experience with the infra hyoid flap and described the surgical procedure, its evolution and the impact on the viability of the flap. RESULTS: In the two series, the main arterial pedicle was usually the superior thyroid artery. Venous drainage was more variable; consequently we always performed a modified neck dissection with preservation of the internal jugular vein. In the first series, the surgical results were good with only seven local complications essentially skin necrosis. The functional and aesthetic results seemed acceptable. In the second series, the results, after technical evolution with better venous drainage and cervical closure, were improved (only one necrosis) and the indications were extended. CONCLUSION: With experience, the infra hyoid myocutaneous flaps is reliable and appears as a particularly useful flap for oral cavity, oral pharynx and pharyngeolaryngeal reconstruction. PMID- 15780568 TI - The role of urinary cytology for detection of bladder cancer. AB - PURPOSE: The aim of the present study was to test the value of urinary cytology in the diagnosis of bladder cancer. MATERIALS AND METHODS: One thousand three hundred and eighty voided urine and bladder wash specimens of 495 patients were evaluated by urinary cytology. All patients then underwent transurethral resection of suspicious bladder areas if cystoscopy and/or preceding biopsy were positive. Statistical differences were analysed using the two-sided Fisher's exact test and Cochran's test (p<0.05). RESULTS: In 495 patients including 142 patients with bladder cancer urinary cytology revealed a sensitivity of 38.0% and a specificity of 98.3% with a positive and negative predictive value of 90.6 and 78.6, respectively. Sensitivity increased significantly with malignancy grade (p<0.05). In high grade tumours sensitivity improved from initial 52.2% up to 78.3% after the third sample. In sensitivity and specificity of voided urine and barbotage washing samples no significant difference was detected. CONCLUSIONS: Urinary cytology has its place as an additive diagnostic tool to cystoscopy. None of the currently available urinary markers can replace cystoscopy but are helpful for specific diagnostic problems. PMID- 15780567 TI - S100A1 and KIT gene expressions in common subtypes of renal tumours. AB - AIM: The aim of this study is to evaluate the S100A1 and KIT as gene markers for the differentiation of common subtypes of renal tumours. METHODS: Fifty-five tissue samples (15 clear cell RCCs, 15 papillary RCCs, 7 chromophobe RCCs, 8 oncocytomas and 10 normal renal tissues) were studied The gene expressions of S100A1 and KIT were analysed by one-step RT-PCR by using the specific primers. RESULTS: S100A1 was expressed in 2/15 clear cell RCCs, 11/15 papillary RCCs, 7/8 oncocytomas and in 0/7 chromophobe RCCs. KIT gene was expressed in 6/7 chromophobe RCCs and 7/8 oncocytomas while 0/15 clear cell RCCs and 1/15 papillary RCCs expressed kit gene. Normal tissue expressed neither S100A1 nor KIT gene. CONCLUSION: S100A1 and KIT can be used as gene markers for the differentiation of common subtypes of renal tumours. PMID- 15780570 TI - Improved survival with VATS pleurectomy-decortication in advanced malignant mesothelioma. AB - AIMS: Malignant mesothelioma is increasing in incidence and no current therapy significantly prolongs survival. Previous surgical strategies involved high-risk open procedures without achieving histologically clear resection margins. We present the results of VATS debulking pleurectomy-decortication in advanced disease. METHODS: A consecutive series of patients with suspected malignant mesothelioma underwent thoracoscopic assessment to determine the feasibility of decortication, where this was not possible a biopsy alone was taken. Post operative radiotherapy was administered to port sites, but no other adjuvant therapy was given. The two groups (biopsy only and pleurectomy-decortication) were composed of patients with histologically confirmed mesothelioma [28 and 51 patients, respectively]. The primary endpoint was comparison of actuarial patient survival. Secondary endpoints included post-operative air leak and duration of hospital stay. RESULTS: The overall actuarial survival was 288 days and 67% of patients had died at the time of data analysis. The groups were matched for patient and tumour-related characteristics including age (66, 64 years, p=0.39) and tumour stage (median IMIG stage 3 [IQR 2-3] both groups, p=0.54). The biopsy only group had fewer air leaks (57, 84%, p=0.01) and a shorter hospital stay (4, 8 days, p=0.03). However, the pleurectomy-decortication group had favourable actuarial survival relative to the biopsy only group (416, 127 days, p<0.001). Multivariate analysis showed early stage (p<0.001), absence of pre-operative fever (p=0.03) and pleurectomy-decortication (p<0.001) as independent predictors of survival. CONCLUSION: VATS pleurectomy-decortication is feasible in the majority of cases and independently improves survival for patients with advanced malignant mesothelioma. PMID- 15780569 TI - Prediction by quantitative histology of pathological stage in prostate cancer. AB - AIMS: To find a predictor of extraprostatic extension in clinically localized prostate cancer (PCa), pre-operative ultrasound-guided prostate needle biopsies and clinico-pathological data were reviewed. METHODS: One hundred and eighty three consecutive patients who underwent radical retropubic prostatectomy for clinical T1-T2 PCa and serum PSA <10 ng/ml were reviewed. Pre-operative biopsy was performed according to an extended protocol and whole-mount prostatectomy specimens were processed. The following biopsy variables were categorized to this analysis: Gleason score (< or =6, >6), TPC (< or =20%; >20%), GPC (< or =50%; >50%), cancer-positive cores (< or =2; >2), cancer-positive cores in both lateral portions (yes; no), PCa (monolateral; bilateral). RESULTS: Only 60/183 specimens showed an organ-confined PCa; the remaining ones showed pT3a in 57 cases, pT3b in 11 and pT3 with positive surgical margins in 55. A locally advanced PCa was found in 60.2 and 76.8% of T1c and T2 clinical stage, respectively. The positive predictive value and negative predictive value of biopsy findings to predict a locally advanced PCa was 89.9 and 75%, respectively. All biopsy variables associations were statistically significant; however, among these variables (non categorized), in multivariate logistic regression analysis, only GPC was significantly associated with pathologic stage (odds ratio estimate was 1.075, 95% CI: 1.053-1.098). CONCLUSIONS: Quantitative histology, especially GPC, seems to be helpful for pre-operative staging of PCa in patients with T1c-T2 clinical stage and PSA < 10 ng/ml. PMID- 15780571 TI - Laparoscopic microwave tissue ablation of hepatic metastasis from a parathyroid carcinoma. PMID- 15780572 TI - Re: Selective lymphadenectomy in sentinel node-positive patients may increase the risk of local/in-transit recurrence in malignant melanoma, Thomas and Clark. PMID- 15780573 TI - Minimizing the release of proinflammatory and toxic bacterial products within the host: a promising approach to improve outcome in life-threatening infections. AB - Various bacterial components (e.g., endotoxin, teichoic and lipoteichoic acids, peptidoglycans, DNA) induce or enhance inflammation by stimulating the innate immune system and/or are directly toxic in eukariotic cells (e.g., hemolysins). When antibiotics which inhibit bacterial protein synthesis kill bacteria, smaller quantities of proinflammatory or toxic compounds are released in vitro and in vivo than during killing of bacteria by beta-lactams and other cell-wall active drugs. In general, high antibiotic concentrations liberate lower quantities of bacterial proinflammatory or toxic compounds than concentrations close to the minimum inhibitory concentration. In animal models of Escherichia coli Pseudomonas aeruginosa and Staphylococcus aureus peritonitis/sepsis and of Streptococcus pneumoniae meningitis, a lower release of proinflammatory bacterial compounds was associated with a reduced mortality or neuronal injury. Pre treatment with a bacterial protein synthesis inhibitor reduced the strong release of bacterial products usually observed during treatment with a beta-lactam antibiotic. Data available strongly encourage clinical trials comparing antibiotic regimens with different release of proinflammatory/toxic bacterial products. The benefit of the approach to reduce the liberation of bacterial products should be greatest in patients with a high bacterial load. PMID- 15780574 TI - High relative content of lysophospholipids of Helicobacter pylori mediates increased risk for ulcer disease. AB - Helicobacter pylori phospholipase A (OMPLA) degrades bacterial membrane phospholipids to lysophospholipids. High levels of lysophospholipids are associated with higher hemolytic activity, increased release of urease and vacA and better adherence to epithelial cells in vitro. The phospholipase A gene (pldA) displays phase variation due to a slippage in a homopolymeric tract. The aim of this study was to determine if the relative amount of lysophospholipids in the cell wall is associated with ulcer disease, and to further investigate the significance of pldA phase variation. H. pylori isolates of 40 patients were examined. The relative lysophospholipid content of each isolate was determined and the pldA gene was sequenced. The study indicated that H. pylori can regulate its OMPLA activity by phase variation in the pldA gene or by protein level regulation among phase variants in the pldA 'ON' status. We found a significant difference between the relative amount of lysophospholipids of the ulcer group and the non-ulcer group (p=0.022). When the lysophospholipid/phospholipid ratios were compared with outcome, the OR for ulcer disease was 9.0 (95% CI 1.6-49.4; p=0.014). Isolates with a high OMPLA activity are significantly associated with patients with ulcer disease. PMID- 15780575 TI - Strain-dependent disruption of blood-cerebrospinal fluid barrier by Streptoccocus suis in vitro. AB - Streptococcus suis capsular type 2 is an important agent of diseases including meningitis among pigs worldwide, and is also a zoonotic agent. The barrier function of the choroid plexus epithelium that constitutes the structural basis for the blood-cerebrospinal fluid (CSF) barrier has not been elucidated yet in bacterial meningitis. We investigated the influence of various S. suis isolates on the barrier function of cultured porcine choroid plexus epithelial cells with respect to the transepithelial resistance and paracellular [(3)H]-mannitol flux. Preferentially apical application of S. suis isolates significantly decreased transepithelial resistance and significantly increased paracellular [(3)H] mannitol flux in a time-, dose- and strain-dependent manner. Viable S. suis isolates caused cytotoxicity determined by lactate dehydrogenase assay and electron microscopy, whereas S. suis sonicates and UV-inactivated S. suis did not cause cytotoxicity. The observed effects on porcine choroid plexus epithelial cells barrier function could not exclusively be ascribed to known virulence factors of S. suis such as suilysin. In conclusion, S. suis isolates induce loss of blood-cerebrospinal fluid barrier function in an in vitro model. Thus, S. suis may facilitate trafficking of bacteria and leucocytes across the blood cerebrospinal fluid barrier. The underlying mechanisms for the barrier breakdown have yet to be determined. PMID- 15780576 TI - Effect of a booster dose of serogroup B meningococcal vaccine on antibody response to Neisseria meningitidis in mice vaccinated with different immunization schedules. AB - The generation and maintenance of memory antibody response by different primary immunization schedules with the Cuban-produced outer membrane protein based vaccine was investigated in a murine model. We analyzed the duration of the antibody response (IgG-ELISA and bactericidal titer) and the effect of a booster dose on the antibody response. The IgG avidity index was determined in an attempt to find a marker for memory development. This study also included an analysis of IgG subclasses induced by primary and booster immunization. The specificity of bactericidal antibodies was investigated using local strains of the same serotype/serosubtype (4,7:P1.19,15) as the vaccine strain and mutant strains lacking major outer membrane proteins. A significant recall response was induced by a booster dose given 7 months after a primary series of 2, 3 or 4 doses of vaccine. The primary antibody response showed a positive dose-effect. In contrast, a negative dose-effect was found on the booster bactericidal antibody response. There was a significant increase in IgG1 levels after the fourth and booster doses. Three doses of vaccine were required to induce a significant increase in IgG avidity. Two injections of vaccine induced a significant antibody response to PorA protein, while 4 injections induced a larger range of specificities. PMID- 15780577 TI - Altered T helper 1 reaction but not increase of virus load in patients with dengue hemorrhagic fever. AB - To investigate whether dengue-2 patients with and without dengue hemorrhagic fever had different virus load, immune mediators, or T helper (Th) reaction, we simultaneously measured virus load, immune mediators and the Th1/Th2 transcription factors T-bet/GATA-3 mRNA expression in a large outbreak of dengue 2 infections in Southern Taiwan. Results showed that virus load was not significantly different between patients with and without dengue hemorrhagic fever. Patients with dengue fever had higher IFN-gamma levels, but patients with dengue hemorrhagic fever had significantly higher IL-10 levels. Further studies showed that patients with dengue hemorrhagic fever had a significantly lower T bet than those with dengue fever, but GATA-3 mRNA expression in peripheral blood leukocytes was not significant difference between both groups. In conclusion, altered Th1 reaction as reflected by lower T-bet mRNA expression associated with higher IL-10 levels might be involved in the pathogenesis of dengue hemorrhagic fever. PMID- 15780579 TI - Mucosal immunisation with novel Streptococcus pneumoniae protein antigens enhances bacterial clearance in an acute mouse lung infection model. AB - Streptococcus pneumoniae contains many proteins that have not been evaluated as potential protective vaccine antigens. In this study we isolated proteins from a serotype 3 strain of S. pneumoniae for use in mouse immunisation studies. Separation of the protein mix was achieved by SDS-PAGE electrophoresis followed by electro-elution to isolate individual proteins. This procedure successfully separated 21 fractions from which six proteins were selected based on purity and quantity and were initially denoted by their molecular masses: 14-, 34-, 38-, 48 , 57- and 75-kDa. The immunogenicity of these proteins was investigated in a mucosal immunisation model in mice involving a primary inoculation to the intestinal Peyer's patches followed by an intra-tracheal boost two weeks later. The immune response was assessed by enhancement of pulmonary clearance of infection, recruitment of phagocytes to the lungs and induction of an antibody response. Two of the proteins, the 14-kDa identified as a L7/L12 ribosomal protein, and the 34-kDa identified as glyceraldehyde-3-phosphate dehydrogenase resulted in up to 99% and 94%, respectively, enhanced clearance of infection within 5 h following pulmonary challenge with S. pneumoniae. This study has shown that novel pneumococcal proteins have the potential to be vaccine candidates to enhance clearance of an acute mucosal S. pneumoniae infection. PMID- 15780578 TI - Mechanisms of Porphyromonas gingivalis-induced monocyte chemoattractant protein-1 expression in endothelial cells. AB - Monocyte chemoattractant protein-1 (MCP-1) is expressed in vascular endothelial cells of inflamed gingival tissues and plays an important role in periodontal pathogenesis. Endothelial cells produce high levels of MCP-1 in response to Porphyromonas gingivalis, an important periodontal pathogen. The present study investigated the mechanisms involved in MCP-1 production by human umbilical vein endothelial cells (HUVEC) following infection with P. gingivalis. In contrast to P. gingivalis, Bacteroides forsythus only weakly stimulated MCP-1 production while Treponema denticola could not induce MCP-1 in HUVEC. The MCP-1 production was independent of endogenous interleukin (IL)-1alpha as IL-1 receptor antagonist treatment did not reduce MCP-1 production by P. gingivalis. Meanwhile, antioxidant treatment and inhibition of NAD(P)H oxidase significantly reduced MCP 1 production. Pharmacological inhibition of p38 mitogen-associated protein (MAP) kinase, c-Jun N-terminal kinase (JNK), nuclear factor-kappaB (NF-kappaB) or activator protein-1 (AP-1) also substantially attenuated P. gingivalis-induced MCP-1 expression by HUVEC. Indeed, activation of NF-kappaB and AP-1 was observed in P. gingivalis-infected HUVEC. These results suggest that MCP-1 expression is upregulated in P. gingivalis-infected endothelial cells via reactive oxygen species, p38 MAP kinase, JNK, NF-kappaB, and AP-1. PMID- 15780580 TI - Expression of a peroxiredoxin-glutaredoxin by Haemophilus influenzae in biofilms and during human respiratory tract infection. AB - Evidence is mounting that nontypeable Haemophilus influenzae grows as a biofilm in the middle ear of children with otitis media and the airways of adults with chronic obstructive pulmonary disease. To begin to assess antigens expressed by H. influenzae in biofilms, cell envelopes of bacteria grown as a biofilm were compared to those grown planktonically. A approximately 30kDa peroxiredoxin glutaredoxin was present in greater abundance during growth in biofilms. Mutants deficient in expression of peroxiredoxin-glutaredoxin were constructed by homologous recombination in four clinical isolates. The mutants showed a 25-50% reduction in biofilm formation compared to the corresponding parent strains. To study in vivo expression of peroxiredoxin-glutaredoxin during human respiratory tract infection, paired pre- and post-exacerbation serum from adults with chronic obstructive pulmonary disease and H. influenzae in sputum were assayed using an enzyme-linked immunosorbent assay and purified recombinant peroxiredoxin glutaredoxin. Eight from 18 (44.4%) paired serum samples showed a significant increase in antibody to peroxiredoxin-glutaredoxin from pre- to post-infection. These results indicate that (1) peroxiredoxin-glutaredoxin is present in greater abundance in H. influenzae biofilms compared to planktonically grown bacteria; (2) peroxiredoxin-glutaredoxin is involved in biofilm formation by H. influenzae and the degree of involvement varies among strains; and (3) peroxiredoxin glutaredoxin is expressed by H. influenzae during infection of the human respiratory tract and is recognized by the human immune system. PMID- 15780581 TI - Ribotyping of Burkholderia mallei isolates. AB - In this study, the subspecies differentiation of 25 isolates of Burkholderia mallei was attempted based on their ribotype polymorphisms. The isolates were from human and equine infections that occurred at various times around the world. DNA samples from each isolate were digested separately with PstI and EcoRI enzymes and probed with an Escherichia coli-derived 18-mer rDNA sequence to identify diagnostic fragments. Seventeen distinct ribotypes were identified from the combined data obtained with the two restriction enzymes. The results demonstrate the general utility of ribotyping for the subspecies identification of B. mallei isolates. PMID- 15780584 TI - Compartmentalization of growth factor receptor signalling. AB - Spatial and temporal separation of signal transduction pathways often determines the specificity in cellular responses. Recent advances have improved our understanding of how growth factor signalling is influenced by the formation of molecular complexes (signalosomes) in distinct cellular compartments. There has also been new insight into the mechanisms that determine the signalling competence of these complexes and their role in receptor endocytosis, retrograde trafficking in neurons and restricted protein biosynthesis, and many examples have been found where signalosome deregulation leads to disease. PMID- 15780582 TI - Role of anti-beta-glucan antibody in host defense against fungi. AB - We have recently detected an anti-beta-glucan antibody in normal human and normal mouse sera. The anti-beta-glucan antibody showed reactivity to pathogenic fungal Aspergillus and Candida cell wall glucan. Anti-beta-glucan antibody could bind whole Candida cells. It also enhanced the candidacidal activity of macrophages in vitro. The anti-beta-glucan antibody titer of DBA/2 mice intravenously administered either Candida or Aspergillus solubilized cell wall beta-glucan decreased remarkably dependent on dose. Moreover, in deep mycosis patients, the anti-beta-glucan antibody titer decreased, and this change correlated with clinical symptoms and other parameters such as C-reactive protein. It was suggested that the anti-beta-glucan antibody formed an antigen-antibody complex and participated in the immune response as a molecule recognizing pathogenic fungi. PMID- 15780585 TI - Axonal growth: where neurotrophins meet Wnts. AB - Axonal guidance is influenced by many cues, including polypeptide trophic factors, cytokines, diffusible attractants and repellents and changes in calcium. How these signals are conveyed and integrated is not well defined. Recent data suggest that molecules of the canonical Wnt signaling pathway may have direct actions on axonal growth through neurotrophin signaling. This surprising mechanism is supported by local inactivation of glycogen synthase kinase 3beta (GSK-3beta) by nerve growth factor through the integrin-linked kinase. Inhibition of GSK-3beta provides a positive regulatory signal for the cytoskeleton re arrangement involved in axon extension. Moreover, microtubule stabilization is stimulated by adenomatous polyposis coli protein, a downstream target of GSK3, in response to neurotrophins. Therefore, components of the Wnt signaling pathway are downstream of trophic factors, providing new insights into cytoskeletal regulatory events during axonal growth. PMID- 15780586 TI - Peptide signalling in plant development and self/non-self perception. AB - Plant genomes contain very large families of genes encoding receptor-like kinases (RLKs). In recent years, several of these RLKs have been shown by biochemical and mutational analysis to represent receptors for peptides, and the emerging picture shows that peptide signalling in development and self/non-self perception is based on a similar repertoire of receptors and signalling cascades. The need to recognize multiple peptide signals in self/non-self recognition may have led to the surprising radiation and diversification of RLKs in the plant kingdom. PMID- 15780587 TI - Linking Rap to cell adhesion. AB - The small GTPase Rap1 is involved in several aspects of cell adhesion, including integrin-mediated cell adhesion and cadherin-mediated cell junction formation. Recently, several effector proteins for Rap1 have been identified providing a clear link between Rap1 and actin dynamics. Furthermore, evidence is accumulating that Rap1 functions in the spatial and temporal control of cell polarity. PMID- 15780588 TI - Arrestin times for compartmentalised cAMP signalling and phosphodiesterase-4 enzymes. AB - Various methods reveal that cyclic AMP (cAMP) signalling in cells is compartmentalised. These methods use FRET probes based upon either protein kinase A (PKA) or EPAC, cAMP-gated ion channels, or the selective activation of AKAP anchored PKA isoforms. The basis of compartmentalisation involves point sources of cAMP generation within sub-domains of the plasma membrane coupled to degradation by spatially segregated, anchored forms of cAMP phosphodiesterases. cAMP-specific phosphodiesterase-4 (PDE4) isoforms play a central role in determining compartmentalisation, as exemplified in cardiac myocytes and T cells. The PKA phosphorylation status of the beta2-adrenoreceptor, and hence its ability to switch its signalling from G(s) to G(i) and thus to activate ERK, is regulated dynamically by the agonist-stimulated recruitment of PDE4 to the receptor in complex with beta-arrestin. The co-receptor CD28 enhances signalling through the T-cell receptor by recruiting a PDE4/beta-arrestin complex, which then attenuates PKA phosphorylation of Csk. PMID- 15780589 TI - A plethora of interacting organellar Ca2+ stores. AB - The endoplasmic reticulum is not the only major agonist-releasable Ca2+ store within cells; it is now clear that virtually all organelles so far studied have the ability to act as mobilizable Ca2+ stores. From recent findings with regard to Ca2+ transportation and Ca2+ homeostasis within a variety of cell organelles such as the mitochondria, nucleus, Golgi and lysosomes, it emerges that many of these organellar Ca2+ stores appear to interact with each other, adding a further level of complexity to Ca2+ signalling events. PMID- 15780590 TI - Phosphoinositide 3-kinase in disease: timing, location, and scaffolding. AB - When PI3Ks are deregulated by aberrant surface receptors or modulators, accumulation of PtdIns(3,4,5)P3 leads to increased cell growth, proliferation and contact-independent survival. The PI3K/PKB/TOR axis controls protein synthesis and growth, while PtdIns(3,4,5)P3-mediated activation of Rho GTPases directs cell motility. PI3K activity has been linked to the formation of tumors, metastasis, chronic inflammation, allergy and cardiovascular disease. Although increased PtdIns(3,4,5)P3 is a well-established cause of disease, it is seldom known which PI3K isoform is implied. Recent work has demonstrated that PI3Kgamma contributes to the control of cAMP levels in the cardiac system, where the protein acts as a scaffold, but not as a lipid kinase. PMID- 15780591 TI - Recent advances in the protein kinase B signaling pathway. AB - The phosphoinositide 3' kinase signaling pathway is activated in response to a plethora of growth factors and cytokines, and initiates a cascade of signaling events primarily via the induction of specific protein-serine/threonine kinases. Interest in the pathway has been driven by its frequent aberrant activation in disease and its impact on cell fate decisions owing to roles in survival signaling and metabolic control. There have been recent advances in our understanding of the primary components of this pathway, namely phosphoinositide dependent kinase-1, protein kinase B and glycogen synthase kinase-3, including insights into their mechanisms of regulation, substrate proteins and cellular functions. PMID- 15780592 TI - The expanding TOR signaling network. AB - Cell growth (increase in cell mass or size) is tightly coupled to nutrient availability, growth factors and the energy status of the cell. The target of rapamycin (TOR) integrates all three inputs to control cell growth. The discovery of upstream regulators of TOR (AMPK, the TSC1-TSC2 complex and Rheb) has provided new insights into the mechanism by which TOR integrates its various inputs. A recent finding in flies reveals that TOR controls not only growth of the cell in which it resides (cell-autonomous growth) but also the growth of distant cells, thereby determining organ and organism size in addition to the size of isolated cells. In yeast and mammals, the identification of two structurally and functionally distinct multiprotein TOR complexes (TORC1 and TORC2) has provided a molecular basis for the complexity of TOR signaling. Furthermore, TOR has emerged as a regulator of growth-related processes such as development, aging and the response to hypoxia. Thus, TOR is part of an intra- and inter-cellular signaling network with a remarkably broad role in eukaryotic biology. PMID- 15780593 TI - New roles for the LKB1-->AMPK pathway. AB - The AMP-activated protein kinase (AMPK) is a sensor of cellular energy that is conserved throughout eukaryotes. It is activated by rising AMP, signifying falling energy status caused by starvation for a carbon source or other stress. Binding of AMP to the regulatory gamma subunit triggers phosphorylation of the catalytic alpha subunit by the upstream kinase LKB1, and the activated kinase switches on ATP-generating catabolic pathways while switching off ATP-requiring processes. AMPK inhibits the TOR (target of rapamycin) pathway by phosphorylating TSC2, thus inhibiting cell growth during times of stress. AMPK is also a target for adipokines that regulate energy balance at the whole-body level. PMID- 15780594 TI - Cyclooxygenases, peroxide tone and the allure of fish oil. AB - Skepticism about the health benefits of fish oil is largely the result of our incomplete understanding of the biochemistry of omega3 essential fatty acids. Recent work has confirmed the roles of omega3 fatty acids in gene transcription and signal transduction, and has given insight into the effects of eicosapentaenoic acid (EPA) and the EPA/arachidonic acid (AA) ratio on prostanoid (PG) metabolism and function. One pronounced effect of fish-oil-induced increases in EPA/AA ratios is decreased PG formation from AA via cyclooxygenase-1, because EPA inhibits this isoform. In addition, cells lacking endogenous alkyl-peroxide generating systems and thus having a low 'peroxide tone' cannot oxygenate EPA via cyclooxygenase-1. Platelets, however, which are equipped with a lipoxygenase that can produce an abundance of hydroperoxide from AA, can form small amounts of thromboxane A3 from EPA via cyclooxygenase-1. A second major consequence of elevated EPA/AA ratios is significantly increased production of 3-series PGs, including PGE3, via cyclooxygenase-2. There are four PGE receptor subtypes and at least one of these types--not yet identified--has a significantly different response to PGE3 than to PGE2; this difference may underlie the ability of omega3 fatty acids to mitigate inflammation and tumorigenesis. PMID- 15780595 TI - Intracellular messenger function of hydrogen peroxide and its regulation by peroxiredoxins. AB - Hydrogen peroxide (H2O2) accumulates transiently in various cell types stimulated with peptide growth factors and participates in receptor signaling by oxidizing the essential cysteine residues of protein tyrosine phosphatases and the lipid phosphatase PTEN. The reversible inactivation of these phosphatases by H2O2 is likely required to prevent futile cycles of phosphorylation-dephosphorylation of proteins and phosphoinositides. The accumulation of H2O2 is possible even in the presence of large amounts of the antioxidant enzymes peroxiredoxin I and II in the cytosol, probably because of a built-in mechanism of peroxiredoxin inactivation that is mediated by H2O2 and reversed by an ATP-dependent reduction reaction catalyzed by sulfiredoxin. PMID- 15780596 TI - Fatty acylation and prenylation of proteins: what's hot in fat. AB - Post-translational modification by covalent attachment of lipid groups helps proteins to associate with membranes, both intra- and extracellularly. The enzymology of protein S-acylation with fatty acids has been a stumbling block, but three pathways for this modification have now been identified in eukaryotes. It is not yet clear whether this reaction is enzymatic or facilitated by a chaperone-like mechanism. Work with Ras proteins has shown that an S acylation/deacylation cycle, in cooperation with prenylation and carboxyl methylation, may regulate their cycling between intracellular membrane compartments and subdomains, hence controlling their signalling activity. The two types of prenyl group, geranylgeranyl and farnesyl, themselves have surprisingly specific targeting roles for Ras superfamily members. PMID- 15780597 TI - Protein serine/threonine phosphatases: life, death, and sleeping. AB - Protein serine/threonine phosphatases control key biological pathways including early embryonic development, cell proliferation, cell death, circadian rhythm and cancer. Recent studies have provided important insights into how several of the many phosphatase regulators, through their interaction with a conserved phosphatase catalytic subunit, control the activity of critical substrates in these diverse pathways. Recent co-crystal structures provided a major insight into how the diverse protein serine/threonine regulators rein in the otherwise promiscuous catalytic subunits. PMID- 15780598 TI - Cytoplasmic protein tyrosine phosphatases, regulation and function: the roles of PTP1B and TC-PTP. AB - PTP1B and TC-PTP are closely related protein tyrosine phosphatases, sharing 74% homology in their catalytic domain. However, their cellular localization, function, and regulation are found to be different. Their substrate specificity has implicated these enzymes in various signaling pathways, regulating metabolism, proliferation and cytokine signaling. For instance, PTP1B has been shown to regulate the activation of cytokine receptors through the dephosphorylation of specific members of the JAK family, namely JAK2 and TYK2, whereas TC-PTP is involved in the modulation of cytokine signaling via JAK1 and JAK3 molecules. Gene-targeting approaches will help us to unravel the physiological functions of these enzymes. PMID- 15780599 TI - Molecular chaperones as regulatory elements of cellular networks. AB - Molecular chaperones help hundreds of signaling molecules to keep their activation-competent state, and regulate various signaling processes ranging from signaling at the plasma membrane to transcription. Besides these specific regulatory roles, recent studies have revealed that chaperones act as genetic buffers stabilizing the phenotypes of various cells and organisms. This may be related to their low affinity for the proteins they interact with, which means that they represent weak links in protein networks. Chaperones may uncouple protein, signaling, membrane, organelle and transcriptional networks during stress, which gives the cell additional protection. The same networks are preferentially remodeled in various diseases and aging, which may help us to design novel therapeutic and anti-aging strategies. PMID- 15780600 TI - Kinase signaling cascades that modulate peroxisome proliferator-activated receptors. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors involved in lipid and glucose homeostasis, inflammation and wound healing. In addition to ligand binding, phosphorylation can also regulate PPARs; the biological effects of phosphorylation depend on the stimulus, the kinase, the PPAR isotype, the residue modified, the cell type and the promoter investigated. The study of this dual regulation mode, which allows PPARs to integrate signals conveyed by lipophilic ligands with those coming from the plasma membrane, may ultimately offer new therapeutic strategies. PMID- 15780601 TI - Cellular oscillators: rhythmic gene expression and metabolism. AB - Many biological processes are driven by biological clocks that, depending on the frequency they generate, are classified into ultradian, circadian and infradian oscillators. In virtually all light-sensitive organisms from cyanobacteria to humans, a circadian timing system adapts cyclic physiology to geophysical time. Recent evidence suggests that even in mammals circadian oscillators function in a cell-autonomous manner. In yeast, an ultradian oscillator regulates cyclic respiratory activity and global gene expression. Circadian oscillators and the ultradian yeast respiratory clock share at least four properties: they follow limit-cycle kinetics, interweave with cellular metabolism, are temperature compensated and influence the cell division clock. PMID- 15780602 TI - Methylation of histones: playing memory with DNA. AB - Nucleosomal histones can be methylated in vivo at multiple residues and defined methylation patterns are related to distinct functional readouts of chromosomal DNA. Histone methylation has emerged as an important post-translational modification involved in transcriptional regulation and genome integrity. Recent progress in determining the cis and trans determinants of this process revealed multiple roles for histone methylation in epigenetic memory of active and silent states. The analysis of imprinted, X-linked and heterochromatic sequences disclosed mechanistic similarities for heritable transcriptional repression, pointing to a common mode of action. Moreover, the view of histone methylation as a stable modification has recently been challenged by studies revealing a number of pathways that are capable of removing histone methylation. Thus, in addition to having great in vivo complexity, this modification appears more dynamic then was previously thought. PMID- 15780603 TI - Novel 2-aminothiazonaphthalimides as visible light activatable photonucleases: effects of intercalation, heterocyclic-fused area and side chains. AB - A new family of 2-aminothiazonaphthalimides with different side chains as novel intercalative and visible light activatable photonucleases, was designed, synthesized and quantitatively evaluated. The order of their photocleaving abilities was parallel to that of their intercalative properties. The compound with linear heterocyclic-fused chromophore could intercalate into and photocleave DNA more efficiently than the one with angular heterocyclic-fused chromophore. B(2), the most efficient compound, caused obvious DNA damage at 1 microM. Mechanism experiment showed that superoxide anion was involved. PMID- 15780604 TI - 18F-labeled sufentanil for PET-imaging of mu-opioid receptors. AB - The synthesis of an (18)F-labeled sufentanil analogue with apparent high mu opioid receptor selectivity is reported. Intravenous injection of N-[4 (methoxymethyl)-1-[2-(2-thienyl)ethyl]-4-piperidinyl]-N-phenyl-2-(+/-) [(18)F]fluoropropan-amide in mice resulted in high brain uptake and a regional brain activity distribution corresponding to the mu-opioid receptor expression pattern. The developed ligand is a promising tracer for extended protocols in mu opioid receptor mapping and quantitation with positron emission tomography. PMID- 15780605 TI - QSAR-by-NMR: quantitative insights into structural determinants for binding affinity by analysis of 1H/15N chemical shift differences in MMP-3 ligands. AB - A novel strategy is applied to obtain quantitative insights on factors influencing biological affinity in protein-ligand complexes. This approach is based on the detection of ligand binding by (15)N and (1)H amide chemical shift differences in two-dimensional (15)N-heteronuclear single-quantum correlation spectra. Essential structural features linked to affinity can be extracted using statistical analysis of (15)N and (1)H amide chemical shift differences in congeneric series relative to uncomplexed protein spectra, as demonstrated for 20 MMP-3 inhibitors in complex with human matrix metalloproteinase stromelysin (MMP 3). The statistical analysis using PLS led to a significant model, while its chemical interpretation, highlighting the importance of particular residues for affinity, are in agreement to an X-ray structure of one key compound in the homologue MMP-8 binding site. PMID- 15780606 TI - Quinolinium salt as a potent inhibitor of lymphocyte apoptosis. AB - The synthesis of several quinolinium salts and related compounds and their ability to inhibit glucocorticoid-induced apoptosis in murine thymocytes are described. Interestingly, 1-[2-methoxyimino-2-(4-pyrrolidin-1-yl phenyl)ethyl]quinolinium bromide (11) showed a potent protective effect with an EC(50) of 0.013 microM, which was at least 300-fold more potent than the reference compound pifithrin-alpha. PMID- 15780607 TI - Structure-activity relationships of the oxindole growth hormone secretagogues. AB - A series of substituted oxindole derivatives of SM-130686 was synthesized and evaluated as ghrelin receptor agonists. Modification of the substituents on the C3-aromatic part of the oxindole led to compounds with subnanomolar binding affinities. Compound 4i (IC(50)=0.02 nM) was orally active at low doses and showed in vivo activity when orally administered, 2 mg/kg twice a day for 4 days, as evidenced by significant body weight gain. PMID- 15780608 TI - Design, synthesis, biological evaluation and molecular docking of curcumin analogues as antioxidant, cyclooxygenase inhibitory and anti-inflammatory agents. AB - Curcuminoids were isolated from Curcuma longa and their pyrazole and isoxazole analogues were synthesized and evaluated for antioxidant, COX-1/COX-2 inhibitory and anti-inflammatory activities. The designed analogues significantly enhance COX-2/COX-1 selectivity and possess significant anti-inflammatory activity in carrageenan induced rat paw edema assay. Pyrazole, isoxazole analogues of curcumin (4 and 7) exhibited higher antioxidant activity than trolox. Molecular docking study revealed the binding orientations of curcumin analogues in the active sites of COX and thereby helps to design novel potent inhibitors. PMID- 15780609 TI - Tetrahydroisoquinoline 1-carboxamides as growth hormone secretagogues. AB - Several novel series of tetrahydroisoquinoline 1-carboxamides were prepared and shown to be potent growth hormone (GH) secretagogues. Among them, carbamate 12a E2 displays excellent in vivo activity by increasing plasma GH 10-fold in an anesthetized IV rat model. PMID- 15780610 TI - Novel fluoroquinolones: design, synthesis, and in vivo activity in mice against Mycobacterium tuberculosis H37Rv. AB - Novel 6,8-difluoro-1-alkyl-5-amino-1,4-dihydro-4-oxo-7-{4-substituted piperazin-1 yl}-quinoline-3-carboxylic acids, with the substituents at 4th position of piperazine being -[2(pyridine-4-carbonyl) hydrazono]propyl and -2 [(pyrazine-2 carbonyl) amino] ethyl, were synthesized and evaluated in vivo against Mycobacterium tuberculosis H(37)Rv in Swiss albino mice. Test compounds exhibited activity comparable to that of sparfloxacin (survival rate, reduction of splenomegaly and reduced tubercular lesions) at a dose of 200 mg/kg. PMID- 15780611 TI - Potent pyrimidinetrione-based inhibitors of MMP-13 with enhanced selectivity over MMP-14. AB - Through the use of computational modeling, a series of pyrimidinetrione-based inhibitors of MMP-13 was designed based on a lead inhibitor identified through file screening. Incorporation of a biaryl ether moiety at the C-5 position of the pyrimidinetrione ring resulted in a dramatic enhancement of MMP-13 potency. Protein crystallography revealed that this moiety binds in the S(1)(') pocket of the enzyme. Optimization of the C-4 substituent of the terminal aromatic ring led to incorporation of selectivity versus MMP-14 (MT-1 MMP). Structure activity relationships of the biaryl ether substituent are presented as is pharmacokinetic data for a compound that meets our in vitro potency and selectivity goals. PMID- 15780612 TI - Enhancement in antimicrobial activity of 2-(phenyl)-3-(2-butyl-4-chloro-1H imidazolyl)-5-butylate isoxazolidine. AB - The trans rich isomer, 2-(phenyl)-3-(2-butyl-4-chloro-1H-imidazolyl)-5-butylate isoxazolidine A (>96% ee) was synthesized by the condensation of E isomer rich nitrone 4 (>98% ee) with butyl acrylate in an inert solvent. Obtained isoxazolidine was screened for its antifungal activity against Aspergillus niger, Cephalosporium acremonium, Fusarium moniliforme by using Nystatin as positive control. It was also tested for its antibacterial activity against Bacillus subtilis, Escherichia coli, and Staphylococcus aureus by using Streptomycin as positive control. Enhanced antifungal activity was observed in isoxazolidine of >96% ee compared to the isoxazolidine of >69% ee (B), and enhancement was not observed in antibacterial activity. PMID- 15780613 TI - Novel and potent cyclic cyanamide-based cathepsin K inhibitors. AB - Starting from a PDE IV inhibitor hit derived from high throughput screening of the compound collection, a key pyrrolidine cyanamide pharmacophore was identified. Modifications of the pyrrolidine ring produced enhancements in cathepsin K inhibition. An X-ray co-crystal structure of a cyanamide with cathepsin K confirmed the mode of inhibition. PMID- 15780614 TI - Asymmetric synthesis of novel tetrahydroquinoline derivatives with a sugar building block and their bioactivities. AB - Some novel spiro-tetrahydroquinolines were stereoselectively synthesized by using keto-sugar derived from sucrose as a building block in one pot under mild conditions. The in vitro immunobiological activity and cytotoxicity of these novel tetrahydroquinolines were investigated. The results implied that these spiro-compounds have obvious bioactivity and may be structurally modified to improve bioactivity further. PMID- 15780615 TI - Structure-activity relationship studies on tetralin carboxamide growth hormone secretagogue receptor antagonists. AB - The structure-activity relationship studies on a series of tetralin carboxamide growth hormone secretagogue receptor (GHS-R) antagonists are discussed. It was found that certain 2-alkoxycarbonylamino substituted tetralin carboxamides are potent, selective, and orally bioavailable GHS-R antagonists. PMID- 15780616 TI - Fused pyrimidine based inhibitors of phosphodiesterase 7 (PDE7): synthesis and initial structure-activity relationships. AB - A series of fused pyrimidine based inhibitors of PDE7 have been derived from an earlier screening lead 1. The synthesis, structure-activity relationships (SAR) and selectivity against several other PDE family members are described. PMID- 15780617 TI - Novel heterocyclic thyromimetics. AB - Novel heterocycle-fused thyromimetics are presented carrying indoles or indazoles instead of the phenolic group in T3. Potent agonists were identified in both series. SAR trends are examined and found to be mostly consistent with previously published thyromimetics. Moderate THRbeta selectivity (approx. 10-fold) was observed in the indole series using isoform-selective transient THR transfection assays. PMID- 15780618 TI - 1,3-Dioxo-4-methyl-2,3-dihydro-1H-pyrrolo[3,4-c]quinolines as potent caspase-3 inhibitors. AB - Synthesis, biological evaluation and structure-activity relationships for a series of novel nonpeptide small molecule inhibitors of caspase-3 are described. Among the studied compounds, 8-sulfamide derivatives of 1,3-dioxo-4-methyl-2,3 dihydro-1H-pyrrolo[3,4-c]quinolines have been identified as potent inhibitors of caspases-3. The most active compound within this series (8f) inhibited caspase-3 with IC(50)=4 nM. PMID- 15780619 TI - Structure-activity relationship of the novel bivalent and C-terminal modified analogues of endomorphin-2. AB - Endomorphin-2 (Tyr-Pro-Phe-Phe-NH(2)) is a putative endogenous mu-opioid receptor ligand. To develop potent analgesics with less side effects related to it, we used the methods of dimerization and C-terminal modification. Through dimerization we got the 'balanced agonists' with potent analgesic activity and we have developed the structure-activity relationship between the selectivity and the distance of the two tyrosine pharmacophores. Modification at the C-terminal increased the selectivity of endomorphin-2 to mu-opioid receptor with binding affinity conserved. PMID- 15780620 TI - Fluorescent metal ion indicators based on benzoannelated crown systems: a green fluorescent indicator for intracellular sodium ions. AB - The synthesis and metal binding properties of cation-sensitive fluorescent indicators intended for biological applications are described. The increase of the crown ether ring size enhances the affinity for larger cations, but weakens the fluorescent response and selectivity. A compound having a 15-crown-5 chelator directly attached to a 2,7-difluoroxanthenone fluorophore loads into live cells and responds to sodium ion concentration changes with large fluorescence increases in the visible wavelength range. PMID- 15780621 TI - Structure-based design, synthesis, and biological evaluation of novel 1,4 diazepines as HDM2 antagonists. AB - Crystallographic analysis of ligands bound to HDM2 suggested that 7-substituted 1,4-diazepine-2,5-diones could mimic the alpha-helix of p53 peptide and may represent a promising scaffold to develop HDM2-p53 antagonists. To verify this hypothesis, we synthesized and biologically evaluated 5-[(3S)-3-(4-chlorophenyl) 4-[(R)-1-(4-chlorophenyl)ethyl]-2,5-dioxo-7-phenyl-1,4-diazepin-1-yl]valeric acid (10) and 5-[(3S)-7-(2-bromophenyl)-3-(4-chlorophenyl)-4-[(R)-1-(4 chlorophenyl)ethyl]-2,5-dioxo-1,4-diazepin-1-yl]valeric acid (11). Preliminary in vitro testing shows that 10 and 11 substantially antagonize the binding between HDM2 and p53 with an IC(50) of 13 and 3.6 microM, respectively, validating the modeling predictions. Taken together with the high cell permeability of diazepine 11 determined in CACO-2 cells, these results suggest that 1,4-diazepine-2,5 diones may be useful in the treatment of certain cancers. PMID- 15780622 TI - Synthesis and anticonvulsant activity of new 2-substituted-5-(2-benzyloxyphenyl) 1,3,4-oxadiazoles. AB - A series of new 2-substituted-5-(2-benzyloxyphenyl)-1,3,4-oxadiazoles have been synthesized and evaluated as anticonvulsant agents. Compound 4b shows considerable anticonvulsant activity both in PTZ and MES models. It seems this effect is mediated through benzodiazepine receptors mechanism. PMID- 15780623 TI - Structure-based design of derivatives of tyropeptin A as the potent and selective inhibitors of mammalian 20S proteasome. AB - Tyropeptin A, a new potent proteasome inhibitor, was produced by Kitasatospora sp. MK993-dF2. To enhance the inhibitory potency of tyropeptin A, we constructed the structural model of tyropeptin A bound to the site responsible for the chymotrypsin-like activity of mammalian 20S proteasome. Based on these modeling experiments, we designed and synthesized several derivatives of tyropeptin A. Among them, the most potent compound, TP-104, exhibited a 20-fold enhancement in its inhibitory potency compared to tyropeptin A. Additionally, TP-110 specifically inhibited the chymotrypsin-like activity, but did not inhibit the PGPH and the trypsin-like activities. PMID- 15780624 TI - Synthesis of 15R-PGD2: a potential DP2 receptor agonist. AB - The first total synthesis of 15R-PGD(2)3 was accomplished. The approach used in this report is also an efficient method to produce 15R-PGE(2). 15R-PGD(2), a potential DP(2) receptor agonist, could be an important novel tool for defining the role of this receptor in inflammatory diseases. PMID- 15780625 TI - Synthesis and pharmacological investigation of novel 1-substituted-4-phenyl-1,2,4 triazolo[4,3-a]quinazolin-5(4H)-ones as a new class of H1-antihistaminic agents. AB - A series of novel 1-substituted-4-phenyl-1,2,4-triazolo[4,3-a]quinazolin-5(4H) ones 7 were synthesized by the cyclization of 2-hydrazino-3-phenylquinazolin 4(3H)-one 6 with various one carbon donors. The starting material 2-hydrazino-3 phenylquinazolin-4(3H)-one 6, was synthesized from aniline 1 by a novel innovative route. When tested for their in vivo H(1)-antihistaminic activity on conscious guinea pigs all the test compounds protected the animals from histamine induced bronchospasm significantly, whereas the compound 1-methyl-4-phenyl-1,2,4 triazolo[4,3-a]quinazolin-5(4H)-one 7b (percentage protection 70.7%) was found to be equipotent with the reference standard chlorpheniramine maleate (percentage protection 71%). These compounds show negligible sedation ( approximately 5%) when compared to the reference standard (26%). Hence they could serve as prototype molecules for future development. PMID- 15780626 TI - Antimalarial activity of 2,4,6-trisubstituted pyrimidines. AB - A series of 2,4,6-trisubstituted pyrimidines (3a-o) was synthesized and evaluated for their in vitro antimalarial activity against P. falciparum. Out of the 15 compounds synthesized 11 compounds showed MIC in the range of 0.5-2 microg/mL. These compounds are in vitro several folds more active than pyrimethamine. PMID- 15780628 TI - Molecular-modeling based design, synthesis, and activity of substituted piperidines as gamma-secretase inhibitors. AB - Alzheimer's disease (AD) is a debilitating disease widely thought to be associated with the accumulation of beta amyloid (Abeta) in the brain. Inhibition of gamma-secretase, one of the enzymes responsible for Abeta production, may be a useful strategy for the treatment of AD. Described below is a series of gamma secretase inhibitors designed from a scaffold identified by a ROCS [J. Comput. Chem.1996, 17, 1653] search of the corporate database. PMID- 15780627 TI - Structure-activity relationship study of novel tissue transglutaminase inhibitors. AB - Thieno[2,3-d]pyrimidin-4-one acylhydrazide derivatives were discovered as moderately potent inhibitors of TGase 2 (tissue transglutaminase) utilizing a fluorescence-based assay that measured TGase 2 catalyzed incorporation of the dansylated Lys derivative alpha-N-Boc-Lys-CH(2)-CH(2)-dansyl into the protein substrate N,N-dimethylated-casein. A SAR study revealed that the acylhydrazide thioether side-chain and the thiophene ring were critical to inhibitory activity. PMID- 15780629 TI - Design, synthesis, and structure-activity relationships of tetrahydroquinoline based farnesyltransferase inhibitors. AB - Tetrahydroquinoline-based small molecule inhibitors of farnesyltransferase (FT) have been identified. Lead compounds were shown to have nanomolar to sub nanomolar activity in biochemical assays with excellent potency in a Ras-mutated cellular reversion assay. BMS-316810 (9e), a 0.7 nM FT inhibitor, was orally active in a nude mouse tumor allograft efficacy study. PMID- 15780630 TI - Novel cyclopentane dicarboxamide sodium channel blockers as a potential treatment for chronic pain. AB - A series of new voltage-gated sodium channel blockers were prepared based on the screening lead succinic diamide BPBTS. Replacement of the succinimide linker with the more rigid cyclic 1,2-trans-diamide linker was well tolerated. N-Methylation on the biphenylsulfonamide side of the amide moiety significantly reduced the clearance rate in rat pharmacokinetic studies. PMID- 15780631 TI - Carbonic anhydrase inhibitors. Inhibition of isozymes I, II, IV, V and IX with complex fluorides, chlorides and cyanides. AB - The inhibition of five human carbonic anhydrase (hCA, EC 4.2.1.1) isozymes, the cytosolic hCA I and II, the membrane-bound hCA IV, the mitochondrial hCA V and the tumour associated, transmembrane hCA IX, with complex anions incorporating fluoride, chloride and cyanide, as well as B(III), Si(IV), P(V), As(V), Al(III), Fe(II), Fe(III), Pd(II), Pt(II), Pt(IV), Cu(I), Ag(I), Au(I) and Nb(V) species has been investigated. Apparently, the most important factors influencing activity of these complexes are the nature of the central metal ion/element, and its charge. Geometry of these compounds appears to be less important, since both linear, tetrahedral, octahedral as well as pentagonal bipyramidal derivatives led to effective inhibitors. However, the five isozymes showed very different affinities for these anion inhibitors. The best hCA I inhibitors were cyanide, dicyanocuprate and dicyanoaurate (K(I)s in the range of 0.5-7.7 microM), whereas the least effective were fluoride and hexafluoroarsenate. The best hCA II inhibitors were cyanide, hexafluoroferrate and tetrachloroplatinate (K(I)s in the range of 0.02-0.51 mM), whereas the most ineffective ones were fluoride, hexafluoroaluminate and chloride. The best hCA IV inhibitors were dicyanocuprate (K(I) of 9.8 microM) and hexacyanoferrate(II) (K(I) of 10.0 microM), whereas the worst ones were tetrafluoroborate and hexafluoroaluminate (K(I)s in the range of 124-126 mM). The most effective hCA V inhibitors were cyanide, heptafluoroniobate and dicyanocuprate (K(I)s in the range of 0.015-0.79 mM), whereas the most ineffective ones were fluoride, chloride and tetrafluoroborate (K(I)s in the range of 143-241 mM). The best hCA IX inhibitors were on the other hand cyanide, heptafluoroniobate and dicyanoargentate (K(I)s in the range of 4 microM-0.33 mM), whereas the worst ones were hexacyanoferrate(III) and hexacyanoferrate(II). PMID- 15780632 TI - Synthesis and in vitro antitumor activity of 4(3H)-quinazolinone derivatives with dithiocarbamate side chains. AB - A series of 4(3H)-quinazolinone derivatives with dithiocarbamate side chains were synthesized and tested for their in vitro antitumor activity against human myelogenous leukemia K562 cells. Among them, (3,4-dihydro-2-methyl-4 oxoquinazolin-6-yl)methyl 4-(4-fluorophenyl)piperazine-1-carbodithioate 8q exhibited significant inhibitory activity against K562 cells with IC(50) value of 0.5 microM. PMID- 15780633 TI - Good oral absorption prediction on non-nucleoside benzothiadiazine dioxide human cytomegalovirus inhibitors using combined chromatographic and neuronal network techniques. AB - The current drugs available against human cytomegalovirus (HCMV) suffer from a number of shortcomings such as toxic side effect, poor bioavailability and/or risk for emergence of drug-resistance virus strains. Due to these limitations, the development of new drugs against HCMV is of great interest. Taking into account the therapeutic potential of benzothiadiazines dioxides (BTD) derivatives, it is most important to know their oral bioavailability because all the current clinical drugs are poorly absorbed. In this work, the utility of CODES neural networks and biopartitioning micellar chromatography (BMC) in predicting pharmacokinetic properties has been used to estimate the oral absorption of BTD derivatives and their efficacy has been verified. The results indicate higher values for BTD derivatives than the currently licensed anti-viral agents. PMID- 15780634 TI - Antitubercular agents. Part 1: synthesis of phthalimido- and naphthalimido-linked phenazines as new prototype antitubercular agents. AB - The preparation and antitubercular properties of a series of phthalimido- and naphthalimido-linked phenazines are described. Some of these new compounds inhibited the growth of Mycobacterium tuberculosis ATCC 27294, Mycobacterium avium ATCC 49601, Mycobacterium intracellulare ATCC 13950 and some clinical isolates. PMID- 15780635 TI - Antitubercular agents. Part 2: new thiolactomycin analogues active against Mycobacterium tuberculosis. AB - Structurally modified analogues of naturally occurring antibiotic thiolactomycin, substituted at 4-position of the thiolactone ring have been prepared and evaluated for their antitubercular activity. Some of the compounds have exhibited potential activity against Mycobacterium tuberculosis. PMID- 15780636 TI - Structure-activity relationships for 2-anilino-6-phenylpyrido[2,3-d]pyrimidin 7(8H)-ones as inhibitors of the cellular checkpoint kinase Wee1. AB - A series of 2-anilino-6-phenylpyrido[2,3-d]pyrimidin-7(8H)-ones were synthesized and evaluated for their inhibitory properties against the non-receptor kinase c Src and the G2/M checkpoint kinase Wee1. Overall, the compounds were 10-100-fold more potent inhibitors of c-Src than Wee1, and variation of substituents on the 6 phenyl ring did not markedly alter this preference. Solubilizing substituents off the 2-anilino ring in many cases increased Wee1 activity, thus lowering this preference to about 10-fold. 5-Alkyl substituted analogs were generally Wee1 selective, but at the expense of absolute potency. PMID- 15780637 TI - Carbonic anhydrase inhibitors: X-ray crystal structure of a benzenesulfonamide strong CA II and CA IX inhibitor bearing a pentafluorophenylaminothioureido tail in complex with isozyme II. AB - N-1-(4-Sulfamoylphenyl)-N-4-pentafluorophenyl-thiosemicarbazide was prepared by the reaction of 4-isothiocyanato-benzenesulfonamide with pentafluorophenyl hydrazine, and proved to be an effective inhibitor of several isozymes of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), such as CA I, II, and IX. Against the physiologically relevant isozymes hCA II and hCA IX, the compound showed inhibition constants in the range of 15-19 nM, whereas it was less effective as a hCA I inhibitor (K(I) of 78 nM). The high-resolution X-ray crystal structure of its adduct with hCA II showed the inhibitor to bind within the hydrophobic half of the enzyme active site, making extensive and strong van der Waals contacts with amino acid residues Gln92, Val121, Phe131, Leu198, Thr200, Pro202, in addition to the coordination of the sulfonamide nitrogen to the Zn(II) ion of the active site, and participation of the SO(2)NH(2) group to a network of hydrogen bonds involving residues Thr199 and Glu106. These results are helpful for the design of better CA II or CA IX inhibitors based on the thioureido benzenesulfonamide motif, with potential applications as anti-glaucoma or anti cancer drugs. PMID- 15780638 TI - Structure-based design of a new class of highly selective aminoimidazo[1,2 a]pyridine-based inhibitors of cyclin dependent kinases. AB - Structure-based design approach was successfully used to guide the evolution of imidazopyridine scaffold yielding new structural class of highly selective inhibitors of cyclin dependent kinases that were able to form a new interaction with an identified residue of the protein, Lys89. Compounds from this series have shown no detectable effect when tested against a representative set of other serine/threonine kinases such as GSK3beta, CAMKII, PKA, PKC alpha,beta,epsilon,gamma. Compound 2i inhibits proliferation in HCT 116 cells in tissue culture. Synthesis, co-crystal structure of CDK2 in complex with compound 2i, and preliminary SAR study are disclosed. PMID- 15780639 TI - Structure-activity relationships of 1'S-1'-acetoxychavicol acetate for inhibitory effect on NO production in lipopolysaccharide-activated mouse peritoneal macrophages. AB - 1'S-1'-Acetoxychavicol acetate from the rhizomes of Alpinia galanga inhibited nitric oxide (NO) production in lipopolysaccharide-activated mouse peritoneal macrophages with an IC(50) value of 2.3 microM. To clarify the structure-activity relationship of 1'S-1'-acetoxychavicol acetate, various natural and synthetic phenylpropanoids and synthetic phenylbutanoids were examined, and the following structural requirements were clarified. (1) The para or ortho substitution of the acetoxyl and 1-acetoxypropenyl groups at the benzene ring was essential. (2) The S configuration of the 1'-acetoxyl group was preferable. (3) The presence of the 3-methoxyl group and disappearance of the 2'-3' double bond by hydrogenation reduced the activity. (4) The substitution of acetyl groups with propionyl or methyl groups reduced the activity. (5) Lengthening of the carbon chain between the 1'- and 2'-positions reduced the activity. PMID- 15780640 TI - Analysis of ADAMTS4 and MT4-MMP indicates that both are involved in aggrecanolysis in interleukin-1-treated bovine cartilage. AB - OBJECTIVE: To investigate the mechanism of aggrecanolysis in interleukin-1 (IL-1) treated cartilage tissue by examining the time course of aggrecan cleavages and the tissue and medium content of membrane type 4-matrix metalloproteinases (MT4 MMP) and a disintegrin and metalloproteinase with thrombospondin type I motifs (ADAMTS)4. METHODS: Articular cartilage explants were harvested from newborn bovine femoropatellar groove. The effects of IL-1 treatment with or without aggrecanase blockade were investigated by Western analysis of aggrecan fragment generation, ADAMTS4 species (p68 and p53), and MT4-MMP, as well as by realtime PCR (polymerase chain reaction) for ADAMTS4 and 5. Aggrecanase was blocked with mannosamine (ManN), an inhibitor of glycosylphosphatidylinositol anchor synthesis, and esculetin (EST), an inhibitor of MMP-1, MMP-3, and MMP-13 gene expression. RESULTS: IL-1 treatment caused a major increase in MT4-MMP abundance in the tissue and medium. ADAMTS4 (p68) was abundant in fresh cartilage and this was retained in the tissue in untreated cartilage. IL-1 treatment for 6 days caused a marked loss of p68 from the cartilage and the appearance of p53 in the medium. Addition of either 1.35 mM ManN or 31-500 microM EST blocked IL-1 mediated aggrecanolysis and this was accompanied by nearly complete inhibition of the MT4-MMP increase, the p68 loss and the formation of p53. IL-1 treatment increased mRNA abundance for ADAMTS4 ( approximately 3-fold) and ADAMTS5 ( approximately 10-fold) but this was not accompanied by a marked change in enzyme protein abundance. CONCLUSION: These studies support a central role for MT4-MMP in IL-1-induced cartilage aggrecanolysis and are consistent with the identification of p68 as the aggrecanase that cleaves within the CS2 domain, and of p53 as the aggrecanase that generates G1-NITEGE. Since the induction by IL-1 was not accompanied by marked changes in total ADAMTS4 protein, but rather in partial conversion of p68 to p53 and release of both from the tissue, we conclude that aggrecanolysis in this model system results from MT4-MMP-mediated processing of a resident pool of ADAMTS4 and release of the p68 and p53 from their normal association with the cell surface. PMID- 15780641 TI - Effects of FGF-2 and IGF-1 on adult canine articular chondrocytes in type II collagen-glycosaminoglycan scaffolds in vitro. AB - OBJECTIVE: Chondrocyte-seeded tissue engineering scaffolds hold the promise of enhancing certain cartilage repair procedures. The objective of this study was to evaluate the effects of selected growth factors [fibroblast growth factor (FGF)-2 and insulin-like growth factor (IGF)-1] individually and in combination on adult canine articular chondrocyte-seeded type II collagen-glycosaminoglycan (GAG) scaffolds grown in serum-free (SF) medium. DESIGN: Approximately 500,000 second passage chondrocytes were seeded into discs of the scaffold, 4mm diameterx2 mm thick. The constructs were grown in the following media: serum-containing medium; a basal SF medium; SF with 5 ng/ml FGF-2; SF with 25 ng/ml FGF-2; SF with 100 ng/ml IGF-1; and SF with 5 ng/ml FGF-2 plus 100 ng/ml IGF-1. The DNA and GAG contents of the scaffolds were determined after 1 day and 2 weeks and the protein and GAG synthesis rates determined at 2 weeks using radiolabels. Histology and type II collagen immunohistochemistry were also performed. RESULTS: FGF-2 at 5 ng/ml was found to substantially increase the biosynthetic activity of the cells and the accumulation of GAG. The histology demonstrated chondrocytes uniformly distributed through a matrix that stained intensely for GAG and type II collagen after only 2 weeks. Of interest were the rapid degradation of the collagen scaffold, despite the fact that the scaffold was carbodiimide cross-linked, and the contraction of the constructs. There were less pronounced effects using the higher dose of FGF-2 and the combination with IGF-1. CONCLUSIONS: Chondrocyte seeded type II collagen scaffolds cultured in SF medium supplemented with 5 ng/ml FGF-2 undergo contraction, demonstrate an increase in construct incorporation of radiolabeled sulfate, and display qualitative signs of chondrogenesis. PMID- 15780642 TI - AG-041R, a novel indoline-2-one derivative, stimulates chondrogenesis in a bipotent chondroprogenitor cell line CL-1. AB - OBJECTIVE: To examine the chondrogenic activity of AG-041R and its mode of action in a bipotent chondroprogenitor cell line CL-1. DESIGN: Chondrogenic activity of AG-041R in CL-1 was examined by histology, alcian blue pH 1.0 intensity and mRNA expression of cartilage matrix proteins (collagen type II, aggrecan). Chondrogenic activities of other CCK2/gastrin receptor antagonists were also examined. Since TGF-beta1 induces dominant chondrogenesis and suppressed adipogenesis in CL-1, induction of TGF-beta by AG-041R was examined by enzyme linked immunosorbent assay. Involvement of MAP kinases in the chondrogenic effect of AG-041R in CL-1 was examined by Western blotting and MAP kinase inhibitors. RESULTS: AG-041R induced dominant chondrogenesis and marked suppression of adipogenesis in CL-1. Neither of the other CCK2/gastrin receptor antagonists tested showed chondrogenic activity in CL-1. AG-041R increased alcian blue pH 1.0 intensity and mRNA expression of collagen type II and aggrecan. TGF-beta1 and beta2 proteins were increased by AG-041R. The chondrogenic activity of AG-041R in CL-1 was blocked by TGF-beta neutralizing antibody or inhibitors for activation of latent TGF-beta. AG-041R activated both Erk (p44/42) and p38 MAP kinases in CL 1. Inhibition of Erk (p44/42) by PD98059 canceled the adipogenesis suppression by AG-041R in CL-1. Inhibition of p38 by SB202190 completely canceled the chondrogenic activity of AG-041R in CL-1. CONCLUSION: AG-041R has chondrogenic activity in CL-1 not related to CCK2/gastrin receptor antagonism. It is suggested that TGF-beta induction and the activation of MAP kinases mediate the chondrogenic activity of AG-041R in CL-1. PMID- 15780643 TI - Repair of osteochondral defects with hyaluronan- and polyester-based scaffolds. AB - OBJECTIVE: The natural repair of osteochondral defects can be enhanced with biocompatible, biodegradable materials that support the repair process. It is our hypothesis that hyaluronan-based scaffolds are superior to synthetic scaffolds because they provide biological cues. We tested this thesis by comparing two hyaluronan-based scaffolds [auto cross-linked polysaccharide polymer (ACP) and HYAFF-11] to polyester-based scaffolds [poly(DL-lactic-co-glycolic acid) (PLGA) and poly(L-lactic acid) (PLLA)] with similar pore size, porosity and degradation times. DESIGN: Fifty-four rabbits received bilateral osteochondral defects. One defect received a hyaluronan-based scaffold and the contralateral defect received the corresponding polyester-based scaffold. Rabbits were euthanized 4, 12 and 20 weeks after surgery and the condyles dissected and processed for histology. RESULTS: Only ACP-treated defects presented bone at the base of the defect at 4 weeks. At 12 weeks, only defects treated with rapidly dissolving implants (ACP and PLGA) presented bone reconstitution consistently, while bone was present in only one third of those treated with slowly dissolving scaffolds (HYAFF-11 and PLLA). After 20 weeks, the articular surface of PLGA-treated defects presented fibrillation more frequently than in ACP-treated defects. The surface of defects treated with slowly dissolving scaffolds presented more cracks and fissures. CONCLUSIONS: The degradation rate of the scaffolds is critical for the repair process. Slowly dissolving scaffolds sustain thicker cartilage at the surface but, it frequently presents cracks and discontinuities. These scaffolds also delay bone formation at the base of the defects. Hyaluronan-based scaffolds appear to allow faster cell infiltration leading to faster tissue formation. The degradation of ACP leads to rapid bone formation while the slow degradation of HYAFF-11 prolongs the presence of cartilage and delays endochondral bone formation. PMID- 15780644 TI - Subchondral and trabecular bone metabolism regulation in canine experimental knee osteoarthritis. AB - OBJECTIVE: To determine trabecular and subchondral bone metabolic changes in experimental canine osteoarthritis (OA). METHODS: OA was induced in 19 dogs by transection of the anterior cruciate ligament (ACL) of the right knee through a stab wound. Dogs were sacrificed at 8 (n=7) and 12 weeks (n=12) after surgery. Non-operated normal dogs (n=6) were used as controls. After sacrifice, samples were obtained from the weight-bearing area of medial tibial plateaus. Explants and cell cultures were prepared from subchondral and trabecular bone. Osteocalcin (Oc), cellular alkaline phosphatase (ALPase), urokinase plasminogen-activator (uPA), prostaglandin E2 (PGE2), metalloproteinase (MMP) and nitric oxide (NO) were measured using standard procedures. RESULTS: ALPase production was significantly increased only at week 12 in subchondral and trabecular bone, while an increase in Oc was noted at week 8. uPA and MMP activity were increased significantly at week 12 in subchondral bone, while PGE2 levels were significantly higher in subchondral and trabecular bone at week 12 compared to normal. A decrease in NO production appeared late at week 12 in trabecular bone, whereas NO levels from subchondral bone were significantly increased compared to normal at week 8. DISCUSSION: Intense bone remodeling takes place in both subchondral and trabecular bone in the knee following ACL transection. This process seems to occur around week 12, although Oc and NO appeared to be involved earlier at 8 weeks. These results suggest that not only subchondral but also trabecular bone metabolism is altered in this OA model. PMID- 15780645 TI - Tissue engineering of cartilage using an injectable and adhesive chitosan-based cell-delivery vehicle. AB - OBJECTIVE: Adult articular cartilage shows a limited intrinsic repair response to traumatic injury. To regenerate damaged cartilage, cell-assisted repair is thus viewed as a promising therapy, despite being limited by the lack of a suitable technique to deliver and retain chondrogenic cells at the defect site. DESIGN: We have developed a cytocompatible chitosan solution that is space-filling, gels within minutes, and adheres to cartilage and bone in situ. This unique combination of properties suggested significant potential for its use as an arthroscopically injectable vehicle for cell-assisted cartilage repair. The primary goal of this study was to assess the ability of this polymer system, when loaded with primary articular chondrocytes, to support cartilage formation in vitro and in vivo. The chitosan gel was cultured in vitro, with and without chondrocytes, as well as injected subcutaneously in nude mice to form subcutaneous dorsal implants. In vitro and in vivo constructs were collectively analyzed histologically, for chondrocyte mRNA and protein expression, for biochemical levels of glycosaminoglycan, collagen, and DNA, and for mechanical properties. RESULTS: Resulting tissue constructs revealed histochemical, biochemical and mechanical properties comparable to those observed in vitro for primary chondrocytes cultured in 2% agarose. Moreover, the gel was retained after injection into a surgically prepared, rabbit full-thickness chondral defect after 1 day in vivo, and in rabbit osteochondral defects, up to 1 week. CONCLUSIONS: The in situ-gelling chitosan solution described here can support in vitro and in vivo accumulation of cartilage matrix by primary chondrocytes, while persisting in osteochondral defects at least 1 week in vivo. PMID- 15780647 TI - MR imaging of articular cartilage at 1.5T and 3.0T: comparison of SPGR and SSFP sequences. AB - OBJECTIVE: To compare articular cartilage signal-to-noise ratio (SNR), contrast to-noise ratio (CNR), and thickness measurements on a 1.5 T and a 3.0 T magnetic resonance (MR) scanner using three-dimensional spoiled gradient recalled echo (3D SPGR) and two 3D steady-state free precession (SSFP) sequences. METHODS: Both knees of five volunteers were scanned at 1.5 T and at 3.0 T using a transmit receive quadrature extremity coil. Each examination consisted of a sagittal 3D SPGR sequence, a sagittal fat suppressed 3D-SSFP (FS-SSFP) sequence, and a sagittal Dixon 3D-SSFP sequence. For quantitative analysis, we compared cartilage SNR and CNR efficiencies, as well as average cartilage thickness measurements. RESULTS: For 3D-SPGR, cartilage SNR efficiencies at 3.0 T increased compared to those at 1.5 T by a factor of 1.83 (range: 1.40-2.09). In comparison to 3D-SPGR, the SNR efficiency of FS-SSFP increased by a factor of 2.13 (range: 1.81-2.39) and for Dixon SSFP by a factor of 2.39 (range: 1.95-2.99). For 3D-SPGR, CNR efficiencies between cartilage and its surrounding tissue increased compared to those at 1.5 T by a factor of 2.12 (range: 1.75-2.47), for FS-SSFP by a factor 2.11 (range: 1.58-2.80) and for Dixon SSFP by a factor 2.39 (range 2.09-2.83). Average cartilage thicknesses of load bearing regions were not different at both field strengths or between sequences (P>0.05). Mean average cartilage thickness measured in all knees was 2.28 mm. CONCLUSION: Articular cartilage imaging of the knee on a 3.0 T MR scanner shows increased SNR and CNR efficiencies compared to a 1.5 T scanner, where SSFP-based techniques show the highest increase in SNR and CNR efficiency. There was no difference between average cartilage thickness measurements performed at the 1.5 T and 3.0 T scanners or between the three different sequences. PMID- 15780646 TI - Human articular chondrocytes exhibit sexual dimorphism in their responses to 17beta-estradiol. AB - OBJECTIVE: The higher incidence of osteoarthritis in females suggests that there may be intrinsic sex-specific differences in human articular chondrocytes. 17beta Estradiol (E2) regulates rat growth plate chondrocytes through traditional nuclear receptor mechanisms, but only female cells exhibit rapid membrane associated effects mediated through protein kinase C (PKC) alpha. Here we demonstrate sexual dimorphism in the physiological response of human articular chondrocytes to E2. METHODS: Articular chondrocytes were obtained at the time of autopsy from three male and three female donors between 16 and 39 years of age. Second passage cultures were treated with E2 for 24 h to assess the effects of the hormone on [3H]-thymidine incorporation, [35S]-sulfate incorporation, and alkaline phosphatase specific activity. In addition, the chondrocytes were treated for 3, 9, 90 or 270 min and PKC specific activity was determined. RESULTS: All chondrocytes were positive for aggrecan and estrogen receptor alpha mRNAs but were negative for type II collagen mRNA. Only cells from female donors responded to E2. DNA synthesis, sulfate incorporation and alkaline phosphatase activity were increased. E2 caused a rapid increase in PKC activity in the female cells within 9 min that was maximal at 90 min. Treatment with the PKC inhibitor chelerythrine blocked these effects. CONCLUSIONS: These results provide the first definitive evidence that normal human cells exhibit an intrinsic sex-specific response to E2 and suggest that sexual dimorphism may be an important variable in assessing the pathways that modulate cell behavior. PMID- 15780648 TI - Nonlinear optical microscopy of articular cartilage. AB - OBJECTIVE: To assess the ability of nonlinear optical microscopy (NLOM) to image ex vivo healthy and degenerative bovine articular cartilage. METHOD: Fresh bovine femoral-tibial joints were obtained from an abattoir. Articular cartilage specimens were harvested from the tibial plateau. Normal and degenerative specimens were imaged by NLOM and subsequently fixed and processed for histological examination. RESULTS: NLOM provided high resolution images of articular cartilage at varying depths with high sensitivity to tissue morphology and high specificity to tissue components without fixing, sectioning or staining. Spectroscopic segmentation of nonlinear optical signals isolated the collagen matrix from the chondron (chondrocyte and non-collagen pericellular matrix). Images from the superficial zone were consistent with the presence of a matrix composed of both elastin-like and collagen fibers distributed in a depth dependent morphological arrangement, whereas only collagen was demonstrated in the middle and deep zones. Alterations of collagen matrix associated with advanced degenerative joint disease (fibrocartilage) were observed with NLOM. Individual chondrocytes were imaged and demonstrated intracellular fluorescence consistent with the presence of products of intracellular biochemical processes. CONCLUSION: Thin images of living articular cartilage using NLOM may be obtained with (sub-)cellular resolution at varying depths without fixing, sectioning or staining. Extracellular matrical collagen and chondron may be imaged separately in native tissue using spectrally distinct, endogenous, nonlinear optical signals. NLOM was sensitive to macromolecular composition and pathologic changes in articular cartilage matrix. Advances in instrumentation may lead to the application of NLOM to study articular cartilage in vivo. PMID- 15780649 TI - An optimal duration of daily wear for an insole with subtalar strapping in patients with varus deformity osteoarthritis of the knee. AB - OBJECTIVE: To assess the optimal duration of daily wear for a laterally wedged insole with subtalar strapping in subjects with medial compartment osteoarthritis of the knee (knee OA). DESIGN: The setting was an outpatient clinic. Eighty-one patients with knee OA were prospectively randomized according to birth date and to either 2 weeks of treatment with a lateral wedge with subtalar strapping for less than 5 h (the short group), 5-10 h (the medium group) or greater than 10 h (the long group) each day, or to treatment with a subtalar strapping band without lateral wedge (the placebo group). Standing radiographs were used to analyze the femorotibial angle for each subject, both with and without their respective orthotic device. The remission scores of Lequesne index were compared among the four groups at the conclusion. RESULTS: The short (n=21), medium (n=20) and long (n=18) groups demonstrated a significant greater valgus correction of the femorotibial angle than the placebo group (n=22) (P<0.0001). The remission score was significantly improved in the medium group compared to the placebo (P=0.001) and long (P=0.001) groups. CONCLUSIONS: An optimal duration of insole with subtalar strapping wear for patients with varus deformity knee OA may be between 5 and 10 h each day. PMID- 15780651 TI - Acid trehalase in yeasts and filamentous fungi: localization, regulation and physiological function. AB - Yeasts and filamentous fungi are endowed with two different trehalose-hydrolysing activities, termed acid and neutral trehalases according to their optimal pH for enzymatic activity. A wealth of information already exists on fungal neutral trehalases, while data on localization, regulation and function of fungal acid trehalases have remained elusive. The gene encoding the latter enzyme has now been isolated from two yeast species and two filamentous fungi, and sequences encoding putative acid trehalase can be retrieved from available public sequences. Despite weak similarities between amino acids sequences, this type of trehalase potentially harbours either a transmembrane segment or a signal peptide at the N-terminal sequence, as deduced from domain prediction algorithms. This feature, together with the demonstration that acid trehalase from yeasts and filamentous fungi is localized at the cell surface, is consistent with its main role in the utilisation of exogenous trehalose as a carbon source. The growth on this disaccharide is in fact pretty effective in most fungi except in Saccharomyces cerevisiae. This yeast species actually exhibits a "Kluyver effect" on trehalose. Moreover, an oscillatory behaviour reminiscent of what is observed in aerobic glucose-limited continuous cultures at low dilution rate is also observed in batch growth on trehalose. Finally, the S. cerevisiae acid trehalase may also participate in the catabolism of endogenous trehalose by a mechanism that likely requires the export of the disaccharide, its extracellular hydrolysis, and the subsequent uptake of the glucose released. Based on these recent findings, we suggest to rename "acid" and "neutral" trehalases as "extracellular" and "cytosolic" trehalases, which is more adequate to describe their localization and function in the fungal cell. PMID- 15780652 TI - Strain engineering for stereoselective bioreduction of dicarbonyl compounds by yeast reductases. AB - Pure chiral molecules are needed in the pharmaceutical and chemical industry as intermediates for the production of drugs or fine chemicals. Microorganisms represent an attractive alternative to chemical synthesis since they have the potential to generate single stereoisomers in high enantiomeric excess (ee). The baker's yeast Saccharomyces cerevisiae can notably reduce dicarbonyl compounds (in particular alpha- and beta-diketones and keto esters) to chiral alcohols with high ee. However, products are formed at a low rate. Moreover, large amounts of co-substrate are required for the regeneration of NADPH that is the preferred co factor in almost all the known dicarbonyl reductions. Traditionally, better ee, reduction rate and product titre have been achieved via process engineering. The advent of recombinant DNA technology provides an alternative strategy to improve productivity and yield by strain engineering. This review discusses two aspects of strain engineering: (i) the generation of strains with higher reductase activity towards dicarbonyl compounds and (ii) the optimisation of co-substrate utilisation for NADPH cofactor regeneration. PMID- 15780653 TI - Hydrophobic substrate utilisation by the yeast Yarrowia lipolytica, and its potential applications. AB - The alkane-assimilating yeast Yarrowia lipolytica degrades very efficiently hydrophobic substrates such as n-alkanes, fatty acids, fats and oils for which it has specific metabolic pathways. An overview of the oxidative degradation pathways for alkanes and triglycerides in Y. lipolytica is given, with new insights arising from the recent genome sequencing of this yeast. This includes the interaction of hydrophobic substrates with yeast cells, their uptake and transport, the primary alkane oxidation to the corresponding fatty alcohols and then by different enzymes to fatty acids, and the subsequent degradation in peroxisomal beta-oxidation or storage into lipid bodies. Several enzymes involved in hydrophobic substrate utilisation belong to multigene families, such as lipases/esterases (LIP genes), cytochromes P450 (ALK genes) and peroxisomal acyl CoA oxidases (POX genes). Examples are presented demonstrating that wild-type and genetically engineered strains of Y. lipolytica can be used for alkane and fatty acid bioconversion, such as aroma production, for production of SCP and SCO, for citric acid production, in bioremediation, in fine chemistry, for steroid biotransformation, and in food industry. These examples demonstrate distinct advantages of Y. lipolytica for their use in bioconversion reactions of biotechnologically interesting hydrophobic substrates. PMID- 15780654 TI - Metabolic-flux and network analysis in fourteen hemiascomycetous yeasts. AB - In a quantitative comparative study, we elucidated the glucose metabolism in fourteen hemiascomycetous yeasts from the Genolevures project. The metabolic networks of these different species were first established by (13)C-labeling data and the inventory of the genomes. This information was subsequently used for metabolic-flux ratio analysis to quantify the intracellular carbon flux distributions in these yeast species. Firstly, we found that compartmentation of amino acid biosynthesis in most species was identical to that in Saccharomyces cerevisiae. Exceptions were the mitochondrial origin of aspartate biosynthesis in Yarrowia lipolytica and the cytosolic origin of alanine biosynthesis in S. kluyveri. Secondly, the control of flux through the TCA cycle was inversely correlated with the ethanol production rate, with S. cerevisiae being the yeast with the highest ethanol production capacity. The classification between respiratory and respiro-fermentative metabolism, however, was not qualitatively exclusive but quantitatively gradual. Thirdly, the flux through the pentose phosphate (PP) pathway was correlated to the yield of biomass, suggesting a balanced production and consumption of NADPH. Generally, this implies the lack of active transhydrogenase-like activities in hemiascomycetous yeasts under the tested growth condition, with Pichia angusta as the sole exception. In the latter case, about 40% of the NADPH was produced in the PP pathway in excess of the requirements for biomass production, which strongly suggests the operation of a yet unidentified mechanism for NADPH reoxidation in this species. In most yeasts, the PP pathway activity appears to be driven exclusively by the demand for NADPH. PMID- 15780655 TI - Metabolic-flux analysis of Saccharomyces cerevisiae CEN.PK113-7D based on mass isotopomer measurements of (13)C-labeled primary metabolites. AB - Metabolic-flux analyses in microorganisms are increasingly based on (13)C labeling data. In this paper a new approach for the measurement of (13)C-label distributions is presented: rapid sampling and quenching of microorganisms from a cultivation, followed by extraction and detection by liquid chromatography-mass spectrometry of free intracellular metabolites. This approach allows the direct assessment of mass isotopomer distributions of primary metabolites. The method is applied to the glycolytic and pentose phosphate pathways of Saccharomyces cerevisiae strain CEN.PK113-7D grown in an aerobic, glucose-limited chemostat culture. Detailed investigations of the measured mass isotopomer distributions demonstrate the accuracy and information-richness of the obtained data. The mass fractions are fitted with a cumomer model to yield the metabolic fluxes. It is estimated that 24% of the consumed glucose is catabolized via the pentose phosphate pathway. Furthermore, it is found that turnover of storage carbohydrates occurs. Inclusion of this turnover in the model leads to a large confidence interval of the estimated split ratio. PMID- 15780656 TI - Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae. AB - Various selection procedures in chemostats and batch cultures were systematically tested for their efficiency to select for a multiple-stress resistance phenotype in Saccharomyces cerevisiae. To determine the relative stress resistance phenotypes, mutant populations harvested at different time points and randomly chosen clones from selected populations were grown in batch cultures and exposed to oxidative, freezing-thawing, high-temperature and ethanol stress. For this purpose, we developed a high-throughput procedure in 96-well plates combined with a most-probable-number assay. Among all chemostat and batch selection strategies tested, the best selection strategy to obtain highly improved multiple-stress resistant yeast was found to be batch selection for freezing-thawing stress. The final mutant populations selected for this particular stress were not only significantly improved in freezing-thawing stress resistance, but also in other stress resistances. The best isolated clone from these populations exhibited 102 , 89-, 62-, and 1429-fold increased resistance to freezing-thawing, temperature, ethanol, and oxidative stress, respectively. General selection guidelines for improving multiple-stress resistance in S. cerevisiae are presented and discussed. PMID- 15780657 TI - Physiological and genome-wide transcriptional responses of Saccharomyces cerevisiae to high carbon dioxide concentrations. AB - Physiological effects of carbon dioxide and impact on genome-wide transcript profiles were analysed in chemostat cultures of Saccharomyces cerevisiae. In anaerobic, glucose-limited chemostat cultures grown at atmospheric pressure, cultivation under CO(2)-saturated conditions had only a marginal (<10%) impact on the biomass yield. Conversely, a 25% decrease of the biomass yield was found in aerobic, glucose-limited chemostat cultures aerated with a mixture of 79% CO(2) and 21% O(2). This observation indicated that respiratory metabolism is more sensitive to CO(2) than fermentative metabolism. Consistent with the more pronounced physiological effects of CO(2) in respiratory cultures, the number of CO(2)-responsive transcripts was higher in aerobic cultures than in anaerobic cultures. Many genes involved in mitochondrial functions showed a transcriptional response to elevated CO(2) concentrations. This is consistent with an uncoupling effect of CO(2) and/or intracellular bicarbonate on the mitochondrial inner membrane. Other transcripts that showed a significant transcriptional response to elevated CO(2) included NCE103 (probably encoding carbonic anhydrase), PCK1 (encoding PEP carboxykinase) and members of the IMD gene family (encoding isozymes of inosine monophosphate dehydrogenase). PMID- 15780658 TI - Intraspecific gene expression variability in the yeast Kluyveromyces lactis revealed by micro-array analysis. AB - Using the Genolevures sequencing data, we developed an expression micro-array for the yeast Kluyveromyces lactis consisting of 482 genes, mainly involved in central metabolism, compound transport facilitators and stress response. The array was validated using the LAC/GAL system. By comparing gene expression in the laboratory reference strain CBS2359 and in an industrial strain B1, we demonstrated the influence of two carbon sources, glucose and lactose, on the expression of genes involved in the respiratory and in the fermentative metabolic pathways. We also showed that the two strains, although both originating from dairies, display unexpected differences in gene expression on each type of carbon source. PMID- 15780659 TI - Gln3p and Nil1p regulation of invertase activity and SUC2 expression in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, sensing and signalling pathways regulate gene expression in response to quality of carbon and nitrogen sources. One such system, the target of rapamycin (Tor) proteins, senses nutrients and uses the GATA activators Gln3p and Nil1p to regulate translation in response to low quality carbon and nitrogen. The signal transduction, triggered in response to nitrogen nutrition that is sensed by the Tor proteins, operates via a regulatory pathway involving the cytoplasmic factor Ure2p. When carbon and nitrogen are abundant, the phosphorylated Ure2p anchors the also phosphorylated Gln3p and Nil1p in the cytoplasm. Upon a shift from high- to low-quality nitrogen or treatment with rapamycin all three proteins are dephosphorylated, causing Gln3p and Nil1p to enter the nucleus and promote transcription. The genes that code for yeast periplasmic enzymes with nutritional roles would be obvious targets for regulation by the sensing and signalling pathways that respond to quality of carbon and nitrogen sources. Indeed, previous results from our laboratory had shown that the GATA factors Gln3p, Nil1p, Dal80p, Nil2p and also the protein Ure2 regulate the expression of asparaginase II, coded by ASP3. We also had observed that the activity levels of the also periplasmic invertase, coded by SUC2, were 6 fold lower in ure2 mutant cells in comparison to wild-type cells collected at stationary phase. These results suggested similarities between the signalling pathways regulating the expression of ASP3 and SUC2. In the present work we showed that invertase levels displayed by the single nil1 and gln3 and by the double gln3nil1 mutant cells, cultivated in a sucrose-ammonium medium and collected at the exponential phase, were 6-, 10- and 60-fold higher, respectively, in comparison to their wild-type counterparts. RT-PCR data of SUC2 expression in the double-mutant cells indicated a 10-fold increase in the mRNA(SUC2) levels. PMID- 15780660 TI - Hierarchical and metabolic regulation of glucose influx in starved Saccharomyces cerevisiae. AB - A novel method dissecting the regulation of a cellular function into direct metabolic regulation and hierarchical (e.g., gene-expression) regulation is applied to yeast starved for nitrogen or carbon. Upon nitrogen starvation glucose influx is down-regulated hierarchically. Upon carbon starvation it is down regulated both metabolically and hierarchically. The method is expounded in terms of its implications for diverse types of regulation. It is also fine-tuned for cases where isoenzymes catalyze the flux through a single metabolic step. PMID- 15780661 TI - Efficient conversion of 11-deoxycortisol to cortisol (hydrocortisone) by recombinant fission yeast Schizosaccharomyces pombe. AB - Genetically engineered microorganisms are being increasingly used for the industrial production of complicated chemical compounds such as steroids; however, there have been few reports on the use of the fission yeast Schizosaccharomyces pombe for this purpose. We previously have demonstrated that this yeast is a unique host for recombinant expression of human CYP11B2 (aldosterone synthase), and here we report the functional production of human CYP11B1 (steroid 11beta-hydroxylase) in S. pombe using our new integration vector pCAD1. In the human adrenal, the mitochondrial cytochrome P450 enzyme CYP11B1 catalyses the conversion of 11-deoxycortisol to cortisol, a key reaction in cortisol biosynthesis that in addition is of fundamental interest for the technical synthesis of glucocorticoids. We observed that the endogenous mitochondrial electron transport system detected previously by us is capable of supplying this enzyme with the reducing equivalents necessary for steroid hydroxylation activity. Under optimised cultivation conditions the transformed yeasts show in vivo the inducible ability to efficiently and reliably convert deoxycortisol to cortisol at an average rate of 201 microM d(-1) over a period of 72h, the highest value published to date for this biotransformation. PMID- 15780662 TI - The human mineralocorticoid receptor only partially differentiates between different ligands after expression in fission yeast. AB - Cardiac failure is a major health problem with increasing incidence due to aging of the population. Studies in both experimental animals and humans have suggested that aldosterone excess may have deleterious effects on cardiac function. In order to generate a novel screening system for the identification of aldosterone antagonists, we expressed the human mineralocorticoid receptor (MR) and the human glucocorticoid receptor (GR), respectively, in the fission yeast Schizosaccharomyces pombe. Reporter plasmids containing two hormone-responsive elements upstream of a fission yeast minimal promotor and either a lacZ gene (for quantification) or a neomycin gene (for survival screening) were constructed and cotransformed into fission yeast strains with expression plasmids for MR or GR. The functionality of the reporter systems was then tested using physiological ligands of both receptors as well as known inhibitors. Transactivating activity of MR could be stimulated by aldosterone, 11-deoxycorticosterone, 11 deoxycortisol, cortisol, cortisone, and spironolactone, but not by progesterone, while GR activity was stimulated by cortisol and cortisone, but also not by progesterone. Taken together, we have succeeded in establishing fission yeast based screening systems that allow the identification of MR- or GR-interacting compounds. Moreover, our data show that after expression in fission yeast both receptors did not differentiate between steroids with different configurations at positions 11beta, 17 and 18. This finding suggests that only recognition of C-21 substituents may be accomplished by the receptor proteins alone, while the physiologically important selectivity towards other positions of the steroid ligand depends on other factors which are not conserved from fission yeast to man. PMID- 15780663 TI - Heterologous production of a laccase from the basidiomycete Pycnoporus cinnabarinus in the dimorphic yeast Yarrowia lipolytica. AB - Pycnoporus cinnabarinus lac1 gene was expressed in Yarrowia lipolytica. Different secretion signals and culture media were tested. Production was correlated to both culture growth rate and cell morphology (highest at low growth rate, without mycelium). Recombinant laccase was characterized (immunodetection, N-terminal sequencing) and purified. Production was estimated to 20 mgl(-1) in a bioreactor. Thus, complex metalloenzymes can be produced in Yarrowia, assuming some control of host physiology. Lac1p production was compared in Yarrowia, Pichia and Aspergillus: recombinant proteins were active, but host systems differed in transformation efficiency, production, and glycosylation. If not the best producer, Yarrowia offers very high transformation efficiencies, allowing the genetic engineering of laccases for industrial applications. PMID- 15780664 TI - Heterologous protein production in Zygosaccharomyces bailii: physiological effects and fermentative strategies. AB - The optimisation and scale-up of a specific protein production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this "non-conventional" yeast suitable for the development of biotechnological processes like heterologous protein production. This work addresses the production of human interleukin-1beta by a recombinant Z. bailii strain. We found that the heterologous protein production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1beta production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of production and productivity was the fed-batch mode. PMID- 15780665 TI - Development of a highly efficient gene targeting system for Fusarium graminearum using the disruption of a polyketide synthase gene as a visible marker. AB - We cloned a polyketide synthase gene (pks12) from Fusarium graminearum, a devastating fungal pathogen of cereals. Transformation-mediated gene disruption led to an easily detectable albino phenotype of the disruptants. We used the disruption of the pks12 gene as a visible marker for transformation-mediated homologous recombination and optimized the transformation procedure to achieve a high rate of homologous recombination. In combination with the published genomic sequence data and the generation of expressed sequence tags (ESTs) for F. graminearum, this is a useful tool to investigate this important plant pathogen on a molecular level. Optimized transformation of F. graminearum resulted in at least 93% homologous recombination events when the homologous genomic DNA fragment in the vector had a size of approximately 800bp and was linearized in the middle. Using a genomic sequence of approximately 500bp in the transformation vector, 70% of the transformants still exhibited homologous recombination. On the contrary, no more than 10% homologous recombination events were observed when less than 400bp DNA fragments were used. We co-transformed F. graminearum with two different vectors. One vector harboured a DNA insert homologous to the pks12 gene, while the other vector consisted of the same vector backbone carrying the selection marker specific for F. graminearum. About 70% of the transformants had a disrupted pks12 gene, and all of these showed an integration of the second vector into the pks disruption vector. Therefore, the time-consuming construction of a single transformation vector can be avoided; furthermore, it is now easily feasible to express a gene construct at a defined and mutated genomic site. PMID- 15780666 TI - Endo-polygalacturonase in Saccharomyces wine yeasts: effect of carbon source on enzyme production. AB - Eight wine yeast strains of Saccharomyces sp. were tested for polygalacturonase (PGase) activity, after cultivation on various carbon sources. No strain showed any activity when grown on glucose, while five strains produced PGase in the presence of galactose and polygalacturonate. These data suggest that the PGase of wine strains is repressed by glucose and induced by galactose and polygalacturonate. The existence of the PGase gene in the wine strains and its similarity with that of the laboratory strains was proved by Southern hybridization and PCR amplification. The promoter region of the PGase gene in the wine strains was slightly different from that of the laboratory strains. This possibly explains the different pattern of gene expression in wine and laboratory strains. The PGase of wine strains produced di- or tri-galacturonic acid from polygalacturonic acid, different from the fungal PGase. PMID- 15780668 TI - Amino acid supplementation, controlled oxygen limitation and sequential double induction improves heterologous xylanase production by Pichia stipitis. AB - Heterologous endo-beta-1,4-xylanase was produced by Pichia stipitis under control of the hypoxia-inducible PsADH2-promoter in a high-cell-density culture. After promoter induction by a shift to oxygen limitation, different aeration rates (oxygen transfer rates) were applied while maintaining oxygen-limitation. Initially, enzyme production was higher in oxygen-limited cultures with high rates of oxygen transfer, although the maximum xylanase activity was not significantly influenced. Amino acid supplementation increased the production of the heterologous endo-beta-1,4-xylanase significantly in highly aerated oxygen limited cultures, until glucose was depleted. A slight second induction of the promoter was observed in all cultures after the glucose had been consumed. The second induction was most obvious in amino acid-supplemented cultures with higher oxygen transfer rates during oxygen limitation. When such oxygen-limited cultures were shifted back to fully aerobic conditions, a significant re-induction of heterologous endo-beta-1,4-xylanase production was observed. Re-induction was accompanied by ethanol consumption. A similar protein production pattern was observed when cultures were first grown on ethanol as sole carbon source and subsequently glucose and oxygen limitation were applied. Thus, we present the first expression system in yeast with a sequential double-inducible promoter. PMID- 15780667 TI - Ethanol production from hexoses, pentoses, and dilute-acid hydrolyzate by Mucor indicus. AB - Consumption of hexoses and pentoses and production of ethanol by Mucor indicus were investigated in both synthetic media and dilute-acid hydrolyzates. The fungus was able to grow in a poor medium containing only carbon, nitrogen, phosphate, potassium, and magnesium sources. However, the cultivation took more than a week and the ethanol yield was only 0.2 gg(-1). Enrichment of the medium by addition of trace metals, particularly zinc and yeast extract, improved the growth rate and yield, such that the cultivation was completed in less than 24 h and the ethanol and biomass yields were increased to 0.40 and 0.20 gg(-1), respectively. The fungus was able to assimilate glucose, galactose, mannose, and xylose, and produced ethanol with yields of 0.40, 0.34, 0.39, and 0.18 gg(-1), respectively. However, arabinose was poorly consumed and no formation of ethanol was detected. Glycerol was the major by-product in the cultivation on the hexoses, while formation of glycerol and xylitol were detected in the cultivation of the fungus on xylose. The fungus was able to take up the sugars present in dilute-acid hydrolyzate as well as the inhibitors, acetic acid, furfural, and hydroxymethyl furfural. M. indicus was able to grow under anaerobic conditions when glucose was the sole carbon source, but not on xylose or the hydrolyzate. The yield of ethanol in anaerobic cultivation on glucose was 0.46 g g(-1). PMID- 15780669 TI - Oxoester oxidoreductase activities in new isolates of Pichia anomala from apple, grape and cane juices. AB - Thirty-nine yeasts isolated from apple, grape and cane juices were screened for their oxidoreductase activity. The two strains of Pichia, one isolated from apple and one from cane juices, appear to be promising strains for oxidoreductase activity on alpha-oxoesters. They showed similar high yields in converting ethyl pyruvate to ethyl lactate as Saccharomyces spp. (86.6% and 85.3% versus 86.6%), and higher yields in the reduction of alpha-oxocarboxylic esters (ketopantolactone to pantolactone: 74% and 73.3%, respectively) compared to Saccharomyces spp. (yield 60%). PMID- 15780670 TI - TEMPS: Temperament Evaluation of Memphis, Pisa, Paris and San Diego. PMID- 15780671 TI - TEMPS-A: progress towards validation of a self-rated clinical version of the Temperament Evaluation of the Memphis, Pisa, Paris, and San Diego Autoquestionnaire. AB - BACKGROUND: Our aim was to validate the Temperament Evaluation of the Memphis, Pisa, Paris, and San Diego Autoquestionnaire (TEMPS-A) in a clinical population. METHODS: The study was conducted in two Memphis mood clinics involving 398 affectively ill patients with young to middle index age (42 years+/-13 S.D.), who were 95% white, 62% female, and 51% bipolar spectrum. A subset of 157 of the entire sample were retested in 6-12 months, and the entire sample was then subjected to factor analysis (PCA extraction method with varimax rotation). RESULTS: We obtained high test-retest reliability ranging from 0.58 for the irritable, to 0.68, 0.69 and 0.70, respectively, for the cyclothymic, dysthymic and hyperthymic. The hypothesized four-factor structure of the TEMPS-A was upheld, with the cyclothymic explaining 14% of the variance, followed by the irritable, hyperthymic, and dysthymic together accounting for another 14%. Internal consistency was excellent, with Chronbach alphas ranging from 0.76 for the dysthymic to 0.88 for the cyclothymic. Exploratory factor analysis revealed 2 super factors, Factor I loading on cyclothymic, irritable, and dysthymic temperaments, and Factor II loading heavily on the hyperthymic. The 50-item TEMPS A-Clinical Version was constructed by using a cutoff of alpha > or =0.4 for traits loading exclusively on their original temperaments. We also proposed a longer 69-item version for future study, in which we permitted a greater number of traits based on clinical considerations (alpha cutoff 0.30). LIMITATION: The sample was preponderantly white, and may not generalize to other U.S. ethnic groups. This earlier version of TEMPS-A did not include the anxious temperament. CONCLUSIONS: We psychometrically validated the TEMPS-A in affectively ill outpatients, leading to an instrument suitable for use in psychiatric, especially affectively ill, populations. It is noteworthy that in this clinically ill population we succeeded in measuring traits which could make subjects vulnerable to affective episodes, as well as those of adaptive nature. For instance, the dysthymic emerged as bound to routine, self-blaming, shy-nonassertive, sensitive to criticism, yet self-denying, dependable, and preferring to work for someone else rather than be the boss. The hyperthymic had the highest number of "positive" traits: upbeat, fun-loving, outgoing, jocular, optimistic, confident, full of ideas, eloquent, on the go, short-sleeper, tireless, who likes to be the boss, but single-minded, risk-taker, and unlikely to admit to his/her meddlesome nature. The cyclothymic emerged as labile with rapid shifts in mood; unstable in energy, self-esteem and socialization; unevenly gifted and dilettante; yet keen in perception, intense in emotions, and romantic. The irritable emerged as skeptical and critical (which might be considered intellectual virtues), but otherwise having the "darkest" nature of all temperaments: grouchy, complaining, dissatisfied; anger- and violence-prone, and sexually jealous. The foregoing temperament attributes, observed in a moderately severe group of patients with affective disorders, nonetheless testify to the evolutionary context of these disorders-"submissive" behavior, territoriality, romantic charm, and last, but not least, sexually jealous with its associated specter of violence. We hypothesize that the putative social and limbic mechanisms underlying mood disorders appear to have archaic origins on an evolutionary scale. We finally submit that the traits underlying affective disorders are very much part of human nature. PMID- 15780672 TI - The relationship of Kraepelian affective temperaments (as measured by TEMPS-I) to the tridimensional personality questionnaire (TPQ). AB - BACKGROUND: There is considerable uncertainty in the current literature about the relationship between personality dimensions and affective temperaments. METHOD: We compared-in a non-ill 14-26-year-old Italian student population of 1010-the affective temperaments of classic psychiatry conceived as subaffective traits [and measured through the Temperament Assessment of Memphis, Pisa, Paris and San Diego-Interview Version (TEMPS-I) in the Akiskal and Mallya Operationalization] with Cloninger's revised Tridimensional Personality Questionnaire (TPQ) deriving from the experimental psychology tradition. RESULTS: The Depressive Temperament (DT) and Harm Avoidance (HA), loaded positively on the same canonical variate, whereas the hyperthymic (HT) strongly, and Novelty Seeking (NS) moderately, loaded negatively. In contrast, the Cyclothymic Temperament (CT) loaded highly positively on a second variate, on which both Novelty Seeking strongly and Harm Avoidance moderately loaded positively. Reward Dependence (RD), Persistence (P), and Irritable Temperament (IT) did not significantly relate to any temperamental and personality constructs. At a subdimensional level of TPQ 'shyness with strangers', 'stoic rigidity', 'detachment', 'fear of uncertainty', 'reflection', and 'anticipatory worry' correlated best with the DT. 'Gregariousness', 'exploratory excitability', 'uninhibited optimism', 'attachment', 'confidence', 'extravagance', 'independence', 'vigor', and 'impulsiveness' correlated best with HT. Lastly, 'anticipatory worry', 'disorderliness', 'sentimentality', and 'fatigability' correlated best with CT. CONCLUSIONS: The data provide concurrent validity to TEMPS-I and, as earlier suggested by Cloninger, indicate that (as expected) high HA and DT are related. High NS is both related to the HT and CT, and (somewhat unexpectedly), the CT is related to high HA. In a more theoretical vein, hyperthymic-novelty seeker can be predicted to be overrepresented among those with high achievement; on the other hand, a moody, restless disposition (a cyclothymic-harm avoidant type) may engage in outrageous behavior and be liable to negative affective arousal. We submit that these considerations could shed some light on the origin of socially adaptive behavior ('sunny' or sanguine types) on the one hand, and borderline conditions, anxious-hostile bipolarity ('dark' types) on the other. PMID- 15780673 TI - Validating affective temperaments in their subaffective and socially positive attributes: psychometric, clinical and familial data from a French national study. AB - BACKGROUND: One of the major objectives of the French National EPIDEP Study was to show the feasibility of systematic assessment of bipolar II (BP-II) disorder and beyond. In this report we focus on the utility of the affective temperament scales (ATS) in delineating this spectrum in its clinical as well as socially desirable expressions. METHODS: Forty-two psychiatrists working in 15 sites in four regions of France made semi-structured diagnoses based on DSM IV criteria in a sample of 452 consecutive major depressive episode (MDE) patients (from which bipolar I had been removed). At least 1 month after entry into the study (when the acute depressive phase had abated), they assessed affective temperaments by using a French version of the precursor of the Temperament Evaluation of Memphis, Pisa, Paris and San Diego (TEMPS). Principal component analyses (PCA) were conducted on hyperthymic (HYP-T), depressive (DEP-T) and cyclothymic (CYC-T) temperament subscales as assessed by clinicians, and on a self-rated cyclothymic temperament (CYC-TSR). Scores on each of the temperament subscales were compared in unipolar (UP) major depressive disorder versus BP-II patients, and in the entire sample subdivided on the basis of family history of bipolarity. RESULTS: PCAs showed the presence of a global major factor for each clinician-rated subscale with respective eigenvalues of the correlation matrices as follows: 7.1 for HYP-T, 6.0 for DEP-T, and 4.7 for CYC-T. Likewise, on the self-rated CYC-TSR, the PCA revealed one global factor (with an eigenvalue of 6.6). Each of these factors represented a melange of both affect-laden and adaptive traits. The scores obtained on clinician and self-ratings of CYC-T were highly correlated (r=0.71). The scores of HYP-T and CYC-T were significantly higher in the BP-II group, and DEP-T in the UP group (P<0.001). Finally, CYC-T scores were significantly higher in patients with a family history of bipolarity. CONCLUSION: These data uphold the validity of the affective temperaments under investigation in terms of face, construct, clinical and family history validity. Despite uniformity of depressive severity at entry into the EPIDEP study, significant differences on ATS assessment were observed between UP and BP-II patients in this large national cohort. Self-rating of cyclothymia proved reliable. Adding the affective temperaments-in particular, the cyclothymic-to conventional assessment methods of depression, a more enriched portrait of mood disorders emerges. More provocatively, our data reveal socially positive traits in clinically recovering patients with mood disorders. PMID- 15780674 TI - Toward a validation of a tripartite concept of a putative anxious temperament: psychometric data from a French national general medical practice study. AB - BACKGROUND: Although generalized anxiety disorder (GAD) is currently described as a time-limited state mental disorder, emerging evidence suggests that it is best considered as an exaggeration of a putative "anxious temperament" (AT). It is presently unknown whether it is a distinct or unitary construct of a melange of anxious traits related to Cluster-C personality disorders. METHODS: As part of a Franco-American collaborative study, we developed the 15-item Operational Criteria for Anxious Personality (OCAP), expanding criteria sets developed earlier by one of us (H.S.A.). The study, which was conducted in the French primary care medical sector, included 1112 young adults (18-40 years), seeking help for isolated anxious complaints, never treated before-and without any diagnosable disorder on the axis I of DSM-IV. As previous papers have reported the preliminary validity of OCAP, especially concurrent validity with the State Trait Anxiety Inventory (STAI) (Speilberger), in this report, we focus on its full psychometric properties. RESULTS: The present data indicate a normal distribution of AT items, a satisfactory Chronbach's coefficient (0.64), and the presence at intake of three different subtypes of AT: "anxious-avoidant," "anxious-phobic," and "anxious-sensitive." After a prospective 6-month follow-up, the major criteria of AT were stable in 80% of cases, and for specific AT items, the stability rate varied between 65% and 80%; much of the unstable items were accounted by improvement during naturalistic treatment. The latter could explain the different factor structure obtained at follow-up, which tended to be less heterogeneous, and represented by one global factor. LIMITATION: We used a categorical (yes/no) rather than a Likert-type gradation of frequency and intensity of anxiousness items and relatively low number of items, especially for those involving worrying about one's own health or that of one's loved ones. CONCLUSIONS: Anxiousness as a temperamental dimension appears to involve putative subtypes along "worrying," "phobic," "sensitive" (and "avoidant") dimensions. PMID- 15780675 TI - TEMPS-A: validation of a short version of a self-rated instrument designed to measure variations in temperament. AB - OBJECTIVE: To validate a short English-language version of the Temperament Evaluation of Memphis, Pisa, Paris and San Diego-autoquestionnaire version (TEMPS A), a self-report questionnaire designed to measure temperamental variations in psychiatric patients and healthy volunteers. Its constituent subscales and items were formulated on the basis of the diagnostic criteria for affective temperaments (cyclothymic, dysthymic, irritable, hyperthymic, and anxious), originally developed by the first author and his former collaborators. Further item wording and selection were achieved at a later stage through an iterative process that incorporated feedback from clinicians, researchers, and research volunteers. METHOD: A total of 510 volunteers (284 patients with mood disorders, 131 relatives of bipolar probands, and 95 normal controls) were recruited by advertisement in the newspapers, announcements on radio and television, flyers and newsletters, and word of mouth. All participants were interviewed using the Structured Clinical Interview for DSM-III-R, and completed the 110-item TEMPS-A and the Temperament and Character Inventory (TCI-125). The factorial structure, the alpha coefficients, and the item-total correlations coefficients of the TEMPS A and the correlation coefficients between the dimensions of the TCI and the TEMPS-A subscales were then determined. RESULTS: A principal components analysis with a Varimax rotation found that 39 out of the 110 original items of the TEMPS A loaded on five factors that were interpreted as representing the cyclothymic, depressive, irritable, hyperthymic, and anxious factors. Coefficients alpha for internal consistency were 0.91 (cyclothymic), 0.81 (depressive), 0.77 (irritable), 0.76 (hyperthymic), and 0.67 (anxious) subscales. We found statistically significant positive correlations between all-but the hyperthymic subscales and harm avoidance. Positive correlations with the hyperthymic and cyclothymic, and novelty seeking and negative correlations with the remaining subscales were also recorded. Other major findings included positive correlations between the hyperthymic and reward dependence, persistence and self-directedness; positive correlation between the self-transcendence and the cyclothymic, hyperthymic and the anxious; and negative correlations between the depressive, cyclothymic, irritable, anxious and cooperativeness. LIMITATION: As the full scale anxious temperament was added after the four scales of the TEMPS-A were developed, it has only been evaluated in 345 subjects. CONCLUSIONS: These data indicate that the TEMPS-A in its shortened version is a psychometrically valid scale with good internal consistency. The proposed five subscale structure is upheld. Concurrent validity against the TCI is shown. Most importantly, for each of the temperaments, we were able to show positive attributes which are meaningful in an evolutionary context, along with traits which make a person vulnerable to mood shifts. This hypothesized dual nature of temperament, which is upheld by our data, is a desirable characteristic for a putative behavioral endophenotype in an oligogenic model of inheritance for bipolar disorder. PMID- 15780676 TI - Studies on a German (Munster) version of the temperament auto-questionnaire TEMPS A: construction and validation of the briefTEMPS-M. AB - BACKGROUND: Based on classic German concepts of a continuum between depressive, hyperthymic, cyclothymic, and irritable temperaments and affective disorder (and adding an anxious type to the four), Akiskal and co-workers developed the Temperament Evaluation of Memphis, Pisa, Paris and San Diego both in interview (TEMPS-I) and auto-questionnaire (TEMPS-A) versions. It is the aim of the present analyses to validate a brief German version of TEMPS-A. METHODS: A total of 1056 students of the Westfalische-Wilhelms-Universitat in Munster, Germany, filled out the long 110-item version of the TEMPS-A (Munster translation by Erfurth: TEMPS M) modified into a five gradation Likert format and with the items randomized. Based on this data we constructed a brief version of the TEMPS-M. In a second study, a sample of 151 students were recruited who filled out the briefTEMPS-M twice, approximately 1 month apart. RESULTS: Our psychometric procedures resulted in the retention of 35 items from the original 110. The proposed five-factor structure of the original TEMPS-A was upheld, with relatively few item reclassification (mainly due to some overlap between depressive and anxious traits). Internal consistency (Cronbach alpha values ranging from 0.69 to 0.84) and test-retest reliability were shown. Most importantly, all temperaments in the briefTEMPS-M correlated quite well (Pearson r values ranging from 0.49 to 0.72) with their respective original versions in the longer TEMPS-M. As for construct validity, significant correlation was shown with the Beck Depression Inventory for all but the hyperthymic temperament; the hyperthymic, cyclothymic and irritable correlated highest with the self-report Manic Inventory. LIMITATIONS: The study sample of university students was selective. CONCLUSIONS: We were able to construct a brief German version of the TEMPS-A auto-questionnaire. We submit this shorter version will be suitable for both clinical (psychiatric and general medical) and neurobiological research, as well as in studies on temperament features in selected populations, e.g., allowing comparisons between regions or different (German-speaking) countries. PMID- 15780677 TI - Distribution and gender effects of the subscales of a German version of the temperament autoquestionnaire briefTEMPS-M in a university student population. AB - BACKGROUND: This paper examines the distribution of the temperamental characteristics and gender effects of a new autoquestionnaire developed by Akiskal et al. (TEMPS-A) in its German briefTEMPS-M version. METHODS: As described in a companion article [J. Affect. Disord. 85 (2005), 53, this issue], based on a study population of 1056 students of the Westfalische-Wilhelms Universitat in Munster, Germany, we constructed the briefTEMPS-M. In the present paper we report on the basic descriptive statistics of the five subscales of the briefTEMPS (depressive, cyclothymic, hyperthymic, irritable, and anxious), as well as gender differences. RESULTS: Except for the hyperthymic, these subscales are capable of representing the full range of temperament in a sample of German students. Characteristics of the distribution (skewness, kurtosis) of the subscales are well in acceptable limits. We found higher depressive, cyclothymic, and anxious, as well as lower hyperthymic, temperament values in women as compared to men. Cut-off scores to determine extreme groups are provided. To render our results comparable to a similar study using the interview version of the TEMPS-I in an Italian student population [J. Affect. Disord. 47 (1998) 1; J. Affect. Disord. 51 (1998) 7], we computed the rates for dominant temperaments based on the z scores +2 S.D., and obtained the following: depressive, 4.7%; cyclothymic, 4.7%; hyperthymic, 2.1%; irritable, 4.0%; and anxious, 4.2%. CONCLUSIONS: The briefTEMPS-M is a potentially valuable scale to quickly assess temperament in research, clinical and normal samples. PMID- 15780678 TI - Factorial structure and internal consistency of the German TEMPS-A scale: validation against the NEO-FFI questionnaire. AB - BACKGROUND: The purpose of the study was to evaluate the psychometric properties of the German version of the TEMPS-A questionnaire. Besides the reliability of the temperament scales, validity was the focus of interest. Therefore, the relationship between the TEMPS-A and the well-established personality questionnaire NEO-FFI, whose factors show theoretical overlap with temperaments, was investigated. METHOD: A total of 227 students aged between 20 and 42 years were asked to fill in both instruments. Reliability coefficients for the five temperament scales and correlations among the scales of both questionnaires were calculated, as well as multiple linear regression analysis with the five personality factors and gender as independent, and the five temperaments as dependent variables. RESULTS: Reliability indices for the five temperament scales were satisfactory, with values ranging between 0.63 (depressive) and 0.76 (anxious). Women scored higher on depressive and anxious scales, whereas men had higher scores on hyperthymic temperament. Correlations within the temperament scales showed close relationships between depressive, anxious and cyclothymic temperaments; cyclothymic and irritable temperament were also related. The personality factors of the NEO-FFI predicted temperaments fairly well and explained between 41% and 58% of the variance; the main effects were exerted by neuroticism and extraversion, while the irritable temperament was primarily explained by low agreeableness. LIMITATION: The study sample was relatively small and selected. CONCLUSION: The TEMPS-A scale has sufficient reliability and good validity in a non-clinical sample. It opens new possibilities for clinical research at the interface of mental disorders, temperament and personality. Such research is in progress. PMID- 15780679 TI - Reliability and validity of TEMPS-A in a Japanese non-clinical population: application to unipolar and bipolar depressives. AB - BACKGROUND: In Japan, TEMPS-A has gathered much attention, because Kraepelin's concepts on "fundamental states" of mood disorder and temperaments have been widely respected. METHOD: TEMPS-A was translated into Japanese (and after the approval of the English back translation by H.S.A.), it was administered to 1391 non-clinical subjects, and 29 unipolar and 30 bipolar patients in remission. Of the non-clinical sample, 426 were readministered the instrument again in 1 month. A control group matched for gender and age was drawn from the non-clinical sample. RESULTS: Regarding test-retest reliability, Spearman's coefficients for depressive, cyclothymic, hyperthymic, irritable and anxious temperaments were 0.79, 0.84, 0.87, 0.81 and 0.87, respectively; regarding internal consistency, Cronbach's alpha coefficients were 0.69, 0.84, 0.79, 0.83 and 0.87, respectively. The unipolar and bipolar groups showed significantly higher depressive, cyclothymic and anxious temperament scores than the control group. Curiously, the bipolar group showed significantly lower hyperthymic score than the control group; irritable temperament scores showed no significant differences. Depressive, cyclothymic, irritable and anxious temperament scores showed significant correlations with each other. Between the unipolar and bipolar groups, there was little difference regarding the temperament scores. Also the inter-temperament correlations showed the same pattern in the unipolar and bipolar groups. LIMITATION: The clinically well cohort was 70% male. CONCLUSION: TEMPS-A showed a high reliability and validity (internal consistency) in a Japanese non-clinical sample. By and large, the hypothesized five temperament structure was upheld. Depressive, cyclothymic and anxious temperaments showed concurrent validity with mood disorder. Irritable temperament may represent a subtype of depressive, cyclothymic or anxious temperaments. There may be a temperamental commonality between unipolar and bipolar disorders. TEMPS-A will open new possibilities for international research on mood disorder and personality traits. PMID- 15780680 TI - The proposed factor structure of temperament and personality in Japan: combining traits from TEMPS-A and MPT. AB - BACKGROUND: In Japan, Kraepelin's descriptions on four "fundamental states" of manic depressive illness, the concepts of schizoid temperament by Kretschmer and obsessional and melancholic type temperament by Shimoda and Tellenbach have been widely accepted. This research investigates the construct validity of these temperaments through factor analysis. METHOD: TEMPS-A measured depressive, cyclothymic, hyperthymic and irritable temperaments and MPT rigidity, esoteric and isolation subscales measured, respectively, melancholic type and schizoid temperaments. Factor analysis was implemented with TEMPS-A alone and TEMPS-A and MPT combined data. RESULTS: With TEMPS-A alone analysis, Factor 1 included 1 depressive, 11 cyclothymic and 12 irritable temperament items with a factor loading higher than 0.4; Factor 2 included 1 depressive and 10 hyperthymic temperament items; and Factor 3 included 2 depressive temperament items only. With TEMPS-A and MPT combined data, Factor 1 included 3 depressive, 11 cyclothymic and 5 irritable temperament items with a factor loading higher than 0.4 (interpreted as the central cyclothymic tendency for all affective temperaments along Kretschmerian lines and accounting for 11.7% of the variance); Factor 2 included 6 hyperthymic temperament items (6.22% of variance); Factor 3 included 1 cyclothymic, 7 irritable and 1 schizoid temperament items (interpreted as the irritable temperament and accounting for 3.24% of the variance); Factor 4 included 1 depressive temperament and 5 melancholic type items (interpreted as the latter, accounting for 2.66% of the variance); Factor 5 included 5 depressive temperament items, along interpersonal sensitivity and passivity lines, and accounting for 2.31% of the variance; and Factor 6 included 4 schizoid temperament items accounting for 2.07% of the variance. LIMITATION: We did not use the Kasahara scale, which some believe to better capture the Japanese melancholic type. Sample was 70% male. CONCLUSION: These analyses confirm the factor validity of depressive, hyperthymic, cyclothymic and irritable temperaments (TEMPS-A), as well as the melancholic type and the schizoid temperament (MPT). Traits of the depressive and melancholic types emerge as rather distinct. Indeed, our results permit the delineation of an interpersonally sensitive type that "gives in to others" as the core features of the depressive temperament; this is to be contrasted with the higher functioning, perfectionistic, work-oriented melancholic type. Mood dysregulation is represented by the largest number of traits in this population. Contrary to a widely held belief that the melancholic type with its devotion to work and to others is the signature temperament in Japan, cyclothymic traits account for the largest variance in this nonclinical population. Hyperthymic temperament, melancholic type and schizoid temperaments appear largely independent of mood dysregulation. In this Japanese population, TEMPS-A may identify temperament constructs more comprehensively when implemented with melancholic type and schizoid temperament question items added to it. The proposed new Japanese Temperament and Personality (JTP) Scale has self-rated items divided into six subscales. PMID- 15780681 TI - Temperament and job stress in Japanese company employees. AB - BACKGROUND: This study aims to demonstrate the relevance of temperament to job stress. METHOD: The subjects were 848 male and 366 female Japanese company employees. Temperament Evaluation of Memphis, Pisa, Paris and San Diego Autoquestionnaire version (TEMPS-A) and Munich Personality Test (MPT) were administered to assess temperaments, and the NIOSH Generic Job Stress Questionnaire (GJSQ) to assess job stress. We used hierarchical multiple linear regression analysis in order to demonstrate whether temperament variables added any unique variance after controlling the effects of other predictors such as gender, age and job rank. RESULTS: In all subscales of the GJSQ, temperament predicted a large share of the variance in job stress. Remarkably, for interpersonal relationship stressors, the temperament variables added greater variance than that predicted by gender, age and job rank. Summary of the hierarchical linear regression analysis showed that the irritable temperament was associated with the most prominent vulnerability, followed by cyclothymic and anxious temperaments. The schizoid temperament had difficulty in the area of social support. On the other hand, the hyperthymic temperament displayed significant robustness in facing most job stressors; the melancholic type showed a similar pattern to a lesser degree. LIMITATION: The findings may be different in a clinical Japanese sample, or a cohort of healthy employees from a different cultural background. CONCLUSIONS: Temperament influences job stress significantly indeed, it impacts on such stress with greater magnitude than age, gender and job rank in most areas examined. Temperament influences interpersonal relationship stressors more than workload-related stressors. Interestingly, in line with previous clinical and theoretical formulations, the hyperthymic and melancholic types actually appear to be "hyper-adapted" to the workplace. PMID- 15780682 TI - Affective temperaments in clinically-well subjects in Turkey: initial psychometric data on the TEMPS-A. AB - BACKGROUND: This is a first attempt to evaluate the reliability and factor structure replicability of the Temperament Evaluation of Memphis, Pisa, Paris and San Diego Autoquestionnaire (TEMPS-A) in its Turkish Version. The questionnaire is a self-report 110-item measure that postulates five affective temperaments-the depressive, cyclothymic, irritable, hyperthymic, and anxious-which embody both strengths and liabilities along affective lines. METHODS: The questionnaire was administered to 658 clinically-well subjects in a Turkish university circle. We undertook item analysis and test-retest reliability. We then examined internal consistency through factor analysis with PCA rotation. RESULTS: We found good to excellent test-retest reliability (0.73-0.91), and internal consistency (0.77 0.85). We deleted 10 items with factor loading <0.20 for their own subscales, resulting in a questionnaire with 99 items. Despite considerable overlap between depressive and cognitive anxiety traits, a distinct "nervous"-anxious factor emerged as well, and the hypothesized (original English) 5-factor structure of the TEMPS-A was supported. Cut-offs for each temperament were based on z-scores higher than +2S.D. Dominant irritable (3.7%), nervous-anxious (3.7%) and depressive (3.1%) temperaments were the most common in this population, whereas dominant cyclothymic (1.7%) and hyperthymic (1.2%) temperaments were relatively uncommon. These temperaments tended to lose their intensity with age. As expected, women scored significantly higher on the nervous-anxious, and men on the hyperthymic temperaments. LIMITATIONS: The sample was composed of younger subjects with higher education than the general population of Turkey. Although the distribution of the scores for each of the temperaments deviated somewhat from normal curves, for heuristic reasons we did attempt to provide prevalence rates based on z-scores. CONCLUSION: In this preliminary version of the TEMPS-A, we have retained 100 (of the original 110) traits loading >0.20. Some deleted items referred to sleep, others appeared socially desirability traits in the Turkish culture endorsed by many subjects. Nonetheless, item analyses within each factor revealed traits indicative of personal assets (specific to each temperament) along with those which might represent vulnerability to affective illness. This is in line with the hypothesized original theoretical framework of the senior authors. Even in this "first pass," in its Turkish version the TEMPS-A is a reliable and valid instrument. Further refinement of the instrument will require the study of a nationally representative sample in Turkey. PMID- 15780683 TI - Affective temperaments as measured by TEMPS-A in patients with bipolar I disorder and their first-degree relatives: a controlled study. AB - BACKGROUND: The aims of this study were to identify the dominant affective temperamental characteristics of patients with bipolar disorder (BP) and their clinically well first-degree relatives and to compare the prevalence rates of these temperaments with those in healthy control subjects. METHODS: One hundred bipolar I probands and their 219 unaffected first-degree relatives were enrolled in the study. The control group consisted of healthy subjects without any personal or family history of bipolar disorder, matched with the age and gender of the probands and first-degree relatives. To identify the dominant affective temperaments, the Turkish version of TEMPS-A scale was used. RESULTS: At least one dominant temperament was found in 26% of the proband group, in 21.9% of the relative group, and 6.0% and 10.0% of the control groups, respectively. The most noteworthy finding was that both the probands and their relatives had significantly higher frequency of hyperthymic temperament than the controls. LIMITATIONS: Temperament had not been assessed premorbidly in the probands with bipolar disorder. CONCLUSIONS: The study supports the familial, possibly genetic, basis for the hyperthymic temperament in the genesis of bipolar I dosorder. That the cyclothymic temperament was not similarly represented, may be due to the higher specificity of the cyclothymic temperament to the bipolar II sybtype (which we did not study). More research is needed on the relevance of cyclothymic and other temperaments to the genetics of bipolar disorders selected by rigorous subtyping along the clinical spectrum of bipolarity. PMID- 15780684 TI - The cyclothymic temperament in healthy controls and familially at risk individuals for mood disorder: endophenotype for genetic studies? AB - BACKGROUND: The modern concept of affective disorders focuses increasingly on the study of subthreshold conditions on the border of manic or depressive episodes. Indeed, a spectrum of affective conditions spanning from temperament to clinical episodes has been proposed by the senior author. As bipolar disorder is a familial illness, an examination of cyclothymic temperament (CT) in controls and relatives of bipolar patients is of major relevance. METHODS: We recruited a total sample of 177 healthy symptom-free volunteers. These controls were divided into three groups. The first one is comprised of 100 normal subjects with a negative familial affective history (NFH); the second of 37 individuals, with positive affective family history (PFH); and a third of 40 subjects, with at least one sib or first-degree kin with bipolar disorder type I according to the DSM-IV (BPR). The last two groups defined at risk individuals. We interviewed all subjects with CT, as described by the senior author. RESULTS: We found a statistically significant difference in the rates of CT between the subjects in BPR versus others. CT was also more prevalent in the PFH compared with NFH. Additionally, the simple numeration of the CT traits exhibited gradation in the distribution of individuals inside the NFH, PFH and BPR. Finally, categorically defined CT and CT traits predominated in females. LIMITATION and CONCLUSION: Although not all relatives of bipolar probands were studied, our results exhibit an aggregation of CT in families with affective disorder-and more specifically those with bipolar background. These results allow us to propose the importance of including CT for phenotypic characterization of bipolar disorder. Furthermore, our results support a spectrum concept of bipolar disorder, whereby CT is distributed in ascending order in the well-relatives of those with depressive and bipolar disorders. We submit that this temperament represents a behavioral endophenotype, serving as a link between molecular and behavioral genetics. PMID- 15780686 TI - Familiality of temperament in bipolar disorder: support for a genetic spectrum. AB - BACKGROUND: The array of different diagnoses and clinical presentations seen in the family members of bipolar probands suggests a quantitative or spectrum phenotype. Consistent with this idea, it has been proposed that an underlying quantitative variation in temperament may be the primary phenotype that is genetically transmitted and that it in turn predisposes to bipolar disorder (BP). Choosing the appropriate phenotypic model for BP is crucial for success in genetic mapping studies. To test this theory, various measures of temperament were examined in the family members of bipolar probands. We predicted that a gradient of scores would be observed from those with BP to those with major depression to unaffected relatives to controls. METHODS: Members of 85 bipolar families and 63 control subjects were administered clinical interviews for diagnosis (SCID) and two temperament assessments, the TEMPS-A and TCI-125. Subjects with BP, major depressive disorder, unaffected relatives, and controls were compared on each temperament scale and on eight factors extracted from a joint factor analysis of the TEMPS-A and TCI-125. RESULTS: The four groups were found to be significantly different and with the expected order of average group scores for four of the TEMPS-A scales, three of the TCI-125 scales, and one of the extracted factors. On the fifth TEMPS-A scale, hyperthymic, controls scored higher than the other three subject groups contrary to expectations. Significant differences were seen between unaffected relatives and controls on the hyperthymic scale and on the first extracted factor, anxious/reactive. LIMITATIONS: Controls were mainly recruited through advertisements, which may have introduced an ascertainment bias. It is also possible that mood state at the time of completing the questionnaire influenced subject's rating of their temperament. Additionally, bipolar I and bipolar II subjects were placed in the same group even though they had some differing clinical features. CONCLUSIONS: Our data support the theory that some dimensions of temperament are transmitted in families as quantitative traits that are part of a broader bipolar spectrum. In particular, the hyperthymic scale of the TEMPS-A and the anxious/reactive extracted factor distinguished unaffected relatives from controls. The hyperthymic scale yielded results opposite to expectation with controls higher than any family group. This may be an artifact of the self-rated form of the questionnaire, a consequence of our grouping bipolar I and II subjects together, or the result of a "protective" factor and bears further study. Nevertheless, both of these scales may be useful quantitative traits for genetic mapping studies. PMID- 15780687 TI - Affective temperament traits measured by TEMPS-I and emotional-behavioral problems in clinically-well children, adolescents, and young adults. AB - OBJECTIVE: The identification of specific temperament dimensions as correlates or risk factors for psychopathology in infancy, childhood and adolescence might provide key information to elucidate causal mechanisms that underlie these relationships. METHODS: A non-clinical sample of 1010 students (518 males and 492 females) without major psychiatric disorders was given psychometric assessment using TEMPS-I (the Italian Semi-structured Interview version of the Temperament Evaluation of Memphis, Pisa, and San Diego) and EBC (Emotional and Behavioral Checklist in Infancy, Childhood and Adolescence). Grouping the subjects on the basis of the highest z-score obtained on each of the four temperament scales of TEMPS-I, it was possible to identify the dominant affective temperamental (AT) inclination of each individual: 283 (28.0%) subjects were classified as dominant depressive temperament, 446 (44.2%) as dominant hyperthymic, 221 (21.9%) as dominant cyclothymic, and 60 (5.9%) as dominant irritable. The effects of AT dominant groups on EBC scores were tested by one-way analysis of variance. To control for age and sex effects, we tested the differences within dominant AT groups by a multiple classification analysis (MCA). RESULTS: As expected, subjects with depressive temperament traits were characterized by social inhibition and lack of antisocial and hyperactive behavior. Cyclothymic subjects reported the highest number of emotional and behavioral problems, compared with the other dominant ATs (depressive, hyperthymic and irritable). In particular, a cyclothymic disposition was most frequently associated with anxiety-sleep disturbances, sensitivity to separation, eating disturbances in females and antisocial-aggressive behavior in males. The relationship between cyclothymic temperament and anxiety-sleep disturbances and antisocial-aggressive behavior increased with age. LIMITATIONS: Cross-sectional study based on retrospective evaluation. CONCLUSIONS: Within a juvenile population, depressive temperament is a construct partially overlapping with behavioral inhibition, while extremes of emotionality and behaviors occur preponderantly in those with cyclothymic traits. The cyclothymic disposition turned out to be the most 'morbid', and associated with both internalizing and externalizing disturbances. PMID- 15780685 TI - A comparison of recovered bipolar patients, healthy relatives of bipolar probands, and normal controls using the short TEMPS-A. AB - OBJECTIVE: To investigate the presence of temperament dysregulation in healthy relatives of bipolar probands (RBP), a population at high risk for developing mood disorders, by comparing them with clinically recovered bipolar patients (BP) and normal controls (NC). METHOD: 52 RBP and 23 BP were originally recruited for a multicenter genetic study in bipolar disorders. NC (n=102) were also recruited by newspaper advertisement, radio and television announcements, flyers, newsletters, or word of mouth. All volunteers were asked to complete the TEMPS-A Scale, a self-report questionnaire designed to measure temperamental variations in psychiatric patients and healthy volunteers. In scoring temperaments, we relied upon the short validated version of the TEMPS-A [J. Affect. Disord. (2004)], from which traits with loadings <0.035 had been deleted. RESULTS: To examine differences in temperament dimensions among the three groups, a MANCOVA model was constructed using diagnostic group as the fixed factor (BP vs. RBP vs. NC); effects of age and gender were adjusted as covariates. MANCOVA showed overall group effect on the dependent variables (Hotelling's F5,175=6.64, p<0.001). Four dependent variables (dysthymic, cyclothymic, irritable, and anxious temperaments) showed significant between-group differences. RBP showed lower cyclothymic temperament scores than BP, but higher scores than NC. BP and RBP showed higher anxious temperament scores than NC. Hyperthymic scores were significantly highest in the NC. LIMITATION: In view of the small cell sizes, bipolar I vs. bipolar II subanalyses could not be conducted. CONCLUSIONS: Methodologic strengths of the present analyses is that the BP group had clinically recovered, and we used the validated short version of the TEMPS-A for the present analyses. Our findings suggest that some clinically healthy relatives of bipolar probands exhibit a subclinical cyclothymic instability in mood, interest, self-confidence, sleep, and/or energy as well as anxiety proneness that is not observed among normal controls. These traits may represent vulnerability markers and could presumably be used to identify individuals at high risk for developing bipolar spectrum disorders, or specific clinical subtypes (e.g., bipolar I, bipolar II) within this spectrum. This is a conceptual perspective with many unanswered questions. Resolution of these questions will require innovative definitions of phenotypes to be included in the analyses of the temperament subscales in different populations. The temperament subscales themselves need to be calibrated properly, to find out which traits or specific combinations of trains are most promising. More extensive and complex quantitative trait analyses of these temperaments in a much expanded sample are reported elsewhere in this issue [J. Affect. Disord. (2004)]. PMID- 15780688 TI - Cyclothymic temperament as a prospective predictor of bipolarity and suicidality in children and adolescents with major depressive disorder. AB - INTRODUCTION: Although several recent studies suggest that bipolar disorder most commonly begins during childhood or adolescence, the illness still remains under recognized and under-diagnosed in this age group. As part of the French Bipolar network and in line with the hypothesis that juvenile depression is pre-bipolar , we evaluated the rate of onset of bipolar disorders in a naturalistic 2-year prospective study of consecutive, clinically depressed children and adolescents, and to test whether the cyclothymic temperament underlies such onset. METHODS: Complete information was obtained from both parents and patients in 80 of 109 depressed children and adolescents assessed with Kiddie-SADS semi-structured interview, according to DSM IV criteria. They were also assessed with a new questionnaire on cyclothymic-hypersensitive temperament (CHT) from the TEMPS-A cyclothymic scale adapted for children (provided in ), and other assessment tools including the Child Depression Inventory (CDI), Young Mania Rating Scale, Clinical Global Assessment Scale (CGAS), and Overt Aggressive Scale (OAS). RESULTS: Of the 80 subjects, 35 (43%) could be diagnosed as bipolar at the end of the prospective follow-up. This outcome was significantly more common in those with cyclothymic temperament measured at baseline. Most of these patients were suffering from a special form of bipolar disorder, characterized by rapid mood shifts with associated conduct disorders (CD), aggressiveness, psychotic symptoms and suicidality. LIMITATION: The primary investigator, who took care of the patients clinically, was not blind to the clinical and psychometric data collected. Since all information was collected in a systematic fashion, the likelihood of biasing the results was minimal. CONCLUSION: We submit that the CHT in depressed children and adolescents heralds bipolar transformation. Unlike hypomanic or manic symptoms, which are often difficult to establish in young patients examined in cross-section or by history, cyclothymic traits are detectable in childhood. Our data underscore the need for greater effort to standardize the diagnosis and treatment of pre-bipolar depressions in juvenile patients. PMID- 15780689 TI - Temperament and risky behaviors: a pathway to HIV? AB - BACKGROUND: Certain sexual behaviors and abuse of substances contribute to the risk of becoming HIV-infected. Because dispositional characteristics are likely to influence this process, we examined the association between temperament and HIV serostatus in subjects with heavy use of drugs and various sexual risk behaviors. METHODS: Two hundred and thirty individuals (81% of whom were HIV infected) were assessed for underlying temperamental characteristics, substance use, and risky sexual behaviors. Toward this goal, a factor analysis was conducted on a new self-rated Temperament Autoquestionnaire (Temperament Evaluation of Memphis, Pisa, Paris and San Diego-Autoquestionnaire (TEMPS-A)). MANOVA was used to examine overall associations between risky behaviors and temperament. Additionally, directed MANOVAs were conducted for each of six drugs of abuse (alcohol, marijuana, cocaine, opioids, other stimulants, and sedative hypnotics). RESULTS: Factor analysis confirmed the hypothesized temperament structure of five distinct factors: Cyclothymic, hyperthymic, anxious, irritable explosive, and depressive. Heavy users of cocaine, other stimulants, and alcohol were more likely to have an irritable-explosive temperament. Interestingly, heavy users of opioids were more likely to show depressive and hyperthymic traits. The inclusion of HIV status as a predictor of temperament did not alter the results. Unlike a previous study, no direct significant relationships were found between risky sexual behaviors and specific temperaments. LIMITATIONS: Relatively small sample size for a factor analytic study, and cross-sectional design, which cannot determine a cause and effect relationship. CONCLUSIONS: We succeeded in studying and validating five distinct affective temperaments in a substance abuse population. Irritable-explosive, and to a lesser extent, depressive and hyperthymic traits, were associated with substance use, and may place individuals at risk for HIV infection via that mechanism. That current risky sexual practices were not directly linked to temperament may reflect modification in sexual behavior following the acquisition of HIV disease. PMID- 15780690 TI - Temperament profiles in physicians, lawyers, managers, industrialists, architects, journalists, and artists: a study in psychiatric outpatients. AB - BACKGROUND: With the possible exception of cyclothymia in artists, there is a paucity of data in the literature on the temperament in different professions. METHODS: For this exploratory study, we sought to generate preliminary data on temperaments among psychiatric outpatients, including physicians (n=41), lawyers (n=30), managers and executives (n=35), industrialists (n=48), architects (n=27), journalists (n=34), and a mixed group of artists (n=48). They were compared with age, sex, social class, and affective disorder matched outpatients outside of these professions, drawn from the same clinical settings to serve as our Comparison Group (CG, n=120). We used an interview version of the Akiskal-Mallya criteria for temperaments. We finally used the DSM-III-R obsessive compulsive personality (OC traits). RESULTS: Compared with the CG, lawyers and physicians had high rates of dysthymic temperament and OC traits. Managers, like lawyers and doctors, had high rates on OC traits but were different in being very low on cyclothymic and twice as hyperthymic than the CG was. Industrialists, who, by definition, were self-made, had even higher rates of hyperthymic traits. Both architects and artists seemed to have benefited from being cyclothymic (3-4 times higher than CG's); interestingly, architects had higher levels of OC traits, and artists were less obsessional than the CG was. Overall, among managers/executives and lawyers, 41% met criteria for affective temperaments, whereas the equivalent rate among the remainder was 77%. LIMITATION: Given that this is a chart review of existing clinical records, it was not possible to be blind to the profession of the patients. A mixed group of artists may have obscured differences among artists from different domains of art (e.g., poets vs. performing artists), and the same can be said of physicians (e.g., internists vs. surgeons). A disclaimer would be appropriate: Ours is not a study on eminence in the different professions but on the temperament and personality profiles that distinguish among them. CONCLUSIONS: Despite the foregoing limitations and overlapping attributes in the different professions, they nonetheless emerged as having distinct temperamental and personality profiles. Dysthymic and obsessional attributes are notable in lawyers and physicians. We confirm the role of cyclothymia in artists and architects. The role of the hyperthymic temperament in managers, self-made industrialists, and journalists, to the best of our knowledge, is being reported for the first time. The role of cyclothymic and hyperthymic temperaments appears to be moderated by obsessional traits across the entire professional realm examined. In particular, artists' creative imagination appears "liberated" by low levels of OC traits, whereas among architects, relatively high levels of OC traits seem to contribute to the execution of their work. More tentatively, judging from the overall levels of affective temperaments in the remaining professions, on average, more of the managers/executives than self-made industrialists could be described as "colder" in temperament, and more of the physicians "warmer" than lawyers are. Journalists, as a group, appeared to possess the broadest representation of affective temperaments. The foregoing conclusions must be regarded as tentative, even hypothetical, in need of verification among professionals without major psychiatric disorders. Nonetheless, temperament profiles among psychiatrically ill professionals in the seven professional realms studies can help predict how they relate to their doctors, family members, colleagues, coworkers, and clients/patients. Such knowledge, in turn, can help the therapeutic process. PMID- 15780691 TI - Temperamental commonalities and differences in euthymic mood disorder patients, creative controls, and healthy controls. AB - OBJECTIVE: Understanding of mood disorders can be enhanced through assessment of temperamental traits. We explored temperamental commonalities and differences among euthymic bipolar (BP) and unipolar (MDD) mood disorder patients, creative discipline graduate student controls (CC), and healthy controls (HC). METHODS: Forty-nine BP, 25 MDD, 32 CC, and 47 HC completed self-report temperament/personality measures including: The Affective Temperament Evaluation of Memphis, Pisa, Paris and San Diego (TEMPS-A); the Revised NEO Personality Inventory (NEO-PI-R); and the Temperament and Character Inventory (TCI). RESULTS: Euthymic BP, MDD, and CC, compared to HC, had significantly increased cyclothymia, dysthymia and irritability scores on TEMPS-A; increased neuroticism and decreased conscientiousness on NEO-PI-R; and increased harm avoidance and novelty seeking as well as decreased self-directedness on TCI. TEMPS-A cyclothymia scores were significantly higher in BP than in MDD. NEO-PI-R openness was increased in BP and CC, compared to HC, and in CC compared to MDD. TCI self transcendence scores in BP were significantly higher than in MDD, CC, and HC. LIMITATIONS: Most of the subjects were not professional artists, and represented many fields; temperament might be different in different art fields. CONCLUSIONS: Euthymic BP, MDD, and CC compared to HC, had prominent temperamental commonalities. However, BP and CC had the additional commonality of increased openness compared to HC. BP had particularly high Cyclothymia scores that were significantly higher then those of MDD. The prominent BP-CC overlap suggests underlying neurobiological commonalities between people with mood disorders and individuals involved in creative disciplines, consistent with the notion of a temperamental contribution to enhanced creativity in individuals with bipolar disorders. PMID- 15780692 TI - Proposal for a bipolar-stimulant spectrum: temperament, diagnostic validation and therapeutic outcomes with mood stabilizers. AB - BACKGROUND: Stimulant abuse and dependence in confusing admixtures with features of bipolar disorder has been variously conceptualized. DSM-IV-TR rules tend to be liberal in permitting the diagnosis of substance-induced disorder, and rather strict for a bipolar diagnosis in such cases. Accordingly, if bipolarity is ever considered in this interface, it usually pertains to syndromal bipolar I disorder. METHOD: We therefore focused on the more prevalent but relatively neglected admixture of stimulant abuse with bipolar II and lesser degrees of bipolarity. We examined in our dual diagnosis program the longitudinal progression of the dual pathology in 10 patients who had proven refractory to past efforts to treat the stimulant component. In doing so, we wished to better delineate the nature of the clinical interface of the stimulant bipolar spectrum and its response to anticonvulsant mood stabilizers. RESULTS: In nearly all cases cyclothymic and hyperthymic traits preceded the use of stimulants by years, which seemed to serve the purpose of controlling or maintaining the subthreshold rewarding mood condition. Eventually clinically more ominous and socially destructive pathology evolved, with contributions from both the bipolar diathesis and the addictive process. Seven of 10 cases had bipolar familial indicators. Nearly all evidenced hypomanic and/or irritable depressive states with mixed features during protracted sobriety from the stimulant. Except for two of the 10 patients, substantial (30-45 point) gains were made on DSM-IV-TR axis V general assessment of functioning (GAF) scores with the use of largely "mood-stabilizing" anticonvulsants. This was paralleled with the reduction of craving. These data highlight the human dimensions of the bipolar-stimulant abuse interface, and document functional outcomes (rather than mere changes in rating scales which may not necessarily reflect clinically relevant improvement). LIMITATION: Open case series of 10 patients. CONCLUSION: We propose a bipolar-stimulant spectrum-what the senior author has elsewhere labeled bipolar III-1/2-where subthreshold bipolar traits are complicated by stimulant abuse, eventually leading to pathology characteristic of both disorders. The contribution of bipolarity to this spectrum is supported by: (1) premorbid cyclothymic and hyperthymic traits; (2) familial bipolarity; (3) presence of subthreshold bipolar signs and symptoms during protracted sobriety. We further submit that anticonvulsants in this spectrum not only treat the acute escalation of activated and mixed depressive states, withdrawal phenomena, and craving for the stimulant, but also the craving for activation and mood enhancement of the underlying temperament. We submit that the latter might be crucial for the successful attenuation of the underlying diathesis for stimulant seeking behavior, abuse and dependence. PMID- 15780693 TI - The theoretical underpinnings of affective temperaments: implications for evolutionary foundations of bipolar disorder and human nature. AB - We sketch out putative evolutionary roles for affective temperaments within the theoretical framework of mood disorders conceptualized as extremes in an oligogenic model of inheritance, whereby the constituent traits in their dilute phenotypes confer adaptive advantages to individuals and/or their social group. Depressive traits, among other functions, would subserve sensitivity to the suffering of other members of the species, overlapping with those of the generalized anxious temperament, thereby enhancing the survival of not only kin but also other conspecifics. The pursuit of romantic opportunities in cyclothymia suggests that it may have evolved as a mechanism in reproductive success; cyclothymics' creative bent in poetry, music, painting, cooking or fashion design (among men, in particular) also appears useful for sexual seduction. Hyperthymic traits would lend distinct advantages in leadership, exploration, territoriality and mating. These are just some of the possibilities of the rich and complex temperamental traits subserving bipolarity within an evolutionary framework. We test selected aspects of these hypotheses with the use of correlations between the constituent traits of the Temperament Evaluation of Memphis, Pisa, Paris and San Diego (TEMPS) and correlations between the TEMPS and the Temperament and Character Inventory (TCI). Such data support the counterbalancing protective influence of harm avoidance on the risk-taking behavior of cyclothymic individuals, in both men and women. Finally, we outline a hypothesis on the evolutionary function of anxious-depressive traits for women. PMID- 15780694 TI - Agitated "unipolar" depression re-conceptualized as a depressive mixed state: implications for the antidepressant-suicide controversy. AB - BACKGROUND: The nosologic status of agitated depression is unresolved. Are they unipolar (UP) or bipolar (BP)? Are they mixed states? Even more controversial is the notion that antidepressants might play some role in the suicidality of such patients (Akiskal and Mallya, 1987) [Akiskal, H.S., Mallya, G., 1987. Criteria for the "soft" bipolar spectrum: treatment implications. Psychopharmacol Bull. 23, 68-73]. METHODS: After excluding all patients with history of hypomanic episodes occurring outside the frame of a major depressive episode (MDE), even those with a shorter duration of hypomanic symptoms than stipulated in DSM-IV, the remaining consecutive 254 unipolar major depressive disorder (MDD) private adult (> 21 years old) outpatients were interviewed (off psychoactive drugs for 2 weeks) with the Structured Clinical Interview for DSM-IV (SCID-CV), the Hypomania Interview Guide (HIGH-C), and the Family History Screen. Intra-MDE hypomanic symptoms were systematically assessed, with > or = 3 such symptoms required for a diagnosis of depressive mixed state (DMX). Agitated depression was defined as an MDE with HIGH-C psychomotor agitation score > or = 2. Logistic regression was used to study associations and control for confounding variables. RESULTS: In this strictly defined unipolar sample, agitated depression was present in 19.7%. Compared with its non-agitated counterpart, it had significantly fewer recurrences, less chronicity, higher rate of family history for bipolar disorder, and DMX; and, among the intra-depressive non-euphoric hypomanic symptoms (in decreasing order of frequency), distractibility, racing/crowded thoughts, irritable mood, talkativeness, and risky behavior. The most striking finding was the robust association between agitated depression and DMX (OR = 36.9). Furthermore, patients with psychomotor agitation had significantly higher rate of weight loss and suicidal ideation. Of DMX symptoms, we found an association between suicidal ideation, psychomotor activation, and racing thoughts. Agitated depression was tested by forward stepwise logistic regression versus all variables significantly different in the pairwise comparisons, yielding DMX, talkativeness, and suicidal ideation as the independent significant positive predictors. LIMITATIONS: No suicidal ideation scale was used. CONCLUSIONS: Agitated depression emerges as a distinct affective syndrome with weight loss, pressure of speech, racing thoughts and suicidal ideation. Psychomotor activation and racing thoughts during MDD independently predicted suicidal ideation. In this "unipolar" MDD sample, agitated depression had a strong clustering of intra episode non-euphoric hypomanic symptoms (i.e. DMX) which, coupled with its association with bipolar family history, support its link with the bipolar spectrum. Agitated depression is therefore best regarded as "pseudo-unipolar." These findings overall accord with classical German concepts of agitated depression as a mixed state. Given that these patients are typically activated along the lines of risk-taking behavior, Kraepelin's rubric of "excited (mixed) depression" appears to us the preferred terminology over "agitated depression". CLINICAL IMPLICATIONS: The data reported herein, placed in the setting of the literature reviewed in the discussion suggest that the reports of increased risk of suicidal ideation and/or behavior in some depressed patients treated by antidepressant monotherapy or combinations thereof might be attributed to baseline psychomotor activation/agitation as part of an unrecognized bipolar mixed state. Whether antidepressants induce de novo suicidality in MDD cannot be answered without adequately powered prospective double-blind studies, unlikely to be conducted because of ethical constraints. Nonetheless, we submit that agitated, activated, or otherwise excited depressions (which we consider as depressive mixed states) overlap considerably with the so-called antidepressant "activation syndrome." Furthermore, the rare occurrence of suicidality on antidepressants should not obscure the fact that the advent of the new antidepressants is associated with worldwide decline in suicide rates. We finally wish to point out that our formal nosology (i.e. DSM-IV and ICD-10), in its failure to recognize the bipolar nature of depressive mixed states, thereby fails to shield pseudo-unipolar patients from antidepressant monotherapy, which is inappropriate for such patients. PMID- 15780695 TI - Divalproex in the treatment of bipolar depression: a placebo-controlled study. AB - BACKGROUND: The treatment of bipolar disorder in the depressed phase is complicated by a tendency for conventional antidepressant drugs to worsen the course of the illness by precipitating a manic episode or increasing cycle frequency. Thus, the potential antidepressant efficacy of mood stabilizers, such as divalproex, which is an effective treatment for the manic phase of bipolar disorder, is of considerable interest. METHODS: The clinical efficacy of divalproex (valproate, Depakote) was tested in an 8-week, double-blind, placebo controlled, randomized clinical trial in 25 outpatients with bipolar I depression. The primary outcome measure was the 17-item Hamilton Rating Scale for Depression, and secondary measures included the Hamilton Rating Scale for Anxiety, the Clinician Administered Rating Scale for Mania, and the Clinical Global Impression scale. RESULTS: Using repeated measures ANOVA with last observation carried forward, divalproex was more effective than placebo in improving symptoms of depression (p = 0.0002) and symptoms of anxiety (p = 0.0001) than placebo. LIMITATIONS: The sample size was small, and most patients were male. CONCLUSIONS: These pilot results indicate that divalproex is effective in reducing the symptoms of depression and anxiety in bipolar I, depressed phase. These positive results support the need to perform a larger, multisite study of divalproex treatment for bipolar depression. PMID- 15780696 TI - Personality of seasonal affective disorder analyzed by Tri-dimensional Personality Questionnaire. AB - BACKGROUND: Although there have been numerous reports in personality of mood disorders, there have been few reports in regard with personality of winter seasonal affective disorder (SAD). Furthermore, no reports have been published concerning summer SAD personality characteristics. Thus, this study was conducted to assess the personality of winter and summer SAD using Tri-dimensional Personality Questionnaire (TPQ) that have been used in a variety of mental disorders. METHODS: A total of 6135 Japanese were evaluated with TPQ, the Seasonal Pattern Assessment Questionnaire (SPAQ) and the Self-rating Depression Scale (SDS). Winter, summer and non-SAD groups were classified by SPAQ. We compared the difference of personality trait among these three groups in consideration of gender, age and SDS score influence. RESULTS: Winter SAD demonstrated higher "Novelty Seeking" and "Harm Avoidance"; summer SAD showed higher "Harm Avoidance" than the non-SAD group. "Harm Avoidance" in both SAD groups was re-analyzed using SDS score as a covariate, and "Novelty Seeking" in winter SAD using age as a covariate. As a result, the significance of high "Novelty Seeking" and high "Harm Avoidance" in winter SAD was excluded. However, "Harm Avoidance" remained the significant difference between summer and non-SAD. LIMITATION: SAD was diagnosed only by SPAQ and not by interview. The state dependency of "Harm Avoidance" was not confirmed in identical patients over lapse of time. CONCLUSION: Patients with winter SAD have high "Harm Avoidance" dependent on the depressive state that is in accordance with non-seasonal depression. Patients with summer SAD have high "Harm Avoidance" possibly independent from the depressive state. PMID- 15780697 TI - Pretreatment pattern of symptom expression in premenstrual dysphoric disorder. AB - BACKGROUND: Use of intermittent dosing strategies for the treatment of premenstrual dysphoric disorder (PMDD) highlights the need for detailed empirical data on the onset, duration and pattern of symptom expression in women suffering from PMDD. METHOD: Data were analyzed from 276 women who met DSM-IV criteria for PMDD and prospectively charted two menstrual cycles prior to commencing sertraline treatment. The presence and severity of PMDD symptoms were measured using the Daily Record of Severity of Problems (DRSP). RESULTS: The most frequent PMDD symptoms (moderate-to-severe for > or = 3 days) included anger/irritability (76%), anxiety/tension (71%), tired/lethargic (58%), and mood swings (58%). Mean DRSP scores peaked at day -2 (2 days prior to the onset of menses), but the within-patient day of onset of PMDD-level symptoms was highly variable, differing from cycle-to-cycle by > or = 4 days in 45% of women. Similarly, the within patient duration of PMDD symptoms varied from cycle-to-cycle by 3 or more days in > or = 50% of women. Depending on the criteria used, 1 day after the onset of menstruation, 34-46% of women continued to report moderate to severe symptoms. LIMITATION: Women in this sample were recruited for participation in a treatment study, and the results may not generalize to women with PMDD in the community. CONCLUSION: The results of this analysis found significant within-patient variability in the time-to-onset and offset of PMDD symptoms, as well as their duration. The temporal pattern and high degree of within-patient variability across menstrual cycles of PMDD symptoms may have treatment implications. PMID- 15780698 TI - Clinical evolution after self-induced or accidental traumatism: a controlled study of the extent and the specificity of suicidal catharsis. AB - BACKGROUND: The cathartic effect has been related to the short-term decrease of depressive symptomatology following the suicide attempt. This study aimed to clarify the extent of this clinical improvement to non self-induced traumas and other suicidal dimensions. METHODS: Twenty-six recent suicide attempters were compared with 21 control subjects admitted to the surgical ward after an accident induced trauma. They completed several assessments the day after the admission and one week after discharge: Hamilton and Carroll Depression scales, Barratt Impulsivity Scale, Hopelessness scale, State-Trait Anxiety Inventory, MMPI (abbreviated version), Global Assessment Scale. RESULTS: Depression, anxiety state, two sub-scales of the MMPI (Hysteria, Depression) and general functioning improved significantly in the suicide attempters group. Measures of impulsivity and hopelessness remained stable during the follow-up. LIMITATIONS: These results cannot not be generalized to all suicide attempters and we did not take account of the lethality and severity of the suicidal method used. CONCLUSION: Our study supports the specific role of deliberate self-aggression in the cathartic effect and the trait value of both impulsivity and hopelessness. PMID- 15780699 TI - Diagnostic stability in bipolar disorder in clinical practise as according to ICD 10. AB - BACKGROUND: The diagnostic stability of the ICD-10 diagnosis of mania/bipolar disorder has not been investigated in clinical practice. METHODS: All patients who got a diagnosis of mania/bipolar disorder at least once in a period from 1994 to 2002 at outpatient treatment or at discharge from psychiatric hospitalisation in Denmark were identified in a nationwide register. RESULTS: Totally, 4116 patients got a diagnosis of mania/bipolar disorder at least once; among these, 2315 patients (56.2%) got the diagnosis at the first contact, whereas the remaining patients (43.8%) got the diagnosis at later contacts. Approximately 30% of patients with an initial diagnosis of mania/bipolar disorder eventually changed diagnosis during follow-up. A substantial proportion of patients initially presented with prodromal syndromes such as transient psychosis, reaction to stress/adjustment disorder or mental and behavioural disorder due to psychoactive substance use and got a diagnosis of bipolar disorder later on. Especially younger but also female patients were at increased risk of delay of the diagnosis of bipolar disorder. LIMITATIONS: Only patients from psychiatric settings were included. CONCLUSIONS: Clinicians should, especially in younger and female patients, be more observant on manic symptoms in patients who as first glance presents with transient psychosis, reaction to stress/adjustment disorder or with psychoactive substance abuse and follow these patients more closely over time identifying putable hypomanic and manic symptoms as early as possible. PMID- 15780700 TI - Prevalence and distinct correlates of anxiety, substance, and combined comorbidity in a multi-site public sector sample with bipolar disorder. AB - BACKGROUND: Recent data indicate high prevalence of both anxiety and substance comorbidity in bipolar disorder. However, few studies have utilized public sector samples, and only one has attempted to separate contributions of each type of comorbidity. METHODS: 328 inpatient veterans with bipolar disorder across 11 sites were assessed using selected Structured Clinical Interview for DSM-IV modules and self-reports. RESULTS: Comorbidity was common (current: 57.3%; lifetime: 78.4%), with multiple current comorbidities in 29.8%. Substance comorbidity rate was comparable to rates typically reported in non-veteran inpatient samples (33.8% current, 72.3% lifetime). Selected anxiety comorbidity rates exceeded those in other inpatient samples and appeared more chronic than episodic/recurrent (38.3% current, 43.3% lifetime). 49% of PTSD was due to non combat stressors. Major correlates of current substance comorbidity alone were younger age, worse marital status, and higher current employability. Correlates of current anxiety comorbidity alone were early age of onset, greater number of prior-year depressive episodes, higher rates of disability pension receipt, and lower self-reported mental and physical function. Combined comorbidity resembled anxiety comorbidity. LIMITATIONS: This is a cross-sectional analysis of acutely hospitalized veterans. CONCLUSIONS: Distinct patterns of substance and anxiety comorbidity are striking, and may be subserved by distinct neurobiologic mechanisms. The prevalence, chronicity and functional impact of anxiety disorders indicate the need for improved recognition and treatment of this other dual diagnosis group is warranted. Clinical and research interventions should recognize these divergent comorbidity patterns and provide individualized treatment built "from the patient out." PMID- 15780701 TI - Luteal phase treatment of premenstrual dysphoric disorder improves symptoms that continue into the postmenstrual phase. AB - BACKGROUND: Despite the proven efficacy of luteal phase medication dosing for women with premenstrual dysphoric disorder (PMDD), it is not known whether this approach adequately treats symptoms that linger into the first 2-3 days of the follicular phase, a time when up to one-third of women diagnosed with PMDD report residual symptoms. Furthermore, no previous study has explored whether abruptly stopping medication after 2 weeks of treatment is associated with discontinuation symptoms. METHODS: To evaluate the efficacy of luteal phase medication dosing, symptom data from the Daily Record of Severity of Problems (DRSP) during first few days of menses were compared from two studies with similar designs but different treatment strategies. The first study used continuous dosing of sertraline, 50-150 mg/day, throughout the menstrual cycle, while the second study used intermittent dosing with sertraline, 50-100 mg/day in the 14-16 days prior to onset of menses. To investigate whether abruptly stopping pills led to discontinuation symptoms, DRSP data for the first 5 days after the onset of menses were analyzed in the second (intermittent dosing) study. Symptom scores were compared for subjects who took either sertraline or placebo premenstrually and ceased taking pills at the onset of menses. RESULTS: The baseline (pretreatment) to on-treatment effect sizes were similar for continuous vs. luteal phase dosing on the first day of menses (0.73 vs. 0.89), second day of menses (0.40 vs. 0.55), and third day of menses (0.42 vs. 0.44), respectively. Subjects who abruptly discontinued sertraline had fewer symptoms indicative of withdrawal at Day 3 (p < 0.01) and no difference during Days 4-5 compared to subjects abruptly discontinuing placebo. CONCLUSION: Patients given active medication during the luteal phase demonstrate reductions in DRSP total scores into the first few days of menses regardless of whether active treatment was continuous throughout the menstrual cycle or was discontinued at the onset of menses. This analysis finds no support for discontinuation symptoms following abrupt cessation of sertraline after 2 weeks of treatment for two cycles. PMID- 15780702 TI - Pre- and postnatal depression and coping: a comparative approach. AB - BACKGROUND: The assessment of perinatal depressions and coping style. METHODS: With depression scales (EPDS, GHQ.12) and Carver's BriefCope, the authors compared the prevalence rates of pre and postnatal depression in a cohort of 277 French women. RESULTS: Their work revealed very high levels of prenatal depression (almost 20%) and less important but nonetheless sizeable rates (11%) of postnatal depression, making such perinatal depressions a major public health concern. The coping styles proposed in Carver's public health BriefCope questionnaire make it possible to significantly differentiate during these two periods between depressive women and their non-depressed counterparts. CONCLUSIONS: This enables us to underline factors of risk and protection suggesting the importance of setting up compensatory and preventive systems and evaluating their pertinence in the framework of future research. PMID- 15780703 TI - Vascular risk and low serum B12 predict white matter lesions in patients with major depression. AB - BACKGROUND: While patients with depression have been shown to have a greater incidence of vascular risk factors and structural brain changes, any association with dietary co-factors is unclear. METHODS: Forty-seven patients with major depression (mean age = 52.8 years, SD = 12.6) and 21 healthy volunteers (mean age = 54.7 years, SD = 9.1) underwent high-resolution magnetic resonance imaging scanning. T2-weighted films were scored for deep white matter (DWM), periventricular (PV), and subcortical (SC) hyperintensities. RESULTS: There was no difference in lesion severity between patients and control subjects. After controlling for age, vitamin B12 levels were predictive of DWM lesions in patients. DWM and SC lesions were associated with histories of hypertension and diabetes. LIMITATIONS: A relatively small sample of patients were recruited from specialist services and the findings may not represent those observed in larger or community-based cohorts. CONCLUSIONS: In patients with major depression, vitamin B12 levels and histories of hypertension and/or diabetes are predictive of white matter lesions. PMID- 15780704 TI - Do venlafaxine XR and paroxetine equally influence negative and positive affect? AB - BACKGROUND: We assessed the therapeutic effects of venlafaxine XR and paroxetine on mood and anxiety symptoms derived from the tripartite model of mood. We hypothesized that the two antidepressants would have largely similar effects on symptoms of negative affect because both agents influence serotonergic systems. However, based on evidence indicating linkages between catecholaminergic activity and the emotional dimension of positive affect, we hypothesized that the catecholaminergic effects of venlafaxine XR would yield particularly pronounced effects on symptoms of positive affect. METHODS: Twenty depressed outpatients were randomly assigned to treatment with either venlafaxine XR (225 mg/day) or paroxetine (30 mg/day) during a 12-week treatment trial. Weekly mood ratings were collected using the Mood and Anxiety Symptom Questionnaire [Watson, D., Clark, L.A., Weber, K., Assenheimer, J.S., Strauss, M.E., McCormick, R.A., 1995. Testing a tripartite model: II. Exploring the symptom structure of anxiety and depression in student, adult, and patient samples. J. Abnorm. Psychol. 104 (1), 15-25] [Watson, D., Weber, K., Assenheimer, J.S., Clark, L.A., Strauss, M.E., McCormick, R.A., 1995. Testing a tripartite model: I. Evaluating the convergent and discriminant validity of anxiety and depression symptom scales. J. Abnorm. Psychol. 104 (1), 3-14]. RESULTS: Consistent with predictions, analyses revealed that there were no significant differences between venlafaxine XR and paroxetine on measures of negative affect. However, contrary to predictions, the two medications produced similar changes on measures of positive affect. LIMITATIONS: Replication and extension using a larger sample size are mandated. CONCLUSIONS: These preliminary results suggest that two antidepressants that appear to have dissimilar mechanisms of action may nevertheless have similar effects on the positive and negative affective components of depression. Alternatively, paroxetine may have a clinically relevant noradrenergic effect at the dose tested. PMID- 15780705 TI - Relationship of white matter hyperintensities to cerebrospinal fluid glucose polyol pathway metabolites-a pilot study in treatment-resistant affective disorder patients. AB - BACKGROUND: Magnetic resonance imaging (MRI) white matter hyperintensities (WMHs) are found at higher rates in patients with affective disorders, particularly late life or treatment-resistant disorders. Studies support a vascular pathogenesis for WMHs in late-life onset disorders; however, pathogenesis in typical early life onset disorders is less clear. Based on associations between diabetes mellitus and both WMHs and affective disorders, this study investigated the relationship between WMHs and brain glucose metabolism by the polyol pathway-a pathway linked to nervous tissue disease in diabetes. METHODS: Burdens of fluid attenuated inversion recovery (FLAIR) WMHs were quantified and correlated with cerebrospinal fluid (CSF) concentrations of glucose metabolites in 10 nondiabetic inpatients with treatment-resistant bipolar, unipolar, and schizoaffective disorders and 10 nondiabetic control patients who had been investigated clinically for transient neurological symptoms. RESULTS: Deep but not periventricular WMH burden correlated positively and significantly with elevated CSF concentrations of sorbitol, the specific polyol pathway metabolite of glucose (rho=0.86, p=0.002), in the affective disorders but not the control group. LIMITATIONS: This was a pilot study with a relatively small number of subjects; therefore, conclusions are tentative. Controls were not healthy subjects; they were patients with transient neurological symptoms. CONCLUSIONS: This is the first reported evidence of a relationship between WMHs and increased brain glucose metabolism by the polyol pathway in patients with affective disorders. More extensive studies are necessary to determine whether this preliminary finding represents a pathogenetic relationship. PMID- 15780706 TI - Pharmaceuticals: a threat to drinking water? AB - Recently, considerable interest has developed regarding the presence of pharmaceuticals in the environment, but there has been comparatively little study on the potential of these substances to enter potable supplies. This is surprising because drinking water would provide a direct route into the body for any drugs that might be present. Although many countries employ advanced treatments, such as granular activated carbon, membrane technologies, ozonation and ultraviolet radiation, for treating water intended for human consumption, some compounds have been shown to be unaffected by such processes. Here, we examine the levels of drug substances reported in drinking water around the world. The possible implications of the presence of these compounds are highlighted and assessed, and recommendations are made for further research. PMID- 15780707 TI - Nanobiotechnology: the promise and reality of new approaches to molecular recognition. AB - Nanobiotechnology is the convergence of engineering and molecular biology that is leading to a new class of multifunctional devices and systems for biological and chemical analysis with better sensitivity and specificity and a higher rate of recognition. Nano-objects with important analytical applications include nanotubes, nanochannels, nanoparticles, nanopores and nanocapacitors. Here, we take a critical look at the subset of recent developments in this area relevant to molecular recognition. Potential benefits of using nano-objects (nanotubes, quantum dots, nanorods and nanoprisms) and nanodevices (nanocapacitors, nanopores and nanocantilevers) leading to an expanded range of label multiplexing are described along with potential applications in future diagnostics. We also speculate on further pathways in nanotechnology development and the emergence of order in this somewhat chaotic, yet promising, new field. PMID- 15780708 TI - Environmental microbiology-on-a-chip and its future impacts. AB - Rapid advances in microfabrication, DNA and protein microarray and microfluidic technologies have enabled the development of fully-integrated, miniaturized systems. These so called 'laboratory-on-a-chip' (LOC) devices perform sample preparation (i.e. concentration, separation and purification) together with biochemical reactions and detection steps in a simple and automated manner. We believe LOC technology for environmental microbiology studies will have immediate impacts on microbial monitoring by achieving detection and identification within minutes at the single-cell level, and on microbial ecology by deepening the understanding of microbial community structure and diversity and correlating these with niche-specific functions within a micro space. In the long run, significant impacts are anticipated on environmental metagenomics and proteomics. PMID- 15780709 TI - Bringing medicinal plants into cultivation: opportunities and challenges for biotechnology. AB - Consumption of herbal medicines is widespread and increasing. Harvesting from the wild, the main source of raw material, is causing loss of genetic diversity and habitat destruction. Domestic cultivation is a viable alternative and offers the opportunity to overcome the problems that are inherent in herbal extracts: misidentification, genetic and phenotypic variability, extract variability and instability, toxic components and contaminants. The use of controlled environments can overcome cultivation difficulties and could be a means to manipulate phenotypic variation in bioactive compounds and toxins. Conventional plant-breeding methods can improve both agronomic and medicinal traits, and molecular marker assisted selection will be used increasingly. There has been significant progress in the use of tissue culture and genetic transformation to alter pathways for the biosynthesis of target metabolites. Obstacles to bringing medicinal plants into successful commercial cultivation include the difficulty of predicting which extracts will remain marketable and the likely market preference for what is seen as naturally sourced extracts. PMID- 15780710 TI - Biosensors based on binding-modulated donor-acceptor distances. AB - The promising recognition characteristics exhibited by biomolecules have caused significant interest in biomolecule-based sensor strategies. Here we review several emerging biosensor designs that use modulated electron or energy transfer to a bio-specific ligand as the signaling mechanism. The efficiencies of both electron transfer and energy transfer are strongly dependent on donor-acceptor distance. When coupled with the large conformational changes sometimes associated with biomolecular recognition, these distance-dependent processes provide a robust means for generating optical and electronic signals. PMID- 15780711 TI - Biocatalytic preparation of natural flavours and fragrances. AB - During the past years biocatalytic production of fine chemicals has been expanding rapidly. Flavours and fragrances belong to many different structural classes and therefore represent a challenging target for academic and industrial research. Here, we present a condensed overview of the potential offered by biocatalysis for the synthesis of natural and natural-identical odorants, highlighting relevant biotransformations using microorganisms and isolated enzymes. The industrial processes based on biocatalytic methods are discussed in terms of their advantages over classical chemical synthesis and extraction from natural sources. Recent applications of the biocatalytic approach to the preparation of the most important fine odorants are comprehensively covered. PMID- 15780712 TI - Seamless cloning and gene fusion. AB - Gene fusion technology is a key tool in facilitating gene function studies. Hybrid molecules in which all the components are joined precisely, without the presence of intervening and unwanted extraneous sequences, enable accurate studies of molecules and the characterization of individual components. This article reviews situations in which seamlessly fused genes and proteins are required or desired and describes molecular approaches that are available for generating these hybrid molecules. PMID- 15780713 TI - Immuno-PCR: high sensitivity detection of proteins by nucleic acid amplification. AB - Nucleic acid amplification techniques are used for signal generation in antibody based immunoassays, thereby dramatically enhancing the sensitivity of conventional immunoassays. Methodological aspects, as well as applications of this novel approach, are summarized in this review, with an emphasis on immuno polymerase chain reaction (IPCR). IPCR is based on chimeric conjugates of specific antibodies and nucleic acid molecules, the latter of which are used as markers to be amplified by PCR for signal generation. The enormous efficiency of nucleic acid amplification typically leads to a 100-10,000-fold increase in sensitivity, as compared with the analogous enzyme-amplified immunoassay. The evolution of IPCR included the development of efficient reagents, the design of assay formats and the maintenance of functionality, even within complex biological matrices. Eventually, IPCR crossed the border from being a research method to a routine laboratory technique, enabling a broad range of applications in immunological research and clinical diagnostics. PMID- 15780714 TI - Prospective population-based incidence of Haemophilus influenzae type b meningitis in Thailand. PMID- 15780715 TI - Vaccines for otitis media: proposals for overcoming obstacles to progress. AB - Otitis media is a common problem with enormous morbidity worldwide. The development of vaccines to prevent otitis media would have an important human and economic impact. A striking lack of progress in the development, production and clinical testing of vaccines to prevent otitis media has occurred in the past decade. This review outlines a series of specific proposals intended to advance vaccine development for otitis media. PMID- 15780716 TI - Mucosal targeting of allergen-loaded microspheres by Aleuria aurantia lectin. AB - Murine intestinal M-cells express alpha-L-fucose residues. We constructed alpha-L fucose-targeting particles for oral immunotherapy of IgE-mediated allergy. Poly(D,L-lactic-co-glycolic acid)-microspheres were loaded with birch pollen allergens, and functionalised with the alpha-L-fucose specific Aleuria aurantia lectin (AAL). The AAL-microspheres had a size of 1-3 microm, protected the entrapped allergens from gastric degradation and released 46.6+/-1.3% allergen over 21 days in vitro. Oral gavages of AAL-particles to naive BALB/c mice induced birch pollen-specific IgG2a, but not IgG1 antibodies. We conclude that targeting allergens to alpha-L-fucose-receptor bearing cells using AAL-microspheres induces specific Th1-antibody responses possibly counteracting Th2-dominated allergy, and therefore provides a potentially useful formulation for oral immunotherapy. PMID- 15780717 TI - Mumps in Poland since 1990 to 2003; epidemiology and antibody prevalence. AB - The annual incidence of mumps in Poland over the period 1990-2003 was 45-570 per 100,000 population with the epidemic peak every 4-5 years. Till 2003, mumps vaccination has not been included in the part of National Immunisation Program in Poland that comprises the obligatory vaccinations. However, mumps vaccination was recommended by National Health Authority for children at the second year of life and it could be obtained privately. The proportion of vaccinated children has increased by 50% in last years. It has influenced on decreasing of number of notified mumps cases in Poland, on lengthening of inter-epidemic period as well as on drift of infections towards older groups of children. The results of serological survey carried out on 1390 serum samples have indicated, that the proportion of positive serum samples (>11 VE/ml) was only 24.1% for children aged 1-4 years, 45.4% for children aged 5-9 years, 72.5% for age group 10-14 years, and over 85% for persons aged 15-30 years. Epidemiological data and the high proportion of individuals with negative titres of specific mumps IgG antibodies justify the need of introduction of obligatory mass immunisation against mumps in Poland. PMID- 15780718 TI - Peptide mimics as surrogate immunogens of mosquito midgut carbohydrate malaria transmission blocking targets. AB - Transmission blocking vaccines (TBV) against mosquito midgut carbohydrate epitopes is a promising approach to curbing the spread of malaria. However, carbohydrates as immunogens can be problematic. Via the malaria transmission blocking monoclonal antibody, MG96, we isolated dodecapeptide mimics of the conserved, nominal mosquito carbohydrate epitope from a peptide-display library. Two peptide clones, bearing a constrained, consensus motif competitively inhibited MG96 reactivity with its nominal midgut microvillar antigen. However, rabbit polyclonal antisera against these synthetic peptides recognized heterologous mosquito midgut carbohydrate and protein epitopes along the midgut basal lamina. Consequently, antisera did not block parasite development within the mosquito vector. Therefore, it is imperative that peptides not only need to be functional mimics but also complete mimotopes to effectively direct the vertebrate immune response towards the nominal, protective carbohydrate epitope on mosquito microvilli. PMID- 15780719 TI - Family practice nurse views on barriers to immunising children. AB - New Zealand (NZ) has low immunisation coverage for infants and children compared to many other westernised countries. Barriers to improving uptake are multifactorial, with health professional knowledge and attitudes identified as important modifiable factors. In NZ practice nurses give most childhood vaccinations in the primary health care setting. This study explored aspects of 150 family practice nurse views, knowledge and experience about immunisation. Qualitative and quantitative text data were obtained through randomised computer assisted telephone surveys and converged using a triangulated multi-method approach. Response rate was 89.3% nurses identified parents' fear as the greatest barrier to achieving better immunisation uptake and disagreed that health professional knowledge was a barrier. However, findings showed lack of knowledge among many participants, despite many feeling confident about their knowledge base. Factors associated with lower practice coverage of infants under 2 years were poorer knowledge of contraindications to vaccination and lack of completion of vaccinator training, especially an update course. A high level of confidence, more years in practice, dedicated time to follow-ups were not associated with better coverage rates. Practice nurses may be unaware that their knowledge in some areas needs improving. A trained practice nurse appears to play a significant role in overcoming fears and maintaining high coverage rates in their practice. We conclude that strategies that focus on primary health care provider support and education are more likely to gain high coverage than those that are purely directed at overcoming access barriers. PMID- 15780720 TI - Homotopes affect primary and secondary antibody responses in poultry. AB - Activation of the innate immune system by different homotopes may direct the subsequent specific immune response, and as a consequence may have significant implications on vaccination. In this study, effects of i.v. administered lipopolysaccharide (LPS), and lipoteichoic acid (LTA), respectively, and s.c. administered Keyhole Limpet Hemocyanin (KLH) on total and isotype-specific (IgM and IgG) primary and secondary antibody responses of chickens to human serum albumin (HuSA) were determined. Similarly, effects of pre-treatment of the birds with the homotopes on primary antibody responses to rabbit gamma globulin (RGG) were measured. In addition, antigen-specific (HuSA and RGG) and mitogenic (concanavalin A, Con A) responses of peripheral blood leucocytes in vitro were also measured. Our findings confirm earlier observations that both LTA as well as LPS have immunomodulatory features in chickens, albeit in an opposite direction. LTA enhanced primary and to a minor degree secondary antigen-specific antibody titers, but LPS was found to suppress these responses. In RGG-primed birds, LPS enhanced a non-antigen-specific recall response to HuSA. In addition, KLH acted in an immunomodulatory role too, enhancing primary and secondary antibody responses, but suppressing non-specific cellular immunity in a non-antigen specific fashion. Possible mechanisms underlying our observations, and the role of LTA, LPS, and KLH in polarization of the specific antibody response in chickens are discussed. PMID- 15780721 TI - Protection of macaques against Mycobacterium tuberculosis infection by a subunit vaccine based on a fusion protein of antigen 85B and ESAT-6. AB - Various new tuberculosis (TB) vaccine candidates in combination with new delivery systems, including subunit vaccines, are currently being evaluated by a number of laboratories. One vaccine candidate that has shown promising protective capacity in mice and guinea pigs is a fusion of Ag85B and ESAT-6. In this study, we have investigated the efficacy of this Ag85B-ESAT-6 fusion protein vaccine in a non human primate model for TB. Vaccination of cynomolgus monkeys with the Ag85B-ESAT 6 fusion protein in two different adjuvant (DDA/MPL, AS02A) resulted in a reduction in bacterial number and/or lung pathology in animals challenged with Mycobacterium tuberculosis. Vaccination prevented an increase in C-reactive protein serum levels, general activation of CD4 and CD8 subsets and boosted development of humoral and cellular immune responses to a spectrum of mycobacterial antigens on exposure to M. tuberculosis infection. We show, in two independent experiments, that vaccination of primates with Ag85B-ESAT-6 induces protective immune responses, suggesting that Ag85B-ESAT-6 is a strong candidate for further clinical evaluation. As far as we are aware this is the first report of protection in primates with a subunit vaccine. PMID- 15780722 TI - Vaccination against fowl cholera with acapsular Pasteurella multocida A:1. AB - We have previously constructed an acapsular Pasteurella multocida X-73 (serogroup A) mutant strain which was attenuated in virulence for chickens (Chung JY, Wilkie IW, Boyce JD, Townsend KM, Frost AJ, Ghodussi M, Adler B. Role of capsule in the pathogenesis of fowl cholera caused by Pasteurella multocida serogroup A. Infect. Immun. 2001;69:2487-2492). In this study, we have assessed the ability of this acapsular strain (PBA930) to induce protection against wild-type challenge in mice and the natural host chickens. Intramuscular administration of PBA930 to mice stimulated significant protection against X-73 and the heterologous strain P 1059 (A:3), but not against challenge with P-1662 (A:4). No protection was observed when PBA930 was introduced by the intraperitoneal or subcutaneous routes in mice. Significantly, the acapsular strain PBA930 was able to induce protection against challenge with wild type X-73 in chickens. PMID- 15780723 TI - Assessment of vaccine coverage following the introduction of a publicly funded pneumococcal vaccine program for the elderly in Victoria, Australia. AB - In 1998, a publicly funded pneumococcal vaccine program was introduced for persons aged > or = 65 years in Victoria, Australia. In 2000, vaccine coverage over the previous 5 years was assessed through a telephone survey of 385 randomly selected subjects aged > or = 65 years. Self-reported pneumococcal vaccine coverage within the previous 5 years was 46.0% (95% CI 40.5-51.6). Self-report was validated against the medical records of the nominated provider for 278 (72%) subjects. Among this subgroup, self-reported coverage was 51.1% but was 57.9% according to medical records (positive predictive value 91.5%, negative predictive value 77.2%, sensitivity 80.7%, specificity 89.7%). After accounting for response bias among those subjects for whom self-report was not validated, the revised estimate of pneumococcal vaccine coverage within the previous 5 years was 50.5% (95% CI 44.8-56.1). Comparison of vaccine coverage over time suggests the introduction of the publicly funded program in Victoria has dramatically increased coverage among the elderly. PMID- 15780724 TI - Policymakers' views regarding the introduction of new-generation vaccines against typhoid fever, shigellosis and cholera in Asia. AB - Face-to-face interviews and meetings with more than 160 policymakers and other influential professionals in seven large Asian countries (Bangladesh, China, India, Indonesia, Pakistan, Thailand and Vietnam) were conducted to survey opinions regarding the need for, and potential uses of new-generation vaccines against cholera, typhoid fever and shigellosis. Despite several barriers to their uptake--notably uncertainty of the burden of enteric diseases; preference for water, sanitation and other environmental improvements over vaccination for disease control; and high prices of the current vaccines relative to basic EPI vaccines, and their moderate protection levels--considerable interest was found in the targeted use of Vi typhoid vaccine in most countries, followed by (future) Shigella and oral cholera vaccines. The introduction of these vaccines in Asia could be greatly facilitated by country-specific evidence of disease burden, local or regional vaccine production, field studies demonstrating their safety and efficacy in local populations, evidence of potential economic savings from vaccination, and effective dissemination of research results to all those who make or influence immunization policy. PMID- 15780725 TI - Mucosal vaccination against diphtheria using starch microparticles as adjuvant for cross-reacting material (CRM197) of diphtheria toxin. AB - Mucosal vaccination has the advantage of eliciting a local mucosal immune response as well as a systemic response. In this investigation, polyacryl starch microparticles were conjugated to diphtheria toxin cross-reacting material (CRM197) as a mucosal adjuvant for oral or intranasal immunisation of mice. Various methods of stabilising CRM197 with formaldehyde were investigated. A good systemic and local mucosal immune response was attained with oral immunisation when CRM197 was treated with a relatively low formaldehyde concentration prior to conjugation to the microparticles. No immune response was seen after intranasal immunisation. PMID- 15780726 TI - Brucella spp. lumazine synthase: a novel antigen delivery system. AB - Lumazine synthase from Brucella spp. (BLS) was evaluated as a protein carrier to improve antigen delivery of KETc1, one of the peptides of the anti-cysticercosis vaccine. KETc1 becomes antigenic, preserved its immunogenicity and its protective capacity when expressed as a recombinant chimeric protein using Brucella spp. lumazine synthase. KETc1 and BLS-KETc1 were not MHC H-2(d), H-2(k) nor H-2(b) haplotype-restricted albeit KETc1 is preferentially presented in the H-2(b) haplotype. These findings support that BLS is a potent new delivery system for the improvement of subunit vaccines. PMID- 15780727 TI - Novel baculovirus-derived p67 subunit vaccines efficacious against East Coast fever in cattle. AB - Two novel baculovirus-derived recombinant Theileria parva p67 constructs were tested for their vaccine potential against East Coast fever. Boran calves were immunized with a his-GFP-p67 fusion protein (GFP:p67deltaSS) or with GP64:p67C, a protein fusion between a C-terminal domain of p67 and the baculovirus envelope protein GP64. Both GFP:p67deltaSS and GP64:p67C induced antibodies with high ELISA titers that neutralized T. parva sporozoites with high efficiency. Upon challenge, a correlation was observed between the in vitro neutralizing capacity and the reduction in severe ECF for individual animals. A protection level upto 85% was obtained. This level of protection was achieved with only two inoculations of 100 microg per dose, which is a major improvement over previous recombinant p67 products. PMID- 15780728 TI - A strong CD8+ T cell response is elicited using the synthetic polypeptide from the C-terminus of the circumsporozoite protein of Plasmodium berghei together with the adjuvant QS-21: quantitative and phenotypic comparison with the vaccine model of irradiated sporozoites. AB - Stable protective immunity can be achieved against malaria by the injection of radiation-attenuated sporozoites (gamma-spz) and is mediated by IFN-gamma producing CD8+ T cells targeting the pre-erythrocytic stages. An efficient malaria vaccine should mimic this immunity. We compared the immune response specific for the circumsporozoite protein (CSP) of Plasmodium berghei (P. berghei), an important target of this protective response, elicited in mice immunized with the long synthetic polypeptide (LSP) PbCS 242-310, representing the C-terminus of the CSP of P. berghei, with the adjuvant QS-21 or injected with gamma-spz. The ex vivo evaluation of the CD8+ T cell response by IFN-gamma ELISPOT assay revealed that the injection of LSP with QS-21 induced, compared to gamma-spz, a similar frequency of peptide-specific lymphocytes in the spleen but a eight-fold increase in the draining lymph-nodes. A very high frequency of CD8+ T cells, specific for the sequence PbCS 245-253, a H-2Kd-restricted CTL epitope, was obtained in the liver and spleen of mice immunized with the two regimens. Even though the frequency of H-2Kd PbCS 245-253 multimer+, CD8+ T cells was higher in gamma-spz immunized mice, the frequency of IFN-gamma producing CD8+ T cells was comparable. The phenotype of the CD8+ T cell responses was characterized with the help H-2Kd PbCS 245-253 multimer and most of the CSP specific CD8+ T cells represented an intermediate subset between effector and central memory with CD44(high), CD45RB(high), CD62L(low) and CD122(high). The number of memory CD8+ T cells decreased after the last LSP immunization but could be boosted to higher level with live spz. The unique combination of LSP PbCS 242 310 and the adjuvant QS-21 induced an immune response that was comparable in terms of quality to the one generated with gamma-spz. This confirmed the potential of LSP as malaria vaccine candidates as well as for the study of the repertoire of targets of protective immunity in the gamma-spz vaccine model. PMID- 15780729 TI - Establishment of Schistosoma japonicum calpain-specific mouse T cell hybridomas and identification of a T cell epitope that stimulates IFNgamma production. AB - Calpain is a calcium-dependent cystein protease, and the homologues of schistosome are known as one of vaccine candidate molecules against schistosomiasis. Here, we established two IL-2 producing T cell hybridoma cell lines specific for Schistosoma japonicum calpain, to identify T cell epitope(s) on the molecule. Overlapping 15mer oligopeptides of calpain were synthesized and tested for their stimulatory abilities to the hybridomas. As a result, epitopes recognized by the two hybridoma lines were the same: EQLKIYAQRC. Spleen cells from calpain multiple antigenic peptide (MAP)-immunized BALB/c mice produced IFNgamma upon stimulation with MAP or soluble worm antigen preparation (SWAP). The identification of the T cell epitope to stimulate Th1 response will contribute to the proper design of synthetic vaccines, evaluation of their protective potentials and elucidation of protective mechanisms in murine experimental schistosomiasis. PMID- 15780730 TI - Immunisation status in the public care system: a comparative study. AB - Children in public care have poor health outcomes despite statutory health assessments. Incomplete immunisation of children entering the care system has been reported. Does this health disadvantage persist for those established in the care system? The immunisation status of 119 children in public care for at least 6 months was compared to that noted in 119 age and sex matched children living in their own homes. Children in public care were significantly less likely to have received immunisations against diphtheria, tetanus, pertussis and polio, than the comparison group. This represents a persisting health disadvantage, which requires remedial action. PMID- 15780731 TI - Protection against experimental visceral leishmaniasis infection in dogs immunized with purified excreted secreted antigens of Leishmania infantum promastigotes. AB - The capacity of naturally excreted secreted antigens easily purified from culture supernatant of Leishmania infantum promastigotes (LiESAp), successfully cultivated in completely defined medium called CDM/LP [Lemesre JL. Methods for the culture in vitro of different stages of tissue parasites. International publication WO 94/26899, 1994; Merlen T, Sereno D, Brajon N, Rostand F, Lemesre JL. Leishmania spp: completely defined medium without serum and macromolecules (CDM/LP) for the continuous in vitro cultivation of infective promastigote forms. Am J Trop Med Hyg 1999;60(1):41-50] to protect dogs against experimental L. infantum infections is described. Eighteen healthy Beagle dogs were allocated into four groups that received at a 3-week interval either two subcutaneous injections of 50 microg (group 2, n = 3), 100 microg (group3, n = 6) and 200 microg (group 4, n = 3) LiESAp in formulation with muramyl dipeptide (MDP) or similar injections of placebo (group 1, n = 6). Dogs were intravenously infected with 10(8) metacyclic L. infantum promastigotes. Promastigotes of the MHOM/MA/67/ITMAP-263 and MHOM/FR/78/LEM75 strains were, respectively, administered 2 months (at day 84, homologous challenge 1) and 8 months post immunization (at day 273, heterologous challenge 2). The data indicated that vaccine candidate confers total protection (100%) against challenges 1 and 2 in dogs from groups 3 and 4 and intermediate protection (66.7%) against challenge 1 in dogs from group 2 as determined by parasite detection in bone marrow aspirates during 14 months post-challenge follow-up. All placebo dogs of group 1 were found infected and failed to respond to LiESAp in cell-mediated assays before and after both challenges. Increased levels of total anti-leishmanial antibodies were exclusively detected in infected dogs from group 1. Vaccine-induced protection correlates with an early establishment of a long lasting predominantly Th1-type cellular immune response specifically directed against LiESAp before and after experimental infections, as demonstrated by: (i) anti-LiESAp IgG2 reactivity, and (ii) LiESAp-specific lymphocyte proliferation assays and enhanced NO-mediated anti-leishmanial activity of canine monocyte-derived macrophages (CM-DM) in response to higher IFNgamma production by T-cells, when L. infantum-infected CM DM were co-cultured with autologous lymphocytes. Overall, our results support the view that a LiESAp vaccine might be useful in a promising vaccination approach against natural L. infantum infection. PMID- 15780732 TI - Adverse events following hexavalent vaccine (Hexavac): knowledge and informative sources. AB - This report describes adverse events to vaccination in eight children in southern Italy. These children were seen in the Emergency Room of a hospital of the Local Health Unit SA2 (Salerno, Italy) after administration of hexavalent vaccine (Hexavac, Aventis-Pasteur). On analysing available documentation on the adverse reactions (label instructions, Medline), the Authors concluded the cases reported were to be expected. However, the label instructions did not give sufficient information on the type and frequency of adverse reactions. If the label instructions are to furnish complete, operative information for the vaccination units, the Italian instructions for the Hexavac vaccine can be considered inadequate. PMID- 15780733 TI - Influenza vaccine in healthy children: a meta-analysis. AB - We conducted a meta-analysis of 13 randomised clinical trials evaluating the efficacy of influenza vaccine in healthy children. Against culture-confirmed influenza the overall efficacy was 74% (95% confidence interval, CI, 57%-84%), 65% for inactivated and 80% for live-attenuated vaccine. Corresponding figures were 59% (95% CI 43%-71%), 63% and 54% for serologically-confirmed influenza, and 33% (95% CI 29%-36%), 33% and 34% for clinical illness. Influenza vaccine is effective in preventing laboratory-confirmed and clinical influenza in healthy children, with no clear difference between inactivated and live-attenuated vaccine. Data on infants and younger children were too scanty to allow separate assessment. PMID- 15780734 TI - Decrease in pathology and progression of scrapie after immunisation with synthetic prion protein peptides in hamsters. AB - Effective therapy for prion diseases is currently unavailable. Recently, vaccination was shown to be effective in mouse models of a particular neurodegenerative conditions: Alzheimer's disease (AD). Here, we report that vaccination with synthetic oligopeptides homologous to the hamster (Mesocricetus auratus) prion protein augments survival time in animals infected intraperitoneally with 263K scrapie agent. For each hamster included in the study, prion-specific serum antibodies as well as deposition of pathological prion protein (PrP(res)), glial fibrillary acidic protein (GFAP), and mRNA expression for cytokines (TNF alpha, IL-1beta, IL-10) in brain tissues were evaluated. In immunized animals, increased survival after challenge was associated with a reduction of cerebral lesion, PrP deposition and GFAP expression; in these animals, anti-prion protein peptide antibody levels were increased, and the expression of pro-inflammatory cytokines (TNF alpha and IL 1beta) was reduced. Vaccination could be an effective therapeutic approach to postpone disease onset. PMID- 15780735 TI - Fusion of DsbA to the N-terminus of CTL chimeric epitope, F/M2:81-95, of respiratory syncytial virus prolongs protein- and virus-specific CTL responses in Balb/c mice. AB - In an effort to seek a means of inducing long lasting respiratory syncytial virus specific CTL responses in mice, we constructed a new recombinant protein, DsbA F/M2:81-95, by fusing carrier protein DsbA (disulfide bond isomerase) to the N terminus of CTL chimeric epitope F/M2:81-95 of this virus. DsbA-F/M2:81-95 can induce effectively virus-specific CTL responses as well as protective immunity without association with enhanced disease. Furthermore, compared with F/M2:81-95 alone, it increases the longevity of CTL responses in vivo up to 2.93 folds. Our study emphasizes that appropriate stimulation of non-antigen-specific T helper cells is essential to induce long lasting CD8+ CTL, and also implies DsbA-F/M2:81 95 may be a promising candidate for RSV vaccine development since it is an efficacious and safe immunogen. PMID- 15780736 TI - A two-dose schedule for combined hepatitis A and B vaccination in children aged 6 15 years. AB - A combined hepatitis A and B vaccine, Twinrix, in a paediatric formulation for ages 1-15 years and in an adult formulation for those ages 16 years and older, became commercially available in Turkey as well as in many countries. It is administered according to a three-dose schedule (0, 1 and 6 months). A reduction in the number of doses would improve the compliance rate and reduce administration costs. Therefore, we planned a trial evaluation of the immunogenicity, safety and reactogenicity profile of a high-dose combined hepatitis A and B vaccine, administered in two doses, compared with the profile of a paediatric-dose combined vaccine, administered in three doses, in healthy children aged 6-15 years. One hundred children were randomly attributed to the two study groups. The first group (paediatric-dose vaccine group) received the licensed Twinrix Paediatric, at months 0, 1 and 6; the second group (high-dose vaccine group) received the high-dose vaccine, following a 0, 6 months schedule. The reactogenicity was assessed after each vaccine dose. The immunogenicity was evaluated by testing for anti-HBs and anti-HAV antibodies. Seroconversion rates and geometric mean titres (GMTs) were compared. Both formulations of the combined vaccine were well tolerated. The high-dose combined vaccine administered in two doses, elicits satisfactory immunogenicity profiles, similar to those elicited by the paediatric vaccine administered in three doses. On completion of the vaccination schedule in the two groups all children were protected against hepatitis B and immune for hepatitis A. Anti-HAV GMTs after completion of the vaccination schedule were 7163 mlU/ml in the paediatric-dose group, 8241 mlU/ml in the high-dose group; anti-HBs GMTs were 8679 and 4583 mlU/ml, respectively. These results indicate that a two-dose schedule, compared with the standard three dose schedule, offers fewer injections for satisfactory protection against the two infections. This means fewer clinic visits, lower administration costs, better compliance, and higher coverage rate. Therefore, this two-dose schedule can be considered an appropriate regimen for the immunization of children and adolescents against hepatitis A and B infection, in the context of school-based immunization programmes. PMID- 15780737 TI - Analysis of hepatitis E virus neutralization sites using monoclonal antibodies directed against a virus capsid protein. AB - The dimeric form of the recombinant peptide (E2), comprising amino acid 394-606 of the capsid protein of hepatitis E virus (HEV), is strongly recognized by HEV reactive human serum, and when used as a vaccine, it protects rhesus monkeys against experimental HEV infection. In this work, the relationship of E2 to HEV has been probed using three murine monoclonal antibodies, 8C11, 13D8 and 8H3, all of which react predominantly against the E2 dimer, and can effect immune capture of the virus as well. 8C11 and 8H3 were further found to neutralize HEV infectivity in animals. Cross-blocking patterns between these antibodies discerned two spatially separate antigenic domains, one identified by 8C11 and 13D8, and the other, by 8H3. Kinetic studies using BIAcore biosensor suggest that the epitope to which 8H3 is directed is partially masked, and thus has limited access by the native antibody. However, this is not the case with the smaller Fab. Access to the 8H3 epitope was enhanced by the binding of 8C11, and inhibited by the binding of 13D8 to a distal site on the peptide. Similar to the effects of binding 8H3 to E2, 8C11 was found to enhance immune capture by 8H3, while 13D8 was inhibitory. Moreover, 8C11 and 8H3 act synergistically to neutralize HEV infectivity. The parallel cross-reaction patterns that these antibodies exhibit against the peptide and the virus, respectively, implicate two interacting conformationally dependent neutralization sites on the HEV particle. These sites might cooperate in the adsorption and penetration of the HEV virus. PMID- 15780738 TI - A bacterially expressed particulate hepatitis E vaccine: antigenicity, immunogenicity and protectivity on primates. AB - It was evaluated its antigenicity, immunogenicity and efficacy of a candidate recombinant hepatitis E virus (HEV) vaccine, referred hitherto as HEV 239 vaccine. The vaccine peptide has a 26 amino acids extension from the N terminal of another peptide, E2, of the HEV capsid protein, which has been shown to protect monkeys against HEV infection previously. The vaccine peptide is similar as E2 in that: first, the vaccine peptide migrates predominantly as dimer in SDS PAGE and it is dissociated into monomers by heating; second, its dimeric form of which predominantly recognized by HEV reactive human serum; and third, it shows the same pattern of reaction as E2 with a panel of eight monoclonal antibodies that had been raised against E2. In contrast to E2, the vaccine peptide aggregates to form particles of 13 nm mean radius, and consequently, it is more than 240 times more immunogenic than E2. Using alum as adjuvant, immunizing dose determined in mice was 80-250 ng for the vaccine and >60 microg for E2. Rhesus monkeys twice vaccinated with a 10 microg or a 20 microg formulation of this vaccine showed essentially the same antibody response, whereas the response to a 5 microg formulation was delayed but reached similar antibody levels. All the three vaccine formulations afford complete protection against infection with 10(4) genome equivalent dose of the homologous genotype 1 virus. At higher virus dose of 10(7), the same vaccine formulation partially protected against the infection and completely protected against hepatitis. The efficacy of the vaccine was essentially the same for the homologous genotype 1 virus and heterologous genotype 4 virus. PMID- 15780739 TI - Randomized trial of recombinant hepatitis B vaccine in HIV-infected adult patients comparing a standard dose to a double dose. AB - Because HIV and hepatitis B virus share many common risk factors, it is important to try to vaccinate HIV patients against hepatitis B. There are numerous reports describing a variety of dose schedules, limited success and markers associated with impaired response to HBV vaccine in these individuals. All studies have been small in size making it difficult to draw conclusions within and between studies. The purpose of this study was to evaluate a double dose of hepatitis B vaccine under more definitive guidelines: double blinded, randomized, controlled, with numbers for statistical validity. Two hundred and ten HIV infected subjects received a standard dose (20 microg) or a double dose (40 microg) of recombinant hepatitis B vaccine IM 0, 1 and 6 months. Ninety-four receiving standard dose and 98 receiving double dose completed the study. The seroconversion rate (anti-HBs > or = 10 mIU/mL) was 47 and 34% for double dose and standard dose, respectively (p = 0.07). A statistically significant higher seroconversion rate was associated with double dose comparing with standard dose for patients with CD4 cell counts > or = 350 cells/mm3 (64.3% x 39.3%; p = 0.008) but made no difference to seroconversion in those with CD4 <350 (23.8% x 26.3%; p = 0.80). Double dose also improved seroconversion comparing with standard dose for patients with HIV viral load <10,000 copies/mL (58.3% x 37.3%; p = 0.01) but made no difference to seroconversion in those with HIV viral load > or = 10,000 copies/mL (16% x 17%; p = 0.7). Based on the results of this study, the best current strategy for hepatitis B vaccination in HIV patients would be to use a double dose as a primary series when the viral load is likely to be low and CD4> or = 350, when there is likely to be an adequate immune response. PMID- 15780740 TI - Comparative studies of Avipox-GM-CSF versus recombinant GM-CSF protein as immune adjuvants with different vaccine platforms. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a potent immune stimulant when administered with different vaccines. Optimal use of GM-CSF resides in its ability to act locally to stimulate the proliferation and maturation of professional antigen-presenting cells (APCs) (i.e., Langerhans' cells) at the injection site. GM-CSF was engineered into a replication incompetent recombinant avian (fowlpox) virus (rF-GM-CSF) and a single subcutaneous injection resulted in a sustained enrichment of activated dendritic cells within the regional draining lymph nodes. Those changes were attributed to local GM-CSF production at the injection site by rF-GM-CSF-infected cells. Studies were carried out in which mice were administered different types of beta galactosidase (beta-gal)-based vaccines--whole protein, peptide, recombinant poxviruses--and GM-CSF was administered either as a single injection of rF-GM-CSF or four daily bolus injections of the recombinant protein. The use of rF-GM-CSF either improved the immune adjuvant effect, as observed for poxvirus-based vaccines, or was equivalent to rGM-CSF, as observed with the beta-gal protein vaccine. It is important to note that with either the replication-competent (vaccinia) or replication-incompetent (fowlpox) vaccines expressing LacZ, strong CTL responses directed against beta-gal were induced only when rF-GM-CSF was used as the immune adjuvant. Engineering GM-CSF into a recombinant fowlpox virus offers an excellent vehicle for the delivery of this cytokine as an immune adjuvant with specific vaccine platforms. In particular, delivery of GM-CSF via the rF-GM-CSF construct would be preferred over bolus injections of rGM-CSF when used as an immune adjuvant with whole protein or recombinant poxvirus-based vaccines. The study underscores the importance of defining the appropriate delivery form of an immune adjuvant, such as GM-CSF, relative to the immunization strategy to maximize the host immune responses against a specific antigen. PMID- 15780741 TI - Protection afforded by intranasal immunization with the neuraminidase-lacking mutant of influenza A virus in a ferret model. AB - Protective efficacy of the intranasal immunization with the neuraminidase (NA) deficient mutant of the influenza A virus was investigated in ferrets. Despite the highly attenuated replication in vivo, the mutant completely protected the animals against the wild type virus challenge. When challenge was done with antigenic drift variants, significant reductions in the viral titers, inflammatory cell counts, and protein concentrations were observed in the nasal washes of the immunized animals. The genetically engineered NA-deficient mutant also protected animals against the challenge and induced humoral immune response against the foreign protein that replaced the NA. We conclude that the NA as antigen is dispensable in the live attenuated influenza virus vaccine and that the NA-lacking mutant can be used as a virus vector. PMID- 15780742 TI - DNA immunization against respiratory syncytial virus (RSV) in infant rhesus monkeys. AB - A DNA vaccine was tested in infant Rhesus macaques to evaluate its safety, immunogenicity and protective efficacy. Monkeys were vaccinated and challenged with a clinical isolate of human RSV. Vaccinated animals developed humoral and cellular responses following inoculation with plasmid DNA encoding the fusion (F) and nucleoprotein (N), from closely related bovine RSV. Vaccinated monkeys had decreased RSV in their lungs post-infection, and there was a qualitative difference in histopathology observed between vaccinated and unvaccinated animals. The combined result of safety and immunogenicity in a neonatal primate model is encouraging, suggesting the feasibility of DNA vaccines against RSV in infants. PMID- 15780743 TI - Generation of influenza vaccine viruses on Vero cells by reverse genetics: an H5N1 candidate vaccine strain produced under a quality system. AB - Human influenza vaccine reference strains are prepared as required when an antigenically new strain is recommended by WHO for inclusion in the vaccine. Currently, for influenza A, these strains are produced by a double infection of embryonated hens' eggs using the recommended strain and the laboratory strain PR8 which grows to high titre in eggs, in order to produce a high growth reassortant (HGR). HGRs are provided by WHO reference laboratories to the vaccine manufacturing industry which use them to prepare seed virus for vaccine production. The use of reverse genetics in preparing vaccine reference strains offers several advantages over the traditional method: (i) the reverse genetics approach is a direct rational approach compared with the potentially hit-or-miss traditional approach; (ii) reverse genetics will decontaminate a wild type virus that may have been derived in a non-validated system, e.g. a cell line not validated for vaccine purposes, or that may contain additional pathogens; (iii) at the plasmid stage, the HA can be engineered to remove pathogenic traits. The use of reverse genetics in deriving HGRs has been demonstrated by several laboratories, including its use in deriving a non-pathogenic reassortant strain from a highly pathogenic virus. In this report, we have advanced the use of reverse genetics by making use of a cell line acceptable for human vaccine production, by demonstrating directly the short time frame in which a reassortant virus can be derived, and by deriving a non-pathogenic pandemic vaccine reference virus in cells validated for vaccine production and under quality controlled conditions. PMID- 15780744 TI - A comprehensive radiation hybrid map of the bovine genome comprising 5593 loci. AB - A bovine whole genome 7000-rad radiation hybrid (RH) panel, SUNbRH(7000-rad), was constructed to build a high-resolution RH map. The Shirakawa-USDA linkage map served as a scaffold to construct a framework map of 3216 microsatellites on which 2377 ESTs were ordered. The resulting RH map provided essentially complete coverage across the genome, with 1 cR7000 corresponding to 114 kb, and a cattle human comparative map of 1716 bovine genes and sequences annotated in the human genome, which covered 79 and 72% of the bovine and human genomes, respectively. We then integrated the bovine RH and comparative maps with BAC fingerprint information in to construct a detailed, BAC-based physical map covering a reported 40-cM quantitative trait locus region for intramuscular fat or "marbling" on BTA 4. In summary, the new, high-resolution SUNbRH7000-rad, comparative, Shirakawa-USDA linkage, and BAC fingerprint maps provide a set of genomic tools for fine mapping regions of interest in cattle. PMID- 15780745 TI - Molecular cytogenetics discards polyploidy in mammals. AB - Polyploidy, the presence of more than two chromosome sets, is common in plants, but extremely rare in animals. The absence of polyploid organisms with well differentiated sex chromosomes suggests that the disruption of the dosage between autosomes and sex chromosomes is incompatible with normal development. Thus, the announcement in 1999 of tetraploidy in a mammal, the South American red vizcacha rat Tympanoctomys barrerae, provoked great interest, even though the definitive proof of tetraploidy, the presence of four copies of each chromosome, was never provided. Here we used classical and molecular cytogenetics to test the ploidy level of T. barrerae and demonstrate that only two copies of each chromosome are present in this karyotype. The red vizcacha rat is clearly diploid and the amplification and dispersion of repetitive sequences best explain the large genome size of this mammal. Thus, polyploidy in mammals remains as unlikely as it has always been. PMID- 15780746 TI - Intergenic transcripts in genes with phase I introns. AB - Nonsense-mediated mRNA decay (NMD) is a eukaryotic quality-control mechanism that detects and degrades aberrant transcripts prematurely terminating translation. NMD may be elicited by intergenic transcripts that contain premature termination codons (PTCs), but chimeric mRNAs of genes that have introns of identical phase would be predicted to lack PTCs and escape NMD. We examined intron phase I containing HLA class II genes for the presence of intergenic mRNAs and found an extraordinary diversity of correctly spliced and polyadenylated intergenic transcripts. They lacked a significant homology at the chimeric joins and had no PTCs. Their expression levels were very low and positively correlated with the expression of natural transcripts. In contrast, pair-wise mixtures of separately transcribed plasmids carrying full-length HLA-DQB1, -DQA1, -DRB1, and -DRA cDNAs produced only hybrid molecules that lacked canonical exon boundaries, had homologous chimeric joins, and occasionally contained PTCs, implicating in vitro artifacts generated by template switching of Taq polymerase and reverse transcriptase. A differential exon structure of hybrid molecules observed in vitro and in cellular RNA preparations suggests that intergenic mRNAs with canonical exon boundaries arise in vivo during exon joining and/or transcription. Since the observed intergenic mRNAs may encode mixed class II heterodimers that were previously shown to present antigens it will be interesting to determine functional properties of such molecules in future studies. PMID- 15780747 TI - Distinct expression profiles of Mecp2 transcripts with different lengths of 3'UTR in the brain and visceral organs during mouse development. AB - Four different transcripts of the Mecp2 gene can be distinguished by the length of the 3' untranslated region generated by usage of alternative polyadenylation sites. In situ hybridization analyses encompassing embryonic to 20-week postnatal age showed that transcripts are expressed in the central nervous system, with a progressive restriction during development culminating in localized strong expression in the cerebral cortex, olfactory bulb, hippocampal formation, and internal granule and Purkinje layer of the cerebellum. Real-time RT-PCR measurements of Mecp2 transcript levels showed variations with mouse age in two distinctive patterns that are unique to the central nervous system and the visceral organs, respectively. The 10-kb mRNA is the predominant form expressed in the brain in contrast to the shorter species expressed in the lung and liver. The developmental profile of Mecp2 mRNA highlights a potential tissue-specific function of the 3'UTR in the regulation of MeCP2 protein synthesis in response to the age-specific requirement of MeCP2 function during the life of the mouse. PMID- 15780748 TI - A humanized mouse model for a common beta0-thalassemia mutation. AB - Accurate animal models that recapitulate the phenotype and genotype of patients with beta-thalassemia would enable the development of a range of possible therapeutic approaches. Here we report the generation of a mouse model carrying the codons 41-42 (-TTCT) beta-thalassemia mutation in the intact human beta globin locus. This mutation accounts for approximately 40% of beta-thalassemia mutations in southern China and Thailand. We demonstrate a low level of production of gamma-globins from the mutant locus in day 18 embryos, as well as production of mutant human beta-globin mRNA. However, in contrast to transgenic mice carrying the normal human beta-globin locus, 4-bp deletion mice fail to show any phenotypic complementation of the knockout mutation of both murine beta globin genes. Our studies suggest that this is a valuable model for gene correction in hemopoietic stem cells and for studying the effects of HbF inducers in vivo in a "humanized" thalassemic environment. PMID- 15780749 TI - Identification and characterization of multiple isoforms of a murine and human tumor suppressor, patched, having distinct first exons. AB - Mutations in mouse and human patched (PTCH) genes are associated with birth defects and cancer. PTCH, a 12-pass transmembrane protein, is a receptor for Sonic hedgehog (Shh) signaling proteins. Shh proteins activate transcription of target genes, including PTCH, via GLI transcription factors. Here we identified seven and five isoforms of human and mouse PTCH mRNA, respectively, which are generated by the complex alternative use of five exons as the first exon (exons 1a to 1e in the 5'-to-3' order). Although expression profiles of these isoforms were highly variable among human tissues, three of them, PTCHa, PTCHb, and PTCHd, were predominantly expressed in most tissues, PTCHd being most ubiquitous. In contrast, PTCHb was always predominant and reached a maximum at E10.5 during mouse development. These three mRNA isoforms encode three PTCH proteins with distinct N-termini, PTCH(L), PTCH(M), and PTCH(S). The expression of these three isoforms was regulated by GLI transcription factors, and at least two functional GLI-binding sequences were identified, one in exon 1a and the other between exon 1a and exon 1b. PTCH(L) and PTCH(M) were equally active in terms of suppressing GLI-mediated transcription and inducing apoptosis. PTCH(S) protein (encoded by PTCHd), lacking the first transmembrane domain, was more unstable than the other two, resulting in a reduced activity. This study may shed light on the mechanism whereby a single PTCH gene plays a role in both tumor cell growth and embryonic development. PMID- 15780750 TI - Two novel genes, Gpr113, which encodes a family 2 G-protein-coupled receptor, and Trcg1, are selectively expressed in taste receptor cells. AB - To identify genes important for taste receptor cell function, we analyzed the sequences and expression patterns of clones isolated from a mouse taste receptor cell-enriched cDNA library. Here, we report the analyses of two novel genes, Gpr113 and Trcg1. Gpr113 encodes a G-protein-coupled receptor belonging to family 2B, members of which are characterized by having long N-terminal, extracellular domains. The predicted N-terminal extracellular domain of GPR113 contains 696 amino acids with two functional domains, a peptide hormone-binding domain and a G protein-coupled receptor proteolytic site. Expression analyses indicate that Gpr113 expression is highly restricted to a subset of taste receptor cells. TRCG1 is also selectively expressed in a subset of taste receptor cells. Trcg1 is alternatively spliced and encodes Trcg1 isoforms of 209 and 825 amino acids. BLAST searches of genomic sequences indicate that a putative homolog of Trcg1 resides on human chromosome 15q22. PMID- 15780751 TI - Characterization and expression analysis of the Spink5 gene, the mouse ortholog of the defective gene in Netherton syndrome. AB - The human SPINK5 gene, encoding the putative 15-domain serine protease inhibitor LEKTI, was identified as the defective gene in the severe autosomal recessive ichthyosiform skin disorder known as Netherton syndrome and as a candidate susceptibility gene for atopic disease. Here we report mapping of the murine Spink5 gene to chromosome 18 and its characterization. We show that, unlike in humans, transcription of the mouse Spink5 gene generates two mRNAs that differ in the 3' untranslated region. The encoded protein, which is detected in differentiated primary cultured keratinocytes and mouse skin as an approximately 130-kDa glycosylated precursor, displays approximately 60% identity with its human counterpart but lacks the human LEKTI domain 6. As in the human, mouse Lekti represents a marker of epithelial differentiation, strongly expressed in the granular layer of the epidermis, in suprabasal layers of stratified epithelia, and in thymic Hassall's bodies. Our data indicate that mouse Spink5/Lekti, like its human counterpart, is involved in the control of epithelial tissue homeostasis, but also highlight specific features of the murine gene and protein. PMID- 15780752 TI - Cloning of the murine ER71 gene (Etsrp71) and initial characterization of its promoter. AB - The ER71 protein belongs to the ETS transcription factor family and is testis specifically expressed in adult mice. Here we describe the cloning of the respective Etsrp71 gene and promoter. The murine Etsrp71 gene is relatively compact, spanning 3 kb, and is arranged into seven exons and six introns, the majority of which are highly conserved in rat and human. Its promoter is devoid of a TATA box and transcription starts at multiple sites. Furthermore, two ER71 isoforms exist that differ by 22 N-terminal amino acids, but show no difference in DNA binding or transactivation. Close to the transcription initiation sites, we identified a binding site for the transcription factor Sp1. Mutation of this binding site severely diminished the ability of Sp1 to activate the Etsrp71 promoter. The findings reported here may provide avenues for further research elucidating the regulation of Etsrp71 gene activity during embryogenesis and in adult testes. PMID- 15780753 TI - IRIS: a database surveying known human immune system genes. AB - We have compiled an online database of known human defense genes: the Immunogenetic Related Information Source (IRIS). As of October 1, 2004, there are 1562 immune genes recorded in IRIS, representing 7% of the human genome. This resource contains searchable information including chromosomal location, sequence data, and a curated functional annotation for each entry. We used IRIS as a basis for analyzing the composition and characteristics of the immune genome, such as gene clustering, polymorphism, and relationship to disease. High protein sequence similarity correlated inversely with distance between immune genes, consistent with clustering of duplicated loci. We also found that, even though some immune genes exhibit high levels of polymorphism, such as MHC class I, the range of levels of polymorphism in immune genes is similar to that of nonimmune genes. Approximately 20% of immune genes have a known disease association. IRIS is available online at . PMID- 15780754 TI - Automated characterization of potentially active retroid agents in the human genome. AB - Retroid agents are genomes that encode the reverse transcriptase (RT) and replicate by way of an RNA intermediate. Some retroid agents are implicated in disease via insertional mutagenesis, while others have been found to encode proteins essential to primate reproduction or provide regulatory sequences for host cell processes. The Genome Parsing Suite (GPS), a generic multistep automated process, was developed to characterize all RT-like sequences in the human genome database and to annotate the gene complement of the retroid agents that encode these sequences. In this report the GPS analyzes all significant WU tBLASTn hits returned for 30 representative RT queries. A total of 128,779 unique RT signals were identified, and 7594 of these were retrieved by RTs not previously reported in the human genome. We have identified 9652 full-length long interspersed nuclear elements (LINEs). Only 159 LINEs are without stop codons or frameshifts. PMID- 15780755 TI - The DUB/USP17 deubiquitinating enzymes, a multigene family within a tandemly repeated sequence. AB - Here we report the identification of 10 human, 1 murine, and 2 rat ORFs, all of which represent additional members of the DUB/USP17 family of deubiquitinating enzymes. In addition, we demonstrate that this family constitutes part of a tandemly repeated sequence conserved throughout humans, mice, and rats. Furthermore, upon examination of the known family members we have found that the multiple genes observed, in contrast to other gene families, have arisen due to the independent expansion of an ancestral sequence within each species. This premise is further strengthened by the observation that the murine and rat genes span two exons while their human counterparts have one. These observations, in conjunction with previous work demonstrating that the DUB/USP17's are cytokine inducible and that they regulate both cell growth and survival, suggest that the DUB/USP17's are a large highly conserved family of genes that may play an important role in controlling cell fate. PMID- 15780756 TI - HO in pregnancy. AB - The enzyme heme oxygenase (HO) has been implicated in several physiological functions throughout the body including control of vascular tone and regulation of the inflammatory and apoptotic cascades as well as contributing to the antioxidant capabilities in several organ systems. These various properties attributed to HO are carried out through the catalytic products of heme degradation, namely carbon monoxide (CO), biliverdin, and free iron (Fe2+). As the newly emerging roles of HO in normal organ function have come to light, researchers in several disciplines have assessed the role of this enzyme in various physiological and pathological changes taking place in the human body over a lifetime. Included in this new wave of interest is the involvement of HO, and its by-products, in the normal function of the vital organ of pregnancy, the placenta. In this review the role of HO, and its catalytic products, will be examined in the context of pregnancy. The different isoforms of the HO enzyme (HO 1, HO-2, HO-3) have been localized throughout placental tissue, and have been shown to be physiologically active. The HO protein and more specifically its catalytic by-products (CO, biliverdin, and Fe2+) have been postulated to be involved in the maintenance of uterine quiescence throughout gestation, regulation of hemodynamic control within the uterus and placenta, regulation of the apoptotic and inflammatory cascades in trophoblast cells, and the maintenance of a balance of the oxidant-antioxidant status within the placental tissues. The association between this enzyme system, and its above-noted roles throughout pregnancy, with the hypertensive disorder of pregnancy preeclampsia (PET), will also be examined. It is hypothesized that a decrease in HO expression and/or activity throughout gestation would be capable of initiating several pathological processes involved in the etiology of PET. This hypothesis has led to further discussion emphasizing the possibility of novel therapeutic designs targeting this enzyme system for the treatment of PET. PMID- 15780757 TI - Nox 2 stimulates muscle differentiation via NF-kappaB/iNOS pathway. AB - The NF-kappaB/iNOS pathway stimulates muscle differentiation downstream of the PI 3-kinase/p38 MAPK pathway and diverse antioxidants block muscle differentiation. Therefore, we here investigated whether Nox 2 links those two myogenic pathways in H9c2 and C2C12 myoblasts. Compared with the proliferation stage, ROS generation was enhanced from the early stage of differentiation and gradually increased as differentiation progressed. Antioxidants suppressed the activated NF kappaB/iNOS pathway during muscle differentiation. Nox 2 activity was also increased during muscle differentiation. Treatment with DPI and apocynin, two inhibitors of NADPH oxidase, and suppression of Nox 2 expression using siRNA, but not Nox 1, inhibited NADPH oxidase activity, muscle differentiation, and the NF kappaB/iNOS pathway. Inhibition of PI 3-kinase and p38 MAPK suppressed the Nox 2/NF-kappaB/iNOS pathway. Nitric oxide restored muscle differentiation blocked by treatment with antioxidants or suppression of the Nox 2/NF-kappaB/iNOS pathway. In conclusion, Nox 2 stimulates muscle differentiation downstream of the PI 3 kinase/p38 MAPK pathway by activating the NF-kappaB/iNOS pathway via ROS generation. PMID- 15780759 TI - Chemiluminescence of Pholasin caused by peroxynitrite. AB - The kinetics of the oxidation of Pholasin by peroxynitrite, which leads to emission of light, were studied. The reaction shows a lag phase, which is smaller at higher peroxynitrite-to-Pholasin ratios. The total light emission approximately doubles from pH 5 to 9 and decreases precipitously to half the pH 5 value at pH 10. Dioxygen and carbon dioxide accelerate the reaction course, but they do not change significantly the reaction yield. Chemiluminescence of Pholasin is suppressed by antioxidants, but no significant shift is noticed in the time at which light emission is maximal. The chemiluminescence intensity is strongly dependent on the potassium concentration, although it is not significantly affected by lithium, cesium, or magnesium; potassium decreases luminescence. The mechanism of the peroxynitrite-induced oxidation of Pholasin may start with the reversible formation of a protein-peroxynitrite intermediate, or a first oxidation product, followed in subsequent steps by decomposition and light emission. However, many questions concerning the mechanism of the light emission remain to be elucidated. PMID- 15780758 TI - Stimulation of HIF-1alpha, HIF-2alpha, and VEGF by prolyl 4-hydroxylase inhibition in human lung endothelial and epithelial cells. AB - Diminished alveolar and vascular development is characteristic of bronchopulmonary dysplasia (BPD) affecting many preterm newborns. Hypoxia promotes angiogenic responses in developing lung via, for example, vascular endothelial growth factor (VEGF). To determine if prolyl 4-hydroxylase (PHD) inhibition could augment hypoxia-inducible factors (HIFs) and expression of angiogenic proteins essential for lung development, HIF-1alpha and -2alpha proteins were assessed in human developing and adult lung microvascular endothelial cells and alveolar epithelial-like cells treated with either the HIF PHD-selective inhibitor PHI-1 or the nonselective PHD inhibitors dimethyloxaloylglycine (DMOG) and deferoxamine (DFO). PHI-1 stimulated HIF-1alpha and -2alpha equally or more effectively than did DMOG or DFO, enhanced VEGF release, and elevated glucose consumption, whereas it was considerably less cytotoxic than DMOG or DFO. Moreover, VEGF receptor Flt-1 levels increased, whereas KDR/Flk-1 decreased. PHI-1 treatment also increased PHD-2, but not PHD-1 or -3, protein. These results provide proof of principle that HIF stimulation and modulation of HIF-regulated angiogenic proteins through PHI-1 treatment are feasible, effective, and nontoxic in human lung cells, suggesting the use of PHI 1 to enhance angiogenesis and lung growth in evolving BPD. PMID- 15780760 TI - In vivo protection of synaptosomes from oxidative stress mediated by Fe2+/H2O2 or 2,2-azobis-(2-amidinopropane) dihydrochloride by the glutathione mimetic tricyclodecan-9-yl-xanthogenate. AB - D609 (tricyclodecan-9-yl-xanthogenate) is a phosphatidylcholine-specific phospholipase C inhibitor that also has been reported to protect rodents against oxidative damage caused by lethal doses of ionizing radiation. We previously showed that D609 mimics glutathione. D609 has a free thiol group, which upon oxidation forms a disulfide. The resulting dixanthate is a substrate for glutathione reductase, regenerating D609. Recent studies from our laboratory have also shown that D609 reduces the Alzheimer amyloid beta-peptide (1-42)-induced oxidative stress and cytotoxicity in neuronal cell culture. The present study was undertaken to test the hypothesis that D609 would provide neuroprotection against free radical oxidative stress in vivo. Synaptosomes isolated from gerbils, previously injected intraperitoneally (ip) with D609, were treated with the oxidants Fe2+/H2O2 or 2,2-azobis-(2-amidinopropane) dihydrochloride (AAPH), which produce free radicals. Synaptosomes isolated from the gerbils ip injected with D609 and treated with Fe2+/H2O2 or AAPH showed significant reduction in reactive oxygen species, levels of protein carbonyl, protein-bound hydroxynonenal (a lipid peroxidation product), and 3-nitrotyrosine (another marker of protein oxidation formed by reaction of tyrosine residues with peroxynitrite) compared to oxidative stress in synaptosomes isolated from gerbils that were injected with saline, but treated with Fe2+/H2O2 or AAPH. These results are discussed with reference to the potential use of this brain-accessible glutathione mimetic in the treatment of oxidative stress-related neurodegenerative disorders. PMID- 15780761 TI - Urinary 20-hydroxyeicosatetraenoic acid excretion is associated with oxidative stress in hypertensive subjects. AB - Oxidative stress has been implicated in the pathogenesis of hypertension. 20 Hydroxyeicosatetraenoic acid (20-HETE) is a cytochrome P450 product of arachidonic acid metabolism, thought to be involved in the regulation of blood pressure (BP). The metabolism of arachidonic acid by cytochrome P450 enzymes may be a significant source of oxidative stress. F2-isoprostanes are reliable markers of in vivo oxidative damage. gamma-Glutamyl transpeptidase (gamma-GT) has traditionally been associated with alcohol intake or liver dysfunction and may be an early marker of oxidative stress. The objective of the present study was to investigate relationships between 20-HETE excretion and markers of oxidative stress (F2-isoprostanes and gamma-GT). Sixty-nine treated hypertensive subjects underwent measurement of 24-h ambulatory BP, serum gamma-GT, and urinary F2 isoprostane and 20-HETE excretion. 20-HETE excretion was positively associated with 24-h diastolic BP (p = 0.005), alcohol intake (p = 0.008), gamma-GT (p = 0.007), and F2-isoprostanes (p = 0.005). F2-isoprostanes were positively associated with alcohol intake (p = 0.018) and gamma-GT (p = 0.01). In a multivariate regression, gamma-GT remained an independent predictor of 20-HETE excretion, after adjustment for age, gender, BMI, and alcohol intake. In conclusion, the study highlights the positive association observed between 20 HETE excretion and markers of oxidative damage. The study also provides evidence that gamma-GT may be a useful marker of oxidative stress. PMID- 15780762 TI - Spectroscopic properties and reactivity of free radical forms of A2E. AB - A pyridinium bisretinoid (A2E) is the only identified blue-absorbing chromophore of retinal lipofuscin that has been linked to its aerobic photoreactivity and phototoxicity. Pulse radiolysis has been used to study both the one-electron oxidation and the one-electron reduction of A2E in aqueous micellar solutions. The reduction to the semireduced A2E (lambda(max) broad and between 500 and 540 nm) was achieved with formate radicals and the subsequent decay of A2E* was slow (over hundreds of milliseconds) via complex kinetics. The long lifetime of the A2E* should facilitate its reactions with other biomolecules. For example, with oxygen, the A2E* produced the superoxide radical anion with a rate constant of 3 x 10(8) M(-1) s(-1). The A2E was also reduced by the NAD radical, the corresponding rate constant being 2.3 x 10(8) M(-1) s(-1). Other experiments showed that the one-electron reduction potential of A2E lies in the range -640 to -940 mV. The semioxidized form of A2E (lambda(max) 590 nm) was formed via oxidation with the Br2*- radical and had a much shorter lifetime than the semireduced form. With strongly oxidizing peroxyl radicals (CCl3O2*) our kinetic data suggest the formation of a radical adduct followed by dissociation to the semioxidized A2E. With milder oxidizing peroxyl radicals such as that from methanol, our results were inconclusive. In benzene we observed an efficient oxidation of zeaxanthin to its radical cation by the A2E radical cation; this may be relevant to a detrimental effect of A2E in vision. PMID- 15780764 TI - Regulation of apoptosis by glutathione redox state in PC12 cells exposed simultaneously to iron and ascorbic acid. AB - We previously reported that the levels of non-protein-bound iron (NPBI) and ascorbic acid (AA) are markedly increased in the cerebrospinal fluid of infants with perinatal asphyxia. The present study showed that FeSO4 and AA synergistically induced apoptosis of PC12 cells, which was prevented by alpha tocopherol and glutathione (GSH) ethyl ester. Markers of free radical damage, such as ortho-tyrosine, meta-tyrosine, and F(2alpha)-isoprostane, showed a gradual increase. AA and ferrous NPBI disappeared rapidly from the culture medium, but exposure for only a few hours was sufficient to trigger apoptosis. Intracellular GSH decreased progressively along with a concomitant increase of glutathione disulfide (GSSG). The baseline half-cell reduction potential (Ehc) for GSSG, 2H+/2GSH couple was -246 mV and an Ehc of -200 mV was the critical level to switch on apoptosis, although some cells escaped this fate by transient increase of intracellular GSH. Once Ehc reached around -165 mV (81 mV oxidation from the baseline), all cells lost the ability to maintain an adequate intracellular GSH level and subsequently underwent apoptosis. These findings at least partly explain the mechanism of Fe-AA cytotoxicity, in that ferrous iron catalyzes hydroxyl radical generation and induces lipid peroxidation, after which subsequent depletion of GSH raises Ehc to the critical level for triggering or potentiating the apoptotic cascade. PMID- 15780763 TI - CD36 overexpression in ritonavir-treated THP-1 cells is reversed by alpha tocopherol. AB - Therapies with antiretroviral protease inhibitors (ARPI) are correlated with a higher risk for dyslipidemia, hypercholesterolemia, and atherosclerosis. The original aim of this study was to establish whether alpha-tocopherol can reduce CD36 scavenger receptor overexpression occurring after treatment of monocytes with the ARPI ritonavir. We show here that treatment of THP-1 monocytes with ritonavir increases total protein and surface expression of CD36; however, only weak changes are observed at the mRNA level, suggesting that CD36 overexpression occurs mainly at the posttranscriptional level. Concentrations of ritonavir that upregulate CD36 expression inhibit proteasome activity in THP-1 cells, indicating a possible regulatory role of the proteasome in CD36 overexpression. Similar to ritonavir, the proteasome inhibitor ALLN increases the CD36 surface expression on THP-1 cells. alpha-Tocopherol efficiently normalizes CD36 protein overexpression after ritonavir treatment and reduces oxLDL uptake. Furthermore, in THP-1 monocytes, alpha-tocopherol reverses the proteasome activity inhibited by ritonavir. This study indicates that an increased CD36 protein expression in THP 1 monocytes induced by ritonavir can be normalized by alpha-tocopherol. CD36 overexpression is caused by inhibition of proteasome activity by ritonavir, which is efficiently restored by alpha-tocopherol. PMID- 15780765 TI - Arginase-1 overexpression induces cationic amino acid transporter-1 in psoriasis. AB - Regulated uptake of extracellular l-arginine by cationic amino acid transporters (CATs) is required for inducible nitric oxide synthase and arginase activity. Both enzymes were recently recognized as important in the pathophysiology of psoriasis because of their contribution to epidermal hyperproliferation. We here characterize the expression pattern of CATs in psoriatic skin compared to healthy skin. CAT-1 mRNA expression was strongly upregulated in lesional and nonlesional areas of psoriatic skin compared to healthy skin, whereas expression of CAT-2A and the inducible isoform CAT-2B was unaltered in psoriatic skin. Furthermore, we tested the hypothesis that arginase-1 overexpression regulates CAT expression via intracellular l-arginine concentration. In in vitro experiments with arginase-1 overexpressing HaCaT cells, CAT-1 mRNA expression was increased. Likewise, this occurs in l-arginine-starved HaCaT cells. Both CAT-2 isoforms were not affected. Arginase-1 overexpression limits the synthesis of NO at physiological, but not supraphysiological, l-arginine levels. Plasma l-arginine concentration was diminished in psoriasis patients and the arginase product l-ornithine was significantly increased compared to healthy controls. In summary, arginase-1 overexpression leads to upregulated CAT-1 expression in psoriatic skin, which is due to lowered intracellular l-arginine levels and limits NO synthesis at physiological l-arginine concentrations. PMID- 15780766 TI - Oxidative stress and lipid mediators induced in alveolar macrophages by ultrafine particles. AB - In ambient aerosols, ultrafine particles (UFP) and their agglomerates are considered to be major factors contributing to adverse health effects. Reactivity of agglomerated UFP of elemental carbon (EC), Printex 90, Printex G, and diesel exhaust particles (DEP) was evaluated by the capacity of particles to oxidize methionine in a cell-free in vitro system for determination of their innate oxidative potential and by alveolar macrophages (AMs) to determine production of arachidonic acid (AA), including formation of prostaglandin E2 (PGE2), leukotriene B4 (LTB4), reactive oxygen species (ROS), and oxidative stress marker 8-isoprostane. EC exhibiting high oxidative potential induced generation of AA, PGE2, LTB4, and 8-isoprostane in canine and human AMs. Printex 90, Printex G, and DEP, showing low oxidative capacity, still induced formation of AA and PGE2, but not that of LTB4 or 8-isoprostane. Aging of EC lowered oxidative potential while still inducing production of AA and PGE2 but not that of LTB4 and 8-isoprostane. Cellular ROS production was stimulated by all particles independent of oxidative potential. Particle-induced formation of AA metabolites and ROS was dependent on mitogen-activated protein kinase kinase 1 activation of cytosolic phospholipase A2 (cPLA2) as shown by inhibitor studies. In conclusion, cPLA2, PGE2, and ROS formation was activated by all particle types, whereas LTB4 production and 8 isoprostane were strongly dependent on particles' oxidative potential. Physical and chemical parameters of particle surface correlated with oxidative potential and stimulation of AM PGE2 and 8-isoprostane production. PMID- 15780767 TI - Aggregates of oxidized proteins (lipofuscin) induce apoptosis through proteasome inhibition and dysregulation of proapoptotic proteins. AB - Cellular senescence may be accompanied by accumulation of large aggregates of oxidized proteins, also known as lipofuscin. The hypothesis that cellular accumulation of lipofuscin-like materials (LIP) results in cell death as a result of proteasome inhibition was examined. Rat neonatal cardiomyocytes were incubated with synthetic LIP for up to 48 h. This was accompanied by increases in cellular autofluorescence (207% by 48 h; p < 0.05) and electron microscopic evidence of internalization of LIP particles. LIP incubation resulted in loss of viability ( 46% by 48 h; p < 0.05) through apoptotic cell death. Although 20S-proteasome activity was increased by 74% after 6 h, both 20S- and 26S-proteasome activities were decreased after 48 h of incubation (-54% (p < 0.05) and -50%, respectively), accompanied by large increases in ubiquitinated proteins. Several proteasome regulated proapoptotic proteins, including c-Jun (2.9-fold; p < 0.05), Bax (1.8 fold; p < 0.05), and p27(kip1) (3.2-fold; p < 0.05), were observed to be increased by 48 h. Observation of ubiquitinated homologues of Bax and p27(kip1) suggested that part of the increase was due to decreased proteasomal degradation of these proteins. The results of this study are consistent with the conclusion that accumulation of LIP results in inhibition of the proteasome, which initiates an apoptotic cascade as a result of dysregulation of several proapoptotic proteins. PMID- 15780768 TI - Nitric oxide-related species-induced protein oxidation: reversible, irreversible, and protective effects on enzyme function of papain. AB - Protein oxidation, irreversible modification, and inactivation may play key roles in various neurodegenerative disorders. Therefore, we studied the effects of the potentially in vivo occurring nitric oxide-related species on two different markers of protein oxidation: protein carbonyl generation on bovine serum albumine (BSA) and loss of activity of a cysteine-dependent protease, papain, in vitro by using Angeli's salt, papanonoate, SIN-1, and S-nitrosoglutathione (GSNO) as donors of nitroxyl, nitric oxide, peroxynitrite, and nitrosonium ions, respectively. Angeli's salt, SIN-1, and papanonoate (0-1000 microM) all generated a concentration-dependent increase in carbonyl formation on BSA (107, 60, and 45%, respectively). GSNO did not affect carbonyl formation. Papain was inhibited by Angeli's salt, SIN-1, papanonoate, and GSNO with IC50 values of 0.62, 2.3, 54, and 80 microM, respectively. Angeli's salt (3.16 microM)-induced papain inactivation was only partially reversible, while the effects of GSNO (316 microM) and papanonoate (316 microM) were reversible upon addition of excess DTT. The Angeli's salt-mediated DTT-irreversible inhibition of papain was prevented by GSNO or papanonoate pretreatment, hypothetically through mixed disulfide formation or S-nitrosylation of the catalytically critical thiol group of papain. These results, for the first time, compare the generation of carbonyls in proteins by Angeli's salt, papanonoate, and SIN-1. Furthermore, these results suggest that S-nitrosothiols may have a novel function in protecting critical thiols from irreversible oxidative damage. PMID- 15780769 TI - Overexpression of copper/zinc superoxide dismutase decreases ischemia-like astrocyte injury. AB - Overexpression of copper/zinc superoxide dismutase (SOD1) in transgenic mice protects from transient focal cerebral ischemia in adult animals, but increases oxidative injury in perinatal mice. The effect of SOD1 overexpression on astrocytes subjected to ischemia-like insults has not yet been determined. Overexpression of human SOD1 in astrocytes resulted in a 3-fold increase in SOD1 activity without coupled up-regulation of catalase or glutathione peroxidase activities. Cells subjected to oxygen-glucose deprivation (OGD) or glucose deprivation to mimic ischemic injury were protected by SOD1 overexpression. OGD injury was reduced 47.6+/-9.3%, assessed by release of lactate dehydrogenase. OGD also caused a significant increase in catalase activity which was moderated by SOD1 overexpression. The level of glutathione in astrocytes overexpressing SOD1 was maintained at higher levels following 5 h OGD compared to control cultures under the same conditions. Reduction of glutathione prior to OGD significantly increased cell death of SOD1-overexpressing astrocytes as well as controls, but SOD1 still provided significant protection, suggesting that both GSH-dependent scavenging and GSH-independent scavenging are relevant to SOD1 protection in astrocytes. PMID- 15780770 TI - Body mass index and cardiovascular disease in a birth cohort of commuter air carrier and air taxi pilots. AB - PURPOSE: Acute cardiovascular events such as heart attack and stroke in commercial pilots have been a concern to flight safety. The purpose of this study was to examine the association of body mass index (BMI) with cardiovascular disease in commercial pilots. METHODS: We followed from 1987 to 1997 a cohort of 3019 male commuter and air taxi pilots who were born between 1933 and 1942 and who were certified by the US Federal Aviation Administration. The relationship between BMI and the risk of cardiovascular disease was assessed using Poisson regression modeling based on generalized estimation equations. RESULTS: At baseline, 55% of the pilots were overweight (25 kg/m(2) BMI < 30 kg/m(2)) and 7% were obese (BMI >/= 30 kg/m(2)). The follow-up accumulated a total of 20,671 person-years and 1897 diagnoses of cardiovascular disease, yielding an incidence rate of 92 per 1000 person-years. The incidence rates of cardiovascular disease increased significantly with BMI. With adjustment for baseline history of cardiovascular disease and age, pilots who were overweight and obese had 6% and 22% higher risk of cardiovascular disease, respectively, than their counterparts with normal BMI. CONCLUSIONS: The prevalence of overweight/obesity and the relationship between BMI and cardiovascular disease among commercial pilots parallel findings from the general population. Incorporating BMI into the current medical standards for commercial pilots merits serious consideration. PMID- 15780771 TI - Accuracy of adult reported pneumococcal vaccination status of children. AB - PURPOSE: A pneumococcal conjugate vaccine was licensed in 2000 for use in children. To evaluate the accuracy of an adult's recall of children's vaccination status with the pneumococcal vaccine, the authors conducted a cross-sectional survey of adults in Philadelphia County from January to December 2003. METHODS: Subjects were identified by random digit dialing and interviewed regarding the age and vaccination status of all children 6 years old or younger with whom they had significant contact. The gold standard for determining the accuracy of vaccine status reports was the Kids Immunization Database/Tracking System (KIDS) at the City of Philadelphia Department of Public Health. RESULTS: One hundred twenty-two interviews were completed with vaccination data collected on 185 children. One hundred sixty-six children (90%) were matched with records in the KIDS registry. The sensitivity and specificity of adult reported child vaccine status were 0.65 (95% CI, 0.55, 0.74) and 0.47 (95% CI, 0.34, 0.61), respectively. Agreement between adult report and the KIDS registry was 59%. Significant variation in agreement was not found to be related to demographic characteristics of the adult subjects. CONCLUSION: Adult report of the pneumococcal vaccination status of children was found to be relatively inaccurate and did not vary by known demographic characteristics. PMID- 15780772 TI - Leisure time, non-leisure time, and occupational physical activity in Asian Americans. AB - PURPOSE: Asian American immigrants' risk of heart disease, diabetes, and obesity increase with duration of residence in the United States (US). Regular physical activity reduces the risk of these diseases, yet little is known about physical activity in Asian Americans and how it changes after immigration. METHODS: Data from the 2001 California Health Interview Survey, which oversampled Asian Americans, were analyzed to investigate the effects of ethnicity, nativity, and years in the US on leisure time physical activity (LTPA), non-leisure time physical activity (NLTPA), and occupational physical activity. A total of 4226 Asian Americans and 29,473 US-born non-Asians were included. RESULTS: Asian Americans were much less likely to meet recommended levels of LTPA than US-born non-Asians (odds ratio [OR], men=0.51, 95% confidence interval [CI], 0.42, 0.61, OR, women = 0.48, 95% CI, 0.40, 0.57). Foreign-born Asians were least likely to participate in LTPA; LTPA increased as years in the US increased. After accounting for NLTPA, Asian Americans had significantly lower estimated weekly energy expenditure than US-born non-Asians. CONCLUSIONS: Asian Americans, especially immigrants, are at risk for low levels of LTPA and high levels of physical inactivity. NLTPA does not offset these lower levels of LTPA. Increasing physical activity is key to protecting the health of this rapidly growing population. PMID- 15780773 TI - Counts of neutrophils, lymphocytes, and monocytes, cause-specific mortality and coronary heart disease: the NHANES-I epidemiologic follow-up study. AB - PURPOSE: To examine the association of elevated counts of white blood cell types with increased risk of coronary heart disease (CHD) and death. METHODS: Data were examined from the NHANES-I Epidemiologic Follow-up Study. RESULTS: Relative risks for death at ages 25 to 74 comparing the upper and lower tertiles of neutrophil count were: all causes 1.29 (95% CL, 1.14, 1.47), and cardiovascular causes 1.39 (95% CL, 1.15, 1.67) after adjusting for baseline risk factors. CONCLUSIONS: The increased risk of CHD and death from all causes and cardiovascular diseases appeared to be only partially due to effects of smoking. No association was seen for lymphocytes or monocytes. PMID- 15780774 TI - Factors associated with errors in self-reports of stature, weight, and body mass index in Minnesota adolescents. AB - PURPOSE: Obtaining stature, weight, and body mass index (BMI) from self-reports rather than by direct measurements is highly attractive economically and logistically, but there are few data available for adolescents that allow evaluation of potential sources of reporting bias. Because self-reports are based on self perceptions, personal characteristics of youth may be related to errors in reporting. METHODS: Differences between self-reported and measured stature, weight, and BMI were investigated for a sample of 3797 Minnesota youth, 12 to 18 years of age. Gender, age, race/ethnicity, socioeconomic status (SES), and measured body size were examined as potential factors associated with errors in self-reports. RESULTS: Self-reported stature, weight, and BMI were generally highly correlated with corresponding measured dimensions, although adolescents of both genders systematically overestimated their statures, underestimated their weights, and underestimated BMI. Significant associations of errors in self reports with age, race/ethnicity, and socioeconomic status suggested that differences in self perceptions or value ascribed to body size varies according to these personal characteristics. Systematic errors in self-reported stature, weight, and BMI were negatively associated with the corresponding measured dimension when adjusted for age, race/ethnicity and SES, such that prevalences of overweight based on BMI from self-reported measures were systematically underestimated relative to measured values. CONCLUSIONS: Self-reports of stature, weight, and BMI are on the average, valid representations of their measured counterparts; nevertheless, errors in the self-reports are systematically related to characteristics of youth. Consequently, findings from these studies should be interpreted carefully. PMID- 15780775 TI - Mortality among participants in the agricultural health study. AB - PURPOSE: This analysis of the Agricultural Health Study cohort assesses the mortality experience of licensed pesticide applicators and their spouses. METHODS: This report is based on 52,393 private applicators (who are mostly farmers) and 32,345 spouses of farmers in Iowa and North Carolina. At enrollment, each pesticide applicator completed a 21-page enrollment questionnaire. Mortality assessment from enrollment (1994-1997) through 2000 provided an average follow-up of about 5.3 years, 447,154 person-years, and 2055 deaths. RESULTS: Compared with the general population in the two states, the cohort experienced a very low mortality rate. Standardized mortality ratios (SMRs) for total mortality, cardiovascular disease, diabetes, COPD, total cancer, and cancers of the esophagus, stomach, and lung were 0.6 or lower for both farmers and spouses. These deficits varied little by farm size, type of crops or livestock on the farm, years of handling pesticides, holding a non-farm job, or length of follow up. SMRs among ever smokers were not as low as among never smokers, but were still less than 1.0 for all smoking-related causes of death. No statistically significant excesses occurred, but slightly elevated SMRs, or those near 1.0, were noted for diseases that have been associated with farming in previous studies. CONCLUSIONS: Several factors may contribute to the low mortality observed in this population, including the healthy worker effect typically seen in cohorts of working populations (which may decline in future years), a short follow-up interval, and a healthier lifestyle manifested through lower cigarette use and an occupation that has traditionally required high levels of physical activity. PMID- 15780776 TI - Cigarette smoking and completed suicide among middle-aged men: a population-based cohort study in Japan. AB - PURPOSE: To clarify whether cigarette smoking was associated with completed suicide in a cohort of middle-aged Japanese men. METHODS: A total of 45,209 out of 57,714 men, aged 40 to 69 years, in nine public health centers across Japan responded to a self-administered questionnaire that included questions regarding their smoking history. The questionnaire was distributed in 1990 (Cohort I) and in 1993 and 1994 (Cohort II). By the end of 2000, 173 suicides were identified by death certificates. RESULTS: After adjusting for potential confounders, current smokers had a marginally higher risk of suicide than never smokers. Current smokers with more than 60 pack-years of cigarette smoking had a more than two fold higher risk of suicide than never smokers (pooled multivariate relative risk=2.1; 95% confidence interval, 1.1, 4.0), but a significant dose-response relationship between pack-years of smoking and suicide was not found. The risk of suicide increased significantly with increasing numbers of cigarettes smoked per day (p for trend=0.036). Smoking status, duration of quitting, duration of smoking, and age at the start of smoking were not significantly associated with suicide risk. CONCLUSION: Our results suggest that heavy smoking, particularly a large number of cigarettes per day at baseline was associated with an increased risk of suicide independent of several potential confounders. PMID- 15780777 TI - Regression models for clustered binary responses: implications of ignoring the intracluster correlation in an analysis of perinatal mortality in twin gestations. AB - PURPOSE: Dependent binary responses, such as health outcomes in twin pairs or siblings, frequently arise in perinatal epidemiologic research. This gives rise to correlated data, which must be taken into account during analysis to avoid erroneous statistical and biological inferences. METHODS: An analysis of perinatal mortality (fetal deaths plus deaths within the first 28 days) in twins in relation to cluster-varying (those that are unique to each fetus within a twin pregnancy such as birthweight) and cluster-constant (those that are identical for both twins within a sibship such as maternal smoking status) risk factors is presented. Marginal (ordinary logistic regression [OLR] and logistic regression using generalized estimating equations [GEE]) and cluster-specific (conditional and random-intercept logistic regression models) regression models are fit and their results contrasted. The United States "matched multiple data" file of twin births (1995-1997), which includes 285,226 twins from 142,613 pregnancies, was used to examine the implications of ignoring of clustering on regression inferences. RESULTS: The OLR models provide variance estimates for cluster constant covariates that ranged from 7% to 71% smaller than those from GEE-based models. This underestimation is even more pronounced for some cluster-varying covariates, ranging from 21% to 198%. CONCLUSIONS: Ignoring the cluster dependency is likely to affect the precision of covariate effects and consequently interpretation of results. With widespread availability of appropriate software, statistical methods for taking the intracluster dependency into account are easily implemented and necessary. PMID- 15780778 TI - Smoking cessation and lung cancer mortality in a cohort of middle-aged Canadian women. AB - PURPOSE: To determine the impact of smoking cessation on lung cancer mortality among women. METHODS: Survival analysis is used to assess the effect of smoking cessation on lung cancer death in the dietary cohort of 49,165 women aged 40 to 59 years enrolled in the Canadian National Breast Screening Study. RESULTS: During an average of 10.3 years of follow-up, 106 women died of lung cancer. The risk of lung cancer mortality among women who quit before age 50 (HR=0.26; 95% CI, 0.13-0.55 among women who quit at ages 40-49) or quit in the previous 10 years (HR=0.39; 95% CI, 0.22-0.69) is substantially lower than the risk among current smokers. Women who quit after age 40 or have quit for less than 20 years are at substantially higher risk of lung cancer mortality compared with never smokers. Both duration of smoking cessation and age at quitting have independent effects on lung cancer mortality, after controlling for number of cigarettes smoked per day and number of years smoked, as well as other potential confounding variables. CONCLUSION: These findings suggest that programs and policies to promote early cessation of smoking and prevention of relapse should be a public health priority. PMID- 15780779 TI - Health surveillance for asthma in the US Navy: experience of 9,185,484 person years. AB - PURPOSE: To measure trends and demographic risk factors for hospitalization for asthma. METHODS: Time trends and demographic risk factors, for hospitalized asthma (1CD-9-CM Code 493) were analyzed by measuring age-specific and age adjusted first hospitalization rates in a defined population of active-duty enlisted members of the US Navy worldwide during 1980-1999, consisting of 9,185,484 person-years. RESULTS: There were 3911 patients first hospitalized for asthma, including 2916 men and 995 women. The age-adjusted incidence rate of first hospitalization for asthma was three times higher in women than men, 110 per 100,000 person-years (95% confidence interval [CI], 104-117), compared with 35 per 100,000 person-years (95% CI, 33-37), respectively (p < 0.0001). The rate in black women was twice as high as in white women, 186 per 100,000 person-years, compared with 99 per 100,000 person-years, respectively (p < 0.001). The rate in black men was higher than in white men, 45 per 100,000, compared with 34 per 100,000 (p < 0.001). Age-adjusted rates in women doubled from 73 per 100,000 in 1980-1983 to 159 in 1997-1999 (p for trend < 0.01), while those in men remained stable. CONCLUSIONS: Age-adjusted incidence rates of first hospitalization for asthma were three times as high in women as in men, and doubled during the period between 1980 and 1999. The rates in black women were twice as high as in white women. The reasons are unknown. PMID- 15780780 TI - Depressive symptoms have an independent, gradient risk for coronary heart disease incidence in a random, population-based sample. AB - PURPOSE: Depression is a risk factor for incident coronary heart disease (CHD), and predicts poor prognosis for patients post-myocardial infarction (MI). Few population-based, prospective studies have tested a gradient risk for depressive symptoms on CHD incidence. METHODS: The sample (n=1302) was derived from the Nova Scotia Health Survey-1995 (NSHS95), an age- and sex-stratified, random, population-based health survey. All subjects were 45 years or older, free of overt CHD at baseline, and completed the Center for Epidemiological Studies Depression (CES-D) scale. Covariates included age, sex, body mass index, physical activity level, family history of premature CHD, diastolic blood pressure, lipids, smoking, alcohol use, diabetes, and education level. For the 4 years following NSHS95, MI-related hospitalizations (ICD-9-CM code 410) and CHD-related deaths (ICD-9-CM codes 410-414) were extracted from the provincial, universal healthcare registry. RESULTS: Fifty-two participants experienced a CHD event. A one standard-deviation increase in CES-D score was associated with a 1.32 hazard risk (confidence interval, 1.01-1.71) of CHD events, controlling for established CHD risk factors. CONCLUSIONS: An independent, gradient association between depression and incident CHD was detected in a population-based sample with complete 4-year CHD data. This evidence supports the value of investigating mechanisms linking depression and CHD. PMID- 15780781 TI - The limits of abstinence-only in preventing sexually transmitted infections. PMID- 15780782 TI - After the promise: the STD consequences of adolescent virginity pledges. AB - PURPOSE: To examine the effectiveness of virginity pledges in reducing STD infection rates among young adults (ages 18-24). METHODS: Data are drawn from the National Longitudinal Study of Adolescent Health, a nationally representative study of students enrolled in grades 7-12 in 1995. During a follow-up survey in 2001-2002, respondents provided urine samples, which were tested for Human Papilloma Virus, Chlamydia, Gonorrhea, and Trichomoniasis. We report descriptive results for the relationship of pledge status and sexually transmitted disease (STD) rates as well as health behaviors commonly associated with STD infection. RESULTS: Pledgers are consistently less likely to be exposed to risk factors across a wide range of indicators, but their STD infection rate does not differ from nonpledgers. Possible explanations are that pledgers are less likely than others to use condoms at sexual debut and to be tested and diagnosed with STDs. CONCLUSIONS: Adopting virginity pledges as intervention may not be the optimal approach to preventing STD acquisition among young adults. PMID- 15780783 TI - Religious differentials in the sexual and reproductive behaviors of young women in the United States. AB - PURPOSE: To examine whether religious involvement and affiliation during early adolescence is associated with reduced levels of sexual risk. METHODS: We used nationally representative data from the 1995 National Survey of Family Growth (NSFG) to examine the relationship between religious affiliation and frequency of attendance at religious services at age 14 years and a range of sexual behaviors among women aged 15-24 years, including age of first intercourse, contraceptive use, timing of first family planning visit, timing of birth, and number of sexual partners. The majority of young women had mothers with at least a high school education (79%) and had been living with both parents at age 14 (57%). Two-thirds of the sample was white, 12% Hispanic, and 15% black, and a substantial minority is poor (18%) or low-income (24%). We used bivariate analysis to describe variation in behavior according to religious affiliation and frequency of attendance. We used multivariate techniques to assess the role of religion once other factors are taken into account. RESULTS: Bivariate relationships suggest that both affiliation and attendance are associated with age of first sex, contraceptive use at first sex, teen births, and number of sexual partners. However, most of these associations disappear once we control for young women's demographic characteristics. Multivariate analyses show that religious affiliation shares few associations with sexual behaviors, though frequent attendance at religious services at age 14 years continues to have a strong delaying effect on the timing of first intercourse. CONCLUSION: Frequency of attendance and religious affiliation have little impact on sexual behaviors once intercourse occurs. PMID- 15780784 TI - The impact of life skills education on adolescent sexual risk behaviors in KwaZulu-Natal, South Africa. AB - PURPOSE: To assess the impact of exposure to life skills education by youth in KwaZulu-Natal Province (KZN, South Africa) on knowledge and behaviors associated with the spread of HIV/AIDS. METHODS: Data come from a panel study of 2222 youth from several population subgroups in KZN. The youth were aged 14-24 years when interviewed in 1999 and 2001. The intervention was a full coverage life skills education program for all students in middle and secondary schools. Impact was measured as net dose-response relationships between life skills exposure and outcomes. Econometric methods were used to control for nonrandom exposure to life skills education. Outcomes included sexual behavior and condom use indicators. RESULTS: Significant effects, albeit modest in magnitude, were observed on selected areas of sexual-reproductive health knowledge and perceived condom self efficacy, along with larger effects on condom use at first and last sex. No consistent effects on age at sexual initiation, secondary abstinence, or partnering behaviors were observed among these youth. CONCLUSIONS: School-based life skills education appears capable of communicating key information and helping youth develop skills relevant to reducing HIV risk. However, the South African national program has yet to be fully implemented, and whether this initiative will result in sustained behavior modification among youth on a sufficient scale to affect the HIV/AIDS epidemic is uncertain. PMID- 15780785 TI - Osteoporosis knowledge, beliefs, and practices among adolescent females. AB - PURPOSE: To assess knowledge, beliefs, and practices of osteoporosis risk factors (physical activity, smoking, and nutrition) among adolescent girls. METHODS: Questionnaires consisting of items assessing knowledge, beliefs, and practices regarding physical activity, calcium intake, and cigarette smoking were administered to 227 girls aged 12-16 years. The adolescent girls were attending public and separate schools in a small-sized Canadian city. Descriptive statistics, t-tests, and Chi-square tests were used to analyze the data. RESULTS: Female adolescents believed that physical inactivity, smoking, and inadequate calcium were health-risking behaviors and osteoporosis risk factors, however, specific in-depth knowledge regarding these risk factors was lacking. Findings further showed that health-risking behaviors were evident, as 25% were current smokers, 58% consumed less than the adequate intake for calcium, and 52% had scores that reflected low to moderate physical activity levels. CONCLUSION: These results highlight the need for educational interventions to promote specific osteo-protective practices among adolescent girls. PMID- 15780786 TI - Exposure to community violence and upper respiratory illness in older adolescents. AB - PURPOSE: To examine the relationship between exposure to chronic community violence and upper respiratory illness (URI) symptoms among urban adolescents of color; and to test the generality of a model of the relationship between social stress and URI. METHOD: The research used a cross-sectional correlational design. The sample was 769 first-semester first-year students in an urban nonresidential 4-year college from the academic years 1999-2002. Data were collected by a group administered questionnaire in academic classes. The research used three multi item additive scales (each with high reliability and validity): exposure to community violence, psychological distress, and URI symptoms. Multiple regression procedures were used to analyze the data. RESULTS: Positive correlations were obtained between: exposure to community violence and reporting of URI (r = .19), exposure and psychological distress (r = .22), and psychological distress and URI (r = .51). The relationship between exposure to community violence and URI is greatly reduced when level of psychological distress is statistically controlled. CONCLUSIONS: Exposure to community violence is related to experiencing URI symptoms among older urban adolescents of color; the effect size of the relationship is small-medium. Psychological distress mediates the impact of exposure to community violence on URI. The findings expand the range of social stressors that are empirically related to URI, and populations in which a relationship between social stressors and URI may be found. PMID- 15780787 TI - Weight management goals and use of exercise for weight control among U.S. high school students, 1991-2001. AB - PURPOSE: The purpose of this study was to examine trends in weight management goals and the use of exercise for weight control among U.S. high school students. METHODS: As part of the Youth Risk Behavior Surveillance System, 6 national school-based surveys were conducted between 1991 and 2001. Each survey used a three-stage cross-sectional sample of students in grades 9-12. African-American and Hispanic students were oversampled. Logistic regression models were used to test for trends among gender and race/ethnic subgroups, controlling demographic changes over time. RESULTS: From 1991 to 2001, the percentage of female students trying to lose weight (61.7%-62.3%) or stay the same weight (15.4%-16.0%) did not change significantly. Among male students, trying to lose weight (22.7%-28.8%) and trying to stay the same weight (17.8%-21.5%) both increased significantly, while trying to gain weight decreased significantly (32.7%-26.3%). Among female and male students who were trying to lose weight or stay the same weight, the use of exercise for weight control increased significantly. Among students who reported using exercise for weight control, participation in vigorous physical activity > or = 3 days per week increased among African-American female students, and participation in strengthening exercises > or = 3 days per week increased among male students. CONCLUSIONS: These findings suggest increased interest in weight control among male adolescents, and increased use of exercise for weight control among female and male adolescents. PMID- 15780788 TI - Reliability and validity of the VSP-A, a health-related quality of life instrument for ill and healthy adolescents. AB - PURPOSE: To report the preliminary validation results of a generic self administered measure for adolescents the VSP-A (Vecu et Sante Percue de l'Adolescent). METHODS: The validation survey of the 37-item VSP-A involved 1938 adolescents: 1758 adolescents attending school (90.7%) and 180 inpatient youths (9.3%) with medical, surgical, or psychiatric conditions; mean age was 14.8 years (SD = 2.1; range = 10-17) and 1018 (52.5%) were girls. Ten dimensions scores were computed: psychological well-being, body image, physical well-being, vitality, friends, parents, teachers, school performance, medical staff, and a global HRQL index. Construct validity was tested using inter-item correlations, item dimension correlations, and principal component analysis. The unidimensionality of the scales was studied using Rasch rating scale model and the internal consistency of the scales using Cronbach alpha. Known group comparisons were performed and effect sizes computed to assess the clinical validity and sensitivity of the instrument. RESULTS: VSP-A, completed in less than 15 minutes, is consistent (Cronbach alpha = 0.74-0.91). Content and construct validity are good. Factorial validity, item-internal consistency, item-discriminant validity, and inter-dimension correlations support the item grouping in 10 dimensions. The results of the comparison of the VSP-A scores according to external criteria (sociodemographics, health status) show they are in accordance with the literature and previous works. CONCLUSIONS: These preliminary results support the reliability and validity of the VSP-A as a multidimensional adolescent report of HRQL in healthy and patient populations. The next phase of the research consists of testing the sensitivity to changes of the VSP-A. PMID- 15780789 TI - Performance of urine leukocyte esterase in asymptomatic male youth: another look with nucleic acid amplification testing as the gold standard for Chlamydia detection. AB - PURPOSE: To re-evaluate the sensitivity and specificity of leukocyte esterase (LE) for screening adolescent and young adult males for Chlamydia trachomatis using a nucleic acid amplification test (NAAT) as the gold standard. METHODS: This study was conducted at two Massachusetts Department of Youth Services sites and one Job Corps site. Recently admitted asymptomatic sexually active male youth aged 14 to 25 years (mean 16.6 years) were recruited between January 2001 and July 2003 (N = 1008). Participants provided first part voided urine specimens for testing with LE and Chlamydia NAAT. The sensitivity, specificity, and positive and negative predictive value of urine LE for identification of Chlamydia infection were determined using NAAT as the gold standard. RESULTS: Fifty-seven (5.7%) participants were infected with Chlamydia as defined by a positive NAAT. Defining trace + as the LE cut point resulted in sensitivity and specificity of 57.9% and 78.3%, respectively. Defining 1+ as the cut point resulted in sensitivity and specificity of 47.4% and 96.1%, respectively. CONCLUSIONS: Urine leukocyte esterase is a moderately sensitive method to screen for Chlamydia. Nevertheless, a substantial proportion of infections are not detected with LE screening. When feasible, urine NAAT provides a much more sensitive and equally noninvasive method of detecting Chlamydia. However, if LE is used as an initial screen followed by NAAT confirmation of LE positive samples, we recommend using trace LE as the cut point for positive results. PMID- 15780790 TI - Behavioral risk in early adolescents with HIV+ mothers. AB - PURPOSE: To examine the effect of maternal human immunodeficiency virus (HIV) infection on sexual and drug use risk behavior, delinquency, and general behavior problems in early adolescents. METHODS: Baseline data from an ongoing longitudinal study are presented. Participants are 220 HIV-negative early adolescents (aged 10-14 years), 100 with HIV-infected mothers, and 120 with uninfected mothers from ethnic minority, low income, families living in inner city communities. For two group comparisons, Chi-square and Student's t-tests were used. Multiple and Logistic regressions were conducted to control for age and examine multiple predictors simultaneously. RESULTS: Few early adolescents, particularly those younger than 13 years, reported penetrative sexual behavior (oral, anal, or vaginal sex, 7%) or drug use (12%). Nonpenetrative sexual behaviors (kissing, 35%), alcohol use (39%), and engaging in at least one delinquent activity (45% of boys, 26% of girls) were more common. By age 14 years, 21% of the youths reported penetrative sexual behavior and 72% reported alcohol use. Penetrative sexual behavior was significantly associated with delinquent behavior and substance use. There were no differences in risk behaviors between youth with and without HIV-infected mothers. However, among youth with HIV+ mothers, those who knew their mother's status had more thought problems (p = .042) and reported more frequent alcohol use (p = .018) than those youth who didn't know. CONCLUSIONS: Maternal HIV status did not significantly add to the risk for problem behaviors in our sample of urban ethnic minority early adolescents. Fourteen years of age appeared to be a critical time for increased experimenting with sexual behavior and substance use. PMID- 15780791 TI - Body piercing practices and attitudes among urban adolescents. AB - PURPOSE: This article describes adolescents' attitudes and practices toward body piercing and their awareness of associated health risks. METHODS: An anonymous, 32-item, random, convenience-sample survey was administered to 225 participants, ages 12-21 years, at an urban, hospital-based, adolescent clinic. RESULTS: The mean participant age was 16.5 +/- 2.1 years; 78% were female; and 58% were African-American and 30% were Caucasian. Forty-eight percent had a body piercing. About half of participants (range, 45%-62%) endorsed each site as accepted by the public, except for nipple and genital sites, which fewer than 10% endorsed as accepted. African-Americans were more likely to have pierced noses, and to find that site acceptable, whereas Caucasians were more likely to have pierced navels and nipples, and to find those sites acceptable (odds ratio [OR] range, 3.05 19.37). Youth tended to see their own pierced site as more acceptable and less risky than others did (OR range, 8.99-23.61; effect sizes [r] range, .15-.60). The most common health problems from body piercing reported by participants from their personal experience were infection (10%), allergic reaction (1%), and bruising (1%). The most common health problems from body piercing reported by participants based on others' experiences were infection (74%), bleeding (30%), allergic reaction (26%), bruising (19%), and keloids (19%). Those who were pierced in a body-piercing shop reported more infections (18.4%) than those pierced in other places (1.9%) (OR, 11.49; 95% confidence interval [CI], 1.39 90.91). Youth who had pierced themselves perceived less risk from piercing from a nonprofessional (50%) compared with those pierced by a professional (77%) (p = .003). CONCLUSIONS: Youth from this hospital-based urban adolescent clinic considered body piercings to be generally accepted by the public. Pierced youth in our survey often participated in body piercing practices that they perceived as minimally risky and that they defined as normative. Based on these findings, clinicians should discuss body piercing in the context of providing routine anticipatory guidance to adolescents and should educate adolescents about safer piercing strategies to help minimize associated health risks. PMID- 15780792 TI - Suicidal behavior in the family and adolescent risk behavior. AB - PURPOSE: The purpose of this study was to examine the prevalence and sociodemographic characteristics of adolescents exposed to suicide attempts and suicide deaths by a family member and to evaluate the separate associations between exposure to a family member's suicide attempts or suicide death and risk behaviors, social-emotional functioning, and family connectedness. METHOD: We performed a cross-sectional analysis of data collected in 1995 for wave I of the National Longitudinal Survey of Adolescent Health (n = 5,918). The independent variable was the exposure to suicidal behavior by a family member during the past 12 months (death by suicide, suicide attempt, and no suicidal behavior). The dependent variables were substance use, suicidal behavior, violent behavior, perceived shortened life expectancy, emotional distress, and parental and adolescent reports of family connectedness. RESULTS: In the year before the survey, 3.9% of adolescents experienced a family member's suicide attempt whereas 1.2% experienced a family member's death by suicide. In regression analyses adjusting for sociodemographic variables, adolescents who had experienced the suicide attempt of a family member were more likely than those with no exposure to report the following: cigarette and marijuana use, alcohol misuse, suicidal ideation and attempts, fighting and inflicting injuries, decreased life expectancy, emotional distress, and decreased adolescent reports of parent-child and family connectedness. Adolescents who had experienced a family member's death by suicide were more likely to report marijuana use and alcohol misuse, suicidal ideation and attempts, inflicting severe injuries, and emotional distress. CONCLUSIONS: Adolescents who have experienced suicide attempts or suicide deaths in the family show high levels of at-risk behaviors, most notably their own suicidal ideation and attempts. PMID- 15780794 TI - Preadmission patterns of physical activity in a sample of juvenile detainees. AB - Preadmission patterns of physical activity were examined in a sample of 537 juvenile detainees and compared with the 2001 Youth Risk Behavior Survey (YRBS), a national sample of high school students. Overall, detained youth reported similar participation in physical activity, surpassing the student sample in some areas. PMID- 15780793 TI - Are pregnant adolescents stigmatized by pregnancy? AB - PURPOSE: To identify prevalence and correlates, including substance use and exposure to violence, of feeling stigmatized by being pregnant as an adolescent. METHODS: A total of 925 low-income African-American, Mexican-American, and Caucasian pregnant adolescents aged C substitution at 94 was found only among the IDUs. CONCLUSIONS: These observations suggest that the current HCV transmission between the IDU and non-IDU populations is uncommon, and may reflect the beginning of divergence of HCV genotypes in the IDU population in Prague. PMID- 15780810 TI - Circulating human antibodies against dengue NS1 protein: potential of recombinant D2V-NS1 proteins in diagnostic tests. AB - The dengue virus (DV) causes one of the most important arthropod-borne human viral diseases throughout the tropical and subtropical countries. However, the morbidity and mortality of DV infections could be reduced with an early hospitalization care and a rapid risk identification of developing the dengue haemorrhagic fever (DHF). The nonstructural glycoprotein 1 (NS1) has been pointed as a reagent for immune-assay diagnostic test optimization. To evaluate this potential, recombinant DV2-NS1 proteins (rNS1) were produced from Escherichia coli (NS1EC) and insect cells (NS1IC) expression. The tests were performed by analysis of a human serum panel reacted against different rNS1 forms. The results demonstrated high correspondence between the DV positive sera and the assay results using native or refolded forms of either NS1IC or NS1EC. Also, the IgG and IgM anti-rNS1 level profiles showed distinct distribution, depending on protein form and disease status. However, the IgM anti-rNS1 reactions did not show sensibility to detect the DV in primary infections. The data obtained from the paired serum samples reactivity comparison suggested a heterogeneous human immune response and absence of correspondence between the IgG and IgM profile levels. Moreover, a patient with negative reference test could be detected by specific IgG anti-rNS1 assays presented here. Therefore, these results sustain the usefulness of dengue nonstructural proteins, in particular the NS1, in diagnostic tests as a complementary reagent. PMID- 15780811 TI - Development of a semi-quantitative real-time RT-PCR for the detection of measles virus. AB - Real-time detection of polymerase chain reactions allows convenient detection and quantification of virus-derived nucleic acids in clinical specimens. We have developed a real-time RT-PCR assay for the detection of measles virus (MV) genomic RNA, and compared it to a well-established conventional RT-PCR assay. Based on a serial dilution of the live-attenuated MV Edmonston Zagreb vaccine, the detection limits were approximately 0.1 and 0.02 cell culture infectious dose 50% units (CCID50) per test for the conventional and TaqMan RT-PCR assays, respectively. Furthermore, tissue materials spiked with known quantities of MV were equally well detected in both assays. The TaqMan assay was linear within a range of 10(4.4) to 10(-0.6)CCID50/ml, with an intra-assay variability lower than 3% and an inter-assay variability ranging from 1.5% at 10(4.4)CCID50/ml to 8.7% at 10(-0.6)CCID50/ml. The TaqMan assay could detect representative wild-type viruses from the currently active MV clades, and could detect MV genome in clinical specimens obtained from measles patients. Finally, quantification of MV RNA in peripheral blood mononuclear cells or broncho-alveolar lavage cells from cynomolgus macaques collected at different time points after experimental infection showed a good correlation with virus isolation data. In conclusion, the TaqMan assay developed is specific, sensitive, rapid and reproducible, and can be of use for diagnostic purposes or for studies on the pathogenesis of measles. PMID- 15780812 TI - Development of a multiplex nested consensus PCR for detection and identification of major human herpesviruses in CNS infections. AB - BACKGROUND: Rapid, sensitive and economical detection and identification of human herpesviruses as causative agents of central nervous system (CNS) infections are of clinical importance. The traditional methods for the detection of herpesviruses in CNS infections all suffer from limitations. PCR has a potential to overcome each of them. OBJECTIVES: The aims of this study were reducing the number of primers in multiplex PCR and increasing the sensitivity of the assay by nested PCR. STUDY DESIGN: A multiplex nested consensus PCR (MNC-PCR) was developed for the simultaneous detection of major human herpesviruses. A pair of conserved primers was designed for detection of HSV-1, HSV-2, CMV and EBV and another pair of conserved primers for nested PCR. For VZV, a different pair of primers was designed and another pair of primers for nested PCR. A reduction in the number of designed primer pairs (from five pairs to two in both stages of PCR) is an advantage in this assay. One hundred forty-seven cerebral spinal fluid (CSF) samples from patients that showed clinical manifestation of CNS infections were tested. Results of MNC-PCR in CSF samples were compared with those of single PCR assay for each individual DNA virus. Sensitivity of the assay was determined with a plasmid containing VZV DNA binding protein gene and another plasmid for HSV-1 DNA polymerase gene. False negative results (due to the presence of inhibitor of DNA amplification in CSF samples) were avoided by the inclusion of beta2-microglobulin primers in the MNC-PCR assay as an internal control. RESULTS: Positive results were obtained in 20 CSF samples (8 HSV-1, 2 HSV-2, 4 CMV, 3 VZV, 3 HSV-1/CMV, CMV/VZV and HSV-1/EBV coinfections). The comparison between single PCR and MNC-PCR showed a marked increase in sensitivity of MNC-PCR test, since six negative samples in single PCR proved positive in MNC-PCR (P<0.005). Sensitivity was determined 1-5 plasmid copies for VZV and 50-100 plasmid copies for HSV-1. CONCLUSIONS: The MNC-PCR assay presented in this study can provide a rapid, sensitive and economical method for detection of viral infections and is applicable to small volumes of CSF samples. PMID- 15780813 TI - Detection of unusual strains of RV in patients with acute diarrhoea in Mexico. AB - BACKGROUND: Group A rotaviruses are a major cause of acute gastroenteritis in infants. Human strains with a short RNA pattern generally exhibit subgroup I, G2, P1B[4] specificity, those with a long RNA pattern show subgroup II, G1, G3 or G4, P1A[8] specificity. The presence of strains with unusual specificities has been reported worldwide over the last decade. OBJECTIVE: To determine antigenic diversity among rotaviruses isolated from patients with diarrhoea. STUDY DESIGN: A laboratory-based survey study was carried out with faecal samples from patients with acute gastroenteritis form January to April 2000. To classify the samples PAGE and ELISA with specific antibodies to serotype G and P and RT-PCR were carried out. RESULTS: Twenty one specimens from patients with dehydrating diarrhoea had unusual specifies. Nine specimens had unusual combination of long pattern and subgroup I. Twelve specimens with short pattern belong to G1 serotype. As far as the serotypes and genotypes concern 11 samples were P1A, P[4] and one specimen was P1A, P[9]. CONCLUSIONS: These results demonstrated the unexpected presence of unusual strains of rotavirus in Mexico. Detection of strains with both human and animal characteristics may indicate interspecies transmission of RV between humans and animals. PMID- 15780814 TI - Apoptosis as a mechanism of natural resistance to HIV-1 infection in an exposed but uninfected population. AB - BACKGROUND: Apoptosis, also known as programmed cell death, has been reported not only as a pathogenic mechanism, but also as a mechanism of resistance and control of a variety of infections. Particularly during HIV-1 infection, apoptosis is the main mechanism by which infected and uninfected CD4+ lymphocytes are eliminated. However, apoptosis as a mechanism of natural resistance to HIV infection has this far not been explored. OBJECTIVE: To determine whether apoptosis could explain, at least in part, the natural resistance to HIV infection observed in some exposed but uninfected individuals (ESN). RESULTS: Our data shows that peripheral blood monocytes in the ESN group has a predisposition to undergo spontaneous apoptosis, as well as apoptosis induced by HIV infection in vitro, compared with monocyte population from the control group at low risk of HIV infection. CONCLUSIONS: These findings suggest that, in some ESN individuals, monocytes could play an important role in the control of HIV infection by undergoing apoptosis. However, since the variability among individuals is large, studies with larger cohorts focusing in monocyte apoptosis as pathogenic mechanisms are required. PMID- 15780816 TI - Influence of HTLV-2 infection on hepatitis C virus replication in HIV-positive patients. PMID- 15780815 TI - Prevalence of ARV resistance mutations and impact of genotyping test in HIV patients with advance disease in Sao Paulo, Brazil. PMID- 15780817 TI - Quantitative cytomegalovirus load in urine and clinical outcome in neonates. PMID- 15780819 TI - Adipose tissue as an active endocrine organ: recent advances. AB - Adipose tissue secretes a variety of factors in a manner dependent upon its metabolic state. These factors are derived from adipocyte or non-adipocyte fractions, and include proteins, metabolites and hormones. Obesity is a major risk factor for type 2 diabetes and cardiovascular disease, and adipocyte-derived factors might contribute to or ameliorate obesity-associated pathologies such as insulin resistance, dyslipidemia, vascular dysfunction and a chronic inflammatory and prothrombotic state. PMID- 15780820 TI - The role of the adipocyte hormone adiponectin in cardiovascular disease. AB - Adiponectin, a novel hormone made by fat tissue, regulates energy metabolism and endothelial activation. Serum levels of adiponectin are reduced in conditions that are associated with an increased risk of cardiovascular disease, such as diabetes and the metabolic syndrome. Adiponectin trimers assemble into higher order oligomers, which have different signaling properties. Adiponectin trimers and a C-terminal globular domain activate AMP-activated protein kinase, whereas hexamer and high-molecular weight isoforms activate nuclear factor-kappa B signaling pathways. Exogenous adiponectin corrects metabolic defects that are associated with insulin resistance, and might protect the endothelium from the progression of cardiovascular disease. Receptors for adiponectin have been described and might provide future therapeutic targets for the treatment of cardiovascular disease. PMID- 15780821 TI - Metabolic syndrome-interdependence of the cardiovascular and metabolic pathways. AB - The metabolic syndrome is a worldwide epidemic, setting the stage for type 2 diabetes and its microvascular complications, and acceleration of macrovascular disease. Insulin resistance, hyperglycemia, dyslipidemia, hypertension, thrombotic disorders and adiposity define the metabolic syndrome and contribute to endothelial dysfunction and, subsequently, to accelerated atherosclerosis. Angiotensin II contributes to the development and progression of cardiovascular and renal endpoints and, as such, angiotensin II receptor blockers and angiotensin-converting enzyme inhibitors demonstrate a protective effect. Ligands for the peroxisome proliferator-activated receptor gamma (PPAR gamma), appear to impact favourably on atherosclerosis through both direct and indirect mechanisms. In humans, these ligands improve endothelial function, attenuate albuminuria and hypertension, and potentially prevent conversion of prediabetes to type 2 diabetes. Statins also have proven benefit in decreasing overall cardiovascular and stroke mortality and morbidity. The combination of angiotensin II blockade, statin therapy and PPAR gamma activation might emerge as an important global therapeutic strategy in the metabolic syndrome and diabetes. Further studies are needed to determine whether they have synergistic effects to protect the vasculature. PMID- 15780822 TI - Cardiovascular complications in diabetes mellitus. AB - Diabetes is strongly associated with coronary, cerebral and peripheral arterial disease, as well as with microangiopathy. In those with diabetes, the extent of macrovascular disease increases and atherosclerotic plaques are more prone to rupture. Both hormonal abnormalities (insulin resistance that is typically present for many years before the onset of type 2 diabetes) and metabolic abnormalities contribute. Multi-targeted intensive therapy is imperative; however, unfortunately it is underutilized. Functional and structural derangements contribute to impaired arterial and ventricular compliance predisposing to congestive heart failure that is increasingly recognized to be a cause of morbidity and mortality in patients with diabetes. PMID- 15780823 TI - Plasminogen activator inhibitor-1: a common denominator in obesity, diabetes and cardiovascular disease. AB - A classical perspective of cardiovascular risk does not adequately account for all of the cardiovascular events associated with obesity and diabetes. The combination of hypertriglyceridemia, glucose intolerance and inflammation is linked with increased production of the primary inhibitor of endogenous thrombolysis, plasminogen activator inhibitor-1 (PAI-1). Recent data suggest that PAI-1 contributes directly to the complications of obesity, including type 2 diabetes, coronary arterial thrombi, and may even influence the accumulation of visceral fat. Therefore, direct inhibition of PAI-1 might not only provide a new therapeutic strategy for reducing cardiovascular risk, but may also have beneficial effects on obesity and insulin resistance. PMID- 15780824 TI - Hypercoagulability in the metabolic syndrome. AB - The metabolic syndrome is characterized by a combination of obesity, chronic inflammation and insulin resistance. This syndrome also has features of a hypercoagulable state, consisting of increased levels of clotting factors (tissue factor, factor VII and fibrinogen) as well as inhibition of the fibrinolytic pathway (increased plasminogen activator inhibitor-1 and decreased tissue plasminogen activator activity). Simultaneously, the presence of endothelial dysfunction and dyslipidemia triggers platelet aggregability, thus further increasing the risk of thrombotic events both in the arterial and venous system. Although mechanisms of coagulation activation are well described for other diseases, the precise etiology is not well known in the metabolic syndrome. Thus far, only obesity has been shown to be a modest risk factor for venous thromboembolic events, whereas accurate data for metabolic syndrome patients are lacking. Hence, routine interventions for prevention of venous thromboembolism are not yet warranted. However, as dyslipidemia is associated with procoagulant changes, this could be a possible target for therapeutic intervention. In view of the rising incidence of metabolic syndrome even at a young age, both the incidence of venous thromboembolism and the effect of intervention on markers of hypercoagulability in metabolic syndrome call for further studies. PMID- 15780825 TI - Insulin resistance, hyperleptinemia and endothelial dysfunction in coronary restenosis. AB - Impaired insulin sensitivity and endothelial dysfunction are important markers in the development of restenosis after coronary stenting. In addition, new markers of inflammation and endothelium activation, such as increased leptin levels, also have to considered. Many studies have shown that hyperinsulinemia and insulin resistance increase neointimal index measured six months after coronary stenting, and that insulin-sensitizers have beneficial effects by decreasing the rate of restenosis. The role of endothelial dysfunction in the process of restenosis is a fascinating problem. The pathobiology of restenosis in stented arteries is largely related to neointimal hyperplasia, which is dependent upon several factors, such as a reduction in nitric oxide activity that determines endothelial dysfunction and oxidative stress. Abnormal endothelium-dependent vasodilation (related to decreased nitric oxide production in the insulin-resistant state) might be explained by alterations in intracellular signaling and increased endothelin-1 production. Leptin is a hormone related to both fat metabolism and insulin resistance that has been recognized as an independent predictor of coronary restenosis. Chronic hyperleptinemia can reduce the synthesis of nitric oxide owing to the increased oxidative stress in endothelial cells. As a result, the goal in prevention of in-stent restenosis is to develop drugs that are able to act both as insulin- and endothelium-sensitizers. PMID- 15780826 TI - Targeting survival signaling in heart failure. AB - Accumulating evidence suggests that apoptosis is not only a common feature of diverse forms of heart failure but also contributes to disease pathogenesis and progression. This contribution of apoptotic signaling to heart failure could reflect not only loss of cardiomyocytes but also dysfunction of surviving cells. The convergence of signaling mechanisms controlling both cardiomyocyte survival and function provides an opportunity for therapeutic strategies that target these pathways. However, significant hurdles must be overcome before the clinical application of these insights becomes possible. PMID- 15780828 TI - Role of the PPAR family of nuclear receptors in the regulation of metabolic and cardiovascular homeostasis: new approaches to therapy. AB - Several conditions and risk factors predispose to cardiovascular disease, including visceral obesity, type 2 diabetes, insulin resistance, hypertension and dyslipidemia, and are collectively called the metabolic syndrome. Different pharmacological approaches are under development to address these risk factors and improve the cardiovascular risk profile. Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors that control lipid and glucose metabolism, as well as the inflammatory response. Thus PPARs appear good drug targets for the correction of the global risk profile that predisposes an individual to cardiovascular disease. PMID- 15780827 TI - Nuclear receptors and their relevance to diseases related to lipid metabolism. AB - Drugs that target the nuclear hormone receptor family constitute one of the largest and most potent groups of pharmaceuticals currently in use. However, although many of these human nuclear receptors have been clearly demonstrated to be key sensors and regulators of lipid metabolism, the full pharmacological potential of this drug target class has not been fully explored. There are two main reasons for this. First, a rationale approach is needed to identify pharmacologically selective drug candidates to nuclear receptors that have a large therapeutic window between the beneficial effects and the unwanted side effects. This appears to apply to all ligand-regulated nuclear receptors, including those nuclear receptors more recently proposed as novel targets for diseases related to lipid metabolism such as the peroxisome proliferator activated receptors, liver X receptors and farnesoid X-activated receptor. The second reason is that any sub-group of nuclear receptors important for the regulation of lipid metabolism might be pharmacologically inaccessible by conventional low molecular weight compounds, owing to the lack of a classical ligand-binding-pocket, as recently revealed by X-ray crystallography. Accordingly, targeting of classical nuclear receptor family members with better characterized endocrinology and roles in lipid metabolism, such as the thyroid and steroid hormone receptors, could become of renewed pharmacological interest, as these targets provide well-characterized alternatives to the more recently discovered nuclear receptors. PMID- 15780829 TI - Acarbose in the prevention of cardiovascular disease in subjects with impaired glucose tolerance and type 2 diabetes mellitus. AB - The prevalence of glucose intolerance is increasing dramatically worldwide. Both impaired glucose tolerance (IGT) and type 2 diabetes are associated with excess mortality from cardiovascular disease. It is now generally accepted that these cardiovascular complications are related to prevailing hyperglycemia, particularly postprandial hyperglycemia. Acarbose specifically decreases the postprandial glycemic surge in IGT and diabetic subjects. The Study To Prevent Non-insulin-dependent Diabetes Mellitus (STOP-NIDDM) trial has shown that acarbose treatment in IGT subjects decreased the risk of progression to diabetes by 36%. Furthermore, it was associated with a 49% risk reduction of cardiovascular events. In a subgroup of subjects, acarbose treatment was accompanied by a 50% decrease in the progression of intima-media thickness of the carotids. Finally, a meta-analysis of seven major studies on the use of acarbose in the treatment of diabetes indicated that acarbose treatment was associated with a 35% risk reduction of cardiovascular disease. It is proposed that the mechanism by which acarbose can lower the risk of cardiovascular events is through diminution of oxidative stress induced by postprandial glycemic excursion. PMID- 15780830 TI - Aspirin response and failure in diabetic patients with cardiovascular disease. AB - Platelets are known to play a pivotal role not only in the formation of arterial thrombosis but also in the progression of atherosclerotic disease. Recent data suggest that these activities of platelets have become enhanced in patients with type 2 diabetes, accounting for many of the cardiovascular complications. Although aspirin is the most commonly used drug to regulate platelet function, many patients do not receive the expected clinical benefit of this drug and do not have the expected inhibition of platelet function, perhaps because of platelet hyperactivity in diabetes. The data suggest that aspirin might be a less than adequate antithrombotic for type 2 diabetes and that alternative therapies (e.g. combinations with clopidogrel or new antithrombotic therapies) are required to effectively manage the prevalent thrombotic complications associated with this disease. PMID- 15780831 TI - Isoprostane formation and inhibition in atherothrombosis. AB - Enhanced oxidant stress plays a pivotal role in the pathogenesis of several conditions such as atherothrombosis, cancer and neurodegeneration. The availability of reliable assays of isoprostanes in biological fluids has prompted clinical investigations into the pathophysiological role of lipid peroxidation in cardiovascular disease. Established risk factors for coronary heart disease have been associated with enhanced oxidant stress, leading to increased lipid peroxidation and non-enzymatic formation of bioactive isoprostanes. These risk factors include diabetes mellitus, hypercholesterolemia, obesity and hyperhomocysteinemia. Clinical studies support the hypothesis that enhanced lipid peroxidation may contribute, at least in part, to persistent platelet activation. PMID- 15780833 TI - The Ixodes scapularis Genome Project: an opportunity for advancing tick research. AB - The Ixodes scapularis Genome Project (IGP), the first to sequence a tick genome, will provide an unparalleled resource for studying tick biology and tick-host pathogen relationships, and identifying novel targets for tick and tick-borne disease control. The IGP will be the first genomic analysis of a member of the subphylum Chelicerata and will accelerate the pace of tick research. The challenge for scientists is to translate IGP data into public health benefits. PMID- 15780832 TI - Cardiovascular hazard and non-steroidal anti-inflammatory drugs. AB - Selective inhibitors of cyclooxygenase (COX)-2 depress prostacyclin (PGI(2)) without a concomitant inhibition of platelet COX-1-derived thromboxane (Tx)A(2). Experiments in gene-deleted mice have shown that ablation of the PGI(2) receptor (the IP) predisposes to an exaggerated response to agonists which elevate blood pressure, accelerate atherogenesis and induce thrombosis. Such a class-based effect would be expected to be modulated by the underlying risk of cardiovascular disease in patients, elements of drug exposure, such as dose, duration of action and duration of dosing, and inter-individual variability of drug response. Five placebo-controlled trials of three structurally distinct selective inhibitors of COX-2 have revealed an increased hazard of myocardial infarction and stroke consistent with a mechanism-based class-specific cardiovascular hazard. Sustained inhibition of platelet TxA(2) by aspirin affords cardiovascular benefit, despite concomitant inhibition of PGI(2). Although there is no information from randomized placebo-controlled trials, traditional non-steroidal anti-inflammatory drugs, such as naproxen, dicofenac and ibuprofen, might differ in their effects of cardiovascular biology. PMID- 15780834 TI - Proteomic fingerprinting for the diagnosis of human African trypanosomiasis. AB - Papadopoulos et al. recently reported the discovery of a diagnostic serum proteomic signature for human African trypanosomiasis (HAT), using a combination of surface-enhanced laser desorption-ionization time-of-flight (SELDI-TOF) mass spectrometry and data-mining algorithms. This novel approach, coupled with biochemical characterization of the proteins that contribute to the signature, provides powerful new tools for the development of improved diagnostic tests, disease staging and identification of potential novel drug targets in HAT. PMID- 15780835 TI - Parasite role reversal: worms on trial. AB - Asthma has reached epidemic proportions globally. This has been attributed by many to improved hygiene. The frequent failure of conventional pharmaceuticals to manage the disease has led to the introduction of parasites as a potential alternative therapy for asthma and other immunological diseases. In this article, we briefly review the immunological rationale underpinning therapeutic parasitic infection, describe recently initiated trials, and highlight potential risks and benefits of introducing parasites into patient cohorts. PMID- 15780836 TI - Human balantidiasis in Iran: an unresolved enigma? PMID- 15780837 TI - Towards a new conceptual approach to "parasitoproteomics". AB - Many parasitologists are betting heavily on proteomic studies to explain biochemical host-parasite interactions and, thus, to contribute to disease control. However, many "parasitoproteomic" studies are performed with powerful techniques but without a conceptual approach to determine whether the host genomic responses during a parasite infection represent a nonspecific response that might be induced by any parasite or any other stress. In this article, a new conceptual approach, based on evolutionary concepts of immune responses of a host to a parasite, is suggested for parasitologists to study the host proteome reaction after parasite invasion. Also, this new conceptual approach can be used to study other host-parasite interactions such as behavioral manipulation. PMID- 15780838 TI - Does Trypanosoma cruzi calreticulin modulate the complement system and angiogenesis? AB - Calreticulin, a calcium-binding protein that is highly conserved in its multiple functions, is present in a wide spectrum of subcellular compartments in virtually every cell of higher organisms. In this article, we propose a dual role for parasite calreticulin, with emphasis on the Trypanosoma cruzi model. By modulating the vertebrate complement system, calreticulin might provide the parasite with an effective immune-escape mechanism. Alternatively, by inhibiting angiogenesis, the parasite molecule might protect the host from ongoing neoplasic aggressions. Many questions are still unanswered, particularly those regarding the consequences that these interactions could have in vivo for both the parasite and the host. PMID- 15780839 TI - IgE: a question of protective immunity in Trichinella spiralis infection. AB - The best-known function of immunoglobulin E (IgE) is the initiation of allergic reaction. However, it does not make sense that IgE exists only for allergy, which is harmful for the body and does not have a beneficial role. In this article, we focus on IgE in Trichinella spiralis infection in rodents and humans, and discuss the possible roles of IgE in protection and pathogenesis. PMID- 15780840 TI - Vaccination against canine babesiosis. AB - It has been known for several decades that the soluble parasite antigen (SPA) of several Babesia species can be used as a vaccine against the clinical manifestations of babesiosis. Originally observed in the plasma of infected animals, SPA can also be recovered from the supernatants of in vitro cultures of these parasites. Variable success has been reported for vaccines against the bovine and canine Babesia parasites, which seems to be related to antigenic diversity within Babesia species. In this article, an overview is presented of the development of such vaccines for dogs, and additional research that has led to improvement of an SPA-based vaccine against Babesia canis in dogs. PMID- 15780841 TI - Energy generation in insect stages of Trypanosoma brucei: metabolism in flux. AB - The generation of energy in African trypanosomes is a subject of undoubted importance. In bloodstream-form organisms, substrate-level phosphorylation of glucose is sufficient to provide the energy needs of the parasite. The situation in procyclic-form trypanosomes is more complex. For many years, it was accepted that glucose metabolism followed a conventional scheme involving glycolysis, the tricarboxylic acid cycle and ATP-producing oxidative phosphorylation linked to the electron-transport chain. However, progress in sequencing the Trypanosoma brucei genome and the development of gene-knockout and RNA interference technology has provided novel insight. Coupling these new technologies with classical approaches, including NMR and mass spectrometry to analyse glycolytic intermediates and end products, has yielded several surprises. In this article, we summarize how these recent data have helped to change the view of metabolism in procyclic-form T. brucei. PMID- 15780842 TI - Melanization immune responses in mosquito vectors. AB - The production and deposition of melanin pigments on invading pathogens and parasites represents a unique, innate immune response in the phylum Arthropoda. This immune response has started to receive considerable attention because of the potential to exploit this mechanism to control mosquito-borne diseases. In this article, we summarize knowledge about this complex biochemistry, the use of melanin biosynthesis in diverse physiological processes and the gaps in knowledge that must be addressed if this immune process is to be manipulated in genetic based control strategies. PMID- 15780843 TI - Antidepressant-like effects of cranial stimulation within a low-energy magnetic field in rats. AB - BACKGROUND: Evidence suggests that a novel type of magnetic resonance imaging (MRI) scan called echo planar magnetic resonance spectroscopic imaging (EP-MRSI) has mood-elevating actions in humans during the depressive phases of bipolar disorder. We examined whether a low-energy component of EP-MRSI (low-field magnetic stimulation [LFMS]) has antidepressant-like, locomotor-stimulating, or amnestic effects in rats. METHODS: We examined the effects of LFMS on immobility in the forced swim test (FST) and activity within an open field in separate groups of rats. After exposure to forced swimming, rats received LFMS (three 20 min sessions at 1.5 G/cm and .75 V/m) before behavioral testing. We also examined the effects of LFMS on fear conditioning (FC), a learning paradigm that also involves exposure to stressful conditions. RESULTS: Low-field magnetic stimulation reduced immobility in the FST, an antidepressant-like effect qualitatively similar to that of standard antidepressants. Low-field magnetic stimulation did not alter locomotor activity or FC. CONCLUSIONS: Low-field magnetic stimulation has antidepressant-like effects in rats that seem unrelated to locomotor-activating or amnestic effects. These findings raise the possibility that electromagnetic fields can affect the brain biology and might have physiologic consequences that offer novel approaches to therapy for psychiatric disorders. These same consequences might render MRI-based scans more invasive than previously appreciated. PMID- 15780844 TI - D-serine efficacy as add-on pharmacotherapy to risperidone and olanzapine for treatment-refractory schizophrenia. AB - BACKGROUND: D-serine, a selective full agonist at the glycine site of N-methyl-D aspartate glutamate receptor, might presently be the compound of choice for counteracting the hypothesized dysfunction of this receptor class in schizophrenia. Studies performed with Taiwanese patients indicate that D-serine significantly improves schizophrenia symptoms when used as adjuvant to conventional neuroleptics but not to clozapine. We assessed the efficacy and safety of D-serine adjuvant treatment for Occidental schizophrenia patients treated with newer atypical antipsychotics. METHODS: Thirty-nine risperidone- or olanzapine-treated schizophrenia patients participated in a double-blind, placebo controlled, 6-week crossover trial with 30 mg/kg/day D-serine added to their antipsychotic medication. Measures of clinical efficacy and side effects were determined biweekly throughout the study. Clinical laboratory parameters and amino acid serum levels were monitored. RESULTS: D-serine administration induced increased serine serum levels (p < .001) and resulted in significant (p < .001) improvements in negative, positive, cognitive, and depression symptoms, as measured by the Positive and Negative Syndrome Scale. For approximately one third of the sample, D-serine treatment resulted in significant (>20%) reductions in Brief Psychiatric Rating Scale total scores. D-serine was well tolerated, and no detrimental changes in clinical laboratory parameters were noted. CONCLUSIONS: These findings 1) indicate that risperidone and olanzapine efficacy might be augmented with D-serine adjuvant treatment; 2) confirm D-serine efficacy against main schizophrenia symptom domains; and 3) warrant the assessment of D-serine antipsychotic monotherapy for this illness. PMID- 15780845 TI - A putative animal model of the "prodromal" state of schizophrenia. AB - BACKGROUND: There is growing interest in detecting and treating schizophrenia during the "prodrome," before the symptoms are fully manifested. The objective of this study was to develop a putative model of the prodrome and study the effects of medications on it. METHODS: Rats were treated with different regimens of amphetamine to produce full sensitization (full syndrome) and partial sensitization (to model the prodromal state) and were then treated with typical and atypical antipsychotics and a D1 antagonist to mimic early intervention. After several weeks of withdrawal, locomotor activity in response to amphetamine and behavioral deficits (prepulse inhibition [PPI] and latent inhibition [LI]) were examined. RESULTS: Animals that received the full sensitization showed significant increase in locomotor activity and a disruption in both PPI and LI. Animals treated with a partial regimen showed only a muted phenotype. The animals that received "early intervention" did not show progression from the prodromal to the full-blown phenotype. CONCLUSIONS: The partial regimen of amphetamine injections provided a modified phenotype that could be regarded as a representative of the "prodromal" state. Early intervention, instituted once the prodromal state was already developed, prevented further progression into the full phenotype analogous to schizophrenia. PMID- 15780846 TI - Delta-9-tetrahydrocannabinol effects in schizophrenia: implications for cognition, psychosis, and addiction. AB - BACKGROUND: Recent advances in the neurobiology of cannabinoids have renewed interest in the association between cannabis and psychotic disorders. METHODS: In a 3-day, double-blind, randomized, placebo-controlled study, the behavioral, cognitive, motor, and endocrine effects of 0 mg, 2.5 mg, and 5 mg intravenous Delta-9-tetrahydrocannabinol (Delta-9-THC) were characterized in 13 stable, antipsychotic-treated schizophrenia patients. These data were compared with effects in healthy subjects reported elsewhere. RESULTS: Delta-9 tetrahydrocannabinol transiently increased 1) learning and recall deficits; 2) positive, negative, and general schizophrenia symptoms; 3) perceptual alterations; 4) akathisia, rigidity, and dyskinesia; 5) deficits in vigilance; and 6) plasma prolactin and cortisol. Schizophrenia patients were more vulnerable to Delta-9-THC effects on recall relative to control subjects. There were no serious short- or long-term adverse events associated with study participation. CONCLUSIONS: Delta-9-tetrahydrocannabinol is associated with transient exacerbation in core psychotic and cognitive deficits in schizophrenia. These data do not provide a reason to explain why schizophrenia patients use or misuse cannabis. Furthermore, Delta-9-THC might differentially affect schizophrenia patients relative to control subjects. Finally, the enhanced sensitivity to the cognitive effects of Delta-9-THC warrants further study into whether brain cannabinoid receptor dysfunction contributes to the pathophysiology of the cognitive deficits associated with schizophrenia. PMID- 15780847 TI - The neuropsychological basis of competence to consent in first-episode schizophrenia: a pilot metacognitive study. AB - BACKGROUND: This study was designed to explore the neuropsychological basis of competence to consent to treatment in first-episode schizophrenia by evaluating its differential and joint links with cognitive versus metacognitive performance. METHODS: Twenty-one first-episode patients were assessed with the MacArthur Competence Assessment Tool for Treatment (MacCAT-T) and a metacognitive version of the Wisconsin Card Sorting Test (WCST). In addition to the standard administration of the WCST, subjects were also asked to rate their level of confidence in the correctness of each sort (prior to getting the feedback) and to choose whether they wanted each sort to be "counted" toward their overall performance score on the test. Each "ventured" sort received a bonus of 10 cents if correct but an equal penalty if wrong. RESULTS: Compromised capacity to consent was more strongly related to deficits at the metacognitive level than to cognitive deficits per se. Moreover, prediction of competence to consent significantly improved when adding the new, free-choice metacognitive measures to the conventional WCST measures but not the other way around. CONCLUSIONS: These preliminary results suggest that metacognition plays a fundamental role in capacity to consent, which might be at least equally important for decision making competence as cognitive deficits per se. PMID- 15780848 TI - Clozapine and the mitogen-activated protein kinase signal transduction pathway: implications for antipsychotic actions. AB - BACKGROUND: Mitogen-activated protein kinase (MAPK) signaling pathways respond to dopaminergic and serotonergic agents and mediate short- and long-term effects of intracellular signaling in neurons. Here we show that the antipsychotic agent, clozapine, selectively activates the MEK/ERK MAPK pathway, and inhibition of this pathway reverses clozapine's actions in the conditioned avoidance response (CAR) paradigm, a rodent behavioral assay of antipsychotic activity. METHODS: Phosphorylation patterns of MAPK pathway enzymes were determined by quantitative immunoblot analysis and immunohistochemistry of rat prefrontal cortex. Kinase inhibitors were used to assess the role of MAPK signaling pathways in mediating clozapine-induced suppression of CAR. RESULTS: Clozapine administration selectively increased phosphorylation of MEK1/2 but had no effect on p38 or JNK phosphorylation. Pretreatment with the 5-HT2A agonist (+/-)-2,5-dimethoxy-4 iodoamphetamine hydrochloride blocked the clozapine-induced increase in MEK1/2 phosphorylation. Immunohistochemistry revealed that clozapine treatment elevated the number of cells in the prefrontal cortex positive for phosphoERK, the downstream substrate of MEK1/2. Prior administration of MEK1/2 inhibitors U0126 or Sl327, or ERK inhibitor 5-iodotubercidin, reversed suppression of CAR induced by clozapine, whereas administration of vehicle, JNK or p38 inhibitors (L-JNK-1 and SB203580, respectively) had no effect. Inhibition of kinases upstream to MEK1/2 (PI-3K, PKC, and CaMKII) by administration of LY294002, bisindolylmaleimide, or KN-62, respectively, also reversed clozapine-induced suppression of CAR. CONCLUSIONS: These data support the hypothesis that the MEK/ERK signal transduction cascade participates in clozapine's antipsychotic actions. PMID- 15780849 TI - Contributions of amygdala and striatal activity in emotion regulation. AB - BACKGROUND: Emotional information can facilitate or interfere with cognitive processes. In this study, we examined the influence of emotional information in biasing performance and the biological basis underlying this influence. METHODS: Ten human subjects (five female) were scanned with functional magnetic resonance imaging while performing an emotional go/nogo task. RESULTS: Subjects were slower to approach fearful target expressions and had more difficulty avoiding happy nontarget expressions. The amygdala was recruited most for negative emotional context, and activity in this region was positively correlated with response time when detecting negative expressions. Increased signal in the right caudate nucleus was observed when avoiding nontargets and was negatively correlated with the number of false alarms subjects made. CONCLUSIONS: Emotional context can alter behavioral and biological responses when approaching or avoiding a stimulus. We showed that recruitment of the amygdala, a region implicated in evaluating emotional significance, was associated with longer response latencies when approaching negative information, whereas recruitment of the caudate nucleus, a structure previously implicated in reward and impulse control, was most active when avoiding positive information. Our findings have significant implications for exaggerated and inhibited emotional responses that are characteristic of a number of psychiatric disorders. PMID- 15780850 TI - Hippocampal and ventricular volumes in psychotic and nonpsychotic bipolar patients compared with schizophrenia patients and community control subjects: a pilot study. AB - BACKGROUND: Previous reports of ventricular and hippocampal volumes in patients with bipolar disorder (BP) have been inconsistent in their findings. One possibility is that volumetric abnormalities are determined by disease subtype. Prior evidence suggests that psychotic (PBP) and nonpsychotic (NPBP) forms of BP are two subtypes that might differ in pathophysiology. METHODS: We investigated ventricular and hippocampal volumes in 38 adults with clearly defined PBP (n = 23) and NPBP subtypes, compared with 33 persons with schizophrenia (SZ) and 44 healthy community control subjects (HC). Ventricular and hippocampal volumes were reliably measured on high-resolution anatomic magnetic resonance imaging scans. We used a multivariate analysis of covariance to compare volumes across groups, covarying for total brain volume. Potential effects of BP illness features were explored, contrasting PBP and NPBP. RESULTS: For ventricular but not hippocampal regions, we found significant volume difference in PBP but not NPBP compared with HC (p < .005). We also observed nonsignificantly smaller left hippocampal volumes in PBP versus HC. Schizophrenic subjects had significantly larger ventricular and smaller left hippocampal volumes than HC. CONCLUSIONS: These results suggest that PBP but not NPBP is associated with increased ventricle volumes and a trend toward smaller left hippocampal volumes, as observed in SZ. PMID- 15780851 TI - The slow and long-lasting blockade of dopamine transporters in human brain induced by the new antidepressant drug radafaxine predict poor reinforcing effects. AB - BACKGROUND: (2S,3S)-2-(3-Chlorophenyl)-3,5,5,-trimethyl-2-morpholinol hydrochloride (radafaxine) is a new antidepressant that blocks dopamine transporters (DAT). A concern with drugs that block (DAT) is their potential reinforcing effects and abuse liability. Using positron emission tomography (PET) we have shown that for DAT-blocking drugs to produce reinforcing effects they must induce >50% DAT blockade and the blockade has to be fast (within 15 minutes). This study measures the potency and kinetics for DAT blockade by radafaxine in human brain. METHODS: PET and [11C]cocaine were used to estimate DAT blockade at 1, 4, 8, and 24 hours after radafaxine (40 mg p.o.) in 8 controls. Plasma pharmacokinetics and behavioral and cardiovascular effects were measured in parallel. RESULTS: DAT blockade by radafaxine was slow, and at 1 hour, it was 11%. Peak blockade occurred at about 4 hours and was 22%. Blockade was long lasting: at 8 hours 17%, and at 24 hours 15%. Peak plasma concentration occurred about 4 to 8 hours. No behavioral or cardiovascular effects were observed. CONCLUSIONS: The relatively low potency of radafaxine in blocking DAT and its slow blockade suggests that it is unlikely to have reinforcing effects. This is consistent with preclinical studies showing no self-administration. This is the first utilization of PET to predict abuse liability of a new antidepressant in humans based on DAT occupancy and pharmacokinetics. PMID- 15780852 TI - Genetic influences on the stability of attention-deficit/hyperactivity disorder symptoms from early to middle childhood. AB - BACKGROUND: The high heritability of the core symptoms of attention deficit/hyperactivity disorder (ADHD) has been repeatedly demonstrated, but few studies to date have investigated the extent to which the same genetic influences operate across development or new genes emerge at different developmental periods. METHODS: We report data from a large, population-based study of approximately 4,000 twin pairs, who have been followed up from early to middle childhood. RESULTS: Parents' ratings of ADHD symptoms showed moderate stability across the ages, which was mainly due to shared genetic influences. There was also evidence of additional genetic influences, which were not shared with those acting earlier on, emerging at later age periods. The contribution of environmental influences to the stability of the ADHD symptoms over time was small. Parents' ratings on the Conners' DSM-IV ADHD subscale at the last assessment point, at an average age of 8 years, did not show the rater contrast effects that were observed in the parents' ratings at earlier ages with briefer measures. Similar estimates of genetic and environmental influences were obtained for girls and boys. CONCLUSIONS: We discuss the implications of the findings for molecular genetic studies on ADHD symptomatology. PMID- 15780853 TI - Intergenerational transmission of subthreshold autistic traits in the general population. AB - BACKGROUND: Autistic disorder (AD) is a disabling oligogenic condition characterized by severe social impairment. Subthreshold autistic social impairments are known to aggregate in the family members of autistic probands; therefore, we conducted this study to examine the intergenerational transmission of such traits in the general population. METHODS: The Social Responsiveness Scale (SRS), a quantitative measure of autistic traits, was completed on 285 pairs of twins (by maternal report) and on their parents (by spouse report). RESULTS: Correlation for social impairment or competence between parents and their children and between spouses was on the order of .4. In families in which both parents scored in the upper quartile for social impairment on the SRS, mean SRS score of offspring was significantly elevated (effect size 1.5). Estimated assortative mating explained approximately 30% of the variation in parent SRS scores. CONCLUSIONS: Children from families in which both parents manifest subthreshold autistic traits exhibit a substantial shift in the distribution of their scores for impairment in reciprocal social behavior, toward the pathological end. As has been previously demonstrated in children, heritable subthreshold autistic impairments are measurable in adults and appear continuously distributed in the general population. PMID- 15780854 TI - Hyperactivity in adolescents born very preterm is associated with decreased caudate volume. AB - BACKGROUND: Several studies have associated preterm birth with impaired behavioral functioning and attention problems. In addition, preterm individuals have an increased risk of brain injury in the neonatal period. Such early lesions have the potential to disrupt subsequent neurodevelopment. This study explored behavioral functioning, particularly externalizing behavior, in a group of adolescents who were born very preterm and its relationship with volume of the caudate, a brain region particularly vulnerable to damage in the preterm neonate. METHODS: We studied 72 adolescents born before 33 weeks and 50 age- and gender matched full-term control subjects. Behavioral assessment included the Rutter Behavioural Scale and a social adjustment scale. Bilateral caudate volumes were quantified by stereologic methods. RESULTS: Preterm adolescents scored significantly higher than control subjects on the Rutter hyperactivity score, and boys scored higher than girls. In preterm boys only, left caudate volume was negatively correlated with hyperactivity score (r = -.43, p = .018) and social adjustment score in childhood (r = -.40, p = .028). CONCLUSIONS: Our data suggest that boys born very preterm are more likely to experience nonclinical behavioral problems in adolescence compared with full-term control subjects. Our results indicate that behavioral problems in this group might be associated with reductions in volume of the left caudate nucleus. PMID- 15780855 TI - Increased serum levels of interleukin-12 and tumor necrosis factor-alpha in Tourette's syndrome. AB - BACKGROUND: The hypothesis that common infections can modulate the onset and course of tic disorders and early-onset obsessive-compulsive disorder (OCD) in pediatric populations is longstanding. To date, most investigations have focused on the hypothesis of molecular mimicry and humoral immune responses. This study was carried out to investigate whether cytokines associated with the innate immune response or T cell activation were altered under baseline conditions and during periods of symptom exacerbation. METHODS: Forty-six patients with Tourette's syndrome and/or early-onset OCD, aged 7-17 years, and 31 age-matched control subjects participated in a prospective longitudinal study. Ratings of clinical severity and serum were collected at regular intervals, and serum concentrations of 10 cytokines were measured repeatedly. RESULTS: Interleukin-12 and tumor necrosis factor alpha concentrations at baseline were elevated in patients compared with control subjects. Both of these markers were further increased during periods of symptom exacerbation. CONCLUSIONS: These findings suggest that symptom exacerbations are associated with an inflammatory process propagated by systemic and local cytokine synthesis that might involve the central nervous system. We conclude that, in the future, longitudinal studies of children with neuropsychiatric disorders should examine the involvement of innate and T cell immunity. PMID- 15780856 TI - Plasma tryptophan during weight restoration in patients with anorexia nervosa. AB - BACKGROUND: Anorexia nervosa (AN) is a mental disorder characterized by low weight and concerns about body shape and weight. Disturbance in serotonin function has been described as central to the psychobiology of this disorder. Plasma tryptophan (TRP), the essential amino acid needed for serotonin production, is known to be low following acute caloric restriction but has not been measured during the course of refeeding. METHODS: Plasma TRP and other large neutral amino acids (LNAA) levels were measured in 26 female patients with AN and 15 control subjects. Patient levels were measured at admission for inpatient treatment, after 1 week of treatment, and upon weight restoration to weight > or =90% ideal body weight (IBW). For 17 patients, an additional assessment was made when weight reached 80% IBW. Plasma levels were obtained on one occasion from healthy control subjects. RESULTS: Plasma TRP and TRP/LNAA ratio increased significantly during refeeding process. Plasma TRP in patients was 46.88 nmol/mL (SD = 19.59) on admission and 55.54 nmol/L/mL (SD = 8.1) at normal weight, p < .05. The ratio of TRP to LNAA was .11 (SD = .03) on admission and .13 (SD = .02) at normal weight, p < .05. Plasma TRP is significantly lower in low-weight patients than in healthy control subjects (TRP = 53.73 [SD = 8.21]), but there was no significant difference between control subjects and normal-weight patients. CONCLUSIONS: Plasma TRP normalizes during the course of refeeding, supporting the hypothesis that serotonin function is disturbed in patients with anorexia nervosa. PMID- 15780858 TI - Defining a cognitive function decrement in schizophrenia. AB - BACKGROUND: Although cognitive impairment is described as a core component of the characterization of schizophrenia, a sizable percentage of patients are classified as unimpaired by traditional definitions of impairment. The purpose of this study was to determine the percentage of patients with schizophrenia meeting criteria for a "cognitive function decrement" defined as a current level of cognitive function that falls below the level predicted by premorbid estimates. METHODS: Linear regression analyses were performed on a healthy control population to determine a predicted composite cognitive score based on maternal education, paternal education, and reading score as indicators of premorbid intellectual function. The percentages of patients with current cognitive function above and below predicted values were calculated. RESULTS: When the Wide Range Achievement Test-3 (WRAT-3) score and maternal education are both used to predict current cognitive performance, as expected, about half (42%) of control subjects fall below expectations. However, 98.1 % of patients fall below expectations. CONCLUSIONS: When cognitive function decrement is defined as a failure to reach the expected level of cognitive functioning, almost all patients with schizophrenia meet this definition. PMID- 15780859 TI - A spectral framework for sperm shape characterization. AB - A novel methodology for characterization of animal sperm shape involving the use of a spectral approach to multiscale curvature estimation is proposed. By using the derivative property of the Fourier transform, allied to Gaussian smoothing, accurate estimates of the curvature along the sperm contour can be obtained in such a way that the curvature peaks corresponding to the sperm head vertices can be effectively identified. The measurements derived from such a processing, namely the width of the basal region of the head, the centralization of tail implantation, and the multiscale bending energy, provide valuable resources for fertility and phylogenetic studies. PMID- 15780860 TI - An algorithmic approach to understand trace elemental homeostasis in serum samples of Parkinson disease. AB - A classical problem in neurological disorders is to understand the progression of disorder and define the trace elements (metals) which play a role in deviating a sample from normal to an abnormal state, which implies the need to create a reference knowledge base (KB) employing the control samples drawn from normal/healthy set in the context of the said neurological disorder, and in sequel to analytically understand the deviations in the cases of disorders/abnormalities/unhealthy samples. Hence building up a computational model involves mining the healthy control samples to create a suitable reference KB and designing an algorithm for estimating the deviation in case of unhealthy samples. This leads to realizing an algorithmic cognition-recognition model, where the cognition stage establishes a reference model of a normal/healthy class and the recognition stage involves discriminating whether a given test sample belongs to a normal class or not. Further if the sample belongs to a specified reference base (normal) then the requirement is to understand how strong the affiliation is, and if otherwise (abnormal) how far away the sample is from the said reference base. In this paper, an exploratory data analysis based model is proposed to carry out such estimation analysis by designing distribution and parametric models for the reference base. Further, the knowledge of the reference base in case of the distribution model is expressed in terms of zones with each zone carrying a weightage factor. Different distance measures are utilized for the subsequent affiliation analysis (City block with distribution model and Doyle's with Parametric model). Results of an experimental study based on the database of trace elemental analysis in human serum samples from control and Parkinson's neurological disorder are presented to corroborate the performance of the computational algorithm. PMID- 15780857 TI - Behavioral heterogeneity in an animal model of neuropsychiatric lupus. AB - BACKGROUND: Various psychiatric manifestations of unknown etiology are common in systemic autoimmune disease lupus erythematosus (SLE). Profound heterogeneity at clinical and neuropathological levels suggests distinct subpopulations of SLE patients and multiple mechanisms in the pathogenesis of aberrant behavior. Using inbred mice prone to SLE-like condition, we presently examine whether subpopulations of diseased mice can be identified on the basis of their behavioral performance. METHODS: Hierarchical cluster analysis was used to classify 105 MRL-lpr males into clusters. Multivariate analysis of variance (MANOVA) and discriminant function analysis were used to detect overall differences and identify discriminative variables. RESULTS: Cluster 1 was characterized by blunted responsiveness to palatable stimulation, as well as increased spleen mass and serum levels of interleukin-1. Cluster 2 comprised of animals with reduced ambulation speed and enlarged spleen. Mice from cluster 3 showed profound dilatation of brain ventricles, reduced brain mass, impaired nutrition and performance in task reflective of emotional reactivity. CONCLUSIONS: Present results suggest that systemic autoimmunity compromises brain function via non-Mendelian mechanisms. Although neuroactive cytokines may impair reward systems, brain atrophy seems to underlie deficits in ingestive behavior and emotional reactivity. This study supports the hypothesis that multiple neuroimmunological pathways are involved in the etiology of aberrant behavior during SLE-like disease. PMID- 15780861 TI - A multi-channel adaptive nonlinear filtering structure realizing some properties of the hearing system. AB - An adaptive nonlinear signal-filtering model of the cochlea is proposed based on the functional properties of the inner ear. The model consists of the cochlear filtering segments taking into account the longitudinal, transverse and radial pressure wave propagation. On the basis of an analytical description of different parts of the model and the results of computer modeling, the biological significance of the nonlinearity of signal transduction processes in the outer hair cells, their role in signal compression and adaptation, the efferent control over the characteristics of the filtering structures (frequency selectivity and sensitivity) are explained. PMID- 15780862 TI - Computer phantom study of brain PET glucose metabolism imaging using a rotating SPECT/PET camera. AB - Positron emission tomography (PET) with [18F] fluoro-deoxy-glucose (FDG) provides information about glucose metabolism and is used to measure tissue glucose kinetics in the brain. The recent interest in hybrid SPECT/PET systems emerged as a practical approach to reduce the high cost of purchasing a dedicated ring detector PET system. We have implemented interpolation methods for processing the projection data that could potentially reduce artifacts when reconstructing a dynamic imaging sequence in a PET study from a dual-head rotating SPECT/PET system. The computer simulations predict that parameter estimates from the dedicated PET system will be superior to results using the rotating camera system. However, the rotating camera system using projection interpolation may approach the accuracy of the dedicated PET system if the data noise is below 20%. PMID- 15780863 TI - Improving medical diagnostic accuracy of ultrasound Doppler signals by combining neural network models. AB - There are a number of different quantitative models that can be used in a medical diagnostic decision support system including parametric methods (linear discriminant analysis or logistic regression), nonparametric models (k nearest neighbor or kernel density) and several neural network models. The complexity of the diagnostic task is thought to be one of the prime determinants of model selection. Unfortunately, there is no theory available to guide model selection. This paper illustrates the use of combined neural network models to guide model selection for diagnosis of ophthalmic and internal carotid arterial disorders. The ophthalmic and internal carotid arterial Doppler signals were decomposed into time-frequency representations using discrete wavelet transform and statistical features were calculated to depict their distribution. The first-level networks were implemented for the diagnosis of ophthalmic and internal carotid arterial disorders using the statistical features as inputs. To improve diagnostic accuracy, the second-level networks were trained using the outputs of the first level networks as input data. The combined neural network models achieved accuracy rates which were higher than that of the stand-alone neural network models. PMID- 15780864 TI - Cytidine deamination and resistance to retroviral infection: towards a structural understanding of the APOBEC proteins. AB - The human apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G (APOBEC3G, or hA3G) protein, provides cells with an intracellular antiretroviral activity that is associated with the hypermutation of viral DNA through cytidine deamination. Indeed, hA3G belongs to a family of vertebrate proteins that contain one or two copies of a signature sequence motif unique to cytidine deaminases (CTDAs). We have constructed secondary structure models of the APOBEC proteins through a combination of structure prediction and subsequent alignment with nucleotide CTDAs whose structures have been solved to high resolution. Secondary structure elements common to all CTDAs are predicted, in addition to structural features that are apparently unique to the APOBEC family of proteins. Most notably, a putative looped-out helix abuts an amino acid that modulates the susceptibility of A3G proteins to the antagonistic action of the human and simian immunodeficiency virus (HIV and SIV) Vif proteins. Using the structure models as a guide, we reflect on mutagenesis studies of the APOBEC1 (A1), hA3G and activation induced deaminase (AID) proteins, with emphasis on the determinants of cytidine deamination and antiviral activities. PMID- 15780865 TI - Virus evolution during a severe dengue epidemic in Cuba, 1997. AB - Full-length genomic sequences from six DENV-2 isolates sampled at different times during a dengue outbreak that occurred in Cuba in 1997 were determined. Phylogenetic analysis indicated that these isolates fall into the "American/Asian" genotype. Genome analysis revealed strong conservation of the structural proteins and the non-coding regions (5' NCR and 3' NCR). Nucleotide substitutions were observed in non-structural genes and most notably in the NS5 gene. There was a clear pattern of virus evolution during the epidemic; the earliest isolates sampled differed from those sampled later by amino acid replacements in the NS1 and NS5 proteins, although there was no evidence that these represented escape mutants. Further studies are therefore required to define the functional role of amino acid replacements observed and their possible relation to disease severity. PMID- 15780866 TI - Neutralizing antibody and protective immunity to SARS coronavirus infection of mice induced by a soluble recombinant polypeptide containing an N-terminal segment of the spike glycoprotein. AB - A secreted, glycosylated polypeptide containing amino acids 14 to 762 of the SARS coronavirus (SARS-CoV) spike protein and a polyhistidine tag was expressed in recombinant baculovirus-infected insect cells. Mice received the affinity purified protein with either a saponin (QS21) or a Ribi (MPL + TDM) adjuvant subcutaneously and were challenged intranasally with SARS-CoV. Both regimens induced binding and neutralizing antibodies and protection against SARS-CoV intranasal infection. However, the best results were obtained with QS21 and protein, which provided the highest antibody as well as complete protection of the upper and lower respiratory tract. PMID- 15780867 TI - Characterization and regulation of essential murine cytomegalovirus genes m142 and m143. AB - US22 gene family members m142 and m143 are essential for replication of murine cytomegalovirus (MCMV). Their transcripts are produced with immediate-early kinetics, but little else is known about these viral genes. Unlike their transcripts, the m142 and m143 gene products (pm142, pm143) were not expressed until early times post-infection, with levels increasing over the course of infection. Both pm142 and pm143 were predominantly cytoplasmic, but cellular fractionation studies confirmed that the proteins were present in the nucleus as well. In addition, pm142 was detected within the virion. Both the m142 and m143 promoters were strongly upregulated by viral infection or by MCMV IE1. However, UV-inactivated virus and IE3 upregulated only the m142 promoter. When tested for transcriptional transactivating activity, neither m142 nor m143 demonstrated significant activity, either alone or in combination with the major immediate early gene products. This failure to transactivate, along with their essential nature, makes m142 and m143 unique among the immediate-early genes of the US22 gene family. PMID- 15780868 TI - Peripheral nerve-derived HIV-1 is predominantly CCR5-dependent and causes neuronal degeneration and neuroinflammation. AB - HIV-related peripheral neuropathy is a major neurological complication of HIV infection, although little is known about its pathogenesis. We amplified HIV-1 C2V3 envelope sequences from peroneal nerves obtained from HIV/AIDS patients. Sequence analysis and infectious recombinant viruses containing peripheral nerve derived C2V3 sequences indicated a predominance of CCR5-dependent and macrophage tropic HIV-1, although dual tropic viruses using both CCR5 and CXCR4 were identified. The neuropathogenic effects of recombinant HIV-1 clones were investigated using a novel dorsal root ganglion culture system that was comprised of sensory neurons, macrophages and Schwann cells from transgenic rats expressing human CD4 and CCR5 on monocytoid cells. Despite restricted viral replication, HIV 1 infection caused a reduction in the percentage of neurons with neuritic processes together with significant neurite retraction, which was accompanied by induction of IL-1beta and TNF-alpha expression, depending on the individual virus. Our results suggest that HIV-1 infection of the peripheral nervous system causes axonal degeneration, possibly through the induction of pro-inflammatory cytokines. PMID- 15780869 TI - Interaction of the adenovirus major core protein precursor, pVII, with the viral DNA packaging machinery. AB - Adenovirus is one of the well-studied double-stranded DNA viruses. However, the mechanisms of its DNA packaging and virion assembly are still not fully understood. One of the unique features of adenovirus is that the unpackaged viral DNA is associated with core protein pVII. Packaging of viral DNA bound with proteins has not been reported from other viruses. To characterize how viral DNA bound with protein pVII is packaged, we performed experiments to see if protein pVII interacts with the known DNA packaging proteins or the packaging sequence. Our results demonstrated that protein pVII interacted with the viral IVa2 and L1 52/55 kDa proteins, which are the known viral DNA packaging proteins. Furthermore, our protein-DNA binding experiments demonstrated that the IVa2 protein mediates the specific interaction with the packaging sequence, whereas protein pVII and the L1 52/55 kDa protein bind to DNA non-specifically. Although the non-specific binding of protein pVII and the L1 52/55 kDa protein do not appear to affect the specific binding of the IVa2 protein to the packaging sequence, and the specific binding of the IVa2 protein does not appear to block the bindings of protein pVII and the L1 52/55 kDa protein to the packaging sequence, the possibility of a cooperative binding among the IVa2 protein, the L1 52/55 kDa protein and protein pVII on the packaging sequence needs to be further determined. In summary, the results indicate that the assembly of the DNA packaging initiation complex may be mediated by the specific interaction of the IVa2 protein with the packaging sequence and other viral proteins, such as protein pVII and the L1 52/55 kDa protein. PMID- 15780870 TI - The distribution of pol containing human endogenous retroviruses in non-human primates. AB - Few human endogenous retroviruses (HERVs) have been extensively studied in non human primates. Such investigations have demonstrated that several element classes are primate unique, contain members with important biological function, are conserved in specific primate lineages, and have in some cases expanded in copy number. We have examined multiple sub-families of all major groups of HERVs using a DNA microarray based on the reverse transcriptase (RT) domain of the viral polymerase gene (pol). The microarray was used to investigate the distribution of HERVs in non-human primates with particular focus on the differences between New World monkeys (NWMs) and other anthropoids. This is the first study examining most HERV families in multiple non-human primate DNAs using a uniform and sensitive method and suggests that major differences exist between primate groups. The results indicate that a major invasion and expansion of pol containing HERVs occurred after the platyrrhine (NWM) lineage separated from the catarrhines (Old World Monkeys and apes). PMID- 15780871 TI - Potent inhibition of HIV-1 entry by (s4dU)35. AB - We have previously reported the potent in vitro HIV-1 anti-reverse transcriptase activity of a 35-mer of 4-thio-deoxyuridylate [(s(4)dU)(35)]. In efforts to define its activity in a more physiological system, studies were carried out to determine the stage of viral infection that this compound mediates its anti-viral effect. Results of the studies reported herein show that (s(4)dU)(35) is nontoxic and is capable of inhibiting both single and multi-drug resistant HIV strains (IC(50): 0.8-25.4 microg/ml) in vitro. Besides its previously reported anti-RT activity, (s(4)dU)(35) mediated its antiviral action by preventing virus attachment (IC(50): 0.002-0.003 microg/ml), and was stable in vitro and slowly degraded by DNAses. Competition studies and fluorescence resonance energy transfer (FRET) experiments indicated that (s(4)dU)(35) preferentially binds to CD4 receptors, but not to CD48. Confocal laser scanning microscopy (CLSM) studies showed that (s(4)dU)(35) did not penetrate into the cells and colocalized with cell surface thioredoxin. Our studies identify (s(4)dU)(35) as a potential novel HIV entry inhibitor that may have utility as either a systemic antiretroviral or as a preventing agent for HIV transmission. PMID- 15780872 TI - Identification and characterization of the pseudorabies virus UL43 protein. AB - Among the least characterized herpesvirus membrane proteins are the homologs of UL43 of herpes simplex virus 1 (HSV-1). To identify and characterize the UL43 protein of pseudorabies virus (PrV), part of the open reading frame was expressed in Escherichia coli and used for immunization of a rabbit. The antiserum recognized in Western blots a 34-kDa protein in lysates of PrV infected cells and purified virions, demonstrating that the UL43 protein is a virion component. In indirect immunofluorescence analysis, the antiserum labeled vesicular structures in PrV infected cells which also contained glycoprotein B. To functionally analyze UL43, a deletion mutant was constructed lacking amino acids 23-332 of the 373aa protein. This mutant was only slightly impaired in replication as assayed by one-step growth kinetics, measurement of plaque sizes, and electron microscopy. Interestingly, the PrV UL43 protein was able to inhibit fusion induced by PrV glycoproteins in a transient expression-fusion assay to a similar extent as gM. Double mutant viruses lacking, in addition to UL43, the multiply membrane spanning glycoproteins K or M did not show a phenotype beyond that observed in the gK and gM single deletion mutants. PMID- 15780873 TI - Distinct roles of enhancer nuclear factor 1 (NF1) sites in plasmacytoma and osteopetrosis induction by Akv1-99 murine leukemia virus. AB - Murine leukemia viruses (MLVs) can be lymphomagenic and bone pathogenic. In this work, the possible roles of two distinct proviral enhancer nuclear factor 1 (NF1) binding sites in osteopetrosis and tumor induction by B-lymphomagenic Akv1-99 MLV were investigated. Akv1-99 and mutants either with NF1 site 1, NF1 site 2 or both sites disrupted induced tumors (plasma cell proliferations by histopathology) with remarkably similar incidence and mean latency in inbred NMRI mice. Clonal immunoglobulin gene rearrangement detection, by Southern analysis, confirmed approximately half of the tumors induced by each virus to be plasmacytomas while the remaining lacked detectable clonally rearranged Ig genes and were considered polyclonal; a demonstration that enhancer NF1 sites are dispensable for plasmacytoma induction by Akv1-99. In contrast, X-ray analysis revealed significant differences in osteopetrosis induction by the four viruses strongly indicating that NF1 site 2 is critical for viral bone pathogenicity, whereas NF1 site 1 is neutral or moderately inhibitory. In conclusion, enhancer NF1 sites are major determinants of osteopetrosis induction by Akv1-99 without significant influence on viral oncogenicity. PMID- 15780874 TI - The RNA of turnip yellow mosaic virus exhibits icosahedral order. AB - Difference electron density maps, based on structure factor amplitudes and experimental phases from crystals of wild-type turnip yellow mosaic virus and those of empty capsids prepared by freeze-thawing, show a large portion of the encapsidated RNA to have an icosahedral distribution. Four unique segments of base-paired, double-helical RNA, one to two turns in length, lie between 33-A and 101-A radius and are organized about either 2-fold or 5-fold icosahedral axes. In addition, single-stranded loops of RNA invade the pentameric and hexameric capsomeres where they contact the interior capsid surface. The remaining RNA, not seen in electron density maps, must serve as connecting links between these secondary structural elements and is likely icosahedrally disordered. The distribution of RNA observed crystallographically appears to be in agreement with models based on biochemical data and secondary structural analyses. PMID- 15780875 TI - Endoplasmic reticulum targeting sequence enhances HBV-specific cytotoxic T lymphocytes induced by a CTL epitope-based DNA vaccine. AB - CD8(+) T cells play a critical role in protective immunity against Hepatitis B Virus (HBV). Epitope-based DNA vaccines expressing HBV-dominant CTL epitopes can be used as candidate vaccines capable of inducing cytotoxic T Lymphocytes (CTL) responses. A plasmid DNA encoding a CTL epitope of HBV core antigen, HBc(18-27), was constructed. Intramuscular immunization of C57BL/6 mice with this DNA vaccine resulted in successful induction of HBV-specific CTL responses. In order to promote transportation of the peptide into endoplasmic reticulum (ER) to bind to MHC class I molecules for optimal class I antigen presentation, an ER targeting sequence (ERTS) was fused with the C(18-27) encoding gene. ERTS fusion significantly enhanced specific CD8(+) T cell responses in terms of CTL cytolysis as well as IFN-gamma secretion. This enhancement was correlated with promoted epitope presentation on target cell surface. We report here an enhanced immunogenicity of an epitope-based DNA vaccine using an ER targeting signal sequence, which has significant implications for future design of therapeutic HBV vaccine. PMID- 15780876 TI - The antimicrobial peptide dermaseptin S4 inhibits HIV-1 infectivity in vitro. AB - Most of HIV-1 infections are acquired through sexual contact. In the absence of a preventive vaccine, the development of topical microbicides that can block infection at the mucosal tissues is needed. Dermaseptin S4 (DS4) is an antimicrobial peptide derived from amphibian skin, which displays a broad spectrum of activity against bacteria, yeast, filamentous fungi, and herpes simplex virus type 1. We show here that DS4 inhibits cell-free and cell associated HIV-1 infection of P4-CCR5 indicator cells and human primary T lymphocytes. The peptide is effective against R5 and X4 primary isolates and laboratory-adapted strains of HIV-1. Its activity is directed against HIV-1 particles by disrupting the virion integrity. Increasing the number of DS4 positive charges reduced cytotoxicity without affecting the antiviral activity. The modified DS4 inhibited HIV-1 capture by dendritic cells and subsequent transmission to CD4(+) T cells, as well as HIV-1 binding on HEC-1 endometrial cells and transcytosis through a tight epithelial monolayer. PMID- 15780878 TI - Nucleocytoplasmic shuttling of the rabies virus P protein requires a nuclear localization signal and a CRM1-dependent nuclear export signal. AB - Rabies virus P protein is a co-factor of the viral RNA polymerase. It has been shown previously that P mRNA directs the synthesis of four N-terminally truncated P products P2, P3, P4, and P5 due to translational initiation by a leaky scanning mechanism at internal Met codons. Whereas P and P2 are located in the cytoplasm, P3, P4, and P5 are found in the nucleus. Here, we have analyzed the molecular basis of the subcellular localization of these proteins. Using deletion mutants fused to GFP protein, we show the presence of a nuclear localization signal (NLS) in the C-terminal part of P (172-297). This domain contains a short lysine-rich stretch ((211)KKYK(214)) located in close proximity with arginine 260 as revealed by the crystal structure of P. We demonstrate the critical role of lysine 214 and arginine 260 in NLS activity. In the presence of Leptomycin B, P is retained in the nucleus indicating that it contains a CRM1-dependent nuclear export signal (NES). The subcellular distribution of P deletion mutants indicates that the domain responsible for export is the amino-terminal part of the protein. The use of fusion proteins that have amino terminal fragments of P fused to beta galactosidase containing the NLS of SV40 T antigen allows us to identify a NES between residues 49 and 58. The localization of NLS and NES determines the cellular distribution of the P gene products. PMID- 15780877 TI - Receptor specificity of influenza viruses from birds and mammals: new data on involvement of the inner fragments of the carbohydrate chain. AB - We studied receptor-binding properties of influenza virus isolates from birds and mammals using polymeric conjugates of sialooligosaccharides terminated with common Neu5Ac alpha2-3Gal beta fragment but differing by the structure of the inner part of carbohydrate chain. Viruses isolated from distinct avian species differed by their recognition of the inner part of oligosaccharide receptor. Duck viruses displayed high affinity for receptors having beta1-3 rather than beta1-4 linkage between Neu5Ac alpha2-3Gal-disaccharide and penultimate N acetylhexosamine residue. Fucose and sulfate substituents at this residue had negative and low effect, respectively, on saccharide binding to duck viruses. By contrast, gull viruses preferentially bound to receptors bearing fucose at N acetylglucosamine residue, whereas chicken and mammalian viruses demonstrated increased affinity for oligosaccharides that harbored sulfo group at position 6 of (beta1-4)-linked GlcNAc. These data suggest that although all avian influenza viruses preferentially bind to Neu5Ac alpha2-3Gal-terminated receptors, the fine receptor specificity of the viruses varies depending on the avian species. Further studies are required to determine whether observed host-dependent differences in the receptor specificity of avian viruses can affect their ability to infect humans. PMID- 15780879 TI - HPV16, HPV18, and HIV infection may influence cervical cytokine intralesional levels. AB - Infection with oncogenic human papillomavirus (HPV) is considered to be the major risk to cervical cancer. This study analyzed the influence of HPV infection on cytokine intralesional levels in cervical lesion in the presence or not of HIV infection. Cervical biopsies from 42 women were studied. HPV detection and typing were performed using amplified DNA hybridized with sequence-specific primers, and cytokine intralesional levels were detected using ELISA. HPV16+ biopsies exhibited increased IFN-gamma and IL-10 when compared to HPV16- (P = 0.03 and 0.04, respectively). HPV18+ biopsies exhibited decreased TNF-alpha (P = 0.009) and IFN- gamma (P = 0.01) when compared to HPV18-. In accordance to HIV status, HIV-/HPV16+ patients exhibited increased IFN-gamma when compared to those presenting HIV-/HPV16- (P = 0.007). HIV-/HPV18+ patients presented decreased IFN gamma when compared to HIV-/HPV18- (P = 0.02). These results suggest that the presence of HPV16 infection may influence cervical lesion installation, and irrespective of HIV status, HPV18 infection may be more aggressive than HPV-16. PMID- 15780880 TI - Differential evolution of cell-associated virus in blood and genital tract of HIV infected females undergoing HAART. AB - PBMC and vaginal cell (VC) viruses were studied from 5 HIV-infected females for the presence of drug-resistance and non-drug resistance associated mutations. A 1318-bp fragment of polymerase gene was amplified from PBMC and VC proviral DNA. Four of the 5 PBMC viruses exhibited drug resistance-associated mutations in reverse transcriptase and protease genes, whereas only 2 VC viruses contained drug resistance-associated mutations. However, all 5 females showed non-drug resistance-associated mutations both in PBMC and VC virus suggesting continuous evolution of the virus in these compartments. The emergence of drug resistance was slower in PBMC and VC viruses than that observed in the cell-free plasma (P) and vaginal secretion (VS) viruses. Phylogenetic analysis revealed that VC virus was closer to PBMC virus than either cell-free viruses (P and VS) suggesting comparable evolution among cell-associated viruses. PMID- 15780881 TI - Spike protein assembly into the coronavirion: exploring the limits of its sequence requirements. AB - The coronavirus spike (S) protein, required for receptor binding and membrane fusion, is incorporated into the assembling virion by interactions with the viral membrane (M) protein. Earlier we showed that the ectodomain of the S protein is not involved in this process. Here we further defined the requirements of the S protein for virion incorporation. We show that the cytoplasmic domain, not the transmembrane domain, determines the association with the M protein and suffices to effect the incorporation into viral particles of chimeric spikes as well as of foreign viral glycoproteins. The essential sequence was mapped to the membrane proximal region of the cytoplasmic domain, which is also known to be of critical importance for the fusion function of the S protein. Consistently, only short C terminal truncations of the S protein were tolerated when introduced into the virus by targeted recombination. The important role of the about 38-residues cytoplasmic domain in the assembly of and membrane fusion by this approximately 1300 amino acids long protein is discussed. PMID- 15780882 TI - Human and rodent humoral immune responses to Andes virus structural proteins. AB - In the present work we identified B-cell epitopes recognized by sera of humans and rodents naturally infected with Andes virus, a hantavirus present in Chile and Argentina. Analysis of patient and rodent sera with overlapping peptides revealed 21 human and rodent epitopes on the three structural proteins. Whereas in the nucleoprotein the region comprising aa 248-260 was shown to be the key determinant of human sera, the major antigenic site of rodent antibody reactivity is located at aa 326-338. In G1, the main epitope recognized by human sera was mapped to aa 14-26, while rodent antibodies bound predominantly to aa 599-611. In contrast, humans and mice had strong responses to three regions in G2 (aa 691 703, aa 918-930, aa 955-967), of which the last two are associated with neutralization of Hantaan virus. This insight affords important information for the development of immunotherapies for the acute phase of hantavirus cardiopulmonary syndrome. PMID- 15780883 TI - Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp. AB - Here, we report for the first time the successful use of cycloheximide (CHX) as an inhibitor to block de novo viral protein synthesis during WSSV (white spot syndrome virus) infection. Sixty candidate IE (immediate-early) genes were identified using a global analysis microarray technique. RT-PCR showed that the genes corresponding to ORF126, ORF242 and ORF418 in the Taiwan isolate were consistently CHX-insensitive, and these genes were designated ie1, ie2 and ie3, respectively. The sequences for these IE genes also appear in the two other WSSV isolates that have been sequenced. Three corresponding ORFs were identified in the China WSSV isolate, but only an ORF corresponding to ie1 was predicted in the Thailand isolate. In a promoter activity assay in Sf9 insect cells using EGFP (enhanced green fluorescence protein) as a reporter, ie1 showed very strong promoter activity, producing higher EGFP signals than the insect Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV) ie2 promoter. PMID- 15780884 TI - The virion-associated Gag-Pol is decreased in chimeric Moloney murine leukemia viruses in which the readthrough region is replaced by the frameshift region of the human immunodeficiency virus type 1. AB - The human immunodeficiency virus type 1 (HIV-1) requires a programmed -1 translational frameshift event to synthesize the precursor of its enzymes, Gag Pol, when ribosomes from the infected cells translate the full-length viral messenger RNA. Translation of the same RNA according to conventional translational rules produces Gag, the precursor of the structural proteins of the virus. The efficiency of the frameshift controls the ratio of Gag-Pol to Gag, which is critical for viral infectivity. The Moloney murine leukemia virus (MoMuLV) uses a different strategy, the programmed readthrough of a stop codon, to synthesize Gag-Pol. In this study, we investigated whether different forms of the HIV-1 frameshift region can functionally replace the readthrough signal in MoMuLV. Chimeric proviral DNAs were obtained by inserting into the MoMuLV genome the HIV-1 frameshift region encompassing the slippery sequence where the frameshift occurs, followed by the frameshift stimulatory signal. The inserted signal was either a simple stem-loop, previously considered as the stimulatory signal, or a longer bulged helix, now shown to be the complete stimulatory signal, or a mutated version of the complete signal with a three-nucleotide deletion. Although the three chimeric viruses can propagate essentially as the wild-type virus in NIH 3T3 cells, single-round infectivity assays revealed that the infectivity of the chimeric virions is about three to fivefold lower than that of the wild-type virions, depending upon the nature of the frameshift signal. It was also observed that the Gag-Pol to Gag ratio was decreased about two to threefold in chimeric virions. Comparison of the readthrough efficiency of MoMuLV to the HIV-1 frameshift efficiency, by monitoring the expression of a luciferase reporter in cultured cells, revealed that the frameshift efficiencies were only 30-60% of the readthrough efficiency. Altogether, these observations indicate that replacement of the readthrough region of MoMuLV with the frameshift region of HIV-1 results in virions that are replication competent, although less infectious than wild-type MoMuLV. This type of chimera could provide an interesting tool for in vivo studies of novel drugs targeted against the HIV-1 frameshift event. PMID- 15780885 TI - Piecewise linear correction of ECG baseline wander: a curve simplification approach. AB - In this paper, we suggest a novel method for ECG baseline correction, exclusively based on pattern recognition tools, namely, dominant points (DPs). The DPs are computed by the Douglas-Peucker curve simplification algorithm. The so-computed DPs include peak and baseline points, the discrimination of which yields gradual piecewise linear estimation of the baseline wander (BaselineW) in two iterations. At each iteration, the current BaselineW is subtracted from the input signal according to the decomposition scheme: ECG approximately ECG(ZBLW) + BaselineW, where ECG(ZBLW) is the underlying baseline wander free ECG. The method targets many types of baseline deviations in a unified approach: baseline drift due to respiration, amplitude modulation due to perspiration and abrupt potential change due to electrode loose contact. We tested the developed method on a variety of ECG records including half synthesized records contaminated with different types of baseline deviations (simulated) noise, and on records from the MITBIH database presenting important baseline deviations, including normal and abnormal heart beats cases. The method showed good performance in computing a piecewise linear estimation of the baseline deviation and in extracting the ECG(ZBLW), which represents the clinically significant electrocardiogram information. PMID- 15780886 TI - TERMTrial--terminology-based documentation systems for cooperative clinical trials. AB - Within cooperative groups of multi-center clinical trials a standardized documentation is a prerequisite for communication and sharing of data. Standardizing documentation systems means standardizing the underlying terminology. The management and consistent application of terminology systems is a difficult and fault-prone task, which should be supported by appropriate software tools. Today, documentation systems for clinical trials are often implemented as so-called Remote-Data-Entry-Systems (RDE-systems). Although there are many commercial systems, which support the development of RDE-systems there is none offering a comprehensive terminological support. Therefore, we developed the software system TERMTrial which consists of a component for the definition and management of terminology systems for cooperative groups of clinical trials and two components for the terminology-based automatic generation of trial databases and terminology-based interactive design of electronic case report forms (eCRFs). TERMTrial combines the advantages of remote data entry with a comprehensive terminological control. PMID- 15780887 TI - Rapid application design of an electronic clinical skills portfolio for undergraduate medical students. AB - The aim was to find how to use information and communication technology to present the clinical skills content of an undergraduate medical curriculum. Rapid application design was used to develop the product, and technical action research was used to evaluate the development process. A clinician-educator, two medical students, two computing science masters students, two other project workers, and a hospital education informatics lead, formed a design team. A sample of stakeholders took part in requirements planning workshops and continued to advise the team throughout the project. A university hospital had many features that favoured fast, inexpensive, and successful system development: a clearly defined and readily accessible user group; location of the development process close to end-users; fast, informal communication; leadership by highly motivated and senior end-users; devolved authority and lack of any rigidly imposed management structure; cooperation of clinicians because the project drew on their clinical expertise to achieve scholastic goals; a culture of learning and involvement of highly motivated students. A detailed specification was developed through storyboarding, use case diagramming, and evolutionary prototyping. A very usable working product was developed within weeks. "SkillsBase" is a database web application using Microsoft Active Server Pages, served from a Microsoft Windows 2000 Server operating system running Internet Information Server 5.0. Graphing functionality is provided by the KavaChart applet. It presents the skills curriculum, provides a password-protected portfolio function, and offers training materials. The curriculum can be presented in several different ways to help students reflect on their objectives and progress towards achieving them. The reflective portfolio function is entirely private to each student user and allows them to document their progress in attaining skills, as judged by self, peer and tutor assessment, and examinations. Training materials include web links and materials developed locally using pedagogic principles developed by the SkillsBase team. Although the usability of SkillsBase has been proven, uptake of software that has arisen 'bottom-up' from within the curriculum has proved slow. We plan to incorporate the SkillsBase services into a more comprehensive virtual managed learning environment, anticipating that presenting the functionality in an environment that is routinely used by students and teachers will increase uptake and use. PMID- 15780888 TI - A robust method for alignment of histological images. AB - Determination of reference points is a precondition for reconstruction of serial sections. In the case of comprehensive reconstruction work, manual extraction of the markers may be very time-consuming and may even make such reconstruction impossible. The procedure presented in this contribution allows automatic alignment of histological preparations provided that nuclei or comparable structures extend over several sections and are capable of being extracted using methods of pattern recognition. The method was applied to 50 sections with Nissl staining. The nuclei were extracted from the images and evaluated by application of the algorithm. All image pairs were correctly aligned. An integrated control mechanism ensures automatic detection of incorrectly aligned images. PMID- 15780889 TI - A SAS/IML program for simulating pharmacokinetic data. AB - Data simulation can be an invaluable tool for optimizing the design of bioequivalence trials. It can be particularly useful when exploring alternative approaches for assessing product comparability especially in the context of encountering various complex experimental situations that can occur in veterinary medicine. With this in mind, we designed a novel SAS/IML program to generate pharmacokinetic datasets that reflect the various kinetic, population, and study design characteristics that complicate the bioequivalence evaluation of animal health products. Developing this simulation program within SAS provides an opportunity to utilize the statistical capabilities of this software platform. PMID- 15780891 TI - Augmented convex hull plots: Rationale, implementation in R and biomedical applications. AB - The paper addresses the possibility to replace cluttered multi-group scatter plots with augmented convex hull plots. By replacing scatter-plot points with convex hulls, space is gained for visualization of descriptive statistics with error bars or confidence ellipses within the convex hulls. An informative addition to the plot is calculation of the area of convex hull divided by corresponding group size as a bivariate dispersion measure. Marginal distributions can be depicted on the sides of the main plot in established ways. Bivariate density plots might be used instead of convex hulls in the presence of outliers. Like any scatter-plot type visualization, the technique is not limited to raw data -- points can be derived from any dimension reduction technique, or simple functions can be used as axes instead of original dimensions. The limited possibilities for producing such plots in existing software are surveyed, and our general and flexible implementation in R -- the publicly available chplot function -- is presented. Examples based on our daily biostatistical consulting practice illustrate the technique with various options. PMID- 15780890 TI - MSurvPow: a FORTRAN program to calculate the sample size and power for cluster randomized clinical trials with survival outcomes. AB - Manatunga and Chen [A.K. Manatunga, S. Chen, Sample size estimation for survival outcomes in cluster-randomized studies with small cluster sizes, Biometrics 56 (2000) 616-621] proposed a method to estimate sample size and power for cluster randomized studies where the primary outcome variable was survival time. The sample size formula was constructed by considering a bivariate marginal distribution (Clayton-Oakes model) with univariate exponential marginal distributions. In this paper, a user-friendly FORTRAN 90 program was provided to implement this method and a simple example was used to illustrate the features of the program. PMID- 15780892 TI - Semi-automated image processing system for micro- to macro-scale analysis of immunohistopathology: application to ischemic brain tissue. AB - Immunochemical staining techniques are commonly used to assess neuronal, astrocytic and microglial alterations in experimental neuroscience research, and in particular, are applied to tissues from animals subjected to ischemic stroke. Immunoreactivity of brain sections can be measured from digitized immunohistology slides so that quantitative assessment can be carried out by computer-assisted analysis. Conventional methods of analyzing immunohistology are based on image classification techniques applied to a specific anatomic location at high magnification. Such micro-scale localized image analysis limits one for further correlative studies with other imaging modalities on whole brain sections, which are of particular interest in experimental stroke research. This report presents a semi-automated image analysis method that performs convolution-based image classification on micro-scale images, extracts numerical data representing positive immunoreactivity from the processed micro-scale images and creates a corresponding quantitative macro-scale image. The present method utilizes several image-processing techniques to cope with variances in intensity distribution, as well as artifacts caused by light scattering or heterogeneity of antigen expression, which are commonly encountered in immunohistology. Micro-scale images are composed by a tiling function in a mosaic manner. Image classification is accomplished by the K-means clustering method at the relatively low-magnification micro-scale level in order to increase computation efficiency. The quantitative macro-scale image is suitable for correlative analysis with other imaging modalities. This method was applied to different immunostaining antibodies, such as endothelial barrier antigen (EBA), lectin, and glial fibrillary acidic protein (GFAP), on histology slides from animals subjected to middle cerebral artery occlusion by the intraluminal suture method. Reliability tests show that the results obtained from immunostained images at high magnification and relatively low magnification are virtually the same. PMID- 15780893 TI - [Emergent infectious diseases: importance for public health, epidemiology, promoting factors, and prevention]. AB - At the end of the 70s, it was said that eradicating infectious diseases was possible. With the occurrence of AIDS and the discovery of new pathogens the come back of infectious diseases was noted, 15 to 20 years later, and the concept of emerging infections was defined. An emerging infection is the appearance of a new infection or the increase of its incidence if not new. It also includes infections for which the incidence could increase because of favorable conditions. Several scenarios can be proposed: 1) the appearance of a new infection (AIDS or SARS); 2) the appearance of a known infection where it did not exist previously (West Nile virus infection in the USA); 3) an infection that existed but that was not diagnosed or could not be linked precisely to a particular infectious agent (hepatitis C before the discovery of the hepatitis C virus; 4) a known infection that increases its usual incidence (outbreaks...); 5) infectious diseases for which current conditions may facilitate transmission (decrease of immunization coverage...). The deliberate release of a biological agent is one of the possible scenarios of emergence. Qualitative changes of infectious agents such as the resistance to anti-infectious agents may also contribute, through a selection mechanism, to the emergence of new infectious threat. Emerging infections result from the interaction between the agent, the host, and environment evolving according to human activity. Surveillance, research, laboratory capacity, and an effective public health system are key factors for their control. PMID- 15780894 TI - [Tigecycline: a new antibiotic in ongoing clinical development]. AB - Antimicrobial resistance is an increasing public health concern, all the more as it is concurrently accompanied by limited antimicrobial drugs development, especially broad-spectrum antibiotics. Tigecycline, the first in the novel glycylcycline class to undergo clinical development, showed extensive in vitro activity against a broad range of Gram positive aerobes, among which Staphylococcus aureus, Streptococcus pneumoniae, and Gram negative pathogens, such as Haemophilus influenzae and enterobacteriae, despite these germs resistance to beta-lactams, vancomycin, and a number of other common antibiotics. Tigecycline is also active against some anaerobes. Data obtained in phase II clinical trials have shown good tolerance and efficacy of tigecycline administered by intravenous infusion in the treatment of complicated skin and skin structure infections and intra-abdominal infections. Should these results be confirmed by phase III clinical trials, tigecycline would provide a new option in the ongoing fight against resistant pathogens, which would usefully add to the present antibacterial armamentarium, particularly in severe or hospital treated infections. PMID- 15780895 TI - [Increase of Hantavirus infections in France, 2003]. AB - BACKGROUND: Hantaviral infections causing hemorrhagic fever with renal syndrome are endemic in North Eastern France. Humans are contaminated by the inhalation of aerosols contaminated by rodent faeces. In February 2003, the National reference centre (NRC) for hemorrhagic fevers detected an increased number of cases. An investigation was carried out to confirm the outbreak and take appropriate control measures. METHODS: Cases were collected by the NRC. A case was defined as a person living in France with symptoms compatible with hantaviral infection and a positive blood test both for specific Puumala IgM, and IgG. Clinical information and at-risk exposures during the 2 weeks before onset were recorded. RESULTS: In 2003, 128 cases were diagnosed (61 in 2002). The median age of patients was 38, 77% were men and 82% were hospitalized. Patients were living in North-Eastern France. Clusters were detected in the Ardennes and Oise districts. Occupation (35%) (in agriculture, forestry, and construction work), manipulation of firewood (35%), gardening (29%), and outdoors leisure (14%) were identified as at-risk exposures in these cases. CONCLUSIONS: An increased number of positive diagnoses of hantaviral infections was confirmed. The location and at-risk exposures of the cases were identical in previous investigations. Exclusion and prevention of rodents' access to houses and avoiding the inhalation of contaminated dust are the only possible prevention measures of hantaviral infections. Information about the disease and its prevention needs to be made widely available to both healthcare professionals and the general population living in endemic areas. PMID- 15780896 TI - [Prevalence and risk factors of hepatitis A infection in an HIV-infected French population]. AB - BACKGROUND: There are common risk factors between hepatitis A virus (HAV) and human immuno deficiency virus (HIV) infections. OBJECTIVES: We tried to evaluate if HIV-infected patients could be at risk for HAV. More over, HAV could worsen prognosis of HIV infection and HAV vaccination was then to be considered. Thus we assessed the prevalence and risk factors of HAV infection in an HIV-infected population. PATIENTS AND METHODS: Seroprevalence and risk factors for HAV were studied among 154 HIV-positive patients followed in a Parisian hospital (mean age: 42 years, male patients: 70.8%, female patients: 29.2%). They were screened for HAV antibodies and answered a questionnaire on risk factors for HAV and means of HIV contamination. RESULTS: The global prevalence was 72.7% [IC95%: 65.7 79.7]. We excluded patients who were born in highly endemic areas where seroprevalence reached 60% [IC95%: 51.2-70]. The HAV seroprevalence was almost 100% in migrants from highly endemic countries and for those born before 1946. The multivariate analysis showed that risk factors were the geographic origin [OR=20.88; IC95%: 2.40-181], age [OR = 2.33; IC95%: 1.24-4.39], and hemophilia [OR = 13.78; IC95%: 1.34-141]. CONCLUSION: Our results suggest that a screening test for HAV antibodies should be performed before vaccination, especially in HIV infected patients born after 1946 or in non-endemic countries. PMID- 15780897 TI - [Human immunodeficiency virus type 1 subtypes in cohort of infected patients, Saint-Etienne, France, from 1984 to 2003]. AB - OBJECTIVE: The authors had for aim to study the distribution of HIV-1 subtypes in a cohort of HIV-1 positive patients in the University hospital of Saint-Etienne, France, and to describe the epidemiological characteristics of patients infected with a non-B subtype strain. DESIGN: An epidemiological study was made on 271 HIV 1 positive patients followed up in the Infectious Diseases Department over 20 years. All patients sample were subtyped by serotyping and some samples were also tested by genotyping. RESULTS: Two hundred and sixty-four patients (191 men and 73 women) were found infected by an HIV-1 strain belonging to the M group. After combining serotyping and genotyping results, 195 patients were found infected by a B subtype and 69 by a non-B subtype. Most of the latter strains belonged to an A subtype or related ones. The following factors were shown to be linked to an infection by a non-B strain: being born abroad, having contracted the infection though heterosexual practice, and being a woman. The incidence of non-B strains increased regularly over time (to reach more than 40% in 2003). This progression was especially noted for men born in France with risky sexual behaviour. CONCLUSION: These results indicate that more than 40% of HIV-1 new cases detected in the Saint-Etienne area are related to non-B strains and that strains of A and related subtypes are common in the local population with risky sexual behaviour. PMID- 15780898 TI - [Burkholderia pseudomallei infection after travelling in Southeast Asia, case study]. PMID- 15780899 TI - [Parvovirus B19 infection revealed by prolonged dysesthesia]. PMID- 15780900 TI - [Flexispira rappini bacteremia and common variable immuodeficiency]. PMID- 15780901 TI - [Valganciclovir maintenance therapy in AIDS: treatment failure due to the development of cytomegalovirus resistance to ganciclovir]. PMID- 15780902 TI - [Cervical tuberculosis in children: a six case study]. PMID- 15780903 TI - Antimicrobial use in food animals and human health. PMID- 15780904 TI - Chemoprevention of prostate cancer. AB - Prostate cancer is an attractive target for chemoprevention because of its ubiquity, treatment-related morbidity, long latency between premalignant lesions and clinically evident cancer, and defined molecular pathogenesis. Prevention of this disease would have a major impact on disease-associated cost, morbidity, and mortality for a large segment of the population. A major advance in prevention of prostate cancer came in 2003 with the publication of the Prostate Cancer Prevention Trial (PCPT). This overview summarizes the results of that trial, the design of other large-scale trials, and advances in understanding of the molecular mechanisms underlying the effect of other promising agents, including dutasteride, selenium, Vitamin E, Vitamin D, COX-2 inhibitors, lycopene, and green tea. PMID- 15780905 TI - Overview of monoclonal antibodies in cancer therapy: present and promise. AB - After 30 years of development, therapy with monoclonal antibodies has started to realize its promise. Clinical use is most widespread in the field of oncology, where half of the agents approved for routine clinical use are employed and a large number of molecules are currently undergoing clinical trials. In the past 2 years alone, three new compounds-the radiolabeled antibody (131)I-tositumomab and two antibodies targeting growth factor receptors (bevacizumab and cetuximab)-have received FDA approval for indications in oncology. This review summarizes the development of this exciting treatment modality over the last three decades, examines the outcome of treatment with these new antibodies and others available for routine clinical use (i.e. rituximab, trastuzumab, alemtuzumab, gemtuzumab ozogamicin, (90)Y-ibritumomab tiuxetan) in standard indications and in experimental settings, and gives a brief outlook on possible future developments. PMID- 15780906 TI - Mechanisms of delivery of ubiquitylated proteins to the proteasome: new target for anti-cancer therapy? AB - The proteasome is the main proteolytic machinery of the cell. It is responsible for the basal turnover of many intracellular polypeptides, the elimination of abnormal proteins and the generation of the vast majority of peptides presented by class I major histocompatibility complex molecules. Proteasomal proteolysis is also involved in the control of virtually all cellular functions and major decisions through the spatially and timely regulated destruction of essential cell regulators. Therefore, the elucidation of its molecular mechanisms is crucial for the full understanding of the physiology of cells and whole organisms. Conversely, it is increasingly clear that proteasomal degradation is either altered in numerous pathological situations, including many cancers and diseases resulting from aberrant cell differentiation, or instrumental for the development of these pathologies. This, consequently, makes it an attractive target for therapeutical intervention. There is ample evidence that most cell proteins must be polyubiquitylated prior to proteasomal degradation. If the structure and the mode of functioning of the proteasome, as well as the enzymology of ubiquitylation, are relatively well understood, how substrates are delivered to and recognized by the proteolytic machine has remained mysterious till recently. The recent literature indicates that the mechanisms involved are multiple, complex and exquisitely regulated and provides new potential targets for anti-cancer pharmacological intervention. PMID- 15780907 TI - Molecular mechanisms of tumor vascularization. AB - Tumor angiogenesis is a fast growing sub-domain of angiogenesis research and tumor biology. Basic mechanisms have been unraveled and many key players identified. For many years, tumor vascularization was explained solely by the ingrowth of new vessels into the tumor from preexisting one's. However, in recent years, additional mechanisms have been recognized. These include angioblasts recruitment, cooption, vasculogenic mimicry and mosaic vessels. These different mechanisms may exist concomitantly in the same tumor or may be selectively involved in a specific tumor type or host environment. In this article, we will review, in depth, these different mechanisms and also discuss some aspects of anti-angiogenic tumor therapy. PMID- 15780908 TI - Role of c-Kit and erythropoietin receptor in erythropoiesis. AB - Erythropoiesis is regulated by a number of growth factors, among which stem cell factor (SCF) and erythropoietin (Epo) play a non-redundant function. Viable mice with mutations in the SCF gene (encoded by the Steel (Sl) locus), or its receptor gene c-Kit (encoded by the White spotting (W) locus) develop a hypoplastic macrocytic anemia. Mutants of W or Sl that are completely devoid of c-Kit or SCF expression die in utero of anemia between days 14 and 16 of gestation and contain reduced numbers of erythroid progenitors in the fetal liver. Likewise, Epo and Epo receptor (Epo-R)-deficient mice die in utero due to a marked reduction in the number of committed fetal liver derived erythroid progenitors. Thus, committed erythroid progenitors require both c-Kit and Epo-R signal transduction pathways for their survival, proliferation and differentiation. In vitro, Epo alone is capable of generating mature erythroid progenitors; however, a combined treatment of Epo and SCF results in synergistic proliferation and expansion of developing erythroid progenitors. This review summarizes recent advances made towards understanding the signaling mechanisms by which Epo-R and c-Kit regulate growth, survival, and differentiation of erythroid progenitors alone and cooperatively. PMID- 15780909 TI - Soft tissue sarcoma: why not treated? AB - BACKGROUND: Soft tissue sarcomas (STS) are uncommon malignancies and elderly STS patients have been reported to receive less definitive treatment compared to young STS patients. The present study was performed to investigate whether withholding treatment was based on disease specific aspects, patients' general health condition, comorbidity or a combination of these. METHODS: Patients with primary STS, registered by the Comprehensive Cancer Center North-Netherlands (CCCN) from 1989 to 1999, were analyzed retrospectively with regard to the inclusion-criteria: no primary anti-tumor treatment. RESULTS: From 1989 to 1999, 620 patients (including 56 Kaposi sarcoma) were registered with primary STS. Seventy-six patients (13%) were registered as untreated. Nineteen patients were excluded. Records of 57 patients, median age 71 years (range 23-92, 40 patients > or =65 years, 17 patients < 65 years) were examined. The reasons for no treatment were irresectability of the sarcoma (65%), metastatic disease (11%), comorbidity (4%), poor general health (5%), death prior to therapy (7%) and refusal of therapy (3%) (motivation not documented in 5%). CONCLUSIONS: Thirteen percent of all STS patients within the CCCN region were not treated, 70% of these patients were elderly. Withholding treatment was mostly disease-related (76%), e.g. irresectable retroperitoneal STS or metastatic disease; for 19% of the patients, it was related to their poor general health. The decision to refrain from cancer treatment was justifiable in all these STS patients. PMID- 15780911 TI - The Gunther Laukien Prize. AB - The Gunther Laukien Prize, established in 1999, intends to recognize recent cutting-edge experimental NMR research with a high probability of enabling beneficial new applications. It is awarded yearly at the Experimental NMR Conference (ENC). PMID- 15780912 TI - NMR studies of protein structure and dynamics. AB - Recent advances in solution NMR spectroscopy have significantly extended the spectrum of problems that can now be addressed with this technology. In particular, studies of proteins with molecular weights on the order of 100 kDa are now possible at a level of detail that was previously reserved for much smaller systems. An example of the sort of information that is now accessible is provided in a study of malate synthase G, a 723 residue enzyme that has been a focal point of research efforts in my laboratory. Details of the labeling schemes that have been employed and optimal experiments for extraction of structural and dynamics information on this protein are described. NMR studies of protein dynamics, in principle, give insight into the relation between motion and function. A description of deuterium-based spin relaxation methods for the investigation of side chain dynamics is provided. Examples where millisecond (ms) time scale dynamics play an important role and where relaxation dispersion NMR spectroscopy has been particularly informative, including applications involving the membrane enzyme PagP and mutants of the Fyn SH3 domain that fold on a ms time scale, are presented. PMID- 15780913 TI - Effects of T2-relaxation in MAS NMR spectra of the satellite transitions for quadrupolar nuclei: a 27Al MAS and single-crystal NMR study of alum KAl(SO4)2.12H2O. AB - Asymmetries in the manifold of spinning sidebands (ssbs) from the satellite transitions have been observed in variable-temperature 27Al MAS NMR spectra of alum (KAl(SO4)2.12H2O), recorded in the temperature range from -76 to 92 degrees C. The asymmetries decrease with increasing temperature and reflect the fact that the ssbs exhibit systematically different linewidths for different spectral regions of the manifold. From spin-echo 27Al NMR experiments on a single-crystal of alum, it is demonstrated that these variations in linewidth originate from differences in transverse (T2) relaxation times for the two inner (m=1/2<-->m=3/2 and m=-1/2<-->m=-3/2) and correspondingly for the two outer (m=3/2<-->m=5/2 and m=-3/2<-->m=-5/2) satellite transitions. T2 relaxation times in the range 0.5-3.5 ms are observed for the individual satellite transitions at -50 degrees C and 7.05 T, whereas the corresponding T1 relaxation times, determined from similar saturation-recovery 27Al NMR experiments, are almost constant (T1=0.07-0.10 s) for the individual satellite transitions. The variation in T2 values for the individual 27Al satellite transitions for alum is justified by a simple theoretical approach which considers the cross-correlation of the local fluctuating fields from the quadrupolar coupling and the heteronuclear (27Al-1H) dipolar interaction on the T2 relaxation times for the individual transitions. This approach and the observed differences in T2 values indicate that a single random motional process modulates both the quadrupolar and heteronuclear dipolar interactions for 27Al in alum at low temperatures. PMID- 15780914 TI - The use of multivariate MR imaging intensities versus metabolic data from MR spectroscopic imaging for brain tumour classification. AB - This study investigated the value of information from both magnetic resonance imaging and magnetic resonance spectroscopic imaging (MRSI) to automated discrimination of brain tumours. The influence of imaging intensities and metabolic data was tested by comparing the use of MR spectra from MRSI, MR imaging intensities, peak integration values obtained from the MR spectra and a combination of the latter two. Three classification techniques were objectively compared: linear discriminant analysis, least squares support vector machines (LS SVM) with a linear kernel as linear techniques and LS-SVM with radial basis function kernel as a nonlinear technique. Classifiers were evaluated over 100 stratified random splittings of the dataset into training and test sets. The area under the receiver operating characteristic (ROC) curve (AUC) was used as a global performance measure on test data. In general, all techniques obtained a high performance when using peak integration values with or without MR imaging intensities. For example for low- versus high-grade tumours, low- versus high grade gliomas and gliomas versus meningiomas, the mean test AUC was higher than 0.91, 0.94, and 0.99, respectively, when both MR imaging intensities and peak integration values were used. The use of metabolic data from MRSI significantly improved automated classification of brain tumour types compared to the use of MR imaging intensities solely. PMID- 15780915 TI - Pattern pulses: design of arbitrary excitation profiles as a function of pulse amplitude and offset. AB - A novel class of pulses is presented which can be regarded as a generalization of both frequency-selective pulses and B1-selective pulses. The excitation profile of these pulses forms a pre-defined pattern in two dimensions, which are spanned by pulse offset and radio-frequency (RF) amplitude. The presented pulses were designed numerically based on principles of optimal control theory. For simple test patterns, we demonstrate the flexibility of this approach by simulations and experiments. This previously unknown flexibility may trigger novel applications in NMR spectroscopy and imaging. As a first practical application, we demonstrate a direct approach for calibrating RF pulses. PMID- 15780916 TI - Algebraic description of spin 3/2 dynamics in NMR experiments. AB - The dynamics of spin 3/2 systems is analyzed using the density matrix theory of relaxation. By using the superoperator formalism, an algebraic formulation of the density matrix's evolution is obtained, in which the contributions from free relaxation and RF application are easily factored out. As an intermediate step, an exact form for the propagator of the density matrix for a spin 3/2 system, in the presence of static quadrupolar coupling, inhomogeneous static magnetic field, and relaxation is demonstrated. Using this algebraic formulation, exact expressions for the behavior of the density matrix in the classical one-, two-, and three-pulse experiments are derived. These theoretical formulas are then used to illustrate the bias introduced on the measured relaxation parameters by the presence of large spatial variations in the B0 and B1 fields. The theoretical predictions are easily evaluated through simple matrix algebra and the results agree very well with the experimental observations. This approach could prove useful for the characterization of the spatial variations of the signal intensity in multiple quantum-filtered sodium MRI experiments. PMID- 15780917 TI - Velocity imaging by ex situ NMR. AB - A pulsed field gradient stimulated spin-echo NMR sequence is combined with imaging methods to spatially resolve velocity distributions and to measure 2D velocity maps ex situ. The implementation of these techniques in open sensors provides a powerful non-invasive tool to measure molecular displacement in a large number of applications inaccessible to conventional closed magnets. The method is implemented on an open tomograph that provides 3D spatial localization by combining slice selection in the presence of a uniform static magnetic field gradient along the depth direction with pulsed field gradients along the two lateral directions. Different pipe geometries are used to demonstrate that the sequence performs well even in the extremely inhomogeneous B0 and B1 fields of these sensors. PMID- 15780918 TI - Triple-quantum dynamics in multiple-spin systems undergoing magic-angle spinning: application to 13C homonuclear correlation spectroscopy. AB - We analyze the multiple-quantum dynamics governed by a new homonuclear recoupling strategy effecting an average dipolar Hamiltonian comprising three-spin triple quantum operators (e.g., S(p)+S(q)+S(r)+) under magic-angle spinning conditions. Analytical expressions are presented for polarization transfer processes in systems of three and four coupled spins-1/2 subject to triple-quantum filtration (3QF), and high-order multiple-quantum excitation is investigated numerically in moderately large clusters, comprising up to seven spins. This recoupling approach gives highly efficient excitation of triple-quantum coherences: ideally, up to 67% of the initial polarization may be recovered by 3QF in three-spin systems in polycrystalline powders. Two homonuclear 2D correlation strategies are demonstrated experimentally on powders of uniformly 13C-labeled alanine and tyrosine: the first correlates the single-quantum spectrum in the first dimension with the corresponding 3QF spectrum along the other. The second protocol correlates triple-quantum coherences with their corresponding single-quantum coherences within triplets of coupled spins. PMID- 15780919 TI - Using wavelet de-noised spectra in NMR screening. AB - Principal component analysis (PCA) is a commonly used algorithm in multivariate analysis of NMR screening data. PCA substantially reduces the complexity of data in which a large number of variables are interrelated. For series of NMR spectra obtained for ligand binding, it is commonly used to visually group spectra with a similar response to ligand binding. A series of filters are applied to the experimental data to obtain suitable descriptors for PCA which optimize computational efficiency and minimize the weight of small chemical shift variations. The most common filter is bucketing where adjacent points are summed to a bucket. To overcome some inherent disadvantages of the bucketing procedure we have explored the effect of wavelet de-noising on multivariate analysis, using a series of HSQC spectra of proteins with different ligands present. The combination of wavelet de-noising and PCA is most efficient when PCA is applied to wavelet coefficients. This new algorithm yields good clustering and can be applied to series of one- or two-dimensional spectra. PMID- 15780920 TI - ESR spectrometer with a loop-gap resonator for cw and time resolved studies in a superconducting magnet. AB - The design and performance of an electron spin resonance spectrometer operating at 3 and 9 GHz microwave frequencies combined with a 9-T superconducting magnet are described. The probehead contains a compact two-loop, one gap resonator, and is inside the variable temperature insert of the magnet enabling measurements in the 0-9T magnetic field and 1.5-400 K temperature range. The spectrometer allows studies on systems where resonance occurs at fields far above the g approximately 2 paramagnetic condition such as in antiferromagnets. The low quality factor of the resonator allows time resolved experiments such as, e.g., longitudinally detected ESR. We demonstrate the performance of the spectrometer on the NaNiO2 antiferromagnet, the MgB2 superconductor, and the RbC60 conducting alkaline fulleride polymer. PMID- 15780921 TI - MAS NMR with and without double-quantum filtration at and near the n=0 rotational resonance condition. AB - Spectral lineshapes of MAS NMR spectra of dipolar (re)coupled spin pairs exhibiting considerable chemical shielding anisotropies at and near the so-called n=0 rotational resonance (R2) condition are considered. The n=0 R2 condition is found to be not extremely sharp. Anisotropic interaction parameters such as chemical shielding tensor orientations and the magnitude of the dipolar coupling constant remain sensitively encoded in such lineshapes even when differences in isotropic chemical shielding values of up to 400 Hz (corresponding to ca. half the size of the dipolar coupling constant) are present. Additional double-quantum filtration (DQF) may enhance the sensitivity of spectral lineshapes to anisotropic interaction parameters for even larger differences in isotropic chemical shielding values. The dependence of the DQF efficiency on spin-system parameters as well as on external parameters (Larmor and MAS frequencies) is investigated. Away from R2 conditions a trend to lower DQF efficiencies is found whereas some spin-system parameters are more sensitively encoded in the corresponding spectral lineshapes. Our study is based on numerical simulations, with the known parameters of the 31P spin pair in Na4P2O7.10H2O representing our model case. PMID- 15780922 TI - IFSERF, an isotope-filtered SERF experiment for the precise measurement of proton proton coupling constants between chemically equivalent protons. AB - An isotope-filtered selective refocusing (IFSERF) experiment is presented for the sensitive and precise measurement of the proton-proton coupling constant between chemically equivalent protons. The 2D NMR method combines an initial doubly selective isotope filter based on heteronuclear cross-polarization followed by a selective J-resolved block. The coupling topologies obtained from several 2D variants of the IFSERF experiment are described for the simultaneous measurement of both proton-proton and proton-carbon coupling constants in the involved AA'XX' spin system. Application on the determination of the relative configuration of double bonds in symmetrical molecules is illustrated. PMID- 15780923 TI - The fumarate sensor DcuS: progress in rapid protein fold elucidation by combining protein structure prediction methods with NMR spectroscopy. AB - We illustrate how moderate resolution protein structures can be rapidly obtained by interlinking computational prediction methodologies with un- or partially assigned NMR data. To facilitate the application of our recently described method of ranking and subsequent refining alternative structural models using unassigned NMR data [Proc. Natl. Acad. Sci. USA 100 (2003) 15404] for such "structural genomics"-type experiments it is combined with protein models from several prediction techniques, enhanced to utilize partial assignments, and applied on a protein with an unknown structure and fold. From the original NMR spectra obtained for the 140 residue fumarate sensor DcuS, 1100 1H, 13C, and 15N chemical shift signals, 3000 1H-1H NOESY cross peak intensities, and 209 backbone residual dipolar couplings were extracted and used to rank models produced by de novo structure prediction and comparative modeling methods. The ranking proceeds in two steps: first, an optimal assignment of the NMR peaks to atoms is found for each model independently, and second, the models are ranked based on the consistency between the NMR data and the model assuming these optimal assignments. The low-resolution model selected using this ranking procedure had the correct overall fold and a global backbone RMSD of 6.0 angstrom, and was subsequently refined to 3.7 angstrom RMSD. With the incorporation of a small number of NOE and residual dipolar coupling constraints available very early in the traditional spectral assignment process, a model with an RMSD of 2.8 angstrom could rapidly be built. The ability to generate moderate resolution models within days of NMR data collection should facilitate large scale NMR structure determination efforts. PMID- 15780924 TI - Fast multidimensional NMR: radial sampling of evolution space. AB - Multidimensional NMR spectroscopy can be speeded up by limited radial sampling of the time-domain evolution data. The resulting frequency-domain projections are used to reconstruct the full NMR spectrum. New algorithms are proposed to suppress back-projection artifacts while retaining optimum sensitivity. The method is illustrated by experiments on the 900 MHz HNCO spectrum of a protein, HasA. PMID- 15780926 TI - Protein backbone dynamics from N-HN dipolar couplings in partially aligned systems: a comparison of motional models in the presence of structural noise. AB - Residual dipolar couplings (RDCs) provide excellent probes for the exploration of dynamics in biomolecules on biologically relevant time-scales. Applying geometric motional models in combination with high-resolution structures to fit experimental RDCs allows the extraction of local dynamic amplitudes of peptide planes in proteins using only a limited number of data points. Here we compare the behaviour of three simple and intuitive dynamic modes: the Gaussian axial fluctuation model (1D-GAF), the two-site jump model, and a model supposing axially symmetric motion about a mean orientation. The requirement of a structural model makes this kind of methodology potentially very sensitive to structural imprecision. Numerical simulations of RDC dynamic averaging under different regimes show that the anisotropic motional models are more geometrically stringent than the axially symmetric model making it more difficult to alias structural noise as artificial dynamic amplitudes. Indeed, it appears that the model extracts accurate motional amplitudes even in the presence of significant structural error. We also show that a two-site jump model, also assuming the (alpha)C(i-1)-(alpha)C(i) as rotation axis, can only be distinguished from the previously developed GAF model beyond amplitude/jumps of around 40 degrees. The importance of appropriate estimation of the molecular alignment tensor for determination of local motional amplitudes is also illustrated here. We demonstrate a systematic scaling of extracted dynamic amplitudes if a static structure is assumed when determining the alignment tensor from dynamically averaged RDCs. As an example an artificial increase of 0.14 (0.85 compared to the expected 0.71) is observed in the extracted S2 if a pervasive 20 degrees GAF motion is present that is ignored in the tensor determination. Finally we apply a combined approach using the most appropriate motional model, to complete the analysis of dynamic motions from protein G. PMID- 15780925 TI - 15N and 31P solid-state NMR study of transmembrane domain alignment of M2 protein of influenza A virus in hydrated cylindrical lipid bilayers confined to anodic aluminum oxide nanopores. AB - This communication reports the first example of a high resolution solid-state 15N 2D PISEMA NMR spectrum of a transmembrane peptide aligned using hydrated cylindrical lipid bilayers formed inside nanoporous anodic aluminum oxide (AAO) substrates. The transmembrane domain SSDPLVVA(A-15N)SIIGILHLILWILDRL of M2 protein from influenza A virus was reconstituted in hydrated 1,2-dimyristoyl-sn glycero-3-phosphatidylcholine bilayers that were macroscopically aligned by a conventional micro slide glass support or by the AAO nanoporous substrate. 15N and 31P NMR spectra demonstrate that both the phospholipids and the protein transmembrane domain are uniformly aligned in the nanopores. Importantly, nanoporous AAO substrates may offer several advantages for membrane protein alignment in solid-state NMR studies compared to conventional methods. Specifically, higher thermal conductivity of aluminum oxide is expected to suppress thermal gradients associated with inhomogeneous radio frequency heating. Another important advantage of the nanoporous AAO substrate is its excellent accessibility to the bilayer surface for exposure to solute molecules. Such high accessibility achieved through the substrate nanochannel network could facilitate a wide range of structure-function studies of membrane proteins by solid-state NMR. PMID- 15780927 TI - Improving NMR sensitivity in room temperature and cooled probes with dipolar ions. AB - The response of inverse triple resonance cold and conventional probes to ionic strength has been compared under a variety of conditions relevant to protein NMR. Increasing the salt concentration degrades probe performance in terms of sensitivity, and the effect is more severe for cold probes and with increasing magnetic field strength. This is especially noticeable for experiments that involve a spin lock or decoupling, where sensitivity losses compared with pure water can be more than 2-fold. We have investigated the use of glycine as a substitute for salt as a supporting solute for proteins, and we show that it has a minimal effect on probe tuning or performance. Readily available d5-Gly is a useful co-solute for protein NMR, especially at high magnetic field strengths and on cold probes, as it maintains solubility while not degrading probe performance. PMID- 15780928 TI - Interlaced Fourier transformation of ultrafast 2D NMR data. AB - A new protocol for processing the data arising in ultrafast 2D NMR is discussed and exemplified, based on the interlaced Fourier transformation. This approach is capable of dealing in a single, combined fashion, with the two mirror-imaged interferograms arising in this kind of experiment as a result of the acquisition of a train of magnetic field gradient echoes. By combining all the acquired data points into a common Fourier processing procedure the spectral width along the direct-acquisition domain becomes effectively doubled, giving the opportunity of employing acquisition gradients that are approximately half as strong as hitherto required. This in turn should lead to an overall enhancement in the signal-to noise ratio of the experiment of ca. 2, as well as to improvements in the achievable digital resolution. These expectations were tested by carrying out a series of homo- and heteronuclear ultrafast 2D NMR acquisitions, and found systematically fulfilled. The robustness and conditions that allow the interlaced numerical procedure to be implemented in routine analytical applications were explored and are briefly discussed. PMID- 15780929 TI - Filling the gaps in replication restart pathways. AB - Recent work by clarifies two mechanisms of replication fork assembly in E. coli: in vitro, the presence of a leading nascent strand dictates a PriA/B DnaT dependent mechanism, whereas a gapped fork specifies PriC-mediated restart. PMID- 15780930 TI - Transcriptional activation: mediator can act after preinitiation complex formation. AB - The Mediator complex connects transcriptional activators bound at enhancers with RNA polymerase (Pol) II. demonstrate that Mediator also has an important role in transcriptional activation after recruitment of the Pol II machinery to promoters. PMID- 15780931 TI - Finding the right template: RNA Pol IV, a plant-specific RNA polymerase. AB - A paradox of RNA-directed transcriptional silencing is that low-level transcription through the silenced locus is observed and may help maintain the silenced state. Discovery of a plant-specific RNA polymerase that directs heterochromatic silencing helps to resolve this issue. PMID- 15780932 TI - Translational complexity of the fragile x mental retardation protein: insights from the fly. AB - Through the awesome power of Drosophila genetics, two recent studies reveal novel mechanisms by which the Fragile X Mental Retardation protein regulates selective mRNA translation, controlling key steps of germline development during oogenesis and neuronal development during synaptogenesis. PMID- 15780933 TI - Structural bases of unphosphorylated STAT1 association and receptor binding. AB - The crystal structure has been determined at 3.0 A resolution for an unphosphorylated STAT1 (1-683) complexed with a phosphopeptide derived from the alpha chain of interferon gamma (IFNgamma) receptor. Two dimer interfaces are seen, one between the N domains (NDs) (amino acid residues 1-123) and the other between the core fragments (CFs) (residues 132-683). Analyses of the wild-type (wt) and mutant STAT1 proteins by static light scattering, analytical ultracentrifugation, and coimmunoprecipitation suggest that STAT1 is predominantly dimeric prior to activation, and the dimer is mediated by the ND interactions. The connecting region between the ND and the CF is flexible and allows two interconvertable orientations of the CFs, termed "antiparallel" or "parallel," as determined by SH2 domain orientations. Functional implications of these dimer conformations are discussed. Also revealed in this structure is the detailed interaction between STAT1 SH2 domain and its docking site on IFNgamma receptor. PMID- 15780934 TI - Defining a centromere-like element in Bacillus subtilis by Identifying the binding sites for the chromosome-anchoring protein RacA. AB - Chromosome segregation during sporulation in Bacillus subtilis involves the anchoring of sister chromosomes to opposite ends of the cell. Anchoring is mediated by RacA, which acts as a bridge between a centromere-like element in the vicinity of the origin of replication and the cell pole. To define this element we mapped RacA binding sites by performing chromatin immunoprecipitation in conjunction with gene microarray analysis. RacA preferentially bound to 25 regions spread over 612 kb across the origin portion of the chromosome. Computational and biochemical analysis identified a GC-rich, inverted 14 bp repeat as the recognition sequence. Experiments with single molecules of DNA demonstrated that RacA can condense nonspecific DNA dramatically against appreciable forces to form a highly stable protein-DNA complex. We propose that interactions between DNA bound RacA molecules cause the centromere-like element to fold up into a higher order complex that fastens the chromosome to the cell pole. PMID- 15780935 TI - Fusion of hairpin telomeres by the B. burgdorferi telomere resolvase ResT implications for shaping a genome in flux. AB - Spirochetes of the genus Borrelia include the causative agents of Lyme disease and relapsing fever. These bacteria have a highly segmented genome where most replicons are linear molecules terminated by covalently closed hairpin telomeres. Moreover, these genomes appear to be in a state of flux with extensive and ongoing DNA rearrangements by unknown mechanisms. The B. burgdorferi telomere resolvase ResT generates the hairpin telomeres from replication intermediates in a reaction with mechanistic similarities to that catalyzed by type IB topoisomerases and tyrosine recombinases. We report here the unexpected ability of ResT to catalyze the fusion of hairpin telomeres in a reversal of the telomere resolution reaction. We propose that stabilized ResT-mediated telomere fusions are an underlying force for maintaining the B. burgdorferi genome in a state of flux. PMID- 15780936 TI - HIF-1alpha induces genetic instability by transcriptionally downregulating MutSalpha expression. AB - Hypoxia promotes genetic instability by undefined mechanisms. The transcription factor HIF-1alpha is crucial for the cellular response to hypoxia and is frequently overexpressed in human cancers, resulting in the activation of genes essential for cell survival. Here, we demonstrate that HIF-1alpha is responsible for genetic instability at the nucleotide level by inhibiting MSH2 and MSH6, thereby decreasing levels of the MSH2-MSH6 complex, MutSalpha, which recognizes base mismatches. HIF-1alpha displaces the transcriptional activator Myc from Sp1 binding to repress MutSalpha expression in a p53-dependent manner; Sp1 serves as a molecular switch by recruiting HIF-1alpha to the gene promoter under hypoxia. Furthermore, in human sporadic colon cancers, HIF-1alpha overexpression is statistically associated with the loss of MSH2 expression, especially when p53 is immunochemically undetectable. These findings indicate that the regulation of DNA repair is an integral part of the hypoxic response, providing molecular insights into the mechanisms underlying hypoxia-induced genetic instability. PMID- 15780937 TI - Swapping function of two chromatin remodeling complexes. AB - SWI/SNF- and ISWI-based complexes have distinct yet overlapping chromatin remodeling activities in vitro and perform different roles in vivo. This leads to the hypothesis that the distinct remodeling functions of these complexes are specifically required for distinct biological tasks. By creating and characterizing chimeric proteins of BRG1 and SNF2h, the motor proteins of human SWI/SNF- and ISWI-based complexes, respectively, we found that a region that includes the ATPase domain specifies the outcome of the remodeling reaction in vitro. A chimeric protein based on BRG1 but containing the SNF2h ATPase domain formed an intact SWI/SNF complex that remodeled like SNF2h. This altered-function complex was active for remodeling and could stimulate expression from some, but not all, SWI/SNF responsive promoters in vivo. Thus, we were able to separate domains of BRG1 responsible for function from those responsible for SWI/SNF complex formation and demonstrate that remodeling functions are not interchangeable in vivo. PMID- 15780938 TI - Conversion of active promoter-RNA polymerase complexes into inactive promoter bound complexes in E. coli by the transcription effector, ppGpp. AB - Guanosine tetraphosphate (ppGpp) is a signal of nutritional stress that regulates transcription. An RNA polymerase rudder mutant rpoC (Delta 312-315) is found to suppress ppGpp deficiency phenotypes by restoring both negative and positive activities of promoter fusions in vivo, as if ppGpp were present. Measurements of defects in transcription of the PargT tRNA promoter with mutant RNA polymerase reveal that the mutant enzyme quantitatively mimics the presence of added ppGpp. DNaseI footprints and mobility shifts under RNA polymerization conditions reveal that the promoter-specific transcription defect of the mutant enzyme can be ascribed to the presence of inactive dead-end promoter complexes with features similar to those of a stable closed complex. We propose that formation of such inactive complexes represents an alternative explanation of "stringent RNA polymerase" mutant behavior to those currently published, and it represents a newly discovered mode of action of ppGpp. PMID- 15780939 TI - Distinction and relationship between elongation rate and processivity of RNA polymerase II in vivo. AB - A number of proteins and drugs have been implicated in the process of transcriptional elongation by RNA polymerase (Pol) II, but the factors that govern the elongation rate (nucleotide additions per min) and processivity (nucleotide additions per initiation event) in vivo are poorly understood. Here, we show that a mutation in the Rpb2 subunit of Pol II reduces both the elongation rate and processivity in vivo. In contrast, none of the putative elongation factors tested affect the elongation rate, although mutations in the THO complex and in Spt4 significantly reduce processivity. The drugs 6-azauracil and mycophenolic acid reduce both the elongation rate and processivity, and this processivity defect is aggravated by mutations in Spt4, TFIIS, and CTDK-1. Our results suggest that, in vivo, a reduced rate of Pol II elongation leads to premature dissociation along the chromatin template and that Pol II processivity can be uncoupled from elongation rate. PMID- 15780940 TI - Elp1p, the yeast homolog of the FD disease syndrome protein, negatively regulates exocytosis independently of transcriptional elongation. AB - The activation of Rab GTPases is a critical focal point of membrane trafficking events in eukaryotic cells; however, the cellular mechanisms that spatially and temporally regulate this process are poorly understood. Here, we identify a null allele of ELP1 as a suppressor of a mutant in a Rab guanine nucleotide exchange factor Sec2p. Elp1p was previously thought to be involved in transcription elongation as part of the Elongator complex. We show that elp1Delta suppression of sec2(ts) is not a result of reduced transcriptional elongation and that Elp1p physically associates with Sec2p. The Sec2p interaction domain of Elp1p is necessary for both Elp1p function and for the polarized localization of Sec2p. Mutations in human Elp1p (IKAP) are a known cause of familial dysautonomia (FD). Our results raise the possibility that regulation of polarized exocytosis is an evolutionarily conserved function of the entire Elongator complex and that FD results from a dysregulation of neuronal exocytosis. PMID- 15780941 TI - Mechanism of sirtuin inhibition by nicotinamide: altering the NAD(+) cosubstrate specificity of a Sir2 enzyme. AB - Sir2 enzymes form a unique class of NAD(+)-dependent deacetylases required for diverse biological processes, including transcriptional silencing, regulation of apoptosis, fat mobilization, and lifespan regulation. Sir2 activity is regulated by nicotinamide, a noncompetitive inhibitor that promotes a base-exchange reaction at the expense of deacetylation. To elucidate the mechanism of nicotinamide inhibition, we determined ternary complex structures of Sir2 enzymes containing nicotinamide. The structures show that free nicotinamide binds in a conserved pocket that participates in NAD(+) binding and catalysis. Based on our structures, we engineered a mutant that deacetylates peptides by using nicotinic acid adenine dinucleotide (NAAD) as a cosubstrate and is inhibited by nicotinic acid. The characteristics of the altered specificity enzyme establish that Sir2 enzymes contain a single site that participates in catalysis and nicotinamide regulation and provides additional insights into the Sir2 catalytic mechanism. PMID- 15780942 TI - Major conformational change in the complex SF3b upon integration into the spliceosomal U11/U12 di-snRNP as revealed by electron cryomicroscopy. AB - In some eukaryotes, a minor class of introns is removed by the U12-dependent spliceosome, which contains the small nuclear ribonucleoprotein (snRNP) heterodimer U11/U12. The U11/U12 di-snRNP forms a molecular bridge that functionally pairs the intron ends of the pre-mRNA. We have determined the three dimensional (3D) structure of the human U11/U12 di-snRNP by single particle electron cryomicroscopy using angular reconstitution and random conical tilt. SF3b, a heteromeric protein complex functionally important for branch site recognition, was located in the U11/U12 di-snRNP by antibody labeling and by identification of structural domains of SF3b155, SF3b49, and p14. The conformation of SF3b bound to the U11/U12 di-snRNP differs from that of isolated SF3b: upon integration into the di-snRNP, SF3b rearranges into a more open form. The manner in which SF3b is integrated in the U11/U12 di-snRNP has important implications for branch site recognition. Furthermore, a putative model of the pre-mRNA binding to the U11/U12 di-snRNP is proposed. PMID- 15780943 TI - Chromosomal translocation mechanisms at intronic alu elements in mammalian cells. AB - Repetitive elements comprise nearly half of the human genome. Chromosomal rearrangements involving these elements occur in somatic and germline cells and are causative for many diseases. To begin to understand the molecular mechanisms leading to these rearrangements in mammalian cells, we developed an intron-based system to specifically induce chromosomal translocations at Alu elements, the most numerous family of repetitive elements in humans. With this system, we found that when double-strand breaks (DSBs) were introduced adjacent to identical Alu elements, translocations occurred at high frequency and predominantly arose from repair by the single-strand annealing (SSA) pathway (85%). With diverged Alu elements, translocation frequency was unaltered, yet pathway usage shifted such that nonhomologous end joining (NHEJ) predominated as the translocation pathway (93%). These results emphasize the fluidity of mammalian DSB repair pathway usage. The intron-based system is highly adaptable to addressing a number of issues regarding molecular mechanisms of genomic rearrangements in mammalian cells. PMID- 15780944 TI - Estrogen receptor beta: the antimechanostat? AB - We have known for sometime that sex hormones influence the growth, preservation, and loss of bone tissue in the skeleton. However, we are only beginning to recognize how estrogen influences the responsiveness of the skeleton to exercise. Frost's mechanostat theory proposes that estrogen reduces the mechanical strain required to initiate an osteogenic response, but this may only occur at the endocortical and trabecular bone surfaces. The discovery of estrogen receptors alpha and beta may help us to understand the bone surface-specific effects of exercise. Findings from estrogen receptor knockout mice suggest that the activity of ERalpha may explain the positive interaction between estrogen and exercise on bone formation near marrow, that is, endocortical and trabecular bone surfaces. Estrogen inhibits the anabolic exercise response at the periosteal surface, and this we propose is due to the activation of ERbeta. Signaling through this receptor retards periosteal bone formation and suppresses gains in bone size and bone strength, and for these reasons it behaves as an antimechanostat. PMID- 15780945 TI - An orientation distribution function for trabecular bone. AB - We describe a new method for quantifying the orientation of trabecular bone from three-dimensional images. Trabecular lattices from five human vertebrae were decomposed into individual trabecular elements, and the orientation, mass, and thickness of each element were recorded. Continuous functions that described the total mass (M(phi,theta)) and mean thickness (tau(phi,theta)) of all trabeculae as a function of orientation were derived. The results were compared with experimental measurements of the elastic modulus in three principal anatomic directions. A power law scaling relationship between the anisotropies in mass and elastic modulus was observed; the scaling exponent was 1.41 (R2=0.88). As expected, the preponderance of trabecular mass was oriented along the cranial caudal direction; on average, there was 3.4 times more mass oriented vertically than horizontally. Moreover, the vertical trabeculae were 30% thicker, on average, than the horizontal trabeculae. The vertical trabecular thickness was inversely related to connectivity (R2=0.70; P=0.07), suggesting a possible organization into either few, thick trabeculae or many thin trabeculae. The method, which accounts for the mechanical connectedness of the lattice, provides a rapid way to both visualize and quantify the three-dimensional organization of trabecular bone. PMID- 15780946 TI - Retinoid-induced bone thinning is caused by subperiosteal osteoclast activity in adult rodents. AB - Excess of Vitamin A (retinol) and related compounds (retinoids) induces bone fragility and is associated with increased hip fracture incidence in humans. Yet, their impact on the adult skeleton has been studied in relatively little detail. It is assumed that they induce generalized bone loss and decrease long-bone thickness due to reduction of radial bone growth. Here we characterized early skeletal responses of adult rodents to retinoid treatment, revealing novel aspects of retinoid action on the mature skeleton. The retinoid Ro 13-6298, given subcutaneously for 4 days, induced bone loss in the hind limbs of 12- and 56-week old rats and of 15-week-old mice. In vivo monitoring of bone mass and geometry changes by peripheral quantitative computed tomography demonstrated that bone mass decline was due to subperiosteal cortical bone loss, which induced a shrinkage of bone diameter, whilst cancellous bone mass was preserved. We observed that the native retinoic acid isomer all-trans RA induced an identical pattern of bone loss. Histomorphometric evaluation revealed that increased subperiosteal osteoclastic bone resorption caused the cortical bone destruction. Interestingly, bone resorption was suppressed in cancellous bone, which was in agreement with reduced in vitro formation of osteoclasts from bone marrow cells that were derived from the proximity of cancellous bone. The retinoid-induced increase in subperiosteal bone resorption could be blocked by bisphosphonate as direct potent inhibitor of osteoclast action, but not by estradiol. Retinoid treatment induced a reduction of bone-forming surfaces at the subperiosteal site, but not in cancellous bone. In vitro osteoblast performance was also reduced or unchanged, depending on the cellular system used and assay type/duration. In conclusion, our studies revealed that the impact of retinoids on bone is highly bone-compartment-specific at early treatment phases. Furthermore, we showed that bone diameter shrinks in the adult skeleton after retinoid treatment due to subperiosteal osteoclastic bone resorption. Thus, retinoid-induced bone thinning is not only due to reduced radial bone growth as previously assumed. Our findings might explain why high intake of retinol is associated with increased hip fracture risk in the elderly and suggest a therapy to prevent such potential negative effects. PMID- 15780947 TI - Loss of the transcription factor p45 NF-E2 results in a developmental arrest of megakaryocyte differentiation and the onset of a high bone mass phenotype. AB - NF-E2 is a transcription factor required for megakaryocyte differentiation. The phenotype of mice deficient in p45 NF-E2 has been characterized by increased numbers of immature megakaryocytes and the absence of functional platelets. These mice also exhibited a high bone mass phenotype with up to a 6-fold increase in trabecular bone volume and a 3- to 5-fold increase in the bone formation rate. Our data indicated that both osteoblast and osteoclast numbers were increased in vivo with a 4- to 10-fold increase in osteoblast number/tissue area and approximately a 5-fold increase in osteoclast number/tissue area. Serum osteocalcin levels were also increased in NF-E2-deficient mice, corroborating the histomorphometric data and confirming that the osteoblasts were functional. Urinary cross-links levels were measured to confirm osteoclast activity. Interestingly, the increased bone was observed only in bony sites of hematopoiesis, and was not seen in flat bones such as calvariae. We showed that cells of the osteoblast lineage do not express NF-E2 mRNA. The increased bone phenotype was adoptively transferred into irradiated wild-type mice using spleen cells from NF-E2-deficient mice. These observations suggest that a megakaryocyte osteoblast interaction occurs which is anabolic for bone. PMID- 15780948 TI - Osteoblast-specific expression of insulin-like growth factor-1 in bone of transgenic mice induces insulin-like growth factor binding protein-5. AB - The activities of insulin-like growth factors (IGFs) in bone are modulated by a family of binding proteins (IGFBPs) whose physiological roles remain poorly understood. We have previously shown that targeted overexpression of IGF-I in osteoblasts of transgenic (OC-IGF-I) mice stimulates bone formation. In this model, bone formation is markedly but transiently increased in an age-dependent manner, raising the possibility that IGF-I may be influencing IGFBPs to in turn modulate its paracrine actions within bone. We sought to characterize the IGFBPs in normal mouse bone during development and to determine whether osteoblast targeted overexpression of IGF-I influenced bone IGFBP abundance in vivo. Femoral bone IGFBP content was assessed in control nontransgenic and OC-IGF-I mice by I125-IGF-I ligand and immunoblotting. Bone IGFBP-5 and IGF-I mRNA abundance was determined using real-time reverse transcription (RT)-PCR. Ligand blot of bone extract showed a 30-kDa band, identified as IGFBP-5 by immunoblot, predominated. The abundance of IGFBP-5 declined with age in both control and transgenic bone. Ligand and immunoblot analysis revealed a 5-fold increase in IGFBP-5 protein levels at 3 weeks in transgenic bone (P<0.0001). The elevated IGFBP-5 protein levels were associated with a similar increase in IGF-I mRNA abundance (4-fold, P<0.01) and a significant increase in IGFBP-5 mRNA abundance (1.5-fold). Despite the age-related decline at 6 weeks, IGFBP-5 remained significantly (P<0.01) more abundant in transgenic bone compared to controls. In contrast, bone IGFBP-4 abundance was relatively unchanged by either age or IGF-I overexpression. These studies demonstrate a distinctive developmental pattern of IGFBP-5 content in mouse bone and show that osteoblast-derived IGF-I determines skeletal IGFBP-5 abundance, at least in part by inducing its synthesis. In that IGFBP-5 is thought to stimulate bone formation, directly or via IGF-I action, such changes in bone IGFBP-5 may be important to ensure robust bone acquisition in the early postnatal period. PMID- 15780949 TI - Beta ig-h3 mediates osteoblast adhesion and inhibits differentiation. AB - betaig-h3 is an extracellular matrix (ECM) protein induced by TGF-beta, and it has motifs interacting with the alpha3beta1, alphavbeta5, and alphavbeta3 integrins. Our previous study shows the role of betaig-h3 in osteoblast differentiation and its involvement in melorheostosis, a rare bone disease. Here we demonstrate that betaig-h3 expression is down-regulated during the early stage of differentiation of the murine preosteoblastic cell line, KS483. The recombinant betaig-h3 and its FAS1 domain significantly inhibited in vitro osteoblast differentiation as evaluated by matrix mineralization/bone nodule formation. Furthermore, inhibition of expression of osteoblast differentiation marker genes [such as type I collagen, alkaline phosphatase, and osteocalcin (OC)] was accompanied by suppression of osteoblast-specific transcription factors, Cbfa1/Runx2 and osterix. Flow cytometric analyses, cell adhesion, and inhibition assays disclosed alphavbeta3 and alphavbeta5 as the principal integrins mediating the adhesion of osteoblastic cells to betaig-h3. The disruption of interactions between betaig-h3 and osteoblasts by a function blocking antibody specific for alphavbeta3 but not for alphavbeta5 abolished the inhibitory effect of betaig-h3 on osteoblast differentiation. We suggest that these interacting integrins may play an important role in betaig-h3-mediated inhibition of osteoblast differentiation. PMID- 15780950 TI - Characterisation of cytosolic FK506 binding protein 12 and its role in modulating expression of Cbfa1 and osterix in ROS 17/2.8 cells. AB - FK506 is a commonly used immunosuppressant that mediates its action by exclusively interacting with the cytosolic immunophilin, FK506 binding protein 12 (FKBP12). Although FK506-induced acute osteoporosis is now well recognised, its precise mode of action in osteoblasts remains unclear. Therefore, in the present study we characterised FKBP12 in osteoblasts and investigated the role of FK506 in modulating osteoblast-specific transcription factors, core-binding factor alpha1 (Cbfa1) and osterix gene expression in ROS 17/2.8 cells. RT-PCR, immunolocalisation and Western blotting studies were employed to identify and characterise FKBP12 in rat primary osteoblasts and osteoblast-like osteosarcoma ROS 17/2.8 cells. Western blotting extracts of these cells revealed the 12 kDa and hitherto unreported 10 kDa FKBP isoform that were immunolocalised predominantly to the cytosol. The transient exposure of ROS 17/2.8 cells to H2O2 (100 microM) was found to elevate FKBP12 mRNA after 10 min and protein expression after 24 h. Both PTH (10(-9) M) and 1,25 (OH)2D3 (Vitamin D3) (10(-7) M) suppressed FKBP12 protein expression. FK506 in the therapeutic range (25 nmol/L) suppressed expression of Cbfa1 and osterix mRNA. The inhibition of Cbfa1 isoforms II/III expression was evident at 30 min and the extent of inhibition was sustained at 6 h. Osterix inhibition was also seen after 30 min, however, it became maximal after 6 h. The dose-dependant inhibition of osterix in these cells, carried out using 1.25, 12.5 and 125 nmol/L of FK506 was maximal at 1.25 nmol/L. Cbfa1 isoforms II/III were also maximally inhibited at 1.25 nmol/L; interestingly, the inhibition became less marked at higher concentrations of FK506. Similar dose of FK506 was found to inhibit ROS 17/2.8 cell proliferation; the inhibitory effect however was greater in insulin-stimulated cells. The results of this study suggest that immunosuppressant-induced osteoporosis, which is known to involve accelerated bone resorption by increase in osteoclastogenesis, may in fact also be accentuated by the inhibition of osteoblast differentiation and function. PMID- 15780951 TI - Effects of FGF-2/-9 in calvarial bone cell cultures: differentiation stage dependent mitogenic effect, inverse regulation of BMP-2 and noggin, and enhancement of osteogenic potential. AB - Systemically administered fibroblast growth factors (FGFs) show anabolic effects on bone formation in animals, whereas in vitro cell culture studies have demonstrated that FGFs block mineralized bone nodule formation. These apparently contradictory outcomes indicate that the nature of FGF action is complex and that the biological effect of FGFs may depend on the differentiation stage of osteoblasts, interaction with other cytokines, or the length and mode of exposure to factors. Thus, we have utilized primary calvarial bone cell populations at different maturation phases to determine their responses to 2, FGF-9, and BMP-2, the factors expressed in bone. FGF-2 and FGF-9 stimulated proliferation of the cell populations consisting of more mature osteoblasts, but not those with undifferentiated precursor cells. Continuous treatment with FGF-2/-9 inhibited expression of several osteoblast marker genes and mineralization. However, brief pretreatment with FGF-2/-9 or sequential treatment with FGF-2/-9 followed by BMP 2 led to marked stimulation of mineralization, suggesting that FGFs enhance the intrinsic osteogenic potential. Furthermore, FGF-2 and FGF-9 increased expression of other osteogenic factors BMP-2 and TGFbeta-1. Meanwhile, blocking endogenous FGF signaling, using a virally transduced dominant-negative FGF receptor (FgfR), resulted in drastically reduced expression of the BMP-2 gene, demonstrating for the first time that endogenous FGF/FgfR signaling is a positive upstream regulator of the BMP-2 gene in calvarial osteoblasts. In contrast, expression of a BMP antagonist noggin was inhibited by FGF-2 and FGF-9. Thus, collective data from this study suggest that FGF/FgfR signaling enhances the intrinsic osteogenic potential by selectively expanding committed osteogenic cell populations as well as inversely regulating BMP-2 and noggin gene expression. PMID- 15780952 TI - IL-1-induced receptor activator of NF-kappa B ligand in human periodontal ligament cells involves ERK-dependent PGE2 production. AB - Periodontitis, an inflammatory disorder of the supporting tissue of teeth, is one of the most common infectious diseases in humans. Periodontal pathogens promote inflammatory cytokines such as interleukin-1 (IL-1) and prostaglandin E2 (PGE2), resulting in alveolar bone destruction. In the present study, we examined the cellular and molecular mechanisms of IL-1-induced osteoclastogenesis using a coculture system of human periodontal ligament (PDL) cells and mouse spleen cells. IL-1alpha induced tartrate-resistant acid phosphatase positive (TRAP+) cell formation in a dose-dependent manner. IL-1alpha up-regulated receptor activator of NF-kappaB ligand (RANKL) and down-regulated osteoprotegerin (OPG) mRNA expression in PDL cells. The addition of cell-permeable PKI, an inhibitor of the cAMP/PKA signaling pathway, to the cocultures 8 h after the IL-1alpha stimulation inhibited IL-1alpha-induced TRAP+ cell formation. IL-1alpha-induced TRAP+ cell formation was completely blocked by either NS398, a selective inhibitor of cyclooxygenase (COX)-2, or PD98059, a specific inhibitor of extracellular signal-regulated kinase (ERK). Pretreatment with NS398 and PD98059 also inhibited both the up-regulation of RANKL and the down-regulation of OPG expression by IL-1alpha in PDL cells. IL-1alpha activated ERK phosphorylation and PD98059 greatly inhibited both COX-2 mRNA expression and PGE(2) production induced by IL-1alpha in PDL cells. In contrast, NEMO binding domain (NBD) peptide, a specific inhibitor of NF-kappaB signaling, did not affect COX2, RANKL, or OPG mRNA expression induced by IL-1alpha. These results suggest that IL-1alpha stimulates osteoclast formation by increasing the expression level of RANKL versus OPG via ERK-dependent PGE2 production in PDL cells. PMID- 15780953 TI - Regulation by ultrasound treatment on the integrin expression and differentiation of osteoblasts. AB - It has been shown that ultrasound (US) stimulation accelerates fracture healing in the animal models and in clinical studies. However, the mechanism by which US achieves these outcomes is not clear. Here we investigated the effect of US stimulation on the differentiation of osteoblasts and osteoclastogenesis. The effect of different intensities of US stimulation (1 MHz, continuous wave) on the osteoblastic cell line MC3T3-E1 or primary cultured osteoblasts was examined. Flow cytometry showed that US stimulation at 125 mW/cm2 for 10 min transiently increased the surface expression of alpha2, alpha5, and beta1 integrins in both MC3T3-E1 and primary osteoblasts. Fluorocytochemistry showed that the actin cytoskeleton also reorganized in response to US stimulation. When the MC3T3-E1 cells were cultured in differentiation medium containing vitamin C and beta glycerophosphate, long-term US stimulation (10 min/day for 11 days) increased mineralized nodule formation, collagen content, and alkaline phosphatase activity. The intensity at 125 mW/cm2 exerts the most prominent action. Effect of long-term US stimulation on the osteoclastogenesis was also examined. US stimulation at a power of 62.5 or 125 mW/cm2 markedly inhibited RANKL plus M-CSF induced osteoclastic differentiation from bone marrow stromal cells. These findings suggest that US has a regulatory effect on the integrin expression and the differentiation of osteoblasts and osteoclastogenesis, which may contribute to the beneficial effects of US on the fracture repair. PMID- 15780954 TI - Suramin interacts with RANK and inhibits RANKL-induced osteoclast differentiation. AB - Suramin is a naphthalene trisulfonic acid derivative that inhibits osteoclast differentiation and bone resorption in vitro and in vivo; however, the mechanisms underlying this activity have not been studied. Receptor activator of NF-kB (RANK) ligand (RANKL) is a key regulator of osteoclast differentiation and function and this study evaluated the ability of suramin, which has been shown to disrupt protein-protein interactions, to interfere with RANKL functional activity and binding to RANK. Suramin inhibited osteoclastic bone resorption in a calvarial model and inhibited osteoclast differentiation in RANKL-stimulated murine spleen cells and RAW264.7 cells. RANKL-induced second messenger signaling (AKT and p38 MAP Kinase phosphorylation) was completely blocked by 100 microM suramin. The ability of RANKL to bind to recombinant human RANK-Fc (rhRANK-Fc) was reduced 50% by suramin in an in vitro binding assay. Surface plasmon resonance technology and nuclear magnetic resonance (NMR) were used to evaluate the ability of suramin to bind to rhRANK-Fc. Suramin was found to selectively interact with immobilized rhRANK-Fc chimera in a concentration-dependent manner by Biacore 3000 analysis. Similar results were obtained using saturation transfer difference NMR spectroscopy to demonstrate that suramin binds to rhRANK-Fc, but not IgG1Fc or sRANKL. In summary, these findings demonstrate that suramin inhibits sRANKL-induced osteoclast differentiation and suggest that these effects are mediated by suramin binding to RANK and blocking the ability of sRANKL to induce second messenger signaling. PMID- 15780955 TI - Efficacy of bioabsorbable antibiotic containing bone screw in the prevention of biomaterial-related infection due to Staphylococcus aureus. AB - Impregnation of antimicrobial agents within biodegradable orthopedic implants provides a possibility for local antimicrobial prophylaxis of biomaterial-related infections. The objective of this study was to evaluate the efficacy of a bioabsorbable ciprofloxacin containing bone screw (Ab-PLGA) in the prevention of biomaterial-related infection due to Staphylococcus aureus in a rabbit model. Animals in Group I (n=8) received a Ab-PLGA screw contaminated with S. aureus, while animals in Group II (n=8) received a stainless steel (SS) screw contaminated with S. aureus. In two negative control groups, the animals received a Ab-PLGA screw (Group III, n=4) or a SS screw (Group IV, n=4) without bacterial contamination. 18F-FDG-PET imaging, performed at 6 weeks, was applied as a novel quantitative in vivo imaging modality of implant-related infection. Infection was verified by swab cultures, direct cultures of the retrieved implant, and quantitative cultures of pulverized bone. The concentrations of ciprofloxacin in serum and local bone tissue were determined by a high performance liquid chromatographic (HPLC) method with fluorescence (FLD) detection. In the group of contaminated Ab-PLGA screws, all cultures were negative. In the group of contaminated SS screws, all cultures of retrieved implants and six cultures out of eight of pulverized bone were positive for inoculated S. aureus. In negative control groups, all cultures were negative except one contaminant (S. cohnii) found in a SS screw culture. Verified infection of contaminated SS screws was collaborated by the increased 18F-FDG-PET uptake (P=0.004 compared with the group of contaminated Ab-PLGA screws). The mean bone tissue concentration of ciprofloxacin varied from 2.54 to 0.83 microg/g bone as a function of distance from the implantation site. The serum concentration of ciprofloxacin remained undetectable and below the resolution of the analytic method (<5.0 ng/ml). This study confirmed the in vivo efficacy of bioabsorbable antibiotic containing bone screw in the prevention of biomaterial-related infection due to S. aureus. PMID- 15780956 TI - Tumor necrosis factor alpha (TNF-alpha) coordinately regulates the expression of specific matrix metalloproteinases (MMPS) and angiogenic factors during fracture healing. AB - Recent studies from our laboratory demonstrate that TNF-alpha signaling contributes to the regulation of chondrocyte apoptosis and a lack of TNF-alpha signaling leads to a persistence of cartilaginous callus and delayed resorption of mineralized cartilage. This study examines how delays in the endochondral repair process affect the expression of specific mediators of proteolytic cartilage turnover and vascularization. Simple closed fractures were produced in wild type and TNF-alpha receptor (p55-/-/p75-/-)-deficient mice. Using ribonuclease protection assay (RPA) and microarray analysis, the expression of multiple mRNAs for various angiogenic factors and the metalloproteinase gene family were measured in fracture calluses. The direct actions of TNFalpha on the expression of specific angiogenic factors and metalloproteinases (MMPs) was examined in both cultured callus cells and articular chondrocytes to compare the effects of TNF-alpha in growth cartilage versus articular cartilage. MMPs 2, 9, 13, and 14 were quantitatively the most prevalent metalloproteases and all showed peaks in expression during the chondrogenic period. In the absence of TNF-alpha signaling, the expression of all of these mRNAs was reduced. The angiopoietin families of vascular regulators and their receptors were expressed at much higher levels than the VEGFs and their receptors and while the angiopoietins showed diminished or delayed expression in the absence of TNF-alpha signaling, VEGF and its receptors remained unaltered. The expression of vascular endothelial growth inhibitor (VEGI or TNFSF15) showed a near absence in its expression in the TNF alpha receptor-deficient mice. In vitro assessment of cultured fracture callus cells in comparison to primary articular chondrocytes showed that TNF-alpha treatment specifically induced the expression of MMP9, MMP14, VEGI, and Angiopoietin 2. These results suggest that TNF-alpha signaling in chondrocytes controls vascularization of cartilage through the regulation of angiopoietin and VEGI factors which play counterbalancing roles in the induction of growth arrest, or apoptosis in endothelial cells. Furthermore, TNF-alpha appears to regulate, in part, the expression of two key proteolytic enzymes, MMP 9 and MMP14 that are known to be crucial to the progression of vascularization and turnover of mineralized cartilage. Thus, TNF-alpha signaling in healing fractures appears to coordinate the expression of specific regulators of endothelial cell survival and metalloproteolytic enzymes and is essential in the transition and progression of the endochondral phase of fracture repair. PMID- 15780957 TI - Human femoral neck has less cellular periosteum, and more mineralized periosteum, than femoral diaphyseal bone. AB - Periosteal expansion enhances bone strength and is controlled by osteogenic cells of the periosteum. The extent of cellular periosteum at the human femoral neck, a clinically relevant site, is unclear. This study was designed to histologically evaluate the human femoral neck periosteal surface. Femoral neck samples from 11 male and female cadavers (ages 34-88) were histologically assessed and four periosteal surface classifications (cellular periosteum, mineralizing periosteum, cartilage, and mineralizing cartilage) were quantified. Femoral mid-diaphysis samples from the same cadavers were used as within-specimen controls. The femoral neck surface had significantly less (P<0.05) cellular periosteum (18.4+/-9.7%) compared to the femoral diaphysis (59.2+/-13.8%). A significant amount of the femoral neck surface was covered by mineralizing periosteal tissue (20-70%). These data may provide an alternate explanation for the apparent femoral neck periosteal expansion with age and suggest the efficiency of interventions that stimulate periosteal expansion may be reduced, albeit still possible, at the femoral neck of humans. PMID- 15780958 TI - An effective case of growth hormone treatment on cartilage-hair hypoplasia. AB - Cartilage-hair hypoplasia (CHH) is an autosomal recessive metaphyseal chondrodysplasia characterized by severe short-limb short stature and hypoplastic hair. The responsible gene for CHH has been identified to be ribonuclease of mitochondrial RNA-processing (RMRP) gene. We examined RMRP genes of a 3-year-old Japanese CHH boy and his family and revealed a novel mutation: 20 bp duplication (TACTCTGTGAAGCTGAGGAC), in promoter region of maternal allele, at nucleotide -3 and a reported 218A>G point mutation in transcribed region of paternal allele. No treatment for CHH has been established so far. Growth hormone (GH) action has its effect on linear growth and on bone remodeling and homeostasis. Recently, GH has been used to improve severe short stature caused by not only GH deficiency (GHD) but also some skeletal dysplasias including achondroplasia. To improve severe short stature, we treated the patient with 0.175 mg kg-1 week-1 of GH for 7 years. His height was improved from -4.2 SD to -3.0 SD by 1 year of GH treatment. Following treatment had given positive effects continuously on his height to -2.6 SD by 3.1 years GH medication. Then, when he was 6 years old, surgical lengthening was performed and his height reached to -2.0 SD. After the surgery, we continued GH treatment. Additional GH treatment of 3.6 more years had kept his height to -2.0 SD. However, when he was 8 years old, because there was an interruption of GH treatment, the velocity of his height was obviously decreased comparing before and during the interruption, which was calculated 3.4 and 2.2 cm/year, respectively, and the SD score was decreased to -2.1 SD. This result of total 7 years of GH treatment suggested that GH treatment significantly improved his disturbed bone growth and had also positive efficacy to keep growth rate. This result implies the connection between GH signal and RMRP gene. Additionally, GH may be considered to be an efficient treatment for CHH. PMID- 15780959 TI - Calcium phosphate scaffold and bone marrow for bone reconstruction in irradiated area: a dog study. AB - Squamous cell carcinomas (SCC) of the upper aero-digestive tract are characterized by a high incidence of bone invasion; their treatment often requires large and damaging surgical resections and radiotherapy. Surgical and radiotherapeutic procedures generate irreversible effects on normal tissues, involving injuries on their reparation properties, especially on bone. The quality of life of patients undergoing major surgery and radiotherapy in maxillary and mandible areas is often reduced but could be improved by bone reconstructions. Bone reconstructions are rarely performed because surgery is complex and unsafe in irradiated bone. The aim of the study was to evaluate the bone reconstruction possibilities of macroporous biphasic calcium phosphate (MBCP) associated to autologous bone marrow (BM) graft injected after irradiation. MBCP hollowed blocks were specially designed and implanted in tibia and femur bone before irradiation in a dog model. Implants were removed after 18 weeks. This is the first report of experiments performed after radiation delivery using high fractionated doses approximating usual treatment of SCC in human. The quality of the bone adjacent to implanted MBCP and the bone ingrowth's rates were evaluated. The qualitative and quantitative role of BM grafts associated with the MBCP implants was determined, using scanning electron microscopy linked to quantitative image analysis. A direct contact between newly formed bone and MBCP implants associated to BM graft was observed, without fibrous interposition. The new-bone formation was statistically increased inside the MBCP (P=0.0126) by BM grafts. This study demonstrates that BM graft added to MBCP constitute an appropriate material to be considered in case of bone defect occurring in irradiated tissue, and could be foreseen for use after bone removal for oncologic obligations. PMID- 15780960 TI - Deteriorated geometric structure and strength of the midfemur in men with complete spinal cord injury. AB - Spinal cord injury (SCI) results in a dramatic loss of bone mineral and a marked increase in fracture incidence in the femur; however, its effect on the femur's geometric structure and strength is poorly studied. The primary purpose of the present study was to assess the geometric structure, composition, and strength of the midfemur in men with long-term (>2 years), complete SCI (C6-L1 level; n=7) relative to men without SCI (n=8). T1-weighted axial images of the thigh were collected on a GE 1.5-T magnetic resonance imager and geometric, structure, composition, and strength measurements of the midfemur and skeletal muscle volume of the midthigh were determined. Areal bone mineral density (aBMD), bone mineral content (BMC), and bone area of the midthird of the femur and arms were determined using dual-energy X-ray absorptiometry. There were no differences in age, height, weight, femur length, arm BMC, arm aBMD, or arm bone area between the SCI group and controls. While the volume of the midfemur was not different in the two groups, the medullary cavity had 53% more volume and was 21-25% wider in the SCI group (P<0.05). In contrast, the cortical wall in the SCI group had a 24% lower volume and was 27-47% thinner (P<0.05). The cortical wall was particularly thin in the posterior section of the bone. The SCI group also had lower BMC and aBMD in the midfemur (21% and 25%, respectively, P<0.05). Calculated cross sectional moment of inertia (CSMI), section modulus (Z), and polar moment of inertia (J) were lower in the SCI group (13-19%, P<0.05). A higher ratio of cortical bone volume to muscle volume and BMC to muscle volume in the SCI group (P<0.05) suggests that there was a greater loss of muscle than cortical bone after SCI; however, muscle volume was strongly correlated with cortical bone volume and BMC in the SCI and control groups (r=0.71 to 0.90, P<0.05). Muscle volume was also moderately to strongly correlated with CSMI and Z in the anterior posterior direction and J. Muscle volume was weakly correlated or not correlated with bone strength measures in the control group (P>0.05). These findings suggest that after SCI, the midfemur erodes on the endosteal surface, resulting in a decreased resistance to bending and torsion. Although midthigh muscle volume appears to decline to a greater degree than midfemur cortical bone volume and BMC, their relationships remain strong. PMID- 15780961 TI - Relationship between compressive properties of human os calcis cancellous bone and microarchitecture assessed from 2D and 3D synchrotron microtomography. AB - The aim of this study was to determine the contribution of 2D and 3D microarchitectural characteristics in the assessment of the mechanical strength of os calcis cancellous bone. A sample of cancellous bone was removed in a medio lateral direction from the posterior body of calcaneus, taken at autopsy in 17 subjects aged 61-91 years. The sample was first used for the assessment of morphological parameters from 2D morphometry and 3D synchrotron microtomography (microCT) (spatial resolution=10 microm). The 2D morphometry was obtained from three slices extracted from the 3D microCT images. Very good concordance was shown between 3D microCT slices and the corresponding physical histologic slices. In 2D, the standard histomorphometric parameters, fractal dimension, mean intercept length, and connectivity were computed. In 3D, histomorphometric parameters were computed using both the 3D mean intercept length method and model independent techniques. The 3D fractal dimension and the 3D connectivity, assessed by Euler density, were also evaluated. The cubic samples were subjected to elastic compressive tests in three orthogonal directions (X, Y, Z) close to the main natural trabecular network directions. A test was performed until collapse of trabecular network in the main direction (Z). The mechanical properties were significantly correlated to most morphological parameters resulting from 2D and 3D analysis. In 2D, the correlation between the mechanical strength and bone volume/tissue volume was not significantly improved by adding structural parameters or connectivity parameter (nodes number/tissue volume). In 3D, one architectural parameter (the trabecular thickness, Tb.Th) permitted to improve the estimation of the compressive strength from the bone volume/tissue volume alone. However, this improvement was minor since the correlation with the BV/TV alone was high (r=0.96). In conclusion, which is in agreement with the statistic's rules, we found, in this study, that the determination of the os calcis bone compressive strength using the 3D bone volume fraction cannot be improved by adding 3D architectural parameters. PMID- 15780962 TI - Skeletal age deviation assessed by the Tanner-Whitehouse 2 method is associated with bone mass and fracture risk in children. AB - The aim of this population-based case-control study was to describe the association among skeletal age deviation (SAD), bone density, and upper limb fracture risk in male and female children aged 9-16 years. A total of 321 fracture cases and 321 randomly selected individually matched controls were studied. Skeletal age was assessed by standard atlas. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DXA) and metacarpal index (MI). There were no significant differences in mean skeletal age or chronological age between fracture cases and controls. However, SAD was associated with total, hand, and female fracture risk (all P<0.05). The fracture associations became nonsignificant after adjustment for BMD and MI in all subgroups with the exception of hand fractures (OR, 0.67/year; 95% CI, 0.47-0.96). SAD was also positively associated with BMD at all sites (r=0.33-0.35, all P<0.05) and MI (r=0.20, P<0.05). The strength of association reduced but remained significant at most sites after adjustment for body size, maturity, age, and sex. In conclusion, SAD is positively associated with measures of bone strength and negatively associated with upper limb fracture risk (especially those of the hand) in children. SAD is simple to measure and gives additional information regarding bone health and fracture risk in children. PMID- 15780963 TI - The efficacy and tolerability of risedronate on bone mineral density and bone turnover markers in osteoporotic Chinese women: a randomized placebo-controlled study. AB - Osteoporosis has become an important health problem in postmenopausal Asian populations as the prevalence of hip and vertebral fractures in some Asian countries has risen to approach that of Caucasian populations. Risedronate, a pyridinyl-bisphosphonate agent, is a potent inhibitor of bone resorption. Risedronate increases bone mineral density (BMD), reduces markers of bone turnover, and reduces the risk of fractures in Caucasian postmenopausal women. To determine the efficacy and tolerability of risedronate in Chinese, a multicenter, randomized, double blind, placebo controlled study was performed in Hong Kong. Sixty-five (65) postmenopausal osteoporotic Southern Chinese women, aged 67+/-6 years, were randomly assigned to receive either risedronate 5 mg daily (n=31) or placebo (n=34) for 12 months. All women received calcium carbonate 500 mg daily and vitamin D 400 IU daily. Mean baseline BMD T-score at the spine and total hip was -3.4 and -2.6, respectively. A significant increase in spine BMD was already evident at month 3 of risedronate treatment (P<0.001). Risedronate significantly increased BMD and reduced bone turnover markers as compared with placebo. The risedronate group had significant increase in BMD at 12 months at both the spine and hip when compared with the placebo group (L1-4 6.6% vs. 0.4%, P<0.001; total hip 2.7% vs. 0.3, P<0.0001; femoral neck 1.8% vs. 1.1%, P<0.02; trochanter 4% vs. 1.1%, P<0.0001, respectively). Significant changes in urine N-telopeptide (NTx) and serum osteocalcin were evident as early as 1 and 3 months, respectively, with risedronate treatment. No significant changes were seen in both BMD and bone markers in the placebo group. Risedronate was well tolerated without major adverse effects. We conclude that risedronate is an effective and well-tolerated agent for the treatment of postmenopausal osteoporosis in Asian population. PMID- 15780964 TI - Association of ANKH gene polymorphisms with radiographic hand bone size and geometry in a Chuvasha population. AB - We performed a family-based association study to test the hypothesis that genetic variation at the human orthologue of the mouse progressive ankylosis gene (ANKH) is involved in determining bone size (BS) and bone geometry (BG). The study population comprised 126 nuclear families with 574 adult Chuvashian individuals living in small villages in the Russian Federation. Quantitative bone traits were determined by analyzing plain hand radiographs. Familial correlations for all studied traits revealed a high degree of heritability in this ethnically homogeneous population. Three simple tandem repeat (STR) polymorphisms, one intragenic and two flanking markers, as well as six single nucleotide polymorphisms (SNPs) were tested. The SNPs were detected by re-sequencing experiments and covered ANKH exons with their flanking splice sites and the promoter region. We used three different transmission disequilibrium tests (TDTs) and obtained multiple significant association signals for all investigated bone traits. Alleles of several markers located at different positions of the ANKH locus, including the promoter, consistently revealed the association. The bone traits tested are closely related to bone fragility suggesting a role for ANKH in osteoporosis. PMID- 15780965 TI - Bone structure and volumetric density in young adults born prematurely: a peripheral quantitative computed tomography study. AB - Prematurity is known to be associated with reduced bone mineral density (BMD) in childhood, but whether this condition has long-term detrimental consequences on adult bone structure is not known. In this study, we measured with peripheral quantitative computed tomography (pQCT) the total bone cross-sectional area, cortical area and wall thickness, cortical and trabecular density, and a density weighed polar section modulus as a bone strength index (BSI) at distal and shaft sites of right radius and tibia in a group of 40 prematurely born, otherwise normally developed and healthy young adults (17 women and 23 men, aged 18 to 27 years) and compared their data to corresponding data obtained from a group of 42 control subjects born term (20 women and 22 men, aged 18 to 28 years). Body height and weight were similar in both groups, but the preterm group had significantly lower BSI values at distal sites of tibia (approximately -16%) and radius (approximately -13%) and at tibial shaft (approximately -11%) as compared to control group. In the weight-bearing tibia, BMC was lower and the lower BSI values were mainly due to smaller total bone cross-sectional area. For unknown reason, this prematurity-associated detrimental effect seemed to concern more men than women. In contrast, prematurity was not associated with volumetric trabecular and cortical densities at any measured bone site while the typical sex differences in bone density were observed. We conclude that prematurity is associated with somewhat smaller cross-sectional bone dimensions in terms of body size in young adulthood. Due to the cross-sectional design, this study could not reveal specific reasons but they may pertain to nutrition during the neonatal period and living habits in general. PMID- 15780966 TI - Different susceptibility of human immunodeficiency virus type 1 to Env gp41 derived synthetic peptides corresponding to the C-terminal heptad repeat region. AB - Two functional domains, alpha-helical heptad repeat 1 (HR-1) and HR-2, located in the N-terminal and C-terminal regions of human immunodeficiency virus type 1 (HIV 1) Env gp41, respectively, play an important role in the fusion process. Synthetic 34-amino-acid peptide that contains the HR-2 region, named C34, has been shown to inhibit the HIV-1 fusion process. Here, we prepared six representative peptides (C34-B1, -B2, -A, -C1, -C2, and -E from subtypes B, A, C, and E, respectively) according to the sequences from the HIV sequence database of Los Alamos. All the C34 peptides had lower ability to inhibit the primary isolates (subtypes B and CRF01_AE) than subtype B laboratory strain LAI. On the other hand, the L-2 cell clone, isolated from persistently LAI-infected MT-4 cells (MT-4/LAI), showed unique C34 peptide sensitivities. L-2 virus has the same sequences at HR-1 and HR-2 regions as LAI, but showed higher syncytia formation activity than LAI. Interestingly, the sensitivity of L-2 was higher to C34-B2 and -A but slightly lower to C34-C1 at higher concentrations than MT-4/LAI, while C34 B1, -C2, and -E showed similar activity against both viruses. Thus, in addition to the sequences of the C34 peptide as well as of the HR-1 and HR-2 regions in target viruses used for fusion assays, the fusion inhibitory activities of C34 peptides seem to be affected by viral factor(s) other than the gp41 alpha-helical heptad repeats. PMID- 15780967 TI - Human genome-wide screen of haplotype-like blocks of reduced diversity. AB - An important proportion of the human genome is organized in regions of high linkage disequilibrium (LD) and low haplotype diversity, referred to as haplotype blocks. Here, we perform a genome-wide screen of haplotype-like blocks presenting just two main haplotypes at a frequency higher than 1%, based on single nucleotide polymorphism (SNP) frequencies from two populations: African-Americans and Caucasians, using data from the Celera SNP database. These haplotype-like blocks of reduced diversity are more abundant and of longer size in Caucasians, in agreement with population history. Several of the discovered blocks are good candidates for targets of natural selection, such as those blocks containing a cluster of bitter taste receptors or the apolipoprotein L1. In addition, several genes putatively involved in susceptibility to common diseases are included in these haplotype-like blocks of reduced diversity. This fact may present important implications in association studies, leading to a reduction of genotyping efforts. PMID- 15780968 TI - Intraspecific variation in fem-3 and tra-2, two rapidly coevolving nematode sex determining genes. AB - The sex determination gene fem-3 encodes one of the most divergent proteins yet described in the terrestrial nematode Caenorhabditis. Despite this rapid sequence change, however, FEM-3 is essential for male development in the three species surveyed thus far. It also participates in conserved protein-protein complexes with the transmembrane receptor TRA-2 and the phosphatase FEM-2 in these species. These interactions show strong species specificity, indicating that conserved residues are not sufficient for function and that compensatory evolution between binding partners is important. To shed further light on the nature of this coevolution, and to discern the extent of amino acid polymorphism allowed in FEM 3 and the domain of TRA-2 that binds it, we have examined intraspecific variation in the gonochoristic species Caenorhabditis remanei. Ten new complete Cr-fem-3 alleles from three regions of the United States are described. We also obtained sequences for the FEM-3-binding domain of TRA-2 for 9 of the same strains. These alleles were compared with each other, with the European founder alleles, and with the orthologous sequences from the congeners Caenorhabditis elegans and C. briggsae. We find that FEM-3 harbors abundant amino acid polymorphisms along its entire length. The majority (but not all) of these occur in nonconserved residues, and in at least one domain there is evidence for diversifying selection. The FEM-3-binding domain of TRA-2 is less polymorphic than FEM-3. Amino acids neither polymorphic nor conserved between species are candidates for residues mediating species-specific interaction of FEM-3 with its binding partners. PMID- 15780969 TI - Analysis of endogenous S1P and LPA receptor expression in CHO-K1 cells. AB - The CHO-K1 cell line is commonly used for studies of recombinantly expressed proteins, including proteins of the G protein-coupled receptor (GPCR) family. This laboratory has used CHO-K1 cells for the functional characterization of Edg family GPCRs. However, parental CHO-K1 cells respond to lysophospholipids in in vitro functional assays, which suggests expression of endogenous Edg family GPCRs. To determine the repertoire of Edg family receptor expression in this cell line, alignments of human and rodent sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) receptor sequences were used to design semi-redundant oligonucleotide pairs. A portion of each receptor gene coding sequence was amplified from Chinese hamster genomic DNA and the resultant gene fragments sequenced. Species-specific oligonucleotide pairs were designed using this novel sequence information and used to detect expression of S1P(1,2,4) and LPA(1) transcripts in CHO-K1 cells by RT-PCR. PMID- 15780970 TI - Selenium binding to human hemoglobin via selenotrisulfide. AB - Selenotrisulfide (e.g., glutathione selenotrisulfide (GSSeSG)) is an important intermediate in the metabolism of selenite. However, its reactivity with biological substances such as peptides and proteins in the subsequent metabolism is still far from clearly understood, because of its chemical instability under physiological conditions. Penicillamine (Pen) is capable of generating a chemically stable and isolatable selenotrisulfide, PenSSeSPen. To explore the metabolic fate of selenite in red blood cells (RBC), we investigated the reaction of selenotrisulfide with human hemoglobin (Hb) using PenSSeSPen as a model. PenSSeSPen rapidly reacted with Hb under physiological conditions. From the analysis of selenium binding using the Langmuir type binding equation, the apparent binding number of selenium per Hb tetramer almost corresponded to the number of reactive thiol groups of Hb. The thiol group blockade of Hb by iodoacetamide treatment completely inhibited the reaction of PenSSeSPen with Hb. In addition, MALDI-TOF mass spectrometric analysis of the selenium-bound Hb revealed that PenSSe moiety binds to the beta subunits of Hb. Overall, the reaction of PenSSeSPen with Hb appears to involve the thiol exchange between Pen and the cysteine residues on the beta subunit of Hb. PMID- 15780971 TI - "In vitro" protection of DNA from Fenton reaction by plant polyphenol verbascoside. AB - The protection effect of verbascoside (Ver) against Fenton reaction on plasmid pBR322 DNA was studied using agarose gel electrophoresis and UV-visible spectroscopy. The pBR322 plasmid DNA is damaged by hydroxyl radical (OH*) generated from the Fenton reaction with H2O2 and Fe(II) or Fe(III). This DNA damage is characterized by the diminution of supercoiled DNA forms or by the increase of relaxed or linear DNA forms after oxidative attack. The UV spectrum study showed that verbascoside can form complexes with Fe(II) or Fe(III), and the complexation can be reversed by the addition of EDTA. The formation constants of verbascoside-Fe complexes were estimated as 10(21.03) and 10(31.94) M(-2) for Fe(II) and Fe(III) respectively. The inhibition of Fenton reaction by verbascoside could be partially explained by the sequestration of Fe ions. PMID- 15780972 TI - Two novel arginine/serine (SR) proteins in maize are differentially spliced and utilize non-canonical splice sites. AB - The serine-arginine (SR)-rich splicing proteins are highly conserved RNA binding nuclear phosphor-proteins that play important roles in both regular and alternative splicing. Here we describe two novel putative SR genes from maize, designated zmRSp31A and zmRSp31B. Both genes contain characteristic RNA binding motifs RNP-1 and RNP-2, a serine/arginine-rich (RS) domain and share significant sequence similarity to the Arabidopsis atRSp31 family of SR proteins. Both zmRSp31A and zmRSp31B produce multiple transcripts by alternative splicing, of which majority of the alternatively spliced transcripts utilize non-canonical splice sites. zmRSp31A and zmRSp31B produce at least six and four transcripts, respectively, of which only one corresponds to the wild type proteins for each gene. All the alternatively spliced transcripts of both the genes, with one exception, are predicted to encode small truncated proteins containing only the RNP-2 domain of their first RNA recognition motif and completely lack the carboxyl terminal RS domain. We provide evidence that some of the alternatively spliced transcripts of both genes are associated with polysomes and interact with the translational machinery. PMID- 15780973 TI - No major effect of the insulin-like growth factor I gene on bone mineral density in premenopausal Chinese women. AB - Osteoporosis is a major public health problem, mainly characterized by low bone mineral density (BMD). BMD is a complex trait that is determined by multiple genes. Insulin-like growth factor I (IGF-I) is an important growth factor of bone and thus IGF-I gene has been considered as an attractive candidate gene for osteoporosis. A few studies on the relationship between variants of the IGF-I gene and BMD variation, via traditional association and/or linkage methods, have yielded conflicting results. In this study, we simultaneously tested association and/or linkage of a cytosine-adenine (CA) repeat polymorphism at 1 kb upstream of the transcription initiation site of the IGF-I gene with BMD variation in a large cohort of premenopausal Chinese women. A total of 1263 subjects from 402 Chinese nuclear families were examined. Each family consists of both parents and at least one daughter aged between 20 and 45 years. BMDs (g/cm(2)) at the lumbar spine and hip were measured using dual-energy X-ray absorptiometry (DXA). Applying the QTDT (quantitative transmission disequilibrium tests) progam, we did not find significant evidence of association or linkage between the CA repeat polymorphism of the IGF-I gene and BMD variation at any skeletal site. Our data do not support the IGF-I gene having major effect on BMD variation in premenopausal Chinese women. PMID- 15780974 TI - Twisted gastrulation and chordin inhibit differentiation and mineralization in MC3T3-E1 osteoblast-like cells. AB - Bone morphogenetic proteins (BMPs) are potent inducers of osteoblast differentiation. The accessibility of BMP ligands for binding to their receptors is regulated by secreted proteins Twisted gastrulation (Tsg) and Chordin (Chd). Tsg antagonizes BMP signaling by forming ternary complexes with Chd and BMPs, thereby preventing BMPs from binding to their receptors. In addition to the anti BMP function, Tsg also has pro-BMP activity, partly mediated by cleavage and degradation of Chd, which releases BMPs from ternary complexes. The roles of Tsg and Chd in osteoblast differentiation are not known. Therefore, in the present study, we investigated the effect of exogenous Tsg and Chd on osteoblast differentiation and mineralization using a well-characterized subclone of MC3T3 E1 osteoblast-like cells. Our results show that Tsg and Chd are expressed in MC3T3-E1 osteoblast-like cells. While Tsg mRNA levels decrease during osteoblast differentiation, Chd levels are found to increase. Tsg and Chd proteins accumulate in the cell culture media as the osteoblasts differentiate. Exogenous Tsg and Chd inhibit osteoblast differentiation and mineralization. Osteocalcin (OCN) mRNA levels decrease following both Tsg and Chd treatment. Tsg and Chd also inhibit alkaline phosphatase (ALP) activity in a dose-dependent manner. To provide insight into the mechanism of Tsg and Chd action, we investigated the effect of Tsg and Chd on BMP activity by determining phosphorylated Smad1 (pSmad1) levels. We show that both Tsg and Chd can independently and in combination reduce pSmad1 levels in MC3T3-E1 cells treated with BMP4. Further, BMP2 partially reverses the inhibitory effect of Tsg and Chd on ALP activity. Taken together, these results suggest that Tsg and Chd are involved in osteoblast differentiation and mineralization by regulating BMP signaling. PMID- 15780975 TI - Parathyroid hormone and rates of bone formation are raised in perimenopausal rural Gambian women. AB - To investigate rates of bone turnover and calcium homeostasis in Gambian women, we recruited 103 peri- and postmenopausal women, aged 45 to 80+ years and 11 women of reproductive age. Fasting blood was analyzed for plasma osteocalcin, PTH, 25-hydroxyvitamin D [25(OH)D], 1,25-dihydroxyvitamin D [1,25(OH)(2)D], total and bone-specific alkaline phosphatase. Plasma and urinary calcium, inorganic phosphate, sodium, potassium, creatinine, and albumin and urine free deoxypyridinoline (Dpd) was also measured. Samples from 20 premenopausal and 31 postmenopausal women from Cambridge, UK were analyzed, using the same methodology for comparison. For the Gambian women, peak calcium excretion occurred at around 50 years of age. For women aged > or =45 years, calcium excretion decreased by 3.0% per year of age (SE 1%; P < 0.005). In this age group, 25(OH)D also decreased with age (P < 0.005). Urinary sodium output, pH, and titratable acid output decreased (all P < 0.05) and total alkaline phosphatase (P < 0.005), osteocalcin (P < 0.005), and PTH (P < 0.05) increased with age. Comparisons were made between the following groups of Gambian and British women: premenopausal, early (age 55-64 years)- and late (age 65+ years)-postmenopausal. Gambian women of all ages were lighter (P < 0.001), shorter (P < 0.01), and had higher plasma bone-specific alkaline phosphatase activity (P < 0.05) and higher concentrations of osteocalcin (P < 0.05), PTH (P < 0.001), 1,25(OH)(2)D (P < 0.001), and 25(OH)D (P < 0.001). There were no consistent differences in calcitonin, while urinary free Dpd outputs were lower in the Gambians (P < 0.001). Plasma calcium, phosphate, and albumin (P < 0.01) were significantly lower. Urinary calcium, phosphate, sodium, and potassium excretion were lower, particularly in the postmenopausal group (P < 0.001). Although Gambian urine pH was more acidic, titratable acid output was lower (P < 0.01). These data show that Gambian women with low dietary calcium intakes and good vitamin D status have low urinary calcium excretion and that menopausal changes in calcium and bone metabolism among Gambian women are similar to those seen in other populations. In addition, they demonstrate that Gambian women of all ages have raised plasma PTH and 1,25(OH)(2)D concentrations and raised markers of osteoblast activity. We postulate that high endogenous PTH concentrations may be beneficial to bone health in Gambian women, removing fatigue damage and improving bone quality. PMID- 15780976 TI - Increased adipogenesis in bone marrow but decreased bone mineral density in mice devoid of thyroid hormone receptors. AB - Mice deficient for all known thyroid hormone receptors, TRalpha1-/-beta-/- mice, display a clear skeletal phenotype characterized by growth retardation, delayed maturation of long bones and decreased trabecular and total bone mineral density (BMD; -14.6 +/- 2.8%, -14.4 +/- 1.5%). The aim of the present study was to investigate the molecular mechanisms behind the skeletal phenotype in TRalpha1-/ beta-/- mice. Global gene expression analysis was performed on total vertebrae from wild-type (WT) and TRalpha1-/-beta-/- mice using DNA microarray and the results were verified by real-time PCR. The mRNA levels of six genes (AdipoQ, Adipsin, Fat-Specific Protein 27 (FSP 27), lipoprotein lipase (LPL), retinol binding protein (RBP) and phosphoenolpyruvate carboxykinase (PEPCK)) expressed by mature adipocytes were increased in TRalpha1-/-beta-/- compared with WT mice. An increased amount of fat (225% over WT) due to an increased number but unchanged mean size of adipocytes in the bone marrow of TRalpha1-/-beta-/- mice was revealed. Interestingly, the mRNA levels of the key regulator of osteoclastogenesis, receptor activator of NF-varkappab ligand (RANKL), were dramatically decreased in TRalpha1-/-beta-/- mice. In conclusion, TRalpha1-/-beta /- mice demonstrated increased expression of adipocyte specific genes and an increased amount of bone marrow fat. Thus, these mice have increased adipogenesis in bone marrow associated with decreased trabecular bone mineral density (BMD). One may speculate that these effects either could be caused by an imbalance in the differentiation of the osteoblast and the adipocyte lineages at the expense of osteoblastogenesis, or by independent effects on the regulation of both osteoblastogenesis and adipogenesis. PMID- 15780977 TI - A plant polyphenol-rich extract restores the suppressed functions of phagocytes in influenza virus-infected mice. AB - Influenza infection was induced in white ICR mice by intranasal (i.n.) inoculation of the virus A/Aichi/2/68 (H3N2). The number, migration and phagocyte indices of alveolar and peritoneal macrophages (pMO) and of blood polymorphonuclear leukocytes (PMNs), as well as the inhibition of the PMN adherence in the presence of a specific antigen were followed for 9 days after infection. The effect of the i.n. application of a polyphenol-rich extract, designated as polyphenolic complex (PC), isolated from the medicinal plant Geranium sanguineum L., on the inspected immune parameters was studied in parallel with the virological parameters of the infection, e.g. rate of mortality, mean survival time (MST), infectious lung virus titre and consolidation of the lungs. It was found that the application of PC induced a continuous 2- to 2.5-fold rise in the number of both peritoneal and alveolar macrophages (aMO) in the infected and healthy controls. The migration of both peritoneal and aMO increased 1.5- to 2-fold in the group of infected PC-treated animals and four to fivefold in the control group, the maximum being on day 9. PC stimulated phagocyte activities of blood PMNs in both infected and healthy mice. The leukocyte adherence inhibition (LAI) index decreased in the infected and PC treated animals. The restoration of the suppressed functions of phagocytes in influenza virus-infected mice (VIM) was consistent with a prolongation of MST and reduction in mortality rate, infectious virus titre and lung consolidation. The immunoenhancing properties of PC apparently contribute to the overall protective effect of the plant preparation in the lethal murine experimental influenza A/Aichi infection. PMID- 15780978 TI - Transcriptional and phylogenetic analysis of five complete ambystomatid salamander mitochondrial genomes. AB - We report on a study that extended mitochondrial transcript information from a recent EST project to obtain complete mitochondrial genome sequence for 5 tiger salamander complex species (Ambystoma mexicanum, A. t. tigrinum, A. andersoni, A. californiense, and A. dumerilii). We describe, for the first time, aspects of mitochondrial transcription in a representative amphibian, and then use complete mitochondrial sequence data to examine salamander phylogeny at both deep and shallow levels of evolutionary divergence. The available mitochondrial ESTs for A. mexicanum (N=2481) and A. t. tigrinum (N=1205) provided 92% and 87% coverage of the mitochondrial genome, respectively. Complete mitochondrial sequences for all species were rapidly obtained by using long distance PCR and DNA sequencing. A number of genome structural characteristics (base pair length, base composition, gene number, gene boundaries, codon usage) were highly similar among all species and to other distantly related salamanders. Overall, mitochondrial transcription in Ambystoma approximated the pattern observed in other vertebrates. We inferred from the mapping of ESTs onto mtDNA that transcription occurs from both heavy and light strand promoters and continues around the entire length of the mtDNA, followed by post-transcriptional processing. However, the observation of many short transcripts corresponding to rRNA genes indicates that transcription may often terminate prematurely to bias transcription of rRNA genes; indeed an rRNA transcription termination signal sequence was observed immediately following the 16S rRNA gene. Phylogenetic analyses of salamander family relationships consistently grouped Ambystomatidae in a clade containing Cryptobranchidae and Hynobiidae, to the exclusion of Salamandridae. This robust result suggests a novel alternative hypothesis because previous studies have consistently identified Ambystomatidae and Salamandridae as closely related taxa. Phylogenetic analyses of tiger salamander complex species also produced robustly supported trees. The D-loop, used in previous molecular phylogenetic studies of the complex, was found to contain a relatively low level of variation and we identified mitochondrial regions with higher rates of molecular evolution that are more useful in resolving relationships among species. Our results show the benefit of using complete genome mitochondrial information in studies of recently and rapidly diverged taxa. PMID- 15780979 TI - Nested genes: biological implications and use of AFM for analysis. AB - A "nested" gene is located within the boundaries of a larger gene, often within an intron and in the opposite orientation. Such structures are common in bacteria and viruses, but have also been described in higher species as diverse as Drosophila and humans. Expression of nested and host genes may be simultaneously up-regulated due to use of common enhancers, or down-regulated through steric hindrance or interference caused by annealing of the complementary RNAs, leading to degradation. Methods for RNA analysis such as RT-PCR and in situ hybridization reveal the presence of specific mRNAs, but do not address regulation of expression within a single cell at a single genetic locus. Atomic force microscopy is a relatively new technology, which allows visualization of the movement of an RNA polymerase along a DNA template. The potential of this technology includes a greater molecular understanding of cellular decision making processes, leading to enhanced opportunities to intervene in disease progression through use of novel treatment modalities. PMID- 15780980 TI - Characterization of Cdk9(55) and differential regulation of two Cdk9 isoforms. AB - Positive transcription elongation factor b (P-TEFb) controls the fraction of initiated RNA polymerase II molecules that make full length transcripts. This important factor is a heterodimer of cyclin-dependent kinase 9 (Cdk9) and one of four cyclin partners, cyclin T1, T2a, T2b or K. There are two isoforms of Cdk9 in mammalian cells, Cdk9(42) and Cdk9(55). Cdk9(55) has a 117 residue amino terminal extension not present in Cdk9(42). An expression vector with a tetracycline responsive promoter driving FLAG-tagged Cdk9(55) and a HeLa 37 Tet-Off cell line were constructed. FLAG-tagged Cdk9(55) was inducibly expressed and was found to be localized to the nucleus by immunofluorescence. Western analysis of murine tissues showed that the relative abundance of the two forms of Cdk9 varied across different tissues with liver having more Cdk9(55) than Cdk9(42). During adaptation of primary rat hepatocytes to culture the ratio of the two forms of Cdk9 changed. Initially, Cdk9(55) was the predominate form, but as the cells began to enter the cell cycle Cdk9(42) became the major form. During this change, expression of Cdk9(42) was induced, while Cdk9(55) remained relatively constant. PMID- 15780981 TI - Mitochondrial genome of the Chinese mitten crab Eriocheir japonica sinenesis (Brachyura: Thoracotremata: Grapsoidea) reveals a novel gene order and two target regions of gene rearrangements. AB - We determined the complete 16,354 bp mitochondrial DNA sequence of the Chinese mitten crab Eriocheir japonica sinesnesis. It consists of 13 protein coding genes (PCGs), 2 rRNAs, and 22 tRNAs, typical of metazoan mitochondrial genomes. With respect to the ancestral crustacean mt gene order, a PCG, the rRNAs, and 12 of the tRNAs appear to be rearranged. This rearrangement is novel in the arthropods and suggests an accelerated rate of mt genome rearrangement in this brachyuran lineage based on the relative rate of gene rearrangement. It is typical in arthropods that all of the rearranged genes or gene blocks take place at both nad3-nad5 and nad5-nad4 gene junctions. Such occurrence additionally revealed two target regions of frequent rearrangement in mitochondrial genomes of decapods, even in that of the non-hexapod arthropods according to our comparative studies among 32 taxa. Additionally, selective constraint on sharing the single introducing location is apparent for most of the rearrangements that occurred at the nad3-nad5 gene junction of these taxa. The gene arrangement features at both gene junctions allow the reconstruction of relationships among the advanced decapods. These features are therefore characteristic molecular markers in phylogenetic inference. The genomic organization differences at both gene junctions provide new evidence of extremely divergence between Heterotremata and Thoracotremata in brachyuran crabs. A duplication/loss (random and nonrandom) recombination model was proposed to account for the generation of the gene order in E. japonica sinesnesis under the guide of intergenic spacers. PMID- 15780982 TI - PCR-based unidirectional deletion method for creation of comprehensive cDNA libraries. AB - A new strategy for the rapid creation of DNA deletion libraries using a simple PCR-based method is presented. Unidirectional deletion fragments are created and may be cloned into any vector system without the constraint of using restriction enzymes. Our strategy combines methodologies from DNA sequencing, PCR, and homologous recombination (either in vivo or in vitro) to allow for the creation of a library containing fragments representing all possible deletions of a given cDNA. Using this strategy we have successfully constructed a deletion library of the cDNA encoding for the lumenal domain of yeast Ire1p, and have shown that resulting fragments range from 100 bp to the full length cDNA (1557 bp). This method is simple, inexpensive, and can easily be adapted for automated high throughput research. PMID- 15780983 TI - Green T(R) cells. AB - Identification of the transcription factor Foxp3 as a "master regulator" of regulatory T (T(R)) cells was a major discovery. A new study by Fontenot et al. (2005), reported in this issue of Immunity, provides novel insights into T(R) cell biology by tracking their behavior in mice expressing a GFP-Foxp3 fusion protein reporter. PMID- 15780984 TI - Do cytotoxic lymphocytes kill via reactive oxygen species? AB - A paper by Martinvalet et al. (2005) in this issue of Immunity examines the mechanisms used by granzyme A to kill target cells after its cytoplasmic injection by cytotoxic lymphocytes. They show that this protease induces mitochondrial damage and generation of reactive oxygen species that are necessary for cell death. PMID- 15780985 TI - Peripheral tolerance of CD8 T lymphocytes. AB - Whereas high-avidity recognition of peptide-MHC complexes by developing T cells in the thymus results in deletion and promotes self-tolerance, such recognition by mature T cells in the periphery results in activation and clonal expansion. This dichotomy represents the basis of a dilemma that has stumped immunologists for many years, how are self-specific T cells tolerized in the periphery? There appear to be two important criteria used to achieve this goal. The first is that in the absence of inflammatory pathogens, tolerance is promoted when T cells recognize antigen presented by quiescent dendritic cells (DCs) expressing low levels of costimulatory molecules. A second critical factor that defines "self" and drives tolerance through deletion, anergy, or suppression is the persistence of antigen. PMID- 15780986 TI - Phagocytosis of apoptotic neutrophils regulates granulopoiesis via IL-23 and IL 17. AB - Homeostatic regulation of neutrophil production is thought to match neutrophil elimination to maintain approximately constant numbers in the blood. Here, we show that IL-17, a cytokine that regulates granulopoiesis through G-CSF, is made by gammadelta T cells and unconventional alphabeta T cells. These neutrophil regulatory T cells (Tn) are expanded in mice that lack leukocyte adhesion molecules, which have neutrophilia and defective neutrophil trafficking. Normal neutrophils migrate to tissues, where they become apoptotic and are phagocytosed by macrophages and dendritic cells. This curbs phagocyte secretion of IL-23, a cytokine controlling IL-17 production by Tn cells. Adoptive transfer of wild type, but not adhesion molecule-deficient, neutrophils into mice deficient in beta2 integrins transiently decreases neutrophilia and reduces levels of serum IL 17. Antibody blockade of the p40 subunit of IL-23 reduces neutrophil numbers in wild-type mice. These findings identify a major homeostatic mechanism for the regulation of neutrophil production in vivo. PMID- 15780987 TI - A human CD34(+) subset resides in lymph nodes and differentiates into CD56bright natural killer cells. AB - In humans, T cells differentiate in thymus and B cells develop in bone marrow (BM), but the natural killer (NK) precursor cell(s) and site(s) of NK development are unclear. The CD56bright NK subset predominates in lymph nodes (LN) and produces abundant cytokines compared to the cytolytic CD56dim NK cell that predominates in blood. Here, we identify a novel CD34dimCD45RA(+) hematopoietic precursor cell (HPC) that is integrin alpha4beta7bright. CD34dimCD45RA(+)beta7bright HPCs constitute <1% of BM CD34(+) HPCs and approximately 6% of blood CD34(+) HPCs, but >95% of LN CD34(+) HPCs. They reside in the parafollicular T cell regions of LN with CD56bright NK cells, and when stimulated by IL-15, IL-2, or activated LN T cells, they become CD56bright NK cells. The data identify a new NK precursor and support a model of human NK development in which BM-derived CD34dimCD45RA(+)beta7bright HPCs reside in LN where endogenous cytokines drive their differentiation to CD56bright NK cells in vivo. PMID- 15780988 TI - Nonredundant roles of Sema4A in the immune system: defective T cell priming and Th1/Th2 regulation in Sema4A-deficient mice. AB - The class IV semaphorin Sema4A provides a costimulatory signal to T cells. To investigate the possible developmental and regulatory roles of Sema4A in vivo, we generated Sema4A-deficient mice. Although Sema4A-deficient mice develop normally, DCs and T cells from knockout mice display poor allostimulatory activities and T helper cell (Th) differentiation, respectively. Interestingly, in addition to its expression on DCs, Sema4A is upregulated on Th1-differentiating cells, and it is necessary for in vitro Th1 differentiation and T-bet expression. Consequently, in vivo antigen-specific T cell priming and antibody responses against T cell dependent antigens are impaired in the mutant mice. Additionally, Sema4A deficient mice exhibit defective Th1 responses. Furthermore, reconstitution studies with antigen-pulsed DCs reveal that DC-derived Sema4A is important for T cell priming, while T cell-derived Sema4A is involved in developing Th1 responses. Collectively, these results indicate a nonredundant role of Sema4A not only in T cell priming, but also in the regulation of Th1/Th2 responses. PMID- 15780989 TI - Dual functions of Runx proteins for reactivating CD8 and silencing CD4 at the commitment process into CD8 thymocytes. AB - To understand how CD8 expression is regulated during the transition process from CD4+8+ (CD4 and CD8 double positive, DP) to CD4-8+ (CD8 single positive, CD8SP) cells in the thymus, the involvement of Runx proteins in the alteration of chromatin configuration was investigated. Using the chromatin immunoprecipitation assay, we first demonstrated that Runx proteins bind to the stage-specific CD8 enhancer, as well as the CD4 silencer, in CD8SP thymocytes. Among Runx family members, Runx3 expression was initiated in DP thymocytes receiving a positive selection signal and increased in concert with differentiation to the CD8SP stage. Furthermore, reactivation of the CD8 gene, as well as CD4 silencing, was suppressed in positively selected thymocytes of Runx dominant-negative transgenic mice. These results suggest that Runx proteins, especially Runx3, are involved in lineage specification of CD8 T cells and provide important information for understanding the mechanism for the mutually exclusive expression of coreceptors in mature thymocytes. PMID- 15780990 TI - Regulatory T cell lineage specification by the forkhead transcription factor foxp3. AB - Regulatory T cell-mediated dominant tolerance has been demonstrated to play an important role in the prevention of autoimmunity. Here, we present data arguing that the forkhead transcription factor Foxp3 acts as the regulatory T cell lineage specification factor and mediator of the genetic mechanism of dominant tolerance. We show that expression of Foxp3 is highly restricted to the subset alphabeta of T cells and, irrespective of CD25 expression, correlates with suppressor activity. Induction of Foxp3 expression in nonregulatory T cells does not occur during pathogen-driven immune responses, and Foxp3 deficiency does not impact the functional responses of nonregulatory T cells. Furthermore, T cell specific ablation of Foxp3 is sufficient to induce the identical early onset lymphoproliferative syndrome observed in Foxp3-deficient mice. Analysis of Foxp3 expression during thymic development suggests that this mechanism is not hard wired but is dependent on TCR/MHC ligand interactions. PMID- 15780991 TI - Rgs1 and Gnai2 regulate the entrance of B lymphocytes into lymph nodes and B cell motility within lymph node follicles. AB - Signaling by G protein-coupled receptors coupled to Galpha(i) assists in triggering lymphocyte movement into and out of lymph nodes. Here, we show that modulating the signaling output from these receptors dramatically alters B cell trafficking. Intravital microscopy of adoptively transferred B cells from wild type and Rgs1-/- mice revealed that Rgs1-/- B cells stick better to lymph node high endothelial venules, home better to lymph nodes, and move more rapidly within lymph node follicles than do wild-type B cells. In contrast, B cells from Gnai2-/- mice enter lymph nodes poorly and move more slowly than do wild-type B cells. The Gnai2-/- mice often lack multiple peripheral lymph nodes, and their B cells respond poorly to chemokines, indicating that Galpha(i1) and Galpha(i3) poorly compensate for the loss of Galpha(i2). These results demonstrate opposing roles for Rgs1 and Gnai2 in B cell trafficking into and within lymph nodes. PMID- 15780992 TI - Granzyme A induces caspase-independent mitochondrial damage, a required first step for apoptosis. AB - Granzyme A (GzmA) triggers cell death with apoptotic features by targeting the endoplasmic reticulum-associated SET complex, which contains the GzmA-activated DNase NM23-H1, its inhibitor SET, and Ape1. The SET complex was postulated to translocate to the nucleus in response to oxidative stress and participate in its repair. Because mitochondrial damage is important in apoptosis, we investigated whether GzmA damages mitochondria. GzmA induces a rapid increase in reactive oxygen species and mitochondrial transmembrane potential loss, but does not cleave bid or cause apoptogenic factor release. The mitochondrial effect is direct, does not require cytosol, and is insensitive to bcl-2 and caspase inhibition. SET complex nuclear translocation, which occurs within minutes of peroxide or GzmA treatment, is dependent on superoxide generation since superoxide scavengers block it. Superoxide scavengers also block apoptosis by CTLs expressing GzmA and/or GzmB. Therefore, mitochondrial damage is an essential first step in killer cell granule-mediated pathways of apoptosis. PMID- 15780993 TI - Primary antitumor immune response mediated by CD4+ T cells. AB - Gene-targeted mice have recently revealed a role for lymphocytes and interferon gamma (IFNgamma) in conferring protection against cancer, but the mechanisms remain unclear. Here, we have characterized a successful primary antitumor immune response initiated by naive CD4+ T cells. Major histocompatibility complex class II (MHC-II)-negative myeloma cells injected subcutaneously into syngeneic mice were surrounded within 3 days by macrophages that captured tumor antigens. Within 6 days, naive myeloma-specific CD4+ T cells became activated in draining lymph nodes and subsequently migrated to the incipient tumor site. Upon recognition of tumor-derived antigenic peptides presented on MHC-II by macrophages, the myeloma specific CD4+ T cells were reactivated and started to secrete cytokines. T cell derived IFNgamma activated macrophages in close proximity to the tumor cells. Tumor cell growth was completely inhibited by such locally activated macrophages. These data indicate a mechanism for immunosurveillance of MHC-II-negative cancer cells by tumor-specific CD4+ T cells through collaboration with macrophages. PMID- 15780994 TI - Defective central tolerance induction in NOD mice: genomics and genetics. AB - The genetic determinism of type-1 diabetes in NOD mice likely involves complementary defects in central T cell tolerance induction and peripheral immunoregulation. To study the contribution of the NOD genetic background to central tolerance, we followed the behavior of BDC2.5 clonotype thymocytes in fetal thymic organ cultures (FTOC). The NOD genetic background encodes a quantitative deficiency in the ability to delete these self-reactive thymocytes and to divert them to the CD8alphaalpha lineage. In genetic analyses, comparing NOD and B6.H2g7 FTOCs, the NOD defect incorporated the influence of several loci (notably ones on chr1 and 3). Microarray analyses assessing FTOCs from the same two strains argued that the NOD abnormality reflects the combined effects of turning down the gene expression program that provokes apoptosis and turning on a new program promoting cell survival. Intersection of the data from the two approaches points to a small set of attractive candidate genes. PMID- 15780995 TI - Interaction and electron transfer between the high molecular weight cytochrome and cytochrome c3 from Desulfovibrio vulgaris Hildenborough: kinetic, microcalorimetric, EPR and electrochemical studies. AB - The complex formation between the tetraheme cytochrome c3 and hexadecaheme high molecular weight cytochrome c (Hmc), the structure of which has recently been resolved, has been characterized by cross-linking experiments, EPR, electrochemistry and kinetic analysis, and some key parameters of the interaction were determined. The analysis of electron transfer between [Fe] hydrogenase, cytochrome c3 and Hmc demonstrates a redox-shuttling role of cytochrome c3 in the pathway from hydrogenase to Hmc, and shows an effect of redox state on the interaction between the two cytochromes. The role of polyheme cytochromes in electron transfer from periplasmic hydrogenase to membrane redox proteins is assessed. A model with cytochrome c3 as an intermediate between hydrogenase and various polyheme cytochromes is proposed and its physiological consequences are discussed. PMID- 15780996 TI - Mechanism of horseradish peroxidase-catalyzed heme oxidation and polymerization (beta-hematin formation). AB - Horseradish peroxidase (HRP) catalyzes the polymerization of free heme (beta hematin formation) through its oxidation. Heme when added to HRP compound II (FeIV=O) causes spectral shift from 417 nm (Compound II) to 402 nm (native, FeIII) indicating that heme may be oxidized via one-electron transfer. Direct evidence for one-electron oxidation of heme by HRP intermediates is provided by the appearance of an E.s.r signal of a 5,5-dimethyl-1-pyrroline N-oxide (spin trap)-heme radical adduct (a1H=14.75 G, a2H=4.0 G) in E.s.r studies. Heme polymerization by HRP is inhibited by spin trap indicating that one-electron oxidation product of heme ultimately leads to the formation of heme-polymer. HRP, when incubated with diethyl pyrocarbonate (DEPC), a histidine specific reagent, shows concentration dependent loss of heme-polymerization indicating the role of histidine residues in the process. We suggest that HRP catalyzes the formation of heme-polymer through one-electron oxidation of free heme. PMID- 15780997 TI - Productive interaction of chaperones with substrate protein domains allows correct folding of the downstream GFP domain. AB - Green fluorescent protein (GFP) has been used to report protein folding by correlating solubility with fluorescence. In a GFP fusion protein, an upstream aggregation-prone domain can disrupt de novo folding of the GFP domain in Escherichia coli, resulting in a loss of fluorescence. Previously, we showed that prevention of misfolding of the upstream aggregation-prone domain by a coupled folding and binding interaction during protein synthesis restored both GFP fluorescence and solubility. Since molecular chaperones often fold nascent polypeptides through a bind-and-release interaction, the question remains whether the chaperone interaction with the upstream aggregation-prone domain enhances GFP fluorescence. Here, we demonstrate that a significant increase in GFP fluorescence occurred only when appropriate chaperones that recognized the aggregation-prone protein and helped its folding were co-expressed. A possible correlation between GFP fluorescence and the productive folding by chaperones is proposed. This study may provide a general strategy for identifying chaperones specific for difficult-to-fold proteins. PMID- 15780999 TI - The discovery of four distinct glutamate dehydrogenase genes in a strain of Halobacterium salinarum. AB - In earlier work, two glutamate dehydrogenase (GDH) proteins were purified from a strain of the halophilic archaeon Halobacterium salinarum (NRC-36014). One of these, an NAD+-specific enzyme, was matched to a cloned gene from H. salinarum (GenBank accession number: X63837 S75579) by sequencing peptide fragments. Analysis of enzymatic digests of the NADP+-GDH and database searching have now established that a gene encoding this protein exists in the full genomic sequence of Halobacterium sp. NRC-1 as gdhA1, together with two other distinct gdh genes, gdhA2 and gdhB. From N-terminal sequence, it is clear that the genomic listing incorrectly assigns the start codon for gdhA1 and the corresponding protein is 43 amino acids longer than previously indicated. The three genes could be amplified by PCR either from NRC-1, as expected, or from NRC-36014 (GenBank accession numbers: YA840085-AY840087). A gene encoding the previously purified NAD+-GDH, is absent from the NRC-1 genome but can be successfully amplified from genomic DNA of NRC-36014 (GenBank accession number: AY840088). This establishes that NRC 36014 contains four gdh genes. PMID- 15780998 TI - Gene characterization and predicted protein structure of the mitochondrial chaperonin HSP10 of Trypanosoma cruzi. AB - Heat shock protein (HSP) 10 is a member of the highly conserved group of molecular chaperons, which are necessary for efficient folding of many proteins in normal and stress conditions and have been implicated in several human diseases. We have characterized the HSP10 genes of Trypanosoma cruzi, the causative agent of Chagas' disease. After sequence analysis of clones obtained from the T. cruzi Genome Initiative, we show that the T. cruzi HSP10 coding region is 300 bp long, encoding a polypeptide of 100 amino acids with highest sequence identity (83%) to HSP10 of Trypanosoma brucei and lowest (28%) to HSP10 of Escherichia coli. The T. cruzi HSP10 genes are arranged in 3 tandemly repeated copies, which give rise to a major mRNA of 1.0 kb that remains unaltered during heat shock; a smaller mRNA species is induced at 37 degrees C by alternate polyadenylation. Finally, the presence of a conserved 5-amino acid residue deletion in trypanosomatid HSP10s led us to generate a molecular model of the T. cruzi HSP10 structure. The oligomeric assembly of this model shows some peculiar characteristics that may have functional significance. PMID- 15781000 TI - Molecular cloning and functional characterization of two apple S adenosylmethionine decarboxylase genes and their different expression in fruit development, cell growth and stress responses. AB - Two full-length S-adenosylmethionine decarboxylase (SAMDC) cDNAs, MdSAMDC1 and MdSAMDC2, were isolated from apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. Both cDNAs encoded tiny and small ORFs in addition to the SAMDC ORFs, and genomic sequences of MdSAMDC1 and MdSAMDC2 contained two or three introns in the 5' upstream regions, respectively. Yeast complementation experiment indicated that two MdSAMDCs encoded functional proteins, and that the tiny and small ORFs possibly repressed their translation efficiency. RNA gel blot analysis showed that MdSAMDC1 were differentially regulated in fruits depending on the developmental stage and in cell suspension during the culture period, but MdSAMDC2 did not. In contrast, MdSAMDC2 was positively induced by cold and salt stresses, but MdSAMDC1 was not. These results suggest that MdSAMDC1 is mainly involved in fruit development and cell growth while MdSAMDC2 in stress responses, compared with their respective counterpart. PMID- 15781001 TI - Circulating amounts of osteoprotegerin and RANK ligand: genetic influence and relationship with BMD assessed in female twins. AB - Osteoprotegerin (OPG) is a circulating receptor that inhibits osteoclastogenesis by binding to RANK ligand (RANKL). OPG knock-out animals develop severe osteoporosis. Treatment with OPG lowers bone resorption and increases BMD. OPG production is influenced by a wide range of hormones and cytokines. The influence of genetic factors on circulating amounts of OPG and RANKL is not known. BMD has been demonstrated to have a high heritability and there is evidence also that bone turnover and bone loss rates are controlled at least in part by genetic factors. OBJECTIVE: Assessing the genetic impact on serum OPG and RANKL in women and estimation of the relative contribution of this inheritance to the total heritability of BMD. METHODS: 188 female twins (52 DZ and 42 MZ pairs) from the Danish Twin Registry were included in the study. Mean age was 35 years (range 19 64 years), average spine BMD was 1.04 +/- 0.11 g/cm2. Serum levels of OPG and RANKL were measured by ELISA (Biomedica, Vienna, Austria). This register covers twins born in Denmark since 1870. Heritability and environmental influence was assessed using a maximum-likelihood model for genetic pleiotropy. RESULTS: RANKL levels showed a negative correlation with age and lower values in smokers. OPG levels were higher in postmenopausal women. Heritability (h(2)) was 85% for spine BMD and 52% for serum RANKL after adjustment for age, smoking and BMI. By contrast, there was no significant genetic influence on OPG levels (h(2) = 0, 95% CI: 0 to 0.31). Serum OPG was determined almost exclusively by individual environment (e(2) = 0.79), with a small, non-significant contribution from shared environment (c(2) = 0.21). Restricting analyses to the 158 premenopausal twins did not alter the findings. CONCLUSIONS: Serum OPG and RANKL levels have only a weak relation to BMD in healthy women. Phenotype correlations indicate that the genes that contribute to twin similarity for BMD are not genes regulating serum levels of RANKL or OPG. The weak correlation with BMD appears to consist in shared environmental factors. PMID- 15781002 TI - Modulation of CYP27B1 and CYP24 mRNA expression in bone is independent of circulating 1,25(OH)2D3 levels. AB - Circulating levels of 1,25-dihydroxyvitamin D (1,25D) are determined by bioactivation catalyzed by the renal 25-hydroxyvitamin D 1alpha-hydroxylase (CYP27B1) and degradation through the action of the renal 25-hydroxyvitamin D 24 hydroxylase (CYP24). CYP27B1 and CYP24 are also present in bone cells, but little is known of their physiological role. The purpose of this study was to determine the changes that occur with aging on the expression of CYP27B1 and CYP24 mRNA in whole kidney and femora of female Sprague-Dawley rats. Real-time RT-PCR was used to measure CYP27B1, CYP24 and vitamin D receptor (VDR) mRNA levels in the kidneys and bones of animals aged between 3 weeks and 2 years. Circulating 1,25D levels decreased exponentially with age which was correlated with both reduced kidney CYP27B1 mRNA (R(2) = 0.72) and increased CYP24 mRNA levels (R(2) = 0.71). In the bone, CYP27B1 mRNA levels were maintained at their highest level throughout the ages of 3 to 15 weeks before decreasing in adult animals (P < 0.05). Bone CYP24 mRNA levels were positively correlated with bone CYP27B1 mRNA and not circulating 1,25D levels (R(2) = 0.74). Levels of bone CYP27B1 mRNA were positively correlated with distal femoral epiphyseal trabecular number (Tb.N) (R(2) = 0.74) and negatively with the trabecular thickness (Tb.Th) (R(2) = 0.56) in animals aged between 12 weeks and 2 years. These findings indicate that the regulation of CYP27B1 and CYP24 mRNA expression in the bone is unique from that in the kidney. The synthesis of 1,25D in bone tissue regulates bone CYP24 expression and is associated with bone mineralization suggesting that vitamin D metabolism has an autocrine or paracrine function. PMID- 15781003 TI - In vitro differentiation profile of osteoblasts derived from patients with Saethre-Chotzen syndrome. AB - Seathre-Chotzen syndrome (SCS) is an autosomal dominant craniosynostosis syndrome, associated with loss-of-function mutations in the basic helix-loop helix transcription factor, TWIST1. The biologic activity of TWIST1 has been implicated in the inhibition of differentiation of multiple cell lineages. Therefore, premature fusion of cranial sutures (craniosynostosis) in SCS may be mediated by altered differentiation of calvarial osteoblasts. In this study, we evaluated osteoblasts derived from calvarial bone of three patients with SCS and three unaffected individuals as controls to investigate the principle stages of osteoblast differentiation: (1) proliferation, (2) matrix maturation, and (3) mineralization. Using a BrdU-Hoechst flow cytometry assay, we found that the percent of proliferating cells was significantly reduced in cells derived from patients with SCS compared with those derived from controls (P < or = 0.05). In the matrix maturation stage, alkaline phosphatase (ALP) enzyme activity and the expression of extracellular matrix genes, collagen I alpha 2 (COL1A2), osteopontin (OPN), osteocalcin (OC), and the runt-related transcription factor RUNX2 were examined by enzymatic assay and real-time quantitative RT-PCR, respectively. We identified no significant differences in the expression of matrix related transcripts. However, we found significant reductions in ALP activity on days 3 and 7 and in RUNX2 expression on days 14 and 21 (P < or = 0.05). Quantitative alizarin red S mineralization assays showed a trend toward increased mineralization in osteoblasts derived from patients with SCS at days 21 and 28, although not statistically significant. Our results demonstrated that loss-of-function mutations of TWIST1 led to reduced proliferation regardless of the functional domain affected. We did not find any conclusive differences in matrix maturation or mineralization in these primary osteoblasts. It is plausible that mutations in different functional domains of TWIST1 have divergent effects on these later stages of differentiation. PMID- 15781004 TI - Identification of differentially expressed genes between osteoarthritic and normal trabecular bone from the intertrochanteric region of the proximal femur using cDNA microarray analysis. AB - Osteoarthritis (OA) is a common age-related joint disease resulting in progressive degenerative damage to articular cartilage. The etiology of primary OA has not yet been determined. However, there is evidence supporting the hypothesis that primary OA is a disease affecting bone remodeling in addition to articular cartilage. In this study, we have used cDNA microarray analysis to compare gene expression in bone between normal (CTL) and OA individuals. Trabecular bone was sampled from the intertrochanteric region of the proximal femur, a site distal to the diseased hip joint. Total RNA was extracted from three pairs of age- and sex-matched CTL and OA bone samples, reverse-transcribed and radioactively labeled to generate cDNA probes, before hybridization with the Research Genetics GF211 human gene microarray filter. The CTL and OA samples were found to have similar levels of gene expression for more than 4000 known human genes. However, forty-one genes were identified that were differentially expressed, twofold or more, between all three CTL-OA sample pairs. Using semi quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis, three genes, fms-like tyrosine kinase 1 (FLT1), plexin B1 (PLXNB1), and small inducible cytokine A2 (SCYA2), were confirmed to be consistently expressed at lower levels in OA, in a majority of twenty age- and sex-matched CTL-OA bone sample pairs tested. FLT1, PLXNB1, and SCYA2 have known or potential roles in angiogenesis and bone remodeling. Down-regulation of these genes is consistent with a role for bone in the pathogenesis of OA. PMID- 15781005 TI - Genetic and environmental determinants of bone mineral density in Chinese women. AB - BMD is a complex trait determined by genetic and lifestyle factors. To assess the genetic and environmental determinants of BMD in southern Chinese women, we studied a community-based cohort of 531 pre- and postmenopausal southern Chinese women and assessed the influence of 12 candidate gene loci and lifestyle risk factors on spine and hip BMD. The candidate genes studied include estrogen receptor alpha (ESR1) and beta (ESR2), calcium sensing receptor (CASR), vitamin D receptor (VDR), collagen type Ialpha1 (COLIA1), and LDL receptor-related protein 5 (LRP5). Social, medical, reproductive history, dietary habits and lifestyle factors were determined using a structured questionnaire. Single nucleotide polymorphisms (SNPs) of the COLIA1 and LRP5 gene in Chinese were determined by direct sequencing. Nucleotide (nt) -1363C/G and -1997 G/T of COLIA1, nt 266A/G, 2220C/T and 3989C/T of LRP5 gene were analyzed. Using stepwise multiple linear regression analyses, body weight was the strongest predictor for BMD in premenopausal women (n = 262), which accounted for 15.9% of the variance at the spine, 20% at femoral neck, 17.1% at trochanter, 24.3% at total hip and 10.9% at the Ward's triangle. Other significant predictors were ESR1 Ivs1-397T/C genotype (2.2% at the spine); LRP5 2220C/T genotype (1.3% at the spine, 1.6% at the trochanter); LRP5 266A/G genotype (1.1% at Ward's triangle); age at menarche (1.3% at trochanter) and age (2.0% at Ward's triangle). As for postmenopausal women (n = 269), body weight ( approximately 25% at various sites) and age (approximately 16% at femoral neck, trochanter, total hip and Ward's triangle sites) were the strongest predictors of BMD. Other significant predictors were age at menarche (4.4% at spine, 0.7% at femoral neck, 1.4% at trochanter, and 1.4% at Ward's triangle); weight bearing physical activity (2.1% at trochanter and 1% at total hip); calcium intake (1.1% at femoral neck, 0.9% at trochanter, and 1.7% at total hip) ; height (0.7% at trochanter); and ESR2 1082A/G genotype (0.8% at trochanter). We conclude that BMD at various sites and at different time span of a woman is modified by different genetic and lifestyle factors, suggesting that BMD is highly dependent on gene-environmental interactions. PMID- 15781006 TI - Human parathyroid hormone (1-34) accelerates the fracture healing process of woven to lamellar bone replacement and new cortical shell formation in rat femora. AB - This study aimed to test whether intermittent treatment of human parathyroid hormone [hPTH(1-34)] disturbs or accelerates the fracture healing process using rat surgical osteotomy model. One hundred five, 5-week-old SD rats were allocated to vehicle control (CNT) and four PTH groups; 10 and 30 microg/kg of hPTH(1-34) treatment before surgery (P10, P30), and treatment before and after surgery (C10, C30). All animals were given subcutaneous injections three times a week for 3 weeks. Then, fractures were produced by transversely cutting the midshaft of bilateral femora and fixing with intramedullary wire. Human PTH(1-34) treatment was continued in C10 and C30 groups until sacrifice at 3, 6, and 12 weeks after surgery. The femora were assessed by peripheral quantitative computed tomography, three-point bending mechanical test, and histomorphometry. Total cross-sectional area was not significantly different among all groups at any time point. At 3 weeks after surgery, the lamellar bone/callus area was significantly increased in C10 and C30 groups compared to the other groups. At 6 weeks, remodeling of woven bone to lamellar bone in the callus was almost complete in all groups. At 12 weeks, percent new cortical shell area was significantly higher in C10 and C30 groups compared to the other groups, and the ultimate load in mechanical testing was significantly higher in C30 group than in CNT, P10, and P30 groups. Intermittent PTH treatment at 30 microg/kg before and after osteotomy accelerated the healing process as evidenced by earlier replacement of woven bone to lamellar bone, increased new cortical shell formation, and increased the ultimate load up to 12 weeks after osteotomy. PMID- 15781007 TI - Homocysteine as a predictive factor for hip fracture in stroke patients. AB - Risk of hip fractures in stroke patients is higher than that in a reference population. Hyperhomocysteinemia is regarded as a risk factor for ischemic stroke. The high prevalence of osteoporosis among patients with homocystinuria suggests that hyperhomocysteine may also increase the risk of fractures. To determine the association between homocysteine concentration and the risk of hip fractures, we studied a cohort of stroke patients with hemiplegia. Age-adjusted incidence rates of a hip fracture were calculated for quartiles of homocysteine concentrations. Cox proportional-hazard regression was used to calculate hazard ratios for quartiles of homocysteine levels. The initial enrolment of 433 hemiplegic patients with ischemic stroke, older than 65 years old, were followed for up to 10 years. The mean plasma homocysteine concentration at the enrolment was 14.1 +/- 5.2 micromol/L. There were 33 hip fractures among men and 46 among women during the mean follow-up period of 9.0 years. The age-adjusted incidence rates per 1000 person-years for hip fractures increased almost linearly from 2.89 in the lowest to 27.87 in the highest quartiles of homocysteine levels. We conclude that hyperhomocysteinemia is one of the risk factors for hip fractures in stroke patients. PMID- 15781008 TI - Usefulness of ventricular repolarization dynamicity in predicting arrhythmic deaths in patients with ischemic cardiomyopathy (from the European Myocardial Infarct Amiodarone Trial). AB - The European Myocardial Infarct Amiodarone Trial (EMIAT) investigated the effects of amiodarone versus placebo in patients after myocardial infarction who had left ventricular ejection fraction /=1 stenosis of >/=70%) without AMI at hospital presentation was evaluated. Baseline demographics, cardiac risk factors, clinical presentation, therapeutic procedures, and discharge medications were recorded. Patients were followed for a mean of 3.0 +/- 1.9 years (range 1 month to 6.9 years) for outcomes of all-cause death or AMI. Patients' average age was 65 +/- 11 years and 77% were men. Overall, 10% died and 5% had a nonfatal AMI. Discharge beta-blocker prescription was associated with an increased event-free AMI survival rate for all-cause death (no beta blocker 88.3%, beta blocker 94.5%, p <0.001) and death/AMI (no beta blocker 83.4%, beta blocker 89.2%, p <0.001) but not non-fatal AMI (no beta blocker 93.6%, beta blocker 94.1%, p = 0.60). After adjustment for 16 covariates, including statin prescription, angiotensin-converting enzyme inhibitor prescription, and type of baseline therapy, the effect of beta blockers on the combination end point of death/AMI was eliminated. However, the effect of beta blockers on death remained (hazard ratio 0.66, 95% confidence interval 0.47 to 0.93, p = 0.02). Thus, beta blockers are clearly indicated for most patients who have HF or AMI, and our results suggest that patients who have coronary artery disease without these conditions have approximately the same protective benefit against death. No effect was observed on longitudinal incidence of AMI or the combination of death/nonfatal MI. PMID- 15781010 TI - Doppler-derived left ventricular filling pressures and the regulation of heart rate recovery after exercise in patients with suspected coronary artery disease. AB - Slowed heart rate (HR) recovery after exercise is strongly predictive of increased long-term mortality. The factors responsible for impaired HR regulation are not fully understood. We performed echocardiography with tissue Doppler imaging in 121 patients before maximal exercise testing. HR recovery was measured 1 minute after the end of exercise in the supine position. The best echocardiographic correlate of HR recovery was the ratio of early mitral flow velocity (E) to early diastolic mitral annular velocity (Ea; r = -0.781, p <0.001). This correlation was not affected by the use of negative chronotropic agents. Patients whose E/Ea was <10 had a faster 1-minute HR recovery and a greater chronotropic response during exercise than did those whose E/Ea was >/=10. Receiver-operator characteristic analysis showed that an E/Ea >/=10.3 predicted 1-minute HR recovery of /=2 of the following: men >/=45 years, women >/=55 years, hypertension, high density lipoprotein cholesterol <40 mg/dl, total cholesterol >/=200 mg/dl, or obesity. Lipid treatment rates were estimated among those needing therapy (defined as low-density lipoprotein cholesterol >/=130 mg/dl or currently receiving lipid-lowering therapy). The overall lipid-lowering therapy treatment rate was 38% and the estimated lipid treatment gap (percent needing treatment who were not receiving it) was 62%. PMID- 15781018 TI - Relation of serum uric acid with metabolic risk factors in asymptomatic middle aged Brazilian men. AB - This study in 352 asymptomatic middle-aged Brazilian men demonstrated that serum uric acid increases linearly with an increasing number (0 to >/=3) of metabolic risk factors (5.78 +/- 1.1, 6.14 +/- 1.0, 6.27 +/- 1.1, and 6.79 +/- 1.3, p <0.001). In patients who had >/=3 metabolic risk factors, there was a higher prevalence of serum uric acid in the highest quartile (7.2 to 10.3 mg/dl) than in the lowest quartile (2.6 to 5.4 mg/dl, 35% vs 12%, p <0.001). Mean serum levels of uric acid were higher in those who had an abnormal ratio of >/=3 for triglyceride to high-density lipoprotein (suggesting insulin resistance) than in those who had a normal ratio (6.6 +/- 1.2 vs 5.87 +/- 1 mg/dl, p <0.001). PMID- 15781019 TI - Effect of docosahexaenoic acid on lipoprotein subclasses in hyperlipidemic children (the EARLY study). AB - To test the hypothesis that a dietary omega-3 fatty acid, docosahexaenoic acid, improves the lipoprotein subclass profile of children who have hyperlipidemia, we conducted a randomized, double-blind, placebo-controlled study. Children who had hyperlipidemia (n = 20) were stabilized on a low-fat diet for 6 weeks and then randomized to receive 1.2 g/day of docosahexaenoic acid for 6 weeks or placebo. Supplementation with docosahexaenoic acid significantly increased low-density lipoprotein subclass 1 and high-density lipoprotein subclass 2 (large and buoyant; less atherogenic particles) by 91% and 14%, respectively, compared with the placebo phase. Low-density lipoprotein subclass 3 (small and dense; more atherogenic particles) decreased by 48%. PMID- 15781020 TI - Anatomic substrate, procedural results, and clinical outcome of ultrasound-guided left atrial-pulmonary vein disconnection for treatment of atrial fibrillation. AB - This report summarizes the efficacy, safety, and feasibility of intracardiac ultrasound (ICUS) and local electrographic-guided pulmonary vein (PV)-left atrial disconnection, including the isolation of common PV trunks accomplished in 170 consecutive patients with atrial fibrillation (AF). A left common PV was found in 43% of patients with AF. During a follow-up of 549 +/- 330 days after ablation, the AF-free survival rate was 80% and comparable in paroxysmal and persistent AF. PV stenosis was detected in only 1 asymptomatic patient, who required no intervention. PMID- 15781021 TI - Differentiating atrioventricular nodal reentrant tachycardia from tachycardia via concealed accessory pathway. AB - Studies analyzing the diagnostic value of 12-lead electrocardiographic criteria differentiating slow-fast atrioventricular nodal reentrant tachycardia (AVNRT) from atrioventricular reentrant tachycardia (AVRT) due to concealed accessory pathway have shown inconsistent results. In 97 patients (50 with AVNRT, 47 with AVRT) 12-lead electrocardiograms (ECGs) were recorded during sinus rhythm and tachycardia (QRS <120 ms). The ECGs were blinded for diagnosis and patient and analyzed independently by 2 electrophysiologists. The studied criteria differentiating AVNRT from AVRT included pseudo-r'/S, the presence of a retrograde P wave, RP interval, ST-segment depression >/=2 mm with the number and location of the affected leads, QRS amplitude, and cycle length alternans. PMID- 15781022 TI - First experience with a Mobile Cardiac Outpatient Telemetry (MCOT) system for the diagnosis and management of cardiac arrhythmia. AB - Recently, a mobile cardiac outpatient telemetry (MCOT) system has become available that monitors the electrocardiogram continuously, recognizes arrhythmias automatically, and transmits abnormal rhythms instantaneously. MCOT does not require activation by the patient. We report data from the first 100 consecutive patients monitored by this new technology. PMID- 15781024 TI - Value of B-type natriuretic peptide for identifying significantly elevated pulmonary artery wedge pressure in patients treated for established chronic heart failure secondary to ischemic or idiopathic dilated cardiomyopathy. AB - It has been suggested that plasma B-type natriuretic peptide (BNP) level may be used as a noninvasive biomarker of the adequacy of long-term heart failure therapy. The effect of contemporary therapy on the relation between BNP measured using modern commercially available assays and cardiac filling pressures has not been studied in detail, because most of the original studies predate these developments. The investigators sought to assess the diagnostic accuracy of BNP to identify significantly elevated pulmonary capillary wedge pressure (PCWP) in a group of patients with severe chronic heart failure on treatment. BNP correlated well with PCWP (r = 0.50, p <0.001), but the diagnostic accuracy of the test to identify patients with PCWP >15 mm Hg was only 74%, largely because of poor sensitivity or a large number of false-negative test results. Maximizing medical therapy irrespective of plasma BNP results remains the best approach to managing chronic heart failure. PMID- 15781023 TI - Relation of interleukin-6 levels and prothrombin fragment 1+2 to a point-based score for stroke risk in atrial fibrillation. AB - Interleukin-6 levels, but not prothrombin fragment 1 + 2, correlates with a point based score for stroke risk in atrial fibrillation, even after oral anticogulation. PMID- 15781025 TI - Effect of noradrenaline and isoproterenol on lipopolysaccharide-induced tumor necrosis factor-alpha production in whole blood from patients with chronic heart failure and the role of beta-adrenergic receptors. AB - Increased levels of tumor necrosis factor-alpha (TNF-alpha) correlate with poor prognoses in chronic heart failure (CHF). This study demonstrated that noradrenaline and isoproterenol inhibit TNF-alpha production in patients with CHF in ex vivo whole blood in a dose-dependent fashion. The beta-blocker bisoprolol abolishes this effect. PMID- 15781026 TI - Use of cardiac resynchronization therapy to optimize beta-blocker therapy in patients with heart failure and prolonged QRS duration. AB - A retrospective analysis was performed on 52 patients with heart failure to determine the change in beta-blocker therapy after cardiac resynchronization therapy (CRT). After 6 months of CRT, the number of patients receiving beta blocker therapy increased from 36 to 44, with improved clinical outcomes and larger beta-blocker doses, indicating that these 2 therapies may work together to improve outcomes by allowing the use of larger doses of beta blockers while correcting ventricular dyssynchrony. PMID- 15781027 TI - Usefulness of tissue Doppler imaging for estimation of left ventricular filling pressures in patients with systolic and diastolic heart failure. AB - The diagnostic usefulness of the mitral E/E' ratio (derived from tissue Doppler imaging) as an estimate of left ventricular filling pressures was studied in 28 patients with diastolic heart failure (defined by heart failure signs and symptoms but with preserved ejection fraction) and in 46 patients with systolic heart failure (heart failure signs and symptoms and reduced ejection fraction). E/E' was reflective of filling pressures in subjects with diastolic and systolic heart failure and may be of special use in ruling out elevated filling pressures in subjects with suspected diastolic heart failure. PMID- 15781028 TI - Do elderly systolic heart failure patients benefit from beta blockers to the same extent as the non-elderly? Meta-analysis of >12,000 patients in large-scale clinical trials. AB - A meta-analysis of all-cause mortality data involving elderly and non-elderly chronic heart failure patients from 5 completed beta-blocker trials revealed that elderly and non-elderly chronic heart failure patients derived considerable prognostic benefit from beta-blocker therapy without a statistically significant difference in mortality reduction between the 2 groups. PMID- 15781029 TI - Usefulness of serial measurement of N-terminal pro-brain natriuretic peptide plasma levels in asymptomatic patients with aortic stenosis to predict symptomatic deterioration. AB - In 29 initially asymptomatic patients with aortic stenosis followed for an average of 18 months, patients with a N-terminal pro-brain natriuretic peptide (NT-pro-BNP) level above the normal range at baseline were more likely to develop symptoms during follow-up compared with patients with NT-pro-BNP within normal limits. The average increase in NT-pro-BNP per year was greater for patients who developed symptoms compared with patients who remained asymptomatic. Aortic valve area, peak aortic velocity, and the ejection fraction were less reliable predictors of symptom onset. Measurement of NT-pro-BNP in addition to clinical assessment and echocardiography may allow more reliable follow-up and timing of valve replacement for aortic stenosis. PMID- 15781030 TI - Usefulness of a new sound spectral averaging technique to distinguish an innocent systolic murmur from that of aortic stenosis. AB - We present a new method to record and display heart sounds that uses a hand-held computer and stethoscopic recording device. It allows for rapid spectral and waveform displays of murmurs and provides a means for signal averaging of spectral frequency content. Compared with aortic stenosis, innocent murmurs primarily contain frequencies of <300 Hz and persist for a shorter duration at the upper-frequency levels. This method provides for rapid characterization of innocent murmurs, a means for comparison with other murmurs, and transmission of acoustic data to distant sites. PMID- 15781031 TI - Comparison of brain natriuretic peptide and left ventricular diastolic function determined by tissue Doppler in patients with diabetes mellitus, patients with hypertension without diabetes, and in healthy subjects. AB - In this study, it was found that increased plasma B-type natriuretic peptide (BNP) levels in patients with diabetes may be related to left ventricular (LV) diastolic relaxation, independent of LV mass and atherosclerosis (using common carotid intima-media thickness as a surrogate index). A "package of care" of glycemic control and cardiovascular risk management was not associated with reduction in BNP levels. PMID- 15781032 TI - Value of plasma B-type natriuretic peptide as a marker for rejection in pediatric heart transplant recipients. AB - We analyzed 211 consecutive plasma B-type natriuretic peptide (BNP) measurements in 59 pediatric heart transplant patients along with echocardiographic and right ventricular endomyocardial biopsy samples. Patients with a biopsy specimen negative for rejection had significantly lower BNP levels than those patients with a biopsy positive (p /=Pam(2)CAG-Mal-Th approximately Pam(2)CAG-Mal>Pam(3)CAG-Mal-Th (where Th is a HS-peptide) and no appreciable activity was detected for Pam(3)CAG-Mal, Ol(3)CAG-Mal and Ol(3)CAG-Mal-Th. Our findings indicate that subtle modifications in the peptide moiety of lipopeptides have a great impact on the immunomodulatory properties of these molecules. For the engineering of liposome/lipopeptide-based vaccines, the maleimide derivative of Pam(2)CAG appears to be the best candidate. PMID- 15781117 TI - Activation of the lectin pathway in murine lupus nephritis. AB - In systemic lupus erythematosus (SLE), hypocomplementaemia and complement deposition have been described both in man and in experimental models. A major involvement of the classical pathway of complement activation has been demonstrated in this disease, however relatively little is known about the involvement of the lectin pathway. Therefore in the present study we have analyzed the activity of all three pathways of complement activation in murine models of SLE. In the mouse, MBL is expressed in two forms, namely MBL-A and MBL C. In the present study young and old MRL-lpr and control MRL+/+ mice were compared for the levels of complement activity with specific attention for the lectin pathway. It was found that upon aging of both MRL-lpr and MRL+/+ mice, a marked decrease in the activity of the classical pathway (CP) occurs. Levels of alternative pathway (AP) and lectin pathway (LP) activity remain unchanged. Key molecules of these pathways, C1q, C3, MBL-A and MBL-C were analyzed and were all found to be decreased in aged mice of both strains. The levels of MBL-A and MBL-C showed a high degree of correlation and decreased equally. In aged MRL-lpr mice in which autoimmunity is most pronounced, we observed high autoantibody titers and strong deposition of glomerular immune complexes in association with deposition of C1q, C3, MBL-A and MBL-C. In conclusion, these data suggest that in addition to the classical pathway and the alternative pathway also the lectin pathway of complement activation is involved in murine lupus nephritis. PMID- 15781118 TI - A novel molecular mechanism to account for the hygiene hypothesis. PMID- 15781119 TI - Towards a mechanism-based analysis of pharmacodynamic drug-drug interactions in vivo. AB - The combination of drugs is a common practice for enhancing the efficiency of drug treatment, but selection of the optimal combination and the optimal doses remains a matter of trial and error. Prediction of synergistic, additive and antagonistic responses to drug combinations in vivo is therefore of considerable interest. The present review discusses the application of mathematical and statistical models to assess combined drug action by response surface modelling. The most commonly applied models are designed to distinguish between synergistic and additive responses on the basis of a single parameter to indicate whether a drug combination acts synergistic or not. It is, however, recognized that these relatively simple models often do not adequately describe complex drug interactions. This has led to the application of increasingly complex models with multiple drug interaction parameters that can describe a wide range of synergistic and antagonistic responses in a single-response surface. The capability to describe response surfaces with high resolution offers the opportunity to develop an understanding of the mechanisms that underlie the observed combined drug response. Operational models for drug interaction constitute a highly versatile framework for mechanism-based modelling by taking the signal transduction properties of the drug combination into account. On this basis, it is predicted that the occurrence of synergism is favoured by convergence of drug signals late in the signal transduction pathway as opposed to proximal convergence. Furthermore, a high efficiency of signal transduction poses in general a barrier to the occurrence of synergism. The in vivo application of operational models with advanced response surface modelling techniques will facilitate the rational development of synergistic drug combinations. PMID- 15781120 TI - Polymorphisms affecting gene regulation and mRNA processing: broad implications for pharmacogenetics. AB - Functional polymorphisms that alter gene expression and mRNA processing appear to play a critical role in shaping human phenotypic variability. Intensive studies on polymorphisms affecting drug response have revealed multiple modes of altered gene function, frequently involving cis-acting regulatory sequence variants. Experimental and in silico methods have advanced the search for such polymorphisms, but considerable challenges remain. Here, a survey of polymorphisms in drug-related genes indicates that: (a) a substantial proportion of genetic variability still remains unaccounted for; (b) a majority of these genes harbors known regulatory polymorphisms; and (c) a portion of polymorphisms affect splicing and mRNA turnover. Pharmacogenetic optimiziation of individual drug therapy may require a complete understanding of all functional sequence variants in key genes. This review surveys known noncoding polymorphisms in genes encoding cytochrome P450s and other drug-metabolizing enzymes, drug transporters, and drug targets and receptors. Current methods and challenges associated with the identification and characterization of functional polymorphisms are also discussed. PMID- 15781121 TI - Calmodulin kinase signaling in heart: an intriguing candidate target for therapy of myocardial dysfunction and arrhythmias. AB - The multifunctional Ca(2+)/calmodulin (CaM)-dependent protein kinase II (CaMKII) has emerged as a proarrhythmic and procardiomyopathic signal in a wide range of structural heart diseases. This review discusses CaMKII structure and function and recent evidence implicating CaMKII inhibition as a potential strategy for treating myocardial dysfunction and arrhythmias in the setting of structural heart disease. PMID- 15781122 TI - Cardiac nitric oxide: emerging role for nNOS in regulating physiological function. AB - Emerging evidence shows that neuronal nitric oxide synthase (nNOS) plays several diverging roles in modulating cardiac function. This review examines the nitric oxide (NO) pathway and the regulatory mechanisms to which nNOS signalling is sensitive. These mechanisms are diverse and include regulation of gene expression, posttranscriptional regulation, protein trafficking, allosteric modulation of nNOS and redox modification to alter NO bioavailability once synthesised. Functionally, alteration in nNOS-NO signalling in the heart may correlate with different cardiac regulatory states. The idea of this being associated with exercise-trained states and myocardial disease is discussed. PMID- 15781123 TI - Molecular regulation of Th2 immunity by dendritic cells. AB - The interactions between dendritic cells (DCs) and T cells determine the fate of an immune response to pathogenic microbes and to harmless allergens alike. The interactions between DCs and T cells is dependent on the maturation and differentiation status of DCs. This status is affected by the cellular lineage of the DCs and by signals that the cells receive from the environment and from T cells. A specific subpopulation of DCs (dendritic cell type 2 [DC2]) induces the development of T helper 2 (Th2) responses. Unregulated Th2 responses induce and cause inflammation in allergy and asthma. If it would be possible to target DC2 cells for prophylactic or therapeutic measures, then it may be possible to change the T cell response to allergens on a long-term basis. In the past few years, there have been major research efforts to elucidate molecular determinants of DC maturation. This review summarizes the new findings and their potential for future clinical application. PMID- 15781124 TI - UDP-glucuronosyltransferases and clinical drug-drug interactions. AB - UDP-glucuronosyltransferase (UGT) enzymes catalyze the conjugation of various endogenous substances (e.g., bilirubin) and exogenous compounds (e.g., drugs). The human UGT superfamily is comprised of 2 families (UGT1 and UGT2) and 3 subfamilies (UGT1A, UGT2A, and UGT2B). Many of the individual UGT enzymes are expressed not only in liver but also in extrahepatic tissues, where the extent of glucuronidation can be substantial. Several others (e.g., UGT1A7, UGT1A8, and UGT1A10) are expressed only in extrahepatic tissues. The molecular regulation of UGT enzyme is still not fully understood, but various transcription factors appear to play a regulatory role. The expression of individual UGT enzymes is subject to genetic polymorphism and these enzymes can be inhibited or induced by xenobiotics. Experimental evidence in humans indicates that the glucuronidation of acetaminophen, codeine, zidovudine, carbamazepine, lorazepam, and propafenone can influenced by specific interacting drugs. In contrast, the glucuronidation of diflunisal, morphine, naproxen, and temazepam is not affected appreciably by the drugs investigated to date. In general, UGT-mediated human drug interaction studies are difficult to interpret. The factors that complicate the interpretation of this type of drug interaction data are discussed. PMID- 15781125 TI - The efficacy of cidofovir treatment of mice infected with ectromelia (mousepox) virus encoding interleukin-4. AB - Improved vaccines and therapies for virulent poxvirus infection are required, particularly in the light of recent threats of bioterrorism. Cidofovir (HPMPC) is an acyclic nucleoside analog with proven efficacy against poxviruses. Here, we evaluated HPMPC in mice given a recombinant ectromelia virus (ECTV) encoding interleukin-4 (ECTV-IL-4) that is highly immune suppressive. Mousepox-sensitive BALB/c mice given HPMPC for five consecutive days after infection were protected against the lethal effects of a control ECTV recombinant, although they suffered a chronic form of mousepox disease. High doses of the drug resulted in a milder localized disease. In contrast, HPMPC failed to protect mousepox-resistant C57BL/6 mice against ECTV-IL-4, although its lethal effects were delayed by five daily doses of 20 mg/kg or a single dose of 100 mg/kg. Higher daily doses further delayed mortality, although the majority of animals eventually succumbed to infection. It appears that HPMPC inhibited ECTV-IL-4 replication without clearance, with the virus having a lethal effect when the drug was removed. Resistance of ECTV-IL-4 to HPMPC treatment may relate to the virus's ability to inhibit antiviral cell-mediated immunity. Interestingly, ECTV-IL-4-mediated immune suppression was not accompanied by a reduction in systemic IFN-gamma expression, suggestive of an alternative or highly localized suppressive mechanism. PMID- 15781126 TI - Cranberry juice constituents affect influenza virus adhesion and infectivity. AB - Cranberry juice contains high molecular weight materials (NDM) that inhibit bacterial adhesion to host cells as well as the co-aggregation of many oral bacteria. Because of its broad-spectrum activity, we investigated NDM's potential for inhibiting influenza virus adhesion to cells, and subsequent infectivity. Hemagglutination (HA) of red blood cells (RBC) caused by representatives of both influenza virus A subtypes (H1N1)and H3N2) and the B type was inhibited by NDM at concentrations of 125 microg/ml or lower, which is at least 20-fold lower than that usually found in cranberry juice. A dose-response effect of NDM on HA was demonstrated. The infectivity of the A and B types was significantly reduced by preincubation with NDM (250 microg/ml), as reflected by the lack of cytopathic effect on Madine-Darby canine kidney (MDCK) cells and the lack of HA activity in the media of infected cells. The effect of NDM was also tested after A or B type viruses were allowed to adsorb to and penetrate the cells. Various levels of reduction in virus tissue culture infective dose TCID50 were observed. The effect was most pronounced when NDM was added several times to the infected MDCK cells. Our cumulative findings indicate that the inhibitory effect of NDM on influenza virus adhesion and infectivity may have a therapeutic potential. PMID- 15781127 TI - Anti-gene peptide nucleic acid targeted to proviral HIV-1 DNA inhibits in vitro HIV-1 replication. AB - Highly active antiretroviral therapy (HAART) is unlikely to affect reservoirs of HIV in latently infected cells. Anti-gene compounds, such as peptide nucleic acids (PNAs), which block transcriptional activity via sequence-specific invasion of double-stranded DNA may be an effective strategy to target cells harbouring proviral HIV DNA. Here we show that a PNA oligomer (PNA(HIV)), 15 bases in length, linked to a nuclear localization signal (NLS), substantially suppressed HIV-1 replication in chronically infected lymphocytes and macrophages and efficiently prevented mitogen-induced HIV-1 reactivation in lymphocytes, as determined by HIV-p24 antigen production in supernatants and FACS analysis for intracellular HIV accumulation. In contrast, a mismatched PNA did not show any effect on HIV expression. Semi-quantitative RT-PCR and quantitative real-time RT PCR demonstrated a decrease of HIV RNA expression in infected cells treated by PNA(HIV) indicating that inhibition of HIV-1 replication occurred at the transcription step. In conclusion, the use of anti-gene PNA to target the HIV-1 proviral DNA in the quest for new antiretroviral agents appears quite promising. PMID- 15781128 TI - Effects of IL-12 and IL-18 on HBcAg-specific cytokine production by CD4 T lymphocytes of children with chronic hepatitis B infection. AB - The influence of IL-12 and IL-18 was evaluated on hepatitis B core antigen (HBcAg)-specific cytokine production (IFN-gamma, IL-4, IL-5 and IL-10) by CD4 T lymphocytes isolated from peripheral blood of children with chronic hepatitis B. CD4 T cells were isolated from peripheral blood of 20 children with chronic active hepatitis B, cultured for 48h in presence of rHBcAg and of co-stimulators, IL-12 or IL-18 or IL-12+IL-18 or in their absence (control). Production of studied cytokines was examined using the ELISPOT assay. Co-stimulation with IL-12 or IL-18 was found to significantly augment the HBcAg-specific secretion of IFN gamma. However, the most pronounced stimulatory effect was observed in the presence of IL-12+IL-18 and resulted in peak levels of IFN-gamma production. The obtained results allowed concluding that the anti-HBV activity of Th1 lymphocytes is strongly induced by IL-12+IL-18 and may contribute to viral clearance in children with chronic hepatitis B infection. PMID- 15781129 TI - Ethanol extract of Polygonum cuspidatum inhibits hepatitis B virus in a stable HBV-producing cell line. AB - Chronic hepatitis B virus (HBV) infection is endemic in Asia and its consequences are among the major public health problems in the world. Unfortunately, the therapeutic efficacies of present strategies are still unsatisfactory with a major concern about viral mutation. In search of effective antiviral agent, we examined the efficacy of extracts of Polygonum cuspidatum Sieb. et Zucc. (P. cuspidatum) against HBV in HepG2 2.2.15 cells by quantitative real time polymerase chain reaction. The expressions of viral antigens, HBeAg and HBsAg, were also determined by enzyme linked immunosorbent assay. The ethanol extract of P. cuspidatum could inhibit dose-dependently the production of HBV (p<0.0001) with an effective minimal dosage of 10 microg/ml. The water extract of P. cuspidatum might also inhibit the production of HBV at a higher dosage. The expression of HBsAg was significantly increased by both ethanol extract and water extract of P. cuspidatum dose-dependently (p<0.0001) and time-dependently (p<0.0001). Higher dose of water extract of P. cuspidatum (30 microg/ml) could inhibit the expression of HBeAg (p<0.05). The extract of P. cuspidatum might contain compounds that would contribute to the control of HBV infection in the future. However, its promoting effect on the expression of HBsAg and its cytotoxicity should be monitored. Further purification of the active compounds, identification and modification of their structures to improve the efficacy and decrease the cytotoxicity are required. PMID- 15781131 TI - Differential profile of genes expressed in hemocytes of White Spot Syndrome Virus resistant shrimp (Penaeus japonicus) by combining suppression subtractive hybridization and differential hybridization. AB - White Spot Syndrome Virus (WSSV) is the major viral pathogen of culture shrimp. Although remarkable progress has been made in characterizing the WSSV genome, information concerning the antiviral process of host is still limited. To identify the genes differentially expressed along with their expression profile in the hemocytes of the virus-resistant shrimp, suppression subtractive hybridization (SSH) and differential hybridization (DH) were employed. Relying on the sequences identified in the subtractive cDNA library, 30 genes were characterized to be involved in the antiviral process as defense-relevant, among them, 22 are found for the first time in penaeid shrimp. The most interesting finding is that the interferon-like protein (IntlP) and (2'-5') oligo(A) synthetase-like protein (data not shown) known as the antiviral factors showed increased expression in virus-resistant shrimp and the non-specific antiviral activity of IntlP protein was verified by cytotoxicity experiment. A number of proteins with certain similarities to the components of the complement and cytokines system in vertebrates were also found in the subtracted library. The high expression of redox-related factors (NADH dehydrogenase, glutathione peroxidase and transcription factor AP-1 precursor), plasma defensive protein (C type lectin and laminin-like protein) and translationally controlled tumor protein (TCTP) in the virus-resistant shrimp suggested that they are essential components participating in the antiviral process. Our work provides a wide array of genes differentially expressed in the virus-resistant shrimp, and a framework for further studies aimed at antiviral mechanism in shrimp. PMID- 15781130 TI - Antiviral activity of serum from the American alligator (Alligator mississippiensis). AB - Serum from wild alligators was collected and tested for antibiotic activity against three enveloped viruses using cell-based assays. Alligator serum demonstrated antiviral activities against human immunodeficiency virus type 1 (HIV-1; IC50=0.9%), West Nile virus (WNV; IC50=4.3%), and Herpes simplex virus type 1 (HSV-1; IC50=3.4%). The inhibitory concentration (IC50) is defined as the concentration of serum that inhibits 50% of viral activity. The antiviral effects of the alligator serum were difficult to evaluate at high concentrations due to the inherent toxicity to the mammalian cells used to assay viral activities. The TC50 (serum concentration that reduces cell viability to 50%) values for the serum in the HIV-1, WNV, and HSV-1 assays were 32.8, 36.3 and 39.1%, respectively. Heat-treated serum (56 degrees C, 30 min) displayed IC50 values of >50, 9.8 and 14.9% for HIV-1, WNV and HSV-1 viruses, respectively. In addition, the TC50 values using heat-treated serum were substantially elevated for all three assays, relative to untreated serum (47.3 to >50%). Alligator serum complement activity has been shown to be heat labile under these conditions. HIV 1 antiviral action was heat-sensitive, and thus possibly due to the action of serum complement, while the anti-WNV and anti-HSV-1 activities were not heat labile and thus probably not complement mediated. PMID- 15781132 TI - Involvement of HIV-1 protease in virus-induced cell killing. AB - Acute infection of human CD4+ cells with cytopathic strains of HIV-1 causes rapid cell death. The role played by HIV-1 protease (PR) in virus-induced cell killing was investigated by subjecting C8166 cells to a single round of infection. The presence of HIV-1 PR inhibitor saquinavir from 24h post-infection prevented virus induced cell lysis. This inhibitor caused only a small reduction in the number of infected cells and in the expression of HIV-1-specific proteins. Moreover, treatments that block HIV-1 reinfection, such as AZT or the anti-CD4 antibody leu3.a, exerted little effect on virus-induced cell death. Thus, the specific inhibition of HIV-1 protease reduced the extent of both necrosis and apoptosis in C8166 cells such that most cells survived HIV-1 infection. Continued treatment of the infected cells with saquinavir led to the progressive suppression of HIV-1 expression; no viral proteins being detected 10 days after primary infection. Notably, reactivation of HIV-1 protease in these cells by removing the saquinavir triggered virus replication and cell lysis. These findings may contribute towards a better understanding of HIV-1 pathogenesis, and emphasise the potential of the virus protease as a key therapeutic target in AIDS treatment. PMID- 15781133 TI - Development of a cotton rat-human metapneumovirus (hMPV) model for identifying and evaluating potential hMPV antivirals and vaccines. AB - Hispid cotton rats were inoculated with two different human metapneumovirus (hMPV) subtype A strains and one subtype B hMPV. Although no overt disease was seen in any virus-inoculated animal, following an eclipse phase, significant pulmonary virus titers were observed in every hMPV-inoculated animal through day 7 post virus inoculation (p.i.) and in most through day 10. Peak virus titers occurred four days p.i., while virus-induced histopathology was most evident in lung sections obtained from animals 7 to 10 days p.i. The latter consisted primarily of desquamating and hypertrophic columnar epithelial cells lining the bronchi and bronchioles and the presence of large numbers of leukocytes in and around the bronchi and bronchioles. In fluorescent antibody studies, virus antigen-specific fluorescence was most evident in the desquamating tall columnar epithelial cells lining bronchi and bronchioles, in pneumocytes lining alveoli and in single or small groups of free cells, most probably leukocytes, present in the lumen of alveoli, bronchi and bronchioles. Virus was generally not detected in inoculated animals >10 days p.i. Although the pattern of virus replication in cotton rats was similar for all the three virus stains, the B subtype consistently grew to lower levels than the two A strains. Regardless, these findings indicate that hMPV replicates in cotton rats and that these animals may be used as a small animal model of hMPV infection and to facilitate the identification and development of vaccines and antivirals for preventing and/or ameliorating infections caused by this virus. PMID- 15781134 TI - The fd phage and a peptide derived from its p8 coat protein interact with the HIV 1 Tat-NLS and inhibit its biological functions. AB - Filamentous fd bacteriophages are used to construct phage-display peptide libraries, which have been instrumental in selecting peptides that interact with specific domains within target molecules. Here we demonstrate that the fd bacteriophage itself, as well as NTP8 - a synthetic peptide derived from it and bearing amino acids 1-20 of the phage p8 protein - interact with the nuclear localization signal (NLS) of the HIV-1 Tat protein. Accordingly, fd bacteriophage and the NTP8 peptide inhibit binding mediated by the Tat-NLS to the nuclear import receptor importin beta and Tat-NLS-mediated translocation into cell nuclei. The NTP8 peptide, at 100 microM concentration, also caused about 50% inhibition of HIV-1 propagation in cultured cells. The fd bacteriophage prevents heparan sulfate proteoglycans-mediated uptake of extracellular Tat by target cells and consequently transactivation of a chloramphenicol acetyltransferase (CAT) reporter gene. A BSA-NTP8 conjugate inhibits Tat-NLS-mediated binding to heparin immobilized on a BIAcore surface. BLAST analysis of the NTP8 amino-acid sequence revealed similarity to sequences in several human proteins, including ADA2 and CD53. PMID- 15781135 TI - Restoration materials for primary molars-results from a European survey. AB - OBJECTIVE: The aim of this study was to obtain an overview of the materials and restorative techniques taught for Class I and Class II restorations in primary molars of different European departments for paediatric dentistry and to compare the results to those for North America. METHODS: The forms were sent to the chairmen of the departments of paediatric dentistry or-when included into the department for operative dentistry-to the director of 200 universities in 32 European Countries. The questions concerned the preferred materials and techniques, as well as the indications or contraindications for the use of the different tooth coloured materials. RESULTS: Forty-three percent of the universities from western European countries (excluding Germany) responded to the survey. A 97% answer rate was obtained from German Schools. Of the other European regions, the response rate varied remarkably (24-54%). Also, opinions on materials used and methods applied were different from country to country. Only in a few regions (Eastern Europe), amalgam remains the filling material of first choice for Class I and II cavities in primary molars, whereas in others, the use has been restricted and tooth coloured materials, especially compomers or resin modified glass ionomer cements, are being preferred. CONCLUSIONS: Great diversity in teaching was observed not only among the European countries, but also within the countries themselves which makes it difficult to compare the results of our study to the ones obtained from the US. PMID- 15781136 TI - Influence of Carisolv on resin adhesion for two different adhesive systems to sound human primary dentin and young permanent dentin. AB - OBJECTIVE: This study evaluated the influence of Carisolv (Medi Team) for resin adhesion to sound human primary and young permanent dentin. METHODS: The buccal surfaces of 28 primary molars and 64 premolars were used. Two adhesive systems and resin composites were used; SE: Clearfil SE (Kuraray) and Clearfil APX (Kuraray), and SB: Single Bond (3M) and Z250 (3M). Six groups were prepared. Groups 1-2 were primary dentin and Groups 3-6 were permanent dentin. Groups 1 and 3: SE was used. Groups 2 and 4: treated with Carisolv and then primed, SE was used. Group 5: SB was used. Group 6: treated with Carisolv and then etched, SB was used. The microstructural effects of primer or etchant, and Carisolv plus primer or etchant applied to dentin were evaluated by SEM. In addition, the microstructure of the resin-dentin interfaces of each group was studied using SEM. Shear bond strengths (SBS) were tested, and the failed surfaces were observed using SEM. Data was statistically analyzed using ANOVA with subsequent application of Fisher's PLSD at p<0.05. RESULTS: The mean SBS (unit: MPa) of Groups 1-6 were: 27.8, 19.2, 21.3, 21.7, 6.7 and 7.6. The SBS of Group 2 was significantly lower than that of Group 1. There was no significant difference of the SBS among Groups 1 and 4, 2, 3 and 4, and 5 and 6. In SE groups, the hybrid layer for primary dentin was thicker than that for permanent dentin. CONCLUSIONS: Carisolv treatment before priming significantly decreased the SBS to primary dentin in SE groups, but did not influence the SBS to permanent dentin in both SE and SB groups. PMID- 15781137 TI - Detection of bacteraemias during non-surgicalroot canal treatment. AB - Some dental procedures initiate a bacteraemia. In certain compromised patients, this bacteraemia may lead to distant site infections, most notably infective endocarditis. OBJECTIVE: To investigate whether a detectable bacteraemia was produced during non-surgical root canal therapy. METHODS: Thirty patients receiving non-surgical root canal therapy were studied. Three blood samples were taken per patient: pre-operatively, peri-operatively and post-operatively. In addition, a paper point sample was collected from the root canal. The blood samples were cultured by pour plate and blood bottle methods. The isolated organisms were identified by standard techniques. Blood samples were analysed for the presence of bacterial DNA by the polymerase chain reaction (PCR). In two cases where the same species of organism was identified in the root canal and the bloodstream, the isolates were typed by pulsed field gel electrophoresis (PFGE). RESULTS: By conventional culturing, a detectable bacteraemia was present in 9 (30%) of the 30 patients who had no positive pre-operative control blood sample. In 7 (23.3%) patients, the same species of organism was identified in both the bloodstream and in the paper point sample from the root canal system. Overall, PCR gave lower detection rates compared with conventional culture, with 10 of 90 (11%) of the blood samples displaying bacterial DNA. PFGE typing was undertaken for two pairs of culture isolates from blood and paper points; these were found to be genetically identical. CONCLUSIONS: Non-surgical root canal treatment may invoke a detectable bacteraemia. PMID- 15781138 TI - Survival of glass fibre reinforced composite post restorations after 2 years-an observational clinical study. AB - OBJECTIVES: The aim of this study is to provide prospective clinical data for the survival of postendodontic reconstructions of teeth with varying degrees of hard tissue loss using tapered or parallel-sided post shapes. METHODS: Eighty-three patients got 105 glass fibre reinforced posts of tapered (Luscent Anchors, Dentatus, Sweden) and parallel-sided, serrated (FibreKor, Jeneric Pentron, USA) post shape. A dual curing hybrid composite Compolute (3M ESPE, Germany) was used as luting material, EBS-Multi (3M ESPE) as adhesive system and Clearfil Core (Kuraray, Japan) for core built-up. The restorations were followed for a minimum of 24 months. The statistical analysis was performed on a random sub-sample of one restoration per subject. The Fisher exact test was used to compare frequencies of failures after 12 and 24 month. A Kaplan-Meier-analysis was used to analyse time-to-failure in both groups. Differences of survival time between post types were tested with the log-rank test. RESULTS: 3.8% of the restorations failed after 12 month, 12.8% after 24 month, respectively. The main failure type observed was post fractures. All but one failed teeth could be restored. There was no difference in failure frequency between post types after 12 or 24 months. The log-rank test showed no differences in survival between the two types of post (p=0.37). CONCLUSION: Parallel-sided and tapered glass fibre posts result after 2 years of clinical service in an equal rate of survival. PMID- 15781139 TI - In vitro studies on the effect of sodium tripolyphosphate on the interactions of stain and salivary protein with hydroxyapatite. AB - OBJECTIVES: To study properties of sodium tripolyphosphate (STP) relevant to inhibition or removal of dental stain in vitro. METHODS: The effects of STP and other phosphates on adsorption of a dietary chromogen (black tea polyphenol) and salivary protein to hydroxyapatite (HA) powder were studied by analysing loss of protein or tea stain from solutions mixed with HA or HA pre-treated with the test agents. The effects on desorption of protein and stain from HA were studied by analysis of water or solutions of test agents mixed with HA or HA pre-treated with saliva or tea solution. RESULTS: At concentrations and pH representative of those likely to occur in the mouth, STP inhibited adsorption of salivary protein and black tea polyphenol to, and desorbed these substances from, HA surfaces. Adsorption and desorption of protein and stain were not influenced by pH of the STP solutions but adsorption varied with concentration. STP showed equivalent effectiveness with respect to salivary protein adsorption and desorption as a longer-chain condensed phosphate. The inhibitory activity of HA-bound STP on adsorption of salivary protein and stain resisted extensive washing. CONCLUSIONS: STP is likely to be an effective agent for inhibiting and removing dental stain, whether bound directly to mineralised surfaces or indirectly via salivary pellicle. PMID- 15781140 TI - Three-dimensional evaluation of gap formation of cervical restorations. AB - OBJECTIVES: In some studies gap formation has been evaluated in just one section of the restorative. This in vitro study aimed to design a quantitative three dimensional method for evaluation of the contraction gap in restoratives. METHODS: Cervical cavities were prepared on buccal, palatal or lingual surfaces in human extracted molars and were then filled with resin composites. Specimens were reduced every 100 microm in a direction parallel to the tooth axis, and perpendicular to the cavity floor from one proximal side to the other. The sequence of reducing the sections by 100 microm, image taking (250x) and observation of these images (maximal 2500x) were repeated. Three-dimensional images of the contraction gap were made using analytical software and the proportions of the interface with gap formation calculated. RESULTS: The mean proportions of the interface with gap formation of the self-etching system (Clearfil Liner Bond II Sigma) was 41.7+/-6.3% and that of the self-priming system (Single Bond) was 38.2+/-3.9%; there was no significant difference. CONCLUSIONS: Approximate three-dimensional images of the in vitro contraction gap could be drawn and the mean proportions of the interface with gap formation could be more precisely calculated than by previous methods. PMID- 15781141 TI - An in vitro investigation of microtensile bond strengths of two dentine adhesives to caries-affected dentine. AB - OBJECTIVES: To compare the micro-tensile bond strengths of two different adhesive systems (ABF (Clearfil Protect Bond), Kuraray Medical Inc., Tokyo, Japan) and Prime & Bond NT (PBNT, Dentsply, Konstanz, Germany) bonded to caries-affected dentine retained after chemo-mechanical caries removal using Carisolv gel, with that retained after excavation using conventional hand instrumentation. METHOD: Twenty, adult, human extracted carious teeth were used in this split tooth study with bur-cut cavities in sound dentine acting as controls. After clinical caries excavation, the occlusal cavities in each experimental group were restored with either bonding system plus composite. Matchstick-shaped samples through the bond interfaces were sectioned and microtensile bond strengths recorded. Scanning electron microscopy (SEM) was used to ascertain the mode of failure at the restoration-dentine interface. RESULTS: Statistical analysis of the bond strength data showed that for the ABF group, there was no difference in bond strengths between the controls and Carisolv group but these values were significantly higher than those for the hand-excavated samples. PBNT samples showed no significant differences in any of the three test groups, with wider ranging data sets. SEM analysis indicated a variety of failures at the interface including cohesive failures within the caries-affected dentine itself. CONCLUSIONS: From the data generated by this study it could be concluded that microtensile bond strengths of PBNT/composite restorations to caries-affected dentine in clinical cavities were statistically comparable to those to sound dentine. In the ABF/composite restored group (self-etched), the use of conventional hand excavation appeared to weaken the bond strength to the remaining caries-affected dentine. However, the use of Carisolv gel excavation did not compromise bond strengths to caries-affected dentine in either group tested. PMID- 15781142 TI - A methodology for testing the erosive potential of sports drinks. AB - OBJECTIVES: The aim of the study was to develop and test a methodology in situ, which simulated the consumption of sports drinks. A secondary aim was to assess the acceptability of the method to sedentary participants. METHODS: To select the sports drink for the study in situ, five commercially available sports drinks were examined for erosive potential in vitro. The study in situ was a single centre, 2-period, 2-treatment crossover study to compare the erosive effect of a commercially available sports drink (Test), with that of mineral water (Control), over 10 day periods on 10 healthy volunteers. Subjects wore upper removable appliances containing two human enamel specimens from 9 a.m. to 5 p.m. The regimen of intake of the drinks was 350 ml in 10, 5-min rest, 650 ml in 25, 5-min rest, 500 ml in 10 and 5-min rest. Measurements of enamel loss were made on samples after 5 and 10 days by profilometry. RESULTS: The in situ study showed a statistically significant difference in erosive potential between the test and control beverages. No specimen exposed to the control beverage displayed appreciable erosion. Erosion occurred with the test drink, but to a variable degree between subjects. The subjects unanimously found the drinking regimen unpleasant. CONCLUSIONS: The sports drink caused significantly more erosion in situ than water and as seen in other studies, there was marked variation in susceptibility to erosion between subjects. The new drinking regimen was designed to simulate pre, during and post-exercise intake. Although all the sedentary subjects participating in this study reported that they found the volume of fluids consumed over a short period of time excessive it is unlikely that this would prove problematic in the exercise environment. PMID- 15781143 TI - Microhardness of acid-treated and resin infiltrated human dentine. AB - OBJECTIVE: The aim is to determine if superficial or deep dentine microhardness (MH) is affected by different chemical dentine pre-treatments performed for resin bonding. METHODS: Dentine discs of superficial (SD) and deep dentine (DD) were obtained by transversally sectioning the crowns of human third molars. Knoop MH was measured after different treatments: (1) polished up to 4000 grit, (2) polished and etched (37% ortophosphoric acid for 15 s), (3) resin (Single Bond SB-) infiltrated dentine after acid etching, (4) polished, etched and treated with 5% NaOCl for 2 min, (5) resin infiltrated (SB) after etching and NaOCl treatment. For resin infiltrated surfaces care was taken in order to remove the excess adhesive layer, and obtain infiltrated dentine, by measuring thickness of the sample before resin infiltration and polishing after resin infiltration until obtaining the initial thickness of the sample. Indentations (n=20) were performed on moist surfaces with a standard Knoop MH tester (20 g, 5 s). Data were expressed in Knoop Hardness Numbers (KHN) and analysed by ANOVA and multiple comparisons (P<0.05). RESULTS: Dentine KHN decreased on both SD and DD after acid etching. NaOCl treatment after acid etching did not affect MH on SD, but KHN of DD was lowered. Resin infiltration increased KHN but did not recover the initial MH values in both SD and DD. CONCLUSIONS: Treating dentine with either H3PO4 or NaOCl caused marked reduction of its surface hardness and subsequent resin infiltration was not capable to restore it. PMID- 15781144 TI - Expression of human neuronal protein 22, a novel cytoskeleton-associated protein, was decreased in the anterior cingulate cortex of schizophrenia. AB - Human neuronal protein 22 (hNP22) is a novel neuron-specific protein featuring numerous motifs previously described in cytoskeleton-associating and signaling proteins. Because previous studies have supported abnormalities in neuronal cytoarchitecture and/or development in the schizophrenia brain, we examined the expression of hNP22 in the anterior cingulate cortex, the hippocampus and the prefrontal cortex of schizophrenic and normal control postmortem brains using high-sensitive immunohistochemistry. Seven schizophrenic and seven age- and sex matched control brains were examined. The ratio of hNP22-immunopositive cells/total cells was significantly reduced in layer V (p=.020) and layer VI (p=.022) of the anterior cingulate cortex of schizophrenic brain compared with controls. In contrast, there were no significant changes observed in the hippocampus and the prefrontal cortex. These results suggest that altered expression of hNP22 may be associated with modifications in neuronal cytoarchitecture leading to dysregulation of neural signal transduction in the anterior cingulate cortex of the schizophrenia brain. PMID- 15781146 TI - Postural control during quiet standing following cervical muscular fatigue: effects of changes in sensory inputs. AB - The purpose of the present experiment was to investigate the effects of cervical muscular fatigue on postural control during quiet standing under different conditions of reliability and/or availability of somatosensory inputs from the plantar soles and the ankles and visual information. To this aim, 14 young healthy adults were asked to sway as little as possible in three sensory conditions (No vision, No vision-Foam support and Vision) executed in two conditions of No fatigue and Fatigue of the scapula elevator muscles. Centre of foot pressure (CoP) displacements were recorded using a force platform. Results showed that (1) the cervical muscular fatigue yielded increased CoP displacements in the absence of vision, (2) this effect was more accentuated when somatosensation was degraded by standing on a foam surface and (3) the availability of vision allowed the individuals to suppress this destabilising effect. On the whole, these findings not only stress the importance of intact cervical neuromuscular function on postural control during quiet standing, but also suggest a reweigthing of sensory cues in balance control following cervical muscular fatigue by increasing the reliance on the somatosensory inputs from the plantar soles and the ankles and visual information. PMID- 15781145 TI - Neuropharmacological profiles of antagonists of group II metabotropic glutamate receptors. AB - Glutamatergic abnormalities play roles in several psychiatric disorders. Glutamate acts at two classes of receptors, ionotropic and metabotropic glutamate receptors (mGluR), the latter is classified into three group, based on receptor homology and signaling mechanisms. Among them, recent pharmacological and histochemical studies suggest that the group II mGluR (mGluR2 and mGluR3) plays crucial roles in the control of emotional states. We previously reported that MGS0039, a selective group II mGluR antagonist, exhibited dose-dependent antidepressant-like effects in some animal models. However, the mechanism by which group II mGluR antagonists exhibit such effects is still unclear. In the present two studies, we examined neuropharmacological effects of group II mGluR antagonists on monoaminergic neurons. In an electrophysiological study, MGS0039 dose-dependently and significantly increased the firing rate of dorsal raphe nucleus (DRN) serotonergic neurons. LY341495, another group II mGluR antagonist, also increased DRN serotonergic neural activity significantly. Consistent with the findings of this electrophysiological study, MGS0039 significantly increased extracellular level of serotonin in rat medial prefrontal cortex in a microdialysis study. In contrast, MGS0039 had no effect on the activity of locus coeruleus noradrenergic neurons. These findings suggest that modulation of serotonergic neuron might be, at least in part, responsible for the antidepressant-like effects of group II mGluR antagonists. PMID- 15781147 TI - Molecular dissection of tropisetron, an alpha7 nicotinic acetylcholine receptor selective partial agonist. AB - The alpha7 nicotinic acetylcholine receptor (nAChR)-selective partial agonist tropisetron is a conjugate of an indole and a tropane group. We tested compounds structurally related to either the indole or tropane domains of tropisetron on oocytes expressing human alpha7. alpha4beta2, or alpha3beta4 nAChR or rat 5HT(3A) receptors. The simple compounds tropane and tropinone had alpha7-selective agonist activity comparable to that of tropisetron. Tropinone was more efficacious than tropisetron but 100-fold less potent. Some tropane compounds had antagonist activity on alpha3beta4 nAChR but no effect on alpha4beta2 nAChR. Some tropanes also affected the responses of 5HT3 receptors to serotonin. Tropisetron was more potent at inhibiting alpha3beta4 receptors (IC(50)=1.8+/-0.6) than was tropane or tropinone, suggesting that the presence of the indole group has a large impact on the potency of tropisetron, both as an alpha7 agonist and as an alpha3beta4 antagonist. The further reduced structures of dimethyl piperidinium and 1-methylpyrrolidine also had agonist activity on alpha7 receptors, suggesting that the minimal activating pharmacophore of these compounds, as with tetramethylammonium, may simply be the charged nitrogen, while additional structure elements impact subtype selectivity, potency, and efficacy. It has previously been reported that 5-hydroxyindole (5HI) can potentiate alpha7 receptor responses to acetylcholine (ACh). However, the site where 5HI binds to the receptor is not known. We tested the hypothesis that the tropisetron binding site might overlap the 5HI site and thereby produce a block of 5HI potentiation. Our results indicate that the indole portion of tropisetron is not likely to be binding to the same site where 5HI binds to potentiate alpha7 receptor responses since 5HI can greatly potentiate responses of tropisetron, tropinone, and other partial agonists such as 4OH-GTS-21. PMID- 15781148 TI - Auditory evoked potentials to tones and syllables in adults: evidence of specific influence on N250 wave. AB - Late auditory evoked potentials were recorded in eight adults according to stimulus duration (50ms versus 250ms) and to speech nature of the stimulus (tones versus syllables). The main effect of these parameters concerned the negative fronto-central wave, which occurs beyond 200ms termed N250 in this paper; it had a greater amplitude and a longer latency when tone duration increased from 50 to 250ms and a longer latency, a greater amplitude and a longer time-course in response to syllables compared to tones. Moreover, this wave was inversed in polarity at the mastoid sites, indicating the involvement of generators of the N250 wave in the supratemporal auditory cortex. These results showing the sensitivity of the N250 wave to tone duration and to speech stimuli indicated that this electrophysiological index might allow further assessment of cortical activity involved in speech stimuli processing. PMID- 15781149 TI - TrkB-deficient mice show diminished phase shifts of the circadian activity rhythm in response to light. AB - Brain-derived neurotrophic factor (BDNF) has been implicated in the mechanism underlying the circadian sensitivity of the clock in the hypothalamic suprachiasmatic nucleus (SCN) to the phase-shifting effects of light. In the present study, we examined the role of the cognate receptor for BDNF, the TrkB tyrosine kinase, in the photic regulation of the SCN clock by determining whether the phase-shifting action of light is impaired in mice with targeted mutation of the TrkB gene. In comparison with wild-type littermates, heterozygous TrkB mutant mice (trkB(+/-)) showed marked reductions in SCN and cortical levels of this neurotrophin receptor that were accompanied by decreases in the amplitude of light-induced phase shifts during the subjective night. These results provide further evidence indicating that BDNF-mediated signaling through the TrkB receptor is an important process in the gating of SCN responses to light and its phase-shifting effects on circadian rhythms. PMID- 15781150 TI - Beta rebound after different types of motor imagery in man. AB - We investigated the behavior of short-lasting beta bursts (beta rebound, beta ERS) induced after imagination of hand, foot or tongue movement. Nine able-bodied subjects were asked to imagine one type of movement following the presentation of a visual cue stimulus. EEG was recorded from 60 closely spaced electrodes placed over frontal, central and parietal areas. Spatiotemporal ERD/ERS patterns were evaluated in form of time-frequency maps. Seven out of nine subjects displayed a significant short-lasting beta power increase (beta rebound) after termination of imagery of movement of both feet in the frequency range of 23-29Hz with a clear maximum at the vertex. In contrast, only two subjects displayed a clear beta rebound at the vertex after imagery of one of the hands, although five exhibited a significant beta rebound at the contralateral side of the head. None showed a beta rebound after tongue motor imagery. The midcentrally located beta rebound is interpreted as an electrophysiological correlate of a simultaneous "resetting" of overlapping neural networks in the foot representation areas and the SMA. PMID- 15781151 TI - Effect of Muller cell co-culture on in vitro permeability of bovine retinal vascular endothelium in normoxic and hypoxic conditions. AB - Muller cell dysfunction may contribute to the early pathological changes associated with conditions such as diabetes, that cause breakdown of the blood retinal barrier. In this study we used an in vitro model of the blood-retinal barrier to investigate Muller cell effects on retinal vascular endothelial cell monolayer permeability under normoxic (20% oxygen) and hypoxic (1% oxygen) conditions. Second passage bovine retinal capillary endothelial cells were co cultured with retinal Muller cells on opposite sides of a 0.4 microm pore size polycarbonate Transwell filter or in medium that was continually conditioned by Muller cells. Permeability changes were observed for up to 24h of hypoxia by measurement of [(3)H]-inulin and [(14)C]-albumin flux across the endothelial cell monolayer. Endothelial cell barrier function was enhanced by co-culturing with Muller cells under normoxic conditions. Under hypoxic conditions however, the barrier was significantly impaired after 12h of co-culture with Muller cells. These results shed more light on the trophic effect of Muller cells on the blood retinal barrier, suggesting a critical role in the maintenance and regulation of the barrier in health and during disease. PMID- 15781152 TI - Extracellular ascorbate modulates cortically evoked glutamate dynamics in rat striatum. AB - To determine if extracellular ascorbate, which may increase by several hundred micromolar in striatum during behavioral activation, directly alters glutamate transmission, we monitored striatal glutamate transients evoked by electrical stimulation of cerebral cortex in anesthetized rats tested with varying concentrations of ascorbate (0, 50, 200, and 500 microM) by reverse dialysis. Capillary electrophoresis coupled with laser-induced fluorescence detection was used to analyze dialysates collected at 3-s intervals. Ascorbate elevated striatal glutamate in a concentration-dependent fashion. Addition of 500 microM ascorbate not only more than doubled basal glutamate levels relative to the ascorbate-free condition, but significantly increased both the magnitude of the electrically evoked glutamate response as well as its subsequent return to baseline. In fact, the time required to return to within 10% of the pre stimulation baseline increased by >100s. Reverse dialysis of iso-ascorbate, in contrast, had no effect on stimulation-evoked glutamate release arguing against an antioxidant effect. It appears, therefore, that the level of extracellular ascorbate plays a critical role in regulating corticostriatal glutamate transmission. PMID- 15781153 TI - BAG-1 associates with the polyglutamine-expanded huntingtin aggregates. AB - Huntington's disease (HD) is characterised by the proteolytic production of N terminal fragments of huntingtin containing polyglutamine repeats that forms intracellular ubiquitinated aggregates in the affected neurons. Using cellular and transgenic mice model of HD, we report here that BAG-1 co-immunoprecipitates with the polyglutamine-expanded truncated N-terminal huntingtin (tNhtt) and associates with their aggregates through its interaction with the chaperones Hsc70/Hsp70. We further demonstrate that the over expression of BAG-1 protects polyglutamine-expanded tNhtt induced cell death. Since, BAG-1 is essential for cell survival, its association with tNhtt aggregates might disrupt its normal function and thereby promote polyglutamine-expanded tNhtt-induced cell death. PMID- 15781154 TI - Spatial navigation in the Morris water maze: working and long lasting reference memories. AB - Spatial navigation development in the Morris water maze (MWM) paradigm was studied in 70-day-old male Long-Evans hooded rats. During 5 consecutive days, rats' training consisted of a daily block of 10 trials. Escape latency was measured in each trial. Probe testing was performed every day immediately before and after the daily block of trials. In addition, a final probe was performed on Day 6. During the first 3 days of training, the escape latency became progressively shorter, showing an asymptotic trend on Days 4 and 5. Probe trials administered at the end of the first acquisition sessions showed clear preference for the target quadrant but this information was not recalled at probe trials given 24h later. The memory trace retrieved after 24-h delay was formed only after 30 trials received over three sessions. The probe trial given at the end of an acquisition session tests the efficiency of the working memory whereas the 24h delayed probe trial reflects better-consolidated spatial information corresponding to long lasting reference memory. It can be noted that the progressive shortening of escape latencies does not express closely the evolution of the rat's long lasting (consolidated) reference memory. This memory can be satisfactorily measured only by probe testing performed at an adequate delay after training. These considerations may be of some interest when interpreting the rat's performance in the MWM. PMID- 15781156 TI - Epithelial barrier dysfunction: a unifying theme to explain the pathogenesis of multiple organ dysfunction at the cellular level. AB - The multiple organ dysfunction syndrome (MODS) is the most common cause of death among patients requiring care in an ICU. There is widespread agreement that MODS is the clinical manifestation of a dysregulated inflammatory response. This article, however, summarizes some tantalizing data to support the view that derangements in the formation or function of specialized structures in epithelial cells, tight junctions, may be a key factor leading to lung, liver, gut, and perhaps kidney dysfunction associated with such conditions as sepsis and acute lung injury syndrome that are caused by dysregulated inflammatory processes. PMID- 15781157 TI - The pathophysiology of acute renal failure. AB - This article summarizes the pathophysiology of acute renal failure from both experimental and clinical points of view. Prerenal acute renal failure is an appropriate physiologic response to renal hypoperfusion and can complicate any disease characterized by either true hypovolemia or a reduction in the effective circulating volume. In acute tubular necrosis, the abrupt fall in glomerular filtration rate is thought to be caused by interplay of hemodynamic and tubular abnormalities. The postischemic kidney also shows a dramatic capacity for recovery. The molecular mechanisms and the effect of growth factors are also summarized. PMID- 15781158 TI - Sepsis and the kidney. AB - Acute renal failure (ARF) secondary to sepsis is a highly prevalent diagnosis in the ICU setting and continues to be associated with a high rate of morbidity and mortality. The pathophysiology of sepsis-induced acute renal failure involves ischemic or toxic injury to the renal tubular epithelia, resulting in necrosis or apoptosis, and clinically is characterized as acute tubular necrosis. The management of sepsis-induced ARF includes both conventional intermittent hemodialysis and continuous renal replacement therapies. Experimental therapies to improve outcomes in sepsis-associated ARF include the provision of plasmapheresis and adsorption therapies, and the recent development and deployment of a renal tubule assist device. PMID- 15781159 TI - Defining, quantifying, and classifying acute renal failure. AB - The syndrome of acute renal failure (ARF) is a common complication of critical illness. Like every other syndrome in the intensive care unit, it requires a consensus definition to progress with a research agenda that is dedicated to preventing and treating it. A consensus definition has been proposed and is being validated. ARF also requires quantification of severity because severity of functional loss is likely to determine the way in which ARF affects outcome. The new consensus definition also offers a quantification of severity. Finally, classification according to pathogenesis would be desirable but remains elusive. Important steps are being taken toward improving the outcome of these patients. Critical care physicians need to understand and participate in this process. PMID- 15781160 TI - Epidemiology and natural history of acute renal failure in the ICU. AB - This article focuses on the incidence, risk factors, and mortality of acute renal failure (ARF) in critically ill patients. Accurate epidemiologic assessment of ARF is still a problem; as long as there is neither a uniquely accepted definition of ARF nor definitions for end points to measure, results will remain heterogeneous and hard to compare. Mortality of ARF has remained high throughout the last decades, despite further development of modern treatment modalities. This indicates that ARF is not just a matter of loss of organ function that can easily be replaced easily by extracorporeal therapies, but is a condition additionally accompanied by systemic consequences which significantly impact on prognosis. PMID- 15781161 TI - Physiologic consequences of acute renal failure on the critically ill. AB - Acute renal failure (ACF) causes increased levels of uremic retention products, acidosis, and volume overload. These factors contribute to a dysregulated inflammatory response and decreased immunity, and interfere with normal function of many organs. ACF may play an important role in the etiology of multiple organ dysfunction. PMID- 15781162 TI - Contrast-induced nephropathy. AB - Contrast-induced nephropathy (CIN) is a leading cause of in-hospital acute renal failure in critically ill patients who undergo radiographic procedures. Critical care patients are at particular risk, often because of baseline renal dysfunction, older age, and the presence of diabetes. In addition, there are superimposed risks, including volume depletion, sepsis, and use of nephrotoxic drugs. The rates of CIN (defined as an increase in serum creatinine by >25% or 0.5 mg/dL) can be predicted by using multivariate tools. Prevention measures include adequate hydration, use of N-acetylcysteine and iso-osmolar contrast, and for patients who are at the highest risk, prophylactic hemofiltration. PMID- 15781163 TI - Prevention of acute renal failure. AB - Acute renal failure (ARF) complicates the clinical course of many critically ill patients and significantly adds to their morbidity and mortality. Efforts to develop techniques to prevent ARF or to facilitate its resolution largely have been unsuccessful. Aggressive hydration, minimizing nephrotoxins, and maintenance of "adequate" mean arterial pressure remain the main nonpharmacologic strategies to prevent ARF in the ICU. Increasing insight into mechanisms that lead to ARF prompted investigators to evaluate the role of novel therapeutic agents in the prevention of ARF. Recent data suggest that N-acetylcysteine may reduce the incidence of ARF secondary to radio-contrast agents. PMID- 15781164 TI - Oliguria, volume overload, Na+ balance, and diuretics. AB - This article discusses the pathophysiology and treatment of common fluid and electrolyte disorders in the ICU. The presence of oliguria should alert the intensivist to identify the underlying cause rather than to resort reflexively to measures, such as diuretics or dopamine, to establish urine flow. Hypo- and hypernatremia, which are exceedingly commonly in the ICU setting, also are discussed using a pathophysiologic approach. PMID- 15781165 TI - Metabolic, electrolytes, and nutritional concerns in critical illness. AB - This article discusses metabolic, electrolyte, and nutritional concerns in critical illness. PMID- 15781166 TI - Determinants of plasma acid-base balance. AB - An advanced understanding of acid-base physiology is central to the practice of critical care medicine. Intensivists spend much of their time managing problems that are related to fluids, electrolytes, and blood pH. Recent advances in the understanding of acid-base physiology occurred as the result of the application of basic physical-chemical principles of aqueous solutions to blood plasma. This analysis revealed three independent variables that regulate pH in blood plasma: carbon dioxide, relative electrolyte concentrations, and total weak acid concentrations. All changes in blood pH, in health and in disease, occur through changes in these three variables. This article reviews the physical-chemical approach to acid-base balance and considers clinical implications for these findings. PMID- 15781167 TI - Renal replacement therapy I: indications and timing. AB - The indications for initiation of renal replacement therapy in acute renal failure are controversial. Although volume overload, metabolic acidosis, hyperkalemia and other electrolyte disturbances, and overt uremic manifestations are commonly accepted indications for renal replacement therapy, specific criteria for initiation of therapy based on these conditions are highly subjective. Progressive azotemia in the absence of overt uremia is another common indication for renal replacement therapy although there is no consensus on the degree of azotemia that warrants initiation of therapy. The clinical data regarding timing and initiation of renal support in patients who have acute renal failure are reviewed. Definitive resolution of the appropriate indications and timing for initiation of renal replacement therapy in acute renal failure requires prospective evaluation in a randomized clinical trial. PMID- 15781168 TI - Renal replacement II: dialysis dose. AB - Improved survival of critically ill acute renal failure patients can be correlated with therapy dose. The overall solute elimination can be measured by the product of clearance and time (Kt), which is usually normalized for the volume of distribution (V) of the solute as "Kt/V." Setting a Kt/V threshold of 1.4 can guide clinicians toward adequate treatment. This is a slightly higher prescription than the current value for chronic dialysis. However, the true uraemic toxins probably diffuse among body compartments less readily than urea and, as such, the frequency of renal replacement therapy should be more important to its efficiency, and should be optimal with continuous therapy. In the absence of an optimal dialysis dose, it can only be recommended that the prescription should exceed that calculated to be "adequate." PMID- 15781169 TI - Renal replacement therapy III: IHD, CRRT, SLED. AB - Acute renal failure in critically ill patients is a growing clinical problem. Options for renal replacement therapy in these patients use convective and diffusive clearance and may be intermittent, as in classic hemodialysis, or continuous. New ways of delivering dialysis in the intensive care unit, such as sustained low-efficiency dialysis, are also under development. It may be that renal replacement therapy needs to be tailored to the needs of each individual patient. Current and future research studies should provide the answers to many of these questions. PMID- 15781170 TI - The future of renal support: high-flux dialysis to bioartificial kidneys. AB - Renal failure continues to cause a major burden of morbidity and mortality in both its acute and chronic forms, regardless of advances in current renal replacement therapies. A bioartificial kidney that includes a conventional dialysis filter and a renal tubule assist device containing approximately 10(8) renal proximal tubule cells was recently successfully engineered. This therapeutic modality may decrease the survival gap between current renal replacement therapies and healthy kidney functions and may lessen the heavy burden of morbidity and mortality associated with renal failure, both acute and chronic, in the near future. PMID- 15781171 TI - Increasing prescriber awareness of drug interactions in palliative care. PMID- 15781172 TI - Midazolam for long-term treatment of intractable hiccup. PMID- 15781173 TI - A multicenter study of the revised Edmonton Staging System for classifying cancer pain in advanced cancer patients. AB - The comparative analysis of analgesic interventions for cancer pain is greatly compromised by the lack of well-validated and clinically acceptable tools, which allow a composite classification of pain and patient population characteristics. Although the Edmonton Staging System (ESS) for cancer pain was developed for this purpose, clinical and research utility has been limited due to problems associated with the assessment of some items, especially in relation to definitions and terminology. To overcome these limitations, we designed a revised ESS (rESS) and conducted a multicenter study to determine its inter-rater reliability and predictive value. In revising the rESS, we hypothesized that patients with less problematic pain features would require a shorter time to achieve stable pain control, require less complicated analgesic regimens, be more responsive to opioid therapy, and use lower opioid doses. The rESS items include mechanism of pain, presence or absence of incidental pain, presence or absence of psychological distress and addictive behavior, and level of cognitive function. Patients with cancer pain who were consecutively admitted to two different hospice centers, an acute care consultation service in a teaching hospital or a tertiary palliative care unit in a second teaching hospital were evaluated for study entry. Two independent palliative care specialists completed the rESS where possible within 24 hours of each other. Patients' pain ratings and opioid consumption were recorded daily until the study endpoint (i.e. achievement of stable pain control, discharge or death). Seven hundred and forty-six patients were eligible for study entry and of these, 619 (83%) had a pain syndrome. Inter rater reliability estimates ranged from 0.67 (pain mechanism) to 0.95 (presence of addiction). In the univariate Cox regression analysis, younger patients (<60), as well as patients with neuropathic pain, incidental pain, psychological distress, or co-morbid psychological distress and addiction, required a significantly longer time to achieve stable pain control (P<0.05). In the multivariate Cox regression analysis, only age (<60), neuropathic pain and incidental pain were significantly associated with time to reach stable pain control (PA (p.A324T) variant located in the substrate binding domain is likely a mut- mutation. The remaining missense mutations c.326A>G (p.Q109R), c.983T>C (p.L328P), c.1846C>T (p.R616C), and c.1850T>G (p.L617R) are probably mut0. In the MMAA patients analyzed, frameshift mutations are prevalent. We have explored the genotype phenotype correlation for this clinically heterogeneous disease. PMID- 15781193 TI - Thymidine phosphorylase gene mutations in patients with mitochondrial neurogastrointestinal encephalomyopathy syndrome. AB - The mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) syndrome is characterized by the association of gastrointestinal and neurological symptoms. It is a rare autosomal recessive mitochondrial disorder with multiple mitochondrial DNA deletions and/or depletion. It is caused by thymidine phosphorylase (TP) gene mutations resulting in a complete abolition of TP activity. We tested 31 unrelated patients presenting either with a complete MNGIE syndrome (8 patients), a severe intestinal pseudo-obstruction (10 patients), and multiple deletions and/or depletion of mitochondrial DNA (13 patients). All the tested patients presenting with a complete MNGIE had increased thymidine levels in plasma and urine, and no TP activity. The group with pseudo-obstruction syndrome had normal or partial reduction of TP activity. We found pathogenic mutations on TP gene only in the MNGIE syndrome group: all the MNGIE patients were compound heterozygous or homozygous for mutations in the TP gene. Eight of these mutations are yet unreported, confirming the lack of genotype/phenotype correlation in this syndrome. Enzymatic activity and thymidine level are thus rapid diagnosis tests to detect MNGIE affected patients prior to genetic testing for patients with gastrointestinal symptoms. PMID- 15781194 TI - GALC transduction leads to morphological improvement of the twitcher oligodendrocytes in vivo. AB - Globoid cell leukodystrophy (GLD, Krabbe disease) is a severe demyelinating disease caused by a genetic defect of beta-galactocerebrosidase (GALC). To date treatment to GLD is limited to hematopoietic stem cell transplantation. Experimental approaches by means of gene therapy in twitcher mouse, an authentic murine model of human GLD, showed significant but only marginal improvements of the disease. To clarify whether the introduction of GALC could provide beneficial effects on the oligodendrocytes in GLD, we transduced twitcher oligodendrocytes by stereotactically injecting recombinant retrovirus encoding GALC-myc-tag fusion gene into the forebrain subventricular zone of neonatal twitcher mouse. In vivo effects of exogenous GALC on twitcher oligodendrocytes were studied histologically by combined immunostaining for the myc-epitope and the oligodendroglial specific marker, pi form of glutathione-S-transferase, at around 40 days of age. We show here that GALC transduction led to dramatic morphological improvement of the twitcher oligodendrocytes comparing with those in untreated twitcher controls. This study provided direct in vivo evidence that GALC transduction could prevent or correct aberrant morphology of oligodendrocytes in GLD which may be closely related to the dysfunction and/or degeneration of oligodendrocytes and the demyelination in this disease. PMID- 15781195 TI - Common polymorphisms in the PPARgamma2 and IRS-1 genes and their interaction influence serum adiponectin concentration in young Finnish men. AB - The Gly972Arg substitution of the insulin receptor substrate-1 (IRS-1) gene and the Pro12Pro genotype of the peroxisome proliferator-activated receptor gamma 2 (PPARgamma2) gene have been suggested to be associated with type 2 diabetes mellitus. In this study, the influence of these two polymorphisms on serum adiponectin concentrations was investigated. The Pro12Ala polymorphism of the PPARgamma2 gene and the Gly972Arg polymorphism of the IRS-1 gene were genotyped in 252 young Finnish servicemen. The Ala12Ala genotype of PPARgamma2 was associated with a higher adiponectin level compared to the Pro12Ala genotype (p=0.02) and the Pro12Pro genotype (p=0.02). Total (p=0.02) and low-density lipoprotein (LDL) cholesterol (p=0.03) levels were higher in subjects with the Pro12Pro genotype compared to the Pro12Ala genotype. No difference was observed in serum adiponectin level between the IRS-1 genotype groups. The subjects with X972Arg of this gene had high total and LDL cholesterol levels (p<0.05). The interaction between the PPARgamma2 and IRS-1 genes with respect to their effects on adiponectin levels was statistically significant (p=0.02). Adiponectin was significantly higher (p<0.05) in subjects who simultaneously had the Ala/Ala (PPARgamma2)+Gly/Gly (IRS-1) genotype combination compared to subjects with the Pro/Pro+Gly/Gly and Pro/Ala+Gly/Gly genotype combinations. Total and LDL cholesterol was higher (p<0.05) in subjects with Pro/Pro+X/Arg compared to subjects with the two before mentioned genotype combinations. We conclude that the Ala12Ala genotype of PPARgamma2 is associated with elevated adiponectin level, and that the PPARgamma2 and IRS-1 genes have a possible interaction in their effects on adiponectin concentration. PMID- 15781196 TI - Four new mutations in the BCHE gene of human butyrylcholinesterase in a Brazilian blood donor sample. AB - The genetic variation of human butyrylcholinesterase has been associated with height, body mass index, Alzheimer's disease, and response to xenobiotic agents. The present study reports four new mutations, found in the exon 2 of the BCHE gene, in a sample from 3001 Brazilian blood donors. The three nonsynonymous mutations and one synonymous mutation detected are: 223G-->C, G75R; 270A-->C, E90 D; 297T-->G, I99 M; 486T-->C, A162 A, respectively. All these variants are rare: 0.093+/-0.093% for the missense mutations and 0.137+/-0.137% for the synonymous mutation. A table with the 58 non-usual variants of butyrylcholinesterase is also presented. PMID- 15781197 TI - Clotrimazole decreases human breast cancer cells viability through alterations in cytoskeleton-associated glycolytic enzymes. AB - Cancer cells are characterized by a high rate of glycolysis, which is their primary energy source. Glycolysis is known to be controlled by allosteric regulators, as well as by reversible binding of glycolytic enzymes to cytoskeleton. Clotrimazole is an anti-fungal azole derivative recently recognized as a calmodulin antagonist with promising anti-cancer effect. Here, we show that clotrimazole induced morphological and functional alterations on human breast cancer derived cell line, MCF-7. The drug decreased cell viability in a dose- and time-dependent manner, exhibiting an IC50 of 88.6+/-5.3 microM and a t0.5 of 89.7+/-7.2 min, with 50 microM clotrimazole. Morphological changes were evident as observed by scanning electron microscopy, which revealed the completely loss of protrusion responsible for cell adhesion after a 180 min of treatment with 50 microM clotrimazole. Giemsa stained cells observed by optical microscopy show morphological alterations and a marked nuclear condensation. These changes occurred in parallel to the detachment of the glycolytic enzymes, 6-phosphofructo 1-kinase and aldolase, from cytoskeleton. After a 45 min treatment with 50 microM clotrimazole, the remaining activities in a cytoskeleton enriched fraction was 16.4+/-3.6% and 41.0+/-15.6% of control for 6-phosphofructo-1-kinase and aldolase, respectively. Immunocytochemistry experiments revealed a decrease in the co-localization of 6-phosphofructo-1-kinase and F-actin after clotrimazole treatment, suggesting the site of detachment of the enzymes. Altogether, our results support evidence for apoptotic events that might be started by clotrimazole involving inhibition of glycolytic flux in MCF-7 cells and makes this drug a promising agent in the fight against human breast cancer. PMID- 15781198 TI - Management and outcome of neonatal-onset ornithine transcarbamylase deficiency following liver transplantation at 60 days of life. AB - Neonatal hyperammonemia secondary to X-linked ornithine transcarbamylase (OTC) deficiency carries a high risk of morbidity and mortality. Results of medical therapy are less than satisfactory. Experience with liver transplantation in very young affected infants is limited. We report a male newborn with severe OTC deficiency who underwent successful orthotopic, cadaveric liver transplantation at the age of 60 days. Although technically challenging in the neonatal period, liver transplantation should be considered early as the most promising treatment approach currently available. PMID- 15781199 TI - Mutation analysis of the MCM gene in Korean patients with MMA. AB - Methylmalonic acidemia (MMA) is an autosomal recessive inborn error of metabolism caused by inadequate function of methylmalonyl-CoA mutase. We studied five Korean patients diagnosed with mut MMA, here, we report five new missense mutations (G94E, R369C, S344Y, N189K, and T230I) and a previously reported mutation (R369H) that, this is the first time this mutation has been identified in Korean individuals. Genetic heterogeneity in mut MMA is high. The R369H mutation has been identified in America and Japan. To date, more than 55 different mutations have been identified in mut MMA. A majority of mutations is novel with only three (G717V, E117X, and N219Y) being reported more frequently, the G717V mutation was found in Africa-Americans and Ghanaian. The E117X mutation has been found in Japan. The N219Y mutation has been found in Caucasians and Arab. The R369H mutation is the first mutation identified in three nations (Korea, Japan, and America). PMID- 15781200 TI - Cerebral folate deficiency: life-changing supplementation with folinic acid. AB - Cerebral folate deficiency is characterized by low cerebrospinal fluid (CSF) concentrations of 5-methyltetrahydrofolate and a broad spectrum of clinical signs and symptoms. A patient with progressive spasticity, gait disturbance, speech difficulties, initially diagnosed as a recessive spastic paraplegia recovered on folinic acid (15-30 mg/day) and her 5-methyltetrahydrofolate in CSF normalized. This report demonstrates the importance of CSF investigation in the diagnosis of cerebral folate deficiency and efficiency of folinic acid (5 formyltetrahydrofolate) supplementation. PMID- 15781201 TI - W4R variant in CSRP3 encoding muscle LIM protein in a patient with hypertrophic cardiomyopathy. PMID- 15781202 TI - Analysis of genomic variants in the KCNQ1OT1 transcript in Silver-Russell syndrome patients. PMID- 15781203 TI - Does fluoxetine (Prozak) block mitochondrial permeability transition by blocking VDAC as part of permeability transition pores? PMID- 15781204 TI - Micronucleus formation and induction of apoptosis by different isothiocyanates and a mixture of isothiocyanates in human lymphocyte cultures. AB - Isothiocyanates (ITCs) are the main sulfur-containing metabolites found in cruciferous vegetables. There is evidence that some ITCs may act as chemopreventive agents against different tumor types and induce apoptosis and modulate cell-cycle progression of highly proliferative cancer cells. However, there are also studies reporting genotoxic or co-carcinogenic effects for some ITCs, such as benzyl ITC and phenyl ITC. Since selectivity for transformed cells and absence of genotoxicity for healthy cells are important pre-requisites for new chemopreventive agents, we investigated micronucleus formation and induction of apoptosis by 4-(methylthio)butylisothiocyanate (MTBITC), sulforaphane and a mixture of ITCs in human T-lymphocyte cultures. We demonstrate that MTBITC, sulforaphane and the mixture of ITCs did not induce micronuclei. Moreover, sulforaphane induced a dose-dependent increase in the number of apoptotic cells, which was significant at the highest concentration tested (30 microM) (41% versus 18% in the untreated samples, P<0.05). The mixture of ITCs presented a trend similar to that found for sulforaphane. In fact, the mixture of ITCs was able to induce a dose-dependent increase in the percentage of apoptotic cells, which reached a maximum value at the concentration of 13 microg/ml (46% versus 19% in control samples, P<0.05). Induction of apoptosis was not observed in cultures treated with MTBITC. Our results suggest that different ITCs can have different effects. Moreover, although the mixture of glucosinolates (GLs) used in the present study does not reflect the exact composition of broccoli, our findings demonstrate that the quantitative effects of a single, specific ITC can be significantly different from those of an ITC mixture, where other ITCs of the mixture contribute to the outcome observed. PMID- 15781205 TI - Endoreduplication induced in cultured Chinese hamster cells by different anti topoisomerase II chemicals. Evidence for the essential contribution of the enzyme to chromosome segregation. AB - With the ultimate purpose of testing the hypothesis that, as shown in yeast mutants, any malfunction of DNA topoisomerase II might result in aberrant mitosis due to defective chromosome segregation, we have chosen three chemicals of different nature, recently reported to catalytically inhibit the enzyme. The endpoint selected to assess any negative effect on the ability of topoisomerase II to properly carry out decatenation of fully replicated chromosomes in the G2/M phase of the cell cycle was the presence of metaphases showing diplochromosomes as a result of endoreduplication, i.e. two successive rounds of DNA replication without intervening mitosis. The anti-topoisomerase drugs selected were the anthracycline antibiotic and antineoplastic agent aclarubicin, the respiratory venom sodium azide, and 9-aminoacridine, a chemical compound with planar topology capable of intercalation between DNA bases. Our results show that the three chemicals tested are able to induce endoreduplication to different degrees. These observations seem to lend support to the proposal that topoisomerase II plays a central role in chromosome segregation in mammalian cells. PMID- 15781206 TI - Adsorption of sterigmatocystin by montmorillonite and inhibition of its genotoxicity in the Nile tilapia fish (Oreachromis nilaticus). AB - Sterigmatocystin (Stg) is closely related to the mycotoxin aflatoxin as a precursor in aflatoxin biosynthesis and classified as an IARC Group-2B carcinogen. The aim of this study was to investigate the efficacy of Egyptian montmorillonite (EM), a clay mineral, to adsorb Stg, to test the stability of the resulting complex under different conditions in vitro, and to utilize the Nile tilapia fish as an in vivo model to evaluate the protective effect of EM against Stg-induced toxicity and clastogenicity. In the in vitro study, four concentrations of EM (0.5, 1, 2 and 4 mg/L aqueous solution) and three concentrations of Stg (5, 10 and 50 microg/ml) were tested. The results show that EM had a high capacity of adsorbing Stg at different concentrations tested. The adsorption ranged from 93.1 to 97.8% of the available Stg in aqueous solutions. The complex was stable at different pHs at 37 degrees C in different organic solvents. An in vivo experiment was conducted to evaluate the ability of EM to prevent the toxicity and chromosomal aberrations induced by Stg in the Nile tilapia fish. Fish received an intragastric dose of EM in corn oil (0.5 mg/kg bw) with or without Stg (1.6 microg/kg bw) twice a week for 4 weeks. Body weight was recorded during dosing, and blood and tissue samples were collected at the end of treatment. Stg residues were determined in fish tissue. The results show that Stg was toxic and clastogenic to fish as indicated by the significant decrease of body weight and the increase in frequencies of micronucleated red blood cells (MN RBC) and chromosomal aberrations in the kidney. The intragastric administration of EM combined with Stg to fish resulted in a reduction of the number of MN RBC and the frequency of chromosomal aberrations in the kidney compared with the group treated with Stg alone. It could be concluded that EM itself was safe and successful in the prevention of Stg toxicity and clastogenicity. PMID- 15781207 TI - An in vivo evaluation of induction of abnormal sperm morphology in mice by landfill leachates. AB - Although several reports have demonstrated the acutely toxic and genotoxic effects of landfill leachates in microbial organisms, plants and aquatic animals, the effects of pollutants present in these leachates have not been clarified yet in terrestrial animals. This study mainly aimed to evaluate a potential genetic effect of raw and simulated leachates from Orita-Aperin and Oworonsoki landfills in south-west Nigeria by use of the murine sperm-head abnormality test. These landfills neither have a synthetic membrane liner at the bottom, nor a natural layer of compacted soil with the desired hydraulic conductivity, nor a run-off control system. As a result, the leachates produced are discharged into the environment. Samples designated as Orita-Aperin Raw Leachate (OARL), Orita-Aperin Simulated Leachate (OASL), Oworonsoki Raw Leachate (OWRL) and Oworonsoki Simulated Leachate (OWSL) were analyzed in the sperm-head abnormality test at concentrations (v/v) of 1%, 2.5%, 5%, 10% and 25%. Mice were given 0.5 ml sample per day for five consecutive days by intraperitoneal injection. Each dose group comprised seven mice, and a 5-week exposure period was utilized. The data show that the test mixtures induced a dose-dependent, statistically significant increase (P<0.05) in the number of sperm with abnormal morphology. Physico chemical analysis of the test samples shows that they contained constituents that are capable of inducing mutation in biologic system. The interaction of some of these constituents with the genetic material in the differentiating cells during spermatogenesis may be responsible for these observations. This is relevant in environmental waste management, and for the assessment of the hazardous effects of the chemicals in landfill leachates. PMID- 15781208 TI - In vitro genotoxic effects of the anticancer drug gemcitabine in human lymphocytes. AB - This research was carried out to investigate in vitro genotoxic effects of the anticancer agent gemcitabine on the induction of chromosomal aberrations and sister-chromatid exchange in human lymphocytes. Three doses of gemcitabine (0.001, 0.002 and 0.004 microg/ml) were applied to lymphocyte cultures from 15 donors. There was a significant increase in the induction of chromosome aberrations and in the occurrence of sister-chromatid exchange in these cells. In addition, gemcitabine significantly decreased the mitotic index and replicative index for all doses. Dose-response regression lines were used to compare the individual susceptibilities to gemcitabine with respect to the chromosome aberration and sister-chromatid exchange frequencies. Our results indicate that gemcitabine is able to induce both cytotoxic and genotoxic effects in human lymphocyte cultures in vitro in a dose-dependent manner. PMID- 15781209 TI - Individual responsiveness to induction of micronuclei in human lymphocytes after exposure in vitro to 1800-MHz microwave radiation. AB - The widespread application of microwaves is of great concern in view of possible consequences for human health. Many in vitro studies have been carried out to detect possible effects on DNA and chromatin structure following exposure to microwave radiation. The aim of this study is to assess the capability of microwaves, at different power densities and exposure times, to induce genotoxic effects as evaluated by the in vitro micronucleus (MN) assay on peripheral blood lymphocytes from nine different healthy donors, and to investigate also the possible inter-individual response variability. Whole blood samples were exposed for 60, 120 and 180 min to continuous microwave radiation with a frequency of 1800 MHz and power densities of 5, 10 and 20 mW/cm(2). Reproducibility was tested by repeating the experiment 3 months later. Multivariate analysis showed that lymphocyte proliferation indices were significantly different among donors (p<0.004) and between experiments (p<0.01), whereas the applied power density and the exposure time did not have any effect on them. Both spontaneous and induced MN frequencies varied in a highly significant way among donors (p<0.009) and between experiments (p<0.002), and a statistically significant increase of MN, although rather low, was observed dependent on exposure time (p=0.0004) and applied power density (p=0.0166). A considerable decrease in spontaneous and induced MN frequencies was measured in the second experiment. The results show that microwaves are able to induce MN in short-time exposures to medium power density fields. Our data analysis highlights a wide inter-individual variability in the response, which was confirmed to be a characteristic reproducible trait by means of the second experiment. PMID- 15781210 TI - Green tea consumption, genetic susceptibility, PAH-rich smoky coal, and the risk of lung cancer. AB - Experimental evidence suggests that green tea (Camellia sinesis) may reduce the risk of lung cancer through several hypothesized mechanisms including scavenging oxidative radicals, inhibition of tumor initiation, and modulation of detoxification enzymes. However, epidemiologic results have not been consistent as to the relationship between green tea consumption and lung caner prevention. We employed a population-based case-control study of 122 cases and 122 controls to investigate the effect that green tea consumption may have on the risk of lung cancer and whether polymorphisms in 8-oxoguanine-DNA glycosylase (OGG1), glutathione-S-transferase M1 (GSTM1), and aldo-keto reductase 1C3 (AKR1C3) modify such an association. Daily green tea consumption was associated with a non significant reduction in lung cancer risk. However, the effect of smoky coal exposure was higher for non-drinkers (odds ratio (OR)=4.93; 95% confidence interval (95% CI)=1.27-19.13) than for drinkers (OR=1.88; 95% CI=1.01-3.48). Further, among individuals with the OGG1 Cys(326) allele, daily consumption was associated with a 72% reduction (95% CI=0.09-0.94). Among GSTM1 null homozygotes, those who consumed green tea daily had a non-significant reduction in risk compared with non-consumers. Green tea consumption had no effect among OGG1 Ser(326) homozygotes or GSTM1 carriers. In addition, AKR1C3 genotype did not modulate the effect of green tea consumption. The chemopreventive effects of green tea in this population may be restricted to individuals who are particularly susceptible to oxidative stress and oxidative DNA damage. PMID- 15781212 TI - An association between a NQO1 genetic polymorphism and risk of lung cancer. AB - NAD(P)H:quinone oxidoreductase (NQO1) is a detoxification enzyme that protects against the regeneration of reactive oxygen species chemically induced by oxidative stress, cytotoxicity, mutagenicity, and carcinogenicity. The protection conferred by NQO1 protein reduces certain environmental carcinogens, such as nitroaromatic compounds, heterocyclic amines, and possible cigarette smoke condensate. The gene coding for NQO1 has a genetic polymorphism (C-->T) at nucleotide position 609 (i.e. amino acid codon 187) of the NQO1 cDNA. This polymorphism was shown to reduce NQO1 enzyme activity, thereby diminishing the protection provided by NQO1. Therefore, we hypothesized that individuals with the variant NQO1 genotype are at higher risk for lung cancer. Using a case-control study, we genotyped the NQO1 variants successfully by PCR-RFLP in 826 lung cancer patients and 826 healthy control subjects matched for age, sex, ethnicity, and smoking status. The frequency of the NQO1 T-allele was statistically significantly different among three ethnic groups (p<0.001). In further analysis of Caucasians, the variant NQO1 genotypes (CT and TT) were associated with a marginally increased lung cancer risk (OR=1.19; 95% CI: 0.95-1.50). The elevated lung cancer risk was only evident in younger individuals (age <62 years old) (OR=1.46; 95% CI: 1.04-2.05), women (OR=1.89; 95% CI: 1.33-2.68), and never smokers (OR=1.80; 95% CI: 1.03-3.13). Furthermore, we found a statistically significant trend in the development of lung cancer at an early age in women with increasing copies of the variant allele (p=0.03). These results suggest that the NQO1 variant genotype may modulate lung cancer risk, especially in younger individuals (age<62), women, and never smokers. PMID- 15781213 TI - Chromosomal malsegregation and micronucleus induction in vitro by the DNA topoisomerase II inhibitor fisetin. AB - The plant flavonol fisetin is a common dietary component that has a variety of established biological effects, one of which is the inhibition of the enzyme DNA topoisomerase II (topo II). Compounds that inhibit topo II can exert genotoxic effects such as DNA double strand breaks, which can lead to the induction of kinetochore- or CREST-negative micronuclei. Despite reports that fisetin is an effective topoisomerase II inhibitor, its genotoxic effects have not yet been well characterized. Genotoxicity testing of fisetin was conducted in TK6 and HL60 cell lines and the cells were analyzed for malsegregating chromosomes as well as for the induction of micronuclei. Using the cytokinesis-blocked CREST micronucleus assay to discriminate between micronuclei formed from chromosomal breakage (CREST-negative) and chromosomal loss (CREST-positive), a statistically significant increase in CREST-positive micronuclei was seen for all doses tested in both cell lines. CREST-negative micronuclei, however, were significantly increased at the higher test concentrations in the TK6 cell line. These data indicate that at low concentrations fisetin is primarily exerting its genotoxic effects through chromosomal loss and that the induction of DNA breaks is a secondary effect occurring at higher doses. To confirm these results, the ability of fisetin to inhibit human topoisomerase II-alpha was verified in an isolated enzyme system as was its ability to interfere with chromosome segregation during the anaphase and telophase periods of the cell cycle. Fisetin was confirmed to be an effective topo II inhibitor. In addition, significant increases in the number of mis-segregating chromosomes were observed in fisetin-treated cells from both cell lines. We conclude that fisetin is an aneugen at low concentrations capable of interfering with proper chromosomal segregation and that it is also an effective topo II inhibitor, which exerts clastogenic effects at higher concentrations. PMID- 15781211 TI - DNA damage in lymphocytes of benzene exposed workers correlates with trans,trans muconic acids and breath benzene levels. AB - Benzene causes many kinds of blood disorders in workers employed in many different environments. These diseases include myelodisplastic syndrome and acute and chronic myelocytic leukemia. In the present study, five occupational work places, including six industrial process types, namely, printing, shoe-making, methylene di-aniline (MDA), nitrobenzene, carbomer, and benzene production were selected, and the levels of breath benzene, and trans,trans-muconic acids (t,t MA) and phenol in urine were evaluated, as well as hematological changes and lymphocyte DNA damage. The concentration of benzene in breath was less than 3 ppm in the workplaces, and benzene exposure was found to be higher in work places where benzene is used, than in those where benzene is produced. At low levels of benzene exposure, urinary t,t-MA correlated strongly with benzene in air. Highest Olive tail moments were found in workers producing carbomer. Levels of breathzone benzene were found to be strongly correlated with Olive tail moment values in the lymphocytes of workers, but not with hematological data in the six workplaces types. In conclusion, the highest benzene exposures found occurred in workers at a company, which utilized benzene in the production of carbomer. In terms of low levels of exposure to benzene, urinary t,t-MA and DNA damage exhibited a strong correlation with breath benzene, but not with hematological data. We conclude that breath benzene, t,t-MA and lymphocytic DNA damage are satisfactory biomonitoring markers with respect to benzene exposure in the workplace. PMID- 15781214 TI - Genotoxic effects of eugenol, isoeugenol and safrole in the wing spot test of Drosophila melanogaster. AB - In the present study, the phenolic compounds eugenol, isoeugenol and safrole were investigated for genotoxicity in the wing spot test of Drosophila melanogaster. The Drosophila wing somatic mutation and recombination test (SMART) provides a rapid means to evaluate agents able to induce gene mutations and chromosome aberrations, as well as rearrangements related to mitotic recombination. We applied the SMART in its standard version with normal bioactivation and in its variant with increased cytochrome P450-dependent biotransformation capacity. Eugenol and safrole produced a positive recombinagenic response only in the improved assay, which was related to a high CYP450-dependent activation capacity. This suggests, as previously reported, the involvement of this family of enzymes in the activation of eugenol and safrole rather than in its detoxification. On the contrary, isoeugenol was clearly non-genotoxic at the same millimolar concentrations as used for eugenol in both the crosses. The responsiveness of SMART assays to recombinagenic compounds, as well as the reactive metabolites from eugenol and safrole were considered responsible for the genotoxicity observed. PMID- 15781215 TI - Conversion of tris(8-quinolinolato-N1, O8) aluminum to 8-hydroxyquinoline and activity in bacterial reverse mutation assays. AB - Tris(8-quinolinolato-N1, O8) aluminum (AlQ), an aluminum chelate of 8 hydroxyquinoline (8OHQ) is an important charge transfer molecule in semiconducting imaging devices. This study was conducted to evaluate AlQ and 8OHQ for the ability to induce reverse mutations, either in the presence or absence of mammalian microsomal enzymes, and to determine if AlQ decomposes or is metabolized to 8OHQ under assay conditions. The tester strains used in the mutation assay were Salmonella typhimurium TA98, TA100, TA1535 and TA1537 and Escherichia coli WP2uvrA (pKM101). The assays were conducted in the presence and absence of S9. AlQ doses were 1-1000 microg per plate while 8OHQ doses were 0.947 947 microg per plate to maintain molar equivalency. Stability studies were carried out for 4h at 37 degrees C under conditions designed to mimic mutation assays. Samples were analyzed by HPLC and LC/MS to tentatively identify potential metabolites of AlQ and 8OHQ. The results of the bacterial mutagenicity assay indicate that in the presence of S9, both AlQ and 8OHQ, caused increases in the mean number of revertants per plate with tester strains TA100 and WP2uvrA (pKM101). No increases were observed with any of the remaining tester strain/activation condition combinations. The stability study showed that AlQ degrades readily to 8OHQ under standard mutagenicity test conditions, and the positive test result with AlQ is due to the bioactivation of 8OHQ. In the presence of S9, 8OHQ is metabolized to one detectable product with molecular weight indicative of a one-oxygen insertion. 8OHQ N-oxide and 2,8-quinolinediol were ruled out as possible metabolites; 8OHQ epoxides and other quinolinediols were neither confirmed nor ruled out. Bacterial mutagenicity tests have not been shown to predict in vivo effects of 8OHQ; these assays are similarly expected to be poorly predictive of in vivo genotoxic and carcinogenic potential of AlQ. PMID- 15781216 TI - Nitrogen mustard- and half-mustard-induced damage in Escherichia coli requires different DNA repair pathways. AB - Bifunctional alkylating agents are used in tumor chemotherapy to induce the death of malignant cells through blockage of DNA replication. Nitrogen mustards are commonly used chemotherapeutic agents that can bind mono- or bifunctionally to guanines in DNA. Mustard HN1 is considered a monofunctional analog of bifunctional mustard HN2 (mechlorethamine). Escherichia coli K12 mutant strains deficient in nucleotide excision repair (NER) or base excision repair (BER) were submitted to increasing concentrations of HN2 or HN1, and the results revealed that damage induced by each chemical demands different DNA repair pathways. Damage induced by HN2 demands the activity of NER with a minor requirement of the BER pathway, while HN1 damage repair depends on BER action, without any requirement of NER function. Taken together, our data suggest that HN1 and HN2 seem to induce different types of damage, since their repair depends on distinct pathways in E. coli. PMID- 15781217 TI - Yeast DEL assay detects clastogens. AB - Chromosomal rearrangements, including DNA deletions are involved in carcinogenesis. The deletion (DEL) assay scoring for DNA deletions in the yeast Saccharomyces cerevisiae is able to detect a wide range of carcinogens. Among approximately 60 compounds of known carcinogenic activity, the DEL assay detected 86% correctly whereas the Ames Salmonella assay detected only 30% correctly [R.J. Brennan, R.H. Schiestl, Detecting carcinogens with the yeast DEL assay, Methods Mol. Biol. 262 (2004) 111-124]. Since the DEL assay is highly inducible by DNA double strand breaks, this study examined the utility of the DEL assay for detecting clastogens. Ten model compounds, with varied mechanisms of genotoxicity, were examined for their effect on the frequency of DNA deletions with the DEL assay. The compounds tested were: actinomycin D, camptothecin, methotrexate and 5-fluorodeoxyuridine, which are anticancer agents, noscapine and furosemide are therapeutics, acridine, methyl acrylate and resorcinol are industrial chemicals and diazinon is an insecticide. The in vitro micronucleus assay (IVMN) in CHO cells, a commonly used tool for detection of clastogens, was performed on the same compounds and the results of the two assays were compared. The results of our study show that there is 70% concordance in the presence of metabolic activation (rat liver S9) and 80% concordance in the absence of metabolic activation between the DEL assay and the standard in vitro micronucleus assay. The lack of cytotoxicity observed for four of the ten compounds examined indicates limited diffusion of lipophilic compounds across the yeast cell wall. Thus, the development of a more permeable yeast tester strain is expected to greatly improve concordance of the DEL assay with the IVMN assay. The yeast DEL assay is inexpensive, amenable to automation and requires less expertise to perform than the IVMN. Thus, it has a strong potential as a robust, fast and economical screen for detecting clastogens in vitro. PMID- 15781218 TI - The Arg280His polymorphism in X-ray repair cross-complementing gene 1 impairs DNA repair ability. AB - The contribution of three single nucleotide polymorphisms (SNPs) that substitute amino acids in the X-ray repair cross-complementing gene 1 (XRCC1) protein, Arg194Trp (R194W), Arg280His (R280H), and Arg399Gln (R399Q), to the risk of various types of cancers has been extensively investigated by epidemiological researches. To investigate whether two of these polymorphisms directly influence their repair ability, we established Chinese hamster ovary (CHO) EM9 cell lines transfected with XRCC1(WT), XRCC1(R194W), or XRCC1(R280H) genes and analyzed the DNA repair ability of these cells. The EM9 cells that lack functional XRCC1 proteins exhibit severe sensitivity to methyl methanesulfonate (MMS). Introduction of the human XRCC1(WT) and XRCC1(R194W) gene to EM9 cells restored the MMS sensitivity to the same level as the AA8 cells, a parental cell line. However, introduction of the XRCC1(R280H) gene partially restored the MMS sensitivity, resulting in a 1.7- to 1.9-fold higher sensitivity to MMS compared with XRCC1(WT) and XRCC1(R194W) cells at the LD(50) value. The alkaline comet assay determined diminished base excision repair/single strand break repair (BER/SSBR) efficiency in XRCC1(R280H) cells as observed in EM9 cells. In addition, the amount of intracellular NAD(P)H decreased in XRCC1(R280H) cells after MMS treatment. Indirect immunofluorescence staining of the XRCC1 protein showed an intense increase in the signals and clear foci of XRCC1 in the nuclei of the XRCC1(WT) cells, but a faint increase in the XRCC1(R280H) cells, after MMS exposure. These results suggest that the XRCC1(R280H) variant protein is defective in its efficient localization to a damaged site in the chromosome, thereby reducing the cellular BER/SSBR efficiency. PMID- 15781219 TI - DNA damage and aberrant crypt foci as putative biomarkers to evaluate the chemopreventive effect of annatto (Bixa orellana L.) in rat colon carcinogenesis. AB - Chemoprevention opens new perspectives in the prevention of cancer and other degenerative diseases. Use of target-organ biological models at the histological and genetic levels can markedly facilitate the identification of such potential chemopreventive agents. Colon cancer is one of the highest incidence rates throughout the world and some evidences have indicated carotenoids as possible agents that decrease the risk of colorectal cancer. In the present study, we evaluate the activity of annatto (Bixa orellana L.), a natural food colorant rich in carotenoid, on the formation of aberrant crypt foci (ACF) induced by dimethylhydrazine (DMH) in rat colon. Further, we investigate, the effect of annatto on DMH-induced DNA damage, by the comet assay. Male Wistar rats were given s.c. injections of DMH (40 mg/kg body wt.) twice a week for 2 weeks to induce ACF. They also received experimental diets containing annatto at 20, 200 or 1000 ppm for five 5 weeks before (pre-treatment), or 10 weeks after (post treatment) DMH treatment. In both protocols the rats were sacrificed on week 15th. For the comet assay, the animals were fed with the same experimental diets for 2 weeks. Four hours before the sacrifice, the animals received an s.c. injection of DMH (40 mg/kg body wt.). Under such conditions, dietary administration of 1000 ppm annatto neither induce DNA damage in blood and colon cells nor aberrant crypt foci in rat distal colon. Conversely, annatto was successful in inhibiting the number of crypts/colon (animal), but not in the incidence of DMH-induced ACF, mainly when administered after DMH. However, no antigenotoxic effect was observed in colon cells. These findings suggest possible chemopreventive effects of annatto through their modulation of the cryptal cell proliferation but not at the initiation stage of colon carcinogenesis. PMID- 15781221 TI - Strange phenomenon, i.e., an antimutagenic effect of areca nut chewing. PMID- 15781220 TI - Protection by quercetin and quercetin-rich fruit juice against induction of oxidative DNA damage and formation of BPDE-DNA adducts in human lymphocytes. AB - Flavonoids are claimed to protect against cardiovascular disease, certain forms of cancer and ageing, possibly by preventing initial DNA damage. Therefore, we investigated the protective effects of the flavonoid quercetin against the formation of oxidative DNA damage and bulky DNA adducts in human lymphocytes, both in vitro and ex vivo. First, human lymphocytes were pre-incubated with various concentrations of quercetin, followed by incubation with hydrogen peroxide; protection against oxidative DNA damage was evaluated by use of the single-cell gel electrophoresis (Comet) assay. Second, quercetin-treated human lymphocytes were challenged by treatment with benzo(a)pyrene (B(a)P), and BPDE DNA adduct formation was measured by (32)P-postlabelling. Third, in a pilot study, lymphocytes from female volunteers who consumed a quercetin-rich blueberry/apple juice mixture for four weeks, were treated ex vivo with an effective dose of H(2)O(2) and benzo(a)pyrene, respectively, at three different time points, i.e. before (t=0 weeks), during (t=2 weeks) and after (t=4 weeks) the intervention. Results in vitro: a significant dose-dependent protection by quercetin against both the formation of oxidative DNA damage (p<0.01) and of BPDE DNA adducts (p<0.05) was observed. Results in vivo: four weeks of juice intervention led to a significant increase in the total antioxidant capacity of plasma, as reflected by the increase of the TEAC value from 773 microM trolox equivalent at t=0 to 855 microM at t=4 weeks (p=0.04) and an increase in plasma quercetin content from 5.0 to 10.6 nM (p=0.03). After intervention, the level of oxidative damage upon ex vivo exposure to H(2)O(2) was non-significantly (p=0.07) decreased by 41%, and the BPDE-DNA adduct level induced ex vivo was non significantly decreased by 11%. The combination of our findings in vitro and ex vivo provides evidence that quercetin is able to protect against chemically induced DNA damage in human lymphocytes, which may underlie its suggested anticarcinogenic properties. PMID- 15781223 TI - Collagen overglycosylation: a biochemical feature that may contribute to bone quality. AB - Skeletal ability to resist mechanical stress is determined by bone amount and quality, which relies on macro- and micro-architecture, turnover, bone matrix, and mineralisation; the role of collagen has not been clearly elucidated. Numerous post-translational steps are involved in collagen type I biosynthesis, including residue hydroxylation and glycosylation catalysed by enzymes that work until the protein folds forming the triple helix; therefore, folding rate regulates these processes. Overglycosylated hydroxylysines are poor substrates for epsilon-amino group deamination which initiates cross-link formation. Three clinical conditions associated with fractures may relate collagen overglycosylation with bone quality: (i) Osteogenesis Imperfecta, in which genetic mutations distort triple helix conformation and slow folding rate favouring overglycosylation; (ii) diabetes mellitus, with collagen overglycosylation by AGE accumulation; and, (iii) menopause, according to experimental studies demonstrating ovariectomy-related trabecular bone collagen overglycosylation preventable by 17beta-estradiol or tamoxifen. Specific actions on collagen of drugs used for bone protection should be explored in future studies. PMID- 15781224 TI - Scratch-wounding renders cultivated cells less permissive to prion infection. AB - Using permissive cell lines of epithelial or neuroglial origin, we found that scratch-wounding a small proportion of the recipient cells prior to prion exposure strongly reduced the cell culture's susceptibility to infection. We provide evidence suggesting that wound-triggered inhibition of prion infection was mediated by the release of nucleotides in the extracellular medium of injured cultures. While cell wounding or ATP treatment of unwounded target cells inhibited de novo infection, we found that they had no effect on steady-state infected cultures, indicating that these treatments affect the early stages of infection. These findings support the view that cells have the capacity to modulate their permissiveness to prion infection in response to external stimuli, such as a signalling molecule. PMID- 15781225 TI - Inhibition of the DNA binding by the TCF-1 binding RNA aptamer. AB - The DNA binding architectural protein, TCF, and the transcriptional activator, beta-catenin, form a complex that regulates the expression of diverse target genes during early development and carcinogenesis. As an approach to modulating transcription by this complex, we selected an RNA aptamer that binds to the DNA binding domain of TCF-1. The aptamer interfered with the binding of TCF-1 to its specific DNA recognition sequences in vitro and also inhibited DNA binding of cellular TCF-1. We also developed the truncated version of the aptamer for efficient delivery to the cells. Structural analysis of the truncated aptamer revealed that a stem-loop with an internal loop was responsible for the binding to TCF-1. Similar approach may well be applicable to other proteins, especially DNA binding transcription factors, in order to modulate their DNA binding and transcriptional activity in the cells. PMID- 15781226 TI - Circadian expression of clock genes and clock-controlled genes in the rat retina. AB - The circadian expression patterns of genes encoding for proteins that make up the core of the circadian clock were measured in rat retina using real-time quantitative PCR (qPCR). Transcript levels of several genes previously used for normalization of qPCR assays were determined and the effect of ischemia reperfusion on the expression of clock genes was studied. Statistically significant circadian changes in transcript levels were found for: Per2, Per3, Cry2, Bmal1, Rora, Rorb, and Rorc with changes ranging between 1.6- and 2.6-fold. No changes were found for Per1, Cry1, Clock, Rev-erb alpha, and Rev-erb beta. Significant differences in transcript levels were observed for several candidate reference genes: HPRT, GAPDH, rhodopsin, and Thy1 and, consequently, the use of these genes for normalization purposes in qPCR or Northern blots may lead to erroneous conclusions. Ischemia-reperfusion leads to a persistent decrease of Per1 and Cry2, which may be related to the selective degeneration of amacrine and ganglion cells. We conclude that while all clock genes are expressed in the retina, only a few show a clear circadian pattern. PMID- 15781227 TI - Activation of PI3K-Akt-GSK3beta pathway mediates hepatocyte growth factor inhibition of RANTES expression in renal tubular epithelial cells. AB - Hepatocyte growth factor (HGF) was recently reported to ameliorate renal inflammation in a rat model of chronic renal failure. HGF exerted its action through suppression of RANTES expression in renal tubules. In the present study, we utilized an in vitro model of human kidney proximal tubule epithelial cells (HKC) to elucidate the mechanisms of RANTES suppression by HGF. HGF significantly suppressed basal and TNF-alpha-induced mRNA and protein expression of RANTES in a time and dose dependent fashion. HGF elicited PI3K-Akt activation and inhibited GSK3, a downstream transducer of PI3K-Akt, by inhibitory phosphorylation at Ser 9. When the PI3K-Akt pathway was blocked by wortmannin, HGF inhibition of RANTES was abrogated, demonstrating that the PI3K-Akt pathway is necessary for HGF action. In addition, specific inhibition of GSK3 activity by lithium ion suppressed basal and TNF-alpha-induced RANTES expression, reminiscent of the action of HGF. To further investigate the role of GSK3 in modulating RANTES expression, we examined the effect of forced expression of wild type GSK3beta or an uninhibitable mutant GSK3beta, in which the regulatory Ser-9 residue is changed to alanine (S9A-GSK3beta) in HKC. Overexpression of wild type GSK3beta did not alter the inhibitory action of HGF on RANTES. In contrast, expression of S9A-GSK3beta abolished HGF inhibition of basal and TNF-alpha stimulated RANTES expression. These findings suggest that PI3K-Akt activation and subsequent inhibitory phosphorylation of GSK3beta are required for HGF-induced suppression of RANTES in HKC. PMID- 15781228 TI - Preserved synaptic vesicle recycling in hippocampal neurons in a mouse Alzheimer's disease model. AB - A recently described triple-transgenic mouse model (3xTg, PS1(M146V), APP(Swe), and tau(P301L)) develops a neuropathology similar to the brains of Alzheimer's disease patients including progressive deposits of plaques and tangles [Neuron 39 (2003) 409]. These mice also show age-related deficits in hippocampal synaptic plasticity that occurs before the development of plaques and tangles. Here we report unchanged synaptic vesicle recycling, as measured by FM1-43 release, in the hippocampal neurons of the 3xTg mice. Expression levels of presynaptic protein synaptophysin and of proteins involved in synaptic vesicle recycling including AP180, dynamin I, and synaptotagmin I also remain unaffected. These data suggest that the synaptic deficits observed in the 3xTg neurons may not arise from the preserved synaptic vesicle recycling. PMID- 15781229 TI - Design of recombinant protein-based SARS-CoV entry inhibitors targeting the heptad-repeat regions of the spike protein S2 domain. AB - Entry of SARS-CoV into a target cell is initiated by binding of the S1 domain of spike protein to a receptor, followed by conformational changes of the spike protein S2 domain, resulting in the formation of a six-helix bundle by the heptad repeat (HR1 and HR2) regions. Our previous studies have demonstrated that peptides derived from HR2 region could inhibit SARS-CoV entry. However, synthesis of these peptides is at high cost. In this study, we designed two recombinant proteins, one containing two HR1 and one HR2 peptides (denoted HR121), and the other consisting of two HR2 and one HR1 peptides (designated HR212). These two proteins could be easily purified with the low cost of production, exhibiting high stability and potent inhibitory activity on entry of the HIV/SARS pseudoviruses with IC(50) values of 4.13 and 0.95muM, respectively. These features suggest that HR121 and HR212 can serve as potent inhibitors of SARS-CoV entry. PMID- 15781230 TI - Ganoderma lucidum suppresses angiogenesis through the inhibition of secretion of VEGF and TGF-beta1 from prostate cancer cells. AB - Ganoderma lucidum (G. lucidum) is a popular medicinal mushroom that has been used as a home remedy for the general promotion of health and longevity in East Asia. The dried powder of G. lucidum, which was recommended as a cancer chemotherapy agent in traditional Chinese medicine, is currently popularly used worldwide in the form of dietary supplements. We have previously demonstrated that G. lucidum induces apoptosis, inhibits cell proliferation, and suppresses cell migration of highly invasive human prostate cancer cells PC-3. However, the molecular mechanism(s) responsible for the inhibitory effects of G. lucidum on the prostate cancer cells has not been fully elucidated. In the present study, we examined the effect of G. lucidum on angiogenesis related to prostate cancer. We found that G. lucidum inhibits the early event in angiogenesis, capillary morphogenesis of the human aortic endothelial cells. These effects are caused by the inhibition of constitutively active AP-1 in prostate cancer cells, resulting in the down regulation of secretion of VEGF and TGF-beta1 from PC-3 cells. Thus, G. lucidum modulates the phosphorylation of Erk1/2 and Akt kinases in PC-3 cells, which in turn inhibits the activity of AP-1. In summary, our results suggest that G. lucidum inhibits prostate cancer-dependent angiogenesis by modulating MAPK and Akt signaling and could have potential therapeutic use for the treatment of prostate cancer. PMID- 15781231 TI - Simple and efficient DNA vector-based RNAi systems in mammalian cells. AB - We have constructed four different RNA polymerase III (Pol III)-based expression vectors, containing H1 or U6 promoters from human and mouse, which enable the endogenous production of small RNA transcripts for gene silencing applications. In addition, to facilitate the selection of recombinant clones, we have further improved these vectors by constructing a stuffer of puromycin resistance gene (Puro(r)) between ClaI and HindIII sites, which makes the preparation of vectors easy for rapid and efficient cloning of targeting sequences. A comparative analysis of the silencing efficiency between shRNA, sense-RNA, antisense-RNA, and siRNA showed that both the shRNA and siRNA, but not the sense-RNA and antisense RNA, dramatically inhibit the targeting gene firefly luciferase activity in mammalian cells. However, there were no significant differences in the inhibition of firefly luciferase expression by shRNA and siRNA expressed from these DNA vectors. In summary, these improved DNA vector-based RNAi systems should provide a simple, convenient, and efficient cloning strategy for studying gene functions in mammalian cells. PMID- 15781232 TI - Sulfur K-edge X-ray absorption spectroscopy as an experimental probe for S nitroso proteins. AB - X-ray absorption spectroscopy at the sulfur K-edge (2.4-2.6keV) provides a sensitive and specific technique to identify S-nitroso compounds, which have significance in nitric oxide-based cell signaling. Unique spectral features clearly distinguish the S-nitroso-form of a cysteine residue from the sulfhydryl form or from a methionine thioether. Comparison of the sulfur K-edge spectra of thiolate, thiol, thioether, and S-nitroso thiolate compounds indicates high sensitivity of energy positions and intensities of XAS pre-edge features as determined by the electronic environment of the sulfur absorber. A new experimental setup is being developed for reaching the in vivo concentration range of S-nitroso thiol levels in biological samples. PMID- 15781234 TI - Calcitonin receptor-stimulating peptide-1 regulates ion transport and growth of renal epithelial cell line LLC-PK1. AB - Calcitonin receptor-stimulating peptide-1 (CRSP-1) is a peptide recently identified from porcine brain by monitoring the cAMP production through an endogenous calcitonin (CT) receptor in the renal epithelial cell line LLC-PK(1). Here we investigated the effects of CRSP-1 on the ion transport and growth of LLC PK(1) cells. CRSP-1 inhibited the growth of LLC-PK(1) cells with a higher potency than porcine CT. CRSP-1 enhanced the uptake of (22)Na(+) into LLC-PK(1) cells more strongly than did CT and slightly reduced the (45)Ca(2+) uptake. The enhancement of the (22)Na(+) uptake was abolished by 5-(N-ethyl-N-isopropyl) amiloride, a strong Na(+)/H(+) exchanger (NHE) inhibitor for NHE1, even at a concentration of 1x10(-8)M, although other ion transporter inhibitors did not affect the (22)Na(+) uptake. These results indicate that CRSP-1 enhances the (22)Na(+) uptake by the specific activation of NHE1. Taken together, CRSP-1 is considered to be a new regulator for the urinary ion excretion and renal epithelial cell growth. PMID- 15781233 TI - Absolute mRNA levels and transcriptional regulation of the mouse testis-specific thioredoxins. AB - Thioredoxins function as general protein disulphide reductases. Mammalian male germ cells are equipped with a set of three testis-specific thioredoxins (named Sptrx-1, -2, and -3, respectively) that are expressed either in different structures within the sperm cell or at different stages of sperm development. Previous studies based on qualitative northern-blot and in situ hybridization analyses restricted the presence of Sptrx mRNAs to adult testis, but nothing is known about their transcriptional regulation or relative expression levels in this tissue. In this report, we investigate the transcriptional profiles of the mouse Sptrx genes in terms of the germ cell-specific regulation by promoter analysis in GC-2spd(ts) cells. Besides, we perform a comprehensive quantification of the Sptrx mRNA molecules by real-time PCR in whole-animal experiments. By these means, we show that transcription is differentially regulated for each Sptrx gene and identify the 5'-flanking regions anticipated to contain the cis regulatory elements responsible, at least in part, for the transcriptional silencing and/or activation of the Sptrx genes. In addition, we show remarkable age-associated variations between the Sptrx mRNA expression patterns. PMID- 15781235 TI - A proteomics approach to identify changes in protein profiles in pre-cancerous colon. AB - The development of colon cancer is characterised by alterations in multiple genetic and epigenetic pathways in colon tissue leading ultimately to deregulation of colon epithelial cells. Early detection is an important factor in decreasing colon cancer deaths. Proteomic techniques were used to identify potential early markers in colon tissue exhibiting pre-cancerous activity that may characterise pathological changes in a chemically induced colon cancer rat model. Protein profiles were assessed in soluble and insoluble fractions prepared from distal colon of rats treated with the colonotropic carcinogen, dimethylhydrazine. Alterations in protein profiles were associated with the presence of aberrant crypt foci, hyperplasia and dysplasia, microanatomical changes, and metabolic changes in rat colon. These changes may have a potential role in the identification of pre-pathological features preceding colon tumorigenesis. PMID- 15781236 TI - Caveolin-2 regulation of the cell cycle in response to insulin in Hirc-B fibroblast cells. AB - The regulatory function of caveolin-2 in cell cycle regulation by insulin was investigated in human insulin receptor-overexpressed rat 1 fibroblast (Hirc-B) cells. Insulin increased induction of the caveolin-2 gene in a time-dependent manner. Direct interaction between ERK and caveolin-2 was confirmed by immunoprecipitation and phosphorylated ERK increased the specific interaction in response to insulin. That insulin induced their nuclear co-localization over time was demonstrated by immunofluorescence microscopy. Insulin increased the S phase in the cell cycle by 6-fold. When recombinant caveolin-1 was transiently expressed, a decrease in the S phase was detected by flow-cytometry. The results indicate that the up-regulation of caveolin-2 in response to insulin activates the downstream signal cascades in the cell cycle, chiefly the increased phosphorylation of ERK, the nuclear translocation of phosphorylated ERK, and the subsequent activation of G0/G1 to S phase transition of the cell cycle. The results also suggest that DNA synthesis and the activation of the cell cycle by insulin are achieved concomitantly with an increase in the interaction between caveolin-2 and phosphorylated ERK, and the nuclear translocation of that complex. Taken together, we conclude that caveolin-2 positively regulates the insulin induced cell cycle through activation of and direct interaction with ERK in Hirc B cells. PMID- 15781237 TI - N-terminal helix reorients in recombinant C-fragment of Clostridium botulinum type B. AB - Botulinum neurotoxins comprise seven distinct serotypes (A-G) produced by Clostridium botulinum. The crystal structure of the binding domain of the botulinum neurotoxin type B (BBHc) has been determined to 2A resolution. The overall structure of BBHc is well ordered and similar to that of the binding domain of the holotoxin. However, significant structural changes occur at what would be the interface of translocation and binding domains of the holotoxin. The loop 911-924 shows a maximum displacement of 14.8A at the farthest point. The N terminal helix reorients and moves by 19.5A from its original position. BBHc is compared with the binding domain of the holotoxin of botulinum type A and B, and the tetanus C-fragment to characterize the heavy chain-carbohydrate interactions. The probable reasons for different binding affinity of botulinum and tetanus toxins are discussed. PMID- 15781238 TI - Loss of lysophospholipase 3 increases atherosclerosis in apolipoprotein E deficient mice. AB - Human LCAT-like lysophospholipase (LLPL), or lysophospholipase 3, was first identified in vitro, in foam cells derived from THP-1 cells. We demonstrated that LLPL was present in foam cells in the severe atherosclerotic lesions that develop in apolipoprotein E-null (apoE(-/-)) mice. This indicated that LLPL might affect lipid metabolisms in foam cells and, therefore, atherogenesis. Accordingly, we created LLPL-knockout mice by gene targeting and crossed them with apoE(-/-) mice. We showed that the absence of LLPL increased lesion formation markedly in apoE(-/-) mice but had little effect on the plasma-lipid profile. In addition, LLPL-deficient peritoneal macrophages were more sensitive to apoptosis induced by exposure to oxidized low-density lipoprotein. LLPL might provide a link between apoptosis in macrophages and atherogenesis. Our data demonstrate that LLPL activity is anti-atherogenic and indicate that the regulation of this enzyme might be a novel drug target for the treatment of atherosclerosis. PMID- 15781239 TI - Preformed beta-amyloid fibrils are destabilized by coenzyme Q10 in vitro. AB - Inhibition of the formation of beta-amyloid fibrils (fAbeta), as well as the destabilization of preformed fAbeta in the CNS, would be attractive therapeutic targets for the treatment of Alzheimer's disease (AD). We reported previously that nordihydroguaiaretic acid (NDGA) and wine-related polyphenol, myricetin (Myr), inhibit fAbeta formation from Abeta and destabilize preformed fAbeta in vitro. Using fluorescence spectroscopic analysis with thioflavin T and electron microscopic studies, we examined the effects of coenzyme Q(10) (CoQ(10)) on the formation, extension, and destabilization of fAbeta at pH 7.5 at 37 degrees C in vitro. We next compared the anti-amyloidogenic activities of CoQ(10) with NDGA and Myr. CoQ(10) dose-dependently inhibited fAbeta formation from amyloid beta peptide (Abeta), as well as their extension. Moreover, it destabilized preformed fAbetas. The anti-amyloidogenic effects of CoQ(10) were slightly weaker than those of NDGA and Myr. CoQ(10) could be a key molecule for the development of therapeutics for AD. PMID- 15781240 TI - Isolation of wound healing/regeneration genes using restrictive fragment differential display-PCR in MRL/MPJ and C57BL/6 mice. AB - Wound healing in mammals can take several weeks to months and the process is always accompanied by scar formation. Wound healing mechanisms that mimic regeneration are not found in most mature mammalian tissues. However, the MRL/MPJ (MRL) mouse has the unique capacity to regenerate ear hole wound completely in less than a month. To identify genes involved in wound healing without a scar, we chose to use restriction fragment differential display-PCR to isolate genes differentially expressed in the MRL (good healer) mouse and the C57BL/6 (poor healer) mouse at different stages of wound healing. We identified 36 genes that were differentially expressed in the regenerating tissue of good and poor healer strains of which several genes are also genetically linked to wound healing and thus are potential candidate genes for scarless wound healing. PMID- 15781241 TI - Mechanical force activates eIF-2alpha phospho-kinases in fibroblast. AB - Mechanical forces can induce differentiation of fibroblasts into myofibroblasts, a process which requires activation of the MAP kinase p38. Currently, the identification of other phospho-kinases involved in myofibroblast differentiation has not been explored. We applied static tensile forces to rat cardiac fibroblasts via collagen-coated magnetite beads and examined activation of protein phospho-kinases by the Kinexus phospho-antibody screening system. Of 75 candidate protein kinases screened, 39 were detected and, of these, 31 phospho kinases were analyzed. Following force application, 12 out of 31 phospho-kinases exhibited increases of phosphorylation including PKR (>4-fold), MKK3 (3-fold), MKK6 ( approximately 2-fold), and p38 ( approximately 2-fold). In several types of mechanically sensitive, contractile fibroblasts including rat cardiac, human gingival, and Rat-2 fibroblasts, tensile forces increased eIF-2alpha phosphorylation, a downstream effector of PKR. We conclude that phospho-antibody screening is an efficient method for discovery of novel mechanical force-induced phospho-kinases and force can activate eIF-2alpha phospho-kinases in fibroblasts. PMID- 15781242 TI - Oxygenation alters ganglioside expression in rat liver following partial hepatectomy. AB - Gangliosides from livers of weanling rats were analyzed after 15% partial hepatectomy (PH) and different pre- and post-operative hyberbaric oxygenation (pre- and postHBO). Neu5Ac was the predominant ganglioside-derived sialic acid (>85%) compared to Neu5Gc. Almost identical low total sialic acid content (Neu5Ac+Neu5Gc) of the control and operated nonHBO animals opposed a 6.4- to 7.6 fold increase in pre- and postHBO animals (69.26 and 81.64pmol/mg wet weight, respectively). NanoESI-QTOF mass spectrometry combined with HPTLC immunostaining revealed GM3(Neu5Ac) and GM3(Neu5Gc) as major gangliosides, correlating with the respective sialic acid concentrations. Minor neolacto-series gangliosides were enhanced in preHBO and postHBO, but GM1-core gangliosides only in preHBO rats. GM2 and GalNAc-GM1b were clearly detectable in oxygenated rats compared to traces in the control and nonHBO animals. These results point at a functional role of gangliosides in liver growth regulation and reconstitution after PH combined with pre- and post-operative HBO treatment. PMID- 15781243 TI - Epithelial differentiation of human adipose tissue-derived adult stem cells. AB - Adult human stem cells are employed in novel treatments and bio-artificial devices. Recent studies have identified an abundant source of stem cells termed adipose-derived adult stem (ADAS)-cells in the subcutaneous adipose tissue. Under appropriate culture conditions ADAS-cells differentiate to various cell types, including chondrocytes, adipocytes, and smooth muscle cells. Aiming at epithelial differentiation this study investigated the effect of all-trans retinoic acid (ATRA) on human ADAS-cells. ATRA-induced cytokeratin 18 expression in ADAS-cells and nearly abolished vimentin expression as shown by Western blot. In immunofluorescence, the formation of keratin fibers in ATRA-treated ADAS-cells could be observed. The percentage of ADAS-cells being able to undergo epithelial differentiation as quantified by FACS-analysis was above 80%. Inhibition of cell growth by ATRA was shown using DAPI- and MTT-assays. ATRA can differentiate ADAS cells toward the epithelial lineage. This finding, along with a previously described neural differentiation, shows that ADAS-cells have epithelial potential. PMID- 15781244 TI - Aconitase and ATP synthase are targets of malondialdehyde modification and undergo an age-related decrease in activity in mouse heart mitochondria. AB - The main purpose of this study was to identify mitochondrial proteins that exhibit post-translational oxidative modifications during the aging process and to determine the resulting functional alterations. Proteins forming adducts with malondialdehyde (MDA), a product of lipid peroxidation, were identified by immunodetection in mitochondria isolated from heart and hind leg skeletal muscle of 6-, 16-, and 24-month-old mice. Aconitase, very long chain acyl coenzyme A dehydrogenase, ATP synthase, and alpha-ketoglutarate dehydrogenase were detected as putative targets of oxidative modification by MDA. Aconitase and ATP synthase from heart exhibited significant decreases in activity with age. Very long chain acyl coenzyme A dehydrogenase and alpha-ketoglutarate dehydrogenase activities were unaffected during aging in both heart and skeletal muscle. This suggests that the presence of a post-translational oxidative modification in a protein does not a priori reflect an alteration in activity. The biological consequences of an age-related decrease in aconitase and ATP synthase activities may contribute to the decline in mitochondrial bioenergetics evident during aging. PMID- 15781245 TI - Cloning and characterization of a lectin from the octocoral Sinularia lochmodes. AB - In the present study, the entire amino acid sequence and cDNA structure encoding the d-galactose-binding lectin, SLL-2, isolated from the octocoral Sinularia lochmodes, were determined. SLL-2 regulates the morphology of symbiotic dinoflagellates Symbiodinium spp. through unknown mechanisms. Here, three cDNAs that encode SLL-2 were cloned and characterized. All the SLL-2 cDNAs encoded 142 amino acids with high similarity to each other. The mature subunit of SLL-2 was found to be composed of 94 amino acids and to contain one putative glycosylation site common to all three SLL-2. N-Glycopeptidase F treatment of SLL-2 resulted in a protein band shift from 16.5 to 9.5kDa in SDS-PAGE, confirming that SLL-2s are glycoproteins. Two-dimensional polyacrylamide gel electrophoresis analysis of the deglycosylated SLL-2 indicated a presence of three polypeptides as encoded in SLL 2 cDNAs. The deduced sequences of SLL-2 cDNAs had a similarity to the C-terminal region of discoidin I, the slime mold Dictyostelium discoideum lectin. PMID- 15781246 TI - Raft-like membrane domains contain enzymatic activities involved in the synthesis of mammalian glycosylphosphatidylinositol anchor intermediates. AB - The synthesis of the glycosylphosphatidylinositol (GPI) anchor occurs in different compartments within the ER. We have previously shown that GPI anchor intermediates including GlcNAc-PI and GlcN-(acyl)PI are present in Triton insoluble membranes (TIMs), believed to be derived from lipid rafts. The present study was initiated to determine if GPI anchor intermediates move to raft-like domains after their synthesis or if these domains represent another ER compartment for GPI anchor synthesis. We determined that in transfected cells Pig Ap and Pig-Lp, two proteins involved in the synthesis of GlcNAc-PI and GlcN-PI, respectively, are present in TIMs. In addition, we detected GlcNAc-PI synthase, GlcNAc-PI deacetylase, and GlcN-PI acyltransferase activities in TIMs isolated from untransfected cells. These results lend support to the possibility of additional GPI biosynthetic compartments in the ER and to the notion that GPI anchor intermediates produced in and outside raft-like domains may have a different fate. PMID- 15781247 TI - Localization of AQP5/AQP8 chimeras in MDCK-II cells: exchange of the N- and C termini. AB - AQP5 and AQP8 possess targeting/retention motifs which mediate their localization to the apical and basolateral membranes, respectively, of polarized MDCK-II cells. As targeting/retention motifs have been localized to the N- or C-termini of other AQPs, we sought the location of such motifs in AQPs 5 and 8 by exchanging their corresponding N- or C-termini and examining the expression, localization, and function of the resultant chimeras. We did not detect the expression of constructs in which the C-terminus of AQP5 was replaced by the C terminus of AQP8. Substitution of the N-terminus of AQP8 for the N-terminus of AQP5 generated a construct which was trapped intracellularly and did not significantly facilitate transepithelial fluid movement. In contrast, modifications of the N- and C-termini of AQP8 were better tolerated. Substitution of either AQP8 terminus by the corresponding AQP5 terminus generated constructs which localized to basolateral membranes and facilitated transepithelial fluid movement. Our results suggest that, unlike the other AQP targeting/retention signals reported thus far, an AQP8 basolateral targeting/retention motif might reside between the two cytosolic termini. PMID- 15781248 TI - Expression and regulation of adrenomedullin in renal glomerular podocytes. AB - Adrenomedullin (AM) is postulated to exert organ-protective effects. It is expressed in the renal glomeruli, but its roles in the glomerular podocytes have been poorly elucidated. In the present study, we investigated the expression and regulation of AM in recently established conditionally immortalized mouse podocyte cell line in vitro and podocyte injury model in vivo. The cultured differentiated podocytes expressed AM mRNA and secreted measurable amount of AM. AM secretion from the podocytes was increased by H(2)O(2), hypoxia, puromycin aminonucleoside (PAN), albumin overload, and TNF-alpha. Real-time RT-PCR analysis revealed that AM mRNA expression in the podocytes was enhanced by PAN and TNF alpha, both of which were suppressed by mitochondrial antioxidants. Furthermore, AM expression was upregulated in the glomerular podocytes of PAN nephrosis rats. These results indicated that AM expression in the podocytes was upregulated by stimuli or condition relevant to podocyte injury, suggesting its potential role in podocyte pathophysiology. PMID- 15781250 TI - Cigarette smoke condensate induces MMP-12 gene expression in airway-like epithelia. AB - Cigarette smoke (CS)-induced emphysema is attributable to matrix metalloproteinase-12 (MMP-12) in mice, however, a relationship between CS and MMP 12 is absent in human emphysema. Here, we show that cigarette smoke condensate (CSC) induces MMP-12 gene expression in airway-like epithelia through a hydrogen peroxide (H(2)O(2))-dependent pathway involving NADPH oxidase, AP-1, and TNF alpha. Cigarette smoke condensate-induced H(2)O(2) production and MMP-12 gene expression were inhibited by apocynin, a specific inhibitor of NADPH oxidases, while 3-aminobenzamide, an inhibitor of AP-1, attenuated CSC-induced MMP-12 gene expression. Messenger RNAs encoding phagocytic NADPH oxidase components and a homologue of p67phox, p51 (NOXA1), were detected, while mRNA of dual oxidase (Duox)1 was unchanged by CSC. Enbrel, an inhibitor of TNF-alpha function, reduced CSC-induced H(2)O(2) production and MMP-12 expression. These findings provide novel evidence of a direct relationship between CS exposure and MMP-12 in human airway epithelia and suggest several targets for modulation of this potentially pathogenic pathway. PMID- 15781249 TI - New antibodies recognizing p73: comparison with commercial antibodies. AB - p73, unlike p53, is expressed as a number of isomeric forms. Alternative splicing at the 3' end of p73 transcript, together with the usage of a second promoter downstream of exon 3, can generate up to 24 p73 isoforms. Variants lacking the TA domain (DeltaN isoforms) are induced by TAp73 and by p53, and inhibit their transcriptional activity. However, understanding the complex biology of p73 has been handicapped by the lack of high affinity specific antibodies for the different isoforms. Here, we report the characterization, by Western blotting and immunoprecipitation, of three new polyclonal antisera recognizing all p73 isoforms, only DeltaN isoforms or only p73alpha, and which have advantages of affinity and specificity over previously available antibodies. PMID- 15781251 TI - Histone deacetylase inhibitor Trichostatin A reduces anti-DNA autoantibody production and represses IgH gene transcription. AB - Systemic lupus erythematosus is characterized by the presence of autoantibodies and hypergammaglobulinemia. To investigate the role of histone deacetylases (HDACs) in the production of autoantibody and immunoglobulin, we examined the effect of Trichostatin A (TSA), a specific inhibitor of HDACs, on anti-DNA autoantibody production and IgH gene transcription. Our results showed that inhibition of HDAC activity by TSA markedly reduced anti-DNA autoantibody production by T347 cells either by inducing apoptosis or in an apoptosis independent manner, suggesting that TSA might be useful for treating certain autoimmune diseases. Moreover, we found that TSA strongly inhibited germline and post-switch immunoglobulin transcripts in T347 cells and in primary splenic B cells of MRL-lpr mice. Reporter gene analysis demonstrated that both Emu and 3' IgH enhancer activities were repressed significantly by TSA-mediated HDAC inhibition. Furthermore, we observed that HDAC1 was recruited to the 3'-IgH enhancer hs1,2 as determined by chromatin immunoprecipitation assays. Over expression of HDAC1 increased the activity of IgH enhancers, especially 3'-IgH enhancers. These findings implicate HDAC in the IgH gene transcription via activation of 3'-IgH enhancers. PMID- 15781252 TI - Characterization of human NIPK (TRB3, SKIP3) gene activation in stressful conditions. AB - The neuronal cell death-inducible putative kinase (NIPK) gene is upregulated in several cell types under stressful conditions. In order to understand the molecular control of the human (h) NIPK gene (also known as TRB3 and SKIP3), we mapped the transcriptional start sites of the gene in HepG2 cells treated with thapsigargin, the inhibitor of endoplasmic reticular Ca(2+)-ATPase, and determined the promoter region of the gene which is essential for endoplasmic reticulum and arsenite stress responses. The analysis of cDNA clones revealed the presence of several hNIPK mRNA isoforms, differing in their 5' regions upstream of the hNIPK translation initiation codon as a result of alternative transcription initiation and alternative splicing. The induction of hNIPK gene in response to thapsigargin and arsenite treatments is mediated by a promoter segment consisting of tandemly arranged 33-bp repeats that contain a regulatory element similar to C/EBP-ATF composite site of the Chop gene promoter. ATF4, whose level is upregulated in the cells exposed to thapsigargin or arsenite, is able to bind to the 33-bp repeat and activate the hNIPK promoter. The coexpression of hNIPK inhibits activation of hNIPK promoter in response to the stress-inducing agents and to overexpressed ATF4, and thus NIPK may function as a negative feedback regulator of ATF4. PMID- 15781253 TI - Identification of a novel estrogen-sulfating cytosolic SULT from zebrafish: molecular cloning, expression, characterization, and ontogeny study. AB - By searching the expressed sequence tag database, a zebrafish cDNA encoding a putative cytosolic sulfotransferase (SULT) was identified. Sequence analysis indicated that this zebrafish SULT belongs to the SULT1 cytosolic SULT gene family. The recombinant form of this novel zebrafish SULT, expressed using the pGEX-2TK expression system and purified from transformed BL21 (DE3) Escherichia coli cells, displayed sulfating activities specifically for estrone and 17beta estradiol among various endogenous compounds tested as substrates. The enzyme also exhibited sulfating activities toward some xenobiotic phenolic compounds. This new zebrafish SULT showed dual pH optima, at 6.5 and 10-10.5, with estrone or n-propyl gallate as substrate. Kinetic constants of the sulfation of estrone, 17beta-estradiol, and n-propyl gallate were determined. Developmental stage dependent expression experiments revealed a significant level of expression of this novel zebrafish estrogen-sulfating SULT at the beginning of the hatching period during embryogenesis, which continued throughout the larval stage onto maturity. PMID- 15781254 TI - Downregulation of IL-12 and a novel negative feedback system mediated by CD25+CD4+ T cells. AB - CD25(+)CD4(+) regulatory T cells suppress immune responses and are believed to play roles in preventing autoimmune diseases. However, the mechanism(s) underlying the suppression and the regulation of their homeostasis remain to be elucidated. Here we show that these regulatory T cells downregulated CD25( )CD4(+) T-cell-mediated production of IL-12 from antigen-presenting cells, which can act as a growth factor for CD25(-)CD4(+) T cells. We further found that CD25(+)CD4(+) T cells, despite their well-documented "anergic" nature, proliferate significantly in vitro only when CD25(-)CD4(+) T cells are present. Notably, this proliferation was strongly dependent on IL-2 and relatively independent of IL-12. Thus, CD25(+)CD4(+) T cells suppress CD25(-)CD4(+) T-cell responses, at least in part, by inhibiting IL-12 production while they themselves can undergo proliferation with the mediation of CD25(-)CD4(+) T cells in vitro. These results offer a novel negative feedback system involving a tripartite interaction among CD25(+)CD4(+) and CD25(-)CD4(+) T cells, and APCs that may contribute to the termination of immune responses. PMID- 15781255 TI - Identification of the human ApoAV gene as a novel RORalpha target gene. AB - Retinoic acid receptor-related orphan receptor-alpha (RORalpha) (NR1F1) is an orphan nuclear receptor with a potential role in metabolism. Previous studies have shown that RORalpha regulates transcription of the murine Apolipoprotein AI gene and human Apolipoprotein CIII genes. In the present study, we present evidence that RORalpha also induces transcription of the human Apolipoprotein AV gene, a recently identified apolipoprotein associated with triglyceride levels. Adenovirus-mediated overexpression of RORalpha increased the endogenous expression of ApoAV in HepG2 cells and RORalpha also enhanced the activity of an ApoAV promoter construct in transiently transfected HepG2 cells. Deletion and mutation studies identified three AGGTCA motifs in the ApoAV promoter that mediate RORalpha transactivation, one of which overlaps with a previously identified binding site for PPARalpha. Together, these results suggest a novel mechanism whereby RORalpha modulates lipid metabolism and implies RORalpha as a potential target for the treatment of dyslipidemia and atherosclerosis. PMID- 15781256 TI - Two glucosylceramide synthase inhibitors attenuate doxorubicin-induced p21Cip1/Waf1 upregulation in HepG2 cells, irrespective of their differential chemosensitizing properties. AB - We have previously reported that HepG2 human hepatocarcinoma cells are sensitized to doxorubicin-induced apoptosis by the glucosylceramide synthase inhibitor d,l threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) but not by the more specific inhibitor d,l-threo-1-phenyl-2-hexadecanoylamino-3-pyrrolidino-1 propanol (PPPP). Herein we investigated whether the chemosensitizing action of PDMP impinged on any unspecific effect of this compound on doxorubicin-induced expression of p53 and/or p21(Cip1/Waf1), namely two proteins reported to modulate the apoptotic response to DNA-damaging agents, in a positive or negative fashion, respectively. We show that, in HepG2 cells, PDMP did not substantially affect doxorubicin-induced p53 upregulation, whereas drug-evoked upregulation of p21(Cip1/Waf1) was markedly attenuated. Although this outcome could be expected to account for the chemosensitizing effect of PDMP, impaired upregulation of p21(Cip1/Waf1), in the setting of unaltered p53 expression, was also observed in the case of PPPP. These results, while raising the possibility of a link between attenuation of drug-evoked p21(Cip1/Waf1) expression and redirection of (glyco)sphingolipid metabolism, show that, differently from other tumor systems, attenuation of doxorubicin-induced p21(Cip1/Waf1) expression is at least not sufficient to sensitize HepG2 cells to the apoptotic action of the drug. PMID- 15781257 TI - Site-specific DNA cleavage by artificial zinc finger-type nuclease with cerium binding peptide. AB - The addition of a new function to native proteins is one of the most attractive protein-based designs. In this study, we have converted a C(2)H(2)-type zinc finger as a DNA-binding motif into a novel zinc finger-type nuclease by connecting two distinct zinc finger proteins (Sp1 and GLI) with a functional linker possessing DNA cleavage activity. As a DNA cleavage domain, we chose an analogue of the metal-binding loop (12 amino acid residues), peptide P1, which has been reported to exhibit a strong binding affinity for a lanthanide ion and DNA cleavage ability in the presence of Ce(IV). Our newly designed nucleases, Sp1(P1)GLI and Sp1(P1G)GLI, can strongly bind to a lanthanide ion and show a unique DNA cleavage pattern, in which certain positions between the two DNA binding sites are specifically cleaved. The present result provides useful information for expanding the design strategy for artificial nucleases. PMID- 15781258 TI - Com-1/P8 in oestrogen regulated growth of breast cancer cells, the ER-beta connection. AB - BACKGROUND: Com-1 is a molecule that has recently discovered to have putative action on the metastatic nature of cancer cells. The current study investigated the impact of Com-1 on oestrogen regulated cell growth of breast cancer cells and explored the potential link between Com-1 and ER-beta. METHOD: Full length Com-1 cDNA was isolated from normal mammary tissues. Ribozyme transgenes that specifically targeted human Com-1 were constructed using the pEF6/V5-His vector. Expression of Com-1 was assessed at both mRNA and protein levels. Interaction of Com-1 with other candidate molecules was studied using immunoprecipitation and Western blotting. RESULTS: Elimination of Com-1 by way of ribozyme transgenes results in breast cancer cells with increased rate of growth and increased invasive potential. In contrast, over-expression of Com-1 in the cancer cells had an opposite effect. In ER-alpha-negative/ER-beta positive MDA MB-231 cells, elimination of Com-1 caused more vigorous growth in response to 17-beta estradiol. However, the effect of Com-1 modification on MCF-7, which is positive for both ER-alpha and ER-beta, was less clear. Protein interaction analysis has indicated that the Com-1 and ER-beta were mutually co-precipitated with each other in breast cancer cells. Immunocytochemical staining revealed that Com-1 was primarily present in the nucleus, with some degree of cytoplasmic staining, and that the distribution of Com-1 was identical to that of ER-beta. 17-Beta estradiol stimulation resulted in reduction of nucleic staining of Com-1. This reduction of nucleic Com-1 can be reverted when ubiquitin inhibitor, ubiquitin aldehyde or the lactacystin proteosome inhibitor was present, suggesting a pivotal role of the ubiquitin-proteosome pathway in the Com-1/ER-beta complex. CONCLUSION: Com-1 plays a tumour suppressor role in breast cancer cells and is involved in oestrogen-regulated cell growth. This action is potentially exerted by interacting with ER-beta, in human breast cancer cells. The fate of Com-1 can be dually regulated by oestrogen and ubiquitin pathway. PMID- 15781259 TI - Expression of B7-H1 and B7-DC on the airway epithelium is enhanced by double stranded RNA. AB - Viral infection in the airway provokes various immune responses, including Th1 and Th2 responses, which are partly initiated by double-stranded RNA (dsRNA), a viral product for its replication. B7-H1 (PD-L1) and B7-DC (PD-L2) are B7-family molecules that bind to programmed death-1 (PD-1) on lymphocytes and are implicated in peripheral tolerance. We investigated the effect of dsRNA on the expression of B7-H1 and B7-DC on airway epithelial cell lines. B7-H1 and B7-DC were constitutively expressed on the cells, and their expression was profoundly upregulated by stimulation with an analog of viral dsRNA, polyinosinic polycytidylic acid. B7-H1 and B7-DC were also upregulated by stimulation with IFN gamma, IL-13, and the supernatant from T cell clones. A relatively high concentration of dexamethasone (1 microM) was required to suppress the upregulation of B7-H1 or B7-DC. These results suggest that epithelial B7-H1 and B7-DC play a role in virus-associated immune responses in the airways. PMID- 15781261 TI - HtrA2 cleaves Apollon and induces cell death by IAP-binding motif in Apollon deficient cells. AB - Apollon/BRUCE is a giant IAP protein that has BIR and UBC domains in its amino- and carboxy-terminals, respectively. Apollon binds and ubiquitylates SMAC/DIABLO and caspase9, and regulates apoptosis by facilitating proteasomal degradation of these proteins. Apollon overexpression inhibits apoptosis, while its downregulation sensitizes cells to apoptosis, suggesting that Apollon level is important for apoptosis regulation. Here we show that HtrA2/Omi catalytically cleaves Apollon with its serine protease activity. Conversely, Apollon ubiquitylates and facilitates proteasomal degradation of HtrA2 that binds to Apollon through IAP-binding motif. Thus, Apollon and HtrA2 mutually downregulate each other. Expression of catalytically active, but not inactive, HtrA2 induced apoptosis in Apollon-expressing cells. In Apollon-deficient cells, however, expression of catalytically inactive HtrA2 mutant with IAP-binding motif also induced apoptosis. These results indicate that HtrA2 induces apoptosis in two different mechanisms, one with serine protease domain and the other with IAP binding motif, in Apollon-deficient cells. PMID- 15781260 TI - Yeast adaptation to 2,4-dichlorophenoxyacetic acid involves increased membrane fatty acid saturation degree and decreased OLE1 transcription. AB - Yeast cells adapted to the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) exhibit a plasma membrane less susceptible to 2,4-D-induced disruption and are more tolerant than unadapted cells to lethal concentrations of the herbicide. These cells, adapted to grow in the presence of increasing concentrations of 2,4 D, were found to exhibit a dose-dependent increase of the saturation degree of membrane fatty acids, associated to the higher percentage of stearic (C(18:0)) and palmitic (C(16:0)) acids, and to the decreased percentage of palmitoleic (Delta9-cisC(16:1)) and oleic (Delta9-cisC(18:1)) acids. The decreased transcription of the OLE1 gene (encoding the Delta9 fatty acid desaturase that catalyses the conversion of palmitic and stearic acids to palmitoleic and oleic acids, respectively) registered in 2,4-D adapted cells suggests that yeast adaptation to the herbicide involves the enhancement of the ratio of saturated (C(16:0) and C(18:0)) to monounsaturated (C(16:1) and C(18:1)) membrane fatty acids through a reduced OLE1 expression. PMID- 15781262 TI - The severe acute respiratory syndrome coronavirus 3a is a novel structural protein. AB - The severe acute respiratory syndrome coronavirus (SARS-CoV) 3a protein is one of the opening reading frames in the viral genome with no homologue in other known coronaviruses. Expression of the 3a protein has been demonstrated during both in vitro and in vivo infection. Here we present biochemical data to show that 3a is a novel coronavirus structural protein. 3a was detected in virions purified from SARS-CoV infected Vero E6 cells although two truncated products were present predominantly instead of the full-length protein. In Vero E6 cells transiently transfected with a cDNA construct for expressing 3a, a similar cleavage was observed. Furthermore, co-expression of 3a, membrane and envelope proteins using the baculovirus system showed that both full-length and truncated 3a can be assembled into virus-like particles. This is the first report that demonstrated the incorporation of 3a into virion and showed that the SARS-CoV encodes a novel coronavirus structural protein. PMID- 15781263 TI - A new human gene hNTKL-BP1 interacts with hPirh2. AB - NTKL (N-terminal kinase-like protein) encodes an evolutionarily conserved kinase like protein and is mapped around chromosomal breakpoints found in several carcinomas, suggesting that NTKL dysfunction may be involved in carcinogenesis. Recently, we identified a novel mouse gene, mNTKL-BP1 (NTKL-binding protein 1), encoding a protein interacting with NTKL. For further study, a new human gene, hNTKL-BP1, which is highly homologous with mNTKL-BP1, was used as bait in yeast two-hybrid system. hPirh2 (human p53-induced RING-H2 protein) was identified as hNTKL-BP1 interacting protein. The specific interaction of two proteins was confirmed by pull-down assay in vitro and co-immunoprecipitation in vivo. Moreover, an immunofluorescent staining assay showed that hNTKL-BP1 colocalizes with hPirh2 in SMMC 7721 cells. It will stimulate further investigation into whether hNTKL-BP1 is the substrate of hPirh2 or interaction of hNTKL-BP1 with hPirh2 enhances or represses the ubiquitin-protein ligase activity of hPirh2. PMID- 15781264 TI - Molecular analysis of a novel hereditary C3 deficiency with systemic lupus erythematosus. AB - A case of inherited homozygous complement C3 deficiency (C3D) in a patient with systemic lupus erythematosus (SLE) and the molecular basis for this deficiency are reported. A 22-year-old Japanese male was diagnosed as having SLE and his medical history revealed recurrent tonsillitis and pneumonia. He was diagnosed as having C3D because of undetectable serum C3 level. His parents were consanguineous. Sequence analysis of C3D cDNA revealed a homozygous deletion of exon 39 (84bp). A single base substitution (AG to GG) in the 3'-splice acceptor site of intron 38 was identified by sequencing the genomic DNA. Expression of C3Delta(ex39) cDNA, the C3cDNA lacking exon 39, in COS-7 cells revealed that C3Delta(ex39) was retained in endoplasmic reticulum-Golgi intermediate compartment because of defective secretion. These data indicate that a novel AG- >GG 3'-splice acceptor site mutation in intron 38 caused aberrant splicing of exon 39, resulting in defective secretion of C3. PMID- 15781265 TI - Reduced STAT3 activity in mice mimics clinical disease syndromes. AB - Phosphorylation on Y705 is obligatory for STAT3 activation, but full transcriptional activity of this widely expressed protein also requires phosphorylation on S727. We described earlier the STAT3 SA/- mice (SA, S727A allele) on a Black 6 (Bl6) background that showed 75% perinatal lethality and early growth retardation presumably due to the decreased transcription supported by STAT3 S727A. We now report additional analyses of long-term surviving SA/- animals which show no important tissue abnormalities. However, we have found a much greater susceptibility to doxorubicin-induced heart failure in the SA/- mice. Also we introduced the SA allele into strain 129 and found the SA/- mice showed greater susceptibility to LPS-induced toxicity. These results suggest a continued need for normal STAT3 transcriptional activity to resist two different noxious challenges that mimic the conditions necessary to induce adult diseases. PMID- 15781266 TI - Conformational determinants of the intracellular localization of midkine. AB - Midkine (MK) is a multifunctional growth factor and has been discovered to play important roles in carcinogenesis. MK has been reported to localize to the nucleus and nucleolus, however, the data are not consistent and the signals responsible for the localization are unknown. Here we reported that human MK exclusively localized to the nucleus and nucleolus in HepG2 cells by using GFP as a tracking molecule. In order to identify the motifs required for the nuclear localization and nucleolar accumulation, point- and deletion-mutations were introduced and the corresponding subcellular localizations were analyzed. Data revealed that (i) K79R81, K86K87, and the C-terminal tail of MK constitute the nuclear localization determinant of MK, and (ii) the C-terminal tail is the key element controlling MK nucleolar accumulation though the N-terminal tail, K79R81, and K86K87 also contribute to this process. Taken together, our results provide the first documentation about the determinants required for MK nuclear and nucleolar localization. PMID- 15781267 TI - Suppression of PI3K/mTOR pathway rescues LLC cells from cell death induced by hypoxia. AB - Cancer cells in solid tumors are challenged by various microenvironmental stresses, including hypoxia, and cancer cells in hypoxic regions are resistant to current cancer therapies. To investigate the mechanism of resistance to hypoxia in cancer cells, we examined mouse Lewis lung carcinoma (LLC) cells, which died due to necrosis at high density under hypoxic but not under normoxic conditions. Levels of mammalian target of rapamycin (mTOR), a central regulator of cellular energy, are reported to be suppressed in hypoxia. We found that phosphorylation of two molecules downstream to it, ribosomal p70 S6 kinase (S6K) and ribosomal protein S6, was markedly suppressed by hypoxia. Overexpression of the active form of S6K increased the sensitivity of LLC cells to hypoxia. On the other hand, inhibition of PI3K or mTOR dramatically reduced hypoxia-induced cell death under hypoxic conditions. Under hypoxic conditions, blockade of the PI3K or mTOR pathway increased levels of intracellular ATP and delayed decreases in pH and glucose level in culture medium, without affecting the cell cycle. PMID- 15781268 TI - Glucagon production of the rat insulinoma cell line INS-1-A quantitative comparison with primary rat pancreatic islets. AB - The rat insulinoma cell line INS-1 is the most commonly used clonal cell model in pancreatic beta-cell research. Considering the multihormonality of many insulinomas we examined as to how INS-1 cells comply with the notion of resembling a pure beta-cell line. Glucagon immunoassays revealed that INS-1 cells secrete glucagon in a similar range as islets. By immunohistochemistry we detected a cytoplasmic glucagon signal in INS-1 cells which colocalized with C peptide. Cellular content of preproglucagon-mRNA and glucagon protein in INS-1 cells was less than two percent of the respective values in islets, which probably reflects differences in the intracellular metabolism and/or secretory pathways. Taken together, it is obvious that INS-1 cells do not represent an exclusively insulin producing beta-cell line. PMID- 15781269 TI - The RET finger protein interacts with the hinge region of SMC3. AB - The structural maintenance of chromosome 3 protein (SMC3) is a component of the multimeric cohesin complex that holds sister chromatids together and prevents their premature separation during mitosis. By screening a human cDNA library for interacting proteins we have established that the proto-oncogene RET finger protein (RFP) interacts with SMC3. The sites of interaction map to part of the central coiled coil region of RFP and to the C-terminal region of the SMC3 globular hinge domain. SMC3/RFP interaction was confirmed in vivo by co immunoprecipitation studies and by performing mammalian two-hybrid interaction assays. Cytoimmunolocalization experiments showed that SMC3 and RFP co-localize in the same cell substructures. Overexpression of RFP in NIH3T3 cells significantly increased the fraction of SMC3 recovered in the nucleus supporting the idea that RFP regulates the intracellular distribution of SMC3. These studies identify a novel SMC3-interacting protein that may affect SMC3 availability to complex with its cohesin partners. PMID- 15781270 TI - The effect of cytoskeletal disruption on pulsatile fluid flow-induced nitric oxide and prostaglandin E2 release in osteocytes and osteoblasts. AB - Fluid flowing through the bone porosity might be a primary stimulus for functional adaptation of bone. Osteoblasts, and osteocytes in particular, respond to fluid flow in vitro with enhanced nitric oxide (NO) and prostaglandin E(2) (PGE(2)) release; both of these signaling molecules mediate mechanically-induced bone formation. Because the cell cytoskeleton is involved in signal transduction, we hypothesized that the pulsatile fluid flow-induced release of NO and PGE(2) in both osteoblastic and osteocytic cells involves the actin and microtubule cytoskeleton. In testing this hypothesis we found that fluid flow-induced NO response in osteoblasts was accompanied by parallel alignment of stress fibers, whereas PGE(2) response was related to fluid flow stimulation of focal adhesions formed after cytoskeletal disruption. Fluid flow-induced PGE(2) response in osteocytes was inhibited by cytoskeletal disruption, whereas in osteoblasts it was enhanced. These opposite PGE(2) responses are likely related to differences in cytoskeletal composition (osteocyte structure was more dependent on actin), but may occur via cytoskeletal modulation of shear/stretch-sensitive ion channels that are known to be dominant in osteocyte (and not osteoblast) response to mechanical loading. PMID- 15781271 TI - The overexpression of an intracellular ABCA-like transporter alters phospholipid trafficking in Leishmania. AB - In parasites, ATP-binding cassette (ABC) transporters represent an important family of proteins related to drug resistance and other biological activities. Here we report the characterization of LtrABCA2, a new ABC transporter of the ABCA subfamily in the protozoan parasite Leishmania tropica, localized at the flagellar pocket region and in internal vesicles. The overexpression of this transporter reduced the accumulation of fluorescent glycerophospholipid analogs, increased the exocytic activity, and decreased infectivity of macrophage, but did not confer resistance to drugs. Together, these results suggest that this new ABC transporter plays a role in phospholipid trafficking, which may be modifying the vesicular trafficking and the infectivity of the parasite. PMID- 15781273 TI - Cellular and molecular events in corneal wound healing: significance of lipid signalling. AB - Alterations in the normal healing process after corneal injury can produce undesirable outcomes that range from corneal haze to ulceration and perforation. Lipids play important roles in the complex inflammatory processes that occur after corneal wounding. While some lipid mediators, such as the lipoxygenase derivatives of arachidonic acid, 12-hydroxyeicosatetraenoic acid (12[S]-HETE and 15[S]-HETE), act as second messengers to promote cell proliferation and are possibly involved in the synthesis of other molecules that suppress inflammation, others, such as platelet-activating factor (PAF), exert their actions through specific receptors, play key roles during sustained corneal inflammation (as might occur with chemical burns), and contribute to tissue destruction and neovascularization. PAF is also a strong inducer of selective metalloproteinases (MMPs) that degrade the extracellular matrix. The use of a new PAF antagonist has shown great promise for the treatment of diffuse lamellar keratitis (DLK) and alkali-burned corneas. PMID- 15781274 TI - Potentiation of ATP-induced Ca2+ mobilisation in human retinal pigment epithelial cells. AB - Interaction of signalling pathways directs the functional output of many cells. This study investigated the consequences of activating adenosine and adrenergic receptors on ATP-induced Ca2+ responses in human retinal pigment epithelial (RPE) cells. Intracellular Ca2+ concentration ([Ca2+]i) of human RPE cells in primary culture was monitored using Fura-2. Cyclic adenosine monophosphate (cAMP) concentration was measured using an enzyme-linked immunosorbent assay. Both ATP and UTP (10 microM) increased [Ca2+]i in human RPE cells. Adenosine (10 nM-10 microM) had no effect on resting [Ca2+]i, but potentiated a sub-threshold response to ATP (100 nM) when ATP was added in the presence of adenosine. The potentiation occurred with other G-protein receptor agonists such as acetylcholine. Potentiation persisted in Ca-free medium, but was blocked by prior application of thapsigargin. The A1 and A2 adenosine receptor antagonists, DPCPX and MRS1706 (100 nM) respectively, inhibited potentiation in 76+/-7 and 23+/-12% of cells, respectively, but the A3 antagonist MRS1191 had no effect. Conversely, agents that activate the cAMP pathway, including isoproterenol (10 microM), forskolin (10 microM), and the protein kinase A (PKA) activator Sp-cBIMPS (1 microM), potentiated the ATP-induced response in the RPE cells. Agents that are known to inhibit the production of cAMP in other systems also caused potentiation, including clonidine (10 microM) and the Gi-activator mastoparan (10 microM). Under resting conditions, cAMP concentration in RPE cells was 7.1+/-0.5 pmol mg(-1) protein. Isoproterenol (10 microM) and forskolin (10 microM) increased levels to 104.6+/-5.2 and 113.7+/-4.2 pmol mg(-1) protein, respectively, while adenosine, clonidine, and mastoparan (all 10 microM) had no significant effect on cAMP levels. These data indicate that whilst activation of A1 and A2 adenosine receptors and alpha2 and beta adrenergic receptors does not influence basal Ca2+ levels, stimulation of these receptors can potentiate Ca2+ signalling by cAMP dependent and independent mechanisms in human RPE cells. PMID- 15781276 TI - Adenosine agonist regulation of outward active transport of fluorescein across retinal pigment epithelium in rabbits. AB - To investigate the effect of an adenosine agonist, 2-5'-N ethylcarboxamidoadenosine (NECA), on the outward active transport of fluorescein across the retinal pigment epithelium (RPE) in rabbits. High (5x10(-4)-2x10(-3) M) and low (1x10(-5)-1x10(-4) M) concentrations of NECA or phosphate buffered saline (PBS) were intravitreously injected into Dutch-belted rabbits. Sodium fluorescein was injected intravenously 180 min after NECA. Differential vitreous fluorophotometry was performed 3 hr after the sodium fluorescein injection and the vitreal fluorescein/fluorescein monoglucuronide (F/FG) ratio then was calculated. The F/FG ratios are inversely proportional to the outward active transport of fluorescein across the RPE. Retinal detachments were induced by injection of PBS into the subretinal space after the intravitreous injection of low- or high-dose NECA or PBS, and the size of the blebs was monitored. In eyes that received a low-dose injection of NECA, the F/FG ratio was higher compared with controls (P<0.05); in eyes that received a high-dose intravitreal injection, the F/FG ratio was significantly lower compared with controls (P<0.05). The effect of low-dose NECA on the F/FG ratio was suppressed by the A2 receptor antagonist, ZM241385, and the effect of high-dose NECA was suppressed by the A1 receptor antagonist, 8-cyclopentyl-1, 3-dipropylxanthine. The A3 receptor antagonist MRS1191 did not influence the effect of low- or high-dose NECA. Intravitreal injection of high-dose NECA enhanced the reabsorption of subretinal fluid compared with PBS; however, low-dose NECA inhibited reabsorption of subretinal fluid (P<0.02 and 0.05, respectively). Intravitreous injection of high dose NECA accelerates the active outward transport across the RPE via A1 receptors and low-dose NECA decelerates it via A2 receptors. PMID- 15781278 TI - Immunohistochemical detection and Western blot analysis of nitrated protein in stored human corneal epithelium. AB - While the production of nitric oxide by human corneas in storage has recently been demonstrated, protein nitration as a result of this production has not been demonstrated. In this study, nitrated protein accumulation in the epithelium of stored human corneas was assessed. One half of five donor corneas maintained in storage media for 3 days were prepared for immunohistochemical studies. The other halves remained in storage media for 7 additional days and were also processed for immunohistochemistry. Mouse monoclonal antibody to nitrotyrosine adducts was used to define the localisation of these epitopes. The density of antibody staining was observed and quantified on a digital camera system and statistically analysed. Immunostaining in the epithelium was greater in tissues recovered after 10 days in storage compared to the intensity of staining after 3 days of storage (p<0.0001). No staining was evident in the epithelium in sections exposed to non immune mouse IgG. Western blot analysis was performed on epithelial cells scraped from corneal surfaces of one-half of four donor corneas in storage for 3 days and from the other half at 10 days of storage. Nitrated BSA was used as a positive control. After extraction and homogenisation, identical protein concentrations of each sample were loaded per lane on 10% gels and subjected to SDS-PAGE. Proteins were blotted and probed with the anti-nitrotyrosine antibody. Western blot immunoreactivity was detected in epithelial samples at the 3 and 10 day recovery times with the latter samples showing greater staining intensity. Nitrated protein, thought to indicate toxic peroxynitrite formation, accumulates in the human corneal epithelium with time of storage. Our study shows that there is an association between increased nitrated protein and storage time. PMID- 15781277 TI - Cytoprotection by nipradilol, an anti-glaucomatous agent, via down-regulation of apoptosis related gene expression and activation of NF-kappaB. AB - It has been reported that nipradilol, a nonselective beta- and selective alpha1 receptor antagonist, has cytoprotective effects. We attempted to clarify the effects of nipradilol on the expression of apoptosis associated genes and the activity of nuclear factor-kappaB, a transcription factor, in PC12 cells during serum deprivation induced apoptosis. PC12 cells were cultured in serum free RPMI1640 medium with or without 0.01, 0.1, 1, or 10 microM of nipradilol, or in serum-added medium as a control. The gene expressions of Bax, Bcl-2, Fas, FasL, Caspase-1, 2, 3, and 9, p53, and Smac/DIABLO were examined using a quantitative real time polymerase chain reaction method, while nuclear factor-kappaB activity was examined using an electrophoresis mobility shift assay with a nuclear factor kappaB consensus sequenced DNA probe. The effects of denitronipradilol were also examined to clarify the effect of nitric oxide donative action. Nipradilol down regulated Bax gene expression 12 hr after serum deprivation, and that of the capase-9 and Smac/DIABLO genes at 24 hr, compared to the serum-free sample, while it also increased cell viability and decreased DNA ladder formation at 48 hr. However, the expressions of other examined genes were not affected by the agent. In addition, nuclear factor-kappaB activity was increased 2 hr after the addition of 0.1 or 1 microM of nipradilol. In contrast, denitronipradilol did not show any effects toward PC12 cells. Our results suggest that nipradilol may have an effect on apoptosis associated gene expression and nuclear factor-kappaB activity during the prevention of apoptosis via nitric oxide donative action. PMID- 15781275 TI - Age-dependent alterations in mouse exorbital lacrimal gland structure, innervation and secretory response. AB - Several studies investigated the effect of aging on rat and human lacrimal gland physiology. However, in most of these studies, only two age groups were investigated. Furthermore, those studies did not correlate the age-related histological changes that occur in the lacrimal gland to the functional changes (nerve activity and protein secretion) that might occur with aging. Thus, the purpose of the present study was to investigate the effect of aging on lacrimal gland structure, innervation and function using BALB/c mice at different ages. Exorbital lacrimal glands were removed from 3, 8, 12, 24, and 32-month-old, male BALB/c mice, fixed, embedded and processed for histology and immunohistochemistry. Sections were stained with hematoxylin and eosin to determine morphological changes and lymphocytic infiltration; giemsa to identify mast cells; and Kinyoun's carbol fucsin solution to indicate lipofuscin-like inclusions. Parasympathetic and sympathetic nerves were identified by immunofluorescence techniques. To measure acetylcholine release and protein secretion, lacrimal gland pieces were incubated in Krebs Ringer buffer containing 5 mM KCl (control), 75 mM KCl (depolarizing buffer which activates nerves), carbachol (a cholinergic agonist, 10(-4) M), or phenylephrine (an alpha1 adrenergic agonist, 10(-4) M) for 20 min. The media were collected and analysed for acetylcholine and peroxidase using a spectrofluorometric assay. KCl-, carbachol- and phenylephrine-stimulated peroxidase secretion decreased in lacrimal glands from 8, 12, and 24-month-old mice when compared to 3-month-old animals. Both the density and distribution of parasympathetic and sympathetic nerves surrounding the acini decreased with increasing age. Acetylcholine release from lacrimal gland nerves decreased in 24-month-old mice compared to 3- and 12 month-old animals. Similarly, progressive morphological changes, including increased numbers of lipofuscin-like inclusions, mast cells and lymphocytic infiltration occurred in an age-dependent manner. We conclude that structural alterations of mouse lacrimal gland, including increased accumulation of lipofuscin-like inclusions, chronic inflammation and functional alterations including decreased acetylcholine release and protein secretion occurred with aging. PMID- 15781279 TI - Photoreceptor differentiation and integration of retinal progenitor cells transplanted into transgenic rats. AB - Previous studies evaluating neural stem cells transplanted into the mature retina have demonstrated limited levels of graft-host integration and photoreceptor differentiation. The purpose of this investigation is to enhance photoreceptor cell differentiation and integration of retinal progenitor cells (RPC) following subretinal transplantation into retinal degenerate rats by optimization of isolation, expansion, and transplantation procedures. RPCs were isolated from human placental alkaline phosphatase (hPAP)-positive embryonic day 17 (E17) rat retina and expanded in serum-free defined media. RPCs at passage 2 underwent in vitro induction with all trans retinoic acid or were transplanted into the subretinal space of post-natal day (P) 17 S334ter-3 and S334ter-5 transgenic rats. Animals were examined post-operatively by ophthalmoscopy and optical coherence tomography (OCT) at weeks 1 and 4. Differentiation profiles of RPCs, both in vitro and in vivo were analysed microscopically by immunohistochemistry for various retinal cell specific markers. Our results demonstrated that the majority of passage 2 RPCs differentiated into retina-specific neurons expressing rhodopsin after in vitro induction. Following subretinal transplantation, grafted cells formed a multi-layer cellular sheet in the subretinal space in both S334ter 3 and S334ter-5 rats. Prominent retina-specific neuronal differentiation was observed in both rat lines as evidenced by recoverin or rhodopsin staining in 80% of grafted cells. Less than 5% of the grafted cells expressed glial fibrillary acidic protein. Synapsin-1 (label for nerve terminals) positive neural processes were present at the graft-host interface. Expression profiles of the grafted RPCs were similar to those of RPCs induced to differentiate in vitro using all-trans retinoic acid. In contrast to our previous study, grafted RPCs can demonstrate extensive rhodopsin expression, organize into layers, and show some features of apparent integration with the host retina following subretinal transplantation in slow and fast retinal degenerate rats. The similarity of the in vitro and in vivo RPC differentiation profiles suggests that intrinsic signals may have a significant contribution to RPC cell fate determination. PMID- 15781281 TI - The protective role of squalene in alcohol damage in the chick embryo retina. AB - The developing CNS, and in particular the visual system, is very sensitive to the effects of alcohol. Alcohol causes lipid peroxidation. Squalene, the major olive oil hydrocarbon, is a quencher of singlet oxygen and prevents the corresponding lipid peroxidation. We presumed that squalene can protect against the alcohol induced damage already observed during the development of the chick retina. Alcohol+squalene was administered directly into the yolk sac of the egg of White Leghorn chicks at day 6 of incubation. The lipid composition of the retina was analyzed in embryos at E7, E11, E15 and E18. The proportions of phospholipids, free and esterified cholesterol, diacylglycerides and free fatty acids were estimated using the Iatroscan TLC/FID procedure. Gas chromatography and mass spectrometry were used to determine the fatty acid composition. The morphological study was carried out at E11 using semithin sections, and by means of immunohistochemical techniques at E19. Comparing the results obtained in control embryos, the administration of alcohol+squalene reduces the effects of alcohol on the total lipid composition of the retina during development. The effects were, in fact, of less magnitude than in embryos treated only with alcohol. The major phospholipid species of alcohol+squalene-treated embryos exhibited total recuperation at E15. As far as fatty acids are concerned, no significant changes were observed with regard to control embryos during development. From a morphological point of view, the retinas of alcohol+squalene-treated embryos show at E11 fewer cellular alterations than the retinas of alcohol-treated embryos. In this respect, the retinas of alcohol+squalene-treated embryos exhibited: a columnar cell arrangement similar to that observed in control retinas; few pycnotic cells and very few alterations in ganglion cell layers and in the optic nerve fibers layer. At E19 the recuperation of the expression of myelin oligodendrocyte specific protein (MOSP) in alcohol+squalene-treated embryos was recorded. Since squalene reduces the deleterious effects caused by alcohol on the lipid composition and the structure of the retina, squalene could act as a naturally occurring agent for the prevention of damage caused by abusive alcohol ingestion during pregnancy. PMID- 15781280 TI - Retinal arterial occlusion leads to acidosis in the cat. AB - This study investigated the changes in pH during retinal artery occlusion by means of extracellular H+ concentration ([H+]o) measurements in the retina under both air and 100% O2 ventilation. Occlusion was produced in intact anesthetised cats by pressing with a probe onto a retinal artery. [H+]o profiles were recorded across the retina with pH sensitive microelectrodes. The average inner retinal [H+]o increased during occlusion, resulting in an acidification of as much as 0.10 pH units, even under 100% O2 ventilation. The inner retinal H+ profile magnitude decreased during occlusion due to impaired clearance. The average outer retinal H+ profile magnitude also increased even though outer retinal H+ production did not increase during occlusion. This might be due to H+ diffusion from the inner retina to the outer retina, which is opposite to the flux in the normal retina. After reperfusion, [H+]o returned to its preocclusion value. In conclusion, arterial occlusion leads to acidification of the retina. Enhanced oxygenation during occlusion did not decrease this acidification. This may explain why increasing PO2 in the retina by enhanced O2 breathing improves retinal function during and after occlusion, but does not totally reverse the effect of occlusion. PMID- 15781282 TI - Conjunctival M cells selectively bind and translocate Maackia amurensis leukoagglutinin. AB - Antigen-sampling M cells are found in the follicle-associated epithelium above organized lymphoid tissue in many mucosae. They play a key role in initiating the mucosal immune response and act as a site of entry for opportunistic pathogens. This study investigates the presence of M cells in the Guinea pig conjunctiva. Maackia amurensis leukoagglutinin I and II (MAL-I and MAL-II) were identified as potential conjunctival M cell markers based on a screening of 12 lectins and 5 carbohydrate epitope antibodies on aldehyde-fixed follicles. Biotinylated or fluorescein-conjugated MAL-I was then instilled into conjunctival sacs in vivo for 15-60 min. Specimens were assessed by epi-fluorescence stereomicroscopy, confocal scanning laser microscopy and transmission and scanning electron microscopy (TEM and SEM). Selective labelling of a subset of epithelial cells overlying lymphoid follicles was observed following in vivo exposure to MAL-I. MAL-I labelling was restricted to cells with sparse, irregular microvilli. Cells preferentially labelled with MAL-I were found to internalize the lectin during a 60 min in vivo exposure. MAL-I was transcytosed to basolateral membranes of cells filled with intracellular vesicles during a 45 min in vivo incubation. This study demonstrates that the Guinea pig conjunctiva contains a cell with morphological and functional characteristics of antigen-sampling M cells. PMID- 15781283 TI - TGF-beta1, IL-1beta, and Th2 cytokines stimulate vascular endothelial growth factor production from conjunctival fibroblasts. AB - Giant papillary formation containing newly formed vessels is a major characteristic of severe allergic conjunctivitis, such as atopic keratoconjunctivitis (AKC) or vernal keratoconjunctivitis (VKC). We examined production of vascular endothelial growth factor (VEGF) from cultured conjunctival fibroblasts from normal volunteers under stimulation with type 1-, type 2-helper T cell derived and proinflammatory cytokines to investigate the mechanism of giant papillae formation in AKC/VKC. Primary cultured conjunctival fibroblasts were incubated with interleukin (IL)-4, IL-13, IL-1beta, IL-2, tumor necrotizing factor (TNF)-alpha, interferon (IFN)-gamma, or transforming growth factor (TGF)-beta1. Effects of cytokines on VEGF protein secretion in supernatant were assessed by ELISA, and VEGF mRNA expression in cultured cells were assessed by quantitative PCR. TGF-beta1 most effectively increased VEGF concentration with dose- and time-dependent manner IL-1beta, IL-4, and IL-13 significantly increased VEGF concentration. Though IL-2 also showed slight increase of VEGF concentration, it was not statistically significant. TNF-alpha and INF-gamma did not increase VEGF concentration. Quantitative PCR showed significant increase of VEGF mRNA in TGF-beta1, IL-1beta, and IL-4 stimulated fibroblasts. TGF-beta1, IL 1beta, and Th2 cytokines from allergic inflammatory cells induced VEGF production in conjunctival fibroblasts, and may play a crucial role in neovascularization and formation of giant papillae in AKC/VKC. PMID- 15781284 TI - Maximum tolerable dose for avoidance of cataract induced by ultraviolet radiation B for 18 to 60 week old rats. AB - The purpose of the present study was to investigate the maximum tolerable dose for avoidance of UVR-B-induced cataract in rats in the age interval 18-60 weeks and establish the functional relationship between age and sensitivity to UVR-B. Four groups of 20 albino Sprague-Dawley rats each, aged 18, 26, 40 or 60 weeks, were included. Each age group was divided into five UVR dose sub-groups. The rats were unilaterally exposed to ultraviolet radiation (lambda(max)=302.6 nm, lambda0.5=4.5 nm). The incident dose on the cornea varied between 0 and 9.2 kJ m( 2). One week after exposure, the rats were sacrificed and both lenses were extracted. The intensity of forward light scattering was measured and photographs were taken. The functional relationship between age and sensitivity to UVR-B was estimated as the maximum tolerable dose based on rats age from 3 to 60 weeks. The maximum tolerable dose for 18, 26, 40, and 60 weeks, respectively, was estimated to 5.2, 4.9, 4.7, and 5.1 kJ m(-2). The sensitivity to UVR-B for Sprague-Dawley rats increases with increasing age during the first third of the rat life span, and then stabilizes to a constant level during the remaining two-thirds. PMID- 15781285 TI - Mechanisms for the induction of HNE- MDA- and AGE-adducts, RAGE and VEGF in retinal pigment epithelial cells. AB - Pathological features of age-related macular degeneration such as the formation of extracellular deposits and neovascularization are frequently viewed as outcomes of compromising processes within retinal pigment epithelial cells, but the initiating circumstances are poorly understood. Here we tested the hypothesis that photooxidation events initiated by A2E, a blue light-excitable aging fluorophore of the retinal pigment epithelium, can set the stage for altered cellular signaling and changes in the expression of genes that can impact the extracellular milieu. Proteins modified by lipid peroxidation products (4 hydroxynonenal; malondialdhyde) and advanced glycation end products were detected at sites of blue light irradiation both in association with the cultured A2E laden retinal pigment epithelial cells and within the fibronectin substrate on which the cells were grown. RAGE, the cell surface receptor that transduces the effects of advanced glycation end products, was also upregulated, and RAGE expression co-localized with the deposition of advanced glycation end products. Blue light triggered alterations in gene expression was also evidenced by elevations in both transcripts and protein for vascular endothelial growth factor, a potent angiogenic and permeability-enhancing factor. These findings indicate that cell associated and extracellular modification of proteins by lipid peroxidation products and advanced glycation end products together with increased expression of RAGE and vascular endothelial growth factor may be induced consequent to blue light illumination of A2E-burdened retinal pigment epithelial cells. Thus, photooxidative events that are not an immediate threat to retinal pigment epithelial cell viability may nevertheless elicit sustained perturbation that could ultimately alter neighboring tissues and impact retinal pigment epithelial cell function. PMID- 15781287 TI - A simple modification significantly reduces a background staining in immunostaining of corneal tissues using a biotinylated rabbit polyclonal antibody. PMID- 15781286 TI - Partial enrichment of a population of human limbal epithelial cells with putative stem cell properties based on collagen type IV adhesiveness. AB - The concept that corneal epithelium stem cells reside in limbus has been recognized for more than a decade, but isolation of these stem cells has not been accomplished. This study was an initial attempt to isolate a population of human limbal epithelial cells enriched for certain putative stem cell properties based on their phenotype. Epithelial cells harvested from fresh human limbal rings and their primary cultures were allowed to adhere to collagen IV-coated dishes for 20 min and 2 hr, sequentially. The rapidly adherent cells (RAC), slowly adherent cells and non-adherent cells were evaluated for certain stem cell properties: (a) BrdU-label retention, (b) expression of basal cell (integrin beta1, p63, ABCG2) and differentiation (involucrin, keratin 12) markers, and (c) colony forming efficiency (CFE) and growth capacity on a 3T3 fibroblast feeder layer. Among unfractionated cells and the three selected populations, the RAC, accounting for about 10% of whole population, were enriched 5-fold in BrdU label-retaining cells, displayed the highest number of integrin beta1 and p63 positive and involucrin negative cells, expressed high levels of DeltaNp63 and ABCG2 mRNA, and lacked involucrin and K12 expression, and possessed the greatest CFE and growth capacity. These findings demonstrated for the first time that human limbal epithelial cells with stem cell properties can be partially enriched by their adhesiveness to collagen IV. The RAC population enriched for certain putative stem cell properties may prove useful in the future for transplantation to diseased and damaged corneas with limbal stem cell deficiency. PMID- 15781288 TI - Toxicological awakenings: the rebirth of hormesis as a central pillar of toxicology. AB - This paper assesses historical reasons that may account for the marginalization of hormesis as a dose-response model in the biomedical sciences in general and toxicology in particular. The most significant and enduring explanatory factors are the early and close association of the concept of hormesis with the highly controversial medical practice of homeopathy and the difficulty in assessing hormesis with high-dose testing protocols which have dominated the discipline of toxicology, especially regulatory toxicology. The long-standing and intensely acrimonious conflict between homeopathy and "traditional" medicine (allopathy) lead to the exclusion of the hormesis concept from a vast array of medical- and public health-related activities including research, teaching, grant funding, publishing, professional societal meetings, and regulatory initiatives of governmental agencies and their advisory bodies. Recent publications indicate that the hormetic dose-response is far more common and fundamental than the dose response models [threshold/linear no threshold (LNT)] used in toxicology and risk assessment, and by governmental regulatory agencies in the establishment of exposure standards for workers and the general public. Acceptance of the possibility of hormesis has the potential to profoundly affect the practice of toxicology and risk assessment, especially with respect to carcinogen assessment. PMID- 15781289 TI - Induction of hepatic and renal metallothionein synthesis by ferric nitrilotriacetate in mice: the role of MT as an antioxidant. AB - Metallothionein (MT) demonstrates strong antioxidant properties, yet the physiological relevance of its antioxidant action is not clear. Injection of mice with ferric nitrilotriacetate (Fe-NTA) caused a dose-dependent increase in hepatic and renal MT. Fe-NTA caused a greater increase in hepatic and renal MT concentration (2.5- and 4-fold) compared with FeCl(3) at the same dose of ferric ion. MT mRNA levels were markedly elevated in both of tissues. Thiobarbituric acid (TBA) values in both tissues reached a maximum after 2-4 h. The MT concentrations were significantly increased after 2-4 h in liver and after 8-16 h in kidneys. Plasma concentrations of cytokines such as IL-6 and TNFalpha were elevated by 4 h; IL-6 levels were 24 times higher after Fe-NTA than that after injection of FeCl(3). Pretreatment of mice with ZnSO(4) attenuated nephrotoxicity induced by Fe-NTA after 2 h, but was not effective 4 h after injection. After a Fe-NTA injection, a loss of Cd-binding properties of preinduced MT was observed only in kidneys of Zn-pretreated mice but not in liver. Treatment with BSO, glutathione (GSH) depletor, intensified a loss of its Cd-binding properties after a Fe-NTA injection. These results indicate that induction of MT synthesis may result from reactive oxygen species (ROS) generated by Fe-NTA, and MT may act in vivo as a complementary antioxidant. PMID- 15781290 TI - Implications of oxidative stress and hepatic cytokine (TNF-alpha and IL-6) response in the pathogenesis of hepatic collagenesis in chronic arsenic toxicity. AB - INTRODUCTION: Noncirrhotic portal fibrosis has been reported to occur in humans due to prolonged intake of arsenic contaminated water. Further, oxystress and hepatic fibrosis have been demonstrated by us in chronic arsenic induced hepatic damage in murine model. Cytokines like tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) are suspected to play a role in hepatic collagenesis. The present study has been carried out to find out whether increased oxystress and cytokine response are associated with increased accumulation of collagen in the liver due to prolonged arsenic exposure and these follow a dose-response relationship. METHODS: Male BALB/c mice were given orally 200 microl of water containing arsenic in a dose of 50, 100, and 150 mug/mouse/day for 6 days a week (experimental group) or arsenic-free water (<0.01 microg/l, control group) for 3, 6, 9 and 12 months. Hepatic glutathione (GSH), protein sulfhydryl (PSH), glutathione peroxidase (GPx), Catalase, lipid peroxidation (LPx), protein carbonyl (PC), interleukin (IL-6), tumor necrosis factor (TNF-alpha), arsenic and collagen content in the liver were estimated from sacrificed animals. RESULTS: Significant increase of lipid peroxidation and protein oxidation in the liver associated with depletion of hepatic thiols (GSH, PSH), and antioxidant enzymes (GPx, Catalase) occurred in mice due to prolonged arsenic exposure in a dose dependent manner. Significant elevation of hepatic collagen occurred at 9 and 12 months in all the groups associated with significant elevation of TNF-alpha and IL-6. However, arsenic level in the liver increased progressively from 3 months onwards. There was a positive correlation between the hepatic arsenic level and collagen content (r = 0.8007), LPx (r = 0.779) and IL-6 (r = 0.7801). Further, there was a significant negative correlation between GSH and TNF-alpha (r = 0.5336)) and LPx (r = -0.644). CONCLUSION: Increasing dose and duration of arsenic exposure in mice cause progressive increase of oxystress and elevation of cytokines associated with increasing level of collagen in the liver. PMID- 15781291 TI - Suppressive effects of ketamine on macrophage functions. AB - Ketamine is an intravenous anesthetic agent. Clinically, induction of anesthesia with ketamine can cause immunosuppression. Macrophages play important roles in host defense. In this study, we attempted to evaluate the effects of ketamine on macrophage functions and its possible mechanism using mouse macrophage-like Raw 264.7 cells as the experimental model. Exposure of macrophages to 10 and 100 microM ketamine, which correspond to 0.1 and 1 times the clinically relevant concentration, for 1, 6, and 24 h had no effect on cell viability or lactate dehydrogenase release. When the administered concentration reached 1000 microM, ketamine caused a release of lactate dehydrogenase and cell death. Ketamine, at 10 and 100 microM, did not affect the chemotactic activity of macrophages. Administration of 1000 microM ketamine in macrophages resulted in a decrease in cell migration. Treatment of macrophages with ketamine reduced phagocytic activities. The oxidative ability of macrophages was suppressed by ketamine. Treatment with lipopolysaccharide induced TNF-alpha, IL-1beta, and IL-6 mRNA in macrophages. Administration of ketamine alone did not influence TNF-alpha, IL 1beta, or IL-6 mRNA production. Meanwhile, cotreatment with ketamine and lipopolysaccharide significantly inhibited lipopolysaccharide-induced TNF-alpha, IL-1beta, and IL-6 mRNA levels. Exposure to ketamine led to a decrease in the mitochondrial membrane potential. However, the activity of mitochondrial complex I NADH dehydrogenase was not affected by ketamine. This study shows that a clinically relevant concentration of ketamine (100 microM) can suppress macrophage function of phagocytosis, its oxidative ability, and inflammatory cytokine production possibly via reduction of the mitochondrial membrane potential instead of direct cellular toxicity. PMID- 15781292 TI - Microglia is activated by astrocytes in trimethyltin intoxication. AB - Microglia participates in most acute and chronic neuropathologies and its activation appears to involve interactions with neurons and other glial cells. Trimethyltin (TMT)-induced brain damage is a well-characterized model of neurodegeneration, in which microglial activation occurs before neuronal degeneration. The aim of this in vitro study was to investigate the role of astroglia in TMT-induced microgliosis by using nitric oxide (NO), inducible NO synthase (iNOS), and morphological changes as parameters for microglial activation. Our investigation discusses (a) whether microglial cells can be activated directly by TMT; (b) if astroglial cells are capable of triggering or modulating microglial activation; (c) how the morphology and survival of microglia and astrocytes are affected by TMT treatment; and (d) whether microglial-astroglial interactions depend on direct cell contact or on soluble factors. Our results show that microglia are more vulnerable to TMT than astrocytes are and cannot be activated directly by TMT with regard to the examined parameters. In bilayer coculture with viable astroglial cells, microglia produce NO in significant amounts at subcytotoxic concentrations of TMT (20 micromol/l). At these TMT concentrations, microglial cells in coculture convert into small round cells without cell processes, whereas flat, fibroblast-like astrocytes convert into thin process bearing stellate cells with a dense and compact cell body. We conclude that astrocytes trigger microglial activation after treatment with TMT, although the mechanisms of this interaction remain unknown. PMID- 15781293 TI - Acrylonitrile potentiates hearing loss and cochlear damage induced by moderate noise exposure in rats. AB - The diversity of chemical and drugs that can potentiate noise-induced hearing loss (NIHL) has impeded efforts to predict such interactions. We have hypothesized that chemical contaminants that disrupt intrinsic antioxidant defenses hold significant risk for potentiating NIHL. If this is true, then acrylonitrile (ACN) would be expected to potentiate NIHL. ACN, one of the 50 most commonly used chemicals in the United States, is metabolized via two pathways that are likely to disrupt intrinsic reactive oxygen species (ROS) buffering systems: (1) it conjugates glutathione, depleting this important antioxidant rapidly; (2) a second pathway involves the formation of cyanide, which can inhibit superoxide dismutase. We hypothesized that moderate noise exposure, that does not produce permanent hearing loss by itself, could initiate oxidative stress and that ACN could render the inner ear more sensitive to noise by disrupting intrinsic antioxidant defenses. Temporary and persistent effects of ACN alone (50 mg/kg, sc 5 days), noise alone (95 or 97 dB octave band noise, 4 h/day for 5 days), or ACN in combination with noise were determined using distortion product otoacoustic emissions (DPOAEs) and compound action potential (CAP) amplitudes. Histopathological damage to hair cells resulting from these treatments was also investigated using surface preparations of the organ of Corti. Individually, neither ACN nor noise exposures caused any permanent hearing or hair cell loss; only a reversible temporary threshold shift was measured in noise-exposed animals. However, when given in combination, ACN and noise induced permanent threshold shifts (13-16 dB between 7 and 40 kHz) and a decrease in DPOAE amplitudes (up to 25 dB at 19 kHz), as well as significant outer hair cell (OHC) loss (up to 20% in the first row between 13 and 47 kHz). This investigation demonstrates that ACN can potentiate NIHL at noise levels that are realistic in terms of human exposure, and that the OHCs are the main target of toxicity. While the exact mechanism is unknown, the results are consistent with the hypothesis of ROS involvement in NIHL at moderate levels. PMID- 15781294 TI - Free radical production induced by methamphetamine in rat striatal synaptosomes. AB - The pro-oxidative effect of methamphetamine (METH) in dopamine terminals was studied in rat striatal synaptosomes. Flow cytometry analysis showed increased production of reactive oxygen species (ROS) in METH-treated synaptosomes, without reduction in the density of dopamine transporters. In synaptosomes from dopamine (DA)-depleted animals, METH did not induce ROS production. Reserpine, in vitro, completely inhibited METH-induced ROS production. These results point to endogenous DA as the main source of ROS induced by METH. Antioxidants and inhibitors of neuronal nitric oxide synthase and protein kinase C (PKC) prevented the METH-induced oxidative effect. EGTA and the specific antagonist methyllycaconitine (MLA, 50 microM) prevented METH-induced ROS production, thus implicating calcium and alpha7 nicotinic receptors in such effect. Higher concentrations of MLA (>100 microM) showed nonspecific antioxidant effect. Preincubation of synaptosomes with METH (1 microM) for 30 min reduced [(3)H]DA uptake by 0%. The METH effect was attenuated by MLA and EGTA and potentiated by nicotine, indicating that activation of alpha(7) nicotinic receptors and Ca(2+) entry are necessary and take place before DAT inhibition. From these findings, it can be postulated that, in our model, METH induces DA release from synaptic vesicles to the cytosol. Simultaneously, METH activates alpha(7) nicotinic receptors, probably inducing depolarization and an increase in intrasynaptosomal Ca(2+). This would lead to DAT inhibition and NOS and PKC activation, initiating oxidation of cytosolic DA. PMID- 15781295 TI - In vitro effects of diethylstilbestrol, genistein, 4-tert-butylphenol, and 4-tert octylphenol on steroidogenic activity of isolated immature rat ovarian follicles. AB - Isolated rat ovarian follicles grow and produce steroid hormones in vitro and so provide a good model for studying the effects of hormonally active compounds on follicular steroidogenesis. We have evaluated the effects of diethylstilbestrol (DES), genistein (GEN) and two alkylphenols, 4-tert-butylphenol (BP) and 4-tert octylphenol (OP) on the growth, survival, and steroid hormone and cAMP production by isolated 14-day-old rat (Sprague-Dawley) ovarian follicles. During a 5-day culture, FSH was obligatory for follicle growth and increased estradiol and testosterone secretion in a dose-dependent manner. DES (10(-6) M) caused the strongest decline in estradiol and testosterone levels but did not have detectable effects on either cAMP production or aromatase enzyme activity. GEN caused a prominent decrease in cAMP and testosterone levels without significant changes in secreted estradiol. The latter, apparently, was due to a dose dependent stimulation of aromatase enzyme activity in the presence of genistein. Both BP and OP decreased estradiol and testosterone secretion in a dose-dependent manner while no effect on aromatase activity was observed. OP, unlike BP, decreased forskolin-induced cAMP levels. Xenoestrogens at the used concentrations did not interfere with the growth and survival of the follicles. The results indicate that isolated ovarian follicles representing intact morphological and functional units offer a sensitive model system for elucidating the female specific reproductive effects of environmental chemicals. PMID- 15781296 TI - Absorption and evaporation of N,N-diethyl-m-toluamide from human skin in vitro. AB - The penetration of DEET through split-thickness cadaver skin was measured in non occluded Franz cells placed either in a fume hood or on a laboratory workbench. DEET, dissolved in a small volume of ethanol and spiked with (14)C radiolabel was applied to skin at doses from 0.02 to 11000 microg/cm(2). DEET penetration was greater for cells placed on the workbench, and the percentage of radioactivity penetrated after 72 h increased gradually with dose, for doses up to 680 microg/cm(2). At higher doses, it declined. Percent penetration ranged from 11.5 +/- 3.2% for a dose of 0.021 microg/cm(2) in the fume hood to 71.9 +/- 5.5% for a dose of 260 microg/cm(2) on the workbench. Results were interpreted in terms of a diffusion/evaporation model having three parameters-a solubility value for the chemical in the upper stratum corneum, M(sat); a mass transfer coefficient for evaporation, k(evap); and a characteristic time for diffusion, h(2)/D. The parameters obtained from fitting the model to the data (normalized to the fume hood environment) were M(sat) = 18 microg/cm(2) and k(evap) = 2.6 x 10(-5) cm/h. The value of h(2)/D decreased from 16 h at a DEET dose of 25 microg/cm(2) to 10 h at 1480 microg/cm(2), consistent with an increase in skin permeability of about 1.5-fold over this dose range. This effect was confirmed by means of an additional study in which skin samples pretreated with increasing amounts of unlabeled DEET were washed and redosed with (14)C-benzyl alcohol. A small (1.7 fold), but significant, increase in benzyl alcohol penetration with increasing amount of DEET was obtained. Thus, DEET enhanced its own skin permeation rate as well as that of another compound, but the effect was modest and not likely to be a major concern for compounds coadministered with DEET. PMID- 15781297 TI - Inhibition of renal Na+/H+ exchange in cadmium-intoxicated rats. AB - Chronic exposure to cadmium (Cd) results in bicarbonaturia, leading to metabolic acidosis. To elucidate the mechanism(s) by which renal bicarbonate reabsorption is inhibited, we investigated changes in renal transporters and enzymes associated with bicarbonate reabsorption in Cd-intoxicated rats. Cd intoxication was induced by subcutaneous injections of CdCl(2) (2 mg Cd/kg per day) for 3 weeks. Cd intoxication resulted in a significant reduction in V(max) of Na(+)/H(+) antiport with no changes in K(Na) in the renal cortical brush-border membrane vesicles (BBMV). Western blotting of BBM proteins and indirect immunohistochemistry in renal tissue sections, using an antibody against Na(+)/H(+) exchange-3 (NHE3), showed a diminished expression of NHE3 protein in the BBM. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that NHE3 mRNA expression was reduced in the renal cortex. The activity of carbonic anhydrase IV (CA IV) in BBM was not changed. The protein abundance of Na(+)-HCO(3)(-) cotransporter-1 (NBC1) in whole kidney membrane fractions was slightly attenuated, whereas that of the Na(+)-K(+)-ATPase alpha-subunit was markedly elevated in Cd-intoxicated animals. These results indicate that Cd intoxication impairs NHE3 expression in the proximal tubule, thereby reducing the capacity for bicarbonate reabsorption, leading to bicarbonaturia in an intact animal. PMID- 15781298 TI - Toxicity of sulphur mustard. PMID- 15781300 TI - Social cognition after head injury: sarcasm and theory of mind. AB - Closed head injury (CHI) is associated with communication difficulties in everyday social interactions. Previous work has reported impaired comprehension of sarcasm, using sarcastic remarks where the intended meaning is the opposite of the sincere or literal meaning. Participants with CHI in the present study were assessed using two types of sarcastic items, those with a directly opposite meaning and those with an indirect, non-literal but not directly opposite meaning. The CHI group was differentially poorer at comprehending sarcastic versus sincere remarks, although type of sarcastic materials did not influence performance. Errors involved not only literal interpretations, but also incorrect non-literal interpretations. Theory of mind (mentalising) was also assessed by comparing comprehension of human actions with control physical events. The CHI group was selectively impaired on the mentalising component of this task, and mentalising scores correlated with sarcasm comprehension. The implications of the findings for our understanding of impaired sarcastic comprehension after acquired brain injury are discussed. PMID- 15781302 TI - Conceptual information on objects' locations. AB - According to traditional views, basic and subordinate concepts elicit perceptual information, superordinate concepts abstract information. Two experiments showed that also superordinate concepts activate perceptual and contextual information. In Experiment 1 participants evaluated the adequacy of Scene- and Object-like locations ascribed to basic and superordinate concepts. Superordinate concepts were judged faster when paired with Scene-like locations, where many exemplars can coexist, than with Object-like locations. The results were replicated and extended in the second experiment with a location production task. Theoretical accounts for the results are discussed. PMID- 15781301 TI - Paralinguistic processing in children with callosal agenesis: emergence of neurolinguistic deficits. AB - Recent research revealed impaired processing of both nonliteral meaning and affective prosody in adults with agenesis of the corpus callosum (ACC) and normal intelligence. Since normal children have incomplete myelination of the corpus callosum, it was hypothesized that paralanguage deficits in children with ACC would be less apparent relative to their peers. The Familiar and Novel Language Comprehension Test (FANL-C) and Prosody Test were given to 18 children with ACC and normal intelligence and 17 controls matched for age (7-13 years), education, and IQ (83-122). When controlling for age, children with ACC were significantly poorer in comprehension of the precise meaning of both literal and nonliteral items on the FANL-C. Adults with ACC had previously been shown to have difficulty only on nonliteral items. The effect size for nonliteral comprehension in children with ACC was smaller than that seen in adults. There was only a trend for the child ACC group to perform worse on the recognition of affective prosody. Thus, while deficits in paralinguistic processing were apparent, children with ACC were not as clearly different from age peers as adults, and were equally deficient at comprehending literal and nonliteral expressions. The differences in results between adults and children with ACC are thought to reflect incomplete callosal development in normal children, and the importance of the corpus callosum in the early stages of the development of the ability to process literal language. PMID- 15781303 TI - Evidence of a highly specific relationship between rapid automatic naming of digits and text-reading speed. AB - This paper explores the specificity of the relationship between rapid automatic naming and reading fluency. Reading accuracy, rate, and fluency was measured among a sample of 67 children, the majority of whom were very poor readers. Regression analyses revealed that phonological processing tasks predicted reading accuracy and comprehension whereas rapid digit (but not picture) naming predicted reading accuracy and rate. After further controlling reading accuracy, digit naming was still a significant predictor of reading rate. This suggests that rapid alphanumeric naming is a highly specific predictor of reading rate and that rapid digit naming and phonological processing are distinct contributors to different aspects of reading in poor readers. PMID- 15781304 TI - Sequential processing in hemispheric word recognition: the impact of initial letter discriminability on the OUP naming effect. AB - The cerebral hemispheres have been proposed to engage different word recognition strategies: the left hemisphere implementing a parallel, and the right hemisphere, a sequential, analysis. To investigate this notion, we asked participants to name words with an early or late orthographic uniqueness point (OUP), presented horizontally to their left (LVF), right (RVF), or both fields of vision (BVF). Consistent with past foveal research, Experiment 1 produced a robust facilitatory effect of early OUP for RVF/BVF presentations, indicating the presence of sequential processes in lexical retrieval. The effect was absent for LVF trials, which we argue results from the disadvantaged position of initial letters of words presented in the LVF. To test this proposition, Experiment 2 assessed the discriminability of various letter positions in the visual fields using a bar-probe task. The obtained error functions highlighted the poor discriminability of initial letters in the LVF and latter letters in the RVF. To confirm that this asymmetry in initial letter acuity was responsible for the absent OUP effect for LVF presentations, Experiment 3 replicated Experiment 1 using vertical stimulus presentations. Results indicated a marked facilitatory effect of early OUP across visual fields, supporting our contention that the lack of OUP effect for LVF presentations in Experiment 1 resulted from poor discriminability of the initial letters. These findings confirm the presence of sequential processes in both left and right hemisphere word recognition, casting doubt on parallel models of word processing. PMID- 15781305 TI - The heterogeneity of picture-supported narratives in Alzheimer's disease. AB - This study describes discourse patterns produced by 46 Alzheimer disease (AD) patients and 53 normal elderly subjects in two picture-supported narratives. Nine measures derived from a cognitive model of discourse processing were obtained and submitted to cluster analysis. Results indicate that discourse patterns elicited from both stimuli were heterogeneous. Further, they fail to clearly differentiate between normal aging and AD in half of the AD patients. Discourse patterns are examined in light of various sources of heterogeneity including severity of cognitive impairment, subject characteristics, and task demands. The usefulness of discourse as a diagnostic and therapeutic tool is discussed. PMID- 15781306 TI - Neural correlates of lexicon and grammar: evidence from the production, reading, and judgment of inflection in aphasia. AB - Are the linguistic forms that are memorized in the mental lexicon and those that are specified by the rules of grammar subserved by distinct neurocognitive systems or by a single computational system with relatively broad anatomic distribution? On a dual-system view, the productive -ed-suffixation of English regular past tense forms (e.g., look-looked) depends upon the mental grammar, whereas irregular forms (e.g., dig-dug) are retrieved from lexical memory. On a single-mechanism view, the computation of both past tense types depends on associative memory. Neurological double dissociations between regulars and irregulars strengthen the dual-system view. The computation of real and novel, regular and irregular past tense forms was investigated in 20 aphasic subjects. Aphasics with non-fluent agrammatic speech and left frontal lesions were consistently more impaired at the production, reading, and judgment of regular than irregular past tenses. Aphasics with fluent speech and word-finding difficulties, and with left temporal/temporo-parietal lesions, showed the opposite pattern. These patterns held even when measures of frequency, phonological complexity, articulatory difficulty, and other factors were held constant. The data support the view that the memorized words of the mental lexicon are subserved by a brain system involving left temporal/temporo-parietal structures, whereas aspects of the mental grammar, in particular the computation of regular morphological forms, are subserved by a distinct system involving left frontal structures. PMID- 15781308 TI - Cognitive neuroscience and the English past tense: comments on the paper by Ullman et al. PMID- 15781311 TI - Tadpole collagenase, the single parent of such a large family. AB - This editorial review comments the development of the field of the matrix metalloproteinases that was initiated by the demonstration of the tadpole collagenase in 1962. PMID- 15781312 TI - Crystal structures of MMPs in complex with physiological and pharmacological inhibitors. AB - Matrix Metalloproteinases (MMPs) are a family of multidomain zinc endopeptidases that function in the extracellular space or attached to the cell membrane. Their proteolytic activity is controlled by the presence of endogenous inhibitors, the tissue inhibitors of matrix metalloproteinases (TIMPs), alpha-macroglobulin and others. Disruption of the proteinase-inhibitor balance is observed in serious diseases such as arthritis, tumor growth and metastasis, rendering the MMPs attractive targets for drug intervention by pharmacological inhibitors. The determination of MMP structures is of critical importance in order to understand their substrate preferences, dimerization events, and their association with matrix components and inhibitors. Thus, MMP structures may contribute significantly to the development of specific MMP inhibitors, which should allow precise control of individual members of the MMP family without affecting all members or the closely related metalloproteinases such as ADAMs and ADAMTSs. PMID- 15781313 TI - Assays of matrix metalloproteinases (MMPs) activities: a review. AB - Measurement of matrix metalloproteinase (MMP) activity often remains a challenge, mainly in complex media. Two sets of methods are currently used. The first one measures the hydrolysis of natural protein substrates (labeled or not) and includes the popular zymography. These techniques which are quite sensitive, cannot generally be carried out on a continuous basis. The second one takes mainly advantage of the increase of fluorescence, which is associated to the hydrolysis of initially quenched fluorogenic peptide substrates. Quite recently, another group, which is a compromise between the other two, has been developed. It measures the hydrolysis of synthetic triple-helical peptide substrates. These different methods are described and discussed. PMID- 15781314 TI - Collagenases in cancer. AB - Three mammalian collagenases (MMP-1, MMP-8, and MMP-13) belong to family of matrix metalloproteinases and are the principal secreted endopeptidases capable of cleaving collagenous extracellular matrix. In addition to fibrillar collagens, collagenases can cleave several other matrix and non-matrix proteins including growth factors, and this way regulate cell growth and survival. Collagenases are important proteolytic tools for extracellular matrix remodeling during organ development and tissue regeneration, but they also apparently play important roles in many pathological situations and tumor progression and metastasis. Because of their potentially destructive characteristics the expression and activity of collagenases are strictly controlled. Synthesis of collagenases is regulated by extracellular signals via cellular signal transduction pathways at transcriptional and post-transcriptional level. Collagenases are synthesized as inactive pro-forms, and once activated, their activity is inhibited by specific tissue inhibitors of metalloproteinases, TIMPs, as well as by non-specific proteinase inhibitors. In this review we discuss the current view on the role of collagenases in tumor growth, invasion, and metastasis, as a basis for their feasibility in diagnosis and prognostication, as well as therapeutic targets in cancer patients. PMID- 15781315 TI - Gelatinases (MMP-2 and -9) and their natural inhibitors as prognostic indicators in solid cancers. AB - Neoplastic growth and dissemination involve increased proteolytic activity that is able to escape the regulative elements. Matrix metalloproteinases (MMPs), particularly gelatinases A and B (MMP-2 and -9), play a role in tumor invasion and angiogenesis, and they participate in cancer progression in several neoplasias. The expression of tissue inhibitors of gelatinases, TIMPs-1 and -2, has also been shown to be associated with the clinical course in some cancers. The prognostic value of these markers, however, seems to vary a great deal in different neoplastic diseases. In this review, the impact of the gelatinases and their inhibitors on the clinical course in several solid cancers is evaluated based on the growing data from recent clinical studies. The clinical data most often explore the overexpression of mRNA or immunoreactive protein in tumor tissue, or measure the concentration of the circulating proteinase or its inhibitor in pretreatment or follow-up serum samples. The growing amount of recent clinical data suggests that the impact of gelatinases on treatment decisions should be tested in clinical trials. PMID- 15781316 TI - From a unique cell to metastasis is a long way to go: clues to stromelysin-3 participation. AB - Stromelysin-3 (ST3) overexpression is associated with poor patient clinical outcome in numerous carcinomas. The ST3 is expressed by peritumoral fibroblast like cells. Review of the literature shows that ST3 is an active partner of cancer cells along the whole natural cancer history, and is essential for optimal tumor development as it reduces death of cancer cells invading adjacent connective tissues at the primary tumor site. Paradoxically, ST3 lowers metastasis development in vivo in mice. However, this beneficial effect does not counterbalance the deleterious anti-apoptotic function of ST3. PMID- 15781317 TI - Role of matrix metalloproteinases in melanoma cell invasion. AB - Cutaneous melanomas are notorious for their tendency to metastasize. Essential steps in this process are the degradation of basement membranes and remodeling of the extracellular matrix (ECM) by proteolytic enzymes such as matrix metalloproteinases (MMPs), which are regulated by their tissue inhibitors (TIMPs). An MMP expression is not restricted to tumor cells but is also found in stromal cells, indicating that stroma-derived proteases may contribute to melanoma progression. The MMPs have been shown to interact with a broad range of non-matrix proteins including adhesion molecules, growth factors and mediators of angiogenesis and apoptosis. In this review, we evaluate new insights into the interplay of MMPs and their molecular partners in melanoma progression. PMID- 15781318 TI - Functional imaging of pericellular proteolysis in cancer cell invasion. AB - Proteolytic interactions between cells and extracellular matrix (ECM) are involved in many physiological and pathological processes, such as embryogenesis, wound healing, immune response, and cancer. The visualization of cell-mediated proteolysis towards ECM is thus required to understand basic mechanisms of tissue formation and repair, such as the breakdown and structural remodelling of ECM, inflammatory changes of tissue integrity, and the formation of proteolytic trails by moving cells. A panel of synergistic techniques for the visualization of pericellular proteolysis in live and fixed samples allow monitoring the of proteolytic tumor cell invasion in three-dimensional (3D) fibrillar collagen matrices in vitro. These include the quantification of collagenolysis by measuring the release of collagen fragments, the detection of protease expression and local activity by dequenching of fluorogenic substrate, and the staining of cleavage-associated neoepitopes together with changes in matrix structure. In combination, these approaches allow the high-resolution mapping of pericellular proteolysis towards ECM substrata including individual focal cleavage sites and the interplay between cell dynamics and alterations in the tissue architecture. PMID- 15781319 TI - Tumor-stroma interactions: their role in the control of tumor cell invasion. AB - The development and progression of tumors result from the concerted activity not only of tumor cells with neighboring cells e.g., fibroblasts and inflammatory cells. Host-tumor interactions are considered critical in tumor invasion and metastasis. In vitro studies as well as established in vivo models have analysed the reciprocal effects of tumor-host interactions for the tumor invasion process. These studies have shown that modifications in the extracellular matrix composition surrounding the tumors as well as alterations in the expression of tumor cell receptors or in the expression of growth factors/cytokines and proteases, are critical regulators of a developing tumor. We shortly review the most important and well characterized mechanisms involved in the progression of tumor cells through tissues, especially those participating in cellular communication, cell adhesion, and proteolysis. PMID- 15781320 TI - Membrane type-matrix metalloproteinases and tumor progression. AB - Matrix metalloproteinases (MMPs) are a family of zinc endopeptidases that process growth factors, growth factor binding proteins, cell surface proteins, degrade extracellular matrix (ECM) components and thereby play a central role in tissue remodeling and tumor progression. Membrane-type matrix metalloproteinases (MT MMPs) are a recently discovered subgroup of intrinsic plasma membrane proteins. Their functions have been extended from pericellular proteolysis and control of cell migration to cell signaling, control of cell proliferation and regulation of multiple stages of tumor progression including growth and angiogenesis. This review sheds light on the new functions of MT-MMPs and their inhibitors in tumor development and angiogenesis, and presents recent investigations that document their influence on various cell functions. PMID- 15781321 TI - Effects of basic fibroblast factor (bFGF) on MMP-2, TIMP-2, and type-I collagen levels in human lung carcinoma fibroblasts. AB - Matrix metalloproteinases (MMP's) and tissue inhibitors of metalloproteinases (TIMP's) possess a preponderant role in the metabolism of the major extracellular matrix protein, collagen, and are thought to be important in the mechanism of tumor invasion. Lung cancer occupies the first position in mortality and the second position in incidence, among all cancers. In the present investigation, we studied the effect of basic fibroblast growth factor (bFGF) on collagen, matrix metalloproteinase-2 (MMP-2), and tissue metalloproteinase inhibitor-2 (TIMP-2) levels in normal and carcinoma lung tissue fibroblast cultures. MMP-2 was selected because of its high specificity in the degradation of type IV collagen, major component of the basal membrane. The effect of bFGF on MMP-2, TIMP-2, total collagen, and type I collagen levels of normal and carcinoma lung fibroblast cultures was investigated at 0, 10, and 100 ng/ml. Statistical analysis was carried out using the Mann-Whitney-U test and possible correlations were searched using the Spearman correlation analysis method. MMP-2, TIMP-2, total collagen, and type-1 collagen levels based on cell counts (10(3) cells) showed no statistically significant differences between the carcinoma and normal fibroblast cultures. However, positive correlations were found between MMP-2 and TIMP-2 in normal (P = 0.016) and carcinoma (P = 0.001) tissue fibroblast cultures. Positive correlations were also found between total collagen and TIMP-2 levels in normal and carcinoma tissue fibroblast cultures (P = 0.002 and P = 0.032). Total collagen and TIMP-2 levels also showed positive and strong correlations in all cultures except in 100 ng/ml bFGF concentrations. In addition, type I collagen and MMP-2 levels showed positive significant correlations only in normal and carcinoma control cultures, while type I collagen and TIMP-2 levels showed positive correlations in all cultures except carcinoma fibroblasts at 100 ng/ml bFGF. It may be concluded that bFGF does not affect MMP-2, TIMP-2, total collagen, and type-1 collagen levels in fibroblast cultures grown from human carcinoma and normal lung tissues. However, bFGF was noted, in vitro, to disturb the equilibrium which normally exists between the four parameters, both in normal and carcinoma tissue fibroblasts. PMID- 15781322 TI - Matrikines in the regulation of extracellular matrix degradation. AB - The term "matrikines" was coined for designating peptides liberated by partial proteolysis of extracellular matrix macromolecules, which are able to regulate cell activities. Among these peptides, some of them may modulate proliferation, migration, protease production, or apoptosis. In this review, we summarize the activity of matrikines derived from elastin and interstitial or basement membrane collagens on the regulation of matrix metalloproteinases expression and/or activation, and on the plasminogen/plasmin system. Due to their activity, matrikines may play a significant role in physiological or pathological processes such as wound healing or tumor invasion. PMID- 15781323 TI - EMMPRIN/CD147, an MMP modulator in cancer, development and tissue repair. AB - Matrix metalloproteinases (MMPs) play a central role in normal tissue remodeling and disease, they regulate tumor microenvironment and their expression is increased in most human cancers. Targeting their activity remains a major challenge. Their production and activation is tightly regulated by complex mechanisms that include cytokines and growth factors, cell-matrix and cell-cell interactions. The observations of increased MMP level at the epithelio-stromal interface led to the identification of EMMPRIN/CD147, a membrane spanning molecule highly expressed in tumor cells, that stimulates MMPs production in neighboring fibroblasts. Later studies have shown that EMMPRIN can also induce MMP in the same population of cells. Elevated EMMPRIN level was detected in numerous malignant tumors and has been correlated with tumor progression in experimental and clinical conditions. The presence and modulation of EMMPRIN in normal tissues associated with increased MMP expression suggests that this EMMPRIN-mediated MMP induction could be a common mechanism in non-tumoral physiological and/or pathological situations. Targeting EMMPRIN in cancer and other pathological conditions such arthritis and ulceration appears a promising future therapeutic strategy, but requires a better understanding of its mode of action and regulation. Potential regulators that influence EMMPRIN level and its MMP inducing activity include growth factors, hormones, glycosylation and membrane shedding. This review will discuss the recent findings concerning these diverse regulatory mechanisms in various physiological and pathological situations. PMID- 15781324 TI - Regulation of matrix metalloproteinase (MMP) activity by the low-density lipoprotein receptor-related protein (LRP). A new function for an "old friend". AB - Matrix metalloproteinases (MMPs) are essential contributors to a microenvironment that promotes tumour progression. During the two last decades, inhibition of MMPs has become the focus of considerable interest for cancer therapy, and numerous synthetic metalloproteinase inhibitors have been developed by the pharmaceutical industry. However, clinical trials have shown disappointing efficacy or unexpected toxicity and new targets are thus eagerly awaited. The identification of endocytic clearance of several MMPs by the low-density lipoprotein receptor related protein (LRP) might provide insight into novel strategies for controlling MMP level during malignant processes. This review attempts to summarize recent aspects on the cellular and molecular basis of LRP-mediated endocytic disposal of MMPs. PMID- 15781325 TI - Beneficial and detrimental influences of tissue inhibitor of metalloproteinase-1 (TIMP-1) in tumor progression. AB - Tissue inhibitor of metalloproteinase-1 (TIMP-1) is one representative of the natural matrix metalloproteinase (MMP) inhibitor family, encompassing four members. It inhibits all MMPs, except several MT-MMPs, and a disintegrin with a metalloproteinase domain (ADAM)-10 with Kis < nM. Unexpectedly, its upregulation was associated to poor clinical outcome for several cancer varieties. Such finding might be related to the growth-promoting and survival activities of TIMP 1 for normal and cancer cells. In most cases, such properties are MMP-independent and binding of TIMP-1 to an unknown receptor system can trigger JAK (or FAK)/PI3 kinase/Akt/bad-bclX2 (erythroid, myeloid, epithelial cell lines) or Ras/Raf1/FAK (osteosarcoma cell line) signaling pathways. The relationship between viral infection and TIMP-1 expression is here underlined. Thus, TIMP-1 might display a dual influence on tumor progression; either beneficial by inhibiting MMPs as MMP 9 and by impairing angiogenesis or detrimental by favoring cancer cells growth or survival. We consider that the proMMP-9/TIMP-1 balance is of critical importance in early events of tumor progression, and might show promise as diagnostic and prognostic marker of malignancy. PMID- 15781326 TI - Hydroxamate-based peptide inhibitors of matrix metalloprotease 2. AB - There is major interest in designing inhibitors for matrix metalloproteinase 2 (MMP-2, gelatinase A) since this enzyme is known to be involved in pathological processes such as tumor invasion or rheumatoid arthritis. The majority of MMP-2 inhibitor candidate drugs block the active site of MMP-2 by binding to its catalytic Zn2+ ion through a chelating (hydroxamate, sulphonate etc.) group. Despite the general interest in designing MMP-2 inhibitors, the results with many of the drug candidates were disappointing, their failure was usually explained by cross-reactions with other MMPs. One way to enhance MMP-2 selectivity is to design inhibitors that interact with both the active site and exosites such as the fibronectin type II (FN2) domains of the enzyme. In the present work, we have examined the inhibitory potential and MMP-2 selectivity of hydroxamates of three groups of peptides known to bind to the collagen-binding FN2 domains of MMP-2. The first type of peptides consisted of collagen-like (Pro-Pro-Gly)(n) repeats, peptides of the second group were identified from a random 15-mer phage display library based on their binding to immobilized FN2 domains of MMP-2. A hydroxamate of peptide p33-42, known to bind to the third FN2 domain of MMP-2 has also been tested. Our studies have shown that these compounds inhibited MMP-2 with IC50 values of 10-100 microM. The fact that their inhibitory potential was nearly identical for MMP-2del, a recombinant version of MMP-2 that lacks the FN2 domains, suggests that inhibition is not mediated by their binding to FN2 domains. It seems likely that the failure to exploit interaction with the FN2 domains is due to the fact that the FN2 domains and the catalytic domain of MMP-2 tumble independently, therefore only a tiny fraction of the conformational isomers can bind peptide hydroxamates via both the active site and the FN2 domain(s). PMID- 15781327 TI - Future challenges facing the development of specific active-site-directed synthetic inhibitors of MMPs. AB - Despite a deep knowledge on the 3D-structure of several catalytic domains of MMPs, the development of highly specific synthetic active-site-directed inhibitors of MMPs, able to differentiate the different members of this protease family, remains a strong challenge. Due to the flexible nature of MMP active site, the development of specific MMP inhibitors will need to combine sophisticated theoretical and experimental approaches to decipher in each MMP the specific structural and dynamic features that can be exploited to obtain the desired selectivity. PMID- 15781328 TI - Stem cell biology and the plasticity polemic. AB - Characterization of a cord blood derived unrestricted somatic stem cell (USSC) with capacity to differentiate into hematopoietic and nonhematopoietic tissues in the absence of cell fusion has highlighted the great potential of stem cell plasticity. A great variety of stem cell types have been defined and even the most pure marrow stem cells are highly heterogeneous. Data suggest that stem cells may exist in a continuum with continually and reversibly changing phenotype. These cells also possess a capacity to produce lung, liver, skin, and skeletal muscle under conditions of tissue injury. Arguments raised against the significance of adult marrow to nonmarrow conversions including the importance of cell fusion appear fallacious. We are at the beginning of an exciting and burgeoning field of research with great clinical potential. PMID- 15781330 TI - GPIIb (CD41) integrin is expressed on mast cells and influences their adhesion properties. AB - OBJECTIVES: GPIIb integrin expression has been found on platelets and megakaryocytes, and more recently on immature hematopoietic progenitors. We set out to investigate expression of GPIIb in other hematopoietic cell lineages and, having detected it on mast cells, aimed to determine what possible role it might perform. METHODS: We have made use of cultured human and murine bone marrow mast cells (BMMC) in order to characterize the expression of GPIIb. Further, BMMC cultures from wild type and GPIIb deficient (gpIIb-/-) mice were used for comparison of the adhesive properties mediated by this receptor. Finally, peritoneal mast cells were analyzed from both wild type and (gpIIb-/-) mice. RESULTS: We demonstrate expression of GPIIb on cultured BMMC. Using cells derived from mice homozygous for a null allele of gbIIb we show that the absence of GPIIb has no effect on mast cells with respect to a number of measures of cell growth and differentiation. However, loss of GPIIb on BMMC results in an increase in surface expression of aV integrin, the alternative partner of GPIIIa. CONCLUSION: The results in this study demonstrate that GPIIb is expressed in human and murine mast cells. A function for GPIIb on mast cells is suggested by the altered adhesion of gbIIb-/- BMMC to fibronectin- and vitronectin-coated surfaces. Moreover, comparison of mast cells from the peritoneal cavity of wild type and gbIIb-/- mice indicates that GPIIb could influence the in vivo differentiation or homing of tissue mast cells. PMID- 15781329 TI - Visualizing PU.1 activity during hematopoiesis. AB - OBJECTIVE: PU.1 is a critical transcription factor for hematopoietic development that is required for the early differentiation of myeloid, erythroid, and B lineage cells. To gain a better insight into PU.1 function, we performed a comprehensive analysis of PU.1 gene activity in the hematopoietic system, using a green fluorescent protein reporter mouse line. METHODS: We used flow cytometry to analyze green fluorescent protein (GFP) expression, along with various cell surface markers, in heterozygote mice that harbor a GFP reporter knocked into exon1 of the PU.1 gene. Phenotypic and functional properties of GFP+ and GFP- precursors were studied. RESULTS: We show that PU.1 is dynamically and heterogeneously expressed in many hematopoietic lineages, from the stem cell stage to terminally differentiated cells, suggesting that PU.1 is not only important in early differentiation events but also may play a role in mature hematopoietic cell function. Further, examination of GFP+ vs GFP- populations shows that differentiation, but not commitment, to the myeloid lineage requires PU.1. In contrast, B cell commitment is associated with low levels of PU.1 expression. CONCLUSION: Our study provides a detailed visualization of PU.1 gene activity in hematopoietic cells, and shows that highly dynamic regulation of PU.1 accompanies cell fate decisions during hematopoiesis. PMID- 15781331 TI - Thrombopoietin alone or in the presence of stem cell factor supports the growth of KIT(CD117)low/ MPL(CD110)+ human mast cells from hematopoietic progenitor cells. AB - OBJECTIVES: Thrombopoietin (TPO) is known to promote platelet number, have growth promoting potential for human megakaryocytes (HuMKs), and increase erythrocyte, monocyte, mast cell, and granulocyte numbers in the presence of additional growth factors. We explored the ability of TPO alone or in the presence of stem cell factor (SCF) to support human mast cells (HuMCs). METHODS: CD34+ pluripotent and CD34+/CD117+/CD13+ HuMC progenitor cells were cultured in rhTPO and examined for HuMCs. Similarly, we added rhTPO to CD34(+) cells cultured in stem cell factor (SCF), which promotes HuMC development. RESULTS: When CD34+ cells were cultured in 10 ng/mL rhTPO and 10 ng/mL rhSCF, TPO enhanced HuMC numbers compared to rhSCF alone. Higher concentrations of rhTPO (50 ng/mL) in the presence of 100 ng/mL rhSCF inhibited the rhSCF-dependent subpopulation of CD117high HuMCs, while promoting CD117low HuMCs. Human CD34+/CD117+/CD13+ cells cultured in rhTPO alone for 1 to 2 weeks differentiated into CD41+/CD110+ HuMKs (85-90%) and FcepsilonRI+/CD117low/CD13+ HuMCs (5-10%). RhTPO-induced HuMCs expressed the TPO (CD110) receptor, tryptase, and chymase and survived when recultured in rhSCF. CONCLUSION: The effect of TPO on HuMCs in the presence of rhSCF varies, depending on the relative concentration of each growth factor, while TPO alone or in combination with rhSCF supports a unique population of CD117low/CD110+ HuMCs. PMID- 15781332 TI - All-trans retinoic acid (ATRA) enhances maintenance of primitive human hematopoietic progenitors and skews them towards myeloid differentiation in a stroma-noncontact culture system. AB - OBJECTIVE: We have previously shown that hematopoietic progenitor cells (HPCs) from umbilical cord blood (UCB) can be maintained in a cytokine-supplemented stroma-noncontact (SNC) system. Here, we tested if all-trans retinoic acid (ATRA), known to improve expansion of murine hematopoietic stem cells, would enhance human HPC maintenance in a SNC culture system. METHODS: CD34+CD38-Lin- cells from UCB were cultured in transwells above AFT024 in the presence of Flt-3 ligand (FLT) and thrombopoietin (TPO), with or without ATRA. Total nucleated cells (TNC), colony-forming units (CFUs), long-term culture-initiating cells (LTC ICs), myeloid-lymphoid initiating cells (ML-ICs) and SCID repopulating cells (SRCs) were evaluated 1 to 5 weeks after culture. RESULTS: All-trans retinoic acid (1 mumol/L) reduced expansion of CD34+CD38-Lin- TNC and CFUs after 2 to 5 weeks of culture. However, it significantly increased LTC-IC expansion after 1 to 3 and, even more so, 5 weeks of culture. ATRA also increased recovery of more primitive ML-ICs and SRCs. Increased HPC recovery appeared dependent on the presence of stromal cells, as LTC-IC expansion was significantly reduced when ATRA was added to stroma-free cultures. CONCLUSION: All-trans retinoic acid increases expansion of early HPCs in a stromal cell-dependent fashion. PMID- 15781333 TI - Soluble factor(s) from bone marrow cells can rescue lethally irradiated mice by protecting endogenous hematopoietic stem cells. AB - OBJECTIVE: Ionizing radiation-induced myeloablation can be rescued via bone marrow transplantation (BMT) or administration of cytokines if given within 2 hours after radiation exposure. There is no evidence for the existence of soluble factors that can rescue an animal after a lethal dose of radiation when administered several hours postradiation. We established a system that could test the possibility for the existence of soluble factors that could be used more than 2 hours postirradiation to rescue animals. MATERIALS AND METHODS: Animals with an implanted TheraCyte immunoisolation device (TID) received lethal-dose radiation and then normal bone marrow Lin- cells were loaded into the device (thereby preventing direct interaction between donor and recipient cells). Animal survival was evaluated and stem cell activity was tested with secondary bone marrow transplantation and flow cytometry analysis. Donor cell gene expression of five antiapoptotic cytokines was examined. RESULTS: Bone marrow Lin- cells rescued lethally irradiated animals via soluble factor(s). Bone marrow cells from the rescued animals can rescue and repopulate secondary lethally irradiated animals. Within the first 6 hours post-lethal-dose radiation, there is no significant change of gene expression of the known radioprotective factors TPO, SCF, IL-3, Flt-3 ligand, and SDF-1. CONCLUSION: Hematopoietic stem cells can be protected in lethally irradiated animals by soluble factors produced by bone marrow Lin- cells. PMID- 15781334 TI - Programmed cell death is an intrinsic feature of MDS progenitors, predominantly found in the cluster-forming cells. AB - OBJECTIVE: Bone marrows (BM) of myelodysplastic syndrome (MDS) patients show increased proliferation and premature programmed cell death (PCD) in vivo as well as in vitro. We explored the proliferative capacity and apoptotic propensity of CD34+ progenitor cells of MDS patients excluding accessory cell interference. MATERIALS AND METHODS: CD34+/CD3-/CD19- cells of 5 MDS patients and 5 normal BM were sorted as single cells into single wells and were cultured in liquid medium. Wells were evaluated on days 4, 7, 10, and 14. PCD was determined by staining with annexin V-FITC. Growth rate and cell doubling time (Td) were calculated for each colony-forming cell. RESULTS: Normal BM CD34+ cells formed clusters and colonies and both showed increasing PCD in time, although within colonies the degree of apoptosis was twice as high (about 25%) as compared with clusters at all time points. In MDS increased cluster formation was observed at all evaluation points when compared to normal BM, whereas the number of colonies was markedly reduced (1/7 of normal). These colonies were also smaller, usually smaller than 100 cells. Significantly enhanced levels of PCD of clusters (53-79%) in combination with longer cell doubling times explain this slower formation of smaller colonies. Surprisingly, these colonies showed considerably lower levels of PCD (7-32%) as compared to normal (1-48%, median values). CONCLUSIONS: In the absence of stromal influences and accessory cells, this study in MDS patients showed intrinsically enhanced proliferation and apoptosis of cluster-forming cells, as the opposite was true for colony-forming cells. PMID- 15781335 TI - WASP- mice exhibit defective immune responses to influenza A virus, Streptococcus pneumoniae, and Mycobacterium bovis BCG. AB - OBJECTIVE: To quantify the immune response of WASP- mice to three different pathogens: influenza A virus, Streptococcus pneumoniae, and Mycobacterium bovis. METHODS: Primary and secondary T-cell responses to influenza A virus were quantified via tetramer assays. Viral clearance from lung was also measured. Lethality of intranasal inoculation with luminescent S. pneumoniae was assessed by dose escalation and direct luminescence imaging. After intravenous inoculation with M. bovis, residual mycobacteria in lung, liver, and spleen were measured by standard culture methods. RESULTS: The reduced secondary T-cell response to influenza A virus correlates with a relative but not absolute loss of splenic T and B cells similar to that seen in clinical Wiskott-Aldrich Syndrome (WAS), and slower clearance of virus from lung. The reduced magnitude of the secondary T cell response correlates with a progressive loss of influenza-specific T cells after primary inoculation. WASP- mice show an increased susceptibility to lethal pneumonia after intranasal inoculation with S. pneumoniae, which is among the most frequent causes of clinical complications in WAS patients. WASP- mice clear M. bovis bacille Calmette-Guerin (BCG) more slowly from lung, liver, and spleen. Bone marrow-derived macrophages, however, show normal ex vivo cytokine secretion in response to M. bovis. CONCLUSIONS: These results demonstrate that WASP- mice are functionally immunodeficient in regard to three different pathogens, and provide relevant end points for the study of treatment modalities in this model. They also suggest a specific physiologic mechanism, failure to accumulate memory T cells, for at least one of the defective immune responses. PMID- 15781336 TI - T cells armed with anti-CD3 x anti-CD20 bispecific antibody enhance killing of CD20+ malignant B cells and bypass complement-mediated rituximab resistance in vitro. AB - OBJECTIVE: Resistance to rituximab, a chimeric monoclonal antibody that binds to CD20, is a major limitation for the successful treatment of patients with non Hodgkin lymphoma and other CD20+ B-cell malignancies. To circumvent rituximab resistance in these patient populations, we have constructed a bispecific antibody (BiAb), anti-CD3 x anti-CD20 (CD20Bi), that combines rituximab targeting with non-major histocompatibility complex (non-MHC)-restricted cytotoxicity mediated by activated T cells (ATC). MATERIALS AND METHODS: Activated T cells were obtained from anti-CD3 activated peripheral blood mononuclear cells (PBMC) of normal donors or the leukapheresis products of patients by culturing in the presence of interleukin-2 for 6-14 days. After ATC expansion, the cells were armed with CD20Bi. Killing activity was evaluated by 51Cr-release assay. RESULTS: Arming ATC with as little as 5 ng CD20Bi/10(6) cells significantly increased cytotoxicity above unarmed ATC. CD20Bi-armed ATC (50 ng/10(6) cells) efficiently lysed CD20+ cell lines at E:T of 6.25-50, but not the nonhematologic, CD20- SK-BR 3 cell line. High levels of cytotoxicity mediated by CD20Bi-armed ATC (p < 0.05) could not be blocked by an 8000-fold excess of soluble rituximab. CD20Bi-armed ATC in the presence of complement killed ARH-77 cells, a rituximab-complement pathway-resistant multiple myeloma, significantly (p < 0.05) better than rituximab or unarmed ATC, suggesting that CD20Bi-armed ATC may be clinically effective for treatment of rituximab-resistant CD20+ hematologic malignancies. CONCLUSIONS: Our findings demonstrate that CD20Bi-armed ATC enhance cytotoxicity against CD20+ B-cell lines and circumvent complement-mediated rituximab resistance, providing a strong rationale for this immune-based strategy for the treatment of rituximab-refractory CD20+ B-cell malignancies. PMID- 15781337 TI - Enhanced function with decreased internalization of carboxy-terminus truncated CXCR4 responsible for WHIM syndrome. AB - OBJECTIVE: WHIM (warts, hypogammaglobulinemia, recurrent bacterial infection, myelokathexis) syndrome is an autosomal dominant immune deficiency with severe chronic neutropenia and marrow neutrophil apoptosis. Carboxy-termini truncating mutations in the chemokine receptor CXCR4 have been identified in WHIM patients. We created a retrovirus encoding mutated CXCR4 (truncating point mutation 1000C- >T [R334X] inherited heterozygously in several WHIM patients) in order to transducer healthy human CD34 stem cells and K562 to overexpress mutated CXCR4 and determined its effect on receptor responses to stromal-derived factor-1 (SDF1). METHODS: Retrovirus vector was engineered to coexpress WHIM-associated R334X mutated CXCR4 together with green fluorescent protein (GFP). Control vectors included similar constructs with wild-type CXCR4 (WT-CXCR4) or only GFP. CD34+ cells and K562 were transduced with these vectors. Populations of 100% transduced K562 were established by sorting GFP+ cells by flow cytometry. We performed migration and calcium flux assays of transduced CD34+ cells and transduced/sorted K562. We also examined receptor recycling in response to SDF1. RESULTS: Healthy human CD34+ cells and/or human erythroleukemia K562 cells transduced to express mutated CXCR4, WT-CXCR4, or GFP alone demonstrated that mutated CXCR4 was associated with enhanced calcium flux and enhanced migration. There was also decreased receptor internalization and enhanced recovery of surface mutated CXCR4 in response to SDF1 compared with WT-CXCR4. CONCLUSION: We propose that decreased internalization of WHIM-associated mutated CXCR4 leads to prolongation/enhancement of signaling in response to SDF1 and that this may provide the biochemical basis for the autosomal dominant abnormalities of cell trafficking and function associated with WHIM syndrome. PMID- 15781338 TI - Src family tyrosine kinases are activated by Flt3 and are involved in the proliferative effects of leukemia-associated Flt3 mutations. AB - OBJECTIVE: The hematopoietic growth factor receptor, Fms-like tyrosine kinase-3 (Flt3), modulates survival and proliferation of myeloid and B-cell precursors. Activating mutations of Flt3 are the most common molecular abnormalities in acute myeloid leukemia (AML) and have an apparent role in leukemogenesis. However, signaling pathways mediating Flt3 effects are incompletely understood. The role of Src kinases is unknown, although some, such as Lyn, have also been linked to leukemogenesis. This study examines the role of Src kinases in Flt3 signaling and the oncogenic effects of leukemia-associated Flt3 mutations. MATERIALS AND METHODS: We examined the activation and functional roles of Src kinases in human leukemic myeloid cell lines expressing wild-type Flt3 or a constitutively active mutant, and in cells stably transduced with human wild-type or mutant Flt3. RESULTS: Flt3 ligand stimulation of wild-type Flt3 increased phosphorylation of Src kinase Lyn. Constitutive Lyn phosphorylation and activation was found in cells expressing constitutively active Flt3 mutants. Src kinases are implicated in downregulation of closely related receptors, but Src inhibitors had no effect on ligand-stimulated Flt3 degradation, or on the rapid degradation of an Flt3 mutant. However, growth-factor-independent proliferation resulting from mutant Flt3 expression did depend on the activity of Src kinases. CONCLUSION: Our studies reveal for the first time the involvement of Src kinases in Flt3 signaling, with activation of Lyn by constitutively active Flt3 mutants as well as ligand-stimulated wild-type receptor, and show that Src kinase inhibitors block proliferative effects of Flt3 mutants found in AML. Thus, Src kinases may represent targets for inhibitor therapy in Flt3-related AML. PMID- 15781340 TI - HIV-associated lymphoma successfully treated with peripheral blood stem cell transplantation. AB - OBJECTIVE: To evaluate feasibility, safety, and efficacy of peripheral blood stem cell collection (PBSCC) and autologous stem cell transplantation (ASCT), to treat patients diagnosed of high-risk or relapsed HIV-associated lymphoma (HIV+ Ly), responding to highly active antiretroviral therapy (HAART). METHODS: Prospective and multicentric study in patients with high-risk or relapsed chemosensitive HIV+ Ly, candidate for consolidation with ASCT. Eligibility criteria were similar to those of HIV- lymphoma. HAART was aimed to be maintained during the procedure. RESULTS: Fourteen patients were admitted. Adequate PBSCC was obtained from all patients (median CD34+ cells was 4.7 x 10(6)/kg). Three patients died before ASCT; two had disease progression and one died from VHC-liver failure. Eleven transplanted patients showed neutrophil engraftment after a median time of 16 days (range, 9-33 days), and nine patients showed platelet engraftment after a median time of 20 days (range, 11-36 days). CD4+ cell counts and HIV viral load (VL) were appropriately preserved along the procedure. No patients died from treatment-related complications. One patient died from lymphoma progression (day +19), and another died in complete remission (CR) with undetectable VL, 15 months after transplant, due to infection. One patient relapsed at 32 months after ASCT. The remaining eight patients are alive in CR with an event-free survival of 65% and a median follow-up of 30 months after ASCT (range, 7-36 months). CONCLUSIONS: These results show that feasibility, safety, and efficacy of PBSCC and ASCT in HIV+ Ly patients responding to HAART are similar to those observed in the HIV- lymphoma setting. PMID- 15781339 TI - B lymphocyte reconstitution after hematopoietic stem cell transplantation: functional immaturity and slow recovery of memory CD27+ B cells. AB - OBJECTIVE: Functional recovery of B lymphocytes after hematopoietic stem cell transplantation (HSCT) can take up to 2 years. HSCT recipients may obtain protective titers of pathogen-specific antibody through vaccination, but optimal timing of reimmunization remains to be defined. PATIENTS AND METHODS: In this study, we evaluated the reconstitution of B-cell number and activity in 139 children given HSCT, by B-cell subset phenotyping and in vitro immunoglobulin (Ig) production. RESULTS: Patients were longitudinally studied at 3, 6, 12, and 18 to 24 months after transplantation. At all time points, recipients displayed a significantly higher percentage of naive (IgD+CD27-) B cells and showed significantly lower production of stimulated in vitro Ig as compared to healthy controls. Moreover, during follow-up, we observed an increase in the proportion of patients who had CD27+ B subsets and who were able to mount in vitro Ig production greater than the 5th percentile. CONCLUSION: Similar to what has been described in adults, most children lack memory B cells and produce low amounts of Ig. However, the number of B cells, as well as their function, gradually recovered over time and the spread of data we observed suggests that the reimmunization schedule should be individualized for each patient. It remains to be defined in a prospective clinical study the time point at which a patient should start reimmunization. A reasonable hypothesis to be explored is the time point at which a percentage of memory B cells greater than the 5th percentile of normal controls is reached. PMID- 15781341 TI - Differential effects of a novel IFN-zeta/limitin and IFN-alpha on signals for Daxx induction and Crk phosphorylation that couple with growth control of megakaryocytes. AB - OBJECTIVE: Although a novel IFN-zeta/limitin uses IFN-alpha/beta receptor, it lacks some common activities of type I IFNs. We compared effects on megakaryocyte proliferation and differentiation as well as signals for their biological activities. MATERIALS AND METHODS: Recombinant IFN-zeta/limitin and IFN-alpha titrated with a cytopathic effect dye binding assay, were used in this study. Colony assays and serum-free suspension cultures for megakaryocytes were performed to compare their growth inhibitory effects. To analyze signals, megakaryocytes cultured in serum-free suspension cultures were stimulated and Western blotted with the indicated antibody. RESULTS: Both IFN-zeta/limitin and IFN-alpha suppressed the proliferation of megakaryocyte progenitors without influencing their differentiation. However, much higher concentrations of IFN zeta/limitin were required for the growth inhibition than IFN-alpha. The growth inhibition by IFN-zeta/limitin and IFN-alpha was significantly reduced when either Tyk2 or STAT1 was disrupted. In addition, the antisense oligonucleotides against Crk and Daxx, downstream molecules of Tyk2, greatly rescued the IFN zeta/limitin- and IFN-alpha-induced reduction of megakaryocyte colony numbers. In cultured megakaryocytes, IFN-zeta/limitin induced the expression of SOCS-1 as strongly as IFN-alpha. However, IFN-zeta/limitin induced weaker phosphorylation of Crk and lower induction of Daxx than IFN-alpha. CONCLUSIONS: Weaker signals for Crk and Daxx may participate in less megakaryocyte suppressive activity of IFN-zeta/limitin and may distinguish IFN-zeta/limitin from IFN-alpha in megakaryocytes. Our results extend the understanding about thrombocytopenia in patients with IFN-alpha treatment as well as the possibility for the clinical application of human homologue of IFN-zeta/limitin or an engineered cytokine with useful features of the IFN-zeta/limitin structure. PMID- 15781342 TI - The historical importance of the hand in advancing the study of human anatomy. AB - Anatomy is often considered the foundation of medical science and is basic to the education of health professionals. This report will attempt to summarize the history of anatomy from antiquity to modern times. In addition it will highlight the historical importance of the upper extremity and its relationship to the development of human anatomy as a recognized medical science. PMID- 15781343 TI - Tendon healing in vitro: modification of tenocytes with exogenous vascular endothelial growth factor gene increases expression of transforming growth factor beta but minimally affects expression of collagen genes. AB - PURPOSE: It is not clear how the transfer of exogenous growth factor genes to tenocytes affects collagen production. An increase in collagen production enhances the repair but an increase in growth factors that stimulate tissue fibrosis may cause adhesion. Gene therapy is a new way to regulate tendon healing but it has been explored rarely. We genetically modified tenocytes with the vascular endothelial growth factor (VEGF) gene and investigated the expression of the genes for collagen production in an in vitro model of the proliferating tenocytes. METHODS: Tenocytes were obtained from cultures of rat intrasynovial tendons and distributed randomly to 25 dishes. The tenocytes in the experimental dishes (n = 9) were treated for 12 hours with plasmid containing the VEGF complementary deoxyribonucleic acid and then were cultured for 5 days; the tenocytes in the control dishes (n = 8) did not receive the exogenous gene. Tenocytes in the other dishes received exogenous platelet-derived growth factor (PDGF) gene for comparison of the effects of VEGF gene therapy. Efficiency of the gene transfer was evaluated by presence of the transgene in the tenocytes which was detected by reverse transcription polymerase chain reactions. Levels of expression of types I and III collagen and transforming growth factor (TGF)-beta genes were determined by quantitative analysis of the products of reverse transcription polymerase chain reactions. RESULTS: Expression of the TGF-beta gene increased significantly in the cells treated with exogenous VEGF cDNA. Expression of type I and III collagen genes by tenocytes was affected minimally by transfer of the VEGF gene to the tenocytes and was significantly weaker than that stimulated by PDGF gene therapy. Efficient gene transfer was confirmed by the presence of the VEGF complementary deoxyribonucleic acid in the tenocytes receiving the transferred gene. CONCLUSIONS: Transfer of exogenous VEGF gene has very limited effects on the promotion of collagen production in the proliferating tenocytes. This study suggests that VEGF gene therapy is not as beneficial as PDGF gene therapy to tendon healing and may increase the activities of TGF-beta that are associated with adhesion formations. PMID- 15781344 TI - Optimizing independent finger flexion with zone V flexor repairs using the Massachusetts General Hospital flexor tenorrhaphy and early protected active motion. AB - PURPOSE: Independent FDS action has been cited to be problematic with repair of multiple tendons in zone V owing to adhesion formation between the flexor digitorum superficialis (FDS) and the flexor digitorum profundus (FDP) tendons. Of the several described flexor repair techniques the ideal tendon repair should be strong enough to allow for early active motion to minimize adhesion formation and maximize tendon healing. Biomechanical studies have proven the Massachusetts General Hospital (MGH) repair to be strong enough to allow for early active motion. The purpose of this study was to examine the use of the MGH technique for zone V flexor tendon injuries to allow for early protected active motion to achieve independent finger flexion through better differential gliding of the tendons. METHODS: We performed a retrospective review 168 zone V finger flexor tendon repairs for 29 patients performed consecutively over 4 years when early active motion was not contraindicated. The same early protected active motion protocol was used for all of these patients. We reviewed total active motion, independent flexion, rupture, and need for tenolysis. These injuries involved 103 FDS and 65 FDP tendons to 103 fingers. The median follow-up period was 24 weeks. Of these 29 patients 19 were men and 10 were women. The average patient age was 28 years. RESULTS: The total active motion for these zone V repairs was 236 degrees +/- 5 degrees Overall 97 of 103 digits attained good to excellent function and 88 of 103 developed some differential glide. One of these patients required a tenolysis. Three repairs ruptured in 1 patient owing to suture breakage that was associated with noncompliance with the dorsal extension block splint. CONCLUSIONS: Our retrospective review of 168 consecutive flexor tendon repairs showed that the MGH technique allowed for early protected active motion, which provided good to excellent functional outcomes with 88 of 103 developing independent finger flexion at an acceptably low complication risk. PMID- 15781346 TI - Flexor digitorum profundus tendon to bone tunnel repair: a vascularization and histologic study in canines. AB - PURPOSE: Recent in vivo canine studies have shown incomplete restoration of the flexor digitorum profundus (FDP) insertion site after transection and repair to the cortical surface of the distal phalanx. Previous biomechanical analyses of tendon to bone surface repair have suggested that repair site gap formation of greater than 3 mm occurs frequently under physiologic loads. A recent ex vivo investigation into a novel repair of the FDP tendon into a bone tunnel in the distal phalanx showed improved tensile properties with a decrease in repair site gap formation. Time-zero data, however, do not always accurately reflect in vivo responses. The repair response of the FDP tendon when placed in an osseous compartment is not known. The purpose of this study was to analyze the histologic and vascular anatomic properties of the FDP insertion site after transection and repair in a bone tunnel within the distal phalanx. METHODS: Twenty-six FDP tendon to bone repairs were performed in 13 adult mongrel dogs after insertion site transection. The tendons were repaired in a bone tunnel in the distal phalanx. Vascular analysis of the tendon and repair site was performed by using a modified Spalteholtz technique and routine hematoxylin-eosin staining was used to assess histologic properties of the repair. RESULTS: In normal specimens the vascular analysis showed that there was a distal network of vessels extending 1- to 2-cm proximal to the FDP insertion site. At 10 days after repair the distal tendon segment tendon remained avascular. By 21 days after repair there was proximal migration of an unorganized reticular network of tendon surface vessels with sparse intratendinous communications. At 6 weeks after repair the structure of the distal tendon vascular network resembled that of normals. The vascular response of the tendon within the bone tunnel followed a similar time frame. Histologic analysis showed an inflammatory reaction in the bone tunnel leading to a progressive degradation of that portion of the FDP tendon that resided in the tunnel. Tendon necrosis was not seen. CONCLUSIONS: The FDP tendon, after insertion site transection and repair in a bone tunnel, undergoes a process of neovascularization and revascularization over a period of 6 weeks. There is a progressive loss of tendon parenchyma within the bone tunnel and the suture tracks appeared to serve as conduits for the ingrowth of inflammatory tissue. Restoration of the normal 4-zone tendon-bone interface was not seen. Although ex vivo biomechanical assessment of tendon repair in a bone tunnel appears promising, the repair response in vivo may not be favorable for tendon to bone healing. The progressive tendon degeneration that was observed here may have detrimental effects on repair site tensile properties, increasing the potential for early failure. PMID- 15781345 TI - The biomechanical analysis of a tendon fixation device for flexor tendon repair. AB - PURPOSE: Stainless steel suture is high in tensile strength but is not widely used in flexor tendon repair because of difficulty with handling and knot tying. The purpose of this study was to examine the biomechanical characteristics of the single-strand multifilament stainless steel Teno Fix device (Ortheon Medical, Winter Park, FL) designed for zone II flexor digitorum profundus (FDP) tendon repair. METHODS: Sixty cadaveric flexor tendons were transected and randomized to receive a Teno Fix or 4-stranded (3-0 or 4-0 braided polyester) suture repair; all repairs were tested with and without a 5-0 monofilament polypropylene circumferential epitendinous suture. By using a material testing system all tendons were tested to failure in tension using a linear model with a loading rate of 1 mm/s. Stiffness, force, and energy at both 2-mm gap and peak force were calculated from the resulting force-displacement curves. RESULTS: The 2-mm gapping force was significantly greater for the Teno Fix and the 3-0 repairs than for the 4-0 repairs. The energy absorbed up to 2-mm gap was significantly greater for the Teno Fix, however, than for all suture repairs both with and without a circumferential suture. There was no statistically significant difference in peak force or energy absorbed at peak force between the Teno Fix and suture repairs; the average gap at peak force for all repairs was 5.2 mm. The addition of a circumferential suture increased the 2-mm gapping and peak forces of the Teno Fix repair to 54.5 N and 66.7 N, respectively. CONCLUSIONS: Increased strength and energy absorbed at 2-mm gap and ease of installation makes the Teno Fix a promising repair method. PMID- 15781348 TI - Functional outcome of extensor carpi radialis longus transfer for finger flexion in posttraumatic flexor muscle loss. AB - PURPOSE: The purpose of this study was to assess the functional outcome after extensor carpi radialis longus (ECRL) transfer for restoration of finger flexion in patients with flexor muscle loss after direct trauma. METHODS: We evaluated 8 patients who had ECRL transfer between 1995 and 2003. Flexion gained was assessed by measuring the digit-to-palm distance (DPD). The grip strength was compared with that of the opposite normal limb. The average follow-up period was 41 months. We compared the results obtained with other modalities of restoration of finger flexion, namely a pedicled latissimus dorsi muscle transfer or a free functioning muscle transfer (FFMT) using the series available in the literature. RESULTS: Four patients had a good result with a DPD of 0 cm in all fingers and an average grip strength of 65% of the opposite hand. Two patients had an average result with a DPD of 1.5, 2, 1.7, and 1.5 cm for the index, middle, ring, and small fingers, respectively, and an average grip strength of 58%; 2 patients had a poor result with a DPD of 5.0, 5.5, 5.0, and 3.0 cm for the index, middle, ring, and small fingers, respectively, and with an average grip strength of 21% of the opposite hand. CONCLUSIONS: The ECRL transfer yields good results if the intrinsic muscles of the hand are functioning, the extensor compartment is uninjured, and the lower third of the forearm where the tendon junction is performed is relatively unscarred. In such instances the range of movement and grip strength achieved are better than a latissimus dorsi muscle pedicle graft and are comparable with a FFMT. This is achieved earlier than the time taken for reinnervation of FFMT and without the attendant risks for flap failure. The ECRL transfer for finger flexor restoration is a more simple alternative that should be considered when possible. PMID- 15781347 TI - Zone I flexor digitorum profundus repair: an ex vivo biomechanical analysis of tendon to bone repair in cadavera. AB - PURPOSE: Biomechanical studies of standard flexor digitorum profundus (FDP) tendon to bone repairs show ultimate strengths greater than the applied loads of early motion rehabilitation protocols. Strain data, however, indicate the potential for significant repair site gapping under these physiologic loads. Gaps in excess of 3 mm have been shown to prevent the time accrual of strength in midsubstance tendon repairs and may prevent the restoration of the normal architecture of the tendon-bone interface. Improving the time-zero tensile properties of FDP insertion site repairs may help obviate these issues and improve clinical outcomes. The purpose of this study was to evaluate the ex vivo biomechanical properties of 2 new repair techniques in comparison with the standard FDP tendon to distal phalanx cortical surface repair. METHODS: Thirty human cadaver FDP tendons were released from their insertion sites by sharp dissection and repaired to bone using 1 of 3 repair techniques. Load to failure testing was performed with a servohydraulic materials-testing system (model 8500R; Instron, Canton, MA) analyzing ultimate force, strain at 20 N, rigidity, force to 2-mm gap formation, and displacement at failure. RESULTS: The results of the failure tests indicate that repairs performed with the addition of a peripheral suture had a greater ultimate force, had increased resistance to gap formation, and had increased rigidity and decreased strain at 20 N compared with the tunnel-only and volar cortical surface to the distal phalanx repairs. Although there were no statistically significant differences in ultimate force or rigidity between the tunnel-only and volar cortical surface repairs, the tunnel only repairs showed lower strain values and increased values for resistance to 2 mm gap formation when compared with the volar cortical surface repairs. There were no differences among any of the repair groups with regard to the magnitude of tendon displacement from the repair site at failure. CONCLUSIONS: The addition of a peripheral suture to the FDP tendon to bone tunnel repair construct improves the time-zero tensile properties as evidenced by statistically significant increases in ultimate force, rigidity, and resistance to gap formations of 2 mm. In comparison with a volar cortical surface repair, the bone tunnel-only repairs were effective at decreasing the amount of repair site strain during applied loads of 20 N. If these improved time-zero tensile properties persist during the early stages of healing, they may help decrease the incidence of repair-site gap formation associated with the forces of early motion rehabilitation protocols. PMID- 15781349 TI - Biomechanical properties of the brachioradialis muscle: Implications for surgical tendon transfer. AB - PURPOSE: To understand the mechanical properties of the brachioradialis (BR) muscle and to use this information to simulate a BR-to-flexor pollicis longus (FPL) tendon transfer for restoration of lateral pinch. METHODS: The BR mechanical properties were measured intraoperatively. Passive elastic properties were measured by elongating BR muscles at constant velocity while they were attached directly to a dual-mode servomotor. Sarcomere length was measured intraoperatively and in situ by laser diffraction with the elbow fully extended. Then both the mechanical and structural properties were programmed into a surgical simulator to test the hand surgeon's decision making when tensioning muscles in a simulated BR-to-FPL tendon transfer. RESULTS: Passive mechanical BR properties were highly nonlinear. Under slack conditions sarcomere length (mean +/- standard deviation) was 2.81 +/- 0.10 microm (n = 4), corresponding to an active force of 93% maximum. Sarcomere length of the BR measured in situ with the elbow fully extended and the forearm in neutral rotation was 3.90 +/- 0.27 microm (n = 8), corresponding to an active force of only 23% maximum. Surgeons, who tensioned the BR for transfer into the FPL using only tactile feedback from the surgical simulator, attached the muscle at a passive tension of 5.87 +/- 0.97 N, which corresponded to a sarcomere length of 3.84 microm and an active muscle force of 27% maximum. Passive BR tension when both tactile and visual information were provided to the surgeon was significantly lower (2.42 +/- 0.72 N), corresponding to a sarcomere length of 3.56 mum and a much higher active muscle force of 45% maximum. CONCLUSIONS: When these data were used to model pretransfer and posttransfer function dramatic differences in predicted function were obtained depending on the tensioning protocol chosen. This emphasizes the point that the decision-making process used during muscle tensioning has a profound effect on the functional outcome of the transfer. PMID- 15781350 TI - Adequacy of laboratory simulation of in-line skater falls. AB - PURPOSE: Currently most laboratory simulation of distal radius fractures and wrist injuries has been with axial limb loading or vertical drop technique. To better assess the contribution of horizontal velocity in momentum in the development of wrist injury and wrist fracture we have compared forces and fracture patterns for fall simulations involving strictly vertical impacts with those that incorporated horizontal momentum in fresh-frozen upper extremities. METHODS: The premise for testing was based on a forward fall onto the palmar surface of an outstretched arm. A 45 degrees-incline impact device was used to model the horizontal and vertical velocity components of a skater fall at a representative speed. Sled mass was 26.2 kg. A 45.5-kg vertical impact system furnished contrasting data and represented prior state-of-the-art technology. Drop heights were adjusted to compensate for discrepancy in sled mass. Measurements from either impact system included vertical and horizontal forces and kinetic energy at impact. Fracture patterns were assessed radiographically. Nine trials were conducted with the incline device and 11 trials were conducted with the vertical impact system. RESULTS: The fracture rate was substantially higher for vertical impacts versus incline impacts (82% vs 33%). The rate of carpal fractures also was higher. Vertical and horizontal forces were similar statistically although slightly greater for vertical impacts. Kinetic energy, however, was 3 times greater for incline impacts than for vertical impacts. These kinetic parameters support the argument that the incline impacts should have resulted in far greater injury. CONCLUSIONS: The data suggest horizontal momentum changes limb loading. We submit that published methods for modeling skater falls have failed to provide a true simulation of the event in question. PMID- 15781351 TI - Functional outcome of unstable distal radius fractures: ORIF with a volar fixed angle tine plate versus external fixation. AB - PURPOSE: The purpose of this study was to compare the outcomes of 2 treatments for unstable distal radius fractures: open reduction internal fixation (ORIF) through a volar approach with a fixed-angle implant and a standard external fixation (EF) method. METHODS: This study included patients with comminuted unstable intra-articular and extra-articular distal radius fractures treated by a single surgeon. Data were gathered retrospectively on 11 patients treated with EF who had been followed up for an average of 47 months (range, 12-84 mo). Prospective data were gathered on 21 patients who were treated with ORIF through a volar approach with a fixed-angle implant. Follow-up evaluation for this group averaged 17 months (range, 12-24 mo). The 2 groups were compared for range of motion (ROM), strength, and functional outcome as measured by the Patient Rated Wrist Evaluation (PRWE) and the Disability of the Arm, Shoulder, and Hand Questionnaire (DASH). Fracture reduction was evaluated from radiographs taken at the last follow-up visit and compared between groups. RESULTS: The mean passive wrist ROM at the final follow-up evaluation in EF patients was 59 degrees extension and 57 degrees flexion, compared with 63 degrees extension and 64 degrees flexion in patients treated with ORIF. Passive pronation/supination arc of motion was similar for the 2 groups, as were the DASH and PRWE scores. Grip strength as a percentage of the opposite wrist was significantly greater in the external fixation group, a possible consequence of longer follow-up evaluation. Final radiographic measurements for the EF group averaged 5 degrees volar tilt and 25 degrees radial inclination, with 2.2-mm ulnar-positive variance. The ORIF with volar plating group averaged 10 degrees volar tilt and 22 degrees radial inclination, with .5-mm ulnar-negative variance. Radial length and volar tilt were significantly greater for the ORIF group. The average final intra-articular step-off was significantly different, with 1.4-mm step-off in the EF group and .4 mm in the ORIF group. CONCLUSIONS: The use of ORIF with a volar fixed-angle implant resulted in stable fixation of the distal articular fragments, allowing early postsurgical wrist motion. The PRWE and DASH scores for the groups were equivalent, whereas intra-articular step-off, volar tilt, and radial length were better in the ORIF group. There were few complications, implant removal was not necessary, and early postsurgical wrist ROM was initiated without loss of reduction. PMID- 15781352 TI - The effects of implant composition on extensor tenosynovitis in a canine distal radius fracture model. AB - PURPOSE: Dorsal plating of distal radius fractures with titanium plates has resulted in clinically observed tenosynovitis and tendon rupture. The goal of this study was to investigate whether titanium-based implants result in more extensor tendon inflammation than matched stainless-steel implants in a canine fracture model. METHODS: An osteotomy was created in the distal radius of 18 beagles and fixed with 2.7-mm 4-hole plates composed of commercially pure titanium, titanium alloy (Ti-Al6-V4), or 316L stainless steel. Animals were killed at an average of 4 months. Tendon gliding was assessed by applying a force at the extensor musculotendinous junction and noting gliding. Histologic grading (mild, moderate, severe) was based on cellular hypertrophy, hyperplasia, and leukocytic infiltration. RESULTS: Tendons glided freely in 100% stainless-steel specimens, 75% of titanium alloy, and 43% of commercially pure titanium groups. A severe inflammatory reaction was identified in 60% of the titanium alloy (Ti-A16 V4) group, 57% of the pure titanium group, and 0% of the stainless-steel group. CONCLUSIONS: Dorsal plating of the canine radius with commercially pure titanium or titanium alloy implants produced a greater inflammatory peritendinous response than matched stainless-steel implants. PMID- 15781353 TI - Entrapment of the flexor digitorum superficials in the radius fracture site. AB - Entrapment of the flexor tendon after fracture of both forearm bones is very rare. This report describes entrapment of the middle finger flexor digitorum superficialis (FDS) tendon alone in the radius fracture site. A 13-year-old boy fractured both forearms and had closed reduction and a long-arm cast. After the cast was removed he had full middle finger motion with the wrist flexed but was unable to extend the proximal interphalangeal or metacarpophalangeal joints and could extend only the distal interphalangeal joint of the middle finger with the wrist in the neutral or extension position. Entrapment of the middle finger FDS tendon was suspected. After surgical release of the FDS tendon at the fracture site the patient had good functional results. PMID- 15781354 TI - Structural properties of reconstruction constructs for the interosseous ligament of the forearm. AB - PURPOSE: The Essex-Lopresti fracture-dislocation, also termed longitudinal radioulnar dissociation (LRUD), results in major functional impairment from pain and limitation of motion at the wrist and elbow. Interosseous ligament (IOL) reconstruction has been proposed to help treat LRUD and restore forearm stability. The objective of this study was to evaluate the biomechanical structural properties of 3 different IOL reconstruction constructs and of the intact IOL for comparison. METHODS: Structural tensile testing was performed on 24 fresh-frozen cadaveric forearms with 4 different forearm conditions: IOL intact and IOL reconstructed with Achilles tendon, flexor carpi radialis (FCR) tendon, and bone-patellar tendon-bone. Isolated radius-IOL-ulna constructs were loaded to failure in a materials testing machine with force applied along the local fiber direction. RESULTS: Stiffness in the intact IOL was 129 +/- 31 N/mm, which was significantly stiffer than any of the constructs tested. The intact IOL was 8 times stiffer than the Achilles tendon construct, 7 times stiffer than the FCR construct, and 3 times stiffer than the bone-patellar tendon-bone construct. The Achilles tendon and FCR constructs were similar to each other biomechanically but the bone-patellar tendon-bone construct was slightly stiffer than the Achilles tendon and FCR constructs. CONCLUSIONS: All graft constructs tested were inferior structurally to the intact IOL. The results of this study provide a biomechanical basis for graft selection for reconstruction of the IOL. PMID- 15781355 TI - Reconstruction of the interosseous ligament restores normal forearm compressive load transfer in cadavers. AB - PURPOSE: Longitudinal radioulnar dissociation may result when compressive load to the hand causes excessive proximal migration of the radius with interosseous ligament (IOL) disruption and radial head fracture. Although radial head salvage or arthroplasty and temporary distal radioulnar joint pinning constitute the current treatment for this injury IOL reconstruction has been proposed to restore normal forearm mechanics. To help provide a biomechanical basis for IOL reconstruction we measured load transfer and proximal migration of the radius with the IOL intact, cut, and reconstructed while leaving the radial head intact. METHODS: We dissected the central third of 12 normal cadaveric forearms to bone IOL-bone. We applied 136 N of compressive load to the hand and measured proximal radial migration and 3-dimensional force vectors acting in the distal radius, distal ulna, IOL, proximal radius, and proximal ulna. Experiments were performed in neutral forearm rotation and neutral elbow varus-valgus. The protocol was repeated with the IOL intact, cut, and reconstructed with single and double flexor carpi radialis allografts. RESULTS: With the IOL intact 94% +/- 3 % of hand load was in the distal radius with 75% +/- 2% was transmitted to the proximal radius. With the IOL cut 92% +/- 2% of hand load was in the distal radius and this was unchanged at the proximal radius. With single flexor carpi radialis reconstruction 94% +/- 3% of hand load was in the distal radius and 80% +/- 2% was in the proximal radius; with double flexor carpi radialis reconstruction these numbers changed to 95% +/- 3% and 74% +/- 2%, respectively. Forces in the forearm were mainly in the longitudinal and transverse directions with negligible components in the dorsal-volar direction. Transverse force in the IOL was about half that of the longitudinal force in the intact and reconstructed states. CONCLUSIONS: When the IOL is cut and load is applied to the hand the unloading of the proximal radius and the transversely directed force that compresses normally across the proximal and distalradioulnar joints is lost. Reconstruction of the IOL can restore the normal load transfer characteristics. These data help provide a biomechanical basis for IOL reconstruction. Further research on the biomechanics of IOL reconstruction in the setting of radial head replacement along with clinical evaluation is needed. PMID- 15781357 TI - Graft reconstruction of the interosseous membrane in conjunction with metallic radial head replacement: a cadaveric study. AB - PURPOSE: Longitudinal radioulnar dissociation (Essex-Lopresti injury) occurs when traumatic axial loading through the wrist disrupts the interosseous membrane (IOM) of the forearm and fractures the radial head. Proximal migration of the radius results in an ulnar-positive wrist, which can lead to painful ulnar-sided wrist degeneration and distal radioulnar joint instability. The purpose of this study was to measure the ability of an IOM reconstruction used in combination with a metal prosthetic radial head implant to reduce distal ulnar forces in a cadaveric model. The effects of varying the initial graft pretension on distal ulnar force were also studied. METHODS: Twelve fresh frozen and thawed cadaveric forearms had a miniature load cell installed to record force in the distal ulna as the wrist was loaded axially to 134 N of compression force in neutral rotation. Intact forearms were tested first with the elbow in valgus and varus alignments. Loading tests were repeated after (1) insertion of a metal radial head implant that restored radius anatomic length, (2) excision of the IOM (with a radial head implant), and (3) reconstruction of the IOM using a palmaris longus tendon autograft (with a radial head implant). The implant then was removed and loading tests were repeated using 3 levels of initial graft pretension. RESULTS: Mean distal ulnar forces with an intact forearm were 23% of applied wrist force in the varus alignment and 12% in the valgus alignment. Mean force levels after insertion of the implant were 18% (varus) and 13% (valgus); these were not significantly different from corresponding values for the intact forearm. Mean force levels after section of the IOM were 30% (varus) and 14% (valgus); these were not significantly different from corresponding values for the intact forearm (varus and valgus) but the mean for varus was significantly greater than the corresponding value with an implant. After IOM reconstruction with a palmaris longus tendon tensioned to 22 N mean distal ulnar forces were 8% (varus) and 7% (valgus); these means were significantly less than the corresponding values for all prior test conditions. With the radial head removed increasing the level of graft pretension reduced significantly mean distal ulnar force. CONCLUSIONS: With the IOM resected insertion of a metal radial head implant alone did not reduce distal ulnar forces to intact forearm levels. When an IOM reconstruction was performed in combination with the implant mean distal ulnar force was reduced significantly to a level below that for the intact forearm. Applying pretension to the graft displaced the radius distally thereby making the wrist more ulnar negative and reducing distal ulnar force. Our results suggest that an IOM reconstruction used in combination with a metal radial head implant theoretically could help reduce distal ulnar impaction in an Essex-Lopresti injury. PMID- 15781356 TI - Reconstruction of the interosseous membrane of the forearm with a graft substitute: a cadaveric study. AB - PURPOSE: Longitudinal radioulnar dissociation occurs when traumatic axial loading through the wrist disrupts the interosseous membrane (IOM) of the forearm and fractures the radial head (Essex-Lopresti injury). Proximal migration of the radius results in a wrist with a positive ulnar variance, which leads ultimately to painful ulnar-sided wrist degeneration and wrist pain during grasping activities that involve axial loading or ulnar deviation of the wrist. In theory reconstruction of the IOM with a graft substitute can limit proximal migration of the radius, thereby preserving wrist function. The objective of this study was to measure the abilities of 3 graft tissues to limit proximal radial displacement compared with the native IOM in a radial head-deficient cadaver model. METHODS: Sixteen fresh-frozen cadaveric forearms were loaded axially to 134 N through the potted central 3 metacarpals; the elbow was flexed to 90 degrees with the wrist in neutral rotation. Proximal displacement of the radius relative to the capitellum was measured. With the radial head excised specimens were first tested with the IOM intact. The IOM was then sectioned and central band IOM reconstructions were performed on each specimen using the following tissues: palmaris longus tendon, flexor carpi radialis (FCR) tendon, and a 1-cm- wide bone patellar tendon-bone (BPTB) onlay allograft. Ten loading cycles were performed with each test configuration. Proximal radial displacement between 13.4 N and 134 N of applied wrist force was analyzed for the 10th loading cycle. The increase in proximal displacement between the first and 10th loading cycles (recorded at 134 N of wrist force) represented permanent elongation of the graft. RESULTS: Mean cross-sectional areas were 5.11 mm2 for the palmaris longus tendon, 15.23 mm2 for the FCR tendon, and 51.59 mm2 for the BPTB allograft. Mean proximal radial displacements were 3.04 mm (intact IOM), 4.37 mm (BPTB), 4.92 mm (FCR tendon), and 6.43 mm (palmaris tendon); all means were significantly different from each other. Mean permanent graft elongations were 0.06 mm (IOM), 0.36 mm (BPTB), 1.25 mm (FCR tendon), and 1.80 mm (palmaris tendon); all means were significantly different from each other with the exception of means for palmaris longus vs FCR and BPTB vs IOM. CONCLUSIONS: No graft reconstruction limited proximal radial displacement as effectively as the native IOM. Of the 3 graft tissues tested the BPTB allograft had the greatest cross-sectional area, allowed the least proximal radial displacement, and displayed the least permanent elongation after 10 cycles of loading. The relatively thin and narrow palmaris longus tendon appears to be the least desirable choice for IOM reconstruction because of its relatively low stiffness and tendency to elongate permanently after cyclic loading. When the radial head is absent rupture of the IOM allows unopposed proximal displacement of the radius relative to the ulna as the wrist is loaded axially. In the present tests all 3 graft tissues used to reconstruct the IOM limited proximal radial displacement. The choice of graft material is an important variable if IOM reconstruction is considered for treatment of an Essex-Lopresti injury. PMID- 15781358 TI - Patterns of subchondral bone mineralization in the distal radioulnar joint. AB - PURPOSE: Studies have shown that it is possible to derive direct knowledge about the actual mechanical conditions of the wrist by analyzing the subchondral mineralization. The aim of the present study was to evaluate the distribution of the subchondral bone mineralization of the distal radioulnar joint (DRUJ) noninvasively in living subjects by using computed tomography (CT) osteoabsorptiometry to gain new information about the long-term loading conditions. METHODS: Twenty-two wrist joints were investigated in 11 healthy young subjects by means of CT osteoabsorptiometry. The CT scans of the DRUJ were taken in the axial plane in neutral position of the forearm and in both maximum pronation and supination. The CT datasets of 1.5-mm sections were obtained and then transferred to an image-analyzing system. The subchondral bone plate in each section was isolated, reconstructed in 3 dimensions, and converted into a false color series. RESULTS: The maximum subchondral bone density in the sigmoid notch of the radius was found along the distal border in all wrists. It was located dorsally in 10 wrists, palmarly in 8 wrists, and centrally in 4 wrists. The maximum bone density on the corresponding articular surface of the ulna was found dorsally in 10 cases, centrally in 8 cases, and palmarly in 4 cases. In 13 cases the maximum bone density was found in direct opposition on radius and ulna in neutral position. A statistically significant difference could not be detected in subjects with an ulna minus variance nor in those with a DRUJ angulation greater than 10 degrees . CONCLUSIONS: Our results show that in the sigmoid notch the load is transmitted through either the dorsal or palmar parts of the joint. In contrast the maximum bone density on the side of the ulna was found dorsally and centrally. We conclude that the ulna receives the maximum load in neutral position and supination, whereas because of the dorsopalmar translation of the ulnar head the radius may lead the ulna with either its palmar or dorsal borders during pronosupination. The ligamentous apparatus, the shape of the joint, and the ulna variance, however, may influence load transmission. PMID- 15781359 TI - Effects of wafer resection and hemiresection from the distal ulna on load-sharing at the wrist: a cadaveric study. AB - PURPOSE: Ulnar impaction syndrome is commonly the result of a naturally occurring ulnar-positive condition, distal radius fracture malunion, or collapse of a fractured radial head. The Feldon wafer procedure and the Bowers distal hemiresection procedure are designed to decrease force transmitted through the distal ulna. The purpose of this study was to measure the effects of these procedures on distal ulnar loading with varying degrees of ulnar positivity at the wrist. METHODS: Using a specially designed miniature load cell distal ulnar force was measured in 20 fresh-frozen cadaveric forearms as the wrist was loaded axially to 134 N in neutral forearm rotation; tests were performed in valgus alignment with the elbow flexed to 90 degrees . Ulnar positivity (0 mm, +2 mm, +4 mm, and +6 mm) was created by incremental shortening of the distal radius using a sliding plate. The radial neck was sectioned transversely and the radial head fragment was fixed in its anatomic position using cemented metal prongs connected to a rigid bar; this allowed the radial head to be disconnected and tilted out of the loading pathway to simulate an excised radial head. Loading tests (with and without the radial head in place) were repeated after removal of a 3-mm wafer of bone beneath the triangular fibrocartilage complex, and again after a hemiresection of the distal ulna to the base of the ulnar styloid process. RESULTS: For each ulnar status condition (intact, wafer removal, hemiresection) the mean distal ulnar force generally increased as the wrist became more ulnar positive. Both wafer removal and hemiresection significantly decreased mean distal ulnar forces under all conditions of ulnar variance, with or without the radial head in place. With the radial head in place the mean distal ulnar forces (expressed as a percentage of applied wrist force) for the 0-mm condition were 16.9% (intact), 3.8% (wafer removal), and 3.5% (hemiresection); corresponding values for the +6-mm condition were 61.6% (intact), and 39.8% (wafer removal), 15.1% (hemiresection). With the radial head removed the mean distal ulnar forces for the 0-mm condition were 31.7% (intact), 4.6% (wafer removal), and 4.4% (hemiresection); corresponding values for the +6-mm condition were 96.4% (intact), 71.6%, (wafer removal), and 27.2% (hemiresection). The decrease of distal ulnar force resulting from hemiresection was significantly greater than that for wafer removal for all ulnar-positive conditions; force reductions were not significantly different between the 2 procedures with neutral ulnar variance. CONCLUSIONS: The results of the present study can help to offer a biomechanical basis for choosing between a Feldon wafer procedure and a Bowers hemiresection procedure in patients with ulnocarpal impaction syndrome. Both procedures produced equal decreases of distal ulnar force in the intact forearm. With an ulnar-positive wrist, the condition for which the procedure would be performed commonly, the hemiresection was more effective in decreasing force transmitted through the distal ulna. PMID- 15781360 TI - Patterns of bone defect in scaphoid nonunion: a 3-dimensional and quantitative analysis. AB - PURPOSE: To clarify patterns and size of bone defect in scaphoid nonunion in order to facilitate accurate correction of scaphoid deformity. METHODS: Three dimensional computed tomography was used to examine 24 patients with scaphoid nonunion. Configuration and size of bone defect were quantified and computed on the basis of fracture location. Cases were categorized as distal or proximal based on location of the fracture line relative to the dorsal apex of the scaphoid ridge. RESULTS: Distal scaphoid fractures displayed wedge-shaped bone defects with the base facing volarly. Proximal scaphoid fractures exhibited flat, crescent-shaped defects that presented only around the fracture site. The size of bone defects was significantly greater for distal fractures than for proximal fractures. CONCLUSIONS: Whether the fracture line passes distal or proximal to the dorsal apex of the scaphoid ridge is a crucial factor in the generation of bone defect. A large wedge-shaped bone graft from the volar side is necessary for distal nonunion whereas a small cancellous bone graft from the dorsal side may be preferable for proximal nonunion. PMID- 15781361 TI - Acute coronal plane scaphoid fracture and scapholunate dissociation from an axial load: a case report. AB - Coronal fractures of the scaphoid are rare and can be difficult to diagnose. Axial load injuries that result in a complete coronal fracture of the scaphoid associated with an acute scapholunate dissociation are exceedingly rare. In our patient the radiographic finding of wide scapholunate dissociation was obvious; however, the coronal scaphoid fracture was not recognized initially nor suspected. During surgery the coronal scaphoid fracture was identified, reduced anatomically, and fixed with a compression screw. The scapholunate ligament also was repaired. A good result was obtained with return to sports with extension of 60 degrees and flexion of 70 degrees , grip strength equal to that of the uninjured wrist, and no radiographic problems (arthrosis, avascular necrosis, nonunion). PMID- 15781362 TI - Thumb carpometacarpal arthroscopy: a topographic, anatomic study of the thenar portal. AB - Arthroscopic treatment of osteoarthritis of the thumb carpometacarpal joint has been well described; however, the current site of the 2 working portals, especially the 1-R portal, may be complicated by neurovascular morbidity. This is owing to the close proximity of these portals to important nerves and vessels surrounding the carpometacarpal joint. We studied 7 cadaveric wrists to map out the topographic, anatomic, and arthroscopic position of a new thenar portal. We hypothesized that it would increase considerably the safe zone distances of the portal from vital structures of the joint compared with the traditional portals. This study showed that the thenar portal increases considerably the safe zone of the portal from the sensory branches of the radial nerve and the radial artery and does not put the motor branch of the median nerve at risk. In addition the thenar portal allows for better visualization of the carpometacarpal joint, which leads to improved ability to perform arthroscopic trapeziectomy. PMID- 15781363 TI - Results from a degradable TMC joint Spacer (Artelon) compared with tendon arthroplasty. AB - PURPOSE: A new spacer for the trapeziometacarpal joint (TMC) based on a biological and tissue-preserving concept for the treatment of TMC osteoarthritis (OA) has been evaluated. The purpose was to combine a spacing effect with stabilization of the TMC joint. METHODS: Artelon (Artimplant AB, Sweden) TMC Spacer is synthesized of a degradable polyurethaneurea (Artelon), which has been shown to be biocompatible over time and currently is used in ligament augmentation procedures. Fibers of the polymer were woven into a T-shaped device in which the vertical portion separates the bone edges of the TMC joint and the horizontal portion stabilizes the joint. Fifteen patients with disabling pain and isolated TMC OA were included in the study. Ten patients received the spacer device and the remaining 5 (control group) were treated with a trapezium resection arthroplasty with abductor pollicis longus (APL) stabilization. The median ages of the 2 groups were 60 and 59 years, respectively. Pain, strength, stability, and range of motion were measured before and after surgery. Radiographic examination was performed in all patients before and after surgery. At follow-up evaluation 3 years after surgery an unbiased observer evaluated all patients. Biopsy specimens were obtained from 1 patient 6 months after surgery. RESULTS: All patients were stable clinically without signs of synovitis. In both groups all patients were pain free. The median values for both key pinch and tripod pinch increased compared with before surgery in the spacer group but not in the APL group. The biopsy examinations showed incorporation of the device in the surface of the adjacent bone and the surrounding connective tissue. No signs of foreign-body reaction were seen. CONCLUSIONS: This study showed significantly better pinch strength after Artelon TMC Spacer implantation into the TMC joint compared with APL arthroplasty. PMID- 15781364 TI - Use of volar plate for reconstructing the radial collateral ligament after metacarpophalangeal arthroplasty of fingers in rheumatoid arthritis: surgical technique. AB - We present a technique for reconstructing the radial collateral ligament using the volar plate after metacarpophalangeal arthroplasty of fingers in rheumatoid arthritis. This technique is indicated when the original collateral ligament is very attenuated and cannot be directly imbricated. The technique is surgically simple, has the advantage of using locally available tissue, and has produced results that are both clinically and radiologically satisfactory. PMID- 15781366 TI - Correction of pincer-nail deformities with autograft or homograft dermis: modified surgical technique. AB - The pincer-nail deformity is characterized by an excessively curved and distorted nail across the transverse dimension. Forty-nine sides (paronychial folds) were dissected off the distal phalanx periosteum with scissors and/or a small elevator. The dermis was placed between the paronychial fold and the plalanx to flatten the germinal and sterile matrix. Direct comparison of autograft dermis to homograft dermis did not show any significant differences in postcorrection appearance of the nail or relief of symptoms. Surgical time averaged 22 minutes less in those patients having reconstruction on both sides of one nail with homograft dermis. PMID- 15781365 TI - Reconstruction of the ulnar collateral ligament of the thumb metacarpophalangeal joint: a cadaver study. AB - PURPOSE: The purpose of this study was to compare the stiffness and strength of the native ulnar collateral ligament with 4 methods of static ulnar collateral ligament (UCL) reconstruction at the thumb metacarpophalangeal (MCP) joint. METHODS: Eleven fresh-frozen cadaver specimens were amputated at the carpometacarpal and interphalangeal joints and all soft tissues were removed except for the extensor pollicis brevis tendon, the proper and accessory collateral ligaments, and the volar plate. Each thumb metacarpal was potted in cement and the native UCL was loaded to failure at 30 degrees of MCP flexion. Ulnar collateral ligament reconstructions as described by Strandell, Osterman, Fairhurst, and a modification of the Glickel procedure then were performed. Each specimen was again loaded to failure and the moment at failure, stiffness, and angle at failure were calculated. RESULTS: None of the reconstructions duplicated the strength or stiffness of the native UCL. The modification of the Glickel procedure with interference knot fixation had a significantly higher moment at failure and was significantly stiffer than any of the other procedures. The differences in strength and stiffness between the Strandell, Osterman, and Fairhurst reconstructions were not statistically significant. There were no significant differences in angle at failure for any of the reconstructions. CONCLUSIONS: No static ligament reconstruction restores the normal stability characteristics of the thumb UCL. The anatomic reconstruction of the UCL with interference knot fixation of the tendon graft has far better strength and stiffness than any of the other reconstructions tested. These characteristics may allow for early motion at the MCP joint. PMID- 15781367 TI - Fibrohistiocytic tumor of the hand after high-pressure paintgun injury: 2 case reports. AB - Injections from high-pressure, airless paintguns are uncommon industrial accidents that may result in acute injuries, usually to the nondominant hand. These injuries commonly require emergent management including immediate surgical debridement to avoid tissue necrosis and loss of hand function. Rarely oleogranulomas and squamous cell carcinoma have been described as late complications. Herein we describe 2 patients who developed fibrohistiocytic tumors several months after a high-pressure paintgun injury. PMID- 15781369 TI - Percutaneous reduction of incipient malunion of phalangeal neck fractures. PMID- 15781370 TI - [Tissue engineering in the field of orthopaedics and traumatology]. PMID- 15781371 TI - [Use of growth factors in the repair of bone]. AB - Osteoformation is induced by numerous growth factors that play an important role in bone repair such as fracture healing. They may serve as therapeutic agent in the treatment of squeletal injuries in the orthopeadic and maxillo-facial fields. Among these proteins, Bone Morphogenetic Proteins (BMP) are the only known osteoinductive growth factors. Unfortunately, they are highly susceptible to proteolysis in vivo and require a suitable delivery system to potentiate their biological activity in a local, controlled and durable manner. In this aim, three options are under investigations: (i) osteoinductive materials made of appropriate carrier to release the protein in situ, (ii) in vivo gene therapy in which the gene is directly transfected in cells of the patient or (iii) ex vivo gene therapy in which cells are harvested from the patient, transfected with DNA in culture and then implanted in the defect. These different kinds of BMP delivery will be discussed. PMID- 15781372 TI - [Therapeutic application of mesenchymal stem cells in orthopaedics]. AB - Stem cell therapy of skeletal tissues involves the transplantation of stem cells to the tissues that have been damaged by injury or disease. Although these cells can be derived from embryos, the preferred source of skeletal stem cells is the bone marrow as it contains adult stem cells that can be easily driven towards a bone phenotype. More recently, cells with similar potentialities have also been derived from adipose tissue, muscle, or blood. A biomaterial (ceramics or polymers) is often required as a scaffold to promote cell adhesion, proliferation and differentiation as well as encourage vascular invasion and ultimately new bone formation. The first clinical studies are encouraging and suggests that stem cell therapy could be a prime method for bone reconstruction. PMID- 15781373 TI - UCB transplant supported by the co-infusion of a low number of highly purified CD34+ haploidentical cells: update of results. PMID- 15781374 TI - Ex vivo amplification of T cells from human cord blood. AB - The current status of immunotherapy for cancer is here summarized with particular attention to the new methodologies developed for ex vivo expansion of T cells from neonatal cord blood. Umbilical cord blood (UCB) mononuclear cells (MNC) generate CD45RA naive T lymphocytes when cultured under serum-deprived conditions with appropriate combinations of growth factors. These ex vivo generated T cells resemble precursors for the lymphoid lineage present in adult bone marrow in terms of active transcription of RAG-2 and pTalpha. PMID- 15781375 TI - Experiences with haploidentical stem cell transplantation in children with acute lymphoblastic leukemia. PMID- 15781376 TI - Biology and clinical applications of marrow mesenchymal stem cells. PMID- 15781377 TI - Unrelated donor umbilical cord blood transplantation in adults with hematologic malignancies. PMID- 15781378 TI - Unrelated donor transplant for acute leukaemia in children--the UK experience. PMID- 15781379 TI - Immunogenomics of hematopoietic stem cell transplantation. AB - Recipients of allogeneic hematopoietic stem cell transplantation (HSCT) incur the risk of graft-versus-host disease even when the donor is a sibling who shares the Major Histocompatibility Antigens. Therefore, even the perfect HLA match does not represent the optimal genetic match between donors and recipients in HSCT. In addition to the HLA complex other genetic systems operate and affect the outcome of HSCT. These include minor histocompatibility systems (Martin P. Applicability of matching for minor histocompatibility antigens in human bone marrow transplantation. In: Roopenian DC, Simpson E, editors. Minor histocompatibility antigens: From the laboratory to the clinic. Georgetown: Landis Bioscience; 2000. p. 97-103) (inducing bona fide allogeneic responses) as well as a series of functional polymorphisms in cytokines and chemokines and receptors genes (Transplantation 1997;64:553). Among the items affecting the outcome of HSCT the incidence and severity of infections have an important impact. Polymorphisms of genes controlling both arms of the immune responses to pathogens (innate versus cognate) are strong candidates for susceptibility factors to infection in allogeneic transplantation. These include the MHC alleles (HLA class I, class II, MIC) CD1, Toll and TLR genes MBP, MPO genes, ...). In addition to the NK alloreactivity induced by HLA class I epitopes mismatching (a common situation in HSCT) variations in the genotype of the KIR genes (Tissue Antigens 2001;57:358) may also be encountered between the donor and the recipient leading to potentially harmful or beneficial combinations. An integrated knowledge of the role and hierarchy of the most important genetic factors (MHC and non-MHC) will provide the rationale for a comprehensive matching in HSCT (Curr Opin Hematol 3 (1996) 416). This short review provides a panorama of this strategic issue for further development of HSCT. PMID- 15781380 TI - [Sodium/iodide symporter: physiopathological aspects and therapeutic perspectives]. AB - The existence of the natriuric/iodide symporter (NIS) represents a new view to understand the thyroid metabolism of iodide. Due to its cellular localisation on basolateral membrane, this transporter exerts an essential role in the biological functions of the thyroid, especially the capacity to accumulate iodide into the thyrocytes. Clinical perspectives of NIS activity modulation would ameliorate the diagnosis and the treatment of thyroid diseases by using radioisotopes transported by the NIS (131 iodide, 99m technetium, 188 rhenium). The study of the regulation pathways modulating the expression and the activity of the symporter NIS, would allow to understand pathogeny of benign or malignant diseases of the thyroid gland. The relative facility of the therapy management by 131 iodide and its good efficiency associated to the recent advance of NIS function also give an interesting perspective to the gene therapy treatment of the nonthyroid cancers despite existent methodological problems. PMID- 15781381 TI - [Angiotensin I-converting enzyme (ACE) for sarcoidosis diagnosis]. AB - Angiotensin I-converting enzyme (ACE) is synthesized by sarcoidotic granulomas defining this enzyme as a diagnosis and prognosis marker of sarcoidosis. This granulomatous disease, a frequent disease with unknown aetiology, is essentially localized in lung and skin, but many other localizations are possible, as well as in nervous system. The diagnosis is based on a panel of clinical, biological and radiological arguments. Serum ACE has a particular interest although its sensitivity and specificity for sarcoidosis are not perfect; ACE can also help to follow this disease either spontaneously or after corticoid therapy. ACE can be measured in bronchoalveolar lavage fluid (BAL) where it better signs the activity of the pulmonary disease than in serum. ACE in cerebrospinal fluid (CSF), and eventually in other biological fluids, allows the diagnosis of sarcoidosis with extra-pulmonary locations. The methods for ACE activity determination are essentially based on the use of polypeptidic synthetic substrates. By varying their nature and the conditions for use various assays were developed for serum, plasma, BAL, CSF and other fluids where ACE can help. The data must be given in a critical manner in function of the actual knowledge on ACE gene, the enzymatic properties of the protein and the limits of sensitivity and specificity of the assays sometimes reserved to peculiar biological fluids. PMID- 15781382 TI - Altering protein specificity: techniques and applications. AB - Protein engineering constitutes a powerful tool for generating novel proteins that serve as catalysts to induce selective chemical and biological transformations that would not otherwise be possible. Protocols that are commonly employed for altering the substrate specificity and selectivity profiles by mutating known enzymes include rational and random methods as well as techniques that entail evolution, selection and screening. Proteins identified by these techniques play important roles in a variety of industrial and medicinal applications and in the study of protein structure-function relationships. Herein we present a critical overview of methods for creating new functional proteins having altered specificity profiles and some practical case studies in which these techniques have been applied to solving problems in synthetic and medicinal chemistry and to elucidating enzyme function and biological pathways. PMID- 15781383 TI - Synthesis and biological study of a flavone acetic acid analogue containing an azido reporting group designed as a multifunctional binding site probe. AB - Flavone-8-acetic acid (FAA) is a potent immunomodulatory small molecule that is uniquely characterized as being active on mouse but not human cells. Although FAA is a potent inducer of murine cytokine, chemokine and interferon gene expression, its mode of action remains unknown. In this report, we describe the synthesis of a new flavone acetic acid (FAA) analogue, (2-[2-(4-azidophenyl)-4-oxochromen-8-yl ]acetic acid (compound 2). We demonstrate that compound 2 is equally active as the parent FAA in inducing chemokine gene expression and that the azide functional group is capable of reacting with a reporter molecule, such as the FLAG peptide-phosphine, under mild conditions. This reaction will be useful for detecting the drug-bound protein active complex utilizing an anti-FLAG antibody. PMID- 15781384 TI - Design, synthesis and characterization of a novel class of coumarin-based inhibitors of inducible nitric oxide synthase. AB - Inducible nitric oxide synthase (iNOS) has been implicated in various central and peripheral pathophysiological diseases. Our high throughput screening initially identified a weak inhibitor of iNOS, thiocoumarin 13. From this lead, a number of potent derivatives were prepared that demonstrate favorable potency, selectivity and kinetics. Compound 30 has an IC50 of 60 nM for mouse iNOS and 185-fold and 9 fold selectivity for bovine eNOS and rat nNOS, respectively. In cellular assays for iNOS, this compound has micromolar potency. Furthermore, two compounds (16 and 30) demonstrate a reasonable pharmacokinetic profile in rodents. The synthesis, SAR, and biological activity of this novel class of compounds is described. PMID- 15781385 TI - Isolation of brominated quinones showing chemiluminescence activity from luminous acorn worm, Ptychodera flava. AB - Luminous acorn worm, Ptychodera flava emits green light by stimulating with diluted hydrogen peroxide. We have recently reported isolation and structure determination of 2,3,5,6-tetrabromohydroquinone as a luminous substance and riboflavin as a possible light emitter. There are three other luminous substances in the extracts from P. flava, so here we report the isolation and structure determination of other luminous substances as 2,3,5-tribromohydroquinone, tetrabromo-1,4-benzoquinone, and 2,3,5-tribromo-6-(2,3,5-tribromo-4-hydroxy phenoxy)-benzene-1,4-diol. Besides, this is the first report of isolation of tetrabromo-1,4-benzoquinone from acorn worm. Structure-activity relationship of chemiluminescence activity of halogenated quinone derivatives reveals that a highly halogen substitution and 1,4-quinone skeleton are important for high chemiluminescence activity. PMID- 15781386 TI - Synthesis and biological evaluation of 3,4-diphenyl-1,2,5-oxadiazole-2-oxides and 3,4-diphenyl-1,2,5-oxadiazoles as potential hybrid COX-2 inhibitor/nitric oxide donor agents. AB - A group of 3,4-diphenyl-1,2,5-oxadiazole-2-oxides (3,4-diphenylfuroxans) and the corresponding N-desoxy 3,4-diphenyl-1,2,5-oxadiazoles (3,4-diphenylfurazans) analogs, were synthesized for in vitro evaluation as hybrid cyclooxygenase (COX) inhibitor/nitric oxide donor agents. Reaction of 1-[4-(methylsulfonyl)phenyl]-2 phenylethene with an aqueous sodium nitrite solution in acetic acid afforded a mixture (3:1 ratio) of the inseparable 4-[4-(methylsulfonyl)phenyl]-3-phenyl 1,2,5-oxadiazole-2-oxide (13a) and 3-[4-(methylsulfonyl)phenyl]-4-phenyl-1,2,5 oxadiazole-2-oxide (13b) regioisomers. A group of related regioisomers possessing either a p-aminosulfonylphenyl (16) or a p-azidosulfonylphenyl (17), moiety were obtained by chlorosulfonation of the unsubstituted 3,4-diphenylfuroxan (10) and subsequent reaction with either ammonium hydroxide or sodium azide, respectively. The methanesulfonyl regioisomers 13a,b [COX-1 IC50=11.6 microM; COX-2 IC50=0.12 microM; COX-2 selectivity index (SI)=97] and aminosulfonyl regioisomers 16 (COX-1 IC50=9.8 microM; COX-2 IC50=0.78 microM; COX-2 SI=12), like the reference drug celecoxib (COX-1 IC50=33.1 microM; COX-2 IC50=0.07 microM; COX-2 SI=472), were potent in vitro COX-2 inhibitors with a good COX-2 selectivity index. Release of nitric oxide (NO) from the 3,4-diphenylfuroxan compounds (10, 13a,b, 16, 17) was thiol-dependent since the % NO released was higher upon incubation in the presence of l-cysteine (0.57-3.18%) compared to that in phosphate buffer solution at pH7.4 (0.06-0.15%). Molecular modeling (docking) studies show that the methanesulfonyl (MeSO2) COX-2 pharmacophore present in regioisomers 13a,b is positioned in the vicinity of the COX-2 secondary pocket. The in vitro NO release data, COX-1/COX-2 inhibition and COX-2 SI structure-activity relationships acquired, and molecular modeling docking studies suggest that the 1,2,5 oxadiazole-2-oxide (furoxan) ring possesses beneficial features that should be present in a suitable central ring template (bioisostere) pertinent to the design novel hybrid COX-2 inhibitor/nitric oxide donor agents with a low ulcerogenicity profile that may be free from adverse cardiovascular effects. PMID- 15781388 TI - 3D-QSAR CoMFA studies on trypsin-like serine protease inhibitors: a comparative selectivity analysis. AB - A series of indole/benzoimidazole-5-carboxamidines have been reported to inhibit various trypsin-like serine proteases viz. uPA, tPA, factor Xa, thrombin, plasmin, and trypsin, which are involved in various types of pathophysiological conditions such as cancer progression, thrombosis etc. Inhibition of these protease enzymes may serve as therapeutic agents in various types of cancer as well serve as anticoagulant or antithrombotic agents. The dual inhibitory action may result in poor clinical candidates. 3D-QSAR models were generated for indole/benzoimidazole-5-carboxamidines using the CoMFA technique to study their selectivity trends toward various trypsin-like serine proteases. Molecular superimposition was carried out on the template structure using atom-based RMS fit method. The CoMFA models were established from the training set of 25-29 molecules and validated by predicting the activities of seven-eight test set molecules. The CoMFA models generated using steric and electrostatic fields for tPA, fXa, thrombin, plasmin, and trypsin inhibition exhibited better statistical significance than the CoMFA models generated using ClogP as an additional descriptor. Thus, the validated CoMFA models with steric and electrostatic fields were used to generate 3D contour maps, which may provide possible modification of molecules for better selectivity/activity. The present 3D-QSAR studies emphasize the selectivity trends of indole/benzoimidazole-5-carboxamidines, which may be obliging in designing novel selective serine protease inhibitors of therapeutic interest. PMID- 15781387 TI - Systematic synthesis of galloyl-substituted procyanidin B1 and B2, and their ability of DPPH radical scavenging activity and inhibitory activity of DNA polymerases. AB - Six galloyl-substituted procyanidin B1 and B2, 3-O-gallate, 3''-O-gallate, and 3,3''-di-O-gallate, were systematically synthesized with the condensation method using TMSOTf as a catalyst. Their ability of DPPH radical scavenging activity and DNA polymerase inhibitory activity were also investigated. The results indicated that the galloyl group of these compounds is very important for both activities. 3,3''-Di-O-gallate dimers acted as strong inhibitor against DNA polymerase alpha and beta, whereas the desgalloyl and monogalloyl compounds did not exhibit any appreciable inhibitory activity against the DNA polymerase beta. PMID- 15781389 TI - Design, synthesis, and anti-tumor activity of (2-O-alkyloxime-3-phenyl)-propionyl 1-O-acetylbritannilactone esters. AB - The extracts of Inula britannica have anti-inflammatory, anti-bacterial, anti hepatitic, and anti-tumor activities. Various sesquiterpene lactones with cytotoxic properties including 1-O-acetylbritannilactone (1) have been isolated from this Chinese medicinal plant. Eight derivatives of 1-O acetylbritannilactone, (2-O-alkyloxime-3-phenyl)-propionyl-1-O acetylbritannilactone esters were designed and synthesized. Four of these compounds were tested to show inhibitory activity on the growth of human leukemia HL-60 and cancer Bel-7402 cell lines. PMID- 15781390 TI - The novel GABA adamantane derivative (AdGABA): design, synthesis, and activity relationship with gabapentin. AB - A facile preparation of 2-aminomethyl-2-tricyclo[3.3.1.1(1,7)]decaneacetic acid hydrochloride 5 (AdGABA) is described. The synthesis of AdGABA involves the hydrogenation of 2-cyano-2-tricyclo[3.3.1.1(1,7)]decaneacetic acid 11, which was synthesized by two different synthetic routes. AdGABA was found to antagonize the pentylenetetrazole (PTZ) and semicarbazide (SCZ) induced tonic convulsions and exhibits analgesic activity in the hot plate test on mice. Although its mechanism of action is quite similar to that proposed previously for gabapentin (interaction with the alpha2delta subunit of the voltage gated Ca2+ channels), further studies were undertaken in order to clarify the precise mechanism of the anticonvulsant and analgesic effects of AdGABA on a molecular level. PMID- 15781391 TI - Design, synthesis, and biological evaluation of folic acid targeted tetraphenylporphyrin as novel photosensitizers for selective photodynamic therapy. AB - Photodynamic therapy (PDT) is a cancer treatment involving systemic administration of a tumor-localizing photosensitizer; this, when activated by the appropriate light wavelength, interacts with molecular oxygen to form a toxic, short-lived species known as singlet oxygen, which is thought to mediate cellular death. Targeted PDT offers the opportunity of enhancing photodynamic efficiency by directly targeting diseased cells and tissues. Two new conjugates of three components, folic acid/hexane-1,6-diamine/4-carboxyphenylporphyrine 1 and folic acid/2,2'-(ethylenedioxy)-bis-ethylamine/4-carboxyphenylporphyrine 2 were synthesized. The conjugates were characterized by 1H NMR, MALDI, UV-visible spectroscopy, and fluorescence quantum yield. The targeted delivery of these photoactive compounds to KB nasopharyngeal cell line, which is one of the numerous tumor cell types that overexpress folate receptors was studied. It was found that after 24 h incubation, conjugates 1 and 2 cellular uptake was on average 7-fold higher than tetraphenylporphyrin (TPP) used as reference and that 1 and 2 cellular uptake kinetics increased steadily over the 24 h period, suggesting an active transport via receptor-mediated endocytosis. In corresponding results, conjugates 1 and 2 accumulation displayed a reduction of 70% in the presence of a competitive concentration of folic acid. Survival measurements demonstrated that KB cells were significantly more sensitive to conjugated porphyrins-mediated PDT. Under the same experimental conditions and the same photosensitizer concentration, TPP displayed no photocytotoxicity while conjugates 1 and 2 showed photodynamic activity with light dose values yielding 50% growth inhibition of 22.6 and 6.7 J/cm2, respectively. PMID- 15781392 TI - Structure-activity relationships and molecular modelling of 5-arylidene-2,4 thiazolidinediones active as aldose reductase inhibitors. AB - The structure-activity relationships (SARs) of 5-arylidene-2,4-thiazolidinediones active as aldose reductase inhibitors (ARIs) were extended by varying the substitution pattern on the 5-arylidene moiety and on N-3. In particular, the introduction of an additional aromatic ring or an H-bond donor group on the 5 benzylidene ring enhanced ALR2 inhibitory potency. Moreover, the presence of a carboxylic anionic chain on N-3 was shown to be an important, although not essential, structural requisite to produce high levels of ALR2 inhibition. The length of this carboxylic chain was critical and acetic acids 4 were the most effective inhibitors among the tested derivatives. Molecular docking simulations into the ALR2 active site accorded with the in vitro inhibition data. They allowed the rationalization of the observed SARs and provided a pharmacophoric model for this class of ARIs. PMID- 15781393 TI - Targeting the gatekeeper residue in phosphoinositide 3-kinases. AB - A single residue in the ATP binding pocket of protein kinases-termed the gatekeeper-has been shown to control sensitivity to a wide range of small molecule inhibitors (Chem. Biol.2004, 11, 691; Chem. Biol.1999, 6, 671). Kinases that possess a small side chain at this position (Thr, Ala, or Gly) are readily targeted by structurally diverse classes of inhibitors, whereas kinases that possess a larger residue at this position are broadly resistant. Recently, lipid kinases of the phosphoinositide 3-kinase (PI3-K) family have become the focus of intense research interest as potential drug targets (Chem. Biol.2003, 10, 207; Curr. Opin. Pharmacol.2003, 3, 426). In this study, we identify the residue that corresponds structurally to the gatekeeper in PI3-Ks, and explore its importance in controlling enzyme activity and small molecule sensitivity. Isoleucine 848 of p110alpha was mutated to alanine and glycine, but the mutated kinase was found to have severely impaired enzymatic activity. A structural bioinformatic comparison of this kinase with its yeast orthologs identified second site mutations that rescued the enzymatic activity of the I848A kinase. To probe the dimensions of the gatekeeper pocket, a focused panel of analogs of the PI3-K inhibitor LY294002 was synthesized and its activity against gatekeeper mutated and wild-type p110alpha was assessed. PMID- 15781394 TI - Novel non-steroidal/non-anilide type androgen antagonists: discovery of 4 substituted pyrrole-2-carboxamides as a new scaffold for androgen receptor ligands. AB - We designed and synthesized novel pyrrole-2-carboxamide derivatives as androgen antagonists. Compounds 10 and 13 bearing benzylamine or aniline at the 4-position of the pyrrole ring showed moderate androgen antagonistic activity, and inhibited the androgen-dependent growth of Shionogi carcinoma cells (SC-3). Study of the structure-activity relationships of compound 13 led to a potent androgen antagonist 36, which has higher affinity than flutamide (4) for androgen nuclear receptor (AR). Thus, pyrrole-2-carboxamide is a new scaffold for developing AR antagonists. PMID- 15781395 TI - Synthesis and biological evaluation of penam sulfones as inhibitors of beta lactamases. AB - The chemical synthesis of a series of new penam sulfone derivatives bearing a 2beta-substituted-oxyimino and -hydrazone substituents, their beta-lactamase inhibitory properties against selected enzymes representing class A and C beta lactamases are reported. The oxime containing penam sulfones strongly inhibited the Escherichia coli TEM-1 and Klebsiella pneumoniae cefotaximase (CTX-1) enzymes, but moderately inhibited the Pseudomonas aeruginosa 46012 cephalosporinase; while the 2beta-substituted-hydrazone derivatives were generally less active against these enzymes. Furthermore, most of the inhibitors enhanced the antibacterial activities of piperacillin (PIP) and ceftazidime (CAZ) particularly against TEM-1 and CTX-1 producing bacterial strains. PMID- 15781397 TI - Design, synthesis and biological evaluation of novel riccardiphenol analogs. AB - A novel, facile, high yield, and less cumbersome synthesis of riccardiphenol analogs is described. The synthesized compounds were characterized and assessed for its in vitro activity in a panel of human cancer cell lines of differing origin: HuCCT-1, BxPC3, Panc-1, Mia-Paca, A431, Hep2, and HN006. HuCCT-1 was derived from an intrahepatic cholangiocarcinoma; BxPC3, Mia-Paca, and Panc-1 were derived from pancreatic cancers; A431 was derived from a vulvar epithelial carcinoma; and Hep2 and HN006 were derived from squamous cell carcinomas of the head and neck. The cytotoxicity of a newly developed riccardiphenol analog against human cancer cell lines was assessed. The cancer cells exhibited varying sensitivities to the compound, with IC50 values from 30 to 50 microM. This susceptibility was particularly interesting in the case of lines such as Hep2 and BxPC3 that are resistant to classic cytotoxic drugs as well as some targeted agents. These results demonstrate that the novel riccardiphenol analog has effective action against human-derived cancer cell in vitro. PMID- 15781396 TI - Structure based design of 4-(3-aminomethylphenyl)piperidinyl-1-amides: novel, potent, selective, and orally bioavailable inhibitors of betaII tryptase. AB - Tryptase is a serine protease found almost exclusively in mast cells. It has trypsin-like specificity, favoring cleavage of substrates with an arginine (or lysine) at the P1 position, and has optimal catalytic activity at neutral pH. Current evidence suggests tryptase beta is the most important form released during mast cell activation in allergic diseases. It is shown to have numerous pro-inflammatory cellular activities in vitro, and in animal models tryptase provokes broncho-constriction and induces a cellular inflammatory infiltrate characteristic of human asthma. Screening of in-house inhibitors of factor Xa (a closely related serine protease) identified beta-amidoester benzamidines as potent inhibitors of recombinant human betaII tryptase. X-ray structure driven template modification and exchange of the benzamidine to optimize potency and pharmacokinetic properties gave selective, potent and orally bioavailable 4-(3 aminomethyl phenyl)piperidinyl-1-amides. PMID- 15781398 TI - Atom, atom-type, and total nonstochastic and stochastic quadratic fingerprints: a promising approach for modeling of antibacterial activity. AB - The TOpological MOlecular COMputer Design (TOMOCOMD-CARDD) approach has been introduced for the classification and design of antimicrobial agents using computer-aided molecular design. For this propose, atom, atom-type, and total quadratic indices have been generalized to codify chemical structure information. In this sense, stochastic quadratic indices have been introduced for the description of the molecular structure. These stochastic fingerprints are based on a simple model for the intramolecular movement of all valence-bond electrons. In this work, a complete data set containing 1006 antimicrobial agents is collected and presented. Two structure-based antibacterial activity classification models have been generated. The models (including nonstochastic and stochastic indices) classify correctly more than 90% of 1525 compounds in training sets. These models permit the correct classification of 92.28% and 89.31% of 505 compounds in an external test sets. The TOMOCOMD-CARDD approach, also, satisfactorily compares with respect to nine of the most useful models for antimicrobial selection reported to date. Finally, a virtual screening of 87 new compounds reported in the antiinfective field with antibacterial activities is developed showing the ability of the TOMOCOMD-CARDD models to identify new leads as antibacterial. PMID- 15781399 TI - Mild and efficient oxidation of alcohols with sodium periodate catalyzed by polystyrene-bound Mn(III)porphyrin. AB - Mild and efficient oxidation of primary and secondary alcohols with sodium periodate catalyzed by Mn(TPyP), [manganese(III)tetra(4-pyridyl)porphyrin], supported on chloromethylated polystyrene, [Mn(TPyP)-CMP], at room temperature were reported. This catalyst can be reused consecutively four times in the oxidation of 4-chlorobenzyl alcohol in 97% yield without significant loss of its activity. PMID- 15781400 TI - Synthesis and human NKT cell stimulating properties of 3-O-sulfo-alpha/beta galactosylceramides. AB - Two novel hybrid molecules 3-O-sulfo-alpha/beta-galactosylceramide 3 and 4, which are derived from an immunostimulatory agent alpha-GalCer 1 and self-glycolipid ligand sulfatide 2, were designed and synthesized. Compound 3 was shown to efficiently stimulate human NKT cells to secret IL-4 and IFN-gamma, with activities similar to 1, suggesting that modification of the 3''-OH position of the galactose moiety with sulfate has no significant effect on NKT cell stimulation. As a comparison, the beta-isomer 4 has no affinity to NKT cells, which demonstrates that the alpha-glycosidic bond of galactosylceramide is crucial to the NKT cells activation. PMID- 15781401 TI - Synthesis of 9-alkyl and 9-heteroalkyl substituted 2-amino-6-guanidinopurines and their influence on the NO-production in macrophages. AB - 9-Alkyl and 9-heteroalkyl substituted derivatives of the 2-amino-6 guanidinopurine were synthesized by alkylation of 2-amino-6-chloropurine and subsequent guanidinolysis. The activity of the thus prepared compounds on murine macrophages was examined. Compounds 4a, 4b, and 4d inhibit the LPS+IFN-gamma induced NO production in murine macrophages while compound 4h stimulates this production. PMID- 15781402 TI - (R)-Goniothalamin: total syntheses and cytotoxic activity against cancer cell lines. AB - The total syntheses of (R)-goniothalamin (1), a styryl lactone isolated from several Goniothalamus species, via catalytic asymmetric allylation of alpha benzyloxyacetaldehyde (2), followed by ring-closing metathesis and Wittig olefination and via catalytic asymmetric allylation of trans-cinnamaldehyde (12), followed by ring-closing metathesis are reported. The antiproliferative activities of (R)-1 and its Z-isomer 10 as well as of the synthetic dihydropyranone intermediates 7 and 8 against eight different cancer cell lines are also described. PMID- 15781403 TI - 1,2,4-Triazino-[5,6b]indole derivatives: effects of the trifluoromethyl group on in vitro antimalarial activity. AB - In an attempt to search for new and alternative antimalarial agents, a series of unsubstituted and 6-trifluoromethyl-1,2,4-triazino[5,6b]indole and 5H-1,2,4 triazolo[1',5',2,3]-1,2,4-triazino[5,6b]indole derivatives were synthesized and their chemical structures confirmed by 1H NMR and 13C NMR, elemental, IR and mass spectrophotometric analyses. The in vitro antimalarial activities of these compounds were evaluated against the chloroquine-sensitive (D10) and the chloroquine-resistant (RSA11) strains of Plasmodium falciparum. The 1,2,4 triazino[5,6b]indole derivatives (4, 6 and 8) with a trifluoromethyl group at position 6 exhibit increased in vitro activity when compared to the unsubstituted analogues, which are all devoid of activity. The presence of the trifluoromethyl group in the 5H-1,2,4-triazolo[1',5',2,3]-1,2,4-triazino[5,6b]indole ring system leads to compounds with diminished antimalarial activity when compared to the corresponding unsubstituted analogues. The compounds associate with ferriprotoporphyrin IX and interact with DNA to more or less the same extent. PMID- 15781404 TI - Novel fluoropeptidomimetics: synthesis, stability studies and protease inhibition. AB - Designer fluoropeptidomimetics as protease inhibitors are revealed. The key peptidomimetic region in the inhibitors contains a '-CHF-S-' moiety and is designed to mimic the tetrahedral oxyanion species during the hydrolysis of a peptide bond. Designed fluoropeptidomimetics in aqueous methanol slowly (in several hours to days) yielded the corresponding methyl ether and/or the oxazole derivatives after cyclization. Alkyl substitutions at the C-2 position exhibited enhanced aqueous stability. Nature of '-CHF-S-' moiety and the stabilities of various fluoropeptidomimetics in aqueous solution are disclosed in detail. Fluoropeptidomimetics containing bulky substitutions at P1 such as compounds 15 and 16 exhibited time-dependent loss of activities against chymotrypsin, upto [corrected] 67% and 79% with a Ki of 63 and 120 microM, respectively. Fluoropeptidomimetics are a novel class of protease inhibitors and the next generation of fluoropeptidomimetics should incorporate enhanced stability. PMID- 15781405 TI - Novel A-ring analogs of the hormone 1alpha,25-dihydroxyvitamin D3: synthesis and preliminary biological evaluation. AB - Prepared from a commercial prostaglandin building block, novel vitamin D3 analogs with a contracted five-membered A-ring were designed and synthesized to mimic the A-ring diol structure of the natural hormone 1alpha,25-dihydroxyvitamin D3. Prepared from commercial 1,4-cyclohexanedione, a structurally simplified analog was designed and synthesized in which a suitably oriented primary allylic hydroxyl group at the C-2 position might be a surrogate for the biologically important 1alpha-OH in the natural hormone. PMID- 15781406 TI - QSAR by LFER model of cytotoxicity data of anti-HIV 5-phenyl-1-phenylamino-1H imidazole derivatives using principal component factor analysis and genetic function approximation. AB - Cytotoxicity data of anti-HIV 5-phenyl-1-phenylamino-1H-imidazole derivatives were subjected to quantitative structure-activity relationship (QSAR) study using linear free energy related (LFER) model of Hansch using electronic (Hammett sigma), hydrophobicity (pi) and steric (molar refractivity and STERIMOL L, B1, B2, B3 and B4) parameters of phenyl ring substituents of the compounds, along with appropriate indicator variables. Principal component factor analysis (FA) was used as the data-preprocessing step to identify the important predictor variables contributing to the response variable and to avoid collinearities among them. The generated multiple linear regression (MLR) equations were statistically validated using leave-one-out technique. Genetic function approximation (GFA) was also used on the same data set to develop QSAR equations, which produced the same best equation as obtained with FA-MLR. The final equation is of acceptable statistical quality (explained variance 80.2%) and predictive potential (leave one-out predicted variance 74%). The analysis explores the structural and physicochemical contributions of the compounds for cytotoxicity. A thiol substituent at 2 position of the imidazole nucleus decreases cytotoxicity when compared to the corresponding unsubstituted congener. Presence of hydrogen bond donor group at meta position of the phenyl ring present at 5 position of the imidazole nucleus also reduces cytotoxicity. Additionally, absence of any substituent at 2 and 3 positions of the phenyl ring of 1-phenylamino fragment reduces the cytotoxicity. The negative coefficient of sigmap indicates that presence of electron-withdrawing substituents at the para position of the phenyl ring of the 1-phenylamino fragment is not favourable for the cytotoxicity. Again, lipophilicity of meta substituents of the 5-phenyl ring increases cytotoxicity. The coefficients of molar refractivity (MRm) and STERIMOL parameters for meta substituents (Lm, B1m and B4m) of the phenyl ring of 1-phenylamino fragment indicate that the length, width and overall size of meta substituents are conducive factors for the cytotoxicity. PMID- 15781407 TI - Studies on the cytochrome P450 catalyzed oxidation of 13C labeled 1-cyclopropyl-4 phenyl-1,2,3,6-tetrahydropyridine by 13C NMR. AB - A recent study from Hanzlik's laboratory (J. Am. Chem. Soc. 2002, 124, 8268) has provided compelling evidence of a hydrogen atom transfer pathway for the cytochrome P450-catalyzed oxidative N-decyclopropylation of N-cyclopropyl-N methylaniline. In the present paper, we report an analogous pathway for the oxidative decyclopropylation of a 13C-labeled 1-cyclopropyl-4-phenyl-1,2,3,6 tetrahydropyridinyl substrate. Three 13C-enriched metabolites were characterized: (1) a diastereomeric pair of N-cyclopropyl-N-oxides; (2) the N cyclopropylpyridinium species; and (3) cyclopropanone hydrate. These results extend the hydrogen atom transfer pathway to include aliphatic tertiary amine substrates. Consideration of all of the available evidence, however, leads us to conclude that the cytochrome P450-catalyzed alpha-carbon oxidations of cyclopropylamines may proceed via both the single electron and hydrogen atom transfer pathways. PMID- 15781408 TI - Investigations of linker structure on the potency of a series of bidentate protein tyrosine phosphatase inhibitors. AB - Protein tyrosine phosphatases (PTPases) and protein tyrosine kinase (PTKases) regulate the phosphorylation and dephosphorylation of tyrosine residues in proteins, events that are essential for a variety of cellular functions. PTPases such as PTP1B and the Yersinia PTPase play an important role in diseases including type II diabetes and bubonic plague. A library of 67 bidentate PTPase inhibitors that are based on the alpha-ketocarboxylic acid motif has been synthesized using parallel solution-phase methods. Two aryl alpha-ketocarboxylic acids were tethered to a variety of different diamine linkers through amide bonds. The compounds were assayed in crude form against the Yersinia PTPase, PTP1B, and TCPTP. Six compounds were selected for further evaluation, in purified form, against the Yersinia PTPase, PTP1B, TCPTP, LAR, and CD45. These compounds had IC50 values in the low micromolar range against the Yersinia PTPase, PTP1B, and TCPTP, showed good selectivity for PTP1B over LAR, and modest selectivity over CD45. The correlation between linker structure and inhibitor activity shows that aromatic groups in the linker can play an important role in determining binding affinity in this class of inhibitors. PMID- 15781409 TI - [11C]Glycylsarcosine: synthesis and in vivo evaluation as a PET tracer of PepT2 transporter function in kidney of PepT2 null and wild-type mice. AB - [11C]Glycylsarcosine (Gly-Sar) was synthesized as a potential radiotracer to investigate the localization and in vivo function of the peptide transporter PepT2 in mouse kidney. Its C-11 labeled diketopiperazine derivative, [11C]cyclo(Gly-Sar) [1-methylpiperazine-2,5-dione], was also evaluated as a potential tracer. [11C]Gly-Sar exhibited rapid initial uptake into kidneys with slow clearance from the medulla, consistent with uptake and retention of the radiotracer through the actions of PepT2. In contrast, the corresponding cyclized dipeptide [11C]cyclo(Gly-Sar) showed rapid clearance and accumulation only in the renal pelvis region. Involvement of PepT2 in reabsorption and delayed clearance of [11C]Gly-Sar was confirmed using the PepT2 knockout mouse, where rapid renal elimination of [11C]Gly-Sar and the absence of radioactivity in medulla were observed. This study demonstrates using in vivo imaging technique that PepT2 is primarily responsible for renal tubular active reabsorption of Gly-Sar, and provides a new tool for studying tubular peptide reabsorption and clearance. PMID- 15781410 TI - Protein linear indices of the 'macromolecular pseudograph alpha-carbon atom adjacency matrix' in bioinformatics. Part 1: prediction of protein stability effects of a complete set of alanine substitutions in Arc repressor. AB - A novel approach to bio-macromolecular design from a linear algebra point of view is introduced. A protein's total (whole protein) and local (one or more amino acid) linear indices are a new set of bio-macromolecular descriptors of relevance to protein QSAR/QSPR studies. These amino-acid level biochemical descriptors are based on the calculation of linear maps on Rn[f k(xmi):Rn-->Rn] in canonical basis. These bio-macromolecular indices are calculated from the kth power of the macromolecular pseudograph alpha-carbon atom adjacency matrix. Total linear indices are linear functional on Rn. That is, the kth total linear indices are linear maps from Rn to the scalar R[f k(xm):Rn-->R]. Thus, the kth total linear indices are calculated by summing the amino-acid linear indices of all amino acids in the protein molecule. A study of the protein stability effects for a complete set of alanine substitutions in the Arc repressor illustrates this approach. A quantitative model that discriminates near wild-type stability alanine mutants from the reduced-stability ones in a training series was obtained. This model permitted the correct classification of 97.56% (40/41) and 91.67% (11/12) of proteins in the training and test set, respectively. It shows a high Matthews correlation coefficient (MCC=0.952) for the training set and an MCC=0.837 for the external prediction set. Additionally, canonical regression analysis corroborated the statistical quality of the classification model (Rcanc=0.824). This analysis was also used to compute biological stability canonical scores for each Arc alanine mutant. On the other hand, the linear piecewise regression model compared favorably with respect to the linear regression one on predicting the melting temperature (tm) of the Arc alanine mutants. The linear model explains almost 81% of the variance of the experimental tm (R=0.90 and s=4.29) and the LOO press statistics evidenced its predictive ability (q2=0.72 and scv=4.79). Moreover, the TOMOCOMD-CAMPS method produced a linear piecewise regression (R=0.97) between protein backbone descriptors and tm values for alanine mutants of the Arc repressor. A break-point value of 51.87 degrees C characterized two mutant clusters and coincided perfectly with the experimental scale. For this reason, we can use the linear discriminant analysis and piecewise models in combination to classify and predict the stability of the mutant Arc homodimers. These models also permitted the interpretation of the driving forces of such folding process, indicating that topologic/topographic protein backbone interactions control the stability profile of wild-type Arc and its alanine mutants. PMID- 15781411 TI - In silico ADME modelling: prediction models for blood-brain barrier permeation using a systematic variable selection method. AB - Quantitative Structure-Property Relationship models (QSPR) based on in vivo blood brain permeation data (logBB) of 88 diverse compounds, 324 descriptors and a systematic variable selection method, namely 'Variable Selection and Modeling method based on the prediction (VSMP)', are reported. Of all the models developed using VSMP, the best three-descriptors model is based on Atomic type E-state index (SsssN), AlogP98 and Van der Waal's surface area (r=0.8425, q=0.8239, F=68.49 and SE=0.4165); the best four-descriptors model is based on Kappa shape index of order 1, Atomic type E-state index (SsssN), Atomic level based AI topological descriptor (AIssssC) and AlogP98 (r=0.8638, q=0.8472, F=60.982 and SE=0.3919). The performance of the models on three test sets taken from the literature is illustrated and compared with the results from other reported computational approaches. Test set III constitutes 91 compounds from the literature with known qualitative BBB indication and is used for virtual screening studies. The success rate of the reported models is 82% in the case of BBB+ compounds and a similar success rate is observed with BBB- compounds. Finally, as the models reported herein are based on computed properties, they appear as a valuable tool in virtual screening, where selection and prioritization of candidates is required. PMID- 15781412 TI - Parallel solid-phase synthesis and characterization of new sulfonamide and carboxamide proline derivatives as potential CNS agents. AB - A solid-phase synthesis of the 64-member library of novel sulfonamide and carboxamide proline derivatives, focused on the 5-HT7 receptor antagonist SB 258741, was described. The final compounds were obtained in good yields and high purity upon cleavage from SynPhase Lanterns, functionalized by a BAL linker. The library representatives were screened for 5-HT7, 5-HT1A and D2 receptors to explore the impact of a tertiary amine moiety, the length of an alkylene spacer and the aryl fragment on the receptor affinity. The preliminary biological results provided data for further investigation aimed at a search for 5-HT7 receptor agents, and permitted the identification of several compounds with significant 5-HT1A receptor affinity. PMID- 15781413 TI - Synthesis, properties, and photodynamic inactivation of Escherichia coli using a cationic and a noncharged Zn(II) pyridyloxyphthalocyanine derivatives. AB - The photodynamic effect of a cationic Zn(II) N-methylpyridyloxyphthalocyanine (ZnPc 2) and a noncharged Zn(II) pyridyloxyphthalocyanine (ZnPc 1) has been compared in both homogeneous media bearing photooxidizable substrates and in vitro using a typical Gram-negative bacterium Escherichia coli. Absorption and fluorescence spectroscopic studies were analyzed in different media. Fluorescence quantum yields (varphiF) of 0.23 for ZnPc 1 and 0.22 for ZnPc 2 were calculated in N,N-dimethylformamide (DMF). The singlet molecular oxygen, O2(1Deltag), production was evaluated using 9,10-dimethylanthracene (DMA) in DMF yielding values of PhiDelta=0.56 for ZnPc 1 and 0.59 for ZnPc 2. A faster decomposition of L-tryptophan (Trp), which was used as biological substrate model, was obtained using ZnPc 2 as a sensitizer with respect to ZnPc 1. In biological medium, the E. coli cultures were treated with 10 microM of sensitizer for different times at 37 degrees C in the dark. Both ZnPcs 1 and 2 are rapidly bound to E. coli cells in 5 min and the amount of cell-bound sensitizer is not appreciably changed incubating the cultures for longer times. The recovered ZnPc 2 after one washing step is approximately 3 times higher than 1, reaching a value of approximately 3 nmol/10(6) cells. After irradiation with visible light, a higher photoinactivation of cells was found for ZnPc 2. Thus, a approximately 4.5 log (99.997%) decrease of cell survival was obtained after 30 min of irradiation. On the other hand, a very low photodamage was found for cells treated with ZnPc 1 (approximately 0.5 log). Also, these results were established by stopping of growth curves for E. coli. In the structure of ZnPc 2, the cationic centers are isolated from the phthalocyanine ring by an ether bridge, which also provides a higher mobility of the charges facilitating the interaction with the outer membrane of the Gram-negative bacteria. These studies show that cationic ZnPc 2 is an efficient phototherapeutic agent with potential applications in photodynamic inactivation of bacteria. PMID- 15781414 TI - Chiral cyclopalladated complexes derived from N,N-dimethyl-1-phenethylamine with bridging bis(diphenylphosphine)ferrocene ligand as inhibitors of the cathepsin B activity and as antitumoral agents. AB - Chiral cyclopalladated complexes derived from N,N-dimethyl-1-phenethylamine and the coordinating ligand 1,1'-bis(diphenylphosphine)ferrocene were synthesized and studied as Cathepsin B inhibitors and antitumoral agents against solid tumors. Our results revealed that the palladium compound [Pd2(C2,N-S(-)dmpa)2(mu dppf)Cl2] (2) was able to inhibit Cathepsin B activity in a reversible fashion. This palladacycle compound binds to free cathepsin B (E) as well as to the enzyme substrate complex (ES) with dissociation constants of KH=12+/-1 microM and alphaKH=2.4+/-0.3 microM, respectively. The application of this complex, in Walker tumor-bearing rats, resulted in 90% inhibition of the tumor growth. Subcutaneous inoculations of 10(6) tumoral cells produced solid tumors with a mass of 4.0+/-1.0 g in 12 days Walker tumor-bearing rats. However, when these animals were treated with one dose of the palladacycle compound (2.0 mg/kg), the tumoral mass was reduced to 0.3+/-0.1 g. On the other hand, the same complex (2) did not afford any protection to mice bearing the non-metastatic Ehrlich Ascites tumor treated with doses of 0.5, 5.0, and 30 mg/kg for a period of four, three and one day, respectively, beginning 72 h after tumor inoculation. Toxicological studies using mice treated with one high dose of the complex (2) (100 mg/kg) did not show any alterations in red and white blood cell morphology 14 days after the drug administration. Similar results were obtained with hepatic, kidney, and spleen tissues. The results presented in this work introduce the title cyclopalladated complexes as promising antitumoral drugs with reduced toxicity in experimental studies. PMID- 15781415 TI - Isochromanone-based urotensin-II receptor agonists. AB - A series of analogues of the selective non-peptide urotensin II (UII) receptor agonist 3-(4-chlorophenyl)-3-(2-dimethylaminoethyl)-isochroman-1-one (AC-7954, 1) was synthesized and evaluated for UII agonist activity using a functional cell based assay. The introduction of a methyl group in the 4-position resulted in a complete loss of activity, whereas substituents in the aromatic rings were beneficial. Sterically demanding amino groups were also detrimental to the activity. Several potent agonists were identified, six compounds being equally or more potent than 1. The most potent compound in the series was the 6,7-dimethyl analogue of 1 (16, pEC50 6.87). The racemate of 16 was resolved into the pure enantiomers using preparative straight phase HPLC. It was shown that the potency resides in the (+)-enantiomer (pEC50 7.11). The synthesized compounds seem to be selective for the UII receptor as no activities were observed at the closely related SSTR3 and 5 receptors. PMID- 15781416 TI - Comparative study of flux redistribution of metabolic pathway in glutamate production by two coryneform bacteria. AB - In amino acid production by coryneform bacteria, study on relationship between change in enzyme activities and production of a target amino acid is important. In glutamate production, Kawahara et al. discovered that the effect of decrease in 2-oxoglutamate dehydrogenase complex (ODHC) on glutamate production is essential (Kawahara et al., Biosci. Biotechnol. Biochem. 61(7) (1997) 1109). Significant reduction of the ODHC activity was observed in the cells under the several glutamate-productive conditions in Corynebacterium glutamicum. Recent progress in metabolic engineering enables us to quantitatively compare the flux redistribution of the different strains after change in enzyme activity precisely. In this paper, relationship between flux redistribution and change in enzyme activities after biotin deletion and addition of detergent (Tween 40) was studied in two coryneform bacteria, C. glutamicum and a newly isolated strain, Corynebacterium efficiens (Fudou et al., Int. J. Syst. Evol. Microbiol. 52(Part 4) 1127), based on metabolic flux analysis (MFA). It was observed that in both species the specific activities of isocitrate dehydrogenase (ICDH) and glutamate dehydrogenase (GDH) did not significantly change throughout the fermentation, while that of the ODHC significantly decreased after biotin depletion and Tween 40 addition. Flux redistribution clearly occurred after the decrease in ODHC specific activity. The difference in glutamate production between C. glutamicum and C. efficiens was caused by the difference in the degree of decrease in ODHC specific activity. The difference in Michaelis-Menten constants or K(m) value between ICDH, GDH, and ODHC explained the mechanism of flux redistribution at the branch point of 2-oxoglutarate. It was found that the K(m) values of ICDH and ODHC were much lower than that of GDH for both strains. It was quantitatively proved that the ODHC plays the most important role in controlling flux distribution at the key branch point of 2-oxoglutarate in both coryneform bacteria. Flux redistribution mechanism was well simulated by a Michaelis-Menten based model with kinetic parameters. The knowledge of the mechanism of flux redistribution will contribute to improvement of glutamate production in coryneform bacteria. PMID- 15781417 TI - Adaptation for fast growth on glucose by differential expression of central carbon metabolism and gal regulon genes in an Escherichia coli strain lacking the phosphoenolpyruvate:carbohydrate phosphotransferase system. AB - Phosphoenolpyruvate (PEP) is a key intermediate of cellular metabolism and a precursor of commercially relevant products. In Escherichia coli 50% of the glucose-derived PEP is consumed by the PEP:carbohydrate phosphotransferase system (PTS) for glucose transport. PTS, encoded by the ptsHIcrr operon, was deleted from JM101 to generate strain PB11 (PTS-Glc-). PB12, a mutant derived from PB11, grows faster than the parental strain on glucose (PTS-Glc+ phenotype). This strain can redirect some of the PEP not utilized by PTS into the high yield synthesis of aromatic compounds from glucose. Here, we report a comparative transcription analysis among these strains of more than 100 genes involved in central carbon metabolism during growth on glucose. It was found that in the PTS- strains that have reduced glucose transport capacities, several genes encoding proteins with functions related to carbon transport and metabolism were upregulated. Therefore, it could be inferred that these strains synthesize autoinducers of these genes when sensing very low internal glucose concentrations, probably for scavenging purposes. This condition that is permanently present in the PTS- strains even when growing in high glucose concentrations allowed the simultaneous utilization of glucose and acetate as carbon sources. It was found that the gal operon is upregulated in these strains, as well as the aceBAK, poxB and acs genes among others. In PB12, glk, pgi, the TCA cycle and certain respiratory genes are also upregulated. A mutation in arcB in PB12 is apparently responsible for the upregulation of the TCA cycle and certain respiratory genes. PMID- 15781418 TI - Evolution of intrahepatic carbon, nitrogen, and energy metabolism in a D galactosamine-induced rat liver failure model. AB - A clearer picture of the hepatic metabolic pathways affected by fulminant hepatic failure (FHF) would help develop nutritional support and nonsurgical therapies for FHF. We characterized the evolution of hepatic metabolism in a rat model of FHF using an isolated perfused liver system together with a mass-balance model of intermediary metabolism. Principal component analysis (PCA) was used to identify potential new sensitive markers for FHF. To induce FHF, rats were given two D galactosamine injections under fasting conditions. Controls were fasted only. Livers were harvested 1, 4, 8, and 12 h later and perfused with Eagle minimal essential medium supplemented with amino acids and bovine serum albumin, and equilibrated with 95% O2/5% CO2. At the 1 h time point, lactate release increased concomitant with a decrease in gluconeogenesis, TCA cycle and mitochondrial electron transport fluxes. At 4 h, amino acid metabolism and urea cycle fluxes were significantly depressed. By 8 h, gluconeogenesis had switched to glycolysis. By 12 h, amino acid metabolism was broadly inhibited, and there was a net release of many amino acids. Mass-balance analysis shows that the main source of ATP production in the FHF liver gradually changed from mitochondrial oxidative phosphorylation to glycolysis. PCA suggests that a linear combination of glucose, lactate, and glutamine concentrations in arterial plasma is a sensitive marker for FHF. We conclude that D-galactosamine causes early mitochondrial dysfunction while glycolytic ATP synthesis remains functional. Markers that are indirectly linked to these pathways may be used to evaluate the progression of FHF. PMID- 15781419 TI - Effect of a single-gene knockout on the metabolic regulation in Escherichia coli for D-lactate production under microaerobic condition. AB - The effects of several single-gene knockout mutants (pykF, ppc, pflA, pta, and adhE mutants) on the metabolic flux distribution in Escherichia coli were investigated under microaerobic condition. The intracellular metabolite concentrations and enzyme activities were measured, and the metabolic flux distribution was computed to study the metabolic regulation in the cell. The pflA, pta and ppc mutants produced large amount of lactate when using glucose as a carbon source under microaerobic condition. Comparing the flux distribution and the enzyme activities in the mutants, it was shown that the lactate production was promoted by the inactivation of pyruvate formate lyase and the resulting overexpression of lactate dehydrogenase. The flux through Pta-Ack pathways and the ethanol production were limited by the available acetyl coenzyme A. It was shown that the glycolysis was activated in pykF mutant in microaerobic culture. The glycolytic flux was related with Pyk activity except for pykF mutant. The cell growth rate was shown to be affected by the flux through phosphoenolpyruvate carboxylase. The quantitative regulation analysis was made based on the deviation indexes. PMID- 15781420 TI - Metabolic engineering of aerobic succinate production systems in Escherichia coli to improve process productivity and achieve the maximum theoretical succinate yield. AB - The potential to produce succinate aerobically in Escherichia coli would offer great advantages over anaerobic fermentation in terms of faster biomass generation, carbon throughput, and product formation. Genetic manipulations were performed on two aerobic succinate production systems to increase their succinate yield and productivity. One of the aerobic succinate production systems developed earlier (Biotechnol, Bioeng., 2004, accepted) was constructed with five mutations (DeltasdhAB, Deltaicd, DeltaiclR, DeltapoxB, and Delta(ackA-pta)), which created a highly active glyoxylate cycle. In this study, a second production system was constructed with four of the five above mutations (DeltasdhAB, DeltaiclR, DeltapoxB, and Delta(ackA-pta)). This system has two routes in the aerobic central metabolism for succinate production. One is the glyoxylate cycle and the other is the oxidative branch of the TCA cycle. Inactivation of ptsG and overexpression of a mutant Sorghum pepc in these two production systems showed that the maximum theoretical succinate yield of 1.0 mol/mol glucose consumed could be achieved. Furthermore, the two-route production system with ptsG inactivation and pepc overexpression demonstrated substantially higher succinate productivity than the previous system, a level unsurpassed for aerobic succinate production. This optimized system showed remarkable potential for large-scale aerobic succinate production and process optimization. PMID- 15781421 TI - gNCA: a framework for determining transcription factor activity based on transcriptome: identifiability and numerical implementation. AB - Network Component Analysis (NCA) is a network structure-driven framework for deducing regulatory signal dynamics. In contrast to classical approaches such as principal component analysis or independent component analysis, NCA makes use of the connectivity structure from transcriptional regulatory networks to restrict the decomposition to a unique solution. However, the existing version of NCA cannot incorporate information beyond the network topology such as information obtained from regulatory gene knockouts that constrain the dynamics of regulatory signals. The ability of incorporating such information enables a more accurate and self-consistent analysis over different experiments and extends NCA to systems that may not satisfy the identifiability criteria of NCA. In this paper, we derive a generalized form of NCA, gNCA, which significantly expands the capability of transcription network analysis by incorporating regulatory signal constraints arising from genetic knockouts. The theoretical bases including criteria for uniqueness of solution and distinguishability between networks are derived. In addition, numerical techniques for robust decomposition are discussed. gNCA is then demonstrated using an Escherichia coli wild-type strain and an isogenic arcA deletion mutant during a carbon source transition. PMID- 15781422 TI - Determination of elasticities, concentration and flux control coefficients from transient metabolite data using linlog kinetics. AB - This paper presents a practical approach to estimate the kinetic parameters of a metabolic network from in vivo kinetics experiments. This method is based on the linlog kinetics format (Visser and Heijnen, 2003, Metab. Eng. 5(3), 164-176; Wu et al., 2004, Eur. J. Biochem. 271, 3348-3359), of which the kinetic parameters, called elasticities, are estimated by an iterative linear optimization followed by non-linear optimization, from transient metabolite concentration data which are directly obtainable from rapid pulse experiments. In this way, not only the parameters are estimated but also a full kinetic model, based on linlog kinetics, is developed. The obtained elasticities also allow immediate calculation of all control coefficients. As an in silico case study, the estimation of elasticities of a linear pathway is presented. The method is shown to be able to estimate the elasticities quite accurately and to be robust toward errors in the metabolite data originating from sampling and measurement inaccuracy. The method allows experimental redesign to get more accurate estimated parameters and accommodates various types of experimentally applied disturbances in the pathway: changes in independent metabolites, dependent metabolites or enzyme levels/activities. PMID- 15781423 TI - Decreased expression of both the alpha1- and alpha2-subunits of the Na-K-ATPase reduces maximal alveolar epithelial fluid clearance. AB - Impaired epithelial sodium channel function predisposes to delayed resorption of pulmonary edema and more severe experimental lung injury, whereas even a small fraction of the normal Na-K-ATPase activity is thought to be sufficient to maintain normal ion transport. However, direct proof is lacking. Therefore, we studied baseline and cAMP stimulated alveolar fluid clearance (AFC) in mice with a 50% decrease in lung protein expression of the alpha(1)- and/or alpha(2) subunit of the Na-K-ATPase. There was no difference in basal and stimulated AFC in alpha(1)(+/-) or alpha(2)(+/-) mice compared with wild-type littermates. Also, the compound heterozygous mice (alpha(1)(+/-)/alpha(2)(+/-)) had normal basal AFC. However, the combined alpha(1)(+/-)/alpha(2)(+/-) mice showed a significant decrease in cAMP-stimulated AFC compared with wild-type littermates (11.1 +/- 1.0 vs. 14.9 +/- 1.8%/30 min, P < 0.001). When exposed to 96 h of >95% hyperoxia, the decrease in stimulated AFC in the alpha(1)(+/-)/alpha(2)(+/-) mice was not associated with more lung edema compared with wild-type littermates (lung wet-to dry weight ratio 6.6 +/- 0.9 vs. 5.9 +/- 1.1, respectively; P = not significant). Thus a 50% decrease in protein expression of the alpha(1)- or alpha(2)-subunits of the Na-K-ATPase does not impair basal or stimulated AFC. However, a 50% protein reduction in both the alpha(1)- and alpha(2)-subunits of the Na-K-ATPase produces a submaximal stimulated AFC, suggesting a synergistic role for alpha(1)- and alpha(2)-subunits in cAMP-dependent alveolar epithelial fluid clearance. PMID- 15781424 TI - A combined yeast/bacteria two-hybrid system: development and evaluation. AB - Two-hybrid screening is a standard method used to identify and characterize protein-protein interactions and has become an integral component of many proteomic investigations. The two-hybrid system was initially developed using yeast as a host organism. However, bacterial two-hybrid systems have also become common laboratory tools and are preferred in some circumstances, although yeast and bacterial two-hybrid systems have never been directly compared. We describe here the development of a unified yeast and bacterial two-hybrid system in which a single bait expression plasmid is used in both organismal milieus. We use a series of leucine zipper fusion proteins of known affinities to compare interaction detection using both systems. Although both two-hybrid systems detected interactions within a comparable range of interaction affinities, each demonstrated unique advantages. The yeast system produced quantitative readout over a greater dynamic range than that observed with bacteria. However, the phenomenon of "autoactivation" by baits was less of a problem in the bacterial system than in the yeast. Both systems identified physiological interactors for a library screen with a cI-Ras test bait; however, non-identical interactors were obtained in yeast and bacterial screens. The ability to rapidly shift between yeast and bacterial systems provided by these new reagents should provide a marked advantage for two-hybrid investigations. In addition, the modified expression vectors we describe in this report should be useful for any application requiring facile expression of a protein of interest in both yeast and bacteria. PMID- 15781425 TI - A mathematical approach to molecular organization and proteolytic disintegration of bacterial inclusion bodies. AB - The in vivo proteolytic digestion of bacterial inclusion bodies (IBs) and the kinetic analysis of the resulting protein fragments is an interesting approach to investigate the molecular organization of these unconventional protein aggregates. In this work, we describe a set of mathematical instruments useful for such analysis and interpretation of observed data. These methods combine numerical estimation of digestion rate and approximation of its high-order derivatives, modelling of fragmentation events from a mixture of Poisson processes associated with differentiated protein species, differential equations techniques in order to estimate the mixture parameters, an iterative predictor corrector algorithm for describing the flow diagram along the cascade process, as well as least squares procedures with minimum variance estimates. The models are formulated and compared with data, and successively refined to better match experimental observations. By applying such procedures as well as newer improved algorithms of formerly developed equations, it has been possible to model, for two kinds of bacterially produced aggregation prone recombinant proteins, their cascade digestion process that has revealed intriguing features of the IB-forming polypeptides. PMID- 15781426 TI - A hybrid mathematical model of solid tumour invasion: the importance of cell adhesion. AB - In this paper we present a hybrid mathematical model of the invasion of healthy tissue by a solid tumour. In particular we consider early vascular growth, just after angiogenesis has occurred. We examine how the geometry of the growing tumour is affected by tumour cell heterogeneity caused by genetic mutations. As the tumour grows, mutations occur leading to a heterogeneous tumour cell population with some cells having a greater ability to migrate, proliferate or degrade the surrounding tissue. All of these cell properties are closely controlled by cell-cell and cell-matrix interactions and as such the physical geometry of the whole tumour will be dependent on these individual cell interactions. The hybrid model we develop focuses on four key variables implicated in the invasion process: tumour cells, host tissue (extracellular matrix), matrix-degradative enzymes and oxygen. The model is considered to be hybrid since the latter three variables are continuous (i.e. concentrations) and the tumour cells are discrete (i.e. individuals). With this hybrid model we examine how individual-based cell interactions (with one another and the matrix) can affect the tumour shape and discuss which of these interactions is perhaps most crucial in influencing the tumour's final structure. PMID- 15781427 TI - Influence of tumours on normal cells and optimal chemotherapy regimens: the case of several drugs and toxicity constraints. AB - Cancer chemotherapy with the application of several drugs is studied. The negative and inhibiting effect of the tumour on normal cells is taken into account. Under certain hypotheses, we determine the optimal regimen that minimizes the tumour burden at the end of a fixed period of therapy while maintaining several normal cell populations above prescribed levels. More precisely, it is demonstrated that the optimal drug administration corresponds to the strategy of intensive chemotherapy. PMID- 15781428 TI - Relation of N-terminal pro B-type natriuretic peptide to progression of aortic valve disease. AB - AIMS: Recently an elevation of B-type natriuretic peptide (BNP) and its N terminal fragment (NT-proBNP) in patients with aortic stenosis (AS) and aortic regurgitation (AR) has been described. The objective of this study was to evaluate the relation of NT-proBNP values to the progression of aortic valve disease. METHODS AND RESULTS: One hundred and sixty-eight patients were included. NT-proBNP was elevated in patients with AS (n=109) and AR (n=37) linked to disease severity. Values for NT-proBNP, pressure gradient, and left ventricular mass were identical in patients (n=22) after previous valve replacement and in those patients with mild AS. NT-proBNP levels decreased in 86 patients after valve replacement (2292+/-353 vs. 785+/-101 pg/ml; P<0.01) but increased in 82 patients who were treated conservatively (616+/-120 vs. 1155+/-432 pg/mL; P=0.029), related to the progression of disease. CONCLUSION: NT-proBNP is elevated in patients with aortic valve disease linked to disease severity and decreases after successful surgical therapy but increases in conservatively treated patients. These data underline the consistent relation of NT-proBNP to severity of aortic valve disease. Therefore, NT-proBNP should be considered as a biomarker for the monitoring of disease during follow-up, but further studies are warranted. PMID- 15781429 TI - Efficacy and safety of the coadministration of ezetimibe with fenofibrate in patients with mixed hyperlipidaemia. AB - AIMS: To examine the efficacy and safety of coadministered ezetimibe (EZE) with fenofibrate (FENO) in patients with mixed hyperlipidaemia. METHODS AND RESULTS: This was a multicentre, randomized, double-blind, placebo-controlled, parallel arm trial in patients with mixed hyperlipidaemia [LDL-cholesterol (LDL-C), 3.4 5.7 mmol/L (2.6-4.7 mmol/L for patients with type 2 diabetes); triglycerides (TG), 2.3-5.7 mmol/L] and no history of coronary heart disease (CHD), CHD equivalent disease (except for type 2 diabetes), or CHD risk score>20%. A total of 625 patients was randomized in a 1:3:3:3 ratio to one of four daily treatments for 12 weeks: placebo; EZE 10 mg; FENO 160 mg; FENO 160 mg plus EZE 10 mg (FENO+EZE). The primary endpoint compared the LDL-C lowering efficacy of FENO+EZE vs. FENO alone. LDL-C, non-HDL-cholesterol (non-HDL-C), and apolipoprotein B were significantly (P<0.001) reduced with FENO+EZE when compared with FENO or EZE alone. TG levels were significantly decreased and HDL-C was significantly increased with FENO+EZE and FENO treatments when compared with placebo (P<0.001). Coadministration therapy reduced LDL-C by 20.4%, non-HDL-C by 30.4%, TG by 44.0%, and increased HDL-C by 19.0%. At baseline, >70% of all patients exhibited the small, dense LDL pattern B profile. A greater proportion of patients on FENO+EZE and FENO alone treatments shifted from a more atherogenic LDL size pattern to a larger, more buoyant, and less atherogenic LDL size pattern at study endpoint than those on placebo or EZE. All three active therapies were well tolerated. CONCLUSION: Coadministration of EZE with FENO provided a complementary efficacy therapy that improves the atherogenic lipid profile of patients with mixed hyperlipidaemia. PMID- 15781430 TI - Should brain natriuretic peptides be measured in patients with aortic valve disease? PMID- 15781431 TI - Management of mixed hyperlipidaemia. PMID- 15781432 TI - Value of community-derived risk models for stratifying patients with non-ST elevation acute coronary syndromes. PMID- 15781433 TI - Dilated cardiomyopathy: more genes means more phenotypes. PMID- 15781434 TI - 'This is a walking test, not a talking test': the six minute walking test in congestive heart failure. PMID- 15781435 TI - Recommendations for the structure, organization, and operation of intensive cardiac care units. AB - Two major changes in patient characteristics and management occurred recently that demand distinctive alterations in the function of the intensive cardiac care unit (ICCU). These changes include the introduction of an early invasive strategy for the treatment of acute coronary syndromes, enabling early recuperation and shorter need for intensive care on the one hand, while the number of older and sicker patients requiring prolonged and more complex intensive care is steadily increasing. A task force of the European Society of Cardiology Working Group on Acute Cardiac Care was set to give a modern updated comprehensive recommendations concerning the structure, organization, and function of the modern ICCUs and intermediate cardiac units. These include the statement that specially trained cardiologists and cardiac nurses who can manage patients with acute cardiac conditions should staff the ICCUs. The optimum number of physicians, nurses, and other personal working in the unit is included. The document indicates the desired architecture and structure of the units and the intermediate cardiac unit and their relations to the other facilities in the hospital. Specific recommendations are also included for the minimal number of beds, monitoring system, respirators, pacemaker/defibrillators, and necessary additional equipment. The desired function is discussed, namely, the patients to be admitted, the length of stay, and the relocation policy. A uniformed electronic chart for ICCUs is advised, anticipating a common European database. PMID- 15781436 TI - The use of signal-averaged electrocardiogram in risk stratification after acute myocardial infarction in the modern era. PMID- 15781437 TI - Response of respiration of soybean leaves grown at ambient and elevated carbon dioxide concentrations to day-to-day variation in light and temperature under field conditions. AB - BACKGROUND AND AIMS: Respiration is an important component of plant carbon balance, but it remains uncertain how respiration will respond to increases in atmospheric carbon dioxide concentration, and there are few measurements of respiration for crop plants grown at elevated [CO(2)] under field conditions. The hypothesis that respiration of leaves of soybeans grown at elevated [CO(2)] is increased is tested; and the effects of photosynthesis and acclimation to temperature examined. METHODS: Net rates of carbon dioxide exchange were recorded every 10 min, 24 h per day for mature upper canopy leaves of soybeans grown in field plots at the current ambient [CO(2)] and at ambient plus 350 micromol mol( 1) [CO(2)] in open top chambers. Measurements were made on pairs of leaves from both [CO(2)] treatments on a total of 16 d during the middle of the growing seasons of two years. KEY RESULTS: Elevated [CO(2)] increased daytime net carbon dioxide fixation rates per unit of leaf area by an average of 48 %, but had no effect on night-time respiration expressed per unit of area, which averaged 53 mmol m(-2) d(-1) (1.4 micromol m(-2) s(-1)) for both the ambient and elevated [CO(2)] treatments. Leaf dry mass per unit of area was increased on average by 23 % by elevated [CO(2)], and respiration per unit of mass was significantly lower at elevated [CO(2)]. Respiration increased by a factor of 2.5 between 18 and 26 degrees C average night temperature, for both [CO(2)] treatments. CONCLUSIONS: These results do not support predictions that elevated [CO(2)] would increase respiration per unit of area by increasing photosynthesis or by increasing leaf mass per unit of area, nor the idea that acclimation of respiration to temperature would be rapid enough to make dark respiration insensitive to variation in temperature between nights. PMID- 15781438 TI - Optimal hydration status for cryopreservation of intermediate oily seeds: Citrus as a case study. AB - BACKGROUND AND AIMS: The purpose of this study was to investigate the basis of the optimal hydration status for cryopreservation of intermediate oily seeds using Citrus as a model. METHODS: The relationships between equilibrium relative humidity (RH), seed water content, presence of freezable water as determined by DSC analysis, and germination percentage after immersion in liquid nitrogen (LN) were investigated in Citrus aurantifolia, C. grandis, C. madurensis and C. reticulata. The relationship between the lipid content of seeds and their unfrozen water content was also investigated. KEY RESULTS: Independent of their level of seed desiccation tolerance, the optimal desiccation RH for seed tolerance to LN exposure was 75-80 % in the four species studied. This optimal hydration status always coincided with that at which presence of frozen water could not be detected in seed tissues during the cooling/thawing process. The unfrozen water content of seeds was variable between species and negatively correlated to seed lipid content. Using the present data, those obtained previously in seven coffee species and those reported by other authors for five other species, a significant linear relationship was found between the lipid content and the unfrozen water content of seeds. CONCLUSIONS: This study provides additional evidence that intermediate oily seeds do not withstand the presence of freezable water in their tissues during the cooling/warming process. Moreover, it offers two important applied perspectives: (1) independent of their level of desiccation tolerance, testing germination of seeds of a given oily seed species after equilibration in 75-80 % RH at 25 degrees C and LN exposure, gives a rapid and reliable evaluation of the possibility of cryopreserving whole seeds of this given species; (2) it is now possible to calculate the interval of water contents in which non-orthodox oily seeds of a given species are likely to withstand LN exposure as a function of their lipid content. PMID- 15781439 TI - Genetic diversity in fragmented populations of Berchemiella wilsonii var. pubipetiolata (Rhamnaceae). AB - BACKGROUND AND AIMS: [corrected] Berchemiella wilsonii var. pubipetiolata (Rhamnaceae) is distributed in fragmented habitat patches in eastern China. It is highly endangered because of severe disturbance by anthropogenic activities. Information on genetic variation and structure is critical for developing successful conservation strategies for this species. METHODS: Allozyme variation of population genetic diversity and structure was investigated for a total of 98 individuals sampled from four extant populations using isoelectric focusing in thin-layer polyacrylamide slab gels. KEY RESULTS: Based on 20 loci scored from the nine enzymes examined, a high genetic diversity was detected at both the species and population level, while there was a loss of low frequency alleles (<0.1) in all populations. Most loci showed deviation from Hardy-Weinberg equilibrium due to excess of heterozygotes in all populations, suggesting that selection for heterozygotes has occurred in this species. The genetic diversity was mainly found within populations with a moderate genetic differentiation (F(ST) = 0.13), but the two geographically discontinuous population groups showed significant differences, with F-statistic values of 0.078 for the Zhejiang populations and 0.014 for the Anhui populations, respectively. CONCLUSIONS: It appears most likely that this species has experienced a recent decrease in population size, and genetic drift in small populations has resulted in a loss of alleles occurring at low frequency. The differentiation into two population groups reflects a population genetic consequence that has been influenced by the different land-use in the two regions. Some conservation concerns are discussed together with possible strategies for implementing in situ and ex situ conservation. PMID- 15781440 TI - Antioxidant responses of hyper-accumulator and sensitive fern species to arsenic. AB - Plant species capable of hyper-accumulating heavy metals are of considerable interest for phytoremediation, and differ in their ability to accumulate metals from the environment. This work aims to examine (i) arsenic accumulation in three fern species [Chinese brake fern (Pteris vittata L.), slender brake fern (Pteris ensiformis Burm. f.), and Boston fern (Nephrolepis exaltata L.)], which were exposed to 0, 150, or 300 muM of arsenic (Na(2)HAsO(4).7H(2)O), and (ii) the role of anti-oxidative metabolism in arsenic tolerance in these fern species. Arsenic accumulation increased with an increase in arsenic concentration in the growth medium, the most being found in P. vittata fronds showing no toxicity symptoms. In addition, accumulation was highest in the fronds, followed by the rhizome, and finally the roots, in all three fern species. Thiobarbituric acid-reacting substances, indicators of stress in plants, were found to be lowest in P. vittata, which corresponds with its observed tolerance to arsenic. All three ferns responded differentially to arsenic exposure in terms of anti-oxidative defence. Higher levels of superoxide dismutase, catalase, and ascorbate peroxidase were observed in P. vittata than in P. ensiformis and N. exaltata, showing their active involvement in the arsenic detoxification mechanism. However, no significant increase was observed in either guaiacol peroxides or glutathione reductase in arsenic-treated P. vittata. Higher activity of anti oxidative enzymes and lower thiobarbituric acid-reacting substances in arsenic treated P. vittata correspond with its arsenic hyper-accumulation and no symptoms of toxicity. PMID- 15781441 TI - Expression of iron-acquisition-related genes in iron-deficient rice is co ordinately induced by partially conserved iron-deficiency-responsive elements. AB - Rice plants (Oryza sativa L.) utilize the iron chelators known as mugineic acid family phytosiderophores (MAs) to acquire iron from the rhizosphere. Synthesis of MAs and uptake of MA-chelated iron are strongly induced under conditions of iron deficiency. Microarray analysis was used to characterize the expression profile of rice in response to iron deficiency at the genomic level. mRNA extracted from iron-deficient or iron-sufficient rice roots or leaves was hybridized to a rice array containing 8987 cDNA clones. An induction ratio of greater than 2.0 in roots was observed for 57 genes, many of which are involved in iron-uptake mechanisms, including every identified or predicted step in the methionine cycle and the biosynthesis of MAs from methionine. Northern analysis confirmed that the expression of genes encoding every step in the methionine cycle is thoroughly induced by iron deficiency in roots, and almost thoroughly induced in leaves. A promoter search revealed that the iron-deficiency-induced genes related to iron uptake possessed sequences homologous to the iron-deficiency-responsive cis acting elements IDE1 and IDE2 in their promoter regions, at a higher rate than that showing no induction under Fe deficiency. These results suggest that rice genes involved in iron acquisition are co-ordinately regulated by conserved mechanisms in response to iron deficiency, in which IDE-mediated regulation plays a significant role. PMID- 15781442 TI - Up-regulation of the mitochondrial alternative oxidase pathway enhances photosynthetic electron transport under drought conditions. AB - The aim of this study was to explore the role of the mitochondrial alternative oxidase (AOX) in the protection of photosynthesis during drought in wheat leaves. The relative water contents of water-replete and drought-exposed wheat plants were 97.2+/-0.3 and 75+/-2, respectively. Drought increased the amount of leaf AOX protein and also enhanced the rate of AOX-dependent O(2) uptake by the respiratory electron transport chain. The amount of the reduced, active form of the AOX protein was specifically increased by drought. The AOX inhibitor salicylhydroxamic acid (1 mM; SHAM) inhibited 70% of AOX activity in vivo in both water-replete and drought-exposed plants. Plants treated with SHAM were then exposed to low (100), high (350), or excess light (800 mumol photons m(-2) s(-1)) for 90 min. SHAM did not modify chlorophyll a fluorescence quenching parameters in water-replete controls after any of these treatments. However, while the maximal quantum yield of photosystem II (PSII) electron transport (F(v)/F(m)) was not affected by SHAM, the immediate quantum yield of PSII electron transport (Phi(PSII)) and photochemical quenching (qP) were gradually reduced by increasing irradiance in SHAM-treated drought-exposed plants, the decrease being most pronounced at the highest irradiance. Non-photochemical quenching (NPQ) reached near maximum levels in plants subjected to drought at high irradiance. However, a combination of drought and low light caused an intermediate increase in NPQ, which attained higher values when AOX was inhibited. Taken together, these results show that up-regulation of the respiratory AOX pathway protects the photosynthetic electron transport chain from the harmful effects of excess light. PMID- 15781443 TI - Beliefs of Turkish and Moroccan immigrants in The Netherlands about smoking cessation: implications for prevention. AB - Tobacco smoking is a very important preventable cause of mortality and morbidity, and this is also the case in immigrant populations. Therefore, smoking cessation interventions need to take these groups into account. Insight into the applicability of behavioral smoking cessation interventions for non-Western populations is necessary. The objective of our study is to gain insight into the beliefs of smoking cessation in Turkish and Moroccan immigrants in The Netherlands using the I-Change Model. In this model, intention and behavior are supposed to be determined by three types of (psychosocial) factors: attitudes, social influences and self-efficacy expectations. Face-to-face structured interviews among Turkish and Moroccan immigrants were conducted. Results indicate that in contrast to smokers, ex-smokers perceived fewer advantages of smoking and more advantages of smoking cessation. They also perceived less social pressure that encourages smoking (e.g. by being offered cigarettes) and a high self efficacy of being able to quit. The I-Change Model explained 66% of the observed variance. We conclude that the basic factors identified in social cognition theories were replicated in this study. When developing smoking cessation interventions, the results show that it is important to include ethnic-specific salient beliefs, such as the subjective norms of the religious leader. PMID- 15781444 TI - Is personality related to fruit and vegetable intake and physical activity in adolescents? AB - Personality is regarded as a distal determinant of health behavior and has been found to be associated with health behavior in young adults. Research investigating the association between personality and health behaviors in adolescents, however, is lacking. The present study explored the direct associations between Big Five personality dimensions with fruit and vegetable consumption and physical activity (PA) in two Dutch adolescent samples [Sample 1: n = 504; mean age = 14.5 years (SD = 1.7); response rate 20%; Sample 2: n = 476; mean age = 14.9 (SD = 2.0); response rate 17%]. Behavioral outcomes and personality were assessed with validated self-administered questionnaires. Bivariate correlations and multiple regression analyses were conducted, and Cohen's effect sizes were used to interpret the associations found. Agreeableness was positively associated with vegetable consumption, while Openness to Experience was positively associated with fruit consumption and vegetable consumption. Extraversion was positively associated with sports-related PA. Standardized regression coefficients for personality dimensions ranged from 0.09 to 0.13, explaining 3-6% of variance in behavior, indicating small effect sizes. Our results suggest that personality dimensions are associated with fruit and vegetable consumption and sports-related PA in adolescents. Future research should explore possible mediators between personality and health behavior in adolescents. PMID- 15781445 TI - Evaluating healthy schools: perceptions of impact among school-based respondents. AB - Schools are important settings in which to promote children's and young people's physical and emotional health. An evaluation of the National Healthy School Standard in England showed that education and health professionals have implemented a range of projects and activities to improve pupils' health. Although these were generally well received by parents and pupils, they were not uncritical of them. Perceptions of the value of health-related work were influenced by the contextual characteristics of schools--whether primary or secondary, the quality of social relationships, the quality of teaching, and the extent of pupil and parental involvement in the life of the school. With local responsibilities for children's services in England being reorganized in response to the Green Paper, Every Child Matters: Next Steps, there are new opportunities to develop a coherent set of outcome measures that pay due regard to pupils' and parents' views, and which inform collaborative reviews of healthy school programmes, in particular, and local services, more generally. PMID- 15781446 TI - A causal modelling approach to the development of theory-based behaviour change programmes for trial evaluation. AB - Theory-based intervention programmes to support health-related behaviour change aim to increase health impact and improve understanding of mechanisms of behaviour change. However, the science of intervention development remains at an early stage. We present a causal modelling approach to developing complex interventions for evaluation in randomized trials. In this approach a generic model links behavioural determinants, causally through behaviour, to physiological and biochemical variables, and health outcomes. It is tailored to context, target population, behaviours and health outcomes. The development of a specific causal model based on theory and evidence is illustrated by the ProActive programme, supporting increased physical activity among individuals at risk of Type 2 diabetes. The model provides a rational guide to appropriate measures, intervention points and intervention techniques, and can be tested quantitatively. Causal modelling is critically compared to other approaches to intervention development and evaluation, and research directions are indicated. PMID- 15781447 TI - Function of the PEX19-binding site of human adrenoleukodystrophy protein as targeting motif in man and yeast. PMP targeting is evolutionarily conserved. AB - We predicted in human peroxisomal membrane proteins (PMPs) the binding sites for PEX19, a key player in the topogenesis of PMPs, by virtue of an algorithm developed for yeast PMPs. The best scoring PEX19-binding site was found in the adrenoleukodystrophy protein (ALDP). The identified site was indeed bound by human PEX19 and was also recognized by the orthologous yeast PEX19 protein. Likewise, both human and yeast PEX19 bound with comparable affinities to the PEX19-binding site of the yeast PMP Pex13p. Interestingly, the identified PEX19 binding site of ALDP coincided with its previously determined targeting motif. We corroborated the requirement of the ALDP PEX19-binding site for peroxisomal targeting in human fibroblasts and showed that the minimal ALDP fragment targets correctly also in yeast, again in a PEX19-binding site-dependent manner. Furthermore, the human PEX19-binding site of ALDP proved interchangeable with that of yeast Pex13p in an in vivo targeting assay. Finally, we showed in vitro that most of the predicted binding sequences of human PMPs represent true binding sites for human PEX19, indicating that human PMPs harbor common PEX19-binding sites that do resemble those of yeast. Our data clearly revealed a role for PEX19 binding sites as PMP-targeting motifs across species, thereby demonstrating the evolutionary conservation of PMP signal sequences from yeast to man. PMID- 15781448 TI - Effects of protein and gene transfer of the angiopoietin-1 fibrinogen-like receptor-binding domain on endothelial and vessel organization. AB - The vessel-stabilizing effect of angiopoietin-1 (Ang1)/Tie2 receptor signaling is a potential target for pro-angiogenic therapies as well as anti-angiogenic inhibition of tumor growth. We explored the endothelial and vascular specific activities of the Ang1 monomer, i.e. dissociated from its state as an oligomer. A truncated monomeric Ang1 variant (i.e. DeltaAng1) containing the isolated fibrinogen-like receptor-binding domain of Ang1 was created and recombinantly produced in insect cells. DeltaAng1 ligated the Tie2 receptor without triggering its phosphorylation. Moreover, monomeric DeltaAng1 was observed to bind alpha(5)beta(1) integrin with similar affinity compared with Tie2. Unexpectedly, in vitro treatment of endothelial cells with DeltaAng1 showed some of the known effects of full-length Ang1, including inhibition of basal endothelial cell permeability and stimulation of cell adhesion as well as activation of MAPKs. Local treatment of the microvasculature of the developing chicken chorioallantoic membrane with the DeltaAng1 protein led to profound reduction of the mean vascular length density, thinning of vessels, and reduction of the number of vessel branching points. Similar effects were observed in side-by-side experiments with the recombinant full-length Ang1 protein. These effects of simplification of the vessel branching pattern were confirmed through local gene transfer with lentiviral particles encoding DeltaAng1 or full-length Ang1. Together, our findings suggest a potential use for exogenous Ang1 in reducing rather than increasing vascular density. Furthermore, we show that the isolated receptor-binding domain of Ang1 is capable of mediating some effects of full length Ang1 independently of Tie2 phosphorylation, possibly through integrin ligation. PMID- 15781449 TI - Ubiquitination of APOBEC3G by an HIV-1 Vif-Cullin5-Elongin B-Elongin C complex is essential for Vif function. AB - The human immunodeficiency virus type 1 (HIV-1) virion infectivity factor (Vif) overcomes the antiviral activity of APOBEC3G to protect HIV-1 DNA from G-to-A hypermutation. Vif targets APOBEC3G for ubiquitination and proteasomal degradation by forming an SCF-like E3 ubiquitin ligase complex composed of Cullin5, Elongin B, and Elongin C (Vif-BC-Cul5) through a novel SOCS-box motif. In this paper, we have established an in vitro ubiquitin conjugation assay with purified Vif-BC-Cul5 complex and reported that the Vif-BC-Cul5 complex could function as an E3 ligase for APOBEC3G in vitro. A Vif-BC-Cul5 complex promotes the in vitro ubiquitination of the wild type, APOBEC3G but not that of D128K mutant, which does not interact with Vif. We have also investigated several loss of-function Vif mutants. One mutant, SLQ144/146AAA, lost its activity on APOBEC3G because it could not form a complex due to mutations in SOCS-box motif. Other mutants, C114S and C133S, also lost their activity because of loss of the E3 ligase activity of a Vif-BC-Cul5 complex, although these mutants retained the ability to bind to APOBEC3G as well as Cul5 complex. These findings suggest that the E3 ubiquitin ligase activity of the Vif-BC-Cul5 complex is essential for Vif function against APOBEC3G. PMID- 15781451 TI - Nectin-like protein 2 defines a subset of T-cell zone dendritic cells and is a ligand for class-I-restricted T-cell-associated molecule. AB - Dendritic cells (DCs) are a phenotypically and functionally heterogenous population of leukocytes with distinct subsets serving a different set of specialized immune functions. Here we applied an in vitro whole cell panning approach using antibody phage display technology to identify cell-surface epitopes specifically expressed on human blood BDCA3(+) DCs. A single-chain antibody fragment (anti-1F12 scFv) was isolated that recognizes a conserved surface antigen expressed on both human BDCA3(+) DCs and mouse CD8alpha(+) DCs. We demonstrate that anti-1F12 scFv binds Nectin-like protein 2 (Necl2, Tslc1, SynCaM, SgIGSF, or Igsf4), an adhesion molecule involved in tumor suppression, synapse formation, and spermatogenesis. Thus, Necl2 defines a specialized subset of DCs in both mouse and human. We further show that Necl2 binds Class-I restricted T-cell-associated molecule (CRTAM), a receptor primarily expressed on activated cytotoxic lymphocytes. When present on antigen presenting cells, Necl2 regulates IL-22 expression by activated CD8(+) T-cells. We propose that Necl2/CRTAM molecular pair could regulate a large panel of cell/cell interactions both within and outside of the immune system. PMID- 15781450 TI - Bacteriophage T4 helicase loader protein gp59 functions as gatekeeper in origin dependent replication in vivo. AB - Bacteriophage T4 initiates origin-dependent replication via an R-loop mechanism in vivo. During in vitro reactions, the phage-encoded gp59 stimulates loading of the replicative helicase, gp41, onto branched intermediates, including origin R loops. However, although gp59 is essential for recombination-dependent replication from D-loops, it does not appear to be required for origin-dependent replication in vivo. In this study, we have analyzed the origin-replicative intermediates formed during infections that are deficient in gp59 and other phage replication proteins. During infections lacking gp59, the initial replication forks from two different T4 origins actively replicated both leading- and lagging strands. However, the retrograde replication forks from both origins were abnormal in the gp59-deficient infections. The lagging-strand from the initial fork was elongated as a new leading-strand in the retrograde direction without lagging-strand synthesis, whereas in the wild-type, leading- and lagging-strand synthesis appeared to be coupled. These results imply that gp59 inhibits the polymerase holoenzyme in vivo until the helicase-primase (gp41-gp61) complex is loaded, and we thereby refer to gp59 as a gatekeeper. We also found that all origin-replicative intermediates were absent in infections deficient in the helicase gp41 or the single-strand-binding protein gp32, regardless of whether gp59 was present or absent. These results argue that replication from the origin in vivo is dependent on both the helicase and single-strand-binding protein and demonstrate that the strong replication defect of gene 41 and 32 single mutants is not caused by gp59 inhibition of the polymerase. PMID- 15781452 TI - The nature of the stimulus and of the fumarate binding site of the fumarate sensor DcuS of Escherichia coli. AB - DcuS is a membrane-associated sensory histidine kinase of Escherichia coli specific for C(4) -dicarboxylates. The nature of the stimulus and its structural prerequisites were determined by measuring the induction of DcuS-dependent dcuB' 'lacZ gene expression. C(4)-dicarboxylates without or with substitutions at C2/C3 by hydrophilic (hydroxy, amino, or thiolate) groups stimulated gene expression in a similar way. When one carboxylate was replaced by sulfonate, methoxy, or nitro groups, only the latter (3-nitropropionate) was active. Thus, the ligand of DcuS has to carry two carboxylate or carboxylate/nitro groups 3.1-3.8 A apart from each other. The effector concentrations for half-maximal induction of dcuB'-'lacZ expression were 2-3 mm for the C(4)-dicarboxylates and 0.5 mm for 3 nitropropionate or d-tartrate. The periplasmic domain of DcuS contains a conserved cluster of positively charged or polar amino acid residues (Arg(107) X(2)-His(110)-X(9)-Phe(120)-X(26)-Arg(147)-X-Phe(149)) that were essential for fumarate-dependent transcriptional regulation. The presence of fumarate or d tartrate caused sharpening of peaks or chemical shift changes in HSQC NMR spectra of the isolated C(4)-dicarboylate binding domain. The amino acid residues responding to fumarate or d-tartrate were in the region comprising residues 89 150 and including the supposed binding site. DcuS(R147A) mutant with an inactivated binding site was isolated and reconstituted in liposomes. The protein showed the same (activation-independent) kinase activity as DcuS, but autophosphorylation of DcuS was no longer stimulated by C(4)-dicarboxylates. Therefore, the R147A mutation affected signal perception and transfer to the kinase but not the kinase activity per se. PMID- 15781453 TI - Roles of the HIF-1 hypoxia-inducible factor during hypoxia response in Caenorhabditis elegans. AB - The human hypoxia-inducible transcription factor HIF-1 is a critical regulator of cellular and systemic responses to low oxygen levels. When oxygen levels are high, the HIF-1alpha subunit is hydroxylated and is targeted for degradation by the von Hippel-Lindau tumor suppressor protein (VHL). This regulatory pathway is evolutionarily conserved, and the Caenorhabditis elegans hif-1 and vhl-1 genes encode homologs of the HIF-1alpha subunit and VHL. To understand and describe more fully the molecular basis for hypoxia response in this important genetic model system, we compared hypoxia-induced changes in mRNA expression in wild type, hif-1-deficient, and vhl-1-deficient C. elegans using whole genome microarrays. These studies identified 110 hypoxia-regulated gene expression changes, 63 of which require hif-1 function. Mutation of vhl-1 abrogates most hif 1-dependent changes in mRNA expression. Genes regulated by C. elegans hif-1 have predicted functions in signal transduction, metabolism, transport, and extracellular matrix remodeling. We examined the in vivo requirement for 16 HIF-1 target genes and discovered that the phy-2 prolyl 4-hydroxylase alpha subunit is critical for survival in hypoxic conditions. Some HIF-1 target genes negatively regulate formation of stress-resistant dauer larvae. The microarray data presented herein also provide clear evidence for an HIF-1-independent pathway for hypoxia response, and this pathway regulates the expression of multiple heat shock proteins and several transcription factors. PMID- 15781454 TI - Reaction of ferric cytochrome P450cam with peracids: kinetic characterization of intermediates on the reaction pathway. AB - Reactions of substrate-free ferric cytochrome P450cam with peracids to generate Fe=O intermediates have previously been investigated with contradictory results. Using stopped-flow spectrophotometry, the reaction with m-chloroperoxybenzoic acid demonstrated an Fe(IV)=O + porphyrin pi-cation radical (Cpd I) (Egawa, T., Shimada, H., and Ishimura, Y. (1994) Biochem. Biophys. Res. Commun. 201, 1464 1469). By contrast, with peracetic acid, Fe(IV)=O plus a tyrosyl radical were observed by freeze-quench Mossbauer and EPR spectroscopy (Schunemann, V., Jung, C., Trautwein, A. X., Mandon, D., and Weiss, R. (2000) FEBS Lett. 479, 149-154). Our detailed kinetic studies have resolved these contradictory results. At pH >7, a significant fraction of Cpd I is formed transiently, whereas at low pH only a species with a Soret band at 406 nm, presumably Fe(IV)=O + tyrosyl radical, is observed. Evidence for formation of an acylperoxo complex en route to Cpd I was obtained. Because of rapid heme destruction, steps subsequent to formation of the highly oxidized forms could not be fully characterized. Heme destruction was avoided by including peroxidase substrates (e.g. guaiacol), which were oxidized to characteristic peroxidase products as the Fe(III)-P450 was regenerated. Addition of ascorbate to either of the high valent species also reforms the Fe(III) state with only a small loss of heme absorbance. These results indicate that typical peroxidase chemistry occurs with P450cam and offer an explanation for the contrasting results reported earlier. The delineation of improved conditions (pH, temperature, choice of peracid) for generating highly oxidized species with P450cam should be valuable for their further characterization. PMID- 15781455 TI - N-acetylmuramic acid as capping element of alpha-D-fucose-containing S-layer glycoprotein glycans from Geobacillus tepidamans GS5-97T. AB - Geobacillus tepidamans GS5-97(T) is a novel Gram-positive, moderately thermophilic bacterial species that is covered by a glycosylated surface layer (S layer) protein. The isolated and purified S-layer glycoprotein SgtA was ultrastructurally and chemically investigated and showed several novel properties. By SDS-PAGE, SgtA was separated into four distinct bands in an apparent molecular mass range of 106-166 kDa. The three high molecular mass bands gave a positive periodic acid-Schiff staining reaction, whereas the 106-kDa band was nonglycosylated. Glycosylation of SgtA was investigated by means of chemical analyses, 600-MHz nuclear magnetic resonance spectroscopy, and electrospray ionization quadrupole time-of-fight mass spectrometry. Glycopeptides obtained after Pronase digestion revealed the glycan structure [-->2)-alpha-L-Rhap-(1-->3) alpha-D-Fucp-(1-->](n=approximately 20), with D-fucopyranose having never been identified before as a constituent of S-layer glycans. The rhamnose residue at the nonreducing end of the terminal repeating unit of the glycan chain was di substituted. For the first time, (R)-N-acetylmuramic acid, the key component of prokaryotic peptidoglycan, was found in an alpha-linkage to carbon 3 of the terminal rhamnose residue, serving as capping motif of an S-layer glycan. In addition, that rhamnose was substituted at position 2 with a beta-N acetylglucosamine residue. The S-layer glycan chains were bound via the trisaccharide core -->2)-alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1-->3)-alpha-L-Rhap (1--> to carbon 3 of beta-D-galactose, which was attached in O-glycosidic linkage to serine and threonine residues of SgtA of G. tepidamans GS5-97(T). PMID- 15781456 TI - Neuronal expression and neuritogenic action of group X secreted phospholipase A2. AB - Although individual mammalian secreted phospholipase A(2) (sPLA(2)) enzymes exhibit unique tissue and cellular distributions, the cell type-specific functions of each enzyme remain largely unknown. In this study, we found by immunohistochemistry that group X sPLA(2) (sPLA(2)-X) is uniquely located in the peripheral neuronal fibers, an observation that was supported by detection of its transcript and protein in the neuronal cell line PC12 and in primary dorsal root ganglia neurons. Adenoviral expression of sPLA(2)-X in PC12 cells facilitated neurite outgrowth, particularly when combined with a suboptimal concentration of nerve growth factor. In neuronally differentiated PC12 cells, sPLA(2)-X was preferentially localized in the Golgi apparatus and growth cones, and proteolytic conversion of the proenzyme to mature enzyme mainly occurred after the secretion process. The neurite-extending ability of sPLA(2)-X depended on the production of its catalytic product, lysophosphatidylcholine. Moreover, nerve growth factor induced neurite extension of PC12 cells was modestly but significantly attenuated by an anti-sPLA(2)-X antibody or by a small interfering RNA for sPLA(2)-X. These observations suggest the potential contribution of sPLA(2)-X to neuronal differentiation, and possibly repair, under certain conditions. PMID- 15781457 TI - Sp4 is expressed in retinal neurons, activates transcription of photoreceptor specific genes, and synergizes with Crx. AB - To investigate the molecular mechanisms of photoreceptor-specific gene transcription, we examined the role of the neuronal-enriched Sp4 nuclear protein in transcription from the rod-specific beta-PDE and rod opsin gene promoters and compared it to the ubiquitous members of the Sp family, Sp1 and Sp3. Sp4 activates both the rod opsin and beta-PDE promoters, whereas Sp1 activates only the rod opsin promoter and Sp3 activates neither promoter. Interestingly, Sp1 and Sp3 competitively repress Sp4-mediated activation of the beta-PDE promoter. In addition, Sp4, Sp1, and Sp3 each show functional synergy with the photoreceptor enriched Crx transcriptional regulator on the rod opsin promoter but not the beta PDE promoter, although Sp4-mediated activation was the most significant. Sp4, Sp1, and Sp3 bind Crx in co-immunoprecipitation experiments, and their zinc finger domains as well as the Crx homedomain are necessary and sufficient for these interactions. Chromatin immunoprecipitation showed that the rod opsin and beta-PDE promoters are targets of both Sp4 and Crx, which further supports Sp4 Crx interactions in vivo in the context of retinal chromatin environment. In situ hybridization and immunohistochemistry demonstrated that Sp4 is abundantly expressed in various neurons of all retinal layers, and thus co-localizes or overlaps with multiple retina-restricted and -enriched genes, its putative targets. Our results indicate that photoreceptor-specific gene transcription is controlled by the combinatorial action of Sp4 and Crx. The other Sp family members may be involved in photoreceptor-specific transcription directly or through their competition with Sp4. These data suggest the potential importance of Sp4 in retinal neurobiology and pathology. PMID- 15781458 TI - Aspartyl-tRNA synthetase requires a conserved proline in the anticodon-binding loop for tRNA(Asn) recognition in vivo. AB - Most prokaryotes require Asp-tRNA(Asn) for the synthesis of Asn-tRNA(Asn). This misacylated tRNA species is synthesized by a non-discriminating aspartyl-tRNA synthetase (AspRS) that acylates both tRNA(Asp) and tRNA(Asn) with aspartate. In contrast, a discriminating AspRS forms only Asp-tRNA(Asp). Here we show that a conserved proline (position 77) in the L1 loop of the non-discriminating Deinococcus radiodurans AspRS2 is required for tRNA(Asn) recognition in vivo. Escherichia coli trpA34 was transformed with DNA from a library of D. radiodurans aspS2 genes with a randomized codon 77 and then subjected to in vivo selection for Asp-tRNA(Asn) formation by growth in minimal medium. Only proline codons were found at position 77 in the aspS2 genes isolated from 21 of the resulting viable colonies. However, when the aspS temperature-sensitive E. coli strain CS89 was transformed with the same DNA library and then screened for Asp-tRNA(Asp) formation in vivo by growth at the non-permissive temperature, codons for seven other amino acids besides proline were identified at position 77 in the isolates examined. Thus, replacement of proline 77 by cysteine, isoleucine, leucine, lysine, phenylalanine, serine, or valine resulted in mutant D. radiodurans AspRS2 enzymes still capable of forming Asp-tRNA(Asp) but unable to recognize tRNA(Asn). This strongly suggests that proline 77 is responsible for the non-discriminatory tRNA recognition properties of this enzyme. PMID- 15781460 TI - Comprehensive proteomic analysis of Saccharomyces cerevisiae cell walls: identification of proteins covalently attached via glycosylphosphatidylinositol remnants or mild alkali-sensitive linkages. AB - The cell wall of yeast contains proteins that are covalently bound to the glycan network. These cell wall proteins (CWPs) mediate cell-cell interactions and may be involved in cell wall biosynthesis. Using tandem mass spectrometry, we have identified 19 covalently bound CWPs of Saccharomyces cerevisiae. Twelve of them are shown for the first time to be covalently incorporated into the cell wall. The identified proteins include 12 predicted glycosylphosphatidylinositol modified CWPs, all four members of the Pir protein family, and three additional proteins (Scw4p, Scw10p, and Tos1p) that are, like Pir proteins, connected to the cell wall glycan network via an alkali-sensitive linkage. However, Scw4p, Scw10p, and Tos1p do not contain internal repeat sequences shown to be essential for Pir protein incorporation and may represent a separate class of CWPs. Strikingly, seven of the identified proteins (Gas1p, Gas3p, Gas5p, Crh1p, Utr2p, Scw4p, and Scw10p) are classified as glycoside hydrolases. Phenotypic analysis of deletion mutants lacking the corresponding CWP-encoding genes indicated that most of them have altered cell wall properties, which reinforces the importance of the identified proteins for proper cell wall formation. In particular, gas1Delta and ecm33Delta were highly sensitive to Calcofluor White and high temperature, whereas gas1Delta, scw4Delta, and tos1Delta were highly resistant to incubation with beta-1,3-glucanase. The CWP identification method developed here relies on directly generating tryptic peptides from isolated cell walls and is independent of the nature of the covalent linkages between CWPs and cell wall glycans. Therefore, it will probably be equally effective in many other fungi. PMID- 15781461 TI - 2-Haloacrylate reductase, a novel enzyme of the medium chain dehydrogenase/reductase superfamily that catalyzes the reduction of a carbon carbon double bond of unsaturated organohalogen compounds. AB - A soil bacterium, Burkholderia sp. WS, grows on 2-chloroacrylate as the sole carbon source. To identify the enzymes metabolizing 2-chloroacrylate, we carried out comparative two-dimensional gel electrophoresis of the proteins from 2 chloroacrylate- and lactate-grown bacterial cells. As a result, we found that a protein named CAA43 was inducibly synthesized when the cells were grown on 2 chloroacrylate. The CAA43 gene was cloned and shown to encode a protein of 333 amino acid residues (M(r) 35,788) that shared a significant sequence similarity with NADPH-dependent quinone oxidoreductase from Escherichia coli (38.2% identity). CAA43 was overproduced in E. coli and purified to homogeneity. The purified protein catalyzed the NADPH-dependent reduction of the carbon-carbon double bond of 2-chloroacrylate to produce (S)-2-chloropropionate, which is probably further metabolized to (R)-lactate by (S)-2-haloacid dehalogenase in Burkholderia sp. WS. NADH did not serve as a reductant. Despite the sequence similarity to quinone oxidoreductases, CAA43 did not act on 1,4-benzoquinone and 1,4-naphthoquinone. 2-Chloroacrylate analogs, such as acrylate and methacrylate, were also inert as the substrates. In contrast, 2-bromoacrylate served as the substrate. Thus, we named this novel enzyme 2-haloacrylate reductase. This study revealed a new pathway for the degradation of unsaturated organohalogen compounds. It is also notable that the enzyme is useful for the production of (S) 2-chloropropionate, which is used for the industrial production of aryloxyphenoxypropionic acid herbicides. PMID- 15781459 TI - The FOXO3a transcription factor regulates cardiac myocyte size downstream of AKT signaling. AB - Although signaling mechanisms inducing cardiac hypertrophy have been extensively studied, little is known about the mechanisms that reverse cardiac hypertrophy. Here, we describe the existence of a similar Akt/forkhead signaling axis in cardiac myocytes in vitro and in vivo, which is regulated by insulin, insulin like growth factor (IGF), stretch, pressure overload, and angiotensin II stimulation. FOXO3a gene transfer prevented both IGF and stretch-induced hypertrophy in rat neonatal cardiac myocyte cultures in vitro. Transduction with FOXO3a also caused a significant reduction in cardiomyocyte size in mouse hearts in vivo. Akt/FOXO signaling regulated the expression of multiple atrophy-related genes "atrogenes," including the ubiquitin ligase atrogin-1 (MAFbx). In cardiac myocyte cultures, transduction with constitutively active Akt or treatment with IGF suppressed atrogin-1 mRNA expression, whereas transduction with FOXO3a stimulated its expression. FOXO3a transduction activated the atrogin-1 promoter in both cultured myocytes and mouse heart. Thus, in cardiomyocytes, as in skeletal muscle, FOXO3a activates an atrogene transcriptional program, which retards or prevents hypertrophy and is down-regulated by multiple physiological and pathological stimuli of myocyte growth. PMID- 15781462 TI - In vitro synthesized small interfering RNAs elicit RNA interference in african trypanosomes: an in vitro and in vivo analysis. AB - RNA interference (RNAi) describes an epigenetic gene silencing reaction by which gene-specific double-stranded RNA acts as a trigger to induce the ribonucleolytic degradation of homologous transcripts. RNAi in African trypanosomes has been shown to be involved in regulating the transcript abundance of retroposons, and the process currently represents the method of choice in gene function studies of the parasite. However, little is known concerning the mechanistic and structural aspects of the processing reaction. This is in part due to the absence of a trypanosome-specific RNAi in vitro system. Here we demonstrate that both the Dicer and the RNA-induced silencing complex steps of the RNAi reaction pathway can be monitored in vitro using cell-free trypanosome extracts. The two in vitro activities and the generated small interfering RNAs (siRNAs) are characterized by features known from other organisms, and we demonstrate that chemically as well as enzymatically synthesized siRNAs are functional in the parasite. Thus, the transfection of synthetic siRNAs can be used to rapidly monitor gene knockdown phenotypes in Trypanosoma brucei, which should be helpful in genome-wide, RNAi based screening experiments. PMID- 15781464 TI - An immunologically cryptic epitope of Plasmodium falciparum circumsporozoite protein facilitates liver cell recognition and induces protective antibodies that block liver cell invasion. AB - Circumsporozoite, a predominant surface protein, is involved in invasion of liver cells by Plasmodium sporozoites, which leads to malaria. We have previously reported that the amino terminus region (amino acids 27-117) of P. falciparum circumsporozoite protein plays a critical role in the invasion of liver cells by the parasite. Here we show that invasion-blocking antibodies are induced by a polypeptide encoding these 91 amino acids, only when it is presented in the absence of the rest of the protein. This suggests that when present in the whole protein, the amino terminus remains immunologically cryptic. A single reactive epitope was identified and mapped to a stretch of 21 amino acids from position 93 to 113. The epitope is configurational in nature, since its recognition was affected by deleting as little as 3 amino acids from either end of the 21-residue peptide. Lysine 104, the only known polymorphic position in the epitope, affected its recognition by the antibodies, and its conversion to leucine in the protein led to a substantial loss of binding activity of the protein to the hepatocytes. This indicated that in the protein, the epitope serves as a binding ligand and facilitates the interaction between sporozoite and hepatic cells. When considered along with the observation that in its native state this motif is immunologically unresponsive, we suggest that hiding functional moieties of the protein from the immune system is an evasion strategy to preserve liver cell binding function and may be of importance in designing anti-sporozoite vaccines. PMID- 15781463 TI - The Drosophila acetylcholine receptor subunit D alpha5 is part of an alpha bungarotoxin binding acetylcholine receptor. AB - The central nervous system of Drosophila melanogaster contains an alpha bungarotoxin-binding protein with the properties expected of a nicotinic acetylcholine receptor. This protein was purified 5800-fold from membranes prepared from Drosophila heads. The protein was solubilized with 1% Triton X-100 and 0.5 M sodium chloride and then purified using an alpha-cobratoxin column followed by a lentil lectin affinity column. The purified protein had a specific activity of 3.9 micromol of 125I-alpha-bungarotoxin binding sites/g of protein. The subunit composition of the purified receptor was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. This subunit profile was identical with that revealed by in situ labeling of the membrane-bound protein using the photolyzable methyl-4-azidobenzoimidate derivative of 125I-alpha-bungarotoxin. The purified receptor reveals two different protein bands with molecular masses of 42 and 57 kDa. From sedimentation analysis of the purified protein complex in H2O and D2O and gel filtration, a mass of 270 kDa was calculated. The receptor has a s(20,w) of 9.4 and a Stoke's radius of 7.4 nm. The frictional coefficient was calculated to be 1.7 indicating a highly asymmetric protein complex compatible with a transmembrane protein forming an ion channel. The sequence of a peptide obtained after tryptic digestion of the 42-kDa protein allowed the specific identification of the Drosophila D alpha5 subunit by sequence comparison. A peptide-specific antibody raised against the D alpha5 subunit provides further evidence that this subunit is a component of an alpha bungarotoxin binding nicotinic acetylcholine receptor from the central nervous system of Drosophila. PMID- 15781465 TI - Channel function is dissociated from the intrinsic kinase activity and autophosphorylation of TRPM7/ChaK1. AB - TRPM7/ChaK1 is a unique channel/kinase that contains a TRPM channel domain with 6 transmembrane segments fused to a novel serine-threonine kinase domain at its C terminus. The goal of this study was to investigate a possible role of kinase activity and autophosphorylation in regulation of channel activity of TRPM7/ChaK1. Residues essential for kinase activity were identified by site directed mutagenesis. Two major sites of autophosphorylation were identified in vitro by mass spectrometry at Ser(1511) and Ser(1567), and these sites were found to be phosphorylated in intact cells. TRPM7/ChaK1 is a cation-selective channel that exhibits strong outward rectification and inhibition by millimolar levels of internal [Mg(2+)]. Mutation of the two autophosphorylation sites or of a key catalytic site that abolished kinase activity did not alter channel activity measured by whole-cell recording or Ca(2+) influx. Inhibition by internal Mg(2+) was also unaffected in the autophosphorylation site or "kinase-dead" mutants. Moreover, kinase activity was enhanced by Mg(2+), was decreased by Zn(2+), and was unaffected by Ca(2+). In contrast, channel activity was inhibited by all three of these divalent cations. However, deletion of much of C-terminal kinase domain resulted in expression of an apparently inactive channel. We conclude that neither current activity nor regulation by internal Mg(2+) is affected by kinase activity or autophosphorylation but that the kinase domain may play a structural role in channel assembly or subcellular localization. PMID- 15781467 TI - Locating an antagonist in the 5-HT3 receptor binding site using modeling and radioligand binding. AB - We have used a homology model of the extracellular domain of the 5-HT(3) receptor to dock granisetron, a 5-HT(3) receptor antagonist, into the binding site using AUTODOCK. This yielded 13 alternative energetically favorable models. The models fell into 3 groups. In model type A the aromatic rings of granisetron were between Trp-90 and Phe-226 and its azabicyclic ring was between Trp-183 and Tyr 234, in model type B this orientation was reversed, and in model type C the aromatic rings were between Asp-229 and Ser-200 and the azabicyclic ring was between Phe-226 and Asn-128. Residues located no more than 5 A from the docked granisetron were identified for each model; of 26 residues identified, 8 were found to be common to all models, with 18 others being represented in only a subset of the models. To identify which of the docking models best represents the ligand-receptor complex, we substituted each of these 26 residues with alanine and a residue with similar chemical properties. The mutant receptors were expressed in human embryonic kidney (HEK)293 cells and the affinity of granisetron determined using radioligand binding. Mutation of 2 residues (Trp-183 and Glu-129) ablated binding, whereas mutation of 14 other residues caused changes in the [(3)H]granisetron binding affinity in one or both mutant receptors. The data showed that residues both in and close to the binding pocket can affect antagonist binding and overall were found to best support model B. PMID- 15781468 TI - Expression of four glutamine synthetase genes in the early stages of development of rainbow trout (Oncorhynchus mykiss) in relationship to nitrogen excretion. AB - The incorporation of ammonia into glutamine, catalyzed by glutamine synthetase, is thought to be important in the detoxification of ammonia in animals. During early fish development, ammonia is continuously formed as yolk proteins and amino acids are catabolized. We followed the changes in ammonia and urea-nitrogen content, ammonia and urea-nitrogen excretion, glutamine synthetase activity, and mRNA expression of four genes coding for glutamine synthetase (Onmy-GS01-GS04) over 3-80 days post fertilization and in adult liver and skeletal muscle of the rainbow trout (Oncorhynchus mykiss). Both ammonia and urea-nitrogen accumulate before hatching, although the rate of ammonia excretion is considerably higher relative to urea-nitrogen excretion. All four genes were expressed during early development, but only Onmy-GS01 and -GS02 were expressed at appreciable levels in adult liver, and expression was very low in muscle tissue. The high level of expression of Onmy-GS01 and -GS03 prior to hatching corresponded to a linear increase in glutamine synthetase activity. We propose that the induction of glutamine synthetase genes early in development and the subsequent formation of the active protein are preparatory for the increased capacity of the embryo to convert the toxic nitrogen end product, ammonia, into glutamine, which may then be utilized in the ornithine-urea cycle or other pathways. PMID- 15781466 TI - The bone-specific expression of Runx2 oscillates during the cell cycle to support a G1-related antiproliferative function in osteoblasts. AB - The Runx2 (CBFA1/AML3/PEBP2alphaA) transcription factor promotes skeletal cell differentiation, but it also has a novel cell growth regulatory activity in osteoblasts. We addressed here whether Runx2 activity is functionally linked to cell cycle-related mechanisms that control normal osteoblast proliferation and differentiation. We found that the levels of Runx2 gene transcription, mRNA and protein, are each up-regulated with cessation of cell growth (i.e. G(0)/G(1) transition) in preconfluent MC3T3 osteoblastic cells that do not yet express mature bone phenotypic gene expression. Cell growth regulation of Runx2 is also observed in primary calvarial osteoblasts and other osteoblastic cells with relatively normal cell growth characteristics, but not in osteosarcoma cells (e.g. SAOS-2 and ROS17/2.8). Runx2 levels are cell cycle-regulated in MC3T3 cells with respect to the G(1)/S and M/G(1) transitions: oscillates from maximal expression levels during early G(1) to minimal levels during early S phase and mitosis. However, in normal or immortalized (e.g. ATDC5) chondrocytic cells, Runx2 expression is suppressed during quiescence, and Runx2 levels are not regulated during G(1) and S phase in ATDC5 cells. Antisense or small interfering RNA-mediated reduction of the low physiological levels of Runx2 in proliferating MC3T3 cells does not accelerate cell cycle progression. However, forced expression of Runx2 suppresses proliferation of MC3T3 preosteoblasts or C2C12 mesenchymal cells which have osteogenic potential. Forced elevation of Runx2 in synchronized MC3T3 cells causes a delay in G(1). We propose that Runx2 levels and function are biologically linked to a cell growth-related G(1) transition in osteoblastic cells. PMID- 15781469 TI - CHIP controls the sensitivity of transforming growth factor-beta signaling by modulating the basal level of Smad3 through ubiquitin-mediated degradation. AB - Transforming growth factor-beta (TGF-beta) signaling is critical in a variety of biological processes such as cell proliferation, differentiation, and apoptosis. TGF-beta signaling is mediated by a group of proteins including TGF-beta receptors and Smads. It is known that different cells can exhibit different sensitivities to TGF-beta. Several molecular mechanisms, such as the differential expression of the receptor levels, have been suggested as contributing to these differences. Here, we report evidence for a novel mechanism of regulating TGF beta sensitivity that depends on the role of CHIP (carboxyl terminus of Hsc70 interacting protein) in regulating the basal level of Smad3 via the ubiquitin dependent degradation pathway. First, using a luciferase assay we found that overexpression of CHIP inhibited TGF-beta signaling, whereas silencing CHIP expression by small interfering RNAs led to increased TGF-beta signaling sensitivity. Second, based on the results of cell proliferation assays and JunB expression, we found that TGF-beta signaling could be abolished by stably overexpressing CHIP. Third, in those cell lines with stably expressed CHIP, we observed that the Smad3 protein level was dramatically decreased. Finally, we demonstrated that CHIP served as a U-box dependent E3 ligase that can directly mediate ubiquitination and degradation of Smad3 and that this action of CHIP was independent of TGF-beta signaling. Taken together, these findings suggest that CHIP can modulate the sensitivity of the TGF-beta signaling by controlling the basal level of Smad3 through ubiquitin-mediated degradation. PMID- 15781470 TI - Membrane topology of the STT3 subunit of the oligosaccharyl transferase complex. AB - The highly conserved membrane protein STT3 is part of the oligosaccharyl transferase complex in the endoplasmic reticulum of eukaryotic cells. Various experimental observations strongly suggest that STT3 contains the active site of the complex. Here, we report a detailed topology study of STT3 from two different organisms, Saccharomyces cerevisiae and mouse, using in vivo and in vitro topology mapping assays. Our results suggest that STT3 has 11 transmembrane helices and an overall N(cyt)-C(lum) orientation. PMID- 15781471 TI - Thiazide-sensitive NaCl-cotransporter in the intestine: possible role of hydrochlorothiazide in the intestinal Ca2+ uptake. AB - Thiazides, such as hydrochlorothiazide (HCTZ), are used to control blood pressure and to reduce renal calcium excretion. These effects are a result of interactions with the NaCl-cotransporter (NCC). This is demonstrated by the fact that mutations within the NCC protein lead to salt-resistant hypotension and hypocalciuria, paralleled by an increase in bone mineral density. These symptoms are also known as Gitelman syndrome. It has become increasingly evident that the effect of HCTZ on blood pressure and calcium homeostasis cannot be attributed exclusively to kidney functions, where the primary action of HCTZ on NCC is postulated to occur. We demonstrated the presence of the NCC transporter in the rat small intestine (ileum and jejunum) and human HT-29 cells, by using reverse transcription-PCR, Northern blot, Western blot, and immunofluorescence. Furthermore, we show that HCTZ modulates Ca(2+) uptake by intestinal cells, while affecting the electrical parameters of the cellular membrane, thus suggesting a functional interaction between NCC and the epithelial voltage-dependent calcium channel. The experiments presented here support the hypothesis of a direct involvement of the intestinal cells in the interaction between HCTZ and NaCl, as well as calcium homeostasis. PMID- 15781472 TI - Roles of the N-terminal domain on the function and quaternary structure of the ionotropic glutamate receptor. AB - The alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid (AMPA) subtype of ionotropic glutamate receptors (iGluRs) mediates fast excitatory neurotransmission in the mammalian brain. Although the most N-terminal leucine/isoleucine/valine-binding protein (LIVBP) domain is suggested to play a role in the initial assembly of iGluR subunits, it is unclear how this domain is arranged and functions in intact iGluRs. Similarly, although recent crystallographic analyses indicate that the isolated ligand-binding lysine/arginine/ornithine-binding protein domain forms a 2-fold symmetric dimer, the subunit stoichiometry of intact iGluRs remains elusive. Here, we developed a new approach to address these issues. The LIVBP domain of the GluR1 subunit of AMPA receptors was replaced by leucine-zipper peptides designed to form stable symmetric dimers, trimers, tetramers, or pentamers. All these mutant GluR1s were expressed in human embryonic kidney 293 cells and were transported to the cell surface as well as wild type GluR1. Functional and biochemical analyses indicated that these oligomerizing peptides specifically controlled the formation of the expected number of subunits in a channel complex. However, the channel function was only restored by the tetramer-forming peptide. Although the purified LIVBP domain of GluR1 formed a dimmer in solution, a dimer-forming peptide could not restore the function of GluR1. Moreover, a cross-linking assay indicated that four LIVBP domains are located in proximity to each other. These results suggest that the function of the LIVBP domain is not simply to form initial dimers but to adopt a conformation compatible with the overall tetrameric arrangement of subunits in intact AMPA receptors. PMID- 15781473 TI - Fibroblast growth factor (FGF) 18 signals through FGF receptor 3 to promote chondrogenesis. AB - Signaling by fibroblast growth factor (FGF) 18 and FGF receptor 3 (FGFR3) have been shown to regulate proliferation, differentiation, and matrix production of articular and growth plate chondrocytes in vivo and in vitro. Notably, the congenital absence of either FGF18 or FGFR3 resulted in similar expansion of the growth plates of fetal mice and the addition of FGF18 to human articular chondrocytes in culture enhanced proliferation and matrix production. Based on these and other experiments it has been proposed that FGF18 signals through FGFR3 to promote cartilage production by chondrocytes. Its role in chondrogenesis remains to be defined. In the current work we used the limb buds of FGFR3(+/+) and FGFR3(-/-) embryonic mice as a source of mesenchymal cells to determine how FGF18 signaling affects chondrogenesis. Confocal laser-scanning microscopy demonstrated impaired cartilage nodule formation in the FGFR3(-/-) cultures. Potential contributing factors to the phenotype were identified as impaired mitogenic response to FGF18, decreased production of type II collagen and proteoglycan in response to FGF18 stimulation, impaired interactions with the extracellular matrix resulting from altered integrin receptor expression, and altered expression of FGFR1 and FGFR2. The data identified FGF18 as a selective ligand for FGFR3 in limb bud mesenchymal cells, which suppressed proliferation and promoted their differentiation and production of cartilage matrix. This work, thus, identifies FGF18 and FGFR3 as potential molecular targets for intervention in tissue engineering aimed at cartilage repair and regeneration of damaged cartilage. PMID- 15781474 TI - TGF-{beta}1 activates two distinct type I receptors in neurons: implications for neuronal NF-{kappa}B signaling. AB - Transforming growth factor-betas (TGF-betas) are pleiotropic cytokines involved in development and maintenance of the nervous system. In several neural lesion paradigms, TGF-beta1 exerts potent neuroprotective effects. Neurons treated with TGF-beta1 activated the canonical TGF-beta receptor I/activin-like kinase receptor 5 (ALK5) pathway. The transcription factor nuclear factor-kappaB (NF kappaB) plays a fundamental role in neuroprotection. Treatment with TGF-beta1 enhanced NF-kappaB activity in gelshift and reporter gene analyses. However, ectopic expression of a constitutively active ALK5 failed to mimic these effects. ALK1 has been described as an alternative TGF-beta receptor in endothelial cells. Interestingly, we detected significant basal expression of ALK1 and its injury induced up-regulation in neurons. Treatment with TGF-beta1 also induced a pronounced increase in downstream Smad1 phosphorylation. Overexpression of a constitutively active ALK1 mimicked the effect of TGF-beta1 on NF-kappaB activation and neuroprotection. Our data suggest that TGF-beta1 simultaneously activates two distinct receptor pathways in neurons and that the ALK1 pathway mediates TGF-beta1-induced NF-kappaB survival signaling. PMID- 15781475 TI - Hrp59, an hnRNP M protein in Chironomus and Drosophila, binds to exonic splicing enhancers and is required for expression of a subset of mRNAs. AB - Here, we study an insect hnRNP M protein, referred to as Hrp59. Hrp59 is relatively abundant, has a modular domain organization containing three RNA binding domains, is dynamically recruited to transcribed genes, and binds to premRNA cotranscriptionally. Using the Balbiani ring system of Chironomus, we show that Hrp59 accompanies the mRNA from the gene to the nuclear envelope, and is released from the mRNA at the nuclear pore. The association of Hrp59 with transcribed genes is not proportional to the amount of synthesized RNA, and in vivo Hrp59 binds preferentially to a subset of mRNAs, including its own mRNA. By coimmunoprecipitation of Hrp59-RNA complexes and microarray hybridization against Drosophila whole-genome arrays, we identify the preferred mRNA targets of Hrp59 in vivo and show that Hrp59 is required for the expression of these target mRNAs. We also show that Hrp59 binds preferentially to exonic splicing enhancers and our results provide new insights into the role of hnRNP M in splicing regulation. PMID- 15781476 TI - Targeting of Arf-1 to the early Golgi by membrin, an ER-Golgi SNARE. AB - Arf and Rab family GTPases regulate membrane traffic in cells, yet little is known about how they are targeted to distinct organelles. To identify sequences in Arf-1 necessary for Golgi targeting, we examined the localization of chimeras between Arf-1 and Arf-6. Here, we identify a 16-amino acid sequence in Arf-1 that specifies Golgi targeting and contains a motif (MXXE) that is important for Arf-1 binding to membrin, an ER-Golgi SNARE protein. The MXXE motif is conserved in all Arfs known to localize to the Golgi and enables Arf-1 to localize to the early Golgi. Arf-1 lacking these 16 aa can still localize to the late Golgi where it displays a more rapid Golgi-cytosol cycle than wild-type Arf-1. These studies suggest that membrin recruits Arf-1 to the early Golgi and reveal distinct kinetic cycles for Arf-1 at early and late Golgi determined by different sets of Arf regulators and effectors. PMID- 15781478 TI - Golgi positioning: are we looking at the right MAP? AB - One of the characteristics of the mammalian Golgi is its position adjacent to the nucleus. This characteristic is maintained through the action of the microtubule (MT) minus end-directed motor dynein and MT-associated proteins (MAPs). Recent findings suggest that GMAP-210, a member of the golgin family of proteins, may help to link Golgi membranes and vesicles with the MT cytoskeleton. However, there are good grounds to doubt that either GMAP-210 or its yeast homologue Rud3p is a MAP. Instead, they appear to function in vesicle trafficking events at the Golgi together with the GTPase ARF1 and a small membrane protein, Erv14. As such, the interesting question of how the Golgi interacts with MTs may well remain open to further investigation. PMID- 15781479 TI - Idiopathic urinary stone disease: possible polygenic aetiological factors. PMID- 15781477 TI - Sox2 induction by FGF and FGFR2 activating mutations inhibits Wnt signaling and osteoblast differentiation. AB - Activating mutations in fibroblast growth factor receptor 2 (FGFR2) cause several craniosynostosis syndromes by affecting the proliferation and differentiation of osteoblasts, which form the calvarial bones. Osteoblasts respond to FGF with increased proliferation and inhibition of differentiation. We analyzed the gene expression profiles of osteoblasts expressing FGFR2 activating mutations (C342Y or S252W) and found a striking down-regulation of the expression of many Wnt target genes and a concomitant induction of the transcription factor Sox2. Most of these changes could be reproduced by treatment of osteoblasts with exogenous FGF. Wnt signals promote osteoblast function and regulate bone mass. Sox2 is expressed in calvarial osteoblasts in vivo and we show that constitutive expression of Sox2 inhibits osteoblast differentiation and causes down-regulation of the expression of numerous Wnt target genes. Sox2 associates with beta-catenin in osteoblasts and can inhibit the activity of a Wnt responsive reporter plasmid through its COOH-terminal domain. Our results indicate that FGF signaling could control many aspects of osteoblast differentiation through induction of Sox2 and regulation of the Wnt-beta-catenin pathway. PMID- 15781480 TI - Democracy and health. PMID- 15781481 TI - Recent advances in the monitoring and management of diabetic ketoacidosis. PMID- 15781482 TI - Refeeding syndrome: life-threatening, underdiagnosed, but treatable. PMID- 15781483 TI - Recent advances in the monitoring and management of diabetic ketoacidosis. PMID- 15781484 TI - The obesity epidemic. PMID- 15781485 TI - Dialogue and diagnosis. PMID- 15781486 TI - Adjuvant therapy for colon cancer: 12 months, 9 months, 6 months ... why not 3 months? PMID- 15781487 TI - Timing of quality of life assessment in cancer clinical trials: fine tuning remains a challenge. PMID- 15781489 TI - Hematopoietic stem cell transplantation in mantle cell lymphoma. AB - BACKGROUND: Patients with mantle cell lymphoma (MCL) have in general, lower response rates and overall survival (OS) than those with other B-cell non Hodgkin's lymphomas. The role of hematopoietic stem cell transplantation (HSCT) in MCL is unclear. Hence we decided to study the clinical course of patients who received autologous and allogeneic HSCT for MCL. METHODS: Ninety-seven patients, (80 patients-autologous; 17 patients-allogeneic) who received a HSCT for mantle cell lymphoma were included in the study. RESULTS: The complete response rates at day 100 between the two groups were similar (73% vs. 62%). Day-100 mortality was higher in the allogeneic HSCT group (19% vs. 0%) (P < 0.01). The estimated 5-year relapse rates, 5-year event-free survival (EFS) and 5-year OS among the allogeneic HSCT patients were 21%, 44% and 49%, respectively, similar to 56%, 39% and 47% in the autologous group. Ten patients received HyperCVAD (hyperfractionated cyclophosphamide, vincristine, doxorubicin and dexamethasone + high-dose methotrexate and cytarabine) +/- rituximab prior to transplant. There have been no relapses or deaths amongst these patients at a median follow-up of 16 months. CONCLUSIONS: Patients treated with allogeneic HSCT had a lower relapse rate, but similar EFS and OS to autologous HSCT. Treatment of MCL with HyperCVAD +/- rituximab followed by HSCT seems promising. PMID- 15781488 TI - Consensus meeting for the management of gastrointestinal stromal tumors. Report of the GIST Consensus Conference of 20-21 March 2004, under the auspices of ESMO. AB - BACKGROUND: The management of gastrointestinal stromal tumors (GIST) has evolved very rapidly in the last 4 years. The objectives of this international consensus meeting were to describe the optimal management procedures for patients with GIST in localized and advanced stages, as well as research issues for the future. MATERIALS AND METHODS: A panel of experts from six specialties, including pathology, molecular biology, imaging, surgery, medical oncology and methodologists for clinical practice guidelines from different European and extra European sarcoma societies were invited to a 2-day workshop. Several questions were selected by the organizing committee prior to the conference. Selected panelists reviewed the current levels of evidence for each point, and presented their conclusions during the meeting. These proposals were discussed, and consensus points were identified and categorized according to the Standard Options Recommandations (SOR) of the French Federation of Cancer Centers and National Comprehensive Cancer Network (NCCN). RESULTS: Thirty-two consensus points were identified, most from categories 2A of the NCCN and B2 of the SOR. Among these, the standard histological examination with immunohistochemical analysis using CD117, CD34, PS100, desmin and smooth muscle actin is considered standard. Molecular biology for the identification of KIT and PDGFRA mutation is an optional diagnostic procedure for GIST with negative CD117 staining, and otherwise is considered a research procedure. Complete tumor resection with negative tumor margins is the standard surgical treatment. Adjuvant imatinib after optimal tumor resection as well as neo-adjuvant imatinib remain experimental approaches to be performed within prospective clinical studies. Imatinib should be started at the date of diagnosis of metastatic relapse and given until development of intolerance or progressive disease. The optimal criteria for tumor response to imatinib remain to be delineated, and should include not only tumor size reduction or disease stabilization, but also reduction of tumor density (Hounsfield Units) on computed tomography and metabolic activity (i.e. reduction of FDG uptake on positron emission tomography). In a substantial proportion of patients, stable disease and even increase in tumor size may be associated with pathologic response to imatinib therapy, and available survival data indicate that the survival of these patients is similar to that of patients with conventional tumor response. Metastasis resection is an experimental procedure. CONCLUSIONS: Consensus points in clinical management of GIST as well as questions for future clinical trials were identified during this consensus conference on GIST management. PMID- 15781491 TI - Modification at position 9 with 1-methyladenosine is crucial for structure and function of nematode mitochondrial tRNAs lacking the entire T-arm. AB - The mitochondria of the nematode Ascaris suum have tRNAs with unusual secondary structures that lack either the T-arm or D-arm found in most other organisms. Of the twenty-two tRNA species present in the mitochondria of A.suum, twenty lack the entire T-arm and two serine tRNAs lack the D-arm. To understand how such unusual tRNAs work in the nematode mitochondrial translation system, we analyzed post-transcriptional modifications of 11 mitochondrial tRNA species purified from A.suum, 10 of which lacked a T-arm and one of which lacked a D-arm. The most characteristic feature of nematode mitochondrial tRNAs lacking a T-arm was the presence of 1-methyladenosine at position 9 (m1A9). Synthesis of T-armless tRNAs with or without the modified nucleoside showed that T-armless tRNAs without the modification had much lower aminoacylation and EF-Tu-binding activities than native tRNAs. The addition of a single methyl group to A9 of these tRNAs was sufficient to restore nearly native levels of aminoacylation and EF-Tu-binding activity as well as tertiary structure, suggesting that m1A9 is a key residue for the activity of T-armless tRNAs. Thus, m1A9 is indispensable for the structure and function of T-armless tRNAs of nematode mitochondrial origin. PMID- 15781492 TI - Single-stranded DNA-binding protein of Deinococcus radiodurans: a biophysical characterization. AB - The highly conserved bacterial single-stranded DNA-binding (SSB) proteins play an important role in DNA replication, repair and recombination and are essential for the survival of the cell. They are functional as tetramers, in which four OB(oligonucleotide/oligosaccharide binding)-folds act as DNA-binding domains. The protomer of the SSB protein from the extremely radiation-resistant organism Deinococcus radiodurans (DraSSB) has twice the size of the other bacterial SSB proteins and contains two OB-folds. Using analytical ultracentrifugation, we could show that DraSSB forms globular dimers with some protrusions. These DraSSB dimers can interact with two molecules of E.coli DNA polymerase III chi subunit. In fluorescence titrations with poly(dT) DraSSB bound 47-54 nt depending on the salt concentration, and fluorescence was quenched by more than 75%. A distinct low salt binding mode as for EcoSSB was not observed for DraSSB. Nucleic acid binding affinity, rate constant and association mechanism are quite similar for EcoSSB and DraSSB. In a complementation assay in E.coli, DraSSB took over the in vivo function of EcoSSB. With DraSSB behaving almost identical to EcoSSB the question remains open as to why dimeric SSB proteins have evolved in the Thermus group of bacteria. PMID- 15781493 TI - A systematic analysis of the silencing effects of an active siRNA at all single nucleotide mismatched target sites. AB - The specificity of small interfering RNA (siRNA)-mediated gene silencing is a critical consideration for the application of RNA interference (RNAi). While the discovery of potential off-target effects by siRNAs is of concern, no systematic analysis has been conducted to explore the specificity of RNAi. Here, we present a study where a functionally validated siRNA (siCD46) was examined for silencing specificity on all possible 57 permutated target sites, each carrying a single nucleotide mutation that would generate a mismatch when paired with siRNA antisense strand. We found that it was not only the position of the mismatched base pair, but also the identity of the nucleotides forming the mismatch that influenced silencing. Surprisingly, mismatches formed between adenine (A) and cytosine (C), in addition to the G:U wobble base pair, were well tolerated and target sites containing such mismatches were silenced almost as efficiently as its fully matched counterpart by siCD46. Northern blots showed that the silencing of fusion genes harboring the mutated target sites involved target mRNA degradation. This study provides direct evidence that the target recognition of siRNA is far more degenerative than previously considered. This finding is instrumental in the understanding of RNAi specificity and may aid the computational prediction of RNA secondary structure. PMID- 15781494 TI - Both RNase E and RNase III control the stability of sodB mRNA upon translational inhibition by the small regulatory RNA RyhB. AB - Previous work has demonstrated that iron-dependent variations in the steady-state concentration and translatability of sodB mRNA are modulated by the small regulatory RNA RyhB, the RNA chaperone Hfq and RNase E. In agreement with the proposed role of RNase E, we found that the decay of sodB mRNA is retarded upon inactivation of RNase E in vivo, and that the enzyme cleaves within the sodB 5' untranslated region (5'-UTR) in vitro, thereby removing the 5' stem-loop structure that facilitates Hfq and ribosome binding. Moreover, RNase E cleavage can also occur at a cryptic site that becomes available upon sodB 5'-UTR/RyhB base pairing. We show that while playing an important role in facilitating the interaction of RyhB with sodB mRNA, Hfq is not tightly retained by the RyhB-sodB mRNA complex and can be released from it through interaction with other RNAs added in trans. Unlike turnover of sodB mRNA, RyhB decay in vivo is mainly dependent on RNase III, and its cleavage by RNase III in vitro is facilitated upon base pairing with the sodB 5'-UTR. These data are discussed in terms of a model, which accounts for the observed roles of RNase E and RNase III in sodB mRNA turnover. PMID- 15781495 TI - The genome sequence of Salmonella enterica serovar Choleraesuis, a highly invasive and resistant zoonotic pathogen. AB - Salmonella enterica serovar Choleraesuis (S. Choleraesuis), a highly invasive serovar among non-typhoidal Salmonella, usually causes sepsis or extra-intestinal focal infections in humans. S. Choleraesuis infections have now become particularly difficult to treat because of the emergence of resistance to multiple antimicrobial agents. The 4.7 Mb genome sequence of a multidrug resistant S. Choleraesuis strain SC-B67 was determined. Genome wide comparison of three sequenced Salmonella genomes revealed that more deletion events occurred in S. Choleraesuis SC-B67 and S.Typhi CT18 relative to S. Typhimurium LT2. S. Choleraesuis has 151 pseudogenes, which, among the three Salmonella genomes, include the highest percentage of pseudogenes arising from the genes involved in bacterial chemotaxis signal-transduction pathways. Mutations in these genes may increase smooth swimming of the bacteria, potentially allowing more effective interactions with and invasion of host cells to occur. A key regulatory gene of TetR/AcrR family, acrR, was inactivated through the introduction of an internal stop codon resulting in overexpression of AcrAB that appears to be associated with ciprofloxacin resistance. While lateral gene transfer providing basic functions to allow niche expansion in the host and environment is maintained during the evolution of different serovars of Salmonella, genes providing little overall selective benefit may be lost rapidly. Our findings suggest that the formation of pseudogenes may provide a simple evolutionary pathway that complements gene acquisition to enhance virulence and antimicrobial resistance in S. Choleraesuis. PMID- 15781497 TI - Hemoglobin-based oxygen carrying solutions: will they replace red blood cells? PMID- 15781498 TI - Systemic responses to hemodilution after transfusion with stored blood and with a hemoglobin-based oxygen carrier. AB - We assessed the systemic effects of exchanges with blood or hemoglobin (Hb) raffimer under conditions of critical oxygen delivery (Do(2)crit). We compared Do(2)crit in animals receiving Hb-based oxygen carrier (HBOC; Hemolink), fresh blood (collected <24 h), or stored blood (10 days) before hemodilution. Rats were randomized to control, blood, or HBOC isovolemic exchange. Oxygen consumption was measured by using expired gas (o(2)a) and blood (o(2)b) samples, whereas whole body oxygen delivery (Do(2)) was calculated from cardiac output and arterial oxygen content. After exchange, rats were subjected to stepwise isovolemic hemodilution. Blood pressure, gases, acid-base status, glucose, Hb oxygen saturation, heart rate, and total peripheral resistance were also measured. We found that 1) HBOC-treated rats showed an increased mean arterial blood pressure and total peripheral resistance throughout the hemodilution, 2) Do(2)crit calculated with o(2)a or o(2)b gave identical results, 3) Do(2)crit was not different between animals receiving blood and those receiving HBOC, 4) the terminal Hb concentration (1.8 +/- 0.1 g/dL) and Do(2) (5 +/- 1 mL . min(-1) . kg(-1)) were similar for all animals, and 5) most oxygen transport and biochemical variables changed similarly during hemodilution. The data suggest that tolerance to Do(2)crit is not altered by 50% replacement of native Hb by stored blood or Hb raffimer. PMID- 15781499 TI - Epidural anesthesia for coronary artery bypass surgery compared with general anesthesia alone does not reduce biochemical markers of myocardial damage. AB - High thoracic epidural anesthesia/analgesia (HTEA) for coronary artery bypass grafting (CABG) surgery may have myocardial protective effects. In this prospective randomized controlled study, we investigated the effect of HTEA for elective CABG surgery on the release of troponin I, time to tracheal extubation, and analgesia. One-hundred-twenty patients were randomized to a general anesthesia (GA) group or a GA plus HTEA group. The GA group received fentanyl (7 15 microg/kg) and a morphine infusion. The HTEA group received fentanyl (5-7 microg/kg) and an epidural infusion of ropivacaine 0.2% and fentanyl 2 microg/mL until postoperative Day 3. There were no differences in troponin I levels between study groups. The time to tracheal extubation [median (interquartile range)] in the HTEA group was 15 min (10-320 min), compared with 430 min (284-590 min) in the GA group (P < 0.0001). Analgesia was improved in the HTEA group compared with the GA group. Mean arterial blood pressure poststernotomy and systemic vascular resistance in the intensive care unit were lower in the HTEA group. We conclude that HTEA for CABG surgery had no effect on troponin release but improved postoperative analgesia and was associated with a reduced time to extubation. PMID- 15781500 TI - Propofol attenuates lung endothelial injury induced by ischemia-reperfusion and oxidative stress. AB - Lung dysfunction after cardiopulmonary bypass and lung transplantation results from oxidant-mediated cellular damage. Previously, we observed the shedding of angiotensin-converting enzyme (ACE) from the endothelial cell surface to be a more sensitive and earlier marker of oxidative lung endothelial injury than lung wet-to-dry weight ratio. The aim of this study was to evaluate the potential of the anesthetic propofol, which has antioxidant properties, to prevent oxidative lung injury by measuring ACE shedding. ACE release from isolated perfused rat lungs increased significantly after ischemia-reperfusion (I/R). Propofol significantly decreased I/R-induced ACE release by 23.4% (P < 0.05). Perfusion with 0.75 mM H(2)O(2) also caused ACE release from the lung microvasculature, which was similarly attenuated by propofol. The protective effect of propofol on H(2)O(2)-induced ACE shedding was confirmed in vitro using Chinese Hamster Ovary cells overexpressing human ACE. Thus, propofol can attenuate oxidative injury of the pulmonary endothelium as detected by ACE shedding in I/R and H(2)O(2) models of acute lung injury. PMID- 15781496 TI - Formation of linear inverted repeat amplicons following targeting of an essential gene in Leishmania. AB - Attempts to inactivate an essential gene in the protozoan parasite Leishmania have often led to the generation of extra copies of the wild-type alleles of the gene. In experiments with Leishmania tarentolae set up to disrupt the gene encoding the J-binding protein 1 (JBP1), a protein binding to the unusual base beta-D-glucosyl-hydroxymethyluracil (J) of Leishmania, we obtained JBP1 mutants containing linear DNA elements (amplicons) of approximately 100 kb. These amplicons consist of a long inverted repeat with telomeric repeats at both ends and contain either the two different targeting cassettes used to inactivate JBP1, or one cassette and one JBP1 gene. Each long repeat within the linear amplicons corresponds to sequences covering the JBP1 locus, starting at the telomeres upstream of JBP1 and ending in a approximately 220 bp sequence repeated in an inverted (palindromic) orientation downstream of the JBP1 locus. We propose that these amplicons have arisen by a template switch inside a DNA replication fork involving the inverted DNA repeats and helped by the gene targeting. PMID- 15781501 TI - Usefulness of unipolar epicardial ventricular electrogram for triggering of intraaortic balloon counterpulsation during off-pump coronary artery bypass surgery in patients with hemodynamic instability complicating acute coronary syndromes. AB - During off-pump coronary artery bypass graft (CABG) surgery for patients requiring preoperative intraaortic balloon counterpulsation (IABP) support, errors in timing of IABP during mobilization of the heart can induce unstable hemodynamic conditions. We applied a simple technique for triggering IABP accurately during off-pump CABG: one end of an epicardial pacemaker wire is placed on the surface of the left ventricle, and the other end is linked to one of the precordial V leads. This technique provided an exact detection of R-wave potentials in any position of the heart during the off-pump CABG procedure. Ten patients with hemodynamic instability complicating acute coronary syndromes underwent off-pump CABG with this technique, including grafting to the circumflex system, safely and successfully. PMID- 15781502 TI - Assessing fluid-responsiveness by a standardized ventilatory maneuver: the respiratory systolic variation test. AB - Respiratory-induced changes in arterial blood pressure predict fluid responsiveness. However, the accuracy of these variables is affected by the preset tidal volume and by the early inspiratory increase in arterial blood pressure. We have therefore calculated the slope produced by the minimal systolic blood pressures (plotted against the respective airway pressures) during a ventilatory maneuver consisting of four incremental, successive, pressure controlled breaths, termed the Respiratory Systolic Variation Test (RSVT). In 14 ventilated patients, after major vascular surgery, the slope of the RSVT decreased significantly after intravascular fluid administration and correlated with the end-diastolic area and with changes in cardiac output better than filling pressures. This preliminary study suggests that a standardized ventilatory maneuver may be useful in guiding fluid therapy in ventilated patients. PMID- 15781503 TI - The use of argatroban for carotid endarterectomy in heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a major obstacle in cardiovascular surgeries. In this case report, we used argatroban, a direct thrombin inhibitor, to achieve and maintain anticoagulation for carotid endarterectomy. Unlike heparin, the direct thrombin inhibitors bind directly to thrombin, bypassing antithrombin III and the potential to precipitate HIT. A bolus of argatroban 150 microg/kg followed by an infusion of 5 microg . kg(-1) . min(-1) was used, and adequate anticoagulation was demonstrated with multiple laboratory tests (at 28 min, prothrombin time = 29.8 s, partial thromboplastin time = 69.1 s, international normalized ratio = 3.52 s, and activated clotting time = 220 s). The surgery was successful, and the patient was discharged the next day with no postoperative neurologic sequelae or other complications. We conclude that argatroban can be used safely and successfully for carotid endarterectomy in a patient with a history of HIT. PMID- 15781504 TI - The endoscopically measured effects of airway maneuvers and the lateral position on airway patency in anesthetized children with adenotonsillar hypertrophy. AB - Obstruction of the upper airway is a major challenge for anesthesiologists administering general anesthesia in spontaneously breathing children with adenotonsillar hypertrophy. Lateral positioning is a simple treatment for obstructive sleep apnea. In this study, we examined the effects of body position shifting and common airway maneuvers such as chin lift and jaw thrust on airway patency (stridor score and upper airway dimensions by endoscopy) in anesthetized children scheduled for adenotonsillectomy. Eighteen children aged 1-11 yr were anesthetized with sevoflurane. During spontaneous breathing with 5% sevoflurane and 100% oxygen, upper airway dimensions and stridor score were recorded. After baseline recording, chin lift and jaw thrust were performed in both the supine and the lateral decubitus position. Chin lift, jaw thrust, and lateral position increased the airway dimensions and improved the stridor score. Moreover, lateral positioning enhanced the effects of these airway maneuvers on airway patency. We concluded that lateral positioning combined with airway maneuvers provided better airway patency for anesthetized children with adenotonsillar hypertrophy. PMID- 15781505 TI - The effects of a small-dose naloxone infusion on opioid-induced side effects and analgesia in children and adolescents treated with intravenous patient-controlled analgesia: a double-blind, prospective, randomized, controlled study. AB - Opioids are frequently associated with side effects such as nausea, vomiting, and pruritus. We hypothesized that a prophylactic, continuous small-dose naloxone infusion would reduce the incidence of opioid-induced side effects without affecting analgesia or opioid consumption. In this prospective, double-blind, randomized, controlled clinical trial, we studied 46 postoperative patients (M:F, 21:25), averaging 14 +/- 2.5 yr and 53 +/- 17 kg, at the start of morphine IV patient-controlled analgesia. Patients were randomized to either saline (control, n = 26) or naloxone 0.25 microg . kg(-1) . h(-1) (n = 20). We found that the incidence and severity of pruritus (77% versus 20%; P < 0.05) and nausea (70% versus 35%; P < 0.05) was significantly more frequent in the placebo group compared with the naloxone group. Morphine consumption (1.02 +/- 0.41 mg . kg(-1) . d(-1) versus 1.28 +/- 0.61 mg . kg(-1) . d(-1)), pain scores at rest (4 +/- 2 versus 3 +/- 2), and pain scores with coughing (6 +/- 2 versus 6 +/- 2) were not different. We conclude that, in children and adolescents, a small-dose naloxone infusion (0.25 microg . kg(-1) . h(-1)) can significantly reduce the incidence and severity of opioid-induced side effects without affecting opioid-induced analgesia. When initiating morphine IV patient-controlled analgesia for the treatment of moderate to severe pain, clinicians should strongly consider starting a concomitant small-dose naloxone infusion. PMID- 15781506 TI - Recovery after anesthesia for short pediatric oncology procedures: propofol and remifentanil compared with propofol, nitrous oxide, and sevoflurane. AB - Anesthesia techniques in children undergoing short painful oncology procedures should allow rapid recovery without side effects. We compared the recovery characteristics of two anesthetic techniques: propofol with sevoflurane and nitrous oxide and a total IV technique using propofol and remifentanil. Twenty one children, undergoing two similar painful procedures within 2 wk were studied in a single-blind manner within patient comparison. The order of the techniques was randomized. Propofol and remifentanil involved bolus doses of both propofol 3 5 mg/kg and remifentanil 1-4 microg/kg. Propofol with sevoflurane and nitrous oxide involved propofol 3-5 mg/kg with 2%-8% sevoflurane and 70% nitrous oxide. The primary outcome variable was the time taken to achieve recovery discharge criteria; other recovery characteristics were also noted. The mean age of the children was 6.5 yr (range, 2.5-9.8 yr). Nineteen had lymphoblastic leukemia and two had lymphoma. All children had intrathecal chemotherapy and one had bone marrow aspiration. Most procedures lasted <4 min. The mean time to achieve recovery discharge criteria was appreciably shorter after propofol and remifentanil than propofol with sevoflurane and nitrous oxide by nearly 19 min (P = 0.001). All other time comparisons had similar trends and statistical differences. Seven parents expressed a preference for the propofol and remifentanil technique compared with one preferring propofol with sevoflurane and nitrous oxide. Children are apneic during the procedure and require respiratory support from an anesthesiologist. Discharge readiness from the recovery ward was achieved on average 19 min earlier after propofol with remifentanil compared with the combination of propofol, sevoflurane and nitrous oxide. Parents more often preferred propofol with remifentanil. PMID- 15781507 TI - The efficacy of fluorocarbon, surfactant, and their combination for improving acute lung injury induced by intratracheal acidified infant formula. AB - We conducted the current study to compare the efficacy of partial liquid ventilation (PLV), pulmonary surfactant (PSF), and their combination in ameliorating the acidified infant-formula-induced acute lung injury (ALI). In the Part I study, 42 rabbits receiving volume-controlled ventilation with positive end-expiratory pressure 10 cm H(2)O were randomly divided into 6 groups (groups noninjuryI, gas ventilation [GVi], PLVi, PSFi, PLVi-->PSFi, and PSFi-->PLVi). ALI was induced by intratracheal acidified infant formula (2 mL/kg, pH 1.8). Group GVi received neither PLV nor PSF therapy. Groups PLV and PSF received intratracheal fluorocarbon 15 mL/kg or surfactant 100 mg/kg, respectively, 30 min after acidified infant formula. Groups PLVi-->PSFi and PSFi-->PLVi received both treatments at 30-min intervals. In Part II, 42 rabbits (in 6 groups) undergoing pressure-controlled ventilation received the same drug therapies as in Part I. The lungs were excised to assess biochemical and histological damage 150 min after induction of ALI. In Parts I and II, PSF, fluorocarbon, and their combination attenuated lung leukosequestration and edema and superoxide production of neutrophils, consequently improving oxygenation, lung mechanics, and pathological changes. Independent of ventilation mode, PSF followed by fluorocarbon provided the most beneficial effects and fluorocarbon followed by PSF produced the least efficacy. PMID- 15781508 TI - The perioperative management of a patient with complex single ventricle physiology and pheochromocytoma. AB - Pheochromocytoma is associated with intense physiologic effects of alpha- and beta-adrenergic stimulation from catecholamine secretion. Perioperative management for these patients includes alpha-adrenergic receptor blockade, intravascular volume replacement, and, if necessary, beta-adrenergic receptor blockade. Significant perioperative changes in preload and afterload, fluid status, heart rate and rhythm, and inotropy can occur and may be contrary to anesthetic management goals for patients with certain conditions of congenital heart disease. We report the perioperative management with doxazosin of a patient with single ventricle physiology and cavo-pulmonary and aorto-pulmonary lung perfusion who presented for resection of a pheochromocytoma. PMID- 15781509 TI - Peripheral nerve blocks result in superior recovery profile compared with general anesthesia in outpatient knee arthroscopy. AB - It has been suggested that use of peripheral nerve blocks (PNBs) may have some potential benefits in the outpatient setting. There have been no studies specifically comparing PNBs performed with short-acting local anesthetics with general anesthesia (GA) in patients undergoing outpatient knee surgery. We hypothesized that a combination of lumbar plexus and sciatic blocks using a short acting local anesthetic will result in shorter time-to-discharge-home as compared with GA. Patients scheduled to undergo knee arthroscopy were randomized to receive a GA (midazolam, fentanyl, propofol, N(2)O/O(2)/desflurane via laryngeal mask airway) or lumbar plexus/sciatic block (PNBs; 2-chloroprocaine). Patients given GA also received an intraarticular injection of 20 mL 0.25% bupivacaine for postoperative pain control. Patients in the PNB group were given midazolam (up to 4 mg) and alfentanil (500-750 microg) before block placement and propofol 30-50 microg . kg(-1) . min(-1) for intraoperative sedation. Relevant perioperative times, postanesthesia care unit bypass rate, severity of pain, and incidence of complications were compared between the two groups. Fifty patients were enrolled in the study; 25 patients each received GA or PNBs. Total operating room time did not differ significantly between the 2 groups (97 +/- 37 versus 91 +/- 42 min). Seventy-two percent of patients receiving PNB met criteria enabling them to bypass Phase I postanesthesia care unit compared with only 24% of those receiving GA (P < 0.002). Time to meet criteria for discharge home (home readiness) and time to actual discharge were significantly shorter for patients given PNBs than for patients given GA (131 +/- 62 versus 205 +/- 94 and 162 +/- 71 versus 226 +/- 96, respectively). Under the conditions of our study, the combination of lumbar plexus and sciatic blocks with 2-chloroprocaine 3% was associated with a superior recovery profile compared with GA in patients having outpatient knee arthroscopy. PMID- 15781511 TI - Preventing the withdrawal response associated with rocuronium injection: a comparison of fentanyl with lidocaine. AB - We compared the efficacy of IV fentanyl with IV lidocaine as pretreatment for the prevention of withdrawal response after rocuronium injection. For this prospective, randomized, placebo-controlled, double-blind study we recruited 90 patients aged between 18 and 65 yr, ASA physical status I or II, who had undergone elective surgery requiring general anesthesia and positive pressure ventilation. Patients were randomly allocated to 1 of 3 groups: group F received 2 mL IV fentanyl 50 microg/mL (100 microg), group L received 2 mL of preservative free lidocaine 2% (40 mg), and group P (placebo) received 2 mL of normal saline. The incidence of withdrawal response after rocuronium was 57%, 30%, and 7% in the placebo, lidocaine, and fentanyl groups, respectively. We found a significant reduction in incidence of withdrawal response in both the fentanyl and lidocaine groups when compared with the placebo group (P < 0.05), with the fentanyl group being most effective (P < 0.05). In conclusion, both fentanyl and lidocaine are effective clinical treatments to alleviate the withdrawal response associated with rocuronium injection, with fentanyl being more effective. PMID- 15781512 TI - The role of adrenergic and cholinergic transmission in volatile anesthetic induced pain enhancement. AB - Volatile anesthetic drugs have a biphasic effect on pain transmission. At very small concentrations they enhance pain sensitivity whereas at larger subanesthetic concentrations they have an analgesic effect. Previous work has suggested that nicotinic inhibition could mediate the pronociceptive action of isoflurane. Furthermore, activation of nicotinic receptors facilitates the release of norepinephrine in the spinal cord. We hypothesize that nicotinic modulation of norepinephrine release in the spinal cord mediates isoflurane's pronociceptive action. We used hindpaw withdrawal latency as a measure of pain sensitivity after inhibition of adrenergic activity or treatment with nicotine in mice. Isoflurane's effect on pain is separable by concentration. The 50% effective concentration for pain enhancement is 0.16% isoflurane whereas the 50% effective concentration for the antinociceptive action of isoflurane is 0.8%. Depletion of systemic norepinephrine with the neurotoxin DSP-4 caused a reduction in baseline withdrawal latencies and prevented isoflurane pronociception. Baseline latency was also reduced by intrathecal yohimbine. After treatment with yohimbine, isoflurane had no additional pronociceptive effect. Nicotine administered through intracerebroventricular injection increased baseline latency but did not prevent isoflurane pronociception. Conversely, intrathecal applications of nicotine caused a slight reduction in baseline latency and prevented isoflurane's pronociceptive effect. We conclude that spinal noradrenergic transmission seems to be necessary for isoflurane pronociception to occur. Isoflurane may act by inhibiting tonically active nicotinic receptors that modulate the release of norepinephrine in the spinal cord. PMID- 15781513 TI - The effect of mild hypothermia on plasma fentanyl concentration and biotransformation in juvenile pigs. AB - Therapeutic hypothermia may alter the required dosage of analgesics and sedatives, but no data are available on the effects of mild hypothermia on plasma fentanyl concentration during continuous, long-term administration. We therefore assessed in a porcine model the effect of prolonged hypothermia on plasma fentanyl concentration during 33 h of continuous fentanyl administration. Seven female piglets (weight: 11.8 +/- 1.1 kg) were anesthetized by IV fentanyl (15 microg . kg(-1) . h(-1)) and midazolam (1.0 mg . kg(-1) . h(-1)). After preparation and stabilization (12 h), the animals were cooled to a core temperature of 31.6 degrees +/- 0.2 degrees C for 6 h and were then rewarmed and kept normothermic at 37.7 degrees +/- 0.3 degrees C for 6 more hours. Plasma fentanyl concentrations were measured by radioimmunoassay, cardiac index by thermodilution, and blood flows of the kidney, spleen, pancreas, stomach, gut, and hepatic artery by a colored microspheres technique. Furthermore, in an additional 4 pigs, temperature dependency of hepatic microsomal cytochrome P450 3A4 (CYP3A4) was determined in vitro by ethylmorphine N-demethylation. Plasma fentanyl concentration increased by 25% +/- 11% (P < 0.05) during hypothermia and remained increased for at least 6 h after rewarming. Hypothermia reduced the cardiac index (41% +/- 15%, P < 0.05), as well as all organ blood flows except the hepatic artery. A strong temperature dependency of CYP3A4 was found (P < 0.01). Mild hypothermia induced a distribution and/or elimination-dependent increase in plasma fentanyl concentration which remained increased for several hours after rewarming. Consequently, a prolonged increase of the plasma fentanyl concentration should be anticipated for appropriate control of the analgesia/sedatives during and early after therapeutic hypothermia. PMID- 15781514 TI - Solubility of volatile anesthetics in bovine white matter, cortical gray matter, thalamus, hippocampus, and hypothalamic area. AB - Although known for whole brain, values are lacking for solubilities of modern volatile anesthetics in specific brain regions. Some regions should differ from others (e.g., gray matter versus white matter) because they differ in lipid content and because potent inhaled anesthetics are lipophilic. In the present report, we examined this issue in bovine brain, finding that white matter/gas partition coefficients are 1.6 (desflurane) to 2.4 (halothane) times larger than gray matter/gas partition coefficients, with values for isoflurane and sevoflurane lying between these at 1.9. Values for thalamus/gas, hypothalamic area/gas, and hippocampal/gas partition coefficients lie between those for gray and white matter. These data may be useful in defining the parts of the brain involved with return to consciousness during recovery from anesthesia. PMID- 15781515 TI - The in vitro effect of desflurane preconditioning on endothelial adhesion molecules and mRNA expression. AB - Lower expression of intercellular adhesion molecule-1 (ICAM-1), vascular adhesion molecule-1 (VCAM-1), and E-selectin may be responsible for attenuated ischemic reperfusion neutrophil adhesion to vascular endothelium. Desflurane reduces ischemia-reperfusion injury. Therefore, we assessed whether desflurane affects the protein expression of ICAM-1 and E-selectin and mRNA expression of ICAM-1 and VCAM-1 of human umbilical venous endothelial cells (HUVEC) stimulated with tumor necrosis factor-alpha (TNF-alpha). HUVEC were preconditioned for 60 min with 1 minimum alveolar concentration desflurane before stimulating with TNF-alpha. Protein expression of adhesion molecules ICAM-1 and E-selectin of HUVEC were evaluated via immunocytochemical techniques combined with image cytometry. ICAM-1 and VCAM-1 mRNA expression of HUVEC were determined via reverse transcription polymerase chain reaction. Desflurane not only reduced the protein expression of ICAM-1 and E-selectin but also ICAM-1 and VCAM-1 mRNA expression of the HUVEC. The adhesion rate of neutrophils with desflurane-treated HUVEC was slower. The decreased neutrophil adhesion on the desflurane-treated HUVEC correlated well with the decrease in adhesion molecule expression. These results show that desflurane affects the expression of adhesion molecules involved in the multistep process of neutrophil recruitment. Desflurane related ischemia-reperfusion injury reduction correlates well with expression inhibition of ICAM-1, VCAM-1, and E selectin that mediates neutrophil rotation and firm adhesion on the vascular endothelium. PMID- 15781516 TI - A comparative evaluation of inhaled halothane, isoflurane, and sevoflurane during acute normovolemic hemodilution in dogs. AB - The hemodynamic response to acute normovolemic hemodilution (ANH) can be affected by the anesthetics used. We randomized 18 mongrel dogs to undergo ANH with 3 different inhaled anesthetics: halothane, isoflurane, or sevoflurane. Hemodynamics, oxygen transport, and gastric pH were measured before blood withdrawal, at the end of hemodilution, and 30 and 60 min after the end of hemodilution. The baseline measurements of all hemodynamic variables were similar among groups, with the exception of heart rate, which was more rapid in the sevoflurane group. Thirty minutes after hemodilution, the cardiac index increased 88%, 86%, and 157% in the halothane, isoflurane, and sevoflurane groups, respectively, whereas arterial-venous oxygen differences and oxygen consumption were larger in the halothane group compared with the isoflurane and sevoflurane groups. Gastric pH obtained by tonometry did not change and was not different among groups. Because the hemodynamic response to ANH was not blunted, all three anesthetics may be safely used for the maintenance of anesthesia. PMID- 15781517 TI - Illustrations of inhaled anesthetic uptake, including intertissue diffusion to and from fat. AB - Although several mathematical and computer simulations of inhaled anesthetic pharmacokinetics have been devised, their complexity sometimes limits an intuitive appreciation of the interactions produced by the determinants of kinetics. In this essay, we illustrate the factors that govern inhaled anesthetic pharmacokinetics with drawings that consider delivery of anesthetic by ventilation to the lungs and dispersion of the anesthetic to tissue depots by the circulation. The illustrations incorporate the effects of both blood flow and blood solubility as determinants of the extent of dispersion. They incorporate tissue volume and solubility as determinants of the capacity of the tissue depots. Capacity to hold (take up) anesthetic is depicted by areas representing specific tissues, and the extent of anesthetic movement is depicted by the length and breadth of arrows to and from the areas depicting capacity. The illustrations incorporate increasingly important elements to kinetics, such as obesity. Obesity increases the depots available for storage of anesthetic, including anesthetic that reaches fat by intertissue diffusion. Such anesthetic returns to the circulation to delay recovery in healthy and obese patients, particularly with more soluble anesthetics. However, the increased anesthetic in fat occurs at a lower partial pressure and thus might not influence emergence materially. We hope that these illustrations will allow anesthesia practitioners to appreciate the interactions of the factors that govern inhaled anesthetic pharmacokinetics. PMID- 15781518 TI - RGS proteins: new players in the field of opioid signaling and tolerance mechanisms. AB - In this article we review recent advances in our understanding of the crucial role of the Regulator of G protein Signaling (RGS) proteins in opioid signaling mechanisms and opioid tolerance development. Opioids exert their physiologic effects via complex G protein-coupled receptor-signaling mechanisms, and RGS proteins are now known to tightly regulate the G protein signaling cycle. RGS proteins contain GTPase-accelerating protein activity within their characteristic RGS domain and various other receptor signaling-related properties of their other functional domains. There have been more than 20 RGS proteins reported in the literature, and multiple RGS proteins have been shown to negatively regulate G protein-mediated opioid signaling, facilitate opioid receptor desensitization and internalization, and affect the rate at which opioid tolerance develops. Using RGS proteins as targets for future drug therapy aimed at modulating opioid effectiveness in both acute and chronic pain settings may be an important advance in the treatment of pain. PMID- 15781519 TI - Twelve-lead high-frequency QRS electrocardiography during anesthesia in healthy subjects. AB - Analysis of the high-frequency (HF) components of the QRS complex has been shown to be a more sensitive indicator of myocardial ischemia and infarction than conventional ST segment analysis in settings outside of the operating room. In this study, we documented the effect of general anesthesia on HF QRS analysis in healthy patients as the first step in determining the potential of this technique for monitoring anesthetized patients. HF QRS electrocardiograms (ECGs) were obtained from all 12 ECG leads in 30 healthy subjects before and after the induction of anesthesia. When compared with preinduction values, there were significant postinduction changes in multiple variables of the HF QRS in many leads studied that were within previously described normal limits. Additional study is needed to understand the potential of this monitoring technique for enhancing detection of myocardial ischemia in the anesthetized population. PMID- 15781520 TI - The impact of carrier flow rate and infusion set dead-volume on the dynamics of intravenous drug delivery. AB - The dynamics of IV drug delivery resulting from drug infusions connected to main line crystalloid carriers can be complex and depend on infusion set dead-volume, drug flow rate, and carrier flow rate. While the concept of dead-volume is intuitive, a lack of appreciation of the interaction with the carrier and drug flow rates can lead to unintended clinical effects resulting from large variations in the delivery rate of potent drugs. We derived mathematical models to quantify these interactions. Experimental simulation with methylene blue infusions tested these predictions. The models predict a lag in response time to changes in carrier or drug flow, which is proportional to the dead-volume and inversely related to the total flow rate. Increasing the carrier rate provides an acute drug bolus. Temporary reduction or cessation of carrier flow decreases the rate of drug delivery, potentially for prolonged periods. Furthermore, a drug bolus results from restoration of the carrier flow. The method of connecting an infusion to a carrier and the use history affects the dynamics of drug delivery. Thus, although complex, the impact of infusion set architecture and changes in carrier and drug flow rates are predictable. These quantitative studies may help optimize the safe use of IV drug infusion systems. PMID- 15781521 TI - Non-zero basal oxygen flow a hazard to anesthesia breathing circuit leak test. AB - The non-zero basal flow (BF) of oxygen in anesthesia machines has been set to supply the basal metabolic requirement of oxygen. However, there is no scientific evidence of its necessity. In this study we sought to clarify whether non-zero BF affects leak detection during preanesthetic inspections. Twenty-five participants performed leak tests on anesthesia machines to detect breathing circuit leaks. Artificial leak-producing devices were used to create leaks from 0 to 1.0 L/min. The investigator randomly chose the leak device and connected it into the breathing circuit. Participants, blinded as to the presence or the type of leak producing device, then tested the breathing circuit for leaks. The conventional breathing system leak test was performed with and without BF. The results of leak detection in each leak procedure were analyzed statistically. The leak detection rate of leak test with BF was less than without BF (P < 0.01). We demonstrated that non-zero BF of oxygen decreases the leak detection rate and is an obstacle for leak detection, especially for small leaks. Therefore, we recommend that breathing circuit leak tests should be performed in the absence of BF of oxygen. PMID- 15781522 TI - A comparison of bispectral index and entropy, or how to misinterpret both. AB - Consciousness monitoring has become increasingly popular in general anesthesia cases, and a new technology has recently been introduced with potential advantages over the other available products. In this case report, we discuss a patient who was monitored simultaneously with Bispectral Index and Entropy and evaluate the differences between the two. More importantly, we emphasize the importance of vigilance when using new technologies and discuss the potential impact of lack of vigilance on patient outcome. PMID- 15781523 TI - Surgical field fire during a repair of bronchoesophageal fistula. AB - Most surgical fires involve the airway but they can also occur in the surgical field. Herein, we report an intraoperative fire in the surgical field during repair of a bronchoesophageal fistula. During the portion of the surgery after the fistula was divided and the bronchus was open to atmosphere, continuous positive airway pressure was applied to the nondependent lung, and in conjunction with the use of electrocautery and dry sponges in the field, resulted in a fire. Anesthesia for thoracic surgery carries unique risks of fire because these patients frequently require large oxygen concentrations, special interventions for improving oxygenation, and have variable degrees of airway disruption. This report highlights unique safety concerns during anesthesia for thoracic surgery, and addresses more general safety issues relating to fire risk in all surgical patients. PMID- 15781524 TI - A comparison of Depodur, a novel, single-dose extended-release epidural morphine, with standard epidural morphine for pain relief after lower abdominal surgery. AB - In this randomized, controlled, dose-ranging study, we evaluated the analgesic efficacy of a novel single-dose extended-release epidural morphine (Depodur) in patients undergoing lower abdominal surgery. Five-hundred-forty-one patients were randomly assigned to one of six epidural treatments administered approximately 30 min before surgery. The 6 treatments were 5 mg of standard epidural morphine sulfate (MS) (active comparator); 5 mg of single-dose extended-release epidural morphine (EREM) (dose control); and 10, 15, 20, and 25 mg of single-dose EREM. The main study objective was to assess the efficacy of single-dose EREM 10, 15, 20, or 25 mg versus single-dose EREM 5 mg for the management of postoperative pain. This was done by plotting a linear dose-response relationship to assess postoperative IV patient-controlled analgesia (PCA) fentanyl consumption for breakthrough pain for 48 h after surgery. Secondary safety and efficacy analyses compared the 10-, 15-, 20-, and 25-mg single-dose EREM groups with the 5-mg single-dose EREM group and compared each single-dose EREM group with 5 mg of MS. As shown by the dose-response relationship, there was a dose-related reduction in the use of postoperative IV fentanyl through 48 h (estimated slope, -22.2; P = 0.0002). Patients treated with 10, 20, and 25 mg of single-dose EREM used significantly less IV fentanyl (mean +/- sd: 995 +/- 987 microg, P = 0.0446; 972 +/- 982 microg, P = 0.0221; and 683 +/- 620 microg, P < 0.0001, respectively) through 48 h after surgery compared with the 5-mg single-dose EREM group (1218 +/ 894 microg). At 48 h postdose, significantly more single-dose EREM patients (13%) than MS patients (2%) had required no IV fentanyl (P < 0.01). Although all treatment groups had access to PCA fentanyl and there was more frequent PCA fentanyl use in the MS group, patients in the single-dose EREM 15, 20, and 25 mg groups reported significantly lower pain-intensity scores and greater satisfaction with their pain relief. Overall, single-dose EREM was well tolerated, with 97% of adverse events rated as mild to moderate. As expected, the adverse events reported were consistent with those of other epidural opioids (i.e., nausea, vomiting, pruritus, and hypotension). In conclusion, this controlled study demonstrated that single-dose EREM can provide up to 48 h of postoperative analgesia, but supplementation for breakthrough pain is still required in most patients. Within the context of this study, the side effect profile of single-dose EREM was acceptable and predictable. PMID- 15781525 TI - Intrathecal neostigmine prevents intrathecal clonidine from attenuating hypercapnic cerebral vasodilation in rabbits. AB - We previously demonstrated that lumbar intrathecal alpha(2) agonists attenuate hypercapnia-induced cerebral vasodilation. The combination of intrathecal clonidine and neostigmine is being investigated as pain therapy. The effects of their combination on cerebrovascular reactivity are unknown. We allocated rabbits anesthetized with pentobarbital to two groups: (a) clonidine (normal saline followed 30 min later by clonidine 2 microg/kg, both into the lumbar intrathecal space; n = 6), and (b) neostigmine-pretreatment (neostigmine 2 microg/kg followed 30 min later by clonidine 2 microg/kg, both into the lumbar intrathecal space; n = 6). We then evaluated the hypercapnia-induced changes in pial arteriolar diameter in these two groups using the closed cranial window preparation. The pial arteriolar dilator response to hypercapnia was significantly attenuated in the clonidine group (14% +/- 4%, 4% +/- 4%, 6% +/- 6%, and 5% +/- 7% for before and 30, 60, and 90 min, respectively). Neither normal saline nor neostigmine alone induced any change in the cerebral reactivity to hypercapnia. Pretreatment with neostigmine completely prevented the clonidine-induced attenuation of the hypercapnic cerebral vasodilation attenuated by intrathecal clonidine (16% +/- 7%, 15% +/- 6%, 12% +/- 6%, and 16% +/- 8%, respectively). PMID- 15781526 TI - Bupivacaine's action on the carrageenan-induced inflammatory response in mice: cytokine production by leukocytes after ex-vivo stimulation. AB - We aimed to study the effect of bupivacaine on the systemic response elicited by intraplantar injection of carrageenan. To that purpose, we studied the effects of carrageenan, bupivacaine, or both on the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-10 by whole blood cultured in the presence of lipopolysaccharide (LPS) and of heat-killed Staphylococcus Aureus Cowan (SAC). Mice received a hindpaw injection of carrageenan with or without encapsulated IM bupivacaine given contralaterally. Whole blood was sampled 15 h later and cultured for 24 h with LPS or SAC. The amounts of TNF-alpha, IL-1beta, and IL-10 in the supernatants were measured. In the presence of LPS or SAC, proinflammatory cytokine (TNF-alpha and IL-1beta) production was increased after carrageenan. Bupivacaine prevented this inflammatory response: 992 +/- 102 versus 2146 +/- 338 versus 919 +/- 116 pg/mL for TNF-alpha (bupivacaine + carrageenan versus carrageenan versus control after LPS stimulation). This effect of bupivacaine was less after SAC stimulation. Moreover, IL-10 was not involved in the inhibition of proinflammatory cytokine production observed after treatment by bupivacaine alone. These experiments show that carrageenan-induced hindpaw inflammation modifies the blood cell reactivity to LPS and SAC and that bupivacaine regulates the systemic response elicited by carrageenan. Furthermore, IL-10 does not seem to be a factor of the antiinflammatory response induced by bupivacaine. The precise mechanism underlying this effect of bupivacaine remains to be clarified. PMID- 15781528 TI - Perioperative fluid management and clinical outcomes in adults. AB - The administration of IV fluid to avoid dehydration, maintain an effective circulating volume, and prevent inadequate tissue perfusion should be considered, along with the maintenance of sleep, pain relief, and muscular relaxation, a core element of the perioperative practice of anesthesia. Knowledge of the effects of different fluids has increased in recent years, and the choice of fluid type in a variety of clinical situations can now be rationally guided by an understanding of the physicochemical and biological properties of the various crystalloid and colloid solutions available. However, there are few useful clinical outcome data to guide this decision. Deciding how much fluid to give has historically been more controversial than choosing which fluid to use. A number of clinical studies support the notion that an approach based on administering fluids to achieve maximal left ventricular stroke volume (while avoiding excess fluid administration and consequent impairment of left ventricular performance) may improve outcomes. In this article, we review the available fluid types and strategies of fluid administration and discuss their relationship to clinical outcomes in adults. PMID- 15781527 TI - Spinal opioid receptor like1 receptor agonist, but not N-methyl-D-aspartic acid antagonist, reverses the secondary mechanical allodynia induced by intradermal injection of capsaicin in rats. AB - Secondary mechanical allodynia induced by intradermal injection of capsaicin has been widely used to search for the underlying mechanisms of tissue injury induced mechanical allodynia. However, the capsaicin concentration dependency of the development of secondary mechanical allodynia and the underlying mechanisms of development and maintenance of capsaicin-induced mechanical allodynia are not fully understood. In the present study, we clarify the capsaicin concentration dependency for development and maintenance of secondary mechanical allodynia and the role of spinal opioid receptor like1 (ORL1) receptor and N-methyl-D-aspartate receptor in the development and maintenance of secondary mechanical allodynia induced by an intradermal capsaicin injection. Capsaicin 50 microL of 0.03% induced the most intense secondary mechanical allodynia. Intrathecal injection of nociceptin, an ORL1 receptor agonist, attenuated the maintenance of secondary mechanical allodynia but had no effect on the development of secondary mechanical allodynia. An intrathecal injection of MK801, an N-methyl-D-aspartate receptor antagonist, had no effect on the development and maintenance of secondary mechanical allodynia. These findings suggest that spinal ORL1 receptor should be the target of study for the treatment of secondary mechanical allodynia induced by tissue injury. PMID- 15781529 TI - The effect of nitroglycerin on microvascular perfusion and oxygenation during gastric tube reconstruction. AB - Esophagectomy followed by gastric tube reconstruction is the surgical treatment of choice for patients with esophageal cancer. Complications of the cervical anastomosis are associated with impaired microvascular blood flow (MBF) and ischemia in the gastric fundus. The aim of the present study was to differentiate whether the decrease in MBF is a result of arterial insufficiency or of venous congestion. To do this we assessed MBF, microvascular hemoglobin oxygen saturation (muHbSo(2)), and microvascular hemoglobin concentration (muHbcon) simultaneously during different stages of gastric tube reconstruction. In 14 patients, MBF was determined with laser Doppler flowmetry, and muHbSo(2) and muHbcon were determined with reflectance spectro- photometry. After completion of the anastomosis, nitroglycerin was applied at the fundus. Although MBF did not change significantly in the pylorus, MBF decreased progressively during surgery in the fundus from 210 +/- 18 Arbitrary Units at baseline (normal stomach) to 52 +/- 9 Arbitrary Units after completion of reconstruction (mean +/- sem; P < 0.05). There was no change in muHbSo(2) and muHbcon during the reconstruction. After application of nitroglycerin, MBF doubled. We conclude that MBF decreases during gastric tube reconstruction but that muHbSo(2) and muHbcon do not. This decrease might be the result of venous congestion, which can partly be counteracted by application of nitroglycerin. PMID- 15781530 TI - The effect of positive end-expiratory pressure level on peak expiratory flow during manual hyperinflation. AB - Including positive end-expiratory pressure (PEEP) in the manual resuscitation bag (MRB) may render manual hyperinflation (MHI) ineffective as a secretion maneuver technique in mechanically ventilated patients. In this study we aimed to determine the effect of increased PEEP or decreased compliance on peak expiratory flow rate (PEF) during MHI. A blinded, randomized study was performed on a lung simulator by 10 physiotherapists experienced in MHI and intensive care practice. PEEP levels of 0-15 cm H(2)O, compliance levels of 0.05 and 0.02 L/cm H(2)O, and MRB type were randomized. The Mapleson-C MRB generated significantly higher PEF (P < 0.01, d = 2.72) when compared with the Laerdal MRB for all levels of PEEP. In normal compliance (0.05 L/cm H(2)O) there was a significant decrease in PEF (P < 0.01, d = 1.45) for a PEEP more than 10 cm H(2)O in the Mapleson-C circuit. The Laerdal MRB at PEEP levels of more than 10 cm H(2)O did not generate a PEF that is theoretically capable of producing two-phase gas-liquid flow and, consequently, mobilizing pulmonary secretions. If MHI is indicated as a result of mucous plugging, the Mapleson-C MRB may be the most effective method of secretion mobilization. PMID- 15781531 TI - A comparison of the efficacy of heparinized and nonheparinized solutions for maintenance of perioperative radial arterial catheter patency and subsequent occlusion. AB - In a randomized, double-blind, controlled study, we compared heparinized and nonheparinized infusions for the maintenance of perioperative arterial catheter patency and the incidence of subsequent radial arterial occlusion. Two-hundred patients were randomized into 2 groups to receive heparinized (group H, n = 100) or nonheparinized (group S, n = 100) flush solutions. Radial and ulnar blood flows were assessed using Doppler probe and pulse oximetry before, just after, and 24 h after decannulation by the same investigator. The cannulation site was examined for complications such as hematoma, nerve injury, and infection. The mean duration of cannulations was 378 +/- 159.0 min in group H and 332 +/- 154.6 min in group S. The mean number of corrective interventions caused by dampening of the pressure wave (mean number of positional changes [group S, 1.5 +/- 2.0; group H, 1.4 +/- 3.8] and mean number of manual flushes [group S, 1.3 +/- 1.7; group H, 1.2 +/- 1.2]) was not significantly different in both groups. After decannulation, partial or total occlusion developed in 20 group H patients and 16 group S patients (not significant). The incidence of occlusion was correlated to the presence of hematoma at the puncture site after decannulation (P = 0.013), long duration of cannulation (P = 0.04), and age <65 yr (P = 0.009). In conclusion, there is no significant difference between heparinized and nonheparinized flush solutions for the maintenance of perioperative radial artery catheter patency. PMID- 15781532 TI - Perfluorohexane vapor has only minor effects on spatial pulmonary blood flow distribution in isolated rabbit lungs. AB - We tested the hypothesis that administration of perfluorohexane (PFH) vapor does not significantly affect the relative pulmonary blood flow (Qrel) distribution in isolated rabbit lungs. Fourteen isolated rabbit lungs were perfused with a Krebs Henseleit buffer solution (flow 150 mL/min). Pulmonary afterload was set to 3 mm Hg. The lungs were ventilated with 4% CO(2) in room air using a small animal ventilator (respiratory rate, 30 breaths/min; tidal volume, 12 mL/kg body weight; positive end-expiratory pressure, 2 cm H(2)O). After a steady-state period, 18 vol. % of PFH vapor was administered to 9 lungs for 30 min. In a second set of experiments five lungs served as controls. Change in (Qrel) distribution was assessed using fluorescent-labeled microspheres. The unpaired Student's t-test was used to compare variables between groups. The paired Student's t-test, the one-sample Student's t-test, the Anderson-Hauck test of equivalence, and Pearson correlation were used to analyze changes within groups. The mean correlation coefficients of (Qrel) were 0.564 +/- 0.182 for the PFH group and 0.502 +/- 0.295 for the control group, respectively. No significant changes in (rel) distribution over time and between groups were found. However, in the PFH group a tendency towards redistribution of (Qrel) to more ventral lung areas was noted. Our results suggest that PFH vapor has no significant effects on redistribution of (Qrel) in isolated rabbit lungs. PMID- 15781533 TI - Hyperglycemia in patients administered dexamethasone for craniotomy. AB - Hyperglycemia should be avoided during neurosurgery in order to decrease the risk of neurological injury. Dexamethasone has been associated with increased blood glucose during surgery. In this prospective, nonrandomized study, we documented the blood glucose concentration changes for 12 h in 34 nondiabetic patients undergoing craniotomy and compared patients who received intraoperative dexamethasone (10 mg IV on induction and 4 mg IV 6 h later), with or without preoperative dexamethasone, with patients who did not receive dexamethasone. Blood glucose concentrations increased from the preinduction value in all groups. Patients not taking dexamethasone before surgery, but who were given it intra- and postoperatively, had the largest peak blood glucose concentrations (11.0 +/- 2.0 mmol/L, mean +/- sd; P < 0.01) compared with patients who received no dexamethasone (7.8 +/- 2.1 mmol/L) or those who had been taking dexamethasone before surgery and continued it during surgery (8.5 +/- 1.2 mmol/L). The peak blood glucose concentrations in this group occurred 9 +/- 2 h after the induction of anesthesia. We recommend that the blood glucose concentration be monitored for at least 12 h in nondiabetic patients having neurosurgery who are newly administered dexamethasone. PMID- 15781534 TI - Intrathecal ketorolac pretreatment reduced spinal cord ischemic injury in rats. AB - Paraplegia caused by spinal cord ischemic injury remains a potential complication of surgical repair of thoracoabdominal aortic aneurysms. Studies suggest that cyclooxygenase (COX) contributes to ischemic neuronal damage and that COX inhibitors may reduce injury. In this study, we examined whether intrathecal pretreatment with ketorolac, a nonselective COX inhibitor, had a protective effect against ischemic spinal cord injury in rats. Rats were randomized to receive either intrathecal normal saline, ketorolac 30 microg, or ketorolac 60 microg (n = 6 rats per group) 1 h before spinal cord ischemia (intraaortic balloon occlusion combined with proximal arterial hypotension for 11 min). Another 6 rats served as the sham-operated controls. Ischemic injury was assessed by hindlimb motor function and by histopathological changes in the lumbar spinal cord at 24 h after the ischemic insult. The other 20 rats (n = 10 per group) were used in the second experiments to evaluate the safety of this drug. Survival of rats was recorded 28 days after reperfusion. Intrathecal pretreatment with 60 microg of ketorolac significantly reduced neuronal death and improved hindlimb motor function, and the long-term survival was similar to that in the control group. The results suggest that intrathecal ketorolac may be of therapeutic potential for preventing spinal cord ischemic injury during thoracoabdominal aortic surgery. PMID- 15781535 TI - Pial arterial response to topical verapamil in acute closed cranial windows in rabbits. AB - We have previously observed that intraarterial verapamil increases cerebral blood flow, whereas nitric oxide donors, such as nitroglycerin, do not. Clinically, both verapamil and nitroglycerin dilate large cerebral arteries. Therefore, we hypothesized that topical verapamil would dilate both the large proximal and the small distal cerebral arteries, whereas nitroglycerin would preferentially dilate only the large proximal arteries. We tested our hypothesis in acute cranial windows implanted in 10 New Zealand White rabbits. After predrug measurements, we superfused 4 increasing concentrations of verapamil or nitroglycerin (10(-8), 10( 6), 10(-4), and 10(-3) M) in the cranial windows for 5 min each. The maximum increase in diameter was expressed as a percentage change from predrug diameters. There was a 30-min period of rest between the two drug challenges. Topical verapamil increased the arterial diameter of the larger proximal arterioles (>60 microm) by 32% +/- 18% and that of the smaller distal arterioles (<60 microm) by 62% +/- 42%. A modest increase in arterial diameters of 11% +/- 11% was observed after topical nitroglycerin that was significant only for the large-proximal arterioles. Within the 10(-8) to 10(-3) M range, topical verapamil, compared with nitroglycerin, proved to be a more potent cerebral vasodilator and had a more robust vasodilator effect on the distal small pial arteries. PMID- 15781536 TI - Delayed cardiogenic shock and acute lung injury after aneurysmal subarachnoid hemorrhage. AB - Both cardiac and lung injury after aneurysmal subarachnoid hemorrhage has been attributed to an adrenergic surge. Cardiogenic shock is very uncommon. We describe a 55-yr-old woman with a delayed cardiogenic shock emerging within hours after aneurysmal rupture. Cardiac damage was documented by increased serum troponin T, CPK-mb fraction, and severe wall motion abnormality, which included an akinetic apex on echocardiography (ejection fraction of 33%). Her coronary angiogram was normal. Decreased cardiac index, increased systemic and pulmonary vascular resistance indices, and persistent oxygen desaturation despite improving ventricular contractility documented both cardiac and pulmonary injury. After treatment with dobutamine and milrinone all manifestations resolved. PMID- 15781537 TI - Single-dose, sustained-release epidural morphine in the management of postoperative pain after elective cesarean delivery: results of a multicenter randomized controlled study. AB - In this multicenter, randomized, controlled study, we compared the analgesic efficacy and safety profile of a new single-dose extended-release epidural morphine (EREM) formulation (DepoDur) with that of epidural morphine sulfate for the management of postoperative pain for up to 48 h after elective cesarean delivery. ASA physical status I or II parturients (n = 75) were anesthetized with a combined spinal/epidural technique. Parturients received intrathecal bupivacaine 12-15 mg and fentanyl 10 mug for spinal anesthesia and a single epidural injection of either 5 mg of standard (conventional preservative-free) morphine or 5, 10, or 15 mg of extended-release morphine after cord clamping for postoperative pain control. Single-dose EREM 10 and 15 mg groups significantly decreased total supplemental opioid medication use and improved functional ability scores for 48 h after surgery compared with those receiving 5 mg of standard morphine. Visual analog scale pain scores at rest and with activity at 24 to 48 h after dosing were significantly better in the 10- and 15-mg single dose EREM groups versus the standard morphine group. There were no significant differences between the two 5 mg (single-dose EREM and standard morphine) groups. Single-dose EREM was well tolerated, and most adverse events were mild to moderate in severity. Single-dose EREM is a potentially beneficial epidural analgesic for the management of post-cesarean delivery pain and has particular advantages over standard morphine for the period from 24 to 48 h after surgery. PMID- 15781538 TI - The effect of posture and baricity on the spread of intrathecal bupivacaine for elective cesarean delivery. AB - Posture and baricity during induction of spinal anesthesia with intrathecal drugs are believed to be important in determining spread within the cerebrospinal fluid. In this double-blind prospective study, 150 patients undergoing elective cesarean delivery were randomized to receive a hyperbaric, isobaric, or hypobaric intrathecal solution of 10 mg bupivacaine during spinal anesthesia induced in either the sitting or right lateral position. After an intrathecal injection using a combined-spinal technique patients were placed in the supine wedged position. We determined the densities of the three intrathecal solutions from a previously validated formula and measured using a DMA-450 density meter. Data collection included sensory level, motor block, episodes of hypotension, and ephedrine use. Statistical analysis included analysis of variance and Cuzick's trend. In the lateral position, baricity had no effect on the spread of sensory levels for bupivacaine compared to the sitting position, where there was a statistically significant difference in spread with the hypobaric solution producing higher levels of analgesia than the hyperbaric solution (P = 0.002). However, the overall differences in maximal spread only differed by one dermatome, with the hyperbaric solution achieving a median maximum sensory level to T3 compared with T2 for the isobaric and hypobaric solutions. Motor block was significantly (P = 0.029) reduced with increasing baricity and this trend was significant (P = 0.033) for the lateral position only. Hypotension incidence and ephedrine use increased with decreasing baricity (P = 0.003 and 0.004 respectively), with the hypobaric sitting group having the most frequent incidence of hypotension (76%) as well as cervical blocks (24%; P = 0.032). PMID- 15781539 TI - Interscalene brachial plexus block with bupivacaine and ropivacaine in patients with chronic renal failure: diaphragmatic excursion and pulmonary function changes. AB - In this randomized, double-blind study, we compared the anesthetic characteristics and pulmonary function changes of 0.33% bupivacaine and 0.33% ropivacaine used for interscalene brachial plexus (IBP) anesthesia in patients with chronic renal failure. Forty-two patients undergoing IBP anesthesia for creation of arteriovenous fistulas were randomly allocated to receive either 30 mL of 0.33% bupivacaine (Group B) or 0.33% ropivacaine (Group R). Block onset time, diaphragmatic excursion (ultrasonographic evaluation), and free plasma concentrations of bupivacaine and ropivacaine were evaluated. Negative motion or immobility of the ipsilateral hemidiaphragm and a decrease of >10 mm in positive motion were defined as diaphragmatic paresis. The pulmonary function variables were measured by bedside spirometry equipment. Seven patients needed supplemental local anesthetic, one with total spinal block; these patients were excluded from the study. The success rate was 80.9%. Block quality was similar in the two groups. Ipsilateral hemidiaphragmatic excursion was decreased in both groups compared with baseline values (P < 0.05). Diaphragmatic paresis was identified in 10 of 16 patients and 8 of 18 patients in Groups B and R, respectively (P > 0.05). Pulmonary function significantly decreased from baseline in both groups (forced vital capacity (FVC) 30%, forced expiratory volume at 1 second (FEV(1)) 32%, and peak expiratory flow (PEF) 31% in Group B and FVC 17%, FEV(1) 17%, and PEF 5% in Group R) (P < 0.001). The decreases in Group B were larger than those in Group R (P < 0.05). Three patients in Group B and one in Group R had mild respiratory problems (P > 0.05). Concentrations of bupivacaine and ropivacaine were below toxic levels rather than "normal range." We conclude that pulmonary function decreased more after IBP with 0.33% bupivacaine than with 0.33% ropivacaine. PMID- 15781540 TI - Clonidine added to a continuous interscalene ropivacaine perineural infusion to improve postoperative analgesia: a randomized, double-blind, controlled study. AB - Although clonidine has been shown to increase the duration of local anesthetic action and prolong postoperative analgesia when included in single-injection nerve blocks, the only controlled investigation of the efficacy of this practice to improve analgesia for continuous perineural local anesthetic infusion failed to discern any clinically relevant benefits. For this study, we used a larger dose of clonidine in an attempt to improve analgesia. Patients (n = 20) undergoing moderately painful orthopedic surgery of the shoulder received an interscalene brachial plexus block (40 mL of mepivacaine 1.5%, epinephrine 2.5 microg/mL, and clonidine 50 microg) and a perineural catheter before surgery. After surgery, ropivacaine 0.2% or ropivacaine 0.2% plus clonidine 2 microg/mL was delivered via the catheter for 3 days (basal rate, 5 mL/h; patient-controlled bolus, 5 mL; lockout, 1 h). Investigators and patients were blind to random group assignment. The primary outcome variable was designated as the most intense pain during the day after surgery. Secondary end-points included additional pain scores, patient-controlled bolus doses, oral analgesic use, sleep quality, and catheter- or infusion-related complications. There were no statistically significant differences between groups for any of the variables investigated. We conclude that adding clonidine 2 microg/mL to a ropivacaine interscalene perineural infusion does not decrease breakthrough pain intensity the day after surgery. For the additional end-points, our negative findings are only suggestive of a lack of effect and require further study for verification. PMID- 15781541 TI - The effects of clonidine added to mepivacaine for paronychia surgery under axillary brachial plexus block. AB - We hypothesized that onset of sensory block is delayed in infected versus healthy tissues within the same nerve distribution after axillary brachial plexus block (ABPB) and that clonidine added to mepivacaine would enhance anesthesia and postoperative analgesia. Forty-one outpatients undergoing thumb/index paronychia surgery under ABPB were randomly assigned to receive in a double-blind fashion 400 mg mepivacaine plus either 100 microg clonidine (clonidine group, n = 21) or 2 mL saline (placebo group, n = 20). Onset of sensory block in the infected area was delayed compared with healthy areas of the same nerve distribution (24.7 +/- 5.5 min versus 21.3 +/- 7.2; P = 0.02 for median and 21.6 +/- 7.8 min; P = 0.04 for radial) within the placebo group. In the clonidine group, when compared to placebo i) onset of sensory block in both the median and radial nerve territories was accelerated (11.1 +/- 5.6 and 10.5 +/- 5.2 versus 21.3 +/- 7.2 and 21.6 +/- 7.8 min, respectively; P < 0.001), ii) onset of sensory block in the region of infection was accelerated (9.1 +/- 1.9 versus 24.7 +/- 5.5 min; P < 0.001), iii) duration of anesthesia (275 +/- 75 versus 163 +/- 57; P = 0.04) and time to first analgesic requirement (279 +/- 87 versus 197 +/- 84 min; P = 0.002) were prolonged with decreased visual analog scale scores at this time (30 +/- 18 versus 70 +/- 24; P < 0.001), and iv) verbal numeric rating scores were decreased at 24 h (1.7 +/- 2.2 versus 4.1 +/- 3.0; P = 0.002) and 48 h (0.1 +/- 0.5 versus 1.5 +/- 2.4; P = 0.01) postoperatively. Our findings suggest that in the setting of distal infected tissue surgery under ABPB infected tissues are resistant to anesthesia compared with healthy areas within the same nerve distribution and clonidine added to mepivacaine enhances both anesthesia and postoperative analgesia. PMID- 15781542 TI - A magnetic resonance imaging analysis of the infraclavicular region: can brachial plexus depth be estimated before needle insertion? AB - In this study we examined the anatomy of the infraclavicular region to assess the possibility of estimating brachial plexus depth before performing an infraclavicular block, by using readily identifiable landmarks such as the coracoid process (CP) and the clavicle (CL). Four parasagittal planes across the infraclavicular region were analyzed in 21 individual series of magnetic resonance imaging studies. Measurements included distance to the plexus from the skin of the anterior chest wall, position of the plexus relative to the CL, and clavicular width. The brachial plexus is located directly below the CL in the parasagittal plane 1 cm medial to the CP. If one inserts a needle in this same plane at a point in line with the inferomedial edge of the CP, then plexus depth can be estimated as follows. If the needle is raised, as a whole, straight up from the planned point of insertion to be level with the top of the CL, then the distance from the tip of the needle to a point midway across the width of the CL is equivalent to the distance from the insertion point to the plexus. Furthermore, not only is it uncommon to find the lung in this same parasagittal plane, but when it does appear, it is well behind the plexus. Estimating plexus depth, or "depth gauging," in the infraclavicular region is achievable and is a potentially useful strategy. Further study is required to confirm this finding in the clinical environment. PMID- 15781543 TI - Intravenous regional anesthesia using lidocaine and magnesium. AB - We conducted this study to evaluate the effects of magnesium, when added to lidocaine for IV regional anesthesia (IVRA), on tourniquet pain. Thirty patients undergoing elective hand surgery during IVRA were randomly assigned to two groups. IVRA was achieved with 10 mL of saline plus 3 mg/kg lidocaine 0.5% diluted with saline to a total of 40 mL in group C or with 10 mL of 15% magnesium sulfate (12.4 mmol) plus 3 mg/kg lidocaine 0.5% diluted with saline to a total of 40 mL in group M. Injection pain, sensory and motor block onset and recovery time, tourniquet pain, and anesthesia quality were noted. Patients were instructed to receive 75 mg of IM diclofenac when the visual analog scale (VAS) score was >4, and analgesic requirements were recorded. Sensory and motor block onset times were shorter and recovery times were prolonged in group M (P < 0.05). VAS scores of tourniquet pain were lower in group M at 15, 20, 30, 40, and 50 min (P < 0.001). Anesthesia quality, as determined by the anesthesiologist and surgeon, was better in group M (P < 0.05). Time to the first postoperative analgesic request in group C was 95 +/- 29 min and in group M was 155 +/- 38 min (P < 0.05). Postoperative VAS scores were higher for the first postoperative 6 h in group C (P < 0.05). Diclofenac consumption was significantly less in group M (50 +/- 35 mg) when compared with group C (130 + 55 mg) (P < 0.05). We conclude that magnesium as an adjunct to lidocaine improves the quality of anesthesia and analgesia in IVRA. PMID- 15781544 TI - The effects of stellate ganglion block on visual evoked potential and blood flow of the ophthalmic and internal carotid arteries in patients with ischemic optic neuropathy. AB - Ischemic optic neuropathy (ION) is a common disease that can cause a loss of visual acuity in the elderly. We treated ION patients with stellate ganglion block (SGB) and investigated its effects on picture visual evoked potential (P VEP) and blood flow in the ophthalmic artery (OA) and internal carotid artery (ICA). Twelve ischemic eyes in 12 patients diagnosed by the same ophthalmologist were investigated in this study. All patients were treated with daily SGB on the affected side with 2-3 mL of 2% lidocaine for a treatment period of 10-15 days. In ION eyes before SGB, compared with healthy eyes, the latency of P-VEP P(100) was delayed (123 +/- 14 ms versus 98 +/- 3 ms; P < 0.05), and the amplitude was reduced (4.24 +/- 1.76 microV versus 10.26 +/- 4.09 microV; P < 0.05). After SGB, the latency and amplitude returned to normal (103 +/- 6 ms versus 98 +/- 3 ms; 10.43 +/- 4.88 microV versus 10.26 +/- 4.09 microV; P > 0.05). Before treatment, the blood flow velocities of the OA and the ICA on the ischemic side were slow and the resistance indexes were high, but SGB reduced these changes. SGB did not affect the OA and the ICA on the healthy side. We conclude that SGB improves P VEP and OA and ICA blood flow in ION eyes. Further studies are needed to confirm that this is an effective method for the treatment of ION. PMID- 15781545 TI - Guillain-Barre syndrome in a patient with pancreatic cancer after an epidural general anesthetic. AB - Guillain-Barre syndrome is a rare occurrence in medicine and is probably rarer still as a postoperative complication. We report an uneventful operative course, during epidural-general anesthesia, in a patient undergoing pancreatectomy who presented with acute paralysis mimicking an acute cervical spinal cord syndrome or brachial plexus neuropathy. The signs and symptoms of right upper extremity paralysis occurred within 2 h postoperatively. Immediate work-up, which included magnetic resonance imaging, electromyography, and nerve conduction velocity studies, provided the diagnosis. PMID- 15781546 TI - The effects of cricoid pressure, remifentanil, and propofol on esophageal motility and the lower esophageal sphincter. AB - Cricoid pressure is the gold standard during the induction of anesthesia when there is a risk of aspiration of gastric contents. However, the effect of cricoid pressure during the different steps of complete anesthesia induction has not been studied. The purpose of this study was to investigate the effects of cricoid pressure, remifentanil, and propofol on lower esophageal sphincter (LES) and esophageal motility. We recorded LES pressure (LESP) and calculated barrier pressure ([BrP] = LESP - gastric pressure) in 10 healthy volunteers using a Dent sleeve device. There was a significant decrease in LESP and BrP when a cricoid pressure of 30 N was performed in the awake volunteers (P < 0.05). However, this effect was not seen during the infusion of remifentanil 0.2 microg . kg(-1) . min(-1). Remifentanil per se or together with a bolus dose of propofol 1 mg/kg IV did not induce any statistical change in LESP or BrP. Remifentanil abolished spontaneous esophageal motility and completely eliminated the experience of discomfort induced by cricoid pressure. In conclusion, cricoid pressure of 30 N induced a decrease of LESP and BrP in awake volunteers. These effects were not seen during the remifentanil infusion. This shows the importance of when to apply cricoid pressure during rapid-sequence induction. PMID- 15781547 TI - A comparison of sevoflurane-propofol versus sevoflurane or propofol for laryngeal mask airway insertion in adults. AB - In a prospective, randomized study, we investigated the incidence of successful insertion of laryngeal mask airway (LMA) at the first attempt and the incidence of side effects after LMA insertion using the combination of sevoflurane and propofol as compared with either sevoflurane or propofol alone for induction of anesthesia. Eighty-three unpremedicated ASA physical status I-II patients were anesthetized with a single vital capacity breath (VCB) of sevoflurane 8% supplemented with IV propofol 1.5 mg/kg, a single VCB of sevoflurane 8%, or IV propofol 3 mg/kg. The coinduction technique was associated with the most frequent incidence of successful LMA insertion at the first attempt (93.5%) than either sevoflurane alone (46%) or propofol alone (61.5%) (P < 0.001). Propofol-induced induction of anesthesia allowed the fastest insertion of LMA and was associated with the least frequent incidence of postoperative nausea and vomiting. However, this advantage of propofol was offset by a frequent incidence of pain on injection (69%) and the occurrence of movements during insertion of the LMA (50% in the propofol group versus 19% and 26% in the sevoflurane and sevoflurane propofol groups, respectively; P < 0.05), as well as a more frequent incidence of apnea (84% in the propofol group versus 7% and 16% in the sevoflurane and sevoflurane-propofol groups, respectively; P < 0.001). The report shows that induction of anesthesia with sevoflurane-propofol combined provides a frequent incidence of successful LMA insertion at the first attempt that is associated with an infrequent incidence of apnea. PMID- 15781548 TI - The chewing of betel quid and oral submucous fibrosis and anesthesia. AB - Oral submucous fibrosis (OSMF) is a premalignant lesion of the buccal mucosa caused by chewing betel quid. It results in progressive inability to open the mouth. OSMF causes difficulty in laryngoscopy and intubation of the trachea. Patients with OSMF require anesthesia for trismus correction, resection, and reconstructive (oncoplastic) surgery for coexisting oral malignancies or other unrelated surgeries. Our review of the anesthetics of 44 patients with oral malignancies suggested that 8 had OSMF. The preoperative airway assessment, including the Mallampati score and the clinical Tumor Node Metastasis stage, were useful in predicting the need for fiberoptic intubation. Patients with oral malignancies and OSMF had increased requirement for fiberoptic endotracheal intubations (62.5%) compared with those without OSMF (44.4%). Three different techniques of airway management (tracheal intubation after direct laryngoscopy, fiberoptic tracheal intubation, and tracheostomy) in four patients with OSMF are described. OSMF contributes to the development of the malignancy, delays the diagnoses, and complicates the anesthetic management. PMID- 15781549 TI - Regional anesthesia under general anesthesia and spinal cord injury. PMID- 15781550 TI - Asleep at the wheel? PMID- 15781551 TI - Exacerbation of carpal tunnel syndrome under treatment with valdecoxib. PMID- 15781553 TI - Cerebrospinal lavage seems to be safe and effective in the reversal of inadvertent spinal anesthetic injection. PMID- 15781555 TI - Intravascular catheter-associated infection. PMID- 15781557 TI - Reflex vagal withdrawal after sympathetic blockade. PMID- 15781558 TI - An unusual case of guidewire impaction during internal jugular venous cannulation. PMID- 15781559 TI - Bronchoconstriction induced by carotid sinus stimulation during radical neck dissection. PMID- 15781562 TI - Identification of the epidural space. PMID- 15781563 TI - Leukocytes with bright fluorescence in rats. PMID- 15781565 TI - Lightwand-assisted intubation of patients in the lateral decubitus position. PMID- 15781568 TI - Use of BIS monitoring was not associated with a reduced incidence of awareness. PMID- 15781569 TI - Subdural empyema combined with paraspinal abscess after epidural catheter insertion. PMID- 15781570 TI - Analysis of the floral transcriptome uncovers new regulators of organ determination and gene families related to flower organ differentiation in Gerbera hybrida (Asteraceae). AB - Development of composite inflorescences in the plant family Asteraceae has features that cannot be studied in the traditional model plants for flower development. In Gerbera hybrida, inflorescences are composed of morphologically different types of flowers tightly packed into a flower head (capitulum). Individual floral organs such as pappus bristles (sepals) are developmentally specialized, stamens are aborted in marginal flowers, petals and anthers are fused structures, and ovaries are located inferior to other floral organs. These specific features have made gerbera a rewarding target of comparative studies. Here we report the analysis of a gerbera EST database containing 16,994 cDNA sequences. Comparison of the sequences with all plant peptide sequences revealed 1656 unique sequences for gerbera not identified elsewhere within the plant kingdom. Based on the EST database, we constructed a cDNA microarray containing 9000 probes and have utilized it in identification of flower-specific genes and abundantly expressed marker genes for flower scape, pappus, stamen, and petal development. Our analysis revealed several regulatory genes with putative functions in flower-organ development. We were also able to associate a number of abundantly and specifically expressed genes with flower-organ differentiation. Gerbera is an outcrossing species, for which genetic approaches to gene discovery are not readily amenable. However, reverse genetics with the help of gene transfer has been very informative. We demonstrate here the usability of the gerbera microarray as a reliable new tool for identifying novel genes related to specific biological questions and for large-scale gene expression analysis. PMID- 15781571 TI - Disclosing hidden transcripts: mouse natural sense-antisense transcripts tend to be poly(A) negative and nuclear localized. AB - Genome-wide in silico analysis identified thousands of natural sense-antisense transcript (SAT) pairs in the mouse transcriptome. We investigated their expression using strand-specific oligo-microarray that distinguishes expression of sense and antisense RNA from 1947 SAT pairs. The majority of the predicted SATs are expressed at various steady-state levels in various tissues, and cluster analysis of the array data demonstrated that the ratio of sense and antisense expression for some of the SATs fluctuated markedly among these tissues, while the rest was unchanged. Surprisingly, further analyses indicated that vast amounts of multiple-sized transcripts are expressed from the SAT loci, which tended to be poly(A) negative, and nuclear localized. The tendency that the SATs are often not polyadenylated is conserved, even in the randomly chosen SAT genes in the plant Arabidopsis thaliana. Such common characteristics imply general roles of the SATs in regulation of gene expression. PMID- 15781573 TI - Chromosome triplication found across the tribe Brassiceae. AB - We have used an approximately 8.7-Mb BAC contig of Arabidopsis thaliana Chromosome 4 to trace homeologous chromosome regions in 21 species of the family Brassicaceae. Homeologs of this segment could be identified in all tested species. Painting of pachytene chromosomes of Calepina, Conringia, and Sisymbrium species (2n = 14, 16), traditionally placed in tribe Brassiceae, showed one homeologous copy of the Arabidopsis contig, while the remaining taxa of the tribe (2n = 14-30) revealed three, and three Brassica species (2n = 34, 36, and 38) and Erucastrum gallicum (2n = 30) had six copies corresponding to the 8.7-Mb segment. The multiple homeologous copies corresponded structurally to the Arabidopsis segment or were rearranged by inversions and translocations within the diploidized genomes. These chromosome rearrangements accompanied by chromosome fusions/fissions led to the present-day chromosome number variation within the Brassiceae. Phylogenetic relationships based on the chloroplast 5'-trnL (UAA) trnF(GAA) region and estimated divergence times based on sequence data of the chalcone synthase gene are congruent with comparative painting data and place Calepina, Conringia, and Sisymbrium outside the clade of Brassiceae species with triplicated genomes. Most likely, species containing three or six copy pairs descended from a common hexaploid ancestor with basic genomes similar to that of Arabidopsis. The presumed hexaploidization event occurred after the Arabidopsis Brassiceae split, between 7.9 and 14.6 Mya. PMID- 15781572 TI - The linkage disequilibrium maps of three human chromosomes across four populations reflect their demographic history and a common underlying recombination pattern. AB - The extent and patterns of linkage disequilibrium (LD) determine the feasibility of association studies to map genes that underlie complex traits. Here we present a comparison of the patterns of LD across four major human populations (African American, Caucasian, Chinese, and Japanese) with a high-resolution single nucleotide polymorphism (SNP) map covering almost the entire length of chromosomes 6, 21, and 22. We constructed metric LD maps formulated such that the units measure the extent of useful LD for association mapping. LD reaches almost twice as far in chromosome 6 as in chromosomes 21 or 22, in agreement with their differences in recombination rates. By all measures used, out-of-Africa populations showed over a third more LD than African-Americans, highlighting the role of the population's demography in shaping the patterns of LD. Despite those differences, the long-range contour of the LD maps is remarkably similar across the four populations, presumably reflecting common localization of recombination hot spots. Our results have practical implications for the rational design and selection of SNPs for disease association studies. PMID- 15781574 TI - Signaling for NKT cell development: the SAP-FynT connection. AB - New studies demonstrate a critical role for the adaptor protein SAP (SLAM associated protein) during NKT cell development. By connecting homotypic SLAM family receptor interactions with the FynT Src kinase, SAP may integrate a set of long-standing yet seemingly disparate observations characterizing NKT cell development. In fact, SAP-dependent signaling may underlie the development of multiple unconventional T cell lineages whose thymic selection relies on homotypic interactions between hematopoietic cells. PMID- 15781575 TI - T cell aging: naive but not young. AB - The immune system exhibits profound age-related changes, collectively termed immunosenescence. The most visible of these is the decline in protective immunity, which results from a complex interaction of primary immune defects and compensatory homeostatic mechanisms. The sum of these changes is a dysregulation of many processes that normally ensure optimal immune function. Recent advances suggest that old mice can produce fully functional new T cells, opening both intriguing inquiry avenues and raising critical questions to be pursued. PMID- 15781576 TI - Identifying genes that regulate bone remodeling as potential therapeutic targets. AB - Bone remodeling, a coupled process involving bone resorption and formation, is initiated by mechanical signals and is controlled by local and systemic factors that regulate osteoblast and osteoclast differentiation and function. An excess of resorption over formation leads to the bone loss and increased propensity to fracture that is characteristic of osteoporosis. A newly described inhibitor of osteoblast differentiation, Ciz, interferes with bone morphogenic protein signaling. As a consequence, Ciz-deficient mice develop increased bone mass. PMID- 15781577 TI - Newly generated CD4 T cells in aged animals do not exhibit age-related defects in response to antigen. AB - Using a T cell receptor transgenic (TCR Tg) mouse model, we have shown that TCR Tg CD4 cells from aged mice retain a naive phenotype, but exhibit reduced proliferation and IL-2 production in response to the antigen compared with cells from young mice. We hypothesize that age-related decreases in T cell function may be partly related to the age of the T cells. Because thymic output is decreased with age, peripheral T cells in older individuals are likely to be older than those in younger individuals. To investigate this possibility, we have manipulated the age of CD4 T cells in the periphery of young and aged mice. The production of new T cells was induced by depleting peripheral CD4 T cells or by creating bone marrow chimeras. In both young and aged individuals where we induced the production of new T cells, these newly generated cells exhibited robust responses to antigen ex vivo and in vivo, exhibiting good expansion, IL-2 production, and cognate helper function. Our results suggest that age-related defects in response to antigenic stimulation, in part, are caused by the age of the CD4 T cells. PMID- 15781578 TI - Granulocyte/macrophage colony-stimulating factor and accessory cells modulate radioprotection by purified hematopoietic cells. AB - Granulocyte/macrophage colony-stimulating factor (GM-CSF) promotes the survival, proliferation, and differentiation of myeloid lineage cells and regulates chemotaxis and adhesion. However, mice in which the genes encoding GM-CSF (Gmcsf) or the beta common subunit of the GM-CSF receptor (betac) are inactivated display normal steady-state hematopoiesis. Here, we show that host GM-CSF signaling strongly modulates the ability of donor hematopoietic cells to radioprotect lethally irradiated mice. Although bone marrow mononuclear cells efficiently rescue Gmcsf mutant recipients, fetal liver cells and Sca1(+) lin(-/dim) marrow cells are markedly impaired. This defect is partially attributable to accessory cells that are more prevalent in bone marrow. In contrast, Gmcsf-deficient hematopoietic stem cells demonstrate normal proliferative potentials. Short-term survival is also impaired in irradiated betac mutant recipients transplanted with fetal liver or bone marrow. These data demonstrate a nonredundant function of GM CSF in radioprotection by donor hematopoietic cells that may prove relevant in clinical transplantation. PMID- 15781579 TI - Platelet activation leads to activation and propagation of the complement system. AB - Inflammation and thrombosis are two responses that are linked through a number of mechanisms, one of them being the complement system. Various proteins of the complement system interact specifically with platelets, which, in turn, activates them and promotes thrombosis. In this paper, we show that the converse is also true: activated platelets can activate the complement system. As assessed by flow cytometry and immunoblotting, C3 deposition increased on the platelet surface upon cell activation with different agonists. Activation of the complement system proceeded to its final stages, which was marked by the increased generation of the anaphylotoxin C3a and the C5b-9 complex. We identified P-selectin as a C3b binding protein, and confirmed by surface plasmon resonance binding that these two proteins interact specifically with a dissociation constant of 1 microM. Using heterologous cells expressing P-selectin, we found that P-selectin alone is sufficient to activate the complement system, marked by increases in C3b deposition, C3a generation, and C5b-9 formation. In summary, we have found that platelets are capable of activating the complement system, and have identified P selectin as a receptor for C3b capable of initiating complement activation. These findings point out an additional mechanism by which inflammation may localize to sites of vascular injury and thrombosis. PMID- 15781580 TI - A mutation in the Icsbp1 gene causes susceptibility to infection and a chronic myeloid leukemia-like syndrome in BXH-2 mice. AB - BXH-2 mice develop a fatal myeloid leukemia by a two-step mutagenic process. First, a BXH-2-specific recessive mutation causes a myeloproliferative syndrome. Second, retroviral insertions alter oncogenes or tumor suppressors, resulting in clonal expansion of leukemic cells. We have identified a recessive locus on chromosome 8 (Myls) that is responsible for myeloproliferation in BXH-2. This Myls interval has been narrowed down to 2 Mb and found to contain several positional candidates, including the interferon consensus sequence-binding protein 1 gene (Icsbp, also known as interferon regulatory factor 8 [IRF8]). We show that BXH-2 mice carry a mutation (915 C to T) resulting in an arginine-to cysteine substitution at position 294 within the predicted IRF association domain of the protein. Although expression of Icsbp1 mRNA transcripts is normal in BXH-2 splenocytes, these cells are unable to produce interleukin 12 and interferon gamma in response to activating stimuli, confirming that R294C behaves as a loss of-function mutation. Myeloproliferation in BXH-2 mice is concomitant to increased susceptibility to Mycobacterium bovis (BCG) despite the presence of resistance alleles at the Nramp1 locus. These results suggest a two-step model for chronic myeloid leukemia in BXH-2, in which inactivation of Icsbp1 predisposes to myeloproliferation and immunodeficiency. This event is required for retroviral replication, and subsequent insertional mutagenesis that causes leukemia in BXH-2 mice. PMID- 15781581 TI - Transmission and accumulation of CTL escape variants drive negative associations between HIV polymorphisms and HLA. AB - Human immunodeficiency virus (HIV)-1 amino acid sequence polymorphisms associated with expression of specific human histocompatibility leukocyte antigen (HLA) class I alleles suggest sites of cytotoxic T lymphocyte (CTL)-mediated selection pressure and immune escape. The associations most frequently observed are between expression of an HLA class I molecule and variation from the consensus sequence. However, a substantial number of sites have been identified in which particular HLA class I allele expression is associated with preservation of the consensus sequence. The mechanism behind this is so far unexplained. The current studies, focusing on two examples of "negatively associated" or apparently preserved epitopes, suggest an explanation for this phenomenon: negative associations can arise as a result of positive selection of an escape mutation, which is stable on transmission and therefore accumulates in the population to the point at which it defines the consensus sequence. Such negative associations may only be in evidence transiently, because the statistical power to detect them diminishes as the mutations accumulate. If an escape variant reaches fixation in the population, the epitope will be lost as a potential target to the immune system. These data help to explain how HIV is evolving at a population level. Understanding the direction of HIV evolution has important implications for vaccine development. PMID- 15781582 TI - CD44 is a determinant of inflammatory bone loss. AB - Chronic inflammation is a major trigger of local and systemic bone loss. Disintegration of cell-matrix interaction is a prerequisite for the invasion of inflammatory tissue into bone. CD44 is a type I transmembrane glycoprotein that connects a variety of extracellular matrix proteins to the cell surface. Tumor necrosis factor (TNF) is a major inducer of chronic inflammation and its overexpression leads to chronic inflammatory arthritis. By generating CD44(-/-) human TNF-transgenic (hTNFtg) mice, we show that destruction of joints and progressive crippling is far more severe in hTNFtg mice lacking CD44, which also develop severe generalized osteopenia. Mutant mice exhibit an increased bone resorption due to enhanced number, size, and resorptive capacity of osteoclasts, whereas bone formation and osteoblast differentiation are not affected. Responsiveness of CD44-deficient osteoclasts toward TNF is enhanced and associated with increased activation of the p38 mitogen-activated protein kinase. These data identify CD44 as a critical inhibitor of TNF-driven joint destruction and inflammatory bone loss. PMID- 15781583 TI - Inhibition of platelet-derived growth factor signaling attenuates pulmonary fibrosis. AB - Pulmonary fibrosis is the consequence of a variety of diseases with no satisfying treatment option. Therapy-induced fibrosis also limits the efficacy of chemotherapy and radiotherapy in numerous cancers. Here, we studied the potential of platelet-derived growth factor (PDGF) receptor tyrosine kinase inhibitors (RTKIs) to attenuate radiation-induced pulmonary fibrosis. Thoraces of C57BL/6 mice were irradiated (20 Gy), and mice were treated with three distinct PDGF RTKIs (SU9518, SU11657, or Imatinib). Irradiation was found to induce severe lung fibrosis resulting in dramatically reduced mouse survival. Treatment with PDGF RTKIs markedly attenuated the development of pulmonary fibrosis in excellent correlation with clinical, histological, and computed tomography results. Importantly, RTKIs also prolonged the life span of irradiated mice. We found that radiation up-regulated expression of PDGF (A-D) isoforms leading to phosphorylation of PDGF receptor, which was strongly inhibited by RTKIs. Our findings suggest a pivotal role of PDGF signaling in the pathogenesis of pulmonary fibrosis and indicate that inhibition of fibrogenesis, rather than inflammation, is critical to antifibrotic treatment. This study points the way to a potential new approach for treating idiopathic or therapy-related forms of lung fibrosis. PMID- 15781584 TI - Asthmatic bronchial epithelial cells have a deficient innate immune response to infection with rhinovirus. AB - Rhinoviruses are the major trigger of acute asthma exacerbations and asthmatic subjects are more susceptible to these infections. To investigate the underlying mechanisms of this increased susceptibility, we examined virus replication and innate responses to rhinovirus (RV)-16 infection of primary bronchial epithelial cells from asthmatic and healthy control subjects. Viral RNA expression and late virus release into supernatant was increased 50- and 7-fold, respectively in asthmatic cells compared with healthy controls. Virus infection induced late cell lysis in asthmatic cells but not in normal cells. Examination of the early cellular response to infection revealed impairment of virus induced caspase 3/7 activity and of apoptotic responses in the asthmatic cultures. Inhibition of apoptosis in normal cultures resulted in enhanced viral yield, comparable to that seen in infected asthmatic cultures. Examination of early innate immune responses revealed profound impairment of virus-induced interferon-beta mRNA expression in asthmatic cultures and they produced >2.5 times less interferon-beta protein. In infected asthmatic cells, exogenous interferon-beta induced apoptosis and reduced virus replication, demonstrating a causal link between deficient interferon-beta, impaired apoptosis and increased virus replication. These data suggest a novel use for type I interferons in the treatment or prevention of virus-induced asthma exacerbations. PMID- 15781585 TI - A role for fungal {beta}-glucans and their receptor Dectin-1 in the induction of autoimmune arthritis in genetically susceptible mice. AB - A combination of genetic and environmental factors can cause autoimmune disease in animals. SKG mice, which are genetically prone to develop autoimmune arthritis, fail to develop the disease under a microbially clean condition, despite active thymic production of arthritogenic autoimmune T cells and their persistence in the periphery. However, in the clean environment, a single intraperitoneal injection of zymosan, a crude fungal beta-glucan, or purified beta-glucans such as curdlan and laminarin can trigger severe chronic arthritis in SKG mice, but only transient arthritis in normal mice. Blockade of Dectin-1, a major beta-glucan receptor, can prevent SKG arthritis triggered by beta-glucans, which strongly activate dendritic cells in vitro in a Dectin-1-dependent but Toll like receptor-independent manner. Furthermore, antibiotic treatment against fungi can prevent SKG arthritis in an arthritis-prone microbial environment. Multiple injections of polyinosinic-polycytidylic acid double-stranded RNA also elicit mild arthritis in SKG mice. Thus, specific microbes, including fungi and viruses, may evoke autoimmune arthritis such as rheumatoid arthritis by stimulating innate immunity in individuals who harbor potentially arthritogenic autoimmune T cells as a result of genetic anomalies or variations. PMID- 15781586 TI - The nucleocytoplasmic shuttling protein CIZ reduces adult bone mass by inhibiting bone morphogenetic protein-induced bone formation. AB - Osteoporosis is a major health problem; however, the mechanisms regulating adult bone mass are poorly understood. Cas-interacting zinc finger protein (CIZ) is a nucleocytoplasmic shuttling protein that localizes at cell adhesion plaques that form where osteoblasts attach to substrate. To investigate the potential role of CIZ in regulating adult bone mass, we examined the bones in CIZ-deficient mice. Bone volume was increased and the rates of bone formation were increased in CIZ deficient mice, whereas bone resorption was not altered. CIZ deficiency enhanced the levels of mRNA expression of genes encoding proteins related to osteoblastic phenotypes, such as alkaline phosphatase (ALP) as well as osterix mRNA expression in whole long bones. Bone marrow cells obtained from the femora of CIZ-deficient mice revealed higher ALP activity in culture and formed more mineralized nodules than wild-type cells. CIZ deficiency enhanced bone morphogenetic protein (BMP) induced osteoblastic differentiation in bone marrow cells in cultures, indicating that BMP is the target of CIZ action. CIZ deficiency increased newly formed bone mass after femoral bone marrow ablation in vivo. Finally, BMP-2-induced bone formation on adult mouse calvariae in vivo was enhanced by CIZ deficiency. These results establish that CIZ suppresses the levels of adult bone mass through inhibition of BMP-induced activation of osteoblasts. PMID- 15781589 TI - Differential gene expression profiling in whole blood during acute systemic inflammation in lipopolysaccharide-treated rats. AB - Microarrays have been used to evaluate the expression of thousands of genes in various tissues. However, few studies have investigated the change in gene expression profiles in one of the most easily accessible tissues, whole blood. We utilized an acute inflammation model to investigate the possibility of using a cDNA microarray to measure the gene expression profile in the cells of whole blood. Blood was collected from male Sprague-Dawley rats at 2 and 6 h after treatment with 5 mg/kg (ip) LPS. Hematology showed marked neutrophilia accompanied by lymphopenia at both time points. TNF-alpha and IL-6 levels were markedly elevated at 2 h, indicating acute inflammation, but by 6 h the levels had declined. Total RNA was isolated from whole blood and hybridized to the National Institute of Environmental Health Sciences Rat Chip v.3.0. LPS treatment caused 226 and 180 genes to be differentially expressed at 2 and 6 h, respectively. Many of the differentially expressed genes are involved in inflammation and the acute phase response, but differential expression was also noted in genes involved in the cytoskeleton, cell adhesion, oxidative respiration, and transcription. Real-time RT-PCR confirmed the differential regulation of a representative subset of genes. Principal component analysis of gene expression discriminated between the acute inflammatory response apparent at 2 h and the observed recovery underway at 6 h. These studies indicate that, in whole blood, changes in gene expression profiles can be detected that are reflective of inflammation, despite the adaptive shifts in leukocyte populations that accompany such inflammatory processes. PMID- 15781588 TI - The bovine PPARGC1A gene: molecular characterization and association of an SNP with variation of milk fat synthesis. AB - Several studies in a variety of breeds have reported at least two QTL for milk production traits, including milk fat synthesis on bovine chromosome 6 (BTA6), comprising a region that comparatively has been mapped to equivalent syntenic chromosome intervals in human, pig, and mouse harboring loci associated with type II diabetes and obesity-related traits. We identified the bovine peroxysome proliferator-activated receptor-gamma coactivator-1alpha gene (PPARGC1A) as a plausible positional and functional candidate gene for a previously described QTL for milk fat yield on BTA6 because of its chromosomal position and its key role in energy, fat, and glucose metabolism. To analyze the role of the bovine PPARGC1A gene in regulation of milk fat synthesis in dairy cattle, we determined its cDNA sequence, genomic organization, chromosomal localization, and expression pattern. The bovine PPARGC1A gene is organized in 13 exons comprising 6,261 bp and is expressed at different levels in a large number of tissues. Bovine PPARGC1A cDNA and protein sequences showed substantial similarity (92-95%) to its respective orthologs from human, rat, and mouse. Screening for polymorphisms in the coding sequence, exon/intron boundaries, 5'- and 3'-untranslated regions, and promoter region of the PPARGC1A gene in sires with a different genotype at the QTL for milk fat yield as well as in a multibreed panel revealed a total of 11 polymorphic loci. A significant association between an SNP in intron 9 of the PPARGC1A gene and milk fat yield was observed in a major dairy cattle population, indicating that the PPARGC1A gene could be involved in genetic variation underlying the QTL for milk fat synthesis on BTA6. PMID- 15781587 TI - In vivo depletion of lung CD11c+ dendritic cells during allergen challenge abrogates the characteristic features of asthma. AB - Although dendritic cells (DCs) play an important role in sensitization to inhaled allergens, their function in ongoing T helper (Th)2 cell-mediated eosinophilic airway inflammation underlying bronchial asthma is currently unknown. Here, we show in an ovalbumin (OVA)-driven murine asthma model that airway DCs acquire a mature phenotype and interact with CD4(+) T cells within sites of peribronchial and perivascular inflammation. To study whether DCs contributed to inflammation, we depleted DCs from the airways of CD11c-diphtheria toxin (DT) receptor transgenic mice during the OVA aerosol challenge. Airway administration of DT depleted CD11c(+) DCs and alveolar macrophages and abolished the characteristic features of asthma, including eosinophilic inflammation, goblet cell hyperplasia, and bronchial hyperreactivity. In the absence of CD11c(+) cells, endogenous or adoptively transferred CD4(+) Th2 cells did not produce interleukin (IL)-4, IL-5, and IL-13 in response to OVA aerosol. In CD11c-depleted mice, eosinophilic inflammation and Th2 cytokine secretion were restored by adoptive transfer of CD11c(+) DCs, but not alveolar macrophages. These findings identify lung DCs as key proinflammatory cells that are necessary and sufficient for Th2 cell stimulation during ongoing airway inflammation. PMID- 15781590 TI - Moving beyond the rhetoric of family-centered care. PMID- 15781591 TI - Mother's support options during cesarean childbirth. PMID- 15781592 TI - Abused pregnant women's interactions with health care providers during the childbearing year. AB - OBJECTIVE: To explore how intimate partner abuse during pregnancy influences women's decisions about seeking care and disclosing abuse and their preferences for health care professionals' responses. DESIGN: A qualitative, grounded theory approach using dimensional analysis. Data were collected via 21 interviews. SETTING: Participants were interviewed in their homes, at their prenatal clinic, at a hospital, and at the investigator's office. PARTICIPANTS: A convenience sample of 12 English-speaking women abused during pregnancy. Five participants were recruited from two prenatal clinics in the Pacific Northwest; 7 were recruited via snowball sampling. RESULTS: The grounded theory "living two lives: women's experiences of intimate partner abuse during pregnancy" emerged from the investigation. Abused pregnant women engaged in a process of guarding and revealing their public and private lives. This process affects how women seek and attend prenatal care, their perceptions of health care providers' interventions, and abuse disclosure. CONCLUSIONS: Intimate partner abuse profoundly affects women's pregnancies and pregnancy-related decisions. Embarrassment, shame, and fear are common emotions. Pregnant, abused women want health care providers to treat them respectfully and empathetically, to recognize the conflict between their public and private lives, to support their decisions, and to be available to help them. They do not expect or want health care providers to fix their situation. PMID- 15781593 TI - Postpartum symptoms after antepartum bed rest. AB - OBJECTIVE: To determine the type and frequency of postpartum symptoms during recovery from antepartum bed rest treatment across 6 weeks postpartum. DESIGN: Longitudinal repeated measures study. SETTING: Three perinatal tertiary care hospitals in two cities in the Midwest. PARTICIPANTS: One hundred six postpartum women who had a singleton high-risk pregnancy and were treated with antepartum bed rest. MAIN OUTCOME MEASURES: A Postpartum Symptom Checklist assessed physiologic and psychological symptoms. RESULTS: Women reported 12.5 symptoms at 2 days postpartum, but symptoms rapidly declined at 1 week. Ninety-three percent reported a mean of 6.6 symptoms at 6 weeks postpartum. Symptoms that continued to be reported at 6 weeks by at least 40% of women were fatigue, mood changes, tenseness, difficulty concentrating, back muscle soreness, dry skin, and headache. Women who had a cesarean delivery reported significantly more symptoms than those who had a vaginal delivery (p = .006). Length of maternal bed rest was significantly correlated with the number of symptoms at postpartum weeks 1, 2, 4, 5, and 6. CONCLUSION: Postpartum symptoms decreased across time but reveal an underlying morbidity that is not resolved by 6 weeks postpartum. PMID- 15781594 TI - The impact of hyperemesis gravidarum on maternal role assumption. AB - OBJECTIVES: To describe what it is like to have hyperemesis gravidarum and explore its impact on the assumption of the maternal role during the perinatal period. DESIGN: Qualitative descriptive design. PARTICIPANTS: Eight women who had been diagnosed with hyperemesis gravidarum were recruited through private prenatal clinics using a snowball technique. Participants were interviewed about their experiences with hyperemesis gravidarum in their homes or other suitable sites selected by them. MAIN OUTCOME MEASURES: Transcribed interviews were analyzed using grounded theory methodology. A core category and subcategories were determined. RESULTS: Hyperemesis gravidarum is a disorder that alters the usual response to pregnancy and birth. Participants in this study described it as so debilitating that they had little concern for anything else, including the fetus. After symptoms subsided, the participants reported that they were able to regain control over their lives and made an effort to make up for lost time in becoming attached to the baby. CONCLUSION: Women suffering from hyperemesis gravidarum may not benefit from the usual prenatal education efforts. Additional support following delivery may be needed as the woman attaches to her infant and learns to provide care. PMID- 15781595 TI - Meeting the challenge of new fatherhood during the early weeks. AB - OBJECTIVE: To explore new/subsequent Australian fathers' perspectives on the experiences, processes, and life changes in the early weeks of fatherhood. DESIGN: Interpretive study using in-depth interviews and grounded theory analysis techniques, based on a symbolic interactionist framework. SETTING: Participants were recruited from the postnatal wards of a major public hospital, early discharge program, and early childhood centers in southeast Queensland, Australia. PARTICIPANTS: Eighteen first-time/subsequent fathers interviewed 6 to 12 weeks after the birth. RESULTS: Although rewarding, fathers found new or expanding fatherhood to be a significant challenge and time of change. Major themes included making a commitment, taking responsibility, negotiating responsibilities, developing and maintaining relationships, maintaining family integrity, balancing activities, and perceiving the self as father. Work had a major impact on fathers' ability to participate with their family and newborn. To manage, fathers sought to balance the demands of work and home, deal with stressors, manage their time, develop routines, and reprioritize. Fathers developed a sense of themselves as fathers over time, building confidence and deriving satisfaction from their fathering role. CONCLUSIONS: A range of competing factors affected fathers' ability to participate in the home with their newborn in the early weeks after birth. PMID- 15781596 TI - Becoming an involved father of an infant. AB - DATA OBJECTIVE: To describe the experience of fatherhood in the early months after the birth of an infant through a metasynthesis of relevant qualitative studies. DATA SOURCES: Cinahl, PsychInfo, MEDLINE, and Social Work Abstracts electronic databases from 1990 through 2001 were searched using the terms qualitative, fathers, fatherhood, infants, father-infant relationship, and postpartum. STUDY SELECTION: Ten published articles, representing seven qualitative studies, focusing on the experiences of fathers of healthy infants were reviewed. Only studies published from 1990 through 2001 were selected to reflect more recent representations of fatherhood. DATA EXTRACTION AND SYNTHESIS: Using Noblit and Hare's metasynthesis approach, each study was carefully read, and key metaphors from each study were compared and translated by applying each of the metaphors to all the other studies. The study translations were synthesized into a whole, and the synthesis refined, leading to a description of the experience of being the father of an infant. CONCLUSIONS: The metasynthesis revealed that fathers of infants experienced four phases, represented by the following characteristics: (a) entering with expectations and intentions, (b) confronting reality, (c) creating one's role of involved father, and (d) reaping rewards. Contextual factors that influenced the father-infant relationship were identified. Implications for theory development, research, and clinical practice are discussed. PMID- 15781597 TI - Duration of breastfeeding for mothers of twins. AB - OBJECTIVE: To describe the incidence and duration of breastfeeding for mothers of twins and identify factors that affected this duration. DESIGN: Secondary analysis of data from a larger longitudinal predictive study of maternal attachment in mothers of twins. SETTING: Paper and pencil questionnaires once during pregnancy and twice in the first 6 months postpartum. PARTICIPANTS: 123 women recruited from a national support group for mothers of twins. MAIN OUTCOME MEASURES: Mothers' Information Tool, Edinburgh Postnatal Depression Scale, Rosenberg Self-Esteem Scale, and the Index of Breastfeeding Status. RESULTS: 110 (89.4%) of the sample initiated breastfeeding or initiated a milk supply by pumping. Percentage of breast milk feedings at time 2 predicted whether or not a woman was still breastfeeding at time 3 (odds ratio = 3.63, p < .001). CONCLUSION: A high percentage of breastfeeding initiation was found despite the increased care burden that has been described for mothers of twins. Mothers who continued to breastfeed at time 3 provided a high percentage of the twins' milk feedings as breast milk. The results suggest that mothers who are able to persist with the difficulties of establishing a milk supply for twins and feeding two infants are able to continue providing a high percentage of the infants' feedings as breast milk. PMID- 15781598 TI - Getting to know you: mothers' experiences of kangaroo care. AB - OBJECTIVE: To reveal mothers' experiences of providing kangaroo care for their preterm newborns while still in the hospital. DESIGN: Transcendental phenomenology was used to analyze the experiences of mothers providing kangaroo care for their preterm newborns. Tape recorded, semistructured interviews were conducted 1 to 4 weeks postpartum. Mothers were asked one grand tour question, "What was it like for you to provide kangaroo care for your preterm infant while in the hospital?" This study was the qualitative component of a randomized clinical trial. PARTICIPANTS: Ten women who provided kangaroo care for their preterm newborns, 32-36 completed weeks, weighing 1500-3000 grams, with APGAR scores 6 or greater at 1 minute, 7 or greater at 5 minutes. RESULTS: Four dominant themes emerged. The themes were reduced to one essential structure of knowing. The two essential elements of the structure of knowing were mothers kept from knowing their preterm newborn and mothers getting to know their preterm newborn. CONCLUSIONS: Kangaroo care facilitates bonding and enhances maternal infant acquaintance, even in the neonatal intensive care unit (NICU) environment. Mothers found that kangaroo care calmed them and their newborns. PMID- 15781599 TI - Implications of kangaroo care for growth and development in preterm infants. AB - OBJECTIVE: To review research on kangaroo care with implications for growth and development in preterm infants. DATA SOURCES: Nursing, medical, and child development research literature was searched through PubMed through 2003 using the search terms kangaroo Care, skin-to-skin, growth/development, and premature infants. STUDY SELECTION: Randomized controlled trials, pretest-posttest designs, and other comparative studies of kangaroo care were reviewed. Reports exploring parent perspectives were examined for attachment and parent-infant interaction findings. Theory and research regarding growth in preterm infants were explored. DATA EXTRACTION: Research on topics of kangaroo care, skin-to-skin contact, preterm infant growth, preterm infant weight gain, and failure to thrive was evaluated. DATA SYNTHESIS: Research on kangaroo care reports physiologic safety for preterm infants and increased attachment for parents. Attachment promotes nurturing behaviors that support growth and development. Weight gain as a benefit of kangaroo care remains in question. CONCLUSIONS: Kangaroo care is safe for preterm infants and may have important benefits for growth and development. Suggestions are made for future research on effects of KC on preterm infants. PMID- 15781600 TI - A psycho-educational program for improving women's attitudes and coping with menopause symptoms. AB - OBJECTIVE: To examine the impact of participation in a psycho-educational program on women's attitudes toward menopause, the perceived severity of their symptoms, and the association between the two. DESIGN: Quasi-experimental. Data were gathered at baseline and 3 months after termination of the program. SETTING: Two health maintenance organization clinics. PARTICIPANTS: Eighty-two healthy 40- to 60-year-old women who chose to participate in the study. Thirty-six women participated in the program, and 46 women comprised the control group. INTERVENTION: Program participants met for 10 weekly sessions to receive information from a professional team on subjects related to menopause and to share and discuss their experiences. MAIN OUTCOME MEASURES: Attitudes toward menopause and severity of menopause symptoms. RESULTS: The more negative the attitudes, the higher was the severity of symptoms. Participants reported significant improvements in attitudes and reductions in symptom severity compared to their own baseline scores and compared with the control group. CONCLUSIONS: Participation in a program that combines delivery of information and processing of experiences on the cognitive, emotional, and social levels may improve women's attitudes toward menopause and ease the perceived severity of their symptoms, thereby increasing quality of life. PMID- 15781602 TI - Women's Mental Health. PMID- 15781603 TI - Recognizing women's common mental health problems: the earthquake assessment model. AB - Women's reproductive events may coincide with the onset of a mood or anxiety disorder or the exacerbation of a previously existing one due to the interconnection of neurotransmitters, stress, and reproductive hormones. The women's health nurse plays a critical role in the identification of mood and anxiety disorders during a woman's life span. This article provides nurses with a mental health assessment model, describes the common mood and anxiety disorders, and discusses simple management and referral strategies. PMID- 15781604 TI - Reproductive health in women with eating disorders. AB - Medical complications are often the precipitants that lead women with eating disorders to seek health care. Complications can be wide ranging but frequently include symptoms associated with reproductive health. Unfortunately, because of the denial, embarrassment, shame, and secrecy associated with these psychiatric illnesses, the underlying cause of these complications can often go unacknowledged, delaying assessment and intervention. This article provides an overview of anorexia nervosa and bulimia nervosa, identifies the reproductive health issues that may occur in women with these disorders, and discusses the associated clinical implications for nursing practice. PMID- 15781605 TI - Identifying and treating postpartum depression. AB - Postpartum depression affects 10% to 20% of women in the United States and negatively influences maternal, infant, and family health. Assessment of risk factors and depression symptoms is needed to identify women at risk for postpartum depression for early referral and treatment. Individual and group psychotherapy have demonstrated efficacy as treatments, and some complementary/alternative therapies show promise. Treatment considerations include severity of depression, whether a mother is breastfeeding, and mother's preference. Nurses who work with childbearing women can advise depressed mothers regarding treatment options, make appropriate recommendations, provide timely and accessible referrals, and encourage engagement in treatment. PMID- 15781606 TI - Mood, energy, cognition, and physical complaints: a mind/body approach to symptom management during the climacteric. AB - Recent studies suggest that energy and cognitive and physical complaints such as fatigue, disrupted sleep, concentration problems, and pain are more problematic than clinical depression during the climacteric and may contribute to depressed mood. Central nervous system pathways that mediate mood, cognition, and energy are influenced by fluctuations of circulating estrogen during perimenopause. Symptoms are also influenced by other factors, including psychosocial and environmental stresses and supports. Health care for women from perimenopause to postmenopause should include an accurate assessment of energy and cognitive, physical, and emotional symptoms. Multidisciplinary approaches that combine prevention, symptom management, and health promotion are most effective for women. A mind/body program for perimenopause and menopause is presented as an example of a comprehensive treatment approach for assessment and management of perimenopause and menopause. This 10-week group program combined information, self-education, relaxation training, group support, lifestyle modification, and psychological coping skills. PMID- 15781607 TI - A psychiatric clinical nurse specialist as liaison to OB/GYN practice. AB - This article illustrates a model of liaison psychiatric nursing with an OB/GYN department to support the recognition and treatment of mental health problems in women. Education, prevention of illness, and early and effective treatment of disorders are emphasized. Key elements of an effective liaison role are discussed. PMID- 15781608 TI - Autophagy-dependent cell survival and cell death in an autosomal dominant familial neurohypophyseal diabetes insipidus in vitro model. AB - Mutations in the human gene encoding the antidiuretic hormone vasopressin (VP) cause autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI), a rare inherited disorder that presents as polydipsia and polyuria as a consequence of a loss of secretion of VP from posterior pituitary nerve terminals. Work from our laboratories has shown that adFNDI, like other neurodegenerative diseases such as Alzheimer's, Parkinson's and Huntington's, is associated with autophagy. We have recently shown that the activation of autophagy in mouse neuroblastoma Neuro2a cells after adenoviral vector-mediated delivery of an adFNDI mutant VP transgene (Cys67stop) is a cell survival mechanism; its inhibition induces apoptosis. We now show that expression of Cys67stop sensitizes Neuro2a cells to the lethal effects of dopamine. This mode of cell death exhibits features typically associated with classical apoptosis. Yet inhibition of autophagy reversed these effects and rescued cell viability. We propose that autophagy mediated cell death is a "two-hit" process: Following the cellular stress of the accumulation of a misfolded mutant protein, autophagy is prosurvival. However, a second insult triggers an autophagy-dependent apoptosis. PMID- 15781609 TI - Autophagy is a prosurvival mechanism in cells expressing an autosomal dominant familial neurohypophyseal diabetes insipidus mutant vasopressin transgene. AB - Autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI) is a progressive, inherited neurodegenerative disorder that presents as polydipsia and polyuria as a consequence of a loss of secretion of the antidiuretic hormone vasopressin (VP) from posterior pituitary nerve terminals. VP gene mutations cause adFNDI. Rats expressing an adFNDI VP transgene (Cys67stop) show a neuronal pathology characterized by autophagic structures in the cell body. adFNDI has thus been added to the list of protein aggregation diseases, along with Alzheimer's, Parkinson's and Huntington's, which are associated with autophagy, a bulk process that delivers regions of cytosol to lysosomes for degradation. However, the role of autophagy in these diseases is unclear. To address the relationships between mutant protein accumulation, autophagy, cell survival, and cell death, we have developed a novel and tractable in vitro system. We have constructed adenoviral vectors (Ads) that express structural genes encoding either the Cys67stop mutant protein (Ad-VCAT-Cys67stop) or an epitope-tagged wild type VP precursor (Ad-VCAT). After infection of mouse neuroblastoma Neuro2a cells, Ad-VCAT encoded material enters neurite processes and accumulates in terminals, while the Cys67stop protein is confined to enlarged vesicles in the cell body. Similar to the intracellular derangements seen in the Cys67stop rats, these structures are of ER origin, and colocalize with markers of autophagy. Neither Ad-VCAT-Cys67stop nor Ad-VCAT expression affected cell viability. However, inhibition of autophagy or lysosomal protein degradation, while having no effect on Ad-VCAT-expressing cells, significantly increased apoptotic cell death following Ad-VCAT-Cys67stop expression. These data suggest that activation of autophagy by the stress of the expression of an adFNDI mutant protein is a prosurvival mechanism. PMID- 15781610 TI - ABL oncogenes and phosphoinositide 3-kinase: mechanism of activation and downstream effectors. AB - The BCR-ABL oncogene is responsible for most cases of chronic myelogenous leukemia and some acute lymphoblastic leukemias. The fusion protein encoded by BCR-ABL possesses an aberrantly regulated tyrosine kinase activity. Imatinib mesylate (Gleevec, STI-571) is an inhibitor of ABL tyrosine kinase activity that has been remarkably effective in slowing disease progression in patients with chronic phase chronic myelogenous leukemia, but the emergence of imatinib resistance underscores the need for additional therapies. Targeting signaling pathways activated by BCR-ABL is a promising approach for drug development. The study of signaling components downstream of BCR-ABL and the related murine oncogene v-Abl has revealed a complex web of signals that promote cell division and survival. Of these, activation of phosphoinositide 3-kinase (PI3K) has emerged as one of the essential signaling mechanisms in ABL leukemogenesis. This review describes molecular mechanisms by which PI3K is activated and the downstream PI3K effectors that propagate the signal to promote myeloid and lymphoid transformation. Of particular recent interest is the mammalian target of rapamycin, a PI3K-regulated kinase that regulates protein synthesis and contributes to leukemogenesis. PMID- 15781611 TI - Lower induction of p53 and decreased apoptosis in NQO1-null mice lead to increased sensitivity to chemical-induced skin carcinogenesis. AB - NAD(P)H:quinone oxidoreductase 1 (NQO1) is a cytosolic protein that catalyzes metabolic detoxification of quinones and protects cells against redox cycling and oxidative stress. NQO1-null mice deficient in NQO1 protein showed increased sensitivity to 7,12-dimethylbenz(a)anthracene- and benzo(a)pyrene-induced skin carcinogenesis. In the present studies, we show that benzo(a)pyrene metabolite benzo(a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide and not benzo(a)pyrene quinones contributed to increased benzo(a) pyrene-induced skin tumors in NQO1-null mice. An analysis of untreated skin revealed an altered intracellular redox state due to accumulation of NADH and reduced levels of NAD/NADH in NQO1-null mice as compared with wild-type mice. Treatment with benzo(a)pyrene failed to significantly increase p53 and apoptosis in the skin of NQO1-null mice when compared with wild-type mice. These results led to the conclusion that altered intracellular redox state along with lack of induction of p53 and decreased apoptosis plays a significant role in increased sensitivity of NQO1-null mice to benzo(a)pyrene-induced skin cancer. PMID- 15781612 TI - The antifolate activity of tea catechins. AB - A naturally occurring gallated polyphenol isolated from green tea leaves, (-) epigallocatechin gallate (EGCG), has been shown to be an inhibitor of dihydrofolate reductase (DHFR) activity in vitro at concentrations found in the serum and tissues of green tea drinkers (0.1-1.0 micromol/L). These data provide the first evidence that the prophylactic effect of green tea drinking on certain forms of cancer, suggested by epidemiologic studies, is due to the inhibition of DHFR by EGCG and could also explain why tea extracts have been traditionally used in "alternative medicine" as anticarcinogenic/antibiotic agents or in the treatment of conditions such as psoriasis. EGCG exhibited kinetics characteristic of a slow, tight-binding inhibitor of 7,8-dihydrofolate reduction with bovine liver DHFR (K(I) = 0.109 micromol/L), but of a classic, reversible, competitive inhibitor with chicken liver DHFR (K(I) = 10.3 micromol/L). Structural modeling showed that EGCG can bind to human DHFR at the same site and in a similar orientation to that observed for some structurally characterized DHFR inhibitor complexes. The responses of lymphoma cells to EGCG and known antifolates were similar, that is, a dose-dependent inhibition of cell growth (IC50 = 20 micromol/L for EGCG), G0-G1 phase arrest of the cell cycle, and induction of apoptosis. Folate depletion increased the sensitivity of these cell lines to antifolates and EGCG. These effects were attenuated by growing the cells in a medium containing hypoxanthine-thymidine, consistent with DHFR being the site of action for EGCG. PMID- 15781613 TI - Cell type-specific tumor suppression by Ink4a and Arf in Kras-induced mouse gliomagenesis. AB - Homozygous deletion of the INK4a-ARF locus is one of the most frequent mutations found in human glioblastoma. We have previously shown that combined Ink4a-Arf loss can increase tumor incidence in both glial progenitor cells and astrocytes during mouse gliomagenesis. Here we have investigated the separate contribution of loss of each of the tumor suppressor genes in glial progenitor cells and astrocytes in Akt + Kras-induced gliomagenesis. We show that Arf is the major tumor suppressor gene in both cell types. Arf loss generated glioblastomas from both nestin-expressing glial progenitor cells and glial fibrillary acidic protein expressing astrocytes, with a significantly higher incidence in astrocytes. Ink4a loss, on the other hand, could only significantly contribute to gliomagenesis from glial progenitor cells and the induced tumors were of lower malignancy than those seen in Arf-deficient mice. Thus, Ink4a and Arf have independent and differential tumor suppressor functions in vivo in the glial cell compartment. PMID- 15781614 TI - Genetic and pharmacologic identification of Akt as a mediator of brain-derived neurotrophic factor/TrkB rescue of neuroblastoma cells from chemotherapy-induced cell death. AB - Patients whose neuroblastoma tumors express high levels of brain-derived neurotrophic factor (BDNF) and TrkB have an unfavorable prognosis. Our previous studies indicated that BDNF activation of the TrkB signal transduction pathway blocked the cytotoxic effects of chemotherapeutic drugs via the phosphatidylinositol 3-kinase pathway. Akt is an important downstream target of phosphatidylinositol 3-kinase and functions to regulate cell survival, proliferation, and protein synthesis. In this study, we examined whether Akt is required and sufficient to mediate BDNF/TrkB protection of neuroblastoma cells from chemotherapy. Transient transfection of a constitutively active Akt (Akt Myr) into TrkB-expressing SY5Y cells (TB8 cells) increases Akt activation and attenuates the cell death induced by chemotherapeutic reagents in the absence of BDNF. Furthermore, expression of a dominant-negative Akt (Akt-K179A) blocks the ability of BDNF to rescue TB8 cells from chemotherapy-induced cell death. Pharmacologic inhibition of Akt, with PIA6, a phosphatidylinositol ether lipid analogue (PIA), blocks BDNF-induced phosphorylation of Akt and the downstream target of Akt. PIA6 sensitizes neuroblastoma cells to chemotherapy and attenuates BDNF protection of neuroblastoma cells from chemotherapy-induced cell death. These results indicate that Akt is a key signaling component by which BDNF activation of the TrkB signal transduction pathway protects neuroblastoma cells from chemotherapy-induced cell death. This study raises the possibility that novel pharmacologic inhibitors of Akt may enhance the effectiveness of chemotherapeutic agents in the treatment of neuroblastoma tumors. PMID- 15781615 TI - Glycogen synthase kinase-3beta participates in nuclear factor kappaB-mediated gene transcription and cell survival in pancreatic cancer cells. AB - Recent studies using glycogen synthase kinase-3beta (GSK-3beta)-deficient mouse embryonic fibroblasts suggest that GSK-3beta positively regulates nuclear factor kappaB (NFkappaB)-mediated gene transcription. Because NFkappaB is suggested to participate in cell proliferation and survival pathways in pancreatic cancer, we investigated the role of GSK-3beta in regulating these cellular processes. Herein, we show that pancreatic cancer cells contain a pool of active GSK-3beta and that pharmacologic inhibition of GSK-3 kinase activity using small molecule inhibitors or genetic depletion of GSK-3beta by RNA interference leads to decreased cancer cell proliferation and survival. Mechanistically, we show that GSK-3beta influences NFkappaB-mediated gene transcription at a point distal to the Ikappa kinase complex, as only ectopic expression of the NFkappaB subunits p65/p50, but not an Ikappa kinase beta constitutively active mutant, could rescue the decreased cellular proliferation and survival associated with GSK-3beta inhibition. Taken together, our results simultaneously identify a previously unrecognized role for GSK-3beta in cancer cell survival and proliferation and suggest GSK-3beta as a potential therapeutic target in the treatment of pancreatic cancer. PMID- 15781616 TI - Androgen causes growth suppression and reversion of androgen-independent prostate cancer xenografts to an androgen-stimulated phenotype in athymic mice. AB - Most prostate cancer patients develop androgen-independent recurrent prostate tumors a few years after androgen ablation therapy. No therapy, however, has been shown to substantially extend survival in these patients. Previously, we reported that androgen suppresses the growth of androgen-independent LNCaP prostate tumor cells both in vitro and in vivo. In cell culture, androgen receptor (AR)-rich androgen-independent LNCaP 104-R1 cells adapt to growth suppression by androgen and then their growth is androgen stimulated. Because maintaining androgen dependency of prostate tumor cells should prolong the usefulness of androgen ablation therapy, we determined if androgen-independent prostate tumors would revert to an androgen-stimulated phenotype in vivo upon androgen treatment. Growth of the LNCaP 104-R1 tumors was suppressed by androgen, but tumors then adapted to suppression by androgen and growth became androgen stimulated. Tumor AR and prostate-specific antigen mRNA and protein were initially high in 104-R1 tumors but decreased during adaptation. Subsequent removal of androgen decreased the serum prostate-specific antigen level further and stopped the growth of the adapted tumors. Because androgen caused growth suppression and then reversion of androgen-independent tumors to an androgen-stimulated phenotype and because the growth of androgen-stimulated tumors could be restrained by androgen ablation, these results suggest a novel therapy for AR-positive androgen-independent prostate cancer. PMID- 15781617 TI - Activated Ras enhances insulin-like growth factor I induction of vascular endothelial growth factor in prostate epithelial cells. AB - Mutations in the three closely related RAS genes, HRAS, KRAS, and NRAS are among the most common mutations found in human cancer; reaching 50% in some types of cancer, such as colorectal carcinoma, and 10% in prostate cancers. The activated Ras proteins produced by these mutations can, among other cellular changes, increase vascular endothelial growth factor (VEGF) production. Moreover, tumors bearing RAS gene mutations are more vascular than tumors without RAS mutations. We find that, in prostate epithelial cells, the introduction of an activated HRAS causes cells to produce VEGF in response to insulin-like growth factor I (IGF-I). In comparison, cells lacking an activated Ras are unable to produce VEGF in response to IGF-I. This effect of Ras may occur through stabilization of a second messenger protein, insulin receptor substrate 1, that mediates PI 3-kinase dependent signaling. Because IGF-I is a paracrine/endocrine hormone that has been associated with increased risk for several types of cancer, these results suggest a novel interrelationship between oncogenic conversion of a cellular gene such as HRAS, and IGF-I produced locally for normal tissue homeostasis. PMID- 15781618 TI - Extensive chromosomal instability in Rad51d-deficient mouse cells. AB - Homologous recombination is a double-strand break repair pathway required for resistance to DNA damage and maintaining genomic integrity. In mitotically dividing vertebrate cells, the primary proteins involved in homologous recombination repair are RAD51 and the five RAD51 paralogs, RAD51B, RAD51C, RAD51D, XRCC2, and XRCC3. In the absence of Rad51d, human and mouse cells fail to proliferate, and mice defective for Rad51d die before birth, likely as a result of genomic instability and p53 activation. Here, we report that a p53 deletion is sufficient to extend the life span of Rad51d-deficient embryos by up to 6 days and rescue the cell lethal phenotype. The Rad51d-/- Trp53-/- mouse embryo-derived fibroblasts were sensitive to DNA-damaging agents, particularly interstrand cross links, and exhibited extensive chromosome instability including aneuploidy, chromosome fragments, deletions, and complex rearrangements. Additionally, loss of Rad51d resulted in increased centrosome fragmentation and reduced levels of radiation-induced RAD51-focus formation. Spontaneous frequencies of sister chromatid exchange were not affected by the absence of Rad51d, but sister chromatid exchange frequencies did fail to be induced upon challenge with the DNA cross-linking agent mitomycin C. These findings support a crucial role for mammalian RAD51D in normal development, recombination, and maintaining mammalian genome stability. PMID- 15781619 TI - C-Myc-independent restoration of multiple phenotypes by two C-Myc target genes with overlapping functions. AB - C-MYC, a transforming oncogene that is frequently overexpressed in many human cancers, regulates a variety of normal functions including cell cycle progression, apoptosis, and maintenance of cell size, morphology, and genomic integrity. Many target genes are modulated by c-Myc, and some can recapitulate a limited number of the above functions. Because most of these have been assessed in cells which also express endogenous c-Myc, however, it is not clear to what extent its proper regulation is also required. We show here that, in c-Myc nullizygous cells, two direct target genes, MT-MC1 and HMG-I, could each recapitulate multiple c-Myc phenotypes. Although these differ somewhat for the two genes, substantial overlap and cooperativity exist. The enforced expression of these two genes was also associated with the differential deregulation of some previously described c-Myc target genes, indicating the presence of a complex molecular circuitry. These observations argue that, despite the great diversity of gene regulation by c-Myc, many, although not all, of its functions can be phenocopied by a small subset of key downstream target genes. The approach described here should permit the identification of other target genes capable of further c-Myc-independent complementation. PMID- 15781620 TI - Lack of correlation between p53-dependent transcriptional activity and the ability to induce apoptosis among 179 mutant p53s. AB - Tumor suppressor p53-dependent apoptosis is thought to be one of the most important tumor-suppressive functions in human tumorigenesis. However, whether the major mechanism underlying the p53-dependent apoptosis is transactivation dependent or independent remains unclear. Using 179 mutant p53s with diverse transcriptional activities for distinct p53-binding sequences in yeast, we evaluated both their sequence-specific transcriptional activities on six p53 target genes and their ability to induce apoptosis in Saos-2 cells. These mutant p53s also represented diversity in their ability to both transactivate target genes and induce apoptosis. We identified 17 mutant p53s with superior ability to induce apoptosis than wild-type p53 that tend to cluster at residues 121 or 290 to 292. There was no significant correlation between the two functional properties on any single target gene examined. Furthermore, the 17 mutant p53s were not classified in a specific cluster by hierarchical cluster analysis on their diverse transcriptional activities, indicating that these mutant p53s were not similar in the transcriptional activity of downstream genes. These results suggested that transactivation-dependent apoptosis does not always play a major role in p53-dependent apoptosis, indirectly supporting the importance role of the transactivation-independent mechanism. PMID- 15781621 TI - Discovery of aberrant expression of R-RAS by cancer-linked DNA hypomethylation in gastric cancer using microarrays. AB - Although hypomethylation was the originally identified epigenetic change in cancer, it was overlooked for many years in preference to hypermethylation. Recently, gene activation by cancer-linked hypomethylation has been rediscovered. However, in gastric cancer, genome-wide screening of the activated genes has not been found. By using microarrays, we identified 1,383 gene candidates reactivated in at least one cell line of eight gastric cancer cell lines after treatment with 5-aza-2'deoxycytidine and trichostatin A. Of the 1,383 genes, 159 genes, including oncogenes ELK1, FRAT2, R-RAS, RHOB, and RHO6, were further selected as gene candidates that are silenced by DNA methylation in normal stomach mucosa but are activated by DNA demethylation in a subset of gastric cancers. Next, we showed that demethylation of specific CpG sites within the first intron of R-RAS causes activation in more than half of gastric cancers. Introduction of siRNA into R-RAS-expressing cells resulted in the disappearance of the adhered cells, suggesting that functional blocking of the R-RAS-signaling pathway has great potential for gastric cancer therapy. Our extensive gene list provides other candidates for this class of oncogene. PMID- 15781622 TI - Apoptosis factor EI24/PIG8 is a novel endoplasmic reticulum-localized Bcl-2 binding protein which is associated with suppression of breast cancer invasiveness. AB - p53 is a critical tumor suppressor which removes cells with DNA damage by regulating expression and activity of a select group of p53-induced genes (PIG) that subsequently induce apoptosis. PIG8 was also identified as a gene induced by etoposide and named etoposide-induced gene 24 (EI24). Later experiments established EI24/PIG8 as a proapoptotic factor and suggested that it may function as a tumor suppressor. Indeed, EI24/PIG8 is relatively highly mutated in aggressive breast cancers and is located in a region which expresses frequent loss of heterozygosity. However, despite these important observations, the activity and role of EI24/PIG8 remain largely unknown. We used (immmuno)fluorescence microscopy and subcellular fractionation techniques to show that EI24/PIG8 is localized in the endoplasmic reticulum (ER). Pull-down experiments showed that it specifically binds with Bcl-2, a death regulator known to reside in mitochondria, ER, and the nuclear envelope. EI24/PIG8-Bcl-2 binding was corroborated by coimmunoprecipitation and other in vitro and in vivo protein protein binding assays. Further analysis showed that EI24/PIG8 uses its N terminal region to bind the BH3 domain in Bcl-2. Finally, we used immunohistochemical techniques to analyze expression of EI24/PIG8 in breast cancer tissue progression arrays and showed that loss of EI24/PIG8 is associated with tumor invasiveness but not with the development of the primary tumor. These results suggest that EI24/PIG8 is a novel, ER-localized Bcl-2-binding protein which may contribute to apoptosis by modulating the activity and/or function of Bcl-2 in this organelle. EI24/PIG8 may serve to prevent tumor spreading, consistent with its suspected role as a tumor suppressor. PMID- 15781623 TI - Functional Loss of the gamma-catenin gene through epigenetic and genetic pathways in human prostate cancer. AB - Gamma-catenin is a cell adhesion molecule and a candidate mediator of Wnt signal transduction. We hypothesized that impaired regulation of gamma-catenin through genetic and epigenetic pathways is associated with the pathogenesis of prostate cancer. To test this hypothesis, cytosine-phosphate-guanine methylation, loss of heterozygosity (LOH), and mutation status of the gamma-catenin gene were analyzed in cultured prostate cancer cell lines, 180 localized prostate cancers, 69 benign prostatic hyperplasias, and 11 hormone refractory prostate cancers (HRPC). In prostate cancer cell lines (DuPro, LNCaP, ND-1, and PC3), gamma-catenin mRNA transcripts were increased after 5-aza-2'-deoxycytidine treatment. In localized prostate cancer, gamma-catenin expression was lower but prevalence of gamma catenin methylation was higher compared with benign prostatic hyperplasia. However, gamma-catenin methylation did not correlate with Gleason sum, pT category, or capsular penetration. Among localized prostate cancers with positive gamma-catenin methylation, the presence of LOH at chromosome 17q21 was closely related to down-regulation of gamma-catenin mRNA expression. The gamma-catenin mutations were not found in localized prostate cancers, whereas six mutations were found in five HRPCs within or close to the GSK-3beta consensus motif phosphorylation site, among which four HRPCs showed strong nuclear gamma-catenin accumulation. In these four HRPCs, Bcl-2 expression was increased, whereas the target of the Wnt signal, c-myc, was only expressed in one HRPC. Therefore, although epigenetic gamma-catenin methylation is an early event in the development of prostate cancer, simultaneous events of epigenetic cytosine phosphate-guanine methylation and genetic LOH may be responsible for functional loss of gamma-catenin. The gamma-catenin mutation related to Bcl-2 overexpression has a significant effect on the pathogenesis of HRPC. This is the first report to characterize the epigenetic and genetic regulation of gamma-catenin in human prostate cancer. PMID- 15781624 TI - Loss of the inactive X chromosome and replication of the active X in BRCA1 defective and wild-type breast cancer cells. AB - In females, X chromosome inactivation (XCI) begins with the expression of the XIST gene from the X chromosome destined to be inactivated (Xi) and the coating of XIST RNA in cis. It has recently been reported that this process is supported by the product of the BRCA1 tumor suppressor gene and that BRCA1-/- cancers show Xi chromatin structure defects, thus suggesting a role of XCI perturbation in BRCA1-mediated tumorigenesis. Using a combined genetic and epigenetic approach, we verified the occurrence of XCI in BRCA1-/- and BRCA1wt breast cancer cell lines. It was ascertained that the Xi was lost in all cancer cell lines, irrespective of the BRCA1 status and that more than one active X (Xa) was present. In addition, no epigenetic silencing of genes normally subjected to XCI was observed. We also evaluated XIST expression and found that XIST may be occasionally transcribed also from Xa. Moreover, in one of the BRCA1wt cell line the restoring of XIST expression using a histone deacetylase inhibitor, did not lead to XCI. To verify these findings in primary tumors, chromosome X behavior was investigated in a few BRCA1-associated and BRCA1-not associated primary noncultured breast carcinomas and the results mirrored those obtained in cancer cell lines. Our findings indicate that the lack of XCI may be a frequent phenomenon in breast tumorigenesis, which occurs independently of BRCA1 status and XIST expression and is due to the loss of Xi and replication of Xa and not to the reactivation of the native Xi. PMID- 15781625 TI - Molecular staging of cervical lymph nodes in squamous cell carcinoma of the head and neck. AB - Clinical staging of cervical lymph nodes from patients with squamous cell carcinoma of the head and neck (SCCHN) has only 50% accuracy compared with definitive pathologic assessment. Consequently, both clinically positive and clinically negative patients frequently undergo neck dissections that may not be necessary. To address this potential overtreatment, sentinel lymph node (SLN) biopsy is currently being evaluated to provide better staging of the neck. However, to fully realize the potential improvement in patient care afforded by the SLN procedure, a rapid and accurate SLN analysis is necessary. We used quantitative reverse transcription-PCR (QRT-PCR) to screen 40 potential markers for their ability to detect SCCHN metastases to cervical lymph nodes. Seven markers were identified with good characteristics for identifying metastatic disease, and these were validated using a set of 26 primary tumors, 19 histologically positive lymph nodes, and 21 benign nodes from patients without cancer. Four markers discriminated between positive and benign nodes with accuracy >97% but only one marker, pemphigus vulgaris antigen (PVA), discriminated with 100% accuracy in both the observed data and a statistical bootstrap analysis. A rapid QRT-PCR assay for PVA was then developed and incorporated into a prototype instrument capable of performing fully automated RNA isolation and QRT-PCR. The automated analysis with PVA provided perfect discrimination between histologically positive and benign lymph nodes and correctly identified two lymph nodes with micrometastatic tumor deposits. These assays were completed (from tissue to result) in approximately 30 minutes, thus demonstrating the feasibility of intraoperative staging of SCCHN SLNs by QRT-PCR. PMID- 15781626 TI - Cytoplasmic HuR expression is a prognostic factor in invasive ductal breast carcinoma. AB - HuR is a ubiquitously expressed mRNA-binding protein. Intracellular localization of HuR is predominantly nuclear, but it shuttles between the nucleus and the cytoplasm. In the cytoplasm it can stabilize certain transcripts. Because nucleocytoplasmic translocation of HuR is necessary for its activity, it was hypothesized that cytoplasmic HuR expression in cancer cells could be a prognostic marker. To test the significance of HuR in carcinogenesis of the breast, we have investigated HuR expression in a mouse mammary gland tumor model and from 133 invasive ductal breast carcinoma specimens. HuR expression was elevated in the cyclooxygenase-2 transgene-induced mouse mammary tumors, and its expression was predominantly cytoplasmic in the tumor cells. In the human carcinoma samples, high cytoplasmic immunoreactivity for HuR was found in 29% (38 of 133) of the cases. Cytoplasmic HuR expression associated with high grade (P = 0.0050) and tumor size over 2 cm (P = 0.0082). Five-year distant disease-free survival rate was 42% [95% confidence interval (95% CI), 26-58] in cytoplasm-high category and 84% (95% CI, 76-91) in cytoplasm-negative or -low category (P < 0.0001), and high cytoplasmic expression of HuR was an independent prognostic factor in a Cox multivariate model (relative risk 2.07; 95% CI, 1.05-4.07). Moreover, high cytoplasmic HuR immunopositivity was significantly associated with poor outcome in the subgroup of node-negative breast cancer in a univariate analysis (P < 0.0007). Our results show that high cytoplasmic HuR expression is associated with a poor histologic differentiation, large tumor size, and poor survival in ductal breast carcinoma. Thus, HuR is the first mRNA stability protein of which expression associates with poor outcome in breast cancer. PMID- 15781627 TI - Human epididymis protein 4 (HE4) is a secreted glycoprotein that is overexpressed by serous and endometrioid ovarian carcinomas. AB - Among the genes most commonly identified in gene expression profiles of epithelial ovarian carcinomas (EOC) is the gene for human epididymis protein 4 (HE4). To ascertain its clinical utility, we did a comprehensive assessment of HE4 protein expression in benign and malignant ovarian and nonovarian tissues by immunohistochemistry. In comparison with normal surface epithelium, which does not express HE4, we found that cortical inclusion cysts lined by metaplastic Mullerian epithelium abundantly express the protein. Its expression in tumors was restricted to certain histologic subtype: 93% of serous and 100% of endometrioid EOCs expressed HE4, whereas only 50% and 0% of clear cell carcinomas and mucinous tumors, respectively, were positive. Tissue microarrays revealed that the majority of nonovarian carcinomas do not express HE4, consistent with our observation that HE4 protein expression is highly restricted in normal tissue to the reproductive tracts and respiratory epithelium. HE4 is predicted to encode a secreted protein. Using reverse transcription-PCR, we identified ovarian cancer cell lines that endogenously overexpress HE4. Cultured medium from these cells revealed a secreted form of HE4 that is N-glycosylated. This observation is consistent with the recent report that HE4 circulates in the bloodstream of patients with EOC. Therefore, HE4 is a secreted glycoprotein that is overexpressed by serous and endometrioid EOCs. Its expression in cortical inclusion cysts suggests that formation of Mullerian epithelium is a prerequisite step in the development of some types of EOCs. PMID- 15781628 TI - Gene expression profiling identifies molecular subtypes of inflammatory breast cancer. AB - Breast cancer is a heterogeneous disease. Comprehensive gene expression profiles obtained using DNA microarrays have revealed previously indistinguishable subtypes of noninflammatory breast cancer (NIBC) related to different features of mammary epithelial biology and significantly associated with survival. Inflammatory breast cancer (IBC) is a rare, particular, and aggressive form of disease. Here we have investigated whether the five molecular subtypes described for NIBC (luminal A and B, basal, ERBB2 overexpressing, and normal breast-like) were also present in IBC. We monitored the RNA expression of approximately 8,000 genes in 83 breast tissue samples including 37 IBC, 44 NIBC, and 2 normal breast samples. Hierarchical clustering identified the five subtypes of breast cancer in both NIBC and IBC samples. These subtypes were highly similar to those defined in previous studies and associated with similar histoclinical features. The robustness of this classification was confirmed by the use of both alternative gene set and analysis method, and the results were corroborated at the protein level. Furthermore, we show that the differences in gene expression between NIBC and IBC and between IBC with and without pathologic complete response that we have recently reported persist in each subtype. Our results show that the expression signatures defining molecular subtypes of NIBC are also present in IBC. Obtained using different patient series and different microarray platforms, they reinforce confidence in the expression-based molecular taxonomy but also give evidence for its universality in breast cancer, independently of a specific clinical form. PMID- 15781629 TI - Immortalization of primary human prostate epithelial cells by c-Myc. AB - A significant percentage of prostate tumors have amplifications of the c-Myc gene, but the precise role of c-Myc in prostate cancer is not fully understood. Immortalization of human epithelial cells involves both inactivation of the Rb/p16INK4a pathway and telomere maintenance, and it has been recapitulated in culture by expression of the catalytic subunit of telomerase, hTERT, in combination with viral oncoproteins. Here, we show the immortalization of human prostate epithelial cells (HPrEC) by a single genetic event, the expression of the c-Myc oncogene. Myc stabilizes telomere length in HPrEC through up-regulation of hTERT expression and overrides the accumulation of cell cycle inhibitory proteins, such as p16INK4a. Overall, HPrECs expressing c-Myc retain many characteristics of normal cells, such as the induction of a senescence-like growth arrest in response to oncogenic Ras, an intact p53 response, and an absence of gross karyotypic abnormalities. However, HPrECs expressing c-Myc lack a Rb/p16INK4a checkpoint and can be transformed without the need for additional genetic lesions in that pathway. These results give a partial explanation for the physiologic role of c-Myc overexpression in prostate cancer. PMID- 15781630 TI - p73beta-Mediated apoptosis requires p57kip2 induction and IEX-1 inhibition. AB - Similarly to p53, p73alpha and p73beta induce growth arrest and/or apoptosis in response to DNA damage or when exogenously expressed. However, how they trigger apoptosis remains unresolved. After stable transduction of either p73alpha or p73beta, a greater apoptotic response was observed for p73beta in both primary and tumor cells. Consistently, blocking ectopic and endogenous p73beta expression by specific shRNA significantly decreased apoptotic levels after DNA damage. We found that p73beta targets the apoptotic program at multiple levels: (i) facilitating caspase activation through p53-dependent signals and (ii) inducing p57KIP2, while down-regulating c-IPA1 and IEX1 through a p53-independent mechanism. p73beta-mediated apoptosis was considerably reduced after inhibition of p57(KIP2) by small interfering RNA, IEX-1 overexpression, and in mouse embryo fibroblasts derived from p57-/- mice. Data from this study offer evidence for the apoptotic activity exclusive of p73beta. In the clinical context, these results might have potential therapeutic implications, because p73beta could induce alternative apoptotic responses in tumors harboring p53 mutations. PMID- 15781631 TI - Expression of the carcinoembryonic antigen gene is inhibited by SOX9 in human colon carcinoma cells. AB - The human carcinoembryonic antigen (CEA) is overexpressed in many types of human cancers and is commonly used as a clinical marker. In colon cancer, this overexpression protects cells against apoptosis and contributes to carcinogenesis. Therefore, CEA-expressing cells as well as CEA expression itself constitute potential therapeutic targets. In this report, we show that the transcription factor SOX9 down-regulates CEA gene expression and, as a probable consequence, induces apoptosis in the human colon carcinoma cell line HT29Cl.16E. PMID- 15781632 TI - CDK4 and MDM2 gene alterations mainly occur in highly proliferative and aggressive mantle cell lymphomas with wild-type INK4a/ARF locus. AB - Amplification of 12q13 locus occurs in some mantle cell lymphomas (MCL), potentially involving CDK4 and MDM2 genes. To determine the role of these genes in MCL, we have examined their gene status and expression and their relationship to INK4a/ARF and p53 gene aberrations in 69 tumors. Increased CDK4 gene copy number was detected in 4 of 19 (21%) highly proliferative blastoid variants and was associated with mRNA and protein overexpression. Three additional cases showed mRNA overexpression with no structural alterations of the gene. MDM2 gene overexpression was detected in three blastoid tumors (16%) with no relationship to gene copy gains. INK4a/ARF and p53 aberrations were observed in 13 and 12 tumors, respectively. Four of the seven lymphomas with CDK4 aberrations had concurrent inactivation of p53 gene, whereas only one case had a concomitant homozygous deletion of INK4a/ARF. No other gene alterations were found in the three cases with MDM2 overexpression. Patients with INK4a/ARF deletions or simultaneous aberrations of p53 and CDK4 had a significantly shorter median survival (17 months) than patients with isolated alterations of p53, MDM2, or CDK4 (32 months) and patients with no alterations in any of these genes (77 months). The prognostic impact of the concomitant oncogenic alterations of the p14ARF/p53 and p16INK4a/CDK4 pathways was independent of the proliferation of the tumors. These findings indicate that CDK4 and MDM2 gene alterations mainly occur in MCL with a wild-type INK4a/ARF locus and may contribute to the higher proliferation and more aggressive behavior of the tumors. PMID- 15781633 TI - Cancer/testis antigen expression in human mesenchymal stem cells: down-regulation of SSX impairs cell migration and matrix metalloproteinase 2 expression. AB - Several families of genes by and large located on the X chromosome encode proteins of unspecified function. Commonly known as cancer/testis (CT) antigens, they are considered, under normal conditions, only to be expressed in cells of the germ line and placenta. CT genes are also often expressed in cancer cells, hence their classification. Here we report that their expression in normal cells is wider spread and can be observed in cells with the potential for self-renewal and pleuripotency, namely, stem cells. Several CT genes and their products, CT antigens, including SSX, NY-ESO-1, and N-RAGE, were expressed in undifferentiated mesenchymal stem cells (MSCs) and down-regulated after osteocyte and adipocyte differentiation. To elucidate the possible overlapping function played by these genes in cancer and stem cells, a comparative analysis of the localization of their proteins was made. In addition, localization relative to other MSC markers was examined. This revealed that SSX localizes in the cytoplasm and overlap occurs in regions where matrix metalloproteinase 2 (MMP2) and vimentin accumulate. Nevertheless, it was found that no protein interactions between these molecules occur. Further investigation revealed that the migration of a melanoma cell line (DFW), which expresses SSX, MMP2, and vimentin, decreases when SSX is down-regulated. This decrease in cell migration was paralleled by a reduction in MMP2 levels. Analogous to this, SSX expression is down-regulated in MSCs after differentiation; concomitantly a reduction in MMP2 levels occurs. In addition, E cadherin expression increases, mimicking a mesenchymal epithelial transition. These results afford SSX a functional role in normal stem cell migration and suggest a potentially similar function in cancer cell metastases. PMID- 15781634 TI - The E7 protein of cutaneous human papillomavirus type 8 causes invasion of human keratinocytes into the dermis in organotypic cultures of skin. AB - Human papillomaviruses (HPV) have been implicated in the development of nonmelanoma skin cancer (NMSC). The molecular mechanisms by which these viruses contribute towards NMSC are poorly understood. We have used an in vitro skin equivalent model generated by transducing primary adult human epidermal keratinocytes with retroviruses expressing HPV genes to investigate the mechanisms of viral transformation. In this model, keratinocytes expressing HPV genes are seeded onto a mesenchyme composed of deepidermalized human dermis that had been repopulated with primary dermal fibroblasts. Expression of the HPV8 E7 gene caused both an enhancement of terminal differentiation and hyperproliferation, but most strikingly, the acquisition of the ability to migrate and invade through the underlying dermis. The basement membrane integrity was disrupted in a time-dependent manner in areas of invading keratinocytes, as evidenced by immunostaining of its protein components collagen types VII, IV, and laminin 5. This was accompanied by the overexpression of extracellular matrix metalloproteinases MMP-1, MMP-8, and MT-1-MMP. These results suggest that the cutaneous HPV type 8 that is frequently found in NMSC of epidermodysplasia verruciformis patients may actively promote an invasive keratinocyte phenotype. These findings also highlight the importance of epithelial-extracellular matrix mesenchymal interactions that are required to support cell invasion. PMID- 15781635 TI - Expression and nuclear location of the transcriptional repressor Kaiso is regulated by the tumor microenvironment. AB - Kaiso is a BTB/POZ zinc finger protein originally described as an interaction partner of p120ctn. In cultured cell lines, Kaiso is found almost exclusively in the nucleus, where it generally acts as a transcriptional repressor. Here, we describe the first in situ immunolocalization studies of Kaiso expression in normal and cancerous tissues. Surprisingly, we found striking differences between its behavior in monolayers of different cell lines, three-dimensional cell culture systems, and in vivo. Although nuclear localization was sometimes observed in tissues, Kaiso was more often found in the cytoplasm, and in some cell types it was absent. In general, Kaiso and p120ctn did not colocalize in the nucleus. To examine this phenomenon more carefully, tumor cells exhibiting strong nuclear Kaiso staining in vitro were injected into nude mice and grown as xenografts. The latter showed a progressive translocation of Kaiso towards the cytoplasm over time, and even complete loss of expression, especially in the center of the tumor nodules. When xenografted tumors were returned to cell culture, Kaiso was re-expressed and was once again found in the nucleus. Translocation of Kaiso to the cytoplasm and down-regulation of its levels were also observed under particular experimental conditions in vitro, such as formation of spheroids and acini. These data strongly imply an unexpected influence of the microenvironment on Kaiso expression and localization. As transcriptional repression is a nuclear event, this phenomenon is likely a crucial factor in the regulation of Kaiso function. PMID- 15781636 TI - Cyclooxygenase-2 functions as a downstream mediator of lysophosphatidic acid to promote aggressive behavior in ovarian carcinoma cells. AB - Elevated levels of the bioactive lipid lysophosphatidic acid (LPA) are detectable in the majority of patients with both early- and late-stage ovarian cancer, suggesting that LPA promotes early events in ovarian carcinoma dissemination. LPA contributes to the development, progression, and metastasis of ovarian cancer in part by inducing the expression of genes that contribute to proliferation, survival, or invasion, including cyclooxgenase-2 (COX-2) and matrix metalloproteinase-2 (MMP-2). We have previously shown that LPA promotes proMMP-2 activation and MMP-2-dependent migration and invasion in ovarian cancer cells. The purpose of the current study was to determine whether the effect of LPA on acquisition of the metastatic phenotype in ovarian cancer cells is mediated via a COX-2-dependent mechanism. Immunohistochemical analysis of 173 ovarian tumors showed strong COX-2 immunoreactivity in 63% of tumor specimens, including 50% of borderline tumors. LPA increased COX-2 protein expression in a time- and concentration-dependent manner in two of three immortalized borderline ovarian epithelial cells as well as in four of six ovarian cancer cell lines. This was accomplished by both activation of the Edg/LPA receptor and LPA-mediated transactivation of the epidermal growth factor receptor, which increased COX-2 expression via the Ras/mitogen-activated protein kinase pathway. COX-2 also played a role in LPA-induced invasion and migration, as treatment with the COX-2 specific inhibitor NS-398 reduced LPA-induced proMMP-2 protein expression and activation and blocked MMP-dependent motility and invasive activity. These data show that COX-2 functions as a downstream mediator of LPA to potentiate aggressive cellular behavior. PMID- 15781637 TI - HYAL1 hyaluronidase: a molecular determinant of bladder tumor growth and invasion. AB - Hyaluronic acid and HYAL1-type hyaluronidase show high accuracy in detecting bladder cancer and evaluating its grade, respectively. Hyaluronic acid promotes tumor progression; however, the functions of hyaluronidase in cancer are largely unknown. In this study, we stably transfected HT1376 bladder cancer cells with HYAL1-sense (HYAL1-S), HYAL1-antisense (HYAL1-AS), or vector cDNA constructs. Whereas HYAL1-S transfectants produced 3-fold more HYAL1 than vector transfectants, HYAL1-AS transfectants showed approximately 90% reduction in HYAL1 production. HYAL1-AS transfectants grew four times slower than vector and HYAL1-S transfectants and were blocked in the G2-M phase of the cell cycle. The expression of cdc25c and cyclin B1 and cdc2/p34-associated H1 histone kinase activity also decreased in HYAL1-AS transfectants. HYAL1-S transfectants were 30% to 44% more invasive, and HYAL1-AS transfectants were approximately 50% less invasive than the vector transfectants in vitro. In xenografts, there was a 4- to 5-fold delay in the generation of palpable HYAL1-AS tumors, and the weight of HYAL1-AS tumors was 9- to 17-fold less than vector and HYAL1-S tumors, respectively (P < 0.001). Whereas HYAL1-S and vector tumors infiltrated skeletal muscle and blood vessels, HYAL1-AS tumors resembled benign neoplasia. HYAL1-S and vector tumors expressed significantly higher amounts of HYAL1 (in tumor cells) and hyaluronic acid (in tumor-associated stroma) than HYAL1-AS tumors. Microvessel density in HYAL1-S tumors was 3.8- and 9.5-fold higher than that in vector and HYAL1-AS tumors, respectively. These results show that HYAL1 expression in bladder cancer cells regulates tumor growth and progression and therefore serves as a marker for high-grade bladder cancer. PMID- 15781638 TI - Peroxisome proliferator-activated receptor gamma inhibition prevents adhesion to the extracellular matrix and induces anoikis in hepatocellular carcinoma cells. AB - Activation of the nuclear transcription factor peroxisome proliferator-activated receptor gamma (PPARgamma) inhibits growth and survival of hepatocellular carcinoma (HCC) cell lines. To further investigate the function of PPARgamma in HCC, PPARgamma expression patterns in primary tumors were examined, and the responses of two HCC cell lines to PPARgamma activation and inhibition were compared. PPARgamma expression was increased in HCC and benign-appearing peritumoral hepatocytes compared with remote benign hepatocytes. Both compound PPARgamma inhibitors and PPARgamma small interfering RNAs prevented HCC cell lines from adhering to the extracellular matrix. Loss of adhesion was followed by caspase-dependent apoptosis (anoikis). PPARgamma inhibitors had no effect on initial beta1 integrin-mediated adhesion, or on total focal adhesion kinase levels but did reduce focal adhesion kinase phosphorylation. The PPARgamma inhibitor T0070907 was significantly more efficient at causing cancer cell death than the activators troglitazone and rosiglitazone. T0070907 caused cell death by reducing adhesion and inducing anoikis, whereas the activators had no direct effect on adhesion and caused cell death at much higher concentrations. In conclusion, PPARgamma overexpression is present in HCC. Inhibition of PPARgamma function causes HCC cell death by preventing adhesion and inducing anoikis mediated apoptosis. PPARgamma inhibitors represent a potential novel treatment approach to HCC. PMID- 15781639 TI - Tbx2 is overexpressed and plays an important role in maintaining proliferation and suppression of senescence in melanomas. AB - The INK4a and ARF genes found at the CDKN2A locus are key effectors of cellular senescence that is believed to act as a powerful anticancer mechanism. Accordingly, mutations in these genes are present in a wide variety of spontaneous human cancers and CDKN2A germ line mutations are found in familial melanoma. The TBX2 gene encoding a key developmental transcription factor is amplified in pancreatic cancer cell lines and preferentially amplified and overexpressed in BRCA1 and BRCA2 mutated breast tumors. Overexpression of Tbx2 and the related factor Tbx3, which is also overexpressed in breast cancer and melanomas, can suppress senescence in defined experimental systems through repression of ARF expression. However, it is not known how Tbx2 mediates its repressive effect nor whether endogenous Tbx2 or Tbx3 perform a similar antisenescence function in transformed cells. This is a particularly important question because the loss of CDKN2A in many human cancers would, in principle, bypass the requirement for Tbx2/3-mediated repression of ARF in suppressing senescence. We show here that Tbx2 is overexpressed in melanoma cell lines and that Tbx2 targets histone deacetylase 1 to the p21Cip1 (CDKN1A) initiator. Strikingly, expression of an inducible dominant-negative Tbx2 (dnTbx2) leads to displacement of histone deacetylase 1, up-regulation of p21(Cip1) expression, and the induction of replicative senescence in CDKN2A-null B16 melanoma cells. In human melanoma cells, expression of dnTbx2 leads to severely reduced growth and induction of senescence-associated heterochromatin foci. The results suggest that the activity of endogenous Tbx2 is critically required to maintain proliferation and suppress senescence in melanomas. PMID- 15781640 TI - Dehydroepiandrosterone can inhibit the proliferation of myeloma cells and the interleukin-6 production of bone marrow mononuclear cells from patients with myeloma. AB - The serum levels of an adrenal sex hormone, dehydroepiandrosterone sulfate (DHEA S), are significantly more decreased in human myelomas compared with the reduction brought by physiologic decline with age. In order to clarify the effect of DHEA on myeloma cells, we investigated whether DHEA and DHEA-S could inhibit interleukin-6 (IL-6) production of bone marrow mononuclear cells and the proliferation of myeloma cells from patients with myeloma. DHEA-S and DHEA suppressed IL-6 production from a bone marrow stromal cell line, KM-102, as well as in bone marrow mononuclear cells from patients with myeloma. Furthermore, DHEA inhibited in vitro growth of the U-266 cell line and primary myeloma cells from the patients, as well as the in vivo growth of U-266 cells implanted i.p. in severe combined immunodeficiency-hIL6 transgenic mice. DHEA up-regulated the expression of peroxisome proliferator-activated receptor (PPAR), PPAR beta, but not PPARgamma or PPARalpha, and the expression of IkappaBalpha gene in myeloma cells and bone marrow stromal cells, which could explain the suppressive effect of DHEA on IL-6 production through the down-regulation of NF-kappaB activity. Therefore, these data revealed that DHEA-S, as well as DHEA, had a direct effect on myeloma and bone marrow stromal cells to inhibit their proliferation and IL-6 production, respectively. PMID- 15781641 TI - Targeted replacement of hypoxia-inducible factor-1alpha by a hypoxia-inducible factor-2alpha knock-in allele promotes tumor growth. AB - Hypoxia-inducible factors (HIF) are essential transcriptional regulators that mediate adaptation to hypoxic stress in rapidly growing tissues such as tumors. HIF activity is regulated by hypoxic stabilization of the related HIF-1alpha and HIF-2alpha subunits, which are frequently overexpressed in cancer cells. To assess the relative tumor-promoting functions of HIF-1alpha and HIF-2alpha directly, we replaced HIF-1alpha expression with HIF-2alpha by creating a novel "knock-in" allele at the Hif-1alpha locus through homologous recombination in primary murine embryonic stem cells. Compared with controls, s.c. teratomas derived from knock-in embryonic stem cells were larger and more proliferative, had increased microvessel density, and exhibited increased expression of vascular endothelial growth factor, transforming growth factor-alpha, and cyclin D1. These and other data indicate that HIF-2alpha promotes tumor growth more effectively than HIF-1alpha in multiple contexts. PMID- 15781642 TI - Emodin down-regulates androgen receptor and inhibits prostate cancer cell growth. AB - Hormone-refractory relapse is an inevitable and lethal event for advanced prostate cancer patients after hormone deprivation. A growing body of evidence indicates that hormone deprivation may promote this aggressive prostate cancer phenotype. Notably, androgen receptor (AR) not only mediates the effect of androgen on the tumor initiation but also plays the major role in the relapse transition. This provides a strong rationale for searching new effective agents targeting the down-regulation of AR to treat or prevent advanced prostate cancer progression. Here, we show that emodin, a natural compound, can directly target AR to suppress prostate cancer cell growth in vitro and prolong the survival of C3(1)/SV40 transgenic mice in vivo. Emodin treatment resulted in repressing androgen-dependent transactivation of AR by inhibiting AR nuclear translocation. Emodin decreased the association of AR and heat shock protein 90 and increased the association of AR and MDM2, which in turn induces AR degradation through proteasome-mediated pathway in a ligand-independent manner. Our work indicates a new mechanism for the emodin-mediated anticancer effect and justifies further investigation of emodin as a therapeutic and preventive agent for prostate cancer. PMID- 15781643 TI - Effect of conditional knockout of the type II TGF-beta receptor gene in mammary epithelia on mammary gland development and polyomavirus middle T antigen induced tumor formation and metastasis. AB - Transforming growth factor-beta (TGF-beta) isoforms are growth factors that function physiologically to regulate development, cellular proliferation, and immune responses. The role of TGF-beta signaling in mammary tumorigenesis is complex, as TGF-beta has been reported to function as both a tumor suppressor and tumor promoter. To elucidate the role of TGF-beta signaling in mammary gland development, tumorigenesis, and metastasis, the gene encoding type II TGF-beta receptor, Tgfbr2, was conditionally deleted in the mammary epithelium (Tgfbr2MGKO). Loss of Tgfbr2 in the mammary epithelium results in lobular alveolar hyperplasia in the developing mammary gland and increased apoptosis. Tgfbr2MGKO mice were mated to the mouse mammary tumor virus-polyomavirus middle T antigen (PyVmT) transgenic mouse model of metastatic breast cancer. Loss of Tgfbr2 in the context of PyVmT expression results in a shortened median tumor latency and an increased formation of pulmonary metastases. Thus, our studies support a tumor-suppressive role for epithelial TGF-beta signaling in mammary gland tumorigenesis and show that pulmonary metastases can occur and are even enhanced in the absence of TGF-beta signaling in the carcinoma cells. PMID- 15781644 TI - Human osteocalcin and bone sialoprotein mediating osteomimicry of prostate cancer cells: role of cAMP-dependent protein kinase A signaling pathway. AB - Osteocalcin and bone sialoprotein are the most abundant noncollagenous bone matrix proteins expressed by osteoblasts. Surprisingly, osteocalcin and bone sialoprotein are also expressed by malignant but not normal prostate epithelial cells. The purpose of this study is to investigate how osteocalcin and bone sialoprotein expression is regulated in prostate cancer cells. Our investigation revealed that (a) human osteocalcin and bone sialoprotein promoter activities in an androgen-independent prostate cancer cell line of LNCaP lineage, C4-2B, were markedly enhanced 7- to 12-fold in a concentration-dependent manner by conditioned medium collected from prostate cancer and bone stromal cells. (b) Deletion analysis of human osteocalcin and bone sialoprotein promoter regions identified cyclic AMP (cAMP)-responsive elements (CRE) as the critical determinants for conditioned medium-mediated osteocalcin and bone sialoprotein gene expression in prostate cancer cells. Consistent with these results, the protein kinase A (PKA) pathway activators forskolin and dibutyryl cAMP and the PKA pathway inhibitor H-89, respectively, increased or repressed human osteocalcin and bone sialoprotein promoter activities. (c) Electrophoretic mobility shift assay showed that conditioned medium-mediated stimulation of human osteocalcin and bone sialoprotein promoter activities occurs through increased interaction between CRE and CRE-binding protein. (d) Conditioned medium was found to induce human osteocalcin and bone sialoprotein promoter activities via increased CRE/CRE-binding protein interaction in a cell background-dependent manner, with marked stimulation in selected prostate cancer but not bone stromal cells. Collectively, these results suggest that osteocalcin and bone sialoprotein expression is coordinated and regulated through cAMP-dependent PKA signaling, which may define the molecular basis of the osteomimicry exhibited by prostate cancer cells. PMID- 15781645 TI - IGFBP-3 is a direct target of transcriptional regulation by DeltaNp63alpha in squamous epithelium. AB - DeltaNp63alpha is a nuclear transcription factor that maintains epithelial progenitor cell populations, is overexpressed in several epithelial cancers, and can negatively regulate apoptosis. However, the mechanisms by which DeltaNp63alpha promotes cell survival are unclear. DeltaNp63alpha has been reported to act as a transcriptional repressor, but specific target genes directly repressed by DeltaNp63alpha remain unidentified. Here, we present evidence that DeltaNp63alpha functions to negatively regulate the proapoptotic protein IGFBP-3. Disruption of p63 expression in squamous epithelial cells increases IGFBP-3 expression, whereas ectopic expression of DeltaNp63alpha down regulates IGFBP-3. DeltaNp63alpha binds to sites in the IGFBP-3 gene in vivo and can modulate transcription through these sites. Furthermore, DeltaNp63alpha and IGFBP-3 expression patterns are inversely correlated in normal squamous epithelium and squamous cell carcinomas. These data suggest that IGFBP-3 is a target of transcriptional repression by DeltaNp63alpha and that this repression represents a mechanism by which tumors that overexpress p63 may be protected from apoptosis. PMID- 15781646 TI - Reconstitution of human telomerase reverse transcriptase expression rescues colorectal carcinoma cells from in vitro senescence: evidence against immortality as a constitutive trait of tumor cells. AB - Although in vitro establishment of new colorectal carcinoma (CRC) cell lines is an infrequent event, we have observed that primary cultures of CRC can be repeatedly and reproducibly initiated following in vitro plating of tumor-derived epithelial cells. These cultures, however, usually display a short life span as they undergo a limited number of cell passages before entering a state of irreversible growth arrest. In this study, we show that short-lived CRC primary cultures lack constitutive telomerase activity and undergo a senescence process characterized by progressive telomere shortening. Moreover, transduction of these cells with a retroviral vector encoding human telomerase reverse transcriptase (hTERT) is sufficient to reconstitute telomerase activity and allow immortalization. Detailed molecular characterization of hTERT-immortalized CRC cell lines confirms their individual tumor origin by showing expression of colonic epithelial differentiation markers, such as cytokeratin-20 (CK20), full match with class I and class II human leukocyte antigen genotyping of autologous B-lymphoblastoid cells, and presence of somatic mutations in key cancer genes (KRAS2, APC) identical to those of the corresponding autologous original tumor tissues. Moreover, functional characterization of hTERT-immortalized CRC cell lines shows that they have a transformed phenotype, being able to form colonies in soft agar and tumors in severe combined immunodeficient mice. Most interestingly, immunohistochemical analysis of original tumor tissues indicates that short-lived CRC primary cultures, although hTERT-negative in vitro, derive from hTERT-positive tumors. Taken together, our data show that, in a least subset of CRC, biochemical pathways involved in maintenance of telomere length, such as telomerase, are not activated in a constitutive way in all tumor cells. PMID- 15781647 TI - The propeptide mediates formation of stromal stores of PROMIC-1: role in determining prostate cancer outcome. AB - The extracellular matrix (ECM) is a reservoir of cellular binding proteins and growth factors that are critical for normal cell behavior, and aberrations in the ECM invariably accompany malignancies such as prostate cancer. Carcinomas commonly overexpress macrophage inhibitory cytokine 1 (MIC-1), a proapoptotic and antitumorigenic transforming growth factor-beta superfamily cytokine. Here we show that MIC-1 is often secreted in an unprocessed propeptide containing form. It is variably processed intracellularly, with unprocessed forms being secreted from several tumor lines, including prostate carcinoma lines, PC-3 and LNCaP. Once secreted, only unprocessed proMIC-1 binds ECM, demonstrating for the first time the occurrence of extracellular stores of MIC-1. The propeptide mediates this association via its COOH-terminal 89 amino acids. Xenograft models bearing tumors secreting various engineered forms of MIC-1 show that the propeptide regulates the balance between ECM stores and circulating serum levels of mature MIC-1 in vivo. The absence of propeptide results in approximately 20-fold increase in serum MIC-1 levels. The significance of stromal MIC-1 stores was evaluated in prostate cancer tissue cores, which show major variation in stromal levels of MIC-1. Stromal MIC-1 levels are linked to prostate cancer outcome following radical prostatectomy, with decreasing stromal levels providing an important independent predictor of disease relapse. In low-grade localized prostate cancer (Gleason sum score < or = 6), the level of MIC-1 stromal stores was the best predictor of future relapse when compared with all other clinicopathologic variables. The secretion and ECM association of unprocessed proMIC-1 is likely to play a central role in modulating local bioavailability of MIC-1 which can affect patient outcome in prostate cancer and other epithelial tumors. PMID- 15781648 TI - Hair follicle-derived blood vessels vascularize tumors in skin and are inhibited by Doxorubicin. AB - We have recently shown that the neural-stem cell marker nestin is expressed in hair follicle stem cells and the blood vessel network interconnecting hair follicles in the skin of transgenic mice with nestin regulatory element-driven green fluorescent protein (ND-GFP). The hair follicles were shown to give rise to the nestin-expressing blood vessels in the skin. In the present study, we visualized tumor angiogenesis by dual-color fluorescence imaging in ND-GFP transgenic mice after transplantation of the murine melanoma cell line B16F10 expressing red fluorescent protein. ND-GFP was highly expressed in proliferating endothelial cells and nascent blood vessels in the growing tumor. Results of immunohistochemical staining showed that the blood vessel-specific antigen CD31 was expressed in ND-GFP-expressing nascent blood vessels. ND-GFP expression was diminished in the vessels with increased blood flow. Progressive angiogenesis during tumor growth was readily visualized during tumor growth by GFP expression. Doxorubicin inhibited the nascent tumor angiogenesis as well as tumor growth in the ND-GFP mice transplanted with B16F10-RFP. This model is useful for direct visualization of tumor angiogenesis and evaluation of angiogenic inhibitors. PMID- 15781649 TI - Predominant Bcl-XL knockdown disables antiapoptotic mechanisms: tumor necrosis factor-related apoptosis-inducing ligand-based triple chemotherapy overcomes chemoresistance in pancreatic cancer cells in vitro. AB - Pancreatic cancer is lethal because of its invasiveness, rapid progression, and profound resistance to chemotherapy and radiation therapy. To identify the molecular mechanisms underlying this, we have examined the expression and potency of three major death receptors: tumor necrosis factor receptor (TNF-R), TNF related apoptosis-inducing ligand receptor (TRAIL-R), and Fas in mediating cytotoxicity in four invasive pancreatic cancer cell lines. We have analyzed the expression of major antiapoptotic factors, cell cycle regulators and death receptor decoys (DcR) in comparison with normal pancreas tissues and five other human malignant tumor cell lines. We have found that different pancreatic cancer cell lines coexpress high-level TRAIL-R, Fas, and TNF-R1 but are strongly resistant to apoptosis triggered by the death receptors. DcR2 and DcR3 overexpression may partly contribute to the resistance of pancreatic cancer cells to TRAIL-R- and Fas-mediated cytotoxicity. Bcl-XL and Bcl-2 are predominantly overexpressed in pancreatic cancer cell lines, respectively. Bcl-XL is also predominantly overexpressed in prostate, colorectal, and intestinal cancer cells. The knockdown of the predominant Bcl-XL overexpression significantly reduces the viability of pancreatic cancer cells to TNFalpha- and TRAIL-mediated apoptosis by sublethal-dose single and combined antitumor drugs, including geldanamycin, PS 341, Trichostatin A, and doxorubicine. Geldanamyin and PS-341 synergistically block NFkappaB activation, suppress Akt/PKB pathway, and down-regulate Bcl-XL, Bcl-2, cIAP-1, and cyclin D1 expression. This combined regimen dramatically enhances TRAIL cytotoxic effects and breaks through chemoresistance. Bcl-XL plays a vital role in pancreatic cancer chemoresistance. Geldanamycin, PS-341, and TRAIL triple combination may be a novel therapeutic strategy for pancreatic cancer. PMID- 15781650 TI - Expression of Notch-1 and its ligands, Delta-like-1 and Jagged-1, is critical for glioma cell survival and proliferation. AB - The Notch family of proteins plays an integral role in determining cell fates, such as proliferation, differentiation, and apoptosis. We show that Notch-1 and its ligands, Delta-like-1 and Jagged-1, are overexpressed in many glioma cell lines and primary human gliomas. Immunohistochemistry of a primary human glioma tissue array shows the presence in the nucleus of the Notch-1 intracellular domain, indicating Notch-1 activation in situ. Down-regulation of Notch-1, Delta like-1, or Jagged-1 by RNA interference induces apoptosis and inhibits proliferation in multiple glioma cell lines. In addition, pretreatment of glioma cells with Notch-1 or Delta-like-1 small interfering RNA significantly prolongs survival in a murine orthotopic brain tumor model. These results show, for the first time, the dependence of cancer cells on a single Notch ligand; they also suggest a potential Notch juxtacrine/autocrine loop in gliomas. Notch-1 and its ligands may present novel therapeutic targets in the treatment of glioma. PMID- 15781651 TI - Modulation of survival signaling pathways and persistence of the genotoxic stress as a basis for the synergistic interaction between the atypical retinoid ST1926 and the epidermal growth factor receptor inhibitor ZD1839. AB - Strategies targeting apoptotic pathways may have relevance to improve the efficacy of antitumor therapy. Because synthetic atypical retinoids are potent inducers of apoptosis, there is an increasing interest in exploiting their potential in novel therapeutic approaches. In the present study, we have investigated the cellular effects of the combination of a novel atypical retinoid, ST1926, and the epidermal growth factor receptor inhibitor ZD1839. The results indicated a synergistic interaction between the two drugs associated with a dramatic enhancement of apoptotic response, up-regulation of the cell death receptor DR5, and caspase 8 activation. Other molecular events induced by the cotreatment included (a) a stabilization of the ST1926-induced genotoxic stress detected by formation of phosphorylated gamma-H2AX foci and (b) a complete inhibition of extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation associated with activation of the proapoptotic protein BAD (i.e., inhibition of phosphorylation on Ser112). In addition, ZD1839 itself inhibited survival pathways by causing a partial dephosphorylation of Akt and a marked down regulation of survivin. The role of ERK-mediated survival pathways in the cellular response to the drug combination was further supported by the counteracting effect of stimulation of survival pathways by an alternative receptor tyrosine kinase and by the use of a specific inhibitor of the ERK pathway. In conclusion, the results support that the survival pathways activated by epidermal growth factor receptor are determinants of the cell susceptibility to ST1926-induced apoptosis and lowering survival signals may increase the cellular sensitivity to the atypical retinoid. The favorable pharmacologic profiles of both ST1926 and ZD1839 suggest that the combination of these well tolerated agents may have therapeutic potential. PMID- 15781652 TI - Selective apoptotic killing of malignant hemopoietic cells by antibody-targeted delivery of an amphipathic peptide. AB - The alpha-helical amphipathic peptide D-(KLAKLAK)2 is toxic to eukaryotic cells if internalized by a suitable targeting mechanism. We have targeted this peptide to malignant hemopoietic cells via conjugation to monoclonal antibodies, which recognize lineage-specific cell surface molecules. An anti-CD19/peptide conjugate efficiently killed 3/3 B lymphoid lines. However, an anti-CD33/peptide conjugate was cytotoxic to only one of three CD33-positive myeloid leukemia lines. The IC50 towards susceptible lines were in the low nanomolar range. Conjugates were highly selective and did not kill cells that did not express the appropriate cell surface cognate of the antibody moiety. Anti-CD19/peptide conjugates efficiently killed cells from patients with chronic lymphocytic leukemia but anti CD33/peptide reagents were less effective against fresh acute myeloid leukemia cells. We therefore suggest that amphipathic peptides may be of value as targeted therapeutic agents for the treatment of a subset of hematologic malignancies. PMID- 15781653 TI - Increased expression of apoptosis inhibitor protein XIAP contributes to anoikis resistance of circulating human prostate cancer metastasis precursor cells. AB - Survival in lymph or blood is an essential prerequisite for metastasis of carcinoma cells to distant organs. Recently, we reported isolation and initial biological characterization of circulating metastatic cells in a fluorescent, orthotopic, metastatic nude-mouse model of human prostate cancer. Here we show that the metastatic human prostate carcinoma cells selected for survival in the circulation have increased resistance to anoikis, which is apoptosis induced by cell detachment. Using gene silencing and gene transfer techniques, we show that increased expression of the apoptosis inhibitory protein XIAP contributes to anoikis resistance of the circulating metastatic human prostate carcinoma cells. We also provide initial preclinical data on the antimetastatic efficacy of recently discovered small-molecule antagonists of XIAP. PMID- 15781654 TI - The tumor bed effect: increased metastatic dissemination from hypoxia-induced up regulation of metastasis-promoting gene products. AB - Cancer patients with recurrent local disease after radiation therapy have increased probability of developing regional and distant metastases. The mechanisms behind this observation were studied in the present work by using D-12 and R-18 human melanoma xenografts growing in preirradiated beds in BALB/c-nu/nu mice as preclinical models of recurrent primary tumors in humans. D-12 tumors metastasize to the lungs, whereas R-18 tumors develop lymph node metastases. Based on earlier studies, we hypothesized that metastasis was governed primarily by the proangiogenic factor interleukin-8 (IL-8) in D-12 tumors and by the invasive growth-promoting receptor urokinase-type plasminogen activator receptor (uPAR) in R-18 tumors. Pimonidazole was used as a hypoxia marker, and hypoxia, microvascular hotspots, and the expression of IL-8 and uPAR were studied by immunohistochemistry. The metastatic frequency was significantly higher in tumors in preirradiated beds than in control tumors in unirradiated beds, and it increased with the preirradiation dose. D-12 tumors showed increased fraction of hypoxic cells, increased fraction of IL-8-positive cells, and increased density of microvascular hotspots in preirradiated beds, and R-18 tumors showed increased fraction of hypoxic cells and increased fraction of uPAR-positive cells in preirradiated beds. Strong correlations were found between these parameters and metastatic frequency. IL-8 was up-regulated in hypoxic regions of D-12 tumors, and uPAR was up-regulated in hypoxic regions of R-18 tumors. Daily treatment with anti-IL-8 antibody (D-12) or anti-uPAR antibody (R-18) suppressed metastasis significantly. Our preclinical study suggests that primary tumors recurring after inadequate radiation therapy may show increased metastatic propensity because of increased fraction of hypoxic cells and hypoxia-induced up-regulation of metastasis-promoting gene products. Two possible mechanisms were identified: hypoxia may enhance metastasis by inducing neoangiogenesis facilitating hematogenous spread and by promoting invasive growth facilitating lymphogenous spread. The aggressive behavior of postirradiation local recurrences suggests that they should be subjected to curative treatment as early as possible to prevent further metastatic dissemination. Moreover, the possibility that patients with a high probability of developing local recurrences after radiation therapy may benefit from postirradiation treatment with antiangiogenic and/or anti invasive agents merits clinical investigation. PMID- 15781655 TI - The seco-taxane IDN5390 is able to target class III beta-tubulin and to overcome paclitaxel resistance. AB - A prominent mechanism of drug resistance to taxanes is the overexpression of class III beta-tubulin. The seco-taxane IDN5390 was chosen for its selective activity in paclitaxel-resistant cells with an overexpression of class III beta tubulin. Moreover, the combined treatment paclitaxel/IDN5390 yielded a strong synergism, which was also evident in cell-free tubulin polymerization assays. In the presence of an anti-class III beta-tubulin as a blocking antibody, tubulin polymerization induced by paclitaxel and IDN5390 was enhanced and not affected, respectively, whereas synergism was abolished, thereby indicating that IDN5390 activity is not modulated by class III beta-tubulin levels. Such properties can be explained by taking into consideration the composition of class III beta tubulin paclitaxel binding site; in fact, Ser277 interacting with paclitaxel C group in class I is replaced by an Arginine in class III. IDN5390 that has an open and flexible C ring and an acidic alpha-unsaturated enol-keton moiety better fits with class III beta-tubulin than paclitaxel at the binding site. Taking altogether, these findings indicate that the concomitant treatment IDN5390/paclitaxel is able to successfully target class I and III beta-tubulin and the combined use of two taxanes with diverse spectrum activity against tubulin isotypes could represent a novel approach to overcome paclitaxel resistance. PMID- 15781656 TI - Rapamycin inhibits ezrin-mediated metastatic behavior in a murine model of osteosarcoma. AB - Osteosarcoma is the most frequent primary malignant tumor of bone with a high propensity for metastasis. We have previously showed that ezrin expression is necessary for metastatic behavior in a murine model of osteosarcoma (K7M2). In this study, we found that a mechanism of ezrin-related metastatic behavior is linked to an Akt-dependent mammalian target of rapamycin (mTOR)/p70 ribosomal protein S6 kinase (S6K1)/eukaryotic initiation factor 4E-binding protein 1 (4E BP1) pathway. Suppression of ezrin expression either by antisense transfection or by small interfering RNAs or disruption of ezrin function by transfection of a dominant-negative ezrin-T567A mutant led to decreased expression and decreased phosphorylation of both S6K1 and 4E-BP1. Proteosomal inhibition by MG132 reversed antisense-mediated decrease of S6K1 and 4E-BP1 protein expression, but failed to affect the effect of ezrin on phosphorylation of S6K1 and 4E-BP1. Blockade of the mTOR pathway with rapamycin or its analog, cell cycle inhibitor-779 led to significant inhibition of experimental lung metastasis in vivo. These results suggest that blocking the mTOR/S6K1/4E-BP1 pathway may be an appropriate target for strategies to reduce tumor cell metastasis. PMID- 15781657 TI - Mutant V599EB-Raf regulates growth and vascular development of malignant melanoma tumors. AB - Activating mutations of the B-RAF gene are observed in >60% of human melanomas. Approximately 90% of these mutations occur in the activation segment of the kinase domain as a single-base substitution that converts a valine to glutamic acid at codon 599 (V599E) in exon 15. This mutation causes activation of the kinase as well as downstream effectors of the mitogen-activated protein kinase signaling cascade, leading to melanoma tumor development by an as yet unknown mechanism. In this study, we have identified the role of (V599E)B-Raf in melanoma tumor development by characterizing the mechanism by which this mutant protein promotes melanoma tumorigenesis. Small interfering RNA targeted against B-Raf or a Raf kinase inhibitor (BAY 43-9006) was used to reduce expression and/or activity of (V599E)B-Raf in melanoma tumors. This inhibition led to reduced activity of the mitogen-activated protein kinase-signaling cascade and inhibited tumor development in animals. Targeted reduction of mutant (V599E)B-Raf expression (activity) in melanoma cells before tumor formation inhibited tumorigenesis by reducing the growth potential of melanoma cells. In contrast, reduction of mutant (V599E)B-Raf activity in preexisting tumors prevented further vascular development mediated through decreased vascular endothelial growth factor secretion, subsequently increasing apoptosis in tumors. These effects in combination with reduced proliferative capacity halted growth, but did not shrink the size of preexisting melanoma tumors. Thus, these studies identify the mechanistic underpinnings by which mutant (V599E)B-RAF promotes melanoma development and show the effectiveness of targeting this protein to inhibit melanoma tumor growth. PMID- 15781658 TI - Coadministration of histone deacetylase inhibitors and perifosine synergistically induces apoptosis in human leukemia cells through Akt and ERK1/2 inactivation and the generation of ceramide and reactive oxygen species. AB - Interactions between histone deacetylase inhibitors (HDACIs) and the alkyl lysophospholipid perifosine were examined in human leukemia cells. Coadministration of sodium butyrate, suberoylanilide hydroxamic acid (SAHA), or trichostatin with perifosine synergistically induced mitochondrial dysfunction (cytochrome c and apoptosis-inducing factor release), caspase-3 and -8 activation, apoptosis, and a marked decrease in cell growth in U937 as well as HL 60 and Jurkat leukemia cells. These events were associated with inactivation of extracellular signal-regulated kinase (ERK) 1/2 and Akt, p46 c-jun-NH2-kinase (JNK) activation, and a pronounced increase in generation of ceramide and reactive oxygen species (ROS). They were also associated with up-regulation of Bak and a marked conformational change in Bax accompanied by membrane translocation. Ectopic expression of Bcl-2 delayed but was ultimately ineffective in preventing perifosine/HDACI-mediated apoptosis. Enforced expression of constitutively active mitogen-activated protein kinase kinase (MEK) 1 or myristoylated Akt blocked HDACI/perifosine-mediated ceramide production and cell death, suggesting that MEK/ERK and Akt inactivation play a primary role in these phenomena. However, inhibition of JNK activation (e.g., by the JNK inhibitor SP600125) did not attenuate sodium butyrate/perifosine-induced apoptosis. In addition, the free radical scavenger N-acetyl-L-cysteine attenuated ROS generation and apoptosis mediated by combined treatment. Finally, the acidic sphingomyelinase inhibitor desipramine attenuated HDACI/perifosine-mediated ceramide and ROS production as well as cell death. Together, these findings indicate that coadministration of HDACIs with perifosine in human leukemia cells leads to Akt and MEK/ERK disruption, a marked increase in ceramide and ROS production, and a striking increase in mitochondrial injury and apoptosis. They also raise the possibility that combining these agents may represent a novel antileukemic strategy. PMID- 15781659 TI - Antiangiogenic concentrations of paclitaxel induce an increase in microtubule dynamics in endothelial cells but not in cancer cells. AB - Microtubule-targeted drugs such as paclitaxel exhibit potent antiangiogenic activity at very low concentrations, but the mechanism underlying such an effect remains unknown. To understand the involvement of microtubules in angiogenesis, we analyzed the dynamic instability behavior of microtubules in living endothelial cells [human microvascular endothelial cells (HMEC-1) and human umbilical vascular endothelial cells (HUVEC)] following 4 hours of paclitaxel treatment. Unexpectedly, antiangiogenic concentrations of paclitaxel (0.1-5 nmol/L) strongly increased microtubule overall dynamicity in both HMEC-1 (86 193%) and HUVEC (54-83%). This increase was associated with increased microtubule growth and shortening rates and extents and decreased mean duration of pauses. The enhancement of microtubule dynamics by paclitaxel seemed to be specific to antiangiogenic concentrations and to endothelial cells. Indeed, cytotoxic concentration (100 nmol/L) of paclitaxel suppressed microtubule dynamics by 40% and 54% in HMEC-1 and HUVECs, respectively, as observed for all tested concentrations in A549 tumor cells. After 4 hours of drug incubation, antiangiogenic concentrations of paclitaxel that inhibited endothelial cell proliferation without apoptosis (1-5 nmol/L) induced a slight decrease in anaphase/metaphase ratio, which was more pronounced and associated with increased mitotic index after 24 hours of incubation. Interestingly, the in vitro antiangiogenic effect also occurred at 0.1 nmol/L paclitaxel, a concentration that did not alter mitotic progression and endothelial cell proliferation but was sufficient to increase interphase microtubule dynamics. Altogether, our results show that paclitaxel mediates antiangiogenesis by an increase in microtubule dynamics in living endothelial cells and suggest that the impairment of interphase microtubule functions is responsible for the inhibition of angiogenesis. PMID- 15781660 TI - Mimicry of a cellular low energy status blocks tumor cell anabolism and suppresses the malignant phenotype. AB - Aggressive cancer cells typically show a high rate of energy-consuming anabolic processes driving the synthesis of lipids, proteins, and DNA. Here, we took advantage of the ability of the cell-permeable nucleoside 5-aminoimidazole-4 carboxamide (AICA) riboside to increase the intracellular levels of AICA ribotide, an AMP analogue, mimicking a low energy status of the cell. Treatment of cancer cells with AICA riboside impeded lipogenesis, decreased protein translation, and blocked DNA synthesis. Cells treated with AICA riboside stopped proliferating and lost their invasive properties and their ability to form colonies. When administered in vivo, AICA riboside attenuated the growth of MDA MB-231 tumors in nude mice. These findings point toward a central tie between energy, anabolism, and cancer and suggest that the cellular energy sensing machinery in cancer cells is an exploitable target for cancer prevention and/or therapy. PMID- 15781661 TI - Administration of IFN-alpha enhances the efficacy of a granulocyte macrophage colony stimulating factor-secreting tumor cell vaccine. AB - IFN-alpha is approved for the treatment of multiple cancers. Its pleiotropic properties include inhibition of proliferation and angiogenesis and induction of apoptosis. Type I IFNs also exert immunomodulatory effects, which make it an appropriate candidate to combine with cancer vaccines. The studies reported herein show that 50% of mice reject established B16 tumors following treatment with the combination of a granulocyte macrophage colony-stimulating factor secreting tumor cell vaccine (B16.GM) and subclinical doses of recombinant murine IFN-alpha delivered at the vaccine site. Similarly, 80% of mice treated with the combination reject established B16 tumors when recombinant murine IFN-alpha is given at the challenge site, suggesting that in the latter case its antiproliferative, proapoptotic, and antiangiogenic properties may be involved in controlling tumor growth. In contrast, fewer than 10% of mice reject the tumors when either one is used as a monotherapy. Furthermore, a 30-fold increase in the frequency of melanoma-associated antigen (Trp-2 and gp100) specific T cells was observed in mice treated with the combination when compared with unvaccinated controls. These data show that IFN-alpha combined with a granulocyte macrophage colony-stimulating factor-secreting tumor cell vaccine significantly enhances vaccine potency and may represent a potential new approach for tumor immunotherapy. PMID- 15781662 TI - Increased populations of regulatory T cells in peripheral blood of patients with hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide with a poor prognosis and one for which immunotherapy remains a viable option. Experimental tumor models have shown that regulatory T cells, a functionally unique subset of T cells, can suppress effective antitumor immune responses. This suppression might explain the poor outcome of some of the immunotherapy protocols currently being used. A better understanding of the role of regulatory T cells in HCC is important for design of future immunotherapy-based clinical protocols. We have studied regulatory T cells from 84 patients with HCC and 74 controls, including healthy donors, patients with chronic hepatitis B virus and hepatitis C virus infection and nonviral liver cirrhosis. Regulatory T cells were identified by fluorescence-activated cell sorting using a panel of antibodies and by real time PCR analysis for Foxp3 expression. Functional studies were done to analyze their inhibitory role. Finally, regulatory T cells were analyzed in tumors and ascites from patients with HCC. Patients with HCC have increased numbers of CD4+CD25+ regulatory T cells in their peripheral blood, which express high levels of HLA-DR, GITR, and low or no CD45RA. These cells were anergic toward T-cell receptor stimulation and, when cocultured with activated CD4+CD25- cells, potently suppressed their proliferation and cytokine secretion. There were also high numbers of regulatory T cells in tumor-infiltrating lymphocytes of HCC patients comparable with the increase in their peripheral blood. Our data suggest that the increase in frequency of regulatory T cells might play a role in modulation of the immune response against HCC and could be important in design of immunotherapeutic approaches. PMID- 15781663 TI - Conditional BRAFV600E expression induces DNA synthesis, apoptosis, dedifferentiation, and chromosomal instability in thyroid PCCL3 cells. AB - The activating mutation BRAF(T1796A) is the most prevalent genetic alteration in papillary thyroid carcinomas (PTC). It is associated with advanced PTCs, suggesting that this oncoprotein confers thyroid cancers with more aggressive properties. BRAF(T1796A) is also observed in thyroid micropapillary carcinomas and may thus be an early event in tumor development. To explore its biological consequences, we established doxycycline-inducible BRAF(V600E)-expressing clonal lines derived from well-differentiated rat thyroid PCCL3 cells. Expression of BRAF(V600E) did not induce growth in the absence of thyrotropin despite increasing DNA synthesis, which is likely explained because of a concomitant increase in apoptosis. Thyrotropin-dependent cell growth and DNA synthesis were reduced by BRAF(V600E) because of decreased thyrotropin responsiveness associated with inhibition of thyrotropin receptor gene expression. These results are similar to those obtained following conditional expression of RET/PTC. However, in contrast to RET/PTC, BRAF activation did not impair key activation steps distal to the thyrotropin receptor, such as forskolin-induced adenylyl cyclase activity or cyclic AMP-induced DNA synthesis. We reported previously that acute RET/PTC expression in PCCL3 cells did not induce genomic instability. By contrast, induction of BRAF(V600E) expression increased the frequency of micronuclei by both clastogenic and aneugenic events. These data indicate that BRAF(V600E) expression confers thyroid cells with little growth advantage because of concomitant activation of DNA synthesis and apoptosis. However, in contrast to RET/PTC, BRAF(V600E) may facilitate the acquisition of secondary genetic events through induction of genomic instability, which may account for its aggressive properties. PMID- 15781664 TI - Estrogen enhances whereas tamoxifen retards development of Tsc mouse liver hemangioma: a tumor related to renal angiomyolipoma and pulmonary lymphangioleiomyomatosis. AB - Pulmonary lymphangioleiomyomatosis and abdominal angiomyolipoma are related lesions for which there is no authentic animal model. Both of these proliferative lesions occur in sporadic patients, and at much higher frequency in patients with tuberous sclerosis, which is due to mutations in the TSC1 and TSC2 genes. Tsc1+/- and Tsc2+/- mice frequently develop liver hemangioma. We found that the Tsc mouse liver hemangioma are composed predominantly of endothelial cells but with a smooth muscle component, and express HMB45 antigen, estrogen receptor, and progesterone receptor, similar to lymphangioleiomyomatosis and angiomyolipoma. Estrogen treatment significantly accelerated the development of liver hemangioma in Tsc1+/- female mice, with 91% having liver hemangioma and 55% having severe lesions by 7 months of age. Similarly, an increased frequency and severity of liver hemangiomas was seen in Tsc1+/- males treated with estrogen. In contrast, tamoxifen treatment for 9 months significantly reduced the frequency and severity of hemangiomas in Tsc1+/- female mice. In addition, estrogen treatment significantly increased serum vascular endothelial growth factor levels in Tsc1+/ mice, whereas tamoxifen reduced vascular endothelial growth factor levels. These mouse model observations indicate the importance of estrogen signaling in vivo for the growth of tuberous sclerosis lesions, suggesting the possible benefits of tamoxifen therapy for the treatment of angiomyolipoma and lymphangioleiomyomatosis. PMID- 15781665 TI - Methylenetetrahydrofolate reductase polymorphisms and therapy response in pediatric acute lymphoblastic leukemia. AB - A significant portion of patients treated for pediatric acute lymphoblastic leukemia (ALL) relapse. We hypothesized that common polymorphisms with moderate effect sizes and large attributive risks could explain an important fraction of ALL relapses. Methylenetetrahydrofolate reductase (MTHFR) is central to folate metabolism and has two common functional polymorphisms (C677T and A1298G). Methotrexate (MTX), which interrupts folate metabolism, is a mainstay of pediatric ALL therapy. MTX inhibits the synthesis of dTMP needed for DNA replication by blocking the conversion of 5,10-methylenetetrahydrofolate to 5 methyltetrahydrofolate by MTHFR. We hypothesized that a deactivating MTHFR allele would increase ALL relapse risk by potentially increasing 5,10 methylenetetrahydrofolate and dTMP, enhancing DNA synthesis and thus opposing MTX. To test this hypothesis, we genotyped 520 patients on the Children's Cancer Study Group ALL study, CCG-1891. The MTHFR C677T variant allele was statistically significantly associated with relapse (chi2 = 4.38, P = 0.036). This association remained significant (hazard ratio = 1.82, P = 0.008), controlling for important covariates, and was more predictive of relapse than other predictors, including day 7 bone marrow response. The MTHFR C677T variant allele was not associated with an increased risk of toxicity or infection. The MTHFR A1298G polymorphism was not associated with altered risks of relapse, toxicity, or infection. Haplotype analysis showed six common haplotypes that did not provide additional information predictive for relapse. These data provide evidence that the MTHFR C677T polymorphism is a common genetic variant conferring a moderate relative risk and a high attributable risk for relapse in pediatric ALL patients. PMID- 15781666 TI - 19-Nor-1,25(OH)2D2 (a novel, noncalcemic vitamin D analogue), combined with arsenic trioxide, has potent antitumor activity against myeloid leukemia. AB - Recently, we reported that a novel, noncalcemic vitamin D analogue (19-nor 1,25(OH)2D2; paricalcitol) had anticancer activity. In this study, we explored if paricalcitol enhanced anticancer effects of other clinically useful drugs in vitro against a large variety of cancer cells. Paricalcitol, when combined with As2O3, showed a markedly enhanced antiproliferative effect against acute myeloid leukemia (AML) cells. This combination induced monocytic differentiation of NB-4 acute promyelocytic leukemia (APL) cells and HL-60 AML cells and caused both to undergo apoptosis associated with down-regulation of Bcl-2 and Bcl-x(L). Paricalcitol induced monocytic differentiation of U937 AML cells, which was partially blocked by inducing expression of APL-related PML-retinoic acid receptor alpha (RARalpha) chimeric protein in the U937 cells containing a Zn2+ inducible expression vector coding for this fusion protein (PR9 cells). Exposure to As2O3 decreased levels of PML-RARalpha in PR9 cells, and the combination of paricalcitol and As2O3 enhanced their monocytic differentiation in parallel with the As2O3-mediated decrease of PML-RARalpha. Furthermore, As2O3 increased the transcriptional activity of paricalcitol probably by increasing intracellular levels of paricalcitol by decreasing the function of the mitochondrial enzyme 25 hydroxyvitamin D3-24-hydroxylase, which functions to metabolize the active vitamin D in cells. In summary, the combination of paricalcitol and As2O3 potently decreased growth and induced differentiation and apoptosis of AML cells. This probably occurred by As2O3 decreasing levels of both the repressive PML RARalpha fusion protein and the vitamin D metabolizing protein, 25-hydroxyvitamin D3-24-hydroxylase, resulting in increased activity of paricalcitol. The combination of both of these Food and Drug Administration-approved drugs should be considered for treatment of all-trans retinoic acid-resistant APL patients as well as those with other types of AML. PMID- 15781667 TI - Manganese superoxide dismutase polymorphism, prediagnostic antioxidant status, and risk of clinical significant prostate cancer. AB - Oxidative stress may enhance prostatic carcinogenesis. A polymorphism [valine (V) --> alanine (A)] of manganese superoxide dismutase (MnSOD), the primary antioxidant enzyme in mitochondria, has been recently associated with prostate cancer. We examined the relationship between prostate cancer and the MnSOD polymorphism and its interactions with baseline plasma antioxidant levels (selenium, lycopene, and alpha-tocopherol) and beta-carotene treatment among 567 cases and 764 controls nested in the prospective Physicians' Health Study. We found little overall association between MnSOD polymorphism and prostate cancer risk; however, this polymorphism significantly modified risk of prostate cancer associated with prediagnostic plasma antioxidants (P(interaction) > or = 0.05). Among men with the AA genotype, high selenium level (4th versus 1st quartile) was associated with a relative risk (RR) of 0.3 [95% confidence interval (CI), 0.2 0.7] for total prostate cancer; for clinically aggressive prostate cancer, the RR was 0.2 (95% CI, 0.1-0.5). In contrast, among men with the VV/VA genotype, the RRs were 0.6 (0.4-1.0) and 0.7 (0.4-1.2) for total and clinically aggressive prostate cancer. These patterns were similar for lycopene and alpha-tocopherol and were particularly strong when these antioxidants and selenium were combined; men with the AA genotype had a 10-fold gradient in risk for aggressive prostate cancer across quartiles of antioxidant status. Men with AA genotype who were randomly assigned to beta-carotene treatment (versus placebo) had a RR of 0.6 (95% CI, 0.2-0.9; P(interaction) = 0.03) for fatal prostate cancer, but no significant association was observed in men with the VV/VA genotype. Both endogenous and exogenous antioxidants play an important and interdependent role in preventing clinically significant prostate cancer. PMID- 15781668 TI - Microvascular effects of aldosterone and angiotensin type 2 receptors. PMID- 15781669 TI - Serum uric acid and target organ damage in primary hypertension. AB - The role of serum uric acid as an independent risk factor for cardiovascular and renal morbidity is controversial. A better understanding of its relationship with preclinical organ damage may help clarify the mechanism(s) implicated in the development of early cardiovascular disease. We evaluated the association between uric acid and the presence and degree of target organ damage in 425 (265 males, 160 females) middle-aged, untreated patients with essential hypertension. Left ventricular mass index and carotid intima-media thickness were assessed by ultrasound scan. Albuminuria was measured as the albumin to creatinine ratio in 3 nonconsecutive first morning urine samples. Overall, patients with target organ damage had significantly higher levels of serum uric acid as compared with those without it (presence versus absence of left ventricular hypertrophy, P=0.04; carotid abnormalities, P<0.05; microalbuminuria, P<0.004; and at least 1 versus no organ damage, P<0.03). In women, the occurrence and severity of each target organ damage we examined increased progressively from the lower to the upper serum uric acid tertiles (P<0.01). After adjustment for body mass index, age, creatinine clearance, and high-density lipoprotein cholesterol, each standard deviation increase in serum uric acid entailed a 75% higher risk of having cardiac hypertrophy and a 2-times greater risk of having carotid abnormalities. These results support the role of serum uric acid as an independent, modifiable marker of cardiovascular damage. PMID- 15781670 TI - Low-dose bexarotene and low-dose interferon alfa-2b for adult T-cell leukemia/lymphoma associated with human T-lymphotropic virus 1. PMID- 15781671 TI - Long-term follow-up of patients with early-stage cutaneous T-cell lymphoma who achieved complete remission with psoralen plus UV-A monotherapy. AB - OBJECTIVES: To evaluate long-term outcomes, impact of maintenance therapy and potential curability of patients with mycosis fungoides (MF) treated with psoralen plus UV-A (PUVA) monotherapy. DESIGN: Single-center retrospective cohort analysis. SETTING: Academic referral center for cutaneous lymphoma. PATIENTS: A total of 66 of 104 patients with clinical stages IA to IIA MF who achieved complete remission (CR) after PUVA monotherapy between 1979 and 1995. MAIN OUTCOME MEASURES: Kaplan-Meier actuarial survival and disease-free survival curves were compared between stage IA and IB/IIA cases. Patients were stratified into relapse and nonrelapse groups based on whether their MF relapsed during study follow-up. Baseline characteristics and treatment were compared between these groups. RESULTS: Median follow-up time was 94 months (range, 5-242 months). Thirty-three patients maintained CR for 84 months (range, 5-238 months), and 33 patients experienced relapse with a median disease-free interval of 39 months (range, 2-127 months). There was no significant difference in baseline characteristics between patients in the nonrelapse and relapse groups. Those in the nonrelapse group received a higher cumulative dosage to CR (P = .03) and required longer treatment periods to achieve CR (P = .03). Disease-free survival rates at 5 and 10 years for all patients with stage IA were 56% and 30%, respectively, and for stage IB/IIA, 74% and 50%. Actuarial survival rates at 5, 10, and 15 years were 94%, 82%, and 82%, respectively, in patients with stage IA, and 80%, 69%, and 58% in patients with stage IB/IIA. The overall survival rate for the nonrelapse and relapse groups did not show any statistical difference. One third of the patients developed signs of chronic photodamage and secondary cutaneous malignancies. CONCLUSIONS: Psoralen UV-A is an effective treatment for MF, inducing long-term remissions and perhaps in some cases disease "cure." Thirty percent to 50% of patients remain disease free for 10 years, but late relapses occur. Long-term survival is not affected by relapse status, and the risk of photodamage needs to be measured against the possible benefit of greater disease elimination. PMID- 15781672 TI - Biological effects of bexarotene in cutaneous T-cell lymphoma. AB - OBJECTIVE: To determine the effects of bexarotene on malignant T cells isolated from the peripheral blood of patients with the leukemic variant of cutaneous T cell lymphoma (Sezary syndrome). DESIGN, SETTING, AND PARTICIPANTS: Peripheral blood mononuclear cells from 9 patients with Sezary syndrome and a high burden of circulating malignant T cells (>50% of peripheral blood mononuclear cells) and 6 healthy volunteers underwent evaluation at a university medical center, to test the effects of bexarotene on T cells. MAIN OUTCOME MEASURES: The capacity of bexarotene to induce apoptosis and its effects on T-cell cytokine production from peripheral blood lymphocytes isolated from patients with Sezary syndrome. RESULTS: Bexarotene produced dose-dependent apoptosis of peripheral blood T cells from patients with Sezary syndrome. The T cells from approximately two thirds of patients were consistently sensitive to bexarotene, whereas those from the remaining one third of patients were consistently resistant to the apoptotic effects of bexarotene. Bexarotene inhibited mitogen-induced interleukin 4 production by the peripheral blood cells of patients with Sezary syndrome, and this effect correlated with sensitivity of patients' cells to apoptosis. In contrast to the retinoic acid receptor-specific retinoid, all-trans retinoic acid, bexarotene does not induce the augmentation of interferon gamma production. CONCLUSIONS: Bexarotene induces apoptosis of malignant T cells from patients with Sezary syndrome, but the cells from a proportion of patients are resistant to the apoptotic effects. Interleukin 4 production, which can play a role in the systemic immunosuppression that characterizes advancing Sezary syndrome, may be inhibited by bexarotene. PMID- 15781673 TI - Health-related quality-of-life assessment in patients with cutaneous T-cell lymphoma. AB - OBJECTIVE: To measure and evaluate the health-related quality of life (HRQOL) in patients with cutaneous T-cell lymphoma (CTCL), a visible cutaneous malignancy that may have a profound effect on patients' lives. DESIGN: Monocenter, cross sectional study. SETTING: The Skin Oncology Program, Department of Dermatology, and the Photopheresis Unit of Boston Medical Center. PATIENTS: A total of 22 adult patients with confirmed CTCL. MAIN OUTCOME MEASURES: (1) Evaluation of general oncologic and skin disease-specific HRQOL using, respectively, the Functional Assessment of Cancer Therapy-General (FACT-G) and Skindex-29 profiles; (2) assessment of HRQOL association with disease stage (early stage, IA-IIA; late stage, IIB-IVB). RESULTS: Patients with more advanced CTCL stages reported more effects on general health (FACT-G), particularly in the physical, emotional, and functional domains (P < .05). Patients with early-stage CTCL reported better skin specific HRQOL overall (Skindex-29; P = .002) and for each specific domain than did patients with late-stage disease. The Skindex-29 scales had high internal consistency, and the confirmatory factor structure was similar to that of previous studies. CONCLUSIONS: The HRQOL of patients with CTCL can be evaluated using the Skindex-29 and FACT-G instruments. Patients with more advanced stages of CTCL had lower HRQOL scores. PMID- 15781674 TI - Correlation between serum levels of insulin-like growth factor 1, dehydroepiandrosterone sulfate, and dihydrotestosterone and acne lesion counts in adult women. AB - OBJECTIVES: To determine if insulin-like growth factor 1 (IGF-1) and androgen levels (1) correlate with the presence and severity of acne in adult men and women, and (2) correlate directly with each other and interact in affecting acne. DESIGN: Case-control study and single-center examination of hormone levels in a cohort of volunteers. SETTING: Academic referral center. PATIENTS: Thirty-four subjects (8 women and 8 men with clinical acne, 10 women and 8 men without clinical acne). Clinical acne is defined by a history of persistent acne (acne present on most days for several years), recent acne treatment, and the presence of 10 or more inflammatory acne lesions and 15 or more comedones. INTERVENTIONS: Single visit for serum sampling. MAIN OUTCOME MEASURES: Serum levels of IGF-1 and androgens were determined, adjusted for age, and compared based on the presence or absence of clinical acne using an analysis of covariance. Correlations between hormone levels and acne lesion counts were calculated within each subgroup. Correlations were also calculated between serum levels of IGF-1 and androgens. Further statistical testing was conducted to determine whether IGF-1 or androgens had a greater effect on acne lesion counts. RESULTS: Dehydroepiandrosterone (DHEAS), dihydrotestosterone (DHT), and IGF-1 correlated positively with acne lesion counts in women. Androstenedione and DHEAS correlated with acne lesion counts in men. Although the age-adjusted mean serum levels of IGF-1 were higher in women with clinical acne than in women without clinical acne, this difference did not achieve statistical significance. No difference in IGF-1 level was noted in men based on the presence of clinical acne. In women with clinical acne, IGF-1 correlated with DHT. In men with clinical acne, IGF-1 correlated with DHEAS and androstenedione. In men and women with clinical acne, the effects of androgens on increased acne lesion counts were dependent on the influence of IGF-1. CONCLUSIONS: Increased IGF-1 levels in addition to androgens may influence acne in adult men and women. While IGF-1 appears to have a stronger effect on acne in women, androgens may play a greater role in acne for men. However, in both men and women these hormones are interrelated, possibly owing to reciprocal effects on hormone production. PMID- 15781675 TI - The alopecias associated with vitamin D-dependent rickets type IIA and with hairless gene mutations: a comparative clinical, histologic, and immunohistochemical study. AB - OBJECTIVE: To establish the unique and common clinical and microscopic characteristics of the alopecias associated with vitamin D-dependent rickets (VDDR) type IIA and with hairless gene mutations. DESIGN: A comparative clinical, histologic, and immunohistochemical study of the alopecias in 6 patients with VDDR IIA and 4 patients with atrichia with papular lesions (APL) and/or alopecia universalis congenita (AUC) (hereinafter "APL/AUC"). MAIN OUTCOME MEASURES: Clinical data were gathered from medical records, personal interviews, and physical examinations. Histologic and immunohistochemical studies were performed on 6 scalp punch biopsy specimens from each of the 2 studied groups. RESULTS: The alopecias in VDDR IIA and APL/AUC showed similar clinical, histologic, and immunohistochemical features. The clinical presentation of the VDDR alopecia resembled either the APL phenotype (ie, with papules and milia) or the AUC phenotype (without papules and milia). The main histologic findings included void infundibula; absence of the lower two thirds of the hair follicles, often replaced by vertically oriented irregular epithelial structures or epithelial cysts; irregular epithelial structures, often with small cysts in the middle and lower dermis; and small, medium, and large keratinizing cysts at all levels of the dermis. The larger epithelial cysts in the upper dermis stained positively for cytokeratin (CK) 10, which suggests an infundibular derivation, whereas the remaining irregular epithelial structures and cysts in the middle and lower dermis stained positively most frequently for CK17, CK19, and CD34, which suggests an outer root sheath derivation. CONCLUSIONS: The alopecias associated with VDDR IIA and with hairless gene mutations show striking clinical and microscopic similarities. Disintegration of the lower two thirds of the hair follicles seems to be the underlying defect, and a common pathogenetic pathway might be involved. PMID- 15781676 TI - Mycosis fungoides-type cutaneous T-cell lymphoma and neutrophilic dermatosis. AB - BACKGROUND: Mycosis fungoides (MF) is the most common type of cutaneous T-cell lymphoma (CTCL). Patients with limited patch and/or plaque disease have a normal life expectancy. Neutrophilic dermatosis (ND) may be associated with various hematologic disorders. However, its association with CTCL is exceptional and has been reported only twice with leukemic forms of CTCL. OBSERVATIONS: Three patients with MF developed ND resistant to conventional therapies and responsible for an impaired quality of life due to constitutional symptoms and painful cutaneous lesions. All patients underwent an aggressive treatment course despite their varying initial clinical stages of MF, and all experienced a fatal outcome less than 18 months after the onset of ND. CONCLUSIONS: The association of MF with ND is exceptional and carries a poor prognosis, but the pathophysiologic nature of this association remains unclear. It may involve neutrophil chemoattractant cytokine production by tumor cells. A triggering role of interferon alfa is also possible. PMID- 15781677 TI - Monoclonal T-cell dyscrasia of undetermined significance associated with recalcitrant erythroderma. AB - BACKGROUND: Erythroderma is a diffuse, inflammatory skin reaction that, in rare instances, is associated with hematologic malignancies such as cutaneous T-cell lymphoma (erythrodermic mycosis fungoides) or T-cell leukemia (Sezary syndrome or adult T-cell leukemia/lymphoma). OBSERVATIONS: We screened 30 patients with erythroderma (20 patients with erythroderma of known etiology and 10 patients with idiopathic erythroderma) for the presence of circulating monoclonal T lymphocyte populations using T-cell receptor (TCR)-gamma gene-specific polymerase chain reaction and automated capillary DNA electrophoresis. Moreover, the phenotypic analysis of peripheral blood CD4+ lymphocytes was performed using the following surface markers: CD3, CD7, CD8, CD25, CD26, CD27, CD28, CD29, CD30, CD45RO, CD45RA, CD56, CD134, HLA-DR, TCRalphabeta, TCRgammadelta, and cutaneous lymphocyte antigen (CLA). In 5 patients with idiopathic erythroderma we detected T-cell clones in peripheral blood (in 1 case, associated with the presence of the same clone in the skin) and a 2-fold increase in the proportion of CD3+ CD4+ CD7- CD26- cells. Cell depletion studies indicated that the monoclonal T cells were present within the CD4+ CD7- cell population. Clinically, all patients had chronic, recalcitrant erythroderma but none developed any hematological malignancy during their lifetimes or fulfilled the criteria for cutaneous lymphoma or Sezary syndrome. CONCLUSIONS: A proportion of patients with chronic erythroderma present with the monoclonal expansion of CD4+ CD7- CD26- lymphocytes in their blood. This condition represents a probably benign T-cell dyscrasia, or one of very low malignancy. Alongside monoclonal gammapathy of undetermined significance (MGUS) and monoclonal (B-cell) lymphocytosis of undetermined significance (MLUS), we propose using monoclonal T-cell dyscrasia of undetermined significance (MTUS) to underline a conceptual similarity between this disorder and the more common types of lymphocytic dyscrasia. PMID- 15781678 TI - Imatinib-induced sweet syndrome in a patient with chronic myeloid leukemia. AB - BACKGROUND: Imatinib mesylate has become one of the main chemotherapeutic agents currently used to treat patients with chronic myeloid leukemia (CML). Although cutaneous reactions to this drug have been documented before, this is the first time that Sweet syndrome has been reported with its use. OBSERVATIONS: We report a case of Sweet syndrome secondary to the administration of imatinib to treat CML. On 2 separate occasions, a 53-year-old African American woman with CML developed neutrophilic dermatosis consistent with Sweet syndrome after chemotherapy with imatinib. CONCLUSION: Greater awareness of the adverse effects of imatinib and the characterization of its cutaneous adverse effects will lead to improved surveillance for and treatment of those adverse effects. PMID- 15781679 TI - Effects of UV photographs, photoaging information, and use of sunless tanning lotion on sun protection behaviors. AB - OBJECTIVES: To examine the efficacy of UV photographs and information about photoaging (eg, wrinkles and age spots) for increasing the sun protection intentions and behaviors of young adults and to determine whether any effects of this appearance-based intervention could be enhanced by providing a non-UV alternative for achieving a tan (ie, sunless tanning lotion). DESIGN: Randomized control trial with 1-month follow-up. SETTING: Two universities in Southern California. PARTICIPANTS: A volunteer sample of 146 college students, 91.1% of whom completed the "surprise" 1-month follow-up. INTERVENTION: A UV facial photograph and a brief videotape describing the causes and consequences of photoaging. The study tested the effects of the photoaging information/UV photographic intervention only, the intervention plus use of sunless tanning lotion, and a control condition. MAIN OUTCOME MEASURES: Participants sun protection intentions as assessed immediately after the intervention and sun protection behaviors during the month after the intervention as assessed during a surprise telephone follow-up. RESULTS: The intervention resulted in significantly stronger sun protection intentions (P<.001) and greater sun protection behaviors (P<.05) relative to controls. Furthermore, the group that also used sunless tanning lotion tended to engage in greater sun protection behaviors than the group that received the intervention alone (P<.08). CONCLUSION: The UV photographic intervention holds promise as a cost-effective approach to motivate practices that may ultimately result in health benefits (ie, reduced skin cancer rates). PMID- 15781680 TI - Dyscrasias with "undetermined significance". PMID- 15781682 TI - Painful plaques and nodules on the face, neck, and right wrist of a 51-year-old man. PMID- 15781681 TI - Phacomatosis pigmentovascularis revisited and reclassified. AB - OBJECTIVE: To provide a new comprehensible and practicable classification by use of descriptive terms to distinguish the various types of phacomatosis pigmentovascularis (PPV), which has previously been classified by numbers and letters that are difficult to memorize. STUDY SELECTION: Published case reports on PPV were reassessed. DATA EXTRACTION AND DATA SYNTHESIS: A critical review revealed that only 3 well-established types of PPV so far exist. To eliminate the cumbersome traditional classification by numbering and lettering, the following new terms are proposed: phacomatosis cesioflammea (blue spots [caesius = bluish gray] and nevus flammeus); phacomatosis spilorosea (nevus spilus coexisting with a pale-pink telangiectatic nevus), and phacomatosis cesiomarmorata (blue spots and cutis marmorata telangiectatica congenita). Phacomatosis cesioflammea is identical with the traditional types IIa and IIb; phacomatosis spilorosea corresponds to types IIIa and IIIb; and phacomatosis cesiomarmorata is a descriptive term for type V. A categorical distinction of cases with and without extracutaneous anomalies seems inappropriate. The traditional type I does not exist, and the extremely rare traditional type IV is now included in the group of unclassifiable forms. CONCLUSION: The proposed new classification of PPV by using 3 descriptive terms may be easier to memorize compared with the time-honored grouping of in part not even existing subtypes by numbers and letters. PMID- 15781683 TI - Multiple pigmented nodules. PMID- 15781684 TI - Acral psoriasiform eruption in a man with squamous cell carcinoma of the tonsillar pillar. PMID- 15781685 TI - Black nodular lesions on the foot. PMID- 15781686 TI - Drug-induced eosinophilia and systemic symptoms: hypersensitivity or peripheral T cell lymphoma? PMID- 15781687 TI - Mali acroangiodermatitis in homozygous activated protein C resistance. PMID- 15781688 TI - Multiple eruptive dermatofibromas occurring in a patient with chronic myelogenous leukemia. PMID- 15781689 TI - Subungual melanoma: an insidious erythematous nodule on the nail bed. PMID- 15781690 TI - Dermoscopy of melanocytic neoplasms: subpatterns of melanoma--degrees of malignancy. PMID- 15781691 TI - Saccharomyces cerevisiae histone H2A Ser122 facilitates DNA repair. AB - DNA repair takes place in the context of chromatin. Recently, it has become apparent that proteins that make up and modulate chromatin structure are involved in the detection and repair of DNA lesions. We previously demonstrated that Ser129 in the carboxyl-terminal tail of yeast histone H2A is important for double strand-break responses. By undertaking a systematic site-directed mutagenesis approach, we identified another histone H2A serine residue (Ser122) that is important for survival in the presence of DNA-damaging agents. We show that mutation of this residue does not affect DNA damage-dependent Rad53 phosphorylation or G(2)/M checkpoint responses. Interestingly, we find that yeast lacking H2A S122 are defective in their ability to sporulate. Finally, we demonstrate that H2A S122 provides a function distinct from that of H2A S129. These data demonstrate a role for H2A S122 in facilitating survival in the presence of DNA damage and suggest a potential role in mediating homologous recombination. The distinct roles of H2A S122 and S129 in mediating these responses suggest that chromatin components can provide specialized functions for distinct DNA repair and survival mechanisms and point toward the possibility of a complex DNA damage responsive histone code. PMID- 15781692 TI - Origin and evolution of a chimeric fusion gene in Drosophila subobscura, D. madeirensis and D. guanche. AB - An understanding of the mutational and evolutionary mechanisms underlying the emergence of novel genes is critical to studies of phenotypic and genomic evolution. Here we describe a new example of a recently formed chimeric fusion gene that occurs in Drosophila guanche, D. madeirensis, and D. subobscura. This new gene, which we name Adh-Twain, resulted from an Adh mRNA that retrotransposed into the Gapdh-like gene, CG9010. Adh-Twain is transcribed; its 5' promoters and transcription patterns appear similar to those of CG9010. Population genetic and phylogenetic analyses suggest that the amino acid sequence of Adh-Twain evolved rapidly via directional selection shortly after it arose. Its more recent history, however, is characterized by slower evolution consistent with increasing functional constraints. We present a model for the origin of this new gene and discuss genetic and evolutionary factors affecting the evolution of new genes and functions. PMID- 15781693 TI - Genetical genomics analysis of a yeast segregant population for transcription network inference. AB - Genetic analysis of gene expression in a segregating population, which is expression profiled and genotyped at DNA markers throughout the genome, can reveal regulatory networks of polymorphic genes. We propose an analysis strategy with several steps: (1) genome-wide QTL analysis of all expression profiles to identify eQTL confidence regions, followed by fine mapping of identified eQTL; (2) identification of regulatory candidate genes in each eQTL region; (3) correlation analysis of the expression profiles of the candidates in any eQTL region with the gene affected by the eQTL to reduce the number of candidates; (4) drawing directional links from retained regulatory candidate genes to genes affected by the eQTL and joining links to form networks; and (5) statistical validation and refinement of the inferred network structure. Here, we apply an initial implementation of this strategy to a segregating yeast population. In 65, 7, and 28% of the identified eQTL regions, a single candidate regulatory gene, no gene, or more than one gene was retained in step 3, respectively. Overall, 768 putative regulatory links were retained, 331 of which are the strongest candidate links, as they were retained in the expression correlation analysis and were located within or near an eQTL subregion identified by a multimarker analysis separating multiple linked QTL. One or several biological processes were statistically significantly overrepresented in independent network structures or in highly interconnected subnetworks. Most of the transcription factors found in the inferred network had a putative regulatory link to only one other gene or exhibited cis-regulation. PMID- 15781694 TI - Elephants and human color-blind deuteranopes have identical sets of visual pigments. AB - Being the largest land mammals, elephants have very few natural enemies and are active during both day and night. Compared with those of diurnal and nocturnal animals, the eyes of elephants and other arrhythmic species, such as many ungulates and large carnivores, must function in both the bright light of day and dim light of night. Despite their fundamental importance, the roles of photosensitive molecules, visual pigments, in arrhythmic vision are not well understood. Here we report that elephants (Loxodonta africana and Elephas maximus) use RH1, SWS1, and LWS pigments, which are maximally sensitive to 496, 419, and 552 nm, respectively. These light sensitivities are virtually identical to those of certain "color-blind" people who lack MWS pigments, which are maximally sensitive to 530 nm. During the day, therefore, elephants seem to have the dichromatic color vision of deuteranopes. During the night, however, they are likely to use RH1 and SWS1 pigments and detect light at 420-490 nm. PMID- 15781695 TI - A role for DNA mismatch repair protein Msh2 in error-prone double-strand-break repair in mammalian chromosomes. AB - We examined error-prone nonhomologous end joining (NHEJ) in Msh2-deficient and wild-type Chinese hamster ovary cell lines. A DNA substrate containing a thymidine kinase (tk) gene fused to a neomycin-resistance (neo) gene was stably integrated into cells. The fusion gene was rendered nonfunctional due to a 22-bp oligonucleotide insertion, which included the 18-bp I-SceI endonuclease recognition site, within the tk portion of the fusion gene. A double-strand break (DSB) was induced by transiently expressing the I-SceI endonuclease, and deletions or insertions that restored the tk-neo fusion gene's reading frame were recovered by selecting for G418-resistant colonies. Overall, neither the frequency of recovery of G418-resistant colonies nor the sizes of NHEJ-associated deletions were substantially different for the mutant vs. wild-type cell lines. However, we did observe greater usage of terminal microhomology among NHEJ events recovered from wild-type cells as compared to Msh2 mutants. Our results suggest that Msh2 influences error-prone NHEJ repair at the step of pairing of terminal DNA tails. We also report the recovery from both wild-type and Msh2-deficient cells of an unusual class of NHEJ events associated with multiple deletion intervals, and we discuss a possible mechanism for the generation of these "discontinuous deletions." PMID- 15781696 TI - Bayesian shrinkage estimation of quantitative trait loci parameters. AB - Mapping multiple QTL is a typical problem of variable selection in an oversaturated model because the potential number of QTL can be substantially larger than the sample size. Currently, model selection is still the most effective approach to mapping multiple QTL, although further research is needed. An alternative approach to analyzing an oversaturated model is the shrinkage estimation in which all candidate variables are included in the model but their estimated effects are forced to shrink toward zero. In contrast to the usual shrinkage estimation where all model effects are shrunk by the same factor, we develop a Bayesian method that allows the shrinkage factor to vary across different effects. The new shrinkage method forces marker intervals that contain no QTL to have estimated effects close to zero whereas intervals containing notable QTL have estimated effects subject to virtually no shrinkage. We demonstrate the method using both simulated and real data for QTL mapping. A simulation experiment with 500 backcross (BC) individuals showed that the method can localize closely linked QTL and QTL with effects as small as 1% of the phenotypic variance of the trait. The method was also used to map QTL responsible for wound healing in a family of a (MRL/MPJ x SJL/J) cross with 633 F(2) mice derived from two inbred lines. PMID- 15781697 TI - Characterization of active R2 retrotransposition in the rDNA locus of Drosophila simulans. AB - The rRNA gene (rDNA) loci of all arthropod lineages contain non-LTR retrotransposable elements that have evolved to specifically insert into the 28S rRNA genes. Extensive in vitro experiments have been conducted to investigate the mechanism of R2 retrotransposition but little is known of the insertion frequency or cellular factors that might regulate R2 activity. In this article, isofemale lines obtained from a population of Drosophila simulans were surveyed for recent R2 insertions. Within most lines, all individuals showed the same collection of R2 insertions, providing no evidence for recent R2 activity. However, in a few of the isofemale lines, virtually all individuals differed in their R2 insertion profiles. The descendants of individual pairs of flies from these "active lines" rapidly accumulated new insertions. The frequent insertion of new R2 elements was associated with the elimination of old R2 elements from the rDNA locus. The existence of lines in which R2 retrotransposes frequently and lines in which the elements appear dormant suggests that cellular mechanisms that can regulate the activity of R2 exist. Retrotransposition activity was correlated with the number of full-length R2 elements but not with the size of the rDNA locus or the number of uninserted units. PMID- 15781698 TI - Transmission ratio distortion in intraspecific hybrids of Mimulus guttatus: implications for genomic divergence. AB - We constructed a genetic linkage map between two divergent populations of Mimulus guttatus. We genotyped an F(2) mapping population (N = 539) at 154 AFLP, microsatellite, and gene-based markers. A framework map was constructed consisting of 112 marker loci on 14 linkage groups with a total map length of 1518 cM Kosambi. Nearly half of all markers (48%) exhibited significant transmission ratio distortion (alpha = 0.05). By using a Bayesian multipoint mapping method and visual inspection of significantly distorted markers, we detected 12 transmission ratio distorting loci (TRDL) throughout the genome. The high degree of segregation distortion detected in this intraspecific map indicates substantial genomic divergence that perhaps suggests genomic incompatibilities between these two populations. We compare the pattern of transmission ratio distortion in this map to an interspecific map constructed between M. guttatus and M. nasutus. A similar level of segregation distortion is detected in both maps. Collinear regions between maps are compared to determine if there are shared genetic patterns of non-Mendelian segregation distortion within and among Mimulus species. PMID- 15781699 TI - A gene(s) for all-trans-retinoic acid-induced forelimb defects mapped and confirmed to murine chromosome 11. AB - All-trans-retinoic acid (RA) induces various anatomical limb dysmorphologies in mice dependent on the time of exposure. During early limb development, RA induces forelimb ectrodactyly (digital absence) with varying susceptibilities for different inbred mouse strains; C57BL/6N are highly susceptible while SWV are resistant. To isolate the genetic basis of this defect, a full-genome scan was performed in 406 backcross fetuses of F(1) males to C57BL/6N females. Fetuses were exposed via a maternal injection of 75 mg of RA per kilogram of body weight on gestational day 9.25. The genome-wide analysis revealed significant linkage to a chromosome 11 locus near D11Mit39 with a maximum LOD score of 9.0 and to a chromosome 4 locus near D4Mit170. An epistatic interaction was detected between loci on chromosome 11 (D11Mit39) and chromosome 18 (D18Mit64). Linkage to the chromosome 11 locus (D11Mit39) was confirmed in RA-treated backcross fetuses of F(1) females to C57BL/6N males. Loci associated with bone density/mass in both human and mouse were previously detected in the same region, suggesting a mechanistic linkage with bone homeostasis. The human syntenic region of this locus has been previously linked to Meckel syndrome; the phenotype includes postaxial polydactyly, an ectopic digital defect hypothesized to be induced by a common molecular pathway with ectrodactyly. PMID- 15781700 TI - Quantitative trait locus study design from an information perspective. AB - We examine the efficiency of different genotyping and phenotyping strategies in inbred line crosses from an information perspective. This provides a mathematical framework for the statistical aspects of QTL experimental design, while guiding our intuition. Our central result is a simple formula that quantifies the fraction of missing information of any genotyping strategy in a backcross. It includes the special case of selectively genotyping only the phenotypic extreme individuals. The formula is a function of the square of the phenotype and the uncertainty in our knowledge of the genotypes at a locus. This result is used to answer a variety of questions. First, we examine the cost-information trade-off varying the density of markers and the proportion of extreme phenotypic individuals genotyped. Then we evaluate the information content of selective phenotyping designs and the impact of measurement error in phenotyping. A simple formula quantifies the information content of any combined phenotyping and genotyping design. We extend our results to cover multigenotype crosses, such as the F(2) intercross, and multiple QTL models. We find that when the QTL effect is small, any contrast in a multigenotype cross benefits from selective genotyping in the same manner as in a backcross. The benefit remains in the presence of a second unlinked QTL with small effect (explaining <20% of the variance), but diminishes if the second QTL has a large effect. Software for performing power calculations for backcross and F(2) intercross incorporating selective genotyping and marker spacing is available from http://www.biostat.ucsf.edu/sen. PMID- 15781701 TI - Evolution of salamander life cycles: a major-effect quantitative trait locus contributes to discrete and continuous variation for metamorphic timing. AB - The evolution of alternate modes of development may occur through genetic changes in metamorphic timing. This hypothesis was examined by crossing salamanders that express alternate developmental modes: metamorphosis vs. paedomorphosis. Three strains were used in the crossing design: Ambystoma tigrinum tigrinum (Att; metamorph), wild-caught A. mexicanum (Am; paedomorph), and laboratory Am (paedomorph). Att/Am hybrids were created for each Am strain and then backcrossed to their respective Am line. Previous studies have shown that a dominant allele from Att (met(Att)) and a recessive allele from lab Am (met(lab)) results in metamorphosis in Att/Am hybrids, and met(Att)/met(lab) and met(lab)/met(lab) backcross genotypes are strongly associated with metamorphosis and paedomorphosis, respectively. We typed a molecular marker (contig325) linked to met and found that met(Att)/met(lab) and met(Att)/met(wild) were associated with metamorphosis in 99% of the cases examined. However, the frequency of paedomorphosis was 4.5 times higher for met(lab)/met(lab) than for met(wild)/met(wild). We also found that met(Att)/met(wild) and met(wild)/met(wild) genotypes discriminated distributions of early and late metamorphosing individuals. Two forms of phenotypic variation are contributed by met: continuous variation of metamorphic age and expression of discrete, alternate morphs. We suggest that the evolution of paedomorphosis is associated with genetic changes that delay metamorphic timing in biphasic life cycles. PMID- 15781702 TI - The structure and population genetics of the breakpoints associated with the cosmopolitan chromosomal inversion In(3R)Payne in Drosophila melanogaster. AB - We report here the breakpoint structure and sequences of the Drosophila melanogaster cosmopolitan chromosomal inversion In(3R)P. Combining in situ hybridization to polytene chromosomes and long-range PCR, we have identified and sequenced the distal and proximal breakpoints. The breakpoints are not simple cut and-paste structures; gene fragments and small duplications of DNA are associated with both breaks. The distal breakpoint breaks the tolkin (tok) gene and the proximal breakpoint breaks CG31279 and the tolloid (tld) gene. Functional copies of all three genes are found at the opposite breakpoints. We sequenced a representative sample of standard (St) and In(3R)P karyotypes for a 2-kb portion of the tok gene, as well as the same 2 kb from the pseudogene tok fragment found at the distal breakpoint of In(3R)P chromosomes. The tok gene in St arrangements possesses levels of polymorphism typical of D. melanogaster genes. The functional tok gene associated with In(3R)P shows little polymorphism. Numerous single-base changes, as well as deletions and duplications, are associated with the truncated copy of tok. The overall pattern of polymorphism is consistent with a recent origin of In(3R)P, on the order of Ne generations. The identification of these breakpoint sequences permits a simple PCR-based screen for In(3R)P. PMID- 15781703 TI - The map expansion obtained with recombinant inbred strains and intermated recombinant inbred populations for finite generation designs. AB - The generation of special crosses between different inbred lines such as recombinant inbred strains (RIS) and intermated recombinant inbred populations (IRIP) is being used to improve the power of QTL detection techniques, in particular fine mapping. These approaches acknowledge the fact that recombination of linked loci increases with every generation, caused by the accumulation of crossovers appearing between the loci at each meiosis. This leads to an expansion of the map distance between the loci. While the amount of the map expansion of RIS and IRIP is known for infinite inbred generations, it is not known for finite numbers of generations. This gap was closed here. Since the recursive evaluation of the map expansion factors turned out to be complex, a useful approximation was derived. PMID- 15781704 TI - Intron size and exon evolution in Drosophila. AB - We have found a negative correlation between evolutionary rate at the protein level (as measured by d(N)) and intron size in Drosophila. Although such a relation is expected if introns reduce Hill-Robertson interference within genes, it seems more likely to be explained by the higher abundance of cis-regulatory elements in introns (especially first introns) in genes under strong selective constraints. PMID- 15781705 TI - A unified model of codon reassignment in alternative genetic codes. AB - Many modified genetic codes are found in specific genomes in which one or more codons have been reassigned to a different amino acid from that in the canonical code. We present a new framework for codon reassignment that incorporates two previously proposed mechanisms (codon disappearance and ambiguous intermediate) and introduces two further mechanisms (unassigned codon and compensatory change). Our theory is based on the observation that reassignment involves a gain and a loss. The loss could be the deletion or loss of function of a tRNA or release factor. The gain could be the gain of a new type of tRNA or the gain of function of an existing tRNA due to mutation or base modification. The four mechanisms are distinguished by whether the codon disappears from the genome during the reassignment and by the order of the gain and loss events. We present simulations of the gain-loss model showing that all four mechanisms can occur within the same framework as the parameters are varied. We investigate the way the frequencies of the mechanisms are influenced by selection strengths, the number of codons undergoing reassignment, directional mutation pressure, and selection for reduced genome size. PMID- 15781706 TI - Self-fertilization and the evolution of recombination. AB - In this article, we study the effect of self-fertilization on the evolution of a modifier allele that alters the recombination rate between two selected loci. We consider two different life cycles: under gametophytic selfing, a given proportion of fertilizations involves gametes produced by the same haploid individual, while under sporophytic selfing, a proportion of fertilizations involves gametes produced by the same diploid individual. Under both life cycles, we derive approximations for the change in frequency of the recombination modifier when selection is weak relative to recombination, so that the population reaches a state of quasi-linkage equilibrium. We find that gametophytic selfing increases the range of epistasis under which increased recombination is favored; however, this effect is substantial only for high selfing rates. Moreover, gametophytic selfing affects the relative influence of different components of epistasis (additive x additive, additive x dominance, dominance x dominance) on the evolution of the modifier. Sporophytic selfing has much stronger effects: even a small selfing rate greatly increases the parameter range under which recombination is favored, when there is negative dominance x dominance epistasis. This effect is due to the fact that selfing generates a correlation in homozygosity at linked loci, which is reduced by recombination. PMID- 15781707 TI - Quantitative trait loci that control dengue-2 virus dissemination in the mosquito Aedes aegypti. AB - The mosquito Aedes aegypti is the most important vector of yellow fever and dengue fever flaviviruses. Ae. aegypti eradication campaigns have not been sustainable and there are no effective vaccines for dengue viruses. Alternative control strategies may depend upon identification of mosquito genes that condition flavivirus susceptibility and may ultimately provide clues for interrupting transmission. Quantitative trait loci affecting the ability of Ae. aegypti to develop a dengue-2 infection in the midgut have been mapped previously. Herein we report on QTL that determine whether mosquitoes with a dengue-2-infected gut can then disseminate the virus to other tissues. A strain selected for high rates of dengue-2 dissemination was crossed to a strain selected for low dissemination rates. QTL were mapped in the F(2) and again in an F(5) advanced intercross line. QTL were detected at 31 cM on chromosome I, at 32 cM on chromosome II, and between 44 and 52 cM on chromosome III. Alleles at these QTL were additive or dominant in determining rates of dengue-2 dissemination and accounted for approximately 45% of the phenotypic variance. The locations of dengue-2 midgut infection and dissemination QTL correspond to those found in earlier studies. PMID- 15781708 TI - Natural allelic variation in the temperature-compensation mechanisms of the Arabidopsis thaliana circadian clock. AB - Temperature compensation is a defining feature of circadian oscillators, yet no components contributing to the phenomenon have been identified in plants. We tested 27 accessions of Arabidopsis thaliana for circadian leaf movement at a range of constant temperatures. The accessions showed varying patterns of temperature compensation, but no clear associations to the geographic origin of the accessions could be made. Quantitative trait loci (QTL) were mapped for period and amplitude of leaf movement in the Columbia by Landsberg erecta (CoL) and Cape Verde Islands by Landsberg erecta (CvL) recombinant inbred lines (RILs) at 12 degrees , 22 degrees , and 27 degrees . Six CvL and three CoL QTL were located for circadian period. All of the period QTL were temperature specific, suggesting that they may be involved in temperature compensation. The flowering time gene GIGANTEA and F-box protein ZEITLUPE were identified as strong candidates for two of the QTL on the basis of mapping in near isogenic lines (NILs) and sequence comparison. The identity of these and other candidates suggests that temperature compensation is not wholly determined by the intrinsic properties of the central clock proteins in Arabidopsis, but rather by other genes that act in trans to alter the regulation of these core proteins. PMID- 15781710 TI - Improving quantitative trait loci mapping resolution in experimental crosses by the use of genotypically selected samples. AB - One of the key factors contributing to the success of a quantitative trait locus (QTL) mapping experiment is the precision with which QTL positions can be estimated. We show, using simulations, that QTL mapping precision for an experimental cross can be increased by the use of a genotypically selected sample of individuals rather than an unselected sample of the same size. Selection is performed using a previously described method that optimizes the complementarity of the crossover sites within the sample. Although the increase in precision is accompanied by a decrease in QTL detection power at markers distant from QTL, only a modest increase in marker density is needed to obtain equivalent power over the whole map. Selected samples also show a slight reduction in the number of false-positive QTL. We find that two features of selected samples independently contribute to these effects: an increase in the number of crossover sites and increased evenness in crossover spacing. We provide an empirical formula for crossover enrichment in selected samples that is useful in experimental design and data analysis. For QTL studies in which the phenotyping is more of a limiting factor than the generation of individuals and the scoring of genotypes, selective sampling is an attractive strategy for increasing genome wide QTL map resolution. PMID- 15781709 TI - The Ku protein complex is involved in length regulation of Drosophila telomeres. AB - Chromosome ends in Drosophila melanogaster can be elongated either by terminal attachment of the telomere-specific retrotransposons HeT-A and TART or by terminal gene conversion. Here we show that a decrease in Ku70 or Ku80 gene dosage causes a sharp increase in the frequency of HeT-A and TART attachments to a broken chromosome end and in terminal DNA elongation by gene conversion. Loss of Ku80 has more pronounced effects than loss of Ku70. However, lower Ku70 concentration reduces the stability of terminally deficient chromosomes. Our results suggest a role of the end-binding Ku complex in the accessibility and length regulation of Drosophila telomeres. PMID- 15781711 TI - Using pool noodles to teach mitosis and meiosis. AB - Although mitosis and meiosis are fundamental to understanding genetics, students often find them difficult to learn. We suggest using common "pool noodles" as teaching aids to represent chromatids in classroom demonstrations. Students use these noodles to demonstrate the processes of synapsis, segregation, and recombination. Student feedback has been overwhelmingly positive. PMID- 15781712 TI - Mutations of a redundant alpha-tubulin gene affect Caenorhabditis elegans early embryonic cleavage via MEI-1/katanin-dependent and -independent pathways. AB - The C. elegans zygote supports both meiosis and mitosis within a common cytoplasm. The meiotic spindle is small and is located anteriorly, whereas the first mitotic spindle fills the zygote. The C. elegans microtubule-severing complex, katanin, is encoded by the mei-1 and mei-2 genes and is solely required for oocyte meiotic spindle formation; ectopic mitotic katanin activity disrupts mitotic spindles. Here we characterize two mutations that rescue the lethality caused by ectopic MEI-1/MEI-2. Both mutations are gain-of-function alleles of tba 2 alpha-tubulin. These tba-2 alleles do not prevent MEI-1/MEI-2 microtubule localization but do interfere with its activity. TBA-1 and TBA-2 are redundant for viability, but when katanin activity is limiting, TBA-2 is preferred over TBA 1 by katanin. This is similar to what we previously reported for the beta tubulins. Removing both preferred alpha- and beta-isoforms results in normal development, suggesting that the katanin isoform preferences are not absolute. We conclude that while the C. elegans embryo expresses redundant alpha- and beta tubulin isoforms, they nevertheless have subtle functional specializations. Finally, we identified a dominant tba-2 allele that disrupts both meiotic and mitotic spindle formation independently of MEI-1/MEI-2 activity. Genetic studies suggest that this tba-2 mutation has a "poisonous" effect on microtubule function. PMID- 15781713 TI - The origin of subfunctions and modular gene regulation. AB - Evolutionary explanations for the origin of modularity in genetic and developmental pathways generally assume that modularity confers a selective advantage. However, our results suggest that even in the absence of any direct selective advantage, genotypic modularity may increase through the formation of new subfunctions under near-neutral processes. Two subfunctions may be formed from a single ancestral subfunction by the process of fission. Subfunction fission occurs when multiple functions under unified genetic control become subdivided into more restricted functions under independent genetic control. Provided that population size is sufficiently small, random genetic drift and mutation can conspire to produce changes in the number of subfunctions in the genome of a species without necessarily altering the phenotype. Extensive genotypic modularity may then accrue in a near-neutral fashion in permissive population-genetic environments, potentially opening novel pathways to morphological evolution. Many aspects of gene complexity in multicellular eukaryotes may have arisen passively as population size reductions accompanied increases in organism size, with the adaptive exploitation of such complexity occurring secondarily. PMID- 15781714 TI - Stochastic models for horizontal gene transfer: taking a random walk through tree space. AB - Horizontal gene transfer (HGT) plays a critical role in evolution across all domains of life with important biological and medical implications. I propose a simple class of stochastic models to examine HGT using multiple orthologous gene alignments. The models function in a hierarchical phylogenetic framework. The top level of the hierarchy is based on a random walk process in "tree space" that allows for the development of a joint probabilistic distribution over multiple gene trees and an unknown, but estimable species tree. I consider two general forms of random walks. The first form is derived from the subtree prune and regraft (SPR) operator that mirrors the observed effects that HGT has on inferred trees. The second form is based on walks over complete graphs and offers numerically tractable solutions for an increasing number of taxa. The bottom level of the hierarchy utilizes standard phylogenetic models to reconstruct gene trees given multiple gene alignments conditional on the random walk process. I develop a well-mixing Markov chain Monte Carlo algorithm to fit the models in a Bayesian framework. I demonstrate the flexibility of these stochastic models to test competing ideas about HGT by examining the complexity hypothesis. Using 144 orthologous gene alignments from six prokaryotes previously collected and analyzed, Bayesian model selection finds support for (1) the SPR model over the alternative form, (2) the 16S rRNA reconstruction as the most likely species tree, and (3) increased HGT of operational genes compared to informational genes. PMID- 15781715 TI - Experimental designs for reliable detection of linkage disequilibrium in unstructured random population association studies. AB - A method is given for design of experiments to detect associations (linkage disequilibrium) in a random population between a marker and a quantitative trait locus (QTL), or gene, with a given strength of evidence, as defined by the Bayes factor. Using a version of the Bayes factor that can be linked to the value of an F-statistic with an existing deterministic power calculation makes it possible to rapidly evaluate a comprehensive range of scenarios, demonstrating the feasibility, or otherwise, of detecting genes of small effect. The Bayes factor is advocated for use in determining optimal strategies for selecting candidate genes for further testing or applications. The prospects for fine-scale mapping of QTL are reevaluated in this framework. We show that large sample sizes are needed to detect small-effect genes with a respectable-sized Bayes factor, and to have good power to detect a QTL allele at low frequency it is necessary to have a marker with similar allele frequency near the gene. PMID- 15781716 TI - Straightening the crooked middle third of the nose: using porous polyethylene extended spreader grafts. AB - OBJECTIVE: To evaluate a surgical technique used to straighten and support the crooked middle third of the nose. DESIGN: Retrospective review of 41 patients with a markedly crooked nose who underwent correction using extended spreader grafts made from high-density porous polyethylene. Follow-up ranged from 6 months to 2 years. RESULTS: Every patient had substantial improvement in the straightening of the middle third of the crooked nose and the airway in one procedure. There were no cases of extrusion or infection. CONCLUSIONS: The extended spreader graft technique introduces structural reinforcement to permit straightening of the middle third of the nose. It is safe, effective, and reliable. The graft material provides increased strength against further trauma or the long-term forces of scar contracture. This technique has compelling benefits compared with all other techniques used to straighten the severely crooked middle third of the nose. PMID- 15781718 TI - The long-term effects of alar base reduction. AB - OBJECTIVE: To statistically analyze the long-term results of alar base reduction after rhinoplasty. METHODS: Among a consecutive series of 100 rhinoplasty cases, 19 patients required alar base reduction. The mean (SD) follow-up time was 11 (9) months (range, 2 months to 3 years). Using preoperative and postoperative photographs, comparisons were made of the change in the base width (width of base between left and right alar-facial junctions), flare width (width on base view between points of widest alar flare), base height (distance from base to nasal tip on base view), nostril height (distance from base to anterior edge of nostril), and vertical flare (vertical distance from base to the widest alar flare). Notching at the nasal sill was recorded as none, minimal, mild, moderate, and severe. RESULTS: Changes in vertical flare (P<.05) and nostril height (P<.05) were the only significant differences seen in the patients who required alar reduction. No significant change was seen in base width (P=.92), flare width (P=.41), or base height (P=.22). No notching was noted. CONCLUSIONS: It would have been preferable to study patients undergoing alar reduction without concomitant rhinoplasty procedures, but this approach is not practical. To our knowledge, the present study represents the most extensive attempt in the literature to characterize and quantify the postoperative effects of alar base reduction. PMID- 15781717 TI - A simplified approach to alar base reduction: a review of 124 patients over 20 years. AB - OBJECTIVES: To simplify the approach and diagnosis of alar base reduction, suggest a treatment algorithm, and evaluate the long-term outcomes of 3 different techniques used separately or in conjunction with one another. DESIGN: Retrospective review of 124 patients seen in a private practice by a single surgeon. Patients ranged in age from 15 to 59 years (mean age, 30.4 years). Patients were undergoing primary (83.9%) or revision (16.1%) procedures. RESULTS: Of the 124 patients undergoing alar base reduction, 31 (25%) were male and 93 (75%) were female. Average follow-up was 2 years. All patients underwent wedge excision, and for 64 patients (51.6%), this was the only technique used on the alar base. Alar wedge and nasal sill excisions were performed in 21 patients (16.9%); 19 (15.3%) underwent alar wedge excision with V-Y advancement, and 20 (16.1%) underwent alar wedge excision, nasal sill excision, and V-Y advancement. Thirty-one patients (25.0%) received dermabrasion for notable postoperative incision scars. CONCLUSIONS: The data represent the senior author's outcomes of alar base reductions over the past 20 years. The 3 techniques we describe have been effective when used alone or in combination in reducing alar flare and in narrowing the nasal base. Patients should be counseled that dermabrasion of the wedge excision areas in the alar-facial groove may be necessary to diminish visible scars. PMID- 15781719 TI - Alar base reduction. PMID- 15781720 TI - Reconstruction after resection of sphenoid wing meningiomas. AB - OBJECTIVE: To review our experience of reconstructing the lateral and superior orbital walls after resection of sphenoid wing meningiomas. We will review the presentation and complications, examine the aesthetic results postoperatively, and compare preoperative and postoperative computed tomographic scans. To our knowledge, a comparative analysis of preoperative defect and postoperative reconstruction has not been performed. METHODS: We conducted a retrospective analysis, with a minimum of 5 months and a maximum of 9 years of follow-up in an academic multidisciplinary skull base center. Twenty-two patients were treated for sphenoid wing meningiomas by resection and reconstruction with split calvarial bone graft and, for more than half of the patients, also with free abdominal fat graft. The main outcome measures were aesthetic evaluation of patients and analysis of tumor control using computed tomographic scans, survival, and complications. RESULTS: A total of 24 resections were performed on 22 patients. The average follow-up was 14.6 months. All patients had meningiomas with similar preoperative presentations, and for 21 of the 22 patients aesthetic reconstruction resulted in the near symmetry of the 2 sides. All patients are currently alive, those who underwent complete resection are without recurrence, and 15 (68.2%) did not incur complications. One patient experienced a worsening of temporal wasting following radiation therapy. CONCLUSION: Reconstruction of the defect with split calvarial bone and free abdominal fat grafts affords the patient excellent aesthetic results as well as good symmetry, as demonstrated by a postoperative computed tomographic scan. PMID- 15781721 TI - Enzymatic recontouring of auricular cartilage in a rabbit model. AB - OBJECTIVE: To evaluate the effectiveness of contouring auricular cartilage in a rabbit model using biologically active enzymes injected subcutaneously. METHODS: The first phase determined the most effective volume and concentration required to affect the cartilage. To accomplish this task, we used ex vivo rabbit ears from a slaughterhouse. In the second phase, we injected 1 mL of hyaluronidase (150 U per milliliter of isotonic sodium chloride solution [saline]), elastase (1 mg per milliliter of saline), or saline into the ears of live rabbits. The study took place at the Madigan Army Medical Center (Tacoma, Wash), and included 10 animals. In each rabbit, we injected the test compound in one ear and saline in the other ear (control). We injected hyaluronidase in 5 ears and elastase in 5 ears. After injection, the ears were contoured and splinted for 4 weeks. In the third phase, we changed the injection pathway in 5 animals. RESULTS: At 4 weeks, 4 (80%) of the 5 ears injected with hyaluronidase showed full response and 1 (20%) had a partial response. Of the 5 ears injected with elastase, 4 (80%) showed a full response while 1 (20%) demonstrated a partial response. There was a response in all 10 of the ears injected with a test compound. Of the 10 control ears, 3 (30%) showed a partial response. At 6 weeks, approximately 6 (30%) of the ears had maintained contour demonstrating a full response. The difference between the test ears and the control ears was statistically significant (P = .006). Compared with the control ears, the results were statistically significant for elastase (P = .004) and hyaluronidase (P = .02). Overall, both agents demonstrated a subjective and objective response compared with control ears. CONCLUSION: This study demonstrates that bioactive enzymes and splinting can be effective in correcting ear deformities in a rabbit model. PMID- 15781722 TI - Aesthetic proportions of the nasal aperture in 3 different racial groups of men. AB - OBJECTIVE: To define baseline aesthetic dimensions of the nasal aperture in 3 different racial groups. METHODS: Healthy volunteers from 3 different racial groups (15 white, 15 Chinese, and 15 Indian men) were enrolled in the study at the Department of Otolaryngology, University of Dundee, Dundee, Scotland. Those with a history of nasal or facial surgery or trauma were excluded from the study. Images were obtained and stored in a digital format. The dimensions of nasal aperture were defined by the length of the columella at the narrowest point, the width of the columella at the narrowest point, the length of the nasal aperture at the maximum length, the width of the nasal aperture at the maximum width, and the width of the alar cartilage base. RESULTS: There was no significant difference in the length or the width of the columella for the 3 racial groups. There was no significant difference in the length of the nasal aperture between the Chinese and the white groups. The nasal aperture was longer in the Indian group compared with the other 2 groups (P<.002). The nasal aperture at the maximum width was narrower in the Chinese group compared with the other groups (P<.002); there was no significant difference between the white and Indian groups. The nasal alar width was slightly narrower at the alar base in the Chinese group compared with other racial groups (P<.001). CONCLUSIONS: The aesthetic dimensions of the nasal aperture differ between racial groups. The nasal aperture and the alar base were narrower in the Chinese group, and the nasal aperture was longer in the Indian group. The aesthetic surgeon should ideally have an understanding of these ethnic variations. PMID- 15781723 TI - An electrophysiologic model for functional assessment of effects of neurotrophic factors on facial nerve reinnervation. AB - OBJECTIVES: To establish a sound objective model for assessing the effects of neurotrophic factors on facial nerve function after injury and to compare the effects of brain-derived neurotrophic factor (BDNF) with its neutralizing antibody on facial nerve function after injury. DESIGN: Prospective electrophysiologic analysis of recovery of function 4 weeks after axotomy involving facial nerve transection and primary end-to-end reanastomosis in adult rats and blind comparison with randomized intramuscular injection of either BDNF, monoclonal antibody to BDNF in neutralizing concentration, or control solution. RESULTS: There were no statistically significant differences between groups in latencies, duration, amplitude, area, or conduction velocity before axotomy, and recorded conduction velocities were consistent with previously reported values, which suggests that the recordings were reliable and reproducible. After transection, there was a mean increase in latency 1 and decreases in latency 2, integrated average area, muscle action potential duration, amplitude, and conduction velocity for all 3 groups. When the groups were compared after transection, the anti-BDNF group showed a significant decrease in conduction velocity and muscle action potential duration (Kruskal-Wallis P = .01 and P = .008, respectively) compared with the other groups. There were no statistically significant differences in latencies, amplitude, or area among the groups. CONCLUSIONS: We have established an electrophysiologic model for objective assessment of facial nerve function in the rat. Future studies should combine functional electrophysiologic assessment and histologic examination to provide a more robust model for studying the effects of neurotrophic factors on facial nerve reinnervation and synkinesis. PMID- 15781724 TI - Contralateral platysma suspension: an adjunct to rhytidectomy. AB - As we age, a number of factors distort the contour of the cervicomental angle. Submental adipose deposits, anterior laxity of the platysma muscle, and relaxation of cervical skin contribute to this undesirable result. Comprehensive rhytidectomy will frequently involve enhancement of the cervicomental angle as an adjunctive procedure. Facial aesthetic surgeons have devised numerous techniques to deal with the aging neck. Commonly, the approach combines submental adipose tissue excision with posterosuperior-based support, either by preauricular or postauricular incision. Herein, we describe a technique incorporating standard submental suction lipectomy with a novel method of contralateral midline platysma suspension that creates a dynamic support system involving both rigid suture based and flexible muscle-based components. This technique delivers quality aesthetic results and superior postoperative patient satisfaction. PMID- 15781725 TI - Diagnosis and surgical management of the caudal excess nasal deformity. AB - Surgical correction of the overgrown columella-labial junction remains a controversial and neglected aspect of cosmetic rhinoplasty. Failure to reduce an obtuse nasolabial angle resulting from overgrowth of the septum and/or nasal spine may adversely affect the otherwise satisfactory cosmetic result. I review the historical considerations regarding caudal overgrowth and discuss its relevance to contemporary rhinoplasty. Case studies are presented to illustrate surgical treatment options for various overgrowth deformities. PMID- 15781726 TI - Color-specific enhancement of digital photographs for identification of the extent of cutaneous malignancy. AB - OBJECTIVES: To introduce the concept of active digital imaging to the literature and to support further investigation by showing the utility of photochromatography in the identification of cutaneous cancer margins METHODS: Digital color images of 10 cutaneous basal cell carcinomas were digitally enhanced to highlight color change in and around each lesion. After the most intense area of tumor coloration was located and sampled, the color was digitally changed to a sharply contrasting color across the entire region, thereby highlighting abnormal areas not readily seen by the unaided eye. The enhanced areas of predicted tumor extent were compared with digital images of the resulting defect after treatment with Mohs micrographic surgery. RESULTS: The extent of tumor was predicted with good accuracy in 5 cases and with fair accuracy in 2 cases and was not determinable in the remaining 2 cases. In no case did photochromatography overestimate the extent of the lesion. CONCLUSIONS: Digital highlighting of color change not readily seen by the unaided eye (photochromatography) can improve identification and localization of cutaneous tumor. Our findings justify further investigation into algorithms for photographic color detection and enhancement in the evaluation of tissue change. PMID- 15781727 TI - Interest in facial plastic and reconstructive surgery among otorhinolaryngologists: a survey in The Netherlands. AB - OBJECTIVE: To assess the interest of Dutch otorhinolaryngologists in facial plastic and reconstructive surgery (FPRS). METHODS: We conducted a 22-question survey among otorhinolaryngology physicians and residents concerning their experience with and interest in FPRS. The response rate was 71% (335/475; 275 physicians and 60 residents). RESULTS: Most respondents associated FPRS with rhinoplasty, otoplasty, and the reconstruction of skin cancer defects. Of the physicians, 81% said that 1% to 33% of their practice involves FPRS; 62% were satisfied with this percentage, whereas 36% would like it to be higher. Approximately 70% of physicians regarded their training in FPRS as insufficient, although most (70%) had taken supplementary courses. Moreover, 73% of the otorhinolaryngology physicians and 72% of all respondents said that FPRS should be taught during and after residency, with a preference for hands-on courses. Finally, 84% of all respondents thought that FPRS should be part of the field of otorhinolaryngology, whereas 48% thought that it should become a subspecialty. CONCLUSION: There is interest in integrating FPRS training into the Dutch otorhinolaryngology residency program, as it is in the United States. PMID- 15781728 TI - Perceptions regarding facial plastic surgery among Dutch otolaryngologists. PMID- 15781729 TI - Frans Hals' Willem Coymans. PMID- 15781730 TI - Adenovirus-mediated expression of beta-adrenergic receptor kinase C-terminus reduces intimal hyperplasia and luminal stenosis of arteriovenous polytetrafluoroethylene grafts in pigs. AB - BACKGROUND: Hemodialysis vascular access dysfunction is the single most important cause of morbidity in kidney hemodialysis patients. Failure of an arteriovenous polytetrafluoroethylene (PTFE) graft, the most common form of hemodialysis access, is primarily due to intimal hyperplasia and thrombosis at the venous anastomosis. METHODS AND RESULTS: This study was aimed at evaluating the efficacy and safety of an adenoviral vector (Ad2/betaARKct) encoding the carboxyl terminus of beta-adrenergic receptor kinase (betaARKct) in a pig model of arteriovenous PTFE graft failure. Transduction of the external jugular vein with Ad2/betaARKct (5E9, 5E10, or 5E11 particles per vein) did not result in systemic toxicity, as measured by clinical and pathological assessments. Ad2/betaARKct significantly reduced neointimal hyperplasia in the graft/vein anastomosis. It also improved the graft patency rate and angiographic score, as measured histologically and angiographically, compared with vehicle or empty viral vector controls. CONCLUSIONS: Our results suggest that local administration of adenoviral vectors encoding betaARKct into the jugular vein represents a viable strategy to treat AV graft hemodialysis vascular access failure. PMID- 15781731 TI - The use of nonsteroidal anti-inflammatory drugs (NSAIDs): a science advisory from the American Heart Association. PMID- 15781732 TI - Long-term serotonin administration induces heart valve disease in rats. AB - BACKGROUND: The purpose of this study was to investigate whether rats dosed with serotonin develop changes similar to those seen in human carcinoid heart disease. METHODS AND RESULTS: Ten Sprague-Dawley rats were given serotonin injections subcutaneously once daily for 3 months; controls were given saline. A long lasting hyperserotoninemia with a >10-fold increase in both platelet-poor plasma and dialysate from the femoral muscles appeared. The animals developed clinical signs such as flushing and loose stools. After 3 months, 6 of 10 rats given serotonin had pathological echocardiographs. Two animals had a combination of aortic and pulmonary valve insufficiency, 1 had isolated aortic valve insufficiency, and 3 had isolated pulmonary valve insufficiency. Histopathological examination revealed shortened and thickened aortic cusps and carcinoidlike plaques characterized by a collection of myofibroblasts within an extracellular matrix of collagen ground substance. Immunostaining for Ki-67 demonstrated an increased number of proliferating subendocardial cells. In the control group, no pathological changes were seen. With the use of reverse transcription polymerase chain reaction, normal rat aortic cusps were shown to express mRNA for serotonin receptors 5-HT1A, 5-HT2A, and 5-HT2B and the serotonin transporter 5-HTT. CONCLUSIONS: For the first time, long-term serotonin administration was performed in rats. Morphological and echocardiographic changes similar to those seen in human carcinoid heart disease developed. This study demonstrates that serotonin most likely is involved in the pathogenesis of carcinoid heart disease. PMID- 15781734 TI - Abnormalities of Doppler measures of diastolic function in the healthy elderly are not related to alterations of left atrial pressure. AB - BACKGROUND: Normal healthy aging results in changes in Doppler indexes of diastolic function. One widely accepted explanation for these alterations that is based on animal data is an age-associated impairment of myocardial relaxation resulting from abnormal myocyte calcium handling. However, an alternative theory based on altered left atrial compliance with aging has been proposed in which early left atrial pressure could be low in the aged heart but rise rapidly during atrial diastole, resulting in a higher late atrial pressure and thus a normal mean pressure. We sought to explore this issue directly by performing a detailed analysis of the pulmonary capillary wedge pressure waveform obtained by right heart catheterization during 5 different loading conditions. METHODS AND RESULTS: Twelve healthy elderly sedentary subjects (mean age, 69.8+/-3 years) were recruited for the seniors group. An additional 12 young sedentary subjects (mean age, 35+/-8 years) made up the young group. All subjects were rigorously screened for comorbidities. All subjects underwent transthoracic echocardiography to determine Doppler variables of early and late transmitral filling (E and A velocities) and isovolumetric relaxation time. Each subject also underwent pulmonary artery catheterization with measurement of pulmonary capillary wedge pressure waveform during 5 different loading conditions: baseline, lower-body negative pressures of -15 and -30 mm Hg, and rapid saline infusions of 10 to 15 and 20 to 30 mL/kg. Pressure was measured at 6 points of the waveform: point 1, peak of the atrial contraction (a wave); point 2, the left atrial pressure during the start of ventricular systole; point 3, peak of atrial filling (v wave); point 4, earliest left atrial pressure during ventricular filling; and the line between points 5 and 6, pressure during diastasis. Aging resulted in a decrease in the E/A ratio (P<0.001) and a prolongation of the isovolumetric relaxation time (P<0.001) as assessed by echocardiography, but there was no effect of age on pulmonary capillary wedge pressure at any point throughout the cardiac cycle (P=0.290). Specifically, at no measured point at any level of cardiac filling volume was the pulmonary capillary wedge pressure of the seniors lower than that of the young subjects. CONCLUSIONS: We conclude that the age-related echocardiographic change of decreasing E/A ratio is not the result of a lowering of early diastolic left atrial pressure. PMID- 15781733 TI - In vivo characterization of coronary atherosclerotic plaque by use of optical coherence tomography. AB - BACKGROUND: The current understanding of the pathophysiology of coronary artery disease is based largely on postmortem studies. Optical coherence tomography (OCT) is a high-resolution ( approximately 10 microm), catheter-based imaging modality capable of investigating detailed coronary plaque morphology in vivo. METHODS AND RESULTS: Patients undergoing cardiac catheterization were enrolled and categorized according to their clinical presentation: recent acute myocardial infarction (AMI), acute coronary syndromes (ACS) constituting non-ST-segment elevation AMI and unstable angina, or stable angina pectoris (SAP). OCT imaging was performed with a 3.2F catheter. Two observers independently analyzed the images using the previously validated criteria for plaque characterization. Of 69 patients enrolled, 57 patients (20 with AMI, 20 with ACS, and 17 with SAP) had analyzable images. In the AMI, ACS, and SAP groups, lipid-rich plaque (defined by lipid occupying > or =2 quadrants of the cross-sectional area) was observed in 90%, 75%, and 59%, respectively (P=0.09). The median value of the minimum thickness of the fibrous cap was 47.0, 53.8, and 102.6 microm, respectively (P=0.034). The frequency of thin-cap fibroatheroma (defined by lipid-rich plaque with cap thickness < or =65 microm) was 72% in the AMI group, 50% in the ACS group, and 20% in the SAP group (P=0.012). No procedure-related complications occurred. CONCLUSIONS: OCT is a safe and effective modality for characterizing coronary atherosclerotic plaques in vivo. Thin-cap fibroatheroma was more frequently observed in patients with AMI or ACS than SAP. This is the first study to compare detailed in vivo plaque morphology in patients with different clinical presentations. PMID- 15781735 TI - Reconstituted high-density lipoproteins inhibit the acute pro-oxidant and proinflammatory vascular changes induced by a periarterial collar in normocholesterolemic rabbits. AB - BACKGROUND: HDLs have antiinflammatory and antioxidant properties in vitro. This study investigates these properties in vivo. METHODS AND RESULTS: Chow-fed, normocholesterolemic New Zealand White rabbits received a daily infusion of (1) saline, (2) reconstituted HDL (rHDL) containing 25 mg apolipoprotein (apo) A-I and 50 mg of either 1-palmitoyl-2-linoleoyl phosphatidylcholine (PLPC) or 1,2 dipalmitoyl phosphatidylcholine (DPPC), (3) 25 mg lipid-free apoA-I, or (4) 50 mg of either PLPC-small unilamellar vesicles (SUVs) or DPPC-SUVs on each of 3 consecutive days. Nonocclusive carotid periarterial collars were implanted after the second dose of treatment. Forty-eight hours after insertion of the collars, the arteries were removed and analyzed for the presence of reactive oxygen species, the infiltration of neutrophils, and the expression of adhesion proteins and chemokines. Insertion of the periarterial collar induced a 4.1-fold increase in presence of vascular wall reactive oxygen species. This effect was completely abolished in the animals infused with rHDL. The periarterial collar was associated with a dense infiltration of the arterial wall by polymorphonuclear leukocytes. This infiltration was inhibited by 73% to 94% in the animals infused with rHDL, by 75% in the animals infused with lipid-free apoA-I, and by 51% to 65% in animals infused with SUVs. There were no significant differences between the effects of PLPC and DPPC in either the rHDL or SUVs. Endothelial expression of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, and monocyte chemoattractant protein-1 was also increased by the collar insertion and inhibited by rHDL, lipid-free apoA-I, and, to a lesser extent, also by the SUVs. CONCLUSIONS: Infusion of rHDL, apoA-I, and phospholipid-SUVs inhibits the early pro-oxidant and proinflammatory changes induced by a periarterial collar in normocholesterolemic rabbits. PMID- 15781736 TI - Risk of worsening renal function with nesiritide in patients with acutely decompensated heart failure. AB - BACKGROUND: Renal function is an important prognostic factor for patients with acutely decompensated heart failure (ADHF). We investigated the renal effects of nesiritide as treatment for ADHF. METHODS AND RESULTS: Randomized clinical trials comparing nesiritide with either placebo or active control for ADHF were identified by electronic and manual searches and thorough review of US Food and Drug Administration files available via the website. Worsening renal function was defined as an increase in serum creatinine >0.5 mg/dL. Relative risk across all studies was determined by meta-analysis with Mantel-Haenszel fixed-effects models (RR(MH)). Risk of dialysis and medical intervention for worsening renal function were compared between therapies. Frequency of worsening renal function was determined from 5 randomized studies that included 1269 patients. Use of Food and Drug Administration-approved doses of nesiritide (< or =0.03 microg x kg(-1) x min(-1)) significantly increased the risk of worsening renal function compared with non-inotrope-based control (RR(MH), 1.52; 95% CI, 1.16 to 2.00; P=0.003) or any control therapy, including non-inotrope- and inotrope-based therapies (RR(MH), 1.54; 95% CI, 1.19 to 1.98; P=0.001). Even low-dose nesiritide (< or =0.015 microg x kg(-1) x min(-1)) significantly increased risk (P=0.012 and P=0.006 compared with non-inotrope- and inotrope-based controls, respectively), as did nesiritide administered at any dose up to 0.06 microg x kg(-1) x min(-1) (P=0.002 and P=0.001, respectively). There was no difference in the need for dialysis between therapies. CONCLUSIONS: Nesiritide significantly increases the risk of worsening renal function in patients with ADHF. Whether worsening renal function reflects hemodynamic effect or renal injury is unknown, but the prognostic importance of worsening renal function suggests the need for further investigation in appropriately powered clinical trials. PMID- 15781737 TI - Adenovirus-mediated overexpression of diacylglycerol kinase-zeta inhibits endothelin-1-induced cardiomyocyte hypertrophy. AB - BACKGROUND: Diacylglycerol (DAG) is a lipid second messenger that transiently accumulates in cells stimulated by endothelin-1 (ET-1) and other Galphaq protein coupled receptor agonists. Diacylglycerol kinase (DGK) is thought to be an enzyme that controls the cellular levels of DAG by converting it to phosphatidic acid; however, the functional role of DGK has not been examined in cardiomyocytes. Because DGK inactivates DAG, a strong activator of protein kinase C (PKC), we hypothesized that DGK inhibited ET-1-induced activation of a DAG-PKC signaling cascade and subsequent cardiomyocyte hypertrophy. METHODS AND RESULTS: Real-time reverse transcription-polymerase chain reaction demonstrated a significant increase of DGK-zeta mRNA by ET-1 in cardiomyocytes. To determine the functional role of DGK-zeta, we overexpressed DGK-zeta in cardiomyocytes using a recombinant adenovirus encoding rat DGK-zeta (Ad-DGKzeta). ET-1-induced translocation of PKC epsilon was blocked by Ad-DGKzeta (P<0.01). Ad-DGKzeta also inhibited ET-1 induced activation of extracellular signal-regulated kinase (P<0.01). Luciferase reporter assay revealed that ET-1-mediated increase of activator protein-1 (AP1) DNA-binding activity was significantly inhibited by DGK-zeta (P<0.01). In cardiomyocytes transfected with DGK-zeta, ET-1 failed to cause gene induction of atrial natriuretic factor, increases in [3H]-leucine uptake, and increases in cardiomyocyte surface area. CONCLUSIONS: We demonstrated for the first time that DGK-zeta blocked ET-1-induced activation of the PKC-epsilon-ERK-AP1 signaling pathway, atrial natriuretic factor gene induction, and resultant cardiomyocyte hypertrophy. DGK-zeta might act as a negative regulator of hypertrophic program in response to ET-1, possibly by controlling cellular DAG levels. PMID- 15781738 TI - Sirolimus-eluting stents to abolish intimal hyperplasia and improve flow in porcine arteriovenous grafts: a 4-week follow-up study. AB - BACKGROUND: The patency of arteriovenous (AV) expanded polytetrafluoroethylene (ePTFE) hemodialysis grafts is severely compromised by intimal hyperplasia (IH) at the venous anastomosis and in the venous outflow tract. We addressed the potential of primary placement of a sirolimus-eluting stent (SES) in a validated porcine model. METHODS AND RESULTS: In 25 pigs, ePTFE AV grafts were created bilaterally between the carotid artery and the jugular vein, whereupon a self expandable nitinol stent (14 SESs and 11 bare-metal stents) was implanted over the venous anastomosis in 1 of the 2 grafts. After exclusion of technical failures and 1 unilateral occlusion, 16 pigs (9 SESs and 7 bare-metal stents) were included for further analysis. After 28 days, we measured graft flow and performed quantitative angiography. The pigs were then euthanized, and grafts with adjacent vessels were excised for histological analysis. Minimal luminal diameter was substantially larger in the SES group compared with unstented controls (5.9+/-0.2 versus 3.8+/-0.4 mm, respectively, P=0.01), which was accompanied by more prominent graft flow (SES, 1360+/-89 mL/min versus unstented, 861+/-83 mL/min, P=0.05). IH at the venous anastomosis was 77% less in the SES group compared with unstented controls (0.44+/-0.05 versus 1.92+/-0.5 mm2, respectively, P=0.01), whereas IH increased markedly when bare-metal stents were used (5.7+/-1.4 mm2, P=0.05). CONCLUSIONS: SESs in the venous outflow of AV grafts significantly reduce IH and increase vessel diameter and graft flow compared with unstented grafts. These findings suggest that SESs have the potential to improve primary patency of AV grafts in hemodialysis patients. PMID- 15781739 TI - Increased mortality and aggravation of heart failure in estrogen receptor-beta knockout mice after myocardial infarction. AB - BACKGROUND: Lower mortality rates among women with chronic heart failure than among men may depend in part on the action of female sex hormones, especially estrogens. The biological effects of estrogens are mediated by 2 distinct estrogen receptor (ER) subtypes (ERalpha and ERbeta). The present study was undertaken to determine the role of ERbeta in the development of chronic heart failure after experimental myocardial infarction (MI). METHODS AND RESULTS: Female ERbeta null mice (BERKO(Chapel Hill)) and wild-type littermates (WT) were ovariectomized, given 17beta-estradiol, and subjected to chronic anterior MI (MI; BERKO n=31, WT n=30) or sham operation (sham; BERKO n=14, WT n=14). At 8 weeks after MI, both genotypes revealed left ventricular remodeling and impaired contractile function at similar average infarct size (BERKO-MI 32.9+/-5% versus WT-MI 33.0+/-4%); however, BERKO mice showed increased mortality (BERKO-MI 42% versus WT-MI 23%), increased body weight and fluid retention (P<0.01), higher ventricular pro-ANP expression (BERKO-MI 27.9-fold versus sham, WT-MI 5.2-fold versus sham; BERKO-MI versus WT-MI P<0.001), higher atrial natriuretic peptide serum levels, and increased phospholamban expression (P<0.05) compared with WT mice. CONCLUSIONS: Systemic deletion of ERbeta in female mice increases mortality, aggravates clinical and biochemical markers of heart failure, and contributes to impaired expression of Ca(2+)-handling proteins in chronic heart failure after MI. Further studies are required to delineate the relative importance of cardiac and vascular effects of ERbeta and the role of ERalpha in the development of heart failure. PMID- 15781740 TI - Limited exercise capacity in heterozygous manganese superoxide dismutase gene knockout mice: roles of superoxide anion and nitric oxide. AB - BACKGROUND: We have reported that there is a limitation of exercise capacity in mice with defects in the expression of endothelial nitric oxide (NO) synthase, which is associated with a greater increase in whole-body oxygen consumption (VO2). We hypothesized that in states in which superoxide anion (O2-) is increased, especially in the mitochondria, whole-body VO2 will be increased because of the inactivation of NO, and consequently, exercise capacity will be reduced. METHODS AND RESULTS: Heterozygous manganese superoxide anion dismutase (SOD2) gene-knockout mice (SOD2+/-), in which SOD2 activity is reduced by 30% to 80%, and wild-type control mice (SOD2+/+) were treadmill-tested to measure indices defining exercise capacity. Tempol was given to each mouse for 7 days by an intraperitoneal injection to scavenge O2- before a second treadmill testing. VO2 and carbon dioxide production (VCO2) at rest were increased in SOD2+/-. The work (vertical distance run x body weight) to exhaustion was decreased in SOD2+/ . When the maximum VO2 and VCO2 were corrected to per work unit, they were increased in SOD2+/-. Tempol normalized basal VO2 and VCO2 and improved the work to exhaustion and corrected VO2 and VCO2 in SOD2+/-. VO2 of skeletal muscle was measured in vitro. Bradykinin-induced reduction in VO2 in vitro was attenuated in SOD2+/-, and was acutely restored by Tempol. There was a decrease in SOD2 protein level and a concomitant increase in lucigenin-detectable O2- production in skeletal muscle from SOD2+/-. CONCLUSIONS: These results suggest that exercise capacity is reduced in conditions in which superoxide anion is increased, and this is associated with a greater increase in whole-body oxygen consumption in SOD2+/- compared with SOD2+/+. PMID- 15781741 TI - Effects of intravenous levosimendan on human coronary vasomotor regulation, left ventricular wall stress, and myocardial oxygen uptake. AB - BACKGROUND: Levosimendan is a calcium-sensitizing agent and an inodilator under current investigation in the treatment of decompensated heart failure. The effects of intravenous levosimendan on the human coronary vasculature, together with myocardial wall stress and oxygen uptake, have not been adequately studied. METHODS AND RESULTS: Ten adult patients underwent right- and left-heart catheterization. Baseline coronary blood flow was determined with quantitative coronary angiography and an intracoronary Doppler-tipped guidewire. Myocardial oxygen uptake was measured with a coronary sinus catheter. Echocardiography was performed before and 30 minutes after an intravenous infusion of levosimendan (24 microg/kg bolus over 10 minutes) was begun. Pulmonary capillary wedge decreased 37% (P=0.009), cardiac output increased 9% (P=0.04), and systemic vascular resistance decreased 18% (P<0.001). Left ventricular ejection fraction increased 20% (P=0.009), and meridional systolic wall stress decreased 48% (P=0.009). Coronary artery diameter increased 10% at 15 minutes (P=0.001) and 11% at 30 minutes (P=0.01). Coronary artery velocity increased 10% over baseline (P=0.04). Coronary blood flow increased 45% (P=0.02), whereas coronary resistance decreased 36% at 30 minutes (P=0.03). Myocardial oxygen extraction decreased 9% at 30 minutes (P=0.04). CONCLUSIONS: Levosimendan given intravenously exerts vasodilator effects on human coronary conductance and resistance arteries. Despite a decrease in coronary perfusion pressure, coronary blood flow is increased. A reduction in coronary vascular resistance and a decrease in coronary venous oxygen content indicate primary coronary vasodilation by levosimendan. Improved left ventricular systolic function and decreased myocardial oxygen extraction suggest improved myocardial efficiency. PMID- 15781742 TI - Risk of cardiovascular disease by hysterectomy status, with and without oophorectomy: the Women's Health Initiative Observational Study. AB - BACKGROUND: Cardiovascular disease (CVD) is a leading cause of morbidity and mortality in women and may vary by hysterectomy (or oophorectomy) status. This study compared CVD risk factors and rates between postmenopausal women who had and had not undergone hysterectomy, with or without oophorectomy. METHODS AND RESULTS: This analysis was conducted on 89 914 women in the Women's Health Initiative (WHI) Observational Study. Participants reported demographic characteristics, medical history, dietary habits, physical activity, medications, and previous hysterectomy (with or without oophorectomy). Baseline weight, height, waist circumference, and blood pressure were measured. CVD events were ascertained during 5.1 years of mean follow-up and adjudicated with standard criteria. Black, Hispanic, and American Indian women had higher rates of hysterectomy than white women (52.9%, 44.6%, and 49.2% versus 40.0%, respectively), and Asian/Pacific Islander women had lower rates (33.8%). Women with a hysterectomy (regardless of oophorectomy status) had an adverse risk profile at baseline compared with women with no hysterectomy, including a higher proportion of hypertension, diabetes, high cholesterol, obesity, and lower education, income, and physical activity (all P<0.01). Total mortality and fatal and nonfatal CVD were higher among women with a hysterectomy. Hysterectomy (regardless of oophorectomy status) was a significant predictor of CVD (HR: 1.26, P<0.001). After adjustment for demographic variables and CVD risk factors, the effect was reduced and nonsignificant. CONCLUSIONS: Women with a hysterectomy had a worse risk profile and higher prevalence and incidence of CVD in this cohort. Multivariate models suggest that hysterectomy is not the major determinant of this outcome; rather, CVD risk may be due to the more adverse initial risk profile of women who had undergone hysterectomy. PMID- 15781743 TI - Lipoprotein(a) and apolipoprotein(a) isoforms: no association with coronary artery calcification in the Dallas Heart Study. AB - BACKGROUND: Elevated plasma levels of lipoprotein(a) [Lp(a)] are an independent risk factor for cardiovascular disease in whites. Blacks have 2- to 3-fold higher plasma levels of Lp(a) than whites and yet do not have a correspondingly higher rate of coronary events. It remains unclear whether elevated plasma levels of Lp(a) are an independent risk factor for coronary atherosclerosis in individuals of African descent. METHODS AND RESULTS: The relationship between plasma levels of Lp(a), apolipoprotein(a) isoform sizes, and the presence of coronary calcium was examined in 761 blacks and 527 whites (men aged >40 years, women aged >45 years) from a population-based sample. No relationship was found between plasma levels of Lp(a), apolipoprotein(a) isoform size, or a combination of these 2 variables and coronary artery calcium (CAC) in whites or blacks. No correlation was observed between plasma levels of Lp(a) and coronary calcium scores in any group, although all black men with very high plasma levels of Lp(a) (>300 micromol/L; n=7) were CAC-positive. Whites with high plasma levels of Lp(a) plus elevated plasma levels of LDL cholesterol (men) or reduced levels of HDL cholesterol (men and women) or who smoked (women) had a higher prevalence of CAC. In contrast, no joint effects between plasma levels of Lp(a) and other cardiovascular risk factors on coronary calcium were found in blacks. CONCLUSIONS: No consistent independent relationship between plasma levels of Lp(a) or apolipoprotein(a) isoform size and coronary calcium was found in whites or blacks. PMID- 15781744 TI - Renin-aldosterone paradox and perturbed blood volume regulation underlying postural tachycardia syndrome. AB - BACKGROUND: Patients with postural tachycardia syndrome (POTS) experience considerable disability, but in most, the pathophysiology remains obscure. Plasma volume disturbances have been implicated in some patients. We prospectively tested the hypothesis that patients with POTS are hypovolemic compared with healthy controls and explored the role of plasma renin activity and aldosterone in the regulation of plasma volume. METHODS AND RESULTS: Patients with POTS (n=15) and healthy controls (n=14) underwent investigation. Heart rate (HR), blood pressure (BP), plasma renin activity, and aldosterone were measured with patients both supine and upright. Blood volumes were measured with 131I-labeled albumin and hematocrit. Patients with POTS had a higher orthostatic increase in HR than controls (51+/-18 versus 16+/-10 bpm, P<0.001). Patients with POTS had a greater deficit in plasma volume (334+/-187 versus 10+/-250 mL, P<0.001), red blood cell volume (356+/-128 versus 218+/-140 mL, P=0.010), and total blood volume (689+/-270 versus 228+/-353 mL, P<0.001) than controls. Despite the lower plasma volume in patients with POTS, there was not a compensatory increase in plasma renin activity (0.79+/-0.58 versus 0.79+/-0.74 ng x mL(-1) x h(-1), P=0.996). There was a paradoxically low level of aldosterone in the patients with POTS (190+/-140 pmol/L versus 380+/-230 pmol/L; P=0.017). CONCLUSIONS: Patients with POTS have paradoxically unchanged plasma renin activity and low aldosterone given their marked reduction in plasma volume. These patients also have a significant red blood cell volume deficit, which is regulated by the renal hormone erythropoietin. These abnormalities suggest that the kidney may play a key role in the pathophysiology of POTS. PMID- 15781745 TI - Treatment of aortic disease in patients with Marfan syndrome. PMID- 15781746 TI - Images in cardiovascular medicine. Follow-up of cardiac sarcoidosis by magnetic resonance imaging. PMID- 15781747 TI - Images in cardiovascular medicine. Himalayan T waves in the congenital long-QT syndrome. PMID- 15781748 TI - Polymerase chain reaction to diagnose infective endocarditis: will it replace blood cultures? PMID- 15781749 TI - Short- and long-term clinical outcome after drug-eluting stent implantation for the percutaneous treatment of left main coronary artery disease: insights from the Rapamycin-Eluting and Taxus Stent Evaluated At Rotterdam Cardiology Hospital registries (RESEARCH and T-SEARCH). AB - BACKGROUND: The impact of drug-eluting stent (DES) implantation on the incidence of major adverse cardiovascular events in patients undergoing percutaneous intervention for left main (LM) coronary disease is largely unknown. METHODS AND RESULTS: From April 2001 to December 2003, 181 patients underwent percutaneous coronary intervention for LM stenosis at our institution. The first cohort consisted of 86 patients (19 protected LM) treated with bare metal stents (pre DES group); the second cohort comprised 95 patients (15 protected LM) treated exclusively with DES. The 2 cohorts were well balanced for all baseline characteristics. At a median follow-up of 503 days (range, 331 to 873 days), the cumulative incidence of major adverse cardiovascular events was lower in the DES cohort than in patients in the pre-DES group (24% versus 45%, respectively; hazard ratio [HR], 0.52 [95% CI, 0.31 to 0.88]; P=0.01). Total mortality did not differ between cohorts; however, there were significantly lower rates of both myocardial infarction (4% versus 12%, respectively; HR, 0.22 [95% CI, 0.07 to 0.65]; P=0.006) and target vessel revascularization (6% versus 23%, respectively; HR, 0.26 [95% CI, 0.10 to 0.65]; P=0.004) in the DES group. On multivariate analysis, use of DES, Parsonnet classification, troponin elevation at entry, distal LM location, and reference vessel diameter were independent predictors of major adverse cardiovascular events. CONCLUSIONS: When percutaneous coronary intervention is undertaken at LM lesions, routine DES implantation, which reduces the cumulative incidence of myocardial infarction and the need for target vessel revascularization compared with bare metal stents, should currently be the preferred strategy. PMID- 15781750 TI - Randomized, blinded trial comparing fondaparinux with unfractionated heparin in patients undergoing contemporary percutaneous coronary intervention: Arixtra Study in Percutaneous Coronary Intervention: a Randomized Evaluation (ASPIRE) Pilot Trial. AB - BACKGROUND: Factor Xa plays a central role in the generation of thrombin, making it a novel target for treatment of arterial thrombosis. Fondaparinux is a synthetic factor Xa inhibitor that has been shown to be superior to standard therapies for the prevention of venous thrombosis. We performed a randomized trial to determine the safety and feasibility of fondaparinux in the percutaneous coronary intervention (PCI) setting. METHODS AND RESULTS: A total of 350 patients undergoing elective or urgent PCI were randomized in a blinded manner to receive unfractionated heparin (UFH), 2.5 mg fondaparinux IV, or 5.0 mg fondaparinux IV. Randomization was stratified for planned or no planned use of glycoprotein (GP) IIb/IIIa antagonists. The primary safety outcome was total bleeding, which was a combination of major and minor bleeding events. The incidence of total bleeding was 7.7% in the UFH group and 6.4% in the combined fondaparinux groups (hazard ratio, 0.81; 95% confidence interval, 0.35 to 1.84; P=0.61). Bleeding was less common in the 2.5-mg fondaparinux group compared with the 5-mg fondaparinux group (3.4% versus 9.6%, P=0.06). The composite efficacy outcome of all-cause mortality, myocardial infarction, urgent revascularization, or need for a bailout GPIIb/IIIa antagonist was 6.0% in the UFH group and 6.0% in the fondaparinux group, with no significant difference in efficacy among the fondaparinux doses compared with UFH. Coagulation marker analysis at 6 and 12 hours after PCI demonstrated that fondaparinux was superior to UFH in inducing a sustained reduction in markers of thrombin generation, as measured by prothrombin fragment F1.2 (P=0.02). CONCLUSIONS: In this pilot study of patients undergoing contemporary PCI, factor Xa inhibition with the synthetic anticoagulant fondaparinux in doses of 2.5 and 5.0 mg was comparable to UFH for clinical safety and efficacy outcomes. These data form the basis for further evaluation of fondaparinux in arterial thrombosis. PMID- 15781751 TI - Blockade of endogenous cytokines mitigates neointimal formation in obese Zucker rats. AB - BACKGROUND: It is well known that diabetes mellitus is a major risk factor for vascular diseases such as atherosclerosis and restenosis after angioplasty. It has become clear that advanced glycation end products (AGE) and their receptor (RAGE) are implicated in vascular diseases, especially in diabetes mellitus. Nevertheless, the mechanisms by which diabetes mellitus is often associated with vascular diseases remain unclear. METHODS AND RESULTS: To study the role of endogenous cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 in the development of vascular diseases and in the expression of RAGE, we used semapimod, a pharmacological inhibitor of cytokine production, and examined its effect on neointimal formation in the femoral artery of obese Zucker (OZ) rats. We also used an adenovirus construct expressing a dominant negative mutant of the receptor for TNF-alpha (AdTNFRDeltaC) to block the action of endogenous TNF-alpha. Semapimod significantly suppressed neointimal formation and RAGE expression in OZ rats compared with untreated OZ rats. This inhibitory effect of semapimod on neointimal formation was overcome by infection of an adenovirus expressing RAGE into the femoral artery of OZ rats. Furthermore, AdTNFRDeltaC infection significantly suppressed neointimal formation and RAGE expression in the femoral artery of OZ rats. CONCLUSIONS: These results suggest that endogenous cytokines, especially TNF-alpha, were implicated in neointimal formation in OZ rats and that RAGE was a mediator of the effect of these cytokines on neointimal formation. PMID- 15781752 TI - Amelioration of pulmonary emphysema by in vivo gene transfection with hepatocyte growth factor in rats. AB - BACKGROUND: Hepatocyte growth factor (HGF) is an important mitogen and morphogen that contributes to the repair process after lung injury. The goal of the present study was to characterize its role in pulmonary emphysema, which may lead to the development of new treatment strategies with HGF. METHODS AND RESULTS: HGF mRNA and protein levels in lung tissue and plasma from elastase-induced emphysema rats transiently increased, then declined significantly to below the basal level in a time-dependent manner (P<0.01). Furthermore, changes in HGF were correlated with histologically progressive emphysematous changes and deterioration in pulmonary physiology. Use of the HVJ (hemagglutinating virus of Japan) envelope method resulted in successful transfection of cDNA encoding human HGF, as demonstrated by an efficient expression of HGF in alveolar endothelial and epithelial cells. Transfection of HGF resulted in a more extensive pulmonary vasculature and inhibition of alveolar wall cell apoptosis, and those effects led to improved exercise tolerance and gas exchange (P<0.05), which persisted for more than 1 month. CONCLUSIONS: Decreased HGF expression due to a failure in sustained endogenous production after injury was associated with emphysema-related histopathologic and physiological changes in the present rat model. In addition, induction of HGF expression by a gene-transfection method resulted in improved pulmonary function via inhibition of alveolar cell apoptosis, enhancement of alveolar regeneration, and promotion of angiogenesis. PMID- 15781753 TI - Plaque rupture after short periods of fat feeding in the apolipoprotein E knockout mouse: model characterization and effects of pravastatin treatment. AB - BACKGROUND: These studies examined the early time course of plaque development and destabilization in the brachiocephalic artery of the apolipoprotein E knockout mouse, the effects of pravastatin thereon, and the effects of pravastatin on established unstable plaques. METHODS AND RESULTS: Male apolipoprotein E-knockout mice were fed a high-fat, cholesterol-enriched diet from the age of 8 weeks. Animals were euthanized at 1-week intervals between 4 and 9 weeks of fat feeding. Acutely ruptured plaques were observed in the brachiocephalic arteries of 3% of animals up to and including 7 weeks of fat feeding but in 62% of animals after 8 weeks, which suggests that there is a sharp increase in the number of plaque ruptures at 8 weeks. These acute plaque ruptures then appear to heal and form buried fibrous caps; after 9 weeks of fat feeding, mice had 1.05+/-0.15 buried fibrous caps at a single site in the brachiocephalic artery. Pravastatin (40 mg/kg of body weight per day for 9 weeks; resultant plasma concentration 16+/-4 nmol/L) had no effect on plasma cholesterol concentration in fat-fed apolipoprotein E-knockout mice but reduced the number of buried fibrous caps by 43% (P<0.0001). In longer-term experiments, the delay of pravastatin treatment until unstable plaques had developed reduced the incidence of acute plaque rupture by 36% (P<0.0001). CONCLUSIONS: Plaque rupture occurs at high frequency in the brachiocephalic arteries of male apolipoprotein E-knockout mice after 8 weeks of fat feeding. Pravastatin treatment inhibits early plaque rupture and is also effective when begun after unstable plaques have developed. PMID- 15781754 TI - Overexpression of dimethylarginine dimethylaminohydrolase reduces tissue asymmetric dimethylarginine levels and enhances angiogenesis. AB - BACKGROUND: This study was designed to determine whether overexpression of the enzyme dimethylarginine dimethylaminohydrolase (DDAH) could enhance angiogenesis by reducing levels of the endogenous nitric oxide synthase (NOS) inhibitor asymmetric dimethylarginine (ADMA). METHODS AND RESULTS: In DDAH1 transgenic (TG) and wild-type mice (each n=42), the role of DDAH overexpression on angiogenesis was studied by use of the disk angiogenesis system and a murine model of hindlimb ischemia (each n=21). After surgery, animals were treated with either PBS or the NOS inhibitors ADMA or N(omega)-nitro-L-arginine methyl ester (L-NAME; each 250 micromol x kg(-1) x d(-1)) by use of osmotic minipumps (each n=7). L-NAME was chosen to study an inhibitor that is not degraded by DDAH. Neovascularization in the disk angiogenesis system was impaired by both NOS inhibitors; however, TG animals were resistant to the effects of ADMA on neovascularization. Similarly, TG mice were more resistant to the inhibitory effect of ADMA on angioadaptation (angiogenesis and arteriogenesis) after hindlimb ischemia, as assessed by fluorescent microsphere studies and postmortem microangiograms. Enhanced neovascularization and limb perfusion in TG mice were associated with reduced plasma and tissue ADMA levels and enhanced tissue NOS enzyme activity. CONCLUSIONS: We describe a novel mechanism by which DDAH regulates postnatal neovascularization. Therapeutic manipulation of DDAH expression or activity may represent a novel approach to restore tissue perfusion. PMID- 15781755 TI - HMG-CoA reductase inhibition reduces monocyte CC chemokine receptor 2 expression and monocyte chemoattractant protein-1-mediated monocyte recruitment in vivo. AB - BACKGROUND: The migration of circulating monocytes to the arterial wall during atherogenesis is largely modulated by activation of the CC chemokine receptor 2 (CCR2), a dominant monocyte chemotaxis receptor. The present study investigated whether 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibition affects CCR2 gene expression and CCR2-dependent monocyte recruitment. METHODS AND RESULTS: Competitive reverse transcription-polymerase chain reaction analysis and flow cytometry showed that simvastatin, an HMG-CoA reductase inhibitor, dose dependently reduced monocyte CCR2 mRNA and protein expression. Treatment of 21 normocholesterolemic men with simvastatin (20 mg/d for 2 weeks) decreased CCR2 protein and mRNA expression in circulating monocytes. Promoter and electrophoretic mobility shift assays showed that simvastatin activated a peroxisome proliferator response element in THP-1 monocytes. Moreover, simvastatin-induced CCR2 downregulation was completely reversed by the synthetic peroxisome proliferator-activated receptor-gamma antagonist GW9662. Simvastatin treated monocytes showed little chemotaxis movement in response to monocyte chemoattractant protein-1 (MCP-1), a specific CCR2 ligand. Treatment of C57/BL6 mice with simvastatin (0.2 microg/g body weight IP, daily for 1 week) inhibited transmigration of CD80+ monocytes to the MCP-1-injected intraperitoneal space. Moreover, few circulating inflammatory cells from simvastatin-treated Sprague Dawley rats (0.2 microg/g body weight IP, daily for 2 weeks) were recruited to the aortic wall of hypercholesterolemic littermates. CONCLUSIONS: The inhibition of CCR2/MCP-1-dependent monocyte recruitment by simvastatin may prevent excessive accumulation of monocytes in the arterial wall during atherogenesis. PMID- 15781756 TI - Metabolic syndrome: a comprehensive perspective based on interactions between obesity, diabetes, and inflammation. PMID- 15781758 TI - Chemoprevention of head and neck cancer with retinoids: a negative result. AB - OBJECTIVE: To determine whether isotretinoin (or 13-cis-retinoic acid) decreases the risk of second primary cancers in patients previously treated for cure of head and neck squamous cell carcinoma. DESIGN: Randomized, double-blind, placebo controlled trial. SETTING: Two head and neck multidisciplinary cancer clinics in university teaching hospitals taking cases from 4 to 5 million people in Queensland, Australia, combined to enter appropriate patients into this trial. PATIENTS: One hundred fifty-one patients with their first head and neck squamous cell carcinoma treated with high expectation for cure and living close by. They were randomized into 3 arms to receive 3 years of treatment. INTERVENTIONS: Patients took isotretinoin at a high dose (1.0 mg/kg per day) or a moderate dose (0.5 mg/kg per day) or placebo. Group 1 took the high dose for 1 year and then the moderate dose for 2 years. Group 2 took the moderate dose for 3 years. Group 3 took placebo for 3 years. MAIN OUTCOME MEASURES: The diagnosis of a second primary malignancy of the head and neck, lung, or bladder was regarded as the end point signifying failure of therapy. Issues of drug adverse effect profile and impact on survival were measured. RESULTS: There was no significant difference in the occurrence of second primary disease (P = .90), the recurrence of primary disease (P = .70), or disease-free time (P = .80) between the treatment and nontreatment arms. Numbers were too small to find differences in survival. CONCLUSION: With evidence that retinoid treatment adversely affects survival of lung cancer and with this drug not significantly decreasing the incidence of second primary tumors of head and neck squamous cell carcinoma, the use of this drug in head and neck cancer patients for second cancer prophylaxis is not indicated. PMID- 15781759 TI - The effect of tetrathiomolybdate on cytokine expression, angiogenesis, and tumor growth in squamous cell carcinoma of the head and neck. AB - OBJECTIVE: To assess the effect of tetrathiomolybdate on cytokine expression, angiogenesis, and tumor growth rate in human squamous cell carcinoma (SCC). DESIGN: Three human SCC cell lines were used in this study for both in vitro and in vivo investigations. Conditioned media from untreated and tetrathiomolybdate treated cell lines were compared with regard to cytokine levels, endothelial cell chemotaxis, endothelial cell tubule formation, and migration and the ability to induce angiogenesis in a rat aortic ring array. In vivo UM-SCC-38 was seeded onto tissue-engineered scaffolds and surgically implanted into the flanks of immunodeficient mice. Tumor growth rates and the level of angiogenesis were compared after 2 weeks of therapy. SETTING: A tertiary care facility. RESULTS: In this study, we demonstrate that tetrathiomolybdate significantly decreases the secretion of interleukin 6 and basic fibroblast growth factor by head and neck SCC (HNSCC) cell lines in vitro. Furthermore, we demonstrate that tetrathiomolybdate significantly decreases the secretion of interleukin 6 and basic fibroblast growth factor by HNSCC cell lines in vitro. Furthermore, tetrathiomolybdate treatment of HNSCC cell lines results in significantly decreased endothelial cell chemotaxis, tubule formation, and neovascularization in a rat aortic ring assay. This in vitro evidence of decreased angiogenesis by tetrathiomolybdate is confirmed in vivo by using a severe combined immunodeficiency disorder mouse model in which tetrathiomolybdate therapy is shown to prevent human blood vessel formation. Finally, human HNSCC implanted into immunodeficient mice grow to a much larger size in untreated mice compared with those treated with 0.7 mL/kg per day of oral tetrathiomolybdate. CONCLUSIONS: These findings illustrate the ability of tetrathiomolybdate to down regulate proinflammatory and proangiogenic cytokines in HNSCC. These observations are potentially exciting from a clinical perspective because a global decrease in these cytokines may decrease tumor aggressiveness and reverse the resistance to chemotherapy and radiation therapy seen in this tumor type. PMID- 15781760 TI - Curative surgery for recurrent nasopharyngeal carcinoma via the infratemporal fossa approach. AB - OBJECTIVE: To determine the proper indications (and thus define good candidates) for the infratemporal fossa approach (ITFA) in the treatment of recurrent nasopharyngeal carcinoma (NPC). DESIGN: Retrospective analysis of 11 consecutive patients who had recurrent NPC after radiation therapy failure and were treated with curative surgery via the ITFA approach from July 1, 1993, to November 20, 1999. The mean follow-up was 32.5 months (range, 9-56 months). SETTING: Academic tertiary referral center. RESULTS: Patients who had a recurrent tumor confined to the Rosenmuller fossa (rT1; n = 3) or extending to the parapharynx (rT2; n = 3) maintained a clear surgical margin and were disease free. However, patients who had rT1 (n = 1) or rT2 (n = 1) tumors crossing the midline of the posterior nasopharyngeal wall had surgical margin involvement and their tumors recurred, and all patients with rT3 (n = 2) or rT4 (n = 1) tumors eventually died of the disease. CONCLUSIONS: Resection of recurrent NPC via ITFA is useful for tumors located in the Rosenmuller fossa with or without parapharyngeal extension. However, tumors extending to the contralateral nasopharyngeal mucosa and tumors at an advanced stage are not suitable indications for ITFA. PMID- 15781761 TI - Assessment of construct validity of the endoscopic sinus surgery simulator. AB - OBJECTIVE: To study the relationship between performance on an endoscopic sinus surgery simulator (ES3) and fundamental perceptual, visuospatial, and psychomotor abilities. DESIGN: Validation study. SETTING: Tertiary care medical center. PARTICIPANTS: Thirty-four medical students and 4 otolaryngology residents voluntarily enrolled. INTERVENTIONS: Subjects performed tasks on the ES3, minimally invasive surgical trainer virtual reality (MIST-VR), pictorial surface orientation (PicSOr), and 3 visuospatial tests (cube comparison, card rotation, and map planning). MAIN OUTCOME MEASURES: The MIST-VR was scored for time, task error, economy of hand movement, economy of diathermy, and total score. Scores were generated for the PicSOr task and visuospatial tests. Scores were correlated with time, accuracy, and total subscore on navigation, injection, and dissection tasks, as well as hazard score and total trial score on the ES3. RESULTS: The PicSOr score was statistically significantly correlated with the hazard score on the ES3 (r = 0.50, P < .001). Cube comparison (r = 0.43, P < .01) and card rotation (r = 0.45, P < .01) scores correlated significantly with the ES3 trial score, as did the MIST-VR total score and the ES3 trial score (r = 0.57, P < .001). In a multiple regression model, the PicSOr, cube comparison, and MIST-VR total scores were statistically significant predictors of ES3 performance (r = 0.63, P < .01). CONCLUSIONS: Scores on the ES3 correlate strongly with scores on previously validated measures of perceptual, visuospatial, and psychomotor performance. The ES3 provides a reliable assessment of factors that are important to the acquisition of minimally invasive surgical skills, demonstrating construct validity. PMID- 15781762 TI - Increased nasal airflow with budesonide compared with desloratadine during the allergy season. AB - OBJECTIVE: To compare the effects of desloratadine, an H1-blocking antihistamine, and budesonide, an intranasal corticosteroid, on nasal peak inspiratory flow (NPIF) in patients with seasonal allergic rhinitis. DESIGN: We performed a randomized, double-blind, double-dummy, parallel study comparing oral desloratadine, 5 mg/d (n = 31), and budesonide, 32 mug/d per nostril (n = 30), for 2 weeks during the spring allergy season. MAIN OUTCOME MEASURES: Subjects recorded NPIF and nasal symptoms twice daily. Baseline measurements were obtained before initiation of treatment. The Rhinoconjunctivitis Quality of Life Questionnaire was completed at baseline and after treatment. RESULTS: Desloratadine and budesonide caused a significant increase in NPIF compared with baseline on the evening of the first dose (P < .01). Budesonide, however, led to a significantly greater increase in NPIF than did desloratadine when the change from baseline was compared for the entire treatment period (median, 475 vs 150 L/min; P = .01). Both treatments resulted in clinically significant reductions of the individual domains and overall scores on the Rhinoconjunctivitis Quality of Life Questionnaire (P < .01). There was a significant reduction in total symptom scores (P < or = .01) compared with baseline during all treatment days in both treatment groups, with no statistically significant differences between treatments (median, -60.0 vs -59.5; P = .67). CONCLUSIONS: Both treatments led to significant improvements in NPIF, but the improvement was greater with the intranasal corticosteroid. Both treatments improved quality of life and reduced symptoms. The difference between the objective and subjective outcomes probably reflects the small sample size, the low pollen counts for the season, and the greater variability in subjective compared with objective measures. PMID- 15781763 TI - Multiplanar computed tomographic analysis of frontal recess cells: effect on frontal isthmus size and frontal sinusitis. AB - BACKGROUND: Frontal recess anatomy can be very complex, with accessory cells such as frontal, agger nasi, and intersinus septal cells encroaching on the frontal recess and possibly contributing to obstruction of the frontal sinus. In this study, we determined the prevalence of these cells and their relationship to frontal sinusitis in patients who have (revision group) and have not (primary group) had previous sinus surgery. DESIGN: Multiplanar computed tomographic images were reconstructed on a computer workstation to determine the presence of frontal, agger nasi, and intersinus septal cells and frontal sinusitis. We also measured the diameter and area of the frontal isthmus for each sinonasal cavity. We were able to retrieve 106 of 117 images from a surgical database encompassing the previous 2 years. SETTING: Tertiary care academic practice of the senior author. RESULTS: Frontal cells were found in 25.5% of frontal recesses, including 29.6% of sides in the primary group and 21.9% of sides in the revision group. We identified 33.0% of patients as having unilateral or bilateral frontal cells. Type I cells were the most common cell (18.4% of primary sinuses). The presence of frontal sinusitis and the diameter and area of the frontal isthmus were not significantly different for those patients with compared with patients without frontal cells. Intact agger nasi cells were identified in 86.7% of primary sinuses and 53.5% of revision sinuses. There was no increased incidence of frontal sinusitis in patients with persistent agger nasi cells in the revision group. CONCLUSIONS: When we evaluated multiplanar reconstructions, we identified frontal cells in 33.0% of patients overall, which was more common than previously reported. The findings of agger nasi cells indicated that these cells were likely addressed in less than half of previous sinus procedures. However, frontal cells and retained agger nasi cells were not associated with a higher incidence of frontal sinusitis, and there was no association between the size of the frontal isthmus and the presence of frontal sinusitis. Although anatomic variations in the frontal recess are likely to play a role in frontal sinusitis, mucosal inflammatory processes are likely to be a much more important etiologic factor. PMID- 15781764 TI - Increased prevalence of chronic rhinosinusitis in carriers of a cystic fibrosis mutation. AB - OBJECTIVE: To explore whether there is an increased prevalence of chronic rhinosinusitis (CRS) in known cystic fibrosis (CF) carriers. Self-reported CRS affects 13% to 14% of the US population and clusters in families, which suggests that genetic factors may play an etiologic role. Cystic fibrosis is an inherited recessive disorder that invariably affects the sinuses. The frequency of CF mutations has been reported to be higher in patients with CRS than in unaffected controls. PATIENTS: Obligate CF carriers (parents of patients with CF) were recruited from the Johns Hopkins CF clinic. The presence of signs and symptoms of CRS was assessed by a sinus disease questionnaire. A subgroup of participants was evaluated by a physician experienced in the diagnosis of CRS. RESULTS: Fifty three (36%) of 147 obligate CF carriers who returned a completed questionnaire had self-reported CRS. Twenty-three CF carriers (14 with and 9 without CRS based on self-reporting in the questionnaire) were clinically evaluated. Seven were diagnosed as having CRS (all 7 with self-reported CRS), while another 6 had allergic rhinitis or recurrent acute rhinosinusitis (all 6 with self-reported CRS), and 10 had no evidence of active sinus disease (1 with self-reported CRS). The sensitivity (100%) and specificity (56%) of the questionnaire for physician diagnosed CRS was similar to that of other survey instruments used to estimate the prevalence of self-reported CRS in the general population. CONCLUSION: Carriers of a single CF mutation have a higher prevalence of CRS than the general population. PMID- 15781765 TI - The prevalence of "incidental" acoustic neuroma. AB - OBJECTIVE: To estimate the prevalence of "incidental" acoustic neuromas (ANs) in the population at large. DESIGN: An intracranial magnetic resonance imaging (MRI) database of 46 414 patients presenting to the University of California, San Francisco (UCSF), without known audiovestibular complaints was searched retrospectively from July 1995 to February 2003. Seventy percent of these MRIs included gadolinium, and none was specifically targeted through the internal auditory canal. A medical chart review of 688 patients with acoustic neuromas presenting to UCSF between 1980 and 1999 was searched for sex distribution. SETTING: Tertiary care university medical center. RESULTS: Eight patients with incidental AN were discovered. This figure suggests that undiagnosed ANs may be present in at least 0.02% of the population. Three patients were found to have audiovestibular symptoms on inquiry after diagnosis. Audiometry revealed asymmetry at 4 kHz in only 3 of 7 patients, with an otherwise symmetric audiogram in the remaining patients. Tumor size in this population ranged from 3 to 28 mm. Incidental ANs were more common in men, but ANs were more common in women overall. CONCLUSIONS: The prevalence of incidental AN appears to be roughly 2 in 10,000 people. This figure indicates that AN may be less prevalent than suggested in previously reported temporal bone studies and more prevalent than suggested by epidemiologic studies. PMID- 15781766 TI - Cochlear implant complications: utility of federal database in systematic analysis. AB - OBJECTIVES: To explore the suitability of the Manufacturer User Facility and Distributor Experience (MAUDE) database (which is maintained by the Food and Drug Administration and has a mandatory reporting requirement) for systemic analysis of cochlear implant complications and treatments and, in so doing, analyze trends in cochlear implant complications for 2 periods, 2002 and pre-1998. DATA SOURCES: All events from 2002 and from before 1998 were considered. Events and action taken were categorized and tabulated. DATA SYNTHESIS: Because there was no null hypothesis, statistical analysis (chi2) was only used in comparing the 2 time frames. CONCLUSIONS: Structural limitations of the database, in addition to disparate reporting quality, made systematic analysis difficult. It was noted that spontaneous device failure accounted for the greatest single number of adverse events for both 2002 and the pre-1998 period (267/654 [41%] and 74/129 [57%], respectively), confirming earlier studies. A statistically significant decrease in spontaneous device failure and a significant increase in infections from the pre-1998 period to 2002 was observed. Flap problems ranging from extrusion to infection that required explantation were less frequently reimplanted than other problems requiring explantation, such as device failure or trauma. We considered new directions, including close collaboration with the new MedSun reporting system and conclude that while a valuable resource for narrative data, the current structure of the MAUDE database is only modestly useful for analyzing trends in complications and cannot answer several crucial questions, including device type comparisons. We suggest changing the current report format to include patient age, duration of implant, presence of anatomical abnormalities, and details on spontaneous device failures. PMID- 15781767 TI - The utility of the MAUDE database in researching cochlear implantation complications. PMID- 15781768 TI - Rates and risk factors for subsequent tonsillectomy after prior adenoidectomy: a regression analysis. AB - OBJECTIVE: To determine the role of adenoidectomy without concurrent tonsillectomy in the treatment of upper airway obstruction, by determining rates and risk factors for subsequent tonsillectomy. DESIGN: Retrospective cohort study with nested case-control study. Data were evaluated using Kaplan-Meyer curves with Cox proportional hazards regression analysis, as well as contingency table and logistic regression analysis. SETTING: Tertiary care pediatric hospital with satellite clinics and surgical centers. PATIENTS: A total of 2462 patients aged 5 months to 18 years undergoing adenoidectomy without concurrent tonsillectomy. MAIN OUTCOME MEASURES: A 5-year database was searched for birth dates, dates of initial surgery, and dates of subsequent tonsillectomy (if performed) or latest follow-up. Cases (tonsillectomies) were then matched 1:1 by age with controls (no subsequent tonsillectomy). Medical charts were reviewed to identify potential predisposing factors, including sex, tonsil size, and adenoidectomy or tonsillectomy indication. RESULTS: Within 5.4 years, 108 patients underwent subsequent tonsillectomy. The relative risk of subsequent tonsillectomy decreases by 0.83 (95% confidence interval, 0.78-0.88) for each increasing year of age at adenoidectomy. The odds of undergoing a future tonsillectomy significantly increase with increasing tonsil size at the time of adenoidectomy. There was a trend toward doubling the risk of subsequent tonsillectomy when the adenoids were removed for upper airway obstruction (including obstructive sleep apnea) compared with other indications (P = .06). CONCLUSION: Knowledge of the rates and risk factors for subsequent tonsillectomy will allow more informed counseling of parents regarding whether tonsillectomy should be performed or deferred at the time of an indicated adenoidectomy. PMID- 15781769 TI - Helicobacter pylori in children who are prone to upper respiratory tract infections. AB - OBJECTIVE: To examine the presence of Helicobacter pylori infection in children with recurrent upper respiratory infections, including acute otitis media. DESIGN: A prospective clinical trial. SETTING: Tertiary referral center. PATIENTS: A total of 20 children who were prone to upper respiratory infections and acute otitis and who were admitted for elective adenoidectomy and/or tympanostomy were included in the study. MAIN OUTCOME MEASURES: Samples of adenoid tissue and middle ear fluid were cultured for H pylori. Serologic analysis and fecal antigen detection testing were also carried out to determine the presence of H pylori. RESULTS: The results of serologic and antigen detection tests were positive for H pylori in 4 children (20%), but cultures of adenoid tissue and middle ear fluid samples were negative for H pylori in all children. CONCLUSION: An association between H pylori infection and recurrent upper respiratory infections and otitis media could not be established. PMID- 15781770 TI - Bilateral obstructing laryngeal epithelial adenomatous hamartomas. PMID- 15781771 TI - Thyroid cartilage fracture: an unusual presentation. PMID- 15781773 TI - Use of SLSE after endoscopic sinus surgery in children should be strictly limited. PMID- 15781772 TI - The pros and cons of second-look sinonasal endoscopy after endoscopic sinus surgery in children. PMID- 15781774 TI - Pathology quiz case: Ossifying fibroma of the maxilla presenting with associated sinusitis and orbital complications. PMID- 15781775 TI - Radiology quiz case 1: Brown tumor of the mandible. PMID- 15781776 TI - Radiology quiz case 2: Thymic remnant. PMID- 15781778 TI - Tonsillectomy techniques. PMID- 15781780 TI - Cordectomy with imbrication laryngoplasty. PMID- 15781781 TI - Greenfield sluder and subtotal tonsillectomy. PMID- 15781782 TI - The Department of Surgery at the University of Cincinnati College of Medicine, Cincinnati, Ohio. PMID- 15781783 TI - Professionalism and the shift mentality: how to reconcile patient ownership with limited work hours. PMID- 15781784 TI - Surgical evolution: collaboration is the key. PMID- 15781785 TI - Women in surgery: the same, yet different. PMID- 15781786 TI - A century of healing, a century of change. PMID- 15781787 TI - A major challenge for graduate medical education. PMID- 15781788 TI - A young surgeon's perspective. AB - The environment of health care delivery is responding to a variety of external forces that are changing the practice of surgery. Only when we look back at this period can any assessments be made to determine whether the changes were beneficial or detrimental. The observation that surgeons are retiring at an earlier age may be a reflection of the negative effect of the changing culture of surgery on career satisfaction. As a young surgeon who anticipates an additional 20+ years of a surgical career, these changes are important to me and will clearly affect my practice as well as my satisfaction. This special article provides my personal perspective on some of the current and anticipated changes in surgery that are a manifestation of cultural changes in health care delivery. PMID- 15781789 TI - The changing roles of a surgical department chair: adapting to a changing environment. PMID- 15781790 TI - The new paradigm of continuing education in surgery. AB - Concerns regarding the shortcomings of traditional continuing medical education (CME) have led to widespread acceptance of the concept of continuous professional development (CPD). Continuous professional development focuses on the individual learning needs of physicians across the continuum of their professional careers and encompasses a broad range of educational activities. Continuous professional development activities of practicing surgeons should be integrated with the core competency of practice-based learning and improvement (PBLI), which involves a cycle of 4 steps-identifying areas for improvement, engaging in learning, applying new knowledge and skills to practice, and checking for improvement. The effectiveness of CPD and PBLI should be objectively evaluated by assessing their impact on surgeons' learning and performance and on patient care outcomes. A portfolio may be used to document CPD and PBLI activities and can serve as a valuable learning and assessment tool. It should contain information on the personal learning projects pursued by the surgeon. Continuous professional development of the entire surgical team and education of patients and their families need to be considered within the context of the surgeon's CPD efforts to positively affect patient care. Thus, a concept of global 360 degrees education should be embraced. Continuous professional development integrated with PBLI can help surgeons address their specific learning needs and play a pivotal role in surgeons' providing the best care to patients. PMID- 15781791 TI - Joseph E. Murray (1919- ): Nobel Laureate, 1990. PMID- 15781793 TI - Postoperative mortality and morbidity in French patients undergoing colorectal surgery: results of a prospective multicenter study. AB - HYPOTHESIS: Better knowledge of independent risk factors might decrease mortality and morbidity rates following colorectal surgery. DESIGN: Prospective multicenter study. INTERVENTIONS: From June to September 2002, consecutive patients undergoing open or laparoscopic surgery (electively or on an emergent basis) for colorectal cancers or diverticular disease were prospectively included. Exclusion criteria were colectomy for other causes (eg, inflammatory bowel diseases, benign polyps). The structured sheet of data collection included more than 200 items on all perioperative data concerning the patient, the disease, and the operating surgeons. Postoperative mortality and morbidity were defined as in-hospital death and complications. RESULTS: Among 1421 patients, the in-hospital death rate was 3.4% and the overall morbidity rate was 35%. Four independent preoperative risk factors of mortality were found: emergency surgery, loss of more than 10% of weight, neurological comorbidity, and age older than 70 years. Six independent risk factors of morbidity were found: age older than 70 years, neurologic comorbidity, hypoalbuminemia, cardiorespiratory comorbidity, long duration of operation, and peritoneal contamination. CONCLUSION: Colorectal resection in France is associated with a 3.4% mortality rate and a 35% morbidity rate. Knowledge of the risk factors could help surgeons manage cases. PMID- 15781792 TI - Impact of donor, technical, and recipient risk factors on survival and quality of life after liver transplantation. AB - HYPOTHESIS: Donor, technical, and recipient risk factors cumulatively impact survival and health-related quality of life after liver transplantation. DESIGN: Retrospective study. SETTING: Tertiary care center. PATIENTS: A total of 483 adults undergoing primary orthotopic liver transplantation between January 1, 1991, and July 31, 2003. MAIN OUTCOME MEASURES: Graft and patient survival, Karnofsky functional performance scores, Medical Outcomes Study Short Form 36 Health Survey scores, and Psychosocial Adjustment to Illness Scale scores as influenced by potential risk factors including donor age, weight, warm ischemia time, cold ischemia time (CIT), sex, United Network for Organ Sharing (UNOS) status (1 or 2A vs 2B or 3), recipient age and disease, bilirubin level, and creatinine level. RESULTS: Five-year graft survival was 72% for recipients of donors younger than 60 years and 35% for recipients of donors 60 years and older (P<.001). A CIT of 12 hours or more was associated with shorter 5-year graft survival (71% vs 58%; P = .004). Five-year graft survival for UNOS status 2B or 3 was 71% vs 60% for status 1 or 2A (P = .02). A comparable pattern was seen for patient survival in relation to donor age (P = .003), CIT (P = .005), and urgency status (P = .03). Urgent UNOS status, advanced donor age, and prolonged CIT were independently associated with shorter graft and patient survival (P<.05). Functional performance and health-related quality of life were not affected by donor, recipient, or technical characteristics. CONCLUSIONS: Combining advanced donor age, urgent status, and prolonged CIT adversely affects graft and patient survival, and the cumulative effects of these risk factors can be modeled to predict posttransplant survival. PMID- 15781794 TI - Is mechanical bowel preparation mandatory for elective colon surgery? A prospective randomized study. AB - BACKGROUND: Bowel preparation prior to colonic surgery usually includes antibiotic therapy together with mechanical bowel preparation (MBP). Mechanical bowel preparation may cause discomfort to the patient, prolonged hospitalization, and water and electrolyte imbalance. It was assumed that with the improvement in surgical technique together with the use of more effective prophylactic antibiotics, it was possible that MBP would no longer be necessary. HYPOTHESIS: There is no statistical difference in the postoperative results of patients who undergo elective colon resection with MBP as compared with those who have no MBP. DESIGN AND PATIENTS: The study includes all patients who had elective large bowel resection at Campus Golda between April 1, 1999, and March 31, 2002. Emergency operations were not included. The patients were randomly assigned to the 2 study groups (with or without MBP) according to identification numbers. All patients were treated with intravenous and oral antibiotics prior to surgery. The patients in the MBP group received Soffodex for bowel preparation. RESULTS: A total of 329 patients participated in the study, 165 without MBP and 164 with MBP. The 2 groups were similar in age, sex, and type of surgical procedure. Two hundred sixty-eight patients (81.5%) underwent surgery owing to colorectal cancer and 61 patients (18.5%) owing to benign disease. The hospitalization period was longer in the bowel-prepared group (mean +/- SD, 8.2 +/- 5.1 days) as compared with the nonprepared group (mean +/- SD, 8.0 +/- 2.7 days). However, this difference was not statistically significant. The time until the first bowel movement was similar between the 2 groups: a mean +/- SD of 4.2 +/- 1.3 days in the nonprepared group as compared with a mean +/- SD of 4.3 +/- 1.1 days in the prepared group (P = NS). Four patients (1.2%) died in the postoperative course owing to acute myocardial infarction and pulmonary embolism. Sixty-two patients (37.6%) of the non-MBP group suffered from postoperative complications as compared with 77 patients (46.9%) of the MBP group. CONCLUSION: Our results suggest that no advantage is gained by preoperative MBP in elective colorectal surgery. PMID- 15781795 TI - Hemodynamic changes during laparoscopic gastric bypass procedures. AB - HYPOTHESIS: Significant detrimental intra-operative hemodynamic and respiratory changes occur in the morbidly obese during laparoscopic gastric bypass. DESIGN: Case series. SETTING: Tertiary care university hospital. PATIENTS: Thirteen patients, 10 women and 3 men, undergoing uncomplicated laparoscopic gastric bypass for morbid obesity. INTERVENTIONS: Using a pulmonary artery catheter and an arterial line, we intraoperatively monitored hemodynamic and respiratory parameters. Parameter values were recorded at set points of the procedure, and the changes were statistically analyzed. RESULTS: Significant hemodynamic and respiratory changes, mostly unfavorable, occur in the morbidly obese when creating the pneumoperitoneum in preparation for laparoscopic gastric bypass. The hemodynamic changes are attenuated when the patient is placed in the reverse Trendelenburg position and almost completely corrected when the abdomen is deflated at the completion of the procedure. The respiratory changes are more persistent. CONCLUSIONS: Laparoscopic gastric bypass surgery for morbid obesity leads to a number of predominantly detrimental, if temporary, respiratory and hemodynamic changes, which are most pronounced at the time of creation of the pneumoperitoneum. In the presence of significant cardiopulmonary comorbidities, the use of invasive intra-operative hemodynamic monitoring of the morbidly obese undergoing laparoscopic gastric bypass appears therefore justified. PMID- 15781796 TI - Synchronized video and motion analysis for the assessment of procedures in the operating theater. AB - HYPOTHESIS: Objective assessment of surgical skill has recently been shown to be possible through the use of dexterity-based and video analysis systems. The aim of this study was to synchronize these 2 modalities to produce a comprehensive surgical assessment tool. DESIGN: The Imperial College Surgical Assessment Device is a dexterity-based motion analysis device that has been developed in the Department of Surgical Oncology and Technology by the Surgical Computing and Imaging Research Group. Further advances to this system have been made to enable synchronized acquisition of hand kinematics and video from real procedures, and their concurrent analysis. To test the feasibility of the system, 10 laparoscopic cholecystectomies performed by 5 different surgeons on consenting patients were recorded. Analysis focused on the entire procedure and also on specific parts of the operation such as the clipping and cutting of the cystic duct and artery. RESULTS: Dexterity analysis was performed using the objective measures of time, path length, number of movements, velocities, and trajectories. Comparative analysis of a surgeon's dexterity was carried out on the whole procedure and by using the synchronized zoom facility in the software. Kinematic signals revealed rapid changes in velocity caused by alternating between different instruments or occurring after complications such as bleeding. CONCLUSION: This new motion analysis system has been shown to be an effective tool for the comprehensive assessment of operative procedures. PMID- 15781797 TI - Medical and surgical management of chronic ulcerative colitis. PMID- 15781798 TI - Image of the month. Papillary low-grade neuroendocrine tumor of the pancreas with pancreatic duct dilation. PMID- 15781799 TI - Loyal Davis' honorary fellowship. PMID- 15781800 TI - Bilateral chorea-ballism after cardiac arrest. PMID- 15781801 TI - Cerebellar agenesis. PMID- 15781802 TI - Patient page. Exercise is safe and beneficial for people with facioscapulohumeral muscular dystrophy. PMID- 15781803 TI - What can we say to women of reproductive age with epilepsy? PMID- 15781804 TI - Optic mitochondriopathies. PMID- 15781805 TI - Specialty care by child neurologists: a workforce analysis. AB - OBJECTIVE: To provide a current profile of the practice of child neurology, report the attitudes of child neurologists toward practice, and analyze the supply of child neurologists. METHODS: In March 2002, a questionnaire was sent to all active members of the Child Neurology Society (n = 1,051) and to nonmember physicians under age 70 who listed child neurology as a primary or secondary specialty on the American Medical Association Masterfile (n = 433). The response rate was 65%. Eligibility criteria were then applied to arrive at the sample of main specialty in child neurology working at least 20 hours per week in patient care. The final population was 604. Differences in practice characteristics were tested by practice type, and the number of full-time patient care child neurologists was projected by extrapolating to nonrespondents. RESULTS: There are 904 full-time patient care child neurologists in the United States and 1.27 per 100,000 children. Career satisfaction is 90%, yet no growth in the supply is projected over the next 20 years. Wait times for an appointment average 53 and 44 days for a new and return visit, with longer wait times in university settings. Average annual income is 151,000 dollars. CONCLUSION: The practice characteristics of child neurologists suggest that the specialty will be challenged to meet patient demands. PMID- 15781806 TI - Neuropsychological effects of exposure to anticonvulsant medication in utero. AB - OBJECTIVE: To investigate the long-term differential drug effects on cognitive functioning in school-aged children exposed to antiepileptic drugs (AEDs) in utero. METHODS: Mothers with epilepsy were recruited from specialist epilepsy clinics and obstetric clinics from the Liverpool and Manchester region. The mothers and their children were recruited without prior knowledge of their AED treatment during pregnancy or the health of the offspring. A battery of neuropsychological tests was applied to each mother-child pair in order to obtain a neuropsychological profile for each child. RESULTS: Neuropsychological investigation was performed on 249 children between the ages of 6 and 16. Children exposed to sodium valproate had a significantly lower verbal IQ when compared to children exposed to other antiepileptic drugs or not exposed at all. The same children were more likely to have an IQ below 69 and more likely to have memory impairment when compared to the other groups. The mothers' IQ, exposure to sodium valproate, and the number of tonic-clonic seizures during pregnancy were significant predictors of verbal IQ in this population. CONCLUSIONS: This retrospective study highlights the potential harmful effects of sodium valproate exposure in utero on neuropsychological development. PMID- 15781807 TI - Lamotrigine and the risk of malformations in pregnancy. AB - OBJECTIVE: To report the frequency of major malformations in lamotrigine-exposed pregnancies from September 1, 1992, through March 31, 2004, in the International Lamotrigine Pregnancy Registry. METHODS: Health care professionals throughout the world can voluntarily enroll lamotrigine-exposed pregnancies in this observational study. Only pregnancies with unknown outcomes at the time of enrollment were included in the analysis. The percentage of outcomes with major birth defects was calculated as the total number of outcomes with major birth defects divided by the sum of the number of outcomes with major birth defects + the number of live births without defects. RESULTS: Among 414 first-trimester exposures to lamotrigine monotherapy, 12 outcomes with major birth defects were reported (2.9%, 95% CI 1.6% to 5.1%). Among the 88 first-trimester exposures to lamotrigine polytherapy including valproate, 11 outcomes with major birth defects were reported (12.5%; 95% CI 6.7% to 21.7%). Among 182 first-trimester exposures to lamotrigine polytherapy excluding valproate, 5 outcomes with major birth defects were reported (2.7%, 95% CI 1.0% to 6.6%). No distinctive pattern of major birth defects was apparent among the offspring exposed to lamotrigine monotherapy or polytherapy. CONCLUSIONS: The risk of all major birth defects after first-trimester exposure to lamotrigine monotherapy (2.9%) was similar to that in the general population and in other registries enrolling women exposed to antiepileptic monotherapy (3.3% to 4.5%). However, the sample size was too small to detect any but very large increases in specific birth defects. PMID- 15781808 TI - Increased rate of major malformations in offspring exposed to valproate during pregnancy. AB - OBJECTIVE: To determine the rate of occurrence of major malformations in infants whose mothers had taken the drug valproic acid (VPA) as monotherapy during the first trimester of pregnancy and had enrolled in the North American Antiepileptic Drug Pregnancy Registry. METHODS: Data were collected from pregnant women throughout the United States and Canada through telephone-based interviews. Each woman was interviewed at enrollment, at 7 months' gestation, and postpartum. With her written permission, the medical records of each mother and her infant were obtained. The major malformations tabulated were those identified at or before 5 days of age. The prevalence of congenital malformations among offspring of monotherapy VPA-exposed women was compared with that among infants of women exposed to all other antiepileptic drugs (internal comparison group) and with that among newborns in the Active Malformations Surveillance Program at Brigham and Women's Hospital (external comparison group). RESULTS: Sixteen affected cases were identified among 149 VPA-exposed women (proportion: 10.7%; 95% CI: 6.3 to 16.9%). The prevalence in the internal comparison group was 2.9% (95% CI: 2.0 to 4.1%; odds ratio: 4.0, 95% CI: 2.1 to 7.4; p < 0.001). Assuming a 1.62% prevalence in the external comparison group, the relative risk of having an affected offspring for VPA-exposed women was 7.3 (95% CI: 4.4 to 12.2; p < 0.001). CONCLUSION: Maternal exposure to valproic acid during the first trimester of pregnancy significantly increased the risk of major malformations. PMID- 15781809 TI - Mitochondrial DNA content is decreased in autosomal dominant optic atrophy. AB - BACKGROUND: Autosomal dominant optic atrophy (ADOA) is the commonest form of inherited optic neuropathy. Mutations in the OPA1 gene encoding a dynamin-related mitochondrial protein underlie ADOA and may perturb the biogenesis and maintenance of mitochondria. OBJECTIVE: To investigate the mutation spectrum of the OPA1 gene and assess alterations in mitochondrial content caused by OPA1 mutations. METHODS: Sixteen Korean patients with clinically suspected ADOA were studied. The mutation spectrum of the OPA1 gene was analyzed by PCR single-strand conformation polymorphism and sequencing, and mitochondrial DNA (mtDNA) content was quantified by real-time PCR. RESULTS: Eight different mutations were found, including five novel mutations. Quantitative real-time PCR analysis showed excellent linearity and precision for the determination of mtDNA copy numbers. The number of mtDNA copies per cell in patients with OPA1 gene mutations (ages 7 to 40) was significantly lower than those in all normal control subjects (p = 0.037), particularly lower than in normal control subjects ages 10 to 39 (p = 0.022). CONCLUSION: The mutation spectrum of the OPA1 gene disclosed marked genetic heterogeneity and the mitochondrial DNA content was found to be lower in autosomal dominant optic neuropathy, which provides direct evidence for a pathogenetic role of mutations of the OPA1 gene. PMID- 15781810 TI - The frequency of intractable seizures after stopping AEDs in seizure-free children with epilepsy. AB - BACKGROUND: After 1 to 4 years, seizure-free children with epilepsy are encouraged to stop daily antiepileptic drug (AED) treatment. Approximately 70% are successful. The authors examined how often intractable epilepsy follows discontinuation of AED treatment in a population-based cohort of children with epilepsy. METHODS: The Nova Scotia population-based epilepsy cohort was used to identify children who discontinued AEDs but subsequently developed intractable epilepsy. All patients studied (ages 1 month to 16 years) developed epilepsy between 1977 and 1985, had epilepsies characterized by partial or convulsive seizures, and had at least 5 years of follow-up evaluation (n = 367). Those with benign rolandic epilepsy were excluded. Intractability was defined as one or more seizures every 3 months during the last year of follow-up review or until successful seizure surgery and failure of three or more AEDs at maximum tolerated doses. RESULTS: Overall, 71% (260/367) of eligible children became free of seizure for 1 to 4 years and discontinued AED treatment. Of this group, 70% remained seizure-free without AED treatment, but 30% had recurrences. Only three children with recurrences later developed intractable epilepsy. Two then underwent a temporal lobectomy, one successful and one only partially successful (20-year follow-up periods). The third patient continued to have intractable epilepsy for 7 years after discontinuing AED treatment but eventually entered remission. CONCLUSION: Approximately 1% of children who became free of seizure and discontinued antiepileptic drug treatment had recurrent seizures that could not be controlled again with medication. The authors were unable to predict this outcome. It remains unclear whether a similar outcome would have occurred if antiepileptic drugs had not been discontinued. PMID- 15781811 TI - Prevalence of large-scale mitochondrial DNA deletions in an adult Finnish population. AB - BACKGROUND: Large-scale mitochondrial DNA (mtDNA) deletions are associated with clinical conditions such as Kearns-Sayre syndrome and chronic progressive external ophthalmoplegia in adults and Pearson syndrome in children. Reported case series have suggested that deletions are not uncommon in the population, but their prevalence has not been documented. METHODS: The authors ascertained patients with clinical features associated with mtDNA deletions in a defined adult population in northern Finland. Buccal epithelial samples were requested from each patient fulfilling the selection criteria, and full-length mtDNA was amplified using the long PCR method. Deletion breakpoints were identified using sequencing. Patients with deletions were examined clinically. RESULTS: The authors identified four patients with single large-scale mtDNA deletions. The prevalence of deletions was calculated to be 1.6/100,000 in the adult population in the province of Northern Ostrobothnia (0.0 to 3.2; 95% CI). Analysis of incident cases from a neighboring province revealed two patients with deletions and yielded a similar population frequency. CONCLUSIONS: The frequency of large scale mitochondrial DNA deletions is similar among populations, suggesting that there is a constant rate of new deletions. PMID- 15781812 TI - Lafora disease due to EPM2B mutations: a clinical and genetic study. AB - OBJECTIVE: To study EPM2B gene mutations and genotype-phenotype correlations in patients with Lafora disease. METHODS: The authors performed a clinical and mutational analysis of 25 patients, from 23 families, diagnosed with Lafora disease who had not shown mutations in the EPM2A gene. RESULTS: The authors identified 18 mutations in EPM2B, including 12 novel mutations: 4 nonsense mutations (R265X, C26X, W219X, and E67X), a 6-base pair (bp) microdeletion resulting in a two amino acid deletion (V294_K295del), a 4-bp insertion resulting in a frameshift mutation (S339fs12), and 6 missense mutations (D308A, I198N, C68Y, E67Q, P264H, and D233A). In our data set of 77 families with Lafora disease, 54 (70.1%) tested probands have mutations in EPM2A, 21 (27.3%) in EPM2B, and 2 (2.6%) have no mutations in either gene. The course of the disease was longer in patients with EPM2B mutations vs patients with EPM2A mutations. CONCLUSIONS: Genetic allelic heterogeneity is present in Lafora disease associated with mutations in EPM2B. Patients with mutations in EPM2A and EPM2B express similar clinical manifestation, although patients with EPM2B-associated Lafora disease seem to have a slightly milder clinical course. The lack of mutations in EPM2A and EPM2B in two families could be because of the presence of mutations in noncoding, nontested regions or the existence of an additional gene associated with Lafora disease. PMID- 15781813 TI - Treatment with laquinimod reduces development of active MRI lesions in relapsing MS. AB - BACKGROUND: Laquinimod is a novel immunomodulatory substance developed as an orally available disease modifying treatment in multiple sclerosis (MS). The purpose of this study was to evaluate safety, tolerability, and efficacy on MRI lesions of two different doses of laquinimod compared with placebo in patients with relapsing MS. METHODS: In this multicenter, double-blind, randomized trial, patients with relapsing MS received 0.1 mg or 0.3 mg laquinimod or placebo as three daily tablets for 24 weeks. Gadolinium-enhanced brain MRI scans were performed at screening, every eighth week during treatment, and 8 weeks after end of treatment. The primary efficacy variable was the cumulative number of active lesions over 24 weeks. Safety measures included adverse events, physical examination, and laboratory variables. RESULTS: Of 256 screened patients, 209 were randomized (67 to 74 patients per group) in 20 centers. There was a significant difference between laquinimod 0.3 mg and placebo for the primary outcome measure (mean cumulative number of active lesions reduced by 44%). In the subgroup of patients with at least one active lesion at baseline the reduction was slightly more pronounced (52%). No differences with respect to clinical variables (relapses, disability) were found. The safety profile was favorable; there were no clinical signs of undesired inflammatory manifestations. CONCLUSION: Oral laquinimod in a dosage of 0.3 mg daily was well tolerated and effective in suppressing development of active lesions in relapsing multiple sclerosis. PMID- 15781814 TI - Low-contrast letter acuity testing captures visual dysfunction in patients with multiple sclerosis. AB - OBJECTIVE: To evaluate concurrent and predictive validity for low-contrast letter acuity (L-CLA) testing as a candidate visual component for the Multiple Sclerosis Functional Composite (MSFC). METHODS: L-CLA testing was conducted in two MS patient cohorts. In the MSFC Validation Study, 137 participants from a Phase III trial of inteferon beta-1a (Avonex) for relapsing-remitting MS were followed. A second cohort included 65 patients with secondary progressive MS who participated in a substudy of the International MS Secondary Progressive Avonex Controlled Trial (IMPACT). The total number of letters read correctly at four contrast levels (100, 5, 1.25, and 0.6%) was correlated with Expanded Disability Status Scale (EDSS), MSFC, Sickness Impact Profile, Multiple Sclerosis Quality of Life Inventory, and brain parenchymal fraction (BPF), as determined by MRI. RESULTS: Low- and high-contrast letter acuity scores correlated with BPF at follow-up in the MSFC Validation Study (5%: r = 0.40, p < 0.0001; 100%: r = 0.31, p = 0.0002). L-CLA also correlated with EDSS (5%: r = -0.35, p < 0.0001; 1.25%: r = -0.26, p = 0.0003) and MSFC (5%: r = 0.47, p < 0.0001; 1.25%: r = 0.45, p < 0.0001). In the IMPACT Substudy, change in L-CLA scores from baseline to year 1 predicted subsequent change in the EDSS from year 1 to 2 at the 5% (p = 0.0142) and the 1.25% (p = 0.0038) contrast levels, after adjusting for change in MSFC scores from baseline to year 1. CONCLUSIONS: Low-contrast letter acuity (L-CLA) scores demonstrate concurrent and predictive validity in patients with relapsing remitting and secondary progressive multiple sclerosis (MS). L-CLA testing provides additional information relevant to the MS disease process that is not entirely captured by the Multiple Sclerosis Functional Composite. PMID- 15781815 TI - Interferon beta-1b treatment does not induce autoantibodies. AB - BACKGROUND: There is little information regarding the potential of interferon beta (IFNbeta) to induce or exacerbate autoimmune disease. Existing data from uncontrolled studies are contradictory and do not differentiate between autoimmune dysfunction, which is frequent in patients with multiple sclerosis (MS), and untoward drug effects. OBJECTIVE: To evaluate the impact of IFNbeta on hepatic, thyroid, and other markers of autoimmunity using data from the European placebo-controlled double-blind, multicenter study of IFNbeta-1b in patients with secondary progressive MS (SPMS). METHODS: Serum samples obtained at baseline and at 6-month intervals for 24 months were analyzed for the following autoantibodies (AAbs): antinuclear (ANA), antimitochondrial (AMA), smooth muscle (SMA), liver kidney microsome (LKM), thyroid microsome (TPO), and human thyroglobulin (TG). AAb status at baseline and during treatment was related to respective laboratory and clinical deviations. RESULTS: The analysis of AAb data included 355 patients receiving IFNbeta-1b and 353 receiving placebo. There was no difference between treatment groups in de novo AAb positivity. A greater proportion of women were AAb positive at baseline and during treatment. No association was found between liver enzyme elevations and ANA, AMA, or SMA antibody formation in either treatment group. Laboratory-based thyroid alterations during the study were significantly related to TG/TPO status at baseline but were not associated with IFNbeta-1b treatment. Adverse events possibly indicative of other diseases with autoimmune links were not associated with respective AAb status. CONCLUSION: Interferon beta-1b treatment did not induce autoantibody formation in this population of patients with secondary progressive multiple sclerosis. PMID- 15781817 TI - CNS myelomatosis. PMID- 15781816 TI - Gray and white matter volume changes in early RRMS: a 2-year longitudinal study. AB - BACKGROUND: Brain atrophy, in excess of that seen with normal aging, has been observed early in the clinical course of relapsing-remitting multiple sclerosis (RRMS). Previous work has suggested that at this stage of the disease, gray matter (GM) atrophy progresses more rapidly than the white matter (WM) atrophy. OBJECTIVES: To characterize the evolution of GM and WM volumes over 2 years, and their associations with lesion loads in a cohort of patients with clinically early RRMS. METHODS: Twenty-one patients with RRMS (mean age 37.5 years, mean disease duration from symptom onset 2.1 years) and 10 healthy control subjects (mean age 37.1 years) were studied. Tissue volumes, as fractions of total intracranial volumes, were estimated at baseline and 1- and 2-year follow-up. Brain parenchymal fractions (BPF), GM fractions (GMF), and WM fractions (WMF) were estimated. In subjects with MS, brain lesion loads were determined on conventional T2-weighted along with pre- and post-gadolinium (Gd) enhanced T1 weighted images at each timepoint. RESULTS: A decrease in GMF was observed in subjects with MS vs normal controls over the 2 years of the study (mean -2.1% vs 1.0%, p = 0.044), while no change was seen in WMF over the same period (mean 0.09% vs +0.09%, p = 0.812). However, when the MS cohort was divided in half, dependent upon change in Gd-enhancing lesion load over 2 years (n = 20), a decrease in WMF was seen in the group (n = 10) with the largest decline in Gd volume, whereas WMF increased in the other half (n = 10) concurrent with a net increase in volume of Gd-enhancing lesions (difference between groups: p = 0.034). CONCLUSIONS: Increasing gray matter but not white matter (WM) atrophy was observed early in the clinical course of relapsing-remitting multiple sclerosis. Fluctuations in inflammatory WM lesions appear to be related to volume changes in WM over this time period. PMID- 15781818 TI - SSRI and statin use increases the risk for vasospasm after subarachnoid hemorrhage. AB - BACKGROUND: Use of medications with vasoconstrictive or vasodilatory effects can potentially affect the risk for vasospasm after aneurysmal subarachnoid hemorrhage (SAH). METHODS: Using International Classification of Diseases-9 diagnostic codes followed by medical record review, the authors identified 514 patients with SAH admitted between 1995 and 2003 who were evaluated for vasospasm between days 4 and 14. The authors determined risks for vasospasm, symptomatic vasospasm, and poor clinical outcomes in patients with documented pre-hemorrhagic use of calcium channel blockers, beta-receptor blockers, ACE inhibitors, aspirin, selective serotonin reuptake inhibitors (SSRIs), non-SSRI vasoactive antidepressants, or statins. RESULTS: Vasospasm developed in 62%, and symptomatic vasospasm in 29% of the cohort. On univariate analysis, the risk for all vasospasm tended to increase in patients taking SSRIs (p = 0.09) and statins (p = 0.05); SSRI use increased the risk for symptomatic vasospasm (p = 0.028). The Cochran-Armitage trend test showed that the proportion of patients taking SSRIs and statins increased significantly across three worsening categories (none, asymptomatic, symptomatic) of vasospasm. Logistic regression analysis showed that SSRI use tended to predict all vasospasm (O.R. 2.01 [0.91 to 4.45]), and predicted symptomatic vasospasm (O.R. 1.42 [1.06 to 4.33]). Statin exposure increased the risk for vasospasm (O.R. 2.75 [1.16 to 6.50]), perhaps from abrupt statin withdrawal (O.R. 2.54 [0.78 to 8.28]). Age < 50 years, Hunt-Hess grade 4 or 5, and Fisher Group 3 independently predicted all vasospasm, symptomatic vasospasm, poor discharge clinical status, and death. CONCLUSION: Selective serotonin reuptake inhibitor and statin users have a higher risk for subarachnoid hemorrhage-related vasospasm. Whether the underlying disease indication, direct actions, or rebound effects from abrupt drug withdrawal account for the associated risk warrants further investigation. PMID- 15781819 TI - Posterior thalamic hemorrhage induces "pusher syndrome". AB - BACKGROUND: Recent findings argue for a pathway in humans for sensing the orientation of gravity and controlling upright body posture, separate from the one for orientation perception of the visual world. Stroke patients with contraversive pushing were shown to experience their body as oriented upright when actually tilted about 20 degrees to the ipsilesional side, in spite of normal visual-vestibular functioning. A recent study suggested the involvement of posterolateral thalamus typically associated with the disorder. OBJECTIVE: To evaluate the relationship between pushing behavior and thalamic function. METHODS: Over a 3-year period the authors prospectively investigated 40 patients with left- or right-sided thalamic strokes. RESULTS: Twenty-eight percent showed contraversive pushing. The authors found a strong relationship between etiology, vascular territory, lesion size, and neurologic disorders associated with contraversive pushing. Pusher patients had larger lesions that typically were caused by hemorrhage (vs infarcts) located in the posterior thalamus (vs anterior thalamic lesions in those patients without pushing behavior). A paresis of the contralesional extremities was more frequent and more severe in pusher patients. Further, these patients showed more additional spatial neglect with right thalamic lesions, while they tended to be more aphasic with left thalamic lesions. CONCLUSIONS: Posterior thalamus seems to be fundamentally involved in our control of upright body posture. Higher pressure, swelling, and other secondary pathologic processes associated with posterior thalamic hemorrhage (vs thalamic infarction) may provoke contraversive pushing in combination with additional neurologic symptoms. PMID- 15781820 TI - Migraine, headache, and the risk of stroke in women: a prospective study. AB - BACKGROUND: Migraine and headache in general have been associated with subsequent risk of stroke, primarily in retrospective case-control studies. Prospective data evaluating the association between specific headache forms and stroke are sparse. METHODS: A prospective cohort study was conducted among 39,754 US health professionals age 45 and older participating in the Women's Health Study with an average follow-up of 9 years. Incident stroke was self-reported and confirmed by medical record review. RESULTS: A total of 385 strokes (309 ischemic, 72 hemorrhagic, and 4 undefined) occurred. Compared with nonmigraineurs, participants who reported migraine overall or migraine without aura had no increased risk of any stroke type. Participants who reported migraine with aura had increased adjusted hazards ratios (HRs) of 1.53 (95% CI 1.02 to 2.31) for total stroke and 1.71 (95% CI 1.11 to 2.66) for ischemic stroke but no increased risk for hemorrhagic stroke. Participants with migraine with aura who were <55 years old had a greater increase in risk of total (HR 1.75; 95% CI 1.02 to 3.00) and ischemic (HR 2.25; 95% CI 1.30 to 3.91) stroke. Compared with participants without headache, headache in general and nonmigraine headache were not associated with total, ischemic, or hemorrhagic stroke. CONCLUSIONS: In these prospective data, migraine was not associated with total, ischemic, or hemorrhagic stroke. In subgroup analyses, we found increased risks of total and ischemic stroke for migraineurs with aura. The absolute risk increase was, however, low, with 3.8 additional cases per year per 10,000 women. PMID- 15781821 TI - Neuropsychological predictors of dependency in patients with Alzheimer disease. AB - OBJECTIVE: To determine whether specific cognitive deficits can predict the progression of Alzheimer disease (AD). METHODS: Two hundred fifty-two patients with AD enrolled in the Predictors Study were followed at 6-month intervals for up to 4.5 years with neurologic, cognitive, and psychiatric examinations. Neuropsychological functions were assessed by the Modified Mini-Mental State Examination (mMMSE). Items of mMMSE were divided into five cognitive domains: temporospatial orientation, short-term memory, long-term memory, language, and visuoconstructive functions. Loss of autonomy was assessed by both the Dependency Scale (DS) and the Equivalent Institutional Care (EIC) rating. Cox proportional hazards models, adjusted for age, sex, estimated duration of illness at entry into the study, and presence of extrapyramidal signs and behavioral disturbances, were used to determine the predictive value of each neuropsychological domain on dependency outcomes. RESULTS: Global mMMSE, temporospatial orientation, and short term memory scores were associated with a greater relative risk of moderate or severe dependency. The visuoconstructive score predicted the development of severe dependency. Long-term memory and language scores were not predictive of the EIC or DS endpoints. CONCLUSIONS: The presence of certain neuropsychological deficits at a patient's initial visit, such as short-term memory, temporospatial orientation, and constructive apraxia, predict more rapid dependency in patients with Alzheimer disease. Neuropsychological items have different weights in term of predictive power, and these effects are independent of the influence of age and disease duration at baseline. PMID- 15781822 TI - Normative estimates of cross-sectional and longitudinal brain volume decline in aging and AD. AB - OBJECTIVE: To test the hypotheses 1) that whole-brain volume decline begins in early adulthood, 2) that cross-sectional and longitudinal atrophy estimates agree in older, nondemented individuals, and 3) that longitudinal atrophy accelerates in the earliest stages of Alzheimer disease (AD). METHODS: High-resolution, high contrast structural MRIs were obtained from 370 adults (age 18 to 97). Participants over 65 (n = 192) were characterized using the Clinical Dementia Rating (CDR) as either nondemented (CDR 0, n = 94) or with very mild to mild dementia of the Alzheimer type (DAT, CDR 0.5 and 1, n = 98). Of these older participants, 79 belonged to a longitudinal cohort and were imaged again a mean 1.8 years after baseline. Estimates of gray matter (nGM), white matter (nWM), and whole-brain volume (nWBV) normalized for head sizes were generated based on atlas registration and image segmentation. RESULTS: Hierarchical regression of nWBV estimates from nondemented individuals across the adult lifespan revealed a strong linear, moderate quadratic pattern of decline beginning in early adulthood, with later onset of nWM than nGM loss. Whole-brain volume differences were detected by age 30. The cross-sectional atrophy model overlapped with the rates measured longitudinally in older, nondemented individuals (mean decline of 0.45% per year). In those individuals with very mild DAT, atrophy rate more than doubled (-0.98% per year). CONCLUSIONS: Nondemented individuals exhibit a slow rate of whole-brain atrophy from early in adulthood with white-matter loss beginning in middle age; in older adults, the onset of dementia of the Alzheimer type is associated with a markedly accelerated atrophy rate. PMID- 15781823 TI - Excess of serum copper not related to ceruloplasmin in Alzheimer disease. AB - OBJECTIVE: To assess the role of serum copper in relation to ceruloplasmin and other peripheral markers of inflammation in Alzheimer disease (AD). METHODS: The authors studied serum levels of copper, ceruloplasmin, and transferrin, as well as total peroxides, antioxidants, and other peripheral markers of inflammation in 47 patients with AD, 24 patients with vascular dementia (VaD), and 44 healthy controls. Biochemical variables were related to the patients' and controls' clinical status. RESULTS: The authors found that copper (p < 0.001), peroxides (p = 0.026), and ceruloplasmin (p = 0.052) were increased and TRAP was decreased (p = 0.006) in patients with AD, while no other markers of inflammation were altered. The calculation of the ratio between copper and ceruloplasmin suggested the presence in the serum of AD patients, but not of VaD or normal controls, of a large pool of non-ceruloplasmin-bound copper. CONCLUSIONS: Changes in the distribution of the serum copper components, consisting of an increase of a copper fraction not explained by ceruloplasmin, seem to be characteristic of Alzheimer disease and may be implicated in the pathogenesis of the disease. PMID- 15781825 TI - Sonothrombolysis in acute ischemic stroke for patients ineligible for rt-PA. AB - The authors studied the effect of transcranial ultrasound on patients with acute middle cerebral artery occlusion and contraindications for thrombolysis. Fifteen consecutive subjects were randomized for insonation over 1 hour or for inclusion in a control group. By day 4, recanalization and neurologic improvement occurred more frequently in the target group. Ultrasound-induced acceleration of clot dissolution may be an option for patients with contraindications for recombinant tissue plasminogen activator. PMID- 15781824 TI - Consumption of milk and calcium in midlife and the future risk of Parkinson disease. AB - OBJECTIVE: To examine the relation between milk and calcium intake in midlife and the risk of Parkinson disease (PD). METHODS: Findings are based on dietary intake observed from 1965 to 1968 in 7,504 men ages 45 to 68 in the Honolulu Heart Program. Men were followed for 30 years for incident PD. RESULTS: In the course of follow-up, 128 developed PD (7.1/10,000 person-years). Age-adjusted incidence of PD increased with milk intake from 6.9/10,000 person-years in men who consumed no milk to 14.9/10,000 person-years in men who consumed >16 oz/day (p = 0.017). After further adjustment for dietary and other factors, there was a 2.3-fold excess of PD (95% CI 1.3 to 4.1) in the highest intake group (>16 oz/day) vs those who consumed no milk. The effect of milk consumption on PD was also independent of the intake of calcium. Calcium from dairy and nondairy sources had no apparent relation with the risk of PD. CONCLUSIONS: Findings suggest that milk intake is associated with an increased risk of Parkinson disease. Whether observed effects are mediated through nutrients other than calcium or through neurotoxic contaminants warrants further study. PMID- 15781826 TI - Long-term outcome of endovascular stenting for symptomatic basilar artery stenosis. AB - Eighteen patients underwent stenting for symptomatic basilar artery stenosis. There were three major periprocedural complications (16.7%) without fatality. At a mean 26.7 +/- 12.1-month follow-up, 15 patients (83.3%) had an excellent long term outcome. Only one patient (5.6%) had moderate disability from recurrent stroke, and two patients died of medical illness at 30 and 36 months after stenting. In this uncontrolled study, stenting appeared to be effective in reducing stroke risk and death and worthy of further scrupulous trial. PMID- 15781827 TI - Prospective reliability of the STRokE DOC wireless/site independent telemedicine system. AB - The authors evaluated a site-independent telemedicine system. Telemedicine may be limited by the need for fixed connectivity. Wireless and site-independent technologies eliminate this limitation. Twenty-five stroke patients underwent evaluations by remote and bedside examiners. Ten of 15 (67%) NIH Stroke Scale and 9 of 11 (82%) Modified NIH Stroke Scale items showed excellent interrater reliability. Spearman correlations were > or =0.93. This Internet system is reliable and valid. Further studies should assess its use in acute stroke. PMID- 15781829 TI - Aerobic training improves exercise performance in facioscapulohumeral muscular dystrophy. AB - Exercise programs have been shown to increase strength and endurance in patients with myopathic disorders. The authors investigated the effect of aerobic training in patients with facioscapulohumeral dystrophy (FSHD). Twelve weeks of low intense aerobic exercise improved maximal oxygen uptake and workload with no signs of muscle damage. The authors conclude that aerobic training is a safe method to increase exercise performance in patients with FSHD. PMID- 15781828 TI - Correlation of proband and sibling stroke latency: the SWISS Study. AB - The authors found a correlation between the age at which probands experience an incident stroke and the age at which their siblings experience an incident stroke (r = 0.68; p < 0.0001). Proband-sibling incident stroke latency correlations were observed in analyses restricted to siblings concordant for smoking (r = 0.68; p < 0.0001), diabetes (r = 0.73; p < 0.0001), and hypertension (r = 0.63; p < 0.0001). In the authors' cohort of affected sibling pairs, inherited factors were important determinants of incident ischemic stroke latency. PMID- 15781830 TI - Missing nail for 22 years. PMID- 15781831 TI - Recruitment of contralesional motor cortex in stroke patients with recovery of hand function. AB - In neuroimaging studies of stroke patients, coactivation may account for increased recruitment of bilateral motor areas when moving the affected limb. Here we studied eight patients after stroke with fMRI and simultaneous EMG. Bilateral recruitment of premotor and primary motor cortices was evident in five patients with strictly unilateral performance per EMG. Because patients had excellent motor recovery, this increased recruitment suggests an adaptive response to the infarct. PMID- 15781832 TI - A 15-year follow-up of neuromuscular function in patients with prior poliomyelitis. AB - A population-based cohort of poliomyelitis survivors was established and followed for 15 years (mean time since poliomyelitis was 40 years). Over time, the cohort demonstrated only a modest decline in function as measured by strength measurements, electrophysiologic assessments, and timed functional tasks. There was no association between symptoms of late deterioration and magnitude of decline. Rather, the presence of these symptoms was associated with the magnitude of the residual deficits. PMID- 15781833 TI - In situ identification of hepatitis C virus RNA in muscle. AB - The authors examined skeletal muscle specimens from four patients with myositis and hepatitis C virus (HCV) infection. PCR analysis identified HCV RNA in muscle homogenates from two patients. In situ hybridization signals for HCV RNA were detected within muscle fibers as well as in infiltrating lymphocytes from the same patients. The results may relate to the pathomechanism of myositis in patients with HCV infection. PMID- 15781834 TI - Dose-related vincristine-induced peripheral neuropathy with unexpected off therapy worsening. AB - This study describes the natural course of vincristine-induced peripheral neuropathy in patients with lymphoma (n = 114) receiving vincristine in two different dose intensities. Neuropathic changes were observed in both dose intensity groups, but the higher dose intensity group reported significantly more symptoms during therapy, whereas neurologic signs were significantly more prominent after a cumulative dose of 12 mg vincristine. Furthermore, off-therapy worsening of symptoms (24%) and signs (30%) occurred unexpectedly. PMID- 15781835 TI - The effect of levetiracetam on essential tremor. AB - The effect of a single dose of 1,000 mg of levetiracetam on essential tremor was investigated in 24 patients in a double-blind, placebo-controlled trial. There was a significant reduction of hand tremor for at least 2 hours as measured by accelerometry and functional tests. PMID- 15781836 TI - Urinary 8-hydroxydeoxyguanosine levels as a biomarker for progression of Parkinson disease. AB - 8-hydroxydeoxyguanosine (8-OHdG) has been used to evaluate oxidative stress. The authors investigated urinary 8-OHdG levels in 72 patients with Parkinson disease (PD) and in normal and disease control groups. The mean urinary 8-OHdG increased with the stage of PD and was not influenced by the current dose of DOPA. Our results suggest that urinary 8-OHdG is a potentially useful biomarker for evaluating the progression of PD. PMID- 15781837 TI - Levetiracetam for seizures after liver transplantation. AB - Seizures may occur after orthotopic liver transplantation. Antiepileptic drugs (AEDs) are used to treat these seizures, but the immunosuppressant regimen also may be altered. Levetiracetam is an attractive treatment because of its efficacy, lack of hepatic enzyme induction, and its rapid attainment of serum levels. Treatment with levetiracetam is efficacious, and levetiracetam-treated patients require significantly lower doses of immunosuppressant medications to achieve an equivalent antirejection effect. PMID- 15781838 TI - Incidence of seizures in patients with multiple sclerosis treated with intrathecal baclofen. AB - Oral and intrathecal baclofen (ITB) have been associated with epileptic seizures. The authors observed a higher incidence of epileptic seizures in 99 patients with multiple sclerosis (MS) treated with ITB vs a matched control group (7% vs 1%, p < 0.05). Three patients with MS on ITB developed status epilepticus. Seizures were often associated with additional triggering factors. PMID- 15781839 TI - Cerebral folate deficiency with developmental delay, autism, and response to folinic acid. AB - The authors describe a 6-year-old girl with developmental delay, psychomotor regression, seizures, mental retardation, and autistic features associated with low CSF levels of 5-methyltetrahydrofolate, the biologically active form of folates in CSF and blood. Folate and B12 levels were normal in peripheral tissues, suggesting cerebral folate deficiency. Treatment with folinic acid corrected CSF abnormalities and improved motor skills. PMID- 15781840 TI - Nonsense mutations in mitochondrial DNA associated with myalgia and exercise intolerance. PMID- 15781841 TI - Acquired stuttering secondary to callosal infarction. PMID- 15781842 TI - C2 myelitis presenting with neuralgiform occipital pain. PMID- 15781843 TI - Visual field defect and intracerebral hemorrhage associated with use of vardenafil (Levitra). PMID- 15781844 TI - West Nile virus presenting as opsoclonus-myoclonus cerebellar ataxia. PMID- 15781845 TI - Extreme spindles: a distinctive EEG pattern in Mycoplasma pneumoniae encephalitis. PMID- 15781846 TI - Personal history: enhancing lesions. PMID- 15781847 TI - The bioavailability of IV methylprednisolone and oral prednisone in multiple sclerosis. PMID- 15781848 TI - Cannabis for dyskinesia in Parkinson disease: a randomized double-blind crossover study. PMID- 15781849 TI - Functional imaging in PNH caused by a new FilaminA mutation. PMID- 15781850 TI - Hemorrhagic infarction in white matter following acute carbon monoxide poisoning. PMID- 15781851 TI - Most cases of dementia with hippocampal sclerosis may represent frontotemporal dementia. PMID- 15781852 TI - Proteins of the S100 family regulate the oligomerization of p53 tumor suppressor. AB - S100B protein is elevated in the brains of patients with early stages of Alzheimer's disease and Down's syndrome. S100A4 is correlated with the development of metastasis. Both proteins bind to p53 tumor suppressor. We found that both S100B and S100A4 bind to the tetramerization domain of p53 (residues 325-355) only when exposed in lower oligomerization states and so they disrupt the tetramerization of p53. In addition, S100B binds to the negative regulatory and nuclear localization domains, which results in a very tight binding to p53 protein sequences that exposed the tetramerization domain in their C terminus. Because the trafficking of p53 depends on its oligomerization state, we suggest that S100B and S100A4 could regulate the subcellular localization of p53. But, the differences in the way these proteins bind to p53 could result in S100B and S1004 having different effects on p53 function in cell-cycle control. PMID- 15781853 TI - Sumoylation: a new wrestler in the DNA repair ring. PMID- 15781854 TI - Mammalian motor neurons corelease glutamate and acetylcholine at central synapses. AB - Motor neurons (MNs) are the principal neurons in the mammalian spinal cord whose activities cause muscles to contract. In addition to their peripheral axons, MNs have central collaterals that contact inhibitory Renshaw cells and other MNs. Since its original discovery >60 years ago, it has been a general notion that acetylcholine is the only transmitter released from MN synapses both peripherally and centrally. Here, we show, using a multidisciplinary approach, that mammalian spinal MNs, in addition to acetylcholine, corelease glutamate to excite Renshaw cells and other MNs but not to excite muscles. Our study demonstrates that glutamate can be released as a functional neurotransmitter from mammalian MNs. PMID- 15781855 TI - Biography of George E. Andrews. PMID- 15781856 TI - Temperature-sensitive protein-DNA dimerizers. AB - Programmable DNA-binding polyamides coupled to short peptides have led to the creation of synthetic artificial transcription factors. A hairpin polyamide-YPWM tetrapeptide conjugate facilitates the binding of a natural transcription factor Exd to an adjacent DNA site. Such small molecules function as protein-DNA dimerizers that stabilize complexes at composite DNA binding sites. Here we investigate the role of the linker that connects the polyamide to the peptide. We find that a substantial degree of variability in the linker length is tolerated at lower temperatures. At physiological temperatures, the longest linker tested confers a "switch"-like property on the protein-DNA dimerizer, in that it abolishes the ability of the YPWM moiety to recruit the natural transcription factor to DNA. These observations provide design principles for future artificial transcription factors that can be externally regulated and can function in concert with the cellular regulatory circuitry. PMID- 15781857 TI - Quantification of cyclic and linear flows in plants. AB - A method was developed to quantify the fraction of photosystem I (PSI) centers that operate according to the cyclic or linear mode, respectively. P(700) and plastocyanin oxidation were analyzed under a weak far-red excitation (approximately eight photons per s(-1) per PSI) that induces P(700) oxidation in approximately 20 s and approximately 3 s in dark-adapted and preilluminated leaves, respectively. This finding implies that, in dark-adapted leaves, most of the electrons formed on the stromal side of PSI are transferred back to PSI through an efficient cyclic chain, whereas in preilluminated leaves, electrons are transferred to NADP and then to the Benson-Calvin cycle. Preillumination thus induces a transition from the cyclic to the linear mode. A reverse transition occurs in the dark in a time that increases with the duration and intensity of preillumination. After a approximately 10-min illumination under strong light that activates the Benson-Calvin cycle, the transition from the linear to the cyclic mode is completed in >1 h (t(1/2) approximately 30 min). The fraction of PSI involved in the cyclic process in dark-adapted leaves can be close to 100%. An apparent equilibrium constant of approximately 4 between P(700) and plastocyanin was measured during the course of the far-red illumination. This value is much lower than that computed from the midpoint redox potential of the two carriers (approximately 30). These results are interpreted assuming that chloroplasts include isolated compartments defined on the basis of the structural organization of the photosynthetic chain proposed by Albertsson [Albertsson, P. A. (2001) Trends Plant Sci. 6, 349-354]. PMID- 15781858 TI - A central role of Arabidopsis thaliana ovate family proteins in networking and subcellular localization of 3-aa loop extension homeodomain proteins. AB - The organization of living cells is based on networks of interacting molecules. Systematic analysis of protein interactions of 3-aa loop extension (TALE) homeodomain proteins, fundamental regulators of plant meristem function and leaf development, revealed a highly connected, complex network. The network includes nine members of Arabidopsis thaliana ovate family proteins (AtOFPs), a plant specific protein family, indicating a close functional connection to TALE homeodomain proteins. Evidence is provided that AtOFP1 is an essential pleiotropic developmental regulator. AtOFP1 and AtOFP5 are shown to associate with the cytoskeleton and to regulate subcellular localization of TALE homeodomain proteins, suggesting a previously unrecognized control mechanism in plant development. PMID- 15781860 TI - Sigma-pi molecular dielectric multilayers for low-voltage organic thin-film transistors. AB - Very thin (2.3-5.5 nm) self-assembled organic dielectric multilayers have been integrated into organic thin-film transistor structures to achieve sub-1-V operating characteristics. These new dielectrics are fabricated by means of layer by-layer solution phase deposition of molecular silicon precursors, resulting in smooth, nanostructurally well defined, strongly adherent, thermally stable, virtually pinhole-free, organosiloxane thin films having exceptionally large electrical capacitances (up to approximately 2,500 nF.cm(-2)), excellent insulating properties (leakage current densities as low as 10(-9) A.cm(-2)), and single-layer dielectric constant (k)of approximately 16. These 3D self-assembled multilayers enable organic thin-film transistor function at very low source drain, gate, and threshold voltages (<1 V) and are compatible with a broad variety of vapor- or solution-deposited p- and n-channel organic semiconductors. PMID- 15781859 TI - Catching RNA polymerase in the act of binding: intermediates in transcription illuminated by synchrotron footprinting. PMID- 15781861 TI - Functional complementation of Trypanosoma brucei RNA in vitro editing with recombinant RNA ligase. AB - The approximately 20S RNA ligase-containing complex (L-complex) in trypanosomatid mitochondria interacts by means of RNA linkers with at least two other multiprotein complexes to mediate the editing of mitochondrial cryptogene transcripts. The L-complex contains approximately 16 proteins, including the two RNA-editing ligases (RELs), REL1 and REL2. Leishmania tarentolae REL1 and REL2 and Trypanosoma brucei REL1 were expressed as enzymatically active tandem affinity purification-tagged proteins in a Baculovirus system. When these proteins were added to mitochondrial lysates from T. brucei procyclic cells that were depleted of the cognate endogenous ligase by RNA interference down regulation of expression, the added proteins were integrated into the L-complex, and, in the case of REL1, there was a complementation of in vitro-precleaved U insertion and U-deletion editing activities of the 20S L-complex. Integration of the recombinant proteins did not occur or occurred at a very low level with noncognate ligase-depleted L-complex or with wild-type L-complex. A C-terminal region of the T. brucei recombinant REL1 downstream of the catalytic domain was identified as being involved in integration into the L-complex. The ability to perform functional complementation in vitro provides a powerful tool for molecular dissection of the editing reaction. PMID- 15781862 TI - A low-molecular-weight compound discovered through virtual database screening inhibits Stat3 function in breast cancer cells. AB - This study focused on the screening of small-molecule inhibitors that target signal transducers and activators of transcription 3 (Stat3) in human breast carcinoma. The constitutive activation of Stat3 is frequently detected in human breast cancer cell lines as well as clinical breast cancer specimens and may play an important role in the oncogenesis of breast carcinoma. Activated Stat3 may participate in oncogenesis by stimulating cell proliferation, promoting tumor angiogenesis, and resisting apoptosis. Because a variety of human cancers are associated with constitutively active Stat3, Stat3 represents an attractive target for cancer therapy. In this study, of the nearly 429,000 compounds screened by virtual database screening, chemical samples of top 100 compounds identified as candidate small-molecule inhibitors of Stat3 were evaluated by using Stat3-dependent luciferase reporter as well as other cell-based assays. Through serial functional evaluation based on our established cell-based assays, one compound, termed STA-21, was identified as the best match for our selection criteria. Further investigation demonstrated that STA-21 inhibits Stat3 DNA binding activity, Stat3 dimerization, and Stat3-dependent luciferase activity. Moreover, STA-21 reduces the survival of breast carcinoma cells with constitutive Stat3 signaling but has minimal effect on the cells in which constitutive Stat3 signaling is absent. Together, these results demonstrate that STA-21 inhibits breast cancer cells that express constitutively active Stat3. PMID- 15781864 TI - Molecular crowding enhances native state stability and refolding rates of globular proteins. AB - The presence of macromolecules in cells geometrically restricts the available space for poplypeptide chains. To study the effects of macromolecular crowding on folding thermodynamics and kinetics, we used an off-lattice model of the all-beta sheet WW domain in the presence of large spherical particles whose interaction with the polypeptide chain is purely repulsive. At all volume fractions, phi(c), of the crowding agents the stability of the native state is enhanced. Remarkably, the refolding rates, which are larger than the value at phi(c) = 0, increase nonmonotonically as phi(c) increases, reaching a maximum at phi(c)=phi(c)(*). At high values of phi(c), the depletion-induced intramolecular attraction produces compact structures with considerable structure in the denatured state. Changes in native state stability and folding kinetics at phi(c) can be quantitatively mapped onto confinement in a volume-fraction-dependent spherical pore with radius R(s) approximately (4pi/3phi(c))(1/3) R(c) (R(c) is the radius of the crowding particles) as long as phi(c)< or =phi(c)(*). We show that the extent of native state stabilization at finite phi(c) is comparable with that in a spherical pore. In both situations, rate enhancement is due to destabilization of the denatured states with respect to phi(c) = 0. PMID- 15781863 TI - Application of genome-wide expression analysis to human health and disease. AB - The application of genome-wide expression analysis to a large-scale, multicentered program in critically ill patients poses a number of theoretical and technical challenges. We describe here an analytical and organizational approach to a systematic evaluation of the variance associated with genome-wide expression analysis specifically tailored to study human disease. We analyzed sources of variance in genome-wide expression analyses performed with commercial oligonucleotide arrays. In addition, variance in gene expression in human blood leukocytes caused by repeated sampling in the same subject, among different healthy subjects, among different leukocyte subpopulations, and the effect of traumatic injury, were also explored. We report that analytical variance caused by sample processing was acceptably small. Blood leukocyte gene expression in the same individual over a 24-h period was remarkably constant. In contrast, genome wide expression varied significantly among different subjects and leukocyte subpopulations. Expectedly, traumatic injury induced dramatic changes in apparent gene expression that were greater in magnitude than the analytical noise and interindividual variance. We demonstrate that the development of a nation-wide program for gene expression analysis with careful attention to analytical details can reduce the variance in the clinical setting to a level where patterns of gene expression are informative among different healthy human subjects, and can be studied with confidence in human disease. PMID- 15781865 TI - Identification of the mismatch repair genes PMS2 and MLH1 as p53 target genes by using serial analysis of binding elements. AB - The ability to determine the global location of transcription factor binding sites in vivo is important for a comprehensive understanding of gene regulation in human cells. We have developed a technology, called serial analysis of binding elements (SABE), involving subtractive hybridization of chromatin immunoprecipitation-enriched DNA fragments followed by the generation and analysis of concatamerized sequence tags. We applied the SABE technology to search for p53 target genes in the human genome, and have identified several previously described p53 targets in addition to numerous potentially novel targets, including the DNA mismatch repair genes MLH1 and PMS2. Both of these genes were determined to be responsive to DNA damage and p53 activation in normal human fibroblasts, and have p53-response elements within their first intron. These two genes may serve as a sensor in DNA repair mechanisms and a critical determinant for the decision between cell-cycle arrest and apoptosis. These results also demonstrate the potential for use of SABE as a broadly applicable means to globally identify regulatory elements for human transcription factors in vivo. PMID- 15781866 TI - Enhanced T cell-mediated protection against malaria in human challenges by using the recombinant poxviruses FP9 and modified vaccinia virus Ankara. AB - Malaria is a major global health problem for which an effective vaccine is required urgently. Prime-boost vaccination regimes involving plasmid DNA and recombinant modified vaccinia virus Ankara-encoding liver-stage malaria antigens have been shown to be powerfully immunogenic for T cells and capable of inducing partial protection against experimental malaria challenge in humans, manifested as a delay in time to patent parasitemia. Here, we report that substitution of plasmid DNA as the priming vector with a specific attenuated recombinant fowlpox virus, FP9, vaccine in such prime-boost regimes can elicit complete sterile protection that can last for 20 months. Protection at 20 months was associated with persisting memory but not effector T cell responses. The protective efficacy of various immunization regimes correlated with the magnitude of induced immune responses, supporting the strategy of maximizing durable T cell immunogenicity to develop more effective liver-stage vaccines against Plasmodium falciparum malaria. PMID- 15781867 TI - Mapping the interaction surface of a membrane protein: unveiling the conformational switch of phospholamban in calcium pump regulation. AB - We have used magnetic resonance to map the interaction surface of an integral membrane protein for its regulatory target, an integral membrane enzyme. Phospholamban (PLN) regulates cardiac contractility via its modulation of sarco(endo)plasmic reticulum calcium ATPase (SERCA) activity. Impairment of this regulatory process causes heart failure. To map the molecular details of the PLN/SERCA interaction, we have functionally reconstituted SERCA with labeled PLN in dodecylphosphocholine micelles for high-resolution NMR spectroscopy and in both micelles and lipid bilayers for EPR spectroscopy. Differential perturbations in NMR linewidths and chemical shifts, measured as a function of position in the PLN sequence, provide a vivid picture of extensive SERCA contacts in both cytoplasmic and transmembrane domains of PLN and provide structural insight into previously reported functional mutagenesis data. NMR and EPR data show clear and complementary evidence for a dynamic (micros-to-ms) equilibrium between two conformational states in the cytoplasmic domain of PLN. These results support the hypothesis that SERCA attracts the cytoplasmic domain of PLN away from the lipid surface, shifting the preexisting equilibrium of PLN conformers toward a structure that is poised to interact with the regulatory target. EPR shows that this conformational switch behaves similarly in micelles and lipid membranes. Based on structural and dynamics data, we propose a model in which PLN undergoes allosteric activation upon encountering SERCA. PMID- 15781868 TI - Characterizing wildfire regimes in the United States. AB - Wildfires statistics for the conterminous United States (U.S.) are examined in a spatially and temporally explicit manner. We use a high-resolution data set consisting of 88,916 U.S. Department of Agriculture Forest Service wildfires over the time period 1970-2000 and consider wildfire occurrence as a function of ecoregion (land units classified by climate, vegetation, and topography), ignition source (anthropogenic vs. lightning), and decade. For the conterminous U.S., we (i) find that wildfires exhibit robust frequency-area power-law behavior in 18 different ecoregions; (ii) use normalized power-law exponents to compare the scaling of wildfire-burned areas between ecoregions, finding a systematic change from east to west; (iii) find that wildfires in the eastern third of the U.S. have higher power-law exponents for anthropogenic vs. lightning ignition sources; and (iv) calculate recurrence intervals for wildfires of a given burned area or larger for each ecoregion, allowing for the classification of wildfire regimes for probabilistic hazard estimation in the same vein as is now used for earthquakes. PMID- 15781869 TI - Pathogen effector protein screening in yeast identifies Legionella factors that interfere with membrane trafficking. AB - Legionella pneumophila invades and replicates intracellularly in human and protozoan hosts. The bacteria use the Icm/Dot type IVB secretion system to translocate effectors that inhibit phagosome maturation and modulate host vesicle trafficking pathways. To understand how L. pneumophila modulates organelle trafficking in host cells, we carried out pathogen effector protein screening in yeast, identifying L. pneumophila genes that produced membrane trafficking [vacuole protein sorting (VPS)] defects in yeast. We identified four L. pneumophila DNA fragments that perturb sorting of vacuolar proteins. Three encode ORFs of unknown function that are translocated via the Icm/Dot transporter from Legionella into macrophages. VPS inhibitor protein (Vip) A is a coiled-coil protein, VipD is a patatin domain-containing protein, and VipF contains an acetyltransferase domain. Processing studies in yeast indicate that VipA, VipD, and VipF inhibit lysosomal protein trafficking by different mechanisms; overexpressing VipA has an effect on carboxypeptidase Y trafficking, whereas VipD interferes with multivesicular body formation at the late endosome and endoplasmic reticulum-to-Golgi body transport. Such differences highlight the multiple strategies L. pneumophila effectors use to subvert host trafficking processes. Using yeast as an effector gene discovery tool allows for a powerful, genetic approach to both the identification of virulence factors and the study of their function. PMID- 15781870 TI - CD4+ T cells mediate antibody-independent acquired immunity to pneumococcal colonization. AB - Acquired immunity to Streptococcus pneumoniae (pneumococcus) has long been assumed to depend on the presence of anticapsular antibodies. We found, however, that colonization with live pneumococci of serotypes 6B, 7F, or 14 protected mice against recolonization by any of the serotypes and that protection from acquisition of a heterologous or homologous strain did not depend on anticapsular antibody. Further, intranasal immunization by live pneumococcal colonization or by a killed, nonencapsulated whole-cell vaccine protected antibody-deficient mice against colonization, suggesting independence of antibodies to any pneumococcal antigens. Protection by intranasal immunization with whole-cell vaccine was completely abrogated in T cell-deficient mice, and in mice that were congenitally deficient in CD4(+) T cells or depleted of these cells at the time of challenge. In contrast, mice congenitally deficient in, or depleted of, CD8(+) T cells were fully protected. Protection in this model was observed beyond 2 months after immunization, arguing against innate or nonspecific immune mechanisms. Thus, we find that immunity to pneumococcal colonization can be induced in the absence of antibody, independent of the capsular type, and this protection requires the presence of CD4(+) T cells at the time of challenge. PMID- 15781871 TI - Night blindness and abnormal cone electroretinogram ON responses in patients with mutations in the GRM6 gene encoding mGluR6. AB - We report three unrelated patients with mutations in the GRM6 gene that normally encodes the glutamate receptor mGluR6. This neurotransmitter receptor has been shown previously to be present only in the synapses of the ON bipolar cell dendrites, and it mediates synaptic transmission from rod and cone photoreceptors to this type of second-order neuron. Despite the synaptic defect, best visual acuities were normal or only moderately reduced (20/15 to 20/40). The patients were night blind from an early age, and when maximally dark-adapted, they could perceive lights only with an intensity equal to or slightly dimmer than that normally detected by the cone system (i.e., 2-3 log units above normal). Electroretinograms (ERGs) in response to single brief flashes of light had clearly detectable a-waves, which are derived from photoreceptors, and greatly reduced b-waves, which are derived from the second-order inner retinal neurons. ERGs in response to sawtooth flickering light indicated a markedly reduced ON response and a nearly normal OFF response. There was no subjective delay in the perception of suddenly appearing white vs. black objects on a gray background. These patients exemplify a previously unrecognized, autosomal recessive form of congenital night blindness associated with a negative ERG waveform. PMID- 15781872 TI - Regulation of inducible nitric oxide synthase by aggresome formation. AB - Misfolding and aggregation of proteins play an important part in the pathogenesis of several genetic and degenerative diseases. Recent evidence suggests that cells have evolved a pathway that involves sequestration of aggregated proteins into specialized "holding stations" called aggresomes. Here we show that cells regulate inducible NO synthase (iNOS), an important host defense protein, through aggresome formation. iNOS aggresome formation depends on a functional dynein motor and the integrity of the microtubules. The iNOS aggresome represents a "physiologic aggresome" and thus defines a new paradigm for cellular regulation of protein processing. This study indicates that aggresome formation in response to misfolded proteins may merely represent an acceleration of an established physiologic regulatory process for specific proteins whose regulation by aggresome formation is deemed necessary by the cell. PMID- 15781873 TI - An Arabidopsis transcription factor, AtbZIP60, regulates the endoplasmic reticulum stress response in a manner unique to plants. AB - Analysis of transcripts of 75 genes encoding putative basic leucine zipper (bZIP) transcription factors in the Arabidopsis genome identified AtbZIP60, which was induced by tunicamycin. AtbZIP60 encodes a predicted protein of 295 aa with a putative transmembrane domain near its C terminus after a bZIP domain. A truncated form of AtbZIP60 without a transmembrane domain (AtbZIP60 delta C) fused with GFP localized to the nucleus, suggesting translocation of native protein to the nucleus by release from the membrane. AtbZIP60 was also induced by DTT and azetidine-2-carboxylate, which induce the endoplasmic reticulum (ER) stress response (also called the unfolded protein response). Expression of AtbZIP60 delta C clearly activated any of three BiP and two calnexin promoters in a dual luciferase assay using protoplasts of cultured cells. The induction was considered to be through cis-elements plant-specific unfolded protein response element and ER stress-response element. Interestingly, AtbZIP60 delta C also appeared to induce the expression of AtbZIP60 through an ER stress-response element-like sequence in the promoter of AtbZIP60. These characteristics of AtbZIP60 imply a signal transduction pathway of the ER stress response unique to plants. PMID- 15781874 TI - Methods to improve the detection of mild cognitive impairment. AB - We examined whether the performance of the National Institute of Aging's Consortium to Establish a Registry for Alzheimer's Disease's 10-word list (CWL), part of the consortium's neuropsychological battery, can be improved for detecting Alzheimer's disease and related disorders early. We focused on mild cognitive impairment (MCI) and mild dementia because these stages often go undetected, and their detection is important for treatment. Using standardized diagnostic criteria combined with history, physical examination, and cognitive, laboratory, and neuroimaging studies, we staged 471 community-dwelling subjects for dementia severity by using the Clinical Dementia Rating Scale. We then used correspondence analysis (CA) to derive a weighted score for each subject from their item responses over the three immediate- and one delayed-recall trials of the CWL. These CA-weighted scores were used with logistic regression to predict each subject's probability of impairment, and receiver operating characteristic analysis was used to measure accuracy. For MCI vs. normal, accuracy was 97% [confidence interval (C.I.) 97-98%], sensitivity was 94% (C.I. 93-95%), and specificity was 89% (C.I. 88-91%). For MCI/mild dementia vs. normal, accuracy was 98% (C.I. 98-99%), sensitivity was 96% (C.I. 95-97%), and specificity was 91% (C.I. 89-93%). MCI sensitivity was 12% higher (without lowering specificity) than that obtained with the delayed-recall total score (the standard method for CWL interpretation). Optimal positive and negative predictive values were 100% and at least 96.6%. These results show that CA-weighted scores can significantly improve early detection of Alzheimer's disease and related disorders. PMID- 15781875 TI - Beneficial effects of pomegranate juice on oxidation-sensitive genes and endothelial nitric oxide synthase activity at sites of perturbed shear stress. AB - Atherosclerosis is enhanced in arterial segments exposed to disturbed flow. Perturbed shear stress increases the expression of oxidation-sensitive responsive genes (such as ELK-1 and p-JUN) in the endothelium. Evidence suggests that polyphenolic antioxidants contained in the juice derived from the pomegranate can contribute to the reduction of oxidative stress and atherogenesis. The aim of the present study was to evaluate the effects of intervention with pomegranate juice (PJ) on oxidation-sensitive genes and endothelial NO synthase (eNOS) expression induced by high shear stress in vitro and in vivo. Cultured human coronary artery endothelial cells (EC) exposed to high shear stress in vitro and hypercholesterolemic mice were used in this study. PJ concentrate reduced the activation of redox-sensitive genes (ELK-1 and p-JUN) and increased eNOS expression (which was decreased by perturbed shear stress) in cultured EC and in atherosclerosis-prone areas of hypercholesterolemic mice. Moreover, oral administration of PJ to hypercholesterolemic mice at various stages of disease reduced significantly the progression of atherosclerosis. This experimental study indicates that the proatherogenic effects induced by perturbed shear stress can be reversed by chronic administration of PJ. This approach may have implications for the prevention or treatment of atherosclerosis and its clinical manifestations. PMID- 15781876 TI - Bmpr1a and Bmpr1b have overlapping functions and are essential for chondrogenesis in vivo. AB - Previous studies have demonstrated the ability of bone morphogenetic proteins (BMPs) to promote chondrogenic differentiation in vitro. However, the in vivo role of BMP signaling during chondrogenesis has been unclear. We report here that BMP signaling is essential for multiple aspects of early chondrogenesis. Whereas mice deficient in type 1 receptors Bmpr1a or Bmpr1b in cartilage are able to form intact cartilaginous elements, double mutants develop a severe generalized chondrodysplasia. The majority of skeletal elements that form through endochondral ossification are absent, and the ones that form are rudimentary. The few cartilage condensations that form in double mutants are delayed in the prechondrocytic state and never form an organized growth plate. The reduced size of mutant condensations results from increased apoptosis and decreased proliferation. Moreover, the expression of cartilage-specific extracellular matrix proteins is severely reduced in mutant elements. We demonstrate that this defect in chondrocytic differentiation can be attributed to lack of Sox9, L-Sox5, and Sox6 expression in precartilaginous condensations in double mutants. In summary, our study demonstrates that BMPR1A and BMPR1B are functionally redundant during early chondrogenesis and that BMP signaling is required for chondrocyte proliferation, survival, and differentiation in vivo. PMID- 15781877 TI - High-dose therapy and stem-cell rescue for Ewing's family of tumors in second remission. PMID- 15781878 TI - Carcinoma of the esophagus: is biology screaming in my deaf ears? PMID- 15781879 TI - A comment on the utility of recursive partitioning. PMID- 15781880 TI - Improved detection of prostate cancer using classification and regression tree analysis. AB - PURPOSE: To build a decision tree for patients suspected of having prostate cancer using classification and regression tree (CART) analysis. PATIENTS AND METHODS: Data were uniformly collected on 1,433 referred men with a serum prostate-specific antigen (PSA) levels of < or = 10 ng/mL who underwent a prostate biopsy. Factors analyzed included demographic, laboratory, and ultrasound data (ie, hypoechoic lesions and PSA density [PSAD]). Twenty percent of the data was randomly selected and reserved for study validation. CART analysis was performed in two steps, initially using PSA and digital rectal examination (DRE) alone and subsequently using the remaining variables. RESULTS: CART analysis selected a PSA cutoff of more than 1.55 ng/mL for further work-up, regardless of DRE findings. CART then selected the following subgroups at risk for a positive biopsy: (1) PSAD more than 0.165 ng/mL/cc; (2) PSAD < or = 0.165 ng/mL/cc and a hypoechoic lesion; (3) PSAD < or = 0.165 ng/mL/cc, no hypoechoic lesions, age older than 55.5 years, and prostate volume < or = 44.0 cc; and (4) PSAD < or = 0.165 ng/mL/cc, no hypoechoic lesions, age older than 55.5 years, and 50.25 cc less than prostate volume < or = 80.8 cc. In the validation data set, specificity and sensitivity were 31.3% and 96.6%, respectively. Cancers that were missed by the CART were Gleason score 6 or less in 93.4% of cases. Receiver operator characteristic curve analysis showed that CART and logistic regression models had similar accuracy (area under the curve = 0.74 v 0.72, respectively). CONCLUSION: Application of CART analysis to the prostate biopsy decision results in a significant reduction in unnecessary biopsies while retaining a high degree of sensitivity when compared with the standard of performing a biopsy of all patients with an abnormal PSA or DRE. PMID- 15781881 TI - Survival after recurrence of Ewing's sarcoma family of tumors. AB - PURPOSE: The overall survival (OS) of patients with relapsed Ewing's sarcoma family of tumors (ESFT) is poor, and the relative benefit of high-dose therapy (HDT) is controversial. PATIENTS AND METHODS: We retrospectively identified 55 consecutive ESFT patients with adequate medical records for review, who were treated at Children's Hospital and Regional Medical Center and who developed disease recurrence between January 1, 1985 and December 31, 2002. RESULTS: The median relapse-free interval (RFI) from diagnosis to first recurrence was 17 months (range, 5 to 90 months). Most recurrences were metastatic only (39 patients) or local and metastatic (10 patients). Twenty-seven patients (49%) achieved a partial or complete response to second-line treatment, with a median duration of response of 27 months (range, 5 to 119+ months). The 5-year OS rate for all relapsed patients was 23% (95% CI, 11% to 35%). By univariate analysis, improved OS was associated with response to second-line treatment versus no response (46% v 0%, respectively; P < .0001), RFI > or = 24 months versus less than 24 months (48% v 12%, respectively; P = .0001), and no metastases at initial diagnosis versus presence of metastases (31% v 12%, respectively; P = .05). Because all 13 patients who received HDT also had responsive relapse, we performed a multivariate analysis. Reduced risk of death was associated with response to second-line therapy (relative risk, 0.14; 95% CI, 0.05 to 0.40), RFI > or = 24 months (relative risk, 0.29; 95% CI, 0.13 to 0.66), and receiving HDT (relative risk, 0.26; 95% CI, 0.08 to 0.85). CONCLUSION: HDT as consolidation therapy for relapsed ESFT seems to be associated with improved OS, even after adjusting for RFI and response to second-line treatment. PMID- 15781882 TI - Complete response to neoadjuvant chemoradiotherapy in esophageal carcinoma is associated with significantly improved survival. AB - PURPOSE: Attempts to improve survival of patients with esophageal cancer have been made using induction chemoradiotherapy (CRT) followed by surgery. A large single-center experience was reviewed to determine which treatment-related variables could predict survival and recurrence. PATIENTS AND METHODS: All patients undergoing esophagectomy between January 1994 and December 2002 were reviewed. Univariate and multivariate analyses were performed using log-rank and Cox proportional hazards models, and survival curves were estimated using the Kaplan-Meier method. RESULTS: Of 171 patients with invasive cancer, 131 (77%) underwent preoperative CRT. The average age was 60 years, and most patients were male (85%). Operations performed included Ivor-Lewis (60%), transhiatal (8%), three-hole (23%), or left thoracoabdominal (8%) esophagectomy. Perioperative mortality rate was 5%. Median overall survival (OS) of the entire group was 33 months, and the 5-year OS rate was 26%. Induction CRT was associated with a 33% 5 year survival rate compared with 11% for surgery alone (P = .43). Patients downstaged to pathologic stage 0 or I had an improved OS and disease-free survival (DFS) compared with those patients who were not downstaged (P = .022). Additionally, the ability to perform an R0 resection was a significant factor for OS and DFS (n = 130; P < .0001 and P <.0002, respectively). CONCLUSION: Response to CRT and the ability to perform an R0 resection are associated with significantly improved survival in patients with esophageal carcinoma. PMID- 15781883 TI - The mechanics of sound production in Panacanthus pallicornis (Orthoptera: Tettigoniidae: Conocephalinae): the stridulatory motor patterns. AB - To examine whether sound production in katydids relies on an escapement mechanism similar to that of crickets we investigated the functional anatomy and mechanical properties of the stridulatory apparatus in the katydid Panacanthus pallicornis. Males of this species produce sustained pulses with a sharp low frequency peak of approximately approximately 5 kHz and a broad band spectrum between 15 and 25 kHz. Simultaneous recordings of movement and sound indicate that the entire stridulatory file is used for sound production and there is nearly a 1:1 correspondence between the number of cycles in a song and the number of teeth on the file. There is an overall tendency for both the spacing of teeth to increase along the file and the velocity of wing closure to increase as the scraper traverses the file. There is considerable variation, however, in the evenness of tooth spacing and in the instantaneous velocity of wing closure during sound production. The production of sustained pulses appears to depend on resonance in the right tegmen, with the left tegmen acting primarily as a damping element. This resonance is not strongly coupled to the scraper and, unlike crickets, the timing of file-scraper interactions, and therefore the phasing of energy input to wing oscillations, is variable. Similarly, the quality of the sound spectrum varies over the course of a single stridulatory wing-stroke. Based on measurements of tooth spacing on the stridulatory file and cycle-by-cycle frequency of sound output, we predicted the velocity of wing movement that would provide consistent phasing of file-scraper interactions with respect to sound radiating wing oscillations and compared this with measurements of wing velocity. Acceleration of wing velocity during stridulation results in a closer match to the velocity required for optimal phasing during a portion of the call, and this corresponds with higher amplitudes of radiated sound and the excitation of higher order modes of vibration (evident as distinct harmonic peaks in spectrograms). Our results suggest that in katydid stridulation, the movement of the scraper along the file is not regulated by an escapement mechanism as it is in crickets. Instead, katydids that produce pure-tone songs sweep their wings over a range of velocities, within which some portion matches file tooth spacing to give optimal phasing of energy input to excite a resonance in the right tegmen. PMID- 15781884 TI - A novel diuretic hormone receptor in Drosophila: evidence for conservation of CGRP signaling. AB - The Drosophila orphan G protein-coupled receptor encoded by CG17415 is related to members of the calcitonin receptor-like receptor (CLR) family. In mammals, signaling from CLR receptors depend on accessory proteins, namely the receptor activity modifying proteins (RAMPs) and receptor component protein (RCP). We tested the possibility that this Drosophila CLR might also require accessory proteins for proper function and we report that co-expression of the mammalian or Drosophila RCP or mammalian RAMPs permitted neuropeptide diuretic hormone 31 (DH31) signaling from the CG17415 receptor. RAMP subtype expression did not alter the pharmacological profile of CG17415 activation. CG17415 antibodies revealed expression within the principal cells of Malpighian tubules, further implicating DH31 as a ligand for this receptor. Immunostaining in the brain revealed an unexpected convergence of two distinct DH signaling pathways. In both the larval and adult brain, most DH31 receptor-expressing neurons produce the neuropeptide corazonin, and also express the CRFR-related receptor CG8422, which is a receptor for the neuropeptide diuretic hormone 44 (DH44). There is extensive convergence of CRF and CGRP signaling within vertebrates and we report a striking parallel in Drosophila involving DH44 (CRF) and DH31 (CGRP). Therefore, it appears that both the molecular details as well as the functional organization of CGRP signaling have been conserved. PMID- 15781885 TI - Rhodopsin patterning in central photoreceptor cells of the blowfly Calliphora vicina: cloning and characterization of Calliphora rhodopsins Rh3, Rh5 and Rh6. AB - The ommatidia that constitute the compound eyes of flies contain eight photoreceptor cells, which are divided into two classes: the peripheral photoreceptors, R1-6, and the central photoreceptors, R7 and R8. In the fruit fly, Drosophila, R1-6 express the same rhodopsin (Rh1), whilst the R7 and R8 of a given ommatidium express either Rh3 and Rh5, or Rh4 and Rh6, respectively. We have studied whether this expression pattern of rhodopsins is conserved in the blowfly Calliphora vicina. We have cloned three novel Calliphora rhodopsins, which are homologues of Drosophila Rh3, Rh5 and Rh6, with an amino acid sequence identity of 80.7%, 60.9% and 86.1%, respectively. Immunocytochemical studies with antibodies specific for Rh3, Rh5 and Rh6 revealed that Rh3 is expressed in a subset of R7 cells, while Rh5 and Rh6 are expressed in a non-overlapping subset of R8 cells. Rh3 and Rh5 are present in most cases in the same ommatidia, which account for approximately 27% of all ommatidia, and Rh6 is found in the complementary 73%. The similarity of the rhodopsin expression pattern of Calliphora with that of Drosophila suggests that the developmental mechanism regulating the terminal differentiation of R7 and R8 cells are highly conserved between these fly species. PMID- 15781886 TI - Flow patterns generated by oblate medusan jellyfish: field measurements and laboratory analyses. AB - Flow patterns generated by medusan swimmers such as jellyfish are known to differ according the morphology of the various animal species. Oblate medusae have been previously observed to generate vortex ring structures during the propulsive cycle. Owing to the inherent physical coupling between locomotor and feeding structures in these animals, the dynamics of vortex ring formation must be robustly tuned to facilitate effective functioning of both systems. To understand how this is achieved, we employed dye visualization techniques on scyphomedusae (Aurelia aurita) observed swimming in their natural marine habitat. The flow created during each propulsive cycle consists of a toroidal starting vortex formed during the power swimming stroke, followed by a stopping vortex of opposite rotational sense generated during the recovery stroke. These two vortices merge in a laterally oriented vortex superstructure that induces flow both toward the subumbrellar feeding surfaces and downstream. The lateral vortex motif discovered here appears to be critical to the dual function of the medusa bell as a flow source for feeding and propulsion. Furthermore, vortices in the animal wake have a greater volume and closer spacing than predicted by prevailing models of medusan swimming. These effects are shown to be advantageous for feeding and swimming performance, and are an important consequence of vortex interactions that have been previously neglected. PMID- 15781887 TI - Statistical analysis of modified complete randomized designs: applications to chemo-orientation studies. AB - Often experimental scientists employ a Randomized Complete Block Design (RCBD) to study the effect of treatments on different subjects. Under a 'complete randomization', the order of the apparatus setups within each block, including all replications of each treatment across all subjects, is completely randomized. However, in many experimental settings complete randomization is impractical due to the cost involved in re-adjusting the device to administer a new treatment. One typically resorts to a type of 'restricted randomization', in which multiple subjects are tested under each treatment before the apparatus is re-adjusted. The order of the treatments as well as the assignment of subjects to each block are random. If the data obtained under any type of restricted randomization are treated as if the data were collected under an RCBD with complete randomization within each block, then there is potential to increase the risk of false positives (Type I error). This is of concern to animal orientation studies and other areas such as chemical ecology where it is impractical to reset the experimental device for each subject tested. The goal of the research presented in this article is twofold: (1) to demonstrate the consequences of constructing an F-statistic based on a mean square error for testing the significance of treatment effects under the restricted randomization; (2) to describe an alternative method, based on split-plot analysis of variance, to analyze designed experiments that yield better power under the restricted randomization. The statistical analyses of simulated experiments and data involving virgin male Periplaneta americana substantiate the benefits of the alternative approach under the restricted randomization. The methodology and analysis employed for the simulated experiment is equally applicable to any organism or artificial agent tested under a restricted randomization protocol. PMID- 15781888 TI - Effect of dietary fatty acid composition on depot fat and exercise performance in a migrating songbird, the red-eyed vireo. AB - Most migrating birds accumulate lipid stores as their primary source of energy for fueling long distance flights. Lipid stores of birds during migration are composed of mostly unsaturated fatty acids; whether such a fatty acid composition enhances exercise performance of birds is unknown. We tested this hypothesis by measuring metabolic rate at rest and during intense exercise in two groups of red eyed vireos, a long-distance migratory passerine, fed either a diet containing 82% unsaturated fat (82%U), or one containing 58% unsaturated fat (58%U). Vireos fed the 82%U diet had fat stores containing (77%) unsaturated fatty acids, whereas vireos fed the 58% U diet had fat stores containing less (66%) unsaturated fatty acids. Blood metabolites measured prior to and immediately following exercise confirmed that vireos were metabolizing endogenous fat during intense exercise. Mass-specific resting metabolic rate (RMR) was similar for vireos fed the 58%U diet (2.75+/-0.32 ml O2 g(-1)h(-1)) and for vireos fed the 82%U diet (2.30+/-0.30 ml O2 g(-1) h(-1)). However, mass-specific peak metabolic rate (MR(peak)) was 25% higher in vireos fed the 58%U diet (28.55+/-1.47 ml O2 g( 1) (h-1)) than in vireos fed the 82%U diet (21.50+/-1.76 ml O2 g(-1) h(-1)). Such whole-animal energetic effects of fatty acid composition of birds suggest that the energetic cost of migration in birds may be affected by the fatty acid composition of the diet. PMID- 15781889 TI - Androgen control of immunocompetence in the male house finch, Carpodacus mexicanus Muller. AB - The immunocompetence handicap (ICH) hypothesis predicts that elevated levels of the gonadal androgen testosterone (T) entail obligatory costs, such as immunosuppression, but evidence supporting this immunosuppressive influence is equivocal. To investigate this question, adult males house finches, Carpodacus mexicanus, were exposed to short days and chronically treated with T-filled (T males; N=10) or empty (C males; N=10) Silastic capsules. Testosterone administration increased plasma T levels and the size of the cloacal protuberance, an androgen-dependent secondary sexual characteristic. To study humoral immunity, finches received injections of sheep red blood cells (SRBC) and we measured circulating concentrations of antibodies to these cells with a hemagglutination test. All males produced antibodies following four SRBC injections at weekly intervals. Antibody titers in T and C males did not differ 5 days after the fourth injection, but were 59% lower in T than C males 2 weeks later. To study cell-mediated immunity, we measured the local inflammatory response to an injection of phytohemaglutinin (PHA). This response in T and C males was similar 1 day after PHA injection, but was 58% less in T than C males 2 days following the injection. Thus, T and C males mounted similar humoral and cell-mediated immune responses, but T treatment compromised maintenance of these responses. The results, demonstrating immunosuppressive effects of elevated T, are consistent with the ICH hypothesis. PMID- 15781890 TI - Oxygen and water flux across eggshells of Manduca sexta. AB - Insect eggs must obtain oxygen across the eggshell to support embryonic development. Because eggs are small, obtaining enough oxygen would seem trivial. Recent work, however, has shown that eggs of a moth, Manduca sexta, are oxygen limited at high but realistic temperatures (32-37 degrees C) and that P(O2) drops steeply across the eggshell. Here we use theoretical and experimental approaches to partition the total resistance to oxygen flux among several steps in the oxygen cascade from environment to embryo. Standard mass-transfer analysis suggests that boundary layers of air around eggs, and around substrates to which they are attached, offer negligible resistance. Likewise, a mathematical model, parameterized using published and newly obtained morphological data, predicts that air-filled parts of the chorion also do not resist oxygen flux. This prediction was confirmed by experiments that measured rates of carbon dioxide emission from batches of eggs subjected simultaneously to hypoxia and inert gas substitution: depression of metabolic rate by hypoxia was not rescued when the diffusion coefficient of oxygen in air was doubled by substituting helium for nitrogen. The model did predict, however, that a set of subchoral layers (a crystalline chorionic layer, a wax layer and the vitelline membrane) could account for most or all of the total resistance to oxygen flux. Support for this prediction was obtained from two sequential experiments. First, eggs extracted with chloroform:methanol had highly elevated rates of water loss, suggesting that indeed eggs of M. sexta are waterproofed by wax. Second, rates of water loss and carbon dioxide emission from batches of eggs, measured from laying to hatching, changed in parallel over development. These data suggest that a single layer, likely a wax layer or a combination of wax and other subchoral layers, provides the main resistance to water efflux and oxygen influx. PMID- 15781891 TI - Testing the hindlimb-strength hypothesis: non-aerial locomotion by Chiroptera is not constrained by the dimensions of the femur or tibia. AB - In the evolution of flight bats appear to have suffered a trade-off; they have become poor crawlers relative to terrestrial mammals. Capable walking does occur in a few disparate taxa, including the vampire bats, but the vast majority of bats are able only to shuffle awkwardly along the ground, and the morphological bases of differences in crawling ability are not currently understood. One widely cited hypothesis suggests that the femora of most bats are too weak to withstand the compressive forces that occur during terrestrial locomotion, and that the vampire bats can walk because they possess more robust hindlimb skeletons. We tested a prediction of the hindlimb-strength hypothesis: that during locomotion, the forces produced by the hindlimbs of vampire bats should be larger than those produced by the legs of poorly crawling bats. Using force plates we compared the hindlimb forces produced by two species of vampire bats that walk well, Desmodus rotundus (N=8) and Diaemus youngi (N=2), to the hindlimb forces produced during over-ground shuffling by a similarly sized bat that is a poor walker (Pteronotus parnellii; N=6). Peak hindlimb forces produced by P. parnellii were larger (ANOVA; P<0.05; N=65) and more variable (93.5+/-36.6% body weight, mean +/- s.d.) than those of D. rotundus (69.3+/-8.1%) or D. youngi (75.0+/-6.2%). Interestingly, the vertical components of peak force were equivalent among species (P>0.6), indicating similar roles for support of body weight by the hindlimbs in the three species. We also used a simple engineering model of bending stress to evaluate the support capabilities of the hindlimb skeleton from the dimensions of 113 museum specimens in 50 species. We found that the hindlimb bones of vampires are not built to withstand larger forces than those of species that crawl poorly. Our results show that the legs of poorly crawling bats should be able to withstand the forces produced during coordinated crawling of the type used by the agile vampires, and this indicates that some mechanism other than hindlimb bone thickness, such as myology of the pectoral girdle, limits the ability of most bats to crawl. PMID- 15781892 TI - Echolocation call intensity in the aerial hawking bat Eptesicus bottae (Vespertilionidae) studied using stereo videogrammetry. AB - Aerial hawking bats use intense echolocation calls to search for insect prey. Their calls have evolved into the most intense airborne animal vocalisations. Yet our knowledge about call intensities in the field is restricted to a small number of species. We describe a novel stereo videogrammetry method used to study flight and echolocation behaviour, and to measure call source levels of the aerial hawking bat Eptesicus bottae (Vespertilionidae). Bats flew close to their predicted minimum power speed. Source level increased with call duration; the loudest call of E. bottae was at 133 dB peSPL. The calculated maximum detection distance for large flying objects (e.g. large prey, conspecifics) was up to 21 m. The corresponding maximum echo delay is almost exactly the duration of one wing beat in E. bottae and this also is its preferred pulse interval. These results, obtained by using videogrammetry to track bats in the field, corroborate earlier findings from other species from acoustic tracking methods. PMID- 15781893 TI - Eel migration to the Sargasso: remarkably high swimming efficiency and low energy costs. AB - One of the mysteries of the animal kingdom is the long-distance migration (5000 6000 km) of the European eel Anguilla anguilla L. from the coasts of Europe to its spawning grounds in the Sargasso Sea. The only evidence for the location of the spawning site of the European eel in the Sargasso Sea is the discovery by Johannes Schmidt at the beginning of the previous century of the smallest eel larvae (leptocephali) near the Sargasso Sea. For years it has been questioned whether the fasting eels have sufficient energy reserves to cover this enormous distance. We have tested Schmidt's theory by placing eels in swim tunnels in the laboratory and allowing them to make a simulated migration of 5500 km. We find that eels swim 4-6 times more efficiently than non-eel-like fish. Our findings are an important advance in this field because they remove a central objection to Schmidt's theory by showing that their energy reserves are, in principle, sufficient for the migration. Conclusive proof of the Sargasso Sea theory is likely to come from satellite tracking technology. PMID- 15781894 TI - Temporal and spatial changes in the expression pattern of multiple red and green subtype opsin genes during zebrafish development. AB - Zebrafish have two red, LWS-1 and LWS-2, and four green, RH2-1, RH2-2, RH2-3 and RH2-4, opsin genes encoding photopigments with distinct absorption spectra. Occurrence of opsin subtypes by gene duplication is characteristic of fish but little is known whether the subtypes are expressed differently in the retina, either spatially or temporally. Here we show by in situ hybridization the dynamic expression patterns of the opsin subtypes in the zebrafish retina. Expression of red type opsins is initiated with the shorter-wavelength subtype LWS-2, followed by the longer-wavelength subtype LWS-1. In the adult retina, LWS-2 was expressed in the central to dorsal area and LWS-1 in the ventral and peripheral areas. Expression patterns of green type opsins were similar to those of the red type opsins. The expression started with the shortest wavelength subtype RH2-1 followed by the longer wavelength ones, and in the adult retina, the shorter wavelength subtypes (RH2-1 and RH2-2) were expressed in the central to dorsal area and longer wavelength subtypes (RH2-3 and RH2-4) in the ventral and peripheral areas. These results provide the framework for subsequent studies of opsin gene regulation and for probing functional rationale of the developmental changes by using the power of zebrafish genetics. PMID- 15781895 TI - Central pattern generator for swimming in Melibe. AB - The nudibranch mollusc Melibe leonina swims by bending from side to side. We have identified a network of neurons that appears to constitute the central pattern generator (CPG) for this locomotor behavior, one of only a few such networks to be described in cellular detail. The network consists of two pairs of interneurons, termed 'swim interneuron 1' (sint1) and 'swim interneuron 2' (sint2), arranged around a plane of bilateral symmetry. Interneurons on one side of the brain, which includes the paired cerebral, pleural and pedal ganglia, coordinate bending movements toward the same side and communicate via non rectifying electrical synapses. Interneurons on opposite sides of the brain coordinate antagonistic movements and communicate over mutually inhibitory synaptic pathways. Several criteria were used to identify members of the swim CPG, the most important being the ability to shift the phase of swimming behavior in a quantitative fashion by briefly altering the firing pattern of an individual neuron. Strong depolarization of any of the interneurons produces an ipsilateral swimming movement during which the several components of the motor act occur in sequence. Strong hyperpolarization causes swimming to stop and leaves the animal contracted to the opposite side for the duration of the hyperpolarization. The four swim interneurons make appropriate synaptic connections with motoneurons, exciting synergists and inhibiting antagonists. Finally, these are the only neurons that were found to have this set of properties in spite of concerted efforts to sample widely in the Melibe CNS. This led us to conclude that these four cells constitute the CPG for swimming. While sint1 and sint2 work together during swimming, they play different roles in the generation of other behaviors. Sint1 is normally silent when the animal is crawling on a surface but it depolarizes and begins to fire in strong bursts once the foot is dislodged and the animal begins to swim. Sint2 also fires in bursts during swimming, but it is not silent in non-swimming animals. Instead activity in sint2 is correlated with turning movements as the animal crawls on a surface. This suggests that the Melibe motor system is organized in a hierarchy and that the alternating movements characteristic of swimming emerge when activity in sint1 and sint2 is bound together. PMID- 15781896 TI - Development of the acoustically evoked behavioral response in zebrafish to pure tones. AB - Zebrafish (Danio rerio) were placed in small wells that could be driven vertically with a series of calibrated sinusoids. Video images of the fish were obtained and analyzed to determine the levels and frequencies at which the fish responded to the stimulus tones. It was found that fish 4 days post fertilization (dpf) did not respond to the stimulus tones, whereas fish 5 dpf to adult did respond. It was further found that the stimulus thresholds and frequency bandwidth to which the fish responded did not change from 5 dpf to adult; indicating that the otolithic organ adaptations for high-frequency hearing are already present in larval fish. Deflating the swimbladders in adult fish eliminated their response, which is consistent with sensing sound pressure. Deflating the swimbladder in larval fish did not affect their thresholds, which is consistent with sensing the particle motion of the fluid directly. Because adult fish with Weberian ossicles have a greater input to the inner ear for a given sound pressure level (SPL), the finding that the adult and larval fish respond at the same SPL with intact swimbladders suggests that the acoustic startle response threshold is adjusted as the fish develop in order to maintain appropriate reactions to relevant stimuli. PMID- 15781897 TI - Cardiorespiratory and metabolic reactions during entrance into torpor in dormice, Glis glis. AB - Dormice voluntarily enter torpor at ambient temperatures ranging between 0-28 degrees C. This study describes heart rate, ventilation frequency, O2-consumption (defined as metabolic rate), CO2-production and body temperature during entrance into torpor. Their temporal relationship was analysed during the time course of metabolic depression at different ambient temperatures. Body temperature and heart rate were measured in unrestrained dormice with implanted transmitter. Ventilation frequency was monitored by total body plethysmography or infrared video monitoring. To compare entries into torpor at different T(a) these periods were distinguished into four different phases: the resting phase prior to torpor, the phase of pre-torpor adjustments, the reduction phase and the phase of steady state torpor. In the pre-torpor phase, dormice increased their ventilation, metabolic rate and heart rate, indicating that the torpid state is initiated by an enhanced metabolic activity for about an hour. This was followed by a rapid reduction of ventilation, metabolism and heart rate, which reached their minimum values long before body temperature completed its decline. The results of the present study show that the entrance into torpor is caused by an active respiratory, cardiac and metabolic depression. PMID- 15781898 TI - Respiratory changes throughout ontogeny in the tobacco hornworm caterpillar, Manduca sexta. AB - The respiratory system of growing caterpillars is challenged in two distinct ways as they develop from hatchlings to fifth instars preparing for pupation. First, across instars, body sizes and tracheal lengths increase substantially. Second, within each instar, animal mass can more than double while major tracheal respiratory system structures, such as spiracles and large tracheae, are fixed in size until molting. To test whether these growth processes result in a decrease in O2 delivery capacity relative to tissue oxygen needs, we exposed feeding Manduca sexta larvae of various ages to decreasing levels of atmospheric O2 and measured their metabolic rate and ability to feed. We found that near the beginning of all instars, M. sexta were able to maintain gas exchange and feed down to approximately 5 kPa O2, indicating that these insects are able to match tracheal O2 delivery to increased metabolic rates across instars. However, gas exchange and feeding of caterpillars nearing the molt were limited at much higher O2 levels (up to 15 kPa O2), suggesting that caterpillars have limited capacities to increase tracheal O2 delivery as O2 consumption rates increase within instars. It seems possible that the safety margin for O2 delivery may disappear completely in the last hours before ecdysis, providing an ultimate if not proximate explanation for the necessity of molting. PMID- 15781899 TI - Undetectable apolipoprotein A-I gene expression suggests an unusual mechanism of dietary lipid mobilisation in the intestine of Cyprinus carpio. AB - High density lipoprotein (HDL) has been shown to play an important role in the dietary lipid mobilisation in the carp. In spite of this, previous studies have failed to demonstrate the synthesis of the major protein component of HDL, apolipoprotein A-I (apoA-I), in the proximal intestine of the carp. Therefore, the aim of the present study was to evaluate the expression of apoA-I throughout the entire intestine. Curiously, no transcription of the apoA-I gene could be detected either by northern blot or RT-PCR assays in the intestinal mucosa, in clear contrast with the abundant cytosolic immunoreactive apoA-I detected in almost all intestinal segments, which suggests a different origin for this protein. In addition, the detection of specific, but low affinity, binding sites for apoA-I in the carp intestinal brush-border membranes (BBM), and the strong interaction with BBM, which is highly dependent on temperature, points to an important contribution of membrane lipids in apoA-I binding to the intestinal mucosa. This idea was reinforced by the ability of carp apoA-I to associate with multilamellar phospholipid vesicles. PMID- 15781901 TI - Nursing intervention research. PMID- 15781900 TI - Spiders on a treadmill: influence of running activity on metabolic rates in Pardosa lugubris (Araneae, Lycosidae) and Marpissa muscosa (Araneae, Salticidae). AB - The CO2 release of the well-tracheated jumping spider, Marpissa muscosa, and the poorly tracheated, Pardosa lugubris, was tested while animals were running on a treadmill at three different speeds and under a selective elimination of lungs or tracheae. Thus, the influence of a well-developed tracheal system on the metabolism during physical exercise was examined. The CO2 release in intact animals increased with the running speed in both species. The costs of transport (COT) running at the maximal sustainable speed were nearly twice as big in M. muscosa as in P. lugubris. Elimination of one lung by sealing resulted in reduced COT and running times, and increasing anaerobic proportions in metabolism. Effects were greater in P. lugubris than in M. muscosa, indicating that tracheae compensate partly for the lacking lung capacity. Sealing of the tracheae in M. muscosa reduced the COT and the running times only at the highest speed. Results indicate that tracheae in M. muscosa support the aerobic metabolism only at the most intense physical exercise. At low and medium activity, tracheae may play their main role in the local supply of organs that are not involved in running activity. PMID- 15781902 TI - Exercise and functional tasks among adults who are functionally limited. AB - This study compared the efficacy of 16 weeks of either resistance training, aerobic walking, or combined resistance training and aerobic walking on the performance of functional tasks among adults age 65 years and older with limited functional ability. One hundred thirty-one older adult individuals were randomized into four groups: resistance training, aerobic walking, combined resistance and aerobic walking groups, or a nonexercise control group. Each of the exercise groups documented 70% compliance with their respective exercise intervention, which included three weekly exercise sessions. At baseline, and 8 and 16 weeks following baseline, all participants completed six assessments of their functional ability. Analysis of covariance indicated that all three exercise intervention groups significantly improved measures of functional ability, with the resistance group demonstrating the most consistent gains over the six measures. These findings indicate that older adults who are functionally limited can improve their functional ability through a variety of types of exercise. PMID- 15781903 TI - Mediator analysis of Moms on the move. AB - This study examined whether improvements in physical activity discovered in a test of Moms on the Move were mediated by the behavioral constructs from which the intervention was derived. Identifying mediator variables is vital for intervention planning. The intervention was derived from the Transtheoretical Model (TTM) of behavior change and promoted moderately intense activities like walking. Sedentary mothers with children enrolled in the Women, Infants, and Children program (N = 44) were randomly assigned to the experimental intervention or attention control. Large effect sizes were reported for improvements in physical activity and changes in TTM constructs. This study examined whether the physical activity improvements were mediated by the behavioral constructs. Statistical analyses used bivariate correlation coefficients and two-stage multiple linear regression. These exploratory findings did not support the hypothesis that increased physical activity, which was associated with the experimental intervention, was mediated by the TTM constructs. PMID- 15781904 TI - Retention of minority participants in clinical research studies. AB - Recruitment of minority participants for clinical research studies has been the topic of several analytical works. Yet retention of participants, most notably minority and underserved populations, is less reported and understood, even though these populations have elevated health risks. This article describes two related, intervention-based formative research projects in which researchers used treatment theory to address issues of recruitment and retention of minority women participants in an exercise program to reduce obesity. Treatment theory incorporates a model of health promotion that allows investigators to identify and control sources of extraneous variables. The authors' research demonstrates that treatment theory can improve retention of minority women participants by considering critical inputs, mediating processes, and substantive participant characteristics in intervention design. PMID- 15781906 TI - Improving resident outcomes with GAPN organization level interventions. AB - This research tested the effectiveness of the second tier of interventions in a two-tiered nursing intervention model designed to improve quality of care for residents in long-term care facilities (LTCFs). The first tier of the model called for gerontological advanced practice nurses (GAPNs) to provide direct care and to teach staff to implement care protocols for residents with incontinence, pressure ulcers, depression, and aggression. Results of the first-tier study indicated significant improvement in resident outcomes in incontinence, pressure ulcers, and aggression. In the second tier, GAPNs added a set of organization level (OL) interventions including membership on the LTCF quality assurance committee and collaborating with staff on problem-solving teams. Analysis following the addition of OL interventions revealed significant improvement in both depression scores and in the trajectory of depression in residents of the LTCF where OL interventions were used. PMID- 15781905 TI - Claustrophobia and adherence to CPAP treatment. AB - This study evaluated the effect of claustrophobia, an abnormal dread or fear of closed spaces, on adherence to continuous positive airway pressure (CPAP) therapy. The design was a secondary analysis of data from a prospective study of participants (N = 153) that completed 3 months of CPAP therapy from seven sleep disorders centers in the United States and Canada. A 15-item subscale adapted from the Fear and Avoidance Scale measured claustrophobic tendencies pre-CPAP treatment and again after 3 months. An overt monitor attached to the CPAP machines recorded mask-on CPAP adherence. There was a statistically significant difference in claustrophobia scores by adherence group (< 2 hours, 2 to < 5 hours, > or = 5 hours) and time period (pre-CPAP and after 3 months CPAP). Poor CPAP adherence (< 2 hours per night)was more than two times higher in participants with a claustrophobia score > or = 25. Identification of persons with increased claustrophobia tendencies and targeted interventions may increase adherence. PMID- 15781907 TI - Strategies for maintaining integrity of a behavioral intervention. AB - The purpose of this article is to address issues in maintaining the integrity of a behavioral intervention. Examples are provided from a recently completed study on how to train research staff and monitor the integrity of the intervention. In this exemplar, the integrity of the behavioral intervention was addressed by the design of the study and research staff training. Throughout the study, the integrity of the behavioral intervention was monitored by delineating a checklist of topics that must be addressed, critiquing audiotapes of the intervention, and discussing incomplete or missing elements with the research staff. PMID- 15781908 TI - Selecting control interventions for clinical outcome studies. AB - In the current research environment the design and management of control groups is becoming more complex. The selection of a control group design is dependent on study goals, presence and quality of existing interventions, urgency of the problem or issue being addressed by the intervention, and factors related to the study site. The purpose of the presentation is to identify various approaches to the design of control groups in experimental studies and to identify their strengths, limitations, and applications. A case study exemplifies the issues associated with control group selection and design. PMID- 15781909 TI - Benefits and challenges in developing a program of research. AB - The ultimate goals of nursing research are knowledge generation and improvement in nursing practice. Designing studies that provide the evidence needed for practice change and that have clear implications for immediate application to current practice environments is particularly challenging. Research programs that consist of sequential studies, each building on and expanding on the results of the previous work, offer the greatest promise for generating understanding of the human phenomenon relevant to nursing practice. The authors review their experience with a series of studies of inpatient and postdischarge needs and interventions associated with chronic critical illness to illustrate the benefits of developing a longitudinal research program as well as the importance of strategies that will foster application of results. Recommendations for developing such a program are discussed. PMID- 15781910 TI - Is the prothrombin 19911A>G polymorphism a functional noncoding variant? PMID- 15781911 TI - Treatment with cyclosporin A cream for the cutaneous reactions associated with imatinib therapy. PMID- 15781913 TI - Commentary: David Kupfer, Ph.D., 1925-2004. PMID- 15781912 TI - Quantitative RT-PCR analysis of activation-induced cytidine deaminase expression in tissue samples from mantle cell lymphoma and B-cell chronic lymphocytic leukemia patients. PMID- 15781914 TI - Conflicts of care. PMID- 15781915 TI - Managing chronic pain in children: the challenge of delivering chronic care in a "modernising" healthcare system. PMID- 15781916 TI - Improving the outcome of pneumococcal meningitis. PMID- 15781917 TI - Consumerism in healthcare can be detrimental to child health: lessons from children with functional abdominal pain. AB - AIMS: To determine prognostic indicators in children with severe functional abdominal pain (FAP) and to test the hypothesis that "healthcare consumerism" in these families might be deleterious to the child. METHODS: Retrospective analysis of a cohort of 23 children aged <16 years fulfilling the Rome II diagnostic criteria for FAP during the period December 1997 to February 2001. Poor outcome was defined as continued pain and failure to return to normal functioning >12 months after onset. RESULTS: Poor outcome was associated with refusal to engage with psychological services, involvement of more than three consultants, lodging of a manipulative complaint with hospital management by the child's family, and lack of development of insight into psychosocial influences on symptoms. Three of four adverse prognostic indicators reflected healthcare consumerism by the families. CONCLUSIONS: Actions of families who lack insight into their child's illness may perpetuate FAP in childhood. A culture of parental consumerism in healthcare, however well intentioned, needs to be accompanied by robust systems to protect the interests of the child. PMID- 15781919 TI - Twenty years of a multidisciplinary paediatric diabetes home care unit. AB - As only a minority of patients with type 1 diabetes are unwell at diagnosis, these patients could be managed at home if appropriate facilities were available. A multidisciplinary diabetes home care service was established over 20 years ago at Birmingham Children's Hospital, to support children with diabetes mellitus within the home environment from diagnosis, reducing emotional upset and separation. Despite increase in the size and distribution of the unit over this time (from 230 to 400 patients (now spread over two hospitals)), the proportion of newly diagnosed children managed wholly at home (median 43%; range 31-67%), and the reduction in number and duration of admissions has been sustained (readmission rate with diabetic ketoacidosis 4.1 bed-days per 100 patients/year; range 2.9-7.1), with no deterioration in overall blood glucose control. In this way the savings achieved by reductions in expensive hospital bed occupancy have more than offset the costs of maintaining the unit. PMID- 15781918 TI - DT5aP-Hib-IPV and MCC vaccines: preterm infants' response to accelerated immunisation. AB - AIMS: To describe the immune response of preterm infants to combined diphtheria/tetanus/5 component acellular pertussis-Haemophilus influenzae type b inactivated polio vaccine (DT5aP-Hib-IPV) and meningococcal serogroup C conjugate vaccine (MCC) under accelerated schedule. To compare results with term infants immunised with DT5aP-Hib-IPV and with historical data from preterm infants immunised with a DT3 component aP-Hib vaccine. METHODS: Prospective observational study in preterm infants born at <32 weeks gestation with comparison to contemporary cohort of term infants. DT5aP-Hib-IPV and MCC vaccines were given at 2, 3, and 4 months. RESULTS: Fifty preterm infants (mean gestational age 28.5 weeks) completed the study. After three doses of vaccines Hib polysaccharide IgG geometric mean concentration (GMC) was 1.21 microg/ml with 80% > or =0.15 microg/ml; MCC serum bactericidal assay geometric mean titre (GMT) was 1245 with 100% > or =8. All infants achieved protective titres to diphtheria, tetanus, and the three poliovirus types with > or =80% achieving protective rises in IgG against the five pertussis antigens. CONCLUSION: Preterm infants immunised with DT5aP-Hib-IPV and MCC vaccines show IgG responses to Hib and MCC greater than seen historically in both term and preterm infants with a DT3aP-Hib vaccine, and for pertussis antigens and poliovirus type 1 responses similar to that seen in term infants immunised with DT5aP-Hib-IPV. Responses to poliovirus types 2 and 3 are reduced, but all infants achieved protective titres. PMID- 15781920 TI - Pre- and postoperative developmental attainment in sagittal synostosis. AB - AIMS: To investigate whether sagittal synostosis (SS) has consequences for children's mental and psychomotor development, and whether surgery has any impact on this. METHODS: The study involved 28 children with SS who underwent corrective surgery at a mean age of 8.0 (SD 7.16) months, and 28 normal controls. All the children with SS were assessed pre- and postoperatively using the Griffiths Mental Development Scales. The controls were assessed on one occasion, at an age matched with individuals in the patient group at the time of the preoperative assessment. A further control group consisted of 13 children with SS, who had received developmental assessment on two or more occasions without surgical intervention. RESULTS: The data indicated that children with SS have significantly poorer gross locomotor function than the normal controls. Following surgical intervention the deficit was shown to have resolved; consistent with this a lesser improvement in eye-hand coordination and performance skills was shown. Overall developmental attainment also improved postoperatively. The children with SS who did not receive surgery did not show any improvement in development. CONCLUSIONS: The study shows improved developmental attainment following surgical correction of SS, which may therefore be more than a cosmetic procedure. PMID- 15781921 TI - Reliability and validity of health status measurement by the TAPQOL. AB - BACKGROUND: In addition to clinical measures in the evaluation of paediatric interventions, health related quality of life (HRQoL) is an important outcome. The TAPQOL (TNO-AZL Preschool children Quality of Life) was developed to measure HRQoL in preschool children. It is a generic instrument consisting of 12 scales that cover the domains physical, social, cognitive, and emotional functioning. AIMS: To evaluate the feasibility, score distribution, internal consistency, test retest reliability, and discriminative and concurrent validity of the TAPQOL multi-item scales in preschool children, aged 2-48 months. Also to evaluate the feasibility, reliability, and validity separately for infants (2-12 months old) and toddlers (12-48 months old). METHODS: Parents of a random general population sample of 500 preschool children were sent a questionnaire by mail. A random subgroup of 159 parents who participated received a retest after two weeks. RESULTS: The response rate was 83% at the test and 75% at the retest. There were few missing answers. Six scales showed ceiling effects. Nine scales had Cronbach's alphas >0.70. In general, score distributions and Cronbach's alphas were comparable for infants and toddlers. Test-retest showed no significant differences in mean scale scores; two scales had intra-class correlations <0.50. Five scales showed significant differences between children with no conditions versus children with two or more parent reported chronic conditions. CONCLUSION: Results showed that the TAPQOL is a feasible instrument to measure HRQoL and support the reliability and discriminative validity of the majority of its scales for infants as well as toddlers. PMID- 15781923 TI - A randomised study of the impact of different styles of patient information leaflets for randomised controlled trials on children's understanding. PMID- 15781922 TI - Major differences in prevalence of overweight according to nationality in preschool children living in Germany: determinants and public health implications. AB - AIMS: To investigate the prevalence of overweight according to nationality in preschool children living in Germany, and to establish the determinants responsible for differences in body mass index. METHODS: The study was performed within the context of the 2001/2002 obligatory health examination before school entry in the city of Aachen, Germany. Of 2020 eligible children 1979 children were recruited (participation rate: 98%). Children's height and weight were measured using a standardised protocol. The parents completed a standardised questionnaire on sociodemographic factors and possible determinants of nutritional status. Being overweight was defined according to age and sex specific reference values for German children as well as according to international reference values. RESULTS: The study population included 452 (22.9%) children with other than German nationality. Among these children the prevalence of overweight was twice as high than among German children (14.8% v 7.2%). Prevalence of most known risk factors for overweight, such as low physical activity, high consumption of soft drinks, and frequent visits to fast-food restaurants was higher in the children with other nationalities than in the German children. Multivariate analyses revealed that most of the difference in prevalence of obesity by nationality is explained by known risk factors of overweight, especially education of mother and watching TV. CONCLUSIONS: The apparent ethnic differences could be explained by two non-ethnic but socioeconomic factors. In preventing overweight in children, there is the need to identify and deal with high risk environments rather than high risk ethnic groups. PMID- 15781924 TI - Prevalence of severe fatigue in primary care. AB - A postal survey of 1024 UK GP practices showed the prevalence of medically unexplained severe fatigue over three months in 5-19 year olds to be 62/100,000. Cases were predominantly adolescent girls and were more likely to come from practices in less deprived areas, which could reflect consulting behaviours. PMID- 15781925 TI - A feasibility study comparing two treatment approaches for chronic fatigue syndrome in adolescents. PMID- 15781926 TI - Toys and potentially lethal games. PMID- 15781929 TI - Bronchial artery to bronchial vein fistula secondary to lung nodule resection. PMID- 15781928 TI - Corticosteroids in the management of the paediatric epilepsies. AB - Corticosteroids (predominantly prednisolone and hydrocortisone) and adrenocorticotropic hormone (ACTH) have been used in the treatment of the epilepsies for over 50 years. Over the past 30 years most reports have focused on epilepsy syndromes and epileptic encephalopathies resistant to treatment with the more conventional anticonvulsant and antiepileptic drugs (AEDs) and specifically West syndrome. There has been relatively little attention on the role of corticosteroids in treating other epilepsies. PMID- 15781927 TI - Bone mass acquisition in healthy children. AB - Although 80% of the variance in bone mass is determined genetically, there are many other factors which influence the accumulation of bone in early life and affect future risks of osteoporosis. This review considers the genetic, fetal, and environmental influences on bone mass acquisition in healthy children, and highlights important areas where paediatricians may have a role by counselling children and their families to adopt a healthy lifestyle which promotes bone health. PMID- 15781930 TI - Managing obesity in secondary care: a personal practice. AB - Obesity is becoming a common clinical headache for child health professionals, as the increase in professional and media concern has not been matched by effective clinical solutions. Added to this is considerable confusion about both the definitions of obesity and about which groups of children and adolescents actually require treatment. Little guidance exists for paediatricians and other child health specialists on the assessment and management of child and adolescent obesity. Guidelines produced by the American Academy of Pediatrics (AAP) are useful, but require modification for non-American clinical practice. In the UK, the Royal College of Paediatrics and Child Health has recently issued excellent brief guidance on managing obesity in primary care, as has the Scottish Intercollegiate Guidelines Network. PMID- 15781931 TI - A population based study of the impact of corticosteroid therapy and delayed diagnosis on the outcome of childhood pneumococcal meningitis. AB - BACKGROUND: Despite an extensive literature, the impact of both adjunctive steroid therapy and delayed diagnosis on the outcome of childhood pneumococcal meningitis is controversial. AIM: To determine the independent contribution of corticosteroid therapy and delayed diagnosis on the outcome of childhood pneumococcal meningitis in a representative population with good access to medical services. METHODS: Data were obtained from laboratories and hospital records to assemble a population register in Sydney, Australia, 1994-99. Follow up questionnaires were completed by attending physicians. RESULTS: A total of 122 cases of pneumococcal meningitis aged 0-14 years were identified. Almost 50% of 120 children with available records either died (n = 15) or had permanent neurological impairment (n = 39). Early use (before or with parenteral antibiotics) of corticosteroids protected against death or severe morbidity (adjusted OR 0.21, 95% CI 0.05 to 0.77). Delayed diagnosis was associated with increased morbidity in survivors (OR 3.4, 95% CI 1.03 to 11.4) but not with increased mortality. CONCLUSION: In a population with good access to health care and after adjusting for other known prognostic variables, early recognition of pneumococcal meningitis and treatment with adjunctive dexamethasone significantly improves outcomes. These data add to those from randomised controlled trials. Implementation requires development of protocols and guidelines for use in emergency departments. PMID- 15781932 TI - Effect of an IMCI intervention on quality of care across four districts in Cape Town, South Africa. AB - AIMS: To measure the change in quality of care provided to sick children as a result of the routine implementation of the IMCI intervention. METHODS: Structured observations of consultations with sick children, exit interviews with caregivers, and facility reviews were conducted both before and after IMCI intervention in four health districts in Cape Town. Interventions were case management training, orientation courses for supervisors and medical officers, and some reorganisation of management systems. RESULTS: Twenty one nurses in 21 clinics were observed before and after the IMCI intervention; 90 and 70 child observations were conducted before and after IMCI intervention respectively. There was a marked improvement in assessment of danger signs in sick children (7% before versus 72% after), assessment of co-morbidity (integrated score 5.2 versus 8.2), rational prescribing (62% versus 84%), and starting treatment in the clinic (40% versus 70%). However there was no change in the treatment of anaemia or the prescribing of vitamin A or counselling of caregivers. There was no change in the knowledge of caregivers regarding medication or when to return to the health facility. Facilities were well stocked and supervision regular both before and after IMCI. CONCLUSION: This study has shown that under normal operating conditions and in a context of good facility infrastructure and management support, IMCI is associated with improvements in some important aspects of care. PMID- 15781933 TI - Thrombophilia and first arterial ischaemic stroke: a systematic review. AB - AIMS: To undertake a systematic review of the literature reporting the prevalence of thrombophilia in children with a first arterial ischaemic stroke (AIS). METHODS: Systematic review of case-control studies reporting data for prevalence of protein C, S, and antithrombin (AT) deficiencies, activated protein C resistance (APCr), total plasma homocysteine >95th centile, the thrombophilic mutations factor V1691 GA, prothrombin 20210GA, and MTHFR C677T in children with first, radiologically confirmed, AIS. RESULTS: Of 1437 potentially relevant citations, 18 met inclusion criteria. A total of 3235 patients and 9019 controls had been studied. Results of meta-analyses were expressed as pooled odds ratios (OR) relating the prevalence of the thrombophilic condition in children with AIS to that in controls. The pooled OR (and 95% CI) were: protein C deficiency, 6.49 (2.96 to 14.27); protein S deficiency, 1.14 (0.34 to 3.80); AT deficiency, 1.02 (0.28 to 3.67); APCr, 1.34 (0.16 to 11.52); FV1691 GA, 1.22 (0.80 to 1.87); PT20210GA, 1.10 (0.51 to 2.34); MTHFR C677T, 1.70 (1.23 to 2.34); and total plasma homocysteine >95th centile, 1.36 (0.53 to 3.51). There was no statistical heterogeneity within these data. CONCLUSIONS: All factors examined were more common in children with first AIS than in controls, and significantly so for protein C deficiency and the MTHFR C677T mutation. The implications of thrombophilia for prognosis and recurrence need to be established before clinical recommendations can be made regarding investigation and treatment of children with AIS. PMID- 15781935 TI - Basilar artery infarction. PMID- 15781934 TI - Observational study of suspected maltreatment in Italian paediatric emergency departments. AB - AIMS: To evaluate how often children seen in paediatric accident & emergency (A&E) departments were suspected of abuse or neglect, and to explore some of the correlates of suspected child maltreatment. METHODS: Multicentre, cross-sectional study of 15 randomised census days during a six month period. Trained research assistants working with local paediatric staff completed a purpose made anonymised checklist covering sociodemographic and medical information. A six point suspicion index was used to rate compatibility with child maltreatment based on the occurrence of observable harm. Statistical analysis was carried out on the basis that a score of 4 or more was suspicious of child maltreatment. Nineteen hospitals provided standardised paediatric A&E consultation data on 0-14 year olds presenting between 10 am and 10 pm. RESULTS: Of 10 175 assessed children, 204 aroused suspicion of child maltreatment (95% CI 163 to 214 per 10,000). In a logistic regression model of suspected maltreatment statistically significant associations were found with socioeconomic disadvantage, children living in single parent families, and developmental delay. There was no correlation with pre-school age, male gender, foreign origin, or living in urban areas. CONCLUSIONS: Child maltreatment based on immediate scoring of suspicion, focused on observable harm, occurred in 2% of a representative sample of paediatric emergency consultations in Italy. This was more common if there were associated social and developmental vulnerabilities. True prevalence of child maltreatment in emergency departments remains elusive because of changing definitions and forensic validation problems. PMID- 15781936 TI - Natural course of autoimmune thyroiditis in type 1 diabetes: association with gender, age, diabetes duration, and puberty. AB - AIMS: To investigate the natural history and incidence of autoimmune thyroiditis (AIT) in paediatric patients with type 1 diabetes (T1D). METHODS: Since 1990, annual screening for thyroid disease has been performed in children and adolescents with T1D. Antibodies against thyroperoxidase (anti-TPO) and thyroglobulin (anti-TG) as well as TSH were measured in 659 patients (54.3% boys). In 126 patients, anti-TPO and anti-TG levels were followed at yearly intervals from onset up to five years of T1D. Anti-TPO above 30 U/ml and anti-TG above 20 U/ml were considered positive, values above 100 U/ml as significantly raised and indicative of AIT. L-thyroxine treatment was started if TSH was higher than 4.5 microU/ml and/or thyroid gland enlargement on thyroid ultrasound was present. RESULTS: At initial screening, 15.4% of patients had raised anti-TPO and 14.4% anti-TG. Girls had more frequently raised antibodies than boys. Sixty two patients (9.4%, 61% girls) required treatment with L-thyroxine. The cumulative incidence (SE) of AIT after 10 years of diabetes was 0.14 (0.02), being significantly higher in females (0.18 (0.03)), particularly after the age of 12 years. At T1D onset, positive anti-TPO and anti-TG were present in 21 of 126 patients (16.7%), each. All patients with significantly increased values of anti TPO (n = 17, 148-5340 U/ml) and anti-TG (n = 11, 140-2000 U/ml) at T1D onset remained positive during the following five years. CONCLUSIONS: For early detection of autoimmune thyroiditis in children with T1D, measurement of anti-TPO and TSH at T1D onset and in yearly intervals after the age of 12 years is recommended. PMID- 15781939 TI - Large outbreak of measles in London: reversal of health inequalities. PMID- 15781937 TI - Identifying newborns at risk of significant hyperbilirubinaemia: a comparison of two recommended approaches. AB - AIMS: To compare the predictive performance of clinical risk factor assessment and pre-discharge bilirubin measurement as screening tools for identifying infants at risk of developing significant neonatal hyperbilirubinaemia (post discharge total serum bilirubin (TSB) >95th centile). METHODS: Retrospective cohort study of term and near term infants born in an urban community teaching hospital in Pennsylvania (1993-97). A clinical risk factor scoring system was developed and its predictive performance compared to a pre-discharge TSB expressed as a risk zone on a bilirubin nomogram. Main outcome measures were prediction model discrimination, range of predicted probabilities, and sensitivity, specificity, positive and negative predictive values, and likelihood ratios for various positivity criteria. RESULTS: The clinical risk factor scoring system developed included birth weight, gestational age <38 weeks, oxytocin use during delivery, vacuum extraction, breast feeding, and combination breast and bottle feeding. The pre-discharge bilirubin risk zone had better discrimination (c = 0.83; 95% CI 0.80 to 0.86) than the clinical risk factor score (c = 0.71; 95% CI 0.66 to 0.76) and predicted risk of significant hyperbilirubinaemia as high as 59% compared with a maximum of 44% for the clinical risk factor score. Neither the risk score nor the pre-discharge TSB risk zone predicted the outcome with > or =0.98 sensitivity without significantly compromising specificity (0.13 and 0.21, respectively). Multi-level clinical risk factor scores and TSB risk zones produced likelihood ratios of 0.15-3.25 and 0.05-9.43, respectively. CONCLUSIONS: The pre-discharge bilirubin expressed as a risk zone on an hour specific bilirubin nomogram is more accurate and generates wider risk stratification than a clinical risk factor score. PMID- 15781938 TI - Chemokine response in children with SARS. AB - The chemokine response of eight children with serologically confirmed severe acute respiratory syndrome (SARS) was longitudinally monitored. All had raised plasma interferon gamma inducible protein (IP-10) concentrations, which suggested an active type 1 T-helper lymphocyte mediated immune response. High circulating IP-10 levels could facilitate viral clearance and might play a role in assisting the recovery of the patients. PMID- 15781940 TI - Congenital cutaneous candidiasis with funisitis. PMID- 15781941 TI - Recent advances in congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is a common birth defect which continues to challenge paediatric surgeons and intensivists. Affecting approximately 1:2500 births, a baby with CDH is born every 24-36 hours in the UK. PMID- 15781943 TI - Our life with Jonathan. PMID- 15781942 TI - Early programming of adult diseases in resource poor countries. AB - Considerable evidence now exists to suggest that early exposure to nutritional deprivation can have long term consequences to health, with low birth weight now considered a risk factor for later health outcomes such as coronary heart disease, stroke, type 2 diabetes, and the metabolic syndrome. Of importance, such effects are most exaggerated when faced with over-nutrition in later life, forming the basis for the "thrifty phenotype" hypothesis. The evidence in support of these associations comes largely from retrospective cohort studies in which adult outcomes were correlated with birth weight records. Relatively little data is available from developing countries, where long term record keeping of birth weight data has not been a high priority. Arguably however, such countries are at the greatest risk from the mismatch of early nutritional deprivation and later nutritional affluence. This paper explores the importance of the "developmental origins of health and disease" hypothesis in resource poor countries. PMID- 15781944 TI - Risk of life threatening apnoea after immunisation. PMID- 15781945 TI - NSF for children and young people in Wales. PMID- 15781946 TI - Maternal vitamin D deficiency, refractory neonatal hypocalcaemia, and nutritional rickets. PMID- 15781947 TI - Addressing child welfare concerns: a new approach. PMID- 15781948 TI - Apnoeas in bronchiolitis: is there a role for caffeine? PMID- 15781949 TI - Uterine cancer after use of clomiphene citrate to induce ovulation. AB - Clomiphene citrate, a selective estrogen receptor modulator, increases estradiol levels and consequently may increase risk of cancer of the uterine corpus. The authors conducted a retrospective cohort study of 8,431 US women (145,876 woman years) evaluated for infertility during 1965-1988. Through 1999, 39 uterine cancers were ascertained by questionnaire or cancer and death registries. Poisson regression estimated adjusted rate ratios. Study results suggest that clomiphene may increase uterine cancer risk (rate ratio (RR) = 1.79, 95% confidence interval (CI): 0.9, 3.4) and present evidence of a dose response (p(trend) = 0.07) and latency effect (p(trend) = 0.04). Uterine cancer risk increased with clomiphene dose (RR = 1.93, 95% CI: 0.9, 4.0 for >900 mg), menstrual cycles of use (RR = 2.16, 95% CI: 0.9, 5.2 for >or=6 cycles), and time elapsed since initial use (RR = 2.50, 95% CI: 0.9, 7.2 for women followed for >or=20 years). Risk was more strongly associated with clomiphene among nulligravid (RR = 3.49, 95% CI: 1.3, 9.3) and obese (RR = 6.02, 95% CI: 1.2, 30.0) women, with risk substantially elevated among women who were both obese and nulligravid (RR = 12.52, 95% CI: 1.5, 108.0). Clomiphene may increase uterine cancer risk, with higher doses leading to higher risk. Long-term follow-up of infertility cohorts is necessary to clarify the association between clomiphene use and uterine cancer. PMID- 15781950 TI - Use of antibiotics and risk of breast cancer. AB - A recent nested case-control study found that increasing use of antibiotics was associated with a significantly elevated risk of breast cancer. The authors attempted to replicate this finding with a similar study design using the General Practice Research Database in the United Kingdom. Women aged 30-79 years who were registered in the database between January 1995 and December 2001 comprised the study cohort. A total of 3,708 women with incident cases of breast cancer and 20,000 frequency-matched controls were entered into a nested case-control analysis. Use of antibiotics was not associated with an increased risk of breast cancer. For categories of increasing cumulative days of use (1-50, 51-100, 101 500, and >or=501 days), the corresponding odds ratios were 1.0 (95% confidence interval (CI): 0.9, 1.1), 1.0 (95% CI: 0.8, 1.1), 0.9 (95% CI: 0.7, 1.0), and 1.2 (95% CI: 0.9, 1.6) (p = 0.31 for trend). On the basis of these results, antibiotic use does not appear to be a major determinant of breast cancer risk. PMID- 15781951 TI - Effect of sunscreen and clothing on the number of melanocytic nevi in 1,812 German children attending day care. AB - The number of melanocytic nevi is the most important risk factor for cutaneous melanoma. This 1998 cross-sectional study of 1,812 children aged 2-7 years from 78 day-care centers in Germany analyzed the protective effect of sunscreen and clothing on the number of melanocytic nevi. Total body nevi were counted, and pigmentary features were assessed. Parents underwent a standardized interview concerning their children's sun exposure and sun-protective behavior. Almost 95% of the children had used sunscreen previously. Children who used sunscreen and wore more clothing spent significantly longer periods on holidays in sunny climates (p < 0.001 and p = 0.006, respectively) and had a higher outdoor activity score at home (p < 0.001 and p = 0.012, respectively). Multivariate analysis adjusted for confounders showed no significant protective effects of applying sunscreen. Examination of the potential protective effect of clothing revealed an inverse dose-response correlation between the number of clothes worn at the beach or outdoor swimming pool and the number of melanocytic nevi (p < 0.001, adjusted for confounding). Randomized controlled trials are needed to confirm these results. Meanwhile, public education should aim to protect children primarily by sun avoidance and protective clothing, while sunscreen should also be used. PMID- 15781952 TI - A prospective study of hypertension and risk of uterine leiomyomata. AB - Although uterine leiomyomata (also known as fibroids or myomas) affect the reproductive health and well-being of approximately 25% of premenopausal women, risk factors are poorly understood. Elevated diastolic blood pressure may increase fibroid risk through uterine smooth muscle injury, not unlike atherosclerosis. The authors prospectively examined the relation between diastolic blood pressure and incidence of clinically detected leiomyomata. The sample included 104,233 premenopausal nurses from 14 US states enrolled in the Nurses' Health Study II. Participants, aged 25-42 years, had intact uteri and no history of cancer or fibroids at enrollment in 1989. During the 827,348 woman years of follow-up (1989-1999), 7,466 incident diagnoses of uterine leiomyomata, confirmed by ultrasound or hysterectomy, were reported. With a multivariable Cox proportional hazards model, the relative risk of self-reported ultrasound- or hysterectomy-confirmed uterine leiomyomata according to diastolic blood pressure in 1989 and time-varying antihypertensive use was estimated. With adjustment for age, race/ethnicity, body mass index, and reproductive history covariates, for every 10-mmHg increase in diastolic blood pressure, the risk of fibroids rose 8% (5-11%) and 10% (7-13%) among nonusers and users of antihypertensive medications, respectively. Elevated blood pressure has an independent, positive association with risk for clinically detected uterine leiomyomata among premenopausal women. Investigating this association may suggest possible pathways to prevent fibroids. PMID- 15781954 TI - Depressive symptoms and mortality among persons with and without diabetes. AB - Although people with diabetes mellitus have a high risk of depression and depression may increase mortality among people with other conditions, the impact of depression on mortality risk among people with diabetes needs further examination. Using survival analysis, the authors analyzed longitudinal data from the NHANES I Epidemiologic Follow-up Study (1982-1992). The findings showed that the presence of severe depressive symptoms significantly elevated mortality risk among US adults with diabetes; the same pattern was not observed among people without diabetes. After results were controlled for sociodemographic, lifestyle, and health-status variables, diabetic persons with Centers for Epidemiologic Studies Depression (CES-D) Scale scores of 16 or more had 54% greater mortality than those with scores under 16 (p = 0.004). After exclusion of participants who died during the first year of follow-up, mortality remained higher among those with CES-D scores greater than or equal to 22 as compared with those with CES-D scores less than 16, but not among those with CES-D scores between 16 and 21. No significant relation between depression and mortality was found in the nondiabetic population. This analysis indicates that diabetes modifies the effect of depression on mortality. It also demonstrates the importance of observing subgroups, rather than aggregated populations, when examining the effect of depression on mortality. PMID- 15781953 TI - Physical activity, APOE genotype, and dementia risk: findings from the Cardiovascular Health Cognition Study. AB - Physical activity may help preserve cognitive function and decrease dementia risk, but epidemiologic findings are inconsistent. The authors conducted a prospective study to determine the association between physical activity and risk of dementia, Alzheimer's disease, and vascular dementia. The US study population comprised 3,375 men and women aged 65 years or older, free of dementia at baseline, who participated in the Cardiovascular Health Cognition Study in 1992 2000. Leisure-time energy expenditure and an activity index reflecting number of different physical activities were calculated. Analyses were based on Cox proportional hazards models. There were 480 incident cases of dementia over an average of 5.4 years of follow-up. After multivariate adjustment, participants in the highest quartile of physical energy expenditure had a relative risk of dementia of 0.85 (95% confidence interval: 0.61, 1.19) compared with those in the lowest quartile, and participants engaging in >or=4 activities had a relative risk of dementia of 0.51 (95% confidence interval: 0.33, 0.79) compared with those engaging in 0-1 activity. These associations were more marked in apolipoprotein E genotype (APOE) epsilon4 allele noncarriers but were absent in carriers. A similar pattern was observed for Alzheimer's disease and vascular dementia. Mechanisms to explain the observed relations deserve further study. PMID- 15781955 TI - Risk factors for sternal and leg surgical site infections after cardiac surgery in Taiwan. AB - To account for time factors related to hospitalization, the authors calculated incidence rates of surgical site infection (SSI) instead of cumulative incidence and assessed risk factors for SSI after cardiac surgery. From July 1999 to August 2000, all cardiac surgery patients in the Taipei Veterans General Hospital (Taipei, Taiwan) were invited to join the study. Data were collected by chart review, including information on patient characteristics and potential risk factors at the preoperative, intraoperative, and postoperative stages. The authors employed multiple logistic regression analyses using a generalized logit model to assess associations of interest. SSI incidence rates were 2.5 episodes per 1,000 person-days for the sternum and 3.6 episodes per 1,000 person-days for the leg. After adjustment for covariates, age (in years), gender (female vs. male), New York Heart Association classification (moderate/severe vs. normal/mild), creatinine concentration (mg/dl), and duration of surgery (in minutes) were significantly associated with sternal SSI, while age, peripheral arterial occlusive disease, and length of stay in the intensive care unit (in hours) were significantly associated with leg SSI. In addition to patients' characteristics and health situations, the significant findings for duration of surgery and length of intensive care unit stay indicate that the incidence rate is more appropriate than cumulative incidence for studying risk factors for SSI. PMID- 15781956 TI - Dietary fat intake and risk of coronary heart disease in women: 20 years of follow-up of the nurses' health study. AB - The authors examined the associations of dietary fat and specific types of fat with risk of coronary heart disease (CHD) among 78,778 US women initially free of cardiovascular disease and diabetes in 1980. They documented 1,766 incident CHD cases (including 1,241 nonfatal myocardial infarctions and 525 CHD deaths) during 20 years of follow-up. Polyunsaturated fat intake was inversely associated with CHD risk (multivariate relative risk (RR) for the highest vs. the lowest quintile = 0.75, 95% confidence interval (CI): 0.60, 0.92; p(trend) = 0.004), whereas trans-fat intake was associated with an elevated risk of CHD (RR = 1.33, 95% CI: 1.07, 1.66; p(trend) = 0.01). The associations between intakes of polyunsaturated fat and trans-fat with CHD risk were most evident among women younger than age 65 years (for polyunsaturated fat, RR = 0.66, 95% CI: 0.50, 0.85; p(trend) = 0.002 and for trans-fat, RR = 1.50, 95% CI: 1.13, 2.00; p(trend) = 0.01). The inverse association between polyunsaturated fat intake and CHD risk was strongest among women whose body mass index was >or=25 kg/m(2). Findings continue to support an inverse relation between polyunsaturated fat intake and CHD risk, particularly among younger or overweight women. In addition, trans-fat intake was associated with increased risk of CHD, particularly for younger women. PMID- 15781957 TI - Educational level and osteoporosis risk in postmenopausal Chinese women. AB - Many studies have shown that better education is favorable for lowering the risks for a number of chronic diseases, but little information is available on the relation with bone health. The authors examined the association of educational level, classified as levels I-IV, with bone mineral density (BMD) and with the prevalence of osteoporosis among 685 population-based, postmenopausal, Chinese women aged 48-63 years during 1999-2001. They observed a significant dose response positive relation between educational level and BMDs at the total-body (p = 0.011), lumbar spine, and hip sites (p < 0.001) after adjusting for age, years since menopause, and body weight. Mean BMDs of educational level IV women were higher by 4.2-11.9% at the various sites compared with level I women (analysis of covariance, p < 0.05). Similarly, the authors also observed a significant inverse dose-response relation between educational level and prevalence of osteoporosis. Women of educational level I were 3.5-8.6 times more likely to be osteoporotic compared with those of level IV at the various sites. The proportion of BMD or osteoporosis variations accounted for by educational level was attenuated by about 40% after further controlling for potentially explanatory covariates. In conclusion, a higher level of education is independently associated with better BMDs and lower prevalence of osteoporosis among postmenopausal Chinese women. PMID- 15781958 TI - Association of serum cholesterol and history of school suspension among school age children and adolescents in the United States. AB - The dietary guidelines developed for adults have been extended to children, but the role of serum cholesterol in the neurodevelopment of children is poorly understood. In the Third National Health and Nutrition Examination Survey (1988 1994), serum total cholesterol was measured in 4,852 children aged 6-16 years. Psychosocial development was evaluated by interviewing the mother regarding the child's history of school suspension or expulsion and difficulty in getting along with others. After adjustment for family socioeconomic status, maternal marital status and education, children's nutrition, and academic performance, the odds ratios of children with various concentrations of total cholesterol showed the children to be equally comfortable in their own peer subculture and not to be different in the proportion that had seen a mental health professional. However, non-African-American children with a serum total cholesterol concentration below the 25th percentile (<145 mg/dl) were almost threefold more likely to have been suspended or expelled from schools than their peers with total cholesterol at or above the 25th percentile (odds ratio = 2.96, 95% confidence interval: 1.55, 5.64). The authors concluded that, among non-African-American children, low total cholesterol is associated with school suspension or expulsion and that low total cholesterol may be a risk factor for aggression or a risk marker for other biologic variables that predispose to aggression. PMID- 15781959 TI - A chain multinomial model for estimating the real-time fatality rate of a disease, with an application to severe acute respiratory syndrome. AB - It is well known that statistics using cumulative data are insensitive to changes. World Health Organization (WHO) estimates of fatality rates are of the above type, which may not be able to reflect the latest changes in fatality due to treatment or government policy in a timely fashion. Here, the authors propose an estimate of a real-time fatality rate based on a chain multinomial model with a kernel function. It is more accurate than the WHO estimate in describing fatality, especially earlier in the course of an epidemic. The estimator provides useful information for public health policy makers for understanding the severity of the disease or evaluating the effects of treatments or policies within a shorter time period, which is critical in disease control during an outbreak. Simulation results showed that the performance of the proposed estimator is superior to that of the WHO estimator in terms of its sensitivity to changes and its timeliness in reflecting the severity of the disease. PMID- 15781961 TI - No go for brain tumors? AB - The Nogo gene and its products are well known as adult central nervous system (CNS) myelin inhibitors of neuronal regeneration. We review here experimental findings that might link Nogo to CNS malignancy. These links are founded on two very different modes of cellular action by Nogo isoforms. Acting intracellularly and in conjunction with other molecules, cytoplasmic domains of Nogo might predispose cancer cells to apoptotic susceptibility. On the other hand, extracellular domains of Nogo might inhibit the migration and invasion of CNS tumors. Depending on the physiological context, Nogo isoforms might therefore be antitumorigenic or have tumor-suppressing activities. PMID- 15781963 TI - Orphanin FQ antagonizes the inhibition of Ca(2+) currents induced by mu-opioid receptors. AB - Orphanin FQ (OFQ), an endogenous peptide ligand of opioid receptor-like receptors (ORLs), has properties similar to traditional opioids. This peptide inhibits adenylyl cyclase and voltage-gated calcium channels but stimulates inwardly rectifying potassium channels. Among other actions, however, OFQ also has pharmacological functions that are different from, or even opposite to, those of opioids. For example, OFQ antagonizes the behavioral analgesic effects mediated by kappa- and mu-opioid receptors. In a previous paper, we reported that OFQ antagonizes inhibition of calcium channels mediated by kappa-opioid receptors. We report here that OFQ also antagonizes the inhibition of calcium channels mediated by mu-opioid receptor. Further, single-cell RT-PCR reveals that the antagonistic effect of OFQ is correlated with the presence of ORL1 mRNA in individual cells. PMID- 15781964 TI - Bromocriptine and clozapine regulate dopamine 2 receptor gene expression in the mouse striatum. AB - In a previous study, we showed that the psychoactive drug caffeine alters the expression of the dopamine 2 receptor (D2R) gene in vitro and in vivo. Here, we report that acute administration of antipsychotic and anti-parkinsonian drugs also regulate D2R gene expression in PC12 cells and in the mouse striatum. Treatment of PC12 cells with the atypical antipsychotic and specific 5-HT antagonist clozapine (60 microM) reduced D2R/luciferase reporter expression by 46% after 24 h. However, male and female mice treated with a clinical dose of clozapine (10 mg/kg) showed no changes in striatal D2R mRNA expression when assayed by quantitative RT-PCR. Treatment of PC12 cells with the specific D2R agonist anti-parkinsonian drug, bromocriptine mesylate (BCM; 5 microM) also resulted in decreased D2R/luciferase reporter activity (27%). In contrast to clozapine, a clinical dose of BCM (16 mg/kg) led to a 21% decrease and a 45% increase in striatal D2R mRNA expression in male and female mice, respectively, after 24 h. Coadministration of clozapine and BCM in PC12 cells resulted in a synergistic decrease in D2R/luciferase reporter expression (68%), and coadministration of these drugs in vivo led to decreases in striatal D2R mRNA expression in both male and female mice (45% and 22%, respectively). Collectively, these results indicate that clozapine, BCM, or a combination of these drugs have differential effects on dopamine receptor gene expression and might also affect striatal physiology in a sexually dimorphic manner. PMID- 15781962 TI - Kinetic modeling of Na(+)-induced, Gbetagamma-dependent activation of G protein gated K(+) channels. AB - G protein-activated K(+)(GIRK) channels are activated by numerous neurotransmitters that act on Gi/o proteins, via a direct interaction with the Gbetagamma subunit of G proteins. In addition, GIRK channels are positively regulated by intracellular Na(+) via a direct interaction (fast pathway) and via a GGbetagamma-dependent mechanism (slow pathway). The slow modulation has been proposed to arise from the recently described phenomenon of Na(+)-induced reduction of affinity of interaction between GalphaGDP and Gbetagamma subunits of G proteins. In this scenario, elevated Na(+) enhances basal dissociation of G protein heterotrimers, elevating free cellular Gbetagamma and activating GIRK. However, it is not clear whether this hypothesis can account for the quantitative and kinetic aspects of the observed regulation. Here, we report the development of a quantitative model of slow, Na(+)-dependent, G protein-mediated activation of GIRK. Activity of GIRK1F137S channels, which are devoid of direct interaction with Na(+), was measured in excised membrane patches and used as an indicator of free GGbetagamma levels. The change in channel activity was used to calculate the Na(+)-dependent change in the affinity of G protein subunit interaction. Under a wide range of initial conditions, the model predicted that a relatively small decrease in the affinity of interaction of GalphaGDP and GGbetagamma (about twofold under most conditions) accounts for the twofold activation of GIRK induced by Na(+), in agreement with biochemical data published previously. The model also correctly described the slow time course of Na(+) effect and explained the previously observed enhancement of Na(+)-induced activation of GIRK by coexpressed Galphai3. This is the first quantitative model that describes the basal equilibrium between free and bound G protein subunits and its consequences on regulation of a GGbetagamma effector. PMID- 15781967 TI - Young investigator award recipient. AB - Dr. Zhongcong Xie started his research career in 1989 with graduate studies in behavioral pharmacology at Wayne State University, Detroit, Michigan.He moved to Harvard Medical School, Boston, Massachusetts in 1994 as a postdoctoral research fellow in both human and animal behavioral research. After years of research training, he returned to the clinical arena in 1997 as an Anesthesiology resident at Massachusetts General Hospital and Harvard Medical School. After completing anesthesia residency training, he decided to become a physician-scientist and continue his research in neuroscience. He joined the Genetics and Aging Research Unit at Massachusetts General Hospital and Harvard Medical School supervised by Dr. Rudolph Tanzi in the year 2000 to study Alzheimer's disease. His research on the neuropathogenesis of Alzheimer's disease has focused primarily on the molecular mechanisms underlying beta-amyloid precursor protein processing and beta-amyloid protein production, especially regarding regulation of the gamma secretase complex.Additionally, his work has addressed the role of presenilin related apoptosis in Alzheimer's disease. PMID- 15781965 TI - A TASK3 channel (KCNK9) mutation in a genetic model of absence epilepsy. AB - Childhood absence epilepsy is an idiopathic, generalized, nonconvulsive epilepsy with a multifactorial genetic etiology. The KCNK9 gene coding for the TASK3 (Twik like acid-sensitive K or =800 nM). Spiking experiments of PCR reactions or CHO-K1 cells with the chimeraplast confirmed that the repair was not artifactual. However, when treated recombinant CHO cells were passaged for 10 d and then subcloned, no conversion could be detected when >90 clones were analyzed by locus-specific PCR-RFLP. We conclude that the apparent efficient repair of the APOE4 gene in CHO cells or lymphocytes 48 h post-treatment is unstable, possibly because the high levels of chimeraplast and polyethyleneimine that were needed to induce nucleotide substitution are cytotoxic. PMID- 15781971 TI - Dystonia-associated forms of torsinA are deficient in ATPase activity. AB - Early-onset dystonia is caused by mutations in the torsinA protein, a putative member of the AAA+ class of ATPases. In this study we have evaluated the ATPase activity of bacterially expressed wild-type torsinA and its disease-associated mutant forms. Upon overexpression in Escherichia coli, recombinant torsinA proteins were accumulated as insoluble inclusion bodies and required refolding to become soluble and catalytically active. The refolded wild-type and mutant torsinA proteins were capable of hydrolyzing ATP, but their specific ATPase activities differed significantly. Deletions of the amino acid residues E302/303 and F323-Y328 resulted in a decrease of ATPase activity to approximately 35% and approximately 75% of the wild-type level, respectively. ATPase activity of wild type and mutant torsinA proteins was influenced by factors that varied with cell stress, such as temperature, pH, and ionic strength, and was inhibited by sodium vanadate. Our results provide the first direct evidence for a role of torsinA as an active ATPase and suggest that the mutations in torsinA might affect normal functions of the protein by reducing its enzymatic activity. PMID- 15781972 TI - A distinctive molecular signature of multiple sclerosis derived from MALDI-TOF/MS and serum proteomic pattern analysis: detection of three biomarkers. AB - Although multiple sclerosis (MS) is an immune-mediated disorder, serological testing to aid in its diagnosis has not been developed. To test the hypothesis that the pathological changes in MS can be detected by analyzing a molecular signature of serum proteomic patterns, we tested sera from 25 newly diagnosed relapsing-remitting MS patients and 25 healthy controls with matrix-assisted laser desorption/ ionization time of flight (MALDI-TOF) mass spectrometry, following albumin depletion and desalting. Analysis of MALDI-TOF/mass spectrometry data, using proteomic spectral pattern recognition software, revealed a distinct proteomic pattern in the MS group determined by 3 biomarkers at 8687, 8773, and 8818 mass-to-charge ratios. Although our data are representative of analysis on a small number of samples and are preliminary, we conclude that MALDI-TOF/mass spectrometry, in combination with serum proteomic pattern analysis, could be useful in the diagnosis of MS, and a larger, masked trial to identify proteomic spectral patterns characteristic of relapsing remitting, primary progressive and secondary progressive variants of MS is justified. PMID- 15781973 TI - RNA interference technologies for understanding and treating neurodegenerative diseases. AB - RNA interference (RNAi) is an evolutionarily conserved process that silences gene expression through double-stranded RNA species in a sequence-specific manner. With the completion of genome sequencing in multiple organisms, RNAi provides an efficient reverse genetics tool to reveal gene functions on a genome-wide scale. Conditional/inducible RNAi offers a new way to analyze gene function at different developmental stages and to create a new generation of animal models of human diseases. The sequence-specificity of RNAi and the fact that it is a naturally occurring process in human make it an excellent therapeutic tool for a wide range of diseases. This article provides a brief review of the current understandings of the mechanism of RNAi and its application to the nervous system, with particular focus on its application to understand mechanisms of neurodegenerative diseases. The prospects of the application of RNAi in clinical setting to treat these devastating diseases will also be presented. PMID- 15781974 TI - Gene-targeting technologies for the study of neurological disorders. AB - Studies using genetic manipulations have proven invaluable in the research of neurological disorders. In the forefront of these approaches is the knockout technology that engineers a targeted gene mutation in mice resulting in inactivation of gene expression. In many cases, important roles of a particular gene in embryonic development have precluded the in vivo study of its function in the adult brain, which is usually the most relevant experimental context for the study of neurological disorders. The conditional knockout technology has provided a tool to overcome this restriction and has been used successfully to generate viable mouse models with gene inactivation patterns in certain regions or cell types of the postnatal brain. This review first describes the methodology of gene targeting in mice, detailing the aspects of designing a targeting vector, introducing it into embryonic stem cells in culture and screening for correct recombination events, and generating chimeric and null mutant mice from the positive clones. It then discusses the special issues and considerations for the generation of conditional knockout mice, including a section about approaches for inducible gene inactivation in the brain and some of their applications. An overview of gene-targeted mouse models that have been used in the study of several neurological disorders, including Alzheimer's disease, Parkinson's disease, Huntington's disease, seizure disorders, and schizophrenia, is also presented. The importance of the results obtained by these models for the understanding of the pathogenic mechanism underlying the disorders is discussed. PMID- 15781975 TI - Gene expression profiling in neurological disorders: toward a systems-level understanding of the brain. AB - Neurological disease arises when a sufficient number of neural cells stop performing their normal functions, lose their ability to respond to local environment, and die. In the last decade, a major technological leap led to the development of efficient and cost-effective high-throughput methods for determining gene expression. This, in turn, resulted in the rapid accumulation of data describing gene expression patterns in the brain of humans and experimental animals. This article reviews several of the most commonly used technologies to retrieve transcriptional information, such as large-scale expressed sequence tag (EST) sequencing, serial analyses of gene experiment, and different DNA microarray platforms. In addition, efforts focusing on data collection, sharing, and standardization are described. A section is devoted to the latest advances in large-scale gene expression in several neurological disorders, including Alzheimer's disease, Parkinson's disease, multiple sclerosis, and epilepsy. Finally, the need for integration of different sources of data is discussed in the context of systems biology and how such integration could result in improved diagnostics, therapies, and disease prevention. PMID- 15781980 TI - Steroidogenic acute regulatory protein: its presence and function in brain neurosteroidogenesis. AB - Neurosteroids are synthesized de novo and involved in a variety of physiological functions in the central and peripheral nervous systems. Although the steroidogenic acute regulatory protein (StAR) plays an essential role in the steroidogenesis of peripheral endocrine glands, its presence and role in the brain had been previously questioned because of difficulties in detecting it. However, a number of recent studies have confirmed the presence of StAR in rodent and human brains. Moreover, there is evidence suggesting that StAR plays a role in steroidogenesis in the brain, as it does in peripheral endocrine organs. The present review presents data regarding the presence and role of StAR in brain steroidogenesis, demonstrating the essential characteristics of the protein. PMID- 15781976 TI - Calcium-regulated signaling pathways: role in amyloid beta-induced synaptic dysfunction. AB - Amyloid beta (Abeta) peptides have been shown to impair synaptic function, especially long-term synaptic plasticity, in transgenic mouse models of Alzheimer's disease (AD) and in acute hippocampal preparations. In the transgenic mice overexpressing mutant forms of human amyloid precursor protein (APP), the deficits in hippocampal long-term potentiation (LTP) occur prior to synaptic loss and cell death, suggesting early functional changes at these synapses. Recent studies demonstrate that Abeta-induced synaptic dysfunction is linked with altered Ca2+ signaling in hippocampal neurons. While reducing Ca2+ influx through NMDA receptors, Abeta peptides elevate intracellular Ca2+ concentration by enhancing Ca2+ influx from voltage-gated Ca2+ channels or nonselective cation channels, or by stimulating Ca2+ release from intracellular stores. Interestingly, acute application of Abeta or APP overexpression inhibits activity dependent regulation of several protein kinase pathways that require Ca2+ influx via NMDA receptors for activation, including Ca2+/calmodulin-dependent protein kinase II, protein kinase A, and extracellular regulated kinases (Erk). On the other hand, activation of Ca2+-dependent protein phosphatase 2B (calcineurin) is implicated in Abeta inhibition of LTP. Thus, multiple Ca2+-regulated signaling pathways are involved in the synaptic action of Abeta, and malfunction of these pathways may underlie the synaptic dysfunction in early AD. PMID- 15781977 TI - Techniques for brain imaging in vivo. AB - Brain imaging technologies provide an unparalleled tool to assess the physiological and pathological changes in the central nervous system. The aim of this work is to review the technical characteristics of widely used imaging technologies, such as magnetic resonance imaging (MRI), positron emission tomography (PET), or single photon emission computed tomography (SPECT), as well as their utility in clinical diagnosis and follow-up of different neurological disorders. High-resolution developments of MRI and PET, as well as the more recent imaging technology, multiphoton microscopy, for small animal imaging are also analyzed, as rodent models offer an enormous opportunity to study not only the underlying mechanisms of Alzheimer's disease, Parkinson's disease, depression, and schizophrenia among others, but also potential new treatments for all these diseases. PMID- 15781981 TI - Hormonal regulation and localization of estrogen, progestin and androgen receptors in the endometrium of nonhuman primates: effects of progesterone receptor antagonists. AB - This article reviews the effects of estradiol (E(2)), progesterone (P) and P receptor antagonists (PA) on the rhesus macaque endometrium. Ovariectomized macaques can be treated with implants of estradiol (E(2)) and P to induce precisely controlled, artificial menstrual cycles. During these cycles, treatment with E(2) alone induces an artificial proliferative phase marked by extensive endometrial epithelial cell proliferation and increased expression of stromal and epithelial estrogen receptor (ER) and P receptor (PR). Androgen receptor (AR) is also upregulated by E(2) but is expressed only by the endometrial stroma. Progesterone acts on the E(2) primed endometrium to induce secretory differentiation and causes suppression of epithelial and stromal ER, epithelial PR, and stromal AR in the functionalis zone. However, epithelial ER and PR are retained in the basalis zone during the secretory phase. When potent P antagonists (PA) are administered acutely at the end of an E(2) + P induced cycle, menses typically ensues similar to P withdrawal at the end of the menstrual cycle. When PAs are administered chronically there is significant blockage of all P- dependent effects including upregulation of ER, PR and AR and suppression of glandular secretory function. However, chronic PA administration also inhibits estrogen-dependent endometrial cell proliferation and growth. This endometrial antiproliferative effect is the basis of the clinical use of PA to control various diseases such as endometriosis. PMID- 15781982 TI - Steroid receptors in blood vessels of the rhesus macaque endometrium: a review. AB - Estradiol (E) and progesterone (P) act on the primate endometrium to induce dramatic changes in the vascular system during the menstrual cycle. These changes include vessel breakdown and bleeding during menses, heightened angiogenesis during the early proliferative phase, and extensive growth of the spiral arteries in the luteal phase of the cycle. Because steroid hormone action is dependent upon the presence of specific nuclear receptors in target tissues, we used immunocytochemistry with receptor-specific monoclonal antibodies to characterize the spatial and temporal expression of estrogen receptor alpha (ERalpha), estrogen receptor beta (ERbeta), progesterone receptor PR and androgen (A) receptor (AR) in the endometrial vessels of rhesus macaques (Macaca mulatta). The only sex steroid receptor that was present in the endothelium and smooth muscle walls of endometrial vessels was ERbeta. ERalpha, PR, and AR were not detectable in either the endothelium or vascular smooth muscle cells of primate endometrial vessels. However, all of these receptors were strongly expressed by the perivascular stroma, and in these cells, all were modulated by the changes in levels of E and P during the cycle. We concluded that any direct effects of E on endometrial vessels would be mediated by ERbeta, and that the actions of P and A, and possibly some of E, were indirectly mediated through perivascular stromal cells. PMID- 15781983 TI - Role of stromal-epithelial interactions in hormonal responses. AB - Steroid sex hormones (17beta-estradiol, testosterone, dihydrotestosterone, and progesterone) and aryl hydrocarbons such as the dioxins regulate epithelial proliferation and secretory protein production and differentiation in their respective target organs in male and female urogenital tracts and mammary glands. Recent evidence has demonstrated that stromal-epithelial interactions are critical for mediating the effects of these molecules on epithelial cells. Our results have indicated that estradiol, testosterone, progesterone, and dioxin regulate epithelial proliferation (stimulation or inhibition) via paracrine mechanisms requiring the appropriate receptor in the stroma. The androgen receptor (AR), estrogen receptor alpha (ERalpha), progesterone receptor (PR), or aryl hydrocarbon receptor (AhR) in the epithelium are neither necessary nor sufficient for the regulation of epithelial proliferation. Moreover, during prostatic development, signaling through the stromal AR is required to induce prostatic epithelial identity, ductal morphogenesis and glandular differentiation. Epithelial functional differentiation is regulated in the prostate, uterus, and vagina via AR (prostate) and ERalpha(uterus and vagina). In these organs both epithelial and stromal steroid receptors are required for steroidal regulation of certain aspects of epithelial differentiation such as epithelial secretory protein production in the uterus and epithelial cornification in the vagina and prostate (squamous metaplasia). The mechanistic basis of these stromal-epithelial interactions is poorly understood, but growth factors appear to be mediators of these cell-cell interactions. PMID- 15781984 TI - Estrogen receptors and their downstream targets in cancer. AB - Estrogen has crucial roles in the proliferation of cancer cells in reproductive organs such as the breast and uterus. Estrogen-stimulated growth requires two estrogen receptors (ERalpha and ERbeta) which are ligand-dependent transcription factors. High expression of ERs is observed in a large population of breast tumors. In addition, the positive expression of ERs correlates with well differentiated tumors, a favorable prognosis, and responsiveness to an endocrine therapy with anti-estrogen drugs in patients with breast cancer. Transcription activities of ERs can be regulated by interacting proteins such as coactivators and kinases as well as ligand-binding. Moreover, ER isoforms lacking an ability to transactivate are involved in breast cancer. Downstream target genes of ERs have important roles in mediating the estrogen action in breast cancer. We have isolated and characterized several novel estrogen-responsive genes to clarify the molecular mechanism of the estrogen action in target cells. Among these genes, the estrogen-responsive finger protein (Efp) was found to be highly expressed in breast cancer. Efp as a ubiquitin ligase (E3) is involved in the proteasome dependent degradation of the 14-3-3sigma protein, one of cell cycle brakes, this degradation resulting in the promotion of breast cancer growth. A full understanding of the expression and function of ERs and their target genes could shed light on how estrogen stimulates the initiation and promotion of cancer, providing a new approach to diagnose and treat cancer. PMID- 15781985 TI - Immunohistochemical localization of the ACTH (MC-2) receptor in the rat placenta and adrenal gland. AB - The adrenocorticotropic hormone (ACTH) acts on adrenocortical cells and promotes steroidogenesis by specific binding to the ACTH (MC-2) receptor (ACTHR). To gain an insight into ACTH action on local steroidogenic organs, we examined the immunohistochemical expression of ACTHR in rat adrenal glands and placentas during the mid-late gestation period. Antibodies against synthetic ACTHR peptides were raised in rabbits, and Western blot analysis showed that the antibody reacted with specific proteins in the rat adrenal glands and placentas. The peroxidase-labeled antibody method revealed that ACTHR was distributed in the plasma membrane and cytoplasm of the parenchymal cells of the adrenocortical zona fasciculata. In the placenta, ACTHR was distributed in the junctional spongiotrophoblasts at day 13 of gestation--with a gradual decrease in the staining during the gestational period, whereas ACTHR appeared in the placental labyrinthine cells from days 15 to 19 of gestation. Immunoelectron microscopy revealed that ACTHR was also localized in the ribosomes of the fasciculata cells and the labyrinthine cells. Our findings suggest that ACTHR may play a physiological role in steroidogenesis in the adrenal cortical parenchymal cells as well as in the trophoblasts of rat placentas during mid-late gestation. PMID- 15781986 TI - Expression of keratinocyte growth factor and its receptor in human breast cancer: its inhibitory role in the induction of apoptosis possibly through the overexpression of Bcl-2. AB - Keratinocyte growth factor (KGF), a mesenchymal cell derived paracrine growth factor that regulates normal epithelial cell proliferation, appears to be an essential mediator of steroids in various reproductive organs. The present study was designed to determine the expression and role of KGF and its receptor (KGFR) in human breast carcinoma tissues by immunohistochemistry. We also compared the results with the expression of estrogen receptor alpha(ERalpha), ERbeta, the proliferative activity assessed by the labeling index (LI) for the Ki-67 antigen, apoptotic frequency assessed by terminal dUTP nick end-labeling (TUNEL) index, and the expression of Bcl-2. All of KGF-positive cases were ERalpha- positive (p<0.05), but not that of ERbeta, while all of KGFR-positive cases were ERbeta positive (p<0.05), but not that of ERalpha. The specimens with the coexpression of KGF and KGFR significantly correlated with a lower TUNEL index (p<0.05), but not with Ki-67 LI in breast cancer tissues. Further analysis at the cellular level revealed that Bcl-2 was colocalized in KGFR-positive cells, and these cells were almost negative for TUNEL staining. Bcl-2-positive cells were also associated with ERbeta, as expected. Therefore, the results indicate that ERalpha may be involved in KGF expression, and that the coexpression of KGF and KGFR may play an inhibitory role in the induction of apoptosis possibly through the up regulation of Bcl-2 expression in human breast cancer. PMID- 15781987 TI - Expression of the arylhydrocarbon receptor in the peri-implantation period of the mouse uterus and the impact of dioxin on mouse implantation. AB - The arylhydrocarbon receptor (AhR) is a nuclear transcription factor mediating toxic effects of chemicals such as dioxins. The 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD), a member of polyhalogenated aromatic hydrocarbons family, exerts a wide-variety of toxic effects in a tissue- and species-specific manner including the reproduction process. Recently, AhR-mediated direct effects of TCDD on a cell specific interaction with ovarian steroids have been shown. However, information regarding the effects of TCDD on the mouse implantation is limited. We therefore examined the expression and localization of AhR in the pregnant mouse uterus from 4 to 10 days of gestation (day 4 to day 10) using immunohistochemistry to investigate the effect of TCDD on uterine tissue during the peri-implantation period. Intense AhR expression was detected in the uterine vasculature throughout the periods examined. We also found that implanted blastocysts and their surrounding luminal epithelia and decidualized stroma expressed AhR on day 5. On days 6 and 7, persistent AhR expression was found in the transitional zone between the invading embryonic tissue and decidual tissue. On days 9 to 10, placental vasculature and spongiotrophoblasts displayed AhR immunoreactivity. The administration of TCDD on day 4 decreased the number of surviving implanted embryos on day 7 in a dose-dependent manner. This effect of TCDD was inhibited by the simultaneous administration of an AhR antagonist, alpha-naphthoflavone (alpha NF). The spatio-temporal expression of AhR during the peri-implantation phase of the mouse uterus may indicate functional roles of this orphan receptor in fetomaternal interactions as well as substantiate the risk of exposure to chemicals such as dioxins during the reproductive period. PMID- 15781988 TI - Proteasome function and pathological proteins in the pathogenesis of Parkinson's disease. PMID- 15781989 TI - Nitric oxide induces apoptosis in mouse C2C12 myoblast cells. AB - To investigate whether nitric oxide (NO) induces apoptosis in myoblast cells, the effect of the sodium nitroprusside (SNP), NO donor, on the apoptosis of mouse C2C12 myoblast cells was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay, flow cytometry, 4,6-diamidino-2 phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, and caspase-3 enzyme assay. Mouse C2C12 myoblast cells treated with SNP exhibited several apoptotic features. SNP increased p53 expression and bax expression. SNP also enhanced caspase-3 enzyme activity. The data show that NO may induce apoptotic cell death in myoblast cells through the activation of p53-, bax-, and caspase-dependent intracellular death-related pathways. PMID- 15781990 TI - Functional proteins involved in regulation of intracellular Ca(2+) for drug development: the extracellular calcium receptor and an innovative medical approach to control secondary hyperparathyroidism by calcimimetics. AB - Circulating levels of calcium ion (Ca(2+)) are maintained within a narrow physiological range mainly by the action of parathyroid hormone (PTH) secreted from parathyroid cells. Parathyroid cells can sense small fluctuations in plasma Ca(2+) levels by virtue of a cell surface Ca(2+) receptor (CaR) that belongs to the superfamily of G-protein-coupled receptors. Calcimimetics are positive allosteric modulators that activate the CaR on parathyroid cells and thereby immediately suppress PTH secretion. Pre-clinical studies with NPS R-568, a first generation calcimimetic compound, have demonstrated that daily oral administration inhibits the elevation of plasma PTH levels and parathyroid gland hyperplasia and ameliorates impaired bone qualities in rats with chronic renal insufficiency. The results of clinical trials with cinacalcet hydrochloride, a second generation calcimimetic compound, have shown that calcimimetics possess lowering effects not only on serum PTH levels but also on serum calcium x phosphorus product levels, a hallmark of an increased risk for cardiovascular death in dialysis patients with end-stage renal disease (ESRD). Thus, calcimimetics have considerable potential as an innovative medical approach to manage secondary hyperparathyroidism associated with ESRD. Indeed, cinacalcet hydrochloride has been approved in several countries and is the first positive allosteric modulator of any G protein-coupled receptor to reach the market. PMID- 15781991 TI - Involvement of the fas ligand and fas system in apoptosis induction of mouse uterine natural killer cells. AB - Uterine natural killer (uNK) cells in the pregnant uterus are known to be associated with the normal development of the placenta. In the mouse pregnant uterus, many uNK cells exist during mid pregnancy, although they show a sudden decrease during late pregnancy and almost disappear before delivery. Our previous study indicated that uNK cells showed clear apoptotic morphology during late pregnancy. Therefore, the present study was carried out to define the involvement of Fas ligand (FasL) and Fas in apoptosis induction of uNK cells. Immunohistochemical analyses revealed that uNK cells expressed FasL in the cytoplasmic granules and Fas on the cell membrane during late pregnancy. In lpr/lpr mice, which genetically lack Fas, many uNK cells were clearly observed during late pregnancy compared with wild-type mice, and moreover uNK cells still existed at day-18 of pregnancy, although there were few in wild-type mice during the same period. In the experiment of in vitro culture, uNK cells derived from wild-type placenta showed chromatin condensation and DNA fragmentation frequently following the anti-Fas antibody treatment, as compared with the control. From these results, it is suggested that FasL and Fas-dependent apoptosis regulates cell appearance of uNK cells in the mouse pregnant uterus. PMID- 15781992 TI - Diversity of Borrelia burgdorferi sensu lato in Russian Far East. AB - Thirty strains of Borrelia burgdorferi sensu lato have been isolated from Ixodes persulcatus ticks and from skin lesions of Lyme disease patients in the Russian Far East from 1997 to 2003. We amplified full-length outer surface protein A (ospA) gene of all strains. BLAST search and following phylogenetic analysis showed that strains form four well-defined groups. Four strains belong to Borrelia afzelii species. Other strains distributed into tree major groups, identified as Borrelia garinii. Indeed, based on the ospA gene comparison, phylogenetic relationship of these groups among each other does not differ from relationship among other previously defined groups inside B. burgdorferi sensu lato genogroup, such as B. afzelii or Borrelia bissettii. Further investigations of genetic and serologic properties of the strains belonging to those groups are required in order to clarify their taxonomic status. PMID- 15781993 TI - Experimental candidosis and recovery of Candida albicans from the oral cavity of ovariectomized rats. AB - The aim of this study was to analyze the development of candidosis and the recovery of C. albicans from the oral cavity of ovariectomized and sham ovariectomized rats. One hundred and twenty-four rats originally negative for Candida spp. in the oral cavity were divided into two groups: ovariectomized and sham-ovariectomized. Fifty-eight ovariectomized and the same quantity of sham ovariectomized rats were inoculated with C. albicans for the study of candidosis development and recovery of yeast. Four animals from each group were not inoculated with yeast suspension and were submitted to tongue dorsum morphologic analysis by optical and scanning electron microscopy. The development of candidosis in the tongue dorsum was observed by optical and scanning electron microscopy in the periods of 6 hr, 24 hr, 7 days and 15 days after the last inoculation. Recovery of C. albicans was performed by oral samples plating on Sabouraud agar after 1, 2, 5 and 7 days and progressively at each 15-day interval until negative cultures for yeasts were obtained. The results were analyzed by Mann-Whitney and Student's t tests. The tongue dorsum of sham-ovariectomized and ovariectomized rats, not infected by Candida, presented normal aspect. Among the infected rats, the ovariectomized group showed less occurrence of candidosis lesions and lower recovery of C. albicans from the oral cavity in relation to the sham-ovariectomized group. It could be concluded that candidosis was less frequent from the oral cavities of ovariectomized rats in relation to sham ovariectomized. PMID- 15781994 TI - Comparisons of the laccase gene among serotypes and melanin-deficient variants of Cryptococcus neoformans. AB - Cryptococcus neoformans is a fungus causing life-threatening infections in immunocompromised hosts. Melanin production is a major virulence factor of this fungus and the initial steps of dihydroxyphenylalanine (DOPA)-melanin biosynthesis pathways are catalyzed by laccase. To understand phylogenetic relationships among serotypes of three varieties, partial sequences (about 600 bases) of the laccase gene (CNLAC1) were determined in a total of 64 strains, including 10 melanin-deficient variants. The phylogenetic tree constructed from the nucleotide sequence grouped the 64 strains into the clusters corresponding to the three varieties. The diversity of the fragment sequences was very minor among strains of each of var. grubii and var. neoformans. Strains in var. gattii, however, were subdivided into two groups, although differences between serotypes B and C were not large. The sequences of the melanin-deficient variants were almost completely homologous to those of the melanin-producing strains in the same serotype. Results of laccase assay and northern blot analysis suggested that the lower melanin production in the variants was associated with lower transcription of the laccase gene. PMID- 15781995 TI - Preparation of monoclonal antibodies cross-reactive with orthopoxviruses and their application for direct immunofluorescence test. AB - Variola virus (smallpox virus), vaccinia virus (VV), cowpox virus (CPV) and ectromelia virus (EV) belong to the genus Orthopoxvirus of the family Poxviridae. To establish the possible diagnosis for smallpox infection, monoclonal antibodies (MAbs) against VV and CPV were produced. The cross-reactivity of seven MAbs with cells infected with various strains of the orthopoxviruses (CPV, VV and EV) was confirmed by an immunofluorescence (IF) test and other immunological analyses. Four and three MAbs reacted with the common antigen of all poxviruses (probably NP antigen) and the antigen involved in neutralization, respectively. We developed the IF test using these MAbs. The direct IF test required only 45 min to perform. Smallpox infection is now eradicated, but it is important to prepare for the diagnosis of smallpox in an emergency. The direct IF assay using MAbs cross-reactive with orthopoxviruses is rapid, simple, specific, applicable for multiple samples, and will make it possible to screen for and detect orthopoxviruses that include variola virus with tissue impression smears from skin lesions in most laboratories or institutes. PMID- 15781997 TI - Peroral infectivity of Toxoplasma gondii in bile and feces of interferon-gamma knockout mice. AB - Toxoplasama gondii appeared in the bile and feces of interferon-gamma knockout (GKO) but not wild type mice on days 7-8 after peroral infection with T. gondii cysts of Fukaya strain. Both tachyzoite-specific SAG1 and bradyzoite-specific T.g. HSP30 mRNAs were detected in the bile and feces of GKO mice. Tachyzoites converted to bradyzoites by culturing in the bile. By feeding uninfected mice with the bile and washed feces of T. gondii-infected GKO mice, T. gondii-specific antibody formation in the serum and cyst formation in the brain were observed. The novel migration route of T. gondii from liver to bile and feces in GKO mice was confirmed. PMID- 15781998 TI - Changes in ganglioside content affect the binding of Clostridium perfringens epsilon-toxin to detergent-resistant membranes of Madin-Darby canine kidney cells. AB - Epsilon-toxin (ET) of Clostridium perfringens, which causes fatal enterotoxemia in ungulates, was previously shown to bind to and form a heptameric pore within the detergent-resistant membranes (DRMs) of MDCK cells. Depletion of cholesterol has also been shown to decrease the cytotoxicity of ET and its heptamerization. In this study, we investigated the effects of changes in sphingolipids, other DRM components of MDCK cells, on the cells' susceptibility to ET. Treatment with fumonisin B1 and PDMP, inhibitors of sphingolipid and glycosphingolipid syntheses, respectively, increased the susceptibility, while D609, a sphingomyelin synthesis inhibitor, had the opposite effect. The exogenous addition of ganglioside G(M1) dramatically decreased the ET binding, heptamerization and cytotoxicity. These effects were shown not to be due to ET binding to G(M1) or to denaturation of ET. We also found that the ET cytotoxicity towards MDCK cells decreased with an increase in culture time. In accordance with the resistance observed for prolonged cultured cells, G(M3), a major ganglioside component, increased and sialidase treatment increased their susceptibility. These results suggest that membrane-anchored sialic acid of G(M3) within DRMs inhibits ET binding, leading to prevention of the heptamerization of ET and cell death. It is also suggested that sialidase produced by this organism aids the targeting of ET to MDCK cells. PMID- 15781996 TI - Duplex shuttle PCR for differential diagnosis of budgerigar fledgling disease and psittacine beak and feather disease. AB - Two common viral diseases in psittacine birds including budgerigar fledgling disease (BFD), generally called avian polyomavirus (APV) infection, and psittacine beak and feather disease (PBFD) have similar clinical manifestations characterized by feather disorders. A duplex shuttle PCR was developed for detection of APV and PBFD virus (PBFDV). Two pairs of oligonucleotide primers were designed to amplify a 298-bp fragment of the t/T antigen region of APV genome and a 495-bp fragment of the capsid protein region encoded by open reading frame (ORF) C1 of PBFDV genome, respectively. In the present study, APV and PBFDV were detected simultaneously in one tube by duplex shuttle PCR using these two pairs of primers. The detection limits were 2 viral copies of APV and 3 viral copies of PBFDV. In the clinical application, we detected 16 APV-positive, 15 PBFDV-positive, and 3 mixed infected samples in 39 samples examined. Sequences of the amplified products were read. The t/T antigen region was conserved in the APV positive samples as expected. ORF C1 of PBFDV genome showed diversity. Phylogenic analysis indicated that PBFDV ORF C1 consisted of 6 clusters which were related to subfamilies of psittacine birds. Our duplex shuttle PCR could be a useful method for differential diagnosis and molecular epidemiology of BFD and PBFD. PMID- 15781999 TI - Quantitative microarray-based DNA-DNA hybridization assay for measuring genetic distances among bacterial species and its application to the identification of family Enterobacteriaceae. AB - Quantitative DNA-DNA hybridization to measure the genetic distances among bacterial species is indispensable for taxonomical determination. In the current studies, we developed a method to determine bacterial DNA relatedness on a glass microarray. Reference DNAs representing a total 93 species of Enterobacteriaceae were arrayed on a glass microplate, and signal intensities were measured after 2 hr of hybridization with Cy3-labeled bacterial DNAs. All immobilized DNAs from members of the family Enterobacteriaceae were identified by this method except for DNAs from Yersinia pseudotuberculosis and Y. pestis. These results suggest that quantitative microarray hybridization could be an alternative to conventional DNA-DNA hybridization for measuring chromosome relatedness among bacterial species. PMID- 15782000 TI - Nonstructural proteins 4A and 4B of hepatitis C virus transactivate the interleukin 8 promoter. AB - Interleukin 8 (IL-8) is induced in many cell types by various stimuli including virus infection. It was reported that nonstructural protein 5A (NS5A) of hepatitis C virus (HCV) was involved in induction of IL-8 expression at both mRNA and protein levels in cultured human cells. In this study, we aimed to determine whether or not another HCV protein(s) transactivates the IL-8 gene expression, by means of an IL-8 promoter-driven luciferase reporter assay and measurement of endogenous IL-8 mRNA and secreted IL-8 protein levels. We observed that NS4B, and NS4A to a lesser extent, significantly transactivated the IL-8 promoter, which resulted in enhanced production of IL-8 protein. Also, the IL-8 expression was augmented in Huh-7 cells harboring an HCV subgenomic RNA replicon, compared with the control cells. Deletion mutational analysis of the IL-8 promoter revealed the possible involvement of the transcription factor AP-1 in both NS4A- and NS4B mediated IL-8 gene activation. In addition, the IL-8 gene activation by NS4B, but not that by NS4A, was likely to involve NF-kappaB and/or NFIL-6. The degree of the transactivation by NS4B and NS4A varied with different human cell lines, with HeLa cells showing the strongest activation followed by Huh-7 cells, and with HepG2 cells exhibiting a marginal level of activation. Taken together, our present results suggest the possibility that NS4B and NS4A play an important role in inducing the IL-8 gene expression under certain cellular conditions, which might be one of the strategies to establish persistent HCV infection. PMID- 15782001 TI - Genotyping of Norovirus strains detected in outbreaks between April 2002 and March 2003 in Osaka City, Japan. AB - Noroviruses (NVs) are the major cause of food- and waterborne nonbacterial gastroenteritis in Japan. Between April 2002 and March 2003, a total of 111 fecal specimens from 40 outbreaks of acute nonbacterial gastroenteritis in Osaka City, Japan were subject to NV detection. Seventy-two samples (64.9%) from 31 outbreaks (77.5%) were NV positive by a real time reverse transcription (RT)-PCR assay. To further determine the genotype of individual NV strains, we sequenced the capsid N-terminal/shell (N/S) domain of some representative strains from each outbreak. The 51 NV strains detected in this study were segregated into 15 genotypes (6 in genogroup I and 9 in genogroup II), and GII/5 genotype NV was a dominant outbreak genotype. PMID- 15782002 TI - Molecular basis for adaptation of a chimeric dengue type-4/Japanese encephalitis virus to Vero cells. AB - The premembrane and envelope (E) genes of a full-length cDNA clone of the dengue type-4 (DEN4) virus 814669 strain were replaced with those of the Japanese encephalitis (JE) virus JaOH0566 strain. The in vitro-synthesized RNA transcripts prepared from chimeric cDNA were used to transfect mosquito C6/36 cells. A viable chimeric virus (designated DEN4/JE) was recovered. Unexpectedly, DEN4/JE exhibited restricted growth in Vero cells. After a serial passage in Vero cells, the Vero-adapted chimeras were obtained (two clones, designated Strain I and Strain II, respectively). The entire genomes of DEN4/JE, Strain I, and Strain II were sequenced and compared. There were multiple mutations, but amino acid substitutions occurred only in E and nonstructural (NS) protein NS4B. Our findings in this study indicate that the 5' nontranslated region, E, and NS4B may be involved in Vero cell adaptation in this chimeric system. PMID- 15782003 TI - Subdural hyperintense band on diffusion-weighted imaging of chronic subdural hematoma indicates bleeding from the outer membrane. AB - The diffusion-weighted magnetic resonance (MR) imaging characteristics of chronic subdural hematoma and the correlation between hematoma liquidity and apparent diffusion coefficient (ADC) were investigated in 26 consecutive patients, 16 males and 10 females aged 42 to 92 years (mean +/- SD 73.3 +/- 13.1 years), with 31 chronic subdural hematomas. The chronic subdural hematomas were divided into homogeneous, separate, and trabecular types based on diffusion-weighted MR imaging findings. Almost all hematomas were low intensity on diffusion-weighted imaging, and the mean ADC value was 1.81 +/- 0.79 x 10(-3) mm2/sec. The high intensity areas in the subdural hematomas consisted of several types: high intensity line along the dura mater (subdural hyperintense band), high intensity along the intrahematoma septum, and laminar shape along the inner membrane. The subdural hyperintense bands accounted for almost all high intensity areas in the subdural hematomas. The mean ADC value of the high intensity areas was 0.76 +/- 0.24 x 10(-3) mm2/sec, close to that of the normal brain. The subdural hyperintense bands were considered to be intracellular and/or extracellular methemoglobin based on the T1- and T2-weighted imaging and intraoperative findings. The subdural hyperintense band is an important finding indicating relatively fresh bleeding from the outer membrane. Diffusion-weighted imaging shows liquid subdural hematoma as low intensity, and measurement of the ADC values can differentiate between liquid and solid components of the chronic subdural hematoma. PMID- 15782004 TI - Management and survival of pineoblastoma: an analysis of 34 adults from the brain tumor registry of Japan. AB - Pineoblastoma is a rare tumor in adults, and factors influencing survival are poorly understood. Data from the Brain Tumor Registry of Japan (BTRJ) was analyzed to examine patient, tumor, and treatment characteristics associated with increased survival in adults with pineoblastomas. All pineoblastoma cases in adults aged 16 years or older were identified in the BTRJ. Data were extracted on demographics, presentation, tumor characteristics, treatments, and outcomes. Kaplan-Meier plots, the log rank method, and p value <0.15 was used to screen variables for inclusion in a multivariate Cox regression estimating survival. In the final Cox multivariate model, all variables with p values <0.05 were considered significant predictors of survival, and all variables with p values 0.05-0.099 were considered trends. The BTRJ contained 34 adults with pineoblastomas diagnosed from 1969-1998. The patients were predominantly male (22 patients), with a median age of 35 years (range 16-66 years). Median survival from diagnosis was 25.7 months, with a median follow up of 20.5 months. Median surgical resection was 75-94%, and five of the 34 patients had gross total resection. Twenty-nine of the 34 patients received cranial irradiation therapy with a median dose of 50 Gy (range 30-70 Gy). In the final multivariate model, cranial irradiation > or =40 Gy (p=0.014) and gross total resection (p=0.034) were associated with improved survival. There was a trend towards improved survival for women (p=0.099). Adult pineoblastoma patients have poor survival prognosis. Cranial irradiation therapy using at least 40 Gy and complete surgical resection are associated with improved survival. PMID- 15782005 TI - Incomplete oculomotor nerve palsy caused by an unruptured internal carotid anterior choroidal artery aneurysm--case report--. AB - A 59-year-old woman visited our institute with the chief complaint of dizziness which persisted whenever she tried to focus on objects. She had not experienced apparent double vision and had no history of intracranial bleeding. Neurological examination revealed no abnormality except for exotropia at the mid-position and at upper gaze. Cerebral angiography revealed that the intracranial portion of the left internal carotid artery ran more horizontally and also identified an unruptured left internal carotid-anterior choroidal artery (IC-AChA) aneurysm of 3.0 mm diameter. The aneurysm at the origin of the AChA was confirmed during surgery. The proximal lateral wall of the aneurysm was in contact with the oculomotor nerve. This contact was released after complete obliteration of the aneurysm. The exotropia resolved 3 months later. Oculomotor nerve palsy usually indicates the presence of internal carotid-posterior communicating artery (IC PcomA) aneurysm. Since sacrifice of the AChA will result in severe neurological deficits, accurate neuroimaging information is needed prior to the operation. Conventional angiography and/or three-dimensional computed tomography angiography should be performed to ascertain whether the aneurysm is an IC-PcomA or IC-AChA aneurysm, even if some neurosurgeons insist that conventional angiography is not always needed before surgery for an unruptured aneurysm. PMID- 15782006 TI - Spontaneous intracranial internal carotid artery dissection treated by intra arterial thrombolysis and superficial temporal artery-middle cerebral artery anastomosis in the acute stage--case report--. AB - A 22-year-old man presented with sudden onset of right retro-orbital headache followed by left hemiparesis. Right carotid angiography demonstrated almost total occlusion of the intracranial internal carotid artery (ICA) and severe stenosis of the middle cerebral artery (MCA), presumably caused by arterial dissection. Local arterial injection of urokinase was performed 2 hours after onset. The ICA became patent, but the M2 portion of the MCA was still occluded, and the left hemiparesis did not improve. Superficial temporal artery-MCA anastomosis was immediately performed. The left hemiparesis disappeared completely 6 days after this procedure. Angiography 2 weeks after the onset revealed occlusion of the ICA, and maintenance of blood flow to the right cerebral hemisphere via the anastomosis. Magnetic resonance imaging showed small infarcts in the right cerebral cortex. Repeat angiography after 5 months showed recanalization of the right ICA and the right MCA. Combination of thrombolytic therapy and bypass surgery may be a useful treatment option for patients with sudden occlusion of the intracranial artery caused by dissection. PMID- 15782007 TI - Three-dimensional computed tomography angiography for the investigation of superficial temporal artery pseudoaneurysms--two case reports--. AB - An 89-year-old man and a 60-year-old man presented with superficial temporal artery (STA) pseudoaneurysms which developed secondary to trauma. Conventional cerebral angiography and three-dimensional computed tomography (3D CT) angiography clearly demonstrated the STA pseudoaneurysms. The patients underwent surgical excision of the aneurysms based on the conventional cerebral angiography findings in one patient and 3D CT angiography findings in other patient. 3D CT angiography is an excellent noninvasive diagnostic method for detecting extracranial aneurysms such as STA pseudoaneurysm, especially the relationship between the aneurysm and surrounding structures, including the calvarium. PMID- 15782008 TI - Primary cerebral angiitis containing marked xanthoma cells with massive intraparenchymal involvement--case report--. AB - A 27-year-old woman was referred to our hospital with mild disorientation, bilateral abducens nerve palsy, and mild left hemiparesis. Magnetic resonance (MR) imaging revealed diffuse mass lesions resembling malignant glioma in the right frontal intraparenchymal region, with enhancement of multiple meningeal and intraparenchymal nodules. Partial resection of the frontal lesion was performed. Histological examination revealed that the specimens consisted of brain tissue, with marked perivascular infiltration of histiocytes and sheets of xanthomatous cells. The diagnosis was primary cerebral angiitis containing marked xanthoma cells. Steroid therapy was administered over 1 week. MR imaging showed that the remaining lesions resolved gradually, and had disappeared 2 years after surgery. No neurological symptoms or recurrence of the tumor has been observed during the 6-year period since the operation. PMID- 15782009 TI - Stereotactic radiosurgery for recurrent pleomorphic adenoma invading the skull base--case report--. AB - A 38-year-old man presented with a recurrent pleomorphic adenoma in the parapharyngeal space invading the skull base 19 years after the first operation for a parotid gland tumor. Stereotactic radiotherapy was performed to control the tumor growth using a marginal dose of 8 Gy and maximum dose of 18 Gy with care taken to minimize the dose to nearby structures. The symptoms were reduced within a few months. Magnetic resonance imaging over 5 years showed that the tumor was controlled with no regrowth. Stereotactic radiotherapy is a therapeutic option for the treatment of pleomorphic adenomas. PMID- 15782010 TI - Giant intrasellar arachnoid cyst manifesting as adrenal insufficiency due to hypothalamic dysfunction--case report--. AB - A 67-year-old man first noticed loss of pubic and axillary hair in 1992 and then a visual field defect in 2001. He experienced loss of consciousness attributed to hyponatremia in April 2002. Magnetic resonance imaging showed a giant intrasellar cystic mass, 40 mm in diameter, that had compressed the optic chiasm. The patient complained of chronic headache, and neurological examination revealed bitemporal hemianopsia. Preoperative endocrinological examination indicated adrenal insufficiency, and hypothyroidism due to hypothalamic dysfunction. The patient underwent endonasal transsphenoidal surgery. The cyst membrane was opened and serous fluid was drained. Histological examination identified the excised cyst membrane as arachnoid membrane. The patient's headaches resolved postoperatively, but the bitemporal hemianopsia and endocrinological function were unchanged. This arachnoid cyst associated with hypothalamic dysfunction might have been caused by an inflammatory episode in the suprasellar region. PMID- 15782011 TI - Spinal tanycytic ependymoma with hematomyelia--case report--. AB - A 58-year-old man presented with an extremely rare case of "pure type" spinal tanycytic ependymoma associated with hematomyelia manifesting as sensory disturbance of the bilateral hands and weakness of the right arm. Magnetic resonance imaging demonstrated a tumor in the spinal cord from C-2 to C-4 levels. The soft gelatinous tumor was subtotally resected and the adjacent chronic liquid hematoma was aspirated. The immunohistochemical and ultrastructural findings indicated a diagnosis of tanycytic ependymoma. PMID- 15782012 TI - Knife blade penetrating stab wound to the brain--case report--. AB - A 28-year-old man attempted to kill himself with a knife stab into the parietal area. Neuroimaging showed no vascular impairment except slow venous flow around the knife due to tamponading. After obtaining informed consent, the knife was removed through a craniotomy without new brain injury. Postoperative neurological findings showed no deficit. Follow-up angiography revealed no vascular impairment. No infection occurred. Brain stab wounds cause numerous complications, such as intracranial hemorrhage, injury of important vessels, and infections. Minimal blade movement during removal and precautions to prevent massive hemorrhage are essential. PMID- 15782013 TI - Outcome of endovascular treatment in symptomatic intracranial vascular stenosis. AB - OBJECTIVE: The outcome evaluation for the revascularization of intracranial vascular stenoses has not been fully described due to the highly technical nature of the procedure. We report here on the early and late clinical outcomes of angioplasty and/or stenting of symptomatic severe intracranial vascular stenoses at a single institute. MATERIALS AND METHODS: Since 1995, we have treated 35 patients with symptomatic intracranial vascular stenosis (more than 70% stenosis, mean stenosis: 78.6%+/-6.2%). Angioplasty (n = 19) was performed for the horizontal segment of the middle cerebral artery (M1) (n = 16) and the basilar artery (BA) (n = 1), the intradural vertebral artery (VA) (n = 1), and the cavernous internal carotid artery (ICA) (n = 1). Stenting (n = 16) was performed for the cavernous or petrous ICAs (n = 9), the intradural VA (n = 3), BA (n = 2), and M1 (n = 2) artery. We assessed the angiographic success (defined as residual stenosis < 50%) rate, the periprocedural complications during the 30-day periprocedural period, the symptomatic recurrence and restenosis during a mean 22 month follow-up (FU) period. The Kaplan-Meier estimate of the cumulative event free rate of the major cerebrovascular events, i.e. death, stroke or restenosis, was also done. RESULTS: Angiographic success was achieved in 97% of our patients (34/35). There were four procedure-related complications (11%) including a death and a minor stroke. During the mean 22-month FU, the asymptomatic restenosis rate was 9% and the symptomatic restenosis rate was 6% in the target lesion and 9% in all the vascular territories. The Kaplan-Meier estimate was 70.6% (95% confidence interval = 46.5-94.7) after 33 months of FU. CONCLUSION: In addition to a high angiographic success rate and an acceptable periprocedural complication rate, intracranial angioplasty and/or stenting revealed a relatively low symptomatic recurrence rate. Hemorrhage is a rare, but the physician must aware that potentially fatal periprocedural complications can occur. PMID- 15782015 TI - Detectability of various sizes of honeycombing cysts in an inflated and fixed lung specimen: the effect of CT section thickness. AB - OBJECTIVE: We wanted to clarify the relationship between the visibility of air cysts on CT images, the CT slice thickness and the size of the air cysts, with contact radiographs as the gold standard, for the accurate evaluation of honeycomb cysts. MATERIALS AND METHODS: An inflated and fixed autopsied lung having idiopathic interstitial pneumonia was evaluated. The corresponding air cysts were identified on the contact radiographs of a 0.5 mm-thick-section specimen and also on the CT images of three different kinds of section thickness: 0.5, 1.0 and 2.5 mm. The maximal diameters of the air cysts were measured under a stereomicroscope. RESULTS: A total of 341 air cysts were identified on the contact radiograph, and they were then evaluated. Sixty-six percent of air cysts 1 to 2 mm in diameter were detected by 0.5 mm slice thickness CT, while only 34% and 8% were detected by 1.0 and 2.5 mm slice thickness CT, respectively. Only 28% and 22% of air cysts less than 1 mm in diameter were detected by 0.5 and 1.0 mm slice thickness CT, respectively. CT with a 2.5 mm slice thickness could not demonstrate air cysts less than 1 mm in diameter. CONCLUSION: The CT detection rate of honeycombing is significantly influenced both by the slice thickness and the size of the air cysts. PMID- 15782014 TI - Assessment of the prognostic factors for a local recurrence of rectal cancer: the utility of preoperative MR imaging. AB - OBJECTIVE: To determine the utility of MR imaging in evaluating the prognostic factors for a local recurrence of rectal cancer following a curative resection. MATERIALS AND METHODS: The preoperative MR images obtained from 17 patients with a local recurrence and 54 patients without a local recurrence, who had undergone a curative resection, were independently evaluated by three radiologists. The following findings were analyzed: the direct invasion of the perirectal fat by the primary rectal carcinoma, involvement of the perirectal lymph nodes, perirectal spiculate nodules, perivascular encasement, and an enlargement of the pelvic wall lymph nodes. The clinical and surgical profiles were obtained from the patients' medical records. The association of a local recurrence with the MR findings and the clinicosurgical variables was statistically evaluated. RESULTS: Of the MR findings, the presence of perivascular encasement (p = 0.001) and perirectal spiculate nodules (p = 0.001) were found to be significant prognostic factors for a local recurrence. Of the clinicosurgical profiles, the presence of a microscopic vascular invasion (p = 0.005) and the involvement of the regional lymph nodes (p = 0.006) were associated with a local recurrence. Logistic regression analysis showed that the presence of perirectal spiculate nodules was an independent predictor of a local recurrence (odds ratio, 7.382; 95% confidence interval, 1.438, 37.889; p = 0.017). CONCLUSION: The presence of perirectal spiculate nodules and perivascular encasement on the preoperative MR images are significant predictors of a local recurrence after curative surgery for a rectal carcinoma. This suggests that preoperative MR imaging can provide useful information to help in the planning of preoperative adjuvant therapy. PMID- 15782016 TI - CT and MR imaging of the buccal space: normal anatomy and abnormalities. AB - The buccal space is an anatomical compartment lying anterior to the masticator space and lateral to the buccinator muscle. Since the major purpose of imaging is to define the likely anatomic origin and also the extent of a given lesion, thorough knowledge of the normal anatomy of the buccal space is essential, and this knowledge can aid the physician in narrowing down the list of possible maladies on the differential diagnosis. We illustrate here in this paper the important anatomic landmarks and typical pathologic conditions of the buccal space such as the developmental lesions and the neoplastic lesions. Knowledge of the expected pathologic conditions is useful for the radiologist when interpreting facial CT and MR images. PMID- 15782018 TI - Posttraumatic pseudoaneurysm in scalp treated by direct puncture embolization using N-butyl-2-cyanoacrylate: a case report. AB - Here, we report a case of scalp pseudoaneurysm which was treated by direct puncture embolization using n-butyl-2-cyanoacrylate. The patient had a history of blunt trauma in the previous two months. Ultrasound-guided manual compression was initially attempted, but the results were unsatisfactory. Direct puncture embolization was then performed, and the pseudoaneurysm was completely obliterated. Non-surgical treatment options for pseudoaneurysm are briefly discussed. PMID- 15782017 TI - Ductographic findings of breast cancer. AB - Ductography has become the gold standard for the evaluation of patients exhibiting pathologic nipple discharges. In nine patients (age range, 29-67 years; median age, 51 years) with invasive (n=5) or intraductal (n=4) cancer, ductographic findings were recorded, then correlated with mammographic and sonographic findings. Common ductographic findings included complete ductal obstruction, multiple irregular filling defects in the nondilated peripheral ducts, ductal wall irregularities, periductal contrast extravasation, and ductal displacement. Faint microcalcifications or ill-defined masses, which were not opacified by contrast material, were often discovered adjacent to ductal abnormalities. Mammographically and sonographically occult diffusely spreading intraductal cancers often manifested as pathologic nipple discharge. In such cases, meticulous ductographic examinations and interpretations were crucial in order not to miss breast cancers. PMID- 15782019 TI - Bilateral paramedian thalamic contrast enhancement on CT after Intra-arterial thrombolysis. AB - A 58-year-old woman presented with an acute embolic occlusion of the distal basilar artery. She underwent angioplasty and intra-arterial thrombolysis. Angiography performed after recanalization revealed a single perforating thalamic artery. A nonenhanced CT scan carried out immediately after the procedure revealed hyperdense lesions in the bilateral paramedian portions of the thalami, which disappeared on the 24-hour follow-up CT scan. Three months later, the patient improved to functional independence, but had some memory dysfunction and vertical gaze palsy. This case suggests that contrast enhancement or extravasation can occur in the thalamus after intra-arterial thrombolysis performed to recanalize a basilar artery occlusion. PMID- 15782020 TI - MR imaging of pregnancy luteoma: a case report and correlation with the clinical features. AB - We report here on a 26-year-old pregnant female who developed hirsutism and virilization during her third trimester along with a significantly elevated serum testosterone level. Abdominal US and MR imaging studies were performed, and they showed unique imaging features that may suggest the diagnosis of pregnancy luteoma in the clinical context. After the delivery, the serum testosterone level continued to decrease, and it returned to normal three weeks postpartum. The follow-up imaging findings were closely correlated with the clinical presentation. PMID- 15782021 TI - Fatigue stress fractures of the pubic ramus in the army: imaging features with radiographic, scintigraphic and MR imaging findings. AB - Although fatigue fractures are not unusual in athletes and military personnel those of the pubic ramus are rare. We report three cases of fatigue fractures of the inferior pubic rami in two male recruits and one female military cadet. On the initial radiograph, most of the lesions were subtle and easy to overlook. However, bone scintigraphy provided more distinct images that allowed easy and early detection of lesions, and MR imaging presented more diagnostic information, which allowed a precise diagnosis. PMID- 15782022 TI - Bilateral transverse (Bowdler) fibular spurs with hypophosphatasia in an adolescent girl. AB - Hypophosphatasia is a clinically heterogeneous inheritable disorder characterized by defective bone mineralization and the deficiency of serum and tissue liver/bone/kidney alkaline phosphatase activities. Due to the mineralization defect of the bones, various skeletal findings can be radiologically observed in hypophosphatasia. Bowing and Bowdler spurs of long bones are the characteristic findings. The Bowdler spurs reported on in the previous pertinent literature were observed in the perinatal aged patients and these lesions have rarely involved adolescents. We herein report on a 14-year-old girl with fibular Bowdler spurs. PMID- 15782023 TI - Safety, scrutiny and conflicts: assessing the fallout and lessons learned from pharmaceuticals. PMID- 15782024 TI - Low-energy laser irradiation promotes cellular redox activity. AB - Low-energy visible light (LEVL) has been shown to stimulate cell functions. This is called "photobiostimulation" and has been used successfully over the last three decades for treating a range of conditions, including soft tissue injuries, severe wounds, chronic pain, and more. Nevertheless, the mechanism of photobiostimulative processes is still being debated. It is obvious that, in order to interact with the living cell, light has to be absorbed by intracellular chromophores. In a search for chromophores responsible for photobiostimulation, endogenous porphyrins, mitochondrial and membranal cytochromes, and flavoproteins were found to be suitable candidates. The above-mentioned chromophores are photosensitizers that generate reactive oxygen species (ROS) following irradiation. As the cellular redox state has a key role in maintaining the viability of the cell, changes in ROS may play a significant role in cell activation. In the present review, we summarize evidence demonstrating that various ROS and antioxidants are produced following LEVL illumination. We found that very little evidence for NO formation in illuminated non-vascular smooth muscle cells exists in the literature. We suggest that the change in the cellular redox state which plays a pivotal role in maintaining cellular activities leads to photobiostimulative processes. PMID- 15782025 TI - Pulpal and periodontal temperature rise during KTP laser use as a root planing complement in vitro. AB - OBJECTIVE: The purpose of this study was to define the optimal irradiation conditions of a KTP laser during root planing treatment. METHODS: The surfaces of 60 single-root human teeth were scaled with conventional instruments before lasing. The pulpal temperature increase was measured by means of one thermocouple placed in the pulp chamber and a second one placed on the root surface at 1 mm from the irradiation site. The influence of variables of coloration by Acid Red 52 (photosensitizer), scanning speed, dentin thickness, and probe position was analyzed for a constant exposure time of 15 sec and 500 mw (spot size diameter, 0.5 mm). The pulpal temperature was below 3 degrees C for the adjustments. RESULTS: The irradiation on one point of root surface had the following results: The application of photosensitizer on the root surface before lasing produced a 50% higher temperature rise within the pulp than in the case without the application of the photosensitizer. The temperature rise in the pulp chamber was below 3 degrees C with the following settings of 500 mw: PW = 10 msec and PRR < 35; or PW= 20 msec and PRR < 20 Hz. On the other hand, for the same irradiation conditions, the temperature rise on the surface of the root was always below 7 degrees C. However, the temperature increase became higher than 7 degrees C (on the surface of the root) in the case of P > 500 mw, PW > 50 msec and PRR > 10 Hz of root surface or a scanning speed of irradiation of 1 mm/sec for a linear irradiation of 4 mm. CONCLUSION: The KTP laser may be used safely without thermal damage to pulp and periodontal tissue with respect to the biologically acceptable previously described parameters. PMID- 15782026 TI - 940-nm laser for treatment of saphenous insufficiency: histological analysis and long-term follow-up. AB - OBJECTIVE: Currently, many methods are available for the treatment of saphenous insufficiency. Modalities available include surgery, sclerotherapy, and radiofrequency closure. Recently, a method has been introduced using laser energy for the treatment of the saphenous vein using a bare-tipped fiber. Our experience using the 940-nm wavelength forms the basis of this report. METHODS: Indications for treatment with laser obliteration alone was saphenous insufficiency, a saphenofemoral junction of less than 15 mm in diameter and absence of concurrent anticoagulation therapy. Ninety-two percent of the patients were in Class C-2 to C-4, and 8% were in C-5 to C-6. Eight percent had a concurrent high ligation of the saphenous vein. Fifteen patients were treated at varying energy levels and time durations for saphenous insufficiency using a bare-tipped fiber and the 940 nm wavelength laser (SkinPulse S by Dornier). These patients then had histologic evaluation of the treated vein. An additional 620 patients were subsequently treated for saphenous insufficiency using the 940- nm wavelength laser. RESULTS: At 1-sec pulse durations, histologic studies reveal endothelium and subendothelial damage as the initial response to laser damage. At 3 months, endothelium was still absent, with organized thrombus in the lumen. At 6 months, at 1-sec pulse duration, thrombus organization was still present with evidence of muscle wall damage. Ultrasound evaluation revealed gradual diminution in saphenous vein diameter over 6 months, with absence of reflux and normal flow at the saphenofemoral junction in 95% of patients. For pulse durations of greater than 1 sec, there was transmural injury. CONCLUSION: Laser treatment of the saphenous vein was associated with no complications in our series of patients. The vein remained closed in 95% of those treated. This technique may replace other treatment modalities in up to 90% of patients. PMID- 15782027 TI - Healing process of skin after CO2 laser ablation at low irradiance: a comparison of continuous-wave and pulsed mode. AB - OBJECTIVE: The purpose of the present study was to compare the healing process of rabbit's skin after the ablation of continuous wave CO(2) laser and pulsed CO(2) laser at low irradiances. BACKGROUND DATA: Because of its advantages, the CO(2) laser has been viewed as an alternative to the traditional scalpel. Simple thermal models suggest that suitably short pulses of laser irradiation can leave a smaller thermal damaged zone to ablated tissue, which may lead to faster healing. METHODS: One continuous-wave (cw, 2 watt) and two pulsed CO(2) lasers (sp(1), 100 Hz, 2 watt; and sp(2), 25 Hz, 2 watt) were used to ablate twelve rabbits' ear skin for 2 min, respectively. After sacrifice at 0, 3, 7, 14, 28, and 56 days postsurgery, gross observation and histological examination were performed. RESULTS: sp(1) and sp(2) resulted in similar ablation rates to cw laser. At 3, 7, 14, and 28 days postsurgery, the scores of the group cw were significantly lower than both the sp(1) and sp(2) scores (p < 0.01). From 7 to 28 days, wounds of sp1 showed the best histological outcomes. CONCLUSION: At a relatively low irradiance, pulsed CO(2) laser is capable of bloodless skin ablation with improved wound healing. Increasing repetition rate of the CO(2) laser may help to achieve a better surgical outcome. Pulsed CO(2) laser may be a valuable instrument for ablation skin and skin lesions. PMID- 15782028 TI - Infrared laser light reduces loading time of dental implants: a Raman spectroscopic study. AB - OBJECTIVE: The aim of this study was to assess, through near-infrared Raman spectroscopy (NIRS), the incorporation of hydroxyapatite of calcium (CHA; approximately 960 cm(1))--on the healing bone around dental implants submitted or not to low-level laser therapy (LLLT) (lambda830 nm). BACKGROUND DATA: The process of maturation of the bone is important for the success of dental implants, as it improves the fixation of the implant to the bone, allowing the wearing of a prosthesis. LLLT has been suggested as a mean of improving bone healing because of its biomodulatory capabilities. METHODS: Fourteen rabbits received a titanium implant on the tibia; eight of them were irradiated with lambda830-nm laser (seven sessions at 48-h intervals, 21.5 J/cm(2) per session, 10 mW, phi approximately 0.0028 cm(2), 85 J/cm(2) treatment dose), and six acted as control. The animals were sacrificed at 15, 30, and 45 days after surgery. Specimens were routinely prepared for Raman spectroscopy. Twelve readings were taken on the bone around the implant. RESULTS: The results showed significant differences in the concentration of CHA on irradiated and control specimens at both 30 and 45 days after surgery (p < 0.001). CONCLUSION: It is concluded that LLLT does improve bone healing, and this can be safely assessed by Raman spectroscopy. PMID- 15782029 TI - Nociceptive scores and endorphin-containing cells reduced by low-level laser therapy (LLLT) in inflamed paws of Wistar rat. AB - OBJECTIVE: This study aimed to investigate how local pain relief is mediated by laser therapy and how dose affects the relationship. METHODS: Inflammation was induced in the hind-paws of Wistar rats. Two groups of rats received 780-nm laser therapy (Spectra-Medics Pty Ltd.) at one of two doses (2.5 and 1 J/cm(2)). One group acted as a control. Scores of nociceptive threshold were recorded using paw pressure and paw thermal threshold measures. RESULTS: A dose of 1 J/cm(2) had no statistically significant effect on antinociceptive responses. A dose of 2.5 J/cm(2) demonstrated a statistically significant effect on paw pressure threshold ( p < 0.029) compared to controls. There was no difference in paw thermal threshold responses and paw volumes at either dose. Immunohistochemistry in control animals demonstrated normal beta-endorphin containing lymphocytes in control inflamed paws but no beta-endorphin containing lymphocytes in rats that received laser at 2.5 J/cm(2). CONCLUSION: The results confirm previous findings that the effect of laser therapy is dose-related. The mechanism of effect may occur via a differentiated pressure-sensitive neural pathway rather than a thermal-sensitive neural pathway. The significance of the immunohistochemistry findings remains unknown. PMID- 15782030 TI - Evaluation of retention of post-core system cemented with different materials on dentine surfaces treated with EDTA or Er:YAG laser irradiation. AB - OBJECTIVE: The aim of this study was to evaluate in vitro the post-core system retention to dentine surfaces treated with EDTA and Er:YAG laser irradiation and cemented with resin or a tradiational zinc phosphate cement. BACKGROUND DATA: One of the factors related to intracanal post-retention is the root canal wall preparation. METHODS: Forty-eight samples were divided into three groups: (1) G1 (control), dentine surfaces with no treatment, irrigated with distilled and deionized water; (2) G2, 1% NaCIO + 17% EDTA; and (3) G3, water and Er:YAG laser (8 Hz, 200 mJ, 60 J, 300 pulses). Each group was subdivided into two subgroups depending on the cement used: Panavia F or zinc phosphate. All specimens were submitted to tensile load in the Universal Instron Machine (Instron 4444) at 0.5 mm/min. RESULTS: Data were analyzed statistically with ANOVA and Tukey's test, which showed no significant difference (p > 0.01) between the dentine wall treatments performed with 1% NaCIO + 17% EDTA (6.80 Mpa) and water + Er:YAG laser (6.81 MPa), independent of the cement used; however, these were statistically different (p < 0.01) from the control group that presented the lowest values (4.82 MPa). Cementing with Panavia F and zinc phosphate presented similar results for adhesive resistance (p > 0.05). CONCLUSION: We conclude that the previous treatment of dentine walls with 1% NaCIO + 17% EDTA, or Er:YAG laser + water favored retention of the post-core system cemented with both resin and zinc phosphate cement. PMID- 15782031 TI - Temperature analysis during bonding of brackets using LED or halogen light base units. AB - The purpose of our investigation is to compare the intrapulpal temperature changes following blue LED system and halogen lamp irradiation at the enamel surface of permanent teeth. The fixation of brackets using composite resin is more comfortable and faster when using a photo-curable composite. Several light sources can be used: halogens, arc plasma, lasers, and recently blue LED systems. An important aspect to be observed during such a procedures is the temperature change. In this study, we have used nine human extracted permanent teeth: three central incisors, three lateral incisors, and three canines. Teeth were exposed to two light sources: blue LED system (preliminary commercial model LEC 470-II) and halogen lamp (conventional photo-cure equipment). The surface of teeth was exposed for 20, 40, and 60 sec at the buccal and lingual enamel surface with an angle of 45 degrees. Temperature values measured by a thermistor placed at pulpar chamber were read in time intervals of 1 sec. We obtained plots showing the temperature evolution as a function of time for each experiment. There is a correlation between heating quantity and exposition time of light source: with increasing exposition time, heating increases into the pulpal chamber. The halogen lamp showed higher heating than the LED system, which showed a shorter time of cooling than halogen lamp. The blue LED system seems like the indicated light source for photo-cure of composite resin during the bonding of brackets. The fixation of brackets using composite resin is more comfortable and faster when using a photo-curable composite. Blue LED equipment did not heat during its use. This could permit a shorter clinical time of operation and better performance. PMID- 15782032 TI - Experimental study on the morphological and functional recovery following partial glossectomy in rabbits: a comparison between CO2 laser and electrocautery. AB - OBJECTIVE: The aim of the present study was to evaluate and compare the morphological and functional recovery following partial glossectomy in rabbits with the CO(2) laser and electrocautery. METHODS: Partial glossectomy with the CO(2) laser or electrocautery in rabbits was performed to investigate changes in the body weight and tongue width after surgery. The study models were classified into three groups. In group 1, the excised edges of the wound were left unsutured after partial glossectomy with the CO(2) laser. In group 2, the wound was closed after CO(2) laser surgery. In group 3, as a control group, the wound was closed after partial glossectomy with electrocautery. The specimens of the tongues obtained from the rabbit were microscopically assessed. RESULTS: No significant differences were found in the percent change in the body weight among the three groups. There was a significant difference in the tongue width between groups 1 and 3 at each time point: 2 weeks (p < 0.05), 4 weeks (p < 0.01), and 8 weeks (p < 0.001) postoperatively. Furthermore, there was a significant difference in the tongue width between groups 2 and 3 at 8 weeks (p < 0.02) postoperatively. Histologically, the scar tissue of the wound was extensive in the control group, although it was localized in the laser group. CONCLUSION: Postoperative dysfunction was reduced when excised edges were left unsutured after partial glossectomy with the CO(2) laser. PMID- 15782033 TI - Morphological comparative study on ablation of dental hard tissues at cavity preparation by Er:YAG and Er,Cr:YSGG lasers. AB - OBJECTIVE: The purpose of this study was to evaluate morphologically the dental hard tissue ablation at the class V cavity preparation by two types of laser devices: Er:YAG and Er,Cr:YSGG lasers in vitro. BACKGROUND DATA: There have been no reports on the comparative study of dental tissue ablation at cavity preparation by Er:YAG and Er,Cr:YSGG lasers. METHODS: Twenty-two extracted human mature molar teeth were used in this study and divided into two groups of 11 teeth each. The teeth of the Er:YAG laser-irradiated group were irradiated at the parameters of 250 mJ/pulse and 15 Hz with water spray, and those of the Er,Cr:YSGG laserirradiated group were irradiated at the parameters of 5 Wand 20 Hz with water spray. After cavity preparation, the teeth were dehydrated, coated with platinum, and examined by scanning electron microscopy. RESULTS: Both of the laser devices were capable of ablating dental hard tissues, but similar, irregular, and rugged surface aspects with different depths were observed. Open dentinal tubules at the cavities prepared by Er,Cr:YSGG laser were more clearly visible than those prepared by Er:YAG laser. Smaller width and stripped surfaces were observed on the cavities prepared by Er,Cr:YSGG laser. CONCLUSION: The results suggested that there is little difference between the two types of lasers Er:YAG and Er,Cr:YSGG lasers-morphologically for class V cavity preparation, because both lasers were capable of preparing class V cavities, and the morphological features of the irradiated surfaces were very similar. PMID- 15782034 TI - Effects of pulsed Nd:YAG laser on microleakage of composite restorations in class V cavities. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of pulsed Nd:YAG laser on decrease of dentinal microleakage beneath the restoration. METHODS: Thirty-two sound extracted premolar teeth were subjected to two standardized, V shaped, class V cavity preparations on both buccal and lingual surfaces. Cavities were located at 1-2 mm below the cemento-enamel junction (CEJ) line. The teeth were divided into two groups of experimental and control in an random manner. Both 20 and 15 mJ/pulse laser energies were tested. Etching, bonding, and filling were performed using Tetric ceram composite material. All samples were thermocycled for 1000 cycles, followed by an immersing period of 1 week in 2% basic fuchsin solution. Specimens were then sliced longitudinally and evaluated under a digital scanner for the extent of dye penetration. RESULTS: Our results indicated that the level of microleakage of laser-treated cavities was significantly less than that of untreated cavities (p < 0.05) for 20 mJ/pulse laser energy. No significant decrease in microleakage level was observed for the 15 mJ/pulse laser energy. CONCLUSION: The short pulsed Nd:YAG laser with the energy of 20 mJ/pulse and aforementioned features appeared to be able to reduce microleakage toward pulp. PMID- 15782035 TI - Retrospective study of adjunctive diode laser therapy for pain attenuation in 662 patients: detailed analysis by questionnaire. AB - OBJECTIVE: The aim of this study was to assess the long-term effects of low-level laser therapy (LLLT) through a retrospective survey using questionnaires. BACKGROUND DATA: The use of LLLT for chronic pain attenuation has been reported in the international literature for over 20 years. METHODS: We used a series of diode laser systems in which the most effective wavelength was consistently found to be 830 nm with an output power in continuous-wave of 60 mW. Subjects were 1,087 patients treated by LLLT at the Shiroto Clinic from April 1992 to August 1995. Questionnaires were sent to subjects in September and October 1996. RESULTS: The reply rate was 60.9%, comprising 662 questionnaires (265 males, 397 females, mean age of 53.4 years). The total efficacy rating (excellent plus good) immediately after LLLT was 46.8% in men and 47% in women. At the time of the survey, this rose to 73.3% in men and 76.8% in women, with positive effects also recorded on psychosomatic factors such as well-being, physical energy, general fatigue, mental vigor, and emotional stability. LLLT effects continued for 1-3 days. No statistically significant difference in efficacy was seen between males and females. LLLT as used in the study is therefore considered safe, effective, and side-effect-free, making it an ideal adjunctive therapeutic modality for intractable chronic and other pain. CONCLUSION: Infrared diode LLLT is therefore considered safe, effective, and side-effect-free, making it an ideal adjunctive therapeutic modality for intractable chronic pain. PMID- 15782036 TI - The use of polarized polychromatic non-coherent light as therapy for acute tennis elbow/lateral epicondylalgia: a pilot study. AB - OBJECTIVE: The aim of this study was to assess the efficacy of polarized, polychromatic, non-coherent, low energy light (Bioptron 2, Bioptron AG, Switzerland) in the treatment of acute tennis elbow. BACKGROUND: Tennis elbow, or lateral epicondylitis, is one of the most common lesions affecting the arm. A plethora of treatment regimes have been described for this condition, but no specific therapy has emerged as a gold standard. METHODS: A pilot study was carried out with 25 patients who had acute tennis elbow. Bioptron 2 device was applied over lateral epicondyle three times per week for 4 weeks. Pain on VAS, function on VAS, and painfree grip strength were measured at the beginning (week 0) and at the end of the study (week 4). RESULTS: The pain on VAS was reduced at the end of treatment (t(24) = 3.84, p = 0.001). Function on VAS was increased at the end of treatment (t(24) = 4.23, p < 0.001). Pain-free grip strength was increased at the end of treatment (t(24) = 4.23, p < 0.004). CONCLUSION: Although the results suggested that the Bioptron 2 could reduce patients' symptoms with acute tennis elbow, future controlled studies are needed to establish the relative and absolute effectiveness of Bioptron 2. PMID- 15782037 TI - Low-level laser therapy (LLLT) efficacy in post-operative wounds. AB - OBJECTIVE: The aim of this paper was to investigate the efficacy of low-level laser radiation (LLLR) with wavelength of 904 nm on the stimulation of the healing process of postoperative aseptic wounds (early scar). BACKGROUND DATA: Low-level laser therapy (LLLT) has been increasingly used to treat many disorders, including wounds. However, despite such increased clinical usage, there is still controversy regarding the efficacy of this wound treatment in curent clinical practice. METHODS: LLLT has been used to treat cutting plague in the right instep and on the left foot. Both resulted from sutured wounds. The clinical evaluation by semiquantitative methods is presented. RESULTS: Clinical evaluation showed that the healing process of these postoperatively treated wounds has occurred and that the functional recovery of the patients (i.e., return to their ordinary life) was faster than without treatment. CONCLUSION: LLLR with wavelength of 904 nm to stimulate postoperative aseptic wounds (early scar) is efficient in both cases of cutting plague. PMID- 15782038 TI - Six-degrees-of-freedom sensing based on pictures taken by single camera. AB - Two six-degrees-of-freedom sensing methods are presented. In the first method, three laser beams are employed to set up Descartes' frame on a rigid body and a screen is adopted to form diffuse spots. In the second method, two superimposed grid screens and two laser beams are used. A CCD camera is used to take photographs in both methods. Both approaches provide a simple and error-free method to record the positions and the attitudes of a rigid body in motion continuously. PMID- 15782039 TI - Novel approach to fluoroscopic visualization of the lower cervical discs. PMID- 15782040 TI - Laser (and LED) therapy is phototherapy. PMID- 15782041 TI - Laser literature watch. PMID- 15782044 TI - Male athletes and eating disorders. PMID- 15782046 TI - Summary and agreement statement of the 2nd International Conference on Concussion in Sport, Prague 2004. PMID- 15782047 TI - Free gracilis tendon graft in neglected tears of the achilles tendon. AB - OBJECTIVE: To report the results of a longitudinal study on reconstruction of neglected Achilles tendon rupture using a free autologous gracilis tendon graft. DESIGN: Cohort study. PARTICIPANTS: Twenty-one patients underwent surgery for a neglected rupture of the Achilles tendon occurring between 65 days and 9 months before the operation. METHODS: All participants were prospectively followed up for 2 years, and final review was performed at 28.4 +/- 3.5 months from the operation. Functional (anthropometric measurements, isometric strength, return to activities) and clinical assessment was performed. RESULTS: No patients experienced any problems in the wound used to harvest the tendon of gracilis. Five patients were managed conservatively following a superficial infection of the Achilles tendon surgical wound. No patients developed a deep vein thrombosis or sustained a rerupture. All patients were able to walk on tiptoes, and no patient used a heel lift or walked with a visible limp. The maximum calf circumference remained significantly decreased in the operated leg at final review. The operated limb was significantly less strong than the nonoperated one. CONCLUSIONS: The management of neglected tears of the Achilles tendon by free gracilis tendon grafting is safe but technically demanding. It affords good recovery, even in patients with a neglected rupture of a duration of 9 months. These patients should be warned that they are at risk for postoperative complications, and that their ankle plantar flexion strength can remain reduced. PMID- 15782048 TI - Acute traumatic primary patellar dislocation: long-term results comparing conservative and surgical treatment. AB - OBJECTIVE: This report evaluates the subjective, clinical, and functional long term results comparing surgical and conservative treatment in patients with a primary traumatic patellar dislocation. DESIGN AND SETTING: This retrospective clinical study focuses on patients with primary acute traumatic patellar dislocation. Patients with radiologic signs indicative of a predisposition for recurrent patellar instability were excluded from this study. PATIENTS: A total of 126 patients were examined a mean of 8.1 years after initial treatment of their primary patellar dislocation. INTERVENTIONS AND MAIN OUTCOME MEASUREMENTS: Patients were retrospectively divided into groups with conservative therapy (n = 63), diagnostic arthroscopy only (n = 20), immediate surgical reconstruction of the parapatellar ligament complex (n = 37), and refixation of osteochondral fragments (n = 6). Redislocation and resurgery rate, activity level, and subjective, clinical, and functional results were evaluated in these patients, and the outcomes in these groups were compared. RESULTS: In the long term, functional results (as expressed in the Lysholm score) were excellent or good in 85% of the patients, and good subjective results were reported by 71%, but follow up revealed a recurrence rate of 26% in the total study population. The high activity level before the initial trauma could not be completely regained after treatment. There was no significant difference between the surgically and conservatively treated groups in the redislocation and reoperation rates, level of activity, or functional and subjective outcomes. CONCLUSIONS: Even with a focus on acute traumatic etiology and when factors predisposing to recurrent instability are largely excluded, the redislocation rate after treatment of acute patellar dislocation is still high, despite good clinical and subjective results. Conservative management seems to be the treatment of choice in patients with acute patellar dislocation, provided that the generally accepted indications for surgery, such as evidence of osteochondral fragments and major defects of the parapatellar ligament complex, are given due consideration. PMID- 15782049 TI - Effect of ice surface size on collision rates and head impacts at the World Junior Hockey Championships, 2002 to 2004. AB - OBJECTIVE: To determine if collision rates and head impacts in elite junior hockey differed between games played on the small North American ice surface (85 ft wide), an intermediate-size Finnish ice surface (94 ft wide), and the large standard international ice surface (100 ft wide). DESIGN: Videotape analysis of all games involving Team Canada from the 2002 (large ice, Czech Republic), 2003 (small ice, Canada), and 2004 (intermediate ice, Finland) World Junior Championships. All collisions were counted and separated into various categories (volitional player/player bodychecks, into boards or open ice, plus accidental/incidental player/boards, player/ice, head/stick, head/puck). Further subdivisions included collisions involving the head directly or indirectly and notably severe head impacts. RESULTS: Small, intermediate, and large ice surface mean collisions/game, respectively, were 295, 258, 222, total collisions; 251, 220, 181, volitional bodychecks; 126, 115, 88, into boards; 125, 106, 93, open ice; 71, 52, 44, total head; 44, 36, 30, indirect head; 26, 16, 13, direct head; and 1.3, 0.5, 0.3, severe head (P < 0.05 for small-intermediate ice and intermediate-large ice differences in total collisions; P < 0.005 for small-large ice difference; P < 0.05 for small-intermediate ice differences in head impacts; P < 0.01 for small-large ice differences in total and severe head impacts). CONCLUSIONS: There is a significant inverse correlation between ice size and collision rates in elite hockey, including direct, indirect, and severe head impacts. These findings suggest that uniform usage of the larger international rinks could reduce the risk of injury, and specifically, concussions in elite hockey by decreasing the occurrence of collisions and head impacts. PMID- 15782050 TI - A comparison of a preparticipation evaluation history form and a symptom-based concussion survey in the identification of previous head injury in collegiate athletes. AB - OBJECTIVE: The purpose of this study was to compare the incidence of prior head injury reported on preparticipation physical evaluation (PPE) history forms with a retrospective symptom-based survey. DESIGN: A comparison of 2 retrospective survey instruments. SETTING: NCAA Division I varsity athletic program. PARTICIPANTS: A total of 93 male and 79 female athletes participating in intercollegiate contact/collision sports. MAIN OUTCOME MEASURES: Athletes were administered a concussion symptom survey (CSS) with questions about symptom incidence after head injury. These responses were compared with answers given about previous concussion/head injury on the university's PPE history form. The numbers of positive responses were analyzed using descriptive statistics, and differences between the PPE medical history form and the concussion symptom survey were assessed using chi analysis. Factor analysis was performed to assess for possible variance structure between reported symptoms. RESULTS: Seventy-one percent of athletes reporting symptoms consistent with concussion were not identified as having a history of head injury on the PPE medical history form. The most common symptom on the CSS was headache, which accounted for 46 (56.1%) positive responses. CONCLUSIONS: The CSS revealed greater numbers of athletes experiencing symptomatic head injuries than the screening questions on the PPE history form. Screening for signs and symptoms of concussion may enhance the sensitivity of the PPE in detecting a prior history of concussion. PMID- 15782051 TI - Single photon emission computerized tomography and reverse gantry computerized tomography findings in patients with back pain investigated for spondylolysis. AB - BACKGROUND: Athletes with low back pain may have sustained a bone stress injury in the lumbar neural arch, including spondylolysis. Single photon emission computerized tomography (SPECT) and reverse gantry computerized tomography (rg CT) help localize bone stress response and spondylolyses. OBJECTIVES: To describe the SPECT and rg-CT findings in those patients suspected of suffering from a spondylolysis. STUDY DESIGN: Large retrospective case series. SETTING: Secondary and tertiary care center in England. PATIENTS: A total of 213 patients presenting to the Spinal Unit and Sports Medicine Clinics between 1995 and 2000 with low back pain and subsequently investigated for spondylolysis by SPECT and rg-CT. METHODS: An experienced consultant musculoskeletal radiologist reported the SPECT and rg-CT, and these results along with data from the patient's medical records were analyzed. MAIN OUTCOME MEASURES: Sites and sides of increased scintigraphic activity in the lumbar neural arch seen on SPECT and complete and incomplete spondylolyses were recorded. RESULTS: Single photon emission computerized tomography scan showed increased scintigraphic uptake in 145 patients, mostly (42.3%) at L5. Marked increase in scintigraphic uptake was noted on the left side of the neural arch in 31 patients and on the right in 17 (chi; P < 0.05). Spondylolysis was identified on rg-CT in 81 patients. Spondylolyses were bilateral on 44 occasions, but unilateral lesions occurred in the left pars 36 times and on the right 16 times (chi; P < 0.01) CONCLUSIONS: Spondylolyses were a frequent finding on rg-CT in young athletes with low back pain. The sports most commonly associated with this stress response in the pars interarticularis were cricket and soccer. The lower lumbar levels are more likely to show abnormalities on SPECT and computerized tomography. Among those investigated for spondylolysis, left-sided lower lumbar pain was more common than right, and a marked increase in scintigraphic uptake was noted on the left more often than the right. Although bilateral spondylolyses were common, left-sided spondylolyses were more common than right. The asymmetric repetitive movements of certain sports may be responsible. In particular, fast bowlers developed spondylolysis on the opposite side of the body to their bowling arm. PMID- 15782052 TI - Sudden cardiac death in division I collegiate athletics: analysis of automated external defibrillator utilization in National Collegiate Athletic Association division I athletic programs. AB - OBJECTIVE: To determine current outcomes of automated external defibrillator (AED) interventions in sports medicine programs in National Collegiate Athletic Association (NCAA) division I athletics. DESIGN: Qualitative scripted telephone interview with all positive responders to prior NCAA division I-wide study on AED utilization and implementation. SETTING: NCAA division I sports medicine programs. Head athletic trainers were the main data source. PARTICIPANTS: All positive responders to a previously published study on AED implementation in the NCAA division I sports medicine community. Positive responders were those that indicated that they had used their departmental AEDs in a sudden cardiac death (SCD) scenario. MAIN OUTCOME MEASUREMENTS: Survival to hospital discharge was the main outcome sought. When available, additional outcomes were time to defibrillation, time to notification of athletic training staff, EMS response time, location of event, and sudden cardiac victim type (i.e., student, coach, fan). RESULTS: Sixteen departments that previously reported having had an SCD event at their institution responded to this follow-up telephone survey. Twenty percent of AED uses were attributed to student athletes, with 33% of utilizations for athletic department staff and 47% for fans. Defibrillation was actually administered in 53% of AED unit applications. Time to shock was an average of 3.4 minutes, with average EMS response time of 8.2 minutes for those events without EMS on site. Reported survival to hospital discharge in this university athletic department setting for SCD was 0% for students, 75% for staff, 57% for fans, and 61% overall. CONCLUSIONS: The results of this study demonstrate the need for NCAA division 1 athletic sports medicine programs to examine, and possibly expand, the traditional scope of practice of caring primarily for student athletes to include the larger community of sports participants comprised of athletes, departmental staff, and spectators. Athletic department AED programs were extremely successfully at increasing survival of SCD far above national prehospital standards, mainly in the nonathletic population. Further study is also necessary in the realm of AED placement, maintenance, and training of staff. PMID- 15782053 TI - Treatment of stress fractures in athletes with intravenous pamidronate. AB - OBJECTIVE: To determine if the use of a second-generation biphosphonate, pamidronate, is an effective treatment option for stress fractures in intercollegiate athletes. Pamidronate is successfully used to increase bone mass in patients with Charcot arthropathy and osteoporosis. DESIGN: Case series. SETTING: Academic sports medicine clinic with primary responsibility for intercollegiate athletics. PARTICIPANTS: Five intercollegiate female athletes. INTERVENTIONS: Intravenous pamidronate administered weekly for a total of 5 treatments. MAIN OUTCOME MEASUREMENT: Athletes' ability to continue training without restrictions. RESULTS: Four of 5 treated athletes were able to continue training and competition within 1 week of treatment. CONCLUSIONS: Pamidronate may be a useful adjunct to the treatment of stress fractures. PMID- 15782054 TI - Does competitive apnea diving have a long-term risk? Cardiopulmonary findings in breath-hold divers. AB - STUDY PURPOSE: The aim of this study was to investigate the cardiopulmonary status in a competitive apnea diving team. DESIGN: This study was conducted with a cross-sectional study design in which subjects had to undergo a predefined series of cardiopulmonary examinations. SUBJECTS: Eight competitive apnea divers (mean age, 26.9 +/- 5.3 years) who were participating in international apnea diving contests. METHODS: Electrocardiographic, 2-dimensional echocardiographic, Doppler-echocardiographic, and oxymetric examinations were performed in each subject during an apnea test. RESULTS: Apnea diving experience was 2.5 +/- 0.58 years, with a training frequency of 8.9 +/- 6.0 dives per week. During an apnea test, mean apnea time was 4.5 +/- 0.96 minutes. While 2-dimensional echocardiography showed normal morphologic findings of cardiac dimensions and function, electrocardiography, and Doppler echocardiography revealed indicators suggesting the beginning of right ventricular strain. CONCLUSIONS: We interpret the findings of our study as the onset of pulmonary hypertension resulting from repetitive pulmonary vasoconstriction together with severe hypoxia during apnea diving. We conclude that regular competitive apnea diving over a period of >2 to 3 years might carry a chronic cardiopulmonary risk that may lead from early functional changes to manifestation of pulmonary hypertension. PMID- 15782055 TI - Infrapatellar plica as a cause of anterior knee pain. AB - OBJECTIVE: This study seeks to show that in the case of a patient not responding to nonoperative measures for the treatment of anterior knee pain, arthroscopic release of a symptomatic infrapatellar plica can successfully resolve the disability. DESIGN: We report a retrospective study of 12 cases of anterior knee pain not responding to nonoperative treatment that underwent isolated infrapatellar plica resection without other noted knee pathology. SETTING: Patients were evaluated and treated in an outpatient orthopedic sports medicine clinic. PARTICIPANTS: Any patient who presented with anterior knee pain, underwent subsequent arthroscopy, and was treated by isolated resection of the infrapatellar plica was included in the study. INTERVENTION: The surgical procedure involved arthroscopic division of the infrapatellar plica at its attachment on the superior intercondylar notch of the femur. MAIN OUTCOME MEASUREMENTS: Patients were reviewed at least 12 months following the date of surgery. Two subjective knee scales were used to assess knee function. RESULTS: A subjective scale used in prior studies assessing symptomatic medial plica demonstrated 91% percent (11 of 12) excellent (6) or good (5) outcomes at follow up greater than 1 year. The Knee Injury and Osteoarthritis Outcome Scores of knee function on the subscales of pain, symptoms, activities of daily living, sports activity, and quality of life were 97, 96, 99, 99, and 87, respectively. CONCLUSIONS: These cases demonstrate a potential role for the infrapatellar plica as a cause of anterior knee pain. A prospective study is warranted to measure causality. PMID- 15782056 TI - Atraumatic posterior dislocation of the acromioclavicular joint with voluntary reduction. PMID- 15782057 TI - Athlete with traumatic tricuspid regurgitation: return to competition after tricuspid valve repair. PMID- 15782058 TI - Children and running: at what distance safe? PMID- 15782060 TI - Children and marathoning. PMID- 15782061 TI - Preparticipation examination: the Israeli perspective. PMID- 15782062 TI - Exercise duration and intensity in a weight-loss program. AB - OBJECTIVE: To examine the effect of duration and frequency of exercise on weight loss and cardiorespiratory fitness in previously sedentary, overweight, women. DESIGN: Randomized, controlled, 4-arm trial of 12-months duration. SETTING: A university-based behavioral weight loss program during the years 2000 and 2001. PARTICIPANTS: Eligibility criteria were: women, 21 to 45 years of age, body mass index (BMI) 27 to 40, reporting exercise <3 days/week for <20 minutes/day during the previous 6 months. Exclusion criteria were: a history of myocardial infarction, taking medication that would alter the heart rate response during exercise or that would affect metabolism or weight loss, being treated for psychologic conditions, pregnant, recently pregnant, or planning pregnancy, having a medical condition that could affect metabolism or body weight (eg, diabetes) or that would limit exercise participation. INTERVENTION: All 201 participants were assigned to a standard behavioral weight loss program, which included regular group meetings and telephone calls, and caloric and dietary fat restrictions. Participants were given meal plans and kept weekly food diaries. The women were assigned to 1 of 4 exercise groups based on energy expenditure of 1000 kcal/wk or 2000 kcal/wk and exercise intensity (moderate versus vigorous). Exercise intensity was prescribed according to percentage of age-predicted maximal heart rate and rating of perceived exertion. Energy expenditure was converted to minutes of exercise per week. The groups were vigorous intensity/high duration, moderate intensity/high duration, moderate intensity/moderate duration, and vigorous intensity/moderate duration. All 4 groups started the program at moderate intensity and moderate duration (100 min/week of walking) and increased the vigor and duration of exercise to set targets of 200, 300, 200, and 150 min/week, for the groups respectively. Treadmills were provided to the participants, and feedback on their weekly exercise logs was given. MAIN OUTCOME MEASURES: At 6 and 12 months, changes in body weight and BMI were measured. Cardiorespiratory fitness was measured by a graded exercise treadmill test and expressed as percent change in oxygen consumption from baseline. Excluding 5 women who left the study (because of pregnancy or death) the data were analyzed by the intention-to-treat method (completion rate 184/196 = 94%). MAIN RESULTS: Attendance at group sessions and reported dietary intake and exercise adherence suggested that all groups complied similarly with their dietary and exercise prescriptions. Mean weight loss after 1 year was 8.9, 8.2, 6.3, and 7.0 kg, for the vigorous intensity/high duration, moderate intensity/high duration, moderate intensity/moderate duration, and vigorous intensity/moderate duration groups, respectively, but there was no effect of exercise duration or exercise intensity on changes in body weight or in BMI. Cardiorespiratory fitness increased for all groups; 22%, 14.9%, 13.5%, and 18.9%, for the vigorous intensity/high duration, moderate intensity/high duration, moderate intensity/moderate duration, and vigorous intensity/moderate duration groups, respectively, but the groups did not differ in effect of exercise intensity (P = 0.11) or exercise duration (P = 0.35). When participants were divided by their reported average weekly duration of exercise at months 6 and 12, the group which averaged > or =200 min/week at both time points lost more weight than the groups which averaged <150 min/week of physical activity or whose activity duration was inconsistent (difference among groups, P = 0.01). They also had a greater percent increase in cardiorespiratory fitness than those who averaged <150 min/week of physical activity (P = 0.007) and those whose activity was inconsistent (P = 0.003). CONCLUSION: Sedentary overweight women lost weight and improved cardiorespiratory fitness in a year-long combined dietary and exercise regimen. Duration of exercise (at least 150 min/week of walking) was more important than vigorous versus moderate intensity in achieving these goals. PMID- 15782063 TI - Effect of stretching on sport injury risk: a review. AB - OBJECTIVE: Effect of Stretching on Sport Injury Risk: a Review To assess the evidence for the effectiveness of stretching for the prevention of injuries in sports. DATA SOURCES: MEDLINE (1966 to September, 2002), Current Contents, Biomedical Collection, Dissertation Abstracts, the Cochrane Library, and SPORTDiscus were searched for articles in all languages using terms including stretching, flexibility, injury, epidemiology, and injury prevention. Reference lists were searched and experts contacted for further relevant studies. STUDY SELECTION: Criteria for inclusion were randomized trials or cohort studies of interventions that included stretching compared with other interventions, with participants who were engaged in sporting or fitness activities. One author identified 361 articles reporting on flexibility, methods and effects of stretching, risk factors for injury, and injury prevention, of which 6 articles fulfilled the inclusion criteria for meta-analysis. DATA EXTRACTION: Three independent reviewers blinded to the authors and institutions of the investigations assessed the methodologic quality of the studies (100-point scale) and reached consensus on disagreements. Details of study participants, interventions, and outcomes were extracted. Weighted pooled odds ratios were calculated for effects of interventions on an intention-to-treat basis. MAIN RESULTS: Reduction in total injuries (shin splints, tibial stress reaction, sprains/strains, and lower-extremity and -limb injuries) with either stretching of specific leg-muscle groups or multiple muscle groups was not found in 5 controlled studies (odds ratio [OR] 0.93; 95% CI, 0.78 to 1.11). Reduction in injuries was not significantly greater for stretching of specific muscles (OR, 0.80; CI, 0.54-1.14) or multiple muscle groups (OR, 0.96; CI, 0.71-1.28). Combining the 3 ratings of methodologic quality, median scores were 29 to 60/100. After adjustment for confounders, low quality studies did not show a greater reduction in injuries with stretching (OR, 0.88; CI, 0.67-1.15) compared with high quality studies (OR, 0.97; CI, 0.77-1.22). Stretching to improve flexibility, adverse effects of stretching, and effects of warm up were not assessed by appropriate intervention studies. CONCLUSION: Limited evidence showed stretching had no effect in reducing injuries. PMID- 15782064 TI - Past recreational physical activity and risk of breast cancer. AB - OBJECTIVE: To examine the association between baseline and earlier recreational physical activity and incidence of breast cancer in postmenopausal women. DESIGN: Multicenter cohort study. SETTING: Women were enrolled in the Women's Health Initiative Observational Study (WHIOS) at 40 clinical centers between October 1993 and December 1998. PARTICIPANTS: Of a total of 93676 women enrolled in the WHIOS, 74171 were included in this analysis. Eligibility criteria were: aged 50 to 79 years, postmenopausal, and free of serious health conditions that might reduce survival during the following 3 years. Women were excluded if they reported a history of breast cancer or had missing physical activity data. Women of non-European extraction made up 15% of the sample. By February 28, 2002, 3.2% were lost to follow-up and 2.7% had died. ASSESSMENT OF RISK FACTORS: Participants completed baseline questionnaires that included information on medical and reproductive history, and health behaviors including physical activity and diet. Staff collected anthropometric data and information on use of hormone therapy. Women were asked if they usually exercised enough to work up a sweat > or =3 times per week at ages 18, 35, and 50 years. Current (baseline) walking for > or =10 minutes and participation in leisure-time activities were categorized by frequency, duration, and intensity. A current total activity variable was computed from the product of metabolic equivalent values and duration (MET h/wk). MAIN OUTCOME MEASURES: The main outcome measure was the association of incident breast cancer with measures of physical activity during a mean period of follow-up of 4.7 years. Cases of breast cancer were ascertained by the methods of the National Cancer Institute Surveillance, Epidemiology, and End Results guidelines, and collated by physicians and cancer coders blinded to exposure data. MAIN RESULTS: During follow-up, 1780 incident cases of breast cancer were documented. Women who had engaged in strenuous physical activity > or =3 times per week at age 35 had a decreased risk of breast cancer (multivariate adjusted relative risk [RR], 0.86; 95% CI, 0.78-0.95) compared with women who had not. The association was not significant for strenuous activity at age 50 or age 18 years. A greater amount of total current (baseline) physical activity was associated with a lower risk of breast cancer (P for trend, 0.03). Compared with no current physical activity, risk was reduced 18%, 11%, 17%, and 22% for women who exercised 5.1-10, 10.1-20, 20.1-40, and >40 MET h/wk. Hours of current moderate or strenuous physical activity was not significantly related to risk of breast cancer (P for trend, 0.12), although the highest duration category, >7 h/wk, compared with 0 h/wk showed a significant reduction in risk (RR, 0.79; CI, 0.63-0.99). When the women were divided by tertiles of body mass index (BMI cut points at 24.13 and 28.44 kg/m), increased total current activity was protective against breast cancer for those in the lowest tertile of BMI (P for trend, 0.03) but not for those in the upper tertiles (P for trend, 0.74 and 0.30). CONCLUSIONS: More physical activity was associated with a lower risk of breast cancer in postmenopausal American women. An hour every day of moderate or strenuous activity provided most benefit. PMID- 15782065 TI - Topical glyceryl trinitrate for chronic Achilles tendinopathy. AB - OBJECTIVE: To determine whether continuous application of topical glyceryl trinitrate decreases pain and symptoms in chronic noninsertional Achilles tendinopathy. DESIGN: Randomized double-blind placebo-controlled study of 6 months' duration. SETTING: Community and referral study at an Australian University Hospital. PARTICIPANTS: Recruitment was through newspaper advertisements and private consulting rooms. Eligibility criteria were age >18 years, a history of insidious onset of Achilles tendon pain, a tender nodule localized to the region of the calcaneal insertion, and an ultrasound examination that excluded a tendon tear. Exclusion criteria were Achilles tendinopathy of <3 months' duration, a previous operation on, or dislocation of, the affected ankle or leg, distal neurologic signs, a local corticosteroid injection in the previous 3 months, and current pregnancy. The 65 participants (84 affected tendons; 62% men) had a median age of 49 years (range, 24-79 years), with a median duration of symptoms of 16 months (range, 4-147 months). INTERVENTION: Participants were assigned an active transdermal patch (1/4 of a Nitro-Dur 5 [Schering-Plough] glyceryl trinitrate patch), which delivered 1.25 mg of glyceryl trinitrate over 24 hours, or a placebo patch. Patients were required to cut the patches into quarters and apply 1/d to the site of maximal tenderness for the 24-week duration of the study. All patients were also given 500-mg paracetamol tablets for use with headaches, and instructed in a rehabilitation program that comprised rest from aggravating activities, the use of heel-raise wedges, prolonged daily stretching of the gastrocnemius and soleus musculature, and an eccentric calf muscle-strengthening program. MAIN OUTCOME MEASURES: At the baseline, 2, 6, 12, and 24-week examinations the patient completed a symptom assessment sheet to rate the severity of Achilles pain with activity, at rest, and at night (0 = no pain, 4 = very severe pain). The single assessor used the same scale to measure local tenderness; an 11-point scale for the patient to report pain after the single-leg 10-hop test; and also measured the ankle plantar flexor mean peak force and ankle plantar flexor work. Follow-up was 89% complete. MAIN RESULTS: The groups did not differ in pain with activity, night pain, or local tenderness until the 12-week assessment when participants in the glyceryl trinitrate group reported less pain on each measure (mean scores, 0.9 vs. 1.6 [P = 0.02]; 0.2 vs. 0.7 [P = 0.04]; and 0.9 vs. 1.6 [P = 0.02], respectively). The difference was maintained at 24 weeks for pain with activity (mean scores, 0.4 vs. 1.0 [P = 0.03]). At 24 weeks the glyceryl trinitrate group reported less pain on the 10-hop test than the placebo group (mean scores, 0.5 vs. 1.6 [P = 0.005]). Although the intervention group showed a greater increase in plantar flexor mean total work at 24 weeks than the placebo group, the baseline scores were significantly different. The groups did not differ in pain at rest or in ankle plantar flexor peak force. Combining all the measures showed an estimated 14% (95% CI, 9%-19%) excess of asymptomatic tendons in the intervention group at 6 months. Reported main side effects were headaches (glyceryl trinitrate group, 53%; placebo group, 45%) and rashes (glyceryl trinitrate group, 16%%; placebo group, 12%). CONCLUSION: A topical glyceryl trinitrate patch was more effective than placebo for reducing pain from chronic noninsertional Achilles tendonitis in the first 12 and 24 weeks of use. PMID- 15782066 TI - CD56 reactivity in small cell carcinoma of the uterine cervix. AB - Small cell carcinoma (SCC) of the uterine cervix, like its pulmonary counterpart, is a rare but distinctive neoplasm that should be separated from nonendocrine carcinomas because of its highly aggressive clinical course and response to chemotherapy and irradiation. CD56 (neural cell adhesion molecule) has recently been shown to be the best marker for the diagnosis of pulmonary SCC. In this study, we assessed the sensitivity and specificity of CD56 in the diagnosis of SCC of the uterine cervix compared with those of chromogranin and synaptophysin. Twenty-two (88%) of 25 SCCs of the uterine cervix labeled with CD56 in a predominantly membranous and diffuse pattern, whereas 16 of 25 (64%) stained with synaptophysin in a predominantly diffuse pattern and 8 of 25 (32%) showed predominantly focal immunoreactivity for chromogranin. In contrast, 3 of 21 (14%) moderately to poorly differentiated squamous cell carcinomas and 1 of 16 (6%) moderately differentiated adenocarcinomas showed focal immunoreactivity for CD56. Although not specific, CD56 seems to be the most sensitive marker for the diagnosis of SCC of the uterine cervix. Moreover, its diffuse reactivity reduces the possibility of obtaining negative results in small biopsy samples. PMID- 15782067 TI - Immunocytochemical expression of p16INK4A and Ki-67 in cytologically negative and equivocal pap smears positive for oncogenic human papillomavirus. AB - This study was designed to analyze the cross-sectional comparison of the p16 and Ki-67 immunocytochemical expression in negative and equivocal (atypical squamous cells of undetermined significance (ASC-US)) liquid-based cytology (LBC) samples testing positive for high-risk human papillomavirus (HPV) types with HC2 assay or polymerase-chain reaction (PCR). A series of 199 consecutive LBC specimens derived from the same number of women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, Sao Paulo, were analyzed using immunocytochemistry for expression of p16 and Ki-67 in negative and equivocal LBC samples testing positive for high-risk HPV types with hybrid capture II test (HC2) or PCR. All patients with at least one test positive (cytology, PCR, and/or HC2) were followed each 6 months for 3 years. The follow up procedure consisted of visual examination, colposcopic inspection, cytology, and HC2 assay. Among the negative cytologic samples, 101 were HPV-positive and 55 HPV-negative. Of the HPV-positive group, 59 of 101 cases (58.4%) were positive for both p16 and Ki67 immunostaining, and 17 of 101 (16.8%) were negative for both. The proportion of Ki-67-positivity increased almost in parallel with the increasing grade of p16-positivity (p = 0.0001 for linear trend). In the HPV negative group, both markers were negative in 41 of 55 cases (74.5%), and no statistical relationship was observed between the two markers (Pearson, p = 0.595). HPV-positive ASC-US samples demonstrated a simultaneous positive immunoreaction for p16 and Ki67 in 11 of 16 cases (68.7%), whereas 3 (18.7%) were concurrently negative. The relationship between the two markers was of borderline significance (Pearson, p = 0.053), but no linear relationship was found between the graded p16 and Ki-67 expression (p = 0.065 for linear trend). In the HPV negative ASC-US group, there was no statistical association between the graded p16 and Ki-67 positivity (Pearson, p = 0.281). After 36 months of follow-up of the ASC-US patients, 6 women still displayed ASC-US smear, of which 4 of 6 were HPV-positive and expressed both p16 and Ki-67 markers. Two of 43 ASC-US smears had high-grade squamous intraepithelial lesions diagnosed (4.6%), and 1 had low grade squamous intraepithelial lesion (2.3%). All of those were positive for HPV, p16 and Ki-67. Patients with ASC-US diagnosis and positive high-risk HPV status and positive for p16 Ki67 should be carefully observed to exclude occurrence of a squamous intraepithelial lesion. The combination of these two markers can be a useful implement for management of women with equivocal cytology. PMID- 15782068 TI - The relationship of glands to thick-wall blood vessels as a marker of invasion in endocervical adenocarcinoma. AB - The recognition of early invasion in endocervical adenocarcinomas can be difficult. We evaluated the proximity of endocervical glands and their pattern of infiltration in relation to thick-wall blood vessels as potential markers of invasion in a series of invasive endocervical adenocarcinomas removed by cone biopsy or hysterectomy. Routinely stained slides were examined from 50 invasive endocervical adenocarcinomas (37 of usual type and 13 of minimal deviation type), 26 noninvasive lesions (14 cases of adenocarcinoma in situ, 7 cases of hyperplasia, 4 cases of tunnel clusters, 1 adenomyoma), and 20 normal cervices, including 7 with deep nabothian cysts. The range of vessel wall thickness using an ocular micrometer was recorded in each specimen. A blood vessel with a wall > or =36-microm thick was defined as thick-walled. Both the blood vessel wall thickness and distance to the closest gland was recorded for each case. The median distance from a thick-wall blood vessel to a gland in the invasive tumors was 30 microm compared with 168 microm in the noninvasive group. In both groups, the measured blood vessel wall had a median thickness of 42 microm. The pattern of infiltration of glands around the thick-wall vessels was classified as "circumferential," in which multiple glands circumferentially enveloped the vessel, or "molded," in which single or multiple distorted glands were molded around the vessel. The circumferential and molded patterns were identified in 35 (70%) and 10 (20%) of 50 invasive adenocarcinomas, respectively. Of the 26 lesions in the noninvasive group, only 3 (6%) had a circumferential pattern and none had a molded pattern. Importantly, none of the cases in the noninvasive group showed either pattern in the presence of glandular atypia. In conclusion, close proximity of glands to thick-wall blood vessels (distance from the closest gland to a thick-wall vessel less than or equal to the thickness of the vessel wall) seems to be a useful feature in the diagnosis of invasive endocervical adenocarcinoma. This feature, in combination with certain glandular growth patterns and cytologic atypia, can be highly suggestive, if not diagnostic, of invasive carcinoma. PMID- 15782069 TI - TFE3 immunoreactivity in alveolar soft part sarcoma of the uterine cervix: case report. AB - SUMMARY: : Alveolar soft part sarcoma (ASPS) is an uncommon neoplasm that has only rarely been reported in the female genital tract. We report a case of incidentally discovered ASPS in the uterine cervix of a 39-year-old woman that exhibited immunoreactivity for TFE3, a recently described marker of ASPS. Strong nuclear immunoreactivity for TFE3 in tumors characterized by a chromosomal translocation involving the TFE3 gene has been reported in ASPS and a subset of pediatric renal cell carcinomas. This translocation, t(X;17)(p11;q25), fuses the TFE3 transcription factor gene on Xp11 to a novel gene on 17q25, designated ASPL. In our case, the neoplastic nuclei were strongly positive for TFE3. To the best of our knowledge, this is the first report of ASPS of the female genital tract with immunoreactivity for TFE3. Other interesting findings in our case were the small size of the tumor and the absence of clinical symptoms. PMID- 15782070 TI - Mucinous ovarian tumors of Mullerian-type: an analysis of 17 cases including borderline tumors and intraepithelial, microinvasive, and invasive carcinomas. AB - Mullerian-type mucinous tumors (MMTs) of the ovary are characterized by a papillary architecture similar to that of serous tumors and a content of endocervical-like mucinous epithelium. The latter may be admixed with other mullerian-type epithelia, including those of serous, endometrioid, and squamous types, and indifferent eosinophilic epithelial cells. We analyzed 17 MMTs, including 12 borderline tumors, 2 intraepithelial carcinomas, 2 microinvasive carcinomas, and 1 invasive carcinoma. Fourteen of 16 tumors (88%) with available staging were stage I; the remaining two cases (both borderline) were stage IIa and IIIc. Endometriosis was identified in eight cases (47%). Only two patients (12%) had bilateral tumors, a frequency of bilaterality lower than in previous studies. Five patients (31%) had conservative treatment consisting of a cystectomy or unilateral salpingo-oophorectomy. All patients had a favorable outcome, with no recurrences or disease-related deaths, regardless of the presence of high mitotic index, intraepithelial carcinoma, microinvasion, bilaterality, conservative treatment, or advanced stage. This indolent behavior of MMTs is similar to that previously reported, but additional cases of invasive carcinomas in this category are needed to better define their outcome. PMID- 15782071 TI - Assessment of Her-1, Her-2, And Her-3 expression and Her-2 amplification in advanced stage ovarian carcinoma. AB - The human epidermal growth factor receptor (Her) family of receptor tyrosine kinases includes Her-1, Her-2, and Her-3. The overexpression of Her-1 and Her-2 have been reported previously in surface epithelial ovarian cancer. Although up to one-third of ovarian carcinomas have been found to have amplification or overexpression of Her-2, responses to trastuzumab therapy in these patients have been disappointing. In this study, we examined Her-1, Her-2, and Her- 3 protein expression as well as the frequency of Her-2 amplification in a series of 103 high-grade, advanced-stage (FIGO stage III or IV) ovarian surface epithelial carcinomas. Immunohistochemical staining using commercially available antibodies against Her-1-3 and fluorescence in situ hybridization (FISH) using probes against Her-2 and chromosome 17 centromere (CEP) were performed on a tissue microarray containing cores of tumor from 103 surface epithelial carcinomas (85 serous, 6 mixed surface epithelial, 5 clear cell, 3 endometrioid, 3 undifferentiated, 1 mucinous). Nine of 99 (9.1%) tumors were positive for Her-1 expression and 5 of 102 (4.9%) tumors were positive for Her-2 expression, with 1 showing strong immunoreactivity. None of the Her-1 positive tumors exhibited Her 2 immunoreactivity. There was no correlation between Her-1 or Her-2 expression and survival. Using Her-2:centromere fluorescence ratios of 2.0 or 1.5 as cutoffs in assessment of Her-2 amplification, 8 of 75 (10.7%) and 25 of 75 (33.3%) tumors, respectively, showed Her-2 amplification. Two of eight tumors that showed higher level (>2) Her-2 amplification by FISH also were positive for Her-2 by immunohistochemistry. Only 3 of 103 tumors expressed Her-3. Immunoreactivity for Her-1 and Her-2 was less frequently observed in this series than has been previously reported. The strong correlation between Her-2 immunostaining and amplification characteristic of breast carcinoma is not seen in ovarian carcinoma. These results indicated that few patients with ovarian carcinoma have tumors that would benefit from therapy targeted specifically against Her-1, Her 2, or Her-3. PMID- 15782072 TI - Immunolocalization of nuclear transcription factors, DAX-1 and Ad4BP/SF-1, in human common epithelial ovarian tumors: correlations with StAR and steroidogenic enzymes in epithelial ovarian carcinoma. AB - Intratumoral steroidogenesis is considered to play important roles in the biologic behavior of common epithelial ovarian carcinoma. Recent studies have demonstrated the important roles of Ad4BP/SF-1 and DAX-1 in regulation of steroidogenesis. In this study, we evaluated DAX-1, Ad4BP/SF-1, StAR, and steroidogenic enzyme expressions and their correlations in epithelial ovarian carcinomas to evaluate the possible roles of these factors in regulation of intratumoral steroid metabolism and/or production. Immunolocalization of DAX-1, Ad4BP/SF-1, StAR, and steroidogenic enzymes were examined in 90 epithelial ovarian carcinomas. mRNA expression of these proteins was evaluated using real time polymerase chain reaction (PCR) in 23 cases for further characterization. DAX-1 and Ad4BP/SF-1 immunoreactivity was detected predominately in the nuclei of tumor cells, whereas that of StAR and steroidogenic enzymes was present in the cytoplasm. We detected a significant positive correlation between StAR and steroidogenic enzymes immunoreactivity and Ad4BP/SF-1 and statistically inversed correlation with DAX-1. A positive statistical correlation was detected between intratumoral stromal Ad4BP/SF-1 immunoreactivity and clinicopathologic parameters of carcinoma patients. Results of real-time PCR analysis were correlated with those of immunohistochemical studies. The status of intratumoral DAX-1, Ad4BP/SF 1, and StAR and steroidogenic enzymes in epithelial cells and intratumoral stromal cells of epithelial ovarian carcinoma may contribute in the progression and/or aggressiveness of these tumors. PMID- 15782073 TI - Initial endometriosis showing direct morphologic evidence of metaplasia in the pathogenesis of ovarian endometriosis. AB - It is believed that ovarian endometriosis may be generated by a celomic metaplastic process from existing epithelium in the ovary. However, no morphologic evidence of metaplastic process has been described. In this study, we intended to identify the earliest morphologic changes of endometriosis within the ovary to examine if evidence of metaplasia exists. Included in this study were 110 ovarian endometriosis cases and 30 benign ovaries without endometriosis but with ovarian epithelial inclusions (OEIs). Among the 110 well-established ovarian endometriosis cases, 34 cases showed areas of initial endometriosis (IE), which is defined as lesions showing direct transitions from normal-looking ovarian tissue to areas of minimal formation of endometriosis and/or to areas of full blown endometriosis. We further divided IE into two types: type I IE was present on the ovarian surface, which was associated with ovarian surface epithelia; type II was located within the ovarian cortex, which was associated with OEIs. Sections containing IE, OEIs, and well-formed endometriosis were subject to CD10 and aromatase immunostaining. In IE lesions, the number of CD10-positive cells were significantly higher than the number of that in OEIs, but lower than that of well-formed endometriosis areas (p < 0.05). Aromatase expression was detected in both epithelial and stromal components of the IE lesions, indicating that estrogen local production may be involved in this initial process of endometriosis. Microvessel density was higher in IE lesions than in areas of OEI (p < 0.05). Based on the morphologic characteristics of IE, we believe that IE represent a spectrum of the earliest morphologic changes of endometriosis identifiable by routine microscopy. The morphologic transitions from ovarian surface epithelium or OEI to IE lesions provide direct metaplastic evidence for the pathogenesis of ovarian endometriosis. This metaplastic process may not only involve the ovarian epithelial cells, but also stromal components. Local production of estrogen, probably in high-levels, may be related to the initial process of endometriosis, although detailed mechanisms remain to be clarified. PMID- 15782074 TI - Diagnostic challenge of fetal ontogeny and its application on the ovarian teratomas. AB - Although neuroepithelial tubules (NET) often are a component of immature teratoma (IT), they are not always required for diagnosis. Other somatic elements are sufficient and often verified with immunohistochemical stain. This study was designed to determine the definition of immaturity versus fetal ontogeny, using several molecular markers in IT. It is our contention that IT is equivalent to an embryonic stage less than a fertilization age (FA) of 8 weeks, and a mature teratoma (MT) to a fetal stage later than a FA of 8 weeks, whereas an embryonal carcinoma (Eca) matches a pre-embryonic stage earlier than a FA of 2 weeks. The teratomatous components used as a roadmap to evaluate maturity included: a lobular structure of primitive endodermal tubules (FA 4 to 6 weeks), a ventricle lined cortical plate (FA 9 weeks), a complex papillary choroid plexus (FA 10 weeks), melanin deposition in hair follicles (FA 15 weeks), and the bell stage of odontogenesis (FA 19 weeks). The teratomatous components of 25 resected ovarian solid teratoma samples were compared with fetal ontogeny. For an immunohistochemical analysis, the CD30, CD34, CD99, bcl-2, alpha-fetoprotein (AFP), and placenta-like alkaline phosphatase (PLAP) were assessed. The AFP and Ki-1 were positive in the embryoid body, which was identified at a FA less than 4 weeks in Eca. The AFP was positive in the primitive endodermal components and some of the squamous epithelium in IT. The CD99 and bcl-2 were selectively stained in the primitive NET, which was detected no later than a FA of 6 weeks. The CD34 and bcl-2 were positive in the immature-looking precartilage blastomatous components, which proved useful for detecting immature cartilage, corresponding to a FA of 5 to 6 weeks. The ontogeny of IT was found to correspond to the embryonic stage at a FA of 2 to 8 weeks, and CD99, CD34, bcl-2, AFP, CD30, and PLAP could be used as supportive tools to define IT. This new grading system could be more scientific and more reproducible in any spectra of teratoma. PMID- 15782075 TI - Immunohistochemical detection of human telomerase reverse transcriptase (hTERT), topoisomerase IIalpha expression, and apoptosis in endometrial adenocarcinoma and atypical hyperplasia. AB - This study evaluated the immunohistochemical expression of human telomerase reverse transcriptase (hTERT) in endometrial carcinoma and atypical endometrial hyperplasia, and related it to the expression of topoisomerase (TP)IIalpha (a proliferation associated enzyme); apoptosis as determined by the frequency of apoptotic bodies (ABI); mitotic counts; and other clinicopathologic variables. Immunoreactivity for hTERT and TPIIalpha as well as ABI were assessed in 57 endometrial samples (12 atypical hyperplasias, 33 endometrioid carcinomas, 12 serous/clear cell carcinomas). hTERT immunoreactivity, TPIIalpha labeling indices (LI), ABI, and ratios of the indices (ABI/TPIIalpha LI) increased from atypical hyperplasias to endometrioid carcinomas to serous/clear cell carcinomas (p < 0.0001 for each variable). hTERT expression increased with ABI (p < 0.0001), TPIIalpha LI (p = 0.0019), ABI/TPIIalpha ratios (p < 0.0001), and grade (p = 0.0005), but not with FIGO stage (p = 0.2775). TPIIalpha LI, ABI, and ratios were related to high grade (p = 0.0001 for each variable), but not with FIGO stage (p = 0.7362, p = 0.7554, and p = 0.7405, respectively). TPIIalpha LI and ABI were significantly correlated in atypical hyperplasias (p = 0.0004), endometrioid carcinomas (p < 0.0001), and serous/clear cell carcinomas (p = 0.024). Immunostaining levels for hTERT were similar in atypical hyperplasias and grade 1 endometrioid carcinomas (p = 0.1956). These results suggest that hTERT expression is closely related to proliferation, apoptosis, and high grade in endometrial carcinomas, reflecting cell cycle deregulation in endometrial carcinogenesis. PMID- 15782076 TI - Ectopic sebaceous glands in the vagina. AB - We report a case of ectopic sebaceous glands in the upper third of the posterior wall of the vagina of a 49-year-old woman. This is, to our knowledge, the first fully documented case report of their occurrence in the vagina. The review of the literature and possible embryologic origin are discussed. PMID- 15782077 TI - Lipomatous variant of angiomyofibroblastoma: report of two cases and review of the literature. AB - Angiomyofibroblastoma (AMF) is a rare, benign, mesenchymal tumor occurring mainly in the female genital tract. Some cases contain scattered mature adipocytes, but the lipomatous variant in which mature adipose tissue is prominent or striking is rare. Only five cases have been reported in the English literature. We report two more such cases that were composed of 70 to 80% and 30 to 40% adipose tissue, respectively. Immunohistochemical analysis showed that the tumor cells were positive for estrogen receptor, progesterone receptor, vimentin, and Bcl-2, and negative for cytokeratin AE1/1, EMA, and CD117. Tumor cells in the first case were positive for CD34 but not desmin and muscle-specific actin. The opposite expression profile of these three markers was observed in tumor cells in the second case: positive for desmin and muscle-specific actin and negative for CD34. Rare cells were positive for S-100 in adipose-rich areas in the first case. Our findings indicate that the tumor cells in the lipomatous variant have similar immunoprofile to those of usual AMF and support the concept that the lipomatous variant probably represents an extreme end of a wide spectrum of differentiation in AMF. PMID- 15782078 TI - Intravascular large B-cell lymphoma of the uterus: a diagnostic challenge. AB - A 63-year-old woman presented with postmenopausal bleeding. The initial curettage material was reported as "undifferentiated malignant tumor." A total abdominal hysterectomy with bilateral salpingo-oophorectomy was performed with pelvic lymph node dissection and omental biopsy, which showed an intravascular large B-cell lymphoma in the lumina of small blood vessels of the uterus, left tube, and right ovary. This is the first reported case of an intravascular lymphoma of the uterus. PMID- 15782079 TI - Ten-year review of rating scales, VII: scales assessing functional impairment. AB - OBJECTIVE: This is the seventh in a series of 10-year reviews of rating scales. Here the authors present scales measuring functional impairment, a sequela of mental illness. The measurement of functional impairment has assumed importance with the recognition that symptom resolution does not necessarily correlate with functional improvement. METHOD: The authors reviewed functional impairment from multiple sources over the past 20 years. Thus, this article includes a variety of scales ranging from those that have been subject to critical review with strong psychometric support to those that have not been critically reviewed but are in widespread use to those that are still finding their niche. RESULTS: These scales represent a continuum of constructs from symptoms to functional impairment to contextual factors that affect youths' functioning. Most older scales have focused on developmentally delayed youths. Newer scales strive to measure functional impairment separate from symptomatology. Some newer scales are also keyed to determination of level of service need. CONCLUSIONS: Scales measuring functional impairment can elucidate the impact of illness on youths, identify targets for treatment, determine service needs, and monitor treatment effectiveness. These scales are widely used in community mental health and health service delivery. They can assist in providing evidence-based treatment. PMID- 15782080 TI - Commentary: using scales assessing functional impairment in supplemental security income eligibility determination. PMID- 15782081 TI - An open-label trial of escitalopram in pervasive developmental disorders. AB - OBJECTIVE: To assess the effect of escitalopram in the treatment of pervasive developmental disorders (PDDs). METHOD: This 10-week study had a forced titration, open-label design. Twenty-eight subjects (mean age 125.1 +/- 33.5 months) with a PDD received escitalopram at a dose that increased weekly to a maximum dose of 20 mg as tolerated. The Aberrant Behavior Checklist-Community Version (ABC-CV) and the Clinical Global Impression scale (CGI) were used to assess outcome. RESULTS: There was significant improvement in ABC-CV Irritability Subscale Scores (baseline mean 20.5 +/- 5.9 to final mean 10.9 +/- 7.2; p < or = .001) and in the other ABC-CV Subscales. Improvement on Clinical Global Improvement Scale severity rating was also significant (baseline mean 5.2 +/- 1.0 to final mean 4.6 +/- 1.2; p < or = .001). Twenty-five percent of the subjects responded at a dose less than 10 mg and did not tolerate the 10-mg dose, and an additional 36% responded at a dose greater than or equal to 10 mg. Final dose was unrelated to weight and only weakly correlated with age. CONCLUSIONS: This open label study found escitalopram to be useful in treating some difficulties common in PDDs. A wide variability in dose was found that could not be accounted for by weight and only partially by age. The study provides information useful for the design of double-blind, placebo-controlled studies of escitalopram in PDDs. PMID- 15782082 TI - Steady-state pharmacokinetics of bupropion SR in juvenile patients. AB - OBJECTIVE: To examine the steady-state pharmacokinetic properties of bupropion sustained release (SR) and their potential developmental differences in youths. METHOD: Eleven boys and eight girls aged 11 to 17 years old were prescribed bupropion SR monotherapy for attention-deficit/hyperactivity disorder (n = 16) and/or depressive disorders (n = 16). Bupropion SR was given in morning doses of 100 mg/day (n = 11) or 200 mg/day (n = 8) for 14 days or less, with five subjects studied on both doses. All subjects had blood draws from an intravenous port every 1 to 3 hours for 24 hours after their usual morning doses. Pharmacokinetic variables were determined by noncompartmental and compartmental analyses for bupropion and metabolites, respectively. RESULTS: Bupropion and its metabolites exhibited linear pharmacokinetics. Areas under the concentration curves for the hydroxybupropion, threohydrobupropion, and erythrohydrobupropion were 20, 12, and 2.7 times higher, respectively, than for bupropion. Relative to adults, the mean half-lives of bupropion (12.1 hours) and threohydrobupropion (26.3 hours) were significantly shorter, and areas under the concentration curve ratios of metabolites to bupropion were 19% to 80% higher. CONCLUSIONS: Youths metabolize bupropion SR faster to hydroxybupropion and other active metabolites than adults. Until the clinical importance of bupropion's metabolites is clarified, bupropion SR should be given in divided doses to youths, as the manufacturer recommends for adults taking higher doses. PMID- 15782083 TI - Early to bed: a study of adaptation among sexually active urban adolescent girls younger than age sixteen. AB - OBJECTIVE: To examine the association between sexual activity among urban adolescent girls and four global measures of psychosocial adaptation (academic motivation, school achievement, depressive symptoms, and expectations about the future). METHOD: Data derived from the Social and Health Assessment, a self report survey administered in 1998 to students in the public school system in New Haven, CT (149 classes at 17 middle and high schools). RESULTS: Of 1,413 respondents (57% black, 28% Hispanic; mean age 13.4 +/- 1.7 years), 414 (29%) acknowledged prior sexual intercourse; the proportions of sexually active girls in 6th, 8th, and 10th grades were 14%, 30%, and 50%, respectively. In multivariate analyses of covariance, sexual activity was significantly associated with all four measures of psychosocial adaptation (p < .001). Other correlates of at least one measure of maladaptation included socioeconomic status, sensation seeking, and lower school grade (p < .001 for each), peer pressure (p < .01), and black ethnicity, and the interaction of sexual activity by lower school grade (p < .05 for each). CONCLUSIONS: Compared with their sexually naive peers, sexually active adolescent girls had lower scores on global measures of psychosocial adaptation. These findings have clinical, policy, and research relevance to a vulnerable population at high risk of teenage pregnancy and sexually transmitted diseases. PMID- 15782084 TI - Gender differences in ADHD subtype comorbidity. AB - OBJECTIVE: To examine gender differences in attention-deficit/hyperactivity disorder ("ADHD") symptom comorbidity with "oppositional defiant disorder", "conduct disorder", "separation anxiety disorder", "generalized anxiety disorder", speech therapy, and remedial reading in children. METHOD: From 1994 to 1995, data from a large sample (N = 4,371) of twins and siblings studied in the Australian Twin ADHD Project were obtained by mailed DSM-IV-based questionnaires, investigating patterns of comorbidity in the three subtypes of "ADHD": "inattentive", "hyperactive/impulsive", and "combined". A total of 1,550 questionnaires were returned (87%) over the next 12 to 18 months. RESULTS: Analysis of variance showed significant between-group differences in males and females for inattention and hyperactive/impulsive symptom counts with higher rates of "oppositional defiant disorder" and "conduct disorder" in males, and higher rates of "separation anxiety disorder" in females indicating internalizing disorders are more common in females and externalizing disorders are occurring more often in males. Differences were found between the "ADHD" subtypes and the no ADHD category for all comorbid conditions, for both males and females. Children without ADHD consistently had fewer symptoms, while children with the combined subtype showed consistently more comorbid symptoms indicating a strong relationship between high rates of externalizing symptoms and high rates of internalizing symptoms. Gender differences in speech therapy were significant only for the children without ADHD. The rates of "separation anxiety disorder" were higher in females with the "inattention" subtype and the rate of "generalized anxiety disorder" higher for females with the "combined" subtype, indicating that the subtypes of ADHD were associated with these internalizing disorders in different ways. CONCLUSIONS: Although comorbidity differs among ADHD subtypes, there were no significant gender differences in comorbidity for externalizing disorders. Inattentive girls may present with anxiety. Clinical approaches for both males and females should be sensitive to possible language and reading problems. PMID- 15782085 TI - A meta-analysis of working memory impairments in children with attention deficit/hyperactivity disorder. AB - OBJECTIVE: To determine the empirical evidence for deficits in working memory (WM) processes in children and adolescents with attention-deficit/hyperactivity disorder (ADHD). METHOD: Exploratory meta-analytic procedures were used to investigate whether children with ADHD exhibit WM impairments. Twenty-six empirical research studies published from 1997 to December, 2003 (subsequent to a previous review) met our inclusion criteria. WM measures were categorized according to both modality (verbal, spatial) and type of processing required (storage versus storage/manipulation). RESULTS: Children with ADHD exhibited deficits in multiple components of WM that were independent of comorbidity with language learning disorders and weaknesses in general intellectual ability. Overall effect sizes for spatial storage (effect size = 0.85, CI = 0.62 - 1.08) and spatial central executive WM (effect size = 1.06, confidence interval = 0.72 1.39) were greater than those obtained for verbal storage (effect size = 0.47, confidence interval = 0.36-0.59) and verbal central executive WM (effect size = 0.43, confidence interval = 0.24-0.62). CONCLUSION: Evidence of WM impairments in children with ADHD supports recent theoretical models implicating WM processes in ADHD. Future research is needed to more clearly delineate the nature, severity, and specificity of the impairments to ADHD. PMID- 15782086 TI - Familial clustering of executive functioning in affected sibling pair families with ADHD. AB - OBJECTIVE: To investigate familial clustering of executive functioning (i.e., response inhibition, fine visuomotor functioning, and attentional control) in attention-deficit/hyperactivity disorder (ADHD)-affected sibling pairs. METHOD: Fifty-two affected sibling pairs aged 6 to 18 years and diagnosed with ADHD according to DSM-IV performed the Stroop test, go/no-go task, two different fine visuomotor tracking tasks, and a sustained-, divided-, and focused attention task. RESULTS: Significant correlations (r = 0.4) were found between siblings for response inhibition and attentional control and for fine visuomotor skills that made high demands on executive functioning. CONCLUSIONS: Response inhibition, higher order controlled fine visuomotor functioning, and attentional control seem to cluster in ADHD-affected siblings. This suggests that these aspects of executive dysfunctioning may reflect an endophenotype of ADHD. Measurement of these executive functions may facilitate the identification of genes involved in ADHD by forming more homogeneous subgroups. PMID- 15782087 TI - Delirium in severely ill young children in the pediatric intensive care unit (PICU). PMID- 15782088 TI - Commentary: the diagnosis of delirium in pediatric patients. PMID- 15782089 TI - A simple method using a polymethylpenten chamber for isolation of human pancreatic islets. AB - OBJECTIVES: Isolation of large numbers of intact and functional human islets remains difficult and expensive. We describe a novel method using a polymethylpenten chamber (PMPC) and compare its efficacy to the classic method using a stainless steel chamber (SSC). METHODS: Five pancreases obtained from cadaveric donors were processed with the SSC method, and the islets were purified with a Cobe cell separator. The next 15 pancreases (similar donor characteristics) were distended with Liberase HI, minced, and digested in a PMPC whose thermic properties did not require continuous heating to maintain temperature of the prewarmed medium at 37 degrees C. The digestion was done in 2 phases to avoid damaging the first freed islets. Digested tissue was filtered on a column of 6-mm glass beads and 500-microm mesh screen, so that tissue volume was small enough to permit purification on discontinuous Ficoll gradients in tubes. RESULTS: With the PMPC method, the extent of digestion (+/-70%), yield (approximately 5000 IEQ/g), and final purity (73%) and viability (84%) of the islets was similar to those with the SSC, but the proportion of large islets (>150 microm in diameter) was higher. Cell composition (beta vs. non-beta cells) of isolated islets was not different from that of islets in situ in the same pancreas. Islet function, assessed by perifusion, showed an excellent average stimulation index of approximately 13-fold (15 vs. 1 mmol/L glucose, without cAMP raising agent). CONCLUSIONS: This new method for isolation of human islets uses simple, low-cost, and potentially disposable material and requires a team of only 2 persons. The technique is as efficient as the classic SSC method and provides islets with excellent integrity and insulin-secreting capacity. PMID- 15782090 TI - Dorsal pancreatic duct dominance in pancreaticobiliary maljunction. AB - OBJECTIVES: In patients with pancreaticobiliary maljunction (PBM), reflux of pancreatic juice to the bile duct may contribute to carcinogenesis of the biliary tract. This study aimed to investigate the pancreatographic findings in patients with PBM and the relationship to their clinical findings in view of pancreatic juice flow. METHODS: Seventy-eight cholangiopancreatograms were reviewed. When the maximum diameter of the Santorini duct was almost equal to or greater than that of the ventral pancreatic duct, the relationship between the 2 ducts was defined as dorsal pancreatic duct (DPD) dominance. RESULTS: Pancreatographic findings were divided into 2 groups: a normal duct group (69 patients) and a DPD dominant group (9 patients). Although 40 patients (58%) with biliary carcinoma were identified in the normal duct group of PBM, only 1 gallbladder carcinoma (11%) occurred in DPD-dominant patients (P < 0.01). A large-caliber Santorini duct was noted to flow straight from the upstream DPD in all patients with DPD dominance. Concentration of amylase in the bile of DPD dominance was significantly lower than that of normal pancreatic duct system (P < 0.01). CONCLUSIONS: In PMB with DPD dominance, most pancreatic juice in the upper DPD is drained into the duodenum through the minor duodenal papilla, and reflux of pancreatic juice to the biliary tract might be reduced, resulting in reduced frequency of associated biliary carcinogenesis. PMID- 15782091 TI - FTY720, an immunosuppressant, attenuates chronic pancreatitis in rats by suppressing T-cell infiltration. AB - OBJECTIVES: FTY720, a novel synthetic immunosuppressant, decreases peripheral blood lymphocytes by accelerating their homing to the peripheral and mesenteric lymph nodes and Peyer's patches. We previously reported that tacrolimus, another immunosuppressant, attenuates chronic pancreatitis by suppressing T-cell infiltration in male Wistar Bonn/Kobori rats but also may cause toxicity. To assess the effects of FTY720 on the development of pancreatic inflammation and fibrosis in the same model, the agent dissolved in physiologic saline was subcutaneously injected to 10-week-old male WBN/Kob rats for 10 weeks. METHODS: Parameters for inflammation and fibrosis were assessed and interferon-gamma and transforming growth factor-beta1 mRNA in the pancreas were determined by RT-PCR. RESULTS: Treatment with FTY720 attenuated gross alterations in the pancreas, including pigmentation and atrophy. This protective effect was quantitatively confirmed by significant increase in pancreatic weights and decreases in pancreatic myeloperoxidase activity (an index of granulocyte infiltration), pancreatic hydroxyproline content (an index of collagen deposition), ratio of fibrous tissue, and histologic scores. The obvious infiltration of CD4- and CD8 positive T cells into the pancreas in the saline group was almost completely prevented by administration of FTY720, which also suppressed overexpression of interferon and transforming growth factor-beta1 mRNA in the pancreas. CONCLUSION: We conclude that FTY720 prevents pancreatic inflammation and fibrosis by suppressing infiltration of CD4- and CD8-positive T cells and by downregulating induction of interferon and transforming growth factor-beta1 mRNA in the pancreas. PMID- 15782092 TI - Amelioration of pancreatic fibrosis in mice with defective TGF-beta signaling. AB - OBJECTIVES: Pancreatic fibrosis is a characteristic feature of chronic pancreatic injury, which is a result of the imbalance between synthesis and degradation of extracellular matrix (ECM) proteins. Transforming growth factor-beta (TGF-beta) plays a central role in biosynthesis and turnover of the ECM. In this study, we evaluated the role of TGF-beta signaling in pancreatic fibrosis induced by repetitive acute pancreatic injuries with mice of dominant-negative mutant of TGF beta receptor II selectively in pancreas. METHODS: TGF-beta signaling was inactivated by overexpressing a dominant-negative mutant form of TGF-beta type II receptor (pS2-dnR II) only in the pancreas under control of pS2/TFF1 promoter. Pancreatic fibrosis was induced by repeated intraperitoneal injections of 40 microg/kg cerulein for 5 or 10 weeks. RESULTS: Repeated administration of cerulein induced significant pancreatic fibrosis, but of which fibrosis was remarkably attenuated in pS2-dnR II mice compared with wild-type littermates (P < 0.01). The ameliorated fibrosis was due to the reduction of synthesis of ECM proteins such as collagen type I, fibronectin, and ICAM-1. DNA binding activity of transcriptional factors including nuclear factor (NF)-kappaB and AP-1, responsible for the induction of immediate early genes of inflammatory responses, were significantly decreased in pS2-dnR II mice. While TGF-beta1 treatment in isolated pancreatic stellate cells (PSCs) stimulated the expression of alpha-SMA and fibronectin, PSCs transfected with TGF-beta dnRII showed attenuation of the ECM components. CONCLUSION: Conditional loss of TGF-beta signaling selectively in the pancreas led to a failure in fibrogenic responses of repeated injections of cerulein, signifying that the modulation of TGF-beta signaling could be the therapeutic target for the prevention of chronic fibrosing pancreatitis. PMID- 15782093 TI - Severe acute pancreatitis: nonsurgical treatment of infected necroses. AB - OBJECTIVES: In acute pancreatitis, infection of necrosis is associated with a substantial mortality of 15% to >50% even if immediate necrosectomy, the recommended standard treatment, is performed, mainly because of the patients' critical systemic and unstable local conditions at the time of manifestation of infection. We investigated whether this dreaded complication can be managed conservatively. METHODS: We evaluated 88 consecutive patients with severe (APACHE II score, > or =11; Ranson score, > or =4) acute necrotizing pancreatitis who received ICU treatment including early antibiotic prophylaxis. Twenty-eight patients were included who developed infection of necroses, verified by fine needle aspiration, 19 +/- 6 days after admission. No patient received urgent surgery; rather, in all patients, nonsurgical therapy was continued after adapting the antibiotic regimen to bacteriology. In the further course, 12 patients were excluded due to refractory local complications eventually requiring surgical treatment 36 +/- 14 days after diagnosis of infection. RESULTS: Sixteen patients (APACHE II score: 18.1 [11-33]; Ranson score, 5.9 [4-10]) were managed with medical treatment alone. Six patients recovered without further complications; 10 patients (62%) developed single or multiple organ failure, and 2 died (mortality, 12%). CONCLUSION: These data suggest that in patients with acute necrotizing pancreatitis and infected necroses, surgery can be avoided without compromising prognosis and outcome. PMID- 15782094 TI - Prognostic predictive value of endoscopic ultrasound findings for invasive ductal carcinomas of pancreatic head. AB - OBJECTIVES: Accurate preoperative prediction of the prognosis of patients with invasive ductal carcinoma of pancreatic head (pancreatic head cancer) is important for selecting treatment methods. We retrospectively examined the prognostic predictive values of endoscopic ultrasound (EUS) findings for patients with this disease. METHODS: The subjects were 66 patients with pancreatic head cancer who had undergone EUS. We examined each EUS finding as a possible prognostic predictor, including heterogeneity of internal echo, irregularity of peripheral echo, clarity of boundary echo, dilatation of the main pancreatic duct (MPD), dilatation of the common bile duct, lymph node swelling, vessel invasion, and the presence of ascites, by univariate and multivariate analysis for survival. RESULTS: Irregular peripheral echo, portal vein invasion, superior mesenteric artery/celiac artery invasion, and the presence of ascites were significant predictors of a poorer prognosis by univariate analysis for survival. In resectable cases, EUS findings of MPD dilatation and portal invasion were significant prognostic predictors by univariate analysis, and MPD dilatation was an independent prognostic predictor by multivariate analysis. CONCLUSION: EUS may be useful for predicting the prognosis of patients with pancreatic head cancer, based on the accuracy it provides in evaluating locoregional spreading. PMID- 15782095 TI - A pilot study of individualized maximum repeatable dose (iMRD), a new dose finding system, of weekly gemcitabine for patients with metastatic pancreas cancer. AB - OBJECTIVES: We developed and established a new dose-finding system, the individualized maximum repeatable dose (iMRD), suitable to induce prolonged TTP rather than tumor shrinkage. METHODS: We applied this system in weekly gemcitabine therapy for 18 metastatic pancreas cancer patients. We determined the iMRD at the 5th week, after weekly dose adjustments. We started at 500 mg/m2 (1/2 maximum tolerated dose) of gemcitabine and repeated the treatment with an increase or a decrease of 100 mg/m each week, if toxicity was 0 or more than grade 1, respectively. RESULTS: The iMRD of weekly gemcitabine was 300 mg/m2 in 2 patients, 400 mg/m2 in 3 patients, 500 mg/m2 in 5 patients, 600 mg/m2 in 6 patients, and 700 mg/m2 in 2 patients, demonstrating significant differences among individual patients. Grade 3 marrow depression occurred in only 1 patient (5.6%). Of these 18 patients, 3 (16.7%), 13 (72.2%) and 2 (11.1%) patients showed partial response, stable disease, and progressive disease, respectively. The median of times to progressive disease and survival were 4.5 and 9.5 months, respectively. There were no significant differences in 1-year survival time and more than 50% reduction rate of serum CA19-9, a tumor marker for pancreatic cancer, between patients with lower (500 mg/m2 or less) and higher (600 mg/m2 or more) iMRD. CONCLUSION: These results suggest that iMRD is a simple method to determine an individual's tailored dose for chemotherapy and could be the optimal dose for patients with noncurable cancers such as metastatic pancreas cancer. PMID- 15782096 TI - Surgical anatomy of the innervation of the major duodenal papilla in human and Suncus murinus, from the perspective of preserving innervation in organ-saving procedures. AB - OBJECTIVES: Few studies have focused on the detailed surgical anatomy of the innervation of the major duodenal papillary region, especially in relation to duodenum-preserving pancreatic head resection (DPPHR) and its modified procedures, which is crucial to preserving the innervation of the papillary region. The aim of this study is to clarify the neural distribution of the major duodenal papilla in humans. METHODS: The pancreas, duodenum, and surrounding structures were dissected in 10 cadavers and immersed in a 0.001% solution of alizarin red S in ethanol to stain the peripheral nerves. The details of the innervation in the above areas were confirmed using a binocular microscope. Similarly, the distribution in 10 Suncus murinus was examined by whole mount immunohistochemistry method with antineurofilament protein antibody. RESULTS: The innervation of the papillary region in humans involved 2 systems. One arose from the celiac plexus, which through the anterior hepatic plexus running along the arcades of the superior pancreaticoduodenal arteries and through the posterior hepatic plexus running along or accompanying the common bile duct (CBD) or Wirsung's duct, innervated the papillary region. The other arose from the superior mesenteric plexus wound around the arcades of the inferior pancreaticoduodenal arteries innervating the papillary region. The results in S. murinus supported those in humans. CONCLUSIONS: We emphasize the importance of the nervus-preserving of the major duodenal papilla and CBD by a suitable pancreatic head remnant, preserving the pancreaticoduodenal arterial arcades and avoiding kocherization of the CBD in DPPHR and its modified procedures. PMID- 15782097 TI - Renal cancer cell metastasis into the pancreas: a single-center experience and overview of the literature. AB - OBJECTIVES: The pancreas is a rare target for metastasis from other primary cancers, but pancreatic metastasis play a role in the diagnostic workup of patients with pancreatic tumors, especially in patients with a history of renal cell carcinoma (RCC). METHODS: Between October 2001 and June 2004 data from 601 patients undergoing pancreatic resection were entered prospectively in a database and were analyzed for metastasis into the pancreas from RCC. RESULTS: Fifteen patients with metastasis to the pancreas from RCC were identified. One patient showed metastatic disease at time of primary diagnosis. In 8 patients, the pancreas was the only site of metastasis, whereas in 7 patients, other organs, such as the thyroid gland, the lung, or the liver, were targets of metastasis, either metachronous or simultaneous at the time of pancreatic metastasis. The median interval between primary treatment of RCC and occurrence of pancreatic metastasis was 86 months (range, 0-258). Most patients were asymptomatic and diagnosed during standard tumor follow-up. So far, 14 patients remain alive with a median follow-up of 10 months. CONCLUSION: Pancreatic metastasis from RCC is rare but can occur even more than 20 years after primary tumor manifestation. Our results show that pancreatic resections for metastasis can be performed safely with a low rate of complications. Patients with a history of RCC should undergo a long-term follow-up to detect and evaluate for pancreatic metastases as well for metastasis to other organ sites. PMID- 15782098 TI - 5-fluorouracil intra-arterial infusion combined with systemic gemcitabine for unresectable pancreatic cancer. AB - OBJECTIVES: The aim of this study was to define assessment of response and adverse events of the combination chemotherapy of 5-fluorouracil (5-FU) pancreatic and hepatic arterial continuous infusion and systemic gemcitabine administration for unresectable pancreatic cancer. METHODS: We treated 24 chemotherapy-naive patients with unresectable pancreatic cancer. To prevent gastroduodenal injury from 5-FU infusion, the catheter was placed to allow the distribution of 5-FU to the pancreatic tumor and the liver after occlusion of the gastric and pancreaticoduodenal arteries. 5-FU was administered at a dose of 250 mg/d on days 1 to 5 every week as a continuous arterial infusion. Gemcitabine was infused intravenously at a dose of 1000 mg once weekly for 3 consecutive weeks of every 4 weeks. RESULTS: The partial response rate was 20.8% (5 of 24), although there was no case of complete response. Fourteen cases (58.3%) were stable disease, and 5 cases (20.8%) were progressive disease. The most common toxicities were hematological and gastrointestinal events. No patients died of adverse effects using this chemotherapy. Gastric and/or duodenal ulcers occurred because of 5-FU intra-arterial infusion. Catheter-related cholangitis occurred in patients with biliary drainage for obstructive jaundice. Median survival time was 14 months, with a 50.9% 1-year survival rate, although patients with performance status 2 and multiple organ metastases had a poor prognosis. CONCLUSIONS: This combination chemotherapy was well tolerated and seemed to be effective for patients with unresectable pancreatic cancer. PMID- 15782099 TI - Similarities of K+ATP channel expression and Ca2+ changes in pancreatic beta cells and hypothalamic neurons. AB - The mechanisms through which pancreatic beta cells recognize and respond to changes in circulating glucose are well understood. Evidence is accumulating that a subpopulation of neurons in the ventromedial hypothalamus (VMH) use similar cellular mechanisms to sense changes in extracellular glucose. In the present study, we used PCR and single-cell calcium imaging techniques to investigate whether glucose-sensing cells in the pancreas and hypothalamus employ a similar set of stimulus-response elements. Dispersed cells from mouse pancreata and hypothalamus were used in conjunction with the insulin-secreting cell line MIN6. We present functional data suggesting that both pancreatic and a subpopulation of hypothalamic cells exhibit glucose- and tolbutamide-evoked changes in cytosolic calcium and consider some clinical implications of different glucose sensors using the same mechanisms. PMID- 15782100 TI - Aberrant expression of cdx2 homeobox gene in intraductal papillary-mucinous neoplasm of the pancreas but not in pancreatic ductal adenocarcinoma. AB - OBJECTIVES: The authors investigated differential expression of cdx2 in intraductal papillary mucinous neoplasm of the pancreas (IPMN) and ductal adenocarcinoma. METHODS: Patients with IPMN (n = 23) and pancreatic ductal adenocarcinoma (n = 30) were studied. Expression of cdx2 mRNA, CDX2 protein, muc2 mRNA, MUC2 apomucin, and MUC5AC apomucin were determined. RESULTS: Of 23 IPMN, expression of cdx2 was detected in 8 of 9 adenomas, 6 of 8 borderline malignancies, and 3 of 6 malignancies, respectively. Of the 3 cdx2-expressing malignant IPMN, 2 did not display CDX2 expression. The appearance of CDX2 positive neoplastic cells in IPMN coincided with that of MUC2-positive neoplastic cells. All 23 IPMN displayed a positive MUC5AC expression irrespective to the status of CDX2 expression. Of 30 ductal adenocarcinomas, none displayed cdx2, CDX2, and MUC2 expression, while 10 cases displayed positive MUC5AC expression. CONCLUSION: Aberrant expression of cdx2 homeobox gene was detected in IPMN but not in ductal adenocarcinoma, which might determine their diverse carcinogenesis. PMID- 15782101 TI - Pancreatic secretory trypsin inhibitor (SPINK1) gene mutations in patients with acute pancreatitis. AB - OBJECTIVES: Mutations in the secretory trypsin inhibitor (SPINK1) gene have been found to be associated with hereditary and chronic pancreatitis. There are no previous reports on SPINK1 mutations in patients with acute pancreatitis (AP). METHODS: The study population consists of 371 patients with AP, of which 207 patients had mild and 164 had a severe form of the disease. The etiologies of AP were identified. Four hundred fifty-nine blood donors served as controls. SPINK1 N34S and P55S mutations were detected by minisequencing and confirmed by direct sequencing. RESULTS: The N34S mutation was found in 29 (7.8%) of the patients and in 12 (2.6%) of the controls (P < 0.0001, Fisher exact test). There was no difference in the frequency of the P55SS mutation between the groups. A majority of the patients (n = 229; 61.7%) had alcohol-induced AP. The frequency of the N34S mutation was higher in the subgroups of severe AP (15/164; 9.1%) and alcohol induced AP (21/229; 9.2%), but the differences were not statistically significant. No differences in age at admission and number of attacks of AP were observed between the groups. CONCLUSION: SPINK1 N34S mutation enhances the susceptibility of AP. PMID- 15782102 TI - Rapid urinary trypsinogen-2 test strip in the diagnosis of acute pancreatitis. AB - OBJECTIVES: Early diagnosis of acute pancreatitis remains a challenge. A rapid dipstick screening test for acute pancreatitis has been developed. This prospective study was designed to evaluate the diagnostic value and time course of the rapid urinary trypsinogen-2 test strip in acute pancreatitis, with comparisons with serum amylase and serum lipase. METHODS: A total of 165 patients with acute abdominal pain (67 with acute pancreatitis and 98 with other acute abdominal diseases) attending our emergency unit were included. All patients were tested with the urinary trypsinogen-2 test strip, and serum amylase and serum lipase concentrations were determined simultaneously. To measure the time course of the urinary trypsinogen-2 test, 32 patients with acute pancreatitis were tested with a urinary trypsinogen-2 test strip on days 1, 2, 3, and 4 after admission. RESULTS: Using a cutoff level of 50 microg/L for urinary trypsinogen 2, the sensitivity, specificity, and accuracy of the urinary trypsinogen-2 test strip for recognition of acute pancreatitis were 89.6%, 85.7%, and 87.3%, respectively. The diagnostic accuracy rates of serum amylase and serum lipase were 88.5% and 93.3%, using cutoff values of 3 times the upper normal limits for serum amylase and serum lipase, respectively. All but one of the 17 patients with severe acute pancreatitis was detected by the test strip (sensitivity, 94.1%). The time-course study of the urinary trypsinogen-2 test strip revealed that the sensitivity on days 1, 2, 3, and 4 was 90.6%, 81.2%, 59.4%, and 50%, respectively. There was no significant difference in the sensitivity between urinary trypsinogen-2 and serum lipase; however, the sensitivity values of serum lipase were significantly higher than those of serum amylase from days 1 to 4. CONCLUSION: The rapid urinary trypsinogen-2 test is a reliable and simple method for the early diagnosis of acute pancreatitis. A positive test identifies patients in need of further diagnostic measures. The urinary trypsinogen-2 test can be performed in health care units where laboratory testing facilities are not immediately available. PMID- 15782103 TI - Rat experimental model of continuous regional arterial infusion of protease inhibitor and its effects on severe acute pancreatitis. AB - OBJECTIVES: The rat experimental model of continuous regional arterial infusion of protease inhibitor (CRAI) on acute pancreatitis has yet to be established. Therefore, the aims of this study were (1) to establish the rat experimental model of CRAI and (2) to evaluate the effects of nafamostat on rat severe acute pancreatitis via different routes of administration. METHODS: The rat internal jugular vein or the celiac artery was infused with nafamostat, and the concentration of nafamostat in the lung and pancreas was measured. After the induction of severe acute pancreatitis, rats received intravenous or regional intraarterial infusion of nafamostat and then concentrations of trypsinogen activated peptide (TAP) and serum interleukin (IL-6), and histologic sections of the pancreas were examined and the 96-hour survival rate was evaluated. RESULTS: CRAI rats had higher concentrations of nafamostat in the pancreas than those infused intravenously. However, CRAI rats had lower concentrations of nafamostat in the lung that those infused intravenously. CRAI significantly reduced the levels of TAP and pancreatic necrosis. Moreover, the levels of serum IL-6 and the mortality rate were significantly reduced after CRAI compared with the intravenous infusion of nafamostat. CONCLUSION: The effectiveness of the rat experimental model of CRAI on acute pancreatitis was clearly demonstrated. The concentration of nafamostat in the lung and pancreas and the effects of nafamostat differ according to the route of administration. PMID- 15782104 TI - Microcirculatory function and tissue damage is improved after therapeutic injection of bovine hemoglobin in severe acute rodent pancreatitis. AB - OBJECTIVES: Stasis of the pancreatic microcirculation initiates and aggravates acute pancreatitis. Bovine hemoglobin has been shown to improve microcirculation in acute pancreatitis if prophylactically infused 15 minutes after initiation of acute pancreatitis. The purpose of this study was to evaluate the therapeutic effectiveness of bovine hemoglobin on pancreatic microcirculation and tissue damage later in the course of experimental acute rodent pancreatitis. METHODS: In Wistar rats, severe acute pancreatitis was induced by administration of glyco deoxycholic-acid intraductally and cerulein intravenously. Pancreatic microcirculation was continuously monitored by intravital microscopy. Three hours after the initiation of acute pancreatitis, animals received either 0.8 mL bovine hemoglobin (Oxyglobin), hydroxyethyl starch (HES), or 2.4 mL 0.9% NaCl intravenously at random. After 6 hours, animals were killed, and histopathological damage of the pancreas was assessed using a validated histology score. RESULTS: Pancreatic microcirculation assessed by leukocyte adherence was significantly improved by the administration of bovine hemoglobin in comparison with normal saline over time (mean difference, 51.6 +/- 9.2; P < 0.001) and HES (mean difference, 24.1 +/- 9.2; P = 0.037). This result was paralleled by decreased tissue damage in the bovine hemoglobin group as opposed to NaCl (6.75 vs. 12; range, 5.25-7.75 vs. 8.25-14; P < 0.001) and HES (6.75 vs. 9; range, 5.25 7.75 vs. 7.5-10.75; P < 0.001). CONCLUSION: Therapeutic intravenous infusion of bovine hemoglobin improves pancreatic microcirculation and reduces pancreatic tissue damage in severe acute rodent pancreatitis but is not as effective as early (prophylactic) administration. PMID- 15782105 TI - Transient receptor potential vanilloid (TRPV-1) promotes neurogenic inflammation in the pancreas via activation of the neurokinin-1 receptor (NK-1R). AB - OBJECTIVES: The transient receptor potential vanilloid 1 (TRPV-1) is an ion channel found on primary sensory afferent neurons. Activation of TRPV-1 leads to the release of the proinflammatory neuropeptide substance P (SP). SP then binds to the neurokinin-1 receptor (NK1-R) on endothelial cells and promotes extravasation of plasma and proteins into the interstitial tissue and neutrophil infiltration, a process called neurogenic inflammation. We tested 2 hypotheses: (1) activation of TRPV-1 in the pancreas leads to interstitial edema and neutrophil infiltration and (2) TRPV-1-induced plasma extravasation is mediated by the release of SP and activation of the NK1-R in the rat. METHODS: We measured extravasation of the intravascular tracer Evans blue as an index of plasma extravasation and quantified pancreas tissue myeloperoxidase activity (MPO) as a marker of neutrophil infiltration. The severity of inflammation following intravenous infusion of the secretagogue cerulein (10 microg/kg/h x 4 hours) was assessed using a histologic scoring system. RESULTS: Intravenous injection of the TRPV-1 agonist capsaicin induced a dose-dependent increase in Evans blue accumulation in the rat pancreas (P < 0.05 vs. vehicle control). This effect was blocked by pretreatment with the TRPV-1 antagonist capsazepine (1.8 mg/kg), or the NK1-R antagonist CP 96,345 (1 mg/kg). Capsazepine also reduced cerulein induced Evans blue, MPO, and histologic severity of inflammation in the pancreas but had no effect on serum amylase. CONCLUSION: Activation of TRPV-1 induces SP mediated plasma extravasation in the rat pancreas via activation of the NK1-R. TRPV-1 mediates neurogenic inflammation in cerulein-induced pancreatitis in the rat. PMID- 15782106 TI - Development and validation of an experimental model for the assessment of radiofrequency ablation of pancreatic parenchyma. AB - OBJECTIVES: The aim of this study was to develop and validate an ex vivo model for the assessment of radiofrequency (RF) ablation of the pancreas. METHODS: Porcine pancreata were used within 1 hour of sacrifice. RF was delivered to a premarked area in the center of the pancreatic head using a thermocouple-tipped multiprobe array. Four temperature presets were evaluated: 70 degrees-100 degrees C in 10 degrees increments. Immediately after ablation serial sections of the pancreatic head were cut to incorporate duodenum, portal vein, and bile duct. For each experiment, a portion of pancreatic tail was sampled as nonablated control. Hematoxylin and eosin (H&E) slides together with nicotinamide adenine dinucleotide (NADH) stained preparations were made. The NADH staining was quantified using computerized digital image recognition techniques. RESULTS: Control sections (n = 20) demonstrated normal pancreatic architecture on H&E and strong NADH staining indicating preserved tissue oxidative metabolism. RF produced a temperature-dependent destruction of parenchymal architecture (H&E) with a corresponding loss of NADH activity. There was no evidence of thermal injury to the duodenum. Quantification of NADH staining demonstrated a median positive staining of 69.26% (55.87-97.28) for control tissue compared with 1.40% (0-7.77) for ablated pancreas (P < 0.001; Mann-Whitney U test). CONCLUSION: This study describes the development of a relatively simple, reliable, and reproducible model for evaluation of RF ablation of pancreatic parenchyma. PMID- 15782107 TI - Comparison of histology and extracellular matrix between autoimmune and alcoholic chronic pancreatitis. AB - OBJECTIVES: In autoimmune chronic pancreatitis (AIP), the histology is known to be characteristic and histologic recovery including pancreatic fibrosis has been reported after steroid therapy. The aims of this study were to demonstrate whether these histologic findings were unique to AIP and evaluate any differences in the composition of extracellular matrix between AIP and ordinary chronic pancreatitis. METHODS: Histologic findings and extracellular matrix proteins (collagen types I, III, and IV and fibronectin) were evaluated in 15 patients with clinically proven AIP and compared with those of 8 patients with surgically treated alcoholic chronic pancreatitis (ACP). RESULTS: The pattern of fibrosis was mainly loose fibrosis with stromal edema in AIP, while it was dense fibrosis in ACP. Acinar atrophy was more diffuse and severe in AIP than ACP. While diffuse with same stage of inflammatory activity was observed in AIP, multifocal inflammation with different stage of inflammatory was seen in ACP. For extracellular matrix proteins, dense deposition (compared with normal controls) of collagen type III was observed more frequently in ACP than AIP (P < 0.05). ACP showed decreased deposition of collagen type IV more frequently than AIP (P = 0.001). CONCLUSION: Fibrosis and inflammation are common to both AIP and ACP, but the pattern is very different between the 2 groups, and the deposition of collagen types III and IV is substantially different between AIP and ACP. PMID- 15782108 TI - Use of recombinant activated factor VII for bleeding in pancreatitis: a case series. AB - OBJECTIVES: To describe the effects of recombinant activated factor VII (rFVIIa) in the treatment of bleeding in a series of patients with acute or chronic pancreatitis. METHODS: Twelve patients (age, 2.5-65 years) with pancreatitis and bleeding were treated with 18.5 to 120 microg/kg of rFVIIa. Eight patients also had sepsis/infection and/or disseminated intravascular coagulation (DIC). The effects of rFVIIa on bleeding, coagulation status, and transfusion requirements were noted. RESULTS: Bleeding stopped in 4 patients, was markedly reduced in 4 patients, was reduced in 3 patients, and was remained unchanged in 1 patient. For most patients with pre- and post-rFVIIa data, coagulation parameters improved and transfusion requirements reduced. No thrombotic adverse events occurred. Seven patients died for reasons considered to be unrelated to rFVIIa treatment. CONCLUSIONS: This case series indicates that rFVIIa may be an effective hemostatic treatment of patients with pancreatitis suffering from massive bleeding. There were no thromboembolic events in any patient, including those with sepsis or DIC. PMID- 15782109 TI - Encephalopathy and acute axonal sensorimotor polyneuropathy following acute pancreatitis: a case report and review of the literature. AB - A case of a 22-year-old woman with rare neurologic complications including encephalopathy and acute axonal sensorimotor polyneuropathy in the course of acute pancreatitis is reported. The encephalopathy emerged 3 weeks after the onset of the illness with complete remission being noted 1 week later. The polyneuropathy presented as quadriplegia and respiratory failure that required intubation and partially remitted gradually. There was no pancreatic lesion, no major pancreatic surgery, no sepsis, and no multiple organ failure, all of which had been proposed as the predisposing factors. Severe inflammatory response syndrome (SIRS) that developed during the clinical course of this patient might have induced these neurologic complications. PMID- 15782110 TI - The first initiative targeted to increase the training of African-American scientists in pancreatic cancer research: the Mayo Clinic College of Medicine Oakwood College alliance. PMID- 15782111 TI - G-protein-activated phospholipase C-beta, new partners for cell polarity proteins Par3 and Par6. AB - Cell polarity and asymmetric cell division are fundamental traits of all living cells and play an essential role in embryonic development, neuronal cell chirality formation, and maintenance of mammalian epithelial cell morphology. Heterotrimeric GTP-binding proteins (G proteins) are involved in directing cell polarity and asymmetric cell division in different organisms. However, the mechanism for G-protein-mediated cell polarity and asymmetric cell division is poorly understood. In this study, we have demonstrated that G-protein-activated phospholipase C-beta (PLC-beta) interacts with cell polarity proteins Par3 and Par6 (Par: partition-defective) to form protein complexes and to mediate downstream signal transduction. The interactions between PLC-beta and Par proteins are direct and require the extreme C-terminal-specific sequence motifs of PLC-beta and the PDZ (PSD95/Dlg/ZO-1) domains of Par proteins. Binding of Par proteins with PLC-beta stimulates PLC-beta enzymatic activity, leading to the hydrolysis of phosphatidylinositol-4,5-bisphosphate, and the production of diacylglycerol and inositol 1,4,5-triphosphate, important mediators in cell polarity and cell asymmetric division processes. Furthermore, we have shown that coexpression of PLC-beta with Par proteins induces transcriptional activation coupled to intracellular Ca2+ and the Wnt signaling pathway. Therefore, our data suggest that the interaction of PLC-beta with cell polarity Par proteins may serve as a nexus to transduce extracellular signals to transcriptional regulation through G-protein-mediated signaling pathway in cell polarity and cell asymmetric division. PMID- 15782112 TI - Wild-type p53 stimulates homologous recombination upon sequence-specific binding to the ribosomal gene cluster repeat. AB - p53 plays a central role in the maintenance of the genome integrity, both as a gatekeeper and a caretaker. Sequence-specific recognition of DNA is underlying the ability of p53 to transcriptionally transactivate target genes during checkpoint control and to regulate DNA replication at the TGCCT repeat from the ribosomal gene cluster (RGC). In contrast, suppression of recombination by p53 has been observed with nonconsensus DNA sequences. In this study, we discovered that wild-type p53 stimulates homologous recombination adjacent to the RGC repeat, whereas downregulation is seen with a mutated version thereof and with a microsatellite repeat sequence. Analysis of the causes possibly underlying the enhancement of homologous recombination revealed that p53 binding to the RGC element delays DNA synthesis. This was demonstrated after integration of the corresponding DNA fragments into our Simian virus 40-based model system, which was used to study recombination on replicating minichromosomes. Differently, with plasmid-based substrates, p53 did not stimulate recombination at the RGC sequence. Thus, in combination with our previous findings, p53 may promote homologous recombination by two separate mechanisms involving either molecular interactions with topoisomerase I or/and by specific binding to certain genomic regions, thereby causing replication fork stalling and recombination. PMID- 15782113 TI - Partial downregulation of MAD1 causes spindle checkpoint inactivation and aneuploidy, but does not confer resistance towards taxol. AB - The mitotic spindle assembly checkpoint ensures proper chromosome segregation during mitosis by inhibiting the onset of anaphase until all kinetochores are attached to the mitotic spindle and tension across the kinetochores is generated. Here, we report that the stable partial downregulation of the spindle checkpoint gene MAD1, which is observed in human cancer, leads to a functional inactivation of the spindle checkpoint resulting in gross aneuploidy. Interestingly, although Mad1 is thought to act as a kinetochore based activator of Mad2 during checkpoint activation, we show that normal levels of Mad2, but not of Mad1, are required for preventing premature sister chromatid separation and for maintaining the timing of an undisturbed mitosis, suggesting a Mad1 independent function of Mad2 that operates independent of its checkpoint function. Most significantly, a partial repression of either MAD1 or MAD2 confers resistance to nocodazole, a drug that inhibits microtubule attachment. In contrast, sensitivity to clinically relevant drugs like taxol or monastrol that inhibit the generation of tension across kinetochores is not modulated by partial downregulation of MAD1, suggesting a functional bifurcation of spindle checkpoint dependent apoptotic pathways. PMID- 15782114 TI - The EphA8 receptor induces sustained MAP kinase activation to promote neurite outgrowth in neuronal cells. AB - Recent studies in our laboratory demonstrate that ligand-mediated activation of the EphA8 receptor critically regulates cell adhesion and migration. In this report, we show that the EphA8 receptor induces neurite outgrowth in NG108-15 cells in the absence of ligand stimulation. Using various deletion mutants lacking specific intracytoplasmic regions, we confirm that the tyrosine kinase domain of EphA8 is important for inducing neurite outgrowth. However, the tyrosine kinase activity of EphA8 is not crucial for neurite outgrowth induction. Treatment with various inhibitors further reveals that the mitogen-activated protein kinase (MAPK) signaling pathway is critical for neurite outgrowth induced by EphA8. Consistent with these results, EphA8 expression induced a sustained increase in the activity of MAPK, whereas ligand-mediated EphA8 activation had no further modulatory effects on MAP kinase activity. Additionally, activated MAPK relocalized from the cytoplasm to the nucleus in response to EphA8 transfection. These results collectively suggest that the EphA8 receptor is capable of inducing a sustained increase in MAPK activity, thereby promoting neurite outgrowth in neuronal cells. PMID- 15782115 TI - Hypersensitivity of Brca1-deficient MEF to the DNA interstrand crosslinking agent mitomycin C is associated with defect in homologous recombination repair and aberrant S-phase arrest. AB - Hypersensitivity of Brca1-deficient cells to interstrand crosslinking (ICL) agents such as cisplatin and mitomycin C (MMC) implicates an important role for Brca1 in cellular response to the ICL DNA damage repair. However, the detailed mechanism of how Brca1 is involved in the ICL response remains unclear. In this study, we analysed the cellular response to MMC treatment using isogenic mouse embryonic fibroblasts (MEFs) including wild type, p53-/- and p53-/-Brca1-/-. Marked hypersensitivity of p53-/- Brca1-/- MEFs to MMC was found, and the reconstitution of Brca1 expression in these cells restored resistance to MMC. Upon MMC treatment, wild-type MEF was temporarily arrested at G2/M phase but subsequently resumed a normal cell cycle progression. In contrast, Brca1 deficient MEF exhibited a marked time-dependent accumulation of cells arrested at S phase and a prolonged increase in the G2/M population, followed by extensive cell deaths. Importantly, DNA damage-induced Rad51 foci were not formed in these cells, suggesting a defect in homologous recombination. Such defects are fully rescued by reconstitution of Brca1 expression in Brca1-deficient MEF, suggesting that Brca1 directly plays an essential role in ICL repair, which depends on homologous recombination during S phase. PMID- 15782116 TI - Multimodality expression profiling shows SEPT9 to be overexpressed in a wide range of human tumours. AB - Septins are an evolutionarily conserved family of GTPases with diverse functions including roles in cytokinesis that have been implicated in neoplasia. To address the potential role of SEPT9 in tumorigenesis, we assessed the expression of SEPT9 in 7287 fresh frozen human tissue samples and 292 human cell lines by microarray analysis. In addition, we used a sensitive RT-PCR strategy to define the expression of SEPT9 isoforms in archival formalin-fixed and paraffin-embedded normal human tissues. The mRNA data were further confirmed by immunohistological analyses of SEPT9 protein expression in normal human tissues using antisera that detect SEPT9 isoforms. Using these complementary approaches, we demonstrate that SEPT9 mRNA and protein are expressed ubiquitously, with the isoforms showing tissue-specific expression. The microarray analysis indicates that there is consistent overexpression of SEPT9 in diverse human tumours including breast, CNS, endometrium, kidney, liver, lung, lymphoid, oesophagus, ovary, pancreas, skin, soft tissue and thyroid. Since tumours are commonly associated with enhanced cell proliferation, we examined the possible correlation of Ki67 and SEPT9 expression in normal tissues and tumours. Our data indicate that the overexpression of SEPT9 in neoplasia is not simply a proliferation-associated phenomenon, despite its role in cytokinesis. PMID- 15782118 TI - BRAF-V600E is not involved in the colorectal tumorigenesis of HNPCC in patients with functional MLH1 and MSH2 genes. AB - Recently, it was shown that the oncogenic activation of BRAF, a member of the RAS/RAF family of kinases, by the V600E mutation is characteristic for sporadic colon tumors with microsatellite instability. Further, it was shown to associate with the silencing of the mismatch repair (MMR) gene MLH1 by hypermethylation. Moreover, BRAF mutations proved to be absent in tumors from hereditary nonpolyposis colorectal cancer syndrome (HNPCC) families with germline mutations in the MMR genes MLH1 and MSH2. These data suggest that the oncogenic activation of BRAF is involved only in sporadic colorectal tumorigenesis. In order to further support this hypothesis, we have extended the analysis of the BRAF gene to a different subset of HNPCC families without germline mutations in MLH1 and MSH2. BRAF-V600E mutations were analysed by automatic sequencing in 38 tumors from HNPCC families with germline mutations in the MSH6 gene and also in HNPCC (suspected) families that do not have mutations in the MMR genes MLH1, MSH2 and MSH6. All patients belong to different families. No mutations were detected in 14 tumors from HNPCC patients with germline mutations in MSH6. Further, no mutations of BRAF were found in tumors from 23 MMR-negative families, from which 13 fulfilled the Amsterdam criteria (HNPCC) and 10 were suspected for HNPCC as they were positive for the Bethesda criteria. Overall, our data reinforce the concept that BRAF is not involved in the colorectal tumorigenesis of HNPCC. The detection of a positive BRAF-V600E mutation in a colorectal cancer suggests a sporadic origin of the disease and the absence of germline alterations of MLH1, MSH2 and also of MSH6. These findings have a potential impact in the genetic testing for HNPCC diagnostics and suggest a potential use of BRAF as exclusion criteria for HNPCC or as a molecular marker of sporadic cancer. PMID- 15782117 TI - The transcriptional factor YY1 upregulates the novel invasion suppressor HLJ1 expression and inhibits cancer cell invasion. AB - By using microarray and an invasion/metastasis lung cell line model, we identified the DnaJ-like heat shock protein 40, HLJ1, and found that the expression of HLJ1 correlates negatively with cancer cell invasion ability. Overexpression of HLJ1 can suppress cancer cell invasion in vitro. We further characterize the putative promoter region and investigate the transcriptional regulations of human HLJ1. A serial deletion of the 1.2 kb at the 5'-flanking region of the human HLJ1 gene was subcloned into a vector containing reporter gene and transfected into human lung adenocarcinoma cell line CL1-0, followed by luciferase activity assay. The results indicated that the region from -232 to +176 could drive the basal transcriptional activity of the HLJ1 gene. Sequence analysis of the HLJ1 gene promoter region showed absence of a TATA box, but identified an inverted CCAAT box and four YY1 transcriptional factor-binding sites, which may be important in the regulation of HLJ1 expression. Co transfection of the YY1 and HLJ1 basal promoter regions, site-directed mutagenesis, and electrophoretic mobility shift assay confirmed that YY1 could upregulate HLJ1 basal promoter activity. Furthermore, we also demonstrated that overexpression of YY1 in CL1-0 cells can increase HLJ1 expression and reduce cell invasive capability. The reduction of cancer cell invasive ability is, at least in part, through upregulation of E-cadherin expression. The increase in HLJ1 and E-cadherin expression, as well as the suppression of invasion ability, can be reversed specifically by HLJ1 siRNA. PMID- 15782119 TI - Aberrant NF-kappaB2/p52 expression in Hodgkin/Reed-Sternberg cells and CD30 transformed rat fibroblasts. AB - Overexpression of CD30 and constitutive nuclear factor-kappaB (NF-kappaB) activation are hallmarks of the malignant Hodgkin Reed-Sternberg (H-RS) cells. Previous investigations have demonstrated that both proliferation and survival of H-RS cells require constitutive NF-kappaB activity, which is comprised of the p50 and RelA subunits. We report here enhanced expression of NF-kappaB2/p52 and RelB containing NF-kappaB DNA-binding activity in Epstein-Barr virus-negative H-RS cells. Kinetic studies revealed that a proteasome inhibitor MG132 induced p100 accumulation with reduced p52 expression in H-RS cells, suggesting proteasome dependent processing of p100. In addition, treatment with a protein synthesis inhibitor cycloheximide rapidly downregulated inhibitor of NF-kappaB (IkappaB) kinase activity in H-RS cells. We also demonstrate that overexpression of CD30 in rat fibroblasts at levels comparable to those in H-RS cells results in constitutive IkappaB kinase activation, proteasome-dependent p100 processing, and NF-kappaB-dependent cell transformation. Our results thus indicate that CD30 triggers the noncanonical NF-kappaB activation pathway, and suggest that deregulated CD30 signaling contributes to the neoplastic features of H-RS cells. PMID- 15782120 TI - Dinucleotide repeats negatively modulate the promoter activity of Cyr61 and is unstable in hepatocellular carcinoma patients. AB - Cyr61 is a secreted, cysteine-rich, heparin-binding protein that mediates diverse functions including extracellular matrix formation, differentiation, cell proliferation, adhesion, migration, survival, as well as angiogenesis and tumorigenesis. In this study, we found that Cyr61 gene expression is significantly downregulated in the tumors of hepatocellular carcinoma (HCC) patients. To elucidate its mechanism of gene regulation, we examined the promoter of Cyr61 which contains two long stretches of repeats, each comprising d(CA) dinucleotide repeats downstream of HNF3beta- and ATF-binding sites. We hypothesized that the d(CA) repeats may play an important role in regulating Cyr61 promoter activity and performed promoter reporter assays to examine this. We found that a greater number of d(CA) repeats resulted in significantly lower promoter activity of the Cyr61 gene in the KB3-1 and HepG2 cell lines, but not in the MCF-7 cell line. In addition, the d(CA) repeats, but not other random sequences, were found to be important for Cyr61 promoter activity. We further demonstrate that the ATF- and HNF3beta-binding sites upstream the d(CA) repeats positively and negatively modulate Cyr61 promoter activity, respectively. An examination of the d(CA) dinucleotide patterns in the Cyr61 promoter in HCC patients revealed that approximately 32% of these patients exhibited either loss of heterozygosity or somatic mosaicism in either the tumors, adjacent normal liver tissues or both. PMID- 15782121 TI - Hsp90-Akt phosphorylates ASK1 and inhibits ASK1-mediated apoptosis. AB - Hsp90 client protein Akt has been shown to inhibit cell apoptosis in part by inhibiting proapoptotic kinase ASK1 (apoptosis signal-regulating kinase 1) activity. In the present study, we show that Hsp90 inhibits hydrogen peroxide (H(2)O(2))-induced ASK1-p38 activation in endothelial cells (EC). The inhibitory effect of Hsp90 on ASK1-p38 activities is diminished when the Akt phosphorylation site on ASK1 (pSer83) is absent or when Akt is genetically deleted in cells, suggesting that Hsp90 and Akt function together to inhibit ASK1-p38 signaling. Thus, inhibition of Hsp90 by 17-allyamino-17-demethoxygeldanamycin (17-AAG) or phosphatidylinositol 3-kinase (PI3K) LY294002 induced and synergized ASK1 activation and ASK1-mediated EC apoptosis. Furthermore, we show that in resting EC Hsp90, Akt and ASK1 form a ternary complex in which both Akt and ASK1 bind to the middle domain of Hsp90, suggesting that Hsp90 may hold Akt and ASK1 in close proximity. The N-terminal domain of ASK1 containing the Akt phosphorylation site (pSer83) associates with Akt in resting state. However, Akt is released from the N-terminal domain concomitant with binding to the C-terminal domain of ASK1 in response to ASK1 activator H(2)O(2), inhibitor of Hsp90 17-AAG and Akt inhibitor LY294002, leading to a more stable Hsp90-Akt-ASK1 complex. We conclude that Hsp90 Akt forms a complex with ASK1 and protect EC from stress-induced apoptosis. PMID- 15782122 TI - Opposing effects of bovine papillomavirus type 1 E6 and E7 genes on Fas-mediated apoptosis. AB - Programmed cell death (PCD), best exemplified by apoptosis, is a genetically programmed process of cellular destruction that is indispensable for normal development and homeostasis of multicellular organisms. Tumor necrosis factor alpha (TNF) and related cytokines are employed by host defenses to eliminate virally infected cells through induction of apoptosis. Many viruses have evolved specific gene products to modulate this process. We have recently shown that the bovine papillomavirus type 1 (BPV-1) E6 and E7 genes independently sensitize mouse cells to TNF-induced apoptosis. In this report, we investigated the effect of E6 and E7 expression on Fas-mediated apoptosis. In contrast to TNF-mediated apoptosis, E6 and E7 demonstrated opposite effects: while E7 potentiated apoptosis triggered by an agonistic Fas antibody, E6 attenuated the effect. The mitochondrial pathway leading to the activation of caspases appears to be involved in Fas-mediated apoptosis in C127 cells. To further explore the mechanisms by which E6 and E7 modulate Fas-mediated apoptosis, we examined the surface expression of Fas in cells expressing E6 and E7. Significantly, levels of surface Fas expression correlated with the opposing effects of E6 and E7 on Fas mediated apoptosis. Specifically, while E7 increased the surface expression of Fas, E6 reduced surface Fas expression. Mutational analysis demonstrated a correlation of E6's ability to downregulate surface Fas expression and apoptosis. Since the tumor suppressor p53 can be targeted for degradation by human papillomavirus and has been shown to induce apoptosis by upregulating surface Fas expression, we investigated the role of p53 in BPV-1 E6 and E7 modulation of Fas mediated apoptosis. Our results demonstrated that the modulatory effects by E6 and E7 could occur in the absence of p53. Interestingly, the reduced Fas protein level on the cell surface is not accompanied by a decrease in total Fas levels in E6-expressing cells. Instead, considerably more Fas protein is found in the cytoplasm of cells expressing E6. These results highlight a novel activity of E6 and E7 that may be involved in viral pathogenesis. PMID- 15782124 TI - Impaired homologous recombination DNA repair and enhanced sensitivity to DNA damage in prostate cancer cells exposed to anchorage-independence. AB - During metastases, cancer cells are temporarily exposed to the condition in which interactions with extracellular environment can be restricted (anchorage independence). We demonstrate that the sensitivity of prostate cancer cell lines, DU145 and PC-3, to genotoxic treatment (cisplatin and gamma-irradiation) increased several folds when cells were forced to grow in anchorage-independence. This enhanced drug sensitivity was associated with a severe impairment of homologous recombination-directed DNA repair (HRR). The mechanism involves Rad51, which is the major enzymatic component of HRR. The protein level of Rad51 and its recruitment to DNA double-strand breaks (DSBs) were both attenuated. Rad51 deficiency in anchorage-independence was not associated with Rad51 promoter activity, and was not compensated by a constitutive overexpression of Rad51 cDNA. Instead, Rad51 protein level and its ability to colocalize with DSBs were restored in the presence of proteosome inhibitors, or when cells from the suspension cultures were allowed reattachment. Presented results indicate that anchorage-independence sensitizes prostate cancer cells to genotoxic agents; however, it also attenuates faithful component of DNA repair by targeting stability of Rad51. This temporal attenuation of HRR may contribute to the accumulation mutations after DNA damage, and possibly the selection of new adaptations in cells, which survived genotoxic treatment. PMID- 15782123 TI - p44/42 MAP kinase-dependent regulation of catalase by autocrine human growth hormone protects human mammary carcinoma cells from oxidative stress-induced apoptosis. AB - Previous microarray expression analyses have indicated autocrine human growth hormone (hGH) regulation of genes involved in the oxidative stress response. Expression analysis of antioxidant enzymes revealed that autocrine hGH increased both the mRNA and protein levels of catalase, superoxide dismutase 1 (SOD1), glutathione peroxidase and glutamylcysteine synthetase but not that of SOD2. As a consequence, autocrine hGH increased the antioxidant capacity of mammary carcinoma cells and protected against oxidative stress-induced apoptosis. Catalase activity was increased by autocrine production of hGH in mammary carcinoma cells and a catalase inhibitor abrogated protection from oxidative stress afforded by autocrine hGH. Autocrine hGH transcriptionally regulated catalase gene expression in a p44/42 MAP kinase-dependent manner and inhibition of MEK concordantly abrogated the protective effect of autocrine hGH against oxidative stress-induced apoptosis. Given that increased cellular oxidative stress is a key effector mechanism of specific chemotherapeutic agents, we propose that antagonism of autocrine hGH will improve the efficacy of chemotherapeutic regimes utilized for human mammary carcinoma. PMID- 15782125 TI - Mdm2 and mdmX prevent ASPP1 and ASPP2 from stimulating p53 without targeting p53 for degradation. AB - Using various mutants of p53 and mdm2, we demonstrate here that both the DNA binding and transactivation function of p53 are required for ASPP1 and ASPP2 to stimulate the apoptotic functions of p53. Mdm2 and mdmx prevent ASPP1 and ASPP2 from stimulating the apoptotic function of p53 by binding and inhibiting the transcriptional activity of p53. Importantly, mdm2 and mdmx can prevent the stimulatory effects of ASPP1 and ASPP2 without targeting p53 for degradation. These data provide a novel mechanism by which mdm2 and mdmx act as potent inhibitors of p53. PMID- 15782126 TI - Identification of proteolytic fragments from ErbB-2 that induce apoptosis. AB - The receptor tyrosine kinase ErbB-2 plays an important role in cell proliferation and differentiation as well as oncogenesis. We have found that ErbB-2 kinase domain fragmentation is important for the induction of apoptosis. Exogenous expression of peptides derived from the ErbB-2 kinase domain induces cells death with the hallmarks of apoptosis. In contrast, transfection of the ErbB-2 carboxy terminal domain did not induce apoptosis. We have identified a 37-residue segment from the ErbB-2 kinase N-terminal lobe that can strongly induce apoptosis in transfected cells. Cell death was not blocked by the pan-caspase inhibitor z-VAD FMK. Similar fragments derived from several other receptor tyrosine kinases also induce cell death. These data imply that proteolytic fragmentation of tyrosine kinases liberates apoptotic fragments that can accelerate cell death. PMID- 15782128 TI - Modulation of specific protein expression levels by PTEN: identification of AKAP121, DHFR, G3BP, Rap1, and RCC1 as potential targets of PTEN. AB - The tumor suppressor PTEN is mutated in a high percentage of human cancers, and is implicated in pathways regulating cell growth, proliferation, survival, and migration. Despite significant advances, our understanding of its mechanisms of action remains incomplete. We have used a high-throughput proteomic immunoblotting approach to identify proteins whose expression levels are modulated by PTEN. Out of over 800 proteins screened, 22 proteins showed significant changes in expression. Five proteins that exhibited two-fold or greater changes in expression level were further characterized. AKAP121 and G3BP expression was reduced, while dihydrofolate reductase (DHFR), Rap1 and RCC1 expression was elevated in response to PTEN expression in a PTEN-null T-cell leukemia line. The phosphatase activity of PTEN was required for these effects. However, direct inhibition of PI-3 Kinase could mimic PTEN in modulating expression of DHFR, G3BP, Rap1 and RCC1, but not AKAP121. Real-time PCR showed that the effects of PTEN were primarily post-transcriptional, and would not have been revealed by mRNA-based screens. We conclude from these data that PTEN can modulate the expression level of a number of different proteins. The identified proteins have the potential to serve as previously unrecognized effectors of PTEN, and suggest the existence of additional complexity in the modes by which PTEN can regulate cellular biology. PMID- 15782129 TI - Geldanamycins exquisitely inhibit HGF/SF-mediated tumor cell invasion. AB - Induction of the urokinase-type plasminogen activator (uPA) by hepatocyte growth factor/scatter factor (HGF/SF) plays an important role in tumor cell invasion and metastasis that is mediated through the Met receptor tyrosine kinase. Geldanamycins (GA) are antitumor drugs that bind and inhibit HSP90 chaperone activity at nanomolar concentrations (nM-GAi) by preventing proper folding and functioning of certain oncoproteins. Previously, we have shown that a subset of GA derivatives exhibit exquisite potency, inhibiting HGF/SF-induced uPA-plasmin activation at femtomolar concentrations (fM-GAi) in canine MDCK cells. Here, we report that (1) inhibition of HGF/SF-induced uPA activity by fM-GAi is not uncommon, in that several human tumor glioblastoma cell lines (DBTRG, U373 and SNB19), as well as SK-LMS-1 human leiomyosarcoma cells are also sensitive to fM GAi; (2) fM-GAi drugs only display inhibitory activity against HGF/SF-induced uPA activity (rather than basal activity), and only when the observed magnitude of uPA activity induction by HGF/SF is at least 1.5 times basal uPA activity; and (3) not only do fM-GAi derivatives strongly inhibit uPA activity but they also block MDCK cell scattering and in vitro invasion of human glioblastoma cells at similarly low drug concentrations. These effects of fM-GAi drugs on the Met activated signaling pathway occur at concentrations well below those required to measurably affect Met expression or cell proliferation. We also examined the effect of Radicicol (RA), a drug with higher affinity than GA for HSP90. RA displays uPA activity inhibition at nanomolar levels, but not at lower concentrations, indicating that HSP90 is not likely the fM-GAi molecular target. Thus, we show that certain GA drugs (fM-GAi) in an HGF/SF-dependent manner block uPA-plasmin activation in tumor cells at femtomolar levels. This inhibition can also be observed in scattering and in vitro invasion assays. Our findings also provide strong circumstantial evidence for a novel non-HSP90 molecular target that is involved in HGF/SF-mediated tumor cell invasion. PMID- 15782127 TI - 2-Methoxyestradiol-induced apoptosis in human leukemia cells proceeds through a reactive oxygen species and Akt-dependent process. AB - The effects of 2-Methoxyestradiol (2ME)-induced apoptosis was examined in human leukemia cells (U937 and Jurkat) in relation to mitochondrial injury, oxidative damage, and perturbations in signaling pathways. 2ME induced apoptosis in these cells in a dose-dependent manner associated with release of mitochondrial proteins (cytochrome c, AIF), generation of reactive oxygen species (ROS), downregulation of Mcl-1 and XIAP, and inactivation (dephosphorylation) of Akt accompanied by activation of JNK. In these cells, enforced activation of Akt by a constitutively active myristolated Akt construct prevented 2ME-mediated mitochondrial injury, XIAP and Mcl-1 downregulation, JNK activation, and apoptosis, but not ROS generation. Conversely, 2ME lethality was potentiated by the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002. Furthermore, in U937 cells, the hydrogen peroxide scavenger catalase and a superoxide dismutase (SOD) mimetic, TBAP, blocked these events, as well as Akt inactivation. Interruption of the JNK pathway by pharmacologic or genetic (e.g. siRNA) means attenuated 2ME induced mitochondrial injury, XIAP and Mcl-1 downregulation, and apoptosis. Collectively, these findings suggest a hierarchical model of 2ME-related apoptosis induction in human leukemia cells in which 2ME-induced oxidative injury represents a primary event resulting in Akt inactivation, leading, in turn, to JNK activation, and culminating in XIAP and Mcl-1 downregulation, mitochondrial injury, and apoptosis. They also suggest that in human leukemia cells, the Akt pathway plays a critical role in mediating the response to oxidative stress induced by 2ME. PMID- 15782130 TI - BARD1 induces apoptosis by catalysing phosphorylation of p53 by DNA-damage response kinase. AB - The BRCA1-associated RING domain protein BARD1 acts with BRCA1 in double-strand break repair and ubiquitination. BARD1 plays a role as mediator of apoptosis by binding to and stabilizing p53, and BARD1-repressed cells are resistant to apoptosis. We therefore investigated the mechanism by which BARD1 induces p53 stability and apoptosis. The apoptotic activity of p53 is regulated by phosphorylation. We demonstrate that BARD1 binds to unphosphorylated and serine 15 phosphorylated forms of p53 in several cell types and that the region required for binding comprises the region sufficient for apoptosis induction. In addition, BARD1 binds to Ku-70, the regulatory subunit of DNA-PK, suggesting that the mechanism of p53-induced apoptosis requires BARD1 for the phosphorylation of p53. Upregulation of BARD1 alone is sufficient for stabilization of p53 and phosphorylation on serine-15, as shown in nonmalignant epithelial cells and ovarian cancer cells, NuTu-19, which are defective in apoptosis induction and express aberrant splice variants of BARD1. Stabilization and phosphorylation of p53 in NuTu-19 cells, as well as apoptosis, can be induced by the exogenous expression of wild-type BARD1, suggesting that BARD1, by binding to the kinase and its substrate, catalyses p53 phosphorylation. PMID- 15782131 TI - Uncovering MYC's full oncogenic potential in the hematopoietic system. AB - MYC is an important oncogene in hematopoietic neoplasms in humans, yet the mechanism by which MYC induces the malignant transformation of blood cells has remained elusive. Postulating that mouse models of deregulated MYC expression may be helpful for advancing our understanding of MYC-induced hematopoietic malignancies, Suzanne Cory and her associates took advantage of the Vav promoter to express MYC throughout the hematopoietic system in transgenic mice. They report that the newly developed strain, referred to as VavP-MYC17, is prone to mature T-cell lymphomas (for which few good mouse models exist). They further show that VavP-MYC17 mice that are devoid of mature T cells (and B cells) because of genetic deficiency in Rag1 recombinase develop neoplasms of three distinct blood cell lineages: pre-T cells, pro-B cells, and macrophages. These findings establish that VavP-driven MYC has broad oncogenic potential in the hematopoietic compartment and prompts new views of the cellular assaults of deregulated MYC on hematopoietic stem and early progenitor cells. PMID- 15782132 TI - Rapamycin induces apoptosis of JN-DSRCT-1 cells by increasing the Bax : Bcl-xL ratio through concurrent mechanisms dependent and independent of its mTOR inhibitory activity. AB - Rapamycin, a complex macrolide and potent fungicide, immunosuppressant and anticancer agent, is a highly specific inhibitor of mammalian target of rapamycin (mTOR). Rapamycin has been shown to induce G1-phase cell cycle arrest in diverse tumor cell types, and its derivatives RAD001 and CCI-779 are currently in phase I and phase II clinical trials, respectively, as anticancer agents. In this study, we show that rapamycin induced the apoptotic death of JN-DSRCT-1 cells, the only available in vitro model for Desmoplastic Small Round Cell Tumors (DSRCT), while having only minor effects on their cell cycle. Rapamycin induced apoptosis by increasing the Bax : Bcl-xL ratio as a consequence of the concomitant downregulation of Bcl-xL and upregulation of Bax, both at the post transcriptional level. Rapamycin also downregulated the levels of EWS/WT1, the fusion protein characteristic of DSRCT. Transient transfection studies using kinase-dead and rapamycin-resistant forms of mTOR demonstrated that only the downregulation of Bcl-xL was caused by the mTOR inhibitory action of rapamycin, which prevented cap-dependent translation initiation, whereas Bax upregulation was induced by rapamycin through a mechanism independent of its mTOR inhibitory activity. Moreover, rapamycin treatment downregulated the mRNA and protein levels of the 26S p44.5 proteasome subunit, suggesting the involvement of the proteasome complex in the mechanisms of rapamycin-induced apoptosis. Treatment of JN-DSRCT-1 cells with MG-132, a proteasome specific inhibitor, also resulted in the induction of apoptosis through a similar increase in the Bax : Bcl-xL ratio specifically caused by inhibiting Bax degradation and turnover. These results suggested that rapamycin induces apoptosis by preventing the degradation of the Bax protein by the proteasome, and that this process is independent of mTOR inhibition. Furthermore, these results strongly support the introduction of the use of rapamycin as a cytotoxic agent for the treatment of DSRCT. PMID- 15782133 TI - AlphaVbeta3 integrin regulates heregulin (HRG)-induced cell proliferation and survival in breast cancer. AB - alpha(v)beta(3) integrin-overexpression in tumor associated vasculature is a marker of poor prognosis in breast cancer. A positive correlation between alpha(v)beta(3) integrin and overexpression of Heregulin (HRG), a growth factor associated with breast cancer aggressiveness was recently demonstrated. Here, we addressed the role of alpha(v)beta(3) in the proliferation and survival of HRG overexpressing breast cancer models. Expression of the RGD-binding integrins alpha(v)beta(3), alpha(v)beta(5) and alpha(v)beta(6) was assessed in the HRG overexpressing breast cancer cells MDA-MB-231, Hs578T (231/WT and Hs578T/WT, respectively) and derived cells transfected with the antisense orientation of the HRG-beta2 full-length cDNA (231/ASPOOL, 231/AS31 and Hs578T/AS15). Interestingly, only alpha(v)beta(3) expression was noticeably decreased by blockade of HRG expression in the 231/ASPOOL, 231/AS31 and Hs578T/AS15 cells. Small RGD-based peptidomimetic alpha(v)beta(3) antagonists significantly decreased cell viability and anchorage-dependent cell growth of HRG-overexpressing cells, while the low HRG-expressing 231/AS31 cells did not show a significant response. Mechanistically, functional blockade of alpha(v)beta(3) impaired HRG-promoted hyperactivation of ERK1/ERK2 MAPK without altering the activation of AKT. Flow cytometric analysis of the cell cycle demonstrated that alpha(v)beta(3) antagonists significantly decreased S- and G2/M-phase subpopulations of 231/WT and control 231/VEC cells. Comparable, this effect was linked to an increase in the levels and nuclear translocation of the CDKs inhibitor p27(Kip1). Besides downregulating alpha(v)beta(3) and its driven signaling, HRG blockade led to decreased levels of CYR61 in 231/ASPOOL and 231/AS31 cells. Considering that CYR61 is sufficient to upregulate the expression of alpha(v)beta(3), we then assessed alpha(v)beta(3) levels in MDA-MB-231 cell derivatives expressing the antisense orientation of the CYR61 cDNA (231/CYR61AS-5 and 231/CYR61AS-8). Remarkably, downregulation of CYR61 drastically decreased the levels of alpha(v)beta(3) in the 231/CYR61-5 and 231/CYR61-8 cells, providing further evidence of a key role for CYR61 in HRG-dependent alpha(v)beta(3) overexpression. Moreover, blockade of CYR61 expression impaired the HRG-induced hyperactivation of ERK1/ERK2 MAPK without altering the activation status of AKT in MDA-MB-231 cells, thus resembling the effects exerted by the downregulation of HRG expression as well as by functional blockade of alpha(v)beta(3). These results indicate that HRG is regulating alpha(v)beta(3) levels as well as alpha(v)beta(3) triggered signaling through its downstream effector, CYR61, in highly invasive breast cancer cells. Altogether, the data presented here provide evidence of a CYR61-regulatory role on alpha(v)beta(3) integrin expression in the modulation of uncontrolled growth of HRG-overexpressing breast carcinomas. This work supports additional studies concerning the use of integrin antagonists as dual therapeutic agents in breast cancer, targeting both, endothelial and tumor cells. PMID- 15782134 TI - Distinct roles for p53 transactivation and repression in preventing UCN-01 mediated abrogation of DNA damage-induced arrest at S and G2 cell cycle checkpoints. AB - The topoisomerase I inhibitor SN38 arrests cell cycle progression primarily in S or G(2) phases of the cell cycle in a p53-independent manner. The Chk1 inhibitor, 7-hydroxystaurosporine (UCN-01), overcomes both S and G(2) arrest preferentially in cells mutated for p53, driving cells through a lethal mitosis and thereby enhancing cytotoxicity. The mechanism by which p53 maintains S and G(2) arrest was investigated here. The p53 wild-type MCF10A cells were arrested in S phase by incubation with SN38 for 24 h. Subsequent incubation with UCN-01 failed to abrogate arrest. To examine the impact of p53, MCF10A cells were developed, which express the tetramerization domain of p53 to inhibit endogenous p53 function. These cells were attenuated in SN38-mediated induction of p21(WAF1), and UCN-01 induced S, but not G(2) progression. In contrast, MCF10A cells expressing short hairpin RNA to ablate p53 expression underwent both S and G(2) phase progression with UCN-01. The difference in G(2) progression was attributed to p53-mediated gene repression; the MCF10A cells expressing the tetramerization domain retained p53 protein and repressed both cyclin B and Chk1, while cells ablated for p53 did not repress these proteins. Hence, inhibition of p53 activator function permits S phase abrogation, while additional inhibition of p53 repressor function is required for abrogation of G(2) arrest. These studies provide a mechanistic explanation for how this therapeutic strategy can selectively target tumor cells. PMID- 15782135 TI - Caspase-8 sumoylation is associated with nuclear localization. AB - The cysteine protease caspase-8 plays a pivotal role in the initiation of different apoptotic pathways and controls the maturation and differentiation of various cell types including neurons, fibroblasts and lymphocytes. Specific substrates of caspase-8 are present in both the cytoplasm and the nucleus, which may determine the ultimate biological effect of caspase-8. However, the mechanisms regulating the cellular localization of caspase-8 are still unknown. We show here that, in contrast to other caspases such as caspase-9 and -3, caspase-8 can be sumoylated at lysine 156. This sumoylation (i) is associated with the nuclear localization of caspase-8 and (ii) did not impair caspase-8 activation. PMID- 15782136 TI - Genetic interactions between RAD51 and its paralogues for centrosome fragmentation and ploidy control, independently of the sensitivity to genotoxic stresses. AB - We evaluate here whether RAD51 and its paralogues XRCC2 and XRCC3 act via a common pathway for sensitivity to genotoxic stress, centrosome fragmentation and chromosome stability. We expressed the RAD51 dominant-negative SMRAD51 in irs1 and irs1SF cells, defective for XRCC2 and XRCC3, respectively, and in their corresponding wild-type cells (V79 and AA8, respectively). V79-SMRAD51 cells are sensitive to mitomycin C (MMC), but SMRAD51 did not further sensitize irs1 cells to MMC, showing that SMRAD51 and XRCC2 act on the same pathway for resistance to MMC. However, in contrast to irs1 and irs1SF cells, SMRAD51-V79 and SMRAD51-AA8 cells are not sensitive to gamma-rays or UV-C. Despite these differences in sensitivity, SMRAD51-expressing cells and xrcc2- or xrcc3-defective cells show similar increased levels of centrosome fragmentation. This spontaneous centrosome fragmentation is resistant to caffeine, suggesting that ATM and ATR are not involved. Consistent with centrosome fragmentation, increased aneuploidy was measured in irs1 and SMRAD51-expressing cells. Expression of SMRAD51 in irs1 or irs1SF cells did not increase further the frequency of multipolar cells. Thus, RAD51, XRCC2 and XRCC3 act in the same pathway for centrosome fragmentation, independently of the sensitivity to exogenous genotoxic stresses and of the ATM/ATR pathway. PMID- 15782137 TI - RKIP downregulates B-Raf kinase activity in melanoma cancer cells. AB - The Raf-MEK-ERK protein kinase cascade is a highly conserved signaling pathway that is pivotal in relaying environmental cues from the cell surface to the nucleus. Three Raf isoforms, which share great sequence and structure similarities, have been identified in mammalian cells. We have previously identified Raf kinase inhibitor protein (RKIP) as a negative regulator of the Raf MEK-ERK signaling pathway by specifically binding to the Raf-1 isoform. We show here that RKIP also antagonizes kinase activity of the B-Raf isoform. Yeast two hybrid and coimmunoprecipitation experiments indicated that RKIP specifically interacted with B-Raf. Ectopic expression of RKIP antagonized the kinase activity of B-Raf. We showed that the effects of RKIP on B-Raf functions were independent of its known inhibitory action on Raf-1. The expression levels of RKIP in melanoma cancer cell lines are low relative to primary melanocytes. Forced expression of RKIP partially reverted the oncogenic B-Raf kinase-transformed melanoma cancer cell line SK-Mel-28. The low expression of RKIP and its antagonistic action on B-Raf suggests that RKIP may play an important role in melanoma turmorgenesis. PMID- 15782138 TI - Differential roles for the coactivators CBP and p300 on TCF/beta-catenin-mediated survivin gene expression. AB - The inhibitor of apoptosis (IAP) protein survivin is highly expressed in cancers, but not in normal differentiated tissues. TCF/beta-catenin signaling has been reported to participate in the regulation of survivin transcription in colon cancer. We have recently characterized ICG-001, a small molecule specific inhibitor of the beta-catenin/Creb-binding protein (CBP) interaction. Inhibition of the beta-catenin/CBP interaction represses a subset of TCF/beta-catenin mediated transcription. ICG-001 potently inhibits survivin gene transcription and expression. ICG-001-mediated downregulation of survivin expression enhanced caspase-3 activity and apoptosis, which was rescued by overexpression of wild type but not mutant (C84A) survivin. Small interfering RNA and genetic reduction of CBP also decreased survivin expression. Chromatin immunoprecipitation assay confirmed that CBP is the crucial coactivator for TCF/beta-catenin-mediated survivin transcription. Furthermore, ICG-001-induced recruitment of p300 to the survivin promoter led to concomitant recruitment of SUMO-1, HDAC6 and PML proteins, which have been associated with transcriptional repression. These findings demonstrate that CBP and p300 play very distinct roles in survivin gene transcription. PMID- 15782139 TI - Connexin32 as a tumor suppressor gene in a metastatic renal cell carcinoma cell line. AB - Connexin genes expressing gap junction proteins have tumor-suppressive effects on primary cancers with certain cell specificity, but the suppressive effects on metastatic cancers are still conflicting. In this study, we show that connexin32 (Cx32) has a strong tumor-suppressive effect on a human metastatic renal cell carcinoma cell line (Caki-1 cell). Cx32 expression in Caki-1 cells reduced in vitro malignant phenotypes of the cells such as anchorage independency and invasion capacity. Furthermore, the Cx32 expression drastically reduced the development of Caki-1 cells in nude mice. We also determined that Cx32 reduced the malignant phenotypes in Caki-1 cells mainly through the inactivation of Src signaling. Especially, Cx32-dependent inactivation of Src decreased the production of vascular epithelial growth factor (VEGF) via the suppression of signal transducers and activators of transcription 3 (Stat3) activation, and we confirmed this result using short interfering RNA. In nude mice, Cx32-transfected Caki-1 cells showed lower serum level of VEGF comparing mock transfectant, and the development of the cells in nude mice positively related to the VEGF level. These data suggest that Cx32 acts as a tumor suppressor gene in Caki-1 cells and that the tumor-suppressive effect partly depends on the inhibition of Src-Stat3 VEGF signal pathway. PMID- 15782140 TI - Inhibition of ILK in PTEN-mutant human glioblastomas inhibits PKB/Akt activation, induces apoptosis, and delays tumor growth. AB - The tumor suppressor gene phosphatase and tensin homologue (PTEN) regulates the phosphatidylinositol-3'-kinase (PI3K) signaling pathway and has been shown to correlate with poor prognosis in high-grade astrocytomas when mutational inactivation or loss of the PTEN gene occurs. PTEN mutation leads to constitutive activation of protein kinase B (PKB)/Akt with phosphorylation at the PKB/Akt sites Thr-308 and Ser-473. Integrin-linked kinase (ILK) has been shown to regulate PKB/Akt activity with the loss of PTEN in prostate cancer. We now demonstrate that ILK activity regulates PKB/Akt activity in glioblastoma cells. The activity of ILK is constitutively elevated in a serum-independent manner in PTEN mutant cells, and transfection of wild-type PTEN under the control of an inducible promoter into mutant PTEN cells inhibits ILK activity. Transfection of ILK antisense (ILKAS) or exposure to a small-molecule ILK inhibitor suppresses the constitutive phosphorylation of PKB/Akt on Ser-473 in PTEN-mutant glioblastoma cell lines. In addition, the delivery of ILKAS to PTEN-negative glioblastoma cells resulted in apoptosis. Rag-2M mice bearing established ( approximately 100 mg) human U87MG glioblastoma tumors, treated QD x 5 for 3 consecutive weeks with ILKAS (i.p. 5 mg/kg), exhibited stable disease with < or =7% increase in tumor volume over the 3-week course of treatment. In contrast, animals treated with an oligonucleotide control or saline exhibited a >100% increase in tumor volume over the same time period. Our initial results indicate that therapeutic strategies targeting ILK may be beneficial in the treatment of glioblastomas. PMID- 15782141 TI - Induction of apoptosis by the synthetic retinoid MX3350-1 through extrinsic and intrinsic pathways in head and neck squamous carcinoma cells. AB - Retinoids have shown promise in cancer prevention and therapy. As some retinoids exhibit undesirable side effects, new retinoid analogs have been synthesized. In this study, we examined the effects of the retinoid MX3350-1 on human head and neck squamous cell carcinoma (HNSCC) cell lines. MX3350-1 suppressed the growth of 7/8 HNSCC cell lines by >65%. This inhibition appeared to be due to induction of apoptosis as revealed by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. Studies with cell line UMSCC17B indicated that apoptosis was induced within 1-2 days and involved activation of caspase-8, -9, and -3. Inhibitors of these caspases suppressed MX3350-1-induced apoptosis. MX3350-1 decreased the levels of antiapoptotic Bcl-2 and Bcl-XL, increased proapoptotic Bax, induced mitochondrial membrane permeabilization (MMP), and cytochrome c release from mitochondria to cytosol. The antioxidant butylated hydroxyanisol and the MMP inhibitor cyclosporin A (Cs A) blocked apoptosis induced by MX3350-1. In contrast, retinoid receptor antagonists failed to inhibit apoptosis. MX3350-1 increased the levels of Fas-ligand, Fas, and Fas-associated death domain, and enhanced activation of procaspase-8 and cleavage of its substrate Bid. Soluble Fas rescued the cells from MX3350-1-induced apoptosis. These results demonstrate that MX3350-1 induces apoptosis by activating both extrinsic and intrinsic apoptosis pathways and suggest that further studies on the potential of this retinoid for prevention and therapy of HNSCCs are warranted. PMID- 15782142 TI - Differential expression of the F-box proteins Skp2 and Skp2B in breast cancer. AB - Skp2 is an F-box protein involved in the ubiquitination and subsequent degradation of the cyclin-dependent kinase (Cdk) inhibitor p27. Skp2 has been reported to be overexpressed in a variety of cancer types and to correlate with poor prognosis. We have identified a novel isoform of Skp2 we named Skp2B, which differs from Skp2 only in the C-terminal domain and unlike Skp2 localizes to the cytoplasm. Here, we describe the relative expression of both Skp2 and Skp2B in breast cancer cell lines and in primary breast cancers using quantitative real time RT-PCR. We show that Skp2B mRNA is expressed 10-fold less than Skp2 mRNA in the immortalized but non-transformed breast cell line, 184B5. However, Skp2B is overexpressed as frequently as Skp2, and to higher levels than Skp2 in breast cancer cell lines and primary cancers. Further, we show that cytoplasmic staining is frequent in primary breast cancers. In addition, we found that xenografts expressing Skp2B grow faster than xenografts expressing low levels of Skp2B, and that this effect is independent of p27 degradation. These findings therefore suggest that Skp2B overexpression is also observed in breast cancers and identify Skp2B as a putative oncogene. PMID- 15782143 TI - Vasculostatin, a proteolytic fragment of brain angiogenesis inhibitor 1, is an antiangiogenic and antitumorigenic factor. AB - Brain angiogenesis inhibitor 1 (BAI1) is a transmembrane protein with unknown function expressed primarily in normal but not tumoral brain. The finding of thrombospondin type 1 repeats in its extracellular domain suggested an antiangiogenic function, but the mechanisms by which a transmembrane receptor could inhibit angiogenesis remained unexplained. Here we demonstrate that BAI1 is proteolytically cleaved at a conserved G-protein-coupled receptor proteolytic cleavage site (GPS), releasing its 120 kDa extracellular domain. We named this secreted fragment Vasculostatin as it inhibited migration of endothelial cells in vitro and dramatically reduced in vivo angiogenesis. Both constitutive and doxycycline-induced expression of Vasculostatin elicited dose-dependent suppression of tumor growth and vascular density in mice, implicating Vasculostatin in the regulation of vascular homeostasis and tumor prevention. Generation of a soluble antiangiogenic factor by cleavage of a pre-existing transmembrane protein represents a novel mechanism for regulating vascular homeostasis and preventing tumorigenesis. Modulation of this cleavage or delivery of Vasculostatin may constitute novel treatment modalities for cancer and other diseases of aberrant angiogenesis, especially in the brain. PMID- 15782144 TI - PTPL1 is a direct transcriptional target of EWS-FLI1 and modulates Ewing's Sarcoma tumorigenesis. AB - Ewing's Sarcoma family tumors (ESFT) are characterized by a translocation t(11:22) forming an aberrant transcription factor EWS-FLI1. Protein tyrosine phosphatase L1 (PTPL1) was identified as a gene upregulated by EWS-FLI1 in transfected cells by microarray. Our results show that PTPL1 is a transcriptional target of EWS-FLI1 both by chromatin immunoprecipitation and promoter activation studies. We demonstrate that PTPL1 is highly expressed in ESFT cells and patient tumors compared with normal tissues, with a trend towards higher expression in metastatic versus primary tumors. Reduction of PTPL1 protein in ESFT cells correlated with a significant reduction in both monolayer and soft-agar cell growth. In addition, these PTPL1-reduced cells were more sensitive to etoposide induced apoptosis than the controls. We therefore report a novel transcriptional activation of a phosphatase involved in the oncogenesis of ESFT. Increasing interest in specific phosphatase inhibitors would allow PTPL1 to be evaluated as a therapeutic target in ESFT. PMID- 15782146 TI - Novel at the library. PMID- 15782145 TI - SSBP2, a candidate tumor suppressor gene, induces growth arrest and differentiation of myeloid leukemia cells. AB - Acute myelogenous leukemia (AML) is the most common leukemia in adults with clonal proliferation of myeloid stem cells. Two or more cooperating mechanisms, namely block in differentiation, enhanced proliferation and resistance to programmed cell death, underlie this neoplastic transformation. Nonrandom, complete and partial deletions of chromosome 5 are common anomalies in AML. Using positional cloning strategies, we characterized an evolutionarily conserved candidate myeloid leukemia suppressor gene encoding sequence-specific single stranded DNA binding protein 2 (SSBP2) from chromosome 5q13.3, a locus that is frequently deleted in AML. Recent studies in Drosophila and Xenopus demonstrate a pivotal role for SSBPs in embryonic differentiation. In mammals, SSBP2 is one of three highly related and ubiquitously expressed genes. Here, we identify frequent loss of SSBP2 protein expression in human AML cell lines using highly specific antibodies. Furthermore, inducible expression of SSBP2 in the AML cell line U937 leads to loss of clonogenicity, G1 arrest and partial differentiation. Remarkably, inducible expression of SSBP2 is accompanied by downregulation of C MYC expression. Our findings are consistent with human SSBP2 being a novel regulator of hematopoietic growth and differentiation, whose loss confers a block in differentiation advantage to myeloid leukemic cells. PMID- 15782147 TI - Centromeric heterochromatin comes clean with DNA methylation. PMID- 15782148 TI - Shotgun mass spec goes independent. PMID- 15782149 TI - A rapid method for determining protein kinase phosphorylation specificity. AB - Selection of target substrates by protein kinases is strongly influenced by the amino acid sequence surrounding the phosphoacceptor site. Identification of the preferred peptide phosphorylation motif for a given kinase permits the production of efficient peptide substrates and greatly simplifies the mapping of phosphorylation sites in protein substrates. Here we describe a combinatorial peptide library method that allows rapid generation of phosphorylation motifs for serine/threonine kinases. PMID- 15782150 TI - Sulforhodamine 101 as a specific marker of astroglia in the neocortex in vivo. AB - Glial cells have been identified as key signaling components in the brain; however, methods to investigate their structure and function in vivo have been lacking. Here, we describe a new, highly selective approach for labeling astrocytes in intact rodent neocortex that allows in vivo imaging using two photon microscopy. The red fluorescent dye sulforhodamine 101 (SR101) was specifically taken up by protoplasmic astrocytes after brief exposure to the brain surface. Specificity was confirmed by immunohistochemistry. In addition, SR101 labeled enhanced green fluorescent protein (EGFP)-expressing astrocytes but not microglial cells in transgenic mice. We used SR101 labeling to quantify morphological characteristics of astrocytes and to visualize their close association with the cortical microvasculature. Furthermore, by combining this method with calcium indicator loading of cell populations, we demonstrated distinct calcium dynamics in astroglial and neuronal networks. We expect SR101 staining to become a principal tool for investigating astroglia in vivo. PMID- 15782152 TI - A custom microarray platform for analysis of microRNA gene expression. AB - MicroRNAs are short, noncoding RNA transcripts that post-transcriptionally regulate gene expression. Several hundred microRNA genes have been identified in Caenorhabditis elegans, Drosophila, plants and mammals. MicroRNAs have been linked to developmental processes in C. elegans, plants and humans and to cell growth and apoptosis in Drosophila. A major impediment in the study of microRNA function is the lack of quantitative expression profiling methods. To close this technological gap, we have designed dual-channel microarrays that monitor expression levels of 124 mammalian microRNAs. Using these tools, we observed distinct patterns of expression among adult mouse tissues and embryonic stem cells. Expression profiles of staged embryos demonstrate temporal regulation of a large class of microRNAs, including members of the let-7 family. This microarray technology enables comprehensive investigation of microRNA expression, and furthers our understanding of this class of recently discovered noncoding RNAs. PMID- 15782151 TI - Automated approach for quantitative analysis of complex peptide mixtures from tandem mass spectra. AB - To take advantage of the potential quantitative benefits offered by tandem mass spectrometry, we have modified the method in which tandem mass spectrum data are acquired in 'shotgun' proteomic analyses. The proposed method is not data dependent and is based on the sequential isolation and fragmentation of precursor windows (of 10 m/z) within the ion trap until a desired mass range has been covered. We compared the quantitative figures of merit for this method to those for existing strategies by performing an analysis of the soluble fraction of whole-cell lysates from yeast metabolically labeled in vivo with (15)N. To automate this analysis, we modified software (RelEx) previously written in the Yates lab to generate chromatograms directly from tandem mass spectra. These chromatograms showed improvements in signal-to-noise ratio of approximately three to fivefold over corresponding chromatograms generated from mass spectrometry scans. In addition, to demonstrate the utility of the data-independent acquisition strategy coupled with chromatogram reconstruction from tandem mass spectra, we measured protein expression levels in two developmental stages of Caenorhabditis elegans. PMID- 15782154 TI - Exploiting the 21st amino acid-purifying and labeling proteins by selenolate targeting. AB - Selenium is essential to human life and occurs in selenoproteins as selenocysteine (Sec), the 21st amino acid. The selenium atom endows selenocysteine with unique biochemical properties, including a low pK(a) and a high reactivity with many electrophilic agents. Here we describe the introduction of selenocysteine into recombinant non-selenoproteins produced in Escherichia coli, as part of a small tetrapeptide motif at the C terminus. This selenocysteine-containing motif could subsequently be used as a protein tag for purification of the recombinant protein, selenolate-targeted labeling with fluorescent compounds or radiolabeling with either gamma-emitting (75)Se or short lived positron emitters such as (11)C. The results presented here thus show how a wide range of biotechnological applications can be developed starting from the insertion of selenocysteine into proteins. PMID- 15782153 TI - In vitro selection and prediction of TIP47 protein-interaction interfaces. AB - We present a new method for the rapid identification of amino acid residues that contribute to protein-protein interfaces. Tail-interacting protein of 47 kDa (TIP47) binds Rab9 GTPase and the cytoplasmic domains of mannose 6-phosphate receptors and is required for their transport from endosomes to the Golgi apparatus. Cysteine mutations were incorporated randomly into TIP47 by expression in Escherichia coli cells harboring specific misincorporator tRNAs. We made use of the ability of the native TIP47 protein to protect 48 cysteine probes from chemical modification by iodoacetamide as a means to obtain a surface map of TIP47, revealing the identity of surface-localized, hydrophobic residues that are likely to participate in protein-protein interactions. Direct mutation of predicted interface residues confirmed that the protein had altered binding affinity for the mannose 6-phosphate receptor. TIP47 mutants with enhanced or diminished affinities were also selected by affinity chromatography. These methods were validated in comparison with the protein's crystal structure, and provide a powerful means to predict protein-protein interaction interfaces. PMID- 15782155 TI - Whole-genome fractionation rapidly purifies DNA from centromeric regions. AB - The condensed centromeric regions of higher eukaryotic chromosomes contain satellite sequences, transposons and retroelements, as well as transcribed genes that perform a variety of functions. These chromosomal domains nucleate kinetochores, mediate sister chromatid cohesion and inhibit recombination, yet their characterization has often lagged behind that of chromosome arms. Here, we describe a whole-genome fractionation technique that rapidly identifies bacterial artificial chromosome (BAC) clones derived from plant centromeric regions. This approach, which relies on hybridization of methylated genomic DNA, revealed BACs that correspond to the genetically mapped and sequenced Arabidopsis thaliana centromeric regions. Extending this method to other species in the Brassicaceae family identified centromere-linked clones and provided genome-wide estimates of methylated DNA abundance. Sequencing these clones will elucidate the changes that occur during plant centromere evolution. This genomic fractionation technique could identify centromeric DNA in genomes with similar methylation and repetitive DNA content, including those from crops and mammals. PMID- 15782158 TI - Using protein-DNA chimeras to detect and count small numbers of molecules. AB - We describe general methods to detect and quantify small numbers of specific molecules. We redirected self-splicing protein inteins to create 'tadpoles', chimeric molecules comprised of a protein head covalently coupled to an oligonucleotide tail. We made different classes of tadpoles that bind specific targets, including Bacillus anthracis protective antigen and the enzyme cofactor biotin. We measured the amount of bound target by quantifying DNA tails by T7 RNA polymerase runoff transcription and real-time polymerase chain reaction (PCR) evaluated by rigorous statistical methods. These assays had a dynamic range of detection of more than 11 orders of magnitude and distinguished numbers of molecules that differed by as little as 10%. At their low limit, these assays were used to detect as few as 6,400 protective antigen molecules, 600 biotin molecules and 150 biotinylated protein molecules. In crudely fractionated human serum, the assays were used to detect as few as 32,000 protective antigen molecules. Tadpoles thus enable sensitive detection and precise quantification of molecules other than DNA and RNA. PMID- 15782159 TI - Polyene-lipids: a new tool to image lipids. AB - Microscopy of lipids in living cells is currently hampered by a lack of adequate fluorescent tags. The most frequently used tags, NBD and BODIPY, strongly influence the properties of lipids, yielding analogs with quite different characteristics. Here, we introduce polyene-lipids containing five conjugated double bonds as a new type of lipid tag. Polyene-lipids exhibit a unique structural similarity to natural lipids, which results in minimal effects on the lipid properties. Analyzing membrane phase partitioning, an important biophysical and biological property of lipids, we demonstrated the superiority of polyene lipids to both NBD- and BODIPY-tagged lipids. Cells readily take up various polyene-lipid precursors and generate the expected end products with no apparent disturbance by the tag. Applying two-photon excitation microscopy, we imaged the distribution of polyene-lipids in living mammalian cells. For the first time, ether lipids, important for the function of the brain, were successfully visualized. PMID- 15782160 TI - Mapping DNA-protein interactions in large genomes by sequence tag analysis of genomic enrichment. AB - Identifying the chromosomal targets of transcription factors is important for reconstructing the transcriptional regulatory networks underlying global gene expression programs. We have developed an unbiased genomic method called sequence tag analysis of genomic enrichment (STAGE) to identify the direct binding targets of transcription factors in vivo. STAGE is based on high-throughput sequencing of concatemerized tags derived from target DNA enriched by chromatin immunoprecipitation. We first used STAGE in yeast to confirm that RNA polymerase III genes are the most prominent targets of the TATA-box binding protein. We optimized the STAGE protocol and developed analysis methods to allow the identification of transcription factor targets in human cells. We used STAGE to identify several previously unknown binding targets of human transcription factor E2F4 that we independently validated by promoter-specific PCR and microarray hybridization. STAGE provides a means of identifying the chromosomal targets of DNA-associated proteins in any sequenced genome. PMID- 15782161 TI - LAMP, a new imaging assay of gap junctional communication unveils that Ca2+ influx inhibits cell coupling. AB - Using a new class of photo-activatible fluorophores, we have developed a new imaging technique for measuring molecular transfer rates across gap junction connexin channels in intact living cells. This technique, named LAMP, involves local activation of a molecular fluorescent probe, NPE-HCCC2/AM, to optically label a cell. Subsequent dye transfer through gap junctions from labeled to unlabeled cells was quantified by fluorescence microscopy. Additional uncagings after prior dye transfers reached equilibrium enabled multiple measurements of dye transfer rates in the same coupled cell pair. Measurements in the same cell pair minimized variation due to differences in cell volume and number of gap junctions, allowing us to track acute changes in gap junction permeability. We applied the technique to study the regulation of gap junction coupling by intracellular Ca(2+) ([Ca(2+)](i)). Although agonist or ionomycin exposure can raise bulk [Ca(2+)](i) to levels higher than those caused by capacitative Ca(2+) influx, the LAMP assay revealed that only Ca(2+) influx through the plasma membrane store-operated Ca(2+) channels strongly reduced gap junction coupling. The noninvasive and quantitative nature of this imaging technique should facilitate future investigations of the dynamic regulation of gap junction communication. PMID- 15782162 TI - The matrix refolded. PMID- 15782163 TI - Tadpoles by the tail. PMID- 15782164 TI - Illuminating gap junctions. PMID- 15782165 TI - Brilliant lipids. PMID- 15782166 TI - The invertor knock-in conditional chromosomal translocation mimic. AB - Knock-in models of tumor-specific chromosomal translocations can generate lethal mutations. To circumvent this, a new conditional gene fusion model has been developed (invertor mice) and exemplified with the Ews-ERG fusion oncogene. An ERG segment, flanked by loxP sites, was knocked in to an intron of the Ews gene but in an inverted transcription orientation and lineage-specific Ews-ERG fusion created by Cre-mediated inversion. This invertor method is a completely conditional approach, applicable to any gene fusion, to emulate effects of translocations found in human cancers. PMID- 15782169 TI - 5,000 RNAi experiments on a chip. PMID- 15782170 TI - SNPs made routine. PMID- 15782171 TI - The age of high-throughput microRNA profiling. PMID- 15782172 TI - Genotyping over 100,000 SNPs on a pair of oligonucleotide arrays. AB - We present a genotyping method for simultaneously scoring 116,204 SNPs using oligonucleotide arrays. At call rates >99%, reproducibility is >99.97% and accuracy, as measured by inheritance in trios and concordance with the HapMap Project, is >99.7%. Average intermarker distance is 23.6 kb, and 92% of the genome is within 100 kb of a SNP marker. Average heterozygosity is 0.30, with 105,511 SNPs having minor allele frequencies >5%. PMID- 15782173 TI - A highly informative SNP linkage panel for human genetic studies. AB - We have developed a highly informative set of single-nucleotide polymorphism (SNP) assays designed for linkage mapping of the human genome. These assays were developed on a robust multiplexed assay system to provide a combination of very high accuracy and data completeness with high throughput for linkage studies. The linkage panel is comprised of approximately 4,700 SNPs with 0.39 average minor allele frequency and 624-kb average spacing. Based on almost 2 million genotypes, data quality was shown to be extremely high, with a 99.94% call rate, >99.99% reproducibility and 99.995% genotypes consistent with mendelian inheritance. We constructed a genetic map with an average 1.5-cM resolution using series of 28 CEPH pedigrees. The relative information content of this panel was higher than those of commonly used STR marker panels. The potent combination of this SNP linkage panel with the multiplexed assay system provides a previously unattainable level of performance for linkage studies. PMID- 15782174 TI - Identifying and quantifying in vivo methylation sites by heavy methyl SILAC. AB - Protein methylation is a stable post-translational modification (PTM) with important biological functions. It occurs predominantly on arginine and lysine residues with varying numbers of methyl groups, such as mono-, di- or trimethyl lysine. Existing methods for identifying methylation sites are laborious, require large amounts of sample and cannot be applied to complex mixtures. We have previously described stable isotope labeling by amino acids in cell culture (SILAC) for quantitative comparison of proteomes. In heavy methyl SILAC, cells metabolically convert [(13)CD(3)]methionine to the sole biological methyl donor, [(13)CD(3)]S-adenosyl methionine. Heavy methyl groups are fully incorporated into in vivo methylation sites, directly labeling the PTM. This provides markedly increased confidence in identification and relative quantitation of protein methylation by mass spectrometry. Using antibodies targeted to methylated residues and analysis by liquid chromatography-tandem mass spectrometry, we identified 59 methylation sites, including previously unknown sites, considerably extending the number of in vivo methylation sites described in the literature. PMID- 15782175 TI - RNAi living-cell microarrays for loss-of-function screens in Drosophila melanogaster cells. AB - RNA interference (RNAi)-mediated loss-of-function screening in Drosophila melanogaster tissue culture cells is a powerful method for identifying the genes underlying cell biological functions and for annotating the fly genome. Here we describe the development of living-cell microarrays for screening large collections of RNAi-inducing double-stranded RNAs (dsRNAs) in Drosophila cells. The features of the microarrays consist of clusters of cells 200 mum in diameter, each with an RNAi-mediated depletion of a specific gene product. Because of the small size of the features, thousands of distinct dsRNAs can be screened on a single chip. The microarrays are suitable for quantitative and high-content cellular phenotyping and, in combination screens, for the identification of genetic suppressors, enhancers and synthetic lethal interactions. We used a prototype cell microarray with 384 different dsRNAs to identify previously unknown genes that affect cell proliferation and morphology, and, in a combination screen, that regulate dAkt/dPKB phosphorylation in the absence of dPTEN expression. PMID- 15782176 TI - Simultaneous, coincident optical trapping and single-molecule fluorescence. AB - We constructed a microscope-based instrument capable of simultaneous, spatially coincident optical trapping and single-molecule fluorescence. The capabilities of this apparatus were demonstrated by studying the force-induced strand separation of a dye-labeled, 15-base-pair region of double-stranded DNA (dsDNA), with force applied either parallel ('unzipping' mode) or perpendicular ('shearing' mode) to the long axis of the region. Mechanical transitions corresponding to DNA hybrid rupture occurred simultaneously with discontinuous changes in the fluorescence emission. The rupture force was strongly dependent on the direction of applied force, indicating the existence of distinct unbinding pathways for the two force loading modes. From the rupture force histograms, we determined the distance to the thermodynamic transition state and the thermal off rates in the absence of load for both processes. PMID- 15782177 TI - LigAmp for sensitive detection of single-nucleotide differences. AB - We developed the LigAmp assay for sensitive detection and accurate quantification of viruses and cells with single-base mutations. In LigAmp, two oligonucleotides are hybridized adjacently to a DNA template. One oligonucleotide matches the target sequence and contains a probe sequence. If the target sequence is present, the oligonucleotides are ligated together and detected using real-time PCR. LigAmp detected KRAS2 mutant DNA at 0.01% in mixtures of different cell lines. KRAS2 mutations were also detected in pancreatic duct juice from patients with pancreatic cancer. LigAmp detected the K103N HIV-1 drug resistance mutation at 0.01% in plasmid mixtures and at approximately 0.1% in DNA amplified from plasma HIV-1. Detection in both systems is linear over a broad dynamic range. Preliminary evidence indicates that reactions can be multiplexed. This assay may find applications in the diagnosis of genetic disorders and the management of patients with cancer and infectious diseases. PMID- 15782178 TI - Cell-free protein production and labeling protocol for NMR-based structural proteomics. AB - Structural proteomics requires robust, scalable methods. Here we describe a wheat germ cell-free platform for protein production that supports efficient NMR structural studies of eukaryotic proteins and offers advantages over cell-based methods. To illustrate this platform, we describe its application to a specific target (At3g01050.1) from Arabidopsis thaliana. After cloning the target gene into a specialized plasmid, we carry out a small-scale (50 mul) in vitro sequential transcription and translation trial to ascertain the level of protein production and solubility. Next, we prepare mRNA for use in a 4-ml semicontinuous cell-free translation reaction to incorporate (15)N-labeled amino acids into a protein sample that we purify and test for suitability for NMR structural analysis. We then repeat the cell-free approach with (13)C,(15)N-labeled amino acids to prepare a doubly labeled sample. The three-dimensional (3D) structure of At3g01050.1 shows that this protein is an unusual member of the beta-grasp protein family. PMID- 15782179 TI - Microarray-based, high-throughput gene expression profiling of microRNAs. AB - MicroRNAs (miRNAs) are small regulatory RNAs that serve fundamental biological roles across eukaryotic species. We describe a new method for high-throughput miRNA detection. The technique is termed the RNA-primed, array-based Klenow enzyme (RAKE) assay, because it involves on-slide application of the Klenow fragment of DNA polymerase I to extend unmodified miRNAs hybridized to immobilized DNA probes. We used RAKE to study human cell lines and brain tumors. We show that the RAKE assay is sensitive and specific for miRNAs and is ideally suited for rapid expression profiling of all known miRNAs. RAKE offers unique advantages for specificity over northern blots or other microarray-based expression profiling platforms. Furthermore, we demonstrate that miRNAs can be isolated and profiled from formalin-fixed paraffin-embedded tissue, which opens up new opportunities for analyses of small RNAs from archival human tissue. The RAKE assay is theoretically versatile and may be used for other applications, such as viral gene profiling. PMID- 15782182 TI - New eyes to see texture in ligand efficacy. PMID- 15782183 TI - Stem cell culture, one step at a time. PMID- 15782184 TI - Stem cell proliferative history in tissue revealed by temporal halogenated thymidine analog discrimination. AB - Detection of proliferating cells based on bromodeoxyuridine (BrdU) incorporation and determination of phenotype by immunofluorescence labeling are standard approaches for studying stem and progenitor cell populations in developing and adult tissue as well as in histopathology studies. We describe incorporation of different halogenated thymidine analogs for temporal discrimination of cell cycle in the rat. With equimolar delivery, these analogs are suitable for quantitative histological studies including assessment of the regulation of proliferation, clonal analysis and simultaneous profiling of cell phenotype relative to proliferative history. PMID- 15782185 TI - A FlAsH-based FRET approach to determine G protein-coupled receptor activation in living cells. AB - Fluorescence resonance energy transfer (FRET) from cyan to yellow fluorescent proteins (CFP/YFP) is a well-established method to monitor protein-protein interactions or conformational changes of individual proteins. But protein functions can be perturbed by fusion of large tags such as CFP and YFP. Here we use G protein-coupled receptor (GPCR) activation in living cells as a model system to compare YFP with the small, membrane-permeant fluorescein derivative with two arsen-(III) substituents (fluorescein arsenical hairpin binder; FlAsH) targeted to a short tetracysteine sequence. Insertion of CFP and YFP into human adenosine A(2A) receptors allowed us to use FRET to monitor receptor activation but eliminated coupling to adenylyl cyclase. The CFP/FlAsH-tetracysteine system gave fivefold greater agonist-induced FRET signals, similar kinetics (time constant of 66-88 ms) and perfectly normal downstream signaling. Similar results were obtained for the mouse alpha(2A)-adrenergic receptor. Thus, FRET from CFP to FlAsH reports GPCR activation in living cells without disturbing receptor function and shows that the small size of the tetracysteine-biarsenical tag can be decisively advantageous. PMID- 15782186 TI - Real-time monitoring of receptor and G-protein interactions in living cells. AB - G protein-coupled receptors (GPCRs) represent the largest family of proteins involved in signal transduction. Here we present a bioluminescence resonance energy transfer (BRET) assay that directly monitors in real time the early interactions between human GPCRs and their cognate G-protein subunits in living human cells. In addition to detecting basal precoupling of the receptors to Galpha-, Gbeta- and Ggamma-subunits, BRET measured very rapid ligand-induced increases in the interaction between receptor and Galphabetagamma-complexes (t(1/2) approximately 300 ms) followed by a slower (several minutes) decrease, reflecting receptor desensitization. The agonist-promoted increase in GPCR Gbetagamma interaction was highly dependent on the identity of the Galpha-subunit present in the complex. Therefore, this G protein-activity biosensor provides a novel tool to directly probe the dynamics and selectivity of receptor-mediated, G protein activation-deactivation cycles that could be advantageously used to identify ligands for orphan GPCRs. PMID- 15782187 TI - Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells. AB - Human embryonic stem cells (hESCs) are routinely cultured on fibroblast feeder layers or in fibroblast-conditioned medium (CM). Bone morphogenetic proteins (BMPs) have previously been shown to induce hESC differentiation, in apparent contrast to mouse embryonic stem (ES) cells, in which BMP4 synergizes with leukemia inhibitory factor (LIF) to maintain self-renewal. Here we demonstrate that hESCs cultured in unconditioned medium (UM) are subjected to high levels of BMP signaling activity, which is reduced in CM. The BMP antagonist noggin synergizes with basic fibroblast growth factor (bFGF) to repress BMP signaling and sustain undifferentiated proliferation of hESCs in the absence of fibroblasts or CM. These findings suggest a basic difference in the self-renewal mechanism between mouse and human ES cells and simplify the culture of hESCs. PMID- 15782188 TI - A new method for C-terminal sequence analysis in the proteomic era. AB - The overall study of post-translational modifications (PTMs) of proteins is gaining strong interest. Beside phosphorylation and glycosylation, truncations of the nascent polypeptide chain at the amino or carboxy terminus are by far the most common types of PTMs in proteins. In contrast to the analysis of phosphorylation and glycosylation sites, relatively little attention has been paid to the development of approaches for the systematic analysis of proteolytic processing events. Here we present a new mass spectrometry (MS)-based strategy that allows the identification of the C-terminal sequence of proteins. The method can be directly applied to proteins cleaved with cyanogen bromide (CNBr) and purified either by SDS-PAGE, by two-dimensional (2D) PAGE or in solution, and it therefore eliminates the need for specific isolation of the C-terminal peptide. Using Shewanella oneidensis as a model system, we have demonstrated that this approach can be used for C-terminal sequence analysis at a proteomic scale. We also applied the method to study the C-terminal proteolytic processing of procardosin A. PMID- 15782189 TI - Protein photo-cross-linking in mammalian cells by site-specific incorporation of a photoreactive amino acid. AB - We report a method of photo-cross-linking proteins in mammalian cells, which is based on site-specific incorporation of a photoreactive amino acid, p-benzoyl-L phenylalanine (pBpa), through the use of an expanded genetic code. To analyze the cell signaling interactions involving the adaptor protein Grb2, pBpa was incorporated in its Src homology 2 (SH2) domain. The human GRB2 gene with an amber codon was introduced into Chinese hamster ovary (CHO) cells, together with the genes for the Bacillus stearothermophilus suppressor tRNA(Tyr) and a pBpa specific variant of Escherichia coli tyrosyl-tRNA synthetase (TyrRS). The Grb2 variant with pBpa in the amber position was synthesized when pBpa was included in the growth medium. Upon exposure of cells to 365-nm light, protein variants containing pBpa in the positions proximal to the ligand-binding pocket were cross linked with the transiently expressed epidermal growth factor (EGF) receptor in the presence of an EGF stimulus. Cross-linked complexes with endogenous proteins were also detected. In vivo photo-cross-linking with pBpa incorporated in proteins will be useful for studying protein-protein interactions in mammalian cells. PMID- 15782190 TI - Pulse proteolysis: a simple method for quantitative determination of protein stability and ligand binding. AB - Thermodynamic stability is fundamental to the biology of proteins. Information on protein stability is essential for studying protein structure and folding and can also be used indirectly to monitor protein-ligand or protein-protein interactions. While clearly valuable, the experimental determination of a protein's stability typically requires biophysical instrumentation and substantial quantities of purified protein, which has limited the use of this technique as a general laboratory method. We report here a simple new method for determining protein stability by using pulse proteolysis with varying concentrations of denaturant. Pulse proteolysis is designed to digest only the unfolded proteins in an equilibrium mixture of folded and unfolded proteins that relaxes on a time scale longer than the proteolytic pulse. We used this method to study the stabilities of Escherichia coli ribonuclease H and its variants, both in purified form and directly from cell lysates. The DeltaG(unf) degrees values obtained by this technique were in agreement with those determined by traditional methods. We also successfully used this method to monitor the binding of maltose binding protein to maltose, as well as to rapidly screen cognate ligands for this protein. The simplicity of pulse proteolysis suggests that it is an excellent strategy for the high-throughput determination of protein stability in protein engineering and drug discovery applications. PMID- 15782194 TI - Resonating to the music of ubiquitination. PMID- 15782195 TI - Real-time monitoring of ubiquitination in living cells by BRET. AB - Ubiquitin has emerged as an important regulator of protein stability and function in organisms ranging from yeast to mammals. The ability to detect in situ changes in protein ubiquitination without perturbing the physiological environment of cells would be a major step forward in understanding the ubiquitination process and its consequences. Here, we describe a new method to study this dynamic post translational modification in intact human embryonic kidney cells. Using bioluminescence resonance energy transfer (BRET), we measured the ubiquitination of beta-arrestin 2, a regulatory protein implicated in the modulation of G protein-coupled receptors. In addition to allowing the detection of basal and GPCR-regulated ubiquitination of beta-arrestin 2 in living cells, real-time BRET measurements permitted the recording of distinct ubiquitination kinetics that are dictated by the identity of the activated receptor. The ubiquitination BRET assay should prove to be a useful tool for studying the dynamic ubiquitination of proteins and for understanding which cellular functions are regulated by this post-translational event. PMID- 15782196 TI - Three-chromophore FRET microscopy to analyze multiprotein interactions in living cells. AB - Nearly every major process in a cell is carried out by assemblies of multiple dynamically interacting protein molecules. To study multi-protein interactions within such molecular machineries, we have developed a fluorescence microscopy method called three-chromophore fluorescence resonance energy transfer (3-FRET). This method allows analysis of three mutually dependent energy transfer processes between the fluorescent labels, such as cyan, yellow and monomeric red fluorescent proteins. Here, we describe both theoretical and experimental approaches that discriminate the parallel versus the sequential energy transfer processes in the 3-FRET system. These approaches were established in vitro and in cultured mammalian cells, using chimeric proteins consisting of two or three fluorescent proteins linked together. The 3-FRET microscopy was further applied to the analysis of three-protein interactions in the constitutive and activation dependent complexes in single endosomal compartments. These data highlight the potential of 3-FRET microscopy in studies of spatial and temporal regulation of signaling processes in living cells. PMID- 15782197 TI - High-throughput localization of functional elements by quantitative chromatin profiling. AB - Identification of functional, noncoding elements that regulate transcription in the context of complex genomes is a major goal of modern biology. Localization of functionality to specific sequences is a requirement for genetic and computational studies. Here, we describe a generic approach, quantitative chromatin profiling, that uses quantitative analysis of in vivo chromatin structure over entire gene loci to rapidly and precisely localize cis-regulatory sequences and other functional modalities encoded by DNase I hypersensitive sites. To demonstrate the accuracy of this approach, we analyzed approximately 300 kilobases of human genome sequence from diverse gene loci and cleanly delineated functional elements corresponding to a spectrum of classical cis regulatory activities including enhancers, promoters, locus control regions and insulators as well as novel elements. Systematic, high-throughput identification of functional elements coinciding with DNase I hypersensitive sites will substantially expand our knowledge of transcriptional regulation and should simplify the search for noncoding genetic variation with phenotypic consequences. PMID- 15782198 TI - In situ genotyping individual DNA molecules by target-primed rolling-circle amplification of padlock probes. AB - Methods are needed to study single molecules to reveal variability, interactions and mechanisms that may go undetected at the level of populations of molecules. We describe here an integrated series of reaction steps that allow individual nucleic acid molecules to be detected with excellent specificity. Oligonucleotide probes are circularized after hybridization to target sequences that have been prepared so that localized amplification reactions can be initiated from the target molecules. The process results in strong, discrete detection signals anchored to the target molecules. We use the method to observe the distribution, within and among human cells, of individual normal and mutant mitochondrial genomes that differ at a single nucleotide position. PMID- 15782200 TI - Production of complex nucleic acid libraries using highly parallel in situ oligonucleotide synthesis. AB - Generation of complex libraries of defined nucleic acid sequences can greatly aid the functional analysis of protein and gene function. Previously, such studies relied either on individually synthesized oligonucleotides or on cellular nucleic acids as the starting material. As each method has disadvantages, we have developed a rapid and cost-effective alternative for construction of small fragment DNA libraries of defined sequences. This approach uses in situ microarray DNA synthesis for generation of complex oligonucleotide populations. These populations can be recovered and either used directly or immortalized by cloning. From a single microarray, a library containing thousands of unique sequences can be generated. As an example of the potential applications of this technology, we have tested the approach for the production of plasmids encoding short hairpin RNAs (shRNAs) targeting numerous human and mouse genes. We achieved high-fidelity clone retrieval with a uniform representation of intended library sequences. PMID- 15782204 TI - Transcriptome PETs: a genome's best friends. PMID- 15782205 TI - Engineering of a vaccinia virus bacterial artificial chromosome in Escherichia coli by bacteriophage lambda-based recombination. AB - The large capacity of vaccinia virus (VAC) for added DNA, cytoplasmic expression and broad host range make it a popular choice for gene delivery, despite the burdensome need for multiple plaque purifications to isolate recombinants. Here we describe how a bacterial artificial chromosome (BAC) containing the entire VAC genome can be engineered in Escherichia coli by homologous recombination using bacteriophage lambda-encoded enzymes. The engineered VAC genomes can then be used to produce clonally pure recombinant viruses in mammalian cells without the need for plaque purification. PMID- 15782206 TI - Site-specific labeling of cell surface proteins with biophysical probes using biotin ligase. AB - We report a highly specific, robust and rapid new method for labeling cell surface proteins with biophysical probes. The method uses the Escherichia coli enzyme biotin ligase (BirA), which sequence-specifically ligates biotin to a 15 amino-acid acceptor peptide (AP). We report that BirA also accepts a ketone isostere of biotin as a cofactor, ligating this probe to the AP with similar kinetics and retaining the high substrate specificity of the native reaction. Because ketones are absent from native cell surfaces, AP-fused recombinant cell surface proteins can be tagged with the ketone probe and then specifically conjugated to hydrazide- or hydroxylamine-functionalized molecules. We demonstrate this two-stage protein labeling methodology on purified protein, in the context of mammalian cell lysate, and on epidermal growth factor receptor (EGFR) expressed on the surface of live HeLa cells. Both fluorescein and a benzophenone photoaffinity probe are incorporated, with total labeling times as short as 20 min. PMID- 15782207 TI - Gene identification signature (GIS) analysis for transcriptome characterization and genome annotation. AB - We have developed a DNA tag sequencing and mapping strategy called gene identification signature (GIS) analysis, in which 5' and 3' signatures of full length cDNAs are accurately extracted into paired-end ditags (PETs) that are concatenated for efficient sequencing and mapped to genome sequences to demarcate the transcription boundaries of every gene. GIS analysis is potentially 30-fold more efficient than standard cDNA sequencing approaches for transcriptome characterization. We demonstrated this approach with 116,252 PET sequences derived from mouse embryonic stem cells. Initial analysis of this dataset identified hundreds of previously uncharacterized transcripts, including alternative transcripts of known genes. We also uncovered several intergenically spliced and unusual fusion transcripts, one of which was confirmed as a trans splicing event and was differentially expressed. The concept of paired-end ditagging described here for transcriptome analysis can also be applied to whole genome analysis of cis-regulatory and other DNA elements and represents an important technological advance for genome annotation. PMID- 15782199 TI - Libraries enriched for alternatively spliced exons reveal splicing patterns in melanocytes and melanomas. AB - It is becoming increasingly clear that alternative splicing enables the complex development and homeostasis of higher organisms. To gain a better understanding of how splicing contributes to regulatory pathways, we have developed an alternative splicing library approach for the identification of alternatively spliced exons and their flanking regions by alternative splicing sequence enriched tags sequencing. Here, we have applied our approach to mouse melan-c melanocyte and B16-F10Y melanoma cell lines, in which 5,401 genes were found to be alternatively spliced. These genes include those encoding important regulatory factors such as cyclin D2, Ilk, MAPK12, MAPK14, RAB4, melastatin 1 and previously unidentified splicing events for 436 genes. Real-time PCR further identified cell line-specific exons for Tmc6, Abi1, Sorbs1, Ndel1 and Snx16. Thus, the ASL approach proved effective in identifying splicing events, which suggest that alternative splicing is important in melanoma development. PMID- 15782208 TI - Localized transfection on arrays of magnetic beads coated with PCR products. AB - High-throughput gene analysis would benefit from new approaches for delivering DNA or RNA into cells. Here we describe a simple system that allows any molecular biology laboratory to carry out multiple, parallel cell transfections on microscope coverslip arrays. By using magnetically defined positions and PCR product-coated paramagnetic beads, we achieved transfection in a variety of cell lines. Beads may be added to the cells at any time, allowing both spatial and temporal control of transfection. Because the beads may be coated with more than one gene construct, the method can be used to achieve cotransfection within single cells. Furthermore, PCR-generated mutants may be conveniently screened, bypassing cloning and plasmid purification steps. We illustrated the applicability of the method by screening combinatorial peptide libraries, fused to GFP, to identify previously unknown cellular localization motifs. In this way, we identified several localizing peptides, including structured localization signals based around the scaffold of a single C2H2 zinc finger. PMID- 15782209 TI - An extracellular matrix microarray for probing cellular differentiation. AB - We present an extracellular matrix (ECM) microarray platform for the culture of patterned cells atop combinatorial matrix mixtures. This platform enables the study of differentiation in response to a multitude of microenvironments in parallel. The fabrication process required only access to a standard robotic DNA spotter, off-the-shelf materials and 1,000 times less protein than conventional means of investigating cell-ECM interactions. To demonstrate its utility, we applied this platform to study the effects of 32 different combinations of five extracellular matrix molecules (collagen I, collagen III, collagen IV, laminin and fibronectin) on cellular differentiation in two contexts: maintenance of primary rat hepatocyte phenotype indicated by intracellular albumin staining and differentiation of mouse embryonic stem (ES) cells toward an early hepatic fate, indicated by expression of a beta-galactosidase reporter fused to the fetal liver specific gene, Ankrd17 (also known as gtar). Using this technique, we identified combinations of ECM that synergistically impacted both hepatocyte function and ES cell differentiation. This versatile technique can be easily adapted to other applications, as it is amenable to studying almost any insoluble microenvironmental cue in a combinatorial fashion and is compatible with several cell types. PMID- 15782212 TI - In vivo protein biotinylation for identification of organ-specific antigens accessible from the vasculature. AB - We describe a new methodology, based on terminal perfusion of rodents with a reactive ester derivative of biotin that enables the covalent modification of proteins readily accessible from the bloodstream. Biotinylated proteins from total organ extracts can be purified on streptavidin resin in the presence of strong detergents, digested on the resin and subjected to liquid chromatography tandem mass spectrometry for identification. In the present study, in vivo biotinylation procedure led to the identification of hundreds of proteins in different mouse organs, including some showing a restricted pattern of expression in certain body tissues. Furthermore, biotinylation of mice with F9 subcutaneous tumors or orthotopic kidney tumors revealed both quantitative and qualitative differences in the recovery of biotinylated proteins, as compared to normal tissues. This technology is applicable to proteomic investigations of the differential expression of accessible proteins in physiological and pathological processes in animal models, and to human surgical specimens using ex vivo perfusion procedures. PMID- 15782213 TI - Different delivery methods-different expression profiles. PMID- 15782214 TI - Measuring kinase activity: finding needles in a haystack. PMID- 15782215 TI - Herpesviruses throw a curve ball: new insights into microRNA biogenesis and evolution. PMID- 15782216 TI - In vivo protein labeling with trimethoprim conjugates: a flexible chemical tag. AB - The introduction of green fluorescent protein and its variants (GFPs) has allowed protein analysis at the level of the cell. Now, chemical methods are needed to label proteins in vivo with a wider variety of functionalities so that mechanistic questions about protein function in the complex cellular environment can be addressed. Here we demonstrate that trimethoprim derivatives can be used to selectively tag Escherichia coli dihydrofolate reductase (eDHFR) fusion proteins in wild-type mammalian cells with minimal background and fast kinetics. PMID- 15782217 TI - The hESC line Envy expresses high levels of GFP in all differentiated progeny. AB - Human embryonic stem cells (hESCs) have been advanced as a potential source of cells for use in cell replacement therapies. The ability to identify hESCs and their differentiated progeny readily in transplantation experiments will facilitate the analysis of hESC potential and function in vivo. We have generated a hESC line designated 'Envy', in which robust levels of green fluorescent protein (GFP) are expressed in stem cells and all differentiated progeny. PMID- 15782218 TI - Photo-leucine and photo-methionine allow identification of protein-protein interactions in living cells. AB - Protein-protein interactions are the key to organizing cellular processes in space and time. The only direct way to identify such interactions in their cellular environment is by photo-cross-linking. Here we present a new strategy for photo-cross-linking proteins in living cells. We designed two new photoactivatable amino acids that we termed photo-methionine and photo-leucine based on their structures and properties closely resembling the natural amino acids methionine and leucine, respectively. This similarity allows them to escape the stringent identity control mechanisms during protein synthesis and be incorporated into proteins by the unmodified mammalian translation machinery. Activation by ultraviolet light induces covalent cross-linking of the interacting proteins, which can be detected with high specificity by simple western blotting. Applying this technology to membrane protein complexes, we discovered a previously unknown direct interaction of the progesterone-binding membrane protein PGRMC1 with Insig-1, a key regulator of cholesterol homeostasis. PMID- 15782219 TI - Identification of microRNAs of the herpesvirus family. AB - Epstein-Barr virus (EBV or HHV4), a member of the human herpesvirus (HHV) family, has recently been shown to encode microRNAs (miRNAs). In contrast to most eukaryotic miRNAs, these viral miRNAs do not have close homologs in other viral genomes or in the genome of the human host. To identify other miRNA genes in pathogenic viruses, we combined a new miRNA gene prediction method with small-RNA cloning from several virus-infected cell types. We cloned ten miRNAs in the Kaposi sarcoma-associated virus (KSHV or HHV8), nine miRNAs in the mouse gammaherpesvirus 68 (MHV68) and nine miRNAs in the human cytomegalovirus (HCMV or HHV5). These miRNA genes are expressed individually or in clusters from either polymerase (pol) II or pol III promoters, and share no substantial sequence homology with one another or with the known human miRNAs. Generally, we predicted miRNAs in several large DNA viruses, and we could neither predict nor experimentally identify miRNAs in the genomes of small RNA viruses or retroviruses. PMID- 15782220 TI - A multiplexed homogeneous fluorescence-based assay for protein kinase activity in cell lysates. AB - New methods to quantify protein kinase activities directly from complex cellular mixtures are critical for understanding biological regulatory pathways. Herein, a fluorescence-based chemosensor strategy for the direct measurement of kinase activities in crude mammalian cell lysates is described. We first designed a new fluorescent peptide reporter substrate for each target kinase. These kinase chemosensors were readily phosphorylated by recombinant target enzyme and underwent a several-fold fluorescence increase upon phosphorylation. Then, using unfractionated cell lysates, a homogeneous kinase assay was developed that was reproducible, linear and highly preferential for monitoring changes in cellular activity of the target kinase. The general protocol was developed for the kinase Akt and then easily extended to measure protein kinase A (PKA) and mitogen activated protein kinase-associated protein kinase 2 (MK2) activities. This assay platform is immediately useful for studying protein kinase signaling in crude cellular extracts. PMID- 15782221 TI - Quantification of random genomic mutations. AB - Cancer cells contain numerous clonal mutations. It has been theorized that malignant cells sustain an elevated mutation rate and, as a consequence, harbor yet larger numbers of random point mutations. Testing this hypothesis has been precluded by lack of an assay to measure random mutations-that is, mutations that occur in only one or a few cells of a population. We have established a method that has permitted us to detect and identify rare random mutations in human cells, at a frequency of 1 per 10(8) base pairs. The assay is based on gene capture, by hybridization with a uracil-containing probe, followed by magnetic separation. Mutations that render the mutational target sequence non-cleavable by a restriction enzyme are quantified by dilution to single molecules and real-time quantitative PCR amplification. The assay can be extended to quantify mutation in any DNA-based organism, at different sites in the genome, in introns and exons, in unselected and selected genes, and in proliferating and quiescent cells. PMID- 15782224 TI - Sequencing drug response with HapMap. AB - The information about how DNA sequence varies across the human genome is crucial for unravelling the genetic basis of drug response. A haplotype map, or HapMap, intended to reveal such a variation pattern, has been recently developed by the International HapMap Consortium. Here, we present a conceptual model for directly characterizing specific DNA sequence variants that are responsible for drug response based on the haplotype structure provided by HapMap. Our model is developed in the maximum likelihood context, incorporated by clinically meaningful mathematical functions that model drug response and implemented with the EM algorithm. Our model is employed to a pharmacogenetic study of cardiovascular disease with 107 patients. We found that the haplotype constituted by allele Gly16 (G) at codon 16 and allele Glu27 (G) at codon 27 genotyped within the beta2AR candidate gene exhibits a different effect on heart rate curve from the rest haplotypes. Parents with the diplotype consisting of two copies of haplotype GG are more sensitive in heart rate to increasing dosages of dobutamine than those with other haplotypes. This model provides a powerful tool for elucidating the genetic variants of drug response and ultimately designing personalized medications based on each patient's genetic constitution. PMID- 15782225 TI - Ethnic differences in the relationships between obesity and glucose-metabolic abnormalities: a cross-sectional population-based study. AB - OBJECTIVES: To evaluate whether body mass index (BMI) and other anthropometric indices of visceral obesity vary by ethnic group in their distribution and their relationship to metabolic abnormalities. DESIGN: Cross-sectional study. PARTICIPANTS: Canadian men and women, aged 35-75 years, of South Asian (n=342), Chinese (n=317), European (n=326) and Aboriginal (n=301) descent were recruited using stratified random sampling. PRIMARY MEASURES: Anthropometric indices (BMI, waist to hip ratio (WHR) and waist circumference (WC)), metabolic markers (fasting glucose, HbA1c, the ratio of total cholesterol/HDL) and clinical markers (systolic blood pressure) were assessed. RESULTS: In subjects with BMI<30 kg/m2, the mean marker levels in people with elevated WC (>88 cm in women, >102 cm in men) vs people with normal WC were 6.16 vs 5.34 mmol/l for fasting glucose, 6.05 vs 5.66% for HbA1c and 5.46 vs 4.68 for the ratio of total cholesterol to HDL (P<0.001 in each case). At nearly every given level of BMI, non-European ethnic groups displayed significantly higher marker levels than Europeans. For example, for a given BMI, age and sex, the difference between European and non-European groups in HbA1c levels was 0.53% (95% confidence interval (CI): 0.37-0.69) for South Asians, 0.37% (95% CI: 0.2-0.54) for Chinese and 0.95% (95% CI: 0.78-1.12) for Aboriginal People. CONCLUSIONS: Uniform cut-points for the classification of obesity using BMI, WHR or WC result in marked variation in the levels of glucose metabolic abnormalities between ethnic groups. Existing action thresholds for these anthropometric indices do not apply to non-European ethnic groups and warrant revision. PMID- 15782226 TI - Body mass index and weight change from adolescence into adulthood, waist-to-hip ratio and perceived work ability among young adults. AB - OBJECTIVES: To study how body mass index (BMI, kg/m2) at 14 and 31 years (y) changes in BMI between 14 and 31 y, and waist-to-hip ratio (WHR) at 31 y are associated with poor perceived working ability at 31 y. DESIGN AND SUBJECTS: A population-based cohort, originally 11637 people, born in Northern Finland in 1966 was resurveyed at 14 and 31 y. MEASUREMENTS: Perceived work ability (measured by modified work ability index, WAI), BMI, WHR, alcohol intake, smoking, basic education and work history were recorded at 31 y and BMI also at 14 y. RESULTS: Low WAI (15% of lowest values) at 31 y showed a U-shaped association with BMI at 14 y, and also with BMI at 31 y, except in obese males. Low WAI similarly had a U-shaped association with WHR, but in males only, while in females, the probability of low WAI increased almost linearly with WHR. Low WAI was more common among smokers and people with a low level of education. The pattern for alcohol intake was more equivocal, with a nonsignificant finding in males and an almost linear decrease of low WAI with increasing alcohol intake in females. CONCLUSIONS: Work ability, health habits and anthropometric measures should be evaluated regularly in young workers, and preventive measures against diseases leading to premature retirement should be taken in time, for example by the occupational health-care service. PMID- 15782227 TI - Maternal weight status modulates the effects of restriction on daughters' eating and weight. AB - OBJECTIVE: To examine the effects of overweight and normal-weight mothers' restriction in child feeding on daughters' eating in the absence of hunger (EAH) and body mass index (BMI) change from age 5 to age 9 y. DESIGN: Longitudinal study of the health and development of young girls. SUBJECTS: A total of 91 overweight and 80 normal-weight mothers and their daughters, assessed when daughters were ages 5, 7, and 9 y. MEASUREMENTS: Measures included maternal restriction of daughters' intake at age 5 y, and daughters' EAH and BMI change from age 5 to 9 y. RESULTS: There were no overall differences in the level of restriction that overweight and normal-weight mothers used. However, overweight mothers' restrictive feeding practices when daughters were age 5 y predicted daughters' EAH over time, and higher EAH scores were associated with greater BMI change from age 5 to 9 y. These relationships did not hold for daughters of normal-weight mothers. CONCLUSION: More adverse effects of restriction on daughters' EAH, and links between EAH and BMI change were only noted among daughters of overweight mothers. These findings highlight the need for a better understanding of factors that contribute to within-group variation in eating behavior and weight status. PMID- 15782229 TI - Cadmium tolerance plasticity in Rhizobium leguminosarum bv. viciae: glutathione as a detoxifying agent. AB - Rhizobium leguminosarum bv. viciae strains expressing different degrees of tolerance to metal stress were used in this work to study the basic mechanisms underlying heavy metal tolerance. We used various parameters to evaluate this response. The strains' growth responses under different Cd2+ concentrations were determined and we reported variation in Cd2+ tolerance. Total soluble protein content decreased drastically, revealing the toxic effects that intracellular Cd2+ imposes on cellular metabolism, but this decrease in protein content was particularly evident in sensitive and moderately tolerant strains. Tolerant strains presented the highest intracellular and wall-bound Cd2+ concentrations. Cd2+ induced increases in the expression of some specific proteins, which were identical in all tolerant strains. Glutathione levels remained unaltered in the sensitive strain and increased significantly in tolerant and moderately tolerant strains, suggesting the importance of glutathione in coping with metal stress. This work suggests that efflux mechanisms may not be the only system responsible for dealing with heavy metal tolerance. A clear correlation between glutathione levels and Cd2+ tolerance is reported, thus adding a novel aspect in bacteria protection against heavy metal deleterious effects. PMID- 15782228 TI - Isolation of wood-inhabiting fungi from Canadian hardwood logs. AB - Wood-inhabiting fungi include many molds, wood-staining fungi, and decay fungi. Most of these fungal species can result in economic losses to wood users. Studies on molds, staining fungi, and decay fungi are necessary to be able to control their growth on wood and wood products. In this study, wood-inhabiting fungi were isolated from logs of 3 major Canadian hardwood species: sugar maple, white birch, and yellow birch. Two media were used for isolation. From these 3 wood species, a total of 1198 fungal cultures were obtained from summer- and winter harvested logs in dry storage and under water sprinkling. The results showed that most fungal species were not host specific and affected all of the wood species tested. Frequently isolated molds were Alternaria alternata, Trichoderma species, and Mucor/Rhizopus (Zygomycota) species, frequently isolated staining fungi were Ophiostoma piceae and Ophiostoma piliferum, a frequently isolated bark saprophyte was Nectria cinnabarina, and frequently isolated decay fungi were taxa of the phylum Basidiomycota. More fungal species were isolated from summer-harvested logs than from winter-harvested logs. Fewer fungal cultures, especially decay fungi, were isolated from logs in early storage than from logs in late storage. PMID- 15782230 TI - Characteristics of the rhizobia associated with Dalea spp. in the Ordway, Kellogg Weaver Dunes, and Hayden prairies. AB - Habitat fragmentation affects the biodiversity and function of aboveground organisms in natural ecosystems but has not been studied for effects on belowground species. In this paper, we consider the diversity of the rhizobia associated with the indigenous legume Dalea purpurea in 3 residual prairie areas in Minnesota and Iowa. Using Dalea purpurea as a trap host, 218 rhizobia were recovered from these soils then characterized using BOXA1R PCR. Three major and 13 minor groups were distinguished based on a similarity of greater than 75% in fingerprint patterns. Each major group consisted almost exclusively of rhizobia from a single prairie, with the diversity of Dalea rhizobia recovered from the Hayden Prairie less than that obtained with rhizobia from the other prairies. Based on 16S rRNA gene sequence analysis, isolates from the Hayden, Ordway, and Kellogg-Weaver Dunes prairies were most similar to Rhizobium etli and Rhizobium leguminosarum, Rhizobium gallicum, and Mesorhizobium amorphae and Mesorhizobium huakuii, respectively. This variation in the dominant microsymbiont species across the 3 prairies studied was unexpected but could have been influenced by the limited number of samples that we were allowed to take, by unanticipated cross-nodulation between native legumes, and by variation in the range of legume species present in each residual prairie area. While some of the rhizobia from Dalea nodulated Phaseolus vulgaris, Macroptilium atropurpureum, Leucaena leucocephala, and Onobrychis viciifolia in addition to the Dalea species tested, others nodulated Astragalus canadensis or Amorpha canescens. PMID- 15782231 TI - Prevalence and airborne spore levels of Stachybotrys spp. in 200 houses with water incursions in Houston, Texas. AB - Two hundred homes with a history of water incursion were sampled for fungi to determine the prevalence and airborne spore levels of Stachybotrys spp. Sampling methods included room air, surface, and wall cavity air sampling. Stachybotrys spp. were detected with at least one of the methods in 58.5% of the houses tested, but only 9.6% of the room air samples contained Stachybotrys spores. Aerosolization of Stachybotrys spores was correlated with both wall cavity and surface contamination. However, after adjustment for the surface effect, Stachybotrys spores detected in wall cavities were not a significant factor contributing to spores detected in room air samples. We conclude that Stachybotrys spp. are commonly found on water-damaged building materials. In addition, the observations made in this study suggest that the impact on the living space air is low if the fungal spores are contained within a wall cavity. PMID- 15782232 TI - Genome-wide comparison of phage M13-infected vs. uninfected Escherichia coli. AB - To identify Escherichia coli genes potentially regulated by filamentous phage infection, we used oligonucleotide microarrays. Genome-wide comparison of phage M13-infected and uninfected E. coli, 2 and 20 min after infection, was performed. The analysis revealed altered transcription levels of 12 E. coli genes in response to phage infection, and the observed regulation of phage genes correlated with the known in vivo pattern of M13 mRNA species. Ten of the 12 host genes affected could be grouped into 3 different categories based on cellular function, suggesting a coordinated response. The significantly upregulated genes encode proteins involved in reactions of the energy-generating phosphotransferase system and transcription processing, which could be related to phage transcription. No genes belonging to any known E. coli stress response pathways were scored as upregulated. Furthermore, phage infection led to significant downregulation of transcripts of the bacterial genes gadA, gadB, hdeA, gadE, slp, and crl. These downregulated genes are normally part of the host stress response mechanisms that protect the bacterium during conditions of acid stress and stationary phase transition. The phage-infected cells demonstrated impaired function of the oxidative and the glutamate-dependent acid resistance systems. Thus, global transcriptional analysis and functional analysis revealed previously unknown host responses to filamentous phage infection. PMID- 15782233 TI - Polarity within pM and pE promoted phage lambda cI-rexA-rexB transcription and its suppression. AB - The cI-rexA-rexB operon of bacteriophage lambda confers 2 phenotypes, Imm and Rex, to lysogenic cells. Immunity to homoimmune infecting lambda phage depends upon the CI repressor. Rex exclusion of T4rII mutants requires RexA and RexB proteins. Both Imm and Rex share temperature-sensitive conditional phenotypes when expressed from cI[Ts]857 but not from cI+ lambda prophage. Plasmids were made in which cI-rexA-rexB was transcribed from a non-lambda promoter, pTet. The cI857-rexA-rexB plasmid exhibited Ts conditional Rex and CI phenotypes; the cI+ rexA-rexB plasmid did not. Polarity was observed within cI-rexA-rexB transcription at sites in cI and rexA when CI was nonfunctional. Renaturation of the Ts CI857 repressor, allowing it to regain functionality, suppressed the polar effect on downstream transcription from the site in cI. The second strong polar effect near the distal end of rexA was observed for transcription initiated from pE. The introduction of a rho Ts mutation into the host genome suppressed both polar effects, as measured by its suppression of the conditional Rex phenotype. Strong suppression of the conditional Rex[Ts] phenotype was imparted by ssrA and clpP (polar for clpX) null mutations, suggesting that RexA or RexB proteins made under conditions of polarity are subject to 10Sa RNA tagging and ClpXP degradation. PMID- 15782234 TI - Antimicrobial susceptibility of lactic acid bacteria isolated from a cheese environment. AB - In the production of the Spanish traditional blue-veined Cabrales cheese, lactic acid bacteria strains free of antibiotic resistance that have a transferrable capacity are necessary as components of a specific starter. To select for these bacteria, the minimum inhibitory concentration (MIC) of 12 antibiotics and 2 mixtures (containing beta-lactamase inhibitor and penicillin) were determined by microbroth and agar dilution techniques in 146 strains belonging to the genera Lactococcus, Enterococcus, Lactobacillus, and Leuconostoc. The antibiotic resistance profiles of Lactococcus and Enterococcus species were different from those of Lactobacillus and Leuconostoc, but clear genus- or species-associated patterns were not observed. Cefoxitin and metronidazole were not effective against bacteria of these genera. The MICs of beta-lactam antibiotics for lactobacilli and leuconostoc isolates were higher than those for lactococci and enterococci, but no strain was clinically resistant. All lactobacilli and leuconostoc isolates were resistant to high levels of vancomycin, a type of resistance not seen among the tested members of the genera Lactococcus and Enterococcus. The majority of the observed resistance appeared to be either intrinsic or nonspecific, although some strains of Lactococcus lactis, Enterococcus spp., and Lactobacillus spp. were resistant to antibiotics, such as chloramphenicol, erythromycin, clindamycin, or tetracycline. PMID- 15782235 TI - Characterization and regulation of new secondary metabolites from Aspergillus ochraceus M18 obtained by UV mutagenesis. AB - UV irradiation of Aspergillus ochraceus NRRL 3174 conidia led to stable mutations in ochratoxin and penicillic-acid pathways. These mutants, especially M18, produced an unexpectedly large number of new metabolites. Two new compounds were purified by TLC and HPLC and their chemical structures were determined. They are 2,10-dimethyl 4-hydroxy-6-oxo-4-undecen-7-yne (1) and 4-(3-methyl-2- butenyl) oxy 1-phenyl acetic acid (2). Compound 1 is very active against Gram-positive bacteria, such as Staphylococcus aureus and Bacillus subtilis, but inactive against Gram-negative bacteria, fungi, and yeasts. However, compound 2 has no antibiotic activity. The production of 1 was generally associated with growth, whereas that of compound 2 was dissociated from growth. The biosynthesis of these 2 metabolites was influenced by the sources of carbon and nitrogen. PMID- 15782236 TI - [Effect of aeration on composting of date palm residues contaminated with Fusarium oxysporum f.sp. albedinis]. AB - Composting of date palm (Phoenix dactylifera L.) residues contaminated with Fusarium f.sp oxysporum albedinis, causal agent of the vascular wilt (Bayoud) of the date palm, has been achieved. The effect of the aeration of the piles by manual turning has been studied. The maintenance of an adequate humidity of 60% 70%, necessary to the good progress of the composting process, required the contribution of 11.4 L of water/kg of the dried residues. The evolution of the temperatures in the three piles presents the same phases. A latency phase, followed after 2-3 d of composting by a thermophilic phase, which lasts about 24 d, where the temperature remains elevated between 50 and 70 degrees C. Then a cooling phase that takes about 15 d, during which the temperatures fall to values between 25 and 35 degrees C, near room temperature. Fusarium f.sp oxysporum albedinis is eliminated completely during the thermophilic phase of composting, and increasing frequencies of turning accelerate its disappearance to a certain extent. On the other hand, pH remained steady and relatively basic oscillating between 8.2 and 8.7. Ninety percent (90%) of the the date palm residues are composed exclusively of organic matters. The total nitrogen represents only 0.4%. The contribution of manure decreases the ratio of carbon to nitrogen (C/N) from 115 to 48 in the initial mixture. After 80 d of composting and according to the frequency of return up, there is a reduction of the granulometry of the substratum, the C/N ratio (from 29% to 44%), the organic matter (from 15% to 23%), the total volume (from 25% to 35%), and of the dry weight of the swaths (from 16% to 24%). On the other hand there is an increase in total nitrogen rate (from 20% to 40%) and in the mineral matter (from 23% to 35%). PMID- 15782237 TI - [Potential activity of methane production in soil, peat, and lacustrine sediments and in the Robert Bourassa hydro-electric reservoir in northern Canada]. AB - Flooding of land associated with the creation of reservoirs may increase, at least in the short term, methane flux to the atmosphere. To evaluate the potential contribution of such land use on methane production, field samples were studied in vitro for the potential activity of methanogenic bacteria in unflooded or flooded boreal forest soils, together with lacustrine sediments. From this comparative study, periodically flooded or flooded peats contribute more to methane production than do unflooded peats, soils, and natural lake sediment. The intensity and temporal changes in the activity of methanogenic archaea in the different systems depended on a combination of environmental factors, such as the amount and quality of organic carbon, the water level, and the concentration of oxidizing ions (SO42-, Fe3+). PMID- 15782239 TI - Influence of carbon source and surface hydrophobicity on the aggregation of the yeast Kluyveromyces bulgaricus. AB - Aggregation of the yeast Kluyveromyces bulgaricus is mediated by the galactose specific lectin KbCWL1. This lectin contains hydrophobic amino acids and its activity is calcium dependent. A specific fluorescent probe, 1-anilinonaphthalene 8-sulfonic acid in the free acid form (ANS; Sigma Chemical Co., St. Louis, Missouri), was used to study the hydrophobic areas on the cellular surface of K. bulgaricus. Changes in surface hydrophobicity during the growth and aggregation of yeast cells were studied. Surface hydrophobicity increased during growth and depended on the amount of yeast cells in the culture medium. During growth, the size of the hydrophobic areas on the cell surface was measured using ANS and was found to increase with the percentage of flocculating yeasts. Our results strongly suggest that the hydrophobic areas of the cell walls of yeast cells are involved in the aggregation of K. bulgaricus. PMID- 15782238 TI - Overexpression of the ftsZ gene from Corynebacterium glutamicum (Brevibacterium lactofermentum) in Escherichia coli. AB - Our goal in this work was to overexpress the essential cell division FtsZ protein from Corynebacterium glutamicum (Brevibacterium lactofermentum) (FtsZCG) in Escherichia coli to produce anti-FtsZCG polyclonal antibodies. Previous results from our laboratory showed that ftsZCG was not expressed in E. coli in a sufficient amount to purify FtsZCG. However, when ftsZCG (without upstream sequences) was transcriptionally fused to the T7 promoter, different truncated FtsZCG proteins (28-32 kDa) were overexpressed in E. coli, and in all cases, stop codons were created because of DNA deletions or rearrangements. Nevertheless, we were able to overexpress and purify an N-terminally hexa-His-tagged FtsZCG from uninduced E. coli cells carrying a pET-28a(+) derivative, yielding about 5 mg of 98% pure protein per 100-mL culture. PMID- 15782240 TI - Isolation and PCR amplification of a species-specific oxidoreductase-coding gene region in Listeria grayi. AB - Listeria grayi is a nonpathogenic Gram-positive bacterium that demonstrates considerable similarities to other members in the genus Listeria, including the foodborne human pathogen Listeria monocytogenes and the animal pathogen Listeria ivanovii. A rapid diagnostic test to identify and diagnose listeriosis would be valuable, especially in cases where the presence of L. grayi may complicate diagnosis. This test would be based on a unique gene present in L. grayi. In this study, after comparative screening of a recombinant L. grayi DNA library by dot blot hybridization, an L. grayi specific clone (lgr20-246) with an insert of 722 bp was isolated. By applying PCR primers derived from a distinct region of the clone not shared by other bacteria, a specific band of 420 bp was amplified from the genomic DNA of L. grayi only and not of other Listeria species or common bacteria. These results suggest that the PCR assay employing primers lgr20-246F and lgr20-246R provides an independent and precise means of distinguishing L. grayi from other Listeria species and common bacteria. Therefore, it would be another useful technique for laboratory differentiation of Listeria bacteria. PMID- 15782241 TI - The presence of novel 3-hydroxy oxylipins on surfaces of hat-shaped ascospores of Ascoidea africana Batra & Francke-Grosmann. AB - Immunofluorescence microscopy exposed the presence of novel 3-hydroxy oxylipins on the surfaces of aggregated hat-shaped ascospores of Ascoidea africana. These compounds were confirmed by gas chromatography - mass spectrometry analysis. Only the complete structure of a novel 3-hydroxy 10:1 could be determined. PMID- 15782242 TI - Vagus nerve stimulation affects pain perception in depressed adults. AB - BACKGROUND: Previous research suggests that vagus nerve stimulation (VNS) affects pain perception in epilepsy patients, with acute VNS decreasing pain thresholds and chronic VNS treatment increasing pain thresholds. However, no studies have investigated the effects of VNS on pain perception in chronically depressed adults, nor have controlled, systematic investigations been published on the differential effects of certain VNS device parameters on pain perception. OBJECTIVES: The present study tried to replicate the results of previous research showing acute pronociceptive effects of VNS and determine the effects of various device parameter settings on pain tolerance. The present study also investigated the relationship among patients' levels of depression, duration of VNS treatment and VNS-induced changes in pain perception. METHODS: A thermal pain challenge task was used to determine pain tolerance during VNS device activation using different combinations of VNS device parameter settings within subjects undergoing VNS therapy for chronic depression. RESULTS: Significant pronociceptive effects were found for acute VNS activation. Individual differences were found with respect to the VNS settings associated with the largest changes in pain perception. Severity of depression was inversely related to baseline pain tolerance, but depression severity was unrelated to VNS-induced acute changes in pain tolerance, as was the length of time participants had been undergoing VNS treatment. CONCLUSIONS: VNS appears to affect pain perception in depressed adults. Different VNS parameter settings may be associated with unique effects from patient to patient. More studies are needed to determine the long term effects of VNS on pain perception. PMID- 15782243 TI - The face of pain--a pilot study to validate the measurement of facial pain expression with an improved electromyogram method. AB - OBJECTIVE: The purpose of this pilot study was to establish the validity of an improved facial electromyogram (EMG) method for the measurement of facial pain expression. BACKGROUND: Darwin defined pain in connection with fear as a simultaneous occurrence of eye staring, brow contraction and teeth chattering. Prkachin was the first to use the video-based Facial Action Coding System to measure facial expressions while using four different types of pain triggers, identifying a group of facial muscles around the eyes. METHOD: The activity of nine facial muscles in 10 healthy male subjects was analyzed. Pain was induced through a laser system with a randomized sequence of different intensities. Muscle activity was measured with a new, highly sensitive and selective facial EMG. RESULTS: The results indicate two groups of muscles as key for pain expression. These results are in concordance with Darwin's definition. As in Prkachin's findings, one muscle group is assembled around the orbicularis oculi muscle, initiating eye staring. The second group consists of the mentalis and depressor anguli oris muscles, which trigger mouth movements. CONCLUSIONS: The results demonstrate the validity of the facial EMG method for measuring facial pain expression. Further studies with psychometric measurements, a larger sample size and a female test group should be conducted. PMID- 15782244 TI - Treatment of whiplash-associated disorders--part I: Non-invasive interventions. AB - BACKGROUND: A whiplash-associated disorder (WAD) is an injury due to an acceleration-deceleration mechanism at the neck. WAD represents a very common and costly condition, both economically and socially. In 1995, the Quebec Task Force published a report that contained evidence-based recommendations regarding the treatment of WAD based on studies completed before 1993 and consensus-based recommendations. OBJECTIVE: The objective of the present article--the first installment of a two-part series on interventions for WAD--is to provide a systematic review of the literature published between January 1993 and July 2003 on noninvasive interventions for WAD using meta-analytical techniques. METHODS OF THE REVIEW: Three medical literature databases were searched for identification of all studies on the treatment of WAD. Randomized controlled trials (RCTs) and epidemiological studies were categorized by treatment modality and analyzed by outcome measure. The methodological quality of the RCTs was assessed. When possible, pooled analyses of the RCTs were completed for meta-analyses of the data. The results of all the studies were compiled and systematically reviewed. RESULTS: Studies were categorized as exercise alone, multimodal intervention with exercise, mobilization, strength training, pulsed magnetic field treatment and chiropractic manipulation. A total of eight RCTs and 10 non-RCTs were evaluated. The mean score of methodological quality of the RCTs was five out of 10. Pooled analyses were completed across all treatment modalities and outcome measures. The outcomes of each study were summarized in tables. CONCLUSIONS: There exists consistent evidence (published in two RCTs) in support of mobilization as an effective noninvasive intervention for acute WAD. Two RCTs also reported consistent evidence that exercise alone does not improve range of motion in patients with acute WAD. One RCT reported improvements in pain and range of motion in patients with WAD of undefined duration who underwent pulsed electromagnetic field treatment. Conflicting evidence in two RCTs exists regarding the effectiveness of multimodal intervention with exercise. Limited evidence, in the form of three non-RCTs, exists in support of chiropractic manipulation. Future research should be directed toward clarifying the role of exercise and manipulation in the treatment of WAD, and supporting or refuting the benefit of pulsed electromagnetic field treatment. Mobilization is recommended for the treatment of pain and compromised cervical range of motion in the acute WAD patient. PMID- 15782245 TI - Treatment of whiplash-associated disorders--part II: Medical and surgical interventions. AB - BACKGROUND: Whiplash-associated disorder (WAD) is a term used to describe injury due to an acceleration-deceleration mechanism at the neck. In 1995, the Quebec Task Force published a report that contained evidence-based recommendations regarding the treatment of WAD based on studies completed before 1993 and consensus-based recommendations. OBJECTIVE: The objective of the present article- the second installment of a two part review on WAD--is to provide a systematic review of the literature published between January 1993 and July 2003 on medical and surgical interventions for WAD using meta-analytical techniques. METHODS OF THE REVIEW: Three medical literature databases were searched for identification of all clinical trials on the treatment of WAD. Randomized controlled trials (RCTs) and epidemiological studies were categorized by treatment modality and analyzed by outcome measures. The methodological quality of the RCTs was assessed. When possible, pooled analyses of the RCTs were completed for meta analyses of the data. The results of all the studies were compiled and systematically reviewed. RESULTS: The studies were categorized as medical or surgical interventions. A total of five RCTs and six non-RCTs were evaluated. The mean methodological quality of the RCTs was 7.3 among studies on medical interventions and 7.5 for studies on surgical intervention. Pooled analyses were completed across all treatment modalities and outcome measures. The outcomes of each study were summarized in tables. CONCLUSIONS: While several quality RCTs and epidemiological studies have been published since 1993 on medical and surgical interventions, the cumulative evidence is lacking. Moderate evidence exists in support of radiofrequency neurotomy. Evidence for steroid injections, botulinum treatments, carpal tunnel decompression and cervical discectomy is conflicting or unclear. Future research is required to clarify the utility of radiofrequency neurotomy and pulsed electromagnetic field treatment for WAD. PMID- 15782246 TI - Ventrolateral partial dorsal root entry zone rhizotomy for occipital neuralgia. AB - OBJECTIVE AND IMPORTANCE: Medically refractory occipital neuralgia (ON) has been treated with a variety of neuroablative procedures. The present case report supports the effectiveness of ventrolateral partial rhizotomy (pVL-DREZ) of the C1 to C3 cervical dorsal roots, a relatively unknown procedure. CLINICAL PRESENTATION: A 46-year-old woman had a 14-month history of severe right-sided ON. Multiple trials of medical treatments, nerve blocks and local steroid injections had failed. Her daily opioid requirements had escalated to include frequent injections in addition to prescribed oral opiates. INTERVENTION AND RESULTS: A pVL-DREZ at the right C1 to C3 level was performed through a C1 laminectomy and C2 right laminotomy. The ipsilateral upper cervical dorsal roots were exposed and a 1 mm deep incision was made at a 45 degrees angle to the sagittal midline at the ventrolateral aspect of each dorsal rootlet entry. The patient experienced postoperative opioid withdrawal seizure and transient disequilibrium for two weeks. Touch sensation was preserved and complete abolition of ON over a four year follow-up was achieved. CONCLUSION: The pVL-DREZ procedure provided complete pain relief for the patient and avoided the potential complications often encountered with other destructive interventions. pVL-DREZ should be considered among the available options for the treatment of refractory ON. PMID- 15782247 TI - Structural studies of group 1 metal 4-azapentalenyl complexes. AB - The first structurally authenticated 4-azapentalenyl complexes are reported, along with DFT studies, that reveal tendencies for their cyclopentadienyl isostructural nature and an ability to exhibit binding modes of varying hapticity. PMID- 15782248 TI - A ferrocene-based heteroditopic ligand for electrochemical sensing of cations and anions. AB - A ferrocene-based heteroditopic receptor containing urea and crown ether units shows electrochemical responses to dihydrogenphosphate and fluoride anions. K+ cations can only be detected in the presence of dihydrogenphosphate. PMID- 15782249 TI - A step forward in gold-silver metallophilicity. An AuAg4 moiety with a square pyramidal arrangement. AB - By reaction of the Lewis base NBu4[Au(3,5-C6F3Cl2)2] and AgCF3COO in the presence of NBu4CF3COO the heterometallic compound (NBu4)2[Au(3,5-C6F3Cl22Ag4(CF3COO)5] (2) is obtained. The structure displays an unprecendented square pyramidal AuAg4 arrangement built up through four AuAg closed-shell interactions and two Au-C-Ag 3c-2e- bridges. PMID- 15782250 TI - Rational design and construction of the first tetrahedral net with photoluminescent Cu4I4 cubane cluster as the tetrahedral node. AB - Solvothermal reaction of CuI with flexible 1,3-bis(4-pyridyl)propane generated the first three-dimensional coordination polymer constructed with the rigid tetrahedrally connected Cu4I4 cluster unit. The net is a rare chiral triple interpenetrated, quartz net with of vertex symbol 6(4).8(2). The solid-state luminescence spectrum displays a strong red emission band at room temperature (lambdamax= 601 nm), characteristic of the Cu4I4 cluster centers. PMID- 15782251 TI - Hairpin helicates: a missing link between double-helicates and trefoil knots. AB - We have prepared a novel ligand in which two qpy metal-binding domains are linked by a polyoxyethylene spacer. The new ligand reacts with copper(ii) salts to form dinuclear double helicates with a hairpin structure. The hairpin complex and a model complex have been structurally characterised. The hairpin structure is formed regioselectively as the head-to-tail (HT) stereoisomer. Detailed electrochemical and spectroelectrochemical studies of the complexes are reported. PMID- 15782252 TI - Synthesis, characterization, in vitro antitumor activity, DNA-binding properties and electronic structure (DFT) of the new complex cis (Cl,Cl)[RuIICl2(NO+)(terpy)]Cl. AB - The complex cis-(Cl,Cl)-[RuCl2(terpy)(NO)]Cl (1) has been synthesized by the reaction of [RuCl3(H2O)2(NO)] with terpyridine (terpy) and characterized by various spectroscopic, analytical techniques and using electronic structure calculation (DFT) methods. The cytotoxic activity and the DNA-binding properties of have also been studied using biochemical techniques. The results establish unequivocally that corresponds to a so-called [RuNO]6 species, which readily releases the nitrosyl ligand upon irradiation with a mercury lamp in acetonitrile solution. DFT calculations provided a satisfactory description of structural, bonding, electronic and related properties of the new compound and throw light on the mechanism of the photo-induced NO release. Screening on A2780 (human ovarian carcinoma) cell lines showed significant cytotoxicity with an IC50 value of 0.49 microM. 31P and 23Na NMR spectroscopy along with electrophoretic mobility studies illustrated that complex primarily binds by coordination to DNA without any pi interaction between the planar terpy ligand and the DNA bases, while weak electrostatic interactions could not be excluded. Studies on the inhibition of the restriction enzymes DraI and SmaI revealed that prefers the guanine and cytosine bases of DNA. PMID- 15782253 TI - Isomeric ruthenium terpyridine complexes [Ru(trpy)(L)Cl]n+ containing the unsymmetrically bidentate acceptor L=3-amino-6-(3,5-dimethylpyrazol-1-yl)-1,2,4,5 tetrazine. Synthesis, structures, electrochemistry, spectroscopy and DFT calculations. AB - The isomeric title complexes were obtained in almost equimolar ratio from the reaction of Ru(trpy)Cl3 and L. Crystal structure analyses of the perchlorate hemihydrates, electrochemical and spectroscopic (NMR, UV/VIS, EPR) studies, supported by DFT calculations, reveal distinct differences between the isomeric redox series [1]n+(tetrazine-Nt trans to Cl) and [2]n+(pyrazolyl-Np trans to Cl; n= 0, 1, 2). The latter system with the pi acceptors trpy and tetrazine in the equatorial plane and the pyrazolyl and chloride donors in the axial positions exhibits facilitated oxidation, lower energy MLCT transitions, more balanced chelate coordination, and a higher g anisotropy in the oxidised (RuIII) state. According to partially resolved EPR spectra of one-electron reduced neutral compounds and they have the unpaired electron predominantly in the tetrazine ring of L. PMID- 15782254 TI - Alkoxo-bridged copperII complexes as nodes in designing solid-state architectures. The interplay of coordinative and d10-d10 metal-metal interactions in sustaining supramolecular solid-state architectures. AB - Four novel polymeric coordination networks have been obtained through self assembly processes involving alkoxo-bridged copperII species as nodes, and anionic cyano-complexes as linkers: infinity2[{Cu2(pa)2}{M(CN)2}2](M=Ag, 1; Au, 2), (infinity)3[{Cu4(mea)4}{Au(CN)2}4.H2O]3, and (infinity)3[{Cu2(pa)2}{Ni(CN)4}](pa = deprotonated propanolamine; mea = deprotonated monoethanolamine). The supramolecular architectures of compounds 1, and 2 are sustained by argentophilic or strong aurophilic interactions. The solid state architectures of 1 and 2, which are isomorphous, consist of infinite layers, constructed from binuclear alkoxo-bridged nodes and [M(CN)2]- spacers. The layers are stacked in an offset parallel mode, and are further interconnected through Ag...Ag or Au...Au contacts (1: Ag...Ag 3.015 A; 2: Au....Au 3.069 A). Compound 3 consists of unique fourfold interpenetrating diamondoid nets. The diamondoid topology is built of heterocubane {Cu4O4} nodes, which are connected by [Au(CN)2]- rods. The Cu-O distances within the {Cu4O4} node vary between 1.927(2) and 2.679(1) A, showing unsymmetric bridging of the copper atoms. Aurophilic interactions are established between the bridging and terminal [Au(CN)2]- metalloligands, and connect the interpenetrating nets, resulting in infinite chains of gold atoms (the Au...Au distances vary between 3.253 and 3.305 [Angstrom]). Compound 4 exhibits a 3-D network constructed from {Cu2(pa)2]2+ nodes connected by square-planar [Ni(CN)4]2- ions. Compounds 1, 2 and 4 are weakly paramagnetic. The cryomagnetic investigation of reveals a gradual increase, followed by a decrease of the chiMT product, as the temperature is lowered. A superposition of ferro- (J1=+20.8 cm(-1)) and antiferromagnetic (J2= 6.4) interactions within the tetranuclear node was found. Antiferromagnetic interactions are established between the tetranuclear nodes (theta=-2.99 K). PMID- 15782255 TI - Preparation of stable AsBr4+ and I2As-PI3+ salts. Why didn't we succeed to prepare AsI4+ and As2X5+? A combined experimental and theoretical study. AB - In analogy to our successful "PX2+" insertion reactions, an "AsX2+" insertion route was explored to obtain new arsenic halogen cations. Two new salts were prepared: AsBr4+[Al(OR)4]-, starting from AsBr3, Br2 and Ag[Al(OR)4], and I2As PI3+[Al(OR)]4 from AsI3, PI3 and Ag[Al(OR)4](R=C(CF3)3). The first cation is formally a product of an "AsBr2+" insertion into the Br2 molecule and the latter clearly a "PI2+" insertion into the As-I bond of the AsI3 molecule. Both compounds were characterized by IR and NMR spectroscopy, the first also by its X ray structure. Reactions of Ag[Al(OR)4] with AsI3 do not lead to ionization and AgI formation but rather lead to a marginally stable Ag(AsI3)2+[Al(OR)]4 salt. Despite many attempts we failed to prepare other PX-cation analogues such as AsI4+, As2X5+ and P4AsX2+(X=Br, I). To explain these negative results the thermodynamics of the formation of EX2+, EX4+ and E2X5+(E = As, P; X = Br, I) was carefully analyzed with MP2/TZVPP calculations and inclusion of entropy and solvation effects. We show that As2Br5+ is in very rapid equilibrium with AsBr2+ and AsBr3(DeltaGo((CH2Cl2))=+30 kJ mol(-1)). The extremely reactive AsBr2+ cation available in the equilibrium accounts for the observed decomposition of the [Al(OR)4]- anion. By contrast, the stability of AsI3 against Ag[Al(OR)4] appears to be kinetic and, if prepared by a suitable route, As2I5+ would be expected to have a stability intermediate between the known P2I5+ and P2Br5+. PMID- 15782256 TI - Electronic structure and ionization energies of palladium and platinum N heterocyclic carbene complexes. AB - Density functional methods have been used to calculate the geometries, electronic structure and ionization energies (IE) of N-heterocyclic carbene complexes of palladium and platinum, [M(CN2R2C2H2)2](M = Pd, Pt; R = H, Me, Bu t). Agreement with X-ray structures (R = Bu t) was good. Calculated IE agreed well with the photoelectron (PE) spectra (R = Bu t); metal bands were calculated to be within 0.25 eV of the experimental values, whereas the higher lying ligand bands deviated by up to 0.9 eV. Spin-orbit methods were needed to achieve this level of agreement for the Pt complex, but the calculations were found to underestimate the spin-orbit splitting somewhat. The principal metal-ligand bonding is between the carbene lone pair HOMO and a (d(z2)+ s) hybrid on the metal. The metal p(z) orbital contributes very little to the bonding. The metal d(xz,yz) orbitals mix primarily with the filled pi3 orbitals on the ligands and secondarily with the empty pi5 orbitals. Consequently they are little stabilized in comparison to the metal d(xy,x2- y2) orbitals, which are non-bonding in the complex. The first PE band for both the Pd and Pt complexes is from ionization of a (s - d(z2)) hybrid orbital. The IE is greater for Pt than for Pd on account of the post-lanthanide relativistic stabilization of the Pt 6s orbital. PMID- 15782257 TI - A combination of access to preassociation sites and local accumulation tendency in the direct vicinity of G-N7 controls the rate of platination of single stranded DNA. AB - Adduct formation between cationic reagents and targets on DNA are facilitated by the ability of DNA to attract cations to its surface. The electrostatic interactions likely provide the basis for the documented preference exhibited by cisplatin and related compounds for nuclear DNA over other cellular constituents. As an extension of a previous communication, we here present an investigation illustrating how the rate of adduct formation with the naturally occurring base guanine (G-N7) can be modulated by i) bulk solvent conditions, ii) local nature and size of the surrounding DNA and, iii) increasing DNA concentration. A series of single-stranded DNA oligomers of the type d(TnGTm); n= 0, 2, 4, 6, 8, 10, 12, 14, 16 and m= 16 -n or n=m= 4, 6, 8, 12, 16, 24 were allowed to react with the active metabolite of a potential orally active platinumIV drug, cis-[PtCl(NH3))(c C6H11NH2)(OH2)]+ in the presence of three different bulk cations; Na+, Mg2+, and Mn2+. For all positions along the oligomers, a change from monovalent bulk cations to divalent ones results in a decrease in reactivity, with Mn2+ as the more potent inhibitor as exemplified by the rate constants determined for interaction with d(T8GT8): 10(3) x k obs/s(-1)= 6.5 +/- 0.1 (Na+), 1.8 +/- 0.1 (Mg2+), 1.0 +/- 0.1 (Mn2+) at pH 4.2 and 25 degrees C. Further, the adduct formation rate was found to vary with the exact location of the binding site in the presence of both Na+ and Mg2+, giving rise to reactivity maxima at the middle position. Increasing the size of the DNA-fragments was found to increase the reactivity only up to a total length of ca. 20 bases. The influence from addition of further bases to the reacting DNA was found to be salt dependent. At [Na+]= 0.5 mM a retardation in reactivity was observed whereas [Na+] < or = 4.5 mM give rise to length independent kinetics. Finally, for the first time we have here been able to evaluate the influence from an increasing concentration of non reactive DNA bases on the adduct formation process. The latter data were successfully fitted to an inhibition model suggesting that non-productive association of the platinum complex with sites distant from G-N7 competes with productive ones in the vicinity of the G-N7 target. Taken together, the kinetics support a reaction mechanism in which access to suitable association sites in the direct vicinity of the target site controls the rate of platination. PMID- 15782258 TI - Synthesis and reactivity studies of a manganese 'microperoxidase' containing b type heme. AB - Mn(III)protoporphyrin IX-6(7)-gly-gly-his methyl ester (MnGGH) has been prepared by condensation of glycyl-glycyl-L-histidine methyl ester with the propionic side chains of Mn(III)protoporphyrin IX. It was characterised by mass spectrometry and UV/VIS spectroscopy. Stopped-flow spectrophotometry was used to study the reaction of the Mn 'microperoxidase' with hydrogen peroxide. The formation of active intermediates analogous to previously described metal-hydroperoxo (compound 0) and metal-oxo (compound I) intermediates of the 'natural' Fe(III) microperoxidase-8 and Mn(III) microperoxidase-8 was observed. The rate of formation of the MnGGH-based compound I analogue was found to increase dramatically with increasing pH. A steady-state kinetic analysis of the catalytic peroxidase activity of MnGGH towards K4[Fe(CN)6], L-tyrosine methyl ester, o dianisidine, o-methoxyphenol and ascorbic acid showed that the peroxidase reaction proceeds via the formation of a microperoxidase-substrate complex followed by electron transfer from the substrate to the metal. The reactivity of MnGGH depends on the size and hydrophobicity of the substrate, and these properties appear to influence the rate of the electron transfer, which is the rate-limiting step for the whole process. MnGGH showed higher reactivity towards reducing substrates than its Fe(iii) analogue. PMID- 15782259 TI - Synthesis and structure of the organometallic MFe2(mu3-S)2 clusters (M = Mo or Fe). AB - New organometallic clusters with the MFe2(mu3-S)2 core (M = Mo or Fe) have been synthesized from inorganic [MoFe3S4] or [Fe4S4] clusters under high pressure CO. The reaction of (Cl4-cat)2Mo2Fe6S8(PR3)6[R = Et, (n)Pr] with high pressure CO produced the crystalline [MoFe2S2]4+ clusters, (Cl4-cat)Mo(O)Fe2S2(CO)(n)(PR3)6 n[n= 4, Et =I, (n)Pr =II; n = 5, Et =III] after flash column chromatography. The similar [MoFe2S2]4+ cluster, (Cl4-cat)2MoFe2S2(CO)2(depe)(2)(IV), also has been achieved by the reactions of (Cl4-cat)MoFe3S3(CO)6(PEt3)2 with depe by reductive decoupling of the cluster. For the [Fe3(mu3-S)2]4+ cluster, [Fe4S4(PcHex3)4](BPh4) was reacted with high pressure CO to produce a new Fe3S2(CO)7(PcHex)(2)(V) compound. These reactions generalized the preparation of organometallic compounds from inorganic clusters. All the compounds have been characterized by single crystal X-ray crystallography. A possible reaction pathway for the synthesis of the MFe2(mu3-S) clusters (M = Mo or Fe) has also been suggested. PMID- 15782260 TI - Synthesis and electrochemical studies of a covalently linked cobalt(II) phthalocyanine-cobalt(II) porphyrin conjugate. AB - The synthesis of a cobalt phthalocyanine-cobalt porphyrin heteropentamer (cobalt(II) phthalocyanine-tetrakis(cobalt(II) tetrakis(5-phenoxy-10,15,20 triphenylporphyrin))), (CoPc-(CoTPP)4), containing four units of cobalt tetraphenylporphyrin linked to a central cobalt phthalocyanine macrocycle via ether linkages is reported. Cyclic voltammetry and spectroelectrochemistry were employed to characterize the complexes. Cyclic voltammetry and square wave voltammetry revealed nine processes. Spectroelectrochemistry of the pentamer confirmed that reduction occurs on the individual components in an alternating manner; the first reduction occurring on the CoTPP moieties, the second on the CoPc moiety, the third on the CoTPP moieties and so on. Oxidation occurred first on the CoPc moiety of the pentamer. PMID- 15782261 TI - Platinum complexes of diazo ligands. Studies of regioselective aromatic ring amination, oxidative halogen addition and reductive halogen elimination reactions. AB - 2-(Arylazo)pyridine ligands, L1a-1c react with the salt K2[PtCl4] to give the mononuclear complexes [PtCl2(L1)](1), which readily react with ArNH2 to yield the monochloro complexes of type [PtCl(L2)](HL2= 2-[(2 (arylamino)phenyl)azo]pyridine)(2) via regioselective ortho-amine fusion at the pendent aryl ring of coordinated L1. Oxidative addition of the electrophiles Y2(Y = Cl, Br, I) to the square-planar platinum(II) complex, has led to syntheses of the corresponding octahedral platinum(IV) complexes, [PtY3(L2)](3) in high yields. Ascorbate ion reductions of the platinum(IV) complexes, , resulted in reductive halogen elimination to revert to the platinum(II) complexes almost quantitatively. Isolation of products and X-ray structure determination of the representative complexes followed all these chemical reactions. In crystal packing, the compound [PtCl2(L1c)](1c) forms dimeric units with a Pt...Pt distance of 3.699(1) A. In contrast, the crystal packing of 2b revealed that the molecules are arranged in an antiparallel fashion to form a noncovalent 1D chain to accommodate pi(aryl)-pi(pyridyl) and Pt-pi(aryl) interactions. Notably, the oxidation of [Pt(II)Cl(L2a)](2a) by I2 produced a mixed halide complex [Pt(IV)ClI2(L2a)](5), which, in turn, is reduced by ascorbate ion to produce [Pt(II)I(L2a)] with the elimination of ClI. All the platinum(II) complexes are brown, the platinum(IV) complexes, on the other hand, are green. Low-energy visible range transitions in the complexes of the extended ligand [L2]- are ascribed to ligand basedpi-pi* transitions. Cyclic voltammetric behaviour of the complexes is reported. PMID- 15782262 TI - Trivalent lanthanide lacunary phosphomolybdate complexes: a structural and spectroscopic study across the series [Ln(PMo11O39)2]11-. AB - We report the syntheses and crystal structures of (NH4)11[Ln(III)(PMo11O39)2.xH2O (where Ln = every trivalent lanthanide cation except promethium) in which two lacunary [PMo11O39]7- anions sandwich an 8-coordinate Ln(III) cation to yield the complex anion, [LnIII(PMo11O39)2]11-. The 14 salts crystallise in two different space groups, C2/c or P1, but the LnIII containing anions are isostructural across the whole series, a very rare example of such a complete study. Solid state and solution 31P NMR, Raman and IR spectroscopies have been used to prove the stability of [Ln(PMo11O39)2]11- in aqueous solution. As expected, the LnIII cation contracts across the series and the Ln-O bond distances decrease uniformly. Interestingly, the splitting in the nu(P-O) mode within the [PMo11O39]7- unit increases uniformly across the series, which we attribute to the stronger interaction with the smaller, higher charge density LnIII cation as the series is traversed. For the 31P NMR measurements a direct comparison of Lanthanide Induced (paramagnetic) Shift could be made with the analogous [P(W11O39)2]11- complexes. PMID- 15782263 TI - The distinct affinity of cyclopentadienyl ligands towards trivalent uranium over lanthanide ions. Evidence for cooperative ligation and back-bonding in the actinide complexes. AB - The mono and bis(cyclopentadienyl) compounds [M(C5H4Bu t)I2] and [M(C5H4Bu t)2I](M = U, La, Ce, Nd) were formed in thf by comproportionation reactions of [M(C5H4Bu t)3] and LnI3 or [UI3(L)4](L = thf or py) in the molar ratio of 1 : 2 and 2 : 1, respectively, while treatment of [UI(3)(py)(4)] or LnI(3)(Ln = La, Ce, Nd) with 1 or 2 mol equivalents of LiC5H4Bu t in thf afforded the [M(C5H4Bu t)I2] and [M(C5H4Bu t)2I2]- compounds, respectively. The X-ray crystal structures of [M(C5H4Bu t)I2(py)3](M = U, La, Ce, Nd), [{Ce(C5H4Bu t)2(mu-I)}2] and [M(C5H4Bu t)2I(py)2](M = U, Nd) have been determined; the differences between the average M C distances in the mono(cyclopentadienyl) complexes correspond to the variation in the ionic radii of the trivalent uranium and lanthanide ions while the U-N and U-I bond lengths seem to be smaller than those predicted from a purely ionic bonding model. The distinct affinity of the cyclopentadienyl ligands towards Ln(III) and U(III) was revealed by two series of competing reactions: the ligand exchange reactions between [Ln(C5H4Bu t)(n')I(3-n')](Ln = La, Ce, Nd) and [U(C5H4Bu t)(n'')I(3-n'')] species (1 < or = n'+n'' =n < or = 5), and the addition of n mol equivalents of LiC(5)H(4)Bu(t)(1 [less-than-or-equal]n[less than-or-equal] 5) to a 1 : 1 mixture of LnI3 and [UI3(thf)4] or [UI3(py)4]. The stability of the [M(C5H4Bu t)I2] species was found to vary in the order Nd > Ce > U > La, a trend which is in accord with an electrostatic bonding model. However, the bis and tris(cyclopentadienyl) complexes of uranium are more stable than their lanthanide analogues. This difference can be accounted for by a higher degree of covalency in the U-C5H4Bu t bond, resulting from the late appearance of back-bonding which would emerge only after the first cyclopentadienyl ligand is bound. PMID- 15782264 TI - Structural studies of apatite-type oxide ion conductors doped with cobalt. AB - A series of Co doped lanthanum silicate apatite-type phases, La9.83Si4.5Co1.5O26, La9.66Si5CoO26, La10Si5CoO26.5 and La8BaCoSi6O26, have been synthesised, and neutron diffraction, EXAFS and XANES used to investigate their structures in detail. All compositions were shown to possess the hexagonal apatite structure, and the results confirmed that cobalt can be doped onto both the La and Si sites within the structure depending on the starting composition. The Co doping is shown to cause considerable local distortions within the apatite structure. In the case of Si site doping two compositions showed anisotropic peak broadening, which has been attributed to incommensurate ordering of oxygen within the apatite channels. PMID- 15782265 TI - EXAFS and time-resolved laser fluorescence spectroscopy (TRLFS) investigations of the structure of Cm(III)/Eu(III) complexed with di(chlorophenyl)dithiophosphinic acid and different synergistic agents. AB - The complexes of trivalent actinide curium (Cm(III)) with di(chlorophenyl)dithiophosphinic acid ((ClPh)2PSSH) and three different neutral complexing agents as synergists in tert-butylbenzene are studied by EXAFS and time-resolved laser fluorescence spectroscopy (TRLFS). The results are compared with those from the corresponding europium (Eu(III)) complexes. The aim of these investigations is to understand the chemical interactions responsible for the high selectivity of the synergistic systems of (ClPh)2PSSH and neutral complexing agents tri-n-octylphosphine oxide, tributylphosphate and tris(2 ethylhexyl)phosphate for trivalent actinide cations in liquid-liquid extraction. In our structural chemistry study, we find that the inner coordination sphere of extracted Cm(III) and Eu(III) complexes are different. In all complexes the (ClPh)2PSSH is bound to the metal cation in a bidentate fashion and the oxygen donor of the neutral complexing agent used as synergist is directly coordinated to the metal cation. Comparison of the Cm(III) and Eu(III) complexes shows that Cm(III) preferentially binds to the sulfur of (ClPh)2PSSH, whereas Eu(III) is preferentially bound to oxygen. A good selectivity in liquid-liquid extraction is correlated with a high ratio of the sulfur coordination number to oxygen coordination number. This leads to the conclusion that the observed differences in the coordination structure between Cm(III) and Eu(III) complexes play an important role in the selectivity of these extraction systems. PMID- 15782266 TI - Synthesis and characterisation of tungsten(VI) oxo-salicylate complexes for use in the chemical vapour deposition of self-cleaning films. AB - Tungsten(VI) oxo-salicylate complexes were prepared in moderate yield (47 to 63%) by the reactions of WOCl4 and two equivalents of either 3-methylsalicylic acid (MesaliH2) or 3,5-di-isopropylsalicylic acid (di-i-PrsaliH2). Performing the reaction in refluxing toluene afforded the two analogous ditungsten complexes 1, [{WO(Mesali)(MesaliH)}2(mu-O)], and 2, [{WO(di-i-Prsali)(di-i-PrsaliH)}2(mu-O)], however in refluxing hexane the mononuclear tungsten complex , [WO(di-i Prsali)(di-i-PrsaliH)Cl], was isolated. The single crystal X-ray study of revealed a pseudo-octahedral geometry around the tungsten centres. Aerosol assisted chemical vapour deposition of or afforded brown tungsten trioxide thin films. These films were converted to yellow fully oxidised WO3 on annealing in air at 550 degrees C for 30 minutes. The yellow WO3 films demonstrate preferred orientation on the substrate and show interesting functional properties-photo induced hydrophilicity and photocatalytic activity. PMID- 15782267 TI - The electron-poor phosphines P{C6H3(CF3)2-3,5}3 and P(C6F5)3 do not mimic phosphites as ligands for hydroformylation. A comparison of the coordination chemistry of P{C6H3(CF3)2-3,5}3 and P(C6F5)3 and the unexpectedly low hydroformylation activity of their rhodium complexes. AB - The fluoroaryl phosphines P{C6H3(CF3)2-3,5}3 (La) and P(C6F5)3 (Lb) form the complexes trans-[MCl2(La)2] and trans-[MCl2(Lb)2](M = Pd or Pt) which have been isolated and fully characterised. 31P NMR studies of competition experiments show that the stability of trans-[PdCl2L2] is in the order L = Lb< La /=5-mm increase in such distance was considered indicative of device migration. Migration cumulative incidence was estimated by the Kaplan-Meier method, and its association with any of the preoperative anatomical characteristics was tested using Cox proportional hazards models. Median follow-up time was 9 (range, 1-31) months. Migration occurred in nine patients, corresponding to a 15.6% estimated probability of migration at 30 months (SE = 5.1%). Migration was associated with the risk of proximal type I endoleak (hazard ratio = 3.39, 95% confidence interval = 1.46-7.87; p = 0.007). This type of endoleak occurred in three of the migration-affected patients (33.3%); all of them were resolved by additional cuff placement at the proximal landing zone. No other migration-related reinterventions were performed. The only significant associations between anatomic factors and device migration probability were the protective effects of longer necks (odds ratio [OR] = 0.71 for each additional 5 mm, p = 0.045) and longer overlapped portions of neck and device (OR = 0.56 for each additional 5 mm, p = 0.003). There was a trend toward higher probability of migration among reverse-tapered necks (OR = 1.75, p = 0.109). Percentage of device oversizing correlated with early neck dilation (between preoperative and first postoperative diameters, correlation coefficient = 0.4, p < 0.0001), but not with late neck dilatation (between first postoperative and 1.5-year scan diameters, correlation coefficient = 0.29, p = 0.112). There was a trend toward higher mean percentage of late dilation among migrators (11.4%, standard error of the mean [SEM] 2.6) than nonmigrators (5.7%, SEM = 1) (p = 0.08), but both groups had similar mean percentages of early dilation (3%, SEM = 1.6%, vs. 5.5%, SEM = 0.6%; p = 0.365). This result indicates that device migration is not a rare event after AneuRx implantation. This phenomenon is associated with proximal type I endoleaks. Deployment of the endograft immediately below the renal arteries might help to prevent migration, since use of greater lengths of overlapped device relative to the proximal neck has a protective effect. Migration seems to be independent of the degree of device oversizing. PMID- 15782272 TI - Comparison of bedside transabdominal duplex ultrasound versus contrast venography for inferior vena cava filter placement: what is the best imaging modality? AB - While contrast venography is considered the gold standard for imaging prior to inferior vena cava (IVC) filter insertion, bedside placement via transabdominal duplex ultrasound (DUS) has been recognized as a safe and effective alternative. To date, there has been no direct comparison of the efficacy of both imaging modalities for IVC filter placement. A concurrent cohort of patients who underwent IVC filter placement at a single institution over a 7-year period with either contrast venography or transabdominal DUS performed at bedside was retrospectively reviewed. Patient demographics, venous thromboembolism risk factors, indications, technical success, and procedural complications were compared. Of 439 patients initially imaged with transabdominal DUS, IVC filter placement was determined to be technically feasible in 382 patients (87%). The procedural technical success rate for IVC filter placement using transabdominal DUS when IVC visualization was adequate was 97.4% (n = 382 patients), compared to 99.7% (n = 318 patients) for contrast venography (p = 0.018). Patients undergoing IVC filter placement with transabdominal DUS more commonly required IVC filter for venous thromboembolism prophylaxis (81.1% vs. 27.8%, p < 0.001), had increased incidence of multiple traumatic injuries (28% vs. 10%, p < 0.001), and had increased risk from immobilization (91.3% vs. 34.1%, p < 0.001). Overall complication rates were 0.6% for venography and 1.8% for transabdominal DUS (p = NS). When IVC visualization was adequate, contrast venography and transabdominal duplex ultrasound both had high rates of success and a low incidence of complications. A technical success advantage was observed for contrast venography; this difference in technical success must be weighed against the bedside insertion advantage offered by DUS, which may be especially important in the immobilized or critically ill patient. Transabdominal DUS remains our preferred technique when feasible, especially when bedside placement is desired. PMID- 15782273 TI - Risk-adjusted analysis of outcomes following elective open abdominal aortic aneurysm repair. AB - The purpose of this study was to describe a method to analyze outcomes following open abdominal aortic aneurysm (AAA) repair while considering the variability in patients' preoperative risk. Consecutive patients undergoing elective open infrarenal AAA repair during a 4-year period (2000-2003) were reviewed. Thirty day or in-hospital mortality was the major outcome variable. Preoperative mortality risk was estimated for each patient using a validated scoring system that considers age, renal dysfunction, and coronary artery and cerebrovascular disease. A risk-adjusted cumulative sum method was used to compare observed versus predicted outcomes by assigning a risk-adjusted score, based on log likelihood ratios, to each patient. These cumulative scores were sequentially plotted with preset control limits to allow for "signaling" when results were substantially different than expected (doubling or halving of odds ratios). Four hundred and sixty-three patients were studied with an overall early mortality rate of 4.5% (n = 21). Patients were allocated to three different preoperative risk groups (low, n = 89; medium, n = 160; high, n = 214) according to a medical comorbidity-based scoring system. Predicted (P) and observed (O) mortality rates for each group were as follows: low, 2.4% (P) and 2.2% (O); medium, 4.1% (P) and 4.4% (O); high, 9.3% (P) and 5.6% (O). The resulting risk-adjusted scores for each patient were plotted sequentially. This plot was flat for the first year and then adopted a negative slope crossing the lower control limit after 266 patients, indicating improved results compared to those expected. This coincided with the adoption of routine intraoperative cell saver use in our practice. This form of analysis allows for the prospective evaluation of results while considering patient-mix variabilities. PMID- 15782274 TI - Percutaneous endovascular treatment for chronic limb ischemia. AB - Percutaneous transluminal methods of revascularization for chronic limb ischemia have traditionally been used as an adjunctive or secondary mode of treatment. This study examines the outcomes of patients treated via endovascular methods as the primary form of treatment. The records of 66 patients (average age, 71.3 years; range, 53 to 92) with chronic limb ischemia treated from November 2000 to January 2002 at a single institution with endovascular methods were retrospectively reviewed. Thirty-one patients had disabling claudication, 15 had rest pain, 17 had minor tissue loss, and 3 had major tissue loss. There were 94 lesions in the 66 limbs: 45 lesions in the superficial femoral artery, 28 lesions in the popliteal artery, and 21 lesions in tibial arteries. Occlusive lesions were treated by subintimal angioplasty. Stenoses were treated by percutaneous transluminal angioplasty (PTA). The median length of the lesions was 12 cm. Forty six stents were placed in 32 patients; 34 patients did not receive stents. The approach was contralateral in 71% (47/66) and ipsilateral in 29% (19/66) of cases. Patients were followed up with physical exam, anklebrachial index (ABI), and duplex ultrasonography. Post-intervention medical treatment included aspirin and clopidogrel. Long-term patency was correlated with age, comorbidities, risk factors, and the use of stents in addition to angioplasty. Endovascular treatment was technically successful in 65 of 66 patients (98%). Residual stenosis (<30%) was found in three cases, two in patients with PTA alone and one patient with PTA and a stent. Mean length of follow-up was 6.3 months (range, 6 weeks to 15 months). Post-intervention ABI improved from a mean of 0.65 to a mean of 1.01 (p < 30.05). The perioperative complication rate was 9% and included three cases of hematoma and one case each of thrombosis, lymphocele, and rash reaction to medications. Outcome measures included primary patency (89%), assisted primary patency (96%), and limb salvage (97%) at 6 months. Restenosis of >50% occurred in four patients and occlusion occurred in two. At 6 months, patients with one or more stents had a patency rate of 84% (27/32), whereas those without a stent had a patency rate of 94% (32/34) (p = NS). Endovascular treatment is a safe, feasible, and effective method with a high short-term patency rate and may be used as an alternative to bypass for chronic limb ischemia. PMID- 15782275 TI - Spontaneous pseudoaneurysm of the vertebral artery in Behcet's disease. AB - A pulsatile mass and severe neck pain developed suddenly in a 15-year-old female patient suffering from Behcet's disease. Magnetic resonance imaging showed a pseudoaneurysm at the C3-C4 level that was 51 x 49 x 45 mm in size, originating from the left vertebral artery, with a thin neck and thrombus inside. Repair of the vertebral artery wall by percutaneous transluminal intervention was not successful. Because of the possibility of rupture, the patient underwent surgical repair. Ligation of the left vertebral artery was applied 1 cm above the origin of the subclavian artery. During the subsequent postoperative period, no further complications were seen. From our review of the literature, this is the first reported case of surgical treatment of spontaneous development of a pseudoaneurysm at the vertebral artery in association with Behcet's disease. Ligation of the vertebral artery can be safely used to control a pseudoaneurysm related to Behcet's disease. PMID- 15782276 TI - Frequency of Norovirus in stool samples from patients with gastrointestinal symptoms in Switzerland. AB - To determine the frequency of sporadic community-acquired Norovirus (NV) infection in the German-speaking part of Switzerland, an evaluation of gastroenteritis cases seen in physicians' practices between July 2001 and July 2003 was conducted. A total of 699 stool specimens documented to be free of common bacterial pathogens was screened for the presence of NV by RT-PCR. NV was detected in 125 (17.9%) of these specimens. In the seasonal analysis, the highest rate of NV-positive samples (38.3%) was found between January and March 2002. After July 2002, the study was expanded to additionally test for NV in stool samples containing a known bacterial pathogen. Among 132 such specimens, NV was detected in only one. This suggests that NV mixed-infections are playing a marginal role in Switzerland. PMID- 15782277 TI - Prophylactic use of antibiotics for prevention of meningococcal infections: systematic review and meta-analysis of randomised trials. AB - A systematic review and meta-analysis of randomised controlled trials was performed in order to study the effectiveness of prophylactic treatment regimens in preventing secondary cases of meningococcal disease (i.e., those contracted after contact with a person with meningococcal disease) and in eradicating Neisseria meningitidis from the pharynx of healthy colonised individuals. The Medline, Embase, and Lilacs databases, the Cochrane Library, and the references of all studies identified were systematically searched for relevant trials. Two reviewers independently applied selection criteria, performed quality assessment, and selected data. Relative risks were pooled using a fixed effects model unless heterogeneity assessed by the I2 statistic and chi-square test was found. In such cases, a random effect model was used. There were no cases of meningococcal disease following treatment with antibiotics or placebo, thus effectiveness regarding prevention of future disease could not be directly assessed. Compared with placebo, ciprofloxacin (RR = 0.04; 95%CI, 0.01-0.12), rifampin (RR = 0.17; 95%CI, 0.12-0.24), minocycline (RR = 0.30; 95%CI, 0.19-0.45), and penicillin (RR = 0.47; 95%CI, 0.24-0.94), proved effective at eradicating Neisseria meningitidis 1 week after treatment. After 1-2 weeks, only ciprofloxacin (RR = 0.03; 95%CI, 0.00-0.42) and rifampin (RR = 0.20; 95%CI, 0.14-0.29) still proved significantly effective when compared with placebo. Rifampin continued to be effective (RR = 0.24; 95%CI, 0.16-0.37) compared with placebo until up to 4 weeks post treatment. Rifampin was the only drug to which resistance developed. Given that the use of rifampin in an outbreak setting might lead to the circulation of isolates resistant to rifampin, the use of ciprofloxacin and ceftriaxone should be considered. PMID- 15782278 TI - Lower mortality among patients with community-acquired pneumonia treated with a macrolide plus a beta-lactam agent versus a beta-lactam agent alone. AB - A cohort of 1,391 patients with community-acquired pneumonia of unknown etiology, atypical pneumonia, Legionella pneumophila pneumonia, viral pneumonia, or pneumococcal pneumonia was studied according to a standard protocol to analyse whether the addition of a macrolide to beta-lactam empirical treatment decreases mortality rates. Patients admitted to the intensive care unit were excluded. Severity was assessed using the PORT score. An etiological diagnosis was achieved in 498 (35.8%) patients (292 infections due to Streptococcus pneumoniae). Treatment was chosen by the attending physician according to his/her own criteria: beta-lactam agent in 270 and beta-lactam agent plus a macrolide in 918 cases. The mortality rate was 13.3% in the group treated with a beta-lactam agent alone and 6.9% in the group treated with a beta-lactam agent plus a macrolide (p=0.001). The percentage of PORT-group V patients was 32.6% in the group treated with a beta-lactam agent alone compared to 25.7% in the group who received a beta lactam agent plus a macrolide (p=0.02). After controlling for PORT score, the odds of fatal outcome was two times higher in patients treated with a beta-lactam agent alone than in those treated with a beta-lactam agent plus a macrolide (adjusted OR = 2, 95%CI 1.24-3.23). The results suggest that the addition of a macrolide to an initial beta-lactam-based antibiotic regimen is associated with lower mortality in patients with community-acquired pneumonia, independent of severity of infection, thus supporting the recommendation of a beta-lactam-agent plus a macrolide as empirical therapy. PMID- 15782279 TI - Chlamydophila pneumoniae antibodies in office workers with and without inflammatory rheumatic diseases in a moisture-damaged building. PMID- 15782280 TI - Congenital Spigelian hernia with cryptorchidism: probably a new syndrome. AB - Nearly 28% of pediatric Spigelian hernias reported in the literature are associated with ipsilateral cryptorchidism. However, the pathogenetic relationship between the two has not been satisfactorily explained in the past. This paper describes a male neonate born with cryptorchidism and imperforate anus. Anal stenosis following the treatment of imperforate anus had let to the development of multiple hernias including Spigelian hernia on the right side. Surgical exploration revealed the right testis being located within the Spigelian hernia. Based on the sequence of events, it is hypothesized that Spigelian hernia in this case is a sequela of maldescended testis and raised intraabdominal pressure. As this explanation is also applicable to all of the previously reported cases, the author suggests that the combination of Spigelian hernia and ipsilateral cryptorchidism could probably form a hitherto unrecognized new syndrome. PMID- 15782281 TI - Axial BMD, change in BMD and bone turnover do not predict breast cancer incidence in early postmenopausal women. AB - Previous studies have indicated a relationship between bone mineral density and the incidence of breast cancer in middle-aged and elderly women, with women with higher BMD being at significant increased risk. We investigated whether there was such a relationship in younger women who were perimenopausal or in their early postmenopausal years. As part of a population-screening program for osteoporosis, 5,119 women aged between 45 and 54 years were scanned between 1990-1994 at the Osteoporosis Research Unit. In 1997-2001, 3,884 returned for follow-up scans and questionnaires, and 3,144 returned a postal questionnaire in 2002. All cases of incident breast cancer were noted. One hundred sixty-six women indicated that they had suffered from breast cancer, of which 87 were incident cases (59 had prevalent breast cancer at baseline and 20 had benign or unconfirmed diagnosis and were excluded because of the use of agents that may interfere with BMD, e.g., tamoxifen). We compared therefore the incident breast cancer group (BC group; n=87) with a control group (C group; n=3,013). There were no significant differences using a t-test between the BC group and C group for baseline DXA of the spine or femoral neck. Further changes in BMD over a mean period of 6.9 years demonstrated no significant hazard ratio for the lumbar spine or femoral neck. No relationship was seen between the bone turnover markers pyridinoline/creatinine or deoxypyridinoline/creatinine assessed at their second study visit and incidence of breast cancer. In conclusion, in perimenopausal or early postmenopausal women there is no relationship between the incidence of breast cancer and BMD, change in BMD or bone turnover. PMID- 15782282 TI - The influence of Lys3Asn polymorphism in the osteoprotegerin gene on bone mineral density in Chinese postmenopausal women. AB - The objective was to identify single nucleotide polymorphisms (SNPs) in exons of the osteoprotegerin gene and to analyze the relationship between the SNPs and bone mineral density in postmenopausal women. We used polymerase chain reaction (PCR) and direct sequencing methods to identify SNPs and genotypes in 205 postmenopausal women. BMD at the lumbar spine (L2-4) and femoral neck (FN) were measured by dual-energy X-ray absorptiometry (DEXA). Serum osteocalcin (OC), osteoprotegerin (OPG), receptor activator of nuclear factor kappaB ligand (RANKL) and urinary N-telopeptide of type I collagen (NTx) were also measured. In exon 1 of the OPG gene, we found the Lys3Asn SNP. In 205 postmenopausal women, the Asn allele frequency was 26.0%, and the distribution of Lys3Asn genotypes was Lys-Lys 56.6%, Lys-Asn 34.6% and Asn-Asn 8.8%, respectively. BMD at the lumbar spine (L2 4) of the Asn-Asn genotype was significantly higher (9.5-12.6%) than Lys-Asn and Lys-Lys genotypes (P=0.012), with evidence for an allele dose effect (P=0.008). Results remained similar after adjustment for age and body mass index. The Lys3Asn polymorphism of the OPG gene alone accounted for 7.7% of the variance of the L2-4 BMD in a multiple regression model. Logistic regression analysis revealed that the OPG genotype Lys-Lys had a 2.7 times (95% CI: 0.83-9.11) greater risk for osteopenia/osteoporosis than the Asn-Asn genotype. The Lys3Asn polymorphism in the OPG gene is associated with L2-4 BMD in postmenopausal women. The Lys-allele is associated with lower BMD and an increased risk for osteoporosis. PMID- 15782283 TI - Body size from birth to adulthood and bone mineral content and density at 31 years of age: results from the northern Finland 1966 birth cohort study. AB - The purpose of this study was to evaluate the association between body size from birth to adulthood and bone mineral content (BMC) and bone mineral density (BMD) at the age of 31 years in a longitudinal study of the Northern Finland birth cohort for 1966. Data were collected at birth, 1, 14, and 31 years. This analysis was restricted to a subsample of individuals (n =1,099) for whom the BMC (g) and BMD measurements (g/cm(2)) were performed on the distal and ultradistal radius by dual-energy X-ray absorptiometry (DXA) at the age of 31 years. Determinants of low BMC and BMD were analyzed using multivariate logistic regression. Growth retardation at birth, being underweight (BMI < or =20.0 kg/m(2)) at 31 years, and having a low calcium intake at 31 years were associated independently with low BMD at 31 years. Additionally, the proportion of subjects with low BMD was higher among those who had low standardized body weight (< or =1 SD) both at birth and at 14 years, and both at 14 and 31 years. Body weight at 31 years was the strongest associating factor of BCM at 31 years. Growth retardation at birth has long-lasting effects on adult bone mineral content and density of the distal and ultradistal radius independently of later body size, although adult body weight seems to be a most important determinant of BMC at the age of 31 years. Thinness and a low calcium intake are associated with low bone mineral content and density at 31 years of age. Further studies are needed to evaluate if these groups are at increased risk of osteoporosis in old age. PMID- 15782285 TI - Comparison of spine and femur reference data in native Chinese women from different regions of China. AB - The aim of this study is to explore the differences of BMD reference curves at various skeletal sites among Chinese women from different regions of China and to investigate the feasibility of establishing a unified national BMD reference database for Chinese women. We measured BMD at the posteroanterior (PA) lumbar spine, femoral neck, trochanter and Ward's triangle by dual-energy X-ray absorptiometry bone densitometer in 3,422 Changsha women of South Central China, aged 20-84 years. The documented BMDs of reference populations of women in all other areas included Shanghai ( n =2,111) and Nanjing ( n =3,174) in the East, Shenyang ( n =1,213) in the Northeast, Kunming ( n =523) in the Southwest, Chongqing ( n =811) in the Midwest and Xian ( n =1,320) in the Northwest. We adopted the cubic regression as the fitting model for reference curves of BMD that varied with age, conducted conversions of BMD measured by various bone densitometers from different manufacturers and compared the differences between standardized BMD (sBMD) reference curves and combined ones for women from different areas. Our results revealed that by comparing variances in women from different areas, the average variances of non-standard BMD were 0.8-30.8% at the PA spine, 0.7-24.5% at the femoral neck, 0.6-29.9% at the trochanter and 1.1 54.7% at Ward's triangle, while average variances of sBMD either significantly decreased or disappeared (0.8-3.9% at the PA spine, 0.7-8.6% at the femoral neck, 0.6-8.3% at the trochanter and 1.1-29.9% at Ward's triangle). The sBMD reference curves were highly positive-dependent with combined ones ( r =0.913-0.999, P =0.000). At the PA spine and trochanter, the effect of combined sBMD curves presented well in women from different areas, except for those from Shanghai at the PA spine and Shenyang at the trochanter, indicating that sBMD curves were significantly different from pooled ones; at the femoral neck and Ward's triangle, the effect of combined sBMD reference curves was poor, indicating that sBMD curves demonstrated significant differences from pooled ones in women from a majority of these areas. We conclude that, in high density population areas, sBMD reference curves showed no significant geographic differences in women from various regions. In women from different areas, sBMD reference curves present good pooled results at the PA spine and trochanter. The less ideal combining effect of the sBMD curves at both femoral neck and Ward's triangle might be caused by the intrinsic differences from the different measuring instruments. PMID- 15782284 TI - Diet and exercise during growth have site-specific skeletal effects: a co-twin control study. AB - Exercise and improved nutrition offer safe, low-cost and widely applicable approaches to potentially reduce the burden of fractures. We conducted a cross sectional study of 30 monozygotic and 26 dizygotic male twin pairs, aged 7-20 years to test the following hypotheses: (1) Associations between bone mass and dimensions and exercise are greater than between bone mass and dimensions and protein or calcium intakes; (2) exercise or nutrient intake are associated with appendicular bone mass before puberty and axial bone mass during and after puberty. Total body and posteroanterior (PA) lumbar spine bone mineral content (BMC) and mid-femoral shaft dimensions were measured using dual energy X-ray absorptometry (DEXA). Relationships between within-pair differences in nutrient intake (determined by weighed-food diaries) or exercise duration (determined by questionnaire) and within-pair differences in BMC and bone dimensions were tested using linear regression analysis. In multivariate analyses, within-pair differences in exercise duration were associated with within-pair differences in total body, leg and spine BMC, and cortical thickness. Every-hour-per-week difference in exercise was associated with a 31-g (1.2%) difference in total body BMC, a 10-g (1.4%) difference in leg BMC, a 0.5-g difference in spine BMC and a 0.1-mm difference in cortical thickness ( p <0.01- p <0.1). A 1-g difference in protein intake was associated with a 0.8-g (0.4%) difference in arm BMC ( p <0.05). These relationships were present in peri-pubertal and post-pubertal pairs but not in pre-pubertal pairs. Exercise during growth appears to have greater skeletal benefits than variations in protein or calcium intakes, with the site specific effects evident in more mature twins. PMID- 15782286 TI - Assessment of pelvic floor movement using transabdominal and transperineal ultrasound. AB - The aims of the study were (1) to assess the reliability of transabdominal (TA) and transperineal (TP) ultrasound during a pelvic floor muscle (PFM) contraction and Valsalva manoeuvre and (2) to compare TA ultrasound with TP ultrasound for predicting the direction and magnitude of bladder neck movement in a mixed subject population. A qualified sonographer assessed 120 women using both TA and TP ultrasound. Ten women were tested on two occasions for reliability. The reliability during PFM was excellent for both methods. TP ultrasound was more reliable than TA ultrasound during Valsalva. The percentage agreement between TA and TP ultrasound for assessing the direction of movement was 85% during PFM contraction, 100% during Valsalva. There were significant correlations between the magnitude of the measurements taken using TA and TP ultrasound and significant correlations with PFM strength assessed by digital palpation. PMID- 15782287 TI - Influence of quercetin and rutin on growth and antioxidant defense system of a human hepatoma cell line (HepG2). AB - BACKGROUND: Dietary polyphenols like quercetin and rutin are considered beneficial because of their potential protective role in the pathogenesis of multiple diseases associated to oxidative stress such as cancer, coronary heart disease and atherosclerosis. However, many of these effects may depend on the concentration of the polyphenol utilized since high doses of some phenolic compounds may be prooxidant and negatively affect cell growth and viability. AIM OF THE STUDY: To test the potential chemoprotective effects of quercetin and rutin, two flavonols with high antioxidant capacity, on cell growth, viability and the response of the antioxidant defense system of a human hepatoma cell line (HepG2). METHODS: Cell growth was measured by diaminobenzoic acid and bromodeoxyuridine assays, cell toxicity by lactate dehydrogenase leakage assay, reduced glutathione was quantified by a fluorimetric assay, cellular malondialdehyde was analyzed by high-performance liquid chromatography, reactive oxygen species were quantified by the dichlorofluorescein assay, antioxidant enzyme activities were determined by spectrophotometric analysis and their gene expression by northern blot. RESULTS: Short-term exposure (4 h) to these flavonols had no antiproliferative nor cytotoxic effect. High doses of quercetin (50-100 microM) increased glutathione concentration and gene expression of Cu/Zn superoxide dismutase and catalase inhibiting the activity of the latter enzyme, whereas lower doses (0.1-1 microM) decreased gene expression of Cu/Zn superoxide dismutase and increased that of glutathione peroxidase. All doses of quercetin and rutin diminished reactive oxygen species and high doses (10-100 microM) decreased malondialdehyde concentration. CONCLUSION: The results indicate that both natural antioxidants induce favorable changes in the antioxidant defense system of cultured HepG2 that prevent or delay conditions which favor cellular oxidative stress. PMID- 15782288 TI - Rosa roxburghii supplementation in a controlled feeding study increases plasma antioxidant capacity and glutathione redox state. AB - BACKGROUND: Rosa roxburghii (RR) is a plant of which the fruit juice has been used as a medicinal remedy for a variety of diseases. It has been proposed that the putative beneficial properties are related to its antioxidant potential. AIM OF STUDY: We investigated the contribution of a supplemented RR fruit sample on the antioxidant status in a cohort of healthy humans. METHODS: A total of 36 young, healthy and non-smoking individuals were recruited for this randomised placebo-controlled, single-blind trial. The study was diet controlled over a five week period with a two week run-in period before participants daily received a placebo or an encapsulated supplement of RR sample. Total antioxidant capacity, glutathione redox state, glutathione reductase, glutathione peroxidase, superoxide dismutase and 8-OHdG levels were measured. RESULTS: RR supplementation significantly increased plasma antioxidant capacity (p = 0.04) and GSH:GSSG ratios in blood (p = 0.03). No significant changes in 8-OHdG levels, total glutathione levels or antioxidant modulating enzymes were detected suggesting that the observed shift of the glutathione redox state probably occurs via the antioxidant mediated protection of GSH. CONCLUSIONS: We conclude that these findings support the putative beneficial properties that have been linked to Rosa roxburghii as a dietary supplement that can enhance antioxidant status. PMID- 15782289 TI - Low temperature stimulates cell enlargement and intracellular calcification of coccolithophorids. AB - Temperature effect on growth, cell size, calcium uptake activity, coccolith production was studied in coccolith-producing haptophytes, Emiliania huxleyi (Lohmann) Hay & Mohler (strain EH2) and Gephyrocapsa oceanica Kamptner (strain GO1) (Coccolithophorales, Prymnesiophyceae). E. huxleyi grew at a wider temperature range (10 degrees-25 degrees C), while G. oceanica growth was limited to warmer temperatures (20 degrees-25 degrees C). Cell size was inversely correlated with temperature. At low temperature, the enlargement of chloroplasts and cells and the stimulation of coccolith production were morphologically confirmed under fluorescent and polarization microscopes, respectively. 45Ca uptake by E. huxleyi at 10 degrees C was greatly increased after a 5-day lag and exceeded that at 20 degrees C. These results clearly showed that low temperature suppressed coccolithophorid growth but induced cell enlargement and as stimulated the intracellular calcification that produces coccoliths. PMID- 15782290 TI - [Diseases of the lacrimal gland]. AB - Experience with the differential diagnosis of lacrimal gland illnesses is limited in daily practice due to their rarity. It is therefore of particular importance to have guidelines for the practitioner by which he can decide on the application of conservative or surgical therapy, and whether it is necessary to take a biopsy to clarify the diagnosis. There is a wide spectrum of possible illnesses which include systemic diseases, inflammatory disorders, neoplastic, benign and malignant tumors. When a pleomorphic adenoma cannot be excluded, biopsy is contraindicated and complete tumor excision with its capsule is necessary to prevent the possibility of malignant transformation and a negative effect on long term prognosis. In this overview, special interest is placed on the typical clinical aspects and imaging features of lesions of the lacrimal gland fossa, their differential diagnosis and management. PMID- 15782291 TI - [Genetics of retinal dystrophies--an overview]. AB - Vision requires complex retinal functions, involving multiple genes with different functions. Retinal degeneration results from disturbance of retina specific processes such as the visual transduction cascade, but also from defects in basic functions such as pre-mRNA splicing and nucleotide synthesis. As a consequence, the retinal dystrophies are genetically extremely heterogeneous (as shown in the table). Thanks to the Human Genome Project, the identification of retinal disease genes and additional loci has skyrocketed. Today, a typical search for the causative gene in a disease-linked genomic interval starts at the computer. Genes from a particular region can be displayed, and multiple gene specific data such as expression patterns are immediately accessible. Candidate genes can then be investigated in DNA from affected individuals. PMID- 15782292 TI - Test systems to study transcriptional regulation and promoter activity in Bacillus megaterium. AB - Plasmid-located (multi-copy) and chromosomally located (single-copy) promoter test systems were developed for Bacillus megaterium by making use of the homologous beta-galactosidase-encoding bgaM gene. The multi-copy system facilitates rapid promoter analyses and promoter trapping, whereas the single copy system, integrated into the chromosome, allows investigation of tightly regulated promoters. As a prerequisite for both the multi- and the single-copy systems, a beta-galactosidase-deficient B. megaterium strain was generated by deletion mutagenesis. Both test systems were verified using the promoter of the xylose operon (P( xylA )) from B. megaterium along with its repressor (XylR). As expected, expression levels in the two systems differed significantly, although expression of the bgaM reporter gene was induced by xylose in both cases, thereby proving the functionality of both the multi- and the single-copy system. PMID- 15782293 TI - The importance of aeration strategy in fuel alcohol fermentations contaminated with Dekkera/Brettanomyces yeasts. AB - Whole corn mash fermentations infected with industrially-isolated Brettanomyces yeasts were not affected even when viable Brettanomyces yeasts out-numbered Saccharomyces yeasts tenfold at the onset of fermentation. Therefore, aeration, a parameter that is pivotal to the physiology of Dekkera/Brettanomyces yeasts, was investigated in mixed culture fermentations. Results suggest that aeration strategy plays a significant role in Dekkera/Brettanomyces-mediated inhibition of fuel alcohol fermentations. Although growth of Saccharomyces cerevisiae was not impeded, mixed culture fermentations aerated at rates of > or =20 ml air l(-1) mash min(-1) showed decreased ethanol yields and an accumulation of acetic acid. The importance of aeration was examined further in combination with organic acid(s). Growth of Saccharomyces occurred more rapidly than growth of Brettanomyces yeasts in all conditions. The combination of 0.075% (w/v) acetic acid and contamination with Brettanomyces TK 1404W did not negatively impact the final ethanol yield under fermentative conditions. Aeration, however, did prove to be detrimental to final ethanol yields. With the inclusion of aeration in the control condition (no organic acid stress) and in each fermentation containing organic acid(s), the final ethanol yields were decreased. It was therefore concluded that aeration strategy is the key parameter in regards to the negative effects observed in fuel alcohol fermentations infected with Dekkera/Brettanomyces yeasts. PMID- 15782294 TI - Arthritis in Kawasaki disease after responding to intravenous immunoglobulin treatment. PMID- 15782295 TI - Reduced-antigen-content-diphtheria-tetanus-acellular-pertussis and inactivated polio vaccine as a booster for adolescents 10 to 14 years of age. AB - High rates of pertussis disease in adolescents suggest that additional boosting against pertussis would be beneficial. A combined acellular-pertussis-containing booster vaccine (dTpa-IPV; Boostrixtrade mark Polio, n =440) was compared to separately administered dTpa (Boostrixtrade mark) and inactivated polio virus (IPV; Imovax Polio((R)), n =219), and to DTPa-IPV (Infanrixtrade mark IPV, n =111) vaccine in a partially blind, randomised controlled trial in 10-14 year olds. One month after vaccination, seroprotection/seropositivity rates for all antigens were similar for all groups. Although pertussis and diphtheria antibody geometric mean antibody concentrations were higher after DTPa-IPV, all subjects had protective antibodies against diphtheria, tetanus and polio, and at least 97% had a vaccine response to pertussis antigens. Reactogenicity of dTpa-IPV was comparable to dTpa + IPV, but dTpa-IPV was generally better tolerated than DTPa IPV. CONCLUSION: The combined reduced-antigen-content-diphtheria-tetanus acellular-pertussis and IPV vaccine is immunogenic and well tolerated when administered to adolescents and could be used to improve the control of pertussis disease in this age group. PMID- 15782296 TI - A first linkage map of pecan cultivars based on RAPD and AFLP markers. AB - We report here the first genetic linkage maps of pecan [Carya illinoinensis (Wangenh.) K. Koch], using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers. Independent maps were constructed for the cultivars 'Pawnee' and 'Elliot' using the double pseudo-testcross mapping strategy and 120 F1 seedlings from a full-sib family. A total of 477 markers, including 217 RAPD, 258 AFLP, and two morphological markers were used in linkage analysis. The 'Pawnee' linkage map has 218 markers, comprising 176 testcross and 42 intercross markers placed in 16 major and 13 minor (doublets and triplets) linkage groups. The 'Pawnee' linkage map covered 2,227 cM with an average map distance of 12.7 cM between adjacent markers. The 'Elliot' linkage map has 174 markers comprising 150 testcross and 22 intercross markers placed in 17 major and nine minor linkage groups. The 'Elliot' map covered 1,698 cM with an average map distance of 11.2 cM between adjacent markers. Segregation ratios for dichogamy type and stigma color were not significantly different from 1:1, suggesting that both traits are controlled by single loci with protogyny and green stigmas dominant to protandry and red stigmas. These loci were tightly linked (1.9 cM) and were placed in 'Elliot' linkage group 16. These linkage maps are an important first step towards the detection of genes controlling horticulturally important traits such as nut size, nut maturity date, kernel quality, and disease resistance. PMID- 15782297 TI - Genetic analysis of adaptive syndromes interrelated with seed dormancy in weedy rice (Oryza sativa). AB - Seed dormancy in rice interrelates to the weedy characteristics shattering, awn, black hull color, and red pericarp color. A cross between the weedy strain SS18-2 and the breeding line EM93-1 was developed to investigate the genetic basis and adaptive significance of these interrelationships. These characteristics or their components differed in average degree of dominance from -0.8 to 1.5, in heritability from 0.5 to 0.96, and in their contribution to phenotypic or genotypic variation in dormancy by up to 25%. Five dormancy, four shattering, and three awn-length quantitative trait loci (QTLs) were detected in the BC1 population replicated in 2 years. Two QTLs for hull color were identified, and the SS18-2-derived and EM93-1-derived alleles increased the intensity of black, and red or yellow pigmentations, respectively. The only QTL for pericarp color co located with the red pericarp gene Rc, with the SS18-2-derived allele increasing the intensity of black and red pigmentations. Four of the five dormancy QTLs were flanked or bracketed by one to four QTLs for the interrelated characteristics. The QTL organization pattern indicates the central role of seed dormancy in adaptive syndromes for non-domesticated plants, implies that the elimination of dormancy from cultivars could arise from the selections against multiple interrelated characteristics, and challenges the use of dormancy genes at these loci in breeding varieties for resistance to pre-harvest sprouting (PHS). However, another QTL (qSD12) provides candidate gene(s) for PHS resistance because it has a large effect in the population and it is independent of the loci for interrelated characteristics. PMID- 15782298 TI - Cystatin C: our experience. PMID- 15782299 TI - Renal function during and after treatment for acute lymphoblastic leukemia in children. AB - Renal function tests (cystatin C, serum and urine creatinine, creatinine clearance, serum and urine beta(2)-microglobulin, microalbuminuria, osmolality) were performed in 21 children at the diagnosis and during the treatment for acute lymphoblastic leukemia (ALL) (group I) and in 37 children (group II) treated for ALL 3.9+/-3.7 years before the study. The results were compared to 20 healthy children. Mean values of renal tests were in normal range at all points of analysis in groups I and II compared to the control group. Transitory higher cystatin C values (but in normal range) were observed after methotrexate administration and after the end of treatment. Deteriorated renal function was observed in one child during the treatment (after each protocol) and in five children treated previously for ALL. In conclusion, combined treatment for ALL is not associated with severe or long-term impairment of renal function. PMID- 15782300 TI - Hypocitraturia as a risk factor for nephrocalcinosis after kidney transplantation. AB - Calcium-oxalate crystal deposition in kidney transplant biopsy specimen led us to investigate the impact of calcineurin inhibitor treatment on urinary excretion of lithogenic and stone inhibitory substances in 53 children after successful kidney transplantation (KTx) receiving cyclosporine A (CsA) or tacrolimus. We compared the values obtained with those of 12 patients with recurrent nephrotic syndrome under CsA and of 6 patients with Rasmussen encephalitis (RE) under tacrolimus therapy. Renal ultrasound examinations were repeatedly performed. Hypocitraturia was found in 69% of patients, with KTx patients having a significantly lower urinary citrate excretion than those receiving calcineurin inhibitors for other reasons. Secondly, we found hyperoxaluria in 35% of patients, again especially in those after KTx. No significant difference in urinary substances was seen comparing CsA with tacrolimus treatment. Urolithiasis was found in one and calcium-oxalate crystal deposition in biopsy specimen of three KTx patients. Calcineurin inhibitor treatment can lead to significant hypocitraturia, especially in patients after KTx receiving the highest dose of medication. Hyperoxaluria is primarily the result of a removal of significant body oxalate stores, deposited during dialysis, but may not be suspected as a specific side effect of calcineurin inhibitor therapy. Both findings can increase the risk for urolithiasis or nephrocalcinosis. PMID- 15782301 TI - Long-term renal follow-up of extremely low birth weight infants. AB - There is evidence that low birth weight caused by intrauterine growth retardation adversely affects normal renal development. Very little information on this issue is available on children born very prematurely. This investigation examined clinical and functional renal parameters in 40 children (23 boys, 17 girls) ranging in age between 6.1 and 12.4 years and weighing less than 1000 g at birth. Results were compared to those obtained in 43 healthy children of similar age and gender. Study subjects were significantly smaller and thinner than control subjects (mean height SDS: -0.36 vs. +0.70; and mean BMI SDS: -0.56 vs. +1.18). Systolic, diastolic, and mean blood pressures did not differ from those of controls. Renal sonography revealed no abnormality, and mean percentiles for renal length and volume appeared normal. In comparison with controls, plasma creatinine concentration (0.62+/-0.1 vs. 0.53+/-0.1 mg/dl) and estimated creatinine clearance (117+/-17 vs. 131+/-17 ml min(-1) 1.73 m(-2)) differed significantly. No significant differences were observed in microalbuminuria values, but five study subjects (12.5%) presented values above the upper limit of normality. A defect in tubular phosphate transport was also evident: TmP/GFR (3.6+/-0.4 vs. 4.2+/-0.8 mg/dl) and TRP (83+/-5% vs. 90+/-4%) were significantly lower, and urinary P excretion, estimated by the ratio UP/UCr, was significantly higher (1.2+/-0.4 vs. 0.9+/-0.4 mg/mg) than controls. Urinary calcium excretion, estimated by the UCa/UCr ratio, was also significantly higher (0.15+/-0.07 vs. 0.12+/-0.09 mg/mg). These data clearly demonstrate that both GFR and tubular phosphate transport are significantly diminished in school-age children born with extreme prematurity, probably as a consequence of impaired postnatal nephrogenesis. PMID- 15782302 TI - Long-term follow-up of diffuse membranoproliferative glomerulonephritis type I. AB - In Japan, the school urinary screening system facilitates early detection and treatment of membranoproliferative glomerulonephritis (MPGN) in childhood. The present study investigated the long-term prognosis in 19 children with diffuse MPGN type I who received steroid therapy. Before signs of glomerulonephritis were confirmed, all patients displayed abnormal urinalysis results, predominantly through school urinary screening. Treatment comprised a regimen of alternate-day prednisolone after steroid pulse or cyclophosphamide therapy, and follow-up was continued for 10-24 years. Excluding 1 patient on short-term therapy, 18 patients received long-term alternate-day prednisolone therapy for 4-12 years. Treatment was discontinued when amelioration was confirmed on renal biopsy. As of the last observation, urinary abnormalities and hypocomplementemia had disappeared in 15 patients, while mild proteinuria without hypocomplementemia remained in 4 patients. No patients required hemodialysis. Moreover, no severe adverse effects attributable to treatment were identified other than mild short stature. Early detection and therapy using pulse methylprednisolone followed by alternate-day prednisolone was thus confirmed as safe and useful for treating diffuse MPGN type I. PMID- 15782303 TI - 9alpha-Fluorohydrocortisone therapy in aldosterone synthase deficiency. PMID- 15782304 TI - Factitious proteinuria fabricated with adding human albumin: how to detect it? PMID- 15782305 TI - Hypocomplementaemia and septicaemia. PMID- 15782306 TI - Heat-shock protein 70: molecular supertool? AB - The cellular stress response decreases cellular injury, either via primary induction of cytoresistance or by secondary enhancement of cellular repair mechanisms. The most frequently studied and best understood effectors of the cellular stress response are the heat shock proteins (HSP). HSP are among the oldest tools in the cellular protein machinery, demonstrating extremely high conservation of the genetic code since bacteria. Molecular chaperons, with the HSP-70 being the prototype, cooperate in transport and folding of proteins, preventing aggregation, and even resolubilizing injured proteins. Increasing evidence supports a role for HSP during the recovery from renal ischemia, in particular in cellular salvage from apoptotic cell death and cytoskeletal restoration. Recent studies also report the potential for biomolecular profiling of newborns for the risk of acute renal failure. In peritoneal dialysis novel data suggest the use of HSP expression for biocompatibility testing. More importantly, HSP are prime therapeutic candidates for clinical situations associated with predictable insults, such as organ procurement in transplant medicine and repetitive exposure to hyperosmolar and acidotic peritoneal dialysis fluids. The next challenge will be to define the regulatory pathways of the cellular stress response in these models to introduce novel therapeutic interventions, such as new pharmaceutics enhancing the HSP expression. PMID- 15782307 TI - Dysregulation of renal MMP-3 and MMP-7 in canine X-linked Alport syndrome. AB - Matrix metalloproteinases (MMPs) play an important regulatory role in many biological and pathological processes and their specific role in Alport syndrome (AS) is not yet clearly defined. In this study, the naturally occurring canine X linked AS was used to demonstrate a potential role for MMP-3 and MMP-7 in Alport renal pathogenesis. Recently, we demonstrated that the expression of MMP-2, MMP-9 and MMP-14 was upregulated in the renal cortex of dogs with a spontaneous form of XLAS. In the present study, we examined necropsy samples of renal cortex from normal and XLAS dogs for MMP-3 and MMP-7 as they have the potential to activate MMP-2 and MMP-9. Immunohistochemical analysis showed strong immunostaining for both MMP-3 and MMP-7 in the interstitial space of XLAS kidneys, while virtually no immunostaining was observed in similar fields from normal dogs. RT-PCR and casein zymography confirmed that both mRNA transcripts and activities of MMP-3 and MMP-7 are elevated in XLAS kidneys. The induction of these MMPs likely contributes to tissue destruction associated with the fibrogenic process, while augmenting the activation of MMP-2 and MMP-9 by MMP-3 and MMP-7 in XLAS. Thus, these data further implicate a role for the MMPs in progressive renal pathogenesis associated with AS. PMID- 15782308 TI - Growth hormone for children with chronic renal failure and on dialysis. AB - We studied all children with CRF who received recombinant human growth hormone (rhGH) for more than a year (mean+/-SD duration of therapy 3.7+/-2.5 years) over an 11-year period. There were 32 children. Twenty-one children were conservatively managed, with a mean glomerular filtration rate (GFR) of 24+/-12 mL min(-1)/1.73 m2 at the start of rhGH. Their height standard deviation score improved from -2.5+/-1.4 to -2.1+/-0.7 at 1 year (P=0.3), -2.0+/-0.7 at 2 years (P=0.01), and -1.6+/-0.6 at 3 years (P=0.001). After that there was no improvement. Eleven children were on dialysis, six on haemodialysis (HD) and five on peritoneal (PD). Ht SDS improved from -2.7+/-0.5 to -2.3+/-0.5 at 1 year (P=0.02). Thereafter there was no further improvement. RhGH was stopped because of transplantation in 29 patients at a mean+/-SD age of 12.1+/-4.0 years. Mean Ht SDS was -1.8+/-0.8 at transplant and there was no change over the following 5 years. In conclusion, treatment with rhGH resulted in improvement in Ht SDS in conservatively managed CRF for up to 3.0 years and for 1 year in children on dialysis. Discontinuation of rhGH after transplantation resulted in little change in Ht SDS. PMID- 15782309 TI - Blood pressure in the long-term follow-up of children with haemolytic uremic syndrome. PMID- 15782311 TI - Anti-GBM and anti-MPO antibodies coexist in a case of pulmonary renal syndrome. AB - The case of a 12-year-old boy with pulmonary renal syndrome is described. Antimyeloperoxidase (anti-MPO) and antiglomerular basement membrane (anti-GBM) antibodies were positive. The clinical course and immunosuppressive therapy are discussed. Pulmonary renal syndrome is a rare event in childhood and coexistence of the two types of antibodies is exceptional. PMID- 15782310 TI - Serum and urine leptin concentration in children with nephrotic syndrome. AB - Literature data point to the relationship between leptin concentration and certain markers of the metabolic syndrome, including cholesterol, triglycerides and apolipoproteins. A substantial lipid metabolism disturbance occurs in children with idiopathic nephrotic syndrome (NS). The aim of the study was to find out whether in NS children, serum and urine leptin levels change proportionally to lipid metabolism disturbances. The study was performed on two groups: (I) 30 children with NS (A) before, (B) during, prednisone therapy after proteinuria regression; (II) 25 healthy children. Serum and urine leptin levels were determined by the immunoenzymatic ELISA method. Serum leptin level in NS children before and after treatment was similar to that in the control group (p>0.05). Leptin urinary excretion in group A was approximately 60 times and in group B 24 times higher than in the controls (p<0.01). Before treatment, children with NS had increased concentrations of TC, TG, LDL, beta-lipoprotein, apolipoprotein B (apo B) (p<0.01) and reduced HDL and apolipoprotein A (apo A) (p<0.01). The conclusions were that: (1) in NS children leptin urinary excretion increases but its level is unchanged in serum; (2) serum leptin level is correlated with lipid parameters. PMID- 15782312 TI - Effect of gemcitabine on immune cells in subjects with adenocarcinoma of the pancreas. AB - Effects of gemcitabine (Gemzar) on immune cells were examined in pancreas cancer patients to determine whether it was immunosuppressive, or potentially could be combined with vaccines or other immunotherapy to enhance patient's responses to their tumors. Blood was obtained at five time-points, before therapy, 3-4 days after initial gemcitabine infusion and immediately preceding three additional weekly infusions. Effects on T-cell subsets, B-cells, myeloid dendritic cell precursors, antigen presenting cells (APC), activated/memory, and naive cells were examined. Functional activity was measured by intracellular staining for cytokines before and after T-cell activation, and by interferon gamma production in EliSpot responses to tumor presentation. Although absolute lymphocyte counts decreased with the initial treatment with gemcitabine infusion, the counts stabilized during subsequent treatments, then returned within normal ranges seven days after the fourth treatment so that the absolute lymphocyte count no longer differed significantly from that prior to treatment. These effects on absolute lymphocyte counts were mirrored by statistically significant decreases in absolute numbers of CD3 and CD20 lymphocytes during these time periods. The proportions of T and B-cells, however did not change significantly with therapy, although significance changes were observed in some specialized subsets. A decrease in the proportions of the major BDCA-1+, CD1b myeloid dendritic cell subset and a reciprocal increase in the minor BDCA-3+ dendritic cell subsets resulted at 3-4 days, then their levels returned to normal. No significant changes in percentages of CD86 and CD80 APCs or CD4+, CD25+ T-cells were documented. Increased percentages of CD3+, CD45RO+ memory lymphocytes reached significance at day 7, then declined to statistically significant decrease at days 14 and 21 after the second and third infusions, respectively. Immune T-cells were functional in pancreas cancer patients treated with gemcitabine. The data suggest that gemcitabine therapy may decrease memory T-cells and promote naive T cell activation. We conclude that gemcitabine therapy (1) is not immunosuppressive and (2) may enhance responses to specific vaccines or immunotherapy administered to activate or support immune responses directed toward driving effector immunity to cancer cells. PMID- 15782313 TI - Mitomycin-C and capecitabine as third-line chemotherapy in patients with advanced colorectal cancer: a phase II study. AB - PURPOSE: The aim of this study was to investigate the therapeutic value and safety of third-line treatment with mitomycin-C (MMC) and capecitabine (Xeloda) in patients with advanced colorectal cancer pretreated with combination regimens including 5-fluorouracil (5-FU), folinic acid (FA) and irinotecan (CPT-11) or 5 FU, FA and oxaliplatin (L-OHP). PATIENTS AND METHODS: A total of 21 patients (M/F 16/5, median age 60.0 years) with advanced colorectal cancer, all of whom had developed progressive disease while receiving or within 6 months of discontinuing two sequential chemotherapy lines with 5-FU, FA and CPT-11 or 5-FU, FA and L-OHP, were accrued to this study. At the time of their relapse or progression, cytotoxic chemotherapy, consisting of intravenous MMC 7 mg/m(2) on therapeutic day 1 plus oral capecitabine 1000 mg/m(2) twice daily on days 1-14, was initiated. After rest for 7 days, capecitabine 1000 mg/m(2) twice daily was administered on days 22-35 followed by 7 days rest. Treatment courses were repeated every 6 weeks unless there was evidence of progressive disease, unacceptable toxicity or patient refusal of treatment. RESULTS: All the patients were assessable for toxicity and 19 for response. The median number cycles of chemotherapy was two (range one to four). Only 1 patient (4.8%) had a partial response, 4 patients (19.0%) had stable disease, and 14 patients (66.7%) progressed. The median follow-up period was 7.3 months and median time to progression was 2.6 months. The median overall survival was 6.8 months. No toxic deaths occurred. Toxicities of third-line treatment were mild and manageable. As NCI/NIH common toxicity criteria, grade 3/4 anemia, neutropenia and thrombocytopenia occurred in two, one and one patients, respectively. CONCLUSION: Our findings suggest that the combination of MMC and capecitabine in patients with advanced colorectal cancer pretreated with combination regimens including 5 FU, FA and CPT-11 or 5-FU, FA and L-OHP is safe. However, this regimen had a poor response rate and no definitive contribution to increasing patients' overall survival time. Further evaluation of other salvage regimens seems to be warranted. PMID- 15782314 TI - Cerebral aspergillosis in the critically ill: two cases of successful medical treatment. AB - OBJECTIVE: Invasive aspergillosis is associated with a poor prognosis, especially in critically ill patients with cerebral involvement. We present two cases of cerebral invasive aspergillosis successfully treated in the intensive care unit with combination antifungal therapies and without surgery. CASE PRESENTATION: The first patient was a 49-year-old man with rheumatoid arthritis who received corticosteroid and cyclophosphamide treatment and developed pulmonary and cerebral invasive aspergillosis. After failure of voriconazole the patient had a successful outcome with voriconazole and liposomal amphotericin B therapy. The patient returned home after an 8-month hospital stay. The second patient was a 54 year-old woman with pulmonary neoplasia and corticosteroid treatment who developed pulmonary and cerebral invasive aspergillosis. After failure of voriconazole and liposomal amphotericin B therapy the patient had a favorable outcome with liposomal amphotericin B and caspofungin therapy. The patient died 10 months after initial diagnosis of cardiac tamponade unrelated to fungal infection. DISCUSSIONS: These cases illustrate the improving prognosis of invasive aspergillosis due to the availability of new treatments, especially in cases of cerebral involvement. It also demonstrates that the outcome of critically ill patients requiring mechanical ventilation for invasive aspergillosis can be favorable. The treatment of patients with invasive cerebral aspergillosis in the intensive care setting should be encouraged. PMID- 15782316 TI - Documented and clinically suspected bacterial infection precipitating intensive care unit admission in patients with hematological malignancies: impact on outcome. AB - OBJECTIVE: To assess the impact of documented and clinically suspected bacterial infection precipitating ICU admission on in-hospital mortality in patients with hematological malignancies. DESIGN AND SETTING: Prospective observational study in a 14-bed medical ICU at a tertiary university hospital. PATIENTS: A total of 172 consecutive patients with hematological malignancies admitted to the ICU for a life-threatening complication over a 4-year period were categorized into three main groups according to their admission diagnosis (documented bacterial infection, clinically suspected bacterial infection, nonbacterial complications) by an independent panel of three physicians blinded to the patient's outcome and C-reactive protein levels. RESULTS: In-hospital and 6-months mortality rates in documented bacterial infection (n=42), clinically suspected bacterial infection (n=40) vs. nonbacterial complications (n=90) were 50.0% and 42.5% vs. 65.6% (p=0.09 and 0.02) and 56.1% and 48.7% vs. 72.1% (p=0.11 and 0.02), respectively. Median baseline C-reactive protein levels in the first two groups were 23 mg/dl and 21.5 mg/dl vs. 10.7 mg/dl (p<0.001 and p=0.001) respectively. After adjustment for the severity of critical and underlying hematological illness and the duration of hospitalization before admission documented (OR 0.20; 95% CI 0.06 0.62, p=0.006) and clinically suspected bacterial infection (OR 0.18; 95% CI 0.06 0.53, p=0.002) were associated with a more favorable outcome than nonbacterial complications. CONCLUSIONS: Severely ill patients with hematological malignancies admitted to the ICU because of documented or clinically suspected bacterial infection have a better outcome than those admitted with nonbacterial complications. These patients should receive advanced life-supporting therapy for an appropriate period of time. PMID- 15782315 TI - Bile modulates intestinal epithelial barrier function via an extracellular signal related kinase 1/2 dependent mechanism. AB - OBJECTIVE: Obstructive jaundice is frequently complicated by infections and has been associated with increased bacterial translocation and gut mucosal hyperpermeability in animal models. Proper expression of the tight junction (TJ) proteins ZO-1 and occludin is important for normal gut barrier function. We tested whether bile modulates intestinal epithelial ZO-1 and occludin expression. ANIMALS: (a) Male C57BL/6 mice; (b) male Sprague-Dawley rats. INTERVENTIONS: (a) Mice were subjected to common bile duct ligation (CBDL) or a sham procedure, and 96 h later all surviving animals were killed for measurement of ileal mucosal permeability to FITC-labeled dextran (everted gut sac technique), bacterial translocation to mesenteric lymph nodes, and ileal epithelial ZO-1 and occludin expression (western blots). (b) Rat IEC-6 enterocytic monolayers were incubated in the presence or absence of graded concentrations of rat bile and/or U0126, an inhibitor of extracellular signal related kinase (ERK) 1/2 activation. RESULTS: (a) Compared to sham-treated controls, CBDL significantly increased gut mucosal permeability and bacterial translocation and markedly decreased ileal epithelial expression of ZO-1 and occludin. In a follow-up in vivo experiment, gavaging mice with fresh rat bile twice daily significantly ameliorated the deleterious effects of CBDL on gut barrier function. (b) Addition of 1% (v/v) bile to media enhanced phosphorylation of ERK1/2, increased the expression of ZO-1 and occludin and decreased permeability to FITC-dextran. All of these bile-mediated effects were blocked by 10 microM U0126. CONCLUSIONS: These data support the view that the presence of bile in the intestinal lumen is essential for normal gut barrier function, possibly because compounds present in bile initiate ERK1/2-dependent signaling that is essential for normal expression of key TJ proteins. PMID- 15782317 TI - Modeling MODS: what can be learned from animal models of the multiple-organ dysfunction syndrome? PMID- 15782319 TI - Failure of new antifungals to control Candida thrombophlebitis. PMID- 15782318 TI - A pilot study of inhaled nitric oxide in preterm infants treated with nasal continuous positive airway pressure for respiratory distress syndrome. AB - OBJECTIVE: To explore the acute effects of inhaled nitric oxide (iNO) on oxygenation, respiratory rate, and CO2 levels in spontaneously breathing preterm infants treated with nasal continuous positive airway pressure (nCPAP) for moderate respiratory distress syndrome (RDS). DESIGN AND SETTING: Randomized, prospective, double-blind, cross-over study in the neonatal intensive care units of a university hospital. PATIENTS: 15 infants treated for RDS, with a median gestational age of 32 weeks (27-36), birth weight 1940 g (1100-4125), and postnatal age at the beginning of study 23 h (3-91). nCPAP pressure was kept constant at 4.3 cmH2O (3.4-5.1). INTERVENTIONS: We examined effects on gas exchange and vital signs during a 30-min exposure to 10 ppm iNO or placebo gas (nitrogen). RESULTS: Before administering test gases the baseline arterial to alveolar oxygen tension ratio (aAPO2) was 0.19+/-0.06. aAPO2 remained unchanged during placebo but increased to 0.22+/-0.05 (+20%) during iNO exposure. Respiratory rate and arterial carbon dioxide tension remained unchanged, as did heart rate, blood pressure, and methemoglobin. Follow-up at 30 days of age showed no deaths, delayed morbidity, or need for supplemental oxygen. CONCLUSIONS: Adding 10 ppm nitric oxide to nasal CPAP treatment in preterm infants suffering from RDS results in a moderate but statistically significant improvement in oxygenation, with no effect on respiratory drive or systemic circulatory parameters. PMID- 15782320 TI - [Scientific findings and clinical practice. The story of the hare and hedgehog]. PMID- 15782321 TI - FMRFamide-like immunoreactivity in the sea-fan Eunicella cavolini (Cnidaria: Octocorallia). AB - The presence of FMRFamide-related peptides (FaRPs) was investigated, by immunohistochemical methods with a polyclonal FMRFamide antiserum, in the sea-fan Eunicella cavolini (Van Koch 1887), a representative of the cnidarians (octocorallians). The identification of FaRP-immunoreactive elements as neuronal cells and a nerve net was performed by double immunohistochemical methods with the monoclonal anti-beta-tubulin antibody. A strong and widely distributed FaRPs immunoreactivity was detected: FaRPs-immunoreactive nerve cells were observed among and underlying gastrodermal epithelial cells, epidermal cells lining tentacles, muscular septs and gonophores. A diffuse FaRPs-immunoreactive nerve net was also found between epithelia and mesoglea and in the stalk of the gonophore. These results improve our knowledge of the gorgonian nervous system and demonstrate that most of the immunoreactive cells belong to neural elements. PMID- 15782322 TI - Cell-death mechanisms in the IPLB-LdFB insect cell line: a nuclear located Bcl-2 like molecule as a possible controller of 2-deoxy-D-ribose-mediated DNA fragmentation. AB - In the IPLB-LdFB insect cell line, oncosis and apoptosis are the two pre-mortal processes, whereas necrosis is the post-mortem condition. As found in mammals, adenosine triphosphate depletion of insect cells by oligomycin A induces oncosis. The apoptotic inducer 2-deoxy-D: -ribose (dRib) provokes cell death through an intrinsic apoptotic pathway similar to that observed in mammalian models and results in oligonucleosomal DNA fragmentation. The addition to insect cells of an anti-Bcl-2 polyclonal antibody known to prevent dRib-mediated apoptosis abolishes DNA fragmentation, whereas cytochrome c release and the increase in a caspase 3 like activity are still detectable. These and previous findings suggest a double role for the Bcl-2-like molecule in IPLB-LdFB, i.e. the maintenance of mitochondrial integrity and the control of apoptotic machinery at the nuclear level. PMID- 15782323 TI - mRNA for pancreatic uncoupling protein 2 increases in two models of acute experimental pancreatitis in rats and mice. AB - Uncoupling-protein 2 (UCP2) is a mitochondrial protein that appears to be involved in cellular oxidant defense and in the regulation of oncotic cell death, both of which are important features of acute pancreatitis. However, UCP2 expression in acute pancreatitis has not been previously reported. In the current experiments, pancreatic gene expression was studied by real-time reverse transcription/polymerase chain reaction and Northern blots. Two models of acute experimental pancreatitis were investigated: cerulein-induced pancreatitis in mice at two different time points and taurocholate-induced pancreatitis in rats at two degrees of severity. After cerulein administration, acinar injury and leukocyte infiltration was significantly higher at 24 h compared with 12 h after the first injection of cerulein (P<0.05, P<0.005, respectively). UCP2 mRNA was unchanged at 12 h but was nearly 12-fold greater than control levels after 24 h (P<0.001). UCP2 gene expression correlated with acinar injury (r=0.69; P<0.001). By 72 h after taurocholate administration, the severe group had more necrosis than the mild group (P<0.005). Pancreatic UCP2 mRNA was increased fourfold in the severe group compared with controls (P<0.01). UCP2 expression correlated with parenchymal necrosis (r=0.61; P<0.01). Thus, pancreatic UCP2 mRNA increased in two models of acute pancreatitis. The increase in UCP2 gene expression was correlated with the severity of the disease. Up-regulation of UCP2 in the pancreas may be a protective response to oxidative stress, but this increase may also have a negative influence on cellular energy metabolism. Therefore, acinar UCP2 may be an important modifier of the severity of acute pancreatitis. PMID- 15782324 TI - [Augmenting atypical antipsychotic medications with clozapin]. AB - Typical antipsychotic medications have considerably improved clinical outcome of patients suffering from schizophrenic psychoses, but up to 40% of the cases show treatment resistant symptoms. Even therapy with atypical antipsychotic drugs such as risperidone, quetiapine, olanzapine, sulpiride, amisulpride, and ziprasidone often fails to reach complete remission due to resistant, positive or negative symptoms or dose-limiting side effects. As this also holds true in the case of monotherapy with clozapine, a substance known to be effective against treatment resistant schizophrenia, increasing numbers of patients receive atypical antipsychotic drugs in addition to clozapine. This review systematically evaluates case reports and clinical investigations on the use of clozapine combined with risperidone, olanzapine, quetiapine, sulpiride, amisulpride, or ziprasidone. Details on indication, methodology, and effects of the investigations are summarized. Only one double blind, placebo-controlled trial on the combination with sulpiride exists within a number of altogether 31 publications about 1182 treatments. Favorable effects on positive and/or negative symptoms or improvements of clozapine-induced side effects were described for every combination approach. In some cases pharmacokinetic interactions or serious unfavorable effects occurred. In conclusion it might be accepted that most of the combination therapies follow a neurobiological rational. There a major differences in the level of evidence that they are safe, tolerable and effective. We discuss criteria for the indication for augmenting clozapine therapy and the differential indication for existing alternatives. Additional randomized prospective trials are needed in order to evaluate these strategies systematically. PMID- 15782326 TI - The GC-MS detection and characterization of neopine resulting from opium use and codeine metabolism and its potential as an opiate-product-use marker. AB - Neopine, a minor opium alkaloid and an isomer of codeine (also known as beta codeine), has been detected in both the urine of opium users and pharmaceutical codeine users. The characterization of neopine was achieved by comparison of the mass spectra and GC retention times of the trimethylsilyl derivative. The presence of neopine in the urine of pharmaceutical codeine users was attributed to the metabolism of codeine through a double bond migration in ring C, from the 7-8 to the 8-14 position. The potential use of the alkaloid as a confirmation marker of opium and/or pharmaceutical codeine use and the ability to differentiate these from heroin use has been discussed. PMID- 15782327 TI - Metal components analysis of metallothionein-III in the brain sections of metallothionein-I and metallothionein-II null mice exposed to mercury vapor with HPLC/ICP-MS. AB - Mercury vapor is effectively absorbed via inhalation and easily passes through the blood-brain barrier; therefore, mercury poisoning with primarily central nervous system symptoms occurs. Metallothionein (MT) is a cysteine-rich metal binding protein and plays a protective role in heavy-metal poisoning and it is associated with the metabolism of trace elements. Two MT isoforms, MT-I and MT II, are expressed coordinately in all mammalian tissues, whereas MT-III is a brain-specific member of the MT family. MT-III binds zinc and copper physiologically and is seemed to have important neurophysiological and neuromodulatory functions. The MT functions and metal components of MTs in the brain after mercury vapor exposure are of much interest; however, until now they have not been fully examined. In this study, the influences of the lack of MT-I and MT-II on mercury accumulation in the brain and the changes of zinc and copper concentrations and metal components of MTs were examined after mercury vapor exposure by using MT-I, II null mice and 129/Sv (wild-type) mice as experimental animals. MT-I, II null mice and wild-type mice were exposed to mercury vapor or an air stream for 2 h and were killed 24 h later. The brain was dissected into the cerebral cortex, the cerebellum, and the hippocampus. The concentrations of mercury in each brain section were determined by cold vapor atomic absorption spectrometry. The concentrations of mercury, copper, and zinc in each brain section were determined by inductively coupled plasma mass spectrometry (ICP-MS). The mercury accumulated in brains after mercury vapor exposure for MT-I, II null mice and wild-type mice. The mercury levels of MT-I, II null mice in each brain section were significantly higher than those of wild-type mice after mercury vapor exposure. A significant change of zinc concentrations with the following mercury vapor exposure for MT-I, II null mice was observed only in the cerebellum analyzed by two-way analysis of variance. As for zinc, the copper concentrations only changed significantly in the cerebellum. Metal components of metal-binding proteins of soluble fractions in the brain sections were analyzed by size exclusion high-performance liquid chromatography (HPLC) connected with ICP-MS. From the results of HPLC/ICP-MS analyses, it was concluded that the mercury components of MT-III and high molecular weight metal-binding proteins in the cerebellum of MT-I, II null mice were much higher than those of wild-type mice. It was suggested that MT-III is associated with the storage of mercury in conditions lacking MT-I, and MT-II. It was also suggested that the physiological role of MT-III and some kind of high molecular weight proteins might be impaired by exposure to mercury vapor and lack of MT-I and MT-II. PMID- 15782329 TI - Rapid and sensitive method for the determination of acetaldehyde in fuel ethanol by high-performance liquid chromatography with UV-Vis detection. AB - A high-performance liquid chromatography (HPLC) method for the determination of acetaldehyde in fuel ethanol was developed. Acetaldehyde was derivatized with 0.900 mL 2,4-dinitrophenylhydrazine (DNPHi) reagent and 50 microL phosphoric acid 1 mol L(-1) at a controlled room temperature of 15 degrees C for 20 min. The separation of acetaldehyde-DNPH (ADNPH) was carried out on a Shimadzu Shim-pack C18 column, using methanol/LiCl((aq)) 1.0 mM (80/20, v/v) as a mobile phase under isocratic elution and UV-Vis detection at 365 nm. The standard curve of ADNPH was linear in the range 3-300 mg L(-1) per injection (20 microL) and the limit of detection (LOD) for acetaldehyde was 2.03 microg L(-1), with a correlation coefficient greater than 0.999 and a precision (relative standard deviation, RSD) of 5.6% (n = 5). Recovery studies were performed by fortifying fuel samples with acetaldehyde at various concentrations and the results were in the range 98.7 102%, with a coefficient of variation (CV) from 0.2% to 7.2%. Several fuel samples collected from various gas stations were analyzed and the method was successfully applied to the analysis of acetaldehyde in fuel ethanol samples. PMID- 15782330 TI - Leadership. PMID- 15782332 TI - Miniaturisation and automation of metal fractionation schemes applied to environmental solid samples by sequential injection microcolumn extraction procedures. PMID- 15782331 TI - Determination of HCN sampled from gasification product gases by headspace gas chromatography with atomic emission detector. AB - Nitrogen-containing fuels produce hydrogen cyanide when the fuel is gasified. The gas is poisonous and produces nitrogen oxides when it is burned. HCN is usually sampled into alkaline solutions and analysed using an ion selective electrode. The method is tedious and the electrode response is temperature-dependent. Samples are not stable and must be analysed immediately, and they contain ions which are poisonous to the electrode. Therefore a new gas chromatographic method was developed. In this new method HCN is released from the alkaline solutions with sulphuric acid in a headspace sampler and analysed by a gas chromatograph connected to an atomic emission detector. Measurements on carbon emission line 193.1 nm gave the limit of detection 0.05 mg CN-/l in the solution. The calibration curve was linear to 1000 mg CN-/l and the correlation was 0.997. The relative standard deviation of the calibration was 1.7% at the concentration of 5 mg CN-/l and 1.0% at 25 mg CN-/l. The developed headspace method allows automated analysis and it needs less sample preparations than the ion selective electrode method. This paper also reports the effect of sample preparation and storage time on the stability of the samples. PMID- 15782333 TI - Copper, nickel, and vanadium in the Western Galician Shelf in early spring after the Prestige catastrophe: is there seawater contamination? AB - The original cargo fuel transported by the Prestige tanker, according to our analysis, contained several trace elements, for example Cu (3 microg g(-1)), Ni (97 microg g(-1)), and V (382 microg g(-1)). The possibility of metal contamination of seawater near the bottom from the sedimented fuel and in surface waters from the fuel deposited on the littoral area was researched in the Western Galician Shelf 5 months after the Prestige shipwreck (November 2002). Water samples obtained at eight stations from Finisterre Cape to Mino River were analysed by means of an electroanalytical method (AdCSV) and following trace metal-clean procedures. Only the surface Cu and Ni levels at the Ons and Cies Islands were higher than those considered as background for Open North East Atlantic Ocean waters. The accumulation and weathering of the fuel deposited on the shores could be a significant source for the enrichment found, but the influence of industrial and urban inputs from the neighbouring Pontevedra and Vigo rias is also possible. Thus, this study was useful for establishing typical values of Cu, Ni, and V in Galician Shelf waters for the first time. Total metal concentrations ranged between 1.7 and 3.4 nmol L(-1) for Cu, between 2.9 and 4.8 nmol L(-1) for Ni, and between 19.7 and 32.3 nmol L(-1) for V; dissolved concentrations were between 0.7 and 1.5 nmol L(-1) for Cu, between 2.2 and 3.3 nmol L(-1) for Ni, and between 15.5 and 27.0 nmol L(-1) for V; particulate values below 2.6 nmol L(-1) for Cu, 2 nmol L(-1) for Ni, and 7.2 nmol L(-1) for V were obtained. PMID- 15782334 TI - Quantitative determination of cationic modified polysaccharides on hair using LC MS and LC-MS-MS. AB - Cationic polysaccharides containing N-hydroxypropyl-N,N,N-trimethylammonium substituents are widely used as conditioning agents for hair-care products. A sensitive method has been developed for the quantitation of these polymers. After acidic extraction from hair the polysaccharides are hydrolyzed using trifluoroacetic acid. The cationic monoglycosides are determined using liquid chromatography-tandem mass spectrometry (LC-MS-MS). The developed method is independent of hair treatment. Even hair cut from test persons after customary hair wash can be analyzed. After treatment of natural and bleached hair tresses using a real-life treatment procedure 180 microg and 300 microg of polymer per gram hair were quantified, respectively. Additionally the fragmentation mechanism of the cationic N-hydroxypropyl-N,N,N-trimethylammonium group during electrospray ionization was investigated. A mass loss of 60 Da in combination with loss of a single charge is observed and associated with cleavage of trimethylamine and a proton. It is assumed that this process is promoted by the anionic counter-ion which might be hydroxide in an aqueous environment. PMID- 15782335 TI - A kinetic model and its parameter estimation for the process of binding copper to human serum albumin by a voltammetric method. AB - Linear sweep anodic stripping voltammetry was applied to determine the concentration of free copper ions in the process of binding copper to human serum albumin (HSA) on the mercaptoethane sulfonate modified gold electrode surface. A kinetic model of two consecutive steps for the process of binding copper to HSA was first proposed on the basis of the electrochemical results and compared with a parallel kinetic response model by using residual analysis. The experimental data of the stripping peak currents with time was fitted according to the model and the kinetic parameters, binding rate constants, k1 and k2, were estimated to be 0.411 and 0.055 min(-1), respectively. PMID- 15782337 TI - Robust and non-parametric statistics in the evaluation of figures of merit of analytical methods. Practices for students. AB - A set of laboratory practices is proposed in which evaluation of the quality of the analytical measurements is incorporated explicitly by applying systematically suitable methodology for extracting the useful information contained in chemical data. Non-parametric and robust techniques useful for detecting outliers have been used to evaluate different figures of merit in the validation and optimization of analytical methods. In particular, they are used for determination of the capability of detection according to ISO 11843 and IUPAC and for determination of linear range, for assessment of the response surface fitted using an experimental design to optimize an instrumental technique, and for analysis of a proficiency test carried out by different groups of students. The tools used are robust regression, least median of squares (LMS) regression, and some robust estimators as median absolute deviation (m.a.d.) or Huber estimator, which are very useful as an alternatives to the usual centralization and dispersion estimators. PMID- 15782336 TI - Comparative study of atomic fluorescence spectroscopy and inductively coupled plasma mass spectrometry for mercury and arsenic multispeciation. AB - Mercury and arsenic are two elements of undoubted importance owing to their toxic character. Although speciation of these elements has been developed separately, in this work for the first time the speciation of As and Hg using two atomic fluorescence detectors in a sequential ensemble is presented. A coupling based on the combination of high-performance liquid chromatography (where mercury and arsenic species are separated) and two atomic fluorescence detectors in series, with several online treatments, including photooxidation (UV) and hydride generation, has allowed the determination of mercury and arsenic compounds simultaneously. The detection limits for this device were 16, 3, 17, 12 and 8 ng mL(-1) for As(III), monomethylarsinic acid, As(V), Hg2+ and methylmercury, respectively. This coupling was compared with an analogous one based on inductively coupled plasma-mass spectrometry (ICP-MS) detection, with detection limits of 0.7, 0.5, 0.8, 0.9 and 1.1 ng mL(-1), respectively. Multispeciation based on ICP-MS exhibits better sensitivity than the coupling based on tandem atomic fluorescence, but this second device is a very robust system and exhibits obvious advantages related to the low cost of acquisition and maintenance, as well as easy handling, which makes it a suitable system for routine laboratories. PMID- 15782338 TI - Identification of natural dyes used in works of art by pyrolysis-gas chromatography/mass spectrometry combined with in situ trimethylsilylation. AB - Samples of four natural dyes from different organic families--natural madder (anthraquinonoid), curcuma (curcuminoid), saffron (carotenoid) and indigo (indigotic)--were analysed using a new procedure based on pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), which includes the on-line derivatisation of the natural dyes using hexamethyldisilazane (HMDS). In addition, a previous procedure involving the addition of a 10% H2SO4 aqueous solution to the dye and further separation with ethyl acetate has been tested. This procedure enhances the sensitivity of the method by extracting the colouring compounds from the rest of the compounds present in the natural dye. Two possible derivatising reagents--HMDS and tetramethylammonium hydroxide (TMAH)--were compared in order to assess their effectiveness in the proposed method. Characteristic peaks from trimethylsilyl derivatives of alizarin, quinizarin, xanthopurpurin and purpurin were obtained for madder; peaks from safranal, isophorone and trimethylsilyl derivative of crocetin for saffron; peaks from 4-(4 hydroxy-3-methoxy)phenyl-3-buten-2-one and 4-(4-hydroxy-3-methoxy)phenyl-2 butanone, which are primary pyrolysis products of curcuma, and peaks from indole, 2-methylindole and 2,3-dihydroindol-2-one, which are primary pyrolysis products of indigo, among others, were obtained. The reported procedure leads to the unambiguous identification of the four studied dyes from solid samples formed by individual dyes. PMID- 15782339 TI - Analysis of organic acids in wines by Fourier-transform infrared spectroscopy. AB - Fourier-transform infrared (FTIR) spectroscopy has been a major point of development in many wine laboratories in recent years. It enables almost instant analysis of several properties of wine, usually with very good precision and accurate results. Nevertheless, validation procedures should not be forgotten and should be fully performed. Recovery experiments were performed by spiking wine samples with different amounts of organic acids (tartaric, malic, lactic, acetic and citric--the most prominent in wines). After FTIR analysis of the total acidity and of each organic acid concentration, recoveries were calculated. For total acidity recovery results were, in general, good and very close to 100% (64 111%). On the other hand, for individual organic acid concentrations, the recovery results were lower than 100% (11-73%) for all spiking additions. These results could be explained by spectroscopic interferences between the organic acids. Because they have similar infrared spectra, it is not easy to distinguish between them and, therefore, to achieve accurate calibration. When total acidity, with a different infrared spectrum from the other abundant compounds in the wine, was taken as a single property the recovery results were acceptable. PMID- 15782340 TI - Lipoma arborescens affecting multiple joints. AB - Lipoma arborescens is a rare benign intra-articular lesion of unknown etiology that usually involves the suprapatellar pouch of the knee joint. Clinically, the most common finding is a slow-growing painless swelling, accompanied by intermittent effusion of the joint. We report a case of a multifocal lipoma arborescens localized in the knees and the hips in a 24-year-old man, initially mimicking an inflammatory arthropathy. The diagnosis of lipoma arborescens was made by magnetic resonance imaging of the hips and the knees. Under arthroscopic guidance, the synovial biopsy of the right knee disclosed the specific histological signs of lipoma arborescens. As far as we know, this is the third case of multifocal lipoma arborescens reported in the English literature. PMID- 15782341 TI - Osteochondritis dissecans after rotational acetabular osteotomy for dysplastic hip. AB - We encountered a rare case of osteochondritis dissecans of the femoral head after rotational acetabular osteotomy that recovered with conservative treatment 4 years after the detection of characteristic radiological findings. PMID- 15782342 TI - Stress-induced Salter-Harris I growth plate injury of the proximal tibia: first report. AB - We describe a case of chronic Salter-Harris I injury of the proximal tibial epiphysis. To our knowledge such an injury has not been described in the English language literature. The radiological appearance can mimic chronic infection. The possibility of chronic athletic stress-related change should be considered in such scenarios to avoid a misdiagnosis. PMID- 15782344 TI - Myelodysplastic syndrome of donor origin subsequent to successful treatment of myeloid/NK-cell precursor leukaemia with allogeneic PBSCT: two very rare conditions in one patient. AB - We report a 36-year-old male with myeloid/natural killer (NK)-cell precursor acute leukaemia with a complex aberrant karyotype, who was treated according to an acute-myeloid-leukaemia (AML) treatment protocol (idarubicine, cytarabine, and etoposide) followed by high-dose cytarabine consolidation and achieved complete remission. He underwent allogeneic matched unrelated donor (MUD) peripheral blood stem-cell transplantation (PBSCT) and remained in remission throughout his remaining life. Seven months posttransplantation, a myelodysplastic syndrome (MDS) with (20q-) of donor origin was diagnosed causing severe thrombocytopenia and finally leading to infection and death. This patient represents one of the few cases published achieving remission for a significant period of time after being diagnosed with myeloid/NK-cell precursor acute leukaemia, a very rare malignant disease. We conclude, despite the fatal outcome due to infection, that allogeneic PBSCT is a therapeutic option for patients with this entity. In addition, the development of a myelodysplastic syndrome of donor origin is extremely rare and only very few cases are published worldwide. PMID- 15782343 TI - Intensive chemotherapy with mitoxantrone administered as a single injection in patients with high-risk acute myeloid leukemia: results of the EMA 2000 trial. AB - Following a dose-escalation study performed to assess the maximally tolerated dose of high-dose mitoxantrone in a single injection combined with chemotherapy, a phase II trial (EMA 2000 regimen) was performed in patients with refractory or relapsed acute myelogenous leukemia (AML) between October 2000 and December 2003. Sixty-two patients entered the study and received mitoxantrone 45 mg/m(2) on day 1 in combination with cytarabine and etoposide. Overall, 39 patients (63%) achieved complete remission (CR). Four patients died during remission induction, and 19 patients had resistant disease. Median time to granulocyte and platelet recovery was 34 and 39 days, respectively. The predominant non-hematologic toxicity was infection, with 53% severe infections. Thirty-three of the 39 remitters received subsequent treatment consisting of maintenance chemotherapy courses in 17 patients, allogeneic stem cell transplantation (SCT) in 7 patients, and autologous SCT in 9 patients. The median overall survival of the entire cohort was 8.1 months, with 18% at 2.5 years. EMA chemotherapy using a single injection of mitoxantrone is effective in the treatment of high-risk AML. CR proportion was significantly higher in patients with a first CR duration > or =6 months when compared with those from a control trial using standard-dose mitoxantrone (90 vs 70%, p=0.03). PMID- 15782345 TI - Cardiac involvement by malignant lymphoma: a clinicopathologic study of 25 autopsy cases based on the WHO classification. AB - As cardiac involvement by malignant lymphoma (ML) is relatively uncommon and antemortem diagnosis is difficult, details of this condition remain to be elucidated. To clarify clinicopathologic features of cardiac lymphoma (CL), 25 autopsy cases were studied. Each was rediagnosed according to the World Health Organization (WHO) classification, and clinicopathologic characteristics were investigated by tumor phenotype. The study subjects were 13 males and 12 females with a mean age of 53.4 years. All cases were secondary CL and were not diagnosed as CL before death; 14 cases (56%) were of B-cell and 11 (44%) of T-cell (including natural killer cell) phenotype. Nasal and nasal-type natural killer/T cell lymphomas (NKTLs) accounted for five (20%). Cardiac manifestation was evident in eight (32%), with hematogenous infiltration as the most common pattern of tumor spread. Some B-cell CLs (n=3) were complicated by cardiac tamponade and heart failure (HF), and T-cell CLs (n=5), including three nasal NKTLs, also featured arrhythmia and sudden death. The incidence of T-cell phenotype was significantly elevated for CLs (p<0.05), especially for CLs with cardiac manifestation (p<0.01), compared with that for MLs in general. Our results indicate that T-cell lymphomas, compared with B-cell lymphomas, invade the heart more frequently and aggressively and are associated with a variety of cardiac manifestations. Where cardiac involvement is suspected, aggressive diagnostic procedures are warranted, especially with MLs having a T-cell phenotype. In cases of nasal NKTL, particular attention is necessary. PMID- 15782346 TI - Hydatid cyst of the renal pelvis. AB - Hydatid disease is fairly common and is prevalent worldwide. The principal organs involved are the liver and lungs. Kidney involvement also occurs but usually with involvement of other viscera. Isolated hydatid cyst of the kidney is extremely uncommon. In the kidney, the hydatid cyst usually occurs in the renal parenchyma. We present a rare occurrence of a hydatid cyst of the renal pelvis in a 4-year old child and discuss measures to decrease surgical morbidity. PMID- 15782347 TI - Photoinhibition of photosystem I. PMID- 15782348 TI - CaAlaAT1 catalyzes the alanine: 2-oxoglutarate aminotransferase reaction during the resistance response against Tobacco mosaic virus in hot pepper. AB - Hot pepper (Capsicum annuum L. cv. Bugang) plants exhibit a hypersensitive response (HR) upon infection by Tobacco mosaic virus (TMV) pathotype P0. To elucidate molecular mechanism that underlies this resistance, hot pepper cv. Bugang leaves were inoculated with TMV-P0 and genes specifically up-regulated during the HR were isolated by differential screening. One of the clones, CaAlaAT1 encoding a putative alanine aminotransferase (EC 2.6.1.2) exhibited organ-specific expression pattern and the transcript accumulated abundantly in red (ripe) fruit tissues. CaAlaAT1 transcript was also induced in older leaves during senescence. The expression of CaAlaAT1 gene was increased in the incompatible interaction with TMV-P0 but was not in the compatible interaction with TMV-P1.2. When a strain of Xanthomonas campestris pv. vesicatoria (Xcv) carrying an AvrBs2 gene was infiltrated into the leaves of a pepper cv. ECW 20R carrying Bs2 resistance gene, a marked induction and maintenance of CaAlaAT1 gene expression was observed. The expression of CaAlaAT1 gene was triggered by salicylic acid (SA) and ethylene but not by methyl jasmonate (MeJA). CaAlaAT1 seemed to be localized mostly at the cytosol from the polyethylene glycol (PEG) mediated transformation experiment. CaAlaAT1 seemed to catalyze alanine: 2 oxoglutarate aminotransferase (AKT) reaction, which was a main activity among the four activities in vitro, during the resistance response against TMV in hot pepper. These results suggest that CaAlaAT1, a protein known to be involved in metabolic reactions, might be one of the components in the plant's defense signal pathway against pathogens. PMID- 15782349 TI - Multiple frames of reference for pointing to a remembered target. AB - Pointing with an unseen hand to a visual target that disappears prior to movement requires maintaining a memory representation about the target location. The target location can be transformed either into a hand-centered frame of reference during target presentation and remembered under that form, or remembered in terms of retinal and extra-retinal cues and transformed into a body-centered frame of reference before movement initiation. The main goal of the present study was to investigate whether the target is stored in memory in an eye-centered frame, a hand-centered frame or in both frames of reference concomitantly. The task was to locate, memorize, and point to a target in a dark environment. Hand movement was not visible. During the recall delay, participants were asked to move their hand or their eyes in order to disrupt the memory representation of the target. Movement of the eyes during the recall delay was expected to disrupt an eye centered memory representation whereas movement of the hand was expected to disrupt a hand-centered memory representation by increasing movement variability to the target. Variability of movement amplitude and direction was examined. Results showed that participants were more variable on the directional component of the movement when required to move their hand during recall delay. On the contrary, moving the eyes caused an increase in variability only in the amplitude component of the pointing movement. Taken together, these results suggest that the direction of the movement is coded and remembered in a frame of reference linked to the arm, whereas the amplitude of the movement is remembered in an eye centered frame of reference. PMID- 15782350 TI - Effects of the Ebbinghaus figure on grasping are not only due to misjudged size. AB - It is not evident how the small effects of the flankers of the Ebbinghaus figure on peak grip aperture (PGA) should be interpreted. One interpretation is that the flankers influence the estimated size, which in turn influences the grasp. If this interpretation is correct, then only the size-dependent aspects of the grasping movement should depend on the spatial positions of the flankers. An alternative interpretation is that the effect on grip aperture is caused by a change in judgement of the required precision, in which case various aspects of the grasping movement could be influenced by the size and position of the flankers. We presented subjects with a display consisting of a central disk surrounded by four large or small flankers. The array of circular flankers could be rotated by 45 degrees . There were two tasks: to reproduce the perceived size of the central disk, and to grasp the central disk. As in other studies, the reproduced size and the PGA were both influenced by the size of the flankers. The effect on reproduced size settings was independent of the flankers' spatial position. Nevertheless, the flankers' position did influence the final grip aperture and the grip orientation at PGA and at movement offset. Because the flankers changed more than only the PGA, we conclude that the effect of the flankers on prehension cannot only be because of misjudgement of the size of the central disk. PMID- 15782354 TI - [Clinical utility of outpatient videoelectroencephalogram monitoring]. AB - OBJECTIVE: To analyze the utility of outpatient videoelectroencephalogram (VEEG) in a general neurology department to detect an ictal event. PATIENTS AND METHODS: One hundred and five patients with ictal phenomenology of unknown etiology, suspicion of pseudoseizures, refractory epilepsy with very frequent seizures, underwent outpatient VEEG monitoring from 30 minutes to five hours of duration, between June 1, 1999 and June 30, 2003. Patient medication was not modified to perform the recording. RESULTS: Among the 105 outpatient VEEG monitoring, 33 clinical pathologic events were identified; these comprised 14 epileptic seizures, 12 pseudoseizures, four syncopes, and three non epileptic abnormal movements. Outpatient VEEG monitoring duration was as follows: 30 minutes in 12 patients, between 30 minutes and two hours in another 12, and more than two hours in 9. In 19 patients, the VEEG recording allowed a definitive diagnosis; in one case, it changed the epileptic seizure type, and in 11 patients, it helped to better characterize the epileptic seizure type. CONCLUSION: Although the percentage of pathologic events during an outpatient VEEG monitoring of 30 minutes to five hours of duration is low, its clinical repercussion is very important and the added cost is low. PMID- 15782355 TI - [Spectral analysis of the electroencephalogram in children with systemic lupus erythematosus]. AB - INTRODUCTION: Systemic lupus erythematosus (SLE) is an autoimmune disease that affects different systems and organs, including the central nervous system (CNS). It has been suggested that about 40% of the cases of neuropsychiatric lupus (NPSLE) develop before SLE is diagnosed or at the same time it is being carried out, and 63% appear during the first year following diagnosis. AIMS: The aim of this study was to check the hypothesis that the electroencephalogram (EEG) may be sensitive to the damage to the CNS in children with SLE in whom there is still no clinical evidence of NPSLE. PATIENTS AND METHODS: EEG recordings were performed in 30 children with a diagnosis of SLE with or without signs of a neuropsychiatric syndrome. The results of the EEG were evaluated visually and analysed quantitatively. RESULTS: The visual inspection of the EEG showed the presence of alterations in 44.5% of the children with SLE and in 76.9% of those with NPSLE. There were significant differences in Student's t test (p = 0.0055) between the two groups for the analysis of the broadband spectral measurements. The narrow band analysis revealed a significant increase in the theta and delta frequencies in children with SLE as compared to standard values, whereas in children with NPSLE significant differences were found in the fast bands in frontal regions. CONCLUSIONS: Spectral analysis of the narrow band could help to confirm diagnoses of NPSLE, while anomalies in the slow bands could be an early marker of damage to the CNS although there are still no symptoms of the disease. PMID- 15782356 TI - [Non-valvular atrial fibrillation and completed stroke: factors determining mortality, recurrence and prognosis after a first event in the Mexican population]. AB - AIMS: Non-valvular atrial fibrillation (NVAF) accounts for 25% of completed strokes (CS) of a cardioembolic origin in patients over 60 years old. Our aim was to define the predictors of a good and poor prognosis after a CS secondary to an NVAF in our milieu. PATIENTS AND METHODS: We evaluated the risk factors (RF) and severity of CS in relation to death, functionality and recurrence at 5 years. 81 patients between the ages of 49 and 88 were followed up consecutively for 1 to 90 months; 38 (46.9%) of them were males. Multivariate analysis was performed with the following independent variables: age, gender, smoking, hypertension, heart disease, diabetes mellitus and characteristics of the stroke. The severity of the CS was assessed by means of the modified Rankin scale, which was dichotomised into a good prognosis (0-2) and a poor prognosis (> or = 3), both basal and at the end of the clinical control. We also evaluated the secondary preventive treatment used and its relation with recurrence, prognosis, death and complications. RESULTS: No RF was linked to a poor prognosis or recurrence; 88% had a poor prognosis. Antiplatelet drugs were used in 42% of cases and 39% received anticoagulants. A good final progression was observed in 9.5% of the patients treated with antiplatelet drugs versus 35% of those receiving anticoagulation therapy (p = 0.004). Severity of the CS on admission was worse in the aspirin group, with no differences in recurrence and mortality. A better prognosis was observed in patients from urban areas. CONCLUSIONS: Use of antiplatelet drugs, living in a rural area and a Rankin score of > or = 3 on admission are factors suggesting a poor prognosis in the clinical control at 5 years. PMID- 15782357 TI - [Thrombolysis in stroke: inappropriate consideration of the 'window period' as the time available]. AB - AIMS: The earlier r-TPA is administered in ischaemic strokes, the more effective it is. The aim of this study is to analyse the delay times in health care afforded in a consecutive series of cases that had received treatment, with a view to shortening them. PATIENTS AND METHODS: We analysed the medical records of the first patients to be treated in our centre. The paper describes several variables involving demographic and clinical factors, as well as the delay in entering the Emergency department, performing a CAT scan and especially the time elapsed between the CAT scan and starting treatment. We have examined the existence of an inappropriate correlation between delays that should be independent of one another. RESULTS: The mean age of the 17 patients treated was 68 years and they had a stroke severity score of 17 points on the NIHSS. The mean time of delay until arrival, arrival-CAT, and CAT-treatment were slightly under 1 hour each, and onset-treatment delay was 165 minutes, which is very close to the limit of the therapeutic window period. We found a strong inverse linear association between the time elapsed between onset and the CAT scan, and from the latter to the beginning of treatment (Spearman's r: -0.664, p = 0.004). CONCLUSIONS: Findings indicate that in our hospital, as in other centres in the initial phases of implementation, the therapeutic time window for intravenous thrombolysis in ischaemic stroke tends to run out. It must be highlighted that the resolve of the physician who indicates the treatment exerts a decisive effect on the delay. PMID- 15782358 TI - [Apraxia of gait: an acquired sequela with a poor prognosis]. AB - INTRODUCTION: Gait apraxia is not used to be considered as a diagnostic entity in Pediatric Neurology. CASE REPORTS: We present two pediatric patients that, after to have acquired normal gait and in consequence of a acute process, they lost the capacity to walk. In spite of intensive rehabilitation treatment hold along various years, they had not been able to help them. Both injury were very dissimilar; in one of them was affected the precentral and paracentral cortex in consequence of an encephalitic process. In the other, the basal ganglia and the hippocampus after a situation of near-drowning at the age of 15 months. CONCLUSION: The mechanism of this disorder is discussed and emphasis is done in its badly long-term prognosis. PMID- 15782359 TI - [The value of diffusion magnetic resonance imaging in diagnosing infarction of the anterior spinal artery]. AB - INTRODUCTION: The diagnosis of ischemic lesions affecting the spinal cord is sometimes difficult to confirm using conventional magnetic resonance imaging (MRI) techniques, and this makes it necessary to use those currently available to their full extent and also to search for new ones. CASE REPORT: We report the case of an 81-year-old male patient who presented symptoms of sudden onset paraparesis, secondary to lesions in the anterior spinal territory and whose diagnosis was reached using conventional spinal cord MRI, and later confirmed with the diffusion imaging technique. Diffusion MRI in the spinal cord territory, as well as in the brain territory, is a very interesting first choice technique when an ischemic pathology is suspected. Using only T2-weighted images does not allow acute ischemic lesions to be distinguished from chronic lesions; likewise, intramedullary T2 hyperintensity and standard DWI (Diffusion Weighted Imaging) is not specific to acute spinal ischemia, and the determination of the apparent diffusion coefficient in the lesion may help in the differential diagnosis of diseases affecting the spinal cord. CONCLUSIONS: Including diffusion RMI techniques in the study protocols in cases of ischemic spinal cord pathologies is indispensable today if we hope to reach a fast and correct diagnosis. PMID- 15782361 TI - [The psychobiology of learning and memory: fundamentals and recent advances]. AB - AIM: This review describes the concepts, temporal dynamics and main features of learning and memory systems from a comprehensive molecular, neuroanatomical, neurophysiological, cognitive and behavioural approach. DEVELOPMENT: Starting with molecular mechanisms of synaptic plasticity we describe the memory stages, implicit and explicit memory systems, working memory, remembering and forgetting. Each process is illustrated with examples of recent experimental and clinical research. CONCLUSIONS: Learning and memory are closely related brain processes which give rise to adaptive changes in behaviour. Implicit memory is a kind of unconscious and rigid memory for habits, which is based on brain regions processing perceptions and motor and emotional information, like the neocortex, the neostriatum, the cerebellum or the amygdala. Explicit or declarative memory is a conscious and flexible memory, hippocampus-dependent. Working memory is actually a system of executive cognition, based on interactions between the prefrontal cortex and other brain regions. The retrieval of complex memories consist of an active process of reconstruction of the past which incorporates new experiences of the subject who is remembering. The reactivation of memories can initiate genuine processes of reconsolidation and extinction. Forgetting could depend on alterations in the neural networks storing the information or, otherwise, on active processes which hinder consolidation or block the expression of the memories. PMID- 15782360 TI - [Improvements in the symptoms of olivopontocerebellar atrophy with gabapentin]. AB - INTRODUCTION: Olivopontocerebellar atrophy (OPCA) is a degenerative disease of the nervous system (NS) which currently has no known cure. The neuronal depopulation it brings about produces a number of neurochemical alterations, including a reduction in levels of gamma-aminobutyric acid (GABA) in tissues and in cerebrospinal fluid (CSF). The drug gabapentin (GBP) has proved to be capable of increasing the concentration of this neurotransmitter in the central nervous system, and of improving the cerebellar ataxia in other diseases with a similar neurochemical substrate. CASE REPORTS: We describe two sporadic cases of OPCA, who were administered GBP. In one of the cases, the ataxia was noticeably reduced after taking one 400 mg dose. In the other case, a considerable improvement was observed in a very intense cerebellar dysarthria, and there was less oscillopsia with better vision, following administration of GBP for a period of over 12 months. CONCLUSIONS: GBP has proved to be capable of slowing down the motor disorders reported by patients in the course of OPCA. We discuss how such effects are due to the increased levels of GABA in the NS triggered by the drug. Finally, we suggest that the administration of GBP could constitute an effective symptomatic treatment for the ataxia and the dysarthria caused by OPCA, and that the improvement in symptoms following single doses of GBP could be valuable in cases of OPCA, as well as other types of ataxia, that are ideal for taking advantage of the stimulus of the GABAergic neurotransmission. PMID- 15782362 TI - [Blepharospasm and blepharocolysis. Different sides of the same coin]. AB - INTRODUCTION: The eyelid movements are mediated mainly by the orbicularis oculi (OO) and the levator palpebrae superioris (LPS) muscles. Dissociated upper lid functions exhibit different counterbalanced action of these muscles, and in blinking they show a strictly reciprocal innervation. The disturbance of this close LPS-OO relationship likely leads to many of the central lid movement disorders. DEVELOPMENT: Two major lid postural disorders, the blepharospasm (BSP) and the blepharocolysis (BCO), share two different clinical aspects: the involuntary eyelids' closure along with the inability to open the eyes. BSP consists of an involuntary overactivity of the OO, with LPS co-contraction activity, and is expressed as frequent and prolonged blinks, clonic bursts, prolonged tonic contraction or a blend of all of them. BCO (commonly named 'so called lid opening apraxia') is an involuntary overinhibition of the LPS muscles with no evidence of ongoing OO activity; it exists a co-inhibition of these muscles. BSP and BCO occur in many instances of idiopathic dystonias and basal ganglia diseases and, less frequently, in rostral brainstem lesions. Both may coincide in the same patient. CONCLUSIONS: BSP and BCO should be considered different expressions of the dystonic focal eyelid movement disorders, related to the increase of the two counteracting components of the normal eyelid motor and blinking patterns: the excessive OO activation in the BSP; the excessive LPS inhibition in the BCO. PMID- 15782363 TI - [Guidelines for the clinical management of neuropathic pain (II)]. AB - INTRODUCTION: Up to 5% of the population suffers from neuropathic pain (NP). A bibliographical search in several databases revealed that, to date, there are no protocols to guide physicians who are not specialists in pain that enable them to treat NP and thus improve patients' quality of life. AIMS: The aim of this study is to provide Spanish-speaking physicians who are not specialists in pain with a set of guidelines for the treatment of NP. A bibliographical search was performed in order to base the results and conclusions on the evidence-based medicine methodology. DEVELOPMENT: First, we review the most effective clinical and paraclinical methods for diagnosing NP, and the LANSS pain scale is reported as the most appropriate method of clinically evaluating NP. The anatomical paths and the physiology of pain are then described and we review the molecular variables involved. Finally, we point out the current therapeutic options and propose an algorithm for the treatment of NP. CONCLUSIONS: There is no specific set of guidelines for the treatment of NP. At the present time, the keystone of NP treatment consists in the use of antidepressant and anticonvulsive drugs. There is a need for further clinical trials to prove the effectiveness of using combined medication. PMID- 15782364 TI - [Stroke in a young patient with the ascending pharyngeal artery mimicking dissection of carotid artery]. PMID- 15782365 TI - [Cerebral cryptococcosis in theoretically immunocompetent patients]. PMID- 15782366 TI - [Cryptococcosis in immunocompetent patients]. PMID- 15782368 TI - [Immunological disorders in epileptic patients are associated to the epileptogenic focus localization]. PMID- 15782369 TI - Design and size control of uniform zeolite nanocrystals synthesized in adjustable confined voids formed by recyclable monodisperse polymer spheres. PMID- 15782370 TI - Synthesis and characterization of the nitrogen-centered trigonal prismatic clusters [W6NCl18](n-) (n=1-3). PMID- 15782371 TI - Kinetics of photoinduced RNA refolding by real-time NMR spectroscopy. PMID- 15782372 TI - Hydroxylation of benzene to phenol under air and carbon monoxide catalyzed by molybdovanadophosphoric acid. PMID- 15782373 TI - Stable, water-soluble Pta-based Ru-Ag organometallic polymers. PMID- 15782374 TI - Alkenylcarbene ruthenium arene complexes as initiators of alkene metathesis: an enyne creates a catalyst that promotes its selective transformation. PMID- 15782375 TI - An electron-deficient iridium(III) dihydride complex capable of intramolecular C- H activation. PMID- 15782376 TI - A ligand composed of dinitrogen and methyldiphenylphosphane in a cationic molybdenum complex. PMID- 15782377 TI - High-affinity multivalent DNA binding by using low-molecular-weight dendrons. PMID- 15782378 TI - Metal-rich, neutral and cationic organotin clusters. PMID- 15782382 TI - Borane mimics of C1-Mm organometallic complexes. PMID- 15782383 TI - C10-modified artemisinin derivatives: efficient heme-alkylating agents. PMID- 15782385 TI - Actinyl peroxide nanospheres. PMID- 15782388 TI - Association of human endogenous retroviruses with multiple sclerosis and possible interactions with herpes viruses. AB - The hypothesis that human endogenous retroviruses (HERVs) play a role in autoimmune diseases is subject to increasing attention. HERVs represent both putative susceptibility genes and putative pathogenic viruses in the immune mediated neurological disease multiple sclerosis (MS). Gammaretroviral HERV sequences are found in reverse transcriptase-positive virions produced by cultured mononuclear cells from MS patients, and they have been isolated from MS samples of plasma, serum and CSF, and characterised to some extent at the nucleotide, protein/enzyme, virion and immunogenic level. Two types of sequences, HERV-H and HERV-W, have been reported. No known HERV-H or HERV-W copy contains complete ORFs in all prerequisite genes, although several copies have coding potential, and several such sequences are specifically activated in MS, apparently resulting in the production of complete, competent virions. Increased antibody reactivity to specific Gammaretroviral HERV epitopes is found in MS serum and CSF, and cell-mediated immune responses have also been reported. Further, HERV-encoded proteins can have neuropathogenic effects. The activating factor(s) in the process resulting in protein or virion production may be members of the Herpesviridae. Several herpes viruses, such as HSV-1, VZV, EBV and HHV-6, have been associated with MS pathogenesis, and retroviruses and herpes viruses have complex interactions. The current understanding of HERVs, and specifically the investigations of HERV activation and expression in MS are the major subjects of this review, which also proposes to synergise the herpes and HERV findings, and presents several possible pathogenic mechanisms for HERVs in MS. PMID- 15782389 TI - Hepatitis C virus and the immune system: a concise review. AB - Hepatitis C Virus (HCV) induces a chronic infection in 50%-80% of infected individuals, which can lead to cirrhosis and hepatocellular carcinoma. The inefficiency of the immune system in eliminating the virus is not well understood as humoral and cellular immune responses are induced. While a persistent infection is generally associated with a weak CD4+ and CD8+ T cell response during the acute phase, there is no good explanation as to why this response is strong enough in 20% of acutely infected people such that they spontaneously resolve the infection. However, the immune system partially controls the viral infection but due to a long-lasting inflammatory milieu, hepatic damage occurs. During the chronic phase of the infection, HCV does not seem to be cytopathic. This aspect is still controversial as the virus was linked to the development of cholestatic syndrome or acute lobular hepatitis after liver transplant in HCV infected patients. The development of new experimental systems such as HCV pseudoparticles, genomic replicon and transfected cell lines have improved our vision of the virus cycle as well as the understanding of the mechanism of persistence. However, a convincing explanation for the chronicity of the infection in the presence of a functional immune response is still missing and is an important area of research to understand HCV immune pathogenesis. Future research should dissect mechanisms that lead to quantitatively or qualitatively inadequate immune responses, the role of the high variability of the virus, the relevance of host's genetic factors and mechanisms of immunosuppression induced by the virus. PMID- 15782390 TI - Randomized trial comparing the Prolene Hernia System, mesh plug repair and Lichtenstein method for open inguinal hernia repair (Br J Surg 2005; 92: 33-38). PMID- 15782391 TI - Randomized clinical trial of open versus laparoscopic cholecystectomy in the treatment of acute cholecystitis (Br J Surg 2005; 92: 44-49). PMID- 15782392 TI - Fast-track surgery (Br J Surg 2005; 92: 3-4). PMID- 15782394 TI - Combination of liquid chromatography/tandem mass spectrometry and gas chromatography/mass spectrometry for the detection of 21 anabolic steroid residues in bovine urine. AB - For the detection of anabolic steroid residues in bovine urine, a highly sensitive liquid chromatographic/electrospray ionization tandem mass spectrometric (LC/ESI-MS/MS) method was developed using both positive and negative ionization. For four compounds the ESI mode was not sensitive enough and gas chromatographic/mass spectrometric GC/MS detection was therefore still necessary as a complementary method. The sample clean-up consisted of solid-phase extraction (SPE) on a C(18) column followed by enzymatic hydrolysis and a second solid-phase extraction on a combination of a C(18) and a NH(2) column. After this last SPE clean-up, the eluate was split into two equal aliquots. One aliquot was further purified and after derivatization used for GC/MS analysis. The other aliquot was analyzed with LC/MS/MS in both ESI+ and ESI- modes. The method was validated according to the European Commission Decision 2002/657/EC. Decision limits (CCalpha) were between 0.16 and 1 ng ml(-1) for the compounds detected with the LC/MS/MS method. The developed method is used in routine analysis in our laboratory. PMID- 15782395 TI - Psychosocial impacts of a camping experience for children with cancer and their siblings. AB - BACKGROUND: We conducted a prospective two-group evaluation of pediatric cancer patients and their siblings regarding experiences and affective changes resulting from a 1-week summer camp experience. METHODS: The patients and siblings were assessed prior to camp (Baseline), at the end of camp (Follow-up 1), and again 4 6 months later (Follow-up 2). Assessments included standardized tests for depressive affects, social competency, and a measure of pleasure and participation in camp activities. RESULTS: Sixty-six children were assessed, including 31 (47%) patients and 35 (53%) siblings. Ages ranged from 7 to 17 years. Of the patient campers 19 (61%) had leukemia or lymphoma and 12 (39%) had solid tumors. Results showed marked changes in affective symptoms for patient campers over time (improvements), not shown by sibling campers. For patient campers these affective changes were not present immediately after camp, but were quite significant when measured 4-6 months later. Both patient and sibling campers reflected the same positive memories and pleasure in camp activities over time. For neither group did memories or pleasure fade over time. The camping experience did not have differential impacts on first time versus returning campers. Twelve campers (18% of sample) indicated suicidal ideation on the measure of depressive affects. They did well at camp and presented no special management issues. CONCLUSION: Expectations appear substantially different for patient versus sibling campers. The camping experience appeared to impact these groups differently, with patient campers impacted in ways not experienced by sibling campers. PMID- 15782396 TI - High-resolution crystallography and drug design. AB - Ultra-high-resolution X-ray crystallography of macromolecules (i.e. resolution better than 0.8 Angstroms) is a rising field that promises to provide new insight into the structure-function relationships of biomacromolecules. The picture emerging from macromolecular structures at this resolution is far more complex than previously understood, requiring for its study improved tools for structure refinement, analysis and annotation. Some of these problems were highlighted during the recent High Resolution Drug Design Meeting (Bischenberg-Strasbourg, France, 13-16 May 2004). We will review here some of the results and discussions that took place during that meeting and elaborate on the trends and challenges ahead in this emerging new field of research. PMID- 15782397 TI - The history of disproportionality measures (reporting odds ratio, proportional reporting rates) in spontaneous reporting of adverse drug reactions. PMID- 15782398 TI - Statin-associated rhabdomyolysis. PMID- 15782400 TI - A DFT study on hetero-Diels-Alder reactions catalyzed by cobalt complexes: Lewis acidity enhancement as a consequence of spin transition caused by Lewis base coordination. PMID- 15782399 TI - Gly482Ser polymorphism of the peroxisome proliferator-activated receptor-gamma coactivator-1 gene might be a risk factor for diabetic retinopathy in Slovene population (Caucasians) with type 2 diabetes and the Pro12Ala polymorphism of the PPARgamma gene is not. AB - BACKGROUND: The peroxisome proliferator-activated receptor-gamma (PPARgamma) gene has been recently associated with type 2 diabetes, obesity and traits depending on VEGF expression (e.g. retinopathy). The PPARgamma gene and its coactivator, the peroxisome proliferator-activated receptor-gamma coactivator-1 (PPARGC1) gene, have been implicated to be involved in glucose uptake and altered lipid oxidation. We therefore hypothesized that the Gly482Ser polymorphism of the PPARGC1 gene and Pro12Ala polymorphism of the PPARgamma gene might confer susceptibility to diabetic retinopathy in type 2 diabetes. The aim of this study was to investigate the association between the Pro12Ala polymorphism in the PPARgamma gene and Gly482Ser polymorphism in the PPARGC1 gene and the development of diabetic retinopathy in the Slovene population (Caucasians) with type 2 diabetes. METHODS: One hundred and sixty subjects with type 2 diabetes and diabetic retinopathy were compared with 101 diabetic subjects without diabetic retinopathy. Chi-square test was used to compare discrete variables, and continuous clinical data were compared by unpaired students t - test. RESULTS: A significantly higher frequency of the AA genotype of the Gly482Ser polymorphism of the PPARGC1 gene was found in the patients with diabetic retinopathy compared to the patients without diabetic retinopathy (14.4% vs 5.9%; p = 0.035), whereas the Pro12Ala polymorphism of the PPARgamma gene failed to yield an association with diabetic retinopathy. CONCLUSIONS: The present study demonstrates that the AA genotype of the Gly482Ser polymorphism in the PPARGC1 gene might be a risk factor for diabetic retinopathy in the Slovene population (Caucasians) with type 2 diabetes (odds ratio 2.7, 95% confidence interval 1.0-6.8), whereas the Pro12Ala polymorphism of the PPARgamma gene failed to confer susceptibility to diabetic retinopathy. PMID- 15782401 TI - Phenolate complexes of iron(II) in different spin states. PMID- 15782403 TI - Reduced intensity hematopoietic stem-cell transplantation across human leukocyte antigen barriers in a patient with congenital amegakaryocytic thrombocytopenia and monosomy 7. AB - Congenital amegakaryocytic thrombocytopenia (CAMT) is a rare inherited bone marrow failure syndrome that has the potential to progress to pancytopenia and acute myeloid leukemia. Hematopoietic stem-cell transplantation (HSCT) is presently the only curative treatment approach. We used a reduced intensity transplantation regimen in a CAMT patient with aplastic anemia and monosomy 7 who had no matched related donor. The patient had rapid and durable engraftment with minimal complications and is well 24 months post-transplantation. Thus, reduced intensity conditioning might be a feasible approach to stem-cell transplantation in patients with CAMT who do not have a related donor and who are at increased risk of toxicity from standard conditioning regimens. PMID- 15782402 TI - Prognostic factors for relapsed childhood acute lymphoblastic leukemia: impact of allogeneic stem cell transplantation--a report from the Kyushu-Yamaguchi Children's Cancer Study Group. AB - BACKGROUND: The treatment results of childhood acute lymphoblastic leukemia (ALL) with a first relapse were retrospectively analyzed to determine prognostic factors. In particular, an attempt was made to clarify whether stem cell transplantation (SCT) had any advantages over chemotherapy. PROCEDURES: Of the 407 children with ALL diagnosed between 1984 and 1996, 117 suffered from a relapse before December 1999. The patients were treated differently according to the protocols of each institution. The potential prognostic factors examined were: the time of initial diagnosis, gender, immunophenotype of leukemic blasts and the NCI-risk classification at initial diagnosis, the site of relapse, the time of relapse (early: within 18 months after diagnosis, intermediate: other than either early or late relapse, late: later than 6 months after the discontinuation of front-line chemotherapy), and the treatment after relapse (chemotherapy alone and SCT). RESULTS: A second complete remission (CR2) was achieved in 90 patients (77%) and thirty of them maintained CR2, thus resulting in an event-free survival rate (EFS) of 25.1% and an overall survival rate of 26.1%. The significant prognostic factors identified by a multivariate analysis included the time of relapse (EFS: early 16.2%, intermediate 23.9%, late 35.1%, P = 0.012) and the treatment after relapse (EFS: SCT 30.3%, chemotherapy 22.0%, P = 0.049). When patients with an isolated bone marrow relapse and continuous CR2 for more than 3 months were analyzed, the treatment in CR2 was the only independent prognostic factor (EFS: SCT 60.2%, chemotherapy 25.7%, P = 0.005). CONCLUSIONS: In children with ALL and a first relapse, the time of relapse and the treatment after relapse were found to be independent prognostic factors. Allogeneic SCT in CR2 showed significantly better results than chemotherapy in patients with an isolated bone marrow relapse. PMID- 15782404 TI - Effectiveness of chemotherapy in non-rhabdomyosarcoma soft tissue sarcomas. PMID- 15782405 TI - Highly efficient strategy for enhancing taxoid production by repeated elicitation with a newly synthesized jasmonate in fed-batch cultivation of Taxus chinensis cells. AB - A highly efficient bioprocessing strategy was developed for enhancing the production of plant secondary metabolites by repeatedly eliciting a fed-batch culture with a newly synthesized powerful jasmonate analog, 2,3-dihydroxypropyl jasmonate (DHPJA). In suspension cultures of a high taxuyunnanine C (Tc) producing cell line of Taxus chinensis, 100 microM DHPJA was added on day 7 to fed-batch cultures with feeding of 20 g L(-1) sucrose on the same day. The synergistic effect of elicitation and substrate feeding on Tc biosynthesis was observed, which resulted in higher Tc accumulation than that by elicitation or sucrose feeding alone. More interestingly, both specific Tc yield (i.e., Tc content) and volumetric yield was further improved by a second addition of 100 microM DHPJA (on day 12) to the fed-batch cultures. In particular, with repeated elicitation and sucrose feeding the Tc volumetric yield was increased to 827 +/- 29 mg L(-1), which was 5.4-fold higher than that of the nonelicited batch culture. Furthermore, the above novel strategy was successfully applied from shake flask to a 1-L airlift bioreactor. A high Tc production and productivity of 738 +/- 41 mg L(-1) and 33.2 +/- 1.9 mg L(-1) d(-1), respectively, was achieved, which is higher than previous reports on Tc production in bioreactors. The results suggest that the aforementioned bioprocessing strategy may potentially be applied to other cell culture systems for efficient production of plant secondary metabolites. PMID- 15782406 TI - Catheter-related bacteremia due to Roseomonas species in pediatric hematology/oncology patients. AB - Roseomonas is a newly described genus of pink-pigmented, gram-negative bacteria. Human infections caused by Roseomonas species are very rare. We report two cases of central venous catheter-related bacteremia associated with Roseomonas species (one case with R. gilardii and one with R. fauriae), and review the clinical spectrum of previously reported cases in the literature. Clinicals should be aware that Roseomonas species may cause serious infections in children. PMID- 15782407 TI - Evaluation of transcobalamin II polymorphisms as neural tube defect risk factors in an Irish population. AB - BACKGROUND: Decreased maternal folate levels are associated with having a child with a neural tube defect (NTD), and periconceptual folic acid supplementation reduces this risk by >50%. Vitamin B(12) (as methylcobalamin) is a cofactor for methionine synthase, an enzyme that plays a key role in folate metabolism. Alterations in vitamin B(12) metabolism may influence the development of NTDs. Low levels of maternal plasma vitamin B(12) and reduced binding of vitamin B(12) by transcobalamin II (TCII) are independent risk factors for NTDs. TCII levels are altered in the amniotic fluid of pregnancies affected by NTDs. Given this evidence, inherited variants in genes involved in vitamin B(12) trafficking such as TCII are candidate NTD risk factors. METHODS: We used case/control and family based association methods to investigate whether six common polymorphisms in the TCII gene influence NTD risk. TCII genotypes were determined for more than 300 Irish NTD families and a comparable number of Irish controls. RESULTS: Allele and genotype frequencies for each polymorphism did not differ between family members and controls. CONCLUSIONS: These six TCII polymorphisms do not strongly influence NTD risk in the Irish population. The Supplementary Material for this article can be found on the Birth Defects Research (Part A) website: http://www.mrw.interscience.wiley.com/suppmat/1542 0752/suppmat/2005/73/v73.4.swanson.html PMID- 15782408 TI - Electrochemical evaluation of cellular physiological status under stress in Escherichia coli with the rpoS-lacZ reporter gene. AB - We developed an electrochemical detection method for evaluating cellular physiological status based on the stringent response as a means to monitor cell viability. A reporter plasmid was constructed by inserting the beta-galactosidase gene (lacZ) under the control of the rpoS promoter, and then used to transform E. coli cells. Electrochemical responses from the products catalyzed by beta galactosidase expressed by these E. coli cells were detected using the chronoamperometric technique in a nondestructive manner. Comparisons of response currents between the relA-positive strain and relA-negative strain revealed that increases in these currents were caused by the stringent response due to the stressful alcoholic environment, and thus as a model of stressful cultivating conditions. The current was proportional to the beta-galactosidase activity assayed by a conventional method that required the destruction of cells. The cellular physiological status, which depends on the stringent response as a viability marker, therefore, could then be evaluated online with a current using the rpoS-lacZ reporter gene in the relA-positive strain without pretreatment. PMID- 15782410 TI - Renpenning syndrome comes into focus. AB - Renpenning syndrome represents a prototypic X-linked mental retardation condition with full expression of the phenotype in males and little or no expression in females. The predominant clinical findings are microcephaly, long narrow face, short stature with lean body build, and small testes. Mental retardation, usually of severe degree, occurs in 95% of cases. Less than 20% of cases have major malformations, the most common being cardiac defects and cleft palate. Subsequent to the description of mutations in the polyglutamine tract binding protein 1 (PQBP1) in Sutherland-Haan syndrome, Hamel cerebropalatocardiac syndrome, MRX55, and two small XLMR families, a single nucleotide insertion has been found in the original family with Renpenning syndrome and an AGAG deletion in a second family with the Renpenning syndrome. Mutations have also been found in Golabi-Ito-Hall syndrome, Porteous syndrome, and an additional small family. It is now demonstrated that five named XLMR syndromes (Sutherland-Haan, Hamel cerebropalatocardiac, Golabi-Ito-Hall, Porteous, and Renpenning), one nonsyndromic family (MRX55), and three small XLMR families have PQBP1 mutations and are thus allelic XLMR entities. In acknowledgement of the historical importance of the original report of Renpenning syndrome [1962], we propose that the entities with PQBP1 mutations be combined under the name of Renpenning syndrome. PMID- 15782412 TI - Cardiovascular genetics clinics. PMID- 15782413 TI - Spatial and temporal expression of S100B in cells of oligodendrocyte lineage. AB - The analysis of oligodendrocyte (OL) lineage development has been facilitated by the immunocytochemical characterization of OL-specific antigens and definition of the phenotypes sequentially acquired by differentiating OLs. The purpose of the present study was to address an enduring discrepancy between several reported cases of S100B immunodetection in CNS myelin and myelinating OLs on the one hand, and the systematic use of the S100B protein as an alleged astrocytic marker in studies of the mammalian CNS on the other. To resolve this discrepancy, we have compared the developmental distribution of EGFP+ cells in the CNS of s100b enhanced green fluorescent protein (EGFP) (Vives et al., 2003) and cnp-EGFP (Yuan et al., 2002) mice, and examined the degree of overlap between EGFP expression and that of stage-specific markers of OL differentiation during the embryonic and postnatal phases of development. We demonstrate that the S100B protein is expressed in postnatal and adult populations of NG2+ progenitors of mouse brain, as well as in immature and mature myelinating OLs present in the brain and spinal cord of embryonic and adult mice, respectively. Comparison between EGFP and endogenous S100B expression in the s100b-EGFP and cnp-EGFP mice indicates that S100B protein expression is upregulated in immature and mature OLs. These results argue against the current view that S100B expression is restricted to the astrocytic lineage in the CNS, and indicate that the use of S100B in combination with other molecular markers will help discriminate oligodendrocytes from astrocytes. PMID- 15782414 TI - Practical population group assignment with selected informative markers: characteristics and properties of Bayesian clustering via STRUCTURE. AB - Population stratification, which is caused by population genetic substructure (PGS), is a critical issue for the design and interpretation of genetic association studies. Methods to address this problem have been devised, but little is known at this point about practical genotyping requirements for resolving PGS based on different marker characteristics. In this report, we seek to (1) identify a small, practical marker set to differentiate African Americans (AAs) from European Americans (EAs), and (2) assess the impact of marker efficiency and sample size on clustering individuals into subgroups by the methods of STRUCTURE (Pritchard et al., [2000a] Genetics 155:945-959). A panel of 37 markers was genotyped for 865 individuals (640 EAs and 225 AAs) from the Northeastern United States. Among EAs, the assignment accuracy reached >99% using only the 4 most efficient markers. Among AAs, the assignment accuracy exceeded 95% when using the 6 most informative markers. Smaller sample size increased the variance in population differentiation, rather than degrading the results consistently. We conclude that the use of marker-efficiency measures for marker selection yielded a relatively small set of STR markers that were effective at differentiating EA and AA populations. The number of markers required is much lower than has been suggested in previous studies. PMID- 15782415 TI - Premature death of adult adoptees: analyses of a case-cohort sample. AB - Genetic and environmental influence on risk of premature death in adulthood was investigated by estimating the associations in total and cause-specific mortality of adult Danish adoptees and their biological and adoptive parents. Among all 14,425 non-familial adoptions formally granted in Denmark during the period 1924 through 1947, we selected the study population according to a case-cohort sampling design. As the case-control design, the case-cohort design has the advantage of economic data collection and little loss in statistical efficiency, but the case-cohort sample has the additional advantages that rate ratio estimates may be obtained, and re-use of the cohort sample in future studies of other outcomes is possible. Analyses were performed using Kalbfleisch and Lawless's estimator for hazard ratio, and robust estimation for variances. In the main analyses the sample was restricted to birth years of the adoptees 1924 and after, and age of transfer to the adoptive parents before 7 years, and age at death was restricted to 16 to 70 years. The results showed a higher mortality among adoptees, whose biological parents died in the age range of 16 to 70 years; this was significant for deaths from natural causes, vascular causes and all causes. No influence was seen from early death of adoptive parents, regardless of cause of death. PMID- 15782416 TI - Multipotent stem cells from adult olfactory mucosa. AB - Multipotent stem cells are thought to be responsible for the generation of new neurons in the adult brain. Neurogenesis also occurs in an accessible part of the nervous system, the olfactory mucosa. We show here that cells from human olfactory mucosa generate neurospheres that are multipotent in vitro and when transplanted into the chicken embryo. Cloned neurosphere cells show this multipotency. Multipotency was evident without prior culture in vitro: cells dissociated from adult rat olfactory mucosa generate leukocytes when transplanted into bone marrow-irradiated hosts, and cells dissociated from adult mouse olfactory epithelium generated numerous cell types when transplanted into the chicken embryo. It is unlikely that these results can be attributed to hematopoietic precursor contamination or cell fusion. These results demonstrate the existence of a multipotent stem-like cell in the olfactory mucosa useful for autologous transplantation therapies and for cellular studies of disease. PMID- 15782417 TI - Significance of bifid epiglottis. AB - Bifid epiglottis is a rare anomaly, which is heterogeneous and is often associated with other anomalies, particularly polydactyly. It has been reported in 40% of patients with Pallister-Hall syndrome and rarely in other syndromes. We report two brothers with bifid epiglottis who also have features suggestive of Bardet-Biedl syndrome. We also review the features seen in 22 patients reported in the literature with bifid epiglottis. No patient had bifid epiglottis as an isolated anomaly. Other malformations include clefts, micropenis, renal abnormalities, anal malformations, hypospadias, hypothalamic hamartomas, hypopituitarism, heart defects, and Hirschprung disease. Bifid epiglottis may be an under-recognized feature of Bardet-Biedl syndrome and should be considered in these patients, particularly if there are airway symptoms. Many of the anomalies associated with bifid epiglottis have potentially serious consequences and thus, a thorough evaluation of the patient with bifid epiglottis is warranted. PMID- 15782418 TI - New anticoagulants: a pediatric perspective. AB - Unfractionated heparin and vitamin K antagonists such as warfarin have been used as the anticoagulants of choice for over five decades. Subsequently, low molecular weight heparins (LMWHs) became widely available and have provided several advantages, especially in infants and children. The field of anticoagulation, however, has undergone a major revolution with better understanding of the structure of coagulation proteins and the development of a host of new drugs with highly specific actions. Many of these drugs have undergone extensive clinical testing in adults and have been approved for specific indications in adults. Unfortunately, clinical data and the reported use of these drugs in children are extremely limited. A lack of familiarity with the actions and pharmacokinetic properties of these drugs could be a major contributing factor. This review focuses on several of the new anticoagulants, with a special emphasis on those that could be potentially beneficial in pediatric patients with thromboembolic disorders. The need for well-designed trials with large-scale participation by pediatric hematologists in order to improve the antithrombotic care of young infants and children is also emphasized. PMID- 15782419 TI - Hemiparkinsonism and levodopa-induced dyskinesias after focal nigral lesion. AB - We present a patient with tremor-dominant hemiparkinsonism after a focal lesion to the substantia nigra. An excellent response to levodopa was complicated by rapid development of motor fluctuations and disabling dyskinesias. Stereotactic thalamotomy resulted in a persistent extinction of parkinsonism and of dyskinesias along with stopping dopaminergic treatment. PMID- 15782420 TI - Progressive supranuclear palsy-like phenotype associated with bilateral hypoxic ischemic striopallidal lesions. AB - A progressive supranuclear palsy (PSP)-like syndrome due to vascular or anoxic brain insult is rare. We describe a 65-year-old man with a progressive PSP-like phenotype associated with hypoxic-ischemic bilateral striopallidal lesions, secondary to rupture of and subsequent surgery for a thoracic aortic aneurysm. After early extrapyramidal features, 10 months later he started to fall, and developed levator inhibition. A supranuclear gaze palsy for downgaze was documented 5.5 years after the insult. PMID- 15782421 TI - Depressive symptoms and life satisfaction in patients with multiple system atrophy. AB - Multiple system atrophy (MSA) is a progressive neurodegenerative disorder characterized by extrapyramidal signs, prominent autonomic failure, and a poor prognosis. In the absence of restorative treatment, management is aimed at improving quality of life. Little is known about modifiable factors, such as depression, that may affect quality of life in MSA. The present study investigated the rate of depressive symptoms and their relationship to life satisfaction in patients with MSA. Ninety-nine patients with MSA (54% women; mean age, 67.8 +/- 8.8) completed measures of depressive symptoms, life satisfaction, physical function, and disease and demographic factors. Objective autonomic indices were abstracted from the medical chart. Participants reported a high rate of depressive symptoms, with 39% endorsing moderate to severe depressive symptoms on the Beck Depression Inventory (BDI > or = 7). Reported life satisfaction was low, with a mean of 38.8 on a 100-point visual analogue scale (0 = Extremely Dissatisfied, 100 = Extremely Satisfied). The SF-36 Physical Component Scale was approximately 1.5 standard deviations below the mean of a normative sample of healthy adults the same age. Regression analysis revealed that autonomic disease parameters accounted for 22% of the variance in life satisfaction. Physical function did not account for any additional variance; however, depressive symptoms accounted for an additional 15%. Depressive symptoms are common, often severe, and an important determinant of life satisfaction in patients with MSA. Adequate treatment of comorbid depression may improve quality of life in this population, despite the presence of other debilitating deficits. PMID- 15782422 TI - Quality of life, mucositis, and xerostomia from radiotherapy for head and neck cancers: a report from the NCIC CTG HN2 randomized trial of an antimicrobial lozenge to prevent mucositis. AB - BACKGROUND: The National Cancer Institute of Canada Clinical Trials Group undertook a multicenter, randomized, double-blind controlled trial of an oral antimicrobial versus placebo to prevent and treat mucositis. We present the quality of life (QOL) analysis for this trial. METHODS: One hundred thirty-eight patients were randomly assigned. QOL data were collected every 2 weeks before, during, and after radiotherapy. The European Organization for Research and Treatment of Cancer Quality of Life questionnaire (EORTC QLQ-C30) and a Trial Specific Checklist (TSC) were used. RESULTS: The antimicrobial lozenge did not impact QOL. The principal acute side effect of radiotherapy is oral pain, affecting more than 90% of patients. Role function is impacted during treatment, and patients experience fatigue. Appetite was reported to markedly increase during radiotherapy. There was a dramatic and persistent increase in dry mouth. CONCLUSIONS: This study highlights the benefits of combining the EORTC QLQ-30 with an "oral" TSC in a randomized controlled trial and provides valuable baseline data for their use with an objective mucositis scoring system. PMID- 15782423 TI - An interaction between apolipoprotein E and TERE1 with a possible association with bladder tumor formation. AB - TERE1, a recently discovered gene/protein appears to play a role in bladder tumor growth regulation but to date does not have clear functional correlates. The objective of this study was to gain further insight into the function of the TERE1 protein by identifying potential protein to protein interactions with TERE1 and determining whether these interactions are associated with putative growth regulatory pathways and/or bladder tumor formation. Towards this aim, we have performed a bacterial two hybrid assay and isolated interacting clones, which then were sequenced and further examined by affinity chromatography and immunoprecipitation. From among several positive clones, we isolated a putative interacting plasmid containing the C-terminal portion of preapolipoprotein E starting from amino acid number 124 from the pBT-TERE1/pTarget-cDNA bacterial two hybrid system. The C-terminal portion of apoE interaction with the TERE1 was confirmed using ProBond columns by the expression of 6XHis recombinant and (35)S methionine/cysteine labeled proteins. We found that there was ubiquitous expression of the apoE transcript in normal bladder and in various grades and stages of transitional cell carcinoma (TCC) of the bladder. Likewise, we detected the apoE protein in both normal and malignant bladder tissues by Western blot. There was a significant decrease in the apoE protein in 12 of 16 muscle invasive TCCs of the bladder compared to normal bladder mucosa samples. Previous studies in rat fibroblasts have found that expression of apoE can decrease the phosphorylation of the growth factor-related p42/44 MAP kinase. A significant decrease in p44/p42 MAPK phophorylation was also apparent using a phosphorylation specific antibody in human 293 kidney cells upon transfection and expression of apoE. In conclusion, the results from this study suggest that the expression and regulation of the apoE pathway may yield clues toward understanding the function of TERE1. PMID- 15782424 TI - Initial symptoms and reactions to trauma-related stimuli and the development of posttraumatic stress disorder. AB - We investigated laboratory and experimental variables as predictors of the development of posttraumatic stress disorder (PTSD). Evoked heart rate response to trauma-related pictures, attentional bias in the dot-probe task, and viewing time were assessed in 35 victims of a traumatic event and again after 3 months. Data was compared to 26 control participants. At first assessment trauma victims showed heart rate (HR) acceleration and controls showed HR deceleration to trauma related material. The group of trauma victims improved clinically over time. Predictors of the number of PTSD symptoms after 3 months were re-experiencing (33% of the variance) and amplitude of the evoked HR reaction to trauma-related pictures (15%). The two variables were highly correlated. Trauma victims were also more anxious, viewed trauma-related pictures for a longer time, and had a longer reaction time in the dot-probe task (but no distinct attentional bias) than control participants. Results indicate that specific fear responses and re experiencing contribute to the development of posttraumatic stress disorder. PMID- 15782425 TI - Behavioral and cardiovascular effects of 7.5% CO2 in human volunteers. AB - The study of carbon dioxide (CO2) inhalation in psychiatry has a long and varied history, with recent interest in using inhaled CO2 as an experimental tool to explore the neurobiology and treatment of panic disorder. As a consequence, many studies have examined the panic-like response to the gas either using the single or double breath 35% CO2 inhalation or 5-7% CO2 inhaled for 15-20 min, or rebreathing 5% CO2 for a shorter time. However, this lower dose regime produces little physiological or psychological effects in normal volunteers. For this reason we have studied the effects of a higher concentration of CO2, 7.5%, given over 20 min. Twenty healthy volunteers were recruited to a double blind, placebo controlled study where air and 7.5% CO2 were inhaled for 20 min. Cardiovascular measures and subjective ratings were obtained. When compared to air, inhaling 7.5% CO2 for 20 min increases systolic blood pressure and heart rate, indicating increased autonomic arousal. It also increases ratings of anxiety and fear and other subjective symptoms associated with an anxiety state. The inhalation of 7.5% CO2 for 20 min is safe for use in healthy volunteers and produces robust subjective and objective effects. It seems promising as an anxiety provocation test that could be beneficial in the study of the effects of anxiety on sustained performance, the discovery of novel anxiolytic agents, and the study of brain circuits and mechanisms of anxiety. PMID- 15782426 TI - Studies of protein folding and structure with model peptides. PMID- 15782427 TI - From oligopeptides to sweet proteins. PMID- 15782428 TI - Isosteric replacement of sulfur with other chalcogens in peptides and proteins. AB - The review addresses the functional and structural properties of the two series of chalcogen analogues of amino acids in peptides and proteins, the methionine and the serine/cysteine series, and discusses the synthesis of the related selenium/tellurium analogues as well as their use in peptide synthesis and protein expression. Advances in synthetic methodologies and recombinant technologies and their combined applications in native and expressed protein ligation allows the isomorphous character of selenium- and tellurium-containing amino acids to be exploited for production of heavy metal mutants of proteins and thus to facilitate the phasing problem in x-ray crystallography. In addition, selenocysteine has been recognized as an ideal tool for the production of selenoenzymes with new catalytic activities. Moreover, the fully isomorphous character of disulfide replacement with diselenide is well suited to increase the robustness of cystine frameworks in cystine-rich peptides and proteins and for the de novo design of even non-native cystine frameworks by exploiting the highly negative redox potential of selenols. PMID- 15782429 TI - Conformational investigation of alpha,beta-dehydropeptides. XV: N-acetyl alpha,beta-dehydroamino acid N 'N '-dimethylamides: conformational properties from infrared and theoretical studies. AB - The FTIR spectra were analysed in the region of the nu(s)(N-H), AI(C=O) and nu(s)(Calpha=Cbeta) bands for a series of Ac-DeltaXaa-NMe2, where DeltaXaa = DeltaAla, (Z)-DeltaAbu, (Z)-DeltaLeu, (Z)-DeltaPhe and DeltaVal, to determine a predominant solution conformation of these alpha,beta-dehydropeptide-related molecules. Measurements were taken in CCl4, DCM and MeCN solutions. In the same way, spectra of saturated analogues Ac-Xaa-NMe2, where Xaa = Ala, Abu, Leu, Phe and Val, were investigated. To help interpret the spectroscopic results, conformational maps were calculated by the B3LYP/6-31+G** method. Also, the relative energies of all conformers of the dehydro compounds in vacuo as well as in the studied solvents in addition to the theoretical IR frequencies of these conformers were calculated. For comparison, molecules of two saturated analogues, Ac-L-Ala-NMe2 and Ac-L-Phe-NMe2, were calculated in a similar way. Both unsaturated and saturated compounds, which have an aliphatic side chain, occur in CCl4 and DCM mainly as a mixture of extended conformers with the C5 H-bond and open conformers. As solvent polarity increases, participation of the open conformers also increases, and in MeCN, the model amides are almost exclusively in the open form, except Ac-DeltaAla-NMe2, which shows a small amount of the H bonded conformer. Ac-DeltaAla-NMe2 and Ac-DeltaAbu-NMe2 have stronger C5 hydrogen bonds than those of their saturated counterparts. As the calculations indicate, the open conformation of the unsaturated amides is conformer H/F with phi, psi 44 +/- 5 degrees, 127 +/- 4 degrees. This is the second lowest in energy conformer in vacuo and in CCl4 and the lowest one in more polar solvents. The open conformation of Ac-L-Ala-NMe2 constitutes conformer C with phi, psi -101.5 degrees, 112.7 degrees. For Ac-DeltaAla-NMe2 and Ac-DeltaAbu-NMe2, FTIR also reveals the presence of a third conformer. Calculations indicate that is the semiextended conformer D with the N1-H1...N2 hydrogen bond/contact. In all solvents, Ac-L-Phe-NMe2 and Ac-(Z)-DeltaPhe-NMe2 show only the extended E and the open H/F, respectively. In both there is an amide/pi(Ph) interaction. PMID- 15782430 TI - Theoretical study of 31P, 31P coupling constants in cyclotriphosphazenes. AB - Several scalar coupling constants (mainly 31P, 31P) were calculated for 10 cyclotriphosphazenes and compared with experimental results when available. Although the experimental values cannot be reproduced, the calculated values are proportional to the experimental values. Some difficult cases, such as 19F, 19F couplings, are discussed. PMID- 15782431 TI - High alcohol intake linked with heart-rhythm disorder. PMID- 15782432 TI - Genetic testing kit on the horizon for heart disease. PMID- 15782433 TI - Ask the doctors. Are drug-coated stents living up to potential that you and other publications wrote about a year or so ago? PMID- 15782434 TI - Ask the doctors. A relative of mine may need angioplasty. Is it getting to the point where even small hospitals can do this well? She'd like to be close to home. PMID- 15782435 TI - Bibliography. Current world literature. Nutrition and the intensive care unit. PMID- 15782436 TI - Tongue lesions. Diagnosis: Primary herpetic gingivostomatitis. PMID- 15782437 TI - Diagnosing necrotising fasciitis. PMID- 15782438 TI - [Late onset of bipolar affective disorder?]. PMID- 15782439 TI - [Affective disorder in tertiary syphilis]. PMID- 15782440 TI - Quantitative structure activity relationship studies of diaryl furanones as selective COX-2 inhibitors. AB - Selective COX-2 inhibitors have attracted much attention in recent times in the design of non-steroidal anti-inflammatory agents (NSAID), which are devoid of the common side effects of classical NSAIDs. QSAR studies have been performed on a series of diaryl furanones that acts as selective COX-2 inhibitor using Molecular Operating Environment (MOE). The studies were carried out on 43 analogs. These studies produced good predictive models and give statistically significant correlations of selective COX-2 inhibitory with physical property, connectivity and conformation of molecule. Also when available COX-1 inhibitory data was analyzed with descriptors obtained from MOE, partial charge descriptor, van der Waal's surface area and solvation energy gave statistically significant results. PMID- 15782441 TI - A report from the Society for Women's Health Research. PMID- 15782442 TI - Social and ethical challenges of prenatal diagnosis. PMID- 15782443 TI - Idiopathic arm pain. PMID- 15782444 TI - [M Baltazard, field epidemiologist (1908-1971)]. PMID- 15782445 TI - Complementary and alternative medicine: personal preference or low cost option? AB - From acupuncture to yoga, Americans' use of complementary and alternative medicine (CAM) is widespread and growing. The reasons that people give for using CAM are as diverse as the CAM therapies themselves: some perceive that conventional health care is ineffective, while others consider CAM to be more consistent with their own values and beliefs about health. As conventional health care costs rise, it is also possible that some people turn to CAM as a low cost alternative. This Issue Brief summarizes research that evaluates the relationship between CAM use and perceived access to conventional health care. PMID- 15782446 TI - The influence of lifestyle on cardiovascular risk factors. Analysis using a neural network. AB - The cardiovascular pathologies are the most common causes of death in the elderly patient. To single out the main risk factors in order to effectively prevent the onset of the disease, the authors experimented a special computerized tool, the neural network, that works out a mathematical relation that can obtain certain data (defined as output) as a function of other data (defined as input). Data were processed from a sample of 276 subjects of both sexes aged 26-69 years old. The output data were: high/low cholesterolemia, HDL cholesterol, triglyceridemia with respect to an established cut-off; the input data were: sex, age, build, weight, married/single, number of children, number of cigarettes smoked/day, amount of wine and number of cups of coffee. We conclude that: (i) a relationship exists, deduced from a neural network, between a set of input variables and a dichotomous output variable; (ii) this relationship can be expressed as a mathematical function; (iii) a neural network, having learned the data on a sufficiently large population, can provide valid predictive data for a single individual with a high probability (up to 93.33%) that the response it gives is correct. In this study, such a result is found for two of the three cardiovascular risk indicators considered (cholesterol and triglycerides); (iv) the repetition of the neural network analysis of the cases in question after a "pruning" operation provided a somewhat less good performance; (v) a statistical analysis conducted on those same cases has confirmed the existence of a strong relationship between the input and the output variables. Therefore the neural network is a valid instrument for providing predictive in a single subject on cardiovascular pathology risks. PMID- 15782447 TI - Injury and poisoning episodes and conditions: National Health Interview Survey, 1997. AB - OBJECTIVE: This report provides a descriptive overview of the first year of data from the injury section of the redesigned National Health Interview Survey. It documents the Survey's design methodologies and presents detailed national estimates of nonfatal injury and poisoning episodes for 1997. METHODS: Data for the U.S. civilian noninstitutionalized population were collected using Computer Assisted Personal Interview (CAPI). The data on all medically attended injuries and poisonings occurring to any family member during the 3-month period prior to the interview were obtained from an adult member of the family. RESULTS: In 1997, 34.4 million medically-attended episodes of injury and poisoning were reported, resulting in an age-adjusted rate of 128.9 episodes per 1000 persons. Injury episodes resulted in 40.9 million injury conditions for a rate of 153.7 conditions per 1,000 persons. Falls were the leading external cause of injury, followed by episodes resulting from being struck by or against a person or an object, transportation, overexertion, cutting and piercing instruments, and poisoning. Sprains and strains were the most frequently reported injury condition followed by open wounds, fractures, and contusions. Upper extremity and lower extremity injuries were the leading body regions for these conditions. Leisure activities and paid work were most often reported as the activities the person was engaged in when the injury episode occurred, and. the home was the most likely place for the injury to have occurred. CONCLUSION: The redesigned NHIS is a useful source of information about medically-attended nonfatal injuries and poisonings. A single year now provides enough data to produce stable national estimates on details of injury and poisoning episodes. PMID- 15782448 TI - Current estimates from the National Health Interview Survey, 1996. AB - OBJECTIVES: This annual report presents national estimates, based on data from the National Health Interview Survey (NHIS), on the incidence of acute conditions, percent of medically attended acute conditions, number of disability days, episodes of persons injured and associated activity restriction, persons with activity limitation due to chronic conditions, restricted activity days associated with acute and chronic conditions, physician contacts and short-stay hospitalizations, as well as prevalence of chronic conditions and respondent assessed health status. This edition includes a section on trends in health statistics for 1982-96. SOURCE OF DATA: NHIS is a multistage probability sample survey conducted annually by interviewers of the Bureau of the Census for the National Center for Health Statistics. Data is collected during in-home interviews of the civilian noninstitutionalized U.S. population. Data collection procedures were similar from 1982 through 1996, but were changed after 1996. HIGHLIGHTS: In 1996 there were 163.5 acute conditions per 100 persons, (67.9% were medically attended) and 624.0 associated days of restricted activity per 100 persons. Of acute injuries, 91.4% were medically attended. The highest rates for chronic conditions per 1000 persons included arthritis (127.3), sinusitis (125.5), deformity and orthopedic impairment (111.6), and high blood pressure (107.1). Activity limitation due to chronic conditions was reported by 14.4% of persons. There were six physician contacts per person per year and 7.3% of the population had a hospitalization in the past year. During 1982-96, the prevalence of asthma increased and the rate and duration of hospitalizations decreased. PMID- 15782449 TI - The pathogenesis of defective hemostasis in abruptio placentae. PMID- 15782450 TI - Changes in the fibrinolytic enzyme system following intravascular coagulation induced by thrombin and endotoxin. PMID- 15782451 TI - Metyrapone response in pregnancy. PMID- 15782452 TI - A simple rapid assay of estrogens in pregnancy. PMID- 15782453 TI - Convulsive toxemia. A report of 168 cases managed conservatively. PMID- 15782454 TI - Effect of posture upon sodium excretion in pre-eclampsia. PMID- 15782455 TI - Drug metabolizing systems in homogenates of human immature placentas. PMID- 15782456 TI - Effect of epinephrine on placental carbohydrate metabolism. PMID- 15782457 TI - Radiation to the fetus in intrauterine transfusion. PMID- 15782458 TI - Hereditary factors in the Stein-Leventhal syndrome. PMID- 15782459 TI - Ultrastructure of a granulosa cell tumor. PMID- 15782460 TI - Enzymic histochemical reactions in two Krukenberg tumors associated with clinically different endocrine patterns. PMID- 15782461 TI - Mesonephric carcinoma of the ovary producing hypercalcemia. PMID- 15782462 TI - Genital infection with TRIC agents in Taiwan. PMID- 15782463 TI - Ruptured tuboovarian abscess. PMID- 15782464 TI - Defective development and reproductive wastage in the United States. PMID- 15782465 TI - Biologic effects of intrauterine devices. PMID- 15782466 TI - Summary health statistics for U.S. children: National Health Interview Survey, 1997. AB - OBJECTIVE: This report presents statistics from the 1997 National Health Interview Survey on selected health measures for sample children under 18 years of age, classified by sex, age, race/ethnicity, family structure, parent's education, family income, poverty status, health insurance coverage, place of residence, region, and current health status. The topics covered are asthma, allergies, learning disability, Attention Deficit Disorder, use of medication, respondent- assessed health status, school-loss days, usual source of medical care, time since last contact with a health care professional, selected health care risk factors, and time since last dental contact. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U. S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized population of the United States. Data are collected during face-to-face interviews with adults present at the time of interview. Information about children is collected for one sample child per family in face-to-face interviews with an adult proxy respondent familiar with the child's health. HIGHLIGHTS: In 1997, most U.S. children under 18 years of age enjoyed excellent or very good health (82%). However, 13% of children had no health insurance coverage, and 6% of children had no usual place of medical care. The percent of children with unmet medical need was twice as high in single-mother families (4%) as in two-parent families (2%). Eleven percent of children had ever been diagnosed with asthma. An estimated 8% of children 3-17 years of age had a learning disability, and an estimated 6% of children had Attention Deficit Disorder. PMID- 15782467 TI - [Ethical problems faced by in managing HIV in workplace]. AB - This study which was conducted between September 1999 and June 2000 among occupational physicians aimed to appreciate ethical problems faced by physicians in managing HIV infection in workplace. A questionnaire was administrated by telephone to physicians included in the list of Occupational Physicians Association. Thirty eight physicians responded among the fourty four contacted. Thirty two ie 84.21% did face HIV infection in their workplace. The frequency of meeting cases was in between 1 to 5 for 68.75%. Thirty two did order the test either to depist or to conduct eatiologic diagnosis. Informed consent were asked by twenty one physicians. Six physicians ie 18.75% did inform the employer of serologic status, among them four said having obtain employee's consent to reveal status. For them, such revalation was the basis for financial involvment of employer in the treatment, and facilitates his tolerance of absenteeism. Four physicians signaled facing dilemma when a patient did not want to inform his or her partner of his (her) serologic status. They suggested a legal authorization to reveal patients status in such circumtances. PMID- 15782468 TI - [Seroprevalence of toxoplasmosis among women having spontaneous abortion and pregnant women following in a center of health up-town in Dakar]. AB - A prospective survey carried on 70 women having a spontaneous abortion and 70 pregnant women was achieved in the center of health Roi Baudouin of Guediawaye (Senegal) between November 2001 to April 2002 to study the toxoplasmosis in the two groups. Anti-toxoplasmosis antibodies were searched for on all women by a technical solid-phase enzyme immunoassay. The solid-phase is a combs. The seroprevalence of the toxoplasmosis is 37.1% among women having aborted and 40% for pregnant women. The difference is not statistically significant (p = 0.729). The seroconversion is 22.8% among women having aborted and 10% for pregnant women. The difference is statistically significant (p = 0.041). The research of factors favoring the infestation, related to through food habits, pleads in favor of a contamination during childhood, essentially soil and/or eating undone meat. With regard the high percentage of not immunized women (62.9% for women having aborted and 60% for pregnant women), it is necessary to take prophylactic measures to prevent congenital toxoplasmosis. PMID- 15782469 TI - [Primary lymphoma of the thyroid: a case report]. AB - Primary lymphoma of the thyroid is an uncommon disease. It diagnosis is difficult without immunohistochemestry. We report a case of thyroid lymphoma diagnosed at the A. Le Dantec Hospital of Dakar. A 22-year old young man presented anterior neck mass. Clinical examination showed a thyroid mass with a susclavicular palpable lymph node. The scintigraphy demonstrated that the mass was cold. Total thyroidectomy with cervical lymphadenectomy was the first treatment. Histological examination of the surgical specimen concluded that the mass was an anaplastic lymphoma (large cells type) involving the sus clavicular lymph nodes (stage IIE). Chemotherapy was administrated after one month with C-MOPP schedule. The patient presented no recurrent sign after 30 month. Fine needle aspiration and immunocytochemestry of the thyroid cold masses will contribute in an accurate diagnosis and the management of disease in our institution. PMID- 15782470 TI - [Primary focal segmental glomerulosclerosis in adults: response to treatment and outcome]. AB - Response to therapy of primary focal segmental glomerulosclerosis (FSGS) in adults is poor, most studies demonstrated no response at all. We report our experience from a retrospective study about 22 adults patients. All patients presented proteinuria at admission with a nephrotic syndrome in 86 percent of cases. A high blood pressure was noted in 25 percent. A response to corticosteroid was obtained in 8 patients (36%) with complete remission in 5 cases (23%). Three patients were corticosteroid-dependant, two out of them went in remission under cyclophosphamide in one case and under chlorambucil in the other case. Fourteen patients did not respond to corticosteroids. Eight received cyclophosphamide with partial remission in 4 cases and progression to chronic renal failure (CRF) in 4 cases. The six non-responders did not receive immunosuppressive drugs because of severe CRF in 4 cases. The mean duration of follow-up was 39 months (20-55). The response to corticosteroid of primary FSGS in adults remain low (30%). The use of cyclophosphamide has improved the global remission to 50 percent. PMID- 15782471 TI - [Treatment of incisional hernias]. AB - Several therapeutic processes were proposed in the repair of incisional hernia, on the basis of simple joining suture go to the installation of prosthetic mesh while passing by aponeurotic autoplasty according to Welti-Eudel technique. The aim of this study was to report the results of our experiment in the treatment of incisional hernias. It was a retrospective study carried out of January 11th 1996 to December 31, 2000, concerning 35 cases of incisional hernias operated during the study period. The following parameters were studied: age, sex, diameter, technique of repair, morbidty, mortality and the remote follow-up. Average age of our patients was 33 years with extremes of 13 and 53 years. It include 30 womens and 5 mens. The initial operation were dominated by Caesarean (57%). The diameter of incisional hernia varied between 3 and 5 cm in 22.9% of patients, between 5 and 10 cm in 62.8% of paitents, higher than 10 cm in 14.3% of patients. The simple joining suture was carried out in 22.9% of cases, the Welti-Eudel technique in 42.9% of cases and the installation of prosthetic mesh in 34.2% of the cases. Mortality was 5.1% and the morbidity was 34.2% made exclusively by suppuration including 5.1% on prosthetic material. We noted 14.3% of recurence which has occured after repair by simple joining. The Welti-Eudel procedure gives good results in the repair of incisional hernia with small and average dimensions. The installation of prosthetic mesh constitute the treatment of choice because the rate of recurence is weak even null. PMID- 15782472 TI - [Apical foramen position on mesio buccal root of the maxillary first molar. Study in Senegalese population]. AB - The maxillary first molar is submitted to frequent endodontic treatment and more over, it presents the highest failure rates, often in relation with his mesio buccal root. Several studies about failures factors showed implication of root canal anatomy often in relation with the frequency of second canal. However, other failures factors, as the foramen situation, which is very important in endodontic practice, could increase the failure rate. The aim of this study was to determine the foramen situation in Senegalese people. The results showed 63.85% of excentric foramina (83 out of 130). 86% of excentric foramina were located at 1.5mm radiology apex. PMID- 15782473 TI - [Secondary hyperparathyroidism of chronic hemodialysis]. AB - Secondary hyperparathyroidism is defined as autonomic secretion of parathormon (PTH) whose 1-84 fraction level is up to 60 ng/l. The aim of this study was to determine incidence of secondary hyperparathyroidism on patients undergoing hemodialysis in Le Dantec Hospital of Dakar, and describe its diagnostic and therapeutical aspects. Between 22 patients followed in chronically hemodialysis, 11 patients had effective blood test of parathormon. Seven of them had high level of PTH and the other had normal rate. They were 5 men and 7 women with a mean age of 56 years. The mean duration undergoing hemodialysis was 42 months. Clinical signs were rare and non specific, dominated bony pains and anemia. Ectopic calcifications had been found in 3 cases. Hypocalcemia was present in 5 cases and high level of phosphoremia in all cases. The D3 vitamin was at a normal rate in all cases up to 10 ng/ml. Calcium supplementation with 1.5 to 2.5 g/day was effective associated with rich calcium dietary. This level of calcium supplementation appeared too low even though it must be closely estimated because of the possibility of improving ectopic calcifications. At the opposite, D3 vitamin supplementation seems to be unuseful under tropical areas. PMID- 15782474 TI - [Spiral CT in aortic dissection]. AB - Patients with suspected dissection of the thoracic aorta require immediate diagnostic evaluation so that urgent therapeutic interventions can begin. Two women with suspected aortic dissection and one man with abdominal aortic aneurysm underwent color-flow Doppler echocardiography and CT angiography with multiplanar reconstructions. Spiral CT showed ascending aortic dissection (type A Stanford) in two cases and descending aortic dissection (type B Stanford) in one case. It also access the visualization of the intimal flap, the extent of dissection, the size of the aorta, and the presence of pericardial fluid. CT angiography is an accurate diagnostic procedure for patients with suspected aortic dissection and possesses several advantages over other modalities such as transthoracic echocardiography, transesophageal echocardiography and aortography. PMID- 15782475 TI - [Solitary ulcer of the rectum. Report of two cases treated by the Orr-Loygue technique]. AB - We report two cases of solitary ulcer of the rectum treated by Orr Loygue rectopexy technique in a ten-year period at the Surgery Department of Aristide Le Dantec. The first case is about a 39 year-old female who was constipated since approximately seventeen years and had a rectal prolapse associated later to bloody stools. In 1990, she underwent haemorrhoids surgery operation with unsatisfactory results. After the recurrence of the disease, she was admitted at Aristide le Dantec Teaching Hospital, rectoscopy revealed a solitary ulcer. Orr Loygue rectoplexy technique was performed by laparotomy. In the follow up, there was no complication. The second case is about a 43 years old female who suffered from rectal syndrome since twenty years and had pelvis pains with bleeding and phlegmatic diarrhoeas. The physical examination and biologic analysis showed no particularity. Rectoscopy has revealed a solitary ulcer of the rectum which was confirmed by biopsy. Orr Loygue rectoplexy technique was performed. In the follow up, there was a recurrence and this was confirmed by rectoscopy. The rectopexy alone does not summurize the therapeutic. A final dyskesia or an associated perineal insufficiency must be integrated in a total therapeutic strategy. PMID- 15782476 TI - [The para-umbilical block in children: 75 cases report]. AB - The objective of this study is to assess both intra and post operative analgesia in infants undergoing umbilical hernia repair under general anaesthesia with neither opioid nor muscle relaxant, associated with a para umbilical block. It's a prospective study covering a 15 months period. The study included 75 infants (age = 5 months - 13 years; body weith = 6 kg - 35 kg). General anaesthesia was induced with either thiopentone or halothane and, maintained with halothane in a N2O - O2 50 VOL % mixture. Para-umbilical block was obtained using 1 ml/kg of 0.25% marcaine. Pain was assessed using time course of respiratory rate, heart rate and mean arterial pressure. A change of more than 20% in one of these variables was considered criterion of poor analgesia. Intraoperative analgesia was adequate in all patients but four, 5 minutes after incision. Surgical conditions were considered as being godd or satisfactory in 90.6% and 9.4% of cases, respectively. Post operative analgesia, assessed 1 and 6 hours after completion of surgery was convenient in 93.3% of infants. The block appears as simple, most after efficient and safe in umbilical surgery. PMID- 15782478 TI - [Quality of butter sold in Dakar (Senegal)]. AB - Among foods that are introduced in Senegal, butter takes a significant place particularly at home where it is frequently eaten. Nevertheless, Senegalese authorities don't carry out any quality control under those different types (mark) of butter. In consequence, to find butter without quality or characteristics of authentic butter may be high. The aim of this work is to analyse quality of main varieties of butter in Dakar market. Chemical methods were used to determine the characteristic of butter samples and gas chromatography was made to authentify their fatty acid profile. Our results have shown that most of analysed sample respected the standards of quality. In fact the composition of fatty acid and characteristic factors of authentic butter were certified by this study. PMID- 15782477 TI - [Sodium fluoride controlled release system for intra buccal use]. AB - Hydroxyapatite which has the same elemental chemical composition as natural bone and teeth is one of the promising raw material for the design of drug controlled release system in intrabuccal use. It is stable and biocompatible and widely used in orthopedics and odontology. So, in order to improve the administration of drugs for intrabuccal use, we have developped a fluoride controlled release delivering system. We have formulated tablets of 160 to 200 mg to be fixed on the vestibular face of a molar or a premolar and permitting to reach high enough local concentrations for desirable therapeutic effect. The tablets have a granular matrix composed of hydroxyapatite and fillers, ethylcellulose and/or Eudragit. For all tablets, the pharmacotechnical values support the pharmacopoeia norms. On fragments of tissue maintained in culture, the sodium fluoride is released at constant rate. The release profiles observed are predictable. No disintegration of tablets have been observed during release studies and after. Histological analyses performed after release studies showed the biocompatibility qualities of the tablets. PMID- 15782479 TI - [The ovarian tumors in children. A report of 12 cases]. AB - The objectives emphasize the diagnosis and therapical problems concerning the child's ovary tumors for a better care in our conditions of work. We collected in a retrospective study 12 cases of ovarian tumors during 30 years. The middle age of the patients was of 11 years with extremes of 2 years and 15. The circumstances of discovery of the tumor were represented by order of frequency by an abdominal tumor (10 cases), an abdominal pain (8 cases) and of the compression signs digestive or urinary (6 cases). The medical imagery (echography, intravenous urography, Chest X-ray) permitted to evoke in the majority of the cases the ovarian origin of the mass. The surgical treatment consisted in an ovariectomy (5 times), an salpingo - ovariectomy (6 times) and a surgery of partial exeresis associated to a chemotherapy. The operative continuations were simple. A patient died of intercurrentes complications. The histological exam of the operative piece showed that it was about benign tumors. However a brought closer surveillance proves to be necessary had consideration to the two noted recidivisms. PMID- 15782480 TI - [HIV associated nephropathy syndrome: a case report in Dakar]. AB - HIV associated nephropathy syndrome ( HIVAN Syndrome ) is a recently identified entity and no study has been done in Senegal. So we report this observation. A 40 years old black patient was admitted for renal oedema syndrome and immunosuppressive signs. The biological investigations noticed a non-pur nephrotic syndrome and severe renal failure. Ultrasonography showed quite normal kidney sizes with hyper echogenicity and dediferenciation. HIV research was positive with 45 CD4 lymphocytes / mm3. This patient had no known causes of nephrotic syndrome ( diabetis, lupus ,amyloidosis.). So the diagnosis of HIVAN syndrome was determined with the clinical features and the bad outcome. We emphasize on the necessity to think about HIVAN in every black patient presenting a quickly progressive non-pur nephrotic syndrome. We expect prospectives studies to describe the clinical signs and the frequency of HIVAN syndrome in Senegal. PMID- 15782481 TI - [Evaluation of oral health and access to care in senegalese children with Down syndrome: preliminary study]. AB - Down syndrom is the most frequent genetic disease discribed for viable (live born) children and the commonest cause of mental deficiency in developed countries. It is an autosomal chromosomic anomaly resulting from trisomy of all or part of chromosome 21 and is characterizd by general and mental deficiencies and a typical facial phenotype. The aims of this study are:--to evaluate the general and oral health of a senegalese group of children with Down syndrome and their siblings the closest in age,--to assess their access to care. We used the OADS (Oral Assessment Down Syndrome) questionnaire in 25 senegalese children with Down syndrome living in Dakar and 17 od their siblings the closest in age. Their ages range from 4 to 20 years old in both groups. Our results revealed that senegalese children with Down syndrome have more general and oral health problems compared to their siblings the closest in age but they were more likely to have difficulty finding both medical and dental services. This study pointed out the need for special care services with trained people for a better management of these specific patients. PMID- 15782482 TI - [Nasal carriage of meticillin-resistant Staphylococcus aureus among health care personnel in Abidjan (Cote d'lvoire)]. AB - To determine the prevalence of meticillino-resistant Staphylococcus aureus (MRSA) among health care personnel in Abidjan teaching hospitals as well as their resistance profile against other antibiotics, 592 health care personnel from various surgical and medical services: the intensive care unit, gynaecology and obstetrics and third-degree burns services of the Cocody, Treichville and Yopougon Teaching Hospitals were included. The previous nasal pits of each subject included were swabbed. The isolation of S. aureus strains was run in a Chapman medium followed by Identification based on morphological and biochemical characteristics. The resistance profile of the strains to antibiotics was determined by standard Kirby-Bauer disk diffusion method and a 1 microg disc of oxacillin was used for the detection of meticillin-resistance S. aureus strains according to NCCLS (National Committee for Clinical Laboratory Standards) guidelines. 269 members of the studied personnel were carriers of S. aureus, either a rate of portage of 45.4%. Among the 269 S. aureus isolates, 38.7% were MRSA strains and the carriage rate of MRSA in the population was 17.8%. The health care personnel working in surgery was the more colonized (36.7%) follow-up of those of the medical services (31.4%) and of the the intensive care unit (12.4%). A variable proportion of strains of MRSA also expressed resistances to the other families of antibiotics: 27% to aminosids of which 13.5% of phenotype kanamycine, tobramycine, gentamycine (KTG), 58.7% to macrolids and related (MLS), 37.5% to fluoroquinolons, 14.4% to cyclines and 40% to the cotrimoxazole. This confirms their multi-resistant character. The prevalence of MRSA carriage among health care personnel is high; this personnel constitutes an infectious risk for the hospitalized patients who are so exposed to nosocomial infections caused by MRSA. PMID- 15782483 TI - [Diffculties in medico-social management of tuberculosis in Pneumology Departement of Fann University Teaching Hospital]. AB - This prospective study included all the patients who, during the month of September 1995, were admitted for pulmonary baclliferous tuberculosis to the Pneumology Clinic of the Fann University Hospital, Dakar. The patient's escorts and the health personnel were also included in this study. The aim of the study was to find the different socio-economic and health factors impeding the hospitalization of tuberculosis patients in the Pneumology Clinic of Fann University Hospital. 22 members of the health team, 209 patients aged between 15 and 65 years and 209 escorts were interviewed. Out of the 10% of tuberculosis patients with HIV seropositivity, 80% admitted not to have informed their spouses of their infection. The decision to be admitted was made by the patient himself in 54.4% of cases and by his family in 45.5% of cases. Late admittance with regards to the beginning of symptoms was due to the recourse to traditional medicine in 43.7% of cases, wrong diagnosis in 24%, ignorance in 19.3% and, in 13% of cases, due to inappropiate anti tuberculosis treatment. Despite the fact that anti tuberculosis medicine was free of charge, each patient or his family spent an average sum of 87,500 CFA F (US dollar 175) for a month's admission (the minimal salary (SMIG) in Senegal is 32,000 CFA F (US dollar 64), and 40% of the patients and escorts had difficulties making this payment. 9 patients were judged to be poor by 25.7% of the patients and 8.7% found relations with the health personnel difficult. The escorts deplored the lack of toilets (only 1 out of 4 was functional), the time worn facilities, the overcrowding as well as the irregularity and poor quality of the hospital meals. The entire health team deplored the lack of adequate personnel and 30% of them deplored the lack of hygiene of some patients and escorts. Taking financial charge of tuberculosis patients at the Pneumology Clinic of the Fann University Hospital requires an increased financial effort from the State (rehabilitation of the facilities, recruitment of medical and paramedical personnel, improved meals) harmonisation on a national scale of anti tuberculosis therapeutic protocoles and an Information-Education-Communication (IEC) programme on tuberculosis and hygiene. PMID- 15782484 TI - [Surgical management of brain-stem gliomas: a retrospective analysis of 311 cases]. AB - OBJECTIVE: To further study the clinical features, diagnosis, and surgery outcome of brain-stem gliomas. METHODS: Totally 311 patients with brain-stem gliomas received surgery operations in our hospital from 1980 to the end of 2001. The clinical data, neuroimages, growth patterns, and operative feasibility were analyzed. RESULTS: Different brain-stem gliomas showed different growth patterns. In this series, total excision rate of the tumor was 40.5%, subtotal 29.9%, partial 29.6%, and operative mortality 1.3%. Among 311 patients, 72.4% patients had been improved and stable in their symptoms after operation, and 27.6% deteriorated or having transitory new signs. Five years survival rate is 67% in ependymoma patients, 42% in astrocytoma patients. None of brain-stem glioblastoma patients survived up to 5 years. CONCLUSIONS: The histology and growth pattern of brainstem gliomas varies. The patients with well-differentiated gliomas of brain stem may be cured by microsurgical removal. For malignant ones, partial removal may prolong survival and facilitate the following combined therapy. PMID- 15782486 TI - [Gamma knife radiosurgery for cavernous malformations]. AB - OBJECTIVE: To evaluate the efficacy of Gamma knife surgery (GKS) in treating cavernous malformation (CM). METHODS: From 1994 to 2001, 92 patients with 114 CMs were treated by GKS and then followed up for 2-8 years (mean 4.1+/-1.9). We analyzed the MRI features of CMs bleeding, efficacy of GKS, and the complications of treatment. Six pathological specimens after radiosurgery (1 from our group, 5 from other centers) were also assayed. RESULTS: Among 43 patients who were treated by GKS to control their epilepsy, epileptic paroxysm was alleviated in 36 patients (83.7%), including 12 (27.9%) seizure-free. Rebleeding was confirmed in 9 patients (9.8%) by neuroimage, one of whom died. Transient symptomatic radiation edema occurred in 7 cases (7.6%) within 6-12 months after radiosurgery, and one patient underwent open surgery for cerebral decompression. The main pathological changes of cavernoma were coagulation necrosis and the vessels obliterated gradually after radiosurgery. CONCLUSIONS: It is feasible to treat small and surgically high risk CMs by radiosurgery. The treatment has to be prudent in an acute bleeding and symptomatic progression. Optimal treatment timing and dose planning are prerequisites to reduce radiation-related complications. GKS is safe and effective to control the epilepsy caused by CMs, and also to bring down the rebleeding rate after a latency interval of several years. PMID- 15782485 TI - [Relationship between invasive pituitary adenomas and cavernus sinus and cariod artery and surgical outcomes]. AB - OBJECTIVE: To analyze the relationship between invasive pituitary adenomas and cavernus sinus and cariod artery and to predict their surgical outcomes. METHODS: Totally 270 patients with pituitary tumors were investigated in this retrospective study, including 113 men and 157 women, with a mean age of 40.8 years. The mean disease history was 3.6 years. Pituitary microadenomas were diagnosed in 56 cases, macroadenomas in 118 cases, and hugeadenomas in 96 cases. Adrenocorticotropic hormone-releasing adenomas (Cushing's diseases) were diagnosed in 40 cases, growth hormone-releasing adenomas in 58 cases, prolactinomas in 57 cases, and non-functional pituitary adenomas in 115 cases. Transsphenoidal microsurgery were performed on 260 patients, while transcranial microsurgery on 6 patients. RESULTS: The percentage of invasive pituitary adenomas was about 3.6% in microadenomas, 20.4% in macroadenomas, and 61.4% in hugeadenomas. Rate of total removal was 94.1% in non-invasive pituitary adenomas, and was 58.8% in invasive pituitary adenomas. CONCLUSIONS: It is important to analyze the grade of invasive pituitary adenomas to improve the removal of pituitary adenomas, avoid injuring cariod artery, and increase the rate of total removal. PMID- 15782487 TI - [Clinical application of minimally invasive neuroendoscopic techniques]. AB - OBJECTIVE: To summarize and analyze the application of neuroendoscopic techniques in neurosurgery, and to discuss the role and significance of neuroendoscopic techniques in the diagnosis and treatment of neurosurgical diseases. METHODS: We treated 1300 patients with different neurosurgical diseases by performing endoscopic neurosurgery (EN) and endoscopy-assisted microneurosurgery (EAM). Among 1300 paitents, 522 were treated with pure endoscopic neurosurgery, 260 with endoscopy-assisted microneurosurgery, 79 with endoscope-controlled bur hole trephination neurosurgery, 434 with endoscope transsphenoidial surgery, and 5 with other techniques through which an endoscope was used in conjunction with stereotactic guidance. RESULTS: Totally 362 hydrocephalus patients were treated using EN. Among them, 190 were treated by third ventriculostomy, 30 by V-P shunt, and 142 patients with complicated hydrocephalus and unsymmetry hydrocephalus by endoscopy-controlled pathologic septum fenestration, septum pellucidum fenestration, and treatment of inventricula inflammation. Clinical symptomatic improvement was achieved in 341 of 362 patients (94.2%). Also 160 intracranial cyst patients were treated using EN for resection and partial resection. Eighty two patients were performed through cyst-ventricula fenestration. Clinical symptomatic improvement was achieved in 76 of 82 patients (92.7%). Seventy patients treated with endoscopy-controlled bur hole neurosurgery and 8 cases with endoscopy-assisted microneurosurgery got better recovery after operation. Among 260 patients with brain tumors, 252 patients were operated with EAM (190 patients with epidermoid cyst), 8 patients with EN (all brain tumors with diameters < 2.5 cm in inventricular). Clinical symptomatic improvement was achieved in 228 of 260 patients (87.7%). Among 49 patients with inventricular and cistern cyst, 40 patients who were treated by EN and 9 patients by endoscopy-controlled bur hole neurosurgery were resected and their clinical symptoms were improved after operation. Among 434 patients with sellar region lesions, 387 patients with pituitary adenomas, 19 patients with repair for CSF leaks, 9 patients with chordoma, and 19 patients with other neurosurgical diseases were performed with endoscopy-controlled transsphenoidial surgery. Clinical symptoms in 88.9% (386/434) of these patients were improved. Another 5 patients were treated with endoscopy combined with navigation and stereotatic guidance with good results. The complications related to operation were found in only 2% of all the patients including hemorrhage, infection, and damage of important structure. CONCLUSIONS: Clinical application of neuroendoscopic techniques can decrease the damage caused by pure open surgery operation. It is possible to resect lesions at the utmost and protect normal tissue from lesions for using EN and EAM or endoscopy controlled microneurosurgery (ECM). It is also helpful to enhance surgical quality and, reduce the complications. PMID- 15782488 TI - [Cerebral revascularization in treatment of intractable aneurysms]. AB - OBJECTIVE: To evaluate the effectiveness of cerebral revascularization in the treatment of intractable aneurysms and to discuss the indications, surgical techniques, and the outcome of the revascularization. METHODS: During the recent 4-year period, 9 radical artery grafts were performed in patients with intractable or giant internal carotid artery (ICA) aneurysms. The indications for cerebral revascularization included parent vessel occlusion during the treatment of the intractable aneurysms with poor collateral circulation or the young patients. Modified techniques were taken to use the main trunk of superficial temporal artery (STA) as donor, while M3 branches near the bifurcation of the M2 segment of the middle cerebral artery (MCA) were chosen as the recipient arteries. STA-Radial artery (RA)-MCA bypass was followed by parent vessel occlusion via chronic cervical ICA ligation or balloon occlusion. RESULTS: Postoperative angiography demonstrated the patency of the grafts in eight cases. The procedure of the parent vessels occlusion was uneventful in these patients. Follow-up showed the patients were in excellent conditions after successful treatment of aneurysms. Significantly delayed filling of the graft was revealed in one patient, who could not tolerate balloon occlusion test and occlusion of parent artery failed. CONCLUSIONS: Extracranial-to-intracranial bypass followed by parent vessel occlusion is a safe and effective method to treat intractable ICA aneurysms. Radical artery as graft can offer high-flow revascularization with less complications. Combined surgical and endovascular treatment might be the future direction for the treatment of the difficult aneurysms. PMID- 15782489 TI - [Correlation between APOA gene polymorphism and intracranial aneurysm]. AB - OBJECTIVE: To study the correlation between a pentanucleotide repeats (PNR) polymorphism of APOA and the genesis of intracranial aneurysm. METHODS: Fifty eight patients with intracranial aneurysms diagnosed by angiography and 58 healthy controls were enrolled in our study. The gene polymorphism of APOA PNR were detected by polymerase chain reaction (PCR) and non-denatured polyacrylamide gels electropherosis (PAGE). RESULTS: Eight kinds of gene types and 6 kinds of alleles were found in these two groups. There were two sites of sequence variance in the 5' control region of APOA gene, which were significantly different between the patients and controls. CONCLUSION: Correlation may exist between intracranial aneurysm and APOA gene. PMID- 15782490 TI - [Transvenous occlusion of carotid cavernous fistula]. AB - OBJECTIVE: To evaluate the treatment of carotid cavernous fistula with the transvenous approaches. METHODS: Seventeen patients (14 males and 3 males) with carotid cavernous fistula (CCF) were retrospectively analyzed. RESULTS: Totally 18 cavernous sinus of 17 patients were occluded. Nine operations were performed via inferior petrosal sinus approaches and 6 via the facial vein. One patient (bilateral fistula) received both two approaches. The fistula were completely occluded immediately after the procedures in 12 patients. Small shunts remained in four of other patients (2 pterygoid drainage and 2 inferior petrosal sinus, respectively). Facial vein catheterization failed because of vein vasospasm in one patient; his symptoms relieved 5 days later and angiography showed the CCF disappeared another 2 days later. CONCLUSION: Transvenous embolization of CCF is effective, safe, and easy to perform. PMID- 15782491 TI - [Treatment of severe vertebral compressive fractures in the elderly with percutaneous vertebroplasty]. AB - OBJECTIVE: To evaluate the efficacy of treating severe and chronic vertebral compressive fractures in the elderly with percutaneous vertebroplasty. METHODS: Sixteen patients who suffered from severe back pain and whose daily living was badly affected were retrospectively reviewed. The average age was 72.5 years, the average disease history was 19 months, and the average compressive rate of the affected vertebral bodies was 74.1%. RESULTS: Nineteen affected compressive vertebral bodies in 16 cases were treated with percutaneous vertebroplasty. All the procedures were successful without any complication. After 3.5-7 ml of cement was injected into the lesions, complete relief was achieved in 3 cases, remarkable relief was achieved in 11 cases, and improvement was observed in 2 cases. The scores of 6-point Behavioral Rating Scale and Activity of Daily Living (ADL) declined significantly after the treatment (P < 0.001). CONCLUSION: Percutaneous vertebroplasty can significantly improve the symptoms and quality of life in the elderly patients with severe and chronic vertebral compressive fractures. PMID- 15782492 TI - [Expression of a human myofibrillogenesis regulator 1 gene in E. coli and its immunogenicity]. AB - OBJECTIVE: To study the expression of human myofibrillogenesis regulator 1 (MR-1) gene in E. coli and obtain the MR-1 protein and its antibody for further investigation of its biological function. METHODS: Expression vectors pGEX-5X-1, pET30a (+), and pET24a (+), as well as host strain E. coli BL21 (DE3) and BL21 CodonPlus (DE3) -RIL were used for expression of MR-1. MR-1 N-terminal with GST or T7-tag or C-terminal with His-tag, separately, or N terminal with T7-tag and C terminal with His-tag, simultaneously, were fused in plasmids pGEX-5X-1, pET30a (+) , and pET24a (+). The expressed MR-1-T protein, separated and purified by preparative SDS-PAGE, was applied to immunize the rabbits. The titer of the antibody was assayed by ELISA and its immunogenicity was tested by Western blot with pcDNA3/MR-1 transfected human breast cancer cell MCF7. RESULTS: The MR-1 protein was successfully expressed as inclusion body by fusing its N-terminal with T7-tag in E. coli BL21-CodonPlus (DE3) -RIL. MR-1 protein was purified by electro-elution from SDS-PAGE gel. Using this purified protein, polyclonal antibody in rabbit against MR-1 was essentially generated. ELISA and Western blot showed the titer of this antibody was about 1:10(5) with high immunogenicity. CONCLUSIONS: The N-terminal fusion tag is the most important mechanism for MR-1 expression. The polyclonal antibody of the generated MR-1 protein in E. coli may be applied for its further biological function studies. PMID- 15782493 TI - [Role of procalcitonin in the differentiation and surveillance of systemic inflammatory response syndrome]. AB - OBJECTIVE: To evaluate the discrimination of serum procalcitonin (PCT) and interleukin-6 (IL-6) between patients with sepsis and non-infectious inflammatory response syndrome (SIRS) and the prediction power of clinical outcome. METHODS: A perspective study was performed in 27 patients with sepsis and 30 patients with non-infectious SIRS. The serum concentrations of PCT, IL-6, C-reactive protein (CRP), white blood cell count, percentage of neutrophil, absolute neutrophil count, and maximal body temperature were obtained less than 24 hours after clinical onset of sepsis or SIRS. RESULTS: The serum levels of PCT and IL-6 and percentage of neutrophil were significantly higher in patients with sepsis than in those with non-infectious SIRS (PCT: 5.54 [1.20, 32.74] microg/L vs 0.77 [0.22, 3.90] microg/L, P=0.001; IL-6: 163.66 [33.60, 505.26] ng/L vs 37.72 [22.52, 110.78] ng/L, P=0.004; CRP [15.28 +/- 8.41] g/L vs [9.51 +/- 7.65] g/L, P=0.010; and percentage of neutrophil: 0.91 +/- 0.04 vs 0.88 +/- 0.04, P=0.010). Receiver operating characteristic curve showed that the power of PCT and IL-6 were the best of all above. There was significant correlation between serum PCT or IL-6 and the acute physiology and chronic health evaluation (APACHE II) or sepsis-related organ failure assessment (SOFA) scores, so was between serum PCT and the intensive care unit (ICU) length of stay. CONCLUSIONS: PCT and IL-6 are more reliable indicators to differentiate sepsis and non-infectious SIRS than the conventional inflammatory markers, and correlate with the disease severity. PCT levels are significantly correlated with ICU length of stay. PMID- 15782495 TI - [In vitro vasculogenesis and angiogenesis of mouse embryonic stem cells]. AB - OBJECTIVE: To explore an optional condition to induce mouse embryonic stem (ES) cells to differentiate into endothelial cells and to establish in vitro models of vasculogenesis and angiogenesis. METHODS: Mouse ES cells were cultured in differentiation medium containing a cocktail of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), interleukin-6 (IL-6) and erythropoietin (EPO) in 1% methylcellulose to induce formation of embryoid bodies (EBs). At day 11, EBs were harvested and suspended in rat-tail collagen type I with the same cocktail of cytokines cultured for three additional days. The differentiation of ES cells into endothelial cells, processes of vasculogenesis and angiogenesis were examined using immunostaining of EBs slices and whole-mount immunocytochemistry of EBs with monoclonal antibodies (mAbs) against platelet endothelial cell adhesion molecule-1 (PECAM-1) and alpha-smooth muscle actin (SMA). RESULTS: Under appropriate culture conditions; ES cells spontaneously differentiated and formed EBs containing vascular structures and tubular channels, which were positive for PECAM-1 co-differentiated with smooth muscle. When not treated with angiogenic growth factors, PECAM-1-positive cells could not organize into vascular structures of 11-day-old EBs. In the presence of angiogenic factors 11-day old EBs embedded into type I collagen, and rapidly developed an endothelial networks. Whole-mount immunocytochemistry of collagen gel with anti-PECAM-1 antibody showed the formation of primary vascular structures sprouting from EBs. Quantitative analysis revealed that 100 microg/ml thalidomide significantly reduced the number and length of EBs endothelial sprouting. CONCLUSIONS: Mouse ES cells can differentiate into endothelial cells combined with smooth muscle differentiation during EBs formation and further develop endothelial outgrowths after EBs embedded into collagen, which respectively recapitulate vasculogenesis, angiogenesis, and arteriogenesis processes in vivo. The models provide a useful tool to investigate vasculogenesis, angiogenesis, and arteriogenesis mechanisms and evaluate the effects of angiogenic and angiostatic agents. PMID- 15782494 TI - [Comparison of doxycycline, losartan, and their combination on the expression of matrix metalloproteinase, tissue inhibitor of matrix metalloproteinase, and collagen remodeling in the noninfarcted myocardium after acute myocardial infarction in rats]. AB - OBJECTIVE: To compare the effects of matrix metalloproteinase (MMP) inhibitor doxycycline, losartan, and their combination on the expression of MMP-8, 13, tissue inhibitor of MMP-1, 2 (TIMP-1, 2), and collagen remodeling in the noninfarcted myocardium after acute myocardial infarction (AMI) in rats. METHODS: Two hundred and fifty-four AMI rats, induced by left coronary ligation, were randomly assigned to the following groups: (1) AMI controls group (n = 64); (2) doxycycline group (30 mg x kg(-1) x d(-1), n = 63); (3) losartan group (10 mg x kg(-1) x d(-1), n = 62); (4) concomitant doxycycline and losartan group (30 and 10 mg x kg(-1) x d(-1) respectively, n = 65); and (5) Sham-operated rats (n = 30), which were randomly selected to serve as noninfarction controls. Each group was further divided into three subgroups of 1, 2, and 4 weeks that received treatment. After the completion of treatment, the rats were killed. The mRNA and protein expression of MMPs and TIMPs in the noninfarcted myocardium were quantified by RT-PCR and Western blot, respectively. The type I and type III collagen volume fraction (CVF) of the noninfarced myocardium were assessed immunohistochemically. RESULTS: No significant difference existed in myocardial infarction sizes among the 12 subgroups of AMI controls and the three treatment groups (42%-48%, all P > 0.05). Compared with sham operated rats, the mRNA and protein expression of MMP-8 and 13 significantly increased by 39%-183% in all three subgroups of AMI controls (all P < 0.05), except both of their mRNA expressions in 2-week subgroups; the mRNA and protein levels of TIMP-1 increased only in 1-week subgroup of AMI controls by 104% and 67%, respectively (both P < 0.05); the mRNA of TIMP-2 increased in all 1, 2, and 4-week subgroups by 144% 232% (all P < 0.05), but its protein expression lagged and only enhanced in 2 and 4-week subgroups of AMI controls by 231% and 332%, respectively (both P < 0.05). Meanwhile, both type I and type III CVF of noninfarcted myocardium significantly increased in all three subgroups of AMI controls (type I CVF: 3.01%-5.64% vs 1.53%-1.67%, P < 0.01-0.001; type III CVF: 2.19%-4.42% vs 1.46%-1.59%, P < 0.05 0.001), with type I CVF being higher in 4-week than in 1 and 2-week subgroups (5.64% vs 3.01% and 3.02% respectively, all P < 0.05). Compared with AMI controls, all three kinds of treatment significantly reduced the increased mRNA and protein expressions of MMP-8, 13 and TIMP-1, 2 after AMI by 14%-60% (all P < 0.05), as well as type I/III CVF in their 2 and 4-week subgroups (type I CVF: 1.56%-2.38% vs 3.02%-5.64%, P < 0.05-0.001; type III CVF: 1.92%-2.65% vs 4.19% 4.42%, P < 0.05-0.01), except for doxycycline's effect on type III CVF in any of its three subgroups (all P > 0.05). Among the three treatment groups, significant differences existed in the above mentioned indicators only at some subgroup levels (all P < 0.05). CONCLUSIONS: Like losartan, doxycycline can also suppress the enhanced mRNA and protein expression of MMP-8, 13 and TIMP-1, 2, and reduce type I collagen deposition in the noninfarcted myocardium after AMI in rats. However, it has no effect on type III collagen deposition. PMID- 15782496 TI - [Enhancement of herpes simplex virus-1 glycoprotein-D DNA vaccine induced specific immune responses by coimmunization with interleukin-2 genetic adjuvant]. AB - OBJECTIVE: To investigate the immune responses and protection from virus challenge, induced by the coinjection of IL-2cDNA with herpes simplex virus type 1 (HSV-1) glycoprotein-D (gD) DNA vaccine. METHODS: Two DNA vaccines (pgD and pIL 2) were constructed by inserting the gD gene and IL-2 cDNA into the eukaryotic expression vector pcDNA3.1, respectively. The BALB/c mice were inoculated intramuscularly three times at 2-week intervals. Two weeks after the final immunization, mice were bled for antibody assay and spleen cells were separated for Th cell proliferation and cytokine assays. Delayed type hypersensitivity (DTH) response was detected by the pinna-swelling test. Corneal protection under HSV-1 virus challenge was continuously observed with slit-lamp microscope. RESULTS: IL-2 cDNA coinjection remarkably enhanced the specific IgG2a level when compared with gD plasmid vaccination alone. Th cell proliferation and secretion of cytokines (IL-2 and IFN-gamma) were significantly increased by IL-2 cDNA coinjection. However, the production of IL-10 was inhibited. The DTH response was also enhanced by IL-2 coinjection. When the mice were challenged with HSV-1, the cornea epithelial lesions were significantly alleviated by IL-2 coinjection as compared with gD vaccination alone. CONCLUSION: IL-2 cDNA can enhance both the humoral and cellular immune responses, and thus increase the vaccine potency. PMID- 15782497 TI - [Mechanisms of increased survival rate by administration of AG490 in rats following extensive liver resection]. AB - OBJECTIVE: To study the mechanism by which AG490 improves the survival rate of rats following extensive liver resection. METHODS: Thirty-eight rats were randomly divided into two groups after surgery: control group (n=10), without treatment; (2) AG490 group (n=28), with AG490 (1 mg x kg(-1) x 12 h(-1)) administrated intraperitoneally immediately and 36 hours after the operation. The survival rate was observed and the serum liver functions were measured. RESULTS: The survival rates of control group and AG490 group were 0% and 25%. AG490 group had significantly better blood glucose and aminotransferase levels (P < 0.05) than control group; serum bilirubin levels significantly decreased 48 hours after the operation. Serum protein levels in both two groups had slow decrease but without statistical significance (P > 0.05). CONCLUSIONS: AG490 can significantly increase the survival rate of rats following extensive liver resection. Such a benefit mainly results from the protection towards residual liver function rather than from the promotion of liver regeneration. PMID- 15782498 TI - [Phenotype heterogeneity associated with mitochondrial DNA A3243G mutation]. AB - OBJECTIVE: To discuss the clinical characteristics associated with mitochondrial DNA A3243G mutation. METHODS: Clinical manifestations as well as results of brain CT and/or MRI scanning, blood level of lactic acid and muscle biopsy results of 25 mitochondrial encephalomyopathies patients whose A3243G mutations were analyzed. RESULTS: Although all of the 25 patients carried mtDNA A3243G point mutation, their clinical manifestations varied greatly. Among them, there were 19 cases of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), 2 cases of encephalopathies which could not be classified into any specific type, 2 cases of floppy infants, one case of Kearns-Sayer syndrome (KSS) and one case of mitochondrial entero-myopathy. Most patients showed abnormal cranial radiological findings and ragged-red-fibers on muscle biopsies. Elevation of blood lactic acid was notably found in all of the 25 patients. CONCLUSIONS: Significant variations in clinical manifestation and brain images are the prominent features in patients with A3243G mutation. Mitochondrial diseases should be considered in patients with multiple organ involvement and elevated serum lactic acid mtDNA mutation examination is necessary for the diagnosis of mitochondrial diseases. PMID- 15782499 TI - [Rat alveolar type II injured by bleomycin]. AB - OBJECTIVE: To explore dysfunction mechanism of rat alveolar type II (AT-II) injured by bleomycin (BLM). METHODS: SD rats were injected with a single intratracheal dose of bleomycin or control saline. On day 7, 14, and 28 following intratracheal bleomycin or saline instillation, animals were killed under overdose of 1.5% sodium pentobarbital (0.25 ml/100 g, i.p.) and bronchoalveolar lavage fluid (BALF) from the lung was tested for the activity of pulmonary surfactant (PS) by the Whihelmy Film Balance. Several concentrations of bleomycin stimulated the culture of rat AT-II cells, and surfactant protein (SP) A, B, and aquaporin-1 (AQP) mRNA were analyzed by fluorescent quantitative polymerase chain reaction (FQ-PCR). RESULTS: The activity of PS and hypoxemia significantly decreased on day 7 and improved on day 14 and completely recovered to normal status on day 28. SP-A, B, and AQP-1 mRNA expression in BLM-stimulated group were significantly lower than those in the control group (P<0.001). CONCLUSION: BLM injured AT-II cells decrease the levels of SP-A, B, and AQP-1 mRNA and cause PS dysfunction, resulting in hypoxemia and pneumonedema. PMID- 15782500 TI - [Inhibitory effects of aspirin, clopidogrel, and their combination on the progression of atherosclerosis in rabbits]. AB - OBJECTIVE: To evaluate the effects of aspirin, clopidogrel, and their combination on the progression of aortic atherosclerosis. METHODS: Forty-nine male rabbits were randomly divided into five groups. One group of control rabbits (group N, n=9) was fed on normal diets. Four other groups were fed on high cholesterol diets and injected with 10% albumin bovine received no drug therapy (group M, n=10), aspirin (group A, n=10), clopidogrel (group C, n=10), or the combination (aspirin+clopidogrel, group B, n=10) for 12 weeks. Serum lipids and C-reactive protein (CRP) were detected. The aortas were harvested for histomorphometry and quantitative analysis. The positive percentage of macrophage cells and smooth muscle cells in the plaque were analyzed by immunohistochemistry. RESULTS: These antiplatelet drugs significantly reduced the extent of atherosclerosis. Aortic vascular lesions of group A, C, and B showed 28.70%, 28.82%, and 57.69% of reduction in the amount of macrophage cells, and 39.86%, 42.60%, and 100.70% of increase in smooth muscle cells. No significant differences existed between group A and group C. However, the combination of aspirin and clopidogrel had better effect than aspirin or clopidogrel alone. Meanwhile, aspirin, clopidogrel, and their combination also decreased serum CRP (P < 0.05-0.01), without affecting lipid levels. There was a trend, not significantly, to lower serum C-reactive protein in group B compared with group A or group C. CONCLUSIONS: Aspirin and clopidogrel inhibit neointimal proliferation and prevent the development of atherosclerosis with equivalent effects. Their combination shows synergic effects. PMID- 15782501 TI - [Effects of comprehensive interventions in community on smoking, chronic bronchitis, and asthma in rural areas of Beijing]. AB - OBJECTIVE: To study the effects of comprehensive interventions in community on smoking, chronic bronchitis, and asthma in rural areas of Beijing. METHODS: Twenty-three villages in rural areas of Beijing were randomly divided into interventional (13 villages) and control villages (10 villages) in 1992. Comprehensive interventions including education of former-smokers and improvement of living environment were carried out in the interventional villages, and none was done in the control villages. In April 2000, surveys on smoking, chronic bronchitis, and asthma were carried out among 34,436 participants aged 15 or more in the interventional and control villages. During the same period, knowledge on prevention from chronic obstructive pulmonary diseases (COPD), living environments, and smoking were assessed among 1658 high-risk individuals of COPD at baseline and following-up period. RESULTS: The scores of knowledge and improvement on living environments in the interventional villages were significantly higher than those in control villages (P < 0.001). The decrease rate of smoking and current smoking rate in the interventional villages were significantly higher than in the control villages (0.4% vs -0.8%, P < 0.001; 2.4% vs 1.3%, P < 0.001) in men, while not different in women (P > 0.05). Among never smokers at baseline, the accumulated incidence of smoking among people aged 15 to 24 from 1993 to 2000 was significantly lower in the interventional villages than in the control villages in men (18.9% vs 23.7%, P = 0.005) and in women (0% vs 0.7%, P = 0.005). Daily cigarettes smoked by smokers in the interventional villages were less than in control villages in both men (14.8 +/- 7.0 vs 17.2 +/- 8.2 cigs daily, P < 0.001) and women (12.8 +/- 6.9 vs 13.4 +/- 7.2 cigs daily, P = 0.088). The increase of prevalence of chronic bronchitis in the interventional villages was less than in the control villages (men: 0.9% vs 1.3%, P = 0.012; women: 0.1% vs 0.3%, P = 0.003). After the age factor is adjusted, odds ratio (OR) for accumulated incidence of chronic bronchitis from 1993 to 2000 in the interventional villages were 0.80 (95%CI: 0.60-1.07) for men, 0.76 (95%CI: 0.45 1.28) in women. The OR for asthma was not significant in both men and women. CONCLUSIONS: Comprehensive interventions in community may improve knowledge of COPD prevention and living environments, decrease the smoking rate, cigarettes smoked per day, and incidence of chronic bronchitis, but have no significant effects on asthma. PMID- 15782502 TI - [Diagnosis and differential diagnosis of airway-centered interstitial fibrosis]. AB - OBJECTIVE: To describe a form of interstitial lung disease pathologically characterized by small airway-centered interstitial fibrosis (ACIF). METHODS: We analyzed the clinical, pulmonary functional, radiographic, and histologic characteristics of one ACIF case in Peking Union Medical College Hospital and reviewed 12 cases in literatures. RESULTS: Clinically, patients presented with chronic cough and progressive dyspnea. Pulmonary function tests showed restrictive ventilatory pattern. Bronchoalveolar lavage showed a mild increase in lymphocytes in most cases. Chest radiography revealed diffuse reticulonodular infiltrates, with thickening of the bronchial walls and surrounding fibrosis. The key finding in histopathology was a distinctive pattern of ACIF centered on membranous and respiratory bronchioles. CONCLUSIONS: ACIF is a disease that do not fit into any known category of interstitial lung disease. Whether it is a unique disease remains to be determined. PMID- 15782503 TI - [Magnetic resonance imaging of congenital vaginal anomalies]. AB - OBJECTIVE: To describe the magnetic resonance (MR) imaging manifestations in vaginal anomalies, and to discuss the utility and limit of MR imaging in diagnosis of this abnormality. METHODS: Nine female patients with congenital vaginal anomalies who required surgical intervention were studied. MR imaging and ultrasound of the pelvic was performed before surgery. MR imaging features were retrospectively analyzed correlating with surgery. RESULTS: The vaginal anomalies were categorized into three groups: (1) congenital absence of the vagina and uterus, i.e. Mayer-Rokitansky-Kuster-Hauser syndrome (MRKH syndrome) (n=3); (2) vaginal atresia (n=2); and (3) transverse vaginal septum (n=2). There was also one case of absence of vaginal with complicated urinogenital abnormality, and 1 case of obstructed hymen. CONCLUSIONS: MRI is a reliable method for evaluating vaginal anomalies. An accurate MRI examination can be helpful for surgical management. PMID- 15782504 TI - [Effect of age on the precision of dual X-ray absorptiometry measurement in middle-aged and elderly women]. AB - OBJECTIVE: To investigate whether age influence the precision of dual X-ray absorptiometry (DXA) measurement at the hip in middle-aged and elderly women. METHODS: Totally 90 women were randomly selected and divided into three age groups: 45-55 years, 56-65 years, and 66-75 years. Each age group contained 30 women. Each woman was scanned twice at the same day. Bone mineral density (BMD) values of femoral neck, ward's triangle, and trochanter were collected and grouped by calculating the root mean square (RMS). Precision errors were expressed as RMS (standard deviation, SD). RESULTS: For the femoral neck and trochanter, significant differences of SD of BMD existed among all age groups. For the ward's triangle, significant difference of BMD existed among all age groups except between the 45-55 group and 56-65 age group. CONCLUSION: Age can influence the precision of DXA measurement at the hip in middle-aged and elderly women. PMID- 15782505 TI - [Effect of neostigmine on atracurium-induced neuromuscular blockage in patients pretreated with magnesium sulphate]. AB - OBJECTIVE: To determine the effect of neostigmine on antagonizing atracurium induced neuromuscular blockage with sulfate magnesium pretreatment. METHODS: Forty patients who undertook elective gynecologic laparoscopic examinations and treatments under general anesthesia were randomized into four groups (group A, B, C, and D, group A paired with group C, and group B paired with group D). Before induction of general anesthesia, patients in group A and group C received MgSO4 30 mg/kg in saline intravenously within 5 min, while patients in group B and group D received the same volume of saline. Anesthesia was induced with fentanyl and propofol; subsequently tracheal intubation was performed with 0.5 mg/kg atracurium after stabilization of the electromyography recording, and neostigmine (0.02 mg/kg) and atropine (0.01 mg/kg) were infused in group C and group D when neuromuscular recovery (T1/T(C)) reached 10%. T1/T(C) changes after neostigmine infusion as well as haemodynamic changes and other responses during induction and neostigmine and atropine infusion were recorded. RESULTS: The neuromuscular recovery speed had no significant difference between group A and group B after the neuromuscular recovery reached 10%, but it was lower in group C than in group D (P < 0.05). Significant difference existed between group AC and group BD (P < 0.05). No haemodynamic changes and other responses were found during induction and neostigmine and atropine infusion. CONCLUSION: Neostigmine-induced neuromuscular recovery can be attenuated in patients pretreated with magnesium sulfate. PMID- 15782506 TI - [Recent advance in experimental study of cerebral ischemia treated by vascular endothelial growth factor]. AB - Vascular endothelial growth factor (VEGF) has been found to be the most powerful angiogenic factor. Studies have shown that cerebral ischemia and hypoxia stimulate the expressions of VEGF and its receptors in the brain, while exogenous VEGF promotes the formation of new blood vessels in the ischemic brain penumbra, and reduce the volume of cerebral infarction. The effect of VEGF on cerebral ischemia was previously explained the mechanism that VEGF had a specific mitogenetic roles in cerebral endothelial cells and thus promoted neovascularization; however recent evidence has shown that VEGF also has direct effects on neural and glial cells. Its multiple protection roles on central nervous system involve vascularization, neurogenesis, direct neurotrophic and neuroprotective effect, as well as antiapoptosis effect, especially when brain ischemia occurs. Further elucidation of these mechanisms on central nervous system may serve as a key procedure in understanding the main aspects of neural repair and neural protection, and develop effective therapeutic measures for intervention in stroke. PMID- 15782507 TI - [Ubiquitination-mediated degradation of epidermal growth factor receptor]. AB - After binding to its ligand, epidermal growth factor receptor (EGFR) dimerizes and is autophosphorylated. These events initiate the signal transduction process, which regulates a plethora of biologic activity. The duration and strength of these signals are controlled by many regulatory mechanisms, including downregulating activated EGFR primarily via endocytosis and ubiquitination dependent lysomal degradation. The interaction between EGFR and the ubiquitin ligase Cbl/adaptor protein CIN85, as well as ESCRT complex recruitment play important roles in the process of downregulating EGFR. Tumorigenesis results when the de-sensitization process of EGFR is halted by its own mutation or a mutation that abrogates Cbl E3 ligase activity. PMID- 15782508 TI - Explaining source of payment differences in U.S. cesarean rates: why do privately insured mothers receive more cesareans than mothers who are not privately insured? AB - The difference in the risk-adjusted cesarean rates of mothers who are and who are not privately insured is divided into components attributable to the following four factors: the practice style of the physician attending the birth, the closeness of the physician-mother relationship, individual nonclinical factors, and the direct influence of private insurance on the physician's treatment decision. Estimates from two expansive, complementary data sets indicate that the most of the differential is attributable to the first two factors, particularly the pairing of privately insured mothers with physicians who are inclined to perform cesareans. There is some evidence that these pairings are not incidental but the result of firm (and possibly consumer) choices. PMID- 15782509 TI - Improved allocation of HIV prevention resources: using information about prevention program production functions. AB - To allocate HIV prevention resources effectively, it is important to have information about the effectiveness of alternative prevention programs as a function of expenditure. We refer to this relationship as the "production function" for a prevention program. Few studies of HIV prevention programs have reported this relationship. This paper demonstrates the value of such information. We present a simple model for allocating HIV prevention resources, and apply the model to an illustrative HIV prevention resource allocation problem. We show that, without sufficient information about prevention program production functions, suboptimal decisions may be made. We show that epidemiologic data, such as estimates of HIV prevalence or incidence, may not provide enough information to support optimal allocation of HIV prevention resources. Our results suggest that good allocations can be obtained based on fairly basic information about prevention program production functions: an estimate of fixed cost plus a single estimate of cost and resulting risk reduction. We find that knowledge of production functions is most important when fixed cost is high and/or when the budget is a significantly constraining factor. We suggest that, at the minimum, future data collection on prevention program effectiveness should include fixed and variable cost estimates for the intervention when implemented at a "typical" level, along with a detailed description of the intervention and detailed description of costs by category. PMID- 15782510 TI - Engineering the economic value of two pediatric combination vaccines. AB - Combination vaccines for pediatric immunization have become an effective means to reduce the number of separate injections required to immunize children according to the United States Recommended National Childhood Immunization Schedule. This paper reports the results of using operations research methodologies to analyze the price and value of two pentavalent combination vaccines for pediatric immunization: diphtheria-tetanus-acellular pertussis, hepatitis B, inactivated polio (DTPa-HBV-IPV) and diphtheria-tetanus-acellular pertussis, Haemophilus influenzae type B, inactivated polio (DTPa-HIB-IPV). These two combination vaccines are analyzed both individually and head-to-head, as a function of the cost of administering (or avoiding) an injection and the number of doses of the vaccine required to be in the lowest overall cost vaccine formulary. The main contribution of the paper is to provide a methodology for analyzing the impact of combination vaccines on pediatric vaccine formularies. This analysis shows that the DTPa-HBV-IPV vaccine may provide a good value at the current federally negotiated price of 32.75 dollars for a wide spectrum of health-care environments, though the actual number of injections that it reduces may be fewer than the optimistic numbers claimed by its manufacturer. The analysis also shows that if the DTPa-HIB-IPV vaccine is approved by the Food and Drug Administration (FDA), then under current market prices, it may need to be priced below the sum of its vaccine component prices to favorably compete with the DTPa-HBV-IPV vaccine. PMID- 15782511 TI - Ambulatory care and orthopaedic capacity planning. AB - Ambulatory Care facilities (often referred to as diagnosis and treatment centres) separate the routine elective activity from the uncertainty of complex inpatient and emergency treatment. Only routine patients with predictable outcomes should be treated in Ambulatory Care. Hence the centre should be able to plan its activities effectively. This paper considers the consequences for the remaining elective inpatient bed and theatre requirements. Computer models are used to simulate many years of activity in an orthopaedic department at a typical District General hospital. The adoption of Ambulatory Care will increase the proportion of day case treatment but the reduction in the overall bed requirement will be relatively small (at most 10%). Separating the elective theatre activity into day case and inpatient sessions will tend to produce inpatient theatre sessions with a disproportionate number of longer procedures. This can reduce overall theatre utilisation by up to 15%, which implies the need for an increase of up to 18% in the number of theatre sessions if waiting times are to be maintained. PMID- 15782512 TI - Modeling patient flows using a queuing network with blocking. AB - The downsizing and closing of state mental health institutions in Philadelphia in the 1990's led to the development of a continuum care network of residential based services. Although the diversity of care settings increased, congestion in facilities caused many patients to unnecessarily spend extra days in intensive facilities. This study applies a queuing network system with blocking to analyze such congestion processes. "Blocking" denotes situations where patients are turned away from accommodations to which they are referred, and are thus forced to remain in their present facilities until space becomes available. Both mathematical and simulation results are presented and compared. Although queuing models have been used in numerous healthcare studies, the inclusion of blocking is still rare. We found that, in Philadelphia, the shortage of a particular type of facilities may have created "upstream blocking". Thus removal of such facility specific bottlenecks may be the most efficient way to reduce congestion in the system as a whole. PMID- 15782513 TI - A method for detecting runs of good and bad clinical outcomes on Variable Life Adjusted Display (VLAD) charts. AB - In recent years there has been a growing need for effective monitoring of clinical outcomes. Two techniques for continuous monitoring that have emerged almost simultaneously are the Variable Life-Adjusted Display (VLAD) and risk adjusted cumulative sum charts (CUSUM). The VLAD provides clinicians and management with an easily understandable overview of outcome history and is now in routine use in several hospitals. Although it can indicate runs of good and bad outcomes, unlike the CUSUM, it does not provide a quantitative means for assessing whether they merit investigation. This paper introduces a scheme for applying control limits from CUSUM charts onto the VLAD, thus enhancing its role as an effective monitoring tool. PMID- 15782514 TI - Health and economic impacts of an HIV intervention in out of treatment substance abusers: evidence from a dynamic model. AB - INTRODUCTION: A community-based intervention program found that the high-risk target population interacts with its surrounding community as a source of drugs and prostitution, creating a measure of co-dependence in the health status of each group. METHODOLOGY: The intervention collected extensive data on sexual and drug use practices in the target population. A dynamic compartment model estimates the epidemiological impact of the intervention, which serves as the basis for the economic assessment comparing intervention costs and lifetime HIV treatment costs. RESULTS: Approximately 2/3 of the new infections arise in the surrounding community. Intervention spillover benefits in the surrounding community are sufficient to make the intervention cost-saving in the first year- a savings of approximately 534,000 dollars. CONCLUSIONS: Conducting the intervention results in health benefits and cost-savings not only for the risk group, but for the entire community in which it resides. Quantifying the spillovers is vital to policymakers attempting to allocate scarce public health resources. PMID- 15782515 TI - Orofacial pain syndromes. PMID- 15782516 TI - Access to care. PMID- 15782517 TI - Practice-based research. PMID- 15782518 TI - The role of voice-activated technology in today's dental practice. AB - BACKGROUND: Voice-activated modules are available for most leading clinical software applications. This article briefly charts the evolution of this technology, describes how it is used in dental practices and discusses possible future developments. CONCLUSIONS: Improvements in speech recognition and microphone technology have helped voice-activated charting and clinical note dictation become more accurate, faster and easier to carry out than previously possible. As the technology continues to evolve and is applied more broadly in clinical settings, voice activation is likely to become a mainstream part of dental computer technology. PRACTICE IMPLICATIONS: Voice activation is a user friendly and cost-effective way to increase clinical productivity and maintain an innovative, up-to-date practice image. PMID- 15782519 TI - Using the modified Schirmer test to measure mouth dryness: a preliminary study. AB - BACKGROUND: Current methods of measuring saliva volume often are difficult to perform in a general dental or medical office setting. This study seeks to determine whether the Schirmer test for measuring eye dryness can be modified to measure mouth dryness. METHODS: The authors performed a modified Schirmer test (MST) on a control group composed of 41 healthy adult volunteers and a study group composed of 10 patients who had received head and neck radiation and 10 patients who had oral chronic graft-versus-host disease (CGVHD). The authors placed the strip on the floor of each subject's mouth and took readings at one minute, two minutes and three minutes. Fourteen control subjects also spat into a container over a five-minute interval, and the authors weighed the collected saliva. RESULTS: The mean reading for the control subjects at three minutes was 29.5 millimeters, while the mean reading for the test subjects was 6.9 mm (P < .00005). CONCLUSIONS: The results of the MST were able to distinguish between healthy adult volunteers and subjects who experienced profound xerostomia and hyposalivation. CLINICAL IMPLICATIONS: The MST as used in this preliminary study is an objective, inexpensive, easy-to-perform and well-tolerated test for measuring mouth dryness. It readily distinguishes between patients who are healthy and asymptomatic and those who experienced profound xerostomia and hyposalivation. PMID- 15782520 TI - Dental sealant longevity in a cohort of young U.S. naval personnel. AB - BACKGROUND: The U.S. Navy emphasizes caries prevention and encourages the placement of dental sealants on the caries-susceptible teeth of patients at risk of developing caries. The authors analyzed dental records to assess the longevity of dental sealants placed in naval personnel. METHODS: A cluster sample of dental records from 1,123 personnel who entered naval service in 1997 was drawn from eight Navy dental treatment facilities. The authors determined the number of sealants provided, the number of sealants that failed over the observation period (1997-2001), the dates of sealant failure and the longevity of sealants placed during and after recruit training. RESULTS: A total of 319 personnel received sealants during their first two years of service. The authors evaluated 1,467 sealed teeth. They followed the sealants for an average of 35 months. They noted 179 sealant failures in 102 subjects; 69 previously sealed teeth required sealant replacement, and 110 sealed teeth required restoration of the occlusal surface. Among those sealants that failed, the mean length of time from placement to failure was 26 months. Sealant failure rates were significantly higher among subjects at moderate risk or high risk of developing caries than among subjects at low risk. CONCLUSIONS: After an average of 35 months, 87.8 percent of the sealants placed in this population were retained and functional. Subjects who were at moderate or high risk of developing caries demonstrated significantly higher sealant failure rates than those at low risk of developing caries. CLINICAL IMPLICATIONS: Dental sealants can be retained successfully in adults. They should be considered a viable treatment alternative for adult patients who are susceptible to caries; however, patients at elevated risk of developing caries may require more frequent re-evaluation and maintenance to achieve maximum benefit. PMID- 15782521 TI - The impact of a brief tobacco-use cessation intervention in public health dental clinics. AB - BACKGROUND: Public health dental clinic patients use tobacco at disproportionately high rates. The purpose of this study was to evaluate a tobacco-use cessation program delivered via public health dental practitioners. METHODS: Two public health dental clinics participated in this quasiexperimental design study. First, all patients in one clinic who used tobacco (n = 178) received usual care. Next, the authors trained all practitioners to conduct a tobacco-use assessment and provide a brief cessation intervention. Subsequently, all patients in both clinics who used tobacco (N = 190) received the intervention. All enrolled patients had an income at or below the federal poverty level. The authors conducted follow-up assessments at six weeks and three and six months after enrollment. RESULTS: Differences in self-reported quitting by condition between participants in the two groups were significant across all endpoints. Patients in the intervention group were more likely to quit than those receiving usual care (15.5 versus 4.3 percent) and after 12 months (18.8 versus 4.6 percent). Controlling for enrollment differences between patients in the two groups (age, race/ethnicity, time to first cigarette after waking), the authors found that differences between groups were significant for quitting at three months (P < .05; odds ratio [OR] = 4.85; 95 percent confidence interval [CI] = 1.20, 19.60), and six months (P < . 01; OR = 5.25; 95 percent CI = 1.35, 20.36). CONCLUSIONS: The results of this study suggest the viability and effectiveness of delivering a tobacco intervention to low-income smokers via public dental practitioners. A randomized clinical trial is warranted. CLINICAL IMPLICATIONS: The potential reach of public health dental clinics is great. Because of the high percentage of tobacco-using patients in these clinics, the public health impact of a program such as the one reported here would be significant. PMID- 15782522 TI - Do procedural errors cause endodontic treatment failure? AB - BACKGROUND: This article reviews the effect of endodontic procedural errors, such as underfilling, overfilling, root perforations and separated instruments, on the outcome of endodontic therapy. TYPES OF STUDIES REVIEWED: Filling the root canal more than 2 millimeters from the radiographic apex (underfilling) or beyond the radiographic apex (overfilling), perforations of the root canal system and instrument separation are possible complications of endodontic therapy. Although these procedural errors may have different causes, they all may affect the outcome of treatment. RESULTS: Endodontic procedural errors are not the direct cause of treatment failure; rather, the presence of pathogens in the incompletely treated or untreated root canal system is the primary cause of periradicular pathosis. Procedural errors typically are due to several factors. Among them is a lack of understanding of the root canal anatomy, the principles of mechanical instrumentation and tissue wound healing. CLINICAL IMPLICATIONS: Procedural errors impede endodontic therapy, thus increasing the risk of treatment failure, especially in teeth with necrotic pulps and periradicular lesions. However, procedural errors often are preventable. PMID- 15782523 TI - Skeletal and dental changes with fixed slow maxillary expansion treatment: a systematic review. AB - BACKGROUND: The authors were not able to identify any systematic reviews or meta analysis on the dental or skeletal changes associated with slow maxillary expansion (SME). Therefore, they conducted a study to evaluate skeletal and dental changes after fixed SME treatment in orthodontic patients with constricted arches. METHODS: The authors included in their evaluation clinical trials that assessed skeletal and dental arch changes through measurements on dental casts or cephalometric radiographs. The authors did not consider trials involving surgical or other simultaneous treatment interventions during the active expansion period. RESULTS: With the help of a health sciences librarian, the authors searched the following electronic databases: PubMed, MEDLINE, MEDLINE In-Process and Other Non Indexed Citations, Evidence Based Medicine Reviews (Cochrane Database of Systematic Reviews, American College of Physicians Journal Club, Database of Abstracts of Reviews of Effects and Cochrane Central Register of Controlled Trials), EMBASE Excerpta Medica, Thomsen's ISI Web of Sciences and LILACS. Eight articles met the initial inclusion criteria. The authors found that a significant deficiency in the studies was the lack of a control group to factor out changes in the dental arch and skeletal structures associated with normal growth. CONCLUSIONS AND CLINICAL IMPLICATIONS: The authors found only a lower level of evidence. Therefore, they could make no strong conclusions on dental or skeletal changes that occurred after SME treatment. Clinicians need to rely on their clinical experience, experts' opinions and the presented limited evidence concerning SME treatments. PMID- 15782524 TI - Longevity of posterior tooth dental restorations. AB - Several forms of restorative techniques are used for posterior teeth. They vary significantly in cost and longevity. The following restorative concepts are the most commonly used: amalgam, resin-based composite, PFM, cast gold alloy restorations and all-ceramic restorations. I suggest that patients be informed about the potential longevity of restorative treatment for posterior teeth as they make decisions about treatment for their oral restorative needs. PMID- 15782525 TI - The ins and outs of dental insurance. AB - BACKGROUND: In the past three decades, dental insurance has become a permanent fixture in the delivery of dental services. Some would argue that this has not been good for dentists. However, none will dispute that it is here to stay and that dentists must make some very important decisions in their practices with regard to dental insurance. This article will explore some of those decisions and offer some guidance on how to decide wisely. CONCLUSIONS: Dental insurance now is a permanent fixture in the delivery of dental care. Each dentist must decide if participation with a dental insurance company is in his or her best interest. To make this decision, a dentist must have as much information as possible to evaluate each specific dental plan. If a dentist does decide to participate, the dentist must know the rules. PRACTICE IMPLICATIONS: Participation with a dental insurance company may influence the dentist-patient relationship. Dentists must be prepared to answer the question, "Why do I need this service if my dental insurance does not cover it?" Non-participation with a dental insurance company may result in a reduced number of new patients. PMID- 15782526 TI - Reimbursement for dental implants: dispelling some popular myths. AB - BACKGROUND AND OVERVIEW: Dental implants are being placed with increasing regularity. Many dentists do not take advantage of insurance coverages for this treatment because they are under the impression that no aspect of implant therapy is covered. This article discusses a number of insurance benefits that may be available to dental patients but not readily apparent to treating dentists and their staff members. CONCLUSIONS AND PRACTICE IMPLICATIONS: Some dentists and patients may assume incorrectly that a dental insurance plan does not reimburse for any implant therapy when, in fact, there may be some benefit available for at least a portion of the treatment. In addition, some dentists and patients may not consider implant therapy even when it is the preferred treatment option because of the assumed lack of reimbursement. Knowing that some reimbursement is available may make the difference in the patient's accepting the best treatment for his or her condition. Furthermore, since an increasing number of patients now make some contribution toward their dental insurance plan premiums, they deserve to know their options and to receive appropriate benefits. PMID- 15782527 TI - Practice progress. AB - This column is not based on wishful thinking about how things should be, but rather is a candid look at the reality of everyday practice. Prioritization is critical. Each dentist should select three to five major initiatives that he or she would like to achieve and prioritize these accordingly. The best approach for most practices is to deal with each of these only one or two at a time. Consider scoring each priority on a scale of one to 10 (one being the lowest, 10 the highest), gauging its overall importance, its potential effect on the practice and whether it helps achieve preset practice goals. PMID- 15782528 TI - Nationwide survey of work environment perceptions and dentists' salaries in community health centers. AB - BACKGROUND: Experienced private practitioners make up a significant proportion of dentists entering community health center (CHC) practices. The authors conducted a study to identify sources of dissatisfaction that affect the retention of these dentists and to determine current CHC dentist salaries. METHODS: CHC dentists nationwide were surveyed regarding salary and job satisfaction indicators. The authors mailed 569 surveys, and the response rate was 73.8 percent. The authors explored associations between job satisfaction indicators, salaries and dentists' intentions to leave the CHCs. RESULTS: Practitioners in private practice are the largest group of dentists recruited by CHCs (54.5 percent). However, 31.2 percent of currently employed dentists do not intend to remain in CHC dental practices. Salary was not associated significantly with the intention to leave. Years of experience, freedom of professional judgment, altruistic motivation, importance placed on loan repayment and amount of administrative time allowed were associated significantly with career change intentions. CONCLUSIONS: Periodic salary surveys can monitor factors associated with recruitment and retention of dentists in community and migrant health centers, and standardized exit surveys can identify factors causing dissatisfaction among dentists who leave. PRACTICE IMPLICATIONS: Employment opportunities in public nonprofit practices are increasing under current federal grant programs. However, unless job satisfaction issues are addressed adequately with dentists in social safety net programs, additional work force needs will not be met. PMID- 15782529 TI - An analysis of preventive dental visits by provider type, 1996. AB - BACKGROUND: Understanding preventive dental visit utilization patterns facilitates planning of the dental health services delivery system. The authors examine these patterns by analyzing the receipt of preventive dental services in the United States by type of dental provider. METHODS: The authors analyzed data from the 1996 Medical Expenditure Panel Survey (MEPS) for the U.S. community based population. They developed national estimates for the population with preventive dental visits by provider type, including the population with a preventive dental visit and mean number of preventive dental visits per person for socioeconomic and demographic categories. RESULTS: Respondents who are white, are older, are female, have dental insurance, are from higher income and education backgrounds, and reside in small metropolitan areas were more likely (P < .05) to receive preventive care from a dental hygienist than from a dentist. CONCLUSION: MEPS data showed that people's socioeconomic background and other demographic factors were associated with having a preventive dental visit with a dentist or dental hygienist. These factors also influence the per-person number of preventive visits by type of dental practitioner. These elements must be considered when planning for future dental work force needs. PRACTICE IMPLICATIONS: Estimating future dental work force needs through this analysis assists dentists in meeting patient demand and maximizing the productive output of all services rendered in their practices, including preventive services. PMID- 15782530 TI - The sanctity of peer review upheld. PMID- 15782531 TI - [Venezuelan Virology Network]. AB - In November 2004, sponsored by the World Bank, the Venezuelan Foundation of Science, Technology and Innovation (Fonacit) and the Venezuelan Institute of Scientific Research (IVIC), delegates from the different virology research groups of the country, met in Caracas-Venezuela, with the aim to establish the "Venezuelan Virology Network". The symposium entitled "Molecular biology applied to virus of health importance in Venezuela", was divided into three areas, including human and animals viruses related to public health: 1) Dengue, others arboviruses and Hemorrhagic Fevers; 2) diarrhea-related and others veterinary viruses and 3) Hepatitis, HIV and others sexually transmitted viruses. This symposium allowed the delegates to evaluate the current strengths, weaknesses and needs of the different laboratories, becoming evident the necessity of developing collaborative work between the groups that share the same interests or lines of research; and also their need to exchange technical resources, human and bibliographical material and consequently, avoiding the duplication of efforts and the unnecessary cost of resources. One of the main strengths of Venezuelan virology is the presence, in most laboratories, of researchers with studies of fourth level and multidisciplinary teams of work. We aspire to achieve the raised objectives in the event, to the benefit of our virology and even more important, of our people. PMID- 15782532 TI - [Gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha) and interleukins 2, 4 and 6 (IL-2, IL-4, IL-6) in cervical-uterine cells of intraepithelial neoplasia: a preliminary report]. AB - The purpose of this work was to determine the expression of type Th1 cytokines: IL-2 and IFNgamma, and Th2: IL-4 and IL-6, as well as TNF-alpha in patients with precancerous lesions of the uterine cervix and their relationship with the human papiloma virus (HPV). 30 patients with precancerous lesions (NIC 1: 70%, NIC 2: 16.7% and NIC 3: 1.3%) and 9 normal controls were studied. A clinical history, gynecological evaluation, cytology and an uterine biopsy were carried out in each patient and control. PCR was used for the diagnosis of HPV. IFN-gamma expression (positive cells/field) was increased in patients with NIC (5.06 +/- 4.7 vs 0 in the control group; p < 0.05). TNFa was a little higher in pathologycal tissues than in the controls (5.23 +/- 3.63 vs 1.55 +/- 2.65; p < 0.05). IL-2 was higher in pathologycal cases than in the controls (8.73 +/- 5.23 vs 0.33 +/- 1, p < 0.05). IL-4 were expressed in both, patients and controls (6.53 +/- 5.23 vs 5.77 +/- 7.32). IL-6 was also higher in patients (4.63 +/- 3.34 vs 0.77 +/- 2.33; p < 0.05). When the HPV status was considered, only IFN-gamma (p < 0.05) and IL-2 (p < 0.05) were significantly higher in HPV positive patients (n = 4) compared to controls. When HPV+ patients were compared with HPV- patients, only IFNgamma was significant (11.5 +/- 5 vs 4.07 +/- 3.8; p < 0.05). In conclusion, Type Th1 immune response prevails in patients with precancerous lesions, whether they are HPV positive or not. PMID- 15782533 TI - [Oropharyngeal carriers in Venezuelan children in two child care settings, vaccinated or not with Haemophilus influenzae type b tetanus toxoid conjugate vaccine (PRP-T)]. AB - The objective was to estimate the oropharyngeal carrier rate of Haemophilus influenzae type b (Hib) and non typeable (NTHi) in unvaccinated and vaccinated children, with PRP/T vaccine, both cared at home and attending day care centers. Healthy children of either sex, younger of 7 years, excluding those that received antibiotics one month prior to the inclusion, were recruited from control clinics for healthy babies, and assigned to Group I fully immunized and Group II unimmunized. Both, children cared at home and attending day care centers were included. Parents provided an informed consent. Specimens were obtained from the oropharynx for culture, identification and serotyped by slide agglutination (Phadebact Haemophilus test). P values were obtained with Chi-square, Fisher's and Z test. Odds ratio was obtained when appropriated. Results were as following: a total of 320 children were enrolled, 99 in group I (49 cared at home, 50 attending day care) and 221 in group II (94 cared at home, 127 attending day care) the percentage of carriers was 22.8% (17.8% NTHi and 5% Hib). The carriage rate of NTHi was similar in groups I and II cared at home (18% vs 19%), and for those attending day care were 16% vs 17%. Carriage rate of Hib in children attending day care of group II was 12% vs 1% of those cared at home (p = 0.002). None of the children in group I was colonized by Hib. The Hib carrier rate was more similar to that of industrialized countries in the pre vaccine era in children cared at home, however, attendance to a day care center significantly increases its risk. The PRP/T vaccine appears to prevent the carrier state in both care settings. PMID- 15782534 TI - [CXCR-4 AND CCR-5 expression in normal term human placenta]. AB - The maternal-fetal interphase has an active Immunitary System (IS) whose mediators -cells, cytokines and chemokines- coordinately act to favour pregnancy normal development. It is not known exactly which of those mediators are present in each placental cellular stratus and what the physiological or potentially pathologic consequences derived from their presence can be. It is known that chemokines recruit cells with regulatory activity towards the deciduous and some of their receptors are coreceptors to infectious agents like HIV, making research of chemokines expression and their receptors in the maternal-fetal interphase of great interest in recent years. In the present study, the CXCR-4 and CCR-5 expression was investigated in 8 samples of normal human placenta obtained from term pregnancies, with low obstetric risk, by using Immunocitochemical techniques (Biotin-Avidin-Peroxidase). The most relevant finding in this study was the demonstration that CXCR-4 and CCR-5 differential expression in trophoblast, stroma and endothelium represents, as far as we know, the first report of the presence of these receptors in all layers of placental tissue. These results help to broaden the knowledge about the expression of chemokines receptors -that act as main coreceptors in the HIV infection- in the maternal-fetal interphase, and this can be a contribution to be taken into account in the vertical transmission study of this infectious agent. PMID- 15782535 TI - [Mycoses at Hospital Universitario "Ruiz y Paez", Ciudad Bolivar, Venezuela, 2002]. AB - To estimate the prevalence of mycoses in the Universitary Hospital "Ruiz y Paez" (Bolivar State, Venezuela) during 2002, a retrospective study was carried out. Four hundred and fifty six mycoses were diagnosed in 250,956 patients. The most frequent mycoses, the cutaneous form (94.5%), were produced by dermatophytes, Malassezia furfur and Candida spp. in 90%. Deep mycoses were detected in 4.7% and consisted of histoplasmosis (2.6%), paracoccidioidomycosis (1.7%) and cryptococcosis (0.4%). Several patients with deep mycoses presented HIV infection, tuberculosis and hematological disorders (neoplasias among them). The frequency of mycoses in the general population that attend the Hospital Universitario "Ruiz y Paez" is low, superficial mycoses are predominant, and some conditions seem to predispose to deep mycoses. PMID- 15782536 TI - [Evaluation of infection by Helicobacter pylori in HIV positive patients trough enzyme immunoassay and specific amplification of DNA]. AB - The objective of this work was to assess the effectiveness of detection of specific antibodies anti-Helicobacter pylori (H. pylori) by ELISA and amplification of specific DNA by polimerase chain reaction (PCR) as diagnostic methods of infection of H. pylori in HIV positive patients. Twenty two patients with HIV infection were studied, with ages between 26 to 35 years, 17 masculine, 55% with gastrointestinal symptoms, controlled in the Unit of Immunology, CHET. Inclusion approaches: older than 18 years, with confirmed diagnosis of HIV infection (ELISA and WB), lymphocyte subpopulation and good general conditions. Consent in writing was obtained. Exclusion approaches: previous diagnosis of H. pylori infection or treatment with antibiotics in the three previous months to their inclusion. The quantification of IgG anti H. pylori was carried out by Enzyme Immunoassay methods (ELISA). Biopsy of gastric mucosa was obtained by superior endoscopic study. The amplification of DNA for H. pylori was performed by PCR (Wizard SV Genomik and PCR Ready-Promega). In the statistical analysis was used the test of Fisher, with a level of significance of 5% (0.05). In 15 patients of the total group, antibodies anti H. pylori were confirmed, without statistical association with the presence or not of digestives symptoms, neither with the number of lymphocytes CD4 + in peripheral blood. Also 15 patients were positives by PCR for H. pylori DNA, 73.3% of them presented levels of CD4+ above 200 cells. There was not statistical association between the positivity of this method and levels of lymphocytes CD4+. In 12 of the 15 patients with positive results by PCR, antibodies anti H. pylori were evidenced, and among the 7 patients with negative serology to H. pylori, PCR was positive in three of them. In conclusion, serology is an effective method for the diagnose of H. pylori infection in VIH+ patients, but its negativity doesn't discard the infection for this bacillus. PMID- 15782537 TI - [Vitamin B12, folic acid and mental function in the elderly]. AB - Elderly people is a vulnerable population group to specific nutrient deficiencies as vitamin B12 and folic acid, which are closely related to mental functions deterioration, especially of cognitive functions. This study was aimed to measure B12 vitamin and folic acid indicators and to establish relationships to mental function. 53 elderly, older than 60 years, living in a geriatric home were assessed. The dietary intake was evaluated by the direct weighed method, serum B12 vitamin and folic acid by radioimmunoanalysis and mental function by Foltein's mini-mental test. Dietary intake for Vit B12 was adequate and deficient for folic acid while serum levels were within normal range. Vitamin B12 levels were at marginal or deficiency values in 26,4% of the elderly and folic acid deficiency was present in 43.4%. 49% of the elderly had mental function alterations and B12 vitamin levels were significantly lower in this group. A positive association between age and mental function (elderly below 80 years had lower risk of mental impairment) and between serum B12 and mental function were found. Elderly were at risk of deficiency for both vitamins and age and mental function were associated to this risk. Further evaluation including other nutrients should be performed. PMID- 15782538 TI - [Superficial mycoses: comparative study between type 2 diabetic patients and a non-diabetic control group]. AB - Superficial mycoses are considered to affect more frequently patients with type 2 diabetes mellitus (DM-2), specially onychomycosis and Tinea pedis. The purpose of this study was to compare the dermatophytoses, candidiasis and Pitiriasis versicolor frequency between 40 patients with DM-2 and 40 healthy persons of either sex, 40 years old or more. Clinical, metabolic, mycologic and inmunologic studies against Candida albicans, were carried out. Both diabetics 75% (30/40) and controls 65% (26/40) presented a high frequency of superficial mycoses (no significant difference p = 0.329). Pitiriasis versicolor was not detected in diabetic patients. They presented Tinea unguium, concomitant with Tinea pedis, with a higher frequency. The predominant dermatophyte was Trichophyton rubrum 18/23 (78%) in diabetics and 8/16 (50%) in non diabetics. Candida was isolated as commensal from oral mucous: 23/40 (58%) in diabetics and 21/40 (52%) in non diabetics (serotipo A was the more frequent), and from onychomycosis: 11/40 (28%) in diabetics and 12/40 (30%) in non diabetics. The immunological response was the same in both groups: celular 100%, humoral 20%. No statistical correlation among superficial mycoses, blood glucose level, glycosylated hemoglobin values or the time suffering the disease was observed. The high susceptibility to dermatophytes and Candida sp. infection showed to be associated with age and no with the diabetic type 2 condition in those patients. PMID- 15782539 TI - [Severe acute respiratory syndrome (SARS). Lesson and challenges]. AB - After four months of silence before a new incursion of severe acute respiratory syndrome virus (SARS), world wide investigators specially from Asia had have to face a coronavirus capable of great spreading and inducing high morbidity. The source and the capacity of this virus to associate with other opportunist microorganisms to induce progression of the disease, remain unclear. The comparison of the genomic sequence of different strains has generated discrepancies as for if the SARS epidemic arose as an unique outbreak or it was produced for more than one genotype. However, this fact is secondary to the imminent threat that was controlled in August of 2003, after affecting 8.422 individuals, but before which we should be remain alert, since it is clear that the end of the SARS is not yet here; and that the appearance of a single case can mean the beginning of a new wave, joining the outbreak of other diseases such as the avian flu. In this regard, it is very important to know the epidemic bases of morbid-mortality of SARS and to keep a world wide surveillance in order to detect possible further cases. PMID- 15782540 TI - Antibodies to tumor necrosis factor-alpha in the treatment of Crohn's disease. AB - Crohn's disease is characterized by acute and chronic intestinal inflammation, and can involve any part of the gastrointestinal tract. The most frequently involved areas of the intestine (the terminal ileum and colon) have higher bacterial loads than the rest of the intestine, suggesting a role for a dysregulated immune response to luminal bacteria. Research into the pathogenesis of Crohn's disease using animal models supports a role for Th1-mediated immune responses, and inhibition of the generation of a Th1 response is known to prevent disease. Based on these observations, antitumor necrosis factor-alpha (anti TNFalpha) therapies have been used to treat patients with Crohn's disease and more recently with ulcerative colitis. Certain anti-TNFalpha therapies, such as infliximab, have resulted in dramatic clinical responses in patients with Crohn's disease. Other therapies such as etanercept, however, have not been effective. In this review we will discuss the different strategies that have been employed to inhibit TNFalpha and their relative merits. We will also address factors that predict response to therapy such as concurrent immunomodulators, high C-reactive protein expression, and polymorphisms in the Fcgamma receptor. PMID- 15782541 TI - Immunotherapy of rheumatoid arthritis: past, present and future. AB - Rheumatoid arthritis (RA) is a chronic autoimmune disease, characterized by inflammation of the synovial lining of joints, and the destruction of cartilage and bone. Seminal studies demonstrating that pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNFalpha), are expressed in RA, has resulted in the approval of anti-TNFalpha biological therapies for its treatment. Although groundbreaking in themselves, these studies have also paved the way for further research to determine whether the targeting of other cytokines and immune pathways might aid in development of the next generation of drugs for the treatment of RA. PMID- 15782542 TI - Effective tumor targeting: strategies for the delivery of Armed Antibodies. AB - It has long been realized that the presence of tumor-associated antigens offers an excellent opportunity for targeted cancer therapy and hence an improved clinical benefit for cancer patients. Advances in the field of antibody engineering as well as the characterization of toxins, such as diphtheria toxin and Pseudomonas exotoxin A, have enabled the routine construction of recombinant immunotoxins, which we have termed Armed Antibodies. The selective toxicity and mechanism of action of these molecules could potentially provide an excellent clinical alternative to conventional anticancer agents which have many unacceptable side effects. Although considerable clinical success has been achieved using immunotoxin therapy, particularly for B-cell malignancies, the treatment of solid tumors remains highly challenging. To successfully treat solid tumors that are not amenable to local therapy, immunotoxins must be designed to permit repeat systemic administration. This review outlines some of the strategies currently being employed in the design of the Viventia Biotech Inc Armed Antibodies to minimize the development of immunogenicity and to remove the potential for toxicity in non-target tissues. PMID- 15782543 TI - The yin and yang of cancer therapeutics. AB - In this review, a potential theoretical framework for cancer therapeutics is presented, based on two concepts. The first concept considers cancer as a 'disease process' involving key regulatory pathways, and is discussed along with recent evidence and topics of special interest such as the combination of molecular diagnostics with developmental therapeutics. The second concept is the binary state concept of 'active/inactive' that seeks more relevant targets within the global molecular matrix of any given cancer. Here, these concepts are considered, along with the growing body of evidence that supports them. The 'binary' concept may facilitate both target selection and target validation. PMID- 15782544 TI - Recent advances in hepatic gene transfer: more efficacy and less immunogenicity. AB - Systemic enzyme deficiencies and lysosomal storage disorders are excellent targets for the treatment of genetic disease by in vivo gene transfer. To enable efficient delivery of therapeutic gene products into the systemic circulation, gene transfer to the liver has been extensively pursued. Hepatocytes are capable of overexpressing biologically active enzymes, such as coagulation factors (in the treatment of hemophilia) and lysosomal enzymes, and efficiently secrete these proteins into the blood stream. Sustained therapeutic expression and correction of rodent and canine animal models of human disease has been reported for a number of genetic disorders. This has been possible because of recent advances in vector development and optimization of their delivery. Viral vectors, in particular adeno-associated viral vectors and retroviral vectors, have yielded remarkable successes in the treatment of dogs with hemophilia or mucopolysaccharidosis. Such studies in large animals represent an important intermediate step toward clinical application, which has now been initiated in the case of hemophilia. Equally importantly, hepatic gene transfer has been demonstrated to induce immune tolerance to therapeutic transgene products. PMID- 15782545 TI - The search for novel drug leads for predominately antitumor therapies by utilizing mother nature's pharmacophoric libraries. AB - Recent advances in the use of natural products and compounds derivedfrom natural products as agents for the treatment of cancer and other diseases are reviewed. The antitumor agents discussed include tubulin interactive agents, inhibitors of topoisomerases I and II, caspase activators, proteasome inhibitors, histone deacetylase inhibitors, heat shock protein 90 inhibitors, protein kinase inhibitors, and inhibitors of hypoxia inducible factor 1. It can be concluded that, despite a perceived lack of popularity in recent years, the use of natural product scaffolds in searching for drug leads is still being practiced. PMID- 15782546 TI - Novel natural compounds obtained by genome-based screening and genetic engineering. AB - Despite the fact that drugs derived from natural products have revolutionized medicine in the past, they are currently going through a phase of reduced interest in drug discovery. At the same time, however, there is an urgent medical need for new drugs, since development pipelines are drying up and resistance to antibiotics and other chemotherapeutic agents is becoming an increasingly frequent problem. The development and recent progress of new technologies, such as genetic engineering and screening, offer a unique opportunity to re-establish natural products as drug leads. Examples of recent advances in the application of these technologies to the discovery and development of important novel drugs are discussed in this review. PMID- 15782547 TI - Protein microarrays: applications and future challenges. AB - Within the last decade protein microarray technology has been successfully applied for the simultaneous identification, quantification and functional analysis of proteins in basic and applied proteome research. These miniaturized and parallelized assay systems have the potential to replace state-of-the-art singleplex analysis systems. However, prior to their general application in robust, reliable, routine and high-throughput applications it is mandatory that they demonstrate robustness, sensitivity, automation and appropriate pricing. In this review, the current state of protein microarray technology will be summarized. Recent applications for the simultaneous determination of a variety of parameters using only minute amounts of sample will be described and future challenges of this cutting-edge technology will be discussed. PMID- 15782548 TI - From mouse genetics to human therapeutics. AB - This review examines how and where genomic and genetic research will impact pharmaceutical research and development, and emphasizes how mouse genomics and genetics can improve the understanding of human disease pathobiology and drug metabolism, and identify new targets for therapeutic intervention. Although important discoveries can be made from mouse genetic analysis, its utility has been limited by the high cost and long time lines required for such research. A recently developed computational method that markedly accelerates the pace of genetic discovery and reduces its cost is also described. PMID- 15782549 TI - Modulation of gene expression by siRNA in hematopoietic cells. AB - RNA interference (RNAi) has been established as a powerful tool for identifying gene function in many biological processes and can be used for genome-wide functional genetic screens in mammalian cells. For such purposes, expression cassettes encoding RNAi triggers can be efficiently introduced into the host cell genome utilizing viral vector systems, resulting in long-term silencing of target gene expression. Transient gene silencing can also be induced by exogenous delivery of suitable RNAi triggers to target cells. However, similarly to other reverse genetic tools, there are technical challenges and limitations associated with RNAi, some of which are specific to hematopoietic cells. In this review we discuss the rational design of effective RNAi triggers, different approaches for their efficient delivery, and the value of RNAi both as a potential therapeutic strategy and as a tool for functional genomics and target validation in hematopoietic cells. PMID- 15782550 TI - PPARs: molecular targets in the pharmacogenomics era. AB - The recent global increase in prevalence of diseases like obesity, type 2 diabetes and hypertension in westernized societies is unfortunately not paralleled by our full understanding of the causative mechanisms. It is now firmly established that the interacting genetic and environmental (diet, smoking) components together determine the development and severity of the particular condition, which makes detailed dissection of such complex traits even more complicated. In effect, there is an unmet urgent need for molecular targets so we can directly modulate the causative factors and devise effective preventive and therapeutic algorithms. Among the most promising molecular targets for treatment of metabolic syndrome-related conditions identified so far, the group of three lipid-sensors, the peroxisome proliferator-activated receptors (PPARs) clearly stands out. The review focuses on pharmacogenetic aspects of recent developments in PPAR biology. PMID- 15782551 TI - ANCA-associated renal vasculitis--epidemiology, diagnostics and treatment. AB - The pauciimmune small-vessel vasculitides are multisystem diseases with frequent renal involvement. They are strongly associated with the presence of anti neutrophil cytoplasmic antibodies (ANCA). In this review we have focused on the ethiopathogenesis and the role of ANCA, clinical presentation and histopathologic findings of different ANCA - associated vasculitides (AAV). Current treatment strategies and the overall and renal outcome of patients with AAV are also discussed. PMID- 15782552 TI - New immunosuppressive agents in chronic progressive glomerulopathies: an update. AB - The etiology of chronic glomerulopathies is not yet clear, however the impairment of immune system is supposed to play a decisive role. Thus, immunosuppressants are often used to slow the activity and/or progression of the disease. Usually the patients are treated either by corticosteroids alone or by the combination of corticosteroids with other immunosuppressive agents. Recently, immunosuppressants successfully applied in the transplantation medicine have been also tested in the therapy of chronic glomerulopathies. Cyclosporine A, a potent calcineurin inhibitor, has been used as a second line therapy. The place of CsA in lupus nephritis and especially in IgA nephropathy is not defined so well as it is in idiopathic nephrotic syndrome. The newer calcineurin inhibitor tacrolimus is already widely used in the transplantation medicine while its effectiveness in clinical nephrology has to be tested. Mycophenolate mofetil in monotherapy or with concomitant low-dose steroids can be regarded as an alternative therapeutical approach in case of standard regimens' failure. Sirolimus just entered clinical transplant medicine and its role in slowing the progression of chronic glomerulopathies is not yet clear. PMID- 15782553 TI - Surgical contribution to the management of primary hyperparathyroidism. AB - Surgical management of primary hyperparathyroidism is a very effective method. The target is to cure primary hyperparathyroidism and to reach normal calcium levels. This results in an improvement of health condition and resolution or at least moderation of symptoms. Complications are infrequent and mortality is very low. Surgical management is definite, safe and effective. Authors of this article address the diagnosis of primary hyperparathyroidism, clarify bone, metabolic and biochemical syndromes and present series of 151 patients that have been operated on at the 3rd Department of Surgery of the Motol University Hospital, Prague, with the diagnosis of primary hyperparathyroidism. The survey is focused on the primary hyperparathyroidism concomitant diseases and on the possible sequelae in the postoperative period. PMID- 15782554 TI - Effects of intracarotid injection of methylprednisolone on cellular oedema after osmotic opening of the blood-brain barrier in rats. AB - In our work we studied methylprednisolone (MP) for its effects on the permeability of cytoplasmatic membranes of neuronal populations in the rat. We used a standard model of cellular oedema induced by water intoxication, applying MP selectively into the internal carotid (ICA) after opening the blood-brain barrier (BBB) with mannitol. The results were assessed under fluorescence microscopy in keeping with the Intracellular Distribution Index of Evans Blue (IDI) in the neocortical field (Cortex) and in hippocampal areas CA1, CA3 and GD. Evans blue (EB) was applied similarly as MP Three different experiments were carried out. In experiment 1--EB alone and no MP was applied. In experiment 2- 5.4 mg/kg MP and EB were applied. In experiment 3--54 mg/kg MP and EB were applied. In experiment 1 the IDI values were high (>1), indicating the presence of large quantities of EB in the cells. In experiments 2 and 3 the IDI values were low (<1), indicating more EB outside than inside cells. IDI differences between experiments 2 and 1 and experiments 3 and 1 were statistically significant (p<0.05). This morphological evidence sufficiently proved the possibility to restore the cell membrane integrity by means of MP administration. PMID- 15782555 TI - Hypoxia-induced long-term increase of dopamine and tyrosine hydroxylase mRNA levels. AB - The aim of the present study was to determine hypoxia-induced changes in the long term expression of tyrosine hydroxylase (TH) mRNA and the steady-state dopamine (DA) levels in rat mesencephalic cell cultures. The cultures were exposed to hypoxia during the early developmental period, and DA content and TH mRNA expression were determined on day in vitro (DIV) 14. Hypoxic exposure of 5-day old cultures resulted in increased DA (control 89.9+/-8.9, hypoxia 135.8+/-23.7 pg/microg protein) and TH mRNA (control 37.3+/-4.7, hypoxia 143.1+/-49.4 pg/microg RNA) levels. To analyze the involvement of hypoxia-inducible factor-1 (HIF-1) in these changes, we studied its activation using reporter gene. Hypoxia caused a 3-fold increase in HIF-1 activity. Our data suggest that hypoxia/ischemia during the putative critical developmental period of neurons may determine the tyrosine hydroxylase gene expression and, consequently, the development of the dopaminergic system. PMID- 15782556 TI - Markers of inflammation in preeclampsia. AB - Advanced oxidation protein products (AOPP) represent terminal products of proteins exposure to free radicals. The aim of this study was to estimate the serum AOPP levels in preeclamptic patients together with ultrasensitive C reactive protein and anticardiolipin antibodies (ACA) IgG and IgM. 21 women in the third trimester of pregnancy were included in the study--10 women with preeclampsia and 11 women with normal outcome of pregnancy. AOPP levels in preeclampsia were higher than those in normal pregnant women in the third trimester, but not statistically significantly. The comparison with AOPP levels in non-pregnant women has shown a significant increase (P<0.0001). CRP in preeclampsia was significantly increased in comparison with third trimester levels in normal pregnancy (P<0.001) as well as with non-pregnant women (P<0.0001). In preeclampsia, the ACA IgG levels were even significantly lower than in normal pregnant women in the same gestation age, but significantly higher than in non-pregnant women (P<0.001). No difference was found in ACA IgM in preeclampsia and normal third trimester pregnancy and non-pregnant women. A statistically significant negative correlation was found between AOPP and ACA IgG (r= - 0.708, P<0.05). The results indicate enhanced oxidative and inflammatory reaction of maternal organism to pregnancy, which is more pronounced in preeclampsia than in uncomplicated pregnancy. PMID- 15782557 TI - Thrombelastography monitoring of platelet substitution therapy and rFVIIa administration in haemato-oncological patients with severe thrombocytopenia. AB - Thrombocytopenic patients refractory to platelet concentrates (PC) could be treated during bleeding episodes with the recombinant activated FVII (rFVIIa). However, monitoring of administration of the rFVIIa or a response to platelet substitution therapy in thrombocytopenia patients is not well documented so far. Using of whole blood ROTEG analysis we monitored the changes in haemostatic parameters following in vivo platelet concentrate administration compared to ex vivo rFVIIa administration in patients with a severe to mild thrombocytopenia secondary to haemato-oncological disease. We use non-activated thrombelastography (NATEG) and a mild intrinsic activation thrombelastography (INTEG). NATEG analysis was sufficiently sensitive to monitor changes following PC and rFVIIa administration. Both, platelet infusion and rFVIIa treatment induced significant shortening of clotting time (CT) and clot formation time (CFT) parameters (p<0.05). When we compared the effect of platelet vs. rFVIIa treated whole blood by NATEG analysis we did not found any significant difference. Analysis with INTEG system was less sensitive and changes in CT and CFT were not significant. The monitoring with thrombelastography could enable efficient application of platelet concentrate and furthermore the using of rFVIIa as an alternative treatment of patients refractory to platelet infusion or with allergic reactions. PMID- 15782558 TI - Microphthalmia transcription factor: a specific marker for malignant melanoma. AB - The transcription factor microphthalmia (MITF) is required for the formation of normal melanocytes during embryonic development and for the expression of pigment cell-specific markers, which are the downstream transcriptional targets of MITF. It also seems to be crucial for the survival of malignant melanocytes. The special interest of this review is the possible utility of MITF as a marker of malignant melanoma. Melanocyte-specific isoform of MITF appears to be a unique molecule in the differential diagnosis of melanocytic tumors. PMID- 15782559 TI - Plasticity of the brain in neuroontogenesis. AB - Plasticity is a specific endowment of the nervous system to develop, to react or to adjust to the internal and external 'environmental changes, both in the physiological and pathological conditions. Cumulative evidence has revealed the dynamism of the nervous system, based on the balance between the rigidity and plasticity. Different aspects of neuroplasticity can employ common general cellular mechanism. Effects of plasticity can be either positive or negative changes during the development (evolutional plasticity), after the short-term exposition (reactive plasticity), after the long-term or permanent stimuli (adaptational plasticity), and during functional or structural recovery of the damaged neuronal circuits (reparation plasticity). Manifestations of plasticity have probably the same basis, irrespective of a cause, which triggered them, or the brain region where they were accomplished. Activity of neuroplastic processes appears to be especially high in the immature nervous tissue. PMID- 15782560 TI - Microsatellite markers in breast cancer studies. AB - Microsatellites are tandem repeats of simple polymorphic sequences randomly distributed in non-coding regions of DNA. They can be used in cancer genetics and indirect cancer diagnosis and can help unraveling the genetic basis of tumor formation and progression of cancer. Breast cancer is a complex disease in which numerous genetic alterations occur. The knowledge of specific genetic changes and their biological consequences is critical to an understanding of breast cancer tumorigenesis, screening and treatment of patients. Microsatellites can undergo two events during tumor progression. Loss of heterozygosity indicates absence of one allele in a given locus, which is associated with the loss of a corresponding genes. Microsatellite instability reflects replication errors induced by defective function of mismatch repair genes and is demonstrated with the appearance of novel, noninherited alleles in tumor cells and represents a specific pathway of tumor development. Both events serve as prognostic markers, which can be correlated with clinicopathological features and can help exploring breast cancer formation. PMID- 15782561 TI - Effect of selenium pre-treatment on evoked cortical afterdischarges in young rats. AB - Prenatal or perinatal hypoxia is among the most frequent pathogenic factors of encephalopaties. It can induce wide-range of morphological, biochemical, energetic and functional alterations. Accordingly we tested changes of excitability of the sensorimotor cortex in 12, 25 and 35-day-old rats exposed and not exposed to short-term (1 hour) hypobaric and normobaric hypoxia. We studied whether sodium selenate (0,26 mg/kg b.w.) can influence character and/or intensity of evoked cortical afterdischarges or if a pre-treatment by selenium changes effects of hypoxia on such seizures. According to the results we can conclude that in our experiment arrangement sodium selenate only partially alters duration of evoked cortical afterdischarges. PMID- 15782562 TI - Electrocardiographic body surface maps (BSM) in patients with ischemic heart disease examined by coronary angiography. AB - ECG body surface maps (BSM) is one of the noninvasive methods for the detection of ischemic heart disease. In the present work we registered the BSM in 25 patients, 18 men, mean age 56.8 (31-83) years, 7 women, mean age 58.7 (43-72) years with coronary artery disease and in 23 healthy persons, 17 men, mean age 55 (46-60) years, 6 women, mean age 57 (42-70) years. Using diagnostic system Cardiag we measured 32 parameters of heart electric field (ECG, VCG, isopotential, isointegral, and isoarea maps). The results of BSM examination were compared with the results of coronary angiography (CAG) and other noninvasive methods. Twenty-four patients were BSM positive, 5 of them with negative CAG. In one case both BSM and CAG were negative. None of persons with positive CAG were BSM negative. The values of positive BSM in patients with negative CAG approached more to the values of positive CAG persons than to controls. Despite of small number of persons examined in our study one can conclude that the BSM method detects the damage of myocardium by another way than the CAG. PMID- 15782563 TI - 105th birth anniversary of professor Frantisek Por, M.D. AB - Professor Frantisek Por, M.D., was one of the most remarkable physicians in Czechoslovakia. He graduated at the German Medical Faculty of Charles University in Prague, in 1926. He was a founder of the First Internal Clinic of the Medical Faculty of P J. Safarik University and of the Faculty Hospital in Kosice. Prof. F. Por, M.D., was the Head of the First Internal Clinic from 1948 until 1971. During his active professional life he educated 11 associate professors and 3 full professors. He was also a founder of Eastern Slovakian Medical Meetings in Novy Smokovec, the High Tatras, in 1961. The "Memorial Meeting of Professor F. Por, M.D." has been organized by the Medical Society in Kosice since 1994. The last one was held in the Faculty Hospital of L. Pasteur in Kosice, on April 28, 2003. PMID- 15782564 TI - Charting the future of the Japanese Association for Thoracic Surgery: where to from here? PMID- 15782565 TI - Discrepancy between Doppler and catheter measurements of pressure gradients across small-size prosthetic valve. AB - OBJECTIVES: The exact role of pressure gradient across the prosthetic valve estimated from Doppler flow velocity remains controversial. This in-vivo study was designed to assess the actual discrepancy between Doppler and catheter measurements of the pressure gradients for small bileaflet prosthetic valves in the aortic position. METHODS: Bileaflet prosthetic valves (19 mm-ATS) were implanted into the aortic position in pigs, and pressure gradients across the valves were examined by volume loading under right heart bypass. The pressure gradient obtained by catheter was defined as the conventional peak-to-peak gradient between the left ventricle and aorta. The peak Doppler gradients were calculated from the maximal instantaneous Doppler velocity with the ultrasound probe positioned on the diaphragm at the level of the cardiac apex. RESULTS: There were strong correlations between pressure gradients and cardiac output. The Doppler gradient was constantly higher than the catheter values, and the resultant discrepancy between Doppler and catheter measurements was directly dependent on cardiac output (y=9.9x+0.6, r2=0.55). For cardiac output > or = 5.0 L/min, the difference between Doppler and catheter gradients reached 40 mmHg, and maximum differences of up to 80 mmHg were observed. CONCLUSIONS: In view of the presence of striking overestimation of catheter gradient by Doppler measurement, Doppler ultrasound should be used cautiously to assess small-size bileaflet prosthetic valve function with consideration of the patient's hemodynamic state. PMID- 15782566 TI - Leaflet folding plasty for mitral valve repair: technical application and early outcome. AB - OBJECTIVES: Leaflet folding plasty was introduced for avoiding systolic anterior motion and coronary artery injury after mitral valve repair. We report the application and early outcome of this technique for mitral valve regurgitation. METHODS: From January 1997 to January 2004, 16 patients with mitral valve regurgitation were operated on using leaflet folding plasty. The group comprised 9 men and 7 women, with a mean age of 61.6 years. There were 15 patients with degenerative and 1 with ischemic mitral valve disease. The causes of mitral regurgitation were posterior mitral leaflet prolapse in 11 patients and commissural prolapse in 5 patients. RESULTS: Mitral valve reconstruction could be performed in all patients. There were no perioperative deaths. Postoperative mitral regurgitation fell to 0.13+/-0.52 compared with 3.6+/-0.51 preoperatively. Systolic anterior leaflet motion was not observed in any patients after the procedure. The mean follow-up period was 22 months. There were no late deaths and reoperation was not required during follow-up. CONCLUSIONS: Early outcome of leaflet folding plasty for mitral valve repair was satisfactory. This technique may have advantages to accomplish mitral valve repair safely in patients with mitral regurgitation due to posterior or commissural prolapse. PMID- 15782567 TI - Surgical treatment for complications of acute myocardial infarction. AB - OBJECTIVE: We report the results of surgical repair for postinfarction left ventricular free wall rupture (LVFR) and ventricular septal perforation (VSP) complicating acute myocardial infarction. METHODS: We experienced 14 LVFRs and 10 VSPs from January 1991 to December 2002. The mean age of patients with LVFR was 74+/-8.1 years. There were 6 oozing cardiac ruptures and 8 blowout cardiac ruptures. An intraaortic balloon pump (IABP) was inserted in 8 patients, and percutaneous cardiopulmonary support (PCPS) was inserted in 5 patients. The mean age of patients with VSP was 72+/-5.1 years. The rupture was located within the anterior septum in 8 patients and within the posterior septum in 2 patients. All patients required IABP, and one of them needed PCPS. We employed a sutureless technique in 8 patients, direct closure in 5 patients, and infarct exclusion in 1 patient with LVFR. Infarct exclusion was conducted in 9 patients, and the da Silva technique was used in 1 patient with VSP. RESULTS: The patient survival rate of LVFR was 36% (blowout 13%; oozing 67%). Residual shunt occurred in 4 patients with VSP postoperatively. The overall survival rate for VSP was 70%. CONCLUSION: The operative prognosis of blowout cardiac rupture was poor. Good results were obtained with the infarct exclusion technique for patients with VSP. Although some patients had postoperative residual shunts, the infarct exclusion technique was generally a safe and excellent procedure. PMID- 15782568 TI - Impact of multi-detector row computed tomography on the tactics of cardiovascular surgery: from qualitative evaluation to quantitative assessment. AB - OBJECTIVE: We assessed the role of multi-detector row computed tomography in cardiovascular surgery. METHODS: The efficacy of multi-detector row computed tomography was assessed concerning the graft patency of coronary artery bypass, arterial atheromatous degeneration, small vessel imaging, and left ventricular volume measurement. Images were reconstructed using both the volume-rendering and the maximum-intensity-profile methods. Arterial atherosclerotic degeneration was assessed by aortic wall volume and aortic calcification volume. RESULTS: In the assessment of bypass graft patency, multi-detector row computed tomography showed a 98% correct positive ratio with sensitivity and specificity of 98% and 100%, respectively. Atheromatous degeneration showed matching results in more than 70% of cases compared with intraoperative findings. More than 92% of arterial branches with diameters of 3 mm or greater were detected by preoperative multi detector row computed tomography images, though only 6% of branches with diameters of 2 mm or less could be visualized. There was a positive linear correlation between left ventricular volumes determined by multi-detector row computed tomography and those calculated from cine angiography. CONCLUSION: Multi detector row computed tomography clearly visualized coronary bypass grafts and aortic arterial branches, providing detailed vascular images. Atheromatous degeneration assessed by multi-detector row computed tomography was equivalent with intraoperative findings in more than 70% of cases. Left ventricular volumes measured by multi-detector row computed tomography correlated closely with those determined by cine-angiography. Multi-detector row computed tomography is an efficient and promising modality in cardiovascular surgery. PMID- 15782569 TI - Surgical treatment of thoracic aortic aneurysm in patients with concomitant coronary artery disease. AB - OBJECTIVE: Surgical treatment of thoracic aortic surgery in patients with coronary artery disease was investigated. METHODS: Between 1990 and April 2003, 330 patients underwent elective thoracic aortic surgery. Fifty-six patients who underwent aortic root reconstruction were excluded and 274 patients were examined. Fifty-four (20%) patients showed concomitant coronary artery disease. Ten had undergone coronary revascularization previously; and 3 underwent coronary revascularization [2 coronary artery bypass grafting (CABG), 1 percutaneous transluminal coronary angioplasty (PTCA)] before aortic surgery. Twenty-three patients underwent elective CABG simultaneously and 2 patients had additional coronary artery bypass because of cardiac ischemia during operation. The number of patients who underwent thoracic aortic surgery including Asc Ao+AVR was 2, hemi arch 1, total arch 15, distal arch 5, distal arch+LV aneurysmectomy 1, and thoracoabdominal Ao 1. Two patients underwent coronary revascularization with arterial grafts and the others with SVG grafts. RESULTS: There was one hospital death (4%). In patients without coronary bypass, 2 patients suffered cardiac ischemic events. CONCLUSION: Our thoracic aortic operations with concomitant CABG using SVG were overall successful. Our current strategies for thoracic aortic surgery in patients with concomitant coronary artery disease include conducting a dipyridamole myocardial perfusion-imaging test first in patients not at risk of coronary artery disease, and if the test is positive, coronary angiography is performed and aggressive coronary revascularization is conducted where possible. PMID- 15782570 TI - Pseudoaneurysm of the brachiocephalic artery. AB - Pseudoaneurym of the brachiocephalic artery is uncommon. A 62-year-old male patient with a mediastinal tumor underwent tumor resection at the Department of Thoracic Surgery in our hospital. Urgent transverse aorta-right common carotid artery bypass with an 8-mm prosthetic graft was performed intraoperatively, due to uncontrolled bleeding from the brachiocephalic artery where severe adhesion between the anterolateral aspect of the brachiocephalic artery and the tumor was encountered. After surgical removal of the tumor, frozen-section analysis revealed no malignant cells in the tumor, which instead comprised hematoma with organized changes and no evidence of malignancy. Based on pathological results, the resected tumor was diagnosed as a pseudoaneurysm of the brachiocephalic artery. Postoperative course was uneventful. Cause of the pseudoaneurysm was considered iatrogenic, as the aneurysmal wall displayed no atherosclerotic degeneration, and although no history of blunt trauma to the chest or central venous puncture was present, the patient had undergone cerebral artery angiography via the right brachial artery following right thalamic hemorrhage 2 years earlier. PMID- 15782571 TI - Successful treatment of subarachnoid-pleural fistula using pericardial fat pad and fibrin glue after chest wall resection for lung cancer. AB - A 46-year-old male underwent a right adrenalectomy and a left upper lobectomy with an en-block resection of the involved chest wall. The proximal rib resections (third and fourth ribs) were performed at the costvertebral joints. Hemorrhage occurred from near the 3rd intervertebral foramen. The bleeding site was packed with oxycellulose to control the bleeding. Two hours after the operation, the patient complained of paraplegia due to spinal cord compression caused by swollen oxycellulose. An emergency operation was performed. The oxycellulose was carefully removed from the intervertebral foramen. Cerebrospinal fluid was exudated from the spinal canal, however, suggesting an iatrogenic subarachnoid-pleural fistula (ISPF). Autologous fat fragments individually combined with fibrin glue, was packed gently into the intervertebral foramen. The orifice of the foramen was then covered with a pericardial fat pad. The postoperative course was uneventful. This new technique is a direct and effective treatment for an ISPF. PMID- 15782572 TI - Primitive neuroectodermal tumor/Ewing's sarcoma with long-term follow-up. AB - We have reported a case of primitive neuroectodermal tumor/Ewing's sarcoma (PNET/EWS) in a 32-year-old female who had had no symptoms until the tumor was detected in a general examination after she gave birth. Chest X-ray showed a well defined, rounded mass in the left lower lung field. Computed tomographic scan of the chest revealed a distinct mass adjacent to the anterolateral thoracic wall. The macroscopic findings had shown a well-capsulated tumor with a stalk emanating from the thoracic wall. PNET/EWS was histologically diagnosed. The tumor was composed of atypical, small round cells. It stained positively with antibodies to the MIC2 gene product. Postoperative adjuvant radiotherapy of 50 Gy was performed. Adjuvant chemotherapies were not performed. Four years later, a small recurrent nodule in the left lung was surgically resected. Since her initial diagnosis, the patient has survived for seven years; in the three years since her second surgery, she has had no other recurrence or metastasis. PMID- 15782573 TI - Primary lung adenocarcinoma with heterotopic bone formation. AB - A 46-year-old man was referred to our hospital for the treatment of lung cancer. Computed tomography showed a well-defined tumor mass that was 50x45 mm in size and contained a trabecular pattern of calcification. Since he was diagnosed as having a primary lung adenocarcinoma (clinical stage IB), a left upper lobectomy with mediastinal lymph node dissection was performed. Histologically, the tumor was a poorly differentiated adenocarcinoma with rich fibrous stroma, in which there were island-shaped bone formation lesions. An immunohistochemical examination showed the expression of bone morphogenic protein-2 within tumor cells, which induce and stimulate bone formation. This finding may elucidate a possible mechanism of heterotopic bone formation. PMID- 15782574 TI - Primary chondrosarcoma of the lung recognized as a long-standing solitary nodule prior to resection. AB - As the use of computed tomography (CT) increases, incidental lung nodules have become a clinical issue that is being addressed more than before. We detected a solitary lung nodule which was smooth-margined, round-shaped, 11 mm in size. Follow-up for 18 months after initial detection by chest CT did not show any interval change. To make a definitive diagnosis, video-assisted thoracic surgery was performed and the lesion was diagnosed as myxoid chondrosarcoma. In the 6 year postoperative follow-up, annual chest CT and bone scintigram did not reveal any abnormality, which excludes the possibility of a latent primary site other than the lung. Therefore, we considered the present case being of pulmonary origin. Accordingly, even though the lesion appeared unremarkable, surgical resection of solitary lung nodule should not be discouraged. PMID- 15782575 TI - Coronary artery and graft spasm after off-pump coronary artery bypass grafting. AB - We report a case of a 52-year-old man with severe coronary artery and graft spasm after triple-vessel off-pump coronary artery bypass grafting. Emergent coronary angiography was performed to identify the location and severity of the spasm. Intracoronary injections of several vasodilators failed to relieve the spasm. Observational treatments including intra-aortic balloon pump and inotropic drugs to increase coronary flow were performed until the spasm resolved. The patient recovered and was discharged. A follow-up coronary angiography revealed patent native coronary artery and bypass grafts without evidence of residual spasm. PMID- 15782576 TI - Ross procedure for congenital aortic insufficiency and an associated right intramural coronary artery. AB - A 7-year-old boy diagnosed with aortic insufficiency was treated with a Ross procedure and he had an associated right intramural coronary artery. Although preoperative angiocardiography showed that the right and left coronary artery orifices were close together, a right intramural coronary artery could not be diagnosed by transthoracic echocardiography. Intraoperative findings showed that the right coronary artery was intramural for a distance of 7 mm. As a result, a longer single coronary button was harvested and transplanted to the pulmonary autograft. The postoperative course was uneventful; coronary ischemia did not occur and aortic valve function was preserved. PMID- 15782577 TI - Dissection and rupture of the left subclavian artery presenting as hemothorax in a patient with von Recklinghausen's disease. AB - Hemothorax is a rare and life-threatening complication of von Recklinghausen's disease. We present a case of a 48-year-old man with this disease who developed hemothorax caused by rupture of a left subclavian artery dissection. To our knowledge, the case presented here is the first to describe successful surgical management--graft replacement of the left subclavian artery--in hemothorax due to arterial dissection associated with von Recklinghausen's disease. PMID- 15782578 TI - Giant left ventricular pseudo-false aneurysm following myocardial infarction. AB - We report a successful surgical case of giant left ventricular pseudo-false aneurysm in a 63-year-old man. The abnormality of the inferior wall of the left ventricle was discovered incidentally by abdominal ultrasonography for health examination at another hospital. Transthoracic echocardiography and left ventriculography revealed a giant false aneurysm (74x75x40 mm) in the inferior wall of the left ventricle with a large orifice (70x58 mm). Repair of the aneurysm was performed under arrested heart, closing the large orifice with a Hemashield patch. Postoperative pathological examination proved that the wall of the aneurysm had myocardial tissue. The microscopic results strongly suggested that this aneurysm was a pseudo-false aneurysm. PMID- 15782579 TI - [Do the Japanese drink less alcohol than other peoples?: the finding from INTERMAP]. AB - It has been pointed out that Japanese men drink more than American and British men. In this presentation, we report the comparison of drinking habits from the findings of the International Study of Macro-nutrients and Blood Pressure (INTERMAP) conducted during 1997-99 in Japan, China, the United States of America (U.S.), and the United Kingdom (U.K.). INTERMAP was conducted with highly standardized procedures including dietary assessment, blood pressure measurement and questionnaires to clarify and compare their relationship in randomly selected 4680 men and women aged 40-59 years from 17 centers in the four countries. For alcohol drinking habits, four times of 24-hour dietary recall studies and two times of one-week recall studies were conducted. The drinking prevalence of Japanese men was the highest among the four countries. For women, it was also higher than in U.S. but lower than in U.K. Abstinence rates in U.S. were extremely higher than those of Japanese (19, 20% vs. 1, 4% in men and women, U.S. vs. Japan, respectively). Japanese men consumed the average of 27 g per day of alcohol and it was the highest among the populations. Energy intake from alcohol per total energy intake was also the highest in Japanese men. Health professionals in Japan need to recognize from the findings of INTERMAP and other studies such as INTERSALT that Japanese men drink more alcohol than Americans and British counterparts. PMID- 15782581 TI - [Alcohol dependence syndrome and BDIM (before-discharge intervention method)- report 4, the family members' self-reports about BDIM]. AB - One hundred fifty-three inpatients with alcohol dependence syndrome were treated with the structured BDIM (Before-Discharge Intervention Method). 82 patients of them have participated to self-help group meetings or kept having therapy as our outpatients or inpatients during the study period. We chose the families of the 82 patients as our study subject Out of the study subjects who took part in BDIM, 64 families (117 persons) answered our questionnaire. Among them 63 families (101 persons) gave their described answers of impressions and opinions about BDIM, which were summarized as follows. (1) Through BDIM the family members gained second thought on their alcoholic family member (IP: identified patient) and they could tell their new view to IP. BDIM enabled them to tell IP their sincere feeling and hope for recovery of IP. BDIM empowered both IP and IP's family members. (2) The family members became to know IP's orientation on his or her disease. They came to know IP's denial and understand him or her as he or she was. (3) The family members felt emotional ties among themselves and IP through BDIM. When the family members of a dysfunctional family took part together in BDIM, they could know the feelings, thoughts, experiences and hopes one another. The family members had a precious experience of mutual understanding among themselves and IP to hope for recovery together. (4) The family members appreciated BDIM as a effective therapy. In BDIM many of them regarded highly of giving their letters to IP as a useful method to convey their feeling and thoughts calmly to IP. (5) On the other hand some family members pointed out the difficulty for themselves to write on BDIM. For family members who are not good at writing a letter or tend only to blame IP through their letters, writing and giving letters to IP is not appropriate as a therapy. If family members feel strong anxiety or fear, it is safe not to practice BDIM. PMID- 15782580 TI - [Study on the perception of influencing factors in the course of alcoholism--a comparison among alcoholics, families, and health care professionals]. AB - (1) We prepared a questionnaire modified from that of Edwards, G. et al, to investigate the factors influencing drinking behaviors. Responses of the questionnaire were filled out by the alcoholics, their families, and health care professionals in four psychiatric hospitals having intensive alcoholic rehabilitation programmes. Clinical course of them were traced back for at least two years. (2) We examined the relationship between the course of alcoholism and the differences in perceptions among alcoholics, their families, and health care professionals. (3) We obtained sufficient responses from 76 alcoholics. Those were divided into two groups; one is a group of alcoholics with a history of continuous abstinence of over one year (50 cases), and the other is the group of less than one year of recent abstinence period (26 cases). (4) As the overall results from the participants, items on "self-help group" were favorably responded for the improvement of drinking behavior. Families tend to perceive more sensitively into the internal psychological changes of the alcoholics than the health care professionals, whereas the health care professionals tend to make general and common responses regardless of any alcoholics. We, health care professionals should be aware of such a limit to promote more efficient collaborative works with alcoholics and their families. (5) We compared these two groups ("over one year group" vs. "less than one year group") with regard to their demographic and family backgrounds, clinical features, treatment, self-help group attendance, employment, etc. (6) A significant accordance of responses among three groups (alcoholics, family, health care professionals) was found in the "over one year group" than in the "less than one year group". Such a high level of accordance is considered to reflect a favorable family adaptation along with the recovery from alcoholism, deep understanding on the part of health care professionals, progress in the treatment and in the relationship in a self-help group. (7) As regards the "less than one year" group, it includes more single individuals and less frequent attendances to self-help groups. For this group of alcoholics, group therapy in a smaller group, individualistically oriented approaches for their needs including various supportive resources in the society should be addressed. PMID- 15782582 TI - Case of prolonged alcohol withdrawal syndrome accompanied with hyperthyroidsim. AB - Hyperthyroidism is associated with increased psychiatric morbidity. It may alter the clinical course of alcohol withdrawal syndrome. We report a 69 year old man who presented prolonged alcohol withdrawal syndrome associated with hyperthyroidism. Initially, he developed typical alcohol withdrawal syndrome including tremor, disorientation, delirium and visual hallucination of small animals. Thyroid function tests revealed a free triiodothyronine (T3) of 6.1 pg/dl (range, 3.0 to 5.8), a free thyroxine (T4) of 2.3 ng/dl (range, 0.85 to 2.15) and a thyroid stimulating hormone (TSH) of 0.003 microU/ml (range, 0.3 to 4.0), and thiamazole was administered. Even after a month, he continuously presented persecutory delusion, auditory hallucination and cognitive dysfunction. Although these symptoms did not respond to the medication including antipsychotics, they totally passed away after the thyroid function reached down to the normal level (free T3 3.0 pg/ml, free T4 1.1 ng/dl, TSH 0.004 microU/ml). In addition, cognitive function was recovered to the normal level as he scored 28/30 on the Mini Mental State Examination. We propose that hyperthyroidism contributed to the occurrence of psychotic symptoms and cognitive dysfunction. PMID- 15782583 TI - [Effects of specificity of schema on false recall: an analysis from the viewpoint of eyewitness testimony]. AB - The purpose of this study was to investigate the effects of two levels of specificity of schema on generation of false recall. One level is widely applied to things and is shared by the general public (less specific schema) and the other is specifically applied to individual things (more specific schema). Sixty female undergraduates watched a video. After two days, they were required to recall the contents of the story. Students were divided into two groups according to whether they have the more specific schema or not. In each group, they were assigned to either free recall task or reality monitoring task. The results showed that (1) the amounts of false recall by the group having the less specific schema only decreased by reality monitoring. (2) The group having the more specific schema had no differential effects on false recall for both tasks. (3) The effect of specificity of schema on false recall was not observed for the scene which did not activate the more specific schema. These results were discussed in terms of the levels of specificity of schema and effectiveness of reality monitoring for eyewitness memory. PMID- 15782584 TI - [Roles of reader's feelings in understanding narratives: forefeel, empathy, and a sense of strangeness]. AB - This study examined the cross-domain, anticipatory, and self-referential roles of affect (Miall, 1989) in reading short mystery novels. In Experiment 1, undergraduate students either read an entire story once or read the same story twice. During the first reading, these participants read only the first part of the story, and during the second reading, they read the entire story. They rated the importance of each sentence. Then they either described a prediction or their impression of the story. Finally, they created a title. In Experiment 2, the first group rated the importance of each sentence. The second group rated the feelings of understanding: forefeel, empathy, and a sense of strangeness. The results of both experiments showed that the relative importance of sentences shifted between first reading and second reading. As they read the end of the story, empathy increased but a sense of strangeness decreased. The results of description showed that the readers updated their interpretations in order to understand the story correctly. PMID- 15782585 TI - [Effects of the diversity and relative dimension of the peripheral objects on the meaning of the qualifying word for young children]. AB - Two experiments were conducted to test 100 children (aged five years) on an object named novel word selection task. In Experiment 1, the experimenter labeled a novel word to a target object (an elephant) which was in a cage alone, or which was in a cage with many objects (other animals and instruments) around it. In the former case, the subjects usually qualified the word meaning only when similarly restricted by its peripheral objects (a cage). In the latter case, the subjects were likely to qualify the word meaning with a conceptual level name (an elephant). In Experiment 2, the experimenter labeled a novel word to a target object which was in a small cage, or in a large cage. In the former case, the subjects were likely to qualify the word meaning only when being similarly restricted by its peripheral objects. In the latter case, the subjects were likely to qualify the word meaning with a conceptual level name. These findings suggested that, for young children, objects peripheral to a target object had an effect on the meaning of the qualifying word. PMID- 15782586 TI - [Study of the effect of interpersonal stress coping training on reducing stress response: a research of student nurses]. AB - This study investigated, the effect of interpersonal stress coping training on reducing stress response of 100 student nurses. They were divided into two groups; the coping training (CT) group and the no training control (NT) group. During a three-week period, 50 participants in the CT group were required to adopt postponed-solution coping and keep daily records of how they coped with interpersonal distress. All participants were asked to complete a self-reported questionnaire including measures of distress and coping strategies before and after the training, and also after a 18-week follow-up period. The CT group, compared with the NT group, significantly improved their postponed-solution coping after the training. The CT group was found to be significantly lower in distress than the NT group both after the training and after the follow-up period. PMID- 15782587 TI - [Environmental context effects of background colors on recognition memory]. AB - Three experiments examined whether or not switching study background-color contexts among target words at testing reduces word-recognition performance. These experiments also examined whether or not presentation rate--one of the determinants of item strength--interacted with background-color context. Undergraduates learned 40 target words presented at a rate of 1.5 or 3.0 seconds per word in one of two background-color contexts in Experiment 1, and in one of ten contexts in Experiments 2 and 3. Recognition of the targets was tested by mixing 40 distractor words with the targets immediately after the learning session in Experiments 1 and 2, and with a 5-minute filled retention interval in Experiment 3. Experiment 1 failed to find background-color context effects on recognition, but Experiments 2 and 3 successfully found the context effects. Presentation rate did not interact with the context effects. The results conflict with the ICE theory. The implications of the present findings are discussed. PMID- 15782588 TI - [The differences in young and elderly adults on false memory: examination on the effect of retention interval]. AB - False recognition can occur at high levels after participants study lists of associated words and are tested with semantically related lures. The present study attempted to determine the differences of young and elderly adults in retention interval on false recognition and remember-know judgments of critical lure. Young participants' mean age was 21.6 years (range 21-23 years). Elderly participants' mean age was 69.7 years (range 65-74 years). Participants engaged in estimating the frequency of usage of visually presented words, and they were tested both immediately and one week later after the study phase. We found that elderly adults were relatively more susceptible than young adults to this false recognition effect. Moreover, a reliable increase of false recognition for critical lures was observed across retention interval in young adults but not in elderly adults. PMID- 15782589 TI - [Big five personality factors related to face recognition]. AB - The present study examined whether scores on big five personality factors correlated with face-recognition response time in visual search paradigm. Sixty adjectives were used to measure personality scores of 60 participants along the five factors of Extroversion, Neuroticism, Openness to Experience, Agreeableness, and Conscientiousness. Picture of human faces or geometrical figures in a 4 x 4 array were used as stimuli. The sixteen faces or figures were either identical (absent condition) or one randomly placed target with 15 identical distracters (present condition). Participants were asked to respond 'present' or 'absent' as fast and accurately as possible. Results showed that the response time differed significantly between high and low groups of each personality factor except Agreeableness. For Extroversion, Neuroticism, and Conscientiousness, the response time difference was observed only for human face recognition. The results suggested that personality differences and face recognition were related. PMID- 15782590 TI - [An attempt to integrate the dual route cascaded model and the triangle model for reading English words aloud]. AB - In this article, I discuss the implementation of neural network models for reading English words aloud. Since 1989, there has been existing a debate about the models of reading English words aloud. One is the Dual Route Cascaded (DRC) model. The other is the Triangle model, whose original version was developed in 1989. Because there are arbitrary variables of both models, we did not decide which model gives better accounts for the numerous data given by psychological experiments and neuropsychological evidence. Therefore, in order to give a solution of this debate, an attempt to integrate both models was made. Introducing the Mixture of Experts Network, an elegant solution to overcome the arbitrariness of both models could be given. The Mixture of Expert Network can include both the models as a special case of this Network. From the Mixture of Expert Network's point of view, the difference between the Dual Route Cascaded model and the Triangle model would be able to describe as the quantitative difference of the dispersion parameters. PMID- 15782591 TI - [Study on self and other ratings of hostility]. AB - The purpose of this study was to investigate the validity of hostility ratings made by others. The rating measure was considered essential for the study of correlation between hostility and coronary heart disease. Undergraduate and graduate students, 81 in all, completed a questionnaire of hostility and social desirability. The scales included were Buss-Perry Aggression Questionnaire (BAQ), Muller Anger Coping Questionnaire (MAQ), and Marlowe-Crowne Social Desirability Scale (MCSD). Acquaintances also rated them using BAQ and MAQ. Correlations between self and other ratings were moderate, and self-rated hostility scores correlated negatively with MCSD scores, while other-rated hostility scores did not. Results of ANOVA indicated that self-rated hostility scores were higher than other-rated hostility scores. However, the differences decreased as the degree of closeness between them increased, and no difference was found in long-term close relationship. These results suggested that other ratings of hostility could be an objective measure in Japan, as in western countries. PMID- 15782592 TI - [Relationship between members' egoistic self-assertion and captains' leadership in athletic clubs]. AB - This study examined the relationships between club members' egoistic self assertion and their captains' leadership with respect to the adjustment to activities in extracurricular athletic clubs. Participants were 638 junior high school students who belong to the athletic clubs. The research was conducted by means of a questionnaire and the following results were obtained. When the captains took a laissez-faire command, club members who did not assert themselves egoistically were not satisfied with their captains, and did not participate in the club activities very diligently. On the other hand, when the captains took an authoritative command, the members who asserted themselves egoistically were not satisfied with club activities. These results suggest that the relationships between egoistic self-assertion by club members and leadership by captains have a great influence on the adjustment to club activities. PMID- 15782593 TI - [Effect of orthography on the usage of recognition heuristic]. AB - This study examined the effect of orthography (kanji, kana, and alphabet) on the usage of recognition heuristic, in the context of ecological rationality research. Forty-five students participated and performed the task consisted of randomly drawn two-alternative questions about population sizes of the 20 largest Chinese cities, except Beijing and Shanghai. It was found that kanji and kana tasks led to the usage of recognition heuristic more than alphabet task. Furthermore, judgments in kanji and kana tasks were more accurate than in alphabet task. These results suggested that the usage of recognition heuristic was facilitated when it increased judgment accuracy, and that recognition heuristic had ecological rationality. PMID- 15782595 TI - [Therapeutic cerebral hypothermia after Clifton shock in 2001]. PMID- 15782596 TI - [Can diffusion tensor imaging predict the functional outcome of supra-tentorial stroke?]. AB - We used diffusion tensor imaging (DTI) to assess wallerian degeneration of the pyramidal tract after the onset of supra-tentorial stroke, and correlation of the extent of Wallerian degeneration with the motor function at 3 months after stroke. Twenty eight patients with supra-tentorial acute stroke were examined, two weeks and one month after stroke by DTI. We measured fractional anisotropy(FA) of affected side/ unaffected side (FA ratio) in the cerebral peduncle. We used modified Rankin Scale (mRS) for assessment of motor function at 3 months after stroke. FA ratio was significantly reduced at 2 weeks after stroke (0.833 +/- 0.146) compared to on admission (0.979 +/- 0.0797). But no significant change of FA ratio was seen between two weeks and one month after stroke in 7 cases examined (0.758 +/- 0.183 vs. 0.754 +/- 0.183). In all patients in whom the FA ratio was under 0.8 at 2 weeks after stroke, motor function showed poor recovery (mRS 4 and 5) at 3 months after stroke. When FA ratio was over 0.8 at 2 weeks after stroke, motor function at 3 months after stroke showed good recovery (mRS 0 to 3) expect for three elderly patients. With the use of DTI, Wallerian degeneration could be detected in the corticospinal tracts at midbrain level during the early phase of supra-tentorial stroke. We conclude that DTI may be useful for early prediction of motor function prognosis in patients with supra tentorial acute stroke. PMID- 15782594 TI - [Channelopathy update]. PMID- 15782597 TI - [Vasospastic cerebral infarction induced by smoking: a case report]. AB - A 31-year-old man was admitted to our hospital because of frequent transient ischemic attacks (TIAs). The first episode involved right amaurosis fugax and left hemiparesis at the age of 26. Treatment with aspirin did not reduce frequency of TIA. Cerebral angiography at the age of 29 showed a significant stenosis in the right internal carotid artery with a string-of-beads-like appearance. This pattern suggested fibromuscular dysplasia. TIAs persisted despite of prophylactic medication with ticlopidine. When cerebral angiography was repeated at age of 28, stenosis in the right internal carotid artery had almost disappeared. At the present admission, MR angiography showed stenoses of bilateral internal carotid arteries and middle cerebral arteries, which had disappeared when the study was repeated after 5 days. Vasospasm was suspected based on reversibility of changes in both conventional and MR angiographies. The patient was treated with a calcium antagonist to prevent vasospasm as well as cessations of smoking. The patient had a history of 20 cigarettes a day for 12 years and neurologic deficits often occurred after smoking. Therefore, smoking is considered to be a main trigger for TIAs in this patient. PMID- 15782598 TI - [Medulloblastoma with mandibular metastasis after complete remission of the central nervous system lesions]. AB - A 6-year-old boy presented with headache and vomiting. Brain and spinal MRI demonstrated a large mass in the cerebellar vermis and 4th ventricle and showed thick spinal subarachnoid dissemination. Suboccipital craniotomy was performed and the tumor was totally removed. The histological diagnosis was medulloblastoma. The patient subsequently received craniospinal irradiation, and also received systemic and intrathecal perfusion chemotherapy. Then complete remission was achieved 10 months after operation. Three years later, however, a swelling at the left mandibular angle appeared. A CT scan revealed osteosclerotic lesion. After biopsy was performed, the specimen was detected infiltration of medulloblastoma cells. Bone scintigram showed a single lesion and MRI of brain and spinal cord revealed no recurrence of the central nervous system lesions. He underwent local irradiation and systemic chemotherapy with ICE regimen. This is the rare case of extracranial metastasis following remission of medulloblastoma in childhood. PMID- 15782599 TI - [Jargonagraphia in a left-handed aphasia due to a left hemisphere lesion]. AB - We report a case who developed jargonagraphia, severe aphasia and unilateral spatial neglect due to cerebral infarction in the left hemisphere. The patient was a 57 year-old left-handed woman. She suddenly developed hemiplegia on the right side and aphasia, and was admitted to our hospital. Neuropsychological examination showed non-fluent spontaneous speech. Repetition, reading and auditory comprehension were impaired. Writing was severely impaired and she showed paragraphia without meaning (jargonagraphia). Right unilateral spatial neglect and buccofacial apraxia were noted, but neither ideomotor nor ideational apraxia was observed. The mechanisms of jargonagraphia remain unknown. The localization of language function in this patient is not a mirror image of same function in dextral aphasia. Her condition was considered that free-running on motor engrams of characters stored in the right hemisphere caused jargonagraphia. PMID- 15782600 TI - [A case of catatonia associated with the ingestion of hydroxyzine]. AB - A twenty-six-year male who presented with a consciousness disturbance induced by the ingestion of an estimated 7g of hydroxyzine. He demonstrated bipolar symptoms which consisted of both stupor and excitement, and both an increase in muscle tension and apnea, however, these symptoms improved after the infusion of diazepam. He was diagnosed to demonstrate catatonia. After treatment composing three days of mechanical ventilation in combination with the administration of sedatives and muscle relaxatants, his symptoms improved. Hydroxyzine is thus considered to be able to induce catatonia and this mechanism of this condition is discussed. PMID- 15782601 TI - [Syndrome of inappropriate secretion of antidiuretic hormone (SIADH) associated with relapsing multiple sclerosis]. AB - We report a 47-year-old man with multiple sclerosis (MS) with previous history of recurrent sensorimotor disturbance and visual deficit. The patient developed bilateral motor weakness in the upper limbs, and systemic malaise. An administration of 20 mg/day of prednisolone was ineffective for his symptoms and he complained dyspnea a week later. On admission, his clinical findings included brainstem dysfunction with optic nerve atrophy, motor disturbance in the bilateral upper limbs, hyperreflexia, and superficial sensory disturbance. Biochemical examination revealed marked reduction in serum Na (117 mEq/l) and C1 (85 mEq/l) with increased urinary Na excretion. Although his plasma osmotic pressure decreased to 233 mOsm/kg, urinary osmotic pressure increased to 409 mOsm/kg. Serum antidiuretic hormone (ADH) concentration was 26.1 pg/ml and plasma renin activity was 0.1 ng/ml/ hour. Renal function and adrenal function were normal. Cerebrospinal fluid contained increased protein concentration, IgG, and myelin basic protein. Syndrome of inappropriate secretion of antidiuretic hormone (SIADH) associated with MS was diagnosed. Intravenous Na infusion with restricted supplemental fluid and serial administration of methylprednisolone (1,000 mg/day for three days) improved his neurological abnormalities and normalized his serum serum Na level and plasma osmotic pressure. This suggests that demyelinating lesions in the hypothalamus due to MS may cause the transient increased ADH secretion. PMID- 15782602 TI - [Prolonged cerebral salt wasting following craniopharyngioma surgery and posterior reversible encephalopathy syndrome: a case report]. AB - A 9-year-old boy was admitted to our hospital with daytime urinary incontinence for the past one year. MRI showed craniopharyngioma occupying the third ventricle. The tumor was excised by interhemispheric approach. Because hyponatremia and polyuria with high renal loss of sodium were observed on postoperative day 3, hydrocortisone and DDAVP were replaced. On postoperative day 24, successive general convulsions and hyponatremia recurred, and MRI FLAIR imaging showed marked brain edema in the bilateral parieto-occipital lobes. This finding disappeared late in the course of treatment, and the case was diagnosed as posterior reversible encephalopathy syndrome. The pathophysiology of cerebral salt wasting and posterior reversible encephalopathy syndrome in a craniopharyngioma patient are also discussed in the article. PMID- 15782603 TI - [Sumatriptan nasal spray for the treatment of cluster headache]. AB - We report three cases with cluster headache treated with sumatriptan nasal spray. In all occasion, sumatriptan nasal spray showed meaningful relief in patients with cluster headache, although the efficacy were less than that of sumatriptan subcutaneous injection. Sumatriptan nasal spray needed 20 to 30 minutes to meaningful relief, while subcutaneous injection needed only 5 to 10 minutes. Recurrence of attack was more frequent in oxygen inhalation therapy than sumatriptan nasal spray. Sumatriptan nasal spray were constantly effective in both early stage and late stage of cluster periods. Sumatriptan nasal spray may shorten the admission period of severe case of cluster headache. Sumatriptan nasal spray may enable doctors to treat mild cases of cluster headache in their outpatient clinic. PMID- 15782604 TI - [A 65-year-old man with rigid-bradykinetic parkinsonism, vertical gaze palsy, difficulty of eye-lid opening, and marked pseudo-bulbar palsy]. AB - We report a 65-year-old man with rigid-bradykinetic parkinsonism, vertical gaze palsy, difficulty in eye-lid opening, and marked pseudo-bulbar palsy. He felt difficulty of it, hand movement at 59 years old. When he was 60 years old, monotonous speech and slowness of movement appeared. He visited a neurologist who noted vertical gaze palsy, neck rigidity, and bradykinesia. He was diagnosed as progressive supranuclear palsy (PSP) and given 300 mg L-Dopa/Benserazide by the neurologist. This medication improved his rigidity and bradykinesia. At 62 years of the age, his eye-lids closed involuntary and it was difficult to open. In addition, he began to complain of wearing-off, autonomic symptoms, and dysphagia. Anti-parkinsonian drugs were increased, but his bradykinesia progressed. At 64 years of the age, he was admitted to the Neurology Service of Juntendo Hospital. On admission, he was alert and not demented. No aphasia, apraxia, or agnosia was noted. In the cranial nerves, upward and downward gaze were markedly restricted. His face was hypomimic and seborrhoic. It was difficult to swallow liquid or solid for him. No weakness was noted, but he walked in small steps with freezing and falling tendency to backward. Rigidity was noted on his extremities and stronger on his left side than right. Tremor was absent. Bradykinesia of his body and extremities was marked. No cerebellar ataxia was noted. Deep tendon reflexes were within normal range. Planter response was flexor bilaterally. Myerson's sign was noted. Sensory and autonomic function were normal. He was treated with L Dopa, Pergolide, and Bromocriptine. However, these medications improved his bradykinesia and gait disturbance only slightly, dysphagia became progressively worse. He developed aspiration pneumonia when he was 65 years old and admitted to Juntendo Hospital. A large amount of sputum was aspirated from his trachea. Two days after from admission, he was found dead on his bed. He was discussed in a neurological CPC and the chief discussant arrived at a conclusion that the patient had progressive supranuclear palsy (PSP). Other differential diagnoses included Parkinson's disease, pallido-nigroluysian atrophy (PNLA), multiple system atrophy (MSA), and corticobasal degeneration(CBD). Many participants considered that PSP or PNLA was most likely. Post-mortem exmination revealed marked nigral neuronal loss and gliosis. The globus pallidus and the luysian body changed mildly. However, the frontal cortex was relatively spared, there were many ballooned neurons in the cortical layer. Other parts were spared. With sliver (Bodian and Gallyas-Braak) and anti-phsphorylated tau stain, abundant astrocytic plaques, neurofibrillary tangles, and argyrophilic threads on the frontal cortex, striatum, and substantia nigra were seen. There was no tufted astrocyte which was hallmark of diagnosis of PSP. In addition, several Lewy bodies were seen in the brainstem. Because astrocyte plaque was considered specific for pathology of CBD, the pathologist revealed that the pathological diagnosis of this patient was CBD. Nevertheless, discussion was focused on the relatively mild degeneration of the frontal cortex for CBD. PMID- 15782605 TI - [Clinical investigation of the 8 cases with AIDS (acquired immunodeficiency syndrome)-associated progressive multifocal leukoencephalopathy (PML)]. AB - We experienced 8 cases of progressive multifocal leukoencephalopathy (PML) complicated by human immunodeficiency virus (HIV) type-1 infection from 1985 to 1999. These cases showed dementia, bradykinesia, dysarthria, hemiparesis, and so on. All of the cases were severely immunocompromised hosts, because none had more than 150/mm3 CD4 + lymphocytes; indeed, 5 of the cases were below 20/mm3. Other neurological complications except PML were primary CNS lymphoma, HIV encephalitis, and CMV encephalitis. The mean life durations was 7.6 months after the first symptom appeared, for all but one of the patients; the exceptional patient lived for 24 months after. Autopsy studies of the central nervous systems were performed for 7 cases, all of which showed extensive demyelinating lesions of the white matter, and in some cases these extended into the spinal cord. In contrast to Western countries, in Japan there have been few reports of AIDS associated PML. Thus, this report was thought to be important here. PMID- 15782606 TI - [Primary progressive multiple sclerosis as a differential diagnosis of ALS: a case report]. AB - A 44-year-old man was admitted to our hospital because of a five year history of chronic progressive gait disturbance. Neurological examination revealed mild weakness and atrophy of the upper extremities, but severe of the lower ones, and without sphincter disturbance or apparent sensory impairment. Hyperreflexia and positive pathological reflexes of the lower extremities were apparent. EMG showed a reinnervation pattern and decreased number of motor units in the extremities, suggesting ALS. However, multiple plaques on the head and spinal MRI, a prolonged central conduction time of MEP and SEP, a delayed P100 latency of VEP, and a increased IgG index in the CSF indicated primary progressive type multiple sclerosis. After receiving steroid pulse therapy, the weakness of the lower extremities showed slight improvement. Diffuse inflammation in the spinal cord involving not only the pyramidal tract but also the anterior horn cells/intramedullary ventral roots explained the ALS-like clinical picture. PMID- 15782607 TI - [An autopsied case of limbic encephalitis associated with extremely high titers of anti-SS-A antibodies in serum and anti-neuronal antibodies in CSF]. AB - A 76-year old man was referred to our department because of several episodes of generalized convulsion followed by a loss of consciousness and the right hemiparesis. The disturbed consciousness and hemiparesis disappeared soon but the personal change persisted thereafter. T2 and diffusion weighted images of MRI taken on the admission showed high intensity lesions in the left medial temporal lobe including the hippocampus. Antibodies (Abs) against herpes simplex virus were not elevated, however, serum titers of antinuclear and anti-SS-A/Ro Abs were extremely elevated. CSF IgG level and IgQ index were increased, and the CSF reacted with 78-kd bands on Western blots of rat brain homogenate. He died of bacterial pneumonia on the 28th day of illness and was autopsied. Malignant tumors were not found in any organs. In the left hippocampus, degeneration and loss of neurons, infiltration of macrophages, and microgliosis were observed. Vasculitis, however, was not found in the lesion. The immunohistochemical study showed that the CSF recognized the cytoplasm of neurons in the human hippocampus and also Purkinje cells. Those immunological and pathological findings thus suggest an antibody-mediated autoimmune limbic encephalitis in our case. PMID- 15782608 TI - [Toxoplasma encephalitis in a patient receiving cyclosporine monotherapy for Behcet disease]. AB - We report a 44-year-old woman with toxoplasma encephalitis that occurred during cyclosporine monotherapy for Behcet disease. She had been treated with cyclosporine for 8 years. She experienced headache and, nausea, and then consciousness disturbance developed. Brain MRI showed high-signal intensity lesions on T1-weighted MRI with Gd-enhancement in the left temporoparietal lobe, right thalamus and right frontal and temporal lobes. The pathological examination of the biopsied brain specimens suggested toxoplasma encephalitis. She improved rapidly after the administration of antibiotics for toxoplasma gondii. Anti toxoplasma specific protein antibodies were positive in the serum and CSF, supporting a diagnosis of acute toxoplasmosis. Toxoplasma encephalitis due to cyclosporine mono-therapy has not been reported yet. The measurement of anti toxoplasma specific protein antibodies may be useful for the early, accurate diagnosis of toxoplasmosis. PMID- 15782610 TI - [Perception of dyspnea due to breath-holding in myotonic dystrophy]. AB - Patients with myotonic dystrophy (DM1) rarely complain of dyspnea despite of severe hypoxemia. We studied the perception of dyspnea caused by breath-holding in 9 DM1 patients and 8 healthy control subjects. The patients, as well as the control subjects, complained of dyspnea and showed decrease in SpO2. In none of the patients but one, however, the bottom SpO2 became lower than the minimal SpO2 recorded in 24-hour monitoring. DM1 patients were able to realize hypoxia caused by apnea, although they had not realized hypoxia that already existed. Consequently, the breath-holding test does not uncover a blunted perception of dyspnea in DM1; an afferent system contributable to air hunger sensation in breath-holding is preserved in DM1. Breath-holding test may be useful for a DM1 patient to recognize the significance of sleep apnea. PMID- 15782609 TI - [Clinical, neuroimaging and electroencephalographic findings of encephalopathy occuring after the ingestion of "sugihiratake" (Pleurocybella porrigens), an autumn mashroom: a report of two cases]. AB - We report a 54-year-old man (case 1) and a 79-year-old woman (case 2) who presented with encephalopathy of unknown cause. Both patients were on hemodyalysis and took an autumn mashroom, "sugihiratake" (Pleurocybella porrigens), two to three weeks prior to the onset of neurological alterations. The clinical syndrome of those patients was characterized by weakness and involuntary movements of the extremities (cases 1 and 2) or dysarthria (case 1) at the onset of the disease and subsequent intractable focal motor seizures, resulting in generalized status epilepticus or comatose state, six (case 1) or three (case 2) days after the disease onset. Epileptic seizures were gradually improved in both cases. On brain MRI of case 1, no relevant lesions were detectable at the onset day, but, 6 days after onset, T2-high intensity lesions were noted in the subcortical white matter of the insular cortex, claustrum, external capsule, putamen and globus pallidus on both sides. On brain CT scan of case 2, there were no apparent lesions at the onset day, but, 4 days after onset, low density areas were noted bilaterally in the subcortical white matter of the insular cortex. Electroencephalography of the two patients taken on a day of comatose state showed periodic synchronous discharge (PSD), which disappeared when their consciousness levels were improved. As far as we have examined, there was no findings to suggest the cause of the encephalopathy in routine laboratory examinations and various viral antibody studies of the blood and cerebrospinal fluid. The reported patients could constitute a newly recognized disease entity, "sugihiratake" encephalopathy. Our observations suggest that it can be an encephalopathy with subacute progression and affect mainly the basal ganglia. Neuroimaging study and electroencephalographic findings may help the diagnosis, although they may remain unremarkable for several days after onset of the neurological alterations. PMID- 15782611 TI - [Effectiveness of tacrolimus in a case of polymyositis]. AB - We report a 57 year-old woman with polymyositis and interstitial pneumonia. With the steroid therapy alone, the decline of creatine kinase was insufficient and muscle strength was not improved. After the addition of tacrolimus 3 mg, serum creatine kinase declined, muscle strength was improved, and the dose of steroid could be tapered very smoothly. There was no remarkable change in the status of the interstitial pneumonia on the chest CT, but the vital capacity was improved. The improvement of the respiratory muscle strength might reduce the respiratory symptom and increase the daily activity in this case. It was possible that dose of steroid was tapered early with tacrolimus, and the side effect which accompanies long-term medication of steroids could be avoided. In polymyositis, tacrolimus is very useful medicine in the steroid-resistant case or in the case suffering from the side effect of steroids. There is possibility that tacrolimus becomes one of the choices of the treatment of polymyositis. PMID- 15782612 TI - [Tactile hallucinations induced by trihexyphenidyl in a patient with Parkinson's disease]. AB - We report a 71-year-old woman with tactile hallucinations induced by trihexyphenidyl. Six and a half months after starting trihexyphenidyl, this patient with Parkinson's disease complained of tactile hallucinations, mainly on her feet, at night while in bed. Four days after stopping the medication, the sensations disappeared. Although tactile hallucinations induced by trihexyphenidyl are rare, it should be noted that the condition can be induced by anti-Parkinson's disease drugs. PMID- 15782613 TI - [A proposal for diagnostic and clinical assessment criteria for Alzheimer's disease]. AB - To clarify the risk and associated genes of Alzheimer's disease by genome-wide screening, a Japanese study group was organized in 2000 under Yasuo Ihara, Tokyo University, supported by a Grant-in-Aid for Science Research on Priority Areas (C) -Advanced Brain Science Project from Ministry of Education, Culture, Sports, Science and Technology, Japan. This is the first Japanese consortium study under permission of the ethical committees of the enrolled institutes based on the ethics guidelines for human genome/gene analysis research, Ministry of Education, Culture, Sports, Science and Technology Ministry of Health, Labor and Welfare Ministry of Economy, Trade and Industry. In this project, 2,000 genome samples from patients with Alzheimer's disease, 2,000 control subjects, and 200 siblings affected with Alzheimer's disease are collected and analyzed. For this purpose, it is necessary to analyze samples from accurately diagnosed Alzheimer patients and controls using standard criteria for diagnosis and neuropsychological evaluation, which have been confirmed by an evidence-based studying a Japanese population. Here, we propose criteria for the diagnosis and clinical assessment of Alzheimer's disease. This proposal consists of a definition of Alzheimer's disease based on recent advances in research, diagnostic criteria based on DSM IV, NINCDS-ADRDA and ICD-10, exclusion criteria for other dementia disorders, routine and detailed tests for neuropsychological and laboratory evaluations, criteria for neuroimaging and biomarkers, definitive diagnostic criteria and classification of clinical subtypes. PMID- 15782614 TI - [Transmission of tuberculosis (II)]. AB - In the previous paper (Kekkaku. 2004; 79: 509-518), the author has made a review on (1) development of droplet nuclei infection theory, (2) experimental basis and (3) epidemiological evidence of droplet nuclei infection, and has discussed about the factors influencing on tuberculosis infection. Various mode of transmission of tubercle bacilli from nonpulmonary sources was reviewed in the present paper, such as (1) infection from cutaneous tuberculosis, (2) infection in the bacteriology laboratory, (3) infection by needle stick, (4) transmission by bronchoscopic examination, (5) tuberculosis infection in the autopsy room, (6) infection followed vaccination and/or cortico-steroid injection therapy, (7) congenital tuberculosis, and (8) other rare transmission of tuberculosis. Moreover, three topics concerning tuberculosis infection were discussed, they are (1) tuberculosis infection risk index, (2) highly infectious case, and (3) virulence and infection. Infection risk index is the product of smear positivity (expressed by Gaffky Grading) by duration of cough (expressed by month). This index is being used widely at the occasion of contact survey at present in Japan so that instructions for use were discussed. PMID- 15782615 TI - [Validity of measuring time to detect growth of M. tbc by BACTEC MGIT960 system for quantitation and prediction of mycobacterial growth]. AB - STUDY DESIGN: Time to detect growth of M. tbc by BACTEC MGIT960 system was examined in sputum specimens collected from 114 patients with active pulmonary tuberculosis before and during antituberculosis therapy. By measuring TTD under chemotherapy, we tried to quantify mycobacterial growth and determine the sensitivity of MGIT system. RESULTS: The mean TTD significantly decreased in response to an increment in the range of the quantitation scale for solid media. Moreover, the TTD negatively correlated with colony counts (rho = - 0.636, P < 0.01). When automated monitoring continued until Day 28 after incubation, MGIT system had been capable of detecting 98% of Ogawa-positive specimens. The receiver operating characteristic (ROC) curve was plotted to determine the sensitivity and specificity in MGIT system, indicating the sensitivity of 98.3% corresponding cutoff level for TTD of Day 28. CONCLUSION: Measuring TTD in MGIT system could allow estimating the mycobacterial growth in similarly quantitative manner. The appropriate endpoint of monitoring could be decided as 4 weeks, accurately reflecting an outcome of cultivation with solid media. PMID- 15782616 TI - [Clinical evaluation of causes of death in patients with pulmonary tuberculosis]. AB - PURPOSE: We evaluated the cause of death in patients with active tuberculosis. OBJECT AND METHOD: 40 patients (male 32, female 8, mean age 76 years old), died under treatment for tuberculosis during 1999 to 2002 in our hospital, were analized. We compared patients' backgrounds, complications, extent of pulmonary tuberculosis, and outcome of the treatment between 40 died cases and 162 patients who were supposed to be successfully treated by cohort analysis (control group). RESULTS: 17 cases died of tuberculosis and 23 cases died of non-tuberculous diseases. Pneumonia occupied the top (9 cases) in the latter group. Died cases were higher in age and showed larger performance status score than the control group. Further, laboratory findings revealed poorer nutritional conditions and higher inflammatory reactions in the died group. The duration of symptoms before admission seemed to be longer in the died group, however, the difference with the control was not significant. All of the died group had complications and the rate of having liver diseases and cerebrovascular diseases was higher than that of the control group. Furthermore, the extent of lung lesions was more extensive in the died group. The frequency of changing drugs due to side effects was higher in the died group, therefore, the proportion of cases completed the standard treatment was lower in the died group. DISCUSSION AND CONCLUSION: Impossibility of continuing the standard treatment gave unfavorable impact in the died cases rather than the delay in their admission. Due to the above reasons, negative conversion of their sputum culture was difficult, and they died of tuberculosis directly or indirectly. Some of the patients who died of pneumonia (non tuberculous death) might be included in cases died of tuberculosis. PMID- 15782617 TI - [Process of acquiring drug resistance: retrospective review of records of MDR TB]. AB - PURPOSE: To investigate the reasons of acquiring drug resistance among MDR TB cases and to learn lessons for the prevention of acquiring of drug resistance. METHOD: Retrospective review of 159 MDR TB cases who were treated at Fukujuji Hospital from 1990 January to 2003 August. RESULT: We found that among 159 cases, 48 cases were infected with multidrug resistant M. tuberculosis bacilli, 35 cases acquired drug resistance, 7 cases were with the history of tuberculosis treatment before 1970 only, and that remaining 69 cases were difficult to evaluate because of the lack of informations on previous drug susceptibility tests. Among 35 cases that acquired drug resistance, the drug susceptibility test patterns before becoming MDR TB were categorized as follows: 12 HR susceptible, 18 H resistant R susceptible, 3 R susceptible (H unknown), and 2 H susceptible R resistant. The factors that may have influenced to acquire MDR were lack of modification of the regimen after knowing drug resistance among H resistant R susceptible cases, and defaulting among cases that were not evaluated (15/69) and H resistant R susceptible cases (3/18). DISCUSSION: Control of MDR TB needs to be strengthened. Proper drug susceptibility test, proper choice of drugs at the beginning of treatment and modification of treatment after knowing drug susceptibility test results are important for the prevention of MDR TB. Ensuring patient adherence to treatment is important in the medical institutions where drug susceptibility test is not properly done, in particular, for H resistant R susceptible cases, and guidance to these institutions by the public health centers should be intensified. PMID- 15782618 TI - [Basic characteristics of a novel diagnostic method (QuantiFERON TB-2G) for latent tuberculosis infection with the use of Mycobacterium tuberculosis-specific antigens, ESAT-6 and CFP-10]. AB - PURPOSES: To determine the optimum cut-off level of a newly developed method for diagnosing tuberculosis infection based on whole-blood interferon-gamma measurement, and to study the basic characteristics of the method. STUDY SUBJECTS: 1) A total of 220 young, healthy individuals having no apparent exposure to tuberculosis infection, most of whom have had a vaccination with BCG vaccine. 2) One hundred eighteen tuberculosis patients who were diagnosed by positive Mycobacterium tuberculosis on culture. 3) A group of 75 youngsters exposed to an infectious tuberculosis patient and who showed a strong tuberculin reaction (with erythema diameter of 30 mm or more). METHOD: Whole-blood specimens of donors were stimulated with antigens, i.e., ESAT-6 and CFP-10, and then cultured. Plasma concentrations of interferon-gamma discharged were then determined with QuantiFERON-CMI. Correlation between interferon-gamma concentrations in response to ESAT-6 and CFP-10, and their correlation with Mantoux test results were analyzed for various categories of donors. The Receiver Operating Characteristics analysis was performed considering the loss due to misclassification. [ RESULTS AND DISCUSSION: The optimum cut-off level was determined as 0.35 IU/ml for both ESAT-6 and CFP-10. This gave the test a sensitivity of 89.0% and specificity of 98.1% in detecting tuberculosis infection. The correlation of interferon-gamma response with tuberculin tests among BCG-vaccinated individuals was low, which suggested that the test was not influenced by previous BCG vaccination. The low correlation between ESAT-6 and CFP-10 tests suggested that the simultaneous use of the two tests was beneficial. As in the case of clinical tests in general, the cut-off should be set at a lower level when the test is applied to high prevalence situation and vice versa. PMID- 15782619 TI - [Clinical study on development of nontuberculous mycobacterial lung disease]. AB - DEVELOPEMENT OF MAC LUNG DISEASE: An increase of nodular bronchiectatic type of MAC lung disease becomes a problem among respiratory physician today. The reason is still unknown, but it seems to be globally recognized that this type of MAC disease is developing particularly in middle-aged woman. Some papers mentioned the existence of such type of MAC lung disease already early in the 70s, in Japan. Yamamoto described that 17 cases of middle lobe type lung disease out of 154 non-photochoromogen cases, and 76.5% were female, in 1970. Shimoide also pointed such type of 39 cases out of 240 MAC lung disease and 84.6% were female, in 1980. Prince reported MAC lung disease seen in old and middle age female of 21 cases including lethality example of 4 cases without a precedent disease in 1989. After his report, the international consensus of this peculiar type of MAC lung disease seems to be spread. In 1989, we compared 72 cases of nodular bronchiectatic type of MAC lung disease and 56 cases of diffuse panbronchiolitis (DPB) that was a most typical chronic airway disease at that time in Japan. The average age of disease onset of DPB group was 37.0 +/- 16.3 years old and that of MAC group was 54.5 +/- 16.3 years old. The percentage of female was 32% in DPB group and 87.5% in MAC group. It was highly possible that two groups belong different parent population. We could grasp that nodular bronchiectatic type of MAC lung disease patients is a unique group. We observed the serial films of 21 cases of nodular bronchiectatic MAC lung disease, and divide the progression of the disease to sequential 7 steps as Fig. 1. Small nodules progress to cavities in mean about 10 years. However, why is MAC which is opportunistic pathogen with weak virulence, able to form a lesion at unimpaired lung parenchyma? Is there really normal site? Why dose it start from lingula? Why is MAC seen a lot in woman? While it is extremely pathognomonic clinical picture, and, is an extremely interesting problem, most are still unidentified. STUDY OF MAC LUNG DISEASE TREATMENT: It was known that Mycobacterium kansasii lung disease is healed with a chemotherapy like analog of anti-tuberculosis chemotherapy, already in those days. However, the results of MAC lung disease chemotherapy were extremely poor. We tried to express a physicians experience quantitatively as follows, in 1987. The results of 8 weeks sputum culture on Ogawa egg medium were converted semi quantitatively to CFU numbers based on "Japanese standard guideline of Mycobacterium tuberculosis inspection". We exhibit the ratio of post-treatment consecutive 6 months culture yield to pre-treatment culture yield as response rate, about 110 pulmonary MAC cases. Through this study, we clarify the followings. The results of chemotherapy do not correlate susceptibility test for Mycobacterium tuberculosis. Multidrug regimen is more useful. Small extent of lesion is more responsive. Combination with aminoglycoside chemotherapy is more effective. These conclusions were almost same as the ATS guideline of 1990. New drugs such as, new macrolides and new quinolones appeared for pulmonary MAC treatment through the feedback from systemic MAC complicated AIDS treatments from the latter half of 90's. We measured the sensitive strain ratio at 2 mcg/ml of OFLX, CPFX, LVFX about 990 clinical isolates and could expect availability for M. kansasii or M. fortuitum, but these new quinolones are not enough effective for MAC. Also we examined MIC for various antimycobacterial agent by 50 MAC clinical isolates, and we could expect a certain availability of SPFX, GFLX, CPFX, CAM for MAC. The availability of clarithromycin (CAM) has been established through many randomized clinical trials for disseminated MAC complicated AIDS, but for pulmonary MAC, complete cure is still difficult if we use CAM including regimen. We performed surgical treatment for relatively young patients with localized lesions. We carry out the adaptation reference such as Table, now. The localization of the lesions become a problem at surgical resection. Through the study of our 55 surgical treatment cases, 8 cases (67%) relapsed out of 12 cases which had destructive airway structure in unresected lung field. On the other, only 1 case relapsed (10%) relapsed out of 10 cases without airway destruction in unresected lung. Therefore, even if there is a little dispersal focus without airway destruction in the other pulmonary lobe except purpose focus of resection, it seems that control is possible by post operational chemotherapy. LONG SURVIVAL: As overall consequence, we calculate the survival curves of 201 pulmonary MAC patients visited Tokyo National Hospital from 1953. The survival medium value was 7332 days. The prognosis of nodular bronchiectatic type was better than that of post-tuberculosis type. Extent of disease measured by chest X ray examination at the time of first visit may be a most affecting factor to the survival rate. PMID- 15782620 TI - [Nursing of tuberculosis patients--quality nursing to support patient's compliance to treatment]. AB - Tuberculosis control program in Japan focuses more on completion of treatment. The activities for patients to complete treatment are being actively done with collaboration between nurses of hospitals and public health nurses of the health centers. In 2000, Ministry of Health Welfare announced DOTS program version Japan. As a result, health centers of big cities implemented DOTS for homeless tuberculosis patients and hospitals DOT for in-patients. In 2003, the government demonstrated the scheme of DOTS strategy Expansion Program version Japan, which includes community DOTS types to be selected depend upon the risk of default with an individual patient. It is necessary to develop and utilize social and human resources in the community to expand surely supporting system for patient's compliance. Mutual understanding and collaboration of the relevant organizations become very important. In this symposium, four panels from hospital, clinic and public health center discuss on the current situation and challenge of supporting system and the assessment of treatment outcome. 1. DOTS implementation with collaboration on nursing activities between hospital and public health center in Kyoto Prefecture: Ikuyo HIROHATA (National Hospital Organization Minami Kyoto National Hospital). 2. From standpoint of clinical practice: Hidenori MASUYAMA (Japan Anti-Tuberculosis Association Shibuya Clinic). 3. The activities for case support based on DOTS Program in Wakayama Prefecture: Kimiko KAWASAKI (Tanabe Public Health Center, Wakayama Prefecture). 4. Assessment of supporting activities for patient's compliance: Tomoko TAKANO (Uki Public Health Center, Kumamoto Prefecture). Three speakers gave additional comments and advice on quality supporting and nursing activities for patient's compliance through good coordination between hospitals and publics health centers. PMID- 15782621 TI - Rapid viral diagnosis: role of electron microscopy. AB - Starting in the 1960, electron microscopy (EM) became widely applied also in viral diagnosis. During the 1970th and 80th, many new agents were characterized from diagnostic cell cultures and clinical specimens. The wide introduction of ELISA- and PCR-techniques as well as cost-arguments recently reduced the role of EM in routine viral diagnosis. Compared to other diagnostic techniques, however, EM excells by speed and "open view", i.e. by the ability to detect also the "un expected" without the need for specific reagents. As shown in 2003 by the elucidation of the SARS pandemia and the human monkeypox outbreak in US, EM is well suited as a safe, front-line diagnostic method in infectious diseases emergencies and/or in possible bioterrorist attacks. PMID- 15782622 TI - Predictive value of early virological response to treatment with different interferon-based regimens plus ribavirin in patients with chronic hepatitis C. AB - Early virological response (EVR) to different interferon-based regimens plus ribavirin and its ability to predict the outcome of therapy in patients with chronic hepatitis C were investigated. The study design was as follows: 64 naive patients were considered, 32/64 received pegylated interferon alpha-2b (Peg-IFN alpha2b) plus ribavirin and the remaining 32 received leucocyte interferon alpha (IFN-alpha) plus ribavirin. At week 4 of treatment, EVR was present in 68.7% and 37.5% of patients treated with Peg-IFN-alpha2b plus ribavirin, and with leucocyte interferon alpha (IFN-alpha) plus ribavirin, respectively (p = 0.024). At week 12, the cumulative EVR rates did not differ between the two groups (71.9% vs 56.2%, p >0.05) because a higher proportion of patients achieved EVR for the first time after more than 4 weeks of therapy in the standard IFN-alpha group. Sustained virological response (SVR) rates, however, resulted significantly higher in the Peg-IFN-alpha2b group (65.6% vs 37.5%; p = 0.045) since a higher proportion of patients who received standard IFN-alpha relapsed during the follow up. In the standard IFN-alpha group, HCV genotype 1 (p = 0.035), high baseline viral load (p = 0.035) and the presence of bridging fibrosis/cirrhosis (p = 0.011) were closely associated with significantly lower SVR rates. In the Peg-IFN alpha2b group, only bridging fibrosis/cirrhosis (p = 0.02) negatively influenced the outcome of treatment. Overall, 33/41 (80.5%) patients with EVR at week 12 were sustained responders, yielding a positive predictive value (PPV) of 0.80. However, when SVR was related to the time taken to reach EVR, 32/34 (94.1%) patients with EVR at week 4 of therapy (PPV = 0.94) versus 1/7 (14.3%) patients who had EVR after more than 4 weeks of therapy (PPV = 0.14) resulted sustained responders (p = 0.000057). In conclusion, EVR at week 4 of treatment is strongly associated with the likelihood of achieving SVR, regardless of the therapeutic regimen. However, when compared with standard IFN-alpha plus ribavirin, treatment with Peg-IFN-alpha2b plus ribavirin significantly increases the probability of viral clearance within the first 4 weeks of treatment. Finally, patients who do not clear the virus within the first 12 weeks of treatment have no chance of achieving SVR, justifying discontinuation of therapy in these patients. PMID- 15782623 TI - Treatment of genotype 1b HCV-related chronic hepatitis: efficacy and toxicity of three different interferon alfa-2b/ribavirin combined regimens in naive patients. AB - This prospective open-label randomized trial of chronic hepatitis C genotype-1b patients compared compared the efficacy and safety of peg-interferon alfa-2b administered once-weekly versus interferon alfa-2b thrice-weekly or daily, both in combination with ribavirin. Seventy-eight previously untreated patients, with biopsy-documented genotype 1 chronic HCV and persistently elevated ALT levels and detectable HCV RNA, were randomized (26 subjects each) to receive: interferon alfa-2b at 6MIUs.c./three-times-weekly (group A) or interferon alfa-2b, 3MIUs.c./daily (group B) or peg-interferon alfa-2b 1.5mcg/Kg s.c./once-weekly (group C). All regimens included standard weight-based doses of ribavirin (800, 1,000 or 1,200 mg/day) administered for 52-weeks. Patients in the three groups were comparable for age, sex, viral load, ALT value and histological-activity index (HAI). Therapy was completed by 22, 20 and 23 patients in groups A, B and C, respectively. At the end of treatment, a complete (biochemical and virological) response was observed in 50.0% patients of group A, 57.7% of group B and 65.4% of group C. After an additional 24-weeks of follow-up, a sustained response was observed in 26.9%, 46.1% and 50.0% of patients in groups A, B or C, respectively. Therapy was discontinued by 4, 6 and 2 patients because of adverse events in the above three groups. In naive patients with chronic genotype-lb hepatitis C, a 48 week therapy with peg-interferon or interferon at daily doses combined with ribavirin were both more effective than treatment with thrice weekly interferon in inducing end of treatment and sustained response. Peg interferon treatment was better tolerated and provoked significantly fewer therapy discontinuations. PMID- 15782624 TI - Detection of influenza A virus by RT-PCR and standard methods in experimental infection of Ducks. AB - Cloacal swabs collected from mallard ducks (Anas platyrhynchos) experimentally infected with a H7N1 avian influenza strain were examined by Reverse Transcription Polymerase Chain Reaction to detect the influenza A virus. Reverse Transcription Polymerase Chain Reaction was compared with standard methods: inoculation of embryonated chicken eggs and inoculation of three established cell lines: Newborn Swine Kidney cells, Newborn Pig Trachea cells and Madine Darby Canine Kidney cells. Reverse Transcription Polymerase Chain Reaction was performed using a set of primers based on conserved regions of the matrix and nucleoprotein genes. PMID- 15782625 TI - Comparative evaluation of two enzyme linked immunosorbent assay methods and three Western Blot methods for the diagnosis of culture-confirmed early Lyme borreliosis in Italy. AB - This study investigated the onset and development of the immune response to Borrelia burgdorferi infection in 30 Italian patients with culture-confirmed Lyme Borreliosis in the stage of erythema migrans (EM). All patients received antimicrobial treatment when entering the study and were prospectively evaluated monthly for up to 30 days after enrolment. A total of 60 serially collected serum samples were tested by using two different commercial enzyme-linked immunosorbent assays (ELISAs): Anti-Borrelia plus VlsE ELISA, Euroimmun, and the synthetic peptide-based ELISA, Quick ELISA C6, Immunetics. Sixty-five potentially cross reacting sera were also tested. Anti-Borrelia plus VlsE ELISA IgG was far more sensitive than Quick ELISA C6 (56.6% and 33.3%, respectively). Moreover, considering that 17 additional sera from the first bleeding group of Lyme disease patients were IgM positive when tested by Anti-Borrelia plus VlsE IgM, the sensitivity of Anti-Borrelia plus VlsE as a whole system rose to 85.0%. Nevertheless, due to the specificity values of Anti-Borrelia plus VlsE ELISA identified in this study (98.5% for IgG and 78.5% for IgM), the need of a confirmatory test for the diagnosis of Lyme disease remains. All the sera were also tested by two different commercial Western Blot (WB) assays: Euroline-WB against Borrelia, Euroimmun, and Qualicode B. burgdorferi WB, Immunetics, in comparison with a multispecies "home made" WB. Performances of the three WB methods for the detection of IgM were very similar. On the contrary, these WBs performed with different values of sensitivity and specificity when IgGs were evaluated. The most sensitive method was the "home-made" WB IgG (71.7%), followed by the Euroline-WB IgG against Borrelia (68.3%). Qualicode B. burgdorferi WB IgG demonstrated to be only 26.6% sensitive. Both "home-made" WB IgG and Qualicode B. burgdorferi WB IgG were 100% specific, whereas Euroline-WB IgG against Borrelia scored 12 cross-reacting samples as borderline, showing a specificity value of 80.0%. PMID- 15782626 TI - Biliary tract disease as a risk factor for Plesiomonas shigelloides bacteraemia: a nine-year experience in a Hong Kong hospital and review of the literature. AB - We report the epidemiology, clinical disease spectrum, treatment, and outcome of Plesiomonas shigelloides bacteraemia in our hospital over a nine-year period and compare the characteristics of patients with P. shigelloides bacteraemia in our hospital with those reported in the literature. During the nine-year period (1995 -2003), a total of seven patients developed P. shigelloides bacteraemia. All cases occurred during the late spring, summer, and early autumn (May-October). All patients were over 75 and had underlying diseases (biliary tract diseases in four and malignancies in three). Six had acute cholangitis and five had polymicrobial bacteraemia. Overall two patients died. Compared with patients with P. shigelloides bacteraemia reported in the literature, patients with P. shigelloides bacteraemia in our hospital were associated with old age (P<0.001), underlying biliary tract diseases (P<0.001), acute cholangitis (P<0.001), and polymicrobial bacteraemia (P<0.001). There are major geographical differences of disease association in P. shigelloides bacteraemia. The sources of P. shigelloides in our patients were probably in the gastrointestinal tract. PMID- 15782627 TI - Characterization of methicillin resistant Staphylococcus aureus (MRSA) isolated at the Policlinico Hospital of Bari (Italy). AB - Methicillin resistant Staphylococcus aureus (MRSA) is an emerging problem. We studied 71 MRSA strains for the presence of mecA gene by PCR, for the enterotoxins production and susceptibility to antimicrobials. In addition, the suitability of Random Amplified Polymorphic DNA analysis (RAPD) and Hypervariable Region (HVR)--PCR as molecular tools for typing MRSA was also tested. All the 71 strains previously found MRSA with conventional methods, presented the gene mec A. By molecular typing five distinct amplicons were found. MRSA with two DRUS were the most common type. RAPD analysis clustered MRSA in 8 groups, three of which were the most common. 26.8% of MRSA produce enterotoxins with a prevalence of type A. MRSA exhibited resistance to all quinolones tested and to gentamycin. Our data suggest that a typing method based on RAPD combined with HVR-PCR may be useful to compare MRSA isolated in a hospital environment, whereas PFGE may be used for further analysis. PMID- 15782628 TI - Direct detection of Mycobacterium tuberculosis complex in pulmonary and extrapulmonary samples by BDProbeTec ET system. AB - BDProbeTec ET (Becton Dickinson, Sparks, Md, USA) is a fully automated walkaway system based on strand displacement amplification (SDA) technology that provides a method for the direct detection of Mycobacterium tuberculosis complex (MTBC) target sequence. The purpose of this study was to evaluate the ability of BDProbeTec ET system to detect MTBC directly from clinical specimens and compare the results with staining and culture. From February 2002 through December 2003 a total of 1521 [pulmonary (n=1329) and extrapulmonary (n=192)] specimens from 1518 patients were examined by BDProbeTec ET system for the detection of MTBC and the results were compared to those obtained by microscopy and liquid culture (BACTEC 9000 MB, Becton Dickinson). MTBC was cultivated from 65 specimens (60 pulmonary and 5 extrapulmonary) of which 43 (66.2%) (42 pulmonary and 1 extrapulmonary) were smear positive and 22 (33.8%) (18 pulmonary and 4 extrapulmonary) were smear negative. BDProbeTec ET detected MTBC in 58 (55 pulmonary and 3 extrapulmonary) of the 65 culture-positive specimens. Although the BDProbeTec ET system gave five false-negative results among the 18 smear-negative culture-positive pulmonary specimens, our results demonstrate that the BDProbeTec ET system is a reliable tool in smear-positive samples and given its technical characteristics it can be used for the rapid detection of MTBC in either pulmonary or extrapulmonary samples. PMID- 15782629 TI - Biotypes and randomly amplified polymorphic DNA (RAPD) profiles of subgingival Candida albicans isolates in HIV infection. AB - A group of subgingival isolates of C. albicans recovered from Italian HIV positive (HIV+) subjects were characterized both phenotypically and genotypically. Phenotyping of the isolates was carried out by a biotyping method based on the enzyme profiles, carbohydrate assimilation patterns and boric acid resistance of the yeasts. Genotyping was performed through randomly amplified polymorphic DNA (RAPD) analysis. Five biotypes were found among the 29 subgingival C. albicans strains examined. The predominant biotypes were A1R (55.17%), A1S (24.14%), and A2R (13.79%), while the biotypes A11R and A13R were represented by a single isolate each. RAPD profiles identified 15 genotypes among the 29 isolates. Almost every individual harboured his/her own specific isolate and in three out of the six subjects with multiple isolates (two to six each) more than one genotype (two to six) was found. The biotype distribution we found is consistent with previous reports on C. albicans isolates from other oral sources, whereas the resistance to boric acid was highly frequent in subgingival strains. RAPD analysis showed high genetic heterogeneity within subgingival isolates, also when isolates were phenotypically identical. These findings, obtained from HIV+ subjects living in Southern Italy, may be useful as baseline information on subgingival C. albicans colonization in the Mediterranean area. PMID- 15782630 TI - Evaluation of chromogenic medium (ORSAB) for routine identification of methicillin resistant Staphylococcus aureus (MRSA). AB - Performance of chromogenic medium (ORSAB) for routine detection of methicillin resistant S. aureus (MRSA) was evaluated on 510 specimens collected from patients suspected of MRSA infection or colonization. Addition of ORSAB plates to the routine protocol allowed MRSA identification in 24 hours from samples plating. In 18 samples MRSA colonies were identified only on ORSAB plates, those cases would have been missed by routine protocol alone. PMID- 15782631 TI - A case of granulomatous skin lesions caused by Mycobacterium marinum in the Campania region. AB - The diagnosis of cutaneous Mycobacterium marinum infection is frequently presumptive, as detection by conventional methods is difficult. We describe a patient with granulomatous skin lesions on the right dorsal hand and forearm. Histological examinations were presumptive for mycobacterium lesions. We identified Mycobacterium marinum directly in the patient's lesional skin biopsy combining polymerase chain reaction (PCR) amplification using Mycobacterium genus specific primers, and subsequent restriction enzyme analysis enabling identification to the species level. The symptoms were no longer present after specific therapy, thereby confirming the initial diagnosis. PMID- 15782632 TI - Differences in intensity and specificity of hypersensitive response induction in Nicotiana spp. by INF1, INF2A, and INF2B of Phytophthora infestans. AB - Elicitins form a family of structurally related proteins that induce the hypersensitive response (HR) in plants, particularly Nicotiana spp. The elicitin family is composed of several classes. Most species of the plant-pathogenic oomycete genus Phytophthora produce the well-characterized 10-kDa canonical elicitins (class I), such as INF1 of the potato and tomato pathogen Phytophthora infestans. Two genes, inf2A and inf2B, encoding a distinct class (class III) of elicitin-like proteins, also occur in P. infestans. Unlike secreted class I elicitins, class III elicitins are thought to be cell-surface-anchored polypeptides. Molecular characterization of the inf2 genes indicated that they are widespread in Phytophthora spp. and occur as a small gene family. In addition, Southern blot and Northern blot hybridizations using gene-specific probes showed that inf2A and inf2B genes and transcripts can be detected in 17 different P. infestans isolates. Functional secreted expression in plant cells of the elicitin domain of the infl and inf2 genes was conducted using a binary Potato virus X (PVX) vector (agroinfection) and Agrobacterium tumefaciens transient transformation assays (agroinfiltration), and resulted in HR-like necrotic symptoms and induction of defense response genes in tobacco. However, comparative analyses of elicitor activity of INF1, INF2A, and INF2B revealed significant differences in intensity, specificity, and consistency of HR induction. Whereas INF1 induced the HR in Nicotiana benthamiana, INF2A induced weak symptoms and INF2B induced no symptoms on this plant. Nonetheless, similar to INF1, HR induction by INF2A in N. benthamiana required the ubiquitin ligase associated protein SGT1. Overall, these results suggest that variation in the resistance of Nicotiana spp. to P. infestans is shadowed by variation in the response to INF elicitins. The ability of tobacco, but not N. benthamiana, to respond to INF2B could explain differences in resistance to P. infestans observed for these two species. PMID- 15782634 TI - The T-DNA oncogene A4-orf8 from Agrobacterium rhizogenes A4 induces abnormal growth in tobacco. AB - The related orf8 and iaaM T-DNA genes from Agrobacterium are each composed of two distinct parts. The 5' parts (called Norf8 or NiaaM) encode a 200-amino-acid (aa) sequence with homology to various T-DNA oncoproteins such as RolB, RolC, and 6b. The 3' parts (Corf8 or CiaaM) encode a 550-aa sequence with homology to IaaM proteins from Pseudomonas and Pantoea spp. Whereas iaaM genes encode flavin adenine dinucleotide (FAD)-dependent tryptophan 2-monooxygenases that catalyze the synthesis of indole-3-acetamide (IAM), A4-orf8 from Agrobacterium rhizogenes A4 does not. Plants expressing a 2x35S-A4-Norf8 construct accumulate soluble sugars and starch. We now have regenerated plants that express the full-size 2x35S-A4-orf8 and the truncated 2x35S-A4-Corf8 gene. 2x35S-A4-Corf8 plants accumulate starch and show reduced growth like 2x35S-A4-Norf8 plants but, in addition, display a novel set of characteristic growth modifications. These consist of leaf hypertrophy and hyperplasia (blisters); thick, dark-green leaves; thick stems; and swollen midveins. Mutations in the putative FAD-binding site of A4-Orf8 did not affect the blister syndrome. Plants expressing 2x35S-A4-Corf8 had a normal phenotype but contained less starch and soluble sugars than did wild type plants. When 2x35S-A4-Corf8 plants were crossed to starch-accumulating 2x35S A4-Norf8 plants with reduced growth, A4-Corf8 partially restored growth and reduced starch accumulation. A4-Corf8xA4-Norf8 crosses did not lead to the blister syndrome, suggesting that this requires physical linkage of the A4-NOrf8 and A4-COrf8 sequences. PMID- 15782633 TI - Evidence that RNA silencing-mediated resistance to beet necrotic yellow vein virus is less effective in roots than in leaves. AB - In plants, RNA silencing is part of a defense mechanism against virus infection but there is little information as to whether RNA silencing-mediated resistance functions similarly in roots and leaves. We have obtained transgenic Nicotiana benthamiana plants encoding the coat protein readthrough domain open reading frame (54 kDa) of Beet necrotic yellow vein virus (BNYVV), which either showed a highly resistant or a recovery phenotype following foliar rub-inoculation with BNYVV. These phenotypes were associated with an RNA silencing mechanism. Roots of the resistant plants that were immune to foliar rub-inoculation with BNYVV could be infected by viruliferous zoospores of the vector fungus Polymyxa betae, although virus multiplication was greatly limited. In addition, virus titer was reduced in symptomless leaves of the plants showing the recovery phenotype, but it was high in roots of the same plants. Compared with leaves of silenced plants, higher levels of transgene mRNAs and lower levels of transgene-derived small interfering RNAs (siRNAs) accumulated in roots. Similarly, in nontransgenic plants inoculated with BNYVV, accumulation level of viral RNA-derived siRNAs in roots was lower than in leaves. These results indicate that the RNA silencing mediated resistance to BNYVV is less effective in roots than in leaves. PMID- 15782635 TI - The plant gene CCD1 selectively blocks cell death during the hypersensitive response to Cauliflower mosaic virus infection. AB - The P6 protein of Cauliflower mosaic virus (CaMV) W260 elicits a hypersensitive response (HR) on inoculated leaves of Nicotiana edwardsonii. This defense response, common to many plant pathogens, has two key characteristics, cell death within the initially infected tissues and restriction of the pathogen to this area. We present evidence that a plant gene designated CCD1, originally identified in N. bigelovii, can selectively block the cell death pathway during HR, whereas the resistance pathway against W260 remains intact. Suppression of cell death was evident not only macroscopically but also microscopically. The suppression of HR-mediated cell death was specific to CaMV, as Tobacco mosaic virus was able to elicit HR in the plants that contained CCD1. CCD1 also blocks the development of a systemic cell death symptom induced specifically by the P6 protein of W260 in N. clevelandii. Introgression of CCD1 from N. bigelovii into N. clevelandii blocked the development of systemic cell death in response to W260 infection but could not prevent systemic cell death induced by Tomato bushy stunt virus. Thus, CCD1 blocks both local and systemic cell death induced by P6 of W260 but does not act as a general suppressor of cell death induced by other plant viruses. Furthermore, experiments with CCD1 provide further evidence that cell death could be uncoupled from resistance in the HR of Nicotiana edwardsonii to CaMV W260. PMID- 15782636 TI - Recombination events generating a novel Rp1 race specificity. AB - Genes at the maize Rp1 rust resistance complex often mispair in meiosis, which allows genes to recombine unequally, creating recombinant haplotypes. Four recombinant haplotypes were identified from progeny of an Rp1-D/Rp1-I heterozygote that conferred a nonparental resistance specificity designated Rp1 I*. Sequence comparisons of paralogs in the recombinant and parental haplotypes demonstrated that all four recombinants were derived from intergenic (between gene) recombination events. The sequence of paralogs in the HRp1-I parental haplotype indicated this haplotype includes 41 or more rp1 genes, at least 31 of which are transcribed. The results indicate that most of the novel resistance specificities that have arisen spontaneously at Rp1 are the result of reassort ment of existing Rp1 genes. PMID- 15782638 TI - The Pseudomonas chlororaphis PCL1391 sigma regulator psrA represses the production of the antifungal metabolite phenazine-1-carboxamide. AB - The rhizobacterium Pseudomonas chlororaphis PCL1391 produces the antifungal metabolite phenazine-1-carboxamide (PCN), which is a crucial trait in its competition with the phytopathogenic fungus Fusarium oxysporum f. sp. radicis lycopersici in the rhizosphere. The expression of the PCN biosynthetic gene cluster in PCL1391 is population density-dependent and is regulated by the quorum sensing genes phzI and phzR via synthesis of the autoinducer N-hexanoyl-L homoserine lactone (C6-HSL). Here, we describe the identification of an additional regulatory gene of PCN biosynthesis in PCL1391. A mutation in the psrA gene (Pseudomonas sigma regulator), the gene product of which is a member of the TetR/AcrR family of transcriptional regulators, resulted in increased production of autoinducer molecules and PCN. Expression studies showed that inactivation of psrA resulted in increased expression of the phzI and phzR genes and the phz biosynthetic operon and that introduction of functional copies of psrA represses the expression of these genes, resulting in reduced production of autoinducer signal and PCN. Surprisingly, inactivation of psrA in the phzI or phzR quorum sensing mutants, which do not produce detectable amounts of PCN and autoinducers by themselves, restored PCN biosynthesis. This phenomenon was accompanied by the appearance of compounds with autoinducer activities migrating at the positions of C4-HSL and C6-HSL on C18 reverse phase-thin-layer chromatography. These observations indicate that PsrA also represses at least one silent, yet unidentified, quorum-sensing system or autoinducer biosynthetic pathway in PCL1391. The expression of psrA declines at the onset of the stationary phase at the same moment at which quorum-sensing (-regulated) genes are activated. In addition, expression studies in a psrA- and a multicopy psrA background showed that psrA is autoregulated. Multiple copies of psrA repress its own expression. Mutation of gacS, encoding the sensor kinase member of a two-component global regulatory system significantly reduced production of autoinducers and PCN. We show a novel link between global regulation and quorum sensing via the PsrA regulator. PMID- 15782639 TI - Glutathione and homoglutathione play a critical role in the nodulation process of Medicago truncatula. AB - Legumes form a symbiotic interaction with bacteria of the Rhizobiaceae family to produce nitrogen-fixing root nodules under nitrogen-limiting conditions. This process involves the recognition of the bacterial Nod factors by the plant which mediates the entry of the bacteria into the root and nodule organogenesis. We have examined the importance of the low molecular weight thiols, glutathione (GSH) and homoglutathione (hGSH), during the nodulation process in the model legume Medicago truncatula. Using both buthionine sulfoximine, a specific inhibitor of GSH and hGSH synthesis, and transgenic roots expressing GSH synthetase and hGSH synthetase in an antisense orientation, we showed that deficiency in GSH and hGSH synthesis inhibited the formation of the root nodules. This inhibition was not correlated to a modification in the number of infection events or to a change in the expression of the Rhizobium sp.-induced peroxidase rip1, indicating that the low level of GSH or hGSH did not alter the first steps of the infection process. In contrast, a strong diminution in the number of nascent nodules and in the expression of the early nodulin genes, Mtenod12 and Mtenod40, were observed in GSH and hGSH-depleted plants. In conclusion, GSH and hGSH appear to be essential for proper development of the root nodules during the symbiotic interaction. PMID- 15782641 TI - [Personal experience in the diagnosis of vestibular patients: transition from empirism to scientific vestibular medicine]. PMID- 15782640 TI - RpoN (sigma54) controls production of antifungal compounds and biocontrol activity in Pseudomonas fluorescens CHA0. AB - Pseudomonas fluorescens CHA0 is an effective biocontrol agent of root diseases caused by fungal pathogens. The strain produces the antibiotics 2,4 diacetylphloroglucinol (DAPG) and pyoluteorin (PLT) that make essential contributions to pathogen suppression. This study focused on the role of the sigma factor RpoN (sigma54) in regulation of antibiotic production and biocontrol activity in P. fluorescens. An rpoN in-frame-deletion mutant of CHAO had a delayed growth, was impaired in the utilization of several carbon and nitrogen sources, and was more sensitive to salt stress. The rpoN mutant was defective for flagella and displayed drastically reduced swimming and swarming motilities. Interestingly, the rpoN mutant showed a severalfold enhanced production of DAPG and expression of the biosynthetic gene phlA compared with the wild type and the mutant complemented with monocopy rpoN+. By contrast, loss of RpoN function resulted in markedly lowered PLT production and plt gene expression, suggesting that RpoN controls the balance of the two antibiotics in strain CHA0. In natural soil microcosms, the rpoN mutant was less effective in protecting cucumber from a root rot caused by Pythium ultimum. Remarkably, the mutant was not significantly impaired in its root colonization capacity, even at early stages of root infection by Pythium spp. Taken together, our results establish RpoN for the first time as a major regulator of biocontrol activity in Pseudomonas fluorescens. PMID- 15782642 TI - [Neonatal hearing loss screening: our results three years after starting the program]. AB - Since the 90s there increasing interest in early detection of neonatal hearing loss due to the feasibility of universal neonatal screening as well as to the possibility of early intervention. This has led to set up neonatal screening programs based upon otoacoustic emissions and to a great increase in the number of publications in the field. Our hospital started a neonatal hearing loss screening program using otoacoustic emission in March 2000. We used a Capella Cochlear Emissions (MADSEN electronics) equipment. Until May 2003 we have performed the test in 6372 children with a median age of 71 days. In our study, the sensibility of the test was 90.9% and the specificity 99.7%. Our results are similar to those reported in the literature although the age of the children was higher and we have found an incidence of hearing loss lower than other studies. PMID- 15782643 TI - [Incidence of complications of otitis media]. AB - OBJECTIVE: Determine the incidence of otological and endocranial complications of acute otitis media, taking into account previous treatments and the development of complications. MATERIAL AND METHODS: Retrospective follow-up study between March 1996-2003 including 16 patients: 9 men, 7 women. RESULTS: Nine patients (56.25%) had intracranial complications and seven otological ones. In the first group, 6 developed meningitis by Streptococcus pneumoniae, one sigmoid sinus thromboflebitis, one multiple abscesses, and one subdural empiema and encephalitis. Otological complications were six acute mastoiditis and the other one a facial paralysis. The child with thromboflebitis underwent mastoidectomy and closed meningeal comunication; 5 of the acute mastoiditis were operated on and the other received traditional treatment. The meningitis received medication. Two died, one because of a subdural empiema and the other due to meningitis. CONCLUSION: Despite the advances made by antibiotherapy and diagnosis, the complications of otitis media are still frequent. The pediatrician and otorhinolaryngologist should be on the alert for these as well as for Pneumococcus and its high resistance. PMID- 15782644 TI - [Statistical study and clinical results by comparison of two different protocols in the treatment of sudden hearing loss]. AB - Sudden deafness goes on being a clinical entity of unknown cause for which several theories and treatments have been proposed. We are reporting a comparative study of two different protocols that we have performed on 60 patients (divided in two groups of 30) diagnosed and admitted with sudden hearing loss between 1989 and 2003. The main goal is to prove if there is a statistical and significant difference among both, in respect of the audiologic improvement obtained, after two weeks of therapy at east. We have applied the normal approximation of Mann-Whitney's test and we can argue, after its result, our current protocol (piracetam, pentoxifiline, metilprednisolone), which includes 4 days of endovenous treatment another 10 days at home. The presence of vestibular symptoms darkens the prognosis since about 95% of the patients with them have not improved significantly (p<0.05). PMID- 15782645 TI - [Hearing loss associated to ulcerative colitis]. AB - OBJECTIVE: To value the eventual immunomediation in sensorineural hearing loss (SHL) on patients bearing of ulcerative colitis (UC). MATERIAL AND METHODS: In a group of forty-nine cases with a mean age of 41.6+/-9.3 years old we studied the hearing loss level, the disease activity index, the peripheral blood inflammation markers and the anticochlear antibodies by mean of Western-blot technique (WB). RESULTS: The 26.5% knew about their deafness, although SHL was detected in 59.1% of cases. The mean age of onset was 40.3+/-9.8 years. 48.9% showed a positive WB, always in 68-70 kDa molecular weight blots. Moreover, patients with positive WB showed more severe deafness, higher disease activity and more altered parameters, especially erythrosedimentation rate. CONCLUSIONS: Audiologic and peripheral blood findings observed allow us to establish a reasonable suspicion of an autoimmune or immunomediated pathway of hearing loss on UC. PMID- 15782646 TI - [Combined intratympanic and intravenous dexomethasone to control vertigo in Meniere disease]. AB - OBJECTIVE: To verify the result in the use of intratympanic corticosteroids, combined with intravenous, in patients with Meniere disease (MD). MATERIAL AND METHODS: Ten patients with vertigo due to MD in stages 4-5 or 6 were included in the study. Combined intratympanic and intravenous dexamethasone was infiltrated. The number of episodes among the months 18 and 24 of treatment, comparing them with the number of episodes 6 months before treatment (according to the norms of the American committee for the hearing and the equilibrium, 1995) are studied. RESULTS: All patients improved significantly. In one of them treatment was repeated after 6 months. A number of patients with hearing improvement were observed. CONCLUSIONS: The use of combined dexamethasone may be useful to control vertigo due to MD, to avoid or to be a prior step to other more aggressive treatments. PMID- 15782648 TI - [Idiopathic Frey's syndrome under the appearance of a recurrent otitis externa]. AB - Frey syndrome has been observed especially in patients who have undergone a parotidectomy operation, but also in zoster herpes, in parotiditis, condilea fractures, obstetric traumatisms with forceps and in surgery of the meningioma of the cerebellopontine angle. It also appears without previous surgery, like in our case. In these circumstances it is believed that a clinical neuritis, primary or secondary to a neighbouring inflammation may cause the start of this disorder. Several treatments have been suggested which highlights the difficulty of them. The most effective one is the intradermic injection of botulinum toxin type A. Its use in Frey's syndrome was initiated by Drobik and Laskawi in 1995. Since then the references to its use are numerous. Nevertheless, it is a treatment which has been introduced very few times in our country. PMID- 15782647 TI - [Influence of the nasogastric tube in the pathology of the maxillary sinus]. AB - INTRODUCTION: Nasoesophageal tube (NT) is very often used by the ear, nose and throat specialists in head and neck oncological surgery. It is well known the irritative effect that it has on the nasosinusal mucosa. The aim of this study is to analyze the relationship between the use of the NT and the pathology of the maxillary sinuses. MATERIAL AND METHOD: The study was carried out in 25 patients treated in our department who had been fed by NT, after surgery. A plain sinus X ray was performed in the pre-operatory period and a control radiograph in the post-operatory period, and the results have been analyzed. RESULTS AND CONCLUSIONS: No sinusitis had been observed whatever length of time the NT remained in situ. PMID- 15782649 TI - [Surgical repair of an anastomotic stenosis in pharyngeal reconstructions using antebrachial free flaps]. AB - To obtain suitable deglutoria function and less morbidity after hypopharyngeal surgery, antebrachial free flaps is a very useful option. A posible complication is oral feeding impossibility due to neopharyngeal stenosis. Tumor recurrence must be excluded. We described two cases of hypopharyngeal stenosis after antebrachial free flap reconstruction, proposing surgical reanastomosis with fibrous ring resection and salivary by-pass stent. PMID- 15782637 TI - Large-scale gene discovery in the oomycete Phytophthora infestans reveals likely components of phytopathogenicity shared with true fungi. PMID- 15782650 TI - Experts glimpse a brighter future for capitation in 2005. PMID- 15782651 TI - PacifiCare ordered to pay 6.9 million dollars to bankrupt IPA. AB - PacifiCare of Texas was ordered to pay a physician group nearly 7 million million for improperly terminating a capitated contract. The group's attorney believes the arbitrator's decision sends a message to health plans to contract fairly. PMID- 15782652 TI - Survey: capitated groups outperform fee-for-service physicians. AB - Capitated medical groups continue to earn more per physician and appear to be more productive in terms of relative value units than fee-for-service physicians, according to data from the American Medical Group Association's 2004 Medical Group Compensation & Financial Survey. PMID- 15782653 TI - Geisinger clinic moves away from risk as doctors move toward pay for performance. AB - Geisinger Clinic became the latest large group to drop capitation, but the reason was philosophical, not financial. The clinic's CFO acknowledges the physicians made money on their capitated business. PMID- 15782654 TI - Morphology of the lymph nodes in bottlenose dolphin (Tursiops truncatus) and striped dolphin (Stenella coeruleoalba) from the Adriatic Sea. AB - Morphology of the lymph nodes was examined in six bottlenose dolphins (Tursiops truncatus) and three striped dolphins (Stenella coeruleoalba) from the Adriatic Sea. All animals had been found dead in nature. One group of the nodes was taken from the tracheal branching area and was marked as bifurcational lymph node, and the other group was taken from the mesenteric root and was marked as mesenteric lymph node. Microscopic analysis showed that the lymph nodes in both dolphin species were surrounded by a connective tissue capsule comprising smooth muscle cells. The parenchyma of the mesenteric and bifurcational lymph nodes in bottlenose dolphin was divided into the peripherally situated cortex with the lymphatic nodules and diffuse lymphatic tissue, and the centrally situated medulla structured of the medullary cords separated by the medullary sinuses. These lymph nodes structurally correspond to the lymph nodes in the majority of terrestrial mammals. The mesenteric lymph node of striped dolphin also had a peripherally situated cortex and a centrally positioned medulla as the majority of terrestrial mammals. In the bifurcational lymph nodes of striped dolphin, there was a central dense lymphatic tissue with the lymphatic nodules and a peripheral less dense lymphatic tissue structured of the cell cords and sinuses. The bifurcational lymph node in striped dolphin resembled porcine lymph nodes and belonged to the inverse lymph nodes. PMID- 15782656 TI - Comparison of two-dimensional echocardiographic measurements of the left atrium in healthy dogs. AB - The aim of the study was to establish normal echocardiographic values of the left atrium just above the mitral annulus (LAama) in healthy dogs. In the first part of the study 20 dogs of various breeds were examined. The diameter of the left atrium just above the mitral annulus (LAama) was compared to the linear (left to right) dimension of the left atrium (LAr-l) as published earlier. There was a linear relationship between LAr-l and the body weight (LAr-l = 0.5061 BW (kg) + 22.206; R2 = 0.81), but the proportion of LAr-l/LAama was independent of the body weight (LAr-l/LAama = 0.0004 BW (kg) + 1.0833; R2 = 0.01). In the second part of the study the left atrial diameter just above the mitral annulus (LAama) was measured in 121 dogs of various breeds. There was a positive linear relationship (R2 = 0.697; p < 0.001) between body weight and LAama (LAama = 0.54 BW (kg) + 18.502+/-4.76), but there were no significant correlations between the age of animals and LAama (p = 0.45) as well as between the gender of animals and LAama (p = 0.78). Two-dimensional echocardiographic (2DE) determination of LAama as described in the present study can be recommended for use in those dogs where measurement of LAr-l encounters technical difficulties. In these cases LAr-l can be calculated from LAama using the formula LAr-l/LAama = 0.0004 BW (kg) + 1.0833, worked out in the first experiment. PMID- 15782655 TI - Identification of Borrelia burgdorferi genospecies inducing Lyme disease in dogs from Western Poland. AB - Canine Lyme borreliosis may be caused by three Borrelia burgdorferi sensu lato genospecies. The prevalence of infection by Borrelia species was determined by nested polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) with the enzyme Fsp4H I in the blood of dogs naturally infested by ticks in an endemic region of Poland. Blood samples were collected from 98 dogs of various breeds, delivered to the Veterinary Clinic in Szczecin (northwestern Poland) for various reasons. Nested PCR revealed the presence of DNA characteristic of only 1 genospecies, i.e. B. burgdorferi sensu stricto (s.s.), in all PCR-positive samples. Digestion of PCR products from a fragment of the fla gene amplified with primers FLA1 and FLA2 gave only one band pattern consistent with the pattern obtained from sequence analysis of the fla gene from a reference isolate of B. burgdorferi s.s. GeHo (X15660) from GenBank. PMID- 15782657 TI - Ochratoxin a content of urine samples of healthy humans in Hungary. AB - The ochratoxin A (OTA) content of urine samples from 88 healthy humans living at five settlements in three counties of Hungary was determined by immunoaffinity column cleanup and high-performance liquid chromatography (HPLC). OTA was detected in 61% of the samples in an average concentration of 0.013 ng/ml (range: 0.006-0.065 ng/ml). OTA concentrations measured in urine samples from men and women were not significantly different. The OTA concentration of samples from Heves county was significantly (t-test; p < 0.003) higher than that of samples from Hajdu-Bihar and Somogy counties. The regional differences in OTA concentration of urine samples indicate regional differences in the OTA exposure of the human population. Further studies are necessary to determine the cause of the regional differences in the OTA intake. The studies allow us to conclude that the OTA intake of the majority of the Hungarian population is low (< 1 ng/kg of body weight per day) but a certain part of the rural population may take up higher levels of OTA. PMID- 15782658 TI - Histopathological changes caused by the metacestodes of Neogryporhynchus cheilancristrotus (Wedl, 1855) in the gut of the gibel carp, Carassius gibelio. AB - Metacestodes of Neogryporhynchus cheilancristrotus (Wedl, 1855) were found in the gut of some gibel carp (Carassius gibelio) specimens from a Hungarian water reservoir. Location of metacestodes in the freshly opened gut was marked with disseminated, red-coloured, pinhead-sized nodules in the anterior part of the intestine. In histological sections, metacestodes were found in a hole inside the propria layer of the intestinal folds. The worms were in direct contact with the host tissue without being encapsulated as a result of host reaction. In some specimens with extruded rostellum the rostellar hooks were bored into the host tissue and suckers grabbed pieces of the surrounding connective tissue. Around the worms, congested capillaries and formation of macrophages were seen in the lysed connective tissue. PMID- 15782659 TI - Investigation of lectin activity in Theileria annulata piroplasms. AB - Adhesion to target cells is an essential step in the pathogenesis of many protozoal infections. Some protozoa have been reported to have a lectin activity involved in their attachment to the cell surface. The ligand-receptor interaction involved in Theileria annulata infection is unclear at present, in spite of the fact that some aspects of the process have been investigated. To this end, T. annulata piroplasms have been screened for lectin activity. Blood taken from infected cattle was first depleted of leukocytes and then subjected to ammonium chloride lysis in order to isolate the piroplasms. The piroplasms were homogenised and a crude membrane extract was prepared by centrifugation. To investigate lectin activity in piroplasm proteins, a simple screening procedure was employed for analysing piroplasm proteins binding to various lectin ligands. Numerous immobilised lectin ligands (L-fucose-sepharose, N-acetyl-neuraminic acid sepharose, N-acetyl-D-galactosamine-agarose, N-acetyl-D-glucosamine-agarose, D mannose-agarose, beta-D-glucose-agarose, alpha-methyl-D-mannoside-agarose) were incubated with T. annulata piroplasm crude membrane extract. The ligand-bound proteins were eluted and separated by a brief centrifugation and determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The present study suggests that a 32 kDa protein of piroplasm binds to D-galactosyl residues of the agarose matrix and that the binding is inhibited by galactose and not by the other monosaccharides tested. PMID- 15782660 TI - Evaluation of efficacy of entomopathogenic nematodes against larvae of Lucilia sericata (Meigen, 1826) (Diptera: Calliphoridae). AB - The blowfly Lucilia sericata (Meigen, 1826) (Diptera: Calliphoridae) is the primary agent of cutaneous myiasis of sheep in northern Europe, southern Africa, Australia and New Zealand. As the application of chemicals has several disadvantages, alternative control measures of traumatic myiasis of livestock must be developed. In this study, the use of entomopathogenic nematodes (EPNs) as potential biocontrol agents against second instar larvae of Lucilia sericata was considered. The following nematode species were tested: Heterorhabditis bacteriophora (IS 5, HHU 1, Hmol, HNC 1, HAZ 36, Hbrecon, HHU 2, HAZ 29, HHP 88, HHU 3, HHU 4 and HGua), Steinernema intermedia, NC513 strain of S. glaserii, S. anomali, S. riobrave, Steinernema sp. and 5 strains of S. feltiae (22, Vija Norway, HU 1, scp, and IS 6). None of the examined EPN species or strains showed larvicidal efficacy at 37 degrees C (no killing effect was observed in the case of the two heat-tolerant strains--H. bacteriophora and S.feltiae) against L. sericata larvae. At lower temperatures (20 degrees C and 25 degrees C) only strains of S. feltiae were found to be active. The overall odds ratios calculated for L. sericata maggots to contract S. feltiae nematode infection show significant (p < 0.05) effect only in the case of strains HU 1, 22 and IS 6. In the case of strains HU 1 and 22 parasitic forms of S. feltiae could be detected in the dead larvae of L. sericata. Strain IS 6 (and also Vija Norway at 20 degrees C) penetrated and killed fly larvae, but only adult forms of the nematode occurred in the cadavers. PMID- 15782661 TI - Comparative pathological studies on domestic geese (Anser anser domestica) and Muscovy ducks (Cairina moschata) experimentally infected with parvovirus strains of goose and Muscovy duck origin. AB - Parvovirus infection of Muscovy ducks caused by a genetically and antigenically distinct virus has been reported from Germany, France, Israel, Hungary, some Asian countries and the USA. The pathological changes include those of degenerative skeletal muscle myopathy and myocarditis, hepatitis, sciatic neuritis and polioencephalomyelitis. In the study presented here, day-old and 3 week-old goslings and Muscovy ducks were infected experimentally with three different parvovirus strains (isolates of D-216/4 from the classical form of Derzsy's disease, D-190/3 from the enteric form of Derzsy's disease, and strain FM from the parvovirus disease of Muscovy ducks). All three parvovirus strains caused severe disease in both day-old and 3-week-old Muscovy ducks but in the goslings only the two strains of goose origin (D-216/4 and D-190/3) caused disease with high (90-100%) mortality when infection was performed at day old. Strain FM (of Muscovy duck origin) did not cause any clinical signs or pathological lesions in the goslings. In the day-old goslings and Muscovy ducks the principal pathological lesions were severe enteritis with necrosis of the epithelial cells (enterocytes) of the mucous membrane and the crypts of Lieberkuhn, and the formation of intranuclear inclusion bodies. Other prominent lesions included hepatitis and atrophy (lymphocyte depletion) of the lymphoid organs (bursa of Fabricius, thymus, spleen). In goslings infected with the strain originating from the classical form of Derzsy's disease mild myocarditis was also detected. After infection at three weeks of age, growth retardation, feathering disorders, myocardial lesions (degeneration of cardiac muscle cells, lympho histiocytic infiltration) and hepatitis were the most prominent lesions in both geese and Muscovy ducks. In addition to the lesions observed in the geese, muscle fibre degeneration, mild sciatic neuritis and polioencephalomyelitis were also observed in the Muscovy ducks infected with any of the three parvovirus strains. PMID- 15782662 TI - The effect of nerve growth factor on nuclear progression of porcine oocytes during in vitro maturation and embryo development. AB - The present study examined the effect of nerve growth factor (NGF) on in vitro maturation (IVM), in vitro fertilisation (IVF) and subsequent embryonic development of porcine oocytes. Cumulus-oocyte complexes were cultured with or without 1.0 ng/ml NGF for 40 h. After IVF, they were cultured in vitro for 6 days. After 10 and 20 h of IVM, there was no difference in nuclear status between the NGF-treated and control oocytes. Significant differences were detected in nuclear progression of oocytes matured in the presence or absence of NGF at 30 h of culture. A higher proportion of NGF-treated oocytes were at M-II stage compared to the control. Nevertheless, at the end of the 40-h IVM period, there was no difference in the proportion of M-II stage oocytes between the NGF-treated and control groups. NGF in IVM medium did not influence the developmental competence of putative embryos. Most embryos remained at the 2- to 4-cell stage; however, a significant amount of embryos reached the morula stage both in the NGF and the control groups. These results suggest that NGF during IVM accelerates nuclear progression of porcine oocytes by enhancing the post-diakinetic events of meiosis. PMID- 15782663 TI - Vitrification of biopsied mouse embryos. AB - Solid surface vitrification (SSV) was compared with in-straw vitrification for cryopreservation of biopsied mouse embryos. Eight-cell stage embryos were zona drilled and one blastomere was removed. Developed morulae or blastocysts were vitrified in microdrop (35% EG + 5% PVP + 0.4 M trehalose) or in straw (7.0 M EG + 0.5 M sucrose). Following recovery, embryos were cultivated in vitro or transferred into recipients. Cryopreservation had an effect not only on the survival of biopsied embryos but also on their subsequent development in vitro. Cryosurvival of biopsied morulae vitrified in straw was significantly inferior to SSV. The post-warm development of biopsied and non-biopsied morulae was delayed on Day 3.5 and 4.5 in both vitrification groups. A delay in development was observed on Day 5.5 among vitrified non-biopsied blastocysts. The percentage of pups born from biopsied morulae or blastocysts following cryopreservation did not differ from that of the control. No significant differences could be detected between methods within and between embryonic stages in terms of birth rate. The birth rate of biopsied embryos vitrified in straw was significantly lower compared to the non-biopsied embryos. The novel cryopreservation protocol of SSV proved to be effective for cryopreservation of morula- and blastocyst-stage biopsied embryos. PMID- 15782664 TI - The molecular diagnosis of porcine viral diseases: a review. AB - The worldwide occurrence and re-occurrence of transboundary diseases like foot and-mouth disease or classical swine fever indicates that there is a high need for the development of powerful, robust and high-capacity new diagnostic methods, which are able to detect the causative agents before they could spread to large populations and cause tremendous losses. This article reports the experiences of a research group on the development of molecular methods for the improved diagnosis of a range of porcine viral diseases, including diseases on List A of the Office International des Epizooties (OIE). Nucleic acid hybridisation and various polymerase chain reaction (PCR) assays have been applied for routine diagnosis of a large range of viral diseases. During the last one-and-a-half decade more than 40 nested PCR assays have been developed to detect a variety of DNA and RNA viruses. False positive and negative results are avoided by the use of special tools, practices and internal controls of amplification (mimics). Recently, real-time PCR methods (TaqMan, molecular beacons, Primer-Probe Energy Transfer system) have been developed for the diagnosis of a wide range of diseases, such as foot-and-mouth disease, swine vesicular disease and vesicular stomatitis. Multiplex PCR packages have been developed for the simultaneous detection of eight important viruses of swine. By introducing nucleic acid extraction and pipetting robotics, together with the multi-channel real-time PCR machines, the diagnostic procedures have become rapid, robust and automated. In order to standardise the real-time PCR assays, the rules of OIE are considered. By following the five steps of OIE standardisation and validation, the new diagnostic procedures are nationally and internationally standardised and harmonised. The rapid, powerful and internationally standardised molecular diagnosis contributes to the reduction of losses caused by the transboundary viral diseases in swine populations. PMID- 15782665 TI - Cytopathogenicity markers in the genome of Hungarian cytopathic isolates of bovine viral diarrhoea virus. AB - Since genetic recombination is a major factor in the evolution of the cytopathogenic (cp) bovine viral diarrhoea virus (BVDV) biotypes, in this study the cytopathogenicity markers were investigated in the genomes of two cp BVDV strains recently isolated from mucosal disease (MD) cases in Hungary. In the genome of strain H4956, a Jiv-like insertion was found similar to those described in reference strain NADL and in other BVDV 1, BVDV 2 and border disease virus (BDV) strains. The 133 amino acid Jiv-like sequence is inserted at nucleotide position 4984 (amino acid position 1533), 9 nucleotides upstream of that of strain NADL. The insertion showed 96% amino acid sequence identity with the cellular Jiv protein. In the genome of cp BVDV strain H115/PCR, an ubiquitin containing duplication was found. The duplicated sequence started at nucleotide position 7978 (amino acid 2531) in the NS4B gene. The duplication contained a complete ubiquitin monomer of 76 amino acids and the complete NS3 gene starting at nucleotide position 5153 (amino acid 1589), which corresponds to the first N terminal amino acid of NS3. The duplication was located further downstream of the known ubiquitin-containing genomic regions of cp BVDV strains, and it consisted of the shortest inserted nucleotide sequence. The insertions and duplication of strains H4956 and H115/PCR further confirmed that recombinations occurring at positions A and B are the most common mechanisms leading to the development of BVDV cytopathogenicity. PMID- 15782666 TI - Detection of respiratory and enteric shedding of bovine coronaviruses in cattle in Northwestern Turkey. AB - Bovine coronavirus (BCoV) is an important cause of diarrhoea in calves, winter dysentery in adult cattle and respiratory tract disease in feedlot cattle. Serum, faecal and nasal swab samples were collected from a total of 96 cattle with clinical signs in 29 barns of 23 villages in Northwestern Turkey. The cattle were subdivided into 3 distinct age groups (0-30 days old, 4-12 months old and 2-7 years old). An indirect antigen-capture ELISA and an antibody-detection ELISA as well as geometric mean BCoV antibody titres were used to detect BoCV shed in the faeces and in the nasal secretions, respectively. Relationships between BCoV shedding and age group, seroconversion and clinical signs in cattle were also analysed. The rate of faecal shedding of BoCV was 37.1% (13/35) in 0-30 days old calves, 25.6% (10/39) in 4-12 months old feedlot cattle and 18.2% (4/22) in 2-7 years old cows. The overall rate of BCoV faecal shedding was 28.1% (27/96) in the cattle examined. Only one animal in the 4-12 months old age group was found to shed BoCV nasally. The analysis showed that there was a significant difference (P < 0.0001) with respect to faecal shedding between the clinical signs and the age groups. BCoV antibody titre in 50% of all cattle was < or =100 as detected by ELISA while 27.1% of the cattle had high titres ranging between 1,600 and 25,600. The seroconversion rate was 7.3% (7/96) in animals shedding BoCV in the faeces and 42.7% (41/96) in cattle negative for faecal shedding as detected by ELISA, and 20.8% of cattle with no seroconversion shed BCoV in the faeces. There was no statistically significant association between seroconversion and nasal or faecal BCoV shedding. These findings confirm the presence of BCoV infections in Turkey. Further studies are needed to isolate BCoV strains in Turkey and to investigate their antigenic and genetic properties. PMID- 15782667 TI - The replication of canine herpesvirus (CHV) induces apoptosis in canine kidney cell line: short communication. AB - The alphaherpesvirus canine herpesvirus (CHV) was tested in order to determine whether or not it has apoptotic potential. We have demonstrated that lytic replication of CHV resulted in induction of apoptosis. This phenomenon was confirmed using different techniques including in situ TUNEL assay and DNA laddering. The apoptotic activity of CHV might influence the pathobiology of this virus. PMID- 15782668 TI - Latent autoimmune diabetes of adulthood. Unique features that distinguish it from types 1 and 2. AB - A number of patients with poor glycemic control receive the diagnosis of type 2 diabetes despite the fact that they do not exhibit some of the traditional characteristics of the disease, such as obesity. A more accurate diagnosis for many of these patients is latent autoimmune diabetes of adulthood (LADA). In this article, Dr Nabhan and coauthors describe features that LADA has in common with type 1 and type 2 diabetes, as well as those that distinguish LADA from these more widely recognized forms of diabetes. The authors also describe the pathogenesis of the disease, potential complications, and treatment options. PMID- 15782669 TI - Addressing the growing menace of COPD. PMID- 15782670 TI - Is it asthma or COPD? The answer determines proper therapy for chronic airflow obstruction. AB - Evaluation and diagnosis of obstructive lung disease call for careful history taking to distinguish asthma from chronic obstructive pulmonary disease (COPD). A history of atopy and intermittent, reactive symptoms points toward a diagnosis of asthma, whereas smoking and advanced age are more indicative of COPD. Clinical features such as cough and wheezing, especially in light of patient history, can be suggestive but are not definitive. Spirometry, usually performed in the primary care setting, is the key diagnostic tool that distinguishes patients with asthma from patients with COPD. In this article, the authors outline an approach to differential diagnosis of asthma and COPD that should result in better evaluation, therapy, and quality of life for these patients. PMID- 15782671 TI - New treatment strategies for COPD. Pairing the new with the tried and true. AB - Knowledge about COPD has increased substantially in recent years. Smoking cessation campaigns have significantly decreased smoking prevalence in the United States, and similar efforts in the rest of the world will likely have the same impact. The consequence should be a drop in incidence of COPD in the years to come. The use of LTOT for hypoxemic patients has resulted in increased survival, and expanded drug therapy options have effectively improved dyspnea and quality of life. Recent studies have documented the benefits of pulmonary rehabilitation. In addition, noninvasive mechanical ventilation offers new alternatives for patients with acute or chronic failure. Furthermore, the revival of surgery for emphysema may serve as an alternative to lung transplantation for patients with severe COPD who remain symptomatic despite maximal medical therapy. With all of these options, a nihilistic attitude toward management of COPD is not justified. PMID- 15782672 TI - Posterior vitreous detachment. How to approach sudden-onset floaters and flashing lights. AB - Symptomatic PVD is a common age-related problem that in some patients is associated with retinal tear and retinal detachment. Thorough ocular examination of patients with unilateral floaters and photopsias is needed to identify those at greatest risk for vision loss. Patients without retinal pathologic findings on initial examination need to be given careful instructions on what to expect and do should symptoms of retinal tear or detachment occur. PMID- 15782673 TI - Seborrheic dermatitis. The link between facial rash and neurologic disease. PMID- 15782674 TI - Rapid HIV testing. Wait time reduced from days to minutes. AB - Recent breakthroughs in technology have produced tests for HIV antibody that are highly accurate and easy to use and can give a preliminary result in 20 minutes or less. These rapid HIV tests will be used increasingly in labor and delivery wards, emergency departments, urgent care centers, and the primary care office. They have unique applications for healthcare worker exposures, military operations, public health venues, and developing countries. In this article, Drs Keenan, Keenan, and Branson discuss the advantages and limitations of rapid HIV testing in various settings. PMID- 15782675 TI - The relationship of motivation and flow experience to academic procrastination in university students. AB - In this article, the author examined the relationships of motivation and flow experience to academic procrastination in 262 Korean undergraduate students who completed a questionnaire on procrastination, flow, and motivation. The results indicated that high procrastination was associated with lack of self-determined motivation and low incidence of flow state. The results also indicated that, although amotivation and intrinsic motivation showed significant unique effects on procrastination, they did not contribute significantly to the variance in procrastination when the effects caused by flow experiences were considered. The author discusses implications for practice and gives suggestions for further research. PMID- 15782676 TI - Moderation and mediation effects of coping by Chinese American and European American adolescents. AB - The authors asked 55 second-generation Chinese American adolescents (M age = 16.8 years) and 58 European American adolescents (M age = 17.0 years) to complete self report measures of stress levels, use of coping strategies, psychological outcomes, and grades. Chinese American adolescents reported higher levels of everyday life event stress (e.g., stress from schoolwork), more depression, and higher grade point averages. Problem-focused and avoidance-coping behaviors moderated the effect of stress on negative adjustment for Chinese American youth but not for European American youth. Path analysis showed that avoidance coping behavior partially mediated the stress-to-negative adjustment relationship for Chinese American youth but not for European American youth. Stress was associated with lower grades in Chinese American youth, but the authors found no association for European American youth. PMID- 15782677 TI - The development of interindividual sharing of knowledge and beliefs in 5- to 9 year-old children. AB - The authors studied the evolution of interindividual intentionality in children and showed that the sharing of knowledge and beliefs requires more complex operations than those involved in usual false-belief tasks. The authors conducted 3 experiments on 380 children (aged 5 years, 0 months to 9 years, 6 months). They assessed the children's ability to attribute to a character an intended action that was compatible with the belief held by another character interacting with that character. The observed lag in performances is explained both in terms of information processing and at the Piagetian concrete operational level. PMID- 15782678 TI - Developmental stability and change in self-regulation from childhood to adolescence. AB - The authors examined the developmental course of self-regulation in a cohort of children from the National Longitudinal Survey of Youth. The longitudinal sample included 646 children (48% girls; 52% boys; 36.2% Black, 23.4% Hispanic, 40.4% White) who were 4 to 5 years old in 1986 and who were followed up at ages 8 to 9 and ages 12 to 13. Levels of self-regulation (assessed with 12 maternal-report items that measured regulation of affect, behavior, attention) increased from early childhood (when sample children were 4 or 5 years old) to middle childhood (ages 8 or 9), but not from middle childhood to early adolescence (ages 12 or 13). Girls exhibited significantly higher levels of self-regulation than did boys at all 3 time points. Individual differences in self-regulation were fairly stable across the 8-year span (rs = .47 to .50). Comparisons of 1-, 2-, and 3 factor models suggested that the different aspects of self-regulation are highly interrelated, and support adoption of a single-factor model for both genders. The authors discuss implications of these findings for theory and intervention. PMID- 15782679 TI - The Draw-A-Person Test: an indicator of children's cognitive and socioemotional adaptation? AB - The authors examined aspects of reliability and validity of the Goodenough-Harris Draw-A-Person Test (DAP; D. B. Harris, 1963). The participants were 115 seven- to nine-year-old students attending regular or special education schools. Three judges, with a modest degree of training similar to that found among practicing clinicians, rated the students' human figure drawings on developmental and personality variables. The authors found that counting details and determining developmental level in the DAP test could be carried out reliably by judges with limited experience. However, the reliability of judgments of children's social and emotional development and personality was insufficient. Older students and students attending regular schools received significantly higher scores than did younger students or students attending special education schools. The authors found that the success of the DAP test as an indicator of cognitive level, socioemotional development, and personality is limited when global judgments are used. The authors concluded that more specific, reliable, valid, and useful scoring systems are needed for the DAP test. PMID- 15782680 TI - Social skill in self-assertive strategies of toddlers with depressed and nondepressed mothers. AB - The authors examined the effects of maternal depression on the skillfulness of toddlers' self-assertive strategies in interactions with their mother and with a female examiner. The participants were 110 mothers and their 26-month-old toddlers. Of these mothers, 57 had experienced an episode of clinical depression sometime since their child's birth, and 53 had had no history of depression. Toddlers exposed to maternal depression demonstrated significantly less social skill in their self-assertive strategies and more defiance when interacting with their mothers than did toddlers who were never exposed to maternal depression. The chronicity and severity of toddlers' exposure to maternal depression did not account for more pronounced differences in toddlers' skill in self-assertion toward mothers; however, toddlers exposed to more chronic courses of depression demonstrated less skill in their self-assertion toward the examiner. Toddlers who were exposed to maternal depression with a comorbid anxiety disorder did not exhibit less skill in their self-assertion toward mothers than did toddlers in either the depression-only or nondepressed groups. These findings suggest that exposure to maternal depression may interfere with toddlers' development of socially competent self-assertion strategies and may pose risks for future problems in the mother-toddler relationship. PMID- 15782681 TI - The effects of stating problems in bilingual students' first and second languages on solving mathematical word problems. AB - Researchers have suggested that among bilinguals, solving word problems in mathematics is influenced by linguistic factors (K. Durkin & B. Shire, 1991; L. Verschaffel, B. Greer, & E. De Corte, 2000). Others have suggested that students exhibit a strong tendency to exclude real-world constraints in solving mathematics word problems (L. Verschaffel, E. De Corte, & S. Lasure, 1994). In the present study, the authors explored the effects of stating word problems in either Filipino or English on how Filipino-English bilingual students solved word problems in which the solution required the application of real-world knowledge. The authors asked bilingual students to solve word problems in either their first or second language. For some of the word problems, real-life constraints prevented straightforward application of mathematical procedures. The authors analyzed the students' solutions to determine whether the language of the word problems affected the tendency to apply real-life constraints in the solution. Results showed that the bilingual students (a) rarely considered real-life constraints in their solutions, (b) were more successful in understanding and solving word problems that were stated in their first language, and (c) were more likely to experience failure in finding a solution to problems stated in their second language. The results are discussed in terms of the relationship between linguistic and mathematical problem-solving processes among bilinguals. PMID- 15782683 TI - [The practice guideline 'Depressive disorder' (first revision) from the Dutch College of General Practitioners; a response from the perspective of general practice]. AB - The revised Dutch College of General Practitioners' practice guideline 'Depressive disorder' no longer distinguishes between mild and severe depression because the management in both cases is identical. This guideline focuses much attention on diagnosis, support and information, all of this in dialogue with the patient. The choice between the antidepressants, a tricyclic antidepressant or a specific serotonin reuptake inhibitor is based on the contraindications of these drugs. PMID- 15782682 TI - [Timing and choice of intervention in necrotising pancreatitis]. AB - Three patients, men aged 49, 62 and 33 years, were admitted with acute abdominal symptoms due to necrotising pancreatitis. They underwent multiple interventions during a hospital stay of several months, but ultimately recovered completely. In case of infected (peri-)pancreatic necrosis, intervention is required. Good clinical judgement in the differentiation between the septic inflammatory response syndrome, sepsis and infected necrosis as the cause of the clinical condition is important. Because of the different intervention strategies, treatment by a team comprising a radiologist, gastroenterologist, intensive care specialist and gastrointestinal surgeon is required. Randomised studies on intervention in infected pancreatic necrosis are lacking. In 2002, to improve the treatment of patients with acute (necrotising) pancreatitis via a combination of research, consultation and centralisation, the Dutch Acute Pancreatitis Study Group was formed. PMID- 15782684 TI - [The practice guideline 'Depressive disorder' (first revision) from the Dutch College of General Practitioners; a response from the perspective of psychiatry]. AB - The first revision of the Dutch guidelines for the treatment of depressive disorders by general practitioners adapted the international psychiatric nomenclature for the diagnosis of depressive disorders. This enhances collaboration between general practitioners and psychiatrists. The use of selective serotonin reuptake inhibitors in this guideline is an advantage compared to the earlier version. The distinction between the indication for cognitive therapy and for antidepressant treatment is not very clear in this guideline. Overall the guideline represents a scientific and conservative stance in the treatment of depressed patients, which is commendable. The use of target plasma level concentrations with tricyclic antidepressants, better guidelines for the use of antidepressants or cognitive therapy, and the use of severity rating scales during treatment could further improve the next version of this guideline. PMID- 15782685 TI - [Skin cancer and other skin disorders in patients following solid organ transplantation]. AB - Solid organ transplant patients have an increased risk of cutaneous squamous cell carcinomas compared to the immunocompetent population, and often develop multiple and sometimes aggressive tumours. There are few published studies or reviews, which provide guidance to the clinician in the management of these patients. In the prevention of skin cancer in organ transplant patients, patient education about the harmful effects of ultraviolet radiation, sun protection, and the early recognition of (pre)malignant skin lesions should be emphasised. Furthermore, close follow-up by a dermatologist and treatment of (pre)malignant lesions in an early stage are necessary. Chemoprevention of skin cancer can be achieved through systemic retinoids. Reduction of the dose of immunosuppressive agents can be considered. Excision is the first treatment of choice for squamous cell and basal cell carcinomas. In selected rumours curettage and electrodessication can be performed. PMID- 15782686 TI - [Human papillomavirus in the aetiology of skin cancer]. AB - At present, human papillomavirus (HPV) infection is chiefly known for its causal relationship with cervical cancer. Apart from genital types, the papillomavirus family consists of numerous human cutaneous types. The majority belongs to the so called epidermodysplasia-verruciformis(EV)-HPV types that are potentially involved in skin cancer development. Non-melanoma skin cancers, especially cutaneous squamous cell carcinoma contain HPV DNA (30-60%). In immune-suppressed organ transplant recipients this percentage increases up to 90. Recent epidemiological studies show a statistically significant association between EV HPV infection and squamous cell carcinoma. In addition recent experimental studies show specific EV-HPV types have a potential to transform cells that is comparable to high-risk genital HPV types. These data indicate that cutaneous HPV infections and squamous cell carcinoma development are associated. PMID- 15782687 TI - [Summary of the practice guideline 'Depressive disorder' (first revision) from the Dutch College of General Practitioners]. AB - In the revised practice guideline for the diagnosis and treatment of patients with a depressive disorder in general practice the distinction between mild and severe depression has been removed. By paying attention to a number of risk factors, the general practitioner can detect patients with a hidden depressive disorder. The general practitioner's strategy is aimed at reducing symptoms and restoring the ability to function normally. Factors that define the strategy are in particular patient's degree of suffering and dysfunctioning, and the patient's preferences and demands. In primary care tricyclic antidepressants and specific serotonin re-uptake inhibitors are the drugs of first choice: both are equally effective; the nature of the side effects differs. PMID- 15782688 TI - [Diagnostic image (230). A man with bilateral shoulder dislocation]. AB - A 28-year-old man suffered acute primary posterior dislocation of the left shoulder and anterior dislocation of the right one due to a motor accident. PMID- 15782689 TI - [Direct reporting of side effects by the patient: favourable experience in the first year]. AB - OBJECTIVE: To determine the results of establishing a station at which patients can report the side effects of drugs. DESIGN: Descriptive. METHOD: Since 1 April 2003, patients may submit reports of possible adverse drug reactions directly to the Netherlands Pharmacovigilance Centre Lareb. The reports submitted during the period from 1 April 2003 to 31 March 2004 were analysed and compared with the reports submitted by doctors and pharmacists. RESULTS: In the first year, 276 reports were submitted by patients and 3131 by doctors and pharmacists. The reports from patients usually contained sufficient medical information and more frequently referred to serious adverse reactions than reports by health professionals. The reports from patients relatively often concerned psychotherapeutic agents, notably antidepressants. CONCLUSION: Based on the positive results during the first year, the Netherlands Pharmacovigilance Centre Lareb has decided to continue the reporting station for patients. Reports submitted by patients are currently part of the core responsibility of Lareb: the detection of signals of new adverse drug reactions. PMID- 15782690 TI - [Three men with breast cancer]. AB - In three men, aged 81, 66 and 58 years, breast cancer was diagnosed. All three were treated by modified radical mastectomy and axillary-node dissection. They received tamoxifen and the first and the third patient also received radiotherapy. Presentation of these patients is usually delayed, because of the rarity of and unfamiliarity with the disease. Consequently, the disease is often in a more advanced stage at presentation and overall mortality is higher in comparison with women with breast cancer. However, survival of male patients is similar to survival of female patients when matched for age and stage. This stresses the importance of a timely diagnosis. As in women, of the known risk factors for male breast cancer, a positive family history is one of the most important ones. Modified radical mastectomy, combined with sentinel-node biopsy by experienced teams, is the standard treatment. Criteria for adjuvant systemic treatment are identical for men and women, although hormonal therapy (tamoxifen) has a more prominent place in the adjuvant setting because of the high percentage of positive hormone receptors in men. PMID- 15782691 TI - [Erection due to 'high flow' priapism in a 5-year old: an emergency]. AB - A healthy 5-year-old child was brought to the general practitioner with priapism that had been observed for a few days, without any other symptoms. Angiography revealed an arteriovenous fistula originating from the left A. dorsalis penis, which was successfully embolised. The diagnosis was 'high flow' priapism. A possible cause in the anamnesis was a fall from a swing two weeks before the embolisation. The doctor's delay was caused by unfamiliarity with this rare disorder; due to the risk of impotence (impotentia coeundi), it is an emergency situation. PMID- 15782693 TI - [The practice guideline: 'Treatment of tobacco dependence']. PMID- 15782692 TI - [Aggression towards paediatricians and trainee paediatricians in the Netherlands]. AB - OBJECTIVE: To determine the extent of aggression from the patient or his family to which paediatricians and trainee paediatricians in the Netherlands are exposed. DESIGN: Cross-sectional survey. METHOD: Data were collected using anonymous questionnaires distributed to doctors working in the field ofpaediatrics in the Netherlands during the annual congress of the Paediatric Association of the Netherlands in 2002. RESULTS: A total of 395 usable questionnaires were returned. 78% of the respondents reported that they had, at some time, been confronted with aggression. Verbal aggression was the most common form (75% of the respondents) and was experienced mainly by doctors with little work experience (63% in the preceding year). 20% of the respondents were aware of the existence of training programmes on dealing with aggression in their hospitals, whereas 47% said that they needed such training. CONCLUSION: Aggression in connection with paediatric care was reported by 78% of the respondents, especially verbal aggression directed at doctors with little work experience. Training on how to cope with aggression might be a useful addition to the paediatric curriculum. PMID- 15782694 TI - [Practice guideline 'Influenza prevention in nursing homes and care homes', issued by the Dutch Society of Nursing Home Specialists]. PMID- 15782695 TI - [How should age be documented?]. PMID- 15782696 TI - [Crying upon eating: the crocodile-tears syndrome]. PMID- 15782697 TI - [Job-related accidents with chemical substances among nursing workers]. AB - This study aimed at analyzing job-related accidents with chemical substances among nursing workers. The study was performed at the University of Sao Paulo (USP) Hospital. In order to collect data, a form was used to survey accidents occurred between 1992 and 2001. Data analysis showed that accidents notified at the USP Hospital are presented in such a way not to allow an in-depth questioning of the job-related exposition to chemical substances, given the fact that the hospital environment hosts hundreds of chemical substances that can potentially cause accidents. Although notifications are few, they are rich in terms of content for analysis, which contributes to subsidize valuable and viable prevention proposals. PMID- 15782698 TI - [How the family acts regarding the health-disease process of a family member with breast cancer]. AB - Qualitative study performed with ten relatives of cancer patients in two clinics located in Feira de Santana-BA, aiming at analyzing how the family thinks, feels, and acts concerning the disease. Data was collected by means of a semi-structured interview and analyzed by dialectic hermeneutics. Results showed that most interviewees associated cancer and death, and the first reaction concerning the disease was unexpected, with a feeling that "the world has fallen". With time, there appeared hope, solidarity and faith to face the disease; however, they regarded emotional support as critical for the treatment. Family experience regarding breast cancer was seen as a "life change" in individuals, in emotional, religious and biological terms. Such changes showed the need for support groups to help patients and their relatives to face the disease. PMID- 15782699 TI - [Competency for teaching and learning nursing diagnosis]. AB - This is a qualitative, exploratory, descriptive and interpretative study. The objective of the study is to relate the conceptions that professors and students have over competencies with the teaching-learning process of nursing diagnosis. The study was carried out at the Nursing School of the Federal University of Rio Grande do Sul. The information was collected in two stages. The first one through eight semi-structured individual interviews and the second one through two focus groups. One of them was formed by students and the other group was formed by professors. Content analysis was used as analysis methodology. As a result three categories were found: relation with knowledge, teaching-learning strategies and institutional context of the course. PMID- 15782700 TI - [Knowledge produced about assistance to pregnant adolescents]. AB - Aiming at getting to know the work of nurses concerning pregnant adolescents, we have opted to develop a study to identify the characteristics of scientific production made by nurses about adolescence pregnancy by means of national and international indexed articles, between 1996 and 2000. The bibliographic survey resulted in a sample of 27 indexed articles. The participation of nurses in assisting pregnant adolescents is highlighted by taking part in adolescence pregnancy prevention programs, by assisting in the gravidic-puerperal period and by providing social support. As possibilities of participation for nurses, we have identified the important role of sexual education and adolescence pregnancy prevention programs, prenatal and post-delivery assistance programs. Also, we have emphasized the need to organize the nurse's work to facilitate access to contraceptive methods and to prevent sexually transmissible diseases. PMID- 15782701 TI - [Nursing care in an adverse situation]. AB - It is a final research report in which we use the quanti-qualitative approach by means of an exploratory study aiming at the surveillance usually carried out in control and prevention actions by a committee of hospital infection control and its interaction with the nursing team of surgery sectors. In the first stage, there was the survey of opinions issued for this committee, surveillance of dialyses procedures, teaching and extra-hospital activities, participation in meetings, accidents with biologic material and momentaneous prevalence. In the second stage, risk factors that might contribute to this installation with the participation of 22 nursing assistants and nine nurses. We have concluded that general conditions in which the customer is admitted in this scenario contribute to this occurrence and involve mostly, besides the physical space as a therapeutic environment, the improvisation of materials and that the nursing care should be executed in a participatory fashion. PMID- 15782702 TI - [Management of new technologies in a surgery center by nurses in hospitals located in Feira de Santana - BA]. AB - It is a quantitative, exploratory, descriptive study about the level of adequacy of the command of new technologies (NADNT) by nurses in the management of a surgery center in hospitals located in Feira de Santana-BA, Brazil, in 2002. It aimed at: assessing the NADNT of nurses in the management of a surgery center; identifying and describing the factors that interfere with this level; and proposing actions for the development of this level in the management of surgery centers. Sample made up of 33 nurses with experience in surgery centers. A questionnaire was used for data collection. It has been concluded that the NADNT was unsatisfactory for 54.5% of the study subjects. It has been statistically proved that the capacity of the hospital and the working load are factors that interfere with the command of new technologies in surgery centers. PMID- 15782703 TI - [Aged with sarcopenia: an approach of nursing care]. AB - In the grow old process organic and functional damages happening, wich rhythm and intensity change from person to person and that finish to reduce the ability from each person has to adapt the environment, making more visible and vulnerable to illness process, that are the perturbation produced in the body by the illness taking the person to a natural physical decline. Pain and disorders in the muscle skeletal system are the most frequent complain in the old age and the second cause of inability in this group. This fact is comprehensible now that many diseases of the muscle skeletal system have more incidence with the advance of age, beside others that are almost exclusive of these population and are chronic illness, not deadly with cumulative frequence ones predisposing the others. Is in this context that is situated the responsability of the attention and care of the older with sarcopenia, having to the nursing like member of a interdisciplinar group, make efforts to act in the prevention, guidance, supervision and management of the assistance. PMID- 15782704 TI - [Models of assisted deliveries and women's participation]. AB - The study aimed at analyzing four proposals of obstetric assistance that co existed in the city of Rio de Janeiro from the early Thirties to the Fifties, as well as discussing the inclusion of graduated and non-graduated accoucheuses, and nurses. It is of a historical-social nature, making use of the written document as a primary source. The analysis is based Pierre Bourdieu's main concepts. With this research, it was possible to make evident the domiciliary obstetric service of the sanitarist model, the philanthropic proposal of society's ladies, the individualized hospital-based assistance, and the proposal of integrated obstetric assistance. The study also allowed the understanding of the exclusion of non-graduated women from the Brazilian obstetric field. PMID- 15782705 TI - [For a history of the present of playing and health practices]. AB - The purpose of this study is to reflect on the need to perform a historical discursive approach to contextualize and analyze in a critical way hospital and nursing discourses and practices related to spaces to play in pediatric infirmaries at hospitals. The Statute of the Children and Adolescents, made into a Federal Law in 1990, is a document that guides a new vision concerning the field of childhood and youth. Among the consequences of the ECA, there is the establishment of a different mode for the adult to relate with children, and that the effects on healthcare should assume and effect changes and transformations in the hospital scenario. We have tried to show the need to problematize and contextualize how our hospital scenario is determined by discourses that operate silently. In order to resist to the effects of such devices, health and nursing practices should be rethought and rectified in their respective disciplinary devices. PMID- 15782706 TI - [The nurse's participation in implementing the Family Health Program (PSF0 in Belo Horizonte]. AB - This study analyzes the nurse's participation in implementing the Family Health Program (PSF) in Belo Horizonte and aims at understanding their inclusion and expectations as an agent in the organization of health services and in overcoming problems found in the implementation of the program in a large city. A qualitative approach is used based on dialectic historic materialism. Eleven nurses participating in the PSF were the subjects of the research. Data was collected by means of a semi-structured interview and submitted to a discourse analysis treatment. The results demonstrate that the nurse has been involved with family health and playing a significant role ever since the implementation of the Community Health Agents Program (PACS) and that their inclusion in the PSF is favored both by their generalist background and their experience in planning, executing and evaluating health actions. Factors related to the nurse's work conditions, such as inadequacy of the environment, differentiated salary and treatment in relation to the doctor in team activities and the overload of tasks and responsibilities stand out as difficulty factors in implementing the PSF in Belo Horizonte. PMID- 15782707 TI - [The nurse's perceptions on administrative actions in their work process]. AB - A qualitative, exploratory study carried out from March to December 2002 in a university hospital based in the far south of the country, which aimed at identifying the nurses' perception on the administrative actions exercised in their daily work. Thematic categories emerged from data survey and content analysis in the interviews carried out with 10 nurses. We emphasize Category I: Administrative Actions as a Management Instrument; and Category II: Administrative Actions as Direct/indirect Care. In the first one, administrative actions are perceived as planning, coordination, leadership, control, information retention, and organization; the second one depicts administrative actions as an integral part of nursing care. It has been observed that many administrative actions exercised by nurses are inherent to their role, in keeping with the Nurse's Professional Exercise Code, and they are critical for care. PMID- 15782708 TI - [Profile of nursing workers with RSI/WRMD diagnosis in Salvador-Bahia in 1998 2002]. AB - This quantitative study aimed at drawing the profile of nursing workers with a confirmed RSI/WRMD diagnosis, seen at the Center of Workers' Health Studies (Cesat) in Salvador, Bahia, from 1998 to 2002. Data collection was performed based on medical reports and, complementarily, guidelines with items of interest for this investigation. Out of the medical reports, only those of nursing workers were selected. There was the identification of 79 workers--four nurses, 74 assistants, and one technician. There was a prevalence of women aged between 30 and 49, with high school and a monthly income from three to six minimum wages. Out of the total numbers, at the time of data collection, 77.22% were employed, 15.19% were unemployed, and 7.5% were retired. Among the injuries found, carpal tunnel syndrome and cervicalgia prevailed. PMID- 15782709 TI - [The critical-reflexive teaching-learning process]. AB - This study aimed at showing how the learning process is being built in the FAMEMA nursing course and identifying the different aspects of this process perceived by students based on the logics of the political-pedagogical project. The subjects of the research were fourth-grade students in the course. Research techniques used were focus groups and a semi-structured interview. Empirical material was organized according to the collective subject discourse technique, followed by the application of content analysis in the thematic modality. In the analysis of empiric material, it was highlighted that learning occurs in a process with successive approaches, throughout life, being graduation the beginning of a process. They experienced things in which they shared knowledge, conflicts, group work and teamwork. PMID- 15782710 TI - [Reflecting on the institutionalized elderly]. AB - Qualitative and descriptive research that aimed at identifying how the elderly family relationship has been taking place by identifying the reasons that led family members to opt for the institutionalization of their elderly, obtaining also the view of the family about home caring. Data collection was performed by using an open questioning instrument. Data collection was performed from August 2002 to July 2003. The subjects of the research were relatives of elderly people living at a home in a town in the northwestern region of Rio Grande do Sul. Through content analysis, it was seen that the greatest difficulty found by the family in spending time with their elderly relative is the "lack of time to care" for him, because most of them require full-time caring. PMID- 15782711 TI - [Interpersonal relationship of nursing interns in the hospital context]. AB - The object of this study is the interpersonal relationship of nursing interns of FENF/UERJ with customers, teachers, and the multidisciplinary team of a university hospital. The introduction of the concept of teaching-learning based on the critical education theory promoted a narrower relationship between teachers, students and other parties involved in working at a hospital. By using qualitative descriptive research whose subjects were 26 nursing interns of the 9th period in the 2nd semester of 2002, we aimed at validating observations about changes in the quality of students' interpersonal attitude. We concluded that the problematization methodology provides positive changes in the interpersonal attitude of interns by instrumentalizing them for the professional life based on respect and understanding of the particularities of subjects involved in the work process. PMID- 15782712 TI - [Therapeutic touch for the healing of skin injuries in guinea pigs]. AB - This study verifies the action of therapeutic touch--a complementary practice- over the process of healing of injuries caused in the skin of guinea pigs. It is a quantitative study of the experimental kind, and there was a comparison of healing speed among 10 guinea pigs that received water energized with such technique (group B) and 10 guinea pigs that received water with no treatment (group A). The project was executed in the experimental laboratory of the Nursing School at the University of Sao Paulo, and the length of the bruises was measured five times, at every four days. Among the finds, it was observed that, after 20 days, 100% of the guinea pigs in the group B had their bruises fully healed, which did not happen to 4 guinea pigs (40%) in group A. PMID- 15782713 TI - [Work and alcoholism: study with civil servants]. AB - This investigation was carried out with civil servants, and it aimed at analyzing the influence of labor in the production/ reproduction and transformation of alcoholic behavior. Methodologically, this study is characterized as exploratory and qualitative. Data collection was done by means of documental analysis and participatory observation. Interviews were performed if needed to complement the information. The identification of nexuses of production and reproduction of alcoholic behavior in an institutional context provided proposals for changing it, as well as showed the need to carry out an integrated job among the services that provide care for alcoholics by seeking other investments for gradually changing this behavior. PMID- 15782714 TI - [The path of domestic violence in the town of Ribeirao Preto]. AB - This study aims at learning and characterizing the path of violence cases referred from the Center of Reference of Children and Adolescents (CRCA) to the Center of Attention to Victimized Children and Adolescents (CACAV) between 1997 and 2001. The investigation performed was exploratory and descriptive. 46.8% of the cases referred to the CACAV (n = 731) involved women. The research also showed that there was prevalence in the cases of negligence, with 35.7%, differently from what is found in literature. We have concluded that there is a need to standardize terms and create an instrument of record-keeping that may allow the collection of accurate information on violence against children and adolescents. PMID- 15782715 TI - [Domestic and institutional violence in health services: women's experiences]. AB - Developed in state capitals Recife, Salvador and Aracaju, in three state maternity hospitals, this quantitative study aimed at identifying the manifestations of domestic and institutional violence suffered by women assisted in subprograms of the Program of Integral Assistance to the Woman's Health. Data collection was performed by using a semi-structured form. The sample featured young women, housewives, and those that are economically dependant on their significant others. Most women experience matrimonial violence, and financial dependency is the most often mentioned reason to bear it. The lack of vacancies characterized institutional violence due to omission. Research showed that experiencing such violence brings health problems to women, or aggravates them. PMID- 15782716 TI - [Nursing knowledge]. AB - It is a review of literature on nursing knowledge that has its main focus on the classification of the four patterns of knowledge identified and proposed by Carper: empirical, ethical, esthetic, and personal. Research material was made up of articles published in national and international periodicals in the two last decades of the 20th century. The analysis points towards the extension of patterns of knowledge, with the addition of three others, besides those initially proposed: sociopolitical, historical, and the unknown. PMID- 15782717 TI - [The issue of death and dying]. AB - This study brings about a reflection on death and dying, commented as if they had the same meaning. Nowadays, one talks about death as seldom as possible and, by its negation, there appeared the negation of diseases, thus causing the diseased people to become totally dependant on health teams. Technological and scientific advances led to postponing death, and it is possible to temporarily extend what humans decided to call life. This has become the great piece of criticism by scholars that seek to lead death again to its place by trying to bring it again into the center of reflections, analyses and discussions within the most diverse areas. PMID- 15782718 TI - [Challenges in the implementation of a curriculum by competencies]. AB - This article aims at describing and analyzing the experience of teachers of a state school located in Pernambuco in the implementation of the curriculum by competencies in the technical nursing course. National guidelines for technical level professional education were used as a theoretical referential. The technical nursing course was organized in four non-terminal modules, with a growing complexity in contents and specific competencies. Among the greatest difficulties found during curricular re-structuring, there are the teachers' and students' resistance concerning the adoption of new pedagogical practices and of a methodology to assess the constructed competencies. With the adoption of a new curriculum, there was the qualitative growth of the teachers and students' body, and an increase in the autonomy of the school to set out and update curricular components. PMID- 15782719 TI - [Hilda Anna Krisch: a nursing pioneer in Santa Catarina--education and contribution for the profession]. AB - This is a socio-historical research whose objective is to unveil the historicity of the first nurse in Santa Catarina who graduated at a Nursing School in the twentieth century, a character who had a prominent role in the national scene for her performance as a nurse. The data collection was carried out through the examination of many historical documents: interviews conducted with her and other people who had been part of the scene of the time, as well as other documentary sources, such as letters, school and professional reports, pictures, among others. To analyse the data, it was used an approach based on the social imagery. The data allowed us to identify her origins and path as a nurse, as well as the activities performed during her professional life. It can be concluded that Hilda Anna Krisch was a leader in the field not only in Santa Catarina but also in Brazil due to her role inside and outside her professional specialty. PMID- 15782721 TI - Doolin Lecture 2004. Alcohol policy serving our economy or society. PMID- 15782720 TI - [The education that we have and the one we want: a perspective on discourses]. AB - This research has the purpose of gaining more understanding about the actions and policies put in place in order to ensure the training of professionals that work in the services of the State Health Care System, considering the direct and intrinsic relation between the education and its final quality. Broken down into 3 perspectives, here labeled as voices, we have tried to identify, by means of the discourses on social control in the area of health, the system's administration, and education represented by the schools of nursing, what are the criticisms and propositions concerning the theme of professional training for the State Health Care System. The text deals with the terms used in the context of professional training, as well as a critical reflection about the new tendencies and paradigms that are on the agenda in current debates and discussions. PMID- 15782722 TI - Upgrading intensive care units--getting the design right prevents infection. PMID- 15782723 TI - Birth-weight trends in primiparous women in the twentieth century. AB - We investigated the trend in birth-weights in the Rotunda hospital in primiparous women over the 20th century. There was a significant increase in birth-weight particularly in the latter third of the century. In light of these findings we believe that new approaches to the management of labour in primiparous mothers be developed. PMID- 15782724 TI - Parents' knowledge of and attitude towards the primary childhood immunisations. AB - Parental knowledge of immunisation is essential to assist public health initiatives to increase the uptake rates. A cross-sectional survey design was employed to ascertain the knowledge and attitude of the primary childhood immunisations in randomly selected parents in a deprived inner-city area; 82% of 93 responded. Parents' reported uptake of immunisation was compared with general practitioner and official health board records. Respondents with a low standard of education (70% of respondents without a Leaving Certificate versus 44% with the Leaving Certificate p=0.06) and parenting alone (69% versus 49% of two-parent families p=0.07) were less aware of the severity of the diseases. Older parents (58%) questioned immunisation safety more than younger parents (28%), p=0.06. Full immunisation was reported by 91% of parents compared to 81% from the general practitioners records and 70% from health board figures. The poor correlation between parents' reported immunisation uptake and official immunisation sources could be improved through the use of personal identification numbers. PMID- 15782725 TI - Acute stroke care in an Irish general hospital. AB - A majority of in-patients with acute stroke in Ireland are cared for by general physicians. We studied the process of care and outcomes of stroke patients admitted to an acute general hospital in a rural setting. Ninety-five patients (55 males and 40 females) were admitted over 36 weeks. A majority of patients had a CT brain scan (97%) and were assessed by the multidisciplinary team (59-70%). The mean length of stay was 16.6 days. In patient mortality was 18.9%. The commonest discharge destination was to the Medical Rehabilitation Unit (44%). A high prevalence of cardiovascular risk factors was identified indicating significant potential for secondary prevention. PMID- 15782726 TI - Training and manpower issues for specialist registrars in paediatrics. How are we doing and where are we going? AB - All Irish paediatric higher specialist trainees' opinions about the paediatric higher specialist training (HST) scheme and related manpower issues were surveyed. Information was obtained on 1) trainees' level of satisfaction with HST, 2) their ultimate career ambitions including location of final posts, 3) attitudes to both flexible training and consultancies and 4) demographics to assess the significance of gender variations. Fifty-two eligible trainees were identified using the Royal College of Physicians of Ireland database. The survey was administered as an anonymous postal survey. The response rate was 88%. Results indicated a high level of satisfaction with HST (78%) overall although problems were noted with the half-day release programme as only 63% were facilitated. Only 30% wish to practice as subspecialists, 76% of trainees wish to work in an urban hospital and 43.5% desire a flexible consultancy suggesting an incompatibility of trainees' desires with current Irish medical manpower policy. To address these difficulties we suggest establishing more rigorous audit of training posts to ensure deficiencies are corrected and the establishment of flexible training to address gender imbalance and to promote the concept of consultant job sharing. PMID- 15782727 TI - Inadequacies in the first aid management of burns presenting to plastic surgery services. AB - Burn injury is a significant cause of mortality and morbidity in Ireland. Simple first aid measures such as immediate wound cooling and removal of the source of injury can significantly improve clinical outcome. We conducted a six-month study, investigating the first aid measures taken by both patients and their primary carers following burn injury. Of the 63 patients referred to the plastic surgery services, only 23.2% had employed the correct first aid principles. Only 20% of patients who sustained burns at work reported that first aid was available in the workplace. This study illustrates that knowledge regarding the initial management of burn injury is very poor. We suggest that a national public health education campaign could have a positive effect on the outcome of burn injury. PMID- 15782729 TI - Medical graduates of the National University of Ireland in 1978: who and where are they? AB - To inform debate on medical manpower planning and aspects of medical education, we gathered data on graduates of three Irish medical schools in 1978. Twenty six years later, four of the 236 graduates had died and seven were untraceable. All but one of the remainder were in clinical practice and in a wide range of disciplines. A third were overseas. The implications of these findings are briefly discussed. PMID- 15782728 TI - A retrospective review of visual outcome and complications in the treatment of retinoblastoma. AB - The aim of this study was to look at the visual outcome and treatment complications of children diagnosed with Retinoblastoma during the years 1985 2003 inclusive. A retrospective review of all patients records was performed. Patient characteristics, treatment methods and complications were recorded. Twenty eight children presented to Temple street Hospital between 1985-2003. Six of these infants had bilateral tumours. The mean age at presentation was 23.7 months. Sixty-nine percent presented with Leucocoria, of these 33% also had a squint. The mean duration of symptoms was only known in 58% and this figure was approximately 19.8 months. Enucleation was performed in 24 eyes of 24 patients. Three patients required adjuvant chemotherapy post enucleation. Two eyes was treated with external beam radiation and one eye with plaque radiotherapy. One eye (second eye) was treated with systemic chemotherapy and radiation. Five eyes of three patients were treated with systemic chemotherapy followed by adjuvant Argon laser, cryotherapy and diode laser to each eye.The complications of each treatment group was recorded. The visual outcome in the salvaged eyes was favourable. There were no deaths recorded. Though chemotherapy with adjuvant local treatments provide adequate treatment for early tumours, enucleation still plays a major role in the treatment of Retinoblastoma. The total eye salvage rate in this study was 29% with an enucleation rate of 90% in unilateral cases and 33% in bilateral cases. Sixty-six percent of bilateral eyes affected were salvaged. Seventy-one percent of tumours were diagnosed after a parent noticed a gross abnormality of the eye. This highlights the possible need for screening for retinoblastoma in the infant population. PMID- 15782730 TI - Hip protectors--a survey of practice in Ireland. AB - When used appropriately and worn by the user, it has been established that hip protectors can reduce the risk of femoral fractures in older people who fall. While hip protectors receive increasing interest in the scientific literature, it is not clear how widely and appropriately they are being used in Ireland. This national survey aimed to investigate this practice. The survey yielded 185 responses from a variety of settings, response rate of 53%. It noted that only 18% of respondents actually used hip protectors for their clients/patients, despite the fact that the many of the participants in this survey were from nursing homes, where the evidence supports their use. It clearly identifies that a lack of knowledge about hip protectors is one barrier to their use, with 54% of participants reporting they were not aware of hip protectors and their value; but other factors, such as the suitability and compliance of older clients/patients and financial issues also impact on their use. In excess of 70% of respondents reported that financial restrictions influenced their prescription practice. While the majority of the participants were nurses, it is of note that the professional group most frequently involved in prescribing the use of hip protectors was physiotherapists and not nurses or doctors. National guidelines which both educate and advise on the use of hip protectors would lead to optimal use in the context of both clinical and cost effectiveness. PMID- 15782731 TI - Are hospital based notifications enough? Achieving accurate case ascertainment- the Southern Ireland Cerebral Palsy Register (SICPR). AB - The Southern Ireland Cerebral Palsy Register (SICPR) attempts to identify all cases of cerebral palsy in the Southern Health Board region. Multiple sources of case ascertainment are necessary to achieve a complete register. The aims of the study were to ensure the accuracy of case ascertainment to this register. A postal survey to all general practitioners in the Southern Health Board region did not confirm any new cases of cerebral palsy to the register. We encourage all health professionals to notify our register of any child with suspected cerebral palsy. PMID- 15782732 TI - Introducing laparoscopic management of ectopic pregnancy to a busy gynaecology unit--is it worthwhile? AB - A review was carried out on 26 consecutive women undergoing surgery for ectopic pregnancy in the Limerick Regional Hospital, Ireland from April 2000 to April 2002. 13 were managed laparoscopically and 13 had laparotomy. There were no significant differences in age, parity or gestational age. 3 patients had previous ectopic pregnancy. 12/13 from the laparotomy group had a diagnostic laparoscopy prior to laparotomy. Anaesthetic time differed by 21.2 minutes with laparotomy being done faster than the laparoscopy group while operative time was 7.3 minutes longer in the laparoscopic group. The laparoscopic approach was associated with lower intraoperative blood loss (<50 ml vs 413.1 ml), less post operative analgesia requirement, shorter hospital stay (2.4 days vs 4.5 days), faster return to work (2 weeks vs 4 weeks) and less subsequent wound infection. Operative laparoscopy also has the advantage of being a diagnostic as well as a therapeutic tool in one procedure. PMID- 15782733 TI - Spontaneous rupture of the liver following a normal pregnancy and delivery. AB - Liver haematoma and rupture is a rare but potentially devastating complication of pregnancy. The majority of cases are associated with severe preeclampsia. We report a case of spontaneous hepatic rupture in a 39-year old woman following a normal pregnancy and delivery. Shoulder tip pain and serum haemoglobin of 6.8 prompted immediate imaging with computed tomography. A large subcapsular haematoma involving the right lobe of the liver was identified and managed conservatively. This diagnosis should be considered in the patient who presents with pain in the upper part of the abdomen and signs of haemorrhagic shock, even in the case of an uncomplicated pregnancy. PMID- 15782734 TI - Maturity Onset Diabetes in the Young (MODY) type 3. PMID- 15782735 TI - Extremely high PSA (250ng/ml) in a patient with localised prostatic carcinoma. PMID- 15782736 TI - Tackling abuse requires awareness, accountability, action. PMID- 15782737 TI - Secret filming of poor care leads to NMC investigation. PMID- 15782739 TI - Challenging restraint. PMID- 15782738 TI - A word from Wales. PMID- 15782740 TI - New solutions for ageing problems. PMID- 15782741 TI - Teaching people with Parkinson's disease about their medication. AB - PD is a progressive neurodegenerative disorder affecting an estimated 78,000 Australians. Predominantly it affects older people, although or younger. Medications to treat PD are aimed at controlling symptoms as there is no known cure. A regime of PD medications may involve taking doses at frequent intervals and adverse reactions are common. Education of the person with PD, their close carers and family that covers medications, observational techniques and dealing with side effects, along with unpredictable worsening of PD symptoms, may result in improvement in quality of life. When the person with PD has realistic expectations about PD drug therapy then the ability to cope with the physical disability caused by symptoms may be improved. It is important to educate about any possible interaction with commonly taken drugs including dietary supplements and over-the-counter medication. Before a medication education session, nurse activities should include assessment of the person with PD and their support network, development of an appropriate education plan and subsequent implementation, followed by evaluation. Ongoing nursing support at each clinic visit can provide opportunity for evaluation. An individualised session guided by the principles of adult learning theory may provide a successful tool for use in the education of people with PD as well as healthcare professionals. A large amount of available information and untested educational material does not address the educational needs of people with PD with regard to their medications. There is a need for further research in this area. The effectiveness of a one-on one educational session about PD medication supported by printed, individualised PD medication information may provide more appropriate education and lead to improved quality of life. It is the focus of a planned research study. PMID- 15782742 TI - Implementing a best practice statement in nutrition for frail older people: Part 2. PMID- 15782743 TI - Dementia and challenging behaviour: the needs of family caregivers. PMID- 15782744 TI - Older people with learning disabilities. Part 2: Accessing care and the implications for nursing practice. AB - In the second of two articles, Robert Jenkins concludes his discussion of nursing care for older people with learning disabilities by considering some barriers to care and the implications for nursing practice. Part 1 discussed individuals, ageing and health. PMID- 15782745 TI - Promoting healthy sight and eye care. PMID- 15782746 TI - The common touch. PMID- 15782747 TI - From rough sleepers to royalty. PMID- 15782748 TI - No bars to better care. PMID- 15782749 TI - Will high-quality care be the norm by the time I need it? PMID- 15782750 TI - Clinical relevance of automated drug alerts from the perspective of medical providers. AB - The authors used a real-time survey instrument and subsequent focus group among primary care clinicians at a large healthcare system to assess usefulness of automated drug alerts. Of 108 alerts encountered, 0.9% (n = 1) represented critical alerts, and 16% (n = 17) were significant drug interaction alerts. Sixty one percent (n = 66) involved duplication of a medication or medication class. The rest (n = 24) involved topical medications, inhalers, or vaccines. Of the 84 potentially relevant alerts, providers classified 11% (9/84), or about 1 in 9, as useful. Drug interaction alerts were more often deemed useful than drug duplication alerts (44.4% versus 1.5%, P < .001). Focus group participants generally echoed these results when ranking the relevance of 15 selected alerts, although there was wide variance in ratings for individual alerts. Hence, a "smarter" system that utilizes a set of mandatory alerts while allowing providers to tailor use of other automated warnings may improve clinical relevance of drug alert systems. PMID- 15782751 TI - Getting physicians to make "the switch": the role of clinical guidelines in the management of community-acquired pneumonia. AB - The authors sought to assess physician awareness and usage of American Thoracic Society guidelines for early conversion from intravenous to oral antibiotics ("switch therapy") in those with community-acquired pneumonia (CAP). We then determined if adoption of a CAP guideline would improve either. Patients (N = 510) hospitalized with CAP from June 2002 to May 2003 were identified retrospectively, and chart reviews were done on a random sample (130 [25%]) of these. Physicians were surveyed before and after guideline adoption. Community acquired pneumonia guideline implementation increased physician awareness of American Thoracic Society recommendations (5% to 40%) and use of switch therapy (60% to 86%). Such use resulted in decreased overall length of stay from 3.6 to 2.4 days (P < .05) and from 2.91 to 2.41 days (P < .05) among early-switch candidates. Early-switch therapy was not optimally used prior to implementation of this CAP guideline. Adoption of the guideline increased awareness and reduced length of stay among inpatients with CAP. PMID- 15782752 TI - Factors related to physicians' adoption of electronic prescribing: results from a national survey. AB - Electronic prescribing (E-RX) is a component of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA). The objective of this study was to identify factors related to physicians' adoption of E-RX for outpatients. This study employed an electronic survey of US physicians who subscribe to the Physicians Online Internet service. Electronic prescribers were compared to traditional prescribers in terms of demographics, practice type and location, technology use, and beliefs about E-RX. A total of 1104 physicians responded, 19% of whom prescribed electronically. Electronic prescribers were more likely to be generalists practicing in academic or publicly funded centers, have fewer years in practice, and work in technology-equipped offices. They also held different beliefs versus traditional prescribers in terms of E-RX limitations and its potential to improve medication safety and prescribing efficiency. In addition to financial incentives established by MMA, adoption can be stimulated by improvements in the technology and on organizational commitment. PMID- 15782753 TI - Smokers' interest in quitting and services received: using practice information to plan quality improvement and policy for smoking cessation. AB - Given the prevalence of smoking, its impact, and the benefits of cessation, helping smokers quit should be a top priority for health care organizations. To restructure health care delivery and guide future policy, the authors used baseline survey data from an 18-site Veterans Health Administration group randomized trial to assess the level of interest in quitting smoking for a practice population and determine what smoking cessation services they reported receiving. Among 1941 current smokers, 55% did not intend to quit in the next 6 months, and the remainder intended to quit in the next month (13%) to 6 months (32%). Forty-five percent reported a quit attempt in the prior year. While nearly two thirds of smokers reported being counseled about cessation within the past year, only 29% were referred to a cessation program, and 25% received a prescription for nicotine patches. Tobacco control efforts within this population should focus on increasing the rate of assisting patients with quitting. PMID- 15782755 TI - [The place of surgicenter nurse in the Third Millennium Ophthalmology]. PMID- 15782754 TI - Case study: development of and stakeholder responses to a nursing home consumer information system. AB - California Nursing Home Search (www.calnhs.org), launched in October 2002, provides information about nursing home quality to a broad range of stakeholders. This case study discusses the process of developing a consumer-oriented nursing home Web site and presents an analysis of postlaunch responses from a number of sources (i.e., media, outreach, Web site use, correspondence, meetings, interviews) to determine the impact of the site and how it can be improved and used as an example. Consumers found the Web site valuable, but some needed clarification on navigation. Providers had complaints about the use of quality ratings and concerns about public availability of the data. Most discharge planners and care managers do not use Internet resources to find facilities. Feedback, modifications, updates, and outreach are needed on a continuous basis to ensure the site is a helpful tool for all stakeholders. PMID- 15782756 TI - [The place of photodynamic therapy in the treatment of age-related macular degeneration]. AB - Age-related macular degeneration (ARMD) is a multifactorial disease, with unknown etiology, which represents one of the main causes of legal blindness. At this moment, the only treatment for ARMD is symptomatic and concerns the distinction of the choroidal neovascularisation (CNV). There are many therapeutical procedures, and their classification is difficult. The dynamic phototherapy (PDT) with intravenous injection of verteporfirin (Visudyne), a recent therapeutic concept based on a photochemical reaction, represents the only standard procedure for destroying the macular CNVs. The paper presents new therapeutical procedures, discussing them in relation with PDT. Beside the PDT and its supporting methods, other competing techniques (transpupillary thermotherapy, surgical removal, anti angiogenesis and antioxidants) are presented. PMID- 15782757 TI - [Ocular changes in diabetes mellitus (without diabetic retinopathy)]. AB - The paper propose to point out the modifications in the conjunctiva, cornea, iris, ciliary body, lens, vitreous body and choroid due to the diabetic disease. The alterations were mainly demonstrated by optic and electronic microscopy. It has been also studied the relationship between the type and the severity of this lesions, the stages of the diabetic retinopathy and the glycemic level. In conclusion, the complete ophthalmologic examination of diabetic patients can provide extremely useful information for the therapeutical management and visual prognosis. PMID- 15782758 TI - [Latanoprost topical ophthalmic combinations]. AB - Latanoprost became the first line therapeutic agent in glaucoma treatment being the best sold worldwide anti-glaucoma medication. When adequate intraocular pressure decrease can not be achieved with latanoprost monotherapy, then its combinations are to be used. Latanoprost combinations are grouped in to non-fixed and fixed variants. Non-fixed combinations mean concomitant therapy,that is giving the two or more medications using different bottles. The medications used for latanoprost non-fixed combinations are represented by timolol 0.5%, pilocarpine 2%, acetazolamide dispensed systemically and locally, dipivefrin 0.1%, unoprostone 0.12% and brimonidine 0.2%. Fixed combinations mean administering the two mixed medications using one single bottle. At the present time there is only one fixed combination of latanoprost i.e. its combination with timolol 0.5% whose trading name is Xalcom. PMID- 15782759 TI - [Primary open-angle glaucoma associated with pigmentary epithelium tumor of the iris]. AB - The authors present a case of a 63 years old patient (the case was published in Rev. Oft. No.2/2001, vol. XLXII) which is under a 9 years treatment already for open angle glaucoma. In addition the patient presents pigmentary epithelial iris tumor. This case was studied for 3 years. PMID- 15782760 TI - [Vascular changes in optic disk drusen]. PMID- 15782761 TI - [Phacoemulsification in posterior polar cataract]. AB - PURPOSE: To evaluate the technical particularities of phacoemulsification and the intraoperative complications in posterior polar cataract. MATERIAL AND METHODS: Prospective study which comprises 10 eyes with posterior polar cataract (8 patients), operated on between May 2001 and April 2004 in the ophthalmology department of the Military Hospital Cluj-Napoca, by the same surgeon. Phacoemulsification with low parameters was performed in all cases, and acrylic and PMMA intraocular lenses were implanted in 9 eyes. RESULTS: The patients' age was between 24 and 80 years, with a mean age of 55.12 years. The majority of the patients presented bilateral posterior polar cataract (70%), but only 2 patients were operated on the both eyes in the analyzed period. The posterior capsule rupture occurred in 4 eyes (40%). A posterior residual plaque was attended in one case, but did not influence the short term visual acuity. The visual acuity of the 3 cases which were affected unilaterally was not influenced by the surgery, which was without complications in 2 cases. The causes were preexisting amblyopia. CONCLUSIONS: The study confirms the high risk to posterior capsule rupture during posterior cataract surgery and the high incidence of the bilateral form (70%). PMID- 15782762 TI - [Keratitis as an emergency in the public service at the Hotel-Dieu Hospital in Paris]. PMID- 15782763 TI - [Phacoemulsification and pseudoexfoliative syndrome]. AB - PURPOSE: To compare the results and the complications of cataract surgery by phacoemulsification in eyes with pseudoexfoliation syndrome (PEX) and without pseudoexfoliation syndrome. METHODS: A retrospective clinical study (January 2001 - January 2004; mean follow-up 21.5 months) of 116 cases divided in two groups: group A (55 eyes) with senile cataract and pseudoexfoliation syndrome and group B (61 eyes) with senile cataract but without pseudoexfoliation syndrome. The eyes were without another associated ophthalmological diseases and the phacoemulsifications were performed by the same surgeon. Intraoperative observations included anterior chamber depth, pupil size after maximal mydriasis, zonular dehiscence, capsule tear, vitreous loss and postoperatively: best corrected visual acuity, intraocular pressure (IOP), aqueous flare response and IOL stability were evaluated at 1 day, 1 week, 1 month and until 3 years. The Student-t test was used for statistical analyses. RESULTS: There were determined significant correlations between pupil size, anterior chamber depth and the rate of complications in the group A. The visual acuities were similar in both groups. IOP reduction was superior in the eyes with PEX as compared with control group, but without statistically significantly difference. In group A, there were a non statistically significantly percentage of IOL dislocation as compared with group B. In the eyes with PEX, it is necessary to utilize IOLs with bigger optic size. CONCLUSIONS: Phacoemulsification represents a safe method in cataract surgery in eyes with pseudoexfoliation syndrome in the presence of careful surgical technique: maximal mydriasis, circular continuous curvilinear capsulorhexis, adequate hydrodissection. On the long run, the postoperative results were comparable between eyes with PEX and the control group (without PEX). PMID- 15782764 TI - [Clinical and experimental study of the effect of blue dye on corneal endothelium]. PMID- 15782765 TI - [Clinical and serologic research in toxoplasmic uveitis in children]. AB - PURPOSE: We have done a retrospective study of juvenile toxoplasmic uveitis cases hospitalized between 1993-2002. MATERIAL AND METHOD: 96 juvenile uveitis were available and 21 cases were diagnosed with juvenile toxoplasmic uveitis. RESULTS: In this study we have analyzed the distribution of cases per year, each of them was investigated by functional and objective ocular examinations, laboratory tests and treatment. PMID- 15782766 TI - [Cataract surgery in Romania--2003]. AB - AIM: to evaluate cataract surgery performed in Romania analyzing: --the number of cataract surgery performed in 2003; --the surgical technique used: intracapsular extraction, extracapsular extraction, phacoemulsification; --the number and type of intraocular lenses used; --the number of ophthalmologists that performed cataract surgery; MATERIAL AND METHOD: --a questionnaire was sent to all ophthalmologists which do cataract surgery. RESULTS: --38,000 surgeries for cataract were performed in 2003; --the surgical techniques used were : Phacoemulsification (18,000 cases), classic extracapsular extraction ( 19,000 cases) and intracapsular extraction (1000 cases); --28,500 rigid and 8,000 foldable intraocular lenses were implanted; - 223 ophthalmologists perform cataract surgery in Romania. PMID- 15782767 TI - [The Bioptron light therapy]. AB - The Bioptron light therapy system acts naturally, upholding the capacity of regeneration of the body. Since the discovery of the therapeutical effects of the Bioptron light, over 20 years ago, its use as treatment has been developed for a large variety of diseases, among which also the eye-diseases (simplex and zoster herpes, conjunctivitis). PMID- 15782768 TI - [Correction of refractive errors in children with contact lenses]. AB - PURPOSE: To appreciate the indications, complications and visual outcomes in the correction of refractive errors with contact lenses at children. MATERIAL AND METHOD: We studied 15 eyes from nine kids. The times prescription included: history (motivation of wearing contact lenses), ocular refraction (with cycloplegia), best corrected visual acuity, keratometry, the corneal diameter, slit lamp examination, the selection of contact lens and follow up. RESULTS: The age of patients varied between several months to twelve years. The refractive errors in which we used contact lenses were: aphakia--6 eyes, myopia--6 eyes, hyperopia--3 eyes. The types of contact lenses were for permanent use--8 eyes and discontinuity use--7 eyes. We didn't have any major complication. The best corrected visual acuity with contact lenses was: 0.1-0.3 at 4 eyes, 0.4-0.6 at 2 eyes and one at 7 eyes. CONCLUSIONS: (1) Contact lenses represent the optimal correction of anisometropia after congenital or traumatic cataract operation in children. (2) The quality of vision obtained with contact lenses represents an important factor in the prevention and treatment of amblyopia. PMID- 15782769 TI - [Explantation and replacement of intraocular lenses]. AB - PURPOSE: to evaluate the reasons for explantation of different types of intraocular lenses. MATERIAL AND METHOD: We studied 27 eyes operated previously for cataract and which developed in postoperative period intraocular induced complication, demanding explantation. RESULTS: In 51.85% cases we explanted anterior chamber IOL, in 40.74% eyes posterior chamber IOL and in the rest pupillary IOL. Reasons of explantation were according to the artificial lens: in 4 cases for edematous keratopathy, chronic uveitis in 2 eyes and descentration in 8 eyes. Reimplantation was performed at 19 eyes, in 10 cases with standard posterior chamber IOL and in 9 cases with scleral sutured posterior chamber IOL. CONCLUSIONS: The main reason for explantation in anterior chamber IOL is the edematous keratopathy and malpositions for posterior chamber IOL. PMID- 15782770 TI - [Indications of enucleation]. PMID- 15782771 TI - [Biomechanical stress in glaucoma--cause and effect]. AB - Glaucoma is a neurodegenerative disease with multifactor causes which is characterized by acute loss of ganglion cells. Many mechanisms are involved in this disease but they are incomplete elucidated. Biomechanical stress generate changes to lamina cribrosa which determine ganglion cells death. STUDY OBJECTIVES: The quantification of changes of LC and peripapillary sclera in experimental models with glaucoma. METHODS AND MATERIAL: A prospective, experimental study (10 weeks) using 8 adult guinea pigs in which glaucoma was induced by equatorial burns (diathermia) at two episcleral veins--right eye, left eye being considered witness. The monitored parameters are: IOP, retro-ocular blood flow, histopathologic examination. RESULTS AND CONCLUSIONS: Constant exposure to biomechanical stress (increase to IOP) involve to local level cellular and molecular mechanisms which made changes in the extracellular matrix. The changes of LC are a mechanism that determine glaucomatous optic nerve disease and became a new option to diagnose and follow in glaucoma. PMID- 15782772 TI - Endophytes--the chemical synthesizers inside plants. AB - Endophytes are microbial entities that live within living tissues of plants. In most cases their relationship with the host plant is symbiotic and probably mutualistic. Many are capable of synthesizing bio-active compounds that can be used by the plant for defense against pathogenic fungi and bacteria. Some of these compounds have proven usefulfor novel drug discovery. By encouraging the endophytes to grow outside the plant in nutrient rich media, it is possible to harvest the bio-active compounds that they produce. However, much needs to be discovered and understood about the host/endophyte relationship before we can fully utilize endophytes in the discovery of medicinally important compounds. PMID- 15782773 TI - Solar transient events and their importance for coronal heating. AB - Over the last decade, the uninterrupted high resolution coverage of the Sun both from the excellent range of telescopes aboard many spacecrafts and from ground based instruments has led to a wealth of observations of small-scale dynamic events observed from the chromosphere to the transition region and corona. Since many of these events were observed with different instruments they show different properties from one another. It is suggested that many of these phenomena result from small-scale reconnection events due to the continually evolving magnetic fields as seen at the photospheric level, although waves are thought to play an important heating role as well. Nowadays, there is a general consensus that the key to understanding how the solar plasma is accelerated and heated may well be found in the study of these small-scale dynamic events. Here we give a brief review of the range of observed transient features and suggest that these small scale events may well have broad implications for the mass and energy balance of the whole upper atmosphere of the Sun and the solar wind. PMID- 15782774 TI - Hematological and iron-related analytes--reference data for persons aged 1 year and over: United States, 1988-94. AB - OBJECTIVES: This report presents national estimates of hematologic and iron related analytes for persons 1 year of age and over, by age, sex, and race/ethnicity. METHODS: The analysis is based on data from the third National Health and Nutrition Examination Survey (NHANES III) (1988-94), which was designed to provide information on the health and nutritional status of the civilian noninstitutionalized U.S. population. The sample used for these analyses included the 26,372 participants who had laboratory tests. RESULTS: This report provides mean, standard error of the mean, median, and percentile laboratory values for the U.S. population, 1988-94, for hematological and iron-related analytes. In addition, percentage distributions are provided. CONCLUSIONS: Data on hematological and iron analytes provide reference values for clinical and longitudinal comparisons. PMID- 15782775 TI - [Historical outline of forensic medicine in Poland and its connections with forensic medicine in German-speaking countries]. AB - The author briefly presented development of forensic medicine as a separate branch of medical sciences in Poland from the end of the Middle Ages to date. He comments on conditions in which forensic medicine developed after partitions of Poland and its restoration of independence. He emphasized the merits of Prof. Wachholz and his pupils, as well as the great publishing effort of forensic scientists during the interwar period and after World War II. He did not fail to highlight links between Polish and German forensic medicine and quotes numerous examples of very close contacts between both communities in this field. PMID- 15782776 TI - [50th anniversary of the Department of Forensic Medicine, Medical University of Bialystok (1954-2004)]. AB - The paper illustrates the history, scientific and teaching achievements of the Department of Forensic Medicine, Medical University of Bialystok in commemoration of its 50th Anniversary. PMID- 15782777 TI - [Prof. Maria Byrdy--Doctor Honoris Causa AMB 1990, the founder of the Chair of Forensic Medicine, the first head of the Department of Forensic Medicine, Medical University of Bialystok in 1954-1984]. AB - The paper portrays the scientific, publicist and teaching activities of Prof. M. Byrdy, with special regard to her residence at the Departments of Forensic Medicine in Cracow (1939-1953) and Bialystok (1954-1984). PMID- 15782778 TI - [Death in a duel: historical penal and medicolegal aspects of a case from 1903 as a background]. PMID- 15782779 TI - Population studies for STR loci (D3S1358, D5S818, D7S820, D18S51 and FGA) in NWFP and Sindhi populations of Pakistan for forensic use. AB - CEMB's Forensic DNA typing project is directed towards the introduction of DNA typing technique in Pakistan's criminal justice system so as to exonerate an innocent and wrongly accused person and incriminates the culprit. The present study is a part of the project of CEMB to analyze Sindhi and NWFP (North West Frontier Province) populations for five STR (Short Tandem Repeat) loci out of 13 CODIS (Combined DNA Index System) loci. Allelic frequencies and heterozygosity for STR markers D3S1358, D5S818, D7S820, D18S51 and FGA (FIBRA) were determined. Samples from unrelated individuals were amplified by multiplex PCR using the unlabelled primers for these markers followed by denaturing Polyacrylamide Gel Electrophoresis (PAGE). Statistical analysis was performed to determine the allelic frequencies and was evaluated using the Chi Square Test. PMID- 15782780 TI - [Chemical diagnosis of carbamazepine intoxication and its usefulness in toxicological jurisprudence]. AB - Intoxication with carbamazepine (Amizepine, Tegretol) represents a significant problem of clinical and forensic toxicology, which frequently develops independently of the commonly used monitored therapy. In the study a technique of qualitative and quantitative analysis applied to carbamazepine (CBZ) as well as its metabolites carbamazepine 10,11-epoxide (CBZ-E) and 10,11 dihydroxycarbamazepine (CBZ-DH) in biological material using high-performance liquid chromatography (HPLC) was presented. The studies also involve isolation of studied compounds using solid phase extraction, SPE-Abselut NEXUS (Varian), establishment of calibration range and validation of analytical parameters. This was followed by a discussion of results obtained in routine diagnostic investigations in cases of intoxications, corroborating practical suitability of the designed technique. PMID- 15782781 TI - [Accidents in the workplace caused by alcohol intoxication]. AB - Fatal accidents in the workplace can be caused by work conditions, aggravation of a chronic disease or alcohol intoxication. The purpose of this paper was to show the influence of ethyl alcohol on accidents in the workplace with regard to the occupation and age of the examined individuals. A group of victims whose deaths resulted from other external factors (suicide, poisoning by non alcoholic agents, etc.) was separated and not included. Statistical analysis of the autopsies carried out in the Forensic Medicine Department, Silesian University of Medicine, Katowice in the years 1992-2001 showed that accidents in the workplace amounted to 4-6% of the total number of deaths in the space of a year with alcohol (ethanol) being the causative factor in 3-15%. PMID- 15782782 TI - [Long-term realistically life-threatening disease]. AB - In this paper the authors discuss a legal code description of a wrongly included (as a result of a legislative error) "long-term really life-threatening disease" (dlugotrwala choroba realnie zagrazajaca zyciu) in the Criminal Code. This category of disease impossible to apply in practice since its terms "long-term" and "realistically life-threatening" are mutually exclusive--is nonetheless applicable to crimes committed from Sept. 1, 1998 to Dec. 8, 2003. In effect this causes a change in the qualification of certain acts in Art. 156 of the Criminal Code, to include those in Art. 157, Paragraph 1, and in some cases, even extending to acts named in Art. 157, Paragraph 2 of the Criminal Code. PMID- 15782783 TI - [Malpractice in internal medicine in the case material of the Forensic Medicine Department, Medical University of Lodz, Poland]. AB - Legal opinions in the area of internal medicine, opinionated by the Forensic Medicine Department, Medical University of Lodz, Poland in 2002, were selected for further analysis and assessment in the course of medical treatment. During this period opinions were given in 20 cases, most of which concerned cardiovascular, secondly respiratory, followed by, endocrine diseases. In half of the cases analyzed it was determined that medical treatment had been incorrect. All of the objectionable diagnostic and therapeutic procedures evidenced some form of decisional error. In most cases errors were made in the diagnosis, as a result of neglect in performing necessary examinations, or of erroneous interpretation of results obtained. PMID- 15782784 TI - [Biostatistical interpretation of DNA paternity testing in cases of closely related alleged and biological fathers]. PMID- 15782785 TI - [The problems concerning estimating age at death based on morphological features of adult skeletons]. AB - The paper contains a review of biological definitions of age and their application in forensic anthropology as a theoretical basis for age at death identification. The authors discuss the reasons and importance of discrepancies between chronological and developmental age. Also emphasized was the significance of "age at death" expression and paying attention to the limitations of age at death estimation. PMID- 15782786 TI - [Bibliography of Polish papers on forensic medicine, criminology and related fields published in 2003]. PMID- 15782787 TI - 'Burns' from anesthesia IV result in 500,000 dollar verdict. Owens v. Perez, 2005 WL 247537 S.W.3d-TX. PMID- 15782788 TI - Nurses refuse to follow orders: catastrophic results. Case on point. Szczuvelek v. Harborside Healthcare Woods Edge, 2005 WL 123829 A.2d-NJ. PMID- 15782789 TI - PA: Kicking motions made toward blind patient: firing for 'wilful misconduct'--UI benefits denied. Miller v. Unemployment Compensation Board of Review, No.2039 C.D. 2004 (Pa. Commw. 02/02/2005). PMID- 15782790 TI - Case on point: Sweetwater Hospital Association v. Carpenter, 2005 WL 240605 S.W.3d-TN. PMID- 15782791 TI - [Differential diagnosis of anemias]. AB - The authors describe the factors influencing of normal hemoglobin level, pathogenetic and morphological classification of anemias and the possibilities to distinguish their different types. They highlight features of iron deficiency anemia and anemia of chronic diseases. They summarise in tables the basis of laboratory diagnosis and possibility of identification of different types of anemias. PMID- 15782792 TI - [Right subaxillary and posterolateral thoracotomy for open repair of congenital heart defects]. AB - OBJECTIVE: Low-risk cardiac surgery approaches zero morbidity/mortality, therefore, cosmetic issues append. Right thoracotomy has been advocated as cosmetically attractive alternative to median sternotomy. Posterolateral thoracotomy involves chest wall musculature division that contributes to postoperative pain/morbidity; access to the heart may be difficult. Longitudinal subaxillary incision and muscle-sparing thoracotomy can overcome these disadvantages and provide better visibility. PATIENTS AND METHODS: Between April 2000 and April 2004 the authors performed open-heart repair from right thoracotomy in 161 patients. Diagnoses comprised perioval and sinus venosus ASDs (137), VSD (11), incomplete/intermediate AVD (8), miscellaneae (5). According to both patients' and surgeon's preference a longitudinal midaxillary incision and muscle-sparing lateral thoracotomy was utilised in 98 cases. Intracardiac repair was performed by extracorporal circulation, usual cannnulation, cardioplegia. Data of 79 ASD closures via midline sternotomy were used as controls. RESULTS: Sternotomy and thoracotomy groups did not differ in demographic characteristics albeit thoracotomies exhibited wider age and weight range. The skin incision could be easily stretched up by 60% providing excellent visibility/access. No mortality occurred. Phrenic nerve palsies (2), transient tetraparesis (1) fully recovered. A mitral valve replacement was necessary for valve pathology unrelated to access. No breast and/or upper limb vascular/neurologic problems were encountered. CONCLUSIONS: Right subaxillary muscle-sparing thoracotomy offers safe option for open-heart repair of selected anomalies. Despite the shorter skin incision subaxillary approach provides better visibility than conventional thoracotomy as mediastinal organs are closer to the operator. Intermediate cosmetic result are appealing. With no muscles divided less postoperative pain is anticipated. The authors recommend to expand the spectrum of anomalies repaired via this approach. PMID- 15782793 TI - [Modern diagnosis and treatment of pancreatic cancer--from the viewpoint of the internist]. AB - INTRODUCTION: Despite considerable progress in the areas of epidemiology and molecular genetics, pancreatic cancer is still characterized by a dismal prognosis. Pancreatic cancer remains a major unsolved health problem, with conventional cancer treatments having little impact on disease course. AIM: The authors purpose was to offer an overview of the modern diagnosis and treatment of pancreatic cancer for internists and every doctors who are interested in this subject. METHOD: The current literature has been reviewed and summarized. RESULTS AND CONCLUSIONS: Resectability rates and reduction in postoperative mortality are achieved by centralisation of treatment in high-volume speciality centres. For locally advanced, unresectable, and metastatic disease treatment is palliative, however, new methods are being investigated. The current overview provide recommendations for diagnostic and therapeutic algorithms. PMID- 15782794 TI - [Notch signaling in the regulation of hematopoiesis]. AB - Notch signaling defines an evolutionarily ancient cell interaction mechanism. The signals transmitted through the Notch receptor, in combination with other cellular factors influence differentiation, proliferation and apoptotic events at all stages of development. Recent advances have elucidated both the biochemical mechanism regulating receptor activation and the molecular participants forming the intracellular signaling cascade. Authors present description of the main signaling components involved in the Notch pathway and how it can affect the growth and function of lymphocytes. Notch signaling is critical during lymphocyte development, and dysregulation of the pathway can give rise to leukemia. It is conceivable that appropriate manipulation of Notch signaling may become a useful tool in addressing a variety of human dysplastic condition and tissue regeneration. PMID- 15782795 TI - [A case of absent right and persistent left superior vena cava: echocardiographic diagnosis]. AB - A 49-year-old man underwent transthoracic echocardiography for atypical chest pain. A dilated coronary sinus was found and venous contrast echocardiography raised the suspicion of absent right and persistent left superior vena cava. Transesophageal echocardiography showed no presence of right superior vena cava. The echocardiographic findings were confirmed by upper venous digital subtraction cavography. PMID- 15782796 TI - [Treatment of hyperthyreosis]. PMID- 15782797 TI - Perception is everything. An ACPE survey reveals the details of physicians' public fall from grace. Can they regain the credibility and trust they once enjoyed? AB - Once among the nation's most-respected professionals, doctors have fallen to No. 5--a step below grade-school teachers and tying with druggists. A survey by the American College of Physician Executives reveals that some docs are critically examining their own medical values and moral principles. Kenneth Goodman, left, director of the University of Miami's bioethics program, says the survey will prove that business as usual is not necessarily good business. PMID- 15782798 TI - Looking for transparency. Charities may face more stringent IRS requirements. PMID- 15782799 TI - No concessions. Medicaid cuts are likely but severity is unknown. PMID- 15782800 TI - Tenet's new tangle. Other states could follow Fla. with racketeering charges. PMID- 15782801 TI - Legislative long shot. Another universal coverage bill introduced in Calif. PMID- 15782802 TI - UHS eyes undervalued assets. PMID- 15782803 TI - Ante up. Louisiana's rural hospitals upset over unique fees. PMID- 15782804 TI - Critical time at rural hospitals. PMID- 15782805 TI - Medicare modernization at a cost. Still more billions put health plans on the hook to prove privatization works. PMID- 15782806 TI - A tough job, but I love it. Clear goals and a new culture have HealthSouth positioned for the future. PMID- 15782808 TI - Citizen reformers. Panel to address health system change. PMID- 15782809 TI - By the numbers. Largest healthcare mergers announced in 2004 ranked by deal size. PMID- 15782807 TI - Construction--and costs--going up. Even as expenses mount, pressured by rising commodity prices, healthcare building continues to boom, annual survey shows. PMID- 15782810 TI - On the front lines. PMID- 15782811 TI - Meet you at the joint. PMID- 15782812 TI - Healthcare. Let the discussion begin. PMID- 15782813 TI - Bioterrorism. It was unbelievable. PMID- 15782814 TI - Molecular components and mechanism of adrenergic signal transduction in mammalian pineal gland: regulation of melatonin synthesis. AB - Rhythmic neural outputs from the hypothalamic suprachiasmatic nucleus (SCN), which programme the rhythmic release of norepinephrine (NE) from intrapineal nerve fibers, regulate circadian rhythm of melatonin synthesis. Increased secretion of NE with the onset of darkness during the first half of night stimulates melatonin synthesis by several folds. NE binds to both alpha1- and beta-adrenergic receptors present on the pinealocyte membrane and initiates adrenergic signal transduction via cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) generating pathways. The NE-induced adrenergic signal transduction switches 'on' melatonin synthesis during the early hours of night by stimulating expression of the rate-limiting enzyme of melatonin synthesis, N-acetyltransferase (AA-NAT) via cAMP-protein kinase A (PKA)-cAMP response element binding protein (CREB)-cAMP response element (CRE) pathway as well as by increasing AA-NAT activity via cAMP-PKA-14-3-3 protein pathway. Simultaneously, adrenergically-induced expression of inducible cAMP early repressor (ICER) negatively regulates aa-nat gene expression and controls the amplitude of melatonin rhythm. In the second half of night, increased release of acetylcholine from central pinealopetal projections, inhibition of NE secretion by SCN, withdrawal of adrenergic inputs and reversal of events that took place in the first half lead to switching 'off' of melatonin synthesis. Adrenergic signal transduction via cGMP-protein kinase G (PKG)-mitogen activated protein kinase (MAPK)-ribosomal S6 kinase (RSK) pathway also seems to be fully functional, but its role in modulation of melatonin synthesis remains unexplored. This article gives a critical review of information available on various components of the adrenergic signal transduction cascades involved in the regulation of melatonin synthesis. PMID- 15782815 TI - Preparation and in vitro evaluation of liposomal/niosomal delivery systems for antifungal drug clotrimazole. AB - Clotrimazole, an imidazole derivative antifungal agent is widely used for the treatment of mycotic infections of the genitourinary tract. In order to develop alternative formulation for the vaginal administration of clotrimazole to provide sustained and controlled release of appropriate drug for local vaginal therapy, liposomes/niosomes were evaluated as delivery vehicles. To optimize the preparation of liposomes/niosomes with regards to size and entrapment efficiency, multilamellar liposomes/niosomes containing drug were prepared by lipid hydration method. The ability of the systems to deliver clotrimazole into and through the mucosa was evaluated in vitro using rabbit vaginal mucosa with vertical Franz diffusion cells. The in vitro permeation data showed that the liposomes/niosomes system increased the clotrimazole total penetration through the vaginal mucosa by 1.6, 1.5-fold, the accumulation of clotrimazole into the mucosa was increased by 3.1, 2.3-fold, respectively, as compared with control during 24 hr. These results suggest that the studied liposomes/niosomes systems may be appropriate vesicles for the vaginal mucosa delivery of clotrimazole for local vaginal therapy. PMID- 15782816 TI - Effects of deltamethrin on granule cell migration during postnatal development of rat cerebellum. AB - Deltamethrin (DLT; 0.7mg/kg/body wt/day, i.p., dissolved in propylene glycol) administration during postnatal days 913 in Albino rat pups, resulted in a delayed appearance of radial glial fibers, that guide the migration of granule cells. Moreover, the radial glial fibers in the DLT-treated pups were disorganized, hypertrophied and heavily stained. Thus, it is being proposed that although after exposure to DLT the neuronal proliferation occurs at normal rate, the neuronal migration along the stumpy and crumpled radial fibers hamper the journey of the healthy neurons to their proper destination. PMID- 15782817 TI - Protective efficacy of maternal antibodies induced by Salmonella toxoid (vaccine). AB - An attempt was made to evaluate the protective efficacy of maternal antibodies in chicks against salmonellosis. Layer chicks ageing 21 days were individually vaccinated with 100 microg of Salmonella enterica subspecies enterica serovar Weltevreden (BM 1643) toxoid adjuvanted with vitamin E subcutaneously. After 90 days of the primary vaccination the birds were given booster dose of the vaccine. The saline extract of the yolk of eggs laid by the vaccinated birds yielded agglutination and ELISA titres ranging from 43.2 +/- 5.33 to 75.2 +/- 6.26 and 4.987 x 10(3) +/- 0.54 to 5.89x103 +/- 0.56, respectively. Sera of chicks hatched from eggs laid by the vaccinated layers were also subjected to agglutination and ELISA. Agglutination and ELISA titres on the 5th day--post hatching (dph) were 21.6 +/- 1.75 and 4.025 x 10(2) +/- 0.59, while on the 10th dph titers were 13.6 +/- 1.65 and 1.21 x 10(2) +/- 0.60, respectively. It was also observed that only one out of 6 chicks died when challenged with 2 x 10(9) CFU of S. serovar Gallirarum at the age of 7 days showing 83.33% protection. Thus it can be concluded that passive immunity confided by Salmonella enterica subspecies enterica serovar Weltevreden (BM 1643) toxoid can protect chicks against salmonellosis during their early days of life. PMID- 15782818 TI - Inhibitory effect of sildenafil on gastrointestinal smooth muscle: role of NO cGMP transduction pathway. AB - Nitric oxide (NO) is an important neurotransmitter in the gut and has been demonstrated to be a key physiological mediator of non-adrenergic non-cholinergic (NANC) relaxation of gastrointestinal smooth muscle. In the present study the effect of PDE 5 inhibitor sildenafil on the gastrointestinal function (gastric emptying and intestinal transit) has been demonstrated in mice. Sildenafil (0.5-2 mg/kg, po) did not alter the percent gastric emptying however, in higher doses (5, 10 and 30 mg/kg, po) it inhibited the gastric emptying. On acute administration (0.5-5 mg/kg, po) it did not alter the intestinal transit but in higher doses (10 and 30 mg/kg, p.o.) delayed the intestinal transit. Further, the inhibitory effect of sildenafil was significantly blocked by L-NAME (10 mg/kg, ip), a non-selective NOS inhibitor and methylene blue (1 mg/kg, ip), a guanylate cyclase inhibitor. These findings suggest the participation of NO-cGMP transduction pathway in the inhibitory effect of sildenafil (higher doses) on the gastrointestinal smooth muscles and its potential application in patients with nutcracker oesophagus, hypertensive lower oesophageal sphincter (LOS), achalsia and diabetic gastroparesis or colitis where there is a loss of nNOS. PMID- 15782819 TI - Experimental therapeutic intervention with ascorbic acid in ethanol induced testicular injuries in rats. AB - Ascorbic acid treatment significantly increased the activities of testicular delta5, 3beta-HSD and 17beta-HSD. Moreover, the treatment was also associated with significant decrease in oxidative stress in the testis. Ethanol induced oxidative stress and decreased steroidogenesis can be reversed by treatment with ascorbic acid. PMID- 15782821 TI - Antiulcerogenic effect of Justicia prostrata Gamble. AB - Antiulcerogenic effect of the alcoholic (ALJP) and aqueous (AQJP) extracts of the whole plant of Justicia prostrata was studied in aspirin+pylorus ligated rat models and analysed for gastric volume, ulcer index, free and total acidity. Biochemical parameters like total proteins, total hexoses, hexosamine, fucose and sialic acid were also estimated. Both extracts (ALJP and AQJP) significantly reduced both the gastric volume and the acidity of gastric juice. It also significantly promoted gastric mucus secretion by increasing total carbohydrates and decreasing the protein concentration in aspirin+ pylorus ligated rats. The results suggest that both the extracts (ALJP and AQJP) possess antiulcer activity, whereas AQJP is more effective when compared with ALJP in aspirin+pylorus ligated rat models. The results were compared with the standard drug Rantidine, a H2 receptor antagonist. PMID- 15782820 TI - Effect of vitamin E supplementation on diabetes induced oxidative stress in experimental diabetes in rats. AB - Diabetes induced by streptozotocin (50 mg/kg body wt, i.p.) in the rats substantially increased the plasma glucose and malondialdehyde levels along with corresponding decrease in the antioxidants levels. Supplementation of vitamin E (200 mg/kg body wt., ip) for 5 weeks resulted in non-significant decrease in the blood glucose levels but plasma malondialdehyde levels were reduced to below normal levels. Plasma vitamin E, vitamin C, uric acid and red blood cell glutathione levels were also restored to near normal levels on vitamin E supplementation to diabetic rats as compared to control (diabetic) rats. The activities of antioxidant enzymes, catalase (EC 1.11.1.6), glutathione peroxidase (GSHPx EC 1.11.1.9), and glutathione reductase (GR EC 1.6.4.2) were also concomitantly restored to near normal levels by vitamin E supplementation to diabetic rats. The results clearly demonstrated that vitamin E supplementation augments the antioxidant defense mechanism in diabetes and provides evidence that vitamin E may have a therapeutic role in free radical mediated diseases. PMID- 15782822 TI - Effect of alliums on total lipid in plasma and tissues of Japanese quails (Coturnix coturnix japonica) with respect to age and sex. AB - Dietary onion and garlic caused an increase in the level of liver and plasma total lipids in Coturnix coturnix japonica. This increase could be due to the effect of an increased feed intake, bile production, digestion and absorption that in turn caused an increased utilisation of dietary fat, increased transfer of dietary lipids to the liver and/or due to increased lipogenesis as such. However, there was no increase in the muscle lipid content. This effect in the muscle could be due to inhibitory effect of onion or garlic on lipoprotein lipase activity. Further, the changes in the tissue total lipid level in the control group due to change in age and sex were also observed. PMID- 15782823 TI - Early and transient induction of nitric oxide (NO) in infectious bursal disease virus infection is T-cell dependent: a study in cyclosporin-A treated chicken model. AB - The level of nitric oxide (NO) in the supernatants of mitogen (PHA) stimulated lymphocyte cultures from infectious bursal disease (IBD) virus infected T-cell suppressed and immune competent chickens was monitored. The immune competent chickens when infected with IBD virus showed 4-6 folds increased levels of NO as compared to uninfected chickens. The levels of NO in T-cell suppressed chickens were comparable to uninfected control chickens, in spite of markedly increased hemorrhage suggesting that the muscular hemorrhage observed in IBD in not solely and directly related with NO production. The immune suppressed chickens that did not induce NO production after IBD virus infection showed more severe lesions and supported enhanced virus replication. Taken together it may be suggested that NO production after IBD virus infection, may exert antiviral effect since the immune suppressed chickens that failed to induce NO showed more severe disease and higher magnitude of virus replication, but does not seem to correlate with the hemorrhagic lesions which in fact may be as a result of the net outcome of various host-factors and the determinants responsible for virus virulence and virus clearance. PMID- 15782824 TI - Isolation of poly (A)+ mRNA for downstream reactions from some medicinal and aromatic plants. AB - In the present protocol for extraction of RNA, hexadecyltrimethylammoniumbromide (CTAB) and insoluble polyvinylpyrrolidone were used followed by LiCl precipitation, CsCl ultracentrifugation and finally poly (A)+ mRNA was isolated with the help of oligo(dT)-cellulose columns. The isolated poly (A)+ mRNA was found to be suitable for cDNA-AFLP and suppression subtractive hybridization applications. It is a modified and consolidated protocol based on previously described methods for isolated steps and works better for medicinal and aromatic plants. High yield of poly (A)+ mRNA coupled with its amenability for downstream reactions like RT-PCR, northern blotting and cDNA synthesis for library construction is a key feature of the present protocol. PMID- 15782825 TI - Parents behaving badly. PMID- 15782826 TI - Ultrasonography in a forward-deployed military hospital. AB - Military medical units must be flexible and mobile to keep up with maneuver units on the modern battlefield. The requirements for unit mobility and maneuverability preclude bulky advanced radiologic imaging support, such as computed tomography or magnetic resonance imaging systems. Portable sonography is rapid, reliable, efficient, and user-friendly; it markedly expanded the diagnostic capability of our forward-deployed combat support hospital during Operation Iraqi Freedom. More than 400 ultrasound studies were performed during the first 6 months of hospital operations in Iraq. The use of this technology on the battlefield improved our ability to provide definitive, quality, medical care far forward and to preserve the fighting strength of supported units. PMID- 15782827 TI - Air Force expeditionary medical support unit at the Houston floods: use of a military model in civilian disaster response. AB - From June 6 to 10, 2001, Tropical Storm Allison delivered 40 inches of rain to the city of Houston, Texas. Nine of the city's hospitals were closed or severely curtailed services as a result of the flooding. All area hospitals were full to capacity, intensive care unit beds were unavailable, and patient wait times for emergency department care were 18 to 21 hours. Emergency department and intensive care unit congestion placed the entire emergency medical system of Houston in jeopardy. In response to a Federal Emergency Management Agency request, the Air Force deployed a 25-bed expeditionary medical support field hospital to Houston on June 13, 2001. The expeditionary medical support unit treated its first patient only 3.5 hours after arrival and was fully operational 8 hours later. During its 11-day stay, the facility treated 1,036 patients, including 312 ambulance arrivals, 48 inpatients, and 33 intensive care unit patients, and performed 33 dental procedures and 16 operations. PMID- 15782828 TI - Developing a pharmaceutical formulary for joint military medical operations. AB - This article describes the development of a standardized formulary for medical contingency operations for any theater of operations. The article compares peacetime health care systems within the Department of Defense and the formulary systems developed and used within the fixed facility environment with Department of Defense contingency health care operations systems and the complications encountered while attempting formulary development for the deployed environment. Despite great difficulties, the Joint Readiness Clinical Advisory Board developed, published, and marketed a jointly approved, standardized, modernized formulary to assist forces deploying for Operations Enduring Freedom and Iraqi Freedom. The process used by the Joint Readiness Clinical Advisory Board during formulary development is described in detail, giving readers an understanding of the foundations of the Joint Deployment Formulary. The military departments will experience the benefits of enhanced supply chain predictability and responsiveness, increased clinician satisfaction, and improved patient safety and health care quality by implementing the Joint Deployment Formulary for their forces engaged in contingency operations. PMID- 15782829 TI - Unprotected percutaneous coronary intervention is safe and effective for treating unstable angina in the modern warrior: the second Gulf War experience at Landstuhl Regional Medical Center, Germany. AB - BACKGROUND: Evacuation of soldiers with acute coronary syndromes to U.S. hospitals has been the treatment strategy of choice. However, percutaneous coronary intervention (PCI) complication rates have decreased to the point that it is feasible to bring this therapy forward. METHODS: Patients presenting with acute coronary syndromes during Operation Iraqi Freedom (March 2003 to June 2003] were evacuated to Landstuhl Regional Medical Center, Germany. Patients with a high clinical risk but a low interventional risk were selected for unprotected PCI. RESULTS: PCI was successfully accomplished for 93% of patients (13 of 14 patients). One patient had a 1-month history of progressive angina and a chronic total lesion that could not be crossed. There were no major complications. CONCLUSION: Patients with acute coronary syndromes who are risk-stratified can be safely treated with PCI at overseas military hospitals lacking cardiothoracic surgical back-up assistance. This strategy minimizes the risk of transoceanic evacuation of this patient population. PMID- 15782830 TI - Osteoma of the mandibular condyle: report of a case with 5-year follow-up. AB - Osteomas of the mandibular condyle are rare. An unusual case of an osteoma occurring in the mandibular condyle of a 22-year-old man with mandibular deviation and malocclusion is reported; this represents the 14th documented case in the English language literature. The tumor was resected with condylectomy. Postoperatively, mandibular deviation was minimized. PMID- 15782831 TI - Financial management using a computerized system for evaluating health care invoices. AB - The Medical Corps of the Israel Defense Forces (IDF) provides health care services for hundreds of thousands of soldiers in IDF clinics and by purchasing services from civilian institutes. Monthly invoices from civilian institutes are so numerous that most are paid with insufficient scrutiny and valuable information regarding soldiers' health care is lost. Our objective was to develop a computerized system for reviewing invoices and gathering data. Based on Oracle software (Oracle, Redwood Shores, California), the system stores the terms of agreements with medical institutes, enters billing data, calculates invoice totals, manages information, and generates reports. It automatically checks for duplicate invoices and confirms payment. The system allows users to view data for decision-making, creates insurance claim files, identifies incorrect charges, assists in quality assurance, and maintains personal patient records. With the system in operation since 2001, savings significantly increased, to approximately 5% of the IDF health care budget. On the basis of information gathered by the system, changes in medical procedures were implemented that are expected to generate even greater savings. PMID- 15782832 TI - Post-traumatic stress disorder symptoms among military health professionals in Turkey. AB - This descriptive study investigated traumatic experiences and post-traumatic stress disorder symptoms among military professionals in Turkey. The sample included 225 military health professionals who were in charge in districts in which traumatic events occurred frequently in the past decade. The Impact of Event Scale, Beck Anxiety Inventory, and Symptom Checklist were used to obtain data in the study. More than one-half of the health professionals had experienced at least one traumatic event. Those who had a history of traumatic experience and loss of relatives or friends reported more symptoms than did those who had not experienced traumatic events. As expected, health professionals who had traumatic experiences were more likely to have higher Impact of Event Scale scores than were those who had not experienced physical or psychological trauma. The findings indicate that health professionals are as affected by traumatic events as are those in other settings or occupations. Future research should focus on the long term effects of post-traumatic stress among health professionals. PMID- 15782833 TI - Failure of fenoldopam to control severe hypertension secondary to renal graft rejection in a pediatric patient. AB - We report the first case of the failure of fenoldopam for a pediatric patient with severe hypertension and renal failure. Our patient was a 3-year-old child with severe hypertension secondary to renal graft rejection and high-dose steroid treatment. The patient's hypertension was controlled with nitroprusside but not with fenoldopam. PMID- 15782834 TI - Folliculitis after smallpox vaccination: a report of two cases. AB - Rashes are frequent and potentially serious adverse consequences of smallpox vaccination. Life-threatening rashes must be differentiated from benign, self limiting ones. Generalized vaccinia, erythema multiforme, and folliculitis are distinct self-limiting entities but may be difficult to differentiate from one another. Two cases of folliculitis after smallpox vaccination are described. Both patients received anthrax vaccination within 2 weeks before smallpox vaccination. Both presented with a papulopustular rash 9 days after smallpox vaccination. Although the rashes were initially diagnosed as erythema multiforme, the clinical features were more consistent with folliculitis. Self-limiting rashes after smallpox vaccination are common and may be difficult to distinguish from each other. These rashes are clinically distinct, with characteristic features. Improvement in diagnosis may help classify the frequency and risk of rashes after smallpox vaccination. The association of vaccine-associated folliculitis, anthrax vaccine, and other potential antigenic triggers should be further explored. PMID- 15782835 TI - Mitigating effect of Department of Veterans Affairs disability benefits for post traumatic stress disorder on low income. AB - OBJECTIVE: The goal was to assess the impact of Veterans Affairs (VA) disability benefits for post-traumatic stress disorder (PTSD) on veterans' odds of poverty. Women and African American veterans were of special interest, because they are less likely than other groups to receive PTSD disability benefits. METHODS: A cross-sectional survey of 4,918 veterans who applied for VA disability benefits for PTSD between 1994 and 1998 was performed. Responses were linked to administrative data. RESULTS: Overall, 42% reported low income (defined as household income less than or equal to 20,000 dollars per year). Men's and women's odds of reporting poverty were similar, but receipt of PTSD disability benefits mediated African American veterans' odds of poverty. Veterans' odds of impoverishment were reduced considerably if they received VA PTSD disability benefits and identified themselves as disabled. CONCLUSIONS: VA disability benefits for PTSD reduced odds of impoverishment for psychiatrically ill veterans. This effect appeared to be especially important for African American veterans and for veterans self-identifying as disabled. PMID- 15782836 TI - Status of trauma care in U.S. Army hospitals. AB - OBJECTIVE: This study documents the recent trends and current state of inpatient trauma care in U.S. Army hospitals. METHODS: Inpatient trauma cases from Army hospitals worldwide from October 1988 through April 2001 were analyzed. Facilities included 3 Certified Trauma Centers (CTCs), 7 non-CTC Army Medical Centers, and 42 Army Community Hospitals. Logistic regression identified mortality risk factors. RESULTS: Overall, the Army treated 166,124 trauma cases, with a mortality rate of 0.8% (trend of 0.66% to 1.18% in fiscal years 1989-2000, p < 0.0001). The number of Army hospitals decreased by 44% and the number of trauma cases decreased by nearly 75%. Injury severity, patient age, hospital trauma volume, beneficiary category, hospital type, and a resource intensity measure were all significantly associated with the probability of death. CONCLUSIONS: The overall trauma mortality rate at Army hospitals during the study period was lower than that reported for civilian trauma centers. However, changes in patient profiles, increased average severity, and decreased trauma volume might have contributed to a 13% increase in mortality rates at CTCs. PMID- 15782837 TI - Is testicular cancer related to Gulf War deployment? Evidence from a pilot population-based study of Gulf War era veterans and cancer registries. AB - The possible relationship between military deployment and the subsequent increase in cancer rates has been prominent since the Vietnam War. The objective of this study was to investigate whether any form of cancer was increased among veterans deployed to the Persian Gulf in the 1991 conflict. This study matched data from central cancer registries in the District of Columbia and New Jersey with the records for 1.4 million Gulf War era veterans, i.e., 621,902 veterans who arrived in the Persian Gulf before March 1, 1991, and 746,248 non-Gulf veterans. Using a proportional incidence ratio, testicular cancer was found to be the only significantly increased malignancy among deployed Persian Gulf War veterans. The increase became apparent 2 to 3 years after the Persian Gulf War and peaked 4 to 5 years afterward. Our data and those of investigators studying Vietnam veterans suggest that testicular cancer may be related to military deployment. PMID- 15782838 TI - Primary care providers' attitudes toward prostate cancer risk factors at a Veterans Affairs health care facility. AB - OBJECTIVE: We asked what factors influence primary care providers' decision to screen patients for prostate cancer. METHODS: A survey completed by 175 Veterans Affairs primary care providers queried whether patient anxiety, family history, race, and other assorted risk factors increased their likelihood of screening for prostate cancer. Subsequent questions assessed the degree to which various factors, such as age, comorbidities, and lack of interest, decreased their likelihood of screening. RESULTS: The African American race increased the tendency for screening for 84.6%, followed by a family history of prostate cancer for 73.3%. Life expectancy of less than 5 years substantially decreased the tendency to screen for only 42.3%. Only 28% thought that age of more than 75 years was a deterrent to screening. CONCLUSIONS: Veterans Affairs primary care providers recognize the need to aggressively screen African Americans and men with a family history of prostate cancer. However, they often screen men with a limited life expectancy or advanced age. PMID- 15782839 TI - Detection of depression in primary care. AB - OBJECTIVE: The first purpose of this study was to examine the variables related to the diagnosis of a depressive disorder and associated with referral to a mental health clinician. The second purpose was to examine the level of agreement of the primary care provider's diagnosis of depression with that of the mental health clinician. METHODS: The design was a retrospective, case-control study (n = 162). Data were analyzed with descriptive statistics and logistic regression. RESULTS: Referred patients tended to be Caucasian, middle-aged or older men who had a history of depression, were experiencing depressive symptoms and/or an acute or chronic stressor, had a diagnosis of cardiovascular disease or cerebrovascular accident, and were taking a greater number of medications. Mental health clinicians agreed with the primary care clinicians' diagnoses of depression 76% of the time. CONCLUSIONS: Primary care clinicians accurately diagnosed depression for the majority of patients and appropriately referred them for psychiatric care. PMID- 15782840 TI - Elbow arthrodesis after war injuries. AB - Arthrodesis is a surgical procedure that results in fusion of the joint and bony ankylosis. With this operation, we can achieve satisfactory function of the limb in cases in which options for different treatments no longer exist. Severe joint destruction, joint infections, and nonunions, as well as complex war injuries (with large bone and soft tissue defects), are indications for arthrodesis. The elbow is the most susceptible joint of the upper limb to war injury. Between 1992 and 1995, we performed elbow arthrodesis for nine patients in our department after war injuries. As analysis of our results shows, elbow arthrodesis, although a rarely performed surgical procedure, is the best treatment for patients with complex war injuries of the elbow. For arthrodesis, we used external fixation in combination with internal fixation (cancellous bone screw) and additional autologous cancellous bone grafts. PMID- 15782841 TI - Treatment of temporomandibular joint ankylosis with temporalis superficial fascia flap. AB - Temporomandibular joint (TMJ) ankylosis is characterized by the formation of bony or fibrous mass, which replaces the normal articulation and limitation of mouth opening. This study aims to determine the efficacy of arthroplasty and interpositional fascia flap in the treatment of unilateral and bilateral TMJ ankylosis in three young adult men. Our operative protocol for unilateral and bilateral TMJ ankylosis entailed resection of ankylotic mass, intraoral ipsilateral and bilateral arthroplasty, interpositional tissue transfer to the TMJ with temporalis superficial fascia flap, maxillomandibular fixation, and early mobilization and aggressive physiotherapy. Early postoperative initial exercise, physiotherapy, and strict follow-up play an important role in preventing postoperative adhesions. The temporalis superficial facia flap is an autogenous graft that has the advantages of close proximity to the TMJ minimal surgical morbidity, and successful clinical results. It was found to be a valuable option for TMJ ankylosis reconstruction. PMID- 15782842 TI - Career History Archival Medical and Personnel System. AB - The Career History Archival Medical and Personnel System is a database that provides information on cancer, chronic diseases, occupational and preventive medicine, epidemiological research, and the use of health care in the Navy and Marine Corps. It was created at the Naval Health Research Center for enlisted Navy personnel, and it is being expanded to encompass all military personnel. Its objective is to provide a comprehensive, chronologically ordered database of career and medical events in all active duty military service members and to track career and disease events in order from the date of entry to service to the date service ended. Events include the dates of beginning and ending of each specific military occupation, all assignments to a military units or ships, all hospitalized diseases, and other events. The database contains detailed epidemiological data on more than six million members of the military services. It is the largest known epidemiological database in the United States. PMID- 15782843 TI - Just what can we expect from new head of HHS? PMID- 15782844 TI - Pediatric groups' revenue increase falls behind boost in practice costs. PMID- 15782845 TI - Power to the people: Newt's battle cry. PMID- 15782847 TI - Managed care (and everyone else) unprepared for the next killer flu. PMID- 15782846 TI - Information: it's better when you share. PMID- 15782848 TI - Managing drugs for rare genetic diseases: trends and insights. AB - Managed care organizations generally pay for expensive drugs that treat rare genetic diseases because few patients have these conditions, the conditions are often life threatening, and the benefit design mandates coverage. In most cases, the cost-control measures and management restrictions applied to many other specialty pharmaceuticals do not make sense for the orphan drugs used to treat extremely rare genetic conditions because treatment alternatives usually are lacking for rare conditions (e.g., Gaucher's disease) and because the per member per month costs for truly rare conditions are low. The increasing number of biologic and injectable therapies for more common conditions, however, is prompting managed care decision makers to manage these new therapies more actively. There is a potential that treatments for rare disorders will be swept up in this broad-based response. This article provides definitions, background, and stakeholder perspectives on this topic and describes recent trends and challenges in health system management of exceedingly rare genetic diseases. The author suggests that current protocols are appropriate for managing rare and ultra-rare diseases, and that applying more active management practices to less rare diseases is neither efficient nor productive; in fact, it may be counter productive. PMID- 15782849 TI - Biggest increases in specialty drug costs seen in rheumatologic, cancer, and hemophilia agents. PMID- 15782850 TI - Big companies push for better online tools. PMID- 15782851 TI - Competition heralds beginning of bio-generics. PMID- 15782852 TI - Medicare drives plan membership; Wall Street moderately bullish. PMID- 15782853 TI - [IOP in rabbits after instillation of a mixture of the antiglaucomatic Cosopt and the aminoacid L-argininin HCL into the conjunctival sac]. AB - OBJECTIVE: The article documents results of experiments focused to influence the physiological IOP values after application of the COSOPT eye drops (double combination of bbb-blocker and carbonic anhydrase inhibitor), aminoacid 10% L arginin.HCl and their mixture in rabbits. METHODS: COSOPT followed by 10% L arginin.HCl and mixture of 10% L-arginin.HCl with COSOPT was instilled weekly into the left conjunctival sac of 5 adult rabbits (females of the New Zealand White species). The IOP and pupilar diameter were measured in 5, 15, 30, 60, 120, 180, 240 min. and 24 hours after instillation. The right eye was used as control. RESULTS: The antiglaucomatic COSOPT no significantly decreased the rabbit IOP during 24 hours (mean value 1.85 %) in vivo. The aminoacid 10% L-arginin.HCl achieved significant decrease of the IOP: the mean value was 16.03% reaching maximum in 60th and 180th min. After instillation of the 10% L-arginin.HCl and COSOPT mixture in the IOP showed significant decrease with mean value of 9,64% (except values in 5th min. and 24th hour). Compared with control eye, the major decrease of the IOP was measured in 240th min. (mean value 17.98%). However even in 24 hours after instillation the IOP was not reaching the values of the control eye, shoving also the IOP decrease. CONCLUSION: We assume that in a double combination of antiglaucomatics in COSOPT a new substance was created. This substance interacted in vitro with the aminoacid 10% L-arginin.HCl resulting in a new "bio-antiglaucomatis" better penetrating into the target area. This new substance was responsible for more significant decrease after the mixture application compared with COSOPT alone. The presence of aminoacid can slow down the undesirable effect of the antiglaucomatics together with the possible neuroprotection. PMID- 15782854 TI - [Effect of viscoelastic substances on postoperative intraocular pressure in phacoemulsification]. AB - The Influence of the Type of Viscoelastic Substances on the Level of Intraocular Pressure after Phacoemulsification The aim of study was to asses the influence of the type of viscoelastic substances on the level of intraocular pressure (IOP) after cataract surgery and intraocular lens (IOL) implantation. In the study group of 100 patients Viscoat was used at 32 operated eyes, Provisc at 14 eyes and Duovisc in 54 eyes during phacoemulsification and IOL implantation. IOP was measured before surgery, one day after surgery and one week after surgery. Mean IOP level 1. day postoperatively was 24.94 mmHg in the Viscoat group of eyes, 24.65 mmHg in the Provisc group and 21.09 mmHg in the Duovisc group. The type of viscoelastic substance used during the surgery significantly influenced the level of IOP in the first postoperative day. The level of IOP was significantly lower in the Duovisc group comparing to Viscoat and Provisc. The level of IOP did not differ significantly between the Viscoat and Provisc group. One week after surgery the level of IOP was not significantly different between all types of viscoelastic device. Duovisc was the most profitable viscoleastic substance in the point of view of the course of phacoemulsification, IOL implantation and postoperative level of IOP. PMID- 15782855 TI - [Early functional effect of the pars plana vitrectomy in complications of the proliferative diabetic retinopathy]. AB - During the 1997-2002 period, 48 eyes of 41 patients of the mean age 58 years were operated on because of complications of proliferative diabetic retinopathy. In the cohort of operated patients, the type I diabetes mellitus was determined in 10 (21%) cases, the type II diabetes mellitus in 36 (75%) cases, and in two cases (4%), another type of diabetes was detected. One patient of those two had LADA type diabetes and the second one had secondary diabetes due to chronic pancreatitis. In the early postoperative period, or during first three months postoperatively, after the pars plana vitrectomy, the visual acuity (VA) in 28 (59%) eyes improved, in 16 (33%) eyes remained the same, and in 4 eyes (8%) worsened. VA 1/60 (0.017 or 3/200) and better had 37 (77%) eyes, VA 6/60 (0.1 or 20/200) and better had 17 (37%) eyes, and VA 6/12 and better (0.5 or 20/40) had 3 (6%) eyes only. VA worse than 1/60 (0.017 or 3/200) had 11 (23%) eyes. Authors emphasize the importance of regular and detailed ophthalmologic examinations with early diagnosis of pathological changes and early start of adequate treatment. PMID- 15782856 TI - [The cases of penetrating eye injuries with large intraocular foreign body]. AB - AIM: Three consecutive cases of penetrating eye injuries complicated with large (size 9-12 mm) intraocular foreign body are presented as such. PATIENTS AND METHODS: The initial findings, management, surgical procedures and final anatomical and functional outcomes of each particular case are given. Corneal entrance laceration was seen in all three patients. Second scleral full-thickness eye wall laceration was found just posterior to the horizontal muscle insertions in two eyes. Full-thickness exit posterior eye wall laceration obturated with foreign body was diagnosed in one case. This metallic foreign body projected into posterior orbit. Two eyes were injured with metallic radio opaque foreign body. Glass fragment was removed from posterior segment of the eye in one patient. Following primary wound closure pars plana vitrectomy was performed to remove all posterior segment intraocular foreign bodies. RESULTS: Silicone oil was used to fill the vitreous cavity at the end of the surgery in two eyes. Gas bubble (perfluoropropane, C3F8) was injected into the vitreous space at the end of the vitrectomy in one eye. The authors observed following complications: vitreous cavity haemorrhage, traumatic cataract formation, cystoid macular oedema and peripheral stationary traction retinal detachment. CONCLUSION: Good anatomical results and restoration of good visual functions of injured eye were achieved in all patients despite some inferior initial prognostic factors. PMID- 15782857 TI - [Ligneous conjunctivitis: complication of inborn plasminogen deficiency (a case report)]. AB - Authors refer about detailed analysis of ten years follow up of a child patient with ligneous conjunctivitis. They document presence of all typical eye findings (especially recurrent formation of granulomatous pseudotumors in affected mucous membrane) related to this rare pseudomembranous conjunctivitis. The clinical picture of the disease includes plasminogen deficiency, a factor newly considered as primary cause of the disease. The diagnosis was confirmed histologicaly by repeated probatory excisions. Pseudomembranous inflammation with mixed inflammatory infiltrate containing large amount of elements of chronical as well as acute inflammatory reaction is typical. Granuloma with the accumulation of the PAS-positive amorphous matrix with high content of the fibrin and with the network of newly formed vessels is the component of the inflammatory picture. The treatment was based on successively discovered knowledge of the possible etiopathogenesis of the disease. Only the ninth, last surgical procedure induced longer-term remission, which lasts sixteen months until now. The granuloma excision was combined with the mitomicin application on the wound surface and a conjunctivoplasty. During the postoperative period, the ointment with heparin, corticosteroid and antibiotic was applied. The purpose of the mitomicin use is to slow down the fibroproliferative reparative reaction. Covering of the wound surface decreases the number of microtraumas on the uneven postoperative scleral surface. Heparin in the interstitial tissue of the wound surface blocks the conversion of fibrinogen to fibrin. PMID- 15782858 TI - [Evaluation of visual function and changed quality of life among patients after excimer refractive surgery]. AB - An anonymous wide-ranging questionnaire was forwarded to 60 patients (40 women and 20 men) who had a laser refractive surgery for myopia. The questionnaire was focused on 1. reasons for seeking a refractive surgery, 2. subjective complaints shortly after a surgery, 3. postoperative status: functionality, 4. satisfaction. The satisfaction with final outcome has reached 93% and that closely correlates to results of uncorrected VA (UCVA). However, some patients (17%) reported reduced vision under dim illumination and at night and also difficulty with nighttime driving. PMID- 15782859 TI - [Ultrastructure of the iris pigmentary epithelium]. AB - The iris pigmentary epithelium has identical structure as the retinal epithelium has. The peripheral iridectomy makes it possible to obtain sufficient amount of tissue to be used for the multiplication of the pigmentary epithelium cells in tissue culture, or, for direct implantation into the subretinal space. The technique of the peripheral iridectomy is regardful enough; in none of the specimens mechanically damaged cells were found. PMID- 15782860 TI - [Pigmentary glaucoma after implantation of the iris claw intraocular lens]. AB - The authors refer about a female patient with pigmentary glaucoma after implantation of iris claw intraocular negative dioptric power lens. Because of high myopia, eight years ago, a phakic iris claw intraocular negative dioptric power lens was implanted in the right eye and a photorefractive keratectomy (PRK) was performed in the left eye in a 32 years old woman. During the postoperative examinations the patient complained about headaches, and in the right eye elevated intraocular pressure was detected. On the intraocular lens surface, as well as in the anterior chamber angle, a dispersion of pigmented cells was detected and the perimetric examination revealed visual field defects characteristic for glaucoma. Findings in the left eye were normal. In the literature, reports about secondary glaucoma in eye with implanted iris claw lens and functional iridectomy are not so common. With growing number of implanted phakic iris claw intraocular lenses, the long-term follow up of those patients will be useful. PMID- 15782861 TI - [Vision quality in premature children]. PMID- 15782862 TI - [Jacques Daviel, 11 August 1696--30 September 1762]. PMID- 15782863 TI - [Report of 2 new types of eye surgery diametrically different from each other]. PMID- 15782864 TI - Causes of death classified by risk and condition, New Zealand 1997. AB - OBJECTIVE: To classify causes of death in New Zealand by risk factor (in addition to condition) as a planning tool for health promotion. METHOD: Deaths occurring in New Zealand in 1997 were classified by 20 prevalent risk factors using a combination of categorical attribution (rule-based) and counterfactual modelling (population-attributable risk-based) approaches. RESULTS: Approximately 30% of deaths were attributed to the joint effect of dietary factors. Tobacco consumption was responsible for 18% of deaths and insufficient physical activity for almost 10%. Less important behavioural risk factors included alcohol consumption (3%), illicit drug use (0.5%) and unsafe sex (0.5%). Among biological risk factors, higher than optimal total blood cholesterol, systolic blood pressure and body mass index accounted for 17%, 15% and 12% of deaths respectively. Deprivation contributed to 17% of deaths, and adverse in-hospital events to 6%. Among environmental exposures, microbes accounted for 6.5% of deaths, air pollution 3.5% and occupational diseases and injuries 0.5%. Among injury hazards, risk factors related to road traffic were responsible for 2% of deaths, while violence accounted for 2.5% of deaths, mostly through suicide. Cross-classifying deaths by both condition and risk factor, 90% of ischaemic heart disease and 80% of stroke, but only 30% of cancer deaths, could be attributed to specific risk factors. CONCLUSIONS: This is the first comprehensive ranking of causes of death at the level of risk factors available for New Zealand and should prove useful as a planning tool, especially for disease prevention and health promotion. PMID- 15782865 TI - Weapon carrying in a sample of high school students in New Zealand. AB - OBJECTIVE: To examine self-report of carrying weapons and correlates in a sample of New Zealand high school students. METHOD: A sample of 652 Year 11 students from all Dunedin high schools completed a web-based version of the US Youth Risk Behaviour Survey. This included questions on weapon carrying, fighting, and feeling unsafe at school, as well as measures of family and peer/friend connectedness and perception of school climate. RESULTS: Reports of ever carrying a weapon and carrying one in the last 30 days were relatively common, being 27.5% and 19.3% respectively. Weapon carrying in the last 30 days was strongly associated with being male, fighting in the last year, missing school due to feeling unsafe, and a poorer perception of school climate. CONCLUSIONS: Many high school students report weapon carrying, which in turn was associated with other aspects of physical aggression. Further study of the lethality of weapons being carried and the context in which they might be used is clearly warranted. PMID- 15782866 TI - Sleep, sleepiness and motor vehicle accidents: a national survey. AB - OBJECTIVE: To assess the role of sleep-related factors, ethnicity and socioeconomic deprivation in self-reported motor vehicle accidents while driving, after controlling for gender, age and driving exposure. METHODS: Mail survey to a random electoral roll sample of 10,000 people aged 30-60 years, stratified by age decades and ethnicity (71% response rate). The analytical sample included 5,534 current drivers (21.6% Maori men, 21.2% Maori women, 30% non-Maori men, 27.2% non Maori women). RESULTS: Multiple logistic regression analyses revealed the following independent risk factors for accident involvement while driving (last three years): being younger; higher average weekly driving hours; never/rarely getting enough sleep (OR=1.26, 95% CI 1.06-1.49); reporting any chance of dozing in a car while stopped in traffic (Epworth Sleepiness Scale question 8, OR=1.52, 95% CI 1.15-2.02); and among women, being non-Maori. Total Epworth score was not significantly related to reported accident involvement. CONCLUSIONS: Chronic sleep restriction, and any likelihood of dozing off at the wheel of a motor vehicle, were significant independent predictors of self-reported involvement in all types of motor vehicle accidents, not only those identified as fatigue related. The Epworth Sleepiness Scale alone is not a reliable clinical tool for identifying individuals at higher risk of crashes. IMPLICATIONS: Factors relating to chronic sleepiness were as important as established demographic risk factors for self-reported motor vehicle accident involvement among 30-60 year-old drivers. The findings reinforce the need for multi-faceted campaigns to reduce sleepy driving. PMID- 15782867 TI - Prevalence and consequences of insomnia in New Zealand: disparities between Maori and non-Maori. AB - OBJECTIVE: To investigate the prevalence of self-reported insomnia symptoms among Maori (Indigenous people) and non-Maori adults in the general population of New Zealand. To explore the consequences for health and quality of life experienced by those who report common insomnia complaints and sleeping problems. METHODS: In 2001, a two-page questionnaire was mailed to a stratified random sample of 4,000 adults aged 20-59 years nationwide. Participants were selected from the New Zealand electoral roll. The sample design aimed for equal numbers of Maori and non-Maori participants, men and women, and participants in each decade of age (72.5% response rate). RESULTS: Population prevalence estimates indicate that self-reported insomnia symptoms and sleeping problems are higher among Maori than non-Maori. Multiple logistic regression analyses showed that self-reported insomnia symptoms and/or sleeping problems are significantly associated with reporting poor or fair health and quality of life outcomes. CONCLUSIONS: Approximately one-quarter of adults in New Zealand may suffer from a chronic sleep problem, highlighting insomnia as a major public health issue in New Zealand. IMPLICATIONS: Significant differences in the prevalence of insomnia symptoms and current sleeping problems with respect to ethnicity have implications in the purchase and development of treatment services, with greater need for these services among Maori than non-Maori. PMID- 15782868 TI - The epidemiology of home injuries to children under five years in New Zealand. AB - OBJECTIVE: This paper describes the epidemiology of injuries sustained by children under five in the home. METHODS: Cases were selected from the New Zealand Health Information Service public hospital morbidity and mortality data, and included all 0-4 year olds where the place of injury occurrence was classified as 'home'. The circumstances of injury were coded according to the Supplementary Classifications of External Causes of Injury and Poisoning (E codes) of the International Classifications of Diseases. Age-specific rates of death or hospitalisation due to injury were calculated using the population of 0 4 year olds in New Zealand for each year as the denominator. RESULTS: The rate of death from an injury sustained at home between 1989 and 1998 was 13 per 100,000 population per year. The main causes of death were suffocation, submersion, homicide and fire. The rate of hospitalisation in children aged 0-4 years from an injury sustained in the home between 1989 and 2000 was 737 per 100,000 population per year. The most frequently recorded causes of hospitalisation were falls, scalds, poisonings and cut/piercing incidents. CONCLUSION AND IMPLICATIONS: Although there has been an apparent decrease in the number of children hospitalised for injuries sustained in the home environment, it is not possible to determine if this is a 'real' change or a result of other factors affecting the data. While children continue to be killed and injured as a result of preventable incidents in the home environment, injury prevention strategies should be continued and strengthened. PMID- 15782869 TI - Prevalence and determinants of influenza vaccination in Australians aged 40 years and over--a national survey. AB - OBJECTIVES: To determine influenza vaccination coverage in 2001 in Australian adults aged > or = 40 years, assess awareness of and attitudes to influenza vaccine, factors associated with vaccination, and estimate uptake of free vaccine provided to those aged > or = 65 years. METHODS: National computer-assisted telephone interview (CATI) survey in October/November 2001. RESULTS: Interviews were completed with 5,266 people aged > or = 65 and 2,415 aged 40-64 years. Thirty per cent of selected households participated. Overall, 67% of respondents believed that the vaccine was somewhat to very effective in preventing influenza. Seventy-eight per cent of those aged > or = 65 years reported influenza vaccination; 89% had received it free. Independent predictors of vaccination were: belief that influenza vaccine is effective in preventing influenza (OR=13.5, 95% CI 10.6-17.2); and the presence of chronic disease (OR=1.6, 95% CI 1.3-2.0). Overall, 24% of those aged 40-64 years were vaccinated; only 34% of those who met any of the criteria for vaccination (medical risk factor, at-risk occupation, or being Aboriginal or Torres Strait Islander) reported vaccination. CONCLUSIONS: Influenza vaccine coverage was high in those aged > or = 65 years, but coverage of those at-risk aged 40-64 years remained suboptimal. Immunisation against influenza was influenced more by beliefs about the vaccine's effectiveness and existing medical risk factors, rather than socio-demographic factors such as gender and income. PMID- 15782870 TI - Prevalence of influenza vaccination in South Australian aged care homes. AB - OBJECTIVE: To determine the prevalence of influenza immunisation of residents within South Australian aged care homes, reasons why vaccination is not given, the policy associated with vaccination administration and the source of the vaccination. METHOD: A clustered, multistage, area sample was used to assess the provision of the influenza vaccine to 500 randomly selected residents within 50 randomly selected aged care homes in South Australia during face-to-face interviews with the director of nursing at each of the selected facilities. The response rate was 100%. RESULTS: Overall, 88.4% (95% CI 85.2-91.0) of residents had received an influenza immunisation in 2001. The reasons why a resident did not have an influenza vaccination included refusal and allergy. Consent was generally obtained on an ad hoc basis (in 94.0% of homes) with the most frequent form of consent type being verbal (66.0%). Influenza vaccine administration was part of standing drug orders in 16.0% of cases and over half of the homes (54.0%) had a specific policy of education and encouraging both residents and staff to be vaccinated against influenza. CONCLUSION: Residents of aged care homes, a population deemed to be at high risk of contracting influenza, are receiving an adequate level of influenza coverage although improvements could be made. Addressing policy and procedural issues can maintain, and further improve, vaccination levels. IMPLICATIONS: Clarification of policies for influenza vaccination administration particularly in regard to staff could further improve rates of vaccination coverage. PMID- 15782871 TI - An assessment of the interval between booster doses of Japanese encephalitis vaccine in the Torres Strait. AB - OBJECTIVE: Japanese encephalitis (JE) emerged for the first time in the Torres Strait, north Australia, in 1995. The inactivated mouse-brain derived JE vaccine was offered to all residents of the outer Torres Strait Islands prior to the 1996 wet season. This study was undertaken to determine the appropriateness of the recommended three-year interval between booster doses of the vaccine. METHODS: JE neutralising antibody was measured in residents of Badu Island for whom 30-36 months had passed since either a previous booster or the completion of the primary immunisation series. RESULTS: Only 70 (32%) of 219 eligible individuals had protective antibodies; 50 (37%) of the adults were immune, compared with 20 (24%) of the children (odds ratio (OR) 1.93; 95% confidence interval (CI) 1.01 3.74). CONCLUSIONS: This low level of immunity suggests that there is little in the way of natural boosting from either JE or other closely related viruses. Given the apparent low level of risk of exposure to the JE virus in the Torres Strait, and the logistical complexities involved in delivering the booster doses, the current recommendation of a three-year interval is not inappropriate. IMPLICATIONS: It would be advantageous to have a JE vaccine that is not only safer but also more immunogenic, so that it might be possible to further increase the booster dose interval. PMID- 15782872 TI - National serosurvey of poliovirus immunity in Australia, 1996-99. AB - OBJECTIVE: To measure immunity to poliovirus types 1, 2 and 3 in the Australian population. METHODS: Sera were collected opportunistically from laboratories around Australia between 1996 and 1999. A representative sample by age and gender was tested for neutralising antibodies to poliovirus types 1, 2 and 3. A titre of > or = 8 was considered antibody positive and indicative of immunity. RESULTS: Of the 1,813 sera tested, 82% were antibody positive for poliovirus type 1 and 88% were positive for type 2. Immunity to type 3 poliovirus was lower overall (74%) and especially in school-aged children and young adults. For all three poliovirus types, there were more females immune than males and immunity peaked in the 2-4 years age group. The proportion of the population immune to all three types was 59%, and 3% were negative for all three types. CONCLUSIONS AND IMPLICATIONS: This is the first national serosurvey for immunity to poliovirus in Australia. Herd immunity is probably sufficient to prevent generalised outbreaks due to type 1 and 2 poliovirus, but this may not be the case for type 3. However, localised outbreaks of any poliovirus type could still occur following reintroduction unless uniformly high levels of vaccination coverage are maintained. Ongoing serosurveillance is required following the recent change back to inactivated polio vaccine. PMID- 15782873 TI - Control strategies for Q fever based on results of pre-vaccination screening in Victoria, 1988 to 2001. AB - OBJECTIVE: Data from Q fever pre-vaccination screening were analysed to determine the level of agreement between the two tests of immunity and between disease or vaccination history and immunity, trends in proportion of participants immune to Q fever, and the annual risk of infection. METHOD: Data from nearly 10,000 screening episodes between July 1988 and June 2001 on Victorian workers at high risk were assessed. RESULTS: Most participants were male (86%) and employed in Victorian abattoirs (81%). Agreement between results of tests for immunity was 'fair' (kappa=0.52). Self-reported history of vaccination or infection was a poor predictor of immunity. The proportion of positive blood and skin tests increased with years of exposure to animals/meat, but decreased over the 13-year period with shorter exposures. Hence the percentage requiring vaccination increased from 50% in 1998 to 90% in 2001. The average annual risk of infection among abattoir workers was 45.0 per 1,000 (95% CI 42.3-47.6), and 62.6 per 1,000 (95% CI 57.5 67.7) over the first 10 years of exposure. CONCLUSIONS: This is the first Australian study to estimate the annual risk of Q fever infection in abattoir workers. The study confirmed previous findings of poor agreement between screening tests and predictive value of history of vaccination/ exposure. Up to 90% of new entrants in high-risk workplaces will be susceptible to Q fever and require vaccination. IMPLICATIONS: Systematic post-marketing surveillance is needed to monitor adverse events to the vaccine, duration of protection and possible reasons for vaccine failures. PMID- 15782874 TI - Monitoring measles elimination in Victoria. AB - OBJECTIVES: To weigh the evidence from outbreak data that Victoria has achieved, and is maintaining, elimination of measles. To identify age groups that measles vaccination has not protected adequately. METHODS: Data on observed measles outbreaks in Victoria since the start of 1998 are used to estimate the reproduction number of cases, and the probability that it is maintained below unity, its threshold value for elimination. The relative susceptibility to measles is estimated as a function of age, with confidence intervals. RESULTS: Seventeen measles introductions led to secondary cases, while 22 were single-case introductions. From these, the probability that the reproduction number for cases exceeds unity is estimated to be 0.044, or less, depending on assumptions made. There is no evidence that the reproduction number increased over time. Those aged between 19 and 32 years were most susceptible, followed by those in the first and second year of life. CONCLUSIONS: The data provide strong evidence that Victoria has maintained elimination of measles over the period 1998 to mid-2003. There is scope to improve the immunisation coverage. It is not clear how much outbreak intervention is contributing to the success in achieving apparent elimination. IMPLICATIONS: To prevent importations from causing a major epidemic of measles, Victoria must maintain its immunisation coverage and outbreak control at current levels, or better. It is important to monitor the control of measles even when elimination is achieved. PMID- 15782875 TI - Knowledge about hepatitis and previous exposure to hepatitis viruses in immigrants and refugees from the Mekong Region. AB - OBJECTIVE: Infection with hepatitis B (HBV) and hepatitis C (HCV) viruses is relatively common throughout South-East Asia and chronic infection can lead to severe consequences. This study assesses knowledge about HBV and HCV and estimates the seroprevalence of markers for these viruses in immigrants from Laos and Cambodia. METHODS: Ninety-five Laotian (aged 18-82 years) and 234 Cambodian (15-92 years) immigrants participated in separate community-based surveys conducted during 1998 and 2002, respectively. Participants completed a questionnaire on health status and level of knowledge about viral hepatitis. Blood samples were collected and tested for the presence of HBV and HCV markers. RESULTS: Nine per cent of Laotian and 8% of Cambodian participants were infected with HBV. While 49% of Laotian and 64% of Cambodian participants showed evidence of previous exposure to HBV, 30% and 9%, respectively, were vulnerable to infection. The seroprevalence of antibodies to HCV was 3% in the Laotian and 8% in the Cambodian participants. Between one-fifth and one-third of the Laotians and Cambodians who had heard of HBV and HCV knew of possible transmission routes for the viruses. Most of those with HBV or HCV infection were unaware they were infected. CONCLUSIONS: These findings indicate a significant prevalence of undetected HBV and HCV infections and an urgent need for the provision of culturally relevant information about viral hepatitis in immigrants of South-East Asian origin. PMID- 15782876 TI - Reconciling population benefits and women's individual autonomy in mammographic screening: in-depth interviews to explore women's views about 'informed choice'. AB - OBJECTIVE: To explore women's reactions to 'informed choice' in mammographic screening. SETTING AND METHODS: Telephone interviews with a convenience sample of 106 women aged 45-70 years recruited from general practices in Sydney. RESULTS: Many (42%) women preferred an active role in decision-making. Respondents had low scores for 'uncertainty' and 'factors contributing to uncertainty' in response to explicit questions about the decision to have mammographic screening. Yet respondents indicated significantly greater willingness to have a test when the benefit of a 'new' screening test for breast cancer was expressed as relative risk reduction (RRR) (88%) than either absolute risk reduction (ARR) (78%) (McNemar's test: chi(2)1=7.14, p=0.013) or all-cause mortality (53%) (McNemar's test: chi(2)1=35.1, p<0.01). Significantly more respondents considered information about ARR 'new' to them (65%) compared with RRR information (30%) (McNemar's test: chi(2)1=25.83, p<0.01). CONCLUSIONS: As mammographic screening exposes well women to potential harms for an overall population benefit, it is challenging to ensure 'informed choice'. Our results suggest women will likely appreciate individual consultations as the context in which to share complex information that women in our study agreed they need to know about mammographic screening. Our results also demonstrate that women's willingness as individuals to participate in mammographic screening is influenced by 'framing effect'. Hence, the quantitative content of decision aids to promote 'informed choice' must be comprehensive and balanced. PMID- 15782877 TI - How much does a reminder letter increase cervical screening among under-screened women in NSW? AB - OBJECTIVE: To evaluate a direct mail-out campaign to increase Pap screening rates in women who have not had a test in 48 months. METHODS: Ninety thousand under screened women were randomised to be mailed a 48-month reminder letter to have a Pap test (n=60,000), or not to be mailed a letter (n=30,000). Differences in Pap test rates were assessed by Kaplan-Meier survival analysis, by chi2 tests of significance between Pap test rates in letter versus no-letter groups, and by proportional hazards regression modelling of predictors of a Pap test with letter versus no-letter as the main study variable. T-tests were conducted on mean time to Pap test to assess whether time to Pap test was significantly different between the intervention and control groups. RESULTS: After 90 days following each mail-out, Pap test rates in the letter group were significantly higher than in the non-letter group, by approximately two percentage points. After controlling for potential confounders, the hazard ratio of a Pap test within 90 days of a mail-out in the letter group was 1.5 compared with 1.0 in the no-letter group. Hazard ratios of having a Pap test within 90 days decreased significantly with time since last Pap test (p<0.0001); were significantly higher than 1.0 for most non-metropolitan areas of NSW compared with metropolitan areas; and increased significantly with age (p<0.0001). Pap test hazard ratios were not associated with socio-economic status of area of residence, but the hazard ratio was significantly higher than 1.0 if the reminder letter was sent after the Christmas/New Year break. No significant differences in mean time to Pap test were found between the letter and no-letter groups. CONCLUSIONS AND IMPLICATIONS: Being sent a reminder letter is associated with higher Pap testing rates in under screened women. PMID- 15782878 TI - Documenting end of life decisions in residential aged care facilities in South Australia. AB - OBJECTIVE: This research examined the provision of palliative care for residents with a non-cancer diagnosis including the use of advance directives and advance care planning as part of palliative care policies in residential aged care facilities in South Australia. There are no guidelines for recording residents' wishes if they are no longer competent. METHODS: Stage 1 involved a survey of 90 randomly selected aged care facilities. Stage 2 involved case studies of 69 residents, appropriate for palliative care, from 17 facilities and interviews with 15 directors of care. RESULTS: Most residential aged care facilities used forms to record residents' wishes about end of life care, but there was little consistency. Some had no palliative care policy and few facilities required a formal advance directive. Not all residents had formally appointed a proxy. CONCLUSIONS: Residential aged care facilities should be required to develop and implement a palliative care policy acknowledging the Accreditation Standards and State legislation and including a simple tool for advance care planning. Case conferencing could assist in discussing and documenting the resident's wishes. Public education is essential to increase community and professional awareness in order to promote empowerment for the increasing number of older people who will die in residential aged care facilities. PMID- 15782879 TI - Brushtail possums (Trichosurus vulpecula) may pose a threat to public health in New Zealand. PMID- 15782880 TI - Wisdom from big pharma's Dr. Gloom. Interview by Abrahm Lustgarten. PMID- 15782881 TI - How Alfred Sloan, Michael Porter, and Peter Drucker taught us all the art of management. PMID- 15782882 TI - How to battle the coming brain drain. PMID- 15782883 TI - Ichan the spoiler. PMID- 15782884 TI - Why Johnny can't save for retirement. PMID- 15782885 TI - Gender difference in prevalence of upper extremity musculoskeletal symptoms among Swedish pig farmers. AB - There have been very few epidemiological studies dealing with pig farmers' musculoskeletal health. The aim of this study was to carry out a cross-section postal questionnaire survey dealing with musculoskeletal symptoms among female and male pig farmers in large-scale production. The participation rate overall was 70% (288/410). Over 10% of the questionnaires had missing gender data. Three different questionnaires were used: the general standardized Nordic questionnaire for the analysis of musculoskeletal symptoms; a questionnaire dealing with occurrence of numbness, reduced muscle strength, etc., in the wrists and hands; and occupational factors were screened by a special questionnaire. The results showed that musculoskeletal morbidity is high among pig farmers. The women had significantly more problems than the men with respect to the upper extremities. Symptoms in the wrists and hands such as numbness, reduced muscle strength, aching fingers and wrists, and tendency to drop things were significantly more common among the women than the men. Occupational factors dealing with, for example, the size of the pig farm, were not related to the occurrence of symptoms. Occupational factors of importance for the development of disorders, particularly among women pig farmers, should be given priority in ergonomic interventions. PMID- 15782886 TI - Self-reported musculoskeletal symptoms and observed risk factors in bareroot tree nurseries. AB - A cross-sectional study of jobs at four bareroot reforestation tree nurseries in Washington and Oregon investigated the prevalence of musculoskeletal symptoms and potential work-related musculoskeletal disorders (WMSDs), and analyzed their association with physical and psychosocial risk factors of the jobs. Questionnaires were used to assess symptoms and psychosocial factors. Direct observational work sampling was utilized to estimate physical risk factors. The response rate for the questionnaires was 41% (203 subjects), of which 72% reported recurring symptoms in the past year. The most common body region was the wrist/hand (42%). Pain was the most commonly reported symptom. Forty-one percent of the subjects had at least one potential WMSD, the hand region having the greatest number of cases (n = 51). The average age of those with and without potential WMSDs did not significantly differ; however, tenure at the nursery (p < 0.03) did. Being female (OR = 7.37; 95% CI = 2.75, 19.7,) high job satisfaction (OR = 0.32; 95% CI = 0.15, 0.72), and having a second job (OR = 3.76; 95% CI = 1.12, 12.57) were significantly associated with potential WMSDs. No significant difference in WMSD prevalence was found between the field and packing shed areas of the nurseries (p = 0.88). Pinch gripping was observed 24% of the time in the shed and 8% of the time in the field. Torso flexion was observed more often in the field than the shed (38% vs. 18% of the time). This study found that both physical and psychosocial factors associated with WMSDs are present in bareroot trees nurseries. PMID- 15782887 TI - Safety-related knowledge and behavior changes in participants of farm safety day camps. AB - Significant resources are devoted to conducting farm safety day camps throughout North America, but the impact and effectiveness of these programs has not been systematically demonstrated. This project assessed changes in safety-related knowledge and behaviors among participants in the Progressive Farmer Farm Safety Day Camp program. A written pre-test and a three-month telephone post-test were administered to three samples of participants, ages 8 to 13, in camps held in 1999, 2000, and 2001. A sample of 20 to 30 camps was included in each year of the study, with a total sample of 1,780 participants for all three years. The pre test and post-test contained questions related to first aid and to safety around animals, ATVs, farm equipment, flowing grains, and tractors. Three scores were computed from responses to 20 knowledge and behavior items. A knowledge score indicated the number of 8 knowledge items answered correctly, a behavior risk score indicated the amount of risk exposure for the child based on 8 behavior items, and an ATV safety gear risk score indicated, for those who rode ATVs, the level of risk due to lack of proper safety gear (4 items). From pre-test to post test, there was an increase in knowledge scores and a decrease in behavior risk scores and ATV safety gear risk scores. These changes were consistent both for males and females, for farm residents and non-farm residents, and across all ages in the sample. These results support claims for the effectiveness of farm safety day camps for increasing knowledge and improving safe practices among camp participants. PMID- 15782888 TI - Pesticide safety training and access to field sanitation among migrant farmworker mothers from Starr County, Texas. AB - Very little published research describes employer compliance with EPA-mandated Worker Protection Standard (WPS) pesticide safety training and the OSHA Field Sanitation Standard among farmworker women in general and mothers specifically. A goal of both standards is limiting farmworkers' exposure to potentially hazardous agricultural pesticides. Data from a NIOSH-supported cohort study ("Injury and Illness Surveillance in Migrant Farmworker Families") allowed for examining these issues. The cohort included 267 migrant farmworker families who usually reside along the Texas-Mexico border (Starr County, Texas). Data were collected in Starr County during in-home interviews. Of 102 mothers who participated in migrant farm work during summer 2001, 57 (55.9%) reported having ever received training/instruction in the safe use of pesticides, while 47 (46.1%) reported having received training within the previous five years, as required by WPS. Of trained mothers, 91.5% to 93.6% reported that their training covered key WPS areas: (1) entry into a recently treated field, (2) pesticide related injuries/illnesses, and (3) where to go and who to contact for emergency care following exposure. Regarding access to field sanitation, 67.5% to 84.2% of 77 mothers who worked outside Texas reported employer-provided decontamination supplies (e.g., soap, wash water, towels, and toilet facilities). However, a strikingly smaller proportion (12% to 28%) of 25 mothers who worked within Texas reported access to the same resources, suggesting discrepancies in compliance across the U.S. Due to the low level of employer compliance with both WPS and OSHA mandated standards, increased enforcement and an alternate delivery of pesticide training is recommended. PMID- 15782889 TI - Utilizing expert panels in agricultural safety and health research. AB - This article reports on the use of expert panels by Purdue University's Agricultural Safety and Health Program staff to address three separate research questions related to: (1) developing strategies for the prevention of flowing grain engulfments, (2) evaluation of commercial lifts used by operators with restricted mobility to gain access to agricultural tractors and equipment, and (3) identifying essential competencies for youth enrolled in federally mandated tractor and machinery certification programs. Advantages and disadvantages associated with the use of expert panels are addressed, and specific examples of outcomes are presented. Recommendations for using expert panels during research projects are also included. PMID- 15782890 TI - ROPS retrofitting: measuring effectiveness of incentives and uncovering inherent barriers to success. AB - Tractor overturns are the leading cause of all tractor- and machinery-related fatalities in the agricultural industry. A rollover protective structure (ROPS) on a tractor is the most effective mechanism for protecting a tractor operator's life during a tractor overturn incident. Unfortunately, about half of all tractors presently in operation in the U.S. do not have a ROPS. Retrofitting such tractors with ROPS could result in an as much as a 99% reduction in fatalities associated with tractor overturns. The overall aim of this study was to determine the level of financial incentive required to motivate the maximum number of farmers to install ROPS on non-ROPS equipped tractors and thus affect the greatest level of change within the farming community. This was done by offering a range of subsidy levels by percentage and not by specific dollar amounts to a random sampling of New York farms. A secondary goal was to find any hidden problems associated with retrofitting. Study results showed that cost was not the only factor affecting farmers' reluctance to retrofit. A perceived and actual "hassle factor" was found to be endemic to the retrofitting process and a significant obstacle to farmers' willingness to retrofit, no matter the level of financial subsidy. PMID- 15782891 TI - Lessons learned: geographic information systems and farmworkers in the Lake States. AB - Agencies serving the estimated 42,000 to 137,000 migrant and seasonal farmworkers in the Lake States (Wisconsin, Michigan, and Minnesota) face distinct challenges, including inadequate access to farmworker data to address their needs. This project developed and evaluated a geographical information system (GIS) database for compiling and displaying existing farmworker data in the Lake States. A three step study was conducted in the Lake States: (1) a preliminary resource and needs assessment was conducted among agencies serving farmworkers, (2) a GIS product was created using data available from state agencies, and (3) the GIS product was evaluated by an advisory board of qualified occupational health and safety representatives for appropriateness, applications, and ease of use. Agencies participated by sharing their available farmworker data. The GIS product consisted of a CD-ROM with data displayed in a graphic format and downloadable spreadsheet files consolidated by county demographic, crop, housing, and migrant health clinic information. Evaluators of the GIS product found it to be an accessible, unique clearinghouse for farmworker-related data. The GIS product can become a valuable tool for agencies serving farmworkers and those researching farmworker-related issues. Agencies and health professionals require useful and comprehensive databases to track and serve farmworkers, and a multi-agency partnership using GIS technology could provide this capability. Further research is required with improved definitions and resources to apply the GIS product. PMID- 15782892 TI - Simplified overturn stability monitoring of agricultural tractors. AB - Agricultural tractors are the most common source of farm work fatalities in the U.S., with overturns the most common type of incident. For the year 2001, there were 15 tractor-related fatalities in Pennsylvania, 9 of which were due to tractor rollover. A new device using low-cost sensors and microcomputers was developed around a simplified mathematical model of an agricultural tractor to inform the operator of potential tractor instability. This device communicates the current rollover potential, along with a recent history of rollover potential, to the operator of the tractor via a simple bar-graph display. The device uses a single-chip accelerometer to sense the current rollover potential and a small microprocessor to analyze the accelerometer data, compensate for variations due to temperature, and then send this information to a visual display. The use of these low-cost "off the shelf" components enabled the fabrication of a very inexpensive sensor system. Because agricultural tractors have a long service life, it was important to make the device low cost and flexible. This could enable it to be sold as an aftermarket add-on for a variety of tractor models. The device is also capable of interfacing with newer on-board tractor systems via a CAN bus to make it more attractive to tractor manufacturers who may want to incorporate this device into new models. Work is continuing on the development of an improved display to inform the tractor operator of possible instability, including display ergonomic studies, investigation of threshold levels for alerting an operator of potential instability, and investigation into audible warning signals. PMID- 15782893 TI - The role of public health improvements in health advances: the twentieth-century United States. AB - Mortality rates in the United States fell more rapidly during the late nineteenth and early twentieth centuries than in any other period in American history. This decline coincided with an epidemiological transition and the disappearance of a mortality "penalty" associated with living in urban areas. There is little empirical evidence and much unresolved debate about what caused these improvements, however. In this article, we report the causal influence of clean water technologies--filtration and chlorination--on mortality in major cities during the early twentieth century. Plausibly exogenous variation in the timing and location of technology adoption was used to identify these effects, and the validity of this identifying assumption is examined in detail. We found that clean water was responsible for nearly half the total mortality reduction in major cities, three quarters of the infant mortality reduction, and nearly two thirds of the child mortality reduction. Rough calculations suggest that the social rate of return to these technologies was greater than 23 to 1, with a cost per person-year saved by clean water of about dollar 500 in 2003 dollars. Implications for developing countries are briefly considered. PMID- 15782894 TI - Long-range trends in adult mortality: models and projection methods. AB - In the study reported here, I had two objectives: (1) to test a new version of the logistic model for the pattern of change over time in age-specific adult mortality rates and (2) to develop a new method for projecting future trends in adult mortality. A test of the goodness of fit of the logistic model for the force of mortality indicated that its slope parameter is nearly constant over time. This finding suggests a variant of the model that is called the shifting logistic model. A new projection method, based on the shifting mortality model, is proposed and compared with the widely used Lee-Carter procedure. PMID- 15782895 TI - Fifteen years later: can residential mobility programs provide a long-term escape from neighborhood segregation, crime, and poverty? AB - We examined whether the Gautreaux residential mobility program, which moved poor black volunteer families who were living in inner-city Chicago into more-affluent and integrated neighborhoods, produced long-run improvements in the neighborhood environments of the participants. We found that although all the participants moved in the 6 to 22 years since their initial placements, they continued to reside in neighborhoods with income levels that matched those of their placement neighborhoods. Families who were placed in higher-income, mostly white neighborhoods were currently living in the most-affluent neighborhoods. Families who were placed in lower-crime and suburban locations were most likely to reside in low-crime neighborhoods years later. PMID- 15782896 TI - Understanding racial differences in the economic costs of growing up in a single parent family. AB - This article examines whether the economic consequences of growing up in a single parent family differ for black children and white children. It is important to understand whether the costs differ across racial groups because although much of the rhetoric about poor single-parent families focuses on inner-city blacks, most children who live in such families are white. If the costs of living with only one parent vary across groups, then policies that are aimed at reducing the costs that do not acknowledge this variation will not target resources efficiently. We found that the economic costs of living with a single parent are larger for black children than for white children. Most of the discrepancy can be attributed to differences in remarriage rates, marital stability, welfare participation, and female labor supply. PMID- 15782897 TI - Remarks on the analysis of causal relationships in population research. AB - The problem of determining cause and effect is one of the oldest in the social sciences, where laboratory experimentation is generally not possible. This article provides a perspective on the analysis of causal relationships in population research that draws upon recent discussions of this issue in the field of economics. Within economics, thinking about causal estimation has shifted dramatically in the past decade toward a more pessimistic reading of what is possible and a retreat in the ambitiousness of claims of causal determination. In this article, the framework that underlies this conclusion is presented, the central identification problem is discussed in detail, and examples from the field of population research are given. Some of the more important aspects of this framework are related to the problem of the variability of causal effects for different individuals; the relationships among structural forms, reduced forms, and knowledge of mechanisms; the problem of internal versus external validity and the related issue of extrapolation; and the importance of theory and outside evidence. PMID- 15782898 TI - Health consequences of forest fires in Indonesia. AB - We combined data from a population-based longitudinal survey with satellite measures of aerosol levels to assess the impact of smoke from forest fires that blanketed the Indonesian islands of Kalimantan and Sumatra in late 1997 on adult health. To account for unobserved differences between haze and nonhaze areas, we compared changes in the health of individual respondents. Between 1993 and 1997, individuals who were exposed to haze experienced greater increases in difficulty with activities of daily living than did their counterparts in nonhaze areas. The results for respiratory and general health, although more complicated to interpret, suggest that haze had a negative impact on these dimensions of health. PMID- 15782899 TI - Educational selectivity in U.S. immigration: how do immigrants compare to those left behind? AB - Current immigration research has revealed little about how immigrants compare to those who do not migrate. Although most scholars agree that migrants are not random samples of their home countries' populations, the direction and degree of educational selectivity is not fully understood. This study of 32 U.S. immigrant groups found that although nearly all immigrants are more educated than those who remain in their home countries, immigrants vary substantially in their degree of selectivity, depending upon the origin country and the timing of migration. Uncovering patterns of immigrant selectivity reveals the fallacy in attributing immigrants' characteristics to national groups as a whole and may help explain socioeconomic differences among immigrant groups in the United States. PMID- 15782900 TI - The importance of education-occupation matching in migration decisions. AB - In this article, we present and test a model that incorporates education occupation matching into the migration decision. The literature on education occupation matching shows that earnings are affected by how individuals' education matches that required by their occupation. Accordingly, individuals with more schooling than required by their occupation have an additional incentive to migrate: the increase in earnings that occurs with a more beneficial education-occupation match. Using data from Mexico, we found statistical support for the importance of education-occupation matching in migration decisions. Education-occupation matching provides a plausible explanation for the mixed findings in the literature on the relationship between educational attainment and migration. PMID- 15782902 TI - Quantification of the heat exchange of chicken eggs. AB - In the incubation process of domestic avian eggs, the development of the embryo is mainly influenced by the physical microenvironment around the egg. Only small spatiotemporal deviations in the optimal incubator air temperature are allowed to optimize hatchability and hatchling quality. The temperature of the embryo depends on 3 factors: (1) the air temperature, (2) the exchange of heat between the egg and its microenvironment and (3) the time-variable heat production of the embryo. Theoretical estimates on the heat exchange between an egg and its physical microenvironment are approximated using equations that assume an approximate spherical shape for eggs. The objective of this research was to determine the heat transfer between the eggshell and its microenvironment and then compare this value to various theoretical estimates. By using experimental data, the overall and the convective heat transfer coefficients were determined as a function of heat production, air humidity, air speed, and air temperature. Heat transfer was not affected by air humidity but solely by air temperature, embryonic heat generation, and air speed and flow around eggs. Also, heat transfer in forced-air incubators occurs mainly by convective heat loss, which is dependent on the speed of airflow. A vertical airflow is more efficient than a horizontal airflow in transferring heat from the egg. We showed that describing an egg as a sphere underestimated convective heat transfer by 33% and was, therefore, too simplistic to accurately assess actual heat transfer from real eggs. PMID- 15782901 TI - Differences in disability among older women and men in Egypt and Tunisia. AB - Research on child survival and health has indicated disparities between boys and girls in selected Middle Eastern countries. Health disparities in later life are understudied in this region. In this article, we examine differences between women and men in later-life activity limitation in Egypt and Tunisia. Difficulty executing physical tasks is more common for women than for men in both study sites, although differences are smaller after adjustment for underlying illness. Differences in the difficulty of executing physical tasks also are sensitive to environmental controls in variable ways across the study sites. The findings caution against the sole use of reported disability in comparative studies of gender and aging. PMID- 15782903 TI - Utilizing different ratios of alfalfa and layer ration for molt induction and performance in commercial laying hens. AB - Molting is a common practice used by the commercial egg industry to rejuvenate flocks for a second or third laying cycle. During this time the hens rest from production, and the reproductive organs are rejuvenated to increase production and quality during the next laying cycle. Although feed withdrawal (FW) is the most popular and effective method of molt induction, it has come under scrutiny due to food safety issues and animal welfare issues. This study involved feeding alfalfa mixed with layer ration at different ratios to hens to determine their ability to induce molt. The treatment ratios were 100% alfalfa (A100), 90% alfalfa and 10% layer ration (A90), and 70% alfalfa and 30% layer ration (A70). In addition, a fully fed (FF) nonmolted control and a FW negative control were used. Alfalfa is an insoluble, high fiber feedstuff with low metabolizable energy. Egg production for A90 and FW treatments ceased completely by d 6, whereas birds fed A100 and A70 ceased egg production by d 8. Ovary and oviduct weight of hens fed all molting diets decreased (P < 0.05) by an average of 1.5 to 2.5% (BW basis) compared with FF control during the 9-d molt induction period. As the percentage of layer ration increased, feed intake also increased and percentage of BW loss decreased during the 9-d molt induction period. Hens molted by FW lost an average of 25.8% BW, whereas A70 hens lost 18.9% BW. Nonmolted hens (FF) and A70 treatment hens had significantly lower (P < 0.05) egg production when compared with all other treatments over the 39-wk postmolt period. FF treatment hens also had lower (P < 0.05) albumen heights when compared with all other treatments. From these results, alfalfa or alfalfa mixed with layer ration appears to be viable alternatives to conventional FW methods for the successful induction of molt and retention of postmolt performance. PMID- 15782904 TI - Genetic analysis on the direct response to divergent selection for phytate phosphorus bioavailability in a randombred chicken population. AB - The current study was undertaken to evaluate the direct response to 3 generations of divergent selection for phytate P bioavailability (PBA) in the Athens-Canadian randombred chicken population. Cumulated divergent response (R(C)) was measured as the line difference in PBA at a given generation after adjusting for hatch and sex effects. Results showed a significant response at generation (G)1. The R(C) was unchanged from G1 to G2 and increased (1.62%) from G2 to G3 (P < 0.01) due to the application of best linear unbiased prediction (BLUP) selection in the line selected for high PBA at G2. The average BLUP estimated breeding values were used to estimate the genetic trend for the selected trait across generations. The results showed that the genetic trend was symmetric at G1 and G2 but asymmetric at G3. The application of mixed model methodology was effective in separating the environmental component from phenotypic change. When the data of the high (H) line or the low (L) line in the selected generations (G1 to G3) were combined with the data from the base population (G0), the heritability estimates for PBA were 0.07 +/- 0.02 and 0.09 +/- 0.02, respectively. The line selected for high PBA showed gain, and the line selected for low PBA showed a decrease in estimated breeding values across the generations. The results demonstrated that modest progress could be obtained by incorporating PBA into selection programs. However, other correlated traits of economic importance need to be evaluated before any decision to incorporate selection of PBA into breeding schemes be initiated. PMID- 15782905 TI - Effect of genetic selection on MyoD and myogenin expression in turkeys with different growth rates. AB - Skeletal muscle development requires the ordered expression of specific myogenic regulatory factors, which include MyoD, Myf5, myogenin, and MRF4. The MyoD and Mrf5 factors are required for the determination of myoblasts, whereas myogenin and MRF4 play a pivotal role in terminal differentiation. In the current study, males and females of a turkey genetic line selected only for increased 16-wk BW (F line) and an unselected randombred control (RBC2 line) from which the F line was developed were used to investigate the developmental expression of MyoD and myogenin mRNA in embryonic pectoralis major muscle and myogenic satellite cells. Pectoralis major muscle was isolated at embryonic d (ED) 14, 16, 18, 20, 22, and 24. The mRNA levels of MyoD and myogenin were measured using a real-time quantitative polymerase chain reaction method. Both MyoD and myogenin expression declined during embryonic development. The decrease in MyoD expression started at ED 16 for the F line and at ED 18 for the RBC2 line for both sexes. Myogenin expression in both lines began to decline at ED 14. The F line males had lower myogenin expression at ED 14, 16, and 18 than the RBC2 line males, which was similar for the F line females compared with the RBC2 line females except there was no significant difference at ED 18. The RBC2 line males had greater expression than the females for myogenin at ED 16 and 18 for the RBC2 line. Proliferating myogenic satellite cells in both lines and sexes expressed low levels of MyoD and myogenin. After the initiation of differentiation in both lines and sexes, there was a sharp surge in MyoD expression at 24 h followed by a decrease at 48 h and then an increase in expression through 72 or 96 h of differentiation. There were line and sex differences in myogenin expression during the differentiation process. These data are suggestive of growth- and sex related differences in the expression of myogenic regulatory factors key to muscle cell proliferation and differentiation, which will affect muscle growth rate. PMID- 15782906 TI - Bt176 corn in poultry nutrition: physiological characteristics and fate of recombinant plant DNA in chickens. AB - A genetically modified Bt176 corn hybrid, which contains an insecticidal protein against the European corn borer, and its conventional, nonmodified counterpart were evaluated in 4 separate trials to verify substantial equivalence in feeding value and animal performance. Thirty-six individually kept laying hens and 3 replicates of 94 broiler chickens each, assigned to 12 cages, were fed 2 different hen and broiler diets containing either 60% conventional or 60% Bt176 corn. The nutrient compositions of the 2 corn hybrids and the 2 corn diets revealed no major differences. Furthermore, metabolism and performance data revealed no significant differences between the birds that received the conventional, nonmodified corn, and those that received the modified corn diets. The detection of the genetic modification, by PCR, in feed obtained from insect resistant Bt corn, in tissues and products from animals fed Bt corn is described. In all evaluated chicken tissues of muscle, liver, and spleen, the corn chloroplast ivr gene fragment was amplified. It can be deduced from these findings and from other studies that the transfer of DNA fragments into the body is a normal process that takes place constantly. Nevertheless, no recombinant plant DNA fragments such as recombinant bla or cry1A(b) fragments could be found. Bt-gene specific constructs from the Bt corn were not detected in any of the poultry samples, neither in organs, meat, nor eggs. PMID- 15782907 TI - Nutritive value of high-oleic acid sunflower seed for broiler chickens. AB - Two experiments were conducted to evaluate the use of high-oleic acid sunflower seed (HOASS) in broiler diets. In the first experiment, HOASS was included in a basal diet at 80, 160, 240, and 320 g/kg at the expense of the energy-yielding ingredients, and the AMEn values of the experimental diets were determined. The linear regression equation of AMEn values on rate of inclusion was calculated. Extrapolation value for the AMEn of HOASS at 100% inclusion was 4224 +/- 77 kcal/kg. In the second experiment, diets containing up to 200 g of HOASS/kg were given to broilers (Cobb) from 0 to 42 d, and performance parameters, nutrient digestibility, and AMEn value were determined at 12 and 42 d of age. Birds fed the diets containing 100 and 200 g HOASS/kg gained less weight (P < 0.001) than those fed the diet containing no HOASS at both ages. Differences in feed-to-gain ratio were only significant for the diet with the highest concentration of HOASS. Apparent digestibility of nutrients and dietary AMEn contents of diets increased with age; thus, the mean digestibility of diets for amino acid N and for total fatty acids increased from 82.1 and 68.0% at 12 d to 86.7 and 84.7% at 42 d, respectively, and AMEn content was improved by 6.5%. Inclusion of HOASS in the diet decreased the digestibilities of fat, oleic acid, and total fatty acids. A decrease in the digestibility of aspartic acid, threonine, tyrosine, valine, isoleucine, and AMEn with increasing inclusion level was also observed at 12 d of age. PMID- 15782908 TI - Predicting effective caloric value of nonnutritive factors: III. Feed form affects broiler performance by modifying behavior patterns. AB - Two trials of an experiment were conducted to confirm the relationships among effective caloric value (ECV) of the diet, net energy for gain (NEg), BW, feed conversion ratio (FCR), and broiler behavior. Further, we sought to examine such factors with benefits of pelleting, including feed form history (pellets vs. mash) in females from 2 strains of commercial broilers. Composition of gain was measured on a sample of birds in both trials. In trial 1 birds were reared to 23 d on feed in crumble form, when the birds were provided a feed in pellet or mash form for 7 d. Pelleting the feed increased ECV and total NEg, while decreasing eating and increasing resting behavior. Significant correlations (P < 0.05) among resting, NEg, and ECV occurred. In trial 2, birds were reared to 23 d on a crumble diet and then fed diet in mash or pellet form to 36 d. At 37 d of age, half of the birds from each strain and feed form history combination were switched to the alternative feed form. Interactions of strain by grower feed form were present for BW, initial fat, and body energy content indicating that pelleted feed was required for optimum broiler performance of 1 strain. Grower feed form by finisher feed form interactions were present and demonstrated that birds switched from pellets or mash to the alternate feed form consumed more feed in less time than birds that remained on their previous feed form. Significant correlations were observed in both trials between behaviors and FCR and ECV, whereas NEg reflected these differences in trial 1 but not trial 2. Regression analysis indicated that FCR and subsequently ECV were best predicted by lean gain, whereas NEg was best predicted by fat gain. Further, regression analysis established interactive equations in which ECV was predicted (R2 > 0.99) by eating and resting behavior. The results of these trials indicate that the effects of feed form are caused by a modification of behavior patterns, that ECV is responsive to such behavior changes, and that ECV is an effective estimator of the relative caloric value of genetic, management, and husbandry influences. PMID- 15782909 TI - Direct determination of optimal amino acid intake for maintenance and growth in broilers. AB - This study has used a direct approach to calculate the optimal dietary supply of amino acids for maintenance and growth in broiler chicks. Amino acid intake and accretion in the carcass in Ross 308 chicks was measured following the feeding of different experimental diets with controlled feed intake from hatching through 42 d. Increasing crude protein levels at constant amino acid:protein ratio improved growth and reducing the crude protein levels at constant amino acid:protein ratios depressed performance. Altering the protein at constant essential amino acid concentrations had little affect on growth. Plotting the amino acid intake per metabolic BW against amino acid accretion per unit of metabolic BW yielded a straight line whose slope represents the efficiency of utilization of the amino acid for growth; the intercept represents maintenance. These values were then used to calculate the amino acid intakes required for any BW and growth rate. For comparison, inputs were calculated for the growth targets recommended in the Ross manual and compared with the dietary recommendations. The measured results were similar to the overall empirical recommendations although values differed slightly, with threonine required at higher levels then recommended. Using the growth rates obtained in this trial, optimal amino acid supply was compared with the NRC (1994) recommendations. This indicated that optimal amino acid supply was higher then NRC (1994) recommendations for most amino acids. The calculation of optimal amino acid supply as a function of BW and growth will allow effective formulation of diets for maximal efficiency. PMID- 15782910 TI - Assessment of dietary amino acid scarcity on growth and blood plasma proteome status of broiler chickens. AB - Dietary Lys needs for chicks were studied. A titration diet consisting of progressive amounts of dietary Lys from 0.95% up to 1.40% was fed to broiler chicks from 0 to 18 d of age. Optimal dietary Lys level was calculated using regression analysis. Body weight gain and feed conversion were maximized at Lys levels of 1.24% (1.10% digestible) and 1.27% (1.13% digestible) of diet, respectively. Blood samples were then collected from 2 groups: birds fed the lowest Lys level and birds fed dietary Lys nearest the determined requirement level (1.25% Lys). Plasma was analyzed for protein spectra via mass spectrometry and then classified by their functional characteristics. The number of proteins was similar between the 2 samples, but there was a tendency toward increased peptides for specific proteins in plasma from chicks fed adequate Lys levels. Furthermore, after these proteins were classified, more muscle-related proteins were found in plasma samples of birds fed Lys-adequate diets. It would appear that an individual dietary amino acid deficiency does not necessarily translate into decreasing protein synthesis proportionate to body weight, but rather significant changes may be occurring within the types of proteins undergoing anabolism. In conclusion, results herein illustrate the potential for using functional genomics in nutritionally related responses of poultry. PMID- 15782911 TI - Comparative development of the small intestine in the turkey poult and Pekin duckling. AB - Turkey poults and Pekin ducklings hatch from eggs of similar weights and have the same incubation periods and body weights at hatch. The male Pekin duckling, however, can attain a market weight of 3.2 kg in approximately 6 wk, whereas at the same age, male turkeys only weigh approximately 2.1 kg. For this study, fertile turkey eggs (n = 400, mean weight: 87.2 g, range: 85 to 89.9 g) and Pekin duck eggs (n = 565, mean weight: 88.6 g, range: 85 to 92.0 g) were weighed and incubated. Embryos and hatchlings were sampled during the last week of incubation, at hatch, and through 7 d of age. Yolk-free BW of poults were 2.7 g heavier than ducklings at hatch. Yolk-free BW of ducklings, however, were greater than poults at 1 d of age (P > or = 0.06), and by 7 d of age ducklings were 140 g heavier (P < or = 0.01). Yolk sac weight was similar at 21 and 25 d of incubation, yet was significantly lower in ducks at hatch, 1, and 2 d of age (P < or = 0.05). In the duckling, jejunum and ileum weights (3.7x heavier), length (1.6x longer), and density (g/cm; 2.3x more dense) were consistently heavier than in the turkey from hatch through 7 d (P < or = 0.01). Histological sections of the distal jejunum revealed more rapid villus growth in the duck from 0 to 3 d of age. The combination of increased intestinal growth (weight and length) and maturation (villus length) allowed ducks to achieve an additional 143 g of BW gain during the critical hatch through 7 d of growth. PMID- 15782912 TI - Pulmonary hypertensive responses of broilers to bacterial lipopolysaccharide (LPS): evaluation of LPS source and dose, and impact of pre-existing pulmonary hypertension and cellulose micro-particle selection. AB - Previous studies demonstrated that bacterial lipopolysaccharide (LPS, endotoxin) triggers pulmonary vasoconstriction leading to pulmonary hypertension (PHS, ascites) in broilers. The lungs of broilers are constantly challenged with LPS that can trigger pulmonary vasoconstriction. Among broilers from a single genetic line, some individuals respond to LPS with large increases in pulmonary arterial pressure (PAP), whereas others fail to exhibit any response to the same supramaximal dose of LPS. In the present study we evaluated the impact of a variety of factors on the magnitude of the PAP response of male broilers to LPS, including: (1) the role of the initial PAP (low vs. high initial PAP); (2) the source of the LPS (Salmonella typhimurium vs. Escherichia coli); (3) the dose of LPS (0.02, 0.1, and 0.5 mg/kg of BW); and (4) the role of micro-particle selection for improved pulmonary vascular capacity (cellulose survivors vs. saline-injected controls). Broilers in the low initial PAP group (21 +/- 0.34 mmHg, mean +/- SEM) did not differ in their pulmonary hypertensive response to LPS compared with broilers in the high initial PAP group (29 +/- 0.55 mmHg, mean +/- SEM). Lipopolysaccharide from S. typhimurium elicited pulmonary hypertensive responses qualitatively similar to those elicited by E. coli LPS. A detailed evaluation revealed that an LPS dose of 0.1 mg/kg of BW elicits a maximal pulmonary hypertensive response in male broilers, and broilers selected by micro particle injection for a robust pulmonary vascular capacity did not differ in their pulmonary hypertensive response to LPS compared with unselected broilers. This research confirms that the variable pulmonary hypertensive responses among broilers cannot be attributed to the source or dosage of LPS, or to differences in the baseline pulmonary arterial pressure or micro-particle selection before injecting LPS. These findings are consistent with the hypothesis that innate rather than acquired variability may influence the profile of chemical mediators released during the inflammatory cascade. PMID- 15782913 TI - Pulmonary and systemic hemodynamic responses to intravenous prostacyclin in broilers. AB - The eicosanoid vasodilator prostacyclin (PGI2) reduces resistance to pulmonary blood flow and attenuates pulmonary hypertension in mammals. However, sparse information is available regarding the responsiveness of the avian pulmonary vasculature to PGI2. Accordingly, in 3 experiments we evaluated the pulmonary vascular responses to PGI2 in male broilers. In experiment 1, infusing PGI2 (10 microg/min) into clinically healthy broilers did not reduce their pulmonary vascular resistance (PVR) but did reduce their pulmonary arterial pressure (PAP) by lowering their cardiac output. Within 4 min after stopping the PGI2 infusion, the cardiac output and PAP returned to preinfusion levels. In experiment 2, the responses to PGI2 were evaluated after arachidonic acid (AA) had been infused to preconstrict the pulmonary vasculature. The AA infusion (400 microg/min) consistently triggered dramatic, sustained pulmonary vasoconstriction (increased PVR) and pulmonary hypertension (increased PAP). Concurrent PGI2 infusions did not reduce PVR but did reduce PAP by lowering cardiac output. Within 4 min after stopping the PGI2 infusion, PAP and cardiac output returned to their previous (hypertensive) levels attributable to the ongoing AA infusion. In experiment 3, PGI2 was infused (10 microg/min) into clinically healthy (PAP < or = 24 mmHg) or subclinically hypertensive (PAP > or = 27 mmHg) broilers. Throughout this experiment broilers in the hypertensive group had higher PAP values than broilers in the healthy group. The PGI2 infusion reduced PAP in both groups but did not reduce PVR. Instead, the pulmonary hypotensive response to PGI2 infusion was associated with a reduction in cardiac output in both groups. In all 3 experiments PGI2 reduced PAP by reducing cardiac output rather than by reducing PVR. There was no evidence that PGI2 acts as an effective pulmonary vasodilator in broilers regardless of whether their pulmonary vasculature was apparently normal (clinically healthy), had been pharmacologically preconstricted (AA infusion), or initially exhibited the vasoconstriction that is typical of the pathogenesis of pulmonary hypertension syndrome in broilers (PAP > or = 27 mmHg). The consistent failure of PGI2 to elicit pulmonary vasodilation in this study suggests fundamental differences in AA metabolism or the etiology of pulmonary hypertension may exist when broilers are compared with mammals. PMID- 15782914 TI - Relationships among prehatch and posthatch physiological parameters in early nutrient restricted broilers hatched from eggs laid by young breeder hens. AB - The objective of this study was to establish relationships among various physiological parameters in early (through 72 h posthatch) nutrient restricted broiler chicks from young breeder hens. Despite a 19% decrease in mean BW between 0 and 72 h posthatch, there were no chick mortalities. Liver glycogen peaked at 24 h, and peaks in rectal temperature, plasma glucose, and plasma refractive index occurred at 48 h. By 24 h, negative correlations were noted between BW and relative liver weight and between liver glycogen and both refractive index and rectal temperature. By 24 h, positive correlations were noted between relative liver weight and liver glycogen, BW and plasma glucose, rectal temperature and plasma glucose, and rectal temperature and body fat loss score. Mean daily relative egg weight loss between 0 and 19 d of incubation (RIEWL) was positively correlated with rectal temperature at 0 h but was negatively correlated with rectal temperature at 24 h and liver glycogen at 24 and 48 h. Furthermore, mean relative hatching chick weight (RHCW) had significantly positive correlations with plasma glucose at 6 h and rectal temperature at 48 and 72 h. Broilers having a greater RHCW after hatching from eggs with a lower RIEWL may maintain higher metabolic rates between 24 and 72 h posthatch despite nutrient deprivation. However, broiler chicks from eggs exhibiting a higher than optimal RIEWL may have greater dependence on gluconeogenesis, thus requiring stricter management during initial brooding. PMID- 15782915 TI - Myosin heavy chain isoform expression is not altered in the pectoralis major muscle in selenium-deficient chickens recovering from exudative diathetic myopathy. AB - The recovery of broiler chickens experiencing skeletal muscle myopathy caused by a selenium deficiency was compared with control broiler chickens in an age matched study by ultrastructural analysis of the pectoralis major (PM) muscle and examination of the temporal expression of the developmental fast skeletal myosin heavy chain (MyHC) isoforms. Selenium-deficient chicks showing signs of exudative diathesis (ED) were injected subcutaneously with sodium selenite in water and allowed to recover. At 0, 2, 5, 10, 20, and 30 d after selenium injection, a sample of the PM muscle was removed from selenium-deficient and control chicks for analysis. Ultrastructural analysis revealed vacuolization in the PM of selenium-deficient chicks with little or no visible damage to the sarcomere. Relative amounts of chicken ventricular, embryonic, neonatal, and adult fast skeletal MyHC isoforms were determined using chicken fast skeletal MyHC isoform specific monoclonal antibodies. The temporal expression of the developmental MyHC isoforms was similar in all chickens (P > 0.05). There was no expression of chicken ventricular MyHC observed in the PM of either group. These results indicate that chicken fast muscle recovering from exudative diathetic myopathy does not use the same pathways as chicken skeletal fast muscle regenerating from physical or toxic injury in which temporal expression of the MyHC isoforms is initially predominantly ventricular, then predominantly embryonic, neonatal, and finally predominantly adult developmental MyHC isoform. PMID- 15782916 TI - Comparison of bruises and mortality, stress parameters, and meat quality in manually and mechanically caught broilers. AB - A field trial was conducted to compare manual catching of broilers with a mechanical catching method. Both methods were compared with respect to the incidence of bruises and dead on arrival, stress parameters, and meat quality. Also the dynamics of corticosterone, glucose, and lactate were investigated on the day broilers were killed. The broilers originated from 8 commercial broiler farms; visits were made on the day of catching during spring and autumn of 2001. Broilers of one house were caught manually, and those of the second house were caught mechanically. Plasma samples were taken before catching started, 30 min after the start of catching, 30 min before the end of catching, and at exsanguination of broilers from the first- and last-loaded transport vehicles. Postmortem measurements of pH, temperature, and water-holding capacity were made. Mechanical catching was associated with higher DOA percentages than manual catching in spring, although the difference was not significant in autumn. Catching method did not influence the percentage of bruises or meat quality. Moreover, corticosterone levels indicated that both methods induced the same amount of stress. The dynamics of corticosterone, glucose, and lactate levels showed a similar pattern. Plasma levels increased at the start of catching, and they further increased during transport, shackling, and stunning. However during catching itself, no large changes were observed. Our findings indicated that attempts to reduce stress in broilers during the last day of life could better be focused on factors other than catching. PMID- 15782917 TI - Recovery of salmonellae following pH adjusted pre-enrichment of broiler carcasses treated with trisodium phosphate. AB - Trisodium phosphate (TSP) has been reported to decrease the recovery of salmonellae from processed poultry carcasses. It has been suggested that the high pH and detergent-like properties of TSP solutions are responsible for the reduction in salmonellae recovery. This project was conducted to determine if controlling pH during salmonellae pre-enrichment alters the effect of TSP on salmonellae recovery. Carcasses were obtained from a commercial processing plant immediately after the final inside-outside carcass washer, prior to any other antimicrobial treatments, and before chilling. Carcasses were assigned to 1 of 4 treatment groups: (1) TSP and alkaline pre-enrichment, (2) TSP and neutral pre enrichment, (3) non-TSP and alkaline pre-enrichment, 4) non-TSP and neutral pre enrichment. Carcasses were placed into plastic bags with 500 mL of buffered peptone water (with or without pH adjustment) and shaken for 1 min. Preincubation pH of the rinsate was measured. Carcasses were incubated in the rinse at 37 degrees C for 24 h, and incidence of salmonellae was determined. The pH of the preincubation rinsate was 8.4 for the TSP alkaline pre-enrichment, 7.2 for the TSP neutral pre-enrichment, 8.6 for the non-TSP alkaline pre-enrichment, and 7.1 for the non-TSP neutral pre-enrichment. Salmonellae were detected from 40% of the TSP alkaline pre-enrichment carcasses, 44% of the TSP neutral pre-enrichment carcasses, 54% of the non-TSP alkaline pre-enrichment carcasses, and 38% of the non-TSP neutral pre-enrichment carcasses. Neither TSP treatment nor pre enrichment pH adjustment significantly influenced carcass salmonellae detection. PMID- 15782918 TI - Research note: Functionality of electrically stimulated broiler breast meat. AB - Postmortem electrical stimulation (ES) tenderizes meat by acceleration of adenosine triphosphate (ATP) depletion, pH decline, and physical disruption of muscle fibers. It has been demonstrated that rigor development at elevated temperatures, as with slow chilling, can cause meat to develop pale color and poor water-holding capacity. The objective of this study was to compare the functionality of broiler breast meat from control and electrically stimulated carcasses with and without normal rapid chilling. Broilers were either electrically stimulated (450 mA, 450 V, 2 s on, 2 s off for 7 pulses) immediately after bleeding or used as nonstimulated controls. The ES birds were either chilled immediately (ES2) or had chilling delayed for 2 h at room temperature (ESD2). All ES breast fillets were harvested at 2 h postmortem. The control carcasses were chilled immediately and had fillets harvested at 2 h postmortem (C2) or at 8 h postmortem (C8). Electrical stimulation accelerated pH decline and prevented toughening when breast meat was deboned at 2 h postmortem, regardless of chilling rate. The water released from the gels during cooking was higher for the ESD2 than the ES2 group, which was not different from the C2 group, suggesting that ES followed by slow chilling reduced water-holding capacity compared with the ES2 and C2 groups. There were no differences in expressible moisture, gel strength, or lightness among the ES2, ESD2, and C2 treatments. These results indicated that high voltage ES followed by normal chilling did not impair protein functionality or cause pale, soft, exudative meat. However, there was some evidence that slow chilling after ES may negatively affect some water holding properties of the meat. PMID- 15782919 TI - Egg geometry calculation using the measurements of length and breadth. AB - Egg geometrical calculations that include estimations of volume and surface areas are important for the poultry industry and in biological studies, as they can be used in research on population and ecological morphology, and to predict chick weight, egg hatchability, shell quality characteristics, and egg interior parameters. The research reported here is directed at the prediction of egg volume and surface area, and develops a formula that could be both accurate and suitable for calculations. The objective of this research was to improve the accuracy of the calculations of egg volume and surface area based on the measurements of the egg length and breadth. The experiment was carried out with 90 fresh eggs from a flock of Hy-Line Brown chicks at the age of 65 wk. The resulting formula for egg volume, V, was V = (0.6057 - 0.0018B)LB2 in which L is the egg length in millimeters, and B is the egg maximum breadth in millimeters. Egg surface area, S, was calculated as S = (3.155 - 0.0136L + 0.0115B)LB, in which both L and B are taken in millimeters. PMID- 15782921 TI - A comparison of fitting growth models with a genetic algorithm and nonlinear regression. AB - A genetic algorithm (GA), an optimization procedure based on the theory of evolution, was compared with nonlinear regression for the ability of the 2 algorithms to fit the coefficients of poultry growth models. It was hypothesized that the nonlinear approach of using GA to define the parameters of growth equations would better fit the growth equations than the use of nonlinear regression. Two sets of growth data from the literature, consisting of male broiler BW grown for 168 and 170 d, were used in the study. The growth data were fit to 2 forms of the logistic model, the Gompertz, the Gompertz-Laird, and the saturated kinetic models using the SAS nonlinear algorithm (NLIN) procedure and a GA. There were no statistical differences for the comparison of the residuals (the difference between observed and predicted BWs) of growth models fit by a GA or nonlinear regression. The plotted residuals for the nonlinear regression and GA-determined growth values confirmed observations of others that the residuals have oscillations resembling sine waves that are not represented by the growth models. It was found that GA could successfully determine the coefficients of growth equations. A disadvantage of slowness in converging to the solution was found for the GA. The advantage of GA over traditional nonlinear regression is that only ranges need be specified for the parameters of the growth equations, whereas estimates of the coefficients need to be determined, and in some programs the derivatives of the growth equations need to be identified. Depending on the goal of the research, solving multivariable complex functions with an algorithm that considers several solutions at the same time in an evolutionary mode can be considered an advantage especially where there is a chance for the solution to converge on a local optimum when a global optimum is desired. It was concluded that the fitting of the growth equations was not so much a problem with the fitting methodology as it is with the form of the equation. PMID- 15782920 TI - Effect of dietary phosphorus and phytase levels on the reproductive performance of large white turkey breeder hens. AB - An experiment was conducted to determine the effect of dietary P levels and dietary phytase enzyme (E) inclusion on Large White turkey breeder hen reproductive performance from 31 to 62 wk of age. Hens were placed in a curtain sided house with 48 pens (10 birds per pen; 8 pens per treatment) at 31 wk and were fed a breeder ration with treatments as follows: HP, dietary available P = 0.55%; HPE, HP + E; MP, dietary available P = 0.35%; MPE, MP + E; LP, dietary available P = 0.17%; and LPE, LP + E. Feed and water were available ad libitum for 28 wk of lay. Diets were fed in mash form, and all other nutrients were formulated to meet or exceed NRC requirements. All hens were photostimulated in January (31 wk) with 15.5 h of light daily. Production data were recorded on a pen basis. Individual bird BW and feed consumption, by pen, were determined at monthly intervals from 31 to 62 wk. Hens were observed for weekly reproductive performance for hen housed egg production, hen-day egg production, settable eggs, cumulative settable eggs, hens out of lay, and hen mortality and for biweekly performance for egg fertility, hatchability of all eggs, hatchability of fertile eggs, egg weight loss, conductance, conductance constant (k), and embryonic mortality. Egg weight, eggshell thickness, egg components, and albumen and yolk P were measured monthly. At 62 wk of age, hen tibia P, plasma P, total fecal P, and water-soluble fecal P were determined. Decreasing dietary P resulted in no decreases in reproductive performance for turkey breeder hens to 62 wk. Additionally, decreased dietary P resulted in decreased total fecal P and water soluble fecal P. Feeding turkey breeder hens dietary phytase enzyme resulted in significantly fewer hens going out of lay; however, this was not reflected in hen housed egg production. It was concluded that phosphorus could be lowered in turkey breeder hen diets, compared with current surveyed industry levels, without impairing reproductive performance. PMID- 15782923 TI - The psychological and social burdens of psoriasis. AB - Psoriasis imposes psychological and social burdens on sufferers, in addition to the physical toll. A recent survey of 502 people with moderate-to-severe psoriasis discovered that 38% of patients with psoriasis experience disease related disruption in day-to-day activity, including work, school, interpersonal relationships, recreational activities, and intimacy. Discrimination, lack of self-confidence, and feelings of isolation, anger, and hopelessness are also quite common. In addition, another survey of 1000 adult Americans without psoriasis was conducted to determine the awareness of psoriasis by the general public. This survey confirmed a low level of awareness and understanding of the disease among people without psoriasis, suggesting that a lack of sensitivity may contribute to the social burdens of suferers. PMID- 15782922 TI - Evaluation of transgenic hybrid corn (VIP3A) in broiler chickens. AB - A 49-d feeding study evaluated whether standard broiler diets prepared with Syngenta Seeds VIP3A transgenic derived corn grain had any unanticipated adverse effects on male or female broiler chickens as compared with diets prepared with nontransgenic (isoline) control corn grain. Two commercial lots of grain grown in North Carolina during the 1999 (NC 1999) and 2000 (NC 2000) seasons were included for reference purposes. Broiler growth was excellent with males reaching 3466 g and females reaching 2882 g at 49 d of age. Final BW of the VIP3A, isoline, and NC 1999 corn groups were within 21.1 g, whereas the NC 2000 group was 42.4 g lower than the lowest of this group. There was no overall corn source effect on adjusted feed conversion ratio (FCR) or mortality to 49 d of age. Carcass analysis demonstrated no differences in percentage yield due to corn source among males and females other than percentage wings in females. Comprehensive clinical chemical analyses of blood taken from representative birds at 49 d of age showed no differences due to corn sources. The transgenic VIP3A hybrid diets numerically supported the most rapid broiler chicken growth, the second lowest mortality rate and best FCR, without practical differences in carcass yield. The few differences found in this study were not unique to a given corn source but instead appeared to be distributed equally across the diet groups evaluated in the study. Although it was not clear whether small differences in performance were attributable to the transgenic corn per se or were due to possible slight differences in overall composition of the formulated diets, it was clear that the transgenic corn had no deleterious effects on broiler performance and carcass yield in this study. PMID- 15782924 TI - Experiences and influences of women with cosmetic tattooing. AB - Cosmetic tattooing (permanent makeup of eyebrows, eyeliner, or lipliner) procedures are flourishing among women worldwide. Recurrent themes provide knowledge and understanding so dermatology nurses can aid in their procedural decision making. PMID- 15782925 TI - What's your assessment? Psoriasis with Kapposi sarcoma. PMID- 15782926 TI - Patient education and morbidity in atopic eczema. AB - Effective treatment of atopic eczema is dependent upon good management by the patients and parents. It is possible to encourage patients and improve their compliance through nurse consultation and Web-based information. PMID- 15782927 TI - Giant basal cell carcinoma. PMID- 15782928 TI - Epidermolysis bullosa: management principles for the neonate, infant, and young child. PMID- 15782929 TI - Quality of life--part II: using the holistic caring praxis in skin and wound care. PMID- 15782930 TI - Anatomy and physiology of the skin. PMID- 15782931 TI - The primary and secondary B-cell response. PMID- 15782932 TI - On leadership: attending, questioning and quality. AB - Health care is just now feeling the effects of many years of neglect of serious attention to quality outcomes. We have many tools available now to change our thinking and to provide techniques to attain excellence in quality, such as Six Sigma and principals from Toyota. However, these techniques will only get us to a minimal level of quality. We need to change our questions, think the impossible, and break out of our old modes of thinking about quality. Zero defects are possible. We only have to take that quantum leap and think in new ways. We are posed on the edge of a quantum leap into new ways of thinking about quality. We will hear much more about nurse-sensitive indicators in the near future. Nurse leaders should change their questions now to attend to new models that will get us closer to the ultimate "impossible thinking" goal of zero defects in nurse sensitive indicators of quality. PMID- 15782934 TI - Application of ITS sequence analysis, RAPD and AFLP fingerprinting in characterising the yeast genus Fellomyces. AB - Three molecular techniques, ITS sequence analysis, random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) were used to study phylogenetic and genotypic relationships among strains of the genus Fellomyces. In the analyses were included strains isolated predominantly from epiphytic lichens collected in Indonesia, China and Mexico. The polyphasic approach indicated that the Fellomyces isolates are genotypically heterogeneous and that lichens represent a specific environment for selection of large number of the sterigmatoconidia producing species. The phylogenetic and genotypic analysis confirmed the existence of 11 currently accepted Fellomyces species and indicated that several species may be the new representatives of the genus. The RAPD and AFLP analyses demonstrated a higher potential in distinguishing the Fellomyces strains than the ITS regions. Since the sequence analysis showed low or no divergence among several strains, both RAPD and AFLP fingerprinting indicated that the strains may be discriminated at the species level. PMID- 15782933 TI - Fellomyces mexicanus sp. nov., a new member of the yeast genus Fellomyces isolated from lichen Cryptothecia rubrocincta collected in Mexico. AB - Two strains isolated from the lichen Cryptothecia rubrocincta collected in Mexico are described as new members of the sterigmatoconidia-producing genus Fellomyces. Based on the identical sequences of the D1/D2 regions of 26S rDNA, as well as the results of the cluster analysis of the AFLP fingerprints, the strains have been shown to be conspecific. The 26S rDNA-based phylogeny has indicated that Fellomyces mexicanus clusters within a clade including species such as four other Fellomyces isolates from lichens: F. chinensis, F. lichenicola, F. sichuanensis and F. thailandicus. F. mexicanus is characterised by the presence of xylose in the cell walls and CoQ10 in the mitochondrial membrane. The type culture is strain HB25=CBS8279. PMID- 15782935 TI - Environmental pH as an important factor for the distribution of urease positive ammonia-oxidizing bacteria. AB - The effect of pH on ureolytic activity of a number of chemolithotrophic ammonia oxidizing bacteria (AOB) has been studied in context with distribution patterns of these species. The pH-optima for urea-based nitrification were found to differ clearly among the examined species. Pronounced optima ranged between pH 5.0 and 8.0. Urease is an intracytoplasmic enzyme and should therefore be independent of the external pH. Our first results indicated the presence of a pH-dependent uptake system for urea. Simultaneous oxidation of free ammonia, possible only at high pH values, led to a strong intensification of ureolysis. The lag-phase of growth on urea as the sole energy source was clearly prolonged compared to free ammonia. Our results point on the existence of an active, most likely energy linked urea-uptake system in addition to a possible passive diffusion of urea. The different pH-optima of urea-uptake agree with known distribution patterns of distinct AOB. It might be a reason for the shift of dominant Nitrosospira populations along pH gradients in acid soils as observed by others in molecular analyses of natural AOB populations. PMID- 15782936 TI - Molecular typing of Klebsiella pneumoniae isolates from public hospitals in Recife, Brazil. AB - Thirty nosocomial isolates of Klebsiella pneumoniae, collected from three public hospitals in Recife, Brazil, between 1999 and 2000, were analysed in order to determine their epidemiological relatedness and genetic characteristics. The isolates were characterised by biotyping, antibiotyping, protein analysis, plasmid profile and random amplified polymorphic DNA (RAPD). The majority of the clinical isolates were resistant to multiple antibiotics, in particular to beta lactams, and 30% were found to be ESBLs producers. RAPD proved to be the most effective technique in discriminating unrelated K. pneumoniae isolates. It was confirmed by the highly genetic similarity found among related isolates from an hospital outbreak. We conclude that K. pneumoniae infections in Recife has been caused by a variety of bacterial genotypes. This is the first report that revealed the subtypes of K. pneumoniae in Brazil by plasmid analysis and RAPD. PMID- 15782937 TI - Diversity of ectomycorrhizal fungi naturally established on containerised Pinus seedlings in nursery conditions. AB - The study examined the diversity of ectomycorrhizal fungi, naturally established on roots of containerised Pinus seedlings in a nursery, using PCR-RFLP and sequencing of the nuclear ribosomal internal transcribed spacer. Seventy-two samples, including ectomycorrhizae and fruit bodies, were examined. Molecular typing assigned the fungal symbionts to four ectomycorrhizal Boletales: Rhizopogon rubescens, Suillus bovinus, S. variegatus, and R. luteolus. R. rubescens was abundant (37.5%), while Suillus and R. luteolus species were moderately established (25-26%) and rare (2.8%), respectively. In addition, Rhizopogon species colonised P. nigra ssp. salzmannii seedlings, whereas Suillus species were identified on Pinus nigra ssp. nigra seedlings. The diversity and the ability of these naturally established symbionts under artificial nursery conditions were discussed. The molecular survey investigated here should contribute to successful monitoring of mycorrhizal application under both nursery and plantation conditions. PMID- 15782939 TI - Polymerase chain reaction for identification of herpes simplex virus (HSV-1), cytomegalovirus (CMV) and human herpes virus-type 6 (HHV-6) in oral swabs. AB - Considering that sensitive and specific methods to detect HSV-1, CMV and HHV-6 on oral mucosa have a great impact on oral diagnosis practice and research, together with the evidence that PCR is a rapid and reliable method, the purpose of the present study was to develop primer sets to detect HSV-1, CMV and HHV-6 in oral swabs by nested polymerase chain reaction (nested PCR). We developed a practical method for sample collection without tissue trauma, and the swabs were stored until used for DNA extraction. After the nested PCR a DNA fragment of 241 bp corresponding to HSV-1 was amplified. DNA fragments of 224 and 369 bp were amplified corresponding to CMV and HHV-6, respectively. DNA sequencing analysis confirmed the expected sequences of each virus. In conclusion, it was demonstrated that these new primer sets are able to identify HSV-1, CMV and HHV-6 in oral swab using nested PCR. PMID- 15782938 TI - Arg169 is essential for catalytic activity of 3-hydroxybenzoate 6-hydroxylase from Klebsiella pneumoniae M5a1. AB - 3-Hydroxybenzoate 6-hydroxylase from Klebsiella pneumoniae M5a1 is an enzyme that utilizes 3-hydroxybenzoate (3-HBA) as substrate yielding gentisate. Site-directed mutagenesis was carried out to define which residues may be involved in catalytic reaction. Substitution of arginine to glutamate at position 169 of the enzyme resulted in the complete loss of catalytic activity. This indicated Arg169 may play an important role in 3-HBA 6-hydroxylase catalysis. PMID- 15782940 TI - Isolation, purification, and identification of the virulence protein VirE2 from Agrobacterium tumefaciens. AB - Bacteria of the genus Agrobacterium can transfer a portion of their Ti plasmid (T DNA) in complex with the VirE2 and VirD2 proteins into the plant-cell nucleus and cause it to be integrated in the host-cell chromosomes. The mechanism of T-DNA transfer across the plant-cell membrane and cytoplasm is unknown. The aim of this study was to isolate the virulence protein VirE2 in order to explore its role in T-DNA transfer across the eukaryotic-cell membrane and cytoplasm. To obtain VirE2, we cloned the virE2 gene into plasmid pQE31 in Escherichia coli cells. VirE2 protein was isolated from E. coli XL-1 blue cells containing a recombinant plasmid, pQE31-virE2. The cells were ultrasonically disrupted, and the protein containing six histidine residues at the N-terminal end was isolated by affinity chromatography on Ni-NTA agarose. The purified preparation was tested by immunodot, by using polyclonal rabbit antibodies and miniantibodies produced toward VirE2. The capacity of the recombinant protein VirE2 for interacting with single-stranded DNA was tested by the formation of complexes, recorded by agarose gel electrophoresis. In summary, A. tumefaciens virulence protein VirE2, capable of forming a complex with single-stranded T-DNA during transfer into the plant cell, was isolated, purified, and partially characterized. Anti-VirE2 miniantibodies were obtained, and direct labeling of VirE2 with colloidal gold was done for the first time. PMID- 15782942 TI - Role of extracytoplasmic leucine rich repeat proteins in plant defence mechanisms. AB - Plant-pathogen interactions involve highly complex series of reactions in disease development. Plants are endowed with both, resistance and defence genes. The activation of defence genes after contact with avirulence gene products of pathogens depends on signals transduced by leucine-rich repeats (LRRs) contained in resistance genes. Additionally, LRRs play roles for various actions following ligand recognition. Polygalacturonase inhibiting proteins (PGIPs), the only plant LRR protein with known ligands, are pectinase inhibitors, bound by ionic interactions to the extracellular matrix (ECM) of plant cells. They have a high affinity for fungal endopolygalacturonases (endoPGs). PGIP genes are organised in families encoding proteins with similar physical characteristics but different specificities. They are induced by infection and stress related signals. The molecular basis of PG-PGIP interaction serves as a model to understand the evolution of plant LRR proteins in recognising non-self-molecules. Extensins form a different class of structural proteins with repetitive sequences. They are also regulated by wounding and pathogen infection. Linkage of extensins with LRR motifs is highly significant in defending host tissues against pathogen invasion. Overexpression of PGIPs or expression of several PGIPs in a plant tissue, and perhaps manipulation of extensin expression could be possible strategies for disease management. PMID- 15782941 TI - Diffusible and volatile compounds produced by an antagonistic Bacillus subtilis strain cause structural deformations in pathogenic fungi in vitro. AB - An efficient antagonistic strain of Bacillus subtilis, originally isolated from the rhizosphere of established tea bushes, was found to cause structural deformities in six pathogenic fungi under in vitro culture conditions. This effect was attributed to the production of diffusible and volatile antifungal compounds. Out of the selected test fungi four were phytopathogenic, while the remaining two were of clinical importance. The bacterial strain successfully restricted the growth of all test fungi in dual cultures, and induced morphological abnormalities such as mycelial and conidial deviations. The inhibitory effect caused by volatiles was greater than that by diffusible compounds. PMID- 15782944 TI - Chromatographic media for bioseparation. AB - Bioseparation processes are dominated by chromatographic steps. Even primary recovery is sometimes accomplished by chromatographic separation, using a fluidized bed instead of a fixed bed. In this review, the action principles, features of chromatography media regarding physical and chemical properties will be described. An attempt will be made to establish categories of different media. Characteristics for bioseparation are the large pores and particle sizes. To achieve sufficient capacity for ultralarge molecules, such as plasmids or nanoparticles, such as viruses monoliths are the media of choice. In these media, the mass transport is accomplished by convection, and thus, the low diffusivity can be overcome. Common to all modern chromatography media is the fast operation. There are examples where a residence time of less then 3 min, is sufficient to reach the full potential of the adsorbent. PMID- 15782943 TI - Antibacterial property of the antipsychotic agent prochlorperazine, and its synergism with methdilazine. AB - The antipsychotic drug prochlorperazine was screened in vitro for possible antimicrobial property against 157 strains of bacteria, belonging to gram positive and gram negative genera. The minimum inhibitory concentration (MIC) of prochlorperazine was determined by agar dilution method, which ranged from 25 to 200 microg/ml with respect to most of the strains. Based on such findings, a further study was undertaken to determine whether the efficacy of this drug could be enhanced in the presence of an antihistaminic agent methdilazine, reported to have remarkable antimicrobial action. Four bacterial strains, sensitive to prochlorperazine as well as to three antibacterial chemotherapeutics, viz., methdilazine, fluphenazine and thioridazine were chosen. Disc diffusion tests with prochlorperazine and methdilazine revealed marked synergism between the combination, compared to their individual effects. The synergism was found to be statistically significant (p<0.01). To assess the degree of synergism, the checkerboard analysis was performed. The FIC index of this combination turned out to be 0.37, which confirmed synergism. Therefore, this synergistic drug combination might open a new therapeutic approach to combat drug-resistance in bacterial infections. PMID- 15782945 TI - Short monolithic beds: history and introduction to the field. AB - The history of the development of short monolithic beds is described. PMID- 15782946 TI - Affinity processes realized on high-flow-through methacrylate-based macroporous monoliths. AB - The technology for preparation of rigid macroporous polymers suggested in the late 1980s has become a powerful instrument for the development of a novel scientific and practical field. At present, monolithic stationary phases are widely used in the processes of bioseparation (chromatography), bioconversion (enzyme reactors) as well as in other processes based on interphase mass distribution (for example, solid phase peptide and oligonucleotide synthesis). Bioaffinity modes of suggested dynamic methods are very promising for their use in different analytical processes (immunological, ecological, medical and other types of analytical monitoring), preparative isolation of blood proteins such as myoglobin, hemoglobin, immunoglobulins, etc. and also recombinant products directly from cell supernatants or lysates. For the first time, it has been shown that bioaffinity pairing with participation of immobilized on carefully designed rigid supports is very fast and the whole process of affinity separation can be realized within second time scale. The principle of bioaffinity recognition is generaly at the construction of biological reactors (for example, enzyme reactors). Improved kinetics of biocatalized reactions is explained by a minimal influence on the surface of the used sorbent. Very perspective field is the use of discussed monoliths for solid phase chemical synthesis of fragments of biological macromolecules (peptides and oligonucleotides). Several examples of these applications will be presented and discussed. PMID- 15782947 TI - Simple method for determining the amount of ion-exchange groups on chromatographic supports. AB - The objective of this study was to develop a fast, simple, non-destructive, non toxic and low-priced method for determining the amount of ionic groups on resins, since the conventional titration method fails to give proper results on methacrylate monoliths. After the column had been pre-saturated with a high concentration buffer solution, a low concentration buffer solution of the same pH value was pumped through the column. Measuring pH and absorbance, the profiles with a shape of typical break-through curve were obtained. It was shown that the time of the pH transient, which appeared under such conditions, could be used as a measure of the total ionic capacity of ion-exchange monolithic columns. The effect of the column length, linear velocity and varying concentrations of buffer solutions on the time of the pH transient was examined. The method was shown to be suitable for determining the amount of ionic groups on both anion and cation monolithic columns. In addition, it could also be applied to particle bed columns. The time of the pH transient and the protein dynamic binding capacity were also compared and it was concluded that for a given monolith the protein capacity can be derived from the data obtained by the new method. PMID- 15782948 TI - Affinity purification and enzymatic cleavage of inter-alpha inhibitor proteins using antibody and elastase immobilized on CIM monolithic disks. AB - Epoxy-activated monolithic CIM disks seem to be excellent supports for immobilization of protein ligands. The potential use of enzymes, immobilized on monolithic disks for rapid preparative cleavage proteins in solution was investigated. Digestion of complex plasma proteins was demonstrated by using inter-alpha inhibitors with elastase, immobilized on epoxy-activated CIM disks. Recently, a monoclonal antibody against human inter-alpha inhibitor proteins (MAb 69.31) was developed. MAb 69.31 blocks the inhibitory activity of inter-alpha inhibitor proteins to serine proteases. These results suggest that the epitope defined by this antibody is located within or proximal to the active site of the inhibitor molecule. This antibody, immobilized on monolithic disk, was used for very rapid isolation of inter-alpha proteins. The isolated complex protein was used for enzymatic digestion and isolation of cleavage products, especially from inter-alpha inhibitor light chain to elucidate precisely the target sequence for MAb 69.31 by N-terminal amino acid sequencing. Bovine pancreatic elastase immobilized on monolithic disk cleaves inter-alpha inhibitor protein complex into small fragments which are still reactive with MAb 69.31. One of these proteolytic fragments was isolated and partially sequenced. It could be shown that this sequence is located at the beginning of two proteinase inhibitor domains of the inter-alpha inhibitor light chain (bikunin). Elastase immobilized on monolithic disk offers a simple and rapid method for preparative isolation of protease cleavage fragments. The immobilized enzyme is stable and still active after repeated runs. A partial or complete digestion can be achieved by varying the flow rate. PMID- 15782949 TI - Theoretical background of short chromatographic layers. Optimization of gradient elution in short columns. AB - Although linear salt gradient elution ion-exchange chromatography (IEC) of proteins is commonly carried out with relatively short columns, it is still not clear how the column length affects the separation performance and the economics of the process. The separation performance can be adjusted by changing a combination of the column length, the gradient slope and the flow velocity. The same resolution can be obtained with a given column length with different combinations of the gradient slope and the flow velocity. This results in different separation time and elution volume at the same resolution. Based on our previous model, a method for determining the separation time and the elution volume relationship for the same resolution (iso-resolution curve) was developed. The effect of the column length and the mass transfer rate on the iso-resolution curve was examined. A long column and/or high mass transfer rate results in lesser elution volume. The resolution data with porous bead packed columns and monolithic columns were in good agreement with the calculated iso-resolution curves. Although the elution volume can be reduced with increasing column length, the pressure drop limits govern the optimum conditions. PMID- 15782950 TI - Effect of the glass surface modification on the strength of methacrylate monolith attachment. AB - The influence of glass surface modification in order to determine strength of the monolith attachment was studied. Modification consists of pre-treatment of the glass with chemicals or boiling in deionized water, silanization and drying has been investigated on different types of glass. Amount of silane groups was determined by measurement of the contact angle between the glass surface and water drop. The highest values were found for soda-lime glass. Strength of the monolith attachment was established by pumping ethanol through the monolithic capillaries and measuring the pressure drop at which monolith was dislodged. Surprisingly, it was found that the critical part of the glass surface modification procedure is glass pre-treatment. Good results were obtained with glass boiled in water for 2.5 h or more. PMID- 15782951 TI - Pressure drop in CIM disk monolithic columns. AB - Pressure drop analysis in commercial CIM disk monolithic columns is presented. Experimental measurements of pressure drop are compared to hydrodynamic models usually employed for prediction of pressure drop in packed beds, e.g. free surface model and capillary model applying hydraulic radius concept. However, the comparison between pressure drop in monolith and adequate packed bed give unexpected results. Pressure drop in a CIM disk monolithic column is approximately 50% lower than in an adequate packed bed of spheres having the same hydraulic radius as CIM disk monolith; meaning they both have the same porosity and the same specific surface area. This phenomenon seems to be a consequence of the monolithic porous structure which is quite different in terms of the pore size distribution and parallel pore nonuniformity compared to the one in conventional packed beds. The number of self-similar levels for the CIM monoliths was estimated to be between 1.03 and 2.75. PMID- 15782952 TI - Preparation and characterisation of poly(high internal phase emulsion) methacrylate monoliths and their application as separation media. AB - Poly(glycidyl methacrylate-co-ethyleneglycol dimethacrylate) monolithic supports were prepared by radical polymerisation of the continuous phase of water in oil high internal phase emulsions. Morphology of monolithic materials was studied by scanning electron microscopy and mercury intrusion porosimetry. The ratio of phase volume and the degree of crosslinking influenced the void size and pore size distribution of resulting polymers. Void sizes between 1 and 10 microm were observed and average pore sizes around 100 nm. Polymers with 60, 75, 80 and 90% pore volume were prepared and even samples with highest pore volume showed good mechanical stability. They were modified to bear weak-anion exchange groups and tested on the separation of standard protein mixture containing myoglobin, conalbumine and trypsin inhibitor. Good separation was obtained in a very short time similar to the separation obtained by commercial methacrylate monoliths. However, higher dispersion was observed. Bovine serum albumin dynamic binding capacity for monolith with 90% porosity was close to 9 mg/ml. PMID- 15782954 TI - Immobilization of deoxyribonuclease via epoxy groups of methacrylate monoliths. Use of deoxyribonuclease bioreactor in reverse transcription-polymerase chain reaction. AB - A deoxyribonuclease bioreactor was prepared by immobilization of deoxyribonuclease I through epoxy groups inherently present on poly (glycidyl methacrylate-co-ethylene dimethacrylate) monoliths. Columns with various levels of DNase activity were prepared varying immobilization temperature, pH, time and method. The apparent Michaelis-Menten constant, Km(app), and turnover number, k3app, for immobilized DNase determined by on-line frontal analysis method were, respectively, 0.28 g of DNA l(-1) and 16 dA260nm min(-1) mg(-1) of immobilized DNase. The highest activity of immobilized DNase was detected at 1 mM calcium ions concentration and mirrored properties of free enzyme; however, reaction temperature in the range from 25 to 37 degrees C has no significant effect on activity of immobilized DNase in contrary to free enzyme. The CIM DNase bioreactor was used for elimination of DNA contaminants in RNA samples prior to reverse transcription followed by PCR. PMID- 15782953 TI - Chromatographic investigation of macromolecular affinity interactions. AB - High-performance monolithic disk affinity chromatography was applied to the investigation of formation of complexes between (1) complementary polyriboadenylic and polyribouridylic acids, e.g. poly(A) and poly(U), respectively, (2) poly(A) and synthetic polycation poly(allylamine), pAA. Polyriboadenylic acid and poly(allylamine) were immobilized on macroporous disks (CIM disks). Quantitative parameters of affinity interactions between macromolecules were established using frontal analysis at different flow rates. PMID- 15782955 TI - Application of monoliths for plasmid DNA purification development and transfer to production. AB - The demand of high-purity plasmid DNA (pDNA) for gene-therapy and genetic vaccination is still increasing. For the large scale production of pharmaceutical grade plasmids generic and economic purification processes are needed. Most of the current processes for pDNA production use at least one chromatography step, which always constitutes as the key-step in the purification sequence. Monolithic chromatographic supports are an alternative to conventional supports due to their excellent mass transfer properties and their high binding capacity for pDNA. Anion-exchange chromatography is the most popular chromatography method for plasmid separation, since polynucleotides are negatively charged independent of the buffer conditions. For the implementation of a monolith-based anion exchange step into a pDNA purification process detailed screening experiments were performed. These studies included supports, ligand-types and ligand-densities and optimization of resolution and productivity. For this purpose model plasmids with a size of 4.3 and 6.9 kilo base pairs (kbp) were used. It could be shown, that up scaling to the production scale using 800 ml CIM Convective Interaction Media radial flow monoliths is possible under low pressure conditions. CIM DEAE was successfully implemented as intermediate step of the cGMP pDNA manufacturing process. Starting from 2001 fermentation aliquots pilot scale purification runs were performed in order to prove scale-up and to predict further up-scaling to 8 1 tube monolithic columns. The analytical results obtained from these runs confirmed suitability for pharmaceutical applications. PMID- 15782956 TI - Detection of processed genetically modified food using CIM monolithic columns for DNA isolation. AB - The availability of sufficient quantities of DNA of adequate quality is crucial in polymerase chain reaction (PCR)-based methods for genetically modified food detection. In this work, the suitability of anion-exchange CIM (Convective Interaction Media; BIA Separations, Ljubljana, Slovenia) monolithic columns for isolation of DNA from food was studied. Maize and its derivates corn meal and thermally pretreated corn meal were chosen as model food. Two commercially available CIM disk columns were tested: DEAE (diethylaminoethyl) and QA (quaternary amine). Preliminary separations were performed with standard solution of salmon DNA at different pH values and different NaCl concentrations in mobile phase. DEAE groups and pH 8 were chosen for further isolations of DNA from a complex matrix-food extract. The quality and quantity of isolated DNA were tested on agarose gel electrophoresis, with UV-scanning spectrophotometry, and by amplification with real-time PCR. DNA isolated in this way was of suitable quality for further PCR analyses. The described method is also applicable for DNA isolation from processed foods with decreased DNA content. Furthermore, it is more effective and less time-consuming in comparison with the existing proposed methods for isolation of DNA from plant-derived foods. PMID- 15782957 TI - Purification of genomic DNA by short monolithic columns. AB - The isolation and purification of nucleic acids is essential for many procedures in molecular biology. After showing that bacterial and eukaryotic genomic DNA can be specifically bound to the CIM DEAE monolithic column, this characteristic was exploited in development of a simple and fast chromatographic procedure for isolation and purification of genomic DNA from cell lysates that does not include the usage of toxic organic solutions. The purity and the quality of the isolate as well as the duration of the procedure was similar to other chromatographic methods used today for isolation of genomic DNA, but the initial sample volume was not restricted. PMID- 15782958 TI - Separation of pectin methylesterase isoenzymes from tomato fruits using short monolithic columns. AB - One of the main forms of tomato pectin methylesterase (PME; EC 3.1.1.1.1) that is applicable to the food industry was isolated from fresh tomato fruit. The extraction of the PME isoenzymes involved washing the fresh tomato flesh with water in order to remove sugars and than solubilizing the enzymes with a diluted HCl solution at pH 1.6. The extract was then neutralized to pH 7.4 using buffer solution. After filtration, the solution was directly fractioned using Convective Interaction Media (CIM) short monolithic disk column bearing sulfonyl (SO3) groups and using a linear gradient from 0 to 700 mM NaCl. The injection volume was 3 ml and the diameter of the column was 12 mm and length 3 mm. The isolated fractions were monitored for protein content and PME activity. The fraction with the targeted enzyme, which showed NaCl independent activity, was further purified and concentrated by ultrafiltration and finally purified by a second semi preparative cation-exchange chromatography step using a CIM carboxymethyl (CM) disk monolithic column consisting of two disks and applying a step gradient. From 1 kg of fresh tomato fruits, 7.5 mg of purified PME with molecular mass estimated to be 26 000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) was obtained. A fraction with mixed PME and polygalacturonase activity was also obtained. Compared to the published procedures for the isolation and purification of PME from plant materials, this new procedure is much faster and more efficient. The potential application of CIM disk short monolithic columns in the analysis and semi-preparative extraction and isolation of the PME isoenzyme is presented. PMID- 15782959 TI - Application of two types of CIM tube column for purification of microbial enzymes. AB - Chromatography conditions for two types of convection interaction media (CIM) tube monolithic column, DEAE-8 and C4-8, were investigated using three enzymes from different microorganisms. The enzymes were adsorbed on a CIM DEAE-8 tube column under the same conditions as conventional DEAE columns. The CIM C4-8 tube column required a high concentration of ammonium sulfate compared to the conventional C4 column for adsorbing the enzymes. The separation of enzymes on the CIM tube column chromatography was not affected at flow rates between 0.15 and 1.25 volumes of the column per min. Both columns were successfully applied to the purification of enzymes from crude enzyme solution. Thus, both CIM tube monolithic columns proved useful in greatly reducing the purification time, and could be used at any stage of enzyme purification. PMID- 15782960 TI - Choosing the right chromatographic support in making a new acetylcholinesterase micro-immobilised enzyme reactor for drug discovery. AB - The aim of the present study was to optimize the preparation of an immobilized acetylcholinesterase (AChE)-based micro-immobilized enzyme reactor (IMER) for inhibition studies. For this purpose two polymeric monolithic disks (CIM, 3 mm x 12 mm i.d.) with different reactive groups (epoxy and ethylendiamino) and a packed silica column (3 mm x 5 mm i.d.; Glutaraldehyde-P, 40 microm) were selected as solid chromatographic supports. All these reactors were characterized in terms of rate of immobilization, stability, conditioning time for HPLC analyses, optimum mobile phase and peak shape, aspecific interactions and costs. Advantages and disadvantages were defined for each system. Immobilization through Schiff base linkage gave more stable reactors without any significant change in the enzyme behaviour; monolithic matrices showed very short conditioning time and fast recovery of the enzymatic activity that could represent very important features in high throughput analysis and satisfactory reproducibility of immobilization yield. Unpacked silica material allowed off-line low costs studies for the optimization of the immobilization step. PMID- 15782961 TI - Golden rules and pitfalls in selecting optimum conditions for high-speed counter current chromatography. AB - This paper aims to be an aid to those chemists who are interested in utilizing high-speed counter-current chromatography (HSCCC), which is free of irreversible adsorption and offers high resolution comparable to column chromatography. It explains the selection of HSCCC conditions step by step including the selection of two-phase solvent systems, determination of partition coefficient (K) of analytes, preparation of two-phase solvent system and sample solution, selection of elution mode, flow rate, rotation speed, and on-line monitoring of the eluate. The paper covers both standard HSCCC and pH-zone-refining CCC techniques. Technical terms (italic) unfamiliar to the beginner are comprehensively explained in Glossary. Various examples of two-phase solvent systems used in HSCCC are listed in Appendices A and B. The commercial sources of HSCCC and other CCC instruments are described in detail in the study edited by Berthod [A. Berthod (Ed.), Counter-current Chromatography, Elsevier, Amsterdam, 2003]. PMID- 15782962 TI - High throughput processing of particulate-containing samples using supermacroporous elastic monoliths in microtiter (multiwell) plate format. AB - Two steps in parallel processing of multiple biosamples, namely, sample clarification and capture of the target protein, were integrated and combined with the direct assay of captured protein using a newly developed microtiter (96 well) plate system based on the monoliths of hydrophilic elastic supermacroporous material, cryogel. Cryogel monoliths have pore size large enough for microbial and mammalian cells to pass through unretained. Moreover, cryogel monoliths are elastic allowing them to be slightly compressed and easily introduced into the wells. When expanded, cryogel monoliths fill the well tightly with no risk of leakage in between the monolith and the walls of the well. The capillary forces keep the liquid inside the pores of the cryogel monolith making the monolith columns drainage protected. The application of a certain volume of liquid on top of a cryogel monolith column results in the displacement of exactly the same volume of liquid from the column. The concept of using supermacroporous gels in 96-well plate format offers new possibilities to the biotechnologist allowing separation of particulate matter, capturing of soluble material from particle containing media, and parallel assay of large number of non-clarified samples. PMID- 15782963 TI - Determination of flavone C-glucosides in antioxidant of bamboo leaves (AOB) fortified foods by reversed-phase high-performance liquid chromatography with ultraviolet diode array detection. AB - Reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet diode array detection (UV-DAD) was used for the simultaneous determination of four flavone C-glucosides, i.e. orientin, homoorientin, vitexin and isovitexin in several food systems fortified by the antioxidant of bamboo leaves (AOB), such as high temperature sterilized milk, sunflower seed oil and extruded rice cake for the first time. The method included extraction of flavone C-glucosides from AOB fortified foods by methanol aqueous solution, deproteinating with saturated lead acetate solution and potassium oxalate, defatting with n-hexane and clean-up by solid-phase extraction (SPE) with Phenomenex C18 cartridges. Analytes were separated with Luna C18 5 microm 250mm x 4.6mm column using acetonitrile and 1% (v/v) acetic acid (pH 3.0) as mobile phase. Good results were obtained with respect to repeatability (relative standard deviation (RSD)< 2.2%) and recovery (81.4-91.8%) which fulfilled the requirements defined by European Union (EU) legislation. The total amounts of four flavone C-glucosides were 12.56 microg/100 mL, 881.08 microg/100 mL and 1420.83 microg/100 g dry weight in AOB-fortified sterilized milk, sunflower seed oil and extruded rice cake, respectively. The method was successfully applied to the analysis of flavone C-glucosides in AOB fortified samples. The optimized procedure could also be referenced for the separation of flavone C-glucosides in other fortified foodstuffs. PMID- 15782964 TI - Rapid analysis of trace levels of antibiotic polyether ionophores in surface water by solid-phase extraction and liquid chromatography with ion trap tandem mass spectrometric detection. AB - The occurrence of antibiotics in surface and ground water is an emerging area of interest due to the potential impacts of these compounds on the environment. This paper details a rapid, sensitive and reliable analytical method for the determination of monensin A and B, salinomycin and narasin A in surface water using solid-phase extraction (SPE) and liquid chromatography-ion trap tandem mass spectrometry (LC-MS-MS) with selected reaction monitoring (SRM). Several product ions as sodiated sodium salts for MS-MS detection have been identified and documented with their proposed fragmentation pathways. Statistical analysis for determination of the method detection limit (MDL), accuracy and precision of the method is described. The average recovery of ionophore antibiotics in pristine and wastewater-influenced water was 96.0+/-8.3% and 93.8+/-9.1%, respectively. No matrix effect was seen with the surface water. MDL was between 0.03 and 0.05 microg/L for these antibiotic compounds in the surface water. The accuracy and day-to-day variation of method fell within acceptable ranges. The method is applied to evaluate to the occurrence of these compounds in a small watershed in Northern Colorado. The method verified the presence of trace levels of these antibiotics in urban and agricultural land use dominated sections of the river. PMID- 15782965 TI - Liquid chromatographic method for the micro-quantitative determination of clodinafop in soil, wheat and Phalaris minor. AB - A liquid chromatographic method was developed for the determination of clodinafop propargyl herbicide. Clodinafop-propargyl was converted to clodinafop acid by alkaline hydrolysis as clodinafop-propargyl rapidly forms bioactive clodinafop acid in soil and plant environment. Recovery methods for both the acid and ester from different matrices were standardized. The sensitivity of the method for ester and acid was 5 and 2 ng, respectively, with limits of detection of 0.5 and 0.1 microg ml(-1). The method was standardized for the determination of clodinafop residues in soil and plant samples using HPLC. The recovery of clodinafop from soil and plant samples with ethyl acetate was significantly higher (78-83%) than those with dichloromethane, toluene and methanol (60-70%). The limit of determination of clodinafop in soil and plant samples ranged between 1 and 1.2 ng g(-1). In field soil, residues of clodinafop dissipated with a half life of 3.44 days. PMID- 15782966 TI - Separation and quantification of beer carbohydrates by high-performance liquid chromatography with evaporative light scattering detection. AB - An HPLC method with an evaporative light scattering detector was optimized and validated for quantification of carbohydrates in beer. The chromatographic separation was achieved using a Spherisorb NH2, 5 microm chromatographic column and gradient elution with acetonitrile/water. The determinations were performed in the linear range of 0.05-5.0 g/L for fructose, 0.05-5.0 g/L for glucose, 0.05 15.0 g/L for maltose, 0.05-10.0 g/L for maltotriose, and 0.05-5.0 g/L for maltotetraose. The detection limits were 0.005 g/L for fructose, 0.008 g/L for glucose, and 0.01 g/L for maltose, maltotriose, and maltotetraose. The reliability of the method in terms of precision and accuracy was evaluated in three beer matrices, low alcohol beer, 6% alcohol beer, and beer made with part of adjuncts (4.5% alcohol). Relative standard deviations (RSDs) ranged between 1.59 and 5.95% (n = 10), and recoveries ranged between 94 and 98.4%. PMID- 15782967 TI - Simultaneous determination of NSO-heterocycles, homocycles and their metabolites in groundwater of tar oil contaminated sites using LC with diode array UV and fluorescence detection. AB - For monitoring groundwater at tar oil contaminated sites a simple method of analysis was developed for the simultaneous detection of several NSO heterocyclic compounds, homocyclic compounds, mobile two- and three-cyclic PAHs and selected metabolites. The groundwater samples are enriched using SPE with polymer material at pH 4. Chromatographic separation and detection is performed by LC with diode array UV or fluorescence detection. The recoveries of 25 selected compounds were mostly between 80-110% and the detection limits were 0.4-2.4 microg/L for UV detection and for the fluorescence detectable compounds 0.4-140 ng/L. The method was successfully applied to groundwater samples from a wood preserving facility. Especially benzo(b)thiophene showed an increasing dominance downgradient of the source. Detection of metabolites, such as 1-hydroxyiso-, 2-hydroxyquinoline and 2 hydroxy-4-methylquinoline, 2-naphthoic acid, and 1-indanone, indicating in situ biodegradation, was confirmed by LC-ESI-MS analysis. PMID- 15782968 TI - Molar mass distributions by gradient liquid chromatography: predicting and tailoring selectivity. AB - Interactive liquid chromatography (iLC) for polymer analysis is usually applied to the characterisation of distributions other than molar mass. In particular, its use for the determination of chemical-composition, functionality-type and tacticity distributions has been demonstrated. The application of iLC for the determination of molar mass distributions (MMDs), however, has not yet been fully explored. An expanded version of the reversed-phase liquid chromatography model has been developed to describe and predict how the retention behaviour of polydisperse polystyrene samples changes with molar mass. The relationship between molar mass and the parameters of the model has been investigated in some detail and non-linear correlations were found. From the model and the relationships between the model parameters and molar mass, calibration curves (retention time versus molar mass) were constructed to predict changes in chromatographic selectivity across a given molar mass range. These calibration curves were compared to experimentally obtained curves and, in most cases, excellent agreement was found. The dramatic enhancement in selectivity that can be obtained with iLC in comparison to size-exclusion chromatography (SEC) was illustrated by measuring matrix-assisted laser desorption ionisation (MALDI) MS spectra of fractions collected during a gradient-LC separation. In the low-molar mass range, essentially monodisperse fractions were obtained. Calibration curves, predicted by the model and validated experimentally using narrow-dispersity standards and MALDI-MS spectra of fractions, were used to determine the molar mass distribution of some narrowly distributed polystyrene samples. Molar mass distributions for such standards were found to be somewhat lower than the values reported by the manufacturers. The results also deviated from those obtained by MALDI-MS. PMID- 15782969 TI - Effect of injected sample amount on the shape of chromatographic peaks under condition of linear partition isotherm. AB - In a previous paper a model function was tested in order to approximate the peak shape obtained on non-polar column by injecting different compounds. The simulation of the symmetrical or non-symmetrical shape of gas chromatographic peaks was satisfactory. In this paper, the influence of the amount of injected substance was investigated at different values of inlet pressure and carrier gas velocity, in order to evaluate the relative contribution to the total peak area and shape of the symmetrical distribution due to partition phenomena and of the non-symmetrical and tailing distribution due to adsorption-desorption kinetics. The effect of the molecular mass and of the chain length of compounds belonging to the homologous series of 1-alcohols and n-alkanes on the adsorption phenomena was evaluated. PMID- 15782970 TI - Retention-time database of 126 polybrominated diphenyl ether congeners and two bromkal technical mixtures on seven capillary gas chromatographic columns. AB - The elution order of 126 polybrominated diphenyl ethers (PBDE) was determined for seven different gas chromatographic (GC) stationary phases. The resulting database facilitates selection of the most suitable GC columns for developing a quantitative, congener-specific BDE analysis and the testing of retention prediction algorithms based on structure relationships of GC phases and congener substitution patterns. In addition, co-elutions of the principal BDE congeners with other BDEs and/or with other brominated flame retardants were investigated and, as an application, the composition of two Bromkal mixtures (70-5DE and 79 8DE) was studied. PMID- 15782971 TI - Determination of nonylphenol and octylphenol in paper by microwave-assisted extraction coupled to headspace solid-phase microextraction and gas chromatography-mass spectrometry. AB - A novel and simple method for the determination of active endocrine disrupter compounds (octylphenol OP, and nonylphenol NP) in paper using microwave-assisted extraction (MAE) and headspace solid-phase microextraction, coupled with gas chromatography-mass spectrometry has been developed. Parameters affecting the efficiency in the MAE process such as exposure time and extraction solvent were studied in order to determine operating conditions. The optimised method was linear over the range studied (1.25-125 microg kg(-1) for OP and 9.50-950 microg kg(-1) for NP) and showed good level of precision, with a RSD lower than 10% and detection limits at 0.10 and 4.56 microg kg(-1) for OP and NP, respectively. The results obtained from six different types of paper revealed the presence of the target compounds in all samples analysed, at levels ranging between 3 and 211 microg kg(-1). PMID- 15782972 TI - Dimethyl carbonate as a novel methylating reagent for fatty acids in analytical pyrolysis. AB - Dimethyl carbonate (DMC) was investigated as a mild, harmless and odorless reagent for pyrolytic methylation of fatty acids. Soybean oil was selected as test material for its high content of (poly)unsaturated fatty acids. Pyrolyses were performed at 500, 700 and 900 degrees C by means of a heated platinum filament pyrolyser on-line and off-line to the GC-MS apparatus. Methyl esters of palmitic, linoleic, oleic and stearic acid were formed as prominent products from off-line pyrolysis of soybean oil in the presence of DMC and zeolite 13X. Fatty acid methyl esters (FAMEs) were not observed at important levels in the absence of zeolite, while on-line Py-GC-MS experiments resulted principally in the formation of free fatty acids and hydrocarbons. The FAME profiles obtained from the DMC/zeolite off-line pyrolysis were compared to those resulting from tetramethylammonium hydroxide (TMAH) thermochemolysis and BF3-methanol procedure. The observed differences between pyrolysis and methanolysis methods were principally attributed to the thermal degradation of unsaturated fatty acids. The effectiveness of the DMC/zeolite pyrolytic methylation was further demonstrated by the analysis of tripalmitine and soybean seeds. PMID- 15782973 TI - Analytical characterization of diterpenoid resins present in pictorial varnishes using pyrolysis-gas chromatography-mass spectrometry with on line trimethylsilylation. AB - A procedure based on the technique of the pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) has been applied, in this work, in order to determine the composition of diterpenoid resin employed in art works. The method is based on the on line derivatization of these resins using hexamethyldisilazane (HMDS). Results obtained were compared with those previously reported in literature from Venice turpentine, Strasbourg turpentine, colophony, sandarac and Manila copal using this same method and with those others from in situ thermally assisted hydrolysis and methylation with tetramethylammonium hydroxide (TMAH). Canada balsam, copper resinate and Copaiba balsam have been also analyzed extending the scope of this method in the field of the analysis of artwork materials. Several non-reported trimethylsilylated derivatives of compounds present in the diterpenoid resins have been identified. An improvement in sensitivity has been obtained by using HMDS as derivatizing reagent, together with a better resolution of the most representative peaks. Additionally, this method reduces the number of pyrolytic fragmentation, recombination, dehydration and isomerization products formed during the pyrolysis process and, in consequence, more simplified chromatograms are obtained. Finally, the reported procedure has been successfully used for characterizing two diterpenoid resin-based varnishes present in the canvas painting "The Betrothal of the Virgin" (Anonymous, 17th century) which is included in the pictorial collection of Saint Joseph Church in Taormina (Italy) and the Magdalena Tryptich (Master of Alzira, 16th century, Valencia, Spain). PMID- 15782974 TI - Determination of pharmaceutical residues in waters by solid-phase extraction and large-volume on-line derivatization with gas chromatography-mass spectrometry. AB - This work presents a modified method to analyze selected pharmaceutical residues (clofibric acid, ibuprofen, carbamazepine, naproxen, ketoprofen and diclofenac) in water samples. Various solid-phase extraction cartridges were investigated. The newly developed Oasis HLB (polystyrene-divinylbenzene-N-vinyl pyrrolidone terpolymer) solid-phase extraction (SPE) cartridge provides the optimal sample extraction results. The analytes were then identified and quantitatively determined by gas chromatography-mass spectrometry (GC-MS) via on-line derivatization in the injection-port using a large-volume (10 microl) sample injection with tetrabutylammonium (TBA) salts. This injection-port derivatization technique provides sensitivity, fast and reproducible results for pharmaceutical residues analysis. Mass spectra of butylated derivatives and tentative fragmentation profiles are proposed. Molecular ions and some characteristic ions were used as the quantitation ions to obtain maximum detection sensitivity and specificity. The quantitation limits of these compounds ranged from 1.0 to 8.0 ng/l in 500 ml tap water samples. Recovery of these residues in spiked various water samples ranged from 50 to 108% while RSD ranged from 1 to 10%. The selected analytes were detected in concentrations of 30 to 420 ng/l in wastewater treatment plant effluent and river water samples. PMID- 15782975 TI - Preparation of novel solid-phase microextraction fibers by sol-gel technology for headspace solid-phase microextraction-gas chromatographic analysis of aroma compounds in beer. AB - 3-(Trimethoxysilyl)propyl methacrylate (TMSPMA) was first used as precursor as well as selective stationary phase to prepare the sol-gel-derived TMSPMA-hydroxyl terminated silicone oil (TMSPMA-OH-TSO) solid-phase mircroextraction (SPME) fibers for the analysis of aroma compounds in beer. TMSPMA-OH-TSO was a medium polarity coating, and was found to be very effective in carrying out simultaneous extraction of both polar alcohols and fatty acids and nonpolar esters in beer. The extraction temperature, extraction time, and ionic strength of the sample matrix were modified to allow for maximium sorption of the analytes onto the fiber. Desorption temperature and time were optimized to avoid the carryover effects. To check the matrix effects, several different matrices, including distilled water, 4% ethanol/water (v/v) solution, a concentrated synthetic beer, a "volatile-free" beer and a real beer were investigated. Matrix effects were compensated for by using 4-methyl-2-pentanol as internal standard and selecting the "volatile-free" beer as working standard. The method proposed in this study showed satisfactory linearity, precision and detection limits and accuracy. The established headspace SPME-gas chromatography (GC) method was then used for determination of volatile compounds in four beer varieties. The recoveries obtained ranged from 92.8 to 105.8%. The relative standard deviations (RSD, n = 5) for all analytes were below 10%. The major aroma contributing substances of each variety were identified via aroma indexes. PMID- 15782976 TI - Capillary electrophoresis enhanced by automatic two-way background correction using cubic smoothing splines and multivariate data analysis applied to the characterisation of mixtures of surfactants. AB - Mixtures of the surfactant classes coconut diethanolamide, cocamido propyl betaine and alkylbenzene sulfonate were separated by capillary electrophoresis in several media containing organic solvents and anionic solvophobic agents. Good resolution between both the surfactant classes and the homologues within the classes was achieved in a BGE containing 80 mM borate buffer of pH 8.5, 20% n propanol and 40 mM sodium deoxycholate. Full resolution, assistance in peak assignment to the classes (including the recognition of solutes not belonging to the classes), and improvement of the signal-to-noise ratio was achieved by multivariate data analysis of the time-wavelength electropherograms. Cubic smoothing splines were used to develop an algorithm capable of automatically modelling the two-way background, which increased the sensitivity and reliability of the multivariate analysis of the corrected signal. The exclusion of significant signals from the background model was guaranteed by the conservativeness of the criteria used and the safeguards adopted all along the point selection process, where the CSS algorithm supported the addition of new points to the initially reduced background sample. Efficient background modelling made the application of multivariate deconvolution within extensive time windows possible. This increased the probability of finding quality spectra for each solute class by orthogonal projection approach. The concentration profiles of the classes were improved by subsequent application of alternating least squares. The two-way electropherograms were automatically processed, with minimal supervision by the user, in less than 2 min. The procedure was successfully applied to the identification and quantification of the surfactants in household cleaners. PMID- 15782977 TI - Gas chromatographic-mass spectroscopic determination of benzene in indoor air during the use of biomass fuels in cooking time. AB - A gas chromatography-mass spectroscopic method in electron ionization (EI) mode with MS/MS ion preparation using helium at flow rate 1 ml min(-1) as carrier gas on DB-5 capillary column (30 m x 0.25 mm i.d. film thickness 0.25 microm) has been developed for the determination of benzene in indoor air. The detection limit for benzene was 0.002 microg ml(-1) with S/N: 4 (S: 66, N: 14). The benzene concentration for cooks during cooking time in indoor kitchen using dung fuel was 114.1 microg m(-3) while it was 6.6 microg m(-3) for open type kitchen. The benzene concentration was significantly higher (p < 0.01) in indoor kitchen with respect to open type kitchen using dung fuels. The wood fuel produces 36.5 microg m(-3) of benzene in indoor kitchen. The concentration of benzene in indoor kitchen using wood fuel was significantly (p < 0.01) lower in comparison to dung fuel. This method may be helpful for environmental analytical chemist dealing with GC-MS in confirmation and quantification of benzene in environmental samples with health risk exposure assessment. PMID- 15782978 TI - Validated, non-destructive and environmentally friendly determination of cocaine in euro bank notes. AB - A non-destructive, fast and environmentally friendly procedure has been developed for cocaine determination in euro bank notes. Cocaine was extracted with 15 ml methanol by vortex agitation during 5 min. The extract was evaporated and reconstituted in 0.5 ml methanol. GC-MS-MS analysis was performed using as precursor ion m/z 182.2, with an excitation energy voltage of 1.60 eV, being the product ions measured m/z 150.2 and 82.0. A limit of detection of 0.15 ng per note and a repeatability of 6%, established from the relative standard deviation, of a 1 ng ml(-1) level, were achieved. Recoveries of 101+/-2 and 98+/-3% were obtained for samples spiked with 100 and 10 microg respectively. Results show that all the euro bank notes measured (16 samples) were contaminated with cocaine in the range between 1.25 and 889 microg. Two different contamination levels, high level (150-889 microg) and low one (1.25-77 microg) were found and it could be related with the direct or indirect contact with the drug. PMID- 15782979 TI - Application of capillary zone electrophoresis to the analysis and to a stability study of nitrite and nitrate in saliva. AB - The applicability of capillary zone electrophoresis for the determinations of nitrite and nitrate was studied. Using direct UV detection the limit of detection values of the analytes were 0.14 and 0.21 microg/mL, respectively. The developed method was found to be useful to directly determine nitrite, nitrate and thiocyanate in saliva. It was found that adjusting the pH of the sample to 11 and storing the saliva at 4 degrees C was adequate to make constant the nitrite/nitrate ratio in saliva samples at least 7 days. PMID- 15782980 TI - Long-term strict monitoring of plasma ghrelin and other serological markers of gastric diseases after Helicobacter pylori eradication. AB - We report on a case of chronic atrophic gastritis in which the serological markers of gastric diseases were strictly monitored for 2 years after successful Helicobacter pylori (H. pylori) eradication. A 31-year-old man with upper abdominal pain was diagnosed as having H. pylori infection. Laboratory examination revealed low serum levels of pepsinogen (PG) I, low PG I/II ratio, and low plasma levels of ghrelin. Upper gastrointestinal endoscopy revealed severe corpus-dominant atrophic gastritis. H. pylori eradication therapy was performed. Successful eradication was confirmed three months later by the 13C urea breath test. Decreased serum PG II levels and an increased serum PG I/II ratio were detected a week after completion of the eradication therapy. The serum anti-H. pylori IgG titer decreased to less than 75% of the baseline level by 24 weeks after completion of the eradication therapy. On the other hand, the plasma levels of total and active ghrelin showed no marked changes after successful eradication therapy. This is the first report of long-term follow-up of changes of the plasma ghrelin levels after H. pylori eradication therapy, the observations suggesting that reduction of plasma ghrelin levels cannot be achieved merely by H. pylori eradication, without resolution of the gastric atrophy. PMID- 15782981 TI - Gallbladder carcinoma in Jordan. AB - BACKGROUND/AIMS: Despite advances in diagnostic and therapeutic modalities, the prognosis of gallbladder carcinoma is still poor. The purpose of this study is to look for peculiarities of primary gallbladder carcinoma in Jordan regarding its incidence, clinical and pathological aspects. METHODOLOGY: A retrospective study over a period of 19 years comprising 66 patients was reviewed; only patients treated primarily in our hospital were included. There were 40 females and 26 males (ratio of 3:2). RESULTS: The main presenting symptom was abdominal pain in 54 patients (82%); correct preoperative diagnosis was made in only 20% of cases. Cholecystectomy alone was the most commonly performed operation (32 cases), cholecystectomy in combination with biliary bypass and/or hepatic resection (16 cases), biopsy of the gallbladder was performed in 12 cases and bypass with T tube drainage in 6 cases. Postoperative morbidity and mortality were encountered in 48% and 18% respectively. CONCLUSIONS: The incidence and clinicopathological aspects of gallbladder cancer in Jordan seem to resemble that in the west, albeit with a lower age incidence. Diagnosis remains difficult and the outcome is still poor in the majority of cases. PMID- 15782982 TI - Implications of arterial anatomy in patients with cancer of the periampullary region. AB - BACKGROUND/AIMS: Preoperative information on arterial anatomy in the peripancreatic and hepatic areas is valuable to any surgeon performing pancreatoduodenectomy. METHODOLOGY: Between 1994 and 1998, 49 patients with periampullary cancer (31 distal bile duct and 18 ampullary tumors) underwent visceral angiography and radical pancreatoduodenectomy with lymphadenectomy. Surgically "significant" arterial variations and their effects on operative management and results were examined retrospectively. RESULTS: Arterial variations were found in 18 patients (37%); 15 (31%) were "significant" and 3 (6%) were "nonsignificant." All 15 patients with "significant" variants required specific type of various preservations of the hepatic arterial system. Intraoperative blood loss, transfused blood units, and operation time were greater in patients with "significant" variations than in patients without (P<0.05). Histopathologic diagnosis, tumor staging, morbidity, and mortality did not differ between the 2 groups. The 5-year survival was 33% for patients with "significant" variations and 63% for patients without (P<0.05). CONCLUSIONS: Information on arterial anatomy in the peripancreatic and hepatic areas is necessary for preoperative evaluation in patients requiring radical pancreatoduodenectomy. Presence of "significant" arterial variations may be considered as one of the negative prognostic factors in patients with periampullary cancer. PMID- 15782983 TI - Complete resection of choledochal cyst with Roux-en-Y derivation vs. cyst enterostomy as standard treatment of cystic disease of the biliary tract in the adult patient. AB - BACKGROUND/AIMS: Cystic disease of the biliary tract (CDBT) is characterized by varying degrees of dilatation of the biliary system with high morbidity and mortality in which the surgical management is the corner stone of treatment. The cyst-enterostomies (CE) temporarily solve the obstruction to the biliary flow but have a high long-term morbidity. Complete resection of affected bile ducts with Roux-en-Y derivation (CRR-en-Y) is a good procedure with low mortality and complications. The purpose of the study was to analyze the outcome of CRR-en-Y versus CE in the treatment of CDBS in adult patients from 1970 to 2002. METHODOLOGY: Patients who underwent surgical treatment were divided for their analysis into two groups: Group I: CRR-en-Y and Group II: CE. Following features were compared: demography, clinical picture, postoperative morbimortality, outcome and survival. RESULTS: Thirty-four adult patients were analyzed. There were 82% (28) females and 18% (6) males. The age average was 33.58 years (13-84). Seventy percent (30) were "choledochal cyst". Eighty percent (27) were surgically handled: 52% (14) with CRR-en-Y (Group I) versus 58% (13) with CE (Group II). Both groups were comparable. Without operative mortality and low postoperative morbidity in both, CE had more long-term complications: In this group 70% (9) were readmitted: and 7 underwent reoperation. Mean follow-up was 35 months (6 132) versus 152 months (12-408) respectively. CONCLUSIONS: CRR-en-Y is the standard treatment of CDBS in the adult patient. PMID- 15782984 TI - Laparoscopic cholecystectomy for acute cholecystitis in the elderly: a retrospective study. AB - BACKGROUND/AIMS: Laparoscopic cholecystectomy for acute cholecystitis in the elderly remains a subject almost unstudied. A first study performed at Brugmann hospital broached the subject, but was rather unsatisfying. In fact, it showed that, except for a shorter hospital stay, laparoscopic cholecystectomy didn't have any advantages over open cholecystectomy. In view of the perfection in surgical technique and of the rapidity of management, we have analyzed our new results and compared them to the ones of our former study. METHODOLOGY: In this current study we included patients aged over 75 years and with histologically proven acute cholecystitis. As such, we retrospectively studied 20 patients undergoing cholecystectomy from 1997 to 2002. Sixteen patients (80%) underwent laparoscopic cholecystectomy whereas 4 (20%) were considered unable to withstand the laparoscopic approach because of hemodynamic instability, they underwent open cholecystectomy. RESULTS: Age, APACHE-II scoring, white blood cell counts and CRP were more important in the laparotomy group, but there was no difference in terms of local and general complication or mortality rates between the two groups. CONCLUSIONS: Comparing the results with those of the first study (1991-1997), we established that the laparoscopic approach is very well indicated in the geriatric population. Also, the laparoscopy has been applied more appropriately, seen that our conversion rate dropped to zero. Still, open cholecystectomy proves to be indicated in critically ill patients. PMID- 15782985 TI - An investigation of primary malignancies associated with ampullary carcinoma. AB - BACKGROUND/AIMS: As ampullary carcinoma has a favorable prognosis, associated malignancies have potential prognostic significance in these patients. This study focused on the incidence and characteristics of preexisting, coexisting and subsequent malignancies in patients with ampullary carcinoma. METHODOLOGY: Sixty four cases of ampullary carcinoma were retrospectively identified. Fifty-four patients underwent surgical resection, while in the other ten, histological diagnosis was reached after biopsy and/or autopsy. Other primary malignancies associated with ampullary carcinoma, occurring in the prediagnostic or postdiagnostic period, were investigated. The mean postdiagnostic follow-up period was 3.9 years (range, 0-23 years). RESULTS: Other malignancies occurred in 17 patients (27%), being found before (n=9), at (n=7), or after (n=5) diagnosis of ampullary carcinoma. The total number of associated lesions was 23, including one case of quadruple and four cases of triple carcinoma. The major associated malignancies were colonic carcinoma (n=9), gastric carcinoma (n=5), and uterine carcinoma (n=3). Twenty-one lesions were treated surgically or endoscopically. One patient died from synchronous extrahepatic bile duct carcinoma and one from leukemia, and one died from subsequent urinary bladder carcinoma. Development of other malignancies was related to age but not to gender, family history, adjuvant chemo/radiotherapy, or tumor pathology. CONCLUSIONS: Ampullary carcinoma is associated with a high incidence of other malignancies, particularly colonic and gastric carcinomas. The possibility of associated malignancies should therefore be considered in preoperative screening and postoperative follow-up of patients with ampullary carcinoma. PMID- 15782986 TI - Hydrodynamics-based gene delivery to the liver by bile duct injection of plasmid DNA--the impact of lasting biliary obstruction and injection volume. AB - BACKGROUND/AIMS: Increased biliary pressure may mediate an opening of intrahepatic tight junctions. The biliary pressure is related to the duration of biliary obstruction and the injected volume. Their effects on the efficiency of gene delivery to the liver were studied. METHODOLOGY: The luciferase gene with a volume of 0.5, 1.0, 1.5 or 2.0mL was injected into the bile duct of Sprague Dawley rats. After the injection, the bile ducts were occluded in some of the rats, while left to drain in the others. RESULTS: The activity of luciferase was the highest in rats that received 2.0mL of DNA injection and biliary occlusion, while the lowest in the rats that received 0.5mL of DNA only. CONCLUSIONS: The bile duct can be an efficient route for gene delivery to the liver depending on the volume of injected DNA. Biliary occlusion can enhance the expression of genes but it may induce a certain degree of liver injury, which should be taken into concern. PMID- 15782987 TI - Classification of choledochocele. AB - BACKGROUND/AIMS: Choledochocele is a rare abnormality of cystic or diverticular dilatation of the terminal intramural portion of the common bile duct. Because its anatomic form does not fit the criteria for pancreaticobiliary maljunction, reflux of bile or pancreatic juice has not been discussed in relation to choledochocele. Considering the flow of bile and pancreatic juice, the authors developed a new classification of choledochocele, and investigated associated pancreaticobiliary diseases. METHODOLOGY: We reviewed 82 cases of choledochocele, and classified them into three groups on cholangiopancreatographic findings; Type A (n=27): cystic dilatation of the common channel, Type B (n=31): cystic dilatation of the intramural bile duct forming the common channel, Type C (n=24): cystic dilatation of the intramural bile duct opening into the duodenum separately from the pancreatic duct. RESULTS: Biliary carcinoma was associated with 2 cases of Type A and 1 case of Type B choledochocele. Acute pancreatitis occurred in 10 Type A and 6 Type B choledochoceles. Biliary pancreatic reflux was reported in 4 cases of Type A and 1 case of Type B. Marked elevation of amylase in the bile was reported in 5 Type A and 5 Type B choledochoceles. CONCLUSIONS: In choledochoceles forming a common channel (Type A and B), two-way regurgitation may occur, and induce acute pancreatitis and biliary carcinoma. This classification of choledochocele is useful for analysis of pathophysiology and determination of therapeutic strategy. PMID- 15782988 TI - A proposed inflammation grading system for laparoscopic cholecystectomy. AB - BACKGROUND/AIMS: Currently, there is no comprehensive method to classify the inflammatory status of the GB and Calot's triangle. METHODOLOGY: From 1996, we performed LC on 111 patients with symptomatic gallbladder diseases, irrespective of onset, using an abdominal wall-lifting method. Thirty-six patients had acute cholecystitis, while thirty had chronic cholecystitis. Severe inflammation prompted the author to design a grading system to record inflammatory status and to evaluate the feasibility of LC prospectively. GB with none - mild, moderate, severe wall thickening, and Calot's triangle involvement with mild - moderate, and severe inflammation were classified as Grade I - V, respectively. RESULTS: Forty-six cases had severe inflammation greater than grade III, however, neither CBD injury nor mortality was experienced. Complications were minimal. Of ten conversion cases (9.0%), eight were grade V, one grade III, and one grade I. Poor laparoscopic view caused by incomplete muscle relaxation accounted for four of the conversions. CONCLUSIONS: Our grading system is useful in evaluating feasibility of difficult gallbladders, helping to decide conversion during LC, and as a tool to record or elucidate the chronological inflammatory changes of various GB disorders. PMID- 15782989 TI - Beneficial effects of ERCP and papillotomy in predicted severe biliary pancreatitis. AB - BACKGROUND/AIMS: Extensive circumstantial evidence indicates that patients with a predicted severe attack of acute biliary pancreatitis (ABP) should undergo an endoscopic retrograde cholangiography with papillotomy (ERC/PT). However, in clinical practice this procedure is not always performed. This study was conducted to compare outcome in patients with and without ERC/PT. METHODOLOGY: Thirty-five of 80 patients admitted with ABP had a predicted severe attack (three or more Ranson criteria). Only in 24 of these 35 patients was an ERC/PT performed. RESULTS: In the ERC/PT group, significantly less pancreatic necrosis (8 vs. 64%, p<0.001) occurred, hospital stay was shorter (median 22 +/- 5 vs. 51 +/- 19 days, P=0.08) and mortality was lower (8 vs. 36%, P=0.01). Twenty-three patients (66%) underwent cholecystectomy after a median period of 10 weeks (range 0-26 weeks) after discharge. During the waiting period, in the ERC/PT group, two patients developed acute cholecystitis whereas recurrent ABP and common bile duct stones occurred in one patient each. CONCLUSIONS: In patients with a predicted severe attack of ABP, performing ERC/PT is associated with less morbidity and lower mortality. PMID- 15782990 TI - Prognostic implications of preserved bile duct confluence after iatrogenic injury. AB - BACKGROUND/AIMS: Biliary reconstruction is performed according to the level of the injury. A comparative study between patients in whom the biliary junction was preserved and another group where the biliary junction was not preserved was done. METHODOLOGY: A retrospective review of the biliary reconstructions performed at our institution after iatrogenic lesions between 1990-2002 was done. Postoperative outcome, functional status of the anastomosis, recurrent cholangitis, need for radiological instrumentation and/or reoperation were analyzed. RESULTS: We reviewed 204 cases, 130 cases had a preserved biliary junction while in 74 the injury included the junction. All patients were treated with a Roux-en-Y hepatojejunostomy. In the first group, 4% required reoperation, 4% underwent radiological percutaneous instrumentation, 8% had anastomotic dysfunction and 4% cholangitis. In the second group, 24% needed reoperation and 80% radiological instrumentation. Anastomotic dysfunction was observed in 64% and cholangitis in 55%. It is important to note that 52 of the 74 cases in the second group had a history of more than two reconstruction attempts. CONCLUSIONS: When the biliary junction is preserved after a iatrogenic injury we found a significantly better outcome. The results of biliary reconstruction in this type of patient are better long-term compared to those where the junction was not preserved, evidenced by a lower reoperation and radiological instrumentation rate. PMID- 15782991 TI - Effect of preoperative endoscopic decompression on malignant biliary obstruction and postoperative infection. AB - BACKGROUND/AIMS: The role of preoperative biliary drainage as an adjunct in patients undergoing surgical resection for malignant biliary obstruction is controversial. The objective of the present study was to evaluate the effects of preoperative endoscopic biliary drainage and its possible association with postoperative infection in patients presenting neoplastic obstructive jaundice. METHODOLOGY: The medical records of 53 patients presenting periampullary neoplasia were reviewed. In all of these patients, bile was obtained intraoperatively by puncture. Among the 53 patients, 14 had been submitted to preoperative endoscopic drainage (Group I) and 39 had undergone surgery without decompression (Group II). For statistical analysis, the level of significance was set at 5%. RESULTS: Bile culture was positive in 13 (92.8%) of the patients in Group I, a significantly higher incidence than that observed in group II, in which 16 (41%) of the patients presented positive cultures (p=0.001). There was no significant difference in general postoperative morbidity between groups (64.2% for group I vs. 46% for group II) or mortality (0% vs. 7.6%, respectively). When infective complications (cholangitis, pneumonia, wall infection) were analyzed separately, a higher incidence, although without significance, was found in Group I than in Group II (50% vs. 28.2%, respectively; p=0.1913). CONCLUSIONS: The presence of an endoscopic biliary drain provokes bacterial colonization, possibly due to the combination of residual cholestasis and duodenal reflux to the bile duct, raising concerns about the possible appearance of infective complications during the postoperative period. PMID- 15782992 TI - Gallbladder reefing for cholelithiasis as a complication of colon cancer. AB - In an 83-year-old woman presenting with gallstones and a cancer in the sigmoid colon, resection was performed through a median incision, after which the wound was extended, the stones were crushed, and the gallbladder was infolded and sutured (reefed). Even in elderly patients, some treatment for bile stones should be done at abdominal section to avoid future cholecystitis or complications. Reefing is a useful technique that can be done easily from a comparatively narrow field of view. PMID- 15782993 TI - Renal failure in obstructive jaundice. AB - BACKGROUND/AIMS: The optimal preoperative management of patients with acute obstructive jaundice is still a matter of debate. Reduction in effective plasma volume and systemic endotoxemia are major consequences both in experimental acute bile duct ligation and in patients with acute obstructive jaundice (AOJ). The objective of this study is to show the necessity for adequate preoperative hydration and lactulose administration for the prevention of postoperative renal dysfunction in patients with AOJ. METHODOLOGY: Fifty-five patients (35 male, 20 female) with a mean age of 62 years were entered into the study. There were 23 benign and 32 malignant strictures in this group. All patients received oral lactulose (30-45mL per day) and IV cefazoline (3-4g/day) before surgery. Fluid and electrolyte balance was precisely maintained throughout the hospitalization via daily body weight calculations. High-risk elderly patients with left ventricular systolic dysfunction were assessed by echocardiography and therapeutic measures were undertaken. Renal function was assessed by creatinine clearance (ClCr) measurements and impairment of renal function was defined as a >20% fall in ClCr value post-surgery. RESULTS: Mean preoperative bilirubin level was 11.05mg/dL (range: 3.45-27.0mg/dL). None of the patients developed postoperative renal failure. The difference between pre- and postoperative ClCr value (104.02mL/min vs. 101.0mL/min respectively) was not significant (t=0.698, P=0.489). One patient with mild renal function impairment before surgery (ClCr=45.4mL/min) successfully recovered with 60% rise in creatinine clearance (ClCr=78.1mL/min) after the operation. Three patients with normal functioning kidneys died within 30 days of operation. The principle cause of death was carcinomatosis and pulmonary embolus in two and intraoperative hemorrhagic shock in one patient. CONCLUSIONS: These results further substantiated the importance of adequate preoperative hydration and endotoxin inactivation in terms of acute renal failure prophylaxis in patients with AOJ. This regimen obviates the afferent events in obstructive jaundice and provides prevention of acute renal failure even in high-risk elderly patients. PMID- 15782994 TI - Disruption of pRb-p16INK4 pathway: a common event in ampullary carcinogenesis. AB - BACKGROUND/AIMS: Aberrations of the pRb (Retinoblastoma gene protein)-p16INK4 pathway play a critical role in carcinogenesis. Our objective is to evaluate its role in tumorigenesis and the development of ampullary cancer. METHODOLOGY: We examined expression status of p16INK4 protein and pRb immunohistochemically and assessed their possible prognostic relevance in 36 ampullary cancers. RESULTS: Thirty-four specimens (94.4%) exhibited alteration of p16INK4 and/or pRb expression, with 63.9% (23/36) of cancers showing p16INK4 negative expression and 94.4% (34/36) pRb abnormal expression. p16INK4 protein negative expression correlated significantly with tumor progression features such as advanced tumor stages (p=0.0291), lymph node metastasis (p=0.005), pancreas invasion (p=0.0002) and duodenum invasion (p=0.0101). Cases with both p16RNK4 protein negative expression and pRb overexpression showed poorer differentiation, more invasive growth (p=0.0425), higher level tumor stages (p=0.0079) and more frequent pancreas invasion (p=0.0024), compared with the others. p16INK4 protein expression showed no relationship with pRb expression (p=0.2199). No association was found in pRb expression status compared with any clinicopathological parameters analyzed. CONCLUSIONS: The disruption of the pRb-p16NK4 pathway plays an important role in ampullary carcinogenesis, the absence of p16INK4 protein expression might be involved in ampullary tumor progression. PMID- 15782995 TI - Possible endocrine control by follistatin 315 during liver regeneration based on changes in the activin receptor after a partial hepatectomy in rats. AB - BACKGROUND/AIMS: Activin A is an autocrine inhibitor of cell growth in the liver. The biological activity of activin A is mediated by a heteromeric receptor complex. Follistatin (FS) binds to activin and inhibits its biological effects, and acts as a negative regulator of muscle cells. The role of activin receptors during liver regeneration following a hepatectomy has not been fully assessed. This study investigates the mechanism underlying how activin receptors regulate hepatocyte growth, and the effects of intravenous administration of FS during liver regeneration. METHODOLOGY: The expression of both activins and activin receptors in the liver after a 70% partial hepatectomy (HT) was assessed by RT PCR and immunohistochemistry. FS 315 or 288 was infused for different periods of time based on changes in hepatocyte activin receptor expression after HT. RESULTS: Activin receptor expression peaked between 48 and 72 hours after HT. 72 hours after HT, an injection of FS 315 resulted in a more potent stimulation of DNA synthesis and produced a greater increase in body weight compared with the control rats. CONCLUSIONS: The expression of activin receptors after peak DNA synthesis might be a key component in the downregulation of DNA synthesis. Intravenous administration of FS 315 might promote liver regeneration and have anabolic actions. PMID- 15782996 TI - Histochemical expression of sialoglycoconjugates in carcinoma of the papilla of Vater. AB - BACKGROUND/AIMS: Cell surface glycosylation changes during oncogenesis and is thought to correlate with the malignant potential of cancers. To investigate the role of sialylation in carcinoma of the papilla of Vater, histochemical analyses were performed using sialic acid-binding lectins, Maackia amurensis leukoagglutinin and Sambucus nigra agglutinin. METHODOLOGY: Thirty-six papillary carcinoma tissues and 8 normal papillary tissues were subjected to lectin histochemical staining. The relationship between lectin staining characteristics and clinicopathological parameters was statistically analyzed. RESULTS: Epithelial cells of glands in normal tissues were stained with Sambucus nigra agglutinin but not with Maackia amurensis leukoagglutinin. Of 36 papillary carcinoma tissues, 20 showed Maackia amurensis leukoagglutinin-positive staining in cancer cells, whereas all 36 were Sambucus nigra agglutinin-positive in cancer cells. According to Maackia amurensis leukoagglutinin-staining characteristics, the 36 cases were divided into two groups: positive (n=20) and negative (n=16). Statistical analysis indicated that positive staining in cancer cells was frequent in cases of lymph node metastasis (p=0.001) and in cases classified > or = T2 by TNM classification (p=0.036). CONCLUSIONS: Aberrant expression of sialoglycoconjugates recognized by Maackia amurensis leukoagglutinin might participate in lymph node metastasis and in an advanced stage of invasion in carcinoma of the papilla of Vater. PMID- 15782997 TI - Three-dimensional power Doppler ultrasonography for hepatocellular carcinoma: a comparison with angiography? AB - BACKGROUND/AIMS: To assess clinical usefulness of three-dimensional power Doppler ultrasonography for the diagnosis of hepatocellular carcinoma. METHODOLOGY: Fifty two hepatocellular carcinoma nodules (median 31mm in diameter, range 8 to 94), histologically proven afterwards, in 33 patients were examined by three dimensional power Doppler ultrasonography (ATL-HDI 5000 with a 5-MHz convex transducer) for tumor vascularity. We classified tumor Doppler signals into four types; Type 1: spotty signals in the tumor, Type 2: signals surrounding the tumor, Type 3: Type 3 with visualized penetrating arteries, Type 4: Type 3 with visualized drainage vein. Types 2-4 were considered specific to hepatocellular carcinoma, and compared with findings on digital subtraction angiography. RESULTS: Definite diagnosis of hepatocellular carcinoma was obtained in 29 of 52 nodules (56%) with three-dimensional ultrasonography (5 with Type 2, 19 with Type 3, and 5 with Type 4) while all nodules revealed tumor stain on angiography. 23 nodules showed only Type 1 signals, which were not specific to hepatocellular carcinoma. These nodules included small nodules less than 2cm in diameter, located more than 5cm from body surface, and those in the subphrenic portion of the left lobe. CONCLUSIONS: Three-dimensional power Doppler ultrasonography provides definite diagnosis of hepatocellular carcinoma in a real-time, non invasive manner under certain conditions. PMID- 15782998 TI - Hyperinsulinemia in patients with colorectal cancer. AB - BACKGROUND/AIMS: It has been reported that non-insulin dependent diabetes mellitus (NIDDM) is one of the risk factors for colorectal cancer. Usually, in the pre-NIDDM state, hyperinsulinemia is seen for 5 to 8 years. Insulin is the growth factor of epithelial and cancer cells of colon and rectum. In this study, we evaluate glucose tolerance in the patients with colorectal cancer who were never diagnosed with DM. METHODOLOGY: We studied 82 patients with colon cancer who were never diagnosed with DM. 75-g glucose tolerance test (75g GTT) was performed and we measured serum glucose (BS) and insulin (IRI) levels, and we defined them as normal glucose tolerance (NGT), impaired GT (IGT), and DM. We also defined hyperinsulinemia as highest IRI levels over 100mU/mL at 75g GTT. RESULTS: Serum glucose and insulin levels were higher in the patients with colorectal cancer than in healthy controls. In 82 colorectal cancer patients, 39 were IGT and 5 were DM. All DM patients also had hyperinsulinemia. Only 14 patients (17%) had NGT and normal IRI levels. CONCLUSIONS: Our findings suggest that hyperinsulinemia is occasionally seen in patients with colorectal cancer. Hyperinsulinemia may be one of the causes of colorectal cancer and we have to control hyperinsulinemia to prevent recurrence of colorectal cancer even after curative resection. PMID- 15783000 TI - Microsatellite instability in poorly differentiated colorectal adenocarcinoma, particularly in relation to two subtypes. AB - BACKGROUND/AIMS: There have been a few reports indicating the characteristics of poorly differentiated colorectal adenocarcinoma by dividing it into subtypes. However, we have not found any reports describing microsatellite instability (MSI) of each subtype. In this study, we elucidated the clinicopathological features of subtypes in the poorly differentiated adenocarcinoma, especially the relationship between MSI and each subtype. METHODOLOGY: The present study included 28 cases with poorly differentiated adenocarcinoma. The 28 cases were classified into two groups; the solid group and the non-solid group. For each group, the clinicopathological aspects and MSI were examined. RESULTS: No significant differences were noted between the solid group and the non-solid group in terms of clinicopathological findings excluding male/female ratio. The 5 year survival rate of the solid group (38.5%) was significantly higher than that of the non-solid group (0.00%) (p=0.0013). The ratio of cases with MSI-H in the solid group (80.0%=12/15) was significantly higher than that of the non-solid group (30.8%=4/13) (p=0.0087). CONCLUSIONS: The incidence of MSI as well as the prognosis was different between solid and non-solid type with poorly differentiated colorectal adenocarcinoma. Therefore, we think that poorly differentiated colorectal adenocarcinoma should be classified into two subtypes: solid type and non-solid type when analysis for poorly differentiated adenocarcinoma is performed. PMID- 15782999 TI - The limited use of ultrasound in the detection of abdominal abscesses in patients after colorectal surgery: compared with gallium scan and computed tomography. AB - BACKGROUND/AIMS: Ultrasound has been suggested to be a first-line test for the detection of intra-abdominal abscess (IAA). However, according to our experience, ultrasound seemed not so effective when applied to the patients after colorectal surgery. Computed tomography (CT) and gallium scan have also been suggested as very useful for the detection of IAA. The purpose of this study was to evaluate the efficacy of sonography and gallium scan in diagnosing intra-abdominal abscess after colorectal surgery and to compare these results with CT. METHODOLOGY: Twenty-three patients with suspected IAA after colorectal surgery at our institution were eligible for the study. A positive IAA was verifed only if pus was drained or on postmortem examination. Negative reports of abscesses were verified by adequate outpatient review following hospital discharge that indicated the clinical absence of an abscess. Within 4 days of the clinical suspicion of abscess, all patients were evaluated by three techniques: sonography, CT and gallium scan. We compared the sensitivity, specificity and accuracy of these modalities. RESULTS: Fourteen of the 23 patients (60.9%) were verified as IAA. We found the sensitivity, specificity, and accuracy of CT scan for the detection of LAA in the patients with colorectal surgery to be 92.9%, 100%, and 95.7%, respectively. The sensitivity, specificity and accuracy of gallium scan were 100%, 77.8% and 91.3%, respectively. Ultrasound had a 42.9% sensitivity, 100% specificity, and 65.2% overall accuracy for diagnosing IAA. CONCLUSIONS: Sonography plays a small role in the detection of IAA in the patients after colorectal surgery. CT and gallium scan are more helpful than sonography for the detection of IAA. PMID- 15783001 TI - Minimal number of lymph nodes that need to be examined for adequate staging of colorectal cancer--factors influencing lymph node harvest. AB - BACKGROUND/AIMS: Accurate assessment of lymphatic node status is an essential component in staging of colorectal cancer and determining the need for adjuvant treatment. The risk of understaging nodal status decreases by increasing the number of examined lymph nodes. Several recommendations have been published for the minimum number of lymphatic nodes that need to be harvested for accurate nodal staging with minimal chance of error, ranging from 6 to 17 lymph nodes. However, the number of harvested lymph nodes is very variable, and this variability is poorly understood. The aim of this study was to determine factors associated with the number of retrieved lymph nodes in patients with colorectal cancer. METHODOLOGY: Clinical and histological characteristics of 177 patients operated for colorectal cancer were analyzed. RESULTS: In multiple regression analysis, male gender, better tumor differentiation and greater tumor size, as well as the presence of acute inflammation were determined as significant independent predictors of the increased number of resected and examined lymph nodes. CONCLUSIONS: The determination of the minimum number of lymph nodes required for accurate nodal staging of patients with colorectal cancer needs to be individualized. PMID- 15783002 TI - Surgical results of total pelvic exenteration for locally advanced colorectal adenocarcinoma. AB - BACKGROUND/AIMS: Although total pelvic exenteration (TPE) may lead to local tumor control and improved quality of life in patients with locally advanced colorectal cancer, an adequate understanding of prognostic factors, indications and potential complications associated with these procedures is needed. METHODOLOGY: Records for 15 patients, who underwent TPE for colorectal adenocarcinoma at Oita Prefectural Hospital between January 1983 and November 2001, were reviewed, retrospectively. RESULTS: Ten (66.7%) had positive lymphatic involvement, seven (46.7%) had positive vascular involvement, and three (20%) had positive lymph node metastases. Bladder involvement histologically was evident in eight patients (53.3%). With regard to diagnostic assessment of bladder involvement using CT, the sensitivity and specificity were 83.3% and 60%, respectively. Six of 15 patients (40%) developed complications. Overall local recurrence was observed in 6 (40%) of the 15. The cumulative overall 5-year survival rate of the 15 patients in this study was 54.7%. In the univariate analysis, vascular involvement significantly influenced survival. CONCLUSIONS: TPE appears to be relatively safe and effective for treatment of locally advanced colorectal adenocarcinoma. Vascular involvement was recognized as the only reliable prognostic clinicopathological characteristic. PMID- 15783003 TI - Local recurrence following curative low anterior resection for rectal carcinoma. AB - BACKGROUND/AIMS: Local recurrence is a formidable problem after potentially curative resection for rectal cancer. We attempted to identify possible factors affecting the frequency of local recurrence, focusing on the clearance of the tumor and the margin of resection. METHODOLOGY: The clinical cohort consisted of 66 patients suffering from rectal carcinoma. All patients underwent a low anterior resection with meticulous pelvic preparation and dissection. The proximal and distal margins of the tumor were measured before fixing for permanent sections. RESULTS: Analysis by distance of the tumor from the anal verge revealed that 5 out of 33 patients (15.15%) from the upper rectal group and 7 out of 19 patients (36.8%) from the mid rectal group developed local recurrences (36.8% vs. 15.15% P=0.0369). Analysis by distance of the distal resection margins revealed that 8 out of 12 patients with local recurrence had distal margins less than 2cm, whereas 7 out of 40 patients free of local relapse had margins less than 2cm. Therefore 8 out of 15 patients with distal margins less than 2cm developed local recurrence (53.5%) whereas 4 patients among 37 with margins greater than 2cm developed a local failure (10.8%) (53.5% vs. 10.8% P=4.88E-04). The median survival in all patients of our series was 23 months. The median survival in 52 patients who underwent a potentially curative resection was 42 months. CONCLUSIONS: Tumors located in the upper portion of the rectum presented a minor tendency for local recurrence compared to tumors located in the middle of the rectum. Our study provides strong indications that high rate of local recurrences are probably related to the limited anatomic margins that can be obtained in the pelvis during primary resections. PMID- 15783004 TI - The effect of the herbal medicines dai-kenchu-to and keishi-bukuryo-gan on bowel movement after colorectal surgery. AB - BACKGROUND/AIMS: To evaluate the safety and efficacy of the herbal medicines Dai ken-chu-to and Keish-bukuryo-gan in the patients undergoing colorectal resection. METHODOLOGY: Sixty-six patients undergoing colorectal resection were studied. Patients in the Study group began on 7.5g of Dai-ken-chu-to and 6.0g of Keish bukuryo-gan on the first postoperative day. Control group did not take herbal medicine at all. All of the patients were monitored for vomiting, abdominal distention, length of ileus, tolerance of regular diet, duration of intravenous fluid administration, length of hospitalization, and complications. RESULTS: The time to flatus was 63.1 +/- 22.8 hours and 95.4 +/- 33.0 hours in the Study and Control group, respectively (p<0.05). And the time to tolerance of regular diet was 2.53 +/- 1.93 days and 6.25 +/- 1.50 days in the Study and Control group, respectively (p<0.05). In addition, length of postoperative hospitalization in the patients within the Study group was significantly shorter than those within the Control group (15.7 +/- 4.8 days vs. 20.8 +/- 4.8 days, p<0.05). The incidence of complications including nausea, vomiting, anastomotic leak and wound infection occurred equally in both groups. CONCLUSIONS: Administration of Dai-ken chu-to and Keish-bukuryo-gan is safe, with no evidence of increased morbidity, and is also highly effective in improving postoperative bowel motility and in reducing hospital stay. PMID- 15783005 TI - Postoperative infections in colorectal cancer patients. AB - BACKGROUND/AIMS: Colorectal surgery is associated with some of the highest rates of infective complications, and especially surgical site infections. It has recently been reported that postoperative infection in colorectal cancer surgery increases the risk of recurrence. The aim of this study was to analyze factors associated with the occurrence of postoperative infections in patients with colorectal cancer. METHODOLOGY: A total of 81 patients operated for colorectal cancer was included. Patients' characteristics and postoperative course were recorded and analyzed. RESULTS: Patients with tumors located in the rectum had significantly higher rate of postoperative infectious complications compared to patients with tumors located in the colon (p=0.002). In a logistic regression model, among all evaluated predictors, only preoperative hemoglobin concentration was found to be an independent significant predictor of postoperative infection (p=0.01). CONCLUSIONS: Preoperative anemia was found to be significant independent predictor of postoperative infection. Meticulous surgical technique with minimal blood loss is an important means of reduction of postoperative infections in colorectal surgery. PMID- 15783006 TI - The pattern of lymphatic metastases in superficial squamous cell carcinoma of the esophagus. AB - BACKGROUND/AIMS: Lymphatic spread patterns in relation to the location of primary tumors of the superficial thoracic esophageal squamous cell carcinoma have not been well established. We therefore analyzed patterns of lymph node metastasis in these patients. METHODOLOGY: We reviewed medical records of 65 patients who underwent systematic three-field dissection for superficial squamous carcinoma of the thoracic esophagus from 1993 through 2000. RESULTS: Lymph node involvement was found in 0% (0/13) and 44% (23/52) of patients whose tumor invaded the muscularis mucosa and submucosal layer, respectively. The 5-year survival rate was 77% in the node-negative group and 59% in the node-positive group (P<0.05). None of the patients with upper thoracic esophageal cancer had metastasis to the mediastinal and abdominal nodes. Patients with lower thoracic esophageal tumors (Lt) had no metastasis to the cervical nodes. Patients with middle thoracic esophageal tumors (Mt) and Lt patients rarely had metastasis (2-5%) in the lower mediasinal nodes (Nos. 108-112). No patient with superficial thoracic esophageal cancer had metastasis to the subcarinal nodes in this study. CONCLUSIONS: In our series, no patient with intramucosal carcinoma had lymphatic metastases. Some patients with submucosal cancers metastasized beyond regional lymph nodes. However, this study suggests that subcarinal nodes might not need to be sampled or dissected in patients with superficial carcinoma of the thoracic esophagus. In Mt and Lt patients, metastases to the mediastinal nodes were infrequent (2-7%). Mediastinal nodes other than #107 can easily be sampled through cervical and abdominal incisions. Therefore, combined with lymph node sampling in cervical, mediasinal and abdominal stations through cervical and abdominal incisions, esophagectomy without thoracotomy might be acceptable in Mt and Lt patients with superficial squamous cell carcinoma of the esophagus. PMID- 15783007 TI - Significance of coupling device for vessel anastomosis in esophageal reconstruction. AB - BACKGROUND/AIMS: To prevent an anastomotic failure due to impaired blood supply, several trials have been performed such as preoperative ischemic conditioning by transarterial embolization of the left gastric, right gastric and splenic arteries or microvascular anastomosis. We assess the significance of an automatic anastomotic coupling device for vessel anastomosis, which we have continuously utilized, to simplify the task and shorten the anastomotic time since March 1999. METHODOLOGY: 8 patients who underwent venous anastomosis by an automatic anastomotic coupling device were evaluated for the time of anastomosis, total ischemic time and outcomes. RESULTS: Venous anastomosis was completed within 5 minutes on average. Microscopic arterial anastomosis by hand took 35 minutes on average. For gastric tube reconstruction, venous anastomosis by an automatic coupling device took only 5 minutes. The top of the gastric tube showed congestion before venous anastomosis, but rapidly recovered from it after anastomosis. Postoperative endoscopic observation of the mucosal color of the replaced intestine or gastric tube was started 3 days after surgery and revealed no ischemia or congestion. The postoperative course was uneventful except one case suffering from pneumonia but leakage was not observed in any case. CONCLUSION: An automatic anastomotic coupling device can perform an easy and reliable vascular anastomosis for patients who undergo esophageal reconstruction. The device may shorten the operating time and consequently the ischemic time of the gastric tube or jejunal or colonic graft, which in turn may lead to a decrease of complications. PMID- 15783008 TI - Intraperitoneal hyperthermic perfusion may induce bacterial translocation. AB - BACKGROUND/AIMS: Intraperitoneal hyperthermic perfusion (IPHP) has been used widely in oncologic practice. Hyperthermia is known to decrease the interstitial pressure. Also intraperitoneal hyperthermia may alter the intestinal mucosal barrier and the intestinal bacterial flora. These changes may lead to bacterial translocation (BT). To the best of our knowledge, there is no data about the possible role of IPHP on BT. METHODOLOGY: Fifty-one rats were divided into two groups. Group I (n=36) received IPHP by heated isotonic salt solution at a temperature of of 43.0 degrees C and group II (n=15) received intraperitoneal normothermic lavage by isotonic salt solution at 37.0 degrees C. Each group was divided into three subgroups which were sacrificed at 1st (Ia, IIa), 3rd (Ib, IIb) and 7th (Ic, IIc) days. Mesenteric lymph nodes (MLN), spleen and liver were sampled and cecal aspirates were obtained. The presence of viable bacteria in samples was noted. Cecal bacterial population levels (CBPL) were reported as colony forming units (CFU). Groups were compared in terms of BT and CBPL. RESULTS: BT was not detected in the Ia (IPHP, 1st day) and IIa,b,c (all control groups). However, statistically significant BT was observed in group Ib and Ic (83.3% and 66.6%, respectively) in comparison to control group (p<0.01). Also there was positive correlation between CBPL and BT. CONCLUSIONS: Intraperitoneal hyperthermia causes remarkable BT. This may explain septic complications after IPHP. Further studies are necessary to better understand the effects of IPHP on the pathophysiology of BT. PMID- 15783009 TI - Non-calculus suppurative cholangitis in Danish patients with inflammatory bowel disease. AB - BACKGROUND/AIMS: We examined the risk of non-calculus suppurative cholangitis in patients with inflammatory bowel disease in the entire Danish population. METHODOLOGY: The study included all patients discharged from Danish hospitals with a diagnosis of Crohn's disease or ulcerative colitis as registered in the Danish National Registry of Patients from January 1, 1977 to December 31, 1992. We compared the observed number of patients hospitalized with suppurative cholangitis with expected numbers on the basis of age, gender, and calendar specific incidence rates in the general population. RESULTS: Overall, 15,317 eligible patients with inflammatory bowel disease were discharged during the study period. Among these were 52 cases of non-calculus suppurative cholangitis. The incidence rate of non-calculus suppurative cholangitis in the cohort with inflammatory bowel disease was 46.1 per 100,000 person-years. The standardized incidence ratio (SIR) for suppurative cholangitis was increased similarly for patients with Crohn's disease [SIR=6.7, 95% confidence interval (CI): 3.1-12.7] and for patients with ulcerative colitis (SIR=6.6, 95% CI: 4.7-9.1). The highest relative risk was found in male patients younger than 40 years of age, for both Crohn's disease and ulcerative colitis (SIR=70.5 and 78.7, respectively). CONCLUSIONS: Patients with inflammatory bowel disease have an increased risk of non-calculus suppurative cholangitis. PMID- 15783010 TI - The role of fibronectin and nitric oxide (NO) in gastrointestinal cancers. AB - BACKGROUND/AIMS: Gastrointestinal cancers are one of the most common malignancies in the world. Various etiologic factors have been proposed including viruses, chemical agents, and genetic factors. The aim of this study was to investigate the role of fibronectin (FN) and nitric oxide (NO) in gastric and colorectal cancers. METHODOLOGY: Thirty-nine patients (22 males, 17 females) with colorectal, 18 (10 males, 8 females) with gastric cancer, and 22 healthy control subjects were included in the study. The Griess reaction was used for the measurement of NO levels. An immunochemical reaction was used for measurement of FN levels. RESULTS: We found increased levels of NO in colon and gastric cancer, and decreased levels of FN in colon cancer when compared with healthy control subjects. Neither FN nor NO levels were associated with age, gender, stage of disease and survival status. No significant association was found between NO and FN levels. CONCLUSIONS: In conclusion, these two molecules might contribute to the pathogenesis of gastrointestinal cancers. The combination with standard chemotherapy and nitric oxide synthase inhibitors may be useful for the treatment of gastrointestinal cancer. PMID- 15783011 TI - Carcinoid tumors of the appendix. Prognostic factors and evaluation of indications for right hemicolectomy. AB - BACKGROUND/AIMS: Carcinoid tumors of the appendix are thought to be the most common type of appendiceal neoplasms. Although the vast majority of appendiceal carcinoids behave in a benign fashion, they are considered malignant because they all have the potential for invasion, metastasis and production of physiologically active substances. The aim of our study is to evaluate the gravity of the prognostic factors and the indications for extended surgical treatment. METHODOLOGY: Twenty-four patients, six male (aged from 18 to 59 years) and eighteen female (aged from 16 to 27 years) are included in our study. All these patients were managed and followed-up in our section during the last 17 years (follow-up range 10-17 years). Following confirmation of the diagnosis, secretion of 5-HLAA (5-Hydroxy-Inndole-Acetic Acid) was measured after a 24-hour collection of urine. Moreover, ultrasound (US) examination of the liver and computerized tomography (CT) scanning of the upper abdomen were performed after the operation in all patients. Also, patients with tumor size more than 1cm underwent a Somatostatin Receptor Scintigraphy (Octreoscan). The follow-up data of all patients included measurement of 5-HLAA and US examination every six months. RESULTS: The most common site for the tumors was the tip of the appendix (18/22). In the rest of the cases, the neoplasms were located in the base (4/22) and in the body (2/22), while in one patient the mesoappendix was invaded. In sixteen patients the tumor size was less than 1cm, in seven patients the tumor diameter was measured to be 1 to 2cm and in one patient the tumor was 3cm. Most of our patients (16/22) underwent only an appendicectomy, while in the rest of them (in the patients with tumor size between 1-2cm and in the patient with invasion of mesoappendix) a right hemicolectomy was performed. No patient was found to have metastatic disease during the operation, while the patient with invasion of the mesoappendix developed metastases in the lung, two years after the operation. CONCLUSIONS: Carcinoid tumors of the appendix, in most cases, are found incidentally during appendicectomies, especially in young females and usually are less than 1cm in size, which is probably the reason of the absence of metastases in all cases. Histological examination and size of the tumor are important factors that contribute to the selection of the surgical treatment and both must be estimated by the surgeons to make the final choice. PMID- 15783012 TI - Endo-laparoscopic approach in the management of obstructive jaundice and malignant gastric outflow obstruction. AB - BACKGROUND/AIMS: Only a minority of patients with tumor at the pancreaticoduodenal junction is suitable for resection, palliation is however often required relieving the obstructive jaundice and gastric outflow obstruction (GOO). This study evaluates endo-laparoscopic approach as a palliative treatment of obstructive jaundice and malignant gastric outflow obstruction. METHODOLOGY: A retrospective review of a prospectively maintained database. During the period from 1992-2002, patients with diagnosis of unresectable tumor at the pancreaticoduodenal junction were evaluated. If the tumor was confirmed to be unresectable, patients would be offered either open double bypass or laparoscopic gastrojejunostomy (LGJ) +/- endoscopic or percutaneous transhepatic stenting for any obstructive jaundice, the choice of approach would depend on whether the endoscopic access was still maintained. RESULTS: Out of 942 patients with tumors around the pancreaticoduodenal junction during the study period from 1992-2002, there were 34 patients (13 male & 21 female) with median age 69 years (range, 48 87) selected for LGJ. Of these 34 patients, 3 of them underwent endoscopic biliary stenting whereas 16 jaundice patients were palliated by transhepatic biliary drainage. When the results were compared to the 35 open double bypass (roux-en-Y choledochojejunostomy and gastrojejunostomy) during the same study period, the median operation time was significantly shorter (80 vs. 135 minutes; P=0.0001) and median intraoperative bleeding was significantly less in the endo laparoscopic group (0 vs. 100mL; P=0.0001). Two patients in the endo-laparoscopic group were converted to open because of tumor infiltration of the small bowel mesentery causing difficulty in construction of gastrojejunostomy. Although the overall complication rate (13 vs. 17; P=0.387) and incidence of delayed gastric emptying (7 vs. 7, P=0.952) were similar in both groups, the incidence of wound infection was remarkably less common in the endo-laparoscopic group (0 vs. 6, P=0.012). The 15 postoperative complications (13 patients) in the endo laparoscopic group (38.2%) included prolonged gastric stasis (7), biliary sepsis (2), chest infection (2), myocardial ischemia (2), gastrointestinal bleeding (1) and extensive ischemic stroke (1). Median time to resume diet was statistically shorter in endo-laparoscopic group (5 vs. 7 days, P=0.009) however the hospital stay was similar in both groups (11.5 vs. 14 days, P=0.238). The hospital mortality rate was again comparable between the two groups (6 vs. 5, P=0.703). The short median survival in the endolaparoscopic group (3 vs. 7 months; P=0.0001) might just be a reflection of selection bias. CONCLUSIONS: With the advent of laparoscopic and endoscopic surgery, palliation of both gastric outflow obstruction and obstructive jaundice can also be accomplished using the endo laparoscopic approach. In comparing to the open double bypass, operation time, intraoperative blood loss and incidence of wound infection are significantly less and patients can have early resumption of diet. However, the results can be improved further with a better patient selection and perioperative optimization. PMID- 15783013 TI - Simple way to improve accuracy in diagnosis of quadrant inflammatory disease: how to avoid adverse laparotomy by using plain CT. AB - BACKGROUND/AIMS: It has been reported that contrast computed tomography (CT) provides a rapid and accurate diagnosis of acute appendicitis, and could potentially prevent adverse appendectomy. In the present study, we evaluated the diagnostic value of precontrast CT combined with that of serum C reactive protein (CRP) concentration for right lower quadrant inflammatory diseases. METHODOLOGY: One hundred consecutive patients who had rebound tenderness and muscular rigidity or guarding on right lower quadrant underwent an emergency abdominal CT without contrast medium and laboratory tests. Based on the CT findings, 10 patients with peritonitis and 41 patients with definite acute appendicitis underwent emergency operations. Of 19 patients with negative CT findings, 11 patients who had positive CRP levels also underwent emergency appendectomies. The remaining patients, of whom 8 had negative CRP levels and 30 had CT findings of other inflammatory diseases, underwent conservative therapy and were examined again later on. RESULTS: The final diagnosis was acute appendicitis in 58 cases, other intestinal diseases in 21, gynecological disease in 6, urolithiasis in 1, and unknown in 14. Among 86 patients who underwent emergency operations and had a pathological diagnosis, CT had a sensitivity of 81% for the diagnosis of appendicitis, with a specificity of 89%, and an accuracy of 84%. CT combined with serum CRP concentration yielded a sensitivity of 100%, an accuracy of 97%, and a specificity of 89%. The adverse appendectomy rate was 3%. None of the patients underwent a delayed appendectomy. CT differentiated other inflammatory diseases from acute appendicitis in 23 patients. CONCLUSIONS: It may be concluded that precontrast CT scan combined with serum CRP can provide a rapid and accurate diagnosis of acute appendicitis and other right lower quadrant inflammatory diseases. PMID- 15783014 TI - Impact of laparoscopy and ultrasonography in gastrointestinal malignancies. AB - BACKGROUND/AIMS: An adequate preoperative disease staging is highly required before surgical treatment, even in gastrointestinal malignancies. Our study wants to give a contribution in order to define echolaparoscopy weight in gastrointestinal tumors and its impact in surgical therapy. METHODOLOGY: 33 patients were affected by pancreas, 22 by stomach, 16 by extrahepatic biliary tract and 18 by liver cancers; every patient was considered worthy of radical or palliative surgery according to preoperative staging (thorax-abdominal CT and percutaneous ultrasonography). Paired sample t-tests were used to analyze the results of each methodical and probability values of less than 0.05 were considered significant. RESULTS: Preoperative instrumental examinations gave correct evaluations only in 44 of 89 cases (49%) while echolaparoscopic gave correct evaluations in 82 on 89 cases (92%) (P<0.05). So after echolaparoscopic in only 7 cases we performed an explorative laparotomy. CONCLUSIONS: Laparoscopy and ultrasound impact on therapy is worthy of attention. It seems to be able to give advantages in staging gastrointestinal malignancies, except for pancreas cancers, in which some limits and negative aspects have been demonstrated, regarding the possibility of giving correct diagnosis of portal axis infiltration. PMID- 15783015 TI - Clinical and surgical factors influencing delayed gastric emptying after pyloric preserving pancreaticoduodenectomy. AB - BACKGROUND/AIMS: Delayed gastric emptying (DGE) is the most common and troublesome complication after pylorus-preserving pancreaticoduodenectomy (PPPD), however, definitive treatment has not yet been established. We examined the clinical and surgical factors relevant to DGE using multivariate analyses. METHODOLOGY: Forty-four patients with PPPD were divided into two groups according to reconstructive technique: group A (25), Billroth II type with antecolic duodenojejunostomy and group B (19), Billroth-I type. Multiple clinical and surgical factors influencing DGE were evaluated by univariate and multivariate analyses. RESULTS: The period and output of gastric aspiration were significantly reduced in group A compared with group B (a median of 3 days vs. 14 days and a mean output of 133+26mL vs. 506+80mL, respectively; p<0.0001). Re-insertion of the tube was required in 8% of group A compared with 32% of group B. A liquid or solid diet was started at medians of 8 and 14 days in group A compared with 22 and 28 days in group B (p<0.0001), respectively. Multivariate analyses disclosed that the antecolic duodenojejunostomy and major complication were two exclusive independent predictors of restoration of gastric motility. CONCLUSIONS: Occurrence of DGE was strongly affected by reconstruction technique and major complication. Billroth II reconstruction with antecolic duodenojejunostomy seems to be a useful technique to minimize the occurrence of DGE. PMID- 15783016 TI - Peripheral portosystemic shunt and its selectivity changes measured on duplex ultrasound. AB - BACKGROUND/AIMS: Portosystemic shunts offer a symptomatic treatment for portal hypertension. Their main disadvantage is decreased perfusion of the liver with portal blood. Change of peripheral shunts into total shunts after a period of time is described. This study aims to evaluate long-term hemodynamic changes in peripheral portosystemic shunts. METHODOLOGY: The study was based on 12 patients in whom distal splenorenal shunts 8 patients) and mesocaval shunts (4 patients) were indicated respectively. Duplex sonography was used to measure the blood flow in the portal, splenic and mesenteric veins before shunt surgery and minimally 14 months postoperatively. RESULTS: It was found that the reduction of the portal blood flow was not critical and no centralization of the shunt was observed. CONCLUSIONS: Long-term blood flow in the portal vein was not severely reduced after peripheral portosystemic shunt creation, therefore the peripheral portosystemic shunt still has a role in the treatment of some patients with portal hypertension. PMID- 15783017 TI - Appraisal of DSRS with SPGD for esophagogastric varices: a retrospective comparative study according to the underlying liver diseases. AB - BACKGROUND/AIMS: The differences in long-term results of distal splenorenal shunt with splenopancreatic and gastric disconnection (DSRS with SPGD) for portal hypertension of different etiologies including non-cirrhotic portal hypertension have yet to be reported. The data are important to determine the indications and contraindications for this procedure. METHODOLOGY: Records of 54 patients of esophagogastric varices who survived 3 years or longer after DSRS with SPGD operation were reviewed. Patients were divided into three groups based on underlying liver disease; posthepatitic cirrhosis (HC) group, alcoholic cirrhosis (AC) group, and idiopathic portal hypertension (IPH) group. RESULTS: The only serious long-term complication of DSRS with SPGD was portal thrombosis in two patients in the IPH group. Postoperative bleeding occurred in two cases of each group; one in IPH group was the only variceal bleeding and others were bleeding from portal hypertensive gastropathy. Hepatocellular carcinoma (HCC) was developed in 28.6% patients in both the HC group and AC group. In all the cases, treatment for HCC was accomplished without aggravation of the varices. The cumulative survival rate was similar in the three groups, and no patient died of gastrointestinal bleeding. CONCLUSIONS: A favorable outcome was achieved by DSRS with SPGD operation both in the patients with cirrhosis or IPH. Underlying liver disease is not a factor when considering DSRS with SPGD for portal hypertension. PMID- 15783018 TI - Doppler hemodynamic study in portal hypertension and hepatic encephalopathy. AB - BACKGROUND/AIMS: The aim of our study was to evaluate and compare the differences in the parameters of portal hypertension in two groups of patients with liver cirrhosis, with and without hepatic encephalopathy (HE). METHODOLOGY: 30 patients with liver cirrhosis, 17 (56.7%) of them with HE, were investigated by clinical, neurological, laboratory, endoscopic methods and with color Doppler ultrasonography (CDU) at the Institute for Digestive Diseases, Clinical Center of Serbia, Beograde. RESULTS: Significant correlation was found between the diameters of the right liver lobe and the portal vein (p=0.01), and also between the diameters of the spleen and splenic vein (p=0.0002), in both groups of patients. Mean portal vein diameter significantly increases (p=0.01) in patients with HE (14.87 +/- 1.86mm), compared to those without HE (13.2 +/- 2.31mm), while mean splenic vein diameter was not significantly different in the two groups. In patients with ascites, CDU showed significantly lower (p=0.03) portal flow velocity (11.87 +/- 6.25cm/ sec), compared to those without ascites (14.33 +/- 4.41cm/sec). Splenic flow velocity was not significantly different (16.00 +/- 6.60cm/sec with ascites and 14.61 +/- 5.29cm/sec without ascites). In patients with HE, portal flow velocity was significantly lower (9.00 +/- 5.41cm/sec) compared to those without HE (14.0 +/- 7.03cm/sec) (p=0.04). Mean splenic flow velocity was significantly lower (p=0.03) in patients with HE (12.60 +/- 4.16cm/sec), compared to those without HE (17.77 +/- 5.91cm/sec). Portal flow velocity shows linear decrease, related to the increase of the liver damage (Child-Pugh score), while splenic velocity was not related to this parameter. CONCLUSIONS: Ultrasonographic parameters of portal hypertension show significant correlation between the diameters of liver/portal vein and spleen/splenic vein. Portal hemodynamic parameter (blood flow velocity) is significantly related to the stages of liver damage, presence of ascites and HE, while splenic hemodynamics is specific and not directly related to these parameters. PMID- 15783019 TI - Effect of the herbal medicine Dai-kenchu-to for serum ammonia in hepatectomized patients. AB - BACKGROUND/AIMS: Prolonged paralytic ileus occurring in hepatectomized patients may induce hyperammonemia or bacterial translocation, which injures the remnant liver function and sometimes causes post-resection liver failure. We examined the effectiveness of the herbal medicine, Dai-kenchu-to (DKT), on postoperative serum ammonia levels in patients with liver resection and compared it with lactulose. METHODOLOGY: Patients with liver resection were divided into three groups. Lactulose group (n=31), 16g of lactulose was administered orally three times a day from the first postoperative day. DKT group (n=27), 5g of DKT was administered in the same fashion. Control group (n=26), neither lactulose nor DKT was administered. In all three groups, 16g of lactulose was administered three times a day for three days preoperatively. RESULTS: There was no significant difference among the groups in age, gender and preoperative hepatic functional values, such as ICG-R15 or galactose tolerance test. There was also no difference in parenchymal hepatic resection rate, operative time and amount of intraoperative bleeding volume. Postoperative serum ammonia levels were significantly lower in the DKT group than control and lactulose groups. Instances of delayed flatulence and occurrence of diarrhea were also fewer in the DKT group. CONCLUSIONS: DKT may become a more effective and safe agent than lactulose in postoperative management of liver resection. PMID- 15783020 TI - The clinical experience of Chinese patients with Wilson's disease. AB - BACKGROUND/AIMS: Wilson's disease has protean presentations. Most patients manifested as either hepatic or neurologic symptoms and signs initially. No study has investigated the predictor of presentation and differences in laboratory tests and survival between patients presented with hepatic or neurological disease so far. METHODOLOGY: Forty-one patients with Wilson's disease were diagnosed between 1981 and 2002 at our hospital. Their clinical features at presentation and laboratory findings were retrospectively collected. Survival was calculated using the Kaplan-Meier analysis. RESULTS: Twelve patients initially presented with hepatic symptoms including chronic hepatitis, complications of cirrhosis, and fulminant hepatic failure. Twenty-nine patients presented with neurological symptoms including tremor, dysarthria, ataxia, and psychiatric diseases. No specific factor identified could predict the initial presentations. Compared to patients with neurological symptoms, patients with hepatic disease had significantly higher serum levels of aminotransferase, ceruloplasmin, copper, and 24-hour urine copper excretion. There was no significant difference in survival between these two groups. Among the patients who died, the most common causes of death were fulminant hepatic failure and complications of cirrhosis. CONCLUSIONS: Our results suggested that the overall survival did not differ significantly according to the initial presentations. There was no predictor found that could foretell the clinical course of Wilson's disease. PMID- 15783021 TI - Safe bile duct division in right lateral sector graft. AB - BACKGROUND/AIMS: Right lateral sector (RLS, segments VI and VII according to Couinaud's nomenclature for liver segmentation) graft was introduced as an alternative liver graft. The authors report their surgical techniques for bile duct division in RLS graft harvest. METHODOLOGY: Between January 2000 and December 2001, 11 donors provided their RLS. Intraoperative cholangiography was performed and the anatomy of the upper biliary confluence was confirmed. When the right lateral duct entered the common bile duct (caudal right lateral duct), the duct was divided at its root in situ. In other situations, the right portal branch was first dissected and taped. With the right portal branch pulled cranially, the right lateral duct was dissected from surrounding connective tissues and divided as long as possible. RESULTS: Cholangiography revealed various right lateral duct anatomies, such as entering the right duct (n=6), the left and right bifurcation (n=2), the common bile duct (n=2) and the left duct (n=1). The postoperative course was uneventful in all donors. All of the patients survived the operation. CONCLUSIONS: Careful attention should be paid to bile duct division in donor hepatectomy. The present technique can ensure a safe harvest of RLS graft with a long right lateral duct. PMID- 15783022 TI - Role of ultrasonically activated scalpel in hepatic resection: a comparison with conventional blunt dissection. AB - BACKGROUND/AIMS: Our goal was to compare the benefits and complications of using an ultrasonically activated scalpel (UAS) and conventional blunt dissection in hepatic resection. METHODOLOGY: We evaluated the effectiveness of dividing the liver by UAS (n=18) (the UAS group) compared with conventional blunt dissection (n=34) (the BD group) in patients undergoing hepatic resection. In the UAS group, UAS was used to dissect the superficial parenchyma and a crushing and clamping technique was used to divide the deep parenchyma. RESULTS: No serious complications attributable to the use of UAS were encountered, and there were no significant differences in morbidity or mortality between the two groups. The duration of surgery was significantly longer in the UAS group (281 +/- 81 min) than in the BD group (223 +/- 76 min) (P<0.05), and in the UAS group as a whole there were no advantages in using the new scalpel. However, when we compared only those patients who underwent minor hepatectomy, the intraoperative blood loss was significantly less in the UAS group (657 +/- 588mL) than in the BD group (1447 +/ 984mL) (P=0.03). The duration of drainage from the hepatic stump in these patients was also significantly shorter in the UAS group (P=0.02). CONCLUSIONS: The UAS is a useful new device for transection of the liver during hepatic resection. It may reduce the amount of blood loss during liver surgery, particularly in minor hepatectomy. PMID- 15783023 TI - Factors associated with the development of cirrhosis in patients with HCV chronic infection. AB - BACKGROUND/AIMS: The natural history of chronic hepatitis C virus (HCV) infection still has some details to be established, namely in what concerns progression to hepatic cirrhosis (HC). The study aims to define predictive factors for progression to HC in patients with HCV chronic infection. METHODOLOGY: A cross sectional study was performed on 129 patients consecutively submitted to liver biopsy. Thirty-six percent (n=46) had HC at histological evaluation. RESULTS: Patients with HC did not show statistically significant differences on gender, viruses genotypes, alcohol consumption or proportion of positivity to markers of previous hepatitis B virus (HBV) infection - anti-HBc/anti-HBs+. Patients with HC seem to have had their infection sporadically (50%) or post-transfusion (35%) p=0.052, and iv drugs addiction was related to non-HC patients (39%) -p=0.006. Age at infection, time of infection and positivity for anti-HBc/anti-HBs were factors independently related to HC (multivariate analysis). Patients older than 40 years by the time of infection [OR=4.5 (95% CI=1.9-10.8], those with less than 5 years of time of infection [OR=4.2 (95% CI=1.6-10.8)], and patients with previous HBV infection [OR=2.51 (1.00-6.69)] are at higher risk for HC. CONCLUSIONS: We argue that older patients, with a shorter time interval between HCV infection and diagnosis, and namely those with markers for previous HBV infection represent patients with higher risk for progression to hepatic cirrhosis. PMID- 15783024 TI - Prevalence of obesity in patients with acute hepatitis; is severe obesity a risk factor for fulminant hepatitis in Japan? AB - BACKGROUND/AIMS: The prevalence of obesity in acute convalescent hepatitis and fulminant hepatitis has not been reported. The aim of this study was to investigate whether obesity affected the disease severity in Japanese patients with acute hepatitis. METHODOLOGY: 31 non-severe acute hepatitis (NS-AH) and 24 severe acute hepatitis and 14 fulminant hepatitis patients (S-AH) between January 1995 and December 2001 were analyzed retrospectively. Height and weight were used to calculate the body mass index (BMI) in these 69 patients. RESULTS: Mean height, weight and BMI were not significantly different between S-AH and NS-AH patients. Two severely obese (BMI greater than 35kg/m2) patients had developed S AH. CONCLUSIONS: Severe obesity may be one of the prognostic factors in acute hepatitis. Further studies are needed. PMID- 15783025 TI - Surgical approach to symptomatic giant cavernous hemangioma of the liver. AB - BACKGROUND/AIMS: Surgical treatment of giant cavernous hemangioma of the liver is still controversial. In this study, indications and results of surgical therapy were evaluated. METHODOLOGY: Fifteen patients with symptomatic giant cavernous hemangioma of the liver were treated by enucleation or liver resection. RESULTS: The surgical indications were abdominal pain in 11 patients, uncertain diagnosis in 3 patients and tumor enlargement in one patient. The median tumor size was 12.5cm (range, 6-30cm). Eleven patients underwent enucleation procedure while the other 4 patients underwent resection procedures. Complications occurred in 2 (13.4%) patients. The patient with the largest tumor underwent right extended lobectomy and died of bleeding and coagulopathy (6.7%). The postoperative hospital stay was 7 days (range, 4-16 days). Thirteen patients were followed-up for an average period of 32.8 months (range, 6-88 months). It was found that the symptoms for 12 of 13 patients disappeared. During the postoperative controls carried out by imaging procedures, no recurrences were observed. CONCLUSIONS: Abdominal pain, uncertain diagnosis and enlargement are major surgical indications of symptomatic giant cavernous hemangiomas. Most of the symptoms disappear after the surgical treatment. Enucleation can be successively performed with low morbidity rates in most of the patients and recurrences are rare. If the tumor location precludes safe enucleation, anatomic resections are preferred. PMID- 15783026 TI - Role of tissue trace elements in liver cancers and non-cancerous liver parenchyma. AB - BACKGROUND/AIMS: The role of trace elements in liver fibrosis, carcinogenesis and progression of hepatocellular carcinoma (HCC) has not yet been clarified. The aim of this study is to analyze the characteristics of trace elements in liver cancers and non-cancerous liver and to discuss their role in hepatic fibrosis, hepatocarcinogenesis and progression of HCC. METHODOLOGY: The amount of zinc (Zn), iron (Fe), and copper (Cu) in 20 HCCs, 2 cholangiocellular carcinomas (CCC), 7 metastatic liver cancers (Meta) and their non-tumorous liver parenchyma were measured using an atomic absorption spectrophotometer. RESULTS: The amounts of Zn and Fe in non-tumorous liver parenchyma were reduced by liver fibrosis, and the amounts were lower in HCC tissue compared to non-tumorous liver parenchyma. The amounts of Zn and Cu were higher in HCC than the amounts found in CCC and Meta. The amount of Zn in HCC tissue decreased, but the amount of Fe increased in tumors more than 4cm in diameter. CONCLUSIONS: These results suggest that the decrease in the amount of Zn and Fe found in non-tumorous liver parenchyma correlates with liver fibrosis leading to cirrhosis and hepatocarcinogenesis. Also that decreases in Zn and increases of Fe in HCC tissue correlates with HCC tumor progression. PMID- 15783027 TI - Clinical significance of portal venous metallic stent placement in recurrent periampullary cancers. AB - BACKGROUND/AIMS: We evaluate the clinical significance of portal venous expandable metallic stent (EMS) placement for patients who have malignant portal hypertension caused by recurrent periampullary cancer. METHODOLOGY: Four post pancreatoduodenectomy patients underwent percutaneous transhepatic portal venous EMS placement because of symptoms of malignant portal hypertension: intractable ascites (2/4), growth of abnormal collateral vein (3/4), melena (2/4), gastroesophageal varix (3/4) and thrombocytopenia (2/4). They were diagnosed with having a recurrence by cytology of ascitis, computed tomography and/or tumor markers in serum. The stenosis segment was measured by percutaneous-transhepatic portography and was dilated with a balloon prestent placement. The patency of stent was confirmed using Doppler ultrasonography and enhanced computed tomography at least once a month. RESULTS: The portal venous pressure was significantly decreased from (24.5 +/- 3.92 mmH2O) to (19.5 +/- 3.87 mmH2O) and the symptoms related to portal hypertension were reduced in all patients. There were no complications related to EMS placement. All patients were alive more than a year later and two of four patients were alive more than two years without morbidity. CONCLUSIONS: Percutaneous-transhepatic portal EMS placement is a minimally invasive procedure and is a useful treatment against malignant portal hypertension caused by recurrent periampullary cancer. PMID- 15783028 TI - Implantable Doppler probe for continuous monitoring of blood flow after liver transplantation. AB - BACKGROUND/AIMS: Hepatic artery and portal vein thromboses after liver transplantation remain significant causes of graft loss. Doppler ultrasonography is useful for detecting the thrombosis early. Doppler examination requires specialized equipment and training. However the results depend on the skill of the operator. METHODOLOGY: We developed a new Doppler probe that can be implanted in the abdominal cavity. The probe was used for 48 hours in 10 living donor liver transplantation patients. RESULTS: In all recipients, hepatic arterial and portal flows were monitored continuously. The medical staff could easily operate the ultrasonography system at the bedside. CONCLUSIONS: The technique was useful for detecting hepatic blood flow for 48 hours after liver transplantation. PMID- 15783029 TI - Risk factors of cytomegalovirus infection after living donor liver transplantation. AB - BACKGROUND/AIMS: Cytomegalovirus (CMV) infection is one of the most common infectious diseases complicating liver transplantation. Data regarding the incidence and results after CMV infection or disease after living donor liver transplantation, however, are limited. METHODOLOGY: A total of 169 patients who underwent living donor liver transplantation in. our department were enrolled in the study. All of the patients were preemptively treated with intravenous ganciclovir (10mg/kg/d) when the pp65 antigenemia assay was positive. The incidence of CMV infection and disease, and risk factors for CMV infection were evaluated. RESULTS: The incidence of CMV infection and CMV disease was 44% and 3%, respectively. Acute rejection was significantly associated with CMV infection. CONCLUSIONS: The incidence of CMV disease was low, suggesting that our strategy might be effective for preventing the development of CMV disease. PMID- 15783030 TI - Hepatobiliary scintigraphy after biliary reconstruction-Roux Y and RY-DJ. AB - BACKGROUND/AIMS: Reconstruction of extrahepatic biliary tract with benign lesion still has some unsettled problems, such as postoperative cholangitis. This study was conducted to compare bile through the remnant alimentary tract in patients undergoing Roux-Y-duodenojejunal anastomosis (RY-DJ) which was designed for decompressing the jejunal limb of R-Y and for allowing an inflow of bile into the duodenum, and those undergoing Roux-Y choledochojejunostomy (R-Y), using hepatobiliary scintigraphy. METHODOLOGY: Five normal human volunteers and 20 patients underwent R-Y (n=14), RY-DJ (n=6), using hepatobiliary scintigraphy. RESULTS: Postoperative cholangitis developed in 2 patients (14%) with R-Y and none with RY-DJ. Hepatobiliary scintigraphy showed prominent stasis of 99mTc in the proximal jejunum loop of the patients who underwent R-Y, which was not found in the upper jejunum of the patients with RY-DJ. The time taken before visualization of 99mTc at the upper jejunum in the patient who underwent R-Y (65 +/- 5 min) was significantly longer than that in the healthy control (40 +/- 5 min). On the other hand, the time taken before visualization of 99mTc at the upper jejunum in RY-DJ (45 +/- 5 min) was similar to that of healthy controls. CONCLUSIONS: These data suggested that this new method (RY-DJ) for reconstructing the extrahepatic biliary tract was more physiological with less postoperative complications than R-Y. PMID- 15783032 TI - Middle hepatic vein tributary reconstruction could not act as a complete substitute for an entirely preserved middle hepatic vein. AB - BACKGROUND/AIMS: The necessity of the middle hepatic vein for living donor liver transplantation using right lobe graft is still controversial. METHODOLOGY: We reviewed 7 long-term surviving right-lobe recipients in whom middle hepatic vein tributaries were not reconstructed (group A, n=4) or were reconstructed (group B, n=3). Volume regeneration of the right paramedian (segments V+VIII) and right lateral (segments VI+VII) sectors was assessed by computed tomography at 3, 6, 9, and 12 postoperative months. The right paramedian sector was further subdivided into the ventral portion in relation to the anterior branch of the right portal vein and dorsal portion. RESULTS: The volume regeneration ratio was significantly lower in group A than in group B persistently after 6 postoperative months in regard to the right paramedian sector, the dorsal portion, and especially the ventral portion (0.64 +/- 0.19 vs. 1.22 +/- 0.17, p=0.034, 12 postoperative months). However, volume regeneration was impaired in the ventral portion as compared to other areas in group B. CONCLUSIONS: In conclusion, middle hepatic vein tributary reconstruction improves the volume regeneration of the right paramedian sector in right lobe living donor liver transplantation However, it could not act as a complete substitute for an entirely preserved middle hepatic vein. PMID- 15783031 TI - Quantitative evaluation of magnetic resonance imaging (MRI) abnormalities in subclinical hepatic encephalopathy. AB - BACKGROUND/AIMS: Cirrhotic patients often demonstrate high signal intensity on T1 weighted magnetic resonance (MRI) images in basal ganglia with accumulation of manganese being the predominant causing factor. In these patients, electrophysiological tests and especially electroencephalogram (EEG) are considered to be the most sensitive methods in detection of subclinical hepatic encephalopathy. The aim of this study is to correlate MRI findings with biochemical parameters and EEG alterations in cirrhotic patients without clinically overt encephalopathy. METHODOLOGY: Twenty-two cirrhotic patients (16 males and 6 females, mean age of 65.2 +/- 9.5 years), classified according to Child-Pugh score, were submitted to brain MRI, neurological assessment (including psychometric tests and EEG) and complete biochemical testing. None of them had any clinical signs of brain dysfunction. MRI findings were evaluated both qualitatively (normal, mild, moderate and severe) and quantitatively with the ROI method. EEG alterations were also classified as normal, mild, moderate and severe. RESULTS: Statistical analysis revealed a significant linear association between EEG grading and MRI signal intensity (r2=0.248, p=0.035). Among clinical and biochemical parameters, overall Child-Pugh score and albumin levels were identified as significant predictors of the MRI signal intensity (p=0.006 and p=0.021 respectively). CONCLUSIONS: Although further investigation must be performed to confirm the clinical impact of brain MRI in hepatic cirrhosis, our study strongly suggests that MRI alterations are good predictors of liver and brain dysfunction in cirrhotic patients. PMID- 15783033 TI - Does a surgery for hepatocellular carcinoma with tumor thrombus highly occupying in the right atrium have significance? A case report and review of the literature. AB - Hepatocellular carcinoma (HCC) advancing to the right atrium (RA) through the hepatic vein has generally been regarded as a terminal lesion of carcinoma. However, because tumor thrombus (TT) in the RA may cause sudden death by heart failure or pulmonary embolism, it is a pathologic condition that needs emergency extraction. Our case was a 55-year-old male. The HCC had a main lesion in the anterior region of the hepatic right lobe, accompanying TT highly occupying the region from the middle hepatic vein and inferior vena cava to the RA. For this tumor, we conducted an extended right anterior segmentectomy and extraction of the TT in the RA under an extracorporeal circulation. He was discharged on the 28th day after surgery, and at present, when 12 months have passed since the surgery, survives without any sign of its recurrence. There have been eight HCC cases including our case, reported regarding the simultaneous resections of a main tumor and TT under cardiopulmonary bypass. Because two patients among this group of eight survived for more than two years, resection is recommended even for advanced HCC highly infiltrating to the RA. PMID- 15783034 TI - Prolonged survival after liver transplantation for Hodgkin's disease-induced fulminant liver failure. AB - A 30-year-old male with a past history of nodular lymphocyte predominance Hodgkin's disease in apparent complete remission for two years received a liver transplantation because of fulminant liver failure. Histopathological examination of the explanted liver showed massive infiltration by Hodgkin's disease. In spite of a nodal recurrence of Hodgkin's disease, the patient is alive and in excellent general condition six years after liver transplantation. PMID- 15783035 TI - Repeated obstructive jaundice and acute pancreatitis caused by metastatic liver tumor: an unusual case. AB - A case of repeated obstructive jaundice and acute pancreatitis caused by mucus plug produced by metastatic liver tumor is reported. A 74-year-old woman, who had a past history of curative resection of mucinous rectal cancer, showed repeated obstructive jaundice and acute pancreatitis during the follow-up period. Neither computed tomographic scan nor abdominal ultrasound could detect the recurrent lesion, however, cholangioscopy detected mucin and tumor projection into the left hepatic duct. Since the biopsy specimen of the tumor revealed adenocarcinoma, left hepatectomy was performed. The tumor was mucinous adenocarcinoma having the same histology as the primary rectal cancer, with partly mucosal replacement and formation of intraluminal mucus plaque. This case indicates that repeated obstructive jaundice and acute pancreatitis should be considered one of the manifestations of liver metastasis of mucinous cancer. PMID- 15783036 TI - Pancreatitis associated with congenital abnormalities of the pancreaticobiliary system. AB - BACKGROUND/AIMS: The relationship between pancreas divisum and pancreatitis remains controversial. The aim of this study was to examine the frequency and features of pancreatitis associated with congenital abnormalities of the pancreaticobiliary system, and the role that these malformations have in the occurrence of pancreatitis. METHODOLOGY: We retrospectively examined endoscopic retrograde cholangiopancreatograms of 6850 patients including 186 patients who were diagnosed clinically as having acute pancreatitis and 211 who were diagnosed as having chronic pancreatitis. RESULTS: A total of 175 patients were diagnosed as having congenital abnormalities of pancreaticobiliary system. Acute and chronic pancreatitis was associated respectively with 3.0% and 5.0% of patients with pancreaticobiliary maljunction (n=100), 17.1% and 28.6% of those with pancreas divisum (n=35), 6.6% and 20.0% of those with partial pancreas divisum (n=30), and 16.7% and 33.3% of those with choledochocele (n=6). These anomalies were present in 6.5% and 10.9% of patients with acute and chronic pancreatitis, respectively. Isolated chronic dorsal pancreatitis was detected in 8 patients with pancreas divisum. In chronic pancreatitis associated with pancreaticobiliary maljunction, pancreatic stones or protein plugs were situated only in the dilated common channel or main pancreatic duct of the head. CONCLUSIONS: Congenital abnormalities of the pancreaticobiliary system constitute one etiology of pancreatitis. PMID- 15783037 TI - Laparoscopic pancreatic resection: a preliminary experience of 15 patients. AB - BACKGROUND/AIMS: Worldwide experience with laparoscopic pancreatic resection remains limited. The aim of the study was to assess the feasibility, safety and outcome of laparoscopic pancreatic resection. METHODOLOGY: 15 consecutive patients suffering from benign cystic pancreatic (n=6), neuroendocrine tumors (n=8) or pancreatic metastasis from renal carcinoma (n=1) undergoing laparoscopic pancreatic resection were retrospectively collected from 5 academic hospitals. RESULTS: Laparoscopic procedure was completed in 10 patients, including 7 distal pancreatectomies (with 5 spleen preservation), 2 tumor enucleations and 1 partial cystic resection. Conversion was due to inappropriate operative finding for laparoscopic approach in 2 patients and for uncontrollable bleeding in 3 patients. Postoperative pancreatic-related complications included pancreatic fistula in 20% and peripancreatic collection in 13% of the patients. CONCLUSIONS: Laparoscopic pancreatic resection is feasible for distal pancreatic tumors. However, successful management of the pancreatic stump remains the challenge of this procedure, in order to achieve a clear benefit in the patient outcome. PMID- 15783038 TI - Postoperative pancreatic fistula following surgery for gastric and pancreatic neoplasm; is distal pancreaticosplenectomy truly safe? AB - BACKGROUND/AIMS: A pancreatic fistula is a potentially fatal complication in digestive tract surgery. The aim of this study was to review the prevention and treatment of pancreatic fistula as performed by our department. METHODOLOGY: Three hundred and seventeen patients with gastric neoplasm and 34 patients with pancreatic neoplasm who underwent surgical resection between 1981 and 2001 at Kochi Medical School were studied. Comparative analysis of the incidence of pancreatic fistulae and the mortality rate was carried out according to an operation type and time. RESULTS: Among the 317 patients who underwent total gastrectomy due to gastric neoplasm, 30 (9.5%) showed pancreatic fistulae. The incidence of pancreatic fistulae in patients who underwent distal pancreatectomy for pancreatic neoplasm was 29.4%. The mortality rate of patients who underwent total gastrectomy was 4.4% and there was a tendency for mortality rate to decrease in the second decade compared to the first. The mortality rate of patients who underwent distal pancreatectomy was 5.9%. CONCLUSIONS: Mortality rate tended to decrease with advanced operative procedures, however, the incidence of pancreatic fistulae did not decrease. In pancreatic fistulae, it was found that the type of treatment was important. Intraoperative and postoperative drainage management for pancreaticosplenectomy will likely become much more important. PMID- 15783039 TI - Intraportal endovascular ultrasound for portal vein resection in pancreatic carcinoma. AB - BACKGROUND/AIMS: Intraportal endovascular ultrasound (IPEUS) has been reported to be the most precise diagnostic procedure for the accurate diagnosis of portal vein/superior mesenteric vein (PV/SMV) invasion in patients with pancreatic cancer. In this study, we evaluated the clinical significance of the length of PV/SMV invasion measured by IPEUS METHODOLOGY: Twenty-six consecutive patients who underwent the pancreatic resection and IPEUS using an autopull-back device between January, 1997 and September, 2000 were retrospectively evaluated. The length of PV/SMV invasion was measured by reviewing the videotapes recorded during the operation. Clinicopathological data and survival were analyzed. RESULTS: The percentage of PV/SMV invasion was 46%, all of which were treated by PV/SMV resection. The cases without PV/SMV invasion showed significantly longer survival rate. The cases with < or = 18mm PV/SMV invasion, however, achieved a comparable 2-year survival rate of 28% whereas no patient with > 18mm PV/SMV invasion survived more than 18 months after the resection. CONCLUSIONS: Involvement of the PV/SMV by pancreatic carcinoma seems to be related to the extent of the disease and the PV/SMV involvement > 18mm is associated with a poor prognosis due to high rate of tumor positive margin even with radical operation. PMID- 15783040 TI - Value of procalcitonin quick test in the differentiation between sterile and infected forms of acute pancreatitis. AB - BACKGROUND/AIMS: Authors analyzed the clinical value of procalcitonin quick test (PCT-Q; BRAHMS Diagnostica, GmbH, Berlin) in infected pancreas necrosis verified by guided fine-needle aspiration (FNA). METHODOLOGY: In the prospective, controlled study data of 24 patients were evaluated. PCT-Q was performed in patients with necrosis--verified on computer tomography (CT scan)--and/or sepsis. If PCT-Q test was positive or septic complication (infected necrosis or abscess) developed, CT or ultrasound (US) guided fine-needle aspiration was performed with Gram staining and bacteriology. Positive FNA result was indication for surgery with repeated staining and bacteriology of the surgical specimen. RESULTS: Septic complications developed in 12 patients. Analyzing the results, fine-needle aspiration was more authentic with 92% sensitivity and 100% specificity, while sensitivity of PCT-Q test remained 75% and its specificity 83%. Comparing abscess with infected necrosis significantly higher procalcitonin values were detected in patients with necrosis. CONCLUSIONS: These results show that PCT-Q test is a possible non-invasive method which can be used besides fine-needle aspiration. Elevated levels of procalcitonin (higher than 2ng/mL) clearly suggest infection, while lower values do not exclude the possibility of local sepsis. PMID- 15783041 TI - Dose escalation of concurrent hypofractionated radiotherapy and continuous infusion 5-FU-chemotherapy in advanced adenocarcinoma of the pancreas. AB - BACKGROUND/AIMS: To investigate the advantages and palliative effectiveness of concurrent hypofractionated radiotherapy (RT) and chemotherapy (5-FU) in patients with locally advanced and metastatic adenocarcinoma of the pancreas. METHODOLOGY: A total of 26 patients were enrolled in this study. Twenty patients had locally advanced (M0) and 6 patients had metastatic (M1) disease. They were treated with hypofractionated radiation therapy (RT) (4x3 Gy per week) and concurrent continuous infusion (300mg/sqm/24h) of 5-fluorouracil. The RT doses were escalated in 6-Gy increments starting from 24 Gy in 8 fractions in 2 weeks to 30 Gy in 10 fractions in 2.5 weeks and finally to 36 Gy in 12 fractions in 3 weeks. RESULTS: Only 1 (4%) patient experienced grade 3 mucositis, while 12 (46%) patients experienced grade 2 nausea and 1 (4%) patient experienced grade 2 weakness. No patient experienced treatment interruption or dose reduction. Late high-grade (>3) toxicity was not observed, but few patients experienced prolonged hematological toxicity, due to administration of chemotherapy after radiochemotherapy. Pain improved in 70% of the patients. The median survival time for all 26 patients is 8 months, 9 months for locally advanced cancer patients and 5 months for metastatic cancer patients. CONCLUSIONS: Dose escalation to 36 Gy in a hypofractionated manner proved to be feasible with low toxicity in patients with locally advanced and metastatic adenocarcinoma of the pancreas and warrants further investigation aiming at optimal tailoring in these two subgroups of patients. PMID- 15783042 TI - How necessary is vagotomy after pancreaticoduodenectomy and total pancreatectomy. AB - BACKGROUND/AIMS: Gastrectomy with gastrojejunostomy is a fundamental step used for the restoration of the alimentary tract after pancreaticoduodenectomy or total pancreatectomy. Anastomotic ulcers occurring after pancreaticoduodenectomy, is a well known problem. The aim of our study is to investigate the incidence of anastomotic ulcer after pancreaticoduodenectomy or total pancreatectomy and to elucidate whether vagotomy is necessary. METHODOLOGY: In this study we reviewed the medical records of 94 patients who underwent pancreaticoduodenectomy or total pancreatectomy without vagotomy and we report the results after systemic follow up 3-14 years, emphasizing the cases in which anastomotic ulcer is jeopardized. RESULTS: A total of 78 Whipple procedures and 16 total pancreatectomies without vagotomy were performed. The overall incidence of anastomotic ulceration following pancreatectomy was 11.7%. In our series the symptoms and complications associated with anastomotic ulceration were pain in 6 patients, bleeding in 4 patients and free perforation in one patient. Six patients needed a reoperation, the 4 patients with bleeding that underwent truncal vagotomy, the patient with free perforation in which oversew and bilateral vagotomy was performed and a patient with refractory pain who underwent a bilateral vagotomy. Among the 11 patients with anastomotic ulcer, the overall postoperative mortality rate was 27.3%. CONCLUSIONS: It seems reasonable to perform bilateral truncal vagotomy only in patients with a history of peptic ulceration and for patients with favorable prognosis and potential for long survival. PMID- 15783043 TI - Non-stented pancreaticogastrostomy for 111 patients undergoing pylorus-preserving pancreaticoduodenectomy. AB - BACKGROUND/AIMS: The main purpose is to clarify the roles of pancreatic stenting and duct-to-mucosa anastomosis in prevention of pancreatic leakage and exocrine insufficiency in pylorus-preserving pancreaticoduodenectomy with non-stented pancreaticogastrostomy (non-stented PPPD-PG). METHODOLOGY: Prospectively collected data from 111 patients with resectable periampullary lesions undergoing non-stented PPPD-PG between January 1997 and February 2003 were analyzed. Severity of postoperative steatorrhea was assessed. Surgical morbidity and mortality were evaluated. RESULTS: Complications occurred in 38 (34.2%) patients, leading to 2 (1.8%) deaths. However, neither of the deaths were related to operation. The most common complication was gastric atonia (14.4%). Pancreatic leakage occurred only in 1 (0.9%) patient. Overall, there was no steatorrhea in 92 (82.8%) patients after non-stented PPPD-PG, including 69 (62.2%) patients without replacement of pancreatic enzymes and 23 (20.7%) patients after replacement of pancreatic enzymes. Moderate steatorrhea occurred in 17 (15.3%) patients, and severe steatorrhea only in 1 (0.9%) patient. Steatorrhea was significantly correlated with the consistency of pancreatic parenchyma (P=0.037), instead of the diameter of pancreatic duct. CONCLUSIONS: The findings of low incidence of pancreatic leakage and steatorrhea in this non-stented PPPD-PG study imply that pancreatic stenting and duct-to-mucosa anastomosis may not be crucial in prevention of pancreatic leakage and exocrine insufficiency after reconstruction with pancreaticogastrostomy. PMID- 15783044 TI - Feasibility study of imaging of intraductal papillary mucinous tumors of the pancreas based on integral photography. AB - BACKGROUND/AIMS: The principle of integral photography was first proposed by Lippman in 1908. Recently, the three-dimensional display system based on the principle of integral photography was developed. It projects three-dimensional objects using a lens array called a fly's eye lens and photographic film. Some groups have used it for surgical navigation in brain surgery. In this study, we tried to generate integral photographic images of intraductal papillary mucinous tumors of the pancreas, and evaluated the feasibility of this method. METHODOLOGY: Five patients with an intraductal papillary mucinous tumor of pancreas were studied. We used MRI data as three-dimensional data. MRI data were acquired with a 1.5-T clinical imager (Signal.5; GE Medical Systems, U.S.A). We used multi-slab single-shot fast spin-echo sequences. Section thickness was between 2 and 3mm in the coronal plane. From these data, integral photographic images were generated on a liquid crystal display by a three-dimensional rendering algorithm. RESULTS: Three-dimensional images using the principle of integral photography could be generated in all 5 cases. We could recognize these images as three-dimensional images by slightly moving the viewpoint to up, down, left, and right. From these images, we were able to grasp the three-dimensional relationship between the tumor, bile duct, and pancreatic duct. Using these three dimensional images, a variety of minimally invasive surgical procedures were performed for intraductal papillary mucinous tumor of the pancreas, more safely and speedily than formerly. CONCLUSIONS: The images based on integral photography are suitable for medical imaging and are useful for surgical planning. PMID- 15783045 TI - Laparoscopic distal pancreatectomy for insulinoma with preservation of the spleen. AB - We report on a successful laparoscopic distal pancreatectomy due to insulinoma, preserving the spleen and the splenic vessels in a 29-year-old male patient who presented with repeated syncope due to hypoglycemia. The ultrasound exam did not show the pancreatic lesion; it was only the angiotomography of the pancreas that revealed a 3-cm mass located at the transition from the body to the tail of the pancreas. The laparoscopic distal pancreatectomy was performed using a harmonic scalpel (Ethicon EndoSurgery/UltraCision), without mechanical suturing. There were no intra- or postoperative complications or hypoglycemias during the 6 months of follow-up. When it is performed by experienced laparoscopic surgeons, this is a technically feasible procedure, safe for the treatment of benign lesions of the pancreas body and tail. PMID- 15783046 TI - Successful treatment for rupture of pancreaticoduodenal artery aneurysm: two case reports. AB - We report 2 cases of ruptured pancreaticoduodenal artery aneurysm (PDAA) treated by transcatheter embolization (TAE). In the first case, a 63-year-old man complained of sudden abdominal pain and was transferred to our hospital because he collapsed in a state of shock. Abdominal computed tomography (CT) revealed retroperitoneal hematoma and ascites. Abdominal angiography showed bleeding from one of the branches of the inferior pancreaticoduodenal artery. The ruptured PDAA was terminated by TAE. In the second case, a 65-year-old man experienced sudden abdominal pain. Abdominal CT revealed a retroperitoneal hematoma. He received TAE to terminate bleeding from a PDAA, but his abdominal pain worsened. At operation, ileus caused by the hematoma compressing the transverse colon was diagnosed, and cecostomy was performed. He recovered well and was discharged a few days later. In summary, a patient with a ruptured PDAA should first be treated by TAE, followed if necessary by surgery. PMID- 15783047 TI - Successful treatment for ampullary submucosal bleeding-induced pancreatitis: a rare sequla of hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (Rendu-Osler-Weber disease) is clinically characterized by the triad of mucocutaneous telangiectasia, recurrent epistaxis, and a familial history. We describe a 68-year-old woman with hereditary hemorrhagic telangiectasia undergoing transduodenal papillectomy for ampullary submucosal bleeding-induced pancreatitis. When she was 63 years old, she presented with abdominal pain due to acute pancreatitis, and from the age of 68, frequency of the symptom increased a couple of times a month. Gastrofiberscopy showed enlarged ampulla of Vater. Doppler ultrasonography showed rich and tortuous signals of bloodstream at submucosal region of the ampulla. Arteriogram revealed that hypervascular staining was fed by the inferior pancreaticoduodenal artery. Microcoil embolization was attempted and hypervascular staining disappeared. However, one week later, endoscopic papillotomy was unable to be performed due to risk of uncontrollable bleeding since Doppler ultrasonography showed signals of bloodstream at submucosal region of the ampulla again. Finally, she underwent open transduodenal papillectomy. She had an uneventful postoperative course, and has been free of pancreatitis for 20 months after surgery. We also review the literature and discuss the surgical indication for gastrointestinal and hepato-biliary-pancreatic involvement with hereditary hemorrhagic telangiectasia. To our knowledge, this is the first reported case of successful surgical treatment for pancreatitis due to sequla of hereditary hemorrhagic telangiectasia. PMID- 15783048 TI - Effect of distal splenorenal shunt plus splenopancreatic disconnection on glucose and amino acid metabolism. AB - BACKGROUND/AIMS: The Warren-Zeppa distal selective splenorenal shunt (DSRS) is aimed at decompressing esophageal varices by a distal splenorenal shunt, at the same time ligaturing left gastric, gastroepiploic and umbilical veins. In the long-term follow-up the loss of shunt selectivity was observed in several cases. Therefore, Inokuti and Warren reported splenopancreatic disconnection (SPD) to prevent splenic collaterals from stealing portal venous blood. METHODOLOGY: This report presents metabolic data (K-glucose, IRI in iv-GTT, Fischer ratio) of 10 cirrhotic patients operated with DSRS plus SPD. RESULTS: K-glucose of iv-GTT was improved from 2.3 +/- 0.2 to 2.5 +/- 0.2. Fischer ratio was also improved from 1.58 to 2.0 after the operation. IRI behavior tends to be lower after operation as a result of devascularlization of the collateral circulation by the selective shunt. CONCLUSIONS: From these results it seems that DSRS+SPD has favorable effects on glucose and amino acid metabolism. PMID- 15783049 TI - Association of the number of metastatic perigastric lymph nodes with long-term survival in gastric cancer. AB - BACKGROUND/AIMS: The presence of metastases in lymph nodes is an important prognostic factor in patients with gastric cancer. Currently, the total number of metastatic lymph nodes (MLN) is thought to be more prognostically significant than the anatomical distribution of MLN. METHODOLOGY: The associations of the total number of MLN and of the number of perigastric (PG) MLN with long-term survival were retrospectively evaluated using uni- and multivariate analysis in 481 nodepositive gastric cancer patients who underwent potentially curative resections. RESULTS: The number of PG MLN was a better prognostic indicator than the total number of MLN (indices of correlation 0.2908 and 0.3424, respectively). Good surgical results were obtained in patients with 5 PG MLN or fewer. CONCLUSIONS: The number of perigastric metastatic lymph nodes is a more conveniently obtained and reliable prognostic factor than the total number of metastatic lymph nodes in patients with node-positive gastric cancer. PMID- 15783050 TI - Loss of KAI1 expression in gastric cancer. AB - BACKGROUND/AIMS: The purpose of this study was to determine whether the loss of KAI1 expression was related to the clinicopathologic features of patients with gastric cancer. METHODOLOGY: The expression of KAI1 protein was investigated retrospectively in 174 patients with gastric cancer. Immunohistochemical staining of the paraffin sections was performed using polyclonal antibody to KAI1. RESULTS: KAI1 was consistently and highly expressed in normal gastric epithelium, while 47 (27%) of 174 patient samples had KAI1-positive expression in the primary gastric carcinoma. There were no significant differences of age, gender, and tumor location. With regard to the clinicopathological characteristics, significant differences were observed in histological types (p<0.001), increased tumor stage (p<0.001), lymph node (p<0.001) and distant metastasis (p<0.001). KAI1-negative patients had a worse prognosis than that of positive patients in overall survival, but loss of KAI1 expression was not an independent prognostic indicator (p=0.779) by multivariable analysis. CONCLUSIONS: Loss of KAI1 of expression is likely to predict metastasis and poor clinical outcome in gastric cancer patients. For the purpose of predicting the prognosis of gastric cancer patients, it is important to discriminate whether the carcinoma cells have loss of KAI1 expression. PMID- 15783051 TI - Type III intestinal metaplasia, Helicobacter pylori infection and gastric carcinoma risk index in an Italian series of 1750 patients. AB - BACKGROUND/AIMS: To evaluate the utility of 2 biopsies of antrum and gastric body on routine endoscopy for the assessment of type III intestinal metaplasia (IM-3) and Helicobacter pylori (Hp), 1750 patients (pts) (895 males; 855 females) were considered from June'98 to June'00. METHODOLOGY: Specimens were graded 0 to 3 for atrophy, IM-3 and Hp. 610 pts treated previously with antibiotics or not eligible for biopsy were excluded from initial 2360 pts. RESULTS: IM-3 was found in 118 pts (6.7%), 100 pts (5.7%) only in the antrum. 10 of 355 pts (2.8%) with normal endoscopy and 47 of 702 (6.6%) with non-erosive endoscopic gastritis resulted IM 3 positive in the antrum. 709 pts (40.5%) were positive for Hp in antrum and/or corpus. The presence of Hp and IM-3 in the antrum was not correlated (p=0.99; Spearman test). A positive correlation (p=0.000) between duodenal ulcer and Hp was found when antral Hp positivity was taken into account. The gastric carcinoma risk index (GCRI) was found in 358 pts (20.4%); in this group 131 pts (36.6%) were Hp positive, 81 pts (22.65%) had IM-3 only in the antrum, 184 pts (51.4%) had atrophy. CONCLUSIONS: The incidence of IM-3 is low (6.7%) in routine endoscopy. Normal endoscopy doesn't exclude the presence of IM-3. Biopsy is necessary to discover IM-3 in the antrum in 5.3% of pts with normal or aspecific endoscopic gastritis. Application of the GCRI might be useful for identifying a group of patients carrying a higher risk for gastric carcinoma. PMID- 15783052 TI - Induction of lymphokine-activated cytotoxic T lymphocytes stimulated by dendritic cells and autologous tumor from a patient with gastric cancer and their effects in vitro. AB - BACKGROUND/AIMS: The purpose of the study was to generate lymphokine-activated cytotoxic T lymphocytes stimulated by dendritic cells (DC) and autologous tumor from a patient with gastric cancer and to clarify their cytotoxic effects in vitro. METHODOLOGY: DC was induced by interleukin-4 (IL-4) and granulocyte macrophage-colony-stimulating factor (GM-CSF) from the peripheral blood mononuclear cells (PBMC). Then, PBMC was incubated with mitomycin C-treated tumor cells and DC, and following that was activated with IL-2 and anti-CD3. Induction of DC and cytotoxic T cells (CTL) were confirmed by the analyses of the cell surface antigens, killing activities, and blocking tests. RESULTS: Induction of DC and cytotoxic T cells (CTL) was confirmed by the analyses of the cell surface antigens, killing activities, and blocking tests. In vitro study demonstrated that lymphokine-activated lymphocytes pulsed by DCs and autologous tumor contained the largest population of CTLs, the greatest production of IFN-gamma, and the greatest ATK activity. CONCLUSIONS: Those results indicated that CTLs could be generated in vitro from a patient with gastric cancer more successfully by this method than by conventional methods, suggesting the possibility of a new immunotherapy for the treatment of gastric cancer. PMID- 15783053 TI - Laparoscopy-assisted and open distal gastrectomies for early gastric cancer at a general hospital in Japan. AB - BACKGROUND/AIMS: At general hospitals in Japan, laparoscopic surgery for early gastric cancer is not yet popular. The benefits and feasibility of this procedure remain to be established. The aim of this study was to evaluate the surgical outcome of laparoscopy-assisted distal gastrectomy (LADG) in comparison with open distal gastrectomy (ODG) in a general hospital. METHODOLOGY: We performed LADG in 20 patients with early gastric cancer between 2000 and 2001. Clinicopathologic data, blood analyses, clinical course and financial cost of treating patients with LADG were compared with 22 patients treated with ODG between 1998 and 1999. RESULTS: All patients were treated successfully by LADG. Neither reduced operative curability nor increased complications were found with this procedure. Although LADG required a significantly longer operation time than ODG, blood loss was lower in LADG than in ODG. The leukocyte count on day 1 and day 3, and serum C-reactive protein levels on day 1 were significantly lower after LADG than after ODG. There was no significant difference between LADG and ODG in the period and volume of analgesics required. High body temperature continued longer after ODG than after LADG. The first walking, passage of flatus and oral diet initiation were significantly earlier in patients with LADG than in those with ODG. LADG required a significantly shorter hospital stay and less total hospital charge than ODG. CONCLUSIONS: Laparoscopy-assisted distal gastrectomy offered faster recovery of gastrointestinal function, a shorter hospital stay, and consequently less financial cost when compared with open surgery. Therefore, LADG may be a safe and recommendable procedure for patients with early gastric cancer at general hospitals in Japan. PMID- 15783054 TI - Posterior gastric artery in angiograms and its surgical importance. AB - BACKGROUND/AIMS: The incidence and the clinical importance of the posterior gastric artery (PGA) have not been clarified. In Japan, as in the West, the incidence of adenocarcinoma of the upper stomach has increased recently. The purpose of this study was to clarify the features of the posterior gastric artery by analyzing a large series of digital subtraction angiography, and to determine its clinical significance. METHODOLOGY: Three hundred and fourteen patients who underwent angiography between 1991 and 2001 at the Kochi Medical School were studied. The length from the root of the splenic artery and the internal diameter of PGA were investigated celiacographically. The correlation of the body mass index (BMI) and the features of PGA were evaluated. RESULTS: PGA was visualized in 175 patients (55.7%) celiacographically. PGA averaged 3.2-7.4cm (mean 5.4) in length from the root of the splenic artery and the size was 0.7-2.1mm (mean 1.4). The rate of depiction in patients with liver cirrhosis (73.2%) was significantly higher than that in those without (30.5%). No significant difference in length or size of PGA was seen by gender, age, or hepatitis viral infection. No correlation was found between the length or size of PGA and BMI. CONCLUSIONS: Our findings show that the anatomical features of PGA can be visualized by clinical methods useful in planning surgical treatment. PMID- 15783055 TI - Total gastrectomy for gastric carcinoma. AB - BACKGROUND/AIMS: Gastric cancer is one of the most common organ cancers all around the world and surgical resection is essential for treatment. Total gastrectomy is the procedure of choice for treatment of proximal gastric cancer. Mortality and morbidity risks of this procedure are high, especially among the elderly. METHODOLOGY: Thirty-eight gastric cancer patients underwent total gastrectomy in the Third Surgical Clinic of Izmir Ataturk Training and Research Hospital between 1996 and 2001. Age, gender, location of the tumor, histopathological findings, TNM stage, type of anastomosis, operation time, blood transfusions, oral food intake, postoperative hospital stay, morbidity, mortality both early and late, and survival rate were evaluated. RESULTS: Mean age of the patients was 59.5 years (22-85 years). Sites of the tumors were: cardia 28.9%, cardia and corpus 15.8%, corpus 34.3%, corpus and antrum 18.4%, linitis plastica 2.6%. Histological types were adenocarcinoma (97.4%), and squamous cell carcinoma (2.6%). TNM stages were: stage la 2.6%, stage II 7.9%, stage IIIa 39.5%, stage IIIb 42.1%, and stage IV 7.9%. Esophagojejunostomies were performed manually (34.3%) or by circular staplers (65.7%). Operation time ranged between 3 and 6.5 hours. Gastric tubes were removed on the fourth postoperative day. Average postoperative hospital stay was 12.9 days. Postoperative morbidity was 21%. Wound infection occurred in three patients (7.9%), pulmonary infection occurred in two patients (5.2%) and anastomotic stricture developed in three patients (7.9%). Hospital mortality was 20%. Anastomotic leak occurred in five cases (13.2%) and all died on days 8, 13, 14, 26, and 30. Three patients (7.9%) died of cardiac complications on days 1, 5, and 29. Twelve patients survived for less than one year and eight patients survived for one to two years. Average survival was 10.5 months for 20 out of 30 patients (median 8 months). Ten patients are still alive after 14.3 months. Four patients have been living for less than one year, another four patients for one to two years, and two patients for more than two years. CONCLUSIONS: Total gastrectomy, either performed with a curative or palliative aim, is a safe procedure with acceptable mortality rates. 89.5% of our cases were stage III or IV resulting in a low survival rate. Longer survival rates can be achieved in patients with comparatively earlier stages. PMID- 15783056 TI - Delayed gastric emptying after distal gastrectomy for gastric cancer. AB - BACKGROUND/AIMS: Gastric surgery may potentiate delayed gastric emptying. The aims of the study were to determine the frequency and causes of delayed gastric emptying in patients who had undergone distal gastrectomy for gastric carcinoma and to assess which factors predispose to its development. METHODOLOGY: 209 patients, who had undergone gastrectomy, were evaluated. RESULTS: Delayed gastric emptying occurred in 4.3% of patients receiving a Billroth-I, and 15.5% of patients receiving a Roux-Y reconstruction (p = 0.01). The patients who had these consecutive symptoms following Roux-Y operation were all clinically diagnosed as having Roux stasis syndrome. The delayed gastric emptying after Roux-Y operation was more frequent in patients receiving extensive lymph node dissection than those receiving conventional dissection (p<0.05). The symptoms spontaneously subsided, and postoperative body weight loss was not significant. CONCLUSIONS: In spite of the strong association between delayed gastric emptying and the Roux-Y procedure, it enables a relatively early return to oral intake. PMID- 15783057 TI - Esophageal manometric changes and gastroesophageal reflux symptoms after distal gastrectomy for gastric cancer. AB - BACKGROUND/AIMS: Gastroesophageal reflux is known to be a common complication after gastrectomy. However, its mechanism is not completely understood. We investigated the effects of distal gastrectomy for gastric cancer on the lower esophageal sphincter (LES) and esophageal motility. METHODOLOGY: In 18 patients who underwent distal gastrectomy reconstructed with Billroth I method for gastric cancer, esophageal motility and LES function were evaluated by means of a low compliance manometric system. The LES pressure was determined by a rapid pull through technique. Endoscopy before and after operation determined presence or absence of esophagitis and hiatus hernia. RESULTS: No significant differences were observed in esophageal contractile amplitudes before and after distal gastrectomy. After distal gastrectomy, five patients had reflux symptoms of heartburn and regurgitation; 11 had none. Endoscopy revealed esophagitis after distal gastrectomy in two patients with reflux symptoms and one patient without reflux symptoms. The LES pressure in patients with reflux symptoms decreased significantly after distal gastrectomy (before gastrectomy: 26.1 +/- 1.1 mmHg, after distal gastrectomy: 15.3 +/- 3.5 mmHg, p<0.05). There was no significant change in patients without reflux symptoms. CONCLUSIONS: This study demonstrated that LES pressure after distal gastrectomy in patients with reflux symptoms was significantly lower than that before gastrectomy. This result suggested that LES pressure decrease plays an important role in development of gastroesophageal reflux after distal gastrectomy reconstruction with the Billroth I method. PMID- 15783058 TI - Therapeutic strategy for scirrhous type gastric cancer. AB - BACKGROUND/AIMS: As no appropriate therapeutic strategy has yet been established in scirrhous type gastric cancer, we retrospectively analyzed the therapeutic outcomes in patients with this type of cancer. METHODOLOGY: A total of 183 patients with scirrhous type gastric cancer were enrolled in the study. 127 of them underwent resection; 61 potentially curative gastrectomy; 66 palliative resection; and 56 had no surgery. RESULTS: Univariate analysis revealed that the number of metastatic lymph nodes and the depth of invasion influenced prognosis in curatively resected cases, whereas no factor did so after palliative resection. Multivariate analysis showed that prognosis was affected independently by peritoneal metastasis and non-regional lymph node metastasis in all resected cases, but by the number of metastatic lymph nodes in curatively resected cases. There was no significant difference in survival between patients undergoing and those not undergoing palliative gastrectomy. Prophylactic (6) and therapeutic CHPP (12) had no efficacy on peritoneal metastasis. Furthermore, left upper abdominal evisceration (LUAE) (9) did not improve long-term results in curatively resected cases. CONCLUSIONS: In scirrhous type gastric cancer, gastrectomy including extended lymph node dissection is justified only in patients with limited lymph node metastasis, and palliative gastrectomy should be not performed because it has no efficacy on survival. PMID- 15783059 TI - A case of gastric hyperplastic polyp with malignant transformation. AB - In this paper we report a case of hyperplastic polyp with malignant transformation. The patient was followed up by annual radiographic and endoscopic examination during 9 years. The first gastroscopy revealed a semi-pedunculated polyp beneath the esophageal-cardiac junction, 1.5cm in diameter and slightly reddish with smooth surface. Histological findings showed a hyperplastic polyp. There was slight change in size, but no change in histological examination of the lesion during the follow-up. In 1996, the biopsy specimen showed an atypical epithelium and the polyp was removed by snear polypectomy. A microscopic examination revealed a well-differentiated adenocarcinoma. Immunohistochemically, p53 and Ki-67 immunostaining showed positive in the carcinoma portion of the adenocarcinoma bearing a hyperplastic polyp in the stomach. PMID- 15783060 TI - Advanced gastric cancer with peritoneal dissemination successfully treated with paclitaxel and doxifluridine: a case report. AB - A 50-year-old female presenting with severe ascites and anemia and diagnosed with advanced gastric cancer was admitted to our hospital. Endoscopic examination revealed an edematous lesion with redness and a giant fold in the stomach with poor expansion. The histological examination of biopsy specimens from the edematous lesion revealed signet-ring-cell carcinoma. Computed tomography demonstrated a thickening of the gastric wall, severe ascites, and peritoneal dissemination in the Douglas pouch. Paclitaxel (70mg/m2) was administered to the patient on days 1, 8, and 15, with doxifluridine (533mg/m2) for five days per week, on a 28-day cycle. By completion of the first course of treatment, the ascites had disappeared, the tumor in the Douglas pouch had shrunk, and the thickening of the gastric wall had lessened. In addition, the fold in the stomach appeared by endoscopic examination to have resumed its normal thickness, no malignant cells were detected in a biopsy, and the thymidine phosphorylase activity in the tumor tissue was two-fold greater than that before chemotherapy. After three treatment courses, the number of apoptotic cells had apparently increased compared with the prechemotherapy number. The only adverse drug reactions that were observed were grade 2 alopecia and grade 1 myalgia. After thirteen courses of chemotherapy over the past one year, both primary and metastatic lesions seem to be regressing. This case study suggests that paclitaxel plus doxifluridine therapy is effective and well-tolerated in non resectable gastric cancer patients. PMID- 15783061 TI - Third-line chemotherapy with paclitaxel, irinotecan hydrochloride and cisplatin for recurrent gastric cancer: a case report. AB - With the availability of new chemotherapeutic agents such as S-1 and paclitaxel (TXL) for advanced gastric cancer, the development of a strategy for a third-line chemotherapy is urgently needed. We treated a patient with recurrent gastric cancer using TXL, irinotecan hydrochloride (CPT-11) and cisplatin (CDDP) as a third-line chemotherapy. The patient was a 46-year-old man who had undergone total gastrectomy for advanced gastric cancer with lymph node metastases. For postoperative recurrence, he was first treated with S-1 as an outpatient; however, tumor markers increased, and para-aortic lymph node metastasis was revealed by thoracic and abdominal CT scan. A second-line therapy with weekly TXL and CDDP was then added, but resulted in PD. Therefore, combination chemotherapy with TXL, CPT-11 and CDDP was started biweekly as a third-line chemotherapy. TXL (80mg/m2) was infused over 1 hour after short premedication, followed by CPT-11 (25mg/m2) and CDDP (15mg/m2) over 30 min. After 6 courses of this therapy, the serum AFP and TPA returned to normal, and the size of the metastatic para-aortic lymph nodes reduced. The effect of this therapy was judged as PR and the toxicity of this regimen was tolerable. The patient has undergone 10 courses of this therapy and is maintaining a clinical PR. The patient was able to resume his full social activities. TXL, CPT-11 and CDDP combination chemotherapy may be useful and safe for patients with recurrent gastric cancer, even after first-or second line therapy with S-1 or taxanes. PMID- 15783062 TI - Pharmaceutical sciences, scientists, and journals--a tribute to pharmaceutical research for its coming of age. PMID- 15783063 TI - Bioadhesive dosage forms for esophageal drug delivery. AB - The esophagus as a site for drug delivery has been much overlooked in comparison to the remainder of the gastrointestinal tract. The low permeability and transient nature of the esophagus means that it is unsuitable for delivery of drugs for systemic action. However, esophageal disorders including fungal infection, cancers, motility dysfunction, and damage due to gastric reflux may be treated using locally acting agents that offer benefits of reduced dosage and decreased side effects. Bioadhesive dosage forms that adhere to the esophageal mucosa and prolong contact have been investigated to improve the efficacy of locally acting agents. The rationale for local esophageal drug delivery and its limitations, the factors that determine adhesion to this organ, and the experimental models used in esophageal drug delivery research are reviewed. PMID- 15783064 TI - Oral dosage form performance tests: new dissolution approaches. AB - The performance test is one of a series of tests that compose the specification in a United States Pharmacopeia (USP) dosage form monograph. For an orally administered, nonsolution dosage form, it is usually satisfied by either a dissolution or disintegration procedure. Dissolution acceptance criteria are usually set in private negotiations between an applicant and a regulatory agency. With information about this private agreement and other information provided in a sponsor's Request for Revision to USP, the USP's Council of Experts elaborates a public dosage form monograph. Based on the relationship between the regulatory decisions and the Request for Revision, the USP dissolution procedure links to a regulatory judgment about bioavailability and bioequivalence and, ultimately, to a judgment about safety and efficacy. The current dissolution procedure and acceptance criteria are perceived as having worked well over the years and are generally accepted. This article discusses new approaches that merit consideration. These approaches focus on a) explicit use of hypothesis testing, b) use of parametric tolerance intervals, c) improved ways to set dissolution acceptance criteria, and d) a more flexible protocol to assess conformity. Application of the proposed approaches may better assess, manage, and communicate both manufacturer and consumer risk for dissolution testing. PMID- 15783065 TI - pH-dependent dissolution in vitro and absorption in vivo of weakly basic drugs: development of a canine model. AB - PURPOSE: The aim of this research was to develop a pH-dependent canine absorption model for studying pH effect on both dissolution in vitro and pharmacokinetics in vivo using the weak bases ketoconazole and dipyridamole as model drugs. METHODS: Ketoconazole and dipyridamole pH-dependent dissolution profiles in vitro were determined by dissolution test at different pH values using USP apparatus II and an Opt-Diss Fiber Optic UV System. In vivo absorption studies for ketoconazole and dipyridamole were performed with crossover design in three groups of beagle dogs under control (no treatment), pentagastrin, and famotidine treatments. Ketoconazole and dipyridamole plasma concentrations were quantified by gradient high performance liquid chromatography mass spectroscopy (HPLC MS/MS). Pharmacokinetic parameters were determined from individual plasma concentration vs. time profiles. RESULTS: Ketoconazole and dipyridamole displayed pH-dependent dissolution. Increasing the pH of the dissolution medium from 1.2 to 6.8 reduced the extent of dissolution of ketoconazole and dipyridamole at 1 h by 96% and 92%, respectively. In vivo studies in dogs under control (no treatment), pentagastrin, and famotidine treatments show marked differences in systemic ketoconazole and dipyridamole exposure. Area under the concentration-time curve (AUC) increased more than 4-fold as compared to control group, whereas it increased nearly 30 fold for ketoconazole and 9-fold for dipyridamole with pentagastrin (gastric pH approximately 2-3) as compared to famotidine (gastric pH approximately 5-7.5) treatment. CONCLUSIONS: This work demonstrates a pH-dependent dissolution in vitro and absorption in vivo for the weak bases ketoconazole and dipyridamole independent of food effects. This model is useful to examine pH-dependent effects on oral drug absorption and for screening formulations to overcome the pH dependency. PMID- 15783066 TI - FK506 microparticles mitigate experimental colitis with minor renal calcineurin suppression. AB - PURPOSE: FK506 microparticles providing selective colonic drug delivery were tested for their efficiency in a local treatment to the inflamed gut tissue in inflammatory bowel disease (IBD). Because FK506 proved its distinct mitigating potential in the treatment of IBD, risking, however, severe adverse effects, a more selective delivery to the site of inflammation may further improve efficiency and tolerability. METHODS: A model colitis was induced to male Wistar rats by trinitrobenzenesulfonic acid. FK506 was entrapped into microspheres (MS) prepared with the pH-sensitive polymer Eudragit P-4135F in order to allow drug delivery to the colon. Clinical activity score, colon/body weight index, and myeloperoxidase activity were determined to assess the inflammation, and adverse effects of FK506 resulting from its systemic absorption were quantified as well. RESULTS: The clinical activity score and myeloperoxidase activity decreased after the administration of all FK506-containing formulations. The MS formulations proved to be as efficient in mitigating the experimental colitis as the subcutaneous drug solution (myeloperoxidase activity, MS: 9.64+/-6.6 U/mg tissue; subcutaneous: 7.48+/-6.96 U/mg) and to be superior to drug solution given by oral route (oral: 12.66+/-5.46 U/mg; untreated colitis control: 21.88+/-4.12 U/mg). The FK506 subcutaneous group exhibited increased levels of adverse effects, whereas the FK506-MS group proved its potential to retain the drug from systemic absorption as evidenced by reduced nephrotoxicity. CONCLUSIONS: The development of this selective delivery system for FK506 should be given particular consideration in the treatment of IBD, as it allows therapy that profits from FK506's high immune suppressive effect with a simultaneously reduced nephrotoxicity. PMID- 15783067 TI - A polymeric nanoparticle consisting of mPEG-PLA-Toco and PLMA-COONa as a drug carrier: improvements in cellular uptake and biodistribution. AB - PURPOSE: To evaluate a new polymeric nanoparticulate drug delivery formulation that consists of two components: i) an amphiphilic diblock copolymer having tocopherol moiety at the end of the hydrophobic block in which the hydrophobic tocopherol moiety increases stability of hydrophobic core of the nanoparticle in aqueous medium; and ii) a biodegradable copolyester having carboxylate end group that is capable of forming ionic complex with positively charged compounds such as doxorubicin. METHODS: A doxourubicin-loaded polymeric nanoparticle (Dox-PNP) was prepared by solvent evaporation method. The entrapment efficiency, size distribution, and in vitro release profile at various pH conditions were characterized. In vitro cellular uptake was investigated by confocal microscopy, flow cytometry, and MTT assay using drug-sensitive and drug-resistant cell lines. Pharmacokinetics and biodistribution were evaluated in rats and tumor-bearing mice. RESULTS: Doxorubicin (Dox) was efficiently loaded into the PNP (higher than 95% of entrapment efficiency), and the diameter of Dox-PNP was in the range 20-25 nm with a narrow size distribution. In Vitro study showed that Dox-PNP exhibited higher cellular uptake into both human breast cancer cell (MCF-7) and human uterine cancer cell (MES-SA) than free doxorubicin solution (Free-Dox), especially into drug-resistant cells (MCF-7/ADR and MES-SA/Dx-5). In pharmacokinetics and tissue distribution study, the bioavailability of Dox-PNP calculated from the area under the blood concentration-time curve (AUC) was 69.8 times higher than that of Free-Dox in rats, and Dox-PNP exhibited 2 times higher bioavailability in tumor tissue of tumor-bearing mice. CONCLUSIONS: Dox-PNP exhibited enhanced cellular uptake of the drug. In the cytotoxic activity study, this improved cellular uptake was proved to be more advantageous in drug resistant cell. Dox-PNP exhibited much higher bioavailability in blood plasma and more drug accumulation in tumor tissue than conventional doxorubicin formulation. The results of this study suggest that the PNP system is an advantageous carrier for drug delivery. PMID- 15783069 TI - Reversible lipidization prolongs the pharmacological effect, plasma duration, and liver retention of octreotide. AB - PURPOSE: Octreotide (OCT) was reversibly lipidized to improve the pharmacological effect and to increase the plasma half-life and the liver retention of OCT for greater therapeutic potential in the treatment of liver cancers such as hepatocellular carcinoma. METHODS: OCT was chemically modified using reversible aqueous lipidization (REAL) technology. REAL-modified OCT (REAL-OCT) was characterized with high performance liquid chromatography (HPLC) and matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. A single dose of OCT or REAL-OCT or vehicle only was subcutaneously administered to male Sprague-Dawley rats, and the plasma growth hormone (GH) levels were measured after an intravenous injection of 2.5 microg/kg of growth hormone releasing factor (GRF) to assess the ability of REAL-OCT on GH inhibition. Radio-iodinated Tyr3-OCT (TOC) and REAL-TOC were used for pharmacokinetic studies. RESULTS: At 0.1 mg/kg, REAL-OCT inhibited the GRF induced GH surge in rats for a greater than 24-h period in comparison to the 6-h period for OCT. The distribution and elimination half-life for 125I-REAL-TOC were 1.4 h and 6.6 h, respectively, which were significantly longer than those of 125I TOC. Sustained high blood concentrations and reduced in vivo degradation were observed for 125I-REAL-TOC. In addition, 125I-REAL-TOC appeared to be targeted to the liver with persistent high liver retention. CONCLUSIONS: REAL-OCT has a significantly enhanced pharmacological effect, and this is most likely due to the favorable changes in the pharmacokinetic parameters upon lipidization. The observed liver targeting effect of REAL-TOC suggests that REAL-OCT might be advantageous over OCT in treating liver cancers. PMID- 15783068 TI - Engineering polysaccharide-based polymeric micelles to enhance permeability of cyclosporin A across Caco-2 cells. AB - PURPOSE: To assess and compare the effectiveness of two types of polysaccharide based micelles as delivery vehicles for poorly water soluble drugs by monitoring their permeability across Caco-2 cell monolayers. METHODS: Dextran (DEX) and hydroxypropylcellulose (HPC) were hydrophobically modified (HM) by grafting polyoxyethylene cetyl ether (POE-C16, 15 mol% and 5.4 mol%, respectively). The onset of micellization and mean diameter of polymeric micelles formed by HM-DEX and HM-HPC were determined by fluorescence spectroscopy and dynamic light scattering, respectively. Cyclosporin A (CsA)-loaded polymeric micelles were prepared by a dialysis procedure, and the amount of incorporated CsA was assayed by high performance liquid chromatography (HPLC). The stability of micelles in simulated gastric and intestinal fluids was studied as a function of contact time, and their cytotoxicity toward Caco-2 cells was evaluated using the MTT colorimetric assay. The bidirectional transport across Caco-2 cell monolayers of CsA entrapped in HM-DEX and HM-HPC micelles and of the polymers themselves was evaluated in the presence and absence of P-glycoprotein inhibitor. RESULTS: The amount of CsA incorporated in HM-HPC and HM-DEX micelles reached 5.5 and 8.5% w/w, respectively (entrapment efficiency of 22% or more). The polymeric micelles exhibited high stability in gastric and intestinal fluids and no significant cytotoxicity toward Caco-2 cells. The apical to basal permeability of CsA across Caco-2 cells increased significantly when loaded in polymeric micelles compared to free CsA. CONCLUSIONS: Polysaccharide-based polymeric micelles are promising carriers for the oral delivery of poorly water soluble drugs. In vitro tests indicate that, overall, HM-HPC micelles are more effective compared to HM-DEX micelles. PMID- 15783070 TI - Altered pharmacokinetics of paclitaxel in experimental hepatic or renal failure. AB - PURPOSE: The aim of this study was to investigate the effect of hepatic or renal insufficiency on the pharmacokinetics of paclitaxel in rats. METHODS: Rats were treated with carbon tetrachloride (CCl4; 0.5 ml/kg) to induce hepatic failure or were subjected to 5/6 nephrectomy (5/6 Nx) to induce renal failure. Paclitaxel (3 mg/kg) was administered intravenously or intraportally. Testosterone 6beta hydroxylase activity, which is a marker of CYP3A activity, was measured in rat liver microsomes from CCl4-treated or 5/6 Nx rats. RESULTS: After paclitaxel was administered intravenously, total body clearance was significantly reduced by 73% and 34% relative to each control value in CCl4-treated and 5/6 Nx rats, respectively (control, 1.82+/-0.42 vs. CCl4-treated, 0.49+/-0.11; sham, 1.54+/ 0.07 vs. 5/6 Nx, 1.01+/-0.12 L h(-1) kg(-1); mean+/-SE, n = 5 to 6). Testosterone 6beta-hydroxylase activity was reduced by 92% and 59% relative to each control value in rat liver microsomes from CCl4-treated and 5/6 Nx rats, respectively. After the intraportal administration of paclitaxel, apparent clearance was reduced by 85% relative to control value in rats with hepatic failure, while that in rats with renal failure was the same as the reduction in systemic clearance. CONCLUSIONS: These results suggested that not only hepatic failure but also renal failure could modify the pharmacokinetics of paclitaxel in vivo. PMID- 15783071 TI - Pharmacokinetics of substrate uptake and distribution in murine brain after nasal instillation. AB - PURPOSE: This study was conducted to develop a physiologically relevant mathematical model for describing brain uptake and disposition of nasally administered substrates. METHODS: [14C]-antipyrine, [14C]-diazepam, [3H]-sucrose, or [3H]-verapamil was administered nasally to CF-1 mice. P-glycoprotein (P-gp) deficient mice also received [3H]-verapamil to probe the influence of P-gp on uptake/distribution. Mice were sacrificed at selected intervals, and 20 serial 300-microm coronal brain sections were obtained to determine radioactivity. A series of compartmental pharmacokinetic models was developed and fit to concentration vs. time/distance data. RESULTS: After nasal instillation, substrate concentration was highest in the olfactory bulb and decreased with distance. In the absence of transport-mediated flux, peak brain exposure occurred at 6 h. A catenary pharmacokinetic model with slice-specific brain-to-blood efflux rate constants and slice-to-slice diffusivity factors was capable of fitting the data. P-gp limited fractional absorption of [3H]-verapamil via efflux from the nasal cavity and olfactory epithelium. P-gp also increased the rate constants associated with [3H]-verapamil efflux 1.5- to 190-fold, depending on brain region. P-gp limited [3H]-verapamil uptake from the nasal cavity into brain and facilitated removal of [3H]-verapamil from brain during rostral-to-caudal distribution. CONCLUSIONS: Taken together, the data and associated modeling provide a comprehensive assessment of the influence of P-gp on brain uptake and disposition of nasally administered substrates. PMID- 15783072 TI - A randomly coiled, high-molecular-weight polypeptide exhibits increased paracellular diffusion in vitro and in situ relative to the highly ordered alpha helix conformer. AB - PURPOSE: The current investigation was conducted to examine the effect of secondary structure of model polypeptides on their hindered paracellular diffusion. METHODS: Poly-D-glutamic acid (PDGlu) was selected as one of the model polypeptides because of its ability to form two secondary structures; a negatively charged random coil and an alpha-helix with partial negative charge at pH 7.4 and 4.7, respectively. Poly-D-lysine (PDL) was selected as a positively charged random coil conformation at pH 7.4. Transport experiments were conducted across both a Caco-2 cell monolayer and the intestinal membrane of Sprague-Dawley rats. Additionally, using NMR, an estimation for the diffusion coefficient and the equivalent hydrodynamic radius for each model polypeptide was obtained. RESULTS: PDGlu in the randomly coiled conformation exhibited greater paracellular transport when compared to either the same polypeptide having an alpha-helix secondary structure or the positively charged, randomly coiled PDL. CONCLUSIONS: Randomly coiled PDGlu was able to permeate through the negatively charged tight junctions of both biological membranes to a greater extent than PDGlu having an alpha-helix structure and suggests that molecular flexibility associated with the random coil conformation may play a more important role than overall charge and hydrodynamic radius on its hindered paracellular diffusion. PMID- 15783073 TI - Metformin transport by renal basolateral organic cation transporter hOCT2. AB - PURPOSE: Metformin, an antihyperglycemic agent, is eliminated by tubular secretion in addition to glomerular filtration in the human kidney. This study was performed to characterize metformin transport by human organic cation transporter 2 (hOCT2), the most abundant organic cation transporter in the basolateral membranes of the human kidney. METHODS: Accumulation of [14C]metformin was assessed by the tracer experiments in the human embryonic kidney (HEK293) cells expressing hOCT2. RESULTS: The transport of [14C]metformin was markedly stimulated in hOCT2-expressing cells compared with the vector transfected cells. The accumulation of [14C]metformin was concentrative and was dependent on the membrane potential, showing consistency with the characteristics of hOCT2. The apparent Km and Vmax values of [14C]metformin transport by hOCT2 expressing HEK293 cells were 1.38+/-0.21 mM and 11.9+/-1.5 nmol mg protein(-1) min(-1), respectively. The order of the potencies of unlabeled biguanides to inhibit [14C]metformin transport by hOCT2 was phenformin > buformin > metformin. Furthermore, [14C]metformin transport was inhibited slightly or moderately by cationic drugs such as procainamide and quinidine at respective therapeutic concentrations. CONCLUSIONS: Metformin is transported by the basolateral organic cation transporter hOCT2 in the human kidney. hOCT2 could play a role in the drug interactions between metformin and some cationic drugs. PMID- 15783074 TI - Estimation of the three-dimensional pharmacophore of ligands for rat multidrug resistance-associated protein 2 using ligand-based drug design techniques. AB - PURPOSE: Multidrug-resistance-associated protein 2 (Mrp2) shows a broad substrate specificity toward amphiphilic organic anions. This study identified key functional groups of ligand molecules for binding to rat Mrp2, determined their relative locations, and examined substrate specificity through receptor mapping using three-dimensional (3D) quantitative structure-activity relationship (3D QSAR) analysis. METHODS: Ligand-binding conformations were estimated using conformational analysis (CAMDAS) and molecular superposition (SUPERPOSE) methods to clarify the substrate specificity of rat Mrp2 in relation to 3D ligand structures. RESULTS: Two types of binding conformations of ligands for rat Mrp2 were identified. 3D-QSAR comparative molecular-field analysis (CoMFA) revealed a statistically significant model for one type, in which the steric, electrostatic, and log P contributions to the binding affinity for rat Mrp2 were 63.0%, 33.4%, and 3.6%, respectively (n = 16, q2 = 0.59, n = 3, r2 = 0.99, and s = 0.08). CONCLUSIONS: The 3D pharmacophore of ligands for rat Mrp2, and the ligand-binding region of rat Mrp2, were estimated. Ligand recognition of rat Mrp2 is achieved through interactions in two hydrophobic and two electrostatically positive sites (primary binding sites). The broad substrate specificity of rat Mrp2 might result from the combination of secondary (two electrostatically positive and two electrostatically negative sites) and primary binding sites. PMID- 15783075 TI - The relation between granule size, granule stickiness, and torque in the high shear granulation process. AB - PURPOSE: To investigate the background of the observed relationship between measured torque and granule size in high-shear granulation processes. METHODS: Torque was measured during the granulation process; the behavior of individual wet granules during compaction was investigated using micromanipulation. Surface properties of wet granules were manipulated by coating them with talc. RESULTS: The torque-granule size relationship could not be explained by the rise in mass of the individual granules; it occurs rather through an increase in stickiness of the granules when the moisture content is increased. Obviously, the increased stickiness that causes the granules to grow also increases the torque. Increased stickiness was shown to be the result of an increased deformability of the granules at higher moisture contents, in combination with a change in surface properties. The elastic-plastic behavior (ratio of elastic to plastic deformation) was found to change at increasing moisture contents. CONCLUSIONS: Our results imply that changes in the stickiness of the granular material that may be caused by changes in composition shift the torque-size relationship. This may be of particular importance when, for example, granulation results from placebo batches are used to predict the granule size of drug-containing batches. PMID- 15783076 TI - Preparation of microparticles by micromixers: characterization of oil/water process and prediction of particle size. AB - PURPOSE: A descriptive model for microparticle preparation by micromixers was developed to allow prediction of nascent microsphere size and provide a better understanding of a microscale oil/water (O/W) emulsion process. METHODS: Nascent blank microparticles were prepared by an O/W emulsion method using a micromixer. Seven dimensionless groups were derived from the relevant process parameters. A multiple linear regression model was established on an empirical basis to describe the relationship between the key process parameters and the resulting Sauter particle diameter. RESULTS: The investigated micromixer is particularly suitable for processing of low-viscosity systems. The particle size is mainly controlled by flow velocity. Reynolds number and the viscosity ratio were found to be the most important dimensionless groups regarding the preparation procedure. Particle size was predicted with an accuracy up to 100% applying the empirically derived equations. CONCLUSIONS: An O/W process using micromixers for microparticle preparation with a multitude of influencing parameters was successfully characterized by application of dimensional analysis. Dimensionless groups turned out to be suitable for prediction of microparticle size with high precision. PMID- 15783077 TI - Influence of lipid nanocapsules composition on their aptness to freeze-drying. AB - PURPOSE: To link the aptness to freeze-drying and the stability under storage to the topology of lipid nanocapsules. METHODS: The aptness to freeze-drying and the stability under storage of lipid nanocapsules prepared from different compositions with a lecithin content in the 2-20% range were estimated from the preservation of the physical structure, preventing the leakage of the oily phase. The influence of the outer shell composition and of the physical characteristics (investigated by photon correlation spectroscopy and differential scanning calorimetry) on the physical stability was correlated to the topology of the nanoparticulate carrier. RESULTS: Confirming the assumption that lecithin confers hardness to the outer shell of lipid nanocapsules, this study shows that the aptness to freeze-drying and the stability under storage depend on the Solutol/lecithin (S/L) ratio in the formulation. The DSC study points out a complexation between lecithin and trehalose, the cryoprotectant, reinforcing the stabilising properties of lecithin. CONCLUSIONS: This paper is a contribution to methodological development of the formulation of lipid nanocapsules, with a special emphasis on the aptness to freeze-drying and the stability under storage. PMID- 15783078 TI - Study of pharmaceutical solid dispersions by microthermal analysis. AB - PURPOSE: The main objective of the study was to determine the ability of microthermal analysis (microTA) to assess the crystallinity of the drug in two noncommercial pharmaceutical solid dispersions. METHODS: Pure substances, physical mixes, and solid dispersions were analyzed by microTA. The thermal values obtained by microTA were compared with data obtained by more conventional techniques like differential scanning calorimetry. RESULTS: microTA was able to detect the drug in the waxy matrix. The technique was capable of showing that relatively large amorphous drug domains (up to 70 microm) are formed during the solidification of the solid dispersion. These amorphous domains are thermodynamically unstable and can crystallize upon aging. CONCLUSIONS: microTA was successfully applied to the physical characterization of solid dispersions. Local thermal analysis (LTA) offers a unique way to probe a selected area on the surface of the sample. It is capable of melting the drug locally without melting the lower melting point excipient. These results gave an understanding of the poor dissolution performance of these solid dispersions upon aging. PMID- 15783079 TI - Raffinose crystallization during freeze-drying and its impact on recovery of protein activity. AB - PURPOSE: To study i) phase transitions in raffinose solution in the frozen state and during freeze-drying and ii) evaluate the impact of raffinose crystallization on the recovery of protein activity in reconstituted lyophiles. METHODS: X-ray powder diffractometry (XRD) and differential scanning calorimetry (DSC) were used to study the frozen aqueous solutions of raffinose pentahydrate. Phase transitions during primary and secondary drying were monitored by simulating the entire freeze-drying process, in situ, in the sample chamber of the diffractometer. The activity of lactate dehydrogenase (LDH) in reconstituted lyophiles was determined spectrophotometrically. RESULTS: Raffinose formed a kinetically stable amorphous freeze-concentrated phase when aqueous solutions were frozen at different cooling rates. When these solutions were subjected to primary drying without annealing, raffinose remained amorphous. Raffinose crystallized as the pentahydrate when the solutions were annealed at a shelf temperature of -10 degrees C. Primary drying of these annealed systems resulted in the dehydration of raffinose pentahydrate to an amorphous phase. The phase separation of the protein from the amorphous raffinose in these two systems during freeze-drying resulted in a significant reduction in the recovery of LDH activity, even though the lyophile was amorphous. CONCLUSIONS: Annealing of frozen aqueous raffinose solutions can result in solute crystallization, possibly as the pentahydrate. The crystalline pentahydrate dehydrates during primary drying to yield an amorphous lyophile. Raffinose crystallization during freeze drying is accompanied by a significant loss of protein activity. PMID- 15783080 TI - Use of 2,2'-azobis(2-amidinopropane) dihydrochloride as a reagent tool for evaluation of oxidative stability of drugs. AB - PURPOSE: To study the oxidative degradation of drugs using a hydrophilic free radical initiator, 2,2'-Azobis(-amidinopropane) dihydrochloride (AAPH). METHODS: AAPH was used as the free radical initiator to study oxidation of three model compounds (A, B, and C), which represent different oxidizable moieties. In the solution model, the drugs and AAPH were dissolved in a mixture of acetonitrile and aqueous buffer and incubated at elevated temperatures to evaluate oxidative degradation. The effects of pH and drug-AAPH ratio on the kinetics of the reaction were evaluated for compound A. Commonly used antioxidants were also evaluated by addition to solutions of drug and AAPH. In the solid-state model, blends of drug with microcrystalline cellulose were treated with AAPH and placed at elevated temperature and humidity to evaluate solid state oxidation. RESULTS: Use of AAPH resulted in selective oxidation of the model drugs by a free radical initiated process. The scope of the technique was further investigated in detail using compound A. The rate of oxidation of compound A varied directly with the concentration of AAPH. The pseudo first-order rate constants for the oxidative degradation were calculated from the kinetic data. The antioxidants were rank ordered based on their quenching activity on the rates of AAPH initiated oxidation for compound A. The concept was extended to oxidation in solid state. CONCLUSIONS: The proposed AAPH model is useful in assessing oxidative stability of drug candidates in development. PMID- 15783081 TI - Deliquescence in binary mixtures. AB - PURPOSE: Deliquescence is a first-order phase transformation from solid to solution and occurs at a specific relative humidity (RH) that is characteristic to the solid. The goal of this research was to investigate the reduction in critical relative humidity (RH0) in binary solid mixtures of deliquescent active pharmaceutical ingredients (APIs) and deliquescent excipients. METHODS: The RH where deliquescence is induced, RH0, was measured using a gravimetric water vapor sorption balance. Values were obtained for model deliquescent APIs and excipients as well as for their binary mixtures. Model APIs included ranitidine HCl and diphenhydramine HCI, and sucrose and sodium chloride were examples of excipients investigated. To probe the thermodynamics of this phenomenon, water activity (a(w)) of various saturated solutions of API and excipient systems was measured using a water activity meter. Optical microscopy was used to observe visually the phenomenon under investigation. The Ross equation was used to estimate RH0mix, and predicted values were compared with experimental results. RESULTS: There was close agreement between RH0 measurements (single-component RH0, and RH0mix) and a(w) measurements of the corresponding saturated solutions. In addition, RH0mix values were always lower than RH0 for the API and excipients alone. In general, experimentally observed RH0mix values were higher than those predicted using the Ross equation. CONCLUSIONS: Mixtures of deliquescent API and deliquescent excipient are more hygroscopic (i.e., have lower RH0) than either the API or the excipient alone. This might have significant effects on API and drug product stability. PMID- 15783082 TI - [Gender and developmental effects on perceived stress, coping, somatic symptoms and psychological disorders among children and adolescents]. AB - The aim of this one-year longitudinal study was to investigate gender and developmental effects on perceived stress related to interpersonal stressors, coping strategies, somatic symptoms and psychological disorders. Additionally, associations of perceived stress and coping styles at the first assessment with psychological problems at the second measurement were examined. In total, N = 169 Austrian children and adolescents (ages 10 to 13 years) participated in this self report study. Compared to male children and adolescents, female counterparts scored higher on perceived stress related to interpersonal stressors and on social support but evaluated a lower amount of distraction. Moreover, girls reported higher levels of somatic symptoms and anxiety/depression. Boys showed a developmental increase in anger control problems and both genders showed developmental increases in antisocial behavior. Low levels of the emotion-focused coping strategies distraction and minimization were related longitudinally to emotional and behavioral disorders. The results support gender-adjusted stress management programs applied in the primary and secondary prevention. PMID- 15783083 TI - [Parent- and teacher-reported behavior problems of first graders]. AB - Parents information about behavior, problems and life situation of children before and after first year of school were analysed and compared with data from teacher reports. At the end of first grade 131 parents were asked about behavior problems of their children using the child behavior checklist (CBCL) and additional items about living condition of the familiy and child. Findings were compared with those of a large epidemiological study conducted one year before in the same area using the same instruments. Additional teacher-reported data about behavior problems and school achievement were included. Comparing data before and after first year of school, no significant change in average CBCL total score could be observed. From parents view oppositional behavior was the most prevalent problem. There was a decrease of social problems. Children became more independent having more contact with peers. More children have their own television and computer at the end of first grade. Prevalence of headache and perfectionism as well as symptoms of tension and irritation increased. The most important problems from teachers' view are lack of attention and concentration affecting 32% of the first graders. Compared to the parents, teachers report only few oppositional and aggressive behavior. Achievement at school and behavior problems are highly correlated. Especially attention problems go along with low achievement at school. Parents' and teachers' ratings show an average correlation of r = .28 with the highest accord for attention problems and school achievement. The results of the study contribute to the question how children deal and cope with the new situation coming to school. The findings of a high rate of attention problems at school and the close relationship between behavior problems and achievement lead to the conclusion that an early prevention of behavior problems is essential to promote school performance. PMID- 15783084 TI - [Parental relationship from the perspectives of children, adolescents and clinicians. Development and clinical validation of the Parental-Representation Screening-Questionnaire (PRSQ)]. AB - The Parental-Representation-Screening-Questionnaire (PRSQ) is a new instrument designed to asses how children and adolescents evaluate their relationship with their parents. The aim of the questionnaire is to assess those aspects in the child-parent relationship that are crucial for a psychopathological risk. Conceptually, the PRSQ is based on cognitive self theory and systemic family theory. The PRSQ consists of 8 scales (36 items) for each parent: "freedom of decision", "emotional burden by the parents", "fears / overprotection", "conflict", "hostility / indifference", "aid for the parents", "punishment", and "support from the parents''. A clinic-referred consecutive sample of 152 patients of a Berlin child and adolescent psychiatry, aged 10 to 18 years, were surveyed. Adverse educational and psychosocial family factors were assessed by clinicians according to the fifths axis of the multiaxial classification system (MAS, Dilling et al. 2000). The internal reliability for 13 of the 16 scales ranges from .90 to .72. Factor analyses confirmed the factorial independence of six of the eight scales separately for mother and father. Moreover, adolescent girls reported significantly worse relations to both parents on five of the 16 scales than boys. Furthermore we found concordant correlations with the clinician's diagnosis of adverse educational and psychosocial family situations. The present results confirm the PRSQ as an economic and discriminating instrument to assess the representation of parent-child relation for clinical and for scientific purposes. PMID- 15783085 TI - MHC class I down regulation, tumour escape from immune surveillance and design of therapeutic strategies. PMID- 15783086 TI - Expression of CD44v6 correlates with cell proliferation and cellular atypia in urothelial carcinoma cell lines 5637 and HT1197. AB - CD44 comprises a family of membrane adhesion molecules encoded by a single gene and diversified by alternative splicing and extensive posttranslational modifications. Alterations of CD44 expression patterns are linked to tumour invasion and formation of metastases. However, CD44 expression and its relation to the biological properties of tumours vary depending on the tumour type and origin. In transitional cell carcinoma of the urinary bladder, low CD44 expression is linked to enhanced tumour aggressiveness. We studied CD44 expression in two urothelial cancer cell lines, HT1197 and 5637. CD44s and a v6 variable exon-containing splice variants were detected in both cell lines by reverse transcription-PCR and by commercially available monoclonal antibodies. In both cell lines, Western blot analysis detected immunoreactive proteins with approximate sizes 70-85 kD, 95-110 kD, and 120-140 kD with CD44v6 antibody and weak bands with size 70-98 kD with CD44s antibody. At the cellular level, the pattern of CD44 immunoreactivity correlated with a lower level of cell differentiation and a higher degree of cell proliferation. In HT1197 cells, the CD44v6 was detected predominantly in small proliferating cells and in large multinuclear atypical cells. CD44s and CD44v6 displayed low immunoreactivity in HT1197 cells with a higher degree of epithelial differentiation. The 5637 cells expressed CD44v6 strongly and CD44s weakly. We conclude that CD44v6 expression correlates with a higher proliferative activity and with a stem cell-like phenotype in both cell lines and with cellular atypia in HT1197 cells. PMID- 15783087 TI - Characteristic of two mouse bcr-abl-transformed cell lines: I. General properties of the cells. AB - In an effort to develop an experimental system suitable for immunological studies in which Bcr-Abl-positive cells are to be used as antigens, we examined the properties of two mouse (Balb/c) established cell lines that express the Bcr-Abl protein and are oncogenic for syngeneic animals. Under standard conditions the two cell lines, viz. Ba-p210 (B210) and 12B1, expressed comparable amounts of the Bcr-Abl protein. However, they differed in a number of characteristics. From the morphological point of view, B210 cells were the more homogeneous, being mainly represented by leukaemic blastic cells with a large number of AgNORs as markers indicating a high proliferative activity. 12B1 cells were more polymorphic and giant cells were detected within their populations. Many 12B1 cells exhibited nuclear segmentation and "band-like" structures. Markers of proliferation were less frequent in 12B1 and the tendency for aging was more pronounced in these cells. The 12B1 cells were slightly more sensitive to imatinib mesylate than B210 cells. In B210 cells, the expression of MHC class I was downregulated, which was not the case with 12B1 cells. Both cell lines induced leukaemia-like disease in mice after intravenous application but, as compared with B210, 12B1 cells were about 100 times more oncogenic and the disease they induced was more aggressive. Moreover, 12B1, but not B210, induced tumours after subcutaneous or intraperitoneal inoculation. PMID- 15783088 TI - Immunotherapeutic efficacy of vaccines generated by fusion of dendritic cells and HPV16-associated tumour cells. AB - Utilization of vaccines generated by fusion of dendritic cells and tumour cells is a promising approach to tumour immunotherapy. We have examined the therapeutic efficacy of vaccines generated by fusion of HPV16-associated tumour cells TC-1 with syngeneic and allogeneic dendritic cells. Locally administered hybrid cells generated by fusion of MHC class I+ TC-1 cells and syngeneic DC inhibited the growth of MHC class I+ TC-1 tumours, but not the growth of MHC class I- TC-1/A9 derived tumours. The growth of TC-1 tumours was also inhibited by hybrids generated by fusion of TC-1 cells and allogeneic DC. The therapeutic efficacy was enhanced by co-administration of the vaccine with synthetic immunostimulatory ODN CpG 1826. PMID- 15783089 TI - Nucleoli in human early erythroblasts (K2, K1, K1/2 cells). AB - Human early erythroid precursors classified according to the nuclear size were studied to provide information on nucleoli in these cells using simple cytochemical procedures for demonstration of RNA and proteins of silver-stained nucleolar organizers. K2 cells with nuclear diameter larger than 13 microm and K1 cells with nuclear diameter larger than 9 microm corresponding to proerythroblasts and macroblasts (large basophilic erythroblasts) mostly possessed large irregularly shaped nucleoli with multiple fibrillar centres representing "active nucleoli". K1/2 cells with nuclear diameter smaller than 9 microm corresponding to small basophilic erythroblasts were usually characterized by the presence of micronucleoli representing "inactive nucleolar types". On the other hand, a few K1/2 cells contained large nucleoli with multiple fibrillar centres similar to those present in K2 cells and thus appeared as "microproerythroblasts". The nucleolar asynchrony expressed by the presence of large irregularly shaped nucleoli with multiple nucleoli (active nucleoli) and ring-shaped nucleoli (resting nucleoli) in one and the same nucleus of K2 or K1 cells was not exceptional and might reflect a larger resistance of these cells to negative factors influencing the erythropoiesis. The intranucleolar translocation of silver-stained nucleolus organized regions was noted in K2 cells and might indicate the premature aging of these cells without further differentiation. More studies, however, are required in this direction. PMID- 15783090 TI - Calcium absorption measurements in normal subjects in determining calcium deposition during prolonged hypokinesia and with and without calcium loading. AB - Measuring calcium (Ca) absorption, Ca balance and Ca level in serum,feces and urine during HK (hypokinesia) with and without Ca loading, the aim of this study was to disclose if prolonged HK could reduce Ca deposition more with or without Ca load contributing to greater Ca imbalance. Studies were conducted during 30 days pre-HK and 364-days HK. Forty male normal volunteers 23.7+/-6.0 years of age were chosen as subjects. They were divided into four groups: unloaded active control subjects (UACS), unloaded hypokinetic subjects (UHKS), loaded active control subjects (LACS), loaded hypokinetic subjects (LHKS). All hypokinetic subjects were walking average distances of 0.5+/-0.2 km day(-1), and active control subjects were running average distances of 6.6+/-1.2 km day(-1). LACS and LHKS were loaded with 1.3 mmol calcium lactate/kg body wt. Before Ca load, fecal Ca loss, urinary Ca and phosphate (P) losses, Ca imbalance, serum ionized calcium (CaI), P and total Ca (Ca(t)) levels increased significantly. (P < 0.05) with time, and serum intact parathyroid hormone (iPTH), 1.25 dihydroxyvitamin D (1.25(OH)2D3) levels and Ca absorption, decreased significantly (P < 0.05) with time in LHKS and UHKS compared with their pre-HK values and their respective active controls (LACS and UACS). After Ca load, however, Ca absorption, serum iPTH and 1.25 (OH)2D3 levels decreased significantly (P < 0.05) more with time, while fecal Ca loss, urinary Ca and P excretion and Ca imbalance increased significantly (P < 0.05) more with time in LHKS than UHKS. Conversely, before and after Ca load, fecal Ca excretion, urinary P and Ca loss, serum CaI, P, Ca, iPTH and 1.25 (OH)2D3 levels, Ca absorption and Ca balance did not change in LACS and UACS compared with their pre-HK values. The greater Ca losses with than without Ca load have shown that the more Ca is consumed the more Ca is eliminated during HK and Ca imbalance. The significant increase of Ca loss with Ca imbalance demonstrated reduced Ca deposition. Dissociation between Ca loss and Ca imbalance demonstrated reduced Ca deposition as the mechanism of Ca imbalance development during HK. PMID- 15783091 TI - Measurement of potassium absorption during hypokinesia in potassium supplemented and unsupplemented healthy subjects. AB - Measuring potassium (K+) absorption, and K+ levels in plasma, urine and feces during and after hypokinesia (HK) and K+ supplementation, the aim of this study was to determine if prolonged HK could depress K+ deposition significantly more with or without K+ supplementation. Studies were conducted during 30-days pre-HK, 364-days HK and 30-days post-HK. Forty male healthy volunteers 24.2+/-5.5 years of age were chosen as subjects. They were equally divided in four groups: unsupplemented active control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented active control subjects (SACS), and supplemented hypokinetic subjects (SHKS). Hypokinetic subjects were walking average distances of 0.5+/-0.2 km day(-1). Active control subjects were running average distances of 5.8+/-1.2 km day(-1). Both SHKS and SACS consumed daily 2.17 mEq elemental potassium per kg body weight. Potassium absorption, fecal and urinary K+ excretion, sodium (Na+) loss, plasma K+ and Na+ level and plasma aldosterone (PA) level did not change in SACS and UACS compared with their pre-HK values. During HK, K+ absorption decreased significantly (P < 0.05) with time, and fecal and urinary K+ loss, urinary Na+ loss, plasma K+ and Na+ levels and PA level increased significantly (P < 0.05) with time in SHKS and UHKS compared with their pre-HK values and their respective active controls (SACS and UACS). During initial 15-days of post-HK, K+ absorption increased significantly (P < 0.05), fecal and urinary K+ excretion, urinary Na+ excretion and plasma K+ and Na+ levels and PA level decreased significantly (P < 0.05) in hypokinetic compared with active control subjects; by the 30th day they approached the control levels. During HK and post-HK, K+ absorption, fecal and urinary K+ losses, urinary Na+ excretion, plasma K+ and Na+ levels and PA level, changed significantly (P < 0.05) more in SHKS than UHKS. Decreased K+ losses during post-HK showed K+ depletion during HK. Decreased K+ absorption with K+ depletion during HK showed decreased K+ deposition. The greater K+ changes in SHKS than UHKS, during HK and post-HK, demonstrated that K+ deposition decreased more with than without K+ supplementation. It was concluded that dissociation between K+ absorption and K+ depletion showed decreased K+ deposition as the main mechanism for K+ depletion during HK. PMID- 15783092 TI - Neonatal pyonephrosis--a case report. AB - Pyonephrosis is uncommon in adults as well as children and rarely reported in neonates. Candidial pyonephrosis in a neonate has been first reported in 1988 which was associated with iatrogenic perirenal collection [Cohen, HL, Haller, JO. J ultrasound Med 1988; 7(11): 647]. This is the second reported case of neonatal pyonephrosis and the first due to Staphylococcus aureus. PMID- 15783093 TI - Per urethra insertion of a through-and-through safety guide wire using a dual lumen ureteral catheter during percutaneous nephrolithotripsy. AB - BACKGROUND: The second ("safety") of two guide wires is commonly inserted antegradely in percutaneous nephrolithotripsy (PCNL). We describe the retrograde insertion of a through-and-through safety guide wire during PCNL. METHODS: After inserting a percutaneous nephrostomy 1 day earlier, a guide wire was introduced through the nephrostomy tube to the bladder and pulled out through the urethral meatus via a cystoscope and grasper. The tube was removed, leaving the wire positioned from the flank to the meatus. A dual-lumen catheter was introduced retrogradely through the urethra over the wire, up to the nephrostomy incision. The safety guide wire was introduced retrogradely through the catheter's other port. RESULTS: This procedure succeeded in 9 of 10 patients: the exception was very obese and the catheter was too short to reach the incision in the flank. CONCLUSION: Using a dual-lumen catheter allows quick and simple retrograde insertion of a safety guide wire during PCNL. PMID- 15783094 TI - Giant mixed-type perinephric liposarcoma. AB - Retroperitoneal fatty tumours are uncommon occurrences in current urologic practice, the differentials being lipomas, liposarcomas and extra-renal angiomyolipomas (AML). We report an unusual case of a 48-year-old Chinese female who presented with a 6.2 kg giant mixed-type liposarcoma in the left perinephric space, invading into the renal cortex and parenchyma. Extirpative surgery with left radical nephrectomy was performed, and histology confirmed liposarcoma with both myxoid and well-differentiated elements. The patient remains well and disease-free at 6-month follow-up. We review the literature on the diagnosis and management of these perinephric lipomatous lesions. We also discuss the diagnostic dilemma in differentiating such malignancies from benign angiomyolipomas in the perinephric area on conventional radiological imaging, and its direct implications with respect to extent of surgery and renal preservation. PMID- 15783095 TI - Tubercular mass mimicking a tumour in a horseshoe kidney: a unique presentation. AB - Tuberculosis of the kidney is quite a common disease and various forms of presentation are described. In most cases the disease results in atrophy, calcification or necrosis of parenchyma. The kidney is not generally palpable except occasionally in case of hydronephrosis; caused by upper ureteral structure. Renal tuberculosis presenting as large solid mass has not yet been reported. We present a case of solid renal mass of tubercular etiology. Diagnosis and management are presented. PMID- 15783096 TI - Renal cell carcinoma with giant common bile duct stone: a feasible surgical approach. AB - A 65-year-old man who presented with right renal mass and a giant CBD stone. The patient was managed with right radical nephrectomy and choledocotomy. Both the lesions of two different systems could be easily accessed via a subcostal incision in the same sitting. PMID- 15783097 TI - Renal cell carcinoma presenting as solitary foot metastasis. AB - Skeletal metastases from genitourinary tract are common, but metastatic tumors involving the hand and foot are rare. We herein present a case of 55-year-old man who presented with painful swelling of right foot and no urological complaints. Investigations revealed left renal mass and fine needle aspiration cytology from the swelling revealed findings consistent with metastatic clear cell carcinoma. PMID- 15783098 TI - Clitoral metastasis from transitional cell carcinoma of the renal pelvis: CT and MRI findings. AB - Metastatic tumors of the clitoris are extremely rare. We report a case of clitoral metastasis in a 75-year-old woman who was treated for transitional cell carcinoma of renal pelvis 2 years ago. The computed tomography and magnetic resonance imaging findings are presented with a short review of the literature. To the best of our knowledge, clitoral metastasis originating from transitional cell carcinoma of the renal pelvis has not been reported in the English language literature. PMID- 15783099 TI - Confirmation of the correct placement of lower end of DJ stent during open surgery: point of technique. AB - A simple technique to confirm the correct placement of the double J (DJ) stent at the time of antegrade insertion during open surgery is described. At the time of antegrade DJ stent placement about 30-50 ml of sterile methylene blue is instilled into the bladder. Appearance of methylene blue from the upper end and side holes of the DJ stent confirms the correct placement of the lower end of the stent. PMID- 15783100 TI - Transitional cell carcinoma of ureteral stump after radical nephrectomy in a patient with a history of bladder carcinoma. AB - Transitional cell carcinoma (TCC) of ureteral stump after radical nephrectomy is rare. Following nephrectomy patients with a prior history of bladder cancer must have their ureteral stumps evaluated. Furthermore, the presence of hematuria should alert the urologist to a possible TCC in the ureteral stump. We present a patient who developed TCC of the ureteral stump after radical nephrectomy. PMID- 15783101 TI - Anurea caused by primary amyloidosis of the lower third of the ureters, the ureterovesical junction and the urinary bladder: a case report and review of the literature. AB - Amyloidosis is a systemic disease affecting many organs, characterized by the deposition and accumulation of hyaline, eosiniphilic and proteinaceous material in various tissues. This report presents a patient with anurea caused by primary amyloidosis of the lower third of the ureters, the ureterovesical junction and the urinary bladder. PMID- 15783102 TI - Complication of percutaneous suprapubic cystostomy. AB - A patient with urethral stricture presented to us with acute retention. Sequential dilatation over a guidewire was done and a suprapubic catheter was placed percutaneously. Guide wire broke during the procedure, which came to our notice on micturating urethrogram. Laser core through for the stricture and endoscopic removal of the guidewire were done. PMID- 15783103 TI - Intravesical stone formation on intrauterine contraceptive device. AB - Since more than 30 years, intrauterine contraceptive devices (IUCD) have been used for a contraceptive opportunity. Although they are termed to be a safe and effective method for contraception, they also have some type of complications and uterine perforation, septic abortion, pelvic abscess are the serious complications of these devices. The incidence of uterine perforation is very low, but in the literature nearly 100 cases were reported about the extra uterine localization of IUCD. Migration may occur to the adjacent organs. We here in describe a case of a 31 year-old woman who had an IUCD with stone formation in the bladder. In the literature all of the cases were reported as IUCD migration, but although it seems technically impossible, IUCD placement into the bladder should also be considered in misplaced IUCDs. PMID- 15783104 TI - A case of severe hemorrhagic cystitis following bone marrow transplantation. AB - Hemorrhagic cystitis (HC) is a common adverse effect caused by the preparative regimens for bone marrow transplantation. Unfortunately, an effective therapy for HC has not yet been established. Here we report a case of severe HC in which the upper as well as the lower urinary tract were severely affected, resulting in right ureteral obstruction and a contracted bladder. Hyperbaric oxygen therapy was effective in suppressing the severe HC, but the cause of HC could not be identified. Five years after the bone marrow transplantation the patient underwent augmentation ileocystoplasty with ileal ureteral replacement. PMID- 15783105 TI - Radical cystectomy in septuagenarian patients with bladder cancer. AB - INTRODUCTION: Clinical results of radical cystectomy performed on elderly bladder cancer patients over 70 years old were assessed to determine whether age is one of the critical points for the application of this type of surgery. MATERIALS: From January 1992 to December 2002, 41 radical cystectomy performed in septuagenarian population were compared with 197 performed in patients less than 70 years of age. RESULTS: The early and late complication rate for septuagenarians was 29.7% and 12%, compared with 26.9% and 9.6% for patients younger than 70 years respectively. The overall mortality rate for septuagenarians was 4.9%, compared with 8.6% for patients younger than 70 years. There was no significant difference between the two groups with regard to pathologic stage or length of hospital stay. The 5-year overall survival rate for septuagenarians was 53% compared with 59% for patients younger than 70 years. CONCLUSIONS: When indicated after adequate preoperative assessment and optimization of the patient, radical cystectomy is a safe procedure in the septuagenarians and patient should not be denied surgery dependent on chronologic age. PMID- 15783106 TI - Pilot attempt of advanced prostate cancer treatment T3NxMx-1 by intermittent more complete androgen blockade. AB - The aim of the prospective pilot study was evaluating efficacy and tolerance of pharma-cological more complete androgen blockade (mMAB) by using Zoladex LA 10.8 mg, Casodex 50 mg and Proscar 5 mg in patients with advanced prostate cancer (T3,Nx Mx-1). METHODS: This five-year study involved 14 patients aged 67-82 years (average 73). Zoladex LA was administered subcutaneously every 3rd month of treatment, and every day 1 tablet Casodex and 1 tablet Proscar. In the time when PSA was <0.1 mg/ml Zoladex and Casodex were withdrawn, and only Proscar was left. The mMAB treatment was resumed when PSA > 0.1 ng/ml. Before and every 3 months the following laboratory tests were made: PSA, sedimentation, bilirubine, transaminase, phosphatase, ultrasonography (USG); and adverse events were registered. The following criteria of assessment were adopted: CR--complete response--examination tests normal, improved condition, reduction of prostate dimension in USG and value of PSA < 0.1 ng/ml; PR partial response i.e. no progression, the PSA level drops down to the reference values and a reduction of prostate dimension in USG occurs, NR--no response i.e. progression, increased prostate dimension in USG and/or metastases in scintygraphy, as well as PSA above normal. RESULTS: The mean followup time was 60 months. After the initial 6 months two patients were off-therapy mMAB because they were qualified for radiotherapy. A successive patient (no. 5) was off-therapy (after 4 years) because he left Warsaw to go abroad. After 60 months results mMAB were based on 11 patients' records, and a complete response was confirmed in 7 patients, partial response in 3 patients and no response in one patient. CONCLUSION: Results of this study show that pharmacological intermittent mMAB is an efficient way of treating advanced prostate cancer. Side effects are low and occur in moderate intensity and do not oblige to treatment withdrawal. PMID- 15783107 TI - Prostate cancer with bladder neck involvement: pathologic findings with application of a new practical method for tumor extent evaluation and recurrence free survival after radical prostatectomy. AB - BACKGROUND: Recent studies have questioned the high risk for disease recurrence in cases of bladder neck involvement by prostate cancer (pT4 disease). DESIGN: The study was based on 141 patients submitted to radical prostatectomy. PSA recurrence was defined as 0.5 ng/ml or rising at three different examinations. Bladder neck invasion was correlated to Gleason score (<7 or > or =7), urethral or lateral positive surgical margins, tumor size (extensive or not extensive) using a new simple point-count method, race and preoperative PSA levels (<10 or > or =10 ng/ml). The mean and median follow-up periods were 17.2 and 15 months (range 3-55). Recurrence-free survival curves were constructed using the Kaplan Meier survival method with tests of significant differences based on the log-rank statistic. RESULTS: Bladder neck invasion was seen in 30/141 (21.27%) patients; 9 (6.38%) had also concomitant positive surgical margins in the sections. Extraprostatic extension was seen in 39/141 (27.65%) and seminal vesicle invasion in 18/141 (12.76%) patients. Patients with bladder neck invasion significantly correlated to Gleason score (P = 0.04), preoperative PSA (P < 0.001), and tumor extension (P= 0.04). No correlation was found to age (P = 0.44), race (P = 0.12) and positive urethral or lateral margins (P = 0.32). The PSA-recurrence relative risk in patients with bladder neck invasion was 0.17 (P = 0.68), with extraprostatic extension 0.53 (P = 0.47) and with seminal vesicle invasion 5.76 (P = 0.02). CONCLUSIONS: Bladder neck involvement correlates with pathologic unfavorable findings on radical prostatectomy specimens as well as to preoperative PSA levels. However, the PSA-recurrence risk associated with bladder neck involvement (pT4) was similar to extraprostatic extension (pT3a) and substantially lower than seminal vesicle invasion (pT3b). The findings favor a need for downstaging of bladder neck involvement in the next version of the TNM system. The method proposed for tumor extent evaluation is simple and accessible to all pathologists working in routine pathology laboratories. PMID- 15783108 TI - Important preoperative prognostic factors for extracapsular extension, seminal vesicle invasion and lymph node involvement in cases with radical retropubic prostatectomy. AB - INTRODUCTION: Initial diagnostic evaluation may provide information about the extent of disease after radical retropubic prostatectomy (RRP). The aim of this study was to investigate the predictive value of preoperative serum prostate specific antigen (PSA) level, local disease extension identified by transrectal ultrasound (TRUS), total number of positive biopsies and percentage of positive cores for cancer, as well as TRUS Biopsy Gleason score in determining the extent of disease in radical retropubic prostatectomy specimens. MATERIALS AND METHODS: A retrospective analysis was performed on 171 patients who underwent RRP from March 1993 to February 2003 for organ confined prostate cancer and whose follow up data was accessible. The correlation of preoperative serum PSA level, local disease extension in TRUS, the total number of positive sextant biopsies and the percent of cores positive for cancer and Gleason score at TRUS biopsy specimen with the extent of disease at final pathology (Extra-capsular extension (ECE), seminal vesicle invasion (SVI), lymph node involvement (LNI) and surgical margin (SM) status on RRP specimens) were analyzed. RESULTS: The median age of the patients was 65 years. The mean preoperative serum PSA level of all patients was 11.6+/-1.2 (median 8.6) ng/ml. Histopathological evaluation of RRP specimens revealed 60 (35%) patients with ECE, 38 (22.2%) with SVI, 7 (0.04%) with LNI, and 58 (33.9%) had positive SM. Comparing the preoperative TRUS findings and postoperative evaluation of RRP specimens, the sensitivity of TRUS in predicting the ECE was 11.8% and specificity was 96%. Sensitivity of TRUS in predicting SVI was 9.8% and its specificity was 99%. With univariate analysis (sample t-test), Gleason score, percent of cores positive for cancer, and DRE were found to be predictive factors for extra-prostatic disease in RRP specimens. But with multivariate analysis (logistic regression test) Gleason score appears to be the most important and independent predictive factor for extra-prostatic disease in RRP specimens. Serum PSA levels and percentages of cores positive for cancer were also significant predictors of non organ-confined disease found at final pathology. CONCLUSION: Gleason score is the most important and independent predictive factor for extra-prostatic disease. Serum PSA levels and percentages of cores positive for cancer are the other important but non-independent predictive factors. PMID- 15783109 TI - Bulbar urethral polyp mimicking a urethral stricture--report of case with review of literature. AB - Benign anterior urethral fibroepithelial polyps in the adult are extremely rare lesions. We report the presentation and management of such a case of recurrent bulbar urethral poly in a young adult who had presented with obstructive voiding symptoms mimicking a urethral stricture. He was successfully managed by urethrotomy excision and primary urethral repair. PMID- 15783110 TI - Penile amputation and emasculation: hazards of modern agricultural machinery. AB - We report a case of 20-year-old farmer with penile amputation and emasculation secondary to crushing injury inflicted by agricultural machinery. His entire penis, scrotum and both the testis had been avulsed after falling onto a rotating axel wheel of an electronically driven thrashing machine. After resuscitation the wound was thoroughly washed with saline, hemostasis achieved and a perineal urethrostomy was made from the remnant urethra. PMID- 15783112 TI - Seminoma--a unique case of tumor located in the head of epididymis. AB - We report a very rare case of tumor in the head of left epididymis without localized primary foci of tumor in the testis. PMID- 15783111 TI - Is dorsale penile vein ligation (dpvl) still a treatment option in veno-occlusive dysfunction? AB - PURPOSE: To determine the long-term results of the DPVL for the treatment of venous impotence. PATIENTS AND METHODS: The long term results of DPVL in 134 patients were interviewed. The mean age was 39.2 (range 21-72). Power color doppler imaging, pharmacocavernography/dynamic cavernosometry were performed. Postoperatively, all the patients were controlled in the 6th month, first year and, if possible, once a year. The mean follow-up was 54.8 (14-76) months. Postoperative outcomes were classified into three groups: complete spontaneous erection (CR), response to pharmacotherapy (PR) or no satisfactory improvement (NR). RESULTS: The short-term success in the 6th month according to above mentioned was 38.8%, 18.6% and 42.5%; and in the first year was 19.4%, 14.9% and 65.6% respectively. The result in 35 patients whose follow-up was at least 5 years (mean 67 months) was 11.4%, 14.3% and 74.3% respectively. Positive prognostic factors were preoperative age <40, duration of erectile dysfunction <2 years, non-smoker patients, non neurogenic disease and distal disease. With all these parameters present, long-term success (CR, PR) rose from 33.6% to 55.9% (P < 0.001). CONCLUSIONS: Long-term success for unselected patients undergoing DPVL is disappointing; however, careful patient selection significantly improves long term results. PMID- 15783113 TI - Primary anaplastic large cell lymphoma of the testis. AB - Anaplastic large cell lymphoma (ALCL) was first described by Stein et al. in 1985, at that time neoplastic cells were labeled by the monoclonal antibody CD30. ALCL was included as a differentiate entity in the reviewed Kiel and REAL classification. ALCL carries the t (2; 5) (p23; q35) translocation; the absence of ALK kinase from normal lymphoid cells indicates that immunohistochemical expression of ALK is specific for the (2; 5) translocation. This disease is characterized by a diffuse proliferation of large anaplastic cells with kidney shaped/horse-shoe nuclei. A distinguishing feature is a perinuclear eosinophilic region that represents a prominent Golgi apparatus. These cells are named hallmark-cells being almost pathognomonic. Immunohistochemically the most important features are that tumor cells consistently express CD30 and EMA on the cell membrane and in the Golgi region, while ALK immunostaining is usually both, cytoplasmic and nuclear. To our knowledge only two cases of primary ALCL of the testis have been reported. Hereby we present a case of a typical ALCL expressing ALK and CD30, which presented with subcutaneous nodules and bilateral testicular mass, without systemic involvement. PMID- 15783114 TI - A life-threatening hematochesia after transrectal ultrasound-guided prostate needle biopsy in a prostate cancer case presenting with lymphedema. AB - A 62-year-old male presenting with lymphedema at the left lower extremity was diagnosed as prostatic carcinoma by an ultrasound-guided needle biopsy. Six days after the biopsy procedure, he developed severe hematochesia. Colonoscopy revealed an oozing visible vessel at the anterior rectal wall, a few centimeters from the anus. Endoscopic hemostasis with %1 athexysclerol was applicated with success. Rebleeding with same severity as the first one occurred 48 hours after the first bleeding and endoscopic hemostasis was successful again. Follow-up was uneventful. PMID- 15783115 TI - Microvessel density as a prognostic marker in bladder carcinoma: correlation with tumor grade, stage and prognosis. AB - INTRODUCTION: The aim of our study was to evaluate tumor angiogenesis as a prognostic marker of transitional cell carcinoma of the bladder and to asses its relationship to established variables for survival and response to therapy. PATIENTS AND METHOD: Microvessel density (MVD), a measure of tumor angiogenesis, were evaluated in 77 primary bladder cancers. Forty-three superficial carcinomas and 34 invasive carcinomas were analysed. Tumor specimens of all patients were obtained by transurethral resection (TUR) and all the tumors were transitional cell carcinomas. Twenty-two patients with invasive bladder cancer have undergone M-VEC chemotheraphy. The correlation between MVD and histopathological grade, tumor stage and prognosis was evaluated. MVD was identified by immunostaining of endothelial cells using anti-CD34 antibody. For statistical analysis Kruskal Vallis, Mann-Whitney U and Fisher's exact tests were used. RESULTS: MVD was correlated with tumor grade, stage and prognosis. Significantly higher MVD was determined in invasive tumors than superficial tumors (p < 0.05). MVD increased with tumor grade and stage (p < 0.05). High MVD was correlated with the risk of clinical progression in both superficial and invasive bladder carcinomas (p < 0.05, p < 0.001 respectively). Invasive tumors with remission after M-VEC chemotheraphy had lower MVD than tumors with progression after M-VEC. CONCLUSION: These data demonstrate that MVD in bladder carcinoma correlates with grade, stage and malignant potential of the tumor. Quantification of tumor angiogenesis may allow selection of the type of treatment for bladder cancer patients. PMID- 15783116 TI - Abdominal polyorchidism: a case report and review of the literature. AB - Abdominal polyorchidism is rare. We report a case and review the literature. We suggest a very watchful abdominal evaluation of patients for a second ipsilateral testis who are operated for undescent testis. Polyorchidism is a very rare anomaly which requires definitive histological diagnosis. Although imaging techniques may be helpful in the diagnosis, it's mostly incidentally found in surgical exploration. Patients may present with pain or scrotal swelling. Most commonly abnormalities are maldescent (40%), hernia (30%), torsion (15%), hydrocele (9%) and malignancy (6%). PMID- 15783117 TI - Intraparenchymal renal artery aneurysms. Case report with review and update of the literature. AB - Increased interest in aneurysms involving the renal artery and its branches has occurred during the past 3 decades. The prevalence of renal artery aneurysms is approximately 0.01%-1% in the general population as well as 2.5% in hypertensive patients undergoing angiography. Intraparenchymal renal artery aneurysms (IPRAAs) are rare since being detected in less than 10% of patients with renal artery aneurysms. The Authors report an unusual case of multiple small intrarenal artery aneurysms associated with a large IPRAA located in the mid portion of the right kidney. Usually, IPRAAs are secondary to diseases or injuries of the kidney vascular network. They are classified as true, false, saccular, fusiform, dissecting, and microaneurysms. Potential complications of IPRAAs include peripheral dissection, thrombosis, hypertension, renal infarction and rupture. IRAAs may be detected incidentally as well as present with urologic symptoms and signs related to complications. Actually, IRAAs are investigated by non invasive modalities including duplex ultrasound, magnetic resonance angiography, spiral three-dimensional computed tomography angiography, and three-dimensional reconstructed rotational digital substraction angiography of the segmental and distant branches of the renal artery. Angiography with intrarterial injection of contrast material is the gold standard in diagnosing IPRAAs. Treatment options for IPRAAs include observation, aneurysmectomy with surgical repair, endovascular procedures, nephrectomy or partial nephrectomy. Observation is indicated for asymptomatic intraparenchymal renal artery aneurysms measuring less than 2 cm in diameter. Surgical repair of IPRAAs includes aneurysmectomy and reconstruction of the renal artery by in vivo or ex vivo technique. The procedure is indicated for IPRAAs causing renovascular hypertension, dissection, urologic symptoms, embolization, local expansion and women of childbearing age with a potential for pregnancy. In recent years, transcatheter arterial embolization has emerged as a simple, useful and effective technique in managing IRAAs. The procedure is performed by transfemoral catheterization as well as by superselective catheterization and embolization of interlobar arteries with 3F microcatheters. Endovascular occlusion is obtained by using gelatin sponge, steel coils, detachable baloons, and conventional non-detachable microcoils delivered through a microcatheter. Nephrectomy or partial nephrectomy are reserved for conditions precluding renal revascularization which include overt RAA rupture, covert RAA rupture, artery-to-vein fistula, renal cell carcinoma, end stage nephropaty, renal infarction, severe ischemic renal atrophy or complex intrarenal aneurysms. Recently, partial nephrectomy by the laparoscopic approach has been proposed for managing IPRAAs and the procedure is considered feasible and safe. PMID- 15783118 TI - Immunohistochemical expressions of uPA and its receptor uPAR and their prognostic significant in urinary bladder carcinoma. AB - BACKGROUND: The authors found previously that plasma levels of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) were elevated in patients with bladder carcinoma and were associated with features of biologically aggressive disease. The present study has been performed to analyze the expressions of two antigens by immunohistochemical staining in bladder transitional cell carcinoma. PATIENTS AND METHODS: The tumors from 72 men and 28 women with a mean age 46.15 years (range 30-67 years) were examined. Paraffin sections of 5 microm thickness were prepared for immunohistochemical staining of uPA and uPAR antigens. Age, sex, tumor grade and stage, DNA ploidy, lymph node status, and metastases were evaluated in relation to outcome. Univariate and multivariate analysis of survival were performed. RESULTS: The overall 5-year survival was 66%. Thirty six and 46 cases were positive for uPA and uPAR expressions, respectively. In univariate analysis, tumor stage, lymph node status, metastases, uPA and uPAR have a significant impact on the survival for these patients. In a Cox proportional hazard model, uPAR sustained its significant impact on survival. CONCLUSIONS: These findings suggest that uPAR is an independent additional prognostic factor in patients with transitional cell carcinoma of the bladder. PMID- 15783119 TI - Prospective randomised trial of isoniazid prophylaxis in renal transplant recipient. AB - Renal transplantation (RT) recipients are at a high risk of developing tuberculosis (TB) following transplantation. Effectiveness of isoniazid (INH) in preventing TB is well documented in immunocompetent as well as immunocompromised persons. There is paucity of data on role of INH prophylaxis in RT recipients. Thus, a prospective randomised trial of INH in RT recipients was carried out to determine the efficacy of daily INH monotherapy in the prevention of TB in these patients. Patients of end stage renal disease (ESRD) taken for RT formed the subjects of study. Patients with active TB and active hepatitis at the time of RT were excluded from the study. Patients were randomised to receive INH 300 mg with pyridoxine 20 mg daily from the day of RT. The duration of the treatment was planned for 1 year or till the development of TB, which ever was earlier. Between October 1998 and September 2000, 114 RT were done at our hospital. Of these, 24 (21%) patients had active TB at the time of RT and thus were excluded. Patients included were randomised with 1:2 ratio of treatment and control group. Of the 90 patients thus enrolled, 30 were randomised in treatment group and 60 in control group. Of the included patients five patients had very early graft loss (three in treatment and two in control group) within days and thus excluded from the analysis. Three of the 27 (11.1%) patients in treatment group and 15 (25.8%) in control group developed TB (P = 0.10). The risk ratio of (RR) of INH versus control group of TB was 0.36 (95% CI, 0.10-1.32) but the difference was not statistically significant (P = 0.12). Only one patient developed INH induced hepatitis. In conclusion, with INH prophylaxis, there was a trend towards protection from TB, though it was not statistically significant. Further, all patients tolerated INH and hepatotoxicity was not a major problem in this group of patients. PMID- 15783120 TI - Fibrosing cholestatic hepatitis in renal transplant recipient with CMV infection: a case report. AB - Fibrosing cholestatic hepatitis (FCH) is an uncommon complication of renal transplantation. It is usually associated with hepatitis B and C viral infection. It is further rare in renal transplantation in absence of HBV and HCV infection. To the best of our knowledge, only three cases of FCH in renal transplantation, which were both HBV and HCV negative, have been reported to date. Out of these, two cases were diagnosed to have CMV infection and the third was attributed to azathioprin. We are presenting another case of FCH in a renal transplant recipient with CMV infection. PMID- 15783121 TI - Improvement in uremic symptoms after increasing daily dialysate volume in patients on chronic peritoneal dialysis with declining renal function. AB - OBJECTIVE: Patients on peritoneal dialysis (PD) can develop uremic symptoms as their residual renal function declines. In this retrospective study, we assessed the effect of increasing the dose of dialysis in patients who developed uremic symptoms. METHODS: Patients on PD who had an increase in their dialysis dose due to either the appearance of uremic symptoms or to worsening biochemical parameters were included in this study. These patients had to have been on PD for at least 6 months before and after the increase in their dialysis dose. Patients whose dialysis dose was increased after the initial Adequest (done within 2-3 months of starting PD) findings or for reasons other than underdialysis were excluded from this study. The symptoms studied in 104 patients included fatigue, anorexia, insomnia, pruritus and nausea. The presence or absence of theses symptoms was evaluated before and after the increase in the dialysis volume. Several clinical and laboratory data including the adequacy results were compared before and after the increase in dialysis dose. RESULTS: Patients were on PD for 24.6+/-16 months when dialysis dose was increased. Eighty-five (82%) of them were on continuous ambulatory peritoneal dialysis (CAPD) while the remaining were on continuous cycler peritoneal dialysis (CCPD). Fatigue was the most common symptom that led to an increase in the dialysis dose (64%). The prevalence of all the symptoms studied decreased significantly after the increase in dose of dialysis. The weekly peritoneal creatinine clearance increased from 47.35+/-0.88 to 57.34+/ 1.401 (P < 0.0001) and the weekly Kt/V increased from 1.8+/-0.03 to 2.27+/-0.05 (P < 0.0001). The daily urine volume and the residual GFR decreased from 318.7+/ 36.4 to 151.9+/-22.8 ml/day (P < 0.0001) and 2.05+/-0.2 to 0.82+/-0.12 ml/min (P < 0.0001) respectively during the study period. CONCLUSION: The prevalence of all uremic symptoms decreased significantly after the daily dialysate volume was increased. The improvement in symptoms despite the decline in residual function emphasizes the beneficial effect of increased dialysate volume, which produced a significantly higher peritoneal creatinine clearance and Kt/V after the change in the PD prescription. PMID- 15783122 TI - Gestational urinary hyperthiosulfaturia protects hypercalciuric normal pregnant women from nephrolithiasis. AB - Urinary calcium excretion increases by 1-2-fold during gestation in normal, uncomplicated pregnant women. Hypercalciuria occurs in all trimesters and elevates urine supersaturation with regards to calcium oxalate. However, crystalluria has not been a frequent clinical finding and stone formation is not a common complication of pregnancy. To elucidate this discrepancy we measured various chemical entities (i.e. calcium, oxalate, uric acid, phosphorous, magnesium, citrate, sulfate and thiosulfate) in urine at the end of each trimester of 25 pregnant women. Twenty-five healthy women served as controls. Our observations show that endogenous thiosulfate, a natural component of urine, increased considerably during pregnancy to approximately 36, 38 and 40 microM/24 hour at the end of each three trimesters. One month after delivery, endogenous thiosulfaturia and hypercalciuria, in parallel, returned to initial normal values. Consequently, it seems that gestational hyperthiosulfaturia protects hypercalciuric normal pregnant women from the risk of nephrolithiasis. PMID- 15783123 TI - Renal transplant recipients over aged 60 have diminished immune activity and a low risk of rejection. AB - Strict consideration of the renal transplant candidate's chronologic age is generally supplanted by more subjective reflection on his (her) physiologic state. In the US, patients over 64 years old represented 9.0% of renal transplant recipients in the year 2000, yet little prior experience is available with which to guide the management of geriatric patients. Two hundred and forty six consecutive recipients of primary kidney transplants at the Yale-New Haven Organ Transplant Center between 1990 and 1995 were included in an outcome analysis. Age at transplantation ranged from 2 to 68 years; the study group consisted of the 16 (6.5%) over age 60. The immunosuppressive protocol was uniform for all patients. There was a disproportionately high use of cadaveric organs by older patients; only 1/16 (6.3%) received a living donor kidney. The overall rate of rejection within the first 90 days was 6.7% of cadaveric recipients over 60 versus 37.6% of younger recipients, P=0.001. Actual patient survival rates at 6 years were 100% of patients younger than 11 years versus 69% (11/16) of those older than 60 years. Death censored 5 year graft survival was 100% in older patients versus 85% among the younger patients. The older and younger patients received quantitatively equivalent immunosuppression, but acute rejection was uncommon in the former (6%) versus the younger cohort (34%). It seems logical to consider whether older renal transplant recipients may benefit from a less aggressive immunosuppression strategy. PMID- 15783126 TI - Potassium metabolism in patients with chronic kidney disease. Part II: patients on dialysis (stage 5). AB - Potassium is removed mainly by diffusion during dialysis. In hemodialysis, potassium removal averages 70-150 mmol per session, and the presence of glucose free dialysate, sodium profiling, and hyperkalemia, may increase its removal. The most frequent potassium derangement in hemodialysis patients is hyperkalemia. Hemofiltration removes approximately 60 mmol of potassium per treatment. In peritoneal dialysis patients, despite lower potassium removal (about 30-40 mmol/day), hypokalemia is the most frequent electrolyte alteration, probably due to movement of potassium into the cells mediated by insulin, secondary to glucose absorption from the dialysis solution. PMID- 15783125 TI - Potassium metabolism in patients with chronic kidney disease (CKD), Part I: patients not on dialysis (stages 3-4). AB - Potassium is the main cation in the intracellular space and its content in healthy adults is around 3700 mmol. Its retention is one of the dangers that accompanies chronic renal failure, especially when glomerular filtration rate (GFR) falls toward 20% of normal. There exist different mechanisms by which hyperkalemia is avoided in patients with renal failure. These include spontaneous restriction of dietary potassium intake to 40-60 mmol/day and aldosterone induced increase in potassium excretion in renal distal tubules and colonic epithelium. Although renal and intestinal adaptation are important in maintaining overall potassium balance during CKD, movement of potassium into the cells is extremely important in the body's acute defense against hyperkalemia, and occurs daily during postprandial potassium overload, through shift of this cation into hepatic cells mediated by insulin and thus avoiding post-prandial hyperkalemia. The above described mechanisms contribute to maintenance of normal serum potassium in patients with CKD. PMID- 15783124 TI - Urinary tract infections in the frail elderly: issues for diagnosis, treatment and prevention. AB - Urinary tract infections (UTIs) are among the most common infections in frail elderly adults, whether they are community dwelling, live in long term care facilities (LTCFs) or are hospitalized. UTIs cause substantial morbidity and mortality in frail elderly men and women. While many major risk factors in these individuals may not be modifiable, improved attention to incontinence management, judicious use of antimicrobials and urinary catheters, and, in women, appropriate use of topical estrogen may be useful in reducing UTIs. Future strategies may also include the appropriate use of new urinary catheter technology and emerging vaccines. PMID- 15783128 TI - OIG 2005 work plan outlines areas for compliance review. PMID- 15783127 TI - Clinical physiology and pharmacology conference: a nonpharmacological, evidence based medical approach using exercise method to lower and maintain blood pressure control in the elderly. PMID- 15783129 TI - Surveying the prevalence of asthma, allergic rhinitis and eczema in school children in Khon Kaen, Northeastern Thailand using the ISAAC questionnaire: phase III. AB - This is the second survey of schoolchildren in Khon Kaen, Northeastern Thailand, using the Thai version of the ISAAC questionnaire to examine the trend in the prevalence of asthma, allergic rhinitis and eczema, and to compare the results with the ISAAC Phase I data. We analyzed 5,075 questionnaires comprising 2,119 six- to seven- and 2,956 thirteen- to fourteen-year-old children (48 and 42 percent male, respectively). The cumulative vs. 12-month prevalence according to the written questionnaires were: 14.3 vs. 9.8% for wheezing, 42.6 vs. 33.3% for rhinitis and 13.5 vs. 11.2% for eczema, respectively. The cumulative vs. 12-month prevalence for the wheezing module, based on the video questionnaire, was 9.2 vs. 6.3%, respectively. Most Phase III prevalence was significantly lower than the first survey except for the steady, 12-month prevalence of wheeze. Our study confirms the high prevalence of allergic diseases among school-children in Northeastern Thailand; albeit, prevalence has not increased in recent years. The Thai version of the English-language ISAAC questionnaire needs to be validated before further use in epidemiological research. PMID- 15783130 TI - Sensitization to common allergens, especially pollens, among children with respiratory allergy in the Trakya region of Turkey. AB - Asthma and allergic rhinitis are common problems in children and the causative pollen allergens vary according to the geographical area. The aim of this study was to investigate patterns of sensitization to common inhalant allergens, especially pollens, in Turkish children living in the Trakya region and to determine differences between rural and urban areas. Allergen skin testing was prospectively performed on 539 children aged between 4 and 17 years with respiratory allergy. The reaction was considered to be positive if the mean wheal diameter was at least 3 mm greater than that of the negative controls. We detected positive skin reactions in 420 (77.9%) children. Two hundred and eighty one (52.1%) mite, 277 (51.4%) pollen, 174 (32.3%) mold, 65 (12.1%) animal dander, 12 (2.2%) cockroach and 6 (1.1%) latex skin sensitivities were detected. Among the pollen allergies 173 were cereal pollen (32.1%), 170 grass pollen (31.5%) and 144 tree pollen allergies (26.7%). The most common positive skin test among the pollens was to cultivated wheat (Titicum vulgare) (n = 116, 21,5%), followed by rye grass (Lolium perenne) and orchard grass (Dactylis glomerata). Positive skin reactions to Alternaria, to Candida albicans, and to all pollens except Ulmus competris, Pinus sylvetris, Platanus vulgaris and Tilia platyphyllos, were higher in children with allergic rhinitis than in those with asthma. In children from rural areas, allergic skin reactivity was found to be more common against Candida albicans, sheep dander and all pollens except Corylus avellana, Fraxinus excelsior, Populus alba, Pinus sylvetris, Platanus vulgaris and Chenopodium album, than in urban children. Although Trakya is close to Greece and other Mediterranean countries, this study suggests that the pollens, which sensitize children, are not similar. PMID- 15783131 TI - The prevalence of exercise-induced bronchoconstriction among symptomatic sportsmen. AB - Exercise-induced bronchoconstriction (EIB) is a transient airflow obstruction that usually occurs 5 to 15 minutes after physical exertion. Although this condition is preventable, it is still underrecognized and affects the quality of life. The purpose of this study was to evaluate the prevalence of EIB, to find out whether self-reported symptoms were enough to establish the diagnosis of EIB and to define related symptoms and risk factors among symptomatic students of a Sports Academy in our city, which is situated at about 2,000 meters altitude and experiences a long winter period. A questionnaire was sent to students of the Sports Academy. After its evaluation, symptomatic students were invited to perform a pulmonary function test (PFT) before and after a six minute exercise test. Two hundred and seventy-seven students (205 males and 72 females, mean age: 20+/-4) from 12 different types of sport replied to the questionnaire. Of these subjects, 43% were symptomatic. Out of the 119 symptomatic students, 63 accepted our invitation. Three of them were excluded. After performing a six-minute exercise test, EIB was observed in 5 (8%) of them, whose symptoms were aggravated by cold weather. Our results suggest that EIB may be common among symptomatic sportsmen. Screening is important, since EIB could be prevented with appropriate diagnosis and treatment. Although respiratory symptoms during exercise may give important clues, only these symptoms are not sufficient to establish the diagnosis of EIB. Symptomatic patients should undergo further tests for EIB. PMID- 15783132 TI - Is the menstrual cycle affecting the skin prick test reactivity? AB - Allergen skin prick tests (SPT) are very sensitive and specific tests to detect allergic sensitization in atopic patients. Certain factors like antihistamines, antidepressant therapies or circadian rhythms can alter the results of SPT. In women, the changes in endogenous hormone levels throughout the menstrual cycle may affect the allergic responses and natural course of allergic diseases. The aim of this study was to investigate the probable influence of the phases of the menstrual cycle on SPT reactivity to allergen extracts and histamine. Forty-two female patients with seasonal allergic rhinoconjunctivitis were enrolled in the study. Skin prick test reactivities to allergens and histamine were measured at the beginning of the menstrual cycle (3rd or 4th day), mid-cycle (14th or 15th day) and end-cycle (27th or 28th day) consecutively. Serum estradiol, progesterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) levels were determined simultaneously. We observed the most significant reactions to allergens when SPT is performed at mid-cycle. However, SPT reactivity to histamine did not vary throughout the menstrual cycle. Serum estradiol and LH levels showed positive correlation with SPT reactivity to allergens at mid-cycle. Our results suggest that SPT give the best results when they are performed at mid cycle. Additionally, allergens seem to cause mast cell degranulation to a greater extent in subjects in which endogenous hormones like estradiol and LH are elevated. PMID- 15783133 TI - Modulation of tryptase and histamine release from human lung mast cells by protease inhibitors. AB - Inhibition of IgE dependent histamine release from human mast cells by protease inhibitors has been observed in skin, tonsil and synovial tissues. However, little is known about the actions of protease inhibitors on tryptase release from human lung mast cells. We therefore examined the ability of protease inhibitors to modulate tryptase and histamine release from human lung mast cells. IgE dependent tryptase release from dispersed lung mast cells was inhibited to a maximum of approximately 53.8% and 44.5% by N-a-tosyl-L-lysine chloromethyl ketone (TLCK) and N-p-Tosyl-L-phenylalanine chloromethyl ketone (TPCK), respectively. A similar degree of inhibition of calcium ionophore A23187 (CI) induced tryptase release was also observed with these two inhibitors. Preincubation of TLCK or TPCK with the mast cells at 37 degrees C for 20 minutes before addition of anti-IgE or CI did not improve their ability to inhibit anti IgE and CI induced tryptase release. At a concentration of 10 microg/ml, protamine inhibited anti-IgE or CI induced tryptase release; but at 100 microg/ml, it increased anti-IgE and CI induced release of tryptase from lung mast cells. A concentration dependent inhibition of anti-IgE and CI induced release of histamine from lung mast cells was also observed with TLCK, TPCK and protamine. The maximum inhibition of anti-IgE induced histamine release was approximately 40.7%, 40.2% and 33.4% with TLCK, TPCK and protamine, respectively. At the concentrations tested, TLCK and TPCK by themselves did not stimulate tryptase and histamine release from lung mast cells. A specific inhibitor of aminopeptidase, amastatin, had no effect on anti-IgE induced tryptase and histamine release and was used as control. In conclusion, it was demonstrated that protease inhibitors are able to inhibit IgE dependent tryptase and histamine release from human lung mast cells, which suggested that they could be developed to a novel class of anti-inflammatory drugs to treat allergic conditions in man. PMID- 15783134 TI - Leucocyte migration and nitroblue tetrazolium assay in Nigerian children with bacteremia and malaria parasitemia. AB - The prevalence of malaria parasitemia, bacteremia, certain hematological parameters, leucocyte migration index and nitroblue tetrazolium dye reduction were determined in 147 Nigerian children (4.24+/-2.88 years of age). Sixty (40.8%), 28(19.1%) and 26(17.7%) had malaria parasitemia only, bacteremia only and both malaria parasitemia and bacteremia, respectively. Four genera of bacteria, i.e E. coli, Proteus, Staphylococcus and Salmonella, were detected in subjects with both malaria parasitemia and bacteremia. The 4 bacterial genera and Klebsiella were detected in subjects with bacterial infection only. P. falciparum (68%), P. malariae (25%) and P. ovale (7%) were the species of malaria parasites identified in our subjects. Bacteremia was most prevalent in subjects with hemoglobin AA (HbAA) (60.7%) followed by HbAC (21.45%). Packed cell volume (PCV) and Hb concentration were similar in all groups but mean counts of red blood cells (RBC) and white blood cells (WBC) were statistically significantly lower in subjects with malaria parasites only compared to the controls. Leucocyte migration was significantly reduced in children with bacteremia only or both malaria parasitemia and bacteremia compared to controls, while the nitroblue tetrazolium assay was significantly reduced in children with bacteremia only. It may be concluded that malaria parasitemia significantly affects both leucocyte migration and nitroblue tetrazolium assay. PMID- 15783135 TI - Molecular cloning and characterization of a glutathione S-transferase encoding gene from Opisthorchis viverrini. AB - An adult stage Opisthorchis viverrini cDNA library was constructed and screened for abundant transcripts. One of the isolated cDNAs was found by sequence comparison to encode a glutathione S-transferase (GST) and was further analyzed for RNA expression, encoded protein function, tissue distribution and cross reactivity of the encoded protein with other trematode protein counterparts. The cDNA has a size of 893 bp and encodes a GST of 213 amino acids length (OV28GST). The most closely-related GST of OV28GST among those published for trematodes is a 28 kDa GST of Clonorchis sinensis as shown by multiple sequence alignment and phylogenetic analysis. Northern analysis of total RNA with a gene-specific probe revealed a 900 nucleotide OV28GST transcriptional product in the adult parasite. Through RNA in situ hybridization OV28GST RNA was detected in the parenchymal cells of adult parasites. This result was confirmed by immunolocalization of OV28GST with an antiserum generated in a mouse against bacterially-produced recombinant OV28GST. Both, purified recombinant and purified native OV28GST were resolved as 28 kDa proteins by SDS-PAGE. Using the anti-recOV28GST antiserum, no or only weak cross-reactivity was observed in an immunoblot of crude worm extracts against the GSTs of Schistosoma mansoni, S. japonicum, S. mekongi, Eurytrema spp. and Fasciola gigantica. The enzyme activity of the purified recombinant OV28GST was verified by a standard 1-chloro-2, 4-dinitrobenzene (CDNB) based activity assay. The present results of our molecular analysis of OV28GST should be helpful in the ongoing development of diagnostic applications for opisthorchiasis viverrini. PMID- 15783136 TI - Distinct immunologic properties of Penicillium marneffei yeasts obtained from different in vitro growth conditions. AB - A dimorphic fungus Penicillium marneffei is a causative agent of penicilliosis, a life-threatening disseminated disease in immunocompromised hosts predominantly found in southeast Asia and southern China. P. marneffei is the only known Penicillium that possesses a dimorphic characteristic. Since it is difficult to produce large amount of P. marneffei yeasts in vivo for experimentation purpose, yeast cells were produced in different in vitro conditions as alternatives. We interested in investigating the immunologic properties of yeast cells from different culture preparations. It was found that yeast cells obtained from brain heart infusion broth and Sabouraud dextrose broth did not resemble those resided in clinical specimens. A solution of 1% peptone, on the other hand, could induce a direct conidial transition into fission yeasts. Ability of yeast cells in each preparation to activate macrophages was determined by analyzing surface expression of CD40 and CD86 co-stimulatory molecules after two days of co cultivation. Every P. marneffei yeast cell preparation demonstrated such ability. However, the ones from Sabouraud dextrose broth seemed to induce less phagocytosis. Additionally, although distinct antigenic profiles and lack of conformity in antigenic expression were observed among yeast cells from different culture conditions, most major immunogenic bands were present when Western analysis was performed using polyclonal antisera from penicilliosis patients. The results of the study raise attention on immunological and biochemical characteristics of P. marneffei yeasts if such preparations are to be used in future laboratory investigations. PMID- 15783137 TI - Changing hepatitis A epidemiology and the need for vaccination in Korea. AB - Hepatitis A is a vaccine-preventable disease with 1.5 million people infected world-wide annually. Improvement in the socio-economic status and general public health measures of Asian countries over the last 20 years has led to a shift in the seroprevalence of hepatitis A in many of these countries. In Korea, like in many other developed countries, this lowered endemicity has caused an upward shift in the average age of infection, resulting in larger numbers of individuals at risk of clinically significant hepatitis A infection. Sporadic outbreaks increase the public health burden of the disease. Inactivated hepatitis A vaccines are an effective prevention measure and have been shown to be safe, efficacious and well-tolerated in Korean children. Given this changing epidemiology of the disease and the associated increase in morbidity, vaccination of young children who are not immune, as well as other high risk groups, should be recommended. PMID- 15783138 TI - [Differences in disciplines, complementarity and nursing]. PMID- 15783139 TI - [Ulcer, scarring, what do we know?]. PMID- 15783140 TI - [Wounds in palliative nursing]. PMID- 15783141 TI - [Caring fo infection in chronic ulcers]. PMID- 15783142 TI - [How should one change the practice of nurses in wounds and scars?]. PMID- 15783143 TI - [Diet education is insufficient in cardiac care]. PMID- 15783144 TI - [Insufficiency in cardiac care, a new domain in therapeutic education]. PMID- 15783145 TI - [The role of the liberal nurses in the care of the patient with cardiac insufficiency]. PMID- 15783146 TI - [Diet and cardiology]. PMID- 15783147 TI - [Evaluation of arterial pressure]. PMID- 15783148 TI - [Marseille and the plague, a story of rebounding]. PMID- 15783149 TI - Going head-to-head with the PM. PMID- 15783150 TI - A voice from older people's nursing. PMID- 15783151 TI - Moments that make nursing worthwhile. PMID- 15783152 TI - A district nursing renaissance. PMID- 15783153 TI - Children's Trust comes of age. PMID- 15783154 TI - Providing older patients with holistic health promotion. AB - Health promotion strategies for older people should focus on positive activities, taking a holistic approach and accounting for the needs of the individual. This article outlines the development of a nurse-led project that aims to meet the health promotion need of this patient group. PMID- 15783155 TI - Tinnitus. PMID- 15783156 TI - Nicotine replacement therapy. PMID- 15783157 TI - A cognitive sketch pad to record thoughts of psychiatric patients. AB - The purpose of a cognitive sketch pad is to facilitate thoughts that allow patients to be engaged with their nurse and with activities of daily living. This recording of thoughts is a way of finding areas of agreement that will move patients' care towards recovery. PMID- 15783158 TI - The role of nurses in managing patients with heart failure. AB - Heart failure is a clinical syndrome caused by a reduction in the heart's ability to pump blood around the body. The prevalence of heart failure due to coronary heart disease (CHD) is increasing, despite the declining overall mortality from CHD. This is thought to be due to both an ageing population and to more people surviving acute heart attacks but then experiencing residual left ventricular dysfunction (Department of Health, 2002). The National Service Framework for Coronary Heart Disease (DoH, 2002) sets out clear standards for the treatment of patients with heart failure. PMID- 15783160 TI - Helping to understand the pathophysiology of anaemia. PMID- 15783159 TI - The role of viruses and sexual transmission in anal cancer. AB - Research has focused on the link between sexual activity, viral infection and cervical cancer. However, a parallel situation can be seen with anal cancer. Although less common than cervical cancer, anal cancer is a significant problem among certain groups. In the male homosexual population it occurs in 35 out of every 100,000 men, a figure comparable with the rate of cervical cancer in women before cervical screening programmes were instigated (Klenke and Palefsky, 2003). This article discusses the pathology, incidence and management of the disease and considers the role of viruses in its development, specifically human papilloma virus (HPV) and human immunodeficiency virus (HIV). PMID- 15783161 TI - Exploring the outer limits of nursing. PMID- 15783162 TI - Matron or Manimal? PMID- 15783163 TI - Solution structure of a beta-peptide ligand for hDM2. AB - We recently reported a beta-peptide foldamer, beta53-1, that folds into a 14 helix in aqueous solution, binds the oncoprotein hDM2 with submicromolar affinity, and potently inhibits the interaction of hDM2 with a peptide derived from the activation domain of p53 (p53AD). Here, we present the solution structure of beta53-1 in methanol. Details of the structure illustrate fundamental and novel elements of beta-peptide folding and recognition. These elements include the detailed arrangement of a complex, 14-helix-stabilizing salt bridge on one helical face, and a unique "wedge into cleft" packing interaction along a second. The structure also reveals how a subtle distortion in the beta53 1 14-helix geometry alters the presentation of its recognition epitope, rendering it particularly well suited for alpha-helix mimicry. The solution structure of beta53-1 demonstrates that well folded beta-peptide oligomers can effectively present an extended, highly variable surface that could be used as a general platform for targeting critical protein-protein interfaces. PMID- 15783164 TI - The role of chelating diamine ligands in the goldberg reaction: a kinetic study on the copper-catalyzed amidation of aryl iodides. AB - The mechanistic details of the Cu-catalyzed amidation of aryl iodides are presented. The kinetic data suggest that the diamine ligand prevents multiple ligation of the amide. The formation of an amidocuprate species external to the catalytic cycle helped to rationalize the dependence on diamine concentration and the inverse dependence on amide concentration at low diamine concentrations. The intermediacy of a Cu(I) amidate was established through both its chemical and kinetic competency. PMID- 15783165 TI - Asymmetric end-functionalization of multi-walled carbon nanotubes. AB - Asymmetric end-functionalization of carbon nanotubes was achieved by sequentially floating a substrate-free aligned carbon nanotube film on two different photoreactive solutions with only one side of the nanotube film being contacted with the photoreactive solution and exposed to UV light each time. The resultant nanotubes with different chemical reagents attached onto their opposite tube-ends should be very useful for site-selective self-assembling of carbon nanotubes into many novel functional structures for various potential applications. PMID- 15783166 TI - Switching of intramolecular charge transfer in cruciforms: metal ion sensing. AB - Three different cruciforms containing the 1,4-bis-4'-pyridylethynyl-2,5 distyrylbenzene or the 1,4-bis-phenylethynyl-2,5-distyrylbenzene unit were synthesized and investigated with respect to their metal sensing properties. Upon addition of metal cations to these cruciforms, either a bathochromic or hypsochromic shift in emission and absorption is observed. The shift depends on whether the metal coordinates preferentially to the pyridine or to the dibutylaniline branches of the cruciforms. The three cruciforms were exposed to a selection of metal cations in dichloromethane, and their emission was examined. The cruciforms show differential sensing of metal cations, that is, different metals can be specifically discerned upon exposure to three cruciforms but not when exposed to only one. The system can discern calcium from magnesium cations and silver from mercury or lithium cations. PMID- 15783167 TI - Ligand effect on the growth and the digestion of Co nanocrystals. AB - The reaction product of cobalt carbonyl decomposition depends on the concentration of the oleic acid ligand. With a low concentration of ligand, nanocrystals nucleate and grow to large ferromagnetic particles through the process of Ostwald ripening and coalescence coarsening. With a high concentration of ligand, stable cluster complexes are formed. Addition or removal of ligand from the reaction products can interchange the formation of cluster complexes and nanocrystals. PMID- 15783168 TI - Amphiphilic polymethacrylate derivatives as antimicrobial agents. AB - We have investigated the structure-activity relationship of cationic amphiphilic polymethacrylate derivatives in antimicrobial and hemolytic assays. The polymers were prepared by radical copolymerizations of N-(tert-butoxycarbonyl)aminoethyl methacrylate and butyl methacrylate in the presence of methyl 3 mercaptopropionate as a chain transfer agent to give precursor polymers protected with a tert-butoxycarbonyl (Boc) group. Subsequent treatment of the Boc-protected polymers with TFA affords the desired cationic random copolymers. We examined antimicrobial and hemolytic activities of a series of polymers having a wide range of mole percentage of butyl groups (0-60%) in three different molecular weight (MW) ranges. The smallest polymers (MW < 2000) showed the lowest MIC and reduced hemolytic activity compared to that of the higher MW ones. In addition, polymers containing a high percentage of butyl groups are less selective for bacterial cells than their less hydrophobic counterparts. PMID- 15783169 TI - Facile modification of collagen directed by collagen mimetic peptides. AB - Recent widespread interest in the development of engineered tissue and organ replacement therapies has prompted demand for new approaches to immobilize exogenous components to natural collagen. Chemical coupling of synthetic moieties to amino acid side chains has been commonly practiced for such purposes; however, such coupling reactions are difficult to control on large proteins and are generally not conducive to modifying integrated collagen scaffolds that contain live cells and tissues. As an alternative to the conventional "covalent" modification method, we have developed a novel "physical" modification technique that is based on collagen's native ability to associate into a triple-helical molecular architecture. Here, we present a finding that collagen mimetic peptides (CMPs) of sequence -(Pro-Hyp-Gly)x- exhibit strong affinity to both native and gelatinized type I collagen under controlled thermal conditions. We also show that the cell adhesion characteristics of collagen can be readily altered by applying a poly(ethylene glycol)-CMP conjugate to a prefabricated collagen film. PMID- 15783170 TI - Unprecedented control over polymerization of n-hexyl isocyanate using an anionic initiator having synchronized function of chain-end protection. AB - The living anionic polymerization of isocyanates carried out using conventional initiators is associated with side reactions due to rapid initiation rates as well as back-biting by the growing chain, resulting in a lack of control on the molecular weight (MW) and molecular weight distribution (MWD) of the polymers. Successful control over the reaction was possible by using additives that could prevent back-biting. We find an initiator in sodium benzanilide (Na-BA), which has a slow initiation rate combined with additive function, so that use of an external additive is eliminated. The initiator has resulted in polymers with high yields and an unprecedented control over the MW and MWD. It is possible to introduce a number of functionalities at the termini of the polymer by using Na BA derivatives as well as suitable terminating agents, leading to macromonomer, reactive and chiral polymers, and chiral macromonomer in approximately 100% yields. In the process, the finding has expanded the scope of polyisocyanates in diverse applications. PMID- 15783171 TI - Insight into binding of alkanes to transition metals from NMR spectroscopy of isomeric pentane and isotopically labeled alkane complexes. AB - Alkane complexes of the type Cp'Re(CO)2(alkane) (Cp' = cyclopentadienyl or (isopropyl)cyclopentadienyl; alkane = isotopomers of n-pentane and cyclopentane) have been characterized using NMR spectroscopy following photolysis of Cp'Re(CO)3 in the appropriate alkane at 163-193 K. In the case of n-pentane, three different complexes are observed corresponding to binding of the three different types of carbon in this alkane. ROESY NMR experiments indicate that these isomeric complexes are slowly interconverting intramolecularly at 173 K. The order of the energetically preferred site of coordination is methylene (C2) approximately central methylene (C3) > methyl (C1) but with a spread of <0.2 kcal mol-1. Isotopic perturbation of resonance (IPR) experiments, conducted on several isotopomers of (i-PrCp)Re(CO)2(1-pentane), showed a large shielding of the 1H NMR chemical shift of the proton in a bound CHD2 moiety (delta -3.62) and CH2D (delta -2.64) compared with that of a bound CH3 moiety (delta -1.99). Likewise, the value of 1JCH for the coordinated methyl group of isotopomers of (i PrCp)Re(CO)2(1-pentane) reduces in the order CH3 > CH2D > CHD2. This suggests that the alkane coordinates in an eta2-C,H fashion with a rapid exchange of bound hydrogen or deuterium within a methyl or methylene group, and that binding of a hydrogen atom is preferred over a deuterium by an amount of 0.23 +/- 0.03 kcal mol-1. PMID- 15783172 TI - Quantifying adhesion bond parameters to distinguish interactions of hydrophilic and hydrophobic blocks of polystyrene-poly-2-vinylpyridine with a silicon nitride surface. AB - An analysis of the loading rate dependence of the forces required to rupture an AFM tip from a block copolymer surface has provided insight into the structure of the surface-macromolecule contact, differentiation of the block contacting the tip, a measure of the polymer-surface binding energy, and the rigidity of the contact. Polystyrene-poly-2-vinylpyridine block copolymers were studied adsorbing to silicon nitride. Polystyrene makes stiff van der Waals contact with the silicon nitride surface in aqueous solution, while the bond of poly-2 vinylpyridine to the surface is more flexible and may involve a bridging water. PMID- 15783173 TI - Enantioselective alkenylation and phenylation catalyzed by a chiral CuF complex. AB - A new method for CuF-catalyzed alkenylation and phenylation of aldehydes and an activated ketone using air- and moisture-stable alkenylsilanes and phenylsilane as a nucleophile is described. This methodology was extended to highly enantioselective catalytic alkenylation and phenylation using DTBM-SEGPHOS as a chiral ligand. Substrate generality is broad, and an alkenylsilane with a long alkyl chain and an internal alkenylsilane can be also used as a nucleophile. The key to success partly involves the accelerated regeneration of reactive alkenylcopper and phenylcopper through transmetalation from the silylated nucleophiles, and stabilization of the reactive copper reagents, both of which are effected by the diphosphine ligands. PMID- 15783174 TI - (+)-Myristinin A, a naturally occurring DNA polymerase beta inhibitor and potent DNA-damaging agent. AB - The first stereoselective total synthesis of the naturally occurring flavan myristinin A has been accomplished, as well as its biochemical evaluation. This synthesis verified the structural assignment and allowed for the determination of the absolute stereochemistry. Myristinin A exhibits biochemical activity both as a potent DNA-damaging agent and DNA polymerase beta inhibitor. Relaxation of supercoiled plasmid DNA was observed at picomolar concentrations, in addition to inhibition of polymerase beta at low micromolar concentrations. PMID- 15783175 TI - A stable N-heterocyclic carbene with a diboron backbone. AB - The synthesis of a novel five-membered inorganic ring, a stable N-heterocyclic carbene with a diboron backbone, is reported. A pentacarbonyltungsten complex containing the new carbene is also described. Spectroscopic evidence indicates that the sterically encumbered carbene is a better sigma-donor than classical N heterocyclic analogues, demonstrating the significant influence of the inorganic backbone on the coordinating properties of the carbon ligand. Crystal structures have been determined for an iminium precursor, the free carbene, and the tungsten complex. PMID- 15783176 TI - (SR/RS)-cyclohexanyl PNAs: conformationally preorganized PNA analogues with unprecedented preference for duplex formation with RNA. AB - PNA oligomers H-GTAGATCAT-lys-NH2 with cis-(1S,2R/1R,2S)-cyclohexyl-T (III) in the backbone form PNA:RNA duplexes with Tm approximately 30-50 degrees C higher than that of PNA:DNA duplexes. In comparison, cis-(1S,2R/1R,2S)-cyclopentyl PNA-T (IV) form highly stable duplexes with both RNA and DNA without discrimination. PMID- 15783177 TI - 19F NMR characterization of the thermodynamics and dynamics of the acid-alkaline transition in a reconstituted sperm whale metmyoglobin. AB - A 19F NMR study on the acid-alkaline transition in sperm whale myoglobin reconstituted with a perfluoromethyl heme, 13,17-bis(2-carboxylatoethyl)-3,8 diethyl-2,12,18-trimethyl-7-trifluoromethylporphyrinatoiron(III), demonstrated that the thermodynamics of the transition is predominantly controlled by the stability of acidic form. PMID- 15783179 TI - RuO2-loaded beta-Ge3N4 as a non-oxide photocatalyst for overall water splitting. AB - Germanium nitride beta-Ge3N4 dispersed with RuO2 nanoparticles is presented as the first example of a non-oxide photocatalyst for the stoichiometric decomposition of H2O into H2 and O2. All of the successful photocatalysts developed for overall water splitting over the past 30 years have been based on oxides of metals. The discovery of a non-oxide photocatalyst, such as nitrides and oxynitrides, achieving the same function is therefore expected to stimulate research on non-oxide photocatalysts. New opportunities for progress in the development of visible light-driven photocatalysis can thus be expected, as the higher valence band positions of metal nitrides compared to the corresponding metal oxides provide narrower band gaps, which are suitable for visible light activity. PMID- 15783178 TI - Fourier transform infrared characterization of the azido complex of methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath). AB - The azido complex formed in oxidized methane monooxygenase from Methylococcus capsulatus (Bath) was investigated with resonance Raman and FTIR techniques. These experiments show the presence of a nuas(NNN) at approximately 2077 cm-1 which splits to two components at 2059 and 2073 cm-1 with 15N14N2. The vibrational data are assigned to an azido complex bound terminally to one iron(III) at the diiron center. When the azido complex is illuminated at 15 K, a new nuas(NNN) is observed at 2136 cm-1 which is assigned to a photodissociated HN3 within the substrate pocket. We propose a model where an aqua ligand engages a hydrogen bond interaction with the 1N atom of the azido group and acts as at a proton donor during the photolysis process. PMID- 15783180 TI - A ruthenium-dihydrogen putative intermediate in ketone hydrogenation. AB - The compound fac-[Ru((R)-BINAP)(H)(2-PrOH)3]+ (6) (BINAP = 2,2' bis(diphenylphosphino)-1,1'-binaphthyl) reacts with (R,R)-dpen (dpen = 1,2 diphenylethylenediamine) under H2 at -60 degrees C in 2-PrOH-d8/CD2Cl2 to generate the cationic dihydrogen putative intermediate trans-[Ru((R) BINAP)(H)(eta2-H2)((R,R)-dpen)]+ (2') without H-D exchange between the hydrogen ligands and the solvent. A 1H NMR study concludes that the dihydrogen ligand in 2' does not protonate 2-PrOH to a catalytically significant extent, and that 2' requires an added base or hydride source to be an active catalyst. PMID- 15783181 TI - Highly enantioselective Friedel-Crafts alkylations of pyrroles and indoles with alpha'-hydroxy enones under Cu(II)-simple bis(oxazoline) catalysis. AB - Remarkably high and regular enantioselectivities are obtained in Friedel-Crafts alkylation reactions involving alpha'-hydroxy enone templates and Cu(II) bis(oxazoline) complexes as catalysts. The simple elaboration of adducts provides a route to enantioenriched aldehydes, carboxylic acids, and ketones containing the pyrrole and indole frameworks. PMID- 15783182 TI - Catalytic coupling of haloolefins with anilides. AB - A strategy in which C-H activation reactions promoted by Pd(II) have been combined with beta-heteroatom elimination to create a catalytic cycle achieving the arylation of haloacrylates is reported. The catalytic cycle can be subdivided into four parts: (1) C-H activation; (2) the functionalization step, migratory insertion of the olefin into a metal-carbon bond; (3) beta-heteroatom elimination; and (4) exchange of metal halide (if X = halogen) for a less coordinating anion. In this catalytic cycle, the oxidation state of the metal does not change, and an oxidant is not required. The method is more functional group tolerant compared with the existing alkene-arene coupling methods based on electrophilic C-H activation. PMID- 15783183 TI - Cell-permeable near-infrared fluorogenic substrates for imaging beta-lactamase activity. AB - This communication describes a design of cell-permeable near-infrared fluorogenic substrates for imaging beta-lactamase expression in living mammalian cells. This design is based on fluorescence energy transfer resonance and utilizes a peracetylated d-glucosamine to facilitate the transport of the near-infrared probe across cell membranes. This new type of fluorogenic probe may also be applied to image gene expression in living animals. PMID- 15783184 TI - Liquid-core capsules via interfacial polymerization: a free-radical analogy of the nylon rope trick. AB - Liquid-core capsules have wide-ranging applications in the high-efficiency encapsulation and controlled release of drugs, dyes, enzymes, and other substrates. Their great utility has driven the rapid development of various preparation techniques. However, there remains no convenient technique for the preparation of submicrometer liquid-core capsules with shell thicknesses less than 100 nm. Here, we demonstrate a new interfacial free-radical polymerization approach for the straightforward preparation of liquid-core polymer capsules. Conceptually, this interfacial free-radical polymerization is analogous to the classical "nylon rope trick" wherein hydrophobic and hydrophilic monomers alternately copolymerize to constrain the polymerization at interfaces, but its free-radical mechanism allows precise control of initiation, which makes it possible to finely disperse the immiscible phases prior to polymerization. PMID- 15783185 TI - Electronic structures and properties of twisted polyacenes. AB - The effects of twisting on the electronic structures and properties of polyacenes were studied computationally using DFT methods. Singlet-triplet and HOMO-LUMO gaps and vertical S0-S1 transition energies are marginally affected as a function of end-to-end twist angle. The large twist induced by the phenyl substituents in 9,10,11,20,21,22-hexaphenyltetrabenzo[a,c,l,n]pentacene has little influence on its electronic structure. PMID- 15783186 TI - A stable single piece of unimolecularly pi-stacked porphyrin aggregate in a thixotropic low molecular weight gel: a one-dimensional molecular template for polydiacetylene wiring up to several tens of micrometers in length. AB - An amide-type copper porphyrin gelator having alkyldiacetylene tethers gives very transparent gel in Decalin, and the gel shows unique thixotropic behavior. One dimensional aggregates generated in the gel act as templates for unimolecularly segregated polydiacetylene wiring upon UV irradiation on the surface. The polymerization along the aggregation goes to at least several micrometers long. PMID- 15783187 TI - Monitoring the growth of polyoxomolybdate nanoparticles in suspension by flow field-flow fractionation. AB - We follow the evolution of polyoxomolybdate nanoparticles in suspensions derived from the keplerate (NH4)42[MoVI72MoV60O372(CH3CO2)30(H2O)72].ca..300H2O.ca..10CH3CO2NH4 ({Mo132}) by flow field-flow fractionation (FlFFF) to monitor the particle-size distribution in situ, atomic force and high-resolution transmission electron microscopy (AFM, SEM, and HRTEM) to confirm particle sizes, inductively coupled plasma-optical emission spectrometry (ICP-OES) to determine the Mo content of the FlFFF separated fractions, and UV/visible spectroscopy to confirm the identity of the species in suspension. We observe the formation of 3-75-nm polyoxomolybdate particles in suspension and the dynamic growth of {Mo132} crystals. PMID- 15783188 TI - Bulky diarylammonium arenesulfonates as selective esterification catalysts. AB - More environmentally benign alternatives to current chemical processes, especially large-scale, fundamental reactions such as ester condensations, are highly desirable for many reactions. Bulky diarylammonium pentafluorobenzenesulfonates and tosylates serve as extremely active dehydration catalysts for the ester condensation reaction of carboxylic acids with equimolar amounts of sterically demanding alcohols and acid-sensitive alcohols. Typically, the esterification reaction is performed in heptane by heating at 80 degrees C in the presence of 1 mol % of the catalyst without removing water. Esterification with primary alcohols proceeds without solvents even at room temperature. Furthermore, 4-(N-mesitylamino)polystyrene resin-bound pentafluorobenzenesulfonate can be recycled more than 10 times without activity loss. PMID- 15783189 TI - Heptamethine cyanine dyes with a large stokes shift and strong fluorescence: a paradigm for excited-state intramolecular charge transfer. AB - New heptamethine cyanine dyes with an alkylamino group at the central position were found to exhibit a large Stokes shift (>140 nm) and strong fluorescence. They were suggested to be a new paradigm for excited-state intramolecular charge transfer (ICT). The configuration change of the bridgehead amine accompanying ICT was investigated in different viscosity and pH media. PMID- 15783190 TI - Synthesis and circular dichroism of tetraarylporphyrin-oligonucleotide conjugates. AB - The preparation and circular dichroic (CD) studies of self-complimentary 8-mer DNA sequences with a porphyrin at the 3' end are presented. Electronic interaction between the two porphyrins (the interchromophoric distance is in the range of 28-40 A), attached to both ends of the double-stranded helix, gives rise to a long-range exciton-coupled CD in the visible region (400-450 nm). The porphyrin chromophores act as sensitive probes of geometrical changes in the DNA backbone and sensitively reflect the double-strand to single-strand transition. This study demonstrates the possibility of using exciton-coupled porphyrin CDs for conformational studies of DNA. PMID- 15783191 TI - A simple glycol nucleic acid. AB - A glycol nucleic acid (GNA) with an acyclic propylene glycol phosphodiester backbone forms stable antiparallel duplexes following the Watson-Crick base pairing rules. PMID- 15783192 TI - Design of an intein that can be inhibited with a small molecule ligand. AB - Protein splicing is a process in which an intervening sequence, the intein, catalyzes its own excision out of a larger polypeptide precursor by joining the flanking sequences, the exteins, with a native peptide bond. Inteins are almost completely promiscuous toward the nature of their extein sequences and can be inserted into virtually any host protein. The intein-mediated formation of a peptide bond between two polypeptides offers great potential to modulate protein structure and, hence, protein function on the post-translational level. In this work, we report the design of an intein that can be inhibited by the addition of a specific small molecule ligand. Our design strategy involved the generation of a trans-splicing intein, in which the intein domain is split into two-halves that are located on two separate polypeptides, each joined with the respective N- or C terminal extein. To turn these fragments into an active intein with an incorporated "off" switch, each was fused at its newly created terminus with the F36M mutant of FKBP12, referred to as the FM domain. The F36M substitution was reported to effect a homodimerization of the usually monomeric FKBP12 protein; however, addition of the small molecule ligand, rapamycin, or synthetic derivatives thereof leads to a dissociation of the dimer. This phenomenon was exploited by first reconstituting the active intein on the basis of FM domain dimerization. Second, addition of the small molecule ligand prevented formation of the active intein complex and inhibited protein trans-splicing. This intein exhibited unexpected kinetic properties and provides a new and potentially very general means to control protein function on the post-translational level. PMID- 15783194 TI - Sulfoxide carbon-sulfur bond activation. AB - The alkynylsulfoxide, TMSCCSO(p-tolyl) (TMS = trimethylsilyl, tolyl = C6H4Me), undergoes reaction with (eta5-C5H5)Co(PPh3)2 at room temperature to give the cobaltosulfoxide complex, (C5H5)Co(PPh3)(eta1-CCTMS)[eta1-(S)-SO(p-tolyl)], which was characterized by X-ray crystallography. Exposure of this cobaltosulfoxide complex to oxygen gas leads to the formation of the corresponding metallosulfone complex, (C5H5)Co(PPh3)(eta1-CCTMS)[eta1-(S)-SO2(p-tolyl)], which was characterized by X-ray crystallography. Alternatively, in solution at room temperature, the metallosulfoxide is converted to a 1:4 mixture of the equatorial equatorial and equatorial-axial bridging cobalt-thiolato dimers, {(C5H5)Co[mu-S(p tolyl)]}2, respectively. The equatorial-equatorial isomer was characterized by X ray crystallography. PMID- 15783193 TI - Dioxygen activation and catalytic aerobic oxidation by a mononuclear nonheme iron(II) complex. AB - We have used dioxygen, not artificial oxidants such as peracids, iodosylarenes, and hydroperoxides, in the generation of a mononuclear nonheme oxoiron(IV) complex, [Fe(IV)(TMC)(O)]2+ (TMC = 1,4,8,11-tetramethyl-1,4,8,11 tetraazacyclotetradecane), from its corresponding Fe(II) complex, [Fe(TMC)(CF3SO3)2]. The formation of oxoiron(IV) species by activating dioxygen was markedly dependent on iron(II) complexes and solvents, and this observation was interpreted with the electronic effect of iron(II) complexes on dioxygen activation to form oxoiron(IV) species. A catalytic aerobic oxidation of organic substrates was demonstrated in the presence of the [Fe(TMC)]2+ complex. By carrying out 18O-labeled water experiment, we were able to conclude that the oxidation of organic substrates was mediated by an oxoiron(IV) intermediate, not by a radical type of autoxidation process. PMID- 15783195 TI - From CO oxidation to CO2 activation: an unexpected catalytic activity of polymer supported nanogold. AB - A simple, clean, safe, and reproducible catalyst system, polymer-supported nanogold, was successfully developed for the fixation of CO2 to cyclic carbonate and for the carbonylation of amines to disubstituted ureas with unprecedented catalytic activity (TOF > 50 000 mol/mol/h and TOFP approximately 3000 mol/mol/h, respectively). To the best of our knowledge, it was the first to report that nanogold catalysts have exclusive catalytic activity for activation of carbon dioxide, and that the catalytic activity of the polymer-immobilized nanogold catalysts could be controlled by the particle size of the nanogold. PMID- 15783196 TI - Rapid fixation of methylene chloride by a macrocyclic amine. AB - A simple macrocyclic amine is alkylated by methylene chloride to give a quaternary ammonium chloride salt. When methylene chloride is the solvent, the reaction exhibits pseudo-first-order kinetics, and the reaction half-life at 25.0 degrees C is 2.0 min. The reaction half-life for a structurally related, acyclic amine is approximately 50 000 times longer. Detailed calculations favor a mechanism where the methylene chloride associates with the macrocycle to form an activated prereaction complex. The macrocyclic nitrogen subsequently attacks the methylene chloride with a classic SN2 trajectory, and although the carbon chlorine bond breaks, the chloride leaving group does not separate from the newly formed cationic macrocycle, such that the product is a tightly associated ion pair. X-ray crystal structures of the starting amine and the product salt, as well as kinetic data, support this mechanism. PMID- 15783197 TI - Ruthenium-catalyzed regioselective 1,3-methylene transfer by cleavage of two adjacent sigma-carbon-carbon bonds: an easy and selective synthesis of highly substituted benzenes. AB - We report a new ruthenium-catalyzed 6-endo-dig cyclization of 6,6 cycloalkylidenyl-3,5-dien-1-ynes, which produces highly substituted benzenes with considerable structural reorganization. In this process, we observe a regioselective 1,3-methylene migration via extrusion from a cycloalkylidenyl ring, in addition to a regiocontrolled 1,2-alkyl migration. This cyclization provides an easy and convenient synthesis of complex benzenes bearing various different substituents. PMID- 15783198 TI - Graphitized pitch-based carbons with ordered nanopores synthesized by using colloidal crystals as templates. AB - A highly graphitized ordered nanoporous carbon (ONC) was synthesized by using commercial mesophase pitch as carbon precursor and siliceous colloidal crystal as template. Since silica colloids of different sizes (above 6 nm) and narrow particle size distribution are commercially available, the pore size tailoring in the resulting ONCs is possible. PMID- 15783199 TI - Dual emission from a family of conjugated dinuclear Ru(II) complexes. AB - Dual emission is observed from a family of simple acetylene-linked dinuclear RuII complexes, where two MLCT excited states coexist at room temperature and in fluid solution. This unique behavior is attributed to a specific substitution pattern on the bridging ligand and provides the opportunity to investigate the structural and electronic features that result in decoupling of standard nonradiative decay pathways. PMID- 15783200 TI - @-tides as reporters for molecular associations. AB - The 1,6-dihydro-3(2H)-pyridinone unit is an amino acid surrogate that favors the extended beta-strand conformation when incorporated in an oligopeptide ("@-tide") strand. We now report that the circular dichroism (CD) signature of the vinylogous amide in the @-unit is sensitive to conformation in organic and aqueous solvents and, therefore, is useful as a quantitative measure of @-tide association and folding processes that involve this moiety. Moreover, this method can be employed in the micromolar concentration range, which is not readily accessible using other techniques. Measurements of @-tide dimerization and beta hairpin folding equilibria not only demonstrate the utility and generality of this approach but also provide a way to quantify amino acid side chain-side chain interactions relevant to beta-sheet stability. PMID- 15783201 TI - Quadruplex formation by a guanine-rich PNA oligomer. AB - A guanine-rich PNA dodecamer having the sequence H-G4T4G4-Lys-NH2 (G-PNA) hybridizes with a DNA dodecamer of homologous sequence to form a four-stranded quadruplex (Datta, B.; Schmitt, C.; Armitage, B. A. J. Am. Chem. Soc. 2003, 125, 4111-4118). This report describes quadruplex formation by the PNA alone. UV melting curves and fluorescence resonance energy transfer experiments reveal formation of a multistranded structure stabilized by guanine tetrads. The ion dependency of these structures is analogous to that reported for DNA quadruplexes. Electrospray ionization mass spectrometry indicates that both dimeric and tetrameric quadruplexes are formed by G4-PNA, with the dimeric form being preferred. These results have implications for the use of G-rich PNA for homologous hybridization to G-rich targets in chromosomal DNA and suggest additional applications in assembling quadruplex structures within lipid bilayer environments. PMID- 15783202 TI - Reaction intermediates of quinol oxidation in a photoactivatable system that mimics electron transfer in the cytochrome bc1 complex. AB - Current competing models for the two-electron oxidation of quinol (QH2) at the cytochrome bc1 complex and related complexes impose distinct requirements for the reaction intermediate. At present, the intermediate species of the enzymatic oxidation process have not been observed or characterized, probably due to their transient nature. Here, we use a biomimetic oxidant, excited-state Ru(bpy)2(pbim)+ (bpy=2,2'-dipyridyl, pbim=2-(2-pyridyl)benzimidazolate) in an aprotic medium to probe the oxidation of the ubiquinol analogue, 2,3-dimethoxy-5 methyl-1,4-benzoquinol (UQH2-0), and the plastoquinol analogue, trimethyl-1,4 benzoquinol (TMQH2-0), using time-resolved and steady-state spectroscopic techniques. Despite its simplicity, this system qualitatively reproduces key features observed during ubiquinol oxidation by the mitochondrial cytochrome bc1 complex. Comparison of isotope-dependent activation properties in the native and synthetic systems as well as analysis of the time-resolved direct-detection electron paramagnetic resonance signals in the synthetic system allows us to conclude that (1) the initial and rate-limiting step in quinol oxidation, both in the biological and biomimetic systems, involves electron and proton transfer, probably via a proton-coupled electron-transfer mechanism, (2) a neutral semiquinone intermediate is formed in the biomimetic system, and (3) oxidation of the QH*/QH2 couple for UQH2-0, but not TMQH2-0, exhibits an unusual and unexpected primary deuterium kinetic isotope effect on its Arrhenius activation energy (DeltaGTS), where DeltaGTS for the protiated form is larger than that for the deuterated form. The same behavior is observed during steady-state turnover of the cyt bc1 complex using ubiquinol, but not plastoquinol, as a substrate, leading to the conclusion that similar chemical pathways are involved in both systems. The synthetic system is an unambiguous n=1 electron acceptor, and it is thus inferred that sequential oxidation of ubiquinol (by two sequential n=1 processes) is more rapid than a truly concerted (n=2) oxidation in the cyt bc1 complex. PMID- 15783203 TI - Solution structure of Vanabin2, a vanadium(IV)-binding protein from the vanadium rich ascidian Ascidia sydneiensis samea. AB - Ascidians belonging to the suborder Phlebobranchia are known to accumulate high levels of a transition metal, vanadium, in their blood cells, called vanadocytes, although the mechanism for this biological phenomenon remains unclear. Recently, we identified vanadium(IV)-binding proteins, designated as Vanabins, from vanadium-accumulating ascidians. Here, we report the first 3D structure of Vanabin2 from an ascidian, Ascidia sydneiensis samea, in an aqueous solution. The structure revealed a novel bow-shaped conformation, with four alpha-helices connected by nine disulfide bonds. There are no structural homologues reported so far. The 15N heteronuclear single-quantum coherence (HSQC) perturbation experiments of Vanabin2 indicated that vanadyl cations, which are exclusively localized on the same face of the molecule, are coordinated by amine nitrogens derived from amino acid residues such as lysines, arginines, and histidines, as suggested by the electron paramagnetic resonance (EPR) results. The present NMR studies provide information that will contribute toward elucidating the mechanism of vanadium accumulation in ascidians. PMID- 15783204 TI - RNA structure analysis at single nucleotide resolution by selective 2'-hydroxyl acylation and primer extension (SHAPE). AB - The reactivity of an RNA ribose hydroxyl is shown to be exquisitely sensitive to local nucleotide flexibility because a conformationally constrained adjacent 3' phosphodiester inhibits formation of the deprotonated, nucleophilic oxyanion form of the 2'-hydroxyl group. Reaction with an appropriate electrophile, N methylisatoic anhydride, to form a 2'-O-adduct thus can be used to monitor local structure at every nucleotide in an RNA. We develop a quantitative approach involving Selective 2'-Hydroxyl Acylation analyzed by Primer Extension (SHAPE) to map the structure of and to distinguish fine differences in structure for tRNAAsp transcripts at single nucleotide resolution. Modest extensions of the SHAPE approach will allow RNA structure to be monitored comprehensively and at single nucleotide resolution for RNAs of arbitrary sequence and structural complexity and under diverse solution environments. PMID- 15783205 TI - Hybrid QM/MM and DFT investigations of the catalytic mechanism and inhibition of the dinuclear zinc metallo-beta-lactamase CcrA from Bacteroides fragilis. AB - Based on hybrid QM/MM molecular dynamics simulation and density functional theoretical (DFT) calculations, we investigate the mechanistic and energetic features of the catalytic action of dizinc metallo-beta-lactamase CcrA from Bacteroides fragilis. The 200 ps QM/MM simulation of the CcrA enzyme in complex with nitrocefin shows that the substrate beta-lactam moiety is directed toward the active site dizinc center through the interactions of aminocarbonyl and carboxylate groups with the two active site zinc ions and the two conserved residues, Lys167 and Asn176. From the determination of the potential energy profile of a relevant enzymatic reaction model, it is found that the nucleophilic displacement reaction step proceeds with a low-barrier height, leading to the formation of an energetically favored reaction intermediate. The results also show that the high catalytic activity of the CcrA enzyme stems from a simultaneous operation of three catalytic components: activation of the bridging hydroxide nucleophile by zinc-coordinated Asp86; polarization of the substrate aminocarbonyl group by the first zinc ion; stabilization of the negative charge developed on the departing amide nitrogen by the second zinc ion. Consistent with the previous experimental finding that the proton-transfer reaction step is rate limiting, the activation energy of the second step is found to be 1.6 kcal/mol higher than that of the first step. Finally, through an examination of the structural and energetic features of binding of a thiazolidinecarboxylic acid inhibitor to the active site dizinc center, a two-step inhibition mechanism involving a protonation-induced ligand exchange reaction is proposed for the inhibitory action of a tight-binding inhibitor possessing a thiol group. PMID- 15783206 TI - A comparative study of the catalytic mechanisms of the zinc and cadmium containing carbonic anhydrase. AB - The catalytic mechanism for the conversion of carbon dioxide to hydrogen carbonate by a cadmium containing carbonic anhydrase was explored at density functional level employing two different models to simulate the active center of the enzyme. In the first model, the histidine residues around the metal ion were replaced with imidazole groups. Instead, in the second one, the simplest model was extended introducing two amino acidic residues generally present in the neighbor of enzyme and a deep water molecule. The results showed that cadmium carbonic anhydrase follows a reaction mechanism that is favored thermodynamically but not kinetically with respect to that of the most usual zinc-containing enzyme, both in a vacuum and in a protein environment. PMID- 15783207 TI - Phospholipid tubelets. AB - A novel electron microscopy specimen protocol shows that the presumed phospholipid bilayer membrane ribbons that wind helically to form the cylinders known as "tubules" are actually flattened tubes. These flattened tubes are alternatively found with a helical twist about the tube's long axis or occasionally flat with no winding or twist. Flat, cylindrically wound and axially twisted segments are routinely found along a single tube's length, and at the helically wound and axially twisted segment junctions, the chiral sense of the structure often, but not always, changes chiral sense. PMID- 15783208 TI - Guidelines in implementing enantioselective indicator-displacement assays for alpha-hydroxycarboxylates and diols. AB - Enantioselective association events between boronic acid receptors and bifunctional substrates such as alpha-hydroxycarboxylates and vicinal diols were exploited to develop enantioselective indicator-displacement assays. It was found that the sensitivity of the assay could be tuned by using either colorimetric or fluorescent indicators. Mathematical principles were established for concurrent determination of concentration and enantiomeric excess (ee) of a chiral analyte, which has been a challenge to the high-throughput analysis of chiral samples. Furthermore, experimental protocols were optimized to maximize the chiral resolution of the assay and to improve the accuracy of the ee measurements. PMID- 15783209 TI - Triplet acetylenes as synthetic equivalents of 1,2-bicarbenes: phantom n,pi state controls reactivity in triplet photocycloaddition. AB - Diaryl acetylenes, in which one of the aryl groups is either a pyridine or a pyrazine, undergo efficient triplet state photocycloaddition to 1,4 cyclohexadiene with formation of 1,5-diaryl substituted tetracyclo[3.3.0.0(2,8).0(4,6)]octanes (homoquadricyclanes). In the case of pyrazinyl acetylenes, the primary homoquadricyclane products undergo a secondary photochemical rearangement leading to diaryl substituted tricyclo[3.2.1.0(4,6)]oct-2-enes. Mechanistic and photophysical studies suggest that photocycloaddition proceeds through an electrophilic triplet excited state whereas the subsequent rearrangement to the tricyclooctenes proceeds through a singlet excited state. Chemical and quantum yields for the cycloaddition, in general, correlate with the electron acceptor character of aryl substituents but are attenuated by photophysical factors, such as the competition between the conversion of acetylene singlet excited state into the reactive triplet excited states (intersystem crossing: ISC) and/or to the radical-anion (photoelectron transfer from the diene to the excited acetylene: PET). Dramatically enhanced ISC between pi-pi S(1) state and "phantom" n,pi triplet excited state is likely to be important in directing reactivity to the triplet pathway. The role of PET can be minimized by the judicious choice of reaction conditions (solvent, concentration, etc.). From a practical perspective, such reactions are interesting because "capping" of the triple bond with the polycyclic framework orients the terminal aryl (4-pyridyl, 4-tetrafluoropyridyl, phenyl, etc.) groups in an almost perfect 60 degrees angle and renders such molecules promising supramolecular building blocks, especially in the design of metal coordination polymers. PMID- 15783210 TI - Influence of alkyl substituents on the solution- and surface-organization of hexa peri-hexabenzocoronenes. AB - Three hexa-peri-hexabenzocoronenes (HBCs) with branched, bulky alkyl substituents of different lengths in the periphery of the aromatic core have been synthesized to tune the self-association properties in solution. 1H NMR and photophysical measurements were used to probe the solution organization in comparison to the known hexa-dodecyl-substituted HBC in different solvent systems. Thermodynamic parameters for the self-association in solution, obtained by curve fitting of the concentration- and temperature-dependent NMR data using van't Hoff analysis, indicated that the self-association is an enthalpically driven process that is entropically disfavored. Photoluminescence and NMR results were both employed to determine the critical concentration where no self-association for different compounds occurred. The interactions between the molecules could be controlled by varying the nonsolvent content in the solvent mixtures, supporting the model of solvophobic effects. The spatial demand of the solubilizing side chains modulated the self-association in solution. This behavior was translated into the solution casting process, where the kinetic in addition to the thermodynamic parameters played an essential role for structure formation. The study illuminates the relationship between the solution association of HBCs and the morphology, when processed on a surface. These results are essential for the application of these materials in devices. PMID- 15783212 TI - Absolute configuration, conformation, and circular dichroism of monocyclic arene dihydrodiol metabolites: it is all due to the heteroatom substituents. AB - Absolute configurations and conformations of selected cis-1,2-dihydrodiols, isolated from bacterial enzyme-catalyzed arene dihydroxylation, have been examined by comparison of experimental and DFT-calculated CD spectra and confrontation with the results of X-ray diffraction studies in the crystalline phase. The equilibrium between the diene P and M conformers in cis-dihydrodiols is strongly dependent on the intramolecular OH-OH, OH-pi, and OH-F hydrogen bonding pattern and is crucial in determining the sign and magnitude of the long wavelength diene pi-pi transition Cotton effect. The differences originate from a dominant contribution of either P-helical (1b, X=Me) or M-helical conformers (1d, X=F), or are due to M and P low-energy conformers, both contributing a positive rotational strength (1c, X=Br). Computations show that cis-dihydrodiol 1e (X=CF3) has only one M conformer stabilized by an intramolecular O-H...F hydrogen bond. cis-Dihydrodiol 1f (X=CN) shows a Cotton effect of the sign opposite to the sense of helicity of the dominating conformer. The results of the computations highlight the inadequacy of the Diene Helicity Rule and the Allylic Chirality Rule to correlate observed Cotton effects with dihydrodiol absolute configuration. A reliable model is presented to predict the absolute configuration of substituted benzene dihydrodiol derivatives from CD spectra, based on the confrontation of DFT-computed and experimental CD spectra. For 3 alkyl derivatives, a simple noncomputational model is offered, which is based on the contributions of the allylic hydroxy groups and the diene core in P and M conformers. PMID- 15783211 TI - Total syntheses of amphidinolides T1 and T4 via catalytic, stereoselective, reductive macrocyclizations. AB - Described in this work are total syntheses of amphidinolides T1 and T4 using two nickel-catalyzed reductive coupling reactions of alkynes, with an epoxide in one case (intermolecular) and with an aldehyde in another (intramolecular). The latter was used to effect a macrocyclization, form a C-C bond, and install a stereogenic center with >10:1 selectivity in both natural product syntheses. Alternative approaches in which intermolecular alkyne-aldehyde reductive coupling reactions would serve to join key fragments were investigated and are also discussed; it was found that macrocyclization (i.e. intramolecular alkyne aldehyde coupling) was superior in several respects (diastereoselectivity, yield, and length of syntheses). Alkyne-epoxide reductive couplings were instrumental in the construction of key fragments corresponding to approximately half of the molecule of both natural products. In one case (T4 series), the alkyne-epoxide coupling exhibited very high site selectivity in a coupling of a diyne. A model for the stereoselectivity observed in the macrocyclizations is also proposed. PMID- 15783214 TI - Convergent total synthesis of gymnocin-A and evaluation of synthetic analogues. AB - The first total synthesis of gymnocin-A (1), a cytotoxic polycyclic ether isolated from a notorious red tide dinoflagellate, Karenia mikimotoi, has been accomplished. The synthesis relies heavily on the Suzuki-Miyaura cross-coupling based methodology to assemble the tetradecacyclic polyether skeleton. Convergent union of the GHI (5) and KLMN (6) rings, both of which were prepared from a common intermediate 7, and the subsequent ring closure of the J ring delivered the GHIJKLMN ring. The crucial coupling between the ABCD and FGHIJKLMN ring fragments (3 and 4, respectively) and stereoselective installation of the C17 hydroxyl group, followed by cyclization of the E ring gave rise to the tetradecacyclic polyether skeleton 2. Finally, incorporation of the 2-methyl-2 butenal side chain completed the total synthesis of gymnocin-A. The convergent nature of the synthesis, which employs three fragments of comparable complexity, is well-suited for preparation of various structural analogues of gymnocin-A to explore the structure-activity relationship. The results of preliminary structure activity relationship studies of several synthetic analogues are also provided. PMID- 15783213 TI - Tuning the orientation of an antigen by adsorption onto nanostriped templates. AB - We investigate the adsorption of a globular protein (P.69 pertactin, also known as antigen 69k) on protein-repellent hydrophilic substrates bearing regularly spaced hydrophobic nanostripes, for stripe widths comprised between 20 and 160 nm. Protein adsorption is shown to be remarkably well-controlled by the templating substrates, with a near-to-perfect reproduction of stripes by the protein monolayer down to 20 nm width, except for a 5-10 nm broadening. However, whereas the ellipsoidal protein forms a dense monolayer with random orientation of its long axis for large stripe widths, it adsorbs in a predominantly side-on (flat-on) orientation for stripe widths below 50 nm, due to the easier reorientation (interfacial relaxation) of the proteins adsorbed at the edges of the stripes, which experience a decreased lateral interaction. These results show that protein confinement in regions of a size similar to their dimensions can be used to tune their orientation, which may be of interest for applications in high density sensor devices. PMID- 15783216 TI - Interstellar chemistry: a strategy for detecting polycyclic aromatic hydrocarbons in space. AB - Polycyclic aromatic hydrocarbons (PAHs) have long been postulated as constituents of the interstellar gas and circumstellar disks. Observational infrared emission spectra have been plausibly interpreted in support of this hypothesis, but the small (or zero) dipole moments of planar, unsubstituted PAHs preclude their definitive radio astronomical identification. Polar PAHs, such as corannulene, thus represent important targets for radio astronomy because they offer the possibilities of confirming the existence of PAHs in space and revealing new insight into the chemistry of the interstellar medium. Toward this objective, the high-resolution rotational spectrum of corannulene has been obtained by Fourier transform microwave spectroscopy, and the dipole moment (2.07 D) of this exceptionally polar PAH has been measured by exploiting the Stark effect. PMID- 15783215 TI - Kinetics of 5alpha-cholestan-3beta-yl N-(2-naphthyl)carbamate/n-alkane organogel formation and its influence on the fibrillar networks. AB - The kinetics and mode of nucleation and growth of fibers by 5alpha-cholestan 3beta-yl N-(2-naphthyl)carbamate (CNC), a low-molecular-mass organogelator (LMOG), in n-octane and n-dodecane have been investigated as their sols were transformed isothermally to organogels. The kinetics has been followed in detail by circular dichroism, fluorescence, small-angle neutron scattering, and rheological methods. When treated according to Avrami theory, kinetic data from the four methods are self-consistent and describe a gelation process involving one-dimensional growth and "instantaneous nucleation". As expected from this growth model, polarized optical micrographs of the self-assembled fibrillar networks (SAFINs) show fibrous aggregates. However, their size and appearance change abruptly from spherulitic to rodlike as temperature is increased. This morphological change is attended by corresponding excursions in static and kinetic CD, fluorescence and rheological data. Furthermore, the rheological measurements reveal an unusual linear increase in viscoelastic moduli in the initial stages of self-assembly. Each of the methods employed becomes sensitive to changes of the system at different stages of the transformation from single molecules of the LMOG to their eventual SAFINs. This study also provides a methodology for investigating aggregation phenomena of some other self-assembling systems, including those of biological and physiological importance. PMID- 15783217 TI - Stereocontrolled synthesis and optical properties of all-cis poly(phenylene vinylenes) (PPVs): a method for direct patterning of PPVs. AB - Geometrically pure, all-cis poly(phenylene vinylenes) (PPVs) are synthesized by Suzuki-Miyaura-type polycondensation of 2,5-dioctyloxy-1,4-benzenediboronic acid with (Z,Z)-bis(2-bromoethenyl)benzenes, which are prepared by ruthenium-catalyzed (Z)-selective double hydrosilylation of diethynylbenzenes, followed by bromodesilylation of the resulting (Z,Z)-bis(2-silylethenyl)benzenes with N bromosuccinimide. The all-cis PPVs thus obtained undergo one-way photoisomerization to the corresponding trans-PPVs both in solution and in the solid. This phenomenon is applied to direct microscale patterning of PPVs onto a quartz substrate. PMID- 15783218 TI - Syntheses and structures of hypervalent pentacoordinate carbon and boron compounds bearing an anthracene skeleton--elucidation of hypervalent interaction based on X-ray analysis and DFT calculation. AB - Pentacoordinate and tetracoordinate carbon and boron compounds (27, 38, 50-52, 56 61) bearing an anthracene skeleton with two oxygen or nitrogen atoms at the 1,8 positions were synthesized by the use of four newly synthesized tridentate ligand precursors. Several carbon and boron compounds were characterized by X-ray crystallographic analysis, showing that compounds 27, 56-59 bearing an oxygen donating anthracene skeleton had a trigonal bipyramidal (TBP) pentacoordinate structure with relatively long apical distances (ca. 2.38-2.46 A). Despite the relatively long apical distances, DFT calculation of carbon species 27 and boron species 56 and experimental accurate X-ray electron density distribution analysis of 56 supported the existence of the apical hypervalent bond even though the nature of the hypervalent interaction between the central carbon (or boron) and the donating oxygen atom was relatively weak and ionic. On the other hand, X-ray analysis of compounds 50-52 bearing a nitrogen-donating anthracene skeleton showed unsymmetrical tetracoordinate carbon or boron atom with coordination by only one of the two nitrogen-donating groups. It is interesting to note that, with an oxygen-donating skeleton, the compound 61 having two chlorine atoms on the central boron atom showed a tetracoordinate structure, although the corresponding compound 60 with two fluorine atoms showed a pentacoordinate structure. The B-O distances (av 2.29 A) in 60 were relatively short in comparison with those (av 2.44 A) in 59 having two methoxy groups on the central boron atom, indicating that the B-O interaction became stronger due to the electron-withdrawing nature of the fluorine atoms. PMID- 15783219 TI - Investigation of the crystallization process in 2 nm CdSe quantum dots. AB - Investigation of the growth of CdSe nanocrystals ( approximately 160 atoms) to the uniquely stable size of 2 nm allows the monitoring of the crystallization process in semiconductor quantum dots. By using a combination of optical techniques, high-resolution transmission electron microscopy (HRTEM), and powder X-ray diffractometry (XRD), new phenomena were explored during the CdSe nanocrystal growth process, which involved significant morphological reconstruction and crystallization of the initially formed amorphous nanoparticles. During the crystallization, the absorption onset of the CdSe quantum dots blue shifted toward higher energies at 3 eV (414 nm), while the photoluminescence red shifted to lower energies. Furthermore, an apparent increasing Stokes shift was observed during the formation of small CdSe nanoparticles. On the other hand, the photoluminescence excitation spectra showed constant features over the reaction time. Additionally, results from HRTEM and XRD studies show that the CdSe nanoparticles were amorphous at early reaction stages and became better crystallized after longer reaction times, while the particle size remained the same during the crystallization process. These observations demonstrate the important role of the surface on the optical properties of small CdSe quantum dots and facilitated the spectroscopic monitoring of the crystallization process in quantum dots. PMID- 15783220 TI - Trimethylsilylated allyl complexes of nickel. The stabilized bis(pi-allyl)nickel complex [eta3-1,3-(SiMe3)2C3H3]2Ni and its mono(pi-allyl)NiX (X=Br, I) derivatives. AB - Reaction of 2 equiv of K[1,3-(SiMe3)2C3H3] with NiBr2(dme) in THF at -78 degrees C produces the orange pi-allyl complex [1,3-(SiMe3)2C3H3]2Ni (1). Unlike the pyrophoric (C3H5)2Ni, the trimethylsilylated derivative only slowly decomposes in air (from hours to days). Both eclipsed (1a) and staggered (1b) conformations are found in solution; the eclipsed form irreversibly converts to the thermodynamically more stable staggered conformation when heated above 85 degrees C. Single-crystal X-ray structures obtained for both 1a and 1b confirm that the allyl ligands are bound in a trihapto manner to the metals and that trimethylsilyl substituents are in syn, anti arrangements. Density functional theory calculations performed on the bis(allyl)nickel complexes indicate that the substituents exert little effect on the basic metal-ligand geometries. Trimethylphosphine is converted to tetramethyltetraphosphane, (MeP)4, on reaction with 1. In toluene, 3-bromo-1,3-bis(trimethylsilyl)propene reacts with (COD)2Ni to produce the dimeric purple complex {[1,3-(SiMe3)2C3H3]NiBr}2 (2a). Both NMR and X-ray crystallographic data establish that the allyl ligands are staggered and that the trimethylsilyl substituents are in a syn, syn conformation. NMR data indicate that the reaction of one equivalent of 1 with Br2 in benzene produces an analogous complex (2b) with the allyl ligand substituents in a syn, anti configuration. When 1 equiv of 1 is treated with I2 in hexanes, the dark red dimeric complex {[1,3-(SiMe3)2C3H3]NiI}2 (3) is formed. Its X-ray crystal structure demonstrates that both eclipsed (3a) and staggered (3b) allyl conformation are present. The trimethylsilyl groups on the allyl ligands are in syn, anti arrangements in the two forms. PMID- 15783221 TI - Binding and condensation of plasmid DNA onto functionalized carbon nanotubes: toward the construction of nanotube-based gene delivery vectors. AB - Carbon nanotubes (CNTs) constitute a class of nanomaterials that possess characteristics suitable for a variety of possible applications. Their compatibility with aqueous environments has been made possible by the chemical functionalization of their surface, allowing for exploration of their interactions with biological components including mammalian cells. Functionalized CNTs (f-CNTs) are being intensively explored in advanced biotechnological applications ranging from molecular biosensors to cellular growth substrates. We have been exploring the potential of f-CNTs as delivery vehicles of biologically active molecules in view of possible biomedical applications, including vaccination and gene delivery. Recently we reported the capability of ammonium functionalized single-walled CNTs to penetrate human and murine cells and facilitate the delivery of plasmid DNA leading to expression of marker genes. To optimize f-CNTs as gene delivery vehicles, it is essential to characterize their interactions with DNA. In the present report, we study the interactions of three types of f-CNTs, ammonium-functionalized single-walled and multiwalled carbon nanotubes (SWNT-NH3+; MWNT-NH3+), and lysine-functionalized single-walled carbon nanotubes (SWNT-Lys-NH3+), with plasmid DNA. Nanotube-DNA complexes were analyzed by scanning electron microscopy, surface plasmon resonance, PicoGreen dye exclusion, and agarose gel shift assay. The results indicate that all three types of cationic carbon nanotubes are able to condense DNA to varying degrees, indicating that both nanotube surface area and charge density are critical parameters that determine the interaction and electrostatic complex formation between f-CNTs with DNA. All three different f-CNT types in this study exhibited upregulation of marker gene expression over naked DNA using a mammalian (human) cell line. Differences in the levels of gene expression were correlated with the structural and biophysical data obtained for the f-CNT:DNA complexes to suggest that large surface area leading to very efficient DNA condensation is not necessary for effective gene transfer. However, it will require further investigation to determine whether the degree of binding and tight association between DNA and nanotubes is a desirable trait to increase gene expression efficiency in vitro or in vivo. This study constitutes the first thorough investigation into the physicochemical interactions between cationic functionalized carbon nanotubes and DNA toward construction of carbon nanotube based gene transfer vector systems. PMID- 15783222 TI - Molecular rotor of Cs2([18]crown-6)3 in the solid state coupled with the magnetism of [Ni(dmit)2]. AB - Nanoscale molecular rotors that can be driven in the solid state have been realized in Cs2([18]crown-6)3[Ni(dmit)2]2 crystals. To provide interactions between the molecular motion of the rotor and the electronic system, [Ni(dmit)2]- ions, which bear one S=1/2 spin on each molecule, were introduced into the crystal. Rotation of the [18]crown-6 molecules within a Cs2([18]crown-6)3 supramolecule above 220 K was confirmed using X-ray diffraction, NMR, and specific heat measurements. Strong correlations were observed between the magnetic behavior of the [Ni(dmit)2]- ions and molecular rotation. Furthermore, braking of the molecular rotation within the crystal was achieved by the application of hydrostatic pressure. PMID- 15783223 TI - The electronic structure of the isoelectronic, square-planar complexes [FeII(L)2]2- and [CoIII(L Bu)2]- (L2- and (L Bu)2-=benzene-1,2-dithiolates): an experimental and density functional theoretical study. AB - The electronic structures of two formally isoelectronic transition-metal dithiolato complexes [Fe(L)2]2- (1) and [Co(L Bu)2]1- (2) both possessing a spin triplet ground state (St=1) have been investigated by various spectroscopic and density functional methods; H2L Bu represents the pro-ligand 3,5-di-tert butylbenzene-1,2-dithiol and H2L is the corresponding unsubstituted benzene-1,2 dithiol. An axial zero-field splitting (D) of +32 cm(-1) for 2 has been measured independently by SQUID magnetometry, far-infrared absorption, and variable temperature and variable-field (VTVH) magnetic circular dichroism spectroscopies. A similar D value of +28 cm(-1) is obtained for 1 on the basis of VTVH SQUID measurements. The absorption spectra of 1 and 2 are found, however, to be very different. Complex 1 is light yellow in color with no intense transition in the visible region, whereas 2 is deep blue. DFT calculations establish that the electronic structures of the [Fe(L)2](2-) and [Co(L)2]1- anions are very different and explain the observed differences in their absorption spectra. On the basis of these spectroscopic and theoretical analyses, 1 is best described as containing an intermediate spin FeII ion, whereas for the corresponding cobalt complex, oxidation states describing a d6 (CoIII) or d7 (CoII) electron configuration cannot be unambiguously assigned. The physical origin of the large zero-field splitting in both 1 and 2 is found to be due to the presence of low energy spin-conserved d-d excitations which lead to a large Dzz through efficient spin-orbit coupling. Differential covalency effects appear to be of limited importance for this property. PMID- 15783224 TI - Quantitative determination of the chemical composition of silica-poly(norbornene) nanocomposites. AB - Nanoparticle hybrid materials consisting of a silica core surrounded by a poly(norbornene) brush have been prepared by ring opening metathesis polymerization (ROMP). A quantitative determination of each stage of composite formation has been accomplished, including a determination of the density of surface-bound functional groups, catalyst molecules, and polymer chains. This analysis has enabled the determination of the reaction efficiency between the catalyst and the surface-bound functional groups as well as the determination of the fraction of metal-mediating species that initiate a polymer chain. Control of the chain density was demonstrated by two methods: the use of controlled reaction times between the catalyst and the surface, and the variation of the surface functional group density. Polymer chain densities resulting from composites prepared with different tether structures will also be reported. The resulting brush densities were found to span a wide range, including those previously reported for polymer layers formed by adsorption, grafting of preformed polymer chains, and surface-initiated polymerization (SIP). PMID- 15783225 TI - Is it homogeneous or heterogeneous catalysis? Compelling evidence for both types of catalysts derived from [Rh(eta5-C5Me5)Cl2]2 as a function of temperature and hydrogen pressure. AB - Addressed herein is the 20+ year-old question of whether the true benzene and cyclohexene hydrogenation catalysts derived from the organometallic precursor [Rh(eta5-C5Me5)Cl2]2, 1, are homogeneous or heterogeneous. The methodology employed is that developed earlier (Lin, Y.; Finke, R. G. Inorg Chem. 1994, 33, 4891, "A More General Approach to Distinguishing Homogeneous from Heterogeneous Catalysis..."). The kinetic evidence especially, but also the metal product (nanoclusters plus bulk metal), Hg0 poisoning and other experiments, provide compelling evidence that Rh0 nanoclusters are the true benzene hydrogenation heterogeneous catalyst derived from [Rh(eta5-C5Me5)Cl2]2, 1, at the required more vigorous conditions of 50-100 degrees C and 50 atm H2. However, the same methods reveal that the cyclohexene hydrogenation catalyst derived from 1 at the milder conditions of 22 degrees C and 3.7 atm H2 is a nonnanocluster, homogeneous catalyst, most likely the previously identified complex, [Rh(eta5 C5Me5)(H)2(solvent)] (Gill, D. S.; White, C.; Maitlis, P. M J. C. S. Dalton Trans. 1978, 617). In short, the present results solve the two-decade-old problem of identifying the true benzene and cyclohexene hydrogenation catalysts derived from [Rh(eta5-C5Me5)Cl2]2. Perhaps most significant is the demonstration that the methodology employed has the ability to identify both heterogeneous and homogeneous catalysts from the same catalyst precursor. PMID- 15783226 TI - Theoretical study of the truncated hemoglobin HbN: exploring the molecular basis of the NO detoxification mechanism. AB - Mycobacterium tuberculosis is the causative agent of human tuberculosis. The nitric oxide reaction with oxy-truncated hemoglobin N (trHbN) has been proposed to be responsible for the resistance mechanism by which this microorganism can evade the toxic effects of NO. In this work, we explore the molecular basis of the NO detoxification mechanism using a combination of classical and hybrid quantum-classical (QM-MM) simulation techniques. We have investigated the structural flexibility of the protein, the ligand affinity properties, and the nitric oxide reaction with coordinated O2. The analysis of the classical MD trajectory allowed us to identify Phe62 as the gate of the main channel for ligand diffusion to the active site. Moreover, the opening of the channel stems from the interplay between collective backbone motions and local rearrangements in the side chains of the residues that form the bottleneck of the tunnel. Even though the protein environment is not found to make a significant contribution to the heme moiety catalyzed reaction, the binding site influences the physiological function of the enzyme at three different levels. First, by isolating the intermediates formed in the reaction, it prevents nondesired reactions from proceeding. Second, it modulates the ligand (O2, NO) affinity of the protein, which can be ascribed to both distal and proximal effects. Finally, the stabilization of the Tyr33-Gln58 pair upon O2 binding might alter the essential dynamics of the protein, leading in turn to a mechanism for ligand-induced regulation. PMID- 15783227 TI - Water-promoted hydrolysis of a highly twisted amide: rate acceleration caused by the twist of the amide bond. AB - The water-promoted hydrolysis of a highly twisted amide is studied using density functional theory in conjunction with a continuum dielectric method to introduce bulk solvent effects. The aim of these studies is to reveal how the twisting of the C-N bond affects the neutral hydrolysis of amides. To do so, both concerted and stepwise mechanisms are studied and the results compared to the ones from the hydrolysis of an undistorted amide used as reference. In addition, an extra explicit water molecule that assists in the required proton-transfer processes is taken into account. Our results predict important rate accelerations of the neutral hydrolysis of amides when the C-N bond is highly twisted, the corresponding barrier relaxation depending on the specific reaction pathway and transition state involved. Moreover, our calculations strongly suggest a change in reaction mechanism with degree of amide bond twist, and clearly point to a concerted mechanism at neutral pH for the hydrolysis of highly twisted amides. In addition, the twisting of the amide bond also provokes a higher dependence on an auxiliary water molecule for the concerted mechanism, due to the orthogonality of the lone pair of the nitrogen and the carbonyl pi orbital. There is a direct implication of these findings for biological catalytic mechanism of peptide cleavage reactions that undergoes ground-state destabilization of the peptide. PMID- 15783228 TI - Mechanisms of antibiotic resistance: QM/MM modeling of the acylation reaction of a class A beta-lactamase with benzylpenicillin. AB - Understanding the mechanisms by which beta-lactamases destroy beta-lactam antibiotics is potentially vital in developing effective therapies to overcome bacterial antibiotic resistance. Class A beta-lactamases are the most important and common type of these enzymes. A key process in the reaction mechanism of class A beta-lactamases is the acylation of the active site serine by the antibiotic. We have modeled the complete mechanism of acylation with benzylpenicillin, using a combined quantum mechanical and molecular mechanical (QM/MM) method (B3LYP/6-31G+(d)//AM1-CHARMM22). All active site residues directly involved in the reaction, and the substrate, were treated at the QM level, with reaction energies calculated at the hybrid density functional (B3LYP/6-31+Gd) level. Structures and interactions with the protein were modeled by the AM1 CHARMM22 QM/MM approach. Alternative reaction coordinates and mechanisms have been tested by calculating a number of potential energy surfaces for each step of the acylation mechanism. The results support a mechanism in which Glu166 acts as the general base. Glu166 deprotonates an intervening conserved water molecule, which in turn activates Ser70 for nucleophilic attack on the antibiotic. This formation of the tetrahedral intermediate is calculated to have the highest barrier of the chemical steps in acylation. Subsequently, the acylenzyme is formed with Ser130 as the proton donor to the antibiotic thiazolidine ring, and Lys73 as a proton shuttle residue. The presented mechanism is both structurally and energetically consistent with experimental data. The QM/MM energy barrier (B3LYP/ 6-31G+(d)//AM1-CHARMM22) for the enzymatic reaction of 9 kcal mol(-1) is consistent with the experimental activation energy of about 12 kcal mol(-1). The effects of essential catalytic residues have been investigated by decomposition analysis. The results demonstrate the importance of the "oxyanion hole" in stabilizing the transition state and the tetrahedral intermediate. In addition, Asn132 and a number of charged residues in the active site have been identified as being central to the stabilizing effect of the enzyme. These results will be potentially useful in the development of stable beta-lactam antibiotics and for the design of new inhibitors. PMID- 15783229 TI - Chemical shift correlations in disordered solids. AB - We show how two-dimensional chemical shift conditional probability distributions can be extracted from experimental NMR correlation spectra of disordered solids. We show that transverse dephasing times are of central importance in determining the resolution (and sensitivity) of these probability distributions. These conditional probability distributions provide a new source of structural information characteristic of disordered solids, which is much more sensitive to structure than the individual-atom chemical shift distributions. The structural information contained in these distributions is clearly a potentially extremely rich source for understanding disorder. This is illustrated with examples of a phosphorus-containing organic compound and with a sample of disordered cellulose where the different structural allomorphs present are identified from the distributions. PMID- 15783230 TI - Determination of peptide oligomerization in lipid bilayers using 19F spin diffusion NMR. AB - Aggregation or oligomerization is important for the function of many membrane peptides such as ion channels and antimicrobial peptides. However, direct proof of aggregation and the determination of the number of molecules in the aggregate have been difficult due to the lack of suitable high-resolution methods for membrane peptides. We propose a 19F spin diffusion magic-angle-spinning NMR technique to determine the oligomeric state of peptides bound to the lipid bilayer. Magnetization transfer between chemically equivalent but orientationally different 19F spins on different molecules reduces the 19F magnetization in an exchange experiment. At long mixing times, the equilibrium 19F magnetization is 1/M, where M is the number of orientationally different molecules in the aggregate. The use of the 19F spin increases the homonuclear dipolar coupling and thus the distance reach. We demonstrate this technique on crystalline model compounds with known numbers of molecules in the asymmetric unit cell, and show that 19F spin diffusion is more efficient than that of 13C by a factor of approximately 500. Application to a beta-hairpin antimicrobial peptide, protegrin 1, shows that the peptide is almost completely dimerized in POPC bilayers at a concentration of 7.4 mol %. Decreasing the peptide concentration reduced the dimer fraction. Using a monomer-dimer equilibrium model, we estimate the DeltaG for dimer formation to be -10.2 +/- 2.3 kJ/mol. This is in good agreement with the previously measured free energy reduction for partitioning and aggregating beta-sheet peptides into phospholipid membranes. This 19F spin diffusion technique opens the possibility of determining the oligomeric structures of membrane peptides. PMID- 15783231 TI - Rapid detection of an anthrax biomarker by surface-enhanced Raman spectroscopy. AB - A rapid detection protocol suitable for use by first-responders to detect anthrax spores using a low-cost, battery-powered, portable Raman spectrometer has been developed. Bacillus subtilis spores, harmless simulants for Bacillus anthracis, were studied using surface-enhanced Raman spectroscopy (SERS) on silver film over nanosphere (AgFON) substrates. Calcium dipicolinate (CaDPA), a biomarker for bacillus spores, was efficiently extracted by sonication in nitric acid and rapidly detected by SERS. AgFON surfaces optimized for 750 nm laser excitation have been fabricated and characterized by UV-vis diffuse reflectance spectroscopy and SERS. The SERS signal from extracted CaDPA was measured over the spore concentration range of 10(-14)-10(-12) M to determine the saturation binding capacity of the AgFON surface and to calculate the adsorption constant (Kspore=1.7 x 10(13) M(-1)). At present, an 11 min procedure is capable of achieving a limit of detection (LOD) of approximately 2.6 x 10(3) spores, below the anthrax infectious dose of 10(4) spores. The data presented herein also demonstrate that the shelf life of prefabricated AgFON substrates can be as long as 40 days prior to use. Finally, these sensing capabilities have been successfully transitioned from a laboratory spectrometer to a field-portable instrument. Using this technology, 10(4) bacillus spores were detected with a 5 s data acquisition period on a 1 month old AgFON substrate. The speed and sensitivity of this SERS sensor indicate that this technology can be used as a viable option for the field analysis of potentially harmful environmental samples. PMID- 15783232 TI - Viscosity of the aqueous liquid/vapor interfacial region: 2D electrochemical measurements with a piperidine nitroxy radical probe. AB - Surface partitioning of 2,2,6,6-tetramethyl-1-piperidynyloxy radical (Tempo) to the air/water interface follows a Langmuir isotherm. The partition constant was obtained by the surface tension measurements in the concentration range of 1.0 x 10(-4)-2.4 x 10(-3) M yielding K = 640 +/- 99 M(-1). The lateral mobility of Tempo at the air/water interface was measured electrochemically in the surface concentration range of 2.0 x 10(-11)-1.4 x 10(-10) mol/cm2, corresponding to ca. 7.3-50% full monolayer coverage. The measurements employed cyclic voltammetry with line microelectrodes touching the air/water interface. The Tempo lateral diffusion constant of (1.5 +/- 0.7) x 10(-4) cm2/s is independent of surface concentration below 4.0 x 10(-11) mol/cm2. The extent of Tempo water interactions was assessed by the electronic structure calculations. These calculations showed that, at most, two water molecules can hydrogen bond with the oxygen atom of the nitroxyl group of Tempo, and that a single water molecule forms a hydrogen bond that is ca. 30% stronger than the H2O-H2O hydrogen bond. These calculations led to a postulate that Tempo diffuses along the interface largely unimmersed, and that it is coupled to the interfacial water via hydrogen bonding with H2O. In view of this postulate, the viscosity of the aqueous liquid/vapor interfacial region obtained by interpreting the Tempo diffusion constant in the low concentration region is as much as 4 times smaller than that of bulk liquid water. PMID- 15783233 TI - Separation of single-walled carbon nanotubes on silica gel. Materials morphology and Raman excitation wavelength affect data interpretation. AB - In this report, procedures are discussed for the enrichment of single-walled carbon nanotube (SWNT) types by simple filtration of the functionalized SWNTs through silica gel. This separation uses nanotube sidewall functionalization employing two different strategies. In the first approach, a crude mixture of metallic and semiconducting SWNTs was heavily functionalized with 4-tert butylphenyl addends to impart solubility to the entire sample of SWNTs. Two major polarity fractions were rapidly filtered through silica gel, with the solvent being removed in vacuo, heated to 700 degrees C to remove the addends, and analyzed spectroscopically. The second approach uses two different aryldiazonium salts (one with a polar grafting group and one nonpolar), appended selectively onto the different SWNTs by means of titration and monitoring by UV analysis throughout the functionalization process. The different addends accentuate the polarity differences between the band-gap-based types permitting their partial separation on silica gel. Thermal treatment regenerated pristine SWNTs in enriched fractions. The processed samples were analyzed and characterized by Raman spectroscopy. A controlled functionalization method using 4-fluorophenyl and 4-iodophenyl addends was performed, and XPS analyses yielded data on the degree of functionalization needed to affect the van Hove singularities in the UV/vis/NIR spectra. Finally, we demonstrate that relative peak intensity changes in Raman spectra can be caused by morphological changes in SWNT bundling based on differing flocculation or deposition methods. Therefore a misleading impression of separations can result, underscoring the care needed in assessing efficacies in SWNT enrichment and the prerequisite use of multiple excitation wavelengths and similar flocculation or deposition methods in comparative analyses. PMID- 15783234 TI - Regarding the mechanism of olefin metathesis with sol-gel-supported Ru-based complexes bearing a bidentate carbene ligand. Spectroscopic evidence for return of the propagating Ru carbene. AB - Two isotopically and structurally labeled Ru-based carbenes (2-d4 and 13) have been prepared and attached to the surface of monolithic sol-gel glass. The resulting glass-supported complexes (18-dn and 19) exhibit significant catalytic activity in promoting olefin metathesis reactions and provide products of high purity. Through analysis of the derivatized glass pellets used in a sequence of catalytic ring-closing metathesis reactions mediated by various supported Ru carbenes, it is demonstrated that free Ru carbene intermediates in solution can be scavenged by support-bound styrene ether ligands prior to the onset of competing transition metal decomposition. The observations detailed herein provide rigorous evidence that the initially proposed release/return mechanism is, at least partially, operative. The present investigations shed light on a critical aspect of the mechanism of an important class of Ru-based metathesis complexes (those bearing a bidentate styrene ether ligand). PMID- 15783235 TI - Nature of the catalytically labile oxygen at the active site of xanthine oxidase. AB - In this paper we report the results of molybdenum K-edge X-ray absorption studies performed on the oxidized active site of xanthine oxidase at pH 6 and 10. These results indicate that the active site possesses one terminal oxygen ligand (Mo=O), two thiolate ligands (Mo-S), one terminal sulfido ligand (Mo=S), and one Mo-OH moiety. EXAFS analysis demonstrates that the Mo-OH bond shortens from 1.97 A at pH 6 to 1.75 A at pH 10, which is consistent with the generation of a Mo-O- moiety. This study provides convincing structural evidence that the catalytic oxygen donor at the oxidized active site of xanthine oxidase is Mo-OH rather than the Mo-OH2 ligation previously suggested by X-ray crystallography. These results support a mechanism initiated by base-assisted nucleophilic attack of the substrate by Mo-OH. PMID- 15783236 TI - Size-dependent phase transformation kinetics in nanocrystalline ZnS. AB - Nanocrystalline ZnS was coarsened under hydrothermal conditions to investigate the effect of particle size on phase transformation kinetics. Although bulk wurtzite is metastable relative to sphalerite below 1020 degrees C at low pressure, sphalerite transforms to wurtzite at 225 degrees C in the hydrothermal experiments. This indicates that nanocrystalline wurtzite is stable at low temperature. High-resolution transmission electron microscope data indicate there are no pure wurtzite particles in the coarsened samples and that wurtzite only grows on the surface of coarsened sphalerite particles. Crystal growth of wurtzite stops when the diameter of the sphalerite-wurtzite interface reaches approximately 22 nm. We infer that crystal growth of wurtzite is kinetically controlled by the radius of the sphalerite-wurtzite interface. A new phase transformation kinetic model based on collective movement of atoms across the interface is developed to explain the experimental results. PMID- 15783237 TI - Substituent effects on the edge-to-face aromatic interactions. AB - The edge-to-face interactions for either axially or facially substituted benzenes are investigated by using ab initio calculations. The predicted maximum energy difference between substituted and unsubstituted systems is approximately 0.7 kcal/mol (approximately 1.2 kcal/mol if substituents are on both axially and facially substituted positions). In the case of axially substituted aromatic systems, the electron density at the para position is an important stabilizing factor, and thus the stabilization/destabilization by substitution is highly correlated to the electrostatic energy. This results in its subsequent correlation with the polarization and charge transfer. Thus, the stabilization/destabilization by substitution is represented by the sum of electrostatic energy and induction energy. On the other hand, the facially substituted aromatic system depends on not only the electron-donating ability responsible for the electrostatic energy but also the dispersion interaction and exchange repulsion. Although the dispersion energy is the most dominating interaction in both axial and facial substitutions, it is almost canceled by the exchange repulsion in the axial substitution, whereas in the facial substitution, together with the exchange repulsion it augments the electrostatic energy. The systems with electron-accepting substituents (NO2, CN, Br, Cl, F) favor the axial substituent conformation, while those with electron-donating substituents (NH2, CH3, OH) favor the facial substituent conformation. The interactions for the T shape complex systems of an aromatic ring with other counterpart such as H2, H2O, HCl, and HF are also studied. PMID- 15783238 TI - The WHO 'Roll Back Malaria Project': planning for adverse event monitoring in Africa. AB - Artemisinin combination therapies (ACTs) have been recommended for the treatment of malaria in countries where there is widespread resistance to commonly used antimalarial drugs. Several sub-Saharan African countries are, therefore, in the process of introducing ACTs in their malaria drug policies. However, there is limited information about the safety of ACTs outside South East Asia, where their use has been well documented. As with all other new medicinal compounds, the monitoring of a drug's safety or 'pharmacovigilance' is important, especially in areas where co-morbid conditions, such as HIV/AIDS, malnutrition and tuberculosis, are common. Because in most malaria endemic countries, particularly Africa, there are no pharmacovigilance programmes in place, it has been suggested that the introduction of ACTs offers an opportunity for these countries to put drug safety monitoring systems in place. Backed by the WHO Roll Back Malaria department and other international cooperating partners, five African countries, which are in the process of introducing ACTs (Burundi, Democratic Republic of the Congo, Mozambique, Zambia and Zanzibar), have drawn up action plans to introduce pharmacovigilance in their health sector. It is planned that once the safety monitoring of antimalarials has been established, these activities can then be extended to cover medicinal compounds used in other public health programmes, such as HIV/AIDS, tuberculosis and the immunisation programmes. This article looks at the rationale for pharmacovigilance, the process of setting up monitoring centres and the challenges of implementing the project in the region. PMID- 15783239 TI - NSAID-induced peptic ulcers and Helicobacter pylori infection: implications for patient management. AB - The conflicting data about the influence of Helicobacter pylori infection on the ulcer risk in patients receiving NSAIDs can be accounted for by the heterogeneity of study designs and the diversified host response to H. pylori. Factors that will affect the outcome include the choice of H. pylori diagnostic tests, previous ulcer complications, concurrent use of acid suppressants, NSAID-naive versus long-term users, low-dose aspirin (acetylsalicylic acid) versus non aspirin NSAIDs and whether the result was derived from a pre-specified endpoint or post hoc subgroup analysis. Current evidence suggests that H. pylori eradication reduces the ulcer risk for patients who are about to start receiving NSAIDs but not for those who are already on long-term NSAID therapy. Since treatment with a proton pump inhibitor (PPI) worsens H. pylori-associated corpus gastritis, H. pylori should be tested for, and eradicated if present, before starting long-term prophylaxis with PPIs. Patients with H. pylori infection and a history of ulcer complications who require NSAIDs should receive concomitant PPIs or misoprostol after curing the infection. Among patients receiving low-dose aspirin, who have H. pylori infection and previous ulcer complications, long-term treatment with a PPI further reduces the risk of complicated ulcers if H. pylori eradication fails or if patients use concomitant non-aspirin NSAIDs. Current data on the gastric safety of COX-2 selective NSAIDs in H. pylori-infected patients are conflicting. Limited data suggest that the gastroduodenal sparing effect of rofecoxib is negated by H. pylori infection in patients who have had prior upper gastrointestinal events. In light of potential cardiovascular risk with COX-2 selective NSAIDs, it is important to weigh the potential adverse effects against the benefits for an individual patient. PMID- 15783241 TI - Vitex agnus castus: a systematic review of adverse events. AB - Vitex agnus castus L. (VAC) [Verbenaceae] is a deciduous shrub that is native to Mediterranean Europe and Central Asia. Traditionally, VAC fruit extract has been used in the treatment of many female conditions, including menstrual disorders (amenorrhoea, dysmenorrhoea), premenstrual syndrome (PMS), corpus luteum insufficiency, hyperprolactinaemia, infertility, acne, menopause and disrupted lactation. The German Commission E has approved the use of VAC for irregularities of the menstrual cycle, premenstrual disturbances and mastodynia. Clinical reviews are available for the efficacy of VAC in PMS, cycle disorders, hyperprolactinaemia and mastalgia, but so far no systematic review has been published on adverse events or drug interactions associated with VAC. Therefore, this review was conducted to evaluate all the available human safety data of VAC monopreparations. Literature searches were conducted in six electronic databases, in references lists of all identified papers and in departmental files. Data from spontaneous reporting schemes of the WHO and national drug safety bodies were also included. Twelve manufacturers of VAC-containing preparations and five herbalist organisations were contacted for additional information. No language restrictions were imposed. Combination preparations including VAC or homeopathic preparations of VAC were excluded. Data extraction of key data from all articles reporting adverse events or interactions was performed independently by at least two reviewers, regardless of study design. Data from clinical trials, postmarketing surveillance studies, surveys, spontaneous reporting schemes, manufacturers and herbalist organisations indicate that the adverse events following VAC treatment are mild and reversible. The most frequent adverse events are nausea, headache, gastrointestinal disturbances, menstrual disorders, acne, pruritus and erythematous rash. No drug interactions were reported. Use of VAC should be avoided during pregnancy or lactation. Theoretically, VAC might also interfere with dopaminergic antagonists. Although further rigorous studies are needed to assess the safety of VAC, the data available seem to indicate that VAC is a safe herbal medicine. PMID- 15783240 TI - Benefit-risk assessment of zaleplon in the treatment of insomnia. AB - Insomnia is a heterogeneous, highly prevalent condition that is associated with a high level of psychiatric, physical, social and economic morbidity. The treatment of insomnia involves pharmacological and non-pharmacological interventions. The mainstay of pharmacological treatment of insomnia has been the benzodiazepines, the introduction of which represented a significant improvement over the barbiturates and chloral hydrate. Although benzodiazepines have been shown to be efficacious in treating insomnia, they have also been associated with a number of adverse effects including tolerance, dependence, withdrawal and abuse potential, impairment in daytime cognitive and psychomotor performance (including an increased risk of accidents and falls), adverse effects on respiration and the disruption of normal sleep architecture with reduction in both slow wave sleep and rapid eye movement. In the last decade, the treatment of insomnia has been supplemented by the introduction of a number of non-benzodiazepine hypnotics including zolpidem, zopiclone and, most recently, zaleplon. Zaleplon possesses a unique pharmacological profile, with an ultra-short half-life of about 1 hour, and selective binding to the BZ1(omega1) receptor subtypes of the GABA(A) receptor. This unique pharmacological profile predicts a number of pharmacodynamic properties that account for a unique benefit-risk profile. Consistent with these predictions, zaleplon has been shown in a number of studies to be efficacious in promoting sleep initiation, but less so in promoting sleep maintenance. The adverse effects associated with zaleplon have been shown to be more rapidly resolved and/or lesser in magnitude than those associated with benzodiazepines (including triazolam) and the longer acting non-benzodiazepine hypnotics (zolpidem and zopiclone). This improved risk profile includes: the effects of zaleplon on psychomotor and cognitive performance; tolerance, withdrawal and rebound; respiratory depression; sleep architecture; and other treatment-emergent adverse effects. The unique benefit-risk profile of this agent may be particularly suitable for certain patients with insomnia and provides yet another option in the management of this impairing condition. PMID- 15783242 TI - Safety profile of different low-molecular weight heparins used at therapeutic dose. AB - Low-molecular weight heparins (LMWHs) have been shown to be as safe and effective as unfractionated heparin (UFH) for the treatment of acute venous thrombosis and non-life-threatening pulmonary embolism. Different reports have shown that LMWHs may also be used to treat patients with unstable angina or non-Q-wave infarction. The safety of LMWHs used at therapeutic dose has been widely studied in pivotal clinical trials and analysed in several meta-analyses. However, despite the wide development and use of LMWHs, several issues regarding the safety and optimal use of LMWHs remain unanswered. The main adverse effect of LMWHs is bleeding and it is uncertain whether a weight-adjusted dosage regimen without laboratory monitoring can be used in patients with a high risk of bleeding, such as patients with renal failure, elderly patients, obese patients or pregnant women. These patients are usually excluded from clinical trials and only a few studies, not sufficiently powered to estimate efficacy and safety, have been carried out in these special populations. Most of the available data comes from pharmacokinetic or population pharmacodynamic studies or clinical reports. Results in patients with renal impairment who are not undergoing haemodialysis suggest that a reduction in calculated creatinine clearance levels is associated with an increased risk of accumulation of anti-Xa activity, the extent of which differs depending on the individual LMWH and the extent to which the compound is cleared by the kidney. The limited data available regarding the use of therapeutic doses of LMWHs in obese patients suggest that there is no need to cap the dose at a maximal allowable dose. Long-term (3-month) treatment with LMWHs appears to be as effective and safe as oral anticoagulant therapy for the treatment of venous thromboembolism. It appears that each LMWH is a distinct compound with unique pharmacokinetic and pharmacodynamic profiles. Until more data are available regarding these special populations, periodic monitoring of anti-Xa activity levels may be recommended to detect accumulation and/or an overdose and minimise the bleeding risk. The non-haemorrhagic adverse effects of the LMWHs include heparin-induced thrombocytopenia (HIT) and osteoporosis. The incidence of HIT appears to be lower with LMWHs than with UFH; there is currently not enough data to compare the frequency of HIT between the various LMWHs. LMWHs also appear to carry a lower risk of causing osteoporosis than UFH. In conclusion, studies that include special population patients are required to make conclusive recommendations concerning the safety and monitoring of the different LMWHs. PMID- 15783244 TI - Diabesity: are weight loss medications effective? AB - Weight reduction has been shown to improve glycemic control and cardiovascular risk factors associated with insulin resistance in obese individuals with type 2 diabetes mellitus. Therapeutic options for these patients include promoting weight loss (non-pharmacologic and pharmacologic treatment) and improving glycemic control, as well as treating common associated risk factors such as arterial hypertension and dyslipidemias. This article provides an overview of anti-obesity drugs used in the treatment of obese individuals with type 2 diabetes. The most widely investigated drugs, sibutramine and orlistat, result in modest, clinically worthwhile weight loss, with demonstrable improvements in many co-morbidities, among them, type 2 diabetes. Clinical trials with these anti obesity medications in cohorts of obese diabetic patients have been reviewed as well as cathecolaminergic agents (diethylpropion [amfepramone], fenproporex, mazindol, ephedrine-caffeine combination), serotoninergic drugs (fenfluramine, dexfenfluramine, fluoxetine), and other drugs that have some action on weight loss (the antidiabetic agent metformin, anti-epileptic agents topiramate and zonisamide, and the antidepressive bupropion [amfebutamone]). These trials show variable benefits in terms of effects on glucose profiles. PMID- 15783245 TI - Cardiovascular disease morbidity and mortality in patients with type 1 diabetes mellitus : management strategies. AB - There is an increased risk of cardiovascular disease (CVD) mortality and morbidity in patients with type 1 diabetes mellitus compared with the general population as shown by epidemiologic studies measuring cardiovascular endpoints, as well as by autopsy, angiographic, and coronary calcification studies. Most of the excess CVD risk associated with type 1 diabetes is concentrated in the subset of approximately 35% of patients who develop diabetic nephropathy (after 20 years of diabetes duration), who also typically have dyslipidemias, elevated blood pressure, and hyperglycemia, factors contributing to CVD. For reasons that remain speculative, the relative risks from CVD are higher in women than in men with type 1 diabetes compared with the general population, which effectively eliminates the gender differences in CVD. As in the general population and in patients with type 2 diabetes, education and lifestyle changes, interventions to reduce hyperglycemia, blood pressure, micro-albuminuria, lipid control, and the use of aspirin are important management areas in order to reduce the increased risk of CVD. Whether management with aspirin and statins should be started in type 1 diabetic patients at a younger age or at a lower risk score than in the general population is still under investigation. There is a need for a better understanding of the pathophysiology of vascular complications in type 1 diabetes, more specific risk engines in type 1 diabetes, and accurate estimations of the absolute and relative risk for CVD in order to improve management of CVD in these high-risk patients. PMID- 15783243 TI - Hepatic findings in long-term clinical trials of ximelagatran. AB - OBJECTIVE: In clinical trials, the efficacy and safety of the oral direct thrombin inhibitor ximelagatran have been evaluated in the prevention or treatment of thromboembolic conditions known to have high morbidity and mortality. In these studies, raised aminotransferase levels were observed during long-term use (>35 days). The aim of this analysis is to review the data regarding these hepatic findings in the long-term trials of ximelagatran. PATIENTS AND METHODS: The prospective analysis included 6948 patients randomised to ximelagatran and 6230 patients randomised to comparator (warfarin, low molecular weight heparin followed by warfarin or placebo). Of these, 6931 patients received ximelagatran for a mean of 357 days and 6216 patients received comparator for a mean of 389 days. An algorithm was developed for frequent testing of hepatic enzyme levels. A panel of four hepatologists analysed all cases of potential concern with regard to causal relation to ximelagatran treatment using an established evaluation tool (Roussel Uclaf Causality Assessment Method [RUCAM]). RESULTS: An elevated alanine aminotransferase (ALT) level of >3 x the upper limit of normal (ULN) was found in 7.9% of patients in the ximelagatran group versus 1.2% in the comparator group. The increase in ALT level occurred 1-6 months after initiation of therapy and data were available to confirm recovery of the ALT level to <2 x ULN in 96% of patients, whether they continued to receive ximelagatran or not. There was some variability in the incidence of ALT level elevation between indications, those with simultaneous acute illnesses (acute myocardial infarction or venous thromboembolism) having higher incidences. Combined elevations of ALT level of >3 x ULN and total bilirubin level of >2 x ULN (within 1 month of the ALT elevation), regardless of aetiology, were infrequent, occurring in 37 patients (0.5%) treated with ximelagatran, of whom one sustained a severe hepatic illness that appeared to be resolving when the patient died from a gastrointestinal haemorrhage. No death was observed directly related to hepatic failure caused by ximelagatran. CONCLUSION: Treatment with ximelagatran has been associated with mainly asymptomatic elevation of ALT levels in a mean of 7.9% of patients in the long-term clinical trial programme and nearly all of the cases occurred within the first 6 months of therapy. Rare symptomatic cases have been observed. An algorithm has been developed for testing ALT to ensure appropriate management of patients with elevated ALT levels. Regular ALT testing should allow the clinical benefits of ximelagatran to reach the widest population of patients while minimising the risk of hepatic adverse effects. PMID- 15783246 TI - Pharmacologic management of Cushing syndrome : new targets for therapy. AB - The successful treatment of Cushing syndrome depends on specific therapy directed against the etiology of hypercortisolism. In addition to surgical procedures, various drugs have been employed in the management of this difficult disease. Compounds with neuromodulatory properties have been effective in only a limited number of cases of hypothalamic-pituitary-dependent Cushing disease, the most common form of Cushing syndrome. These agents include serotonin antagonists (cyproheptadine, ketanserin, ritanserin), dopamine agonists (bromocriptine, cabergoline), GABA agonists (valproic acid [sodium valproate]), and somatostatin analogs (octreotide). Interesting new avenues at the pituitary level involve the potential use of thiazolidinedione compounds, such as rosiglitazone, and of retinoic acid, which are ligands of different nuclear hormone receptors involved in hypothalamic-pituitary regulation. The most exciting news, however, in the pharmacologic approach to Cushing syndrome refers to the adrenal corticotropin (adrenocorticotropic hormone; ACTH)-independent forms, in which aberrant adrenal receptors, through the binding of their respective ligands, could lead to chronic cortisol overproduction. They include receptors for gastric inhibitory peptide (GIP), beta-adrenergic agonists, luteinizing hormone (LH)/human chorionic gonadotropin, serotonin (5-HT(4) receptor), vasopressin (V(1) receptor), and angiotensin II (AT(1) receptor). In GIP-dependent Cushing syndrome, the most frequent subtype of ACTH-independent macronodular adrenal hyperplasia associated with the presence of aberrant adrenocortical hormone receptors described so far, octreotide administration before each meal showed clinical efficacy only in the first few months, probably because of somatostatin receptor downregulation in GIP secreting cells. Long-term medical treatments with propranolol and the gonadotropin-releasing hormone analog leuprorelin (leuprolide acetate) were effective in patients with catecholamine-dependent and LH-dependent Cushing syndrome, respectively. The oral vasopressin V(1) receptor antagonist OPC-21268 and the angiotensin II (AT(1)) receptor antagonist candesartan cilexetil were also able to decrease cortisol levels during the few days of administration of the drugs in patients with specific receptor abnormalities. These adrenal forms of Cushing syndrome are rare, and clinical data are scarce. Moreover, the real clinical significance of aberrant hormone receptors is still under investigation, as is the possibility of avoiding surgery by pharmacologic manipulation. Patients in whom these intriguing syndromes are suspected require detailed investigation protocols, which should be carried out in specialized centers. While awaiting further developments, the use of traditional medical treatment at the adrenal level with adrenal steroid inhibitors is still valuable in several instances. PMID- 15783248 TI - Clinical pharmacology of potent new bisphosphonates for postmenopausal osteoporosis. AB - Bisphosphonates are potent inhibitors of bone resorption, used in most bone diseases associated with high bone resorption levels. Several bisphosphonates, developed to prevent and treat postmenopausal osteoporosis, increase bone mineral density and decrease biochemical markers of bone turnover, and more importantly, reduce fracture risk. Alendronate and risedronate have proven their efficacy to reduce vertebral and hip fracture risk among postmenopausal osteoporotic women, using daily regimens. Weekly intermittent schedules, however, are now most commonly prescribed, because they have shown pharmacologic equivalence to the daily regimen. Ibandronate has been the first bisphosphonate to demonstrate vertebral fracture risk reduction using an intermittent regimen. Studies using ibandronate as intravenous injections every 3 months are under way. Zoledronic acid may also be an attractive option for the treatment of postmenopausal osteoporosis if a large ongoing trial proves that a single annual injection of this compound allows osteoporotic fracture risk reduction. PMID- 15783247 TI - The use of dehydroepiandrosterone therapy in clinical practice. AB - Dehydroepiandrosterone (DHEA) therapy is controversial due to sensationalized reports of epidemiologic studies and the over-the-counter availability of DHEA. Human clinical trials have investigated the potential efficacy of DHEA therapy in multiple conditions with resultant inconsistencies in findings. DHEA is unique compared with other adrenal steroids because of the fluctuation in serum levels found from birth into advancing age. The lower endogenous levels of DHEA and DHEA sulfate found in advancing age have been correlated with a myriad of health conditions. Also, some studies suggest gender-specific actions of endogenous and exogenous DHEA. We reviewed only pharmacokinetic studies and human clinical trials investigating the efficacy of DHEA therapy that were placebo-controlled as these provided the most reliable scientific basis for the evaluation of DHEA therapy. Pharmacodynamic studies suggest that doses of 30-50mg of oral DHEA may produce physiologic androgen levels, especially in women. These studies report a dose-dependent effect and lack of accumulation of serum androgen levels. Pharmacologic studies also reveal a gender-specific response to DHEA therapy such that testosterone levels are increased in women but not in men. Clinical trials suggest that 50mg of oral DHEA, but not <30mg, can increase serum androgen levels to within the physiologic range for young adults with primary and secondary adrenal insufficiency, possibly improve sexual function, improve mood and self esteem, and decrease fatigue/exhaustion. Whereas DHEA replacement therapy may be effective in treating patients with adrenal insufficiency, human clinical trials investigating its efficacy in conditions such as systemic lupus erythematosus, HIV, Alzheimer disease, advancing age, male sexual dysfunction, perimenopausal symptoms, depression, and cardiovascular disease have not provided consistent findings. PMID- 15783249 TI - Spotlight on orlistat in the management of patients with obesity. AB - Orlistat is an inhibitor of gastrointestinal lipases and, therefore, prevents the absorption of dietary fat. This agent reduced weight in obese adults and adolescents with or without co-morbidities (including type 2 diabetes mellitus, hypercholesterolemia, hypertension, metabolic syndrome) who received up to 4 years of therapy in conjunction with a hypocaloric diet. In obese patients, orlistat in combination with a hypocaloric diet improved metabolic risk factors and reduced the risk of developing type 2 diabetes. Furthermore, this agent was cost effective in patients with obesity, particularly those with type 2 diabetes. Orlistat is generally well tolerated, with gastrointestinal adverse events being most commonly reported. Orlistat, in addition to lifestyle and dietary intervention, is thus an attractive option for the treatment of patients with obesity, especially those with associated co-morbidities or at risk of developing type 2 diabetes. PMID- 15783250 TI - Creating locally relevant health information. PMID- 15783251 TI - Alas, poor Yorick: digging up the dead to make medical diagnoses. PMID- 15783253 TI - The coming of age of multicultural medicine. PMID- 15783252 TI - Protecting children from environmental toxins. PMID- 15783254 TI - Recurrent pleural and pericardial effusions due to sarcoidosis. PMID- 15783255 TI - Antifibrinolytic agents in traumatic haemorrhage. PMID- 15783256 TI - Hookworm: "the great infection of mankind". PMID- 15783257 TI - Exercise and health: can biotechnology confer similar benefits? PMID- 15783259 TI - Is it always unethical to use a placebo in a clinical trial? AB - BACKGROUND TO THE DEBATE: Placebos are used in trials to conceal whether a treatment is being given or not and hence to control for the psychosomatic effects of offering treatment. Placebo-controlled trials are controversial. Critics of such trials argue that if a proven effective therapy exists, a placebo should not be used. But proponents argue that placebo trials are still crucial to prove the efficacy and safety of many treatments. PMID- 15783258 TI - Noninvasive visualization of the activated alphavbeta3 integrin in cancer patients by positron emission tomography and [18F]Galacto-RGD. AB - BACKGROUND: The integrin alphavbeta3 plays an important role in angiogenesis and tumor cell metastasis, and is currently being evaluated as a target for new therapeutic approaches. Several techniques are being studied to enable noninvasive determination of alphavbeta3 expression. We developed [(18)F]Galacto RGD, a (18)F-labeled glycosylated alphavbeta3 antagonist, allowing monitoring of alphavbeta3 expression with positron emission tomography (PET). METHODS AND FINDINGS: Here we show by quantitative analysis of images resulting from a small animal PET scanner that uptake of [(18)F]Galacto-RGD in the tumor correlates with alphavbeta3 expression subsequently determined by Western blot analyses. Moreover, using the A431 human squamous cell carcinoma model we demonstrate that this approach is sensitive enough to visualize alphavbeta3 expression resulting exclusively from the tumor vasculature. Most important, this study shows, that [(18)F]Galacto-RGD with PET enables noninvasive quantitative assessment of the alphavbeta3 expression pattern on tumor and endothelial cells in patients with malignant tumors. CONCLUSIONS: Molecular imaging with [(18)F]Galacto-RGD and PET can provide important information for planning and monitoring anti-angiogenic therapies targeting the alphavbeta3 integrins and can reveal the involvement and role of this integrin in metastatic and angiogenic processes in various diseases. PMID- 15783260 TI - Serum uric acid and coronary heart disease in 9,458 incident cases and 155,084 controls: prospective study and meta-analysis. AB - BACKGROUND: It has been suggested throughout the past fifty years that serum uric acid concentrations can help predict the future risk of coronary heart disease (CHD), but the epidemiological evidence is uncertain. METHODS AND FINDINGS: We report a "nested" case-control comparison within a prospective study in Reykjavik, Iceland, using baseline values of serum uric acid in 2,456 incident CHD cases and in 3,962 age- and sex-matched controls, plus paired serum uric acid measurements taken at baseline and, on average, 12 y later in 379 participants. In addition, we conducted a meta-analysis of 15 other prospective studies in eight countries conducted in essentially general populations. Compared with individuals in the bottom third of baseline measurements of serum uric acid in the Reykjavik study, those in the top third had an age- and sex-adjusted odds ratio for CHD of 1.39 (95% confidence interval [CI], 1.23-1.58) which fell to 1.12 (CI, 0.97-1.30) after adjustment for smoking and other established risk factors. Overall, in a combined analysis of 9,458 cases and 155,084 controls in all 16 relevant prospective studies, the odds ratio was 1.13 (CI, 1.07-1.20), but it was only 1.02 (CI, 0.91-1.14) in the eight studies with more complete adjustment for possible confounders. CONCLUSIONS: Measurement of serum uric acid levels is unlikely to enhance usefully the prediction of CHD, and this factor is unlikely to be a major determinant of the disease in general populations. PMID- 15783261 TI - Tumor cell recognition efficiency by T cells. PMID- 15783262 TI - T cell epitope immunotherapy induces a CD4+ T cell population with regulatory activity. AB - BACKGROUND: Synthetic peptides, representing CD4(+) T cell epitopes, derived from the primary sequence of allergen molecules have been used to down-regulate allergic inflammation in sensitised individuals. Treatment of allergic diseases with peptides may offer substantial advantages over treatment with native allergen molecules because of the reduced potential for cross-linking IgE bound to the surface of mast cells and basophils. METHODS AND FINDINGS: In this study we address the mechanism of action of peptide immunotherapy (PIT) in cat allergic, asthmatic patients. Cell-division-tracking dyes, cell-mixing experiments, surface phenotyping, and cytokine measurements were used to investigate immunomodulation in peripheral blood mononuclear cells (PBMCs) after therapy. Proliferative responses of PBMCs to allergen extract were significantly reduced after PIT. This was associated with modified cytokine profiles generally characterised by an increase in interleukin-10 and a decrease in interleukin-5 production. CD4(+) cells isolated after PIT were able to actively suppress allergen-specific proliferative responses of pretreatment CD4(neg) PBMCs in co culture experiments. PIT was associated with a significant increase in surface expression of CD5 on both CD4(+) and CD8(+) PBMCs. CONCLUSION: This study provides evidence for the induction of a population of CD4(+) T cells with suppressor/regulatory activity following PIT. Furthermore, up-regulation of cell surface levels of CD5 may contribute to reduced reactivity to allergen. PMID- 15783263 TI - Cholesterol, statins, and Alzheimer disease. PMID- 15783264 TI - Accounting for individual differences in risk of Alzheimer disease. PMID- 15783265 TI - A Canadian perspective. PMID- 15783266 TI - Statement of principles for health care journalists. PMID- 15783268 TI - How does PLoS medicine manage competing interests? PMID- 15783272 TI - Stage of change transitions and processes of change, decisional balance, and self efficacy in smokers: a transtheoretical model validation using longitudinal data. AB - Interactions were examined between stage of change transitions and intraindividual increases or decreases in the processes of change, pros and cons of smoking, and situational temptations longitudinally. A total of 786 ever smokers was assessed 2 times, 6 months apart, with respect to the transtheoretical model (TTM) constructs. Two significant discriminant functions within initial precontemplators and 1 significant function within initial contemplators were found. Ten out of 15 TTM variables contributed to at least 1 function. The functions mainly distinguished between preabstinence (precontemplation, contemplation, or preparation) and abstinence (action or maintenance) stages of change, that is, between current and former smokers. This is one of the few studies providing a longitudinal validation of the postulates of the TTM. PMID- 15783273 TI - Predicting alcohol patterns in first-year college students through motivational systems and reasons for drinking. AB - In the present study, a typological approach was used to identify patterns of alcohol use in a sample of 533 college freshmen students (<21 years old; 342 women; 191 men), on the basis of quantity and frequency of consumption, and alcohol-related problems. Personality (sensitivity to reward, SR; sensitivity to punishment, SP) and reasons for drinking were examined as correlates of drinking patterns through a mediational model. Analyses were done separately by gender. Latent profile analyses suggested 5 drinking patterns for both genders, including 3 problematic groups. SR was only associated with the problematic drinking patterns, and enhancement, coping, and social reasons for drinking mediated this relationship. These findings demonstrate the utility of latent profile analysis for identifying a drinking typology and for integrating personality and drinking motives to distinguish drinking patterns. PMID- 15783274 TI - Elementary school drinking: the role of temperament and learning. AB - Drinking in elementary school, despite its low base rate, has been shown to predict alcohol use in middle school (Wilson, Battistich, Syme, & Boyce, 2002), which in turn predicts alcohol abuse or dependence in young adults (Guo, Collins, Hill, & Hawkins, 2000). The authors report 1 of the 1st examinations of the relationship between personality and psychosocial learning risk factors and drinking behavior among elementary school students. Fifth-grade students completed measures of disinhibition, positive and negative alcohol expectancies, and drinking. MIMIC modeling, tests of mediation, and tests of moderation were completed to test these relations. It was found that disinhibition and positive alcohol expectancies were each related to drinking in 5th graders. Disinhibition moderated the relation between positive alcohol expectancies and drinking in some cases. Mediation was not supported. PMID- 15783275 TI - Relapse in alcohol- and drug-disordered adolescents with comorbid psychopathology: changes in psychiatric symptoms. AB - This study examined reciprocal relationships between posttreatment substance use and psychiatric symptoms in adolescents with both a substance use disorder and an Axis I mental health disorder. Participants (13-18 years old) were recruited from inpatient treatment centers and interviewed during treatment and monthly for 6 months. Participants who relapsed (N=103; 48% female) reported the incidence and severity of psychiatric symptoms experienced before and after their 1st posttreatment substance use. The number of symptoms and depression symptoms experienced were related to use of stimulants and other drugs during relapse. There was evidence for both self-medication (symptom reduction) and rebound (symptom exacerbation) effects of substance use on symptom severity. These results demonstrate that, for adolescents with both substance use and mental health disorders, psychiatric symptoms are 1 factor influencing posttreatment substance use. PMID- 15783276 TI - Alcohol consumption and intimate partner violence by alcoholic men: comparing violent and nonviolent conflicts. AB - Alcoholic men and their relationship partners were interviewed about a conflict in which physical assault occurred and 1 in which psychological aggression occurred without physical assault. The interview assessed the quantity of alcohol consumed prior to each conflict, other drug use, and the topics, location, timing, duration, and speed of escalation for each conflict. The number of standard drinks consumed by the husband in the previous 12 hr was significantly higher prior to violent versus nonviolent conflicts for both self- and collateral reports, as was blood alcohol concentration estimated from self-report. Other drug use was not significantly different. Greater drinking by wives prior to violent conflicts was found in some analyses. These within-subject comparisons help to rule out individual difference explanations for the alcohol-violence association and indicate that alcohol consumption is a proximal risk factor for partner violence in alcoholic men. PMID- 15783277 TI - Associations among childhood trauma, adolescent problem behaviors, and adverse adult outcomes in substance-abusing women offenders. AB - This article explores relationships among exposure to childhood abuse and traumatic events, adolescent conduct problems and substance abuse, and adult psychological distress and criminal behaviors in a sample of substance-abusing women offenders (N=440). Latent variable structural equation models revealed direct relationships between several childhood traumatic events and greater adolescent conduct problems and substance abuse. Conduct problems predicted more adult criminal behavior, and adolescent substance abuse predicted higher levels of current psychological distress. There were direct relationships between several types of traumatic events and current psychological distress and between traumatic events and specific criminal behaviors. Ethnic differences were also found, suggesting different pathways to criminal behavior. The findings underscore the need to provide trauma-related services for substance-abusing women offenders. PMID- 15783278 TI - Conversion and validation of the Teen-Addiction Severity Index (T-ASI) for Internet and automated-telephone self-report administration. AB - This study converted the Teen-Addiction Severity Index (T-ASI) into self-report formats using Internet (Net) and interactive voice response (IVR) automated telephone technologies. Reliability and convergent validity were assessed among 95 inpatient adolescent participants. Current functioning scores obtained by clinician interview correlated well with self-report Net (mean r=.74, SD=.14) and IVR (mean r=.72, SD=.16). Lifetime history items obtained by clinicians were consistent with self-report Net (mean r=.60, SD=.32; mean kappa=.67, SD=.24) and IVR formats (mean r=.60, SD=.30; mean kappa=.64, SD=.26). Participants rated "ease of use" as being high for both Net and IVR formats. These findings suggest that automated T-ASI administration is a valid and potentially less expensive alternative to clinician-administered T-ASI interviews. PMID- 15783279 TI - The stability and impact of environmental factors on substance use and problems after adolescent outpatient treatment for cannabis abuse or dependence. AB - Because alcohol or other drug use following adolescent substance abuse treatment is common, understanding mediators of posttreatment outcome could help improve treatment interventions. The authors conducted path analyses based on data from 552 adolescents (aged 12-18; 82% male) with cannabis abuse or dependence who participated in outpatient treatment. The analysis used the Family Conflict and Cohesion subscales, from the Family Environment Scale, and several scales and indices from the Global Appraisal of Individual Needs. Family conflict, family cohesion, and social support indirectly predicted substance use and substance related problems as mediated by recovery environment and social risk. This model replicated across 4 follow-up waves (3, 6, 9, and 12 months postintake). These results support the idea of targeting environmental factors during continuing care as a way to improve treatment outcomes for adolescents with cannabis disorders. PMID- 15783280 TI - Indicators of client engagement: influences on alcohol treatment satisfaction and outcomes. AB - This investigation explored the relationship of client engagement variables (client expectations, therapeutic/working alliance, and session attendance) with treatment satisfaction and posttreatment drinking-related outcomes using data from 2 outpatient alcohol treatment studies (N=208). Path analysis was used to test a model in which engagement variables jointly influence client satisfaction with treatment and subsequent drinking-related outcomes. The proposed model fit well with the data and accounted for 14-23% of the variance in posttreatment outcomes. The relationships in the model suggest that the link between treatment satisfaction and outcome is clarified by examining client engagement variables, which relate indirectly to outcome by means of client satisfaction. PMID- 15783281 TI - Motivational interventions for heavy drinking college students: examining the role of discrepancy-related psychological processes. AB - The authors examined the effects of a brief motivational intervention for heavy, episodic alcohol use on discrepancy-related psychological processes. Heavy drinking college students (N=73) were randomly assigned to a motivationally based intervention (MBI) or an assessment-only control (AC) condition. Cognitive (actual-ideal discrepancy) and affective (2 forms of cognitive dissonance) discrepancy processes were assessed at baseline and immediately following the experimental manipulation. At 6-week follow-up, MBI participants demonstrated significantly greater reductions in problematic drinking than AC participants. Moreover, actual-ideal discrepancy and negative, self-focused dissonance were significantly increased following the intervention (discomfort-related dissonance was not) and were correlated with outcome alcohol involvement. These discrepancy processes did not, however, mediate the relationship between condition and outcome. The findings lend some support to the role of discrepancy enhancement in drinking-related behavior change among college students. PMID- 15783282 TI - Effects of smoking urge on temporal cognition. AB - The authors examined temporal aspects of smoking urge. In Experiment 1, smokers assigned to high- or low-urge conditions were informed they would be allowed to smoke in 2.5 min. They next completed measures of time perception. High-urge smokers reported 45 s to pass significantly more slowly than did low-urge smokers. In Experiment 2, the high-urge smokers from Experiment 1 anticipated that their urges would climb steadily over the next 45 min if they were not permitted to smoke. Another group of high-urge smokers actually reported their urges over 45 min. These urge ratings did not show the steady rise anticipated by the first group. Results suggest that smoking urge may affect time perception and that craving smokers overpredict the duration and intensity of their own future smoking urges if they abstain. PMID- 15783283 TI - Heart rate increase to alcohol administration and video lottery terminal (VLT) play among regular VLT players. AB - The authors examined heart rate responses to video lottery terminal (VLT) play and alcohol intake. Forty-four VLT players were randomized to an alcohol (mean blood alcohol concentration=0.06%) or a control beverage condition. Heart rate was recorded at pre- and postdrinking baseline and during VLT play. Alcohol participants displayed elevated heart rates relative to controls at postdrinking and VLT play. Controls displayed elevated heart rates during VLT play relative to both pre- and postdrinking baselines, whereas alcohol participants displayed elevations at post- relative to predrinking and at VLT play relative to postdrinking. Heart rate increases from predrinking to VLT play were greater among alcohol participants relative to controls. Results provide novel information that the combination of VLT play and alcohol further intensifies heart rate increase relative to either alone. Implications for pathological gambling and alcohol use disorder comorbidity are discussed. PMID- 15783284 TI - Alcohol use in college students: effects of level of self-esteem, narcissism, and contingencies of self-worth. AB - The unique effects of level of self-esteem, narcissism, and contingencies of self worth assessed prior to college on alcohol use during the freshman year were examined in a longitudinal study of 620 college students. Narcissism predicted alcohol use, but level of self-esteem did not. Basing self-worth on appearance predicted more alcohol use, whereas the virtue, God's love, and academic competence contingencies predicted less alcohol use, independent of other personality measures and joining a sorority or a fraternity. Further, the virtue and academic competence contingencies were associated with decreases in alcohol use from the 1st to the 2nd semester. PMID- 15783285 TI - Pathways of youth gambling problem severity. AB - Prospective studies are needed to advance knowledge of the developmental features of gambling involvement and associated problems. Developmental pathways of youth gambling problem severity (no problem gambling, at-risk gambling, and problem gambling) are described on the basis of a 3-wave data set that spans midadolescence to young adulthood (N=305). The most prevalent group was the resistors (no problem gambling at all data points); 60% of study participants were in this group. New incidence cases (no problem gambling followed by at-risk or problem gambling) and desistors (at-risk or problem gambling followed by no problem gambling) were found among 21% and 13% of participants, respectively. Only 4% of cases were persistors, that is, at-risk or problem gambling at all 3 data waves. Findings are discussed in light of extant research on adolescent gambling that heretofore has not benefited from a developmental pathway perspective. PMID- 15783286 TI - Alcohol's effect on triggered displaced aggression. AB - The authors examined alcohol's effect on triggered displaced aggression, the hostile reaction to a second provoking person after provocation from a first. Participants consumed an alcoholic or a nonalcoholic beverage. Subsequently, one individual provoked all of them with moderate intensity. Then, 2 groups were studied: those who received or who failed to receive a second provocation of minimal intensity. Consistent with prior research, participants who received a second, minimal provocation displayed more aggression than those who did not. After participants drank alcohol, the magnitude of this difference was significantly greater, indicating that alcohol increases triggered displaced aggression. PMID- 15783287 TI - Performance of alcohol and safer sex change rulers compared with readiness to change questionnaires. AB - As part of a larger intervention study, the authors hypothesized that change rulers created for alcohol and safer sex would be equivalent to longer questionnaires. Ninety-six male college students completed rulers and questionnaires for assessing behavior change readiness. Participants' scores on the rulers significantly correlated with their scores on the questionnaires (r=.77 for alcohol; r=.77 for safer sex). In both domains, the rulers outperformed the questionnaires in predicting behavioral intentions, suggesting that the rulers had at least comparable concurrent criterion validity. This finding is the first of its kind in the safe sex literature and suggests that quick assessments of readiness to change are possible. Because the rulers are a continuous measure, the results are consistent with the idea that the change process is continuous rather than a series of discrete stages. PMID- 15783288 TI - A neural theory of visual attention: bridging cognition and neurophysiology. AB - A neural theory of visual attention (NTVA) is presented. NTVA is a neural interpretation of C. Bundesen's (1990) theory of visual attention (TVA). In NTVA, visual processing capacity is distributed across stimuli by dynamic remapping of receptive fields of cortical cells such that more processing resources (cells) are devoted to behaviorally important objects than to less important ones. By use of the same basic equations used in TVA, NTVA accounts for a wide range of known attentional effects in human performance (reaction times and error rates) and a wide range of effects observed in firing rates of single cells in the primate visual system. NTVA provides a mathematical framework to unify the 2 fields of research--formulas bridging cognition and neurophysiology. PMID- 15783289 TI - Approximate optimal control as a model for motor learning. AB - Current models of psychological development rely heavily on connectionist models that use supervised learning. These models adapt network weights when the network output does not match the target outputs computed by some agent. The authors present a model of motor learning in which the child uses exploration to discover appropriate ways of responding. The model is consistent with what is known about how neural systems evaluate behavior. The authors model the development of reaching and investigate N. Bernstein's (1967) hypotheses about early motor learning. Simulations show the course of learning as well as model the kinematics of reaching by a dynamical arm. PMID- 15783290 TI - From the lexicon to expectations about kinds: a role for associative learning. AB - In the novel noun generalization task, 2 1/2-year-old children display generalized expectations about how solid and nonsolid things are named, extending names for never-before-encountered solids by shape and for never-before encountered nonsolids by material. This distinction between solids and nonsolids has been interpreted in terms of an ontological distinction between objects and substances. Nine simulations and behavioral experiments tested the hypothesis that these expectations arise from the correlations characterizing early learned noun categories. In the simulation studies, connectionist networks were trained on noun vocabularies modeled after those of children. These networks formed generalized expectations about solids and nonsolids that match children's performances in the novel noun generalization task in the very different languages of English and Japanese. The simulations also generate new predictions supported by new experiments with children. Implications are discussed in terms of children's development of distinctions between kinds of categories and in terms of the nature of this knowledge. PMID- 15783291 TI - The dynamics of scaling: a memory-based anchor model of category rating and absolute identification. AB - A memory-based scaling model--ANCHOR--is proposed and tested. The perceived magnitude of the target stimulus is compared with a set of anchors in memory. Anchor selection is probabilistic and sensitive to similarity, base-level strength, and recency. The winning anchor provides a reference point near the target and thereby converts the global scaling problem into a local comparison. An explicit correction strategy determines the final response. Two incremental learning mechanisms update the locations and base-level activations of the anchors. This gives rise to sequential, context, transfer, practice, and other dynamic effects. The scale unfolds as an adaptive map. A hierarchy of models is tested on a battery of quantitative measures from 2 experiments in absolute identification and category rating. PMID- 15783292 TI - Life stress, the "kindling" hypothesis, and the recurrence of depression: considerations from a life stress perspective. AB - Major depression is frequently characterized by recurrent episodes over the life course. First lifetime episodes of depression, however, are typically more strongly associated with major life stress than are successive recurrences. A key theoretical issue involves how the role of major life stress changes from an initial episode over subsequent recurrences. The primary conceptual framework for research on life stress and recurrence of depression is the "kindling" hypothesis (R. M. Post, 1992). Despite the strengths of the kindling hypothesis, a review of the research literature reveals inconsistencies and confusion about life stress and its implications for the recurrence of depression. Adopting a life stress perspective, the authors introduce 3 major themes that resolve the inconsistencies in the current literature. They integrate these themes and extrapolate the ideas with available data to develop a preliminary framework for evaluating competing explanatory models and to guide research on life stress and the recurrence of depression. PMID- 15783293 TI - Imaginary relish and exquisite torture: the elaborated intrusion theory of desire. AB - The authors argue that human desire involves conscious cognition that has strong affective connotation and is potentially involved in the determination of appetitive behavior rather than being epiphenomenal to it. Intrusive thoughts about appetitive targets are triggered automatically by external or physiological cues and by cognitive associates. When intrusions elicit significant pleasure or relief, cognitive elaboration usually ensues. Elaboration competes with concurrent cognitive tasks through retrieval of target-related information and its retention in working memory. Sensory images are especially important products of intrusion and elaboration because they simulate the sensory and emotional qualities of target acquisition. Desire images are momentarily rewarding but amplify awareness of somatic and emotional deficits. Effects of desires on behavior are moderated by competing incentives, target availability, and skills. The theory provides a coherent account of existing data and suggests new directions for research and treatment. PMID- 15783294 TI - Reasoning about relations. AB - Inferences about spatial, temporal, and other relations are ubiquitous. This article presents a novel model-based theory of such reasoning. The theory depends on 5 principles. (a) The structure of mental models is iconic as far as possible. (b) The logical consequences of relations emerge from models constructed from the meanings of the relations and from knowledge. (c) Individuals tend to construct only a single, typical model. (d) They spontaneously develop their own strategies for relational reasoning. (e) Regardless of strategy, the difficulty of an inference depends on the process of integration of the information from separate premises, the number of entities that have to be integrated to form a model, and the depth of the relation. The article describes computer implementations of the theory and presents experimental results corroborating its main principle. PMID- 15783295 TI - The robust beauty of majority rules in group decisions. AB - How should groups make decisions? The authors provide an original evaluation of 9 group decision rules based on their adaptive success in a simulated test bed environment. When the adaptive success standard is applied, the majority and plurality rules fare quite well, performing at levels comparable to much more resource-demanding rules such as an individual judgment averaging rule. The plurality rule matches the computationally demanding Condorcet majority winner that is standard in evaluations of preferential choice. The authors also test the results from their theoretical analysis in a behavioral study of nominal human group decisions, and the essential findings are confirmed empirically. The conclusions of the present analysis support the popularity of majority and plurality rules in truth-seeking group decisions. PMID- 15783296 TI - Accounting for belief bias in a mental model framework: comment on Klauer, Musch, and Naumer (2000). AB - K. C. Klauer, J. Musch, and B. Naumer (2000; see record 2000-02818-008) presented a general multinomial model of belief bias effects in syllogistic reasoning. They claimed to map a particular mental model account of belief bias (J. V. Oakhill, P. N. Johnson-Laird, & A. Garnham, 1989; see record 1989-38845-001)) onto this model and to show empirically that it is incorrect. The authors argue that this mental model account does not map onto the multinomial model and that it can account for the data presented by Klauer et al. (Experiments 1-4). The authors further argue that additional data Klauer et al. presented in support of a new model of their own (Experiments 5-8) are explained by this mental model account. The mental model account is, therefore, refuted neither by Klauer et al.'s theoretical analysis nor by any of the results they presented. Furthermore, the account can accommodate more recent findings on belief bias in a more satisfactory way than can alternative models that have been proposed. PMID- 15783298 TI - An unusual anti-H lectin inhibited by milk from individuals with the Bombay phenotype. AB - There are several lectins with anti-H specificity but few of them serve as useful reagents. An anti-H lectin, extracted from the seeds of the plant Momordica dioica Roxb. ex willd., was tested for its hemagglutination and inhibition properties, using standard serologic methods and panel RBCs, serum, saliva, milk, and oligosaccharides purified from milk. The extract displayed strongest agglutination with group O RBCs and was weakest with group A1B RBCs in a spectrum of O>A2>B>A2B>A1>A1B; the extract failed to react with the RBCs from 25 individuals with the Bombay (Oh) phenotype and was inhibited by H secretor saliva, hence it was characterized as anti-H. However, its inhibition by milk samples from five mothers with the Bombay phenotype called into question its specificity as anti-H. The lectin reacted as strongly with group O ii (adult) RBCs as with normal OI RBCs, ruling out its specificity as anti-HI. Hemagglutination inhibition was observed with 2'-fucosyllactose (Type 2 H) and lacto-N-fucopentose-I (Type 1 H), suggesting that the binding of the lectin had preference for H structures. However, inhibition by N-acetyllactosamine, lacto Ntetraose, and lacto-N-neotetraose suggested that the lectin also recognized unsubstituted terminal beta-linked galactose units. The hemagglutinin property in the present lectin showed an unusual anti-H specificity. The lectin was inhibited by milk from Bombay phenotype individuals and certain milk oligosaccharides not specific for the H antigen. PMID- 15783299 TI - Gene frequencies of the HPA-1 to 6 and Gov human platelet antigens in Thai blood donors. AB - Human platelet alloantigens (HPA) are important in neonatal alloimmune thrombocytopenia (NAIT), posttransfusion purpura (PTP), platelet transfusion refractoriness, passive alloimmune thrombocytopenia, and transplantation associated alloimmune thrombocytopenia. Thus, HPA genotyping is essential in diagnosis and treatment. We analyzed HPA-1 to 6 and Gov alleles, using PCR with sequence specific primers (PCR-SSP) in 500 Thai blood donors who had been HLA class I antigen typed. HPA-4a was present in all samples. HPA-1b, -2b, -5b, and 6b were rare, and HPA-4b was not found. HPA-3a and -3b showed frequencies of 56.0 percent and 44.0 percent, respectively. Gova and Govb showed frequencies of 49.1 percent and 50.9 percent, respectively. The prevalence rates of HPA-1 to 6 gene frequencies (GFs) were consistent with those of other Asian populations rather than those of Caucasians. We also report on the GFs of Gova and Govb, which also are comparable to those of Asian populations. Our results could establish a useful HPA- and HLA-matched plateletpheresis donor file and provide an improvement of platelet alloantibody detection in alloimmune thrombocytopenic patients, and, therefore, a more effective platelet transfusion program. PMID- 15783300 TI - ABO, Rh, MNS, Duffy, Kidd,Yt, Scianna, and Colton blood group systems in indigenous Chinese. AB - The frequencies of selected alleles in the ABO, Rh, MNS, Duffy, Kidd, Yt, Scianna, and Colton blood group systems were determined among four indigenous Chinese ethnic populations: Han, Tajik, She, and Yugu. Genotypes were determined by PCR or PCR with sequence specific primers (PCR-SSP). In the Han population, the frequencies of A1, A2, B, and O1 alleles were 0.189, 0.003, 0.170, and 0.638, respectively, and the O2 allele was not identified. Among D+ Hans, the frequencies of C and c alleles were 0.67 and 0.33 and the frequencies of E and e were 0.22 and 0.78, respectively. Among D- Hans, the frequencies of C and c alleles were 0.23 and 0.77 and the frequencies of E and e were 0.04 and 0.96, respectively. The frequencies of M and N alleles were 0.478 and 0.522 among Hans and 0.655 and 0.345 among Tajiks, respectively. The frequencies of Fya and Fyb alleles were 0.94 and 0.06 among Hans and 0.98 and 0.02 among Shes, respectively. The frequencies of Jka and Jkb alleles were 0.49 and 0.51 among Hans and 0.56 and 0.44 among Shes, respectively. The frequency of the Yta allele was 1.00 among Hans. The frequencies of Yta and Ytb alleles were 0.94 and 0.06 among Tajiks, respectively. The frequency of the Sc1 allele was 1.00 in both Han and Tajik ethnic populations. The frequency of the Coa allele was 1.00 in Han, She, and Tajik ethnic populations. PMID- 15783301 TI - Expression of Duffy antigen receptor for chemokines during reticulocyte maturation: using a CD71 flow cytometric technique to identify reticulocytes. AB - Flow cytometric methods commonly used to identify reticulocytes are of limited usefulness in malarious areas, since RNA staining also detects plasmodia. An important antigen expressed on reticulocytes is Duffy antigen receptor for chemokines (DARC, also known as Fy), the receptor for Plasmodium vivax. An early marker for reticulocytes is CD71 (transferrin receptor). We have been interested in CD71 as an alternative marker for reticulocytes in the context of Fy expression. Flow cytometry was used to determine the expression of Fy on CD71 positive and -negative reticulocytes and to correlate serology and genotype. A reduction of 13 percent was seen in Fy6 expression between CD71-positive reticulocytes and RNA-positive reticulocytes. CD71 disappears early during reticulocyte maturation, while Fy6 expression is relatively preserved. CD71 is an alternative to staining for RNA for reticulocyte assays relating to Fy6 expression. PMID- 15783302 TI - Rh antigens and phenotype frequencies of the Ibibio, Efik, and Ibo ethnic nationalities in Calabar, Nigeria. AB - This report forms part of the study on the Rh phenotypes within the various ethnic nationalities in the south-south region of Nigeria. The aim is to demonstrate the Rh polymorphisms among the people of African descent. The frequencies of Rh blood group antigens and phenotypes of the Ibibio, Efik, and Ibo ethnic nationalities in Calabar municipality, Nigeria, were determined using standard serologic techniques. Of the 720 Calabar individuals tested, the frequencies of the Rh antigens within the nationalities were c (100%), e (96.38%), D (96.38%), E (15.22%), and C (3.62%) for the Ibibios; c (100%), e (95.60%), D (96.70%), E (21.98%), and C (0%) for the Efiks; and c (100%), e (94.29%), D (91.43%), E (28.57%), and C (2.86%) for the Ibos. The overall frequencies of the Rh antigens in these 720 individuals were c (100%), e (95.56%), D (94.44%), E (18.89%), and C (2.78%). Forty (5.56%) were found to be D , while all were found to possess the c antigen. The most frequently occurring Rh phenotype was Dccee, with a frequency of 73.61 percent. The alternative allele, C, did not appear in homozygous form (CC) in the population tested. This study further demonstrates the variability of Rh blood group phenotypes in Nigeria and Africa. PMID- 15783303 TI - FCGR3B polymorphism in three ethnic Chinese populations. AB - Fc gamma RIIIb receptor is expressed primarily on neutrophils as three polymorphic antigens (HNA-1a, HNA-1b, and HNA-1c) that are encoded by alleles FCGR3B*1, FCGR3B*2, and FCGR3B*3, respectively. These antigens play an important role in immune neutropenia; their absence predisposes individuals who lack them to life-threatening infections. This study investigated the FCGR3B gene frequencies in three ethnic Chinese populations: Han, She, and Tajik. FCGR3B*1, FCGR3B*2, and FCGR3B*3 were genotyped by PCR using sequence specific primers (PCR SSP). The results showed the gene frequencies were 0.55 for FCGR3B*1 and 0.45 for FCGR3B*2 in 177 Han individuals, 0.69 for FCGR3B*1 and 0.31 for FCGR3B*2 in 87 She individuals, and 0.35 for FCGR3B*1 and 0.65 for FCGR3B*2 in 99 Tajik individuals, respectively. The FCGR3Bnull genotype was not found, but the FCGR3B*3 allele was identified in only three individuals in the Tajik population. DNA clone and sequencing confirmed that these individuals had the C ==>A mutation at position 266 on exon 3. This study found that the gene frequencies in Han and She ethnic groups were similar to those previously reported in the Asian population, but the FCGR3B allele frequencies in the Tajik population were more similar to that of Caucasians. PMID- 15783305 TI - Generalized fluctuation relation and effective temperatures in a driven fluid. AB - By numerical simulation of a Lennard-Jones-like liquid driven by a velocity gradient gamma we test the fluctuation relation (FR) below the (numerical) glass transition temperature T(g) . We show that, in this region, the FR deserves to be generalized introducing a numerical factor X (T, gamma) <1 that defines an "effective temperature" T(FR) =T/X . On the same system we also measure the effective temperature T(eff) , as defined from the generalized fluctuation dissipation relation, and find a qualitative agreement between the two different nonequilibrium temperatures. PMID- 15783306 TI - Decomposition of force fluctuations far from equilibrium. AB - By studying a nonequilibrium Langevin system, we find that a simple condition determines the decomposition of the coarse-grained force into a dissipative force, an effective driving force and noise. From this condition, we derive a universal inequality, D > or = gamma mu2(d)T , relating the diffusion constant D , the differential mobility mu(d) , the bare friction constant gamma and the temperature T . Due to the general nature of the argument we present, we believe that our idea concerning this decomposition can be applied to a wide class of systems far from equilibrium. PMID- 15783307 TI - Interplay of instabilities in mounded surface growth. AB - We numerically study a one-dimensional conserved growth equation with competing linear (Ehrlich-Schwoebel) and nonlinear instabilities. As a control parameter is varied, this model exhibits a nonequilibrium phase transition between two mounded states, one of which exhibits slope selection and the other does not. The coarsening behavior of the mounds in these two phases is studied in detail. In the absence of noise, the steady-state configuration depends crucially on which of the two instabilities dominates the early time behavior. PMID- 15783308 TI - Temperature variation of film tension above the bulk smectic-A-isotropic transition in freestanding liquid-crystal films. AB - We have measured the temperature variation of film tension above the bulk smectic A-isotropic transition in freestanding films of two liquid-crystal compounds. Above the transition, the tension increases sharply with temperature, and the slope is proportional to the film's thickness regardless of whether or not the compound exhibits regular layer-by-layer thinning. The data can be interpreted in the context of theoretical models for layer thinning. PMID- 15783309 TI - Real-time microbe detection based on director distortions around growing immune complexes in lyotropic chromonic liquid crystals. AB - We describe director distortions in the nematic liquid crystal (LC) caused by a spherical particle with tangential surface orientation of the director and show that light transmittance through the distorted region is a steep function of the particle's size. The effect allows us to propose a real-time microbial sensor based on a nontoxic lyotropic chromonic LC (LCLC) that detects and amplifies the presence of immune complexes. A cassette is filled with LCLC, antibody, and antigen-bearing particles. Small and isolated particles cause no macroscopic distortions of the LCLC. Upon antibody-antigen binding, the growing immune complexes distort the director and cause detectable optical transmittance between crossed polarizers. PMID- 15783310 TI - Dynamics of the antibody-T. cruzi competition during Chagas infection: prognostic relevance of intracellular replication. AB - A recently proposed model for the competitive parasite-antibody interactions in Chagas disease is extended by separately describing the parasitic intracellular and extracellular phases. The model solutions faithfully reproduce available population data and yield predictions for parasite-induced cardiac cell damage. PMID- 15783311 TI - Evidence of universality for the May-Wigner stability theorem for random networks with local dynamics. AB - We consider a random network of nonlinear maps exhibiting a wide range of local dynamics, with the links having normally distributed interaction strengths. The stability of such a system is examined in terms of the asymptotic fraction of nodes that persist in a nonzero state. Scaling results show that the probability of survival in the steady state agrees remarkably well with the May-Wigner stability criterion derived from linear stability arguments. This suggests universality of the complexity-stability relation for random networks with respect to arbitrary global dynamics of the system. PMID- 15783312 TI - Symmetric Brownian motor. AB - In this paper, we present a model of a symmetric Brownian motor which changes the sign of its velocity when the temperature gradient is inverted. The velocity, external work, and efficiency are studied as a function of the temperatures of the baths and other relevant parameters. The motor shows a current reversal when another parameter (a phase shift) is varied. Analytical predictions and results from numerical simulations are performed and agree very well. Generic properties of this type of motor are discussed. PMID- 15783313 TI - Enhancement of coherent response by quenched disorder. AB - We investigate the effects of quenched disorder on the coherent response in a driven system of coupled oscillators. In particular, the interplay between quenched noise and periodic driving is explored, with particular attention to the possibility of resonance. The phase velocity is examined as the response of the system; revealed is the enhancement of the fraction of oscillators locked to the periodic driving, displaying resonance behavior. It is thus concluded that resonance behavior may also be induced by quenched disorder that does not have time-dependent correlations. PMID- 15783314 TI - Window effect in a discretized model for diffusion of a chain in one dimension. AB - We introduce a model to study the diffusion of chains in microporous solids. The difficulties a chain has to escape from a pore where it is confined is found to strongly depend on the ratio between the chain length and the cage size. This dynamic effect implies a nonstandard behavior of the diffusion coefficient. We found a window effect that can be explained without using any energy argument. PMID- 15783315 TI - Inequivalence of statistical ensembles in single molecule measurements. AB - We study the role of fluctuations in single molecule experimental measurements of force-extension (f-zeta) curves. We use the worm-like chain (WLC) model to bring out the connection between the Helmholtz ensemble characterized by the free energy [F(zeta)] and the Gibbs ensemble characterized by the free energy [G(f)] . We consider the rigid rod limit of the WLC model as an instructive special case to bring out the issue of ensemble inequivalence. We point out the need for taking into account the free energy of transition when one goes from one ensemble to another. We also comment on the "phase transition" noticed in an isometric setup for semiflexible polymers and propose a realization of its thermodynamic limit. We present general arguments which rule out nonmonotonic force-extension curves in some ensembles and note that these do not apply to the isometric ensemble. PMID- 15783316 TI - Virial coefficients of hard spheres and hard disks up to the ninth. AB - A technique for topological analysis of the Ree-Hoover diagrams is developed with the aim to calculate the Ree-Hoover weights up to the ninth order with moderate demands on computer storage and CPU time. The ninth virial coefficients of hard spheres and disks are calculated, and the lower virial coefficients are accurately recalculated. The calculations require several spanning diagrams; the most important spanning chains are generated by reptation, other spanning diagrams by the standard Metropolis Monte Carlo algorithm. The tenth and eleventh virial coefficients for hard spheres are estimated. PMID- 15783317 TI - Critical viscosity exponent for classical fluids. AB - A self-consistent mode-coupling calculation of the critical viscosity exponent z(eta) for classical fluids is performed by including the memory effect and the vertex corrections. The incorporation of the memory effect is through a self consistency procedure that evaluates the order parameter and shear momentum relaxation rates at nonzero frequencies, thereby taking their frequency dependence into account. This approach offers considerable simplification and efficiency in the calculation. The vertex corrections are also demonstrated to have significant effects on the numerical value for the critical viscosity exponent, in contrast to some previous theoretical work which indicated that the vertex corrections tend to cancel out from the final result. By carrying out all of the integrations analytically, we have succeeded in tracing the origin of this discrepancy to an error in earlier work. We provide a thorough treatment of the two-term epsilon expansion, as well as a complete three-dimensional analysis of the fluctuating order-parameter and transverse hydrodynamic modes. The study of the interactions of these modes is carried out to high order so as to arrive at z(eta) = 0.0679+/-0.0007 for comparison with the experimentally observed value, 0.0690+/-0.0006 . PMID- 15783318 TI - Onset of patterns in an oscillated granular layer: continuum and molecular dynamics simulations. AB - We study the onset of patterns in vertically oscillated layers of frictionless dissipative particles. Using both numerical solutions of continuum equations to Navier-Stokes order and molecular dynamics (MD) simulations, we find that standing waves form stripe patterns above a critical acceleration of the cell. Changing the frequency of oscillation of the cell changes the wavelength of the resulting pattern; MD and continuum simulations both yield wavelengths in accord with previous experimental results. The value of the critical acceleration for ordered standing waves is approximately 10% higher in molecular dynamics simulations than in the continuum simulations, and the amplitude of the waves differs significantly between the models. The delay in the onset of order in molecular dynamics simulations and the amplitude of noise below this onset are consistent with the presence of fluctuations which are absent in the continuum theory. The strength of the noise obtained by fit to Swift-Hohenberg theory is orders of magnitude larger than the thermal noise in fluid convection experiments, and is comparable to the noise found in experiments with oscillated granular layers and in recent fluid experiments on fluids near the critical point. Good agreement is found between the mean field value of onset from the Swift-Hohenberg fit and the onset in continuum simulations. Patterns are compared in cells oscillated at two different frequencies in MD; the layer with larger wavelength patterns has less noise than the layer with smaller wavelength patterns. PMID- 15783319 TI - Objective decomposition of the stress tensor in granular flows. AB - A model for the stress tensor in granular flows [Volfson, Tsimring, and Aranson, Phys. Rev. Lett. 90, 254301 (2003)] is correctly generalized to an objective form that is independent of the coordinate system. The objective representation correctly models the isotropic and anisotropic parts of the stress tensor, whereas the original model for stress tensor components is dependent on the coordinate system. This general objective form of the model also relaxes the assumption in the original model that the principal axes of the granular stress tensor be coaxial with that of the "fluid" stress tensor. This generalization expands the applicability of the model to a wider class of granular flows. The objective representation is also useful in analyzing other models based on additive decomposition of the stress tensor in granular flows. PMID- 15783320 TI - Separation of binary granular mixtures under vibration and differential magnetic levitation force. AB - The application of both a strong magnetic field and a magnetic field gradient to a diamagnetic or paramagnetic material can produce a vertical force that acts in concert with the force of gravity. We consider a binary granular mixture in which the two components have different magnetic susceptibilities and therefore experience different effective forces of gravity when subjected to an inhomogeneous magnetic field. Under vertical vibration, such a mixture may rapidly separate into regions almost pure in the two components. We investigate the conditions for this behavior, studying the speed and completeness of separation as a function of differential effective gravity and the frequency and amplitude of vibration. The influence of the cohesive magnetic dipole-dipole interactions on the separation process is also investigated. In our studies insight is gained through the use of a molecular dynamics simulation model. PMID- 15783321 TI - Ejection of dust by elastic waves in collisions between millimeter- and centimeter-sized dust aggregates at 16.5 to 37.5 m/s impact velocities. AB - We report on experiments in which millimeter-sized Si O2 dust aggregates consisting of (sub)-micrometer-sized grains impact into centimeter-sized targets that consist of the same kind of dust particles. The porosity of the granular targets is between 74% and 88%. Impact speeds are between 16.5 and 37.5 m/s with most impacts around 25 m/s . Compaction of the target by the impacting dust aggregate creates a crater which is several millimeters deep and 2-3 cm in diameter. We do not detect a significant amount of ejecta originating at the crater. We do observe a large amount of ejecta though. These are dust granules that are ejected from the whole target surface up to significant distances away from the impact site. This implies that elastic waves induced by the impact are an efficient mechanism to eject material. The estimated mass of these ejecta can be larger than 10 times the projectile mass. The ejecta velocity is uniform across the surface. It is typically 0.5% of the impact velocity. We apply these results to the problem of planetesimal formation. Under microgravity ablation of a dusty body or mass gain in a dust-dust collision might result. This depends on the parameters of the impact. Due to the low ejecta velocities, net growth is also possible in secondary collisions after an eroding primary collision if the body is placed in a gas flow. Thus, for a large number of typical conditions for dust-dust collisions in protoplanetary disks, formation of a larger body results from an impact. PMID- 15783322 TI - Phase diagrams for cohesive particle mixing and segregation. AB - By taking a discrete view of cohesion, we develop a particle-level model which can accurately predict the extent of particle mixing and segregation in cohesive (wet) granular systems. Our model is based on a discrete characterization tool and is used to generate phase diagrams of the predicted particle behavior. These phase diagrams exhibit both mixed and segregated phases where the boundary is determined by the mechanical and surface properties of the particles, such that manipulation of surface properties and/or size/density ratios provides a method to control cohesive particle mixing and segregation. A detailed description of the phase diagram development process as well as quantitative validation of the theoretical results are reported here. PMID- 15783323 TI - Strain softening, yielding, and shear thinning in glassy colloidal suspensions. AB - A microscopic theory for the dependence on external strain, stress, and shear rate of the transient localization length, elastic modulus, alpha relaxation time, shear viscosity, and other dynamic properties of glassy colloidal suspensions is formulated and numerically applied. The approach is built on entropic barrier hopping as the elementary physical process. The concept of an ideal glass transition plays no role, and dynamical slowing down is a continuous, albeit precipitous, process with increasing colloid volume fraction. The relative roles of mechanically driven motion versus thermally activated barrier hopping and transport have been studied. Various scaling behaviors are found for the relaxation time and shear viscosity in both the controlled stress and shear rate mode of rheological experiments. Apparent power law and/or exponential dependences of the elastic modulus and perturbative and absolute yield stresses on colloid volume fraction are predicted. A nonmonotonic dependence of the absolute yield strain on volume fraction is also found. Qualitative and quantitative comparisons of calculations with experiments on high volume fraction glassy colloidal suspensions show encouraging agreement, and multiple testable predictions are made. The theory is generalizable to treat nonlinear rheological phenomena in other soft glassy complex fluids including depletion gels. PMID- 15783324 TI - Kinetics of ergodic-to-nonergodic transitions in charged colloidal suspensions: aging and gelation. AB - There are two types of isotropic disordered nonergodic states in colloidal suspensions: colloidal glasses and gels. In a recent paper [H. Tanaka, J. Meunier, and D. Bonn, Phys. Rev. E 69, 031404 (2004)], we discussed the static aspect of the differences and the similarities between the two. In this paper, we focus on the dynamic aspect. The kinetics of the liquid-glass transition is called "aging," while that of the sol-gel transition is called "gelation." The former is primarily governed by repulsive interactions between particles, while the latter is dominated by attractive interactions. Slowing down of the dynamics during aging reflects the increasing cooperativity required for the escape of a particle from the cage formed by the surrounding particles, while that during gelation reflects the increase in the size of particle clusters towards the percolation transition. Despite these clear differences in the origin of the slowing down of the kinetics between the two, it is not straightforward experimentally to distinguish them in a clear manner. For an understanding of the universal nature of ergodic-to-nonergodic transitions, it is of fundamental importance to elucidate the differences and the similarities in the kinetics between aging and gelation. We consider this problem, taking Laponite suspension as an explicit example. In particular, we focus on the two types of nonergodic states: (i) an attractive gel formed by van der Waals attractions for high ionic strengths and (ii) a repulsive Wigner glass stabilized by long-range Coulomb repulsions for low ionic strengths. We demonstrate that the aging of colloidal Wigner glass crucially differs not only from gelation, but also from the aging of structural and spin glasses. The aging of the colloidal Wigner glass is characterized by the unique cage-forming regime that does not exist in the aging of spin and structural glasses. PMID- 15783325 TI - Light-induced charge separation and storage in titanium oxide gels. AB - We report on the interaction of light with a particular class of media--wet gels, which in contrast to sols of nanoparticles possess a macroscopic bulk structure, and which differ from conventional solids by the existence of the internal liquid solid interface. We show, taking an absorption cross section of trapped electrons from Safrany, Gao, and Rabani [J. Phys. Chem. B 104, 5848 (2000)], that a separation of charges with quantum efficiency as high as 46% appears under the band-gap irradiation of titanium oxide gels: electrons are stored in the gel network as Ti3+ centers, whereas holes are stored in the liquid phase. Under a prolonged UV-laser irradiation, more than 14% of Ti4+ centers can be converted into Ti3+ . Their lifetime can be extremely long and exceeds months at room temperature. The trapped electrons are responsible for a "dark" absorption continuum covering the spectral range from 350 nm to 2.5 mum . PMID- 15783326 TI - Crystallization kinetics of polydisperse colloidal hard spheres. II. Binary mixtures. AB - In this paper we present measurements of the crystallization kinetics of binary mixtures of two different sized hard sphere particles. The growth of the Bragg reflections over time were analyzed to yield the crystallite scattering vector, the total amount of crystal, and the average linear crystal size. It was observed that a particle size distribution skewed to higher sized particles has a less detrimental effect on the crystal structure than a skew to smaller sized particles. In the latter case we observe that initial crystallite growth occurs at only a small number of sites, with further crystallization sites developing at later times. Based on these measurements we elaborate further on the previously proposed growth mechanism whereby crystallization occurs in conjunction with a local fractionation process in the fluid, which significantly affects the kinetic growth of crystallites in polydisperse systems. PMID- 15783327 TI - Structural phase transitions and unusual melting behavior in a classical two dimensional Coulomb bound cluster. AB - The melting properties of a cluster with N equally charged particles confined by a Coulomb potential are studied. The system exhibits a structural phase transition before it melts. The melting process is not dominated by the usual thermal hops between stable states. We also show that the symmetry of the ground state configuration is a dominant factor in determining the melting temperature and that more confined particles in the cluster do not necessarily have a higher melting temperature. PMID- 15783328 TI - Solvent-induced symmetry breaking: varying solvent strength. AB - The triiodide ion is an example of a system where symmetry breaking may be induced by a solvent. The Landau free energy is expected to have a similar form to that for the mean field Ising model, but with solvent strength rather than temperature as the control parameter determining whether there is symmetry breaking. In order to examine the extent of anomalous behavior near the critical point we have studied the properties of the ion in a solvent based on a model for water with charges scaled by a factor lambda . As lambda is increased from zero the solvent strength increases and the system changes from one with no symmetry breaking to one with strong symmetry breaking. The Shannon entropy as a function of lambda shows only a weak maximum near the critical value of lambda= lambda(c) , while the susceptibility has no anomalous behavior. We examine a simple model and show that anomalies near the critical point would increase as the temperature decreases, but divergences associated with a second order phase transition would only be seen in the limit of zero temperature. PMID- 15783329 TI - Griffiths-Wheeler geometrical picture of critical phenomena: experimental testing for liquid-liquid critical points. AB - An experimental approach to the verification of specific relations between thermodynamic properties as predicted from the Griffiths-Wheeler theory of critical phenomena in multicomponent systems is developed for the particular case of ordinary liquid-liquid critical points of binary mixtures. Densities rho(T) , isobaric heat capacities per unit volume C(p)(T) , and previously reported values of the slope of the critical line (dT/dp)c for five critical mixtures are used to check the thermodynamic consistency of C(p) and rho near the critical point. An appropriate treatment of rho (T) data is found to provide the key solution to this issue. In addition, various alternative treatments for C(p)(T) data provide values for both the critical exponent alpha and the ratio between the critical amplitudes of the heat capacity A+/A- that are in agreement with their widely accepted counterparts, whereas two-scale-factor universality is successfully verified in one of the systems studied. PMID- 15783330 TI - Microrheology of cross-linked polyacrylamide networks. AB - Experiments investigating the local viscoelastic properties of a chemically cross linked polymer are performed on polyacrylamide solutions in the sol and the gel regimes using polystyrene beads of varying sizes and surface chemistry as probes. The thermal motions of the probes are measured to obtain the elastic and viscous moduli of the sample. Probe dynamics are measured using two different dynamic light scattering techniques, diffusing wave spectroscopy (DWS) and quasielastic light scattering (QELS) as well as video-based particle tracking. Diffusing wave spectroscopy probes the short-time dynamics of the scatterers while QELS measures the dynamics at larger times. Video-based particle tracking provides a way to investigate the local environment of the individual probe particles. A combination of all the techniques results in a larger range of frequencies that can be probed compared to conventional bulk measurements while providing local information at the level of individual probes. A modified algebraic form of the generalized Stokes-Einstein equation is used to calculate the frequency-dependent moduli. A comparison of microrheological measurements with bulk rheology exhibits striking similarity, confirming the applicability of microrheology for chemically cross-linked polymeric systems. PMID- 15783331 TI - Igniting homogeneous nucleation. AB - Transient homogeneous nucleation is studied in the limit of large critical sizes. Starting from pure monomers, three eras of transient nucleation are characterized in the classic Becker-Doring kinetic equations with two different models of discrete diffusivity: the classic Turnbull-Fisher formula and an expression describing thermally driven growth of the nucleus. The latter diffusivity yields time lags for nucleation which are much closer to values measured in experiments with disilicate glasses. After an initial stage in which the number of monomers decreases, many clusters of small size are produced and a continuous size distribution is created. During the second era, nucleii are increasing steadily in size in such a way that their distribution appears as a wave front advancing towards the critical size for steady nucleation. The nucleation rate at critical size is negligible during this era. After the wave front reaches critical size, it ignites the creation of supercritical clusters at a rate that increases monotonically until its steady value is reached. Analytical formulas for the transient nucleation rate and the time lag are obtained that improve classical ones and compare very well with direct numerical solutions. PMID- 15783332 TI - Sampling-time effects for persistence and survival in step structural fluctuations. AB - The effects of sampling rate and total measurement time have been determined for single-point measurements of step fluctuations within the context of first passage properties. Time dependent scanning tunneling microscopy has been used to evaluate step fluctuations on Ag(111) films grown on mica as a function of temperature (300-410 K) , on screw dislocations on the facets of Pb crystallites at 320 K , and on Al-terminated Si(111) over the temperature range 770-970 K . Although the fundamental time constant for step fluctuations on Ag and Al/Si varies by orders of magnitude over the temperature ranges of measurement, no dependence of the persistence amplitude on temperature is observed. Instead, the persistence probability is found to scale directly with t/delta t where delta t is the time interval used for sampling. Survival probabilities show a more complex scaling dependence, which includes both the sampling interval and the total measurement time t(m) . Scaling with t/delta t occurs only when delta t/ t(m) is a constant. We show that this observation is equivalent to theoretical predictions that the survival probability will scale as delta t/ L(z) , where L is the effective length of a step. This implies that the survival probability for large systems, when measured with fixed values of t(m) or delta t , should also show little or no temperature dependence. PMID- 15783333 TI - Interface unbinding in structured wedges. AB - The unbinding properties of an interface near structured wedges are investigated by discrete models with short range interactions. The calculations demonstrate that interface unbinding takes place in two stages: (i) a continuous filling-like transition in the pure wedge-like parts of the structure, and (ii) a conclusive discontinuous unbinding. In 2D an exact transfer matrix approach allows us to extract the whole interface phase diagram and the precise mechanism at the basis of the phenomenon. The Metropolis Monte Carlo simulations performed in 3D reveal an analogous behavior. The emerging scenario allows us to shed new light onto the problem of wetting of geometrically rough walls. PMID- 15783334 TI - Highly ordered monodomain ionic self-assembled liquid-crystalline materials. AB - Liquid-crystalline properties of the ionic self assembled complex benzenehexacarboxylic- (didodecyltrimethylammonium)6 [BHC- (C12D)6] were investigated by polarizing microscopy, differential scanning calorimetry (DSC), x ray analysis, null ellipsometry, UV and IR spectroscopy. The complex exhibits a bilayer smectic Sm- A2 liquid-crystalline phase and aligns spontaneously. Alignment properties do not depend on the hydrophobic or hydrophilic treatment of the surfaces. The aligned complex possesses a negative (delta n=-0.02) homeotropically oriented optical axis, with layers aligned parallel to the surface. X-ray analysis of the aligned sample revealed a lamellar structure with a d spacing of 3.15 nm, consisting of sublayers of thicknesses d1 = 1.41 and d2 = 1.74 nm . This was confirmed by simple geometrical calculations and detailed temperature-dependent investigations, revealing that the first layer contains the BHC molecules and oppositely charged groups of the surfactants, and the second the alkyl tails of the surfactant. Changes in the order parameters (as calculated from the IR investigations) are correlated with the phase transitions as found by DSC. The properties of the complex are strongly influenced by the ionic interactions within the complex. The presence of these groups slows down the dynamics within the material sufficiently to allow for crystallization of the complex from an aligned LC phase into a single crystal domain, as well as restricting the transition to the isotropic phase. PMID- 15783335 TI - Effects of intramolecular dipolar coupling on the isotropic-nematic phase transition of a hard spherocylinder fluid. AB - The thermodynamics of a simple model, containing the minimum set of features required to provide liquid crystal-like phase behavior and the dipolar coupling observable in the NMR spectrum of orientationally ordered fluids, are presented within the framework of Onsager theory. The model comprises a fluid of hard spherocylinders with a pair of embedded freely rotating magnetic dipoles. The behavior of the isotropic-nematic phase transition is explored as a function magnetic field strength and of the relative orientation between the nematic director and the external magnetic field. When the field and director are aligned the phase diagram is similar to those predicted for a hard rod fluid in flow fields, electric fields, and magnetic fields, with the field promoting orientational order in the fluid and the isotropic-nematic phase transition being replaced by a paranematic-nematic phase transition. In contrast, when the field and director are perpendicular, the field destabilizes the nematic phase and the phase transition is shifted to higher densities. The variation of the mean magnetic moment and the dipolar coupling are examined as a function of the orientational structure of the fluid. The model is used to support the hypothesis that dipolar couplings observed in the spectra of human leg muscle originate from nematic-like liquid crystal phases in relatively small metabolite molecules. The fitted theoretical predictions of the dependence of the dipolar coupling on the orientation of the field with respect to the nematic director are shown to provide a good description of the experimental data. PMID- 15783336 TI - Shear flow in nematic liquid crystals: Freedericksz transition as a bifurcation. AB - We numerically investigate the Freedericksz transition for steady state plane shear flow of nematic liquid crystals between two parallel plates in the presence of external magnetic fields. Three typical configurations with the external field in the plane of the flow and perpendicular to it, in the plane and along the flow, and where it is perpendicular to the plane of the flow are considered. In each case, the Freedericksz transition is studied as a bifurcation problem. Beginning with a steady state shear flow, solutions corresponding to slowly increasing magnetic fields and those corresponding to fields which are suddenly turned on at a given intensity are studied. For a typical idealized nematic, we show that the symmetric pitch-fork bifurcation in the absence of shear becomes a transcritical bifurcation from the trivial solution in one configuration while in another it resembles a disconnected pitch-fork where the turning point of the disconnected branch is a generic singularity in the absence of symmetry or a trivial solution. PMID- 15783337 TI - Effect of high spontaneous polarization on defect structures and orientational dynamics of tilted chiral smectic freely suspended films. AB - The director structure around topological defects and in 2pi walls in the two dimensional orientation field of thin freely suspended films of tilted chiral smectic liquid crystal is observed to minimize splay of the spontaneous polarization. Concentric ring patterns in the director field unwind more slowly in higher polarization films. These experiments confirm that polarization space charge increases the effective elasticity of static polarization-splay distortions and that it attracts ionic charge, leading to an increase in the effective orientational viscosity of the director field. PMID- 15783338 TI - Using particle shape to induce tilted and bistable liquid crystal anchoring. AB - We use Monte Carlo simulations of hard Gaussian overlap (HGO) particles symmetrically confined in slab geometry to investigate the role of particle substrate interactions on liquid crystalline anchoring. Despite the restriction here to purely steric interactions and smooth substrates, a range of behaviors are captured, including tilted anchoring and homeotropic-planar bistability. These macroscopic behaviors are all achieved through appropriate tuning of the microscopics of the HGO-substrate interaction, based upon nonadditive descriptions for the HGO-substrate shape parameter. PMID- 15783339 TI - Induced and spontaneous deracemization in bent-core liquid crystal phases and in other phases doped with bent-core molecules. AB - Recently discovered chiral properties of several bent-core smectic liquid crystal phases are summarized and discussed in detail under the assumption that typical bent-core molecules may exist in chiral conformational states and are achiral only on average. Results of atomistic computer simulations are presented which indicate the existence of strongly chiral conformational states for typical bent core mesogens. A theory is developed which describes a possible shift of equilibrium between left- and right-handed conformations in a macroscopically chiral phase. The theory describes a chirality induction in the B2 bent-core phase and a reduction of the helical pitch in cholesteric and chiral SmC* phases doped with bent-core molecules. Finally, the possibility of spontaneous deracemization in bent-core smectic phases is discussed in detail. PMID- 15783340 TI - Routes to spatiotemporal chaos in the rheology of nematogenic fluids. AB - With a view to understanding the "rheochaos" observed in recent experiments in a variety of orientable fluids, we study numerically the equations of motion of the spatiotemporal evolution of the traceless symmetric order parameter of a sheared nematogenic fluid. In particular we establish, by decisive numerical tests, that the irregular oscillatory behavior seen in a region of parameter space where the nematic is not stably flow-aligning is in fact spatiotemporal chaos. We outline the dynamical phase diagram of the model and study the route to the chaotic state. We find that spatiotemporal chaos in this system sets in via a regime of spatiotemporal intermittency, with a power-law distribution of the widths of laminar regions, as in H. Chate and P. Manneville, Phys. Rev. Lett. 58, 112 (1987). Further, the evolution of the histogram of band sizes shows a growing length scale as one moves from the chaotic towards the flow-aligned phase. Finally we suggest possible experiments in which one can observe the intriguing behaviors discussed here. PMID- 15783341 TI - Elasticity of smectic-A elastomers. AB - We present a fully nonlinear model of the elasticity smectic-A elastomers, and compare our results with a wide range of experimental observations: extreme Poisson ratios, the in-plane modulus, the modulus before and after threshold to layer rotation in response to stretches along the layer normal, the threshold strain, the characteristic, and singular rotation of layers after the threshold. We calculate the x-ray scattering from rotating layers and compare with available data. The model is derived in two ways: from geometrical constraints imposed by layers on a nematic elastomer, and from application of statistical mechanics to a microscopic model of the effect of crosslink points confined in a corrugated potential. PMID- 15783342 TI - Light-induced changes of optical and electrical properties in bent-core azo compounds. AB - We have studied the optical and electrical properties of two bent-core substances with an azo linkage in their cores. Pump-probe laser studies, direct textural observations, and spectrophotometric recordings show an initial decrease of light transmission, which at larger light intensities (approximately 1 mW/mm2) is followed by a bleaching. Simultaneously the electrical properties (electric conductivity, antiferroelectric polarization, switching threshold, and switching time) decreased monotonically with increasing light intensities. The monotonic decrease of electrical properties indicates that the darkening and bleaching have the same origin, namely, the photochemical isomerization of the azo linkage from the trans to the cis isomer. The material with cis isomer has a lower clearing point and phase separates from the trans-rich domains. Initially the size of the separated isotropic domains is below the visible range, which causes increased scattering. As the size of the isotropic domains increase the scattering disappears and the transmittance becomes the average of the transmittances in the polar tilted smectic and isotropic phases. PMID- 15783343 TI - Theory of the dielectric susceptibility of nonpolar biaxial liquid crystals. AB - A statistical theory of the dielectric susceptibility of nonpolar liquid crystals, whose constituent molecules are biaxial, is proposed. The ordering is described by means of the mean field theory, in which a set of basic functions is introduced. The dielectric susceptibilities are derived using a generalized Clausius-Mossotti approach. The theory was used to calculate the temperature and the density dependence of the order parameters and of the susceptibilities. On increasing the density, an ordering reversal of the corresponding susceptibilities is obtained. PMID- 15783344 TI - Two kinds of smectic-C(alpha)* subphases in a liquid crystal and their relative stability dependent on the enantiomeric excess as elucidated by electric-field induced birefringence experiment. AB - The electric-field-induced birefringence has been investigated by using a photoelastic modulator, with a view to obtaining a molecular model for the subphases produced by the frustration between ferroelectricity and antiferroelectricity in the chiral smectic liquid crystals. It has been found that even in the bulk, there exist two subphases in the smectic-C(alpha)* (Sm C(alpha)*) temperature range. By extending the Emelyanenko-Osipov model [Phys. Rev. E 68, 051703 (2003)] to include the temperature dependence of the tilt angle, we have alluded to a possible lifting of the degeneracy at the frustration point P(alpha) , where Sm-C(A)*, Sm-C*, and Sm-A have the same free energy. This leads to the appearance of uniaxial Sm-C(alpha)* characterized by short-pitch helical structures and consequently with a pitch much lower than the optical wavelength. The numerical calculations indicate that the short pitch may generally increase or decrease monotonically with temperature. Depending on the parameter value that represents the relative strength of ferroelectricity and antiferroelectricity, the short-pitch temperature variation may abruptly change from increase to decrease at a temperature; this can be assigned to the observed phase transition between the two Sm-C(alpha)* subphases. PMID- 15783345 TI - Constrained dynamics of a polymer ring enclosing a constant area. AB - The dynamics of an ideal polymer ring enclosing a constant algebraic area is studied. The constraint of a constant area is found to couple the dynamics of the two Cartesian components of the position vector of the polymer ring through the Lagrange multiplier function which is time dependent. The time dependence of the Lagrange multiplier is evaluated in a closed form both at short and long times. At long times, the time dependence is weak, and is mainly governed by the inverse of the first mode of the area. The presence of the constraint changes the nature of the relaxation of the internal modes. The time correlation of the position vectors of the ring is found to be dominated by the first Rouse mode which does not relax even at very long times. The mean square displacement of the radius vector is found to be diffusive, which is associated with the rotational diffusion of the ring. PMID- 15783346 TI - Topographical maps as complex networks. AB - The neuronal networks in the mammalian cortex are characterized by the coexistence of hierarchy, modularity, short and long range interactions, spatial correlations, and topographical connections. Particularly interesting, the latter type of organization implies special demands on developing systems in order to achieve precise maps preserving spatial adjacencies, even at the expense of isometry. Although the object of intensive biological research, the elucidation of the main anatomic-functional purposes of the ubiquitous topographical connections in the mammalian brain remains an elusive issue. The present work reports on how recent results from complex network formalism can be used to quantify and model the effect of topographical connections between neuronal cells over the connectivity of the network. While the topographical mapping between two cortical modules is achieved by connecting nearest cells from each module, four kinds of network models are adopted for implementing intramodular connections, including random, preferential-attachment, short-range, and long-range networks. It is shown that, though spatially uniform and simple, topographical connections between modules can lead to major changes in the network properties in some specific cases, depending on intramodular connections schemes, fostering more effective intercommunication between the involved neuronal cells and modules. The possible implications of such effects on cortical operation are discussed. PMID- 15783347 TI - Detection of pulses in a colored noise setting. AB - Cortical neurons are exposed to a considerable amount of synaptic background activity, which increases the neurons' conductance and which leads to a fluctuating membrane potential. Here we investigate how the presence and the properties of this background noise influence the ability of a neuron to detect transient inputs, a task that is important for coincidence detection as well as for the detection of synchronous spiking events in a neural system. Using a leaky integrate-and-fire neuron as well as a biologically more realistic Hodgkin-Huxley type point neuron we find that noise enhances the detection of subthreshold input pulses and that the phenomenon of stochastic resonance occurs. When the noise is colored, pulse detection becomes more robust, because the number of false positive events decreases with increasing temporal correlation while the number of correctly detected events is almost unaffected. Therefore, the optimal variance of the noise also changes with the degree of temporal correlations of the background activity. For the integrate-and-fire model these effects can be described using an ansatz by Brunel and Sergi [J. Theor. Biol. 195, 87 (1998)]. Numerical simulations show that the leaky integrate-and-fire model and the Hodgkin-Huxley type point neuron behave qualitatively similarly. PMID- 15783348 TI - Coupled dynamics of voltage and calcium in paced cardiac cells. AB - We investigate numerically and analytically the coupled dynamics of transmembrane voltage and intracellular calcium cycling in paced cardiac cells using a detailed physiological model, and its reduction to a three-dimensional discrete map. The results provide a theoretical framework to interpret various experimentally observed modes of instability ranging from electromechanically concordant and discordant alternans to quasiperiodic oscillations of voltage and calcium. PMID- 15783349 TI - Theoretical studies of the kinetics of mechanical unfolding of cross-linked polymer chains and their implications for single-molecule pulling experiments. AB - We have used kinetic Monte Carlo simulations to study the kinetics of unfolding of cross-linked polymer chains under mechanical loading. As the ends of a chain are pulled apart, the force transmitted by each cross-link increases until it ruptures. The stochastic cross-link rupture process is assumed to be governed by first order kinetics with a rate that depends exponentially on the transmitted force. We have performed random searches to identify optimal cross-link configurations whose unfolding requires a large applied force (measure of strength) and/or large dissipated energy (measure of toughness). We found that such optimal chains always involve cross-links arranged to form parallel strands. The location of those optimal strands generally depends on the loading rate. Optimal chains with a small number of cross-links were found to be almost as strong and tough as optimal chains with a large number of cross-links. Furthermore, optimality of chains with a small number of cross-links can be easily destroyed by adding cross-links at random. The present findings are relevant for the interpretation of single molecule force probe spectroscopy studies of the mechanical unfolding of "load-bearing" proteins, whose native topology often involves parallel strand arrangements similar to the optimal configurations identified in the study. PMID- 15783350 TI - Viscoelastic dynamics of spherical composite vesicles. AB - A micromechanical model for the low-frequency dynamics of spherical composite vesicles (CVs) is proposed. Solid-like viscoelastic properties of the CVs are taken into account. The equations of motion of a CV surrounded by a viscous liquid are derived. They have discrete solutions which describe linearly coupled stretching and bending relaxation modes and an independent shear mode. The qualitative difference between the bending modes excited in a spherical vesicle and that in a flat membrane is demonstrated. The shear elasticity of the CVs gives an essential contribution to the relaxation rate of the bending mode at small wave numbers. It is also shown that even in an incompressible spherical vesicle with a finite shear modulus, the bending mode involves both radial and tangent displacements. These reasons make both in-plane and out-of-plane low frequency responses of the CV quite different with respect to those of the flat membrane. To compare our theoretical results with published experimental data, the power spectra of the actin-coated CV are calculated. PMID- 15783351 TI - Multiscale entropy analysis of biological signals. AB - Traditional approaches to measuring the complexity of biological signals fail to account for the multiple time scales inherent in such time series. These algorithms have yielded contradictory findings when applied to real-world datasets obtained in health and disease states. We describe in detail the basis and implementation of the multiscale entropy (MSE) method. We extend and elaborate previous findings showing its applicability to the fluctuations of the human heartbeat under physiologic and pathologic conditions. The method consistently indicates a loss of complexity with aging, with an erratic cardiac arrhythmia (atrial fibrillation), and with a life-threatening syndrome (congestive heart failure). Further, these different conditions have distinct MSE curve profiles, suggesting diagnostic uses. The results support a general "complexity-loss" theory of aging and disease. We also apply the method to the analysis of coding and noncoding DNA sequences and find that the latter have higher multiscale entropy, consistent with the emerging view that so-called "junk DNA" sequences contain important biological information. PMID- 15783352 TI - Three-dimensional bead position histograms reveal single-molecule nanomechanics. AB - We describe a method to investigate the structure and elasticity of macromolecules by a combination of single molecule experiments and kinematic modeling. With a photonic force microscope, we recorded spatial position histograms of a fluctuating microsphere tethered to full-length myosin-II. Assuming only that the molecule consists of concatenated rigid segments, a model derived from robot kinematics allows us to relate these histograms to the molecule's segment lengths and bending stiffnesses. Both our calculated position distributions and the experimental data show an asymmetry characteristic of a mixed entropic-enthalpic spring. Our model that fits best to experimental line profiles has two intramolecular hinges, one at the bound head domain, and another about 50 nm down the myosin tail, with a summed bending stiffness of about 3 k(B)T/rad. PMID- 15783353 TI - Approximate solution to the bidomain equations for defibrillation problems. AB - The bidomain model can be used for calculating the electrical potential in the heart during defibrillation. However, this model consists of a coupled system of two partial differential equations that are, in general, difficult and time consuming to solve. In this paper, we present an approximate, iterative method of solving the bidomain equations. After working out the general method, we apply it to four problems: (i) a cylindrical strand in a uniform electric field, (ii) a nonuniform electric field applied to tissue with straight fibers, (iii) a spherical heart in a uniform electric field, and (iv) a two-dimensional sheet of cardiac tissue with curving fibers. Finally, we analyze the general case of three dimensions. PMID- 15783354 TI - Exact theory of kinkable elastic polymers. AB - The importance of nonlinearities in material constitutive relations has long been appreciated in the continuum mechanics of macroscopic rods. Although the moment (torque) response to bending is almost universally linear for small deflection angles, many rod systems exhibit a high-curvature softening. The signature behavior of these rod systems is a kinking transition in which the bending is localized. Recent DNA cyclization experiments by Cloutier and Widom have offered evidence that the linear-elastic bending theory fails to describe the high curvature mechanics of DNA. Motivated by this recent experimental work, we develop a simple and exact theory of the statistical mechanics of linear-elastic polymer chains that can undergo a kinking transition. We characterize the kinking behavior with a single parameter and show that the resulting theory reproduces both the low-curvature linear-elastic behavior which is already well described by the worm-like chain model, as well as the high-curvature softening observed in recent cyclization experiments. PMID- 15783355 TI - Nucleotide correlations and electronic transport of DNA sequences. AB - We use a tight-binding formulation to investigate the transmissivity and wave packet dynamics of sequences of single-strand DNA molecules made up from the nucleotides guanine G , adenine A , cytosine C , and thymine T . In order to reveal the relevance of the underlying correlations in the nucleotides distribution, we compare the results for the genomic DNA sequence with those of two artificial sequences: (i) the Rudin-Shapiro one, which has long-range correlations; (ii) a random sequence, which is a kind of prototype of a short range correlated system, presented here with the same first-neighbor pair correlations of the human DNA sequence. We found that the long-range character of the correlations is important to the persistence of resonances of finite segments. On the other hand, the wave-packet dynamics seems to be mostly influenced by the short-range correlations. PMID- 15783356 TI - Formation of loops in DNA under tension. AB - We study the formation of loops along a DNA molecule under applied tension, as might occur in single-DNA micromanipulation experiments with proteins which are able to simultaneously bind two DNA sites. We consider the case of "bare" DNA in the loop, which forms a "teardrop" shape, and the case where a single DNA-bending protein produces a "kink" in the middle of the loop; the presence of a right angle kink in the loop reduces its bending energy by a factor of 3. Using the bending energy plus an estimate of the free energies associated with fluctuations and the elasticity of the extended nonlooped DNA, we obtain a probability distribution for loops as a function of loop size and force. Force strongly suppresses formation of all loops, but suppresses large loops more severely than small ones. This quenching effect of force is reduced in the presence of a kink in the loop. We also calculate the speed at which length is absorbed into loops between arbitrary positions along the DNA (i.e., for non-sequence-specific loop forming proteins). The speed of retraction of the molecule decays as a stretched exponential function of the force with characteristic force scales depending on the geometry of the loops. PMID- 15783357 TI - Non-Michaelis-Menten kinetics model for conductance of low-conductance potassium ion channels. AB - A reduced kinetics model is proposed for ion permeation in low-conductance potassium ion channels with zero net electrical charge in the selectivity filter region. The selectivity filter is assumed to be the only conductance-determining part of the channel. Ion entry and exit rate constants depend on the occupancy of the filter due to ion-ion interactions. The corresponding rates are assumed slow relative to the rates of ion motion between binding sites inside the filter, allowing a reduction of the kinetics model of the filter by averaging the entry and exit rate constants over the states with a particular occupancy number. The reduced kinetics model for low-conductance channels is described by only three states and two sets of effective rate constants characterizing transitions between these states. An explicit expression for the channel conductance as a function of symmetrical external ion concentration is derived under the assumption that the average electrical mobility of ions in the selectivity filter region in a limited range of ion concentrations does not depend on these concentrations. The simplified conductance model is shown to provide a good description of the experimentally observed conductance-concentration curve for the low-conductance potassium channel Kir2.1, and also predicts the mean occupancy of the selectivity filter of this channel. We find that at physiological external ion concentrations this occupancy is much lower than the value of two ions observed for one of the high-conductance potassium channels, KcsA. PMID- 15783358 TI - Dependence of RNA secondary structure on the energy model. AB - We analyze a microscopic RNA model, which includes two widely used models as limiting cases; namely, it contains terms for bond as well as for stacking energies. We numerically investigate possible changes in the qualitative and quantitative behavior while going from one model to the other; in particular, we test whether a transition occurs when continuously moving from one model to the other. For this we calculate various thermodynamic quantities, at both zero temperature and finite temperatures. All calculations can be done efficiently in polynomial time by a dynamic programming algorithm. We do not find a sign for the transition between the models, but the critical exponent nu of the correlation length, describing the phase transition in all models to an ordered low temperature phase, seems to depend continuously on the model. Finally, we apply the epsilon -coupling method to study low-energy excitations. The exponent theta describing the energy scaling of the excitations seems to depend not much on the energy model. PMID- 15783359 TI - Deformation of a helical filament by flow and electric or magnetic fields. AB - Motivated by recent advances in the real-time imaging of fluorescent flagellar filaments in living bacteria [Turner, Ryu, and Berg, J. Bacteriol. 82, 2793 (2000)], we compute the deformation of a helical elastic filament due to flow and external magnetic or high-frequency electric fields. Two cases of deformation due to hydrodynamic drag are considered: the compression of a filament rotated by a stationary motor and the extension of a stationary filament due to flow along the helical axis. We use Kirchhoff rod theory for the filament, and work to linear order in the deflection. Hydrodynamic forces are described first by resistive force theory, and then for comparison by the more accurate slender-body theory. For helices with a short pitch, the deflection in axial flow predicted by slender body theory is significantly smaller than that computed with resistive-force theory. Therefore, our estimate of the bending stiffness of a flagellar filament is smaller than that of previous workers. In our calculation of the deformation of a polarizable helix in an external field, we show that the problem is equivalent to the classical case of a helix deformed by forces applied only at the ends. PMID- 15783360 TI - Interactions between water soluble porphyrin-based star polymer and amino acids: spectroscopic evidence of molecular binding. AB - Molecular interactions giving rise to stable complexes between an uncharged water soluble cobalt-porphyrin and amino acids are investigated by time-resolved fluorescence, uv-vis, and circular dichroism measurements. This metalloporphyrin seems to act, by means of the coordination site of the cobalt of the core, as a recognition host, preferentially, with amino acids possessing aromatic groups. The binding with aliphatic amino acids requires longer time scales to be efficient and likely involves a slow kinetic process. The experimental findings suggest that, besides the metal(host)-N(guest) coordination bond, which is the common requisite for all amino acids, a preferential interaction with aromatic groups exists there. The solubility in water of the molecule, guaranteed by the polyethylene glycol arms as peripheral substituents, in the absence of electric charges, allows for a more selective discrimination of the binding process with respect to other water-soluble charged porphyrins. The interest devoted to the porphyrin-based star polymer and its recognition properties is, therefore, founded on the potential use either in polymeric matrices for material science or in aqueous solution for bioscience. PMID- 15783361 TI - Modulated structures in nematic monolayers formed by symmetric molecules. AB - An analysis based on symmetry yields a general form for the deformation elastic energy of a nematic monolayer, formed by achiral symmetric molecules, deposited on a solid substrate. Lifshitz-invariant-like terms in the energy, which originate from the substrate field, can induce a modulated-tilt state if the anchoring energy is sufficiently low. A way to enhance the symmetry breaking is to apply a destabilizing magnetic or electric field that serves to lower the anchoring energy. In the case of an initial state with homeotropic alignment, the phase diagram displays a cusp-shaped tilt-modulated state intervening between two uniform tilt states. PMID- 15783362 TI - Ion conduction in the KcsA potassium channel analyzed with a minimal kinetic model. AB - We use a model by Nelson to study the current-voltage and conductance concentration curves of bacterial potassium channel KcsA without assuming rapid ion translocation. Ion association to the channel filter is rate controlling at low concentrations, but dissociation and transport in the filter can limit conduction at high concentration for ions other than K+. The absolute values of the effective rate constants are tentative but the relative changes in these constants needed to qualitatively explain the experiments should be of significance. PMID- 15783363 TI - Non-Gaussian fluctuations arising from finite populations: Exact results for the evolutionary Moran process. AB - The appropriate description of fluctuations within the framework of evolutionary game theory is a fundamental unsolved problem in the case of finite populations. The Moran process recently introduced into this context in Nowak, [Nature (London) 428, 646 (2004)] defines a promising standard model of evolutionary game theory in finite populations for which analytical results are accessible. In this paper, we derive the stationary distribution of the Moran process population dynamics for arbitrary 2 x 2 games for the finite-size case. We show that a nonvanishing background fitness can be transformed to the vanishing case by rescaling the payoff matrix. In contrast to the common approach to mimic finite size fluctuations by Gaussian distributed noise, the finite-size fluctuations can deviate significantly from a Gaussian distribution. PMID- 15783364 TI - Dynamical generalization of nonequilibrium work relation. AB - The evolution of systems in contact with thermal, chaotic, or turbulent surroundings-often modeled with stochastic equations of motion-can be particularly complex when these equations of motion are nonautonomous, that is, when external parameters of the surroundings are varied with time. In this paper we establish a rigorous equality relating the nonautonomous behavior of such a system, to solutions of the corresponding autonomous equations of motion, for arbitrary initial conditions. If the system is initially in thermal equilibrium, we recover previously known results relating nonequilibrium work values to equilibrium probability distributions. We discuss specific examples of our result, and suggest an experimental setting in which it might be verified. PMID- 15783365 TI - Inhomogeneous evolution of subgraphs and cycles in complex networks. AB - Subgraphs and cycles are often used to characterize the local properties of complex networks. Here we show that the subgraph structure of real networks is highly time dependent: as the network grows, the density of some subgraphs remains unchanged, while the density of others increase at a rate that is determined by the network's degree distribution and clustering properties. This inhomogeneous evolution process, supported by direct measurements on several real networks, leads to systematic shifts in the overall subgraph spectrum and to an inevitable overrepresentation of some subgraphs and cycles. PMID- 15783366 TI - Power-law correlated processes with asymmetric distributions. AB - Motivated by the fact that many physical systems display (i) power-law correlations together with (ii) an asymmetry in the probability distribution, we propose a stochastic process that can model both properties. The process depends on only two parameters, where one controls the scaling exponent of the power-law correlations, and the other controls the degree of asymmetry in the distributions leaving the correlations unaffected. We apply the process to air humidity data and find that the statistical properties of the process are in a good agreement with those observed in the data. PMID- 15783367 TI - Defect-enhanced anomaly in frequency synchronization of asymmetrically coupled spatially extended systems. AB - We analytically establish and numerically show that anomalous frequency synchronization occurs in a pair of asymmetrically coupled chaotic space extended oscillators. The transition to anomalous behaviors is crucially dependent on asymmetries in the coupling configuration, while the presence of phase defects has the effect of enhancing the anomaly in frequency synchronization with respect to the case of merely time chaotic oscillators. PMID- 15783368 TI - Fluctuation formula in the Nose-Hoover thermostated Lorentz gas. AB - In this paper we examine numerically the Gallavotti-Cohen fluctuation formula for phase-space contraction rate and entropy production rate fluctuations in the Nose Hoover thermostated periodic Lorentz gas. Our results indicate that while the phase-space contraction rate fluctuations violate the fluctuation formula near equilibrium states, the entropy production rate fluctuations obey this formula near and far from equilibrium states as well. PMID- 15783369 TI - Critical exponents of Nikolaevskii turbulence. AB - We study the spatial power spectra of Nikolaevskii turbulence in one-dimensional space. First, we show that the energy distribution in wave-number space is extensive in nature. Then, we demonstrate that, when varying a particular parameter, the spectrum becomes qualitatively indistinguishable from that of Kuramoto-Sivashinsky turbulence. Next, we derive the critical exponents of turbulent fluctuations. Finally, we argue that in some previous studies, parameter values for which this type of turbulence does not appear were mistakenly considered, and we resolve inconsistencies obtained in previous studies. PMID- 15783370 TI - Laser-driven hard-x-ray generation based on ultrafast selected energy X-ray absorption spectroscopy measurements of Ni compounds. AB - Three Ni compounds were studied by ultrafast selected energy x-ray absorption spectroscopy using a laser-driven electron x-ray source with a tungsten target. The measured K edges of these Ni compounds using this self-referencing method were made identical to those measured with synchrotron x-ray sources. This enabled us to determine the absolute peak positions of tungsten Lalpha(1) and Lalpha(2) emitted from this source to be within 1 eV of those from the neutral tungsten atoms, which strongly suggested that the x rays were emitted from high energy electrons interacting with tungsten atoms in the solid target. This is the best evidence to date that directly supports the cold atom x-ray generation theory. PMID- 15783371 TI - Phase twisted modes and current reversals in a lattice model of waveguide arrays with nonlinear coupling. AB - We consider a lattice model for waveguide arrays embedded in nonlinear Kerr media. Inclusion of nonlinear coupling results in many phenomena involving complex, phase-twisted, stationary modes. The norm (Poynting power) current of stable plane-wave solutions can be controlled in magnitude and direction, and may be reversed without symmetry-breaking perturbations. Also stable localized phase twisted modes with zero current exist, which for particular parameter values may be compact and expressed analytically. The model also describes coupled Bose Einstein condensates. PMID- 15783372 TI - Joint probability distributions for a class of non-Markovian processes. AB - We consider joint probability distributions for the class of coupled Langevin equations introduced by Fogedby [H. C. Fogedby, Phys. Rev. E 50, 1657 (1994)]. We generalize well-known results for the single-time probability distributions to the case of N -time joint probability distributions. It is shown that these probability distribution functions can be obtained by an integral transform from distributions of a Markovian process. The integral kernel obeys a partial differential equation with fractional time derivatives reflecting the non Markovian character of the process. PMID- 15783373 TI - Statistical physics of grain-boundary engineering. AB - Percolation theory is now standard in the analysis of polycrystalline materials where the grain boundaries can be divided into two distinct classes, namely "good" boundaries that have favorable properties and "bad" boundaries that seriously degrade the material performance. Grain-boundary engineering (GBE) strives to improve material behavior by engineering the volume fraction c and arrangement of good grain boundaries. Two key percolative processes in GBE materials are the onset of percolation of a strongly connected aggregate of grains, and the onset of a connected path of weak grain boundaries. Using realistic polycrystalline microstructures, we find that in two dimensions the threshold for strong aggregate percolation c(SAP) and the threshold for weak boundary percolation c(WBP) are equivalent and have the value c(SAP) = c(WBP) =0.38 (1) , which is slightly higher than the threshold found for regular hexagonal grain structures, c(RH) =2 sin (pi/18) =0.347... . In three dimensions strong aggregate percolation and weak boundary percolation occur at different locations and we find c(SAP) =0.12 (3) and c(WBP) =0.77 (3) . The critical current in high T(c) materials and the cohesive energy in structural systems are related to the critical manifold problem in statistical physics. We develop a theory of critical manifolds in GBE materials, which has three distinct regimes: (i) low concentrations, where random manifold theory applies, (ii) critical concentrations where percolative scaling theory applies, and (iii) high concentrations, c> c(SAP) , where the theory of periodic elastic media applies. Regime (iii) is perhaps most important practically and is characterized by a critical length L(c) , which is the size of cleavage regions on the critical manifold. In the limit of high contrast epsilon-->0 , we find that in two dimensions L(c) proportional, gc/ (1-c) , while in three dimensions L(c) proportional, g exp [ b(0) c/ (1-c) ] / [c (1-c) ](1/2) , where g is the average grain size, epsilon is the ratio of the bonding energy of the weak boundaries to that of the strong boundaries, and b(0) is a constant which is of order 1. Many of the properties of GBE materials can be related to L(c) , which diverges algebraically on approach to c=1 in two dimensions, but diverges exponentially in that limit in three dimensions. We emphasize that GBE percolation processes and critical manifold behavior are very different in two dimensions as compared to three dimensions. For this reason, the use of two dimensional models to understand the behavior of bulk GBE materials can be misleading. PMID- 15783374 TI - Weight-driven growing networks. AB - We study growing networks in which each link carries a certain weight (randomly assigned at birth and fixed thereafter). The weight of a node is defined as the sum of the weights of the links attached to the node, and the network grows via the simplest weight-driven rule: A newly added node is connected to an already existing node with the probability which is proportional to the weight of that node. We show that the node weight distribution n (w) has a universal tail, that is, it is independent of the link weight distribution: n (w) approximately w(-3) as w-->infinity . Results are particularly neat for the exponential link weight distribution when n (w) is algebraic over the entire weight range. PMID- 15783375 TI - Dynamic phases in the two-dimensional underdamped driven Frenkel-Kontorova model. AB - We study the nonlinear dc response of a two-dimensional underdamped system of interacting atoms subject to an isotropic periodic external potential with triangular symmetry. When driving force increases, the system transfers from a disorder locked state to an ordered sliding state corresponding to a moving crystal. By varying the values of the effective elastic constant, damping, and temperature, we found different scenarios and intermediate phases during the ordering transition. For a soft atomic layer, the system passes through a plastic channel regime that appears as a steady-state regime at higher values of the damping coefficient. For high values of the effective elastic constant, when the atomic layer is stiff, the intermediate plastic phase corresponds to a traffic jam regime with immobile islands in the sea of running atoms. At a high driving of the stiff layer, a solitonlike elastic flow of atoms has been observed. PMID- 15783376 TI - Hysteresis and magnetization jumps in the T=0 dynamics of spin glasses. AB - We present results from Monte Carlo simulations of hysteresis in the zero temperature ( T=0 ) dynamics of the Sherrington-Kirkpatrick spin glass model. We study the statistics of magnetization-jumps (denoted as Deltam ) in response to a time-dependent magnetic field H (t) , which increases or decreases with constant increments Delta as H(t)-->H(t)+/-Delta . In particular, we focus on the field dependence of the Deltam -distribution function P(Deltam,H) . We formulate arguments to understand the variation of P ( Deltam,H ) along the hysteresis loop in the weak-disorder limit. PMID- 15783377 TI - Crack roughness and avalanche precursors in the random fuse model. AB - We analyze the scaling of the crack roughness and of avalanche precursors in the two-dimensional random fuse model by numerical simulations, employing large system sizes and extensive sample averaging. We find that the crack roughness exhibits anomalous scaling, as recently observed in experiments. The roughness exponents (zeta, zeta(loc) ) and the global width distributions are found to be universal with respect to the lattice geometry. Failure is preceded by avalanche precursors whose distribution follows a power law up to a cutoff size. While the characteristic avalanche size scales as s(0) approximately L(D) , with a universal fractal dimension D , the distribution exponent tau differs slightly for triangular and diamond lattices and, in both cases, it is larger than the mean-field (fiber bundle) value tau=5/2 . PMID- 15783379 TI - Group-based Yule model for bipartite author-paper networks. AB - This paper presents a model for author-paper networks, which is based on the assumption that authors are organized into groups and that, for each research topic, the number of papers published by a group is based on a success-breeds success model. Collaboration between groups is modeled as random invitations from a group to an outside member. To analyze the model, a number of different metrics that can be obtained in author-paper networks were extracted. A simulation example shows that this model can effectively mimic the behavior of a real-world author-paper network, extracted from a collection of 900 journal papers in the field of complex networks. PMID- 15783378 TI - Evolution kinetics and phase transitions of complex adaptive systems. AB - Motivated by the fact that a population of competing agents never set up a true stationary distribution, we propose a theory of evolution kinetics for complex adaptive systems. The formula derived for the survival probability is used to describe different phases in the population evolution when the prize-to-fine ratio as well as time changes. A kinetic phase diagram is obtained to show the phase boundaries for the self-segregating region, the intermediate region, and the clustering region. The kinetic evolutionary equations of strategy distribution are also established and used to give the root-mean-square separation of strategy distribution. It is revealed that the rootstock of the phase transitions is ascribed to the cooperation and competition among agents with different gene values for a limited resource. PMID- 15783380 TI - Edge phase transitions of the tricritical Potts model in two dimensions. AB - Using Monte Carlo techniques and finite-size analysis, we investigate several two dimensional lattice models with open edges, including the Blume-Capel model and the q=1 and 3 Potts models with vacancies. At bulk tricriticality, we find that the open edges are dominated by the vacancies when the surface coupling K(s) and the chemical potential D(s) of the vacancies assume the bulk values. When K(s) and/or D(s) is sufficiently enhanced, an edge phase transition takes place, beyond which spontaneous one-dimensional order occurs on the edges. Edge phase transitions can also be induced by a surface magnetic field H(s) . We numerically determine a number of edge critical exponents and derive phase diagrams in terms of K(s) , D(s) , and H(s) . In the low-temperature region, we observe first-order transitions when K(s) and D(s) are varied; the associated hysteresis loops of surface quantities are remarkably asymmetric. Some further insight into these edge transitions is provided by the exact equivalence of the tricritical q=1 Potts model and the Ising model. PMID- 15783382 TI - Spectral moments of correlated Wishart matrices. AB - We present an analytic method to determine the spectral properties of the covariance matrices constructed of correlated Wishart random matrices. The method gives, in the limit of large matrices, exact analytic relations between the spectral moments and the eigenvalue densities of the covariance matrices and their estimators. The results can be used in practice to extract the information about genuine correlations from the given experimental realization of random matrices. PMID- 15783383 TI - Emergence of large-scale vorticity during diffusion in a random potential under an alternating bias. AB - Conventional wisdom indicates that the presence of an alternating driving force will not change the long-term behavior of a Brownian particle moving in a random potential. Although this is true in one dimension, here we offer direct evidence that the inevitable local symmetry breaking present in a two-dimensional random potential leads to the emergence of a local ratchet effect that generates large scale vorticity patterns consisting of steady-state net diffusive currents. For small fields the spatial correlation function of the current follows a logarithmic distance dependence, while for large external fields both the vorticity and the correlations gradually disappear. We uncover the scaling laws characterizing this unique pattern formation process, and discuss their potential relevance to real systems. PMID- 15783381 TI - Nonuniversal coarsening and universal distributions in far-from-equilibrium systems. AB - Anomalous coarsening in far-from-equilibrium one-dimensional systems is investigated by applying simulation and analytic techniques to minimal hard-core particle (exclusion) models. They contain mechanisms of aggregated particle diffusion, with rates epsilon<<1 , particle deposition into cluster gaps, but suppressed for the smallest gaps, and breakup of clusters that are adjacent to large gaps. Cluster breakup rates vary with the cluster length x as k x(alpha) . The domain growth law x approximately (epsilont)(z) , with z=1/ (2+alpha) for alpha>0 , is explained by a simple scaling picture involving the time for two particles to coalesce and a new particle to be deposited. The density of double vacancies, at which deposition and cluster breakup are allowed, scales as 1/ [t (epsilont)(z) ] . Numerical simulations for several values of alpha and epsilon confirm these results. A fuller approach is presented which employs a mapping of cluster configurations to a column picture and an approximate factorization of the cluster configuration probability within the resulting master equation. The equation for a one-variable scaling function explains the above average cluster length scaling. The probability distributions of cluster lengths x scale as P (x) = [1/ (epsilont)(z) ] g (y) , with y identical with x/ (epsilont)(z) , which is confirmed by simulation. However, those distributions show a universal tail with the form g (y) approximately exp (- y(3/2) ) , which is explained by the connection of the vacancy dynamics with the problem of particle trapping in an infinite sea of traps. The high correlation of surviving particle displacement in the latter problem explains the failure of the independent cluster approximation to represent those rare events. PMID- 15783385 TI - Energy landscape and rigidity. AB - The effects of floppy modes in the thermodynamical properties of a system are studied. From thermodynamical arguments, we deduce that floppy modes are not at zero frequency and thus a modified Debye model is used to take into account this effect. The model predicts a deviation from the Debye law at low temperatures. Then, the connection between the topography of the energy landscape, the topology of the phase space, and the rigidity of a glass is explored. As a result, we relate the number of constraints and floppy modes to the statistics of the landscape. We apply these ideas to a simple model for which we provide an approximate expression for the number of energy basins as a function of the rigidity. This helps to understand certain features of the glass transition, like the jump in the specific heat or the reversible window observed in chalcogenide glasses. PMID- 15783384 TI - Generating function, path integral representation, and equivalence for stochastic exclusive particle systems. AB - We present the path integral representation of the generating function for classical exclusive particle systems. By introducing hard-core bosonic creation and annihilation operators and appropriate commutation relations, we construct the Fock space structure. Using the state vector, the generating function is defined and the master equation of the system is transformed into the equation for the generating function. Finally, the solution of the linear equation for the generating function is derived in the form of the path integral. Applying the formalism, the equivalence of reaction-diffusion processes of single species and two species is described. PMID- 15783386 TI - Brownian simulations and unidirectional flux in diffusion. AB - The prediction of ionic currents in protein channels of biological membranes is one of the central problems of computational molecular biophysics. Existing continuum descriptions of ionic permeation fail to capture the rich phenomenology of the permeation process, so it is therefore necessary to resort to particle simulations. Brownian dynamics (BD) simulations require the connection of a small discrete simulation volume to large baths that are maintained at fixed concentrations and voltages. The continuum baths are connected to the simulation through interfaces, located in the baths sufficiently far from the channel. Average boundary concentrations have to be maintained at their values in the baths by injecting and removing particles at the interfaces. The particles injected into the simulation volume represent a unidirectional diffusion flux, while the outgoing particles represent the unidirectional flux in the opposite direction. The classical diffusion equation defines net diffusion flux, but not unidirectional fluxes. The stochastic formulation of classical diffusion in terms of the Wiener process leads to a Wiener path integral, which can split the net flux into unidirectional fluxes. These unidirectional fluxes are infinite, though the net flux is finite and agrees with classical theory. We find that the infinite unidirectional flux is an artifact caused by replacing the Langevin dynamics with its Smoluchowski approximation, which is classical diffusion. The Smoluchowski approximation fails on time scales shorter than the relaxation time 1/gamma of the Langevin equation. We find that the probability of Brownian trajectories that cross an interface in one direction in unit time Deltat equals that of the probability of the corresponding Langevin trajectories if gammaDeltat=2 . That is, we find the unidirectional flux (source strength) needed to maintain average boundary concentrations in a manner consistent with the physics of Brownian particles. This unidirectional flux is proportional to the concentration and inversely proportional to sqrt[Deltat ] to leading order. We develop a BD simulation that maintains fixed average boundary concentrations in a manner consistent with the actual physics of the interface and without creating spurious boundary layers. PMID- 15783387 TI - Effect of random internal structure on combustion of binary powder mixtures. AB - Combustion of a mixture of two solid reactants is considered. A combined geometrical and physicochemical model of mixing of these reactants is proposed. The model takes into account random distribution of reactants and voids over the system. This allows description of incomplete burning of reactants. The model is used for studying the combustion wave propagation in a binary heterogeneous mixture. The obtained results are compared with those calculated from the perfect mixing model, which implies that the deficient (with respect to the stoichiometry) component burns completely. It is shown that micrononuniformity of mixing not only reduces the combustion wave temperature and velocity but also can lead to a shift of the maximal wave velocity away from the stoichiometric composition of the mixture. The results of this study allow us to suggest that one of the main reasons for such a shift, which was observed in a number of experimental works, is structural disorder of binary mixtures. PMID- 15783388 TI - Subgraphs and network motifs in geometric networks. AB - Many real-world networks describe systems in which interactions decay with the distance between nodes. Examples include systems constrained in real space such as transportation and communication networks, as well as systems constrained in abstract spaces such as multivariate biological or economic data sets and models of social networks. These networks often display network motifs: subgraphs that recur in the network much more often than in randomized networks. To understand the origin of the network motifs in these networks, it is important to study the subgraphs and network motifs that arise solely from geometric constraints. To address this, we analyze geometric network models, in which nodes are arranged on a lattice and edges are formed with a probability that decays with the distance between nodes. We present analytical solutions for the numbers of all three- and four-node subgraphs, in both directed and nondirected geometric networks. We also analyze geometric networks with arbitrary degree sequences and models with a bias for directed edges in one direction. Scaling rules for scaling of subgraph numbers with system size, lattice dimension, and interaction range are given. Several invariant measures are found, such as the ratio of feedback and feed forward loops, which do not depend on system size, dimension, or connectivity function. We find that network motifs in many real-world networks, including social networks and neuronal networks, are not captured solely by these geometric models. This is in line with recent evidence that biological network motifs were selected as basic circuit elements with defined information-processing functions. PMID- 15783389 TI - Critical dynamics of stochastic models with two conserved densities (model C' ). AB - We calculate the field-theoretic functions of the generalized dynamical model C(*') , where two conserved secondary densities are coupled to a nonconserved complex order parameter (OP), in two-loop order. A transformation to "orthogonalized" densities can be performed where only one secondary density with nontrivial static coupling to the OP exists while the second one remains Gaussian. The secondary densities remain dynamically coupled by the nondiagonal diffusion coefficent. General relations for the field-theoretic functions allow us to relate the asymptotic critical properties of model C(*') to the simpler model C(*) with only one conserved density coupled to the OP. The nonasymptotic properties, however, differ as can be seen from the flow of the dynamic parameters, which is presented for the case of a real OP with componets n=1,2,3 . PMID- 15783391 TI - Hysteresis multicycles in nanomagnet arrays. AB - We predict two physical effects in arrays of single-domain nanomagnets by performing simulations using a realistic model Hamiltonian and physical parameters. First, we find hysteretic multicycles for such nanomagnets. The simulation uses continuous spin dynamics through the Landau-Lifshitz-Gilbert (LLG) equation. In some regions of parameter space, the probability of finding a multicycle is as high as approximately 0.6 . We find that systems with larger and more anisotropic nanomagnets tend to display more multicycles. Our results also demonstrate the importance of disorder and frustration for multicycle behavior. Second, we show that there is a fundamental difference between the more realistic vector LLG equation and scalar models of hysteresis, such as Ising models. In the latter case spin and external field inversion symmetry is obeyed, but in the former it is destroyed by the dynamics, with important experimental implications. PMID- 15783392 TI - Directed percolation with long-range interactions: Modeling nonequilibrium wetting. AB - It is argued that some phase transitions observed in models of nonequilibrium wetting phenomena are related to contact processes with long-range interactions. This is investigated by introducing a model where the activation rate of a site at the edge of an inactive island of length l is 1+a l(-sigma) . Mean-field analysis and numerical simulations indicate that for sigma>1 the transition is continuous and belongs to the universality class of directed percolation, while for 01) and nonextensive (alpha<1) regimes. Results for finite size systems, taking into account all interactions between spins up to 24 sites apart, show that the conjecture is satisfied with a very good precision (less than 0.004%) for all temperature intervals. This TM procedure is further used to investigate several other thermodynamic and critical properties of this system, and it may also be extended to similar one-dimensional long-range systems. PMID- 15783398 TI - Exact cluster size distribution in the one-dimensional Ising model. AB - The exact solution for the cluster size distribution in the one-dimensional Ising model is obtained. In the thermodynamic limit the result is a simple analytical formula which gives the normalized number of clusters of different sizes. The analytical prediction is compared with Monte Carlo simulations and the energy dependence of the distribution is studied. PMID- 15783399 TI - Renormalization group study of a kinetically constrained model for strong glasses. AB - We derive a dynamic field theory for a kinetically constrained model, based on the Fredrickson-Andersen model, which we expect to describe the properties of an Arrhenius (strong) supercooled liquid at the coarse-grained level. We study this field theory using the renormalization group. For mesoscopic length and time scales, and for space dimension d>/=2 , the behavior of the model is governed by a zero-temperature dynamical critical point in the directed percolation universality class. We argue that in d=1 its behavior is that of compact directed percolation. We perform detailed numerical simulations of the corresponding Fredrickson-Andersen model on the lattice in various dimensions, and find reasonable quantitative agreement with the field theory predictions. PMID- 15783400 TI - Kinetic theory of random graphs: from paths to cycles. AB - The structural properties of evolving random graphs are investigated. Treating linking as a dynamic aggregation process, rate equations for the distribution of node to node distances (paths) and of cycles are formulated and solved analytically. At the gelation point, the typical length of paths and cycles, l , scales with the component size k as l approximately k(1/2) . Dynamic and finite size scaling laws for the behavior at and near the gelation point are obtained. Finite-size scaling laws are verified using numerical simulations. PMID- 15783401 TI - Dynamical real-space renormalization group calculations with a highly connected clustering scheme on disordered networks. AB - We have defined a type of clustering scheme preserving the connectivity of the nodes in a network, ignored by the conventional Migdal-Kadanoff bond moving process. In high dimensions, our clustering scheme performs better for correlation length and dynamical critical exponents than the conventional Migdal Kadanoff bond moving scheme. In two and three dimensions we find the dynamical critical exponents for the kinetic Ising model to be z=2.13 and z=2.09 , respectively, at the pure Ising fixed point. These values are in very good agreement with recent Monte Carlo results. We investigate the phase diagram and the critical behavior of randomly bond diluted lattices in d=2 and 3 in the light of this transformation. We also provide exact correlation exponent and dynamical critical exponent values on hierarchical lattices with power-law and Poissonian degree distributions. PMID- 15783402 TI - Self-similar variational perturbation theory for critical exponents. AB - We extend field theoretic variational perturbation theory by self-similar approximation theory, which greatly accelerates convergence. This is illustrated by recalculating the critical exponents of O (N) -symmetric phi(4) theory. From only three-loop perturbation expansions in 4-epsilon dimensions, we obtain analytic results for the exponents, which are close to those derived recently from ordinary field-theoretic variational perturbational theory to seventh order. In particular, the specific-heat exponent is found to be in good agreement with best-measured exponent alpha approximately -0.0127 of the specific-heat peak in superfluid helium, found in a satellite experiment. In addition, our analytic expressions reproduce also the exactly known large- N behavior of the exponents. PMID- 15783403 TI - Unidirectional drift of bistable front under asymmetrically oscillating zero-mean force. AB - The unidirectional drift of bistable fronts (BFs) that separate two stable uniform states of a bistable system under the action of an asymmetrically oscillating zero-mean force (driver) is considered within the "pseudolinear" (piecewise-linear) model of the system. The particular case of the symmetrical (symmetrically shaped) rate functions is studied. To perform a rigorous analytic treatment for arbitrary strengths of the driving force we assume that the applied ac force is quasistatically slow. Both cases of the initially static and the initially propagating BFs are examined; various types of the "unforced" dc motion are found. We show that the unforced transport of BF takes place in any case of the asymmetric driver, whether Maxwellian construction of the rate function was balanced or not. In particular, progressive (accelerated) dc drift of the initially static BFs occurred. In contrast, both progressive and regressive (decelerated) types of unforced dc drift of the initially propagating BFs take place. Moreover, reversal of the directed motion of the initially propagating BF occurred, if the deviation of Maxwellian construction from the strictly balanced situation was relatively small; by tuning the strength of the driving force the dc drift of BF exhibits the reversal. The symmetry properties of the biharmonic driver are discussed. The biharmonic ac force consisting of a superposition of the fundamental mode and its even (odd) superharmonics is an asymmetrically (symmetrically) oscillating one. The reversal type of the unforced dc drift occurred only in the case of the even superharmonic "mixing," when the superharmonic mode of the biharmonic driver was even. PMID- 15783404 TI - Effects of constant electric fields on the buoyant stability of reaction fronts. AB - The effects that applying constant electric fields have on the buoyant instability of reaction fronts propagating vertically in a Hele-Shaw cell are investigated for a range of electric field strengths and fluid parameters. The reaction produces a decrease in density across the front such that upwards propagating fronts are buoyantly unstable in the field-free situation. The reaction kinetics are modeled by cubic autocatalysis. A linear stability analysis reveals that a positive electric field increases the stability of a reaction front and can stabilize an otherwise unstable front. A negative field has the opposite effect, making the reaction front more unstable. Numerical simulations of the full nonlinear problem confirm these predictions and show the development of cellular fingers on unstable fronts. These simulations show that the electric field effects on the reaction within the front can alter the fluid density so as to give the possibility of destabilizing an otherwise stable downward propagating front. PMID- 15783405 TI - Lyapunov spreading of semiclassical wave packets for the Lorentz gas: theory and applications. AB - We consider the quantum-mechanical propagator for a particle moving in a d dimensional Lorentz gas, with fixed, hard-sphere scatterers. To evaluate this propagator in the semiclassical region, and for times less than the Ehrenfest time, we express its effect on an initial Gaussian wave packet in terms of quantities analogous to those used to describe the exponential separation of trajectories in the classical version of this system. This result relates the spread of the wave packet to the rate of separation of classical trajectories, characterized by positive Lyapunov exponents. We consider applications of these results, first to illustrate the behavior of the wave-packet autocorrelation functions for wave packets on periodic orbits. The autocorrelation function can be related to the fidelity, or Loschmidt echo, for the special case that the perturbation is a small change in the mass of the particle. An exact expression for the fidelity, appropriate for this perturbation, leads to an analytical result valid over very long time intervals, inversely proportional to the size of the mass perturbation. For such perturbations, we then calculate the long-time echo for semiclassical wave packets on periodic orbits. PMID- 15783406 TI - Random matrices with correlated elements: a model for disorder with interactions. AB - The complicated interactions in the presence of disorder lead to a correlated randomization of states. The Hamiltonian as a result behaves like a multiparametric random matrix with correlated elements. We show that the eigenvalue correlations of these matrices can be described by the single parametric Brownian ensembles. The analogy helps us to reveal many important features of the level statistics in interacting systems, e.g., a critical point behavior different from that of noninteracting systems, the possibility of extended states even in one dimension, and a universal formulation of level correlations. PMID- 15783407 TI - Crash test for the restricted three-body problem. AB - The restricted three-body problem serves to investigate the chaotic behavior of a small body under the gravitational influence of two heavy primary bodies. We analyze numerically the phase space mixing of bounded motion, escape, and crash in this simple model of (chaotic) celestial mechanics. The presented extensive numerical analysis reveals a high degree of complexity. We extend the recently presented findings for the Copenhagen case of equal main masses to the general case of different primary body masses. Collisions of the small body onto the primaries are comparatively frequent, and their probability displays a scale-free dependence on the size of the primaries as shown for the Copenhagen case. Interpreting the crash as leaking in phase space the results are related to both chaotic scattering and the theory of leaking Hamiltonian systems. PMID- 15783408 TI - Directed chaotic transport in Hamiltonian ratchets. AB - We present a comprehensive account of directed transport in one-dimensional Hamiltonian systems with spatial and temporal periodicity. They can be considered as Hamiltonian ratchets in the sense that ensembles of particles can show directed ballistic transport in the absence of an average force. We discuss general conditions for such directed transport like a mixed classical phase space. A sum rule is derived which connects the contributions of different phase space components to transport. We show that regular ratchet transport can be directed against an external potential gradient while chaotic ballistic transport is restricted to unbiased systems. For quantized Hamiltonian ratchets we study transport in terms of the evolution of wave packets and derive a semiclassical expression for the distribution of level velocities which encode the quantum transport in the Floquet band spectra. We discuss the role of dynamical tunneling between transporting islands and the chaotic sea and the breakdown of transport in quantum ratchets with broken spatial periodicity. PMID- 15783409 TI - Amplification of information transfer in excitable systems that reside in a steady state near a bifurcation point to complex oscillatory behavior. AB - We study the amplification of information transfer in excitable systems. We show that excitable systems residing in a steady state near a bifurcation point to complex oscillatory behavior incorporate several frequencies that can be exploited for a resonant amplification of information transfer. In particular, for excitable neurons that reside in a steady state near a bifurcation point to elliptic bursting oscillations, we show that in addition to the resonant frequency of damped oscillations around the stable focus, another frequency exists that resonantly enhances large amplitude bursts and thus amplifies the information transfer in the system. This additional frequency cannot be found by the local stability analysis and has never been used for amplifying the information transfer in a system. The results obtained for elliptic bursting oscillations can be generalized also to other complex oscillators, such as parabolic or square-wave bursters. Additionally, the biological importance of presented results in the field of neuroscience is outlined. PMID- 15783410 TI - Surrogate test to distinguish between chaotic and pseudoperiodic time series. AB - In this paper a different algorithm is proposed to produce surrogates for pseudoperiodic time series. By imposing a few constraints on the noise components of pseudoperiodic data sets, we devise an effective method to generate surrogates. Unlike other algorithms, this method properly copes with pseudoperiodic orbits contaminated with linear colored observational noise. We will demonstrate the ability of this algorithm to distinguish chaotic orbits from pseudoperiodic orbits through simulation data sets from the Rossler system. As an example of application of this algorithm, we will also employ it to investigate a human electrocardiogram record. PMID- 15783411 TI - Limits on the excitable behavior of a semiconductor laser with optical feedback. AB - Recently, it was proposed that semiconductor lasers with optical feedback present a complex behavior that can be described as noise driven excitable. In this work we investigate in which region of parameter space this description is adequate. We conclude that the region of the parameter space in which the system displays noise driven excitable behavior is a subset of the region in which presents low frequency fluctuations. PMID- 15783412 TI - Asynchronous random Boolean network model based on elementary cellular automata rule 126. AB - This paper considers a simple Boolean network with N nodes, each node's state at time t being determined by a certain number k of parent nodes, which is fixed for all nodes. The nodes, with randomly assigned neighborhoods, are updated based on various asynchronous schemes. We make use of a Boolean rule that is a generalization of rule 126 of elementary cellular automata. We provide formulas for the probability of finding a node in state 1 at a time t for the class of asynchronous random Boolean networks (ARBN) in which only one node is updated at every time step, and for the class of generalized ARBNs (GARBN) in which a random number of nodes can be updated at each time point. We use simulation methods to generate consecutive states of the network for both the real system and the models under the various schemes. The results match well. We study the dynamics of the models through sensitivity of the orbits to initial values, bifurcation diagrams, and fixed point analysis. We show, both theoretically and by example, that the ARBNs generate an ordered behavior regardless of the updating scheme used, whereas the GARBNs have behaviors that range from order to chaos depending on the type of random variable used to determine the number of nodes to be updated and the parameter combinations. PMID- 15783413 TI - Three-dimensional mixed-wet random pore-scale network modeling of two- and three phase flow in porous media. I. Model description. AB - We present a three-dimensional network model to simulate two- and three-phase capillary dominated processes at the pore level. The displacement mechanisms incorporated in the model are based on the physics of multiphase flow observed in micromodel experiments. All the important features of immiscible fluid flow at the pore scale, such as wetting layers, spreading layers of the intermediate-wet phase, hysteresis, and wettability alteration are implemented in the model. Wettability alteration allows any values for the advancing and receding oil water, gas-water, and gas-oil contact angles to be assigned. Multiple phases can be present in each pore or throat (element), in wetting and spreading layers, as well as occupying the center of the pore space. In all, some 30 different generic fluid configurations for two- and three-phase flow are analyzed. Double displacement and layer formation are implemented as well as direct two-phase displacement and layer collapse events. Every element has a circular, square, or triangular cross section. A random network that represents the pore space in Berea sandstone is used in this study. The model computes relative permeabilities, saturation paths, and capillary pressures for any displacement sequence. A methodology to track a given three-phase saturation path is presented that enables us to compare predicted and measured relative permeabilities on a point-by-point basis. A robust displacement-based clustering algorithm is also presented. PMID- 15783414 TI - Three-dimensional mixed-wet random pore-scale network modeling of two- and three phase flow in porous media. II. Results. AB - We use the model described in Piri and Blunt [Phys. Rev. E 71, 026301 (2005)] to predict two- and three-phase relative permeabilities of Berea sandstone using a random network to represent the pore space. We predict measured relative permeabilities for two-phase flow in a water-wet system. We then successfully predict the steady-state oil, water, and gas three-phase relative permeabilities measured by Oak (Proceedings of the SPE/DOE Seventh Symposium on Enhanced Oil Recovery, Tulsa, OK, 1990). We also study secondary and tertiary gas injection into media of different wettability and initial oil saturation and interpret the results in terms of pore-scale displacement processes. PMID- 15783415 TI - Two-phase flow in porous media: Crossover from capillary fingering to compact invasion for drainage. AB - It had been predicted that the capillary fingering observed at small capillary numbers should change or cross over to compact invasion at larger capillary numbers or longer times [D. Wilkinson, Phys. Rev. A 34, 1380 (1986)]. We present results from pore-level modeling in two dimensions for the average position (related to the position of the interface) of the injected fluid as well as the width of the interface between the injected, nonwetting fluid and the defending, wetting fluid. These results are entirely consistent with the predicted crossover from the fractal flow characterized by invasion percolation with trapping (IPWT) to compact/linear/stable flow, where the position of the injected fluid advances linearly with time and where the width of the interface is constant. Furthermore, our results for the characteristic time, at which the crossover occurs, agree with the predictions of Wilkinson. To focus on the effect of capillary number, we are considering only viscosity-matched flows where both fluids have the same viscosities. To our knowledge, these are the first pore-level modeling results that quantitatively test the general predictions of Wilkinson for this capillary crossover in the case of drainage. Our modeling results are used to provide closed form expressions predicting the dependence of average position and interfacial width upon capillary number and time, regardless of the size of the system. The size scaling inherent in the crossover combined with our results locating the upper and lower bounds of the crossover regime enable us to predict the location of the crossover for two-dimensional systems of different size. These predictions are compared with flow patterns from experiments in the literature. The agreement between our predictions and the experimental flow patterns indicates that the experiments exhibit the same IPWT to compact crossover observed in our modeling. PMID- 15783416 TI - Unstable diffusion and chemical dissociation of a single sonoluminescing bubble. AB - In a certain parameter region, a single sonoluminescencing bubble is unstable against diffusion of gases and their chemical dissociation. Experiments show that a surface unstable bubble emits a microbubble and recoils. After this it exhibits specific dynamical features whereby the ambient radius changes in a nonmonotonic way. A numerical analysis identifies the phenomenon as the result of the interplay between spatial translations and induced variations of driving pressure on one side and the chemical composition of gases in the bubble on the other side. The results confirm that dynamical chemical dissociation phenomena as well as acoustic properties play an important role in the understanding of single bubble sonoluminescence. PMID- 15783417 TI - Three-dimensional modes in a periodically driven elongated cavity. AB - Three-dimensional instability modes of the periodic flow in a rectangular cavity driven by the harmonic sliding oscillation of its floor are explored experimentally. Theory for a cavity with infinite span predicts two synchronous modes and a quasiperiodic traveling-wave mode as primary transitions from two dimensional to three-dimensional flow for different combinations of floor oscillation amplitude and frequency. Previously, only one of the two synchronous modes had been found experimentally. Here, we provide experimental details of both synchronous modes and a quasiperiodic mode. All three modes appear in the parameter regimes predicted by the theory; however, in the finite-span experiments, the traveling wave nature of the quasiperiodic mode is replaced by a nonpropagating mode with spatial features similar to those of the traveling mode. PMID- 15783418 TI - Stability of stratified flow with inhomogeneous shear. AB - The temporal evolution of perturbations in stratified flow with inhomogeneous shear is examined analytically by an extension of the nonmodal approach to flows with inhomogeneous shear. The solutions of the equations that govern the linear evolution and the weak nonlinear evolution of perturbations of the stream function for stratified flow with monotonic inhomogeneous shear are obtained. It is shown that stabilization of perturbations arises from nonmodal effects due to flow shear. Conditions at which these nonmodal effects may be strong enough to stabilize the Rayleigh-Taylor instability are presented. These analytical results are also compared to numerical simulations of the governing equations performed by Benilov, Naulin, and Rasmussen. PMID- 15783419 TI - Nonlinear dynamics of short traveling capillary-gravity waves. AB - We establish a Green-Nagdhi model equation for capillary-gravity waves in (2+1) dimensions. Through the derivation of an asymptotic equation governing short-wave dynamics, we show that this system possesses (1+1) traveling-wave solutions for almost all the values of the Bond number theta (the special case theta=1/3 is not studied). These waves become singular when their amplitude is larger than a threshold value, related to the velocity of the wave. The limit angle at the crest is then calculated. The stability of a wave train is also studied via a Benjamin-Feir modulational analysis. PMID- 15783420 TI - Signature of elasticity in the Faraday instability. AB - We investigate the onset of the Faraday instability in a vertically vibrated wormlike micelle solution. In this strongly viscoelastic fluid, the critical acceleration and wave number are shown to present oscillations as a function of driving frequency and fluid height. This effect, unseen in either in simple fluids or in previous experiments on polymeric fluids, is interpreted in terms of standing elastic waves between the disturbed surface and the container bottom. It is shown that the model of Kumar [Phys. Rev. E 65, 026305 (2002)] for a viscoelastic fluid accounts qualitatively for our experimental observations. Explanations for quantitative discrepancies are proposed, such as the influence of the nonlinear rheological behavior of this complex fluid. PMID- 15783421 TI - Finite-size scaling of two-point statistics and the turbulent energy cascade generators. AB - Within the framework of random multiplicative energy cascade models of fully developed turbulence, finite-size-scaling expressions for two-point correlators and cumulants are derived, taking into account the observationally unavoidable conversion from an ultrametric to an Euclidean two-point distance. The comparison with two-point statistics of the surrogate energy dissipation, extracted from various wind tunnel and atmospheric boundary layer records, allows an accurate deduction of multiscaling exponents and cumulants, even at moderate Reynolds numbers for which simple power-law fits are not feasible. The extracted exponents serve as input for parametric estimates of the probabilistic cascade generator. Various cascade generators are evaluated. PMID- 15783422 TI - Instability of sonoluminescing bubbles under a nonspherical symmetrical acoustic pressure perturbation. AB - The perturbation of nonspherical symmetrical acoustic pressure is added to the equation governing the spherical stability of sonoluminescing bubbles. The numerical calculations of the shape instability of sonoluminescing bubbles with the modified equation are conducted and the results are illustrated accordingly in the p(a) - R0 phase diagrams. The calculated results indicate that the stability region vanishes as the amplitude of the driving acoustic pressure p(a) arrives at the upper threshold ( approximately 1.6 atm) due to the perturbation of a small nonspherical symmetrical acoustic pressure (about a few Pa), which is in consistence with the experimental observations. PMID- 15783423 TI - Quasineutral plasma models. AB - The quasineutral plasma model proposed by Langmuir more than 75 years ago is still widely used today and is based on two approximations: charge neutrality and the Boltzmann relationship for electrons. However, the Boltzmann relationship is unnecessary and is not always justified. Moreover, because the Boltzmann relationship is fluid based, it compromises kinetic treatments and gives rise to troublesome singularities in the Bohm condition. To overcome these limitations, more general quasineutral models are developed. Two of the models are fluid based while the third is fully kinetic. The kinetic model and one of the fluid models lead directly to the Bohm condition, but without the singularities seen earlier. Fluid simulations are presented to test and compare the various approaches. PMID- 15783424 TI - Free-energy model for fluid helium at high density. AB - We present a semianalytical free-energy model aimed at characterizing the thermodynamic properties of dense fluid helium, from the low-density atomic phase to the high-density fully ionized regime. The model is based on a free-energy minimization method and includes various different contributions representative of the correlations between atomic and ionic species and electrons. This model allows the computation of the thermodynamic properties of dense helium over an extended range of density and temperature and leads to the computation of the phase diagram of dense fluid helium, with its various temperature and pressure ionization contours. One of the predictions of the model is that pressure ionization occurs abruptly at rho greater, > or = 10 g cm(-3) , i.e., P greater, > or = 20 Mbar , from atomic helium He to fully ionized helium He2+ , or at least to a strongly ionized state, without a He+ stage, except at high enough temperature for temperature ionization to become dominant. These predictions and this phase diagram provide a guide for future dynamical experiments or numerical first-principle calculations aimed at studying the properties of helium at very high density, in particular its metallization. Indeed, the characterization of the helium phase diagram bears important consequences for the thermodynamic, magnetic, and transport properties of cool and dense astrophysical objects, among which are the solar and the numerous recently discovered extrasolar giant planets. PMID- 15783425 TI - Effect of Coulomb scattering on low-pressure high-density electronegative discharges. AB - For electronegative plasmas with low gas pressure and high ion densities, we expect Coulomb collisions between positive and negative ions to dominate over collisions between ions and neutrals. We incorporated Nanbu's cumulative small angle collision method [K. Nanbu, Phys. Rev. E, 55, 4642 (1997)] into our one dimensional three-velocity-component particle-in-cell code PDP1 in order to study the effect of Coulomb collisions on low pressure high density electronegative discharges. Nanbu's method treats a succession of small-angle binary collisions as a single binary collision with a large scattering angle, which is far faster than treating each individual small-angle collision. We find that Coulomb collisions between positive and negative ions in low-pressure high-density electronegative discharges significantly modify the negative ion flux, density, and kinetic energy profiles. PMID- 15783426 TI - Quasimonoenergetic electrons from unphased injection into channel guided laser wakefield accelerators. AB - A high-quality electron beam can be extracted from a channel guided laser wakefield accelerator without confining the injected particles to a small region of phase. By careful choice of the injection energy, a regime can be found where uniformly phased particles are quickly bunched by the accelerator itself and subsequently accelerated to high energy. The process is particularly effective in a plasma channel because of a favorable phase shift that occurs in the focusing fields. Furthermore, particle-in-cell simulations show that the self-fields of the injected bunches actually tend to reduce the energy spread on the final beam. The final beam characteristics can be calculated using a computationally inexpensive Hamiltonian formulation when beam-loading effects are minimal. PMID- 15783427 TI - Effect of ambipolar fluxes on nanoparticle charging in low-pressure glow discharges. AB - The effect of ambipolar fluxes on nanoparticle charging in a typical low-pressure parallel-plate glow discharge is considered. It is shown that the equilibrium values of the nanoparticle charge in the plasma bulk and near-electrode areas are strongly affected by the ratio sigma(i)(ath) of the ambipolar flux and the ion thermal velocities. Under typical experimental conditions the above ratio is neither sigma(i)(ath)<<1 nor sigma(i)(ath)>>1 , which often renders the commonly used approximations of the purely thermal or "ion wind" ion charging currents inaccurate. By using the general approximation for the ambipolar drift-affected ion flux on the nanoparticle surface, it appears possible to obtain more accurate values of the nanoparticle charge that usually deviate within 10-25 % from the values obtained without a proper accounting for the ambipolar ion fluxes. The implications of the results obtained for glow discharge modeling and nanoparticle manipulation in low-pressure plasmas are discussed. PMID- 15783428 TI - Structural phase transitions and out-of-plane dust lattice instabilities in vertically confined plasma crystals. AB - The formation of plasma crystals confined in an external one-dimensional parabolic potential well is simulated for a normal experimental environment employing a computer code called BOX_TREE. Under appropriate conditions, crystals were found to form layered systems. The system's structural phase transitions, including transitions between crystals with differing numbers of layers and the same number of layers but different intralayer structures, were investigated and found to agree with previous theoretical and experimental research results. One- to two-layer transitions were examined in detail and shown to start at the point where the out-of-plane lattice instability appears. The resulting three layer system caused by this instability was observed at the center of the system. Finally, growth rates for this out-of-plane lattice instability were obtained using the BOX_TREE simulation with these results shown to agree with those obtained from analytical theory. PMID- 15783429 TI - Stopping of relativistic electrons in a partially degenerate electron fluid. AB - The stopping mechanisms of relativistic electron beams in superdense and partially degenerate electron fluid targets are investigated in the framework of the fast ignitor concept for inertial confinement fusion. In order to comply with specific demands in this area, we focus attention on the target partial degeneracy parameter theta= T(e) / T(f) , in terms of the thermal to Fermi temperature ratio. The target electron fluid is thus modeled very accurately with a random phase approximation dielectric function. The stopping results are shown to be very weakly theta dependent. However, a quantum target description is needed to recover their correct increasing trend with increasing projectile energy. The ranges and effective penetration depths in precompressed thermonuclear fuels are shown to be nearly a factor of 2 shorter than earlier classical estimates in the same conditions. The overall conclusions pertaining to the feasibility of fast ignition thus remain unchanged. PMID- 15783430 TI - Collisional cross sections and momentum distributions in astrophysical plasmas: dynamics and statistical mechanics link. AB - We show that in stellar core plasmas, the one-body momentum distribution function is strongly dependent, at least in the high velocity regime, on the microscopic dynamics of ion elastic collisions and therefore on the effective collisional cross sections if a random force field is present. We take into account two cross sections describing ion-dipole and ion-ion screened interactions. Furthermore, we introduce a third unusual cross section to link statistical distributions and a quantum effect originated by the energy-momentum uncertainty owing to many-body collisions. We also propose a possible physical interpretation in terms of a tidal-like force. We show that each collisional cross section gives rise to a slight peculiar correction on the Maxwellian momentum distribution function in a well defined velocity interval. We also find a possible link between microscopic dynamics of ions and statistical mechanics in interpreting our results in the framework of nonextensive statistical mechanics. PMID- 15783431 TI - Characterization of a vacuum-arc discharge in tin vapor using time-resolved plasma imaging and extreme ultraviolet spectrometry. AB - Discharge sources in tin vapor have recently been receiving increased attention as candidate extreme ultraviolet (EUV) light sources for application in semiconductor lithography, because of their favorable spectrum near 13.5 nm. In the ASML EUV laboratory, time-resolved pinhole imaging in the EUV and two dimensional imaging in visible light have been applied for qualitative characterization of the evolution of a vacuum-arc tin vapor discharge. An EUV spectrometer has been used to find the dominant ionization stages of tin as a function of time during the plasma evolution of the discharge. PMID- 15783432 TI - Vertical wave packets observed in a crystallized hexagonal monolayer complex plasma. AB - Propagation of vertical wave packets was observed experimentally in a crystallized hexagonal monolayer complex plasma. It was found that the phase velocity exceeded the group velocity by a factor 65 and was directed into the opposite direction as expected for an inverse optical-like dispersion relation. The wave packets propagated keeping their width constant. The explanation of this behavior is based on three-dimensional equations of motion and uses a long wavelength weak dispersion weak inhomogeneity approximation. While the wave dispersion causes the wave packet to spread, lattice inhomogeneity and neutral gas drag counteract spreading. A plasma diagnostic method was developed that is based on the ratio between vertical and dust-lattice wave speeds. This ratio is very sensitive to the lattice parameter kappa (ratio of the particle separation to the screening length) in a very useful range of kappa < or = 2 . It was found that only a two-dimensional lattice model can provide a quantitative description of the vertical waves, while a linear chain model gives only a qualitative agreement. PMID- 15783433 TI - Symmetric and asymmetric evaluation of the Tokamap in comparison with direct symplectic integration. AB - Comparing the Poincare plots of the Tokamap and the underlying Hamiltonian system reveals large differences. This stems from the particular choice of evaluation of the singular perturbations present in the system (a series of delta functions). A symmetric evaluation approach is proposed and shown to yield results that almost perfectly match the Hamiltonian system. PMID- 15783434 TI - Longitudinal impedance and shielding effectiveness of a resistive beam pipe for arbitrary energy and frequency. AB - The longitudinal coupling impedance of a cylindrical beam pipe for arbitrary relativistic gamma(0) and mode frequency is obtained analytically for finite wall conductivity and finite wall thickness. Closed form expressions for the electromagnetic fields excited by a beam perturbation are derived analytically. General expressions for the resistive-wall impedance in the presence of a metallic shield and for the rf shielding effectiveness of the beam pipe have been obtained and then compared with approximate expressions. The results are applied to the GSI synchrotron SIS, where the thickness of the vacuum chamber in the dipole magnets is much smaller than the skin depth at injection energy. PMID- 15783435 TI - Spontaneous one- and two-dimensional optical spatial solitons supported by photoisomerization nonlinearity in a bulk polymer. AB - We present theoretically another type of optical spatial soliton in steady state, which is supported by photoisomerization nonlinearity in a polymer. For the photoisomerization nonlinearity with saturation, this type of one- or two dimensional spatial soliton can be formed in a bulk polymer. The spontaneous soliton of this type can only be dark or gray. Not only a linearly polarized beam, but also a circularly polarized beam can be used to form the soliton, which is impossible for the photorefractive soliton. The effects of wavelength, polarization, and intensity of the beam on the formation of the soliton are discussed in detail. PMID- 15783436 TI - Enhancement of microcavity lifetimes using highly dispersive materials. AB - We show analytically and numerically that highly dispersive media can be used to drastically increase lifetimes of high- Q microresonators. In such a resonator, lifetime is limited either by undesired coupling to radiation, or by intrinsic absorption of the constituent materials. The presence of dispersion weakens coupling to the undesired radiation modes and also effectively reduces the material absorption. PMID- 15783437 TI - Acceleration of electrons from rest to GeV energies by ultrashort transverse magnetic laser pulses in free space. AB - In this paper we describe a laser acceleration scheme where an electron is accelerated from rest to GeV energies by the longitudinal electric field of an ultrashort transverse magnetic ( TM01 ) optical pulse. The on-axis longitudinal electric field of the pulse is obtained from the free-space divergence equation beyond the so-called slowly-varying-envelope approximation. The instantaneous electron dynamics is studied; numerical simulations predict net energy gains in the GeV range for laser intensities reaching 10(22) W/ cm(2) . PMID- 15783438 TI - Hybrid superprism with low insertion losses and suppressed cross-talk. AB - We demonstrate with the two-dimensional finite-difference time-domain method that an adiabatic transition in a superprism with an interface along the [1 2 ] direction enhances the transmission through the superprism to more than 90% (-0.5 dB) over the wavelength range 1.47-1.68 mum , including the telecommunication C and L bands. We also show that diffraction governed by a quasinegative index of refraction inside the superprism can be used to obtain nearly transform-limited beam widths at the output of the superprism. The reduction of the beam width at the output suppresses cross-talk and greatly enhances the achievable frequency resolution of the superprism. PMID- 15783439 TI - Diffractionless flow of light in two- and three-dimensional photonic band gap heterostructures: Theory, design rules, and simulations. AB - We demonstrate on-chip, single-mode, waveguiding of light in air for a variety of 2D-3D photonic band gap (PBG) heterostructures. These include square spiral, woodpile, slanted pore, and inverse opal three-dimensional (3D) photonic crystals intercalated with a 2D (planar) photonic crystal microchip. Design rules are established to yield maximal single-mode waveguiding bandwidths of up to roughly 180 nanometers centered at a wavelength of 1.5 microns . This can be achieved with 3D PBG materials with gaps as small as 15% of the PBG center frequency. Finite-difference time-domain (FDTD) simulations of light flow in optical mirocircuits within such heterostructures reveal tolerance to layer misalignment and other fabrication-related structural disorder. We provide an interpretation of the universal mechanism for diffractionless light propagation in 2D-3D photonic crystal heterostructures. We demonstrate that planar, on-chip, optical microcircuitry similar to that of two-dimensional (2D) photonic crystals is almost universally achievable within the engineered electromagnetic vacuum of 3D PBG materials. PMID- 15783440 TI - Analytical tools for solitons and periodic waves corresponding to phonons on Lennard-Jones lattices in helical proteins. AB - We study the propagation of solitons along the hydrogen bonds of an alpha helix. Modeling the hydrogen and peptide bonds with Lennard-Jones potentials, we show that the solitons can appear spontaneously and have long lifetimes. Remarkably, even if no explicit solution is known for the Lennard-Jones potential, the solitons can be characterized analytically with a good quantitative agreement using formulas for a Toda potential with parameters fitted to the Lennard-Jones potential. We also discuss and show the robustness of the family of periodic solutions called cnoidal waves, corresponding to phonons. The soliton phenomena described in the simulations of alpha helices may help to explain recent x-ray experiments on long alpha helices in Rhodopsin where a long lifetime of the vibrational modes has been observed. PMID- 15783441 TI - Nonlinear magnetoacoustic waves in rare-earth orthoferrites. AB - The dynamics of magnetoacoustic waves in a rare-earth orthoferrite YFeO3 crystal is experimentally investigated. The waves are excited by a high-power laser pulse. The analysis of results is carried out taking into account the nonlinear interaction of oscillation modes. It allows us to interpret the results as energy exchange between modes in the time scale, a phenomenon known in nonlinear optics. Based on the proposed model an estimation is obtained for effective nonlinear magnetoacoustic modules of fourth order for yttrium orthoferrite approximately 2.5x 10(17) -2.5x 10(19) J/ m(3) . PMID- 15783442 TI - Nonlinear compression of solitary waves in asymmetric twin-core fibers. AB - We demonstrate a different pulse compression technique based on exact solutions to the nonlinear Schrodinger-type equation interacting with a source, variable dispersion, variable Kerr nonlinearity, and variable gain or loss. We show that this model is appropriate for the pulse propagation in asymmetric twin-core fibers. The chirped pulses are compressed due to the nonlinearity as well as dispersion management as also due to the space dependence of the gain coefficient. We also obtain singular solitary wave solutions, pertaining to extreme increase of the amplitude due to self-focusing. PMID- 15783443 TI - Switching intense laser pulses guided by Kerr-effect-modified modes of a hollow core photonic-crystal fiber. AB - A Kerr-nonlinearity-induced profile of the refractive index in the hollow core of a photonic-crystal fiber (PCF) changes the spectrum of propagation constants of air-guided modes, effectively shifting the passbands in fiber transmission, controlled by the photonic band gaps (PBGs) of the cladding. This effect is shown to allow the creation of fiber switches for high-intensity laser pulses. The Kerr nonlinearity control of air-guided modes in PCFs and the performance of a PCF switch are quantified by solving the propagation equation for the slowly varying envelope of a laser pulse guided in Kerr-effect-modified PCF modes. The spatial dynamics of the light field in a PBG waveguide switch is analyzed with the use of the slowly varying envelope approximation, demonstrating high contrasts of optical switching with PBG waveguides and hollow PCFs. PMID- 15783444 TI - Large enhancement of third-harmonic generation induced by coupled gap solitons in chi(3) nonlinear photonic crystals. AB - It is shown that, in a chi((3)) nonlinear photonic crystal, the third-harmonic conversion efficiency can be enhanced by about three orders of magnitude via the formation of self-organized localized states inside the gaps as compared with that in a bulk medium of the same length with a perfect phase-matching condition. These localized states contain both the fundamental and third-harmonic frequencies located in different gaps. Their existence is a result of both the Kerr effect and the large energy transfer between fundamental and third-harmonic waves. PMID- 15783445 TI - Resonant transmission near nonrobust periodic slab modes. AB - We present a precise theoretical explanation and prediction of certain resonant peaks and dips in the electromagnetic transmission coefficient of periodically structured slabs in the presence of nonrobust guided slab modes. We also derive the leading asymptotic behavior of the related phenomenon of resonant enhancement near the guided mode. The theory applies to structures in which losses are negligible and to very general geometries of the unit cell. It is based on boundary-integral representations of the electromagnetic fields. These depend on the frequency and on the Bloch wave vector and provide a complex-analytic connection in these parameters between generalized scattering states and guided slab modes. The perturbation of three coincident zeros-those of the dispersion relation for slab modes, the reflection constant, and the transmission constant is central to calculating transmission anomalies both for lossless dielectric materials and for perfect metals. PMID- 15783446 TI - Localized modes in random arrays of cylinders. AB - Anderson localization of classical waves in random arrays of dielectric cylinders is numerically investigated as a function of the distribution of their diameters. We show that using polydispersed resonant scatterers increases the localization length, while using identical resonant scatterers fosters Anderson localization. We discuss this collective process and link it to the effect of proximity resonances that has been studied in the case of a small number of resonant scatterers. PMID- 15783447 TI - Complexes of stationary domain walls in the resonantly forced Ginsburg-Landau equation. AB - The parametrically driven Ginsburg-Landau equation has well-known stationary solutions-the so-called Bloch and Ne el, or Ising, walls. In this paper, we construct an explicit stationary solution describing a bound state of two walls. We also demonstrate that stationary complexes of more than two walls do not exist. PMID- 15783448 TI - Excitation of vortices using linear and nonlinear magnetostatic waves. AB - It is shown that stationary vortex structures can be excited in a ferrite film, in the important centimeter and millimeter wavelength ranges. It is shown that both linear and nonlinear structures can be excited using a three-beam interaction created with circular antennas. These give rise to a special phase distribution created by linear and nonlinear mixing. An interesting set of three clockwise rotating vortices joined by one counter-rotating one presents itself in the linear regime: a scenario that is only qualitatively changed by the onset of nonlinearity. It is pointed out that control of the vortex structure, through parametric coupling, based upon a microwave resonator, is possible and that there are many interesting possibilities for applications. PMID- 15783449 TI - Two-dimensional solitons with hidden and explicit vorticity in bimodal cubic quintic media. AB - We demonstrate that two-dimensional two-component bright solitons of an annular shape, carrying vorticities (m,+/-m) in the components, may be stable in media with the cubic-quintic nonlinearity, including the hidden-vorticity (HV) solitons of the type (m,-m) , whose net vorticity is zero. Stability regions for the vortices of both (m,+/-m) types are identified for m=1 , 2, and 3, by dint of the calculation of stability eigenvalues, and in direct simulations. In addition to the well-known symmetry-breaking (external) instability, which splits the ring soliton into a set of fragments flying away in tangential directions, we report two new scenarios of the development of weak instabilities specific to the HV solitons. One features charge flipping, with the two components exchanging angular momentum and periodically reversing the sign of their spins. The composite soliton does not directly split in this case; therefore, we identify such instability as an intrinsic one. Eventually, the soliton splits, as weak radiation loss drives it across the border of the ordinary strong (external) instability. Another scenario proceeds through separation of the vortex cores in the two components, each individual core moving toward the outer edge of the annular soliton. After expulsion of the cores, there remains a zero-vorticity breather with persistent internal vibrations. PMID- 15783450 TI - Finite-difference-based lattice Boltzmann model for dense binary mixtures. AB - We propose a finite-difference-based lattice Boltzmann model for dense binary mixtures based on the Enskog theory. The model is applicable to a mixture composed of two dense fluids with different shear viscosities. The macroscopic hydrodynamic and diffusion equations are derived from the model through the Chapmann-Enskog procedure. The model is also validated numerically. PMID- 15783451 TI - Spatially adaptive grand canonical ensemble Monte Carlo simulations. AB - A spatially adaptive Monte Carlo method is introduced directly from the underlying microscopic mechanisms, which satisfies detailed balance, gives the correct noise, and describes accurately dynamic and equilibrium states for adsorption-desorption (grand canonical ensemble) processes. It enables simulations of large scales while capturing sharp gradients with molecular resolution at significantly reduced computational cost. A posteriori estimates, in the sense used in finite-elements methods, are developed for assessing errors (information loss) in coarse-graining and guiding mesh generation. PMID- 15783452 TI - Test of a numerical approach to the quantization of billiards. AB - A method for computing large numbers of eigenvalues of self-adjoint elliptic operators [J. Comput. Phys. 184, 321 (2003)] is tested in numerical studies of the spectral properties of quantum billiards. To this extent, we study a time reversal invariant quantum billiard of threefold symmetry, that undergoes a transformation in its symmetry properties from C(3v) to C3 . Thereby a transition from Gaussian orthogonal to Gaussian unitary ensemble statistics is observed, verifying earlier experimental indications and theoretical predictions. At the same time our numerical ansatz is shown to be applicable to arbitrary billiard shapes. PMID- 15783453 TI - Parallel dynamics and computational complexity of network growth models. AB - The parallel computational complexity or depth of growing network models is investigated. The networks considered are generated by preferential attachment rules where the probability of attaching a new node to an existing node is given by a power alpha of the connectivity of the existing node. Algorithms for generating growing networks very quickly in parallel are described and studied. The sublinear and superlinear cases require distinct algorithms. As a result, there is a discontinuous transition in the parallel complexity of sampling these networks corresponding to the discontinuous structural transition at alpha=1 , where the networks become scale-free. For alpha>1 , networks can be generated in constant time while for 0infinity asymptotic decay of the total correlation function h (r) for a fluid composed of particles interacting via a (point) Yukawa pair potential. Such a potential provides a simple model for dusty plasmas. The asymptotic decay is determined by the poles of the liquid structure factor in the complex plane. We use the hypernetted-chain closure to the Ornstein-Zernike equation to determine the line in the phase diagram, well removed from the freezing transition line, where crossover occurs in the ultimate decay of h (r) , from monotonic to damped oscillatory. We show that (i) crossover takes place via the same mechanism (coalescence of imaginary poles) as in the classical one component plasma and in other models of Coulomb fluids and (ii) leading-order pole contributions provide an accurate description of h (r) at intermediate distances r as well as at long range. PMID- 15783461 TI - Band-gap engineering in two-dimensional semiconductor-dielectric photonic crystals. AB - This paper reports the multiple band gaps in the two-dimensional semiconductor dielectric photonic crystals of several compositions: semiconductor cylinders in the dielectric background. We consider both square lattice and hexagonal lattice arrangements and compute extensive band structures using a plane-wave method within the framework of an efficient standard eigenvalue problem for both E and H polarizations. The whole range of filling fraction has been explored to claim the existence of the lowest (the so-called acoustic) band gap and multiple higher energy band gaps within the first 30 to 40 bands for various compositions. Such semiconductor-dielectric photonic crystals which are shown to possess huge lowest band gaps below a threshold frequency (the plasma frequency omega(p) ) have an advantage over the dielectric photonic crystals in the emerging technology based on the photonic crystals. PMID- 15783462 TI - Open circular billiards and the Riemann hypothesis. AB - A comparison of escape rates from one and from two holes in an experimental container (e.g., a laser trap) can be used to obtain information about the dynamics inside the container. If this dynamics is simple enough one can hope to obtain exact formulas. Here we obtain exact formulas for escape from a circular billiard with one and with two holes. The corresponding quantities are expressed as sums over zeros of the Riemann zeta function. Thus we demonstrate a direct connection between recent experiments and a major unsolved problem in mathematics, the Riemann hypothesis. PMID- 15783463 TI - Exact ground state and finite-size scaling in a supersymmetric lattice model. AB - We study a model of strongly correlated fermions in one dimension with extended N = 2 supersymmetry. The model is related to the spin S = 1/2 XXZ Heisenberg chain at anisotropy Delta = -1/2 with a real magnetic field on the boundary. We exploit the combinatorial properties of the ground state to determine its exact wave function on finite lattices with up to 30 sites. We compute several correlation functions of the fermionic and spin fields. We discuss the continuum limit by constructing lattice observables with well defined finite-size scaling behavior. For the fermionic model with periodic boundary conditions we give the emptiness formation probability in closed form. PMID- 15783464 TI - Oscillations of a bose-einstein condensate rotating in a harmonic plus quartic trap. AB - We study the normal modes of a two-dimensional rotating Bose-Einstein condensate confined in a quadratic plus quartic trap. Hydrodynamic theory and sum rules are used to derive analytical predictions for the collective frequencies in the limit of high angular velocities Omega where the vortex lattice produced by the rotation exhibits an annular structure. We predict a class of excitations with frequency sqrt[6]Omega in the rotating frame, irrespective of the mode multipolarity m, as well as a class of low energy modes with frequency proportional to |m|/Omega. The predictions are in good agreement with results of numerical simulations based on the 2D Gross-Pitaevskii equation. The same analysis is also carried out at even higher angular velocities, where the system enters the giant vortex regime. PMID- 15783465 TI - Generation of entangled states of qudits using twin photons. AB - We report an experiment to generate entangled states of D-dimensional quantum systems, qudits, by using transverse spatial correlations of two parametric down converted photons. Apertures with D slits in the arms of the twin photons define the qudit space. By manipulating the pump beam correctly, the twin photons will pass only by symmetrically opposite slits, generating entangled states between these different paths. Experimental results for qudits with D = 4 and 8 are shown. We demonstrate that the generated states are entangled states. PMID- 15783466 TI - Geometric quantum computation and multiqubit entanglement with superconducting qubits inside a cavity. AB - We analyze a new scheme for quantum information processing, with superconducting charge qubits coupled through a cavity mode, in which quantum manipulations are insensitive to the state of the cavity. We illustrate how to physically implement universal quantum computation as well as multiqubit entanglement based on unconventional geometric phase shifts in this scalable solid-state system. Some quantum error-correcting codes can also be easily constructed using the same technique. In view of the gate dependence on just global geometric features and the insensitivity to the state of cavity modes, the proposed quantum operations may result in high-fidelity quantum information processing. PMID- 15783467 TI - Integration of Langevin equations with multiplicative noise and the viability of field theories for absorbing phase transitions. AB - Efficient and accurate integration of stochastic (partial) differential equations with multiplicative noise can be obtained through a split-step scheme, which separates the integration of the deterministic part from that of the stochastic part, the latter being performed by sampling exactly the solution of the associated Fokker-Planck equation. We demonstrate the computational power of this method by applying it to the most absorbing phase transitions for which Langevin equations have been proposed. This provides precise estimates of the associated scaling exponents, clarifying the classification of these nonequilibrium problems, and confirms or refutes some existing theories. PMID- 15783468 TI - Breakdown of an electric-field driven system: a mapping to a quantum walk. AB - Quantum transport properties of electron systems driven by strong electric fields are studied by mapping the Landau-Zener transition dynamics to a quantum walk on a semi-infinite one-dimensional lattice with a reflecting boundary, where the sites correspond to energy levels and the boundary the ground state. Quantum interference induces a distribution localized around the ground state, and a delocalization transition occurs when the electric field is increased, which describes the dielectric breakdown in the original electron system. PMID- 15783469 TI - 511 keV photons from color superconducting dark matter. AB - We discuss the possibility that the recent detection of 511 keV gamma rays from the galactic bulge, as observed by the International Gamma-Ray Astrophysics Laboratory, can be naturally explained by the supermassive very dense droplets (strangelets) of dark matter. These droplets are assumed to be made of ordinary light quarks (or antiquarks) condensed in a nonhadronic color superconducting phase. The droplets can carry electrons (or positrons) in the bulk or/and on the surface. The e(+)e(-) annihilation events take place due to the collisions of electrons from the visible matter with positrons from dark matter droplets which may result in the bright 511 keV gamma-ray line from the bulge of the Galaxy. PMID- 15783470 TI - Consistent discretization and loop quantum geometry. AB - We apply the "consistent discretization" approach to general relativity leaving the spatial slices continuous. The resulting theory is free of the diffeomorphism and Hamiltonian constraints, but one can impose the diffeomorphism constraint to reduce its space of solutions and the constraint is preserved exactly under the discrete evolution. One ends up with a theory that has as physical space what is usually considered the kinematical space of loop quantum geometry, given by diffeomorphism invariant spin networks endowed with appropriate rigorously defined diffeomorphism invariant measures and inner products. The dynamics can be implemented as a unitary transformation and the problem of time explicitly solved or at least reduced to a numerical problem. We exhibit the technique explicitly in (2+1)-dimensional gravity. PMID- 15783474 TI - Search for DeltaS = 2 nonleptonic hyperon decays. AB - A sensitive search for the rare decays Omega(-)--> Lambdapi(-) and Xi(0)--> ppi( ) has been performed using data from the 1997 run of the HyperCP (Fermilab E871) experiment. Limits on other such processes do not exclude the possibility of observable rates for |DeltaS| = 2 nonleptonic hyperon decays, provided the decays occur through parity-odd operators. We obtain the branching-fraction limits B(Omega(-)-->Lambdapi(-)) < 2.9 x 10(-6) and B(Xi(0)--> ppi(-)) < 8.2 x 10(-6), both at 90% confidence level. PMID- 15783475 TI - Measurement of the Michel parameter rho in muon decay. AB - The TWIST Collaboration has measured the Michel parameter rho in normal muon decay, mu(+)--> e(+)nu(e)nu (mu). In the standard model, rho = 3/4. Deviations from this value imply mixing of left- and right-handed muon and electron couplings. We find rho=0.750 80+/-0.000 32(stat) +/- 0.000 97(syst) +/- 0.000 23, where the last uncertainty represents the dependence of rho on the Michel parameter eta. This result sets new limits on the W(L)-W(R) mixing angle in left right symmetric models. PMID- 15783477 TI - Dissolving N = 4 loop amplitudes into QCD tree amplitudes. AB - We use the infrared consistency of one-loop amplitudes in N = 4 Yang-Mills theory to derive a compact analytic formula for a tree-level next-to-next-to-maximal helicity-violating gluon scattering amplitude in QCD, the first such formula known. We argue that the infrared conditions, coupled with recent advances in calculating one-loop box coefficients, can give a new tool for computing tree level amplitudes in general. Our calculation suggests that many amplitudes have a structure which is even simpler than that revealed so far by current twistor space constructions. PMID- 15783478 TI - Magnetic moment of the delta(1232) resonance in chiral effective-field theory. AB - We perform a relativistic chiral effective-field theory calculation of the radiative pion photoproduction (gammap--> pi(0)pgamma(')) in the Delta-resonance region, to next-to-leading order in the "delta expansion." This work is aimed at a model-independent extraction of the Delta(+) magnetic moment from new precise measurements of this reaction. It also predicts the chiral behavior of Delta's magnetic moment, which can be used to extrapolate the recent lattice QCD results to the physical point. PMID- 15783479 TI - Globally polarized quark-gluon plasma in noncentral A + A collisions. AB - Produced partons have a large local relative orbital angular momentum along the direction opposite to the reaction plane in the early stage of noncentral heavy ion collisions. Parton scattering is shown to polarize quarks along the same direction due to spin-orbital coupling. Such global quark polarization will lead to many observable consequences, such as left-right asymmetry of hadron spectra and global transverse polarization of thermal photons, dileptons, and hadrons. Hadrons from the decay of polarized resonances will have an azimuthal asymmetry similar to the elliptic flow. Global hyperon polarization is studied within different hadronization scenarios and can be easily tested. PMID- 15783480 TI - Quantum opacity, the RHIC Hanbury Brown-Twiss puzzle, and the chiral phase transition. AB - We present a relativistic quantum-mechanical treatment of opacity and refractive effects that allows reproduction of observables measured in two-pion Hanbury Brown-Twiss (HBT) interferometry and pion spectra at RHIC. The inferred emission duration is substantial. The results are consistent with the emission of pions from a system that has a restored chiral symmetry. PMID- 15783481 TI - Hard-loop dynamics of non-Abelian plasma instabilities. AB - Non-Abelian plasma instabilities may be responsible for the fast apparent quark gluon thermalization in relativistic heavy-ion collisions if their exponential growth is not hindered by nonlinearities. We study numerically the real-time evolution of instabilities in an anisotropic non-Abelian plasma with an SU(2) gauge group in the hard-loop approximation. We find exponential growth of non Abelian plasma instabilities both in the linear and in the strongly nonlinear regimes, with only a brief phase of subexponential behavior in between. PMID- 15783482 TI - Nuclear masses set bounds on quantum chaos. AB - It has been suggested that chaotic motion inside the nucleus may significantly limit the accuracy with which nuclear masses can be calculated. Using a power spectrum analysis we show that the inclusion of additional physical contributions in mass calculations, through many-body interactions or local information, removes the chaotic signal in the discrepancies between calculated and measured masses. Furthermore, a systematic application of global mass formulas and of a set of relationships among neighboring nuclei to more than 2000 nuclear masses allows one to set an unambiguous upper bound for the average errors in calculated masses, which turn out to be almost an order of magnitude smaller than estimated chaotic components. PMID- 15783483 TI - Fluctuations in the level density of a fermi gas. AB - We present a theory that accurately describes the counting of excited states of a noninteracting fermionic gas. At high excitation energies the results reproduce Bethe's theory. At low energies oscillatory corrections to the many-body density of states, related to shell effects, are obtained. The fluctuations depend nontrivially on energy and particle number. Universality and connections with Poisson statistics and random matrix theory are established for regular and chaotic single-particle motion. PMID- 15783484 TI - Systematics of quadrupolar correlation energies. AB - We calculate correlation energies associated with the quadrupole shape degrees of freedom with a view to improving the self-consistent mean-field theory of nuclear binding energies. Systematic results are presented for 605 even-even nuclei from mass number 16 to the heaviest whose mass has been measured, using the Skyrme SLy4 interaction and the generator coordinate method. Correlation energies range from 0.5 to 6.0 MeV, and their inclusion improves two qualitative deficiencies of the mean-field theory, namely, the exaggerated shell effect at neutron magic numbers and the failure of mean-field theory to describe mutually enhanced magicity. For the mass table as a whole, the quadrupolar correlations improve binding energies, separation energies, and separation energy differences by 20%30%. PMID- 15783485 TI - Precise determination of 6Li cold collision parameters by radio-frequency spectroscopy on weakly bound molecules. AB - We employ radio-frequency spectroscopy on weakly bound (6)Li(2) molecules to precisely determine the molecular binding energies and the energy splittings between molecular states for different magnetic fields. These measurements allow us to extract the interaction parameters of ultracold (6)Li atoms based on a multichannel quantum scattering model. We determine the singlet and triplet scattering lengths to be a(s) = 45.167(8)a(0) and a(t) = -2140(18)a(0) (1a(0) = 0.052 917 7 nm), and the positions of the broad Feshbach resonances in the energetically lowest three s-wave scattering channels to be 83.41(15), 69.04(5), and 81.12(10) mT. PMID- 15783471 TI - Search for the decay B+ --> K+nunu. AB - We search for the rare flavor-changing neutral-current decay B(+)--> K(+)nunu in a data sample of 82 fb(-1) collected with the BABAR detector at the PEP-II B factory. Signal events are selected by examining the properties of the system recoiling against either a reconstructed hadronic or semileptonic charged-B decay. Using these two independent samples we obtain a combined limit of B(B(+)- >K(+)nunu ) < 5.2 x 10(-5) at the 90% confidence level. In addition, by selecting for pions rather than kaons, we obtain a limit of B(B+-->pi(+)nunu) < 1.0 x 10( 4) using only the hadronic B reconstruction method. PMID- 15783486 TI - High-repetition-rate grazing-incidence pumped x-ray laser operating at 18.9 nm. AB - We have demonstrated a 10 Hz Ni-like Mo x-ray laser operating at 18.9 nm with 150 mJ total pump energy by employing a novel pumping scheme. The grazing-incidence scheme is described, where a picosecond pulse is incident at a grazing angle to a Mo plasma column produced by a slab target irradiated by a 200 ps laser pulse. This scheme uses refraction of the short pulse at a predetermined electron density to increase absorption to pump a specific gain region. The higher coupling efficiency inherent to this scheme allows a reduction in the pump energy where 70 mJ long pulse energy and 80 mJ short pulse energy are sufficient to produce lasing at a 10 Hz repetition rate. Under these conditions and by optimizing the delay between the pulses, we achieve strong amplification and close to saturation for 4 mm long targets. PMID- 15783487 TI - Experimental investigation of local properties and statistics of optical vortices in random wave fields. AB - We present the first direct experimental evidence of the local properties of optical vortices in a random laser speckle field. We have observed the Berry anisotropy ellipse describing the anisotropic squeezing of phase lines close to vortex cores and quantitatively verified the Dennis angular momentum rule for its phase. Some statistics associated with vortices, such as density, anisotropy ellipse eccentricity, and its relation to zero crossings of real and imaginary parts of the random field, are also investigated by experiments. PMID- 15783488 TI - Coherent focusing of high harmonics: a new way towards the extreme intensities. AB - We demonstrate analytically and numerically that focusing of high harmonics produced by the reflection of a few-femtosecond laser pulse from a concave plasma surface opens a new way to unprecedentally high intensities. The key features allowing the boosting of the focal intensity are the harmonics coherency and the small exponent of the power-law decay of the harmonics spectrum. Using similarity theory and direct particle-in-cell simulations, we find that the intensity at the focus scales as I(CHF) alpha a(3)(0)I(0), where a(0) and I(0) alpha a(2)(0) are the dimensionless relativistic amplitude and the intensity of the incident laser pulse. The scaling suggests that due to the coherent harmonic focusing (CHF), the Schwinger intensity limit can be reached using lasers with I(0) approximately 10(22) W/cm(2). The pulse duration at the focus scales as tau(CHF) alpha 1/a(2)(0) and reaches the subattosecond range. PMID- 15783490 TI - Nonlinear dynamics of transport barrier relaxations in tokamak edge plasmas. AB - A new mechanism for intermittent relaxations of transport barriers is found by using three dimensional fluid turbulence simulations. This mechanism is generic since it only requires a stationary E x B shear flow. It is found here that if the flow shear increases faster than linearly with heating power, the relaxation frequency decreases with power. An analytical study reveals that this nonlinear dynamics is governed by a time delay for effective velocity shear stabilization. PMID- 15783491 TI - Inductive current density perturbations to probe electron internal transport barriers in tokamaks. AB - Improved electron energy confinement in tokamak plasmas, related to internal transport barriers, has been linked to nonmonotonic current density profiles. This is difficult to prove experimentally since usually the current profiles evolve continuously and current injection generally requires significant input power. New experiments are presented, in which the inductive current is used to generate positive and negative current density perturbations in the plasma center, with negligible input power. These results demonstrate unambiguously for the first time that the electron confinement can be modified significantly solely by perturbing the current density profile. PMID- 15783489 TI - Design and fabrication of two-dimensional photonic crystals with predetermined nonlinear optical properties. AB - By probing the resonances between a photonic band and an external laser field and their nonlinear changes in angle-resolved reflectivity, we show experimental evidence that the nonlinear optical changes in a two-dimensional photonic crystal waveguide with a Kerr nonlinearity are critically dependent on the dispersion nature and the group velocity of the photonic bands. The results agree well with the behavior predicted from band structures, indicating that the design of nonlinear optical properties of material systems is realistically possible by band dispersion and group velocity engineering. PMID- 15783492 TI - Influence of the dynamic ergodic divertor on the density limit in TEXTOR. AB - A significant influence of the dynamic ergodic divertor (DED) on the density limit in TEXTOR has been found. In Ohmic discharges, where without DED detachment normally arises at the density limit, a MARFE (multifaceted asymmetric radiation from the edge) develops when the DED is operated in a static regime. The threshold of the MARFE onset in the neutral beam heated plasmas is increased by applying 1 kHz ac DED at the high-field side. The theoretical predictions based on the parallel energy balance taking poloidal asymmetries into account agree well with the experimental observation. PMID- 15783472 TI - Search for excited and exotic electrons in the egamma decay channel in pp collisions at sqrt[s] = 1.96 TeV. AB - We present a search for excited and exotic electrons (e(*)) decaying to an electron and a photon, both with high transverse momentum. We use 202 pb(-1) of data collected in pp collisions at sqrt[s] = 1.96 TeV with the Collider Detector at Fermilab II detector. No signal above standard model expectation is seen for associated ee(*) production. We discuss the e(*) sensitivity in the parameter space of the excited electron mass M(e(*)) and the compositeness energy scale Lambda. In the contact interaction model, we exclude 132 GeV/c(2)J/psiphi and B(0)(d)-->J/psiK(*0) decays. Lifetimes of the heavy and light mass eigenstates in the B(0)(s) system are separately measured for the first time by determining the relative contributions of amplitudes with definite CP as a function of the decay time. Using 203+/-15 B(0)(s) decays we obtain tau(L) = (1.05(+0.16)(-0.13) +/- 0.02) ps and tau(H) = (2.07(+0.58)(-0.46) +/- 0.03) ps. Expressed in terms of the difference DeltaGamma(s) and average Gamma(s), of the decay rates of the two eigenstates, the results are DeltaGamma(s)/Gamma(s) = (65(+25)(-33) +/- 1)% and DeltaGamma(s) = (0.47(+0.19)(-0.24) +/- 0.01) ps(-1). PMID- 15783497 TI - Charge-order pattern of the low-temperature phase from a monoclinic single domain of NaV2O5 uniquely determined by resonant x-ray scattering. AB - The present resonant x-ray scattering has been performed on a monoclinically split single domain of NaV(2)O(5). The observation of a critically enhanced contrast between V(4+) and V(5+) ions has led us to the unequivocal conclusion of the charge-order pattern of the low-temperature phase of NaV(2)O(5) below T(c) = 35 K. In spite of the possible four types of configuration of the zigzag-type charge-order patterns in the ab plane (A,A',B and B'), the stacking sequence along the c axis is determined as the AAA'A' type by comparison with model calculations. PMID- 15783498 TI - Self-energy correction to momentum-density distribution of positron-electron pairs. AB - Positron two-dimensional angular correlation of annihilation radiation (2D ACAR), i.e., the 2D projection of the electron momentum densities sampled by positron, in Si is employed to verify the prediction of the density functional theory within the local-density approximation (LDA). Carefully conducted test shows that the LDA introduces small but definite discrepancies to the 2D-ACAR anisotropies. Self-energy calculation using the GW method indicates that density-fluctuation contributes anisotropic momentum-density correction and thus improves the agreement between theory and experiment. These results provide valuable annotations to the arguments concerning the accuracy and validity of the LDA and GW schemes. PMID- 15783476 TI - Measurement of the Lambda0b lifetime in the decay lambda0b--> J/psiLambda0 with the D0 detector. AB - We present measurements of the Lambda(0)(b) lifetime in the exclusive decay channel Lambda(0)(b)--> J/psiLambda(0), with J/psi--> mu(+)mu(-) and Lambda(0)--> ppi(-), the B0 lifetime in the decay B0-->J/psiK(0)(S) with J/psi--> mu(+)mu(-) and K(0)(S)-->pi(+)pi(-), and the ratio of these lifetimes. The analysis is based on approximately 250 pb(-1) of data recorded with the D0 detector in pp collisions at sqrt[s] = 1.96 TeV. The Lambda(0)(b) lifetime is determined to be tau(Lambda(0)(b)) = 1.22(+0.22)(-0.18)(stat) +/- 0.04(syst) ps, the B0 lifetime tau(B0) = 1.40(+0.11)(-0.10)(stat) +/- 0.03(syst) ps, and the ratio tau(Lambda(0)(b))/tau(B0) = 0.87(+0.17)(-0.14)(stat) +/- 0.03(syst). In contrast with previous measurements using semileptonic decays, this is the first determination of the Lambda(0)(b) lifetime based on a fully reconstructed decay channel. PMID- 15783499 TI - One-dimensional multiband correlated conductors and Anderson impurity physics. AB - A single Anderson impurity model recently predicted, through its unstable fixed point, the phase diagram of a two-band model correlated conductor, well confirmed by dynamical mean-field theory in infinite dimensions. We study here the one dimensional version of the same model and extract its phase diagram in the opposite limit of reduced dimensionality. As expected for one dimension, the Mott metal-insulator transition at half filling is replaced by a dimerized insulator undimerized Mott insulator transition, while away from half filling the strongly correlated superconductivity for inverted Hund's rule exchange in infinite dimensions is replaced by dominant pairing fluctuations. Many other aspects of the one-dimensional system, in particular, the field theories and their symmetries, are remarkably the same as those of the Anderson impurity, whose importance appears enhanced. PMID- 15783500 TI - Landau quantization effects in the charge-density-wave system (Per)2M(mnt)2 (where M = Au and Pt). AB - A finite transfer integral t(a) orthogonal to the conducting chains of a highly one-dimensional metal gives rise to empty and filled bands that simulate an indirect-gap semiconductor upon formation of a charge-density wave (CDW). In contrast to semiconductors such as Ge and Si with band gaps approximately 1 eV, the CDW system possesses an indirect gap with a greatly reduced energy scale, enabling moderate laboratory magnetic fields to have a major effect. The consequent variation of the thermodynamic gap with magnetic field due to Zeeman splitting and Landau quantization enables the electronic band structure parameters (transfer integrals, Fermi velocity) to be determined accurately. These parameters reveal the orbital quantization limit to be reached at approximately 20 T in (Per)2M(mnt)(2) salts, making them highly unlikely candidates for a recently proposed cascade of field-induced CDW states. PMID- 15783502 TI - Controlled and reproducible domain wall displacement by current pulses injected into ferromagnetic ring structures. AB - In a combined numerical and experimental study, we demonstrate that current pulses of different polarity can reversibly and controllably displace a magnetic domain wall (DW) in submicrometer permalloy (NiFe) ring structures. The critical current densities for DW displacement are correlated with the specific spin structure of the DWs and are compared to results of micromagnetic simulations including a spin-torque term. Using a notch, an attractive local pinning potential is created for the DW resulting in a highly reproducible spin structure of the DW, critical for reliable current-induced switching. PMID- 15783503 TI - Spin Hall current driven by quantum interferences in mesoscopic Rashba rings. AB - We propose an all-electrical nanostructure where pure spin current is induced in the transverse voltage probes attached to a quantum-coherent ballistic one dimensional ring when unpolarized charge current is injected through its longitudinal leads. Tuning of the Rashba spin-orbit coupling in a semiconductor heterostructure hosting the ring generates quasiperiodic oscillations of the predicted spin-Hall current due to spin-sensitive quantum-interference effects caused by the difference in the Aharonov-Casher phase accumulated by opposite spin states. Its amplitude is comparable to that of the spin-Hall current predicted for finite-size (simply connected) two-dimensional electron gases, while it gets reduced gradually in wide two-dimensional rings or due to spin independent disorder. PMID- 15783501 TI - Electron-hole duality and vortex rings in quantum dots. AB - In a quantum-mechanical system, particle-hole duality implies that instead of studying particles, we can get equivalent information by studying the missing particles, the so-called holes. Using this duality picture for fermions in a rotating trap the vortices appear as holes in the Fermi sea. Here we predict that the formation of vortices in quantum dots at high magnetic fields causes oscillations in the energy spectrum which can be experimentally observed using accurate tunneling spectroscopy. We use the duality picture to show that these oscillations are caused by the localization of vortices in rings. PMID- 15783504 TI - Conductance of a perfect thin film with diffuse surface scattering. AB - The conductance of thin films with diffusive surface scattering was solved semiclassically by Fuchs and Sondheimer. However, when the intrinsic electron mean free path is very large or infinite their conductance diverges. In this Letter a simple diffraction picture is presented. It yields a conductance which corresponds to a limiting mean free path of a(2)/lambda(F), where a is the film thickness. PMID- 15783505 TI - Low-temperature susceptibility of the noncentrosymmetric superconductor CePt3Si. AB - We report ac susceptibility measurements of polycrystalline CePt(3)Si down to 60 mK and in applied fields up to 9 T. In a zero applied field, a full Meissner state emerges at temperatures T/T(c) < 0.3, where T(c) = 0.65 K is the onset transition temperature. Though transport measurements show a relatively high upper critical field B(c2) approximately 4-5 T, the low-temperature susceptibility chi(') is quite fragile to the applied field, with chi(') diminishing rapidly in fields of a few kG. Interestingly, the field dependence of chi(') is well described by the power law 4pichi(') + 1 = (B/B(c))(1/2), where B(c) is the field at which the onset of resistance is observed in transport measurements. PMID- 15783506 TI - Pronounced enhancement of the lower critical field and critical current deep in the superconducting state of PrOs4Sb12. AB - We have observed an unexpected enhancement of the lower critical field H(c1)(T) and the critical current I(c)(T) deep in the superconducting state below T approximately 0.6 K (T/T(c) approximately 0.3) in the filled skutterudite heavy fermion superconductor PrOs(4)Sb(12). From a comparison of the behavior of H(c1)(T) with that of the heavy fermion superconductors U(1-x)Th(x)Be(13) and UPt(3), we speculate that the enhancement of H(c1)(T) and I(c)(T) in PrOs(4)Sb(12) reflects a transition into another superconducting phase that occurs below T/T(c) approximately 0.3. An examination of the literature reveals unexplained anomalies in other physical properties of PrOs(4)Sb(12) near T/T(c) approximately 0.3 that correlate with the features we have observed in H(c1)(T) and I(c)(T). PMID- 15783507 TI - Orbital dependence of the fermi liquid state in Sr2RuO4. AB - We have used angle-resolved photoemission spectroscopy to determine the bulk electronic structure of Sr(2)RuO(4) above and below the Fermi liquid crossover near 25 K. Our measurements indicate that the properties of the system are highly orbital dependent. The quasi-2D gamma band displays Fermi liquid behavior while the remaining low energy bands show exotic properties consistent with quasi-1D behavior. In the Fermi liquid state below 25 K, the gamma band dominates the electronic properties, while at higher temperatures the quasi-1D beta and alpha bands become more important. PMID- 15783508 TI - Nonequilibrium quasiparticle relaxation in the vortex state of La2-xSrxCuO4. AB - We have measured the charge dynamics in the vortex state of La(2-x)Sr(x)CuO(4) by femtosecond time-resolved reflectance, which we demonstrate to be a direct probe of low-energy quasiparticle states. Application of a c-axis magnetic field induces regions surrounding vortex cores that display pseudogap charge dynamics. We determine the characteristic width approximately 130 A in optimally doped material and we show that it increases with decreasing doping. These results confirm a new experimental method of probing the microscopic properties of vortices in the cuprates. PMID- 15783509 TI - Spectroscopic evidence for multiple order parameter components in the heavy fermion superconductor CeCoIn5. AB - Point-contact spectroscopy was performed on single crystals of the heavy-fermion superconductor CeCoIn(5) between 150 mK and 2.5 K. A pulsed measurement technique ensured minimal Joule heating over a wide voltage range. The spectra show Andreev reflection characteristics with multiple structures which depend on junction impedance. Spectral analysis using the generalized Blonder-Tinkham-Klapwijk formalism for d-wave pairing revealed two coexisting order parameter components with amplitudes Delta(1) = 0.95 +/- 0.15 meV and Delta(2) = 2.4 +/- 0.3 meV, which evolve differently with temperature. Our observations indicate a highly unconventional pairing mechanism, possibly involving multiple bands. PMID- 15783510 TI - Magnetic fluctuations of stripes in the high temperature cuprate superconductors. AB - Within the time-dependent Gutzwiller approximation for the Hubbard model we compute the magnetic fluctuations of vertical metallic stripes with parameters appropriate for La(1.875)Ba(0.125)CuO(4) (LBCO). For bond- and site-centered stripes the excitation spectra are similar, consisting of a low-energy incommensurate acoustic branch which merges into a "resonance peak" at the antiferromagnetic wave vector and several high-energy optical branches. The acoustic branch is similar to the result of theories assuming localized spins whereas the optical branches are significantly different. Results are in good agreement with a recent inelastic neutron study of LBCO. PMID- 15783511 TI - Effect of vortex-antivortex fluctuations on the heat capacity of a type-II superconducting film. AB - The small size vortex-antivortex pairs proliferation in a type-II superconducting film is considered below T(c). The corresponding contribution to the free energy is calculated. It is shown that these fluctuations give the main temperature dependent contribution to the heat capacity of the superconducting film in the sufficiently large interval of temperatures below the transition point. PMID- 15783512 TI - Possible field-tuned superconductor-insulator transition in high-Tc superconductors: implications for pairing at high magnetic fields. AB - The behavior of some high temperature superconductors (HTSC), such as La(2 x)Sr(x)CuO(4) and Bi(2)Sr(2-x)La(x)CuO(6 + delta), at very high magnetic fields, is similar to that of thin films of amorphous InOx near the magnetic-field-tuned superconductor-insulator transition. Analyzing the InOx data at high fields in terms of persisting local pairing amplitude, we argue by analogy that the local pairing amplitude also persists well into the dissipative state of the HTSCs, the regime commonly denoted as the "normal state" in very high magnetic field experiments. PMID- 15783513 TI - Large magnetic field-induced spectral weight enhancement of high-energy spin excitations in La1.88Sr0.12CuO4. AB - We report electronic Raman scattering experiments on a superconducting La(1.88)Sr(0.12)CuO(4) single crystal in a magnetic field. At low temperatures, the spectral weight of the high-energy two-magnon peak increases linearly with field and is amplified by a factor of more than two at 14 T. The effect disappears at elevated temperatures and is not present in undoped La(2)CuO(4). This observation is discussed in terms of an electronically inhomogeneous state in which the field enhances the volume fraction of a phase with local antiferromagnetic order at the expense of the superconducting phase. PMID- 15783514 TI - Helimagnetism of Fe: high pressure study of an Y2Fe17 single crystal. AB - In this Letter we present direct observation of the Fe helimagnetism in an Y2Fe17 single crystal under pressure. Combined neutron diffraction and magnetization measurements under pressure showed that the collinear ferromagnetic phase of Y2Fe17 is substituted by the pressure induced helical incommensurate phases. The complex pressure-temperature-field behavior of the pressure induced helical magnetic phases is attributed to intrinsic properties of the iron sublattice that gives a valuable contribution to the discussion about dominating theoretical models of magnetism in gamma-Fe. PMID- 15783515 TI - ac-Driven double quantum dots as spin pumps and spin filters. AB - We propose and analyze a new scheme of realizing both spin filtering and spin pumping by using ac-driven double quantum dots in the Coulomb blockade regime. By calculating the current through the system in the sequential tunneling regime, we demonstrate that the spin polarization of the current can be controlled by tuning the parameters (amplitude and frequency) of the ac field. We also discuss spin relaxation and decoherence effects in the pumped current. PMID- 15783516 TI - Direct imaging of asymmetric magnetization reversal in exchange-biased Fe/MnPd bilayers by x-ray photoemission electron microscopy. AB - X-ray photoemission electron microscopy is used to probe the remnant magnetic domain structure in high quality, single-crystalline, exchange-biased Fe/MnPd bilayers. It is found that the induced unidirectional anisotropy strongly affects the overall magnetic domain structure. Real space images of the ferromagnetic domains provide direct evidence for an asymmetric magnetization reversal process after saturation along the ferromagnetic hard direction. The magnetization reversal occurs by moment rotation for decreasing fields while it proceeds by domain nucleation and growth for increasing fields. The observed domains are consistent with the crystallography of the bilayers and favor a configuration that minimizes the overall magnetostatic energy of the ferromagnetic layer. PMID- 15783517 TI - Shape of temperature dependence of spontaneous magnetization of ferromagnets: quantitative analysis. AB - Analysis of available experimental data shows that there exists a limited variety of shapes of temperature dependence of spontaneous magnetization. For most metallic ferromagnets the shape (as opposed to scale) of the M(s) versus T curve can be characterized by a single dimensionless parameter. A numerical description of the dependence M(s)(T) for a particular ferromagnetic material is thus reduced to evaluating three quantities: the saturation magnetization M(0) identical with M(s)(0), the Curie point T(C), and the shape parameter s. It is demonstrated that classical spin (S = infinity) dynamics fails to describe correctly either of the finite-temperature characteristics, T(C) or s. PMID- 15783518 TI - Selective photoswitching of the binuclear spin crossover compound {[Fe(bt)(NCS)2]2(bpm)} into two distinct macroscopic phases. AB - The low-spin (LS-LS, S = 0) diamagnetic form of the binuclear spin crossover complex {[Fe(bt)(NCS)(2)](2)(bpm)} was selectively photoconverted into two distinct macroscopic phases at different excitation wavelengths (1342 or 647.1 nm). These long-lived metastable phases have been identified, respectively, as the symmetry-broken paramagnetic form (HS-LS, S = 2) and the antiferromagnetically coupled (HS-HS, S = 0) high-spin form of the compound. The selectivity may be explained by the strong coupling of the primary excited states to the paramagnetic state. PMID- 15783519 TI - Teleportation of electronic many-qubit states encoded in the electron spin of quantum dots via single photons. AB - We propose a teleportation scheme that relies only on single-photon measurements and Faraday rotation, for teleportation of many-qubit entangled states stored in the electron spins of a quantum dot system. The interaction between a photon and the two electron spins, via Faraday rotation in microcavities, establishes Greenberger-Horne-Zeilinger entanglement in the spin-photon-spin system. The appropriate single-qubit measurements, and the communication of two classical bits, produce teleportation. This scheme provides the essential link between spintronic and photonic quantum information devices by permitting quantum information to be exchanged between them. PMID- 15783520 TI - Chain conformations and photoluminescence of poly(di-n-octylfluorene). AB - The diverse steady-state spectroscopic properties of poly(di-n-octylfluorene) are addressed from a molecular-level perspective. Modeling of representative oligomers support the experimental observation of at least three distinguishable classes of conformational isomers with differing chain torsion angles. One class appears to be populated by a single compact structural isomer, and this conforms to the so-called beta phase. A rigorous Franck-Condon analysis of the photoluminescence in conjunction with Frenkel-type exciton band structure calculations is performed. These results accurately reproduce all major spectral features of the photoabsorption and those of singlet exciton emission. PMID- 15783521 TI - Smearing of phase transition due to a surface effect or a bulk inhomogeneity in ferroelectric nanostructures. AB - The boundary conditions, customarily used in the Landau-type approach to ferroelectric thin films and nanostructures, have to be modified to take into account that a surface of a ferroelectric is a defect of a field type. The surface (interface) field is coupled to a normal component of polarization and, as a result, the second order phase transitions are generally suppressed and anomalies in response are washed out, as observed experimentally. PMID- 15783522 TI - Surface-stimulated nucleation of reverse domains in ferroelectrics. AB - We present a model for reverse domain nucleation in ferroelectrics, which takes into account ferroelectric-electrode coupling in both homogeneous and random cases. The model provides a solution to the coercivity paradox--i.e., the large discrepancy between the observed and predicted coercive fields, common to many systems. We demonstrate the possibility of not thermally activated nucleation of reverse domains. We find that small inhomogeneities in the ferroelectric electrode interface may lead to an exponentially wide spectrum of waiting times for switching. The model predicts that switching is facilitated near morphotropic phase boundaries in perovskite-type ferroelectrics. PMID- 15783523 TI - Enhanced elasticity and soft glassy rheology of a smectic in a random porous environment. AB - We report studies of the frequency-dependent shear modulus, G(*)(omega) = G(')(omega) + iG('')(omega), of the liquid crystal octylcyanobiphenyl (8CB) confined in a colloidal aerosil gel. With the onset of smectic order, G' grows approximately linearly with decreasing temperature, reaching values that exceed by more than 3 orders of magnitude the values for pure 8CB. The modulus at low temperatures possesses a power-law component, G(*)(omega) approximately omega(alpha), with exponent alpha that approaches zero with increasing gel density. The amplitude of G' and its variation with temperature and gel density indicate that the low temperature response is dominated by a dense population of defects in the smectic. In contrast, when the 8CB is isotropic or nematic, the modulus is controlled by the elastic behavior of the colloidal gel. PMID- 15783524 TI - Deuterium NMR investigation of the biaxial nematic phase in an organosiloxane tetrapode. AB - Deuterium NMR is used to examine the molecular order exhibited by an organosiloxane tetrapode giving the first experimental evidence, using a bulk sample, for the existence of a biaxial nematic phase in this type of compounds. The temperature dependence of the averaged quadrupolar coupling constant and asymmetry parameter was determined in the compound's nematic phase. Two distinct regimes could be identified, one with a vanishing asymmetry parameter corresponding to a uniaxial nematic phase and another with a significant temperature dependent asymmetry parameter, corresponding to a biaxial nematic phase. The high values obtained for the asymmetry parameter at the lower end of the nematic range are well above experimental error and constitute a definite proof of the biaxial nature of the nematic phase exhibited by the studied compound for those temperatures. PMID- 15783525 TI - Relation between the high density phase and the very-high density phase of amorphous solid water. AB - It has been suggested that high-density amorphous (HDA) ice is a structurally arrested form of high-density liquid (HDL) water, while low-density amorphous ice is a structurally arrested form of low-density liquid (LDL) water. Recent experiments and simulations have been interpreted to support the possibility of a second distinct high-density structural state, named very high-density amorphous (VHDA) ice, questioning the LDL-HDL hypothesis. We test this interpretation using extensive computer simulations and find that VHDA is a more stable form of HDA and that, in fact, VHDA should be considered as the amorphous ice of the quenched HDL. PMID- 15783526 TI - Universal power-law decay of the impulse energy in granular protectors. AB - Protecting a big impulse from outside is one of the important issues of our everyday life. A granular medium is often used as a protecting material. The impulse inside a granular medium is a solitary wave which may be confined temporarily to a particular region of the medium, which we call the granular container that plays the role of the protector. We find a universal power-law behavior in time for the leakage of the impulse energy confined inside various granular containers. PMID- 15783527 TI - Structure formation in electromagnetically driven granular media. AB - We report structure formation in submonolayers of magnetic microparticles subjected to periodic electrostatic and magnetic excitations. Depending on the excitation parameters, we observe the formation of a rich variety of structures: clusters, rings, chains, and networks. The dynamics and shapes of the structures are strongly dependent on the amplitude and frequency of the external magnetic field. We find that for pure ac magnetic driving the low-frequency magnetic excitation favors compact clusters, whereas high frequency driving favors chains and netlike structures. An abrupt phase transition from chains to a network phase was observed for a high density of particles. PMID- 15783528 TI - Short-range spectroscopic ruler based on a single-molecule optical switch. AB - We demonstrate a novel all-optical switch consisting of two molecules: a primary fluorophore (Cy5) that can be switched between a fluorescent and a dark state by light of different wavelengths, and a secondary chromophore (Cy3) that facilitates switching. The interaction between the two molecules exhibits a distance dependence much steeper than that of conventional Fo rster resonance energy transfer. This enables the switch to act as a ruler with the capability to probe distances difficult to access by other spectroscopic methods, thus presenting a new tool for the study of biomolecules at the single-molecule level. PMID- 15783529 TI - Filament depolymerization by motor molecules. AB - Motor proteins that specifically interact with the ends of cytoskeletal filaments can induce filament depolymerization. A phenomenological description of this process is presented. We show that under certain conditions motors dynamically accumulate at the filament ends. We compare simulations of two microscopic models to the phenomenological description. The depolymerization rate can exhibit maxima and dynamic instabilities as a function of the bulk motor density for processive depolymerization. We discuss our results in relation to experimental studies of Kin-13 family motor proteins. PMID- 15783530 TI - Preference of sensory neural coding for 1/f signals. AB - We investigated the influences of different types of temporal correlations in the input signal on the functions and coding properties of neurons in the primary visual cortex (V1). We found that the temporal transfer functions of V1 neurons exhibit higher gain, and the spike responses exhibit higher coding efficiency and information transmission rates, for the 1/f (natural long-term correlation) signals than for 1/f(0) (no correlation) and 1/f(2) (stronger long-term correlation) signals. These results suggest that the intermediate long-term correlation ubiquitous to natural signals may play an important role in shaping and optimizing the machinery of neurons in their adaptation to the natural environment. PMID- 15783531 TI - Theory of mitotic spindle oscillations. AB - During unequal cell division the mitotic spindle is positioned away from the center of the cell before cell cleavage. In many biological systems this repositioning is accompanied by oscillatory movements of the spindle. We present a theoretical description for mitotic spindle oscillations. We show that the cooperative attachment and detachment of cortical force generators to astral microtubules leads to spontaneous oscillations beyond a critical number of force generators. This mechanism can quantitatively describe the spindle oscillations observed during unequal division of the one cell stage Caenorhabditis elegans embryo. PMID- 15783532 TI - Earthquakes descaled: on waiting time distributions and scaling laws. AB - Recently, several authors have used waiting time distributions for large earthquake data sets to draw conclusions regarding the physics of earthquake processes. We show, theoretically and by simulation, that a characteristic kink in observed waiting time distributions does not have the physical significance of separating correlated and uncorrelated earthquakes. It also follows from our discussion that the Omori law is not trivially related to a proposed scaling law and that caution must be taken before the spatial scaling exponent of the law is interpreted as a fractal dimension of seismicity. PMID- 15783533 TI - Quantum turbulence in condensate collisions: an application of the classical field method. AB - We apply the classical field method to simulate the production of correlated atoms during the collision of two Bose-Einstein condensates. Our nonperturbative method includes the effect of quantum noise, and describes collisions of high density condensates with very large out-scattered fractions. Quantum correlation functions for the scattered atoms show that the correlation between pairs of atoms of opposite momentum is rather small. We also predict the existence of quantum turbulence in the field of the scattered atoms. PMID- 15783534 TI - Quantum glassiness in strongly correlated clean systems: an example of topological overprotection. AB - This Letter presents solvable examples of quantum many-body Hamiltonians of systems that are unable to reach their ground states as the environment temperature is lowered to absolute zero. These examples, three-dimensional generalizations of quantum Hamiltonians proposed for topological quantum computing, (1) have no quenched disorder, (2) have solely local interactions, (3) have an exactly solvable spectrum, (4) have topologically ordered ground states, and (5) have slow dynamical relaxation rates akin to those of strong structural glasses. PMID- 15783535 TI - Observation of hybrid soliton vortex-ring structures in Bose-Einstein condensates. AB - We present the experimental discovery of compound structures comprising solitons and vortex rings in Bose-Einstein condensates. We examine both their creation via soliton-vortex collisions and their subsequent development, which is largely governed by the dynamics of interacting vortex rings. A theoretical model in three-dimensional cylindrical symmetry is also presented. PMID- 15783536 TI - Atomic quantum dots coupled to a reservoir of a superfluid Bose-Einstein condensate. AB - We study the dynamics of an atomic quantum dot, i.e., a single atom in a tight optical trap which is coupled to a superfluid reservoir via laser transitions. Quantum interference between the collisional interactions and the laser induced coupling results in a tunable dot-bath coupling, allowing an essentially complete decoupling from the environment. Quantum dots embedded in a 1D Luttinger liquid of cold bosonic atoms realize a spin-boson model with Ohmic coupling, which exhibits a dissipative phase transition and allows us to directly measure atomic Luttinger parameters. PMID- 15783537 TI - Coherent molecular optics using ultracold sodium dimers. AB - Coherent molecular optics is performed using two-photon Bragg scattering. Molecules were produced by sweeping an atomic Bose-Einstein condensate through a Feshbach resonance. The spectral width of the molecular Bragg resonance corresponded to an instantaneous temperature of 20 nK, indicating that atomic coherence was transferred directly to the molecules. An autocorrelating interference technique was used to observe the quadratic spatial dependence of the phase of an expanding molecular cloud. Finally, atoms initially prepared in two momentum states were observed to cross pair with one another, forming molecules in a third momentum state. This process is analogous to sum-frequency generation in optics. PMID- 15783538 TI - Experimental quantum coin tossing. AB - In this Letter we present the first implementation of a quantum coin-tossing protocol. This protocol belongs to a class of "two-party" cryptographic problems, where the communication partners distrust each other. As with a number of such two-party protocols, the best implementation of the quantum coin tossing requires qutrits, resulting in a higher security than using qubits. In this way, we have also performed the first complete quantum communication protocol with qutrits. In our experiment the two partners succeeded to remotely toss a row of coins using photons entangled in the orbital angular momentum. We also show the experimental bounds of a possible cheater and the ways of detecting him. PMID- 15783539 TI - Noiseless quantum circuits for the Peres separability criterion. AB - In this Letter we give a method for constructing sets of simple circuits that can determine the spectrum of a partially transposed density matrix, without requiring either a tomographically complete positive-operator-valued measurement or the addition of noise to make the spectrum non-negative. These circuits depend only on the dimension of the Hilbert space and are otherwise independent of the state. PMID- 15783540 TI - Unconditional security of a three state quantum key distribution protocol. AB - Quantum key distribution (QKD) protocols are cryptographic techniques with security based only on the laws of quantum mechanics. Two prominent QKD schemes are the Bennett-Brassard 1984 and Bennett 1992 protocols that use four and two quantum states, respectively. In 2000, Phoenix et al. proposed a new family of three-state protocols that offers advantages over the previous schemes. Until now, an error rate threshold for security of the symmetric trine spherical code QKD protocol has been shown only for the trivial intercept-resend eavesdropping strategy. In this Letter, we prove the unconditional security of the trine spherical code QKD protocol, demonstrating its security up to a bit error rate of 9.81%. We also discuss how this proof applies to a version of the trine spherical code QKD protocol where the error rate is evaluated from the number of inconclusive events. PMID- 15783541 TI - Quantum nonlocality obtained from local states by entanglement purification. AB - We have applied an entanglement purification protocol to produce a single entangled pair of photons capable of violating a Clauser-Horne-Shimony-Holt Bell inequality from two pairs that individually could not. The initial poorly entangled photons were created by a controllable decoherence that introduced complex errors. All of the states were reconstructed using quantum state tomography which allowed for a quantitative description of the improvement of the state after purification. PMID- 15783542 TI - Experimental quantum cloning with prior partial information. AB - When prior partial information about a state to be cloned is available, it can be cloned with a fidelity higher than that of universal quantum cloning. We experimentally verify this intriguing relationship between the cloning fidelity and the prior information by reporting the first experimental optimal quantum state-dependent cloner, using nuclear magnetic resonance techniques. Our experiments may further cast important implications into many quantum information processing protocols. PMID- 15783543 TI - Robust two-qubit quantum registers. AB - We carry out a systematic analysis of a pair of coupled qubits, each of which is subject to its own dissipative environment, and argue that a combination of the interqubit couplings which provides for the lowest possible decoherence rates corresponds to the incidence of a double spectral degeneracy in the two-qubit system. We support this general argument by the results of an evolutionary genetic algorithm which can also be used for optimizing time-dependent processes (gates) and their sequences that implement various quantum computing protocols. PMID- 15783544 TI - Fault-tolerant quantum computation via exchange interactions. AB - Quantum computation can be performed by encoding logical qubits into the states of two or more physical qubits, and control of effective exchange interactions and possibly a global magnetic field. This "encoded universality" paradigm offers potential simplifications in quantum computer design since it does away with the need to control physical qubits individually. Here we show how encoded universality schemes can be combined with fault-tolerant quantum error correction, thus establishing the scalability of such schemes. PMID- 15783545 TI - Detection and characterization of structural changes in the hard-disk fluid under freezing and melting conditions. AB - The fluid of two-dimensional hard disks is investigated over a range of densities by Monte Carlo simulations in order to detect and characterize structural changes which take place when the condition of freezing and melting is approached. A novel method is proposed based on the use of the Voronoi tessellation and a certain shape factor which turns out to be a clear indicator of the presence of different underlying substructures (domains). Close to the freezing condition the probability distribution of the shape factor develops a second distinct maximum corresponding to a predominant presence of near-regular hexagons, whereas the original peak, having its origin primarily in pentagons and distorted hexagons, diminishes and disappears at melting density. PMID- 15783546 TI - Test of charge conjugation invariance. AB - We report on the first determination of upper limits on the branching ratio (BR) of eta decay to pi0pi0gamma and to pi0pi0pi0gamma. Both decay modes are strictly forbidden by charge conjugation (C) invariance. Using the Crystal Ball multiphoton detector, we obtained BR(eta-->pi0pi0gamma)<5 x 10(-4) at the 90% confidence level, in support of C invariance of isoscalar electromagnetic interactions of the light quarks. We have also measured BR(eta- >pi0pi0pi0gamma)<6 x 10(-5) at the 90% confidence level, in support of C invariance of isovector electromagnetic interactions. PMID- 15783551 TI - Order-to-chaos transition in rotational nuclei. AB - We have developed a new method to study the order-to-chaos transition in rotational nuclei. Correlations between successive gamma rays are used to determine the average complexity of the intermediate levels and thereby the ratio of the interaction potential between levels to the level spacing. The measured ratios, 0.15 to 1.5, span the range from nearly fully ordered to nearly fully chaotic. PMID- 15783552 TI - Reconciling coulomb dissociation and radiative capture measurements. AB - We calculate the energy spectrum for 8B dissociation on a Pb target to all orders in the Coulomb and nuclear fields, and show that the slope of S17(E) obtained in previous analyses of Coulomb dissociation data is too steep, due to deficiencies in the conventional first-order analysis that was used. With a more complete theory that avoids the far-field approximation and includes E2, nuclear and dynamical projectile polarization, the disagreement between indirect and direct methods for determining the S17(E) slope and the extrapolated S17(0) values is reduced significantly. PMID- 15783553 TI - Production of doubly charged helium ions by two-photon absorption of an intense sub-10-fs soft x-ray pulse at 42 eV photon energy. AB - We report on the observation of doubly charged helium ions produced by a nonlinear interaction between a helium atom and photons with a photon energy of 42 eV which are generated with the 27th harmonic of a femtosecond pulse from a Ti:sapphire laser. The number of ions is proportional to the square of the intensity of the 27th harmonic pulse, and thus two-photon double ionization should be dominantly induced as compared with other nonlinear processes accompanying sequential ionization via a singly charged ion. This phenomenon is utilized to measure the pulse duration of the 27th harmonic pulse by using an autocorrelation technique, for the first time to our knowledge, and as a result a duration of 8 fs is found. PMID- 15783554 TI - Density functional theory with correct long-range asymptotic behavior. AB - We derive an exact representation of the exchange-correlation energy within density functional theory (DFT) which spawns a class of approximations leading to correct long-range asymptotic behavior. Using a simple approximation, we develop an electronic structure theory that combines a new local correlation energy (based on Monte Carlo calculations applied to the homogeneous electron gas) and a combination of local and explicit long-ranged exchange. The theory is applied to several first-row atoms and diatomic molecules where encouraging results are obtained: good description of the chemical bond at the same time allowing for bound anions, reasonably accurate affinity energies, and correct polarizability of an elongated hydrogen chain. Further stringent tests of DFT are passed, concerning ionization potential and charge distribution under large bias. PMID- 15783555 TI - Fragmentation of water by heavy ions. AB - Absolute cross sections for fragmentation of water molecules by C3+ and O5+ ions over an energy region where the Bragg peak maximizes were measured for ionization, electron capture, and electron loss channels. A collision regime where sigmaSigmaOq+> or =sigmaH2O+ was reached for the first time, producing large abundances of H+ and O+ fragments in comparison to proton impact. Our findings have straightforward implications in the subsequent fast chemistry at the ionization site and on the O production in the first stages of water radiolysis. An unexpected channel-independent relationship between the cross sections for the fragmentation products, which is also approximately independent of the particle type, energy, and charge state, is found. A model is presented to explain such behavior allowing the cross sections of all fragmentation products to be obtained from single and double electron removal cross sections. PMID- 15783556 TI - Strongly coupled matter field and nonanalytic decay rate of dipole molecules in a waveguide. AB - The decay rate gamma of an excited dipole molecule inside a waveguide is evaluated for the strongly coupled matter-field case near a cutoff frequency omegac without using perturbation analysis. Because of the singularity in the density of photon states at the cutoff frequency, we find that gamma depends nonanalytically on the coupling constant g as g4/3, which leads to a vast increase in the decay rate. PMID- 15783547 TI - Search for supersymmetry with gauge-mediated breaking in diphoton events at D0. AB - We report the results of a search for supersymmetry (SUSY) with gauge-mediated breaking in the missing transverse energy distribution of inclusive diphoton events using 263 pb(-1) of data collected by the D0 experiment at the Fermilab Tevatron Collider in 2002-2004. No excess is observed above the background expected from standard model processes, and lower limits on the masses of the lightest neutralino and chargino of about 108 and 195 GeV, respectively, are set at the 95% confidence level. These are the most stringent limits to date for models with gauge-mediated SUSY breaking with a short-lived neutralino as the next-to-lightest SUSY particle. PMID- 15783557 TI - Nonlinear interactions with an ultrahigh flux of broadband entangled photons. AB - We experimentally demonstrate sum-frequency generation with entangled photon pairs, generating as many as 40,000 photons per second, visible even to the naked eye. The nonclassical nature of the interaction is exhibited by a linear intensity dependence of the nonlinear process. The key element in our scheme is the generation of an ultrahigh flux of entangled photons while maintaining their nonclassical properties. This is made possible by generating the down-converted photons as broadband as possible, orders of magnitude wider than the pump. This approach can be applied to other nonlinear interactions, and may become useful for various quantum-measurement tasks. PMID- 15783558 TI - Coherent inelastic backscattering of intense laser light by cold atoms. AB - We present a nonperturbative treatment of coherent backscattering of intense laser light from cold atoms and predict a nonvanishing backscattering signal even at very large intensities, due to the constructive (self-)interference of inelastically scattered photons. PMID- 15783559 TI - Near-field radiation from a ridge waveguide transducer in the vicinity of a solid immersion lens. AB - A near-field optical system is investigated to improve the transmission efficiency of near-field transducers. A ridge waveguide is placed adjacent to a solid immersion lens (SIL) but separated by a low-index dielectric layer. The incident electric field near the focus of the SIL is determined by the Richards Wolf vector field equations. The finite element method is used to solve Maxwell's equations. A spot size of 31 nm is obtained. The maximum value of the absorbed optical power density in the recording medium is 7.51 x 10(-4) mW/nm3 for a 100 mW input power. PMID- 15783560 TI - Stable ring-profile vortex solitons in bessel optical lattices. AB - Stable ring-profile vortex solitons, featuring a bright shape, appear to be very rare in nature. However, here we show that they exist and can be made dynamically stable in defocusing cubic nonlinear media with an imprinted Bessel optical lattice. We find the families of vortex solitons and reveal their salient properties, including the conditions required for their stability. We show that the higher the soliton topological charge, the deeper the lattice modulation necessary for stabilization. PMID- 15783548 TI - Measurements of the branching fraction and CP-violation asymmetries in B0- >f0(980)K0S. AB - We present measurements of the branching fraction and CP-violating asymmetries in the decay B0-->f0(980)K0S. The results are obtained from a data sample of 123 x 10(6) Upsilon(4S)-->BB decays. From a time-dependent maximum likelihood fit, we measure the branching fraction B(B0-->f0(980)(-->pi+pi-)K0)=(6.0+/-0.9+/-0.6+/ 1.2)x10(-6), the mixing-induced CP violation parameter S=-1.62(+0.56)(-0.51)+/ 0.09+/-0.04, and the direct CP violation parameter C=0.27+/-0.36+/-0.10+/-0.07, where the first errors are statistical, the second systematic, and the third due to model uncertainties. We measure the f0(980) mass and width to be mf0(980)=(980.6+/-4.1+/-0.5+/-4.0) MeV/c2 and Gammaf0(980)=(43(+12)(-9)+/-3+/-9) MeV/c2, respectively. PMID- 15783561 TI - Quantum freeze of fidelity decay for chaotic dynamics. AB - We show that the mechanism of quantum freeze of fidelity decay for perturbations with a zero time average, recently discovered for a specific case of integrable dynamics [New J. Phys. 5, 109 (2003)], can be generalized to arbitrary quantum dynamics. We work out explicitly the case of a chaotic classical counterpart, for which we find semiclassical expressions for the value and the range of the plateau of fidelity. After the plateau ends, we find explicit expressions for the asymptotic decay, which can be exponential or Gaussian depending on the ratio of the Heisenberg time to the decay time. Arbitrary initial states can be considered; e.g., we discuss coherent states and random states. PMID- 15783562 TI - Drag reduction in bubbly Taylor-Couette turbulence. AB - In Taylor-Couette flow the total energy dissipation rate and therefore the drag can be determined by measuring the torque on the system. We do so for Reynolds numbers between Re=7 x 10(4) and Re=10(6) after having injected (i) small bubbles (R=1 mm) up to a volume concentration of alpha=5% and (ii) buoyant particles (rhop/rhol=0.14) of comparable volume concentration. In case (i) we observe a crossover from little drag reduction at smaller Re to strong drag reduction up to 20% at Re=10(6). In case (ii) we observe at most little drag reduction throughout. Several theoretical models for bubbly drag reduction are discussed in view of our findings. PMID- 15783565 TI - Reaching the nonlinear regime of Raman amplification of ultrashort laser pulses. AB - The intensity of a subpicosecond laser pulse was amplified by a factor of up to 1000 using the Raman backscatter interaction in a 2 mm long gas jet plasma. The process of Raman amplification reached the nonlinear regime, with the intensity of the amplified pulse exceeding that of the pump pulse by more than an order of magnitude. Features unique to the nonlinear regime such as gain saturation, bandwidth broadening, and pulse shortening were observed. Simulation and theory are in qualitative agreement with the measurements. PMID- 15783564 TI - Experimental benchmark for an improved simulation of absolute soft-x-ray emission from polystyrene targets irradiated with the Nike laser. AB - Absolutely calibrated, time-resolved spectral intensity measurements of soft-x ray emission (hnu approximately 0.1-1.0 keV) from laser-irradiated polystyrene targets are compared to radiation-hydrodynamic simulations that include our new postprocessor, Virtual Spectro. This new capability allows a unified, detailed treatment of atomic physics and radiative transfer in nonlocal thermodynamic equilibrium conditions for simple spectra from low-Z materials as well as complex spectra from high-Z materials. The excellent agreement (within a factor of approximately 1.5) demonstrates the powerful predictive capability of the codes for the complex conditions in the ablating plasma. A comparison to data with high spectral resolution (E/deltaE approximately 1000) emphasizes the importance of including radiation coupling in the quantitative simulation of emission spectra. PMID- 15783563 TI - Wave spectra in solid and liquid complex (dusty) plasmas. AB - Spectra of longitudinal and transverse waves were obtained experimentally in liquid and solid two-dimensional complex (dusty) plasmas at different kinetic temperatures. As the temperature increased and the phase state of the plasma changed from solid to liquid, the phonon spectra of both longitudinal and transverse modes broadened (especially at high wave numbers), indicating increased damping. The transverse mode disappeared and a thermal (compressional) mode appeared. PMID- 15783566 TI - Comparison of laser ion acceleration from the front and rear surfaces of thin foils. AB - The comparative efficiency and beam characteristics of high-energy ions generated by high-intensity short-pulse lasers (approximately 1-6 x 10(19) W/cm2) from both the front and rear surfaces of thin metal foils have been measured under identical conditions. Using direct beam measurements and nuclear activation techniques, we find that rear-surface acceleration produces higher energy particles with smaller divergence and a higher efficiency than front-surface acceleration. Our observations are well reproduced by realistic particle-in-cell simulations, and we predict optimal criteria for future applications. PMID- 15783567 TI - Creep ruptures in heterogeneous materials. AB - We present creep experiments on fiber composite materials with different controlled heterogeneity. All samples exhibit a power-law relaxation of the strain rate in the primary creep regime (Andrade's law) followed by a power-law acceleration up to rupture. We discover that the rupture time is proportional to the duration of the primary creep regime, showing the interplay between the two regimes and offering a method of rupture prediction. These experimental results are rationalized by a mean-field model of representative elements with nonlinear viscoelastic rheology and with a large heterogeneity of strengths. PMID- 15783568 TI - Direct imaging of the alkali metal site in K-doped fullerene peapods. AB - The structure of K-doped fullerene peapods has been investigated by means of high resolution transmission electron microscopy and electron energy-loss spectroscopy (EELS). It is proven that the potassium atoms can be doped at the intermolecular sites within C60 peapods. The EELS spectrum of potassium (K) L edge clearly exhibits the feature of K+ in the doped peapod and consequently suggests n-type doping. These results encourage the realization of a one-dimensional superconducting wire based on the nanotube peapods. PMID- 15783569 TI - Almost compact breathers in anharmonic lattices near the continuum limit. AB - Certain strictly anharmonic one-dimensional lattices support discrete breathers over a macroscopic localized domain that in the continuum limit becomes exactly compact. The discrete breather tails decay at a double-exponential rate, so such systems can store energy locally, especially since discrete breathers appear to be stable for amplitudes below a sharp stability threshold. The effective width of other solutions broadens over time, but, under appropriate conditions, only after a positive waiting time. The continuum limit of a planar hexagonal lattice also supports a compact breather. PMID- 15783549 TI - Measurement of Wgamma and Zgamma production in pp collisions at square root s=1.96 TeV. AB - The standard model predictions for Wgamma and Zgamma production are tested using an integrated luminosity of 200 pb(-1) of pp collision data collected at the Collider Detector at Fermilab. The cross sections are measured by selecting leptonic decays of the W and Z bosons, and photons with transverse energy ET>7 GeV that are well separated from leptons. The production cross sections and kinematic distributions for the Wgamma and Zgamma data are compared to SM predictions. PMID- 15783550 TI - Measurement of the B0s lifetime in the exclusive decay channel B0s-->J/psiphi. AB - Using the exclusive decay B0s-->J/psi(mu+mu-)phi(K+K-), we report the most precise single measurement of the B0s lifetime. The data sample corresponds to an integrated luminosity of approximately 220 pb(-1) collected with the D0 detector at the Fermilab Tevatron Collider in 2002-2004. We reconstruct 337 signal candidates, from which we extract the B0s lifetime, tau(B0s)=1.444(+0.098)( 0.090)(stat)+/-0.020(sys) ps. We also report a measurement for the lifetime of the B0 meson using the exclusive decay B0-->J/psi(mu+mu-)K*0(892)(K+pi-). We reconstruct 1370 signal candidates, obtaining tau(B0)=1.473(+0.052)( 0.050)(stat)+/-0.023(sys) ps, and the ratio of lifetimes, tau(B0s)/tau(B0)=0.980(+0.076)(-0.071)(stat)+/-0.003(sys). PMID- 15783570 TI - Atomic correlation between adjacent graphene layers in double-wall carbon nanotubes. AB - Atomic correlation between adjacent graphene layers was elucidated for double wall carbon nanotubes (DWNTs) through a chiral index assignment of two nested nanotubes by high-resolution transmission electron microscopy. Our analysis provides a rather constant diameter difference close to 0.75 nm but no chiral angle correlation between the constituent nanotubes in the concentric DWNTs. The local atomic correlation as a commensurate graphene stacking was repeatedly found in eccentric DWNTs and circumscribed nanotubes, which should lead to elastic deformation and bundling of nanotubes. PMID- 15783573 TI - First principles calculation of the interdiffusion coefficient in binary alloys. AB - The atomic mechanisms of diffusion in alloys are complex due to the variations of migration energies with environment and the correlations induced by short-range order between the different components. We present a first-principles approach for calculating vacancy-mediated diffusion coefficients in crystalline binary alloys and apply it to obtain the interdiffusion coefficient of Al(1-x)Lix. The rigorous treatment of atomic migration indicates that short- and long-range order induces strongly correlated migration mechanisms that deviate from random walk behavior. PMID- 15783572 TI - XY spin fluid in an external magnetic field. AB - A method of integral equations is developed to study anisotropic fluids with planar spins in an external field. As a result, the calculations for these systems appear to be no more difficult than those for ordinary homogeneous liquids. The approach proposed is applied to the ferromagnetic XY spin fluid in a magnetic field using a soft mean spherical closure and the Born-Green-Yvon equation. This provides an accurate reproduction of the complicated phase diagram behavior obtained by cumbersome Gibbs ensemble simulation and multiple histogram reweighting techniques. PMID- 15783571 TI - Hydrogen-adsorption induced atomic rearrangement of a Pb monolayer on Si(111). AB - We have observed interesting H-atom adsorption induced atomic rearrangements of a Pb monolayer on the Si(111) with a scanning tunneling microscope. A hexagonal ringlike pattern is formed around the point defect. The interactions among nearby H-adsorbed defects can even produce interferencelike superstructures. Phase boundaries are found to either enhance or suppress the formation of the interference pattern. These phenomena are produced by an intricate interplay between electronic and atomic interactions as perturbed by the adsorbed hydrogen atoms. PMID- 15783574 TI - Intrinsic character of the (3 x 3) to (square root 3 x square root 3) phase transition in Pb/Si(111). AB - We have investigated the (3 x 3) to (square root 3 x square root 3) reversible phase transition in Pb/Si(111) by means of variable temperature scanning tunneling microscopy and density functional first-principles calculations. By tracking exactly the same regions of the surface with atomic resolution in a temperature range between 40 and 200 K, we have observed the phase transition in real time. The ability to prepare and track exceptionally large domains without defects has allowed us to detect the intrinsic character of the phase transition at temperatures around 86 K. This intrinsic character is in full agreement with our first-principles calculations. Moreover, our results show that the hypothesis that point defects play a fundamental role as the driving force, reported for similar systems, can be discarded for Pb/Si(111). PMID- 15783575 TI - New information on water interfacial structure revealed by phase-sensitive surface spectroscopy. AB - A phase-sensitive sum-frequency vibrational spectroscopic technique is developed to study interfacial water structure of water/quartz interfaces. Measurements allow deduction of both real and imaginary parts of the surface nonlinear spectral response, revealing an unprecedentedly detailed picture of the net polar orientations of the water species at the interface. The orientations of the icelike and liquidlike species appear to respond very differently to the bulk pH change indicating the existence of different surface sites on quartz with different deprotonation pK values. PMID- 15783576 TI - Selecting the shape of supported metal nanocrystals: Pd huts, hexagons, or pyramids on SrTiO3(001). AB - Increasing interest in oxide supported nanoparticle science and technology is stimulating research into controlling nanocrystal shape. Pd forms nanocrystals on the surface of SrTiO3(001), and depending on the crystallographic interface of the Pd with the substrate three shapes can be created: truncated pyramids, huts, and hexagonal shaped disks. Scanning tunneling microscopy reveals that the nanocrystal shapes are determined by the substrate reconstruction and the substrate temperature during deposition. A thermodynamic model is used to show that the pyramids and huts are stable structures, and that the hexagons are trapped in a metastable state. PMID- 15783577 TI - Non-Fermi-liquid behavior within the ferromagnetic phase in URu2-xRexSi2. AB - The URu2-xRexSi2 system exhibits ferromagnetic order for Re concentrations 0.3 < x < or =1.0. Non-Fermi-liquid (NFL) behavior is observed in the specific heat for 0.15< or = x< or =0.6 [C/T proportional to, -lnT (or T(-0.1))], and also in the power-law T dependence of the electrical resistivity [rhoT proportional to, Tn] with n<2 for 0.15< or = x <0.8, at low T, providing strong evidence that the NFL behavior persists within the ferromagnetic phase. Furthermore, the deviation of the physical properties of URu2-xRexSi2 from Fermi-liquid behavior is most pronounced at the ferromagnetic quantum critical point, and the NFL behavior found in the ferromagnetic phase may be consistent with the Griffiths-McCoy phase model. PMID- 15783578 TI - Nonlinear screening theory of the Coulomb glass. AB - A nonlinear screening theory is formulated to study the problem of gap formation and its relation to glassy freezing in classical Coulomb glasses. We find that a pseudogap ("plasma dip") in a single-particle density of states begins to open already at temperatures comparable to the Coulomb energy. This phenomenon is shown to reflect the emergence of short-range correlations in a liquid (plasma) phase, a process which occurs even in the absence of disorder. Glassy ordering emerges when disorder is present, but this occurs only at temperatures roughly an order of magnitude lower. Our result demonstrate that the formation of the plasma dip at high temperatures is a process distinct from the formation of the Efros Shklovskii pseudogap, which in our model emerges only within the glassy phase. PMID- 15783579 TI - Magnetic interactions influence the properties of helium defects in iron. AB - Density functional theory calculations of He defect properties in iron have shown an unexpected influence of magnetism arising from the defect's electronic structure. In contrast with previous work that neglected such effects, the results indicate that the tetrahedral position is energetically more favorable for the He interstitial than the octahedral site. This may have significant implications for He clustering and bubble nucleation, which will impact material performance in future fusion reactors. These results provide the basis for development of improved atomistic models. PMID- 15783580 TI - Quantum critical behavior near a density-wave instability in an isotropic fermi liquid. AB - We study the quantum critical behavior in an isotropic Fermi liquid in the vicinity of a zero-temperature density-wave transition at a finite wave vector qc. We show that, near the transition, the Landau damping of the soft bosonic mode yields a crossover in the fermionic self-energy from Sigma(k,omega) approximately Sigma(k) to Sigma(k,omega) approximately Sigma(omega), where k and omega are momentum and frequency. Because of this self-generated locality, the fermionic effective mass diverges right at the quantum critical point, not before; i.e., the Fermi liquid survives up to the critical point. PMID- 15783581 TI - Hybrid phase at the quantum melting of the Wigner crystal. AB - We study the quantum melting of the two-dimensional Wigner crystal using a fixed node quantum Monte Carlo approach. In addition to the two already known phases (Fermi liquid at large density and Wigner crystal at low density), we find a third stable phase at intermediate values of the density. The third phase has hybrid behaviors in between a liquid and a solid. This hybrid phase has the nodal structure of a Slater determinant constructed out of the bands of a triangular lattice. PMID- 15783582 TI - Thermopower of interacting GaAs bilayer hole systems in the reentrant insulating phase near nu=1. AB - We report thermopower measurements of interacting GaAs bilayer hole systems. When the carrier densities in the two layers are equal, these systems exhibit a reentrant insulating phase near the quantum Hall state at total filling factor nu=1. Our data show that, as the temperature is decreased, the thermopower diverges in the insulating phase. This behavior indicates the opening of an energy gap at low temperature, consistent with the formation of a pinned Wigner solid. We extract an energy gap and a Wigner solid melting phase diagram. PMID- 15783583 TI - Superconductivity in CeCoIn5-xSnx: veil over an ordered state or novel quantum critical point? AB - Measurements of specific heat and electrical resistivity in magnetic fields up to 9 T along [001] and temperatures down to 50 mK of Sn-substituted CeCoIn5 are reported. The maximal -ln(T) divergence of the specific heat at the upper critical field Hc2 down to the lowest temperature characteristic of non-Fermi liquid systems at the quantum critical point (QCP), the universal scaling of the Sommerfeld coefficient, and agreement of the data with spin-fluctuation theory provide strong evidence for quantum criticality at Hc2 for all x< or =0.12 in CeCoIn5-xSnx. These results indicate the "accidental" coincidence of the QCP located near Hc2 in pure CeCoIn5, in actuality, constitute a novel quantum critical point associated with unconventional superconductivity. PMID- 15783584 TI - Creep motion of an elastic string in a random potential. AB - We study the creep motion of an elastic string in a two-dimensional pinning landscape by means of Langevin dynamics simulations. We find that the velocity force characteristics are well described by the creep formula predicted from phenomenological scaling arguments. We analyze the creep exponent mu and the roughness exponent zeta. Two regimes are identified: when the temperature is larger than the strength of the disorder, we find mu approximately 1/4 and zeta approximately 2/3, in agreement with the quasi-equilibrium-nucleation picture of creep motion; on the contrary, when lowering the temperature enough, the values of mu and zeta increase, showing a strong violation of the latter picture. PMID- 15783585 TI - Anomalous vortex motion in the quantum-liquid phase of amorphous MoxSi1-x films. AB - We measure, in real time (t), the fluctuating component of the flux-flow voltage V(t), deltaV(t) identical withV(t)-V0, about the average V0 in the vortex-liquid phase of amorphous MoxSi1-x films. For the thick film, deltaV(t) originating from the vortex motion is clearly visible in the quantum-liquid phase, where the distribution of deltaV(t) is asymmetric, indicative of large velocity and/or number fluctuations of driven vortices. For the thin film the similar anomalous vortex motion is observed in nearly the same (reduced-)temperature regime. These results suggest that vortex dynamics in the low-temperature liquid phase of thick and thin films is dominated by common physical mechanisms, presumably related to quantum effects. PMID- 15783586 TI - Half-filled layered organic superconductors and the resonating-valence-bond theory of the hubbard-heisenberg model. AB - We present a resonating-valence-bond theory of superconductivity for the Hubbard Heisenberg model on an anisotropic triangular lattice. Our calculations are consistent with the observed phase diagram of the half-filled layered organic superconductors, such as the beta, beta', kappa, and lambda phases of (BEDT TTF)2X [bis(ethylenedithio)tetrathiafulvalene] and (BETS)2X [bis(ethylenedithio)tetraselenafulvalene]. We find a first order transition from a Mott insulator to a dx2-y2 superconductor with a small superfluid stiffness and a pseudogap with dx2-y2 symmetry. PMID- 15783587 TI - Angle-resolved photoemission spectroscopy of the antiferromagnetic superconductor Nd1.87Ce0.13CuO4: anisotropic spin-correlation gap, pseudogap, and the induced quasiparticle mass enhancement. AB - We performed high-resolution angle-resolved photoemission spectroscopy on Nd1.87Ce0.13CuO4, which is located at the boundary of the antiferromagnetic (AF) and the superconducting phase. We observed that the quasiparticle (QP) effective mass around (pi,0) is strongly enhanced due to the opening of the AF gap. The QP mass and the AF gap are found to be anisotropic, with the largest value near the intersecting point of the Fermi surface and the AF zone boundary. In addition, we observed that the QP peak disappears around the Neel temperature (TN) while the AF pseudogap is gradually filled up at much higher temperatures, possibly due to the short-range AF correlation. PMID- 15783588 TI - Critical point and the nature of the pseudogap of single-layered copper-oxide Bi2Sr2-xLaxCuO6+delta superconductors. AB - We apply strong magnetic fields of H=28.5 to 43 T to suppress superconductivity (SC) in the cuprates Bi2Sr2-xLaxCuO6+delta (x=0.65, 0.40, 0.25, 0.15, and 0), and investigate the low temperature (T) normal state by 63Cu nuclear spin-lattice relaxation rate (1/T1) measurements. We find that the pseudogap (PG) phase persists deep inside the overdoped region but terminates at x approximately 0.05, which corresponds to the hole doping concentration of approximately 0.21. Beyond this critical point, the normal state is a Fermi liquid that persists as the ground state when superconductivity is removed by the magnetic field. A comparison of the superconducting state with the H-induced normal state in the x=0.40 (Tc=32 K) sample indicates that there remains substantial part of the Fermi surface even in the fully developed PG state, which suggests that the PG and SC are coexisting matters. PMID- 15783589 TI - Phase diagram and critical exponents of a dissipative Ising spin chain in a transverse magnetic field. AB - We consider a one-dimensional Ising model in a transverse magnetic field coupled to a dissipative heat bath. The phase diagram and the critical exponents are determined from extensive Monte Carlo simulations. It is shown that the character of the quantum phase transition is radically altered from the corresponding nondissipative model and the double well coupled to a dissipative heat bath with linear friction. Spatial couplings and the dissipative dynamics combine to form a new quantum criticality which is independent of dissipation strength. PMID- 15783590 TI - Magnetic-field induced transition to the 1/2 magnetization plateau state in the geometrically frustrated magnet CdCr2O4. AB - The magnetization of the geometrically frustrated spinel CdCr2O4 was measured in pulsed fields of up to 47 T. We found a metamagnetic transition to a very wide magnetization plateau state with one half of the full moment of S=3/2 Cr3+ at 28 T, independent of the field direction. This is the first observation of magnetization plateau state realized in Heisenberg pyrochlore magnet. The plateau state can be ascribed to a collinear spin configuration with three-up and one down spins out of four spins of each Cr tetrahedron. A large magnetostriction is observed at the transition in spite of the negligible spin-orbit couplings. We argue that spin frustration plays a vital role in this large spin-lattice coupling. PMID- 15783591 TI - d-d excitations in manganites probed by resonant inelastic x-ray scattering. AB - We report a study of electronic excitations in manganites exhibiting a range of ground states, using resonant inelastic x-ray scattering (RIXS) at the Mn K edge. Excitations with temperature dependent changes correlated with the magnetism were observed as high as 10 eV. By calculating Wannier functions, and finite-q response functions, we associate this dependence with intersite d-d excitations. The calculated dynamical structure factor is found to be similar to the RIXS spectra. PMID- 15783592 TI - Experimental observation of the spin-Hall effect in a two-dimensional spin-orbit coupled semiconductor system. AB - We report the experimental observation of the spin-Hall effect in a 2D hole system with spin-orbit coupling. The 2D hole layer is a part of a p-n junction light-emitting diode with a specially designed coplanar geometry which allows an angle-resolved polarization detection at opposite edges of the 2D hole system. In equilibrium the angular momenta of the spin-orbit split heavy-hole states lie in the plane of the 2D layer. When an electric field is applied across the hole channel, a nonzero out-of-plane component of the angular momentum is detected whose sign depends on the sign of the electric field and is opposite for the two edges. Microscopic quantum transport calculations show only a weak effect of disorder, suggesting that the clean limit spin-Hall conductance description (intrinsic spin-Hall effect) might apply to our system. PMID- 15783593 TI - Single-beam and enhanced two-beam second-harmonic generation from silicon nanocrystals by use of spatially inhomogeneous femtosecond pulses. AB - Optical second-harmonic generation (SHG) is used as a noninvasive probe of the interfaces of Si nanocrystals (NCs) embedded uniformly in an SiO2 matrix. Measurements of the generated SH mode verify that the second-harmonic polarization has a nonlocal dipole form proportional to (E x Delta inverted) E that depends on inhomogeneities in the incident field E, as proposed in recent models based on a locally noncentrosymmetric dipolar response averaged over the spherical NC interfaces. A two-beam SHG geometry is found to enhance this polarization greatly compared to single-beam SHG, yielding strong signals useful for scanning, spectroscopy, and real-time monitoring. This configuration provides a general strategy for enhancing the second-order nonlinear response of centrosymmetric samples, as demonstrated here for both Si nanocomposites and their glass substrates. PMID- 15783594 TI - Optical pumping of the electronic and nuclear spin of single charge-tunable quantum dots. AB - We present a comprehensive examination of optical pumping of spins in individual GaAs quantum dots as we change the net charge from positive to neutral to negative with a charge-tunable heterostructure. Negative photoluminescence polarization memory is enhanced by optical pumping of ground state electron spins, which we prove with the first measurements of the Hanle effect on an individual quantum dot. We use the Overhauser effect in a high longitudinal magnetic field to demonstrate efficient optical pumping of nuclear spins for all three charge states of the quantum dot. PMID- 15783595 TI - Optical absorption and photoluminescence spectroscopy of the growth of silver nanoparticles. AB - Results obtained from the optical absorption and photoluminescence (PL) spectroscopy experiments have shown the formation of excitons in the silver exchanged glass samples. These findings are reported here for the first time. Further, we investigate the dramatic changes in the photoemission properties of the silver-exchanged glass samples as a function of postannealing temperature. Observed changes are thought to be due to the structural rearrangements of silver and oxygen bonding during the heat treatments of the glass matrix. In fact, photoelectron spectroscopy does reveal these chemical transformations of silver exchanged soda glass samples caused by the thermal effects of annealing in a high vacuum atmosphere. An important correlation between temperature-induced changes of the PL intensity and thermal growth of the silver nanoparticles has been established in this Letter through precise spectroscopic studies. PMID- 15783596 TI - Large optical nonlinearity of semiconducting single-walled carbon nanotubes under resonant excitations. AB - We measured third-order nonlinear susceptibility (chi(3)) spectra in semiconducting single-walled carbon nanotubes (SWNTs) by the Z-scan method. |Imchi(3)| is remarkably enhanced under resonant excitation to the lowest interband transition, reaching 4.2 x 10(-6) esu and 1.5 x 10(-7) esu in SWNTs grown by the laser ablation and HiPco methods, respectively. A comparison of the transient absorption changes evaluated by degenerate and nondegenerate pump-probe measurements suggests that the resonant enhancement of |Imchi(3)| is dominated by a coherent process rather than by saturation of absorption. PMID- 15783597 TI - Domain-enhanced interlayer coupling in ferroelectric/paraelectric superlattices. AB - We investigate the ferroelectric phase transition and domain formation in a periodic superlattice consisting of alternate ferroelectric (FE) and paraelectric (PE) layers of nanometric thickness. We find that the polarization domains formed in the different FE layers can interact with each other via the PE layers. By coupling the electrostatic equations with those obtained by minimizing the Ginzburg-Landau functional, we calculate the critical temperature of transition Tc as a function of the FE/PE superlattice wavelength Lambda and quantitatively explain the recent experimental observation of a thickness dependence of the ferroelectric transition temperature in KTaO3/KNbO3 strained-layer superlattices. PMID- 15783598 TI - Texture in the superconducting order parameter of CeCoIn5 revealed by nuclear magnetic resonance. AB - We present a 115In NMR study of the quasi-two-dimensional heavy-fermion superconductor CeCoIn5 believed to host a Fulde-Ferrel-Larkin-Ovchinnikov (FFLO) state. In the vicinity of the upper critical field and with a magnetic field applied parallel to the ab plane, the NMR spectrum exhibits a dramatic change below T*(H) which well coincides with the position of reported anomalies in specific heat and ultrasound velocity. We argue that our results provide the first microscopic evidence for the occurrence of a spatially modulated superconducting order parameter expected in a FFLO state. The NMR spectrum also implies an anomalous electronic structure of vortex cores. PMID- 15783599 TI - Ferroelectricity and tetragonality in ultrathin PbTiO3 films. AB - The evolution of tetragonality with thickness has been probed in epitaxial c-axis oriented PbTiO3 films with thicknesses ranging from 500 down to 24 A. High resolution x ray pointed out a systematic decrease of the c-axis lattice parameter with decreasing film thickness below 200 A. Using a first-principles model Hamiltonian approach, the decrease in tetragonality is related to a reduction of the polarization attributed to the presence of a residual unscreened depolarizing field. It is shown that films below 50 A display a significantly reduced polarization but still remain ferroelectric. PMID- 15783600 TI - Temperature dependence of isotopic quantum effects in water. AB - The technique of high energy x-ray diffraction has been used to measure the temperature variation of hydrogen versus deuterium isotopic quantum effects on the structure of water. The magnitude of the effect is found to be inversely proportional to the temperature, varying by a factor of 2.5 over the range 6 to 45 degrees C. In addition, the H216O versus H218O effect has been measured at 26 degrees C and the structural difference shown to be restricted to the nearest neighbor molecular interactions. The results are compared to recent simulations and previously measured isochoric temperature differentials; additionally, implications for H/D substitution experiments are considered. PMID- 15783602 TI - Subdiffusive axial transport of granular materials in a long drum mixer. AB - Granular mixtures segregate radially by size when tumbled in a partially filled horizontal drum. The smaller component moves toward the axis of rotation and forms a buried core, which then splits into axial bands. Models have generally assumed that the axial segregation is opposed by diffusion. Using narrow pulses of the smaller component as initial conditions, we have characterized axial transport in the core. We find that the axial advance of the segregated core is well described by a self-similar concentration profile whose width scales as talpha, with alpha approximately 0.3<1/2. Thus, the process is subdiffusive rather than diffusive as previously assumed. We compare our results to two one dimensional model equations which contain self-similarity and subdiffusion: a linear fractional diffusion model and the nonlinear porous medium equation. PMID- 15783603 TI - Instantaneous velocity profiles during granular avalanches. AB - We perform experimental measurements of the instantaneous velocity profile of the flowing layer during granular avalanches. In the pile depth, the velocity profile follows a pure exponential decrease in contrast with steady flows that are known to exhibit a well developed upper linear part. The velocity profile in the pile width is a plug flow with two exponential boundary layers at the walls. Even though no steady state is observed during the avalanche, these velocity profiles are self-similar and build up almost instantaneously, with time independent characteristic lengths. PMID- 15783601 TI - Shock-wave theory for rupture of rubber. AB - This Letter presents a theory for the rupture of rubber. Unlike conventional cracks, ruptures in rubber travel faster than the speed of sound and consist of two oblique shocks that meet at a point. Physical features of rubber needed for this phenomenon include Kelvin dissipation and an increase of toughness as rubber retracts. There are three levels of theoretical description: an approximate continuum theory, an exact analytical solution of a slightly simplified discrete problem, and numerical solution of realistic and fully nonlinear equations of motion. PMID- 15783604 TI - Transition between tonic spiking and bursting in a neuron model via the blue-sky catastrophe. AB - We study a continuous and reversible transition between periodic tonic spiking and bursting activities in a neuron model. It is described as the blue-sky catastrophe, which is a homoclinic bifurcation of a saddle-node periodic orbit of codimension one. This transition constitutes a biophysically plausible mechanism for the regulation of burst duration that increases with no bound like 1/square root alpha-alpha0 as the transition value alpha0 is approached. PMID- 15783605 TI - Asymmetric diffusion through synthetic nanopores. AB - We show that diffusion currents for a membrane containing a single conical nanopore with a fixed surface charge and small enough opening diameter depend on the concentration gradient direction. We interpret the results based on the effect of salt concentration on the thickness of the electrical double layer within the nanopore associated with the nanopore's surface charge and the distribution of electric fields inside the pore. The experimental observations are described by a diffusional model based on the Smoluchowski-Nernst-Planck equation. PMID- 15783606 TI - Highly stretched single polymers: atomic-force-microscope experiments versus ab initio theory. AB - Experimental single-molecule stretching curves for three backbone architectures (single-stranded DNA, various types of peptides, polyvinylamine) are quantitatively compared with corresponding quantum-chemical (zero-temperature) ab initio calculations in the high-force range of up to two nanonewtons. For high forces, quantitative agreement is obtained with the contour length of the polymers as the only fitting parameter. For smaller forces, the effects of chain fluctuations are accounted for by using recent theoretical results for the stretching response of a freely-rotating-chain model. PMID- 15783607 TI - Manning-Oosawa counterion condensation. AB - Counterion condensation is a basic feature of 2D electrostatics exhibited by highly charged rodlike polymers such as DNA. In the framework of the Poisson Boltzmann equation with salt, we show that such a polymer of radius a attracts a condensate of thickness RM=A(axi)1/2 where xi is the Debye length and A depends weakly on the polymer charge density q0. To leading order in 1/ln(xi/a), we derive the condensate structure and show that free ions follow universal density profiles independent of a and q0. Generalizing this approach we calculate ion profiles for finite concentration solutions. PMID- 15783608 TI - Analysis of the spatial distribution between successive earthquakes. AB - Spatial distances between subsequent earthquakes in southern California exhibit scale-free statistics, with a critical exponent delta approximately 0.6, as well as finite size scaling. The statistics are independent of the threshold magnitude as long as the catalog is complete, but depend strongly on the temporal ordering of events, rather than the geometry of the spatial epicenter distribution. Nevertheless, the spatial distance and waiting time between subsequent earthquakes are uncorrelated with each other. These observations contradict the theory of aftershock zone scaling with main shock magnitude. PMID- 15783609 TI - Long-term memory: a natural mechanism for the clustering of extreme events and anomalous residual times in climate records. AB - We study the statistics of the return intervals between extreme events above a certain threshold in long-term persistent records. We find that the long-term memory leads (i) to a stretched exponential distribution of the return intervals, (ii) to a pronounced clustering of extreme events, and (iii) to an anomalous behavior of the mean residual time to the next event that depends on the history and increases with the elapsed time in a counterintuitive way. We present an analytical scaling approach and demonstrate that all these features can be seen in long climate records. The phenomena should also occur in heartbeat records, Internet traffic, and stock market volatility and have to be taken into account for an efficient risk evaluation. PMID- 15783610 TI - Comment on "secure communication using mesoscopic coherent states". PMID- 15783612 TI - Comment on "photoproduction of eta-mesic 3He". PMID- 15783614 TI - Measurement of geometric phase for mixed states using single photon interferometry. AB - Geometric phase may enable inherently fault-tolerant quantum computation. However, due to potential decoherence effects, it is important to understand how such phases arise for mixed input states. We report the first experiment to measure mixed-state geometric phases in optics, using a Mach-Zehnder interferometer, and polarization mixed states that are produced in two different ways: decohering pure states with birefringent elements; and producing a nonmaximally entangled state of two photons and tracing over one of them, a form of remote state preparation. PMID- 15783615 TI - Dynamics of a bright soliton in Bose-Einstein condensates with time-dependent atomic scattering length in an expulsive parabolic potential. AB - We present a family of exact solutions of the one-dimensional nonlinear Schro dinger equation which describes the dynamics of a bright soliton in Bose-Einstein condensates with the time-dependent interatomic interaction in an expulsive parabolic potential. Our results show that, under a safe range of parameters, the bright soliton can be compressed into very high local matter densities by increasing the absolute value of the atomic scattering length, which can provide an experimental tool for investigating the range of validity of the one dimensional Gross-Pitaevskii equation. We also find that the number of atoms in the bright soliton keeps dynamic stability: a time-periodic atomic exchange is formed between the bright soliton and the background. PMID- 15783617 TI - Decay rate of an excited atom in a moving medium. AB - The effect of a uniformly moving medium on the decay rate of an excited atom is calculated. When the local field effects are neglected, the free space decay rate is modified by a factor which is a complicated function of refractive index, permeability, and velocity of the medium. It is shown that the decay rate vanishes if the velocity of the medium exceeds the phase velocity of light in the stationary medium. PMID- 15783618 TI - Entanglement in quantum spin chains, symmetry classes of random matrices, and conformal field theory. AB - We compute the entropy of entanglement between the first N spins and the rest of the system in the ground states of a general class of quantum spin chains. We show that under certain conditions the entropy can be expressed in terms of averages over ensembles of random matrices. These averages can be evaluated, allowing us to prove that at critical points the entropy grows like kappalog(2N+kappa as N-->infinity, where kappa and kappa are determined explicitly. In an important class of systems, kappa is equal to one-third of the central charge of an associated Virasoro algebra. Our expression for kappa therefore provides an explicit formula for the central charge. PMID- 15783616 TI - BCS-BEC crossover in a gas of Fermi atoms with a p-wave feshbach resonance. AB - We investigate unconventional superfluidity in a gas of Fermi atoms with an anisotropic p-wave Feshbach resonance. Including the p-wave Feshbach resonance as well as the associated three kinds of quasimolecules with finite orbital angular momenta Lz=+/-1,0, we calculate the transition temperature of the superfluid phase. As one passes through the p-wave Feshbach resonance, we find the usual BCS BEC crossover phenomenon. The p-wave BCS state continuously changes into the BEC of bound molecules with L=1. Our calculation includes the effect of fluctuations associated with Cooper pairs and molecules which are not Bose condensed. PMID- 15783620 TI - Provably secure experimental quantum bit-string generation. AB - Coin tossing is a cryptographic task in which two parties who do not trust each other aim to generate a common random bit. Using classical communication this is impossible, but nontrivial coin tossing is possible using quantum communication. Here we consider the case when the parties do not want to toss a single coin, but many. This is called bit-string generation. We report the experimental generation of strings of coins which are provably more random than achievable using classical communication. The experiment is based on the "plug and play" scheme developed for quantum cryptography, and therefore well suited for long distance quantum communication. PMID- 15783619 TI - Teleportation of an atomic ensemble quantum state. AB - We propose a protocol to achieve high fidelity quantum state teleportation of a macroscopic atomic ensemble using a pair of quantum-correlated atomic ensembles. We show how to prepare this pair of ensembles using quasiperfect quantum state transfer processes between light and atoms. Our protocol relies on optical joint measurements of the atomic ensemble states and magnetic feedback reconstruction. PMID- 15783621 TI - Detecting vacuum entanglement in a linear ion trap. AB - We propose and study a method for detecting ground-state entanglement in a chain of trapped ions. We show that the entanglement between single ions or groups of ions can be swapped to the internal levels of two ions by sending laser pulses that couple the internal and motional degrees of freedom. This allows us to entangle two ions without actually performing gate operations. A proof of principle of the effect can be realized with two trapped ions and is feasible with current technology. PMID- 15783622 TI - Calculation of partition functions by measuring component distributions. AB - A new algorithm is presented, which allows us to calculate numerically the partition function Z for systems, which can be described by arbitrary interaction graphs and lattices, e.g., Ising models or Potts models (for arbitrary values q>0), including random or diluted models. The new approach is suitable for large systems. The basic idea is to measure the distribution of the number of connected components in the corresponding Fortuin-Kasteleyn representation and to compare with the case of zero degrees of freedom, where the exact result Z=1 is known. As an application, d=2 and d=3 dimensional ferromagnetic Potts models are studied, and the critical values qc, where the transition changes from second to first order, are determined. Large systems of sizes N=1000(2) and N=100(3) are treated. The critical value qc(d=2)=4 is confirmed and qc(d=3)=2.35(5) is found. PMID- 15783623 TI - Diffusion-reaction in branched structures: theory and application to the lung acinus. AB - An exact "branch by branch" calculation of the diffusional flux is proposed for partially absorbed random walks on arbitrary tree structures. In the particular case of symmetric trees, an explicit analytical expression is found which is valid whatever the size of the tree. Its application to the respiratory phenomena in pulmonary acini gives an analytical description of the crossover regime governing the human lung efficiency. PMID- 15783624 TI - Optical clocks based on ultranarrow three-photon resonances in alkaline Earth atoms. AB - A sharp resonance line that appears in three-photon transitions between the 1S0 and 3P0 states of alkaline earth and Yb atoms is proposed as an optical frequency standard. This proposal permits the use of the even isotopes, in which the clock transition is narrower than in proposed clocks using the odd isotopes and the energy interval is not affected by external magnetic fields or the polarization of trapping light. With this method, the width and the rate of the clock transition can, in principle, be continuously adjusted from the MHz level to sub mHz without loss of signal amplitude by varying the intensities of the three optical beams. Doppler and recoil effects can be eliminated by proper alignment of the three optical beams or by point confinement in a lattice trap. Light-shift effects on the clock accuracy can be limited to below a part in 10(18). PMID- 15783625 TI - Asymptotic silence of generic cosmological singularities. AB - In this Letter we investigate the nature of generic cosmological singularities using the framework developed by Uggla et al. We study the past asymptotic dynamics of general vacuum G2 cosmologies, which are expected to capture the singular behavior of generic cosmologies with no symmetries at all. Our results indicate that asymptotic silence holds, i.e., particle horizons along all time lines shrink to zero for generic solutions. Moreover, we provide evidence that spatial derivatives become dynamically insignificant along generic time lines, and that the evolution into the past along such time lines is governed by an asymptotic dynamical system which is associated with an invariant set-the silent boundary. We identify an attracting subset on the silent boundary that organizes the oscillatory dynamics of generic time lines in the singular regime. Finally, we discuss the dynamics associated with recurring spike formation. PMID- 15783626 TI - Origin of the binary pulsar J0737-3039B. AB - Evolutionary scenarios suggest that the progenitor of the new binary pulsar J0737 3039B was a He star with M>(2.1-2.3)M. . We show that this case implies that the binary must have a large (>120 km/s) center of mass velocity. However, the location, approximately 50 pc from the Galactic plane, suggests that the system has, at high likelihood, a significantly smaller center of mass velocity and a progenitor more massive than 2.1M. is ruled out (at 97% C.L.). A progenitor mass around 1.45M. , involving a new previously unseen gravitational collapse, is kinematically favored. The low mass progenitor is consistent with the recent scintillation based velocity measurement of 66+/-15 km/s and rules out the high mass solution at 99% C.L. Conversely, if the unlikely higher mass solution is the true one we should increase the estimated rate of neutron star mergers by a factor of at least 2. PMID- 15783627 TI - Dark energy reflections in the redshift-space quadrupole. AB - We show that the redshift-space quadrupole will be a powerful tool for constraining dark energy even if the baryon oscillations are missing from the monopole power spectrum and bias is scale and time dependent. We calculate the accuracy with which next-generation galaxy surveys such as KAOS will measure the quadrupole power spectrum, which gives the leading anisotropies in the power spectrum in redshift space due to linear velocity, and the so-called "Finger of God" and Alcock-Paczynski effects. Combining the monopole and quadrupole power spectra, in the complete absence of baryon oscillations (Omegab=0), leads to a roughly 500% improvement in constraints on dark energy compared with those from the monopole spectrum alone. PMID- 15783628 TI - Large matter effects in numu-->nutau oscillations. AB - We show that matter effects change the numu-->nutau oscillation probability by as much as 70% for certain ranges of energies and path lengths. Consequently, the numu-->nutau survival probability also undergoes large changes. A proper understanding of numu survival rates must consider matter effects in Pmutau as well as Pmue. We comment on (a) how these matter effects may be observed and the sign of Delta31 determined in atmospheric neutrino measurements and at neutrino factories, and (b) how they lead to heightened sensitivity for small theta13. PMID- 15783630 TI - Discrete and continuum spectra in the unified shell model approach. AB - A new version of the nuclear shell model unifies the consideration of the discrete spectrum, where the results reproduce the standard shell model, and continuum. The ingredients of the method are the non-Hermitian effective Hamiltonian, energy-dependent one-body and two-body decay amplitudes, and self consistent treatment of thresholds. The results for helium and oxygen isotope chains reproduce the data well. PMID- 15783631 TI - Production of the neutron-rich hypernucleus 10LambdaLi in the (pi-,K+) double charge-exchange reaction. AB - In order to produce a neutron-rich Lambda hypernucleus for the first time, we carried out an experiment by utilizing the (pi-,K+) double charge-exchange reaction on a 10B target. We observed the production of a 10LambdaLi hypernucleus. The cross section for the Lambda bound region was found to be 11.3+/-1.9 nb/sr with the 1.2 GeV/c incident momentum, which is compared with the 10LambdaB hypernucleus production cross section, 7.8+/-0.3 microb/sr, in the (pi+,K+) reaction with a 1.05 GeV/c incident momentum beam. PMID- 15783629 TI - System-size dependence of strangeness production in nucleus-nucleus collisions at squareroot[sNN]=17.3 GeV. AB - Emission of pi+/-, K+/-, phi, and Lambda was measured in near-central C+C and Si+Si collisions at 158 AGeV beam energy. Together with earlier data for p+p, S+S, and Pb+Pb, the system-size dependence of relative strangeness production in nucleus-nucleus collisions is obtained. Its fast rise and the saturation observed at about 60 participating nucleons can be understood as the onset of the formation of coherent systems of increasing size. PMID- 15783632 TI - Neutron multiplicity in the fission of 238U and 235U with neutrons up to 200 MeV. AB - Prompt-fission-neutron multiplicities were measured for 238U(n,f) and 235U(n,f) from 0.4 to 200 MeV. The data are of great importance in connection with accelerator-coupled nuclear reactor systems incinerating actinides. We report that fission induced by 200 MeV neutrons produces approximately 10 more prompt neutrons than fission induced by reactor neutrons. Most neutrons are evaporated from the fission fragments and the prefission compound nucleus, as the preequilibrium emission of energetic neutrons accounts for a maximum of 15% of the prompt neutrons at 200 MeV. PMID- 15783633 TI - Sympathetic cooling of 4He+ ions in a radio-frequency trap. AB - We have generated Coulomb crystals of ultracold 4He+ ions in a linear radio frequency trap, by sympathetic cooling via laser-cooled 9Be+. Stable crystals containing up to 150 localized He+ ions at approximately 20 mK were obtained. Ensembles or single ultracold He+ ions open up interesting perspectives for performing precision tests of QED and measurements of nuclear radii. This Letter also indicates the feasibility of cooling and crystallizing highly charged atomic ions using 9Be+ as coolant. PMID- 15783634 TI - Dynamic Jahn-Teller effects in isolated C60- studied by near-infrared spectroscopy in a storage ring. AB - We have measured the near-infrared absorption spectrum for isolated C60- ions at room temperature. Two bands, at 9145 cm(-1) and 10460 cm(-1), have been identified in addition to the main absorption band at 9382 cm(-1), seen also at low temperature in a matrix. An interpretation based on the theory of dynamic Jahn-Teller effects is proposed. PMID- 15783635 TI - Compression of atomic phase space using an asymmetric one-way barrier. AB - We show how to construct asymmetric optical barriers for atoms. These barriers can be used to compress phase-space of a sample by creating a confined region in space where atoms can accumulate with heating at the single photon recoil level. We illustrate our method with a simple two-level model and then show how it can be applied to more realistic multilevel atoms. PMID- 15783636 TI - Attosecond probing of vibrational dynamics with high-harmonic generation. AB - The numerical solution of the time-dependent Schrodinger equation for vibrating hydrogen molecules in few-cycle laser pulses shows that high-harmonic generation is sensitive to the laser-induced vibrational motion. More intense harmonics are generated in heavier isotopes, the difference increasing with the harmonic frequency. Analytical theory reveals a dependence of the harmonics on the vibrational autocorrelation function. With the help of a genetic algorithm, the nuclear motion can be reconstructed from the harmonic spectra with sub-fs time resolution. PMID- 15783637 TI - Optical-fiber source of polarization-entangled photons in the 1550 nm telecom band. AB - We present a fiber-based source of polarization-entangled photons that is well suited for quantum communication applications in the 1550 nm band of standard fiber-optic telecommunications. Polarization entanglement is created by pumping a nonlinear-fiber Sagnac interferometer with two time-delayed orthogonally polarized pump pulses and subsequently removing the time distinguishability by passing the parametrically scattered signal and idler photon pairs through a piece of birefringent fiber. Coincidence detection of the signal and idler photons yields biphoton interference with visibility greater than 90%, while no interference is observed in direct detection of either signal or idler photons. All four Bell states can be prepared with our setup and we demonstrate violations of the Clauser-Horne-Shimony-Holt form of Bell's inequality by up to 10 standard deviations of measurement uncertainty. PMID- 15783638 TI - Core relaxation in atomic ultrastrong laser field ionization. AB - We have investigated atomic ionization dynamics in Kr in the transition regime from nonrelativistic to relativistic laser intensities (10(16) to 10(18) W/cm2) by measuring yields of highly charged ions stemming from an inner shell. Interpretation of the data is focused on the applicability of the single active electron description, which requires fully relaxed core states between successive ionization steps. In particular, we are concerned with transient core polarization or alignment effects originating from the strong dependence of the ionization rates on the magnetic quantum number. We found that for intense laser pulses with 40 fs pulse width internal m-mixing processes appear to be sufficiently fast to erase any transient core polarization. PMID- 15783639 TI - Entangled and disentangled evolution for a single atom in a driven cavity. AB - For an atom in an externally driven cavity, we show that special initial states lead to near-disentangled atom-field evolution, and superpositions of these can lead to near maximally entangled states. Somewhat counterintutively, we find that (moderate) spontaneous emission in this system actually leads to a transient increase in entanglement beyond the steady-state value. We also show that a particular field correlation function could be used, in an experimental setting, to track the time evolution of this entanglement. PMID- 15783640 TI - Transition from antibunching to bunching in cavity QED. AB - The photon statistics of the light emitted from an atomic ensemble into a single field mode of an optical cavity is investigated as a function of the number of atoms. The light is produced in a Raman transition driven by a pump laser and the cavity vacuum, and a recycling laser is employed to repeat this process continuously. For weak driving, a smooth transition from antibunching to bunching is found for about one intracavity atom. Remarkably, the bunching peak develops within the antibunching dip. The observed behavior is well explained by a model describing an ensemble of independent emitters. PMID- 15783641 TI - Plasmon-assisted two-slit transmission: Young's experiment revisited. AB - We present an experimental and theoretical study of the optical transmission of a thin metal screen perforated by two subwavelength slits, separated by many optical wavelengths. The total intensity of the far-field double-slit pattern is shown to be reduced or enhanced as a function of the wavelength of the incident light beam. This modulation is attributed to an interference phenomenon at each of the slits, instead of at the detector. The interference arises as a consequence of the excitation of surface plasmons propagating from one slit to the other. PMID- 15783642 TI - Fast causal information transmission in a medium with a slow group velocity. AB - It is widely believed that the velocity of information upsiloni encoded on an optical pulse is equal to the group velocity upsilong, at least when upsilong is less than the speed of light in vacuum c. On the other hand, several authors suggest that upsiloni=c, although the size of the signal traveling at this velocity may be small, thereby making it difficult to measure. Here, we measure upsiloni for pulses propagating through a resonant "slow-light" medium where upsilong approximately 0.006c. We find upsiloni=1.03c(+0.49c)-0.25c, or that upsiloni approximately 168upsilong, clearly demonstrating that the speed of information cannot be generally described by upsilong, but is characterized by its own velocity. PMID- 15783643 TI - Signatures of homoclinic motion in quantum chaos. AB - Homoclinic motion plays a key role in the organization of classical chaos in Hamiltonian systems. In this Letter, we show that it also imprints a clear signature in the corresponding quantum spectra. By numerically studying the fluctuations of the widths of wave functions localized along periodic orbits we reveal the existence of an oscillatory behavior that is explained solely in terms of the primary homoclinic motion. Furthermore, our results indicate that it survives the semiclassical limit. PMID- 15783644 TI - Flooding of chaotic eigenstates into regular phase space islands. AB - We introduce a criterion for the existence of regular states in systems with a mixed phase space. If this condition is not fulfilled chaotic eigenstates substantially extend into a regular island. Wave packets started in the chaotic sea progressively flood the island. The extent of flooding by eigenstates and wave packets increases logarithmically with the size of the chaotic sea and the time, respectively. This new effect is observed for the example of island chains with just ten islands. PMID- 15783645 TI - Hamiltonian chaos acts like a finite energy reservoir: accuracy of the Fokker Planck approximation. AB - The Hamiltonian dynamics of slow variables coupled to fast degrees of freedom is modeled by an effective stochastic differential equation. Formal perturbation expansions, involving a Markov approximation, yield a Fokker-Planck equation in the slow subspace which respects conservation of energy. A detailed numerical and analytical analysis of suitable model systems demonstrates the feasibility of obtaining the system specific drift and diffusion terms and the accuracy of the stochastic approximation on all time scales. Non-Markovian and non-Gaussian features of the fast variables are negligible. PMID- 15783646 TI - Dynamic van der waals theory of two-phase fluids in heat flow. AB - We present a dynamic van der Waals theory. It is useful to study phase separation when the temperature varies in space. We show that, if heat flow is applied to liquid suspending a gas droplet at zero gravity, a convective flow occurs such that the temperature gradient within the droplet nearly vanishes. As the heat flux is increased, the droplet becomes attached to the heated wall that is wetted by liquid in equilibrium. In one case corresponding to partial wetting by gas, an apparent contact angle can be defined. In the other case with larger heat flux, the droplet completely wets the heated wall expelling liquid. PMID- 15783647 TI - Dynamics of two-sign point vortices in positive and negative temperature states. AB - Dynamics of two-sign point vortices in two-dimensional circular boundary is examined by numerical simulations with MDGRAPE-2. The vortex system is characterized by the inverse temperature beta as determined from the density of states of the microcanonical ensemble of numerically generated 10(7) states. The massive simulation shows that different configurations appear in the time asymptotic state depending on the sign of beta. Condensation of the same-sign vortices is observed when beta<0, while the both-sign vortices tend to be uniformly neutralized when beta>0. During the condensation, a part of the vortices gains energy to form clumps (patches), and the other part of the vortices loses energy to keep the total energy constant and mixes with vortices of the other sign. This observation demonstrates a characteristic feature of negative beta states that the system energy concentrates into the clumps of the same-sign vortices. PMID- 15783648 TI - Four sided domains in hierarchical space dividing patterns. AB - The cracks observed in the glaze of ceramics form networks, which divide the 2D plane into domains. It is shown that, on the average, the number of sides of these domains is four. This contrasts with the usual 2D space divisions observed in Voronoi tessellation or 2D soap froths. In the latter networks, the number of sides of a domain coincides with the number of its neighbors, which, according to Euler's theorem, has to be six on average. The four sided property observed in cracks is the result of a formation process which can be understood as the successive divisions of domains with no later reorganization. It is generic for all networks having such hierarchical construction rules. We introduce a "geometrical charge," analogous to Euler's topological charge, as the difference from four of the number of sides of a domain. It is preserved during the pattern formation of the crack pattern. PMID- 15783649 TI - Structure-based interpretation of the Strouhal-Reynolds number relationship. AB - The wake in the flow past a circular cylinder has posed a long-standing challenge to scientists since the late 19th century. Many aspects of this seemingly simple phenomenon remain unexplained. Of particular interest is the relationship between the dimensionless vortex shedding frequency (the Strouhal number St) and the ratio of inertial to viscous forces in the fluid (the Reynolds number Re). We propose a new St-Re relation based on the observations of the structure of a vortex street in flowing soap films. The measurements suggest a simple two parameter form St=1/(A+B/Re) that describes laminar vortex shedding remarkably well for bulk fluids as well as for two-dimensional flowing soap films. PMID- 15783650 TI - Observation of vertical betatron sideband due to electron clouds in the KEKB low energy ring. AB - The effects of electron clouds on positively charged beams have been an active area of research in recent years at particle accelerators around the world. Transverse beam-size blowup due to electron clouds has been observed in some machines and is considered to be a major limiting factor in the development of higher-current, higher-luminosity electron-positron colliders. The leading proposed mechanism for beam blowup is the excitation of a fast head-tail instability due to short-range wakes within the electron cloud. We present here observations of betatron oscillation sidebands in bunch-by-bunch spectra that may provide direct evidence of such head-tail motion in a positron beam. PMID- 15783651 TI - Focusing hard x rays to nanometer dimensions by adiabatically focusing lenses. AB - We address the question of what is the smallest spot size that hard x rays can be focused to using refractive optics. A thick refractive x-ray lens is considered, whose aperture is gradually (adiabatically) adapted to the size of the beam as it converges to the focus. These adiabatically focusing lenses are shown to have a relatively large numerical aperture, focusing hard x rays down to a lateral size of 2 nm (FWHM), well below the theoretical limit for focusing with waveguides [C. Bergemann, Phys. Rev. Lett.912003204801]. PMID- 15783652 TI - Observation of frequency-locked coherent terahertz Smith-Purcell radiation. AB - We report the observation of enhanced coherent Smith-Purcell radiation (SPR) at terahertz (THz) frequencies from a train of picosecond bunches of 15 MeV electrons passing above a grating. SPR is more intense than other sources, such as transition radiation, by a factor of Ng, the number of grating periods. For electron bunches that are short compared with the radiation wavelength, coherent emission occurs, enhanced by a factor of Ne, the number of electrons in the bunch. The electron beam consists of a train of Nb bunches, giving an energy density spectrum restricted to harmonics of the 17 GHz bunch train frequency, with an increased energy density at these frequencies by a factor of Nb. We report the first observation of SPR displaying all three of these enhancements, NgNeNb. This powerful SPR THz radiation can be detected with a high signal to noise ratio by a heterodyne receiver. PMID- 15783653 TI - Simulation of heating-compressed fast-ignition cores by petawatt laser-generated electrons. AB - We report on unique particle-in-cell simulations to understand the relativistic electron beam thermalization and subsequent heating of highly compressed plasmas. The simulations yield heated core parameters in good agreement with the GEKKO-PW experimental measurements, given reasonable assumptions of laser-to-electron coupling efficiency and the distribution function of laser-produced electrons. The classical range of the hot electrons exceeds the mass density-core diameter product rhoL by a factor of several. Anomalous stopping appears to be present and is created by the growth and saturation of an electromagnetic filamentation mode that generates a strong back-EMF impeding hot electrons on the injection side of the density maxima. PMID- 15783654 TI - Most electron heat transport is not anomalous; it is a paleoclassical process in toroidal plasmas. AB - It is hypothesized that radial electron heat transport in magnetically confined toroidal plasmas results from paleoclassical Coulomb collision processes (parallel electron heat conduction and magnetic field diffusion). In such plasmas the electron temperature is equilibrated along magnetic field lines a long length L (>> poloidal periodicity length piR0q), which is the minimum of the electron collision length and an effective field line length. Thus, diffusing field lines induce a radial electron heat diffusivity M identical with L/(piR0q) approximately 10>>1 times the magnetic field diffusivity eta/mu0 approximately nue(c/omegap)2. PMID- 15783655 TI - Electron kinetic effects in the nonlinear evolution of a driven ion-acoustic wave. AB - The electron kinetic effects are shown to play an important role in the nonlinear evolution of a driven ion-acoustic wave. The numerical simulation results obtained (i) with a hybrid code, in which the electrons behave as a fluid and the ions are described along the particle-in-cell (PIC) method, are compared with those obtained (ii) with a full-PIC code, in which the kinetic effects on both species are retained. The electron kinetic effects interplay with the usual fluid type nonlinearity to give rise to a broadband spectrum of ion-acoustic waves saturated at a low level, even in the case of a strong excitation. This low asymptotic level might solve the long-standing problem of the small stimulated Brillouin scattering reflectivity observed in laser-plasma interaction experiments. PMID- 15783656 TI - Ultraintense laser-produced fast-electron propagation in gas jets. AB - We study the propagation of fast electrons in a gas at different densities. A large relativistic electron current is produced by focusing a short-pulse ultrahigh-intensity laser on a metallic target. It then propagates in a gas jet placed behind the foil. Shadowgraphy in the gas shows an electron cloud moving at sub-relativistic average velocities. The experiment shows (i) the essential role of the density of background material for allowing propagation of fast electrons, (ii) the importance of the ionization phase which produces free electrons available for the return current, and (iii) the effect of electrostatic fields on fast-electron propagation. PMID- 15783657 TI - Low-level saturation of Brillouin backscattering due to cavity formation in high intensity laser-plasma interaction. AB - Full particle-in-cell simulations of Brillouin backscattering in the high intensity regime are presented. The final state of the strongly nonlinear evolution of the ion-acoustic wave packet consists of its collapse and the formation of a density cavity which is supported by large-amplitude localized electromagnetic fields. The cavitation manifests itself in large oscillations of the reflectivity which is terminated by a low-level saturation with strong kinetic effects. This newly discovered scenario demonstrates the importance of fully kinetic descriptions of Brillouin backscattering for high intensities. PMID- 15783658 TI - Evidence for high-efficiency laser-heated hohlraum performance at 527 nm. AB - A series of experiments conducted on the HELEN laser system [M. J. Norman, Appl. Opt.4120023497], into thermal x-ray generation from hohlraum targets using 527 nm (2omega) wavelength laser light, has shown that it is possible to exceed radiation temperatures previously thought limited by high levels of superthermal or hot electron production or stimulated backscatter. This Letter questions whether the assumptions traditionally applied to hohlraum design with respect to hot plasma filling and the use of 2omega light are too conservative. PMID- 15783659 TI - X-ray intensity fluctuation spectroscopy studies on phase-ordering systems. AB - The order-disorder phase transition in Cu3Au has been studied by x-ray intensity fluctuation spectroscopy. Following a quench from the high-temperature, disordered phase, the ordering kinetics is well described by a universal scaling form that can be measured by time-resolved (incoherent) x-ray scattering. By using coherent scattering, we have measured the fluctuations about this universal scaling form. In the late stages of the ordering process, these fluctuations give a two-time correlation function C(q,t1,t2) which has a scaling form with natural variables deltat=/t1-t2/ and t =(t1+t2) / 2. The scaling form crosses over from linear in t to t1/2. These present the first such results for a nonconserved system. PMID- 15783660 TI - Scaling behavior of the Portevin-Le Chatelier effect in an Al-2.5%Mg alloy. AB - The scaling behavior of the Portevin-Le Chatelier (PLC) effect was studied by deforming an Al-2.5%Mg alloy for a wide range of strain rates. To reveal the exact scaling nature, the time series data of true stress versus time, obtained during deformation, were analyzed by two complementary methods: the finite variance scaling method and the diffusion entropy analysis. From these analyses we could establish that, in the entire span of strain rates, the PLC effect showed the Levy-walk property. PMID- 15783661 TI - Effective temperature in athermal systems sheared at fixed normal load. AB - We perform molecular dynamics simulations of repulsive athermal systems sheared at fixed normal load to study the effective temperature TL defined from time dependent fluctuation-dissipation relations for density. We show that these systems possess two distinct regimes as a function of the ratio TS/V of the granular temperature to the potential energy per particle. At small TS/V, these systems are pressure controlled and TL is set by the normal load. In contrast, they behave as quasiequilibrium systems with TL approximately TS that increases with the shear rate at large TS/V. These results point out several problems with using TL in thermodynamic descriptions of slowly sheared athermal systems. PMID- 15783662 TI - Local spectroscopy and atomic imaging of tunneling current, forces, and dissipation on graphite. AB - Theory predicts that the currents in scanning tunneling microscopy (STM) and the attractive forces measured in atomic force microscopy (AFM) are directly related. Atomic images obtained in an attractive AFM mode should therefore be redundant because they should be similar to STM. Here, we show that while the distance dependence of current and force is similar for graphite, constant-height AFM and STM images differ substantially depending on the distance and bias voltage. We perform spectroscopy of the tunneling current, the frequency shift, and the damping signal at high-symmetry lattice sites of the graphite (0001) surface. The dissipation signal is about twice as sensitive to distance as the frequency shift, explained by the Prandtl-Tomlinson model of atomic friction. PMID- 15783663 TI - Boundary slip on smooth hydrophobic surfaces: intrinsic effects and possible artifacts. AB - We report an accurate determination of the hydrodynamic boundary condition of simple liquids flowing on smooth hydrophobic surfaces using a dynamic surface force apparatus equipped with two independent subnanometer resolution sensors. The boundary slip observed is well defined and does not depend on the scale of investigation from one to several hundreds of nanometers, nor on shear rate up to 5 x 10(3)s(-1). The slip length of 20 nm is in good agreement with theory and numerical simulations concerning smooth nonwetting surfaces. These results disagree with previous data in the literature reporting very high boundary slip on similar systems. We discuss possible origins of large slip length on smooth hydrophobic surfaces due to their contamination by hydrophobic particles. PMID- 15783664 TI - Tip and surface determination from experiments and simulations of scanning tunneling microscopy and spectroscopy. AB - We present a very efficient and accurate method to simulate scanning tunneling microscopy images and spectra from first-principles density functional calculations. The wave functions of the tip and sample are calculated separately on the same footing and propagated far from the surface using the vacuum Green function. This allows us to express the Bardeen matrix elements in terms of convolutions and to obtain the tunneling current at all tip positions and bias voltages in a single calculation. The efficiency of the method opens the door to real time determination of both tip and surface composition and structure, by comparing experiments to simulated images for a variety of precomputed tips. Comparison with the experimental topography and spectra of the Si111-(7 x 7) surface shows a much better agreement with Si than with W tips, implying that the metallic tip is terminated by silicon. PMID- 15783665 TI - Line-on-line coincidence: a new type of epitaxy found in organic-organic heterolayers. AB - We propose a new type of epitaxy, line-on-line coincidence (LOL), which explains the ordering in the organic-organic heterolayer system PTCDA on HBC on graphite. LOL epitaxy is similar to point-on-line coincidence (POL) in the sense that all overlayer molecules lie on parallel, equally spaced lines. The key difference to POL is that these lines are not restricted to primitive lattice lines of the substrate lattice. Potential energy calculations demonstrate that this new type of epitaxy is indeed characterized by a minimum in the overlayer-substrate interaction potential. PMID- 15783666 TI - Determination of the orbital moment and crystal-field splitting in LaTiO3. AB - Utilizing a sum rule in a spin-resolved photoelectron spectroscopic experiment with circularly polarized light, we show that the orbital moment in LaTiO3 is strongly reduced from its ionic value, both below and above the Ne el temperature. Using Ti L2,3 x-ray absorption spectroscopy as a local probe, we found that the crystal-field splitting in the t2g subshell is about 0.12-0.30 eV. This large splitting does not facilitate the formation of an orbital liquid. PMID- 15783667 TI - Quantum critical scaling and the origin of non-fermi-liquid behavior in Sc1 xUxPd3. AB - We used inelastic neutron scattering to study magnetic excitations of Sc1-xUxPd3 for U concentrations (x=0.25, 0.35) near the spin glass quantum critical point (QCP). The excitations are spatially incoherent, broad in energy (E=variant Planck's over 2piomega), and follow omega/T scaling at all wave vectors investigated. Since similar omega/T scaling has been observed for UCu5-xPdx and CeCu6-xAux near the antiferromagnetic QCP, we argue that the observed non-Fermi liquid behavior in these f-electron materials arises from the critical phenomena near a T=0 K phase transition, irrespective of the nature of the transition. PMID- 15783668 TI - Efficient quantum monte carlo energies for molecular dynamics simulations. AB - A method is presented to treat electrons within the many-body quantum Monte Carlo (QMC) approach "on-the-fly" throughout a molecular dynamics (MD) simulation. Our approach leverages the large (10-100) ratio of the QMC electron to MD ion motion to couple the stochastic, imaginary-time electronic and real-time ionic trajectories. This continuous evolution of the QMC electrons results in highly accurate total energies for the full dynamical trajectory at a fraction of the cost of conventional, discrete sampling. We show that this can be achieved efficiently for both ground and excited states with only a modest overhead to an ab initio MD method. The accuracy of this dynamical QMC approach is demonstrated for a variety of systems, phases, and properties, including optical gaps of hot silicon quantum dots, dissociation energy of a single water molecule, and heat of vaporization of liquid water. PMID- 15783670 TI - Ground-state energy of the electron liquid in ultrathin wires. AB - The ground-state energy and the density correlation function of the electron liquid in a thin one-dimensional wire are computed. The calculation is based on an approximate mapping of the problem with a realistic Coulomb interaction law onto exactly solvable models of mathematical physics. This approach becomes asymptotically exact in the limit of a small wire radius but remains numerically accurate even for modestly thin wires. PMID- 15783671 TI - Highly spin-polarized room-temperature tunnel injector for semiconductor spintronics using MgO(100). AB - The spin polarization of current injected into GaAs from a CoFe/MgO(100) tunnel injector is inferred from the electroluminescence polarization from GaAs/AlGaAs quantum well detectors. The polarization reaches 57% at 100 K and 47% at 290 K in a 5 T perpendicular magnetic field. Taking into account the field dependence of the luminescence polarization, the spin injection efficiency is at least 52% at 100 K, and 32% at 290 K. We find a nonmonotonic temperature dependence of the polarization which can be attributed to spin relaxation in the quantum well detectors. PMID- 15783669 TI - Mott-Hubbard transition versus Anderson localization in correlated electron systems with disorder. AB - The phase diagram of correlated, disordered electron systems is calculated within dynamical mean-field theory using the geometrically averaged ("typical") local density of states. Correlated metal, Mott insulator, and Anderson insulator phases, as well as coexistence and crossover regimes, are identified. The Mott and Anderson insulators are found to be continuously connected. PMID- 15783672 TI - Co1-xFexS2: a tunable source of highly spin-polarized electrons. AB - In the emerging field of spin-electronics ideal ferromagnetic electron sources would not only possess a high degree of spin polarization, but would also offer control over the magnitude of this polarization. We demonstrate here that a simple scheme can be utilized to control both the magnitude and the sign of the spin polarization of ferromagnetic CoS2, which we probe with a variety of techniques. The position of the Fermi level is fine-tuned by solid solution alloying with the isostructural diamagnetic semiconductor FeS2, leading to tunable spin polarization of up to 85%. PMID- 15783673 TI - Full current statistics in the regime of weak coulomb interaction. AB - We evaluate the full statistics of the current via a Coulomb island that is strongly coupled to the leads. This strong coupling weakens Coulomb interaction. We show that in this case the effects of the interaction can be incorporated into the renormalization of transmission eigenvalues of the scatterers that connect the island and the leads. We evaluate the Coulomb blockade gap in the current voltage characteristics, the value of the gap being exponentially suppressed as compared to the classical charging energy of the island. PMID- 15783674 TI - Detection of single-electron charging in an individual InAs quantum dot by noncontact atomic-force microscopy. AB - Single-electron charging in an individual InAs quantum dot was observed by electrostatic force measurements with an atomic-force microscope (AFM). The resonant frequency shift and the dissipated energy of an oscillating AFM cantilever were measured as a function of the tip-back electrode voltage, and the resulting spectra show distinct jumps when the tip was positioned above the dot. The observed jumps in the frequency shift, with corresponding peaks in dissipation, are attributed to a single-electron tunneling between the dot and the back electrode governed by the Coulomb blockade effect, and are consistent with a model based on the free energy of the system. The observed phenomenon may be regarded as the "force version" of the Coulomb blockade effect. PMID- 15783675 TI - Lifetime of charge carriers in multiwalled nanotubes. AB - The nature of low-energy excitations in multiwalled nanotubes (MWNTs) is investigated by means of two-color time-resolved photoemission. A careful analysis of the ballistic transport, secondary excitations, and band structure effects was employed in order to extract single electron lifetimes from the observed relaxation trend. It is demonstrated that in the vicinity of the Fermi level the energy dependence of e-e scattering times is inversely proportional to approximately the square of the excitation energy. This result provides strong evidence that electron transport in MWNTs exhibits a Fermi-liquid behavior, indicating that long-range e-e interaction along the tube vanishes due to screening. PMID- 15783676 TI - Observation of the giant stark effect in boron-nitride nanotubes. AB - Bias dependent scanning tunneling microscopy and scanning tunneling spectroscopy have been used to characterize the influence of transverse electric fields on the electronic properties of boron-nitride nanotubes (BNNTs). We find experimental evidence for the theoretically predicted giant Stark effect. The observed giant Stark effect significantly reduces the band gap of BNNTs and thus greatly enhances the utility of BNNTs for nanoscale electronic, electromechanical, and optoelectronic applications. PMID- 15783677 TI - Universal aspects of coulomb-frustrated phase separation. AB - We study the consequences of Coulomb interactions on a system undergoing a putative first order phase transition. In two dimensions (2D), near the critical density, the system is universally unstable to the formation of new intermediate phases, which we call "electronic microemulsion phases," which consist of an intermediate scale mixture of regions of the two competing phases. A corollary is that there can be no direct transition as a function of density from a 2D Wigner crystal to a uniform electron liquid. In 3D, if the strength of the Coulomb interactions exceeds a critical value, no phase separation occurs, while for the weaker Coulomb strength electronic microemulsions are inevitable. This tendency is considerably more pronounced in anisotropic (quasi-2D or quasi-1D) systems, where a devil's staircase of transitions is possible. PMID- 15783678 TI - Photoemission spectroscopic evidence of gap anisotropy in an f-electron superconductor. AB - We used low-temperature ultrahigh-resolution (360 microeV) photoemission spectroscopy with a laser as a photon source (Laser-PES) to study the superconducting (SC) gap of an f-electron superconductor CeRu2. The unique combination of the large escape depth expected from the known universal behavior and extremely high-energy resolution has enabled us to directly measure the bulk SC gap of an f-electron superconductor for the first time. The present study provides direct evidence for an anisotropic SC gap in CeRu2, and also demonstrates the potential of Laser-PES in investigating unconventional superconductivity realized in correlated d- and f-electron superconductors. PMID- 15783679 TI - Giant magnetoresistance in ferromagnet/superconductor superlattices. AB - We show magnetoresistance in excess of 1000% in trilayers containing highly spin polarized La0.7Ca0.3MnO3 and high-Tc superconducting YBa2Cu3O7. This large magnetoresistance is reminiscent of the giant magnetoresistance (GMR) in metallic superlattices but with much larger values, and originates at spin imbalance due to the injection of spin-polarized carriers. Furthermore, in contrast to ordinary GMR, the magnetoresistance is intimately related to the superconductivity in the YBa2Cu3O7 layer and vanishes in the normal state. This result, aside from its fundamental importance, may be of interest for the design of novel spintronic devices based on ferromagnet/superconductor structures. PMID- 15783680 TI - Vortex-rectification effects in films with periodic asymmetric pinning. AB - We study the transport of vortices excited by an ac current in an Al film with an array of nanoengineered asymmetric antidots. The vortex response to the ac current is investigated by detailed measurements of the voltage output as a function of ac current amplitude, magnetic field, and temperature. The measurements revealed pronounced voltage rectification effects which are mainly characterized by the two critical depinning forces of the asymmetric potential. The shape of the net dc voltage as a function of the excitation amplitude indicates that our vortex ratchet behaves in a way very different from standard overdamped models. Rather, the repinning force, necessary to stop vortex motion, is considerably smaller than the depinning force, resembling the behavior of the so-called inertia ratchets. Calculations based on an underdamped ratchet model provide a very good fit to the experimental data. PMID- 15783681 TI - Zener enhancement of quantum tunneling in a two-level superconducting circuit. AB - We have investigated the macroscopic quantum tunneling (MQT) of the phase across a Josephson junction embedded in a superconducting circuit. This system is equivalent to a spin 1/2 particle in a potential energy well. The MQT escape rate of such a particle was recently predicted to be strongly modified when a crossing of its inner Zeeman levels occurs while tunneling. In this regime, we observe a significant enhancement of the MQT rate and compare it to theory. PMID- 15783682 TI - Origin of the anomalous low temperature upturn in the resistivity of the electron doped cuprate superconductors. AB - The temperature, doping, and field dependences of the magnetoresistance (MR) in Pr2-xCexCuO4-delta films are reported. We distinguish between orbital MR, found when the magnetic field is applied perpendicular to the ab planes, and the nearly isotropic spin MR. The latter, the major MR effect in the superconducting samples, appears in the region of the doping-temperature phase diagram where drho/dT<0, or an upturn in the resistivity appears. We conclude that the upturn originates from spin scattering processes. PMID- 15783683 TI - Confined spin waves reveal an assembly of nanosize domains in ferromagneticLa1 xCaxMnO3 (x=0.17,0.2). AB - We report a study of spin waves in ferromagnetic La1-xCaxMnO3, at concentrations x=0.17 and x=0.2 very close to the metallic transition (x=0.225). Below T(C), in the quasimetallic state (T=150 K), nearly q-independent energy levels are observed. They are characteristic of standing spin waves confined into finite size ferromagnetic domains, defined only in the (a,b) plane for x=0.17, and in all directions for x=0.2. They allow an estimation of the domain's size, a few lattice spacings, and of the magnetic coupling constants inside the domains. These constants, anisotropic, are typical of an orbital-ordered state, allowing one to characterize the domains as "hole poor." The precursor state of the collossal magnetoresistance metallic phase appears, therefore, as an assembly of small orbital-ordered domains. PMID- 15783684 TI - Attojoule calorimetry of mesoscopic superconducting loops. AB - We report the first experimental evidence of nontrivial thermal behavior of the simplest mesoscopic system--a superconducting loop. By measuring the specific heat C of an array of 450,000 noninteracting aluminum loops with very high accuracy of approximately 20 fJ/K, we show that the loops go through a periodic sequence of phase transitions (with a period of an integer number of magnetic flux quanta) as the magnetic flux threading each loop is increased. The transitions are well described by the Ginzburg-Landau theory and are accompanied by discontinuities of C of only several thousands of Boltzmann constants kB. PMID- 15783685 TI - Evidence for zero- and pi-phase order parameters of superconducting Nb/Co tri- and pentalayers from the oscillatory behavior of the transition temperature. AB - The oscillation behavior of the superconducting transition temperature Tc as a function of the ferromagnetic Co layer thickness (dCo) has been examined for Nb/Co superconductor(S)/ferromagnetic(F) trilayer series (F/S/F) and pentalayer series (F/S/F/S/F). Tc of the pentalayer series takes a local maximum between dCo=2.0-3.2 nm, where Tc of the trilayer shows a local minimum. This difference in the Tc versus dCo curves provides a clear evidence for the occurrence of the pi phase in the pentalayers, which has been theoretically predicted by Buzdin et al., Radovic et al., and Tagirov. PMID- 15783686 TI - Universality in heavy fermion systems with general degeneracy. AB - We discuss the relation between the T2 coefficient of electrical resistivity A and the T-linear specific-heat coefficient gamma for heavy-fermion systems with general N, where N is the degeneracy of quasiparticles. A set of experimental data reveals that the Kadowaki-Woods relation, A/gamma2=1 x 10(-5) muOmega cm(K mol/mJ)2, collapses remarkably for large-N systems, although this relation has been regarded to be commonly applicable to the Fermi liquids. Instead, based on the Fermi-liquid theory we propose a new relation, A /gamma2=1 x 10(-5) with A =A/1/2N(N-1) and gamma =gamma/1/2N(N-1). This new relation exhibits an excellent agreement with the data for the whole range of degenerate heavy fermions. PMID- 15783687 TI - Spin-wave eigenmodes of permalloy squares with a closure domain structure. AB - Quantized spin-wave eigenmodes in single, 16 nm thick and 0.75 to 4 mum wide square permalloy islands with a fourfold closure domain structure have been investigated by microfocus Brillouin light scattering spectroscopy and time resolved scanning magneto-optical Kerr microscopy. Up to six eigenmodes were detected and classified. The main direction of the spin-wave quantization in the domains was found to be perpendicular to the local static magnetization. An additional less pronounced quantization along the direction parallel to the static magnetization was also observed. PMID- 15783688 TI - Exchange bias and asymmetric reversal in nanostructured dot arrays. AB - The size dependence of exchange bias field HE and coercivity Hc was studied by measuring exchange biased Fe-FeF2 dot arrays in comparison with an unstructured exchange biased Fe-FeF2 bilayer. The domain sizes in the ferromagnet (FM) and the antiferromagnet (AFM) play an important role for exchange bias (EB), and thus interesting phenomena may be expected when the size of an EB system becomes comparable to these sizes. We observe drastic changes of HE and Hc in nanostructured Fe-FeF2, which are unexpected because they appear even at a structure size which is too large for matching with AFM or FM domain size to play a role. We propose that under certain conditions the hysteresis loop is affected differently in the two branches of the reversal by shape anisotropy due to patterning. This is possible because the EB induces a reversal asymmetry already in the unpatterned bilayer system. PMID- 15783689 TI - Giant spin canting in the S=1/2 antiferromagnetic chain [CuPM(NO3)2(H2O)2]n observed by 13C-NMR. AB - We present a combined experimental and theoretical study on copper pyrimidine dinitrate [CuPM(NO3)2(H2O)2]n, a one-dimensional S=1/2 antiferromagnet with alternating local symmetry. From the local susceptibility measured by NMR at the three inequivalent carbon sites in the pyrimidine molecule we deduce a giant spin canting, i.e., an additional staggered magnetization perpendicular to the applied external field at low temperatures. The magnitude of the transverse magnetization, the spin canting of (52+/-4) degrees at 10 K and 9.3 T, and its temperature dependence are in excellent agreement with exact diagonalization calculations. PMID- 15783690 TI - Field-controlled phase separation at the impurity-induced magnetic ordering in the spin-Peierls Magnet CuGeO3. AB - The paramagnetic fraction surviving at the impurity-induced antiferromagnetic phase transition in the spin-Peierls magnet CuGeO3 is found to increase with an external magnetic field. This effect is explained by the competition of the Zeeman interaction and of the exchange interaction of local antiferromagnetic clusters formed on the spin-gap background near impurities. PMID- 15783691 TI - Ferromagnetic Lieb-Mattis theorem. AB - We prove ferromagnetic ordering of energy levels for XXX Heisenberg chains with spins of arbitrary magnitude, thus extending our previous result for the spin-1/2 chain. Ferromagnetic ordering means that the minimum energies in the invariant subspaces of fixed total spin are monotone decreasing as a function of the total spin. This result provides a ferromagnetic analogue of the well-known theorem by Lieb and Mattis about ordering of energy levels in antiferromagnetic and ferrimagnetic systems on bipartite graphs. PMID- 15783692 TI - Far-field optical microscopy with a nanometer-scale resolution based on the in plane image magnification by surface plasmon polaritons. AB - A new far-field optical microscopy capable of reaching nanometer-scale resolution is developed using the in-plane image magnification by surface plasmon polaritons. This approach is based on the optical properties of a metal dielectric interface that may provide extremely large values of the effective refractive index neff up to 10(3) as seen by surface polaritons, and thus the diffraction limited resolution can reach nanometer-scale values of lambda/2neff. The experimental realization of the microscope has demonstrated the optical resolution better than 60 nm at 515 nm illumination wavelength. PMID- 15783693 TI - Direct observation of controlled coupling in an individual quantum dot molecule. AB - We report the direct observation of quantum coupling in individual quantum dot molecules and its manipulation using static electric fields. A pronounced anticrossing of different excitonic transitions is observed as the electric field is tuned. A comparison of our experimental results with theory shows that the observed anticrossing occurs between excitons with predominant spatially direct and indirect character and reveals a field driven transition of the nature of the molecular ground state exciton wave function. Finally, the interdot quantum coupling strength is deduced optically and its dependence on the interdot separation is calculated. PMID- 15783694 TI - Quantum fluctuations of the refractive index near the interface between a metal and a nonlinear dielectric. AB - Zero-point fluctuations of surface plasmon modes near the interface between a metal and a nonlinear dielectric are shown to produce a thin layer of shifted fluctuating dielectric constant near the interface. The shift of the dielectric constant in this layer may be sufficiently large to produce multiple metastable states of the surface plasmon vacuum. PMID- 15783695 TI - Decoherence of rabi oscillations in a single quantum dot. AB - We develop a realistic model of Rabi oscillations in a quantum-dot photodiode. Based in a multiexciton density matrix formulation we show that for short pulses the two-level model fails and higher levels should be taken into account. This affects some of the experimental conclusions, such as the inferred efficiency of the state rotation (population inversion) and the deduced value of the dipole interaction. We also show that the damping observed cannot be explained using constant rates with fixed pulse duration. We demonstrate that the damping observed is in fact induced by an off-resonant excitation to or from the continuum of wetting layer states. Our model describes the nonlinear decoherence behavior observed in recent experiments. PMID- 15783696 TI - Dynamics of water confined on a nanometer length scale in reverse micelles: ultrafast infrared vibrational echo spectroscopy. AB - The dynamics of water, confined on a nanometer length scale (1.7 to 4.0 nm) in sodium bis-(2-ethylhexyl) sulfosuccinate reverse micelles, is directly investigated using frequency resolved infrared vibrational echo experiments. The data are compared to bulk water and salt solution data. The experimentally determined frequency-frequency correlation functions show that the confined water dynamics is substantially slower than bulk water dynamics and is size dependent. The fastest dynamics (approximately 50 fs) is more similar to bulk water, while the slowest time scale dynamics is much slower than water, and, in analogy to bulk water, reflects the making and breaking of hydrogen bonds. PMID- 15783697 TI - Phase evolution of solitonlike optical pulses during excitonic Rabi flopping in a semiconductor. AB - We demonstrate that the temporal pulse phase remains essentially unaltered before separate phase characteristics are developed when propagating high-intensity pulses coherently on the exciton resonance of an optically thick semiconductor. This behavior is a clear manifestation of self-induced transmission and pulse breakup into soliton-like pulses due to Rabi flopping of the carrier density. Experiments using a novel fast-scan cross-correlation frequency-resolved optical gating (XFROG) method are in good agreement with numerical calculations based on the semiconductor Bloch equations. PMID- 15783698 TI - Bonding in liquid carbon studied by time-resolved x-ray absorption spectroscopy. AB - Even the most basic properties of liquid carbon have long been debated due to the challenge of studying the material at the required high temperature and pressure. Liquid carbon is volatile and thus inherently transient in an unconstrained environment. In this paper we use a new technique of picosecond time-resolved x ray absorption spectroscopy to study the bonding of liquid carbon at densities near that of the solid. As the density of the liquid increases, we see a change from predominantly sp-bonded atomic sites to a mixture of sp, sp2, and sp3 sites and compare these observations with molecular dynamics simulations. PMID- 15783699 TI - Dispersive terahertz gain of a nonclassical oscillator: BLOCH oscillation in semiconductor superlattices. AB - We have directly determined the spectral shape of the complex conductivities of Bloch oscillating electrons by using the time-domain terahertz (THz) electro optic sampling technique, and presented experimental evidence for a dispersive Bloch gain in superlattices. This unique dispersive gain without population inversion arises from a nonclassical nature of Bloch oscillations; that is, the phase of the Bloch oscillation is shifted by pi/2 from that of the semiclassical charged harmonic oscillation when driven by the same ac field. By increasing the bias electric field, the gain bandwidth reached approximately 3 THz in our particular sample. PMID- 15783700 TI - Nematic phase transitions in mixtures of thin and thick colloidal rods. AB - We report experimental measurements of the phase behavior of mixtures of thin (charged semiflexible fd virus) and thick (fd-PEG, fd virus covalently coated with polyethylene glycol) rods with diameter ratio varying from 3.7 to 1.1. The phase diagrams of the rod mixtures reveal isotropic-nematic, isotropic-nematic nematic, and nematic-nematic coexisting phases with increasing concentration. In stark contrast to predictions from earlier theoretical work, we observe a nematic nematic coexistence region bound by a lower critical point. Moreover, we show that a rescaled Onsager-type theory for binary hard-rod mixtures qualitatively describes the observed phase behavior. PMID- 15783701 TI - Polar anchoring strength of a tilted nematic: confirmation of the dual easy axis model. AB - The polar anchoring strength coefficient W and polar pretilt angle theta0 were measured simultaneously for the liquid crystal pentylcyanobiphenyl at a rubbed polyimide alignment layer that is ordinarily used for vertical alignment. It was found that W proportional theta(2)0 over the range 0 degrees < or =theta0 less or similar to 35 degrees . The results provide a confirmation of the dual easy axis model, wherein the liquid crystal director adopts an equilibrium orientation theta0 at the substrate that is determined by competition between a pair of preferred orientation directions. PMID- 15783702 TI - Critical branching captures activity in living neural networks and maximizes the number of metastable States. AB - Recent experimental work has shown that activity in living neural networks can propagate as a critical branching process that revisits many metastable states. Neural network theory suggests that attracting states could store information, but little is known about how a branching process could form such states. Here we use a branching process to model actual data and to explore metastable states in the network. When we tune the branching parameter to the critical point, we find that metastable states are most numerous and that network dynamics are not attracting, but neutral. PMID- 15783703 TI - Magic number 7+/-2 in networks of threshold dynamics. AB - Information processing by random feed-forward networks consisting of units with a sigmoidal input-output response is studied by focusing on the dependence of its outputs on the number of parallel paths M. It is found that the system leads to a combination of on-off outputs when M less or similar to 7, while for M greater or similar to 7, chaotic dynamics arises, resulting in a continuous distribution of outputs. This universality of the critical number M approximately 7 is explained by a combinatorial explosion, i.e., the dominance of factorial over exponential increase. The relevance of the result to biological problems is briefly discussed. PMID- 15783704 TI - Nanoparticle-free single molecule anti-stokes Raman spectroscopy. AB - In the absence of large, plasmon-supporting nanoparticles, biocompatible dendrimer- and peptide-encapsulated few-atom Ag nanoclusters produce scaffold specific single molecule (SM) Stokes and anti-Stokes Raman scattering. The strong SM vibrational signatures are enhanced by the Agn transitions in nanoparticle free samples and cannot arise from plasmon enhancement. Characteristic SM-Raman intermittency is observed, with antibunching of the underlying Agn emission directly confirming the SM nature of the emissive species. PMID- 15783706 TI - Simulation study of spatiotemporal correlations of earthquakes as a stick-slip frictional instability. AB - Spatiotemporal correlations of earthquakes are studied numerically on the basis of the one-dimensional spring-block (Burridge-Knopoff) model. As large events approach, the frequency of smaller events gradually increases, while, just before the mainshock, it is dramatically suppressed in a close vicinity of the epicenter of the upcoming mainshock, a phenomenon closely resembling the "Mogi doughnut." PMID- 15783705 TI - Colloidal interactions and self-assembly using DNA hybridization. AB - The specific binding of complementary DNA strands has been suggested as an ideal method for directing the controlled self-assembly of microscopic objects. We report the first direct measurements of such DNA-induced interactions between colloidal microspheres, as well as the first colloidal crystals assembled using them. The interactions measured with our optical tweezer method can be modeled in detail by well-known statistical physics and chemistry, boding well for their application to directed self-assembly. The microspheres' binding dynamics, however, have a surprising power-law scaling that can significantly slow annealing and crystallization. PMID- 15783707 TI - Effect of congestion costs on shortest paths through complex networks. AB - We analyze analytically the effect of congestion costs within a physically relevant, yet exactly solvable, network model featuring central hubs. These costs lead to a competition between centralized and decentralized transport pathways. In stark contrast to conventional no-cost networks, there now exists an optimal number of connections to the central hub in order to minimize the shortest path. Our results shed light on an open problem in biology, informatics, and sociology, concerning the extent to which decentralized versus centralized design benefits real-world complex networks. PMID- 15783708 TI - Density profiles of strongly interacting trapped fermi gases. AB - We study density profiles in trapped fermionic gases, near Feshbach resonances, at all T< or =Tc and in the near Bose-Einstein condensation and unitary regimes. For the latter, we characterize and quantify the generally neglected contribution from noncondensed Cooper pairs. As a consequence of these pairs, our profiles are rather well fit to a Thomas-Fermi (TF) functional form, and equally well fit to experimental data. Our work lends support to the notion that TF fits can be used in an experimental context to obtain information about the temperature. PMID- 15783709 TI - Exact results for one-dimensional disordered bosons with strong repulsion. AB - We study one-dimensional disordered bosons with strong repulsive interactions. A Bose-Fermi mapping expresses this problem in terms of noninteracting Anderson localized fermions, whereby known results for the distribution function of the local density of states, the spectral statistics, and density-density correlations can be transferred to this new domain of applicability. We show that disorder destroys bosonic quasi-long-range order by calculating the momentum distribution, and comment on the experimental observability of these predictions in ultracold atomic gases. PMID- 15783710 TI - Feshbach-resonant interactions in 40K and 6Li degenerate Fermi gases. AB - We theoretically examine a system of Fermi degenerate atoms coupled to bosonic molecules by a Feshbach resonance, focusing on the superfluid transition to a molecular Bose-Einstein condensate dressed by Cooper pairs of atoms. This problem raises interest because it is unclear at present whether bimodal density distributions observed recently in 40K and 6Li are due to a condensate of bosonic molecules or fermionic atom pairs. As opposed to 40K, we find that any measurable fraction of above-threshold bosonic molecules is necessarily absent for the 6Li system in question, which strongly implicates Cooper pairs as the culprit behind its bimodal distributions. PMID- 15783711 TI - Direct imaging of spatially modulated superfluid phases in atomic fermion systems. AB - It is proposed that the spatially modulated superfluid phase, or the Fulde Ferrell-Larkin-Ovchinnikov state could be observed in resonant fermion atomic condensates which are realized recently. We examine optimal experimental setups to achieve it by solving the Bogoliubov-de Gennes equation for both idealized one dimensional and realistic three-dimensional cases. The spontaneous modulation of this superfluid is shown to be directly imaged as the density profiles either by optical absorption or by Stern-Gerlach experiments. PMID- 15783712 TI - Detecting genuine multipartite entanglement with two local measurements. AB - We present entanglement witness operators for detecting genuine multipartite entanglement. These witnesses are robust against noise and require only two local measurement settings when used in an experiment, independent of the number of qubits. This allows detection of entanglement for an increasing number of parties without a corresponding increase in effort. The witnesses presented detect states close to Greenberger-Horne-Zeilinger, cluster, and graph states. Connections to Bell inequalities are also discussed. PMID- 15783713 TI - Random decoupling schemes for quantum dynamical control and error suppression. AB - We present a general control-theoretic framework for constructing and analyzing random decoupling schemes, applicable to quantum dynamical control of arbitrary finite-dimensional composite systems. The basic idea is to design the control propagator according to a random rather than deterministic path on a group. We characterize the performance of random decoupling protocols, and identify control scenarios where they can significantly weaken time scale requirements as compared to cyclic counterparts. Implications for reliable quantum computation are discussed. PMID- 15783714 TI - Entropy, entanglement, and area: analytical results for harmonic lattice systems. AB - We revisit the question of the relation between entanglement, entropy, and area for harmonic lattice Hamiltonians corresponding to discrete versions of real free Klein-Gordon fields. For the ground state of the d-dimensional cubic harmonic lattice we establish a strict relationship between the surface area of a distinguished hypercube and the degree of entanglement between the hypercube and the rest of the lattice analytically, without resorting to numerical means. We outline extensions of these results to longer ranged interactions, finite temperatures, and for classical correlations in classical harmonic lattice systems. These findings further suggest that the tools of quantum information science may help in establishing results in quantum field theory that were previously less accessible. PMID- 15783715 TI - Nonlinear amplification of small spin precession using long-range dipolar interactions. AB - In measurements of small signals using spin precession the precession angle usually grows linearly in time. We show that a dynamic instability caused by spin interactions can lead to an exponentially growing spin-precession angle, amplifying small signals and raising them above the noise level of a detection system. We demonstrate amplification by a factor of greater than 8 of a spin precession signal due to a small magnetic field gradient in a spherical cell filled with hyperpolarized liquid 129Xe. This technique can improve the sensitivity in many measurements that are limited by the noise of the detection system, rather than the fundamental spin-projection noise. PMID- 15783716 TI - Cutoff anti-de Sitter space/conformal-field-theory duality and the quest for braneworld black holes. AB - Significant evidence is presented in favor of the holographic conjecture that "4D black holes localized on the brane found by solving the classical bulk equations in AdS5 are quantum corrected black holes and not classical ones." The quantum correction to the Newtonian potential is computed using a numerical computation of in Schwarzschild spacetime for matter fields in the zero-temperature Boulware vacuum state. For the conformally invariant scalar field the leading order term is equivalent to that previously obtained in the weak-field approximation using Feynman diagrams and which has been shown to be equivalent, via the anti-de Sitter space/conformal-field-theory (AdS/CFT) duality, to the analogous calculation in Randall-Sundrum braneworlds. The 4D backreaction equations are used to make a prediction about the existence and the possible spacetime structure of macroscopic static braneworld black holes. PMID- 15783717 TI - Hawking radiation in sonic black holes. AB - I present a microscopic description of Hawking radiation in sonic black holes. A one-dimensional Fermi-degenerate liquid squeezed by a smooth barrier forms a transonic flow, a sonic analog of a black hole. The quantum treatment of the noninteracting case establishes a close relationship between sonic Hawking radiation and quantum tunneling through the barrier. Quasiparticle excitations appear at the barrier and are then radiated with a thermal distribution in exact agreement with Hawking's formula. The signature of the radiation can be found in the dynamic structure factor, which can be measured in a scattering experiment. The possibility for experimental verification of this new transport phenomenon for ultracold atoms is discussed. PMID- 15783718 TI - Inverse-scattering theory and the density perturbations from inflation. AB - We show how to use inverse-scattering theory as the basis for the inflationary reconstruction program, the goal of which is to gain information about the physics which drives inflation. Inverse-scattering theory provides an effective and well-motivated procedure, having a sound mathematical basis and being of sufficient generality that it can be considered the foundation for a nonparametric reconstruction program. We show how simple properties of the power spectrum translate directly into statements about the evolution of the background geometry during inflation. PMID- 15783719 TI - Failure of mean field theory at large N. AB - We study strongly coupled lattice QCD with N colors of staggered fermions in 3+1 dimensions. While mean field theory describes the low temperature behavior of this theory at large N, it fails in the scaling region close to the finite temperature second order chiral phase transition. The universal critical region close to the phase transition belongs to the 3D XY universality class even when N becomes large. This is in contrast to Gross-Neveu models where the critical region shrinks as N (the number of flavors) increases and mean field theory is expected to describe the phase transition exactly in the limit of infinite N. Our work demonstrates that infrared fluctuations can be important close to second order phase transitions even when N is strictly infinite. PMID- 15783720 TI - Electroweak baryogenesis in a supersymmetric U(1)' model. AB - We construct an anomaly-free supersymmetric U(1)' model with a secluded U(1)' breaking sector. We study the one-loop effective potential at finite temperature and show that there exists a strong enough first order electroweak phase transition for electroweak baryogenesis (EWBG) because of the large trilinear term AhhSHdHu in the tree-level Higgs potential. Unlike in the minimal supersymmetric standard model, the lightest top squark can be very heavy. We consider the nonlocal EWBG mechanism in the thin wall regime and find that within uncertainties the observed baryon number can be generated from the tau lepton contribution, with the secluded sector playing an essential role. The chargino and neutralino contributions and the implications for the Z' mass and electric dipole moments are briefly discussed. PMID- 15783722 TI - Determining the sign of the b-->sgamma amplitude. AB - The latest Belle and BABAR measurements of the inclusive B -->Xsl+l- branching ratio have smaller errors and lower central values than the previous ones. We point out that these results indicate that the sign of the b-->sgamma amplitude is the same as in the standard model. This underscores the importance of B - >Xsl+l- in searches for new physics, and may be relevant for neutralino-dark matter analyses within the minimal supersymmetric standard model. PMID- 15783723 TI - Pentaquarks uudds with one color sextet diquark. AB - The masses of pentaquarks uudds are calculated within the framework of a semirelativistic effective QCD Hamiltonian using a diquark picture. This approximation allows a correct treatment of the confinement, assumed here to be similar to a Y junction. With only color antitriplet diquarks, the mass of the pentaquark candidate Theta with positive parity is found around 2.2 GeV. It is shown that, if a color sextet diquark is present, the lowest uudds pentaquark is characterized by a much smaller mass with a negative parity. A mass below 1.7 GeV is computed if the masses of the color antitriplet and color sextet diquarks are taken similar. PMID- 15783725 TI - Discovery of dumbbell-shaped Cs*Hen exciplexes in solid 4He. AB - We have observed several new spectral features in the fluorescence of cesium atoms implanted in the hcp phase of solid helium following laser excitation to the 62P states. Based on calculations of the emission spectra using semiempirical Cs-He pair potentials the newly discovered lines can be assigned to the decay of specific Cs*Hen exciplexes: an apple-shaped Cs(APi3/2)He2 and a dumbbell-shaped Cs(APi1/2)Hen exciplex with a well-defined number n of bound helium atoms. While the former has been observed in other environments, it was commonly believed that exciplexes with n>2 might not exist. The calculations suggest Cs(APi1/2)He7 to be the most probable candidate for that exciplex, in which the helium atoms are arranged on a ring around the waist of the dumbbell-shaped electronic density distribution of the cesium atom. PMID- 15783726 TI - Lithium ionization by a strong laser field. AB - We study ab initio computations of the interaction of lithium with a strong laser field. Numerical solutions of the time-dependent fully correlated three-particle Schrodinger equation restricted to the one-dimensional soft-core approximation are presented. Our results show a clear transition from nonsequential to sequential double ionization for increasing intensities. Nonsequential double ionization is found to be sensitive to the spin configuration of the ionized pair. This asymmetry, also found in experiments of photoionization of Li with synchrotron radiation, shows evidence of the influence of the exclusion principle on the underlying rescattering mechanism. PMID- 15783721 TI - Observation of B0-->D*sJ(2317)+K- decay. AB - The decays B0-->D+sJK- and B0-->D-sJpi+ are studied for the first time. A significant signal is observed in the B0-->D*sJ(2317)+K- decay channel with B(B0- >D*sJ(2317)+K-) x B(D*sJ(2317)+-->D+spi0)=(5.3(+1.5)(-1.3)+/-0.7+/-1.4) x 10(-5). No signals are observed in the B0-->D*sJ(2317)-pi+, B0-->DsJ(2460)+K-, and B 0- >DsJ(2460)-pi+ decay modes, and upper limits are obtained. The analysis is based on a data set of 140 fb(-1) collected by the Belle experiment at the asymmetric e+e- collider KEKB. PMID- 15783727 TI - Theory of electron attachment to CO2 clusters. AB - A theory of electron collisions with van der Waals clusters is developed which is capable of describing the vibrational Feshbach resonances (VFRs) recently observed in electron attachment to CO2 clusters. The calculations also predict vibrational excitation (VE) of one molecular unit in a cluster environment and demonstrate a strong influence of the VFRs on the VE cross sections. PMID- 15783728 TI - Observation of cold, long-lived antiprotonic helium ions. AB - Cold, two-body antiprotonic helium ions p 4He2+ and p 3He2+ with 100-ns-scale lifetimes, occupying circular states with the quantum numbers ni=28-32 and li=ni 1 have been observed. They were produced by cooling three-body antiprotonic helium atoms in an ultra-low-density helium target at temperature T approximately 10 K by atomic collisions, and then removing their electrons by inducing a laser transition to an autoionizing state. The lifetimes of p 3He2+ against annihilation induced by collisions were shorter than those of p 4He2+, and decreased for larger-ni states. PMID- 15783729 TI - Two-photon imaging with thermal light. AB - We report the first experimental demonstration of two-photon imaging with a pseudothermal source. Similarly to the case of entangled states, a two-photon Gaussian thin lens equation is observed, indicating EPR type correlation in position. We introduce the concepts of two-photon coherent and two-photon incoherent imaging. The differences between the entangled and the thermal cases are explained in terms of these concepts. PMID- 15783730 TI - Terahertz response of a microfabricated rod-split-ring-resonator electromagnetic metamaterial. AB - The first electromagnetic metamaterials (EM3) produced by microfabrication are reported. They are based on the rod-split-ring-resonator design as proposed by Pendry et al. [IEEE Trans. Microwave Theory Tech. 47, 2075 (1999)] and experimentally confirmed by Smith et al. [Phys. Rev. Lett. 84, 4184 (2000)] in the GHz frequency range. Numerical simulation and experimental results from far infrared (FIR) transmission spectroscopy support the conclusion that the microfabricated composite material is EM3 in the range 1-2.7 THz. This extends the frequency range in which EM3 are available by about 3 orders of magnitude well into the FIR, thereby widely opening up opportunities to verify the unusual physical implications on electromagnetic theory as well as to build novel electromagnetic and optical devices. PMID- 15783731 TI - Nonlinear optics with stationary pulses of light. AB - We show that the recently demonstrated technique for generating stationary pulses of light [M. Bajcsy, A. S. Zibrov, and M. D. Lukin, Nature (London) 426, 638 (2003)] can be extended to localize optical pulses in all three spatial dimensions in a resonant atomic medium. This method can be used to dramatically enhance the nonlinear interaction between weak optical pulses. In particular, we show that an efficient Kerr-like interaction between two pulses can be implemented as a sequence of several purely linear optical processes. The resulting process may enable coherent interactions between single photon pulses. PMID- 15783732 TI - Color-tunable organic microcavity laser array using distributed feedback. AB - Distributed feedback microstructures play a fundamental role in confining and manipulating light to obtain lasing in media with gain. Here, we present an innovative array of organic, color-tunable microlasers which are intrinsically phase locked. Dye-doped helixed liquid crystals were embedded within periodic, polymeric microchannels sculptured by light through a single-step process. The helical superstructure was oriented along the microchannels; the lasing was observed along the same direction at the red edge of the stop band. Several physical and technological advantages arise from this engineered heterostructure: a high quality factor of the cavity, ultralow lasing threshold, and thermal and electric control of the lasing wavelength and emission intensity. This level of integration of guest-host systems, embedded in artificially patterned small sized structures, might lead to new photonic chip architectures. PMID- 15783733 TI - Coherence controlled soliton interactions. AB - We demonstrate theoretically and subsequently observe in experiment a novel type of soliton interaction when a pair of closely spaced spatial optical solitons as a whole is made partially incoherent. We explain how the character of the soliton interaction can be controlled by the total partial incoherence, and show a possibility to change the soliton interaction from attractive to repulsive, or vice versa, near a certain threshold in the coherence parameter. PMID- 15783734 TI - Excitability mediated by localized structures in a dissipative nonlinear optical cavity. AB - We find and characterize an excitability regime mediated by localized structures in a dissipative nonlinear optical cavity. The scenario is that stable localized structures exhibit a Hopf bifurcation to self-pulsating behavior, that is followed by the destruction of the oscillation in a saddle-loop bifurcation. Beyond this point there is a regime of excitable localized structures under the application of suitable perturbations. Excitability emerges from the spatial dependence since the system does not exhibit any excitable behavior locally. We show that the whole scenario is organized by a Takens-Bogdanov codimension-2 bifurcation point. PMID- 15783735 TI - Scaling behavior for the onset of convection in a colloidal suspension. AB - We investigate the early stages of mass convection in a colloidal suspension at high solutal Rayleigh number Ras. From the time evolution of shadowgraph images and by assuming a diffusive growth of the boundary layers we obtain an indirect measurement of the concentration boundary layer thickness delta* at the onset of convection. We show that the dimensionless boundary layer thickness delta=delta*/d scales as Ra-ps, where Ras=Rasdelta is a modified solutal Rayleigh number for convection which accounts for the actual density unbalance and d is the thickness of the sample layer. This scaling behavior is analogous to that reported at steady state for turbulent convection in simple fluids. We find p=0.35, a value compatible with the exponent 1/3, reported for turbulent heat convection in simple fluids at steady state. PMID- 15783724 TI - Open charm yields in d+Au collisions at squareroot[sNN]=200 GeV. AB - Midrapidity open charm spectra from direct reconstruction of D0(D0)-->K-/+pi+/- in d+Au collisions and indirect electron-positron measurements via charm semileptonic decays in p+p and d+Au collisions at squareroot[sNN]=200 GeV are reported. The D0(D0) spectrum covers a transverse momentum (pT) range of 0.12 A). We report on the first such transition in a quaternary system based on a transition element. Metallic LaMg2Ni absorbs hydrogen near ambient conditions, forming the nonmetallic hydride LaMg2NiH7 which has a nearly unchanged metal host structure (atom shifts <0.7 A). The transition is induced by a charge transfer of conduction electrons into tetrahedral [NiH4]4- complexes having closed-shell electron configurations. PMID- 15783760 TI - Pumped spin-current and shot-noise spectra of a single quantum dot. AB - We exploit the pumped spin-current and current noise spectra under equilibrium conditions in a single quantum dot connected to two normal leads as an electrical scheme for detection of the electron spin resonance (ESR) and decoherence. We propose spin-resolved quantum rate equations with correlation functions in Laplace space for the analytical derivation of the zero-frequency auto- and cross shot noise spectra of charge and spin current. Our results show that in the strong Coulomb blockade regime, ESR-induced spin flip generates a finite spin current and quantum partition noises in the absence of net charge transport. Moreover, spin shot noise is closely related to the magnetic Rabi frequency and decoherence and would be a sensitive tool to measure them. PMID- 15783761 TI - Intrinsic spin and orbital angular momentum Hall effect. AB - A generalized definition of intrinsic and extrinsic transport coefficients is introduced. We show that transport coefficients from the intrinsic origin are solely determined by local electronic structure, and thus the intrinsic spin Hall effect is not a transport phenomenon. The intrinsic spin Hall current is always accompanied by an equal but opposite intrinsic orbital angular momentum Hall current. We prove that the intrinsic spin Hall effect does not induce a spin accumulation at the edge of the sample or near the interface. PMID- 15783762 TI - Electron delocalization and dimerization in solid C59N doped C60 fullerene. AB - Electron spin resonance and ab initio electronic structure calculations show an intricate relation between molecular rotation and chemical bonding in the dilute solid solution. The unpaired electron of C59N is delocalized over several C60 molecules above 700 K, while at lower temperatures it remains localized within short range. The data suggest that below 350 K rigid C59N-C60 heterodimers are formed in thermodynamic equilibrium with dissociated rotating molecules. The structural fluctuations between heterodimers and dissociated molecules are accompanied by simultaneous electron spin transfer between C60 and C59N molecules. The calculation confirms that in the C59N-C60 heterodimer the spin density resides mostly on the C60 moiety, while it is almost entirely on C59N in the dissociated case. PMID- 15783763 TI - Gate-dependent magnetoresistance phenomena in carbon nanotubes. AB - We report on the first experimental study of the magnetoresistance of double walled carbon nanotubes under a magnetic field as large as 50 T. By varying the field orientation with respect to the tube axis, or by gate-mediated shifting the Fermi level position, evidence for unconventional magnetoresistance is presented and interpreted by means of theoretical calculations. PMID- 15783764 TI - Phonon-induced resistivity of electron liquids in quantum wires. AB - We study the resistivity of a quantum wire caused by backscattering of electrons by acoustic phonons. In the presence of Coulomb interactions, backscattering is strongly enhanced at low temperatures due to Luttinger liquid effects. Information about the strength of the interactions can be obtained from a measurement of the temperature dependence of the resistivity. PMID- 15783765 TI - Spontaneous relaxation of a charge qubit under electrical measurement. AB - In this work we first derive a generalized conditional master equation for quantum measurement by a mesoscopic detector, then study the readout characteristics of qubit measurement where a number of remarkable new features are found. The work would, in particular, highlight the qubit spontaneous relaxation effect induced by the measurement itself rather than an external thermal bath. PMID- 15783766 TI - Tilt of pancake vortex stacks in layered superconductors in the crossing lattice regime. AB - We study crossing vortices in strongly anisotropic Bi2Sr2CaCu2O8+delta single crystals. Using scanning Hall probe microscopy and Bitter decoration techniques, we find an asymmetry of magnetic field profiles produced by pancake vortices (PVs), which are interacting with Josephson vortices (JVs), near the surface of the crystal. We attribute the observed asymmetry to a substantial tilt (14-18 degrees) of PV stacks, which is produced by the torque due to the surface currents and JVs. We calculate the tilt angle and obtain agreement with experimental data when the irreversible in-plane magnetization is included. A further refinement to the model is considered which accounts for a reduction in the PV stack line tension near the sample surface. PMID- 15783767 TI - Frequency-dependent thermal response of the charge system and the restricted sum rules of La2-xSrxCuO4. AB - In La2-xSrxCuO4 (LSCO) the spectral weight W=integralOmega0sigma(ab)1(omega,T)domega [where sigma(ab)1(omega,T) is the ab plane conductivity] obeys the same law W=W0-BOmegaT2 as in a conventional metal such as gold, for any Omega up to the plasma edge. However, in LSCO BOmega points toward correlation effects and, unlike in gold, is related to an energy scale tT< K+ pi(-) decays and measurement of the doubly Cabibbo-suppressed decay rate. AB - We have searched for mixing in the D(0)-D (0) system by measuring the decay-time distribution of D(0) --> K(+) pi(-) decays. The analysis uses 90 fb(-1) of data collected by the Belle detector at the KEKB e(+) e(-) collider. We fit the decay time distribution for the mixing parameters x' and y' and also for the parameter R(D), which is the ratio of the rate for the doubly-Cabibbo-suppressed decay D(0) -> K+ pi(-) to that for the Cabibbo-favored decay D(0)--> K-pi(+). We do these fits both assuming CP conservation and allowing for CP violation. We use a frequentist method to obtain a 95% C.L. region in the x'(2) - y' plane. Assuming no mixing, we measure R(D) = (0.381 +/- 0.017(+0.008)(-0.016))%. PMID- 15783805 TI - Traces of the theta+ pentaquark in K+ -nucleus dynamics. AB - Long-standing anomalies in K(+)-nucleus integral cross sections could be resolved by extending the impulse-approximation trho optical-potential framework to incorporate K(+) absorption on pairs of nucleons. Substantially improved fits to the data at p(lab) approximately 500-700 MeV/c are obtained. An upper bound on the absorption cross section per nucleon is derived, sigmaabs((K(+))/A approximately 3.5 mb. We conjecture that the underlying microscopic absorption process is K(+) nN--> Theta+ N, where Theta(+) (1540) is the newly discovered exotic Y = 2, I = 0, Z = 1 pentaquark baryon, and estimate that sigma(K(+) d--> Theta(+) p) is a fraction of millibarn. Comments are made on Theta(+) production reactions on nuclei. PMID- 15783806 TI - Apparent thermalization due to plasma instabilities in the quark-gluon plasma. AB - Hydrodynamical modeling of heavy-ion collisions at RHIC suggests that the quark gluon plasma (QGP) "thermalizes" in a remarkably short time scale, about 0.6 fm/c. We argue that this should be viewed as indicating fast isotropization, but not necessarily complete thermalization, of the nonequilibrium QGP. Non-Abelian plasma instabilities can drive local isotropization of an anisotropic QGP on a time scale which is faster than ordinary perturbative scattering processes. As a result, we argue that theoretical expectations based on weak-coupling analysis are not necessarily in conflict with hydrodynamic modeling of the early part of RHIC collisions, provided one recognizes the key role of non-Abelian plasma instabilities. PMID- 15783807 TI - Polarization transfer in 4He(e-->,e'p-->)3H: is the ratio G E p/G M p ) modified in the nuclear medium? AB - Polarization observables in the (4)He(e-->,e'p-->)(3)H reaction are calculated using accurate three- and four-nucleon bound-state wave functions, a realistic model for the nuclear electromagnetic current operator, and a treatment of final state interactions with an optical potential. In contrast to earlier studies, no significant discrepancies are found between theory and experiment both for the ratio of transverse to longitudinal polarization transfers and for the induced polarization, when free-nucleon electromagnetic form factors are used in the current operator. The present results challenge the current interpretation of the experimental data in terms of medium-modified form factors. PMID- 15783808 TI - Measurement of the analyzing power in p-->d-->(p p)n with a fast forward 1S0 proton pair. AB - A measurement of the analyzing power A(y) of the p-->d--> (p p) + n reaction was carried out at the ANKE spectrometer at COSY at beam energies of 0.5 and 0.8 GeV by detection of a fast forward proton pair of small excitation energy E(pp) < 3 MeV. The S-wave dominance in the fast diproton is experimentally demonstrated in this reaction. While at T(p) = 0.8 GeV the measured analyzing power almost vanishes, it rises to nearly unity at T(p) = 0.5 GeV for neutrons emitted at theta(c.m.)(n) = 167 degrees. The results are compared with a model taking into account one-nucleon exchange, single scattering, and Delta(1232) excitation in the intermediate state. The model describes fairly well the unpolarized cross section obtained earlier and the analyzing power at 0.8 GeV; it fails to reproduce A(y) at 0.5 GeV. PMID- 15783809 TI - Transport coefficients of a gluon plasma. AB - Transport coefficients of gluon plasma are calculated for an SU(3) pure gauge model by lattice QCD simulations on 16(3) x 8 and 24(3) x 8 lattices. Simulations are carried out at slightly above the deconfinement transition temperature T(c), where a new state of matter is currently being pursued in BNL RHIC experiments. Our results show that the ratio of the shear viscosity to the entropy is less than one and the bulk viscosity is consistent with zero in the region 1.4 < or = T/T(c) < or = 1.8. PMID- 15783810 TI - Relativistic continuum-continuum coupling in the dissociation of halo nuclei. AB - A relativistic coupled-channels theory for the calculation of dissociation cross sections of halo nuclei is developed. A comparison with nonrelativistic models is done for the dissociation of (8)B projectiles. It is shown that neglecting relativistic effects leads to sizable inaccuracies in the extraction of the astrophysical S factor for the proton + beryllium radiative capture reaction. PMID- 15783811 TI - Experimental determination of the nu5 cis-bending vibrational frequency and Renner-Teller structure in ground state (X2Piu) C2H2+ using laser induced reactions. AB - The spectrum of the nu(5) cis-bending vibration of ground state (X(2)Pi(u)) C(2)H(2)(+) has been recorded applying the method of laser induced reactions in a low-temperature 22-pole ion trap. It is obtained by counting the number of products of the reaction C(2)H(2)(+) (v(5) = 1) + H(2)--> C(2)H(3)(+) + H as a function of the laser wavelength. The vibronic transitions Delta-Pi and Sigma-Pi with their corresponding spin-orbit and Renner-Teller substructure have been observed. Using a perturbative analysis, the vibrational frequency has been determined to omega(5) = (710 +/- 4) cm(-1) and the Renner-Teller parameter epsilon(5) is on the order of 3 x 10(-2). PMID- 15783812 TI - Observation of wave packet dichotomy and adiabatic stabilization in an optical waveguide. AB - We report on the first experimental observation of wave packet dichotomy and adiabatic stabilization of light in a periodically bent optical waveguide in analogy with similar behavior of atoms in high-frequency strong laser fields. PMID- 15783813 TI - Laser cooling and magnetic trapping at several tesla. AB - Laser cooling and magnetic trapping of (85)Rb atoms have been performed in extremely strong and tunable magnetic fields, extending these techniques to a new regime and setting the stage for a variety of cold atom and plasma experiments. Using a superconducting Ioffe-Pritchard trap and an optical molasses, 2.4 x 10(7) atoms were laser cooled to the Doppler limit and magnetically trapped at bias fields up to 2.9 T. At magnetic fields up to 6 T, 3 x 10(6) cold atoms were laser cooled in a pulsed loading scheme. These bias fields are well beyond an order of magnitude larger than those in previous experiments. Loading rates, molasses lifetimes, magnetic-trapping times, and temperatures were measured using photoionization and electron detection. PMID- 15783814 TI - Imaging molecular structures by electron diffraction using an intense few-cycle pulse. AB - As an intense few-cycle pulse interacts with an atomic or molecular target, its strong oscillating field may first pull electrons out of the target and subsequently drive them back to scatter on the target. The scattering may occur only a few times or even once during the interaction. This unique property of few cycle pulses enables one to image ultrafast transient structures of matter by the means of pulse-driven electron diffraction. We demonstrated this phenomenon with K(+)(2) via three-dimensional calculations of the time-dependent Schro dinger equation. PMID- 15783815 TI - Temporal shaping of entangled photons. AB - We experimentally demonstrate shaping of the two-photon wave function of entangled-photon pairs, utilizing coherent pulse-shaping techniques. By performing spectral-phase manipulations we tailor the second-order correlation function of the photons exactly like a coherent ultrashort pulse. To observe the shaping we perform sum-frequency generation with an ultrahigh flux of entangled photons. At the appropriate conditions, sum-frequency generation performs as a coincidence detector with an ultrashort response time (approximately 100 fs), enabling a direct observation of the two-photon wave function. This property also enables us to demonstrate background-free, high-visibility two-photon interference oscillations. PMID- 15783816 TI - Photon localization laser: low-threshold lasing in a random amplifying layered medium via wave localization. AB - We demonstrate low-threshold lasing in random amplifying layered medium via photon localization. Lasing is facilitated by resonant excitation of localized modes at the pump laser wavelength, which are peaked deep within the sample with greatly enhanced intensity. Emission occurs into long-lived localized modes overlapping the localized gain region. This mechanism overcomes a fundamental barrier to reducing lasing thresholds in diffusive random lasers, in which multiple scattering restricts the excitation region to the proximity of the sample surface. PMID- 15783817 TI - Azimuthally polarized spatial dark solitons: exact solutions of Maxwell's equations in a Kerr medium. AB - Spatial Kerr solitons, typically associated with the standard paraxial nonlinear Schro dinger equation, are shown to exist to all nonparaxial orders as exact solutions of Maxwell's equations in the presence of the vectorial Kerr effect. More precisely, we prove the existence of azimuthally polarized, spatial, dark soliton solutions of Maxwell's equations, while exact linearly polarized (2 + 1)D solitons do not exist. Our ab initio approach predicts the existence of dark solitons up to an upper value of the maximum field amplitude, corresponding to a minimum soliton width of about one-fourth of the wavelength. PMID- 15783818 TI - Real-space observation of ultraslow light in photonic crystal waveguides. AB - We show the real-space observation of fast and slow pulses propagating inside a photonic crystal waveguide by time-resolved near-field scanning optical microscopy. Local phase and group velocities of modes are measured. For a specific optical frequency we observe a localized pattern associated with a flat band in the dispersion diagram. During at least 3 ps, movement of this field is hardly discernible: its group velocity would be at most c/1000. The huge trapping times without the use of a cavity reveal new perspectives for dispersion and time control within photonic crystals. PMID- 15783803 TI - Search for the flavor-changing neutral current decay B0s--> mu(+) mu(-) in pp Collisions at sqrt[s] = 1.96 TeV with the D0 detector. AB - We present the results of a search for the flavor-changing neutral current decay B(0)(s)--> mu(+) mu(-) using a data set with integrated luminosity of 240 pb(-1) of pp collisions at sqrt[s] = 1.96 TeV collected with the D0 detector in run II of the Fermilab Tevatron collider. We find the upper limit on the branching fraction to be B(B(0)(s)--> mu(+) mu(-)) < or= 5.0 x 10(-7) at the 95% C.L. assuming no contributions from the decay B(0)(d)--> mu(+) mu(-) in the signal region. This limit is the most stringent upper bound on the branching fraction B(0)(s)--> mu(+) mu(-) to date. PMID- 15783819 TI - Channeling chaos by building barriers. AB - Chaotic diffusion often represents a severe obstacle for the setup of experiments, e.g., in fusion plasmas or particle accelerators. We present a complete test of a method of control of Hamiltonian chaos, with both its numerical test and its first experimental realization on a paradigm for wave particle interaction, i.e., a travelling wave tube. The core of our approach is a small apt modification of the system which channels chaos by building barriers to diffusion. Its experimental realization opens the possibility to practically achieve the control of a wide range of systems at a low additional cost of energy. PMID- 15783820 TI - Symmetry breaking via global bifurcations of modulated rotating waves in hydrodynamics. AB - The combined experimental and numerical study finds a complex mechanism of Z(2) symmetry breaking involving global bifurcations for the first time in hydrodynamics. In addition to symmetry breaking via pitchfork bifurcation, the Z(2) symmetry of a rotating wave that occurs in Taylor-Couette flow is broken by a global saddle-node-infinite-period (SNIP) bifurcation after it has undergone a Neimark-Sacker bifurcation to a Z(2)-symmetric modulated rotating wave. Unexpected complexity in the bifurcation structure arises as the curves of cyclic pitchfork, Neimark-Sacker, and SNIP bifurcations are traced towards their apparent merging point. Instead of symmetry breaking due to a SNIP bifurcation, we find a more complex mechanism of Z(2) symmetry breaking involving nonsymmetric two-tori undergoing saddle-loop homoclinic bifurcations and complex dynamics in the vicinity of this global bifurcation. PMID- 15783821 TI - Two-dimensional hard x-ray beam compression by combined focusing and waveguide optics. AB - A two-dimensionally confining x-ray channel waveguide structure is combined with a high gain Kirkpatrick-Baez prefocusing mirror system yielding a hard x-ray beam with a cross section of 25 x 47 nm(2) (FWHM). Unlike the previously employed resonant beam coupling scheme, the incoming beam is coupled in from the front side of the waveguide and the waveguided beam is no longer accompanied by spurious reflected or transmitted beams. The field distribution in the waveguide channel has been calculated numerically. The calculated transmission and far field intensity pattern are in good agreement with the experimental results. PMID- 15783822 TI - Nonlinear harmonic generation in free-electron lasers with helical wigglers. AB - It is widely believed that harmonics are suppressed in helical wigglers. However, linear harmonic generation (LHG) occurs by an azimuthal resonance that excites circularly polarized, off-axis waves, where the hth harmonic varies as exp((ihtheta). Nonlinear harmonic generation (NHG) is driven by bunching at the fundamental and has different properites from LHG. While NHG has been studied in planar wigglers, there has been no analysis of NHG in helical wigglers. The 3D simulation code medusa has been modified for this purpose, and it is shown that NHG is substantial in helical wigglers and that the even and odd harmonics have comparable intensities. PMID- 15783823 TI - Pinning of texture and vortices of the rotating B-like phase of superfluid 3He confined in a 98% aerogel. AB - We have investigated pinning effects on texture and vortices of the B-like phase of superfluid (3)He in a rotating aerogel up to +/-2pi rad/s by cw-NMR. We observed deformation of the NMR spectra in rotation, due to counterflow between the superflow and the normal flow. The average intensity of the counterflow was calculated from the change of NMR spectra. The rotation dependence of the counterflow intensity is similar to the magnetization curve of hard type II superconductors or the counterflow response of (4)He-II in packed powders. This counterflow behavior is in qualitative agreement with a model that vortices are pinned unless the counterflow exceeds a critical velocity v(c). The temperature independence of v(c) suggests that v(c) is associated with the expansion of primordial vortices. PMID- 15783824 TI - Physics of compaction of fine cohesive particles. AB - Fluidized fractal clusters of fine particles display critical-like dynamics at the jamming transition, characterized by a power law relating consolidation stress with volume fraction increment [sigma--(c) proportional, variant(Deltaphi)(beta)]. At a critical stress clusters are disrupted and there is a crossover to a logarithmic law (Deltaphi = nu logsigma--(c)) resembling the phenomenology of soils. We measure lambda identical with- partial differentialDelta(1/phi)/ partial log(sigma--(c) proportional, variant Bo(0.2)(g), where Bo(g) is the ratio of interparticle attractive force (in the fluidlike regime) to particle weight. This law suggests that compaction is ruled by the internal packing structure of the jammed clusters at nearly zero consolidation. PMID- 15783825 TI - Magnetic instabilities in Fe3C cementite particles observed with Fe K-edge x-ray circular dichroism under pressure. AB - The ferromagnetic interstitial iron compound Fe(3)C (cementite) is expected to have Invar properties, whereby a high-moment to low-moment transition should occur when the atomic volume is reduced below a critical value. We, therefore, examine the pressure dependence of the Fe K-edge x-ray magnetic circular dichroism in Fe(3)C at ambient temperature and pressures up to 20 GPa. We find clear evidence for a high-moment to low-moment transition around 10 GPa. PMID- 15783826 TI - Experimental evidence of crystallization pressure inside porous media. AB - Crystallization pressure of salt in porous materials is one of the mechanisms that may induce serious damage, for example, weathering of buildings and monuments of cultural heritage. Since this pressure also causes the solubility of the salt inside a porous material to differ from the bulk solubility, it can be assessed experimentally by measuring the solubility inside the pores. We show that this is possible by NMR, and study Na(2)CO(3) and Na(2)SO(4) in a series of model porous materials. Using the solubility data the crystal-liquid surface energies are estimated as gamma = 0.09 N/m for Na(2)CO(3) . 10H(2)O and gamma = 0.06 N/m for Na(2)SO(4) . 10H(2)O. For pore sizes below about 30 nm, the resulting pressure exceeds the tensile strength of typical building materials (3 MPa). No pressure is induced by the metastable Na(2)SO(4) . 7H(2)O, which suggests for this crystal a value of gamma close to zero. PMID- 15783827 TI - Chain retraction potential in a fixed entanglement network. AB - When a chain, tethered at one end, is immersed in a fixed entanglement network, the mobile tip of the chain encounters an entropic potential barrier that penalizes deep fluctuations needed to bring the tip close to the tethering point. Using the tube model, Doi and Kuzuu [J. Polym. Sci., Polym. Lett. Ed. 18, 775 (1980)] estimated that this potential, which is crucial to describe the rheology of branched polymers in fixed networks and melts, has a quadratic form with a prefactor nu = 1.5. Later calculations based on regular lattices indicated that the potential is nonquadratic, and its steepness depends on the lattice coordination number. In this Letter, we analyze the primitive paths obtained using the bond-fluctuation model for chains with up to 12.5 entanglements. Our simulations confirm a quadratic form for the potential with a prefactor close to the Doi-Kuzuu value, nu approximately 1.5. PMID- 15783828 TI - Sb/Si(111) adsorption: hidden phase transitions behind Langmuir-like isotherms. AB - The experimental study of the thermodynamic and kinetic properties of the Sb/Si(111) interface reveals a surprising behavior: a 2D phase condensates when the Sb coverage increases, indicating strong attractive Sb-Sb interactions, whereas the isotherms present a quasi-Langmuir shape, suggesting that these interactions should be negligible. Ab initio calculations raise this contradiction: while the adsorption site evolves from ternary towards the on-top position with increasing coverage, the character of the Sb-Sb effective interactions changes from repulsive towards attractive, resulting in an almost constant average adsorption energy. A simple (Langmuir) thermodynamic behavior can then be the consequence of a surface phase transition. PMID- 15783829 TI - Two-dimensional carbon incorporation into Si(001): C amount and structure of Si(001)-c(4 x 4). AB - The C amount and the structure of the Si(001)-c(4 x 4) surface is studied using scanning tunneling microscopy (STM) and ab initio calculations. The c(4 x 4) phase is found to contain 1/8 monolayer C (1 C atom in each primitive unit cell). From the C amount and the symmetry of high-resolution STM images, it is inferred that the C atoms substitute the fourth-layer site below the dimer row. We construct a structure model relying on ab initio energetics and STM simulations. Each C atom induces an on-site dimer vacancy and two adjacent rotated dimers on the same dimer row. The c(4 x 4) phase constitutes the subsurface Si(0.875)C(0.125) delta layer with two-dimensionally ordered C atoms. PMID- 15783830 TI - Nanosphere embedding into polymer surfaces: a viscoelastic contact mechanics analysis. AB - Teichroeb and Forrest [Phys. Rev. Lett. 91, 016104 (2003)] image gold nanosphere embedment into a polystyrene surface and imply the existence of a liquid surface layer. We use a viscoelastic contact mechanics model of their results to give a contrary interpretation. The surface interactions between gold and polystyrene and the indentation depth determine the loads on the nanospheres. Using bulk properties, quantitative agreement between the model and the data is obtained, implying little or no depression in the glass temperature or existence of a liquid layer at the polystyrene surface. PMID- 15783831 TI - Probing outer-sphere adsorption of aqueous metal complexes at the oxide-water interface with resonant anomalous x-ray reflectivity. AB - Resonant anomalous x-ray reflectivity near the Pt L(III) edge simultaneously revealed the geometric and spectroscopic structures of Pt(NH(3))(4)(2+) ions adsorbed at the quartz(100)-water interface. The derived Pt geometric subprofile shows two discrete "outer-sphere" adsorbed layers, and the interface-specific x ray absorption edge profile exhibits a significant white-line enhancement compared to the bulk-solution species. PMID- 15783832 TI - Two-photon spectroscopy of core excitons in barium fluoride using synchrotron radiation and laser light. AB - We have conducted two-photon spectroscopy of core excitons in BaF(2). Synchrotron radiation and laser light were used for 5p core-electron excitation and Auger free luminescence was detected as the signal. Two-photon excitation enables access to f and p orbitals that cannot be reached by one-photon excitation of electrons in p orbitals. It has been found that the spin-orbit splittings of 4f and 6p states of the Ba ion in BaF(2) are 0.7 +/- 0.1 and 1.4 +/- 0.1 eV, respectively. PMID- 15783833 TI - Ferromagnetic quantum critical fluctuations in YbRh2(Si0.95Ge0.05)2. AB - The bulk magnetic susceptibility chi(T,B) of YbRh(2)(Si(0.95)Ge(0.05))(2) has been investigated close to the field-induced quantum critical point at B(c) = 0.027 T. For B < or= 0.05 T a Curie-Weiss law with a negative Weiss temperature is observed at temperatures below 0.3 K. Outside this region, the susceptibility indicates ferromagnetic quantum critical fluctuations, chi(T) proportional, variantT-0.6 above 0.3 K. At low temperatures the Pauli susceptibility follows chi(0) proportional, variant(B-B(c))(-0.6) and scales with the coefficient of the T(2) term in the electrical resistivity. The Sommerfeld-Wilson ratio is highly enhanced and increases up to 30 close to the critical field. PMID- 15783834 TI - Wigner crystallization in Na3Cu2O4 and Na8Cu5O10 chain compounds. AB - We report the synthesis of novel edge-sharing chain systems Na(3)Cu(2)O(4) and Na(8)Cu(5)O(10), which form insulating states with commensurate charge order. We identify these systems as one-dimensional Wigner lattices, where the charge order is determined by long-range Coulomb interaction and the number of holes in the d shell of Cu. Our interpretation is supported by x-ray structure data as well as by an analysis of magnetic susceptibility and specific heat data. Remarkably, due to large second neighbor Cu-Cu hopping, these systems allow for a distinction between the (classical) Wigner lattice and the 4k(F) charge-density wave of quantum mechanical origin. PMID- 15783835 TI - Semiclassical analysis of the nonequilibrium local polaron. AB - A resonant level strongly coupled to a local phonon under nonequilibrium conditions is investigated. The nonequilibrium Hartree-Fock approximation is shown to correspond to approximating the steady state density matrix by delta functions at field values to which the local dynamics relaxes in a semiclassical limit. If multiple solutions exist, all are shown to make nonvanishing contributions to physical quantities: multistability does not exist. Departures from equilibrium are shown to produce decoherence, preventing the formation of a polaron feature in the spectral function. The formalism also applies to the nonequilibrium Kondo problem. PMID- 15783836 TI - One-dimensional orbital excitations in vanadium oxides. AB - The d electron orbital is a hidden but important degree of freedom controlling novel properties of transition-metal oxides. A one-dimensional orbital system is especially intriguing due to its enhanced quantum fluctuation. We present a combined experimental and theoretical study on the Raman scattering spectra in perovskite oxides NdVO(3) and LaVO(3) to prove that the quasi-one-dimensional orbital chain described by fermionic pseudospinons bears orbital excitations exchanging occupied orbital states on the neighboring sites, termed a two-orbiton in analogy with two-magnon. PMID- 15783837 TI - Hanle effect driven by weak localization. AB - The influence of weak localization on the Hanle effect in a two-dimensional system with a spin-split spectrum is considered. We show that weak localization drastically changes the dependence of a stationary spin polarization S on an external magnetic field B. In particular, the nonanalytic dependence of S on B is predicted for III-V-based quantum wells grown in the [110] direction and for the [100]-grown quantum wells having equal strengths of Dresselhaus and Bychkov Rashba spin-orbit coupling. It is shown that in a weakly localized regime the components of S are discontinuous at B = 0. At low B, the magnetic field-induced rotation of the stationary polarization is determined by quantum interference effects. This implies that the Hanle effect in such systems is totally driven by weak localization. PMID- 15783838 TI - Effect of spin current on uniform ferromagnetism: domain nucleation. AB - A large spin current applied to a uniform ferromagnet leads to a spin-wave instability as pointed out recently. In this Letter, it is shown that such spin wave instability is absent in a state containing a domain wall, which indicates that nucleation of magnetic domains occurs above a certain critical spin current. This scenario is supported also by an explicit energy comparison of the two states under spin current. PMID- 15783839 TI - Atomic scale conductance induced by single impurity charging. AB - A scanning tunneling microscope was used to probe electron transport through an alkali doped C(60) monolayer crystal on Al(2)O(3) grown by the oxidation of NiAl(110). Each individual alkali atom forms a localized complex with the neighboring C(60) molecules. Charging of the complex induces a substantial rise in the current that persists outside the physical dimensions of the complex. This induction of the current rise is characterized by spatially resolved spectroscopy and mapping of the differential conductance (dI/dV) in the vicinity of the complex. PMID- 15783840 TI - Theory of charge sensing in quantum-dot structures. AB - Charge sensing in quantum-dot structures is studied by an exactly solvable reduced model and numerical density-matrix renormalization-group methods. Charge sensing is characterized by repeated cycling of the occupation of current carrying states due to the capacitive coupling to trap states. In agreement with recent experiments, it results in characteristic asymmetric Coulomb-blockade peaks as well as sawtooth and domelike structures. Temperature introduces asymmetric smearing of these features and correlations in the conductance provide a fingerprint of charge sensing. PMID- 15783841 TI - Spin transition in strongly correlated bilayer two-dimensional electron systems. AB - Using a combination of heat pulse and nuclear magnetic resonance techniques, we demonstrate that the phase boundary separating the interlayer phase coherent quantum Hall effect at nu(T) = 1 in bilayer electron gases from the weakly coupled compressible phase depends upon the spin polarization of the nuclei in the host semiconductor crystal. Our results strongly suggest that, contrary to the usual assumption, the transition is attended by a change in the electronic spin polarization. PMID- 15783842 TI - Nanoscale friction: kinetic friction of magnetic flux quanta and charge density waves. AB - In analogy with the standard macroscopic friction, here we present a comparative study of the friction force felt by moving vortices in superconductors and charge density waves. Using experiments and a model for this data, our observations (1) provide a link between friction at the micro- and macroscopic scales, (2) explain the roundness of the static-kinetic friction transition in terms of thermal fluctuations, particle interactions, and system size (critical-phenomena view), and (3) explain the crossing of the kinetic friction F(k) versus velocity V for our pristine (high density of very weak defects) and our irradiated samples (with lower density of deeper pinning defects). PMID- 15783843 TI - Evidence for charged critical behavior in the pyrochlore superconductor RbOs2O6. AB - We analyze magnetic penetration depth (lambda) data of the recently discovered superconducting pyrochlore oxide RbOs(2)O(6). Our results strongly suggest that in RbOs(2)O(6) charged critical fluctuations dominate the temperature dependence of lambda near T(c). This is in contrast with the mean-field behavior observed in conventional superconductors and the uncharged critical behavior found in nearly optimally doped cuprate superconductors. However, this finding agrees with the theoretical predictions for charged criticality and the charged criticality observed in underdoped YBa(2)Cu(3)O(6.59). PMID- 15783844 TI - In situ measurement of self-heating in intrinsic tunneling spectroscopy. AB - Using advanced sample engineering we performed simultaneous measurements of interlayer tunneling characteristics and in situ monitoring of temperature in Bi(2)Sr(2)CaCu(2)O(8 + delta) (Bi-2212) mesas. Together with a systematic study of size dependence of interlayer tunneling, this allowed unambiguous discrimination between artifacts of self-heating and gaps in the electronic spectra of Bi-2212. Such a confident spectroscopic information, which is not affected by self-heating or surface deterioration, was obtained for the first time for a high-T(c) superconductor. We also derived general expressions and formulated main principles of self-heating valid for a large variety of materials. PMID- 15783845 TI - Surface superconductivity of dirty two-band superconductors: applications to MgB2. AB - The minimal magnetic field H(c2) destroying superconductivity in the bulk of a superconductor is smaller than the magnetic field H(c3) needed to destroy surface superconductivity if the surface of a superconductor coincides with one of the crystallographic planes and is parallel to the external magnetic field. While for a dirty single-band superconductor the ratio of H(c3) to H(c2) is a universal temperature-independent constant 1.6946, for dirty two-band superconductors this is not the case. I show that in the latter case the interaction of the two bands leads to a novel scenario with the ratio H(c3)/H(c2) varying with temperature and taking values larger and smaller than 1.6946. The results are applied to MgB(2) and compared with recent experiments (A. Rydh, cond-mat/0307445). PMID- 15783846 TI - Competitive localization of vortex lines and interacting bosons. AB - We present a theory for the localization of three-dimensional vortex lines or two dimensional bosons with a short-ranged repulsive interaction which are competing for a single columnar defect or potential well. For two vortices we use a necklace model approach to find a new kind of delocalization transition between two different states with a single bound particle. This exchange-delocalization transition is characterized by the onset of vortex exchange on the defect for sufficiently weak vortex-vortex repulsion or sufficiently weak binding energy corresponding to high temperature. We calculate the transition point and order of the exchange-delocalization transition. A generalization of this transition to an arbitrary vortex number is proposed. PMID- 15783847 TI - Low-energy properties of aperiodic quantum spin chains. AB - We investigate the low-energy properties of antiferromagnetic quantum XXZ spin chains with couplings following two-letter aperiodic sequences, by an adaptation of the Ma-Dasgupta-Hu renormalization-group method. For a given aperiodic sequence, we argue that, in the easy-plane anisotropy regime, intermediate between the XX and Heisenberg limits, the general scaling form of the thermodynamic properties is essentially given by the exactly known XX behavior, providing a classification of the effects of aperiodicity on XXZ chains. As representative illustrations, we present analytical and numerical results for the low-temperature thermodynamics and the ground-state correlations for couplings following the Fibonacci quasiperiodic structure and a binary Rudin-Shapiro sequence, whose geometrical fluctuations are similar to those induced by randomness. PMID- 15783848 TI - True single domain and configuration-assisted switching of submicron Permalloy dots observed by electron holography. AB - The switching behavior of submicron circular Permalloy nanomagnets has been investigated. Electron holography provides a magnetic resolution of down to 10 nm. This allows us to observe in detail the switching and to measure the induction within single nanodots with diameters down to 150 nm at a thickness of 6 nm. Particles of these dimensions show a single domain state during the whole switching process which takes place at external fields of only a few 100 A/m. For larger or thicker particles the magnetization reversal runs via the formation of a C state or an intermediate vortex state. PMID- 15783849 TI - NMR as a probe of the relaxation of the magnetization in magnetic molecules. AB - We investigate the time autocorrelation of the molecular magnetization M(t) for three classes of magnetic molecules (antiferromagnetic rings, grids, and nanomagnets), in contact with the phonon heat bath. For all three classes, we find that the exponential decay of the fluctuations of M(t) is characterized by a single characteristic time tau(T,B) for not too high temperature T and field B. This is reflected in a nearly single-Lorentzian shape of the spectral density of the fluctuations. We show that such fluctuations are effectively probed by NMR, and that our theory explains the recent phenomenological observation by Baek et al. [Phys. Rev. B 70, 134434 (2004)] that the Larmor-frequency dependence of 1/T(1) data in a large number of AFM rings fits to a single-Lorentzian form. PMID- 15783850 TI - Glassy properties of the Kawasaki dynamics of two-dimensional ferromagnets. AB - We study numerically the Kawasaki dynamics of the 2D Ising model. At large time we recover the coarsening growth as l(c)(t) proportional t(1/3). At shorter time however, the system enters a metastable glassy regime that displays an extremely slow growth and nontrivial violations of the fluctuation-dissipation theorem similar to those observed in spin glasses: this is one of the simplest systems in which such violations occur. We also consider Potts models, where a similar behavior is observed, and the model of Shore and Sethna where the domain growth is also slow, but where violations of the fluctuation-dissipation theorem are trivial. We finally comment on these violations in the context of activated coarsening, and on similarities and differences with the glass transition phenomenology. PMID- 15783851 TI - Optical magnetoelectric effect in a submicron patterned magnet. AB - The optical magnetoelectric effect, which is a nonreciprocal directional dichroic response, has been demonstrated in a submicron patterned magnet by monitoring the diffracted visible or near-infrared light intensity. An artificial magnetic superstructure is composed of chevron shaped ">" islands made of the ferromagnetic permalloy Ni(80)Fe(20) with a pitch of 1 microm on silicon substrate, in which both space inversion and time reversal symmetry are broken simultaneously. On the basis of the light-polarization angle and magnetic field H dependence, and also comparing the results with the those of the submicron square patterns, we show that the optical magnetoelectric effect emerges as the finite change (approximately 10(-3) at room temperature in H of 500 Oe) of the diffracted intensity. PMID- 15783853 TI - Liquid-vapor interface in liquid binary alloys: an ab initio molecular dynamics study. AB - We report the results of an ab initio molecular dynamics simulation of the liquid vapor interface of two binary liquid alloys, Na(0.3)K(0.7) and Li(0.4)Na(0.6), whose bulk behavior exhibits rather differing ordering tendencies. The study has been performed using samples of 2000 and 3000 particles, respectively, in a slab geometry with periodic boundary conditions. In both cases, the total ionic density distributions along the normal to the interface display some layering with a virtually pure monolayer of the lower surface tension component located outermost at the interface. However, the two systems behave very differently below the interface which can be accounted for by their different ordering tendencies in the bulk. PMID- 15783852 TI - Quantum oscillation of Hall resistance in the extreme quantum limit of an organic conductor (TMTSF)2ClO4. AB - We report resistance and magnetic torque experiments under a high magnetic field up to 45 T in a three dimensional quantum Hall (QH) system (TMTSF)(2)ClO(4), where TMTSF = tetramethyltetraselenafulvalene. The Hall resistance shows huge oscillations accompanied with sign reversal after the final QH state, where the Landau level filling factor is unity, is removed above 26 T. The magnetic torque also oscillates with the field. The results suggest that a novel quantum state, where the character of the carriers periodically changes with the field, is stabilized in the extreme quantum limit. PMID- 15783855 TI - What aligns liquid crystals on solid substrates? The role of surface roughness anisotropy. AB - The mechanism responsible for liquid crystal (LC) alignment on mechanically buffed or UV exposed polymer films is poorly understood. A comprehensive study of LC alignment on variously prepared substrates unequivocally shows that the anisotropy in the surface roughness of the substrate completely determines the direction of LC alignment. In all the cases studied, including those where an anchoring transition occurs with temperature, the LC director (re)aligns in the directions of low roughness. PMID- 15783854 TI - Origin of the short-range, strong repulsive force between ionic surfactant layers. AB - We study the electrostatic interaction between two ionic surfactant layers by performing molecular dynamic simulations of salt-free thin water films coated by surfactants (Newton black films). We find a strong exponentially decaying short range repulsion not explained by classical Poisson-Boltzmann theory. This electrostatic force is shown to be mainly due to the anomalous dielectric response of water near charged surfactant layers. This result clarifies the much debated physical mechanism underlying the controversial "hydration forces" observed in experiments. In the case of ionic thin films, the "hydration forces" can be identified with the electrostatic forces induced by the layers of highly polarized water originated at the interfaces. PMID- 15783856 TI - Propagation and relaxation of tension in stiff polymers. AB - We present a unified theory for the longitudinal dynamic response of a stiff polymer in solution to various external perturbations (mechanical excitations, hydrodynamic flows, electrical fields, temperature quenches, etc.) that can be represented as sudden changes of ambient/boundary conditions. The theory relies on a comprehensive analysis of the nonequilibrium propagation and relaxation of backbone stresses in a wormlike chain. We recover and substantially extend previous results based on heuristic arguments. New experimental implications are pointed out. PMID- 15783857 TI - Morphogen gradient from a noisy source. AB - We investigate the effect of time-dependent noise on the shape of a morphogen gradient in a developing embryo. Perturbation theory is used to calculate the deviations from deterministic behavior in a simple reaction-diffusion model of robust gradient formation, and the results are confirmed by numerical simulation. It is shown that such deviations can disrupt robustness for sufficiently high noise levels, and the implications of these findings for more complex models of gradient-shaping pathways are discussed. PMID- 15783858 TI - Protein structural change upon ligand binding: linear response theory. AB - A simple formula based on linear response theory is proposed to explain and predict the structural change of proteins upon ligand binding. By regarding ligand binding as an external perturbation, the structural change as a response is described by atomic fluctuations in the ligand-free form and the protein ligand interactions. The results for three protein systems of various sizes are consistent with the observations in the crystal structures, confirming the validity of the linear relationship between the equilibrium fluctuations and the structural change upon ligand binding. PMID- 15783860 TI - Scaling of dynamics with the range of interaction in short-range attractive colloids. AB - We numerically study the dependence of the dynamics on the range of interaction Delta for the short-range square well potential. We find that, for small Delta, dynamics scale exactly in the same way as thermodynamics, both for Newtonian and Brownian microscopic dynamics. For interaction ranges from a few percent down to the Baxter limit, the relative location of the attractive-glass line and the liquid-gas line does not depend on Delta. This proves that, in this class of potentials, disordered arrested states (gels) can be generated only as a result of a kinetically arrested phase separation. PMID- 15783859 TI - Designing pattern-recognition surfaces for selective adsorption of copolymer sequences using lattice monte carlo simulation. AB - We describe a simulation method to design surfaces for recognizing specific monomer sequences in copolymers. We fix the monomer sequence statistics of the AB copolymers on a surface containing two types of sites and allow the simulation to iterate towards an optimal surface pattern that can recognize and selectively adsorb the sequence in the copolymer. During the simulation the surface pattern is designed by switching identities of two randomly picked sites. For copolymers with less blocky sequences the designed surfaces recognize the correct sequence well when the segment-surface interactions dominate over the intersegment interactions. For copolymers with more blocky sequences recognition is good when the segment-surface interactions are only slightly stronger than the intersegment interactions. PMID- 15783861 TI - Single-molecule pump-probe detection resolves ultrafast pathways in individual and coupled quantum systems. AB - We report the first experimental study of individual molecules with femtosecond time resolution using a novel ultrafast single-molecule pump-probe method. A wide range of relaxation times from below 100 up to 400 fs is found, revealing energy redistribution over different vibrational modes and phonon coupling to the nanoenvironment. Addressing quantum-coupled molecules we find longer decay times, pointing towards inhibited intramolecular decay due to delocalized excitation. Interestingly, each individual system shows discrete jumps in femtosecond response, reflecting sudden breakup of the coupled superradiant state. PMID- 15783862 TI - Wetting, drying, and layering of colloid-polymer mixtures at porous interfaces. AB - The influence of interface porosity on the wetting properties of colloid-polymer mixtures is studied within density functional theory for the Asakura-Oosawa-Vrij model at the surface of a quenched hard-sphere matrix. While the porosity hardly changes the location of the transition from partial to complete wetting at colloidal bulk gas-liquid coexistence, the onset of wetting, as signaled by the first discontinuous layering transition, can be efficiently controlled by tailoring the porosity. We furthermore find that the penetrability of the porous interface induces complete drying into the matrix upon approaching capillary coexistence. PMID- 15783863 TI - Comment on "quantum entropy and special relativity". PMID- 15783865 TI - Comment on "thermal stability and electronic structure of atomically uniform Pb films on Si(111)". PMID- 15783867 TI - Justification of c-number substitutions in bosonic Hamiltonians. AB - The validity of substituting a c-number z for the k=0 mode operator a(0) is established rigorously in full generality, thereby verifying one aspect of Bogoliubov's 1947 theory. This substitution not only yields the correct value of thermodynamic quantities such as the pressure or ground state energy, but also the value of |z|(2) that maximizes the partition function equals the true amount of condensation in the presence of a gauge-symmetry-breaking term. This point had previously been elusive. PMID- 15783868 TI - Equivalence of Bose-Einstein condensation and symmetry breaking. AB - Based on a classic paper by Ginibre [Commun. Math. Phys. 8, 26 (1968)] it is shown that, whenever Bogoliubov's approximation, that is, the replacement of a(0) and a*(0) by complex numbers in the Hamiltonian, asymptotically yields the right pressure, it also implies the asymptotic equality of |a(0)|(2)/V and a*(0)a(0)/V in symmetry breaking fields, irrespective of the existence or absence of Bose Einstein condensation. Because the former was proved by Ginibre to hold for absolutely integrable superstable pair interactions, the latter is equally valid in this case. Apart from Ginibre's work, our proof uses only a simple convexity inequality due to Griffiths. PMID- 15783869 TI - Fermionic atoms in a three dimensional optical lattice: observing Fermi surfaces, dynamics, and interactions. AB - We have studied interacting and noninteracting quantum degenerate Fermi gases in a three-dimensional optical lattice. We directly image the Fermi surface of the atoms in the lattice by turning off the optical lattice adiabatically. Because of the confining potential, gradual filling of the lattice transforms the system from a normal state into a band insulator. The dynamics of the transition from a band insulator to a normal state is studied, and the time scale is measured to be an order of magnitude larger than the tunneling time in the lattice. Using a Feshbach resonance, we increase the interaction between atoms in two different spin states and dynamically induce a coupling between the lowest energy bands. We observe a shift of this coupling with respect to the Feshbach resonance in free space which is anticipated for strongly confined atoms. PMID- 15783870 TI - Observations on sound propagation in rapidly rotating Bose-Einstein condensates. AB - Repulsive laser potential pulses applied to vortex lattices of rapidly rotating Bose-Einstein condensates create propagating density waves which we have observed experimentally and modeled computationally to high accuracy. We have observed a rich variety of dynamical phenomena ranging from interference effects and shock wave formation to anisotropic sound propagation. PMID- 15783871 TI - Iterative optimization of quantum error correcting codes. AB - We introduce a convergent iterative algorithm for finding the optimal coding and decoding operations for an arbitrary noisy quantum channel. This algorithm does not require any error syndrome to be corrected completely, and hence also finds codes outside the usual Knill-Laflamme definition of error correcting codes. The iteration is shown to improve the figure of merit "channel fidelity" in every step. PMID- 15783872 TI - Nonergodicity of blinking nanocrystals and other Levy-walk processes. AB - We investigate the nonergodic properties of blinking nanocrystals modeled by a Levy-walk stochastic process. Using a nonergodic mean field approach we calculate the distribution functions of the time averaged intensity correlation function. We show that these distributions are not delta peaked on the ensemble average correlation function values; instead they are W or U shaped. Beyond blinking nanocrystals our results describe ergodicity breaking in systems modeled by Levy walks , for example, certain types of chaotic maps and spin dynamics to name a few. PMID- 15783873 TI - Thermodynamic theory of incompressible hydrodynamics. AB - The grand potential for open systems describes thermodynamics of fluid flows at low Mach numbers. A new system of reduced equations for the grand potential and the fluid momentum is derived from the compressible Navier-Stokes equations. The incompressible Navier-Stokes equations are the quasistationary solution to the new system. It is argued that the grand canonical ensemble is the unifying concept for the derivation of models and numerical methods for incompressible fluids, illustrated here with a simulation of a minimal Boltzmann model in a microflow setup. PMID- 15783874 TI - Lorentz violation in supersymmetric field theories. AB - We construct supersymmetric Lorentz violating operators for matter and gauge fields. We show that in the supersymmetric standard model the lowest possible dimension for such operators is five, and therefore they are suppressed by at least one power of an ultraviolet energy scale, providing a possible explanation for the smallness of Lorentz violation and its stability against radiative corrections. Supersymmetric Lorentz noninvariant operators do not lead to modifications of dispersion relations at high energies thereby escaping constraints from astrophysical searches for Lorentz violation. PMID- 15783875 TI - Measurement of neutrino oscillation with KamLAND: evidence of spectral distortion. AB - We present results of a study of neutrino oscillation based on a 766 ton/year exposure of KamLAND to reactor antineutrinos. We observe 258 nu (e) candidate events with energies above 3.4 MeV compared to 365.2+/-23.7 events expected in the absence of neutrino oscillation. Accounting for 17.8+/-7.3 expected background events, the statistical significance for reactor nu (e) disappearance is 99.998%. The observed energy spectrum disagrees with the expected spectral shape in the absence of neutrino oscillation at 99.6% significance and prefers the distortion expected from nu (e) oscillation effects. A two-neutrino oscillation analysis of the KamLAND data gives Deltam(2)=7.9(+0.6)(-0.5)x10(-5) eV(2). A global analysis of data from KamLAND and solar-neutrino experiments yields Deltam(2)=7.9(+0.6)(-0.5)x10(-5) eV(2) and tan((2)theta=0.40(+0.10)( 0.07), the most precise determination to date. PMID- 15783877 TI - Measurement of the transverse beam spin asymmetry in elastic electron-proton scattering and the inelastic contribution to the imaginary part of the two-photon exchange amplitude. AB - We report on a measurement of the asymmetry in the scattering of transversely polarized electrons off unpolarized protons, A( perpendicular), at two Q2 values of 0.106 and 0.230 (GeV/c)(2) and a scattering angle of 30 degrees N-->nN processes in a finite nucleus framework. The relationship between the intrinsic Lambda asymmetry parameter a(Lambda) and the asymmetry a(M)(Lambda) accessible in experiments is discussed. A strong dependence of a(M)(Lambda) on nucleon final state interactions and detection threshold is obtained. Our results for a(M)(Lambda) are consistent with (11)(Lambda)B--> and (12)(Lambda)C--> data but disagree with observations in (5)(Lambda)H-->e. PMID- 15783885 TI - Direct measurement of the 14N(p,gamma)15O S factor. AB - The 14N(p,gamma)15O reaction regulates the rate of energy generation in the stellar CN cycle. Because discrepancies have been found in the analysis and interpretation of previous capture data, we have measured the 14N(p,gamma)15O excitation function for energies in the range E(lab)(p)=155-524 keV. Fits of these data using R-matrix theory yield a value for the S factor at zero energy of 1.68+/-0.09(stat)+/-0.16(syst) keV b, which is significantly smaller than the previous result. The corresponding reduction in the stellar reaction rate for 14N(p,gamma)15O has a number of interesting consequences, including an impact on estimates for the age of the Galaxy derived from globular clusters. PMID- 15783884 TI - Neutron spectroscopic factors in 9Li from 2H(8Li,p)9Li. AB - We have studied the 2H(8Li,p)9Li reaction to obtain information on the spins, parities, and single-neutron spectroscopic factors for states in 9Li, using a radioactive 8Li beam. The deduced properties of the lowest three states are compared to the predictions of a number of calculations for the structure of 9Li. The results of ab initio quantum Monte Carlo calculations are in good agreement with the observed properties. PMID- 15783886 TI - ac electric trap for ground-state molecules. AB - We here report on the realization of an electrodynamic trap, capable of trapping neutral atoms and molecules in both low-field and high-field seeking states. Confinement in three dimensions is achieved by switching between two electric field configurations that have a saddle point at the center of the trap, i.e., by alternating a focusing and a defocusing force in each direction. The ac trapping of 15ND(3) molecules is experimentally demonstrated, and the stability of the trap is studied as a function of the switching frequency. A 1 mK sample of 15ND(3) molecules in the high-field seeking component of the |J,K=|1,1 level, the ground state of para-ammonia, is trapped in a volume of about 1 mm(3). PMID- 15783887 TI - Simple route to strong-field coherent control. AB - Coherent-control schemes to manipulate weak-field interactions are generally invalid at stronger fields, since strong-field interactions are accompanied by level power broadenings and level shifts that usually elude simple analytical treatments. Here we show that a broad subgroup of weak-field solutions (those with real fields, i.e., fields with only one quadrature in the complex plane) can be extended to the strong-field regime while retaining their properties. The salient feature of these fields is a symmetry that cancels out power broadening effects. Such fields can be generated from ultrashort coherent pulses or from incoherent broadband down-converted light. Weak-field coherent-control approaches based on these solutions can therefore be extended to the strong-field regime as we demonstrate in a two-photon absorption experiment in atomic cesium. PMID- 15783882 TI - Measurement of the 3He(e,e'p)pn reaction at high missing energies and momenta. AB - Results of the Jefferson Lab Hall A quasielastic 3He(e,e'p)pn measurements are presented. These measurements were performed at fixed transferred momentum and energy, q=1502 MeV/c and omega=840 MeV, respectively, for missing momenta p(m) up to 1 GeV/c and missing energies in the continuum region, up to pion threshold; this kinematic coverage is much more extensive than that of any previous experiment. The cross section data are presented along with the effective momentum density distribution and compared to theoretical models. PMID- 15783888 TI - Mapping attosecond electron wave packet motion. AB - Attosecond pulses are produced when an intense infrared laser pulse induces a dipole interaction between a sublaser cycle recollision electron wave packet and the remaining coherently related bound-state population. By solving the time dependent Schrodinger equation we show that, if the recollision electron is extracted from one or more electronic states that contribute to the bound-state wave packet, then the spectrum of the attosecond pulse is modulated depending on the relative motion of the continuum and bound wave packets. When the internal electron and recollision electron wave packet counterpropagate, the radiation intensity is lower. We show that we can fully characterize the attosecond bound state wave packet dynamics. We demonstrate that electron motion from a two-level molecule with an energy difference of 14 eV, corresponding to a period of 290 asec, can be resolved. PMID- 15783889 TI - Photoassociative spectroscopy at long range in ultracold strontium. AB - We report photoassociative spectroscopy of 88Sr(2) in a magneto-optical trap operating on the 1S0-->3P1 intercombination line at 689 nm. Photoassociative transitions are driven with a laser red detuned by 600-2400 MHz from the 1S0- >1P1 atomic resonance at 461 nm. Photoassociation takes place at extremely large internuclear separation, and the photoassociative spectrum is strongly affected by relativistic retardation. A fit of the transition frequencies determines the 1P1 atomic lifetime (tau=5.22+/-0.03 ns) and resolves a discrepancy between experiment and recent theoretical calculations. PMID- 15783878 TI - Centrality dependence of charm production from a measurement of single electrons in Au+Au collisions at sqrt[s(NN)]=200 GeV. AB - The PHENIX experiment has measured midrapidity transverse momentum spectra (0.4. Explicit time-dependent nuclear polarization expressions for pulsed rotational excitation are presented for the case of one nuclear spin I, and for the case of two nuclear spins I1 and I2 in the hierarchical coupling limit. Photodissociation of the polarized molecules allows the production of polarized atoms, on short time scales if pulsed, at densities close to the parent molecule density. PMID- 15783879 TI - Nuclear modification factors for hadrons at forward and backward rapidities in deuteron-gold collisions at sqrt[s(NN)]=200 GeV. AB - We report on charged hadron production in deuteron-gold reactions at sqrt[s(NN)]=200 GeV. Our measurements in the deuteron direction cover 1.4 or =22) dilute 2D GaAs hole system using a standard carrier heating technique. In the still poorly understood metallic state, we observe that P(T) changes from P(T) approximately T5 to P(T) approximately T7 above 100 mK, indicating a crossover from screened piezoelectric (PZ) coupling to screened deformation potential (DP) coupling for hole-phonon scattering. Quantitative comparison with theory shows that the long range PZ coupling between holes and phonons has the expected magnitude; however, in the metallic state, the short range DP coupling between holes and phonons is almost 20 times stronger than expected from theory. The density dependence of P(T) shows that it is easier to cool low-density 2D holes in GaAs than higher density 2D hole systems. PMID- 15783912 TI - Kondo screening cloud around a quantum dot: large-scale numerical results. AB - Measurements of the persistent current in a ring containing a quantum dot would afford a unique opportunity to finally detect the elusive Kondo screening cloud. We present the first large-scale numerical results on this controversial subject using exact diagonalization and density matrix renormalization group (RG). These extremely challenging numerical calculations confirm RG arguments for weak to strong coupling crossover with varying ring length and give results on the universal scaling functions. We also study, analytically and numerically, the important and surprising effects of particle-hole symmetry breaking. PMID- 15783913 TI - Ruderman-Kittel-Kasuya-Yosida and magnetic-field interactions in coupled Kondo quantum dots. AB - We investigate theoretically the transport properties of two independent artificial Kondo impurities. They are coupled together via a tunable Ruderman Kittel-Kasuya-Yosida (RKKY) interaction. For strong enough antiferromagnetic RKKY interaction, the impurity density of states increases with the applied in-plane magnetic-field. This effect can be used to distinguish between antiferromagnetic and ferromagnetic RKKY interactions. These results may be relevant to explain some features of recent experiments by Craig et al. [Science 304, 565 (2004)]. PMID- 15783914 TI - Temperature dependence of the tunneling amplitude between quantum hall edges. AB - Recent experiments have studied the tunneling current between the edges of a fractional quantum Hall liquid as a function of temperature and voltage. The results of the experiment are puzzling because at "high" temperature (600-900 mK) the behavior of the tunneling conductance is consistent with the theory of tunneling between chiral Luttinger liquids, but at low temperature it strongly deviates from that prediction dropping to zero with decreasing temperature. In this Letter we suggest a possible explanation of this behavior in terms of the strong temperature dependence of the tunneling amplitude. PMID- 15783915 TI - Electron-phonon interaction and transport in semiconducting carbon nanotubes. AB - We calculate the electron-phonon scattering and binding in semiconducting carbon nanotubes, within a tight-binding model. The mobility is derived using a multiband Boltzmann treatment. At high fields, the dominant scattering is interband scattering by LO phonons corresponding to the corners K of the graphene Brillouin zone. The drift velocity saturates at approximately half the graphene Fermi velocity. The calculated mobility as a function of temperature, electric field, and nanotube chirality are well reproduced by a simple interpolation formula. Polaronic binding give a band-gap renormalization of approximately 70 meV, an order of magnitude larger than expected. Coherence lengths can be quite long but are strongly energy dependent. PMID- 15783916 TI - Fractional quantum Hall states of atoms in optical lattices. AB - We describe a method to create fractional quantum Hall states of atoms confined in optical lattices. We show that the dynamics of the atoms in the lattice is analogous to the motion of a charged particle in a magnetic field if an oscillating quadrupole potential is applied together with a periodic modulation of the tunneling between lattice sites. In a suitable parameter regime the ground state in the lattice is of the fractional quantum Hall type, and we show how these states can be reached by melting a Mott-insulator state in a superlattice potential. Finally, we discuss techniques to observe these strongly correlated states. PMID- 15783917 TI - Electron interactions and transport between coupled quantum Hall edge states. AB - We examine the effects of electron-electron interactions on transport between edge states in a multilayer integer quantum Hall system. The edge states of such a system, coupled by interlayer tunneling, form a two-dimensional, chiral metal at the sample surface. We calculate the temperature-dependent conductivity and the amplitude of conductance fluctuations in this chiral metal, treating Coulomb interactions and disorder exactly in the weak-tunneling limit. We find that the conductivity increases with increasing temperature, as observed in recent experiments, and we show that the correlation length characterizing conductance fluctuations varies inversely with temperature. PMID- 15783918 TI - Transport spectroscopy of Kondo quantum dots coupled by RKKY interaction. AB - We develop the theory of conductance of a quantum dot which carries a spin and is coupled via RKKY interaction to another spin-carrying quantum dot. The found dependence of the differential conductance on the bias and magnetic field at a fixed RKKY interaction strength may allow one to distinguish between the possible ground states of the system. Transitions between the ground states are achieved by tuning the RKKY interaction, and the nature of these transitions can be extracted from the temperature dependence of the linear conductance. The feasibility of the corresponding measurements is evidenced by recent experiments by Craig et al. PMID- 15783919 TI - Noise and full counting statistics of incoherent multiple Andreev reflection. AB - We present a general theory for the full counting statistics of multiple Andreev reflections in incoherent superconducting-normal-superconducting contacts. The theory, based on a stochastic path integral approach, is applied to a superconductor-double-barrier system. It is found that all cumulants of the current show a pronounced subharmonic gap structure at voltages V=2Delta/en. For low voltages V< or =3. We show that this low-voltage result holds for a large class of incoherent superconducting-normal-superconducting contacts. PMID- 15783920 TI - Uniformly frustrated XY model without a vortex-pattern ordering. AB - The uniformly frustrated XY model with f=1/3 on a dice lattice is shown to possess an accidental degeneracy of its ground states so well developed that the difference between the free energies of fluctuations does not lead to the stabilization of a particular vortex pattern down to zero temperature. Nonetheless, at low temperatures the system is characterized by a finite helicity modulus whose vanishing (at a finite temperature) is related to the dissociation of half-vortex pairs. PMID- 15783921 TI - Generalized anisotropic scaling theory and the transverse meissner transition. AB - We consider a depinning transition in vortex systems with columnar disorder and tilted applied magnetic fields. From scaling arguments and Monte Carlo simulations, we find that this transverse Meissner transition is governed by a fixed point which is anisotropic in all three directions. This generalization of conventional anisotropic scaling means that the correlation length in different directions diverges with different rates, and we derive exact results for the anisotropy exponents. We make predictions which can be tested in experiments on superconductors with columnar disorder. PMID- 15783923 TI - Thermal conductivity of quasi-one-dimensional antiferromagnetic spin-chain materials. AB - We study heat transport in quasi-one-dimensional spin-chain systems by considering the model of one-dimensional bosonic spin excitations interacting with three-dimensional phonons and impurities in the limit of weak spin-lattice coupling and fast spin excitations. A combined effect of the phonon and impurity scatterings yields the following spin-boson thermal conductivity behavior: kappa(s) proportional to T2 at low, kappa(s) proportional to T-1 at intermediate, and kappa(s)= const at higher temperatures. Our results agree well with the existing experimental data for Sr2CuO3. We predict an unusual dependence on the impurity concentration for a number of observables and propose further experiments. PMID- 15783922 TI - Macroscopic quantum tunneling in d-wave YBa2Cu3O7-delta Josephson junctions. AB - The escape rate from the zero voltage state in a superconducting Josephson junction (JJ) is determined by the temperature, but it saturates at low temperature due to macroscopic quantum tunneling (MQT). Complications due to d wave symmetry in a high temperature superconductor, like low energy quasiparticles and an unconventional current-phase relation, may influence the escape rate. We report, for the first time to our knowledge, the observation of MQT in a YBa(2)Cu(3)O(7-delta) grain boundary biepitaxial JJ. This proves that dissipation can be significantly reduced by a proper junction configuration, which is of significance for quantum coherence. PMID- 15783925 TI - Roundabout relaxation: collective excitation requires a detour to equilibrium. AB - Relaxation to equilibrium after strong and collective excitation is studied by using a Hamiltonian dynamical system of a one-dimensional XY model. After an excitation of a domain of K elements, the excitation is concentrated to fewer elements, which are made farther away from equilibrium, and the excitation intensity increases logarithmically with K. Equilibrium is reached only after taking this roundabout route, with the time for relaxation diverging asymptotically as Kgamma with gamma approximately 4.2. PMID- 15783924 TI - General features of photoinduced spin dynamics in ferromagnetic and ferrimagnetic compounds. AB - Ultrafast photoinduced spin dynamics has been investigated by time-resolved magneto-optical Kerr spectroscopy for various ferromagnetic and ferrimagnetic compounds: FeCr2S4, CoCr2S4, CuCr2Se4, CdCr2Se4, La0.6Sr0.4MnO3, and SrRuO3. The temporal demagnetization process, which is observed commonly for all the compounds, essentially consists of two components: One is an instantaneous change which originates perhaps from multiple emissions of magnetic excitations during nonradiative decay of photoexcited carriers, and the other is a delayed response due to thermalization of the spin system. The time constant of the delayed change depends strongly on materials and is scaled with the magnetocrystalline anisotropy, indicating that spin-orbit coupling is a dominant interaction for this process. PMID- 15783926 TI - Revealing antiferromagnetic order of the Fe monolayer on W(001): spin-polarized scanning tunneling microscopy and first-principles calculations. AB - We prove that the magnetic ground state of a single monolayer Fe on W(001) is c(2x2) antiferromagnetic, i.e., a checkerboard arrangement of antiparallel magnetic moments. Real space images of this magnetic structure have been obtained with spin-polarized scanning tunneling microscopy. An out-of-plane easy magnetization axis is concluded from measurements in an external magnetic field. The magnetic ground state and anisotropy axis are explained based on first principles calculations. PMID- 15783927 TI - Orbital order and ferrimagnetic properties of Sr8CaRe3Cu4O24. AB - By means of the LSDA+U (local spin density approximation plus Hubbard U) method and the Green function method, we investigate the electronic and magnetic properties of the new material of Sr8CaRe3Cu4O24. Our LSDA+U calculation shows that this system is an insulator of ferrimagnetism with a net magnetic moment of 1.01 micro(B)/f.u., which is in good agreement with the experiment. It is the nonmagnetic Re atoms that induce an orbital order of d electrons of Cu atoms, which is responsible for the strong exchange interaction and the high magnetic transition temperature. Based on the LSDA+U results, we introduce an effective model for the spin degrees of freedom and investigate the finite-temperature properties by the Green function method. The obtained results are consistent with the experimental results, indicating that the spin-alternating Heisenberg model is suitable for this compound. PMID- 15783928 TI - Optical polarization driven giant relief modulation in amorphous chalcogenide glasses. AB - We report on the observation of giant relief modulation in amorphous chalcogenide As2S3 glass under polarization modulated near band gap light illumination. We show that these periodic modulations are created as a result of the photoinduced mass transport despite the very low and uniform intensity light illumination. A phenomenological model, based on a photoinduced alignment of anisotropic microvolumes within the glass, is proposed to explain the phenomenon. PMID- 15783929 TI - Polarization dependence of the optical absorption of single-walled carbon nanotubes. AB - Anisotropic optical absorption properties of single-walled carbon nanotubes (SWNTs) are determined from a vertically aligned SWNT film for 0.5-6 eV. Absorption peaks at 4.5 and 5.25 eV are found to exhibit remarkable polarization dependence and have relevance to optical properties of graphite. A method for determining a nematic order parameter for an aligned SWNT film based on the collinear absorption peak at 4.5 eV is presented, followed by the determination of the optical absorption cross section. PMID- 15783930 TI - Room-temperature ordered photon emission from multiexciton states in single CdSe core-shell nanocrystals. AB - We report room-temperature ordered multiphoton emission from multiexciton states of single CdSe(CdZnS) core(-shell) colloidal nanocrystals (NCs) that are synthesized by wet chemical methods. Spectrally and temporally resolved measurements of biexciton and triexciton emission from single NCs are also presented. A simple four level system models the results accurately and provides estimates for biexciton and triexciton radiative lifetimes and quantum yields. PMID- 15783931 TI - Reflectance anisotropy spectra of the diamond (100)-(2x1) surface: evidence of strongly bound surface state excitons. AB - We compare the results of ab initio calculations with measured reflection anisotropy spectra and show that strongly bound surface-state excitons occur on the clean diamond (100) surface. These excitons are found to have a binding energy close to 1 eV, the strongest ever observed at a semiconductor surface. Important electron-hole interaction effects on the line shape of the optical transitions above the surface-state gap are also found. PMID- 15783932 TI - Narrow luminescence linewidth of a silicon quantum dot. AB - Single-dot luminescence spectroscopy was used to study the emission linewidth of individual silicon nanocrystals from low temperatures up to room temperature. The results show a continuous line narrowing towards lower temperatures with a linewidth as sharp as 2 meV at 35 K. This value, clearly below the thermal broadening at this temperature, proves the atomiclike emission from silicon quantum dots subject to quantum confinement. The low temperature measurements further reveal a approximately 6 meV replica, whose origin is discussed. In addition, an approximately 60 meV TO-phonon replica was detected, which is only present in a fraction of the dots. PMID- 15783933 TI - Magnetic order and spin dynamics in ferroelectric HoMnO3. AB - Hexagonal HoMnO3 is a frustrated antiferromagnet (T(N)=72 K) ferroelectric (T(C)=875 K) in which these two order parameters are coupled. Our neutron measurements of the spin-wave dispersion for the S=2 Mn3+ on the layered triangular lattice are well described by a two-dimensional nearest-neighbor Heisenberg exchange J=2.44 meV, and an anisotropy D that is 0.28 meV above the spin-reorientation transition at 40 K and 0.38 meV below. For H parallel c the magnetic structures and phase diagram have been determined, and reveal additional transitions below 8 K where the ferroelectrically displaced Ho3+ ions are ordered magnetically. PMID- 15783934 TI - Microstructure and thermodynamics of inhomogeneous polymer blends and solutions. AB - A free energy density functional theory (DFT) for nonuniform polymeric mixtures is proposed based on first order thermodynamic perturbation theory. The segment density based free energy functional provides an accuracy comparable to the numerically intensive polymeric DFTs while preserving the computational simplicity of an atomic DFT. The presented applications for solutions and blends of branched and linear polymers demonstrate the capability of the theory to capture the entropic and enthalpic effects governing the microstructure. PMID- 15783935 TI - Dynamics of fluctuations below a stationary bifurcation to electroconvection in the planar nematic liquid crystal N4. AB - We fitted C(k,tau,epsilon) proportional to exp([-sigma(k,epsilon)tau] to time correlation functions C(k,tau,epsilon) of structure factors S(k,t,epsilon) of shadowgraph images of fluctuations below a supercritical bifurcation at V(0)=V(c) to electroconvection of a planar nematic liquid crystal in the presence of a voltage V=sqrt[2]V(0)cos((2pift) [k=(p,q) is the wave vector and epsilon identical with V(2)(0)/V(2)(c)-1]. There were stationary oblique (normal) rolls at small (large) f. Fits of a modified Swift-Hohenberg form to sigma(k,epsilon) gave f-dependent critical behavior for the minimum decay rates sigma(0)(epsilon) and the correlation lengths xi(p,q)(epsilon). PMID- 15783936 TI - Hydrodynamic singularities and clustering in a freely cooling inelastic gas. AB - We employ hydrodynamic equations to follow the clustering instability of a freely cooling dilute gas of inelastically colliding spheres into a well-developed nonlinear regime. We simplify the problem by dealing with a one-dimensional coarse-grained flow. We observe that at a late stage of the instability the shear stress becomes negligibly small, and the gas flows solely by inertia. As a result the flow formally develops a finite-time singularity, as the velocity gradient and the gas density diverge at some location. We argue that flow by inertia represents a generic intermediate asymptotic of unstable free cooling of dilute inelastic gases. PMID- 15783937 TI - Precise toppling balance, quenched disorder, and universality for sandpiles. AB - A single sandpile model with quenched random toppling matrices captures the crucial features of different models of self-organized criticality. With symmetric matrices avalanche statistics falls in the multiscaling Bak-Tang Wiesenfeld universality class. In the asymmetric case the simple scaling of the Manna model is observed. The presence or absence of a precise toppling balance between the amount of sand released by a toppling site and the total quantity the same site receives when all its neighbors topple once determines the appropriate universality class. PMID- 15783938 TI - Scaling behavior of laser population dynamics with time-delayed coupling: theory and experiment. AB - We study the influence of asymmetric coupling strengths on the onset of light intensity oscillations in an experimental system consisting of two semiconductor lasers cross coupled optoelectronically with a time delay. We discover a scaling law that relates the amplitudes of oscillations and the coupling strengths. These observations are in agreement with a theoretical model. These results could be applicable to the population dynamics of other systems, such as the spread of disease in human populations coupled by migration. PMID- 15783939 TI - Passive or active fluctuations in membranes containing proteins. AB - We have experimentally investigated the effect of a transmembrane protein, the Ca2+-ATPase, on shape fluctuations of giant vesicles. By using the micropipette method, we have measured a substantial renormalization of the bending modulus due to the presence of proteins in the membrane. Moreover, we have produced the first quantitative measurement of the active force dipole associated with the amplification of the fluctuations when the proteins are activated by adenosine 5' triphosphate (ATP). PMID- 15783940 TI - Membrane potential and response properties of populations of cortical neurons in the high conductance state. AB - Because of intense synaptic activity, cortical neurons are in a high conductance state. We show that this state has important consequences on the properties of a population of independent model neurons with conductance-based synapses. Using an adiabaticlike approximation we study both the membrane potential and the firing probability distributions across the population. We find that the latter is bimodal in such a way that at any particular moment some neurons are inactive while others are active. The population rate and the response variability are also characterized. PMID- 15783941 TI - Number and length of attractors in a critical Kauffman model with connectivity one. AB - The Kauffman model describes a system of randomly connected nodes with dynamics based on Boolean update functions. Though it is a simple model, it exhibits very complex behavior for "critical" parameter values at the boundary between a frozen and a disordered phase, and is therefore used for studies of real network problems. We prove here that the mean number and mean length of attractors in critical random Boolean networks with connectivity one both increase faster than any power law with network size. We derive these results by generating the networks through a growth process and by calculating lower bounds. PMID- 15783942 TI - Comment on "Thermoinduced magnetization in nanoparticles of antiferromagnetic materials". PMID- 15783944 TI - Efficient universal programmable quantum measurements. AB - A universal programmable detector is a device that can be tuned to perform any desired measurement on a given quantum system, by changing the state of an ancilla. With a finite dimension d for the ancilla only approximate universal programmability is possible, with size d=f(epsilon(-1)) increasing the function of the "accuracy" epsilon(-1). In this Letter we show that, much better than the exponential size known in the literature, one can achieve polynomial size. An explicit example with linear size is also presented. Finally, we show that for covariant measurements exact programmability is feasible. PMID- 15783945 TI - Fermion superfluids of nonzero orbital angular momentum near resonance. AB - We study the pairing of Fermi gases near the scattering resonance of the l not equal 0 partial wave. Using a model potential which reproduces the actual two body low energy scattering amplitude, we have obtained an analytic solution of the gap equation. We show that the ground state of l=1 and l=3 superfluids are orbital ferromagnets with pairing wave functions Y11 and Y32, respectively. For l=2, there is a degeneracy between Y22 and a "cyclic state." Dipole energy will orient the angular momentum axis. The gap function can be determined by the angular dependence of the momentum distribution of the fermions. PMID- 15783946 TI - Laser cooling of fermions to the superfluid transition temperature. AB - Numerical simulations for realistic experimental parameters demonstrate that laser cooling on the attractive side of the Feshbach resonance can drive fermions much below the superfluid transition. For the assumed set of experimental parameters the transition takes place at 0.35T(F), and laser cooling can drive the system down to at least 0.085T(F) in a time of a few seconds. Superfluid growth is self-consistently included in simulations. PMID- 15783947 TI - Symmetry breaking and enhanced condensate fraction in a matter-wave bright soliton. AB - An exact diagonalization study reveals that a matter-wave bright soliton and the Goldstone mode are simultaneously created in a quasi-one-dimensional attractive Bose-Einstein condensate by superpositions of quasidegenerate low-lying many-body states. Upon formation of the soliton the maximum eigenvalue of the single particle density matrix increases dramatically, indicating that a fragmented condensate converts into a single condensate as a consequence of the breaking of translation symmetry. PMID- 15783948 TI - Atom Michelson interferometer on a chip using a Bose-Einstein condensate. AB - An atom Michelson interferometer is implemented on an "atom chip." The chip uses lithographically patterned conductors and external magnetic fields to produce and guide a Bose-Einstein condensate. Splitting, reflecting, and recombining of condensate atoms are achieved by a standing-wave light field having a wave vector aligned along the atom waveguide. A differential phase shift between the two arms of the interferometer is introduced by either a magnetic-field gradient or with an initial condensate velocity. Interference contrast is still observable at 20% with an atom propagation time of 10 ms. PMID- 15783949 TI - Physical bounds to the entropy-depolarization relation in random light scattering. AB - We present a theoretical study of multimode scattering of light by optically random media, using the Mueller-Stokes formalism which permits us to encode all the polarization properties of the scattering medium in a real 4 x 4 matrix. From this matrix two relevant parameters can be extracted: the depolarizing power D(M) and the polarization entropy E(M) of the scattering medium. By studying the relation between E(M) and D(M), we find that all scattering media must satisfy some universal constraints. These constraints apply to both classical and quantum scattering processes. The results obtained here may be especially relevant for quantum communication applications, where depolarization is synonymous with decoherence. PMID- 15783950 TI - Spectroscopy on two coupled superconducting flux qubits. AB - We have performed spectroscopy measurements on two coupled flux qubits. The qubits are coupled inductively, which results in a sigma(z)(1)sigma(z)(2) interaction. By applying microwave radiation, we observe resonances due to transitions from the ground state to the first two excited states. From the position of these resonances as a function of the applied magnetic field, we observe the coupling of the qubits. The coupling strength agrees with calculations of the mutual inductance. PMID- 15783951 TI - Multiphoton path entanglement by nonlocal bunching. AB - Multiphoton path entanglement is created without applying postselection, by manipulating the state of stimulated parametric down-conversion. A specific measurement on one of the two output spatial modes leads to the nonlocal bunching of the photons of the other mode, forming the desired multiphoton path entangled state. We present experimental results for the case of a heralded two-photon path entangled state and show how to extend this scheme to higher photon numbers. PMID- 15783953 TI - Direct measurements of Ab and Ac using vertex and kaon charge tags at the SLAC detector. AB - Exploiting the manipulation of the SLAC Linear Collider electron-beam polarization, we present precise direct measurements of the parity-violation parameters A(c) and A(b) in the Z-boson-c-quark and Z-boson-b-quark coupling. Quark-antiquark discrimination is accomplished via a unique algorithm that takes advantage of the precise SLAC Large Detector charge coupled device vertex detector, employing the net charge of displaced vertices as well as the charge of kaons that emanate from those vertices. From the 1996-1998 sample of 400 000 Z decays, produced with an average beam polarization of 73.4%, we find A(c)=0.673+/ 0.029(stat)+/-0.023(syst) and A(b)=0.919+/-0.018(stat)+/-0.017(syst). PMID- 15783952 TI - Study of the suppressed decays B- -->[K+pi-](D)K- and B- -->[K+pi-]Dpi-. AB - We report a study of the suppressed decays B--->[K(+)pi(-)](D)K- and B-- >[K(+)pi(-)](D)pi(-), where [K(+)pi(-)](D) indicates that the K+pi(-) pair originates from a neutral D meson. These decay modes are sensitive to the unitarity triangle angle varphi(3). We use a data sample containing 275 x 10(6) BB pairs recorded at the Upsilon(4S) resonance with the Belle detector at the KEKB asymmetric e(+)e(-) storage ring. The signal for B--->[K(+)pi(-)](D)K- is not statistically significant, and we set a limit r(B)<0.27 at 90% confidence level, where r(B) is the magnitude of the ratio of amplitudes |A(B--->D 0K-)/A(B- ->D0K-)|. We observe a signal with 6.4sigma statistical significance in the related mode, B--->[K(+)pi(-)](D)pi(-). PMID- 15783956 TI - Suppressing proton decay in the minimal SO(10) model. AB - We show that in a class of minimal supersymmetric SO(10) models which have been found to be quite successful in predicting neutrino mixings, all proton decay modes can be suppressed by a particular choice of Yukawa textures. The required texture not only fits all lepton and quark masses as well as Cabibbo-Kobayashi Maskawa parameters, but it also predicts neutrino mixing parameter U(e3) and Dirac CP phase sin(/delta(MNS)/ to be 0.07-0.09 and 0.3-0.7, respectively. PMID- 15783957 TI - Proton-neutron interactions and the new atomic masses. AB - Double differences of masses can be used to isolate specific nucleonic interactions. With the new 2003 mass tabulation a significant increase in the number of empirical average proton-neutron interactions of the last nucleons can be extracted. It is shown that they exhibit dramatic and distinctive patterns, especially near doubly magic nuclei, that these patterns can be interpreted with a simple ansatz based on overlaps of proton and neutron orbits, and that the trends in p-n interactions across entire shells can be understood if they are correlated with the fractional shell filling. It is shown how these empirical interactions can be sensitive to changes in shell structure in exotic nuclei. Finally, these results are used to suggest criteria for future mass measurements with new exotic beam facilities. PMID- 15783958 TI - New limits on fundamental weak-interaction parameters from superallowed beta decay. AB - A new survey of all world data on superallowed beta decays provides demanding tests of, and tight constraints on, the weak interaction. In confirmation of the conserved vector current hypothesis, the vector coupling constant G(V) is demonstrated to be constant to better than three parts in 10(4), and any induced scalar current is limited to f(S)/=10(14) W/cm(2)) purely classical correlation is strong enough to account for all of the main features observed in experiments to date. PMID- 15783963 TI - Experimental observation of nanojets formed by heating PbO-coated Pb clusters. AB - We are reporting the first experimental observation of nanojets formed by heating PbO-coated Pb clusters, which has been predicted theoretically by Moseler and Landman. During heating, the hot liquid is ejected through the broken orifice into a vacuum and forms a condensed trail in the shape of a tadpole, as shown in the transmission electron micrographs. The temperature-variable Raman spectra indicate that nanojet formation is closely related to the heating temperature and thus essentially to the pressure in the coated clusters. The pressure inside the shell rises from the inner core's melting and its confined volume expansion. It then drops after the final explosion, dominating the whole nanojetting process. PMID- 15783964 TI - Superfluidlike motion of vortices in light condensates. AB - We demonstrate, through numerical simulations, the generation of stable vortex lattices in light condensates. This can be achieved by propagating several concentric laser beams with nested vortices of different topological charges in an optical material with a cubic-quintic nonlinearity. We have considered several initial conditions, and in all the cases the net topological charges of the resulting lattice is equal to the topological charge of the initial outer vortex. The lattice exhibits rotation similar to vortex motion in superfluids. These vortex arrays could be used to implement all-optical photonic crystal fibers. Our results also apply to Bose-Einstein condensates in the presence of three-body elastic interactions. PMID- 15783965 TI - Resonant optical interactions with molecules confined in photonic band-gap fibers. AB - We investigate resonant nonlinear optical interactions and demonstrate induced transparency in acetylene molecules in a hollow-core photonic-band-gap fiber at 1.5 mum. The induced spectral transmission window is used to demonstrate slow light effects, and we show that the observed broadening of the spectral features is due to collisions of the molecules with the inner walls of the fiber core. Our results illustrate that such fibers can be used to facilitate strong coherent light-matter interactions even when the optical response of the individual molecules is weak. PMID- 15783967 TI - Unified tensile fracture criterion. AB - We find that the classical failure criteria, i.e., maximum normal stress criterion, Tresca criterion, Mohr-Coulomb criterion, and von Mises criterion, cannot satisfactorily explain the tensile fracture behavior of the bulk metallic glass (BMG) materials. For a better description, we propose an ellipse criterion as a new failure criterion to unify the four classical criteria above and apply it to exemplarily describe the tensile fracture behavior of BMGs as well as a variety of other materials. It is suggested that each of the classical failure criteria can be unified by the present ellipse criterion depending on the difference of the ratio alpha=tau(0)/sigma(0). PMID- 15783968 TI - Heavy flags undergo spontaneous oscillations in flowing water. AB - By immersing a compliant yet self-supporting sheet into flowing water, we study a heavy, streamlined, and elastic body interacting with a fluid. We find that above a critical flow velocity a sheet aligned with the flow begins to flap with a Strouhal frequency consistent with animal locomotion. This transition is subcritical. Our results agree qualitatively with a simple fluid dynamical model that predicts linear instability at a critical flow speed. Both experiment and theory emphasize the importance of body inertia in overcoming the stabilizing effects of finite rigidity and fluid drag. PMID- 15783969 TI - Exact equation for curved stationary flames with arbitrary gas expansion. AB - An exact equation describing freely propagating stationary flames with arbitrary values of the gas expansion coefficient is obtained. This equation respects all conservation laws at the flame front, and provides a consistent nonperturbative account of the effect of vorticity produced by the curved flame on the front structure. It is verified that the new equation is in agreement with the approximate equations derived previously in the case of weak gas expansion. PMID- 15783966 TI - Excitable optical waves in semiconductor microcavities. AB - We demonstrate experimentally and theoretically the existence of excitable optical waves in semiconductor microcavities. Although similar to those observed in biological and chemical systems, these excitable optical waves are self confined. This is due to a new dynamical scenario, where a stationary Turning pattern controls the propagation of waves in an excitable medium, thus bringing together the two paradigms of dynamical behavior (waves and patterns) in active media. PMID- 15783970 TI - Production of energetic electrons during magnetic reconnection. AB - The production of energetic electrons during magnetic reconnection is explored with full particle simulations and analytic analysis. Density cavities generated along separatrices bounding growing magnetic islands support parallel electric fields that act as plasma accelerators. Electrons because of their low mass are fast enough to make multiple passes through these acceleration cavities and are therefore capable of reaching relativistic energies. PMID- 15783954 TI - Search for Wbb and WH production in pp collisions at square root [s]=1.96 TeV. AB - We present a search for Wbb production in pp collisions at sqrt[s]=1.96 TeV in events containing one electron, an imbalance in transverse momentum, and two b tagged jets. Using 174 pb(-1) of integrated luminosity accumulated by the D0 experiment at the Fermilab Tevatron collider, and the standard-model description of such events, we set a 95% C.L. upper limit on Wbb production of 6.6 pb for b quarks with transverse momenta p(b)(T)>20 GeV and bb separation in pseudorapidity azimuth space DeltaR(bb)>0.75. Restricting the search to optimized bb mass intervals provides upper limits on WH production of 9.0-12.2 pb for Higgs-boson masses of 105-135 GeV. PMID- 15783971 TI - The saturn target for polar direct drive on the national ignition facility. AB - A new target design concept is proposed for direct-drive implosions while the National Ignition Facility is in its initial, indirect-drive configuration. It differs from earlier "polar-direct-drive" designs by adding a low-Z ring around the capsule equator. Refraction in the plasma formed around this ring permits time-dependent tuning of the capsule drive uniformity. An optimized simulation shows an implosion-velocity nonuniformity at the end of the laser pulse of approximately 1% rms for a cryogenic deuterium-tritium shell, enhancing the prospects for an early direct-drive ignition demonstration on the National Ignition Facility. PMID- 15783972 TI - Emission of electromagnetic pulses from laser wakefields through linear mode conversion. AB - Powerful coherent emission around the plasma oscillation frequency can be produced from a laser wakefield through linear mode conversion. This occurs when the laser pulse is incident obliquely to the density gradient of inhomogeneous plasmas. The emission spectrum and conversion efficiency are obtained analytically, which are in agreement with particle-in-cell simulations. The emission can be tuned to be a radiation source in the terahertz region and with field strengths as large as a few GV/m, suitable for high-field applications. The emission also provides a simple way to measure the wakefield produced for particle acceleration. PMID- 15783973 TI - Observation of radio-wave-induced red hydroxyl emission at low altitude in the ionosphere. AB - We report the discovery of radio-wave-induced red emission of OH Meinel rotation vibrational bands at 629.79 nm. These are the first measurements of artificial aurora below 100 km. We believe that the 629.79-nm OH emission was due to radio wave focusing by sporadic ionization clouds near 80-85 km altitude, thus giving a technique to visualize the low-altitude sporadic ionization and providing insight into ionospheric interactions at these low altitudes. PMID- 15783974 TI - Experimental investigation of high-mach-number 3D hydrodynamic jets at the national ignition facility. AB - The first hydrodynamic experiments were performed on the National Ignition Facility. A supersonic jet was formed via the interaction of a laser driven shock ( approximately 40 Mbar) with 2D and 3D density perturbations. The temporal evolution of the jet's spatial scales and ejected mass were measured with point projection x-ray radiography. Measurements of the large-scale features and mass are in good agreement with 2D and 3D numerical simulations. These experiments provide quantitative data on the evolution of 3D supersonic jets and provide insight into their 3D behavior. PMID- 15783975 TI - Size dependence of incipient dislocation plasticity in Ni3Al. AB - Molecular dynamics simulations are carried out to study the incipient dislocation plasticity in Ni3Al. Dislocation nucleation is found to occur preferentially at energetic atomic clusters with larger-than-average relative displacements. From the simulated distribution of the atomic relative displacements, a scaling model is proposed to predict the size dependence of the incipient plasticity condition in real-sized specimens. PMID- 15783976 TI - Phase diagram of silicon from atomistic simulations. AB - In this Letter we present a calculation of the temperature-pressure phase diagram of Si in a range of pressures covering from -5 to 20 GPa and temperatures up to the melting point. The phase boundaries and triple points between the diamond, liquid, beta-Sn, and Si34 clathrate phases are reported. We have employed efficient simulation techniques to calculate free energies and to numerically integrate the Clausius-Clapeyron equation, combined with a tight-binding model capable of an accuracy comparable to that of first-principles methods. The resulting phase diagram agrees well with the available experimental data. PMID- 15783977 TI - Thermal diffusion shock waves. AB - The Ludwig-Soret effect or thermal diffusion, which refers to the separation of liquid mixtures in a temperature gradient, is governed by a nonlinear, partial differential equation in space and time. It is shown here that the solution to the nonlinear differential equation for a binary mixture predicts the existence of shock waves completely analogous to fluid shocks and obeys an expression for the shock velocity that is an exact analogue of the Rankine-Hugoniot relations. Direct measurements of the time dependent, spatial absorption profile of a suspension of nanometer sized particles subjected to a sinusoidal temperature field generated by a pair of continuous laser beams, as well as self-diffraction experiments, show motion of the particles in agreement with the predictions of nonlinear theory. PMID- 15783978 TI - Step structures on III-V phosphide (001) surfaces: how do steps and Sb affect CuPt ordering of GaInP2? AB - The observation of III-V phosphide (001)-(2 x 2) surfaces makes it possible to solve a long standing mystery of step structures. First-principles calculations show that a bulklike type-B step on a hydrogenated 2 x 2 surface is more stable than a rebonded one by 1.1 eV/unit step. In contrast, this energy difference for a H-free beta(2 x 4) surface is only 0.5 eV/unit step. The large difference explains why the CuPt ordering of GaInP is stronger in metal-organic chemical vapor deposition than in molecular beam epitaxy. However, a minute amount of Sb will preferentially attach to the 2 x 2 surface steps and induce additional step structures that cause ordering disruption. PMID- 15783979 TI - Nucleation and submonolayer growth of C60 on KBr. AB - Noncontact atomic force microscopy has been applied to the prototypical molecule insulator system C60 on KBr to study nucleation and submonolayer growth. Overview images reveal an island growth mode with unusual branching structures. Simultaneous molecular and atomic resolution on the C60 and KBr surfaces, respectively, was obtained revealing a coincident 8x3 superstructure. Also, a 21+/-3 pm apparent height difference was observed in atomic force microscopy topographies between some first layer molecules. One of the initial nucleation sites of the C60 islands was determined by observation of loosely bound molecules at kink sites in monatomic KBr steps, in conjunction with the observation that islands form preferentially at step edges. PMID- 15783955 TI - First measurements of inclusive W and Z cross sections from run II of the fermilab tevatron collider. AB - We report the first measurements of inclusive W and Z cross sections times leptonic branching ratios for pp collisions at square root[s]=1.96 TeV, based on their decays to electrons and muons. The data correspond to an integrated luminosity of 72 pb(-1) recorded with the CDF detector at the Fermilab Tevatron. We test e-mu universality in W decays, and we measure the ratio of leptonic W and Z rates from which the leptonic branching fraction B(W-->lnu) can be extracted as well as an indirect value for the total width of the W and the Cabibbo-Kobayashi Maskawa matrix element, |V(cs)|. PMID- 15783980 TI - Covariance for cone and wedge complete filling. AB - Interfacial phenomena associated with fluid adsorption in two dimensional systems have recently been shown to exhibit hidden symmetries, or covariances, which precisely relate local adsorption properties in different confining geometries. We show that covariance also occurs in three-dimensional systems and is likely to be verifiable experimentally and in Ising model simulations studies. Specifically, we study complete wetting in wedge (W) and cone (C) geometries as bulk coexistence is approached and show that the equilibrium midpoint heights satisfy l(c)(h,alpha)=l(w)(h / 2,alpha), where h measures the partial pressure and alpha is the tilt angle. This covariance is valid for both short-ranged and long-ranged intermolecular forces and identifies both leading and next-to-leading order critical exponents and amplitudes in the confining geometries. PMID- 15783981 TI - Vortex sublattice melting in a two-component superconductor. AB - We consider the vortices in a superconductor with two individually conserved condensates in a finite magnetic field. The ground state is a lattice of cocentered vortices in both order parameters. We find two phase transitions: (i) a "vortex sublattice melting" transition where vortices in the field with lowest phase stiffness ("light vortices") lose cocentricity with the vortices with large phase stiffness ("heavy vortices"), entering a liquid state (the structure factor of the light vortices vanishes continuously; this transition is in the 3Dxy universality class); (ii) a first-order melting transition of the lattice of heavy vortices, in a liquid of light vortices. PMID- 15783982 TI - Preserving the half-metallicity at the Heusler alloy Co2MnSi(001) surface: a density functional theory study. AB - We have studied the stability, the electronic, and the magnetic properties of Co2MnSi(001) thin films for 15 different terminations using density functional theory calculations. The phase diagram obtained by ab initio atomistic thermodynamics shows that in practice the MnSi, pure Mn, or pure Si terminated surfaces can be stabilized under suitable conditions. Analyzing the surface band structure, we find that the pure Mn termination, due to its strong surface subsurface coupling, preserves the half-metallicity of the system, while surface states appear for the other terminations. PMID- 15783983 TI - Reversible thermoelectric nanomaterials. AB - Irreversible effects in thermoelectric materials limit their efficiency and economy for applications in power generation and refrigeration. While electron transport is unavoidably irreversible in bulk materials, here we derive conditions under which reversible diffusive electron transport can be achieved in nanostructured thermoelectric materials. We provide a fundamental thermodynamic explanation for why the optimum density of states in a thermoelectric material is a delta function and for why inhomogeneous doping and segmentation improve the thermoelectric figure of merit. PMID- 15783984 TI - Aharonov-Bohm effect as a probe of interaction between magnetic impurities. AB - We study the effects of the RKKY interaction between magnetic impurities on the mesoscopic conductance fluctuations of a metal ring with dilute magnetic impurities. At sufficiently low temperatures and strong magnetic fields, the loss of electron coherence occurs mainly due to the scattering off rare pairs of strongly coupled magnetic impurities. We establish a relation between the dephasing rate and the distribution function of the exchange interaction within such pairs. In the case of the RKKY exchange interaction, this rate exhibits 1/B(2) behavior in strong magnetic fields. We demonstrate that the Aharonov-Bohm conductance oscillations may be used as a probe of the distribution function of the exchange interaction between magnetic impurities in metals. PMID- 15783985 TI - Pure spin current from one-photon absorption of linearly polarized light in noncentrosymmetric semiconductors. AB - We show that one-photon absorption of linearly polarized light should produce pure spin currents in noncentrosymmetric semiconductors, including even bulk GaAs. We present 14x14 k.p model calculations of the effect in GaAs, including strain, and pseudopotential calculations of the effect in wurtzite CdSe. PMID- 15783986 TI - Influence of linker molecules on charge transport through self-assembled single nanoparticle devices. AB - We investigate electrical characteristics of single-electron electrode/nanoisland/electrode devices formed by alkanedithiol assisted self assembly. Contrary to predictions of the orthodox model for double tunnel junction devices, we find a significant ( approximately fivefold) discrepancy in single-electron charging energies determined by Coulomb blockade (CB) voltage thresholds in current-voltage measurements versus those determined by an Arrhenius analysis of conductance in the CB region. The energies do, however, scale with particle sizes, consistent with single-electron charging phenomena. We propose that the discrepancy is caused by a multibarrier junction potential that leads to a voltage divider effect. Temperature and voltage dependent conductance measurements performed outside the blockade region are consistent with this picture. We simulated our data using a suitably modified orthodox model. PMID- 15783987 TI - Spin degree of freedom in the nu=1 bilayer electron system investigated by nuclear spin relaxation. AB - The nuclear-spin-relaxation rate 1/T(1) has been measured in a bilayer electron system at and around total Landau level filling factor nu=1. The measured 1/T(1), which probes electron spin fluctuations, is found to increase gradually from the quantum Hall (QH) state at low fields through a phase transition to the compressible state at high fields. Furthermore, 1/T(1) in the QH state shows a noticeable increase away from nu=1. These results demonstrate that, as opposed to common assumption, the electron spin degree of freedom is not completely frozen either in the QH or the compressible states. PMID- 15783988 TI - Dissociation of vortex stacks into fractional-flux vortices. AB - We discuss the zero field superconducting phase transition in a finite system of magnetically coupled superconducting layers. Transverse screening is modified by the presence of other layers resulting in topological excitations with fractional flux. Vortex stacks trapping a full flux and present at any finite temperature undergo a dissociation transition which corresponds to the depairing of fractional-flux vortices in individual layers. We propose an experiment with a bilayer system allowing us to identify the dissociation of bound vortex molecules. PMID- 15783989 TI - Superconductivity in the americium metal as a function of pressure: probing the Mott transition. AB - High-pressure measurements of the resistivity of americium metal are reported to 27 GPa and down to temperatures of 0.4 K. The unusual dependence of the superconducting temperature (T(c)) on pressure is deduced. The critical field [H(c)(0) extrapolated to T=0] increases dramatically from 0.05 to approximately 1 T as the pressure is increased, suggesting that the type of superconductivity is changing as pressure increases. At pressures of approximately 16 GPa the 5f electrons of Am are changing from localized to itinerant, and the crystal structure also transforms to a complex one. The role of a Mott-type transition in the development of the peak in T(c) above 16 GPa is postulated. PMID- 15783990 TI - Coexistence of ferromagnetism and superconductivity close to a quantum phase transition: the Heisenberg- to Ising-type crossover. AB - A microscopic mean-field theory of the phase coexistence between ferromagnetism and superconductivity in the weakly ferromagnetic itinerant electron system is constructed, while incorporating a realistic mechanism for superconducting pairing due to the exchange of critical spin fluctuations. The self-consistent solution of the resulting equations determines the superconducting transition temperature which is shown to depend strongly on the exchange splitting. The effect of phase crossover from isotropic (Heisenberg-like) to uniaxial (Ising like) spin fluctuations near the quantum phase transition is analyzed and the generic phase diagram is obtained. This scenario is then applied to the case of itinerant ferromagnet ZrZn2, which sheds light on the proposed phase diagram of this compound. A possible explanation of superconductivity in UGe2 is also discussed. PMID- 15783991 TI - 77Se NMR probe of magnetic excitations of the magic angle effect in (TMTSF)2PF6. AB - We report 77Se spin-lattice relaxation rates for (TMTSF)2PF6, carried out in the regime where a set of spectacular transport anomalies known as the "magic angle effects" are observed. In situ resistance measurements (R(zz)) were used to verify the experimental conditions and give precise sample alignment information. We found that the 77Se T-11 exhibits no significant changes as the magnetic-field orientation is rotated through the magic angles, and conclude that there is no evidence for either a single-particle gap or a spin gap. The clearly observed field-induced spin-density wave transition temperature is also, unexpectedly, not enhanced at the magic angles. PMID- 15783992 TI - Theory of incommensurate magnetic correlations across the insulator superconductor transition of underdoped La2-xSrxCuO4. AB - The main feature in the elastic neutron scattering of La2-xSrxCuO4 is the existence of incommensurate peaks with positions that jump from 45 degrees to 0 degrees at 5% doping. We show that the spiral state of the t-t(')-t('')-J model with realistic parameters describes these data perfectly. We explain why in the insulator the peak is at 45 degrees while it switches to 0 degrees precisely at the insulator-metal transition. The calculated positions of the peaks are in agreement with the data in both phases. PMID- 15783993 TI - Universal scaling relations in molecular superconductors. AB - Scaling relations between the superconducting transition temperature T(c), the superfluid stiffness rho(s), and the normal state conductivity sigma(0)(T(c)) are identified within the class of molecular superconductors. These new scaling properties hold as T(c) varies over 2 orders of magnitude for materials with differing dimensionality and contrasting molecular structure and are dramatically different from the equivalent scaling properties observed for cuprate superconductors. These scaling relations place strong constraints on theories for molecular superconductivity. PMID- 15783994 TI - Role of disorder in Mn:GaAs, Cr:GaAs, and Cr:GaN. AB - We present calculations of magnetic exchange interactions and critical temperature T(c) in Ga1-xMnxAs, Ga1-xCrxAs, and Ga1-xCrxN. The local spin-density approximation is combined with a linear-response technique to map the magnetic energy onto a Heisenberg Hamiltonion, but no significant further approximations are made. We show the following: (i) configurational disorder results in large dispersions in the pairwise exchange interactions; (ii) the disorder strongly reduces T(c); (iii) clustering in the magnetic atoms, whose tendency is predicted from total-energy considerations, further reduces T(c), while ordering the dopants on a lattice increases it. With all the factors taken into account, T(c) is reasonably predicted by the local spin-density approximation in Mn:GaAs without the need to invoke compensation by donor impurities. PMID- 15783995 TI - Weak charge-lattice coupling requires reinterpretation of stripes of charge order in La1-xCaxMnO3. AB - Modulations in manganites attributed to stripes of charge/orbital/spin order are thought to result from strong electron-lattice interactions that lock the superlattice and parent lattice periodicities. Surprisingly in La1-xCaxMnO3 (x>0.5,90 K), convergent beam (3.6 nm spot) electron diffraction patterns rule out charge stacking faults and indicate a superlattice with uniform periodicity. Moreover, large area electron diffraction peaks are sharper than simulations with stacking faults. Since the electron-lattice coupling does not lock the two periodicities (to yield stripes) it may be too weak to strongly localize charge. PMID- 15783996 TI - Entanglement in spin chains and lattices with long-range Ising-type interactions. AB - We consider N initially disentangled spins, embedded in a ring or d-dimensional lattice of arbitrary geometry, which interact via some long-range Ising-type interaction. We investigate relations between entanglement properties of the resulting states and the distance dependence of the interaction in the limit N- >infinity. We provide a sufficient condition when bipartite entanglement between blocks of L neighboring spins and the remaining system saturates and determine S(L) analytically for special configurations. We find an unbounded increase of S(L) as well as diverging correlation and entanglement length under certain circumstances. For arbitrarily large N, we can efficiently calculate all quantities associated with reduced density operators of up to ten particles. PMID- 15783997 TI - Stability of high-density one-dimensional excitons in carbon nanotubes under high laser excitation. AB - Through ultrafast pump-probe spectroscopy with intense pump pulses and a wide continuum probe, we show that interband exciton peaks in single-walled carbon nanotubes (SWNTs) are extremely stable under high laser excitations. Estimates of the initial densities of excitons from the excitation conditions, combined with recent theoretical calculations of exciton Bohr radii for SWNTs, suggest that their positions do not change at all even near the Mott density. In addition, we found that the presence of lowest-subband excitons broadens all absorption peaks, including those in the second-subband range, which provides a consistent explanation for the complex spectral dependence of pump-probe signals reported for SWNTs. PMID- 15783998 TI - Gate control of dynamic nuclear polarization in GaAs quantum wells. AB - Gate control of dynamic nuclear polarization under optical orientation is demonstrated in a Schottky-gated n-GaAs/AlGaAs (110) quantum well by time resolved Kerr rotation measurements. Spin relaxation of electrons due to mechanisms other than the hyperfine interaction is effectively suppressed as the donor induced background electron density is reduced from metallic to insulating regimes. Subsequent accumulation of photoexcited electron spins dramatically enhances dynamic nuclear polarization at low magnetic field, allowing us to tune nuclear spin polarization by external gate voltages. PMID- 15783999 TI - Shallow donors in semiconductor nanoparticles: limit of the effective mass approximation. AB - The spatial distribution of the electronic wave function of a shallow, interstitial Li donor in a ZnO semiconductor nanocrystal has been determined in the regime of quantum confinement by using the nuclear spins as probes. Hyperfine interactions as monitored by electron nuclear double resonance spectroscopy quantitatively reveal the transition from semiconductor to molecular properties upon reduction of the size of the nanoparticles. PMID- 15784000 TI - Observation of the transient rotator phase of n-hexadecane in emulsified droplets with time-resolved two-dimensional small- and wide-angle X-ray scattering. AB - Crystallization of n-hexadecane in emulsion droplets was studied using time resolved two-dimensional small- and wide-angle x-ray scattering with differential scanning calorimetry (2D-SAXS-WAXS-in situ DSC) which provides information about both nano- and subnanoscale structural change. n-hexadecane in droplets reproducibly crystallized into the stable triclinic phase via a transient-rotator phase. This is in contrast with previous results that the rotator phase of n hexadecane was observed only occasionally for bulk samples. Thus we confirmed the existence of rotator phase in n-hexadecane, which is important for the study of crystallization of soft materials. We suggest that the rotator phase at the interface of oil and water plays a precursor role for bulk crystallization. This study demonstrates that 2D-SAXS-WAXS-in situ DSC is a powerful tool for the study of a transient phase. PMID- 15784001 TI - Nematics with quenched disorder: pinning out the origin of memory. AB - Memory effects and glassy behavior have been repeatedly observed in disordered nematic liquid crystals but the connection between these effects and the system topology remained unrevealed. We present an analysis of the local and global topology of the nematic ordering in the presence of quenched disorder and we show that nematics with quenched disorder can be mapped into a system of pinned defect lines and that the memory of the system stems from the pinning of these strings. PMID- 15784002 TI - Scaling and universality of critical fluctuations in granular gases. AB - The total energy fluctuations of a low-density granular gas in the homogeneous cooling state near the threshold of the clustering instability are studied by means of molecular dynamics simulations. The relative dispersion of the fluctuations is shown to exhibit a power-law divergent behavior. Moreover, the probability distribution of the fluctuations presents data collapse as the system approaches the instability, for different values of the inelasticity. The function describing the collapse turns out to be the symmetric of the one found in several molecular equilibrium and nonequilibrium systems. PMID- 15784003 TI - Temporally heterogeneous dynamics in granular flows. AB - Granular simulations are used to probe the particle scale dynamics at short, intermediate, and long time scales for gravity-driven, dense granular flows down an inclined plane. On approach to the angle of repose, where motion ceases, the dynamics become intermittent over intermediate times, with strong temporal correlations between particle motions-temporally heterogeneous dynamics. This intermittency is characterized through large-scale structural events whereby the contact network periodically spans the system. A characteristic time scale associated with these processes increases as the stopped state is approached. These features are discussed in the context of the dynamics of supercooled liquids near the glass transition. PMID- 15784004 TI - Inference of a nonlinear stochastic model of the cardiorespiratory interaction. AB - We reconstruct a nonlinear stochastic model of the cardiorespiratory interaction in terms of a set of polynomial basis functions representing the nonlinear force governing system oscillations. The strength and direction of coupling and noise intensity are simultaneously inferred from a univariate blood pressure signal. Our new inference technique does not require extensive global optimization, and it is applicable to a wide range of complex dynamical systems subject to noise. PMID- 15784005 TI - Recombination dramatically speeds up evolution of finite populations. AB - We study the role of recombination, in the form of bacterial transformation, in speeding up Darwinian evolution. This is done by adding a new process to a previously studied Markov model of evolution on a smooth fitness landscape; this new process allows alleles to be exchanged with those in the surrounding medium. Our results, both numerical and analytic, indicate that, for a wide range of intermediate population sizes, recombination dramatically speeds up the rate of evolutionary advance. PMID- 15784006 TI - Optical rheology of biological cells. AB - A step stress deforming suspended cells causes a passive relaxation, due to a transiently cross-linked isotropic actin cortex underlying the cellular membrane. The fluid-to-solid transition occurs at a relaxation time coinciding with unbinding times of actin cross-linking proteins. Elastic contributions from slowly relaxing entangled filaments are negligible. The symmetric geometry of suspended cells ensures minimal statistical variability in their viscoelastic properties in contrast with adherent cells and thus is defining for different cell types. Mechanical stimuli on time scales of minutes trigger active structural responses. PMID- 15784007 TI - Summarizing complexity in high dimensions. AB - High-dimensional, multispectral data on complex physical systems are increasingly common. As the amount of information in data sets increases, the difficulty of effectively utilizing it also increases. For such data, summary information is required for understanding and modeling the underlying dynamics. It is here proposed to use an extension of computational mechanics [C. R. Shalizi and J. P. Crutchfield, J. Stat. Phys. 104, 817 (2001)] to arbitrary spatiotemporal and spectral dimension, for providing such summary information. An example of the use of these tools to identify state evolution in the brain, an archetypal, complex biophysical system, serves as an illustration. PMID- 15784011 TI - The PEG "consult". PMID- 15784009 TI - Interleukin-1beta induces death in chondrocyte-like ATDC5 cells through mitochondrial dysfunction and energy depletion in a reactive nitrogen and oxygen species-dependent manner. AB - IL-1 (interleukin-1) acts as a key mediator of the degeneration of articular cartilage in RA (rheumatoid arthritis) and OA (osteoarthritis),where chondrocyte death is observed. It is still controversial, however, whether IL-1 induces chondrocyte death. In the present study, the viability of mouse chondrocyte-like ATDC5 cells was reduced by the treatment with IL-1beta for 48 h or longer. IL 1beta augmented the expression of the catalytic gp91 subunit of NADPH oxidase, gp91phox, as well as inducible NO synthase in ATDC5 cells. Generation of nitrated guanosine and tyrosine suggested the formation of reactive nitrogen species including ONOO- (peroxynitrite), a reaction product of NO and O2-, in ATDC5 cells and rat primary chondrocytes treated with IL-1beta. Death of ATDC5 cells after IL 1beta treatment was prevented by an NADPH-oxidase inhibitor, AEBSF[4-(2 aminoethyl)benzene-sulphonyl fluoride], an NO synthase inhibitor, L-NAME (NG nitro-L-arginine methyl ester), and a ONOO- scavenger, uric acid. The viability of ATDC5 cells was reduced by the ONOO(-)-generator 3-(4-morpholinyl)sydnonimine hydrochloride, but not by either the NO-donor 1-hydroxy-2-oxo-3-(N-methyl-2 aminopropyl)-3-methyl-1-triazene or S-nitrosoglutathione. Disruption of mitochondrial membrane potential and ATP deprivation were observed in IL-1beta treated ATDC5 cells, both of which were restored by L-NAME, AEBSF or uric acid. On the other hand, no morphological or biochemical signs indicating apoptosis were observed in these cells. These results suggest that the death of chondrocyte like ATDC5 cells was mediated at least in part by mitochondrial dysfunction and energy depletion through ONOO- formation after IL-1beta treatment. PMID- 15784012 TI - Dilation of difficult benign esophageal strictures. PMID- 15784013 TI - Electrocardiogram. PMID- 15784014 TI - Prevalence and clinical spectrum of gastroesophageal reflux: a population-based study in Asan-si, Korea. AB - BACKGROUND AND AIMS: This study estimated the prevalence and clinical spectrum of gastroesophageal reflux disease (GERD) in Asan-si, Korea, as the prevalence is believed to be lower than in Western countries. METHODS: A cross-sectional survey, using a reliable and valid questionnaire, was performed on randomly selected 2,240 Asan-si residents aged between 18 and 69 yr. All respondents were interviewed at their homes or offices by a team of interviewers. RESULTS: Of the 1,902 eligible subjects, 1,417 (78.4%: male 762; female 655) were surveyed. The prevalence of heartburn occurring at least once a month, at least once a week, and at least twice a week was 4.71% (95% confidence interval (CI), 3.6-5.8), 2.0% (95% CI, 1.2-2.7), and 1.3% (95% CI, 0.7-1.9), respectively. The corresponding figures for acid regurgitation were 4.4% (95% CI, 3.3-5.5) and 2.0% (95% CI, 1.3 2.8), respectively. The prevalence of GERD, defined as heartburn and/or acid regurgitation experienced at least weekly, was 3.5% (95% CI, 2.6-4.5). No significant difference was detected between sexes. The prevalence of heartburn was associated with increasing age (p < 0.001). Nineteen percent of our population reported at least one of the atypical symptoms, for instance, chest pain, dysphagia, globus sensation, asthma, bronchitis, pneumonia, or hoarseness. The frequency of frequent GERD among subjects reporting any of the atypical symptoms was 12.6%, which was higher than that of the subjects without atypical symptoms. Patients with typical reflux symptoms were more common among those with atypical symptoms, compared to those without such symptoms (p < 0.001). Using a logistic regression model after adjusting for age and sex, typical reflux symptoms were associated with chest pain (odds ratio (OR), 9.3; 95% CI, 5.9 14.7), dysphagia (OR, 6.4; 95% CI, 2.8-14.7), globus sensation (OR, 3.9; 95% CI, 1.5-9.7), hoarseness (OR, 4.3; 95% CI, 1.4-13.1), asthma (OR, 2.6; 95% CI, 1.4 4.8), and bronchitis (OR, 1.2; 95% CI, 0.6-2.3). CONCLUSION: The prevalence of GERD was 3.5% in this Korean population. Heartburn and acid regurgitation were significantly associated with chest pain, dysphagia, globus sensation, hoarseness, and asthma. PMID- 15784015 TI - Differences in clinical characteristics between patients with endoscopy-negative reflux disease and erosive esophagitis in Japan. AB - OBJECTIVE: Helicobacter pylori infection and atrophic gastritis are inversely related to erosive esophagitis. Whether these factors affect the pathogenesis of endoscopy-negative reflux disease is not clear. We aimed to elucidate the differences in clinical characteristics between endoscopy-negative erosive disease and erosive esophagitis. METHODS: 253 subjects (89 with endoscopy negative reflux disease and 164 with erosive esophagitis) were studied. Gastric atrophy was assessed by measurement of serum pepsinogen. Logistic regression was used to calculate the odds ratio (OR) and 95% confidence intervals (CI) of endoscopy-negative reflux disease compared with erosive esophagitis. RESULTS: Among GERD patients, female gender (OR = 2.27, 95% CI, 1.25-4.10), smoking (OR = 0.45, 95% CI, 0.22-0.91), and the presence of hiatal hernia (OR = 0.30, 95% CI, 0.17-0.56) were significantly associated with endoscopy-negative reflux disease compared with male gender, not smoking, and absence of hiatal hernia, respectively. Body mass index (BMI) was also significantly associated with a decreased OR for endoscopy-negative reflux disease. Although H. pylori infection and gastric atrophy were significantly more common in patients with endoscopy negative reflux disease, these associations did not persist in a multiple adjustment model. After adjustment for gender, BMI, smoking, and hiatal hernia, a decrease in serum pepsinogen I/II ratio was significantly associated with an increased OR for endoscopy-negative reflux disease (p for trend = 0.018). CONCLUSIONS: Female gender, low BMI, not smoking, absence of hiatal hernia, and severity of gastric atrophy were positively associated with endoscopy-negative reflux disease compared with erosive esophagitis among Japanese patients. PMID- 15784016 TI - Gastroesophageal reflux disease in a low-income region in Turkey. AB - OBJECTIVES: Detailed population-based data regarding the prevalence and symptom profile of gastroesophageal reflux disease (GERD) in underdeveloped and developing Caucasian countries are lacking. The aim of this study was to determine the prevalence and clinical spectrum of GERD in a low-income region in Turkey. METHODS: We used a previously validated reflux questionnaire, which was translated into Turkish and culturally adapted. The questionnaire was applied to 630 randomly selected participants greater than 20 yr old living in a population of 8,857 adults, with a low mean income of 75 dollars/person/month. The reliability and reproducibility of the questionnaire were calculated using the kappa statistic (test-retest). Endoscopy and/or 24-h intraesophageal pH monitoring were used to ascertain its validity in identifying patients with reflux. RESULTS: The prevalence of GERD symptoms was 10% for heartburn, 15.6% for regurgitation, and 20% for either symptom experienced at least weekly (95% CI). Heartburn and regurgitation were associated with noncardiac chest pain (37.3%), dysphagia (35.7%), dyspepsia (42.1%), odynophagia (35.7%), globus, hoarseness, cough, hiccup, nausea, vomiting, belching, and NSAID use, but not with body mass index in both frequent and occasional symptom groups. The prevalence of heartburn symptoms, but not regurgitation, increased significantly with age. CONCLUSIONS: The prevalence of GERD in a low-income population in Turkey was similar to that of developed countries, although with a different symptom profile, namely, a lower incidence of heartburn and a higher incidence of regurgitation and dyspepsia. These findings support the contention that there are a large number of patients worldwide in underdeveloped nations with poorly recognized and largely undertreated GERD. PMID- 15784017 TI - The incidence of adenocarcinoma and squamous cell carcinoma of the esophagus: Barrett's esophagus makes a difference. AB - BACKGROUND: Adenocarcinoma limited to the esophagus (ACE) arises in Barrett's esophagus (BE). The incidence of ACE is therefore restricted to this BE subpopulation, whose size is unknown and which is for 95% unidentified. AIMS: To determine the age- and gender-specific incidence rates of ACE, limited to the BE subpopulation, within a defined geographical area and to compare them with those of squamous cell carcinoma of the esophagus (SCC), which can affect the entire population. METHODS: The age- and gender-specific incidence rates for ACE and adenocarcinoma of the cardia (AGC) were calculated after an expert panel classified 87% of all cases of adenocarcinoma of the esophagus reported to the Danish Cancer Registry over a 6-yr period as ACE or AGC. RESULTS: The age specific incidence rates of ACE for males rose from 0.09/10(5) (30-34 yr) to 14.14/10(5) (80-84 yr), falling to 7.2/10(5) (85+ yr), for females from 0.19/10(5) (45-49 yr) to 2.79/10(5) (80-84 yr), falling to 2.43/10(5) (85+ yr) and yielding a gender ratio of 5.9:1; AGC demonstrated a similar pattern and a gender ratio of 4.26:1. However, the incidence rates of SCC continued rising after the age of 80 yr, with a gender ratio of 2.46:1. CONCLUSIONS: The continuing rise in the SCC incidence rates in the elderly demonstrated that the unexpected decline and fall in the incidence rates of ACE over the age of 80 yr did not result from underdiagnosis but were most probably caused by a declining prevalence rate of BE, restricting the elderly BE subpopulation at risk of developing ACE. PMID- 15784018 TI - Dysplasia and risk of further neoplastic progression in a regional Veterans Administration Barrett's cohort. AB - OBJECTIVES: No published data are available on the risk of further neoplastic progression in Barrett's patients stratified by baseline dysplasia status. Our aims were to estimate and compare the risk of progression to high-grade dysplasia or cancer in groups of Barrett's patients stratified by baseline dysplasia status. METHODS: Consecutive Barrett's cases from 1988-2002 were identified via pathology databases in a regional VA health-care system and medical record data were abstracted. The risk of progression to high-grade dysplasia or cancer was measured and compared in cases with versus without low-grade dysplasia within 1 yr of index endoscopy using survival analysis. RESULTS: A total of 575 Barrett's cases had 2,775 patient-years of follow-up. There were 13 incident cases of high grade dysplasia and two of cancer. The crude rate of high-grade dysplasia or cancer was 1 of 78 patient-years for those with baseline dysplasia versus 1 of 278 patient-years for those without (p= 0.001). One case of high-grade dysplasia in each group underwent successful therapy. One incident cancer case underwent successful resection and the other was unresectable. Two cases with high-grade dysplasia later developed cancer, one died postoperatively, the other was unresectable. When these two cases were included (total of four cancers), the crude rate of cancer was 1 of 274 patient-years for those with baseline dysplasia versus 1 of 1,114 patient-years for those without. CONCLUSIONS: In a large cohort study of Barrett's, incident malignancy was uncommon. The rate of progression to high-grade dysplasia or cancer was significantly higher in those with baseline low-grade dysplasia. These data may warrant reevaluation of current Barrett's surveillance strategies. PMID- 15784019 TI - Combination of age and sex improves the ability to predict upper gastrointestinal malignancy in patients with uncomplicated dyspepsia: a prospective multicentre database study. AB - BACKGROUND AND AIMS: Current guidelines recommend endoscopy for dyspeptic patients >45 yr of age and/or with "alarm" symptoms. The management of younger patients with uncomplicated dyspepsia is controversial. The objective of the study was to identify any risk factors predictive of upper gastrointestinal malignancy in patients with uncomplicated dyspepsia and validate their ability in refining indications for endoscopy. METHODS: Prospective database study of consecutive uninvestigated dyspeptic outpatients undergoing endoscopy was performed. A questionnaire including multiple possible prognostic variables was systematically submitted to patients prior to endoscopic examination. Risk factors for upper gastrointestinal malignancy identified were used to derive a prediction rule subsequently validated on an independent population. RESULTS: A total of 5,224 patients with uncomplicated dyspepsia were considered (training sample). Twenty-two (16 males) had malignancy at endoscopy. These patients were about 20 yr older than patients with no malignancy (p < 0.001). The mean age of females with cancer was almost 10 yr higher compared to males (p= 0.08). Such differences in age were confirmed in a split sample of 3,684 patients (p < 0.001 and p < 0.05, respectively). The age cut-offs identified were 35 yr for males and 56 yr for females. CONCLUSIONS: The age threshold for endoscopy should be lowered in males to decrease the risk of missing cancers, and can be safely increased in females without affecting outcomes. In patients with uncomplicated dyspepsia, the combination of age and gender provides a better discriminant power than age alone. PMID- 15784020 TI - Therapeutic potential of duodenal electrical stimulation for obesity: acute effects on gastric emptying and water intake. AB - OBJECTIVES: No satisfactory treatment is available for obesity. Previous animal studies suggested the therapeutic potential of intestinal electrical stimulation for obesity. The aim of this study was to investigate the effects of duodenal electrical stimulation (DES) on gastric emptying and water intake in healthy humans. METHODS: The study was performed in 12 healthy volunteers intubated with a feeding tube in the duodenum under endoscopy. There were three ring electrodes at the end tip of the tube and the two distal electrodes were used for recording and electrical stimulation. On two separate days, each subject underwent a session of DES with various stimulation parameters, a water-intake test with DES or with sham-DES, and a gastric-emptying test with DES or with sham-DES. RESULTS: DES did not induce any noticeable dyspeptic symptoms. The amount of water drunk by the subjects was significantly reduced from 897 +/- 88 ml with sham-DES to 673 +/- 63 ml with DES (p < 0.002). The mean T(50) of gastric emptying was significantly increased from 113.1 +/- 10.0 min with sham-DES to 176.5 +/- 20.8 min with DES state (p < 0.005). The gastric retention at 2 h was increased with DES (42.8 +/- 4.5% vs 61.4 +/- 4.7%; p < 0.02). CONCLUSIONS: DES delays gastric emptying and reduces water intake. It may have a potential application for the treatment of obesity. PMID- 15784021 TI - Endoscopic variceal ligation plus propranolol versus endoscopic variceal ligation alone in primary prophylaxis of variceal bleeding. AB - BACKGROUND AND AIMS: The role of propranolol in addition to EVL in the prevention of first variceal bleed has not been evaluated. This prospective randomized controlled trial compared endoscopic variceal ligation (EVL) with propranolol and EVL alone in the prevention of first variceal bleed among patients with high-risk varices. PATIENTS AND METHODS: One hundred and forty-four consecutive patients with high-risk varices were randomly allocated to EVL plus propranolol (Gr I, n = 72) or EVL alone (Gr II, n = 72). EVL was done at 2-wk interval till obliteration of varices. In Gr I, incremental dosage of propranolol (sufficient to reduce heart rate to 55 beats/min or 25% reduction from baseline) was administered and continued after obliteration of varices. The endpoints of the study were bleeding and death. RESULTS: The two groups of patients had comparable baseline characteristics; follow-up (Gr I: 13.1 +/- 11.5 months, Gr II: 11.2 +/- 9.9 months), number of cirrhotic and noncirrhotic portal hypertension patients [Gr I 64 (88.6%) and 8 (11.4%), Gr II 63 (87.5%) and 9 (12.5%)], and frequency of Child's A (15 vs 18), B (38 vs 35), and C (19 vs 19). The mean daily propranolol dose achieved in Gr I was 95.6 +/- 38.6 mg. Eleven patients had bleeds, 5 in Gr I and 6 in Gr II. All patients bled before the obliteration of varices, the actuarial probability of first bleed at 20 months was 7% in Gr I and 11% in Gr II (p= 0.72). Six patients died in the combination and 8 in EVL group. All deaths in Gr I were due to nonbleed-related causes, while in Gr II, 2 deaths were bleed related, the actuarial probability of death at 20 months was 8% and 15%, respectively (p= 0.37). The probability of bleed-related death was comparable (p= 0.15). At the end of follow-up, 4 patients in Gr I and 11 in Gr II had recurrence of varices (p= 0.03). Side effects on propranolol were seen in 22% patients, in 8% it had to be stopped. There were no serious complications of EVL. CONCLUSIONS: Both EVL plus propranolol and EVL alone are effective in primary prophylaxis of bleed from high-risk varices. Addition of propranolol does not decrease the probability of first bleed or death in patients on EVL. However, the recurrence of varices is lower if propranolol is added to EVL. PMID- 15784022 TI - Primary prophylaxis against variceal bleeding: beta-blockers, endoscopic ligation, or both? AB - Variceal bleeding is one of the complications of cirrhosis that leads to significant morbidity and mortality. It is recommended that all patients with cirrhosis be screened for gastroesophageal varices and those with large varices should be offered primary prophylaxis. Nonselective beta-blockers (nadolol or propranolol) are the treatment of choice for primary prophylaxis but there are a number of limitations to their use. A number of studies have evaluated the efficacy of variceal band ligation (VBL) in providing primary prophylaxis, either in comparison to no treatment or to beta-blockers. VBL is very effective in preventing the initial bleed when compared to no treatment, but it is not superior to beta-blockers. In this issue of the journal the effect of beta blockers on bleeding in patients undergoing VBL is examined and no benefit compared to VBL alone is shown. Thus, patients with large varices should be treated with beta-blockers and VBL should be offered to those cirrhotics who are unable to take beta-blockers. Further study is required to determine if VBL in combination with beta-blockers is more effective than the beta-blockers alone. PMID- 15784023 TI - Assessment of publication bias in the reporting of EUS performance in staging rectal cancer. AB - INTRODUCTION: Endoscopic ultrasound (EUS) has emerged as a promising diagnostic modality for locoregional staging of rectal cancer. However, as with any new technology, publication bias, the selective reporting of studies featuring positive results, may result in overestimation of the capability of EUS. The aim of this study was to systematically assess for publication bias in the reporting of the accuracy of EUS in staging rectal cancer. METHODS: A MEDLINE search for all published estimates of EUS accuracy in staging rectal cancer between 1985 and 2003 was performed. All retrieved studies were fully published in the English literature. Published studies were analyzed and the following information was abstracted: accuracy of EUS, year of publication, number of subjects studied, impact factor of journal, and type of journal (gastroenterology, surgery, radiology, other). RESULTS: Two hundred and two abstracts were reviewed; 41 publications met the stated criteria for inclusion. EUS T-staging accuracy was reported in 40 studies while EUS N-staging accuracy was reported in 27 studies. The experience of 4, 118 subjects was reported with an overall mean T-staging accuracy of 85.2% (median, 87.5%) and N-staging accuracy of 75.0% (median, 76.0%). There was a paucity of smaller studies expressing low EUS accuracy rates. Both T-staging and N-staging accuracy rates also declined over time with the lowest rates reported in more recent literature. CONCLUSION: The performance of EUS in staging rectal cancer may be overestimated in the literature due to publication bias. This inflated estimate of the capability of EUS may lead to unrealistic expectations of this technology. PMID- 15784024 TI - Insertability and safety of a shape-locking device for colonoscopy. AB - BACKGROUND: Loop formation during colonoscopy insertion results in patient pain and delays in advancement of the colonoscope tip. AIM: To assess the insertability and safety of a novel shape-locking guide (ShapeLock, USGI, San Clemente, CA) that resists looping during colonoscopy. RESULTS: In 54 patients, both a disposable and a partly reusable version of the ShapeLock received high scores (mean scores >9.0 on 1-10 scale) for ease of insertion, and ability to maneuver the colonoscope through the shape locked device. No significant trauma resulted from use of the device. CONCLUSION: A novel shape-locking device for colonoscopy can be readily and safely inserted into the colon and locked. The device warrants controlled evaluation to determine in which, if any cases, it may be useful for either experienced colonoscopists and/or those with less or minimal experience. PMID- 15784025 TI - Sexual transmission of HCV between spouses. AB - BACKGROUND AND AIMS: The sexual transmission of hepatitis C virus (HCV) is debated. By excluding other risk factors, the role of sexual intercourse in the transmission could be detected more accurately. We screened HCV prevalence and risk factors in the spouses of chronic hepatitis C (CHC) patients and followed the seroconversion rate of anti-HCV negative spouses. PATIENTS AND METHODS: Six hundred spouses of CHC patients were recruited. The spouses' HCV risk factors were questioned and the spouses were tested for anti-HCV. The 216 spouses who were anti-HCV negative were checked annually for anti-HCV. RESULTS: Anti-HCV was positive in 12 of 600 (2%) of the spouses. Of the 12 anti-HCV positive spouses, 11 were HCV-RNA positive. Of anti-HCV positive and negative spouse groups, mean age was 52.3 +/- 9.8 and 49.8 +/- 12.4 yr; duration of marriage was 1521 +/- 506.7 and 1532.4 +/- 670.2 wk (p > 0.05); and the number of total sexual intercourse was 434 +/- 295 and 307 +/- 333 (p= 0.055), respectively. In our prospective study, none of the spouses developed anti-HCV seroconversion in mean 35.7 +/- 6.3 months and 257.9 +/- 72.2 sexual intercourse. CONCLUSIONS: Anti-HCV was found positive in 2% of the spouses. None of the seronegative spouses developed seroconversion in the 3-yr follow-up period. This is the first study that stresses the importance of the total number of sexual intercourse in sexual transmission (p= 0.055). Our results of special monogamous group with very limited risk factors support the role of number of total sexual intercourse in HCV transmission. However, the seroprevalence rate of the spouses was still within the upper limit of our country population. PMID- 15784026 TI - Sex and hepatitis C. PMID- 15784027 TI - Strong association between gallstones and cardiovascular disease. AB - BACKGROUND AND AIM: Obesity is closely associated with the increased morbidity and mortality of many common diseases in the Western world, including coronary heart disease (CHD) and gallstone diseases (GD). We have investigated the association between GD and CHD in a cross-sectional study. METHODS AND RESULTS: Subjects who had gallstones visible by ultrasound were considered as cases and subjects negative for gallstones were classified as controls. Positive CHD was defined when the stress test was positive. Body mass index (BMI), waist circumference, blood pressure, serum lipid concentrations, and insulin resistance were measured. The association was estimated by odds ratios using logistic regression models adjusted for confounders. Four hundred and seventy-three subjects (292 males and 181 females) were included, comprising 354 controls and 119 cases. Subjects with GD had higher prevalence of CHD (15.96%) than controls (4.52%) (p < 0.0001). In univariate unconditional logistic regression analysis CHD, BMI >/= 30 kg/m(2), waist circumference, high blood pressure, and HOMA-IR > 2.5 were the most important risk factors for GD. In multivariate analysis (adjusted for age and gender, and BMI) the risk for GD in subjects with CHD was higher (OR 2.84, 95% CI: 1.33-6.07, p < 0.007). CONCLUSIONS: Subjects with CHD have an increased risk to have GD, both diseases are strongly associated and the main characteristics of these subjects are those frequently involved as part of the metabolic syndrome. PMID- 15784028 TI - "Pseudotumoral" hepatic areas in acute alcoholic hepatitis: a computed tomography and histological study. AB - Computed tomography (CT) findings in acute alcoholic hepatitis (AAH) have not been investigated and a "pseudotumoral aspect" of the liver parenchyma has rarely been reported as an exceedingly unusual finding. We observed 11 patients with AAH over a 1-yr period, five of whom underwent CT for concomitant clinical reasons. Arterial enhancement indicated areas of hypervascularized tissue in all cases, and the histological findings were fully consistent with AAH without any sign of malignancy. The present findings suggest that the above pattern is not uncommon in AAH and should be carefully considered in order to avoid diagnostic errors. PMID- 15784029 TI - Duration of hepatic iron exposure increases the risk of significant fibrosis in hereditary hemochromatosis: a new role for magnetic resonance imaging. AB - OBJECTIVES: Hepatic fibrosis is a complication of hereditary hemochromatosis. The aim of this study was to determine whether the product of the magnitude and duration of hepatic iron exposure is related to the risk of significant fibrosis. METHODS: Receiver-operating characteristic curve analysis to determine the utility of hepatic iron concentration (HIC) and age in the diagnosis of low- or high-grade fibrosis was undertaken retrospectively in 60 subjects who had undergone liver biopsy for assessment of hereditary hemochromatosis. A prospective pilot study was then conducted in 10 additional subjects to evaluate utility of magnetic resonance imaging (MRI) measurements of HIC to predict fibrosis. RESULTS: Eighteen subjects had high-grade fibrosis while 42 subjects had low-grade fibrosis. Hepatic iron concentration alone was highly sensitive (100%) but of limited specificity (67%) in diagnosis of high-grade fibrosis. The product of [HIC x age] had a sensitivity and specificity of 100% and 86%, respectively, for diagnosis of high-grade fibrosis. Magnetic resonance imaging measurements also provided accurate assignment of subjects into fibrosis severity groups. CONCLUSIONS: Duration of exposure to iron is important in the development of hepatic fibrosis in hereditary hemochromatosis. The product of HIC and age is highly sensitive and specific for diagnosis of high-grade fibrosis and can be obtained using MRI. PMID- 15784030 TI - Increased acylation stimulating protein concentrations in nonalcoholic fatty liver disease are associated with insulin resistance. AB - OBJECTIVES: As acylation stimulating protein (ASP) acts on adipocytes mainly as a paracrine factor to increase triglyceride synthesis and storage; hypothetically, it may play a similar role in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). METHODS: Forty-six male patients with NAFLD (group A), age matched 30 male patients with chronic viral hepatitis (group B) and 30 age matched and body mass index (BMI)-matched healthy male subjects were enrolled in the study. RESULTS: Among the NAFLD patients, 10 patients (24.4%) had simple steatosis and 36 patients (69.6%) had nonalcoholic steatohepatitis (NASH). The mean levels of ASP, complement 3, insulin, C-peptide, HOMA-IR, triglyceride, and very low-density lipoprotein (VLDL) were significantly higher in group A patients than both controls and group B. ASP levels correlated significantly in a positive manner with BMI, insulin, and HOMA-IR. CONCLUSIONS: Dysregulation of the ASP pathway may have important metabolic consequences in NASH and is associated with insulin resistance. PMID- 15784031 TI - Systemic markers of lipid peroxidation and antioxidants in patients with nonalcoholic Fatty liver disease. AB - OBJECTIVES: The aim of the present study was to examine the systemic parameters of oxidative stress and antioxidants in patients with nonalcoholic fatty liver disease and investigate the relationship between these parameters and clinical and biochemical outcomes. METHODS: Fifty-one male patients with nonalcoholic fatty liver disease (group I), 30 age-matched and body mass index (BMI)-matched healthy male subjects, and 30 age-matched male patients with chronic viral hepatitis (group II) were enrolled in the study. RESULTS: Increased systemic levels of malondialdehyde and depletion of antioxidants such as coenzyme Q10, CuZn-superoxide dismutase, and catalase activity were observed in group I. Coenzyme Q10 and CuZn-superoxide dismutase correlated negatively with increasing necroinflammatory activity and fibrosis. Body fat was negatively associated with plasma coenzyme Q10 levels, while an inverse association was found between plasma catalase levels and TG. However, LDL was positively associated with plasma malondialdehyde levels. CuZn-superoxide dismutase levels were negatively associated with glucose, insulin, and HOMA-IR. In addition, the levels of CuZn superoxide dismutase correlated significantly in a negative manner with BMI. CONCLUSIONS: Our results concerning correlations suggest that disturbances in BMI, body fat, and lipid metabolism may contribute to altered oxidative status in NAFLD, and insulin resistance may be related to decreased antioxidants in NAFLD as well as products of lipid peroxidation. However, although our results suggest interesting correlations, this different mostly "weak" relationships must be taken with caution. PMID- 15784032 TI - Effects of indomethacin on viral replication markers in asymptomatic carriers of hepatitis B: a randomized, placebo-controlled trial. AB - OBJECTIVES: Previous studies have suggested some benefits of nonsteroidal antiinflammatory drugs (NSAIDs) use in patients with chronic viral hepatitis. We evaluated potential effects of indomethacin in asymptomatic carriers of hepatitis B surface antigen (HBsAg). DESIGN: Randomized, placebo-controlled, double-masked clinical trial. METHODS: One hundred and twelve patients who were confirmed to be HBsAg carriers for at least 6 months and had normal liver function tests, normal abdominal sonography, and no sign of cirrhosis were randomly assigned into two groups. One group (56 participants, mean age (+/-SD) 31.7 (+/-9.6) yr, 29 male, mean serum alanine aminotransferase (ALT) (+/-SD) 24.9 (+/-9.2)) received indomethacin capsules (25 mg) three times daily and the other group (56 participants, mean age (+/-SD) 33.8 (+/-10.2) yr, 33 male, mean serum ALT (+/-SD) 24.5 (+/-8.7)) took placebo capsules with identical package and appearance. All participants were under treatment for 6 months and were followed 3 months thereafter. Statistical analyses were performed both by intention-to-treat and on treatment methods. RESULTS: Nine participants in the indomethacin group (16%) and 8 in the placebo group (14%) did not complete the trial. HBsAg seroconversion did not differ by treatment group (2 subjects in each group became seronegative). Hepatitis B virus DNA (HBV-DNA) became negative in sera of 7 participants in the indomethacin group but only in 1 in the placebo group (intention-to-treat p= 0.06; on-treatment p= 0.03). Seroconversion of HBeAg to anti-HBe occurred only in 5 participants in the indomethacin group (intention-to-treat p= 0.06; on treatment p= 0.03). Adverse events included one case of hepatotoxicity and two cases of gastritis in the indomethacin group and one suspected gastritis in the placebo group. CONCLUSIONS: We suggest use of indomethacin only in the subgroup of asymptomatic HBsAg carriers who have detectable HBV-DNA or HBeAg in their sera. PMID- 15784033 TI - Efficacy of short-term interferon therapy for patients infected with hepatitis C virus genotype 2a. AB - BACKGROUND AND AIMS: The efficacy of interferon (IFN)-based antiviral therapy for chronic hepatitis C (CHC) varies depending on predictive factors such as hepatitis C virus (HCV) genotype and viral load. For patients with good predictive factors, a low dose and short course of IFN-based therapy may be adequate. However, there is no evidence about the optimal duration of IFN-based therapy for these patients. The aim of this study was to clarify whether the duration of IFN therapy could be shortened to less than the conventional treatment period for patients with good predictive factors. METHODS: A total of 25 treatment-naive CHC patients with genotype 2a were randomized to receive either IFN monotherapy for 24 wks (group A: long-term IFN therapy, n = 13) or for 6 wks (group B: short-term IFN therapy, n = 12). Patients were monitored for HCV RNA and routine liver function tests during and following treatment, and data were examined according to intention-to-treat analysis. RESULTS: Eleven of 13 patients in group A and all patients in group B completed IFN therapy according to the original planned schedule. At the end of the treatment, viral clearance occurred in all patients. However, 4 patients in group A and 5 in group B relapsed within 6 months of follow-up. There was no significant difference of sustained response rate between group A (53.8%) and group B (58.3%). Among patients who had HCV viral load of <100 kIU/ml, the sustained response rate was 83.3% (5/6) in group A and 100% (5/5) in group B. CONCLUSIONS: In this study, our results suggest that the duration of IFN therapy can be shortened to less than 24 wks in patients with good predictive factors. Further studies, however, should examine the optimal regimen of IFN therapy based on the backgrounds of patients. PMID- 15784034 TI - External validation of biochemical indices for noninvasive evaluation of liver fibrosis in HCV chronic hepatitis. PMID- 15784035 TI - Lack of association of common cystic fibrosis transmembrane conductance regulator gene mutations with primary sclerosing cholangitis. AB - BACKGROUND: Primary sclerosing cholangitis (PSC) is a chronic progressive cholestatic liver disease of uncertain etiology. However, the histologic features of PSC liver disease can resemble those in cystic fibrosis (CF), an inherited disorder caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. We sought to determine if PSC patients have a higher frequency of common CF alleles than disease controls. METHODS: DNA was extracted from peripheral lymphocytes of patients with end-stage liver disease. Samples were obtained before liver transplantation from 59 PSC patients and from three groups of control patients (20 each with primary biliary cirrhosis, autoimmune hepatitis, or hepatitis C). DNA samples were genotyped for 32 common CF mutations, the intron 8 T tract variants, and the M470V variant. RESULTS: One of 59 PSC patients (1.7%) had the common CF mutation (DeltaF508) in one CFTR gene. Two controls (3.3%) carried a single CF mutation (DeltaF508 in one primary biliary cirrhosis patient; W1282X in one hepatitis C patient). These rates do not differ from expected in the general population. The frequency of CFTR variants (5T and M470V) was also similar between PSC patients and controls. CONCLUSIONS: Despite anatomical similarities between CF liver disease and PSC, we could not confirm that PSC patients carried common CF mutations or common CFTR variants in higher than expected frequencies. These data suggest that CFTR dysfunction does not influence the pathogenesis of PSC. PMID- 15784036 TI - The effects of chronic treatment with octreotide versus octreotide plus midodrine on systemic hemodynamics and renal hemodynamics and function in nonazotemic cirrhotic patients with ascites. AB - OBJECTIVES: The adrenergic agonist midodrine improved circulatory and renal dysfunction when acutely administered in nonazotemic cirrhotic patients with ascites while its combination with octreotide has recently been proposed as an effective treatment of type 1 hepatorenal syndrome (HRS). However, the effects of octreotide on systemic hemodynamics and renal function in cirrhotic patients are controversial. This study evaluated the effects of chronic treatment with octreotide versus octreotide plus midodrine on systemic hemodynamics and renal hemodynamics, and function in nonazotemic cirrhotic patients with ascites. METHODS: Twenty-five patients were studied at baseline and 11 days after administration of subcutaneous octreotide 300 mug, b.i.d. alone (n = 12) or together with oral midodrine 7.5 mg, t.i.d. (n = 13). RESULTS: Octreotide did not improve systemic hemodynamics whereas the addition of midodrine significantly decreased cardiac index (CI) and heart rate (HR), and increased mean arterial pressure (MAP) and systemic vascular resistance (SVR). Octreotide caused a decrease in renal vascular resistance (RVR) and increased renal blood flow (RBF) but significantly reduced glomerular filtration rate. The association of midodrine to octreotide did not modify renal hemodynamics and function as compared to baseline while it caused an almost significant minor increase in RVR and a significant minor decrease in RBF as compared to octreotide alone. Consequently, a significant minor increase in glomerular filtration rate was demonstrated. The plasma values of active renin, aldosterone, and glucagon were significantly reduced in either group. CONCLUSIONS: Octreotide does not improve systemic hemodynamics in nonazotemic cirrhotic patients with ascites while it impairs renal function. On the other hand, the addition of midodrine can ameliorate the hyperdynamic circulation without inducing renal dysfunction in these patients. PMID- 15784037 TI - Homocysteine triggers mucosal microvascular activation in inflammatory bowel disease. AB - OBJECTIVES: Increased homocysteine contributes to the pathophysiology of several chronic inflammatory diseases. Whether homocysteine could participate in mucosal inflammation in inflammatory bowel disease (IBD) has not been explored yet. Our aims were to study the levels of plasma and mucosal homocysteine in IBD patients and to assess whether homocysteine can trigger an inflammatory reaction on human intestinal microvascular endothelial cells (HIMECs). METHODS: Homocysteine was measured in the plasma, mucosal biopsy, and lamina propria mononuclear cell (LPMC) supernatants from normal and IBD subjects. HIMEC were cultured in presence of homocysteine, TNF-alpha, or folic acid, alone or in combination. Expression of vascular cell adhesion molecule 1 (VCAM-1) and intercellular cell adhesion molecule 1 was measured by flow cytometry and monocyte chemoattractant protein-1 (MCP-1) production by ELISA. Phosphorylation of p38 and p42/44 was assessed by immunoblot in HIMEC extracts. T-cell- and monocyte-HIMEC adhesion assays were used to evaluate the impact of homocysteine on leukocyte adhesion to intestinal endothelial cells. RESULTS: Patients with IBD displayed significantly higher homocysteine plasma and mucosal levels than control subjects. IBD-derived LPMC released higher homocysteine than control-derived LPMC. Treatment of HIMEC with homocysteine, and synergistically with the combination of TNF-alpha and homocysteine, triggered HIMEC inflammation, resulting in VCAM-1 up-regulation, MCP-1 production, and p38 phosphorylation. These events lead to an increased capacity of HIMEC to adhere T- and monocyte cells and were blocked by folic acid treatment. CONCLUSIONS: Homocysteine is increased in both the mucosa and plasma of patients with Crohn's disease and ulcerative colitis and contributes to the inflammatory state of the mucosal IBD endothelium. Therefore, homocysteine could play a proinflammatory role in IBD, which can be efficiently targeted by folic acid supplementation. PMID- 15784038 TI - Characterization of the alternating bowel habit subtype in patients with irritable bowel syndrome. AB - BACKGROUND: Due to a wide range of symptom patterns, patients with irritable bowel syndrome (IBS) are often subgrouped by bowel habit. However, the IBS subgroup with alternating bowel habits (IBS-A) has been poorly characterized. OBJECTIVES: (i) To determine a set of bowel habit symptom criteria, which most specifically identifies IBS patients with an alternating bowel habit, (ii) to describe IBS-A bowel symptom patterns, and (iii) to compare clinical characteristics among IBS-A, constipation-predominant (IBS-C), and diarrhea predominant IBS (IBS-D). METHODS: One thousand one hundred and two Rome I positive IBS patients were analyzed. Three sets of potential criteria for IBS-A were developed and compared by multirater Kappa test. Gastrointestinal, psychological, extraintestinal symptoms, and health-related quality of life were compared in IBS-A, IBS-C, and IBS-D using chi(2) test and analysis of variance (ANOVA). RESULTS: Stool consistency was determined to be the most specific criteria for alternating bowel habits. IBS-A patients reported rapid fluctuations in bowel habits with short symptom flares and remissions. There was a greater prevalence of psychological and extraintestinal symptoms in the IBS-A subgroup compared to IBS-C and IBS-D. No differences were seen between bowel habit subtypes in health-related quality of life. CONCLUSIONS: IBS-A patients have rapidly fluctuating symptoms and increased psychological comorbidity, which should be taken into account for clinical practice and clinical trials. PMID- 15784039 TI - Prevalence of and factors associated with fecal incontinence in a large community study of older individuals. AB - OBJECTIVES: In this study, we describe the prevalence of fecal incontinence by race, age, sex, the presence of major chronic conditions of stroke and diabetes, and the use of certain psychoactive medications. METHODS: Study subjects are participants in the Chicago Health and Aging Project, a study of older Chicago residents of a geographically defined area. In the period 1993-1996, interviewers conducted a door-to-door census that identified 6,099 individuals who participated in in-home interviews. The interviews included a wide range of questions regarding demographics, medical history, and medication use. The question used to determine the presence of fecal incontinence was: "In the past few months have you ever lost control of your bowels when you didn't want to?" RESULTS: Fecal incontinence was seen in 585 of 6,099 survey responders yielding an overall prevalence of 9.6%. The prevalence of fecal incontinence was strongly associated with age across all demographic groups. We did not observe significant differences in the prevalence for males and females once we adjusted for age. However, the increase in prevalence with age was significantly greater among Blacks than Whites. The use of psychoactive medications was found to be associated with significantly higher odds of fecal incontinence. Diabetes and stroke were associated with a higher prevalence of fecal incontinence. CONCLUSIONS: These cross-sectional analyses offer promising evidence that this common condition is correlated with the presence of certain conditions (e.g., stroke and diabetes) and use of certain psychoactive medications. PMID- 15784040 TI - The management of complicated diverticulitis and the role of computed tomography. AB - PURPOSE: Acute diverticulitis is a disease with a wide clinical spectrum, ranging from a phlegmon (stage Ia), to localized abscesses (stages Ib and II), to free perforation with purulent (stage III) or feculent peritonitis (stage IV). While there is little debate about the best treatment for mild episodes and/or very severe episodes, uncertainty persists about the optimal management for intermediate stages (Ib and II). The aim of our study was therefore to define the role of computed tomography (CT) and to analyze its impact on the management of acute diverticulitis. METHODS: We retrospectively analyzed 511 patients (296 males, 215 females) admitted for acute diverticulitis between January 1994 and December 2003. Excluded were patients with stoma reversal only, "diverticulitis" mimicked by cancer, or significantly deficient patient records. Patients were analyzed either as a whole or subgrouped according to age (<40 yr, >40 yr). A modified Hinchey classification was used to stage the severity of acute diverticulitis. RESULTS: In 99 patients (19.4%), an abscess was found (74 pericolic, 25 pelvic, median diameter: 4.0 cm). CT-guided drainage was performed in 16 patients, one failure requiring a two-stage operation. Whereas conservative treatment failed in 6.8% in patients without abscess or perforation, 22.2% of patients with an abscess required an urgent resection (68.2%, one-stage, 31.8%, two-stage). Recurrence rates were 13% for mild cases, as compared to 41.2% in patients with a pelvic abscess (stage II) treated conservatively with/without CT guided drainage. Of all surgical cases, resection/primary anastomosis was achieved in 73.6% with perioperative mortality of 1.1% and leak rate was 2.1%. CONCLUSIONS: CT evidence of a diverticular abscess has a prognostic impact as it correlates with a high risk of failure from nonoperative management regardless of the patient's age. After treatment of diverticulitis with CT evidence of an abscess, physicians should strongly consider elective surgery in order to prevent recurrent diverticulitis. PMID- 15784041 TI - Symptoms and quality of life in chronic pancreatitis assessed by structured interview and the EORTC QLQ-C30 and QLQ-PAN26. AB - BACKGROUND AND AIMS: Chronic pancreatitis (CP) produces disabling symptoms and requires major clinical interventions over a number of years. There is consensus that quality-of-life (QoL) assessment should be part of assessing the treatment and outcome of CP. These symptoms and treatments resemble those of pancreatic cancer, for which there are validated QoL assessment instruments. The aim of our study was to assess the appropriateness of using the EORTC QoL assessment system for pancreatic cancer (the EORTC QLQ-C30 and QLQ-PAN26) for patients with CP, and to document important issues that affect QoL in these patients. METHODS: A structured literature review was undertaken to determine current approaches to QoL in pancreatic disease. Sixty-six patients with newly diagnosed or treated CP were asked to complete the EORTC QLQ-C30 and QLQ-PAN26 in four countries (Germany, Italy, South Africa, and United Kingdom). Patients were asked to review the appropriateness of the content and structure of the instruments, during a directed interview. Standard psychometric tests were used to assess the reliability and validity of the instruments. Peer review was undertaken to review findings and adapt the QLQ-PAN26 on the basis of the responses obtained. RESULTS: The literature review highlighted the potential value of the EORTC QLQ-C30 and identified the lack of a CP-specific instrument, which had been appropriately developed. There was overwhelming consensus among experts that the EORTC assessment system appeared suitable for use in CP patients. This was endorsed by all patients. Patients identified additional issues related to guilt about the use of alcohol and the burden of trying to abstain. All but one scale (jaundice) exhibited adequate internal consistency (r > 0.70) Construct validity of the QLQ C30 and QLQ-PAN26 showed strong associations between conceptually related scales (r > 0.6, p < 0.001) and significantly discriminated between patients on the basis of performance status and requirement for opiate analgesia. Significant issues affecting QoL in CP patients, in addition to recognized symptoms of the disease, were fear of future health problems, difficulty sleeping, and fatigue. CONCLUSION: The EORTC QLQ-C30 and QLQ-PAN26 appear to be an appropriate assessment system for CP, with the addition of items to cover guilt about alcohol consumption, and the burden of abstention. Patients' QoL is adversely affected by the fear of future health problems, difficulty sleeping, and fatigue. PMID- 15784042 TI - Dysplasia in Barrett's esophagus: limitations of current management strategies. AB - Dysplasia is a very imperfect biomarker for malignancy in Barrett's esophagus. Invasive cancer has been found in 30-40% of esophagi resected because preoperative endoscopic examinations had shown high-grade dysplasia. Reports on the natural history of this disorder are sometimes contradictory, but suggest that 10-30% of patients with high-grade dysplasia in Barrett's esophagus will develop a demonstrable malignancy within 5 yr of the initial diagnosis. Proposed management strategies for high-grade dysplasia include esophagectomy, endoscopic ablative therapies, endoscopic mucosal resection (EMR), and intensive endoscopic surveillance. Endoscopic ablative therapies and EMR may not be effective if neoplastic cells have invaded the submucosa or disseminated through mucosal lymphatic channels, and a number of studies suggest that the endoscopic therapies usually leave metaplastic or neoplastic epithelium with malignant potential behind. Limited data suggest that intensive endoscopic surveillance might be a reasonable approach for elderly or infirm patients, but some patients managed in this fashion have developed incurable esophageal cancers. The fundamental question of what is the appropriate length of follow-up for studies on dysplasia treatments has not been resolved. Although 5 yr might be considered the absolute minimum duration for a meaningful follow-up on dysplasia therapy, the follow-up duration in most studies is substantially less than 5 yr. Specific recommendations for management based on these considerations are proposed at the end of this report. PMID- 15784043 TI - Efficacy and safety of traditional medical therapies for chronic constipation: systematic review. AB - OBJECTIVES: Constipation is common, and its treatment is unsatisfactory. Although many agents have been tried, there are limited data to support their use. Our aim was to undertake a systematic review of the efficacy and safety of traditional medical therapies for chronic constipation and to make evidence-based recommendations. METHODS: We searched the English literature for drug trials evaluating treatment of constipation by using MEDLINE and PUBMED databases from 1966 to 2003. Only studies that were randomized, conducted on adult subjects, and published as full manuscripts were included. Studies were assigned a quality score based on published methodology. Standard forms were used to abstract data regarding study design, duration, outcome measures, and adverse events. By using the cumulative evidence of published data for each agent, recommendations were made regarding their use following the United States Preventive Services Task Force guidelines. RESULTS: Good evidence (Grade A) was found to support the use of polyethylene glycol (PEG) and tegaserod. Moderate evidence (Grade B) was found to support the use of psyllium, and lactulose. There was a paucity of quality data regarding many commonly used agents including milk of magnesia, senna, bisacodyl, and stool softeners. CONCLUSIONS: There is good evidence to support the use of PEG, tegaserod, lactulose, and psyllium. Surprisingly, there is a paucity of trials for many commonly used agents. These aspects should be considered when designing trials comparing new agents with traditional therapies because their use may not be well validated. PMID- 15784044 TI - Hepatitis B virus replication in damaged endothelial tissues of patients with extrahepatic disease. AB - Hepatitis B virus (HBV) infection may be complicated by extrahepatic manifestations such as polyarteritis nodosa (PAN), glomerulonephritis, polymyositis, and dermatitis, but the etiology of these processes is not yet clear. HBV replication has been demonstrated in a variety of extrahepatic tissues and cell types, but the possible pathogenetic role of extrahepatic HBV replication has not been fully explored in patients with extrahepatic manifestations of HBV infection. In this case series, immunohistochemistry and in situ hybridization studies were performed on extrahepatic tissues from one HBsAg positive patient with PAN and another HBsAg-positive patient with polymyositis, using HBsAg-seronegative control subjects with the same vasculitic disorders as controls. Tissue samples from the two study patients had detectable HBV RNA, replicative intermediates of HBV DNA, as well as HBsAg and HBcAg localized to vascular endothelium. In contrast, HBsAg-negative control patients had no tissue reactivity. Our results suggest that patients with HBV-related extrahepatic disease have evidence of viral replication in damaged extrahepatic endothelial tissues. While further studies would be required to support a hypothesis of causality, these findings suggest a role for both immune complex deposition and viral replication within diseased endothelial tissue in the pathogenesis of these poorly understood extrahepatic disorders. PMID- 15784045 TI - Clinical course of chronic hepatitis C in patients with very high serum alpha fetoprotein levels and normal hepatic imaging. PMID- 15784046 TI - Preventing dysplasia in Barrett's esophagus: are proton pump inhibitors the answer? PMID- 15784047 TI - Panenteric IBD-like disease in a patient with regressive autism shown for the first time by the wireless capsule enteroscopy: another piece in the jigsaw of this gut-brain syndrome? PMID- 15784048 TI - Sorbitol malabsorption: further explanations are needed. PMID- 15784049 TI - Motivational interviewing and the incredible shrinking treatment effect. PMID- 15784050 TI - Alcohol and intimate partner violence: when can we say that heavy drinking is a contributing cause of violence? PMID- 15784051 TI - South Africa: alcohol today. AB - In 1995 Addiction published an editorial on prospects for substance abuse control in South Africa. This paper reflects on where the country is 10 years later. PMID- 15784052 TI - Sibling effects on smoking in adolescence: evidence for social influence from a genetically informative design. AB - AIMS: Behavioral genetic research has suggested that sibling effects on smoking may reflect social rather than genetic processes. We utilize a genetically informative sample of adolescents to test this proposition, focusing on sibling relationship processes (social connectedness) shown to be influential in studies of deviancy. DESIGN: A combined twin-sibling design was employed to disentangle genetic and non-genetic effects. PARTICIPANTS: We utilized a sample of 1421 adolescent sibling pairs participating in the US National Longitudinal Study of Adolescent Health (Add Health). These sibling pairs represent a spectrum of genetic relatedness and include monozygotic twins, dizygotic twins, biological siblings, half-siblings and unrelated siblings. MEASUREMENTS: Participants completed self-report questionnaires on smoking behavior, quality of relationship with their sibling (social connectedness) and peer and parental smoking. FINDINGS: Main effects of both shared environment and genetics were found on adolescent smoking frequency. Social connectedness between siblings moderated shared environmental influences on smoking frequency at each time period, as well as on change in smoking frequency. Shared environmental effects were more pronounced when siblings reported high levels of social connectedness. These environmental sibling effects on smoking were significant after controlling for parent and peer smoking. CONCLUSIONS: This report extends prior research on sibling effects on smoking by identifying specific relationship dynamics that underlie transmission of risk within sibships and providing evidence that such relationship dynamics represent social rather than genetic processes. PMID- 15784053 TI - Brother, can you spare a smoke? Sibling transmission of tobacco use. PMID- 15784054 TI - The value of different methods and models: comment on Slomkowski et al. (2005). PMID- 15784055 TI - Sibling effects on smoking in adolescence: evidence for social influence from a genetically informative design: comment on Slomkowski et al. 2005. PMID- 15784056 TI - The significance of social connectedness: comment on Slomkowski et al. 2005. PMID- 15784059 TI - Reconsidering the evaluation of addiction treatment: from retrospective follow-up to concurrent recovery monitoring. AB - Historically, addiction treatments have been delivered and evaluated under an acute-care format. Fixed amounts or durations of treatment have been provided and their effects evaluated 6-12 months after completion of care. The explicit expectation of treatment has been enduring reductions in substance use, improved personal health and social function, generally referred to as 'recovery'. In contrast, treatments for chronic illnesses such as diabetes, hypertension and asthma have been provided for indeterminate periods and their effects evaluated during the course of those treatments. Here the expectations are for most of the same results, but only during the course of continuing care and monitoring. The many similarities between addiction and mainstream chronic illnesses stand in contrast to the differences in the ways addiction is conceptualized, treated and evaluated. This paper builds upon established methods of during-treatment evaluation developed for the treatment of other chronic illnesses and suggests a parallel evaluation system for out-patient, continuing-care forms of addiction treatment. The suggested system retains traditional patient-level, behavioral outcome measures of recovery, but suggests that these outcomes should be collected and reported immediately and regularly by clinicians at the beginning of addiction treatment sessions, as a way of evaluating recovery progress and making decisions about continuing care. We refer to this paradigm as 'concurrent recovery monitoring' and discuss its potential for producing more timely, efficient, clinically relevant and accountable evaluations. PMID- 15784060 TI - Evaluating explanations of the Australian 'heroin shortage'. AB - AIMS: In this paper we outline and evaluate competing explanations for a heroin shortage that occurred in Australia during 2001 with an abrupt onset at the beginning of 2001. METHODS: We evaluated each of the explanations offered for the shortage against evidence from a variety of sources: government reports, police and drug law enforcement documents and briefings, key informant (KI) interviews, indicator data and research data. RESULTS: No similar shortage occurred at the same time in other markets (e.g. Vancouver, Canada or Hong Kong) whose heroin originated in the same countries as Australia's. The shortage was due most probably to a combination of factors that operated synergistically and sequentially. The heroin market had grown rapidly in the late 1990s, perhaps helped by a decline in drug law enforcement (DLE) in Australia in the early 1990s that facilitated high-level heroin suppliers in Asia to establish large-scale importation heroin networks into Australia. This led to an increase in the availability of heroin, increasingly visible street-based drug markets, increased purity and decreased price of heroin around the country. The Australian heroin market was well established by the late 1990s, but it had a low profit margin with high heroin purity, and a lower price than ever before. The surge in heroin problems led to increased funding of the Australian Federal Police and Customs as part of the National Illicit Drug Strategy in 1998-99, with the result that a number of key individuals and large seizures occurred during 1999-2000, probably increasing the risks of large-scale importation. The combination of low profits and increased success of law enforcement may have reduced the dependability of key suppliers of heroin to Australia at a time when seized heroin was becoming more difficult to replace because of reduced supplies in the Golden Triangle. These factors may have reduced the attractiveness of Australia as a destination for heroin trafficking. CONCLUSIONS: The Australian heroin shortage in 2001 was due probably to a combination of factors that included increased effectiveness of law enforcement efforts to disrupt networks bringing large shipments of heroin from traditional source countries, and decreased capacity or willingness of major traffickers to continue large scale shipments to Australia. PMID- 15784061 TI - Deterioration over time in effect of Motivational Interviewing in reducing drug consumption and related risk among young people. AB - AIM: To test whether beneficial effects of a single session of Motivational Interviewing (MI) on alcohol, tobacco and illicit drug use apparent after 3 months were maintained until 12 months. DESIGN: Cluster randomized trial, allocating 200 young people in the natural groups in which they were recruited to either MI (n = 105) or to an assessment-only control condition (n = 95). SETTING: Ten further education colleges across inner London. PARTICIPANTS: Two hundred young people who were current users of illegal drugs (age range 16-20 years) with whom contact was established through peers trained for the project. INTERVENTION: The intervention was adapted from MI in the form of a topic-based 1-hour single session discussion. MEASUREMENTS: Changes in cigarette, alcohol, cannabis and other drug use and perceptions of risk and harm between the time of recruitment and follow-up interviews after 3 and 12 months. FINDINGS: A satisfactory follow up rate (81%) was achieved. After 12 months, 3-month differences between MI and assessment-only groups have disappeared almost entirely. Unexpected improvements by the assessment-only control group on a number of outcomes suggest the possibility of reactivity to the research assessment at 3-month follow-up. CONCLUSION: In the terms of the original experiment, there is little evidence of enduring intervention effectiveness shown by between-group differences after 12 months. Deterioration of effect is the most probable explanation, although reactivity to 3-month assessment, a late Hawthorne effect, cannot be ruled out. PMID- 15784062 TI - Impacts of federal precursor chemical regulations on methamphetamine arrests. AB - AIMS: The US government regulated precursor chemicals, ephedrine and pseudoephedrine, multiple times to limit methamphetamine production/availability and thus methamphetamine problems. Research has found that the regulations reduced methamphetamine hospital admissions, but authors have argued that other problems were unaffected. This study examines whether the regulations impacted methamphetamine arrests. DESIGN: ARIMA-intervention time-series analysis with control series. SETTING: California (1982-2001). MEASUREMENTS: Dependent variable series: monthly methamphetamine arrests. Control series: monthly marijuana arrests and cocaine/heroin arrests. INTERVENTIONS: Bulk powder ephedrine and pseudoephedrine: regulated November 1989. Products containing ephedrine as the single active medicinal ingredient: regulated August 1995. Pseudoephedrine products: regulated October 1997. Large-scale producers used ephedrine and pseudoephedrine in these forms. Ephedrine combined with other active medicinal ingredients (e.g. various cold medicines)-used mainly by small-scale producers: regulated October 1996. FINDINGS: The regulation targeting small-scale producers (1996) had no significant impact. In contrast, methamphetamine arrests stopped rising and dropped 31% to 45% each of the three times precursor chemicals used by large-scale producers were regulated. Within 3 years of the bulk powder regulation (1989) and again within 2 years of the ephedrine single ingredient regulation (1995), arrests fully rebounded. During the 4 years following the last regulation (pseudoephedrine products, 1997) arrests only partially rebounded. These effects parallel those reported on hospital admissions. The control series were generally unaffected. CONCLUSIONS: Precursor regulations targeting large scale producers impacted methamphetamine arrests, a criminal justice problem, much as they impacted the public health problem of methamphetamine hospital admissions. Ongoing research is needed to determine whether these problems eventually fully rebound from the last regulation. PMID- 15784063 TI - Nefazodone in out-patient treatment of inhaled cocaine dependence: a randomized double-blind placebo-controlled trial. AB - AIMS: To assess the efficacy of oral nefazodone in the treatment of cocaine dependence. DESIGN: A 10-week randomized double-blind clinical trial was performed. METHODS: All 210 subjects fulfilled Diagnostic and Statistical Manual version IV (DSM-IV) criteria for cocaine dependence and were assigned randomly to 300 mg/day of oral nefazodone (N) or placebo (P). Self-reported drug use, retention interval in treatment, adherence to prescription and depressive symptoms were assessed by the Hamilton scale. FINDINGS: Abstinence from cocaine for 3 weeks or more was achieved by 49.5% (N) and 45.7% (P) (P = 0.58), but 16.2% (N) and 22.9% (P) used other drugs during abstinence. The average interval to resumption of drug use was 33.9 days (N) and 36.1 days (P). Adverse effects were reported by 45.8% (N) and 29.5% (P) (P = 0.01). Treatment for these events was needed more often in N (24.0%) than in P (9.5%) (P < 0.02). CONCLUSIONS: These results do not support the indication of nefazodone for out-patient treatment of inhaled cocaine dependence with or without other associated drug dependence diagnoses. PMID- 15784064 TI - The rise of buprenorphine prescribing in England: analysis of NHS regional data, 2001-03. AB - AIMS: Since its launch in the prescribing market in 1999 for the treatment of opiate dependence, buprenorphine has rapidly become established as an alternative to methadone treatment in the United Kingdom. In the absence of evidence of its clinical superiority over methadone, and given its high relative cost, we sought to examine the impact of buprenorphine availability on opiate treatment services in England. METHODS: Quarterly buprenorphine and methadone community prescription figures were obtained for 28 Strategic Health Authorities (SHAs) in England, for the 2-year period September 2001 to September 2003. Rates of buprenorphine prescribing (as proportion of all opiate prescriptions) were examined over time by number of prescriptions and net ingredient cost. RESULTS: Buprenorphine prescription rates increased disproportionately to methadone in all 28 SHAs. By the end of 2003 the number of buprenorphine prescriptions had increased to 23% of all opiate prescriptions, but accounted for 45% of opiate prescription costs in England. Buprenorphine prescribing rates varied substantially across different regions. CONCLUSIONS: Buprenorphine prescribing has increased dramatically and represents a disproportionately large fraction of community opiate prescribing costs. The marked regional variation suggests the need for further research and the development of national guidelines to support rational prescribing and equitable access to treatment. PMID- 15784065 TI - Predicting the early therapeutic alliance in the treatment of drug misuse. AB - AIMS: To predict the early therapeutic alliance from a range of potentially relevant factors, including clients' social relationships, motivation and psychological resources, and counsellors' professional experience and ex-user status. DESIGN: The study recruited 187 clients starting residential rehabilitation treatment for drug misuse in three UK services. Counsellor and client information was assessed at intake, and client and counsellor ratings of the alliance were obtained during weeks 1, 2 and 3. MEASUREMENTS: The intake assessment battery included scales on psychological wellbeing, treatment motivation, coping strategies and attachment style. Client and counsellor versions of the Working Alliance Inventory (WAI-S) were used for weekly alliance measurement. Hierarchical linear models were used to examine the relationship between alliance and predictor variables. FINDINGS: Clients who had better motivation, coping strategies, social support and a secure attachment style were more likely to develop good alliances. Findings with regard to counsellor characteristics were not clear cut: clients rated their relationships with ex user counsellors, experienced counsellors and male counsellors as better, but more experienced counsellors rated their alliances as worse. CONCLUSIONS: The findings offer important leads as to what interventions might improve the therapeutic alliance. Further work will need to establish whether the therapeutic alliance and ultimately treatment outcomes can be enhanced by working on improving clients' motivation and psychosocial resources. PMID- 15784066 TI - Selection and socialization effects of fraternities and sororities on US college student substance use: a multi-cohort national longitudinal study. AB - AIMS: To examine how membership in fraternities and sororities relates to the prevalence and patterns of substance use in a national sample of full-time US college students. DESIGN: Nationally representative probability samples of US high school seniors (modal age 18 years) were followed longitudinally across two follow-up waves during college (modal ages 19/20 and 21/22). SETTING: Data were collected via self-administered questionnaires from US high school seniors and college students. PARTICIPANTS: The longitudinal sample consisted of 10 cohorts (senior years of 1988-97) made up of 5883 full-time undergraduate students, of whom 58% were women and 17% were active members of fraternities or sororities. FINDINGS: Active members of fraternities and sororities had higher levels of heavy episodic drinking, annual marijuana use and current cigarette smoking than non-members at all three waves. Although members of fraternities reported higher levels than non-members of annual illicit drug use other than marijuana, no such differences existed between sorority members and non-members. Heavy episodic drinking and annual marijuana use increased significantly with age among members of fraternities or sororities relative to non-members, but there were no such differential changes for current cigarette use or annual illicit drug use other than marijuana. CONCLUSIONS: The present study provides strong evidence that higher rates of substance use among US college students who join fraternities and sororities predate their college attendance, and that membership in a fraternity or sorority is associated with considerably greater than average increases in heavy episodic drinking and annual marijuana use during college. These findings have important implications for prevention and intervention efforts aimed toward college students, especially members of fraternities and sororities. PMID- 15784067 TI - Gender differences in the association between substance use and elevated depressive symptoms in a general adolescent population. AB - AIMS: This study explores gender differences in the association between substance use and elevated depressive symptoms in the general adolescent population. DESIGN: Cross-sectional self-reported anonymous survey, the 2002/2003 Student Drug Use Survey in the Atlantic Provinces. The sample design was a single-stage cluster sample of randomly selected classes stratified by grade and region. SETTING: The four Atlantic provinces of Canada. PARTICIPANTS: A total of 12 771 students in junior and senior high schools of the public school systems, representing a response rate of about 97%. The average age of participants was 15.2 years. MEASUREMENTS: The measure of elevated depressive symptoms was a 12 item version of the CES-D with three categories of depression risk validated in a companion study. FINDINGS: The prevalence of very elevated depressive symptoms was 8.6% in females and 2.6% in males. Alcohol use and cigarette smoking were found to be independent predictors of elevated depressive symptoms in females, but not males; cannabis use was found to be an independent predictor of elevated depressive symptoms in both males and females. Age was found to have a curvilinear relationship with elevated depressive symptoms in females but not in males. The adolescent's academic performance and province of residence were found to be independent risk factors of elevated depressive symptoms among both males and females. About 10.3% of adolescents considered to be potential candidates for needing help reported having received help because they felt depressed. CONCLUSIONS: The association between depression risk and age, alcohol use, cigarette smoking and cannabis use in the general adolescent population is not straightforward and may differ according to gender. There is unmet need for help for depression among adolescents. PMID- 15784068 TI - Could the high level of cirrhosis in central and eastern Europe be due partly to the quality of alcohol consumed? An exploratory investigation. AB - BACKGROUND: The burden of alcohol-related diseases differs widely among countries. Since the 1980s, a band of countries in Central and Eastern Europe have experienced a steep rise in deaths from chronic liver diseases and cirrhosis. A possible risk factor is the consumption of illegally produced home made spirits in these countries containing varying amounts of aliphatic alcohols and which may be hepatotoxic. However, little is known about the composition of such beverages. AIMS: To compare the concentration of short-chain aliphatic alcohols in spirits from illegal and legal sources in Hungary. DESIGN: Samples taken from commercial retailers and illegal sources were collected and their aliphatic patterns and alcohol concentrations were determined by gas chromatographic/mass spectrometric (GC/MS) analysis. FINDINGS: The concentrations of methanol, isobutanol, 1-propanol, 1-butanol, 2-butanol and isoamyl alcohol were significantly higher in home-made spirits than those of from commercial sources. CONCLUSIONS: The results suggest that the consumption of home-made spirits is an additional risk factor for the development of alcohol-induced cirrhosis and may have contributed to high level of liver cirrhosis mortality in Central and Eastern Europe. Restrictions on supply and sale of alcohol from illicit sources are needed urgently to reduce significantly the mortality from chronic liver disease. PMID- 15784069 TI - Adolescent drinking level and adult binge drinking in a national birth cohort. AB - AIMS: To assess (i) continuities in binge drinking across adulthood and (ii) the association between adolescent drinking level and adult binge drinking. DESIGN: Population-based prospective birth cohort. SETTING: England, Scotland and Wales. PARTICIPANTS: All births during one week in March 1958 (n = 8520 in analysis). MEASUREMENTS: Alcohol consumption reported at 16, 23, 33 and 42 years. Binge drinkers were identified by dividing number of units of alcohol consumed in the last week by usual drinking frequency, with limits of >/=10 units/occasion for men and >/=7 for women. FINDINGS: Four in five cohort members drank alcohol at least twice a month. Prevalences of binge drinking at 23, 33 and 42 years among men were 37%, 28% and 31% and among women 18%, 13% and 14%. Most binge drinkers in adulthood changed drinking status during this period. Nevertheless, binge drinking at age 23 increased the odds of binge drinking at 42 years: odds ratio (OR) 2.10 (95% CI 1.85, 2.39) for men; OR 1.56 (95% CI 1.29,1.89) for women. Women who rarely or never drank aged 16 were less likely than light drinkers (0-2 units/week) to binge drink as adults, OR at 23 years 0.65 (95% CI 0.55, 0.77). Men who were heavier drinkers (>/=7 units/week) at 16 years were more likely than light drinkers to binge drink throughout adulthood; at 42 years, OR 1.64 (95% CI 1.33, 2.08). CONCLUSIONS: Binge drinking is common in British men and women throughout adulthood with continuities between the 20s and 40s. Adolescent drinking has a modest although important association with adult binge drinking. PMID- 15784070 TI - Tobacco abstinence symptom suppression: the role played by the smoking-related stimuli that are delivered by denicotinized cigarettes. AB - AIMS: Cigarette smoking causes cancer and disease, yet people find quitting difficult due to aversive symptoms that accompany tobacco abstinence. Understanding how to suppress these symptoms is critical in developing effective smoking cessation treatments. Pharmacologically, pure nicotine suppresses tobacco abstinence symptoms partially, and non-nicotine, smoking-related stimuli suppress these abstinence symptoms fully, at least for 24 hours. The current study was designed to clarify the impact of smoking-related stimuli on tobacco withdrawal, and to explore the duration of their ability to suppress withdrawal in smokers. DESIGN: Three double-blind, within-subjects, Latin square-ordered, 5-day conditions in which participants smoked nicotinized, denicotinized or no cigarettes. SETTING: Out-patient laboratory at Virginia Commonwealth University. PARTICIPANTS: Thirteen women and 19 men. MEASUREMENTS: Subjective, physiological and performance measures were collected daily and compliance with study conditions was verified objectively. FINDINGS: Smoking-related stimuli are sufficient for suppressing some symptoms of tobacco abstinence over a 5-day period [i.e. Questionnaire of Smoking Urges (QSU) factor 1, 'Desire for sweets', 'Hunger' and 'Urges to smoke'], while in this study a combination of nicotine and smoking-related stimuli suppressed other symptoms (i.e. 'Difficulty concentrating', 'Increased eating', 'Restlessness' and 'Impatient'). CONCLUSIONS: These results indicate that, while some tobacco abstinence symptoms may be suppressed with nicotine, suppressing others may also require strategies that address the absence of smoking-related stimuli. PMID- 15784071 TI - Fatal opiate overdose following regimen changes in naltrexone treatment. PMID- 15784072 TI - Is level of interest among cannabis users in self-help materials and other services aimed at reducing problem use? PMID- 15784073 TI - Reporting bias and self-reported drug use. PMID- 15784080 TI - The role of imaging in urinary incontinence. AB - A detailed history, physical examination, symptom and quality-of-life assessment, and urine analysis are unanimously considered essential components of the initial evaluation of urinary incontinence. Beyond these assessments, there are no universally accepted recommendations and, to date, imaging is not recommended in the initial management of urinary incontinence. In selected patients, urodynamics and/or a radiographic evaluation may be indicated. According to International Continence Society guidelines, imaging of the upper and lower urinary tract is indicated only if renal damage or pelvic pathology are suspected; video urodynamics and voiding cysto-urethrography are considered optional diagnostic tests and continue to be refined; magnetic resonance imaging is considered an important research tool in evaluating lower urinary tract disorders, but at present its clinical role remains investigational. PMID- 15784081 TI - Laparoscopy for impalpable testes. PMID- 15784082 TI - The urological management of the patient with acquired immunodeficiency syndrome. AB - In people infected with the human immunodeficiency virus (HIV) both the CD4 T cell count and the viral load are used to monitor disease progression to acquired immunodeficiency syndrome (AIDS). CD4 counts of <500/mm(3) are associated with opportunistic infections and certain malignancies, so-called 'AIDS-defining' conditions. Highly active antiretroviral therapy, using combinations of reverse transcriptase inhibitors and/or protease inhibitors, can improve considerably the prognosis of people who are HIV-positive, but such therapy is not yet widely available in many developing countries. People with AIDS are predisposed to urinary tract infection (UTI) by uncommon bacteria and pathogens, e.g. fungi, parasites and viruses, which may affect any urogenital organ; treatment should be culture-specific and long-term, because there is a tendency to recurrence, infection with multiple organisms and resistant isolates. Voiding dysfunction in patients with AIDS is usually a result of neurological complications caused by opportunistic infections, and has a poor prognosis. Of patients with AIDS, 30-50% develop a cancer, especially Kaposi's sarcoma (KS) and non-Hodgkin's lymphoma (NHL). KS may involve any urogenital organ, but is usually part of systemic disease. Small lesions on the external genitalia can be treated with laser, cryotherapy or surgical excision, larger lesions with radiotherapy, and disseminated or visceral KS with multidrug chemotherapy. NHL may involve the kidneys, testes and retroperitoneal lymph nodes, thus obstructing the ureters, which may require ureteric stenting or percutaneous nephrostomy. NHL can be treated with radiotherapy and combination chemotherapy. Urolithiasis in patients with AIDS may be caused by indinavir, a protease inhibitor, but the more common types of stones may also occur. Fluid-electrolyte and acid-base disturbances are common in patients with advanced AIDS, secondary to vomiting, diarrhoea, malnutrition or septicaemia. HIV-associated nephropathy occurs in 10-30% of patients, and often leads to renal failure. Testicular atrophy is common, leading to infertility, erectile dysfunction (ED) and decreased libido. Treatment for ED must include counselling about strategies to reduce the transmission of HIV. The risk of HIV transmission after parenteral exposure to blood from an HIV-positive patient is relatively low (0.2-0.4%); the urologist can reduce the risk of transmission during surgery by adopting certain precautions. After occupational exposure to HIV, chemoprophylaxis with antiretroviral medication can significantly reduce the probability of HIV transmission. PMID- 15784083 TI - Clinical trials in urology: how many patients are required to achieve statistically significant results? AB - During the clinical validation of a new drug there are several clinical phases. Once phase II studies have defined the efficacy of a new drug, clinical research is used to evaluate its significance in clinical practice, comparing it with other drugs or treatments in use for similar clinical conditions. The group of patients undergoing standard treatment (either untreated or treated with placebo) is thus used as a control; these phase III studies are termed 'controlled clinical studies'. The general condition for comparing patients treated with the new drug is that they do not have characteristics (relevant to the study) that are systematically different from those in the control group. Randomization guarantees comparability between treated and untreated (or otherwise treated) patients. The comparability of the observations of the studied events are guaranteed by blinding and placebo. The fundamental question when designing a controlled clinical study to evaluate whether there are differences between two or more treatments is how many patients are needed. Generally, the smaller the clinically relevant differences in efficacy between treatments, the more patients are required, to provide sufficient statistical power and meaningful clinical results. A group of randomized patients represents the final point of sequential steps. Also of importance is to what kind of 'population' the results from the studied sample can be applied (qualitatively, not necessarily quantitatively), i.e. the general applicability of a study, or whether the findings can be used to treat future patients with the same or similar characteristics as those randomized, or to all patients with the same pathology. Answers to these questions depend on many aspects of the randomized selection mechanisms, the disease characteristics, and knowledge of the biological effects of the drug to be tested. PMID- 15784084 TI - Need for approval and children's well-being. AB - This research examined the hypothesis that a tendency to base one's self-worth on peer approval is associated with positive and negative aspects of children's well being. A sample of 153 fourth through eighth graders (9.0 to 14.8 years) reported on need for approval, global self-worth, social-evaluative concerns, anxiety and depression, and exposure to victimization. Teachers reported on social behavior. Results confirmed that need for approval is a two-dimensional construct composed of positive (enhanced self-worth in the face of social approval) and negative (diminished self-worth in the face of social disapproval) approval-based self appraisals. Need for approval had trade-offs for well-being that depended on the dimension (positive vs. negative), the psychological domain (emotional vs. social adjustment), children's sex and age, and children's social context (high vs. low peer victimization). PMID- 15784085 TI - Children's implicit learning of graphotactic and morphological regularities. AB - In French, the transcription of the same sound can be guided by both probabilistic graphotactic constraints (e.g., /epsilon t/ is more often transcribed ette after -v than after -f) and morphological constraints (e.g., /epsilon t/ is always transcribed ette when used as a diminutive suffix). Three experiments showed that pseudo-word spellings of 8-to 11-year-old children and adults were influenced by both types of constraints. The influence of graphotactic regularities persisted when reliance on morphological rules was possible, without any falling off as a function of age. This suggests that rules are not abstracted, even after massive amounts of exposure to a rule-based material. These results can be accounted for by a statistical model of implicit learning. PMID- 15784086 TI - Mapping novel nouns and verbs onto dynamic action events: are verb meanings easier to learn than noun meanings for Japanese children? AB - The present research examined how 3- and 5-year-old Japanese children map novel nouns and verbs onto dynamic action events and generalize them to new instances. Studies 1 to 3 demonstrated that although both 3- and 5-year-olds were able to map novel nouns onto novel objects, only 5-year-olds could generalize verbs solely on the basis of the sameness of the action. Study 4 showed that the difficulty young children experience in learning verbs lies mainly in mapping the appropriate element to a verb rather than in encoding and remembering an action itself. The results of this research are related to a long-debated issue of whether noun learning is privileged over verb learning. PMID- 15784087 TI - Origins of individual differences in theory of mind: from nature to nurture? AB - In this study of the origins of individual differences in theory of mind (ToM), the Environmental Risk (E-Risk) Longitudinal Twin Study sample of 1,116 sixty month-old twin pairs completed a comprehensive battery of ToM tasks. Individual differences in ToM were striking and strongly associated with verbal ability. Behavioral genetic models of the data showed that environmental factors explained the majority of the variance in ToM performance in this sample. Shared environmental influences on verbal ability had a common impact on ToM and explained more than half the phenotypic correlation between these two skills. Possible underlying proximal mechanisms are discussed, including maternal speech and mind-mindedness, sibling interactions, and peer influences. PMID- 15784088 TI - Parent-child participation in planning children's activities outside of school in European American and Latino families. AB - This longitudinal research used a sociocultural perspective to examine planning competence in the everyday experiences of European American and Latino children from 7 to 9 years of age. Data on children's participation in planning their activities outside of school, parental expectations about children's planning competence, and children's planning in the classroom were collected yearly from Grades 2 to 4 from 140 children and their mothers, and the children's teachers. Results indicate that decision-making practices and parental expectations change with development and vary by ethnicity. Decision making at home was related to children's classroom planning; however, the nature of these relations changed over middle childhood. Results are discussed in terms of cultural and parental contributions to the development of planning skills. PMID- 15784089 TI - Intergenerational transmission of warm-sensitive-stimulating parenting: a prospective study of mothers and fathers of 3-year-olds. AB - More than 200 New Zealand men and women studied repeatedly since age 3 were videotaped interacting with their own 3-year-old children to determine (a) whether childrearing and family climate experienced in 3 distinct developmental periods while growing up (i.e., early childhood, middle childhood, early adolescence) predicted parenting and (b) whether romantic relationship quality moderated the effect of childrearing history on observed parenting. Support for the first hypothesis emerged across all 3 developmental periods for mothers (only), with no evidence of moderating effects of romantic relationship quality for mothers or fathers. Results are discussed in terms of supportive versus harsh parenting, mother-father differences, and the characteristics of the sample. PMID- 15784090 TI - Academic self-concept, interest, grades, and standardized test scores: reciprocal effects models of causal ordering. AB - Reciprocal effects models of longitudinal data show that academic self-concept is both a cause and an effect of achievement. In this study this model was extended to juxtapose self-concept with academic interest. Based on longitudinal data from 2 nationally representative samples of German 7th-grade students (Study 1: N = 5,649, M age = 13.4; Study 2: N = 2,264, M age = 13.7 years), prior self-concept significantly affected subsequent math interest, school grades, and standardized test scores, whereas prior math interest had only a small effect on subsequent math self-concept. Despite stereotypic gender differences in means, linkages relating these constructs were invariant over gender. These results demonstrate the positive effects of academic self-concept on a variety of academic outcomes and integrate self-concept with the developmental motivation literature. PMID- 15784091 TI - The peer group as a context: moderating effects on relations between maternal parenting and social and school adjustment in chinese children. AB - This 2-year longitudinal study examined, in a sample of Chinese children (initial M age = 11 years), the moderating effects of the peer group on relations between maternal supportive parenting and social and school adjustment. Data were collected from multiple sources including peer assessments, teacher ratings, school records, and maternal reports. It was found that whereas group prosocial cooperative functioning strengthened the role of supportive parenting in helping children develop social and school competence, group antisocial-destructive functioning undermined the contributions of supportive parenting to children's social and academic achievement. The results indicated the significance of the peer group as a social context for socialization and development in Chinese children. PMID- 15784092 TI - Daily reports of witnessing and experiencing peer harassment in middle school. AB - Two studies examined daily incidents of peer harassment in urban middle schools. Sixth-grade students (M age = 11 years) described their daily personal experiences and witnessed accounts of peer harassment, and rated their negative feelings across a 2-week period. In Study 1 (n = 95), within-subject analyses across 4 days revealed that both personally experienced and witnessed harassment were associated with increases in daily anxiety, whereas witnessing harassment buffered students against increases in humiliation on days when they personally experienced harassment. Evidence for witnessing as a buffer against increases in humiliation and anger was also found in Study 2 (n = 97) that included 5 daily reports. Witnessing harassment also protected students against increases in negative self-perceptions. PMID- 15784093 TI - Social norms and self-presentation: children's implicit and explicit intergroup attitudes. AB - Two studies examined whether social norms and children's concern for self presentation affect their intergroup attitudes. Study 1 examined racial intergroup attitudes and normative beliefs among children aged 6 to 16 years (n=155). Accountability (i.e., public self-focus) was experimentally manipulated, and intergroup attitudes were assessed using explicit and implicit measures. Study 2 (n = 134) replicated Study 1, focusing on national intergroup attitudes. Both studies showed that children below 10 years old were externally motivated to inhibit their in-group bias under high public self-focus. Older children were internally motivated to suppress their bias as they showed implicit but not explicit bias. Study 1, in contrast to Study 2, showed that children with low norm internalization suppressed their out-group prejudice under high public self focus. PMID- 15784094 TI - Mothers' time with infant and time in employment as predictors of mother-child relationships and children's early development. AB - This study tested predictions from economic and developmental theories that maternal time with an infant is important for mother-child relationships and children's development, using time-use diaries for mothers of 7- to 8-month-old infants from the National Institute of Child Health and Human Development Study of Early Child Care (N = 1,053). Employment reduced time with infants, but mothers compensated for some work time by decreasing time in other activities. With family and maternal characteristics controlled, time with infants predicted high Home Observation for Measurement of the Environment (HOME) scores and maternal sensitivity, but bore little relation to children's engagement with mothers, secure attachment, social behavior, or cognitive performance from 15 to 36 months. Mothers who spent more time at work had higher HOME scores. Maternal time with infants may reflect maternal characteristics that affect both time allocation and maternal behavior. PMID- 15784095 TI - Examining the motivational impact of intrinsic versus extrinsic goal framing and autonomy-supportive versus internally controlling communication style on early adolescents' academic achievement. AB - The present experimental research examined whether framing early adolescents' (11 to 12-year-olds) learning activity in terms of the attainment of an extrinsic (i.e., physical attractiveness) versus intrinsic (i.e., health) goal and communicating these different goal contents in an internally controlling versus autonomy-supportive way affect performance. Both conceptual and rote learning were assessed. Three experimental field studies, 2 among obese and 1 among nonobese participants, confirmed the hypothesis that extrinsic goal framing and internal control undermine conceptual (but not rote) learning, even in comparison with a control group. Study 3 indicated that the positive effect of intrinsic goal framing on conceptual learning was mediated by task involvement, whereas the positive effect of autonomy-supportive communication style on conceptual learning was mediated by relative autonomous motivation. PMID- 15784096 TI - Steps in theory-of-mind development for children with deafness or autism. AB - Prior research demonstrates that understanding theory of mind (ToM) is seriously and similarly delayed in late-signing deaf children and children with autism. Are these children simply delayed in timing relative to typical children, or do they demonstrate different patterns of development? The current research addressed this question by testing 145 children (ranging from 3 to 13 years) with deafness, autism, or typical development using a ToM scale. Results indicate that all groups followed the same sequence of steps, up to a point, but that children with autism showed an importantly different sequence of understandings (in the later steps of the progression) relative to all other groups. PMID- 15784097 TI - Infant emotional and cortisol responses to goal blockage. AB - This study examined the relation of infant emotional responses of anger and sadness to cortisol response in 2 goal blockage situations. One goal blockage with 4-month-old infants (N = 56) involved a contingency learning procedure where infants' learned response was no longer effective in reinstating an event. The other goal blockage with 6-month-old infants (N = 84) involved the still face procedure where infants' reactions to their mothers' lack of responsivity were not effective in reestablishing interaction. For both blockages, sadness was related to cortisol response, though anger was not--the greater the sadness, the higher the cortisol response. This differential relation is consistent with other evidence indicating the more positive role of anger as opposed to sadness in overcoming an obstacle. PMID- 15784098 TI - H+/K+-ATPase (proton pump) inhibitors dampen increased cough reflex: more than gastric acid suppression. PMID- 15784099 TI - Are specific immunoglobulin E titres reliable for prediction of food allergy? PMID- 15784100 TI - Indirect bronchial hyper-responsiveness: the coming of age of a specific group of bronchial challenges. PMID- 15784101 TI - Attenuating effect of H+K+ATPase inhibitors on airway cough hypersensitivity induced by allergic airway inflammation in guinea-pigs. AB - BACKGROUND: Gastrooesophageal reflux (GER) is a frequent cause of chronic cough. Several investigators have indicated that inhibitors of H(+)K(+)ATPase (proton pump inhibitors; PPIs) could relieve coughing via inhibition of acid reflux. However, we considered that PPIs might directly inhibit increased cough reflex sensitivity. OBJECTIVE: The present study was designed to examine whether PPIs directly inhibit antigen-induced increase in cough reflex sensitivity and to elucidate the mechanism. METHODS: Actively sensitized guinea-pigs were challenged with aerosol antigen (ovalbumin, OVA) and cough reflex sensitivity to inhaled capsaicin was measured 24 h later. The PPIs (omeprazole and rabeprazole) or the histamine H(2) blocker cimetidine were administered intraperitoneally 1 h before OVA challenge and before measuring cough reflex sensitivity, then bronchoalveolar lavage fluid (BALF) was immediately collected. The pH of the fluid obtained by bronchial washing was determined after examining the effect of rabeprazole on the cough response to capsaicin. RESULTS: The number of coughs elicited by capsaicin was significantly increased 24 h after challenge with OVA compared with saline, indicating antigen-induced increase in cough reflex sensitivity. Both PPIs dose dependently and significantly inhibited antigen-induced cough hypersensitivity. Omeprazole did not influence the antigen-induced increase in the total number of cells or ratio (%) of eosinophils in BALF. Cimetidine did not affect the antigen induced cough hypersensitivity or cellular components of BALF. The pH of the bronchial washing fluid was significantly decreased in antigen-challenged animals. Rabeprazole did not affect the antigen-induced decrease in the pH of bronchial washing fluid. CONCLUSION: These findings show that PPIs, but not histamine H(2) blockers, can directly decrease antigen-induced cough reflex hypersensitivity, while the mechanism remains unclear. PMID- 15784102 TI - The predictive value of specific immunoglobulin E levels in serum for the outcome of oral food challenges. AB - BACKGROUND: Specific serum IgE is considered as one of the important diagnostic measures in the diagnostic work-up of food allergy. OBJECTIVE: To evaluate the role of specific serum IgE in predicting the outcome of oral food challenges, and to determine threshold concentrations of specific serum IgE that could render double-blind, placebo-controlled food challenges unnecessary. METHODS: In 501 children (median age 13 months), 992 controlled oral challenges were performed with cow's milk (CM), hen's egg (HE), wheat and soy. 440/501 (88%) children suffered from atopic dermatitis. For all children, specific IgE concentrations in serum were determined. Sensitivity, specificity, positive and negative predictive values, receiver operator characteristics-curves as well as predictive decision points were calculated. RESULTS: Four hundred and forty-five out of 992 oral food challenges with allergens were assessed as positive. Sensitivity of specific serum IgE was 97% for HE, 83% for CM, 69% for soy, and 79% for wheat. Specificity was 51% for HE, 53% for CM, 50% for soy, and 38% for wheat. Calculating 90%, 95% and 99% predicted probabilities using logistic regression revealed predictive decision points of 6.3, 12.6, and 59.2 kU/L for HE, respectively. Subdividing our children in those of below or above 1 year of age resulted in a markedly different predicted probability for HE. For CM, only the 90% predicted probability (88.8 kU/L) could be calculated. No decision points could be determined for CM, wheat and soy. CONCLUSION: In general, specific serum IgE levels showed a correlation with the outcome of positive oral food challenges for CM and HE. Meaningful predictive decision points can be calculated for HE, which may help to avoid oral food challenges in some cases. However, data need to be ascertained for each allergen separately. Furthermore, the age of the patient population under investigation must also be taken into account. PMID- 15784103 TI - Deficiency in immunoglobulin G2 antibodies against staphylococcal enterotoxin C1 defines a subgroup of patients with atopic dermatitis. AB - BACKGROUND AND AIM: Atopic dermatitis (AD) is a common chronic inflammatory skin disease often accompanied by cutaneous Staphylococcus aureus colonization and, in this regard, especially complicated by the presence of superantigen-producing strains. Because IgG antibodies comprise an important defence mechanism of the adaptive immune system against bacteria, it was investigated whether AD patients have an abnormal pattern or distribution of superantigen-specific IgG subclass antibodies in association with disease severity and activity. METHODS: Staphylococcal enterotoxin B (SEB) and staphylococcal enterotoxin C1 (SEC1) specific IgG antibody subclasses were assessed in n=89 adult AD patients with mild to severe disease activity as determined by the SCORAD score and in n=28 healthy age-matched controls. Results were correlated with the current status of bacterial skin colonization and severity score. RESULTS: Thirty-eight per cent of the AD patients showed a selective deficiency in IgG2 antibodies against SEC1 compared with only 14% in the control group. The absence of these antibodies was found in both currently colonized and non-colonized AD patients and was associated with a severe phenotype (SCORAD more than 40 points in two-thirds of the deficient patients). However, these patients had normal production levels of IgG2 antibodies against pneumococcal capsular polysaccharide (PCP) and SEB, but higher IgG1 and IgG4 titres against SEC1. Except for elevated total IgG1, total IgG subclass levels were normal in this AD subgroup. Yet, peripheral blood mononuclear cells (PBMCs) derived from these patients clearly produced IL-4 and IL-5 upon SEC1 re-stimulation whereas PBMCs from those providing SEC1-specific IgG2 antibodies failed in the production of these cytokines. CONCLUSION: A subgroup of AD patients suffers from a selective deficiency to produce anti-SEC1 IgG2 antibodies. This patient group is characterized by a severe AD phenotype. PMID- 15784104 TI - Effect of a concomitant diagnosis of allergic rhinitis on asthma-related health care use by adults. AB - BACKGROUND: Asthma and allergic rhinitis (AR) frequently coexist, but have usually been studied separately in health economic analyses. OBJECTIVE: To determine the incremental effect of documented AR on health care resource use in adults with asthma. METHODS: A retrospective cohort study using data from a UK general practice database for the period 1998-2001 enrolled subjects 16-55 years of age, with one or more asthma-related general practitioner (GP) visits during a 12-month follow-up period. The study outcomes were asthma-related hospitalizations, GP visits, and prescription drug costs during the 12-month follow-up period for patients with and without physician-diagnosed AR. RESULTS: Concomitant AR was documented in 4,611 (16.9%) of the total sample of 27,303 adults with asthma. Compared with those with asthma alone, patients with concomitant AR experienced more GP visits (5.2 vs. 4.2; P<0.0001) and more of them were hospitalized for asthma (0.76% vs. 0.45%; P<0.01) during the 12-month follow-up period. In multi-variable regression analyses, AR was predictive of hospitalization for asthma (odds ratio 1.52, 95% confidence interval (CI) 1.03 2.24) and was associated with an increase in the annual number of asthma-related GP visits (mean increase per patient 0.42, 95% CI 0.42-0.43) and annual asthma related drug costs (mean increase GBP 5.1, 95% CI 5.0-5.3). CONCLUSION: Adults with asthma and documented concomitant AR experienced more asthma-related hospitalizations and GP visits, and incurred higher asthma drug costs than did adults with asthma alone. A unified treatment strategy for asthma and AR might reduce the costs of treating these conditions. PMID- 15784105 TI - Why generic and disease-specific quality-of-life instruments should be used together for the evaluation of patients with persistent allergic rhinitis. AB - BACKGROUND: The importance of assessing health-related quality of life (HRQL) of patients with allergic rhinitis (AR) has been well established, but the specific roles of rhinitis-specific or general health instruments have not been delineated. OBJECTIVE: We analysed the psychometric properties of a disease specific instrument, the Rhinoconjunctivitis Quality-of-Life Questionnaire (RQLQ) and the general health instrument, the Medical Outcome Short-Form 36 (SF-36) as they are employed in combination in patients with persistent AR in clinical practice. METHOD: We analysed the data collected from a prospective study of 43 newly diagnosed patients with persistent AR and 44 controls. We interviewed the patients four times, at baseline, weeks 4, 8 and 10. RESULTS: The RQLQ and SF-36 have good discriminative property, internal consistency, and test-retest reliability. The RQLQ is superior to the SF-36 as an evaluative instrument because more of its domains respond to change, the magnitude of change was greater, and the response was faster. The SF-36 is more susceptible to floor and ceiling effects. Both instruments are unsuitable for mildly symptomatic patients based on Rasch model analysis. Each questionnaire assesses a distinct and significant portion of the total HRQL of persistent AR. CONCLUSION: The SF-36 and RQLQ are good for discriminating rhinitis patients from controls, but the former is poor for detecting changes in QOL. Both are inappropriate for mildly symptomatic patients. Each instrument measures non-overlapping halves of the measurable HRQL. For an assessment of the HRQL in persistent AR that is complete and responsive both instruments should be employed together. PMID- 15784106 TI - Prevalence of latex allergy in the community at age 7 years. AB - BACKGROUND: Latex allergy has been highlighted as a problem in children during the last decade based on a number of case series of children with particular problems such as spina bifida. The actual prevalence of latex allergy in the general United Kingdom population is unclear. OBJECTIVE: To estimate the prevalence of childhood latex allergy in the general population. METHODS: The Avon Longitudinal Study of Parents and Children is a geographically based cohort that has been prospectively followed since birth. The children were invited for skin prick testing at 7 years of age. RESULTS: Four subjects out of 1877 tested were sensitized to latex. None had a history of clinical reactions to latex. CONCLUSION: This study suggests that the prevalence of latex sensitization and clinical latex allergy in the general childhood population are very low, 0.2% (95% confidence interval 0.1-0.6%) and 0.0% (0-0.2%), respectively. PMID- 15784107 TI - Wheeze, allergic sensitization and geohelminth infection in Butajira, Ethiopia. AB - BACKGROUND: The effect of geohelminth infection on wheeze and allergen sensitization is inconsistent across different epidemiological studies. OBJECTIVE: To investigate the association between self-reported wheeze, self reported asthma, allergic sensitization and geohelminth infection in urban and rural areas of Butajira, southern Ethiopia. METHODS: Questionnaire data on wheeze, asthma and a range of confounding variables was gathered in a cross sectional study of 7649 people aged 5 years or more from the Butajira Rural Health Project database. Allergic skin sensitization to Dermatophagoides pteronyssinus and cockroach was measured, and a stool sample collected for qualitative and quantitative geohelminth analysis. RESULTS: Wheeze was weakly associated with allergic sensitization to D. pteronyssinus and cockroach (odds ratios (OR) 1.21, 95% confidence interval (CI) 0.98-1.51, and 1.27, 95% CI 1.00 1.62, respectively). Self-reported asthma was related to sensitization to D. pteronyssinus only (OR 4.09, 95% CI 2.86-5.84). Geohelminths were present in 33.8% of participants, and the median egg load in infested individuals was 6 eggs/g. Overall, presence of any geohelminths was associated with a diminished risk of cockroach sensitization (adjusted OR 0.82, 95% CI 0.68-0.99) but there were no significant protective effects of any geohelminth infection against wheeze or asthma. CONCLUSION: In a developing country community with relatively low geohelminth prevalence and intensity, we found weak association between allergic sensitization and wheeze, but no evidence of a protective effect of geohelminths against wheeze or asthma. PMID- 15784108 TI - Increase in daytime symptoms is a sensitive and specific criterion for predicting corticosteroid-treated exacerbations in a clinical asthma trial. AB - BACKGROUND: To determine which diary card variables are the most predictive for administration of additional courses of corticosteroids using the TRUST (The Regular Use of Salbutamol Trial) data set. METHODS: Logistic regression models were used to identify the extent to which a change in diary card variable affected the odds ratio (OR) for administering a course of oral or increased inhaled corticosteroids. The complete TRUST diary card data were used with over 200,000 days of diary card observations from 983 mild to moderate asthmatic subjects. RESULTS: An increase in daytime symptoms of 1-5 U over baseline was associated with an increase in the OR for starting all types of corticosteroids from two- to 60-fold. CONCLUSIONS: These results indicate that an increase in daytime symptoms of two or more over baseline strongly predicts the administration of additional corticosteroids. The results have significant implications for both clinical practice and design of clinical trials in asthma. PMID- 15784109 TI - Bilateral nasal allergen provocation monitored with acoustic rhinometry. Assessment of both nasal passages and the side reacting with greater congestion: relation to the nasal cycle. AB - BACKGROUND: The effect of bilateral nasal provocation on nasal mucosa measured with the use of acoustic rhinometry (AR) can be assessed for both nasal passages or for the side responding with greater congestion. Assessment of changes in nasal congestion during the nasal provocation test (NPT) can be affected by the nasal cycle (NC). The aim of this study was to find out the most accurate method to evaluate changes observed during bilateral nasal provocation. METHODS: Cross sectional areas (CSA) at the level of inferior nasal turbinate (CSA-2) were recorded by AR in 26 volunteers with allergic rhinitis during the NC for 5-7 h and subsequently during NPT. The risk of spontaneous total and unilateral CSA-2 decrease was established. Sensitivity of the NPT assessment for the total CSA-2 and for the side responding with greater congestion was evaluated at chosen thresholds. These thresholds were selected in a way that the risk levels of spontaneous decrease of unilateral and total CSA-2 were equal. RESULTS: The assessment of the total CSA-2 was found to be more sensitive than the assessment of the side responding with greater congestion. The highest sensitivity and specificity of the test was achieved by using a combination of both assessments. Optimum thresholds of the CSA-2 decrease for assessment at 15 min after provocation, with this method, were 27% and 40% for the side responding with greater congestion and for the total CSA-2, respectively. CONCLUSIONS: Recognition of the risk of spontaneous unilateral and total CSA-2 decreases enables introduction of combined assessment of bilateral NPT. This assessment seems to be the most accurate method for evaluation of the test results. PMID- 15784110 TI - Airway inflammation in nasal polyposis: immunopathological aspects of relation to asthma. AB - BACKGROUND: Nasal polyposis (NP) is a chronic inflammatory disorder of the upper respiratory tract, which is often coexist with asthma. However, the pathogenesis of especially in patients with NP is still a matter of debate. OBJECTIVE: To better understand the immunopathologic mechanism involved in this relationship, we investigated the inflammatory cell profiles in bronchial and nasal tissues of patients with NP alone and with concomitant asthma. METHODS: Seventeen patients with NP (six male, 11 female, age range: 19-63, mean age: 38.29+/-13.27 years) were selected for the study. Subjects were divided into two groups based on the presence of asthma or bronchial hyper-responsiveness (BHR). NP without BHR (Group 1) (n=8), NP and asthma or BHR (Group 2) (n=9). All patients underwent atopy evaluation including detailed history, skin prick test (SPT), total and specific IgE determination in sera. None of the subjects had taken inhaled, nasal or oral corticosteroids for at least 1 month before the study. Respiratory symptoms of asthmatic patients were controlled with only short acting beta(2)-agonist inhaler drugs as needed. NP tissue, nasal and bronchial mucosa biopsies were taken from all patients using fiberoptic endoscopy. CD3, CD8, CD16, CD68, AA1 (mast cell tryptase), human leucocyte antigen-DR (HLA-DR) and eosinophil peroxidase (EPO) expressing cells in specimens were determined by immunohistochemical methods. Positively staining inflammatory cell types were counted. Subepithelial lamina propria and periglandular areas were separately evaluated. RESULTS: No significant difference was found in polyp tissue, nasal and bronchial CD3(+), CD8(+), CD16(+), CD68(+), AA1(+), HLA-DR(+) and EPO(+) positive cells between groups. There were significantly higher numbers of CD8(+), CD16(+), HLA-DR(+), EPO(+) cells in the polyp tissue and nasal mucosa vs. the bronchial mucosa in all groups (P<0.05). However, CD8(+) cells were significantly increased in the polyp tissue and bronchial mucosa of patients with NP alone when compared with the patients with both asthma and NP (P<0.05). CD3(+), CD68(+) and CD16(+) cell counts were tended to be higher within the nasal polyp tissue of patients with isolated NP compared with counts within nasal and bronchial mucosa of patients with NP and asthma. Also, patients with isolated NP showed more HLA-DR(+) cells in the nasal polyp tissue and nasal mucosa than those of patients with NP and asthma. Immunoreactivity for EPO(+) eosinophils within the nasal and bronchial mucosa was more prominent in patients with NP and asthma compared with patients with NP alone. The number of EPO(+) eosinophils within the polyp tissue, nasal and bronchial mucosa was higher in the skin prick test negative (SPT -ve) group than the SPT positive (SPT +ve) ones. CONCLUSIONS: Our results demonstrate that infiltration of inflammatory cells in the nasal and the lower airways do not remarkably differ between patients with NP alone who has no evidence of BHR and asthmatic patients with NP. However, patients with SPT-ve NP reveal more intense eosinophilic inflammation in the entire respiratory mucosa. PMID- 15784111 TI - A polymorphism in the promoter region of the human interleukin-16 gene is not associated with asthma or atopy in an Australian population. AB - BACKGROUND: IL-16 is an immunomodulatory cytokine whose expression is increased in the bronchial mucosa, bronchoalveolar lavage fluid and induced sputum of asthmatic patients. It has been suggested that IL-16 has a regulatory role in the pathophysiology of asthma. A single-nucleotide polymorphism (T(-295)C) has been described in the promoter region of the gene and it has been hypothesized that this polymorphism may be associated with altered levels of IL-16 expression, and account for the increased levels of IL-16 seen in the asthmatic airway. OBJECTIVE: To determine the association between the T(-295)C promoter polymorphism and asthma, disease severity and atopy in a large Australian Caucasian population. METHODS: We used PCR and restriction fragment length polymorphism analysis to establish the allele frequency of the T(-295)C promoter polymorphism in a random Australian Caucasian population (n=176) and to characterize the polymorphism in a large Australian Caucasian population of mild (n=273), moderate (n=230) and severe (n=77) asthmatic patients, and non-asthmatic controls (n=455). Genotype association analyses were performed using chi(2) tests. RESULTS: The polymorphic C allele was present in 19% of the asthmatic population and 21% of the non-asthmatic population. There was no association between the polymorphism and asthma (P=0.153) nor with asthma severity (P=0.417) or atopy (P=0.157) in this population. CONCLUSION: Although it has been hypothesized that the T(-295)C promoter polymorphism may be associated with increased IL-16 gene expression, it is not associated with asthma, disease severity or atopy in this Australian population. PMID- 15784112 TI - Polymorphisms in the 5-lipoxygenase activating protein (ALOX5AP) gene are not associated with asthma in an Australian population. AB - BACKGROUND: The cysteinyl-leukotrienes (cys-LTs) are important pro-inflammatory mediators in asthma, and have been shown to have a role in specific disease subtypes, including asthma severity. Few studies have investigated the role of polymorphisms in the ALOX5AP gene, encoding 5-lipoxygenase activating protein (FLAP), and asthma. We hypothesized that polymorphisms in this gene are associated with asthma and in particular, with asthma severity, in an Australian population. OBJECTIVE: To screen the coding region of the ALOX5AP gene for polymorphisms and to determine the association between previously described polymorphisms and asthma and asthma severity in an Australian population. METHODS: We used PCR-SSCP and PCR-RFLP analysis to examine a previously described promoter polyA variable repeat polymorphism and two intronic polymorphisms (IVS2+12C>A, IVS2+105T>C), and to screen all five exons of the gene for new polymorphisms, in a large Australian population of randomly selected, non asthmatic controls (n=457), mild asthmatics (n=274), moderate asthmatics (n=231) and severe asthmatics (n=79). RESULTS: We confirmed the presence of two polymorphisms in intron 2 and found no association between these polymorphisms and asthma or asthma severity, nor between a promoter polymorphism in the ALOX5AP gene and asthma or asthma severity. Gene fragment analysis of the promoter polymorphism revealed novel, conserved repeat numbers in our population, and no new polymorphisms were found in the coding region of the gene. CONCLUSION: These findings in a large, well characterized asthma population, reveal that, while FLAP is an important enzyme in cys-LTs biosynthesis, polymorphisms in the ALOX5AP gene are not likely to be functionally associated with the asthma phenotype. PMID- 15784113 TI - The human leucocyte antigen-DRB1*1302-DQB1*0609-DPB1*0201 haplotype may be a strong genetic marker for aspirin-induced urticaria. AB - BACKGROUND: Urticaria/angioedema is a common aspirin-induced allergy; however, its pathogenic mechanism is not understood. OBJECTIVE: In order to uncover the genetic mechanism, we studied the associations of the human leucocyte antigen (HLA) genotypes in patients with aspirin-induced urticaria compared with aspirin intolerant asthma and normal control in a Korean population. METHODS: Ninety-four aspirin-induced urticaria patients presenting urticaria/angioedema-induced by both ASA and NSAID (50 had underlying chronic urticaria) and showing positive responses on oral aspirin challenge test, 76 aspirin-intolerant asthmatics with positive responses on lysine-aspirin bronchoprovocation test, and 185 normal healthy controls were enrolled. HLA-DRB1, DQB1, and DPB1 genotypings were performed by direct DNA sequencing analysis. RESULTS: The allele frequencies of HLA-DRB1(*)1302 (18.1%) and HLA-DQB1(*)0609 (10.1%) in aspirin-induced urticaria were significantly higher than in aspirin-intolerant asthma (5.3%, P=0.0004; 2.0%, P=0.0024) and in normal controls (8.1%, P=0.0005; 3.2%, P=0.0008), and they remained significant after correcting for multiple comparisons. The patients with these two HLA markers had a significantly younger age than patients without, while no associations were found in with respect to atopic status, a history of previous allergic diseases, total IgE level, or presence of underlying chronic urticaria (P>0.05, respectively). In haplotype analysis, the HLA-DRB1(*)1302 DQB1(*)0609-DPB1(*)0201 was significantly higher in the aspirin-induced urticaria (8.0%) than in the aspirin-intolerant asthma (0.7%, P=0.0014) and normal controls (2.0%, P=0.0006). CONCLUSION: These findings suggest that the HLA-DRB1(*)1302 DQB1(*)0609-DPB1(*)0201 may be a strong genetic marker to determine the aspirin induced urticaria phenotype. PMID- 15784114 TI - 1,3-beta-glucanases as candidates in latex-pollen-vegetable food cross reactivity. AB - BACKGROUND: 1,3-beta-glucanases (group 2 of pathogenesis-related proteins) are enzymes widely distributed among higher plants and have been recently proven to be significant allergens. OBJECTIVE: The aim of this work was to study the potential implication of 1,3-beta-glucanases in cross-reactivities among latex, pollen and vegetable foods. METHODS: The cDNA encoding the N-terminal domain (NtD) of Ole e 9, a major allergenic 1,3-beta-glucanase from olive pollen, was amplified by polymerase chain reaction and produced as a recombinant protein in Pichia pastoris (recombinant N-terminal domain, rNtD). Circular dichroism, ELISA, immunoblotting and immunoblotting inhibition experiments were carried out. Sera from olive pollen allergic patients and a rNtD-specific polyclonal antiserum were used. RESULTS: The NtD of Ole e 9 has been produced at high yield in the yeast P. pastoris and possesses 1,3-beta-glucanase activity. The expressed polypeptide conserves IgE and IgG immunodominant epitopes of the whole Ole e 9. A rNtD specific polyclonal antiserum and sera from olive pollen allergic patients allowed detection of IgG and IgE reactive peptidic epitopes common to 1,3-beta glucanase Ole e 9 in extracts from ash and birch pollen, tomato, potato, bell pepper, banana and latex. CONCLUSION: rNtD and homologous glucanases are new molecules to be used in diagnostic protocols as they could help to identify allergic pollen patients who are at risk for developing allergic symptoms to fruits, vegetables and latex. PMID- 15784115 TI - Glutathione protects human airway proteins and epithelial cells from isocyanates. AB - BACKGROUND: Glutathione (GSH), one of the major anti-oxidants of the lung, has been linked to the human response to isocyanate exposure. However, the ability of GSH to modulate key chemical reactions, thought to be central to the development of human isocyanate allergy, has not been directly analyzed under biologic exposure conditions. OBJECTIVE: To better understand the potential role of GSH in the response to occupational isocyanate exposure, we evaluated its effects on two processes thought to be involved in the development of isocyanate allergy, isocyanate-protein conjugation and epithelial cell toxicity. METHODS: The effects of GSH on (1) isocyanate conjugation with albumin, its major target in the airway fluid and (2) isocyanate-induced toxicity to human airway epithelial cell lines, A549 and NCI-H292, were tested using two different in vitro models. For protein conjugation studies, a newly described vapour exposure system was used to model the air/liquid interface at the surface of the epithelial fluid in the airways. Epithelial cell exposures were performed in fluid phase to mimic the in vivo exposure of airway cells covered by epithelial lining fluid. RESULTS: Reduced GSH prevented hexamethylene diisocyanate (HDI) conjugation to albumin in a dose dependent manner, while oxidized GSH (GSSG) conversely increased conjugation rates. GSH levels equivalent to those found in normal human airway fluid (100 microm) provided >90% protection against HDI-protein conjugation when albumin was exposed to HDI vapour levels 10-fold above permissible occupational limits. Physiologic levels of GSH, but not GSSG, also reduced HDI toxicity to human airway epithelial cells in a dose-dependent manner, when present extracellularly, however, drugs that modulate intra-cellular GSH levels did not significantly alter isocyanate toxicity. CONCLUSIONS: Together with previously reported genetic and toxicity studies, the data suggest that airway GSH plays an important role in protection against HDI exposure and may help prevent the development of allergic sensitization and asthma. PMID- 15784116 TI - Cytosine-phosphate-guanine motifs fail to promote T-helper type 1-polarized responses in human neonatal mononuclear cells. AB - BACKGROUND: The T-helper type 1 (Th1) trophic properties of bacterial cytosine phosphate-guanine (CpG) motifs have made them logical adjuvants both for the suppression of Th2-mediated allergic disease in early life and for promoting vaccine responses in neonates who have relatively immature Th1 function. However, little is known about their effects on immature immune responses in this period. OBJECTIVES: To compare the effects of CpG on adult and neonatal cellular immune responses to various stimuli. METHODS: The immune responses of mononuclear cells (MC) derived from neonates (n=25) and their mothers (n=25) were compared in vitro. These were stimulated with house dust mite (HDM), CpG B, CpG C, non-CpG oligodeoxynucleotides (ODN) or diphtheria toxoid (DT) in optimized conditions. In parallel cultures, CpGs were combined with HDM or DT antigens to assess the effect of the various ODN on these antigen-specific responses. Lymphoproliferation and cytokine responses IL-13, IFN-gamma, IL-6, IL-10, TNF alpha) were measured for all of the cultures described above. RESULTS: Although neonates showed attenuated lymphoproliferation to CpG, the production of antigen presenting cell-derived cytokines such as IL-6 and IL-10 and the up-regulation of major histocompatibility complex class II (HLA-DR) were detected at adult levels. T cell-derived cytokines (IL-13 and IFN-gamma) were not detectable in response to CpG alone. Most neonates also failed to produce detectable IFN-gamma to HDM or DT (unlike adults), but readily produced IL-13 to these stimuli. The addition of CpG resulted in an increase in neonatal IFN-gamma production in response to HDM (P=0.011) and a similar but non-significant trend with DT. However, rather than inhibiting Th2 IL-13 responses, the addition of CpGs was associated with a significant increase in the IL-13 responses to HDM (P=0.016) and DT (P=0.03), effects not seen in adults. CONCLUSIONS: This study provides further evidence that neonatal MC responses to CpG are functionally different from adults, and do not show clear Th1 polarization. The CpG associated increase in Th2 responses may reflect a potentiation of the normal neonatal Th2 propensity, or non-specific activation of neonatal MC. PMID- 15784117 TI - Characterization of the human T cell response to antigen 5 from Vespula vulgaris (Ves v 5). AB - BACKGROUND: The T cell reactivity to the major allergen of bee venom, phospholipase A2, has been thoroughly characterized. In contrast, only little is known about the human cellular response to major allergens from wasp venom. OBJECTIVE: To characterize the human T cell response to antigen 5 from Vespula vulgaris, Ves v 5. METHODS: Recombinant Ves v 5 was used to establish allergen specific T cell lines (TCL) and T cell clones (TCC) from the peripheral blood of vespid-allergic and non-allergic individuals. Ves v 5-specific TCL were mapped for T cell epitopes using overlapping synthetic peptides representing the complete amino acid sequence of Ves v 5. Ves v 5-specific TCC were analysed for antigen-induced secretion of IL-4, IFN-gamma and IL-10. RESULTS: Seventeen distinct T cell epitopes were recognized by allergic individuals among which Ves v 5(181-192) was identified as a dominant T cell epitope. Partially different epitopes were observed in TCL from non-allergic subjects and the dominant epitope Ves v 5(181-192) was not prevalent in these cultures. Ves v 5-specific TCC isolated from allergic individuals did not show the typical T helper type 2 (Th2) like cytokine profile in response to specific stimulation, i.e. high amounts of IL-4 and low IFN-gamma. TCC from non-allergic individuals showed a Th1-like cytokine pattern. CONCLUSIONS: Our findings provide evidence that the allergic T cell response to Ves v 5 is not Th2-dominated and that different immunogenic sites on this major wasp venom allergen are recognized by allergic and non allergic individuals. PMID- 15784118 TI - Pho d 2, a major allergen from date palm pollen, is a profilin: cloning, sequencing, and immunoglobulin E cross-reactivity with other profilins. AB - BACKGROUND: Up to now, some date palm pollen (DPP) allergens have been described but very few data are available about their molecular nature. The aim of this study was to identify and characterize Pho d 2, a major allergen from this pollen. METHODS: Sera from 25 patients allergic to DPP were analysed by immunoblotting. Purification of DPP profilin was performed by poly-l-proline affinity chromatography. Profilin-encoding cDNA from DPP was cloned by using a RT PCR strategy and recombinant allergen was expressed as a non-fusion protein in Escherichia coli. Natural and recombinant Pho d 2 were investigated by means of enzyme allergosorbent test to compare the immunologic properties of both allergens and to analyse cross-reactivity with other profilins. RESULTS: A 14.4 kDa protein was identified as a major allergen in DPP extract. Purification, cloning, heterologous expression, and inhibition experiments identified it as profilin (Pho d 2). Pho d 2 comprises 131 amino acids and has high sequence identity with other allergenic food and pollen profilins. The prevalence of specific IgE antibody reactivity to natural Pho d 2 by ELISA was 56% and 64% by skin prick test (SPT). Pho d 2 is an important allergen as it is responsible for more than 70% of the IgE reactivity to the pollen extract. IgE directed against Pho d 2 showed a strong cross-reactivity with other profilins such as those from olive tree and grass pollens. CONCLUSION: Pho d 2, a 14.4 kDa protein identified as profilin, is a major and relevant allergen in DPP, as confirmed by SPT and thereby may elicit clinical symptoms in sensitized patients. PMID- 15784119 TI - Percutaneous application of peptidoglycan from Staphylococcus aureus induces an increase in mast cell numbers in the dermis of mice. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease with immunopathologic features that vary depending on the duration of the lesion. The dermis of lesional skin of AD patients shows an increased number of inflammatory cells such as mast cells, eosinophils and mononuclear cells and superficial Staphylococcus aureus colonization. OBJECTIVE: The purpose of this study was to determine the effects of peptidoglycan (PEG) from S. aureus on mast cell induction in murine skin. METHODS: PEG was applied to barrier-disrupted abdominal skin of mice every 5 days and the number of mast cells in the abdominal skin was counted 20 days after the first application. The cytokine response was investigated by RT-PCR and immunohistologic analysis. RESULTS: The number of mast cells in the skin of mice treated with PEG was increased significantly compared with that of mice given phosphate-buffered saline. In addition, application of PEG to the abdominal skin increased the expression of mRNA for transforming growth factor-beta(1) (TGF-beta(1)), which supports mast cell migration, but not that for IL-3 or stem cell factor, which support both mast cell proliferation and mast cell migration. Immunohistologic analysis demonstrated that levels of TGF beta(1) transcripts corresponded with those of protein synthesis in the epidermis. TGF-beta(1) was found to be highly expressed in keratinocytes of the basal epidermis of PEG-treated skin. Furthermore, intraperitoneal injection of anti-TGF-beta(1) antibodies neutralized the induction of mast cells into the skin. CONCLUSION: These results suggest that PEG may have the ability to induce an increase in mast cell numbers in the skin through TGF-beta(1) production by epidermal keratinocytes. Skin inflammation might therefore be linked to colonization with S. aureus in AD patients. PMID- 15784121 TI - Prenatal lipopolysaccharide-exposure prevents allergic sensitization and airway inflammation, but not airway responsiveness in a murine model of experimental asthma. AB - BACKGROUND: Epidemiological evidence underlines the impact of prenatal environmental factors on the development of postnatal allergies. In this regard an inverse correlation between lipopolysaccharide (LPS) exposure and development of childhood allergy has been found. OBJECTIVE: To assess the impact of prenatal LPS exposure on the development of postnatal respiratory allergies in a mouse model of experimental asthma. METHODS: Female BALB/c mice were exposed to LPS before conception and during pregnancy. Several weeks after birth offspring were sensitized to ovalbumin (OVA) followed by aerosol allergen challenges. RESULTS: Prenatal, maternal LPS-exposure enhanced neonatal IFN-gamma, but not IL-4 and IL 2 production. OVA sensitization of prenatally LPS-exposed mice was accompanied by a marked suppression in anti-OVA IgG1 and IgE as well as unchanged IgG2a antibody responses, paralleled by a significant reduction in IL-5 and IL-13 levels following mitogenic stimulation of splenic leucocytes. Assessment of bronchoalveolar lavage fluids following allergen challenges revealed a marked reduction in eosinophils and macrophages in these mice. Surprisingly, development of airway hyper-responsiveness, a hallmark of bronchial asthma, was not affected. CONCLUSION: This study provides first experimental evidence that LPS may already operate in prenatal life in order to modulate the development of allergies in the offspring. PMID- 15784122 TI - Thrombelastography/thromboelastometry. AB - The term thrombelastograph (TEG) was used to describe the trace produced from the measurement of the viscoelastic changes associated with fibrin polymerization. Recently the term rotational thromboelastometry has been applied to the output of the ROTEM instrument. Since its first description in 1948, the TEG/ROTEM has been successfully used in the near patient assessment of haemostasis. The greatest use has been the application of TEG-guided transfusion of blood components in hepatic and more widely in cardiac surgery. Recent years have seen a renewed interest in the technology with applications for both pharmaceutical monitoring and patient screening being described. The present review gives a broad overview of the developments and applications related to thrombelastography/thromboelastometry. PMID- 15784120 TI - Therapeutic administration of Budesonide ameliorates allergen-induced airway remodelling. AB - BACKGROUND: Airway inflammation and remodelling are important pathophysiologic features of chronic asthma. Although current steroid use demonstrates anti inflammatory activity, there are limited effects on the structural changes in the lung tissue. OBJECTIVE: We have used a mouse model of prolonged allergen challenge that exhibits many of the salient features of airway remodelling in order to investigate the anti-remodelling effects of Budesonide. METHODS: Treatment was administered therapeutically, with dosing starting after the onset of established eosinophilic airway inflammation and hyper-reactivity. RESULTS: Budesonide administration reduced airway hyper-reactivity and leukocyte infiltration in association with a decrease in production of the Th2 mediators, IL-4, IL-13 and eotaxin-1. A reduction in peribronchiolar collagen deposition and mucus production was observed. Moreover, our data show for the first time that, Budesonide treatment regulated active transforming growth factor (TGF)-beta signalling with a reduction in the expression of pSmad 2 and the concomitant up regulation of Smad 7 in lung tissue sections. CONCLUSIONS: Therefore, we have determined that administration of Budesonide modulates the progression of airway remodelling following prolonged allergen challenge via regulation of inflammation and active TGF-beta signalling. PMID- 15784123 TI - Evaluation of the monocyte counting by two automated haematology analysers compared with flow cytometry. AB - The aim is to determine the monocyte count performance of the Bayer Diagnostics ADVIA120 and Coulter LH 750 automated haematology analysers and the results obtained by these two instruments were compared with those provided by Becton Dickinson FACScan flow cytometer using the combination of CD45/CD14 MoAb. Linearity and imprecision were also evaluated. The linearity of both instruments was good. Coulter LH 750 showed better precision (4.3%) than ADVIA 120 (9.0%) both within and between batch. A significant correlation (r = 0.973) was found between the LH 750 and the flow cytometry method, while a modest one was observed between the latter and the ADVIA 120 (r = 0.880). When comparing the percentage of monocytes by means of one-way anova and Tukey test, it was found that the LH 750 provided the closest results in comparison with flow cytometry, with no statistical difference between the means (mean difference MO% = 0.6); however the difference was statistically different between the ADVIA 120 and flow cytometry (mean difference MO% = -4.06). These data were confirmed by Altman-Bland and Deming regression analyses. PMID- 15784124 TI - Utility of point-of-care haemoglobin measurement in the HemoCue-B haemoglobin for the initial diagnosis of anaemia. AB - Measurement of haemoglobin (Hb) concentration provides a reliable indication of the presence and severity of anaemia. However, other laboratory parameters are usually requested as well, leading to an increase in socio-sanitary costs. Accordingly, this study was undertaken to ascertain the reliability of point-of care Hb determination with the portable photometer HemoCue-B haemoglobin (HBH) and to evaluate its utility for the initial diagnosis of anaemia. Hb was measured (x3) in 20 venous blood samples diluted with saline (v/v; 1 : 0, 2 : 1, 1 : 1, 2 : 1 and 3 : 1) to obtain a wide range of Hb and in venous and capillary blood samples from 247 primary health care patients. All HBH results were compared with those yielded by the reference cell counter Pentra 120 Retic (ABX). In diluted samples, Hb values obtained with either method were not significantly different (ABX-HBH, -0.01 +/- 0.32 g/dl; 95% CI, -0.04 to 0.028 g/dl) and showed an excellent Pearson's coefficient of correlation (r = 0.992; P < 0.01). HBH provides accurate values if at least 4 mul of blood is loaded into the cuvette. There were no significant differences between Hb measured in venous (v) and capillary (c) blood samples in primary care patients. Eighteen anaemic patients were detected by ABX measurements (7.3%; 15 female/3 male), 18 by HBHv (specificity, 100%; sensitivity, 100%) and 25 by HBHc (eight false positives; one false negative; specificity, 94.4%; sensitivity, 96.5%). Compared with ABX, HBH provides accurate and precise measurements for a wide range of Hb and its use in primary health care seems to be a good method for the initial diagnosis of anaemia. PMID- 15784125 TI - Blood lead levels in iron-deficient and noniron-deficient adults. AB - Iron deficiency (ID) has been reported to increase lead absorption. This relationship has been investigated in detail in children but not in adults. This study was designed to investigate whether blood lead levels are significantly higher in iron-deficient adults. ID-parameters (haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin, serum iron, total iron-binding capacity, iron-binding saturation, soluble transferrin receptors, washed zinc protoporphyrin and ferritin) together with whole blood lead were measured in three different adult groups - blood donors (n = 73), pregnant women (n =74) and haemodialysis patients (n = 72). Of a total of 219 subjects tested, 7.7% was found to have a lead level above 10 microg/dl (maximum 16 microg/dl). No association was found between blood lead level and ID [iron-deficient subjects (n: 139), mean: 5.6 microg/dl (SD: 3 microg/dl) and noniron-deficient subjects (n: 80), mean: 5.4 microg/dl (SD: 3 microg/dl)]. The results suggest that the inverse association between blood lead and serum iron in studies carried out on children does not occur in adults. PMID- 15784126 TI - High pressure liquid chromatography and electrospray ionization mass spectrometry are advantageously integrated into a two-levels approach to detection and identification of haemoglobin variants. AB - Detecting and correctly identifying haemoglobin (Hb) variants is typically achieved by a two-levels laboratory approach. We report our experience in dealing with 91 Hb variants, including a number of frequent and a few rare variants. Screening included akaline agarose gel electrophoresis (AGE), ion-exchange automated high-performance liquid chromatography (HPLC) and a test for deoxyhaemoglobin solubility. Identification was based on electrospray ionization mass spectrometry (ESI-MS). Our results confirmed the advantages of HPLC over AGE for screening, because of the occurrence of some electrophoretically 'silent' variants. ESI-MS permitted the definitive identification of 90 of the 91 variants included in the study, in some cases (e.g. HbS) through the application of a simple protocol (direct injection of the sample), in other cases requiring the application of more demanding procedures (purification of the variant chain and peptide analysis after enzymatic or chemical cleavage). In an additional case (Hb J-Oxford), ESI-MS assay did not lead to definitive identification, but gave indications for designing the appropriate primers to focus DNA sequence analysis on the specific region of the gene. Deoxyhaemoglobin solubility test was positive only in the presence of HbS. We conclude that HPLC and ESI-MS are advantageously integrated into a two-level analytical system for the detection and confirmation of variant Hbs. PMID- 15784127 TI - Thrombotic thrombocytopenic purpura in southern Turkey: a single-center experience of 29 cases. AB - In this retrospective, nonrandomized study, we describe our experience in the management of 29 consecutive patients with thrombotic thrombocytopenic purpura (TTP) treated with a combined therapy of plasma exchange (PE) and steroids at a single center. We compared the effectiveness of high-dose steroids (20-25 mg/kg methyl prednisolone) as first-line treatment in combination with PE therapy with the combination of standard-dose steroids (1 mg/kg methyl prednisolone) and PE in adult patients with TTP. Clinical, laboratory data and treatment outcomes such as response rate, median time to recovery and survival were evaluated retrospectively. Overall (OR) and complete (CR) response rates were 69 and 52% respectively. Similar response rates were found in patients treated with pulse or conventional dose steroids; however, the median time to response was delayed in the high-dose methyl prednisolone (HDP) group. The median time from the initiation of symptoms to initiation of treatment was approximately 15 days (range: 0-30). Delayed treatment in our patients because of delayed referral to our center resulted in poor response to treatment. In all, four of 14 (27%) complete responders experienced relapses. The predicted relapse rate was 48% at a median of 30 months. All the relapses presented with a combination of thrombocytopenia and microangiopathic hemolytic anemia. This analysis showed that high-dose steroid treatment did not prove to be beneficial for TTP patients as firstline therapy combined with PE. Moreover, pulse steroid interventions might have resulted in delayed responses and our data suggest that initiation of treatment with PE should not be delayed. PMID- 15784128 TI - The role of RhD agglutination for the detection of weak D red cells by anti-D flow cytometry. AB - Anti-D flow cytometry is an accurate method for quantifying feto-maternal haemorrhage (FMH). However, weak D red cells with <1000 RhD sites are not detectable using this methodology but are immunogenic. As quantitation of RhD sites is not practical, an alternative approach is required to identify those weak D fetal red cells where anti-D flow cytometry is inappropriate. We describe a simple algorithm based on RhD agglutination and flow cytometry peak separation. All weak D (n = 34) gave weak agglutination with RUM-1 on immediate spin (grading 95% proteins expressed in trans-differentiated cells could be associated with the development, differentiation and regulation of nervous system cells. CONCLUSION: Our results indicate that the cat melanoma cells have the ability to differentiate into distinct neuronal cell types and they express proteins that are essential for self-renewal. Since melanocytes arise from the neural crest of the embryo, we conclude that this melanoma arose from embryonic precursor stem cells. This model system provides a unique opportunity to identify domains of interactions between the expressed proteins that halt the tumorigenic potential of melanoma cells and drive them toward neurogenerative pathways involved in early neurogenesis. A better understanding of these proteins in a well coordinated signaling network would also help in developing novel approaches for suppression of highly malignant tumors that arise from stem-like embryonic cells. PMID- 15784143 TI - Origin-independent plasmid replication occurs in vaccinia virus cytoplasmic factories and requires all five known poxvirus replication factors. AB - BACKGROUND: Replication of the vaccinia virus genome occurs in cytoplasmic factory areas and is dependent on the virus-encoded DNA polymerase and at least four additional viral proteins. DNA synthesis appears to start near the ends of the genome, but specific origin sequences have not been defined. Surprisingly, transfected circular DNA lacking specific viral sequences is also replicated in poxvirus-infected cells. Origin-independent plasmid replication depends on the viral DNA polymerase, but neither the number of additional viral proteins nor the site of replication has been determined. RESULTS: Using a novel real-time polymerase chain reaction assay, we detected a >400-fold increase in newly replicated plasmid in cells infected with vaccinia virus. Studies with conditional lethal mutants of vaccinia virus indicated that each of the five proteins known to be required for viral genome replication was also required for plasmid replication. The intracellular site of replication was determined using a plasmid containing 256 repeats of the Escherichia coli lac operator and staining with an E. coli lac repressor-maltose binding fusion protein followed by an antibody to the maltose binding protein. The lac operator plasmid was localized in cytoplasmic viral factories delineated by DNA staining and binding of antibody to the viral uracil DNA glycosylase, an essential replication protein. In addition, replication of the lac operator plasmid was visualized continuously in living cells infected with a recombinant vaccinia virus that expresses the lac repressor fused to enhanced green fluorescent protein. Discrete cytoplasmic fluorescence was detected in cytoplasmic juxtanuclear sites at 6 h after infection and the area and intensity of fluorescence increased over the next several hours. CONCLUSION: Replication of a circular plasmid lacking specific poxvirus DNA sequences mimics viral genome replication by occurring in cytoplasmic viral factories and requiring all five known viral replication proteins. Therefore, small plasmids may be used as surrogates for the large poxvirus genome to study trans-acting factors and mechanism of viral DNA replication. PMID- 15784144 TI - Modulation of macrophage functions by sheeppox virus provides clues to understand interaction of the virus with host immune system. AB - BACKGROUND: Poxviruses encode a range of immunomodulatory genes to subvert or evade the challenges posed by the innate and adaptive immune responses. However, the inactivated poxviruses possessed immunostimulating capacity and were used as a prophylactic or metaphylactic application that efficiently reduced susceptibility to infectious diseases in different species. This fact is intensively studied in different genera of poxviruses. However, little is known about the basic mechanisms adopted by sheeppox virus (SPPV). SPPV causes an acute disease of sheep that recently, has been observed to reinfect its host in spite of vaccination. RESULTS: By injecting inactivated or attenuated sheeppox virus SPPV vaccine in adult male Swiss mice, SPPV was found to reduce macrophages' functions in a local event that occurs at the site of application 12 h after vaccine administration as indicated by increased level of IL-10 and decreased level of SOD from cultured peritoneal macrophages. In contrast increased levels of IL-12, and SOD activity from cultured splenic macrophages, lymphocyte response to PHA-P, and in-vivo response to T-dependant Ag were detected. These effects were observed in both attenuated and inactivated SPPV, but more prominent in attenuated one. CONCLUSION: The results of this study help to elucidate, the phenomenon of existence natural SPPV infections in sheep instead of vaccination and the basic mechanisms responsible for the immunostimulating capacity of sheeppox virus. Locally, SPPV shows evidence for an immune escape mechanism that alleviates the host's immune response. Later and systemically, the virus protects the host from any fatal consequences of the immune system suppression. PMID- 15784145 TI - Molecular biodiversity of cassava begomoviruses in Tanzania: evolution of cassava geminiviruses in Africa and evidence for East Africa being a center of diversity of cassava geminiviruses. AB - Cassava is infected by numerous geminiviruses in Africa and India that cause devastating losses to poor farmers. We here describe the molecular diversity of seven representative cassava mosaic geminiviruses (CMGs) infecting cassava from multiple locations in Tanzania. We report for the first time the presence of two isolates in East Africa: (EACMCV-[TZ1] and EACMCV-[TZ7]) of the species East African cassava mosaic Cameroon virus, originally described in West Africa. The complete nucleotide sequence of EACMCV-[TZ1] DNA-A and DNA-B components shared a high overall sequence identity to EACMCV-[CM] components (92% and 84%). The EACMCV-[TZ1] and -[TZ7] genomic components have recombinations in the same genome regions reported in EACMCV-[CM], but they also have additional recombinations in both components. Evidence from sequence analysis suggests that the two strains have the same ancient origin and are not recent introductions. EACMCV-[TZ1] occurred widely in the southern part of the country. Four other CMG isolates were identified: two were close to the EACMV-Kenya strain (named EACMV-[KE/TZT] and EACMV-[KE/TZM] with 96% sequence identity); one isolate, TZ10, had 98% homology to EACMV-UG2Svr and was named EACMV-UG2 [TZ10]; and finally one isolate was 95% identical to EACMV-[TZ] and named EACMV-[TZ/YV]. One isolate of African cassava mosaic virus with 97% sequence identity with other isolates of ACMV was named ACMV-[TZ]. It represents the first ACMV isolate from Tanzania to be sequenced. The molecular variability of CMGs was also evaluated using partial B component nucleotide sequences of 13 EACMV isolates from Tanzania. Using the sequences of all CMGs currently available, we have shown the presence of a number of putative recombination fragments that are more prominent in all components of EACMV than in ACMV. This new knowledge about the molecular CMG diversity in East Africa, and in Tanzania in particular, has led us to hypothesize about the probable importance of this part of Africa as a source of diversity and evolutionary change both during the early stages of the relationship between CMGs and cassava and in more recent times. The existence of multiple CMG isolates with high DNA genome diversity in Tanzania and the molecular forces behind this diversity pose a threat to cassava production throughout the African continent. PMID- 15784146 TI - Do we need a Unique Scientist ID for publications in biomedicine? AB - BACKGROUND: The PubMed database contains nearly 15 million references from more than 4,800 biomedical journals. In general, authors of scientific articles are addressed by their last name and forename initial. DISCUSSION: In general, names can be too common and not unique enough to be search criteria. Today, Ph.D. students, other researchers and women publish scientific work. A person may not only have one name but several names and publish under each name. A Unique Scientist ID could help to address people in peer-to-peer (P2P) networks. As a starting point, perhaps PubMed could generate and manage such a scientist ID. SUMMARY: A Unique Scientist ID would improve knowledge management in science. Unfortunately in some of the publications, and then within the online databases, only one letter abbreviates the author's forename. A common name with only one initial could retrieve pertinent citations, but include many false drops (retrieval matching searched criteria but indisputably irrelevant). PMID- 15784147 TI - The excitement of Google Scholar, the worry of Google Print. AB - In late 2004 Google announced two major projects, the unveiling of Google Scholar and a major expansion of the Google Print digitization program. Both projects have generated discussion within the library and research communities, and Google Print has received significant media attention.This commentary describes exciting educational possibilities stimulated by Google Scholar, and argues for caution regarding the Google Print project. PMID- 15784148 TI - Efficacy of two distinct ethanol-based hand rubs for surgical hand disinfection - a controlled trial according to prEN 12791. AB - BACKGROUND: Aim of the study was to determine the efficacy of two distinct ethanol-based hand rubs for surgical hand disinfection in a controlled cross-over trial according to prEN 12791. METHODS: 20 subjects were included. Hands were washed for 1 min with soap. The bacterial prevalue was obtained by rubbing finger tips in TSB for 1 min. Then, each subject treated the hands with the reference procedure (n-propanol, 60% v/v) or the product (Sterillium Rub, based on 80% ethanol; Avagard, based on 61% ethanol and 1% chlorhexidine gluconate) which were all applied in 3 to 4 portions each of 3 ml for a total of 3 min. Bacterial postvalues (immediate effect) were taken from one hand, the other hand was gloved for 3 h. After gloves were taken off the second postvalue was taken for the assessment of a sustained effect. RESULTS: Bacterial pre-values were between 4.38 +/- 0.66 and 4.46 +/- 0.71. Sterillium Rub achieved the required immediate (mean log10-reduction of 2.59 +/- 1.19) and sustained effect (1.73 +/- 1.08) compared with the reference treatment (immediate effect: 2.58 +/- 1.16; sustained effect: 1.67 +/- 0.96). Avagard, however, did not achieve the required immediate (1.82 +/ 1.40) and sustained effect (1.41 +/- 1.08) in comparison to the reference disinfection (immediate effect: 2.98 +/- 0.90; sustained effect: 2.56 +/- 1.17; p < 0.01; Wilcoxon test). CONCLUSION: Based on our data, Sterillium Rub can be regarded to be effective for surgical hand disinfection, but Avagard can not. The addition of 1% chlorhexidine gluconate to 61% ethanol (w/w) did not outweigh an ethanol concentration of 80% (w/w). PMID- 15784149 TI - The antitumor immune response in HER-2 positive, metastatic breast cancer patients. AB - The aim of this study was to determine the basis for anti-tumor immune reactivity observed in patients with human epidermal growth factor receptor-2 (HER-2) (3+) breast carcinoma using an in vitro model in which the role of the HER-2-specific monoclonal antibody Herceptin was also investigated. Patients with metastatic breast cancer who had their primary tumor resected were included in this study. Peripheral blood mononuclear cell (PBMC)-dependent cytotoxicity in the presence or absence of Herceptin were assessed using the survival of target breast adenocarcinoma MDA-MB-361 cells as a parameter in a (3-(4,5-dimethyl-2-thiazolyl) 2,5-diphenyl-2H-tetrazolium bromide (MTT) test. We observed a significant increase in PBMC-dependent cytotoxicity when autologous serum was introduced in the assay. Furthermore, the addition of Herceptin significantly increases their cytotoxicity. These data suggest that autologous serum constitutively contains factors that might affect PBMC-dependent cytotoxic activity against HER-2 positive cancer cells. PMID- 15784150 TI - Do Indonesian medical practitioners approve the availability of emergency contraception over-the-counter? A survey of general practitioners and obstetricians in Jakarta. AB - BACKGROUND: Few studies have examined the attitude of medical practitioners towards the availability of emergency contraception (EC) without prescription. In Indonesia, EC (either Yuzpe regimen or Postinor-2) is available by prescription only. We aimed to examine the level of knowledge, attitudes and practices of medical practitioners in Indonesia about EC, in particular their attitudes to the availability of EC over-the-counter (OTC), using a questionnaire. METHODS: Data were collected by an anonymous structured questionnaire. Questionnaires were distributed to general practitioners in 36 Community Health Centres and 25 private clinics using stratified random sampling according to area in Jakarta, and to obstetricians practicising in 24 government and private hospitals and eight private clinics in Jakarta. Two hundred and five general practitioners and 142 obstetricians and gynaecologists participated; overall response rate was 75%. RESULTS: Although most participants were familiar with EC, only 22% received a very good knowledge score (4 or 5/5 answers correct), while 52% received a poor score (0-2/5 correct). Most participants did not support the OTC availability of EC (70%). Logistic regression identified that participants who prescribed EC had an Odds of 3.8 (95% CI 1.90, 7.73) of approving OTC EC, after adjustment for age and speciality. CONCLUSION: Although many organisations are working towards OTC availability of EC, it needs to be recognized and addressed that doctors who do not prescribe EC are unlikely to support the increased availability of EC. PMID- 15784151 TI - Global, local and focused geographic clustering for case-control data with residential histories. AB - BACKGROUND: This paper introduces a new approach for evaluating clustering in case-control data that accounts for residential histories. Although many statistics have been proposed for assessing local, focused and global clustering in health outcomes, few, if any, exist for evaluating clusters when individuals are mobile. METHODS: Local, global and focused tests for residential histories are developed based on sets of matrices of nearest neighbor relationships that reflect the changing topology of cases and controls. Exposure traces are defined that account for the latency between exposure and disease manifestation, and that use exposure windows whose duration may vary. Several of the methods so derived are applied to evaluate clustering of residential histories in a case-control study of bladder cancer in south eastern Michigan. These data are still being collected and the analysis is conducted for demonstration purposes only. RESULTS: Statistically significant clustering of residential histories of cases was found but is likely due to delayed reporting of cases by one of the hospitals participating in the study. CONCLUSION: Data with residential histories are preferable when causative exposures and disease latencies occur on a long enough time span that human mobility matters. To analyze such data, methods are needed that take residential histories into account. PMID- 15784152 TI - "Harshlighting" small blemishes on microarrays. AB - BACKGROUND: Microscopists are familiar with many blemishes that fluorescence images can have due to dust and debris, glass flaws, uneven distribution of fluids or surface coatings, etc. Microarray scans show similar artefacts, which affect the analysis, particularly when one tries to detect subtle changes. However, most blemishes are hard to find by the unaided eye, particularly in high density oligonucleotide arrays (HDONAs). RESULTS: We present a method that harnesses the statistical power provided by having several HDONAs available, which are obtained under similar conditions except for the experimental factor. This method "harshlights" blemishes and renders them evident. We find empirically that about 25% of our chips are blemished, and we analyze the impact of masking them on screening for differentially expressed genes. CONCLUSION: Experiments attempting to assess subtle expression changes should be carefully screened for blemishes on the chips. The proposed method provides investigators with a novel robust approach to improve the sensitivity of microarray analyses. By utilizing topological information to identify and mask blemishes prior to model based analyses, the method prevents artefacts from confounding the process of background correction, normalization, and summarization. PMID- 15784153 TI - Protein encoding genes in an ancient plant: analysis of codon usage, retained genes and splice sites in a moss, Physcomitrella patens. AB - BACKGROUND: The moss Physcomitrella patens is an emerging plant model system due to its high rate of homologous recombination, haploidy, simple body plan, physiological properties as well as phylogenetic position. Available EST data was clustered and assembled, and provided the basis for a genome-wide analysis of protein encoding genes. RESULTS: We have clustered and assembled Physcomitrella patens EST and CDS data in order to represent the transcriptome of this non-seed plant. Clustering of the publicly available data and subsequent prediction resulted in a total of 19,081 non-redundant ORF. Of these putative transcripts, approximately 30% have a homolog in both rice and Arabidopsis transcriptome. More than 130 transcripts are not present in seed plants but can be found in other kingdoms. These potential "retained genes" might have been lost during seed plant evolution. Functional annotation of these genes reveals unequal distribution among taxonomic groups and intriguing putative functions such as cytotoxicity and nucleic acid repair. Whereas introns in the moss are larger on average than in the seed plant Arabidopsis thaliana, position and amount of introns are approximately the same. Contrary to Arabidopsis, where CDS contain on average 44% G/C, in Physcomitrella the average G/C content is 50%. Interestingly, moss orthologs of Arabidopsis genes show a significant drift of codon fraction usage, towards the seed plant. While averaged codon bias is the same in Physcomitrella and Arabidopsis, the distribution pattern is different, with 15% of moss genes being unbiased. Species-specific, sensitive and selective splice site prediction for Physcomitrella has been developed using a dataset of 368 donor and acceptor sites, utilizing a support vector machine. The prediction accuracy is better than those achieved with tools trained on Arabidopsis data. CONCLUSION: Analysis of the moss transcriptome displays differences in gene structure, codon and splice site usage in comparison with the seed plant Arabidopsis. Putative retained genes exhibit possible functions that might explain the peculiar physiological properties of mosses. Both the transcriptome representation (including a BLAST and retrieval service) and splice site prediction have been made available on http://www.cosmoss.org, setting the basis for assembly and annotation of the Physcomitrella genome, of which draft shotgun sequences will become available in 2005. PMID- 15784154 TI - The incidence of prescribing errors in an eye hospital. AB - BACKGROUND: Relatively little is known about the incidence of prescribing errors and there has been no work on this in a single specialty ophthalmic hospital. Knowing where and when errors are most likely to occur is generally felt to be the first step in trying to prevent these errors. This study is an attempt in, the setting of an eye hospital, to try to identify and attribute these medication errors. METHODS: The study setting was a single specialty eye hospital geographically separated from the main general hospital. Pharmacists prospectively recorded the number of errors of prescribing during a 4 week period at an eye hospital in UK. The errors were categorised as error of prescription writing or drug error. Potential significance of the errors was not addressed. RESULTS: Overall 144/1952 (8%) prescription sheets had errors. 7% of the total errors were errors of prescription writing while 1% were drug errors. The majority of errors were made by junior doctors and no drug errors were made by senior doctors. The outpatients department had by far the highest prevalence of errors. CONCLUSION: Certain areas within the hospital and certain grades of staff are more prone to drug errors. Further study is required to look at the reasons why this is so and what systems can be put in place to reduce these errors. PMID- 15784155 TI - Risk-based early prevention in comparison with routine prevention of dental caries: a 7-year follow-up of a controlled clinical trial; clinical and economic aspects. AB - BACKGROUND: The results in an earlier study with 2-5-year-old children indicated that, in comparison with conventional prevention, a risk-based prevention programme was effective in reducing dental caries in a low-caries community. The aim of the present study was to examine the clinical and economic findings seven years after the cessation of the targeted programme, from the perspective of public health care. METHODS: The present material was collected from the dental records of the public health care centres, and included all dental visits after the 5-year examination until the 12-year examination. The groups were compared in relation to clinically detected caries at the age of 12 years, the number of dental visits needed from 5 to 12 years of age, and the estimation of running costs during these years. Statistical analyses included univariate analysis of variance, and calculation of absolute risk reduction and number needed to treat (NNT) values. RESULTS: At the age of 12 years, DMF was significantly related to the risk category determined ten years earlier, in both study groups. In the risk based group, the absolute risk reduction for caries in permanent dentition was 0.13 (95% confidence interval 0.06 - 0.21), and the associated NNT value was 8 (95% confidence interval 5 - 17). The total number of preventive, as well as restorative visits was lower in the risk-based than in the routine prevention group. The findings indicate that early risk-based prevention can be correctly targeted, clinically effective, and economically profitable also from the long term point of view. CONCLUSION: Early prevention of dental caries also has long term benefits in a 7-year follow-up perspective. This seems to hold true as regards targeting, as well as clinical and economic effectiveness. Success in risk-based prevention enables successful work division, and consequently, economic effectiveness. PMID- 15784157 TI - A meta-analysis of neuropsychological change to clozapine, olanzapine, quetiapine, and risperidone in schizophrenia. AB - Cognitive impairment is a core feature of schizophrenia and a major impediment to social and vocational rehabilitation. A number of studies have claimed cognitive benefits from treatment with various atypical antipsychotic drugs (APDs). The currently available evidence supporting cognitive improvement with atypical APDs was evaluated in two meta-analyses. Studies that (1) prospectively examined cognitive change to the atypical APDs clozapine, olanzapine, quetiapine, and risperidone, (2) included a commonly used neuropsychological test, and (3) provided data from which relevant effect sizes could be calculated, were included. Forty-one studies met these criteria. Neuropsychological test data from each study were combined into a Global Cognitive Index and nine cognitive domain scores. Two meta-analyses were carried out. The first included 14 controlled, random assignment trials that assigned subjects to an atypical APD and a typical APD control arm. The second analysis included all prospective investigations of atypical treatment and the within-group change score divided by its standard deviation served as an estimate of effect size (ES). The first analysis revealed that atypicals are superior to typicals at improving overall cognitive function (ES=0.24). Specific improvements were observed in the learning and processing speed domains. The second analysis extended the improvements to a broader range of cognitive domains (ES range=0.17-0.46) and identified significant differences between treatments in attention and verbal fluency. Moderator variables such as study blind and random assignment influence results of cognitive change to atypical APDs. Atypical antipsychotics produce a mild remediation of cognitive deficits in schizophrenia, and specific atypicals have differential effects within certain cognitive domains. PMID- 15784156 TI - Mutations of PIK3CA in gastric adenocarcinoma. AB - BACKGROUND: Activation of the phosphatidylinositol 3-kinase (PI3K) through mutational inactivation of PTEN tumour suppressor gene is common in diverse cancer types, but rarely reported in gastric cancer. Recently, mutations in PIK3CA, which encodes the p110alpha catalytic subunit of PI3K, have been identified in various human cancers, including 3 of 12 gastric cancers. Eighty percent of these reported mutations clustered within 2 regions involving the helical and kinase domains. In vitro study on one of the "hot-spot" mutants has demonstrated it as an activating mutation. METHODS: Based on these data, we initiated PIK3CA mutation screening in 94 human gastric cancers by direct sequencing of the gene regions in which 80% of all the known PIK3CA mutations were found. We also examined PIK3CA expression level by extracting data from the previous large-scale gene expression profiling study. Using Significance Analysis of Microarrays (SAM), we further searched for genes that show correlating expression with PIK3CA. RESULTS: We have identified PIK3CA mutations in 4 cases (4.3%), all involving the previously reported hotspots. Among these 4 cases, 3 tumours demonstrated microsatellite instability and 2 tumours harboured concurrent KRAS mutation. Data extracted from microarray studies showed an increased expression of PIK3CA in gastric cancers when compared with the non neoplastic gastric mucosae (p < 0.001). SAM further identified 2910 genes whose expression levels were positively associated with that of PIK3CA. CONCLUSION: Our data suggested that activation of the PI3K signalling pathway in gastric cancer may be achieved through up-regulation or mutation of PIK3CA, in which the latter may be a consequence of mismatch repair deficiency. PMID- 15784158 TI - Neuropeptide Y and corticotropin-releasing hormone in CSF mark response to antidepressive treatment with citalopram. AB - Neuropeptides appear to play a role in the pathophysiology of depression and electroconvulsive treatment and lithium affect these compounds in human cerebrospinal fluid (CSF) and rodent brain. Consequently, we investigated whether long-term treatment with the selective serotonin reuptake inhibitor (SSRI) citalopram (Cit) would also affect neuropeptides in CSF of depressed patients. Changes in CSF monoamine metabolites were also explored. CSF concentrations of corticotropin-releasing hormone (CRH)-like immunoreactivity (-LI), neuropeptide Y (NPY)-LI, and Cit were determined in 21 patients with major depression. Lumbar puncture was performed in the morning at baseline and was repeated after at least 4 wk of Cit treatment (40 mg/d). The severity of depression was assessed by the Hamilton Rating Scale for Depression (HAMD). Cit treatment was associated with a significant increase in NPY-LI and decrease in CRH-LI. An evaluation of the relationship between changes in concentrations of NPY-LI, CRH-LI, and the clinical response showed significant correlations between these parameters. Significant NPY and CRH changes in CSF following treatment as well as correlations to changes in HAMD support the hypothesis that these two peptides play a role in affective disorders and are markers of therapeutic response. PMID- 15784160 TI - Defensins and cathelicidins: neutrophil peptides with roles in inflammation, hyperlipidemia and atherosclerosis. AB - Atherosclerosis is a disease that begins in fetal life and represents a leading cause of morbidity and mortality associated with significant socioeconomic consequences. A central concept with regard to its pathogenesis is that of endothelial cell dysfunction, which is associated with the release of a large number of mediators secreted by leukocytes that are present in large numbers at the sites of atheroma formation. Neutrophil peptides defensins and cathelicidins are essential elements of the innate immunity and have been present in high concentrations in atherosclerotic plaques in humans. Recently, their role as potential mediators of vascular disease was investigated. Defensins are involved in the lipoprotein metabolism in the vessel wall, favoring LDL and lipoprotein (a) accumulation and modification in the endothelium and the extracellular matrix. They also interfere with the vascular smooth muscle cell function, exhibit prothrombotic activity, and play an inhibitory role in various phases of angiogenesis. Cathelicidins were recently found to enhance endothelial proliferation in cultures, induce functionally significant angiogenesis in animal models, and regulate endothelial cell apoptosis. Further study of these peptides could provide useful insight in the relationship between inflammation and atherosclerosis and is anticipated to have therapeutic and prognostic ramifications. PMID- 15784161 TI - Intestinal stem cells. AB - The intestinal tract has a rapid epithelial cell turnover, which continues throughout life. The process is regulated and maintained by a population of stem cells, which give rise to all the intestinal epithelial cell lineages. Studies in both the mouse and the human show that these cells are capable of forming clonal crypt populations. Stem cells remain hard to identify, however it is thought that they reside in a 'niche' towards the base of the crypt and their activity is regulated by the paracrine secretion of growth factors and cytokines from surrounding mesenchymal cells. Stem cell division is usually asymmetric with the formation of an identical daughter stem cell and committed progenitor cells. Progenitor cells retain the ability to divide until they terminally differentiate. Occasional symmetric division produces either 2 daughter cells with stem cell loss, or 2 stem cells and eventual clone dominance. This stochastic extinction of stem cell lines with eventual dominance of one cell line is called 'niche succession'. The discovery of plasticity, the ability of stem cells to engraft into, and in some cases replace the function of damaged host tissues has generated a large amount of scientific and clinical interest: however the concept remains controversial and is still a subject of hot debate. Studies are beginning to identify the complex molecular, genetic and cellular pathways underlying stem cell function such as Wnt signalling, bone morphogenetic protein (BMP) and Notch/Delta pathways. The derangement of these pathways within stem cells plays an integral part in the development of malignancy within the intestinal tract. PMID- 15784162 TI - The role of stem cells in cardiac regeneration. AB - After myocardial infarction, injured cardiomyocytes are replaced by fibrotic tissue promoting the development of heart failure. Cell transplantation has emerged as a potential therapy and stem cells may be an important and powerful cellular source. Embryonic stem cells can differentiate into true cardiomyocytes, making them in principle an unlimited source of transplantable cells for cardiac repair, although immunological and ethical constraints exist. Somatic stem cells are an attractive option to explore for transplantation as they are autologous, but their differentiation potential is more restricted than embryonic stem cells. Currently, the major sources of somatic cells used for basic research and in clinical trials originate from the bone marrow. The differentiation capacity of different populations of bone marrow-derived stem cells into cardiomyocytes has been studied intensively. The results are rather confusing and difficult to compare, since different isolation and identification methods have been used to determine the cell population studied. To date, only mesenchymal stem cells seem to form cardiomyocytes, and only a small percentage of this population will do so in vitro or in vivo. A newly identified cell population isolated from cardiac tissue, called cardiac progenitor cells, holds great potential for cardiac regeneration. Here we discuss the potential of the different cell populations and their usefulness in stem cell based therapy to repair the damaged heart. PMID- 15784163 TI - Transplantation of hematopoietic stem cells from the peripheral blood. AB - Hematopoietic stem cells can be collected from the peripheral blood. These hematopoietic stem cells (HSC), or better progenitor cells, are mostly expressed as the percentage of cells than react with CD34 antibodies or that form colonies in semi-solid medium (CFU-GM). Under steady-state conditions the number of HSC is much lower in peripheral blood than in bone marrow. Mobilization with chemotherapy and/or growth factors may lead to a concentration of HSC in the peripheral blood that equals or exceeds the concentration in bone marrow. Transplantation of HSC from the peripheral blood results in faster hematologic recovery than HSC from bone marrow. This decreases the risk of infection and the need for blood-product support. For autologous stem-cell transplantation (SCT), the use of peripheral blood cells has completely replaced the use of bone marrow. For allogeneic SCT, on the other hand, the situation is more complex. Since peripheral blood contains more T-lymphocytes than bone marrow, the use of HSC from the peripheral blood increases the risk of graft-versus-host disease after allogeneic SCT. For patients with goodrisk leukemia, bone marrow is still preferred, but for patients with high-risk disease, peripheral blood SCT has become the therapy of choice. PMID- 15784164 TI - HSP60, Bax, apoptosis and the heart. AB - HSP60 has primarily been known as a mitochondrial protein that is important for folding key proteins after import into the mitochondria. It is now clear that a significant amount of HSP60 is also present in the extra-mitochondrial cytosol of many cells. In the heart, this cytosolic HSP60 complexes with Bax, Bak and Bcl XL, but not with Bcl-2. Reduction in HSP60 expression precipitates apoptosis, but does not alter mitochondrial function. During hypoxia, HSP60 cellular distribution changes, with HSP60 leaving the cytosol, and translocating to the plasma membrane. Total cellular HSP60 does not change until 10 h of reoxygenation; however, release of cytochrome c from the mitochondria occurs prior to reoxygenation, coinciding with the redistribution of HSP60. The changes in HSP60, Bax and cytochrome c during hypoxia can be replicated by ATP depletion. HSP60 has also been shown to accelerate the cleavage of pro-caspase3. Thus, HSP60 has a complex role in apoptosis in the cell. Its binding to Bax under normal conditions suggests a key regulatory role in apoptosis. PMID- 15784165 TI - The activation of Akt/PKB signaling pathway and cell survival. AB - Akt/PKB is a serine/threonine protein kinase that functions as a critical regulator of cell survival and proliferation. Akt/PKB family comprises three highly homologous members known as PKBalpha/Akt1, PKBbeta/Akt2 and PKBgamma/Akt3 in mammalian cells. Similar to many other protein kinases, Akt/PKB contains a conserved domain structure including a specific PH domain, a central kinase domain and a carboxyl-terminal regulatory domain that mediates the interaction between signaling molecules. Akt/PKB plays important roles in the signaling pathways in response to growth factors and other extracellular stimuli to regulate several cellular functions including nutrient metabolism, cell growth, apoptosis and survival. This review surveys recent developments in understanding the molecular mechanisms of Akt/PKB activation and its roles in cell survival in normal and cancer cells. PMID- 15784166 TI - Engineering bone: challenges and obstacles. AB - Repair of large bone defects is still a challenge for the orthopaedic, reconstructive and maxillo-facial surgeon. Availability of pluripotent stem cells from either autologous or allogenic sources and the potential of inducing the osteogenic phenotype is motivating exploration and development of custom-tailored materials known as "bioengineered bone constructs". In such cases, the clinical scenario involves either expansion of stem cells in monolayer and loading them into a porous scaffold prior to surgery or direct cell expansion within the scaffold, and implanting this novel construct back into the donor patient. In this review, we delineate, from an engineering perspective, the progress that has been made to date and the challenges remaining in successfully translating this promising (but not yet definitively established) approach from bench to the bed site. PMID- 15784167 TI - Gene activation and protein expression following ischaemic stroke: strategies towards neuroprotection. AB - Current understanding of the patho-physiological events that follow acute ischaemic stroke suggests that treatment regimens could be improved by manipulation of gene transcription and protein activation, especially in the penumbra region adjacent to the infarct. An immediate reduction in excitotoxicity in response to hypoxia, as well as the subsequent inflammatory response, and beneficial control of reperfusion via collateral revascularization near the ischaemic border, together with greater control over apoptotic cell death, could improve neuronal survival and ultimately patient recovery. Highly significant differences in gene activation between animal models for stroke by middle cerebral artery occlusion, and stroke in patients, may explain why current treatment strategies based on animal models of stroke often fail. We have highlighted the complexities of cellular regulation and demonstrated a requirement for detailed studies examining cell specific protective mechanisms after stroke in humans. PMID- 15784168 TI - Important roles of reversible acetylation in the function of hematopoietic transcription factors. AB - Hematopoiesis is a very complex process whose proper functioning requires the regulated action of a number of transcription factors. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) play significant roles in the regulation of hematopoietic transcription factors activity. Transcription factors such as GATA-1, EKLF, NF-E2, GATA-1, PU.1 recruit HATs and HDACs to chromatin, leading to histone acetylation and deacetylation, that affect chromatin structure and result in gene expression changes. On the other hand, transcription factors themselves can be acetylated and deacetylated by HATs and HDACs, respectively. Consequently, some important functions of these transcription factors are influenced, including DNA binding, transcription activation, repressor activity and proteinprotein interactions. The regulation of hematopoietic transcription factors activity by HATs and HDACs may serve as a good model for studying how tissue-specific and lineage-specific gene expression is controlled through acetylation/ deacetylation of histone/nonhistone proteins. PMID- 15784169 TI - Preferential activity of Tie2 promoter in arteriolar endothelium. AB - The tyrosine kinase Tie2/Tek (the receptor for angiopoietins) is considered one of the most reliable markers of the endothelial phenotype, across organisms, organs, and developmental stages. However, endothelium is intrinsically heterogeneous in origin, composition and function, presenting an arteriolar/venular asymmetry. In this regard, the expression of Tie2 along the vascular tree, although thought to be homogenous, has not been systematically investigated. Therefore we questioned whether the activity of Tie2 promoter is uniform in the microvascular endothelium. To this end, we analyzed in situ the expression of the markers beta-galactosidase [LacZ(Tie2)] and green fluorescent protein (GFP) [GFP(Tie2)], placed under the Tie2 promoter in transgenic mice, in whole mount tissue samples, which allow the simultaneous evaluation of its relative distribution in various microvascular compartments. In the mesenteries of LacZ(Tie2) and GFP(Tie2) mice, we found that the activity of Tie2 promoter is asymmetrically distributed, being much stronger in arteries and arterioles than on the venular side of the vascular tree. This observation was replicated in the diaphragm of LacZ(Tie2) mice. The capillaries presented a mosaic pattern of Tie2 promoter activity. Stimulation of angiogenesis either by wounding, or by intraperitoneal injection of Vascular Endothelial Growth Factor (VEGF), revealed that the arteriolar/venular asymmetry is established at endothelial cellular level early during new capillary formation, even before the starting of the microvascular blood flow. In conclusion, a strong Tie2 promoter activity qualifies as a novel marker of the arteriolar phenotype in microvascular endothelium. PMID- 15784170 TI - Expression of scaffolding, signalling and contractile-filament proteins in human myometria: effects of pregnancy and labour. AB - Successful parturition requires the co-ordination of numerous myometrial signalling events to allow for timely and efficient uterine contractions. Late pregnancy and labour onset in humans may be associated with changes in the expression of myometrial proteins implicated in such uterine contractile signal integration. Accordingly, in myometria from non-pregnant women and pregnant women, not in labour or in labour, we examined the content of putative plasmalemmal scaffolding proteins (caveolin-1 and -2) and compared these to the proportions of signal transducing rho-associated kinases (ROKalpha and beta) and contractile filament-associated proteins alpha-actin, myosin regulatory light chain (MLC(20)) and h-caldesmon. There was no effect of pregnancy or labour on the proportion of caveolin, ROK betaor alpha-actin. However, pregnancy was associated with a decrease in ROKalpha and MLC(20) such that ROK alpha: alpha actin and MLC(20): alpha-actin ratios were reduced compared to myometria of non pregnant women. In contrast, h-caldesmon was up-regulated in pregnancy resulting in an elevated h-caldesmon: alpha-actin ratio. There were, however, no further significant changes in ROK alpha, MLC(20) or h-caldesmon expression with spontaneous or oxytocin-induced labour. These data suggest that the mechanism(s) integrating myometrial signalling events with the onset of human labour does not involve differential alterations of the cellular expressions of caveolins, ROK, alpha-actin, MLC(20) or h-caldesmon. PMID- 15784171 TI - Relationship of eNOS gene variants to diseases that have in common an endothelial cell dysfunction. AB - The endothelial cell (EC) dysfunction is a common characteristic of various pathologies that include atherosclerosis, hypertension, and Fabry's disease. Aware of the role of eNO and ACE in EC dysfunction, we questioned whether polymorphism of eNOS and/or ACE gene may be a common denominator in these pathologies. Patients with CHD (108), HT (109), Fabry's disease (37) and healthy subjects (control, 141) were genotyped for the eNOSG894T by RFLP-PCR technique and for eNOS4b/a, and ACEI/D polymorphisms by PCR amplification. The results of these studies were statistically evaluated. Compared to controls, the frequency of the eNOSG894T (T allele) was higher in CHD (P=0.03) and Fabry (P=0.01), while the eNOS4b/a (a allele) in CHD (P=0.01) and HT patients (P=0.01). The proportion of the ACEI/D was similar in all subjects. In CHD patients at "low risk" of atherogenic factors, the frequency of the T and a alleles of eNOS gene was high (P=0.03 and 0.02, respectively). Carriers of the T allele of eNOSG894T were over represented (P=0.04) in Fabry subgroup with renal failure. Compared to women, the eNOS894T alleles were more frequent (P=0.03) in men with CHD and HT, whereas ACE I/D in men (P=0.03) with HT. These findings suggest: (i) the frequency of eNOSG894T and/or eNOS4b/a is significantly associated with coronary dysfunction; (ii) eNOS4b/a confers a relatively high risk of hypertension in subjects with atherogenic risk factors; (iii) the frequency of eNOSG894T is high in Fabry hemizygotes with renal complications. Therefore, eNOS gene polymorphism represent a frequent risk factor for vascular abnormalities in CHD, HT and Fabry's disease, afflictions which have in common, the endothelial dysfunction. PMID- 15784172 TI - The predictive value of peritubular capillaries C3d deposition in IgA glomerulonephritis. AB - The IgA glomerulonephritis (IgAGN) is one of the most common primary glomerulonephritis and has a variable and difficult to predict evolution toward the end-stage nephrosclerosis. The deposition of C3d complement component in peritubular capillaries (PTCs) indicates a variant type of acute rejection while C3d deposition in primary glomerulonephritis (GN) is poorly documented. The aim of this study is to examine C3d expression in peritubular capillaries (PTCs) as a predictive marker and its correlation with the severity of renal injury in IgA glomerulonephritis. Polyclonal FITC conjugated rabbit anti-human C3c and C3d antibodies were used for direct immunofluorescent evaluation of the C3c and C3d deposits in 24 kidney biopsies with IgA glomerulonephritis. The study revealed that the C3d deposits in peritubular capillaries were associated with known predictors for rapid progression of IgAGN: glomerular sclerosis (63.6%), atrophic tubules (90.9%) and interstitial sclerosis (81.8%). The intensity of the C3c glomerular immunofluorescent deposits was related with active lesions. Thus, the predictive value of C3d deposition on PTCs in IgAGN is worth to be taken into consideration as an unfavorable outcome of the disease and request further long run investigations. PMID- 15784173 TI - Genomic rearrangements on VCAM1, SELE, APEG1and AIF1 loci in atherosclerosis. AB - The inflammatory nature of atherosclerosis has been well established. However, the initial steps that trigger this response in the arterial intima remain obscure. Previous studies reported a significant rate of genomic alterations in human atheromas. The accumulation of genomic rearrangements in vascular endothelium and smooth muscle cells may be important for disease development. To address this issue, 78 post-mortem obtained aortic atheromas were screened for microsatellite DNA alterations versus correspondent venous blood. To evaluate the significance of these observations, 33 additional histologically normal aortic specimens from age and sex-matched cases were examined. Loss of heterozygosity (LOH) was found in 47,4% of the cases and in 18,2% of controls in at least one locus. The LOH occurrence in aortic tissue is associated to atherosclerosis risk (OR 4,06, 95% CI 1,50 to 10,93). Significant genomic alterations were found on 1p32-p31, 1q22-q25, 2q35 and 6p21.3 where VCAM1, SELE, APEG1 and AIF1 genes have been mapped respectively. Our data implicate somatic DNA rearrangements, on loci associated to leukocyte adhesion, vascular smooth muscle cells growth, differentiation and migration, to atherosclerosis development as an inflammatory condition. PMID- 15784174 TI - Computerized morphometric discrimination between normal and tumoral cells in oral smears. AB - The oral exfoliative cytology allows a quick and fairly accurate assessment of suspicious lesions of the oral cavity. Within this context, our paper proposes a quantitative approach, focusing on the construction of a classifier for detecting the presence of the tumoral cells on oral smears. The design of the classifier relies on a detailed computerized analysis of the individual morphometric features exhibited by two large known populations of normal and tumoral cells, respectively; the digital image processing was performed in the Zeiss KS400 environment. The classifier was implemented as a neural network with step activation function, whose parameters were obtained from an adequate training, based on the nuclear and cytoplasmic areas of the cells belonging to the two populations. Our procedure based on this classifier was meant to operate by identifying the tumoral or normal nature of any cell randomly selected from a smear. To identify the nature of an arbitrary cell, its nuclear and cytoplasmic areas are presented at the input of the classifier. The classification procedure was tested on several smears, and the results coincided with the pathological diagnosis in all the considered cases. The performances of our approach are discussed in comparison with other analytical methods previously reported in oral exfoliative cytology. These discussions emphasize the role of numerical information exploited for the classifier design, concluding that the individual morphometric features are more meaningful than the global characterization of smears by mean values. PMID- 15784176 TI - Potential applications of intracellular antibodies (intrabodies) in stem cell therapeutics. PMID- 15784175 TI - Interstitial cells of Cajal in pancreas. AB - We show here (presumably for the first time) a special type of cell in the human and rat exocrine pancreas. These cells have phenotypic characteristics of the enteric interstitial cells of Cajal (ICC). To identify pancreatic interstitial cells of Cajal (pICC) we used routine light microscopy, non-conventional light microscopy (less than 1 mum semi-thin sections of Epon-embedded specimens cut by ultramicrotomy and stained with Toluidine blue), transmission electron microscopy (TEM), and immunocytochemistry. The results showed that pICC can be recognized easily by light microscopy, particularly on semi-thin sections, as well as by TEM. Two-dimensional reconstructions from serial photos suggest a network-like spatial distribution of pICC. pICC represent 3.3+/-0.5% of all pancreatic cells, and seem to establish close spatial relationships with: capillaries (43%), acini (40%), stellate cells (14%), nerve fibres (3%). Most of pICC (88%) have 2 or 3 long processes (tens of mum) emerging from the cell body. TEM data show that pICC meet the criteria for positive diagnosis as ICC (e.g. numerous mitochondria, 8.7+/-0.8% of cytoplasm). Immunocytochemistry revealed that pICC are CD117/c-kit and CD34 positive. We found pICC positive (40-50%) for smooth muscle alpha-actin or S-100, and, occasionally, for CD68, NK1 neurokinin receptor and vimentin. The reactions for desmin and chromogranin A were negative in pICC. At present, only hypotheses and speculations can be formulated on the possible role of the pICC (e.g., juxtacrine and/or paracrine roles). In conclusion, the quite-established dogma: "ICC only in cavitary organs" is overpassed. PMID- 15784177 TI - A concept on the role of Helicobacter pylori infection in autoimmune pancreatitis. AB - Autoimmune pancreatitis, an inflammatory process of the pancreas due to an autoimmune mechanism establishing etiology of chronic pancreatitis, is characterized by the presence of autoantibodies, hypergammaglobulinemia, pancreatic enlargement, pancreatic duct strictures, and pathologic features of fibrotic changes with intense, mainly lymphocytic infiltrations, which may contribute to tissue destruction probably by apoptosis. In almost 60% of the cases, this type of pancreatitis coexists with other autoimmune diseases such as Sjogren's syndrome, sclerosing extrahepatic cholangitis, primary biliary cirrhosis, autoimmune hepatitis, or other extrapancreatic disorders, and recently with gastric peptic ulceration. The diversity of extrapancreatic lesions with similar histopathologic findings suggests general involvement of the digestive system in this disease, although the presence of such involvement has not been fully elucidated. Similarly, Helicobacter pylori (H. pylori) infection, a well known cause of gastric ulcer, has been associated, via molecular mimicry of host structures by its constituents with the same autoimmune conditions, also characterized by fibrotic changes and/or lymphoplasmacytic inflammations, accompanied by aberrations of T cell apoptosis that contribute to hepatobiliary- or extrahepatic-tissue destruction. Considering that H. pylori is involved in the pathogenesis and pathophysiology of these autoimmune disorders, we propose that this organism might trigger autoimmune pancreatitis through induction of autoimmunity and apoptosis. PMID- 15784178 TI - Understanding breast cancer risk -- where do we stand in 2005? AB - Breast cancer is the most frequent cancer in women and represents the second leading cause of cancer death among women (after lung cancer). The etiology of breast cancer is still poorly understood with known breast cancer risk factors explaining only a small proportion of cases. Risk factors that modulate the development of breast cancer discussed in this review include: age, geographic location (country of origin) and socioeconomic status, reproductive events, exogenous hormones, lifestyle risk factors (alcohol, diet, obesity and physical activity), familial history of breast cancer, mammographic density, history of benign breast disease, ionizing radiation, bone density, height, IGF- 1 and prolactin levels, chemopreventive agents. Additionally, we summarized breast cancer risk associated with the following genetic factors: breast cancer susceptibility high-penetrance genes (BRCA1, BRCA2, p53, PTEN, ATM, NBS1 or LKB1) and low-penetrance genes such as cytochrome P450 genes (CYP1A1, CYP2D6, CYP19), glutathione S-transferase family (GSTM1, GSTP1), alcohol and one-carbon metabolism genes (ADH1C and MTHFR), DNA repair genes (XRCC1, XRCC3, ERCC4/XPF) and genes encoding cell signaling molecules (PR, ER, TNFalpha or HSP70). All these factors contribute to a better understanding of breast cancer risk. Nonetheless, in order to evaluate more accurately the overall risk of breast tumorigenesis, novel genetic and phenotypic traits need to be identified. PMID- 15784179 TI - Purification and characterization of RihC, a xanthosine-inosine-uridine-adenosine preferring hydrolase from Salmonella enterica serovar Typhimurium. AB - Salmonella enterica serovar Typhimurium normally salvage nucleobases and nucleosides by the action of nucleoside phosphorylases and phosphoribosyltransferases. In contrast to Escherichia coli, which catabolizes xanthosine by xanthosine phosphorylase (xapA), Salmonella cannot grow on xanthosine as the sole carbon and energy source. By functional complementation, we have isolated a nucleoside hydrolase (rihC) that can complement a xapA deletion in E. coli and we have overexpressed, purified and characterized this hydrolase. RihC is a heat stable homotetrameric enzyme with a molecular weight of 135 kDa that can hydrolyze xanthosine, inosine, adenosine and uridine with similar catalytic efficiency (k(cat)/Km=1 to 4 x 10(4) M(-1)s(-1)). Cytidine and guanosine is hydrolyzed with approximately 10-fold lower efficiency (k(cat)/Km=0.7 to 1.2 x 10(3) M(-1)s(-1)) while RihC is unable to hydrolyze the deoxyribonucleosides thymidine and deoxyinosine. The Km for all nucleosides except adenosine is in the mM range. The pH optimum is different for inosine and xanthosine and the hydrolytic capacity (k(cat)/Km) is 5-fold higher for xanthosine than for inosine at pH 6.0 while they are similar at pH 7.2, indicating that RihC most likely prefers the neutral form of xanthosine. PMID- 15784180 TI - New GlcNAc/GalNAc-specific lectin from the ascidian Didemnum ternatanum. AB - Previously we isolated GlcNAc-specific lectin (DTL) from the ascidian Didemnum ternatanum by affinity chromatography on cross-linked ovalbumin. Here we report the purification and characterization of new D-GlcNAc/D-GalNAc-specific lectin DTL-A from the same ascidian. This lectin was isolated from non-bound cross linked ovalbumin fraction and further was purified by gel filtration on Sepharose CL-4B, affinity chromatography on GlcNAc-agarose and gel filtration on Superdex 200. SDS-polyacrylamide gel electrophoresis and gel filtration of purified lectin on Sepharose CL-4B indicates that it exists as large aggregates in the native state. Investigations of the carbohydrate specificity of DTL-A by enzyme-linked lectin assay suggest the multi-specificity of this lectin. DTL-A binds BSM, asialo-BSM as well as heparin and dextran sulfate. The binding of DTL-A to BSM was inhibited by monosaccharides D-GlcNAc and D-GalNAc, their alpha- but not beta anomers. Among polysaccharides and glycoconjugates, DTL-A binding to BSM was effectively inhibited by BSM, asialo-BSM, pronase-treated BSM and synthetic alpha D-GalNAc-PAA. Fetuin and asialofetuin showed a much lower inhibitory potency, heparin and dextran sulfate were noninhibitory. On the other hand, DTL-A binding to heparin was effectively inhibited by dextran sulfate, fucoidan, whereas BSM showed insignificantly inhibitory effect. DTL-A binding to heparin was not inhibited by D-GlcNAc and D-GalNAc. PMID- 15784181 TI - Genome-wide analysis reveals strong correlation between CpG islands with nearby transcription start sites of genes and their tissue specificity. AB - It has been envisaged that CpG islands are often observed near the transcriptional start sites (TSS) of housekeeping genes. However, neither the precise positions of CpG islands relative to TSS of genes nor the correlation between the presence of the CpG islands and the expression specificity of these genes is well-understood. Using thousands of sequences with known TSS in human and mouse, we found that there is a clear peak in the distribution of CpG islands around TSS in the genes of these two species. Thus, we classified human (mouse) genes into 6600 (2948) CpG+ genes and 2619 (1830) CpG- ones, based on the presence of a CpG island within the -100: +100 region. We estimated the degree of each gene being a housekeeper by the number of cDNA libraries where its ESTs were detected. Then, the tendency that a gene lacking CpG islands around its TSS is expressed with a higher degree of tissue specificity turned out to be evolutionarily conserved. We also confirmed this tendency by analyzing the gene ontology annotation of classified genes. Since no such clear correlation was found in the control data (mRNAs, pre-mRNAs, and chromosome banding pattern), we concluded that the effect of a CpG island near the TSS should be more important than the global GC content of the region where the gene resides. PMID- 15784182 TI - Specific human antibodies do not inhibit Trypanosoma cruzi oligopeptidase B and cathepsin B, and immunoglobulin G enhances the activity of trypomastigote secreted oligopeptidase B. AB - Trypanosoma cruzi expresses oligopeptidase B and cathepsin B that have important functions in the interaction with mammalian host cells. In this study, we demonstrated that sera from both chagasic rabbits and humans have specific antibodies to highly purified native oligopeptidase B and cathepsin B. Levels of antibodies to cathepsin B were higher than those observed to oligopeptidase B by absorbance values recorded upon ELISA. We next showed that 90% and 30% of sera from individuals with mucocutaneous leishmaniasis have antibodies that recognize oligopeptidase B and cathepsin B as antigens, respectively. In addition, 55% and 40% of sera from kala-azar patients have antibodies to oligopeptidase B and cathepsin B, respectively. Sera from malaria patients did not recognize the proteases as antigens. Despite high levels of specific antibodies, sera from T. cruzi-infected patients did not inhibit the activities of either oligopeptidase B or cathepsin B. Furthermore, sera or IgG purified from either infected or non infected individuals enhanced the enzymatic activity of the secreted oligopeptidase B. Oligopeptidase B secreted by trypomastigotes and cathepsin B released upon parasite lysis retain their enzymatic activities and may be associated with Chagas' disease pathogenesis by hydrolyzing host proteins and inducing host immune responses. PMID- 15784183 TI - Unique characteristics of HIV-1 Vif expression. AB - We examined the steady-state expression in cells of four accessory proteins of human immunodeficiency virus type 1 (HIV-1). For this purpose, a series of single gene expression vectors for these viral proteins were constructed and were monitored for their production by transfection. Among them, the expression level of Vif was found to be lowest in both the absence and presence of APOBEC3G. In addition, we noticed the presence of its truncated form, which was not observed for the other accessory proteins. When a subgenomic vector was used for transfection, authentic and several small forms of Vif were produced. By mutational analysis, these forms were demonstrated to be mutant Vif proteins translated from M8, M16 and M29. When a full-length molecular clone was used, the smaller versions of Vif were hardly observed. Functional analysis of these mutant Vif proteins showed that they are incapable of modulating viral infectivity. The results described above, i.e. the low steady-state expression and the presence of truncated forms, represent the unique characteristics of HIV-1 Vif. PMID- 15784184 TI - Th2 predominance and CD8+ memory T cell depletion in patients with severe acute respiratory syndrome. AB - The immune spectrum of severe acute respiratory syndrome (SARS) is poorly understood. To define the dynamics of the immune spectrum in SARS, serum levels of cytokines, chemokines, immunoglobulins, complement and specific antibodies against SARS-associated coronavirus (SARS-CoV) were assayed by enzyme-linked immunosorbent assay (ELISA), and phenotypes of peripheral lymphocytes were analyzed by flow cytometry in 95 SARS-infected patients. Results showed that interleukin (IL)-10 and transforming growth factor beta (TGF-beta) were continuously up-regulated during the entirety of SARS. Regulated on activation normally T cell-expressed and secreted (RANTES) levels were decreased, while monocyte chemoattractant protein-1 (MCP-1) was elevated in acute patients. Immunoglobulins and complement were elevated during the first month of SARS. Both serum-positive rates and titers of specific IgM and IgG antibodies responding to SARS-CoV peaked at days 41-60 from the onset of SARS. CD4+ and CD8+ T lymphocytes decreased significantly in acute-phase. CD3+CD8+CD45RO+ T lymphocytes were decreased by 36.78% in the convalescent patients. CONCLUSION: SARS-CoV seemed to elicit effective humoral immunity but inhibited cellular immunity, especially CD8+ memory T lymphocytes over time. Prolonged overproduction of IL-10 and TGF beta may play an important role in the disease. PMID- 15784185 TI - Immunoreactivity and differential developmental expression of known and putative Chlamydia trachomatis membrane proteins for biologically variant serovars representing distinct disease groups. AB - Chlamydia trachomatis is an intracellular bacterium that causes ocular and urogenital diseases worldwide. Membrane proteins have only been partially characterized, and the discovery of a nine-member polymorphic membrane protein gene family has enhanced interest in defining their function. We previously reported two putative insertion sequence-like elements in pmpC for biovariant Ba and one each for G and L2, suggesting horizontal gene transfer. Because of this and the tissue tropism differences for these biovariants, we analyzed by quantitative real-time RT-PCR pmpC expression relative to immunogenic protein genes ompA, groEL and gseA throughout development. Sera from infected adolescents were reacted by immunoblot against recombinant (r)PmpC and rMOMP. ompA and groEL revealed different developmental transcriptome profiles among the biovariants. pmpC expression occurred at 2 h, peaked at 18 for L2 (at 24 for Ba and G), with the highest mRNA levels throughout development for L2. pmpC expression as a function of time paralleled ompA expression with higher mRNA levels compared with groEL later in development. Only sera from D-, E- and G-infected patients reacted to rPmpC; all infected patients reacted to rMOMP. pmpC expression during logarithmic growth suggests a role in membrane building and/or integrity, which is supported by the presence of a signal peptidase and C-terminal phenylalanine in PmpC. Because phylogenetic analyses of pmpC segregate serovars according to tissue tropism, we speculate that biovariant transcriptome differences may contribute to this tropism. The heterogeneous biovariant pmpC expression throughout development and differential PmpC immunoreactivity also suggest a role for pmpC in antigenic variation. PMID- 15784186 TI - TGF-beta prevents eosinophilic lung disease but impairs pathogen clearance. AB - Respiratory infections are the third leading cause of death worldwide. Complications arise directly as a consequence of pathogen replication or indirectly due to aberrant or excessive immune responses. In the following report, we evaluate the efficacy, in a murine model, of nasally delivered DNA encoding TGF-beta1 to suppress immunopathology in response to a variety of infectious agents. A single nasal administration suppressed lymphocyte responses to Cryptococcus neoformans, influenza virus and respiratory syncytial virus. The suppression did not depend on the phenotype of the responding T cell, since both Th1 and Th2 responses were affected. During Th2-inducing infection, pulmonary eosinophilic responses were significantly suppressed. In all cases, however, suppressed immunity correlated with increased susceptibility to infection. We conclude that nasal TGF-beta treatment could be used to prevent pulmonary, pathogen-driven eosinophilic disease, although anti-pathogen strategies will need to be administered concordantly. PMID- 15784188 TI - Impact of anthropometric, lifestyle, and body composition variables on ultrasound measurements in school children. AB - Quantitative ultrasound (QUS) measurement at hand phalanges was demonstrated to be a reliable method to assess skeletal maturation during childhood and adolescence. The aim of the study was to evaluate the influence of age, gender, puberty, lifestyle factors, and body composition on QUS parameters and to provide a normative database for QUS in school children in Lebanon. Measurements of phalangeal osteosonography were examined in 256 healthy subjects (132 boys and 124 girls) aged 11-18 years using an ultrasound device. In both genders, amplitude-dependent speed of sound (AD-SoS) and bone transmission time (BTT) increased significantly with age and pubertal stages. Girls had higher AD-SoS values than boys between 11 and 15 years of age and at Tanner stages III and IV; however, no differences were detected in the older age groups. AD-SoS and BTT showed a significant positive correlation with age and height in both genders (R = 0.41-0.66, P < 0.01). There was no correlation between physical activity, calcium intake, sun exposure, and any of the QUS parameters in either gender. Weight showed moderate positive correlation with AD-SoS in boys and with BTT in both genders (R = 0.31-0.47, P < 0.01). Lean mass showed significant positive correlation with AD-SoS and BTT (R = 0.2-0.68, P < 0.01) in both genders. Percentage body fat showed significant negative correlation with BTT and AD-SoS in boys (R = -0.25 to -0.37, P < 0.01). In the linear regression analyses, there was a significant negative correlation between percentage fat mass and both AD SoS and BTT in both genders. In conclusion, QUS parameters of the phalanges in Lebanese children are related to growth variables such as height, age, and puberty in healthy children. The impact and magnitude of body composition variables and lifestyle factors on ultrasonometry derived variables differ from their effect on dual energy X-ray absorptiometry derived parameters. PMID- 15784187 TI - Dahl salt-sensitive rats develop hypovitaminosis D and hyperparathyroidism when fed a standard diet. AB - The Dahl salt-sensitive rat (S), a model for salt-sensitive hypertension, excretes protein-bound 25-hydroxyvitamin D (25-OHD) into urine when fed a low salt diet. Urinary 25-OHD increases during high salt intake. We tested the hypothesis that continuous loss of 25-OHD into urine would result in low plasma 25-OHD concentration in mature S rats raised on a standard diet. Dahl S and salt resistant (R) male rats were raised to maturity (12-month-old) on a commercial rat diet (1% salt) and switched to 0.3% (low) or 2% (high) salt diets 3 weeks before euthanasia. Urine (24 h) was collected at the end of the dietary treatments. Urinary 25-OHD and urinary 25-OHD binding activity of S rats were three times that of R rats, resulting in lower plasma 25-OHD and 24,25 dihydroxyvitamin D concentrations in S rats than in R rats (P < 0.001). Plasma parathyroid hormone concentrations of S rats were twice that of R rats. S rats fed 2% salt had higher plasma 1,25-dihydroxyvitamin D concentrations than those fed 0.3% salt (P = 0.002). S rats excreted more calcium into urine than R rats (P < 0.001) and did not exhibit the expected calciuric response to salt. Proteinuria of the S rats was three times that of the R rats, suggesting kidney damage in the S rats. Low plasma 25-OHD and 24,25-dihydroxyvitamin D and high plasma 1,25 dihydroxyvitamin D and PTH concentrations seen in the mature S rats have also been reported for elderly patients with low-renin (salt-induced) hypertension. An implication of this study is that low vitamin D status may occur with age in salt sensitive individuals, even when salt intake is normal. PMID- 15784189 TI - Bone and muscle protective potential of the prostate-sparing synthetic androgen 7alpha-methyl-19-nortestosterone: evidence from the aged orchidectomized male rat model. AB - This study reports the preclinical evaluation of the bone and muscle protective potential of the synthetic androgen 7alpha-methyl-19-nortestosterone (MENTtrade mark), as assessed in the aged orchidectomized rat model. Aged (13-month-old) orchidectomized Wistar rats were treated with different doses of MENT (4, 12 or 36 microg/day) subcutaneously for 16 weeks via mini-osmotic pumps. Analysis of the effects of androgen deficiency versus MENT replacement was performed using quantitative computed tomography (pQCT), dual energy X-ray absorptiometry (DEXA) and biochemical markers of bone turnover. At the end of the study period, prostate weight in orchidectomized rats treated with low- (4 microg/day) or mid dose (12 mug/day) MENT remained significantly lower compared to the sham-operated animals (-47% and -25%, respectively). High-dose MENT (36 microg/day), on the other hand, induced prostate hypertrophy (+21% versus sham). Low-, mid- and high dose MENT were found to be effective in suppressing the acceleration of bone remodeling following orchidectomy, as assessed by osteocalcin and deoxypyridinoline. In addition, low-, mid- and high-dose were able to prevent the orchidectomy-induced bone loss, as evaluated by DEXA at the femur and total-body and by pQCT at the femur. Compared to sham-operated animals, the low- and mid dose MENT groups showed no decline in lean body mass and no muscle atrophy (as measured by m. quadriceps weight) at 16 weeks, whereas high-dose MENT was associated with a significant decline in lean body mass (-8.5% versus sham) and quadriceps weight (-10.6%). We conclude that, in the aged orchidectomized rat model, low- and mid-doses of the synthetic androgen MENT have bone and muscle protective effects and do not induce prostate hypertrophy. The bone protective action of high-dose MENT, however, occurs at the expense of muscle wasting and prostate hypertrophy. Our findings support the need for human studies to explore the potential of MENT as an option for androgen replacement in aging men. PMID- 15784190 TI - Platelets can neutralize hydrogen peroxide in an acute toxicity model with cells involved in granulation tissue formation. AB - Platelets play a key role in the replacement of the blood clot with granulation tissue during the early steps of bone regeneration. We hypothesized that activated platelets can neutralize locally produced reactive oxygen species, thereby protecting cells involved in granulation tissue formation. The potential of platelet-released supernatant (PRS) to neutralize hydrogen peroxide (H(2)O(2)) was tested in an acute toxicity model with osteogenic, inflammatory, and endothelial cells. In the human fetal osteoblastic cell line 1.19 (hFOB), considerable morphological changes, cell shedding, and dysfunction of the respiratory chain were observed when cells were exposed to 3 mM H(2)O(2). Caspase 3 and poly-(ADP-ribose)-polymerase were not activated, suggesting that cell death occurred by necrosis. Preincubation of osteogenic cells, leukocytes, or endothelial cells with PRS decreased the acute toxicity of H(2)O(2). The capacity of platelets to release H(2)O(2)-detoxifying activity was retained for up to 72 h. Aminotriazole, an inhibitor of catalase, decreased the cytoprotective activity of PRS, whereas blocking of glutathione peroxidase by mercaptosuccinate had no effect. These results suggest that platelet-released catalase can rapidly neutralize cytotoxic amounts of H(2)O(2), a process that may play a role during the early stages of bone regeneration. PMID- 15784191 TI - New technology to better convey your mood (and research). PMID- 15784192 TI - Drink driving in Belgium: results from the third and improved roadside survey. AB - In 2003, the Belgian Road Safety Institute organised the third national roadside survey to estimate the proportion of drink drivers and their profile. The objective of this initiative is to gather data as a basis to formulate theory- and research-based recommendations to policymakers with the intention of decreasing the number of alcohol related accidents and victims on Belgian roads. Almost all Belgian police forces agreed to participate in a stratified two-stage cluster sample. First stage of the survey consisted of randomly selecting road sites (m = 449) in each region using a Geographical Information System (Arcview). Second stage of the survey consisted of randomly stopping drivers of personal cars (n = 12,891) during October and November 2003. All stopped drivers were asked by the police to perform an alcohol breath test. In addition, the police invited all sampled drivers to participate in a short questionnaire with individual variables (gender, age, etc.). Questionnaires with aggregated variables for road sites (traffic flow, intensity of stopping drivers, etc.) were also completed. The percentage of drivers who were found to have a blood alcohol concentration at or above the legal limit of 0.5 g/l during weekend nights (7.68%) is significantly higher than all other time spans. These percentages for the remaining time spans do not differ significantly (weekdays: 1.76%; weekday nights: 2.99%; weekend days: 2.98%). A multilevel logistic model for Belgium was successfully fitted. PMID- 15784193 TI - Visual timing and adaptive behavior in a road-crossing simulation study. AB - In this road-crossing simulation study, we assessed both participant's ability to visually judge whether or not they could cross a road, and their adaptive walking behavior. To this end, participants were presented with a road inside the laboratory on which a bike approached with different velocities from different distances. Eight children aged 5-7, ten children aged 10-12, and ten adults were asked both to verbally judge whether they could cross the road, and to actually walk across the road if possible. The results indicated that the verbal judgments were not similar to judgments to actually cross the road. With respect to safety and accuracy of judgments, groups did not differ from each other, although the youngest group tended to be more cautious. All groups appeared to use a strategy to cross the road based both on the distance and the velocity of the approaching bike. Young children waited longer on the curb before crossing the road than older children and adults. All groups adjusted their crossing time to the time-to arrival of the bike. These findings are discussed in relation to the ecological psychological approach and the putative dissociation between vision for perception (i.e. verbal judgment) and vision for action (i.e. actual crossing). PMID- 15784194 TI - Intelligent speed adaptation: accident savings and cost-benefit analysis. AB - The UK External Vehicle Speed Control (EVSC) project has made a prediction of the accident savings with intelligent speed adaptation (ISA), and estimated the costs and benefits of national implementation. The best prediction of accident reduction was that the fitting on all vehicles of a simple mandatory system, with which it would be impossible for vehicles to exceed the speed limit, would save 20% of injury accidents and 37% of fatal accidents. A more complex version of the mandatory system, including a capability to respond to current network and weather conditions, would result in a reduction of 36% in injury accidents and 59% in fatal accidents. The implementation path recommended by the project would lead to compulsory usage in 2019. The cost-benefit analysis carried out showed that the benefit-cost ratios for this implementation strategy were in a range from 7.9 to 15.4, i.e. the payback for the system could be up to 15 times the cost of implementing and running it. PMID- 15784195 TI - Exploring the overall and specific crash severity levels at signalized intersections. AB - Many studies have shown that intersections are among the most dangerous locations of a roadway network. Therefore, there is a need to understand the factors that contribute to injuries at such locations. This paper addresses the different factors that affect crash injury severity at signalized intersections. It also looks into the quality and completeness of the crash data and the effect that incomplete data has on the final results. Data from multiple sources have been cross-checked to ensure the completeness of all crashes including minor crashes that are usually unreported or not coded into crash databases. The ordered probit modeling technique has been adopted in this study to account for the fact that injury levels are naturally ordered variables. The tree-based regression methodology has also been adopted in this study to explore the factors that affect each severity level. The probit model results showed that a combination of crash-specific information and intersection characteristics result in the highest prediction rate of injury level. More specifically, having a divided minor roadway or a higher speed limit on the minor roadway decreased the level of injury while crashes involving a pedestrian/bicyclist and left turn crashes had the highest probability of a more severe crash. Several regression tree models showed a difference in the significant factors that affect the different severity types. Completing the data with minor non injury crashes improved the modeling results and depicted differences when modeling the no injury crashes. PMID- 15784196 TI - Drug use and the severity of a traffic accident. AB - Several studies have showed that driving under the influence of alcohol and/or certain illicit or medicinal drugs increases the risk of a (severe) crash. Data with respect to the question whether this also leads to a more severe accident are sparse. This study examines the relationship between the use of alcohol, illicit drugs and/or medicinal drugs and the severity of an accident within a group of drivers that were involved in a crash in The Netherlands. Blood samples of 993 drivers, collected in the period from October 1998 through September 1999, were linked to accident characteristics as available from the National Transport Research Centre. The outcome measure was the severity of the accident. An accident was considered severe when the accident had resulted in hospital admission or death. All the blood samples obtained after the accident were screened for the presence of alcohol, illicit drugs (opiates, amphetamines and amphetamine-like substances, cocaine and metabolites, methadone, cannabinoids) and medicinal drugs (benzodiazepines, barbiturates and tricyclic antidepressants). The strength of the associations between exposure to the different classes of alcohol/drugs/medicines and the severity of the accident was evaluated using logistic regression analysis and were expressed as odds ratios (OR), adjusted for age, gender, time of the day, day of the week and urban area. The most frequently detected drugs were cannabinoids, benzodiazepines and cocaine. Our results showed no clear association between the use of alcohol, illicit drug and/or medicinal drug use and the severity of the accident. Given the process of obtaining blood samples from drivers involved in accidents and the retrospective nature of the study, we cannot rule out the occurrence of selection bias. Therefore, our findings need further confirmation. PMID- 15784197 TI - The effect of seating location on the injury of properly restrained children in child safety seats. AB - Properly restrained child passengers in the National Automotive Sampling System (NASS), General Estimates System (GES), were studied for the effect of seating position on the risk of injury. The analysis focused on children seated in a child safety seat in a rear seat location. A multiple logistic regression model was used to assess whether the center rear seat is a safer seating position than either of the outboard rear seats. Standard errors for the odds ratios (ORs) of injury for several correlates of injury including seating position were obtained using a jackknife procedure. This analysis of the data showed that the center rear seat is not a safer seating position than either of the outboard rear seats in terms of odds of injury (left seat OR = 0.88, 95% CI = 0.73-1.03; right seat OR = 1.03, 95% CI = 0.85-1.20). These results do not agree with those of previous studies that suggested the center rear seat is the safest position for parents to place a child safety seat. PMID- 15784198 TI - Exposure survey of motorcyclists in New South Wales. AB - This paper reports the results of an exposure survey of 794 registered motorcycle riders, with an average of 18.1 years of riding experience, in the State of New South Wales, Australia. Respondents completed two postal surveys, separated by about 6 months, that included items relating to their crash history, riding patterns, characteristics of their motorcycle, and its odometer reading. Odometer readings indicated that respondents rode a mean of 5208 km each year, and that annual exposure was related to gender, motorcycle type, and dominant riding location, time of week, and purpose. The amount of riding reported for different purposes changed with age, with older riders more likely than younger riders to ride for recreational reasons and on weekends. The mean crash rate (based on self reported crash involvement) was 0.96 crashes/100,000 km. The crash rate declined with age, was highest in the Sydney metropolitan area, was lowest for motorcycles with large engines, and was highest for trail and dual-use motorcycles. There was a relationship between annual exposure and crash risk such that riders who rode relatively little had higher crash risks (per 100,000 km travelled) than riders who rode more often. A cluster analysis identified three groups of riders with higher-than-average risks of crash involvement. PMID- 15784199 TI - Fatal red light crashes: the role of race and ethnicity. AB - There is an increasing number of motor vehicle crashes (MVCs) in the U.S. that occur at intersections. Noncompliance with traffic signals is one of the causes for such increase. In this study, we focused on Hispanic drivers. It has been shown that failing to follow traffic laws and regulations is a major cause of MVCs in Latin America. Does this driving behavior in Latin America correspond to the Hispanic community in the U.S.? Are U.S. Hispanics more likely to die in a red light incident? Are Hispanics more likely to be red light runners than other racial/ethnic groups in the U.S.? We answered these questions by taking advantage of a 1990-1996 Fatality Analysis Reporting System (FARS) file with additional racial/ethnic information. Overall, we found no differences between African American, White, and Hispanic drivers regarding red light running. We found that drinking and driving was the most important factor for red light running. However, we found evidence that some Hispanic subgroups may be more vulnerable to red light running. One such subgroup is formed by Hispanics who have no valid driver license and no record of previous driver license suspension. This study is the first to apply national, archival data to document the relevance of race/ethnicity to the red light running problem, and we hope that it might serve as an incentive for more research on this area. PMID- 15784201 TI - Fatigue, sleep restriction and driving performance. AB - We ran a randomized cross-over design study under sleep-deprived and non-sleep deprived driving conditions to test the effects of sleep restriction on real driving performance. The study was performed in a sleep laboratory and on an open French highway. Twenty-two healthy male subjects (age = 21.5 +/- 2 years; distance driven per year = 12,225 +/- 4739 km (7641 +/- 2962 miles) [mean +/- S.D.]) drove 1000 km (625 miles) over 10 h during five 105 min sessions on an open highway. Self-rated fatigue and sleepiness before each session, number of inappropriate line crossings from video recordings and simple reaction time (RT) were measured. Total crossings increased after sleep restriction (535 crossings in the sleep-restricted condition versus 66 after non-restricted sleep (incidence rate ratio (IRR): 8.1; 95% confidence interval (95% CI): 3.2-20.5; p < 0.001)), from the first driving session. The interaction between the two factors (conditionxtime of day) was also significant (F(5, 105) = 3.229; p < 0.05). Increasing sleepiness score was associated with increasing crossings during the next driving session in the sleep-restricted (IRR: 1.9; 95% CI: 1.4-2.4) but not in the non-restricted condition (IRR: 1.0; 95% CI: 0.8-1.3). Increasing self perceived fatigue was not associated with increasing crossings in either condition (IRR: 0.95; 95% CI: 0.93-0.98 and IRR: 1.0; 95% CI: 0.98-1.02). Rested subjects drove 1000 km with four shorts breaks with only a minor performance decrease. Sleep restriction induced important performance degradation even though time awake (8h) and session driving times (105 min) were relatively short. Major inter-individual differences were observed under sleep restriction. Performance degradation was associated with sleepiness and not fatigue. Sleepiness combined with fatigue significantly affected RT. Road safety campaigns should encourage drivers to avoid driving after sleep restriction, even on relatively short trips especially if they feel sleepy. PMID- 15784200 TI - Towards a general theory of driver behaviour. AB - Taylor [Taylor, D.H., 1964. Drivers' galvanic skin response and the risk of accident. Ergonomics 7, 439-451] argued that drivers attempt to maintain a constant level of anxiety when driving which Wilde [Wilde, G.J.S., 1982. The theory of risk homeostasis: implications for safety and health. Risk Anal. 2, 209 225] interpreted to be coupled to subjective estimates of the probability of collision. This theoretical paper argues that what drivers attempt to maintain is a level of task difficulty. Naatanen and Summala [Naatanen, R., Summala, H., 1976. Road User Behaviour and Traffic Accidents. North Holland/Elsevier, Amsterdam, New York] similarly rejected the concept of statistical risk as a determinant of driver behaviour, but in so doing fell back on the learning process to generate a largely automatised selection of appropriate safety margins. However it is argued here that driver behaviour cannot be acquired and executed principally in such S-R terms. The concept of task difficulty is elaborated within the framework of the task-capability interface (TCI) model, which describes the dynamic interaction between the determinants of task demand and driver capability. It is this interaction which produces different levels of task difficulty. Implications of the model are discussed regarding variation in performance, resource allocation, hierarchical decision-making and the interdependence of demand and capability. Task difficulty homeostasis is proposed as a key sub-goal in driving and speed choice is argued to be the primary solution to the problem of keeping task difficulty within selected boundaries. The relationship between task difficulty and mental workload and calibration is clarified. Evidence is cited in support of the TCI model, which clearly distinguishes task difficulty from estimates of statistical risk. However, contrary to expectation, ratings of perceived risk depart from ratings of statistical risk but track difficulty ratings almost perfectly. It now appears that feelings of risk may inform driver decision making, as Taylor originally suggested, but not in terms of risk of collision, but rather in terms of task difficulty. Finally risk homeostasis is presented as a special case of task difficulty homeostasis. PMID- 15784202 TI - The effects of vehicle model and driver behavior on risk. AB - We study the dependence of risk on vehicle type and especially on vehicle model. Here, risk is measured by the number of driver fatalities per year per million vehicles registered. We analyze both the risk to the drivers of each vehicle model and the risk the vehicle model imposes on drivers of other vehicles with which it crashes. The "combined risk" associated with each vehicle model is simply the sum of the risk-to-drivers in all kinds of crashes and the risk-to drivers-of-other-vehicles in two-vehicle crashes. We find that most car models are as safe to their drivers as most sport utility vehicles (SUVs); the increased risk of a rollover in a SUV roughly balances the higher risk for cars that collide with SUVs and pickup trucks. We find that SUVs and to a greater extent pickup trucks, impose much greater risks than cars on drivers of other vehicles; and these risks increase with increasing pickup size. The higher aggressivity of SUVs and pickups makes their combined risk higher than that of almost all cars. Effects of light truck design on their risk are revealed by the analysis of specific models: new unibody (or "crossover") SUVs appear, in preliminary analysis, to have much lower risks than the most popular truck-based SUVs. Much has been made in the past about the high risk of low-mass cars in certain kinds of collisions. We find there are other plausible explanations for this pattern of risk, which suggests that mass may not be fundamental to safety. While not conclusive, this is potentially important because improvement in fuel economy is a major goal for designers of new vehicles. We find that accounting for the most risky drivers, young males and the elderly, does not change our general results. Similarly, we find with California data that the high risk of rural driving and the high level of rural driving by pickups does not increase the risk-to-drivers of pickups relative to that for cars. However, other more subtle differences in drivers and the driving environment by vehicle type may affect our results. PMID- 15784203 TI - Responses of side impact dummies in sled tests. AB - Sled tests were conducted at a velocity of 6.7 m/s using side impact dummies (SID, BioSID, ES-2, and WorldSIDp) and the resulting biomechanical responses were compared with responses from post mortem human subjects (PMHS). Initial impact conditions were with and without pelvic offset in combination with and without padding on the impacting wall. Impact forces, thoracic trauma index, chest compression, and viscous criteria were evaluated. The probability of injury was estimated and rates of deformation were computed for each body region. Dummy responses were not always similar in terms of trend and injury criteria when compared with PMHS tests under the same initial conditions. Response variations will be of value in improving the biofidelity characteristics of dummies for crashworthiness evaluations. PMID- 15784204 TI - Odontoid fracture in motor vehicle environments. AB - The National Automotive Sampling System (NASS) and Crash Injury Research and Engineering Network (CIREN) databases were used in an analysis of odontoid fracture in motor vehicle crashes. NASS data were evaluated for the years 1996 2002, and CIREN from 1996 to 2003. Out of 58 fractures, 38 were identified in the NASS and 20 in the CIREN databases. There were 3108 weighted cases in the NASS database. Frontal impacts (11:00 to 1:00 h) were most commonly associated with the injury in both databases. Although male and female occupants sustained the injury, females were shorter in stature, older in age, lighter in weight, and crashes were less severe (lower change in velocity) when female occupants were involved in trauma. In both databases, pure odontoid fracture and facet/lamina fracture accounted for approximately one-third of the cases, and a majority of impacts were associated with changes in velocity less than 56 km/h. Although vehicle model years ranged from 1976 to 2002, recent model years were more frequently associated with CIREN data. In the CIREN database, type II odontoid fracture was the most common, but no particular mechanism of injury dominated; such information was not available in the NASS database. To ameliorate odontoid fracture, focus should be on frontal impacts. Because different types of odontoid fracture are not included in the current Abbreviated Injury Scale, appropriate coding schemes should be developed to classify this injury. The CIREN database is unique because it provides important clinical information, i.e., fracture type, and the associated mechanism of injury. The mechanism component in any epidemiologically based injury analyses is valuable to advance improvements in vehicle crashworthiness. PMID- 15784205 TI - Road safety effects of porous asphalt: a systematic review of evaluation studies. AB - This paper presents a systematic review of studies that have evaluated the effects on road safety of porous asphalt. Porous asphalt is widely used on motorways in Europe, mainly in order to reduce traffic noise and increase road capacity. A meta-analysis was made of six studies, containing a total of eighteen estimates of the effect of porous asphalt on accident rates. No clear effect on road safety of porous asphalt was found. All summary estimates of effect indicated very small changes in accident rates and very few were statistically significant at conventional levels. Studies that have evaluated the effects of porous asphalt on nine different risk factors associated with accident occurrence were also reviewed. It was found that four of the risk factors were favourably influenced by porous asphalt, three were adversely influenced, and two were not influenced by porous asphalt. The net impact of these changes in risk factors on accident occurrence cannot be predicted. On the whole, the research that has been reported so far regarding road safety effects of porous asphalt is inconclusive. The studies are not of high quality and the findings are inconsistent. PMID- 15784206 TI - Voluntary risk taking and skill deficits in young driver accidents in the UK. AB - In absolute terms, young drivers have three to four times as many accidents per year as older drivers; and even allowing for their relative numbers in the population, their accident involvement is about 2.5 times higher than older drivers. A sample of 3437 accident reports was considered, including 1296 in detail, from midland police forces in the UK, involving drivers aged 17-25, and covering the years 1994-1996 inclusive. Four types of accident were identified as being of particular concern due to their high frequency: 'cross-flow'-turns; rear end shunts; loss of control on bends; and accidents in darkness. (The term 'cross flow' is used in relation to turns to denote an intersection accident where a driver is turning across the path of oncoming traffic, i.e., left turns in the US and continental Europe, but right turns in the UK and other countries where driving on the left side of the road is the norm.) An examination of driver risk taking behaviours as revealed in police interviews gave an insight into some of the motivational factors underlying young driver behaviour. Young driver accidents of all types are found to be frequently the result of 'risk taking' factors as opposed to 'skill deficit' factors. It had previously been thought that one of the main problems that young drivers have is in the area of specific skills needed in the driving task. However, it appears that a large percentage of their accidents are purely the result of two or three failures resulting from voluntary risk taking behaviour, rather than skill deficits per se. It is shown that specific groups of young drivers can even be considered as above average in driving skills, but simultaneously have a higher accident involvement due to their voluntary decisions to take risks. PMID- 15784207 TI - The effect of Center High Mounted Stop Lamp (CHMSL) on rear-end accidents in Israel. AB - This paper examines the effect of the use of Center High Mounted Stop Lamp (CHMSL) on rear-end accidents, as reflected in Israeli police records from calendar years 1991-2002. The basic analysis, similar to that used in previous CHMSL studies, compares the involvement in accidents of passenger cars of model years 1994-1996 that are equipped with CHMSL with passenger cars of model years 1991-1993 that are not equipped with CHMSL. The number of involvements as the struck vehicle in a rear-end accident was used as the relevant measurement and the number of involvements as the striking vehicle in a rear-end accident was used as the reference measurement. The results yielded an odds ratio of 0.93. The explanation that the CHMSL is responsible for the 7% decrease is intuitively appealing and is consistent with previous findings. However, the strength of this evidence is marginal (p = 0.07). Additional analyses evaluated the model year effect in greater detail, in order to determine whether there exists a change point between 1993 and 1994 as would be expected from a CHMSL effect, or whether the effect is spurious. Detailed analyses were performed on the ratio of struck to striking involvements as well as the rates of involvement of both types. These analyses showed that (1) the chosen reference measurement is an appropriate one, but (2) the 0.93 odds ratio is quite possibly due to other reasons unrelated to the CHMSL, thus further limiting the confidence in CHMSL effectiveness. PMID- 15784208 TI - Stated preferences for the removal of physical pain resulting from permanently disabling occupational injuries. A contingent valuation study of Taiwan. AB - Within the process of calculating the true costs of illness, physical pain is a component of intangible, or human, costs. One method of estimating the monetary value of such costs is the 'contingent valuation method' (CVM), a stated preference method based upon the elicitation of levels of willingness to pay (WTP) facilitated through surveys. This study is amongst the first of its kind to apply CVM to the estimation of the cost of the removal of physical pain resulting from permanently disabling occupational injuries. We assume that a painkilling drug has been invented to mitigate physical pain with the advantages of validity and instantaneity, and without any side effects. The WTP of each of the respondents is determined by a two-step sequential-bidding process. The maximum WTP under log normal distribution was NT 1791 US dollars/day (65.1 US dollars), whilst under Weibull distribution it was NT 1913 US dollars/day (69.6 US dollars). Older respondents, those with higher household income, fall injuries, longer periods of hospitalization, or with a perceived demand for the painkilling drug in excess of one day, displayed a positive independent effect on the eliciting of their WTP. In addition, respondents with higher 'out-of-pocket' expenses, or where the interview took place 2 years or more after the injury occurred, responded with a lower WTP. PMID- 15784209 TI - Road safety impact of extended drinking hours in Ontario. AB - PURPOSE: On 1 May 1996, Ontario, Canada amended the Liquor Licence Act to extend the hours of alcohol sales and service in licensed establishments from 1 to 2 a.m. The purpose of this study was to evaluate the road safety impact of extended drinking hours in Ontario. METHOD: A quasi-experimental design using interrupted time series with a nonequivalent no-intervention control group was used to assess changes. The analyzed data sets are total and alcohol-related, monthly, traffic fatalities for Ontario, for the 11-12 p.m., 12-1 a.m., 1-2 a.m. and 2-3 a.m. time windows, for Sunday through Wednesday nights and for Thursday through Saturday nights, for 4 years pre- and 3 years post-policy change, compared to neighbouring regions of New York and Michigan. RESULTS: The blood alcohol concentration positive driver fatality trends reflected downward trends for Sunday-Wednesday 12 2 a.m. and Thursday-Saturday 1-2 a.m. for Ontario and downward trends for Thursday-Saturday 12-1 a.m. and 2-3 a.m. for New York and Michigan after the extended drinking hour policy change. Ontario total fatality data showed similar trends to the Ontario blood alcohol positive trends. CONCLUSIONS: The multiple datasets converge in suggesting little impact on BAC positive fatalities with extension of the closing hours. These observations are consistent with other studies of small changes in alcohol availability. PMID- 15784210 TI - Parent-imposed limits on high-risk adolescent driving: are they stricter with graduated driver licensing? AB - The purpose of this study was to determine whether parent-imposed limits on 16 year-old high-risk driving are stricter in Maryland (MD), a state with graduated driver licensing (GDL) than in Connecticut (CT), a non-GDL state. In both states, parents and adolescents completed telephone surveys about the restrictions that parents placed on their adolescents' driving at night, with adolescent passengers, and at high speeds. In Maryland, surveys took place 1 month (294 parent-adolescent pairs) and 4 months (292 parent-adolescent pairs) after provisional licensure. In Connecticut, surveys took place the first month (132 pairs) and the third month (108 pairs) after adolescent licensure. The findings indicated that after controlling for demographic characteristics, Maryland parents and adolescents reported stricter parent-imposed limits for adolescent passengers, high-speed roads, weekend night driving, and overall limits. Parents in GDL states appear better able to establish and enforce adolescent driving restrictions when the licensing state stipulates, favors, and supports regulated adolescent driving. PMID- 15784211 TI - The association between sudden illness and motor vehicle crash mortality and injury among older drivers in NSW, Australia. AB - This exploratory study aims to investigate the associations between sudden illness and the risk of motor vehicle crash-related mortalities and injuries among older drivers aged 60 or above. Information utilised in the study was obtained from police reports of all road traffic accidents that occurred on the roads between 1996 and 2000 in New South Wales, Australia. There were 409 older drivers involved in crashes after becoming ill suddenly. Among these the majority (62.1%) of crashes led to at least one occupant in the vehicle being killed or injured. There is a significant association between sudden illness and crash related mortality and injury after adjusting for other risk factors. The risk of injury and death is increased by nearly six times (OR = 5.58, 95% CI = 4.54-6.85) for those who suffered a sudden illness while driving when compared to those non sufferers. These results are discussed in the light of possible preventive strategies and the provision of risk assessment and safety counselling for older people. PMID- 15784212 TI - The Cochrane Collaboration and bicycle helmets. AB - Effective interventions for care of health need to be based on scientific evidence. To this end, the Cochrane Collaboration insists that its reviews should be based on reliable data, normally obtained by randomised controlled trial. To constitute evidence, data should also support a hypothesis in accord with scientific laws and knowledge. From these considerations, an appraisal is made of the conclusion of the Cochrane review Helmets for preventing head and facial injuries in bicyclists, that it establishes scientific evidence that all types of standard helmet protect against injuries to the brain. It is concluded that the review takes no account of scientific knowledge of types and mechanisms of brain injury. It provides, at best, evidence that hard-shell helmets, now rarely used, protect the brain from injury consequent upon damage to the skull. The review therefore is not a reliable guide to the efficacy of helmets and to interventions concerning their use. PMID- 15784213 TI - Effects of a rumble strip barrier line on lane keeping in a curve. AB - This field study was conducted along a curve where the barrier line was worn out due to encroachments by cars. Vehicles were unobtrusively video recorded with the worn barrier line, after re-painting the barrier line and after milling the rumble strip barrier line. Lateral positions and speeds of vehicles were measured. Vehicle type and traffic situation were also classified (n = 6599). It was assumed that if encroachments of barrier line and the standard deviation of lateral positions were reduced, safety would improve. The standard deviation of the lateral position of passenger cars travelling in free-flow traffic decreased from 35 cm with the worn barrier line to 28 cm after re-painting the barrier line and to 24 cm after milling the rumble strip barrier line. The mean speeds of cars in free-flow traffic did not change. No vehicle with oncoming traffic crossed the barrier line after rumble strip. Encroachments of passenger cars in free-flow traffic with no oncoming traffic fell from 9.2% to 2.5% after re-painting, with no further change after implementing the rumble strip. The improvement in visual guidance alone reduced the number of crossings. However, including rumble strips may reduce the need for frequent re-painting of barriers. PMID- 15784214 TI - Child restraint system use and misuse in six states. AB - This project addressed use and misuse of child restraint systems (CRS) in the nation. CRS use and critical misuse observations were collected in the Fall of 2002 for 5527 children less than 36 kg (80 pounds) in 4126 vehicles in six states: Arizona, Florida, Mississippi, Missouri, Pennsylvania, and Washington. Results showed that 62.3% of these children were restrained in a CRS; 25.9% were restrained in a safety belt (SB); and 11.8% were unrestrained. By weight class, CRS use was 97.1% for children less than 9 kg (20 pounds); 86.4% for children between 9 and 18 kg (20 and 40 pounds); 41.7% for children between 18 and 27 kg (40 and 60 pounds); and 10.9% for children between 27 and 36 kg (60 and 80 pounds). Overall critical CRS misuse was 72.6%. Most common critical misuses were loose harness straps and loose vehicle SB attachment to the CRS. Other types of CRS misuses were also observed and recorded in the study. Recommendations are provided for field observation techniques, periodic monitoring, and research for education and enforcement strategies. PMID- 15784215 TI - [Fast tract rehabilitation in colonic surgery]. PMID- 15784216 TI - [Surgical management of GIST in the era of Gleevec]. AB - The diagnostic and treatment options for patients with GIST have evolved rapidly with the discovery of uncontrolled KIT tyrosine kinase and Gleevec that selectively inhibits Kit. Gleevec has already revolutionized the treatment of patients with metastatic disease and is also currently being tested as an adjuvant therapy after the resection of primary GIST. But the majority of responses are limited to partial responses and secondary resistances are emerging. These observations suggest that initial surgical resection remains a vital component of the treatment for patients with primary resectable cKIT+ GISTs and raises the question of secondary surgery after Gleevec. The objective of secondary surgery is to obtain a complete remission when the response to Gleevec is maximum. Surgery should be discussed between 6 and 12 months treatment when no additional improvement is observed on 2 consecutive CT scan. Three subgroups may benefit from secondary surgery: primary unresectable tumors amenable to surgery with Gleevec even in case of complete response, huge necrotic masses before expected complication, local re-progressions. Gleevec should also be discussed when a functionnal benefit can be expected by a tumor size decrease. Surgery is being evaluated in the other responding patients. The majority of responses being limited to partial responses, best indications of surgery are when complete resection may be expected (< 10%). PMID- 15784217 TI - [Fast track rehabilitation in colonic surgery: results of a prospective trial]. AB - OBJECTIVE: In elective large bowel surgery the incidence of general complications with standard perioperative care is up to 27%. Hospital discharge occurs 10 to 15 days after a conventional or laparoscopic colonic resection. The aim of a fast track management is to reduce the number of general complications and the length of hospital stay. MATERIAL AND METHODS: We prospectively evaluated a multimodal protocol in our service utilizing a combined thoracic epidural analgesia, an early mobilization and oral nutrition to accelerate postoperative recovery after elective colonic surgery. RESULTS: One hundred thirty-two consecutive patients aged an average of 66 years (range 22-88) were operated by laparotomy (n =71) or laparoscopy (n =61) and treated with the fast track rehabilitation protocol. Surgical complications occurred in 15 patients (11 %), four of these had an anastomotique leakage (3%). General complications occurred in 11 patients (8 %), the mortality was 1 %. The median length of hospital stay was four days (range 3 77) and 14 patients (11%) had to be readmitted. CONCLUSION: Application of a fast track rehabilitation protocol lowered the number of general complications and reduced the duration of hospital stay in our study. PMID- 15784218 TI - [Paragangliomas: clinical and secretory profile. Result of 39 cases]. AB - THIS RETROSPECTIVE STUDY AIMS: To define a clinical and secretory profile of paragangliomas extra-adrenal chromaffin tumors. METHODS: From 1971 throughout 2002, 39 paragangliomas have been observed in 38 patients (22 male, 16 female, average age 41,2 years). RESULTS: Four were located above the diaphragm, 35 were sub-phrenic (6 of the organ of Zuckerkandl), 32 secreted catecholamines, 23 were hypertensive (with only one without hypersecretion of catecholamines). Among 29 (131)I-metaiodobenzylguanidine scans (MIBG) reviewed, 20 tumors took up the radiopharmaceutical. The treatment was surgical in 35 cases with addition of external radiotherapy and MIBG in one case each; two patients died before any treatment. Two patients with persistent disease after surgery were successfully treated by surgery or MIBG. Histologically, 20 were malignant and 17 were seemingly benign. All exclusive dopamine secreting paragangliomas were malignant. Six patients relapsed two of which for a tumor initially classified as benign. The treatment of recurrences was surgical, by MIBG or by external radiotherapy. Nine patients had a family history of chromaffin tumor(s). The genetic survey made in five of these nine patients was positive in all cases. PMID- 15784219 TI - [CT-Guided transgluteal approach for percutaneous drainage of pelvic abscesses: results in 21 patients]. AB - OBJECTIVE: Percutaneous drainage of pelvic abscesses may be challenging using a conventional anterior route because of overlying intestinal or vascular structures. Although, the posterior transgluteal route is not commonly performed, it may provide a safer approach when the conventional anterior route is not feasible. We retrospectively analyzed our experience in transgluteal percutaneous drainage of pelvic abscesses in 21 patients to determine the feasibility, safety, tolerance and efficacy of this technique. METHODS: The data of 21 patients with pelvic abscesses (15 postoperative and 6 secondary to diverticulitis) who were treated by CT-guided percutaneous transgluteal drainage between 1992 and 2002 were reviewed. Transgluteal drainage was considered as failure in case of persisting clinical sepsis, recurrence of abscess or when surgery was needed. RESULTS: The procedure was well tolerated in all patients. No major complication was observed. In one patient mild hematoma of the piriform muscle was noticed on postprocedure CT scan but did not require a specific treatment. Successful drainage as documented by follow-up CT examination was observed in 20 patients (20/21; 95%). One case of recurrence which was successfully treated by repeated percutaneous transgluteal drainage was observed in our series. CONCLUSION: Percutaneous imaging-guided transgluteal drainage is a feasible, safe, well tolerated and effective method for the treatment of pelvic abscess when a conventional anterior route is not feasible. PMID- 15784221 TI - [How to treat common bile duct stones?]. PMID- 15784220 TI - [Is procalcitonin able to help in pediatric appendicitis diagnosis?]. AB - BACKGROUND: Appendicitis clinical diagnosis in children is difficult and the acute inflammatory markers hyperleucocytis and CRP do not contribute to it. OBJECTIVE: To study the predictive value of procalcitonin (PCT) and to assess variations of its serum level according to inflammatory lesions in paediatric appendicitis. PATIENTS AND METHOD: Among 101 children aged two and 15, hospitalized for painful abdominal syndromes, 70 underwent operation for acute appendicitis and appendiceal peritonitis, 31 had no operation. PCT was evaluated in all children, normal serum concentration was <0,5 microg/L. Every appendicectomy part has been examined through histological analysis to confirm the diagnosis and the histological lesions have been classified into three categories according to their severity. RESULTS: Out of 70 appendix analysed at histology, 68 (97%) were healthy and 2 (3%)were unhealthy. Among the 68 cases of healthy appendix, 19 had a PCT higher than 0,5 microg/L and among 33 cases of unhealthy appendix 4 had a PCT higher than 0,5 microg/L. The PCT mean of healthy children was not significantly different from unhealthy appendix population, appendicitis prevalence was 0,67, PCT sensitivity 28%: IC 95% [18-40], specificity 88%: IC 95% [72-97], positive predictive value 83%: IC 95% [61-95], and negative predictive value 37%: IC 95% [26-49]. PCT mean increases with the severity of inflammatory lesions of the appendix (P=0,0051). CONCLUSION: PCT has not a good predictive value in acute paediatric appendicitis but remains a good indicator of severity in paediatric appendicitis. PMID- 15784223 TI - [Relapse of gigantomastia after mammoplasty. Report of a case and literature review]. AB - Gigantomastia is a very rare entity of undetermined aetiology that may be due to hormonal imbalance, decreased hormonal catabolism or hypersensitivity of the target organ. It poses the problem of surgical treatment, which can be exceptionally radical of necessity. We report a case of a 32-year-old woman of whom volume and vasculocutaneous complications required a simple bilateral mastectomy to treat a recurrence of gigantomastia occurring some months after a mammoplasty. Most of the cases of gigantomastia found in the literature are associated to the pregnancy or puberty and very rare cases of spontaneous gigantomastia were listed. PMID- 15784222 TI - [Rupture of a pancreaticoduodenal artery aneurysm with median arcuate ligament syndrome: report of a case]. AB - Aneurysms of pancreaticoduodenal arteries represent only 2% of all aneurysms of digestive arteries. Occurrence of these aneurysms are favoured by stenosis or occlusion of the celiac axis. Aneurysm rupture is frequent and carries a mortality rate of 20%. Computed tomography with intravenous contrast and selective coeliomesenteric arteriography can make the diagnosis of this disease which can be treated by either surgery or embolotherapy. We report the case of a patient with a ruptured aneurysm of the pancreaticoduodenal arcades mimicking a perforated duodenal ulcer, and successfully treated by surgical ligation. PMID- 15784224 TI - [A false peritoneal carcinomatosis]. AB - Abdominal actinomycosis is an unusual peritoneal disease due to actinomyces, which are Gram-positive bacillus bacteria. Those saprophytes pathogens of digestive tract and genital mucous, can appear accidentally in the peritoneum cavity, in tumor form, simulating a neoplasic disease. We report the case of a 43 year old woman, with abdominal actinomycosis, revealed by feverish bowel obstruction. During laparotomy, several nodes were removed. Their histologic study permitted the diagnosis. Long-term antibiotics treatment allowed patient recovery. PMID- 15784225 TI - [Traumatic pancreatic injuries]. AB - If an emergency laparotomy is necessary, a damage control laparotomy may be useful. If during the laparotomy the hemodynamic is stabilised, the severity is depending on the existence of a ductal injury and an associated duodenal lesion. Surgical indications and techniques are described in these different cases. If no laparotomy, the location and type of injury is assessed by CT scan, magnetic resonance cholangiopancreatography or ERCP. Injury of the pancreatic duct is the main part of prognosis and indications. The non operative treatment in case of ductal injury remains controversial. PMID- 15784226 TI - [Simple technique for laparoscopic incisions closure]. PMID- 15784227 TI - Hydrophilic carotenoids: surface properties and aggregation behavior of a highly unsaturated carotenoid lysophospholipid. AB - The water dispersibility of a hydrophobic carotenoid has been greatly enhanced by using it as the acyl part in the synthesis of a highly unsaturated lysophospholipid. Dynamic light scattering has revealed the formation of stable aggregates with an average hydrodynamic radius of a few nanometers, and absorption spectra show that the aggregates can withstand the addition of ethanol or acetonitrile until the volume fraction of water falls below 70 and 62%, respectively. The properties of the carotenoid phospholipids have been characterized by determining surface tension, critical micelle concentration, surface concentration, molecular area, free energy of adsorption and micellation, adsorption-micellar energy relationship, and equilibrium constants. PMID- 15784228 TI - The interaction of meso-tetraphenylporphyrin with phospholipid monolayers. AB - In this article, we investigate the interaction of meso-tetraphenylporphyrin (TPP) with phospholipid monolayers. Pure TPP molecules form films at the air water interface with large extension of aggregation, which is confirmed by UV-vis spectra of transferred monolayers. For mixed films of TPP with dipalmitoyl phosphatidyl choline (DPPC) or dipalmitoyl phosphatidyl glycerol (DPPG), on the other hand, aggregation is only significant at high surface pressures or high concentrations of TPP (above 0.1 molar ratio). This was observed via Brewster angle microscopy (BAM) for the Langmuir films and UV-vis spectroscopy for transferred layers onto solid substrates. TPP indeed causes the DPPC and DPPG monolayers to expand, especially at the liquid-expanded to liquid-condensed phase transition for DPPC. The effects from TPP cannot be explained using purely geometrical considerations, as the area per TPP molecule obtained from the isotherms is at least twice the expected value from the literature. Therefore, interaction between TPP and DPPC or DPPG should be cooperative, so that more phospholipid molecules are affected than just the first neighbors to a TPP molecule. PMID- 15784229 TI - Interaction of ferulic acid derivatives with human erythrocytes monitored by pulse field gradient NMR diffusion and NMR relaxation studies. AB - Ferulic acid (Fer), a natural anti-oxidant and chemo-protector, is able to suppress experimental carcinogenesis in the forestomach, lungs, skin, tongue and colon. Several Fer derivatives have been suggested as promising candidates for cancer prevention, being the biological activity related also to the capacity of partitioning between aqueous and lipid phases. In the present work, pulsed field gradient (PFG) NMR diffusion measurement and NMR relaxation rates have been adopted for investigating the interaction of three Fer derivatives (Fer-C11, Fer C12 and Fer-C13) with human erythrocytes. Binding to the erythrocyte membrane has been shown for all derivatives, which displayed a similar interaction mode such that the aromatic moiety and the terminal part of the alkyl chain were the most affected. Quantitative analysis of the diffusion coefficients was used to show that Fer-C12 and Fer-C13 display higher affinity for the cell membrane when compared with Fer-C11. These findings agree with the higher anti-oxidant activity of the two derivatives. PMID- 15784230 TI - Raman spectroscopy investigation of various saturated monoacid triglycerides. AB - A study of the vibrational behavior of five saturated monoacid triacylglycerides is performed by Raman spectroscopy at room temperature in the 3100-500 cm(-1) spectral range. The splitting of the CO stretching mode leads to conclude on the existence of two or three geometries of CO in the "knot" group OCO. The CO stretching mode seems to be a good tool for distinguishing the polymorphic forms of the studied triglycerides. The assignments of the different CH stretching modes are performed in the 1500-500 cm(-1) spectral range. The I(2845)/I(2880) (in the CH stretching spectral region) and I(1445)/I(1296) intensity ratios (between the maximal intensity I(1445) of the CH(2) scissoring mode and the maximal intensity I(1296) of the skeletal vibration of (CH(2))(n) in-phase twist) seem to depend on the type of polymorphic forms of these molecules. PMID- 15784231 TI - Measurement of saturable and non-saturable components of anandamide uptake into P19 embryonic carcinoma cells in the presence of fatty acid-free bovine serum albumin. AB - There is considerable controversy at present concerning the mechanisms responsible for the cellular uptake of anandamide. One particular issue concerns whether fatty acid-free bovine serum albumin should be used in the assays, it having been argued that such a presence effectively prevents the specific uptake of anandamide. In the present study, it has been demonstrated that in the presence of a low (0.1%, w/v) concentration of fatty acid-free bovine serum albumin, a temperature-dependent and saturable (K(m) approximately 1 microM) uptake of anandamide into P19 embryonic carcinoma cells can be demonstrated using an incubation time of 4 min. Under these conditions, the uptake of anandamide at 4 degrees C is low at a substrate concentration of 100 nM. The uptake at 37 degrees C was not significantly reduced following treatment of the cells with either methyl-beta-cyclodextrin (50 microM) or mevinolin (1 microM), but was reduced by the FAAH inhibitor URB597 (1 microM) and inhibited by the transport inhibitor cum FAAH substrate AM404 with an IC(50) value of 12 microM. When a 45 s incubation time was used, the uptake of anandamide was not saturable at 37 degrees C over the concentration range tested (0.1-1 microM). Analysis of the data at 37 degrees C obtained with 45 s, 4 min and 15 min incubation times revealed a very rapid (i.e. complete by 45 s) non-saturable component followed by a slower saturable (K(m) approximately 1 microM) component of the uptake. It is concluded that the presence of a low concentration of fatty acid-free bovine serum albumin at a suitable concentration reduces non-specific binding (and release) of anandamide to cell culture wells, greatly reduces the cellular accumulation seen at 4 degrees C, and allows the visualisation of both non saturable and saturable components of the uptake to be seen at 37 degrees C. PMID- 15784232 TI - Chemical synthesis of 24-beta-D-galactopyranosides of bile acids: a new type of bile acid conjugates in human urine. AB - A method is reported for the preparation of the C-24 carboxyl-linked beta-D galactopyranosides of lithocholic, deoxycholic, chenodeoxycholic, ursodeoxycholic, and cholic acids, two of which were recently identified as a novel type of the metabolites of bile acids excreted in human urine. Direct esterification (galactosidation) of the unprotected bile acids with 2,3,4,6-tetra O-benzyl-D-galactopyranose in the presence of 2-chloro-1,3,5-trinitrobenzene as a coupling agent and subsequent hydrogenolysis of the resulting benzyloxy-protected bile acid 24-beta-D-galactopyranosides over 10% palladium on charcoal under atmospheric pressure afforded the title compounds. The structures of the bile acid acyl galactosides were confirmed by measuring several (1)H-(1)H and (1)H (13)C shift correlated 2D NMR. PMID- 15784233 TI - Headgroup effects on phase behavior and interfacial properties of beta-3,7 dimethyloctylglycoside/water systems. AB - The aqueous phase behavior and the interfacial properties of alkylglycosides, AGs, with a 3,7-dimethyloctyl (geranyl) chain have been investigated as a function of the number of glucose units, N, in the maltooligosaccharide headgroup. The results can be interpreted in terms of a "helical conformation" of the maltooligosaccharide, where the cross section area increases as N increases. PMID- 15784234 TI - Free radical oxidation of coriolic acid (13-(S)-hydroxy-9Z,11E-octadecadienoic acid). AB - The reaction of (13S,9Z,11E)-13-hydroxy-9,11-octadecadienoic acid (1a), one of the major peroxidation products of linoleic acid and an important physiological mediator, with the Fenton reagent (Fe(2+)/EDTA/H(2)O(2)) was investigated. In phosphate buffer, pH 7.4, the reaction proceeded with >80% substrate consumption after 4h to give a defined pattern of products, the major of which were isolated as methyl esters and were subjected to complete spectral characterization. The less polar product was identified as (9Z,11E)-13-oxo-9,11-octadecadienoate (2) methyl ester (40% yield). Based on 2D NMR analysis the other two major products were formulated as (11E)-9,10-epoxy-13-hydroxy-11-octadecenoate (3) methyl ester (15% yield) and (10E)-9-hydroxy-13-oxo-10-octadecenoate (4) methyl ester (10% yield). Mechanistic experiments, including deuterium labeling, were consistent with a free radical oxidation pathway involving as the primary event H-atom abstraction at C-13, as inferred from loss of the original S configuration in the reaction products. Overall, these results provide the first insight into the products formed by oxidation of 1a with the Fenton reagent, and hint at novel formation pathways of the hydroxyepoxide 3 and hydroxyketone 4 of potential (patho)physiological relevance in settings of oxidative stress. PMID- 15784235 TI - Direct monitoring of lipid oxidation in edible oils by Fourier transform Raman spectroscopy. AB - Fourier transform Raman spectroscopy has been used to investigate the chemical changes taking place during lipid oxidation in several edible oils. Oxidative degradation of six vegetable oils was accelerated by heating at 160 degrees C. Formation of aldehydes was detected, and saturated as well as alpha,beta unsaturated aldehydes could be identified with the help of pure component spectra. The formation of conjugated double bond systems and the isomerisation of cis to trans double bonds was observed in the C=C stretching region and found to follow a distinct pattern for the different oils. It was possible to associate these differences to the fatty acid composition. The time-dependent intensity changes in certain Raman bands were compared to conventional parameters used to determine the extent of oxidation in oils, such as anisidine value and K(270), and showed good correlation. PMID- 15784236 TI - Cholesterol inhibits the lytic activity of melittin in erythrocytes. AB - Although cell lysis by the hemolytic peptide, melittin, has been extensively studied, the role of specific lipids of the erythrocyte membrane on melittin induced hemolysis remains unexplored. In this report, we have explored the modulatory role of cholesterol on the hemolytic activity of melittin by specifically depleting cholesterol from rat erythrocytes using methyl-beta cyclodextrin (MbetaCD). Our results show that the hemolytic activity of melittin is increased by approximately 3-fold upon depletion of erythrocyte membrane cholesterol by approximately 55% without any appreciable loss of phospholipids. This result constitutes the first report demonstrating that the presence of cholesterol inhibits the lytic activity of melittin in its natural target membrane, i.e., the erythrocyte membrane. These results are relevant in understanding the role of cholesterol in the mechanism of action of melittin in the erythrocyte membrane. PMID- 15784238 TI - Synthesis and anti-inflammatory activity of 1-acylaminoalkyl-3,4-dialkoxybenzene derivatives. AB - New 1-acylaminoalkyl-3,4-dialkoxybenzene derivatives 17-31 were synthesized by the acylation of amines 9-16 with acyl chlorides. Amines 9-16 were obtained from aryl ketones 1-8. Aryl ketones 1-8 were synthesized by the acylation of corresponding aromatic compounds. As it was preliminary predicted by PASS (Prediction of Activity Spectra for Substance) program, all 1-acylaminoalkyl-3,4 dimethoxy- and 3,4-diethoxybenzene derivatives possess anti-inflammatory activity. Activity of compounds 18, 19, 21, 24, 26, 27, 28, 29 was similar to that of acetylsalicylic acid or ibuprofen however their acute toxicity was less than that of mentioned anti-inflammatory drugs. A series of 1-acylaminoalkyl-3,4 dimethoxybenzene, 1-acylaminoalkyl-3,4-diethoxybenzene and 6-acylaminoalkyl-2,3 dihydro-1,4-benzodioxine derivatives have been synthesized. These compounds possess moderate or strong anti-inflammatory activity and low toxicity. PMID- 15784237 TI - New heterocyclic ligands for the adenosine receptors P1 and for the ATP receptors P2. AB - Extracellular adenosine and adenine nucleotides induce various cellular responses through activation of P1 and P2 receptors. P1 receptors preferentially recognize adenosine and four different G protein-coupled receptors (A(1), A(2A), A(2B), and A(3) subtypes) have been identified. On the other hand, P2 receptors are activated by adenine and/or uridine nucleotides and classified into two families: ionotropic P2X and G protein-coupled P2Y receptors. In this article, we summarize our studies which led to development of new potent and selective heterocyclic ligands for the adenosine receptors P1 and for the ATP receptors P2X(7). PMID- 15784239 TI - 3D-QSAR and preliminary evaluation of anti-inflammatory activity of series of N pyrrolylcarboxylic acids. AB - The present study focuses on development of new potential inhibitors of cyclooxygenase-2 (COX-2): series of N-pyrrolylcarboxylic acids. 3D-QSAR (Quantitative Structure-Activity Relationship) CoMFA (Comparative Molecular Field Analysis) and CoMSIA (Comparative Molecular Similarity Index Analysis) models for predicting inhibitory activities against COX-1 and COX-2 as well as for evaluating in vivo anti-inflammatory activity were obtained and used for preliminary screening of new anti-inflammatory N-pyrrolylcarboxylic acids. Nine compounds were selected for in vivo testing and evaluated for their potency to decrease carrageenin-induced edema in rats. The compounds were applied i.p. at doses 20 mg/kg and 40 mg/kg and p.o. at doses 10 mg/kg and 40 mg/kg. Six compounds showed more than 70% protection of the edema. Indomethacin (2 mg/kg i.p.), used as a reference drug, possessed 54% anti-inflammatory activity under similar experimental conditions. PMID- 15784240 TI - Nematocidal and trichomonacidal activities of 2-substituted quinolines. AB - Several quinolines were synthesized and evaluated in vitro and in vivo against the nematodes Caenorhabditis elegans, Heligmosomoides polygyrus and the protozoa Trichomonas vaginalis. If some of them have shown in vitro nematocide activity (at 10 microM), however, their trichomonacidal activity reached 50% reduction at only 100 microM. The in vivo activity on Trichinella spiralis model was evaluated for some of the most in vitro active quinolines. PMID- 15784241 TI - Validation of the anti-factor IIa assay and potency assessment of enoxaparin in pharmaceutical formulations. AB - The low molecular weight heparin enoxaparin sodium is used clinically for the prevention and treatment of venous and arterial thrombosis. An anti-factor IIa assay was applied and validated for the potency evaluation of the pharmaceutical formulations. Investigating the parameters of range, linearity (r(2)=0.9912), precision, accuracy and robustness, the biological assay incorporated a chromogenic end-point and detection at 405 nm. The method yielded good results with a detection limit of 0.01 IU/ml and a quantitation limit of 0.03 IU/ml. Sodium enoxaparin pharmaceutical products were evaluated by the anti-factor Xa assay and the anti-factor IIa assay giving potencies between 97.18% and 106.50%, with an anti-factor Xa/anti-factor IIa ratio between 3.9 and 4.2. The results demonstrated the validity of the anti-factor IIa assay that together with the anti-factor Xa are useful methodologies for the routine quality control of enoxaparin in pharmaceutical products. PMID- 15784242 TI - Synthesis of 2-semicarbazonomethyl-4,5-methylenedioxyphenylacetic acids as anticonvulsant agents. AB - A series of 2-semicarbazonomethyl-4,5-methylenedioxyphenylacetic acids (12-19) were synthesized and tested as anticonvulsant agents in DBA/2 mice against sound induced seizures and the results compared to those previously reported for the corresponding methyl esters (4-11). The new compounds possess anticonvulsant properties lower than those of 4-11, but, in some instances, comparable to that of GYKI 52466, a well-known noncompetitive AMPA receptor antagonist. PMID- 15784243 TI - Synthesis of novel N-substituted imidazolecarboxylic acid hydrazides as monoamine oxidase inhibitors. AB - Novel 2-alkylsulfanyl-1-benzyl-5-imidazolecarboxylic acid hydrazides (15a,b) were synthesized as analogues of isocarboxazide, which is a known nonselective irreversible monoamine oxidase inhibitor and tested for monoamine oxidase A and B inhibitory activity. Neither of the compounds showed any inhibition of MAO B activity up to a high concentration of 100 microM. An MAO A activity was only slowly inhibited at this high concentration after prolonged incubation with either compound. This suggests any observed inhibition is not very specific. PMID- 15784244 TI - Synthesis of 2-N,N-dimethylaminomethyl-2,3,3a,12b-tetrahydrodibenzo[b,f]furo[2,3 d]oxepine derivatives as potential anxiolytic agents. Part 2: substitutions by methyl groups on the tetrahydrofuran ring. AB - New synthesis approaches that have led to a series of novel tetrahydrodibenzo[b,f]furo[2,3-d]oxepine derivatives are described. A systematic synthetic approach for the introduction of small carbon substituents (methyl groups) around the tetrahydrofuran moiety of tetrahydrodibenzo[b,f]furo[2,3 d]oxepine derivatives is reported. Preliminary pharmacological data of the newly synthesised compounds are also communicated. PMID- 15784245 TI - Simultaneous spectrophotometric determination of salbutamol and bromhexine in tablets. AB - Typical anti-mucolytic drugs called salbutamol hydrochloride and bromhexine sulfate encountered in tablets were determined simultaneously either by using linear regression at zero-crossing wavelengths of the first derivation of UV spectra or by application of multiple linear partial least squares regression method. The results obtained by the two proposed mathematical methods were compared with those obtained by the HPLC technique. PMID- 15784246 TI - Modeling the electrophoretic mobility of beta-blockers in capillary electrophoresis using artificial neural networks. AB - Artificial neural networks were used for modeling the mobility of five beta blockers (i.e., labetalol atenolol, practolol, timolol and propranolol) in running buffer with ternary solvent background electrolyte systems containing 80 mM acetate buffer dissolved in water, methanol, ethanol and their ternary mixtures. The volume fractions of two solvents (f(2), f(3)) and cologarithm of electrophoretic mobilities in pure solvents (i.e., -Lnmu(1), -Lnmu(2) and Lnmu(3)) were used as inputs and cologarithm of the mobility in mixed solvents was the output of the networks. The number of neurons in hidden layer, learning rate, momentum and the number of epochs were optimized, in which two neurons in hidden layer, 0.2, 0.9 and 20000 were found the optimized values for learning rate, momentum and number of epochs, respectively. Mean percentage deviations (MPD) between calculated and experimental mobilities were computed as an accuracy criterion. To assess the correlative ability of the model, all data points in each set were used as training set and the mobilities were back-calculated by the trained networks, in which the overall MPD (OMPD)+/- standard deviation (SD) for correlative study was 3.1+/- 2.3. To evaluate the prediction capability of the proposed ANN model, the network was trained using 15 data points for each analyte and the remaining data points were predicted. The obtained OMPD (+/-SD) for this analysis was 3.6+/-3.0. To further investigate on the applicability of ANN, a generalized network was trained with 10 data points from each beta-blocker and then the network was employed to predict the mobilities of the analytes in ternary solvent electrolyte systems. The MPDs for predicted mobilities were 3.6%, 3.6%, 3.9%, 3.7% and 2.9% respectively for labetalol, atenolol, practolol, timolol and propranolol. PMID- 15784247 TI - Effect of viscosity and concentration of wall former, emulsifier and pore-inducer on the properties of amoxicillin microcapsules prepared by emulsion solvent evaporation. AB - This study reports the laboratory optimization for the preparation of sustained release amoxicillin (AMX) ethylcellulose microcapsules by an emulsion solvent evaporation process by adjusting the viscosity and concentration of ethylcellulose, ratio of amoxicillin to ethylcellulose, and concentration of emulsifier and pore inducer. When ethylcellulose with a viscosity of 45 mPa.s was used, almost no material stuck to the inside wall of the beaker and uniform microcapsules were prepared. The average diameter of microcapsules increased and yield and release rate decreased as the concentration of ethylcellulose increased from 1% to 8%. The release of amoxicillin from microcapsules was influenced by the ratio of the weight of drug to that of ethylcellulose and ratios of 2:1 and 4:1 were most suited for optimum amoxicillin release. The average diameter of microcapsules decreased and the release rate increased as the concentration of the emulsifier increased from 1.5% to 6.0%, however, the size distribution became significantly wider with the increase in the concentration of sorbitan monooleate. Addition of small amounts of a water-soluble agent sucrose improved the release of active ingredient from the microcapsule matrix without influencing the morphology and particulate properties of the microcapsules. PMID- 15784248 TI - HPLC and chemometric-assisted spectrophotometric methods for simultaneous determination of atenolol, amiloride hydrochloride and chlorthalidone. AB - Three methods are presented for the simultaneous determination of atenolol (AT), amiloride hydrochloride (AM) and chlorthalidone (CD). The high performance liquid chromatographic (HPLC) method depends on the separation of each drug on a reversed phase, RP (18) column. Elution was carried out with a mobile phase consisting of acetonitrile -5mM heptansulphonic acid sodium salt (20:80, v/v, pH 4.4). Quantitation was achieved with UV detection at 274 nm based on peak area. The other two-chemometric-assisted spectrophotometric methods applied were principal component regression (PCR) and partial least squares (PLS-1). These approaches were successfully applied to quantify each drug in the mixture using the information included in the absorption spectra of appropriate solutions in the range 240-290 nm with the intervals Deltalambda=0.2 nm. The three methods were successfully applied to a pharmaceutical formulation (tablets), and the results were compared with each other. PMID- 15784249 TI - E93K charge reversal on actin perturbs steric regulation of thin filaments. AB - Contraction in striated muscles is regulated by Ca2+-dependent movement of tropomyosin-troponin on thin filaments. Interactions of charged amino acid residues between the surfaces of tropomyosin and actin are believed to play an integral role in this steric mechanism by influencing the position of tropomyosin on the filaments. To investigate this possibility further, thin filaments were isolated from troponin-regulated, indirect flight muscles of Drosophila mutants that express actin with an amino acid charge reversal at residue 93 located at the interface between actin subdomains 1 and 2, in which a lysine residue is substituted for a glutamic acid. Electron microscopy and 3D helical reconstruction were employed to evaluate the structural effects of the mutation. In the absence of Ca2+, tropomyosin was in a position that blocked the myosin binding sites on actin, as previously found with wild-type filaments. However, in the presence of Ca2+, tropomyosin position in the mutant filaments was much more variable than in the wild-type ones. In most cases (approximately 60%), tropomyosin remained in the blocking position despite the presence of Ca2+, failing to undergo a normal Ca2+-induced change in position. Thus, switching of a negative to a positive charge at position 93 on actin may stabilize negatively charged tropomyosin in the Ca2+-free state regardless of Ca2+ levels, an alteration that, in turn, is likely to interfere with steric regulation and consequently muscle activation. These results highlight the importance of actin's surface charges in determining the distribution of tropomyosin positions on thin filaments derived from troponin-regulated striated muscles. PMID- 15784250 TI - Structure of the connector of bacteriophage T7 at 8A resolution: structural homologies of a basic component of a DNA translocating machinery. AB - The three-dimensional structure of the bacteriophage T7 head-to-tail connector has been obtained at 8A resolution using cryo-electron microscopy and single particle analysis from purified recombinant connectors. The general morphology of the T7 connector is that of a 12-folded toroidal homopolymer with a channel that runs along the longitudinal axis of the particle. The structure of the T7 connector reveals many structural similarities with the connectors from other bacteriophages. Docking of the atomic structure of the varphi29 connector into the three-dimensional reconstruction of T7 connector reveals that the narrow, distal region of the two oligomers are almost identical. This region of the varphi29 connector has been suggested to be involved in DNA translocation, and is composed of an alpha-beta-alpha-beta-beta-alpha motif. A search for alpha-helices in the same region of the T7 three-dimensional map has located three alpha helices in approximately the same position as those of the varphi29 connector. A comparison of the predicted secondary structure of several bacteriophage connectors, including among others T7, varphi29, P22 and SPP1, reveals that, despite the lack of sequence homology, they seem to contain the same alpha-beta alpha-beta-beta-alpha motif as that present in the varphi29 connector. These results allow us to suggest a common architecture related to a basic component of the DNA translocating machinery for several viruses. PMID- 15784251 TI - High hydrostatic pressure dissociates early aggregates of TTR105-115, but not the mature amyloid fibrils. AB - A range of disorders such as Alzheimer's disease and type II diabetes have been linked to protein misfolding and aggregation. Transthyretin is an amyloidogenic protein which is involved in familial amyloid polyneuropathy, the most common form of systemic amyloid disease. A peptide fragment of this protein, TTR105-115, has been shown to form well-defined amyloid fibrils in vitro. In this study, the stability of amyloid fibrils towards high hydrostatic pressure has been investigated by Fourier transform infrared spectroscopy. Information on the morphology of the species exposed to high hydrostatic pressure was obtained by atomic force microscopy. The species formed early in the aggregation process were found to be dissociated by relatively low hydrostatic pressure (220 MPa), whereas mature fibrils are pressure insensitive up to 1.3 GPa. The pressure stability of the mature fibrils is consistent with a fibril structure in which there is an extensive hydrogen bond network in a tightly packed environment from which water is excluded. The fact that early aggregates can be dissociated by low pressure suggests, however, that hydrophobic and electrostatic interactions are the dominant factors stabilizing the species formed in the early stages of fibril formation. PMID- 15784252 TI - An obligatory intermediate controls the folding of the alpha-subunit of tryptophan synthase, a TIM barrel protein. AB - The proposed kinetic folding mechanism of the alpha-subunit of tryptophan synthase (alphaTS), a TIM barrel protein, displays multiple unfolded and intermediate forms which fold through four parallel pathways to reach the native state. To obtain insight into the secondary structure that stabilizes a set of late, highly populated kinetic intermediates, the refolding of urea-denatured alphaTS from Escherichia coli was monitored by pulse-quench hydrogen exchange mass spectrometry. Following dilution from 8 M urea, the protein was pulse labeled with deuterium, quenched with acid and mass analyzed by electrospray ionization mass spectrometry (ESI-MS). Hydrogen bonds that form prior to the pulse of deuterium offer protection against exchange and, therefore, retain protons at the relevant amide bonds. Consistent with the proposed refolding model, an intermediate builds up rapidly and decays slowly over the first 100 seconds of folding. ESI-MS analysis of the peptic fragments derived from alphaTS mass-labeled and quenched after two seconds of refolding indicates that the pattern of protection of the backbone amide hydrogens in this transient intermediate is very similar to that observed previously for the equilibrium intermediate of alphaTS highly populated at 3 M urea. The protection observed in a contiguous set of beta-strands and alpha-helices in the N terminus implies a significant role for this sub-domain in directing the folding of this TIM barrel protein. PMID- 15784253 TI - The nuclear receptor coactivator PGC-1alpha exhibits modes of interaction with the estrogen receptor distinct from those of SRC-1. AB - Estrogen receptor (ER) function is mediated by multi-domain co-regulator proteins. A fluorescently labelled fragment of the human PGC-1alpha co-regulator (residues 91-408) bearing the two motifs most strongly implicated in interactions with nuclear receptors (NR box2 and NR box3), was used to characterize in vitro binding of PGC-1alpha to ER. Anisotropy measurements revealed that the affinity of this PGC-1alpha fragment for human ERalpha and beta was fairly strong in the presence of estradiol (approximately 5 nM), and that unlike a similar fragment of SRC-1 (570-780), PGC-191-408 exhibited ligand-independent interactions with ER, particularly with ERbeta (Kd approximately 30 nM). Competition experiments of the complex between ERalpha and fluorescently labelled PGC-1 91-408 with unlabelled SRC-1 570-780 showed that PGC-1 91-408 was an efficient competitor of SRC-1 570 780, while the inverse was not true, underscoring their distinct modes of binding. The anisotropy data provide strong evidence for a ternary complex between ERalpha, SRC-1 570-780 and PGC-1 91-408. GST-pull-down experiments with deletion mutants of ERalpha revealed that the constitutive binding of PGC-1 91 408 requires the presence of the linker domain between the DNA binding and ligand binding domains (DBD and LBD). Homology modeling studies of the different regions of full length PGC-1alpha confirmed the lack of compact tertiary structure of the N-terminal region bearing the NR box motifs, and suggested a slightly different mode of interaction compared to the NR box motifs of SRC-1. They also provided reasonable structural models for the coiled-coil dimerization motif at residues 633-675, as well as the C-terminal putative RNA binding domain, raising important questions concerning the stoichiometry of its complex with the nuclear receptors. PMID- 15784254 TI - Domain study of bacteriophage p22 coat protein and characterization of the capsid lattice transformation by hydrogen/deuterium exchange. AB - Viral capsids are dynamic structures which undergo a series of structural transformations to form infectious viruses. The dsDNA bacteriophage P22 is used as a model system to study the assembly and maturation of icosahedral dsDNA viruses. The P22 procapsid, which is the viral capsid precursor, is assembled from coat protein with the aid of scaffolding protein. Upon DNA packaging, the capsid lattice expands and becomes a stable virion. Limited proteolysis and biochemical experiments indicated that the coat protein consists of two domains connected by a flexible loop. To investigate the properties and roles of the sub domains, we have cloned them and initiated structure and function studies. The N terminal domain, which is made up of 190 amino acid residues, is largely unstructured in solution, while the C-terminal domain, which consists of 239 amino acid residues, forms a stable non-covalent dimer. The N-terminal domain adopts additional structure in the context of the C-terminal domain which might form a platform on which the N-terminal domain can fold. The local dynamics of the coat protein in both procapsids and mature capsids was monitored by hydrogen/deuterium exchange combined with mass spectrometry. The exchange rate for C-terminal domain peptides was similar in both forms. However, the N-terminal domain was more flexible in the empty procapsid shells than in the mature capsids. The flexibility of the N-terminal domain observed in the solution persisted into the procapsid form, but was lost upon maturation. The loop region connecting the two domains exchanged rapidly in the empty procapsid shells, but more slowly in the mature capsids. The global stabilization of the N-terminal domain and the flexibility encoded in the loop region may be a key component of the maturation process. PMID- 15784255 TI - Crystal structure of maltooligosyltrehalose trehalohydrolase from Deinococcus radiodurans in complex with disaccharides. AB - Trehalose (alpha-D-glucopyranosyl-1,1-alpha-D-glucopyranose) is a non-reducing diglucoside found in various organisms that serves as a carbohydrate reserve and as an agent that protects against a variety of physical and chemical stresses. Deinococcus radiodurans possesses an alternative biosynthesis pathway for the synthesis of trehalose from maltooligosaccharides. This reaction is mediated by two enzymes: maltooligosyltrehalose synthase (MTSase) and maltooligosyltrehalose trehalohydrolase (MTHase). Here, we present the 1.1A resolution crystal structure of MTHase. It consists of three major domains: two beta-sheet domains and a conserved glycosidase (beta/alpha)8 barrel catalytic domain. Three subdomains consisting of short insertions were identified within the catalytic domain. Subsequently, structures of MTHase in complex with maltose and trehalose were obtained at 1.2 A and 1.5 A resolution, respectively. These structures reveal the importance of the three inserted subdomains in providing the key residues required for substrate recognition. Trehalose is recognised specifically in the +1 and +2 binding subsites by an extensive hydrogen-bonding network and a strong hydrophobic stacking interaction in between two aromatic residues. Moreover, upon binding to maltose, which mimics the substrate sugar chain, a major concerted conformational change traps the sugar chain in the active site. The presence of magnesium in the active site of the MTHase-maltose complex suggests that MTHase activity may be regulated by divalent cations. PMID- 15784256 TI - Evidence for structural plasticity of heavy chain complementarity-determining region 3 in antibody-ssDNA recognition. AB - Anti-DNA antibodies play important roles in the pathogenesis of autoimmune diseases. They also represent a unique and relatively unexplored class of DNA binding protein. Here, we present a study of conformational changes induced by DNA binding to an anti-ssDNA Fab known as DNA-1. Three crystal structures are reported: a complex of DNA-1 bound to dT3, and two structures of the ligand-free Fab. One of the ligand-free structures was determined from crystals exhibiting perfect hemihedral twinning, and the details of structure determination are provided. Unexpectedly, five residues (H97-H100A) in the apex of heavy chain complementarity-determining region 3 (HCDR3) are disordered in both ligand-free structures. Ligand binding also caused a 2-4A shift of the backbone of Tyr L92 and ordering of the L92 side-chain. In contrast, these residues are highly ordered in the Fab/dT3 complex, where Tyr H100 and Tyr H100A form intimate stacking interactions with DNA bases, and L92 forms the 5' end of the binding site. The structures suggest that HCDR3 is very flexible and adopts multiple conformations in the ligand-free state. These results are discussed in terms of induced fit and pre-existing equilibrium theories of ligand binding. Our results allow new interpretations of existing thermodynamic and mutagenesis data in terms of conformational entropy and the volume of conformational space accessible to HCDR3 in the ligand-free state. In the context of autoimmune disease, plasticity of the ligand-free antibody could provide a mechanism by which anti-DNA antibodies bind diverse host ligands, and thereby contribute to pathogenicity. PMID- 15784257 TI - The structure of DC-SIGNR with a portion of its repeat domain lends insights to modeling of the receptor tetramer. AB - The dendritic cell-specific ICAM-3 non-integrin (DC-SIGN) and its close relative DC-SIGNR recognize various glycoproteins, both pathogenic and cellular, through the receptor lectin domain-mediated carbohydrate recognition. While the carbohydrate-recognition domains (CRD) exist as monomers and bind individual carbohydrates with low affinity and are permissive in nature, the full-length receptors form tetramers through their repeat domain and recognize specific ligands with high affinity. To understand the tetramer-based ligand binding avidity, we determined the crystal structure of DC-SIGNR with its last repeat region. Compared to the carbohydrate-bound CRD structure, the structure revealed conformational changes in the calcium and carbohydrate coordination loops of CRD, an additional disulfide bond between the N and the C termini of the CRD, and a helical conformation for the last repeat. On the basis of the current crystal structure and other published structures with sequence homology to the repeat domain, we generated a tetramer model for DC-SIGN/R using homology modeling and propose a ligand-recognition index to identify potential receptor ligands. PMID- 15784258 TI - HCV NS3 serine protease-neutralizing single-chain antibodies isolated by a novel genetic screen. AB - Hepatitis C virus (HCV) infection is a major world-wide health problem causing chronic hepatitis, liver cirrhosis and primary liver cancer. The high frequency of treatment failure points to the need for more specific, less toxic and more active antiviral therapies for HCV. The HCV NS3 is currently regarded as a prime target for anti-viral drugs, thus specific inhibitors of its activity are of utmost importance. Here, we report the development of a novel bacterial genetic screen for inhibitors of NS3 catalysis and its application for the isolation of single-chain antibody-inhibitors. Our screen is based on the concerted co expression of a reporter gene, of recombinant NS3 protease and of fusion stabilized single-chain antibodies (scFvs) in Escherichia coli. The reporter system had been constructed by inserting a short peptide corresponding to the NS5A/B cleavage site of NS3 into a permissive site of the enzyme beta galactosidase. The resulting engineered lacZ gene, coding for an NS3-cleavable beta-galactosidase, is carried on a low copy plasmid that also carried the NS3 protease-coding sequence. The resultant beta-galactosidase enzyme is active, conferring a Lac+ phenotype (blue colonies on indicator 5-bromo-4-chloro-3 indolyl beta-D-galactoside (X-gal) plates), while induction of NS3 expression results in loss of beta-galactosidase activity (transparent colonies on X-gal plates). The identification of inhibitors, as shown here by isolating NS3 inhibiting single-chain antibodies, expressed from a compatible high copy number plasmid, is based on the appearance of blue colonies (NS3 inhibited) on the background of colorless colonies (NS3 active). Our source of inhibitory scFvs was an scFv library that we prepared from spleens of NS3-immunized mice and subjected to limited affinity selection. Once isolated, the inhibitors were validated as genuine and specific NS3 binders by an enzyme-linked immunosorbent assay and as bone fide NS3 serine protease inhibitors by an in vitro catalysis assay. We further show that upon expression as cytoplasmic intracellular antibodies (intrabodies) in NS3-expressing mammalian cells, three of the scFvs inhibit NS3 mediated cell proliferation. Although applied here for the isolation of antibody based inhibitors, our genetic screen should be applicable for the identification of candidate inhibitors from other sources. PMID- 15784260 TI - Molecular crowding limits the role of fetal hemoglobin in therapy for sickle cell disease. AB - The dominant assumption central to most treatments for sickle cell anemia has been that replacement of sickle hemoglobin (HbS) by fetal hemoglobin (HbF) would have major clinical benefit. Using laser photolysis, we have measured polymerization kinetics including rates of homogeneous and heterogeneous nucleation on mixtures of 20% and 30% HbF with HbS. We find that the present model for polymerization, including molecular crowding, can accurately predict the rates of such mixtures, by using the single assumption that no significant amount of HbF enters the polymer. The effects of replacing HbS by HbF on the rates of polymer formation are found to be significantly lower than previous measurements appeared to indicate because the impact of the replacement is also highly dependent on the total hemoglobin concentration. This is because the molecular crowding of non-polymerizing HbF offsets substantially the effects of decreasing the concentration of HbS concentration, an effect that increases with concentration. Most strikingly, the demonstrated benefit of hydroxyurea therapy in slowing the kinetics of intracellular polymerization cannot be primarily due to enhanced HbF, but must have some other origin, which could itself represent a promising therapeutic approach. PMID- 15784259 TI - The 1.1 A resolution crystal structure of the p130cas SH3 domain and ramifications for ligand selectivity. AB - The Crk-associated tyrosine kinase substrate p130cas (CAS) is a docking protein containing an SH3 domain near its N terminus, followed by a short proline-rich segment, a large central substrate domain composed of 15 repeats of the four amino acid sequence YxxP, a serine-rich region and a carboxy-terminal domain, which possesses consensus binding sites for the SH2 and SH3 domains of Src (YDYV and RPLPSPP, respectively). The SH3 domain of CAS mediates its interaction with several proteins involved in signaling pathways such as focal adhesion kinase (FAK), tyrosine phosphatases PTP1B and PTP-PEST, and the guanine nucleotide exchange factor C3G. As a homolog of the corresponding Src docking domain, the CAS SH3 domain binds to proline-rich sequences (PxxP) of its interacting partners that can adopt a polyproline type II helix. We have determined a high-resolution X-ray structure of the recombinant human CAS SH3 domain. The domain, residues 1 69, crystallized in two related space groups, P2(1) and C222(1), that provided diffraction data to 1.1 A and 2.1 A, respectively. The crystal structure shows, in addition to the conserved SH3 domain architecture, the way in which the CAS characteristic amino acids form an atypically charged ligand-binding surface. This arrangement provides a rationale for the unusual ligand recognition motif exhibited by the CAS SH3 domain. The structure enables modelling of the docking interactions to its ligands, for example from focal adhesion kinase, and supports structure-based drug design of inhibitors of the CAS-FAK interaction. PMID- 15784261 TI - Amyloid-like properties of bacterial inclusion bodies. AB - Bacterial inclusion bodies are major bottlenecks in protein production, narrowing the spectrum of relevant polypeptides obtained by recombinant DNA. While regarded as amorphous deposits formed by passive and rather unspecific precipitation of unfolded chains, we prove here that they are instead organized aggregates sharing important structural and biological features with amyloids. By using an Escherichia coli beta-galactosidase variant, we show that aggregation does not necessarily require unfolded polypeptide chains but rather depends on specific interactions between solvent-exposed hydrophobic stretches in partially structured species. In addition, purified inclusion bodies are efficient and highly selective nucleation seeds, promoting deposition of soluble homologous but not heterologous polypeptides in a dose-dependent manner. Finally, inclusion bodies bind amyloid-diagnostic dyes, which, jointly with Fourier transform infra red spectroscopy data, indicates a high level of organized intermolecular beta sheet structure. The evidences of amyloid-like structure of bacterial inclusion bodies, irrespective of potential applications in bioprocess engineering, prompts the use of bacterial models to explore the molecular determinants of protein aggregation by means of simple biological systems. PMID- 15784262 TI - Structural dynamics of the DnaK-peptide complex. AB - The molecular chaperone DnaK recognizes and binds substrate proteins via a stretch of seven amino acid residues that is usually only exposed in unfolded proteins. The binding kinetics are regulated by the nucleotide state of DnaK, which alternates between DnaK.ATP (fast exchange) and DnaK.ADP (slow exchange). These two forms cycle with a rate mainly determined by the ATPase activity of DnaK and nucleotide exchange. The different substrate binding properties of DnaK are mainly attributed to changes of the position and mobility of a helical region in the C-terminal peptide-binding domain, the so-called LID. It closes the peptide-binding pocket and thus makes peptide binding less dynamic in the ADP bound state, but does not (strongly) interact with peptides directly. Here, we address the question if nucleotide-dependent structural changes may be observed in the peptide-binding region that could also be connected to peptide binding kinetics and more importantly could induce structural changes in peptide stretches using the energy available from ATP hydrolysis. Model peptides containing two cysteine residues at varying positions were derived from the structurally well-documented peptide NRLLLTG and labelled with electron spin sensitive probes. Measurements of distances and mobilities of these spin labels by electron paramagnetic resonance spectroscopy (EPR) of free peptides or peptides bound to the ATP and ADP-state of DnaK, respectively, showed no significant changes of mobility nor distance of the two labels. This indicates that no structural changes that could be sensed by the probes at the position of central leucine residues located in the center of the binding region occur due to different nucleotide states. We conclude from these studies that the ATPase activity of DnaK is not connected to structural changes of the peptide-binding pocket but rather only has an effect on the LID domain or other further remote residues. PMID- 15784263 TI - Formation of native and non-native interactions in ensembles of denatured ACBP molecules from paramagnetic relaxation enhancement studies. AB - Paramagnetic relaxation enhancement measurements in the denatured state of ACBP have provided distance restraints that have been used in computer simulations to determine the conformational ensembles representing the denatured states of ACBP under a variety of conditions. A detailed comparison of the residual structure in the denatured state of ACBP under these different conditions has enabled us to infer that regions in the N and C-terminal parts of the protein sequence have a high tendency to interact in the unfolded state under physiological conditions. By comparing the structural features in the denatured states with those in the transition state for folding we also provided new insights into the mechanism of formation of the native state of this protein. PMID- 15784264 TI - Stabilization of the cold shock protein CspB from Bacillus subtilis by evolutionary optimization of Coulombic interactions. AB - The bacterial cold shock proteins (Csp) are used by both experimentalists and theoreticians as model systems for analyzing the Coulombic contributions to protein stability. We employ Proside, a method of directed evolution, to identify stabilized variants of Bs-CspB from Bacillus subtilis. Proside links the increased protease resistance of stabilized protein variants to the infectivity of a filamentous phage. Here, three cspB libraries were used for in vitro selections to explore the stabilizing potential of charged amino acids in Bs CspB. In the first library codons for nine selected surface residues were partially randomized, in the second one random mutations were introduced non specifically by error-prone PCR, and in the third one the spontaneous mutation rate of the phage in Escherichia coli was used. Stabilizing mutations were found at the surface positions 1, 3, 46, 48, 65, and 66. The contributions of these mutations to stability were characterized by analyzing them individually and in combination. The best combination (M1R, E3K, K65I, and E66L) increased the midpoint of thermal unfolding of Bs-CspB from 53.8 to 85.0 degrees C. The effects of most mutations are strongly context dependent. A good example is provided by the E3R mutation. It is strongly stabilizing (DeltaDeltaGD=11.1kJ mol(-1)) in the wild-type protein, but destabilizing (DeltaDeltaGD=-4.0kJ mol(-1)) in the A46K/S48R/E66L variant. The stabilizations by charge mutations did not correlate well with the corresponding changes in the protein net charge, and they could not be ascribed to the formation of ion pairs. Previous theoretical analyses did not identify the stabilization caused by the mutations at positions 1, 46, and 48. Also, electrostatics calculations based on protein net charge or charge asymmetry did not predict well the stability changes that occur when charged residues in Bs CspB are mutated. It remains a challenge to model the Coulombic interactions of charged residues in a protein and to determine their contributions to the Gibbs free energy of protein folding. PMID- 15784265 TI - The relationship between the flexibility of proteins and their conformational states on forming protein-protein complexes with an application to protein protein docking. AB - We investigate the extent to which the conformational fluctuations of proteins in solution reflect the conformational changes that they undergo when they form binary protein-protein complexes. To do this, we study a set of 41 proteins that form such complexes and whose three-dimensional structures are known, both bound in the complex and unbound. We carry out molecular dynamics simulations of each protein, starting from the unbound structure, and analyze the resulting conformational fluctuations in trajectories of 5 ns in length, comparing with the structure in the complex. It is found that fluctuations take some parts of the molecules into regions of conformational space close to the bound state (or give information about it), but at no point in the simulation does each protein as whole sample the complete bound state. Subsequent use of conformations from a clustered MD ensemble in rigid-body docking is nevertheless partially successful when compared to docking the unbound conformations, as long as the unbound conformations are themselves included with the MD conformations and the whole globally rescored. For one key example where sub-domain motion is present, a ribonuclease inhibitor, principal components analysis of the MD was applied and was also able to produce conformations for docking that gave enhanced results compared to the unbound. The most significant finding is that core interface residues show a tendency to be less mobile (by size of fluctuation or entropy) than the rest of the surface even when the other binding partner is absent, and conversely the peripheral interface residues are more mobile. This surprising result, consistent across up to 40 of the 41 proteins, suggests different roles for these regions in protein recognition and binding, and suggests ways that docking algorithms could be improved by treating these regions differently in the docking process. PMID- 15784266 TI - Modeling allosteric regulation of de novo pyrimidine biosynthesis in Escherichia coli. AB - With the emergence of multifaceted bioinformatics-derived data, it is becoming possible to merge biochemical and physiological information to develop a new level of understanding of the metabolic complexity of the cell. The biosynthetic pathway of de novo pyrimidine nucleotide metabolism is an essential capability of all free-living cells, and it occupies a pivotal position relative to metabolic processes that are involved in the macromolecular synthesis of DNA, RNA and proteins, as well as energy production and cell division. This regulatory network in all enteric bacteria involves genetic, allosteric, and physiological control systems that need to be integrated into a coordinated set of metabolic checks and balances. Allosterically regulated pathways constitute an exciting and challenging biosynthetic system to be approached from a mathematical perspective. However, to date, a mathematical model quantifying the contribution of allostery in controlling the dynamics of metabolic pathways has not been proposed. In this study, a direct, rigorous mathematical model of the de novo biosynthesis of pyrimidine nucleotides is presented. We corroborate the simulations with experimental data available in the literature and validate it with derepression experiments done in our laboratory. The model is able to faithfully represent the dynamic changes in the intracellular nucleotide pools that occur during metabolic transitions of the de novo pyrimidine biosynthetic pathway and represents a step forward in understanding the role of allosteric regulation in metabolic control. PMID- 15784267 TI - The complex effect of granulocyte colony-stimulating factor on human granulopoiesis analyzed by a new physiologically-based mathematical model. AB - Neutropenia, frequently a side effect of chemo- and radiotherapy, increases susceptibility to microbial infections and is a life-threatening condition. For realistically predicting drug treatment effects on granulopoiesis, we have constructed a new mathematical model of granulopoiesis in the bone marrow and in the peripheral blood, featuring cell cycle phase transition and detailed granulocyte-colony stimulating factor (G-CSF) pharmacokinetics (PK) and pharmacodynamics (PD), including intracellular second messenger. Using this model, in conjunction with clinical results, we evaluated the system parameters, implemented those in the model and successfully retrieved the results of several independent clinical experiments under a wide range of G-CSF regimens. Our results show that the introduction of G-CSF-controlled intracellular second messenger is indispensable for precise retrieval of the clinical results, and suggest that the half-life of this messenger varies between a single and multiple G-CSF administration schedules. In addition, our model provided reliable steady state, as well as dynamic, estimations of human granulopoiesis parameters. These included an estimation of apoptosis index in the post-mitotic compartment, which corroborates previous results. At present the model is used for suggesting improved drug regimens. PMID- 15784269 TI - An individual based model of rippling movement in a myxobacteria population. AB - Migrating cells of Myxococcus xanthus (MX) in the early stages of starvation induced development exhibit elaborate patterns of propagating waves. These so called rippling patterns are formed by two sets of waves travelling in opposite directions. It has been experimentally shown that formation of these waves is mediated by cell-cell contact signalling (C-signalling). Here, we develop an individual-based model to study the formation of rippling patterns in MX populations. Following the work of Igoshin et al. (Proc. Natl. Acad. Sci. 98 (2001) 14913) we consider each moving cell to have an internal clock which controls its turning behaviour and sensitivity to C-signal. Specifically, we examine the effects of changing: C-signal strength, sensitivity/refractoriness, cell density, and noise upon the formation and structure of the rippling patterns. We also consider three modified models that have no explicit refractory period and examine their ability to produce rippling patterns. PMID- 15784268 TI - The nk model and population genetics. AB - The nk model of fitness interactions is examined. This model has been used by previous authors to investigate the effects of fitness epistasis on substitution dynamics in molecular evolution, and to make broader claims about the importance of epistasis. To examine these claims, an infinite-allele approximation is introduced. In this limit, it is shown that the nk model is, at an appropriate level of description, formally identical to the non-epistatic House-of-Cards model--a well-studied model in theoretical population genetics. It is further shown that in many parameter regimes, the analytical results obtained from this infinite-allele approximation are very close to results from the full nk model (with a finite number of alleles per locus). The findings presented shed light on a number of previous results. PMID- 15784270 TI - Support vector machines for learning to identify the critical positions of a protein. AB - A method for identifying the positions in the amino acid sequence, which are critical for the catalytic activity of a protein using support vector machines (SVMs) is introduced and analysed. SVMs are supported by an efficient learning algorithm and can utilize some prior knowledge about the structure of the problem. The amino acid sequences of the variants of a protein, created by inducing mutations, along with their fitness are required as input data by the method to predict its critical positions. To investigate the performance of this algorithm, variants of the beta-lactamase enzyme were created in silico using simulations of both mutagenesis and recombination protocols. Results from literature on beta-lactamase were used to test the accuracy of this method. It was also compared with the results from a simple search algorithm. The algorithm was also shown to be able to predict critical positions that can tolerate two different amino acids and retain function. PMID- 15784271 TI - Modeling of slow glutamate diffusion and AMPA receptor activation in the cerebellar glomerulus. AB - Synaptic conductances are influenced markedly by the geometry of the space surrounding the synapse since the transient glutamate concentration in the synaptic cleft is determined by this geometry. Our paper is an attempt to understand the reasons for slow glutamate diffusion in the cerebellar glomerulus, a structure situated around the enlarged mossy fiber terminal in the cerebellum and surrounded by a glial sheath. For this purpose, analytical expressions for glutamate diffusion in the glomerulus were considered in models with two-, three , and fractional two-three-dimensional (2D-3D) geometry with an absorbing boundary. The time course of average glutamate concentration in the synaptic cleft of the mossy fiber-granule cell connection was calculated for both direct release of glutamate from the same synaptic unit, and for cumulative spillover of glutamate from neighboring release sites. Several kinetic schemes were examined, and the parameters of the diffusion models were estimated by identifying theoretical activation of AMPA receptors with direct release and spillover components of published experimental AMPA receptor-mediated EPSCs. For model selection, the correspondence of simulated paired-pulse ratio and EPSC increase after prevention of desensitization to experimental values were also taken into consideration. Our results suggest at least a 7- to 10-fold lower apparent diffusion coefficient of glutamate in the porous medium of the glomerulus than in water. The modeling of glutamate diffusion in the 2D-3D geometry gives the best fit of experimental EPSCs. We show that it could be only partly explained by normal diffusion of glutamate in the complex geometry of the glomerulus. We assume that anomalous diffusion of glutamate occurs in the glomerulus. A good match of experimental estimations and theoretical parameters, obtained in the simulations that use an approximation of anomalous diffusion by a solution for fractional Brownian motion, confirms our assumption. PMID- 15784272 TI - Modelling genetic networks with noisy and varied experimental data: the circadian clock in Arabidopsis thaliana. AB - Circadian clocks in all organisms include feedback loops that generate rhythmic expression of key genes. We model the first such loop proposed for the clock of Arabidopsis thaliana, the experimental model species for circadian timing in higher plants. As for many biological systems, there are no experimental values for the parameters in our model, and the data available for parameter fitting is noisy and varied. To tackle this we constructed a cost function, which quantifies the agreement between our model and various key experimental features. We then undertook an efficient global search of parameter space, to test whether the proposed circuit can fit the experimental data. Using this approach we show that circadian clock models can function well with low cooperativity in transcriptional regulation, whereas high cooperativity has been a feature of previous (hand-fitted) clock models in other species. Our optimized solution for the Arabidopsis clock model fits several, but not all, of the key experimental features. We test the predicted effects of well-characterized mutations in the clock circuit and show the phases of the circadian cycle where additional components that are yet to be identified experimentally must be present to complete the circadian feedback loop. PMID- 15784273 TI - Detecting differentially expressed genes by relative entropy. AB - DNA microarray experiments have generated large amount of gene expression measurements across different conditions. One crucial step in the analysis of these data is to detect differentially expressed genes. Some parametric methods, including the two-sample t-test (T-test) and variations of it, have been used. Alternatively, a class of non-parametric algorithms, such as the Wilcoxon rank sum test (WRST), significance analysis of microarrays (SAM) of Tusher et al. (2001), the empirical Bayesian (EB) method of Efron et al. (2001), etc., have been proposed. Most available popular methods are based on t-statistic. Due to the quality of the statistic that they used to describe the difference between groups of data, there are situations when these methods are inefficient, especially when the data follows multi-modal distributions. For example, some genes may display different expression patterns in the same cell type, say, tumor or normal, to form some subtypes. Most available methods are likely to miss these genes. We developed a new non-parametric method for selecting differentially expressed genes by relative entropy, called SDEGRE, to detect differentially expressed genes by combining relative entropy and kernel density estimation, which can detect all types of differences between two groups of samples. The significance of whether a gene is differentially expressed or not can be estimated by resampling-based permutations. We illustrate our method on two data sets from Golub et al. (1999) and Alon et al. (1999). Comparing the results with those of the T-test, the WRST and the SAM, we identified novel differentially expressed genes which are of biological significance through previous biological studies while they were not detected by the other three methods. The results also show that the genes selected by SDEGRE have a better capability to distinguish the two cell types. PMID- 15784274 TI - Models of active transport of ions in biomembranes of various types of cells. AB - Nonequilibrium statistical models of the active transport of ions in biomembranes have been constructed. Differences of chemical potentials of the ATP-ADP reaction and the electrochemical potential of ions were taken as the thermodynamic forces responsible for the flow of ions through the membrane. The active transport of ions was viewed as a cross phenomenon arising from the chemical reaction of the ATP hydrolysis. These models provide independent calculations of the resting potential at the biomembrane and concentrations of ions in a cell on the assumption the free energy of the ATP-ADP reaction is fully (without the dissipation loss) converted to the free energy of transported ions. They take into account the presence of nonpenetrating ions in a cell. It was shown that different concentrations of nonpenetrating ions have a considerable effect on the resting potential. The proposed models were compared with experimental data obtained for different types of cells including neurons, muscular cells, bacteria, plants, and mitochondria. Calculated values of the membrane potential and ion concentrations were in good qualitative agreement with experimental data. PMID- 15784275 TI - Modelling development of epidemics with dynamic small-world networks. AB - We discuss the dynamics of a minimal model for spreading of infectious diseases, such as various types of influenza. The spreading takes place on a dynamic small world network and can be viewed as comprising short- and long-range spreading processes. We derive approximate equations for the epidemic threshold as well as the spreading dynamics, and show that there is a good agreement with numerical discrete time-step simulations. We then analyse the dependence of the epidemic saturation time on the initial conditions, and outline a possible method of utilizing the model in predicting the development of epidemics based on early figures of infected. Finally, we compare time series calculated with our model to real-world data. PMID- 15784276 TI - Response to temporal parameter fluctuations in biochemical networks. AB - Metabolic response coefficients describe how variables in metabolic systems, like steady state concentrations, respond to small changes of kinetic parameters. To extend this concept to temporal parameter fluctuations, we define spectral response coefficients that relate Fourier components of concentrations and fluxes to Fourier components of the underlying parameters. It is also straightforward to generalize other concepts from metabolic control theory, such as control coefficients with their summation and connectivity theorems. The first-order response coefficients describe forced oscillations caused by small harmonic oscillations of single parameters: they depend on the driving frequency and comprise the phases and amplitudes of the concentrations and fluxes. Close to a Hopf bifurcation, resonance can occur: as an example, we study the spectral densities of concentration fluctuations arising from the stochastic nature of chemical reactions. Second-order response coefficients describe how perturbations of different frequencies interact by mode coupling, yielding higher harmonics in the metabolic response. The temporal response to small parameter fluctuations can be computed by Fourier synthesis. For a model of glycolysis, this approximation remains fairly accurate even for large relative fluctuations of the parameters. PMID- 15784277 TI - Mathematical analysis demonstrates that interferons-beta and -gamma interact in a multiplicative manner to disrupt herpes simplex virus replication. AB - Several studies suggest that the innate interferons (IFNs), IFN-alpha and IFN beta, can act in concert with IFN-gamma to synergistically inhibit the replication of cytomegalovirus and herpes simplex virus type 1 (HSV-1). The significance of this observation is not yet agreed upon in large part because the nature and magnitude of the interaction between IFN-alpha/beta and IFN-gamma is not well defined. In the current study, we resolve this issue by demonstrating three points. First, the hyperbolic tangent function, tanh (x), can be used to describe the individual effects of IFN-beta or IFN-gamma on HSV-1 replication over a 320,000-fold range of IFN concentration. Second, pharmacological methods prove that IFN-beta and IFN-gamma interact in a greater-than-additive manner to inhibit HSV-1 replication. Finally, the potency with which combinations of IFN beta and IFN-gamma inhibit HSV-1 replication is accurately predicted by multiplying the individual inhibitory effects of each cytokine. Thus, IFN-beta and IFN-gamma interact in a multiplicative manner. We infer that a primary antiviral function of IFN-gamma lies in its capacity to multiply the potency with which IFN-alpha/beta restricts HSV-1 replication in vivo. This hypothesis has important ramifications for understanding how T lymphocyte-secreted cytokines such as IFN-gamma can force herpesviruses into a latent state without destroying the neurons or leukocytes that continue to harbor these viral infections for the lifetime of the host. PMID- 15784278 TI - NADPH-dependent metabolism of 17beta-estradiol and estrone to polar and nonpolar metabolites by human tissues and cytochrome P450 isoforms. AB - The endogenous estrogens, 17beta-estradiol (E(2)) and estrone (E(1)), undergo extensive metabolism in animals and humans, and a large number of their hydroxylated and keto metabolites have been identified in biological samples. The formation of most of the oxidative estrogen metabolites is catalyzed by cytochrome P450 (CYP) enzymes. Precise knowledge of the CYP-mediated formation of these metabolites, particularly those with unique biological activities (e.g., 4 hydroxy-E(2), 16alpha-hydroxy-E(1), 15alpha-hydroxy-E(2), 16-epiestriol, and 2 methoxyestradiol) in human liver and extrahepatic target tissues and cells, would add significantly to our understanding of the diverse biological functions that are associated with endogenous estrogens. In this article, we review recent results on the NADPH-dependent metabolism of endogenous estrogens to polar (hydroxylated and keto) metabolites as well as to nonpolar metabolites by human tissues and recombinant human CYP isoforms. The available data show that a large number of polar and nonpolar metabolites of E(2) and E(1) are formed by human tissues, and a variety of human CYP isoforms are involved in the NADPH-dependent formation of polar as well as nonpolar estrogen metabolites. These enzymes have varying degrees of catalytic activity and distinct regioselectivity. PMID- 15784279 TI - Epoxidation and reduction of cholesterol, 1,4,6-cholestatrien-3-one and 4,6 cholestadien-3beta-ol. AB - Many naturally occurring polyhydroxylated sterols and oxysterols exhibit potent biologic activities. This paper describes reagent and position selectivity of epoxidation and reduction of cholesterol derivatives. Cholesterol was reacted with m-chloroperoxybenzoic acid (m-CPBA) to form 5alpha,6alpha-epoxycholestan 3beta-ol, but in reaction with 30% H(2)O(2), it did not reacted. 1,4,6 cholestatrien-3-one was obtained from cholesterol and 2,3-dichloro-5,6-dicyano 1,4-benzoquinone in dioxane. 1,4,6-cholestatrien-3-one was reacted with 30% H(2)O(2) and 5% NaOH in methanol to give 1alpha,2alpha-epoxy-4,6-cholestadien-3 one, which was stereoselectively reduced with NaBH(4) to form 1alpha,2alpha-epoxy 4,6-cholestadien-3beta-ol and reduced with Li metal in absolute ethanol to give 2 ethoxy-1,4,6-cholestatrien-3-one. And 1,4,6-cholestatrien-3-one was epoxidized with m-CPBA in dichloromethane to afford 6alpha,7alpha-epoxy-1,4-cholestadien-3 one, which was reacted with NaBH(4) to synthesize 6alpha-hydroxy-4-cholesten-3 one and reduced Li metal in absolute ethanol to form 2-ethoxy-1,4,6-cholestatrien 3-one, respectively. 1,4,6-cholestatrien-3-one was reduced with NaBH(4) in absolute ethanol to form 4,6-cholestadien-3beta-ol, which was reacted with 30% H(2)O(2) to leave original compound, but was reacted with m-CPBA to give 4beta,5beta-epoxy-6-cholesten-3beta-ol as the major product and 4beta,5beta-epoxy 6alpha,7alpha-epoxycholestan-3beta-ol as the minor product. PMID- 15784280 TI - Novel action of estrone on vascular tissue: regulation of NOS and COX activity. AB - The hypothesis tested in the present work is that estrone non-genomically regulates aortic nitric oxide synthase (NOS) and cyclooxygenase (COX) activities in female rats, and that such regulation depends on ovarian function. We found that physiological concentrations of estrone (E(1)) (0.1-10nM) significantly increased nitric oxide (NO) production (133 and 163% above control). The stimulatory action of E(1) on NOS activity was independent of calcium influx since the increase in NO elicited by the hormone was not affected by EGTA or verapamil. When COX activity was measured, we observed that estrone enhanced thromboxane (TXB(2)) production and prostacyclin (PGI(2)) release, but not prostaglandin (PGF(2), PGD(2), and PGE(2)) synthesis. Finally we demonstrated that the hormonal effect on NOS activity was not detected in rat aortic strips (RAS) isolated from animals deprived of ovarian activity (FR(-)) or ovariectomized rats (OVX). These results suggest that estrone exerts a direct, non-genomic action on rat aortic metabolism, which involves NOS and COX activation and depends on ovarian activity. PMID- 15784281 TI - New glycosides of the campesterol derivative from the rhizomes of Tacca chantrieri. AB - Seven new glycosides of the campesterol derivative (24R,25S)-ergost-5-ene 3beta,26-diol (1-7) were isolated from the rhizomes of Tacca chantrieri (Taccaceae). Their structures were determined by extensive spectroscopic analysis, including 2D NMR data, and a few chemical transformations. PMID- 15784282 TI - MRI quantification in vivo of corticosteroid induced thymus involution in mice: correlation with ex vivo measurements. AB - Thymus involution is a useful marker of transactivation-mediated side effects in preclinical therapeutic index testing of new anti-inflammatory glucocorticosteroids, and is usually measured post mortem. We have validated the use of MRI for non-invasive in vivo measurement of mouse thymus involution induced by dexamethasone (DEX). Tl-weighted spin echo 7 T images provided satisfactory contrast between thymus and surrounding connective tissue and fat. Increasing doses of DEX caused thymus involution, reflected in MRI volume (87+/ 14, 33+/-10, 28+/-6, 16+/-7 microl in dosage groups of Cremophor vehicle, 1, 10 and 30 mg/kg subcutaneous respectively, n=6/group, mean+/-standard deviation) and post mortem wet weight (64+/-12, 33+/-6, 25+/-9, 23+/-8 mg). Correlation between MRI volumes and wet weights was very good (r=0.842). Measuring pre-dose MRI volumes and then assessing DEX effects as post-dose change from baseline produced no statistical advantage relative to considering post-dose MRI thymus volume alone, probably due to variability in pre-dose baseline values compounding post dose variability. Smaller group sizes were sufficient to achieve a given statistical power using MRI post-dose volume than using wet weight, suggesting a role for MRI in differentiating the effects of compounds which produce similar effects, or in contexts where the use of large groups of animals is impractical or ethically unacceptable. PMID- 15784284 TI - Cholinesterase inhibitory pregnane-type steroidal alkaloids from Sarcococca hookeriana. AB - The bioassay-guided phytochemical investigation on Sarcococca hookeriana have resulted in the isolation of four new pregnane-type steriodal alkaloids: hookerianamide-D [(2'E,20S)-20-(N,N-formyl(methyl)amino)-3beta-(3',4'-dimethyl-2' pentenamido)-5alpha-pregnane] (1), hookerianamide-E [(2'E,20S)-20-(N,N dimethylamino)-3beta-(senecioylamino)-5alpha-pregn-14-en-2beta-O-acetate] (2), hookerianamide-F [(2'E,20S)-20-(N-methylamino)-3beta-(tigloylamino)-5alpha-pregn 2,14-dien-4-one] (3), and hookerianamide-G [(20S)-20-(N,N-dimethylamino)-3beta-(N methylbenzamido)-5alpha-pregn-4beta-O-acetate] (4), along with five known alkaloids 5-9. Their structures were determined by spectroscopic analysis. These steroidal alkaloids and chemically derived derivatives of compound 5 have displayed varying degree of inhibitory activities against acetylcholinesterase and butyrylcholinesterase enzymes in a concentration-dependent fashion, with the IC(50) values ranging from 1.5 to 148.2 and 0.6 to 100.2 microM, respectively. PMID- 15784283 TI - Generation of a single-chain Fv fragment for the monitoring of deoxycholic acid residues anchored on endogenous proteins. AB - A subset of lipophillic bile acids, including deoxycholic acid (DCA) and lithocholic acid (LCA), are thought to be biologically transformed into reactive intermediates forming covalently modified, "tissue-bound" bile acids that can exert several toxic effects. We have generated a single-chain Fv fragment (scFv) as a probe to monitor DCA residues anchored on proteins. DNA fragments encoding the variable heavy (V(H)) and light (V(L)) domains of a mouse antibody raised against a DCA hapten (Ab #88) were cloned by rapid amplification of cDNA 5'-ends. These sequences were combined via a common linker sequence coding (Gly(4)Ser)(3) to construct a single scFv gene with the gene segments in the following order: 5' V(H)-linker-V(L)-3'. This construct was subcloned into an antibody-expression vector, pEXmide 5; soluble scFv protein was then expressed in the bacterial periplasm of the XLOLR Escherichia coli strain. In a competitive enzyme-linked immunosorbent assay using DCA-coated microtiter plates, the scFv provided a dose response curve for free DCA ranging between 2 and 5000 pg/assay. The scFv reacts similarly with the l-lysine adduct of DCA (cross-reactivity, 72%), while bile acids having a modified DCA steroid skeleton were well-discriminated (cross reactivity, <1%). This scFv could also monitor trace amounts of DCA residues anchored on a protein through DCA acyl adenylate reactions, the likely reactive intermediate. The present scFv may be a useful tool for trace characterization of tissue-bound bile acids; this usefulness may be significantly enhanced by fusion with signal-generating proteins, such as alkaline phosphatase or green fluorescent protein. PMID- 15784285 TI - Discovery, biosynthesis, and structure elucidation of new metabolites of norandrostenedione using in vitro systems. AB - The aim of our study was to demonstrate the positive impact that in vitro systems could have on the synthesis and characterization of unknown metabolites of banned doping agents. Using norandrostenedione (estr-4-en-3,17-dione), we were able to identify and characterize by GC/MS and LC/UV/MS several new hydroxylated metabolites formed in human hepatocyte incubations. The site of hydroxylation of M1, M2, M3, and M5 was demonstrated to be at C-6beta position by incubating estr 4-en-6beta-ol-3,17-dione (M4), which is the direct 6beta-hydroxylated metabolite of norandrostenedione. The structure of M5 was confirmed to be estr-4-en 6beta,17beta-diol-3-one (6beta-hydroxynortestosterone) using a commercially available authentic standard. For the other metabolites, M1, M2, and M3, no standards were available. Due to limited access to fresh human liver tissues, in vitro incubation conditions in rat liver subcellular fractions and hepatocytes were optimized as an alternative to produce sufficient quantities of the unknown metabolites for MS and/or NMR characterization. The structure of M1 was assigned to 5alpha-estran-3alpha,6beta-diol-17-one (6beta-hydroxynorandrosterone) and M3 to 5alpha-estran-3beta,6beta-diol-17-one (6beta-hydroxynorepiandrosterone) based on NMR data. M2 is proposed to be 5beta-estran-3alpha,6beta-diol-17-one (6beta hydroxynoretiocholanolone) based on GC/MS fragmentation of the TMS-enol bis-TMS ether derivative. The in vitro approach reported here, in addition to urinary excretion studies in humans, could contribute significantly to the discovery, the synthesis, and structure elucidation of new markers of doping agents. PMID- 15784286 TI - Neurosteroids: metabolism in human intestine microsomes. AB - Both dehydroepiandrosterone (DHEA) and epiandrosterone (EpiA) are substrate for cytochrome P450 species and enzymes that produce 7alpha- and 7beta-hydroxylated metabolites in the brain and other organs. In contrast to DHEA and EpiA, the 7 hydroxylated derivatives were shown to mediate neuroprotection, and 7beta-hydroxy EpiA was the most potent. The suggested use of any of these steroids as drugs administered per os for neuroprotection requires the assessment of their metabolism in the human intestine and liver. To achieve this, we produced radio labeled 7alpha-hydroxy-DHEA, 7beta-hydroxy-DHEA, 7alpha-hydroxy-EpiA and 7beta hydroxy-EpiA that were used as substrates in incubations with human intestine microsomes supplemented with reduced or oxidized cofactors. Identity of the radio labeled metabolites obtained was determined by gas chromatography/mass spectrometry after comparison with authentic steroid references. The proportions of metabolites produced resulted from their radioactivity contents. The only metabolite obtained with DHEA, EpiA, 7alpha-hydroxy-DHEA and 7beta-hydroxy-DHEA substrates was its 17beta-reduced derivative, thus inferring the presence of 17beta-hydroxysteroid oxidoreductases in the human intestine microsomes. In addition to the 7alpha-hydroxy-EpiA and 7beta-hydroxy-EpiA substrates, their 17beta-reduced metabolites were obtained with 7beta-hydroxy-EpiA and 7alpha hydroxy-EpiA, respectively. The identity of the enzyme responsible for the 7alpha hydroxy-EpiA/7beta-hydroxy-EpiA inter-conversion is unknown. The incubation conditions used produced these metabolites in low but significant yields that suggest their presence in the portal blood before access to the liver. PMID- 15784287 TI - Studies on Bacillus stearothermophilus. Part IV. Influence of enhancers on biotransformation of testosterone. AB - The impact of chemical enhancers on the biotransformation of testosterone has been exploited. Application of crude cell concentrates to produce Bacillus stearothermophilus-mediated bioconversion of testosterone at 65 degrees C for 72 h has been examined. After incubation, the xenobiotic substrate was added to the concentrated whole cell suspensions. The enhancer molecules were included in the whole cell suspension. The resultant products, after extraction into an organic solvent, were purified by thin layer chromatography and identification was carried out through spectroscopic data. Five steroid metabolites 9,10-seco-4 androstene-3,9,17-trione, 5alpha-androstan-3,6,17-trione, 17beta-hydroxy-5alpha androstan-3,6-dione, 3beta,17beta-dihydroxyandrost-4-ene-6-one and 17beta hydroxyandrost-4,6-diene-3-one were identified as biotransformation products of testosterone. A possible biosynthetic route for these bioconversion products is postulated. PMID- 15784288 TI - Glucocorticoid receptors and physiology: a personal history. PMID- 15784289 TI - Immunostimulatory oligodeoxynucleotides induce dolphin neutrophil NADPH-oxidase activation in a CpG-independent but phosphorothioate backbone-dependent manner. AB - Immunocytes, which include antigen-presenting cells, B cells, natural killer cells and neutrophils, can be stimulated directly or indirectly with bacterial DNA and synthetic oligodeoxynucleotides (ODNs) with different structures and sequences. In the present study, we investigated the effect of synthetic ODNs on the respiratory burst of dolphin neutrophils using a chemiluminescence assay. Phosphorothioate (PS)-ODNs dose-dependently induced the respiratory burst, while phosphodiester (PO)-ODNs did not, regardless of CpG-content. The PS-ODN-induced activity was completely abolished by the flavoprotein inhibitor diphenyleneiodonium, which indicates that the NADPH-oxidase is activated by PS ODNs. These results reveal that PS-ODNs induce dolphin neutrophil NADPH-oxidase activation in a CpG motif-independent but phosphorothioate-dependent manner. PMID- 15784290 TI - Antimicrobial peptide defenses against chytridiomycosis, an emerging infectious disease of amphibian populations. AB - Chytridiomycosis, an emerging infectious disease (EID) of the skin caused by the chytrid fungus, Batrachochytrium dendrobatidis, has been linked with continuing amphibian population declines in the western USA, Central America, Europe, Africa, and Australia. Genetic analysis suggests that B. dendrobatidis is a recently emerged pathogen. This article reviews the biology of this pathogenic chytrid and the evidence for chytridiomycosis as a cause of declines in amphibian populations worldwide. Data are presented to show that antimicrobial peptides, produced in granular glands of the skin and released in high concentrations into skin secretions, are highly effective in inhibiting growth of B. dendrobatidis in vitro and may provide limited protection for some species. Ongoing studies suggest a correlation between resistance to lethal infection by B. dendrobatidis and synthesis of antimicrobial peptides by the host amphibian, but further research is needed to define better the role of antimicrobial peptides in protection of amphibian populations and the effect of environmental factors upon antimicrobial peptide synthesis. PMID- 15784291 TI - Monoclonal antibodies identify four distinct annelid leukocyte markers. AB - This paper describes for the first time the production and characterization of a library of monoclonal antibodies (anti-EFCC clones) raised against coelomocyte (leukocyte) markers of Eisenia fetida earthworm. Leukocyte subgroups are components of earthworm innate immunity that require a more precise characterization using immunological markers. Flow cytometry, immunocytochemistry, and immunoprecipitation analyzed and confirmed the specificity of anti-EFCC clones. Anti-EFCC mAbs revealed different leukocyte subpopulations and various staining patterns on tissues. Two functional assays (e.g. phagocytosis and encapsulation) further characterized EFCC clusters revealing a common coelomocyte marker and three subpopulation-specific markers. No crossreactivity was found on human, mouse, rat or cells from Drosophila melanogaster but immunoreactivity was detected on snail (Planorbarius corneus) tissues. Immunohistochemical results suggest mesodermal origin of all coelomocyte subgroups that agree with classical morphological analyses. PMID- 15784292 TI - Expression of genes related to the early immune response in rainbow trout (Oncorhynchus mykiss) after viral haemorrhagic septicemia virus (VHSV) infection. AB - In the last few years, many cytokine and other immune related genes have been identified in different teleost species, thus allowing their study at a molecular level. However, very little is known about their effect on fish antiviral responses. In the current work, we have studied the effect of viral haemorrhagic septicemia virus (VHSV) infection on the expression of different immune genes in rainbow trout (Oncorhynchus mykiss) through semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). We have studied the effect of the viral infection on the expression of different cytokines such as interleukin 1beta (IL-1beta) and transforming growth factor beta (TGF-beta), the CXC chemokine IL-8, and other immune genes such as inducible nitric oxide synthase (iNOS) and the class II major histocompatibility complex (MHC II). The virus induced an increased transcription of IL-1beta in the spleen, and to a lesser extent in the head kidney and liver at early times post-infection. IL-8 transcription was also significantly induced with the virus in the spleen at early times post-infection. TGF-beta transcription was significantly induced in VHSV infection in the spleen and liver. In the spleen, a significant induction of TGF-beta at day 1 post-infection was observed. A further significant increase occurred in the spleen and liver at day 7 post-infection. No effect of the virus on MHC II expression was ever observed while iNOS was induced in the spleen, head kidney and liver of VHSV-infected fish mostly at day 7 post-infection. These results constitute a first step towards the understanding of which molecules may have a role in antiviral defence in fish. PMID- 15784293 TI - Poly I:C induces an antiviral state against Ictalurid Herpesvirus 1 and Mx1 transcription in the channel catfish (Ictalurus punctatus). AB - In vivo studies were carried out to investigate the protective effect of the interferon inducer poly I:C against channel catfish virus (CCV). Channel catfish were stimulated by intraperitoneal injection of 50 microg of poly I:C or PBS at various days prior to immersion challenge with CCV. Mortality in the poly I:C group was significantly reduced from 70% to 3% at day 1 compared to the PBS controls. Mortality increased at day 3 but was still significantly different from the PBS controls. Mx1 transcription was significantly higher only at day 1. In an additional study Mx1 transcription was monitored in the liver, kidney, gills, spleen, and intestine at various time points post-stimulation with either poly I:C or CCV. Mx1 mRNA was significantly elevated in all organs only at day 1 post injection with poly I:C. In response to CCV, Mx1 transcription was not significantly elevated until day 3 post-challenge, but remained elevated in certain organs until day 7. PMID- 15784294 TI - Molecular characterization of the nonspecific cytotoxic cell receptor (NCCRP-1) demonstrates gilthead seabream NCC heterogeneity. AB - Teleost fish NCCs (nonspecific cytotoxic cells) are thought to be the evolutionary precursors of the mammalian NK cells. A novel mechanism mediating the NCC-mediated cytotoxicity has been described in teleosts. Now, this NCC receptor protein-1 (NCCRP-1) was characterized in gilthead seabream. The NCCRP-1 full-length sequence contains 1036 bp with an open reading frame of 702 bp. A comparison of the predicted 233-amino acid protein with several fish orthologues indicates a highly conserved sequence containing the F-box associated (FBA) domain and proline-rich motifs (PRM) characteristics of this family. The phylogenetical tree shows that seabream NCCRP-1 belongs to the NCCRP subfamily within the FBA family of proteins. This is a single copy gene with a constitutive and ubiquitous expression as determined by RT-PCR and flow cytometry. The results show that lymphocytes, monocyte/macrophages and acidophilic granulocytes from lymphoid tissues express the receptor, both at gene and protein level. Immunofluorescence microscopic observations confirm the previous results. The implications of this receptor on seabream NCC activity are discussed. PMID- 15784296 TI - Avian thymic hormone treatment of peripheral blood mononuclear cells from young chicks stimulates acute graft-versus-host reaction in chicken embryos. AB - Avian thymic hormone (ATH) is a parvalbumin produced by epithelial cells in the thymic cortex of chickens and circulates in the blood on a 5-day cycle. It stimulates precocious development of cell-mediated immunity. The effect of partially purified extracts of thymus (TE) and purified ATH were tested for their effect on the acute graft-versus-host reaction (GVHR). Treatment of chicks for their first 3-days of life did not enhance the acute GVHR produced by their PBMC in 14-day-old embryos. PBMC from 3-day-old chicks were treated in vitro with TE, ATH, thymosin fraction 5 or thymosin alpha1 for 2 h and injected into 14-day-old embryos. Bone marrow cells and thymic lymphocytes were treated with TE. Only PBMC treated with TE or ATH produced an enhanced acute GVHR. Because ATH targets gammadelta T cells, the data implicate participation of donor gammadelta T cells in the acute GVHR. PMID- 15784295 TI - Developmentally programmed expression of AID in chicken B cells. AB - In mice, activation induced deaminase, AID, is expressed only in germinal center B cells. It is required for the initiation of somatic hypermutation and class switch recombination. In chickens and most mammals immunoglobulin gene rearrangement generates limited diversity and the primary immunoglobulin repertoire depends on subsequent somatic hypermutation or gene conversion. Immunoglobulin gene conversion in chickens starts in the embryonic bursa, before antigen exposure. The demonstrated requirement for AID for gene conversion in the bursal lymphoma cell line, DT40, implies developmental regulation of AID expression. To test this prediction, we examined the timing and location of AID mRNA expression. An abrupt increase in AID mRNA coincided with the onset of extensive Ig gene conversion in the bursa. Expression was also detected at earlier stages, implying either that expression of AID is not the only controlling factor for gene conversion, or that gene conversion can precede the formation of bursal follicles. PMID- 15784297 TI - The role of vena caval filters in the management of venous thromboembolism. AB - Venous thromboembolism (VTE) is a significant cause of morbidity and mortality worldwide. Anticoagulation is the preferred treatment for VTE, however, in selected patient populations, the risk of bleeding from anticoagulation outweighs its benefit. For these patients, alternative methods of pulmonary embolism (PE) prevention are needed. In 1865, Armand Trousseau proposed the concept of vena caval interruption. This idea has evolved from surgical ligation of the inferior vena cava (IVC) to the development of percutaneous IVC filters, and most recently, retrievable filters. With these advances in technology, there has been a dramatic increase in the use of IVC filters. Despite the vast number of reports on the use of IVC filters, there are limited data on their efficacy in many clinical circumstances. In this review, we will discuss the currently available IVC filters, data on their efficacy and safety and our assessment of appropriate indications for their use. PMID- 15784298 TI - Non-pharmacologic approaches to graft-versus-host prevention. AB - Allogeneic transplant offers a curative option for selected hematological malignancies and improved disease free survival in those where cure is not possible. For patients in these categories, Graft-versus-host disease (GVHD) occurs often and is a significant cause of morbidity and mortality following transplantation. Pharmacological immunosuppression has considerable toxicity and controls GVHD in about 75% of transplant recipients when used in combination and continually. Thus, there is the need for viable and non-toxic alternatives for GVHD prevention and this is the impetus for seeking novel non-pharmacological strategies. Most techniques directed to this end are based on various permutations of T-cell depletion. Total T-cell depletion was the first graft engineering strategy employed but while beneficial, it was associated with delay in engraftment and early relapses. Modern depletion and adoptive immunotherapy techniques are directed toward specific subsets of T-cells which include alloreactive, naive, NK cells, T-helper and cytotoxic T-cells. While large bodies of preclinical evidence attest to the efficacy of currently available non pharmacological prevention techniques, such is not the case with vigorous clinical trials. Where available, the proven non-pharmacological prevention techniques are yet to be fully interwoven into the patient care arena. Integration of available strategies and continuing research are therefore acutely needed to improve the chronic morbidity and mortality seen with both allogeneic transplant-associated GVHD and the use of pharmaceutical immunosuppressants. PMID- 15784299 TI - Bcl-2 antisense therapy in B-cell malignancies. AB - Bcl-2 is an apoptosis regulating protein, overexpression of which is associated with chemotherapy resistant disease, aggressive clinical course, and poor survival in patients with B-cell lymphoproliferative disorders. Overexpression of Bcl-2 protein results in an aberrant intrinsic apoptotic pathway that confers a protective effect on malignant cells against a death signal (e.g., chemotherapy or radiotherapy). Downregulation of this oncoprotein, thus, represents a possible new way to target clinically aggressive disease. Preclinical studies have shown that this oncoprotein can be effectively decreased by Bcl-2 antisense in malignant lymphoid cells and can reverse chemotherapy resistance, as well as enhance the anti-apoptotic potential of both chemotherapeutic and biologic agents. Ongoing clinical trials are exploring the role of Bcl-2 downregulation with oblimersen (Bcl-2 antisense) in patients with non-Hodgkin's lymphoma, chronic lymphocytic leukemia and multiple myeloma. Early results from these studies are promising and support the proof of the principle. As these studies are completed and mature data emerges, the role of Bcl-2 antisense therapy in the treatment of B-cell malignancies will become clearer. PMID- 15784300 TI - Microarray-based gene expression profiling of hematologic malignancies: basic concepts and clinical applications. AB - Each cell in our body contains a set of tens of thousands of genes, out of which a set of several thousands determines the cell's characteristics. The deciphering of the sequence of the human genome combined with the technical feasibility to simultaneously measure the gene expression levels of thousands of genes had revolutionized our understanding of cellular processes. This ability has great significance in our comprehension of the mechanisms that bring about diseases in general and hematologic malignancies in particular. Several new high-throughput technologies, commonly referred as microarrays, enable us to perform such measurements and concurrently, bioinformatic and statistical tools were developed to analyze the data obtained by using microarrays. In this review we present examples of analyses of hematologic malignancies using microarrays which contribute to refinement of diagnosis, identification of novel disease subtypes and of relationships between diseases that were previously considered to be unrelated, prediction of response to treatment and identification of genes and pathways linked to pathogenesis, thus defining targets to rational therapy. PMID- 15784301 TI - Plasticity of synaptic transmission in autonomic ganglia. AB - Synaptic plasticity is a term that describes long-lasting changes in the efficacy of synaptic transmission resulting from certain patterned activities of the presynaptic nerve. One form of synaptic plasticity, long-term potentiation (LTP), is an activity-dependent marked increase in synaptic efficacy that has been extensively studied in various regions of the central nervous system, particularly the hippocampus, where LTP is widely believed to be a cellular correlate of learning and memory. A similar phenomenon has been identified in sympathetic ganglia even before Bliss and Lomo coined the term LTP in 1973. Ganglionic LTP (gLTP) of the nicotinic pathway is a similarly long-lasting increase in synaptic effectiveness that can be induced in ganglia following a brief train of relatively high frequency stimulation (HFS) of the preganglionic nerve. Remarkably similar to the LTP of the hippocampus, gLTP has been demonstrated in autonomic ganglia from a number of vertebrates including mammalian, amphibian and avian species. Several other forms of long-lasting increases in synaptic effectiveness have been demonstrated in sympathetic ganglia following exposure to adrenergic agonists, neuroactive peptides and cyclic nucleotides and even after a challenge by an antigen. The main emphasis of this review, however, will be on the activity-dependent gLTP of the mammalian sympathetic ganglia, in particular the superior cervical ganglion of the rat. Since the last, excellent and comprehensive review of this ganglionic function by Briggs in 1995, important discoveries about the mechanisms of induction and maintenance of gLTP have been reported, including the finding that the response is uniquely dependent on serotonin for both the induction phase and maintenance phase. These new advances will be discussed in depth in this review. PMID- 15784302 TI - Neuromelanin in human dopamine neurons: comparison with peripheral melanins and relevance to Parkinson's disease. AB - Neuromelanin (NM) is a dark polymer pigment produced in specific populations of catecholaminergic neurons in the brain. It appears in greatest quantities in the human brain, in lesser amounts in some other non-human primates, but is absent from the brain in many lower species. Interest in this pigment has seen a resurgence in recent years because of a hypothesised link between neuromelanin and the especial vulnerability of neuromelanin-containing neurons to cell death in Parkinson's disease (PD). Little is known regarding the biology of neuromelanin. As neuromelanin appears to have characteristics in common with the better studied peripheral melanin pigments this review compares what is known about neuromelanin with melanins found in other body tissues. Unlike peripheral melanins, which are produced in specialised cells called melanocytes and may be transferred to other cell types, neuromelanin granules are believed to be stored in the cell in which they are produced. Neuromelanin granules display a unique, more heterogeneous appearance compared with peripheral melanins. Unlike melanin, neuromelanin is traditionally thought to result from a non-enzymatic synthesis pathway with no known pathway for neuromelanin catabolism. More recent data, however, is indicative of some regulation of neuromelanin synthesis and turnover. By analogy with peripheral melanins, neuromelanin may function in vivo to attenuate the effects of damaging stimuli. Among several possible mechanisms suggested, the ability of neuromelanin to interact with transition metals, especially iron, and to mediate intracellular oxidative mechanisms has received particular attention. Recent data from neuromelanin in the Parkinson's disease brain suggests that this proposed function may be compromised, thus rendering pigmented neurons vulnerable to oxidative damage in this disorder. PMID- 15784303 TI - The reticular nucleus revisited: intrinsic and network properties of a thalamic pacemaker. AB - The intrinsic and network properties of thalamic reticular (RE) neurons, which release the potent inhibitory neurotransmitter gamma-aminobutyric acid (GABA), endow them with oscillatory properties within the frequency range of sleep spindles (7-15 Hz), a hallmark brain rhythm that characterizes early sleep stages. The original hypothesis that RE neurons are pacemakers of spindles, based on absence of this oscillation in thalamocortical (TC) systems after disconnection from RE nucleus and presence of spindle rhythmicity in the deafferented RE nucleus, is supported by new experimental results in vivo, in vitro and in computo showing that interactions through chemical synapses as well as electrical coupling among inhibitory RE neurons lead to generation and synchronization of spindle sequences within the nucleus. Besides their pacemaking role in spindle generation, RE neurons are crucially implicated in the inhibition of TC neurons during cortically generated spike-wave (absence) seizures, which may explain the obliteration of signals from the external world and unconsciousness during these epileptic fits. PMID- 15784305 TI - Use of veterinary antiprotozoal products in the UK, 1998-2002. AB - The Veterinary Medicines Directorate, an agency of the Department for Environment, Food and Rural Affairs, collates and publishes reports detailing the quantities of veterinary antimicrobial products sold in the UK by the pharmaceutical companies. For the most part, these reports have focussed on antibacterials. This paper reports, for the first time, the sales of antiprotozoal products (as base active ingredients) in the UK over a five-year period, 1998-2002, to act as a baseline. Trends and patterns in sales of different groups of the antiprotozoal products are also evaluated. It is hoped that knowledge of sales of these products will fill the knowledge gap of the potential for overall development of antimicrobial resistance in the veterinary field and also reinforce the commitment of the UK Government to promote responsible use of antimicrobial products in the veterinary field. PMID- 15784304 TI - A review of systems and networks of the limbic forebrain/limbic midbrain. AB - Evolutionarily older brain systems, such as the limbic system, appear to serve fundamental aspects of emotional processing and provide relevant and motivational information for phylogenetically more recent brain systems to regulate complex behaviors. Overall, overt behavior is, in part, determined by the interactions of multiple learning and memory systems, some seemingly complementary and some actually competitive. An understanding of limbic system function in emotion and motivation requires that these subsystems be recognized and characterized as extended components of a distributed limbic network. Behavioral neuroscientists face the challenge of teasing apart the contributions of multiple overlapping neuronal systems in order to begin to elucidate the neural mechanisms of the limbic system and their contributions to behavior. One major consideration is to bring together conceptually the functions of individual components of the limbic forebrain and the related limbic midbrain systems. For example, in the rat the heterogeneous regions of the prefrontal cortex (e.g., prelimbic, anterior cingulate, subgenual cortices and orbito-frontal areas) make distinct contributions to emotional and motivational influences on behavior and each needs consideration in its own right. Major interacting structures of the limbic system include the prefrontal cortex, cingulate cortex, amygdaloid nuclear complex, limbic thalamus, hippocampal formation, nucleus accumbens (limbic striatum), anterior hypothalamus, ventral tegmental area and midbrain raphe nuclei; the latter comprising largely serotonergic components of the limbic midbrain system projecting to the forebrain. The posterior limbic midbrain complex comprising the stria medullaris, central gray and dorsal and ventral nuclei of Gudden are also key elements in the limbic midbrain. Some of these formations will be discussed in terms of the neurochemical connectivity between them. We put forward a systems approach in order to build a network model of the limbic forebrain/limbic midbrain system, and the interactions of its major components. In this regard, it is important to keep in mind that the limbic system is both an anatomical entity as well as a physiological concept. We have considered this issue in detail in the introduction to this review. The components of these systems have usually been considered as functional units or 'centers' rather than being components of a larger, interacting, and distributed functional system. In that context, we are oriented toward considerations of distributed neural systems themselves as functional entities in the brain. PMID- 15784306 TI - Susceptibility patterns of Streptococcus pneumoniae isolates in North America (2002-2003): contemporary in vitro activities of amoxicillin/clavulanate and 15 other antimicrobial agents. AB - A contemporary (2002-2003) national collection of 2100 strains of Streptococcus pneumoniae obtained from 30 sites in the nine United States (US) census regions were tested to determine the comparative antimicrobial properties of amoxicillin/clavulanate and 15 other antimicrobials. The rank order of antimicrobials with the lowest susceptibility rates was: penicillin (67.9%)or=41.1%), trimethoprim/sulphamethoxazole (38.9%), tetracyclines (22.2%) and clindamycin (10.0%). Geographical variation in the susceptibility patterns among US census zones was present with lowest penicillin and erythromycin susceptibility noted for West South Central and West North Central zones (or=+0.9%), sinus isolates (+2.7%), middle ear fluid isolates (+5.5%), penicillin-resistant strains (>or=+5.8%) and strains from patients <2 years of age (>or=+2.4%). Local and global surveillance studies of common respiratory pathogens such as S. pneumoniae remain instrumental to guide clinicians in appropriate empirical treatments and to emphasize the need for prudent antimicrobial use. PMID- 15784307 TI - Alterations in the GyrA and GyrB subunits of topoisomerase II and the ParC and ParE subunits of topoisomerase IV in ciprofloxacin-resistant clinical isolates of Pseudomonas aeruginosa. AB - The presence of fluoroquinolone resistance-associated alterations in topoisomerase II and IV were investigated for 103 nfxC-like type Pseudomonas aeruginosa isolates. The most nfxC-like type isolates (98.1%) possessed the substitution of Ile for Thr-83 in GyrA. A single alteration in GyrA (Thr-83- >Ile) was the most frequently detected and the next common alteration was two alterations with Thr-83-->Ile in GyrA and Ser-87-->Leu in ParC. A novel alteration at position Glin-106 of GyrA, which was suggested to be responsible for fluoroquinolone resistance, was identified. Our study revealed that the alterations in GyrB (Glu-468-->Asp) and in ParE (Asp-419-->Asn or Glu-459-->Asp) play a complementary role in the acquisition of resistance to fluoroquinolone. There was a correlation between the ciprofloxacin MIC and the number of resistance-associated alterations in GyrA, GyrB, ParC and ParE of P. aeruginosa isolates. PMID- 15784308 TI - Evaluation of antimicrobial activity of beta-lactam antibiotics using Etest against clinical isolates from 60 medical centres in Japan. AB - An antimicrobial resistance surveillance study was carried out in 60 medical centres across Japan. Resistance to piperacillin was 10.8% in clinical isolates of Escherichia coli, while 1.3% or fewer isolates were resistant to other beta lactams. Klebsiella spp. were more susceptible to imipenem, cefepime and cefpirome. Isolates of Enterobacter spp., Citrobacter spp., indole-positive Proteus and Serratia spp. were susceptible to imipenem, cefepime and cefpirome, while Acinetobacter spp. were most susceptible to cefoperazone/sulbactam, imipenem, ceftazidime (5.8% resistance) and cefepime (7.6%). Isolates of Pseudomonas aeruginosa were more susceptible to ceftazidime (12.3% resistance), cefoperazone/sulbactam (12.5%) and cefepime (12.6%) than to piperacillin (15.0%), cefpirome (22.6%) and imipenem (30.8%). The percentage of Japanese imipenem resistant P. aeruginosa clinical isolates was around 30%. PMID- 15784309 TI - Comparative activity of quinolones, macrolides and ketolides against Legionella species using in vitro broth dilution and intracellular susceptibility testing. AB - The comparative in vitro activity of quinolones (trovafloxacin, gemifloxacin, levofloxacin, ciprofloxacin, moxifloxacin and grepafloxacin), ketolides (ABT-773 and telithromycin) and macrolides (clarithromycin, azithromycin and erythromycin) were evaluated against Legionella pneumophila by broth dilution and an HL-60 intracellular model. The MIC90 of the quinolones, clarithromycin and ABT-773 were more than eight times lower than for erythromycin. Telithromycin, ABT-773 and azithromycin had significantly greater intracellular activity against L. pneumophila than erythromycin at 1xMIC and 8xMIC. The rank order of intracellular activity against L. pneumophila serogroup 1 was quinolones>ketolides>macrolides. Clinical trials to determine the clinical efficacy of ketolides for the treatment of Legionnaires' disease are warranted. PMID- 15784310 TI - Acquired sulphonamide resistance genes in faecal Escherichia coli from healthy children in Bolivia and Peru. AB - Antimicrobial resistance and sulphonamide resistance determinants were studied in 20 co-trimoxazole resistant Escherichia coli in faecal samples from healthy children in Bolivia and Peru. Methods used were disc diffusion susceptibility tests, PCR, sequence analysis and plasmid conjugation assays. All isolates but one were resistant to at least two different classes of antimicrobials; 19 isolates also carried at least one sul-gene. The most frequent gene was sul2 followed by sul1 and sul3, which was detected in one isolate. This is the first observation of sul3 on the American continent. In conclusion, the high prevalence of sul-genes in this material of faecal commensal E. coli isolates points to a potential role of the faecal flora in the emergence and spread of antimicrobial resistance. PMID- 15784311 TI - Identification of a novel antifungal nonapeptide generated by combinatorial approach. AB - It is becoming clear that antimicrobial peptides are important components of the innate defences of all species of life. They kill very rapidly, do not easily select resistant mutants and are synergistic with potentially toxic conventional therapeutic agents against microbes. This paper describes an attempt to expand a lead hexapeptide motif synthesized through combinatorial approach. A cationic peptide H-Arg-Trp-Trp-Arg-D-Trp-D-Phe-Ile-D-Phe-His-NH2 was found to be active with a therapeutic index of >17. I was proposed that the combination of peptide with known antifungal agents may identify synergistic combinations that would ideally reduce the dosage of conventional antifungals as well as their associated toxicity. Nine different pathogenic strains and species of Candida and two of Cryptococcus neoformans were employed in chequerboard method and in time kill assays to evaluate the synergistic effect of the lead peptide in combination with amphotericin B, 5-flucytosine, ketoconazole and fluconazole. We found synergistic interaction between the peptide and all four drugs against Cryptococcus isolates whilst both synergistic and additive combinations occurred when Candida isolates were used. PMID- 15784312 TI - In vitro susceptibility testing of Candida and Aspergillus spp. to voriconazole and other antifungal agents using Etest: results of a French multicentre study. AB - Minimum inhibitory concentrations (MICs) of the antifungal agent voriconazole were determined using the Etest and compared with those of amphotericin B, itraconazole and fluconazole using 1986 clinical isolates of Candida spp. Voriconazole MICs were also compared with those of amphotericin B and itraconazole using 391 clinical isolates of Aspergillus spp. Voriconazole was found to have more potent activity and lower MIC values than amphotericin B, itraconazole and fluconazole against C. albicans, C. tropicalis, C. parapsilosis and C. kefyr. Against C. glabrata and C. krusei, voriconazole was more active than either of the other two azole antifungals but had similar activity to amphotericin B. For species of Aspergillus, MIC values of voriconazole were lower than those of amphotericin B and itraconazole against A. fumigatus and A. flavus, and were similar to those of amphotericin B against A. niger. Against A. terreus, MIC values for voriconazole and itraconazole were similar. A. terreus is known to be resistant to amphotericin B, and this was reflected in higher MIC values compared with those of voriconazole and itraconazole. Voriconazole therefore compares very favourably with other antifungal agents against a large number of clinical isolates of Candida and Aspergillus spp. PMID- 15784313 TI - Blood culture Gram stain and clinical categorization based empirical antimicrobial therapy of bloodstream infection. AB - Early antimicrobial treatment has a great influence on the outcome of patients with blood stream infections (BSI). The study was designed to see if the simple practice of patient categorization (community acquired, nosocomial or infection in haematological unit) combined with Gram stain data could be used to guide empirical treatment of BSI in 1901 consecutive positive blood culture findings. There were considerable differences in the occurrence of common pathogens and their antimicrobial susceptibilities between patient categories especially for Gram-positive cocci. For example, second generation cephalosporins covered more than 70% cocci in clusters and over 80% of cocci in chains in community acquired infections whereas in hospital acquired infections the corresponding figures were only 47 and 44%. We conclude that Gram stain results of positive blood cultures along with the knowledge of where the infection was acquired, would allow early accurate targeting of antimicrobial therapy for BSI. PMID- 15784314 TI - Increased antibacterial activity of DW286, a novel fluoronaphthyridone antibiotic, against Staphylococcus aureus strains with defined mutations in DNA gyrase and topoisomerase IV. AB - To investigate the activity of DW286, a new fluoronaphthyridone, the quinolone resistance determining regions (QRDRs) of gyrA, gyrB, grlA and grlB genes in 64 Staphylococcus aureus clinical isolates were analyzed and the MICs of DW286 and comparator quinolones determined. Double and triple mutants in gyrA and grlA were resistant to ciprofloxacin, sparfloxacin, trovafloxacin and gemifloxacin but susceptible to DW286 (MIC 0.25-0.5 mg/l). The fourth alteration, Ser85Pro of GyrA was required to make a strain resistant to DW286 (MIC 4-32 mg/l). For a strain with the mutations at GyrA Ser84Leu and GrlA Ser80Phe, the MBC of DW286 was two fold higher than its corresponding MIC, in contrast to ciprofloxacin which was not bactericidal. PMID- 15784315 TI - 3,4,5,3',5'-Pentabromo-2-(2'-hydroxybenzoyl)pyrrole: a potential lead compound as anti-Gram-positive and anti-biofilm agent. AB - The activity against Gram-positive bacteria of 3,4,5,3',5'-pentabromo-2-(2' hydroxybenzoyl)pyrrole I, a synthetic anti-bacterial compound related to pyrrolomycins, was tested in vitro using seven reference bacterial strains and Staphylococcus epidermidis and Staphylococcus aureus preformed biofilms. Compound I was active against all strains tested, with minimum inhibitory concentration (MIC) values ranging from 0.002 to 0.097 mg/l and minimum bactericidal concentrations (MBCs) from 0.37 to 12.5 mg/l. Compound I was also active at low concentrations against preformed S. epidermidis and S. aureus biofilms. PMID- 15784317 TI - Salmonella enterica serovar Typhi with decreased susceptibility to ciprofloxacin- an emerging problem in Bangladesh. PMID- 15784316 TI - Tn7 distribution in Helicobacter pylori: a selective paradox. AB - The presence of class 1, 2 and 3 integrons in Helicobacter pylori, which in other species are usually related to antimicrobial resistance, were looked for in 40 epidemiologically-unrelated isolates. The presence of the class 2 integrase, which led to the discovery of Tn7, was found in 15 of 35 different clones identified by lspA-glmM polymorphism. MIC values for trimethoprim and streptomycin were not affected by the presence or absence of dfrA1 and aadA1 genes. The polyclonal spread of Tn7 showed the ability of H. pylori to harbour antimicrobial resistance mechanisms, usually located on plasmids, and also suggested that other important mechanisms of resistance may also be acquired, which would result in failure of chemotherapy. PMID- 15784318 TI - Antimicrobial susceptibility of Aeromonas spp., Vibrio spp. and Plesiomonas shigelloides isolated in the Philippines and Thailand. PMID- 15784319 TI - Emergence of CTX-M-type beta-lactam resistance in Shigella spp. in Hong Kong. PMID- 15784320 TI - Optimal co-amoxiclav ratios for inhibiting Escherichia coli strains with different susceptibilities to the compounds. PMID- 15784321 TI - Melting characteristics of fat present on the surface of industrial spray-dried dairy powders. AB - The melting characteristics of the fat present on the surface (surface free-fat) of two industrial spray-dried dairy powders (cream powder and whole milk powder) were investigated in comparison with those of other milk fat fractions present in the powder, such as free-fat from the interior of the powder particle (inner free fat) and encapsulated fat. The melting characteristics of the milk fat fractions were studied by fatty acid composition, melting profile and solid fat content profile. The results indicated that all milk fat fractions including surface free fat contained various triglycerides with melting points ranging from -40 to +40 degrees C. However, some fractionation was observed among the different milk fat fractions. The free-fat fractions (surface free-fat and inner free-fat) had a greater proportion of high-melting triglyceride species than the encapsulated fat. Furthermore, the high-melting triglyceride species present in the free-fat fractions were slightly accumulated at the surface of powder. This phenomenon was observed in both cream powder and whole milk powder and its effect on wetting time was established. This indicates that manipulation of the surface fat content during drying operation may hold the key to functionality improvement. PMID- 15784322 TI - A kinetic study of the formation of beta-cyclodextrin complexes with monomolecular films of fatty acids and glycerides spread at the air/water interface. AB - The kinetics of formation of inclusion complexes between beta-cyclodextrin and monolayers of one-, two- and three-chained lipid molecules, namely, oleic acid (OA), monoolein (MO), diolein (DO) and triolein (TO), was investigated at various pH using three independent dynamic methods. The formation and solubilization of soluble inclusion beta-CD/OA and beta-CD/MO complexes was detected by measuring the decrease of the surface area and surface pressure of the OA and MO monolayers in the presence of beta-CD within a wide range of concentrations. A third approach, describing the dilatational properties of the monolayers, influenced by the formation and solubilization of the complexes, was developed. Using the three above-mentioned independent methods, the rate constants of formation (k1) and dissociation (k2) of beta-CD/OA and beta-CD/MO, were determined. We observed that solubilization flux i s for OA monolayer increases with pH and at pH 11 reached a value, which is closed to the diffusion flux iD and the process thus becomes diffusion controlled. For MO monolayer no significant effects of pH was observed above pH 6. The surface pressure (Deltapi)--area per molecule (A) and surface potential (DeltaV)--area per molecule (A) isotherms and rheological properties of DO and TO monolayers were measured in the presence or absence of beta-CD. DO and TO form water-insoluble complexes with beta-CD, as visualized by AFM images. PMID- 15784323 TI - Structural properties and Raman spectroscopy of lipid Langmuir monolayers at the air-water interface. AB - Spectra of octadecylamine (ODA) Langmuir monolayers and egg phosphatidylcholine (PC)/ODA-mixed monolayers at the air-water interface have been acquired. The organization of the monolayers has been characterized by surface pressure-area isotherms. Application of polarized optical microscopy provides further insight in the domain structures and interactions of the film components. Surface enhanced Raman scattering (SERS) data indicate that enhancement in Raman spectra can be obtained by strong interaction between headgroups of the surfactants and silver particles in subphase. By mixing ODA with phospholipid molecules and spreading the mixture at the air-water interface, we acquired vibrational information of phospholipid molecules with surfactant-aided SERS effect. PMID- 15784324 TI - Surface modification of surface sol-gel derived titanium oxide films by self assembled monolayers (SAMs) and non-specific protein adsorption studies. AB - Biological events occurring at the implant-host interface, including protein adsorption are mainly influenced by surface properties of the implant. Titanium alloys, one of the most widely used implants, has shown good biocompatibility primarily through its surface oxide. In this study, a surface sol-gel process based on the surface reaction of metal alkoxides with a hydroxylated surface was used to prepare ultrathin titanium oxide (TiOx) coatings on silicon wafers. The oxide deposited on the surface was then modified by self-assembled monolayers (SAMs) of silanes with different functional groups. Interesting surface morphology trends and protein adhesion properties of the modified titanium oxide surfaces were observed as studied by non-specific protein binding of serum albumin. The surface properties were investigated systematically using water contact angle, ellipsometry, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM) measurements. Results showed that the surface sol-gel process predominantly formed homogeneous, but rough and porous titanium oxide layers. The protein adsorption was dependent primarily on the silane chemistry, packing of the alkyl chains (extent of van der Waals interaction), morphology (porosity and roughness), and wettability of the sol-gel oxide. Comparison was made with a thermally evaporated TiOx-Ti/Si-wafer substrate (control). This method further extends the functionalization of surface sol-gel derived TiOx layers for possible titanium alloy bioimplant surface modification. PMID- 15784325 TI - Langmuir monolayers of the long-chain alkyl derivatives of a nucleoside analogue and the formation of self-assembled nanoparticles. AB - The long-chain alkyl derivatives of a nucleoside analogue-acyclovir were prepared in the paper. One is stearyl-glycero-succinyl-acyclovir (SGSA) with a single 18 carbon length (C18) alkyl chain. Another is dioctadecyl-aspartate-succinyl acyclovir (DASA) with double C18 alkyl chains. They were prepared by the esterification of succinyl-acyclovir with the lipids, and sodium salts of them were also prepared. Guanine moieties and alkyl moieties bring the derivatives intermolecular hydrogen bonding and hydrophobic interaction in water separately. The forces are influenced by the number of alkyl chains and the charged state, and determine the solubility and the self-assembly behavior of the derivatives. The double alkyl-chain derivatives (DASA and DASA-Na) formed rigid Langmuir monolayers on air/water surface, while the single alkyl chain derivatives (SGSA and SGSA-Na) did not. However, cholesterol (Chol) could assist SGSA to form rigid monolayers through inserting into the alkyl chains of SGSA to mimic the second alkyl chain. SGSA self-aggregates in water were prepared by the injection method with tetrahydrofuran as solvent. Cuboid-like shape and nanoscale size demonstrated that SGSA self-aggregates were self-assembled nanoparticles. Shape, particle size, zeta potential and phase transition of the nanoparticles were characterized. And they showed an average size of 83.2 nm, a negative surface charge of -31.3-mV zeta potential and a gel-liquid crystalline phase transition of 50.38 degrees C. The formation mechanism of self-assembled nanoparticles was analyzed. Hydrophobic interaction of alkyl chains improves SGSA molecules to form bilayers, and then cuboid-like nanoparticles were obtained by layer-by-layer aggregation based on inter-bilayers hydrogen bonding. However, the charged guanine moieties make SGSA-Na lose the function of hydrogen bonding so that SGSA Na only forms vesicles in water based on hydrophobic interaction. Strong hydrophobicity and wide-open rigid double alkyl chains of DASA and DASA-Na restrict self-assembly in water media, and no homogeneous suspensions were obtained. Therefore, the molecular self-assembly behavior of the long-chain alkyl derivatives of nucleoside analogues on water surface or in water media is determined by the number of alkyl chains and the charged state. PMID- 15784326 TI - Colloidal stability of aqueous polymeric dispersions: effect of pH and salt concentration. AB - Aqueous film coatings often contain some electrolytes, organic acids, and pigments to give functions of sustained release, time-controlled release, or protection against light. Additions of some electrolytes or organic acids into latex dispersion for an aqueous film coating affect its colloidal stability. We characterized the aqueous polymeric latexes used in the pharmaceutical industry by measuring zeta potential and particle size, and evaluated this colloidal stability using DLVO theory. Three polymethacrylate-based aqueous polymeric latexes, Eudragit L30D-55, Eudragit RS30D and Eudragit NE30D, having anionic, cationic, and neutral polymer, respectively, were used in this study. The Hamaker constant of the polymethacrylate-based latex was determined to be 6.35 x 10(-21) J, and the total potential energy of the latex dispersion was calculated. The total potential energy of interaction between pairs of latex particles changes by altering the salt concentration and pH. The experimental results of stability in the anionic and the cationic latex dispersions can be explained by the total interaction energies. However, the stabilization of the neutral latex did not match the calculated result. The steric interaction produced by the surfactant likely resulted in the stable dispersion of this latex. PMID- 15784327 TI - Interaction of gemini surfactants butane-1,4-diyl-bis(alkyldimethylammonium bromide) with DNA. AB - The size and structure of aggregates formed by interaction of DNA with homologous series of cationic gemini surfactants butane-1,4-diyl-bis(alkyldimethylammonium bromide) (CnGS, n=10-16 is the number of alkyl carbons) were investigated using UV-vis turbidity, dynamic light scattering and small-angle synchrotron X-ray (SAX) diffraction. The detailed analysis of turbidity in the range of lambda=450 600 nm indicates an anomaly in the growth of CnGS+DNA aggregates with increasing concentration of CnGS, possibly involving changes of structure and size of aggregates. Using dynamic light scattering, changes of the effective diameter of CnGS+DNA (n=12 and 16) aggregates formed in the CnGS concentration range 0.002 0.140 mmol/l were observed. SAX diffractograms show the presence of long-range organization of CnGS+DNA (n=12, 13, 14 and 16) aggregates due to DNA interaction with CnGS above the critical micellar concentration. The CnGS+DNA (n=12, 13 and 14) aggregates at 25 degrees C are packed in a lattice of two-dimensional hexagonal symmetry. With increasing C14GS:DNA molar ratio the changes of the lattice parameter in the range of 4.80-5.27 nm are observed at 25 degrees C. The aggregates undergo structural changes induced by temperature in the range 60-95 degrees C, which are accompanied by changes of the diffraction patterns, namely in the region of reciprocal spacing s=0.15-0.30 nm(-1). PMID- 15784328 TI - Modulation of lysozyme charge influences interaction with phospholipid vesicles. AB - Lysozyme is a globular protein which is known to bind to negatively charged phospholipid vesicles. In order to study the relationship between charge state of the protein and its interaction with negatively charged phospholipid membranes chemical modifications of the proteins were carried out. Succinylation and carbodiimide modification was used to shift the isoelectric point of lysozyme to lower and higher pH values, respectively. The binding of the modified lysozyme to phospholipid vesicles prepared from phosphatidic acid (PA) was determined using microelectrophoresis and ultracentrifugation. At acidic pH of the solution all lysozyme species reduced the surface charges of PA vesicles. Succinylated lysozyme (succ lysozyme) reduced the electrophoretic mobility (EPM) to nearly zero, whereas native lysozyme and carboxylated lysozyme (carbo lysozyme) changed the surface charge to positive values. At neutral pH, the reduction of surface charges was less for carbo lysozyme and unmodified lysozyme. Succ lysozyme did not change the EPM. Unmodified and carbo lysozyme decreased the magnitude of EPM, but the whole complex was still negatively charged. The bound fraction of all modified lysozyme to PA vesicles at high lysozyme/PA ratios was nearly constant at acidic pH. At low lysozyme/PA ratios the extent of bound lysozyme is changed in the order carbo>unmodified>succ lysozyme. Increasing the pH, the extent of bound lysozyme to PA large unilamellar vesicles (LUV) is reduced, at pH 9.0 only 35% of carbo lysozyme, 23% of unmodified lysozyme is bound, whereas succ lysozyme does not bind at pH 7.4 and 9.0. At low pH, addition of all lysozyme species resulted in a massive aggregation of PA liposomes, at neutral pH aggregation occurs at much higher lysozyme/PA ratios. Lysozyme binding to PA vesicles is accompanied by the penetration of lysozyme into the phospholipid membrane as measured by monolayer techniques. The penetration of lysozyme into the monolayer was modulated by pH and ionic strengths. The interaction of lysozyme with negatively charged vesicles leads to a decrease of the phospholipid vesicle surface hydration as measured by the shift of the maximum of the fluorescence signal of a headgroup labeled phospholipid. The binding of bis-ANS as an additional indicator for the change of surface hydrophobicity is increased at low pH after addition of lysozyme to the vesicles. More hydrophobic patches of the lysozyme-PA complex are exposed at low pH. At low pH the binding process of lysozyme to PA vesicles is followed by an extensive intermixing of phospholipids between the aggregated vesicles, accompanied by a massive leakage of the vesicle aqueous content. The extent of lysozyme interaction with PA LUV at neutral and acidic pH is in the order carbo lysozyme>lysozyme>succ lysozyme. PMID- 15784329 TI - Effect of pressure on the Prodan fluorescence in bilayer membranes of phospholipids with varying acyl chain lengths. AB - The fluorescence spectra of 6-propionyl-2-(dimethylamino)naphthalene (Prodan) were observed as a function of pressure for the bilayer membrane systems of dilauroylphosphatidylcholine (DLPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), and distearoylphosphatidylcholine (DSPC). The wavelength of the emission maximum, lambdamax, was found to be 480, 430, and 500 nm for the liquid crystalline (Lalpha), ripple gel (P'beta), and pressure induced interdigitated gel (LbetaI) phase, respectively. Since the lambdamax reflects the solvent property around the probe molecules, we could speculate on the location of the Prodan molecules in the bilayer membranes; in the Lalpha phase of the lipid bilayer, the Prodan molecules distribute around the phosphate of the lipids (i.e. the polar region). The Lalpha/P'beta phase transition caused the Prodan molecules to move into the less polar region near the glycerol backbone. The fluorescence intensity of the Prodan in the P'beta phase was dependent on the chain length of the lipids and on pressure; the shorter the chain length of the lipid, the stronger the fluorescence intensity of the Prodan. Moreover, for the DLPC bilayer membrane system, the fluorescence intensity at 430 nm increased with increasing pressure, indicating that the partition of Prodan into the DLPC bilayer membrane is promoted by applying pressure. In the case of the DPPC and DSPC bilayers, as the pressure increased further, the pressure induced interdigitation caused the Prodan molecules to squeeze out of the glycerol backbone region and to move the hydrophilic region near the bilayer surface. The ratio of fluorescence intensity at 480 nm to that at 430 nm, F480/F430, showed a sharp change at the phase-transition pressure. In the case of the DPPC and DSPC bilayers, the values of F480/F430 showed an abrupt increase above a certain pressure higher than the Lalpha/P'beta transition pressure, which corresponds to the interdigitation from the P'beta to the LbetaI phase. The plot of F480/F430 versus pressure is available for recognition of the bilayer phase transitions, especially the bilayer interdigitation. PMID- 15784330 TI - Prediction of kinetics of doxorubicin release from sulfopropyl dextran ion exchange microspheres using artificial neural networks. AB - The purpose of this work was to develop artificial neural networks (ANN) models to predict in vitro release kinetics of doxorubicin (Dox) delivered by sulfopropyl dextran ion-exchange microspheres. Four ANN models for responses at different time points were developed to describe the release profiles of Dox. Model selection was performed using the Akaike information criterion (AIC). Sixteen data sets were used to train the ANN models and two data sets for the validation. Good correlations were obtained between the observed and predicted release profiles for the two randomly selected validation data sets. The difference factor (f1) and similarity factor (f2) between the ANN predicted and the observed release profiles indicated good performance of the ANN models. The established models were then applied to predict release kinetics of Dox from the microspheres of various initial loadings in media of different ionic strengths and NaCl/CaCl2 ratios. The results suggested that ANN offered a flexible and effective approach to predicting the kinetics of Dox release from the ion exchange microspheres. PMID- 15784331 TI - Pegylated nanoparticles based on poly(methyl vinyl ether-co-maleic anhydride): preparation and evaluation of their bioadhesive properties. AB - Pegylated nanoparticles based on poly(methyl vinyl ether-co-maleic anhydride) (PVM/MA) were prepared by simple solvent displacement method, in the absence of catalysts or specific chemical conditions. Pegylation efficiency increased with the increasing of molecular weight and bulk concentration of poly(ethylene glycols) (PEGs) investigated. In fact, the use of PEG with molecular weight less than 1000 Da did not lead to its attachment. 1H NMR spectroscopy was performed in order to estimate the conformation state of PEG-chains and to predict the nanoparticle structure. Pegylation with PEG 2000 gave surface modified nanoparticles ("brush" conformation), while the chains of PEG 1000 were distributed either in the core or physically adsorbed on the nanoparticle surface. The capacity of nanoparticles to adsorb mucin at pH 7.4 was significantly higher for PEG 1000-NP than for PEG 2000-NP. The "brush" layer seemed to decrease the interaction between PEG 2000-NP and mucin, which facilitated their penetration through the mucus gel. As a consequence, PEG 2000 NP displayed higher capacity to develop adhesive interactions with rat intestinal mucosa in vivo. Independent on the weaker bioadhesive potential of PEG 1000-NP, both types of pegylated nanoparticles demonstrated very high affinity to the intestinal mucosa rather than to the stomach wall, which could be established for drug targeting to the small intestine. PMID- 15784332 TI - Synthesis and pharmacological profile of non-peptide vasopressin antagonists. AB - Recently we presented a series of 6-ethyl and 6-benzylthieno[2,3-b][1,4]thiazine derivatives with relaxing effects on vascular smooth muscle and terminal ileum. In this report the synthesis of further thieno[2,3-b][1,4]thiazine derivatives and related compounds with a thieno[2,3-b][1,4]thiazepine or thieno[3,2 b][1,4]thiazine ring system is described. The pharmacological effect of the agents was tested in isolated smooth (terminal ileum, pulmonary artery, aortic rings, myometrial strips) and heart (papillary muscle, spontaneously beating right atrium) muscle preparations of the guinea pig. Contractions were measured isometrically, and smooth muscle preparations were either precontracted with high K+ (60 or 90 mM KCl containing nutrient solution) or with agonists, while papillary muscles were electrically stimulated (1 Hz). The vasopressin antagonistic activity of the test compounds was tested in isolated papillary muscles in which the V1A-receptor subtype is located. The biphasic response to vasopressin was antagonized, dependent on the chemical structure of the test compound. Thieno[3,2-b][1,4]thiazines were more potent than thieno[2,3 b][1,4]thiazine and thieno[2,3-b][1,4]thiazepine compounds. In addition, substitution of a methyl substituted terminal benzyl ring instead of a phenyl- or dichlorobenzoyl moiety attenuated the vasopressin antagonistic effect. PMID- 15784333 TI - Bioreversible quaternary N-acyloxymethyl derivatives of the tertiary amines bupivacaine and lidocaine--synthesis, aqueous solubility and stability in buffer, human plasma and simulated intestinal fluid. AB - Design of water-soluble prodrugs may constitute a means to improve the oral bioavailability of drugs suffering from dissolution rate-limited absorption. The model drug bupivacaine containing a tertiary amine function has been converted into bioreversible quaternary N-acyloxymethyl derivatives. The pH-independent solubility of the N-butanoyloxymethyl derivate exceeded 1000 mg ml-1 corresponding approximately to a 10,000-fold increase in water solubility compared to that of bupivacaine base. The kinetics of hydrolysis of the prodrugs was studied in the pH range 0.1-9.8 (37 degrees C). Decomposition was found to follow first-order kinetics and U-shaped pH-rate profiles were constructed. The observed differences between the hydrolytic lability of the derivatives might most likely be ascribed to steric effects. In most cases, the prodrugs were quantitatively converted into bupivacaine. However, for the hydrolysis of the N butanoyloxymethyl derivative at neutral to slightly alkaline pH parallel formation of bupivacaine (approximately 80%) and an unknown compound X (approximately 20%) was observed. LC-MS analysis of the latter compound suggests that an aromatic imide structure has been formed from an intramolecular acyl transfer reaction involving a nucleophilic attack of the amide nitrogen atom on the ester carbonyl carbon atom. Whereas the derivatives were poor substrates for plasma enzymes; they were hydrolyzed rapidly to parent bupivacaine in the presence of pancreatic enzymes (simulated intestinal fluid) at 37 degrees C. The data indicate that such prodrugs possess sufficient stability in the acidic environment of the stomach to reach the small intestine in intact form where they can be cleaved efficiently by action of pancreatic enzymes prior to drug absorption. Thus, the N-acyloxymethyl approach might be of potential utility to enhance oral bioavailability of tertiary amines exhibiting pKa values below approximately 6 and intrinsic solubilities in the low microM range. PMID- 15784334 TI - Amphiphilic amino acid copolymers as stabilizers for the preparation of nanocrystal dispersion. AB - The recent advance of particle size engineering in nanometer ranges has widened the formulation opportunities of relatively water-insoluble drugs. However, the 'nanoformulation' suffers from a lack of systematic understanding about the requirements of polymeric stabilizers. Furthermore, the polymers that can be used for the preparation of nanocrystals are so limited that finding a proper stabilizer for a given formulation is often difficult. In this study, amino acid copolymers whose properties can systematically be tailored are developed, and their morphological and compositional effects are investigated. Copolymers containing lysine (K) as their hydrophilic segments, and phenylalanine (F) or leucine (L) as their hydrophobic segments successfully produce stable nanocrystals (200-300 nm) in water, while copolymers of K and alanine (A) could not generate nanosized particles. Not the morphology but the hydrophobicity of copolymers seems to be a critical parameter in the preparation of drug nanocrystals by wet comminution. The effective stabilization performance of copolymers requires the hydrophobic moiety content to be higher than 15 mol%. Comminution for only 5 min is long enough for nanocrystal preparation, and the crystallinity of drug is found intact after the processing. PMID- 15784335 TI - Multivariate modeling of cytochrome P450 3A4 inhibition. AB - In the early phases of current pharmaceutical research projects, huge numbers of compounds are tested on their biological activity with respect to a certain target by experimental or virtual screening campaigns. To reduce the attrition rate in later stages of a project, other relevant properties such as physicochemical and ADMET (absorption, distribution, metabolism, excretion, toxicity) properties should be assessed as early as possible in lead discovery and optimization. The present study describes the development of in silico models to predict the inhibition of human cytochrome P450 3A4 (CYP3A4) from calculated molecular descriptors. The models were trained and validated using a set of 967 structural diverse drug-like research compounds with an experimentally determined CYP3A4 inhibition potency (IC50 value) which was carefully split into a training and a test set. For classification models, the data sets were further subdivided into strong, medium, and weak inhibitors. Different descriptor sets were used to cover various aspects of molecular properties, including properties derived from the 2D structure, the interaction of the molecule with its environment, and properties derived from quantum-mechanical calculations. The descriptors were related to the CYP3A4 inhibition potency by multivariate data analysis methods such as partial least-squares projection to latent structures (PLS), PLS discriminant analysis (PLS-DA), and soft independent class modeling (SIMCA). The squared correlation between experimental and predicted IC50 values of the previously unseen test set compounds was Qext2=0.6 for the best PLS models, corresponding to a root mean squared error (RMSE) of RMSE=0.45 (logarithm of IC50). The best PLS-DA models were able to correctly classify more than 60% of the test set compounds, whereas almost no strong inhibitors were wrongly classified as weak inhibitors and vice versa. Furthermore, relevant molecular properties were identified which are closely related to the CYP3A4 inhibition potency of a compound. The results presented here are very encouraging since our models could, for instance, serve to flag problematic compounds or to guide further synthesis efforts. PMID- 15784336 TI - Mixed-effects modelling of the interspecies pharmacokinetic scaling of pegylated human erythropoietin. AB - The aim of this study was to develop a population pharmacokinetic model for interspecies allometric scaling of pegylated r-HuEPO (PEG-EPO) pharmacokinetics to man. A total of 927 serum concentrations from 193 rats, 6 rabbits, 34 monkeys, and 9 dogs obtained after a single dose of PEG-EPO, administered by the i.v. (dose range: 12.5-550 microg/kg) and s.c. (dose range: 12.5-500 microg/kg) routes, were pooled in this analysis. An open two-compartment model with first order absorption and lag time (Tlag) and linear elimination from the central compartment was fitted to the data using the NONMEM V software. Body weight (WT) was used as a scaling factor and the effect of brain weight (BW), sex, and pregnancy status on the pharmacokinetic parameters was investigated. The final model was evaluated by means of a non-parametric bootstrap analysis and used to predict the PEG-EPO pharmacokinetic parameters in healthy male subjects. The systemic clearance (CL) in males was estimated to be 4.08WT1.030xBW-0.345 ml/h. In females, the CL was 90.7% of the CL in males. The volumes of the central (Vc) and the peripheral (Vp) compartment were characterized as 57.8WT0.959 ml, and 48.1WT1.150 ml, respectively. Intercompartmental flow was estimated at 2.32WT0.930 ml/h. Absorption rate constant (Ka) was estimated at 0.0538WT-0.149. The absolute s.c. bioavailability F was calculated at 52.5, 80.2, and 49.4% in rat, monkey, and dog, respectively. The interindividual variability in the population pharmacokinetic parameters was fairly low (<35%). Non-parametric bootstrap confirmed the accuracy of the NONMEM estimates. The mean model predicted pharmacokinetic parameters in healthy male subjects of 70 kg were estimated at: CL: 26.2 ml/h; Vc: 3.6l; Q: 286 l/h; Vp: 6.9l, and Ka: 0.031 h-1. The population pharmacokinetic model developed was appropriate to describe the time course of PEG-EPO serum concentrations and their variability in different species. The model predicted pharmacokinetics of PEG-EPO in humans suggest a less frequent dosing regimen relative to erythropoietin and darbepoetin, potentially leading to a simplification of anemia management. PMID- 15784337 TI - Effect of self-microemulsifying drug delivery systems containing Labrasol on tight junctions in Caco-2 cells. AB - The aim of this study was to investigate the effect of two novel self microemulsifying drug delivery systems (SMEDDS) containing Labrasol with different dilutions on tight junctions. Changes in barrier properties of Caco-2 cell monolayers, including transepithelial electrical resistance (TEER) and permeability to the paracellular marker, i.e., mannitol, were assessed in response to dilutions and surfactants contents within formulations. The cytotoxicity of SMEDDS and the effect of surfactants on Caco-2 cells were evaluated by the MTT. Changes in subcellular localization of the tight junction proteins, ZO-1 and F-actin, were examined by confocal laser scanning microscopy. Results demonstrated that negatively charged SMEDDS with different dilutions had no effect on the TEER, but significantly increased the permeability of mannitol. In contrast, the positively charged formulation showed a dilution-dependent reduction in TEER. A corresponding increase in mannitol permeability of up to 29.4-fold to 64.7-fold greater than the control was also observed across the monolayer. Labrasol with the concentration of 0.1 and 1% was shown to increase the permeability of mannitol by 4.6-fold and 33.8-fold, respectively. The mechanism of opening of tight junctions was found to involve F-actin-related changes and redistribution of ZO-1. PMID- 15784338 TI - Preparation and evaluation of release characteristics of 3TabGum, a novel chewing device. AB - A new chewing gum device in the form of a three layers tablet (3TabGum) has been developed. The new drug delivery system is obtained, at room temperature, by direct compression using conventional pharmaceutical equipment. Basically, the resulting chewing gum tablets comprise a gum core combined with two protective antiadherent external layers, which prevent gum adhesion to the punches of the tableting machine. Drug release from a dosage form is the critical step in drug absorption and bioavailability, thus an experimental protocol has been designed to evaluate the efficiency of this kind of therapeutic system by verifying its capability to release the drug dose and by assessing the delivery rate. Simple diffusion into the medium causes the release of only a small percentage of the drug contained in the medicated chewing gum, while the delivery of the major part of the dose occurs during mastication. The results obtained in this study suggest that water soluble drugs are freely and easily released by chewing gums, while for actives characterized by reduced water solubility the release rate depends on the chewing time although all the drugs tested are completely released after a reasonable mastication time. PMID- 15784339 TI - Effects of miltefosine on membrane permeability and accumulation of [99mTc] hexakis-2-methoxyisobutyl isonitrile, 2-[18F]fluoro-2-deoxy-D-glucose, daunorubucin and rhodamine123 in multidrug-resistant and sensitive cells. AB - Miltefosine is a phospholipid analog that exhibits antineoplastic activity against breast cancer metastases, but its mechanism of action remains uncertain. The aim of this study was to investigate the transport mechanism for the removal of miltefosine and [99mTc]-hexakis-2-methoxyisobutyl isonitrile (99mTc-MIBI) from multidrug-resistant cells. The P-glycoprotein pump function, cell viability, and 99mTc-MIBI and 2-[18F]fluoro-2-deoxy-D-glucose (18FDG) uptakes were measured in NIH 3T3 (3T3) and NIH 3T3MDR1 G185 (3T3MDR1) mouse fibroblasts and human lymphoid B JY cells. Miltefosine treatment increased the permeability and fluidity of these tumor cells in a concentration-dependent manner. The multidrug-sensitive cells were 3-4 times more sensitive to miltefosine than the multidrug-resistant ones. The extent of 99mTc-MIBI accumulation in the P-glycoprotein-expressing cells increased in the presence of miltefosine, whereas the rhodamine123 and daunorubicin uptakes of the cells did not change significantly. In the 3T3MDR1 cells verapamil reinstated the rhodamine123 and daunorubicin accumulation, but not the 99mTc-MIBI uptake. Cyclosporin A reinstated the uptakes of 99mTc-MIBI, daunorubicin and rhodamine123 by the 3T3MDR1 cells. In a concentration-dependent manner miltefosine decreased the extents of 99mTc-MIBI, rhodamine123, daunorubicin and 18FDG accumulation in the JY and 3T3 cells. Our findings indicate a common transport mechanism for 99mTc-MIBI and miltefosine, which is distinct from that for rhodamine123 and daunorubicin in MDR cells. PMID- 15784340 TI - The role of physicochemical properties of entacapone and tolcapone on their efficacy during local intrastriatal administration. AB - Aqueous solubility, apparent partition coefficient (logPapp) and catechol-O methyltransferase (COMT, EC 2.1.1.6) inhibiting potency of entacapone and tolcapone were compared in vitro. Both drugs (at 10 and 100 microM) were also delivered directly into rat striatum via a microdialysis probe. Extracellular 3,4 dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) concentrations were measured to evaluate the inhibition of striatal COMT in vivo. Although entacapone had 15-fold better aqueous solubility than tolcapone at pH 7.4, also tolcapone had sufficient aqueous solubility to remain in solution at 100 microM. The logPapp of tolcapone was higher than that reported for entacapone in the pH range from 5.0 to 7.4. Entacapone and tolcapone inhibited equally rat striatal COMT in vitro with Ki values of 1.86 and 2.50 nM, respectively. Both drugs had similar outflow from the microdialysis probe in vitro. Perfusion of 100 microM entacapone increased significantly extracellular DOPAC levels compared to the control group. Both entacapone and tolcapone (at 10 and 100 microM) decreased significantly HVA levels, but entacapone was significantly more effective than tolcapone at 100 microM. In conclusion, entacapone and tolcapone are equally potent COMT inhibitors against rat striatal COMT in vitro. After local intrastriatal administration, entacapone appeared to inhibit COMT faster and more effectively than the more lipophilic tolcapone. Thus, intrastriatal administration led to opposite results compared to those reported in the brain after systemic administration. The present results also suggest that the local distribution of entacapone and tolcapone differ when the drugs are delivered directly into the brain. PMID- 15784341 TI - Transport of the investigational anti-cancer drug 5,6-dimethylxanthenone-4-acetic acid and its acyl glucuronide by human intestinal Caco-2 cells. AB - 5,6-Dimethylxanthenone-4-acetic acid (DMXAA), a potent cytokine inducer, exhibited marked antitumor activity when given as multiple oral doses in mice. The aim of this study was to examine the transport of DMXAA and its acyl glucuronide (DMXAA-G) using the human Caco-2 cells. DMXAA was minimally metabolized by Caco-2 cells and both DMXAA and DMXAA-G were taken up to a minor extent by the cells. The permeability coefficient (Papp) values of DMXAA over 10 500 microM were 4x10(-5) cm/s to 4.3x10(-5) cm/s for both apical (AP) to basolateral (BL) and BL-AP transport, while the Papp values for the BL to AP flux of DMXAA-G were significantly greater than those for the AP to BL flux, with Rnet values of 4.5-17.6 over 50-200 microM. The BL to AP active efflux of DMXAA-G followed Michaelis-Menten kinetics, with a Km of 83.5+/-5.5 microM, and Vmax of 0.022+/-0.001 nmol/min. The flux of DMXAA-G was energy and Na+-dependent and MK 571 significantly (P<0.05) inhibited its BL to AP flux, with an estimated Ki of 130 microM. These data indicate that the transport of DMXAA across Caco-2 monolayers was through a passive process, whereas the transport of DMXAA-G was mediated by MRP1/2. PMID- 15784342 TI - Enhancement of polymethacrylate-mediated gene delivery by Penetratin. AB - Polymethacrylates are vinyl-based polymers that are used for DNA transfection. Cationic polymethacrylates efficiently condense DNA by forming inter polyelectrolyte complexes. Their use for DNA transfection is, however, limited due to their low ability to interact with membranes. In order to increase their transfection efficiency, we combined polymethacrylates with Penetratin, a 16 residue water-soluble peptide that internalises into cells through membrane translocation. DNA condensation was assessed using physicochemical methods, while transfection efficiency and cellular internalisation were studied using Cos-1 cells. Agarose gel electrophoresis retardation, ethidium bromide exclusion tests and dynamic light scattering measurements showed that the stability of the polymethacrylate-DNA complexes is not affected by addition of Penetratin. Transfection efficiency of polymethacrylate-DNA complexes into Cos-1 cells increased by addition of Penetratin and was higher than that of polyethylenimine (PEI)-DNA complexes and comparable to Lipofectamine. Confocal microscopy and flow cytometry indicated that Penetratin mainly enhances endolysosomal escape polymethacrylate-DNA complexes and increases their cellular uptake. Since the cellular toxicity of polymethacrylate-DNA-Penetratin complexes remains low, especially compared to PEI, this transfection system opens new perspectives for gene therapy. PMID- 15784343 TI - Solubilization of poorly soluble lichen metabolites for biological testing on cell lines. AB - The depside atranorin and depsidone fumarprotocetraric acid, isolated from the lichens Stereocaulon alpinum and Cetraria islandica, respectively, were chosen as prototypes for poorly soluble natural compounds in an effort to facilitate testing in pharmacological models. Solubilizing agents previously identified as being non-toxic towards a malignant leukemic (K-562) cell line and suitable for testing of anti-proliferative activity of the dibenzofuran lichen metabolite (+) usnic acid were used in solubilization studies of the depside and depsidone. Cyclodextrin derivatives were found to be most suitable for solubilizing the lichen compounds, the greatest rise in solubility being witnessed for fumarprotocetraric acid, increasing almost 300-fold from 0.03 mg/ml in water to 8.98 mg/ml in 10% 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD). Subsequently, the lichen compounds, including (+)-usnic acid, were solubilized in 10% HPbetaCD and tested for effects on three malignant human cell lines; T-47D (breast), Panc-1 (pancreas) and PC-3 (prostate) in a standard proliferation assay. Atranorin and fumarprotocetraric acid did not exhibit anti-proliferative effects but usnic acid was active against all test cell lines with EC50 values of 4.3-8.2 microg/ml. The non-toxic solubilizing agents used in this study could prove useful for pharmacological testing of other poorly soluble natural products. PMID- 15784344 TI - Microcalorimetric studies of the effects on the interactions of human recombinant interferon-alpha2a. AB - The applicability of the physical stability of protein solution monitored by isothermal titration calorimetry (ITC) was evaluated. The second virial coefficient, b2, derived from the dilution enthalpies of protein solution measured by ITC under various experimental conditions was studied. The protein applied in this work is human recombinant interferon-alpha2a (hrIFN-alpha2a), which is a commercial drug applied for the treatment of virus-infected diseases. The results obtained were used to predict the possibility of hrIFN-alpha2a aggregation, and the prediction can be further confirmed by size-exclusion chromatography (SEC). Various factors affecting the stability of protein solution were investigated, for example, temperature, salts, surfactants, and mechanical stress. Specifically, the results show that the dilution enthalpy of hrIFN alpha2a increased with increasing temperature and NaCl concentration, while b2 decreased, indicating that the attraction between hrIFN-alpha2a molecules was enhanced under these conditions. On studying the effect of mechanical stress, the data obtained reveals that the introduction of centrifugal or vortex force strengthened the attractive forces between hrIFN-alpha2a molecules. These implications were supported by SEC data, demonstrating that the amount of aggregated hrIFN-alpha2a was increased. As a consequence, the methodologies presented in this investigation offer a possibility of monitoring the physical stability of protein solution at various stages of recovery, purification as well as the development of appropriate drug storage formulations. PMID- 15784345 TI - Physicochemical evaluation of PVP-thiazide diuretic interactions in co-spray dried composites--analysis of glass transition composition relationships. AB - The aim of this study was to evaluate the possible interactions in the solid state between the thiazide diuretics: bendroflumethiazide (BFMT), hydroflumethiazide (HFMT) and hydrochlorothiazide (HCT) and polyvinylpyrrolidone (PVP) following processing. The glass transition temperatures (T(g)s) of a range of binary co-spray-dried PVP-thiazide composites were determined and compared to the predictions of the Gordon-Taylor, Fox, Couchman-Karasz, Kwei and Schneider equations. The solid composites of the thiazide diuretics and PVP were prepared by a spray drying technique. Properties of composites were determined with the use of helium pycnometry and FTIR spectroscopy. For many systems studied the experimentally detected T(g)s exhibited large positive deviations when compared with the values predicted by the Gordon-Taylor, Fox and Couchman-Karasz equations. The data was better fitted by the Schneider equation consistent with a drug-polymer interaction. FTIR analysis revealed that strong hydrogen bonding between the sulphonamide groups of the thiazide diuretics and the PVP molecule was responsible for the increase in the T(g)s. Additionally, in the case of BFMT PVP composites, an interaction between the phenyl group and polymer ring was apparent. Glass transition-composition behaviour for amorphous drug-PVP composites deviated from the predictions of the commonly used Gordon-Taylor equation. Deviations were consistent with interactions between the components in the amorphous mixtures. The Schneider equation may be successfully applied to fit the Tg-composition profiles obtained, where other models fail to give good predictions. PMID- 15784346 TI - Transfer of morphine along the olfactory pathway to the central nervous system after nasal administration to rodents. AB - The aim of this study was to investigate whether morphine can be transferred along the olfactory pathway to the CNS, thereby circumventing the blood-brain barrier, after nasal administration to rodents. Radiolabelled and unlabelled morphine were administered via the right nostril to mice and rats. Olfactory bulbs, brain tissue and blood samples were collected. Morphine-derived radioactivity was measured using liquid scintillation (LS) and the concentrations of morphine and its metabolite morphine-3-glucuronide (M3G) were also assessed with high-performance liquid chromatography. The location of morphine-derived radioactivity in the rat brain was visualised by autoradiography. Overall, the levels of morphine in the right olfactory bulbs (ROBs) significantly exceeded those in the left olfactory bulbs (LOBs) and brain tissue samples 15, 60 and 240 min after right-sided nasal administration. Fifteen minutes after intravenous administration, there were no significant differences between olfactory bulbs and the other brain areas. Five minutes after nasal administration, autoradiography revealed radioactivity surrounding the ROB and reaching one of the ventricles in the brain. After 60 min, radioactivity had reached the peripheral parts of the ROB. All the techniques used in this study demonstrate that morphine was transferred along the olfactory pathway to the CNS after nasal administration to rodents. PMID- 15784347 TI - The potential impact of percutaneous pulmonary valve stent implantation on right ventricular outflow tract re-intervention. AB - OBJECTIVE: To assess the impact of a percutaneous technique for pulmonary valve implantation on the conventional surgical valve/conduit approach to right ventricular outflow tract re-intervention. METHODS: We have retrospectively reviewed our results following surgical or percutaneous re-intervention to the right ventricular outflow tract in both paediatric and adult groups. Between November 1998 and March 2004, 94 patients underwent surgical re-intervention to the right ventricular outflow tract. Percutaneous pulmonary valve implantation was introduced in October 2002 and 35 procedures were performed to March 2004. The median age was 26 years (6-65 years) in the surgical group and 16 years (9-39 years) in the percutaneous group. Tetralogy of Fallot was the commonest original diagnosis (64.9 and 62.9%, respectively). The predominant indication for re intervention in the surgical group was pulmonary regurgitation (64.9%) compared to the percutaneous group in which it was homograft/conduit stenosis or a mixed lesion (68.6%). RESULTS: There has been one (1.1%) early death reported in the surgical series and none in the percutaneous group. In the surgical group 9 (9.6%) experienced a procedural complication whilst 3 (8.5%) of those undergoing a percutaneous valve experienced a significant procedural event necessitating urgent surgery. Important early morbidity was 8 (8.5%) in the surgical group and 2 (5.7%) in the percutaneous group. Freedom from re-operation at 1 year was 100% in the surgical group and 86.1% in the percutaneous group due to late restenosis. Median hospital stay in the surgical group was 7 (4-114) days and 2 (2-22) days in the percutaneous group. CONCLUSIONS: Preliminary data suggests that percutaneous pulmonary valve implantation provides a promising additional and complementary approach to a successful surgical programme. Both approaches are safe with acceptable levels of morbidity and low mortality. With current technology the aneurysmal outflow tract remains a problem for the percutaneous approach. Follow-up remains too short, at present, to prove longevity of the percutaneous conduit. PMID- 15784348 TI - Intraoperative stenting of pulmonary arteries. AB - OBJECTIVE: The surgical treatment of pulmonary artery stenoses in small children with complex cardiac lesions can be technically difficult. A hybrid-approach combining corrective surgery and intraoperative stent placement may be complementary in these patients. METHODS: Descriptive study in 11 small children (age: one week to 12 years, median of 12 months, weight: 2.5-20 kg) after previous cardiac surgery. Intraoperative stenting of pulmonary arteries was performed involving paediatric cardiologist and cardiac surgeon. Stenting was combined with repair of pulmonary atresia (n=2), right ventricular outflow and pulmonary artery reconstruction (n=3), unifocalisation of pulmonary arteries (n=1), revision of distal anastomosis of RV-PA-conduit after truncus repair (n=1), revision of distal anastomosis of RV-PA-conduit after AVSD/Fallot repair (n=1), aortic arch patch reconstruction after anatomical correction for transposition of the great arteries (n=1), bidirectional cavopulmonary anastomosis after Norwood I operation for hypoplastic left heart syndrome, (n=1) retrieval of a dislodged stent from the left pulmonary artery (n=1). In seven patients stenting was planned electively while in four patients it took place on an emergency base. RESULTS: No complications occurred during stent implantation. One patient died three weeks postoperatively from diffuse bleeding due to a coagulation disorder. Ten patients left hospital after the surgical intervention with concomitant stent implantation. Stent re-dilatation was necessary in 4 patients 2-24 months postoperatively. After a median follow-up of 15 months ranging from 3 weeks to 7.5 years all stents were patent as diagnosed by angiography in 6 patients and by colour-Doppler in all other patients. One year after stent placement one stent was removed and another surgically opened during re-operation for conduit replacement in the smallest patient from this series. There was one late death during operative right ventricular outflow-tract reconstruction after initial stent placement. CONCLUSIONS: With intraoperative stent placement surgically difficult patch augmentation of small and fragile pulmonary vessels during repair of complex cardiac lesions can be avoided. Stents recruit pulmonary vessels and keep them open and amenable to future percutaneous or surgical interventions. PMID- 15784349 TI - Oversizing pulmonary homograft conduits does not significantly decrease allograft failure in children. AB - OBJECTIVE: Placement of oversized pulmonary ventricle-pulmonary artery conduits is routinely performed to decrease conduit failure in children. However, this practice has recently been challenged as somatic outgrowth may not be the main determinant of allograft failure in children. Our objective was to determine whether placement of oversized homografts for extracardiac pulmonary ventricle (PV) outflow tract reconstruction improves longevity in young children. METHODS: We reviewed 102 consecutive PV-PA conduits inserted in 70 patients less than 18 years between 1984 and 2003. Conduits placed in an anatomic position (n=23) as part of a Ross operation, were excluded. Conduits were initially stratified into two age groups: Group 1, those placed in patients 10 years. Normalization of conduit size to patient's body surface area at the time of insertion (z-value) was then performed to divide the conduits into oversized (O/S) and non-oversized (NO/S) groups. Determinants of conduit failure and allograft longevity were then compared between groups. RESULTS: Seventy-nine extracardiac conduits were placed, and 57 of these were in patients under 10 years of age. The majority had a diagnosis of tetralogy of Fallot (n=38), truncus arteriosus (n=19), pulmonary atresia with ventricular septal defect (n=12), or D-TGA with pulmonary stenosis and ventricular septal defect (n=7). Thirty-seven conduits were oversized (O/S) based on z-value, and 42 were non-oversized (NO/S), and the mean age at initial homograft placement was 7.0+/-7.5 years. Overall, oversizing conferred no significant advantage with respect to actuarial freedom from homograft replacement at 1, 5, or 10 years (96, 79, and 21%, O/S vs 93, 60, and 24%, NO/S), P=0.44. Oversizing was more frequent in Group 1 than Group 2 (53 vs 32%), and conduit failure was also more frequent with 49% requiring reoperation during the study period vs 38% in Group 2. In the subset of patients 0.2), while the diastolic chamber stiffness constant (k(ED)) significantly increased (0.014+/-0.005 to 0.040+/-0.007 ml-1, P=0.018) and relaxation time constant (tau) significantly decreased (61+/-3 to 49+/-2 ms, P=0.004). Before CPB, incremental atrial pacing had no significant effects on E(ES) and tau but significant negative effects on kED (0.014+/-0.005 to 0.045+/ 0.012 ml-1, P=0.013). After CPB, atrial pacing had significant positive effects on E(ES), tau and kED (E(ES): 1.12+/-0.28 to 2.60+/-1.54 mm Hg/ml, P=0.021; tau: 49+/-2 to 45+/-2 ms, P=0.009; kED: 0.040+/-0.007 to 0.026+/-0.005 mm Hg, P=0.010), indicating improved systolic and diastolic chronotropic responses. CONCLUSION: On-pump normothermic CABG with IAWBC preserved systolic function, increased diastolic stiffness, and improved systolic and diastolic chronotropic responses. Normalization of the chronotropic responses post-CPB is likely due to effects of successful revascularization and subsequent relief of ischemia. PMID- 15784358 TI - Is cardiopulmonary bypass a reason for aspirin resistance after coronary artery bypass grafting? AB - OBJECTIVE: 'Off-pump' coronary artery bypass grafting (OPCAB) is an alternative to conventional coronary artery bypass grafting (CABG) using cardiopulmonary bypass (CPB). While midterm results after OPCAB have become available, systematic studies of changes in platelet function after OPCAB are still missing. Since we have previously shown that oral aspirin treatment (100mg) does not achieve sufficient platelet inhibition in the majority of patients operated on with CPB, we hypothesized that bypass surgery without CPB (off-pump coronary artery bypass, OPCAB) causes less impairment of platelet inhibition by aspirin. The aim of this study was to investigate platelet function and the antiplatelet effect of aspirin after off-pump coronary artery bypass grafting in comparison with conventional on pump surgery. METHODS: We compared platelet function (in vitro aggregation and thromboxane formation) before and at days 1 and 5 after coronary artery bypass grafting, performed with (n=15) or without (n=14) CPB. Oral aspirin treatment (100mg/d) was started at day 1 after surgery. RESULTS: After a 5 day oral treatment with aspirin, platelet aggregation was inhibited significantly in OPCAB patients to 55.7+/-16.3% of control before surgery (P<0.05), whereas aggregation remained unchanged after CPB (105.8+/-26.9% of control before surgery; P>0.05). Since aspirin primarily inhibits platelet thromboxane formation, thromoboxane was determined after in vitro aggregation. According to platelet aggregation, thromboxane formation was only inhibited significantly after OPCAB (29.2+/-13.0% of control before surgery, P<0.05), but not after CPB (74.5+/-21.4% of control before surgery, P>0.05). This resistance to aspirin after CPB may be caused by an increased release of new platelets which are competent to form thromboxane, since the number of platelets decreased from 237+/-11x10(3)/microl before CPB to 174+/ 13x10(3)/microl at day 1 after surgery and increased significantly the following days reaching 303+/-17x10(3)/microl at day 5. Platelet counts of patients operated on without CPB showed no significant changes (236+/-16x10(3)/microl before OPCAB, 220+/-16x10(3)/microl at day 1 and 266+/-31x10(3)/microl at day 5 after surgery). CONCLUSIONS: The antiplatelet effect of aspirin is largely impaired after CPB, but not after CABG without CPB. Hence, increased platelet turnover after CPB seems to contribute to aspirin resistance, since an increased number of platelets might be competent to form thromboxane within the dosing intervals. PMID- 15784359 TI - Antioxidant defence during cardiopulmonary bypass surgery. AB - OBJECTIVE: Cardiac surgery may lead to severe oxidative stress due to formation of oxidation products generated during ischemia and reperfusion. We investigated to which extent oxidative stress influences a number of endogenous antioxidants and markers of cellular activation. METHODS: At six time points blood was withdrawn from patients undergoing coronary artery bypass grafting, using the on pump procedure. RESULTS: Both glutathione peroxidase and superoxide dismutase show a gradual and strong increase in activity during surgery (40 and 30%, respectively), returning to baseline values 24 h after surgery. The total antioxidant capacity has a maximum increase of 60%. Markers of cellular activation, such as eosinophil cationic protein and tryptase also increase during the procedure. CONCLUSION: Cardiac surgery results in systemic inflammation accompanied or caused by severe oxidative stress. The human body has a strong innate oxidative defence screen, which is probably not sufficient to fully compensate for the total amount of oxidative damage. PMID- 15784360 TI - The GP IIb/IIIa inhibitor abciximab (ReoPro) decreases activation and interaction of platelets and leukocytes during in vitro cardiopulmonary bypass simulation. AB - OBJECTIVE: Cardiopulmonary bypass (CPB) induces a systemic inflammatory response and increases expression of the platelet activation marker P-selectin which mediates binding of platelets to leukocytes. Inhibition of the platelet GP IIb/IIIa receptor during CPB has been shown to protect platelets without increasing bleeding complications and is assumed to reduce the inflammatory response. The aim of this study was to investigate the effect of the GP IIb/IIIa inhibitor abciximab (ReoPro) on the function and interaction of platelets and leukocytes during experimental CPB. METHODS: Heparinized (3 U/ml) fresh whole blood of healthy volunteers was treated before continuous in vitro circulation in a well established CPB model with 3.2 microg/ml abciximab (n=6) or left untreated as control (n=6). Measurements were made before (baseline) and after 30 and 60 min of circulation and comprised: percentage of platelets expressing P-selectin and percentage of platelet-bound leukocytes (flow cytometry), release of the leukocyte activation marker PMN-elastase (ELISA), and platelet and leukocyte counts. RESULTS: Abciximab almost completely prevented a CPB-induced increase of platelet P-selectin and platelet-leukocyte binding after 30 and 60 min of circulation, and significantly inhibited release of PMN-elastase after 30 min of circulation. Furthermore, abciximab significantly inhibited a CPB-induced decrease of platelet and leukocyte counts. CONCLUSIONS: Abciximab inhibits CPB induced activation, interaction and consumption of platelets and leukocytes in vitro. GP IIb/IIIa inhibition should be considered as a promising approach not only to conserve platelet function but also to inhibit pro-inflammatory events during CPB in vivo. PMID- 15784361 TI - Stroke in surgery of the arteriosclerotic descending thoracic aortic aneurysms: influence of cross-clamping technique of the aorta. AB - OBJECTIVE: The risk of stroke caused by dislodgment of loose atheromatous plaque or mural emboli is increased by cross-clamping of the aorta. Some patients undergo descending thoracic aortic aneurysm repair with proximal aortic cross clamping between the left common carotid artery and the left subclavian artery. The objective of this study was to determine the influence of proximal aortic cross-clamping in arteriosclerotic aneurysm or dissecting aneurysm repair. METHODS: Between May 1984 and May 2003, 81 patients underwent elective surgery for distal arch or descending aortic aneurysm repair with proximal aortic cross clamping between the left common carotid artery and the left subclavian artery. To evaluate the influence of the proximal aortic cross-clamping, patients were divided into two groups: patients who had undergone arteriosclerotic aneurysm repair (group I, n=25) and patients who had undergone dissecting aneurysm repair (group II, n=56). RESULTS: Eight (9.9%) of the 81 patients had a stroke. Six strokes occurred in operations for arteriosclerotic aneurysm repair group I and two strokes occurred in operations for dissecting aneurysm repair group II (24 vs 3.6%; p=0.009). In-hospital mortality rates were 12% in group I and 8.9% in group II (p=0.70). Major postoperative complications included renal failure requiring hemodialysis (in 4.2% of the patients in group I and in 8.3% of the patients in group II, p=0.99) and pulmonary complication (in 20% of the patients in group I and in 16% of the patients in group II, p=0.67). CONCLUSION: Cross-clamping between head vessels should be avoided if at all possible when operating on patients who have arteriosclerotic descending thoracic aneurysms. PMID- 15784362 TI - Composite aortic root replacement in acute type A dissection: time to rethink the indications? AB - OBJECTIVE: The indications for aortic root replacement in acute type A dissection are unclear. We reviewed the immediate and long-term outcome of consecutive patients in a series in which a low-threshold policy of composite aortic root replacement had evolved. METHODS: From a prospectively compiled aortic surgery database, we identified 162 patients who had either supracoronary interposition grafting, Group A (n=89), or composite root replacement, Group B (n=73) for acute type A dissection. Patients receiving total arch replacements were excluded. Operative and clinical details were analyzed and patient survival was compared to an age and gender matched census cohort. Need for reoperation on the proximal or distal aorta was also noted. Follow-up totaled 795.5 patient-years. RESULTS: Hospital mortality rates were identical in both groups (12.3%: 11 deaths in group A; 9 in group B). Chronic pulmonary disease, diabetes, malperfusion, hemodynamic compromise and aortic root dilatation were independent risk factors for hospital death. Actuarial survival estimates at 1, 5 and 10 years were 79% (71-88%), 64% (53-75%), and 55% (41-68%) for group A, and 79% (70-86%), 73% (62-83%), and 65% (52-78%) for group B (P=0.48). Age and operative patency of the ascending false lumen were independent risk factors for death after hospital discharge. Proximal aortic reoperation was required for four patients in group A and none in group B (P=0.085). CONCLUSION: A strategy of replacement rather than repair of the dissected aortic root for specific indications in type A dissection yielded high survival and low proximal reoperation rates. These results support an aggressive policy of composite root replacement in acute type A dissection. PMID- 15784364 TI - Technical problems and complications of axillary artery cannulation. AB - OBJECTIVE: Cardiopulmonary bypass via the axillary artery is frequently used especially in aortic dissections. With an increased use of this technique problems were recognized too. We describe the technical problems and complications associated with axillary artery cannulation. METHODS: Sixty-five patients underwent cannulation of the axillary artery. The indication for operation was acute aortic dissection type A in 57%, chronic aortic dissection in 8%, aortic aneurysm in 18%, pseudoaneurysm in 3%, and others in 14%. RESULTS: Technical problems and complications occurred in 14%, and in 11% the perfusion had to be switched to either femoral (n=5) or aortic cannulation (n=2). Arterial damage or dissection of the axillary artery or the aorta occurred in 0% of the sidegraft technique, whereas they were found in 9% with direct cannulation (P=n.s.). Cannulation problems or insufficient CPB flow due to a narrow vessel occurred in 0% of the sidegraft technique, whereas they were found in 4% with direct cannulation (P=n.s.). Malperfusion in aortic dissections occurred in 20% of the sidegraft technique, whereas they were found in 0% with direct cannulation (P=0.016). No postoperative complications related to axillary cannulation which were evaluated by clinical examination, such as brachial plexus injury, axillary artery thrombosis or local wound infection were observed. CONCLUSIONS: Although axillary artery cannulation is an attractive alternative to femoral cannulation there needs to be an alertness for technical problems. Different complications occur with either direct cannulation or the sidegraft technique and at present it remains the surgeons preference which technique for axillary artery cannulation is used. PMID- 15784365 TI - Arch repair with unilateral antegrade cerebral perfusion. AB - OBJECTIVE: Several antegrade cerebral perfusion techniques with differing neurological outcomes are employed for aortic arch repair. This study demonstrates the clinical results of aortic arch repair with unilateral cerebral perfusion via the right brachial artery. METHODS: Between January 1996 and March 2004, 181 patients underwent aortic arch repair via the right upper brachial artery with the use of low-flow (8-10 ml/kg per min) antegrade selective cerebral perfusion under moderate hypothermia (26 degrees C). Mean patient age was 58+/-12 years. Presenting pathologies were Stanford type A aortic dissection in 112, aneurysm of ascending and arch of aorta in 67, and isolated arch aneurysm in two patients. Ascending and/or partial arch replacement was performed in 90 patients and ascending and total arch replacement in 91 patients (including 27 with elephant trunk). In a subset of patients, renal and hepatic effects of ischemic insult were assessed. Free hemoglobin and lactate dehydrogenase levels were measured pre and postoperatively to identify hemolytic effects of brachial artery cannulation. RESULTS: Mean antegrade cerebral perfusion time was 36+/-27 min. Three patients with acute proximal dissection died due to cerebral complications. One patient had transient right hemiparesis. Total major neurological event rate was 2.2%. Brachial artery was able to carry full cardiopulmonary bypass flow with mild hemolysis. Renal and hepatic tests showed no deleterious effects of limited ischemia at moderate hypothermia. CONCLUSIONS: Arch repair with antegrade cerebral perfusion through right brachial artery has excellent neurological results, provides technical simplicity and optimal repair without time restraints, does not necessitate deep hypothermia and requires shorter CPB and operation times. PMID- 15784366 TI - Malposition of selective cerebral perfusion catheter is not a rare event. AB - OBJECTIVE: Although malposition of a catheter for selective cerebral perfusion can lead to postoperative neurologic complications, the clinical relevance or even an incidence of this event is not clear because there have been no measures to diagnose it. The purpose of this study is to report the results of intraoperative diagnosis of catheter malposition by means of near-infrared spectroscopy, orbital ultrasound, and transesophageal echocardiography. METHODS: The 35 consecutive patients of aortic arch aneurysm undergoing total arch replacement (13 patients) or transaortic stent graft implantation (22 patients) were examined. The regional oxygen saturation in the frontal lobe was continuously monitored with near-infrared spectroscopy. When cerebral malperfusion was suspected with saturation drop and reduced blood flow in orbital ultrasound, blood flow in the cervical branches and catheter position were examined with transesophageal echocardiography. RESULTS: Catheter malposition was detected in 4 of 35 cases (11.4%). The echo findings included: (1) reduced or absent flow and/or collapsed lumen in the common carotid artery despite an adequate perfusion rate; and (2) the balloon of catheter blocking the inflow to the common carotid artery. There was no unusual changes in parameters of other conventional monitors. After the catheter was withdrawn (three cases) or replaced (one case) based on the above diagnosis, cerebral perfusion was restored, confirmed by these three modalities. An accidental entry of catheter into the right common carotid artery was detected by transesophageal echocardiography in one case, in which there was no abnormal finding of oxygen saturation or orbital blood flow. CONCLUSIONS: Catheter malposition on the right side is not a rare event during selective cerebral perfusion. The catheter can migrate into the right subclavian artery or common carotid artery. Pressure monitoring cannot reliably detect an occurrence of catheter migration into the right subclavian artery. Combined use of near-infrared spectroscopy, orbital ultrasound, and transesophageal echocardiography can be useful for detecting this event and making an appropriate decision without delay to prevent irreversible brain damage. PMID- 15784367 TI - Stent grafting technique using Matsui-Kitamura (MK) stent for patients with aortic arch aneurysm. AB - OBJECTIVE: Although several methods of stent-grafting for patients with aortic arch aneurysm have been reported, these methods have been associated with several complications such as endoleak and migration. We developed a new method using Matsui-Kitamura (MK) stent-graft following extra-anatomic arch vessels bypass by selective cerebral perfusion (SCP) under left heart bypass (LHB). METHODS: Between December 2001 and December 2003, 12 patients with aortic arch aneurysm were treated according to this new method. All patients were male with an average age of 71.3+/-6.4 years. There were 5 patients with severe pulmonary dysfunction, 4 with renal dysfunction, one with severe cardiac dysfunction and 3 with preoperative cerebral infarction. Under SCP using LHB, the extra-anatomic arch vessel bypass was established. The MK stent-graft was delivered into the aortic arch. Coronary artery bypass grafting (CABG) was concomitantly performed in one patient. RESULTS: There were no cases of endoleak, migration or hospital death. One patient, who had a past history of cerebrovascular disease, suffered a minor stroke, and one patient, who was performed CABG to the mid-left anterior descending branch (LAD) using the left internal thoracic artery (LITA), presented paraparesis. Although two patients of chronic renal failure underwent scheduled CHDF on account of using the contrast medium during the procedure, all of them were weaned from hemodialysis. However, there were no other postoperative complications such as, respiratory failure or cardiac dysfunction. CONCLUSIONS: Endovascular stent grafting EVSG using the MK stent with extra-anatomic arch vessel bypass under SCP using LHB could be a useful and less invasive method for patients with aortic arch aneurysm who are at a high surgical risk. PMID- 15784368 TI - Risk factors for secondary dilatation of the aorta after acute type A aortic dissection. AB - OBJECTIVES: Prompt diagnosis of subsequent dilatation of the dissected aorta is crucial to reduce late mortality in these patients. This study focuses on risk factors for dilatation of the aorta after type A aortic dissection (AADA) affecting a normal-sized or slightly dilated aorta. METHODS: Overall 531 CT scans were analysed. Patients were included in the study if at least 3 CT scans were available after operative repair. 64 patients (59.8%) out of 107 patients full field the inclusion criteria. Volumetric analyses of the aorta were performed. Patients were divided in 3 groups: group A included 26 patients (40.6%) without progression of the aortic diameter, group 2, 27 patients (42.2%) with slight progression and group 3, 11 patients (17.2%) with important progression, requiring surgery in 9 patients (81.8%). Risk-factors for progression of the aortic size were analysed and compared between the groups. RESULTS: Patients from group 3 were younger 57.7+/-13.4 vs. 61.9+/-11.6 in group 1 (P<0.05) and were more frequent female (45.4 vs. 23.1%; P<0.05). Dissection of the supraaortic branches (100 vs. 80.8%; P<0.05), the presence of preoperative cerebral, visceral or peripheral malperfusion (54.6 vs. 26.9%; P<0.05) and contrast enhancement in the false lumen during the follow-up (72.7 vs. 57.7%; P=0.07) were additional risk factors for late aortic dilatation in these patients. CONCLUSIONS: Acute type A aortic dissection in younger patients, involving the supraaortic branches and/or combined with malperfusion syndrome favour secondary dilatation. A close follow-up is mandatory to prevent acute complications of the diseased downstream aorta following repair of a AADA. PMID- 15784369 TI - Treatment of congenital cystic adenomatoid malformation-does resection in the early postnatal period increase surgical risk? AB - OBJECTIVE: The recent development of fetal ultrasonography has allowed for an increasing number of prenatal diagnoses for congenital cystic adenomatoid malformation (CCAM). However, the appropriate surgical timing of these patients has not been studied as of yet. The aim of this study is to suggest a safe strategy for the treatment of CCAM by identifying the relationship between the timing of surgery and postoperative outcome. METHODS: Between 1987 and 2003, 40 patients (28 males, 12 females) underwent surgical resection for CCAM. The mean age was 38.6+/-9.1 (2 days-13 years) months. CCAM was diagnosed by prenatal ultrasonography in eight patients. Early operations were performed in four out of the eight. Operation was deferred until 2-12 months of age for the remaining four patients. RESULTS: Type I CCAM was found in 20 patients, type II in 20 and no patient exhibited type III. Five patients had associated pectus excavatum anomaly. There were no cases of operative mortality. Seventeen minor postoperative complications developed in 16 patients (40.0%): prolonged chest tube drain in 10, wound infection in 4, and 1 case of pneumonia, empyema and pleural space, respectively. The average hospital stay was 11.8 (6-29) days. During the mean follow-up period of 67.5 months, one patient died of accidental aspiration 7 months after operation during the postoperative recovery course of Ravich operation for pectus excavatum. The remaining patients reported doing well with normal physical activity. All five patients who underwent surgery at the age of under 1 month did not exhibit increased postoperative morbidity. CONCLUSIONS: We concluded that surgery for CCAM could be safely performed in all age groups with satisfactory long-term outcomes. It is suggested that early elective surgical correction can be recommended for a patient whose diagnosis was made in utero. PMID- 15784370 TI - Catamenial pneumothorax: optimal hormonal and surgical management. AB - OBJECTIVE: To provide further information addressing the etiology, optimal hormonal management and surgical management in catamenial pneumothorax (CP). METHODS: We retrospectively analyzed records of all female patients operated on for spontaneous pneumothorax at a university hospital between January 1993 and March 2002. RESULTS: In eight of 24 patients, pneumothoraces were timed with menses. In all, the right side was involved. Seven patients were on hormonal medications pre-operatively and six post-operatively. All six patients taking estrogen/progesterone replacement had recurrences pre-operatively and two of three had recurrences post-operatively while on these medications. No patient suffered a pneumothorax either pre- or post-operatively while taking a gonadotropin releasing hormone agonist (two and three patients, respectively). Intraoperative findings included diaphragmatic implants [5] diaphragmatic fenestrations [4], apical blebs [2] and visceral pleural implants [2]. All pathology was specifically addressed at the time of surgery. Pleural space management included mechanical pleurodesis in seven and pleurectomy with talc insufflation in 1. Follow-up ranged from 27 to 63 months with a mean of 48 months. Three patients developed post-operative recurrences. One was managed without intervention and two required additional procedures. CONCLUSION: Catamenial pneumothorax is under appreciated, representing up to one-third of women with spontaneous pneumothorax. Hormonal agents that allow for menses are ineffective. Gonadotropin releasing hormone agonists should be considered as part of the pre-operative or post-operative management in high risk patients. Our findings suggest that an additional intervention to augment pleural symphysis at the level of the diaphragm should be performed. PMID- 15784371 TI - Use of intrapleural streptokinase in experimental minimal clotted hemothorax. AB - OBJECTIVE: In clotted hemothorax, both thoracocentesis and closed tube thoracostomy will not be able to evacuate the pleural cavity especially if it is minimal. The aim of this study was to assess the effectiveness of intrapleural administered streptokinase on minimal clotted hemothorax without drainage, in order to accelerate the spontaneous resolution and absorption in blunt thoracic trauma. METHODS: Thirteen adult ewes were used for this experiment. The animals were divided into two groups. First group served as the control group (Group C) (n=5) and did not receive any intrapleural fibrinolytic treatment. In both groups, 200 ml of blood was taken from the left jugular vein and injected into the pleural cavity with a serum line through the scope after pleural abrasion. Streptokinase (150.000 U) was diluted in 100 ml of saline and applied to the second group (Group S) (n=5) in second postoperative day. One ewe in each group was sacrificed with a lethal dose of sodium thiopental in postoperative 2nd, 4th, 6th, 8th, and 10th weeks, respectively. When a left posterolateral thoracotomy was performed, pleural thickening and adhesion were evaluated. The lung and pleural tissue samples were taken for histopathologic examination. The slides were examined in a blinded manner. RESULTS: Thoracentesis was performed in all ewes in the second postoperative day and no fluid was detected. There was no allergic reaction in group S after the injection of streptokinase into the pleural cavity. During postmortem macroscopic evaluation, we observed clot in one of the ewes in group C in second postoperative week. A statistically significant difference was found between Group C and S regarding pleural thickening and adhesion (P=0.05). The ewes of Group S had less pleural thickening and adhesion compared to those of Group C. These results were confirmed with histopathological examination. CONCLUSION: We conclude that intrapleural streptokinase increases resolution of clot in the pleural space and decreases pleural thickening and adhesion in experimental minimal clotted hemothorax in ewes. This study has also demonstrated that intrapleural streptokinase can be used without drainage. Use of intrapleural streptokinase without drainage can be a novel therapeutic option for trauma patients with minimal clotted hemothorax after haemorrhage of other organs was excluded. PMID- 15784372 TI - Does a thoracic epidural confer any additional benefit following video-assisted thoracoscopic pleurectomy for primary spontaneous pneumothorax? AB - OBJECTIVE: Video-assisted thoracoscopic (VATS) bullectomy and apical pleurectomy has become the preferred procedure for recurrent or complicated primary spontaneous pneumothorax (SPN). Although thoracic epidural analgesia is the gold standard after open thoracic surgical procedures, its use in the management of minimally invasive procedures in this young population has not been extensively studied. METHODS: From 1997 to 2003, a single surgeon performed 118 consecutive VATS pleurectomies for primary SPN. The perioperative course, analgesic requirements, hospital stay and long-term complications were compared for 22 (18%) patients in whom a patient-controlled thoracic epidural was used for analgesia and 96 (82%) patients who did not receive an epidural (parenteral opioids). A four-point verbal pain score (0-3) was recorded hourly in every patient at rest and on coughing following surgery. RESULTS: One patient required additional surgery for evacuation of haemothorax. There were no mortalities or other major complications in the series. Overall median hospital stay was 3 (range 1-10) days, the incidence of long-term pain at 3 months was 6%, and the long-term recurrence rate was 3%. Despite parenteral opioids being discontinued significantly earlier than epidurals, pain scores were similar in both groups. There were no significant differences in the duration of air-leaks, length of drainage, hospital stay, long-term pain and long-term paraesthesias between the two groups. CONCLUSIONS: Thoracic epidural analgesia does not contribute significantly to minimize neither perioperative nor long-term pain after VATS pleurectomy for primary SPN. The additional resource requirement in these patients is not justified. PMID- 15784373 TI - A case-matched study of anatomical segmentectomy versus lobectomy for stage I lung cancer in high-risk patients. AB - OBJECTIVE: Sublobar resections may offer a method of increasing resection rates in patients with lung cancer and poor lung function, but are thought to increase recurrence and therefore compromise survival for stage I non-small cell lung cancer (NSCLC). To test this hypothesis we have compared the long-term outcome from lobectomy and anatomical segmentectomy in high-risk cases as defined by predicted postoperative FEV1 (ppoFEV1) less than 40%. METHODS: Over a 7-year period 55 patients (27% of all resections for stage I NSCLC) with ppoFEV1<40% underwent resection of stage I NSCLC. The 17 patients who underwent anatomical segmentectomy were individually matched to 17 patients operated by lobectomy on the bases of gender, age, use of VATS, tumour location and respiratory function. We compared their perioperative course, tumour recurrence and survival. RESULTS: There were no significant differences in hospital mortality (one case in each group), complications or hospital stay. Overall 5-year survival was 69%. There were no differences in recurrence rates (18% in both groups) or survival (64% after lobectomy and 70% after segmentectomy). There was preservation of pulmonary function after segmentectomy (median gain of 12%) compared to lobectomy (median loss of 12%) (P=0.02). CONCLUSIONS: Anatomical segmentectomy allowed for surgical resection in patients with stage I NSCLC and impaired respiratory reserve without compromising oncological results but with preservation in respiratory function. PMID- 15784374 TI - Does the extent of lymph node dissection influence outcome in patients with stage I non-small-cell lung cancer? AB - OBJECTIVE: To assess the therapeutic effect of the extent of lymph node dissection performed in patients with a stage pI non-small-cell lung cancer (NSCLC). METHODS: We analysed data on 465 patients with stage I NSCLC who were treated with surgical resection and some form of lymph node sampling. The median number of lymph node sampled was 10 and the median number of ipsilateral mediastinal lymph node stations sampled was two. We chose to define a procedure that harvested 10 or more lymph nodes and sampled two or more ipsilateral mediastinal stations as a lymphadenectomy, by contrast with sampling when one or both criteria were not satisfied. The effect of the surgical techniques: lymph node sampling (LS; n=207) vs. lymphadenectomy (LA; n=258) on 30-day mortality and overall survival were investigated. RESULTS: A total of 6244 lymph nodes was examined, including 4306 mediastinal lymph nodes. The mean (+/-SD) numbers of removed lymph nodes were 7+/-6.1 per patient following LS vs.18.6+/-9.3 following LA (P=0.001). An average mean of 1+/-0.90 mediastinal lymph node station per patient was sampled following LS vs. 2.7+/-0.8 following LA (P<10(-6)). Overall 30-day mortality rates were 2.4 and 3.1%, respectively. LA was disclosed as a favourable prognosticator at multivariate analysis (Hazard Risk: 1.43; 95% Confidence Interval: 1.00-2.04; P=0.048), together with younger patient age, absence of blood vessels invasion, and smaller tumour size. CONCLUSIONS: Importance of lymph node dissection affects patients outcome, while it does not enhance the operative mortality. A minimum of 10 lymph nodes assessed, and two mediastinal stations sampled are suggested as possible pragmatic markers of the quality of lymphadenectomy. PMID- 15784375 TI - Prognosis and survival after radical resection of primary adenosquamous lung carcinoma. AB - OBJECTIVE: In order to evaluate the follow-up study of surgical treatment for primary adenosquamous lung carcinoma (ASC) we specified prognostic criteria, also in comparison with primary adenocarcinoma (AC). METHODS: The study group consisted of 96 patients discharged between 1990 and 1999 after radical surgical treatment for ASC-80 (83%) men and 16 (17%) women aged 34-73, mean 56 years. Consequently, we evaluated 252 patients operated during the same time period for primary AC. RESULTS: Apart from grading, we did not find any significant differences between both ASC and AC groups of patients. Among the 96 patients operated radically for ASC median overall survival (OS) was 20 months. The cumulative postoperative survival rates at 5 and 10 years were 25.4 and 19.2%, respectively. By comparison, median OS for 252 patients with AC, discharged after surgical treatment in the same period, was 28.5 months and the cumulative postoperative survival rates at 5 and 10 years were 42.5 and 39.1%, respectively (P=0.006). At pathologic stages IA, the cumulative survival rate at 5 years was 63.3% for patients with ASC as compared with 72.1% for patients with AC (P=0.330). However, out of IB stage patients treated surgically for ASC 31.8% survived 5 years in comparison with 56.3% operated for AC (P=0.017). Study of survival rates did not differ significantly between ASC and AC patients at stage IIA (P=0.824) and stage IIB (P=0.217), respectively. Univariate analysis revealed that six factors of tumor size, T status, N status, as well visceral pleura involvement, tumor localization (central vs. peripheral) and tumor structure were significantly associated with the survival rate according to these variables. Multivariate analysis using Cox's proportional hazards model indicated that T factor, nodal involvement and one of the tumor components predominating were significant factors associated with the postoperative survival of patients with ASC. CONCLUSIONS: Our findings indicate that in patients after radical operation for ASC, predominance for one of the histopathological components (adenous or squamous) within primary tumor is attended by worst prognosis. Our study confirmed also that the prognosis of ASC of the lung was poorer than that of primary AC. Lack of generally accepted diagnostic criteria and unclear prognosis, even in the pathologic stage I suggest that there is a need for prospective studies in this respect. PMID- 15784376 TI - Resistance to pressure of the stump after mechanical stapling or manual suture. An experimental study on sheep main bronchus. AB - OBJECTIVE: In a previous experimental study on 60 freshly slaughtered pig trachea, a statistically significant better resistance to pressure was found after mechanical stapling compared to hand suture. The objective of this study was to determine the resistance to pressure of a bronchial stump depending upon the closure technique (manual vs. mechanical) used in sheep 14 days after pneumonectomy. METHODS: Pneumonectomy was performed on 30 sheep, which were alternatively closed either by a double-layer running suture at 90 degrees to the cartilaginous rings or with an automatic stapling device. Exactly 14 days after pneumonectomy, the animals were sacrificed and the trachea with the bronchial stump was retrieved. Sutures were placed under pressure until air leakage was observed. The air-leakage pressure was recorded digitally. RESULTS: In both groups, there was no evidence of a bronchopleural fistula. As in the previous experimental study, mean values of air-leakage pressure revealed a large standard deviation in both groups (min. 0.16-max. 1.15 bar). Unlike the results in the first experiment there was no statistically significant difference between the two groups. CONCLUSIONS: After 14 days, when a bronchial stump is considered to be healed, the resistance to pressure of a mechanical suture is equal to that of the manual suture. PMID- 15784377 TI - Transthoracic esophagectomy for adenocarcinoma of the oesophagus: standard versus extended two-field mediastinal lymphadenectomy? AB - OBJECTIVE: Controversy continues over the optimal extent of lymphadenectomy for the surgical treatment of Adenocarcinoma of the oesophagus. METHODS: From 1996 to 2003, 102 transthoracic en-bloc esophagectomy were performed for adenocarcinoma. Based on the 1994 consensus conference of the International Society of Disease of Esophagus, 35 patients underwent standard lymphadenectomy whereas 67 underwent extended lymphadenectomy. Mortality, morbidity and long-term survival were reviewed in each group. RESULTS: Extended lymphadenectomy increased the number of resected lymph nodes and improved the healthy/invaded lymph node ratio. It allowed to detect skip nodal metastasis in 36.4% of the N+ patients. Morbidity was higher following extended lymphadenectomy, with respect to pulmonary complications, and blood transfusions requirement (P=0.04). However, operative mortality was similar in both groups (9 vs. 11%). Overall disease-free survival was 28% at 5 years. Median of survival was higher in N0 than in N+ patients (55 months vs. 20 months; P=0.02). Extended lymphadenectomy was associated with an improving of disease-free survival when compared to standard lymphadenectomy (41 vs. 10% at 5 years; P<0.05), especially in the subgroup of patients with a N0 disease (median of survival 44 months vs. 17 months; P=0.001). Based on multivariable analyses, predictive factors of recurrence affecting disease free survival were the pT status (P=0.02), standard lymphadenectomy (P=0.05) and extracapsular lymph node involvement (0.04). CONCLUSIONS: These results indicate that extended 2-field lymphadenectomy is an important component of the surgical treatment of patients with adenocarcinoma of the oesophagus. It increases the likelihood of proper staging and affects patient outcome, while it does not enhance the operative mortality. However, extended lymphadenectomy increases non fatal morbidity, especially the incidence of pulmonary complications and the need for blood transfusion. PMID- 15784378 TI - Fixed and dynamic airway obstruction in polychondritis. PMID- 15784379 TI - Tracheal compression by esophageal mucocele after surgical exclusion of the esophagus. PMID- 15784380 TI - Pitfall in the computed-tomography-diagnosis of postcardiotomy infection: iodine accumulation after irrigation mimicking retrosternal abscess. PMID- 15784382 TI - Plug of aortic vegetation into left main coronary artery. PMID- 15784383 TI - Giant, free flotating tendinous cord in the left ventricle. PMID- 15784384 TI - Ruptured mycotic aortic pseudoaneurysm with concomitant aorto-tracheal and aorto oesophageal fistulas. PMID- 15784385 TI - Protruding coronary stent. PMID- 15784386 TI - A left ventricular pseudoaneurysm due to mitral valve replacement. PMID- 15784387 TI - Closure of left ventricle perforation with the use of muscular VSD occluder. AB - Growing experience in interventional cardiology leads to the use of large diameter of vascular equipment. In some instances, the so-called hybrid procedures are performed. After performing the interventional procedure, the opening in ventricular wall is closed surgically. Our intention was to check if the MVSDO can be used to close the perforation in the heart after the interventional cardiology procedure performed through the left ventricular (LV) free wall. In three pigs under general anesthesia, the heart was exposed through a small substernal incision. The LV was punctured and an 18F sheath was introduced into the LV. A 14 mm MVSDO was inserted through the 10F Delivery System. Using both the echocardiographic and angiographic guidance, the MVSDO was placed on the LV wall to close the opening in the LV. Time and volume of bleeding was recorded. In all cases the occluder was successfully placed closing the opening, bleeding observed after deployment of occluder lasted for approximately 2 min. We think MVSD occluder can be used to close the LV free wall perforation after hybrid interventional cardiac procedure. Early bleeding through MVSDO might be resolved by the manufacturing of new occluder with better sealing properties. PMID- 15784388 TI - Aortic root substitution after aortic valve replacement: a prosthesis-sparing operation. AB - Patients who underwent isolated aortic valve replacement could come to attention for new onset aortic disease or progression of borderline alterations not corrected at the first operation, especially in the subset of bicuspid valve disease. We describe our technique in redo operations for aortic root disease, using only a vascular graft and sparing the previously implanted valve prosthesis. In case of normally functioning mechanical prosthesis, we always left the valve in situ and substituted the aortic root with a Dacron conduit, extending the replacement if necessary to the other diseased portions of the thoracic aorta. PMID- 15784389 TI - Pericardial sewing-cuff: an alternative surgical technique for full-root implantation of aortic allograft and pulmonary autograft. AB - The following is a description of a modified technique for the reinforcement of proximal anastomosis regarding the process of aortic allograft or pulmonary autograft full root implantation. A double width pericardial strip is used to create a pericardial sewing-cuff to be used as a neo-ring. This technique is said to be best suited for decreasing the operative bleeding, reducing anastomotic pitfalls and concomitant repair of the aortic annulus. PMID- 15784390 TI - Near fatal infection of a patient with a left ventricular assist device due to unrecognized fetal death. AB - We report on an unusual case of a young female patient who received an implantable LVAD after unsuccessful emergency coronary bypass surgery following acute myocardial infarction. After LVAD placement, it became evident that the patient had been pregnant. She had to undergo gynaecological surgery during mechanical support to remove the deceased fetus. PMID- 15784391 TI - Eight years survival after partial left ventriculectomy. AB - Remodelling the left ventricle by partial left ventriculectomy (PLV) has been proposed as an alternative surgical treatment for end-stage heart failure. We present the case of a patient with dilated idiopathic cardiomyopathy who underwent PLV and remains in NYHA functional Class I eight years after the surgery. PMID- 15784393 TI - Internal thoracic artery response to papaverine: takes time. PMID- 15784394 TI - Lower dose of heparin for cardiopulmonary bypass is not necessarily associated with lower drainage loss. PMID- 15784395 TI - Is congenital tricuspid insufficiency (CTI) as rare as it seems to be? PMID- 15784397 TI - The effect of single low-dose methylprednisolone on the cardiopulmonary bypass induced inflammatory response. PMID- 15784400 TI - Chest wall resection in patients with non-small cell lung cancer. PMID- 15784401 TI - Is lactate dehydrogenase level really a reliable predictor of pulmonary morbidity than other co-morbid parameters following lung resection for NSCLC? PMID- 15784404 TI - A combined small cell carcinoma of the lung containing three components: small cell, spindle cell and squamous cell carcinoma, revisited. PMID- 15784406 TI - Variability of serum levels of tumor necrosis factor-alpha, interleukin 6, and soluble interleukin 6 receptor over 2 years in young women. AB - Serum levels of tumor necrosis factor-alpha (TNF-alpha), interleukin 6 (IL-6), and soluble interleukin 6 receptor (sIL-6R) have been studied as risk factors of cardiovascular disease in longitudinal studies. However, it is unknown about their long-term intra-individual variations and whether single measurements of these cytokines and receptor are reliable biomarkers in epidemiological studies. In this study, serum levels of TNF-alpha, IL-6, and sIL-6R were assayed by ELISAs in 36 young, healthy women from whom three blood samples were collected at 12 month intervals over 2 years, and the intraclass correlation coefficients (ICC) were estimated. The ICC of 0.73 (95% CI=0.49-0.79) for TNF-alpha was comparable to those of other commonly used biomarkers, justifying its use in epidemiological studies. The ICC of 0.48 (95% CI=0.25-0.58) for IL-6 was not optimal. However, IL 6 has been demonstrated as a consistent risk factor for cardiovascular disease, suggesting it could still be a useful biomarker if its disease association is substantial. The ICC of 0.36 (95% CI=0.10-0.47) for sIL-6R was relatively low, and multiple samples would need to be collected in prospective studies for this receptor. PMID- 15784407 TI - Roles of reactive nitrogen intermediates and transforming growth factor-beta produced by immunosuppressive macrophages in the expression of suppressor activity against T cell proliferation induced by TCR stimulation. AB - The suppressor activity of splenic macrophages induced by Mycobacterium intracellulare infection (MI-M phi s) against T cell concanavalin A (Con A) mitogenesis is mediated by MI-M phi's mediators, such as reactive nitrogen intermediates (RNIs), phosphatidylserine, free fatty acids, prostaglandin E(2) and to a minor extent TGF-beta. Here, we have compared the roles of RNIs and TGF beta in the expression of MI-M phi's suppressor activity against Con A mitogenesis and anti-CD3 monoclonal antibody (mAb)- and anti-CD28 mAb-induced mitogenesis (TCR signal-induced mitogenesis) of the target T cells, and have found the following. First, N(G)-monomethyl-L-arginine (NMMA) inhibited MI-M phi's suppressor activity against TCR signal-induced mitogenesis as well as Con A mitogenesis. Second, anti-TGF-beta mAb weakly restored the MI-M phi-mediated suppression only in the case of Con A mitogenesis, under limited conditions, such as very low cell densities of MI-M phi s. Third, the blocking effects of NMMA plus anti-TGF-beta mAb were somewhat more prominent in the case of Con A mitogenesis than in the case of TCR signal-induced mitogenesis. Fourth, Con A- or TCR signal-stimulated MI-M phi s secreted significant amounts of the latent TGF beta but not the active one. These findings indicate that RNIs, but not TGF-beta, play important roles in the MI-M phi-mediated suppression of TCR signal-induced mitogenesis, as well as Con A mitogenesis, of the target T cells. PMID- 15784409 TI - Comparison of biological activity of recombinant woodchuck interferon gamma and tumor necrosis factor alpha produced in baculovirus and Escherichia coli expression systems. AB - The full-length cDNAs of recombinant woodchuck interferon gamma (rwIFN gamma) and woodchuck tumor necrosis factor alpha (rwTNF alpha) were cloned into baculovirus transfer vectors and expressed in insect Sf9 cells. The recombinant proteins secreted by the insect cells, bac-rwIFN gamma and bac-rwTNF alpha, were found to be functionally competent. Their biological activities were compared to those of rwIFN gamma and rwTNF alpha produced in the Escherichia coli (E. coli) expression system. The bac-rwIFN gamma demonstrated a 4.5-fold greater protective activity against encephalomyocarditis virus-induced cytolysis of woodchuck hepatocytes and that of class I MHC antigen presentation on the hepatocytes than rwIFN gamma derived from E. coli. The bac-rwTNF alpha was cytotoxic towards murine fibroblasts and able to upregulate class I MHC antigen display and these effects were about 18-fold greater than those triggered by rwTNF alpha from E. coli at a comparable protein level. In addition, the antiviral activity of bac-rwIFN gamma was inhibited by anti-wIFN gamma antibodies and the cytotoxicity of bac-rwTNF alpha neutralized by cross-reactive antibodies to murine TNF alpha. The study showed that the expression of rwIFN gamma and rwTNF alpha in the baculovirus system generated biologically active cytokines whose potency was considerably greater than those produced in E. coli. PMID- 15784408 TI - Circulating tumor necrosis factor alpha is higher in non-obese, non-diabetic Mexican Americans compared to non-Hispanic white adults. AB - Mexican Americans (MA) exhibit high risk for the insulin resistance syndrome characterized by subclinical inflammation and greater risk for type 2 diabetes compared with non-Hispanic white (NHW) adults. The reasons for this phenomenon remain obscure. Because the inflammatory cytokine, tumor necrosis factor-alpha (TNF alpha), is associated with insulin resistance in various models of obesity and diabetes, we sought to determine whether circulating concentrations of this cytokine and its soluble receptors are higher in MA than NHW, and also to determine if the TNF alpha system is related to the lower insulin sensitivity in MA. Fasting blood samples were used to determine concentrations of TNF alpha, soluble TNF receptors 1 (sTNFR1) and 2 (sTNFR2) in the same 13 MA (7 women, 6 men, age=27.0+/-2.0 years, BMI=23.0+/-0.7) and 13 NHW (7 women, 6 men, age=24.8+/ 1.5 years, BMI=22.8+/-0.6) previously shown to exhibit differences in insulin sensitivity. Circulating TNF alpha was significantly higher (3.11+/-0.38 vs. 2.10+/-0.24 pg/ml, p<0.05) and sTNFR2 was significantly lower (1324+/-85 vs. 1925+/-127 pg/ml, p<0.05) among MA compared with NHW subjects. Soluble TNFR1 was not different between groups (MA: 970+/-111 pg/ml vs. NHW: 1218+/-73 pg/ml, p=0.07). TNF alpha, sTNFR1 and sTNFR2 were not correlated with HOMA-IR when the two groups were analyzed in aggregate. This study documents higher circulating TNF alpha concentrations in non-obese, non-diabetic MA, a population group at increased risk for the metabolic syndrome and the untoward effects of sub clinical inflammation. The clinical implications of this difference, if any, are not yet known. PMID- 15784410 TI - Inter-individual and intra-individual variability in TNF-alpha production by human peripheral blood cells in vitro. AB - BACKGROUND: Two assays--isolated peripheral blood mononuclear cells (PBMC) and a whole blood assay (WBA)--are commonly used to study TNF-alpha production by an individual in order to distinguish between high and low cytokine producers. We assessed the reliability and reproducibility of these assays. METHODS: The PBMC assays (n=5) were performed weekly over a period of 6 weeks and the WBAs (n=4) weekly over a 4-week period. Polymethylmethacrylate particles (approx. 6 x 10(2) particles/cell) and optimal concentrations of endotoxin (6.25 and 12.5 ng/ml) were used as the stimulatory agents in PBMC and WBAs, respectively. TNF-alpha production was measured by ELISA. RESULTS: There was a high degree of both intra- and inter-individual variability of TNF-alpha secretion, with unpredictable changes in the amount of the cytokine produced by cells from the same donor. This variability could not be eliminated by correcting for cell numbers. CONCLUSION: The PBMC and WBA models of TNF-alpha production by human peripheral blood cells cannot be used for the evaluation of inter-individual variability in cytokine secretion due to the high intra-individual variability observed. In the case of PBMC this is partly due to differences in the confluency of the cells between individuals. PMID- 15784411 TI - Cytokine gene polymorphisms as potential risk and protective factors in renal cell carcinoma. AB - The major aim of this study was to investigate the association of the cytokine gene polymorphisms with the development of renal cell carcinoma (RCC). The study included 29 patients with RCC and 50 healthy controls. All genotyping (TNF-alpha, TGF-beta, IL-10, IL-6, IFN-gamma) experiments were performed using sequence specific primers PCR (PCR-SSP). It was found that TNF-alpha -308 G/G and TGF-beta codon 10-25 T/T-G/C genotypes were significantly higher in frequency in the patients with RCC group compared with the healthy control group. Additionally, the frequency of TNF-alpha -308 G allele was significantly higher in the patients when compared to controls. On the other hand, the frequencies of TNF-alpha -308 G/A, IL-6 C/C and TGF-beta1 codon 10-25 C/C-G/G genotypes were significantly lower in the cancer group compared with the healthy control group. However, after correction for multiple comparisons (Bonferroni), these results did not remain significant. Nevertheless, these findings suggest that the TNF-alpha -308 G/G and TGF-beta codon 10-25 T/T-G/C genotypes may be potential risk factors for RCC, whereas TNF-alpha -308 G/A, IL-6 C/C and TGF-beta1 codon 10-25 C/C-G/G genotypes may be possible protective factors. The number of the cases has to be increased to investigate the independency of these polymorphisms involved in the oncogenesis of RCC. PMID- 15784413 TI - Assessment of EEG synchronization based on state-space analysis. AB - Cortical computation involves the formation of cooperative neuronal assemblies characterized by synchronous oscillatory activity. A traditional method for the identification of synchronous neuronal assemblies has been the coherence analysis of EEG signals. Here, we suggest a new method called S estimator, whereby cortical synchrony is defined from the embedding dimension in a state-space. We first validated the method on clusters of chaotic coupled oscillators and compared its performance to that of other methods for assessing synchronization. Then nine adult subjects were studied with high-density EEG recordings, while they viewed in the two hemifields (hence with separate hemispheres) identical sinusoidal gratings either arranged collinearly and moving together, or orthogonally oriented and moving at 90 degrees . The estimated synchronization increased with the collinear gratings over a cluster of occipital electrodes spanning both hemispheres, whereas over temporo-parietal regions of both hemispheres, it decreased with the same stimulus and it increased with the orthogonal gratings. Separate calculations for different EEG frequencies showed that the occipital clusters involved synchronization in the beta band and the temporal clusters in the alpha band. The gamma band appeared to be insensitive to stimulus diversity. Different stimulus configurations, therefore, appear to cause a complex rearrangement of synchronous neuronal assemblies distributed over the cortex, in particular over the visual cortex. PMID- 15784412 TI - Processing of audiovisual speech in Broca's area. AB - We investigated cerebral processing of audiovisual speech stimuli in humans using functional magnetic resonance imaging (fMRI). Ten healthy volunteers were scanned with a 'clustered volume acquisition' paradigm at 3 T during observation of phonetically matching (e.g., visual and acoustic /y/) and conflicting (e.g., visual /a/ and acoustic /y/) audiovisual vowels. Both stimuli activated the sensory-specific auditory and visual cortices, along with the superior temporal, inferior frontal (Broca's area), premotor, and visual-parietal regions bilaterally. Phonetically conflicting vowels, contrasted with matching ones, specifically increased activity in Broca's area. Activity during phonetically matching stimuli, contrasted with conflicting ones, was not enhanced in any brain region. We suggest that the increased activity in Broca's area reflects processing of conflicting visual and acoustic phonetic inputs in partly disparate neuron populations. On the other hand, matching acoustic and visual inputs would converge on the same neurons. PMID- 15784414 TI - Investigations of dipole localization accuracy in MEG using the bootstrap. AB - We describe the use of the nonparametric bootstrap to investigate the accuracy of current dipole localization from magnetoencephalography (MEG) studies of event related neural activity. The bootstrap is well suited to the analysis of event related MEG data since the experiments are repeated tens or even hundreds of times and averaged to achieve acceptable signal-to-noise ratios (SNRs). The set of repetitions or epochs can be viewed as a set of independent realizations of the brain's response to the experiment. Bootstrap resamples can be generated by sampling with replacement from these epochs and averaging. In this study, we applied the bootstrap resampling technique to MEG data from somatotopic experimental and simulated data. Four fingers of the right and left hand of a healthy subject were electrically stimulated, and about 400 trials per stimulation were recorded and averaged in order to measure the somatotopic mapping of the fingers in the S1 area of the brain. Based on single-trial recordings for each finger we performed 5000 bootstrap resamples. We reconstructed dipoles from these resampled averages using the Recursively Applied and Projected (RAP)-MUSIC source localization algorithm. We also performed a simulation for two dipolar sources with overlapping time courses embedded in realistic background brain activity generated using the prestimulus segments of the somatotopic data. To find correspondences between multiple sources in each bootstrap, sample dipoles with similar time series and forward fields were assumed to represent the same source. These dipoles were then clustered by a Gaussian Mixture Model (GMM) clustering algorithm using their combined normalized time series and topographies as feature vectors. The mean and standard deviation of the dipole position and the dipole time series in each cluster were computed to provide estimates of the accuracy of the reconstructed source locations and time series. PMID- 15784415 TI - Evaluation of different cortical source localization methods using simulated and experimental EEG data. AB - Different cortical source localization methods have been developed to directly link the scalp potentials with the cortical activities. Up to now, these methods are the only possible solution to noninvasively investigate cortical activities with both high spatial and time resolutions. However, the application of these methods is hindered by the fact that they have not been rigorously evaluated nor compared. In this paper, the performances of several source localization methods (moving dipoles, minimum Lp norm, and low resolution tomography (LRT) with Lp norm, p equal to 1, 1.5, and 2) were evaluated by using simulated scalp EEG data, scalp somatosensory evoked potentials (SEPs), and upper limb motor-related potentials (MRPs) obtained on human subjects (all with 163 scalp electrodes). By using simulated EEG data, we first evaluated the source localization ability of the above methods quantitatively. Subsequently, the performance of the various methods was evaluated qualitatively by using experimental SEPs and MRPs. Our results show that the overall LRT Lp norm method with p equal to 1 has a better source localization ability than any of the other investigated methods and provides physiologically meaningful reconstruction results. Our evaluation results provide useful information for choosing cortical source localization approaches for future EEG/MEG studies. PMID- 15784416 TI - A comparison of random field theory and permutation methods for the statistical analysis of MEG data. AB - We describe the use of random field and permutation methods to detect activation in cortically constrained maps of current density computed from MEG data. The methods are applicable to any inverse imaging method that maps event-related MEG to a coregistered cortical surface. These approaches also extend directly to images computed from event-related EEG data. We determine statistical thresholds that control the familywise error rate (FWER) across space or across both space and time. Both random field and permutation methods use the distribution of the maximum statistic under the null hypothesis to find FWER thresholds. The former methods make assumptions on the distribution and smoothness of the data and use approximate analytical solutions, the latter resample the data and rely on empirical distributions. Both methods account for spatial and temporal correlation in the cortical maps. Unlike previous nonparametric work in neuroimaging, we address the problem of nonuniform specificity that can arise without a Gaussianity assumption. We compare and evaluate the methods on simulated data and experimental data from a somatosensory-evoked response study. We find that the random field methods are conservative with or without smoothing, though with a 5 vertex FWHM smoothness, they are close to exact. Our permutation methods demonstrated exact specificity in simulation studies. In real data, the permutation method was not as sensitive as the RF method, although this could be due to violations of the random field theory assumptions. PMID- 15784417 TI - Dynamic shifts in the organization of primary somatosensory cortex induced by bimanual spatial coupling of motor activity. AB - Previous work has shown that training and learning can induce powerful changes in the homuncular organization of the primary somatosensory cortex (SI). Moreover, a number of studies suggest the existence of short-term adaptation of representational maps in SI. Recently, motor activity has been shown to induce rapid modulation of somatosensory cortical maps. It is hypothesized that there is a task-related influence of motor and premotor areas upon the organization of somatosensory cortex. In order to test this hypothesis, we studied the functional organization of somatosensory cortex by examining coupling effects in a bimanual movement task. Bimanual coupling is known to be related to an activation of the premotor cortex and the supplementary motor area. The functional organization of the somatosensory cortex for known bimanual coupling effects was compared to the organization of the somatosensory cortex during the same movements but with only a small effort in coupling. Topography of the functional organization of the somatosensory cortex was assessed using neuromagnetic source imaging based on tactile stimulation of the first (D1) and fifth digit (D5). We could show that the cortical representations of D1 and D5 moved further apart during the bimanual coupling task in comparison to the same task without coupling and rest. Our data suggest that somatosensory cortical maps undergo fast and dynamic modulation as a result of a task-related influence of motor or premotor areas. PMID- 15784418 TI - Processing of famous faces and medial temporal lobe event-related potentials: a depth electrode study. AB - The present study aims at analyzing the modulation of two types of event-related potentials originating from the human medial temporal lobe, the rhinal AMTL-N400 and the hippocampal P600 by the processing of famous faces. Therefore, we used a face recognition paradigm in which subjects had to discriminate the faces of famous persons from the faces of non-famous persons. Eleven patients with unilateral medial temporal lobe epilepsy undergoing intrahippocampal depth electrode recording for presurgical evaluation participated in this study. Event related potentials (ERP) were recorded while a sequence of famous and non-famous faces was presented to the patients. The presentation of each face was repeated. The faces evoked N400-like potentials (anterior medial temporal lobe N400, AMTL N400) in the rhinal cortex and P600-like potentials in the hippocampus. ERPs elicited by famous faces were contrasted with ERPs elicited by non-famous faces. The first presentation of famous faces elicited an enhanced AMTL-N400 and an enhanced hippocampal P600 in comparison to the second presentations of the famous faces or the (first and second presentation of the) non-famous faces. This findings are discussed in terms of associative semantic memory processes and the retrieval of person-specific information from long-term memory stores triggered by the processing of famous faces. PMID- 15784419 TI - The Talairach coordinate of a point in the MNI space: how to interpret it. AB - To perform group studies using functional imaging data, the individual brain images are usually transformed into a common coordinate space. The two most widely used spaces in the neuroscience community are the Talairach space and the Montreal Neurological Institute (MNI) space. The Talairach coordinate system has become the standard reference for reporting the brain locations in scientific publication, even when the data have been spatially transformed into different brain templates (e.g., MNI space). When expressed in terms of individual subjects, the mapping of a coordinate in MNI space to the Talairach space generates distinct coordinates for different subjects. In this paper, we describe two approaches to derive the Talairach coordinates from the MNI space, which is based on the ICBM152 template from the International Consortium of Brain Mapping. One approach is the Talairach Method of Piecewise Linear Scaling (TMPLS) as implemented in the AFNI software package; and the other is a template-matching approach using the linear transformation in SPM99. The uncertainty measurements of the mapping results are presented. This may allow researchers to better interpret results reporting in the Talairach coordinates obtained from the MNI space. This study also examines the discrepancy between the derived Talairach coordinates and those obtained from the mni2tal script, a tool commonly used by the neuroimaging community. Large discrepancies are found in the inferior regions, superior frontal and occipital regions. PMID- 15784420 TI - Opercular to interhemispheric source distribution of benign rolandic spikes of childhood. AB - We evaluated the source distribution of benign rolandic spikes of childhood along and across the central sulcus in 15 patients, aged between 7 and 15 years, who suffered from seizure disorders. Previous routine EEG showed centrotemporal spikes, but none of them had major abnormalities on brain magnetic resonance imaging or neurological deficits. The equivalent current dipoles (ECDs) of the spikes measured by whole-head magnetoencephalography (MEG) were compared to the spike distributions detected by simultaneous scalp EEG according to the international 10-20 system. Locations and orientations of the MEG spikes corresponded to the EEG spike distribution as follows: superiorly oriented spike MEG dipoles in the opercular area corresponded to T3/4 negative peaks (8 spike groups in 6 patients); anteriorly oriented spike dipoles in the rolandic area corresponded to C3/4 or P3/4 negative peaks (17 spike groups in 13 patients); laterally oriented spike dipoles in the interhemispheric area corresponded to Cz/Pz negative peaks (4 spike groups in 3 patients); and others (4 spike groups in 4 patients). Rolandic spikes include three main types according to the ECD location from the opercular to the interhemispheric areas. The functional anatomy of benign rolandic spikes was correlated with partial seizure semiology. All three rolandic spike types can be explained by a precentral origin, assuming that the surface negative potential is continuous from the gyral to fissural cortices. PMID- 15784421 TI - Fiber-tracking does not accurately estimate size of fiber bundle in pathological condition: initial neurosurgical experience using neuronavigation and subcortical white matter stimulation. AB - The fiber-tracking method enables in vivo visualization of the white matter tracts of the brain using a diffusion tensor MR imaging technique. While this method represents a promising tool in the field of neurosurgery, especially when confronted with brain tumors in eloquent areas, its reliability remains unknown. We present here our preliminary validation of tractography in human subjects harboring brain tumors by comparing the results produced by neuronavigation and electrical white matter stimulation in two patients with gliomas in the eloquent area. Although we were able to visualize the pyramidal tract with the fiber tracking technique, the images failed to present the actual size of the fiber bundles. Here we discuss the advantages and limitations of fiber-tracking in the field of neurosurgery. PMID- 15784422 TI - Neurocognitive correlates of incidental verbal memory encoding: a magnetoencephalographic (MEG) study. AB - In our current study, we applied whole head magnetoencephalography (MEG) in a subsequent memory paradigm. Magnetic fields were recorded while 20 healthy subjects (10 females, 10 males) incidentally encoded words during semantic and structural verbal processing tasks. Physiological data were then sorted according to the performance in subsequent memory tests and according to levels of processing, respectively, and analyzed for gender effects. Behavioral results show a clear advantage of semantic processing over structural processing with respect to retrieval success for both females and males. Despite alikeness of behavioral data, MEG results show considerable differences between males and females concerning both subsequent memory effects and levels of processing effects. For male subjects, we identified more distinct magnetic fields in anterior regions for subsequently remembered than for subsequently forgotten words (latency range from 300-650 ms after word onset) and for semantic processing than for structural processing, respectively. For female subjects, corresponding magnetic field differences pointed to posterior regions (subsequent memory effects from 450 ms to 750 ms after word onset). No qualitative differences were observed between subsequent memory effects during semantic processing compared to subsequent memory effects during structural processing. We try to reconcile results from male data with previous findings concerning subsequent memory effects by proposing the concept of width of processing, which holds that incidental memory formation is mediated by frontal activity on a physiological level and brought forward on a cognitive level by enhanced associating imposed by the task demands of semantic processing. Female data cannot be fully incorporated in this framework, but all the more prompt further gender-specific analyses of subsequent memory effects. PMID- 15784423 TI - Separate cortical networks involved in music perception: preliminary functional MRI evidence for modularity of music processing. AB - Music perception is a quite complex cognitive task, involving the perception and integration of various elements including melody, harmony, pitch, rhythm, and timbre. A preliminary functional MRI investigation of music perception was performed, using a simplified passive listening task. Group independent component analysis (ICA) was used to separate out various components involved in music processing, as the hemodynamic responses are not known a priori. Various components consistent with auditory processing, expressive language, syntactic processing, and visual association were found. The results are discussed in light of various hypotheses regarding modularity of music processing and its overlap with language processing. The results suggest that, while some networks overlap with ones used for language processing, music processing may involve its own domain-specific processing subsystems. PMID- 15784424 TI - The effect of encoding strategies on medial temporal lobe activations during the recognition of words: an event-related fMRI study. AB - It is known that manipulation of the encoding strategy affects behavioral and activation data during later retrieval. In the present fMRI study, we examined brain activity during the recognition of words encoded using three different strategies formed by the combination of two factors of relational and self performed processes. The first encoding strategy involved subjects learning words using both relational and self-performed processes (R+S+). In the second, subjects learned words using only a relational process (R+S-). In the third, subjects learned words without using either process (R-S-). During fMRI after encoding, subjects were randomly presented with words encoded previously and with new words (New) and were required to judge whether or not the word presented had been previously encoded. The fMRI experiment was performed with the event-related design. Compared to New, activation of the left medial temporal lobe (MTL) occurred during the recognition of words encoded using R+S+ and R+S-, whereas right MTL activations only occurred with the R+S+ strategy. ROI analysis for the bilateral hippocampus and parahippocampal gyrus showed a linear increase in left MTL activity (hippocampus and parahippocampal gyrus) during the recognition of words encoded with the R-S-, R+S-, to R+S+, whereas right MTL activity (parahippocampal gyrus) was only increased with the R+S+ strategy. The findings suggest that the left and right MTL structures may contribute differentially to the processes involved in the recognition of stimuli and that these differential activities may depend on the encoding strategies formed by the two factors of relational and self-performed processes. PMID- 15784425 TI - Connectivity exploration with structural equation modeling: an fMRI study of bimanual motor coordination. AB - The present fMRI study explores the connectivity among motor areas in a bimanual coordination task using the analysis framework of structural equation modeling (SEM). During bimanual finger tapping at different frequency ratios, temporal correlations of activations between left/right primary motor cortices (MI), left/right PMdc (caudal dorsal premotor area) and supplementary motor cortex (SMA) were detected and used as inputs to the SEM analysis. SEM was extended from its traditional role as a confirmatory analysis to be used as an exploratory technique to determine the most statistically significant connectivity model given a set of cortical areas based on anatomic constraints. The resultant network exhibits coupling from left MI to right MI, links from both PMs to the two MIs, a negative interaction from left PM to right PM, and functional influence from SMA to right MI and right PM, revealing contributions of these areas to bimanual coordination. PMID- 15784426 TI - Combined ICA-LORETA analysis of mismatch negativity. AB - A major challenge for neuroscience is to map accurately the spatiotemporal patterns of activity of the large neuronal populations that are believed to underlie computing in the human brain. To study a specific example, we selected the mismatch negativity (MMN) brain wave (an event-related potential, ERP) because it gives an electrophysiological index of a "primitive intelligence" capable of detecting changes, even abstract ones, in a regular auditory pattern. ERPs have a temporal resolution of milliseconds but appear to result from mixed neuronal contributions whose spatial location is not fully understood. Thus, it is important to separate these sources in space and time. To tackle this problem, a two-step approach was designed combining the independent component analysis (ICA) and low-resolution tomography (LORETA) algorithms. Here we implement this approach to analyze the subsecond spatiotemporal dynamics of MMN cerebral sources using trial-by-trial experimental data. We show evidence that a cerebral computation mechanism underlies MMN. This mechanism is mediated by the orchestrated activity of several spatially distributed brain sources located in the temporal, frontal, and parietal areas, which activate at distinct time intervals and are grouped in six main statistically independent components. PMID- 15784427 TI - Evaluating frequency-wise directed connectivity of BOLD signals applying relative power contribution with the linear multivariate time-series models. AB - In this article, we propose a statistical method to evaluate directed interactions of functional magnetic-resonance imaging (fMRI) data. The multivariate autoregressive (MAR) model was combined with the relative power contribution (RPC) in this analysis. The MAR model was fitted to the data to specify the direction of connections, and the RPC quantifies the strength of connections. As the RPC is computed in the frequency domain, we can evaluate the connectivity for each frequency component. From this, we can establish whether the specified connections represent low- or high-frequency connectivity, which cannot be examined solely using the estimated MAR coefficients. We applied this analysis method to fMRI data obtained during visual motion tasks, confirming previous reports of bottom-up connectivity around the frequency corresponding to the block experimental design. Furthermore, we used the MAR model with exogenous variables (MARX) to extend our understanding of these data, and to show how the input to V1 transfers to higher cortical areas. PMID- 15784428 TI - The many faces of the gamma band response to complex visual stimuli. AB - While much is known about the functional architecture of the visual system, little is known about its large-scale dynamics during perception. This study describes this dynamics with a high spatial, temporal and spectral resolution. We recorded depth EEG of epileptic patients performing a face detection task and found that the stimuli induced strong modulations in the gamma band (40 Hz to 200 Hz) in selective occipital, parietal and temporal sites, in particular the fusiform gyrus, the lateral occipital gyrus and the intra-parietal sulcus. Occipito-temporal sites were the first to be activated, closely followed by the parietal sites, while portions of the primary visual cortex seemed to deactivate temporarily. Some of those effects were found to be correlated across distant sites, suggesting that a coordinated balance between regional gamma activations and deactivations could be involved during visual perception. PMID- 15784429 TI - Neuromagnetic measurement of unilateral temporo-parietal theta rhythm in patients with internal carotid artery occlusive disease. AB - Rhythmic theta activity detected by electroencephalography (EEG) may be correlated with cerebrovascular brain diseases. Magnetoencephalography (MEG) has higher sensitivity and spatial resolution than conventional scalp EEG, so may be a better method to detect theta rhythm in patients with internal carotid artery (ICA) occlusive disease. Simultaneous EEG and MEG were performed in the awake state in 48 patients with unilateral (n = 42) or bilateral (n = 6) stenotic lesions (more than 60% occlusion) of the ICA (n = 47) or middle cerebral artery (n = 7), and in 27 age-matched healthy normal subjects. No subject had severe neurological deficits. MEG detected the theta rhythm (6-8 Hz) in 14 of 48 patients: ipsilateral to the stenotic or occluded side in 13 hemispheres and bilaterally in one patient with unilateral lesion. The source of the MEG theta rhythm was estimated in the dorsolateral temporo-parietal area, regardless of the location of infarct foci or the stenotic portion of the ICA system. The temporo parietal theta rhythm was separated from the occipital alpha rhythm by frequency and distribution in MEG. The theta rhythm was found in only two patients by EEG, as well as by MEG. MEG provided better separation of this theta rhythm from the occipital alpha rhythm. Neither MEG nor EEG detected this theta rhythm in the normal subjects. Unilateral temporo-parietal theta rhythm is correlated with the hemisphere with ICA occlusive disease. This rhythm may indicate mild or subclinical abnormalities in the ICA system. MEG is superior to EEG for the detection and localization of theta rhythm. PMID- 15784430 TI - Task demands modulate sustained and transient neural activity during visual matching tasks. AB - The extent to which a task demands verbal or non-verbal processing may influence which neural regions underlie performance. In the present study, sustained and transient responses were examined using functional magnetic resonance imaging (fMRI) in conjunction with a mixed blocked/event-related design during three visual-matching tasks that varied in the extent to which they relied on verbal processing. In a name-matching task, subjects decided whether two letters had the same or a different name (e.g., A a); in a physical-matching task, subjects decided whether two letters were exactly the same or different (e.g., A A); in a non-letter-matching task, subjects decided whether two non-letters were exactly the same or different. Results revealed several regions in which sustained activity differed across the three tasks as well as several regions in which sustained activity did not differ. Most notably, regions in the right inferior frontal gyrus exhibited greater sustained activity during the name-matching task than during the physical or non-letter-matching tasks, indicating that sustained activity in this region is sensitive to the amount of verbal processing required by a particular task. Moreover, transient activity in the right inferior frontal regions, as well as others, exhibited the opposite pattern of results. In combination, results suggest that sustained and transient activities interact to produce the context-appropriate response during visual-matching tasks. PMID- 15784431 TI - Neuroimaging of emotion: empirical effects of proportional global signal scaling in fMRI data analysis. AB - Global variations of BOLD-fMRI signal are often considered as nuisance effects. This unwanted source of variance is commonly eliminated using proportional global signal scaling (PGSS). However, application of PGSS relies on the assumption that global variations of BOLD signal and experimental conditions are uncorrelated. It has been shown for cognitive tasks that the unjustified application of PGSS might greatly distort statistical results. The present study examined this issue in the domain of emotion research. Specifically, fMRI data were obtained in a block design, while 21 subjects passively viewed high and low emotionally arousing pleasant, unpleasant, and neutral pictures. Violations of the orthogonality assumption were found for analyses of emotional pictures high in arousal, causing dramatically different outcomes when compared to analyses performed without PGSS. Application of PGSS was associated with attenuated emotional activation in visual cortical areas, insensitivity to emotional activations in limbic and paralimbic regions, and widely distributed artificial deactivations. In contrast, the orthogonality assumption was not violated for low arousing emotional materials. Thus, the validity of using PGSS varied as a function of the emotional arousal of the stimuli. Taken together, the unwarranted use of PGSS might contribute to conflicting results in affective neuroscience fMRI studies, in particular with respect to limbic and paralimbic structures. PMID- 15784432 TI - Semi-blind ICA of fMRI: A method for utilizing hypothesis-derived time courses in a spatial ICA analysis. AB - Independent component analysis (ICA) is a data-driven approach utilizing high order statistical moments to find maximally independent sources that has found fruitful application in functional magnetic resonance imaging (fMRI). Being a blind source separation technique, ICA does not require any explicit constraints upon the fMRI time courses. However, for some fMRI data analysis applications, such as for the analysis of an event-related paradigm, it would be useful to flexibly incorporate paradigm information into the ICA analysis. In this paper, we present an approach for constrained or semi-blind ICA (sbICA) analysis of event-related fMRI data by imposing regularization on certain estimated time courses using the paradigm information. We demonstrate the performance of our approach using both simulations and fMRI data from a three-stimulus auditory oddball paradigm. Simulation results suggest that (1) a regression approach slightly outperforms ICA when prior information is accurate and ICA outperforms the general linear model (GLM)-based approach when prior information is not completely accurate, (2) prior information improves the robustness of ICA in the presence of noise, and (3) ICA analysis using prior information with temporal constraints can outperform a regression approach when the prior information is not completely accurate. Using fMRI data, we compare a regression-based conjunction analysis of target and novel stimuli, both of which elicit an orienting response, to an sbICA approach utilizing both the target and novel stimuli to constrain the ICA time courses. Results show similar positive associations for both GLM and sbICA, but sbICA detects additional negative associates consistent with regions implicated in a default mode of brain activity. This suggests that task-related default mode decreases have a more "complex" signal that benefits from a flexible modeling approach. Compared with a traditional GLM approach, the sbICA approach provides a flexible way to analyze fMRI data that reduces the assumptions placed upon the hemodynamic response of the brain. The advantages and limitations of our technique are discussed in detail in the manuscript to provide guidelines to the reader for developing useful applications. The use of prior time course information in a spatial ICA analysis, which combines elements of both a regression approach and a blind ICA approach, may prove to be a useful tool for fMRI analysis. PMID- 15784433 TI - Validation of SOBI components from high-density EEG. AB - Second-order blind identification (SOBI) is a blind source separation (BSS) algorithm that can be used to decompose mixtures of signals into a set of components or putative recovered sources. Previously, SOBI, as well as other BSS algorithms, has been applied to magnetoencephalography (MEG) and electroencephalography (EEG) data. These BSS algorithms have been shown to recover components that appear to be physiologically and neuroanatomically interpretable. While some proponents of these algorithms suggest that fundamental discoveries about the human brain might be made through the application of these techniques, validation of BSS components has not yet received sufficient attention. Here we present two experiments for validating SOBI-recovered components. The first takes advantage of the fact that noise sources associated with individual sensors can be objectively validated independently from the SOBI process. The second utilizes the fact that the time course and location of primary somatosensory (SI) cortex activation by median nerve stimulation have been extensively characterized using converging imaging methods. In this paper, using both known noise sources and highly constrained and well-characterized neuronal sources, we provide validation for SOBI decomposition of high-density EEG data. We show that SOBI is able to (1) recover known noise sources that were either spontaneously occurring or artificially induced; (2) recover neuronal sources activated by median nerve stimulation that were spatially and temporally consistent with estimates obtained from previous EEG, MEG, and fMRI studies; (3) improve the signal-to-noise ratio (SNR) of somatosensory-evoked potentials (SEPs); and (4) reduce the level of subjectivity involved in the source localization process. PMID- 15784434 TI - Altered patterns of blood flow response during visual stimulation in carotid artery occlusive disease. AB - To correctly interpret the results of functional neuroimaging studies in stroke, it is essential to understand how cerebrovascular disease influences hemodynamic response during neural activation. To investigate the effect of internal carotid artery (ICA) occlusive disease on the pattern of cerebral blood flow (CBF) response during visual stimulation, we used positron emission tomography to study 13 patients with ICA steno-occlusive lesions. We measured the changes of CBF during visual stimulation in the primary visual cortex and in the surrounding region, including the higher-order visual cortex, and examined their correlation with the baseline value of oxygen extraction fraction, a measure of hemodynamic impairment, in the ICA distribution. With visual stimulation, CBF in the primary visual cortex significantly increased in all patients, while in the surrounding region, CBF showed variable changes, including decreases in some patients. In 9 patients with unilateral ICA lesions, the CBF change in the surrounding region ipsilateral to the ICA lesion was significantly decreased compared with the value in the contralateral hemisphere, while the CBF change in the primary visual cortex showed no hemispheric difference. The hemispheric values of oxygen extraction fraction in the ICA distribution and the amount of CBF increase in the visual cortex were independently and negatively correlated with the CBF change in the surrounding region. We conclude that the pattern of CBF response during visual stimulation may change in ICA occlusive disease. We suggest that the redistribution of CBF during visual stimulation may be a contributing mechanism. PMID- 15784435 TI - Hippocampal functional magnetic resonance imaging during a face-name learning task in adolescents with antecedents of prematurity. AB - Functional magnetic resonance imaging (fMRI) was used to map hippocampal activation during a declarative memory task in a sample of 14 adolescents with antecedents of prematurity (AP). The sample with AP was matched by age, sex and handedness with 14 full-term controls with no history of neurological or psychiatric illness. The target task consisted in learning 16 novel face-name pairs, and the control task involved the examination of two repeated face-name pairs. Stereological methods were also used to quantify hippocampal volumes. In both groups, we observed increased activation in the learning condition compared to the control task in the right fusiform gyrus and the left inferior occipital gyrus, but only premature subjects activated the hippocampus. Group comparison of the activation versus control conditions showed that prematures had greater activity in the right hippocampus than controls during the encoding of the word face association. Volumetric analyses showed a significant left hippocampal volume loss in adolescents with AP. In addition, we found a significant positive correlation in the premature group between right hippocampal activation and face name recognition. Functional MRI data also correlated with structural MRI data: right hippocampal activation correlated positively with right hippocampal volume. Our findings are consistent with previous studies of brain plasticity after focal lesions. Left hippocampal tissue loss may be related to an increase in contralateral brain activity, probably reflecting a compensatory mechanism. Our data also suggest that this plasticity is not enough to achieve normal performance. PMID- 15784436 TI - The appreciation of wine by sommeliers: a functional magnetic resonance study of sensory integration. AB - We set out to investigate how the expertise of a sommelier is embodied in neural circuitry by comparing brain activity elicited by wine tasting with that found in naive drinkers of wine. We used fMRI to study 7 sommeliers and 7 age- and sex matched control subjects to test the hypothesis that any difference in brain activity would reflect a learned ability to integrate information from gustatory and olfactory senses with past experience. A group analysis showed activation of a cerebral network involving the left insula and adjoining orbito-frontal cortex in sommeliers. Both these areas have been implicated in gustatory/olfactory integration in primates. In addition, activation was found bilaterally in the dorsolateral prefrontal cortex, which is implicated in high-level cognitive processes such as working memory and selection of behavioral strategies. Naive individuals activated the primary gustatory cortex and brain areas, including the amygdala, implicated in emotional processing. PMID- 15784437 TI - Dissociation of cortical regions modulated by both working memory load and sleep deprivation and by sleep deprivation alone. AB - Working memory is an important mental capacity that is compromised following sleep deprivation (SD). To understand how working memory load interacts with state to influence brain activation in load-sensitive regions, and the extent to which SD-related changes are common across different loads, we used fMRI to study twelve healthy subjects following 24 h of SD using a verbal n-back task with three load levels. Performance decline was observed by way of reduced accuracy and slower response times following SD. The left prefrontal region and thalamus showed load dependent activity modulation that interacted with state. The right parietal and anterior medial frontal regions showed load dependent changes in activity as well as an effect of state. The anterior cingulate and occipital regions showed activation that displayed state effects that were independent of working memory load. These findings represent a step toward identifying how different brain regions exhibit varying vulnerability to the deleterious effects of SD on working memory. PMID- 15784438 TI - Executive dysfunction in Parkinson's disease is associated with altered pallidal frontal processing. AB - Executive dysfunction in Parkinson's disease is well documented, but it is still unclear whether this results from (i) prefrontal dysfunction, (ii) striatal dysfunction, or (iii) altered striatal outflow to the prefrontal cortex. To clarify this issue, we used H(2)(15)O PET to asses six nondemented and nondepressed patients with Parkinson's disease and six matched controls while they performed a task involving executive function, random number generation (RNG), and a control counting task. To assess the effect of increasing task demands, each task was performed at three rates. Both groups showed significant increase in nonrandomness of responses during RNG at faster rates, which was differentially greater for the patients at the faster rate. The controls showed significant activation of the lateral and medial prefrontal cortex and superior and medial parietal cortex during RNG relative to counting. For the same comparison, the patients did not show any activity in medial frontal structures. The controls showed significantly greater mesial frontotemporal activation during counting than RNG, whereas the patients did not show any modulation of regional cerebral blood flow (rCBF) in these areas with task. With faster rates of RNG, the controls showed rCBF increase in the right internal segment of globus pallidus (GPi) and a decrease in frontal cortex. The patients showed the opposite pattern of subcortical and frontal rCBF change with faster rates. The results suggest that executive dysfunction in Parkinson's disease is associated with a failure to modulate frontal activation with increased task demands (nature of task or rate), a deficit associated with altered rCBF in the GPi, the final basal ganglia output pathway to frontal cortex rather than any intrinsic prefrontal dysfunction. PMID- 15784439 TI - Dissociation of neural systems mediating shifts in behavioral response and cognitive set. AB - The ability to generate appropriate behaviors in response to changing situations requires both the alteration of ongoing behavior and the understanding of the global rules governing stimulus categorization in a given context. Neuropsychological tests that have been developed to measure this form of cognitive flexibility, such as the Wisconsin Card Sorting Test, have reliably demonstrated that individuals with lesions in regions of the prefrontal cortex and basal ganglia have difficulty generating a cognitive set and altering rule governed behavior. Recent neuroimaging studies have supported the role of these brain regions in the performance of response shifting and cognitive set shifting. However, the precise role of these regions in the individual components of these tasks remains a contentious issue. Here, we used event-related functional magnetic resonance imaging (fMRI) to dissociate the neural circuitry involved in the alteration of ongoing behavior and the shifting of cognitive set. Participants viewed geometric shapes as they appeared individually in rapid succession and responded with an appropriate button press based upon whether the individual shape was a predetermined target stimulus. Responses were required for each shape presented. The fMRI results indicated that response shifting specifically activated a dorsal neural circuit comprised of the dorsolateral prefrontal cortex, anterior cingulate, and intraparietal sulcus. Shifts in cognitive set were mediated by ventrolateral prefrontal cortex, anterior cingulate, and striatum. These findings suggest that the alteration of ongoing behavior and shifting of cognitive set are mediated by two distinct neural systems interconnected by the anterior cingulate. PMID- 15784440 TI - Superior temporal gyrus and insula provide response and outcome-dependent information during assessment and action selection in a decision-making situation. AB - Decision-making is a complex process that comprises the assessment of a situation, the selection of an action, and the evaluation of an outcome. Distinct neural systems may contribute differentially during various stages within a decision-making situation. This study investigated whether neural activation during assessment or action selection is critically dependent on previous outcomes or actions. Twelve healthy, right-handed subjects (6 females) played a Rock Paper Scissors (RPS) computer game during functional magnetic resonance imaging. Bilateral insula and medial prefrontal cortex (including the anterior cingulate) were specifically engaged during the assessment and action selection stages of decision-making, whereas bilateral superior frontal gyrus and right inferior parietal lobule activated more during the outcome. Two regions of activation within the bilateral superior temporal gyrus activated only when the previous outcome was a win. Moreover, right insula and superior temporal gyrus were active more when the subject switched responses relative to staying with the same choice made on the previous trial. These findings support the hypothesis that distinct neural systems underlie different stages of the decision-making process. Furthermore, the superior temporal gyrus may play an important role in integrating previous actions and successful outcomes into one's decision-making strategy. PMID- 15784441 TI - Self-referential reflective activity and its relationship with rest: a PET study. AB - This study used positron emission tomography (PET) to identify the brain substrate of self-referential reflective activity and to investigate its relationship with brain areas that are active during the resting state. Thirteen healthy volunteers performed reflective tasks pertaining to three different matters (the self, another person, and social issues) while they were scanned. Rest scans were also acquired, in which subjects were asked to simply relax and not think in a systematic way. The mental activity experienced during each scan was assessed with rating scales. The results showed that, although self referential thoughts were most frequent during the self-referential task, some self-referential reflective activity also occurred during rest. Compared to rest, performing the reflective tasks was associated with increased blood flow in the dorsomedial prefrontal cortex, the left anterior middle temporal gyrus, the temporal pole bilaterally, and the right cerebellum; there was a decrease of blood flow in right prefrontal regions and in medial and right lateral parietal regions. In addition, the ventromedial prefrontal cortex (VMPFC) (1) was more active during the self-referential reflective task than during the other two reflective tasks, (2) showed common activation during rest and the self referential task, and (3) showed a correlation between cerebral metabolism and the amount of self-referential processing. It is suggested that the VMPFC is crucial for representing knowledge pertaining to the self and that this is an important function of the resting state. PMID- 15784442 TI - Sensitivity-encoded (SENSE) echo planar fMRI at 3T in the medial temporal lobe. AB - Parallel imaging techniques are useful for fMRI studies in light of the increasing susceptibility effects at high magnetic field strength. Yet, spatially varying noise amplification constitutes a challenge for the application of these techniques. The medial temporal lobe is particularly vulnerable to susceptibility effect with increasingly strong signal reduction. We present two fMRI studies comparing SENSE single-shot (ssh) echo planar imaging (EPI) at acceleration factors of 2.0, 2.4, 2.7, and 3.0 with conventional sshEPI at TE of 22 and 35 ms. Data were acquired during a learning task which activates the medial temporal lobe bilaterally. Susceptibility related image distortion was markedly reduced with increasing SENSE acceleration. Moreover, in the group results, statistical power increased in the whole brain with SENSE compared to conventional imaging and with a TE of 35 ms compared to 22 ms. Higher SENSE acceleration factors further improved image quality and increased statistical power in the occipital lobe and fusiform gyrus, but not in the medial temporal lobe. We therefore conclude that an sshEPI acquisition protocol with a moderate SENSE acceleration factor of R = 2.0 and TE 35 ms is suitable for the detection of medial temporal activation at 3T. PMID- 15784443 TI - Visualization of temporal increase in compound nerve action magnetic fields in the human median nerve during ischemia. AB - We studied the variation in the human median nerve activity during ischemia by measuring compound nerve action magnetic fields (CAFs). Temporal increases in the CAF during ischemia were successfully visualized on isofield contour maps. Intense paresthesias were induced after 29 +/- 5 s (mean +/- SD) of ischemia, which consistently reached a maximum after 4 min 55 +/- 12 s. The variations in CAFs were consistent with changes in sensory perception. The hyperexcitability in large myelinated axons can be visualized as temporary increases in CAF. These results are the first to demonstrate the efficacy of magnetic-recording techniques, which allow monitoring of changes in neural activity. PMID- 15784444 TI - Magnetic resonance imaging of the human frontal cortex reveals differential anterior-posterior variability of sulcal basins. AB - MRI data of 100 healthy human brains were analyzed to establish a neuroanatomical map of the most frequently occurring 'sulcal basins' of the human frontal cortex. Sulcal basins are defined to be concavities in the white matter surface constituting/representing components of entire sulci. We determined their volume, depth, and interindividual variability. The sulcal basins were found to fall into two groups, on average, eight anterior basins in the prefrontal and premotor region and four posterior ones in the motor region of the frontal lobe. Compared to posterior basins, anterior basins are characterized by lower volume and depth. Furthermore, they showed greater interindividual variability in volume, depth, and occurrence. Our results indicate the existence of a mechanism for cortical folding which shows a greater flexibility in the phylogenetically younger, anterior prefrontal areas. PMID- 15784446 TI - Malaria problem in Afghanistan: malaria scanning results of the Turkish medical aid group after the war. AB - Malaria is a parasitic infection caused by Plasmodium species and it is especially seen in tropical and subtropical areas. We aimed to evaluate the effects of the infection in Afghanistan, which is an endemic place for malaria and had severe socio-economical lost after the war. We also compared these data with the ones that were recorded before the war. Blood samples were taken from 376 malaria suspected patients who come to the health center, established by the medical group of Istanbul Medical Faculty in 2002, Afghanistan. Blood samples were screened using the OPTIMAL Rapid Malaria Test and Giemsa staining method. In 95 (25.3%) patients diagnosis was malaria. In 65 patients (17.3%) the agent of the infection was P. falciparum and in 30 patients (8%) agents were other Plasmodium species. PMID- 15784448 TI - Advances in haemapheresis and haemotherapy. PMID- 15784447 TI - Circulation and distribution of 111-In-oxine-labeled autologous baboon platelet aggregates and buffy coat. AB - BACKGROUND: Although it is known that RBC concentrates may contain buffy coat and platelet concentrates may contain platelet aggregates, the circulation and distribution of these materials in the blood products have never been reported. STUDY DESIGN AND METHODS: Baboon platelets were labeled with 111-In-oxine, aggregated with ADP and autotransfused without a filter. Baboon buffy coat was stored at 4 degrees C, labeled with 111-In-oxine and autotransfused without a filter. The circulation of the radiolabeled platelets and buffy coat was measured and the distribution of the buffy coat and platelet aggregates was measured by external scanning of the baboon using a gamma camera. The effects of the infusion of aggregated platelets, buffy coat, and gelatin on the plasma fibronectin level also were evaluated. RESULTS: The 111-In-oxine labeled platelet aggregates were initially sequestered in the lungs and released into the peripheral blood during the next 3h, during which time the cell associated radioactivity increased by about 25%. Following the autotransfusion of 111-In-oxine labeled buffy coat, the 111-In-oxine radioactivity over the lungs increased, but decreased during the 60 min post-transfusion period as the radioactivity over the liver increased. Cell associated radioactivity increased by about 10% over the 3-h post-transfusion period. Fibronectin levels decreased by 3% following the autotransfusion of platelet aggregates, by 10% after the autotransfusion of buffy coat and by 50% after the infusion of gelatin. CONCLUSIONS: 111-In-oxine radioactivity in the platelet aggregates and buffy coat was initially sequestered in the lungs, and 10 25% of the 111-In-oxine cell-associated radioactivity was released into the circulation during the 24-h post-transfusion period. PMID- 15784449 TI - Optimization of the therapeutic procedure during LDL-apheresis--a computerized model. AB - LDL-apheresis is a very effective method for the treatment of resistant hypercholesterolemia when other therapy (dietary, or medication) fails. To maximize the efficacy of the LDL-absorbers we aimed to create a computerized model. PATIENTS AND METHODS: The therapeutic technique of immunoadsorption was used, applying a pair of columns, the Lipopak, Pocard, Russia. Plasma was separated by a continuous-flow plasma separator, the Cobe Spectra, USA; adsorption was controlled by adsorption-desorption equipment, ADA, Medicap, Germany. 494 LDL-apheresis procedures (treatment interval 17.5+/-1.6 days) were used to treat nine patients with primary hypercholesterolemia followed during the consecutive 3.6+/-0.5 years. Metabolism of LDL-cholesterol is known to be multicompartmental and dynamic, but for the short-time period of the procedure it can be simplified and one can calculate the procedure as a continuous filtration. We developed a program for procedure planning, using Microsoft Excel for Windows. Inputs inserted into the program include only basic patient data (mass, height, sex and initial plasma LDL level in mmol/l). RESULTS: The results show a very promising match between our planning of the procedures and the real laboratory results. The drop in calculated vs real plasma LDL-cholesterol level differ no more than +/-10%. CONCLUSIONS: Although our software does not take into account many well known details about the metabolism of cholesterol, in given conditions it can provide a fairly precise prediction of procedure parameters. It is also suitable for practical use, because it requires only a few commonly used and readily available input values. Immunoadsorption with Pocard absorbers is a potent and safe method of therapy in indicated patients. PMID- 15784450 TI - The transplantation of neural stem cells and predictive factors in hematopoietic recovery in irradiated mice. AB - A number of surprising observations have shown that stem cells, in suitable conditions, have the ability to produce a whole spectrum of cell types, regardless, whether these tissues are derived from the same germ layer or not. This phenomenon is called stem cell plasticity, which means that tissue-specific stem cells are mutually interchangeable. In our experiments, as a model, we used neural stem cells (NSCs) harvested from fetal (E14-15) neocortex and beta galactosidase positive. In the first experiment we found that on days 12 and 30 after sub-lethal irradiation (LD 8.5 Gy) and (beta-galactosidase(+)) NSCs transplantation all mice survived, just as the group with bone marrow transplantation. Moreover, the bone marrow of mice transplanted NSCs contained the number of CFU-GM colonies with beta-galactosidase(+) cells which was as much as 50% higher. These differences were statistically significant, p<0.001. In the second experiment, we studied kinetics of (beta-galactosidase(+)) NSCs after their transplantation to sub-lethally irradiated mice. Histochemistry of tissues was performed on days 12 and 30 post-transplantation, and beta-galactosidase(+) cells were detected with the help of histochemical examination of removed tissues (lung, liver, spleen, thymus, and skeletal muscle). In tissues removed on day 12 post-transplantation, we found a significantly higher number of beta galactosidase(+) cells in the spleen and thymus on day 30. While we presumed the presence beta-galactosidase(+) cells in the spleen, as spleen and reticuloendothelial system represent an important retaining system for different cell types, the presence of beta-galactosidase(+) cells in the thymus was rather surprising but very interesting. This indicates a certain mutual and close interconnection of transplanted stem cells and immune system in an adult organism. In the third experiment, we verified the mutual interchange of Sca-1 surface antigen in the bone marrow cells and NSCs before transplantation. Analysis of this antigen showed 24.8% Sca-1 positive cells among the bone marrow cells, while NSCs were Sca-1 negative. Our experiments show that NSCs share hemopoietic identity and may significantly influence the recovery of damaged hematopoiesis but do not have typical superficial markers as HSCs. This result is important for the determination of predictive factors for hemopoiesis recovery, for stem cell plasticity and for their use in the cell therapy. PMID- 15784451 TI - PBPC collection techniques: standard versus large volume leukapheresis (LVL) in donors and in patients. AB - Transplantations of autologous and allogeneic peripheral blood progenitor cells (PBPC) are able to assure a complete hematopoietic and immunologic reconstitution in patients. PBPC are collected by leukapheresis technique after prior mobilization therapy, but procedures and results remain still highly variable and are poorly characterized. An optimum regimen for PBPC collections has not yet been recommended, but 2-3 total blood volumes (TBV) of the donor or patient are regarded as a standard. Another promising technique is large volume leukapheresis (LVL) with processing of 3-6 TBV of donor or patient. The aim of this paper is to find the most efficient and safe collection technique for an individual donor or patient and, consequently minimize the number of procedures required. Finding the optimal collection procedure would be helpful while considering which method would be preferred in an individual donor or patient with respect to the result of mobilization, health state and required yield of CD 34+ cells for transplantation. We evaluated the results in a total of 134 standard and LVL procedures, which were performed in 21 well mobilized donors (Group I), in 65 well mobilized patients (Group II), and in 14 weakly mobilized patients (Group III) with hemato-oncological diseases. A precollection concentration of CD 34+ cells in peripheral blood higher than 20 x 10(3)/mL was considered to be the criterion for efficient mobilization. Such levels of concentration indicating the start of PBPC collections could be easily reached in Group I of donors and Group II of well mobilized patients. Heavily pretreated patients at advanced stages of disease (Group III) did not respond to mobilization sufficiently and had a concentration of CD 34+ cells lower than 20x10(3)/mL. LVL technique made it possible to obtain higher numbers of CD 34+ cells than in the standard collection in well mobilized donors (Group I), well mobilized patients (Group II), and even in weakly mobilized patients in Group III. In donors and well mobilized patients (Group I and Group II) it was possible to collect sufficient amounts of CD 34+ cells for allogeneic or for autologous transplantation from one LVL collection. The median yield of CD 34+ cells from one LVL collection was 5.5 x 10(6)/kg b.w. in donors, and 6.0 x 10(6)/kg b.w. in well mobilized patients. Due to the linear dependence of the yield of collected CD 34+ cells on the concentration of CD 34+ cells in blood, it can be used as a simple prediction of the success of collection in Group II (correlation coefficient 0.93 for standard procedures, and correlation coefficient 0.88 for LVL). In Group III of weakly mobilized patients the standard collections were usually ineffective and the relationship between the yield of CD 34+ cells/kg in the product and the precollection concentration of CD 34+ cells was much less significant (correlation coefficient 0.56 for standard procedures and correlation coefficient 0.66 for LVL). The median of CD 34+ cells collected from one standard procedure was only 0.7 x 10(6)/kg but using LVL the median increased to 1.4 x 10(6)/kg. Our results prove that the yield of CD 34+ cells in the product can be enhanced by large volume leukapheresis (LVL). Based on the results obtained, we recommend LVL in all donors and patients who can tolerate it due to a greater chance of collecting higher yields of progenitor cells while minimizing adverse reactions. LVL procedures should also be preferred in weakly mobilized patients where it is not possible to collect sufficient amounts of CD 34+ cells for transplantation using the standard regime. In weakly mobilized patients LVL provides a greater chance to at least collect a minimum amount of CD 34+ cells necessary. LVL should be used in circumstances where extremely high doses of CD 34+ cells has to be prepared, e.g. planned "tandem" transplantations or manipulations with a graft in which a significant loss of cells is expected. PMID- 15784452 TI - The "cutting" edge: von Willebrand factor-cleaving protease activity in thrombotic microangiopathies. AB - The thrombotic microangiopathies (TMAs), including thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS), are classically defined by constellations of clinical findings. In the late 1990's, several groups reported that a single test might be able to diagnose TTP and distinguish it from HUS. This test was an assay to detect von Willebrand factor-cleaving protease activity. Although there has been debate in the literature as to the utility of this assay, review of the available data suggests that this test can separate the idiopathic TMAs into three distinct categories: (1) patients who are protease positive with severe renal failure (similar to classic HUS), (2) patients who are protease positive without severe renal failure and (3) patients who are protease negative. While the latter two categories may appear similar to TTP at presentation, the prognosis differs in each of these groups of patients. Through use of the protease assay clinicians will be better able to diagnose, treat, predict outcome and design clinical trials. PMID- 15784453 TI - Modern concepts of autologous haemotherapy. AB - The increasing current perception that the safety of allogeneic blood transfusion has dramatically been improved during the last decade is challenging autologous haemotherapy methods. In addition, growing concern about the unfavourable cost effectiveness of most autologous haemotherapy methods requires a refinement of the application of these measures to well defined circumstances. In contrast, newly emerging transfusion-transmissible infections or periods of blood shortage might revive interest in these blood sparing techniques. Preoperative autologous blood donation still plays a significant role in settings with high individual benefit for the patient, high transfusion probabilities and when all opportunities of cost minimization can be applied. Preoperative plasmapheresis is considered to be a sensible adjunct if intraoperative retransfusion of salvaged and washed red cells is planned. Acute normovolaemic haemodilution is valuable when the patient's tolerability of the haemodilution and the expected blood loss are carefully examined beforehand. Intra- or postoperative salvage of wound blood can also be regarded as useful measures to prevent allogeneic transfusions as long as the specific advantages and disadvantages of the different methods are taken into account. Finally, alternative and supplemental measures such as iron or erythropoietin administration should always be considered in order to optimize the efficacy and effectiveness of autologous haemotherapy methods. The goal of a "bloodless medicine" might not be reached but is supposed to be approached closely with an integrated concept exploiting all measures available. However, in times of restricted health care resources, regular sound cost-effectiveness analyses, taking the availability and the current safety profile of allogeneic blood products into account, are always warranted and needed. PMID- 15784454 TI - Small-scale extracorporeal photopheresis for the treatment of cutaneous T-cell lymphoma: a report of 3 cases. AB - Extracorporeal photopheresis is an accepted method for the treatment of cutaneous T-cell lymphoma and much progress has recently been achieved in therapy and understanding of its mechanism. In general large numbers of white blood cells are collected by a cell separator and irradiated in the presence of 8-MOP. In contrast to this practice, data from an animal model showed that as few as 0.2% of the body's blood volume irradiated are sufficient to achieve an immune response after photopheresis. Based on these data we developed a small-scale photopheresis procedure and applied the method in 3 end-stage T-cell lymphoma patients who were not eligible for apheresis. The mononuclear cells from 50 ml of blood were separated by density gradient centrifugation, irradiated with UV-light in the presence of 8-Methoxy-Psoralen (MOP) with 2J/cm(2) and reinjected. 2-3 treatments per week were conducted. The three patients-2 male and 1 female, age 63-86, Sezary syndrome (1x) and mycosis fungoides in tumour stage (2x)-showed no side effects on cell injection. The two patients with mycosis fungoides showed a prompt regression and softening of the tumours. The patient with Sezary syndrome developed numerous necrotic spots on the skin after 6 weeks of therapy that turned normal within a few days. Patient 1 died of pneumonia 4 weeks after the start of therapy and patient 3 died of heart failure 8 weeks after start of therapy, both during regression of the tumours. Patient 2 was treated over a period of 11 months, with an initial regression in the first weeks followed by a slow progression of the tumours after she rejected any form of further treatment. The small-scale extracorporeal photopheresis therapy presented is effective in cutaneous T-cell lymphoma. But questions regarding the optimal number of cells irradiated per treatment, the conditions of cell incubation after irradiation and the number of treatment cycles are still open. Therefore further studies are required to establish a method that is effective and circumvents the use of apheresis technology. PMID- 15784455 TI - World apheresis association--world apheresis registry. AB - In 2002 WAA decided to start a world-wide apheresis registry to gain insight into the extent of treatment, adverse events, and to facilitate contacts among centers when treatment indications are rare and experience limited. Stem cell and other blood products collections intended for therapeutic application can also be entered. The WAA planned to use the French Registry. Its translation into English has not been accomplished and the fiscal obligations for that registry has not, as yet, been determined or considered and approved by the WAA Board. From Dec 2002 the proposed registry (a merged version of the French, Canadian and Swedish registries) can be immediately implemented. We now cordially invite all centers to join that registry. Please, also inform colleagues at other centers in your country to join. E-mail and address lists of colleagues in your country who have not registered will be welcomed. The site is at: Go to World Apheresis Registry; Login code to test the Registry is: al61tms. Then apply for a specific login code for your center. We welcome you to this registry for your input of data. You will not be charged any registration fee. The registry includes a randomization system that can be used for local or multi center studies (randomization by in-center basis allows you to make your own studies). It includes a formula that increases the chance to get a more even distribution between groups also for smaller sample sizes. PMID- 15784456 TI - A survey of blood purification techniques. AB - Apheresis may be performed with many different techniques. The basis for different therapeutic approaches lies in the pathophysiological processes present in the diseases that have to be treated. Over the years more sophisticated devices have been developed. The most frequent treatment is plasma exchange (plasmapheresis) using centrifugation or single filtration techniques. In addition cascade filtration and subsequent adsorption from plasma is done. Thereby removal is done by adsorption of molecules such as bilirubin, immunoglobulins (immunoadsorption), circulating immune complexes, various antibodies including those against blood types. Such adsorption technologies have also been developed to allow adsorption directly from a column perfused by whole blood (hemoperfusion). By combining various techniques, systems are available that allow bridging of patients with hepatic failure to transplantation (MARS, Prometheus). By adding e.g., hepatic cells to such systems, besides dialysis and adsorption, cells will help to degrade toxic molecules. Such bioreactors are in clinical use. Apheresis includes also the removal or retrieval of cells from blood for e.g., stemcell transplantation, polycythaemia or hemochromatosis. Removal of leukocytes from blood using leukocyte filters is indicated in inflammatory bowel diseases. By specifically irradiating lymphocytes and monocytes with UV light using the technique of extra corporeal photochemotherapy (ECTP) various immunological diseases are treated. On the other hand, various alternative techniques may be used for the same disorder. Thus for patients with high plasma LDL-cholesterol not responding to other lipid lowering strategic treatment, alternative therapy may be done either by cascade filtration, adsorption technology from plasma, heparin precipitation (HELP-system) or hemoperfusion. This article describes various techniques in clinical use. PMID- 15784457 TI - The content of the LRS chamber provides a new quality tool for the characterization of the donor platelet profile. AB - Cobe Trima version 4 is an apheresis system designed for the collection of combinations of Red Blood Cells (RBC), Platelets (PLT) and plasma components from a single donor. The validation of this apheresis system for PLT components in our institution evidenced the LRS efficacy, as leucoreduction was attained on a 100% basis. However, there were some unexpected occurrences regarding the PLT content. Certain donations showed large disagreements between programmed and obtained yields, not being found clear reasons for those outcomes. Furthermore, the mean platelet volume (MPV) was relatively low when compared with other platelets components produced by other methodologies. An investigation was initiated in order to know whether these shortcomings were donor or process/instrument related. A link was established between the raw material (donor), the process/instrument and the final product, and a new tool was introduced by the study of the LRS chamber. The LRS chamber content was assessed and the PLT cellular indices and PLT aggregation states compared with those obtained from the respective donor and the final product from the same origin. The storage stability of the final products was also analyzed based on these same tests to investigate if the initial low MPV had any deleterious effect during the shelf life of the components. Twenty five plateletpheresis donors, three of them new ones, were randomly selected for this study. For the first time, the content of the LRS chamber was used as a quality tool for identification of the cause of unexpected yields. Large aggregates remained in the LRS chamber in certain donors who were prone to undergo spontaneous aggregation, making the platelet yields low and platelet MPV very small. Based on repeated failures and on this new quality parameter showing LRS chamber abnormality, two donors were temporarily deferred from the panel. While it was not possible to identify the causes, it is appropriate to raise the question whether these profiles were Trima version 4 specific or not. Hence, further investigation is needed for a better clarification. In short, the simultaneous analysis of products and LRS content provided useful information not only for the characterization of donor-related phenomena but also helped in the identification of potential shortcomings in the machine performance allowing for remedial action to be taken on evidence based data. PMID- 15784458 TI - Red cell storage lesion: the potential impact of storage-induced CD47 decline on immunomodulation and the survival of leucofiltered red cells. AB - Red blood cells undergo major biochemical and biomechanical changes during storage that could effect their post transfusion performance. Biochemical effects include changes in 2,3-diphosphoglycerate (2,3-DPG), ATP, and calcium levels, as well as metabolic modulation and release of Annexin V, a cytosolic component of blood cells, as a global marker of cellular injury and fragmentation. Biomechanical changes include alterations in cellular membrane, shape changes, phospholipid content, phospholipid asymmetry, and antigenic markers. Although the extent of these changes under various storage conditions has been well documented, their clinical effects remain unclear. In the current era of universal leucodepletion, the immunomodulatory effects of some essential markers such as CD47 and phosphatidyl serine become the focus of interest as highlighted in this manuscript. PMID- 15784459 TI - CMV-infected allogeneic endothelial cells initiate responder and bystander donor HLA class I release via the metalloproteinase cleavage pathway. AB - Although cytomegalovirus (CMV) interferes with major histocompatibility expression in infected cells, both host and donor soluble human leukocyte antigen class I (sHLA-I) are often released into the serum of transplant recipients during CMV infection and may contribute to anti-HLA antibody production and graft rejection. We hypothesized that CMV infection of endothelial cells (EC) induces host T cells to release interferon (IFN)-gamma, which in turn drives the metalloproteinase (MPase)-cleavage pathway of sHLA-I generation in "bystander" uninfected ECs. To test this hypothesis, cultures of peripheral blood mononuclear cells (PBMCs) and either uninfected ECs or CMV-infected ECs (EC/CMV) were established and supernatants were tested in enzyme-linked immunosorbent assay for sHLA-I. Responder PBMC became activated and released sHLA-I via the MPase pathway when stimulated with allogeneic EC/CMV; the sHLA-I release was contact dependent and cytokine independent. In transwell cultures, IFN-gamma released by PBMCs in response to EC/CMV stimulated a release of sHLA-I from uninfected allogeneic ECs across the transwell; this release was also MPase dependent. This implies that CMV infection within the transplanted allograft will not only stimulate the release of self HLA from responding PBMCs, but will also stimulate the release of donor sHLA-I from uninfected bystander ECs, both via the class I MPase-pathway. PMID- 15784461 TI - Alleviation of chronic GVHD in mice by oral immuneregulation toward recipient pretransplant splenocytes does not jeopardize the graft versus leukemia effect. AB - Chronic graft versus host disease (cGVHD) is the result of an immune-mediated attack by transplanted donor lymphocytes, entailing inflammatory damage to host target organs. Clinically, the post-bone marrow transplantation (BMT) graft versus leukemia (GVL) effect may be associated with GVHD. Immune hyporesponsiveness induced by oral antigen administration has recently been shown to prevent the development of cGVHD in a murine model. To evaluate whether amelioration of cGVHD in mice by induction of oral immune regulation in donors toward recipient pretransplant lymphocyte antigens is associated with attenuation of the GVL effect donor B10.D2 mice were fed with Balb/c splenocytes, B10.D2 splenocytes, bovine serum albumin (BSA), or regular chow, every other day for 10 days. Subsequently, transplantation of 2 x 10(7) splenocytes from donor B10.D2 mice to recipient Balb/c mice was undertaken, followed by inoculation of 3 x 10(3) BCL-1 leukemia on the day of BMT. Control groups were fed identically without leukemia inoculation. Mice were followed for survival and leukemia progression. Induction of tolerance was assessed by a mixed lymphocyte reaction (MLR). Cutaneous GVHD was assessed macroscopically. To elucidate the mechanism of any observed effect, serum interferon (IFN), interleukin (IL-2), IL-12, IL-4, and IL-10 were determined by enzyme-linked immunosorbent assay and flow cytometry analysis for CD4+, CD8+, and NK1.1+ lymphocyte subpopulations was performed. There was no significant difference in leukemia progression manifested by survival or white blood cell counts of orally immune-regulated mice compared with control animals. Cutaneous cGVHD was significantly ameliorated in Balb/c mice transplanted from tolerized B10.D2 mice. This effect was associated with a significant reduction in the mixed lymphocyte response of effector splenocytes from tolerized B10.D2 mice against Balb/c target splenocytes; significantly decreased serum IFN-gamma and IL-2; increased serum IL-12 levels; increased peripheral NK1.1+ cells; and CD4+/CD8+ lymphocyte ratio. Oral tolerization of BMT donors toward recipient antigens ameliorates cGVHD without hampering the GVL effect. PMID- 15784460 TI - Modulation of monocyte/macrophage function by human CD4+CD25+ regulatory T cells. AB - The suppressive effects of CD4+CD25+ regulatory T cells (Tregs) on T cells have been well documented. Here we investigated whether human CD4+CD25+ Tregs can inhibit the proinflammatory properties of monocytes/macrophages. Monocytes and T cells were isolated from peripheral blood of healthy volunteers by magnetic cell separation and cocultured for 40 h. Monocytes were analyzed directly for cytokine production and phenotypic changes or repurified and used in T-cell stimulation and lipopolysaccharide challenge assays. Coculture with CD4+CD25+ Tregs induced minimal cytokine production in monocytes, whereas coculture with CD4+CD25- T cells resulted in large amounts of proinflammatory (tumor necrosis factor-alpha, interferon-gamma, interleukin-6) and regulatory (interleukin-10) cytokines. Importantly, when these CD4+CD25+ Treg-treated monocytes were repurified after coculture and challenged with lipopolysaccharide, they were severely inhibited in their capacity to produce tumor necrosis factor-alpha and interleukin-6 compared with control-treated monocytes. In addition, monocytes that were precultured with CD4+CD25+ Tregs displayed limited upregulation of human leukocyte antigen class II, CD40 and CD80, and downregulation of CD86 compared with control-treated monocytes. This altered phenotype had functional consequences, as shown by the reduction in T cell-stimulatory capacity of Treg-treated monocytes. Together, these data demonstrate that CD4+CD25+ Tregs can exert direct suppressive effects on monocytes/macrophages, thereby affecting subsequent innate and adaptive immune responses. PMID- 15784462 TI - Difference in target cell recognition of naive and activated human natural killer cells: role of Haymaker (p38.5) in tumoricidal activity. AB - The human Haymaker gene, at 19q13.2 in the Leukocyte Receptor-Receptor-Related Complexes, encodes a 38.5-kDa non-MHC protein found on the plasma membrane of tumor cell lines that are highly susceptible to lysis by naive (unstimulated) natural killer (nNK) cells. We hypothesized that Haymaker might act as a ligand in the tumoricidal activity of nNK cells because this molecule was absent from the surface of malignant cell lines that were resistant to their cytolytic activity. We examined the capacity of this protein to act as a ligand in the tumoricidal activity of freshly isolated nNK cells and IL-2 activated natural killer (NK) cells (aNK cells) by performing blocking studies with recombinant Haymaker (r-Haymaker) and peptide-specific anti-Haymaker antibodies. These competition studies demonstrated that both r-Haymaker and anti-Haymaker antibodies inhibited the tumoricidal function of nNK but not aNK cells. We conclude that Haymaker likely plays an essential role as an activating ligand in nNK-cell-mediated cytotoxicity, whereas the tumoricidal activity of aNK cells is, for the most part, induced by other molecules. PMID- 15784463 TI - Reduced immunosuppression in pediatric liver-intestine transplant recipients with CD8+CD28- T-suppressor cells. AB - Twelve pediatric liver (n = 7), liver-kidney (n = 1), and small bowel (n = 4) transplant recipients, median age 6.5 years (1-21), who received rabbit anti human thymocyte globulin (rATG) and steroid-free tacrolimus/sirolimus, were screened for the presence of CD8+CD28- T suppressor cells. Four control liver transplant recipients, median age 15 years (5-20), in whom conventional immunosuppression without rATG was successfully discontinued for at least 1 year, were also screened as a reference population. Median time to CD8+CD28- T suppressor cells analysis was 16 months (2-24) in rATG subjects and 168 months (16-228) in no-immunosuppression subjects. Nine of 16 patients demonstrated the presence of CD8+CD28- T-suppressor cells in the circulation, whereas seven patients did not. CD8+CD28- T-suppressor cells were present in 4/4 children with no immunosuppression, and absent from three of four subjects with acute cellular rejection, all of whom experienced more than one acute cellular rejection episode. In the reduced immunosuppression group (n = 8), four children demonstrated presence of CD8+CD28- T-suppressor cells in the circulation and four did not. The presence of donor-specific T-suppressor cells in the circulation may characterize transplant recipients, in whom graft function can be maintained with minimal or no immunosuppression. Such assays may also permit safe evaluation of prospective immunosuppression withdrawal strategies. PMID- 15784464 TI - Indels and imperfect duplication have driven the evolution of human Complement Receptor 1 (CR1) and CR1-like from their precursor CR1 alpha: importance of functional sets. AB - This study examines the effects of duplication and insertions-deletions (indels) by comparing human complement receptor 1 (CR1) and human CR1-like (CR1L) with syntenic genes from four other vertebrates (chimpanzee, baboon, rat, and mouse). By phylogenetic analysis, the domains of these genes can be classified into 10 distinct subfamilies (a, b, c, d, e, f, g(-like), h, j, and k), which have been largely conserved throughout vertebrate and invertebrate evolution. In spite of many complex and diverse duplications and indels, the subfamily order of domains (a, j, e, f, b, k, d, g(-like)) has been maintained. The number of domain sets has increased progressively, thereby expanding the functional repertoire. PMID- 15784465 TI - Synergistic inhibition of tumor necrosis factor-related apoptosis-inducing ligand induced apoptosis in human pancreatic beta cells by Bcl-2 and X-linked inhibitor of apoptosis. AB - To better understand the cytokine death-signal transduction pathways in human beta cells, we investigated the inhibitory effects of Bcl-2 (protooncogene bcl-2) and X-linked inhibitor of apoptosis (XIAP) on TRAIL (TNF-related apoptosis inducing ligand)-induced human beta-cell destruction. A panel of Bcl-2 overexpressing transfectants of the human beta-cell lines NES2Y and CM was developed by transfection with a pEFpGKpuro vector containing Bcl-2 or an empty vector as a control. TRAIL-induced cytotoxicity and apoptosis of Bcl-2 overexpressing beta cells were clearly decreased, in comparison with wild-type cells and the empty vector transfectants. XIAP-overexpressing CM, NES2Y, and primary islet cells were generated by exposing cells to recombinant adenovirus expressing XIAP (AdXIAP) or AdLacz as a control. TRAIL-induced cytotoxicity and apoptosis of CM, NES2Y, and primary islet cells infected with AdXIAP were clearly reduced compared with controls. Interestingly, cytotoxicity induced by TRAIL in human beta cells transfected with both Bcl-2 and AdXIAP was much less than that observed in human beta cells transfected with either Bcl-2 or XIAP alone (p < 0.005 in CM and p < 0.03 in NES2Y). Overexpression of both Bcl-2 and XIAP inhibited TRAIL-induced activation of caspases as well as TRAIL-mediated damage of mitochondrial function in cells, suggesting possible regulatory mechanisms. These results indicate that Bcl-2 and XIAP synergistically inhibit TRAIL-mediated death pathways in human beta cells. PMID- 15784466 TI - Interaction between KIR3DL1 and HLA-B*57 supertype alleles influences the progression of HIV-1 infection in a Zambian population. AB - KIR and HLA loci are both highly polymorphic, and some HLA class 1 products bind and trigger cell-surface receptors specified by KIR genes. We examined whether KIR genes act in concert with HLA-B locus to control HIV-1 infection in a sample of Zambian patients. DNA samples from 88 Zambian patients with HIV-1 were examined. Patients were classified as either slow progressors (SP; n = 54) or rapid progressors (RP; n = 34) to AIDS. All were typed for HLA-B and KIR genes. Our results reveal an association between B*57 supertype (B*57s, which includes B*57 and B*58 alleles) and delayed progression to AIDS (p = 0.0007 by pc = 0.015; OR = 5.25). We also observed an increase incidence of Bw4-I80 in patients with slow progression (p = 0.001 by pc = 0.003, OR = 5). This increase was found to be secondary to B*57s. The presence of both KIR3DL1 and B*57S has a significant effect on progression to AIDS (p = 0.0008; OR = 5.61). B*57s genotypes with another HLA-B allele different from those in the trans position, which also had a specificity different to Bw4-I80 (Bw4-T80 or Bw6), was also greater in the SP than in the RP group (p = 0.00003; OR = 10.11). The presence of the inhibitory allele KIR3DL1 in combination with the HLA-B*57s alleles that contain the Bw4-I80 epitope, has a highly protective effect against progression to AIDS in Zambian patients. PMID- 15784467 TI - Single nucleotide polymorphisms in the regulatory regions of the HLA-DRB expressed genes. AB - DRB genes encode proteins that play an important role in the immune response, and their expressional regulation is crucial to the immune reaction. Sequence variation at the regulatory region can directly affect the gene expression level. The aim of the present study was to use Chinese samples to investigate the variation in the regulation region of the human leukocyte antigen (HLA)-DRB expressed genes. Seventy- one single nucleotide polymorphisms (SNPs) were found in the four HLA-DRB-expressed genes. By comparing these data with SNPs in the U.S. National Center for Biotechnology Information dbSNP database, 69 SNPs (97.2%) were found to be novel. In addition, two genetic variations of insertion deletion polymorphisms were discovered within the regulatory region of HLA-DRB1 gene. These polymorphisms can be used as resources of markers for association studies of complex diseases, for assessment of individual predisposition to diseases, and as research markers for population genetics and evolution. PMID- 15784468 TI - Human leukocyte antigens in Japanese patients with biliary atresia: retrospective analysis of patients who underwent living donor liver transplantation. AB - Biliary atresia (BA) is a neonatal obstructive cholangiopathy characterized by a fibrosclerosing obliteration of the extrahepatic bile duct. The aim of this study was to investigate the relationship between human leukocyte antigens (HLA) and susceptibility to BA. We retrospectively analyzed 392 Japanese patients with BA and without extrahepatic anomalies who underwent living donor liver transplantations at our institute. Healthy Japanese volunteers (n = 828) served as normal controls. A significant positive association was observed between BA and HLA-DR2 (39.0% of patients vs. 30.4% of controls, odds ratio = 1.46, p = 0.029). Two-locus analyses disclosed that DR2 was not independently associated with BA, but the increased frequency of HLA-A24 and -B52 reflected the linkage disequilibrium between -A24, -B52, and -DR2. Moreover, the frequency of the haplotype HLA-A24-B52-DR2 was significantly higher in patients with BA than in the general Japanese populations described in the literature (odds ratio = 2.20, p = 0.00124). These results indicate that the gene for BA susceptibility is in close linkage disequilibrium with the HLA-A24-B52-DR2 haplotype observed in the Japanese population. We speculate that a gene at the locus close to HLA plays an important role in the pathogenesis of BA. PMID- 15784469 TI - Human leukocyte antigen class I B and C loci contribute to Type 1 Diabetes (T1D) susceptibility and age at T1D onset. AB - Alleles of human leukocyte antigen (HLA) class II genes are well known to affect susceptibility to type 1 diabetes (T1D), but less is known about the contribution of HLA class I alleles to T1D susceptibility. In this study, molecular genotyping was performed at the HLA-B and HLA-C loci for 283 multiplex Caucasian families, previously typed for HLA-A and the class II loci. Allele frequencies were compared between affected siblings and affected family-based controls. Linkage disequilibrium coefficients were calculated for HLA-B-HLA-C haplotypes and for class I-lass II haplotypes. After adjustment for linkage disequilibrium, the following alleles remain associated with T1D: B*1801, B*3906, B*4403, C*0303, C*0802, and C*1601. B and C allele associations were tested for certain T1D associated DRB1-DQB1 haplotypes, with the following results: B*3801 is protective on DRB1*0401-DQB1*0302 haplotypes, both C*0701 and C*0702 are predisposing on DRB1*0404-DQB1*0302 haplotypes, and B*3906 is predisposing on DRB1*0801-DQB1*0402 haplotypes. As with previous results for HLA-A, HLA-B and HLA-C are associated with age at T1D onset (mean 11.6 +/- 0.3 years). The protective allele B*4403 was associated with older age at onset (15.1 years; p < 0.04), and the predisposing alleles C*0702 and B*3906 were associated with younger age at onset (9.5 years, p < 0.001; and 7.8 years, p < 0.002, respectively). These data support a role for HLA class I alleles in susceptibility to and age at onset of T1D. PMID- 15784470 TI - Eotaxin-3 gene polymorphisms are associated with rheumatoid arthritis in a Korean population. AB - The eotaxin gene family (eotaxin, eotaxin-2, and eotaxin-3) has been implicated in the recruitment of eosinophils, basophiles and Th2 lymphocytes that is a central aspect of allergic diseases. We previously suggested that Eo2+179T>C and Eo2+275C>T of the eotaxin-2, and Eo3+2497T>G of the eotaxin-3 were significantly associated with susceptibility to asthma. To precisely determine whether these single nucleotide polymorphisms (SNPs) are associated with susceptibility to autoimmune disease such as rheumatoid arthritis (RA) in Koreans, we analyzed the genotype and allele frequencies for four SNPs (Eo2+179T>C, Eo2+275C>T, Eo2+304A>C, and Eo2+1272A>G) of the eotaxin-2, and three SNPs (Eo3+77C>T, Eo3+1577G>A, and Eo3+2497T>G) of the eotaxin-3 by single-base extension method. Although the genotype and allele frequencies of the eotaxin-2 SNPs gene between patients with RA and controls were not significantly different, the genotype and allele frequencies of the eotaxin-3SNPs between them were significantly associated. The genotype frequencies of Eo3+1577G>A and Eo3+2497T>G in patients with RA were significantly different from those in the controls (p = 0.0001 and p < 0.0001, respectively). Our results strongly suggest that the polymorphisms of eotaxin-3 might be associated with susceptibility to RA. PMID- 15784471 TI - CTLA4/CT60 polymorphism is not relevant in susceptibility to autoimmune inflammatory intestinal disorders. AB - The aim of this work was to investigate the possible influence of the recently described CT60 A/G dimorphism of the CTLA4 (cytotoxic T-lymphocyte antigen 4) gene in the susceptibility to two different autoimmune inflammatory intestinal disorders, inflammatory bowel disease (IBD) and celiac disease. We analyzed a case-control cohort composed of 528 Spanish patients with IBD (284 with Crohn disease and 244 with ulcerative colitis) and 454 unrelated healthy individuals, and additionally a group of 90 celiac disease families. CT60 genotyping was performed with a TaqMan 5' allelic discrimination assay. After comparing patients with IBD with the control population, we found no significant deviation in the distribution of the alleles or genotypes of CTLA4/CT60 dimorphism. In addition, by means of familial and case-control analysis, no evidence for a statistically significant association was observed between CTLA4/CT60 and celiac disease susceptibility. Therefore, our results suggest that the CTLA4/CT60 polymorphism does not play a major role in inflammatory intestinal disorders. PMID- 15784472 TI - Nomenclature for factors of the HLA System, update October 2004. PMID- 15784473 TI - Nomenclature for factors of the HLA System, update November 2004. PMID- 15784474 TI - Nomenclature for factors of the HLA System, update December 2004. PMID- 15784475 TI - Enteral fish oil in acute pancreatitis. PMID- 15784476 TI - Can oxidative damage be treated nutritionally? AB - BACKGROUND & AIMS: Nutrition and dietary patterns have been shown to have direct impact on health of the population and of selected patient groups. The beneficial effects have been attributed to the reduction of oxidative damage caused by the normal or excessive free radical production. The papers aims at collecting evidence of successful supplementation strategies. METHODS: Review of the literature reporting on antioxidant supplementation trials in the general population and critically ill patients. RESULTS: Antioxidant vitamin and trace element intakes have been shown to be particularly important in the prevention of cancer, cardiovascular diseases, age related ocular diseases and in aging. In animal models, targeted interventions have been associated with reduction of tissue destruction is brain and myocardium ischemia-reperfusion models. In the critically ill antioxidant supplements have resulted in reduction of organ failure and of infectious complications. CONCLUSIONS: Antioxidant micronutrients have beneficial effects in defined models and pathologies, in the general population and in critical illness: ongoing research encourages this supportive therapeutic approach. Further research is required to determined the optimal micronutrient combinations and the doses required according to timing of intervention. PMID- 15784477 TI - Polyamines: metabolism and implications in human diseases. AB - Classically, polyamines is a family of molecules (i.e. putrescine, spermine, spermidine) derived from ornithine according to a decarboxylation/condensative process. More recently, it has been demonstrated that arginine can be metabolised according to the same pathway leading to agmatine formation. Polyamines are essential for the growth, the maintenance and the function of normal cells. The complexity of their metabolism and the fact that polyamines homeostasis is tightly regulated support the idea that polyamines are essential to cell survival. Multiple abnormalities in the control of polyamines metabolism might be implicated in several pathological processes. This paper summarises the knowledge about polyamines metabolism and point out the importance of these molecules as a target for the development of therapeutic tools and nutritional strategy. PMID- 15784478 TI - Effect of enterally administered n-3 polyunsaturated fatty acids in acute pancreatitis--a prospective randomized clinical trial. AB - BACKGROUND: In acute pancreatitis (AP) administration of n-3 polyunsaturated fatty acids (PUFAs) might change the course of the disease through modulation of eicosanoid synthesis. PATIENTS AND METHODS: In a prospective, randomized clinical trial from 28 patients with moderate-severe AP, 14 received n-3 PUFAs (fish oil) enterally (3.3g/day for 5-7 days). Measurement of erythrocyte superoxide dysmutase (SOD) activity, serum total antioxidant status (TAS), vitamin A and E, fatty acids, C-reactive protein, transthyretin concentrations were performed at admission, day 3, 7 and 14. RESULTS: The n-3 to n-6 LCPUFA ratios increased significantly in serum lipids of the patients receiving n-3 PUFA supplementation, whereas remained unchanged in the controls. Supplementation resulted in significant decrease in length of hospitalization (13.07+/-6.70 vs. 19.28+/-7.18 days, P<0.05) and jejunal feeding (10.57+/-6.70 vs. 17.57+/-10.52, P<0.05). Complications developed in 6/14 (42%) of treated and 9/14 (64%) of control patients. The SOD activity was significantly higher at day 3 in the supplemented group (P<0.05), but there were no significant differences between the two groups in other antioxidants and acute phase reactants. CONCLUSION: The use of enteral formula enriched with n-3 PUFAs in the treatment of AP seems to have clinical benefits based upon the shortened time of jejunal feeding and hospital stay. PMID- 15784479 TI - Chefs' knowledge of coeliac disease (compared to the public): a questionnaire survey from the United Kingdom. AB - BACKGROUND: For patients with coeliac disease (CD), compliance with a gluten-free diet, when eating food not prepared at home may be difficult. AIMS: We assessed whether there are differences in the eating habits of coeliac patients when compared to the general public. In addition, we compared chefs' knowledge with the public's knowledge about CD. METHOD: A questionnaire survey about CD was performed on coeliac patients, chefs and the general public from the United Kingdom (UK). RESULTS: Three hundred and nineteen coeliac patients [mean age 53.8 years, 65.8% female], 513 members of the public [49.2 years, 62.6% female] and 322 chefs [37.6 years, 15.2% female] were interviewed. Chefs were less likely to have heard of CD when compared to the public (17.1% (55/322) versus 44.2% (227/513), respectively, P<0.0001). Coeliac patients ate less frequently at a friend's house than the general public (P = 0.003). Coeliac patients ate less frequently from take-away establishments (P<0.0001). However, coeliac patients' ate as frequently in restaurants (P = 0.078). CONCLUSIONS: In the UK, chefs appear to know less about CD than the general public. Patients with CD feel justifiably cautious when eating food not prepared at home. Educating chefs about a gluten-free diet may alleviate the social restrictions on coeliac patients. PMID- 15784481 TI - The introduction of a nutrition clinical nurse specialist results in a reduction in the rate of catheter sepsis. AB - BACKGROUND: Catheter-related sepsis remains the major confounding factor in the long-term delivery of TPN. Previous studies have shown that the introduction of clinical nurse specialists (CNS) can lead to a reduction in sepsis from TPN catheters. This retrospective study aimed to determine the impact of a nutrition CNS on the rate of catheter-related sepsis. Furthermore, the cost of providing such a service was examined to see if it was offset from the savings obtained from reducing venous access infection. METHODS: Prior to the employment of a nutrition CNS, the total number of septic catheter complications were retrospectively collected over a 12-month period. Following appointment of the CNS, all patients requiring TPN were prospectively studied for signs of catheter related sepsis and the data collected over a 4-year period. RESULTS: The overall sepsis rate (cumulative percentage) fell significantly in the 4-year period after the nutrition CNS was appointed, from 52% to 2.3%. The reduction in venous catheter access was accompanied by a decrease in cost related to wastage of TPN and insertion of new Hickman lines. CONCLUSION: The role of the CNS is primarily to improve the quality of care provided, but also to provide cost effectiveness. We have demonstrated that in addition to reducing infection rate of central venous catheters; the cost of employing a nutrition CNS is almost completely covered by the savings resulting from the reduction in wasted central venous catheters, TPN and operating time. PMID- 15784480 TI - A prospective randomised trial of probiotics in critically ill patients. AB - BACKGROUND AND AIMS: Probiotics exert a beneficial effect on the host through modulation of gastrointestinal microflora. The aim of this study was to investigate the effect of the probiotic Lactobacillus plantarum 299v on gut barrier function and the systemic inflammatory response in critically ill patients. SUBJECTS AND METHODS: One hundred and three critically ill patients were randomised to receive an oral preparation containing L. plantarum 299v (ProViva) in addition to conventional therapy (treatment group, n = 52) or conventional therapy alone (control group, n = 51). Serial outcome measures included gastric colonisation, intestinal permeability (lactulose/rhamnose dual sugar probe technique), endotoxin exposure (IgM EndoCAb), C-reactive protein and Interleukin 6 levels. RESULTS: L. plantarum had no identifiable effect on gastric colonisation, intestinal permeability, endotoxin exposure or serum CRP levels. There were no differences between the groups in terms of septic morbidity or mortality. On day 15 serum IL-6 levels were significantly lower in the treatment group compared to controls. CONCLUSIONS: The enteral administration of L. plantarum 299v to critically ill patients was associated with a late attenuation of the systemic inflammatory response. This was not accompanied by any significant changes in the intestinal microflora, intestinal permeability, endotoxin exposure, septic morbidity or mortality. PMID- 15784482 TI - A practical posture for hand grip dynamometry in the clinical setting. AB - BACKGROUND & AIMS: The optimal testing position for hand grip strength, which is a useful functional measure of nutritional status, is open to debate. We therefore examined the systematic difference between different postures in order to establish a methodology that is clinically relevant, easy to perform and reproducible. METHODS: Grip strength was measured in the dominant and non dominant hands with a strain gauge dynamometer in three positions: lying at 30 degrees in bed with elbows supported, seated in an armchair with elbows supported and in a chair with elbows unsupported. The average of three readings made in each position, each 1 min apart, was recorded. RESULTS: 55 normally nourished subjects (26 male) were studied. Mean (95% CI) grip strengths measured in the dominant hand with the subject in bed, sitting in an armchair and sitting in a chair were 45.7 (42.3-49.2), 46.3 (42.9-49.8) and 48.5 (45.4-51.7) kg, respectively for males. Corresponding values for females were 29.4 (27.0-31.8), 29.3 (26.8-31.9) and 31.6 (28.8-34.3) kg. There was no significant difference (Student t-paired test) between measurements made in bed and on an armchair (P = 0.49), but the measurements made in a chair were significantly higher than those made in bed (P = 0.001) and in an armchair (P = 0.004). No statistical difference was present, comparing the three separate measurements in each position (Student t-paired test). CONCLUSIONS: Measurement of grip strength using hand dynamometry is reproducible and consistent. As all patients are not able to sit in a chair with elbows unsupported, in clinical practice it is more practicable to perform hand dynamometry with the elbows supported in a bed or armchair. PMID- 15784483 TI - Can home parenteral nutrition be provided by non-specialised centres? 2300 weeks of experience at a district general hospital in the United Kingdom. AB - BACKGROUND & AIMS: Home Parenteral Nutrition (HPN) is an accepted treatment of intestinal failure but is mostly restricted to a few large specialist centres in the UK. The provision of high-quality HPN is of paramount importance to patients with intestinal failure, but its restriction to large specialist centres limits the number of patients who can receive it. The study aim was to determine if HPN can be effectively administered in a non-specialist centre. METHODS: Adult HPN patients at a single District General Hospital in the United Kingdom were analysed by indications, complications and outcome. RESULTS: 2310 patient weeks of HPN were provided to 23 patients, aged 18-80 years with intestinal failure. Catheter infection rate was 0.315 per patient year, with one patient excluded due to persistent nasal digitation. Patients spent 89.96% of their time at home and 82.6% achieved a Karnowsky Index of 70 (generally self-caring or greater). CONCLUSIONS: HPN can be practised at non-specialist District General Hospital level achieving complication rates comparable to large specialist centres, and this lends weight to the argument for a network model to widen provision beyond large tertiary referral specialist centres in the United Kingdom. PMID- 15784484 TI - Free amino acid and glutathione concentrations in muscle during short-term starvation and refeeding. AB - AIM: The effects of short-term starvation and refeeding on the free amino acids and glutathione in skeletal muscle in healthy man are not known. This is necessary baseline knowledge when studying the effects of nutrition, trauma and sepsis on protein, amino acid and glutathione metabolism. METHODS: Concentrations of free amino acids and glutathione in muscle and plasma from young healthy male volunteers (n = 8) were measured before and after a 3-day fast and then again after 2 days refeeding. Nitrogen balance was determined during the study period. RESULTS: The cumulated nitrogen loss was 36.9+/-5.4 g during the fasting period indicating a condition of protein catabolism. During the fasting period decreases were seen in muscle glutamate by 48 +/- 20% and in glutamine by 38 +/- 12%. These changes were returned back to normal levels during the refeeding period. The changes seen in other muscle amino acids during the study period were reflected by similar changes in plasma amino acids, again with normalisation after the refeeding period. Muscle glutathione concentration and the redox status of glutathione remained unaffected of short-term starvation and refeeding. CONCLUSION: A short-term fasting followed by a refeeding period induced changes in the concentrations of concentrations of glutamate, glutamine, branched chained and basic amino acids in muscle and plasma. Despite this, no changes were seen regarding the glutathione levels in muscle and plasma or its redox status, indicating that the glutathione system is of priority. PMID- 15784485 TI - Plasma vitamin B12, folate and homocysteine levels in gastrectomized men. AB - BACKGROUND & AIMS: Hyperhomocysteinemia is a risk factor for cardiovascular disease, dementia and depression. Prevalence rate of hyperhomocysteinemia in gastrectomized patients is not well elucidated. METHODS: We compared plasma total homocysteine, vitamin B12 and folate of gastrectomized male patients with those of control subjects. RESULTS: Total homocysteine was higher in gastrectomized patients than in control subjects (11.7 +/- 4.4 micromol/l vs. 9.3 +/- 2.4 micromol/l, P = .011), while vitamin B12 was lower in patients than in control subjects (382 +/- 211 pg/ml vs. 500 +/- 179 pg/ml, P = .020). Folate of the two groups was comparable (5.8 +/- 2.5 ng/ml vs. 5.9 +/- 2.2 ng/ml). Of 31 gastrectomized patients six (19.4%) showed low vitamin B12 (<233 pg/ml), four (12.9%) low folate (<3.0 ng/ml) and seven (22.6%) hyperhomocysteinemia (>14 micromol/l), whereas they were found in one (3.2%), none (0%) and one (3.2%) of 31 control subjects, respectively. Patients who had undergone gastrectomy because of cancer showed higher total homocysteine compared to patients who had undergone it because of peptic ulcer. Type of surgery did not correlate with total homocysteine in the present study. CONCLUSION: Hyperhomocysteinemia is not rare in male Japanese gastrectomized patients. PMID- 15784486 TI - Handgrip dynamometry in healthy adults. AB - AIMS: To establish normal reference values for handgrip strength in healthy adult subjects using hand dynamometry. SUBJECTS AND METHODS: Descriptive study of 517 healthy volunteers (267 females and 229 males) aged 17-97 years. Grip strength was measured using two different handgrip dynamometers. Three consecutive measurements were made in both the dominant and the non-dominant hands. The results were analysed after stratification by age, gender, weight, height and dominant/non-dominant hand. RESULTS: The handgrip strength depends on gender (non dominant hand: 22.8 +/-7.2 kg in females and 35.1 +/- 12.4 kg in males (mean +/- standard deviation); "t" test P<0.0001) and is negatively correlated with, age (females r = -0.60, males r = -0.67, P<0.01) and positively with height (females r = -0.48, P<0.01, males r = -0.60, P<0.01). CONCLUSIONS: Normal reference values for handgrip strength should be established and stratified according to age and gender. The variations associated with height do not warrant adjustments in relation to this variable. PMID- 15784487 TI - An assay of microsomal membrane-associated proteasomes demonstrates increased proteolytic activity in skeletal muscle of intensive care unit patients. AB - BACKGROUND & AIMS: Muscle wasting during critical illness is generally believed to be an increase in muscle protein degradation mediated by the proteasome proteolytic pathway. Polyubiquinated proteins are recognised and degraded by the 26S multicatalytic proteasome complex. Animal models for various catabolic conditions have shown increased expression of mRNA:s for several enzymes and subunits in the ubiquitin-proteasome pathway as well as an increase in proteasome activity. The aim of this study was to measure the proteolytic activity of the proteasome in human skeletal muscle. We investigated the proteasome activity in leg muscle biopsies from 7 critically ill patients and from a reference group of 7 age and sex matched patients by a method that could also be suitable for repetitive measurements of intensive care unit patients in future studies. METHODS: Proteasomes were isolated by ultracentrifugation and the fractions containing cytosolic soluble and membrane-bound proteasomes were, respectively, incubated with a fluorogenic peptide substrate to assess the chymotrypsin-like peptidase activity. RESULTS: In the critically ill the proteasome activity in the membrane-bound fraction of proteasomes was 30% higher compared to the reference group (P<0.02), whereas no difference was seen regarding the soluble fraction. CONCLUSION: The results indicate that there is an altered distribution of proteasome activity at the subcellular level in skeletal muscle of critically ill intensive care unit patients. PMID- 15784488 TI - Diet energy density and energy intake in palliative care cancer patients. AB - BACKGROUND & AIMS: Anorexia and cachexia are frequent in advanced cancer. Energy density may be important in the control of energy intake; however, current data are derived from studies in healthy or obese populations and there is no agreed definition and methods of calculation. We studied energy density and energy intake in a group of palliative cancer patients and explored which method of energy density calculation that yielded the highest determination coefficient of energy intake. METHODS: Energy density was calculated from 259 food records using four methods, differing in the types of food and beverages included in the analysis. Association between energy density and energy intake was tested using regression analysis. RESULTS: The determination coefficient for total energy intake was: 0.181, 0.148, 0.222, and 0.211 for methods 1-4, respectively (P for all <0.001). In relation to energy per kg body weight and resting energy expenditure, methods 3 and 1 yielded the highest determination coefficient, R2 = 0.164 and 0.184, respectively. CONCLUSIONS: We found a positive association between diet energy density and energy intake. The method used when calculating energy density had limited impact on this association. When calculating energy density all food and beverages should be included in the analysis. PMID- 15784489 TI - Parenteral nutrition with standard solutions in paediatric intensive care patients. AB - BACKGROUND: Paediatric intensive care patients often require parenteral nutrition (PN). Only very few standard mixtures are available for infants and children. Individual PN solutions need to be compounded manually on the ward, if preparation by the hospital pharmacy is not feasible. Since manual compounding is associated with a greater risk of compounding errors and microbial contamination, the use of standard solutions might be a preferable alternative. METHODS: We evaluated the use of standard solutions on the paediatric intensive care unit of the von Hauner Children's Hospital at the University of Munich over a period of 8 months. PN solutions were either prescribed individually or as standard solutions. We evaluated the frequency of standard solution prescriptions and their modification, compared nutrient intakes with standard vs. individual PN solutions as well as the occurrence of laboratory anomalies. RESULTS: Standard PN solutions were prescribed in 68% of cases, individual PN solutions in 32%. Modifications of standard PN solutions were performed in 54%. The intake of a number of macronutrients and electrolytes was similar with individual and standard PN, but calcium and phosphate intakes were lower with individual total PN. Electrolyte imbalances occurred slightly more often with individual PN than with standard PN (34% vs. 26%, respectively). CONCLUSION: Standard PN solutions were used in the majority of patients on a paediatric intensive care unit. We did not detect indications for inadequacy of standard solutions in the majority of patients reviewed. PMID- 15784490 TI - The mode of delivery of parenteral multivitamins influences nutrient handling in an animal model of total parenteral nutrition. AB - BACKGROUND: Very low birthweight preterm infants receive early total parenteral nutrition (TPN) to optimize protein balance. Adding multivitamins (MVP) to the lipid emulsion (MVP+LIP) rather than to the amino acid+dextrose moiety of TPN (AA+MVP) limits the effects of light exposure on lipid peroxidation and vitamin loss. AIM: Compare the effects of the mode of delivery of MVP on nutrient handling and indices of oxidant stress. METHODS: Three-day old guinea pig pups were assigned to TPN containing MVP+amino acids+dextrose+heparin and electrolytes, with lipids provided separately (AA+MVP). Solutions were light exposed (LE, n = 8) or light protected (LP, n = 9). In a further group (n = 7), MVP was co-administered with the lipid moiety and light exposed (LIP+MVP). Variables measured in urine (creatinine, nitrogen, vitamin C) and in liver (protein, glutathione, isoprostane, vitamins A, E, C) were compared by ANOVA. RESULTS: Urinary nitrogen and vitamin C were higher (P<0.05) during LE, while hepatic levels of vitamin C were higher (P<0.05) with LIP+MVP. These results were not related to total peroxide levels in TPN or to markers of oxidant stress. CONCLUSION. Co-administration of MVP with lipid or light protected amino acids offers comparable beneficial effects on nitrogen and vitamin C metabolism. PMID- 15784491 TI - Pre- and postoperative immunonutrition in patients undergoing liver transplantation: a pilot study of safety and efficacy. AB - BACKGROUND & AIMS: Malnutrition is common in patients with end-stage liver disease and is a risk factor for post-transplant morbidity. The goal of this study was to assess the safety of an immune-enhancing diet in patients undergoing liver transplantation and to investigate its effects on nutritional status. METHODS: Fifteen consecutive patients received oral Impact (0.6l/d) for a median 54 (range 10-168)d pre-transplant and enteral Impact was started early after transplant. Total body protein was measured prior to commencing supplemental Impact, immediately prior to transplant and 10, 15, 30, 90, and 180 days post transplant. The results were compared with those from 17 patients who received standard nutritional intervention. RESULTS: All study patients tolerated Impact pre- and postoperatively and there were no safety concerns. Over the preoperative period total body protein increased significantly (P = 0.017). In 7 patients followed for 6 months post-transplant, a significant (P = 0.026) loss of body protein occurred over the first 15 postoperative days which was regained by 6 months. In the patients who did not receive Impact, body protein did not change preoperatively and the loss after surgery was not regained. Infectious complications occurred in 5/15 (33%) Impact patients and 12/17 (71%) non-Impact patients (P = 0.074). CONCLUSIONS: In patients with end-stage liver disease, our results suggest the possibility that Impact may have a role in improving preoperative nutritional status, hastening recovery after transplant, and reducing postoperative infectious complications. These potential benefits need to be confirmed in a randomised controlled trial. PMID- 15784492 TI - Insufficient fluid and energy intake in hospitalised patients with hip fracture. A prospective randomised study of 80 patients. AB - AIMS: Evaluate nutritional status and fluid and energy intake during the first ten days of hospitalisation in a selection of otherwise healthy patients with a hip fracture. METHODS: A prospective randomised controlled study of 80 patients. Nutritional status was assessed at inclusion. The energy and fluid intake was recorded and calculated daily whilst hospitalised. All patients were given ordinary hospital food and beverage. In the treatment group (n = 40) patients also received intraveneous supplementary nutrition (1000 kcal/day) for three days followed by oral supplementary nutrition (400 kcal/day) for seven days or until discharge. RESULTS: One third of patients were classified as malnourished in both groups. The average daily fluid intake/patient was 1300 ml in the control group compared to 1856 ml in the treatment group (P<0.0001). The average daily energy intake/patient was 916 kcal in the control group compared to 1296 kcal in the treatment group (P = 0.003). The mean difference between actual and needed daily fluid intake was -739 ml in the control group and +27 ml in the treatment group (P<0.0001). Corresponding numbers for energy intake was -783 kcal/day in the control group and -228 kcal/day in the treatment group (P = 0.0003). CONCLUSIONS: Malnutrition is common even in a selection of healthy patients with hip fractures. During hospital stay the fluid and energy intake was considerably lower than that needed in the control group. Supplementary nutritional intake for ten days increased the total fluid and energy intake in the treatment group to near needed levels. PMID- 15784493 TI - Impact of long-term HPN on daily life in adults. AB - BACKGROUND AND AIMS: Home parenteral nutrition (HPN) is a therapy that changes life radically and often means lifelong dependence on parenteral feeding. The aim of this study was to gain insight into problems experienced by adult patients who were dependent on long-term HPN. METHODS: A survey was performed on all patients at the ambulatory care clinics of two university centres. We used two techniques: written questionnaires and interviews. The questionnaires addressed fatigue (CIS), quality of sleep, anxiety, depression (BDI), social impairment (subscale SIP68), and sexual functioning. Data were analysed descriptively. Structured interviews inquired about the negative influence of HPN dependence on daily life. These data were quantified by content analysis. RESULTS: The response rate was 76% (n = 48). Questionnaire results: all the respondents had multiple physical symptoms, which they attributed to the underlying disease. Furthermore, severe fatigue (63%), sleeping disorders, (severe) depression (65%), social impairment (55%), and sexual disorders (33%) were present. Quality of life (QoL) correlated with fatigue, sleeping disorders, anxiety, depression, and social impairment (P<0.02). Interview results: psychosocial problems were the main complaints in daily life due to HPN dependence, e.g. negative changes in moods and feelings (including anxiety), lack of freedom, limitations in social life and being dependent. CONCLUSIONS: Although many somatic symptoms were present, HPN dependent patients reported primarily psychosocial problems in daily life. To improve QoL, HPN teams should assess somatic as well as psychosocial aspects standard. Given the large proportion of patients with depressive disorders, therapies such as anti-depressant medication, psychosocial support and cognitive training, should be offered. PMID- 15784494 TI - Clinical evaluation of mastication: validation of video versus electromyography. AB - BACKGROUNDS AND AIMS: The ability to evaluate masticatory function in people with neurological disabilities is important as this function is often compromised in these groups. However, current standard techniques are often impossible in such groups due to cognitive difficulties. This study is a validation of several variables read from standardised video recordings of mastication as indicators of masticatory function. METHODS: Fifteen healthy, fully dentate male subjects were recorded using EMG and by video simultaneously. An evaluation was undertaken of the video parameters (i) to compare their validity against the electromyographic parameters, (ii) to test intra-rater and inter-rater reliability and (iii) to test the ability to discriminate between four model foods differing in hardness. RESULTS: Masticatory time and the number of masticatory cycles counted on video were found to be valid and reliable indicators. In addition, the number of active chewing cycles performed with an open mouth and identification of the chewing side, were found to have reasonable validity and reliability. The former may allow discrimination between food types. CONCLUSION: As an alternative to the complex evaluation of masticatory function, observation of certain parameters from video recording could be an alternative for use in uncooperative patients. PMID- 15784495 TI - Percutaneous radiologic gastrostomy versus nasogastric tube in critically ill patients. AB - AIMS: To examine the feasibility of percutaneous radiologic gastrostomy in critically ill patients and to assess the rates of complications, esophagitis and gastroesophageal reflux when compared with nasogastric tube. METHOD: Sixty patients admitted to a medical intensive care unit and who were supposed to require gastric tubing for at least 14 days were randomized to have a nasogastric tube or a percutaneous radiologic gastrostomy. Patients with gastrostomy contraindication or gastric tubing for more than 2 days were excluded. RESULTS: No major complication requiring invasive treatment was observed. The nasogastric tube was more prone to failure as defined by the impossibility to place or to replace the allocated tube (P = 0.04) and to tube dysfunction (P<0.001), whereas gastrostomy was associated with increased incidence of minor local complications (P<0.001). Ten days after allocation, the rates of esophagitis (15%) and gastroesophageal reflux (24%) were not significantly different between the two groups. CONCLUSION: In selected critically ill patients, percutaneous radiologic gastrostomy carried a low risk of severe complication but we found no benefit in terms of esophagitis and gastroesphageal reflux between early performed gastrostomy and the nasogastric tube. PMID- 15784496 TI - Impact of nutrition route on fungal infections. PMID- 15784498 TI - Where are the genes that cause endometriosis? PMID- 15784499 TI - The molecular pathophysiology of bacterially induced preterm labor: insights from the murine model. AB - Premature delivery, the most important problem in obstetrics in developed countries, continues to vex clinicians and researchers. Despite decades of investigation, the pathophysiology of premature labor is incompletely understood, and therapies or preventive strategies tailored to each of the many potential causes do not exist. The present review addresses one cause of prematurity, namely, intrauterine bacterial infection. Given the vastness of the literature for even this single etiology, we focus on the mouse as a model organism from which much can be learned about mammalian parturition. The underpinnings of bacterially induced labor are believed to involve a signaling cascade that begins with recognition of offending pathogens by cell-surface receptors (toll-like receptors). This cascade then operates through multiple branching and redundant pathways to bring about the changes within the gestational compartment that produce cervical ripening, labor, and ultimately delivery. The major challenge facing researchers is to understand the levels of complexity in the host response, so that prevention and treatment strategies may be sufficiently focused to minimize unwanted side effects, yet sufficiently broad to be effective. Given the complexity of the problem, this understanding can be aided by efficient model systems, of which one in vivo example is the mouse, an organism that shares with humans many similarities in the biochemical and molecular aspects of inflammation induced preterm labor. We propose that tools with the power to assess simultaneously the myriad elements of the hypothesized signaling cascade (ie, genomic and proteomic technologies) are important components of the solution to the puzzle of parturition. PMID- 15784500 TI - Molecular cloning of ovine endothelial nitric oxide synthase and expression in COS-7 cells. AB - While studies of human and bovine endothelial nitric oxide synthase (eNOS) demonstrate activation by Ca(2+)/calmodulin, recent progress demonstrates that eNOS phosphorylation can alter sensitivity to intracellular free calcium ([Ca(2+)](i)). The sheep, however, is widely used as a model for cardiovascular adaptation to pregnancy and ovine uterine artery endothelial cell (UAEC) eNOS undergoes pregnancy-specific (P) enhancement of activity associated with increased Ca(2+) and protein kinase signaling in response to a number of agonists, including adenosine triphosphate (ATP). The degree of homology between the ovine and human full-length cDNAs was not previously known and yet is necessary to determine the validity in using an ovine model to study human physiology. The objectives of this study were to isolate and validate the clone of ovine eNOS cDNA and investigate ovine eNOS activation when expressed in COS-7 cells. The ovine eNOS cDNA has high homology to published human and bovine sequences and shares identity with the bovine amino acid sequence. When ovine eNOS was transiently expressed in COS-7 cells (COS-7/oeNOS), A23187 increased specific catalytic activity in a dose- and time-dependent manner. A23187 stimulated activation of eNOS was, however, also accompanied by phosphorylation of eNOS S1179 and dephosphorylation of T497, demonstrating that an increase in [Ca(2+)](i) may not be the sole mechanism of activation. The physiologic relevance of this was further underscored by the finding that ATP dose dependently increased peak [Ca(2+)](i) and eNOS activity in COS-7/oeNOS, but also increased eNOS p-S1179 and decreased p-T497. This finding was similar to those in ovine P-UAEC treated with the Ca(2+)-mobilizing agonist ATP, wherein activation of eNOS was again concomitant with a rise p-S1179 as well as a slight decrease in p-T497. In conclusion, we describe the full-length ovine eNOS cDNA sequence and show that both physiologic and nonphysiologic calcium-mobilizing agents, which activate ovine eNOS in COS-7 and P-UAEC, do so in association with changes in eNOS phosphorylation. Given this information we can now begin to dissect the relationship between Ca(2+) elevation and specific phosphorylation events in eNOS activation in the ovine model, and thereby gain insight into the possible basis for pregnancy-related dysfunction. PMID- 15784501 TI - Mechanisms of meconium passage: cholinergic stimulation of electromechanical coordination in the fetal colon. AB - OBJECTIVE: Fetal gastrointestinal function develops in utero, with evidence of enhanced motility near-term. Although colonic passage of meconium in utero may be associated with fetal maturation or stress, little is known of the mechanisms potentiating motility. We assessed the effect of bethanechol, a cholinergic prokinetic agent, on colonic muscle muscular contractile and electromyogram (EMG) activity in the near-term ovine fetus. METHODS: Near-term (130 days, n = 8) singleton ovine fetuses were chronically prepared with vascular catheters and three sets of miniature strain gauges and bipolar EMGs on the serosal surface of the transverse colon, left colic flexure, and distal colon. Following a 60-minute control period, fetuses received intravenous bethanechol (60 microg/kg, Low-Beth; 120 microg/kg, High-Beth) at 60 and 180 minutes. Colonic activity was recorded digitally and analyzed for short-duration (2.05). CONCLUSION: In our study, fetal genotype for specific inherited thrombophilias does not appear to be associated with severe preeclampsia. PMID- 15784507 TI - The impact of interpregnancy interval and previous preterm birth on the subsequent risk of preterm birth. AB - OBJECTIVE: To examine the impact of the interpregnancy interval and a previous preterm birth on the subsequent risk of a preterm birth. METHODS: A retrospective analysis was conducted on a group of 4072 women who had at least two consecutive births, excluding multiple gestation, fetal anomalies, cervical incompetence, and stillbirth. Multivariate logistic regression was used to investigate the association between interpregnancy interval, preterm birth of the first child in the pair (index pregnancy), and the risk of a preterm birth of the second child in the pair (outcome pregnancy). RESULTS: Women with interpregnancy intervals of less than 12 months (odds ratio [OR] 1.3; 95% confidence interval [CI] 1.0-1.7) were at increased risks of preterm birth with the outcome pregnancy. Furthermore, there was an increased risk for a subsequent preterm birth in women who had a preterm birth in the index pregnancy (OR 4.2; 95% CI 3.0-6.0). The risk decreased as the interpregnancy interval increased, with a relatively low risk at 18 to 48 months; subsequently, it increased sharply. In contrast, women who had delivered their previous infants at term carried an increased risk of preterm birth with the outcome pregnancy only if the interval was less than 6 months. CONCLUSION: A difference was found in the impact of the interpregnancy interval on the subsequent risk of preterm birth between women with a prior preterm birth and those who previously delivered an infant at term. PMID- 15784508 TI - Lack of association between endometriosis and N-acetyl transferase 1 (NAT1) and 2 (NAT2) polymorphisms in a Japanese population. AB - OBJECTIVE: We investigated the association between endometriosis and polymorphisms in the N-acetyltransferase 1 (NAT1) and N-acetyltransferase 2 (NAT2) genes in a Japanese population, having previously demonstrated a positive association with NAT2 polymorphisms in a UK population. METHODS: Genotyping for NAT1 alleles *3, *4, *10, and *11, and NAT2 alleles *4, *5A, *5B, *5C, *6A, and *7B was performed using polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) and allele-specific PCR (AS-PCR) analysis in 145 ethnically Japanese, endometriosis patients and 182 controls. The NAT1 and NAT2 allele and genotype frequencies were compared in cases and controls using the Fisher exact test. RESULTS: No significant differences between cases and controls were observed in the frequencies of the NAT1 and NAT2 alleles (P = .13; P = .91) and genotypes (P = .24; P = .79), and the NAT2 acetylation phenotypes (P = .46). Dividing the cases into a subgroup, consisting of women with severe disease only (n = 80), had no effect on the results. CONCLUSION: The distribution of NAT1 and NAT2 allele and genotype frequencies were not significantly different between Japanese cases and controls. Our findings suggest that polymorphisms in NAT1 and NAT2 are unlikely to be associated with an increased risk of endometriosis in the Japanese population. PMID- 15784509 TI - PTEN promoter methylation and protein expression in normal early placentas and hydatidiform moles. AB - OBJECTIVE: To investigate the relationship between PTEN promoter methylation and protein expression, and the possible involvement of the PTEN gene in development of gestational trophoblasts and the pathogenesis of hydatidiform moles. METHODS: DNA was extracted from choria of normal early placentas, partial hydatidiform moles, complete hydatidiform moles, and invasive moles, and overdigested by methylation-sensitive endonuclease HpaII. The PTEN promoter was amplificated by polymerase chain reaction. PTEN protein expression was detected by immunohistochemistry. RESULTS: In partial and complete hydatidiform moles, the PTEN promoter methylation rate was significantly higher than in early placentas (72%, 59.4%, 14.3%; P = .000, .002, respectively), and the PTEN protein expression rate was significantly lower than in early placentas (9.1%, 4.5%, 90.5%; P = .000, .000, respectively). However, partial hydatidiform moles, complete hydatidiform moles, and invasive moles were not significant different in terms of PTEN promoter methylation and protein expression. CONCLUSIONS: These findings suggest that the regulation of PTEN expression may play an important role in the development of the early gestational trophoblast and in the pathogenesis of hydatidiform mole, but not in its malignant transformation. PMID- 15784519 TI - Screening and intervention for alcohol and illicit drug abuse:a survey of internal medicine housestaff. AB - This study attempts to determine how internal medicine housestaff screen and intervene for problematic alcohol and illicit drug use, as well as identify factors correlating with favorable practices. A cross-sectional survey was administered to 93 medical housestaff. Of 64 (69%) respondents, 94% reported routinely screening new patients for alcohol or illicit drug use, while only 52% routinely quantified alcohol consumption and 28% routinely used a screening instrument. Housestaff were unfamiliar with national guidelines and felt unprepared to diagnose substance use disorders, particularly prescription drug abuse. Most routinely counseled patients with alcohol (89%) or illicit-drug problems (91%), although only a third of these patients were referred for formal treatment. More thorough screening practices were associated with greater treatment optimism, while favorable referral practices were associated with greater optimism about 12-step program benefit and difficulty with management. These findings suggest areas to be addressed in residency curricula on substance abuse. PMID- 15784521 TI - Opiate dependence treatment with buprenorphine: one year's experience in a family practice residency setting. AB - Buprenorphine became available for office-based treatment of opiate dependence in January 2003, at which time the Underwood-Memorial Hospital Family Practice Residency Program began offering buprenorphine treatment at its family practice center. This article describes the patient selection process, outcomes, and obstacles to treatment. Patients who had a pharmaceutical benefit were much more likely to remain in treatment than those who had to pay for the medication. The authors are not aware of other residency programs currently providing buprenorphine training, and postulate reasons why family physicians in the United States have not readily adopted the office-based opiate treatment model. PMID- 15784520 TI - Effectiveness of bupropion for smoking cessation during pregnancy. AB - Gestational smoking has been established as a risk to the fetus. However, many pregnant smokers are unable to quit due to the highly addictive nature of nicotine. Nicotine replacement therapy has failed in several studies in pregnant women. In a prospective matched, controlled observational study, of 22 pregnant smokers receiving bupropion, 10 (45%) ceased smoking, as compared to 3 (14%) of 22 controls (P = 0.047). Bupropion appears to be effective for smoking cessation therapy during pregnancy. PMID- 15784522 TI - Addressing smoking cessation in methadone programs. AB - Little is known about how well methadone programs address smoking cessation. We describe the frequency of smoking cessation counseling, and factors affecting its provision in methadone programs. We conducted a cross-sectional survey of methadone patients and their counselors. Of 575 patients, 76% were eligible smokers. Although only 48% of patients reported receiving smoking cessation counseling within the previous six months, 97% of counselors reported providing it (p < 0.0001). Time with one counselor was significantly associated with patient report of receiving smoking cessation counseling (OR 1.19 [95% CI 1.04 1.36]). Although addiction counseling is required in methadone programs, nicotine addiction is addressed less than half the time. Methadone programs should prioritize the provision of effective smoking cessation and facilitate continuity of patient-counselor relationships. PMID- 15784523 TI - Naltrexone augments the effects of nicotine replacement therapy in female smokers. AB - BACKGROUND: There is increased recognition that gender differences may influence outcomes and may modify vulnerability to tobacco addiction, severity of course and response to different treatments. We hypothesized that naltrexone, which has been used to successfully treat opioid and alcohol dependence, when combined with nicotine replacement therapy (NRT) and psychosocial therapy (PT) may enhance smoking cessation rates in women. METHODS: Forty-four adult female smokers meeting DSM-IV criteria for nicotine dependence with expired carbon monoxide content of > or = 15 ppm were randomly assigned in a double blind placebo controlled clinical trial of naltrexone 50 mg + NRT patch + psychosocial therapy (N + NRT + PT)(N = 12) or placebo + NRT patch + psychosocial therapy (P + N + PT)(N=12) for 12 weeks. RESULTS: Twelve weeks of treatment was completed by 54.5%. Smoking cessation among females who completed the 12 weeks for N + NRT + PT was 91.7% (11/12) and for P + NRT + PT was 50% (6/12). CONCLUSION: Naltrexone combined with NRT and psychosocial therapy appears to have a positive cessation effect on women and may be a new treatment option for recidivist female smokers. PMID- 15784525 TI - Hepatitis C in alcohol dependence:drinking versus disulfiram. AB - Hepatitis C virus (HCV) infects alcohol dependent (AD) individuals disproportionately. Disulfiram for timely abstinence in HCV + AD cases remains controversial. Our literature review suggests that (1) active drinking accelerates HCV-related liver damage and that abstinence is associated both (2) with a slower course of HCV+ hepatic deterioration and (3) with enhanced response to antiviral HCV treatment. Further, (4) the risk of disulfiram liver injury appears much lower than that from alcohol, (5) HCV+AD individuals require close monitoring during the first 6 months of disulfiram treatment, and (6) early discontinuation of disulfiram usually reverses harmful effects when these occur. Although systematic data are sparse, continued drinking appears much more liver toxic than does disulfiram in this group. Disulfiram therapy may allow (1) prolonged abstinence leading to successful antiviral therapy for HCV, and (2) time to begin behavioral treatments that facilitate long-term abstinence. Sizeable prospective studies of HCV + AD treatment are badly needed. PMID- 15784524 TI - Smoking cessation and alcohol consumption in individuals in treatment for alcohol use disorders. AB - Most individuals with alcohol use disorders are dependent on both alcohol and nicotine, and combined use of both substances is more damaging to health than use of either alone. Although research indicates that alcoholics can quit smoking, discrepant results have been reported regarding whether smoking cessation is associated with increased risk of alcohol relapse. The purpose of this paper was to examine the relationship between smoking cessation and alcohol consumption using data from Project MATCH. Of the 1,307 participants who smoked at any point during the study, 160 (12%) quit. Quitters consumed less alcohol than those who continued smoking. In addition, quitters demonstrated a significant reduction in alcohol consumption at the time of smoking cessation, which was sustained for six months post-cessation. These findings suggest that individuals in treatment for alcohol use disorders who are motivated to stop smoking can safely be encouraged to do so without jeopardizing their sobriety. PMID- 15784526 TI - Methadone maintenance and male sexual dysfunction. AB - PURPOSE: This study reports the prevalence and types of sexual dysfunction in a sample of men on methadone maintenance for opioid dependence, and describes factors which may contribute to sexual dysfunction. METHODS: 92 opioid-dependent men were recruited from a methadone maintenance clinic and completed two questionnaires, a research interview and laboratory measures. RESULTS: Fourteen percent reported some sexual dysfunction. Erectile dysfunction (r = 0.24, p = 0.020), libido dysfunction (r = 0.30, p = 0.003), and global dysfunction (r = 0.26, p = 0.013) increased with increasing age of the patient. Methadone dose showed a significant direct correlation with increased orgasm dysfunction, both before and after adjusting for duration of treatment (p = 0.012). None of the sexual dysfunction subscales or global dysfunction were associated with plasma testosterone or plasma prolactin levels. CONCLUSIONS: The rate of global sexual dysfunction in methadonetreated men is similar to general population studies and should be evaluated using general population guidelines. Orgasm dysfunction is a special case and may respond to methadone dose reduction. PMID- 15784529 TI - Apoplastic oxidation of L-asparagine is involved in the control of the green algal endophyte Acrochaete operculata Correa & Nielsen by the red seaweed Chondrus crispus Stackhouse. AB - Gametophytes of the marine alga Chondrus crispus are more resistant than tetrasporophytes to infection by the filamentous endophytic alga Acrochaete operculata. It has been shown recently that carrageenan oligosaccharides from the resistant gametophytic generation of C. crispus stimulate the secretion of L asparagine (L-Asn) by the endophyte and that the host generates hydrogen peroxide and 2-oxo-succinamic acid after contact with this amino acid. Here the response of C. crispus to L-Asn and its effect on the pathogen is investigated. Chondrus crispus released hydrogen peroxide, ammonium ions, and a carbonyl compound into the medium when exposed to L-Asn. This response was correlated with an increase in oxygen consumption. Inhibitor studies indicated the involvement of a flavoenzyme in the reaction, which was sensitive to high concentrations of the reaction product, ammonium, and to chlorpromazine, quinacrine, and cyanide, inhibitors of L-amino acid oxidase. Cell wall macerate of C. crispus also responded to L-Asn, while protoplasts were inactive. Uptake of L-Asn into the cell was not necessary for the response, suggesting that the involved L-amino acid oxidase is apoplastic. Acrochaete operculata was more sensitive to hydrogen peroxide than C. crispus and settlement of A. operculata zoospores on C. crispus was reduced by 86% in the presence of L-Asn. This reduced settlement could be prevented with catalase. Chondrus crispus thus features an apoplastic amino acid oxidase, which is involved in the control of its endophytic pathogen. The modulation of the amino acid secretion in A. operculata by carrageenan oligosaccharides is therefore a key issue in the etiology of the association. PMID- 15784532 TI - Cloning and sequencing of a genomic island found in the Brazilian purpuric fever clone of Haemophilus influenzae biogroup aegyptius. AB - A genomic island was identified in the Haemophilus influenzae biogroup aegyptius Brazilian purpuric fever (BPF) strain F3031. This island, which was also found in other BPF isolates, could not be detected in non-BPF biogroup aegyptius strains or in nontypeable or typeable H. influenzae strains, with the exception of a region present in the type b Eagan strain. This 34,378-bp island is inserted, in reference to H. influenzae Rd KW20, within a choline transport gene and contains a mosaic structure of Mu-like prophage genes, several hypothetical genes, and genes potentially encoding an Erwinia carotovora carotovoricin Er-like bacteriocin. The product of the tail fiber ORF in the bacteriocin-like region shows a hybrid structure where the C terminus is similar to an H. influenzae phage HP1 tail protein implicating this open reading frame in altering host specificity for a putative bacteriocin. Significant synteny is seen in the entire genomic island with genomic regions from Salmonella enterica subsp. enterica serovar Typhi CT18, Photorhabdus luminescens subsp. laumondii TT01, Chromobacterium violaceum, and to a lesser extent Haemophilus ducreyi 35000HP. In a previous work, we isolated several BPF-specific DNA fragments through a genome subtraction procedure, and we have found that a majority of these fragments map to this locus. In addition, several subtracted fragments generated from an independent laboratory by using different but related strains also map to this island. These findings underscore the importance of this BPF-specific chromosomal region in explaining some of the genomic differences between highly invasive BPF strains and non-BPF isolates of biogroup aegyptius. PMID- 15784533 TI - Chlamydial infection induces pathobiotype-specific protein tyrosine phosphorylation in epithelial cells. AB - Members of the genus Chlamydia are strict obligate intracellular pathogens that exhibit marked differences in host range and tissue tropism despite sharing a remarkable level of genomic synteny. These pathobiotype differences among chlamydiae are also mirrored in their early interactions with cultured mammalian host cells. Chlamydial attachment and entry is known to trigger protein tyrosine phosphorylation. In this study, we examined the kinetics and pattern of protein tyrosine phosphorylation induced by infection with a comprehensive collection of chlamydial strains exhibiting diversity in host, tissue, and disease tropisms. We report new findings showing that protein tyrosine phosphorylation patterns induced by infection directly correlate with the pathobiotype of the infecting organism. Patterns of protein tyrosine phosphorylation were induced following early infection that unambiguously categorized chlamydial pathobiotypes into four distinct groups: (i) Chlamydia trachomatis trachoma biovars (serovars A to H), (ii) C. trachomatis lymphogranuloma venereum biovars (serovars L1 to L3), (iii) C. muridarum, and (iv) C. pneumoniae and C. caviae. Notably, chlamydia-infected murine and human epithelial cells exhibited the same protein tyrosine phosphorylation patterns; this is indirect evidence suggesting that the phosphorylated protein(s) is of chlamydial origin. If our hypothesis is correct, these heretofore-uncharacterized proteins may represent a novel class of bacterial molecules that influence pathogen-host range or tissue tropism. PMID- 15784531 TI - Pleiotropic enhancement of bacterial pathogenesis resulting from the constitutive activation of the Listeria monocytogenes regulatory factor PrfA. AB - Listeria monocytogenes is a facultative intracellular bacterial pathogen that causes serious disease in immunocompromised individuals, pregnant women, and neonates. Bacterial virulence is mediated by the expression of specific gene products that facilitate entry into host cells and enable bacterial replication; the majority of these gene products are regulated by a transcriptional activator known as PrfA. L. monocytogenes strains containing prfA E77K or prfA G155S mutations exhibit increased expression of virulence genes in broth culture and are hypervirulent in mice. To define the scope of the influences of the prfA E77K and prfA G155S mutations on L. monocytogenes pathogenesis, multiple aspects of bacterial invasion and intracellular growth were examined. Enhanced bacterial invasion of host epithelial cells was dependent on the expression of a number of surface proteins previously associated with invasion, including InlA, InlB, and ActA. In addition to these surface proteins, increased production of the hly encoded secreted hemolysin listeriolysin O (LLO) was also found to significantly enhance bacterial invasion into epithelial cell lines for both prfA mutant strains. Although prfA E77K and prfA G155S strains were similar in their invasive phenotypes, the infection of epithelial cells with prfA E77K strains resulted in host cell plasma membrane damage, whereas prfA G155S strains did not alter plasma membrane integrity. Bacterial infection of human epithelial cells, in which the production of LLO is not required for bacterial entry into the cytosol, indicated that prfA E77K cytotoxic effects were mediated via LLO. Both prfA E77K and prfA G155S strains were more efficient than wild-type bacteria in gaining access to the host cell cytosol and in initiating the polymerization of host cell actin, and both were capable of mediating LLO-independent lysis of host cell vacuoles in cell lines for which L. monocytogenes vacuole disruption normally requires LLO activity. These experiments illuminate the diverse facets of L. monocytogenes pathogenesis that are significantly enhanced by the constitutive activation of PrfA via prfA mutations and underscore the critical role of this protein in promoting L. monocytogenes virulence. PMID- 15784530 TI - Apoptosis, pyroptosis, and necrosis: mechanistic description of dead and dying eukaryotic cells. PMID- 15784534 TI - The Actinobacillus actinomycetemcomitans autotransporter adhesin Aae exhibits specificity for buccal epithelial cells from humans and old world primates. AB - Cells of the gram-negative periodontopathogen Actinobacillus actinomycetemcomitans express a surface-exposed, outer membrane autotransporter protein, designated Aae, which has been implicated in epithelial cell binding. We constructed a mutant strain of A. actinomycetemcomitans that contained a transposon insertion in the Aae structural gene (aae) and tested the mutant to determine its ability to bind to buccal epithelial cells (BECs) isolated from healthy volunteers. Significantly fewer mutant cells than wild-type cells bound to BECs. A broad-host-range plasmid that contained an intact aae gene driven by a heterologous tac promoter restored the ability of the mutant strain to bind to BECs at wild-type levels. This plasmid also conferred upon Escherichia coli the ability to express the Aae protein on its surface and to bind to human BECs. Aae expressing E. coli also bound to BECs isolated from six Old World primates but not to BECs isolated from four New World primates or nine other nonprimate mammals, as well as to human gingival epithelial cells but not to human pharyngeal, palatal, tongue, bronchial, or cervical epithelial cells. Our findings indicate that Aae mediates binding of A. actinomycetemcomitans to BECs from humans and Old World primates and that this process may contribute to the host range specificity and tissue tropism exhibited by this bacterium. PMID- 15784535 TI - The TibA adhesin/invasin from enterotoxigenic Escherichia coli is self recognizing and induces bacterial aggregation and biofilm formation. AB - Escherichia coli strains are responsible for many cases of gastrointestinal disease and represent a serious health problem worldwide. An essential step in the pathogenesis of such strains involves recognition and attachment to host intestinal surfaces. TibA is a potent bacterial adhesin associated with a number of enterotoxigenic E. coli strains and mediates bacterial attachment to a variety of human cells; additionally, it promotes invasion of such cells. This adhesin is a surface-displayed autotransporter protein and belongs to the exclusive group of bacterial glycoproteins; only the glycosylated form confers binding to and invasion of mammalian cells. Here we characterized TibA and showed that it possesses self-association characteristics and can mediate autoaggregation of E. coli cells. We demonstrated that intercellular TibA-TibA interaction is responsible for bacterial autoaggregation. Also, TibA expression significantly enhances biofilm formation by E. coli on abiotic surfaces. PMID- 15784536 TI - Biofilm Formation by Neisseria gonorrhoeae. AB - Studies were performed in continuous-flow chambers to determine whether Neisseria gonorrhoeae could form a biofilm. Under these growth conditions, N. gonorrhoeae formed a biofilm with or without the addition of 10 microM sodium nitrite to the perfusion medium. Microscopic analysis of a 4-day growth of N. gonorrhoeae strain 1291 revealed evidence of a biofilm with organisms embedded in matrix, which was interlaced with water channels. N. gonorrhoeae strains MS11 and FA1090 were found to also form biofilms under the same growth conditions. Cryofield emission scanning electron microscopy and transmission electron microscopy confirmed that organisms were embedded in a continuous matrix with membranous structures spanning the biofilm. These studies also demonstrated that N. gonorrhoeae has the capability to form a matrix in the presence and absence of CMP-N-acetylneuraminic acid (CMP-Neu5Ac). Studies with monoclonal antibody 6B4 and the lectins soy bean agglutinin and Maackia amurensis indicated that the predominate terminal sugars in the biofilm matrix formed a lactosamine when the biofilm was grown in the absence of CMP-Neu5Ac and sialyllactosamine in the presence of CMP-Neu5Ac. N. gonorrhoeae strain 1291 formed a biofilm on primary urethral epithelial cells and cervical cells in culture without loss of viability of the epithelial cell layer. Our studies demonstrated that N. gonorrhoeae can form biofilms in continuous-flow chambers and on living cells. Studies of these biofilms may have implications for understanding asymptomatic gonococcal infection. PMID- 15784537 TI - Interactions of Neisseria gonorrhoeae with adherent polymorphonuclear leukocytes. AB - Neisseria gonorrhoeae causes severe exudative urethritis. The exudates from infected individuals contain large numbers of polymorphonuclear leukocytes (PMN) with ingested gonococci. The fate of N. gonorrhoeae within PMN has been a topic of debate for years. In this study, we examined the interactions of N. gonorrhoeae with PMN adherent to surfaces as a system that better models events during clinical disease. Using chemiluminescence to measure reactive oxygen species (ROS), we found that N. gonorrhoeae stimulated PMN to produce a respiratory burst. Different kinetics were seen when PMN were stimulated with opsonized zymosan particles. In addition, ROS were produced predominantly inside the PMN in response to gonococci. Laser scanning confocal microscopy and transmission electron microscopy showed that N. gonorrhoeae rapidly associated with PMN under these experimental conditions and was internalized. Some gonococci were cleared in the first 30 to 60 min after phagocytosis, but a majority of the population persisted for 6 h after phagocytosis. Quantification of viable organisms showed that a significant portion of the population resisted killing. The viability of this subpopulation remained unchanged for 2 h after phagocytosis. A significant increase of viable gonococci from 1 to 6 h was also observed, suggesting intracellular replication. Four different N. gonorrhoeae strains demonstrated the same capacity to resist PMN-mediated killing, whereas Escherichia coli was rapidly killed by PMN under the same conditions. Taken together, these findings suggest that a subpopulation of N. gonorrhoeae resists killing and replicates within PMN phagosomes in spite of NADPH oxidase activation. PMID- 15784538 TI - Mast cells limit systemic bacterial dissemination but not colitis in response to Citrobacter rodentium. AB - Enteropathogenic Escherichia coli and enterohemorrhagic E. coli cause an inflammatory colitis in human patients characterized by neutrophil infiltration, proinflammatory cytokine expression, and crypt hyperplasia. Citrobacter rodentium causes a similar colitis in mice and serves as a model for enteropathogenic E. coli infection in humans. C. rodentium induces systemic T-cell-dependent antibody production that facilitates clearance of the bacteria and protects the host from reinfection. The role of innate immune cells in infectious colitis, however, is less well understood. In this study, we have determined the role of mast cells in the inflammatory response and disease induced by C. rodentium. Mice deficient in mast cells exhibit more severe colonic histopathology and have a higher mortality rate following infection with C. rodentium than do wild-type animals. Despite unimpaired neutrophil recruitment and lymphocyte activation, mast cell-deficient mice have a disseminated infection evident in crucial organ systems that contributes to sepsis. Importantly, mast cells also have the capacity to directly kill C. rodentium. Together, these results suggest that mast cells protect the host from systemic infection by reducing the bacterial load and preventing dissemination of the bacterium from the colon. PMID- 15784539 TI - Fatal Plasmodium falciparum malaria causes specific patterns of splenic architectural disorganization. AB - The spleen is critical for host defense against pathogens, including Plasmodium falciparum. It has a dual role, not only removing aged or antigenically altered erythrocytes from the blood but also as the major lymphoid organ for blood-borne or systemic infections. The human malaria parasite P. falciparum replicates within erythrocytes during asexual blood stages and causes repeated infections that can be associated with severe disease. In spite of the crucial role of the spleen in the innate and acquired immune response to malaria, there is little information on the pathology of the spleen in human malaria. We performed a histological and quantitative immunohistochemical study of spleen sections from Vietnamese adults dying from severe falciparum malaria and compared the findings with the findings for spleen sections from control patients and patients dying from systemic bacterial sepsis. Here we report that the white pulp in the spleens of patients dying from malaria showed a marked architectural disorganization. We observed a marked dissolution of the marginal zones with relative loss of B cells. Furthermore, we found strong HLA-DR expression on sinusoidal lining cells but downregulation on cordal macrophages. P. falciparum infection results in alterations in splenic leukocytes, many of which are not seen in sepsis. PMID- 15784541 TI - Antibiotic-free plasmid stabilization by operator-repressor titration for vaccine delivery by using live Salmonella enterica Serovar typhimurium. AB - Live, attenuated bacteria are effective vectors for heterologous antigen delivery. However, loss of heterologous gene-bearing plasmids is problematic, and antibiotics and their resistance genes are not desirable for in vivo DNA vaccine delivery due to biosafety and regulatory concerns. To solve this problem, we engineered the first vaccine delivery strain that has no requirement for antibiotics or other selectable marker genes to maintain the recombinant plasmid. This model strain of Salmonella enterica serovar Typhimurium, SLDAPD, uses operator-repressor titration (ORT) technology, which requires only the short, nonexpressed lacO sequence for selection and maintenance. SLDAPD, recovered from the spleens and Peyer's patches of mice following oral inoculation, was shown to maintain a plasmid that, in contrast, was lost from parental strain SL3261. We also demonstrated successful application of this technology to vaccine development, since SLDAPD carrying a plasmid without an antibiotic resistance gene that expressed the Yersinia pestis F1 antigen was as efficacious in protecting vaccinated mice against plague as the parental SL3261 strain carrying an antibiotic-selected version of this plasmid. Protection of mice against plague by immunization with Salmonella expressing F1 has previously required two or more doses; here we demonstrated for the first time protective immunity after a single oral immunization. This technology can easily be used to convert any suitable attenuated strain to an antibiotic-free ORT strain for recombinant protein vaccine delivery in humans. PMID- 15784540 TI - Pyrophosphatase of the roundworm Ascaris suum plays an essential role in the worm's molting and development. AB - Previous studies indicated that inorganic pyrophosphatase of Ascaris suum (AsPPase) plays an important role in larval survival in the host. Here we describe a precise role for AsPPase in larval molting and development and also describe the potential role of recombinant AsPPase (rAsPPase) in protective immunity to A. suum infection. Using reverse transcriptase PCR analysis, we found that disruption of AsPPase gene function by RNA interference resulted in suppression of AsPPase mRNA levels. RNA interference also caused inhibition of molting of third-stage larvae (31%) and suppression of native protein expression, as demonstrated by a 56% reduction in enzyme activity and quantified by immunoblot and immunofluorescence analyses, suggesting that AsPPase has a role in the molting process. The anatomic location of the AsPPase native enzyme in the hypodermis of larvae along with its elevated expression prior to and during the molting process supports such a role. Anti-rAsPPase immunoglobulin G (IgG) also resulted in 57% inhibition of molting of A. suum lung-stage third-stage larvae to fourth-stage larvae in vitro with developmental arrest. Antigenic epitopes of AsPPase overlapped the enzyme active sites. Mice immunized with rAsPPase exhibited high antigen-specific IgG antibody responses and were protected (>70%) against a challenge A. suum migratory-phase infection. Splenic T cells from rAsPPase-immunized mice produced low levels of T helper 1-type cytokines (gamma interferon and interleukin-2) in vitro but exhibited an elevated interleukin-10 response. A significantly high level of IgG1 subclass antibodies was found in immunized mice. Our results establish that AsPPase has a critical role in the molting and development of Ascaris roundworms and suggest the potential of AsPPase for use as a candidate vaccine against ascariasis. PMID- 15784542 TI - Melanization of Cryptococcus neoformans affects lung inflammatory responses during cryptococcal infection. AB - The production of melanin pigments is associated with virulence for many microbes. Melanin is believed to contribute to microbial virulence by protecting microbial cells from oxidative attack during infection. However, there is also evidence from various systems that melanins have immunomodulatory properties, which conceivably could contribute to virulence by altering immune responses. To investigate the effect of melanin on the immune response, we compared the murine pulmonary responses to infection with melanized and nonmelanized Cryptococcus neoformans cells. Infection with melanized cells resulted in a greater fungal burden during the early stages of infection and was associated with higher levels of interleukin-4 and MCP-1 and greater numbers of infiltrating leukocytes. Infection with laccase-positive (melanotic) C. neoformans cells also elicited higher MCP-1 levels and more infiltrating leukocytes than did infection with laccase-negative cells. Melanization interfered with phagocytosis in vivo for encapsulated C. neoformans but not for nonencapsulated cells. The results provide strong evidence that cryptococcal melanization can influence the immune response to infection and suggest that immunomodulation is an additional mechanism by which the pigment contributes to virulence. PMID- 15784544 TI - Avoidance of the host immune system through phase variation in Mycoplasma pulmonis. AB - Phase-variable lipoproteins are commonly found in Mycoplasma species. Mycoplasma pulmonis contains a family of extensively studied phase- and size-variable lipoproteins encoded by the vsa locus. The Vsa surface proteins vary at a high frequency, the in vivo significance of which has yet to be determined. We investigated the role of Vsa phase variation in respect to tissue tropism and avoidance of the immune system in the mouse host. Mycoplasmas were cultured 3, 14, and 21 days postinoculation from the nose, lung, trachea, liver, and spleen of experimentally infected C57BL/6 (wild-type), C57BL/6-RAG-1-/- (RAG-/-), and C57BL/6-inducible nitric oxide synthase (iNOS)-/- (iNOS-/-) mice. In wild-type and iNOS-/- mice, a large number of Vsa variants were seen by 21 days postinoculation. In contrast, little Vsa variation occurred in all tissues of RAG /- mice. Analysis of isolates from 14 days postinoculation revealed less variation of the Vsa proteins in iNOS-/- mice than in the wild type. Western blot analysis of isolates from each strain of mouse demonstrated that Vsa phase variation occurred independently of size variation, indicating no obvious selection pressure for size variants. Additionally, these experiments provided no evidence that mycoplasmas producing particular Vsa proteins adhered only to specific tissues. The data strongly indicate that Vsa phase variation is a mechanism for avoidance of the immune system with no obvious contribution to tissue tropism. PMID- 15784543 TI - The Legionella pneumophila tatB gene facilitates secretion of phospholipase C, growth under iron-limiting conditions, and intracellular infection. AB - Our previous mutational analysis of Legionella pneumophila demonstrated a role for type II protein (Lsp) secretion and iron acquisition in intracellular infection and virulence. In gram-negative bacteria, the twin-arginine translocation (Tat) pathway is involved in secretion of proteins, including components of respiratory complexes, across the inner membrane to the periplasm. To assess the significance of Tat for L. pneumophila, tatB mutants were characterized. The mutants exhibited normal growth in standard media but grew slowly under low-iron conditions. They were also impaired in the Nadi assay, indicating that the function of cytochrome c oxidase is influenced by tatB. Consistent with this observation, a subunit of the cytochrome c reductase was shown to be a Tat substrate. Supernatants of the tatB mutants showed a 30% reduction in phospholipase C activity while maintaining normal levels of other Lsp secreted activities. When tested for infection of U937 macrophages, the tatB mutants showed a 10-fold reduction in growth. Double mutants lacking tatB and Lsp secretion were even more defective, suggesting tatB has an intracellular role that is independent of Lsp. tatB mutants were also impaired 20-fold in Hartmannella vermiformis amoebae cultured in the presence of an iron chelator. All mutant phenotypes were complemented by reintroduction of an intact tatB. Thus, L. pneumophila tatB plays a role in the formation of a respiratory complex, growth under low-iron conditions, the secretion of a phospholipase C activity, and intracellular infection. PMID- 15784546 TI - Carbohydrate-binding specificity of the Escherichia coli cytolethal distending toxin CdtA-II and CdtC-II subunits. AB - Intoxication by cytolethal distending toxin depends on assembly of CdtB, the active A component of this AB toxin, with the cell surface-binding (B) component, composed of the CdtA-CdtC heterodimer, to form the active holotoxin. Here we examine the cell surface binding properties of Escherichia coli-derived CdtA-II (CdtA-II(Ec)) and CdtC-II(Ec) and their capacity to provide a binding platform for CdtB-II(Ec). Using a flow cytometry-based binding assay, we demonstrate that CdtB-II(Ec) binds to the HeLa cell surface in a CdtA-II(Ec)- and CdtC-II(Ec) dependent manner and that CdtA-II(Ec) and CdtC-II(Ec) compete for the same structure on the HeLa cell surface. Preincubation of cells with glycoproteins (thyroglobulin and fetuin), but not simple sugars, blocks surface binding of CdtA II(Ec) and CdtC-II(Ec). Moreover, CdtA-II(Ec) and CdtC-II(Ec) bind immobilized fetuin and thyroglobulin as well as fucose and to a lesser degree N acetylgalactoseamine and N-acetylglucoseamine. Removal of N- but not O-linked carbohydrates from fetuin and thyroglobulin prevents binding of CdtA-II(Ec) and CdtC-II(Ec) to these glycoproteins. In addition, removal of N- but not O-linked surface sugar attachments prevents CDT-II(Ec) intoxication. To characterize the cell surface ligand recognized by CdtA-II(Ec) and CdtC-II(Ec), lectins having various carbohydrate specificities were used to block CDT activity and the cell surface binding of CdtA-II(Ec) and CdtC-II(Ec). Pretreatment of cells with AAA, SNA-I, STA, UEA-I, GNA, and NPA partially or completely blocked CDT activity. AAA, EEA, and UEA-I lectins, all having specificity for fucose, blocked surface binding of CdtA-II(Ec) and CdtC-II(Ec). Together, our data indicate that CdtA II(Ec) and CdtC-II(Ec) bind an N-linked fucose-containing structure on HeLa cells. PMID- 15784545 TI - Impact of the SpeB protease on binding of the complement regulatory proteins factor H and factor H-like protein 1 by Streptococcus pyogenes. AB - Microbial pathogens often exploit human complement regulatory proteins such as factor H (FH) and factor H-like protein 1 (FHL-1) for immune evasion. Fba is an FH and FHL-1 binding protein expressed on the surface of the human pathogenic bacterium Streptococcus pyogenes, a common agent of pharyngeal, skin, and soft tissue infections. Fba has been shown to contribute to phagocytosis resistance, intracellular invasion, and virulence in mice. Here, we look at the role of Fba in recruitment of FH and FHL-1 by five serotype M1 isolates of streptococci. Inactivation of fba greatly inhibited binding of FH and FHL-1 by all isolates, indicating that Fba is a major FH and FHL-1 binding factor of serotype M1 streptococci. For three isolates, FH binding was significantly reduced in stationary-phase cultures and correlated with high levels of protease activity and SpeB (an extracellular cysteine protease) protein in culture supernatants. Analysis of a speB mutant confirmed that SpeB accounts for the loss of Fba from the cell surface, suggesting that the protease may modulate FH and FHL-1 recruitment during infection. Comparisons of fba DNA sequences revealed that the FH and FHL-1 binding site in Fba is conserved among the M1 isolates. Although the ligand binding site is not strictly conserved in Fba from a serotype M49 isolate, the M49 Fba protein was found to bind both FH and FHL-1. Collectively, these data indicate that binding of FH and FHL-1 is a conserved function of Fba while modulation of Fba function by SpeB is variable. PMID- 15784547 TI - Redundant roles for Met docking site tyrosines and the Gab1 pleckstrin homology domain in InlB-mediated entry of Listeria monocytogenes. AB - The bacterial pathogen Listeria monocytogenes causes food-borne illnesses leading to gastroenteritis, meningitis, or abortion. Listeria induces its internalization into some mammalian cells through interaction of the bacterial surface protein InlB with host Met receptor tyrosine kinase. Binding of InlB leads to phosphorylation of Met and the adapter Gab1 and to activation of host phosphoinositide (PI) 3-kinase. The mammalian ligand of Met, hepatocyte growth factor, promotes cell motility and morphogenesis in a manner dependent on phosphorylation of two docking site tyrosines at positions 1349 and 1356 in the receptor's cytoplasmic tail. Here we determined if these tyrosines were essential for Listeria entry. A derivative of the human cell line T47D stably expressing a truncated Met lacking most of its cytoplasmic domain was unable to support InlB mediated signaling or entry. Surprisingly, cells expressing mutant Met containing phenylalanine substitutions in both tyrosines 1349 and 1356 (MetYF) allowed entry and InlB-induced Gab1 phosphorylation. However, in contrast to the situation in cells expressing wild-type Met, Gab1 phosphorylation in MetYF cells required PI 3 kinase activity. The Gab1 pleckstrin homology (PH) domain was constitutively associated with the plasma membrane of cells in a PI 3-kinase-dependent manner. Overexpression of the PH domain blocked entry of Listeria into cells expressing MetYF but not into cells expressing wild-type Met. Taken together, these results indicate that the docking site tyrosines are dispensable for internalization when membrane localization of Gab1 is constitutive. Distinct pathways of recruitment by phosphorylated tyrosines in Met and PH domain ligands in the membrane are redundant for bacterial entry. PMID- 15784548 TI - Differential involvement of toll-like receptors 2 and 4 in the host response to acute respiratory infections with wild-type and mutant Haemophilus influenzae strains. AB - We used a mouse model of acute respiratory infections to investigate the role of Toll-like receptor 2 (TLR2) and TLR4 in the host response to Haemophilus influenzae. Acute aerosol exposures to wild-type strains of H. influenzae showed that TLR4 function was essential for TNF-alpha induction, neutrophil influx, and bacterial clearance. To determine how lipooligosaccharide (LOS) modifications would affect the role of TLR4 in inducing the host response, we used acute infections with an H. influenzae strain expressing a mutation in the htrB gene. This mutant strain expresses an LOS subunit with decreased acylation. In response to H. influenzae htrB infection, tumor necrosis factor alpha (TNF-alpha) secretion remained TLR4 dependent. But the decrease in LOS acylation made the neutrophil influx and the bacterial clearance also dependent on TLR2, as shown by the decreased host response elicited in TLR2 knockout mice compared to C57BL/6 mice. A subsequent analysis of TLR2 and TLR4 gene expression by quantitative PCR indicated that TLR4 function induces TLR2 expression and vice versa. These results indicate that some changes in the LOS subunit of H. influenzae can favor signaling through non-TLR4 receptors, such as TLR2. The results also indicate a close interaction between TLR4 and TLR2 that tightly regulates the expression of both receptors. PMID- 15784549 TI - Rhodococcus equi-specific cytotoxic T lymphocytes in immune horses and development in asymptomatic foals. AB - Rhodococcus equi is an important cause of pneumonia in young horses; however, adult horses are immune due to their ability to mount protective recall responses. In this study, the hypothesis that R. equi-specific cytotoxic T lymphocytes (CTL) are present in the lung of immune horses was tested. Bronchoalveolar lavage (BAL)-derived pulmonary T lymphocytes stimulated with R. equi lysed infected alveolar macrophages and peripheral blood adherent cells (PBAC). As with CTL obtained from the blood, killing of R. equi-infected targets by pulmonary effectors was not restricted by equine lymphocyte alloantigen-A (ELA A; classical major histocompatibility complex class I), suggesting a novel or nonclassical method of antigen presentation. To determine whether or not CTL activity coincided with the age-associated susceptibility to rhodococcal pneumonia, CTL were evaluated in foals. R. equi-stimulated peripheral blood mononuclear cells (PBMC) from 3-week-old foals were unable to lyse either autologous perinatal or mismatched adult PBAC targets. The defect was not with the perinatal targets, as adult CTL effectors efficiently killed infected targets from 3-week-old foals. In contrast, significant CTL activity was present in three of five foals at 6 weeks of age, and significant specific lysis was induced by PBMC from all foals at 8 weeks of age. As with adults, lysis was ELA-A unrestricted. Two previously described monoclonal antibodies, BCD1b3 and CD1F2/1B12.1, were used to examine the expression of CD1, a nonclassical antigen presenting molecule, on CTL targets. These antibodies cross-reacted with both foal and adult PBAC. However, neither antibody bound alveolar macrophages, suggesting that the R. equi-specific, major histocompatibility complex unrestricted lysis is not restricted by a surface molecule identified by these antibodies. PMID- 15784550 TI - Campylobacter jejuni-induced cytokine responses in avian cells. AB - Campylobacter jejuni is a major cause of human inflammatory enteritis. During the course of human disease numerous proinflammatory cytokines are produced. Little is known, however, about the cytokine responses produced during the interaction of this bacterium with the avian host. Campylobacter has been considered a commensal of the avian host. Any differences in innate responses to this pathogen between the human and avian hosts should lead to a greater understanding of the disease process in humans. We have demonstrated expression of proinflammatory cytokines and chemokines in response to Campylobacter infection in avian primary chick kidney cells and the avian macrophage cell line HD11. The data indicate that Campylobacter can stimulate the avian host in a proinflammatory manner. The data strongly suggest that the lack of pathology in vivo is not due to an inability of Campylobacter to stimulate a proinflammatory response from avian cells. PMID- 15784551 TI - Interleukin 10- and Fcgamma receptor-deficient mice resolve Leishmania mexicana lesions. AB - Infection of C57BL/6 (B6) mice with Leishmania mexicana is associated with a minimal immune response and chronic disease. Here we show that B6 interleukin 10 /- (IL-10-/-) mice resolve their lesions and exhibit increased gamma interferon (IFN-gamma), nitric oxide production, and delayed-type hypersensitivity. This enhanced resistance was dependent upon IL-12p40, since treatment of L. mexicana infected IL-10-/- mice with anti-IL-12p40 monoclonal antibody abrogated healing. Antibody-opsonized L. mexicana induced IL-10 production by B6 macrophages in vitro, implicating antibody binding to Fc receptors as a mechanism involved in IL 10 production in this infection. Furthermore, B6 FcRgamma-/- mice resolve L. mexicana lesions, and lymph node cells from these mice produced less IL-10 and more IFN-gamma than cells from infected wild-type mice. These data demonstrate that removal of IL-10 or FcgammaR leads to resolution of L. mexicana disease and support a model in which ligation of FcgammaR by L. mexicana-bound immunoglobulin G promotes IL-10 production, leading to chronic disease. PMID- 15784552 TI - Process development and analysis of liver-stage antigen 1, a preerythrocyte-stage protein-based vaccine for Plasmodium falciparum. AB - Plasmodium falciparum liver-stage antigen 1 (LSA-1) is expressed solely in infected hepatocytes and is thought to have a role in liver schizogony and merozoite release. Specific humoral, cellular, and cytokine immune responses to LSA-1 are well documented, with epitopes identified that correlate with antibody production, proliferative T-cell responses, or cytokine induction. With the goal of developing a vaccine against this preerythrocyte-stage protein, we undertook the good manufacturing practices (GMP) manufacture of a recombinant LSA-1 construct, LSA-NRC, incorporating the N- and C-terminal regions of the protein and two of the centrally placed 17-amino-acid repeats. To improve the protein yield, a method of codon harmonization was employed to reengineer the gene sequence for expression in Escherichia coli. A 300-liter GMP fermentation produced 8 kg of bacterial cell paste, and a three-step column chromatographic method yielded 8 mg of purified antigen per g of paste. The final bulk protein was >98% pure, demonstrated long-term stability, and contained <0.005 endotoxin units per 50 microg of protein. To accomplish the initial stages of evaluation of this protein as a human-use vaccine against malaria, we immunized rabbits and mice with LSA-NRC in Montanide ISA 720. New Zealand White rabbits and A/J (H-2K) mice produced high-titer antibodies that recognized liver-stage parasites in infected cultured human hepatocytes. Gamma interferon-producing cells, which have been associated with LSA-1-mediated protection, were detected in splenocytes harvested from immunized mice. Finally, sera taken from people living in a region where malaria is holoendemic recognized LSA-NRC by Western blotting. PMID- 15784553 TI - Mode of action of invasion-inhibitory antibodies directed against apical membrane antigen 1 of Plasmodium falciparum. AB - Antibodies against apical membrane antigen 1 (AMA-1) of Plasmodium falciparum inhibit merozoite invasion into erythrocytes. Invasion-inhibitory polyclonal AMA 1 antibodies inhibit secondary proteolytic processing and surface redistribution of AMA-1 on merozoites. We present evidence supporting inhibition of processing and redistribution as probable causes of inhibition of invasion by polyclonal antibodies. Polyclonal anti-AMA-1 was much more inhibitory than monoclonal antibody (MAb) 4G2dc1 in an invasion assay. Although both polyclonal and monoclonal immunoglobulin G (IgG) inhibited secondary processing of the 66-kDa form of AMA-1, only polyclonal IgG caused its anomalous processing, inhibited its redistribution, and cross-linked soluble forms of AMA-1 on merozoites. Moreover, Fab fragments of polyclonal IgG that fail to cross-link did not show the enhancement of inhibitory effect over intact IgG, as observed in the case of Fab fragments of MAb 4G2dc1. We propose that although blocking of biologically important sites is a common direct mode of action of anti-AMA-1 antibodies, blocking of AMA-1 secondary processing and redistribution are additional indirect inhibitory mechanisms by which polyclonal IgG inhibits invasion. We also report a processing inhibition assay that uses a C-terminal AMA-1-specific MAb, 28G2dc1, to detect merozoite-bound remnants of processing (approximately 20 kDa from normal processing to 48 and 44 kDa and approximately 10 kDa from anomalous processing to a 52-kDa soluble form of AMA-1). The ratio of intensity of 10-kDa bands to the sum of 10- and 20-kDa bands was positively correlated with inhibition of invasion by polyclonal antibodies. This assay may serve as an important immunochemical correlate for inhibition of invasion. PMID- 15784554 TI - Epitopes recognized by a nonautoreactive murine anti-N-propionyl meningococcal group B polysaccharide monoclonal antibody. AB - The capsular polysaccharide of Neisseria meningitidis group B (MBPS) is a polymer of alpha (2-->8) N-acetyl neuraminic acid. The polysaccharide is chemically identical to an autoantigen, polysialic acid (PSA), and is a poor immunogen, even when conjugated to protein carriers. Immunization of mice with MBPS-protein conjugate vaccines, in which N-acetyl groups have been replaced by propionyl groups (N-Pr MBPS), elicits serum bactericidal antibodies. A subpopulation of these antibodies do not cross-react with human PSA. The reasons for the increased immunogenicity of N-Pr MBPS and the antigenic targets of the bactericidal nonautoreactive antibodies are unknown. In this study, we investigated the antigenic targets of a protective murine monoclonal antibody (MAb) prepared against a N-Pr MBPS-tetanus toxoid conjugate vaccine. Binding of the MAb to N-Pr MBPS (as demonstrated by an enzyme-linked immunosorbent assay) and bactericidal activity were inhibited by de-N-acetylated MBPS and re-N-acetylated MBPS, which indicate that N-propionyl groups are not obligatory determinants for binding. The results of affinity selection from a preparation of N-Pr MBPS and matrix-assisted laser desorption ionization-time of flight mass spectroscopic analysis indicated that the minimal epitope recognized by the MAb is a MBPS disaccharide containing one de-N-acetylated residue. Thus, the bacterial capsular epitope recognized by this bactericidal, nonautoreactive, anti-group-B MAb likely contains de-N-acetyl residues. PMID- 15784555 TI - A deficiency in gamma interferon or interleukin-10 modulates T-Cell-dependent responses to heat shock protein 60 from Histoplasma capsulatum. AB - Immunization of mice with heat shock protein 60 from Histoplasma capsulatum or a polypeptide from the protein designated F3 confers protection. Vbeta8.1/8.2+ T cells are critically important for the protective efficacy of this antigen. The production of interleukin-10 and gamma interferon following vaccination is essential for efficacy. In this study, we sought to determine whether the absence of either cytokine modified the repertoire of antigen-reactive T cells and whether it altered the functional properties of T cells. Mice lacking gamma interferon or interleukin-10 manifested a skewed repertoire compared to that of wild-type mice. The bias was most marked in gamma interferon-deficient mice and modestly altered in interleukin-10-deficient animals. The altered repertoire in gamma interferon-deficient mice could not be explained at the level of antigen presentation or by the absence of this population from mice. The proportion of T cells from interleukin-10-deficient mice manifesting a Th1 phenotype was greatly increased compared to that from wild-type animals. Transfer of splenocytes from gamma interferon- or interleukin-10-deficient mice immunized with heat shock protein 60 failed to confer protection in T-cell receptor alpha/beta-/- mice. The transfer of T-cell clones that did not produce both cytokines failed to prolong survival in T-cell receptor alpha/beta-/- mice, whereas the clones with the same features that were derived from wild-type mice did. These results indicate that the cytokine milieu influences the shape of the T-cell receptor repertoire and support the importance of gamma interferon and interleukin-10 in the efficacy of heat shock protein 60. PMID- 15784556 TI - Regulation of type III secretion hierarchy of translocators and effectors in attaching and effacing bacterial pathogens. AB - Human enteropathogenic Escherichia coli (EPEC), enterohemorrhagic E. coli (EHEC), and the mouse pathogen Citrobacter rodentium (CR) belong to the family of attaching and effacing (A/E) bacterial pathogens. They possess the locus of enterocyte effacement (LEE) pathogenicity island, which encodes a type III secretion system. These pathogens secrete a number of proteins into culture media, including type III effector proteins and translocators that are required for the translocation of effectors into host cells. Preliminary evidence indicated that the LEE-encoded SepL and Rorf6/SepD may form a molecular switch that controls the secretion of translocators and effectors in CR. Here, we show that SepL and SepD indeed perform this function in A/E pathogens such as EHEC and EPEC. Their sepL and sepD mutants do not secrete translocators but exhibit enhanced secretion of effectors. We demonstrate that SepL and SepD interact with each other and that both SepL and SepD are localized to the bacterial membranes. Furthermore, we demonstrate that culture media influence the type III secretion profile of EHEC, EPEC, and CR and that low-calcium concentrations inhibit secretion of translocators but promote the secretion of effectors, similar to effects on type III secretion by mutations in sepL and sepD. However, the secretion profile of the sepD and sepL mutants is not affected by these culture conditions. Collectively, our results suggest that SepL and SepD not only are necessary for efficient translocator secretion in A/E pathogens but also control a switch from translocator to effector secretion by sensing certain environmental signals such as low calcium. PMID- 15784557 TI - Surfactant protein D is present in human tear fluid and the cornea and inhibits epithelial cell invasion by Pseudomonas aeruginosa. AB - We have previously shown that human tear fluid protects corneal epithelial cells against Pseudomonas aeruginosa in vitro and in vivo and that protection does not depend upon tear bacteriostatic activity. We sought to identify the responsible tear component(s). The hypothesis tested was that collectins (collagenous calcium dependent lectins) were involved. Reflex tear fluid was collected from healthy human subjects and examined for collectin content by enzyme-linked immunosorbent assay (ELISA) and Western blot with antibody against surfactant protein D (SP-D), SP-A, or mannose-binding lectin (MBL). SP-D, but not SP-A or MBL, was detected by ELISA of human reflex tear fluid. Western blot analysis of whole tears and of high-performance liquid chromatography tear fractions confirmed the presence of SP-D, most of which eluted in the same fraction as immunoglobulin A. SP-D tear concentrations were calculated at approximately 2 to 5 microg/ml. Depletion of SP D with mannan-conjugated Sepharose or anti-SP-D antibody reduced the protective effect of tears against P. aeruginosa invasion. Recombinant human or mouse SP-D used alone reduced P. aeruginosa invasion of epithelial cells without detectable bacteriostatic activity or bacterial aggregation. Immunofluorescence microscopy revealed SP-D antibody labeling throughout the corneal epithelium of normal, but not gene-targeted SP-D knockout mice. SP-D was also detected in vitro in cultured human and mouse corneal epithelial cells. In conclusion, SP-D is present in human tear fluid and in human and mouse corneal epithelia. SP-D is involved in human tear fluid protection against P. aeruginosa invasion. Whether SP-D plays other roles in the regulation of other innate or adaptive immune responses at the ocular surface, as it does in the airways, remains to be explored. PMID- 15784558 TI - Lipopolysaccharide preparation extracted from Porphyromonas gingivalis lipoprotein-deficient mutant shows a marked decrease in toll-like receptor 2 mediated signaling. AB - We recently demonstrated that a new PG1828-encoded lipoprotein (PG1828LP) was able to be separated from a Porphyromonas gingivalis lipopolysaccharide (LPS) preparation, and we found that it exhibited strong cell activation, similar to that of Escherichia coli LPS, through a Toll-like receptor 2 (TLR2)-dependent pathway. In order to determine the virulence of PG1828LP toward cell activation, we generated a PG1828-deficient mutant of P. gingivalis strain 381 by allelic exchange mutagenesis using an ermF-ermAM antibiotic resistance cassette. A highly purified preparation of LPS from a PG1828-deficient mutant (DeltaPG1828-LPS) showed nearly the same ladder-like patterns in silver-stained gels as a preparation of LPS from a wild-type strain (WT-LPS), as well as Limulus amoebocyte lysate activities that were similar to those of the WT-LPS preparation. However, the ability of the DeltaPG1828-LPS preparation to activate NF-kappaB in TLR2-expressing cells was markedly attenuated. Cytokine production by human gingival fibroblasts was also decreased in response to the DeltaPG1828 LPS preparation in comparison with the WT-LPS preparation, and the activity was comparable to the stimulation of highly purified lipid A of P. gingivalis by TLR4. Further, lethal toxicity was rarely observed following intraperitoneal injection of the PG1828-deficient mutant into mice compared to that with the wild type strain, while the DeltaPG1828-LPS preparation showed no lethal toxicity. Taken together, these results clearly indicate that PG1828LP plays an essential role in inflammatory responses and may be a major virulence factor of P. gingivalis. PMID- 15784559 TI - The innate immune system is activated by stimulation of vaginal epithelial cells with Staphylococcus aureus and toxic shock syndrome toxin 1. AB - Despite knowledge of the effects of toxic shock syndrome (TSS) toxin 1 (TSST-1) on the adaptive immune system, little is known about stimulation of the innate immune system, particularly epithelial cells. This study investigated the interactions of TSS Staphylococcus aureus and TSST-1 with human vaginal epithelial cells (HVECs) and porcine mucosal surfaces. When cocultured with HVECs for 6 h, TSS S. aureus MN8 proliferated, formed aggregates on the HVEC surfaces, and produced exotoxins. Receptor binding studies showed that 35S-TSST-1 bound to 5 x 10(4) receptors per HVEC, with saturation at 15 min. Affymetrix Human GeneChip U133A microarray analysis determined S. aureus MNSM (100 bacteria/HVEC) caused at least twofold up- or down-regulation of 410 HVEC genes by 6 h; these data were also confirmed with S. aureus MN8. TSST-1 (100 microg/ml) caused up- or down-regulation of 2,386 HVEC genes by 6 h. In response to S. aureus, the HVEC genes most up-regulated compared to those in controls were those coding for chemokines or cytokines--MIP-3alpha, 478-fold; GRO-alpha, 26-fold; GRO-beta, 14 fold; and GRO-gamma, 30-fold--suggesting activation of innate immunity. TSST-1 also caused up-regulation of chemokine/cytokine genes. Chemokine/cytokine gene up regulation was confirmed by enzyme-linked immunosorbent assays measuring the corresponding proteins induced by S. aureus and TSST-1. S. aureus MN8, when incubated with porcine vaginal tissue, increased the flux of 35S-TSST-1 across the mucosal surface. This was accompanied by influx of lymphocytes into the upper layers of the tissue. These data suggest innate immune system activation through epithelial cells, reflected in chemokine/cytokine production and influx of lymphocytes, may cause changes in vaginal mucosa permeability, facilitating TSST 1 penetration. PMID- 15784560 TI - Increased expression of interleukin-5 (IL-5), IL-13, and tumor necrosis factor alpha genes in intestinal lymph cells of sheep selected for enhanced resistance to nematodes during infection with Trichostrongylus colubriformis. AB - Cytokine gene expression in cells migrating in afferent and efferent intestinal lymph was monitored for extended time periods in individual sheep experimentally infected with the nematode Trichostrongylus colubriformis. Animals from stable selection lines with increased levels of either genetic resistance (R) or susceptibility (S) to nematode infection were used. Genes for interleukin-5 (IL 5), IL-13, and tumor necrosis factor alpha (TNF-alpha), but not for IL-4, IL-10, or gamma interferon (IFN-gamma), were consistently expressed at higher levels in both afferent and efferent lymph cells of R sheep than in S sheep. However, only minor differences were observed in the surface phenotypes and antigenic and mitogenic responsiveness of cells in intestinal lymph between animals from the two selection lines. The IL-4 and IL-10 genes were expressed at higher levels in afferent lymph cells than in efferent lymph cells throughout the course of the nematode infection in animals of both genotypes, while the proinflammatory TNF alpha gene was relatively highly expressed in both lymph types. These relationships notwithstanding, expression of the IL-10 and TNF-alpha genes declined significantly in afferent lymph cells but not in efferent lymph cells during infection. Collectively, the results showed that R-line sheep developed a strong polarization toward a Th2-type cytokine profile in immune cells migrating in lymph from sites where the immune response to nematodes was initiated, although the IFN-gamma gene was also expressed at moderate levels. Genes or alleles that predispose an animal to develop this type of response appear to have segregated with the R selection line and may contribute to the increased resistance of these animals. PMID- 15784561 TI - Effect of B7-2 and CD40 signals from activated antigen-presenting cells on the ability of zwitterionic polysaccharides to induce T-Cell stimulation. AB - Carbohydrates have been thought to stimulate immune responses independently of T cells; however, zwitterionic polysaccharides (ZPSs) from the capsules of some bacteria elicit potent CD4+-T-cell responses in vivo and in vitro. We demonstrated that HLA-DR on professional antigen-presenting cells (APCs) is required for ZPS-induced T-cell proliferation in vitro (15). Recently, it was shown that ZPSs are processed to low-molecular-weight carbohydrates by a nitric oxide-mediated mechanism in endosomes and locate in the major histocompatibility complex class II pathway (5, 15). The effect of the ZPS-mediated expression of HLA-DR and costimulatory molecules on the APC and T-cell engagement and subsequent T-cell activation has not been elucidated. Herein, we report that ZPS mediated induction of HLA-DR-surface expression and T-cell proliferation are maximally enhanced after incubation of APCs for 8 h with ZPS. Treatment of APCs with bafilomycin A inhibits the up-regulation of ZPS-mediated HLA-DR surface expression and leads to inhibition of T-cell proliferation. Monoclonal antibodies (MAbs) to the costimulatory molecules B7-2 and CD40L specifically block ZPS mediated T-cell activation, while a MAb to B7-1 does not. Surface expression of B7-2 and B7-1 but not of CD40 is maximally enhanced at 8 to 16 h of treatment of APCs with ZPS. The results demonstrate that the cellular immune response to ZPS depends on the translocation of HLA-DR to the cell surface and requires costimulation via B7-2 and CD40 on activated APCs. The implication is that activation of ZPS-specific T cells requires an orchestrated arrangement of both presenting and costimulatory molecules to form an immunological synapse. PMID- 15784562 TI - Differential effects of prior exposure to environmental mycobacteria on vaccination with Mycobacterium bovis BCG or a recombinant BCG strain expressing RD1 antigens. AB - In silico analysis reveals that most protective antigens expressed by the antituberculous vaccine Mycobacterium bovis BCG (BCG) are conserved in M. avium, supporting the hypothesis that exposure to environmental mycobacteria generates cross-reactive immune responses blocking BCG activity. We investigated the impact of sensitization with M. avium, M. scrofulaceum, or M. vaccae on the protective efficacy of a recombinant BCG strain expressing RD1 antigens (BCG::RD1), using a mouse model of experimental tuberculosis (TB). No evidence that the RD1-encoded antigens ESAT-6, CFP-10, and PPE68 were expressed by these environmental strains could be demonstrated by Western blot analysis. Mice sensitized with each of these strains did not prime cellular immune responses cross-reacting with the immunodominant ESAT-6. Importantly, clearance of BCG::RD1 from the lungs and spleens of mice exposed to each of the environmental strains before vaccination was minimal compared to that of BCG. In mice sensitized with M. avium, increased persistence of BCG::RD1 correlated with stronger antimycobacterial gamma interferon responses and enhanced protection against aerosol infection with M. tuberculosis, compared to BCG. In contrast, animals exposed to M. scrofulaceum or M. vaccae prior to vaccination with BCG or BCG::RD1 were better protected against TB than were the unsensitized controls. Our results suggest that the inhibitory effect of environmental mycobacteria on the protective efficacy of BCG depends critically on the extent of cross-recognition of antigens shared with the vaccine. In hosts sensitized with M. avium, potent immunogenicity of ESAT-6 and increased persistence of BCG::RD1 may allow this recombinant vaccine to overcome preexisting antimycobacterial responses. PMID- 15784563 TI - Yersinia pestis V protein epitopes recognized by CD4 T cells. AB - Pneumonic plague, an often-fatal disease for which no vaccine is presently available, results from pulmonary infection by the bacterium Yersinia pestis. The Y. pestis V protein is a promising vaccine candidate, as V protein immunizations confer to mice significant protection against aerosolized Y. pestis. CD4 T cells play central roles during vaccine-primed immune responses, but their functional contributions to Y. pestis vaccines have yet to be evaluated and optimized. Toward that end, we report here the identification of three distinct epitopes within the Y. pestis V protein that activate CD4 T cells in C57BL/6 mice. To our knowledge, these are the first identified CD4 T-cell epitopes in any Y. pestis protein. The epitopes are restricted by the I-A(b) class II major histocompatibility complex molecule and are fully conserved between Y. pestis, Yersinia pseudotuberculosis, and Yersinia enterocolitica. Immunizing mice with a V protein-containing vaccine or with short peptides containing the identified epitopes primes antigen-specific production of interleukin 2 and gamma interferon by CD4 T cells upon their restimulation in vitro. Consistent with prior studies documenting protective roles for CD4 T cells during Y. enterocolitica infection, vaccinating mice with a 16-amino-acid peptide encoding one of the epitopes suffices to protect against an otherwise lethal Y. enterocolitica challenge. The identification of these epitopes will permit quantitative assessments of V specific CD4 T cells, thereby enabling researchers to evaluate and optimize the contribution of these cells to vaccine-primed protection against pneumonic plague. PMID- 15784564 TI - Natural history of gastric mucosal cytokine expression in Helicobacter pylori gastritis in Mongolian gerbils. AB - Data regarding the chronological changes in gastric mucosal cytokines in the different phases of Helicobacter pylori infection are unavailable. We examined Mongolian gerbils for up to 52 weeks after H. pylori (ATCC 43504) inoculation. Levels of mRNAs of mucosal cytokines (interleukin-1beta [IL-1beta], gamma interferon [IFN-gamma], IL-4, IL-6, and IL-10) were assessed using real-time reverse transcription-PCR. Starting 26 weeks after H. pylori inoculation, two clinicohistologic patterns appeared: gastric ulcers in 32% and hyperplastic polyps in 68% of gerbils. High levels of mucosal IL-1beta mRNA were observed early in the infection, reaching maximum at 4 weeks and then rapidly declining. Mucosal IFN-gamma mRNA also reached maximal levels at 4 weeks but remained high thereafter. Both IL-1beta and IFN-gamma mRNA levels were consistently higher in the pyloric mucosa than in the fundic mucosa. In contrast, IL-4, IL-6, and IL-10 mRNA levels peaked at 8 to 26 weeks and levels were similar in the pyloric mucosa and the fundic mucosa. IFN-gamma mRNA levels were significantly higher in gerbils with ulcers than in those with hyperplastic polyps (median IFN gamma/glyceraldehyde-3-phosphate dehydrogenase ratio x 100,000 = 650 versus 338, respectively [antrum], and 172 versus 40, respectively [corpus]) (P < 0.05). We propose that the different outcomes (e.g., ulcers or hyperplastic polyps) might relate to imbalances among cytokines. PMID- 15784565 TI - Elucidation of the monoclonal antibody 5G8-reactive, virulence-associated lipopolysaccharide epitope of Haemophilus influenzae and its role in bacterial resistance to complement-mediated killing. AB - The phase-variable locus lex2 is required for expression of a Haemophilus influenzae lipopolysaccharide (LPS) epitope of previously unknown structure. This epitope, which is reactive with monoclonal antibody (MAb) 5G8, has been associated with virulence of type b strains. When strain RM118 (from the same source as strain Rd), in which the lex2 locus and MAb 5G8 reactivity are absent, was transformed with lex2 DNA, transformants that were reactive with MAb 5G8 were obtained. Surprisingly, the 5G8 reactivity of these transformants was phase variable, although the lex2 locus lacked tetrameric repeats and was constitutively expressed. This phase variation was shown to be the result of phase-variable expression of phosphorylcholine (PCho) such that MAb 5G8 reacted only in the absence of PCho. Structural analysis showed that, compared to RM118, the lex2 transformant had acquired a tetrasaccharide, Gal-alpha1,4-Gal-beta1,4 Glc-beta1,4-Glc-beta1,4, linked to the proximal heptose (HepI). A terminal GalNAc was detected in a minority of glycoforms. LPS derived from a mutant of RM7004, a virulent type b strain which naturally expresses lex2 and has LPS containing the same tetrasaccharide linked to HepI as the sole oligosaccharide extension from the inner core, confirmed that GalNAc is not a part of the MAb 5G8-reactive epitope. Thus, MAb 5G8 specifically binds to the structure Gal-alpha1,4-Gal beta1,4-Glc-beta1,4-Glc-beta attached via a 1,4 linkage to HepI of H. influenzae LPS, and we show that the ability to synthesize this novel tetrasaccharide was associated with enhanced bacterial resistance to complement-mediated killing. PMID- 15784566 TI - Function of neisserial outer membrane phospholipase a in autolysis and assessment of its vaccine potential. AB - Outer membrane phospholipase A (OMPLA) is an outer membrane-localized enzyme, present in many gram-negative bacterial species. It is implicated in the virulence of several pathogens. Here, we investigated the presence, function, and vaccine potential of OMPLA in the human pathogen Neisseria meningitidis. Immunoblot analysis showed the presence of OMPLA in 28 out of 33 meningococcal strains investigated. The OMPLA-negative strains all contained a pldA gene, but these alleles contained premature stop codons. All six Neisseria gonorrhoeae strains tested, but only two out of seven commensal neisserial strains investigated, expressed OMPLA, showing that OMPLA is expressed by, but not limited to, many pathogenic neisserial strains. The function of OMPLA was investigated by assessing the phenotypes of isogenic strains, expressing no OMPLA, expressing wild-type levels of OMPLA, or overexpressing OMPLA. OMPLA exhibited phospholipase activity against endogenous phospholipids. Furthermore, OMPLA was characterized as an autolysin that acted under specific conditions, such as prolonged growth of the bacteria. The vaccine potential of the protein was investigated by immunizing mice with in vitro refolded, recombinant OMPLA. High levels of antibody titers were obtained, but the murine sera were neither bactericidal nor protective. Also, convalescent patients and vaccinee sera did not contain detectable levels of anti-OMPLA antibodies, indicating that OMPLA may not be sufficiently immunogenic to be included in a meningococcal vaccine. PMID- 15784567 TI - Role of YadA, Ail, and Lipopolysaccharide in Serum Resistance of Yersinia enterocolitica Serotype O:3. AB - Complement attack is a host strategy leading to elimination of pathogens. Yersinia enterocolitica expresses several potential complement resistance factors: the outer membrane proteins YadA and Ail as well as lipopolysaccharide (LPS). To study the contribution of these factors to the survival of Y. enterocolitica serotype O:3 in nonimmune human serum, we constructed 23 mutant strains of Y. enterocolitica O:3 expressing different combinations of YadA, Ail, LPS O antigen, and LPS outer core. Survival of bacteria was analyzed in normal serum (with functional classical, lectin, and alternative complement activation pathways) and EGTA-Mg-treated serum (only alternative pathway functional). Kinetic killing tests revealed that the most potent single-serum resistance factor needed for long-term survival was YadA; Ail was also indispensable, but it provided short-term survival and delayed the bacterial killing. On the contrary, the LPS O antigen and outer core, when in combination with YadA, Ail, or both, had a minor and often negative effect on serum resistance. Bacteria in the exponential phase of growth were more resistant to serum killing than stationary phase bacteria. After exposing bacteria to EGTA-Mg-treated serum, O antigen could prevent deposition of covalently bound C3b on bacteria at 3 min of incubation, even as a single factor. At later time points (15 and 30 min) it had to be accompanied by YadA, Ail, and outer core. In normal serum, the bacteria were less resistant to C3b deposition. However, no direct correlation between the C3 deposition pattern and bacterial resistance was observed. PMID- 15784568 TI - Fluid- or surface-phase human salivary scavenger protein gp340 exposes different bacterial recognition properties. AB - Salivary scavenger receptor cysteine-rich protein gp340 aggregates streptococci and other bacteria as part of the host innate defense system at mucosal surfaces. In this article, we have investigated the properties of fluid-phase gp340 and hydroxylapatite surface-adsorbed gp340 in aggregation and adherence, respectively, of viridans group streptococci (e.g., Streptococcus gordonii and Streptococcus mutans), non-viridans group streptococci (e.g., Streptococcus pyogenes and Streptococcus suis), and oral Actinomyces. Fluid-phase gp340 and surface-phase gp340 bioforms were differentially recognized by streptococci, which formed three phenotypic groupings according to their modes of interaction with gp340. Group I streptococci were aggregated by and adhered to gp340, and group II streptococci preferentially adhered to surface-bound gp340, while group III streptococci were preferentially aggregated by gp340. Each species of Streptococcus tested was found to contain strains representative of at least two of these gp340 interaction groupings. The gp340 interaction modes I to III and sugar specificities of gp340 binding strains coincided for several species. Many gp340 interactions were sialidase sensitive, and each of the interaction modes (I to III) for S. gordonii was correlated with a variant of sialic acid specificity. Adherence of S. gordonii DL1 (Challis) to surface-bound gp340 was dependent upon expression of the sialic acid binding adhesin Hsa. However, aggregation of cells by fluid-phase gp340 was independent of Hsa and involved SspA and SspB (antigen I/II family) polypeptides. Conversely, both gp340-mediated aggregation and adherence of S. mutans NG8 involved antigen I/II polypeptide. Deletion of the mga virulence regulator gene in S. pyogenes resulted in increased cell aggregation by gp340. These results suggest that salivary gp340 recognizes different bacterial receptors according to whether gp340 is present in the fluid phase or surface bound. This phase-associated differential recognition by gp340 of streptococcal species of different levels of virulence and diverse origins may mediate alternative host responses to commensal or pathogenic bacterial phenotypes. PMID- 15784569 TI - The thermophilic, homohexameric aminopeptidase of Borrelia burgdorferi is a member of the M29 family of metallopeptidases. AB - Proteases are implicated in several aspects of the physiology of microorganisms, as well as in host-pathogen interactions. Aminopeptidases are also emerging as novel drug targets in infectious agents. In this study, we have characterized an aminopeptidase from the spirochete Borrelia burgdorferi, the causative agent of Lyme disease. The aminopeptidolytic activity was identified in cell extracts from B. burgdorferi by using the substrate leucine-7-amido-4-methylcoumarin. A protein displaying this activity was purified from B. burgdorferi by a two-step chromatographic procedure, yielding a approximately 300-kDa homo-oligomeric enzyme formed by monomers of approximately 50 kDa. Gel enzymography experiments showed that enzymatic activity depends on the oligomeric structure of the protease but does not involve interchain disulfide bonds. The enzyme was identified by peptide mass fingerprinting as the putative aminopeptidase II of B. burgdorferi, encoded by the gene BB0069. It shares significant identity to members of the M29/T family of metallopeptidase, is sensitive to bestatin, has a neutral pH optimum, and displays maximal activity at 60 degrees C. Its activity is 1.75-fold higher at the temperature of the mammalian host than at that of the insect host of the pathogen. The activity of this thermophilic aminopeptidase of B. burgdorferi (TAP(Bb)) depends on Zn2+, and temperatures over 70 degrees C promoted its inactivation through a transition from the hexameric state to the monomeric state. Since B. burgdorferi is deficient in pathways for amino acid synthesis, TAP(Bb) could play a role in supplying required amino acids. Alternatively, the enzyme could be involved in peptide and/or protein processing. PMID- 15784570 TI - Virulence of Pseudomonas aeruginosa in a murine model of gastrointestinal colonization and dissemination in neutropenia. AB - Pseudomonas aeruginosa bacteremia in cancer patients develops from initial gastrointestinal (GI) colonization with translocation into the bloodstream in the setting of chemotherapy-induced neutropenia and GI mucosal damage. We established a reproducible mouse model of P. aeruginosa GI colonization and systemic spread during neutropenia. Mice received 2 mg of streptomycin/ml of drinking water and 1,500 U of penicillin G/ml for 4 days and then ingested 10(7) CFU of P. aeruginosa per ml of drinking water for 5 days. After GI colonization levels were determined, cyclophosphamide (Cy) was then injected intraperitoneally (i.p.) three times every other day or an antineutrophil monoclonal antibody, RB6-8C5, was injected i.p. once. Dissemination was defined by the presence of P. aeruginosa in spleens of moribund or dead mice. In this mouse model, P. aeruginosa colonizes the GI tract and then disseminates systemically once Cy or RB6-8C5 is administered. The duration and intensity of neutropenia, related to Cy dose, was found to be a means to compare the virulence of different P. aeruginosa strains, as exhibited by comparisons of strains lacking or producing the virulence-enhancing ExoU cytotoxin. The lipopolysaccharide outer core polysaccharide and O side chains were critical in establishing GI colonization, and P. aeruginosa mutants lacking the aroA gene (necessary for synthesizing aromatic amino acids) were able to establish GI colonization but unable to disseminate. Both the colonization and dissemination phases of P. aeruginosa pathogenesis can be studied in this model, which should prove useful for evaluating pathogenesis, therapies, and associated means to control P. aeruginosa nosocomial infections. PMID- 15784571 TI - Role of SraP, a Serine-Rich Surface Protein of Staphylococcus aureus, in binding to human platelets. AB - The binding of bacteria to platelets is a postulated central event in the pathogenesis of infective endocarditis. Platelet binding by Streptococcus gordonii is mediated in large part by GspB, a high-molecular-mass cell wall glycoprotein. Although Staphylococcus aureus has a GspB homolog (SraP), little is known about its function. SraP has a calculated molecular mass of 227 kDa and, like GspB, is predicted to contain an atypical N-terminal signal sequence, two serine-rich repeat regions (srr1 and srr2) separated by a nonrepeat region, and a C-terminal cell wall anchoring motif (LPDTG). To assess whether SraP contributes to platelet binding, we compared the binding to human platelets of S. aureus strain ISP479C and of an isogenic variant (strain PS767) in which sraP had been disrupted by allelic replacement. Platelet binding in vitro by PS767 was 47% +/- 17% (mean +/- standard deviation) lower than that of ISP479C (P < 0.001). In addition, a recombinant fragment of SraP containing srr1 and the nonrepeat region was found to bind platelets directly. Binding was saturable, suggesting a receptor-ligand interaction. When tested in a rabbit model of endocarditis, in which each animal was simultaneously infected with ISP479C and PS767 at a ratio of approximately 1:1, the titers of the mutant strain within vegetations were significantly lower than those of the parent strain at 1 and 24 h postinfection. These results indicate that SraP can mediate the direct binding of S. aureus to platelets and that the platelet-binding domain of this glycoprotein is located within its N-terminal region. Moreover, the expression of SraP appears to be a virulence determinant in endovascular infection. PMID- 15784572 TI - Immunoglobulin G isotype responses to erythrocyte surface-expressed variant antigens of Plasmodium falciparum predict protection from malaria in African children. AB - We assessed immunoglobulin G (IgG) isotype responses to variant surface antigens (VSA) expressed on parasite-infected erythrocytes of a panel of heterologous isolates during and after acute episodes in groups of Gabonese children presenting with either mild or severe Plasmodium falciparum malaria. In the acute and convalescent phases IgG3 and IgG1 anti-VSA antibodies, respectively, predominated. In the absence of infection, the levels of both cytophilic isotypes waned, while those of IgG4 increased, particularly in those admitted with severe malaria. Prospective analyses showed significantly longer delays between malaria attacks associated both (i) with increasing IgG1 responses with specificity for VSA of isolates from children with mild malaria and (ii) with increasing IgG4 responses with specificity for VSA of isolates from children with severe malaria. These findings imply that the predictive value of prospectively measured cross reactive VSA-specific IgG antibodies with respect to protection against malaria in African children depends both on their isotype and on their fine specificity. PMID- 15784573 TI - The natural killer complex regulates severe malarial pathogenesis and influences acquired immune responses to Plasmodium berghei ANKA. AB - The natural killer complex (NKC) is a genetic region of highly linked genes encoding several receptors involved in the control of NK cell function. The NKC is highly polymorphic, and allelic variability of various NKC loci has been demonstrated in inbred mice. Making use of BALB.B6-Cmv1r congenic mice, in which the NKC from disease-susceptible C57BL/6 mice has been introduced into the disease-resistant BALB/c background, we show here that during murine malaria infection, the NKC regulates a range of pathophysiological syndromes such as cerebral malaria, pulmonary edema, and severe anemia, which contribute to morbidity and mortality in human malaria. Parasitemia levels were not affected by the NKC genotype, indicating that control of malarial fatalities by the NKC cells does not operate through effects on parasite growth rate. Parasite-specific antibody responses and the proinflammatory gene transcription profile, as well as the TH1/TH2 balance, also appeared to be influenced by NKC genotype, providing evidence that this region, known to control innate immune responses via NK and/or NK T-cell activation, can also significantly regulate acquired immunity to infection. To date, NKC-encoded innate system receptors have been shown mainly to regulate viral infections. Our data provide evidence for critical NKC involvement in the broad immunological responses to a protozoan parasite. PMID- 15784574 TI - DNA immunization with Na+-K+ ATPase (Sseat-6) induces protective immunity to larval Strongyloides stercoralis in mice. AB - Strongyloides stercoralis causes chronic asymptomatic infections which can be maintained in the human host for many decades. Identification and treatment of S. stercoralis-infected individuals is required because immunosuppression can lead to fatal hyperinfection. In this study, human immunoglobulin G (IgG) that had previously been shown to transfer protective immunity to mice was used to identify potential protective antigens. Three antigens or genes from S. stercoralis larvae were identified as tropomyosin (Sstmy-1), Na+-K+ ATPase (Sseat 6), and LEC-5 (Sslec-5). The genes were cloned into plasmids for DNA immunization, and mice were immunized intradermally with the three plasmids individually in combination with a plasmid containing murine granulocyte macrophage colony-stimulating factor. Only Na+-K+ ATPase induced a significant reduction in larval survival after DNA immunization. Immunization with a combination of all three plasmids, including Na+-K+ ATPase, did not induce protective immunity. Serum from mice immunized with DNA encoding Na+-K+ ATPase was transferred to naive mice and resulted in partial protective immunity. Therefore, DNA immunization with Na+-K+ ATPase induces protective immunity in mice, and it is the first identified vaccine candidate against infection with larval S. stercoralis. PMID- 15784575 TI - Intranasal interleukin-12 treatment for protection against respiratory infection with the Francisella tularensis live vaccine strain. AB - Francisella tularensis is a gram-negative intracellular bacterium that can induce lethal respiratory infection in humans and rodents. However, little is known about the role of innate or adaptive immunity in protection from respiratory tularemia. In the present study, the role of interleukin-12 (IL-12) in inducing protective immunity in the lungs against intranasal infection of mice with the live vaccine strain (LVS) of F. tularensis was investigated. It was found that gamma interferon (IFN-gamma) and IL-12 were strictly required for protection, since mice deficient in IFN-gamma, IL-12 p35, or IL-12 p40 all succumbed to LVS doses that were sublethal for wild-type mice. Furthermore, exogenous IL-12 treatment 24 h before intranasal infection with a lethal dose of LVS (10,000 CFU) significantly decreased bacterial loads in the lungs, livers, and spleens of wild type BALB/c and C57BL/6 mice and allowed the animals to survive infection; such protection was not observed in IFN-gamma-deficient mice. The resistance induced by IL-12 to LVS infection was still observed in NK cell-deficient beige mice but not in CD8-/- mice. These results demonstrate that exogenous IL-12 delivered intranasally can prevent respiratory tularemia through a mechanism that is at least partially dependent upon the expression of IFN-gamma and CD8 T cells. PMID- 15784577 TI - Lipopolysaccharide is transferred from high-density to low-density lipoproteins by lipopolysaccharide-binding protein and phospholipid transfer protein. AB - Lipopolysaccharide (LPS), the major outer membrane component of gram-negative bacteria, is a potent endotoxin that triggers cytokine-mediated systemic inflammatory responses in the host. Plasma lipoproteins are capable of LPS sequestration, thereby attenuating the host response to infection, but ensuing dyslipidemia severely compromises this host defense mechanism. We have recently reported that Escherichia coli J5 and Re595 LPS chemotypes that contain relatively short O-antigen polysaccharide side chains are efficiently redistributed from high-density lipoproteins (HDL) to other lipoprotein subclasses in normal human whole blood (ex vivo). In this study, we examined the role of the acute-phase proteins LPS-binding protein (LBP) and phospholipid transfer protein (PLTP) in this process. By the use of isolated HDL containing fluorescent J5 LPS, the redistribution of endotoxin among the major lipoprotein subclasses in a model system was determined by gel permeation chromatography. The kinetics of LPS and lipid particle interactions were determined by using Biacore analysis. LBP and PLTP were found to transfer LPS from HDL predominantly to low density lipoproteins (LDL), in a time- and dose-dependent manner, to induce remodeling of HDL into two subpopulations as a consequence of the LPS transfer and to enhance the steady-state association of LDL with HDL in a dose-dependent fashion. The presence of LPS on HDL further enhanced LBP-dependent interactions of LDL with HDL and increased the stability of the HDL-LDL complexes. We postulate that HDL remodeling induced by LBP- and PLTP-mediated LPS transfer may contribute to the plasma lipoprotein dyslipidemia characteristic of the acute phase response to infection. PMID- 15784576 TI - Mouse paneth cell secretory responses to cell surface glycolipids of virulent and attenuated pathogenic bacteria. AB - Mouse Paneth cells respond to bacteria and bacterial cell surface antigens by discharging secretory granules into the lumen of small intestinal crypts (T. Ayabe et al., Nat. Immunol. 1:113-118, 2000). To investigate mechanisms regulating these responses, purified surface glycolipid molecules with known acyl chain modifications and attenuated properties were tested for the ability to stimulate Paneth cell secretion. The antigens included lipopolysaccharide (LPS) from wild-type and msbB-null Escherichia coli and phoP-null and phoP-constitutive Salmonella enterica serovar Typhimurium strains, as well as LPS, lipid A, and lipoteichoic acid from Pseudomonas aeruginosa and Listeria monocytogenes grown in Mg2+-limited media. Measurements of total secreted protein, secreted lysozyme, and the bactericidal peptide activities of collected secretions showed that the purified antigens elicited similar secretory responses from Paneth cells in mouse crypts ex vivo, regardless of glycolipid acyl chain modification. Despite their impaired Tlr4 pathway, Paneth cells in ex vivo C3H/HeJ mouse crypts released equivalent amounts of bactericidal peptide activity in response to purified bacterial antigens, including lipid A. Thus, mouse Paneth cells respond equivalently to purified bacterial cell envelope glycolipids, regardless of functional Tlr4, the structural properties of glycolipid acyl chains, or their association with virulence in humans. PMID- 15784578 TI - Gene expression in Porphyromonas gingivalis after contact with human epithelial cells. AB - Porphyromonas gingivalis, a gram-negative oral anaerobe, is strongly associated with adult periodontitis. The adherence of the organism to host epithelium signals changes in both cell types as bacteria initiate infection and colonization and epithelial cells rally their defenses. We hypothesized that the expression of a defined set of P. gingivalis genes would be consistently up regulated during infection of HEp-2 human epithelial cells. P. gingivalis genome microarrays were used to compare the gene expression profiles of bacteria that adhered to HEp-2 cells and bacteria that were incubated alone. Genes whose expression was temporally up-regulated included those involved in the oxidative stress response and those encoding heat shock proteins that are essential to maintaining cell viability under adverse conditions. The results suggest that contact with epithelial cells induces in P. gingivalis stress-responsive pathways that promote the survival of the bacterium. PMID- 15784579 TI - Processing and major histocompatibility complex class II presentation of Legionella pneumophila antigens by infected macrophages. AB - To better understand interactions between the intracellular pathogen Legionella pneumophila and macrophages (Mphis), host and bacterial determinants important for presentation of antigens on major histocompatibility complex class II molecules (MHC-II) were investigated. It was determined that immune CD4 T-cell responses to murine bone marrow-derived Mphis (BMphis) infected with wild-type L. pneumophila were higher than the responses to avirulent dotA mutant bacteria. Although this enhanced response by immune T cells required modulation of vacuole transport mediated by the Dot/Icm system, it did not require intracellular replication of L. pneumophila. Intracellular cytokine staining identified a population of immune CD4 T cells that produced gamma interferon upon incubation with BMphis infected with wild-type L. pneumophila that did not respond to Mphi infection with dotA mutant bacteria. Endocytic processing was required for presentation of L. pneumophila antigens on MHC-II as determined by a defect in CD4 T-cell responses when the pH of BMphi endosomes was neutralized with chloroquine. Investigation of MHC-II presentation of antigens by BMphis infected with L. pneumophila icmR, icmW, and icmS mutants indicated that these mutants have an intermediate presentation phenotype relative to those of wild-type and dotA mutant bacteria. In addition, it was found that antigens from dot and icm mutants are presented earlier than antigens from wild-type L. pneumophila. Although immune CD4 T-cell responses to proteins secreted by the L. pneumophila Lsp system were not detected, it was found that the Lsp system is important for priming L. pneumophila-specific T cells in vivo. These data indicate that optimal antigen processing and MHC-II presentation to immune CD4 T cells involves synthesis of L. pneumophila proteins in an endoplasmic reticulum-derived compartment followed by transport to lysosomes. PMID- 15784581 TI - Identification and functional characterization of complement regulator-acquiring surface protein 1 of the Lyme disease spirochetes Borrelia afzelii and Borrelia garinii. AB - Complement regulator-acquiring surface protein 1 (CRASP-1) is the dominant factor H-like protein 1 (FHL-1)- and factor-H-binding protein of Borrelia burgdorferi and is suggested to contribute to persistence of the pathogen. The prototype CRASP-1 of B. burgdorferi sensu stricto (CRASP-1Bb) has been formerly characterized. As shown recently, serum-resistant Borrelia afzelii strains express a unique FHL-1 and factor H-binding protein, designated CRASP-1Ba. Here, we describe for the first time the isolation and functional characterization of the gene encoding the full-length CRASP-1Ba of 28 kDa, which, upon processing, is predicted to be 26.4 kDa. CPASP-1Ba of B. afzelii spirochetes is associated with a genetic locus encoding the orthologous gbb54 gene family that maps to the linear plasmid of approximately 54 kb. Ligand affinity blotting techniques demonstrate that both native and recombinant CRASP-1Ba molecules strongly bind to FHL-1 and much more weakly to factor H. The FHL-1 and factor-H-binding site in CRASP-1Ba is shown to be localized to a 12-amino-acid residue domain at the C terminus of the protein. For comparison, the corresponding cspA-like gene(s) of a serum-sensitive Borrelia garinii strain has also been cloned and characterized. Most notably, two CRASP-1-related B. garinii proteins were identified; however, both molecules bind only weakly to FHL-1 and not at all to factor H. The present identification of the binding site of CRASP-1Ba represents an important step forward in our understanding of the pathogenesis of Lyme disease and may be helpful to design therapeutic regimens to interfere with complement evasion strategies of human pathogenic Borrelia strains. PMID- 15784580 TI - Identification and functional characterization of chicken toll-like receptor 5 reveals a fundamental role in the biology of infection with Salmonella enterica serovar typhimurium. AB - Toll-like receptors (TLRs) are a major component of the pattern recognition receptor repertoire that detect invading microorganisms and direct the vertebrate immune system to eliminate infection. In chickens, the differential biology of Salmonella serovars (systemic versus gut-restricted localization) correlates with the presence or absence of flagella, a known TLR5 agonist. Chicken TLR5 (chTLR5) exhibits conserved sequence and structural similarity with mammalian TLR5 and is expressed in tissues and cell populations of immunological and stromal origin. Exposure of chTLR5+ cells to flagellin induced elevated levels of chicken interleukin-1beta (chIL-1beta) but little upregulation of chIL-6 mRNA. Consistent with the flagellin-TLR5 hypothesis, an aflagellar Salmonella enterica serovar Typhimurium fliM mutant exhibited an enhanced ability to establish systemic infection. During the early stages of infection, the fliM mutant induced less IL 1beta mRNA and polymorphonuclear cell infiltration of the gut. Collectively, the data represent the identification and functional characterization of a nonmammalian TLR5 and indicate a role in restricting the entry of flagellated Salmonella into systemic sites of the chicken. PMID- 15784582 TI - Clinical and microbiological responses of volunteers to combined intranasal and oral inoculation with a Streptococcus gordonii carrier strain intended for future use as a group A streptococcus vaccine. AB - Streptococcus gordonii shows promise as a live mucosal vaccine vector for immunization against respiratory pathogens. In preparation for clinical trials to evaluate S. gordonii engineered to express group A streptococcal M protein antigens, we characterized the responses of 150 healthy volunteers to combined nasal and oral inoculation with approximately 1.5 x 10(9) CFU of SP204(1-1), an S. gordonii strain not bearing vaccine antigens. SP204(1-1) was selected for resistance to streptomycin and 5-fluoro-2-deoxyuridine to distinguish it from indigenous flora. In two antibiotic treatment studies, we performed serial culturing of nose, mouth, and saliva samples from 120 subjects treated with azithromycin beginning 5 days after inoculation to determine whether SP204(1-1) could be rapidly eliminated should safety concerns arise. A natural history study was performed to assess the time until spontaneous eradication in the remaining 30 subjects, who did not receive the antibiotic and who were monitored with repeated culturing for 14 weeks after inoculation. SP204(1-1) was generally well tolerated. Symptoms reported most often within 5 days of inoculation were nasal congestion (36%), headache (30%), and sore throat (19%). The strain was detected by culturing in 98% of subjects. A single dose of azithromycin eliminated colonization in 95% of subjects; all subjects receiving a 5-day course of an antibiotic showed clearance by day 11. Without the antibiotic, 82% of subjects showed spontaneous eradication of the implanted strain within 7 days, and all showed clearance by 35 days. The results of these clinical trials provide encouragement that the use of S. gordonii as a live mucosal vaccine vector is a feasible strategy. PMID- 15784584 TI - Generation of attenuated Mycobacterium bovis strains by signature-tagged mutagenesis for discovery of novel vaccine candidates. AB - Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex, has a particularly wide host range and causes tuberculosis in most mammals, including humans. A signature tag mutagenesis approach, which employed illegitimate recombination and infection of guinea pigs, was applied to M. bovis to discover genes important for virulence and to find potential vaccine candidates. Fifteen attenuated mutants were identified, four of which produced no lesions when inoculated separately into guinea pigs. One of these four mutants had nine deleted genes including mmpL4 and sigK and, in guinea pigs with aerosol challenge, provided protection against tuberculosis at least equal to that of M. bovis BCG. Seven mutants had mutations near the esxA (esat-6) locus, and immunoblot analysis of these confirmed the essential role of other genes at this locus in the secretion of EsxA (ESAT-6) and EsxB (CFP10). Mutations in the eight other attenuated mutants were widely spread through the chromosome and included pks1, which is naturally inactivated in clinical strains of M. tuberculosis. Many genes identified were different from those found by signature tag mutagenesis of M. tuberculosis by use of a mouse infection model and illustrate how the use of different approaches enables identification of a wider range of attenuating mutants. PMID- 15784583 TI - Involvement of the Escherichia coli O157:H7(pO157) ecf operon and lipid A myristoyl transferase activity in bacterial survival in the bovine gastrointestinal tract and bacterial persistence in farm water troughs. AB - Escherichia coli O157:H7 is an important food-borne pathogen that causes hemorrhagic colitis and the hemolytic-uremic syndrome in humans. Recently, we reported that the pO157 ecf (E. coli attaching and effacing gene-positive conserved fragments) operon is thermoregulated by an intrinsically curved DNA and contains the genes for bacterial surface-associated proteins, including a second copy of lipid A myristoyl transferase, whose chromosomal copy is the lpxM gene product. E. coli O157:H7 survives and persists well in diverse environments from the human and bovine gastrointestinal tracts (GIT) to nutrient-dilute farm water troughs. Transcriptional regulation of the ecf operon by intrinsic DNA curvature and the genetic redundancy of lpxM that is associated with lipid A modification led us to hypothesize that the pO157 ecf operon and lpxM are associated with bacterial survival and persistence in various in vivo and ex vivo environments by optimizing bacterial membrane structure and/or integrity. To test this hypothesis, three isogenic ecf operon and/or lpxM deletion mutants of E. coli O157:H7 ATCC 43894 were constructed and analyzed in vitro and in vivo. The results showed that a double mutant carrying deletions in the ecf and lpxM genes had an altered lipid A structure and membrane fatty acid composition, did not survive passage through the bovine GIT, did not persist well in farm water troughs, had increased susceptibility to a broad spectrum of antibiotics and detergents, and had impaired motility. Electron microscopic analyses showed gross changes in bacterial membrane structure. PMID- 15784585 TI - Haemophilus ducreyi Outer membrane determinants, including DsrA, define two clonal populations. AB - The Haemophilus ducreyi outer membrane component DsrA (for ducreyi serum resistance A) is necessary for complete resistance to normal human serum (NHS). When DsrA expression in 19 temporally and geographically diverse clinical isolates of H. ducreyi was examined by Western blotting, 5 of the strains expressed a different immunotype of the DsrA protein (DsrA(II)) than the well characterized prototypical strain 35000HP (DsrA(I)). The predicted DsrA proteins expressed by the DsrA(II) strains were 100% identical to each other but only 48% identical to that of strain 35000HP. In addition to the DsrA(II) protein, class II strains also expressed variant forms of other outer membrane proteins (OMPs) including NcaA (necessary for collagen adhesion A), DltA (ducreyi lectin A), Hlp (H. ducreyi lipoprotein), major OMP, and/or OmpA2 (for OMP A2) and synthesized a distinct, faster-migrating lipooligosaccharide. Based on these data, strains expressing DsrA(I) were termed class I, and those expressing DsrA(II) were termed class II. Expression of dsrA(II) from strain CIP 542 ATCC in the class I dsrA(I) mutant FX517 (35000HP background), which does not express a DsrA protein, rendered this strain resistant to 50% NHS. This demonstrates that DsrA(II) protein is also critical to serum resistance. Taken together, these results indicate that there are two clonal populations of H. ducreyi. The implications of two classes of H. ducreyi strains differing in important antigenic outer membrane components are discussed. PMID- 15784586 TI - The UspA2 protein of Moraxella catarrhalis is directly involved in the expression of serum resistance. AB - Many strains of Moraxella catarrhalis are resistant to the bactericidal activity of normal human serum. Previous studies have shown that mutations involving the insertion of an antibiotic resistance cartridge into the M. catarrhalis uspA2 gene resulted in the conversion of a serum-resistant strain to a serum-sensitive phenotype. In the present study, the deletion of the entire uspA2 gene from the serum-resistant M. catarrhalis strain O35E resulted in a serum-sensitive phenotype and did not affect either the rate of growth or the lipooligosaccharide expression profile of this mutant. Inactivation of the classical complement pathway in normal human serum with Mg2+ and EGTA resulted in the survival of this uspA2 mutant. In contrast, blocking of the alternative complement pathway did not protect this uspA2 mutant from complement-mediated killing. To determine whether the UspA2 protein is directly involved in serum resistance, transformation and allelic exchange were used to replace the uspA2 gene in the serum-resistant strain O35E with the uspA2 gene from the serum-sensitive M. catarrhalis strain MC317. The resultant O35E transformant exhibited a serum-sensitive phenotype. Similarly, when the uspA2 gene from the serum-resistant strain O35E was used to replace the uspA2 gene in the serum-sensitive strain MC317, the MC317 transformant acquired serum resistance. The use of hybrid O35E-MC317 uspA2 genes showed that the N-terminal half of the O35E protein contained a 102-amino-acid region that was involved in the expression of serum resistance. In addition, when the uspA2 genes from strains O35E and MC317 were cloned and expressed in Haemophilus influenzae DB117, only the O35E UspA2 protein caused a significant increase in the serum resistance of the H. influenzae recombinant strain. These results prove that the UspA2 protein is directly involved in the expression of serum resistance by certain M. catarrhalis strains. PMID- 15784587 TI - Staphylococcus aureus deficient in lipidation of prelipoproteins is attenuated in growth and immune activation. AB - A lipoprotein diacylglyceryl transferase (lgt) deletion mutant of Staphylococcus aureus SA113 was constructed. The lipoprotein and prelipoprotein expression, the growth behavior, and the ability of the mutant to elicit an immune response in various host cells were studied. In the wild type, the majority of [14C]palmitate labeled lipoproteins were located in the membrane fraction, although some lipoproteins were also present on the cell surface and in the culture supernatant. The lgt mutant completely lacked palmitate-labeled lipoproteins and released high amounts of some unmodified prelipoproteins, e.g., the oligopeptide binding protein OppA, the peptidyl-prolyl cis-trans isomerase PrsA, and the staphylococcal iron transporter SitC, into the culture supernatant. The growth of the lgt mutant was hardly affected in rich medium but was retarded under nutrient limitation. The lgt mutant and its crude lysate induced much fewer proinflammatory cytokines and chemokines in human monocytic (MonoMac6), epithelial (pulmonary A549), and endothelial (human umbilical vein endothelial) cells than the wild type. However, in whole blood samples, the culture supernatant of the lgt mutant was equal or even superior to the wild-type supernatant in tumor necrosis factor alpha induction. Lipoprotein fractionation experiments provided evidence that a small proportion of the mature lipoproteins are released by the S. aureus wild type despite the lipid anchor and are trapped in part by the cell wall, thereby exposing the immune-activating lipid structure on the cell surface. Bacterial lipoproteins appear to be essential for a complete immune stimulation by gram-positive bacteria. PMID- 15784588 TI - Genetic diversity and carriage dynamics of Neisseria lactamica in infants. AB - Neisseria lactamica, a harmless human commensal found predominantly in the upper respiratory tracts of infants, is closely related to Neisseria meningitidis, a pathogen of global significance. Colonization with N. lactamica may be responsible for the increase in immunity to meningococcal disease that occurs during childhood, when rates of meningococcal carriage are low. This observation has led to the suggestion that N. lactamica whole cells or components are potential constituents of novel meningococcal vaccines. However, the dynamics of carriage and population diversity of N. lactamica in children are poorly understood, presenting difficulties for the choice of representative isolates for use in vaccine development. This problem was addressed by the multilocus sequence typing of N. lactamica isolates from two longitudinal studies of bacterial carriage in infants. The studies comprised 100 and 216 subjects, with N. lactamica carriage monitored from age 4 weeks until age 96 weeks and from age 2 weeks until age 24 weeks, respectively. The maximum observed carriage rate was 44% at 56 weeks of age, with isolates obtained on multiple visits for the majority (54 of 75, 72%) of carriers. The N. lactamica isolates were genetically diverse, with 69 distinct genotypes recovered from the 75 infants. Carriage was generally long-lived, with an average rate of loss of under 1% per week during the 28 weeks following acquisition. Only 11 of the 75 infants carried more than one genotypically unique isolate during the course of the study. Some participants shared identical isolates with siblings, but none shared identical isolates with their parents. These findings have implications for the design of vaccines based on this organism. PMID- 15784589 TI - Role of predicted transmembrane domains for type III translocation, pore formation, and signaling by the Yersinia pseudotuberculosis YopB protein. AB - YopB is a 401-amino-acid protein that is secreted by a plasmid-encoded type III secretion system in pathogenic Yersinia species. YopB is required for Yersinia spp. to translocate across the host plasma membrane a set of secreted effector proteins that function to counteract immune signaling responses and to induce apoptosis. YopB contains two predicted transmembrane helices (residues 166 to 188 and 228 to 250) that are thought to insert into the host plasma membrane during translocation. YopB is also required for pore formation and host-cell-signaling responses to the type III machinery, and these functions of YopB may also require membrane insertion. To elucidate the importance of membrane insertion for YopB function, YopB proteins containing helix-disrupting double consecutive proline substitutions in the center of each transmembrane domain were constructed. Yersinia pseudotuberculosis strains expressing the mutant YopB proteins were used to infect macrophages or epithelial cells. Effector translocation, pore formation, and host-cell-signaling responses were studied. Introduction of helix disrupting substitutions into the second transmembrane domain of YopB resulted in a nonfunctional protein that was not secreted by the type III machinery. Introduction of helix-disrupting substitutions into the first transmembrane domain of YopB resulted in a protein that was fully functional for secretion and for interaction with YopD, another component of the translocation machinery. However, the YopB protein with helix-disrupting substitutions in the first transmembrane domain was partially defective for translocation, pore formation, and signaling, suggesting that all three functions of YopB involve insertion into host membrane. PMID- 15784590 TI - Functional analysis of Plasmodium falciparum apical membrane antigen 1 utilizing interspecies domains. AB - Plasmodium falciparum apical membrane antigen 1 (AMA1) is a leading malaria vaccine candidate whose function has not been unequivocally defined. Partial complementation of function can be achieved by exchanging the AMA1 of P. falciparum (PfAMA1) with that of P. chabaudi (PcAMA1). In this study, parasites expressing chimeric AMA1 proteins were created to identify domains of PfAMA1 critical in erythrocyte invasion and which are important immune targets. We report that specific chimeric AMA1 proteins containing domains I to III from PfAMA1 and PcAMA1 were able to complement PfAMA1 function in erythrocyte invasion. We demonstrate that domain III does not contain dominant epitope targets of antibodies raised against Escherichia coli expressed and refolded PfAMA1 ectodomain. Furthermore, we generated a parasite line in which the N terminal pro region of PfAMA1 does not undergo proteolytic cleavage and show that its removal is necessary for PfAMA1 function. PMID- 15784591 TI - Human leukocyte antigen-DQ8 transgenic mice: a model to examine the toxicity of aerosolized staphylococcal enterotoxin B. AB - Staphylococcal enterotoxins (SEs) belong to a large group of bacterial exotoxins that cause severe immunopathologies, especially when delivered as an aerosol. SEs elicit the release of lethal amounts of cytokines by binding to major histocompatibility complex (MHC) class II and cross-linking susceptible T-cell receptors. Efforts to develop effective therapeutic strategies to protect against SEs delivered as an aerosol have been hampered by the lack of small animal models that consistently emulate human responses to these toxins. Here, we report that human leukocyte antigen-DQ8 (HLA-DQ8) transgenic (Tg) mice, but not littermate controls, succumbed to lethal shock induced by SEB aerosols without potentiation. Substantial amounts of perivascular edema and inflammatory infiltrates were noted in the lungs of Tg mice, similar to the pathology observed in nonhuman primates exposed by aerosol to SEB. Furthermore, the observed pathologies and lethal shock correlated with an upsurge in proinflammatory cytokine mRNA gene expression in the lungs and spleens, as well as with marked increases in the levels of proinflammatory circulating cytokines in the Tg mice. Unlike the case for littermate controls, telemetric evaluation showed significant hypothermia in Tg mice exposed to lethal doses of SEB. Taken together, these results show that this murine model will allow for the examination of therapeutics and vaccines developed specifically against SEB aerosol exposure and possibly other bacterial superantigens in the context of human MHC class II receptors. PMID- 15784592 TI - Enterococcus faecalis tropism for the kidneys in the urinary tract of C57BL/6J mice. AB - Enterococcus faecalis is a gram-positive bacterium that can cause a variety of nosocomial infections of which urinary tract infections are the most common. These infections can be exceptionally difficult to treat because of drug resistance of many E. faecalis isolates. Despite their troublesome nature, little is known about the host or bacterial factors necessary for E. faecalis to cause disease in the urinary tract. Using a mouse model of urinary tract infection, we have shown that E. faecalis is capable of persisting in the kidneys of mice for at least 2 weeks. In contrast, bacterial titers from the bladders of the same mice were inconsistent and tended to be much lower than those recovered from the kidney. This preference for the kidney over the bladder is also observed in other clinical E. faecalis strains. Histologic examination of bladder and kidney tissues demonstrated that E. faecalis induced an inflammatory response in the kidney but not in the bladder. This inflammatory response was TLR2 independent and did not induce inflammatory markers typically associated with uropathogenic Escherichia coli. Using a competition assay, we demonstrated that a pyelonephritis clinical isolate had a growth advantage over a laboratory strain of E. faecalis in the kidneys but not in the bladders of mice. Taken together, these results demonstrate that E. faecalis has tropism for the kidneys in the urinary tracts of mice and that this system can be used to study factors involved in the pathogenesis of urinary tract infections. PMID- 15784593 TI - Q fever pneumonia: virulence of Coxiella burnetii pathovars in a murine model of aerosol infection. AB - Q fever is a worldwide zoonosis caused by Coxiella burnetii, a strictly intracellular bacterium that is a potential bioweapon. Humans usually acquires Q fever after inhalation of dust infected by subclinical animals. We used an aerosol exposure apparatus to challenge immunocompetent (BALB/c) and severe combined immunodeficient (SCID) mice with two different strains (strain Nine Mile and strain Q 212) of C. burnetii at two different inocula. Pathological lesions and dissemination of the bacteria were related to the size of the inoculum. SCID mice showed major pulmonary lesions, whereas similarly infected BALB/c mice were more able to eliminate the bacteria. Pathological differences were found between the strains, with Nine Mile showing more severe histological lesions and quantified spread of bacteria. Our animal model could provide a new tool for the study of acute Q fever pneumonia, the development of Q fever in immunodeficient hosts, and the differentiation of pathogenicity among C. burnetii isolates. PMID- 15784594 TI - Heterologous immunity in the absence of variant-specific antibodies after exposure to subpatent infection with blood-stage malaria. AB - We examined immunity induced by subpatent blood-stage malaria (undetectable by microscopy) using the rodent malaria parasite, Plasmodium chabaudi chabaudi, postulating that limited infection may allow expansion of antigen-specific T cells that are normally deleted by apoptosis. After three infections drug cured at 48 h, mice were protected against high-dose challenge with homologous or heterologous parasites (different strain or variant). Immunity differed from that generated by three untreated, patent infections. Subpatently infected mice lacked immunoglobulin G (IgG) to variant surface antigens, despite producing similar titers of total malaria-specific IgG to those produced by patently infected mice, including antibodies specific for merozoite surface antigens conserved between heterologous strains. Antigen-specific proliferation of splenocytes harvested prechallenge was significantly higher in subpatently infected mice than in patently infected or naive mice. In subpatently infected mice, lymphoproliferation was similar in response to homologous and heterologous parasites, suggesting that antigenic targets of cell-mediated immunity were conserved. A Th1 cytokine response was evident during challenge. Apoptosis of CD4+ and CD8+ splenic lymphocytes occurred during patent but not subpatent infection, suggesting a reason for the relative prominence of cell-mediated immunity after subpatent infection. In conclusion, subpatent infection with blood stage malaria parasites induced protective immunity, which differed from that induced by patent infection and targeted conserved antigens. These findings suggest that alternative vaccine strategies based on delivery of multiple parasite antigens at low dose may induce effective immunity targeting conserved determinants. PMID- 15784595 TI - Purification and partial characterization of a Paracoccidioides brasiliensis protein with capacity to bind to extracellular matrix proteins. AB - Microorganisms adhere to extracellular matrix proteins by means of their own surface molecules. Paracoccidioides brasiliensis conidia have been shown to be capable of interacting with extracellular matrix proteins. We aimed at determining the presence of fungal proteins that could interact with extracellular matrix protein and, if found, attempt their purification and characterization. Various extracts were prepared from P. brasiliensis mycelial and yeast cultures (total homogenates, beta-mercaptoethanol, and sodium dodecyl sulfate [SDS] extracts) and analyzed by ligand affinity assays with fibronectin, fibrinogen and laminin. Two polypeptides were detected in both fungal forms. SDS extracts that interacted with all the extracellular matrix protein were tested; their molecular masses were 19 and 32 kDa. Analysis of the N-terminal amino acid sequence of the purified 32-kDa mycelial protein showed substantial homology with P. brasiliensis, Histoplasma capsulatum, and Neurospora crassa hypothetical proteins. Additionally, a monoclonal antibody (MAb) produced against this protein recognized the 32-kDa protein in the SDS extracts of both fungal forms for immunoblot. Immunofluorescence analysis revealed that this MAb reacted not only with mycelia and yeast cells, but also with conidia, indicating that this protein was shared by the three fungal propagules. By immunoelectron microscopy, this protein was detected in the cell walls and in the cytoplasm. Both the 32-kDa purified protein and MAb inhibited the adherence of conidia to the three extracellular matrix proteins in a dose-dependent manner. These findings demonstrate the presence of two polypeptides capable of interacting with extracellular matrix proteins on the surface of P. brasiliensis propagules, indicating that there may be common receptors for laminin, fibronectin, and fibrinogen. These proteins would be crucial for initial conidial adherence and perhaps also in dissemination of paracoccidioidomycosis. PMID- 15784596 TI - Role of endothelin 1 in the pathogenesis of chronic chagasic heart disease. AB - On the basis of previous observations, endothelin 1 (ET-1) has been suggested as contributing to the pathogenesis of Chagasic cardiomyopathy. Therefore, ET 1flox/flox;alpha-MHC-Cre(+) mice in which the ET-1 gene was deleted from cardiac myocytes and ET-1flox/flox;Tie 2 Cre(+) mice in which the ET-1 gene was deleted from endothelial cells were infected with Trypanosoma cruzi. Genetic controls for these cell-specific ET-1 knockout mice were used. Ninety percentage of all mice survived acute infection with the Brazil strain and were evaluated 130 days postinfection. Inflammation and fibrosis were observed in all infected mice; however, fibrosis was reduced in ET-1flox/flox;alpha-MHC-Cre(+) mice. Cardiac magnetic resonance imaging revealed that infection resulted in a significant increase in right ventricular internal diameter (RVID) in all mice except ET 1flox/flox;alpha-MHC-Cre(+) mice; i.e., RVID was not changed in infected ET 1flox/flox;alpha-MHC-Cre(+) mice. Echocardiography of the left ventricle demonstrated increased left ventricular end-diastolic diameter, reduced fractional shortening, and decreased relative wall thickness in infected mice. However, the magnitude of the changes was significantly less in ET 1flox/flox;alpha-MHC-Cre(+) mice compared to other groups. These data provide further evidence of a role for ET-1, particularly cardiac myocyte-derived ET-1, in the pathogenesis of chronic Chagasic cardiomyopathy. PMID- 15784597 TI - Novel mouse model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis. AB - Pseudomonas aeruginosa causes a chronic infection in the lungs of cystic fibrosis (CF) patients by establishing an alginate-containing biofilm. The infection has been studied in several animal models; however, most of the models required artificial embedding of the bacteria. We present here a new pulmonary mouse model without artificial embedding. The model is based on a stable mucoid CF sputum isolate (NH57388A) with hyperproduction of alginate due to a deletion in mucA and functional N-acylhomoserine lactone (AHL)-based quorum-sensing systems. Chronic lung infection could be established in both CF mice (Cftr(tmlUnc-/-)) and BALB/c mice, as reflected by the detection of a high number of P. aeruginosa organisms in the lung homogenates at 7 days postinfection and alginate biofilms, surrounded by polymorphonuclear leukocytes in the alveoli. In comparison, both an AHL producing nonmucoid revertant (NH57388C) from the mucoid isolate (NH57388A) and a nonmucoid isolate (NH57388B) deficient in AHL were almost cleared from the lungs of the mice. This model, in which P. aeruginosa is protected against the defense system of the lung by alginate, is similar to the clinical situation. Therefore, the mouse model provides an improved method for evaluating the interaction between mucoid P. aeruginosa, the host, and antibacterial therapy. PMID- 15784598 TI - Potential role of the chemokine macrophage inflammatory protein 1alpha in human and experimental schistosomiasis. AB - In human schistosomiasis, the concentrations of the chemokine macrophage inflammatory protein 1alpha (MIP-1alpha/CCL3) is greater in the plasma of patients with clinical hepatosplenic disease. The objective of the present study was to confirm the ability of CCL3 to detect severe disease in patients classified by ultrasonography (US) and to evaluate the potential role of CCL3 in Schistosoma mansoni-infected mice. CCL3 was measured by enzyme-linked immunosorbent assay in the plasma of S. mansoni-infected patients. CCL3-deficient mice were infected with 25 cercariae, and various inflammatory and infectious indices were evaluated. The concentration of CCL3 was higher in the plasma of S. mansoni-infected than noninfected patients. Moreover, CCL3 was greater in those with US-defined hepatosplenic than with the intestinal form of the disease. In CCL3-deficient mice, the size of the granuloma and the liver eosinophil peroxidase activity and collagen content were diminished compared to wild-type mice. In CCL3-deficient mice, the worm burden after 14 weeks of infection, but not after 9 weeks, was consistently smaller. The in vitro response of mesenteric lymph node cells to antigen stimulation was characterized by lower levels of interleukin-4 (IL-4) and IL-10. CCL3 is a marker of disease severity in infected humans, and experimental studies in mice suggest that CCL3 may be a causative factor in the development of severe schistosomiasis. PMID- 15784599 TI - Shiga toxin is transported from the endoplasmic reticulum following interaction with the luminal chaperone HEDJ/ERdj3. AB - Shiga toxin (Stx) follows a complex intracellular pathway in order to kill susceptible cells. After binding to cell surface glycolipids, the toxin is internalized and trafficked in retrograde fashion to the endoplasmic reticulum (ER). From the ER lumen, the toxin must gain access to the cytoplasm, where it enzymatically inactivates the 28S rRNA, inhibiting protein synthesis. The host molecules involved in this pathway and the mechanisms utilized by the toxin to access the cytoplasm from the ER are largely unknown. We found that Stx is capable of energy-dependent transport across the ER lumen, as has recently been demonstrated for the cholera and ricin toxins. Genetic screening for molecules involved in Shiga toxin trafficking yielded a cDNA encoding a prematurely truncated protein. Characterization of this cDNA revealed that it encodes a novel Hsp40 chaperone, designated HEDJ or ERdj3, localized to the ER lumen, where it interacts with BiP, a molecule known to be involved in protein retrotranslocation out of the ER. We demonstrated that within the ER lumen Stx interacts with HEDJ and other chaperones known to be involved in retrotranslocation of proteins across the ER membrane. Moreover, sequential immunoprecipitation revealed that Shiga toxin was present in a complex that included HEDJ and Sec61, the translocon through which proteins are retrotranslocated to the cytoplasm. These findings suggest that HEDJ is a component of the ER quality control system and that Stx utilizes HEDJ and other ER-localized chaperones for transport from the ER lumen to the cytosol. PMID- 15784600 TI - Designer arrays for defined mutant analysis to detect genes essential for survival of Mycobacterium tuberculosis in mouse lungs. AB - The mechanisms by which Mycobacterium tuberculosis elicits disease are complex, involving a large repertoire of bacterial genes that are required for in vivo growth and survival. To identify such genes, we utilized a high-throughput microarray detection method to rapidly screen hundreds of unique, genotypically defined transposon mutants for in vivo survival with a high degree of specificity and sensitivity. Thirty-one M. tuberculosis genes were found to be required for in vivo survival in mouse lungs. These genes are involved in a broad range of activities, including metabolism, cell wall functions, and regulation. Our screen included 11 of the 12 known members of the mycobacterial membrane protein (mmpL) family genes, and mutation of 6 of these genes-mmpL4, mmpL5, mmpL7, mmpL8, mmpL10, and mmpL11-severely compromised the ability of the mutants to multiply in mouse lungs. Most of the 31 genes are conserved in other pathogenic mycobacteria, including M. leprae and M. bovis, suggesting that a core of basic in vivo survival mechanisms may be highly conserved despite the divergent human pathology caused by members of the mycobacterial genus. Of the 31 genes reported here, 17 have not been previously described to be involved in in vivo growth and survival of M. tuberculosis. PMID- 15784602 TI - Isocitrate lyase (AceA) is required for Salmonella persistence but not for acute lethal infection in mice. AB - Isocitrate lyase is required for fatty acid utilization via the glyoxylate shunt. Although isocitrate lyase is essential for Salmonella persistence during chronic infection, it is dispensable for acute lethal infection in mice. Substrate availability in the phagosome appears to evolve over time, with increasing fatty acid dependence during chronic infection. PMID- 15784601 TI - Antibody against the carboxyl terminus of intimin alpha reduces enteropathogenic Escherichia coli adherence to tissue culture cells and subsequent induction of actin polymerization. AB - The C-terminal third of intimin binds to its translocated receptor (Tir) to promote attaching and effacing lesion formation during infection with enteropathogenic Escherichia coli (EPEC). We observed that the adherence of EPEC strains to HEp-2 cells was reduced and that actin polymerization was blocked by antibody raised against the C-terminal third of intimin alpha. PMID- 15784603 TI - Pseudomonas aeruginosa OxyR is required for full virulence in rodent and insect models of infection and for resistance to human neutrophils. PMID- 15784604 TI - The mannose receptor mediates the cellular immune response in human coccidioidomycosis. AB - Mannose is the predominant monosaccharide in the coccidioidal antigen preparation T27K. Mannan and anti-CD206 antibody significantly decreased the surface expression of mannose receptor (MR) on adherent peripheral blood mononuclear cells and reduced the interleukin-2 (IL-2) release induced by T27K. These data suggest that MR mediates IL-2 release by T27K. PMID- 15784605 TI - Linkage of the horizontally acquired ypm and pil genes in Yersinia pseudotuberculosis. AB - The superantigen-encoding ypm gene and the pil gene cluster governing type IV pilus biogenesis have been laterally acquired by Yersinia pseudotuberculosis. PCR assays on 270 unrelated strains from various environmental and animal sources revealed a significant association of ypm and pil in isolates. PMID- 15784606 TI - Nonopsonic phagocytosis of erythrocytes infected with ring-stage Plasmodium falciparum. AB - Ring-stage parasitized erythrocytes (RPEs) were demonstrated to interact with effector cells of the innate immune system. With receptor blockade studies and CD36-null macrophages, human and murine macrophages were shown to phagocytose RPEs through the pattern recognition receptor CD36. These in vitro data implicate scavenger receptors in the clearance of RPEs. PMID- 15784607 TI - Leishmania major modulates chemokine and chemokine receptor expression by dendritic cells and affects their migratory capacity. AB - Dendritic cells (DC) both produce and respond to chemokines. We examined the profiles of chemokines and chemokine receptors expressed by DC and their chemotactic response after interaction with Leishmania major. Expression of the chemokine receptors CCR2 and CCR5 by DC and their responsiveness to the respective ligands, CCL2 and CCL3, were downregulated, while the level of CCR7 and the DC response to its ligand CCL21 were enhanced. These parasite-induced alterations were observed with DC from L. major-resistant and -susceptible mice. In contrast, expression of the chemokine CXCL10 was elicited only in DC from L. major-resistant mice. PMID- 15784608 TI - Differences in levels of secreted locus of enterocyte effacement proteins between human disease-associated and bovine Escherichia coli O157. PMID- 15784609 TI - Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction. AB - Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline transactivator (rtTA)/tetracycline-responsive element (tet-O)-driven transgenes. To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist. PMID- 15784610 TI - Simple and straightforward construction of a mouse gene targeting vector using in vitro transposition reactions. AB - In a gene targeting experiment, the generation of a targeting construct often requires complex DNA manipulations. We developed a set of cassettes and plasmids useful for creating targeting vectors to modify the mammalian genome. A positive selection marker cassette (PGK/EM7p-npt), which included dual prokaryotic and eukaryotic promoters to permit consecutive selection for recombination in Escherichia coli and then in mouse embryonic stem cells, was flanked by two FRT loxP sequences. The PGK/EM7p-npt cassette was placed between the minimum regions of a Tn7 transposable element for insertion into another DNA by means of Tn7 transposase in vitro. We also constructed a plasmid having a loxP-Zeo-loxP cassette between the modified Tn5 outer elements. These cassettes can be integrated randomly into a given genomic DNA through the in vitro transposition reaction, thus producing a collection of genomic segments flanked by loxP sites (floxed) at various positions without the use of restriction enzymes and DNA ligase. We confirmed that this system remarkably reduced the time and labor for the construction of complex gene targeting vectors. PMID- 15784611 TI - Sequence variation in G-protein-coupled receptors: analysis of single nucleotide polymorphisms. AB - We assessed the disease-causing potential of single nucleotide polymorphisms (SNPs) based on a simple set of sequence-based features. We focused on SNPs from the dbSNP database in G-protein-coupled receptors (GPCRs), a large class of important transmembrane (TM) proteins. Apart from the location of the SNP in the protein, we evaluated the predictive power of three major classes of features to differentiate between disease-causing mutations and neutral changes: (i) properties derived from amino-acid scales, such as volume and hydrophobicity; (ii) position-specific phylogenetic features reflecting evolutionary conservation, such as normalized site entropy, residue frequency and SIFT score; and (iii) substitution-matrix scores, such as those derived from the BLOSUM62, GRANTHAM and PHAT matrices. We validated our approach using a control dataset consisting of known disease-causing mutations and neutral variations. Logistic regression analyses indicated that position-specific phylogenetic features that describe the conservation of an amino acid at a specific site are the best discriminators of disease mutations versus neutral variations, and integration of all our features improves discrimination power. Overall, we identify 115 SNPs in GPCRs from dbSNP that are likely to be associated with disease and thus are good candidates for genotyping in association studies. PMID- 15784615 TI - Base pair opening in three DNA-unwinding elements. AB - DNA-unwinding elements are specific base sequences that are located in the origin of DNA replication where they provide the start point for strand separation and unwinding of the DNA double helix. In the present work we have obtained the first characterization of the opening of individual base pairs in DNA-unwinding elements. The three DNA molecules investigated reproduce the 13-mer DNA-unwinding elements present in the Escherichia coli chromosome. The base sequences of the three 13-mers are conserved in the origins of replication of enteric bacterial chromosomes. The exchange of imino protons with solvent protons was measured for each DNA as a function of the concentration of exchange catalyst using nuclear magnetic resonance spectroscopy. The exchange rates provided the rates and the equilibrium constants for opening of individual base pairs in each DNA at 20 degrees C. The results reveal that the kinetics and energetics of the opening reactions for AT/TA base pairs are different in the three DNA-unwinding elements due to long range effects of the base sequence. These differences encompass the AT/TA base pairs that are conserved in various bacterial genomes. Furthermore, a qualitative correlation is observed between the kinetics and energetics of opening of AT/TA base pairs and the location of the corresponding DNA-unwinding element in the origin of DNA replication. PMID- 15784612 TI - Transcriptional regulation of two stage-specifically expressed genes in the protozoan parasite Toxoplasma gondii. AB - The protozoan parasite Toxoplasma gondii differentially expresses two distinct enolase isoenzymes known as ENO1 and ENO2, respectively. To understand differential gene expression during tachyzoite to bradyzoite conversion, we have characterized the two T.gondii enolase promoters. No homology could be found between these sequences and no TATA or CCAAT boxes were evident. The differential activation of the ENO1 and ENO2 promoters during tachyzoite to bradyzoite differentiation was investigated by deletion analysis of 5'-flanking regions fused to the chloramphenicol acetyltransferase reporter followed by transient transfection. Our data indicate that in proliferating tachyzoites, the repression of ENO1 involves a negative distal regulatory region (nucleotides -1245 to -625) in the promoter whereas a proximal regulatory region in the ENO2 promoter directs expression at a low level. In contrast, the promoter activity of ENO1 is highly induced following the conversion of tachyzoites into resting bradyzoites. The ENO2 promoter analysis in bradyzoites showed that there are two upstream repression sites (nucleotides -1929 to -1067 and -456 to -222). Furthermore, electrophoresis mobility shift assays demonstrated the presence of DNA-binding proteins in tachyzoite and bradyzoite nuclear lysates that bound to stress response elements (STRE), heat shock-like elements (HSE) and other cis-regulatory elements in the upstream regulatory regions of ENO1 and ENO2. Mutation of the consensus AGGGG sequence, completely abolished protein binding to an oligonucleotide containing this element. This study defines the first characterization of cis-regulatory elements and putative transcription factors involved in gene regulation of the important pathogen T.gondii. PMID- 15784616 TI - Coupled translation of the respiratory syncytial virus M2 open reading frames requires upstream sequences. AB - We have investigated the mechanism of the translation of the second open reading frame (ORF) of the respiratory syncytial virus M2 transcript that uses a novel coupled translation process requiring prior translation of the upstream ORF. The second M2-2 ORF sequences play no role in the coupling process and can be replaced with other gene sequences. Surprisingly, the overlap region of the two ORFs alone was not sufficient for coupled translation to occur. An analysis of the sequences required for the coupling process showed that portions of the transcript located along the length of the first ORF M2-1, upstream of the ORF overlap region, were essential for coupled translation to occur. A critically important region for this process was centered approximately 150 nucleotides upstream of the ORF2 initiation codons. This region was shown to contain a significant degree of secondary structure, and mutation of this sequence to remove predicted areas of base pairing significantly reduced coupled translation, confirming that the secondary structure was important for the coupling process. Additional sequences further upstream increased the efficiency of the coupled translation process. These data indicate that upstream sequences act in conjunction with the M2-1/M2-2 overlap region to promote coupled translation. PMID- 15784617 TI - NRIP, a novel nuclear receptor interaction protein, enhances the transcriptional activity of nuclear receptors. AB - Transcriptional regulation by members of the nuclear hormone receptor superfamily is a modular process requiring the mediation of distinct subclasses of coregulators. In this study, we isolated a novel WD40 repeat-containing gene, human nuclear receptor interaction protein (NRIP). We found NRIP interacts with either androgen or glucocorticoid receptors from in vitro and in vivo pulldown assays. Subsequently, transient transfection and luciferase activity assays suggested that NRIP was a ligand-dependent coactivator of steroid receptors (androgen and glucocorticoid) in distinct promoters. To further clarify the function of NRIP, we found an RNA interference-3-targeted NRIP gene sequence (5' GATGATACAGCACGAGAAC-3') that could efficiently and specifically knock down endogenous and exogenous NRIP gene expression and that significantly diminished cell proliferation in prostate (LNCaP) and cervical (C33A) cells. Therefore, NRIP may play a role in enhancing the transcriptional activity of nuclear receptors and may be a critical target for developing therapeutic agents against nuclear receptor-mediated progression of prostate and cervical cancers. PMID- 15784618 TI - Probing the mechanism of ligand recognition in family 29 carbohydrate-binding modules. AB - The recycling of photosynthetically fixed carbon, by the action of microbial plant cell wall hydrolases, is integral to one of the major geochemical cycles and is of considerable industrial importance. Non-catalytic carbohydrate-binding modules (CBMs) play a key role in this degradative process by targeting hydrolytic enzymes to their cognate substrate within the complex milieu of polysaccharides that comprise the plant cell wall. Family 29 CBMs have, thus far, only been found in an extracellular multienzyme plant cell wall-degrading complex from the anaerobic fungus Piromyces equi, where they exist as a CBM29-1:CBM29-2 tandem. Here we present both the structure of the CBM29-1 partner, at 1.5 A resolution, and examine the importance of hydrophobic stacking interactions as well as direct and solvent-mediated hydrogen bonds in the binding of CBM29-2 to different polysaccharides. CBM29 domains display unusual binding properties, exhibiting specificity for both beta-manno- and beta-gluco-configured ligands such as mannan, cellulose, and glucomannan. Mutagenesis reveals that "stacking" of tryptophan residues in the n and n+2 subsites plays a critical role in ligand binding, whereas the loss of tyrosine-mediated stacking in the n+4 subsite reduces, but does not abrogate, polysaccharide recognition. Direct hydrogen bonds to ligand, such as those provided by Arg-112 and Glu-78, play a pivotal role in the interaction with both mannan and cellulose, whereas removal of water-mediated interactions has comparatively little effect on carbohydrate binding. The interactions of CBM29-2 with the O2 of glucose or mannose contribute little to binding affinity, explaining why this CBM displays dual gluco/manno specificity. PMID- 15784619 TI - The crystal structure of the collagen-like polypeptide (glycyl-4(R)-hydroxyprolyl 4(R)-hydroxyprolyl)9 at 1.55 A resolution shows up-puckering of the proline ring in the Xaa position. AB - The collagen triple helix is characterized by the repeating sequence motif Gly Xaa-Yaa, where Xaa and Yaa are typically proline and (2S,4R)-4-hydroxyproline (4(R)Hyp), respectively. Previous analyses have revealed that H-(Pro-4(R)Hyp Gly)(10)-OH forms a stable triple helix, whereas H-(4(R)Hyp-Pro-Gly)(10)-OH does not. Several theories have been put forth to explain the importance of proline puckering and conformation in triple helix formation; however, the details of how they affect triple helix stability are unknown. Underscoring this, we recently demonstrated that the polypeptide Ac-(Gly-4(R)Hyp-4(R)Hyp)(10)-NH(2) forms a triple helix that is more stable than Ac-(Gly-Pro-4(R)Hyp)(10)-NH(2). Here we report crystal the structure of the H-(Gly-4(R)Hyp-4(R)Hyp)(9)-OH peptide at 1.55 A resolution. The puckering of the Yaa position 4(R)Hyp in this structure is up (Cgamma exo), as has been found in other collagen peptide structures. Notably, however, the 4(R)Hyp in the Xaa position also takes the up pucker, which is distinct from all other collagen structures. Regardless of the notable difference in the Xaa proline puckering, our structure still adopts a 7/2 superhelical symmetry similar to that observed in other collagen structures. Thus, the basis for the observed differences in the thermodynamic data of the triple helix<--> coil transition between our peptide and other triple helical peptides likely results from contributions from the unfolded state. Indeed, the unfolded state of the H-(Gly-4(R)Hyp-4(R)Hyp)(9)-OH peptide seems to be stabilized by a preformed polyproline II helix in each strand, which could be explained by the presence of a unique repeating intra-strand water-mediated bridge observed in the H-(Gly 4(R)Hyp-4(R)Hyp)(9)-OH structure, as well as a higher amount of trans peptide bonds. PMID- 15784620 TI - Binding of iron-free siderophore, a common feature of siderophore outer membrane transporters of Escherichia coli and Pseudomonas aeruginosa. AB - TonB-dependent iron transporters present in the outer membranes of Gram-negative bacteria transport ferric-siderophore complexes into the periplasm. This requires proton motive force and an integral inner membrane complex, TonB-ExbB-ExbD. Recognition of iron-free siderophores by TonB-dependent outer membrane transporters (OMT) has only been described for a subfamily called OMT(N). These OMT(N)s have an additional domain at the N terminus, which interacts with an inner membrane regulatory protein to activate a cytoplasmic sigma factor. This induces transcription of iron transport genes. Here we showed that the ability to bind aposiderophores is not specific to the OMT(N) subfamily but may be a more general feature of OMTs. FhuA, the ferrichrome OMT in Escherichia coli, and FptA, the pyochelin (Pch) OMT in Pseudomonas aeruginosa, were both able to bind in vitro and in vivo the apo-forms and the ferric forms of their corresponding siderophore at a common binding site. FptA produced in P. aeruginosa cells grown in an iron-deficient medium copurifies with a ligand that, as characterized by fluorescence, is iron-free Pch. As described previously for the FpvA transporter (pyoverdine OMT in P. aeruginosa), it appears that in conditions of iron limitation all the FptA receptors at the cell surface are loaded with apoPch. This FptA-Pch complex is less stable in vitro than the previously described copurified FpvA-Pvd complex and can be loaded with iron in vitro in the presence of Pch-Fe, citrate-Fe, or ferrichrome-Fe. These findings improved our understanding of the iron uptake mechanism via siderophores in Gram-negative bacteria. PMID- 15784621 TI - Regulation of MDR1 expression and drug resistance by a positive feedback loop involving hyaluronan, phosphoinositide 3-kinase, and ErbB2. AB - Multidrug resistance is a potent barrier to effective, long term therapy in cancer patients. It is frequently attributed to enhanced expression of multidrug transporters or to the action of receptor kinases, such as ErbB2, and downstream anti-apoptotic signaling pathways, such as the phosphoinositide 3-kinase/Akt pathway. However, very few connections have been made between receptor kinases or anti-apoptotic pathways and multidrug transporter expression or function. Data presented herein show that constitutive interaction of the pericellular polysaccharide, hyaluronan, with its receptor, CD44, regulates assembly and activation of an ErbB2-containing signaling complex, which in turn stimulates phosphoinositide 3-kinase activity in multidrug-resistant MCF-7/Adr human breast carcinoma cells. Phosphoinositide 3-kinase activates Akt and downstream anti apoptotic events, which contribute to drug resistance. However, hyaluronan and phosphoinositide 3-kinase stimulate expression of the multidrug transporter, MDR1 (P-glycoprotein), in an interdependent, but Akt-independent, manner. Furthermore, constitutively active phosphoinositide 3-kinase, but not Akt, stimulates hyaluronan production. These Akt-independent effects are dominant over the effects of Akt on doxorubicin resistance in MCF-7/Adr cells. Thus hyaluronan, phosphoinositide 3-kinase, and ErbB2 form a positive feedback loop that strongly amplifies MDR1 expression and regulates drug resistance in these cells. This pathway may also be important in progression of other malignant characteristics. These results illustrate the potential importance of hyaluronan as a therapeutic target in multidrug-resistant carcinomas. PMID- 15784622 TI - ArgBP2gamma interacts with Akt and p21-activated kinase-1 and promotes cell survival. AB - Akt/protein kinase B is a major cell survival pathway through phosphorylation of proapoptotic proteins Bad and Bax and of additional apoptotic pathways linked to Forkhead proteins glycogen synthase kinase-3beta and ASK1. To further explore the mechanism by which Akt regulates cell survival, we identified an Akt interaction protein by yeast two-hybrid screening. It is highly homologous to ARG-binding protein 2 (ArgBP2) with splicing exon 8 of the coding region of the ArgBP2. As two splicing isoforms (ArgBP2alpha and -beta) of ArgBP2 have been identified (Wang, B., Golemis, E. A., and Kruh, G. D. (1997) J. Biol. Chem. 272, 17542 17550), it was named ArgBP2gamma. ArgBP2gamma contains four Akt phosphorylation consensus sites, a SoHo motif, and three Src homology (SH) 3 domains and binds to C-terminal proline-rich motifs of Akt through its first and second SH3 domains. It also interacts with p21-activated protein kinase (PAK1) via its first and third SH3 domains, indicating the SH3 domains of ArgBP2gamma as docking sites for Akt and PAK1. Akt phosphorylates ArgBP2gamma in vitro and in vivo. Expression of ArgBP2gamma induces PAK1 activity and overrides apoptosis induced by ectopic expression of Bad or DNA damage. Nonphosphorylatable ArgBP2gamma-4A and SH3 domain-truncated mutant ArgBP2gamma inhibit Akt-induced PAK1 activation and reduce Akt and PAK1 phosphorylation of Bad and antiapoptotic function. These data indicate that ArgBP2gamma is a physiological substrate of Akt, functions as an adaptor for Akt and PAK1, and plays a role in Akt/PAK1 cell survival pathway. PMID- 15784623 TI - Transporter-associated currents in the gamma-aminobutyric acid transporter GAT-1 are conditionally impaired by mutations of a conserved glycine residue. AB - To determine whether glycine residues play a role in the conformational changes during neurotransmitter transport, we have analyzed site-directed mutants of the gamma-aminobutyric acid (GABA) transporter GAT-1 in a domain containing three consecutive glycines conserved throughout the sodium- and chloride-dependent neurotransmitter transporter family. Only cysteine replacement of glycine 80 resulted in the complete loss of [(3)H]GABA uptake, but oocytes expressing this mutant exhibited the sodium-dependent transient currents thought to reflect a charge-moving conformational change. When sodium was removed and subsequently added back, the transients by G80C did not recover, as opposed to wild type, where recovery was almost complete. Remarkably, the transients by G80C could be restored after exposure of the oocytes to either GABA or a depolarizing pre pulse. These treatments also resulted in a full recovery of the transients by the wild type. Whereas in wild type lithium leak currents are observed after prior sodium depletion, this was not the case for the glycine 80 mutants unless GABA was added or the oocytes were subjected to a depolarizing pre-pulse. Thus, glycine 80 appears essential for conformational transitions in GAT-1. When this residue is mutated, removal of sodium results in "freezing" the transporter in one conformation from which it can only exit by compensatory changes induced by GABA or depolarization. Our results can be explained by a model invoking two outward-facing states of the empty transporter and a defective transition between these states in the glycine 80 mutants. PMID- 15784625 TI - Nectin-4, a new serological breast cancer marker, is a substrate for tumor necrosis factor-alpha-converting enzyme (TACE)/ADAM-17. AB - Serum markers are extensively used in diagnostic and follow-up of cancer patients. We recently described Nectin-4, a 66-kDa adhesion molecule of the Nectin family, which is a valuable new histological and serological marker for breast carcinoma. In vivo, Nectin-4 is re-expressed in breast carcinoma, and a circulating form of Nectin-4 is detected in the sera of patients with metastatic breast cancer. In vitro, a soluble form of Nectin-4 is produced in the supernatant of breast tumor cell lines (S. Fabre-Lafay, C. Ginestier, S. Garrido Urbani, C. Berruyer, R. Sauvan, N. Reymond, J. Adelaide, J. Geneix, P. Dubreuil, J. Jacquemier, D. Birnbaum, and M. Lopez, manuscript in preparation). We have investigated the mechanisms that regulate the production of this soluble form. It was found that the soluble form of Nectin-4 detected in the sera of patients and the supernatant of breast tumor cell lines share similar biochemical and immunological features. The soluble Nectin-4 form (43 kDa) is formed by the entire Nectin-4 ectodomain. Nectin-4 shedding is constitutive, strongly enhanced by 12-O-tetradecanoylphorbol-13-acetate activation, and reduced tumor necrosis factor-alpha protease inhibitor TAPI-1 or by the tissue inhibitor of metalloproteinase-3 (TIMP-3). TAPI-1 and TIMP-3 are inhibitors of the endoprotease tumor necrosis factor-alpha-converting enzyme (TACE)/ADAM-17. Overexpression or small interfering RNA-mediated silencing of TACE enhanced or reduced Nectin-4 shedding, respectively. Nectin-4 is not shed when expressed in TACE-deficient fibroblasts. Interestingly, the active form of TACE is overexpressed in breast tumors and may indicate that TACE is responsible for Nectin-4 shedding not only in vitro but also in vivo. PMID- 15784624 TI - Chemical rescue of histidine selectivity filter mutants of the M2 ion channel of influenza A virus. AB - The influenza virus M2 proton-selective ion channel activity facilitates virus uncoating, a process that occurs in the acidic environment of the endosome. The M2 channel causes acidification of the interior of the virus particle, which results in viral protein-protein dissociation. The M2 protein is a homotetramer that contains in its aqueous pore a histidine residue (His-37) that acts as a selectivity filter and a tryptophan residue (Trp-41) that acts as a channel gate. Substitution of His-37 modifies M2 ion channel properties drastically. However, the results of such experiments are difficult to interpret because substitution of His-37 could cause gross structural changes to the channel pore. We described here experiments in which partial or, in some cases, full rescue of specific M2 ion channel properties of His-37 substitution mutants was achieved by addition of imidazole to the bathing medium. Chemical rescue was demonstrated for three histidine substitution mutant ion channels (M2-H37G, M2-H37S, and M2-H37T) and for two double mutants in which the Trp-41 channel gate was also mutated (H37G/W41Y and H37G/W41A). Currents of the M2-H37G mutant ion channel were inhibited by Cu(II), which has been shown to coordinate with His-37 in the wild type channel. Chemical rescue was very specific for imidazole. Buffer molecules that were neutral when protonated (4-morpholineethanesulfonic acid and 3 morpholino-2-hydroxypropanesulfonic acid) did not rescue ion channel activity of the M2-H37G mutant ion channel, but 1-methylimidazole did provide partial rescue of function. These results were consistent with a model for proton transport through the pore of the wild-type channel in which the imidazole side chain of His-37 acted as an intermediate proton acceptor/donor group. PMID- 15784626 TI - Endogenous calcitonin gene-related peptide protects human alveolar epithelial cells through protein kinase Cepsilon and heat shock protein. AB - The intracellular mechanisms of ischemic preconditioning (PC) in preventing lung dysfunction following transplantation, shock, and trauma remain poorly understood. Previously, we have shown that alveolar epithelial cells secrete calcitonin gene-related peptide (CGRP) under inflammatory stress. Using a hypoxia/reoxygenation (H/R) and PC model, we found that CGRP was also secreted from human type II alveolar epithelial cells (A549) after PC. The locally released CGRP interacted with its receptor on the membrane of A549 cells and elicited downstream signals mediating the PC effect, because hCGRP(8-37), a specific CGRP receptor antagonist, attenuated the protective effect of PC. Pre inhibition of CGRP protein synthesis by small interfering RNA exacerbated (but overexpression of the CGRP gene ameliorated) H/R-induced cell death, which supports the autocrine effect of CGRP on A549 cells. Exogenous bioactive CGRP mimicked the beneficial effect of PC and up-regulated the expression of heat shock protein 70 (HSP70), which might act as the end effector to maintain cell viability. These effects were sensitive to hCGRP(8-37), calphostin C (a protein kinase C (PKC) inhibitor), and 5-hydroxydecanoic acid (a mitochondrial K(+)(ATP) channel blocker) but were insensitive to protein kinase A blockers. Moreover, CGRP induced the membrane translocation of PKCepsilon. PKCV1-2 (a cell-permeable inhibitory peptide of PKCepsilon) effectively abolished CGRP-induced HSP70 expression and cell protection. Therefore, PC induces CGRP secretion from human alveolar epithelial cells, and the locally released CGRP acts back on these cells, protecting them from H/R injury. The post-receptor signaling of CGRP is through PKCepsilon-dependent expression of HSP70. PMID- 15784627 TI - Anti-rheumatic drug use and risk of hospitalization for congestive heart failure in rheumatoid arthritis. AB - OBJECTIVE: To assess the risk of hospitalization for congestive heart failure (CHF) associated with the use of disease-modifying anti-rheumatic drugs (DMARDs) and other medications used in RA. METHODS: We used a case-control design nested within an administrative database cohort of patients with rheumatoid arthritis (RA) who were dispensed a DMARD between September 1998 and December 2001. Subjects identified with a prior history of CHF were excluded. For each hospitalized case of CHF identified during follow-up, 10 controls matched on age and time were randomly selected from the cohort. Conditional logistic regression was used to estimate the rate ratio (RR) of hospitalizations for CHF associated with the current use of specific drugs, adjusted for sex and co-morbidity. RESULTS: The cohort included 41 885 patients; 75% were women, with an average age at cohort entry of 51 yr. During follow-up, 520 hospitalizations for CHF occurred, for a rate of 10.1 per 1000 per year. The adjusted RR of CHF for current use of any DMARD was 0.7 (95% CI 0.6-0.9) relative to no current use. By DMARD category, there was evidence of a beneficial effect for both tumour necrosis factor-alpha antagonists (RR 0.5, 95% CI 0.2-0.9) and methotrexate monotherapy (RR 0.8, 95% CI 0.6-1.0). For non-DMARD medications, the rate of CHF was not clearly increased or decreased, except for COX-2 inhibitors. The data suggested an increased risk of CHF with rofecoxib (RR 1.3, 95% CI 1.0-3.1) and a decreased risk of CHF with celecoxib (RR 0.6, 95% CI 0.4, 1.0). CONCLUSIONS: The use of DMARDs was associated with a reduction in the risk of hospitalizations for CHF in this RA cohort. The increased risk with rofecoxib alongside a decreased risk with celecoxib suggests the absence of a class effect with respect to COX-II inhibitors for some types of cardiovascular morbidity. PMID- 15784628 TI - Increased stiffness of the abdominal aorta in women with rheumatoid arthritis. AB - OBJECTIVE: To study the distensibility and the diameter of the abdominal aorta and the common carotid artery (CCA) in patients with rheumatoid arthritis (RA), and investigate the relation between mechanical properties of these arteries and disease severity. METHODS: One hundred and one patients with RA (33 consecutive cases with extra-articular manifestations, and 68 subjects with non-extra articular disease, matched for age, sex and disease duration) were investigated. Echo-tracking ultrasonography was used to measure stiffness and mean diameter of the abdominal aorta and the CCA. The patients were compared with healthy individuals from the corresponding age group (n=74 for measurements of the aorta, n=64 for the CCA). Predicted values for stiffness and mean diameter, based on age and sex, were calculated. RESULTS: Stiffness of the abdominal aorta was increased in women with RA [mean percentage of predicted value (% predicted) 180; 95% confidence interval (95% CI) 150-211] but not in men (% predicted 99; 95% CI 75 122). CCA stiffness was less markedly increased, and mean diameters of the aorta and the CCA were not different from the expected. In the RA cohort, patients with extra-articular manifestations tended to have greater stiffness of the aorta (P=0.11), and disability, as indicated by a higher Health Assessment Questionnaire score, was associated with increased aortic stiffness (P=0.04). CONCLUSION: RA is associated with decreased distensibility of the abdominal aorta in females, and such changes seem to correlate with disease severity. We suggest that arterial stiffness is an important factor in cardiovascular co-morbidity in RA. PMID- 15784629 TI - Mycobacterium tuberculosis as a cause of chronic periaortitis. PMID- 15784630 TI - Is musculoskeletal pain more common now than 40 years ago?: Two population-based cross-sectional studies. AB - OBJECTIVE: To test the hypothesis that the prevalence of specific musculoskeletal pain symptoms has increased over time in the northwest region of England. To meet this objective we have examined the difference in the prevalence of low back, shoulder and widespread pain between the 1950s and today using historical data collected by the Arthritis Research Campaign (arc). METHODS: Two cross-sectional surveys conducted over 40 yr apart in the northwest region of England. The status of two regional pain sites and widespread pain was determined using interview and questionnaire responses, for the earlier and later studies respectively. Subjects were classified positively if they reported low back pain, shoulder pain or widespread pain on the day of the survey. Rates were standardized to the Greater Manchester population. RESULTS: There were large differences in the prevalence of musculoskeletal pain between the two surveys. For all three symptoms examined prevalence increased from 2- to 4-fold between the two surveys. In both surveys low back pain was more common in women. Shoulder and widespread pain was less prevalent in women than in men in the earlier survey but by the time of the later survey women reported more pain at these sites. CONCLUSIONS: The prevalence of musculoskeletal pain is much higher than that reported over 40 yr ago. The change in prevalence is unlikely to be entirely due to the study design; other possible explanations such as the increased reporting or awareness of these symptoms is discussed. PMID- 15784631 TI - What does "active disease" mean? patient and parent perceptions of disease activity in the systemic arthritis form of juvenile idiopathic arthritis (SO JIA). AB - OBJECTIVE: The systemic onset form of juvenile idiopathic arthritis (SO-JIA) is a very serious chronic illness of childhood. At present, there is no specific tool to measure disease activity for SO-JIA. Our long-term goal is to develop a disease activity measure for SO-JIA using a consensus (Delphi) approach. In preparation for the development of this measure, we interviewed both patients and their parents. We sought to elicit specific items reflecting their perceptions of active disease that may be considered for inclusion in a disease activity measure for SO-JIA. METHODS: SO-JIA patients followed at The Hospital for Sick Children and their parents were chosen by purposive sampling. A trained research coordinator interviewed all participants using open-ended questions to elicit aspects of disease activity (defined as reversible manifestations of disease) of relevance to families. A list of these aspects was then generated and organized by categories to allow item reduction. RESULTS: Fourteen patients (eight females) with a mean age of 11.8 yr (mean disease duration 5.2 yr) and their parents were interviewed. A total of 292 items were generated, with an average of 11 items generated per interview. Arthralgia, ambulation difficulties, rash, decreased energy level and fever were the most common items mentioned by patients and their parents. Mood disturbances, decreased activity levels, arthritis severity and decreased school attendance were also common items identified as relevant aspects of disease activity. CONCLUSION: This study has allowed us to include patient and parent perspectives in preparation for developing a disease activity measure for SO-JIA. The resulting items will be added to future physician surveys in the further development and validation of a disease activity measure for SO-JIA. PMID- 15784632 TI - Applications of biological control in resistant host-pathogen systems. AB - Insect pest species can have devastating effects on crops. Control of these insect pests is usually achieved by using chemical insecticides. However, there has been much cause for concern with their overuse. Consequently, research has been carried out into alternative forms of control, in particular biological control methods. Recent laboratory studies have indicated that these natural forms of control can induce resistant strains of insect pest. In this paper we present a discrete-time host-pathogen model to describe the interaction between a host (insect species) that can develop a resistant strain and a pathogen (biological control) that can be externally applied to the system. For this model we use a single-state variable for the host population. We show that the proportion of resistance in the population impacts on the viability of the host population. Moreover, when the host population does persist, we explore the interaction between host susceptibility and host population levels. The different scenarios which arise are explained ecologically in terms of trade-offs in intrinsic growth rates, disease susceptibility and intraspecific host competition for the resistant subclass. PMID- 15784633 TI - Risk factors for out-of-hospital cardiac arrest: the Reykjavik Study. AB - AIMS: To examine risk factors for out-of-hospital cardiac arrest in the Reykjavik Study, a long-term, prospective, population-based cohort study that started in 1967. METHODS AND RESULTS: From 1987 to 1996, 137 men and 44 women out of the 8006 men and 9435 women in the study sustained out-of-hospital cardiac arrest due to cardiac causes. Determinants included coronary artery disease (CAD), its classical risk factors, and age, body mass index (BMI), heart rate, cardiomegaly, and erythrocyte sedimentation rate. Electrocardiograms (ECGs) were examined for various abnormalities. Significance was determined by Cox regression analysis. In multivariable analysis, the risk in men was significantly associated with age, diastolic blood pressure, cholesterol, current smoking, and previous diagnosis of myocardial infarction (MI). In women, the risk was associated with diastolic blood pressure, elevated levels of cholesterol and triglycerides, and increased voltage on ECG. Increased BMI was inversely related to women's risk of out-of hospital cardiac arrest. CONCLUSION: In this prospective, population-based cohort study previous MI and the classical risk factors for CAD significantly increased the risk of out-of-hospital cardiac arrest, the endpoint of this study. Increased voltage on ECG additionally increased women's risk. PMID- 15784634 TI - Chylous ascites and chylothorax due to the existence of transdiaphragmatic shunting in an adult with nephrotic syndrome. PMID- 15784635 TI - Fatal renal and hepatic failure following silver nitrate instillation for treatment of chyluria. PMID- 15784636 TI - Penile gangrene in a chronic dialysis patient. PMID- 15784638 TI - Oral alkalinizing solution as a potential prophylaxis against myoglobinuric acute renal failure: preliminary data from healthy volunteers. AB - BACKGROUND: Acute renal failure (ARF) secondary to crush injury is one of the leading causes of hospitalization and death in survivors of massive disasters. The standard therapy for crush injury, intravenous (i.v.) hydration and alkalinization of urine, is often not feasible after a mass disaster; therefore, oral rehydration and urinary alkalinization may be a useful substitute. METHODS: We developed and evaluated an oral alkalinizing solution (OAS) to induce alkaline diuresis. We enrolled 12 volunteer Iranian Army recruits (mean age 19.4+/-0.8 years) who drank an average of 650 ml of OAS for 12 h. We checked the volume and pH of their urine every hour, and measured venous blood gas and electrolytes at 6, 12 and 15 h. RESULTS: All subjects tolerated the OAS without adverse events, and had active diuresis (>200 ml/h) after an average of 3.0+/-0.7 h. Their urine became alkaline (pH>7.0) within an average of 3.25+/-0.8 h. There were no significant electrolyte abnormalities. CONCLUSIONS: OAS seems to be a safe and promising means of inducing alkaline diuresis. It may be a feasible alternative to i.v. hydration to prevent ARF secondary to crush injuries in the context of mass disasters where i.v. hydration is not possible. A dose of 10 ml/kg/h may be the correct amount to induce alkaline diuresis within the first 12 h after crush injuries. The use of OAS for this purpose should be evaluated further. PMID- 15784637 TI - Novel therapeutic option in hypertensive crisis: sildenafil augments nitroprusside-induced hypotension. PMID- 15784639 TI - Pre-dilution on-line haemofiltration vs low-flux haemodialysis: a randomized prospective study. AB - BACKGROUND: Accumulation of larger molecular weight uraemic toxins molecules may have a negative effect on the cardiovascular and nutritional state of dialysis patients and influence uraemic symptomatology. Their clearance can be enhanced by the use of haemofiltration (HF). METHODS: The effects of low-flux haemodialysis (HD) (ultrapure dialysate; polyamide membranes) and pre-dilution on-line HF (1:1 blood/substitution ratio; target filtration volume: 1.2 times body weight) on cardiovascular and nutritional parameters, interdialytic levels of uraemic toxins and quality of life (QOL; Laupacis questionnaire) were assessed during 1 year follow-up. Forty patients were randomized. RESULTS: After 1 year, 27 patients were eligible for analysis (HF: 13 patients; HD: 14 patients). Left ventricular mass index did not change in the HF patients (127+/-33 --> 131+/-36 g/m(2) after 12 months) or in the HD group (135+/-34 --> 138+/-32 g/m(2)). Also, there were no changes in pulse wave velocity, and 48 h systolic and diastolic blood pressures. Lean body mass, assessed by dual-energy X-ray absorptiometry, increased in the HF group (44.8+/-8.9 --> 46.2+/-9.6 kg; P<0.05), but not in the HD group (49.4+/-9.2 --> 50.6+/-8.8 kg), although differences between groups were not significant. Insulin-like growth factor-1 levels remained stable in the HF patients, but decreased in the HD group (P<0.05 between groups). QOL for physical symptoms improved in the HF group (4.2+/-1.2 --> 5.0+/-1.1; P<0.05 within the HF group and P = 0.06 between groups), but not in the HD group (4.0+/-1.0 --> 4.4+/-1.4). beta2-microglobulin, complement factor D and homocysteine decreased significantly in the HF but not in the HD group, whereas l-ADMA, leptin and advanced glycation end-products-related fluorescence did not change. CONCLUSIONS: No changes in cardiovascular parameters were observed during pre-dilution on-line HF compared with low-flux HD. Treatment with on-line HF resulted in marked changes in the uraemic toxicity profile, an improvement in physical well-being and a small improvement in nutritional state. PMID- 15784640 TI - Peritoneal dialysis patient survival: a comparison between a Swedish and a Korean centre. AB - BACKGROUND: Dialysis patient mortality remains high, and this high mortality may be due to many factors. In peritoneal dialysis (PD) patients, old age, co-morbid diseases, malnutrition, low residual renal function (RRF) and a high peritoneal transport rate have been shown to influence survival, but the relative importance of these factors may differ between different patient populations. Besides, centre practice patterns may differ between centres and may influence patient survival. In addition, the literature suggests that dialysis patient survival may be better in Asian than in Caucasian patients. METHODS: The influence of centre and patient characteristics on patient survival was investigated in 132 Korean and 106 Swedish incident PD patients, who underwent initial biochemical measurements and assessment of adequacy of dialysis, nutritional status, RRF and peritoneal transport characteristics. RESULTS: At the start of PD, Korean patients had a higher prevalence of diabetes, peritoneal Kt/V(urea), peritoneal creatinine clearance and peritoneal fluid removal, and lower body mass index, RRF and dialysate to plasma creatinine concentration ratio (D/P Cr) compared with Swedish patients. Significantly more patients from Korea were placed on temporary haemodialysis before PD (100 out of 132) when compared with Swedish patients (21 out of 106). During the follow-up, there was a significantly higher rate of transfer to other units in Korea and a significantly higher rate of kidney transplantation in Sweden. On Kaplan-Meier analysis, overall patient survival did not differ and relative risk for death was also not different between the two centres even after adjustment for age, diabetes, cardiovascular disease, RRF and D/P Cr. On Cox proportional hazards multivariate analysis, age, diabetes, RRF and D/P Cr were found to be independent predictors of mortality in the combined cohort of patients. While age, diabetes and D/P Cr were independent predictors of mortality in Korean patients, age and RRF independently predicted mortality in Swedish patients. CONCLUSION: Although there were significant differences in centre and patient characteristics, we were unable to confirm a survival advantage for Korean over Swedish PD patients. The results of this study suggest that the reported difference in survival between Asian and Caucasian dialysis patients may have been due, in part, to differences in centre and patient characteristics rather than to race as such. The genetic influence on patient characteristics remains, however, to be elucidated. PMID- 15784641 TI - The effect of two different protocols of potassium haemodiafiltration on QT dispersion. AB - BACKGROUND: The risk of developing cardiovascular diseases is higher in patients on haemodialysis than in the general population. These patients may develop arrhythmias that depend on the extra- and intracellular concentrations of potassium. ECG findings, particularly the QT interval and its dispersion (QT(d)) and the QT(c) (QT interval corrected for heart rate according to Bazett's formula) and its dispersion (QT(cd)), may be direct indicators of the risk of developing arrhythmia. METHODS: Our cohort comprised 28 patients who were dialysed for 3.5-4 h three times per week, first with haemodiafiltration with a constant potassium concentration (HDF) in the dialysis bath then with haemodiafiltration with variable concentrations of potassium (HDF(k)). ECGs were done at different time intervals: at the start of dialysis (T(0)), at 15 (T(15)), 45 (T(45)), 90 (T(90)) and 120 min (T(120)) after the beginning of the session, and at the end of treatment (T(end)). ECG-derived data (QT, QT(d), QT(c) and QT(cd)) were measured. At the same time points, plasma electrolytes, intra erythrocytic potassium and the electrical membrane potential at rest (REMP) of the erythrocytic membrane were measured. RESULTS: Plasma potassium concentration diminished more gradually in HDF(k) than in HDF, the difference being statistically significant at T(15) and T(45) (P<0.05), and T(90) (P<0.01). The intra-erythrocytic potassium concentration remained constant throughout the observation period. In both HDF and HDF(k), REMP was lower at all points after T(0) (P<0.05), but the reduction was greater and more significant in HDF than in HDF(k) at T(15) and T(120) (P<0.05). ECG revealed a statistically significant diminution in HDF(k) vs HDF in the measures of dispersion of QT and QT(c) at T(15), T(90), T(120) and T(end) (P<0.01) and of QT(cd) at T(45) (P<0.05). The mean of QT(d), adjusted for plasma potassium, increased over time in HDF with large alternate mean increase and decrease peaks and error intervals. In HDF(k), instead, there was a progressive and constant diminution with minor error intervals. QT(cd) adjusted for plasma potassium had the same trend. A marked difference was found between the final values in standard HDF and those in HDF(k). CONCLUSIONS: HDF and HDF(k) have significantly different effects on QT(c). ECG data demonstrate that the risk of arrhythmia could be lower, with a variable removal of potassium during haemodialysis. With HDF but not HDF(k), hyperpolarization of the cell membrane is detected, and this could have a destabilizing effect on different types of cardiac cell, giving rise to retrograde circuits. PMID- 15784642 TI - Diagnosis and monitoring a case of light-chain deposition disease in the kidney using a new, sensitive immunoassay. AB - A 59-year-old male was diagnosed with nephrotic syndrome secondary to light-chain deposition disease. There was no other evidence of a B cell clonal disorder or amyloidosis; circulating free light chains were identified using a new immunoassay (Freelite) and used to monitor disease progression. Improvement in renal function and proteinuria following VAMP chemotherapy correlated with a reduction in circulating light-chain levels. This case demonstrates a new tool in monitoring light-chain deposition disease in the kidney. PMID- 15784643 TI - Hepatocyte growth factor and its receptor Met are induced in crescentic glomerulonephritis. AB - BACKGROUND: In experimental extracapillary glomerulonephritis (EG) podocytes migrate, proliferate and change phenotype, and play a pivotal role in crescent formation. Hepatocyte Growth Factor (HGF) is an injury-induced effector of tissue repair that causes cell migration, growth and transdifferentiation via its receptor Met. METHODS: In 11 patients with EG we measured serum levels of HGF and investigated whether serum induces the release of HGF by Peripheral Blood Mononuclear Cells (PBMC). In renal biopsies we studied the expression of Met. In cultured podocytes we studied Met expression, migration, growth and morphological changes induced by recombinant (r) HGF. RESULTS: In patients with EG average serum levels of HGF (0.73 ng/ml) were higher than in normal volunteers (N, 0.10 ng/ml, p<0.01) and in patients with non-crescentic glomerular disease (GD, 0.18 ng/ml, p<0.01). Serum of EG induced a significant HGF release by PBMC (mean 0.58 ng/ml) in comparison with serum of N and GD (0.07 and 0.06 ng/ml, respectively, both p<0.001). Met was strongly expressed in crescents. Cultured podocytes expressed Met, and rHGF induced in podocytes a time- and dose-dependent migration, growth and epithelial to mesenchymal transdifferentiation. CONCLUSIONS: These results suggest that HGF/Met system participates in the process of crescent formation by inducing podocyte migration, growth and mesenchymal transformation. PMID- 15784644 TI - Beneficial effect of early initiation of lipid-lowering therapy following renal transplantation. AB - BACKGROUND: Renal transplant recipients have a significantly reduced life expectancy, largely due to premature cardiovascular disease. The aim of the current analysis was to investigate the importance of time of initiation of therapy after transplantation, on the benefits of statin therapy. METHODS: 2102 renal transplant recipients with total cholesterol levels of 4.0-9.0 mmol/l were randomly assigned to treatment with fluvastatin (n = 1050) or placebo (n = 1052) and followed for a mean time of 5.1 years. The end-points were major cardiac events. The average median time from transplantation to randomization was 4.5 years (range: 0.5-29 years). RESULTS: In patients starting treatment with fluvastatin <4.5 years after renal transplantation, the incidence of cardiac events was 4.6% over 5.1 years vs 9.2% in those on placebo (P = 0.007). Fluvastatin significantly reduced the risk of cardiac death and non-fatal myocardial infarction by 56% [risk ratio (RR): 0.44; 95% confidence interval (95% CI): 0.26-0.74; P = 0.002]. In a more detailed analysis patients were grouped into 2-year intervals (since the last transplantation). The frequency of cardiac death and non-fatal myocardial infarction was reduced by 3.2%, 5.1%, 9.6% and 8.2% with fluvastatin treatment as compared to 6%, 10.4%, 13.4% and 9.6% with placebo when treatment was initiated at 0-2, 2-4, 4-6 and >6 years, respectively. The risk reduction for patients initiating therapy with fluvastatin at years 0-2 (compared with >6 years) following transplantation was 59% (RR: 0.41; 95% CI: 0.18-0.92; P = 0.0328). This is also reflected in total time on renal replacement therapy: in patients in the first quartile (<47 months) fluvastatin use was associated with a risk reduction of 64% compared with 19% for patients in the fourth quartile (>120 months) (P = 0.033). CONCLUSIONS: Our data support an early introduction of fluvastatin therapy in a population of transplant recipients at high risk of premature coronary heart disease. PMID- 15784645 TI - Effects of glycemic control on target organ responses to epinephrine in type 1 diabetes. AB - Severe hypoglycemia occurs in intensively treated patients with type 1 diabetes mellitus (T1DM) due in part to deficient epinephrine counterregulatory responses. Previously, we have found that T1DM patients demonstrated a spectrum of altered responses to epinephrine at a variety of target organs compared with nondiabetic healthy subjects. What is not known is whether intensive glycemic control further modifies target organ responses in individuals with T1DM. Therefore, the aim of this study is to assess whether there is tissue specific (liver, muscle, adipose tissue, pancreas and cardiovascular) resistance to epinephrine in intensively controlled (IC) T1DM compared with those with conventional control (CC). Eight IC patients (age 33 +/- 4 yr, BMI 24 +/- 2 kg/m2, Hb A1C 6.7 +/- 0.1%), and 11 CC patients (age 35 +/- 3 yr, BMI 25 +/- 1 kg/m2, Hb A1C 9.6 +/- 0.1%) underwent two separate randomized, single-blind, 2-h hyperinsulinemic euglycemic clamp studies with (EPI) and without (NO EPI) epinephrine infusion. Epinephrine levels during EPI were similar in all groups (5,197 +/- 344 pmol/l). Glucose (5.3 +/- 0.1 mmol/l) and insulin levels (515 +/- 44 pmol/l) were similar in all groups during the glucose clamps. Endogenous glucose production (EGP) and glucose uptake (R(d)) were determined using [3-H3]glucose. Muscle biopsy was performed at the end of each study. IC had a significantly reduced EGP and R(d) responses to EPI compared with CC. Glucagon responses to EPI were similarly blunted in both IC and CC. Free fatty acid and glycerol response to EPI was greater in CC compared with IC. There was a significantly greater systolic blood pressure response to EPI in CC. We conclude that, despite similar epinephrine, insulin, and glucose levels, intensively treated T1DM patients had reduced cardiovascular, skeletal muscle, hepatic, and adipose target organ responses to EPI compared with conventionally treated T1DM patients. PMID- 15784646 TI - Inhibition of calpain blocks pancreatic beta-cell spreading and insulin secretion. AB - In addition to promoting insulin secretion, an increase in cytosolic Ca(2+) triggered by glucose has been shown to be crucial for spreading of beta-cells attached on extracellular matrix (804G matrix). Calpains are Ca(2+)-dependent cysteine proteases involved in an extended spectrum of cellular responses, including cytoskeletal rearrangements and vesicular trafficking. The present work aimed to assess whether calpain is also implicated in the process of Ca(2+) induced insulin secretion and spreading of rat pancreatic beta-cells. The results indicate calpain dependency of beta-cell spreading on 804G matrix. Indeed, treatment with three distinct calpain inhibitors (N-Ac-Leu-Leu-norleucinal, calpeptin, and ethyl(+)-(2S,3S)-3-[(S)-3-methyl-1-(3-methylbutylcarbamoyl)butyl carbamoyl]-2-ox-iranecarboxylate) inhibited cell spreading induced by glucose and KCl, whereas cell attachment was not significantly modified. Calpain inhibitors also suppressed glucose- and KCl-stimulated insulin secretion without affecting insulin synthesis. Washing the inhibitor out of the cell culture restored spreading on 804G matrix and insulin secretory response after 24 h. In addition, incubation with calpeptin did not affect insulin secretory response to mastoparan that acts on exocytosis downstream of intracellular calcium [Ca(2+)]i. Finally, calpeptin was shown to affect the [Ca(2+)]i response to glucose but not to KCl. In summary, the results show that inhibition of calpain blocks spreading and insulin secretion of primary pancreatic beta-cells. It is therefore suggested that calpain could be a mediator of Ca(2+)-induced-insulin secretion and beta cell spreading. PMID- 15784647 TI - Morale in the oldest old: the Umea 85+ study. AB - OBJECTIVE: to describe morale among the oldest old, and to investigate which social, functional and medical factors are associated with morale in this population. DESIGN: a cross-sectional study. SETTING: a population-based study in the municipality of Umea, a city in Northern Sweden. SUBJECTS: half of the 85 year-old population, and the total population of 90-year-olds and > or = 95-year olds (95-103) were asked to participate (n = 319) and 238 were interviewed. METHODS: structured interviews and assessments during home visits, interviews with relatives and caregivers and review of medical charts. The 17-item Philadelphia Geriatric Center Morale Scale (PGCMS) was used to measure morale. Participants were assessed with the Barthel Activities of Daily Living (ADL) Index, Geriatric Depression Scale (GDS-15), Mini-Mental State Examination (MMSE), Mini Nutritional Assessment (MNA), and a symptom questionnaire. Multiple regression analyses were conducted to find independent factors to explain the variation in the PGCMS score. RESULTS: eighty-four per cent (n = 199) of those interviewed answered the PGCMS. Three-quarters had middle range or high morale. GDS score, type of housing, previous stroke, loneliness and number of symptoms, adjusted for age group and sex, explained 49.3% of the variance of total PGCMS score. CONCLUSIONS: a large proportion of the oldest old had high morale. The most important factors for high morale were the absence of depressive symptoms, living in ordinary housing, having previously had a stroke and yet still living in ordinary housing, not feeling lonely and low number of symptoms. The PGCMS seems applicable in the evaluation of morale among the oldest old. PMID- 15784648 TI - Cyclooxygenase-2 inhibitor SC-236 [4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1 pyrazol-1-l] benzenesulfonamide] suppresses nuclear factor-kappaB activation and phosphorylation of p38 mitogen-activated protein kinase, extracellular signal regulated kinase, and c-Jun N-terminal kinase in human mast cell line cells. AB - SC-236 [4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1-pyrazol-1-l] benzenesulfonamide; C16H11ClF3N3O2S] is a highly selective cyclooxygenase (COX)-2 inhibitor. However, the exact mechanism that accounts for the anti-inflammatory effect of SC-236 is not completely understood. The aim of the present study was to elucidate whether and how SC-236 modulates the inflammatory reaction in a stimulated human mast cell (HMC) line, HMC-1. SC-236 inhibited the expression of tumor necrosis factor-alpha, interleukin (IL)-6, IL-8, vascular endothelial growth factor, COX-2, inducible nitric-oxide synthase, and hypoxia-inducible factor-1alpha in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI)-stimulated HMC-1. SC-236 suppressed nuclear factor (NF)-kappaB activation induced by PMACI, leading to suppression of IkappaB-alpha phosphorylation and degradation. SC-236 also suppressed strong induction of NF-kappaB promoter mediated luciferase activity. In addition, SC-236 suppressed PMACI-induced phosphorylation of the mitogen-activated protein kinase p38, the extracellular regulated kinase p44, and the c-Jun N-terminal kinase and induced expression of mitogen-activated protein kinase phosphatase-1. These results provide new insight into the pharmacological actions of SC-236 as a potential molecule for therapy of mast cell-mediated inflammatory diseases. PMID- 15784649 TI - Prolonged positive modulation of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptors induces calpain-mediated PSD-95/Dlg/ZO-1 protein degradation and AMPA receptor down-regulation in cultured hippocampal slices. AB - Prolonged exposure of cultured hippocampal slices to CX614 [2H,3H,6aH pyrrolidino[2'',1''-3',2']1,3-oxazino[6',5'-5,4]-benzo[e]1,4-dioxan 10-one], a positive alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor (AMPAr) modulator, decreases receptor response to synaptic stimulation, an effect that could reflect reduced receptor expression. The present study investigates this down-regulation and its underlying mechanisms using cultured rat hippocampal slices. Chronic treatment with CX614 gradually reduced levels of glutamate receptor (GluR)1 and GluR2/3 AMPAr subunits and of their anchoring proteins synapse-associated protein 97 (SAP97) and glutamate receptor interacting protein 1 (GRIP1) through 48 h. Decline in SAP97 and GRIP1 levels was associated with increased abundance of lower molecular weight bands, suggesting degradation of these proteins. CX614 effects were partially reversible after drug removal. GluR1 and GluR2/3 down-regulation and their slow recovery were associated with similar changes in SAP97 and GRIP1 levels. Treatment with CX614 for 48 h significantly reduced AMPAr mRNA levels in hippocampus, whereas 8-h exposure did not. Blockade of ionotropic glutamate receptors prevented CX614-induced decrease in AMPAr subunits and mRNA, with regional selectivity, although an AMPAr blocker was more efficacious than an N-methyl-D-aspartate receptor blocker. Blockade of calpain activity reduced CX614-induced degradation of SAP97 and GRIP1 and prevented decreases in AMPAr subunit but not mRNA levels. Treatment with CX614 alone or in combination with glutamate receptor blockers or calpain inhibitor III did not modify lactate dehydrogenase release into culture medium, implying the absence of cell toxicity. We conclude that CX614-induced AMPAr protein loss is primarily mediated by AMPAr activation and involves calpain-dependent proteolysis of SAP97 and GRIP1. CX614-induced suppression of AMPAr gene expression is, however, calpain-independent, and all these effects are not associated with cell damage. PMID- 15784650 TI - CYP3A4 substrate selection and substitution in the prediction of potential drug drug interactions. AB - The complexity of in vitro kinetic phenomena observed for CYP3A4 substrates (homo or heterotropic cooperativity) confounds the prediction of drug-drug interactions, and an evaluation of alternative and/or pragmatic approaches and substrates is needed. The current study focused on the utility of the three most commonly used CYP3A4 in vitro probes for the prediction of 26 reported in vivo interactions with azole inhibitors (increase in area under the curve ranged from 1.2 to 24, 50% in the range of potent inhibition). In addition to midazolam, testosterone, and nifedipine, quinidine was explored as a more "pragmatic" substrate due to its kinetic properties and specificity toward CYP3A4 in comparison with CYP3A5. Ki estimates obtained in human liver microsomes under standardized in vitro conditions for each of the four probes were used to determine the validity of substrate substitution in CYP3A4 drug-drug interaction prediction. Detailed inhibitor-related (microsomal binding, depletion over incubation time) and substrate-related factors (cooperativity, contribution of other metabolic pathways, or renal excretion) were incorporated in the assessment of the interaction potential. All four CYP3A4 probes predicted 69 to 81% of the interactions with azoles within 2-fold of the mean in vivo value. Comparison of simple and multisite mechanistic models and interaction prediction accuracy for each of the in vitro probes indicated that midazolam and quinidine in vitro data provided the best assessment of a potential interaction, with the lowest bias and the highest precision of the prediction. Further investigations with a wider range of inhibitors are required to substantiate these findings. PMID- 15784651 TI - Characterization of the formation and localization of sulfamethoxazole and dapsone-associated drug-protein adducts in human epidermal keratinocytes. AB - Sulfonamide- and sulfone-induced hypersensitivity reactions are thought to be mediated through bioactivation of parent drug molecule(s) to their respective reactive metabolite(s). Recent studies have demonstrated that keratinocytes can bioactivate sulfonamides and sulfones. Using enzyme-linked immunosorbent assay and hapten-specific rabbit antisera developed in our laboratory, we found that incubation of either normal human epidermal keratinocytes (NHEKs) or an immortalized human keratinocyte cell line (HaCaT) with sulfamethoxazole (SMX) or dapsone (DDS) resulted in the formation of drug/metabolite protein adducts. The formation of these adducts with SMX was increased in the presence of ascorbic acid, whereas N-acetylcysteine decreased adduct formation with both SMX and DDS. Adduct formation was confirmed using confocal microscopy when NHEKs were incubated with SMX, DDS, or their respective arylhydroxylamine metabolites. Cellular distribution of adducts was compared in permeable versus nonpermeable NHEKs. Exposure to SMX, DDS, or dapsone hydroxylamine resulted in the formation of intracellular adducts, whereas SMX hydroxylamine also resulted in the presence of adducts on the cell surface. In summary, our work shows that keratinocytes can bioactivate SMX/DDS to form drug-protein adducts, which may be acquired by antigen-presenting cells upon keratinocyte cell death, evoking an immune response. In addition, keratinocytes may themselves present antigen to hapten specific cytotoxic T lymphocytes. Furthermore, our results also suggest that different sulfonamides/sulfones may have different protein targets for in situ haptenation in keratinocytes. PMID- 15784652 TI - Acute and chronic corticotropin-releasing factor 1 receptor blockade inhibits cocaine-induced dopamine release: correlation with dopamine neuron activity. AB - Corticotropin-releasing factor (CRF) is a neuropeptide associated with the integration of the physiological and behavioral responses to stress. Recently, CRF1 receptor antagonists have been shown to decrease cocaine self-administration and inhibit stress-induced reinstatement of cocaine-seeking behavior. The exact mechanisms underlying this effect are not clear. Based on the large amount of literature demonstrating an association between dopaminergic neurotransmission and reward-related behavior, the aim of the present study was to examine the effects of acute versus chronic CRF1 receptor blockade on mesencephalic dopamine (DA) neuron activity (determined by in vivo extracellular recordings) and extracellular DA levels in the nucleus accumbens (Acb) (using in vivo microdialysis). In addition, the effect of CRF1 receptor antagonism on cocaine induced DA overflow in the Acb was examined and correlated with DA neuron activity in the ventral tegmental area (VTA). Acute (but not chronic) CRF1 receptor blockade by CRA-0450 [1-[8-(2,4-dichlorophenyl)-2-methylquinolin-4-yl] 1,2,3,6-tetrahydropyridine-4-carboxamide benzenesulfonate] was found to significantly increase DA neuron population activity without affecting burst firing, average firing rate, or Acb DA levels. In addition, both acute and chronic CRF1 receptor antagonism significantly reduced cocaine-stimulated DA overflow in the Acb, and this reduction was correlated with an attenuated cocaine induced inhibition of DA population activity. Taken as a whole, these data demonstrate that, although DA neuron population activity exhibits tolerance to chronic CRF1 receptor antagonism (by CRA-0450), tolerance does not develop to the selective inhibition of cocaine-induced DA release (in the Acb) and, as such, may be beneficial in the treatment of cocaine addiction. PMID- 15784653 TI - Protective mechanisms of a metalloporphyrinic peroxynitrite decomposition catalyst, WW85, in rat cardiac transplants. AB - Nitric oxide (NO) derived from inducible NO synthase has been implicated in cardiac rejection. However, little is known about the role of the reactive nitrogen species peroxynitrite. We examined the protective actions of a peroxynitrite decomposition catalyst, WW85, in an experimental model of acute cardiac rejection. Heterotopic, abdominal transplantation of rat donor hearts was performed. Groups included isografts, allografts, or allografts treated with WW85, cyclosporine, or cyclosporine + WW85. We determined graft survival, histological rejection, and graft function (by in situ sonomicrometry). Intragraft biochemical analysis of cytokines and proapoptotic and antiapoptotic gene expression using reverse transcriptase-polymerase chain reaction were determined. Treatment with WW85 or cyclosporine alone prolonged graft survival, improved graft function, and decreased histological rejection. Graft survival was further significantly (P < 0.001) enhanced by combination treatment. A decrease was also shown in nitrotyrosine, poly(ADP-ribose) polymerase (PARP) activation, and lipid peroxide formation by WW85 that was potentiated when given in combination with cyclosporine. Benefits could not be ascribed to changes in intragraft myeloperoxidase activity. Only combination therapy produced significant decreases in inflammatory cytokine gene expression, suggesting that WW85 acted primarily downstream of these stimuli. In general, WW85 had no direct action on expression of the proapoptotic gene, Fas ligand; however, WW85 given alone or with cyclosporine enhanced expression of antiapoptotic genes Bcl-2 and Bcl-xL. Collectively, these findings suggest a protective action of the peroxynitrite decomposition catalyst WW85 on graft rejection that is independent of any action on leukocyte sequestration and cytokine gene expression. Rather, effects seem to be downstream on decreased protein nitration, decreased lipid peroxidation, and decreased PARP activation. PMID- 15784654 TI - A novel effect for annexin 1-derived peptide ac2-26: reduction of allergic inflammation in the rat. AB - Previous investigations have provided evidence that the N-terminal peptide of annexin 1 (peptide Ac2-26) has the capacity of reproducing the anti-inflammatory actions of the full-length protein in many systems. In the current study, we report the effectiveness of the peptide Ac2-26 as an antiallergic tool in a model of rat pleurisy and provide indication for some of the mechanisms involved. In rats inflamed by injection of ovalbumin into the pleural cavity 14 days postsensitization, peptide Ac2-26 (50-200 microg/cavity) inhibited mast cell degranulation, plasma protein leakage, and the accumulation of both neutrophils and eosinophils. Treatment with either peptide Ac2-26 (200 microg/cavity) or dexamethasone (1 mg/kg i.p.) inhibited ovalbumin-induced eotaxin release in the pleural effluents. In vitro, peptide Ac2-26 inhibited ovalbumin-evoked histamine release from subcutaneous tissue fragments obtained from sensitized rats (33-66 microM) and interleukin-13-evoked eotaxin generation from cultured rat mesothelial cells (16-33 microM) but not eosinophil chemotaxis. This work demonstrates that the annexin 1 mimetic peptide Ac2-26 prevents allergen-evoked eosinophilic inflammatory response in rats. Combined analysis of the in vivo and in vitro experiments presented herein suggests that the blockade of secretion of pivotal mediators for the allergic response, such as histamine and eotaxin, could be responsible for the inhibitory actions displayed by peptide Ac2-26. PMID- 15784655 TI - YC-1 [3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole] inhibits endothelial cell functions induced by angiogenic factors in vitro and angiogenesis in vivo models. AB - Angiogenesis is a process that involves endothelial cell proliferation, migration, invasion, and tube formation, and inhibition of these processes has implications for angiogenesis-mediated disorders. The purpose of this study was to evaluate the antiangiogenic efficacy of YC-1 [3-(5'-hydroxymethyl-2'-furyl)-1 benzyl indazole] in well characterized in vitro and in vivo systems. YC-1 inhibited the ability of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in a dose-dependent manner to induce proliferation, migration, and tube formation in human umbilical vascular endothelial cells; these outcomes were evaluated using [3H]thymidine incorporation, transwell chamber, and Matrigel-coated slide assays, respectively. YC-1 inhibited VEGF- and bFGF-induced p42/p44 mitogen-activated protein kinase and Akt phosphorylation as well as protein kinase C alpha translocation using Western blot analysis. The effect of YC-1 on angiogenesis in vivo was evaluated using the mouse Matrigel implant model. YC-1 administered orally in doses of 1 to 100 mg/kg/day inhibited VEGF- and bFGF-induced neovascularization in a dose dependent manner over 7 days. These results indicate that YC-1 has antiangiogenic activity at very low doses. Moreover, in transplantable murine tumor models, YC-1 administered orally displayed a high degree of antitumor activity (treatment-to control life span ratio > 175%) without cytotoxicity. YC-1 may be useful for treating angiogenesis-dependent human diseases such as cancer. PMID- 15784656 TI - Hepatic glucocorticoid receptor antagonism is sufficient to reduce elevated hepatic glucose output and improve glucose control in animal models of type 2 diabetes. AB - Glucocorticoids amplify endogenous glucose production in type 2 diabetes by increasing hepatic glucose output. Systemic glucocorticoid blockade lowers glucose levels in type 2 diabetes, but with several adverse consequences. It has been proposed, but never demonstrated, that a liver-selective glucocorticoid receptor antagonist (LSGRA) would be sufficient to reduce hepatic glucose output (HGO) and restore glucose control to type 2 diabetic patients with minimal systemic side effects. A-348441 [(3b,5b,7a,12a)-7,12-dihydroxy-3-{2-[{4 [(11b,17b)-17-hydroxy-3-oxo-17-prop-1-ynylestra-4,9-dien-11-yl] phenyl}(methyl)amino]ethoxy}cholan-24-oic acid] represents the first LSGRA with significant antidiabetic activity. A-348441 antagonizes glucocorticoid-up regulated hepatic genes, normalizes postprandial glucose in diabetic mice, and demonstrates synergistic effects on blood glucose in these animals when coadministered with an insulin sensitizer. In insulin-resistant Zucker fa/fa rats and fasted conscious normal dogs, A-348441 reduces HGO with no acute effect on peripheral glucose uptake. A-348441 has no effect on the hypothalamic pituitary adrenal axis or on other measured glucocorticoid-induced extrahepatic responses. Overall, A-348441 demonstrates that an LSGRA is sufficient to reduce elevated HGO and normalize blood glucose and may provide a new therapeutic approach for the treatment of type 2 diabetes. PMID- 15784657 TI - "Culture of life" politics at the bedside--the case of Terri Schiavo. PMID- 15784658 TI - Terri Schiavo--a tragedy compounded. PMID- 15784659 TI - Images in clinical medicine. Streptococcal pharyngitis. PMID- 15784660 TI - What ails the FDA? PMID- 15784661 TI - 2015--the future of medical libraries. PMID- 15784662 TI - Quiet in the library. PMID- 15784663 TI - Transmission of rabies virus from an organ donor to four transplant recipients. AB - BACKGROUND: In 2004, four recipients of kidneys, a liver, and an arterial segment from a common organ donor died of encephalitis of an unknown cause. METHODS: We reviewed the medical records of the organ donor and the recipients. Blood, cerebrospinal fluid, and tissues from the recipients were tested with a variety of assays and pathological stains for numerous causes of encephalitis. Samples from the recipients were also inoculated into mice. RESULTS: The organ donor had been healthy before having a subarachnoid hemorrhage that led to his death. Encephalitis developed in all four recipients within 30 days after transplantation and was accompanied by rapid neurologic deterioration characterized by agitated delirium, seizures, respiratory failure, and coma. They died an average of 13 days after the onset of neurologic symptoms. Mice inoculated with samples from the affected patients became ill seven to eight days later, and electron microscopy of central nervous system (CNS) tissue demonstrated rhabdovirus particles. Rabies-specific immunohistochemical and direct fluorescence antibody staining demonstrated rabies virus in multiple tissues from all recipients. Cytoplasmic inclusions consistent with Negri bodies were seen in CNS tissue from all recipients. Antibodies against rabies virus were present in three of the four recipients and the donor. The donor had told others of being bitten by a bat. CONCLUSIONS: This report documenting the transmission of rabies virus from an organ donor to multiple recipients underscores the challenges of preventing and detecting transmission of unusual pathogens through transplantation. PMID- 15784664 TI - The serotonin syndrome. PMID- 15784665 TI - Images in clinical medicine. Acquired acrodermatitis enteropathica. PMID- 15784666 TI - Case records of the Massachusetts General Hospital. Case 8-2005. A 10-year-old boy with pain in the right thigh. PMID- 15784667 TI - Deadweight?--The influence of obesity on longevity. PMID- 15784668 TI - A potential decline in life expectancy in the United States in the 21st century. AB - Forecasts of life expectancy are an important component of public policy that influence age-based entitlement programs such as Social Security and Medicare. Although the Social Security Administration recently raised its estimates of how long Americans are going to live in the 21st century, current trends in obesity in the United States suggest that these estimates may not be accurate. From our analysis of the effect of obesity on longevity, we conclude that the steady rise in life expectancy during the past two centuries may soon come to an end. PMID- 15784669 TI - Anemia and gene therapy--a matter of control. PMID- 15784670 TI - Ablation for atrial fibrillation in congestive heart failure. PMID- 15784671 TI - The "pill-in-the-pocket" approach to atrial fibrillation. PMID- 15784672 TI - Vascular events after acute infection or vaccination. PMID- 15784673 TI - Altered nuclear transfer. PMID- 15784674 TI - Acute pericarditis. PMID- 15784676 TI - H1-antihistamines. PMID- 15784675 TI - Medical discoveries and scientific priority. PMID- 15784677 TI - Leflunomide for polyomavirus type BK nephropathy. PMID- 15784678 TI - NACA is a positive regulator of human erythroid-cell differentiation. AB - We have previously identified the transcript encoding NACA (the alpha chain of the nascent-polypeptide-associated complex) as a cytokine-modulated specific transcript in the human TF-1 erythroleukemic cell line. This protein was already known to be a transcriptional co-activator that acts by potentiating AP-1 activity in osteoblasts, and is known to be involved in the targeting of nascent polypeptides. In this study, we investigate the role of NACA in human hematopoiesis. Protein distribution analyses indicate that NACA is expressed in undifferentiated TF-1 cells and in human-cord-blood-derived CD34(+) progenitor cells. Its expression is maintained during in vitro erythroid differentiation but, in marked contrast, its expression is suppressed during their megakaryocytic or granulocytic differentiation. Ectopic expression of NACA in CD34(+) cells under culture conditions that induce erythroid-lineage differentiation leads to a marked acceleration of erythroid-cell differentiation. Moreover, ectopic expression of NACA induces erythropoietin-independent differentiation of TF-1 cells, whereas downregulation of NACA by RNA interference abolishes the induction of hemoglobin production in these cells and diminishes glycophorin-A (GPA) expression by CD34(+) progenitors cultured under erythroid differentiation conditions. Altogether, these results characterize NACA as a new factor involved in the positive regulation of human erythroid-cell differentiation. PMID- 15784679 TI - STAT-1 facilitates the ATM activated checkpoint pathway following DNA damage. AB - STAT-1 plays a role in mediating stress responses to various stimuli and has also been implied to be a tumour suppressor. Here, we report that STAT-1-deficient cells have defects both in intra-S-phase and G2-M checkpoints in response to DNA damage. Interestingly, STAT-1-deficient cells showed reduced Chk2 phosphorylation on threonine 68 (Chk2(-T68)) following DNA damage, suggesting that STAT-1 might function in the ATM-Chk2 pathway. Moreover, the defects in Chk2(-T68) phosphorylation in STAT-1-deficient cells also correlated with reduced degradation of Cdc25A compared with STAT-1-expressing cells after DNA damage. We also show that STAT-1 is required for ATM-dependent phosphorylation of NBS1 and p53 but not for BRCA1 or H2AX phosphorylation following DNA damage. Expression levels of BRCT mediator/adaptor proteins MDC1 and 53BP1, which are required for ATM-mediated pathways, are reduced in cells lacking STAT-1. Enforced expression of MDC1 into STAT-1-deficient cells restored ATM-mediated phosphorylation of downstream substrates. These results imply that STAT-1 plays a crucial role in the DNA-damage-response by regulating the expression of 53BP1 and MDC1, factors known to be important for mediating ATM-dependent checkpoint pathways. PMID- 15784680 TI - Microtubule nucleation and anchoring at the centrosome are independent processes linked by ninein function. AB - The centrosome organizes microtubules by controlling nucleation and anchoring processes. In mammalian cells, subdistal appendages of the mother centriole are major microtubule-anchoring structures of the centrosome. It is not known how newly nucleated microtubules are anchored to these appendages. We show here that ninein, a component of subdistal appendages, localizes to the centriole via its C terminus and interacts with gamma-tubulin-containing complexes via its N terminus. Expression of a construct encoding the ninein C-terminus displaced endogenous ninein and the gamma-tubulin ring complex (gamma-TuRC) from the centrosome, leading to microtubule nucleation and anchoring defects. By contrast, expression of a fusion consisting of the N- and C-terminal domains (lacking the central coiled-coil region) displaced endogenous ninein without perturbing gamma TuRC localization. Accordingly, only anchoring defects were observed in this case. Therefore, expression of this fusion appeared to uncouple microtubule nucleation and anchorage activities at the centrosome. Our results suggest that ninein has a role not only in microtubule anchoring but also in promoting microtubule nucleation by docking the gamma-TuRC at the centrosome. In addition, we show that the gamma-TuRC might not be sufficient to anchor microtubules at the centrosome in the absence of ninein. We therefore propose that ninein constitutes a molecular link between microtubule-nucleation and -anchoring activities at the centrosome. PMID- 15784681 TI - Protein synthesis inhibitors and the chemical chaperone TMAO reverse endoplasmic reticulum perturbation induced by overexpression of the iodide transporter pendrin. AB - An outcome of overloading of the endoplasmic reticulum (ER) folding machinery is a perturbation in ER function and the formation of intracellular aggregates. The latter is a key pathogenic factor in numerous diseases known as ER storage diseases. Here, we report that heterologous overexpression of the green fluorescent protein-tagged iodide transporter pendrin (GFP-PDS) perturbs folding and degradation processes in the ER. Pendrin (PDS) is a chloride-iodide transporter found in thyroid cells. Mutations in PDS can cause its retention in the ER and are associated with Pendred syndrome. Biochemical and live-cell analyses demonstrated that wild-type GFP-PDS is predominantly retained in perinuclear aggregates and in ER membranes, causing their collapse and vesiculation. Inhibition of protein synthesis by cycloheximide (CHX) or puromycin caused dissociation of the GFP-PDS aggregates and returned the ER to its normal reticular morphology. Blocking protein synthesis promoted folding and export of ER-retained GFP-PDS, as demonstrated by surface-biotinylation analysis and by CHX or puromycin-induced accumulation of YFP-PDS in the Golgi apparatus during a 20 degrees C temperature-block experiment. The chemical chaperone trimethylamine-N oxide (TMAO) also reversed the GFP-PDS-mediated ER collapse and vesiculation, suggesting that exposed hydrophobic stretches of misfolded or aggregated GFP-PDS may contribute to ER retention. These data suggest that GFP-PDS is a slow-folding protein with a propensity to form aggregates when overexpressed. Thus, we describe a system for the reversible induction of ER stress that is based entirely on the heterologous overexpression of GFP-PDS. PMID- 15784682 TI - Detection of HSP60 on the membrane surface of stressed human endothelial cells by atomic force and confocal microscopy. AB - The highly conserved and ubiquitous heat shock proteins (HSP) are essential for the cellular homeostasis and efficiently trigger cellular responses to stress conditions. Both microbial and human HSP act as dominant antigens in numerous infectious and autoimmune diseases such as atherosclerosis, inducing a strong immune-inflammatory response. In the present study, the surface localization of HSP60 on stressed and unstressed human umbilical venous endothelial cells (HUVECs) was investigated using sensitive high resolution microscopy methods and flow cytometry. Confocal laser scanning microscopy (CLSM) revealed an increase of HSP60 in the mitochondria and on the surface of heat-stressed living and fixed HUVECs compared to unstressed cells. Atomic force microscopy (AFM), which has developed as sensitive surface-probe technique in biology, confirmed the presence of HSP60 on the membrane of stressed cells at an even higher lateral resolution by detecting specific single molecule binding events between the monoclonal antibody AbII-13 tethered to AFM tips and HSP60 molecules on cells. The interaction force (force required to break a single AbII-13/HSP60 bond) was 59+/ 2 pN, which correlated nicely to the 51+/-1 pN measured with isolated HSP60 attached to mica surfaces. Overall, we found clear evidence for the occurrence of HSP60 on the surface of stressed HUVECs in a very similar patchy distribution pattern in living and fixed cells. The relevance of our findings with respect to the role of HSP60 in atherogenesis is discussed. PMID- 15784683 TI - DNA CpG hypomethylation induces heterochromatin reorganization involving the histone variant macroH2A. AB - In mammalian heterochromatin, cytosine bases of CpG dinucleotides are symmetrically modified by methylation. Patterns of CpG methylation are maintained by the action of Dnmt1, the mammalian maintenance cytosine methyltransferase enzyme. We genetically manipulated the levels of CpG methylation and found that extensive chromatin alterations occur in pericentric heterochromatin. Homozygous mutations in Dnmt1 cause severe hypomethylation of pericentric heterochromatin and concomitant chromatin reorganization involving the histone variant macroH2A. Demethylation-induced alterations in macroH2A localization occur in both interphase and mitotic embryonic stem (ES) cells. Heterochromatin protein 1 (HP1) marks interphase pericentric heterochromatin (chromocenters). MacroH2A immunostaining in Dnmt1(-/-) cells becomes coincident with chromocenters detected by HP1 content. MacroH2A, but not HP1, is enriched in nuclease-resistant chromatin fractions extracted from Dnmt1(-/-) cells. Normal localization of macroH2A was restored upon reintroduction of a Dnmt1 transgene into Dnmt1(-/-) cells. MacroH2A localization was also affected in T-antigen-transformed fibroblasts subjected to the conditional mutation of Dnmt1. Together, these results suggest that pericentric heterochromatin can be maintained in the absence of CpG methylation, but in a significantly altered configuration. PMID- 15784684 TI - Interplay between septin organization, cell cycle and cell shape in yeast. AB - Septins are conserved filament-forming proteins that assemble into cortical cytoskeletal structures in animal and fungal cells. Although rapid progress has been made into the functions of septins, the mechanisms governing their localization and organization remain mysterious. In Saccharomyces cerevisiae, Cdc42p organizes the septin cytoskeleton into a ring in preparation for bud formation, following which septins remain as a collar at the mother-bud neck. We have dissected the phenotype of cdc42(V36T,K94E) cells that display an aberrant cell shape correlated with the development of ectopic septin caps and rings within the bud. The results suggest that a well-assembled septin cortex plays a novel role in directing growth to shape the nascent bud, and that a disorganized septin cortex directs improper growth generating an aberrant neck. Conversely, we found that the elongated bud shape arising as a result of the morphogenesis checkpoint cell cycle delay that accompanies septin perturbation can feed back to exacerbate minor defects in septin organization, by maintaining a bud-tip localized septin assembly activity that competes with the neck-localized septin cortex. Using this exacerbation as a tool, we uncovered septin organization defects in many mutants not previously known to display such defects, expanding the cast of characters involved in proper assembly of the septin cortex to include CLN1, CLN2, BNI1, BNI4, BUD3, BUD4 and BUD5. PMID- 15784685 TI - Classical protein kinase C(s) regulates targeting of synaptotagmin IX to the endocytic recycling compartment. AB - Neuronal and non-neuronal tissues show distinctly different intracellular localization of synaptotagmin (Syt) homologues. Therefore, cell type-specific mechanisms are likely to direct Syt homologues to their final cellular destinations. Syt IX localizes to dense core vesicles in PC12 cells. However, in the rat basophilic leukemia (RBL-2H3) mast cell line, as well as in CHO cells, Syt IX is localized at the endocytic recycling compartment (ERC). We show that targeting of Syt IX to the ERC involves constitutive trafficking to the plasma membrane followed by internalization and transport to the ERC. We further show that internalization from the plasma membrane and delivery to the ERC are dependent on phosphorylation by Ca(2+)-dependent protein kinase Calpha or beta. As such, correct targeting of Syt IX is facilitated by the phorbol ester TPA but prevented by the cPKC inhibitor Go 6976. PMID- 15784689 TI - Inhibition of IKK down-regulates antigen + IgE-induced TNF production by mast cells: a role for the IKK-IkappaB-NF-kappaB pathway in IgE-dependent mast cell activation. AB - Mast cells (MC) are major effector cells for allergic diseases. Cross-linking of immunoglobulin E (IgE) and its high-affinity receptor, FcepsilonRI, by antigen initiates a cascade of signaling events leading to nuclear factor (NF)-kappaB activation and tumor necrosis factor (TNF) production. Here, we demonstrated that inhibition of inhibitor of kappaB (IkappaB) kinase (IKK) by a peptide IKK inhibitor or by four individual chemical IKK inhibitors including 15-deoxy prostaglandin J(2), BMS-345541, SC-514, or sulindac significantly blocked IgE + trinitrophenyl (TNP)-induced TNF production by mouse bone marrow-derived MC (BMMC). Moreover, IgE + TNP induced a rapid phosphorylation of IKKalpha but not IKKbeta in BMMC. IgE + TNP-induced phosphorylation of IKKalpha was accompanied with phosphorylation and degradation of IkappaBalpha, subsequent NF-kappaB activation, and TNF production. Inhibition of IKK by sulindac decreased IKKalpha phosphorylation, IkappaBalpha phosphorylation and degradation, NF-kappaB activation, and TNF production by BMMC. It is interesting that IgE + TNP stimulation also induced a prominent synthesis of IKKalpha and IkappaBalpha. Inhibition of NF-kappaB activity by pyrrolidine dithiocarbomate (PDTC) blocked IgE + TNP-induced IkappaBalpha synthesis. NF-kappaB activity and TNF production were also inhibited when PDTC was used even after IgE + TNP stimulation, suggesting a potential role for the newly synthesized IkappaBalpha in MC activation. In addition, IgE + TNP-induced IKKalpha and IkappaBalpha phosphorylation was inhibited by a protein kinase C (PKC) inhibitor Ro 31-8220. Taken together, our results support a role for the IKK-IkappaB-NF-kappaB pathway, which likely involves PKC in IgE-dependent TNF production by MC. Thus, IKK may serve as a new target for the regulation of MC function in allergy. PMID- 15784688 TI - Evaluation of normal and neoplastic human mast cells for expression of CD172a (SIRPalpha), CD47, and SHP-1. AB - Signal regulatory proteins (SIRPs) and tyrosine phosphatases have recently been implicated in the control of receptor tyrosine kinase (RTK)-dependent cell growth. In systemic mastocytosis (SM), neoplastic cells are driven by the RTK KIT, which is mutated at codon 816 in most patients. We examined expression of SIRPalpha, SIRPalpha ligand CD47, and Src homology 2 domain-containing protein tyrosine phosphatase-1 (SHP-1), a tyrosine phosphatase-type, negative regulator of KIT-dependent signaling, in normal human lung mast cells (HLMC) and neoplastic MC obtained from nine patients with SM. As assessed by multicolor flow cytometry, normal LMC expressed SIRPalpha, CD47, and SHP-1. In patients with SM, MC also reacted with antibodies against SIRPalpha and CD47. By contrast, the levels of SHP-1 were low or undetectable in MC in most cases. Corresponding data were obtained from mRNA analysis. In fact, whereas SIRPalpha mRNA and CD47 mRNA were detected in all samples, the levels of SHP-1 mRNA varied among donors. To demonstrate adhesive functions for SIRPalpha and CD47 on neoplastic MC, an adhesion assay was applied using the MC leukemia cell line HMC-1, which was found to bind to immobilized extracellular domains of SIRPalpha1 (SIRPalpha1ex) and CD47 (CD47ex), and binding of these cells to CD47ex was inhibited by the CD172 antibody SE5A5. In summary, our data show that MC express functional SIRPalpha and CD47 in SM, whereas expression of SHP-1 varies among donors and is low compared with LMC. It is hypothesized that CD172 and CD47 contribute to MC clustering and that the "lack" of SHP-1 in MC may facilitate KIT-dependent signaling in a subgroup of patients. PMID- 15784687 TI - Involvement of CC chemokine ligand 18 (CCL18) in normal and pathological processes. AB - CC chemokine ligand 18 (CCL18) was originally discovered as pulmonary and activation-regulated chemokine (PARC), dendritic cell (DC)-chemokine 1 (DC-CK1), alternative macrophage activation-associated CC chemokine-1 (AMAC-1), and macrophage inflammatory protein-4 (MIP-4). CCL18 primarily targets lymphocytes and immature DC, although its agonistic receptor remains unknown so far. CCL18 is mainly expressed by a broad range of monocytes/macrophages and DC. A more profound understanding of the various activation programs and functional phenotypes of these producer cells might give a better insight in the proinflammatory versus anti-inflammatory role of this CC chemokine. It is interesting that CCL18 is constitutively present at high levels in human plasma and likely contributes to the physiological homing of lymphocytes and DC and to the generation of primary immune responses. Furthermore, enhanced CCL18 production has been demonstrated in several diseases, including various malignancies and inflammatory joint, lung, and skin diseases. The lack of a rodent counterpart for human CCL18 sets all hope on primate animal models to further elucidate the importance of CCL18 in vivo. This review will address these different aspects in more detail. PMID- 15784691 TI - Radiosensitivity of lymphoblastoid cell lines with a heterozygous BRCA1 mutation is not detected by the comet assay and pulsed field gel electrophoresis. AB - Lymphoblastoid cell lines (LCL) with a heterozygous mutation in the breast cancer susceptibility gene BRCA1 have been repeatedly used to elucidate the biological consequences of such a mutation with respect to radiation sensitivity and DNA repair deficiency. Our previous results indicated that LCL with a BRCA1 mutation do not generally show the same chromosomal mutagen sensitivity in the micronucleus test as lymphocytes with the same BRCA1 mutation. To further study the radiosensitivity of LCL with a BRCA1 mutation, we now performed comparative investigations with the alkaline (pH 13) and the neutral (pH 8.3) comet assay and pulsed field gel electrophoresis (PFGE). These tests are commonly used to determine the repair capacity for DNA double strand breaks (DNA-DSB). Six LCL (three established from women with a heterozygous BRCA1 mutation and three from healthy controls) were investigated. Induction (2 and 5 Gy) of gamma-ray-induced DNA damage and its repair (during 60 min after irradiation) was measured with the alkaline and neutral comet assay. Comparative experiments were performed with PFGE determining the induction of DNA-DSB by 10-50 Gy gamma-irradiation and their repair during 6 h. There was no significant difference between LCL with and without BRCA1 mutation in any of these experiments. Therefore, using these methods, no indication for a delayed repair of DNA-DSB in LCL with a BRCA1 mutation was found. However, these results do not generally exclude DNA-DSB repair deficiency in these cell lines because the methods applied have limited sensitivity and only measure the speed but not the fidelity of the repair process. PMID- 15784690 TI - Zidovudine induces S-phase arrest and cell cycle gene expression changes in human cells. AB - Antiretroviral therapy for the human immunodeficiency virus-1 (HIV-1) typically includes two nucleoside reverse transcriptase inhibitors (NRTIs). 3'-Azido-3' deoxythymidine (AZT, Zidovudine) plus 2'-deoxy-3'-thiacytidine (3TC, Lamivudine) is a combination that is used frequently. The NRTIs are mutagenic nucleoside analogs that become incorporated into DNA and terminate replication. We therefore hypothesized that exposure to this class of drug may alter cell cycle parameters. We used flow cytometry to examine the cell cycle in human epithelioid carcinoma (HeLa) cells exposed to AZT and 3TC alone, as well as a series of AZT/3TC dose combinations: (A) 125.0 microM AZT/12.5 microM 3TC; (B) 250.0 microM AZT/25.0 microM 3TC; and (C) 500 microM AZT/50 microM 3TC. At 24 h, at all doses, there was a good cell viability (>/=68%), and incorporation of AZT into nuclear DNA. Using flow cytometry, a dose-related increase in the percentage of cells in S phase, from 9.5% with no drug, to 36.0% with dose C, was observed in cells exposed for 24 h (P = 0.001, ANOVA). A concomitant decrease in the percentage of cells in G(1) phase, from 82.6% with no drug to 58.5% with dose C, was observed in cells exposed for 24 h (P = 0.017, ANOVA). A similar S phase arrest was seen in cells exposed to 125, 250 and 500 microM AZT alone, but there was no S phase alteration with 50 microM 3TC alone, suggesting that AZT is responsible for the accumulation of cells in S phase. To elucidate the accumulation of cells in S phase and explore the cell cycle gene expression changes induced by AZT and 3TC, we used c-DNA microarray, Cell Cycle Super Array and real-time PCR. There was a strong upregulation of the DNA damage-inducible transcript 3 (DDIT3 or GADD153) in NRTI-exposed cells. In addition, AZT induced an upregulation of cyclin D1 accompanied by a downregulation of the cyclin D1-associated inhibitors P18 and P57, and the G(1)-S check point gene P21, the net effect of which would be to foster a cell progression into S phase. Cyclin A2 was down-regulated in cells exposed to AZT, suggesting a block in S-G(2)-M progression that would also be consistent with the accumulation of cells in S phase. Overall, the study demonstrates that AZT, but not 3TC, causes an arrest of cells in S phase with a consistent alteration in the expression of several cell cycle genes. PMID- 15784692 TI - The emperor wears no clothes in the field of carcinogen risk assessment: ignored concepts in cancer risk assessment. AB - The following is a position paper challenging the paradigm that 'carcinogen = mutagen', and that the current rodent bioassay to predict risks to human cancers is relevant and useful. Specifically, we review current observations concerning carcinogenesis that might lead to another approach for assessing the identification of human carcinogenic hazards and the risk assessment that chemicals might pose. We give a brief review of the multistage and multimechanism process of cancer in a tissue that involves not only genotoxic but also epigenetic events, and the importance of stem and progenitor cells in the development of cancer. We focus on the often ignored 'epigenetic' effects of carcinogens and the role of cell communication systems in epigenetically altering gene expression that leads to an imbalance of cell proliferation, differentiation and apoptosis in a tissue that can contribute to the cancer process. To draw attention to the fact that the current paradigm and policy to test toxic chemicals is often misleading and incorrect, we discuss how oxidative stress, in spite of the DNA damaging data, most probably contributes to cancer at the epigenetic level. Additionally, we briefly review how this mutagenic concept has greatly diverted attention away from doing research on the lower molecular weight, non-genotoxic, polycyclic aromatic hydrocarbons (PAHs), and how these low molecular weight PAHs are etiologically more relevant to the disease potential of environmental mixtures such as cigarette smoke. PMID- 15784693 TI - Global gene annotation analysis and transcriptional profiling identify key biological modules in hypoxic pulmonary hypertension. AB - Chronic hypoxic pulmonary hypertension is an important clinical disorder causing significant morbidity. Despite recent discoveries, many molecular mechanisms involved in its pathogenesis remain unexplored. We have undertaken a systematic and unbiased approach to gain global insights into this complex process. By combining transcriptional profiling with rigorous statistical methods and cluster analysis, we identified the dominant temporal patterns of gene expression during progression and regression of hypoxic pulmonary hypertension. We next integrated these results with global gene annotation analysis to identify key biological themes involved in the development and resolution of hypoxic pulmonary hypertension and vascular remodeling. This novel approach assigned biological roles to thousands of candidate genes based on their temporal expression profiles and membership in specific biological modules. Our procedure confirmed several molecular pathways and gene products known to be important in hypoxic pulmonary hypertension. Furthermore, we discovered several novel candidates and molecular mechanisms, including IQ motif containing GTPase-activating protein-1 (IQGAP1), decorin, insulin-like growth factor binding protein-3 (IGFBP3), and lactotransferrin, that may play crucial roles in hypoxic pulmonary hypertension and vascular remodeling. Our methodology of integrating transcriptional profiling, cluster analysis, and global gene annotation provides new insights into the pathophysiology of pulmonary hypertension and is applicable to other models of human disease. PMID- 15784694 TI - Identification of human exons overexpressed in tumors through the use of genome and expressed sequence data. AB - Alternative splicing is one of the major sources of the large transcriptional diversity found in human cells. Splicing variants have been shown to be associated with features like spreading and progression in several human tumors. Therefore, such variants may be of great importance as both diagnostic and therapeutic tools. Here, by using a set of criteria regarding the expression pattern of splicing variants and statistical analyses, we were able to screen the genome for exons overexpressed in tumors of specific tissues. However, as in other analyses attempting to identify tumor-associated variants, our list of candidates was seriously inflated with cases of genes differentially expressed in tumors. To exclude these cases and increase the probability of finding bona fide regulated splicing variants, we performed a serial analysis of gene expression (SAGE), excluding those genes that were shown to be upregulated in tumors. This allowed us to predict the overexpression of single exons in specific tumors. Our final group of candidates includes 1,386 exons belonging to 638 genes. Experimental validation of a few candidates in normal tissue, tumor cell lines, and patient samples suggests that most of these candidates are indeed tumor associated exons. Further functional classification of our candidate genes shows that our final list is slightly inflated with cancer-related genes. PMID- 15784686 TI - Proposed criteria for assessing the efficacy of cancer reduction by plant foods enriched in carotenoids, glucosinolates, polyphenols and selenocompounds. AB - BACKGROUND AND AIMS: The cancer-protective properties of vegetable consumption are most likely mediated through 'bioactive compounds' that induce a variety of physiologic functions including acting as direct or indirect antioxidants, regulating enzymes and controlling apoptosis and the cell cycle. The 'functional food' industry has produced and marketed foods enriched with bioactive compounds, but there are no universally accepted criteria for judging efficacy of the compounds or enriched foods. SCOPE: Carotenoids, glucosinolates, polyphenols and selenocompounds are families of bioactive compounds common to vegetables. Although numerous studies have investigated the agricultural and human health implications of enriching foods with one or more of these compounds, inadequate chemical identification of compounds, lack of relevant endpoints and inconsistencies in mechanistic hypotheses and experimental methodologies leave many critical gaps in our understanding of the benefits of such compounds. This review proposes a decision-making process for determining whether there is reasonable evidence of efficacy for the both the compound and the enriched food. These criteria have been used to judge the evidence of efficacy for cancer prevention by carotenoids, polyphenols, glucosinolates and selenocompounds. CONCLUSIONS: The evidence of efficacy is weak for carotenoids and polyphenols; the evidence is stronger for glucosinolates and lycopene, but production of enriched foods still is premature. Additionally there is unacceptable variability in the amount and chemical form of these compounds in plants. The evidence of efficacy for selenocompounds is strong, but the clinical study that is potentially the most convincing is still in progress; also the variability in amount and chemical form of Se in plants is a problem. These gaps in understanding bioactive compounds and their health benefits should not serve to reduce research interest but should, instead, encourage plant and nutritional scientists to work together to develop strategies for improvement of health through food. PMID- 15784696 TI - Identification of genes expressed differentially in subcutaneous and visceral fat of cattle, pig, and mouse. AB - The factors that control fat deposition in adipose tissues are poorly understood. It is known that visceral adipose tissues display a range of biochemical properties that distinguish them from adipose tissues of subcutaneous origin. However, we have little information on gene expression, either in relation to fat deposition or on interspecies variation in fat deposition. The first step in this study was to identify genes expressed in fat depot of cattle using the differential display RT-PCR method. Among the transcripts identified as having differential expression in the two adipose tissues were cell division cycle 42 homolog (CDC42), prefoldin-5, decorin, phosphate carrier, 12S ribosomal RNA gene, and kelch repeat and BTB domain containing 2 (Kbtbd2). In subsequent experiments, we determined the expression levels of these latter genes in the pig and in mice fed either a control or high-fat diet to compare the regulation of fat accumulation in other animal species. The levels of CDC42 and decorin mRNA were found to be higher in visceral adipose tissue than in subcutaneous adipose tissue in cattle, pig, and mice. However, the other genes studied did not show consistent expression patterns between the two tissues in cattle, pigs, and mice. Interestingly, all genes were upregulated in subcutaneous and/or visceral adipose tissues of mice fed the high-fat diet compared with the control diet. The data presented here extend our understanding of gene expression in fat depots and provide further proof that the mechanisms of fat accumulation differ significantly between animal species. PMID- 15784695 TI - Profile of changes in gene expression in cultured hippocampal neurones evoked by the GABAB receptor agonist baclofen. AB - Metabotropic gamma-aminobutyric acid receptors (GABA(B)Rs) play a critical role in inhibitory synaptic transmission in the hippocampus. However, little is known about a possible long-term effect requiring transcriptional changes. Here, using microarray technology and RT-PCR of RNA from cultured rat embryonic hippocampal neurones, we report the profile of genes that are up- or downregulated by activation of GABA(B)Rs by baclofen but are not changed by baclofen in the presence of the GABA(B)R antagonist CGP-55845A. Our data show, for the first time, regulation of transcription of defined mRNAs after specific GABA(B) receptor activation. The identified genes can be grouped into those encoding signal transduction, endocytosis/trafficking, and structural classes of proteins. For example, butyrylcholinesterase, brain-derived neurotrophic factor, and COPS5 (Jab1) genes were upregulated, whereas Rab8 interacting protein and Rho GTPase activating protein 4 were downregulated. These results provide important baseline genomic data for future studies aimed at investigating the long-term effects of GABA(B)R activation in neurones such as their roles in neuronal growth, pathway formation and stabilization, and synaptic plasticity. PMID- 15784697 TI - Small interfering RNA-mediated functional silencing of vasopressin V2 receptors in the mouse kidney. AB - The antidiuretic effects of arginine vasopressin (AVP) on the kidney are mediated by V2 subtype AVP receptors (V2R). To investigate the role of regulation of V2R in water and sodium homeostasis, we have developed a method for small interfering RNA (siRNA)-mediated inhibition of V2R expression in vivo. Three 21-nt siRNA sequences were chosen that specifically targeted the mouse V2R but shared no appreciable sequence homology to any other known mouse genes, including the vasopressin V1a and V1b receptors. Additionally, an siRNA sequence that shared no significant matches to any known mammalian gene sequences was chosen for use as a control. Chemically synthesized siRNA was complexed with the liposomal transfection reagent DOTAP. Each mouse (male C57BL/6) received 3.6 nmol (approximately 50 microg) of either the control (nonsilencing) or one of the V2R targeting siRNAs via intravenous injection. Forty-eight hours after injection membranes were prepared from the inner medulla of the kidneys, and V2R expression was measured by a radioligand binding assay and Western immunoblotting. Treatment with one of the V2R-targeting siRNAs (R2) caused a 39.7 +/- 8.7% reduction in V2R specific binding compared with the control (n = 11, P < 0.05) and a 37.0 +/- 2.3% reduction in V2R protein expression as measured by Western immunoblotting (n = 4, P < 0.001). Additionally, real-time PCR revealed that R2 siRNA treatment induced a 68.8 +/- 2.2% reduction in V2R mRNA. However, this siRNA treatment did not alter the animals' basal urine concentrating capacity under unstimulated conditions. In subsequent experiments, treatment with R2 siRNA was found to significantly attenuate the antidiuretic effects the V2R-specific AVP agonist 1 desamino-[8-D-arginine]vasopressin (dDAVP). Mice were infused with dDAVP (0.25 ng/h) for 3 days to produce maximal antidiuresis and then were injected with either the R2 siRNA or the nonsilencing control. On day 2 after treatment, urine osmolality was significantly decreased from 3,455 +/- 72 in control animals (n = 12) to 3,155 +/- 129 mosmol/kgH2O in R2 siRNA-treated animals (n = 12) (P < 0.05); similarly, on day 2 24-h urine volume was significantly increased from 0.86 +/- 0.07 ml/day to 1.11 +/- 0.06 ml/day in R2 siRNA-treated animals (P < 0.05). In summary we have demonstrated that RNA interference methodology can be used successfully in vivo to significantly reduce functional expression of the V2R in the mouse kidney. PMID- 15784698 TI - Role of Toll-like receptor-4 in genetic susceptibility to lung injury induced by residual oil fly ash. AB - The mechanisms of susceptibility to particle-induced lung injury are not clearly understood. To evaluate the contribution of genetic background to pulmonary pathogenesis, we compared the lung injury responses to residual oil fly ash (ROFA) in inbred mouse strains and calculated heritability estimates. Significant interstrain (genetic) variation was observed in ROFA-induced lung inflammation and hyperpermeability phenotypes; broad-sense heritability ranged from approximately 0.43 to 0.62, and the coefficient of genetic determination ranged from 0.28 to 0.45. C3H/HeJ (HeJ) mice were most resistant to the ROFA-induced injury responses. This was particularly important, as HeJ mice contain a dominant negative mutation in Toll-like receptor-4 (Tlr4). We then characterized ROFA induced injury and TLR4 signaling in HeJ mice and its coisogenic strain C3H/HeOuJ (OuJ; Tlr4 normal) to understand the potential role of Tlr4 in this model. ROFA induced lung injury was significantly greater in OuJ mice compared with HeJ mice. ROFA also significantly enhanced transcript and protein levels of lung TLR4 in OuJ but not in HeJ mice. Greater activation of downstream signal molecules (i.e., MYD88, TRAF6, IRAK-1, NF-kappaB, MAPK, AP-1) was observed in OuJ mice than in HeJ mice before the development of ROFA-induced pulmonary injury. Putative TLR4 dependent inflammatory genes that were differentially induced by ROFA in the two strains include interleukin-1beta and tumor necrosis factor-alpha. Results support an important contribution of genetic background to particle-mediated lung injury, and Tlr4 is a candidate susceptibility gene. PMID- 15784699 TI - Increasing mammalian cardiomyocyte contractility with residues identified in trout troponin C. AB - The Ca2+ sensitivity of force generation in trout cardiac myocytes is significantly greater than that from mammalian hearts. One mechanism that we have suggested to be responsible, at least in part, for this high Ca2+ sensitivity is the isoform of cardiac troponin C (cTnC) found in trout hearts (ScTnC), which has greater than twice the Ca2+ affinity of mammalian cTnC (McTnC). Here, through a series of mutations, the residues in ScTnC responsible for its high Ca2+ affinity have been identified as being Asn2, Ile28, Gln29, and Asp30. When these residues in McTnC were mutated to the trout-equivalent amino acid, the Ca2+ affinity of the molecule, determined by monitoring the fluorescence of a Trp inserted for a Phe at residue 27, is comparable to that of ScTnC. To determine how a McTnC mutant containing Asn2, Ile28, Gln29, and Asp30 (NIQD McTnC) affects the Ca2+ sensitivity of force generation, endogenous cTnC in single, chemically skinned rabbit cardiomyocytes was replaced with either wild-type McTnC or NIQD McTnC. Our results demonstrate that the cardiomyocytes containing NIQD McTnC were approximately twice as sensitive to Ca2+, illustrating that a McTnC mutant with similar Ca2+ affinity as ScTnC can be used to sensitize mammalian cardiac myocytes to Ca2+. PMID- 15784700 TI - Respiratory chain defects may present only with hypoglycemia. AB - Hypoglycemia occasionally results from oxidative phosphorylation deficiency, associated with liver failure. Conversely, in some cases of respiratory chain defect, the impairment in glucose metabolism occurs with normal hepatic function. The mechanism for this hypoglycemia remains poorly understood. We report here three unrelated children with hypoglycemia as the presenting symptom associated with oxidative phosphorylation deficiency but without liver dysfunction. Two patients had, respectively, complex III and complex IV deficiency and presented with long fast hypoglycemia. During a fasting test, the first patient showed evidence for impaired gluconeogenesis (progressive increase of plasma lactate and no decrease of alanine levels), whereas the second patient appeared to have impaired fatty acid oxidation (hypoketotic hypoglycemia with increased levels of non esterified fatty acids). The third patient presented with both long and short fast hypoglycemia related to complex IV deficiency. The mechanism of hypoglycemia for this patient may have been partly related to GH insufficiency, whereas impaired glycogen metabolism possibly accounted for short fast hypoglycemia. We suggest that hypoglycemia can be the presenting symptom for respiratory chain defects, through the possible reduction in cofactors resulting from oxidative phosphorylation deficiency, and that respiratory chain defects should therefore be considered in the differential diagnosis of hypoglycemia. PMID- 15784701 TI - Inhibition of growth of experimental human endometrial cancer by an antagonist of growth hormone-releasing hormone. AB - Antagonists of GHRH are being developed for the treatment of various cancers. In this study we investigated in vivo and in vitro the effects of the GHRH antagonist MZ-J-7-118 and its mechanism of action in HEC-1A human endometrial cancer. Treatment of nude mice bearing HEC-1A xenografts with 10 mug/d MZ-J-7-118 for 6 wk significantly inhibited the volume of HEC-1A tumors by 43%, tumor weight by 40% compared with controls and prolonged the tumor doubling time from 18.7 +/- 1.4 to 25.4 +/- 3.8 d. Administration of 20 mug MZ-J-7-118, sc, twice a day significantly (P < 0.05) decreased HEC-1A growth, as evidenced by a 57.9% decrease in tumor volume, a 50.7% reduction in tumor weight, and the extension of tumor doubling time from 17.5 +/- 2.8 to 36.4 +/- 6.5 d. Therapy with GHRH antagonists significantly decreased serum IGF-I levels in experiment 1, and significantly increased tumoral IGF-I levels in experiment 2 in treated mice. Levels of IGF-II and vascular endothelial growth factor-A in tumors were not changed. Specific high affinity binding sites for GHRH were found on HEC-1A tumor membranes using ligand competition assays with (125)I-labeled GHRH antagonist JV 1-42. MZ-J-7-118 displaced radiolabeled JV-1-42 with an IC(50) of 0.13 +/- 0.04 nm. The expression of mRNA for GHRH and splice variants of the GHRH receptor in HEC-1A tumors was demonstrated by real-time RT-PCR analysis. HEC-1A cells cultured in vitro secreted GHRH into the medium. The GHRH antagonist MZ-J-7-118 inhibited the growth of HEC-1A cells in vitro. Our results indicate that GHRH antagonists can reduce the growth of human endometrial cancer and could be used as an alternative adjuvant therapy for the management of endometrial cancer. PMID- 15784702 TI - Effects of oral and transdermal estradiol administration on levels of sex hormone binding globulin in postmenopausal women with and without a history of intrahepatic cholestasis of pregnancy. AB - SHBG, the most important transport protein for sex steroids, is produced in the liver under the control of estrogen action. In a randomized, double-blind, prospective crossover study we compared basal levels of serum SHBG and their responses to increasing doses of oral and transdermal estradiol (E2), followed by E2 plus oral progestin (medroxyprogesterone acetate [MPA]), in 40 postmenopausal women with or without a history of intrahepatic cholestasis of pregnancy (ICP), which could affect the synthesis of SHBG. Serum samples collected at baseline, on the last day of each E2 period, and on the last day of the E2 plus MPA combination were assayed for SHBG and E2. Basal levels of SHBG showed no difference between the study groups. Oral but not transdermal E2 increased SHBG concentrations by 67-171% in the control group, but the response was smaller (42 121%) in the ICP group. Addition of MPA decreased SHBG levels by 14-18% in both groups during both treatments. In conclusion, a history of ICP is associated with blunted responses of SHBG to oral estrogen. PMID- 15784703 TI - The C42R mutation in the Kir6.2 (KCNJ11) gene as a cause of transient neonatal diabetes, childhood diabetes, or later-onset, apparently type 2 diabetes mellitus. AB - CONTEXT: Known genes in maturity-onset diabetes of the young account for only a fraction of families with dominantly inherited diabetes in Japan. There should be as-yet-unidentified genes that account for the rest of the patients. OBJECTIVE: To identify and characterize the mutation responsible for a Japanese family with dominantly inherited diabetes mellitus. SUBJECTS: Members of a four-generation family with dominantly inherited diabetes mellitus observed in three generations. None of the patients in this family had permanent neonatal diabetes. One had transient neonatal diabetes, one had childhood diabetes, and the others had adult onset diabetes without autoantibodies or insulin resistance. METHODS: Screening of the chromosomal location of the gene by a genome-wide linkage analysis followed by candidate gene sequencing. Confirmation of the functional significance of the identified mutation by the population survey and the physiological analysis. RESULTS: We identified a novel mutation (C42R) in the KCNJ11 gene coding for the Kir6.2 subunit of the pancreatic ATP-sensitive potassium channel. The patch-clamp experiments using the mutated KCNJ11 showed that the mutation causes increased spontaneous open probability and reduced ATP sensitivity. The effect, however, was partially compensated by the reduction of functional ATP-sensitive potassium channel expression at the cell surface, which could account for the milder phenotype of our patients. CONCLUSIONS: These results broaden the spectrum of diabetes phenotypes caused by mutations of KCNJ11 and suggest that mutations in this gene should be taken into consideration for not only permanent neonatal diabetes but also other forms of diabetes with milder phenotypes and later onset. PMID- 15784704 TI - Impact of infliximab on serum leptin levels in patients with Crohn's disease. AB - OBJECTIVES: In mice, body weight is regulated by adipocyte-derived leptin. TNFalpha is a critical mediator of inflammation-induced cachexia in Crohn's disease (CD). The regulation of leptin by TNFalpha is poorly understood in CD. Pharmacological neutralization of TNFalpha with infliximab offers a unique opportunity to study TNFalpha-mediated regulation of leptin in CD patients. METHODS: We prospectively followed up CD patients treated with infliximab (n = 20). Body composition was assessed before and after treatment at 1 and 4 wk. Serum leptin, IL-6, soluble TNF receptor type II, and soluble intercellular antiadhesion molecule-1 levels were measured as well as cholesterol levels and free urinary cortisol. Because methylprednisolone (MP) increases leptin production in vivo, CD patients treated with MP (n = 9) were studied separately as a positive control group. RESULTS: Infliximab induced clinical remission and a significant decrease in C-reactive protein (P < 0.01) and IL-6 (P < 0.05) levels in all CD patients and increased body weight (P = 0.013) at 4 wk. Leptinemia was significantly increased after infliximab administration at 1 wk (P = 0.014) and 4 wk (P < 0.001). This increase in serum leptin occurred early at 1 wk, when no significant weight and fat mass changes could be observed and was associated with the down-regulation of TNFalpha-regulated mediators, soluble TNF receptor type II (P = 0.015), and soluble intercellular antiadhesion molecule-1 (P = 0.007). Moreover, infliximab increased cholesterol levels at 1 wk (P = 0.001). Twenty four-hour cortisol secretion was not altered by infliximab. Leptinemia increased at 1 wk after MP administration (P = 0.028). CONCLUSION: Infliximab increases leptinemia in CD. This study suggests that TNFalpha exerts major inhibitory actions on leptin production in CD patients. PMID- 15784705 TI - Eight-point glucose testing versus the continuous glucose monitoring system in evaluation of glycemic control in type 1 diabetes. AB - CONTEXT: Advantages/disadvantages of continuous vs. discrete glucose monitoring are not well documented. OBJECTIVE: Compare glucose profiles from home meters vs. continuous sensors. DESIGN: Randomized clinical trial conducted by the Diabetes Research in Children Network (DirecNet) to assess the utility of the GlucoWatch G2 Biographer. SETTING: Home glucose measurements. PATIENTS: Two hundred children (age, 7 to < 18 yr) with type 1 diabetes. INTERVENTION: At baseline, subjects were asked to wear the continuous glucose monitoring system (CGMS) sensor and perform meter tests at eight prespecified times of the day (eight-point testing) each for 3 d (2 d using both, 1 d eight-point testing only, 1 d CGMS only). Hemoglobin A1c was measured in a central laboratory. MAIN OUTCOME MEASURE: Six month hemoglobin A1c. This analysis looked at baseline glucose profiles/hemoglobin A1c. RESULTS: Only 10% of subjects completed full eight-point testing for 3 d, but median CGMS use was 70 h. Mean glucose was lower when measured by the CGMS compared with eight-point testing (183 +/- 37 vs. 188 +/- 41 mg/dl; 10.2 +/- 2.1 vs.10.4 +/- 2.3 mmol/liter; P = 0.009), especially overnight (2400-0400 h; 174 vs. 199 mg/dl; 9.7 vs. 11.1 mmol/liter; P < 0.001). Associations of hemoglobin A1c with mean glucose were similar for eight-point testing [slope 23 mg/dl per 1% (1.3 mmol/liter); correlation 0.40; P < 0.001] and CGMS [slope 19 mg/dl per 1% (1.1 mmol/liter); correlation 0.39; P < 0.001]. Postprandial excursions were lower for eight-point testing vs. CGMS, especially after dinner (mean excursion -17 vs. 63 mg/dl; -1.0 vs. 3.5 mmol/liter; P < 0.001). CONCLUSIONS: Both methods gave similar mean glucose profiles and associations with hemoglobin A1c. Advantages of the CGMS were higher density of data and better detection of postprandial peaks. However, the CGMS may overestimate the frequency of low glucose levels, especially overnight. PMID- 15784706 TI - Neonatal detection of congenital hypothyroidism of central origin. AB - Due to the high frequency of concurrent pituitary hormone deficiencies, congenital hypothyroidism (CH) of central origin (CH-C) is a life-threatening disorder. Yet only a minority of these patients are detected by neonatal CH screening programs worldwide. We conducted a prospective multicenter study involving a 2-yr cohort of neonatally diagnosed CH-C patients to determine whether a T(4)-TSH-based neonatal CH screening protocol extended with T(4) binding globulin determinations improves early detection of CH-C and to assess the extent of pituitary hormone deficiency among the identified CH-C patients. In all infants with screening results indicative of CH-C, the functional integrity of the hypothalamo-hypophyseal system was investigated by dynamic tests; the anatomical integrity was investigated by magnetic resonance imaging. Initial test results were evaluated after 5 yr of follow-up. Among 385,000 infants screened over the 2-yr period, 19 cases of permanent CH-C were detected (prevalence, 1:20,263; 95% confidence interval, 1:12,976 to 1:33,654), representing 13.5% of all detected cases of permanent CH. The majority (78%) had multiple pituitary hormone deficiency, whereas 53% had pituitary malformations on magnetic resonance imaging. We conclude that infants with CH-C can very well be detected by neonatal screening. The estimated prevalence and the severity of pituitary dysfunction of this treatable disorder call for explicit attention for this entity of CH in neonatal screening programs worldwide. PMID- 15784707 TI - In vivo evidence of impaired peripheral fatty acid trapping in patients with human immunodeficiency virus-associated lipodystrophy. AB - The use of antiretroviral combination therapy in HIV has been associated with lipodystrophy and several metabolic risk factors. We postulated that patients with HIV-lipodystrophy have impaired adipose tissue free fatty acid (FFA) trapping and, consequently, increased hepatic FFA delivery. We investigated FFA, hydroxybutyric acid (HBA; reflecting hepatic FFA oxidation), and triglyceride (TG) changes after a high fat meal in HIV-infected males with (LIPO; n = 26) and without (NONLIPO; n = 12) lipodystrophy and in healthy males (n = 35). Because defective peripheral FFA trapping has been associated with impaired action of complement component 3 (C3), we also determined postprandial C3 concentrations. The LIPO group had higher homeostasis model assessment scores compared with the other groups. Areas under the curve (AUCs) for FFA, HBA, and TG were higher in the LIPO group than in the NONLIPO group or the controls. No differences in TG AUC, FFA-AUC, and HBA-AUC were observed between the NONLIPO group and controls. In HIV-infected patients, FFA-AUC and HBA-AUC were inversely related to sc adipose tissue area. Plasma C3 showed a postprandial increase in healthy controls, but not in the HIV-infected groups. C3 was not related to body fat distribution, postprandial FFA, or HBA. The present data suggest disturbed postprandial FFA metabolism in patients with HIV-lipodystrophy, most likely due to inadequate incorporation of FFA into TG in sc adipose tissue, but do not support a major role for C3 in these patients. The higher postprandial HBA levels reflect increased hepatic FFA delivery and may aggravate insulin resistance and dyslipidemia, leading to increased cardiovascular risk. PMID- 15784708 TI - Preeclampsia is associated with impaired regulation of the placental nitric oxide cyclic guanosine monophosphate pathway by corticotropin-releasing hormone (CRH) and CRH-related peptides. AB - During pregnancy, CRH and CRH-related peptides appear to regulate the fetoplacental circulation via activation of the nitric oxide (NO)/cGMP pathway. Pregnancies with abnormal placental function such as preeclampsia (PE) are characterized by increased maternal plasma CRH concentrations and reduced placental CRH-receptor 1alpha (CRH-R1alpha) expression. In this study, we investigated the actions of CRH/CRH-related peptides on the NO/cGMP system in normal and PE placentas (n = 8 for each group). Fluorescent in situ hybridization, RT-PCR, and immunofluorescence experiments in human term placenta detected mRNAs expression for both R1 and R2 types of CRH-R, as well as urocortin (UCN) II and UCN III and showed CRH-R protein expression mainly in syncytiotrophoblast, whereas the endothelial NO synthase (eNOS) expression was confined within the cytoplasm of the chorionic villi. In placental explants, CRH and UCN induced mRNA and protein expression of eNOS, but not inducible NOS, and also caused an acute increase in cGMP levels (maximal stimulation, 80-90% above basal; P < 0.05). UCN II also induced a modest induction of cGMP (42% above basal; P < 0.05). These responses were attenuated by the NOS and soluble guanylyl cyclase inhibitors, l-N(G)-nitro-l-arginine methyl ester and 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one. In PE placental explants there was a significant reduction in CRH/CRH-related peptide-induced cGMP response; however, changes in the mRNA content of eNOS, inducible NOS, and soluble guanylyl cyclase (assessed by quantitative RT-PCR) between normal and PE placentas were not altered. In conclusion, we demonstrated that CRH and CRH-related peptides can positively regulate the placental NO/cGMP system. This pathway appears to be impaired in PE and may contribute toward dysregulation of the balance controlling vascular resistance. PMID- 15784709 TI - Impact of growth hormone supplementation on adult height in turner syndrome: results of the Canadian randomized controlled trial. AB - BACKGROUND: A randomized, controlled trial of GH supplementation to adult height in girls with short stature due to Turner syndrome was conducted in Canada. We report results in subjects who completed the protocol and subjects who participated in follow-up. METHODS: One hundred fifty-four girls with Turner syndrome, aged 7-13 yr, were randomly assigned to one of two groups: 1) GH by sc injection six times per week (0.30 mg/kg.wk), and 2) control (C), no GH treatment. Both cohorts received standardized sex steroid replacement starting at a chronological age of 13 yr. Subjects were followed until protocol completion, defined as height velocity less than 2 cm/yr and bone age 14 yr or greater. A subsequent protocol addendum requested follow-up safety and efficacy assessment in all patients at least 1 yr after the last core protocol visit. RESULTS: One hundred four patients completed the study (61 GH, 43 C), and 50 withdrew (15 GH, 35 C). At protocol completion, mean heights were 147.5 +/- 6.1 (GH) and 141.0 +/- 5.4 cm (C), respectively (P < 0.001). Of those who completed the protocol, 59 (40 GH, 19 C) had height data at least 1 yr after protocol completion; in that group, mean heights were 149.0 +/- 6.4 (GH) and 142.2 +/- 6.6 cm (C), respectively (P < 0.001). At protocol completion and follow-up, the mean height gain due to GH, estimated by analysis of covariance, was +7.2 cm (confidence interval 6.0, 8.4) and +7.3 cm (confidence interval 5.4, 9.2), respectively (both P < 0.001). CONCLUSIONS: This is the first evidence from a randomized, controlled trial to adult height that GH supplementation with induction of puberty at a near physiological age increases the adult height of girls with Turner syndrome. PMID- 15784710 TI - Hypertension and hypothalamo-pituitary-adrenal axis hyperactivity affect frontal lobe integrity. AB - Chronically elevated cortisol levels have been associated with elevated blood pressure, brain atrophy, and cognitive impairments. In this cross-sectional exploratory study, we assessed whether hypertension was related to hypothalamo pituitary-adrenal axis hyperactivity and whether this may in part explain prefrontal brain atrophy and cognitive impairments in this population. We studied 27 patients with hypertension and 27 normotensive control subjects. Glucocorticoid feedback was assessed using the combined dexamethasone-CRH test. All participants completed a neuropsychological battery and received brain magnetic resonance imaging for volumetric measurement of frontal and medial temporal lobe regions. Hypertension was significantly associated with impaired glucocorticoid feedback control after statistically controlling for age, gender, and body mass index (P = 0.01). Hypertensive patients also showed a trend toward reductions in frontal lobe volume (P = 0.09) and had significantly lower scores in one of two tests of executive function (P = 0.03). Significant correlations were observed between hypothalamo-pituitary-adrenal hyperactivity and frontal lobe atrophy. Our data indicate that impaired glucocorticoid feedback control may partly account for the prefrontal volume reductions present in patients with hypertension. Future studies assessing the impact of hypertension on the brain should include cortisol assessments. PMID- 15784711 TI - A twin study for serum leptin, soluble leptin receptor, and free insulin-like growth factor-I in pubertal females. AB - CONTEXT: Leptin and IGF-I are two peripheral metabolic signals linking body energy status to hypothalamus GnRH generator and involved in the pubertal development and metabolic disorders. The changes of their biological activity through puberty and the genetic basis are not clear. OBJECTIVE: To determine the genetic and environmental influences to the variations of basal leptin, soluble leptin receptor (SOB-R), free leptin index (FLI), and free IGF-I levels in pubertal females. DESIGN: A twin study was performed in 2003. SETTING: Participants were recruited from the Qingdao Twin Registry, a school-based registry. PARTICIPANTS: A total of 360 twin girls aged 6-18 yr were enrolled, consisting of 132 pairs of monozygotic and 48 pairs of dizygotic twins. INTERVENTIONS: Anthropometric and sexual characteristics were examined. Serum total leptin and free IGF-I were measured by immunoradiometric assay, and SOB-R was measured by ELISA. MAIN OUTCOME MEASURE: Estimates of genetic and environmental components of variance were based on the theory of normal maximum likelihood in Mx package, a computer program specifically designed for the analysis of twin and family data. RESULTS: Serum leptin concentrations increased persistently throughout puberty, especially from Tanner stage III to Tanner stage IV (P < 0.05), which in consistent with the increase of percentage of body fat. However, SOB-R decreased significantly from Tanner stage I to Tanner stage II (P < 0.05), which results in a continuous rise of FLI (ratio of leptin to SOB-R), especially from Tanner stage I to Tanner stage II (P < 0.05). Serum free IGF-I increased dramatically from Tanner stage I to II and declined since then. Results of correlation analysis suggest that FLI predicts the pubertal growth and sexual maturation more effectively, whereas leptin sensitively reflects the fat mass of body composition. Quantitative genetic model fittings showed that SOB-R and free IGF-I have higher heritability (0.62-0.77, 0.54-0.66) and leptin and FLI have lower heritability (0.38-0.48, 0.44-0.55). CONCLUSIONS: Fast increase of FLI and free IGF-I from Tanner stage I to II might be involved in the onset of puberty and the onset of thelarche. The peak of free IGF-I in Tanner stage II might be presumed as an indicator of the peak of pubertal growth spurt in females, and the significant rise of leptin along with percentage of body fat from Tanner stage III to IV might be as a predictor of the forthcoming menarche. Our results stress the importance of research into the genetic regulation on the endocrine regulators involved in the pubertal development and metabolic disorders, including pubertal obesity and diabetes. PMID- 15784712 TI - Low bone formation in premenopausal women with idiopathic osteoporosis. AB - Most young people with osteoporosis have an identifiable cause. Others have an idiopathic form for which no etiology can be found. We have reported that men with idiopathic osteoporosis (IOP) have histomorphometric evidence of decreased bone formation and osteoblast dysfunction. The pathogenesis of IOP in young women remains unclear. Our aim was to characterize the histomorphometry of IOP in healthy premenopausal women. We compared iliac crest bone biopsies from nine women with IOP to 18 healthy, age-, sex-, and race-matched controls. Compared with controls, differences in bone remodeling were identified, particularly in cancellous bone. Although cancellous bone volume did not differ, there was a trend toward lower trabecular number and increased separation in women with IOP. In cancellous bone, there was no increase in osteoid width or perimeter, but IOP patients had lower bone formation parameters, including a 10% reduction in wall width (P < 0.01), an 18% reduction in mineral apposition rate (P < 0.01), and a 42% reduction in mineralized perimeter (P < or = 0.02). Additionally, the bone formation rate was 52% lower (0.026 +/- 0.004 vs. 0.054 +/- 0.01 microm/microm(2).d; P < 0.01), and a trend toward decreased activation frequency was observed in IOP patients. Conversely, bone resorption was altered in IOP patients, reflected by a longer resorption period (134 +/- 35 vs. 38 +/- 6 d; P < or = 0.02) and increased eroded perimeter (5.5 +/- 0.7 vs. 4.1 +/- 0.4%; P = 0.05). Wall width and mineralized perimeter were similarly lower in endocortical bone. Resorption period and eroded perimeter were higher in intracortical bone. Women with IOP have uncoupling of resorption and formation and, like men with IOP, osteoblast dysfunction. PMID- 15784713 TI - Periovulatory increases in tissue homing potential of circulating CD56(bright) cells are associated with fertile menstrual cycles. AB - CD56(bright) lymphocytes appear in the uterus 3-5 d after ovulation coincident with the onset of stromal cell decidualization. Although the source of these uterine immune cells is not defined, a subset of blood CD56(bright) cells exhibits enhanced capacity to adhere to decidual vascular endothelium during the periovulatory period of menstrual cycles. In this study, the effects of early pregnancy on the adhesive capacity of CD56(bright) cells to bind uterine substrates were examined in a time-course study of 18 infertile women undergoing natural cycles before transfer of frozen/thawed embryos and 18 infertile women undergoing controlled ovarian stimulation. There were three pregnancies in the natural cycle group and seven in the hormone-stimulated cohort. Hormone levels, and number and quality of transferred embryos were similar between pregnant and nonpregnant cycles. However, the adhesive function of CD56(bright) cells increased before ovulation in hormone-treated women who became pregnant and before embryo transfer in naturally cycling women who became pregnant. This pattern of incremental adhesion, which was less frequently observed in unsuccessful cycles, suggests a role for NK cells in implantation. These results support the idea that temporal control of NK cell homing to the uterine microenvironment is a prerequisite to pregnancy. PMID- 15784714 TI - Estrogen receptors and estrogen-metabolizing enzymes in human ovaries during fetal development. AB - Estrogen action plays a crucial role in many processes throughout the human life span, including development. Estrogens are pivotal in the regulation of female reproduction, but little is known about their role during ovarian development. To better understand estrogen action during ovarian development, the expression of estrogen receptors (ERs)-alpha and -beta and key enzymes regulating estradiol production, 17beta-hydroxysteroid dehydrogenases (17HSDs) types 1, 2, and 7, were analyzed in human fetal ovaries. The expression of ERs was related to the development of ovarian follicles. Before the 26th week of fetal life ERalpha was only occasionally detected, but from then onward, its expression was detected in ovarian follicles. Consistent expression of ERbeta was seen from the 20th week until term. Both ERalpha and ERbeta were localized to the granulosa cells and oocytes. Expression of 17HSD1 and 17HSD7 enzymes, catalyzing the conversion of estrone to more active estradiol, was detected as early as at the 17th week of fetal life. The expression of 17HSD1 displayed a pattern similar to that of ERs and increased toward term, whereas that of 17HSD7 decreased and was negative by the 36th week. 17HSD1 was localized to the granulosa cells, whereas 17HSD7 expression was more diffuse and was found in both granulosa and stromal cells. 17HSD2, converting estradiol to less potent estrone, was negative in all samples studied. The simultaneous appearance of estrogen-converting enzymes and ERs at the time of follicle formation indicates that the machinery for estrogen action exists in fetal ovaries and suggests a possible role for estrogens in the developing ovary. PMID- 15784715 TI - Alendronate for the treatment of osteopenia in anorexia nervosa: a randomized, double-blind, placebo-controlled trial. AB - Osteopenia is a serious medical complication of anorexia nervosa, with no known effective treatment. We conducted a double-blinded, randomized trial comparing alendronate (10 mg daily) with placebo in 32 adolescents with anorexia nervosa (mean age, 16.9 +/- 1.9 yr). All subjects received 1200 mg elemental calcium and 400 IU vitamin D daily and received the same multidisciplinary treatment for their eating disorder. Bone mineral densities (BMDs) of the lumbar spine and femoral neck were measured by dual energy x-ray absorptiometry at baseline and after 1 yr of treatment. Twenty-nine subjects completed the study. Femoral neck and lumbar spine BMDs increased 4.4 +/- 6.4% and 3.5 +/- 4.6% in the alendronate group compared with increases of 2.3 +/- 6.9% and 2.2 + 6.1% in the control group (P = 0.41, femoral neck; P = 0.53, lumbar spine). From baseline to follow-up, BMD increased significantly at the femoral neck (P = 0.02) and lumbar spine (P = 0.02) in those receiving alendronate, but did not increase in those assigned placebo (P = 0.22, femoral neck; P = 0.18, lumbar spine). At follow-up, body weight was the most important determinant of BMD. BMD was significantly higher in subjects who were weight-restored compared with those who remained at low weight (P = 0.002, femoral neck; P = 0.04, lumbar spine). After controlling for body weight, treatment group assignment still had an independent effect at the femoral neck. We conclude that in adolescents with anorexia nervosa, weight restoration is the most important determinant of BMD, but treatment with alendronate did increase the BMD of the lumbar spine and femoral neck within the group receiving alendronate, but not compared with placebo in the primary analysis. Until additional studies have demonstrated efficacy and long-term safety, the use of alendronate in this population should be confined to controlled clinical trials. PMID- 15784716 TI - Age-related pituitary volumes in prepubertal children with normal endocrine function: volumetric magnetic resonance data. AB - BACKGROUND: Evaluation of the size of the pituitary gland on magnetic resonance imaging (MRI) may be difficult, considering the wide variation in normal gland morphology. Given the paucity of age-related biometric data, our purpose was to obtain standard normal reference values for pituitary volumes in prepubertal children using three-dimensional MRI data. METHODS: Children under the age of 10 yr undergoing brain MRI for seizures or idiopathic developmental delay and who had no endocrine abnormality were recruited prospectively over 2 yr. All MRI studies included a three-dimensional sequence. Only subjects with normal studies were included. One hundred thirty-nine children were eligible (mean age, 5.2 yr). Direct pituitary volumes were measured from contiguous 1-mm thick reconstructed coronal and sagittal images. Estimated pituitary volumes were calculated using pituitary height, width, and length. RESULTS: Volumes obtained from reconstructions in either plane were essentially identical. There was a linear increase in log-transformed pituitary volume with age, but relatively weak correlations with height or body mass index. There was no gender difference and only weak correlations between pituitary height and pituitary volume and between estimated pituitary volume calculation and measured pituitary volume. We provide age-related reference ranges for pituitary volumes in graphical and tabular forms. PMID- 15784717 TI - The mitogen-activated protein kinase dependent expression of prostaglandin H synthase-2 and interleukin-8 messenger ribonucleic acid by myometrial cells: the differential effect of stretch and interleukin-1{beta}. AB - Infection and uterine stretch are the common causes of preterm labor. IL-1beta plays a key role in infection-induced preterm labor and increases prostaglandin H synthase 2 (PGHS-2) and IL-8 expression. We have shown that mechanical stretch of uterine myocytes in vitro up-regulates the expression of PGHS-2 and IL-8. In this study, we tested the hypotheses that both IL-1beta and mechanical stretch increase the myometrial expression of PGHS-2 and IL-8 via MAPK activation and that their effects are synergistic. MAPK activation was assessed in myocytes obtained from pregnant women undergoing cesarean section before the onset of labor after exposure to IL-1beta and stretch either alone or in combination. Specific inhibitors of ERK, p38, and c-Jun N-terminal kinase were used to define the role of each in the increased expression of PGHS-2 and IL-8 mRNA. We found that both IL-1beta and stretch activated all three MAPK subtypes but that they had no synergistic effect. The inhibitor studies showed that stretch-induced increases in both PGHS-2 and IL-8 mRNA expression were ERK1/2 and p38 dependent and that IL-1beta-induced increases of PGHS-2 mRNA expression were also ERK1/2 and p38 dependent, but those of IL-8 were dependent only on ERK1/2 activation. These data show that exposure of human uterine myocytes to both stretch and IL 1beta activates the MAPK system, which is responsible for the increase in PGHS-2 and IL-8 mRNA expression. We found no evidence of a synergistic effect of IL 1beta and stretch on myometrial expression of PGHS-2 and IL-8 mRNA. PMID- 15784718 TI - Short-term administration of supraphysiological recombinant human growth hormone (GH) does not increase maximum endurance exercise capacity in healthy, active young men and women with normal GH-insulin-like growth factor I axes. AB - CONTEXT: Despite the fact that the use of GH as a doping agent in sports is widespread, little is known about its short-term effects. OBJECTIVE: The objective was to study the effects of GH on exercise capacity. DESIGN: A double blind, placebo-controlled study was used, with a treatment period of 28 d. SETTING: Subjects from general community studied ambulatory at a university hospital. PARTICIPANTS: Thirty healthy active young normal volunteers (15 women and 15 men) were recruited by local announcement, and all completed the study. INTERVENTION: All subjects were randomized to receive a low GH dose (0.033 mg/kg.d or 0.1 IU/kg.d), a high GH dose (0.067 mg/kg.d or 0.2 IU/kg.d), or placebo. MAIN OUTCOME MEASURES: Power output and oxygen uptake on bicycle exercise were the main outcome measures. RESULTS: We found no effect of the low or high dosages of GH on maximum oxygen uptake during exercise (mean +/- se for placebo, 45.2 +/- 1.6 to 45.2 +/- 2.1 ml/kg.min; GH low dose, 42.8 +/- 1.6 to 42.8 +/- 1.6 ml/kg.min; GH high dose, 44.8 +/- 3.4 to 44.8 +/- 2.2 ml/kg.min; not significant by two-way ANOVA). Neither was there any effect on maximum achieved power output during exercise or on blood pressure, heart rate, or the electrocardiographic ST level at rest or during exercise. GH significantly increased total body weight (P = 0.028), an effect predominantly ascribed to fluid retention (increased extracellular water volume), whereas muscle mass (as indicated by intracellular water volume) did not change. However, changes in the latter correlated to changes in physical performance, possibly due to different training efforts. CONCLUSION: Administration of supraphysiological recombinant human GH during a period of 4 wk does not improve power output or oxygen uptake. PMID- 15784719 TI - Relaxin and prostaglandin E(2) regulate interleukin 11 during human endometrial stromal cell decidualization. AB - Decidualization of endometrial stromal cells and IL-11 signaling are essential for embryo implantation in the mouse. We investigated the effects of relaxin (RLX) and prostaglandin E(2) (PGE(2)) on IL-11 secretion by human endometrial stromal cells (HESC) and during cAMP or medroxyprogesterone acetate (P)-induced decidualization. cAMP-decidualized HESC secreted high levels of IL-11. RLX, cAMP, or PGE(2) increased IL-11 mRNA and IL-11 secretion, with maximal response to RLX and cAMP. Addition of the cAMP/protein kinase A inhibitor Rp-adenosine-3,5-cyclic monophosphorothioate to either RLX- or PGE(2)-treated cells decreased IL-11 secretion. Indomethacin treatment decreased IL-11 secretion, which was largely restored by cotreatment with PGE(2) or RLX. Cotreatment of HESC with RLX, PGE(2), or cAMP and estrogen plus P down-regulated IL-11 mRNA and IL-11 secretion at 24 h, before secretion of prolactin (decidualization marker). Addition of W147AIL-11 (IL-11 signaling inhibitor) reduced prolactin secretion stimulated by RLX or PGE(2) and estrogen plus P. This is the first demonstration that cAMP decidualized HESC secrete IL-11 and that IL-11 mRNA and IL-11 secretion are regulated by RLX and PGE(2), partly via a cAMP/protein kinase A-dependent pathway. Blocking IL-11 signaling reduced RLX+P- or PGE(2)+P-induced decidualization, suggesting that RLX and PGE(2) act via IL-11. This is important in understanding implantation and regulation of fertility. PMID- 15784720 TI - Akt/FOXO3a signaling modulates the endothelial stress response through regulation of heat shock protein 70 expression. AB - To identify new antiapoptotic targets of the PI3K-Akt signaling pathway in endothelial cells, adenovirus-mediated Akt1 gene transfer and oligonucleotide microarrays were used to examine Akt-regulated transcripts. DNA microarray analysis revealed that HSP70 expression underwent the greatest fold activation of 12,532 transcripts examined in human umbilical vein endothelial cells (HUVEC) transduced with constitutively active Akt1. Akt1 gene transfer increased HSP70 transcript expression by 24.8-fold as determined by quantitative PCR and promoted a dose-dependent up-regulation of HSP70 protein as determined by Western immunoblot analysis. Gene transfer of FOXO3a, a downstream target of Akt in endothelial cells, significantly suppressed both basal and stress-induced HSP70 protein expression. FOXO3a induced caspase-9-dependent apoptosis in HUVEC, and cotransduction with Ad-HSP70 rescued endothelial cells from FOXO3a-induced apoptosis under basal and stress conditions. Our results identify HSP70 as a new antiapoptotic target of Akt-FOXO3a signaling in endothelial cells that controls viability through modulation of the stress-induced intrinsic cell death pathway. PMID- 15784721 TI - Evidence for a functional role of the second C5a receptor C5L2. AB - During experimental sepsis in rodents after cecal ligation and puncture (CLP), excessive C5a is generated, leading to interactions with C5aR, loss of innate immune functions of neutrophils, and lethality. In the current study, we have analyzed the expression of the second C5a receptor C5L2, the putative "default" or nonsignaling receptor for C5a. Rat C5L2 was cloned, and antibody was developed to C5L2 protein. After CLP, blood neutrophils showed a reduction in C5aR followed by its restoration, while C5L2 levels gradually increased, accompanied by the appearance of mRNA for C5L2. mRNA for C5L2 increased in lung and liver during CLP. Substantially increased C5L2 protein (defined by binding of 125I-anti-C5L2 IgG) occurred in lung, liver, heart, and kidney after CLP. With the use of serum IL-6 as a marker for sepsis, infusion of anti-C5aR dramatically reduced serum IL 6 levels, while anti-C5L2 caused a nearly fourfold increase in IL-6 when compared with CLP controls treated with normal IgG. When normal blood neutrophils were stimulated in vitro with LPS and C5a, the antibodies had similar effects on release of IL-6. These data provide the first evidence for a role for C5L2 in balancing the biological responses to C5a. PMID- 15784722 TI - Identification of mTOR as a novel bifunctional target in chronic myeloid leukemia: dissection of growth-inhibitory and VEGF-suppressive effects of rapamycin in leukemic cells. AB - The mammalian target of rapamycin (mTOR) has recently been described to be constitutively activated in Bcr-Abl-transformed cells and to mediate rapamycin induced inhibition of growth in respective cell lines. We have recently shown that rapamycin down-regulates expression of vascular endothelial growth factor (VEGF), a mediator of leukemia-associated angiogenesis, in primary CML cells. In the present study, we analyzed growth-inhibitory in vitro and in vivo effects of rapamycin on primary CML cells and asked whether rapamycin-induced suppression of VEGF in leukemic cells is related to growth inhibition. Rapamycin dose dependently inhibited growth of primary CML cells obtained from patients with imatinib-responsive or imatinib-resistant disease as well as growth of Bcr-Abl transformed imatinib-resistant cell lines. Moreover, we observed potent cytoreductive effects of rapamycin in a patient with imatinib-resistant Bcr-Abl+ leukemia. The growth-inhibitory effects of rapamycin on CML cells were found to be associated with G1 cell cycle arrest and with induction of apoptosis. In all cell types tested, rapamycin was found to down-regulate expression of VEGF. However, exogenously added VEGF did not counteract the rapamycin-induced decrease in proliferation. In conclusion, rapamycin inhibits growth of CML cells in vitro and in vivo and, in addition, down-regulates expression of VEGF. Both effects may contribute to the antileukemic activity of the drug in CML. PMID- 15784723 TI - Hereditary breast/ovarian and colorectal cancer genetics knowledge in a national sample of US physicians. AB - BACKGROUND: Clinically relevant genetics knowledge is essential for appropriate assessment and management of inherited cancer risk, and for effective communication with patients. This national physician survey assessed knowledge regarding basic cancer genetics concepts early in the process of introduction of predictive genetic testing for breast/ovarian and hereditary non-polyposis colorectal cancer (HNPCC) syndromes. METHODS: A stratified random sample was selected from the American Medical Association Masterfile of all licensed physicians. In total, 1251 physicians (820 in primary care, 431 in selected subspecialties) responded to a 15 minute questionnaire (response rate 71%) in 1999-2000. Multivariate logistic regression analyses were conducted to identify demographic and practice characteristics associated with accurate response to three knowledge questions. RESULTS: Of the study population, 37.5% was aware of paternal inheritance of BRCA1/2 mutations, and 33.8% recognised that these mutations occur in <10% of breast cancer patients. Only 13.1% accurately identified HNPCC gene penetrance as >or=50%. Obstetrics/gynaecology physicians, oncologists, and general surgeons were significantly more likely than general and family practitioners to respond accurately to the breast/ovarian questions, as were gastroenterologists to the HNPCC question. CONCLUSIONS: These nationally representative data indicate limited physician knowledge about key cancer genetics concepts in 1999-2000, particularly among general primary care physicians. Specialists were more knowledgeable about syndromes they might treat or refer elsewhere. Recent dissemination of practice guidelines and continued expansion of relevant clinical literature may enhance knowledge over time. In addition to educational efforts to assist physicians with the growing knowledge base, more research is needed to characterise the organisational changes required within the healthcare system to provide effective cancer genetics services. PMID- 15784724 TI - The development of atypical haemolytic-uraemic syndrome is influenced by susceptibility factors in factor H and membrane cofactor protein: evidence from two independent cohorts. AB - BACKGROUND: In both familial and sporadic atypical haemolytic-uraemic syndrome (aHUS), mutations have been reported in regulators of the alternative complement pathway including factor H (CFH), membrane cofactor protein (MCP), and the serine protease factor I (IF). A characteristic feature of both MCP and CFH associated HUS is reduced penetrance and variable inheritance; one possible explanation for this is that functional changes in complement proteins act as modifiers. OBJECTIVE: To examine single nucleotide polymorphisms in both CFH and MCP genes in two large cohorts of HUS patients (Newcastle and Paris). RESULTS: In both cohorts there was an association with HUS for both CFH and MCP alleles. CFH and MCP haplotypes were also significantly different in HUS patients compared with controls. CONCLUSIONS: This study suggests that there are naturally occurring susceptibility factors in CFH and MCP for the development of atypical HUS. PMID- 15784725 TI - NK-dependent DC maturation is mediated by TNFalpha and IFNgamma released upon engagement of the NKp30 triggering receptor. AB - Natural killer (NK) cells were recently shown to play a relevant role in the process of dendritic cell (DC) maturation. This function is exerted either by direct DC stimulation or through killing those DCs that did not properly acquire a mature phenotype. While killing of immature DCs is dependent on the function of the NKp30 triggering receptor, the mechanism by which NK cells induce DC maturation is still unclear. In this study, we show that also the NK-mediated induction of DC maturation is dependent on NKp30. Upon NK/DC interaction, resulting in NKp30 engagement, NK cells produced tumor necrosis factor alpha (TNFalpha) (and interferon gamma [IFNgamma]) that, in turn, promoted DC maturation. Masking of NKp30 with specific monoclonal antibodies (mAbs) strongly reduced maturation of DCs cocultured with NK cells. In addition, supernatant from NK cells stimulated via NKp30 induced DC maturation, and this effect was neutralized by anti-TNFalpha antibodies (Abs). This NKp30 function is controlled by the HLA-specific inhibitory NK receptors. Accordingly, the ability to promote maturation was essentially confined to NK cells expressing the killer immunoglobulin-like receptor-negative (KIR-) NKG2A(dull) phenotype. Finally, the analysis of perforin-deficient NK cells allowed the dissection of the 2 NKp30 mediated NK-cell functions, since NKp30 could induce cytokine-dependent DC maturation in the absence of NK-mediated DC killing. PMID- 15784726 TI - Loss of Runx1 perturbs adult hematopoiesis and is associated with a myeloproliferative phenotype. AB - Homozygous loss of function of Runx1 (Runt-related transcription factor 1 gene) during murine development results in an embryonic lethal phenotype characterized by a complete lack of definitive hematopoiesis. In light of recent reports of disparate requirements for hematopoietic transcription factors during development as opposed to adult hematopoiesis, we used a conditional gene-targeting strategy to effect the loss of Runx1 function in adult mice. In contrast with the critical role of Runx1 during development, Runx1 was not essential for hematopoiesis in the adult hematopoietic compartment, though a number of significant hematopoietic abnormalities were observed. Runx1 excision had lineage-specific effects on B- and T-cell maturation and pronounced inhibition of common lymphocyte progenitor production. Runx1 excision also resulted in inefficient platelet production. Of note, Runx1-deficient mice developed a mild myeloproliferative phenotype characterized by an increase in peripheral blood neutrophils, an increase in myeloid progenitor populations, and extramedullary hematopoiesis composed of maturing myeloid and erythroid elements. These findings indicate that Runx1 deficiency has markedly different consequences during development compared with adult hematopoiesis, and they provide insight into the phenotypic manifestations of Runx1 deficiency in hematopoietic malignancies. PMID- 15784727 TI - Association of klotho, bone morphogenic protein 6, and annexin A2 polymorphisms with sickle cell osteonecrosis. AB - In patients with sickle cell disease, clinical complications including osteonecrosis can vary in frequency and severity, presumably due to the effects of genes that modify the pathophysiology initiated by the sickle mutation. Here, we examined the association of single nucleotide polymorphisms (SNPs) in candidate genes (cytokines, inflammation, oxidant stress, bone metabolism) with osteonecrosis in patients with sickle cell disease. Genotype distributions were compared between cases and controls using multiple logistic regression techniques. An initial screen and follow-up studies showed that individual SNPs and haplotypes composed of several SNPs in bone morphogenic protein 6, annexin A2, and klotho were associated with sickle cell osteonecrosis. These genes are important in bone morphology, metabolism, and vascular disease. Our results may provide insight into the pathogenesis of osteonecrosis in sickle cell disease, help identify individuals who are at high risk for osteonecrosis, and thus allow earlier and more effective therapeutic intervention. PMID- 15784728 TI - Lack of antigen-specific tissue remodeling in mice deficient in the macrophage galactose-type calcium-type lectin 1/CD301a. AB - Macrophage galactose-type C-type lectins (MGLs), which were recently named CD301, have 2 homologues in mice: MGL1 and MGL2. MGLs are expressed on macrophages and immature dendritic cells. The persistent presence of granulation tissue induced by a protein antigen was observed in wild-type mice but not in mice lacking an endogenous, macrophage-specific, galactose-type calcium-type lectin 1 (MGL1) in an air pouch model. The anti-MGL1 antibody suppressed the granulation tissue formation in wild-type mice. A large number of cells, present only in the pouch of MGL1-deficient mice, were not myeloid or lymphoid lineage cells and the number significantly declined after administration of interleukin 1 alpha (IL-1alpha) into the pouch of MGL1-deficient mice. Furthermore, granulation tissue was restored by this treatment and the cells obtained from the pouch of MGL1 deficient mice were incorporated into the granulation tissue when injected with IL-1alpha. Taken together, MGL1 expressed on a specific subpopulation of macrophages that secrete IL-1alpha was proposed to regulate specific cellular interactions crucial to granulation tissue formation. PMID- 15784729 TI - A critical function for B-Raf at multiple stages of myelopoiesis. AB - Raf kinases play an integral role in the classic mitogen-activated protein (MAP) kinase (Raf/MEK/extracellular signal-related kinase [ERK]) intracellular signaling cascade, but their role in specific developmental processes is largely unknown. Using a genetic approach, we have identified a role for B-Raf during hematopoietic progenitor cell development and during megakaryocytopoiesis. Fetal liver and in vitro embryonic stem (ES) cell-derived myeloid progenitor development is quantitatively impaired in the absence of B-Raf. Biochemical data suggest that this phenotype is due to the loss of a normally occurring rise in B Raf expression and associated ERK1/2 activation during hematopoietic progenitor cell formation. However, the presence of B-raf-/- ES cell-derived myeloid progenitors in the bone marrow of adult chimeric mice indicates the lack of an obligate cell-autonomous requirement for B-Raf in myeloid progenitor development. The lack of B-Raf also impairs megakaryocytopoiesis. Thrombopoietin (Tpo)-induced in vitro expansion of ES cell-derived megakaryocyte-lineage cells fails to occur in the absence of B-Raf. Moreover, this quantitative in vitro defect in megakaryocyte-lineage expansion is mirrored by chimeric mice data that show reduced B-raf-/- genotype contribution in megakaryocytes relative to its contribution in myeloid progenitors. Together, these data suggest that B-Raf plays a cell-autonomous role in megakaryocytopoiesis and a permissive role in myeloid progenitor development. PMID- 15784731 TI - Type 3 repeat/C-terminal domain of thrombospondin-1 triggers caspase-independent cell death through CD47/alphavbeta3 in promyelocytic leukemia NB4 cells. AB - By means of its antiangiogenic activity, thrombospondin-1 (TSP-1) exerts indirect antitumoral action on solid tumors. Here, we investigated potential antitumor action in an in vitro cell model for promyelocytic leukemia (NB4-LR1), resistant to retinoid maturation. Purified soluble TSP-1 added to cultures induced a strong dose-dependent growth inhibition and a slowly developing maturation-independent cell death. Recombinant fragments of TSP-1 allowed mapping of these activities to its type 3 repeat/C-terminal domain, features that are distinct from those of TSP 1 action on solid tumors, previously ascribed to the type 1 repeat domain. Cell death in leukemia was characterized as a caspase-independent mechanism, without DNA fragmentation, but phosphatidylserine externalization followed by membrane permeabilization. Mitochondria membrane depolarization was inherent to TSP-1 action but did not produce release of death-promoting proteins (eg, noncaspase apoptosis regulators, apoptosis-induced factor [AIF], endonuclease G, or Omi/HtrA2 or the caspase regulators, cytochrome c or second mitochondrial activator of caspase/direct inhibitor of apoptosis protein-binding protein with low isoelectric point [Smac/DIABLO]). Although detected, reactive oxygen species (ROS) production was likely not involved in the death process. Finally, receptor agonist RFYVVM and RGD peptides indicated that TSP-1 death effects are mediated by membrane receptors CD47 and alphavbeta3. These results demonstrated a new domain-specific antitumoral activity of TSP-1 on a leukemia cell line, which extends TSP-1 therapeutic potential outside the area of vascularized solid tumors. PMID- 15784730 TI - Constitutively activated FGFR3 mutants signal through PLCgamma-dependent and independent pathways for hematopoietic transformation. AB - Ectopic expression of fibroblast growth factor receptor 3 (FGFR3) associated with t(4;14) has been implicated in the pathogenesis of human multiple myeloma. Some t(4;14) patients have activating mutations of FGFR3, of which a minority are K650E (thanatophoric dysplasia type II [TDII]). To investigate the role of autophosphorylated tyrosine residues in FGFR3 signal transduction and transformation, we characterized a series of FGFR3 TDII mutants with single or multiple Y-->F substitutions. Phenylalanine substitution of Y760, essential for phospholipase Cgamma (PLCgamma) binding and activation, significantly attenuated FGFR3 TDII-mediated PLCgamma activation, as well as transformation in Ba/F3 cells and a murine bone marrow transplant leukemia model. In contrast, single substitution of Y577, Y724, or Y770 had minimal to moderate effects on TDII dependent transformation. Substitution of all 4 non-activation loop tyrosine residues significantly attenuated, but did not abolish, TDII transforming activity. Similar observations were obtained in the context of a constitutively activated fusion TEL-FGFR3 associated with t(4;12)(p16;p13) peripheral T-cell lymphomas. Moreover, 2 independent EmuSR-FGFR3 TDII transgenic mouse lines developed a pro-B-cell lymphoma, and PLCgamma was highly activated in primary lymphoma cells as assessed by tyrosine phosphorylation. These data indicate that engagement of multiple signaling pathways, including PLCgamma-dependent and PLCgamma-independent pathways, is required for full hematopoietic transformation by constitutively activated FGFR3 mutants. PMID- 15784733 TI - Bone marrow mesenchymal stem cells express a restricted set of functionally active chemokine receptors capable of promoting migration to pancreatic islets. AB - Bone marrow-derived mesenchymal stem cells (BM-MSCs) are stromal cells with the ability to proliferate and differentiate into many tissues. Although they represent powerful tools for several therapeutic settings, mechanisms regulating their migration to peripheral tissues are still unknown. Here, we report chemokine receptor expression on human BM-MSCs and their role in mediating migration to tissues. A minority of BM-MSCs (2% to 25%) expressed a restricted set of chemokine receptors (CXC receptor 4 [CXCR4], CX3C receptor 1 [CX3CR1], CXCR6, CC chemokine receptor 1 [CCR1], CCR7) and, accordingly, showed appreciable chemotactic migration in response to the chemokines CXC ligand 12 (CXCL12), CX3CL1, CXCL16, CC chemokine ligand 3 (CCL3), and CCL19. Using human pancreatic islets as an in vitro model of peripheral tissue, we showed that islet supernatants released factors able to attract BM-MSCs in vitro, and this attraction was principally mediated by CX3CL1 and CXCL12. Moreover, cells with features of BM-MSCs were detected within the pancreatic islets of mice injected with green fluorescent protein (GFP)-positive BM. A population of bona fide MSCs that also expressed CXCR4, CXCR6, CCR1, and CCR7 could be isolated from normal adult human pancreas. This study defines the chemokine receptor repertoire of human BM-MSCs that determines their migratory activity. Modulation of homing capacity may be instrumental for harnessing the therapeutic potential of BM-MSCs. PMID- 15784734 TI - The effects of iron deficiency on estradiol-induced suppression of erythropoietin induction in rats: implications of pregnancy-related anemia. AB - Pregnant women often develop anemia concomitant with the increase in serum erythropoietin levels, which are actually lower than those of nonpregnant anemic women due to the possible suppressive effect of endogenous estradiol on erythropoietin induction. The anemia, derived from hemodilution, does not act as a drive for erythropoietin induction, but iron deficiency, often observed during pregnancy, might. In order to demonstrate this, we investigated the effects of iron deficiency on estradiol-induced suppression of erythropoietin induction in rats. Single doses of estradiol suppressed hypoxia-, cobalt-, and bleeding stimulated elevation of plasma erythropoietin levels and renal erythropoietin mRNA expression. Repeated administration of estradiol at 0.1 and 1 mg/kg for 2 months induced a slight anemic trend without elevation of plasma erythropoietin. Feeding an iron-deficient diet for 2 months induced plasma erythropoietin elevation without obvious anemia, but the simultaneous repeated administration of estradiol suppressed it and reversed the iron deficiency. Plasma erythropoietin levels had distinct negative correlations with plasma iron, plasma ferritin, and iron concentrations in the organs, but not with plasma hemoglobin level. These results suggest that iron deficiency would significantly stimulate erythropoietin induction during pregnancy, although estradiol might suppress it through iron restoration. PMID- 15784732 TI - Heat shock protein 90 inhibition sensitizes acute myelogenous leukemia cells to cytarabine. AB - Previous studies demonstrated that ataxia telangiectasia mutated- and Rad3 related (ATR) kinase and its downstream target checkpoint kinase 1 (Chk1) facilitate survival of cells treated with nucleoside analogs and other replication inhibitors. Recent results also demonstrated that Chk1 is depleted when cells are treated with heat shock protein 90 (Hsp90) inhibitor 17-allylamino 17-demethoxygeldanamycin (17-AAG). The present study examined the effects of 17 AAG and its major metabolite, 17-aminogeldanamycin (17-AG), on Chk1 levels and cellular responses to cytarabine in human acute myelogenous leukemia (AML) cell lines and clinical isolates. Cytarabine, at concentrations as low as 30 nM, caused activating phosphorylation of Chk1, loss of the phosphatase Cdc25A, and S phase slowing. Conversely, treatment with 100 to 300 nM 17-AAG for 24 hours caused Chk1 depletion that was accompanied by diminished cytarabine-induced S phase accumulation, decreased Cdc25A degradation, and enhanced cytotoxicity as measured by inhibition of colony formation and induction of apoptosis. Additional studies demonstrated that small inhibitory RNA (siRNA) depletion of Chk1 also sensitized cells to cytarabine, whereas disruption of the phosphatidylinositol 3 kinase (PI3k) signaling pathway, which is also blocked by Hsp90 inhibition, did not. Collectively, these results suggest that treatment with 17-AAG might represent a means of reversing checkpoint-mediated cytarabine resistance in AML. PMID- 15784735 TI - In vivo dynamics of retinal injury and repair in the rhodopsin mutant dog model of human retinitis pigmentosa. AB - Genetic and environmental factors modify the severity of human neurodegenerations. Retinal degenerations caused by rhodopsin gene mutations show severity differences within and between families and even within regions of the same eye. Environmental light is thought to contribute to this variation. In the naturally occurring dog model of the human disorder, we found that modest light levels, as used in routine clinical practice, dramatically accelerated the neurodegeneration. Dynamics of acute retinal injury (consisting of abnormal intraretinal light scattering) were visualized in vivo in real time with high resolution optical imaging. Long term consequences included fast or slow retinal degeneration or repair of injury depending on the dose of light exposure. These experiments provide a platform to study mechanisms of neuronal injury, repair, compensation, and degeneration. The data also argue for a gene-specific clinical trial of light reduction in human rhodopsin disease. PMID- 15784736 TI - Kinetics and molecular properties of pheromone binding and release. AB - Transient kinetic studies have shown that the uptake of the pheromone (bombykol) of the silkworm moth (Bombyx mori), by its pheromone-binding protein (PBP) BmorPBP, proceeds with an "on" rate of 0.068 +/- 0.01 microM(-1).s(-1). With the high concentration of PBP in the sensillar lymph (10 mM), the half-life for the uptake of pheromone in vivo is approximately equal to 1 ms. A pH-dependent conformational change (BmorPBP(B) --> BmorPBP(A)), associated with the release of pheromone, is a first-order reaction (k = 74.1 +/- 0.32 s(-1); t(1/2), 9.3 ms). Under physiological conditions, both reactions proceed with half-life times on the order of milliseconds, as is required for odorant-oriented navigation in insects. Molecular interactions of bombykol with both native and mutated PBPs were analyzed by a novel binding assay. A recombinant protein with the native conformation (BmorPBP) showed high binding affinity (K(D) = 105 nM) at pH 7 but low affinity (K(D) = 1,600 nM) at pH 5, when tested at both low and high KCl concentrations. A protein with a C-terminal segment deleted (BmorPBPDeltaP129 V142) was found to bind bombykol at pH 7 and at pH 5 with the same affinity as the native protein at pH 7, indicating that the C-terminal segment is essential for preventing binding at low pH. Binding studies with three mutated proteins (BmorPBPW37F, BmorPBPW127F, and BmorPBPW37A) showed that replacing Trp-37 (with Phe or Ala) or Trp-127 (with Phe) did not affect the binding affinity to bombykol. Fluorescence studies shed light on the contributions of Trp-37 and Trp 127 emissions to the overall fluorescence. PMID- 15784737 TI - Hypersensitivity of DJ-1-deficient mice to 1-methyl-4-phenyl-1,2,3,6 tetrahydropyrindine (MPTP) and oxidative stress. AB - Mutations of the DJ-1 (PARK7) gene are linked to familial Parkinson's disease. We used gene targeting to generate DJ-1-deficient mice that were viable, fertile, and showed no gross anatomical or neuronal abnormalities. Dopaminergic neuron numbers in the substantia nigra and fiber densities and dopamine levels in the striatum were normal. However, DJ-1-/- mice showed hypolocomotion when subjected to amphetamine challenge and increased striatal denervation and dopaminergic neuron loss induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyrindine. DJ-1-/ embryonic cortical neurons showed increased sensitivity to oxidative, but not nonoxidative, insults. Restoration of DJ-1 expression to DJ-1-/- mice or cells via adenoviral vector delivery mitigated all phenotypes. WT mice that received adenoviral delivery of DJ-1 resisted 1-methyl-4-phenyl-1,2,3,6 tetrahydropyrindine-induced striatal damage, and neurons overexpressing DJ-1 were protected from oxidative stress in vitro. Thus, DJ-1 protects against neuronal oxidative stress, and loss of DJ-1 may lead to Parkinson's disease by conferring hypersensitivity to dopaminergic insults. PMID- 15784738 TI - DNA precursor asymmetries in mammalian tissue mitochondria and possible contribution to mutagenesis through reduced replication fidelity. AB - The mutation rate of the mammalian mitochondrial genome is higher than that of the nuclear genome. Because mitochondrial and nuclear deoxyribonucleoside triphosphate (dNTP) pools are physically distinct and because dNTP concentrations influence replication fidelity, we asked whether mitochondrial dNTP pools are asymmetric with respect to each other. We report here that the concentrations of the four dNTPs are not equal in mitochondria isolated from several tissues of both young and old rats. In particular, in most tissues examined, mitochondrial dGTP concentrations are high relative to the other dNTPs. Moreover, in the presence of the biased dNTP concentrations measured in heart and skeletal muscle, the fidelity of DNA synthesis in vitro by normally highly accurate mtDNA polymerase gamma is reduced, with error frequencies increased by as much as 3 fold, due to increased formation of template T.dGTP mismatches that are inefficiently corrected by proofreading. These data, plus some published data on specific mitochondrial mutations seen in human diseases, are consistent with the hypothesis that normal intramitochondrial dNTP pool asymmetries may contribute to spontaneous mutagenesis in the mammalian mitochondrial genome. PMID- 15784739 TI - Evolution of a large ribosomal RNA multigene family in filamentous fungi: birth and death of a concerted evolution paradigm. AB - In eukaryotes, the primary components of the ribosome are encoded by multicopy nuclear ribosomal RNA (rRNA) genes: 28/26S, 18S, 5.8S, and 5S. Copies of these genes are typically localized within tandem arrays and homogenized within a genome. As a result, nuclear rRNA gene families have become a paradigm of concerted evolution. In filamentous fungi of the subphylum Pezizomycotina, 5S rRNA genes exist as a large and dispersed multigene family, with between 50 and 100 copies per genome. To determine whether these genes defy the concerted evolution paradigm, we examined the patterns of evolution of these genes by using sequences from the complete genomes of four species. Analyses of these sequences revealed (i) multiple 5S gene types within a genome, (ii) interspecies clustering of gene types, (iii) multiple identical gene types shared among species, (iv) multiple pseudogenes within a genome, and (v) presence/absence variation of individual 5S copies in comparisons of closely related species. These results demonstrate that the 5S family in these species is characterized by birth-and death evolution under strong purifying selection. Furthermore, our results suggest that birth-and-death evolution occurs at different rates in the genera examined, and that the multiplication and movement of 5S genes across the genome are highly dynamic. As such, we hypothesize that a mechanism resembling retroposition controls 5S rRNA gene amplification, dispersal, and integration in the genomes of filamentous fungi. PMID- 15784740 TI - Defective N-acetylaspartate catabolism reduces brain acetate levels and myelin lipid synthesis in Canavan's disease. AB - Canavan's disease (CD) is a fatal, hereditary disorder of CNS development that has been linked to mutations in the gene for the enzyme aspartoacylase (ASPA) (EC 3.5.1.15). ASPA acts to hydrolyze N-acetylaspartate (NAA) into l-aspartate and acetate, but the connection between ASPA deficiency and the failure of proper CNS development is unclear. We hypothesize that one function of ASPA is to provide acetate for the increased lipid synthesis that occurs during postnatal CNS myelination. The gene encoding ASPA has been inactivated in the mouse model of CD, and here we show significant decreases in the synthesis of six classes of myelin-associated lipids, as well as reduced acetate levels, in the brains of these mice at the time of peak postnatal CNS myelination. Analysis of the lipid content of white matter from a human CD patient showed decreased cerebroside and sulfatide relative to normal white matter. These results demonstrate that myelin lipid synthesis is significantly compromised in CD and provide direct evidence that defective myelin synthesis, resulting from a deficiency of NAA-derived acetate, is involved in the pathogenesis of CD. PMID- 15784741 TI - Ca(2+)-regulated structural changes in troponin. AB - Troponin senses Ca2+ to regulate contraction in striated muscle. Structures of skeletal muscle troponin composed of TnC (the sensor), TnI (the regulator), and TnT (the link to the muscle thin filament) have been determined. The structure of troponin in the Ca(2+)-activated state features a nearly twofold symmetrical assembly of TnI and TnT subunits penetrated asymmetrically by the dumbbell-shaped TnC subunit. Ca ions are thought to regulate contraction by controlling the presentation to and withdrawal of the TnI inhibitory segment from the thin filament. Here, we show that the rigid central helix of the sensor binds the inhibitory segment of TnI in the Ca(2+)-activated state. Comparison of crystal structures of troponin in the Ca(2+)-activated state at 3.0 angstroms resolution and in the Ca(2+)-free state at 7.0 angstroms resolution shows that the long framework helices of TnI and TnT, presumed to be a Ca(2+)-independent structural domain of troponin are unchanged. Loss of Ca ions causes the rigid central helix of the sensor to collapse and to release the inhibitory segment of TnI. The inhibitory segment of TnI changes conformation from an extended loop in the presence of Ca2+ to a short alpha-helix in its absence. We also show that Anapoe, a detergent molecule, increases the contractile force of muscle fibers and binds specifically, together with the TnI switch helix, in a hydrophobic pocket of TnC upon activation by Ca ions. PMID- 15784742 TI - Ancient DNA from giant extinct lemurs confirms single origin of Malagasy primates. AB - The living Malagasy lemurs constitute a spectacular radiation of >50 species that are believed to have evolved from a common ancestor that colonized Madagascar in the early Tertiary period. Yet, at least 15 additional Malagasy primate species, some of which were relative giants, succumbed to extinction within the past 2,000 years. Their existence in Madagascar is recorded predominantly in its Holocene subfossil record. To rigorously test the hypothesis that all endemic Malagasy primates constitute a monophyletic group and to determine the evolutionary relationships among living and extinct taxa, we have conducted an ancient DNA analysis of subfossil species. A total of nine subfossil individuals from the extinct genera Palaeopropithecus and Megaladapis yielded amplifiable DNA. Phylogenetic analysis of cytochrome b sequences derived from these subfossils corroborates the monophyly of endemic Malagasy primates. Our results support the close relationship of sloth lemurs to living indriids, as has been hypothesized on morphological grounds. In contrast, Megaladapis does not show a sister-group relationship with the living genus Lepilemur. Thus, the classification of the latter in the family Megaladapidae is misleading. By correlating the geographic location of subfossil specimens with relative amplification success, we reconfirm the global trend of increased success rates of ancient DNA recovery from nontropical localities. PMID- 15784743 TI - Big brains, enhanced cognition, and response of birds to novel environments. AB - The widely held hypothesis that enlarged brains have evolved as an adaptation to cope with novel or altered environmental conditions lacks firm empirical support. Here, we test this hypothesis for a major animal group (birds) by examining whether large-brained species show higher survival than small-brained species when introduced to nonnative locations. Using a global database documenting the outcome of >600 introduction events, we confirm that avian species with larger brains, relative to their body mass, tend to be more successful at establishing themselves in novel environments. Moreover, we provide evidence that larger brains help birds respond to novel conditions by enhancing their innovation propensity rather than indirectly through noncognitive mechanisms. These findings provide strong evidence for the hypothesis that enlarged brains function, and hence may have evolved, to deal with changes in the environment. PMID- 15784744 TI - His6 tag-assisted chemical protein synthesis. AB - To make more practical the total chemical synthesis of proteins by the ligation of unprotected peptide building blocks, we have developed a method to facilitate the isolation and handling of intermediate products. The synthetic technique makes use of a His6 tag at the C terminus of the target polypeptide chain, introduced during the synthesis of the C-terminal peptide segment building block. The presence of a His6 tag enables the isolation of peptide or protein products directly from ligation reaction mixtures by Ni-NTA affinity column purification. This simple approach enables facile buffer exchange to alternate reaction conditions and is compatible with direct analytical control by protein MS of the multiple ligation steps involved in protein synthesis. We used syntheses of crambin and a modular tetratricopeptide repeat protein of 17 kDa as models to examine the utility of this affinity purification approach. The results show that His6 tag-assisted chemical protein synthesis is a useful method that substantially reduces handling losses and provides for rapid chemical protein syntheses. PMID- 15784747 TI - Inferring genetic regulatory logic from expression data. AB - MOTIVATION: High-throughput molecular genetics methods allow the collection of data about the expression of genes at different time points and under different conditions. The challenge is to infer gene regulatory interactions from these data and to get an insight into the mechanisms of genetic regulation. RESULTS: We propose a model for genetic regulatory interactions, which has a biologically motivated Boolean logic semantics, but is of a probabilistic nature, and is hence able to confront noisy biological processes and data. We propose a method for learning the model from data based on the Bayesian approach and utilizing Gibbs sampling. We tested our method with previously published data of the Saccharomyces cerevisiae cell cycle and found relations between genes consistent with biological knowledge. PMID- 15784745 TI - Target-directed proteolysis at the ribosome. AB - Target directed proteolysis allows specific processing of proteins in vivo. This method uses tobacco etch virus (TEV) NIa protease that recognizes a seven-residue consensus sequence. Because of its specificity, proteins engineered to contain a cleavage site are proteolysed, whereas other proteins remain unaffected. Therefore, this approach can be used to study the structure and function of target proteins in their natural environment within living cells. One application is the conditional inactivation of essential proteins, which is based on the concept that a target containing a recognition site can be inactivated by coexpressed TEV protease. We have previously identified one site in the secretion factor SecA that tolerated a TEV protease site insert. Coexpression of TEV protease in the cytoplasm led to incomplete cleavage and a mild secretion defect. To improve the efficiency of proteolysis, TEV protease was attached to the ribosome. We show here that cleaving SecA under these conditions is one way of increasing the efficiency of target directed proteolysis. The implications of recruiting novel biological activities to ribosomes are discussed. PMID- 15784748 TI - Pair stochastic tree adjoining grammars for aligning and predicting pseudoknot RNA structures. AB - MOTIVATION: Since the whole genome sequences of many species have been determined, computational prediction of RNA secondary structures and computational identification of those non-coding RNA regions by comparative genomics become important. Therefore, more advanced alignment methods are required. Recently, an approach of structural alignment for RNA sequences has been introduced to solve these problems. Pair hidden Markov models on tree structures (PHMMTSs) proposed by Sakakibara are efficient automata-theoretic models for structural alignment of RNA secondary structures, although PHMMTSs are incapable of handling pseudoknots. On the other hand, tree adjoining grammars (TAGs), a subclass of context-sensitive grammars, are suitable for modeling pseudoknots. Our goal is to extend PHMMTSs by incorporating TAGs to be able to handle pseudoknots. RESULTS: We propose pair stochastic TAGs (PSTAGs) for aligning and predicting RNA secondary structures including a simple type of pseudoknot which can represent most known pseudoknot structures. First, we extend PHMMTSs defined on alignment of 'trees' to PSTAGs defined on alignment of 'TAG trees' which represent derivation processes of TAGs and are functionally equivalent to derived trees of TAGs. Then, we develop an efficient dynamic programming algorithm of PSTAGs for obtaining an optimal structural alignment including pseudoknots. We implement the PSTAG algorithm and demonstrate the properties of the algorithm by using it to align and predict several small pseudoknot structures. We believe that our implemented program based on PSTAGs is the first grammar-based and practically executable software for comparative analyses of RNA pseudoknot structures, and, further, non-coding RNAs. PMID- 15784746 TI - A small-molecule inhibitor of isoprenylcysteine carboxyl methyltransferase with antitumor activity in cancer cells. AB - Many key regulatory proteins, including members of the Ras family of GTPases, are modified at their C terminus by a process termed prenylation. This processing is initiated by the addition of an isoprenoid lipid, and the proteins are further modified by a proteolytic event and methylation of the C-terminal prenylcysteine. Although the biological consequences of prenylation have been characterized extensively, the contributions of prenylcysteine methylation to the functions of the modified proteins are not well understood. This reaction is catalyzed by the enzyme isoprenylcysteine carboxyl methyltransferase (Icmt). Recent genetic disruption studies have provided strong evidence that blocking Icmt activity has profound consequences on oncogenic transformation. Here, we report the identification of a selective small-molecule inhibitor of Icmt, 2-[5-(3 methylphenyl)-1-octyl-1H-indol-3-yl]acetamide (cysmethynil). Cysmethynil treatment results in inhibition of cell growth in an Icmt-dependent fashion, demonstrating mechanism-based activity of the compound. Treatment of cancer cells with cysmethynil results in mislocalization of Ras and impaired epidermal growth factor signaling. In a human colon cancer cell line, cysmethynil treatment blocks anchorage-independent growth, and this effect is reversed by overexpression of Icmt. These findings provide a compelling rationale for development of Icmt inhibitors as another approach to anticancer drug development. PMID- 15784750 TI - Clinical peer review: description of a comprehensive model in behavioral healthcare. AB - This article describes a comprehensive model of clinical peer review that was established at a large behavioral healthcare organization serving children, adolescents, and adults with developmental disabilities. The purpose of peer review is to provide routine evaluation of practice standards that will produce the highest quality of habilitative care. The major components of peer review address (a) clinical support and administration, (b) intervention policies and procedures, (c) case consultation, (d) utilization of extraordinary procedures, (e) performance improvement projects, and (f) communication at all levels of the organization. This article discusses implementation of the model, including examples and illustrations, and factors that contribute to positive outcome. This approach to peer review represents a systems methodology that can be adopted by behavioral healthcare and human service organizations to build an in-house program of clinical quality improvement. It is a necessary first step in the large-scale implementation of behavioral services. PMID- 15784749 TI - Ovarian cancer identification based on dimensionality reduction for high throughput mass spectrometry data. AB - MOTIVATION: High-throughput and high-resolution mass spectrometry instruments are increasingly used for disease classification and therapeutic guidance. However, the analysis of immense amount of data poses considerable challenges. We have therefore developed a novel method for dimensionality reduction and tested on a published ovarian high-resolution SELDI-TOF dataset. RESULTS: We have developed a four-step strategy for data preprocessing based on: (1) binning, (2) Kolmogorov Smirnov test, (3) restriction of coefficient of variation and (4) wavelet analysis. Subsequently, support vector machines were used for classification. The developed method achieves an average sensitivity of 97.38% (sd = 0.0125) and an average specificity of 93.30% (sd = 0.0174) in 1000 independent k-fold cross validations, where k = 2, ..., 10. AVAILABILITY: The software is available for academic and non-commercial institutions. PMID- 15784751 TI - Achieving human service outcomes through competency-based training: a guide for managers. AB - During the past three decades, empirically supported strategies have been demonstrated for the training of competencies--highly specific skills and behaviors that are needed to complete a critical job task. The present article reviews several examples of competency-based training in human service programs and provides guidelines for the implementation of competency-based training by managers. Four methods for delineating a competency model are described, with recommendations for using them in combination. A sample direct support worker competency is described, and a guideline is provided for conducting competency based training on the job based on the findings of research on performance-based feedback. This guide may offer some assistance to managers of human service programs. PMID- 15784752 TI - The corporate university: a model for sustaining an expert workforce in the human services. AB - The human service industry has become a complex industry in which agencies must respond to the demands of the marketplace. To respond to these demands, agencies must develop and maintain their knowledge capital by offering an extensive array of learning opportunities related to their business goals. The corporate university, a contemporary educational model designed to maintain an expert workforce, allows agencies to meet this need effectively. PMID- 15784753 TI - The development of behaviorally based public school consultation services. AB - This article describes the development of behavioral school care consultation services to public schools within a not-for-profit community behavioral health organization. An overview of the process of behavior consultation is presented. A description of the growth of behavioral school consultation services is outlined in regard to (a) the types of consultation, including individual, school-wide, and district-wide services; (b) trainings in empirically validated assessment and interventions; and (c) the development of an internship and postdoctoral fellowship program. A review of the growth of applied research, particularly in the area of school-wide interventions and its dissemination, also is presented. PMID- 15784754 TI - Behavior-based safety and occupational risk management. AB - The behavior-based approach to managing occupational risk and preventing workplace injuries is reviewed. Unlike the typical top-down control approach to industrial safety, behavior-based safety (BBS) provides tools and procedures workers can use to take personal control of occupational risks. Strategies the author and his colleagues have been using for more than a decade to teach BBS to safety leaders and line workers are presented. In addition, a conceptual model is proposed for matching the awareness and behavior of an individual with a particular BBS intervention technique. PMID- 15784755 TI - An examination of the integrity and future of the behavior analyst certification board credentials. AB - The Behavior Analyst Certification Board (BACB) credentials behavior analysts at two levels. This article examines how well the BACB and the certifications it offers adhere to accepted professional credentialing standards. Future developments in the BACB certification process and implications for behavior analysis service delivery are also explored. PMID- 15784756 TI - The expanding role of behavior analysis and support: current status and future directions. AB - Although many of the pioneers of behavior analysis thought on a large scale and encouraged others to do so, most behavior analytic projects have remained small scale. The intent of this article is to urge the application of behavior analytic principles on a large scale. This article begins with a brief history of applied behavior analysis. It then describes some early behavior analysts who thought big and describes several examples of large-scale behavioral projects. It then shows how behavior analysis fits well with the public health model and describes how behavior analytic principles can be implemented broadly to combat public health problems. The article ends with some practical advice for behavior analysts on how to think big and speculates on the future of behavior analysis. PMID- 15784757 TI - Standardization of 3-dimensional volumes in obstetric sonography: a required step for training and automation. PMID- 15784758 TI - D. Eugene Strandness, Jr, MD, and the revolution in noninvasive vascular diagnosis. Part 2: Progression of vascular disease. PMID- 15784759 TI - A new approach to fetal echocardiography: digital casts of the fetal cardiac chambers and great vessels for detection of congenital heart disease. AB - OBJECTIVE: The purpose of this study was to describe a method of 4-dimensional (4D) reconstruction of the cardiac chambers and outflow tracts using a combination of spatiotemporal image correlation, "inversion mode," and "B-flow" imaging. METHODS: Spatiotemporal image correlation and the inversion mode were used in the examination of the volume data sets of 23 fetuses with congenital heart anomalies. A subset was also examined with B-flow imaging using the gradient light algorithm. Digital reconstructions from abnormal hearts were compared with a library obtained from fetuses without abnormalities. RESULTS: Rendered images of the 4-chamber view using the inversion mode were characterized by: (1) echogenic chambers; (2) sharp delineation of chamber contours when compared with 2-dimensional (2D) images; and (3) distinct display of the myocardium, interventricular septum, interatrial septum, and mitral and tricuspid valves as anechoic structures. Ventricular septal defects, abnormal differential insertion of the atrioventricular valves, and valve atresia were well visualized with the inversion mode. The application of inversion mode or B-flow imaging to 4D rendering of the outflow tracts resulted in "digital casts" displaying the spatial relationships between the outflow tracts as well as the connections between the great arteries and ventricular chambers. The spatial relationships and communications among cardiac structures cannot be visualized with conventional 2D ultrasonography. CONCLUSIONS: The application of spatiotemporal image correlation, inversion mode, and B-flow imaging generates information about the anatomy and pathologic characteristics of the fetal heart (digital casts) that cannot be obtained with 2D fetal echocardiography. We propose that these modalities enhance the information provided by ultrasonographic interrogation of the fetal heart and will improve prenatal diagnosis. PMID- 15784760 TI - Middle cerebral artery peak systolic velocity: technique and variability. AB - OBJECTIVE: Assessment of the middle cerebral artery (MCA) peak systolic velocity (PSV) can accurately diagnose fetal anemia and has decreased the number of invasive procedures, such as amniocentesis and cordocentesis. The objective of this investigation was to evaluate the intraobserver and interobserver variability as a measure of reproducibility of MCA PSV. The technique of correctly sampling this vessel is described. METHODS: The study population included 30 appropriate-for-gestational-age fetuses. In each fetus, MCA PSV was determined proximal to the transducer at 3 different locations: 2 mm after its origin from the internal carotid artery, at the midlength between its origin and division, and at its division. The peak systolic velocity was also determined at the contralateral MCA 2 mm after its origin. With each measurement (obtained at 2 different institutions), care was taken to ensure that the ultrasound beam was parallel to the artery for its entire length. The reliability of an angle corrector was also assessed. The intraobserver and interobserver reliabilities were determined from the appropriate version of the intraclass correlation. RESULTS: Gestational age at study entry ranged from 14 to 37.5 weeks (median, 23.6 weeks). The proximal MCA, 2 mm after its origin from the internal carotid artery, had the best intraobserver and interobserver variability in both institutions. (Intraclass correlation ranged from 0.98 to 0.99.) CONCLUSIONS: Our data indicate that fetal MCA PSV is optimally measured soon after the MCA's origin from the internal carotid artery. Given the importance of clinical decision making based on this measurement, sonographers and sonologists interested in measuring MCA PSV should test their variability after a suitable period of training. PMID- 15784761 TI - Assessment of skeletal muscle perfusion using contrast-enhanced ultrasonography. AB - OBJECTIVE: The purpose of this study was to develop a clinically applicable examination method to assess perfusion of the skeletal muscle using contrast enhanced ultrasonography (CEUS) analyzing replenishment kinetics of microbubbles. METHODS: Power Doppler sonography (7 MHz) after intravenous bolus injection of 10 mL of a microbubble contrast agent was used to repeatedly examine the perfusion of the right biceps muscle at rest and after defined exercise in 10 healthy volunteers. Parameters of perfusion, such as local blood volume, blood flow velocity, and perfusion, were calculated by a modified analysis of replenishment kinetics. For validation, CEUS was correlated with venous occlusion plethysmography (VOP) examining the right forearm flexor muscles at rest and after defined exercise. RESULTS: The CEUS examination was easily feasible and was able to depict the physiologic large variability of the right biceps muscle perfusion at rest (mean +/- SD, 3.0 +/- 2.3 [approximately mL/s x 100 mg]) compared with the results after exercise (22.9 +/- 11.0 [approximately mL/s x 100 mg]). The perfusion calculated with VOP significantly correlated with the CEUS parameters perfusion (r = 0.81; P < .001) and blood volume (r = 0.82; P < .001). The calculated mean blood flow velocity in the right forearm flexor muscles raised from 0.41 +/- 0.24 mm/s at rest to 0.64 +/- 0.39 mm/s after exercise, showing a significant correlation with the CEUS perfusion (r = 0.72; P < .001). CONCLUSIONS: Muscle perfusion can be easily and quantitatively assessed with CEUS. Compared with VOP, CEUS allows for a separate analysis of different muscle groups, unaffected by skin perfusion. Its application may be of particular interest in the diagnosis and monitoring of pathologic microvascularization in myositis or diabetic obstructive disease. PMID- 15784762 TI - On the usefulness of the mechanical index displayed on clinical ultrasound scanners for predicting contrast microbubble destruction. AB - OBJECTIVE: The purpose of this study was to evaluate the mechanical index (MI) displayed on clinical ultrasound scanners as a predictor of exposure conditions related to the destruction of sonographic microbubble contrast agents. METHODS: Sonazoid (GE Healthcare, Oslo, Norway) and Optison (GE Healthcare, Princeton, NJ) microbubbles were injected into a tissue-mimicking flow phantom. Gray scale imaging was performed with 4 different scanners and 3 different transducers (3.5 MHz curved linear, 2.5 MHz convex, and 7.5 MHz linear array), and the MI displayed by the scanner was varied from 0.2 to 1.5 by changing the system output power. All other scanning parameters were kept constant. Downstream changes in echogenicity were monitored with a PowerVision 7000 scanner (Toshiba America Medical Systems, Tustin, CA) as an indirect measure of bubble destruction. Video intensity changes within the flow tube were determined as a function of MI for the different scanner/transducer combinations, and the best linear fit was determined. RESULTS: At a displayed MI of 0.7, different scanner/transducer combinations exhibited a range in video intensity from +16% to -3% of baseline for Sonazoid and from +8% to -71% for Optison. At an MI of 0.3, reductions in video intensity of up to 32% were produced. These results indicate a wide range in bubble destruction at identical MI values. Likewise, regression analysis found no linear fits for all scanner/transducer combinations (r2 < 0.046). CONCLUSIONS: The MI displayed on clinical ultrasound scanners does not predict the degree of microbubble destruction and should not be used by itself to define exposure conditions for destruction of microbubble contrast agents. PMID- 15784763 TI - Transcranial color-coded sonography for the detection of middle cerebral artery stenosis. AB - OBJECTIVE: The purpose of this study was to validate the accuracy and criteria of transcranial color-coded sonography (TCCS) in detecting severe middle cerebral artery (MCA) stenosis. METHODS: One hundred ninety-three patients with acute ischemic cerebrovascular disease who received both TCCS and magnetic resonance angiography (MRA) examinations were evaluated. Middle cerebral artery stenosis assessed by MRA was graded as follows: grade 0, normal to mild (< 50%); grade 1, focal severe stenosis (> or = 50% and stenotic length within the M1 prebifurcation segment); and grade 2, diffuse severe stenosis (> or = 50% and stenotic length greater than the M1 prebifurcation segment). The peak systolic velocity (Vs) and mean velocity (Vm) of bilateral MCAs were obtained by TCCS. Estimates of sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated for TCCS relative to MRA. RESULTS: After 46 patients were excluded, 309 MCAs (grade 0, 77.3%; grade 1, 15.5%; and grade 2, 7.1%) were evaluated. The optimal diagnostic accuracy of TCCS for the detection of grade 1 MCA stenosis can be reached if Vs is 140 cm/s or higher or Vm is 90 cm/s or higher (sensitivity, 83.3%; specificity, 91.2%). The optimal diagnostic accuracy of grade 2 MCA stenosis can be obtained if V(s) is lower than 50 cm/s or V(s) is 140 cm/s or higher (sensitivity, 81.8%; specificity, 92.1%). In our study, none of the grade 1 but around half of the grade 2 stenosis showed a low flow velocity pattern. CONCLUSIONS: Transcranial color-coded sonography is reliable in detecting severe MCA stenosis. Except for high flow velocity, the addition of a low cutoff of normal flow velocity in our criteria not only increases the study sensitivity but also enables the identification of around half of diffuse severe MCA stenosis. PMID- 15784764 TI - Prognostic value of carotid intima-media thickness in detection of coronary atherosclerosis in patients with calcified aortic valve stenosis. AB - OBJECTIVE: Aortic stenosis (AS) coexists with coronary artery disease (CAD) in at least 30% of patients. Patients with concomitant CAD may benefit from simultaneous coronary bypass grafting. This study aimed to evaluate the prognostic value of carotid intima-media thickness (IMT) in patients with AS in assessing concomitant CAD. METHODS: Group I consisted of 33 patients (mean age +/ SD, 61.0 +/- 8.2 years; 18 men and 15 women) with AS but without CAD on angiograms. Group II consisted of 34 patients (64.4 +/- 8.0 years; 25 men and 9 women) with AS and CAD confirmed angiographically. A control group included 36 patients (61.2 +/- 4.9 years; 18 men and 18 women) with normal coronary arteries and no AS. Maximal IMT was assessed in all patients at the common carotid artery, bulb, and internal carotid artery and expressed as a mean value. RESULTS: There were no differences among the respective groups with regard to age, sex, frequency of hypertension, diabetes, and smoking habit, although patients with CAD were more often hyperlipemic (P = .038). The IMT of the common carotid artery, bulb, and internal carotid artery was significantly higher in patients with AS and CAD compared with both the control group and patients with AS only. The multivariable regression model revealed that CAD (P < .001), AS (P = .006), male sex (P = .034), age (P < .001), and diabetes mellitus (P = .047) were independent risk factors for IMT thickening. A mean IMT value of greater than 1.2 mm was predictive (sensitivity, 73.5%; specificity, 72.7%) of concomitant CAD in patients with AS. CONCLUSIONS: Intima-media thickness increases in patients with AS. The greatest IMT values are observed in patients with both AS and CAD. Patients with AS might be suspected of having CAD when the IMT value exceeds 1.2 mm. PMID- 15784765 TI - Atherosclerosis found on carotid ultrasonography is associated with atherosclerosis on coronary intravascular ultrasonography. AB - OBJECTIVE: Little has been reported on the relationship between left main coronary artery atherosclerosis and carotid ultrasonographic results. We evaluated the association between carotid and coronary atherosclerosis assessed by coronary intravascular ultrasonography (IVUS) in 45 patients. METHODS: We counted the number of plaques with intima-media thickness (IMT) greater than or equal to 1.1 mm and calculated a plaque score by summing all plaque thicknesses. With the use of IVUS, the percent plaque area was calculated at the proximal, middle, and distal sites of the left main coronary artery. The maximum percent plaque area and mean percent plaque area of the 3 sites were also calculated. Relationships among the degree of left main coronary artery atherosclerosis and carotid atherosclerosis and vascular risk factors were evaluated. RESULTS: The mean percent plaque area and maximum percent plaque area were increased in men and in patients with hypertension compared with women and those without hypertension (P < .1). Both the average of the maximum common carotid IMT and plaque number were correlated with both the mean percent plaque area and maximum percent plaque area (P < .05). Men, the presence of hypertension, and the average of the maximum common carotid IMT were correlated with both the mean percent plaque area and maximum percent plaque area by multiple linear regression analysis (P < .05). CONCLUSIONS: The average of the maximum common carotid IMT was significantly correlated with left main coronary artery atherosclerosis evaluated by IVUS. PMID- 15784766 TI - Reproducibility of duplex ultrasonography and air plethysmography used for the evaluation of chronic venous insufficiency. AB - OBJECTIVE: The purpose of this study was to determine the reproducibility of measurements on duplex ultrasonography (DU) and air plethysmography (APG) in subjects with post-thrombotic syndrome. METHODS: Duplex ultrasonography and APG were used to measure indices of lower limb venous reflux in 15 limbs with a history of deep vein thrombosis and evidence of venous insufficiency as diagnosed by ultrasonography. Three limbs were in class 0; 4 were in classes 1 to 3; and 8 were in classes 4 to 6, according to clinical, etiologic, anatomic, and pathophysiologic clinical classification. Duplex ultrasonography was performed 3 times on the same day, and venous diameter, area, peak reflux velocity, reflux flow volume, and reflux duration measurements were obtained. Air plethysmography was performed on 2 days, 7 to 10 days apart, with 1 measurement on the first day and 2 measurements on the second day. Values obtained from APG included outflow fraction, venous filling index, ejection fraction, and residual volume fraction. The measurements were performed by a vascular technologist blinded to the previous test results. One-way analysis of variance, the Student paired t test, and Bland-Altman plots were used to examine the statistical differences of the DU and APG parameters for all measurements. RESULTS: The mean coefficient of variation for within-subject measurements of all DU and APG parameters measured was less than 10%. Bland-Altman plots showed that there were no apparent trends with increasing values over a wide range for any of the DU parameters, nor were there any for the APG parameters. CONCLUSIONS: Under ideal conditions, when measured by a highly trained technologist, both DU and APG showed satisfactory reproducibility. PMID- 15784767 TI - Correlation between hypoechoic nodules on ultrasonography and benign hyperplasia in the prostatic outer gland. AB - OBJECTIVE: The purpose of this investigation was to study the correlation between hypoechoic nodules detected on ultrasonography and benign hyperplasia in the prostatic outer gland (POG) diagnosed by transrectal ultrasonography (TRUS) guided biopsies. METHODS: The TRUS-guided biopsies were performed on 472 patients suspected of having prostate cancer. Specimens obtained by biopsies were independently assessed by pathologists at 3 hospitals. RESULTS: The histologic results of 310 patients (65.68%) were benign. Focal hypoechoic nodules located in the POG were found in 240 patients (50.8%) on TRUS scans. Among them, in 22 patients (9.17% of the 240 patients with hypoechoic nodules), benign hyperplasia was found in focal hypoechoic nodules located in the POG. Focal nodules were seen as well circumscribed with an ovoid shape and smooth surface in 18 patients. CONCLUSIONS: Benign hyperplasia may sometimes originate in the POG and may appear as a hypoechoic nodule, similar to the appearance of prostate cancer. PMID- 15784768 TI - Sonographic characterization of the lumbar intervertebral disk with anatomic correlation and histopathologic findings. AB - OBJECTIVE: The purpose of this study was to describe the normal anatomy and abnormalities of the lumbar intervertebral disk with sonography in cadaveric specimens and to correlate the sonographic findings with pathologic findings. METHODS: Sonographic imaging with both 4.5- and 10-MHz linear array transducers was performed on 35 lumbar intervertebral disks in 13 human cadaveric spines. The cadaveric specimens were sectioned for anatomic and histopathologic evaluation. Findings on anatomic sections were correlated with the findings on corresponding sonographic images with respect to the sonographic appearance of disk components in 30 intervertebral disks. RESULTS: High-resolution sonography with a 10-MHz frequency transducer enabled distinction of the nucleus pulposus from the annulus fibrosus and assessment of the echogenic characteristics of these structures. Sonography showed numerous fine linear echoes in the outer portion of the intervertebral disk in 26 (87%) of 30 specimens, which corresponded to the normal concentric arrangement of the fibers in the periphery of the annulus fibrosus. Amorphous areas of low echogenicity in the inner portion of the annulus fibrosus (n = 14, 47%) correlated with degenerative changes of the disk on corresponding microscopic sections. The nucleus pulposus appeared relatively isoechoic (n = 5, 17%) or hyperechoic (n = 4, 13%) to the annulus fibrosus. In degenerative disks (n = 21, 70%), the nucleus pulposus showed decreased echogenicity, and differentiation between the nucleus pulposus and annulus fibrosus was difficult. CONCLUSIONS: High-resolution sonography is a simple imaging method that can show the normal lumbar intervertebral disk and degenerative changes in appropriate subjects. High-resolution sonography proves superior to conventional sonography for evaluation of the lumbar intervertebral disk. PMID- 15784769 TI - Tissue-mimicking liquid for use in exposimetry. AB - OBJECTIVE: Current determinations of diagnostic ultrasound exposure parameters (eg, peak rarefactional pressure and pulse intensity integral) are intended to correspond to propagation through soft tissue with a propagation speed of 1540 m/s and attenuation of 0.3 dB x cm(-1) x MHz(-1). These current measurements are made in water, which has very little attenuation, and a linear derating factor is applied to approximate 0.3 dB x cm(-1) x MHz(-1) attenuation. The fact that propagation through water as well as through soft tissue involves nonlinear propagation is not directly addressed. A better way to determine exposure parameters would be to use a liquid that has the desired tissue-mimicking properties, including a value of the nonlinearity parameter B/A representative of soft tissue. To be of practical use in the laboratory, the ultrasonic properties of this liquid must remain stable and spatially uniform for many months or years without need for periodic mixing by the user. METHODS: Fifty-two samples of fat free milk that was concentrated to one third of its original volume by ultrafiltration were created. Each sample was preserved by a different method. The speed of sound, attenuation, and nonlinearity parameter B/A of each sample were periodically monitored by narrowband through-transmission techniques. RESULTS: Six of the 52 samples remained liquid and retained acceptably stable acoustic properties over 22 months of storage at room temperature. CONCLUSIONS: Fat-free milk, concentrated via ultrafiltration and preserved in 1 of 6 different methods, has been found to be a stable tissue-mimicking liquid with acoustic properties appropriate for use in exposimetry. PMID- 15784770 TI - Ultrasonographic contrast media: has the time come in obstetrics and gynecology? AB - OBJECTIVE: The aim of this work was to review the technical aspects and clinical applications of contrast media (microbubbles and nanomolecular agents) in obstetric and gynecologic ultrasonographic imaging. METHODS: With the use of a computerized database (MEDLINE) and several Web-based search engines (Google Scholar and Copernic), relevant articles on ultrasonographic contrast media were reviewed. References cited in these articles and not obtained via the search engines were also reviewed. RESULTS: Ultrasonographic contrast media constitute a new and expanding technology. They are frequently used, for example, in adult cardiology. Extensive research in laboratory setups, animals, and human subjects has shown their safety and huge potential as an adjunctive tool in clinical practice. They increase signals returning from insonated tissues and are particularly effective as intravascular agents, enhancing color and Doppler signals, for instance. Preliminary results in tumor imaging are encouraging. The ultrasonographic contrast media permit pharmacokinetic perfusion studies, which may be of enormous clinical importance in the study of early cancer development. Targeted imaging and therapies are becoming a reality. Microbubbles have already brought a new dimension to diagnostic ultrasonographic imaging. Many authors have described the clinical value of these agents in liver, prostate, and breast imaging, among others. Newer types of media, the nanomolecules, are now emerging as the latest in imaging enhancers as well as therapeutic agent carriers. CONCLUSIONS: Although showing potential in imaging of the uterus and fallopian tubes as well as some obstetric applications, the contrast media, in particular the nanomolecules, seem to be most promising in ovarian cancer. PMID- 15784771 TI - Sonographic depiction of intratumoral vascularity with 2- and 3-dimensional color Doppler techniques. AB - OBJECTIVE: The purpose of this study was to describe patterns seen on 2- and 3 dimensional color Doppler sonographic depiction of intratumoral vessels and to correlate these patterns to histopathologic findings in an attempt to assess their clinical importance. METHODS: We conducted a retrospective analysis of 26 patients with ovarian masses and intratumoral abnormalities in whom standard 2- and 3-dimensional color Doppler sonography was performed. RESULTS: Two- and 3 dimensional color Doppler sonography depicted several patterns of vascularity within ovarian masses. These included masses with vascularity confined to the wall or loculus and those with central versus peripheral vascularity. The presence of central intratumoral vascularity had a high positive predictive value (90%) for malignancy. Conversely, the absence of intratumoral vascularity had a high negative predictive value (96%). CONCLUSIONS: Three-dimensional color Doppler sonography is helpful in depicting overall vessel density and branching patterns within an intratumoral abnormality. This technique seems to be useful in distinguishing benign from malignant ovarian masses. PMID- 15784772 TI - Primary ovarian ectopic pregnancy misdiagnosed as first-trimester missed abortion. PMID- 15784773 TI - An unusual well-circumscribed intratesticular traumatic hematoma: diagnosis and follow-up by sonography. PMID- 15784774 TI - Chronic sclerosing sialadenitis of the parotid gland: diagnosis using color Doppler sonography and sonographically guided needle biopsy. PMID- 15784775 TI - Real-time sonography of Killian-Jamieson diverticulum and its differentiation from thyroid nodules. PMID- 15784776 TI - A case of recanalized cardioembolic stroke: possible effect of transcranial color coded real-time sonography on thrombolytic therapy. PMID- 15784777 TI - Sonographic evaluation of the ventricular cardiac outflow tracts. PMID- 15784779 TI - Heart rate variability is associated with polymorphic variation in the choline transporter gene. AB - OBJECTIVE: The objective of this study was to determine whether interindividual variation in parasympathetic (cholinergic) and sympathetic (adrenergic) regulation of heart rate (as estimated by frequency components of heart rate variability [HRV]) may be accounted for, in part, by genetic variation in the choline transporter, a component of acetylcholine neurotransmission. METHODS: Resting HRV estimates of high- (HF) and low-frequency (LF) power and LF/HF ratio were determined from electrocardiogram recordings collected continuously over 5 minutes in 413 white individuals of European ancestry (49% men; aged 30-54 years [mean, 44 years]). Subjects were genotyped for a single nucleotide polymorphism (SNP) located in the 3' untranslated region of the choline transporter gene (CHT1). Frequencies of the alternate CHT1 alleles, labeled G and T, were 76% and 24%. RESULTS: Compared with GG homozygotes, participants having any T allele had greater HF power (p <.02), lower LF power (p <.02), and lower LF/HF ratios (p <.005). Relative to men, women had lower LF power (p <.001) and lower LF/HF ratios (p <.005). CONCLUSIONS: These findings show that polymorphic variation in the CHT1 gene is associated significantly with interindividual variability in HRV indices related to parasympathetic (cholinergic) activity. PMID- 15784780 TI - Effect of behavioral interventions on insulin sensitivity and atherosclerosis in the Watanabe heritable hyperlipidemic rabbit. AB - OBJECTIVE: A previous study suggested that insulin metabolic variables play a role in the progression of atherosclerosis in Watanabe heritable hyperlipidemic (WHHL) rabbits. The present study sought to determine: 1) if young, individually caged WHHLs are insulin-resistant relative to New Zealand white (NZW) rabbits and 2) whether dietary or exercise interventions can improve insulin sensitivity and slow the development of atherosclerosis in these animals. METHODS: Forty-two WHHLs were assigned to a dietary, exercise, or control condition, and 12 NZWs were used as a comparison control group. The intervention ran from 3 to 7 months of age, and all animals received an intravenous glucose tolerance test at the beginning and end of the intervention. RESULTS: WHHLs were insulin-resistant relative to NZWs at 3 months of age. Whereas the dietary intervention was effective in controlling insulin resistance, WHHLs in the exercise group without dietary restriction and the control group exhibited significant increases in insulin resistance. No intervention significantly influenced the progression of atherosclerosis. CONCLUSIONS: Young WHHLs are insulin-resistant during an early period when atherosclerosis is developing rapidly. Dietary restriction, but not exercise without weight control, is effective in controlling insulin metabolic variables in the WHHL model. Although dietary intervention can reduce cardiovascular risk factors such as insulin resistance, it is not effective in slowing the development of atherosclerosis in these genetically dyslipidemic animals. Similarly, exercise training, without dietary control, does not influence the progression of disease in WHHLs. PMID- 15784781 TI - Behavioral and emotional triggers of acute coronary syndromes: a systematic review and critique. AB - OBJECTIVE: The objective of this study was to review the evidence that behavioral and emotional factors are triggers of acute coronary syndromes. METHOD: Systematic review of the published literature from 1970 to 2004 of trigger events, defined as stimuli or activities occurring within 24 hours of the onset of acute coronary syndromes. RESULTS: There is consistent evidence that physical exertion (particularly by people who are not normally active), emotional stress, anger, and extreme excitement can trigger acute myocardial infarction and sudden cardiac death in susceptible individuals. Many triggers operate within 1 to 2 hours of symptom onset. There are methodologic limitations to the current literature, including sampling, retrospective reporting, and presentation biases, the role of memory decay and salience, and reverse causation because of silent prodromal events. CONCLUSIONS: Behavioral and emotional factors are probable triggers of acute coronary syndromes in vulnerable individuals, and the pathophysiological processes elicited by these stimuli are being increasingly understood. The benefits to patients of knowledge to these processes have yet to accrue. PMID- 15784782 TI - Inflammatory markers and sleep disturbance in major depression. AB - OBJECTIVE: This study was conducted to determine whether immune activation occurs in major depression, and to evaluate the associations between disordered sleep and markers of inflammation in patients with major depressive disorder. METHODS: All-night polysomnography was obtained in patients with acute Diagnostic and Statistical Manual of Mental Disorders, 4th edition major depressive disorder (n = 22) and age-, gender-, and body weight-matched comparison controls (n = 18). After the onset of sleep, nocturnal serum levels of interleukin-6 (IL-6), soluble intercellular adhesion molecule (sICAM), monocyte chemotactic protein (MCP-1), and IL-6 soluble receptor (IL-6sR) were sampled. RESULTS: As compared with matched controls, depressed patients showed significant (p <.05) nocturnal elevations of circulating levels of IL-6 and sICAM. Both sleep latency and rapid eye movement (REM) density had moderate correlations with IL-6 and sICAM (r's > or = 0.30). Backward regression analyses indicated that sleep latency (beta = 0.34, p <.05) and REM density (beta = 0.27, p = .09) were better predictors of IL 6 than depressive status. Similarly, sleep latency (beta = 0.27, p = .06) and REM density (beta = 0.32, p = .02) were also better predictors of sICAM. CONCLUSION: These findings support the hypothesis that sleep disturbance is associated with elevated levels of the inflammatory markers IL-6 and sICAM. This relationship was not accounted for by other confounding factors such as age and body weight. These findings suggest that the elevations in inflammatory markers found in depressive subjects may be partially the result of disturbances of sleep initiation found in this population. PMID- 15784783 TI - Depression-related hyperglycemia in type 1 diabetes: a mediational approach. AB - OBJECTIVE: Depression is linked with hyperglycemia and with an increased risk for diabetes complications, but the mechanisms underlying these relationships have not been established. In this study, we applied mediational analysis methods to determine whether the hyperglycemic effect of depression could be mediated by poor diabetes self-care. METHODS: Depression symptoms and diabetes self-care activity were assessed in a primary care sample of 188 patients with type 1 diabetes by using the Hopkins Symptom Checklist-90 (SCL-90) and the Summary of Diabetes Self-Care Activities (SDSCA). A composite score of self-care activity was formed from SDSCA ratings for diet amount, exercise, and glucose testing. Degree of hyperglycemia (level of glycosylated hemoglobin [HbA1c]), weight, insulin dose, and other clinical characteristics were obtained from electronic medical records. Ordinary least-squares regression was used to determine the effect of depression on HbA1c level controlling for weight and insulin dose. The SDSCA score was then added to the regression model to determine whether it attenuated the effect of depression symptoms on HbA1c level, thus providing suggestive evidence of mediation from these cross-sectional data. RESULTS: Depression symptoms, poor diabetes self-care, and hyperglycemia were correlated with one another in univariate analyses (p <.05). Depression symptoms were associated with higher HbA1c after controlling for weight and insulin dose (parameter estimate for depression 0.53, t = 3.6, p <.001). Inclusion of SDSCA in the model minimally attenuated the effect of depression symptoms (adjusted parameter estimate for depression 0.50, t = 3.3, p = .001). CONCLUSIONS: These findings do not support mediation of the depression-hyperglycemia relationship by diabetes self-care behavior. Other pathways, including psychophysiological mechanisms, should be investigated. PMID- 15784784 TI - Developmental heterogeneity in adolescent depressive symptoms: associations with smoking behavior. AB - OBJECTIVE: Previous research has indicated an association between smoking and depression in adolescents, although the nature of the relationship is controversial. We sought to understand this relationship better in a prospective study by investigating whether there are subpopulations of adolescents with different relationships between smoking and depressive symptoms. METHODS: Our sample was 925 adolescents attending one of five Northern Virginia high schools, grades 9 to 12. We used General Growth Mixture Modeling as our method because it allowed identification and characterization of depressive symptoms trajectories and assessment of the effects of trajectory on 12th grade smoking. We defined the binary variable 12th grade current smoking as smoking on 1 or more of the past 30 days and more than 100 cigarettes smoked in a lifetime, versus not having smoked in the past 30 days. RESULTS: We identified three trajectories: high, medium, and low depressive symptoms. For adolescents with high symptoms, 9th grade (baseline) smoking was associated with an overall deceleration of depressive symptoms, whereas for adolescents with moderate symptoms, baseline smoking was associated with an overall acceleration in depressive symptoms. Baseline smoking was not associated with rate of change in depressive symptoms for adolescents with low symptoms, nor was it associated with baseline depressive symptoms in any trajectory. CONCLUSION: These findings demonstrate that there is a relationship between smoking and depressive symptoms in adolescents, and that the relationship can vary by developmental trajectory, suggesting etiological heterogeneity. PMID- 15784785 TI - Depressive symptoms are associated with blunted cortisol stress responses in very low-income women. AB - OBJECTIVE: The purpose of this study was to examine the association between depressive symptoms and salivary cortisol responses to stress in a high-risk population of very poor Mexican women. METHODS: Adult women (N = 1109) between the ages of 18 and 44 years (mean age, 29) were identified in a house-to-house survey in low-income areas (income <20th percentile nationally) of urban Mexico. An interview containing the Spanish version of the Center for Epidemiologic Studies--Depression Scale (CES-D) was administered to all women. The naturalistic stressor was defined as the unexpected arrival of a team of researchers at the participants' homes followed by an in-depth interview and physical assessment, with saliva samples taken at time of arrival (baseline), 25 minutes, and 50 minutes after arrival. RESULTS: The mean CES-D score was 19.42 (range, 0-53). Results of hierarchical linear modeling analyses revealed no effect of depressive symptoms on baseline salivary cortisol levels. However, a significant depressive symptom by time interaction revealed that women with elevations in depressive symptoms (CES-D scores = 35) failed to exhibit a cortisol response to the stressor. In contrast, in women with lower CES-D scores, cortisol levels significantly increased in response to the stressor. CONCLUSION: Consistent with research on individuals with major depressive disorder, results of this study demonstrate that women with very high levels of depressive symptoms exhibit blunted cortisol responses to a naturalistic psychological stressor. Results also contribute to previous research by generalizing findings to a high risk, underserved population of women. PMID- 15784786 TI - Effects of treating exhaustion in angioplasty patients on new coronary events: results of the randomized Exhaustion Intervention Trial (EXIT). AB - BACKGROUND: Extreme fatigue is a common complaint in percutaneous coronary intervention (PCI) patients, and is associated with an increased risk for new cardiac events. The objective of the Exhaustion Intervention Trial (EXIT) was to determine whether a behavioral intervention on exhaustion reduces the risk of a new coronary event after PCI. METHODS AND RESULTS: Seven hundred ten consecutive patients, ages 35 to 68 years, who felt exhausted after PCI were randomized into an intervention group and a usual-care group. The intervention was based on group therapy focusing on stressors leading to exhaustion, and on support for recovery by promoting rest and making rest more efficient. One month after PCI, 50% of the patients felt exhausted. The intervention reduced the odds of remaining exhausted at 18 months by 56% in those without a previous history of coronary artery disease (CAD) (OR = 0.44; 95% CI 0.29-0.66), but had no effect on exhaustion in those with a history of CAD (OR = 0.93; 95% CI 0.56-1.55; p = .78). The intervention did not reduce the risk of a new coronary event within 2 years (RR = 1.14; 95%CI 0.82-1.57). Post-hoc analyses suggest that the effect of the intervention was limited by a positive history of CAD, the presence of a chronic, painful condition (especially rheumatism), and by opposite effects on early and late cardiac events. CONCLUSION: A behavioral intervention in PCI patients has a beneficial effect on feelings of exhaustion. It could not be demonstrated that the intervention reduces the risk of a new coronary event within 2 years. PMID- 15784787 TI - Electromagnetic hypersensitivity: a systematic review of provocation studies. AB - OBJECTIVES: The objectives of this study were to assess whether people who report hypersensitivity to weak electromagnetic fields (EMFs) are better at detecting EMF under blind or double-blind conditions than nonhypersensitive individuals, and to test whether they respond to the presence of EMF with increased symptom reporting. METHODS: An extensive systematic search was used to identify relevant blind or double-blind provocation studies. This involved searching numerous literature databases and conference proceedings, and examining the citations of reviews and included studies. The results of relevant studies were tabulated and metaanalyses were used to compare the proportions of "hypersensitive" and control participants able to discriminate active from sham EMF exposures. RESULTS: Thirty one experiments testing 725 "electromagnetically hypersensitive" participants were identified. Twenty-four of these found no evidence to support the existence of a biophysical hypersensitivity, whereas 7 reported some supporting evidence. For 2 of these 7, the same research groups subsequently tried and failed to replicate their findings. In 3 more, the positive results appear to be statistical artefacts. The final 2 studies gave mutually incompatible results. Our metaanalyses found no evidence of an improved ability to detect EMF in "hypersensitive" participants. CONCLUSIONS: The symptoms described by "electromagnetic hypersensitivity" sufferers can be severe and are sometimes disabling. However, it has proved difficult to show under blind conditions that exposure to EMF can trigger these symptoms. This suggests that "electromagnetic hypersensitivity" is unrelated to the presence of EMF, although more research into this phenomenon is required. PMID- 15784788 TI - Potentially modifiable factors associated with disability among people with diabetes. AB - OBJECTIVE: This article seeks to identify potentially modifiable factors associated with disability among people with diabetes. STUDY DESIGN AND SETTING: Among people with diabetes (N = 4357) in a large health maintenance organization, disease severity, psychologic and behavioral risk factors for disability were assessed. Disability was evaluated by the WHO Disability Assessment Scale (WHO DAS-II), the SF-36 Social Functioning scale, and days of reduced household work. RESULTS: Depression was associated with a tenfold increase in elevated WHO-DAS-II and low SF-36 Social Functioning scores, and a fourfold increase in 20+ days of reduced household work. Minor depression and the presence of three or more diabetic complications were associated with approximately a twofold increase in disability risk. Diabetic symptoms, chronic disease comorbidity, and reduced exercise were also associated with disability. CONCLUSION: Among people with diabetes, depression, diabetic complications, and exercise are potentially modifiable factors associated with disability. This suggests that integrated, biopsychosocial approaches may be needed to understand and to ameliorate disability among people with diabetes. PMID- 15784789 TI - Lifetime prevalences of physical diseases and mental disorders in young suicide victims. AB - OBJECTIVE: Evaluation was done of a possible link between psychiatric and physical illnesses in suicide victims 25 years of age or younger. METHODS: This was a follow-up study based on a 13-year database (n = 1,585) of all suicides committed during the years 1988 to 2000 in northern Finland with linkage to national hospital discharge registers. The setting for the study was the province of Oulu, located in northern Finland. Subjects comprised 202 male and 27 female suicide victims aged 25 years or less. RESULTS: Despite the young age of the suicide victims, lifetime prevalence of physical illness was about 70% in both males and females. In relation to mental disorders, female suicide victims were affected significantly more (45%) than their male counterparts (21%). About 27% of the subjects with physical illnesses had also suffered from mental disorders, but the respective proportion among those without any physical illness was only 7%. An increased prevalence of mental disorders was found in victims with diseases of the skin and subcutaneous tissues, musculoskeletal, respiratory, and digestive systems. Furthermore, increased incidences of mental disorders were also noticed in connection with injuries, poisonings, and symptoms or signs of infectious diseases. Prevalence of mental disorders in these physical disease categories varied from 25% to 44%. CONCLUSION: We recommend a greater attention to young people with physical illnesses and other symptoms in the hope that such screening may lead to an early recognition of psychiatric disorders and suicidal tendencies. PMID- 15784790 TI - Waist-to-hip ratio is positively associated with bioavailable testosterone but negatively associated with sexual desire in healthy premenopausal women. AB - OBJECTIVE: We examined whether high waist-to-hip ratio (WHR) in women is a biomarker for increased testosterone (T) and higher levels of sexual desire. METHODS: Participants were 99 healthy nonobese premenopausal women. Trait levels of mean bioavailable T were estimated from three saliva samples collected at 8:00 am, 9:30 am, and 6:00 pm to 9:00 pm, controlling for phase of the menstrual cycle. Sexual desire was measured with the Sexual Desire Inventory. RESULTS: WHR was positively, although moderately, correlated with bioavailable T. WHR, but not T, was negatively correlated with level of sexual desire. CONCLUSIONS: These results confirm that the positive association between WHR and androgens seen in clinical populations also exists in the general population of healthy adult women. However, the relationship may not be strong enough in healthy women for WHR to serve as a useful biomarker of androgen levels in sexuality studies, especially given its association with other, perhaps negatively valued, morphologic attributes. PMID- 15784791 TI - A multiple-indicator multiple-cause model for posttraumatic stress reactions: personality, coping, and maladjustment. AB - OBJECTIVE: This study aimed to develop a multiple-indicator multiple-cause model (MIMIC) to describe the relationship among posttraumatic stress (PTSD) responses, general health problems, death anxiety, personality factors, and coping strategies among community residents exposed to the technological disasters of aircraft and train crashes. MATERIALS AND METHODS: One hundred forty-eight community residents, after exposure to the aircraft or train crash, were assessed using the Impact of Event Scale, the General Health Questionnaire-28, the Death Anxiety Scale, the Eysenck Personality Questionnaire, and the Ways of Coping Checklist. The control group (n = 90) comprised members of the general public, who had not been exposed to the disasters, from another city. RESULTS: The model showed significant associations between the impact of the disaster and general health problems, which varied depending on where community residents lived in relation to the disaster site, whether they were present when the disaster occurred, and the type of disaster. The model also suggested that death anxiety was associated with type of disaster and neuroticism. The model supported the interactive model in that personality factors interacted with coping strategies in maintaining or generating PTSD and general health problems. CONCLUSIONS: After exposure to technological disasters, community residents could develop PTSD and general health problems; however, increased death anxiety was a separate psychological reaction. The interaction between certain personality traits and coping strategies was one reason for PTSD and general health problems. PMID- 15784792 TI - Insomnia in HIV infection: a systematic review of prevalence, correlates, and management. AB - OBJECTIVE: Insomnia in people with HIV and AIDS has been widely but inconsistently reported. We present the results of a systematic review of the subject. METHODS: MEDLINE, EMBASE, PSYCHLIT, and CINAHL databases were searched, and inclusion criteria were applied. The study results were then collated and described. RESULTS: Twenty-nine studies were identified, and there was wide variation in both method and quality. Insomnia was reported frequently and at all stages of HIV infection. Early reports of sleep-specific electroencephalographic changes were not confirmed. The role of immune dysregulation, virus progression, and adverse drug effects in contributing to insomnia is unclear. The presence of cognitive impairment, an AIDS-defining illness, and treatment with efavirenz were found to be significant risk factors, but the most notable association was with psychologic morbidity. There was limited evidence for the effect of specific treatments for insomnia in HIV infection. CONCLUSIONS: This review found that psychologic morbidity was a major determinant of insomnia in HIV infection. Further study would be of value in clarifying the role of other factors, as well as measuring the impact of insomnia on functioning and quality of life in this population. PMID- 15784793 TI - Socioeconomic status differences in coping with a stressful medical procedure. AB - OBJECTIVE: The objective of this study was to test the hypothesis that socioeconomic disadvantage results in adverse emotional reactions to a novel, stressful, medical examination. METHODS: Sigmoidoscopy screening for colorectal cancer was identified as a potential stressor. A subset of participants (N = 3535) from the U.K. Flexible Sigmoidoscopy Trial completed pre- and postscreening questionnaires regarding psychologic well-being. All trial participants were sent a postscreening questionnaire after 3 months (post-flexible sigmoidoscopy [FS] sample, N = 29,804), including measures of distress (the General Health Questionnaire), anxiety (State-Trait Anxiety Inventory), a single-item measure of bowel cancer worry, and a 6-item measure of positive consequences of screening. Socioeconomic status (SES) was coded from postcodes with the Townsend Index. SES differences in changes in emotional well-being over the course of screening were evaluated in the longitudinal sample. SES differences in postscreening well-being in relation to screening outcome were evaluated in the post-FS sample. RESULTS: Bowel cancer worry and anxiety were higher in lower SES groups before screening. Both reduced after screening, but there were no SES differences in the change. In the post-FS sample, there was an SES gradient in anxiety but not in distress. Lower SES groups indicated more positive reactions. There were no interactions between SES and screening outcome for any indicator of well-being. CONCLUSIONS: Lower SES was associated with worse psychologic well-being before and after screening, but lower SES participants did not show any differentially greater adverse reactions compared with higher SES participants. Moderately stressful experiences in everyday life do not necessarily more unfavorably affect those with fewer educational and economic resources. PMID- 15784794 TI - Altered cortisol response to psychologic stress in breast cancer survivors with persistent fatigue. AB - OBJECTIVE: Fatigue is one of the most common and distressing symptoms experienced by cancer patients and survivors. However, the etiology of cancer-related fatigue has not been determined. In previous studies, we have shown alterations in morning serum cortisol levels and diurnal cortisol rhythms in fatigued breast cancer survivors compared with nonfatigued control subjects. The purpose of the current study was to evaluate cortisol responses to an experimental psychologic stressor in fatigued and nonfatigued survivors. METHODS: Participants included 27 breast cancer survivors (11 fatigued, 16 nonfatigued). All had completed cancer treatment at least 3 years previously and were currently healthy with no evidence of recurrence. A standardized laboratory stressor, the Trier Social Stress Test (TSST), was administered during a 90-minute afternoon session. Saliva samples and autonomic measures (heart rate, blood pressure) were collected at 15-minute intervals throughout the session. RESULTS: Fatigued survivors showed a significantly blunted cortisol response to the stressor compared with nonfatigued survivors, controlling for depression and other potential confounds (p <.05). No differences in autonomic measures were observed. CONCLUSIONS: These results, together with our earlier findings, suggest a dysregulation in hypothalamic pituitary-adrenal (HPA) axis responsiveness among breast cancer survivors with enduring fatigue. Although the sample size was small, results suggest that attention to the HPA axis may be important for understanding cancer-related fatigue. PMID- 15784795 TI - Changes in financial strain over three years, ambulatory blood pressure, and cortisol responses to awakening. AB - OBJECTIVE: Chronic psychosocial stress has been associated cross-sectionally with ambulatory blood pressure and with salivary cortisol, but there have been few longitudinal studies of the effects of changes in chronic stress. We assessed the influence of changes in financial strain on ambulatory blood pressure and salivary cortisol. METHODS: Data were analyzed from 160 men and women age 47 to 59 years at the first assessment (T1) who repeated ambulatory monitoring 3 years later (T2). We analyzed change in financial strain as a continuous variable, and specifically compared people who did and did not report an improvement in financial strain. RESULTS: Change in financial strain was associated with change in ambulatory systolic pressure after controlling for T1 ambulatory systolic pressure, gender, socioeconomic position, age, smoking, body mass index, and T1 financial strain (p = .041). Systolic pressure at T2 was lower in the improved financial strain (121.7 +/- 11.2 mm Hg) than in the worse/no change group (125.5 +/- 11.5 mm Hg; p = .029). The corresponding diastolic pressures averaged 78.5 +/ 7.1 mm Hg and 80.7 +/- 7.9 mm Hg, respectively (p = .061). The cortisol awakening response (difference between waking and 30 minutes later) was lower (p = .048) in men who reported improved financial strain, controlling for T1 cortisol response, socioeconomic position, age, smoking, time of waking, and T1 financial strain. There were no differences in the slope of cortisol decline over the day or in evening values. CONCLUSION: These longitudinal data extend cross sectional findings in showing associations between favorable changes in chronic stress and reduced cardiovascular and neuroendocrine activation in everyday life. PMID- 15784796 TI - Basal and stimulated hypothalamic-pituitary-adrenal axis activity in patients with functional gastrointestinal disorders and healthy controls. AB - OBJECTIVE: The aim of this study was to investigate alterations of pituitary adrenal activity under both stimulated and unstimulated conditions in patients with functional gastrointestinal disorders. METHODS: Thirty subjects who fulfilled the Rome Diagnostic Criteria for either irritable bowel syndrome or nonulcer dyspepsia and 24 healthy controls took part in the study. Free salivary morning cortisol and diurnal cortisol profiles were obtained for all subjects. On a second day, a low-dose dexamethasone suppression test was applied. Additionally, in all subjects a corticotropin-releasing hormone (CRH) challenge test was performed. RESULTS: The results show attenuated unstimulated cortisol levels in patients compared with controls. After CRH challenge, blunted adrenocorticotropic hormone and cortisol responses were observed. These findings suggest lower pituitary and adrenocortical activity in patients with functional gastrointestinal disorders. CONCLUSION: The observed pituitary-adrenal reactivity in these patients is discussed as a possible consequence of lower adrenocortical activity, possibly resulting in a disinhibition of CRH in the brain. PMID- 15784797 TI - Relative-assessed psychological factors predict sedation requirement in critically ill patients. AB - OBJECTIVE: Sedation is frequently required in critically ill, mechanically ventilated patients. Sedation and analgesia requirements may vary substantially among patients. This study examined whether psychological factors predict amount of sedation requirements beyond the effects of other biomedical parameters. METHODS: This study used a prospective correlative design in an eight-bed medical intensive care unit at a tertiary university hospital. Fifty-five adult patients requiring mechanical ventilation were included. We evaluated by questionnaires three psychological factors of patients--hostility, anxiety and desire for control (DC)--as completed by patients' relatives at entry to the intensive care unit. Daily doses of sedatives required were monitored. The primary outcome measurement was midazolam dose expressed in mg/kg/h. RESULTS: There was a statistically significant correlation between psychological factors and midazolam dose (mg/kg/h): r values = 0.40 for anxiety, 0.43 for hostility, and 0.46 for DC. Age and pulmonary edema were inversely related to midazolam requirements, whereas smoking, chronic obstructive pulmonary disease, fentanyl dose, and therapeutic intervention scoring system were positively correlated with midazolam doses. In a multiple regression, DC accounted for an additional and significant 5.4% of the variance in midazolam after controlling statistically for the effects of the significant background and biomedical predictors. In the final regression equation, DC and fentanyl were the only significant factors associated with higher sedation requirement. CONCLUSION: Premorbid psychological profile independently predicts sedation requirement in critically ill, mechanically ventilated patients. Early identification of such a profile may help in sedation management and patient care. The possible mechanisms and clinical implications are discussed. PMID- 15784798 TI - The placebo response in the treatment of chronic fatigue syndrome: a systematic review and meta-analysis. AB - OBJECTIVE: The placebo response is conventionally asserted to be high in chronic fatigue syndrome (CFS) because of the latter's subjective nature and obscure pathogenesis, but no systematic review of placebo responses has been undertaken. We report such a study. Patient expectation is known to be important in the placebo response. It is also known that CFS patients attending specialist clinics often have strong physical attributions regarding causation and hence skepticism about psychological or psychiatric interventions. If so, the placebo response in CFS may be influenced by the type of intervention according to its perceived rationale. We aimed to estimate the summary placebo response in clinical trials of CFS and to determine whether intervention type influences the placebo response in CFS. METHODS: We searched Medline, Embase, Cochrane Library, PsychInfo, and the references of the identified articles, and contacted experts for controlled trials (randomized or nonrandomized) of any intervention on CFS patients reporting the placebo response as a clinical improvement in physical or general outcomes. Data were extracted from the articles and validity assessment conducted by one reviewer and checked by a second. Meta-analysis and metaregression were performed. RESULTS: The pooled placebo response was 19.6% (95% confidence interval, 15.4-23.7), lower than predicted and lower than in some other medical conditions. The meta-regression revealed that intervention type significantly contributed to the heterogeneity of placebo response (p = .03). CONCLUSION: In contrast with the conventional wisdom, the placebo response in CFS is low. Psychological-psychiatric interventions were shown to have a lower placebo response, perhaps linked to patient expectations. PMID- 15784799 TI - The antitussive effect of placebo treatment on cough associated with acute upper respiratory infection. AB - OBJECTIVE: The objective of this study was to determine the effects of a placebo treatment on cough in patients with cough associated with acute upper respiratory tract infection (URTI). METHODS: Patients with dry or slightly productive cough associated with a history of URTI were recruited. Cough frequency (CF) over 15 minutes was recorded by means of a microphone connected to a pen recorder. Cough suppression time (CST) was recorded when patients were instructed by means of a red light to try not to cough. Patients received either a single dose of vitamin E (placebo treatment) or no treatment. CF and CST were recorded before and 15 minutes after treatment. RESULTS: Twenty-seven patients were randomized to placebo treatment and 27 to the no-treatment group (mean age 22.6 years). The median difference between post- and pretreatment CF was -3 in the no-treatment group and -18 in the placebo group (p = .0003). There was a significant increase in CST in the placebo group compared with no treatment (p = .027). CONCLUSIONS: The results demonstrate that placebo treatment has significant antitussive activity. This placebo effect may be related to generation of central neurotransmitters such as endogenous opioids. PMID- 15784800 TI - Symptom profile of multiple chemical sensitivity in actual life. AB - OBJECTIVE: This study was conducted to confirm the definition of multiple chemical sensitivity (MCS) in actual life: that multiple symptoms are provoked in multiple organs by exposure to, and ameliorated by avoidance of, multiple chemicals at low levels. We used the Ecological Momentary Assessment to monitor everyday symptoms and the active sampling and passive sampling methods to measure environmental chemical exposure. METHODS: Eighteen patients with MCS, diagnosed according to the 1999 consensus criteria, and 12 healthy controls participated in this study. Fourteen patients and 12 controls underwent 1-week measurement of physical and psychologic symptoms and of the levels of exposure to various chemicals. Linear mixed models were used to test the hypotheses regarding the symptom profile of MCS patients. RESULTS: Some causative chemicals were detected in 11 of 14 MCS patients. Two other patients did not report any hypersensitivity episodes, whereas passive sampling showed far less exposure to chemicals than control subjects. Another subject reported episodic symptoms but was excluded from the following analyses because no possible chemical was detected. Eleven of the 17 physical symptoms and all four mood subscales examined were significantly aggravated in the interview based on "patient-initiated symptom prompts." On the other hand, there were no differences in physical symptoms or mood subscales between MCS patients and control subjects in the interview based on "random prompts." CONCLUSIONS: MCS patients do not have either somatic or psychologic symptoms under chemical-free conditions, and symptoms may be provoked only when exposed to chemicals. PMID- 15784801 TI - Written emotional expression produces health benefits in fibromyalgia patients. AB - OBJECTIVE: Written expression of traumatic experiences, an intervention found to have health benefits in rheumatoid arthritis, asthma, and breast cancer, was tested in a randomized, controlled trial with female fibromyalgia patients. It was hypothesized that relative to controls, patients engaging in the writing intervention would experience improved status on psychological well-being and physical health variables. METHODS: Patients (N = 92) were randomized into a trauma writing group, a control writing group, or usual care control group. The two writing groups wrote in the laboratory for 20 minutes on 3 days at 1-week intervals. Psychological well-being, pain, and fatigue were the primary outcome variables. Assessments were made at pretreatment, posttreatment, 4-month follow up, and 10-month follow-up. RESULTS: The trauma writing group experienced significant reductions in pain (effect size [ES] = 0.49) and fatigue (ES = 0.62) and better psychological well-being (ES = 0.47) at the 4-month follow-up relative to the control groups. Benefits were not maintained at the 10-month follow-up. CONCLUSION: Fibromyalgia patients experienced short-term benefits in psychological and health variables through emotional expression of personal traumatic experiences. PMID- 15784802 TI - Latent inhibition of rotation chair-induced nausea in healthy male and female volunteers. AB - OBJECTIVES: Pre-exposure to an environment in which a nausea-inducing body rotation will subsequently be given constitutes a latent inhibition procedure that might act to reduce anticipatory and postrotation nausea. METHODS: This was tested in 24 healthy subjects randomly assigned to receive no pre-exposure (group 0), a single pre-exposure (group 1), or three pre-exposures (group 3). Rotation was standardized as 5 x 1 minute rotation, but the subjects could terminate it on request. Nausea was determined on a 7-item symptom rating scale before, during, and after rotation on days 3 and 4, whereas anticipatory nausea was measured before presumed rotation on day 5. Saliva cortisol and tumor necrosis factor alpha (TNF-alpha) levels were determined at baseline before, directly, and 15 and 30 minutes after rotation every day, and before presumed rotation on day 5. RESULTS: Pre-exposure significantly reduced the degree of anticipatory nausea on day 5. Cortisol levels increased with rotation and were higher at baseline on days 4 and 5, but subjects habituated from day 3 to day 4; levels were lower in women than in men. In contrast, TNF-alpha decreased with rotation but showed no habituation. For both cortisol and TNF-alpha, no effects on postrotational nausea were found. CONCLUSION: It is concluded that repetitive pre-exposure (latent inhibition) reduces anticipatory but not postrotation nausea; behavioral measures (rotation time) and measures of acute stress (cortisol, TNF-alpha) do not respond to latent inhibition. Thus, Pavlovian conditioning rules are effective in healthy humans with anticipatory nausea but not with postrotation nausea. Hormonal responses--TNF-alpha decrease with stress, compensatory cortisol increase--and gender-related effects on learning and habituation are discussed with regard to psychophysiological and psychoimmunological processes. PMID- 15784804 TI - Use of heroin to cope with stress caused by a negative life event in a patient with lupus erythematosus. PMID- 15784805 TI - Ramoplanin: a lipoglycodepsipeptide antibiotic. AB - OBJECTIVE: To review the pharmacology, antimicrobial activity, pharmacokinetics, clinical applications, and safety of ramoplanin, a lipoglycodepsipeptide antibiotic. DATA SOURCES: Information was obtained from MEDLINE and BIOSIS databases (1984-August 2004) and Oscient Pharmaceuticals using the key words ramoplanin, A 16686, A 16686A, and MDL 62198. STUDY SELECTION AND DATA EXTRACTION: Available English-based articles and abstracts were reviewed, along with information from Oscient Pharmaceuticals. DATA SYNTHESIS: Ramoplanin exerts its bactericidal activity against gram-positive aerobic and anaerobic bacteria by blocking peptidoglycan synthesis via lipid II. In vitro susceptibility reports have demonstrated efficacy against antibiotic-resistant gram-positive pathogens. Cross-resistance has not been documented with vancomyin and other glycopeptides. Clinical trials are investigating ramoplanin's oral administration for treatment of Clostridium difficile-associated diarrhea. Previous clinical trials had evaluated the suppression of colonization of vancomycin-resistant Enterococcus with ramoplanin. Adverse effects are minimal, and drug-drug interactions have not been documented. CONCLUSIONS: The completion of clinical trials will determine whether ramoplanin has a promising role as a treatment option for diarrhea due to C. difficile. PMID- 15784806 TI - Recombinant factor VIIa in the treatment of non-hemophiliac bleeding. AB - OBJECTIVE: To review the clinical evidence for the use of recombinant factor VIIa (rFVIIa) in the prevention and/or treatment of bleeding in non-hemophiliac patients. DATA SOURCES: A MEDLINE search (1966-December 2004) was conducted to identify pertinent literature. Results were limited to English-language reports and clinical trials. References of relevant articles and selected abstracts presented at scientific meetings were also reviewed. STUDY SELECTION AND DATA EXTRACTION: Human data from prospective and retrospective studies that examined the hemostatic effect of rFVIIa in non-hemophiliac patients were reviewed, with a focus on surgical prophylaxis, liver disease, intractable bleeding associated with trauma and surgery, and anticoagulation reversal. DATA SYNTHESIS: Results from limited controlled trials on the use of rFVIIa as an adjunct for prevention of bleeding in surgery and liver diseases have not been consistent. For treatment of intractable bleeding, earlier use of rFVIIa in one trauma trial was shown to decrease the number of blood transfusions, but no differences in terms of clinical outcomes were observed in all trials. Controlled trials do not suggest an increased risk of thrombotic events. Optimal dosing and timing of administration have yet to be defined. CONCLUSIONS: Until further prospective controlled data are available, it is recommended that conventional intervention for prevention and control of hemorrhage in non-hemophiliac patients should remain the standard of care. Close monitoring of coagulation parameters is recommended before, during, and after therapy, especially in high-risk patients. Pharmacoeconomic analysis may be useful to help control costs and maximize clinical benefits. PMID- 15784807 TI - Successful desensitization to oxaliplatin. AB - OBJECTIVE: To report the successful desensitization of a patient to oxaliplatin utilizing an 8-hour desensitization regimen in a controlled environment. CASE SUMMARY: A 53-year-old white woman with metastatic colon cancer was receiving oxaliplatin, bevacizumab, and capecitabine every 2 weeks, with a partial response to therapy. On her fifth cycle of this regimen, she experienced diaphoresis, hypotension, nausea, abdominal cramping, and coryza. According to the Naranjo probability scale, oxaliplatin, and not bevacizumab, was the probable cause of the hypersensitivity reaction. The woman continued therapy with capecitabine and bevacizumab, resulting in stable disease. Due to her initial response to the oxaliplatin-based regimen, it was decided to attempt desensitization to oxaliplatin in a controlled, inpatient environment. An 8-hour desensitization schedule was employed, and the patient successfully completed an additional 3 cycles with full-dose oxaliplatin. DISCUSSION: Hypersensitivity reactions to platinum-containing compounds are well described and potentially life threatening. With expanded use of oxaliplatin in various malignancies, an increased number of hypersensitivity reactions will likely be reported. Patients with previous hypersensitivity reactions to carboplatin are at risk for similar reactions to oxaliplatin. We achieved successful desensitization for oxaliplatin using increased concentrations of the drug over an 8-hour period concomitant with oral and intravenous corticosteroids and histamine blockers. CONCLUSIONS: Hypersensitivity reactions to platinum compounds may result in discontinuation of active therapies in patients with metastatic disease. Desensitization to oxaliplatin is possible utilizing this approach. PMID- 15784808 TI - Antidepressant use during pregnancy and the rates of spontaneous abortions: a meta-analysis. AB - BACKGROUND: Due to the high prevalence of depression in women of childbearing age and coupled with the fact that approximately 50% of the pregnancies are unplanned, there is a high chance that these women have been exposed to antidepressants in early pregnancy. OBJECTIVE: To determine baseline rates of spontaneous abortions (SAs) and whether antidepressants increase those rates. METHODS: Rates of SAs in women taking antidepressants compared with non-depressed women were combined into a relative risk using a random effects model. MEDLINE, EMBASE, Healthstar, Toxline, Psychlit, Cochrane database, and Reprotox were searched for studies published in any language from 1966 to 2003. Key words used to identify articles included pregnancy outcome, abortion, miscarriage, spontaneous, antidepressant, depression, and the generic names of each antidepressant and class. Bibliographies, review articles, and reference lists from studies were also used to identify potential articles expected to provide evidence of safety of antidepressants in pregnancy. RESULTS: Of 15 potential articles, 6 cohort studies of 3567 women (1534 exposed, 2033 nonexposed) provided extractable data. All matched on important confounders. Tests found no heterogeneity (chi2 3.13; p = 0.98), and all quality scores were adequate (>50%). The baseline SA rate (95% CI) was 8.7% (7.5% to 9.9%; n = 2033). For antidepressants, the rate was 12.4% (10.8% to 14.1%; n = 1534), significantly increased by 3.9% (1.9% to 6.0%); RR was 1.45 (1.19 to 1.77; n = 3567). No differences were found among antidepressant classes. CONCLUSIONS: Maternal exposure to antidepressants may be associated with increased risk for SA; however, depression itself cannot be ruled out. PMID- 15784810 TI - The gift of hope: my personal experience. AB - SCIENTIFIC EDITOR'S INTRODUCTORY COMMENT: As part of our conference, we began with a panel of cancer survivors recounting their personal stories of cancer, infertility, and paths to parenthood. Lindsay Nohr Beck not only had a cancer treatment that could potentially impair fertility but, in a wonderful example of "turning lemons into lemonade," founded the advocacy group Fertile Hope so that other men and women facing infertility would have a national resource to find information and support. We asked her to share her story with readers of this monograph. For more information, visit www.fertilehope.org or call (888) 994 HOPE. PMID- 15784811 TI - Motivation for parenthood after cancer: a review. AB - Research on the psychosocial aspects of parenthood after cancer is just beginning. Because of delayed childbearing and increasing success of cancer treatment, more young adults are experiencing infertility related to their past cancer treatment. Pilot surveys of 132 young men and women in one sample and 201 men in a second indicate that for a majority of cancer survivors, their illness increases the value they place on family ties. Nevertheless, 17% of women had unrealistically high anxiety about pregnancy causing cancer recurrence, and even greater percentages of survivors feared that their children would be at high risk for birth defects or cancer. More research is needed with large and diverse samples of cancer survivors to create educational and supportive interventions for those interested in becoming parents after cancer. PMID- 15784812 TI - Mechanisms of toxic damage to spermatogenesis. AB - Azoospermia and long-lasting testicular atrophy are common adverse consequences of cancer treatment. Chemotherapeutic agents may disrupt spermatogenesis by targeting various testicular cell types (Leydig cells, Sertoli cells, and germ cells) and by activating numerous molecular pathways involved in germ cell life and-death decision making. Genetically modified animal models with deficiencies in specific proapoptotic and prosurvival pathways have become powerful tools in understanding the molecular regulation of spermatogenesis and the response of the seminiferous epithelium to toxic injury. In this brief review, selected examples of results of toxic exposures in genetically deficient animal models are discussed to highlight the roles of p53 and the Fas system as modulators of proapoptotic activity in the testis. A final section focuses on cisplatin, a cancer chemotherapeutic agent that produces male reproductive toxicity by targeting multiple cell types in the testis. PMID- 15784813 TI - Disruption of spermatogenesis by the cancer disease process. AB - In the past, cancer survivors tended to be most concerned about disease recurrence and treatment side effects. As survival rates have increased, however, patients are now also concerned about quality-of-life issues such as preserving fertility potential. It is well known that cancer treatment adversely affects male fertility via direct effects on the testis and/or through the endocrine glands. Evidence also suggests that the disease process itself may affect a man's fertility by influencing spermatogenesis. However, the causes of poor semen quality in cancer patients are not well understood. Multiple factors are likely involved, including preexisting defects in germ cells, systemic effects of cancer, and endocrine and immunological disturbances. This paper will summarize available evidence on different factors involved in impaired spermatogenesis in patients with various cancers with emphasis on testicular cancer and Hodgkin lymphoma. Cryopreservation of spermatozoa is a simple and practical approach available to all patients with cancer who wish to preserve their fertilizing potential before cancer therapy. PMID- 15784814 TI - Spermatogenesis after cancer treatment: damage and recovery. AB - Treatment with cytotoxic chemotherapy and radiotherapy is associated with significant gonadal damage in men, and alkylating agents are the most common agents implicated. The vast majority of men receiving procarbazine-containing regimens for the treatment of lymphomas are rendered permanently infertile, whereas treatment with doxorubicin hydrochloride (Adriamycin), bleomycin, vinblastine, and dacarbazine appears to have a significant advantage, with a return to normal fertility in the vast majority of patients. Cisplatin-based chemotherapy for testicular cancer results in temporary azoospermia in most men, with a recovery of spermatogenesis in about 50% of the patients after 2 years and 80% after 5 years. The germinal epithelium is very sensitive to radiation-induced damage, with changes to spermatogonia following as little as 0.2 Gy. Testicular doses of less than 0.2 Gy had no significant effect on FSH levels or sperm counts, whereas doses between 0.2 and 0.7 Gy caused a transient dose-dependent increase in FSH and reduction in sperm concentration, with a return to normal values within 12-24 months. No radiation dose threshold has been defined above which permanent azoospermia is inevitable; however, doses of 1.2 Gy and above are likely to be associated with a reduced risk of recovery of spermatogenesis; the time to recovery, if it is to occur, is also likely to be dose dependent. PMID- 15784815 TI - Anti-Mullerian hormone: its role in follicular growth initiation and survival and as an ovarian reserve marker. AB - In this paper the role in the ovary of anti-Mullerian hormone (AMH), a member of the transforming growth factor-beta family of growth and differentiation factors, is reviewed. AMH has an inhibitory effect on primordial follicle recruitment and may also inhibit follicle-stimulating homone-dependent selection of follicles for dominance. In addition to its functional role in the ovary, AMH in serum is an excellent candidate marker as an indication of the ovarian reserve, not only in infertility clinic patients but also in women during and after cancer treatment. PMID- 15784816 TI - Appraisal of chemotherapy effects on reproductive outcome according to animal studies and clinical data. AB - Cancer in women or men during reproductive life raises fears and dilemmas regarding the ability to have a healthy child. Chemotherapy and radiotherapy increase genetic defects in germ cells, depending on the agent used and the stage of gamete maturation. No increase in miscarriage or congenital malformation rates is detected among children born years post-cancer treatment. However, when pregnancy occurred shortly after treatment, increased abortion and malformation risk was reported. Until more data are available, monitoring of chromosomal aberrations and birth defects is recommended. Complexity of cancer treatment is significantly amplified in women exposed to chemotherapy during pregnancy due to concerns regarding direct maternal risks caused by treatment and risk to the developing embryo-fetus. The potential teratogenic effect of cancer treatment depends upon the developmental stage of the fetus at exposure and on drugs used. During the first trimester, abortion and malformation rates are increased, while second- and third-trimester chemotherapy may increase the risk of stillbirth, fetal growth restriction, and premature birth. Maternal myelosuppression increases bleeding and infection tendency, which can harm the fetus. A multidisciplinary team alerted to possible consequences of cancer treatment on pregnancy outcome should provide the optimal treatment options for these patients. PMID- 15784817 TI - Maintenance of ovarian function and risk of premature menopause related to cancer treatment. AB - Ovarian damage following cancer therapy is dependent on age at treatment as well as the type of therapeutic exposures. Older age and exposure to higher doses of alkylating agents and higher doses of radiation to the ovary are associated with a greater likelihood of ovarian failure. Acute loss of ovarian function during or shortly following treatment is relatively uncommon in females treated during childhood and adolescence but can be seen following myeloablative, alkylator based cytoreduction (e.g., busulfan and cyclophosphamide) for stem cell transplant and following direct ovarian radiation with doses >10 Gy. For survivors who retain normal ovarian function after cancer therapy, there is an increased risk of premature menopause later in life. The risk factors associated with an early menopause include exposure to high doses of alkylating agents and abdomino-pelvic radiation. PMID- 15784818 TI - Impact of paternal exposure to chemotherapy on offspring in the rat. AB - Paternal exposure to chemotherapeutics may have adverse effects on offspring. In the rat, chronic low-dose paternal exposure to the anticancer drug cyclophosphamide increased pre- and postimplantation loss and malformations. The effects of paternal drug treatment on progeny were influenced by the stage when germ cells were first exposed. Chronic cyclophosphamide treatment resulted in a dramatic decrease in the expression of stress response genes in pachytene spermatocytes and round spermatids but not in elongated spermatids; reduced gene expression may allow damage to accumulate. Exposure for 9 weeks, but not for 6, increased the incidence of aneuploidy. DNA strand breaks were maximal 3 weeks after short-term or acute treatment, during spermiogenesis. Cyclophosphamide exposed spermatozoa imparted DNA damage to the fertilized embryo. Total RNA synthesis was higher in one-cell embryos sired by drug-treated fathers than in controls, and the expression of specific genes was altered. Thus, in the rat, paternal exposure to an anticancer drug altered germ cell quality, disrupting embryo development and dysregulating zygotic gene activation. PMID- 15784819 TI - Relative susceptibilities of male germ cells to genetic defects induced by cancer chemotherapies. AB - Some chemotherapy regimens include agents that are mutagenic or clastogenic in model systems. This raises concerns that cancer survivors who were treated before or during their reproductive years may be at increased risks for abnormal reproductive outcomes. However, the available data from offspring of cancer survivors are limited, representing diverse cancers, therapies, time to pregnancies, and reproductive outcomes. Rodent breeding data after paternal exposures to individual chemotherapeutic agents illustrate the complexity of factors that influence the risk for transmitted genetic damage including agent, dose, end point, and germ cell susceptibility profiles that vary across agents. Direct measurements of chromosomal abnormalities in sperm of mice and humans by sperm fluorescent in situ hybridization have corroborated the differences in germ cell susceptibilities. The available evidence indicates that the risk of producing chromosomally defective sperm is highest during the first few weeks after the end of chemotherapy and decays with time. Thus, sperm samples provided immediately after the initiation of cancer therapies may contain treatment induced genetic defects that will jeopardize the genetic health of offspring. PMID- 15784820 TI - Hormonal approaches to preservation and restoration of male fertility after cancer treatment. AB - It is important to develop methods to prevent or reverse the sterility caused by chemotherapy or radiation therapy for cancer in men. Using a rat model, we have shown that infertility after testicular exposure to moderate doses of radiation and some chemotherapeutic agents occurs as a result of the inability of spermatogonia to differentiate. There is evidence that this phenomenon also occurs in men. Spermatogenesis and fertility can be restored in rats following treatment with radiation or some chemotherapeutic agents by suppressing testosterone with gonadotropin releasing hormone (GnRH) agonists or antagonists either before or after the cytotoxic insult. However, species differences exist in the testicular response to radiation or GnRH antagonist therapy, so rescue protocols that work in rodents do not work in nonhuman primates. The applicability of this procedure to humans is still largely unknown. In rodents, suppression of testosterone with GnRH analog treatment also appears to enhance the success of spermatogonial transplantation-an option when all stem cells are killed by cytotoxic therapy. PMID- 15784821 TI - Preservation of fertility and ovarian function and minimization of chemotherapy induced gonadotoxicity in young women by GnRH-a. AB - Improved long-term survival in young women with lymphoma and leukemia has increased attention to the preservation of their future fertility. We have attempted to minimize the gonadotoxic effect of chemotherapy by cotreatment with a GnRH agonist analog, inducing a temporary prepubertal milieu. Our prospective clinical case series includes 92 women with lymphoma, aged 15-40 years, 10 with leukemia and 18 undergoing chemotherapeutic treatments for nonmalignant autoimmune diseases. Depot D-TRP6-GnRH-a was injected monthly from before the initiation of chemotherapy until its conclusion, for up to 6 months. We used 82 similarly treated patients with lymphoma not given GnRH-a as a comparison group. All but five of the surviving evaluable patients with GnRH-a/chemotherapy cotreatment resumed spontaneous ovulation and menses or conceived, whereas 53% of the patients in the comparison group experienced premature ovarian failure (P<.01). Mechanisms to explain this apparent chemoprotective effect are discussed, and the work of other investigators in this area is reviewed. PMID- 15784822 TI - Fertility-sparing surgery for malignancies in women. AB - Never before have women with newly diagnosed gynecologic malignancies had more options for preservation of fertility. Girls or women of childbearing age with several ovarian cancer subtypes have a high probability of unilateral ovarian involvement, and, thus, may be candidates for fertility-sparing surgery with preservation of a contralateral normal ovary and uterus. These subtypes include ovarian tumors of low malignant potential, malignant ovarian germ cell tumors, and ovarian sex cord-stromal tumors. For women with invasive epithelial ovarian cancer who have early-stage disease, fertility-sparing surgery may be an option. In some cases, fertility-sparing surgery may be followed by postoperative chemotherapy. For women with invasive cervical cancer, fertility-sparing surgery may be possible. Options include conization alone for stage IA1 or IA2 disease, radical trachelectomy with stage IA2 or IB disease, or ovarian transposition for women undergoing chemoradiation. Non-operative options, such as hormonal therapy, may be considered for women with early-stage, low-grade endometrial cancer. For all women of childbearing age with gynecologic malignancies, in vitro fertilization techniques or cryopreservation of ovarian tissue may be an option prior to definitive treatment. PMID- 15784823 TI - Treatment options for the infertile male with cancer. AB - The detrimental effects of cancer on male fertility are multifactorial and may be secondary to the disease process itself or to therapeutic interventions. Patients and their doctors need to be better informed about the options men with cancer have to preserve their fertility status. Sperm banking is a safe and effective means of storing sperm before chemotherapy, radiotherapy, or surgery. Advances in assisted reproductive techniques allow in vitro fertilization to occur with the injection of a single sperm into an egg (IVF-ICSI); consequently, the requirement for the amount of sperm needed from cryopreservation is minimal. Pregnancies have been achieved using IVF-ICSI with cryopreserved sperm from patients with a history of malignancy. Similarly, successful sperm retrieval using testicular sperm extraction combined with IVF-ICSI can be accomplished for men with nonobstructive azoospermia after chemotherapy. New techniques such as spermatogonial cell transplantation offer the potential to restore fertility in patients who have received radiation or chemotherapy. PMID- 15784824 TI - Cryopreservation and transplantation of spermatogonia and testicular tissue for preservation of male fertility. AB - The existence of spermatogonial stem cells in the testis offers clinically relevant options for preservation and restoration of male fertility. New approaches based on male germ cell transplantation and testicular tissue grafting can be applied to generate a limited number of sperm cells and could therefore be considered important new avenues for restoration of fertility in oncological patients. We have developed approaches to infuse germ cells into rodent and primate testes and shown that germ cell transplantation is a procedure for restoration of spermatogenesis in the testis that might be adaptable to primates. As a promising alternative, grafting of testicular tissue has been used to produce fertile sperm. The rapid progress in the development of novel experimental strategies to generate sperm by transplantation of spermatogonial stem cells or by grafting of testicular tissue should stimulate oncologists to consider the cryopreservation of testicular tissue. This review introduces the reader to the physiology of spermatogonial stem cells and summarizes the current and potential future options for fertility preservation in male oncological patients. PMID- 15784825 TI - Fertility preservation: a comprehensive approach to the young woman with cancer. AB - Modern treatments for cancers of the reproductive age are yielding ever-higher cure rates, but more often than not, the price paid for survival is the loss of reproductive function from gonadal toxicity. Alkylating agents and ionizing radiation have well-recognized deleterious effects within the testes and ovary and cause sterility in a high proportion of patients exposed to these treatments. Preservation of fertility for men simply involves the banking of sperm before treatment, but for women, the storage of gametes is technically very complex and has limited success. Even when faced with the diagnosis of cancer, many reproductive-aged women are burdened by the possibility of never conceiving a child with their own eggs. Fertility preservation for the reproductive-age women with cancer is emerging as a challenging, but rewarding, application of assisted reproductive technologies such as in vitro fertilization. With recent advances in cryopreservation techniques, oocytes, embryos, and ovarian tissue can be banked from these patients before exposure to sterilizing chemotherapy and radiotherapy, providing future fertility options without compromising survival. PMID- 15784826 TI - Prospects for oocyte banking and in vitro maturation. AB - There is little debate about the desirability of human oocyte ("egg") banking but plenty of discussion about its prospects. Egg banking is needed by young cancer patients before they undergo potentially sterilizing treatment and is a desirable alternative to in vitro fertilization and embryo cryopreservation. However, egg banking is inefficient-oocytes are sensitive to chilling, often fail to survive freeze-thawing, and are susceptible to cytoskeletal damage and aneuploidy. Currently, even the most optimistic success rates offer patients only a slim chance of pregnancy if few oocytes are available. Ultra-rapid freezing with vitrification may offer advantages over conventional equilibrium cooling protocols and needs to be investigated further. Likewise, freezing immature oocytes followed by in vitro maturation offers practical and theoretical advantages, but this method is still inefficient. Nevertheless, all these technologies are improving, and egg banking will eventually become an option for patients seeking fertility preservation. PMID- 15784827 TI - Impact of cancer treatment on uterine function. AB - Fertility and future pregnancy potential are of concern to survivors of childhood cancer. Radiotherapy causes both ovarian and uterine damage. There are no reports of uterine damage after chemotherapy. The magnitude of risk is related to age at treatment, dose, and schedule. The dose of radiation required to destroy fifty percent of immature oocytes is <2 Gy. Reduced ovarian volume and low inhibin B and anti-Mullerian hormone concentrations in survivors with regular menses may be markers of incipient ovarian failure. Uterine damage, manifest by impaired growth and blood flow, is a likely consequence of pelvic irradiation. Uterine volume correlates with age at irradiation. Exposure of the pelvis to radiation is associated with an increased risk of miscarriage, midtrimester pregnancy loss, preterm birth, and low birthweight. The optimal dose and delivery route of estrogen replacement required to facilitate uterine growth in adolescent women treated with total body irradiation in childhood needs to be established. If female survivors of childhood cancer do achieve a pregnancy, then these pregnancies are high risk, pose a challenge for optimal mode of delivery, and require a multidisciplinary approach to management. PMID- 15784828 TI - Children exposed to chemotherapy in utero. AB - The majority of the information on the effects of in utero exposure to chemotherapy has been derived from retrospective case reports and series. Overviews of the available data have concluded that the timing of chemotherapy exposure (first trimester versus second and third trimesters) as well as the chemotherapeutic agent or agents used affect the risk of spontaneous abortion and miscarriage as well as that of congenital abnormalities. Although there are data from a prospective series of 24 pregnant breast cancer patients treated at the University of Texas M.D. Anderson Cancer Center, there are limited case series in women with hematologic malignancies, with the largest series having 89 pregnancies, that indicate that the fetuses exposed to chemotherapy in utero in the second and third trimesters can be carried to term, be born without evidence of congenital abnormalities, and develop normally. Clearly, ongoing prospective collection of data on the children born to women undergoing therapy for cancer is necessary. PMID- 15784829 TI - Pregnancy outcomes in survivors of childhood cancer. AB - During the past several decades, survival rates of many childhood cancers have risen at a remarkable pace. The ever-growing population of cancer survivors is at potential risk for developing a broad spectrum of adverse outcomes relating to cancer diagnosis and treatment, including infertility, adverse pregnancy-related outcomes, and health problems of offspring. Unfortunately, these topics have not been extensively studied, particularly among pediatric cancer survivors receiving more recent therapies. Based on the current literature, therapy for childhood cancer, in general, does not appear to have a significant impact on pregnancy outcomes and on the health of offspring of childhood cancer survivors. Additional investigations, incorporating more rigorous designs, need to be conducted to further address potential long-term risks relating to birth outcomes, including birth weight. Studies currently underway to evaluate the incidence of and risk factors for birth defects, occurrence of cancer, and other serious health-related outcomes will provide valuable information to guide researchers, clinicians, and survivors and their families. PMID- 15784830 TI - Parenthood in survivors after adulthood cancer and perinatal health in their offspring: a preliminary report. AB - Cancer survivors may fear infertility, obstetric problems, and genetic alterations in their offspring. After linkage of three registries the probability of post-treatment parenthood and the risk of obstetric and perinatal problems were estimated in cancer survivors compared to individuals without a cancer diagnosis. A total of 1531 of 13,817 patients had 2307 children after one parent's cancer diagnosis: 972 males had 1479 children and 559 females had 828. A total of 1217 patients (784 males and 433 females) became parents > or =9 months after the diagnosis (1899 births: 1221 to male cancer patients and 678 to female patients). The post-diagnosis parenthood probability was 8% at 5 years, and 14% at 10 years without further increase. Female cancer survivors gave birth to post diagnosis infants with on average 130 grams lower birth weight and 6 days shorter gestations compared with infants in the non-cancer population. Infants fathered by male cancer survivors did not differ from control infants with respect to birth weight or gestational age. There was no increase in the prevalence of major congenital malformations in the offspring of cancer survivors as compared with the offspring of the non-cancer population. Multiple births and deliveries by cesarean sections were increased. Parenthood after cancer is possible in a significant number of patients, more so for males than females. The risk of major congenital malformations was not increased relative to the non-cancer population, nor was perinatal mortality increased. However, female cancer survivors delivered more preterm births and low-birth-weight infants than what was found in the non cancer population. PMID- 15784831 TI - Reproductive issues for women with BRCA mutations. AB - Women carrying BRCA1 and BRCA2 mutations face difficult and confusing reproductive decisions that fall into three categories: issues relating to risk reducing surgeries, issues relating to use of oral contraceptives/tubal ligation, and issues relating to pregnancy and breastfeeding. Risk-reducing surgeries may confer survival benefits, but they also affect quality of life. Oral contraceptives potentially protect mutation carriers against ovarian cancer but increase the risk of early-onset breast cancer, and evidence for the efficacy of tubal ligation in reducing ovarian cancer risk in BRCA mutation carriers is contradictory. Women with BRCA mutations may increase their risk of breast cancer by becoming pregnant before age 40 years, but breastfeeding may decrease risk of breast cancer in women with BRCA mutations, regardless of age. BRCA mutation carriers desiring to become pregnant must deal with a variety of psychosocial issues, some with significant ethical implications, with minimal guidance from research. PMID- 15784832 TI - Preimplantation genetic diagnosis (PGD) for heritable neoplasia. AB - Especially applicable for heritable neoplasia, preimplantation genetic diagnosis (PGD) is possible for any Mendelian disorder whose gene has been localized, whether the molecular basis is known or not. METHODS AND RESULTS: PGD requires DNA from gametes (oocytes) or embryos before 6 days postconception, when implantation occurs. Approaches include 1) polar body biopsy, 2) blastomere biopsy (aspiration of one or two cells from the six- to eight-cell embryos at 2 or 3 days), and 3) trophectoderm biopsy, which allows recovery of 20 or more cells (20-50) from 125- to 150-cell, 5- to 6-day blastocysts. Of some 6000 PGD cycles worldwide, approximately 1500 have been performed for Mendelian indications. The approximately 25% live birth rates following PGD parallel the general U.S. experience for assisted reproductive technology. PGD has been accomplished for both cancer-specific disorders like adenomatous polyposis coli (APC), BRCA1, retinoblastoma, Li-Fraumeni syndrome, and von Hippel-Lindau syndrome (VHL), as well as disorders predisposing to neoplasia (Fanconi anemia, Wiskott-Aldrich syndrome). PGD also makes possible the identification and, hence, transfer of embryos of specific HLA genotypes. This allows cord blood harvesting for stem cell implantation into a moribund child, often an older sibling of the fetus. CONCLUSIONS: PGD is a complex, but achievable, approach especially applicable to Mendelian forms of neoplasia. PGD is an attractive addition to the prenatal diagnostic armamentarium, especially relevant to heritable neoplasia. PGD also makes possible novel indications having special relevance to heritable neoplasia. PMID- 15784833 TI - Third-party reproduction and adoption in cancer patients. AB - OBJECTIVE: To assess whether cancer patients receive timely information on adoption and third-party reproduction and whether discrimination exists in the system. METHODS: A convenience sample of 11 cancer organizations, 6 international adoption agencies, and 7 adoption specialists was informally surveyed by phone, by e-mail, and in person. RESULTS: Cancer organizations did not know whether cancer is a barrier to adoption; what a survivor needs to do to adopt; or whether or not a cancer survivor should disclose their cancer history to the adoption agency, home study worker, etc. They could not identify resources for a survivor experiencing discrimination or adoption agencies that are cancer friendly. They did not know whether a survivor should adopt domestically or internationally to increase the likelihood of success. Adoption agencies identified their chief concern as the welfare of the child and were reluctant to discuss how a cancer survivor would be viewed as a potential adoptive parent. CONCLUSIONS: Cancer patients lack immediate access to information about adoption and may face discrimination in domestic and international adoption. PMID- 15784834 TI - Defining and measuring reproductive concerns of female cancer survivors. AB - Although women diagnosed with cancer during their childbearing years are at significant risk for infertility, we know little about the relationship between infertility and long-term quality of life (QOL). To examine these relationships, we assessed psychosocial and reproductive concerns and QOL in 231 female cancer survivors. Greater reproductive concerns were significantly associated with lower QOL on numerous dimensions (P<.001). In a multiple regression model, social support, gynecologic problems, and reproductive concerns accounted for 63% of the variance in QOL scores. Women who reported wanting to conceive after cancer, but were not able to, reported significantly more reproductive concerns than those who were able to reproduce after cancer (P<.001). These preliminary data suggest that at least for vulnerable subgroups, the issue of reproductive concerns is worthy of additional investigation to assist cancer survivors living with the threat or reality of infertility. PMID- 15784835 TI - Research promoting better patient education on reproductive health after cancer. AB - Although research on cancer survivors' experiences and attitudes about infertility is relatively new, existing literature suggests that only about half of men and women of childbearing age receive the information they need from their health care providers at the time of diagnosis and treatment planning. Thus, better patient education strategies are needed. Although the ideal would be to have oncologists conduct detailed discussions of options to preserve or restore fertility with all interested patients, this standard is unrealistic. A more practical alternative is to train nonphysician providers such as nurses and social workers to address this topic. In addition, innovative strategies of direct patient education could be helpful, including use of computerized media, peer counseling, or special educational modules tailored to a particular age group or cancer site. PMID- 15784836 TI - Ensuring access to education and services on infertility for the underserved. AB - Race, culture, ethnicity, and socioeconomic status (SES) all influence how men and women cope with cancer-related infertility and whether they use infertility services. Unfortunately, these variables have not yet been studied in samples of cancer survivors. This article provides an overview of Medline-cited studies from 1980 to the present that examine the influence of ethnicity and socioeconomic status on the use of infertility services. Although underserved groups are disproportionately at risk for infertility in the United States, they are also less likely than middle- to high-SES Caucasians to seek medical treatment for this problem. Barriers to their use of infertility treatment include lack of knowledge, lack of financial resources, and cultural norms. It is very important for oncology care providers to take ethnicity and SES into account when counseling patients about infertility and to be aware of cultural and religious values with regard to assisted reproductive technology. PMID- 15784837 TI - Cancer and fertility: ethical and legal challenges. AB - Preserving the fertility of younger cancer patients requires coordinated efforts and attention to ethical issues by oncologists and fertility specialists. Although sperm is easily stored, freezing eggs or ovarian tissue is still experimental and should not be offered except as part of an experimental protocol. When gametic material is stored for later use, written directives for posthumous use may be given effect, and subsequently born children may be recognized as legal offspring of the deceased. Concerns about the welfare of offspring resulting from an expected shortened life span of the parent are not sufficient reason to deny cancer survivors assistance in reproducing. PMID- 15784838 TI - Ovarian tissue preservation and future fertility: emerging technologies and ethical considerations. AB - Both adult women and prepubertal girls facing cancer treatment may have the option to harvest and cryopreserve ovarian tissue. If they have successful therapy for their disease, the tissue may be autotransplanted back into the woman's body or follicles may be harvested and matured in vitro. These techniques all remain experimental, however, and should be performed only by specialized centers that can provide a multidisciplinary team. The procedures should be done under approval from an Institutional Review Board with proper assurance of informed consent. Although the costs for research could be assessed to patients if no research funds are available, clinical fees should not be charged at this stage in the development of these technologies. Consideration should be given to the protection of minors by ensuring parental informed consent and child assent whenever possible. Disposition of cryopreserved gonadal tissue in the event of the participant's death should be formally designated. A registry should be established to follow the health of participants and their eventual offspring. PMID- 15784839 TI - Legal issues related to parenthood after cancer. AB - Cancer and procreation raise a host of novel legal issues involving the rights of those trying to create families after cancer treatment and any resulting children, as well as the responsibilities of those who assist them. Recent court decisions, although neither consistent nor plentiful, highlight the emerging legal issues for patients, providers, and offspring. This article explores a number of legal issues related to cancer and parenthood, including: 1) patients' cryopreservation of sperm, eggs, or embryos and subsequent access to and use by them or their former partners; 2) fertility preservation in minor patients; 3) posthumous reproduction and legal parentage issues for children born from cryopreserved embryos or gametes; 4) wrongful life or wrongful birth claims of children born following their parents' cancer treatments; 5) access to, and discrimination in, medical treatment or alternative family-building options; and 6) professional responsibility and liability for providers relating to the potential fertility impact of cancer treatment. The limited, evolving court decisions, through the application of legal principles such as negligence, malpractice, discrimination, and parentage principles, provide some guidance for patients, providers, and policymakers in approaching the unique challenges presented by fertility preservation in the context of cancer treatments. PMID- 15784840 TI - A biomimetic tissue from cultured normal human urothelial cells: analysis of physiological function. AB - The urinary bladder and associated tract is lined by the urothelium. Once considered as just an impermeable epithelium, it is becoming evident that the urothelium not only functions as a volume-accommodating urinary barrier but has additional roles, including sensory signaling. Lack of access to normal human urothelium has hampered physiological investigation, and although cell culture systems have been developed, there has been a failure to demonstrate that normal human urothelial (NHU) cells grown in vitro retain the capacity to form a functional differentiated urothelium. The aim of this study was to develop a biomimetic human urothelium from NHU cell cultures. Urothelial cells isolated from normal human urothelium and serially propagated as monolayers in serum-free culture were homogeneous and adopted a proliferative, nondifferentiated phenotype. In the presence of serum and physiological concentrations of calcium, these cells could be reproducibly induced to form stratified urothelia consisting of basal, intermediate, and superficial cells, with differential expression of cytokeratins and superficial tight junctions. Functionally, the neotissues showed characteristics of native urothelium, including high transepithelial electrical resistance of >3,000 Omega.cm(2), apical membrane-restricted amiloride-sensitive sodium ion channels, basal expression of Na(+)-K(+)-ATPase, and low diffusive permeability to urea, water, and dextran. This model represents major progress in developing a biomimetic human urothelial culture model to explore molecular and functional relationships in normal and dysfunctional bladder physiology. PMID- 15784841 TI - A1 adenosine receptor knockout mice exhibit increased mortality, renal dysfunction, and hepatic injury in murine septic peritonitis. AB - Sepsis is a leading cause of multiorgan dysfunction and death in hospitalized patients. Dysregulated inflammatory processes and apoptosis contribute to the pathogenesis of sepsis-induced organ dysfunction and death. A(1) adenosine receptor (A(1)AR) activation reduces inflammation and apoptosis after ischemia reperfusion injury. Therefore, we questioned whether A(1)AR-mediated reduction of inflammation and apoptosis could improve mortality and organ dysfunction in a murine model of sepsis. A(1)AR knockout mice (A(1) knockout) and their wild-type (A(1) wild-type) littermate controls were subjected to cecal ligation and double puncture (CLP) with a 20-gauge needle. A(1) knockout mice or A(1) wild-type mice treated with 1,3-dipropyl-8-cyclopentylxanthine (a selective A(1)AR antagonist) had a significantly higher mortality rate compared with A(1) wild-type mice following CLP. Mice lacking endogenous A(1)ARs demonstrated significant elevations in plasma creatinine, alanine aminotransferase, aspartate aminotransferase, keratinocyte-derived chemokine, and tumor necrosis factor-alpha 24 h after induction of sepsis compared with wild-type mice. The renal corticomedullary junction from A(1) knockout mice also exhibited increased myeloperoxidase activity, intercellular adhesion molecule-1 protein, and mRNA encoding proinflammatory cytokines compared with renal samples from A(1) wild type littermate controls. No difference in renal tubular apoptosis was detected between A(1) knockout and A(1) wild-type mice. We conclude that endogenous A(1)AR activation confers a protective effect in mice from septic peritonitis primarily by attenuating the hyperacute inflammatory response in sepsis. PMID- 15784842 TI - An efficient system for tissue-specific overexpression of transgenes in podocytes in vivo. AB - The utility of promoter fragments isolated from the 5'-flanking region of endogenous mammalian genes to drive transgene expression in vivo is often limited by low expression levels. In this study, a bigenic system was established that allows constitutive overexpression of transgenes in a tissue-specific fashion in transgenic mice in a time- and cost-effective fashion. A modified floxed expression vector was constructed [CMVflox-enhanced green fluorescent protein (eGFP)], in which a lacZ cassette (beta-galactosidase) flanked by lox sites was placed between a CMV-promoter and the transgene of interest (eGFP). Before Cre recombination, expression of eGFP was effectively prevented by the interposed floxed lacZ cassette, whereas beta-galactosidase was strongly expressed in transiently transfected cells. Transcription of the gene of interest (eGFP) could be irreversibly activated by cotransfection with Cre recombinase. Mice transgenic for CMVflox-eGFP were generated by pronuclear injection. A rapid assay was developed to identify transgenic founders with active transgene expression by measuring transgene activity (beta-galactosidase) in tail biopsies. Transgene activity in tails correlated with transgene expression in most other tissues tested including podocytes within the kidney. To activate expression of the gene of interest in a tissue-specific fashion, founder mice were mated to the Cre mouse line 2.5P-Cre previously shown to mediate 100% Cre recombination exclusively in podocytes (Moeller MJ, Sanden SK, Soofi A, Wiggins RC, and Holzman LB. Genesis 35: 39-42, 2003). In doubly transgenic offspring, high-level eGFP expression resulting from Cre excision of the interposed lacZ cassette was detected in four of seven CMVflox-eGFP founder lines. This approach should also circumvent common limitations arising from lethality or transgene silencing as a consequence of transgene overexpression. PMID- 15784844 TI - What's for lunch at the conformational cafeteria? AB - In this issue of Molecular Pharmacology, Mukhopadhyay and Howlett present evidence for ligand-selective conformations of the CB1 cannabinoid receptor with differential coupling to G proteins. Ligand-directed signaling to different cellular effector pathways extends drug selectivity beyond that afforded by differential affinity for different receptor subtypes. The challenge for pharmacologists of the future will be not only to identify ligand-selective receptor conformations but also to develop an understanding of the relationships between those conformations, cell function, and ultimately therapeutics. As we learn more about ligand-selective receptor conformations, it should be possible to develop response-selective drugs that maximize therapeutic efficacy and minimize unwanted effects. PMID- 15784843 TI - The renal vascular response to ANG II injection is reduced in the nonclipped kidney of two-kidney, one-clip hypertension. AB - The ANG II receptor 1 (AT(1)R) level in the nonclipped kidney of two-kidney, one clip hypertension (2K1C) has shown to be unchanged despite a high circulating angiotensin (ANG) II level. To examine the vasoreactive response to ANG II in this kidney, injections of ANG II into renal artery were performed 6 wk after clipping of the kidney and compared with normotensive controls. The renal blood flow (RBF) response to 2.5 ng ANG II was measured by a Transonic transit-time flowmeter, before and after indomethacin and candesartan treatment, and analyzed by a computer program. The RBF response to 5 ng arginine-vasopressin (AVP) was examined for comparison with ANG II. The mRNA for AT(1A) and AT(1B) as well as Western blotting for AT(1)R in renal resistance vessels were determined, and plasma renin activity (PRA) was measured. Systolic blood pressure was 183 +/- 4 mmHg in 2K1C rats compared with 113 +/- 1 mmHg in controls (P < 0.001). PRA was significantly increased in 2K1C animals (P < 0.05). Injection of ANG II reduced RBF with 10 +/- 2% in the nonclipped kidney and 24 +/- 3% in controls (P < 0.001). After indomethacin, the RBF response increased from 10 +/- 2 to 20 +/- 3% (P < 0.02) in 2K1C rats and from 24 +/- 3 to 34 +/- 6% in controls (P < 0.01). The doses of candesartan needed to completely inhibit RBF response to ANG II were 30 microg/kg in the nonclipped kidney and 100 microg/kg in controls (P < 0.001). Western blotting and mRNA for AT(1A) and AT(1B) in the nonclipped kidney were similar to the controls. The results indicate that despite no difference in total AT(1)R levels, functional AT(1)R is downregulated in the nonclipped kidney of 2K1C rats. PMID- 15784845 TI - Chemical-based translational induction of luciferase expression: an efficient tool for in vivo screening of protein farnesylation inhibitors. AB - We describe the development of a cell system for in vivo screening of inhibitors of the mevalonate pathway. To this aim, we have constructed a bicistronic mRNA, transcribed from a constitutive cytomegalovirus promoter, containing the Renilla reniformis luciferase RNA open reading frame sequence as first cistron and the Firefly luciferase RNA sequence as a second cistron. The intercistronic space is made of the R17 binding sequence of the bacteriophage R17 protein. A chimeric protein able to bind to a specific sequence in the hairpin and to induce internal ribosome entry in the RNA switches on translation of the second cistron. This chimeric protein is made up of the bacteriophage RNA binding domain (R17) fused to the ribosome recruitment core of the eIF-4G1 eukaryotic translation initiation factor and to the CAAX box of H-Ras addressing the protein to the plasma membrane where it is not efficient. Internal ribosome entry upstream of the Firefly cistron is therefore under the dependence of the mevalonate pathway inhibitors. Indeed, products that are able to inhibit protein farnesylation rescue the cytoplasmic location of the R17-eIF-4G-CAAX protein, which once more becomes a translation factor for the expression of the second cistron. To exemplify the system, the present work checks the ability of various antiestrogens to interfere with the mevalonate pathway. It seems that pure antiestrogen, able to selectively bind the estrogen receptor, is unable to switch on the second Firefly cistron although selective antiestrogen-binding-site ligands are able to do so. PMID- 15784846 TI - Somatostatin increases phospholipase D activity and phosphatidylinositol 4,5 bisphosphate synthesis in clonal beta cells HIT-T15. AB - In the presence of arginine vasopressin (AVP), somatostatin increases [Ca(2+)](i), leading to a transient increase in insulin release from clonal beta cells HIT-T15 via G(i/o) and phospholipase C (PLC) pathway (Cheng et al., 2002a). The present study was to elucidate the mechanisms underlying somatostatin-induced [Ca(2+)](i) increase in the presence of AVP. We found that the effect of somatostatin was mediated by betagamma subunits but not by the alpha subunit of G(i/o). Because somatostatin alone failed to increase [Ca(2+)](i), we hypothesized that somatostatin increases phosphatidylinositol 4,5-bisphosphate (PIP(2)) synthesis, providing extra substrate for preactivated PLC-beta to generate inositol 1,4,5-trisphosphate (IP(3)). Somatostatin alone did not increase IP(3) levels, but AVP + somatostatin did. Somatostatin increased PIP(2) levels but decreased phosphatidylinositol 4-phosphate levels. We further hypothesized that PLD mediates somatostatin-induced changes in PIP(2) levels. Both the phospholipase D (PLD) inhibitors and antibody versus PLD1 antagonized AVP-somatostatin-induced increases in [Ca(2+)](i). PLD inhibitor also antagonized somatostatin-induced increase in PIP(2) levels. In addition, somatostatin increased PLD activity. These results suggest that activation of somatostatin receptors that are coupled to the betagamma dimer of G(i/o) led to PLD1 activation, thus promoting the synthesis of phosphatidic acid. Phosphatidic acid activates PIP-5 kinase, which evokes an increase in PIP(2) synthesis. The PIP(2) generated by somatostatin administration increases substrate for preactivated phospholipase C-beta, which hydrolyzes PIP(2) to form IP(3), leading to an increase in [Ca(2+)](i). The regulation of PIP(2) synthesis by G(i/o)-coupled receptors via PLD activation represents a novel signaling mechanism for somatostatin and a novel concept in the cross-talk between G(q)- and G(i/o) coupled receptors in beta cells. PMID- 15784847 TI - The neurotransmitter noradrenaline rescues septal cholinergic neurons in culture from degeneration caused by low-level oxidative stress. AB - We have developed a model system in which rat basal forebrain cholinergic neurons degenerate progressively when maintained in culture conditions that make them susceptible to low-level oxidative stress. In this study, we showed that cholinergic neurons identified by acetylcholinesterase cytochemistry or choline acetyl transferase immunocytochemistry are rescued efficiently by the neurotransmitter noradrenaline (NA). The effect of NA required neither adrenoceptor activation nor intracellular accumulation. NA operated via a mechanism that precluded activation of a cell death pathway in which reactive oxygen species (ROS) and proapoptotic caspases were crucially involved. It is noteworthy that NA remained protective even when applied late in the degenerative process but before intracellular ROS began to increase. The high efficacy of iron chelators and catalase in preventing the death of cholinergic neurons in this model suggested that NA neutralized the effects of hydroxyl radicals produced through a Fenton-type reaction. Pyrocatechol [the diphenolic moiety of NA] was sufficient in itself to prevent ROS production and cholinergic cell demise, indicating that the catechol structure was instrumental for the neuroprotective function of NA. Therefore, the noncatecholic neurotransmitter GABA failed to prevent neurodegeneration. Nerve growth factor and brain derived neurotrophic factor, two trophic peptides for septal cholinergic neurons, did not afford protection by themselves and did not improve neuroprotection provided by NA. However, in the presence of NA, they both retained their efficacy to stimulate cholinergic parameters. These data indicate that NA-based therapeutic strategies may be of interest in such neurodegenerative conditions as Alzheimer's disease, where progressive cholinergic deficits occur. PMID- 15784849 TI - A piece of my mind. Jambo, Jahi. PMID- 15784848 TI - The cover. The Lord Is My Shepherd. PMID- 15784850 TI - New vaccine should ease meningitis fears. PMID- 15784851 TI - Discovery boosts hope for heart repair. PMID- 15784852 TI - TIA time bombs remain undertreated. PMID- 15784853 TI - George Bakris, MD: researching ways to combat hypertension. Interview by Mike Mitka. PMID- 15784861 TI - Thyroid status and survival in old age. PMID- 15784862 TI - Thyroid status and survival in old age. PMID- 15784863 TI - Rhabdomyolysis and lipid-lowering drugs. PMID- 15784864 TI - Presidential candidates' commentaries. PMID- 15784865 TI - Presidential candidates' commentaries. PMID- 15784866 TI - ABO blood group and susceptibility to severe acute respiratory syndrome. PMID- 15784867 TI - Permanence of Web page references. PMID- 15784868 TI - Risk factors for relapse in health care professionals with substance use disorders. AB - CONTEXT: Substance use disorders among physicians are important and persistent problems. Considerable debate exists over whether use of major opioids, especially among anesthesiologists, is associated with a higher relapse rate compared with alcohol and nonopioids. Moreover, the risk factors for relapse with current treatment and monitoring strategies are unknown. OBJECTIVE: To test the hypothesis that chemically dependent health care professionals using a major opioid (eg, fentanyl, sufentanil, morphine, meperidine) as drug of choice are at higher risk of relapse. DESIGN, SETTING, AND PARTICIPANTS: Retrospective cohort study of 292 health care professionals enrolled in the Washington Physicians Health Program, an independent posttreatment monitoring program, followed up between January 1, 1991, and December 31, 2001. MAIN OUTCOME MEASURE: Factors associated with relapse, defined as the resumption of substance use after initial diagnosis and completion of primary treatment for chemical dependency. RESULTS: Twenty-five percent (74 of 292 individuals) had at least 1 relapse. A family history of a substance use disorder increased the risk of relapse (hazard ratio [HR], 2.29; 95% confidence interval [CI], 1.44-3.64). The use of a major opioid increased the risk of relapse significantly in the presence of a coexisting psychiatric disorder (HR, 5.79; 95% CI, 2.89-11.42) but not in the absence of a coexisting psychiatric disorder (HR, 0.85; 95% CI, 0.33-2.17). The presence of all 3 factors--major opioid use, dual diagnosis, and family history--markedly increased the risk of relapse (HR, 13.25; 95% CI, 5.22-33.59). The risk of subsequent relapses increased after the first relapse (HR, 1.69; 95% CI, 1.13 2.53). CONCLUSIONS: The risk of relapse with substance use was increased in health care professionals who used a major opioid or had a coexisting psychiatric illness or a family history of a substance use disorder. The presence of more than 1 of these risk factors and previous relapse further increased the likelihood of relapse. These observations should be considered in monitoring the recovery of health care professionals. PMID- 15784869 TI - Association of transmission intensity and age with clinical manifestations and case fatality of severe Plasmodium falciparum malaria. AB - CONTEXT: There are concerns that malaria control measures such as use of insecticide-treated bed nets, by delaying acquisition of immunity, might result in an increase in the more severe manifestations of malaria. An understanding of the relationships among the level of exposure to Plasmodium falciparum, age, and severity of malaria can provide evidence of whether this is likely. OBJECTIVE: To describe the clinical manifestations and case fatality of severe P falciparum malaria at varying altitudes resulting in varying levels of transmission. DESIGN, SETTING, AND PATIENTS: A total of 1984 patients admitted for severe malaria to 10 hospitals serving populations living at levels of transmission varying from very low (altitude >1200 m) to very high (altitude <600 m) in a defined area of northeastern Tanzania, studied prospectively from February 2002 to February 2003. Data were analyzed in a logistic regression model and adjusted for potential clustering within hospitals. MAIN OUTCOME MEASURES: Specific syndromes of severe malaria; mortality. RESULTS: The median age of patients was 1 year in high transmission, 3 years in moderate transmission, and 5 years in low transmission areas. The odds of severe malarial anemia (hemoglobin <5 g/dL) peaked at 1 year of age at high transmission and at 2 years at moderate and low transmission intensities and then decreased with increasing age (P = .002). Odds were highest in infants (0-1 year: referent; 2-4 years: odds ratio [OR], 0.83; 95% confidence interval [CI], 0.72-0.96), 5 to <15 years: OR, 0.44; 95% CI, 0.27-0.72; > or =15 years: OR, 0.44; 95% CI, 0.27-0.73; P<.001) and high transmission intensity areas (altitude <600 m: referent; 600 m to 1200 m: OR, 0.55; 95% CI, 0.35-0.84; >1200 m: OR, 0.55; 95% CI, 0.26-1.15; P for trend = .03). The odds of cerebral malaria were significantly higher in low transmission intensity areas (altitude of residence <600 m: referent; 600 m to 1200 m: OR, 3.17; 95% CI, 1.32-7.60; >1200 m: OR, 3.76; 95% CI, 1.96-7.18; P for trend = .003) and with age 5 years and older (0-1 year: referent; 2-4 years: OR, 1.57; 95% CI, 0.82-2.99; 5 to <15 years: OR, 6.07; 95% CI, 2.98-12.38; > or =15 years: OR, 6.24; 95% CI, 3.47 11.21; P<.001). The overall case-fatality rate of 7% (139 deaths) was similar at high and moderate levels of transmission but increased to 13% in low transmission areas (P = .03), an increase explained by the increase in the proportion of cases with cerebral malaria. CONCLUSIONS: Age and level of exposure independently influence the clinical presentation of severe malaria. Our study suggests that an increase in the proportion of cases with more fatal manifestations of severe malaria is likely to occur only after transmission has been reduced to low levels where the overall incidence is likely to be low. PMID- 15784870 TI - Incidence of cervical squamous intraepithelial lesions associated with HIV serostatus, CD4 cell counts, and human papillomavirus test results. AB - CONTEXT: Recent cervical cancer screening guidelines state that the interval between screenings can be safely extended to 3 years in healthy women 30 years or older who have normal cytology results and have negative test results for oncogenic human papillomavirus (HPV) DNA. OBJECTIVE: To determine the incidence of squamous intraepithelial lesions (SILs) in HIV-seropositive women with normal cytology results, by baseline HPV DNA results. DESIGN, SETTING, AND PATIENTS: Participants were HIV-seropositive (n = 855; mean age, 36 years) and HIV seronegative (n = 343; mean age, 34 years) US women with normal baseline cervical cytology who were enrolled in the Women's Interagency HIV Study (WIHS), a large, multi-institutional prospective cohort study. Since their recruitment during 1994 1995, WIHS participants have been followed up semi-annually with repeated Pap smears for a median of 7 years. MAIN OUTCOME MEASURE: The cumulative incidence of any SIL and high-grade SIL or cancer (HSIL+) was estimated according to baseline HPV DNA results, stratified by HIV serostatus and CD4 T-cell count. RESULTS: Development of any SIL in women with negative HPV results (both oncogenic and nononcogenic) at 2 years was as follows: in HIV-seropositive women with CD4 counts less than 200/microL, 9% (95% CI, 1%-18%); with CD4 counts between 200/muL and 500/microL, 9% (95% CI, 4%-13%); and with CD4 counts greater than 500/microL, 4% (95% CI, 1%-7%). The CIs for these estimates overlapped with those for HIV seronegative women with normal baseline cytology who were HPV-negative (3%; 95% CI, 1%-5%), indicating that at 2 years, there were no large absolute differences in the cumulative incidence of any SIL between groups. Furthermore, no HPV negative participants in any group developed HSIL+ lesions within 3 years. Multivariate Cox models showed that on a relative scale, the incidence of any SIL among HIV-seropositive women with CD4 counts greater than 500/microL (hazard ratio [HR], 1.2; 95% CI, 0.5-3.0), but not those with CD4 counts less than or equal to 500/microL (HR, 2.9; 95% CI, 1.2-7.1), was similar to that in HIV seronegative women. CONCLUSION: The similar low cumulative incidence of any SIL among HIV-seronegative and HIV-seropositive women with CD4 counts greater than 500/microL and who had normal cervical cytology and HPV-negative test results suggests that similar cervical cancer screening practices may be applicable to both groups, although this strategy warrants evaluation in an appropriate clinical trial. PMID- 15784871 TI - Nitrite infusions to prevent delayed cerebral vasospasm in a primate model of subarachnoid hemorrhage. AB - CONTEXT: Delayed cerebral vasospasm causes permanent neurological deficits or death in at least 15% of patients following otherwise successful treatment for ruptured intracranial aneurysm. Decreased bioavailability of nitric oxide has been associated with the development of cerebral vasospasm. OBJECTIVE: To determine whether infusions of nitrite will prevent delayed cerebral vasospasm. DESIGN, SETTING, AND SUBJECTS: A total of 14 anesthetized cynomolgus monkeys had an autologous blood clot placed around the right middle cerebral artery. Cerebral arteriography was performed before clot placement and on days 7 and 14 to assess vasospasm. The study was conducted from August 2003 to February 2004. INTERVENTIONS: A 90-mg sodium nitrite intravenous solution infused over 24 hours plus a 45-mg sodium nitrite bolus daily (n = 3); a 180-mg sodium nitrite intravenous solution infused over 24 hours (n = 3); or a control saline solution infusion (n = 8). Each was infused continuously for 14 days. MAIN OUTCOME MEASURES: Nitrite, S-nitrosothiol, and methemoglobin levels in blood and cerebrospinal fluid and degree of arteriographic vasospasm. RESULTS: In control monkeys, mean (SD) cerebrospinal fluid nitrite levels decreased from 3.1 (1.5) micromol/L to 0.4 (0.1) micromol/L at day 7 and to 0.4 (0.4) micromol/L at day 14 (P = .03). All 8 control monkeys developed significant vasospasm of the right middle cerebral artery, which was complicated by stroke and death in 1 animal. Sodium nitrite infusions increased the nitrite and methemoglobin levels (<2.1% of total hemoglobin) in the blood and cerebrospinal fluid without evoking systemic hypotension. Nitrite infusion prevented development of vasospasm (no animals developed significant vasospasm; mean [SD] reduction in right middle cerebral artery area on day 7 after subarachnoid hemorrhage of 8% [9%] in nitrite-treated monkeys vs 47% [5%] in saline-treated controls; P<.001). There was a negative correlation between the concentration of nitrite in cerebrospinal fluid and the degree of cerebral vasospasm (P<.001). Pharmacological effects of nitrite infusion were also associated with the formation of S-nitrosothiol in cerebrospinal fluid. There was no clinical or pathological evidence of nitrite toxicity. CONCLUSION: Subacute sodium nitrite infusions prevented delayed cerebral vasospasm in a primate model of subarachnoid hemorrhage. PMID- 15784872 TI - Thiopurine methyltransferase (TPMT) genotype and early treatment response to mercaptopurine in childhood acute lymphoblastic leukemia. AB - CONTEXT: Early response to multiagent chemotherapy, including mercaptopurine, as measured by minimal residual disease is an important prognostic factor for children with acute lymphoblastic leukemia (ALL). Thiopurine methyltransferase (TPMT) is involved in the metabolism of mercaptopurine and subject to genetic polymorphism, with heterozygous individuals having intermediate and homozygous mutant individuals having very low TPMT activity. OBJECTIVE: To assess the association of TPMT genotype with minimal residual disease load before and after treatment with mercaptopurine in the early treatment course of childhood ALL. DESIGN, SETTING, AND PATIENTS: TPMT genotyping of childhood ALL patients (n = 814) in Germany consecutively enrolled in the ALL-BFM (Berlin-Frankfurt-Munster) 2000 study from October 1999 to September 2002. Minimal residual disease was analyzed on treatment days 33 and 78 for risk-adapted treatment stratification. A 4-week cycle of mercaptopurine was administered between these 2 minimal residual disease measurements. Patients (n = 4) homozygous for a mutant TPMT allele, and consequently deficient in TPMT activity, were treated with reduced doses of mercaptopurine and, therefore, not included in the analyses. MAIN OUTCOME MEASURES: Minimal residual disease load before (day 33) and after (day 78) mercaptopurine treatment. Loads smaller than 10(-4) were defined as negative. RESULTS: Patients (n = 55) heterozygous for allelic variants of TPMT conferring lower enzyme activity had a significantly lower rate of minimal residual disease positivity (9.1%) compared with patients (n = 755) with homozygous wild-type alleles (22.8%) on day 78 (P = .02). This translated into a 2.9-fold reduction in risk for patients with wild-type heterozygous alleles (relative risk, 0.34; 95% confidence interval, 0.13-0.86). CONCLUSIONS: TPMT genotype has a substantial impact on minimal residual disease after administration of mercaptopurine in the early course of childhood ALL, most likely through modulation of mercaptopurine dose intensity. Our findings support a role for minimal residual disease analyses in the assessment of genotype-phenotype associations in multiagent chemotherapeutic trials. PMID- 15784873 TI - Mortality and malnutrition among populations living in South Darfur, Sudan: results of 3 surveys, September 2004. AB - CONTEXT: Mass violence against civilians in the west of Sudan has resulted in the displacement of more than 1.5 million people (25% of the population of the Darfur region). Most of these people are camped in 142 settlements. There has been increasing international concern about the health status of the displaced population. OBJECTIVE: To perform rapid epidemiological assessments of mortality and nutritional status at 3 sites in South Darfur for relief efforts. DESIGN, SETTING, AND PARTICIPANTS: In August and September 2004, mortality surveys were conducted among 137,000 internally displaced persons (IDPs) in 3 sites in South Darfur (Kass [n = 900 households], Kalma [n = 893 households], and Muhajiria [n = 900 households]). A nutritional survey was performed concomitantly among children aged 6 to 59 months using weight for height as an index of acute malnutrition (Kass [n = 894], Kalma [n = 888], and Muhajiria [n = 896]). A questionnaire detailing access to food and basic services was administered to a subset of households (n = 210 in each site). MAIN OUTCOME MEASURES: Crude and under 5-year mortality rates and nutritional status of IDPs in Kass, Kalma, and Muhajiria, South Darfur. RESULTS: Crude mortality rates, expressed as deaths per 10,000 per day, were 3.2 (95% confidence interval [CI], 2.2-4.1) in Kass, 2.0 (95% CI, 1.3 2.7) in Kalma, and 2.3 (95% CI, 1.2-3.4) in Muhajiria. Under 5-year mortality rates were 5.9 (95% CI, 3.8-8.0) in Kass, 3.5 (95% CI, 1.5-5.7) in Kalma, and 1.0 (95% CI, 0.03-1.9) in Muhajiria. During the period of displacement covered by our survey in Muhajiria, violence was reported to be responsible for 72% of deaths, mainly among young men. Diarrheal disease was reported to cause between 25% and 47% of deaths in camp residents and mainly affected the youngest and oldest age groups. Acute malnutrition was common, affecting 14.1% of the target population in Kass, 23.6% in Kalma, and 10.7% in Muhajiria. CONCLUSION: This study provides epidemiological evidence of the high rates of mortality and malnutrition among the displaced population in South Darfur and reinforces the need to mount appropriate and timely humanitarian responses. PMID- 15784874 TI - Why have academic medical centers survived? AB - Over the past decade, many observers predicted the demise of the academic medical center (AMC) due to competition from community hospitals and physicians, fragile finances, inefficiency, and organizational complexity. In 2004, we interviewed 23 AMC and community hospital administrators to determine why those predictions have proven unfounded, learn the leaders' current concerns and priorities, and to identify desirable changes. Chief concerns were reimbursement uncertainty, federal research policy, ineffective internal decision-making, and clinical quality (mentioned in more than 75% of interviews). Priorities included ensuring sufficient investment capital, revising undergraduate and graduate curricula, strengthening ties with physicians and community hospitals, attracting faculty, and meeting regulatory requirements. We advocate that the AMC: (1) modify the research model to allow greater collaboration with institutions and researchers; (2) enhance free and open export of new and proven clinical techniques and knowledge; (3) devote greater attention to meeting patients' increasing needs for counsel and guidance, not just intervention, given the plethora of complex new technologies and their promotion in the popular media; and (4) simplify their organizations. To accomplish this, it is desirable for future leaders to gain experience outside the AMC, and for faculty and institutions to be less inwardly focused and more attentive to preserving the public's trust. PMID- 15784875 TI - Diabetes and coronary revascularization. AB - CONTEXT: Patients with diabetes mellitus account for approximately 25% of the nearly 1.5 million coronary revascularization procedures performed each year in the United States and experience worse outcomes compared with nondiabetic patients. OBJECTIVES: To summarize the current state of evidence comparing the effectiveness and safety of coronary artery bypass graft (CABG) surgery and percutaneous coronary intervention (PCI) in diabetic patients and to examine developments that may affect future outcomes in this high-risk group. EVIDENCE ACQUISITION: Using the key terms diabetes mellitus, revascularization, coronary artery bypass, angioplasty, and coronary intervention, we searched MEDLINE from 1985 to 2004 for all randomized controlled trials (RCTs) comparing CABG surgery and PCI that reported outcomes in diabetic patients. Bibliographies and the Web sites of cardiology conferences were also reviewed. Studies comparing drug eluting stents and bare-metal stents were identified in a similar fashion. The literature was reviewed to identify clinical measures that may impact revascularization outcomes in diabetic patients. EVIDENCE SYNTHESIS: We identified 6 RCTs comparing CABG surgery and PCI in a total of 950 diabetic patients. A mortality benefit for CABG over balloon-only PCI has been demonstrated in diabetic patients with multivessel coronary artery disease but has not been clearly established against stent-assisted PCI or in high-risk CABG patients. Use of glycoprotein IIb/IIIa receptor inhibitors has improved survival in diabetic patients undergoing PCI. Restenosis after PCI in diabetic patients has led to substantially higher repeat revascularization rates than after CABG. The use of drug-eluting stents has led to dramatic reductions in restenosis in diabetic patients. Ongoing RCTs comparing CABG and PCI using drug-eluting stents in diabetic patients will clarify the impact of these advances on outcomes. CONCLUSIONS: There is a relative lack of data from RCTs specifically comparing CABG surgery and PCI as currently practiced in diabetic patients. The mortality advantage and decreased rates of revascularization seen with CABG in subgroups from early trials may not be applicable in the era of drug-eluting stents, glycoprotein IIb/IIIa inhibitors, and the latest medical therapies. PMID- 15784876 TI - Inhibitors of ocular neovascularization: promises and potential problems. PMID- 15784877 TI - Physician substance abuse and recovery: what does it mean for physicians--and everyone else? PMID- 15784878 TI - JAMA patient page. Malaria. PMID- 15784879 TI - Ethylene signaling pathway. AB - The structural simplicity of the plant hormone ethylene contrasts with its dramatic effects in various developmental processes, as well as in the cellular processes that ethylene initiates in response to a diversity of environmental signals. A single well-conserved signaling cascade mediates this broad spectrum of responses. Ethylene is perceived by a family of two-component histidine kinase receptors that become inactivated upon ethylene binding. In the absence of the hormone, the receptors activate CTR1, a negative regulator of ethylene responses. Sequence similarity between CTR1 and the Raf protein kinases implies involvement of a mitogen-activated protein kinase cascade in this signaling pathway. The protein EIN2 acts downstream of CTR1 and the possible kinase cascade. Although the biochemical function of EIN2 is not understood, its critical role is manifested by the complete ethylene insensitivity of EIN2 loss-of-function mutants. Downstream of EIN2, a family of plant-specific EIN3-like transcription factors mediate ethylene responses. The regulation of EIN3 stability by ethylene is accomplished by F-box-containing proteins that participate in the formation of a SKP1/cullin/F-box complex that targets proteins for degradation by the proteasome. A large number of ethylene-regulated genes have been identified, including the APETALA2 domain-containing transcription factor genes ERF1 and EDF1 to 4, which suggests the participation of a transcriptional cascade in the ethylene response. The differential regulation of some components of this complex nuclear cascade by other signaling pathways provides a possible mechanism for interaction and signal integration. As new points of intersection with other pathways and additional participants in the pathway are identified, the Connections Map will be updated to include this new information. PMID- 15784880 TI - Arabidopsis ethylene signaling pathway. AB - In plants, ethylene gas functions as a potent endogenous growth regulator. In the model system Arabidopsis thaliana, the molecular mechanisms that underlie perception and transduction of the ethylene signal to the nucleus, where the transcription of hundreds of genes is altered, are being elucidated. In the current view, ethylene is sensed by a family of five receptors that show similarity to the bacterial two-component histidine kinases, and in plants function as negative regulators of the pathway. Binding of the ethylene gas turns off the receptors, resulting in the inactivation of another negative regulator of ethylene signaling, CTR1, a Raf-like protein kinase that directly interacts with the receptors. EIN2, a protein of unknown biochemical activity that functions as a positive regulator of the pathway, acts downstream of CTR. Derepression of EIN2 by ethylene upon disabling of the receptors and CTR1 leads to the activation of EIN3 and EIN3-like transcription factors. In the absence of ethylene, the levels of EIN3 protein are extremely low because of the function of two F-box-containing proteins, EBF1 and EBF2, that target EIN3 for proteosome-mediated degradation. In the presence of ethylene, the EIN3 protein accumulates in the nucleus and initiates a transcriptional cascade, resulting in the activation and repression of hundreds of genes. To date, the only empirically demonstrated direct target of EIN3 is the APETALA2 (AP2)-domain-containing transcription factor gene ERF1. The coregulation of ERF1 by another plant hormone, jasmonic acid, illustrates how a transcriptional cascade could be utilized in a combinatorial fashion to generate a large diversity of responses using a limited number of input signals. As new components and points of intersection with other pathways are identified, the Connections Map will be updated. PMID- 15784881 TI - Teaching resources. Structure of G-protein-coupled receptors and G proteins. AB - This Teaching Resource provides lecture notes and slides for a class covering the structure and function of G protein-coupled receptors and is part of the course "Cell Signaling Systems: A Course for Graduate Students." The lecture begins with a discussion of the crystal structure of rhodopsin and G protein subunits and then proceeds to describe the molecular mechanisms of receptor activation and the subsequent release of G proteins. PMID- 15784882 TI - Identification of functional domains within the bICP0 protein encoded by bovine herpesvirus 1. AB - It is believed that the bICP0 protein encoded by bovine herpesvirus 1 (BoHV-1) stimulates productive infection by activating viral gene expression. Like the other ICP0-like proteins encoded by alphaherpesvirinae subfamily members, bICP0 contains a zinc RING finger near its amino terminus. The zinc RING finger of bICP0 activates viral transcription, stimulates productive infection, and is toxic to certain cell types. Apart from the zinc RING finger, bICP0 possesses little similarity to the herpes simplex virus type 1 ICP0 protein making it difficult to predict what regions of bICP0 are important. To begin to identify bICP0 functional domains that are not part of the zinc RING finger, a panel of transposon insertion mutants that span bICP0 was developed. A large domain spanning aa 78-256, and a separate domain that is at or near aa 457 was necessary for efficient transactivation of a simple promoter. Transposon insertion at aa 91 impaired bICP0 protein stability in transfected cells. Insertion of transposons into the acidic domain of bICP0 had little or no effect on transactivation of a simple promoter or protein expression suggesting this region does not play a major role in activating gene expression. Sequences near the C terminus (aa 607 676) contain a functional nuclear localization signal. Collectively, these studies indicated that bICP0 contains several important functional domains: (i) the zinc RING finger, (ii) two separate domains that activate transcription, and (iii) a C-terminal nuclear localization signal that is also necessary for efficient transactivation. PMID- 15784883 TI - Immunization with a bovine herpesvirus 1 glycoprotein B DNA vaccine induces cytotoxic T-lymphocyte responses in mice and cattle. AB - Virus-specific cytotoxic T lymphocytes (CTLs) are considered to be important in protection against and recovery from viral infections. In this study, several approaches to induce cytotoxicity against bovine herpesvirus 1 (BHV-1) were evaluated. Vaccination of C57BL/6 mice with BHV-1 induced a strong humoral, but no CTL, response, which may be due to downregulation of major histocompatibility complex class I molecules. In contrast, vaccinia virus expressing glycoprotein B (gB) elicited a weaker antibody response, but strong cytotoxicity, in mice. As an approach to inducing both strong humoral and cellular immune responses, a plasmid vector was then used to express gB. Both antibody and CTL responses were induced by the plasmid encoding gB in C57BL/6 and C3H mice, regardless of the type of vector backbone. This demonstrated that DNA immunization induces a broad-based immune response to BHV-1 gB. Interestingly, removal of the membrane anchor, which resulted in secretion of gB from transfected cells, did not result in reduced cytotoxicity. Here, it is shown that, compared with the cell-associated counterpart, plasmid-encoded secreted protein may induce enhanced immune responses in cattle. Therefore, calves were immunized intradermally with pMASIAtgB, a plasmid encoding the secreted form of gB (tgB), using a needle-free injection system. This demonstrated that pMASIAtgB elicited both humoral responses and activated gamma interferon-secreting CD8+ CTLs, suggesting that a DNA vaccine expressing tgB induces a CTL response in the natural host of BHV-1. PMID- 15784884 TI - Discovery of Epstein-Barr virus (EBV)-encoded RNA signal and EBV nuclear antigen leader protein DNA sequence in pet dogs. AB - The aim of this study was to investigate Epstein-Barr virus (EBV)-related virus infection in pet dogs. The presence of antibodies to EBV antigens and EBV-related DNA was determined by Western blot analysis and PCR, respectively. Among 36 pet dogs examined for serum antibodies, 32 (88.9%) were positive for EBV-specific thymidine kinase, 15 (41.7%) for EBV-encoded DNA-binding protein and 10 (27.8%) for EBV-specific DNA polymerase. A BamHI W fragment sequence encoding part of the EBV nuclear antigen leader protein was detected by PCR in corresponding leukocyte DNA samples. Among 21 dogs tested, 15 (71.4%) were positive for the BamHI W fragment sequence. The specificity of the amplified DNA fragments was confirmed by DNA sequencing. Within the amplified region of the BamHI W fragment (241 bp), DNA sequences detected in 10 dogs had 99.2% (two nucleotide variations), 99.6% (one nucleotide variation) or 100% identity to that of EBV. Furthermore, an EBV encoded RNA signal was detected by in situ hybridization in dog lymphocytes, as well as in bone-marrow sections, indicating a latent infection with EBV or an EBV like virus. In conclusion, although the sample size was small, these results showed that a widespread EBV-related gammaherpesvirus could be detected in the peripheral blood and bone marrow of pet dogs. Although no evident zoonotic transmission was detected, further studies are imperative for disclosing the biological significance of this canine EBV-like virus, which may correlate with human disorders. PMID- 15784885 TI - Development of bovine herpesvirus 4 as an expression vector using bacterial artificial chromosome cloning. AB - Several features make bovine herpesvirus 4 (BoHV-4) attractive as a backbone for use as a viral expression vector and/or as a model to study gammaherpesvirus biology. However, these developments have been impeded by the difficulty in manipulating its large genome using classical homologous recombination in eukaryotic cells. In the present study, the feasibility of exploiting bacterial artificial chromosome (BAC) cloning and prokaryotic recombination technology for production of BoHV-4 recombinants was explored. Firstly, the BoHV-4 genome was BAC cloned using two potential insertion sites. Both sites of insertion gave rise to BoHV-4 BAC clones stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. Reconstituted virus replicated comparably to wild-type parental virus and the loxP-flanked BAC cassette was excised by growing them on permissive cells stably expressing Cre recombinase. Secondly, BoHV-4 recombinants expressing Ixodes ricinus anti-complement protein I or II (IRAC I/II) were produced using a two-step mutagenesis procedure in Escherichia coli. Both recombinants induced expression of high levels of functional IRAC molecules in the supernatant of infected cells. This study demonstrates that BAC cloning and prokaryotic recombination technology are powerful tools for the development of BoHV-4 as an expression vector and for further fundamental studies of this gammaherpesvirus. PMID- 15784886 TI - Murine gammaherpesvirus-68 ORF28 encodes a non-essential virion glycoprotein. AB - Murine gammaherpesvirus-68 (MHV-68) ORF28 is a gammaherpesvirus-specific gene of unknown function. Analysis of epitope-tagged ORF28 protein indicated that it was membrane-associated and incorporated into virions in N-glycosylated, O glycosylated and unglycosylated forms. The extensive glycosylation of the small ORF28 extracellular domain--most forms of the protein appeared to be mainly carbohydrate by weight--suggested that a major function of ORF28 is to attach a variety of glycans to the virion surface. MHV-68 lacking ORF28 showed normal lytic replication in vitro and in vivo and normal latency establishment. MHV-68 ORF28 therefore encodes a small, membrane-bound and extensively glycosylated virion protein, whose function is entirely dispensable for normal, single-cycle host colonization. PMID- 15784887 TI - Analysis of the Choristoneura fumiferana nucleopolyhedrovirus genome. AB - The double-stranded DNA genome of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) was sequenced and analysed in the context of other group I nucleopolyhedroviruses (NPVs). The genome consists of 129,593 bp with a G + C content of 50.1 mol%. A total of 146 open reading frames (ORFs) of greater than 150 bp, and with no or minimal overlap were identified. In addition, five homologous regions were identified containing 7-10 repeats of a 36 bp imperfect palindromic core. Comparison with other completely sequenced baculovirus genomes revealed that 139 of the CfMNPV ORFs have homologues in at least one other baculovirus and seven ORFs are unique to CfMNPV. Of the 117 CfMNPV ORFs common to all group I NPVs, 12 are exclusive to group I NPVs. Overall, CfMNPV is most similar to Orgyia pseudotsugata MNPV based on gene content, arrangement and overall amino acid identity. Unlike other group I baculoviruses, however, CfMNPV encodes a viral enhancing factor (vef) and has two copies of p26. PMID- 15784888 TI - Gene organization and sequencing of the Choristoneura fumiferana defective nucleopolyhedrovirus genome. AB - Two distinct nucleopolyhedrovirus species of the eastern spruce budworm, Choristoneura fumiferana, exist in a symbiont-like relationship. C. fumiferana defective nucleopolyhedrovirus (CfDEFNPV) only infects C. fumiferana larvae per os in the presence of C. fumiferana nucleopolyhedrovirus Ireland strain (CfMNPV), but is infective when injected into the haemolymph. CfDEFNPV synergizes CfMNPV in per os infections and CfMNPV is always the predominant progeny. This study was undertaken to report the genomic makeup and organization of CfDEFNPV in an attempt to identify its defect and understand its synergistic role. The genome was mapped, sequenced, characterized and compared to other baculoviruses. The CfDEFNPV genome was 131,160 nt long with 149 putative open reading frames (ORFs) and a G + C content of 45.8 mol%. Homologues of all 62 conserved lepidopteran baculovirus genes were found including those implicated in per os infectivity, p74, per os infectivity factor (pif) and pif-2. Although no obvious deletions were observed to explain the defect, two ORFs, Cfdef79 and Cfdef99 (inhibitor of apoptosis-4), contained potential deletions. Cfdef50 (late expression factor 10)/Cfdef51 (vp1054) and Cfdef76/Cfdef77 (telokin-like protein) had large overlaps and a potential homologue to ac105/he65 was split. Four baculovirus repeat ORFs were present, as were two unique genes, but no enhancins were identified. CfDEFNPV contained 13 homologous regions, each with one to five palindromes. Comparison with fully sequenced baculovirus genomes identified CfDEFNPV as a group I NPV with the closest average amino acid identity to Epiphyas postvittana NPV, followed by Orgyia pseudotsugata MNPV and CfMNPV, with its closest matches being to individual Anticarsia gemmatalis MNPV gene sequences. PMID- 15784889 TI - Expression of a Toxoneuron nigriceps polydnavirus-encoded protein causes apoptosis-like programmed cell death in lepidopteran insect cells. AB - The polydnavirus Toxoneuron nigriceps bracovirus (TnBV) is an obligate symbiont associated with the braconid wasp T. nigriceps, a parasitoid of Heliothis virescens larvae. Previously, to identify polydnavirus genes that allow parasitization by altering the host immune and endocrine systems, expression patterns of TnBV genes from parasitized H. virescens larvae were analysed and cDNAs were obtained. To study the function of the protein from one such cDNA, TnBV1, overexpression of the protein was attempted by using the baculovirus Autographa californica multicapsid nucleopolyhedrovirus. Recovery of stable recombinant virus was unsuccessful, with the exception of recombinants with deletions/mutations within the TnBV1 gene. It was hypothesized that TnBV1 expression was cytotoxic to the Spodoptera frugiperda (Sf21) insect cells that were used to produce the recombinants. Therefore, the Bac-to-Bac system was used to create recombinant baculoviruses maintained in Escherichia coli expressing either TnBV1 (Ac-TnBV1) or an initiator-methionine mutant [Ac-TnBV1(ATG-)]. Microscopy revealed substantial cell death of Sf21 and High Five cells from 48 h post-infection with Ac-TnBV1, but not with the Ac-TnBV1(ATG-) recombinant virus. Ac-TnBV1-infected Sf21 cells, but not those with parental virus infection, showed an increased caspase-3-like protease activity, as well as increased terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling (TUNEL) for breaks in host genomic DNA. Although indicative of apoptosis, blebbing and apoptotic bodies were not observed in infected cells. Transiently expressing TnBV1 alone caused TUNEL staining in High Five cells. These data suggest that TnBV1 expression alone can induce apoptosis-like programmed cell death in two insect cell lines. Injection of Ac-TnBV1 budded virus, compared with parental virus, did not result in an alteration of virulence in H. virescens larvae. PMID- 15784891 TI - A new intertype recombinant between genotypes C and D of hepatitis B virus identified in China. AB - Hepatitis B virus (HBV) genotypes have a characteristic geographical distribution. More than 90% of chronic HBV patients in China are infected with genotypes B or C. Here, eight HBV isolates that were initially classified as genotype D by PCR-restriction fragment length polymorphism analysis were analysed in detail. The complete HBV genome was sequenced and compared with 32 sequences retrieved from GenBank, representing HBV genotypes A-G. Phylogenetic analysis of the S gene (nt 10-800) classified all eight isolates as genotype D. However, phylogenetic analyses of nt 800-10 and the open reading frames (ORFs) of the precore/core and X genes classified all eight isolates as genotype C. This discordance between phylogenetic trees reconstructed on different ORFs suggested that intertype recombination has occurred in all eight isolates. By using the simplot program, the site of recombination with genotype D was located in the preS2/S region, spanning nt 10-799 in seven of eight isolates and nt 10-1499 in the other isolate. These results demonstrate that intertype recombination should be considered as a type of variation that increases the genetic diversity of HBV. Hybrids of different HBV genotypes might exhibit specific virological properties and their significance in the diagnosis and management of chronic hepatitis B deserves further investigation. PMID- 15784890 TI - Cloning, characterization and analysis by RNA interference of various genes of the Chelonus inanitus polydnavirus. AB - Successful parasitism of some endoparasitic wasps depends on an obligately symbiotic association with polydnaviruses. These unique viruses have a segmented genome consisting of circles of double-stranded (ds) DNA and do not replicate in the parasitized host. They are produced in the wasp's ovary and injected into the host along with the egg. Chelonus inanitus is an egg-larval parasitoid; its polydnavirus (CiV) has been shown to protect the parasitoid larva from the host's immune system and to induce developmental arrest in the prepupal stage. The genome of CiV consists of at least 10-12 segments and five have been sequenced up to now. Here, the complete (CiV12g2) or partial (CiV12g1, CiV16.8g1) cloning of three new CiV genes is reported. All three occur only on one viral segment and have no similarity to other known polydnavirus genes, with the exception of a high similarity of CiV12g1 to CiV14g1 and CiV12g2 to CiV14g2. Furthermore, the first attempt of in vivo application of RNA interference to study the function of polydnavirus genes is shown. Injection of dsRNA of two late- and one early- and late-expressed CiV genes into CiV/venom-containing host eggs partially rescued last-instar larvae from developmental arrest. Injection of the same dsRNAs into parasitized eggs partially reduced parasitoid survival, mainly by preventing the successful emergence of the parasitoid from the host. These viral genes thus seem to be involved in inducing developmental arrest and in keeping the cuticle soft, which appears to be necessary for parasitoid emergence and host feeding. PMID- 15784892 TI - An investigation of the therapeutic value of vaccinia-immune IgG in a mouse pneumonia model. AB - Vaccinia-immune globulin (VIG) was used to treat severe complications of smallpox vaccination, but its use was controversial because it resolved disease in only some clinical cases. VIG is a pool of hyperimmune sera collected from individuals with a high neutralizing titre against the intracellular mature form (IMV) of vaccinia virus (VACV), but activity against the extracellular enveloped form (EEV) was often not considered. Here, the efficacy of anti-VACV antibodies (Abs) in protecting mice from intranasal infection with the VACV strain Western Reserve (WR) was evaluated. Mice were immunized passively with hyperimmune rabbit Abs (IgG) generated against inactivated IMV or produced following infection by VACV; subsequently, animals were challenged with VACV WR. The results demonstrated that: (i) good protection requires Abs to EEV in addition to IMV; (ii) Abs were effective when given before or up to 4 days after infection; and (iii) protection of mice from VACV WR correlated with a reduction of virus replication in lungs, but not in brain. In agreement with studies conducted before smallpox was eradicated and recent studies using EEV antigens for immunization, this study reiterates the importance of anti-EEV Abs in protecting against orthopoxvirus infection and illustrates the need to evaluate both anti-IMV and anti-EEV neutralizing Abs in VIG. PMID- 15784893 TI - Binding of human papillomavirus 16 E6 to p53 and E6AP is impaired by monoclonal antibodies directed against the second zinc-binding domain of E6. AB - The E6 protein of cancer-associated human papillomavirus type 16 (16E6) binds to p53 and, in association with E6AP, promotes its degradation through the ubiquitin proteasome pathway. The aim of this work was to develop monoclonal antibodies against 16E6 and to test their effect on the binding of 16E6 to p53 and E6AP, and on the degradation of p53. It was shown that an antibody directed against the N terminus of 16E6 inhibited E6AP-dependent binding to p53 and degradation of p53, whereas two different antibodies directed to the second zinc-binding domain of 16E6 reduced 16E6 E6AP-independent binding to p53 and binding to E6AP but not degradation of p53. PMID- 15784894 TI - Minute virus of mice small non-structural protein NS2 localizes within, but is not required for the formation of, Smn-associated autonomous parvovirus associated replication bodies. AB - The non-structural proteins NS1 and NS2 of the parvovirus minute virus of mice (MVM) are required for efficient virus replication. It has previously been shown that NS1 and NS2 interact and colocalize with the survival motor neuron (Smn) gene product in novel nuclear structures that are formed late in infection, termed Smn-associated APAR (autonomous parvovirus-associated replication) bodies (SAABs). It is not clear what molecular viral intermediate(s) contribute to SAAB formation. The current results address the role of NS2 in SAAB formation. In highly synchronized wild-type MVM infection of murine A9(2L) cells, NS2 colocalizes with Smn and other SAAB constituents. An MVM mutant that does not produce NS2 still generates SAABS, albeit with a temporal delay. The lag in SAAB formation seen in the absence of NS2 is probably related to the temporal delay in virus replication, suggesting that, whilst NS2 is required for efficient viral infection, it is dispensable for SAAB formation. PMID- 15784895 TI - Hepatitis C virus (HCV) NS5A protein downregulates HCV IRES-dependent translation. AB - Translation of the hepatitis C virus (HCV) polyprotein is mediated by an internal ribosome entry site (IRES) that is located mainly within the 5' non-translated region of the viral genome. In this study, the effect of the HCV non-structural 5A (NS5A) protein on the HCV IRES-dependent translation was investigated by using a transient transfection system. Three different cell lines (HepG2, WRL-68 and BHK-21) were co-transfected with a plasmid vector containing a bicistronic transcript carrying the chloramphenicol acetyltransferase (CAT) and the firefly luciferase genes separated by the HCV IRES sequences, and an expression vector producing the NS5A protein. Here, it was shown that the HCV NS5A protein inhibited HCV IRES-dependent translation in a dose-dependent manner. In contrast, NS5A had no detectable effect on cap-dependent translation of the upstream gene (CAT) nor on translation from another viral IRES. Further analysis using deleted forms of the NS5A protein revealed that a region of about 120 aa located just upstream of the nuclear localization signal of the protein is critical for this suppression. Overall, these results suggest that HCV NS5A protein negatively modulates the HCV IRES activity in a specific manner. PMID- 15784896 TI - Perturbation of epidermal growth factor receptor complex formation and Ras signalling in cells harbouring the hepatitis C virus subgenomic replicon. AB - Hepatitis C virus non-structural NS5A protein inhibits epidermal growth factor (EGF)-stimulated activation of the Ras-ERK mitogen-activated protein kinase pathway at a point upstream of Ras activation. To determine the mechanism of this inhibition, the events occurring between the EGF receptor and Ras in Huh-7 cells harbouring the HCV subgenomic replicon were investigated. It was shown that, following EGF stimulation, these cells exhibited decreased EGF receptor tyrosine phosphorylation, aberrant recruitment of the adaptor proteins ShcA and Grb2 to the EGF receptor, reduced phosphorylation of ShcA and reduced Ras activation in comparison with control cells. These data are consistent with effects of NS5A and/or other components of the replicon on multiple events occurring upstream of Ras. PMID- 15784897 TI - Further studies on hepatitis C virus NS5A-SH3 domain interactions: identification of residues critical for binding and implications for viral RNA replication and modulation of cell signalling. AB - The NS5A protein of hepatitis C virus has been shown to interact with a subset of Src homology 3 (SH3) domain-containing proteins. The molecular mechanisms underlying these observations have not been fully characterized, therefore a previous analysis of NS5A-SH3 domain interactions was extended. By using a semi quantitative ELISA assay, a hierarchy of binding between various SH3 domains for NS5A was demonstrated. Molecular modelling of a polyproline motif within NS5A (termed PP2.2) bound to the FynSH3 domain predicted that the specificity determining RT-loop region within the SH3 domain did not interact directly with the PP2.2 motif. However, it was demonstrated that the RT loop did contribute to the specificity of binding, implicating the involvement of other intermolecular contacts between NS5A and SH3 domains. The modelling analysis also predicted a critical role for a conserved arginine located at the C terminus of the PP2.2 motif; this was confirmed experimentally. Finally, it was demonstrated that, in comparison with wild-type replicon cells, inhibition of the transcription factor AP-1, a function previously assigned to NS5A, was not observed in cells harbouring a subgenomic replicon containing a mutation within the PP2.2 motif. However, the ability of the mutated replicon to establish itself within Huh-7 cells was unaffected. The highly conserved nature of the PP2.2 motif within NS5A suggests that functions involving this motif are of importance, but are unlikely to play a role in replication of the viral RNA genome. It is more likely that they play a role in altering the cellular environment to favour viral persistence. PMID- 15784898 TI - Heterologous gene expression by infectious and replicon vectors derived from tick borne encephalitis virus and direct comparison of this flavivirus system with an alphavirus replicon. AB - The flavivirus tick-borne encephaltis virus (TBEV) was established as a vector system for heterologous gene expression. The variable region of the genomic 3' non-coding region was replaced by an expression cassette consisting of the reporter gene enhanced green fluorescent protein (EGFP) under the translational control of an internal ribosomal entry site element, both in the context of an infectious virus genome and of a replicon lacking the genes of the surface proteins prM/M and E. The expression level and the stability of expression were measured by fluorescence-activated cell-sorting analysis and compared to an established alphavirus replicon vector derived from Venezuelan equine encephaltis virus (VEEV), expressing EGFP under the control of its natural subgenomic promoter. On the first day, the alphavirus replicon exhibited an approximately 180-fold higher expression level than the flavivirus replicon, but this difference decreased to about 20- and 10-fold on days 2 and 3, respectively. Four to six days post-transfection, foreign gene expression by the VEEV replicon vanished almost completely, due to extensive cell killing. In contrast, in the case of the TBEV replicon, the percentage of positive cells and the amount of EGFP expression exhibited only a moderate decline over a time period of almost 4 weeks. The infectious TBEV vector expressed less EGFP than the TBEV replicon at all times. Significant expression from the infectious vector was maintained for four cell-culture passages. The results indicate that the VEEV vector is superior with respect to achieving high expression levels, but the TBEV system may be advantageous for applications that require a moderate, but more enduring, gene expression. PMID- 15784899 TI - Dengue virus-induced apoptosis in hepatic cells is partly mediated by Apo2 ligand/tumour necrosis factor-related apoptosis-inducing ligand. AB - Although hepatic injury is reported in cases with dengue haemorrhagic fever and dengue shock syndrome, its mechanism remains poorly understood. Several findings suggest that dengue virus (DEN) induces apoptosis of hepatocytes in vivo. In this work, DEN type 2 (DEN-2) strain NGC was shown to induce apoptosis in the hepatic cell line HepG2, and infection of HepG2 cells was found to induce Apo2 ligand (Apo2L, also known as tumour necrosis factor-related apoptosis-inducing ligand or TRAIL) expression. Furthermore, Apo2L/TRAIL induced apoptosis in HepG2 cells, which expressed the Apo2L/TRAIL receptor DR5/TRAIL-R2 on their surface. Analysis of the Apo2L/TRAIL promoter revealed that this gene was activated by DEN-2 infection, whose responsive element was overlapping NF-kappaB- and Sp1-binding sites located at nt -75 to -65. The proteasome inhibitor N-acetyl-L-leucinyl-L leucinyl-L-norleucinal (LLnL) inhibited Apo2L/TRAIL mRNA expression, and LLnL and anti-Apo2L/TRAIL antibody inhibited DEN-2-induced apoptosis. It was proposed that DEN infection promotes apoptosis partly through the induction of Apo2L/TRAIL expression. PMID- 15784901 TI - 'Rescue' of mini-genomic constructs and viruses by combinations of morbillivirus N, P and L proteins. AB - Chloramphenicol acetyltransferase (CAT)-expressing negative-sense mini-genomic constructs of measles virus (MV) and rinderpest virus (RPV) were rescued by standard technology with helper plasmids expressing the nucleocapsid (N), phospho (P) and large (L) proteins of MV, canine distemper virus (CDV) or RPV in order to determine whether the proteins of different viruses can function together. Homogeneous sets consisting of N, P and L plasmids derived from one virus were able to generate reporter gene expression from either mini-genomic construct. Heterogeneous sets of proteins from different viruses were not functional, with the exception that a low level of activity was obtained when MV N and P protein were combined with RPV L protein in the rescue of the MV mini-genomic construct, or CDV N was combined with RPV P and L in the rescue of the RPV mini-genome. However, only homogeneous sets of plasmids were able to rescue infectious virus from full-length anti-genome-expressing plasmids. PMID- 15784902 TI - A role for virus promoters in determining the pathogenesis of Rinderpest virus in cattle. AB - Rinderpest virus (RPV) is a morbillivirus that causes cattle plague, a disease of large ruminants. The viral genome is flanked at the 3' and 5' genome termini by the genome promoter (GP) and antigenome promoter (AGP), respectively. These promoters play essential roles in directing replication and transcription as well as RNA encapsidation and packaging. It has previously been shown that individual changes to the GP of RPV greatly affect promoter activity in a minigenome assay and it was therefore proposed that individual nucleotide changes in the GP and AGP might also have significant effects on the ability of the virus to replicate and cause disease in cattle. The Plowright vaccine strain of RPV has been derived by tissue-culture passage from the virulent Kabete 'O' isolate (KO) and is highly attenuated for all ruminant species in which it has been used. Here, it was shown that swapping the GP and the first 76 nt of the AGP between virulent and avirulent strains affected disease progression. In particular, it was shown that flanking the virulent strain with the vaccine GP and AGP sequences, while not appreciably affecting virus growth in vitro, led to attenuation in vivo. The reverse was not true, since the KO promoters did not alter the vaccine's attenuated nature. The GP/AGP therefore play a role in attenuation, but are not the only determinants of attenuation in this vaccine. PMID- 15784903 TI - The Plowright vaccine strain of Rinderpest virus has attenuating mutations in most genes. AB - The currently used vaccine strain of Rinderpest virus was derived by serial passage of the highly virulent Kabete 'O' strain (KO). A full-length cDNA copy of the KO strain was made from which a virus identical in pathogenicity to the wild type virus was rescued. A series of chimeric viruses was prepared in which the coding sequences for the N, P, F, H or L proteins were replaced with the corresponding sequences from the vaccine strain. The KO-based virus with the vaccine strain H gene and that with the carboxy-terminal half of the L gene replaced with the corresponding sequence from the vaccine strain retained all or almost all of the virulence of the original KO virus. Animals infected with the KO-based virus containing the vaccine strain N, P or F gene, or the amino terminal half of the L gene, developed high and prolonged pyrexia and leukopenia, but with reduced or absent lesions and other clinical signs; although partially attenuated, none was nearly as attenuated as the vaccine strain itself. These data indicate that the high attenuation and stability of the current vaccine are due to the accumulation of a number of separate mutations, none of which is itself so sufficiently debilitating that there is strong selective pressure in favour of the revertant. PMID- 15784904 TI - Epitope mapping of human respiratory syncytial virus 22K transcription antitermination factor: role of N-terminal sequences in protein folding. AB - The reactivity of a panel of 12 monoclonal antibodies raised against the human respiratory syncytial virus 22 kDa (22K) protein was tested by Western blotting with a set of 22K deletion mutants. The results obtained identified sequences in the C-terminal half of the 22K polypeptide required for integrity of most antibody epitopes, except for epitope 112, which was lost in mutants with short N terminal deletions. This antibody, in contrast to the others, failed to immunoprecipitate the native 22K protein, indicating that the N terminus of this protein is buried in the native molecule and exposed only under the denaturing conditions of Western blotting. In addition, N-terminal deletions that abolished reactivity with monoclonal antibody 112 also inhibited phosphorylation of the 22K protein previously identified at Ser-58 and Ser-61, suggesting that the N terminus is important in regulating the 22K protein phosphorylation status, most likely as a result of its requirement for protein folding. PMID- 15784905 TI - Determination of phosphorylated residues from human respiratory syncytial virus P protein that are dynamically dephosphorylated by cellular phosphatases: a possible role for serine 54. AB - The 241 aa human respiratory synctyial virus (HRSV) Long strain P protein is phosphorylated at serines 116, 117 and/or 119, and 232. Phosphates added to these residues have slow turnover and can be detected in the absence of protein phosphatase inhibition. Inhibition of phosphatases PP1 and PP2A increases the level of phosphorylation at serines 116, 117 and/or 119, suggesting a more rapid turnover for phosphates added to these residues compared to that of S232. High turnover phosphorylation is detected in the P-protein NH2-terminal region, mainly at S54 and, to a lesser extent, at S39, in the Long strain. When the P protein bears the T46I substitution (in the remaining HRSV strains), phosphates are added to S30, S39, S45 and S54. Phosphatase PP1 removes phosphate at residues in the central part of the P-protein molecule, whereas those in the NH2-terminal region are removed by phosphatase PP2A. The significance of the phosphorylation of the NH2-terminal region residues for some P-protein functions was studied. The results indicated that this modification is not essential for P-protein oligomerization or for its role in viral RNA synthesis. Nonetheless, dephosphorylation at S54 could facilitate P-M protein interactions that probably occur during the egress of viral particles. PMID- 15784906 TI - Pathogenesis of Hong Kong H5N1 influenza virus NS gene reassortants in mice: the role of cytokines and B- and T-cell responses. AB - The severity of disease caused in humans by H5N1 influenza viruses remains unexplained. The NS gene of Hong Kong H5N1/97 viruses was shown to contribute to high pathogenicity of reassortants in a pig model. However, the molecular pathogenesis and host immune response underlying this phenomenon remain unclear. Here, in a mouse model, H1N1 A/Puerto Rico/8/34 (PR/8) reassortants that contained the H5N1/97 NS gene, the H5N1/01 NS gene, or an altered H5N1/97 NS gene encoding a Glu92-->Asp substitution in NS1 was studied. The pathogenicity of reassortant viruses, the induction of cytokines and chemokine CXCL1 (KC) in the lungs and specific B- and T-cell responses was characterized. In mice infected with reassortant virus containing the H5N1/97 NS gene, the mouse lethal dose (50%) and lung virus titres were similar to those of PR/8, which is highly pathogenic to mice. This reassortant virus required two more days than PR/8 to be cleared from the lungs of infected mice. Reassortants containing the altered H5N1/97 NS gene or the H5N1/01 NS gene demonstrated attenuated pathogenicity and lower lung titres in mice. Specific B- and T-cell responses were consistent with viral pathogenicity and did not explain the delayed clearance of the H5N1/97 NS reassortant. The reassortant induced elevated pulmonary concentrations of the inflammatory cytokines IL1alpha, IL1beta, IL6, IFN-gamma and chemokine KC, and decreased concentrations of the anti-inflammatory cytokine IL10. This cytokine imbalance is reminiscent of the clinical findings in two humans who died of H5N1/97 infection and may explain the unusual severity of the disease. PMID- 15784907 TI - In vitro demonstration of neural transmission of avian influenza A virus. AB - Neural involvement following infections of influenza viruses can be serious. The neural transport of influenza viruses from the periphery to the central nervous system has been indicated by using mouse models. However, no direct evidence for neuronal infection has been obtained in vitro and the mechanisms of neural transmission of influenza viruses have not been reported. In this study, the transneural transmission of a neurotropic influenza A virus was examined using compartmentalized cultures of neurons from mouse dorsal root ganglia, and the results were compared with those obtained using the pseudorabies virus, a virus with well-established neurotransmission. Both viruses reached the cell bodies of the neurons via the axons. This is the first report on axonal transport of influenza A virus in vitro. In addition, the role of the cytoskeleton (microtubules, microfilaments and intermediate filaments) in the neural transmission of influenza virus was investigated by conducting cytoskeletal perturbation experiments. The results indicated that the transport of avian influenza A virus in the neurons was independent of microtubule integrity but was dependent on the integrity of intermediate filaments, whereas pseudorabies virus needed both for neural spread. PMID- 15784900 TI - Dynamics of hepatitis C virus NS5A quasispecies during interferon and ribavirin therapy in responder and non-responder patients with genotype 1b chronic hepatitis C. AB - The quasispecies nature of hepatitis C virus (HCV) may have important implications concerning resistance to antiviral agents. To determine whether HCV NS5A quasispecies composition and dynamics are related to responsiveness to combined interferon (IFN) and ribavirin therapy, extensive sequence analyses of cloned RT-PCR amplification products of HCV-1b NS5A quasispecies of sequential isolates from 15 treated (nine sustained responders and six non-responders) and three untreated patients were performed. Accumulation of mutations in NS5A during therapy was relatively frequent in the V3 domain, but unusual elsewhere. Amino acid changes were the result of the imposition of minor variants that were already present before treatment and always occurred within the first week of therapy. Before treatment, the complexity and diversity of quasispecies were lower in isolates from responders than in those from non-responders, particularly in the V3 domain, where differences in nucleotide entropy (0.35 vs 0.64, P=0.003), genetic distance (0.0145 vs 0.0302, P=0.05) and non-synonymous substitutions (0.0102 vs 0.0203, P=0.036) were statistically significant. These differences became more apparent during treatment, because complexity and diversity remained stable or tended to increase in non-responders, whereas they tended to decrease in responders. These observations suggest that the composition and dynamics of HCV NS5A quasispecies, particularly in the V3 domain, may play a role in the response to combined IFN/ribavirin therapy. PMID- 15784908 TI - Identification and functional analysis of VP3, the guanylyltransferase of Banna virus (genus Seadornavirus, family Reoviridae). AB - Banna virus (BAV) particles contain seven structural proteins: VP4 and VP9 form an outer-capsid layer, whilst the virus core contains three major proteins (VP2, VP8 and VP10) and two minor proteins (VP1 and VP3). Sequence analysis showed that VP3 contains motifs [Kx(I/V/L)S] and (Hx(n)H) that have previously been identified in the guanylyltransferases of other reoviruses. Incubation of purified BAV-Ch core particles with [alpha-32P]GTP resulted in exclusive covalent labelling of VP3, demonstrating autoguanylation activity (which is considered indicative of guanylyltransferase activity). Recombinant VP3 prepared in a cell free expression system was also guanylated under similar reaction conditions, and products were synthesized (in the presence of non-radiolabelled GDP) that co migrated with GMP, GDP and GpppG during TLC. This reaction, which required magnesium ions for optimum activity, demonstrates that VP3 possesses nucleoside triphosphatase (GTPase) activity and is the BAV guanylyltransferase (RNA 'capping' enzyme). PMID- 15784909 TI - Structural organization of an encephalitic human isolate of Banna virus (genus Seadornavirus, family Reoviridae). AB - Banna virus (BAV) is the type species of the genus Seadornavirus within the family Reoviridae. The Chinese BAV isolate (BAV-Ch), which causes encephalitis in humans, was shown to have a structural organization and particle morphology reminiscent of that of rotaviruses, with fibre proteins projecting from the surface of the particle. Intact BAV-Ch virus particles contain seven structural proteins, two of which (VP4 and VP9) form the outer coat. The inner (core) particles contain five additional proteins (VP1, VP2, VP3, VP8 and VP10) and are 'non-turreted', with a relatively smooth surface appearance. VP2 is the 'T = 2' protein that forms the innermost 'subcore' layer, whilst VP8 is the 'T = 13' protein forming the core-surface layer. Sequence comparisons indicate that BAV VP9 and VP10 are equivalent to the VP8* and VP5* domains, respectively, of rotavirus outer-coat protein VP4 (GenBank accession no. P12976). VP9 has also been shown to be responsible for virus attachment to the host-cell surface and may be involved in internalization. These similarities reveal a previously unreported genetic link between the genera Rotavirus and Seadornavirus, although the expression of BAV VP9 and VP10 from two separate genome segments, rather than by the proteolytic cleavage of a single gene product (as seen in rotavirus VP4), suggests a significant evolutionary jump between the members of these two genera. PMID- 15784910 TI - Characterization of the nucleic acid-binding activity of the avian reovirus non structural protein sigma NS. AB - The avian reovirus non-structural protein sigma NS has previously been shown to bind single-stranded (ss) RNA in vitro in a sequence-independent manner. The results of the present study further reveal that sigma NS binds poly(A), poly(U) and ssDNA, but not poly(C), poly(G) or duplex nucleic acids, suggesting that sigma NS has some nucleotide-sequence specificity for ssRNA binding. The current findings also show that sigma NS is present in large ribonucleoprotein complexes in the cytoplasm of avian reovirus-infected cells, indicating that it exists in intimate association with ssRNAs in vivo. Removal of RNA from the complexes generates a sigma NS protein form that sediments between 4.5 and 7 S, suggesting that RNA-free sigma NS associates into small oligomers. Expression and purification of recombinant sigma NS in insect cells allowed us to generate specific antibodies and to perform a variety of assays. The results of these assays revealed that: (i) RNA-free sigma NS exists as homodimers and homotrimers; (ii) the minimum RNA size for sigma NS binding is between 10 and 20 nt; (iii) sigma NS does not have a preference for viral mRNA sequences; and (iv) its RNA binding activity is conformation-dependent. Baculovirus expression of point and deletion sigma NS mutants in insect cells showed that the five conserved basic amino acids that are important for RNA binding and ribonucleoprotein-complex formation are dispersed throughout the entire sigma NS sequence, suggesting that this protein binds ssRNA through conformational domains. Finally, the properties of the avian reovirus protein sigma NS are compared with those of its mammalian reovirus counterpart. PMID- 15784911 TI - Differential effects of R5 and X4 human immunodeficiency virus type 1 infection on CD4+ cell proliferation and activation. AB - Human immunodeficiency virus type 1 (HIV-1) isolates can be distinguished by their chemokine coreceptor usage. Non-syncytium-inducing (NSI), macrophage-tropic viruses utilize CCR5 and are called R5 viruses; syncytium-inducing (SI) isolates use CXCR4 and are known as X4 viruses. R5 and X4 HIV isolates are both transmitted but, in most cases, R5 viruses predominate in the blood prior to the development of AIDS-related pathogenesis. The reason for the selective growth of the R5 strain is not known, but could reflect a replication advantage of R5 viruses over X4 viruses in CD4+ cells. To explore this possibility, eight phenotypically distinct viruses were used to infect CD4+ cells and cellular proliferation and activation were evaluated. In unstimulated CD4+ cells, R5 virus isolates increased the level of cell activation compared with X4 virus isolates and uninfected control cells. In CD4+ cells that were stimulated with interleukin 2, both R5 and X4 viruses were found to decrease the level of cell proliferation and reduce the majority of the activation markers studied when compared with uninfected control CD4+ cells from the same donors. However, although equal amounts of CD4+ cells were infected, R5 virus-infected CD4+ cells showed a two- to fourfold increase in cellular proliferation over X4 viruses, as measured by [3H]thymidine incorporation (P=0.001) and nuclear expression of Ki67 (P=0.001). In addition, a larger proportion of CD4+ T cells infected with R5 viruses had significantly higher levels of activation-marker expression (e.g. CD25, CD71 and HLA-DR) than CD4+ T lymphocytes infected with X4 viruses (P<0.02). Taken together, these results indicate that CD4+ cells infected with R5 virus isolates may have a selective advantage over X4 virus-infected CD4+ T cells for survival and, hence, virus spread. PMID- 15784912 TI - Inhibition of Marburg virus protein expression and viral release by RNA interference. AB - High mortality rates and lack of an available vaccine against Marburg haemorrhagic fever (MHF) highlight the need for a defensive therapy against MHF and greater knowledge of the causative agent, the Marburg virus (MARV). Here, RNA interference (RNAi) is employed to destroy MARV transcripts, disrupting replication and allowing analysis of various roles of MARV proteins. Small interfering RNAs (siRNAs) homologous to three MARV transcripts (NP, VP35 and VP30) were co-transfected into cells with plasmids encoding the corresponding nucleocapsid proteins. The resulting decrease in MARV nucleocapsid-protein levels was shown to be specific, as siRNA that was not homologous to the MARV genome did not decrease the levels of viral nucleocapsid proteins. Additionally, transcript levels of double-stranded RNA (dsRNA)-sensor proteins, the dsRNA-activated protein kinase and 2',5'-oligoadenylate synthetase 1 remained unchanged, suggesting that the decrease in viral proteins was not a result of activation of the antiviral properties of the interferon system. Subsequently, siRNAs were shown to reduce intracellular viral proteins in MARV-infected cells and viral material released into the medium. Targeted reduction of VP30 downregulated the intracellular levels of all other viral proteins, suggesting that VP30 plays an essential role for transcription/replication. The efficient reduction of MARV replication also suggests that RNAi may provide an agent against MHF. PMID- 15784913 TI - Replication of a recombinant hepatitis E virus genome tagged with reporter genes and generation of a short-term cell line producing viral RNA and proteins. AB - Hepatitis E virus (HEV) replication has been demonstrated in HepG2 cells transfected with full-length in vitro transcripts of an infectious cDNA clone. This cDNA clone was modified to generate several subgenomic HEV replicons with fused reporter genes. In vitro-transcribed capped RNAs generated from these were transfected into HepG2 cells. Negative-strand RNA was detected, indicating the occurrence of replication. The replicon containing an in-frame fusion of HEV ORF2 with enhanced green fluorescent protein (EGFP) was positive for fluorescence, whereas no signal was observed when the replicase domain was deleted. An HEV ORF3 EGFP in-frame fusion did not yield fluorescence. Deletions introduced into ORF2 did not affect the replication competency of the viral RNA. To explore the possibility of using a reporter-gene assay to monitor the synthesis of plus- and minus-strand RNA, the EGFP gene fused to the encephalomyocarditis virus internal ribosome entry site (IRES) was inserted into partially deleted ORF2 of HEV, in both the sense [HEV-IRES-EGFP(+)] and antisense [HEV-IRES-EGFP(-)] orientations. HepG2 cells transfected with HEV-IRES-EGFP(+) and HEV-IRES-EGFP(-) vectors were positive for EGFP fluorescence. To quantify HEV replication, EGFP was replaced with Renilla luciferase (RLuc). HEV-IRES-RLuc(+) showed approximately 10-fold higher luminescence than HEV-IRES-RLuc(-). There was complete loss of activity when the helicase-replicase domain in HEV-IRES-RLuc(-) was deleted. A short-term HepG2 cell line containing the full-length viral genome in the pcDNA3 vector was established. Viral RNA and proteins (RdRp, pORF2 and pORF3) could be detected in the geneticin-resistant cells, even after the seventh passage. In the absence of a reliable cell-culture system to study HEV biology, these reporter replicons, as well as the cell line, bestow immense utility. PMID- 15784914 TI - Natural isolates of Brome mosaic virus with the ability to move from cell to cell independently of coat protein. AB - Brome mosaic virus (BMV) requires encapsidation-competent coat protein (CP) for cell-to-cell movement and the 3a movement protein (MP) is involved in determining the CP requirement for BMV movement. However, these conclusions have been drawn by using BMV strain M1 (BMV-M1) and a related strain. Here, the ability of the MPs of five other natural BMV strains to mediate the movement of BMV-M1 in the absence of CP was tested. The MP of BMV M2 strain (BMV-M2) efficiently mediated the movement of CP-deficient BMV-M1 and the MPs of two other strains functioned similarly to some extent. Furthermore, BMV-M2 itself moved between cells independently of CP, demonstrating that BMV-M1 and -M2 use different movement modes. Reassortment between CP-deficient BMV-M1 and -M2 showed the involvement of RNA3 in determining the CP requirement for cell-to-cell movement and the involvement of RNAs 1 and 2 in movement efficiency and symptom induction in the absence of CP. Spontaneous BMV MP mutants generated in planta that exhibited CP independent movement were also isolated and analysed. Comparison of the nucleotide differences of the MP genes of BMV-M1, the natural strains and mutants capable of CP-independent movement, together with further mutational analysis of BMV-M1 MP, revealed that single amino acid differences at the C terminus of MP are sufficient to alter the requirement for CP in the movement of BMV-M1. Based on these findings, a possible virus strategy in which a movement mode is selected in plant viruses to optimize viral infectivity in plants is discussed. PMID- 15784915 TI - Cucumber mosaic virus 2a polymerase and 3a movement proteins independently affect both virus movement and the timing of symptom development in zucchini squash. AB - The basis for differences in the timing of systemic symptom elicitation in zucchini squash between a pepper strain of Cucumber mosaic virus (Pf-CMV) and a cucurbit strain (Fny-CMV) was analysed. The difference in timing of appearance of systemic symptoms was shown to map to both RNA 2 and RNA 3 of Pf-CMV, with pseudorecombinant viruses containing either RNA 2 or RNA 3 from Pf-CMV showing an intermediate rate of systemic symptom development compared with those containing both or neither Pf-CMV RNAs. Symptom phenotype was shown to map to two single nucleotide changes, both in codons for Ile at aa 267 and 168 (in Fny-CMV RNAs 2 and 3, respectively) to Thr (in Pf-CMV RNAs 2 and 3). The differential rate of symptom development was shown to be due to differences in the rates of cell-to cell movement in the inoculated cotyledons, as well as differences in the rate of egress of the virus from the inoculated leaves. These data indicate that both the CMV 3a movement protein and the CMV 2a polymerase protein affect the rate of movement of CMV in zucchini squash and that these two proteins function independently of each other in their interactions with the host, facilitating virus movement. PMID- 15784916 TI - Subcellular distribution of mutant movement proteins of Cucumber mosaic virus fused to green fluorescent proteins. AB - The subcellular distribution of the movement proteins (MPs) of nine alanine scanning mutants of Cucumber mosaic virus (CMV), fused to the green fluorescent protein (GFP) and expressed from CMV, was determined by confocal microscopy of infected epidermal cells of Nicotiana tabacum and Nicotiana benthamiana, as well as infected N. benthamiana protoplasts. Only those mutant MPs that were functional for movement in all host species tested localized to plasmodesmata of infected epidermal cells and to tubules extending from the surface of infected protoplasts, as for wild-type CMV 3a MP. Various mutant MPs that were either conditionally functional for movement or dysfunctional for movement did not localize to plasmodesmata and did not form tubules on the surface of infected protoplasts. Rather, they showed distribution to different extents throughout the infected cells, including the cytoplasm, nucleus or the plasma membrane. The CMV 3a MP also did not associate with microtubules. PMID- 15784917 TI - Genotype-level variation in lifetime breeding success, litter size and survival of sheep in scrapie-affected flocks. AB - Five different sheep flocks with natural outbreaks of scrapie were examined to determine associations between individual performance (lifetime breeding success, litter size and survival) and scrapie infection or PrP genotype. Despite different breed composition and forces of infection, consistent patterns were found among the flocks. Regardless of the flock, scrapie-infected sheep produced on average 34 % fewer offspring than non-scrapie-infected sheep. The effect of scrapie on lifetime breeding success appears to be a function of lifespan as opposed to fecundity. Analysis of litter size revealed no overall or genotype differences among the five sheep flocks. Survival, however, depends on the individual's scrapie status (infected or not) and its PrP genotype. Susceptible genotypes appear to perform less well in lifetime breeding success and life expectancy even if they are never affected with clinical scrapie. One possible explanation for these results is the effect of pre-clinical scrapie. Additional evidence supporting this hypothesis is discussed. PMID- 15784918 TI - Clinician-parent communication during informed consent for pediatric leukemia trials. AB - OBJECTIVE: To address the need to describe informed consent in pediatric settings and to identify barriers to parent understanding, this study assessed how aspects of clinician-parent communication during the informed consent conference (ICC) relate to parent understanding of informed consent and parent perception of the impact of the ICC on their anxiety and control. METHODS: Parents of 127 children with newly diagnosed leukemia who were eligible for clinical trials were the participants. The study used comprehensive methods including both observational and self-report assessment methods. RESULTS: Structural equation modeling demonstrated that parent race and socioeconomic status (SES) were powerful predictors of clinician-parent communication, parent anxiety and control as a result of the ICC, and parent understanding. Clinician information giving and partnership building predicted parent participation during the ICC. CONCLUSIONS: These findings may be used to design interventions that increase the effectiveness of the ICC by identifying specific elements of the conference that influence parent affect and understanding. PMID- 15784919 TI - Commentary: SES, ethnicity and goodness-of-fit in clinician-parent communication during pediatric cancer trials. PMID- 15784920 TI - Sociocultural issues in pediatric transplantation: a conceptual model. AB - OBJECTIVE: To demonstrate the value of viewing the pediatric transplant experience through a sociocultural lens and to offer an organized framework for identifying influential sociocultural variables in pediatric transplantation. METHODS: A conceptual model is presented which organizes sociocultural factors that may influence the transplant process. A review of the pediatric and adult transplant literature is conducted. RESULTS: The need for a model addressing sociocultural issues and benefits of using the proposed model is evident. Guided by the proposed model, pediatric psychologists will be prepared to more readily attend to sociocultural influences associated with the transplant experience when conducting research or providing clinical services to patients and families. CONCLUSIONS: Further development and evaluation of the proposed model are necessary to investigate its practical utility and validate the influence of the identified variables on assessment and treatment of pediatric transplant patients as well as patient health outcomes. PMID- 15784921 TI - Very preterm birth is associated with disabilities in multiple developmental domains. AB - OBJECTIVE: Follow-up studies in very preterm children usually present outcome for separate developmental domains. Presence of disabilities in more than one developmental domain will show a more serious outcome picture for extreme preterm infants and may be related to a different degree of perinatal problems. METHODS: At 5.5 years corrected age, outcome in the neurological, motor, cognitive, and behavioral domain was studied in 157 children born < 30 weeks gestation. The children were divided into a normal, a single, or a multiple disability group. Group differences in background, clinical characteristics, and neurodevelopmental outcome at 2 years were evaluated. RESULTS: Thirty-nine percent had a normal developmental outcome, 17% had a single disability, and 44% had multiple disabilities. Multiple disabilities were associated with lower birth weight, BPD, and difficulties according to neurodevelopmental assessments at 2 years. CONCLUSION: Assessments of different developmental domains show that most very preterm children had multiple disabilities. PMID- 15784922 TI - Parent and adolescent versions of the diabetes-specific parental support for adolescents' autonomy scale: development and initial testing. AB - OBJECTIVE: To develop and initially test the psychometric properties of parent and adolescent versions of the Diabetes-Specific Parental Support for Adolescents' Autonomy Scale. METHODS: Data-based scale items were developed, analyzed for content validity, and then piloted with 43 adolescents with type 1 diabetes and their parents. Psychometric properties of the scales were then determined with 100 adolescents with type 1 diabetes and their parents. RESULTS: Content validity indices of .80 or greater were obtained for 26 items. Item analysis in the pilot and large-sample groups resulted in 22 items being deleted. Principal Components Analysis of the remaining four items indicated one factor in both parent and adolescent versions, accounting for 50-62% of variance and with Cronbach alpha coefficients of .67-.80. CONCLUSIONS: This newly developed parsimonious scale, initially tested to be reliable, and valid, will facilitate research on parental support for adolescents' development of diabetes management autonomy. PMID- 15784923 TI - Adjustment to chronic arthritis of childhood: the roles of illness-related stress and attitude toward illness. AB - OBJECTIVE: To examine the relationship of psychosocial stress and attitude toward illness to psychological adjustment among youth with chronic arthritis. METHODS: Seventy-five youths with chronic arthritis aged 8-18 years were administered a semi-structured interview assessing illness-related and nonillness-related stressors in important life domains. Children also completed measures of attitude toward illness, depressive symptoms, and anxiety. Parents completed a measure of child psychosocial adjustment. RESULTS: Higher levels of illness-related and nonillness-related stress were associated with higher levels of anxiety and depressive symptoms and parent-reported adjustment problems, while a more positive attitude toward illness was associated with lower levels of anxiety and depressive symptoms. Attitude toward illness moderated the relationship between stress and depressive symptoms. CONCLUSIONS: Results suggest the importance of assessing life stress and attitude toward illness among youth with arthritis and developing interventions to help children cope with arthritis-related stressors and promote a more positive attitude toward illness. PMID- 15784924 TI - Development of a new fear of hypoglycemia scale: preliminary results. AB - OBJECTIVE: To provide preliminary psychometric data on the Children's Hypoglycemia Index (CHI), a new scale was designed to measure fear of hypoglycemia (FH) in children. METHODS: One hundred and nine children with type 1 diabetes attending a diabetes summer camp completed the CHI, and 57 of these children completed the CHI 2 weeks later. All participants also completed the Children's Hypoglycemia Fear Survey (CHFS), the Revised Children's Manifest Anxiety Scale (RCMAS), and a Hypoglycemia History Form. Parents of the children also completed a Hypoglycemia History Form. RESULTS: The CHI demonstrated good factor structure, internal consistency, test-retest reliability, and criterion related validity with established measures. Frequency of severe hypoglycemic episodes since diabetes diagnosis also predicted scores on the Situation- and General-Fear subscales of the CHI. CONCLUSIONS: The CHI demonstrated reliability and validity as a measurement of children's FH. The use of this measure offers important clinical and research implications. PMID- 15784926 TI - DO questions evidence for including tobacco dependence curricula. PMID- 15784925 TI - Brief report: a qualitative study of gender preferences and motivational factors for physical activity in underserved adolescents. AB - OBJECTIVE: This study used focus groups to identify preferences for physical activity (PA) and motivational themes for increasing PA in underserved adolescents. METHODS: Fifty-one adolescents, on the free lunch program (25 males and 26 females; ages 10-13 years; 72% African-American, 19% white, and 9% other minorities) participated in focus groups (6-10 children per group; same gender). The data were transcribed and coded into themes. A content analysis was performed using QSR NVivo. RESULTS: Boys rated their favorite activities as playing basketball, football, soccer, and baseball. Girls rated their favorite activities as playing basketball, swimming, and roller-skating. For motivational themes, boys and girls most frequently stated that they would participate in PA if it was a fun, provided a health benefit, and if there were a variety of choices offered. Being involved with friends was also identified as important to both boys and girls. CONCLUSIONS: The findings from this qualitative study suggest that including components of choice and a variety of physical activities may be important elements for intervention programs in underserved adolescents. PMID- 15784927 TI - Chairman of COPT concludes debate on "hardship exception". PMID- 15784928 TI - Intramuscular ketorolac versus osteopathic manipulative treatment in the management of acute neck pain in the emergency department: a randomized clinical trial. AB - Ketorolac tromethamine injected intramuscularly (IM) has been shown to be an effective analgesic in treating patients with acute musculoskeletal pain in the emergency department (ED). The authors compare the efficacy of a single dose of IM ketorolac to osteopathic manipulative treatment (OMT) as delivered in the ED for the management of acute neck pain. A randomized clinical trial was conducted in three EDs. A convenience sample of 58 patients with acute neck pain of less than three weeks' duration were enrolled. Subjective measures of pain intensity on an 11-point numerical rating scale were gathered from patients immediately before treatment and one hour afterward. Subjects received either OMT or 30 mg, IM ketorolac. Subjects' perceived pain relief was also recorded at one hour after treatment on a subjective 5-point pain relief scale. Twenty-nine patients received IM ketorolac, and 29 patients received OMT. Although both groups showed a significant reduction in pain intensity, 1.7+/-1.6 (P <.001 [95% CI, 1.1-2.3]) and 2.8+/-1.7 (P <.001 [95% CI, 2.1-3.4]), respectively, patients receiving OMT reported a significantly greater decrease in pain intensity (P=.02 [95% CI, 0.2 1.9]). When comparing pain relief at one hour posttreatment, there was no significant difference between the OMT and ketorolac study groups (P=.10). The authors found that, at one hour posttreatment, OMT is as efficacious as IM ketorolac in providing pain relief and significantly better in reducing pain intensity. The authors conclude that OMT is a reasonable alternative to parenteral nonsteroidal anti-inflammatory medication for patients with acute neck pain in the ED setting. PMID- 15784929 TI - Evaluating the patient with peripheral nervous system complaints. AB - Patients commonly seek care from their family physicians for symptoms that are suggestive of peripheral nervous system (PNS) dysfunction. At some point in active practice, virtually all family physicians will be required to conduct at least an initial evaluation of a patient with PNS. The authors outline and describe common themes found in the symptoms and diagnoses of PNS dysfunctions. These themes may be useful to physicians when performing initial evaluations of patients with PNS dysfunctions. The authors also discuss diagnostic methods and effective therapeutic interventions for this population. PMID- 15784930 TI - Cellular telephones and electronic communication patterns among families in metropolitan New York. AB - OBJECTIVE: The authors gathered survey data regarding the use of cellular telephones among families who reside in the New York metropolitan area and whose children receive medical care in a hospital-affiliated general pediatrics clinic. METHODS: Two investigators distributed a 34-question, self-administered survey in pencil-and-paper format to pediatric patients and the adults who accompanied them in the waiting room of a hospital-affiliated, outpatient pediatric practice in Mineola, New York. RESULTS: Completed surveys were obtained from 35 families. Twenty-eight (80%) of the 35 families completing surveys reported that at least one family member owned and used a cellular telephone. Of these 28 families, 9 (32%) reported that at least one household member aged to 18 years owned and used a cellular telephone. The mean age at which children obtained their first cellular telephone was 15.3 years. The primary reason these respondents cited for providing children with cellular telephones was safety (75%). CONCLUSION: The authors conclude that cellular telephone use is widespread among families in the New York metropolitan area. Although the primary reason families provide for obtaining cellular telephones is safety, convenience to household members and peer pressure also play roles. PMID- 15784934 TI - The importance of being statistical. PMID- 15784933 TI - Quantitative analysis of severe acute respiratory syndrome (SARS)-associated coronavirus-infected cells using proteomic approaches: implications for cellular responses to virus infection. AB - We present the first proteomic analysis on the cellular response to severe acute respiratory syndrome-associated coronavirus (SARS-CoV) infection. The differential proteomes of Vero E6 cells with and without infection of the SARS CoV were resolved and quantitated with two-dimensional differential gel electrophoresis followed by ESI-MS/MS identification. Moreover isotope-coded affinity tag technology coupled with two-dimensional LC-MS/MS were also applied to the differential proteins of infected cells. By combining these two complementary strategies, 355 unique proteins were identified and quantitated with 186 of them differentially expressed (at least 1.5-fold quantitative alteration) between infected and uninfected Vero E6 cells. The implication for cellular responses to virus infection was analyzed in depth according to the proteomic results. Thus, the present work provides large scale protein-related information to investigate the mechanism of SARS-CoV infection and pathogenesis. PMID- 15784935 TI - Publishing without ethical approval. PMID- 15784936 TI - Mandibular incisal edge demineralization and caries associated with Twin Block appliance design. AB - Demineralization and caries are well documented and unwanted complications of orthodontic therapy. These are most commonly reported in relation to fixed appliance therapy. Five cases are presented of patients undergoing Twin Block appliance therapy with the appliances incorporating lower labial segment capping, illustrating a pattern of demineralization and caries of the incisal tips of the mandibular labial segment, influenced by the incisal capping and the frequent intake of carbonated soft drinks. PMID- 15784937 TI - Canine transposition following trauma and loss of a central incisor: treatment options. AB - The aim of this article is to make clinicians aware of the possibility of canine transposition following loss of the upper central incisor. Using two case reports it considers the opportunities for rehabilitation of the occlusion, and discusses surgical, orthodontic and restorative considerations. PMID- 15784938 TI - Combined cephalometric and stent planning for palatal implants. AB - Several aspects of palatal implant usage are technique sensitive. In particular, problems during the insertion stage may compromise implant osseo-integration, or its subsequent ease of handling and effectiveness. This paper describes a systematic approach to combined cephalometric and model planning, and subsequent stent fabrication for Orthosystem palatal implants. The authors recommend this protocol in order to optimize three-dimensional control of implant positioning, and to both simplify and standardize the insertion stage. PMID- 15784939 TI - A randomized controlled trial comparing the quadhelix and the expansion arch for the correction of crossbite. PMID- 15784940 TI - A 100th anniversary: Sandstedt's experiments on tissue changes during tooth movement. AB - In 2004, it is 100 years since the experiments by Carl Sandstedt on tissue changes during orthodontic tooth movement were published in an international journal, shortly after his early death. PMID- 15784941 TI - A randomized controlled trial comparing the quadhelix and the expansion arch for the correction of crossbite. AB - OBJECTIVE: To compare the use of the quadhelix and the expansion arch for the correction of crossbite. DESIGN: A prospective randomized clinical trial supported by preliminary laboratory measurements. The null hypothesis was that there was no difference in the clinical effectiveness of the two expansion devices in terms of crossbite correction. SETTING: Queen's Hospital, Burton on Trent and The University of Birmingham, School of Dentistry. PARTICIPANTS: The first 60 patients on the orthodontic waiting list at Queen's Hospital who required expansion of the maxillary arch as part of the treatment plan were allocated to be treated with either a quadhelix or an expansion arch by random allocation. Twenty-eight and 27 members of each respective group completed the study. MATERIALS: Commercial quadhelix arches (3M Unitek) and custom-made expansion arches METHODS: The force produced by the type of expansion arches used in the study was measured in the laboratory to be 1.8 N at 10 mm of expansion. Quadhelix arches of sizes 2 and 3 were found to produce equivalent forces at 5 and 7 mm of expansion respectively. Either expansion device was fitted to the 60 participants according to random allocation and expanded by the standard amount. Intermolar and intercanine expansion was measured after 4, 8 and 12 weeks. Patient opinion was assessed by using a questionnaire. RESULTS: The quadhelix and the expansion arch were equally effective in producing expansion (p>0.05). After 12 weeks, the two types of archwire had produced mean intermolar expansions of 4.54 and 5.09 mm and intercanine expansions of 1.41 and 2.12 mm, respectively. Both types of arch were reported as uncomfortable by a majority of patients, the quadhelix affected mainly the tongue and the expansion arch the cheeks. The appearance of the quadhelix was disliked by 25% of participants, while 70% disliked the expansion arch. CONCLUSIONS: The null hypothesis was confirmed. However, the expansion arch had several advantages that made it a cheap alternative to the quadhelix for crossbite expansion, because it can be made and fitted at the chairside. PMID- 15784943 TI - How to avoid common errors in clinical photography. AB - This paper demonstrates some of the errors commonly seen in both conventional and digital photography when used for clinical purposes, and details how some of these mistakes may be avoided. PMID- 15784942 TI - Chlorhexidine-modified glass ionomer for band cementation? An in vitro study. AB - OBJECTIVE: To compare the mean retentive strength, predominant site of band failure, amount of cement remaining on the tooth at deband and survival time of orthodontic micro-etched bands cemented with chlorhexidine-modified (CHXGIC) or conventional glass ionomer cement (GIC). DESIGN: In vitro study. SETTING: Dental Materials Laboratory. MATERIALS AND METHODS: One-hundred-and-twenty intact, caries-free third molars were collected from patients attending for third molar surgery. These were stored for 3 months in distilled water and decontaminated in 0.5% chloramine. To assess retentive strength, 80 teeth were randomly selected and 40 were banded with each cement. Testing was undertaken using a Nene M3000 testing machine at a cross-head speed of 1 mm/min. Following debanding, the predominant site of failure was recorded as cement-enamel or cement-band interface. The amount of cement remaining on the tooth surface following deband was assessed and coded. Survival time for another 40 banded specimens, 20 cemented with each cement, was assessed following application of mechanical stress in a ball mill. MAIN OUTCOME MEASURES: Retentive strength, predominant site of failure, amount of cement remaining on the tooth surface, survival time. RESULTS: Mean retentive strength for bands cemented with CHXGIC (0.32 MPa, SD 0.09) or GIC (0.28 MPa, SD 0.07) did not differ significantly (p=0.05). All bands failed at the enamel-cement interface. There was no significant difference in the amount of cement remaining on the tooth surface after deband for each cement type (p=0.23). The mean survival time of bands cemented with CHXGIC or GIC was 7.0 and 6.4 hours, respectively (p=0.23). CONCLUSIONS: There was no significant difference in mean retentive strength, amount of cement remaining on the tooth after deband or mean survival time of bands cemented with CHXGIC or GIC. Bands cemented with either cement failed predominantly at the enamel-cement interface. The results suggest that CHXGIC may have comparable clinical performance to GIC for band cementation. PMID- 15784945 TI - Current products and practices: personal digital assistants in orthodontics. AB - Personal Digital Assistants (PDAs) have certain advantages over conventional diaries and are thought of as the 'electronic filofax'. They can be backed up, used for personal and professional organization, integrated with mobile phones for email and Internet access, image storage, logbook functions and audit. They are limited when used for desktop functions. This article attempts to highlight the current advantages and disadvantages of PDAs for use in orthodontics, and provides some guidance to those who are considering the purchase of such a device. PMID- 15784947 TI - Effects of adaptation to altered display gain on the control of single aimed movements. AB - A display gain setting defines the mapping of a movement to the real-time visual display. In two experiments we investigated how the acquired adaptation to low and high display gain affected motor control of single aimed stylus movements. Experiments differed with respect to how gain was varied. In Experiment 1, gain was realized by manipulating the surface displacements with a constant display. The results show an expected linear decrease of movement time and spatial accuracy, which is in sharp contrast with the often-reported U-shaped relation. Experiment 2 was run to study the influence of visual feedback in low- and high gain conditions. The manipulation of gain was realized by display variation with unaltered surface displacements. The linear increase of movement time and feedback processing with gain and the unaltered spatial accuracy across conditions, suggested that participants actively adapted to the displayed visual information in altered gain conditions. PMID- 15784948 TI - Finger-tapping ability in male and female pianists and nonmusician controls. AB - Using fast tapping tasks with each of the four fingers (single-finger tapping) and with two of the fingers used alternately (double-finger tapping), the ability to make rapid tapping movement by the individual fingers was compared between expert pianists and nonmusician controls in both genders. Maximal pinch and grasp forces were also measured to assess strength of individual fingers and whole hand, respectively. Movement of the ring and little fingers was slower than that of the index and middle fingers in both the pianists and controls. The slowness of the ring and little fingers was, however, much less evident in the pianists than the controls in both tapping tasks. The pianists also had smaller intertap interval variability for the index and middle fingers. No pianist-control difference was found for the pinch and grasp forces. Piano training, therefore, effectively changed the ability to move individual fingers rapidly, but not their flexor strength. No gender difference was found in any of the tapping tasks though males had greater strength. Gender thus does not appear to be a factor differentiating the ability to move individual fingers rapidly. PMID- 15784949 TI - Differential transfer processes in incremental visuomotor adaptation. AB - Visuomotor adaptive processes were examined by testing transfer of adaptation between similar conditions. Participants made manual aiming movements with a joystick to hit targets on a computer screen, with real-time feedback display of their movement. They adapted to three different rotations of the display in a sequential fashion, with a return to baseline display conditions between rotations. Adaptation was better when participants had prior adaptive experiences. When performance was assessed using direction error (calculated at the time of peak velocity) and initial endpoint error (error before any overt corrective actions), transfer was greater when the final rotation reflected an addition of previously experienced rotations (adaptation order 30 degrees rotation, 15 degrees, 45 degrees) than when it was a subtraction of previously experienced conditions (adaptation order 45 degrees rotation, 15 degrees, 30 degrees). Transfer was equal regardless of adaptation order when performance was assessed with final endpoint error (error following any discrete, corrective actions). These results imply the existence of multiple independent processes in visuomotor adaptation. PMID- 15784950 TI - Muscle synergies at the elbow in static and oscillating isometric torque tasks with dual degrees of freedom. AB - This study's aim was to identify the effect of oscillation of torques in isometric tasks under identical mechanical conditions on the muscle synergies used. It was hypothesized that bi-functional muscles would play a lesser role in torque oscillation, because they would also generate an undesired oscillation. Thus, changes in muscle synergies were expected as a consequence of oscillation in torque generation. The effect of the trajectory of torque generation was investigated in dual-degrees-of-freedom submaximal isometric oscillation torque tasks at the elbow. The torques were flexion-extension and supination-pronation. Oscillation torques were compared with static torque generations at four torque positions during oscillation. Muscle activity was determined with surface electromyography. Compared with the static torque tasks, the oscillation tasks showed an overall increased muscle activity. The oscillation tasks, however, showed similar activity patterns and muscle synergies compared to the static composite tasks. It was found that the motor system is well able to control different orthogonal combinations of slow torque oscillations and constant torques by employing a single oscillating muscle synergy. PMID- 15784951 TI - The effect of workspace on the use of motor abundance. AB - The uncontrolled manifold approach was used to examine the effect of workspace location on the use of motor abundance to control the hand's path during reaching. Participants pointed to targets located in the contralateral and ipsilateral workspaces at two different distances. When reaching to all parts of the workspace, the component of joint configuration variance consistent with an identical hand path across trials was significantly higher than the component of joint configuration variance leading to a variable hand path. The relative magnitude of this difference was affected primarily by target orientation and minimally by target distance. The control of hand-path direction when reaching ipsilaterally was associated with more selective use of motor abundance compared to reaching contralaterally. The control of hand-path extent was not affected by target orientation. Biomechanical factors are discussed as possible reasons that lead to the observed selective workspace effects. PMID- 15784952 TI - Energy-minimization bias: compensating for intrinsic influence of energy minimization mechanisms. AB - Anecdotal and scientific evidence suggest humans tend to undershoot targets in rapid movements. We investigated whether this undershoot bias derives from energy minimization mechanisms. Participants performed 200 trials of two tasks: (1) a simple slider push to a target, and (2) a modified version of (1), designed so overshooting was less energy consuming than undershooting. Results support that the undershoot bias found in (1), as well as the overshoot bias found in (2), results from an energy minimization mechanism. Energy minimization might be inherent to biological systems. Movement biases were un desirable for maximal performance. Nonetheless, participants presented biases despite financial incentives to perform maximally. Participants did, however, appear sensitive to systematic errors produced by the attraction to less energy costly responses. We suggest that the motor system is constrained such that maximal performance trades off with energetic optimality although humans are able to learn and compensate for the energy minimization biases. PMID- 15784953 TI - Selenium level in benign and cancerous prostate. AB - The dietary microelement selenium (Se) has been proposed as a potential chemopreventive agent for prostate cancer. This element is present in various amounts in all tissues. Little information is available on Se level in patients with prostate gland disorders. The levels of Se in prostatic gland of patients with prostate cancer, benign prostate hyperplasia, and healthy controls were examined. The Se level for benign prostate hyperplasia (156 +/- 30.6 ng/g) was the same as in the control group (157 +/- 26.0 ng/g), but in the gland of prostate cancer patients (182 +/- 34.1 ng/g wet weight), the Se level was significantly (p < 0.01) higher than in both healthy controls and benign prostate hyperplasia. Thus, the Se level in human healthy controls is lower than in kidney and liver but higher compared with other tissues. PMID- 15784954 TI - Beneficial effects of selenium on some enzymes of diabetic rat heart. AB - It is known that selenium compounds can restore some metabolic parameters in experimental diabetes. However, as there are no clear data about their effects on the altered antioxidant defense system of the diabetic heart, we aimed to investigate whether these beneficial effects extend to the alterations of some enzyme activities, which play important roles in antioxidant defense system. Diabetes was induced by streptozotocin (50 mg/kg body weight) and rats were then treated with sodium selenite (5 micromol/kg/d) for 4 wk. Sodium selenite treatment of the diabetic rats significantly restored the altered activities of glutathione-S-transferase, glucose-6-phosphate dehydrogenase, and 6 phosphogluconate dehydrogenase, which are involved in the glutathione metabolism of the heart, but slightly but significantly decreased the high blood glucose level. In summary, the present study suggests that the beneficial effects of sodium selenite treatment appears to be the result of the restoration altered activities of the antioxidant enzymes in diabetic heart tissue. PMID- 15784955 TI - Increases of calcium and magnesium and decrease of iron in human posterior longitudinal ligaments of the cervical spine with aging. AB - To elucidate compositional changes of ligaments with aging, the authors investigated age-related changes of elements in the posterior longitudinal ligaments (PLLs) by inductively coupled plasma-atomic emission spectrometry. After the ordinary dissection, PLLs were resected from the subjects ranging in age from 65 to 95 yr. The PLLs of the cervical spine were resected between the fourth and fifth cervical vertebrae, the PLLs of the thoracic spine between the fifth and seventh thoracic vertebrae, and the PLLs of the lumbar spine between the second and third lumbar vertebrae. Calcium and magnesium increased progressively with aging in the PLLs of the cervical spine, but they did not increase with aging in the PLLs of the thoracic and lumbar spine. In contrast, iron decreased gradually with aging in the PLLs of the cervical spine. Regarding the relationships among elements, significant correlations were found among the contents of calcium, phosphorus, magnesium, and sodium in the PLLs of the cervical spine. PMID- 15784956 TI - Copper increases the damage to DNA and proteins caused by reactive oxygen species. AB - Copper [Cu(II)] is an ubiquitous transition and trace element in living organisms. It increases reactive oxygen species (ROS) and free-radical generation that might damage biomolecules like DNA, proteins, and lipids. Furthermore, ability of Cu(II) greatly increases in the presence of oxidants. ROS, like hydroxyl (.OH) and superoxide (.O(2)) radicals, alter both the structure of the DNA double helix and the nitrogen bases, resulting in mutations like the AT-->GC and GC-->AT transitions. Proteins, on the other hand, suffer irreversible oxidations and loss in their biological role. Thus, the aim of this investigation is to characterize, in vitro, the structural effects caused by ROS and Cu(II) on bacteriophage lambda DNA or proteins using either hydrogen peroxide (H(2)O(2)) or ascorbic acid with or without Cu(II). Exposure of DNA to ROS-generating mixtures results in electrophoretic (DNA breaks), spectrophotometric (band broadening, hypochromic, hyperchromic, and bathochromic effects), and calorimetric (denaturation temperature [T(d)], denaturation enthalpy [DeltaH], and heat capacity [C(p)] values) changes. As for proteins, ROS increased their thermal stability. However, the extent of the observed changes in DNA and proteins were distinct, depending on the efficiency of the systems assayed to generate ROS. The resulting effects were most evident when Cu(II) was present. In summary, these results show that the ROS, .O2 and .OH radicals, generated by the Cu(II) systems assayed deeply altered the chemical structure of both DNA and proteins. The physiological relevance of these structural effects should be further investigated. PMID- 15784957 TI - Effects of a combination of niacin and chromium(III)-chloride on the skin and lungs of hyperlipemic rats. AB - The aim of this study was to investigate the biochemical effects of niacin and chromium(III)-chloride on serum lipid peroxidation, uric and sialic acids, and the extent of lipid peroxidation and glutathione levels in skin and lung tissues of hyperlipidemic rats. In this study, female Swiss albino rats, 12 mo old, were used. They were randomly divided into four groups. Group I animals were fed with a standard pellet diet and water ad libitium. Group II rats were fed with a standard pellet diet and were treated with a dose of 250 microg/kg body weight CrCI(3).6H(2)O and 100 mg/kg body weight niacin, for 45 d, by the gavage technique. Group III rats were fed a lipogenic diet in which 2% cholesterol, 0.5% cholic acid, and 20% sunflower oil were added to the pellet chow. In addition, the animals in this group drank water containing 3% ethanol. This regime was maintained for 60 d. The rats in group IV were maintained in the same food and drink regime as the animals in group III. After 2 wk, the animals showed symptoms of hyperlipemia and they were treated with 250 microg/kg body weight CrCI(3).6H(2)O and 100 mg/kg body weight niacin, by gavage, for 45 d. On d 60, the blood and the skin and lungs samples were taken from animals. In the hyperlipemic groups, a reduction of the lung glutathione level and an increase in serum, lung, and skin lipid peroxidation levels and in serum sialic and uric acid were observed. In rats treated with a combination of niacin and Cr(III), the skin and serum lipid peroxidation and the sialic and uric acid levels decreased while showing an increase of lung glutathione activity. These results suggest that niacin and Cr(III), when administered in combination, have a protective effect against skin and lung tissues damage as a result of hyperlipidemia. PMID- 15784958 TI - Determination of mercury in hair by square-wave anodic stripping voltammetry at a rotating gold disk electrode after microwave digestion. AB - A simple and reliable method for the determination of mercury in hair on a rotating gold disk electrode using subtractive anodic stripping voltammetry without removal of oxygen is reported. Voltammetric and microwave parameters were optimized to obtain the best analytical results. Parameters such as supporting electrolyte concentration, influence of chloride in the Hg peak, deposition potential, scan rate, accumulation time, rotation rate, square-wave amplitude, and electrode conditioning were studied. Pressurized microwave-assisted digestion of hair, suitable for the accurate voltammetric determination of Hg, was evaluated using six acid mixtures and several time-power programs. Under the optimized conditions, no interference by copper, cadmium, lead, nickel, manganese, iron, or zinc was found at concentrations corresponding to their occurrence in normal hair. A calibration plot between 6,67 and 46,69 microg/L was linear, with r(2) better than 0.999. The detection limit for a deposition time of 60 s at 254 g was calculated as 1.92 nM (3omega). Validation of the method was demonstrated with the use of a certified reference sample of hair. Eight real samples of hair (four unexposed children and four exposed persons) were also analyzed. PMID- 15784959 TI - Characterization of hypoglycemiant plants by total reflection X-ray fluorescence spectrometry. AB - In this work, synchrotron radiation total reflection X-ray fluorescence spectrometry (SRTXRF) was used to determine trace elements in eight hypoglycemiant plants (Trigonella foenum graecum, Panax ginseng, Pfaffia paniculata, Myrcia speciosa, Zea mays, Harpagophytum procumbens, Syzygium jambolona, and Bauhinia forficate). The elements P, K, Ca, Ti, Mn, Fe, Cu, Zn, Rb, and Sr were detected in all medicinal plants investigated, whereas Si, S, Sc, V, Cr, Co, Ni, Se, Nb, Mo, Sn, Sb, Ba, Hg, and Pb were detected only in some of the samples. The concentration of elements in hypoglycemiant plants varied from 0.15 microg/g of Co to 3.0 x 10(4) microg/g of K and the mean of experimental limit of detection for these elements were 0.14 and 3.6 microg/g, respectively. PMID- 15784960 TI - Genetic association of BACE1 gene polymorphism C786G with late-onset Alzheimer's disease in Chinese. AB - Beta-amyloid (Abeta) peptides are derived from the endoproteolytic processing of amyloid precursor protein (APP) and play a key role in the pathogenesis of Alzheimer's disease (AD). Beta-site APP-cleaving enzyme 1 ([BACE1] also known as beta-secretase) is responsible for cleaving APP to generate neurotoxic Abeta peptides in patients with AD. The BACE1 gene is located on chromosome 11q23.3, near the recently identified region with increased lod scores for AD. The biological functional and genetic association studies indicated that the BACE1 gene might be a genetic risk factor for late-onset Alzheimer's disease (LOAD). To investigate an association between the BACE1 C786G polymorphism and sporadic LOAD in Chinese, we examined 105 LOAD patients and 130 healthy controls. Our results showed higher frequency of the 786G-allele in LOAD patients (38.6%) than that in controls (28.5%), and a statistical significance was observed for an association of the G-allele with LOAD (odds ratio [OR] = 1.58, 95% confidence interval [CI] 1.07-2.23, p = 0.02). We also found a synergetic interaction between the G-allele and apolipoprotein E allele 4 (APOE e4) status on the risk of LOAD (OR = 1.91, 95% CI 1.23-2.95, p = 0.003). These results suggest that BACE1 gene polymorphism C786G might act as an APOE epsilon4 allele-dependent risk factor for developing LOAD in Chinese. PMID- 15784961 TI - Roles of Ras-Erk in apoptosis of PC12 cells induced by trophic factor withdrawal or oxidative stress. AB - To understand the role of Ras-MAPK (mitogen-activated protein kinase) in trophic factor withdrawal- and oxidative stress-induced apoptotic cell death processes, undifferentiated rat pheochromocytoma PC12 cells and a PC12 variant cell line stably expressing the Ras dominant-negative mutant (M-M17-26) were subjected to serum withdrawal in the absence or presence of H2O2 treatment. The extent of cell death was analyzed by lactate dehydrogenase release, internucleosomal DNA fragmentation, and caspase-3 assays. Both serum withdrawal and H2O2 treatment induced apoptotic cell death in PC12 cells, and the extent of cell death was greatly enhanced in M-M17-26 cells. DNA fragmentation induced by serum withdrawal or H2O2 treatment was blocked completely by a general caspase inhibitor, Z-VAD FMK. A selective MAPK kinase inhibitor, U0126, blocked the H2O2-induced phosphorylation of Erk1/2 (extracellular signal-regulated kinase) in PC12 cells and increased the levels of active caspase-3 in M-M17-26 under serum withdrawal or H2O2 treatment. In addition, the short-term H2O2 treatment (5-30 min) was sufficient to cause DNA fragmentation in M-M17-26 cells even though H2O2 was removed and cells were incubated in regular growth medium with complete serum for 24 h. However, similar, short-term H2O2 treatment of PC12 cells did not induce DNA fragmentation 24 h later. These results suggest that the Ras-Erk pathway is critical in mediating protection against apoptotic cell death induced by either trophic factor withdrawal or increased oxidative stress. PMID- 15784962 TI - Characterization of the signaling pathway downstream p75 neurotrophin receptor involved in beta-amyloid peptide-dependent cell death. AB - The accumulation of beta-amyloid (Abeta) peptide is a key pathogenic event in Alzheimer's disease. Previous studies have shown that Abeta peptide can damage neurons by activating the p75 neurotrophin receptor (p75NTR). However, the signaling pathway leading to neuronal cell death is not completely understood. By using a neuroblastoma cell line devoid of neurotrophin receptors and engineered to express either a full-length or a death domain (DD)-truncated form of p75NTR, we demonstrated that Abeta peptide activates the mitogen-activated protein kinases (MAPKs) p38 and c-Jun N-terminal kinase (JNK). We also found that Abeta peptide induces the translocation of nuclear factor-kappaB (NF-kappaB). These events depend on the DD of p75NTR. Beta-amyloid (Abeta) peptide was found not to be toxic when the above interactors were inhibited, indicating that they are required for Abeta-induced neuronal cell death. p75 neurotrophin receptor (p75NTR)-expressing cells became resistant to Abeta toxicity when transfected with dominant-negative mutants of MAPK kinases 3, 4, or 6 (MKK3, MKK4, or MKK6), the inhibitor of kappaBalpha, or when treated with chemical inhibitors of p38 and JNK. Furthermore, p75NTR-expressing cells became resistant to Abeta peptide upon transfection with a dominant-negative mutant of p53. These results were obtained in the presence of normal p38 and JNK activation, indicating that p53 acts downstream of p38 and JNK. Finally, we demonstrated that NF-kappaB activation is dependent on p38 and JNK activation. Therefore, our data suggest a signaling pathway in which Abeta peptide binds to p75NTR and activates p38 and JNK in a DD dependent manner, followed by NF-kappaB translocation and p53 activation. PMID- 15784963 TI - Peripheral sensory neurons survive in the absence of alpha- and gamma-synucleins. AB - Physiological functions of alpha-synuclein, a protein implicated in certain types of neurodegeneration, and two other members of the same family, beta-synuclein and gamma-synuclein, are not clearly understood. It has been suggested that synucleins are involved in intracellular processes associated with survival of neurons and their response to stress, and that changes of synuclein ratio might have deteriorating effects on neurons. In wild-type mice, sensory neurons of the peripheral nervous system express alpha-synuclein and notably high levels of gamma-synuclein, but targeted inactivation of either of these genes has no effect on these neurons. Here we produced double, alpha-synuclein/gamma-synuclein null mutant mice, which develop normally, are fertile, and show no obvious signs of pathology in adulthood. Survival of alpha/gamma-synuclein-deficient peripheral sensory neurons in vivo and in primary tissue culture is indistinguishable from survival of wild-type neurons. The absence of two synucleins does not lead to expression in sensory neurons of the third member of the family, beta-synuclein. Therefore, our results demonstrate that neurons with normally high levels of synuclein(s) can develop and survive normally in the absence of any of these proteins. This suggests that other intraneuronal mechanisms and pathways effectively compensate the loss of synuclein function in null mutant animals. PMID- 15784964 TI - Humanin attenuates apoptosis induced by DRPLA proteins with expanded polyglutamine stretches. AB - Dentatorubral-pallidoluysian atrophy (DRPLA) is an autosomal-dominant neurodegenerative disorder caused by expansion of CAG repeats in the DRPLA gene, which codes for a polyglutamine (polyQ) stretch. The expanded polyQs are known to form intracellular aggregates and to confer neurotoxic activity. Recent studies have indicated that activation of apoptosis signal-regulating kinase 1 (ASK1) is involved in polyQ-induced apoptosis. Humanin (HN) is an endogenous peptide that inhibits neuronal cell death caused by mutant Alzheimer's disease genes, and this neuroprotective factor has recently been reported to suppress apoptosis by inhibiting activation of ASK1. To test the anti-ASK1 effect of HN on polyQ neurotoxicity, we constructed neuronal PC12 cells expressing expanded polyQs under the control of the Tet-Off system. Using this cell line, we showed that HN suppresses apoptotic cell death induced by expanded polyQs. However, the suppression was incomplete, suggesting that polyQs also stimulate other pathogenic cascades unrelated to ASK1. We further showed that HN suppresses polyQ aggregate formation. This result implied the possibility that aggregation is also related to the polyQ-mediated cascade involving ASK1 activation. Although the details remain uncertain, our results suggest that ASK1 is potentially involved in pathogenesis of DRPLA and that HN might contribute partially to the suppression of neurodegeneration in polyQ diseases. PMID- 15784965 TI - Effects of acute amphetamine administration on AMPA-mediated synaptic activity and expression of AMPA receptor subunit 2 of brain neurons. AB - We investigated the role of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor (AMPAR) in mechanisms underlying the action of amphetamine (Amph) on brain neurons, for AMPAR has been proposed to participate in psychotic and neurodegenerative disorders. In the cultured rat brain cortical neurons pretreated with 1 microM Amph for 1 h, the accumulation of 45Ca2+ driven by 10 min incubation with 100 microM AMPA was reduced by about 36%. This Amph-induced decrease seems to involve L-type voltage-gated Ca2+ channels, because the AMPA induced 45Ca2+ uptake was blocked by 70% and 80%, respectively, for untreated and Amph-treated neurons in the presence of nifedipine (1 microM), an antagonist to L type calcium channels. Whole-cell, patch-clamp recording revealed that AMPA elicited current amplitude became 26% lower than the control in Amph-treated cultured neurons. Moreover, Amph treatment down-regulated the level of flip-form glutamate receptor 2 (GluR2) mRNA by 27% in cultured neurons but did not change the expression of GluR2 proteins and flop-form mRNA, as detected by quantitative immunocytochemistry and in situ hybridization. In contrast, in postnatal day 4 rats at 1 h after receiving one intraperitoneal injection of 5 mg/kg of Amph, levels of flip GluR2 mRNA were up-regulated by 13% and 18% in neurons of motor cortex layer 5 and pyramidal neurons of hippocampal CA3, respectively. The data suggest that acute action of Amph on brain neurons is possibly associated with decreased AMPA-mediated Ca2+ influx and current amplitude, as well as modified expression of the GluR2 mRNA. PMID- 15784966 TI - Recombinant erythropoietin down-regulates IL-6 and CXCR4 genes in TNF-alpha treated primary cultures of human microvascular endothelial cells: implications for multiple sclerosis. AB - In multiple sclerosis (MS), disruption of the blood-brain barrier might lead to new gadolinium-enhanced lesion formation in the brain and cause acute relapses. Current therapeutic options for acute relapses in MS are limited. The effect of recombinant erythropoietin (rEPO) on cytokine gene expression in TNF-alpha treated human brain microvascular endothelial cells was studied. The cells were controls (untreated), exposed for either 6 or 24 h to TNF-alpha or TNF alpha/rEPO. Of the 96 genes studied, interleukin-6 (IL-6), IL-1beta, CXCR4, and IL-1alpha genes were down-regulated when treated with TNF-alpha/rEPO for 6 h as compared with TNF-alpha alone. At 24 h, IL-6 and CXCR4 gene expression was 4.24 and 2.98, respectively. Quantitative RT-PCR analysis showed down-regulation by 3.86 and 1.9 for IL-6 and CXCR4 genes, respectively. Our findings suggest that further studies are warranted to evaluate the use of EPO in minimizing acute relapses in MS. PMID- 15784967 TI - Trafficking of adenosine A2A and dopamine D2 receptors. AB - An interaction between adenosine A2A and dopamine D2 receptors has been demonstrated previously. It is generally found that agonist treatment internalizes receptors, including A2A and D2, whereas less is known of the long term effects involved in the agonist-mediated trafficking of A2A and D2 receptors. Furthermore, the possible influence of the antagonists on receptor trafficking is still undefined. The present studies focus on the long-term effects of A2A and D2 agonist and D2 antagonist treatments on both A2A and D2 receptor trafficking studied at three different time intervals--3, 15, and 24 h. In addition, with the fluorescence resonance energy transfer technique, formation of heteromeric A2A and D2 receptor complexes was shown in the cotransfected CHO cell line. Confocal microscopy analysis showed that a 3-h treatment with the D2 agonist induced coaggregation of A2A/D2 receptors. These A2A/D2 receptor coaggregates internalized after 15 h with a recruitment of the receptors back to the cell membrane after 24 h. In contrast to the effects of the agonist treatment, a 3-h treatment with the D2-like antagonist raclopride increased both A2A and D2 immunoreactivity, indicating that the D2 antagonist stabilizes the D2 receptor and thereby reduces the internalization of both of the A2A and D2 receptors. Taken together, an activation of either A2A and D2 receptor or blockade of D2 receptors will cause long-lasting changes in A2A and D2 receptor trafficking. PMID- 15784968 TI - AdpA, a central transcriptional regulator in the A-factor regulatory cascade that leads to morphological development and secondary metabolism in Streptomyces griseus. AB - A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) is a microbial hormone that triggers aerial mycelium formation and secondary metabolism in Streptomyces griseus. A-factor produced in a growth-dependent manner switches on the transcription of adpA encoding a transcriptional activator by binding to the A-factor receptor protein (ArpA), which has bound the adpA promoter, and dissociating the DNA-bound ArpA from the DNA. AdpA then activates a number of genes with various functions required for morphological development and secondary metabolism, forming an AdpA regulon. AdpA, which contains a ThiJ/PfpI/DJ-1-like dimerization domain at its N-terminal portion and an AraC/XylS-type DNA-binding domain at its C-terminal portion, is a representative of a large subfamily of the AraC/XylS family. AdpA binds various positions with respect to the transcriptional start points of the target genes and recruits RNA polymerase to the specific promoter region, and facilitates the isomerization of the RNA polymerase-DNA complex into an open complex competent for transcriptional initiation. The AdpA-binding consensus sequence is 5'-TGGCSNGWWY-3' (S: G or C; W: A or T; Y: T or C; N: any nucleotide). The DNA-binding specificity of AdpA in conjunction with that of other AraC/XylS family members is also discussed. PMID- 15784969 TI - Synthesis of lipophilic poly-lauroyl-(+)-catechins and radical-scavenging activity. AB - Lipophilic catechins were synthesized to improve absorption into living bodies and obtain new antioxidants effective in lipid bilayers. The hydroxyl (OH) groups of (+)-catechin was acylated randomly using lauroyl chloride. The mixture was separated by preparative HPLC, and 3-lauroyl-, 3',4'-dilauroyl- and 3,3',4' trilauroyl-catechins (3-LC, 3',4'-LC, and 3,3',4'-LC) were obtained, their structures being determined by (1)H NMR. Their radical scavenging activity was measured in a ethanol solution using the 1,1-diphenyl-2-picrylhydrazyl radical, and was compared with that of (+)-catechin. The activity of 3-LC was almost same as that of (+)-catechin, but those of 3',4'-LC and 3,3',4'-LC were small, showing that the blocking of phenolic OH groups in the B ring lowered the activity. The scavenging activity on lipophilic radicals in a liposome system was also measured, and the activities were in the order of 3-LC > 3',4'-LC = (+)-catechin. These results suggested that radical scavenging activity in the lipid membrane depended not only on the number and the relative positions of phenolic OH groups of catechins but also on affinity to the membrane. PMID- 15784970 TI - Potential ability of hot water adzuki (Vigna angularis) extracts to inhibit the adhesion, invasion, and metastasis of murine B16 melanoma cells. AB - The 40% ethanol eluent of the fraction of hot-water extract from adzuki beans (EtEx.40) adsorbed onto DIAION HP-20 resin has many biological activities, for example, antioxidant, antitumorigenesis, and intestinal alpha-glucosidase suppressing activities. This study examined the inhibitory effect of EtEx.40 on experimental lung metastasis and the invasion of B16-BL6 melanoma cells. EtEx.40 was found significantly to reduce the number of tumor colonies. It also inhibited the adhesion and migration of B16-BL6 melanoma cells into extracellular matrix components and their invasion into reconstituted basement membrane (matrigel) without affecting cell proliferation in vitro. These in vivo data suggest that EtEx.40 possesses a strong antimetastatic ability, which might be a lead compound in functional food development. PMID- 15784971 TI - The N-terminal regions of beta and gamma subunits lower the solubility of adenosylcobalamin-dependent diol dehydratase. AB - Adenosylcobalamin-dependent diol dehydratase is one of essential components of carboxysome-like polyhedral bodies. It exists as a heterohexamer (alphabetagamma)(2), and its activity is recovered in a precipitant fraction of Klebsiella oxytoca and overexpressing Escherichia coli cells. Limited proteolysis of the enzyme with trypsin converted the enzyme into a highly soluble form without loss of enzyme activity. The N-terminal amino acid sequencing of the enzyme thus solubilized indicated that the N-terminal 20 and 16 amino acid residues had been removed from the beta and gamma subunits, respectively. Mutant enzymes with the same N-terminal truncations of either or both of the beta and gamma subunits were expressed on a high level in E. coli cells. All the mutant enzymes obtained were expressed in a soluble, active form. These results indicate that the N-terminal regions of the beta and gamma subunits lower the solubility of diol dehydratase. The mutant enzyme with the N-terminal truncations of both beta and gamma subunits was essentially indistinguishable in catalytic properties from recombinant wild-type enzyme or the enzyme purified from K. oxytoca in a soluble form. PMID- 15784972 TI - Purification and characterization of inducible cephalexin synthesizing enzyme in Gluconobacter oxydans. AB - Cephalexin synthesizing enzyme (CSE) of Gluconobacter oxydans ATCC 9324 was purified up to about 940-fold at a yield of 12%. CSE biosynthesis in G. oxydans was found inducible in the presence of D-phenylglycine but not its substrate phenylglycine methyl ester. The purified enzyme was shown homogeneous on SDS-PAGE and exhibited a specific activity of 440 U per mg protein. The apparent molecular mass of the native enzyme was estimated to be 70 kDa over a Superdex 200 gel filtration column and 68 kDa on SDS-PAGE, indicating that the native enzyme is a monomer. Its isoelectric focusing point is 7.1, indicating a neutral character. The enzyme had maximal activity around pH 6.0 to 6.5, and this activity was thermally stable up to 40 degrees C. Synthesis of cephalexin from D-phenylglycine methyl ester and 7-amino-3-deacetoxycephalosporanic acid (7-ADCA) by the purified CSE was demonstrated. Its L-enantiomer was not accepted by CSE. Apart from cephalexin, ampicillin was also synthesized by the purified CSE from its acyl precursors and 6-aminopenicillanic acid (6-APA). Substrate specificity studies indicated that the enzyme required a free alpha amino group and an activated carboxyl group as a methyl ester of D-form phenylglycine. Interestingly, the purified enzyme did not catalyze hydrolysis of its products, e.g., cephalexin, cephradine, and ampicillin, in contrast to enzymes from other strains of Pseudomonadaceae. PMID- 15784973 TI - Effect of culture conditions on production of 5-aminolevulinic acid by recombinant Escherichia coli. AB - Aminolevulinic acid (ALA) is formed by the enzyme ALA synthase (hemA gene). Then ALA is converted to Porphobilinogen (PBG) by the ALA dehydratase (hemB gene). For the overproduction of ALA, we used an Escherichia coli BL21(DE3) containing a hemA gene from Bradyrhzobium japonicum, which was created in our previous work. The effects of pH on the ALA synthase and ALA dehydratase were investigated. The ALA synthase and ALA dehydratase activities were dependent on the pH of the medium, with maximal activities occurring at pH 6.5 and 8.0 respectively. At pH 6.5, extracellular ALA reached 23 mM in a jar-fermenter. In addition, the effects of some nutritional factors, such as nitrogen source and the ratio of carbon to nitrogen (C/N) on the fermentative production of ALA were investigated. The highest ALA production was found with 8:1 of C/N ratio. Among various nitrogen sources, the tryptone might be a useful one for ALA production. PMID- 15784974 TI - Cloning and heterologous expression of the antibiotic peptide (ABP) genes from Rhizopus oligosporus NBRC 8631. AB - We carried out protein sequencing of purified Antibiotic Peptide (ABP), and cloned two genes encoding this peptide as abp1 and abp2, from Rhizopus oligosporus NBRC 8631. Both genes contain an almost identical 231-bp segment, with only 3 nucleotide substitutions, encoding a 77 amino acid peptide. The abp gene product comprises a 28 amino acid signal sequence and a 49 amino acid mature peptide. Northern blot analysis showed that at least one of the abp genes is transcribed in R. oligosporus NBRC 8631. A truncated form of abp1 encoding only the mature peptide was fused with the alpha-factor signal peptide and engineered for expression in Pichia pastoris SMD1168H. Culture broth of the recombinant Pichia displayed ABP activity against Bacillus subtilis NBRC 3335 after induction of heterologous gene expression. This result indicates that mature ABP formed the active structure without the aid of other factors from R. oligosporus, and was secreted. PMID- 15784975 TI - Lipid peroxidation of a human hepatoma cell line (HepG2) after incorporation of linoleic acid, arachidonic acid, and docosahexaenoic acid. AB - Lipid peroxidation of human heptoma cell line, HepG2, after incorporation of linoleic acid (LA), arachidonic acid (AA), and docosahexaenoic acid (DHA) was measured with a fluorescent probe and gas chromatography-mass spectrometry (GC MS) analysis. The analysis with a fluorescent probe showed that incorporation of each polyunsaturated fatty acid (PUFA) enhanced the cellular lipid peroxidation level, but there was little difference in the effect of LA, AA, or DHA on the enhancement of cellular lipid peroxidation. The fluorescent analysis also showed that the addition of H(2)O(2) (0.5 mM) enhanced the cellular lipid peroxidation levels in LA and AA supplemented cells as compared with those without H(2)O(2). However, the enhancement of lipid peroxidation by H(2)O(2) was not observed in DHA-supplemented cells. The same result was obtained in the GC-MS analysis of total amounts of monohydroperoxides (MHP) formed in the cellular phospholipid oxidation. In this case, the main source for MHP was LA in LA-, AA-, and DHA supplemented cells. A significant amount of AA-MHP and a small amount of DHA-MHP were observed in AA- and DHA-supplemented cells respectively. GC-MS analysis also indicated the specific positional distribution of DHA-MHP isomers. The isomers were formed only by hydrogen abstraction at the C-18 (16-MHP + 20-MHP; 46.5%), C 6 (4-MHP + 8-MHP; 38.5%), and C-12 (10-MHP + 14-MHP; 15.1%) positions, but not at the C-9 or C-15 positions. PMID- 15784976 TI - A series of crystal structures of a meta-cleavage product hydrolase from Pseudomonas fluorescens IP01 (CumD) complexed with various cleavage products. AB - Meta-cleavage product hydrolase (MCP-hydrolase) is one of the key enzymes in the microbial degradation of aromatic compounds. MCP-hydrolase produces 2 hydroxypenta-2,4-dienoate and various organic acids, according to the C6 substituent of the substrate. Comprehensive analysis of the substrate specificity of the MCP-hydrolase from Pseudomonas fluorescens IP01 (CumD) was carried out by determining the kinetic parameters for nine substrates and crystal structures complexed with eight cleavage products. CumD preferred substrates with long non branched C6 substituents, but did not effectively hydrolyze a substrate with a phenyl group. Superimposition of the complex structures indicated that benzoate was bound in a significantly different direction than other aliphatic cleavage products. The directions of the bound organic acids appeared to be related with the k(cat) values of the corresponding substrates. The Ile139 and Trp143 residues on helix alpha4 appeared to cause steric hindrance with the aromatic ring of the substrate, which hampers base-catalyzed attack by water. PMID- 15784977 TI - The effect of protozoa on the composition of rumen bacteria in cattle using 16S rRNA gene clone libraries. AB - The effect of the presence of protozoa on the composition of rumen bacteria was investigated in cattle. Seven castrated Holstein cattle were divided into two groups: four faunated and three unfaunated, and 16S ribosomal RNA gene (rDNA) clonal libraries were constructed. A total of 312 clones were sequenced across 1,500 bp. The 151 sequences of the faunated cattle were classified into 98 operational taxonomic units (OTUs) having at least 97% similarity. The sequences derived from the faunated cattle were classified into Firmicutes (59.7%), Bacteroidetes (34.4%), Spirochaetes (2.6%), Actinobacteria (2.0%), and Proteobacteria (1.3%). Bacteroides and Prevotella (34.4%) were the major groups in the faunated cattle. The 161 sequences in the unfaunated cattle were classified into 72 OTUs. The sequences derived from the unfaunated libraries were classified into Firmicutes (65.7%), Bacteroidetes (31.1%), Proteobacteria (1.9%), and Spirochaetes (1.2%). The Clostridium botulinum group and its relatives (36.0%) were the major groups in the unfaunated cattle. An analysis by the computer program LIBSHUFF clarified that the presence of ruminal protozoa markedly affected the composition of rumen bacteria. PMID- 15784978 TI - Superoxide dismutase from the silkworm, Bombyx mori: sequence, distribution, and overexpression. AB - Superoxide dismutase (SOD) is an enzyme facilitating the removal of superoxide anions from living organisms. This study focused on SOD from the silkworm, Bombyx mori (bmSOD). cDNA encoding bmSOD was amplified by reverse transcriptase polymerase chain reaction. The deduced amino acid sequence of bmSOD indicated that the residues forming the Cu/Zn binding site are conserved and that the sequence is in 60% identity to that of the Drosophila melanogaster. B. mori SOD was also close to the D. melanogaster SOD in a phylogenetic tree. The bmSOD mRNA and the enzyme activity were widely distributed in diverse tissues. bmSOD functionally overexpressed in Escherichia coli in a soluble form was purified, and its stability was examined. bmSOD at 4 degrees C retained almost all of its original activity after incubation at pH 4-11 for 24 h. Incubation (pH 7) for 30 min at temperatures below 40 degrees C also affected activity insignificantly. PMID- 15784979 TI - FR207944, an antifungal antibiotic from Chaetomium sp. no. 217 I. Taxonomy, fermentation, and biological properties. AB - An antifungal antibiotic, FR207944, was isolated from the culture broth of a fungal strain Chaetomium sp. no. 217. FR207944 is a triterpene glucoside with antifungal activity against Aspergillus fumigatus and Candida albicans. Specifically, FR207944 exhibits in vitro and in vivo antifungal activity against A. fumigatus. The effects of FR207944 on the morphology of A. fumigatus were shown to be similar to those of FR901379, a known 1,3-beta-glucan synthase inhibitor. The MECs of FR207944 against A. fumigatus FP1305 and C. albicans FP633 in micro-broth dilution test were 0.039 and 1.6 mug/ml respectively. FR207944 showed good potency by subcutaneous injection and oral administration against A. fumigatus in a murine systemic infection model, with ED(50)s of 5.7 and 17 mg/kg respectively. PMID- 15784980 TI - Purification and characterization of a novel alpha-glucuronidase from Aspergillus niger specific for O-alpha-D-glucosyluronic acid alpha-D-glucosiduronic acid. AB - A new alpha-glucuronidase that specifically hydrolyzed O-alpha-D-glucosyluronic acid alpha-D-glucosiduronic acid (trehalose dicarboxylate, TreDC) was purified from a commercial enzyme preparation from Aspergillus niger, and its properties were examined. The enzyme did not degrade O-alpha-D-glucosyluronic acid alpha-D glucoside, O-alpha-D-glucosyluronic acid beta-D-glucosiduronic acid, O-alpha-D glucosyluronic acid-(1-->2)-beta-D-fructosiduronic acid, p-nitrophenyl-O-alpha-D glucosiduronic acid, methyl-O-alpha-D-glucosiduronic acid, or 6-O-alpha-(4-O alpha-D-glucosyluronic acid)-D-glucosyl-beta-cyclodextrine. Furthermore, it showed no activity on alpha-glucuronyl linkages of 4-O-methyl-D-glucosyluronic acid-alpha-(1-->2)-xylooligosaccharides, derived from xylan, a supposed substrate of alpha-glucuronidases.The molecular mass of the enzyme was estimated to be 120 kDa by gel filtration and 58 kDa by SDS-PAGE suggesting, the enzyme is composed of two identical subunits. It was most active at pH 3.0-3.5 and at 40 degrees C. It was stable in pH 2.0-4.5 and below 30 degrees C. It hydrolyzed O-alpha-D glucosyluronic acid alpha-D-glucosiduronic acid to produce alpha- and beta anomers of D-glucuronic acid in an equimolar ratio. This result suggests that inversion of the anomeric configuration of the substrate is involved in the hydrolysis mechanism. PMID- 15784981 TI - Purification and characterization of hydantoin racemase from Microbacterium liquefaciens AJ 3912. AB - A hydantoin racemase that catalyzed the racemization of 5-benzyl-hydantoin was detected in a cell-free extract of Microbacterium liquefaciens AJ 3912, a bacterial strain known to produce L-amino acids from their corresponding DL-5 substituted-hydantoins. This hydantoin racemase was purified 658-fold to electrophoretic homogeneity by serial chromatography. The N-terminal amino acid sequence of the enzyme showed homology with known hydantoin racemases from other microorganisms. The apparent molecular mass of the purified enzyme was 27 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and 117 kDa on gel-filtration in the purification conditions, indicating a homotetrameric structure. The purified enzyme exhibited optimal activity at pH 8.2 and 55 degrees C, and showed a chiral preference for L-5-benzyl- rather than D-5-benzyl hydantoin. PMID- 15784982 TI - Static suppression of tomato bacterial wilt by bacterial coagulation using a new functional polymer that coagulates bacterial cells and is highly biodegradable. AB - Tomato bacterial wilt by Ralstonia solanacearum was suppressed by coagulation of bacterial cells without disinfection using a copolymer of methyl methacrylate with N-benzyl-4-vinylpyridinium chloride in a molar ratio of 3:1 (PMMA-co-BVP) as a polymeric coagulant for bacterial cells. When 10 mg/kg of PMMA-co-BVP was added to soil before transplanting of tomato seedlings, and 2 mg/kg was supplemented once a week after transplanting, a 51% reduction of appearance and a 54% reduction of index of symptoms were observed. PMMA-co-BVP did not exhibit bactericidal activity against R. solanacearum, and coagulation of the bacterial cells appeared to reduce the opportunity for infectious contact of roots of tomato with cells of R. solanacearum, and resulted in disease suppression. PMMA co-BVP was shown to be highly biodegradable, and the half-life was 5.1 d when treated with activated sludge in soil. PMID- 15784983 TI - Highly active mutants of carbonyl reductase S1 with inverted coenzyme specificity and production of optically active alcohols. AB - A wild type NADPH-dependent carbonyl reductase from Candida magnoliae (reductase S1) has been found not to utilize NADH as a coenzyme. A mutation to exchange the coenzyme specificity in reductase S1 has been designed by computer-aided methods, including three-dimensional structure modeling and in silico screening of enzyme mutants. Site-directed mutagenesis has been used to introduce systematic substitutions of seven or eight amino acid residues onto the adenosine-binding pocket of the enzyme according to rational computational design. The resulting S1 mutants show NADH-dependency and have lost their ability to utilize NADPH as a coenzyme, but retain those catalytic activities. Kinetic parameter V(max) and K(m) values of those mutants for NADH are 1/3- to 1/10-fold those of the wild type enzyme for NADPH. As a model system for industrial production of optically active alcohols, the S1 mutants can be applied to an asymmetric reduction of ketones, cooperating with a coenzyme-regeneration system that uses an NAD dependent formate dehydrogenase. PMID- 15784984 TI - Structure of beta-glucan oligomer from laminarin and its effect on human monocytes to inhibit the proliferation of U937 cells. AB - We analyzed the human monocyte-stimulating ability of laminarin from Eisenia bicyclis, lichenan from Cetraria islandica, and their oligomers depolymerized with endo-1,3-beta-glucanase from Arthrobacter sp. The respective beta-glucan oligomers with different degrees of polymerization (DP) were fractionated from hydrolytic products of laminarin and lichenan using gel-filtration chromatography. The monocyte-conditioned medium pre-cultured in the presence of a fraction of beta-glucan oligomer (DP>/=8) from laminarin exhibited inhibitory activity against the proliferation of human myeloid leukemia U937 cells, while those pre-cultured with other beta-glucan oligomers and the original laminarin and lichenan showed little or no activity. NMR analysis indicated that the beta glucan oligomer (DP>/=8) has an average DP value of 13, and its ratio of beta-1,3 to beta-1,6-linkages in glucopyranose units was estimated to be 1.3:1. These results indicate that the beta-1,3-glucan oligomer with a higher content of beta 1,6-linkage stimulates monocytes to inhibit the proliferation of U937 cells. PMID- 15784985 TI - Effect of bread containing resistant starch on postprandial blood glucose levels in humans. AB - We examined the inhibitory effect of a single ingestion of bread containing resistant starch (bread containing about 6 g of resistant starch derived from tapioca per 2 slices) (test food) on the postprandial increase in blood glucose in male and female adults with a fasting blood glucose level between 100 and 140 mg/dl. Bread not containing resistant starch (placebo) was used as the control. The study was conducted in 20 subjects (9 men and 11 women with a mean age of 50.5+/-7.5 years) using the crossover method, with a single ingestion of either bread containing resistant starch or the placebo. Blood glucose and insulin were measured before ingestion, and at 0.5, 1, 1.5, and 2 h after ingestion. The blood glucose level before ingestion was stratified into a borderline group (blood glucose level >/= 111 mg/dl) and a normal group (blood glucose level bl), was 0.13 nmol/min/mg of protein. This permeation rate is nearly the same as that of 5 mM chlorogenic acid (CLA) and gallic acid, which are paracellularly transported compounds. Almost all of the apically loaded RA was retained on the apical side, and J(ap-->bl) was inversely correlated with paracellular permeability. These results indicate that RA transport was mainly via paracelluar diffusion, and the intestinal absorption efficiency of RA was low. Furthermore, RA appeared to be unsusceptible to hydrolysis by mucosa esterase in Caco-2 cells. These results, together with our previous work (J. Agric. Food Chem., 52, 2518-2526 (2004), J. Agric. Food Chem., 52, 6418-6424 (2004)) suggest that the majority of RA is further metabolized and degraded into m-coumaric and hydroxylated phenylpropionic acids by gut microflora, which are then efficiently absorbed and distributed by the monocarboxylic acid transporter (MCT) within the body. The potential of orally administered RA in vivo will be further investigated. PMID- 15784989 TI - Functional analysis of two solanesyl diphosphate synthases from Arabidopsis thaliana. AB - Solanesyl diphosphate (SPP) is regarded as the precursor of the side-chains of both plastoquinone and ubiquinone in Arabidopsis thaliana. We previously analyzed A. thaliana SPP synthase (At-SPS1) (Hirooka et al., Biochem. J., 370, 679-686 (2003)). In this study, we cloned a second SPP synthase (At-SPS2) gene from A. thaliana and characterized the recombinant protein. Kinetic analysis indicated that At-SPS2 prefers geranylgeranyl diphosphate to farnesyl diphosphate as the allylic substrate. Several of its features, including the substrate preference, were similar to those of At-SPS1. These data indicate that At-SPS1 and At-SPS2 share their basic catalytic machinery. Moreover, analysis of the subcellular localization by the transient expression of green fluorescent protein-fusion proteins showed that At-SPS2 is transported into chloroplasts, whereas At-SPS1 is likely to be localized in the endoplasmic reticulum in the A. thaliana cells. It is known that the ubiquinone side-chain originates from isopentenyl diphosphate derived from the cytosolic mevalonate pathway, while the plastoquinone side-chain is synthesized from isopentenyl diphosphate derived from the plastidial methylerythritol phosphate pathway. Based on this information, we propose that At SPS1 contributes to the biosynthesis of the ubiquinone side-chain and that At SPS2 supplies the precursor of the plastoquinone side-chain in A. thaliana. PMID- 15784990 TI - Cloning and expression of a Bacillus circulans KA-304 gene encoding chitinase I, which participates in protoplast formation of Schizophyllum commune. AB - KA-prep, a culture filtrate of Bacillus circulans KA-304 grown on a cell-wall preparation of Schizophyllum commune, has an activity to form protoplasts from S. commune mycelia, and a combination of alpha-1,3-glucanase and chitinase I, isolated from KA-prep, brings about the protoplast-forming activity. The gene of chitinase I was cloned from B. circulans KA-304 into pGEM-T Easy vector. The gene consists of 1,239 nucleotides, which encodes 413 amino acids including a putative signal peptide (24 amino acid residues). The molecular weight of 40,510, calculated depending on the open reading frame without the putative signal peptide, coincided with the apparent molecular weight of 41,000 of purified chitinase I estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The C-terminal domain of the deduced amino acid sequence showed high similarity to that of family 19 chitinases of actinomycetes and other organisms, indicating that chitinase I is the first example of family 19 chitinase in Bacillus species. Recombinant chitinase I without the putative signal peptide was expressed in Escherichia coli Rosetta-gami B (DE 3). The properties of the purified recombinant enzyme were almost the same as those of chitinase I purified from KA-prep, and showed the protoplast-forming activity when it was combined with alpha-1,3-glucanase from KA-prep. Recombinant chitinase I as well as the native enzyme inhibited hyphal extension of Trichoderma reesei. PMID- 15784991 TI - Preparation of functional single-chain antibodies against bioactive gibberellins by utilizing randomly mutagenized phage-display libraries. AB - Screening randomly mutagenized proteins displayed on a phage surface by biopanning is a powerful strategy to obtain evolved clones with improved properties such as higher stability and functionality. We utilized this method to overcome the problem that functional single-chain antibodies against active gibberellins, a class of plant hormones, can not be prepared by some of the conventional methods. Single-chain antibody libraries with random mutations were constructed from two independent anti-bioactive gibberellin monoclonal antibody lines in a phagemid vector, so that the mutagenized scFvs were expressed in a phage-displayed form upon helper phage infection. From both libraries, scFv clones with binding activity to GA(4) were successfully obtained by successive rounds of biopanning against BSA-GA(4), the original immunogen. The results are highly suggestive that this approach might be a general solution when a single chain antibody does not show binding activity. We found further that a ribosomal frameshift to complement a nonsense mutation frequently occurred in an amber suppressor strain of E. coli TG1, resulting in the display of a functional antibody, while such a nonsense mutant failed to produce a soluble antibody in a non-amber suppressor strain. This result explains at least partly why single chain antibodies are sometimes functional only in a phage-displayed form, not in a soluble form. PMID- 15784992 TI - Protective effect of epigallocatechin gallate and esculetin on oxidative DNA damage induced by psoralen plus ultraviolet-A therapy. AB - We examined antioxidants exhibiting no effects on DNA cross-linking, which is the basis of psoralen and ultraviolet-A therapy for skin diseases, and suppressing oxidative DNA damage incidental to the therapy. Epigallocatechin gallate and esculetin effectively suppressed oxidative DNA damage with little effect on the formation of DNA cross-linking. These antioxidants might be useful in suppressing the adverse reaction induced by this therapy. PMID- 15784993 TI - Effects of serum deprivation on expression of proteolytic-related genes in chick myotube cultures. AB - We previously reported that serum deprivation stimulates myofibrillar proteolysis in chick myotubes. In the present study, we examined the effect of serum deprivation on expression of the proteolytic-related genes (ubiquitin, proteasome, calpains, and cathepsin B) by real-time PCR of cDNA in chick myotubes. Myotubes were incubated with serum-free medium for 24 h. Ubiquitin and proteasome subunits (C1 and C2) and calpains (m-, mu-, and p94/calpain-3) but not cathepsin B mRNA expression were increased by serum deprivation. These results indicate that serum deprivation stimulates ubiquitin-proteasome and calpain proteolytic pathways, resulting in an increase in myofibrillar proteolysis in chick myotubes. PMID- 15784994 TI - Biosynthesis of the marine antibiotic pentabromopseudilin. 2. The pyrrole ring. AB - The biosynthesis of the potent marine antibiotic, pentabromopseudilin (1), was investigated. Feeding studies with Alteromonas luteoviolaceus were performed on a defined medium. D,L-[5-(13)C]proline was incorporated symmetrically, demonstrating that the pyrrole ring of pentabromopseudilin is derived from proline. PMID- 15784995 TI - Effect of molecular mass on antitumor activity of heteropolysaccharide from Poria cocos. AB - A water-soluble heteropolysaccharide ac-PCM0 from Poria cocos was successfully fractionated using a preparative size exclusion chromatography (SEC) column, and their weight-average molecular mass (M(w)) was characterized by analytical SEC combined with laser light scattering (SEC-LLS). The results indicate that the fractions having relatively high M(w) exhibited higher inhibition ratio in vivo antitumor activity than those having M(w) below 3.29 x 10(4). However, the relatively low molecular mass was beneficial to the in vitro antitumor activity. Moreover, ac-PCM0 has a significantly higher enhancement ratio of the body weight than 5-fluorouracil, and its 50% lethal dose is above 1250 mg/kg, indicating a nontoxic nature. PMID- 15784996 TI - Production of heterogeneous dimer lignostilbenedioxygenase II from lsdA and lsdB in Escherichia coli cells. AB - The lignostilbenedioxygenase isozyme I and III genes, lsdA and lsdB, were coexpressed within the same Escherichia coli cells. The lignostilbenedioxygenase isozymes produced were separated on QAE anion-exchange column chromatography. Three parts of active fractions corresponding to alphaalpha, alphabeta, and betabeta dimers were detected. The purified isozyme from second active fractions corresponded to the native heterogeneous isozyme II. PMID- 15784997 TI - Molecular monitoring of the developmental bacterial community in the gastrointestinal tract of Japanese infants. AB - The dynamics of the developmental bacterial community in the Japanese neonatal gastrointestinal tract were examined by monitoring 16S ribosomal RNA gene (rDNA) diversity in fecal samples by PCR and denaturing gradient gel electrophoresis (DGGE). The results showed a certain pattern common in infants without antibiotic treatment, in which aerobes, e.g., Pseudomonas, appeared first and were then immediately replaced by facultative anaerobe, Enterococcus, Streptococcus, and Enterobacteriaceae through the first month, and finally strictly anaerobic Bifidobactrerium appeared. PMID- 15784999 TI - Microtubule-cyclodextrin conjugate: functionalization of motile filament with molecular inclusion ability. AB - A microtubule-beta-cyclodextrin conjugate was prepared on a kinesin-adsorbed glass surface by chemical and biochemical means. Fluorescence microscope observation and a motility assay indicated that the conjugate simultaneously expressed an inherent motor function and an inclusion property. PMID- 15784998 TI - Amino acid sequence and antimicrobial activity of chitin-binding peptides, Pp-AMP 1 and Pp-AMP 2, from Japanese bamboo shoots (Phyllostachys pubescens). AB - Two novel chitin-binding peptides, designated Pp-AMP 1 and Pp-AMP 2, which had antimicrobial activity against pathogenic bacteria and fungi, were purified from Japanese bamboo shoots (Phyllostachys pubescens) by a simple procedure based on chitin affinity chromatography. They had the common structural features of the plant defensin family, but they could not be grouped in any type of that family. They showed a high degree of homology to mistletoe toxins. PMID- 15785000 TI - Cloning and sequence analysis of the micronuclear and macronuclear gene encoding Rab protein of Euplotes octocarinatus. AB - The DNA in a micronucleus undergoes remarkable rearrangements when it develops into a macronucleus after cell mating in the hypotrichous ciliate. A Rab gene was isolated from the macronuclear plasmid mini-library of Euplotes octocarinatus. A micronuclear version of the Rab gene was amplified by polymerase chain reaction (PCR). The macronuclear DNA molecule carrying the Rab gene is 767 bp long and shows characteristics typical of macronuclear chromosomes of hypotrichous ciliates. Three of the five cysteines are encoded by the opal codon UGA. The deduced protein is a 207-amino acid (aa) with a molecular mass of 23 kDa. The protein shares 36% identity with Rab 1 protein of Plasmodium and yeast. Analysis of the sequences indicated that the micronuclear version of the Rab gene contains two internal eliminated sequences, internal eliminated sequence (IES)1 and IES2. IES1 is flanked by a pair of hepta-nucleotide 5'-AAATTTT-3' direct repeats, and IES2 is flanked by 5'-TA-3' direct repeats. PMID- 15785001 TI - Anti-invasive activity of niacin and trigonelline against cancer cells. AB - The effects of niacin, namely, nicotinic acid and nicotinamide, and trigonelline on the proliferation and invasion of cancer cells were studied using a rat ascites hepatoma cell line of AH109A in culture. Niacin and trigonelline inhibited the invasion of hepatoma cells at concentrations of 2.5-40 microM without affecting proliferation. Hepatoma cells previously cultured with a reactive oxygen species (ROS)-generating system showed increased invasive activity. Niacin and trigonelline suppressed this ROS-potentiated invasive capacity through simultaneous treatment of AH109A cells with the ROS-generating system. The present study indicates for the first time the anti-invasive activities of niacin and trigonelline against cancer cells. PMID- 15785002 TI - Microbial production of L-ascorbic acid from D-sorbitol, L-sorbose, L-gulose, and L-sorbosone by Ketogulonicigenium vulgare DSM 4025. AB - Ketogulonicigenium vulgare DSM 4025, known as a 2-keto-L-gulonic acid producing strain from L-sorbose via L-sorbosone, surprisingly produced L-ascorbic acid from D-sorbitol, L-sorbose, L-gulose, and L-sorbosone as the substrate under a growing or resting condition. As the best result, K. vulgare DSM 4025 produced 1.37 g per liter of L-AA from 5.00 g per liter of L-sorbosone during 4 h incubation time at 30 degrees C under the resting cell condition having 5.70 g per liter of wet cells. The precursor of L-AA formation from D-sorbitol and L-sorbose, except for L-gulose, was thought to be the putative furanose form of L-sorbosone. This is the first time it is reported that bacteria can produce vitamin C via L sorbosone. PMID- 15785003 TI - Association of genetic polymorphisms of sodium-calcium exchanger 1 gene, NCX1, with hypertension in a Japanese general population. AB - The Na+/Ca2+ exchanger (NCX) is a membrane protein involved in calcium homeostasis, catalyzing the exchange of one Ca2+ ion for three Na+ ions across the cell membrane. The Na+/Ca2+ exchange has been suggested to play a role in the pathogenesis of hypertension. Therefore, we examined whether genetic variations in NCX1 were associated with hypertension. Among 15 polymorphisms identified in 96 hypertensive subjects by sequencing the entire exon and promoter regions of NCX1, 7 representative polymorphisms with a minor allele frequency of greater than 4% were genotyped in 1,865 individuals, of whom 787 were hypertensive and 1,072 were normotensive. These subjects were residents of Suita City and were randomly selected as a population for the Suita cohort study. Multivariate logistic regression analysis performed after adjusting for age, body mass index, hyperlipidemia, diabetes mellitus, smoking, and drinking revealed that the 23200T>C and -23181T>C polymorphisms in the 5' upstream region of exon 1c were significantly associated with hypertension in men (-23200T>C: CC vs. TC+TT: odds ratio=0.61; 95% confidence intervals: 0.39 to 0.97; p =0.04) and in women ( 23181T>C: CC vs. TC+TT: odds ratio=1.45; 95% confidence intervals: 1.04 to 2.02; p =0.03), respectively. Thus, our study suggests that NCX1 is one of the genes related to susceptibility to essential hypertension in the Japanese general population. PMID- 15785004 TI - Trends in prevalence, awareness, treatment and control of hypertension in the middle-aged population of China, 1992-1998. AB - The objective of this study was to analyze the trends in prevalence, awareness, treatment and control of hypertension in the middle-aged population of China. There were about 1,000 participants each from 13 different study populations that were examined in surveys conducted from 1992-1994 (n=18,746) and in 1998 (n=13,504) in conjunction with the China Multi-Center Study of Cardiovascular Epidemiology. Half the subjects were men and half were women; their ages ranged from 35 to 59 years. Hypertension was defined as systolic blood pressure > or =140 mmHg, diastolic blood pressure > or =90 mmHg, and/or current treatment with antihypertensive medications. Hypertension awareness and treatment were assessed with a standardized questionnaire. Hypertension control was defined as blood pressure measurements of less than 140/90 mmHg. The results showed that 24.0% of participants had hypertension in 1998, an increase of 2.3% from 1992-1994 (p <0.05). The prevalence of hypertension was 25.4% higher in urban than in rural areas, and was higher in men than in women. Among hypertensives, 42.6% were aware of their hypertension (a 5.3% increase compared with 1992-1994, p <0.05), 31.1% were treated (a 3.8% increase, p <0.05), and 6.0% were controlled (a 2.6% increase, p <0.05). The rates of awareness, treatment and control were higher in women than men, and higher in urban than rural areas. For treated hypertensives, the rate of control increased from 12.7% in 1992-1994 to 19.9% in 1998 (p <0.05). These findings indicate that hypertension prevalence is increasing in China. Control rates, while improving, still remain low. This implies that effective public health measures are needed to enhance the awareness, treatment, and control rates in the Chinese population. PMID- 15785006 TI - Plasma mitochondrial coupling factor 6 in patients with acute myocardial infarction. AB - Previous studies have shown that mitochondrial coupling factor 6 (CF6) is an endogenous peptide that inhibits prostacyclin (PGI2) synthesis in vascular endothelial cells. In this study, we measured the plasma CF6 level of patients with acute myocardial infarction (AMI) to observe dynamic changes of CF6. All patients showed elevated plasma CF6 levels upon admission for treatment of AMI. Their CF6 levels peaked approximately 72 h after the onset of AMI and remained high for 7 days. At 7 days, their CF6 levels decreased to the level seen upon admission, but not to within a normal range. Hyperlipidemic patients had significantly greater CF6 levels at 24 h after onset of AMI than patients with a normal lipid profile. On admission, the plasma CF6 level in patients with a cardiac function of Killip class > or =II was higher than that in patients with a Killip class I cardiac function. At 3 days after the onset of AMI, the plasma CF6 levels of patients with a creatinine kinase (CK) peak value > or =1,500 units/l were significantly higher than those of patients with a CK peak value <1,500 units/l (p =0.05). At 7 days after the onset of AMI, the plasma CF6 levels of patients who received no reperfusion were significantly higher than those of patients who received a successful reperfusion. The plasma CF6 levels of AMI patients at admission, at 24 h, and at 3 days after onset of symptoms correlated positively with the cardiac function by Killip classification, respectively. At 24 h after onset of AMI, the plasma CF6 levels correlated positively with plasma total cholesterol levels and low-density lipoprotein levels. At 3 days, the plasma level of CF6 correlated positively with the plasma CK peak value and correlated negatively with left ventricular ejection fraction. These results suggest that the plasma CF6 level was elevated in patients with AMI. PMID- 15785005 TI - Variable number of tandem repeat of the 5'-flanking region of type-C human natriuretic peptide receptor gene influences blood pressure levels in obesity associated hypertension. AB - The gene for natriuretic peptides receptor C (NPRC), which is a candidate susceptibility gene for essential hypertension (EH) or obese hypertension, plays a key role in the regulation of plasma levels and biological effects of natriuretic peptides. The aims of the present study were to find new genetic markers in the 5'-flanking region of the NPRC gene and to assess relationships between variants and phenotypes of EH, including EH in obese patients. Using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis, we discovered a novel six-nucleotide repeat polymorphism located 4 base pairs (bp) upstream of the major transcriptional initiation site. We performed association analysis of this variable number of tandem repeat (VNTR) in 242 EH patients and 212 normotensive controls (NT). Although no significant difference in overall frequency of VNTR was found between NT and EH groups, the blood pressure level of EH patients with the 5/6 genotype was significantly higher in obese subjects. This suggests that the VNTR of the 5'-flanking region of the NPRC gene influences blood pressure levels in obesity-associated hypertension. PMID- 15785007 TI - Cardiovascular risk factor profiles and endothelial function in coronary artery disease patients treated with statins. AB - Although endothelial dysfunction is associated with cardiovascular risk factors and is improved by cholesterol-lowering therapy, the relationship between endothelial function and cardiovascular risk factor profiles has not been fully investigated in coronary artery disease patients who have been treated with statins. We investigated endothelial function in male hypercholesterolemic patients (n=53) who underwent statin therapy over 6 months in a cross-sectional study. Patients were classified into three groups based on the results of coronary angiography: a normal coronary artery group (n=15), an angina pectoris group (n=20) and a myocardial infarction group (n=18). Endothelial function was assessed by measuring flow-mediated dilatation after reactive hyperemia in the brachial artery, and serum lipid, lipoprotein (a), glucose and insulin levels were measured. Significant associations were observed between the status of coronary disease and systolic blood pressure, lipoprotein (a), glucose and insulin levels (p <0.05, respectively), and the levels of these risk factors in the myocardial infarction group were higher than those in the other groups. Flow mediated dilatation was also associated with the status of coronary disease (p <0.05), and the myocardial infarction group showed the lowest levels of flow mediated dilatation (p <0.05). Flow-mediated dilatation was negatively correlated with systolic and diastolic blood pressures, serum levels of lipoprotein (a), glucose and insulin, and the status of coronary disease. Stepwise multiple regression analysis also revealed that lipoprotein (a), diastolic blood pressure and the status of myocardial infarction were significantly correlated with impaired vasodilatation. Serum lipids, age and smoking habit were independent of flow-mediated dilatation. In conclusion, even after cholesterol-lowering treatment, male patients with myocardial infarction still had endothelial dysfunction, and higher levels of lipoprotein (a) may be associated with endothelial dysfunction in such patients. PMID- 15785008 TI - Green coffee bean extract improves human vasoreactivity. AB - Our previous study revealed the antihypertensive effects of green coffee bean extract (GCE) ingestion in spontaneously hypertensive rats. We suggested that this antihypertensive action was due to the fact that GCE contains chlorogenic acid (CQA) as a major phenolic compound, and CQA in turn contains ferulic acid as a metabolic component that acts on nitric oxide (NO) derived from the vascular endothelium. In this study, the effects of GCE on blood vessels were evaluated in healthy males. The subjects were 20 healthy males with reduced vasodilation responses measured by strain gauge plethysmograms (SPG) to ischemic reactive hyperemia. Of the 20 subjects, 10 (mean age, 37.2 years) ingested a test drink containing GCE (CQA: 140 mg/day), and the other 10 (mean age, 34.8 years) ingested a placebo drink for 4 months. During the ingestion period, SPG, pulse wave velocity (PWV), and serum biochemical parameters were measured, and acceleration plethysmograms (APG) were taken. The reactive hyperemia ratio (RHR) in the test drink group began to increase after ingestion for 1 month and was significantly higher (p <0.05) than that in the placebo group after ingestion for 3 months and 4 months. In addition, after ingestion for 4 months, the test drink group showed a significant decrease (p <0.01) in the plasma total homocysteine level compared with the pre-ingestion level. However, there were no significant differences in PWV or APG between the test drink group and the placebo drink group. The improvement in RHR after ingestion of a drink containing GCE suggested an improvement in vasoreactivity by this component. PMID- 15785009 TI - White-coat hypertension contributes to the presence of carotid arteriosclerosis. AB - It remains unclear whether white-coat hypertension is associated with vascular organ damage (e.g., carotid arteriosclerosis) in the same way sustained hypertension is. We therefore compared the progression of carotid arteriosclerosis among Japanese individuals showing normal blood pressures, sustained hypertension or white-coat hypertension. A total of 30 subjects (mean age, 58 years) with white-coat hypertension, 30 (mean age, 54 years) with untreated sustained hypertension who had no plaque formation in the carotid arteries, and 30 normotensive subjects (mean age, 58 years) were enrolled in this study. The white-coat and sustained hypertensive subjects were matched with respect to their clinical blood pressures, but their ambulatory blood pressures differed. Conversely, white-coat hypertensive and normotensive subjects were matched with respect to ambulatory blood pressures, but their clinical blood pressures differed. Carotid intimal-medial thickness was measured by B-mode ultrasonography, and the cross-sectional area of the common carotid artery was calculated. The three groups were similar with respect to age, sex ratio, height, laboratory data and the incidence of smoking. Body weights and body mass indexes were significantly higher among patients with sustained hypertension than among either normotensive or white-coat hypertensive patients. Intimal-medial thicknesses and carotid cross-sectional areas were similar in patients with white coat and sustained hypertension and significantly higher than in normotensive subjects. Collectively, these findings suggest that white-coat hypertension contributed to the presence of carotid arteriosclerosis in our subjects in a manner similar to sustained hypertension. Thus, clinical evaluation of white-coat hypertension should be conducted with the potential for target organ damage in mind. PMID- 15785010 TI - Smoking is associated with silent cerebrovascular disease in a high-risk Japanese community-dwelling population. AB - We aimed to investigate the relationships between smoking and silent cerebrovascular damage. We performed brain MRI to evaluate silent cerebral infarct (SCI) and periventricular hyperintensity (PVH), and carotid ultrasonography to investigate carotid atherosclerotic plaque in 170 high-risk community-dwelling subjects (mean age: 67.2 years; men: 28.7%) who met more than 3 of the following 9 criteria: 1) high blood pressure (BP); 2) hypercholesterolemia; 3) left ventricular hypertrophy; 4) high hemoglobin A1c; 5) proteinuria; 6) high waist-to-hip ratio; 7) smoking > or =30 cigarettes/day; 8) heavy alcohol intake; 9) family history of stroke. The subjects with SCI (SCI group) were older (70 years vs. 66 years, p=0.004) and had higher systolic BP (SBP) (160 vs. 148 mmHg, p <0.001) and higher carotid plaque score (2.3 vs. 1.5/person, p <0.05) than those without SCI. Among the variables, smoking status (r =0.34, p <0.001), SBP (r =0.28, p <0.001), male gender (r =0.29, p <0.001), left ventricular mass index (r =0.25, p =0.001), and serum creatinine (r =0.20, p =0.006) were significantly correlated with the number of SCIs. Among smokers, the number of SCIs was significantly higher in current smokers than in past smokers (1.9+/-2.2 vs. 0.5+/-0.8, p <0.01). In multiple regression analysis, smoking status (beta =0.183, p =0.045) and SBP (beta =0.196, p =0.011) were independent determinants of the increased number of SCIs. In conclusion, smoking status was an independent determinant of multiple SCIs in a high-risk Japanese community dwelling population. PMID- 15785011 TI - Control of blood pressure as measured at home and office, and comparison with physicians' assessment of control among treated hypertensive patients in Japan: First Report of the Japan Home versus Office Blood Pressure Measurement Evaluation (J-HOME) study. AB - The Japan Home versus Office Blood Pressure Measurement Evaluation (J-HOME) study was conducted to measure the control of blood pressure (BP) as evaluated by home BP measurement among 3,400 patients with essential hypertension (mean age: 66 years; females: 55%) receiving antihypertensive treatment in primary care settings in Japan. The purpose of this first report was to compare characteristics of BP control as measured at home and in the clinic (office) and define their association with BP control as evaluated by physicians. Mean systolic/diastolic BP (SBP/DBP) values were 140/82 mmHg for home BP and 143/81 mmHg for office BP. BP levels were not adequately controlled among approximately 60% of the patients, according to reference values described in the national guidelines (office BP: <140/90 mmHg; home BP: <135/85 mmHg). Even among patients evaluated by physicians as having excellent or fairly good BP control, office and home SBP values were insufficiently controlled in approximately 50%. Although the tendency was more remarkable among older patients, whose recommended target BP levels are higher than those of middle-aged patients in the Japanese Hypertension Society 2000 criteria, office and home BP values were not adequately controlled in approximately 50% of the middle-aged patients whose BP control was evaluated as good. Our findings suggest that an important reason why home and office BP values are not adequately controlled is that physicians approve relatively higher BP levels under treatment, even among middle-aged patients. PMID- 15785012 TI - Activation of extracellular signal-activated kinase by angiotensin II-induced Gq independent epidermal growth factor receptor transactivation. AB - Multiple signaling pathways link the angiotensin II (Ang II) type 1 (AT1) receptor to Gq-dependent inositol phosphate (IP) production and Gq-independent phospho-extracellular signal-activated kinase (p-ERK) 1/2 activation by Ang II in the regulation of cardiovascular vasoconstriction and cell growth, respectively. An Ang II analogue, [Sar1, Ile4, Ile8]Ang II, did not stimulate Gq-dependent IP production, but still activated Gq-independent p-ERK1/2 in human coronary artery smooth muscle cells as well as in a cell line that stably expressed AT1. This activation was mostly mediated by [Sar1, Ile4, Ile8]Ang II-induced Gq-independent epidermal growth factor receptor transactivation. We found that AT1 receptor signaling shows bifurcation into functionally separate pathways. A clear understanding of this unique signaling may be necessary for the development of therapeutic agents to treat disorders such as hypertension and cardiac hypertrophy. PMID- 15785014 TI - DNA extraction from bovine mummified fetuses and detection of factor XI gene deficiency in the mummies. AB - Genomic DNA extracted from bovine mummified tissue is valuable material for detection of some genes that may contribute to fetal abnormalities. In this study bovine genomic DNA was extracted from the hardened tissue samples of ten bovine mummified fetuses. The amount of genomic DNA extracted from 2 g of the mummified tissues by the phenol/chloroform-ethanol method was low (less than 4 microg/ml) for all samples. The extracted DNA was then amplified by the GenomiPhi DNA amplification system. After amplification, the amount of DNA was increased to more than 100 microg/ml for all samples. This amplification system was shown to be a good tool for amplifying the genomic DNA of the mummified fetuses. The amplified genomic DNA was used for testing the mummies for Factor XI gene deficiency, an autosomal recessive deficiency involved in the early stages of the intrinsic blood coagulation pathway. Exon 12 of the Factor XI gene of the mummies was amplified by PCR. Two of the ten mummified fetuses were heterozygous for the Factor XI gene as indicated by the presence of two amplified DNA fragments of 320 bp and 244 bp. Factor XI deficiency has already been described in Holstein cattle. However, no report is available for bovine fetus. In this study, DNA was extracted and amplified from the bovine mummified fetuses, and the samples were successfully tested for Factor XI gene deficiency in the mummies. PMID- 15785013 TI - Additive beneficial effects of the combination of a calcium channel blocker and an angiotensin blocker on a hypertensive rat-heart failure model. AB - The present study was undertaken to examine the effects of a calcium channel blocker, azelnidipine (1 mg/kg/day), an angiotensin converting enzyme (ACE) inhibitor, temocapril (10 mg/kg/day), an angiotensin II type 1 (AT1) receptor blocker (ARB), olmesartan (5 mg/kg/day), and their combination on Dahl salt sensitive rats (DS rats) developing heart failure with preserved systolic function. DS rats were fed a high-salt diet (8% NaCl) from 7 weeks of age and progressively developed hypertension. Although monotherapy with azelnidipine lowered the blood pressure of DS rats to a greater extent than monotherapy with temocapril or olmesartan, the three drugs had similar effects on cardiac hypertrophy, cardiac fibrosis, the expressions of brain natriuretic peptide, transforming growth factor-beta1, collagen I, collagen III and monocyte chemoattractant protein-1 mRNA (as estimated by Northern blot analysis), and cardiac diastolic dysfunction (as estimated by echocardiography). These results show that ACE and AT1 receptor, as well as hypertension, are involved in the development of heart failure with preserved systolic function in DS rats. The combination of azelnidipine with olmesartan or temocapril produced no additive hypotensive effect in DS rats and no additive effect on cardiac hypertrophy or gene expressions. However, the combination therapy prolonged the survival rate of DS rats more than azelnidipine (p <0.01) or temocapril alone (p <0.05), and this additive beneficial effect by the combination therapy was associated with a greater reduction of cardiac fibrosis, urinary albumin excretion and serum creatinine. Our results thus showed that the combination of a calcium channel blocker with an ARB or an ACE inhibitor had additive preventive effects on a rat model of hypertensive heart failure with preserved systolic function. Thus, combination therapy with these agents seems to be a useful therapeutic strategy for the prevention of hypertensive heart failure. PMID- 15785015 TI - Renal disease in cardiovascular disorders: an underrecognized problem. AB - Chronic renal disease is generally appreciated as a major and rapidly growing health problem. In the United States alone, as many as 19.5 million people may have markers of early renal disease, and more than 660,000 people are expected to require renal replacement therapy by the year 2010. By contrast, the presence and pathological role of renal disease in patients with cardiovascular disease are somewhat underrecognized. Evidence now shows that even minor impairments in renal function, as indicated by measures including glomerular filtration rate and microalbuminuria, are common in cardiovascular disease states and predictive of cardiovascular events. Indeed, microalbuminuria may be a marker of systemic vascular disease rather than kidney dysfunction alone. In patients with hypertension, diabetes, metabolic syndrome, acute coronary syndromes, and stroke, markers of renal disease have proved to be at least as predictive of morbidity and mortality as conventional risk factors. Yet, chart reviews in a variety of clinical settings reflect poor recognition and management of renal disease in at risk patients. Models for renal protection are based on the control of risk factors, particularly blood pressure, that are associated with renal and cardiovascular outcomes. Screening protocols for markers of renal disease should recognize the potential inaccuracy of serum creatinine concentrations and the preferability of glomerular filtration rate estimates that take age and gender into account. Pilot programs for screening high-risk populations have shown efficacy in detecting renal disease. PMID- 15785016 TI - Age- and sex-related variation in song-control regions in Cassin's finches, Carpodacus cassinii. AB - Male Cassin's finches (Carpodacus cassinii) sing long, complex songs that incorporate many elements mimicked from other species. Although one-year-old males (males in their first breeding season) are sexually competent and do breed, they sing a simpler song (fewer syllable types) than do males two or more years old (called after-second-year males). Females do occasionally sing, but with much less stereotypy and complexity than breeding males of any age. We collected brains from free-living breeding after-second-year males (identified by their red plumage), one-year-old males (identified by their female-like brown plumage), and adult females to examine sex- and age-related differences in three song-control nuclei: HVC, Area X and RA. Nuclei volumes were reconstructed by measuring the Nissl-defined area in every second section. There was a large sex difference in all three nuclei, with female volumes 40-50% that of males. There was no difference in HVC volume between age classes of males. However, one-year-old (brown) males had significantly larger Area X volume than did the older (red) males, with red males' Area X about 75% the volume of brown males'. These data raise questions regarding the functional significance of Area X and related nuclei in relation to song development in vocal mimics. PMID- 15785017 TI - Effects of intrauterine growth retardation on development of the gastrointestinal tract in neonatal pigs. AB - BACKGROUND: Intrauterine growth retardation (IUGR) is a common problem in human and other species and increases the risk of death of the fetus and newborn during the perinatal period. OBJECTIVES: This study was conducted to examine the influences of intrauterine growth retardation (IUGR) on development of the gastrointestinal tract in newborn pigs. METHODS: Ten animals from five litters were divided into five piglets with IUGR and five with normal birth-weight (NW). The IUGR category comprised animals with a birth weight 2 SD below the mean birth weight of the total population, while the NW category included animals with a birth weight within one SD of the mean birth weight in the total population. Animals were anesthetized and sampled within 2-4 h after birth and without suckling. The morphological changes of intestine and stomach of IUGR piglets were compared with NW ones. The expressions of IGF-I and receptors for growth hormone and insulin in intestinal mucosa were semiquantified using reverse transcription PCR. RESULTS: The results of our study indicated that the weights of the stomach, small intestine and small intestinal mucosa were significantly lower in IUGR compared with NW piglets (p<0.01). In addition, the lengths of the small intestine and colon in IUGR pigs were also significantly less than those of NW (p<0.05). Furthermore, insulin-like growth factor-I (IGF-I) mRNA level in intestinal mucosa of IUGR piglets was increased significantly (p<0.05), and the expression mRNA levels of insulin receptor and growth hormone (GH) receptor in the mucosa in IUGR piglets showed a tendency to be lower (p=0.17 and p=0.11, respectively) than those of the NW animals. CONCLUSION: We conclude from the data that IUGR affects intestinal growth and morphology and is in associated with altered gene expression of growth-related proteins. We speculate that the morphological change and associated altered endocrine homeostasis contribute to lower growth rates of pigs affected by IUGR. PMID- 15785018 TI - Effects of antenatal glucocorticoids on circulatory adaptation at birth in the ovine fetus. AB - OBJECTIVE: Adaptation to extra-uterine life requires dramatic increase in pulmonary blood flow. Mechanisms that induce pulmonary vasodilatation at birth are incompletely understood but include alveolar ventilation, increase in PaO2, and production of vasoactive mediators. We hypothesized that antenatal glucocorticoids (GC) increase pulmonary vasodilatation to birth-related stimuli. STUDY DESIGN: To test this hypothesis, we studied the pulmonary hemodynamic response at birth to mechanical ventilation with low (<10%) and then with high (100%) FiO2 in chronically prepared late-gestation fetal lambs treated or not by antenatal maternal steroids. RESULTS: Basal mean aortic and pulmonary artery pressure (PAP), left pulmonary blood flow, pulmonary vascular resistance (PVR), and blood gas were similar between control and dexamethasone-treated animals (GC group). During mechanical ventilation with low FiO2, mean PVR decreased by 40% in the control group (from 0.44 +/- 0.01 to 0.25 +/- 0.01 mm Hg/ml/min) and by 60% in the GC group (from 0.44 +/- 0.02 to 0.19 +/- 0.02 mm Hg/ml/min) (p < 0.01). When subsequently ventilated with 100% O2, there was no difference in PVR decrease between groups (0.15 +/- 0.02 mm Hg/ml/min in the GC group vs. 0.14 +/- 0.01 mm Hg/ml/min in the control group). CONCLUSION: Antenatal GC enhance pulmonary vasodilatation induced by alveolar ventilation at birth but do not alter the pulmonary vascular response to O2. We speculate that antenatal steroids exposure improve adaptation at birth through acceleration of both parenchymal and vascular lung maturation. PMID- 15785020 TI - Association of right ventricular dilatation with bilateral pulmonary embolism, pulmonary embolism in a main pulmonary artery and lobar, segmental and subsegmental pulmonary embolism in 190 patients with acute pulmonary embolism. AB - BACKGROUND: Acute pulmonary embolism (PE) may result in right ventricular (RV) pressure overload with a dilated RV which can be diagnosed by two-dimensional echocardiography. METHODS: A retrospective analysis was performed in 190 unselected patients who had acute PE documented by contrast-enhanced spiral computed tomographic scanning. The 190 patients included 104 women and 86 men, mean age 58 +/- 15 years. RESULTS: RV dilatation was present in 45 of 70 patients (64%) with bilateral PE, in 19 of 120 patients (16%) without bilateral PE, in 42 of 47 patients (89%) with main pulmonary artery embolism, in 34 of 84 patients (40%) with lobar PE, in 16 of 70 patients (23%) with segmental PE and in 6 of 36 patients (17%) with subsegmental PE; p < 0.001 comparing bilateral with no bilateral PE and main pulmonary artery embolism with no main pulmonary artery embolism, with lobar, segmental and subsegmental PE; p < 0.025 comparing lobar with segmental PE, and p < 0.02 comparing lobar with subsegmental PE. CONCLUSION: The prevalence of RV dilatation is highest in patients with main pulmonary artery embolism or bilateral pulmonary artery embolism; furthermore, the prevalence of RV dilatation is higher in patients with lobar PE than in patients with segmental or subsegmental PE. PMID- 15785019 TI - A comparison of the two beta-blockers carvedilol and atenolol on left ventricular ejection fraction and clinical endpoints after myocardial infarction. a single centre, randomized study of 232 patients. AB - BACKGROUND: beta-Blockers have been found to reduce mortality and morbidity in postmyocardial infarction patients. However, it is not fully understood whether all beta-blockers have similar favourable cardiovascular effects. The aim of this study was to compare the effects of carvedilol and atenolol on global and regional left ventricular ejection fraction (LVEF) and on predefined cardiovascular endpoints. METHODS: In a single-centre, randomized, open, endpoint blinded, parallel group study, 232 patients with acute myocardial infarction were randomized to treatment with carvedilol or atenolol. LVEF was measured by gated blood pool scintigraphy during the first week and after 12 months. The treatment was given orally within 24 h. The mean dose was 36.2 and 72.1 mg in the carvedilol and atenolol groups, respectively. RESULTS: No significant difference was found between the two study groups in the mean global and regional LVEF. There tended to be fewer first serious cardiovascular events in the carvedilol compared with the atenolol group (RR = 0.83, 95% CI 0.56-1.23, p = 0.39). Cold hands and feet were observed less frequently in the carvedilol group (20 vs. 33%, p = 0.025). CONCLUSION: In patients following an acute myocardial infarction, no difference in either global or regional LVEF was observed between baseline and 12 months when treatment with carvedilol was compared with atenolol. PMID- 15785021 TI - Association of ankle-brachial index with severity of angiographic coronary artery disease in patients with peripheral arterial disease and coronary artery disease. AB - The ankle-brachial index (ABI) was correlated with the severity of coronary artery disease (CAD) in 273 patients, mean age 71 years, with peripheral arterial disease and angiographically obstructive CAD (> 50% occlusion). Of 155 patients with an ABI < 0.40, 130 (84%) had 3- or 4-vessel CAD, 17 (11%) had 2-vessel CAD and 8 (5%) had 1-vessel CAD. Of 80 patients with an ABI of 0.40-0.69, 37 (46%) had 3- or 4-vessel CAD, 33 (41%) had 2-vessel CAD and 10 (13%) had 1-vessel CAD. Of 38 patients with an ABI of 0.70-0.89, 10 (26%) had 3- or 4-vessel CAD, 16 (42%) had 2-vessel CAD and 12 (32%) had 1-vessel CAD. The lower the ABI, the higher the prevalence of 3- or 4-vessel CAD and the lower the prevalence of 1 vessel CAD. PMID- 15785022 TI - Heart failure in diabetes mellitus: clinical features and prognostic implications. AB - We defined the prevalence and impact on survival of clinical bedside variables in 385 patients with symptomatic congestive heart failure (CHF), of whom there were 176 with and 209 without diabetes mellitus. Patients were consecutively hospitalized and admitted for various acute conditions. Following discharge all cause mortality was recorded. Prevalence and association of various variables with mortality were statistically analyzed. Prevailing in the diabetics versus nondiabetics were younger age (p < 0.05), pulmonary edema on admission (p = 0.002), using furosemide > 80 mg/day (p < 0.01) for > 1 year (p < 0.01) and hyponatremia (p = 0.01). Less prevalent were chronic lung disease (p < 0.01) and cardiac arrhythmias (p = 0.001). On follow-up extending up to 60 months, diabetic patients, especially those with fasting blood glucose levels on admission > or = 180 mg/dl, survived for a shorter period of time than nondiabetics (p = 0.02). Associated with increased mortality in the diabetic group were female gender (p = 0.04), furosemide > or = 80 mg/day (p < 0.001) and renal dysfunction (RD; p = 0.04). The respective variables in the nondiabetics were advanced age (p < 0.001) and RD (p = 0.002). Although they were younger, diabetic patients presented more severe CHF. It is recommended that special attention should be given to diabetic females, those using higher furosemide dosages and those suffering from RD. PMID- 15785023 TI - Noncompaction in myotonic dystrophy type 1 on cardiac MRI. PMID- 15785024 TI - Body temperature - a marker of infarct size in the era of early reperfusion. AB - We measured body temperature in 40 consecutive patients treated for a first ST elevation acute myocardial infarction (AMI) with primary percutaneous coronary interventions. Left ventricular function was assessed by echocardiography, and blood samples were drawn for highly sensitive C-reactive protein (hs-CRP), white blood cell (WBC) count, fibrinogen, creatine kinase (CK), and cardiac troponin I levels (cTnI). The median (25th, 75th quartiles) peak 24-hour temperature was 37.4 degrees C (36.9 degrees C, 37.6 degrees C). Variables significantly associated with peak 24-hour temperature were CK (p = 0.01, r = 0.42), wall motion index (p = 0.01, r = 0.41), hs-CRP (p = 0.01, r = 0.41), and cTnI (p = 0.03, r = 0.35). There was no significant correlation between peak 24-hour temperature and WBC count (p = 0.39, r = 0.14) and fibrinogen (p = 0.12, r = 0.21). Thus, peak 24-hour body temperature after ST elevation AMI probably reflects infarct size rather than a nonspecific inflammatory response. PMID- 15785025 TI - Study of cardiac rate and rhythm patterns in ambulatory and hospitalized children. AB - Our aim was to underline possible differences in heart rate and rhythm patterns between ambulatory and hospitalized children. Holter monitoring was performed on 264 healthy ambulatory children and on 112 children who were hospitalized for noncardiotoxic conditions. Maximal, mean and minimal heart rates decreased with age. Maximal heart rate was significantly higher in ambulatory schoolchildren and adolescents than in hospitalized ones. Sinus arrhythmia was noted on every recording. Some children had episodes of first- or second-degree atrioventricular block while sleeping. Supraventricular and uniform ventricular extrasystoles were common. The incidence of all types of arrhythmia and conduction disturbances was similar in ambulatory and hospitalized children. These data can be taken as a basis for the analysis of 24-hour electrocardiogram monitoring in ambulatory but also in hospitalized children. PMID- 15785026 TI - Levels of, and factors associated with, C-reactive protein in employees attending a company-sponsored cardiac screening program. AB - BACKGROUND: Relatively limited contemporary information is available about the distribution of, and factors associated with, levels of C-reactive protein (CRP) in adult men and women. The purpose of our descriptive study was to examine the prevalence and predictors of this marker of inflammation in a sample of employed adults attending a worksite cardiovascular screening program. METHODS: The study sample consisted of 876 men and women between the ages of 21 and 77 years from 6 locations of the parent company. These individuals attended an employer-sponsored cardiovascular screening and wellness program during 2003. A standardized questionnaire was administered to all study participants, and a number of different coronary risk factors were measured. RESULTS: Approximately 25% of the study sample was classified as having elevated CRP levels (> or =3 mg/l). Women, obese individuals, subjects with increasing heart rate and higher levels of serum triglycerides were more likely to have elevated concentrations of CRP than the corresponding comparison groups. Subjects who reported regularly exercising, individuals with a history of heart disease and those with lower total cholesterol levels were less likely to have elevated CRP levels. A relatively similar risk factor profile was noted in individuals without a self-reported history of prior cardiovascular disease. CONCLUSIONS: The results of our cross sectional observational study suggest that the prevalence of elevated CRP levels in the general adult population is considerable. A number of demographic, comorbid and other factors are associated with this inflammatory marker of increased risk of cardiovascular disease, which demands increased attention and modification of potential predisposing factors. PMID- 15785027 TI - Lack of association between common polymorphisms in genes of the renin angiotensin system and mortality after myocardial infarction. AB - The insertion/deletion (I/D) polymorphism in the ACE gene and the A1166C polymorphism in the AT1R gene have been associated with left ventricular remodelling and prognosis after acute myocardial infarction (AMI). We investigated whether these genetic variants associate with impaired left ventricular ejection fraction (LVEF) and increased risk for in-hospital mortality after AMI. Consecutive AMI patients were recruited on admission and were genotyped for the above-mentioned polymorphisms. The frequency of the studied genotypes did not differ significantly between deceased patients and those who survived. The LVEF did not differ among patients with or without the DD genotype (45 +/- 10 vs. 45 +/- 10%, p = 0.892) or the CC genotype (45 +/- 10 vs. 46 +/- 10%, p = 0.859). These data question the role of the studied genotypes in the pathogenesis of AMI and do not support the previously supported hypothesis that these genotypes influence prognosis after AMI. PMID- 15785028 TI - The effect of simvastatin treatment on the amyloid precursor protein and brain cholesterol metabolism in patients with Alzheimer's disease. AB - During the last years, several clinical studies have been published trying to elucidate the effect of statin treatment on amyloid precursor protein (APP) processing and metabolism of brain cholesterol in Alzheimer's disease (AD) in humans. We present an open biochemical study where 19 patients with AD have been treated with simvastatin (20 mg/day) for 12 months. The aim was to further investigate the effect of simvastatin treatment on cerebrospinal fluid (CSF) biomarkers of APP processing, AD biomarkers as total tau and tau phosphorylated at threonine 181, brain cholesterol metabolism as well as on cognitive decline in patients with AD. Despite biochemical data suggesting that treatment with 20 mg/day of simvastatin for 12 months does affect the brain cholesterol metabolism, we did not find any change in CSF or plasma levels of beta-amyloid (Abeta)(1-42). However, by analysis of APP isoforms, we found that statin treatment may favor the nonamyloidogenic pathway of APP processing. The relevance and mechanism between statin treatment and AD has to be further elucidated by using statins of different lipophility in different dosages over a longer period of time. PMID- 15785029 TI - Loss of cholinergic pathways in vascular dementia of the Binswanger type. AB - We sought to determine the changes in the cholinergic pathways, which project from the nucleus basalis of Meynert (nBM) and travel in the subinsular region, in vascular dementia of the Binswanger type (VDBT) and Alzheimer's disease (AD). The subinsular regions were examined in 6 autopsied brains with VDBT, 5 brains with AD and 4 control brains without any neurologic diseases. The cholinergic pathway was labeled either by histochemistry for acetylcholine esterase (AChE), a degradatory enzyme of ACh, or by immunohistochemistry for choline acetyltransferase, its synthetic enzyme. The numerical density of nBM neurons did not differ significantly between these groups (163 +/- 49 in the VDBT, 105 +/- 82 in the AD and 198 +/- 76 in the control groups), but with a tendency towards a decrease in the AD group. The subinsular cholinergic fibers were impaired, with relative preservation of the nBM neurons in VDBT, whereas both the subinsular cholinergic fibers and the nBM neurons were degraded in AD. These results indicate that the cholinergic pathway is damaged not only in AD, but also in VDBT, and may further provide a pharmacological basis for treatment with AChE inhibitors in VDBT. PMID- 15785030 TI - Motor function in subjects with mild cognitive impairment and early Alzheimer's disease. AB - Basic mobility, balance, gait and dual-task performance were characterised in 140 consecutive subjects referred to a multidisciplinary university hospital in a geriatric setting for cognitive symptoms and possible dementia. After completion of an extensive diagnostic evaluation, subjects were classified into four diagnostic categories: no cognitive impairment, mild cognitive impairment, Alzheimer's disease (AD) and other dementia. Mean age was 57 +/- 9.2, 60 +/- 7.3, 68 +/- 9.9 and 64 +/- 10.5, respectively. Data on motor function, medication use and presence of white matter changes were evaluated and compared between the diagnostic groups. Motor function seems to be affected in very mild AD but not in mild cognitive impairment, as assessed with performance-based tests. AD subjects were slowed and had difficulties in dual-task performance requiring concurrently performing a cognitive task while walking. PMID- 15785031 TI - Prevalence, resource utilization and costs of vascular dementia compared to Alzheimer's dementia in a population setting. AB - OBJECTIVES: To determine the prevalence of dementia and to measure the monetary impact and health resources utilization of vascular dementia (VD) compared to Alzheimer's dementia (AD) in persons aged over 64 years in a population setting. METHODS: Retrospective, cross-sectional study. In the initial phase, information was obtained on specific clinical characteristics from the subjects with an active diagnosis of dementia. The second phase consisted of a clinical evaluation and validation of the cases. Mini-Mental State Examination was used to assess cognitive impairment. Dementia and its subtypes were determined using established diagnostic criteria. Information was obtained on the use of health care resources (direct costs) and the number of hours devoted by the primary caregiver (indirect costs) for patients with a documented diagnosis of AD or VD within the last 6 months prior to the interview. A multiple logistic regression analysis was performed to correct the model. RESULTS: A total of 6,004 subjects were analyzed, 258 with diagnosis of dementia (overall prevalence: 4.3%). An evaluation was made of 224 patients, and gross prevalence of AD and VD was 2.4 and 1.0%, respectively. Cost per patient per semester was EUR 8,086 for AD and EUR 11,039 for VD (p = 0.016). 85.5% of the cost was attributable to primary caregiver time in AD and 84.4% in VD. CONCLUSIONS: The prevalence of AD and VD increases with age. No sociodemographic differences were seen between AD and VD. Costs associated with health care resource and primary caregiver utilization were high, being higher in VD than in AD. PMID- 15785032 TI - The impact of change in cognitive functioning and cognitive decline on disability, well-being, and the use of healthcare services in older persons. Results of Longitudinal Aging Study Amsterdam. AB - The study investigated the impact of change in cognitive functioning and cognitive decline on disability, well-being, and the use of healthcare services among older persons in the Longitudinal Aging Study Amsterdam (LASA). Data were collected from 1,349 subjects, aged 65-85 years, who had scores of 24 and higher on the Mini-Mental State Examination (MMSE) at baseline, over a period of 6 years in three waves. The results indicate that cognitive decline and changes in cognitive functioning in older persons who were either not impaired or only mildly cognitively impaired at baseline have an impact on disability, well-being, and the use of healthcare services. With the aging of the population, the number of persons with cognitive impairment is likely to increase, and appropriate services should be available to them. PMID- 15785033 TI - Diagnostic accuracy of mini-mental status examination and revised hasegawa dementia scale for Alzheimer's disease. AB - To compare the diagnostic accuracies of the Revised Hasegawa Dementia Scale (HDS R) and Mini-Mental Status Examination (MMSE) for Alzheimer's diseases (AD), we administered them simultaneously to 82 AD patients and 82 age- and sex-matched nondemented control subjects. The area under the receiver operator curve (AUC) for AD of the HDS-R (AUC(HDS-R)) and MMSE (AUC(MMSE)) were bigger than 0.90 indicating that both tests are useful for detecting AD. However, AUC(HDS-R) (0.952) was significantly larger than that of the AUC(MMSE )(0.902) regardless of the educational level of the subjects, indicating that the HDS-R is more accurate than MMSE in diagnosing AD. Moreover, the superiority of the HDS-R (AUC(HDS-R) = 0.894) to the MMSE (AUC(MMSE) = 0.704) remained significant in mild AD patients alone, who are the focus of screening. In conclusion, the HDS-R is better than the MMSE as a screening instrument for AD. PMID- 15785034 TI - Everyday memory impairment of patients with mild cognitive impairment. AB - We evaluated everyday memory impairment in 24 patients with mild cognitive impairment (MCI) with the Rivermead Behavioral Memory Test (RBMT) and compared the scores with those of 48 age-, sex- and education-matched normal controls (NC) and 48 age-, sex- and education-matched Alzheimer disease (AD) patients. Overall everyday memory was impaired in MCI patients but the severity was milder than that in AD patients. The MCI patients showed impairment of everyday memory tasks requiring delayed recall. But they could normally perform tasks immediately after memorizing, except for recalling and retracing a simple new route. The total Profile score correctly classified 100% of the MCI patients and 91.7% of NC, thus demonstrating the usefulness of the RBMT for diagnosing MCI patients. Prospective memory tasks were not useful for detecting the patients with MCI. PMID- 15785035 TI - Does Alzheimer's disease affect hippocampal asymmetry? Evidence from a cross sectional and longitudinal volumetric MRI study. AB - OBJECTIVE: To determine whether Alzheimer's disease (AD) is associated with preferential atrophy of either the left or right hippocampus. METHODS: We examined right-left asymmetry in hippocampal volume and atrophy rates in 32 subjects with probable AD and 50 age-matched controls. Hippocampi were measured on two serial volumetric MRI scans using a technique that minimizes laterality bias. RESULTS: We found a non-significant trend for right > left (R > L) asymmetry in controls at both time points (R > L: 1.7%; CI: -0.3-3.7%; p = 0.1). AD subjects showed a similar non-significant trend for R > L asymmetry at baseline (R > L: 1.8%; CI: -1.9-5.5%; p = 0.32), but not at repeat (p = 0.739). Change in R/L ratio between visits in AD patients was significant (p = 0.02). The AD group had significantly higher variance in these ratios than the controls at baseline (p = 0.02), but not repeat (p = 0.06). AD patients had higher atrophy rates than controls (p < 0.001). Mean (CI) annualized atrophy rates for left and right hippocampi were 1.2% (0.5-1.8%) and 1.1% (0.5-1.8%) for the controls, and 4.6% (3.3-6.0%) and 6.3% (4.9-7.8%) for AD subjects. There was no significant asymmetry in atrophy rates in controls (p = 0.9), but borderline significantly higher atrophy rates in the right hippocampus of the AD group (p = 0.05) compared to the left. Presence of an APOEepsilon4 allele had no significant effect on the size, asymmetry or atrophy rates in AD (p > 0.20). CONCLUSIONS: We report minor R > L asymmetry in hippocampal volumes in controls and present some evidence to suggest that there is a change in the natural R > L asymmetry during the progression of AD. PMID- 15785036 TI - Cholinesterase inhibitor affects the amyloid precursor protein isoforms in patients with Alzheimer's disease. AB - An altered platelet ratio of amyloid precursor protein (APP) isoforms might be a diagnostic, predictive, or therapeutic marker for Alzheimer's disease (AD). Our purpose was to test the hypothesis that this ratio might serve as a therapeutic marker for AD patients treated with the cholinesterase inhibitor, galantamine. Thirty-nine patients (mean age 76.6 +/- 9.4 years) with AD were treated with galantamine for 12 weeks. Patients were evaluated at baseline, 4 and 12 weeks by cognitive testing along with a determination of their platelet APP isoform ratio. Western blotting was performed to calculate the APP isoform ratio. At the end of the treatment, cognitive scores significantly improved, and the ratio of the high molecular-weight (130 kDa) isoform to the low-molecular-weight (110-106 kDa) isoforms increased. These results suggest that cholinesterase inhibition might be involved in APP processing. PMID- 15785037 TI - Gastric perception thresholds are low and sensory neuropeptide levels high in helicobacter pylori-positive functional dyspepsia. AB - BACKGROUND AND AIMS: In functional dyspepsia (FD) decreased perception levels can be shown on gastric distension. Substance P (SP) and calcitonin gene-related peptide (CGRP) are involved in the sensitization of afferent neuronal pathways due to chronic inflammation. The role of Helicobacter pylori-induced gastric mucosal inflammation in the pathogenesis of FD is controversial. The aim of this study was to assess whether FD patients have altered mucosal concentrations of CGRP and SP, and to investigate whether this is associated with visceral hypersensitivity or H. pylori infection. METHODS: Gastrointestinal symptoms, H. pylori status, perception thresholds at gastric balloon distension, and gastric mucosal concentrations of CGRP and SP were determined in 13 FD patients and 18 healthy controls (HC). RESULTS: In H. pylori-positive FD patients discomfort and pain thresholds on gastric distension were lower compared to other groups. Antral mucosal levels of CGRP and SP were higher in H. pylori-positive subjects. In FD significantly negative correlations between discomfort and pain thresholds and antral mucosal concentrations of CGRP and SP were observed. CONCLUSIONS: In FD low perception thresholds on gastric distension are associated with high levels of CGRP and SP in the antrum, suggesting that sensory neuropeptides are involved in FD pathophysiology. PMID- 15785038 TI - Matrix metalloproteinases 2 and 9 are markers of inflammation but not of the degree of fibrosis in chronic hepatitis C. AB - BACKGROUND: The degree of liver fibrosis and inflammation is important in patients with chronic hepatitis C (CHC) in terms of therapy as well as prognosis. To obviate the need of liver biopsy, serum markers such as procollagen I, III and hyaluronic acid have been proposed but were found to be inaccurate. Controversy still exists regarding the role of matrix metalloproteinases (MMPs) as valid markers of liver fibrosis. AIM: To assess liver and serum MMP-2 and -9 as markers of fibrosis and inflammation in patients with CHC. METHODS: Thirty-five CHC patients and 8 non-hepatitis C patients with normal liver enzymes underwent liver biopsy. Activities of inflammation and fibrosis stage were determined by the Desmet score on a scale of 0-4. Serum and liver tissue MMP-2 and -9 activities were measured by zymography using substrate impregnated gels. RESULTS: Patient and control groups were similar in terms of age (50.8 +/- 15.1 vs. 50.6 +/- 15.2) and male/female ratio (18/17 vs. 4/4). In serum, MMP-9 activity was increased in patients compared to controls (308 +/- 110 vs. 163.5 +/- 35 , p < 0.05). In liver tissue, MMP-9 was also higher in patients than in controls (21 +/- 4.5 vs. 17.1 +/- 5.1, p < 0.05), whereas MMP-2 did not differ between patients and controls. Serum MMP-9 values correlated with liver histologic inflammatory grade (290.4 +/- 83 in grade 2 vs. 562.1 +/- 128 in grade 3, p < 0.05) but not with fibrosis stage. The highest rising in serum MMP-9 levels was observed between grade 2 to grade 3 and was superior to the rising in serum transaminase levels, indicating its advantage in assessing the progression of disease activity. No correlation between liver MMP activities and liver fibrosis or inflammation was observed. CONCLUSION: Serum MMPs, in particular MMP-9, can serve as markers of disease activity rather than fibrosis stage in chronic HCV patients. PMID- 15785039 TI - Effects of the tyrosine kinase inhibitor imatinib on neuroendocrine tumor cell growth. AB - AIM: We investigated the effects of the tyrosine kinase inhibitor imatinib (Gleevec) on neuroendocrine tumor cells. METHODS: Neuroendocrine tumor cells were incubated without and with imatinib. The effects on growth were examined by methylthiazoletetrazolium (MTT) assay. The c-Kit expression in human endocrine tumor tissue and cell lines was determined by immunohistochemistry and Western blot analysis, respectively. Cytotoxicity assay was performed by fluorescence activated cell sorting. The telomerase activity was determined using the telomeric repeat amplification protocol. RESULTS: 28% of the insulinomas, 100% of the gastrinomas, and 38% of the carcinoids expressed c-Kit. Imatinib at concentrations >5 microM inhibited cell proliferation and induced apoptosis in both c-Kit-positive and c-Kit-negative cell lines. The PI-3K inhibitor wortmannin did not enhance the effects of imatinib. Imatinib did not sensitize endocrine tumor cells to doxorubicin and 5-fluorouracil. Imatinib inhibited the telomerase activity. CONCLUSION: Imatinib inhibits neuroendocrine tumor cell growth independently of c-Kit by inhibition of other tyrosine kinases or through tyrosine kinase independent pathways. PMID- 15785040 TI - Management of severe refractory adult autoimmune enteropathy with infliximab and tacrolimus. AB - INTRODUCTION: Autoimmune enteropathy (AIE) is a rare cause of intractable diarrhea in infancy. A variant with onset in adulthood has recently been described. The immune mechanisms underlying AIE are unclear and the experience in the management of adult AIE is very limited. CASE REPORT: A 19-year-old male patient with massive refractory diarrhea and significant weight loss was diagnosed with AIE. He did not improve with corticosteroid and azathioprine treatment and was then treated successfully with infliximab. The response to treatment was rapid and complete. Because of a severe life-threatening hypersensitivity reaction during the 4th infliximab dose, infliximab had to be discontinued and therapy with tacrolimus was initiated. A complete clinical remission has since been maintained. CONCLUSION: Both infliximab and tacrolimus may provide a therapeutic alternative in patients with adult refractory AIE. PMID- 15785041 TI - Tacrolimus and the pigmentary system. PMID- 15785042 TI - A new era in the management of psoriasis? Promises and facts. AB - A number of new systemic agents, collectively termed as 'biologicals,' are made available for the treatment of psoriasis. We can display cautious optimism as to the role of these new agents, provided that further studies will be conducted in reasonable time. Rates of relapse on drug withdrawal, predictive factors for treatment failure and extensive data on drug safety are additional pieces of information required. Comparative effectiveness is also of concern. Advancements in clinical research, including severity assessment and outcome measures, should parallel improvements in the understanding of the pathomechanisms of psoriasis. A number of controversial areas exist concerning the management of psoriasis, where no direct interest from pharmaceutical companies is involved. The role of interventions like education and psychological support as an adjunct to conventional treatment is just one example. Independent research networks may be required to address these controversial areas. PMID- 15785043 TI - 'Atypical' cases of hailey-hailey disease may represent typical examples of a type 2 segmental manifestation. PMID- 15785044 TI - Silicon image sensor technology for in vivo detection of surfactant-induced corneocyte swelling and drying. AB - BACKGROUND: Several instrumental methods can indirectly assess some specific aspects of cutaneous irritation at the level of the stratum corneum (SC). OBJECTIVE: There is a need for developing more sensitive approaches in this field. METHODS: We assessed a recently introduced innovative tool (SkinChip) based on capacitive pixel-sensing technology in its potential to detect early discrete manifestations of skin irritation. The sensor generates a detailed non optical picture corresponding to a capacitance map of the skin surface reaching 50 microm pixel resolution. Some topographical details can be easily disclosed and the SC hydration as well. Two surfactant solutions were tested on volunteers. These solutions were applied under test patches for 2 days on the volar forearms. Clinical and SkinChip assessments were performed 3 h after removing the patch. RESULTS: The generated images allowed a precise observation of skin irritation which appeared as a two-step process. Early changes consisted of darker pixels corresponding to overhydrated swollen corneocytes at the irritated sites. Two days later, the same area appeared as white pixels, indicating the loss of corneocyte hydration. CONCLUSION: The SkinChip device appears to be a very sensitive tool for detecting the early steps of surfactant-induced skin irritation affecting the SC. PMID- 15785045 TI - Vascularity and fractal dimension of the dermo-epidermal interface in guttate and plaque-type psoriasis. AB - BACKGROUND: Histological structures of the skin are often irregular in size and shape. Euclidean geometry and fractal analysis are complementary for assessing distinct aspects of their dimensions. OBJECTIVE: To determine and compare the variations in shape of the dermo-epidermal junction and the size of the superficial vessels in psoriatic lesions. METHOD: The relative microvasculature area and the fractal dimension D of the dermo-epidermal interface were measured inside and outside growth-stunted guttate lesions (n = 22) and expanding plaques (n = 37) in psoriasis of the trunk. RESULTS: The median D values of the dermo epidermal interface were significantly larger (p < 0.01) in psoriatic plaques (D = 1.15) than in guttate lesions (D = 1.08), and these D values on lesional skin were significantly larger (p < 0.01) than in the uninvolved skin (D = 1.03). The microvasculature was significantly (p < 0.01) more developed in lesional (plaque: 13%, guttate: 8.20%) than in uninvolved skin (3.60 and 3.85%). No correlations were found between the relative microvasculature areas and the D values of the dermo-epidermal interface, both in the uninvolved and lesional skins of each psoriatic type. CONCLUSION: The absence of a relationship between modulations of the dermo-epidermal junction and vascular hyperplasia, both in expanding and stable psoriasis lesions, suggests that these events are regulated by different mechanisms and do not depend on each other. PMID- 15785046 TI - The psoriasis area and severity index is the adequate criterion to define severity in chronic plaque-type psoriasis. AB - BACKGROUND: Chronic plaque-type psoriasis is a major dermatosis, but a significant question is still unanswered: What defines severity in chronic plaque type psoriasis? While objective assessments like the Psoriasis Area and Severity Index (PASI) have frequently been used in clinical trials, quality of life (QOL) questionnaires are currently becoming more and more popular. OBJECTIVE: This article summarizes the most important objective and subjective measurements of severity in psoriasis. For every dermatologist it is critically important to distinguish between severe psoriasis and psoriasis that severely affects QOL. Even if the PASI also has disadvantages, it is the most adequate instrument available to evaluate severity in plaque-type psoriasis. RESULT: We provide reasons why PASI >12 defines severe, PASI 7-12 moderate and PASI <7 mild chronic plaque-type psoriasis. PMID- 15785047 TI - Targetoid hemosiderotic nevus. A trauma-induced simulator of malignant melanoma. AB - BACKGROUND: Simulators of malignant melanoma comprise a heterogenous group of melanocytic and nonmelanocytic lesions of the skin. Among frequent clinical mimickers of melanoma are injured melanocytic nevi. Any change in the clinical appearance of a pre-existing nevus should alert the clinician to exclude the possibility of malignant transformation in order to early identify a lesion at a stage when complete cure can still be achieved. OBJECTIVE: The purpose of this study was to present the clinical, dermoscopic and histopathologic findings of a series of acquired melanocytic nevi which abruptly developed a pigmented peripheral halo, presumably following minor trauma. METHODS: A series of 6 cases of acquired melanocytic nevi which suddenly developed a targetoid halo were included in the study. All lesions were evaluated by dermoscopy. Three cases were surgically removed at different stages of evolution and submitted to histopathologic examination. In all cases, follow-up was obtained. RESULTS: All the lesions arose on trauma-prone skin sites of young women. The sudden development of an asymptomatic, targetoid halo on a long-lasting, acquired exophytic nevus was the main presentation. Whereas the central nevus persisted, the ecchymotic halo ultimately disappeared. Histopathologic examination disclosed changes of the traumatized nevus in the central part, whereas the ring showed hemorrhage and hemosiderin deposits. Increased numbers of small vessels with hobnail characteristics were associated features. CONCLUSIONS: Targetoid hemosiderotic nevus is a distinctive clinicopathologic variant of traumatized acquired melanocytic nevus which should be included in the list of clinical simulators of melanoma. PMID- 15785048 TI - Changing trends in the number of deaths from nonmelanoma skin cancer in Japan, 1955-2000. AB - BACKGROUND: The incidence and number of deaths from nonmelanoma skin cancer (NMSC) have been reported to increase in many countries. However, those in Japan over a long time span have not been analyzed yet. OBJECTIVE: To analyze the changing trends in the numbers of deaths from NMSC in Japan. METHODS: We analyzed the annual trends in numbers of deaths from NMSC from 1955 to 2000 in Japan on the basis of the data from the Vital Statistics of Japan, Statistics and Information Department, Minister's Secretariat, Ministry of Health, Labor and Welfare. RESULTS: The general trends in numbers of deaths from NMSC were downward between 1979 and 1994, but upward after 1995. The recent annual increased proportion of deaths from NMSC was 3.8% (95% confidence interval: 1.3-6.3%). The general trends in age-standardized death rates were roughly downward, although the death rates plateaued after 1995. The number and proportion of deaths among the elderly were increasing. In recent years, more than 50% of the deaths among females occurred at or after 85 years of age, and the corresponding age for males was 75 years or after. CONCLUSION: Elderly individuals should be a focus of health promotion activities. PMID- 15785049 TI - Wound teleconsultation in patients with chronic leg ulcers. AB - BACKGROUND: The treatment of chronic leg ulcers requires frequent assessments of local wound status and adjustment of therapy. The availability of reasonably priced photographic equipment and quick electronic transfer of high-quality digital images should make it possible that the assessment of wound status can be made by remote experts. OBJECTIVE: This study examines the feasibility of using teledermatology for wound assessment and therapeutic suggestions for patients with chronic leg ulcers. METHODS: One hundred ten chronic leg ulcers of different origins were examined in face-to-face consultations. The examining doctor assessed the wound, made therapeutic recommendations and took 1-4 photographs of the wound using a digital camera. The digital images and relevant clinical information were then transmitted via a web application to an expert in wound care, who provided an independent teledermatological assessment of wound status and therapeutic recommendations. RESULTS: In our study, a high accordance between direct consultations and electronic consultations was found in the assessment of chronic leg ulcers, especially for important features like slough (concordance: 84.6%), necrosis (concordance: 98.2%) and granulation tissue formation (concordance: 76.4%). Furthermore, the teledermatologist generally felt confident in recommending further treatment strategies and in planning further wound assessments via the internet. CONCLUSIONS: Our results suggest that teledermatology offers great potential for the future in chronic wound care. By reducing the need to travel long distances to the hospital or to consult a physician with expertise in wound care, wound teleconsultation might lower health care costs and improve the quality of life for patients with chronic wounds, while still maintaining a high quality of wound care. PMID- 15785050 TI - The outcome after cryosurgery and intralesional steroid injection in vulvar lichen sclerosus corresponds to preoperative histopathological findings. AB - BACKGROUND: Vulvar lichen sclerosus (LS) still presents a therapeutic challenge. QUESTIONS: The aim of the present study was to investigate whether there is a correlation between preoperative histopathological findings in female LS and long term results after cryosurgery and intralesional steroid injections. MATERIAL AND METHODS: 22 females with LS, mean age 65.3 +/- 10.5 years, were treated with cryosurgery (-186 degrees C) using the contact method with 1 freeze-thaw cycle per lesion and intralesional triamcinolone acetonide injection. The time between surgery and follow-up examination was 27.8 +/- 14.6 months. We compared the clinical outcome and the thickness of the epidermis, the zone of sclerosis and the inflammatory infiltrate in biopsy specimens taken before treatment. RESULTS: In 14 of 22 patients, a relapse after cryosurgery occurred (median after 7.5 months). Patients would 'moderately' recommend cryosurgery for LS (2.2 +/- 0.9; minimum: 0, optimum: 4). Pruritus was significantly reduced (visual analog scale: preoperative 7.1 +/- 2.6 vs. postoperative 2.0 +/- 2.4; p = 0.001) even in patients with relapse (preoperative 7.6 +/- 2.4 vs. postoperative 3.1 +/- 2.28; p = 0.001). Women with relapse showed a significantly higher composed thickness of epidermis, sclerosis and inflammatory infiltrate (430 +/- 139.0 microm) than those without relapse (314 +/- 41.2 microm; p = 0.046). CONCLUSION: In our patients, cryosurgery and intralesional steroid injections were efficient in the treatment of LS. Patients showing relapse had a thicker epidermis and a stronger inflammatory infiltrate. Therefore a treatment of LS should be performed at an early stage of the disease. PMID- 15785051 TI - Disseminated granuloma annulare--treatment with fumaric acid esters. AB - BACKGROUND: The therapy of disseminated granuloma annulare has often limited success. Treatment of granuloma annulare with fumaric acid esters (FAE) has recently been reported to be effective in 2 patients. OBJECTIVES: To assess the efficacy of a systemic therapy with FAE in consecutive patients with disseminated granuloma annulare. METHODS: Eight patients with disseminated granuloma annulare were treated with FAE in tablet form according to the standard therapy regimen used in psoriasis. The colour and the elevation of the skin lesions were assessed by a visual analogue scale before and after therapy. RESULTS: Systemic therapy with FAE induced a significant clinical improvement in elevation and colour of skin lesions, with remission in 3 and partial remission in 4 patients. One patient remained unchanged. Side-effects associated with the therapy were seen in 6 patients. CONCLUSIONS: Systemic therapy with FAE can be effective in patients suffering from disseminated forms of granuloma annulare, but side-effects of FAE have to be taken into consideration. PMID- 15785052 TI - Alopecia areata universalis sparing nevus flammeus. AB - Alopecia areata is a common form of nonscarring hair disorder of unclear etiology. Cases with alopecia areata sparing congenital melanocytic nevi have been described. It is explained by the Renbok phenomenon or inverse Kobner phenomenon, which refers to normal hair growth in the psoriatic lesions observed in patients with psoriasis and alopecia areata. We report a 30-year-old man presenting with rapidly progressing alopecia areata universalis, which spared a large nevus flammeus on the left parietal scalp. Our case gives further evidence to the notion that nevus as genetic mosaic may show genetically determined resistance to alopecia areata. PMID- 15785053 TI - Mucosal pigmentation after oral lichen planus treatment with topical tacrolimus. AB - Erosive oral lichen planus (OLP) is a painful chronic inflammatory disease that is sometimes resistant to systemic or topical therapies. Topical steroids remain the mainstay of therapy, but topical tacrolimus has recently been used to treat OLP resistant to topical corticosteroids. Topical tacrolimus appears as an effective and safe treatment of symptomatic OLP. We report the first histopathologically documented case of oral mucosa pigmentation after OLP treatment with topical tacrolimus. The relation between tacrolimus treatment and staining was suggested by the appearance of pigmentation during topical tacrolimus treatment and its clinical disappearance when treatment was stopped. Histopathology showed an increase in melanocyte numbers and melanogenesis. PMID- 15785054 TI - Malignant melanoma in a candidate for heart transplantation. AB - A superficial spreading melanoma (Breslow thickness 0.4 mm) was diagnosed in a 65 year-old candidate for heart transplantation due to refractory end stage heart failure. After extensive review of the literature (USA and Europe), no clear guidelines about the management of candidates for transplantation with a previous diagnosis of melanoma were found. As this patient had a 5-year probability of survival higher than 95% and heart transplantation was necessary for saving his life, the final decision was to perform the transplantation. Unfortunately, the patient died of heart failure before a suitable heart became available. This case stresses the need for early and continuous dermatological evaluation of all candidates for solid organ transplantation. Clear guidelines for screening of skin cancer before transplantation are needed. PMID- 15785055 TI - Detection of doxorubicin in the horny layer in a patient suffering from palmar plantar erythrodysaesthesia. PMID- 15785056 TI - Eosinophilic pustular folliculitis with polycythemia vera. PMID- 15785057 TI - Early videocapillaroscopic changes of the psoriatic skin after anti-tumour necrosis factor alpha treatment. PMID- 15785058 TI - Calcium acetate-induced linear fixed drug eruption. PMID- 15785059 TI - Urticarial reaction to the antihistamine levocetirizine dihydrochloride. PMID- 15785060 TI - Interstitial pneumonitis and lung fibrosis during adjuvant treatment of melanoma with interferon alpha according to the Kirkwood schedule. PMID- 15785061 TI - Cutaneous leishmaniasis caused by Leishmania tropica: treatment with oral fluconazole. PMID- 15785062 TI - Subacute cutaneous lupus erythematosus associated with tamoxifen therapy: two cases. PMID- 15785063 TI - Methotrexate use and monitoring in Switzerland and Denmark. PMID- 15785064 TI - Sea urchin granuloma. PMID- 15785065 TI - Success of a smoking cessation program among smoking relatives of patients with serious smoking-related pulmonary disorders. AB - The aim of the study was to investigate the success of a smoking cessation program among the smoking relatives of patients with serious smoking-related disorders and to determine whether having a relative with such a disorder influences the rate of success of the program. The relatives of patients with chronic obstructive pulmonary disease (COPD) and lung cancer who wanted to quit smoking were included in the smoking cessation program (group I, n = 34). During the same period, the same number of subjects with relatives free of any smoking related diseases were included as a control group (group II, n = 34). Intensive behavioral therapy and motivational support was given to all subjects. Nicotine replacement therapy was administered to 10 subjects (29.4%) in group I and 14 subjects (41.2%) in group II. The subjects were evaluated in the 1st, 2nd, 4th, 8th and 12th week and subsequently every 3 months for a year in the outpatient clinic and proactive telephone interviews were performed periodically. In group I, 23.5% of subjects had a relative with COPD, 61.8% had relatives with lung cancer, and 14.7% had relatives with both COPD and lung cancer. The rate of quitting smoking was 79.4 and 73.5% on the 15th day and 41.2 and 38.2% at the end of the 1st year in groups I and II, respectively (p > 0.05). We observed that having a relative with serious smoking-related disorders did not motivate smokers to quit smoking. PMID- 15785066 TI - Reliability and validity of the French version of the tobacco craving questionnaire. AB - OBJECTIVES: (1) To assess the factorial structure, reliability and validity of the French language version of the Tobacco Craving Questionnaire (TCQ), a multidimensional, self-report instrument evaluating tobacco craving, to allow its use in French-speaking populations. (2) To compare the two different language versions of the same instrument in 2 independent samples from 2 cultures in 2 distinct time periods. METHODS: The French TCQ was administered to 226 current cigarette smokers after at least 2 h of abstinence. RESULTS: Despite the substantial difference in population characteristics between the French and American samples of smokers, both exploratory and confirmatory factor analyses indicated that the French version was best described by a 4-factor solution, similar to the original English version. Reliability estimates were 0.83, 0.79, 0.69 and 0.66 for the French and 0.82, 0.70, 0.75 and 0.48 for the English version for 'emotionality' (factor 1), 'expectancy' (factor 2), 'compulsivity' (factor 3) and 'purposefulness' (factor 4), respectively. The 4 factors accounted for 11.6, 17.1, 9.0, and 9.5% of the total variance in emotionality, expectancy, compulsivity, and purposefulness, respectively, for the French compared to 14.2, 12.5, 14.4 and 5% of the total variance for the English version, respectively. CONCLUSION: The French version of the TCQ is as valid and reliable instrument as the original English version and assesses the same 4 dimensions of craving for tobacco. PMID- 15785067 TI - Gender differences in health-related quality of life on admission to a maintenance treatment program. AB - We compared the gender differences in health-related quality of life (QOL) on admission to a maintenance program. 103 opioid users (65 men and 38 women) admitted to a maintenance treatment program during 2000-2002 were studied. During this period we assessed the QOL status using the German version ('Berlin Quality of Life Profile') of the Lancashire Quality of Life Profile. Physical symptoms were measured using the Opioid Withdrawal Scale. 312 urine screening tests were carried out to evaluate consumption. The female group showed significantly less additional consumption of other opiates (p = 0.043) compared with the male group. The male group showed significantly better QOL scores in self-esteem (p = 0.015), psychical health (p = 0.027), and law and security (p = 0.008). The outcome measures for withdrawal scores showed significantly less symptoms for males in twitching of muscles (p = 0.034), vomiting (p = 0.002), depressions (p = 0.004) and poor appetite (p = 0.008). In summary, both genders showed only a few significant differences on admission in terms of QOL and physical symptoms. The predominant effects of drug use appear to eclipse the gender-related role pattern. Further exploration of gender and QOL could have important theoretical and treatment implications. PMID- 15785068 TI - The unique role of diamorphine in British medical practice: a survey of general practitioners and hospital doctors. AB - AIMS: Diamorphine (heroin) is one of the longest established medications with a 130-year history. In British medicine diamorphine is sometimes used as a maintenance treatment for opiate addiction but is also routinely used in clinical practice to treat a number of general medical conditions. These uses are very different but are often confused by observers of the British system. The routine clinical use of diamorphine in medicine is unique to the UK, but this is not well known outside the UK. The present study investigates the use of diamorphine to treat medical conditions by general practitioners and by hospital doctors. METHOD: In a survey of 141 medical practitioners who had prescribed diamorphine for the treatment of a medical condition, data were collected on conditions prescribed for, numbers of patients, dosage, routes of administration, duration of treatment, worries about prescribing diamorphine and observed adverse effects. FINDINGS: Many doctors had prescribed diamorphine to a large number of patients. The four main conditions for which diamorphine had been prescribed in the previous year were myocardial infarction, palliative care, pulmonary oedema, and post-operative pain. More than half of the sample (57%) reported no reservations about prescribing diamorphine. Of the doctors (n = 60, 43%) who expressed worries, this was most often about possible problems of respiratory depression or respiratory arrest. Respiratory arrest/failure was also the most frequent observed adverse effect. Relatively few doctors reported worries about addiction. CONCLUSIONS: Although diamorphine was widely used and most doctors regarded it as a useful medication, there remains insufficient research information about current clinical practice. At a time when diamorphine may be coming under increased scrutiny, more detailed information is required of its uses and applications. PMID- 15785069 TI - Pharmacological relapse prevention of alcoholism: clinical predictors of outcome. AB - OBJECTIVE: The efficacy of pharmacological relapse prevention in alcoholism with acamprosate and naltrexone has been supported by several controlled trials. It remains uncertain whether any differential indication for treatment exists. METHODS: We evaluated outcome data of a controlled trial on acamprosate and naltrexone in patients with low vs. high baseline somatic distress, depression and anxiety (Symptom Checklist-90, SCL-90), low vs. high baseline craving, and according to typological differentiation as proposed by Cloninger and Lesch. These variables have previously been suggested to be predictors of outcome. RESULTS: Comparing the course of abstinence rates, acamprosate was mainly efficacious in patients with low baseline somatic distress, whereas naltrexone was effective especially in patients with high baseline depression. Baseline craving showed no predictive value. Pharmacological treatment was efficacious in type II alcoholics according to Cloninger. Applying Lesch's typological differentiation, acamprosate was shown to be mainly effective in type I, whereas naltrexone revealed best treatment effects in type III and IV. CONCLUSION: The study supports the hypothesis that different subgroups of alcohol dependent subjects might benefit from a differential treatment with either naltrexone or acamprosate. Baseline psychopathology and especially typological differentiation might be useful in matching patients to distinct pharmacotherapeutic interventions. . PMID- 15785070 TI - Association between early onset of cigarette, alcohol and cannabis use and later drug use patterns: an analysis of a survey in European metropolises. AB - OBJECTIVES: The study presents data about age of onset of alcohol, cigarette and cannabis use and investigates the association between age of onset and later drug use patterns. METHODS: Using a sample from a cross-sectional multi-site study, personal interviews were conducted with 3,503 individuals aged 12-49 years. Last month prevalence, age of onset and associations with subsequent use patterns were investigated. RESULTS: Having started with cannabis before the age of 16 years was associated with an odds ratio of 1.6 for heavy cannabis use. For males, the odds ratio of heavy cannabis use was 1.7, when cannabis was already initiated by the age of 16. Heavy use of ecstasy, amphetamines, hallucinogens and cocaine is associated with use of cigarettes before 13 (OR = 1.9). For males, the odds ratio was 2.2 and for women 1.9. CONCLUSIONS: Early use of alcohol, according to this data, does not seem to be related to subsequent heavy drug use. Early onset cannabis users show increasing probabilities of heavy use patterns. Preventive intervention programs have to start earlier than school-based programs normally do and specific developmental pathways need to be addressed. PMID- 15785071 TI - Methadone maintenance treatment in the Swiss Canton of Vaud: demographic and clinical data on 1,782 ambulatory patients. AB - Using data from the Public Health Service, we studied the demographic and clinical characteristics of 1,782 patients enrolled in methadone maintenance treatment (MMT) during 2001 in the Swiss Canton of Vaud, comparing our findings with the results of a previous study from 1976 to 1986. In 2001, most patients (76.9%) were treated in general practice. Mortality is low in this MMT population (1%/year). While patient age and sex profiles were similar to those found in the earlier study, we did observe a substantial increase in the number of patients and the number of practitioners treating MMT patients, probably reflecting the low-threshold governmental policies and the creation of specialized centers. In conclusion, easier access to MMT enhances the number of patients, but new concerns about the quality of management emerge: benzodiazepine as a concomitant prescription; low rates of screening for hepatitis B, C and HIV, and social and psychiatric preoccupations. PMID- 15785072 TI - MR-pathologic comparison of the upper spinal cord in different motor neuron diseases. AB - This MRI study was performed to evaluate in vivo alterations of the spinal cord in defined subgroups of motor neuron diseases. Standard MRI examinations of the cervical and thoracic spinal cord in sporadic amyotrophic lateral sclerosis (ALS; n = 39), sporadic lower motor neuron disease (LMND; n = 19), Kennedy's disease (KD; n = 19) and a control group (n = 96) were analyzed with respect to spinal cord signal changes and the thickness of the spinal cord. No significant changes in thickness or signal alterations were observed when comparing ALS, LMND and control groups with one another. However, in KD patients significant upper spinal cord atrophy was detected at the cervical level as compared with all other groups. At the thoracic level, KD patients had significant upper cord atrophy as compared with controls and LMND. Marked atrophy of the upper spinal cord seems to be a feature of the KD-associated central-peripheral distal axonopathy. PMID- 15785073 TI - Cough headache and thoracic inlet valvular competence in uremia. AB - Cough headache is a transient headache upon coughing, bending, stooping, or lifting in the absence of intracranial lesions. Reports show that incompetent jugular venous valve and cerebrospinal fluid hypervolemia are contributing factors. Headache is a common complaint of uremia patients. We conducted a clinical-radiological correlation study on 15 uremia patients with headache and central venous thrombosis. Thirteen patients were diagnosed to have benign cough headache (BCH); the others were diagnosed with chronic tension type headache. Venogram disclosed either internal jugular or vertebral venous regurgitation in the BCH group. Acquired thoracic inlet valvular incompetence might contribute to BCH. PMID- 15785074 TI - Luteal phase support in in vitro fertilization: meta-analysis of randomized trials. AB - OBJECTIVE: To determine if luteal phase support improves the pregnancy rate in in vitro fertilization (IVF) cycles. DESIGN: A meta-analysis of randomized trials of luteal phase support was carried out with the main outcome measure being the pregnancy rate per cycle. RESULTS: Fifty-nine trials were evaluated. Eighteen trials met the inclusion criteria. Five main themes were identified: human chorionic gonadotropin (hCG) versus progesterone; progesterone versus progesterone and hCG; progesterone versus placebo; hCG versus placebo, and hCG versus progesterone versus no support. CONCLUSION: Luteal phase support is definitely indicated in IVF treatment cycles. This meta-analysis favored hCG above progesterone as luteal phase support with respect to pregnancy rates. Further prospective randomized trials are needed to determine a definite consensus with respect to the duration of luteal phase support in IVF cycles. PMID- 15785075 TI - Biological relevance of E-cadherin-catenin complex proteins in primary epithelial ovarian tumours. AB - This study analysed the biological relevance of E-cadherin, alpha-catenin, beta catenin and gamma-catenin immunoexpression pattern (reduced vs. preserved phenotype) in epithelial ovarian tumours. Immunohistochemistry was used to evaluate the expression of these proteins in 154 epithelial ovarian tumours, consisting of 17 benign, 33 borderline and 104 malignant tumours. In borderline tumours, the immunoexpression pattern of E-cadherin (p = 0.014) and alpha-catenin (p = 0.030) associated with histological type. In malignant tumours, the immunoexpression pattern of E-cadherin was related with histological type (p = 0.001). The immunoexpression pattern of beta-catenin associated with histological type and tumour differentiation (p = 0.005, p = 0.025, respectively). The preserved phenotype of E-cadherin was most frequently observed in mucinous tumours, whereas reduced E-cadherin was most frequently observed in serous tumours. The preserved phenotype of beta-catenin associated with endometrioid carcinomas, while reduced beta-catenin associated with poorly differentiated serous and clear cell carcinomas. Although the reduced phenotype was the most frequent immunoexpression observed for all proteins of the E-cadherin-catenin complex in epithelial ovarian tumours, only beta-catenin showed a significant difference between benign, borderline and malignant tumours (p = 0.045), since borderline and malignant tumours most frequently showed the reduced phenotype. The immunohistochemical profile of beta-catenin was shown to be of biological relevance: reduced beta-catenin was correlated with loss of tumour differentiation and serous carcinomas that are known to depict aggressive biological behaviour in epithelial ovarian tumours. PMID- 15785076 TI - Dilution method to refold bacterially expressed recombinant Der f 2 and Der p 2 to exhibit the secondary structure and histamine-releasing activity of natural allergens. AB - BACKGROUND: Structurally refolded recombinant forms of major house dust mite group 2 allergens, Der f 2 and Der p 2, expressed in Escherichia coli, were prepared by solubilizing the insoluble products with urea and subsequently dialyzing against buffer. In this study, we determined conditions for refolding the urea-denatured recombinant Der f 2 and Der p 2 by one-step dilution as an alternative to dialysis, which requires several steps of handling and much time and cost. METHODS: The insoluble bacterial product containing recombinant Der f 2 was solubilized with a buffer containing 8 M urea, and the solution was diluted to various urea concentrations. The refolding efficiency in each dilution was estimated from the height of the peak corresponding to the folded recombinant Der f 2 and that containing the aggregated form on anion exchange chromatography. The structure and allergenicity of the purified recombinant Der f 2 and Der p 2 refolded using the dilution method were analyzed based on circular dichroism and a basophil histamine-releasing assay, respectively. RESULTS: Although the refolding efficiency decreased as the urea concentration in the dilution increased, experimental conditions whereby the protein and urea concentrations in the dilution were less than 0.5 mg/ml and 0.8 M, respectively, achieved maximum refolding efficiency. The recombinant allergens prepared by the dilution method exhibited the secondary structure and histamine-releasing activity of natural allergens purified from mite culture. CONCLUSIONS: The dilution method established in this study is more convenient in terms of handling, time, and cost than the dialysis method and will be useful for large-scale production and for the preparation of numbers of mutants to analyze IgE epitopes. PMID- 15785077 TI - Cloning and Expression of the Olea europaea allergen Ole e 5, the pollen Cu/Zn superoxide dismutase. AB - BACKGROUND: Recombinant DNA technology does provide pure, well-defined and reproducible products to be used for clinical purposes, by cloning and expressing the cDNA of allergens present in a specific extract. Ole e 5 is a pollen allergen of Olea europaea with an IgE-binding frequency of about 35%, which has been identified as a superoxide dismutase (SOD). The aim of this study was to clone the cDNA of Ole e 5, to express Ole e 5 in Escherichia coli and to characterize its immunoreactivity. METHODS: cDNA of Ole e 5 was amplified by nested 3'-RACE PCR and cloned in pGEX vector 6P expression vector. After sequencing of some clones and homology analysis, the rOle e 5 was produced in an E. coli strain as a fusion protein with GST and purified. Then, the protein immunoreactivity was evaluated by patients' IgE binding (ELISA, ELISA inhibition, and immunoblotting) and by rabbit anti-rOle e 5 binding (immunoblotting and immunoblotting inhibition). RESULTS: The sequence analysis of Ole e 5 cDNA confirmed that Ole e 5 is a Cu/Zn SOD, with an identity from 90 to 80% with SOD from other species. rOle e 5 was recognized by IgE from 39% of olive pollen-allergic patients tested; moreover, this binding was inhibited by the olive pollen extract. An anti-rOle e 5 antiserum raised in rabbit strongly reacted with a natural component of about 16-kDa molecular weight present in the olive pollen extract; moreover, this binding was inhibited by the recombinant protein. CONCLUSIONS: Ole e 5 is the first Cu/Zn SOD identified as an allergen in a pollen source. Due to the widespread presence of this enzyme, rOle e 5 allergen, cloned and expressed in a complete form in E. coli, could represent a good tool to investigate the allergen cross-reactivity between O. europaea pollen and other allergenic sources, such as plant foods and other pollens. PMID- 15785078 TI - A recombinant precursor of the mustard allergen Sin a 1 retains the biochemical and immunological features of the heterodimeric native protein. AB - BACKGROUND: Mustard has been an important cause of food allergy of increasing incidence in the last years. Sin a 1, a storage 2S albumin, is the most relevant allergen from this spice. METHODS: Pichia pastoris has been used as host for the recombinant production of the precursor form of Sin a 1 (rproSin a 1). rproSin a 1 was purified to homogeneity by three chromatographic steps: gel filtration, anion exchange and reverse-phase HPLC. Molecular characterization was performed using Edman degradation, mass spectrometry, amino acid composition, and circular dichroism. Immunological properties were analyzed by immunoblotting, ELISA, and ELISA inhibition experiments. RESULTS: We overexpressed rproSin a 1 as a single polypeptide with both large and small chains linked by an internal processed fragment at high yield. The purified rproSin a 1 (>95%) was obtained as a monomeric and soluble protein. rproSin a 1 showed equivalent structural and immunological properties to natural heterodimeric Sin a 1 allergen. rproSin a 1 was recognized by 75% of the patients allergic to mustard. The inhibitory capacity of rproSin a 1 to the total allergenicity of mustard extracts varied from 13 to 83% in different patients, with a mean value of 54%. CONCLUSIONS: rproSin a 1 is a good candidate to replace natural allergen in diagnosis protocols of mustard allergy. P. pastoris has been demonstrated to be a suitable expression system for the production of allergenic derivates of Sin a 1 that could be used for immunotherapy purposes in future. PMID- 15785079 TI - Nondiffusional release of allergens from pollen grains of Artemisia vulgaris and Lilium longiflorum depends mainly on the type of the allergen. AB - BACKGROUND: Upon contact with a wet surface, mature pollen grains hydrate and release proteins including allergens. Knowledge of the release mechanism of allergens that are mainly localized intracellularly may allow the design of strategies for inhibition of allergen release and the consequent sensitization process. METHODS: An improved pollen chromatography was performed with Artemisia vulgaris and Lilium longiflorum pollen. Using three elution media of different pH, osmolality and salt concentration mimicking various types of wet surfaces, the time-dependent elution profiles of total protein, a cell wall-bound acid phosphatase activity (acPase), allergenic (profilin, Art v 1) and nonallergenic molecules (14-3-3 protein, actin) were monitored. RESULTS: The release kinetics of total protein and cell wall-bound acPase followed an exponential decrease in both pollen species indicating a diffusion-based protein release, whereas the elution profiles of profilin, Art v 1 and 14-3-3 protein showed nondiffusion characteristics. No general dependence on pH, osmolality or salt concentration of the elution media was observable in the elution profiles. Under the applied conditions, actin was not released indicating that the pollen grains remained intact during the elution. CONCLUSION: The elution profiles of pollen allergens indicated that substantial amounts of these proteins do not diffuse from the cell wall or are released from intracellular compartments during imbibitional leakage. Instead, a mechanism seems to operate that involves translocation from the pollen cytoplasm to the extracellular environment by crossing an intact plasma membrane. Such a mechanism would probably allow the use of pharmaceuticals for inhibition of allergen release. PMID- 15785080 TI - Airway closure after antigen challenge in cynomolgus monkeys: effect of the histamine H1 receptor antagonist, chlorpheniramine maleate. AB - BACKGROUND: Airway closure is frequently observed in human asthma. However, limited information exists on the factors that cause this condition. In this study, an allergic cynomolgus monkey model was used to characterize the condition of airway closure and assess the contribution of histamine H1 receptors to this response. METHODS: Oscillatory lung mechanics, arterial blood gases during ventilation on 100% O2 and functional residual capacity (FRC) assessed by helium dilution were measured before and then 10 min and 24 h after Ascaris aerosol challenge in 12 male Ascaris-sensitive cynomolgus monkeys. The monkeys were pretreated with intravenous saline or chlorpheniramine maleate (0.3 mg/kg) in a randomized crossover design. RESULTS: Ascaris challenge produced a large increase in airway resistance, an increase in lung tissue damping (G) that measures ventilation inhomogeneity in the lung, a reduction in arterial oxygen tension (PaO2) during ventilation on 100% O2 and a reduction in FRC. These effects were seen 10 min after the Ascaris challenge, but by 24 h, these parameters had returned close to the baseline values. Chlorpheniramine maleate (0.3 mg/kg, i.v.) produced a 12-fold shift in the histamine bronchoconstrictor dose-response curve. Pretreatment of monkeys with chlorpheniramine maleate (0.3 mg/kg, i.v.) attenuated the increase in airway resistance induced by Ascaris challenge, but had only a small effect on the increase in G and the reductions in PaO2 and FRC after antigen. CONCLUSIONS: These results demonstrate that airway closure occurs immediately after the antigen challenge in allergic cynomolgus monkeys and that histamine H1 receptors contribute very minimally to this response. PMID- 15785082 TI - Symptoms of asthma and the home environment. The ISAAC I and III cross-sectional surveys in Munster, Germany. AB - BACKGROUND: The indoor home environment has been shown to be associated with the presence of respiratory symptoms and atopic disease. METHODS: Two cross-sectional surveys (1995-2000) were conducted, using data from the ISAAC phase I and III surveys, collected in Munster, Germany (n = 6,996, response 81.8%). We analyzed the prevalence ratio (PR) for several indoor exposures and asthma-related outcomes in 6- to 7-year-old children, adjusting for potential confounders. RESULTS: Positive associations were observed regarding exposure to molds, environmental tobacco smoke, cooking with gas and space heating with fossil fuels, but most associations were not statistically significant. Surprisingly, presence of a carpet was negatively associated with most respiratory conditions. When restricting the analysis to participants without avoidance of a carpet due to a history of atopic disease, the protective associations disappeared. Mostly, present pet ownership did not show positive associations with respiratory symptoms. However, ownership at different times in life revealed positive associations, particularly in regard to birds owned in the first year of life (PR 1.51, 95% CI 1.11-2.06, for sleep disturbance due to wheeze and PR 1.28, 95% CI 1.01-1.63, for wheeze during the last 12 months, respectively). CONCLUSIONS: Only few indoor factors were positively associated with the studied outcomes. The change of associations in participants that reported allergy-related avoidance of carpets suggests that the observed effects are a result of asthmatics' changed behavior. The effect of allergy-related change in behavior and the results observed concerning the ownership of pets at different ages underline the need of establishing a precise temporal relationship between disease and exposure. PMID- 15785081 TI - Elevation of plasma soluble T cell costimulatory molecules CTLA-4, CD28 and CD80 in children with allergic asthma. AB - BACKGROUND: The surface expression of T cell costimulatory molecules CTLA-4 and CD28 and their counter-ligands, B7 molecules (CD80, CD86), is differentially induced for T cell activation and expansion in allergic asthma. However, the role of their soluble forms in plasma has not yet been elucidated. In this study, we investigated whether expression is altered and whether soluble costimulatory molecules are clinically relevant in asthmatic patients. METHODS: Plasma concentrations of soluble CTLA-4 (sCTLA-4), CD28, CD80 and CD86 in 51 children with chronic allergic asthma with or without inhaled corticosteroid treatment, and 22 sex- and age-matched control subjects were measured by enzyme-linked immunosorbent assay. Plasma total IgE concentration was measured using a microparticle immunoassay. RESULTS: Asthmatic patients had higher logarithmic plasma total IgE concentration (IgE(log)) than healthy subjects (p < 0.0001). In non-steroid-treated patients, plasma sCTLA-4, sCD28 and sCD80 but not sCD86 concentrations were significantly higher than those of control subjects (all p < 0.05). Plasma sCD80 and sCD86 but not sCTLA-4 and sCD28 concentrations correlated significantly with IgE(log) of all subjects (p < 0.05). There were also significant positive correlations between sCTLA-4 and sCD28 (p = 0.0007), and between sCD80 and sCD86 in all asthmatic patients (p = 0.001). CONCLUSIONS: Plasma sCTLA-4, sCD28 and sCD80 concentrations are elevated in allergic asthma. The increased expression of these soluble proteins may reflect the dysregulation of T cell activation, contributing to the immunopathogenesis of allergic asthma. PMID- 15785083 TI - Intentional delay of human hepatocarcinogenesis due to suppression of chronic hepatitis. AB - Human hepatocellular carcinomas (HCCs) are preceded by chronic hepatitis and cirrhosis. Despite a clear viral etiology [hepatitis B virus (HBV) and hepatitis C virus (HCV)] of human hepatocarcinogenesis, the mechanism is complex and the distinct molecular pathway or molecules to explain this phenomenon are not yet known. Hepatitis viral, 'inflammation-mediated' hepatocarcinogenesis greatly influences the incidence of somatic genetic events in hepatocytes by increasing the number of target cells, or the proliferation of once-hit hepatocytes, eventually leading to HCCs. These conditions may be designated as the 'hypercarcinogenic state'. Our goal is to lead the 'hypercarcinogenic state' to the 'normo- or hypocarcinogenic' state and to prevent HCC development (intentional delay of HCC). PMID- 15785084 TI - Single nucleotide polymorphisms associated with hepatocellular carcinoma in patients with chronic hepatitis B virus infection. AB - It is estimated that there are millions of single nucleotide polymorphisms (SNPs) within human genome and there are likely to explain much of the genetic diversity of individuals. Hepatocellular carcinoma (HCC) is etiologically associated with hepatitis B virus (HBV) in 80% of cases, and is the dominant cause of death among HBV carriers. Among patients with chronic HBV infection, family history is a known risk factor for the development of HCC; therefore, genetic factors are likely to modify the risk of HCC. However, the genetic factors that determine progression to HCC remain mostly to be investigated. In this review, we discussed that the natural history of HBV infection and host genetic factors related to HCC, study design and target gene selection for the detection of SNPs related to the occurrence of HCC. Also, we reviewed that several SNPs or haplotypes, which were reportedly associated with increased or reduced risk of HCC occurrence in patients with chronic HBV infection. Screening of these polymorphisms might be useful in clinical practice to stratify the lower or higher risk group for HCC and might modify the design of HCC surveillance programs in patients with chronic HBV infection, if further genetic susceptibilities are identified. PMID- 15785085 TI - Large liver cell dysplasia in hepatitis B virus x transgenic mouse liver and human chronic hepatitis B virus-infected liver. AB - OBJECTIVES: Large liver cell dysplasia (LCD) is frequently associated with hepatitis B virus (HBV), but it remains uncertain whether it is reactive, senescent or preneoplastic. METHODS: The HBX transgenic mice and normal control mice were sacrificed at 1, 3, 5, 7, 9, 11, 13 and 15 months after birth. Twenty three cases of human B viral chronic hepatitis/cirrhosis with prominent LCD were selected. The immunohistochemical stain of proliferating cell nuclear antigen (PCNA), transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and senescence-associated beta-galactosidase (SA-beta-Gal) were evaluated. RESULTS: In HBX transgenic mice, LCD was developed since 3 months and formed small nodules of hepatocellular adenoma, which progressed to hepatocellular carcinoma. The hepatocytes with LCD in HBX transgenic mice showed significantly higher PCNA labeling index (LI) and lower TUNEL-LI than normal hepatocytes of control mice (p < 0.05). In the majority of human B viral chronic hepatitis/cirrhosis, the hepatocytes with LCD revealed higher PCNA-LI and lower TUNEL-LI than those without, when compared in each case using the same tissue block. SA-beta-Gal staining showed no difference between hepatocytes with and without LCD. CONCLUSION: It is suggested that LCD, related to HBV, might not be just an innocent bystander, but closely related to hepatocarcinogenesis. PMID- 15785086 TI - How to predict HCC development in patients with chronic B viral liver disease? AB - Hepatocellular carcinoma (HCC) is one of the major malignant diseases in Asia. Although no randomized controlled studies have demonstrated decreased mortalities, screening for HCC has become an accepted procedure in the high-risk population. Since the incidence of HCC is strikingly different according to geographical distribution of risk factors, especially chronic hepatitis B virus (HBV) infection, a surveillance strategy in high endemic areas should be established according to national conditions. Chronic necro-inflammation by persistent HBV infection per se can progress to cirrhosis and the occurrence of HCC. As in other chronic liver diseases, the more risk factors the chronic B viral patients have, the higher occurrence of HCC was shown. Based on the risk factors of 4,339 Korean patients, the individual prediction model (IPM) was made by the calculation of relative weighs of risk factors and a self-exploited screening program for HCC was established accordingly. US screening at 6-month intervals was beneficial for the early detection of HCC, especially in the high risk group, and prolonged survival. The usefulness of a tailored screening program with IPM for HCC was prospectively confirmed. The effective screening system for diagnosis of HCC limited to patients with chronic hepatitis B might be necessitated in high endemic area. PMID- 15785087 TI - Hepatitis B virus-related hepatocellular carcinogenesis and its prevention. AB - To elucidate the influence of serum hepatitis B virus (HBV) load on hepatocellular carcinogenesis in cirrhotic patients, HBV-DNA was sequentially measured. In a nested, case-control study using 96 patients without antiviral therapy, high HBV-DNA (> or =10(3.7) copies/ml) in the last 3 years was significantly associated with carcinogenesis (a patient group without hepatocellular carcinoma (HCC) development; 0/48 vs. a patient group with eventual HCC development; 22/48, p < 0.0001). No patient with a continuously low HBV-DNA for the last 3 years developed HCC. Persistence of high HBV-DNA concentration suggested an increased risk of carcinogenesis. In a retrospective cohort study using 57 patients with interferon therapy, HCC developed in 2 (8.0%) of the 25 patients with HBV-DNA loss, while carcinogenesis was found in 11 (34.4%) of 32 patients without HBV-DNA loss (Fisher's exact test, p = 0.026). A significant decrease or loss of serum HBV-DNA stops HCC development, and its sequential analysis could be very useful both in the prediction and early detection of small HCC. PMID- 15785088 TI - Prevention of second primary tumors by an acyclic retinoid in patients with hepatocellular carcinoma. Updated analysis of the long-term follow-up data. AB - Oral administration with acyclic retinoid, a synthetic vitamin A analog, for a limited period of 12 months (48 weeks) prevented the development of second primary hepatocellular carcinoma (HCC) and also improved the survival of patients who underwent curative treatments of the initial tumor. Following that randomized controlled study reported in 1996 and 1999, we have continued to follow up the patients by medical imaging and blood chemical analyses, and found that the preventive effect of acyclic retinoid lasted up to 199 weeks after randomization (or 151 weeks after completion of retinoid administration). The retinoid's effect was not mediated by reduction in hepatic necro-inflammation since no significant decrease in serum aminotransferase activity was seen in the retinoid group. Such observation seems quite distinct from the cancer-preventive mechanism of interferon, a potent immunopreventive agent for HCC. We have also shown here the reduction by the retinoid in serum levels of lectin-reactive alpha-fetoprotein (AFP-L3) and protein induced by vitamin K absence or antagonist-II (PIVKA-II), both of which indicate the presence of latent HCC cells. These results suggest that acyclic retinoid may delete such malignant clones before they expand to clinically detectable tumors and thereby inhibited second primary HCC. Once such latent clones are eradicated, it may well take at least several years for the next cancer clone to arise clinically. This may possibly explain a reason for the long-term effect of the retinoid even after the limited period of administration. PMID- 15785089 TI - A strategy for early detection of recurrent hepatocellular carcinoma following initial remission by transcatheter arterial chemoembolization. AB - OBJECTIVES: To find a better surveillance method in detecting recurrent HCCs, patterns of recurrences following initial remission by transcatheter arterial chemoembolization (TACE) were evaluated. METHODS: Of 235 consecutive HCC patients who underwent TACE, 69 with initial remission were followed for >12 months. We compared the recurrence rates according to the characteristics of original HCCs and analyzed the locations of recurrent HCCs. We also evaluated the diagnostic efficacies of CT scan with serum AFP, angiography and Lipiodol CT scan in detecting recurrent HCCs. RESULTS: In 37 of 69, recurrent HCCs were detected after a median period of 17 months. Multinodular HCCs recurred more frequently than single-nodular HCCs. All of 5 patients with portal vein thrombosis recurred. Although 46% of recurrences were adjacent to original tumors, 62% were separated from them (8% at both). HCC with heterogeneous lipiodol uptake frequently recurred adjacent to original tumors. Only 18 of 37 recurrent HCCs were initially detected by serum AFP and CT scans; 17 by angiography, 2 only by lipiodol CT scan. CONCLUSIONS: Regular angiography may be valuable in detecting recurrent HCCs, especially in multinodular HCC. HCC with heterogeneous lipiodol uptake should be treated in combination with local ablation therapy. PMID- 15785090 TI - Prediction of efficacy of interferon treatment of chronic hepatitis C and occurrence of HCC after interferon treatment by a new classification. AB - One hundred and twenty-one chronic hepatitis C patients were administered interferon (IFN) and divided into two groups: 31 complete responders (CR) with prolonged HCV-RNA negative 1 year after treatment and 90 non-complete responders (non-CR), including partial responders (PR) with transiently negative HCV-RNA and non-responders (NR) with continuously positive HCV-RNA. Liver biopsy specimens were classified into four grades and stages according to the degree of severity and the extent of fibrosis, respectively. No correlation was observed between the rate of IFN efficacy and grading. By staging, however, a difference in the efficacy of IFN was observed between F1 or F2 and F3 (0.05 < p < 0.1 and 0.01 < p < 0.025, respectively). Of the CR, 0% (0/5) were at F0 and 27.9% (24/86) at F1, 42.9% (6/14) at F2, and 6.3% (1/16) at F3. Another group of 118 chronic hepatitis patients (31 CR, 41 PR and 46 NR) followed up for over 2 years after IFN treatment were analyzed. By staging, 7 cases were at F0, 76 at F1, 18 at F2, and 17 at F3. HCC occurred in 1 of the 31 CR cases (representing an annual incidence rate of 1.21%), in 4 of the 41 PR cases (4.08%), and in 4 of the 46 NR cases (3.55%). HCC did not occur in any of the 7 cases at F0 (representing an annual incidence rate of 0%); it occurred in 2 of the 76 cases at F1 (1.01%), in 1 of the 18 cases at F2 (2.28%), and in 6 of the 17 cases at F3 (16.57%). These results suggest that the new classification would be conducive to roughly predicting the efficacy of IFN treatment and the occurrence of HCC after IFN treatment. PMID- 15785091 TI - Development of hepatocellular carcinoma after interferon therapy in chronic hepatitis C. Is it possible to reduce the incidence by ribanirin and IFN combination therapy? AB - OBJECTIVES: Although the incidence of hepatocellular carcinoma (HCC) has been shown to be reduced after interferon (IFN) monotherapy in chronic hepatitis C, the risk factors for the development of HCC have not been fully understood. The aim of this study is to investigate the risk factors for the development of HCC after IFN in chronic hepatitis C as well as whether the incidence of HCC will be reduced by ribavirin and IFN combination therapy or not. METHODS: 495 patients with chronic hepatitis C and which received IFN monotherapy were followed and the incidence and risk factors for the development of HCC were examined. On the other hand, in the patients which received ribavirin and IFN combination therapy, the sustained response rate was assessed and the reduction rate of HCC development was predicted. RESULTS: Multivariate analysis by the Cox proportional hazard model revealed that the risk factors for HCC development were age, male gender, severe fibrosis and outcome of IFN therapy. On ribavirin and IFN combination therapy, the sustained response rate reached 17.3% in genotype 1b and 74% in genotypes 2a and 2b infection, thus reducing 20% of the estimated incidence of HCC. CONCLUSION: To reduce the incidence of HCC in chronic hepatitis C, improvement of the sustained response rate is an essential issue, and ribavirin and IFN combination therapy shows to be promising. PMID- 15785092 TI - Low-dose, long-term, intermittent interferon-alpha-2b therapy after radical treatment by radiofrequency ablation delays clinical recurrence in patients with hepatitis C virus-related hepatocellular carcinoma. AB - OBJECTIVE: To assess whether or not interferon (IFN) therapy prevents recurrence, and eventually improves the prognosis of patients with hepatocellular carcinoma (HCC) after completion of radical radiofrequency ablation (RFA) therapy. METHODS: Included as the IFN group in this study were 24 patients in total, who received radical RFA therapy first, followed by medication with IFN-alpha2b at such a low dose of 3 MIU x 2/week for as long as possible. On the other hand, the control group comprised 33 patients in total, who received radical RFA therapy without subsequent treatment with IFN. The control group was matched to the IFN group in age, platelet counts and size of nodules. RESULTS: Of the 24 patients treated with IFN, only one patient showed sustained virologic response. The median tumor free period until the first recurrence after radical RFA therapy was 3.4 years in the IFN group and 1.4 years in the control group (p = 0.02). During the first 3 years after commencement of IFN administration, the cumulative recurrence rate in the IFN group was found to be lower than in the control group (p = 0.01); however, with the lapse of time over 3 years, the recurrence rate in the IFN group increased. There was no difference in the cumulative survival rates between the IFN group and the control group (p = 0.25). CONCLUSION: Subsequently after radical RFA therapy, long-term, low-dose, intermittent IFN therapy successfully delayed clinical recurrence of HCC. PMID- 15785093 TI - Effect of long-term postoperative interferon therapy on intrahepatic recurrence and survival rate after resection of hepatitis C virus-related hepatocellular carcinoma. AB - OBJECTIVE: This study was aimed at evaluating the effects of interferon (IFN) alpha on survival rate after resection of hepatocellular carcinoma. METHODS: In a randomized, controlled trial by the University Hospital, Medical Center and affiliated hospital in Osaka, Japan, 30 men were after surgery randomly allocated to an IFN-alpha group (15 patients) and to a control group. Patients in the IFN group received 6 MIU of IFN-alpha intramuscularly daily for 2 weeks, then three times a week for 14 weeks, and finally twice a week for 88 weeks. The incidence of recurrence and survival rate were then studied. RESULTS: The response to IFN was sustained viral response (SVR) in 2 patients, biochemical response (BR) in 6, partial response (PR) in 5, and no response (NR) in 2. In the control, 8 of the 15 patients demonstrated continuous abnormally high levels of ALT. At the end point of the study, intrahepatic recurrence was detected in 9 of the IFN group and in 13 of the control (p = 0.065, log-rank test). The cumulative survival rate was higher in the IFN group than in the controls (p = 0.041). CONCLUSION: Postoperative IFN therapy improves the outcome after resection of hepatitis C virus-related hepatocellular carcinoma. PMID- 15785095 TI - BRCA1 gene expression in breast cancer in Kuwait: correlation with prognostic parameters. AB - OBJECTIVES: This study assessed the BRCA1 gene expression in breast cancer in Kuwait, and compared it with other known prognostic factors for the disease. MATERIALS AND METHODS: Forty-eight random samples of archival paraffin-embedded breast cancer tissues were studied for BRCA1 gene expression. Immunohistochemical method utilizing antibodies against different epitopes on the BRCA1 protein was used to study BRCA1 protein expression. In addition, for 29 patients, reverse transcription-polymerase chain reaction was used to detect BRCA1 mRNA expression. BRCA1 expression was correlated with age, histological type and grade of breast cancer, estrogen and progesterone receptor status, and C-erbB-2 expression. RESULTS: No demonstrable BRCA1 mRNA and protein expression was found in 79 and 83% of the breast cancer tissues, respectively. A positive relationship was demonstrated between lack of BRCA1 (mRNA and protein) expression and high histological grade, negative estrogen and progesterone receptor status, and overexpression of C-erbB-2 in the breast cancer tissues. CONCLUSIONS: The study demonstrated lack of BRCA1 gene expression (mRNA and protein) in the majority of breast cancers in Kuwait and confirmed the inverse relationship between BRCA1 expression and parameters that determine poor prognosis in breast cancer. PMID- 15785094 TI - Heparin regulates transcription of endothelin-1 gene in endothelial cells. AB - Heparin, which is widely used as an anticoagulant, has been shown to have antiatherosclerotic and antihypertensive effects in animals and humans. These effects are mediated by the inhibition of endothelin-1 (ET-1) production in endothelial cells. To clarify the mechanism of this inhibition, we investigated the effect of heparin on transcriptional regulation of the ET-1 gene in bovine aortic endothelial cells (BAEC) cultured in fetal calf serum. ET-1 mRNA expression was significantly suppressed by heparin in a dose-dependent manner. Promoter analysis revealed that the minimum ET-1 promoter containing only the GATA and AP-1 sequences as positive cis-acting sites in the ET-1 promoter is sufficient for this suppression. Gel mobility shift assays using oligonucleotides encoding the ET-1 AP-1 and ET-1 GATA sites confirmed that both AP-1 and GATA binding activities in BAEC nuclear extract were markedly inhibited by heparin. Western blot analyses indicated that heparin completely blocked extracellular signal-regulated kinase (ERK) activation, and inhibiting ERK activity resulted in loss of heparin-dependent inhibition of the ET-1 gene. These data indicate that the ET-1 mRNA level is negatively regulated by heparin at the transcription level, through modification of AP-1 and GATA protein binding activities, which direct the ET-1 promoter in BAEC. This effect may be mediated, at least in part, through inhibition of ERK activity. PMID- 15785096 TI - Distribution of apolipoprotein E alleles in the Omani population. AB - OBJECTIVE: To estimate the apolipoprotein E (apo E) allele distribution in the Omani population and to compare them with those of other populations. SUBJECTS AND METHODS: One hundred and sixty-two healthy Omanis of Arab Bedouin origin were genotyped by polymerase chain reaction followed by restriction fragment length polymorphism. RESULTS: The apo E allele frequencies were: epsilon2, 0.052; epsilon3, 0.886; epsilon4, 0.062. This pattern of distribution, characterized by the lowest epsilon4 and among the highest epsilon3 allele frequencies in the world, was very similar to that of Arabs, Southern Europeans of the Mediterranean basin, Indians, and Japanese populations. CONCLUSION: The results indicate that the allelic distribution of apo E in healthy Omanis is characterized by low Apo epsilon4 and high epsilon3 allele frequencies similar to those of other Arab, Southern European, Japanese and Indian populations. The homogeneous distribution of apo E alleles in this group of populations might have been influenced by diet and/or genetic admixture. PMID- 15785097 TI - Cross-reaction of sera from patients with various infectious diseases with Leishmania infantum. AB - OBJECTIVE: To compare and evaluate the application of indirect fluorescent antibody (IFA) and counterimmunoelectrophoresis (CIEP) for laboratory identification of visceral leishmaniasis. MATERIALS AND METHODS: Serum samples from patients with malaria (Plasmodium vivax, n = 86; Plasmodium falciparum, n = 38), brucellosis (n = 26), tuberculosis (n = 31) and typhoid fever (n = 35) were examined for the presence of antibody to Leishmaniainfantum antigen using IFA and CIEP tests. RESULTS: Using IFA, false-positive results were malaria (P. vivax 19.8%, P. falciparum 13.2%), tuberculosis (6.4%), brucellosis (3.8%), and typhoid fever (2.8%). Using CIEP, a lower percentage of false-positives was observed only among malaria patients (P. vivax 2.3%, P. falciparum 2.6%). Serum samples from patients with other infectious diseases were negative in the CIEP test. CONCLUSION: Based on the results of this study, the CIEP technique is recommended for immunodiagnosis of visceral leishmaniasis, especially in regions where malaria, brucellosis and tuberculosis are prevalent. PMID- 15785098 TI - Blood component transfusion in primary coronary artery bypass surgery in Kuwait. AB - OBJECTIVE: The aims of this study were to determine the rate of blood product transfusion, associated perioperative factors and cost of such blood product transfusion in primary coronary artery bypass surgery (CABG). SUBJECTS AND METHODS: The medical records of 159 consecutive primary CABG patients (142 male, 17 female) from January 1, 2003 to June 30, 2003 at Chest Diseases Hospital, Kuwait, were reviewed. Urgent and emergency cases were included. RESULTS: The mean age of the patients was 57.2 (range 36-77 years). Overall, 128 (80.5%) patients received blood product transfusion during primary CABG: 113 (70.5%) packed red blood cells (RBC), 54 (33.9%) fresh frozen plasma, and 13 (8%) platelets. Overall, 23 patients (12.6%) received more than two RBC transfusions intraoperatively. Significant factors for intraoperative RBC transfusion were: age >60 years, female gender, preoperative hemoglobin <12 g/dl, and 3 or more coronary bypass grafts constructed. One hundred and fifty-nine patients consumed 342 units of RBC at an average of 2.1 RBC units per patient. The cost per patient was 80 Kuwaiti dinar (KD; USD 240). CONCLUSION: The findings indicate a high rate of blood component transfusion in primary CABG patients in Kuwait that could expose the patients to the possible adverse effects, and such transfusions have high economic impact. PMID- 15785099 TI - Prevalence of type 1 diabetes among 6- to 18-year-old Kuwaiti children. AB - OBJECTIVES: To determine the prevalence of type 1 diabetes among 6- to 18-year old Kuwaiti children according to gender, age, and region. SUBJECTS AND METHODS: Children with type 1 diabetes aged 6-18 years were identified at 182 schools (50 primary, 63 intermediate, and 69 secondary) in Kuwait during the study period October 2000 to September 2002. Schools were randomly selected using the 2000/01 educational districts' registers as sampling frame proportional to the number of schools in each district. Prevalence rates were adjusted to the 2002 Kuwaiti population. Diagnosis of type 1 diabetes was based on the World Health Organization, and the American Diabetes Association criteria. RESULTS: Prevalence of type 1 diabetes was 269.9 per 100,000 (95% confidence interval, CI 241.6 298.3). There was no significant difference in prevalence between male (247.6, 95% CI 205.2-290.0) and female (285.5, 95% CI 247.5-323.5). Type 1 diabetes was more prevalent in the age group 10-13 years (347.3), and lowest in the age group 6-9 years (182.6) per 100,000; the difference was significant at p < 0.001. The overall age-adjusted prevalence rate was 252.9 (95% CI 234.6-271.2), 229.1 (95% CI 204.6-253.6) in male and 277.4 (95% CI 250.0-304.7) in female children in the 2002 Kuwaiti population. The mean age at onset was 9.2, and 8.1 years in male and female children, respectively (p = 0.018). There was no significant difference in prevalence between regions. CONCLUSION: Type 1 diabetes is a common chronic disease in Kuwaiti children. PMID- 15785100 TI - Reliability of Omani women's self-reporting of gynaecologic morbidities. AB - OBJECTIVES: To assess the agreement of women's self-reports of gynaecologic morbidities (reproductive tract infections, RTIs; genital prolapse, and urinary tract infections, UTIs), physician's observations and the final diagnosis of these conditions. SUBJECTS AND METHODS: A nationally representative sample of 1,662 Omani women selected by a multi-stage, stratified probability-sampling procedure was included in the study. A questionnaire interview, physical and gynaecologic examination, and laboratory investigations were used. RESULTS: A total of 1,364 women completed all forms. The agreement between women's report of vaginal discharge and the diagnosis of RTI was poor. The sensitivity but not the specificity improved when the women were asked about specific discharge questions that upset them or are medically suspicious. Sensitivity of self-report in the diagnosis of genital prolapse and UTI from self-report was high but specificity was low. CONCLUSIONS: Self-report of vaginal discharge, coupled with clinical examination, can be used in community diagnosis of RTI. Self-report of symptoms of vaginal prolapse can also be used for community diagnosis of genital prolapse. A urinary culture should supplement self-reports of UTI to enhance specificity, although the presence of a positive culture in the absence of symptoms is of no clinical significance in non-pregnant women or women without renal disease. PMID- 15785101 TI - Double-catheter technique for the proper insertion of umbilical venous catheters in newborns. AB - OBJECTIVES: To document the usefulness and safety of inserting a second umbilical venous catheter in ill neonates, while a previously misplaced first catheter was still in its place. SUBJECTS AND METHODS: The case series involved 25 newborn babies who were admitted to the Neonatal Intensive Care Unit, Maternity Hospital, Kuwait, over a 3-year period from 1999 to 2002. The umbilical venous catheter of the babies was misplaced and diverted to the liver, necessitating insertion of a second catheter while the previous one was still in place. The characteristics of the babies and possible catheter-related complications were recorded. RESULTS: Of the 25 babies, 19 had the second catheter properly placed in the right atrium, while in the remaining 6 neonates, the catheter was still misplaced. Misplacement occurred mostly in full-term babies or the catheter was inserted at a later stage. No life-threatening complication was observed during the procedure. CONCLUSION: Insertion of a second umbilical venous catheter with the misplaced first catheter in situ is a useful and safe procedure. PMID- 15785102 TI - Prevalence and genotypes of hepatitis G virus among hemodialysis patients in Eastern Anatolia, Turkey. AB - OBJECTIVES: To study the prevalence and genotype distribution of hepatitis G virus (HGV) in hemodialysis patients in East Anatolia, Turkey. SUBJECTS AND MATERIALS: Eighty-nine hemodialysis patients and 30 healthy individuals were analyzed by using reverse-transcriptase nested polymerase chain reaction with primers specific for 5' untranslated region. HGV genotyping was performed by PCR and three randomly selected HGV-positive samples were sequenced. RESULTS: Of the 89 hemodialysis patients, HGV RNA was detected in 9 (10.2%). All of our isolates were assigned to genotype 2. CONCLUSION: Our results showed that hemodialysis patients carry the risk for HGV infection in East Anatolia, Turkey. PMID- 15785103 TI - Contrast media extravasation: manual versus power injector. AB - OBJECTIVE: To compare the incidence and complications of extravasation of low osmolar nonionic contrast media, injected manually and by the automatic power injector (API). SUBJECTS AND METHODS: Three thousand five hundred and sixty patients underwent contrast-enhanced abdominal and thoracic computerized tomography scan in the Department of Clinical Radiology, Al-Amiri Hospital, Kuwait, between June 1998 and December 2002. These patients were prospectively analyzed for contrast media extravasation, its relation to injection rate, cannula insertion and gauge and its complications. 920 patients were administered low-osmolar nonionic contrast media (Ultravist 300, Omni Paque 240 or 300) intravenously by manual injection and 2,640 patients by automatic power injector. RESULTS: Of the 3,560 patients contrast media extravasation occurred in 11 (0.3%). The symptoms were observed in 9 patients (0.3%) in the API group and 2 patients (0.2%) in the manual injection group, respectively. None of the patients had any soft tissue injury. CONCLUSION: The incidence of contrast media extravasation is not significantly increased by the use of the API. Low-osmolar nonionic contrast media extravasation resulting from the use of API does not cause any morbidity. PMID- 15785104 TI - Renography within 1 hour of renal transplantation: technical feasibility and clinical utility. AB - OBJECTIVE: Renographies obtained within 1 h of renal transplantation were studied prospectively to evaluate their technical feasibility and potential clinical impact on successful treatment of immediate posttransplantation complications such as arterial or venous thrombosis, which require prompt diagnosis and management. SUBJECTS AND METHODS: During December 1996 to December 1998, 127 renal transplants were performed. Ninety-four patients had complete renographic studies within 1 h of surgery. It was not possible to perform renography on 26 patients who were not sent to the Department of Nuclear Medicine within 1 h of transplant and 7 other patients who could not undergo a complete renogram because of their postoperative condition. RESULTS: There was no complication attributed to transporting patients to the Department of Nuclear Medicine for these studies so soon after surgery. Of the 94 renographies obtained immediately following transplant surgery 46 were abnormal. However, when compared with the usual policy of obtaining baseline renograms between 12 and 72 h after surgery, treatment was changed for only 2 patients. Radionuclide renography within 1 h of transplant surgery was technically feasible. CONCLUSION: Based on the results of this study the clinical utility of obtaining renography within 1 h posttransplant was minimal and hence we recommended that it should not be performed routinely but could be used on an individual basis when imminent intervention is highly likely. PMID- 15785105 TI - Glucocorticoid replacement in panhypopituitarism complicated by myelinolysis. AB - OBJECTIVE: To report a case of glucocorticoid substitution in panhypopituitarism that can lead to uncontrolled rise in serum sodium and myelinolysis. CLINICAL PRESENTATION AND INTERVENTION: A 42-year-old man presented with disturbed conscious level and hyponatremia. Initial data suggested glucocorticoid deficiency. Later, hormonal levels indicated panhypopituitarism. MRI of the brain led to the diagnosis of a pituitary macroadenoma. Glucocorticoid substitution was initiated immediately after admission, and possible myelinolysis subsequently became a complication. We report this case to illustrate the fact that glucocorticoid substitution can lead to rapid rise in serum sodium and myelinolysis in panhypopituitarism. CONCLUSION: This case illustrated the need to use minimum doses of glucocortcoids with close monitoring of serum sodium, in order to avoid this complication. PMID- 15785106 TI - Brucellosis: haemorrhagic pleural effusion. AB - OBJECTIVES: To describe haemorrhagic pleural effusion as a rare complication of brucellosis that finally needed lung decortication. CLINICAL PRESENTATION AND INTERVENTION: A 37-year-old female presented with a 1-week history of fever, dry cough and pleuritic chest pain. Physical examination showed signs of right pleural effusion and hepatosplenomegaly. Complete blood count showed pancytopenia, white blood cells 2.9/mm3, haemoglobin 10 g/dl, platelets 131/mm3. Chest X-ray confirmed a moderate right pleural effusion, that was found to be exudative biochemically. Culture of pleural fluid and blood grew Brucella species. Fever subsided with Brucella chemotherapy, but pleural effusion persisted. Computed tomographic (CT) chest scan showed a large loculated pleural effusion, which failed to resolve despite repeated aspirations under CT guidance. Fluid was always found to be haemorrhagic. Finally, lung decortication was done with successful outcome. CONCLUSION: This case showed that brucellosis can cause haemorrhagic pleural effusion that needs lung decortication. PMID- 15785107 TI - Diverticular carcinoma of the urinary bladder: diagnosis and treatment problems. A case report. AB - OBJECTIVES: To report a case of a primary carcinoma arising in a vesical diverticulum. CLINICAL PRESENTATION AND INTERVENTION: A 59-year-old male patient presented with painless macrohematuria. A primary carcinoma arising in a vesical diverticulum was detected with cystoscopy and confirmed with rectal endosonography and computer tomography. and radical cystoprostatovesiculectomy with ileal conduit was performed. Chemotherapy with cisplatin and methotrexate followed. The patient died of an acute cardiac event in the 4th postoperative month. CONCLUSION: This report illustrates that in a case of a closed opening of a bladder diverticulum, rectal endosonography and computer tomography do provide additional support for making a diagnosis of a hidden tumor in the diverticulum. PMID- 15785108 TI - Complement component c5a is integral to the febrile response of mice to lipopolysaccharide. AB - OBJECTIVES: The complement system is critical to the febrile response of mice to intraperitoneally administered lipopolysaccharide (LPS). We previously identified C3 and C5 as two components potentially involved in this response. This study was designed to examine whether the complement system is also pivotal in the response of mice to intravenously or intracerebroventricularly injected LPS, to distinguish between C3 and C5 and their cognate derivatives as the essential mediator(s), and to determine whether the failure of complement-deficient mice to develop a fever could be due to their possible inability to secrete pyrogenic cytokines. METHODS: Wild-type (WT; C57BL/6J) mice, hypocomplemented or not by intravenously injected cobra venom factor (10 U/mouse), and C3-, CR3- and C5 sufficient and -deficient mice were intravenously challenged with LPS (0.25 mug/mouse); WT and C3-/- mice pretreated with a C5a receptor antagonist (C5aRa) were similarly challenged. In addition, the serum levels of interleukin (IL) 1beta, tumor necrosis factor (TNF)-alpha and IL-6 were compared in LPS-treated C5+/+ and C5-/- mice. RESULTS: LPS induced a 1 degrees C rise in core temperature in all the mice, except C5-/- mice and those pretreated with C5aRa. C5+/+ and C5 /- mice challenged intracerebroventricularly with LPS exhibited identical febrile responses. LPS induced similar increases in the serum levels of IL-1beta, TNFalpha and IL-6 in C5+/+ and C5-/- mice. CONCLUSIONS: C5a is crucial for the development of febrile responses to LPS in mice; its site of action is peripheral, not central. The possibility that an inability to produce cytokines could account for the failure of C5-/- mice to develop a fever is not supported. PMID- 15785109 TI - Increased dopamine content in lymphocytes from high-dose L-Dopa-treated Parkinson's disease patients. AB - OBJECTIVES: The intracellular (i.c.) content of dopamine and its metabolites was measured in the peripheral blood lymphocytes (PBLs) of Parkinson's disease (PD) patients and healthy controls. METHODS: Catecholamine levels of PBLs were measured using capillary electrophoresis in healthy controls and PD patients receiving different doses of L-dihydroxyphenylalanine (L-Dopa). RESULTS: Higher i.c. dopamine content was found in lymphocytes from PD patients receiving a high dose of L-Dopa (700 +/- 30 mg/day) as compared to lymphocytes from the healthy controls (p = 0.002) and from PD patients treated with a low dose of L-Dopa (400 +/- 30 mg/day) (p = 0.022). The dihydroxyphenylacetic acid to dopamine ratio was significantly lower in the high-dose L-Dopa-treated PD patients than in the controls (p = 0.013). CONCLUSIONS: These findings suggest that the dopamine content and metabolism in the peripheral lymphocytes of PD patients are influenced by L-Dopa administration. This is the first study in which a dose related effect of L-Dopa treatment was found in lymphocytes from PD patients. PMID- 15785110 TI - Hypothalamic control of mitogen-induced proliferative responses and luteinizing hormone-releasing hormone levels in thymus and peripheral blood of rat fetuses. AB - The role of endogenous luteinizing hormone-releasing hormone (LHRH) in the development of concanavalin A (ConA)-induced proliferative responses was studied in rat fetuses. Preliminary treatment of fetuses in utero with either the LHRH receptor antagonist or anti-LHRH antibodies resulted in the suppression of ConA induced proliferative responses of thymocytes. LHRH and LHRH-immunopositive cells, morphologically similar to thymocytes, were detected in intact fetal thymus. A significant content of LHRH was also found in the peripheral blood of fetuses. The LHRH content in thymus and plasma was similar in males and females. Surgical ablation of the hypothalamus resulted in 2-fold decreases in thymus and plasma levels of LHRH in 21-day-old fetuses compared to sham-operated fetuses. It was concluded that LHRH regulates mitogen-induced proliferative responses of thymocytes during prenatal ontogenesis in the rat. The main source of plasma LHRH at that period is the hypothalamus. Moreover, LHRH is synthesized in the fetal thymus. Thus, LHRH is suggested to have not only a central effect but also to be involved in autocrine or paracrine regulation of proliferative immune responses. PMID- 15785111 TI - Different mitogen-mediated Beta-adrenergic receptor modulation in murine T lymphocytes depending on the thyroid status. AB - OBJECTIVE: The aim of this work was to analyze beta-adrenergic receptor (betaAR) regulation of T-lymphocyte proliferation in mice according to different thyroid hormone statuses. METHODS: T cells from eu-, hypo- (by propylthiouracil treatment) and hyperthyroid (by thyroxine, T4 administration) mice were purified and specific radioligand binding assays were performed. The effects of the beta agonist isoproterenol (ISO) on intracellular levels of cyclic AMP (cAMP) were determined. Mitogen-induced T-cell proliferation was measured by [(3)H]-thymidine incorporation. Finally, protein kinase C (PKC) activity in cytosol and membrane fractions were determined using radiolabelled enzymatic substrates. RESULTS: Adecrease or a non-significant increase in betaAR number was found on T lymphocytes from hypo- and hyperthyroid mice compared to euthyroid controls. ISO stimulation of cAMP levels was lower in hypothyroid and higher in hyperthyroid T lymphocytes compared to controls. T-selective mitogen-induced proliferation was increased in T4-treated animals, but decreased in hypothyroid mice. During the peak of proliferation, downregulation of betaAR was observed in all animals. However, a higher or a lower decrease was observed in hyper- and hypothyroid T cells, respectively. In parallel, a higher translocation of PKC activity was observed in hyperthyroid cells, and a lower one was found in hypothyroid lymphocytes with respect to controls. CONCLUSIONS: These results indicate that intracellular signals triggered by mitogen activation, namely PKC, would be related to differential betaAR downregulation in T lymphocytes depending on the thyroid hormone status, contributing to the distinct proliferative responses found in hypo- or hyperthyroidism compared to the euthyroid state. PMID- 15785112 TI - Abnormal thymic microenvironment in insulin-like growth factor-II transgenic mice. AB - OBJECTIVES: Intrathymic T cell differentiation is driven by the thymic microenvironment, a tridimensional network of cells and extracellular matrix (ECM). Previous data showed that lymphoid and microenvironmental compartments are under the control of hormones and growth factors. We then attempted to define if insulin-like growth factor-II (IGF-II) was also involved in such a control. METHODS: We used IGF-II transgenic (Tg) mice and studied their thymic microenvironment by immunohistochemistry. Moreover, we evaluated thymocytes in terms of their ability to adhere to thymic epithelial cells and to migrate through epithelial cells and ECM. RESULTS: Transgenic IGF-II expression results in abnormalities of the thymic epithelium. Terminal differentiation of thymic epithelial cells (TEC) is modified, with the appearance of large clusters of cells immunoreactive to the monoclonal antibody KL1, which specifically recognizes highly differentiated TEC. Accordingly, treatment of cultured TEC with exogenous IGF-II induces the appearance of KL1+ cells and increases TEC proliferation. IGF-II Tg animals exhibit increased serum levels of the TEC derived hormone thymulin. These effects were seen even when the IGF-II transgene was inserted in dwarf mice. Moreover, deposition of fibronectin and laminin is also enhanced in IGF-II Tg mouse thymus and in IGF-II-treated TEC cultures. Furthermore, ECM-mediated interactions between thymocytes and TEC are affected by exogenous IGF-II, as exemplified by the enhancement of thymocyte adhesion to TEC monolayers and thymocyte migration in thymic nurse cell complexes. CONCLUSIONS: Our data indicate that IGF-II pleiotropically affects the thymic epithelium, both in vivo and in vitro, and that some of these changes may have consequences on thymocyte/TEC interactions. PMID- 15785113 TI - Evidence of an anti-inflammatory role for Vasogen's immune modulation therapy. AB - We have reported that Vasogen's immune modulation therapy (IMT), a procedure involving intramuscular administration of autologous/syngeneic blood, which has been exposed ex vivo to increased temperature, UVC light and oxidation, prevents several LPS-induced inflammatory changes in the hippocampus. Here, we investigated neuroprotective effects of IMT in cortical tissue, and report that the treatment acts as an anti-inflammatory and antioxidative agent, reducing the concentration of TNFalpha and the accumulation of reactive oxygen species. The data couple these changes with an increase in the concentration of the anti inflammatory cytokine IL-10, and a decrease in activation of the stress-activated protein kinase, c-jun N-terminal kinase. Consistent with these putative protective effects of IMT, we report that the LPS-induced increase in TUNEL staining, which is indicative of cell death, is prevented by IMT. PMID- 15785114 TI - Neonatal androgenization affects the intrathymic T-cell maturation in rats. AB - The thymus structure, expression of CD4/CD8/TCRalphabeta on thymocytes and thymocyte proliferative and apoptotic indexes were analyzed in sexually immature 30-day-old and in sexually mature 60-day-old female rats neonatally androgenized (NA) by subcutaneous injection of 500 microg testosterone propionate/day on days 1-3 and in their vehicle-administered counterparts. The treatment affected normal thymus development. Thus, at 30 days of age, there was a reduction in the thymus weight, reflecting a decrease in the main thymic compartments. However, at 60 days of age, thymus weight did not significantly differ from that in age-matched controls, since the cortical volume enlargement was followed by a proportional decrease in the medullary volume. In rats of both ages, the changes in thymic compartments most likely reflected alterations in the size of both lymphoid and nonlymphoid components. Furthermore, in NA rats, substantial changes in thymocyte phenotypic characteristics were registered, in spite of their age. In both groups of NA rats, a decrease in the relative proportion of the least mature CD4-8 TCRalphabeta- cells and in that of CD4+8- TCRalphabeta-/TCRalphabeta(low) cells followed by an increase in the percentage of their successor CD4+8+TCRalphabeta /TCRalphabeta(low) cells was detected. In addition, in 30-day-old NA rats, the relative proportions of CD4+8+TCRalphabeta(high) cells (just positively selected) and that of mature single positive (CD4+8- and CD4-8+) and CD4-8- double negative TCRalphabeta(high) cells, were reduced, while in 60-day-old NA rats only the percentage of CD4+8+TCRalphabeta(high) thymocytes was decreased. Thus, the study showed that the changes in the development of the hypothalamo-pituitary-gonadal axis induced by neonatal androgenization may affect the thymus development and intrathymic T-cell maturation. PMID- 15785115 TI - Using remifentanil and sufentanil in functional endoscopic sinus surgery to improve surgical conditions. AB - The purpose of the study was to compare three types of general anesthesia for functional endoscopic sinus surgery (ESS) with controlled hypotension measuring the quality of visibility of the surgical field and the blood loss during the operation. Seventy-one patients underwent endoscopic ethmoidectomy bilaterally for nasal polyposis and/or chronic sinusitis. The patients were divided into three groups according to the type of anesthesia they had: group A (sufentanil/sevoflurane), group B (remifentanil/propofol), and group C (fentanyl/isoflurane). The mean estimated blood loss for group A was 117.83 ml, for group B it was 100.5 ml and for group C it was 198.89 ml. The average quality of visibility of the surgical field was 1.57 for group A, 1.3 for group B and 2.79 for group C. The quantity of blood loss (p < 0.01) and the visibility of the surgical field (p < 0.001) demonstrated a difference among the three groups. Remifentanil and sufentanil during functional ESS enable controlled hypotension and a general improvement in surgical conditions. PMID- 15785117 TI - Expression of pituitary adenylate cyclase activating polypeptide and its type I receptor mRNAs in human placenta. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) was first isolated from ovine hypothalamus and was known to stimulate the release of growth factor in various cells. Recently, we reported the cellular localization of PACAP and its type I (PAC1) receptor in rat placenta during pregnancy. Placenta is a critical organ that synthesizes several growth factors and angiogenic factors for the fetal development and its own growth. However, there is little information regarding the cellular localization of PACAP and its receptor in human placenta at various gestations. The aim of the present study was to define the expression and distribution of PACAP and PAC1 receptor mRNAs in the human placenta during the pregnancy period. PACAP and PAC1 receptor mRNAs were expressed in stroma cells of stem villi and terminal villi. At the early stage, on 7 and 14 weeks, PACAP and PAC1 receptor genes were moderately expressed in stroma cells surrounding the blood vessels within stem villi. These genes were strongly expressed in stroma cells of stem villi and terminal villi on 24 and 38 weeks. The expression of these genes was increased as gestation advanced, and localized in the same areas. Localization of PACAP and PAC1 receptor demonstrate the evidence that PACAP may play an important role, as an autoregulator or pararegulator via its PAC1 receptor. In conclusion, our findings strongly suggest that PACAP may have a critical role in physiological function of the placenta for gestational maintenance and fetal growth. PMID- 15785116 TI - Evidence for the presence of beta-3-adrenoceptors mediating relaxation in the human oviduct. AB - Beta1- and beta2-adrenoceptors mediate relaxation in the oviductal smooth muscle. This study examines the existence and function of beta3-adrenoceptors in the human oviduct. Ring segments of the oviduct were set up for isometric tension recording. The effect of isoprenaline and BRL 37344 on smooth muscle tone was examined. The expression of beta3-adrenoceptors in the oviduct was also examined. Isoprenaline and BRL 37344 concentration-dependently relaxed circular muscles of the oviduct. BRL 37344 was less potent than isoprenaline and was a partial agonist. Propranolol shifted isoprenaline but not BRL 37344 concentration response curve to the right without reducing the maximum response. Cyanopindolol (1 micromol/l), a beta3-adrenoceptor antagonist, shifted the isoprenaline concentration response curve to the right. The -log K(B) value of 7.8 indicates activation of beta3-adrenoceptors by isoprenaline. mRNA for beta3-adrenoceptors was expressed in the oviduct. These results suggest that beta3-adrenoceptors, mediating relaxation, are expressed in the human oviduct. PMID- 15785118 TI - Isolation and identification of Escherichia coli O157:H7 using different detection methods and molecular determination by multiplex PCR and RAPD. AB - Escherichia coli O157:H7 is recognized as a significant food-borne pathogen, so rapid identification is important for food hygiene management and prompt epidemiological investigations. The limited prevalence data on Shiga toxin producing E. coli (STEC) and E. coli O157:H7 in foods and animals in Korea made an assessment of the risks difficult, and the options for management and control unclear. The prevalence of the organisms was examined by newly developed kit-E. coli O157:H7 Rapid kit. For the isolation of E. coli O157:H7, conventional culture, immunomagnetic separation, and E. coli O157:H7 Rapid kit were applied, and multiplex PCR and randomly amplified polymorphic DNA (RAPD) were performed for the molecular determination. There was high molecular relatedness among 11 Korean isolates and 17 U.S. strains at 63% level. Additionally, distinct differentiation between pig and cattle isolates was determined. It implied that RAPD had a capacity to distinguish strains with different sources, however it could not discriminate among isolates according to their differences in the degree of virulence. In antimicrobial susceptibility tests, 45.5% of isolates showed antibiotic resistance to two or more antibiotics. Unlike the isolates from other countries, domestic isolates of E. coli O157:H7 was mainly resistant to ampicillin and tetracyclines. In summary, the application of E. coli O157:H7 Rapid kit may be useful to detect E. coli O157:H7 due to its sensitivity and convenience. Moreover, combinational analysis of multiplex PCR together with RAPD can aid to survey the characteristics of isolates. PMID- 15785119 TI - Induction of castration by immunization of male dogs with recombinant gonadotropin-releasing hormone (GnRH)-canine distemper virus (CDV) T helper cell epitope p35. AB - Immunocastration is a considerable alternative to a surgical castration method especially in male animal species for alleviating unwanted male behaviors and characteristics. Induction of high titer of antibody specific for gonadotropin releasing hormone (GnRH) correlates with the regression of testes. Fusion proteins composed of canine GnRH and T helper (Th) cell epitope p35 originated from canine distemper virus (CDV) F protein and goat rotavirus VP6 protein were produced in E. coli. When these fusion proteins were injected to male dogs which were previously immunized with CDV vaccine, the fusion protein of GnRH-CDV Th cell epitope p35 induced much higher antibody than that of GnRH-rotavirus VP6 protein or GnRH alone. The degeneration of spermatogenesis was also verified in the male dogs immunized with the fusion protein of GnRH-CDV Th cell epitope p35. These results indicate that canine GnRH conjugated to CDV Th cell epitope p35 acted as a strong immunogen and the antibody to GnRH specifically neutralized GnRH in the testes. This study also implies a potential application of GnRH-based vaccines for immunocastration of male pets. PMID- 15785120 TI - Influences of DTC and zinc supplementation on the cellular response restoration in restrained mice. AB - The studies were conducted on Balb/c mice exposed to restraint stress twice for 12 h at 24 h intervals. Prior to restraint stress the mice were treated with sodium diethyldithiocarbamate (DTC) i.p. at a dose of 20 mg/kg five times at 48 h intervals. DTC was used per se or with zinc ions interaction, by adding zinc sulfate to drinking water at a dose of 72 microgram/mouse daily. The results obtained in the study show that restraint stress causes involution of lymphatic organs, decreased the percentage of immature (CD4+CD8+) and, mature (CD4+) thymocytes and CD4+), CD8+ and CD19+ splenocytes and proliferative response of thymocytes stimulated in vitro with concanavalin A (Con A) and phytohemagglutinin (PHA). The restraint stress decreased also interleukin-1 (IL-1) production by murine intraperitoneal macrophages stimulated in vitro with lipopolysaccharide (LPS) from E. coli. Pretreatment with DTC counteracted restraint stress-induced immunosuppression, which is expressed as partial normalisation of the total number of thymocytes, splenocytes and IL-1 production, accelerated regeneration of thymus and spleen, shorter suppressive action of restraint stress on the percentage of CD4+CD8+thymocytes and in total normalisation of the CD4+thymocytes and splenocytes. DTC administered prior to restraint stress augmented the proliferative response of thymocytes to two mitogens. The immunocorrecting action of DTC is enhanced by zinc supplementation, expressed in the increased percentage of CD4+thymocytes and splenocytes, CD19+splenocytes, proliferative activity of thymocytes stimulated with PHA and IL-1 production. The obtained results show that DTC administration can be supplemented with zinc in order to restore the immune system impaired by stress. PMID- 15785121 TI - Growth performance and nutrient utilization in black Bengal bucks (Capra hircus) supplemented with graded doses of chromium as chromium chloride hexahydrate. AB - Black Bengal bucks (Capra hircus) were supplemented with 0, 0.2 or 0.4 mg elemental chromium (Cr) as chromium chloride hexahydrate per day for 70 days. Intake of dry matter (p<0.001), crude protein (p<0.001) and neutral detergent fiber (p<0.01) increased due to Cr supplementation. The apparent total tract digestibility of dry matter (p<0.01), organic matter (p<0.05), crude protein (p<0.001) and acid detergent fiber (p<0.01) improved and the total body weight gain and the live weight gain to feed intake ratio also increased (p<0.001) due to supplemental Cr feeding. The intake and apparent absorption (p<0.001) of Cr was enhanced due to its supplementation. The intake of copper, zinc, manganese and iron was also more (p<0.001) in the Cr supplemented bucks. As supplementation progressed, plasma glucose concentration was elevated particularly in 0.4 mg Cr supplemented bucks and a significant day x dose interaction effect (p<0.001) with this parameter. The activity of plasma alkaline phosphatase increased (p<0.001) and that of glutamate pyruvate transaminase in plasma decreased (p<0.01) in the Cr supplemented bucks. Supplemental Cr had minimal (p>0.05) effect on the plasma half life (k) and clearance rate of glucose (T1/2) during an intravenous glucose tolerance test. Area under the response curve from 0 to 180 minutes after glucose loading was lower (p<0.001) in the control group of bucks. The study revealed that Cr supplementation might promote growth and nutrient utilization in black Bengal bucks. However, little difference between the 0.2 and 0.4 mg Cr supplemented bucks suggested limited benefit of increasing the level of supplementation beyond 0.2 mg per day under the normal management regimes. PMID- 15785122 TI - Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies. AB - Five monoclonal antibodies (MAbs) and chicken immunoglobulin (IgY) were developed by immunizing with flagella purified from Listeria monocytogenes 4b and the five MAbs have been confirmed to be specific against three different epitopes of flagellin. The antibodies showed specific reaction to Listeria genus and no cross reactivity with other bacteria tested in this experiment including E.coli O157:H7 and Salmonella enteritidis. Sandwich enzyme-linked immunosorbent assays (ELISA) using the MAbs and IgY were developed to detect Listeria species and the sensitivity and specificity of the developed ELISA have been analyzed. The detection limit of ELISA using MAb 2B1 and HRP labeled IgY was 1 x 10(5) cells/0.1 ml at 22 degrees C. and 1 x 10(6) cells/0.1 ml at 30 degrees C. ELISA using the pair of MAbs (MAbs 2B1 and HRP labeled MAbs 7A3) detected up to 10(4) cells/0.1 ml at 22 degrees C and 30 degrees C. Detection limit of sandwich ELISA using IgY was 10 times lower than MAb pair. Using the developed ELISA, we could detect several Listeria contaminated in food samples after 48 h-culturing. In conclusion, both MAbs and IgY have been proved to be highly specific to detect Listeria flagella and the developed sandwich ELISA using these antibodies would be useful tool for screening Listeria spp. in food. PMID- 15785123 TI - Immunization of mice with recombinant P27/30 protein confers protection against hard tick Haemaphysalis longicornis (Acari: Ixodidae) infestation. AB - The success of immunological control methods is dependent upon the use of potential key antigens as tick vaccine candidates. Previously, we cloned a gene encoding 27 kDa and 30 kDa proteins (P27/30) of Haemaphysalis longicornis, and identified the P27/30 is a troponin I-like protein. In this study, the recombinant P27/30 (rP27/30) expressed in Escherichia coli was used to immunize mice and the mice were challenge-infested with ticks at different developmental stages of the same species. The rP27/30 protein stimulated a specific protective anti-tick immune response in mice, evidenced by the statistically significant longer pre-feeding periods in adult ticks. Furthermore, significantly longer feeding periods were noted in both larval and adult ticks. On the other hand, only larval ticks exhibited low attachment rates (31.1%). Immunization of mice with rP27/30 protein confers protection against hard tick Haemaphysalis longicornis infestation. These results demonstrated that the rP27/30 protein might be a useful vaccine candidate antigen for biological control of ticks. PMID- 15785124 TI - Risk factors for retained placenta and the effect of retained placenta on the occurrence of postpartum diseases and subsequent reproductive performance in dairy cows. AB - The objectives of this study were to determine the risk factors for retained placenta by evaluating several reproductive factors in individual cows, and to determine the effects of retained placenta on the occurrence of postpartum diseases and subsequent reproductive performance in dairy herds. The health status, cow parity, calving, and breeding dates were recorded from 805 calvings in nine dairy herds from October 2000 to March 2004. We used logistic regression to evaluate the effects of these factors on the incidence of retained placenta. A stepwise procedure, used to obtain the appropriate model with alpha = 0.05, revealed that abnormal partus and gestation length were important risk factors for retained placenta. The occurrence of endometritis and metabolic disorders was greater (p<0.01) in the group with retained placentas than in the control group. The mean intervals from calving to first service and conception were prolonged (p<0.05) by 7 and 18 days, respectively, in the group with retained placentas compared to the control group. These results suggest that abnormal partus and gestation length are strongly correlated with the development of retained placenta, which increases the occurrence of postpartum diseases and subsequently decreases reproductive performance in dairy herds. PMID- 15785125 TI - Transcranial Doppler ultrasound analysis of resistive index in rostral and caudal cerebral arteries in dogs. AB - Transcranial Doppler (TCD) was carried out to determine the resistive index (RI) values of normal canine cerebral arteries and its reproducibility and to evaluate the change of cerebral vascular resistance following diuretics administration. RI values of rostral cerebral artery (RCA) were compared between fontanelle window and temporal window. Normal ranges and reproducibility of the RI values were examined in the rostal cerebral artery (RCA) and caudal cerebral artery (CCA). And after administration of diuretics, TCD-derived RI values were measured at RCA and CCA. Cerebral vascular RI values of RCA and CCA were 0.55 +/- 0.05 and 0.55 +/- 0.03 in the normal dogs, respectively. There was no significant difference of RI between male and female; between fontanelle window and temporal window. Reproducibility of RI measurements between intraobserver and interobserver were relatively high. The RI of RCA and CCA were significantly increased 15 minutes after mannitol administration (p<0.01) and returned to baseline values by 30 minutes, but it did not significantly change after furosemide and saline administration. The results suggest that TCD is a useful test which can obtain reproducible results from any window and has the advantage of detecting subtle changes in cerebral vascular resistance. PMID- 15785126 TI - Mitral valve prolapse in Cavalier King Charles spaniel: a review and case study. AB - A 5 year-old spayed female Cavalier King Charles Spaniel was presented after a 3- to 5-day onset of severe respiratory distress. The dog also had a history of several episodes of syncope prior to presentation. A comprehensive diagnostic investigation revealed a midsystolic click sound on cardiac auscultation, signs of left sided cardiac enlargement in ECG and thoracic radiography, mitral valvular leaflet protrusion into left the atrium, decreased E-point-to septal separation (EPSS) and mitral regurgitated flow in echocardiography, all of which are characteristic signs of mitral valvular prolapse. After intensive care with diuretics [corrected] and a vasodilator with oxygen supplement, the condition of the dog was stabilized. The dog was then released and is being medicated with angiotensin converting enzyme (ACE) inhibitor with regular follow-up. PMID- 15785127 TI - Idiopathic intranuclear inclusion bodies in the renal tubular epithelia of Japanese quail (Coturnix coturnix japonica). AB - We report idiopathic intranuclear inclusion bodies in the renal tubular epithelia of two cases of among the 960 Japanese quail (Coturnix coturnix japonica) in the course of the acute oral toxicity and dietary toxicity test. Basophilic inclusion bodies were seen only in the nuclei of renal tubular epithelia. We could not classify our case into any adenovirus infection by clinical signs and lesions. The inclusion bodies were only identified as adenovirus-like particles based upon the electronmicroscopical features. PMID- 15785128 TI - Comparative efficacy of standard AGID, CCIE and competitive ELISA for detecting bluetongue virus antibodies in indigenous breeds of sheep and goats in Rajasthan, India. AB - The sero-prevalence of antibodies against blue tongue virus (BTV) in 408 local breeds of sheep in Rajasthan state in India was investigated using standard agar gel immunodiffusion (AGID) test. Maximum seropositivities of 11.3% (13/115), 10.7% (13/121), 7.1% (11/155) and 5.9% (1/17) were recorded in the Chokla, Magra, Nali and Pugal breeds, respectively. Out of 107 goat serum samples, 6 (5.6%) were AGID positive. The performance of the standard AGID, counter current immuno electrophoresis (CCIE) and the competitive enzyme-linked immunosorbent assay (cELISA) for the detection of serum antibody against BTV in indigenous breeds of sheep were compared. Out of 178 sheep serum samples tested, 17 (9.5%), 22 (12.3%) and 54 (30.3%) were positive for group-specific bluetongue antibodies by AGID, CCIE and cELISA, respectively. There was appreciable difference in the seroprevalence detected by AGID, CCIE and cELISA in clinically healthy and diseased sheep with regard to relative sensitivities and specificities of the tests with cELISA being highly sensitive and specific followed by CCIE and AGID test. It was concluded that these indigenous breeds of sheep may be a potential reservoir of BTV infection and cELISA should be routinely used for the detection of antibodies against BTV in these local breeds of sheep. PMID- 15785130 TI - Anatomical study on true hermaphroditism in an Indian pig (Sus scrofa domesticus). AB - A pig was confirmed to be a true hermaphrodite on the basis of gross and histomorphological studies of the genital organs. The genitalia was consisted of left ovary, oviduct, two coiled uterine horns, body of uterus along with right testis and an epididymis. Vagina and vulva were absent but male urethra with prostate gland was present. Grossly the size of all the genital organs appeared to be normal. Histomorphologically, testis and epididymis were underdeveloped as there was no clearcut spermatogenia and sertoli cells but Leydig cells were normal. The ovary presents normal histological features with some portion of testicular tissue. Degeneration of uterine epithelium was observed along with normal endometrial glands. PMID- 15785129 TI - Cystic endometrial hyperplasia and endometritis in a dog following prolonged treatment of medroxyprogesterone acetate. AB - An 8-year-old female Yorkshire Terrier was presented for investigation of reduced appetite, and occasional vomiting. She has been treated with medroxyprogesterone acetate (MPA) from past 3 year-old age for contraception. Abdominal sonography showed abnormal enlargement of uterus, and ovariohysterectomy was performed. Main gross findings of uterus were enlarged lesions in two areas of the left horn, which had thickened wall and yellowish sticky material in the lumen. Histopathologically, cystic endometrial hyperplasia (CEH) and endometritis were present in the thickened area. In this case, CEH and endometritis may be attributed to prolonged treatment of MPA. It was concluded that further study is needed to clarify the association of MPA treatment with age, its pathogenesis and abnormal uterine changes in dogs. PMID- 15785131 TI - Propecia: the goose that lays the golden eggs. PMID- 15785132 TI - Words common to dermatology and birds. PMID- 15785133 TI - Immunohistochemical profile of multicentric reticulohistiocytosis. AB - BACKGROUND: There is enough evidence to support the knowledge that multicentric reticulohistiocytosis (MR) is a histiocytic proliferative disorder; however, the type of histiocytes involved is not well established. OBJECTIVE AND METHODS: To study the nature of cells present in MR lesions by studying the immunohistochemical profile of three new cases and reviewing 23 cases reported in the literature. RESULTS: MR histiocytic cells are positive for vimentin, CD68, and CD45, negative for S-100 protein, CD34, and XIIIa factor, and weak reactors for thrombomodulin. Small activated histiocytes are MAC387 positive. Lymphocytes, mainly CD4+ cells, are found in MR infiltrates. CONCLUSIONS: The MR immunophenotypic pattern does not suggest a type I or type II dendrocyte or a Langerhans cell origin. On the other hand, it points to a different cell derived from the monocyte-macrophage line. CD4+ cells may be responsible for activating the proliferation of histiocytic cells. Small histiocytic MAC387+ cells are likely to become the MR multinucleated giant cells. PMID- 15785134 TI - DermLite II: an innovative portable instrument for dermoscopy without the need of immersion fluids. AB - The DermLite II is an improvement on a previous instrument allowing for dermoscopy to be carried out without the need of immersion oil. It consists of a magnifying lens encircled by light-emitting diodes that can be adjusted for polarization or can be customized by the manufacturer to produce colors of specific wavelengths for visualizing pigmentation and structures of various dermal depths. This new version of the DermLite makes it very convenient for the evaluation of not just pigmented lesions, but nonpigmented skin cancers, scalp disease, and vascular patterns. It can be attached to a camera to record images and has a retractable faceplate for use with immersion oil. PMID- 15785135 TI - Cutaneous signs and syndromes associated with internal malignancies. AB - Cutaneous changes may indicate the presence of or herald the incipient development of internal malignancies. The authors review 24 cutaneous signs, symptoms, diseases, and syndromes associated with internal malignancies. Early recognition of these cutaneous changes may allow for vigilance for, and prompt detection and management of, the underlying neoplasm. PMID- 15785136 TI - EVOCLIN (clindamycin phosphate foam, 1%). PMID- 15785138 TI - What's new in dermatologic lasers: fat--the new frontier? PMID- 15785137 TI - Cosmetic preservatives: friend or foe? PMID- 15785139 TI - Syphilis 2005: key issues. PMID- 15785140 TI - If corpses neither cough nor sneeze, then rescuers wear face masks for protection against, uh...? PMID- 15785141 TI - Community-acquired methicillin-resistant Staphylococcus aureus: skin infection presenting as an axillary abscess with cellulitis in a college athlete. AB - A healthy 19-year-old black man without any methicillin-resistant Staphylococcus aureus risk factors developed axillary boils after he began lifting weights at the university gym in Houston, TX. He presented with a large tender erythematous fluctuant abscess of his right axillae and a superiorly located smaller painful red indurated nodule; the surrounding cellulitis extended into the adjacent tissue (Figure). The abscess was incised, drained, and cultured. Empiric treatment with cephalexin 500 mg q.i.d. was given for 7 days. The culture grew methicillin-resistant S. aureus. Susceptibility testing of the S. aureus isolate was performed by Laboratory Corp. of America (Houston, TX); the Vitek system (Biomerieux, Hazelwood, MO) was used, and the specimen was incubated for 8 hours. Confirmation of methicillin resistance was performed using a methicillin resistant S. aureus plate and the specimen was incubated for 24 hours. In addition to resistance to methicillin, the bacterial isolate was also resistant to ciprofloxacin, erythromycin, and penicillin. The S. aureus strain had intermediate susceptibility to levofloxacin and was susceptible to clindamycin, gentamicin, rifampin, tetracycline, trimethoprim/sulfamethoxazole, and vancomycin. The infection persisted and the antibiotic was changed to double strength trimethoprim/sulfamethoxazole, taken twice daily for 15 days. In addition, topical care included lesional and intranasal application of mupirocin 2% ointment and daily cleaning of the area with 10% povidone-iodine liquid soap. The skin infection completely resolved without recurrence within 2 weeks. PMID- 15785142 TI - Botulinum toxin-induced resolution of axillary granular parakeratosis. AB - A 44-year-old woman presented with an 18-month history of a pruritic dermatitis in the axillae bilaterally, beginning in the springtime. There was no history of significant hyperhidrosis. The patient did not recall any change in deodorant, soap, detergent, or other topical products. Her family history was unremarkable for anyone with similar symptoms. Physical examination revealed hyperpigmented plaques on an erythematous base in both axillae. The inframammary and intertriginous folds were free of lesions. Histologic examination taken from the left axilla was notable for mild digitate papillomatosis, compact hyperparakeratosis with large nuclei, and retained keratohyaline granules within the stratum corneum (Figure), consistent with axillary granular parakeratosis. A periodic acid-Schiff stain was negative for hyphae. Patch testing utilizing the thin-layer rapid-use epicutaneous test (T.R.U.E. Test, Mekos Laboratories, Hillerod, Denmark) was unremarkable. The patient was initially treated with fluticasone propionate 0.05% cream without notable improvement. She then received Clostridium botulinum type A neurotoxin injections to the axillae (50 U/axilla) with complete resolution of the rash within a few days. At 6-months' follow-up, the patient remained asymptomatic. PMID- 15785144 TI - Introduction. Management of atrial fibrillation in elderly patients. PMID- 15785145 TI - Pharmacologic therapies for atrial fibrillation. AB - Pharmacologic therapy for atrial fibrillation may be used for acute cardioversion or ventricular rate control or for long-term therapy to maintain sinus rhythm or control ventricular rates in atrial fibrillation. Therapies must be tailored to elderly patients, with particular attention to structural heart disease, bradycardia, hypotension, and other comorbidities, including renal or hepatic insufficiency. Such considerations may dictate the use or avoidance of certain agents. Other important considerations for elderly patients include challenges associated with anticoagulation and maintaining therapeutic international normalized levels without risk of bleeding. When considering pharmacologic agents for elderly patients, it is also valuable to take into account socioeconomic issues, such as access to medications, adherence to complicated dosing schedules, and availability of appropriate clinical follow-up to assess therapeutic efficacy and adverse reactions to various agents. A carefully constructed therapeutic regimen can provide effective symptom control and atrial fibrillation management for elderly patients. PMID- 15785146 TI - The epidemiology of atrial fibrillation in elderly persons: the tip of the iceberg. AB - Atrial fibrillation is the most common clinically relevant arrhythmia and preferentially afflicts elderly persons, especially persons aged >/=75 years, with associated complications of ischemic stroke and other adverse outcomes. More than 2.2 million Americans currently have atrial fibrillation, and this number is expected to increase by at least 2.5-fold over the next 50 years. With the aging of the US and international populations, as well as the rising prevalence of clinical risk factors, the incidence and prevalence of atrial fibrillation are rising rapidly. While improved adherence to existing risk-based therapies can reduce thromboembolic complications in elderly persons, additional research is needed to determine novel approaches to prevent the initial development of atrial fibrillation in order to decrease the individual and public health burden of this condition. PMID- 15785147 TI - Nonpharmacologic treatment of atrial fibrillation in elderly persons. AB - In recent years, new nonpharmacologic possibilities have emerged for the treatment of atrial fibrillation. The roles of surgery, radiofrequency catheter ablation, pacing, and atrial defibrillation in the treatment of atrial fibrillation are discussed. This text focuses on the interaction between different treatment modalities and the pathophysiologic mechanisms of atrial fibrillation and on the available data about the effectiveness in elderly persons. PMID- 15785148 TI - Rate control vs. rhythm control in the management of atrial fibrillation in elderly persons. AB - Antiarrhythmic medications used to maintain sinus rhythm have long been the treatment of choice in atrial fibrillation. The results of five prospective randomized trials comparing the efficacy and safety of rhythm-control to rate control strategies are now available. Reflecting the epidemiology of atrial fibrillation in the real world, most subjects enrolled in these investigations were elderly persons at increased risk of stroke or death. All of these trials have had similar results; these studies have failed to demonstrate a clear advantage of one treatment strategy over the other. A prespecified subgroup analysis among 3091 elderly patients in the Atrial Fibrillation Follow-up Investigation of Rhythm Management (AFFIRM) study revealed that rhythm control was associated with a higher risk of death than rate control. This review examines developments leading to and the implications and limitations of these trials and discusses recently issued practice guidelines and the justification for ongoing efforts to develop nonpharmacologic approaches to rhythm management in atrial fibrillation. PMID- 15785149 TI - Anticoagulation for atrial fibrillation in the elderly. AB - Atrial fibrillation is a risk factor for stroke, particularly among elderly patients. Multiple trials have established that antithrombotic therapy decreases stroke risk. Aspirin is associated with a relative risk reduction of about 21% and adjusted-dose warfarin (international normalized ratio 2.0-3.0) is associated with a relative risk reduction of about 68%. Warfarin is more effective than aspirin but is used less often than indicated because of hemorrhagic risk and the inconvenience of coagulation monitoring. The oral direct thrombin ximelagatran has been investigated for stroke prevention in patients with atrial fibrillation in two large clinical trials. The results suggest efficacy in a fixed dose compared with well controlled warfarin. Although anticoagulation intensity was not monitored or regulated during treatment with ximelagatran, it was associated with less bleeding than warfarin. Other antithrombotic agents are under development as alternatives to warfarin, but sufficient data are not yet available to justify their clinical use in patients with atrial fibrillation. PMID- 15785150 TI - Quality of life in older patients with atrial fibrillation. AB - This review summarizes what is known about quality of life (QOL) in older patients with atrial fibrillation (AF). The studies reviewed in this paper represent an increasingly broad repertoire of therapies for the treatment of AF and suggest that QOL in older patients does improve with treatment. The most dramatic improvements in QOL are noted in patients who are highly symptomatic and have poorer QOL at baseline. The data from studies where ablation and pacing therapy is used for treatment in patients with refractory AF vividly demonstrate this statement. There is also evidence of improvement in QOL in those with less severe symptoms, though it is extremely challenging to measure improvements in older patients who are asymptomatic (e.g., silent AF) or mildly symptomatic. Recommendations about new knowledge needed to optimize outcomes, particularly QOL, in patients with AF are based on these findings and the gaps in existing knowledge. PMID- 15785151 TI - Management of atrial fibrillation in patients with congestive heart failure. AB - An increase in the prevalence of both atrial fibrillation and congestive heart failure is occurring in part because of the demographic shift toward an aging population. These two clinical entities often coexist in the same patient, resulting in the worsening of symptoms and prognosis, rendering their management even more challenging. A large clinical trial is currently examining whether a rhythm-control strategy is superior to a rate-control strategy in patients with congestive heart failure. New nonpharmacologic therapies such as pulmonary vein isolation are undergoing clinical investigation and may also alter the management of these patients. At present, therapy for atrial fibrillation in patients with congestive heart failure should aim first at controlling heart rate to provide symptomatic relief; then sinus rhythm should be restored if symptoms persist or if congestive heart failure worsens. For a more definitive approach in managing these patients, the authors await further clinical trial data. PMID- 15785152 TI - Ethical issues in the management of geriatric cardiac patients. AB - Ethical Issue: When referred to hospice care the treating cardiologist believed that his patient would die in a few days. Instead, the patient lived over 18 months. PMID- 15785153 TI - Clinical trials: advanced age, anticoagulation intensity, and risk for intracranial hemorrhage among patients taking warfarin for atrial fibrillation. PMID- 15785154 TI - Electrocardiology teacher analysis and review: T wave alternans. PMID- 15785155 TI - Is there more to life than blood pressure? PMID- 15785157 TI - Left ventricular mass reduction in type 1 diabetic patients with nephropathy. AB - Left ventricular hypertrophy regression was postulated more likely to occur in diabetic patients when renal function was preserved. Seventeen type 1 diabetic patients followed for 12 months while receiving protocol-driven glycemic and blood pressure control had baseline and 12-month echocardiography. Despite identical treatment resulting in similar blood pressures, patients with better renal function (below the group mean, serum creatinine < or =1.7 mg/dL) demonstrated reduction in left ventricular mass and septal thickness as well as increase in left ventricular fractional fiber shortening not observed in those with worse renal function (above the group mean, serum creatinine >1.7 mg/dL). This latter group also did not experience the improvement in glycemic control observed in those with better renal function. Regression of left ventricular mass and functional improvement can be accomplished with improved glycemic control. In the presence of renal dysfunction, however, efforts to control glycemia and cardiac work are suboptimal. Aggressive glycemic and blood pressure targets to reduce cardiovascular morbidity in this high-risk population should be studied. PMID- 15785156 TI - The effects of losartan compared to atenolol on stroke in patients with isolated systolic hypertension and left ventricular hypertrophy. The LIFE study. AB - The Losartan Intervention For Endpoint reduction in hypertension (LIFE) study reported that a losartan-based antihypertensive regimen reduced cardiovascular morbidity and mortality (composite of cardiovascular death, stroke, and myocardial infarction) more than therapy based on atenolol in patients with left ventricular hypertrophy and isolated systolic hypertension (ISH). Patients aged 55-80 years with blood pressures 160-200/<90 mm Hg were followed for a mean of 4.7 years. Blood pressure was similarly reduced in the losartan (n=660) and atenolol (n=666) ISH groups. There were 88 (6.6%) patients who experienced a stroke, 18 of which were fatal. Of patients experiencing strokes, 72.7% had an ischemic stroke. ISH patients in LIFE compared to the non-ISH group had a higher incidence of any stroke and embolic stroke, and similar incidences of fatal, atherosclerotic, and hemorrhagic/other strokes. The incidence of any stroke (40% risk reduction [RR], p=0.02), fatal stroke (70% RR, p=0.035), and atherothrombotic stroke (45% RR, p=0.022) was significantly lower in losartan treated compared to the atenolol-treated patients. The 36% RR for embolic strokes in the losartan group was not statistically significantly (p=0.33) different from the atenolol group. These data suggest that losartan-based treatment is more effective than an atenolol-based treatment for patients with ISH and a high risk for stroke. PMID- 15785158 TI - Dietary influences on blood pressure: the effect of the Mediterranean diet on the prevalence of hypertension. AB - Hypertension has long been recognized as a major risk factor for several common cardiovascular diseases. The World Health Organization reports that the number of people with hypertension worldwide is estimated at 600 million, while 3 million will die annually as a result of hypertension. There is evidence to support that several factors related to lifestyle habits may influence blood pressure levels. Among these, the beneficial effect of diet on human health, as well as on the control of hypertension, has been underlined in several studies. Moreover, adherence to a Mediterranean diet has been associated with reduced all-cause mortality and, especially, coronary heart disease. The effect of this traditional diet on blood pressure levels has not been well understood and appreciated. This review summarizes the current understanding of the dietary influences on blood pressure control and the findings of observational and clinical studies that have evaluated the effect of the Mediterranean dietary pattern on the prevalence of chronic essential hypertension. PMID- 15785159 TI - Pheochromocytoma, thyroid disease, and hyperparathyroidism. PMID- 15785160 TI - Should we be evaluating blood pressure dipping status in clinical practice? PMID- 15785163 TI - New considerations relating to class effect with angiotensin-converting enzyme inhibitors--the PEACE study. AB - Angiotensin-converting enzyme inhibitor therapy provides positive outcome benefits in a number of cardiac scenarios including congestive heart failure, postmyocardial infarction, as well as in the hypertensive patient at cardiac risk. This benefit exists both in normotensive and hypertensive individuals and is present in those with various grades of cardiovascular risk. This beneficial cardiovascular effect has now been observed with several angiotensin-converting enzyme inhibitors, suggesting a class effect. The Prevention of Events with Angiotensin-Converting Enzyme Inhibition trial studied the effect of adding the angiotensin-converting enzyme inhibitor trandolapril to a contemporary therapeutic regimen of patients with stable coronary artery disease and preserved left ventricular function. In this study, the addition of trandolapril did not confer any additional benefit in terms of reducing the incidence of cardiovascular death, myocardial infarction, or coronary revascularization. The neutral findings in this trial add a new wrinkle to the concept of class effect for cardiovascular protection with angiotensin-converting enzyme inhibitors in patients with coronary artery disease. PMID- 15785164 TI - Ways of experiencing the life situation among United States patients with an implantable cardioverter-defibrillator: a qualitative study. AB - The purpose of this paper is to describe how a selected group of United States patients with an implantable cardioverter-defibrillator perceived their life situation. A qualitative design based on the phenomenographic approach was chosen to describe the patients' conceptions of their life situation. Fourteen patients eight men and six women, aged 21-84-were strategically selected to obtain as broad a variation as possible. The descriptive categories to emerge from the analysis of the interviews were trust, adaptability, and empowerment. The category labeled trust describes how patients trusted in the organization around them. The category labeled adaptability describes how patients adapted to living with an implantable cardioverter-defibrillator device. The category entitled empowerment describes how patients considered that they received support from family and friends as well as from health care professionals. This study suggests the need for a holistic intervention program comprising family, work, and leisure, focusing on patients' future life situation. PMID- 15785165 TI - Capacity for direct attention in patients undergoing percutaneous coronary intervention: the effects of psychological distress. AB - Emphasis is placed on educating coronary heart disease patients about lifestyle choices to reduce the risk of further cardiovascular events, but little attention has been paid to the impact of attentional capacity on patients' learning. The purpose of this study is to identify and compare the capacity for direct attention and the effect of psychological distress during hospitalization and 6 weeks after percutaneous coronary intervention. Forty subjects, mean age of 63 years, 27 men and 13 women, completed components of the Wechsler Memory Test to measure direct attention, and the Profile of Mood States. There were significant differences in the Digit Span Test-backward, a measure of working memory (t= 2.79; p<0.009), and the Profile of Mood States total score, a measure of psychological distress (t=3.183; p<0.003), from hospitalization to 6 weeks post percutaneous coronary intervention. Data demonstrate psychological distress and reduced capacity for direct attention during hospitalization indicating that hospital environments are suboptimal for learning. PMID- 15785166 TI - Safety and efficacy of resistance training in patients with chronic heart failure: research-based evidence. AB - Although a rich body of research exists regarding the safety and efficacy of resistance training, health care providers continue to caution patients with heart failure not to engage in this type of exercise. Research studies utilizing resistance training demonstrate improvements in muscular strength and endurance, New York Heart Association functional class, and quality of life. Despite the hemodynamic changes which occur during resistance exercise, no negative outcomes have been reported. The purpose of this paper is to review the most current research regarding the use of resistance training with heart failure patients to provide assistance to clinicians and enable them to provide education and appropriate recommendations to their patients. PMID- 15785167 TI - Supraventricular arrhythmias: an electrophysiology primer. AB - Supraventricular arrhythmias are the most wide-spread group of arrhythmias and affect all age groups. Atrial fibrillation is the most common arrhythmic disorder and is even more prevalent among the elderly. Due to their prevalence, it is imperative for the clinician to be informed about these arrhythmias and treatment considerations. This paper presents a basic review of the incidence, pathophysiology, diagnosis, and treatment of supraventricular arrhythmias, along with gender differences, and discusses important implications for the health care provider. A summary of common electrocardiogram findings in supraventricular arrhythmias is presented along with a brief overview of pharmacologic agents. PMID- 15785168 TI - International perspectives on CV nursing: cardiovascular nursing in Finland. PMID- 15785169 TI - Vitamin E supplements: helpful or harmful? PMID- 15785170 TI - Can ST-segment changes be evaluated for acute myocardial injury in a patient with right bundle branch block? PMID- 15785172 TI - Development of pump-oxygenator systems and the origins of open-heart surgery: a personal memoir. AB - Intracardiac surgery requires the use of a pump-oxygenator to maintain life while the heart is taken out of its usual circuit. Open-heart surgery became practical with the introduction of perfusion systems in the early 1950s. Many factors merged at this time to initiate the beginnings of open-heart surgery. PMID- 15785173 TI - Specialty heart hospital care: a comparative study. AB - The objective of this study was to compare measures of MedCath heart hospital patient severity, quality, and Medicare-related expenditures for comparable services to a group of comparison heart hospitals. This analysis is relevant to stakeholders' concerns over the emergence of the specialty care hospital industry. The study incorporates Medicare data for seven MedCath hospitals as compared with 1192 hospitals that performed open-heart surgery in federal fiscal year 2001. The authors developed cardiac-specific patient severity measures based on All Patient Refined-Diagnostic Related Groups and all subsequent analyses were standardized for differences in case mix between MedCath and comparison group hospitals. Study results indicate that MedCath hospitals have higher cardiac case mix severity, fare better in indicators for quality of care, and provide care at less expense to Medicare than comparison group heart hospitals. These results imply that "cherry picking" arguments and quality-of-care concerns of the specialty care hospital industry critics do not seem applicable for MedCath hospitals. PMID- 15785174 TI - Robotic mitral valve surgery: a technologic and economic revolution for heart centers. AB - A renaissance in cardiac surgery is occurring. Cardiac operations are being performed through smaller incisions with enhanced technologic assistance. Specifically, minimally invasive mitral valve surgery has become standard for many surgeons. At our institution, we have developed a robotic mitral surgery program with the da Vinci telemanipulation system, which has recently gained Food and Drug Administration approval. This system allows the surgeon to perform complex mitral valve operations through small port sites rather than a traditional median sternotomy. Our techniques and initial results are reported. Despite procedural success, these devices are not inexpensive and hospitals will have to justify their purchase. The implementation of robotic surgery has forced us to compare costs and benefits compared with conventional cardiac surgery. Nevertheless, our desire for improved and less traumatic patient care will drive this new technology, which will serve as a good model for us to study over the next several years. PMID- 15785175 TI - The evolution of mitral valve surgery: 1902-2002. AB - The evolution of the surgical therapy of mitral valve disease emanates from original statements by British cardiologists in 1902 and anecdotal individual surgical cases in 1923 and 1925. Considerable amounts of experimental investigation during these years and after World War II in 1948 finally resulted in the widespread use of closed mitral commissurotomy, a successful therapy for noncalcified mitral stenosis. The history of mitral valve surgery then rapidly progressed with a variety of prosthetic and bioprosthetic valve devices, ultimately, to a considerable number of successful valve repair operations with prosthetic ring annuloplasty. The authors conclude with a discussion of the current status of minimally invasive mitral valve surgery, both by direct vision and robotic assistance. The entire evolution of thought and technique of mitral valve surgery is summarized in this paper from 1902-2002. PMID- 15785176 TI - Current status of mitral valve repair. AB - Mitral valve repair is the procedure of choice to treat mitral valve dysfunction. Advantages of mitral valve repair over mitral valve replacement include improved long-term survival, better preservation of left ventricular function, and greater freedom from endocarditis, thromboembolism, and anticoagulant-related hemorrhage. The feasibility and durability of mitral valve repair depend upon the etiology of mitral valve dysfunction. In degenerative and ischemic mitral valve diseases, valve repair is possible in 90% of cases and freedom from reoperation is high. In contrast, rheumatic valves are less amenable to repair and durability is limited. Most isolated valve operations can be performed in a minimally invasive fashion using a partial upper sternotomy. In patients with atrial fibrillation and mitral valve disease, the Maze procedure or pulmonary vein isolation is performed, eliminating atrial fibrillation and the need for long-term anticoagulation. PMID- 15785177 TI - Cardiac resynchronization therapy for the management of chronic heart failure. AB - While numerous pathophysiologic mechanisms may lead to the onset and progression of chronic systolic heart failure, a variety of electrophysiologic abnormalities seen in the setting of chronic left ventricular dysfunction may also contribute to the natural history of the disease. Atrial, atrial-ventricular, and inter- and intraventricular conduction disturbances may place the failing ventricle at a further mechanical disadvantage, thus contributing to the functional impairment and poor outcomes associated with chronic heart failure. In the early 1990s, attempts at treating patients with end-stage systolic heart failure using conventional pacing strategies met with equivocal results. However, this work did provide further insight into the electromechanical consequences of advanced heart failure and suggested that atrial-synchronized biventricular pacing, or cardiac resynchronization therapy, might provide better and more consistent symptomatic and hemodynamic improvement. Several landmark clinical trials have evaluated the safety and efficacy of cardiac resynchronization therapy in New York Heart Association (NYHA) class III and IV heart failure. These studies have consistently shown statistically significant improvements in quality of life, NYHA functional class ranking, exercise tolerance, and left ventricular reverse remodeling. Some studies have suggested reductions in morbidity and mortality. This latter observation has been confirmed by a recent large-scale outcomes study. Thus, cardiac resynchronization therapy should be routinely considered in eligible NYHA class III and IV heart failure patients with ventricular dyssynchrony. PMID- 15785178 TI - Stent grafts for the treatment of abdominal aortic aneurysms. AB - Stent grafting for treatment of abdominal aortic aneurysms (AAAs) has been a major advance in endovascular surgery. Initial success with the original endoluminal stent graft encouraged worldwide study of the technology. In the United States, the Food and Drug Administration (FDA) insisted on considerable experience with the devices before approval because of early problems with device rupture, stent fracture, fabric perforation, graft migration, and modular separation. Complications associated with the endovascular graft technology led many to recommend its use only in patients who were considered at "high risk" for the standard, open procedure. Further study and device improvements have led to results that indicate the procedure has the potential to reduce operating time and blood loss and shorten intensive care unit and hospital stays compared with open surgical intervention. At present, there are three FDA-approved devices available for use, and a fourth is expected in 2003. The ultimate decision by the individual practitioner or the institutional team regarding which patients should be treated with endovascular technology is still not entirely straightforward. Patient selection should be based on vascular anatomy, the availability of a suitable device, the patient's desire for a minimally invasive procedure, and a commitment to what is likely to be a lifetime of device surveillance. PMID- 15785179 TI - Endovascular therapies for vascular disease. AB - Endovascular techniques including angioplasty, stenting, and endoluminal stent grafts represent important therapeutic options for the treatment of vascular disease. Technologic advances have allowed for the treatment of aneurysmal disease as well as extra-cranial carotid disease that previously required surgical methods. The success of various endovascular therapies varies based on anatomic location and extent of disease. The clinical results in different arterial segments are increasingly recognized in the published literature. The aortoiliac arterial bed appears to respond most favorably, with less favorable results observed in the infra-inguinal and infrapopliteal locations. There is increasing evidence that stent-supported carotid angioplasty using cerebral protection will play an important future role in the treatment of carotid artery stenosis. Less invasive techniques to treat abdominal aortic aneurysms with endoluminal stent grafts have dramatically changed the available therapeutic options. Improved devices and delivery systems will likely increase the number of patients who can be successfully treated in this manner. The evolution of endovascular therapies will continue to change the way we treat vascular disease. PMID- 15785180 TI - Potential use of high-resolution two-dimensional transthoracic echocardiography for visualization of the left anterior descending coronary artery. AB - High-resolution, two-dimensional transthoracic echocardiography (HR-2DTTE) can detect the difference in the left anterior descending coronary artery (LAD) wall thickness between patients with coronary artery disease and patients with normal coronary arteries. However, HR-2DTTE measurements of the LAD wall thickness are larger than measurements obtained by intravascular ultrasound and histology. This difference may be due to the inclusion of adventitia by HR-2DTTE. We evaluated the contribution of adventitia to the wall thickness of the normal and atherosclerotic LAD by comparing HR-2DTTE with high-frequency epicardial echocardiography. The LAD wall thickness was significantly greater in patients with coronary artery disease by both HR-2DTTE and high-frequency epicardial echocardiography. Both an increase in the intima plus media thickness and an increase in the thickness of adventitia contributed to the increase in the LAD wall thickness in patients with coronary artery disease. Adventitia represents a significant portion of the LAD wall thickness imaged by HR-2DTTE and its thickness increases significantly with the development of atherosclerosis. PMID- 15785181 TI - Alcohol septal ablation for obstructive hypertrophic cardiomyopathy. AB - Treatment of obstructive hypertrophic cardiomyopathy remains challenging. Medical therapy, surgical therapy, and pacemaker therapy have been shown to be beneficial in some patients over the years. Alcohol septal ablation is a catheter-based intervention that causes a controlled infarct in the septum leading to a decrease in the left ventricular outflow tract obstruction. Along with the impressive reductions in left ventricular outflow tract gradient, there is subjective and objective improvement in symptoms. Development of complete heart block requiring a permanent pacemaker after alcohol septal ablation remains a frequent complication, but has decreased in some recent studies. Alcohol septal ablation has been shown to improve diastolic function, decrease mitral regurgitation, decrease left ventricular hypertrophy and mass, as well as cause changes at the cellular and molecular level that improve myocardial function. Comparison of alcohol septal ablation to surgical myectomy has shown favorable results. Long term follow-up of alcohol septal ablation has shown sustained results out to several years. PMID- 15785182 TI - The AbioCor totally implantable replacement heart. AB - The AbioCor artificial heart (Abiomed, Inc., Danvers, MA) represents the latest technologic advancement in the quest for a total heart replacement system. The AbioCor is an electric heart with fully implantable components. The Food and Drug Administration approved a clinical trial in January 2001. The clinical trial was designed as an initial feasibility study to determine the safety and efficacy of this first generation system. The study criteria include end-stage adult heart failure patients who are not transplant candidates. These patients have biventricular failure with a predicted 30-day life expectancy of less than 30%. On July 2, 2001 the first AbioCor device was implanted. Six other patients have undergone implantation to date. Four of the seven have been successful as defined by the study parameters of 60-day survival with improved quality of life. Two patients were discharged from the hospital. Outpatient activities were possible in four patients. There have been no device malfunctions and no device-related infections. The trial is active and enrollment is ongoing. PMID- 15785183 TI - The evolution of the Heart Hospital of Austin: why we did it and what happened. AB - The Heart Hospital of Austin, Texas opened in 1998 after more than 2 years of planning and political struggles. A committed group of cardiologists had a vision of a physician-driven and patient-focused model of cardiovascular care. To ensure the success of the project the physician group grew and placed full-time cardiologists in surrounding communities. The physician group also concentrated on new programs to improve the cardiovascular health of Central Texans. Volume, efficiency, and quality indices point to the Heart Hospital of Austin as a leader in cardiovascular care. PMID- 15785184 TI - Crunch time for clinical cardiology. PMID- 15785185 TI - Atrial tachyarrhythmia with well-concealed 2:1 block-inadequacy of lead III as a "rhythm strip". PMID- 15785186 TI - The politics of Medicare reform. PMID- 15785187 TI - The heart hospital: flawed foundation or sound future? PMID- 15785188 TI - Is a chimera the answer? PMID- 15785189 TI - The Vanderbilt answer to crises in academia. PMID- 15785190 TI - High sensitivity C-reactive protein in clinical practice. AB - As a growing number of patients with low low-density lipoprotein cholesterol levels are diagnosed with atherosclerosis, research has shifted toward markers of inflammation in an attempt to improve global cardiovascular risk prediction. These markers include cytokines, cell adhesion molecules, and acute phase reactants like high sensitivity C-reactive protein, an innate immune response protein. When measured with new high-sensitivity assays, levels of high sensitivity C-reactive protein have proven to predict future cardiovascular risk at all levels of low-density lipoprotein cholesterol, at all levels of the Framingham Risk Score, and at all levels of the metabolic syndrome. Among apparently healthy men and women, levels of high sensitivity C-reactive protein of <1, 1-3, and >3 mg/L distinguish between those at low, moderate, and high risk for future cardiovascular disease, respectively. In clinical practice, high sensitivity C-reactive protein should be used along with lipid evaluation as part of global risk assessment. Improved knowledge of cardiovascular risk should lead to improved compliance with both lifestyle and pharmacologic interventions designed to prevent future cardiovascular events. PMID- 15785191 TI - Status of femoral closure devices. AB - Both sealing and suturing closure devices have been shown to shorten hemostasis time, reduce the discomfort of manual or mechanical compression, and allow for earlier ambulation after cardiac catheterization and percutaneous coronary interventions without increasing vascular complications compared with conventional compression techniques. Several studies now report a reduction in vascular complications for percutaneous coronary intervention patients treated with closure devices compared with manual compression with pronounced benefit seen in patients receiving glycoprotein IIb/IIIa receptor inhibitors. Adoption of a simple predeployment femoral angiogram is now standard practice for use of a closure device. In an attempt to develop devices that are safer and more "user friendly," considerable modifications and improvements have been made in newer generation devices. Ultimately, the acceptance of femoral closure devices will depend on which device provides a simple approach with reliable hemostasis and a cost that can justify their incorporation into routine practice. PMID- 15785192 TI - Trends and treatment of heart failure developing after acute myocardial infarction. AB - The development of heart failure and/or left ventricular systolic dysfunction following acute myocardial infarction remains prevalent despite improvements in the diagnosis and management over the past decades. Although transient in some cases, the majority of these patients continue to carry a substantially higher mortality risk. New data on the benefits of eplerenone and carvedilol from large clinical trials have added to the complexity of polypharmacy in this patient subgroup, although aggressive medical strategies to prevent or reverse infarct related left ventricular remodeling remains the most effective method to save lives and prevent disabilities. PMID- 15785193 TI - A strategy for patient access to innovation. PMID- 15785194 TI - The rise of the heart hospital and The Fall of the House of Usher. PMID- 15785195 TI - Physician ownership in specialty hospitals raises the bar for health care and the ire of Senator Breaux. PMID- 15785196 TI - Competition or collaboration: developing a chimeric practice model in academia. PMID- 15785197 TI - Conflicts of interest, integrity, and public respect: challenges to the professional standing of modern cardiologists. PMID- 15785198 TI - The Mid America Heart Institute: part II. 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 described the background and infrastructure of the Heart Institute. In part II, cardiovascular research and benefits of physician management are addressed. PMID- 15785199 TI - Ventricular septal rupture as a complication of myocardial infarction. PMID- 15785200 TI - The importance of heart rate response in myocardial perfusion imaging. PMID- 15785201 TI - Blocked atrial bigeminy. PMID- 15785202 TI - Coronary artery fistulas: case report and review of literature. AB - Congenital coronary artery fistula is a very rare malformation that may involve any or all coronary artery branches and any cardiac chamber. Elective closure of coronary artery fistulas by surgery or percutaneous transcatheter techniques is generally accepted in the presence of symptoms, but controversies exist in the management of asymptomatic patients. We describe a case where a fistulous communication was present between the proximal left anterior descending coronary artery and the pulmonary artery and provide a review of the available literature concerning management of patients with coronary artery fistulas. PMID- 15785203 TI - Precise characterization of the epitope recognized by a monoclonal antibody against Escherichia coli RNA polymerase. AB - We have recently isolated a monoclonal antibody directed against Escherichia coli RNA polymerase that does not inhibit transcription. This antibody is a useful tool to immobilize this enzyme for transcription assays or protein-protein interaction studies. The epitope of this monoclonal antibody was precisely located by a combination of protein deletion and synthetic peptide scanning. The amino acids of the epitope were also determined. We conclude that this antibody binds an epitope shared by several bacterial species and therefore can be used to characterize or purify other related enzymes. PMID- 15785204 TI - Development, characterization, and epitope mapping of a panel of twenty-four monoclonal antibodies specific for human inducible nitric oxide synthase. AB - A panel of monoclonal antibodies (MAbs) to human inducible nitric oxide synthase (hiNOS) has been developed. By isotype analysis of the MAbs cloned from the 24 different positive hybridomas, 13 were determined to be mouse IgG1, two were mouse IgG2a, two were mouse IgG2b, and the seven others were mouse IgM antibodies: all contained kappa light chains. The anti-hiNOS MAbs were initially characterized by ELISA, RIA, Western blot, and immunocytochemistry, and then they were epitope mapped using synthetic peptides and a three-step mapping procedure. In the first step, each of the 24 MAbs was tested by indirect ELISA for binding to 96 overlapping 18-amino acid-long peptides that span the entire 1153-amino acid length of hiNOS. Eight IgG class anti-hiNOS MAbs were found to bind to one of five different peptides. In the second step, a series of amino terminal and carboxyl terminal truncated peptides were synthesized for each of the five peptides to which one or more of the MAbs bound. Each of the eight anti-hiNOS MAbs was found to bind to the truncated peptides with a unique specificity that identified the amino acid segment involved in binding. The third step in the epitope mapping process utilized three series of overlapping 5-, 6-, 7-, 8-, and 9-amino acid-long peptides for each of these segments and identified the exact amino acids of hiNOS involved in antibody binding. Anti-hiNOS MAbs 2A1-F8, 2D2 B2, 21C10-1D10, and 24B10-2C7 were found to be especially useful in different immunoassays. PMID- 15785205 TI - Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C terminal end of soluble angiotensin-converting enzyme. AB - Angiotensin-converting enzyme (ACE) is a membrane-anchored ectoprotein that is proteolytically cleaved, yielding an enzymatically active soluble ACE. Two mouse monoclonal antibodies, MAbs 1B3 and 5C8, were generated to the C-terminal part of human soluble ACE. MAb 1B3 recognized the catalytically active ACE, as revealed by ELISA and precipitation assays, whereas Western blotting and immunohistochemisty on paraffin- embedded sections using MAb 5C8 detected denatured ACE. MAb 1B3 showed extensive cross-reactivity, recognizing 15 species out of the 16 tested. The binding of this MAb to ACE was greatly affected by conformational changes induced by adsorption on plastic, formalin fixation, and underglycosylation. Furthermore, MAb 1B3 binding to the mutated ACE (Pro1199Leu substitution in the juxtamembrane region, leading to a fivefold increase in serum ACE level) was markedly decreased. MAb 5C8 detected all the known expression sites of full-size ACE using formalin-fixed and paraffin-embedded human tissues. The sequential epitope for MAb 5C8 is formed by the last 11 amino acid residues of soluble ACE (Pro1193-Arg1203), whereas the conformational epitope for 1B3 is formed by a motif within these 11 amino acid residues and, in addition, by at least one stretch that includes Ala837-His839 located distal to the sequential epitope. Our findings demonstrated that MAbs 1B3 and 5C8 are very useful for the study of ACE shedding, for identification of mutations in stalk regions, and for studying alternatively spliced variants of ACE. In addition, binding of MAb 1B3 is a sensitive determinant of the integrity of soluble ACE. PMID- 15785206 TI - Generation of a monoclonal antibody agonist to toll-like receptor 4. AB - Toll-Like Receptor 4 (TLR4) has recently been identified as the lipopolysaccharide (LPS) receptor. Upon stimulation of TLR4 with LPS, a Th1 type cytokine response is observed. This immune modulation is thought to have a potential benefit in anti-tumor therapy. We have generated a TLR4 antibody agonist (5D24.D4) that mimics the action of the natural ligand. 5D24.D4 and LPS stimulation of TLR4 leads to a similar response, including IL-8 secretion, activation of NF-kB-mediated transcription, and the induced expression of an overlapping set of target genes. 5D24.D4 may be beneficial as an adjuvant anti tumor therapy. PMID- 15785207 TI - Preparation and characterization of monoclonal antibodies against nucleostemin, a protein that controls cell proliferation in stem cells and cancer cells. AB - Nucleostemin (NS) is a novel protein that promotes the proliferation of stem cells and cancer cells. The aim of this study is to prepare specific monoclonal antibodies against nucleostemin as a tool in nucleostemin research. To generate the antibodies, the cDNA coding nucleostemin was cloned from human gastric cancer cell line NCI-N87 with reverse transcriptase-polymerase chain reaction (RT-PCR). The partial cDNA fragment was inserted into prokaryotic expression vector pGEX 4T1 to make glutathione S-transferase (GST) fusion protein in Escherichia coli, and the whole-length cDNA fragment was cloned into vector pcDNA3 for expression in mammalian cells. The spleen cells, from the BALB/c mouse immunized with the GST-nucleostemin fusion protein, were used to prepare the monoclonal antibody with hybridoma technique. After screening with enzyme-linked immunosorbent assay (ELISA) and subcloning, six specific monoclonal antibodies against nucleostemin were obtained. Moreover, they also worked well in Western blotting, immunoprecipitation, and immunohistochemistry. These antibodies will be helpful in elucidating biological functions and the molecular mechanism of nucleostemin. PMID- 15785208 TI - Characterization of a monoclonal antibody that antagonizes the function of human endostatin. AB - Endostatin, a 20-kDa proteolytic fragment of collagen XVIII, is a potent inhibitor of angiogenesis and tumor growth. The anti-angiogenic effects of endostatin include inhibition of endothelial cell migration and proliferation, and inhibition of the activity of MMP2. Structure-function analysis of endostatin that implies this contravention function buried in separate fragments of endostatin introduces new issues into the understanding of the structure-function relationship of endostatin. We developed and characterized a novel murine MAb, 4E7, to human endostatin, which antagonizes the function of endostatin. As we show here, MAb 4E7 blocks the anti-migration/adhesion effects of endostatin in vitro and the anti-angiogenesis effect of endostatin in vivo, but the inhibition effect of endostatin on endothelial cell proliferation is not affected by MAb4E7. These results suggest that the anti-migration and anti-proliferation functions of endostatin may have distinct structural foundations. PMID- 15785209 TI - A chimeric antibody to varicella-zoster virus glycoprotein e. AB - Varicella-Zoster virus (VZV) immune globulin (VZIG) derived from pooled human serum is currently used in immunotherapy of VZV-associated complications of chickenpox and shingles. We developed a mouse-human chimeric antibody against a VZV glycoprotein E (gE) epitope as a safer replacement for VZIG. Variable (V) heavy- and V kappa light-chain exons, derived from an anti-VZV gE antibody secreting mouse hybridoma cell line, were cloned into expression vectors containing an immunoglobulin promoter and enhancer, and human IgG1 or kappa constant (C) region genes. The expression vectors were cotransfected into mouse myeloma cell line (NSO), generating transformants that secreted chimeric human mouse IgGs. The chimeric and the parent mouse antibody were indistinguishable in their antigen binding specificity. VZV gE chimeric antibody may prove to be a prophylactic antibody that could provide significant advantages over VZIG in having defined specificity, lessened possibility of contamination with viral pathogens, and consistent availability. PMID- 15785211 TI - Monoclonal antibodies. PMID- 15785215 TI - Telethinking. Interview by Vicki P. Glaser. PMID- 15785210 TI - Monoclonal antibodies reactive with the BAF155 (SMARCC1) and BAF170 (SMARCC2) components of human SWI/SNF-related complexes. AB - Mammalian SWI/SNF-related chromatin remodeling complexes are required for transcription controls that underlie differentiation, development, and tumor suppression. The complexes each consist of an ATPase of the SWI2/SNF2 family and approximately seven stably associated non-catalytic subunits. In spite of the importance of these complexes to biological processes, monoclonal antibodies to the various subunits have not been readily available. Mammalian complexes can vary in subunit composition, but the BAF155 (SMARCC1) subunit and a closely related protein, BAF170 (SMARCC2), appear to be ubiquitous components. Here we report the development of antibodies raised against a BAF155-derived peptide. The antibodies were raised against a single peptide of 18 amino acids. However, hybridomas expressing antibodies of two different specificities were isolated. One, designated DXD7, is specific for BAF155. The other, designated DXD12, is reactive with both BAF155 and BAF170. The antibodies are reactive against both native and denatured proteins, and are suitable for immunoprecipitation and Western blots. The DXD7 antibody is suitable additionally for immunofluorescence assays. PMID- 15785216 TI - University of Virginia Health System, Office of Telemedicine. PMID- 15785217 TI - Early outcomes of a care coordination-enhanced telehome care program for elderly veterans with chronic heart failure. AB - Veterans with chronic heart failure (HF) are frequently elderly, have numerous comorbid chronic medical illnesses, frequent hospitalizations, and have high rates of cardiovascular events. Within the Veterans Health Administration (VHA), primary care providers are required to manage the majority of HF patients because access to cardiac specialty care within the VHA may be limited. We designed and implemented a care-coordinated, nurse-directed home telehealth management program for veterans with difficult-to-manage or new onset chronic systolic HF. An in home telehealth message device was provided to the patient at enrollment, and patients received daily HF-specific education via the nurse coordinator and/or the device throughout their continuum of care. We collected demographic characteristics, clinical characteristics, and outcome data at the time of enrollment and at nearly 6 months after enrollment. A total of 92 patients were enrolled, with complete data available on 73. The mean patient age was 67 years, the mean left ventricular ejection fraction (LVEF) was 23%, and nearly all patients (99%) were men. After enrollment, significant improvements were found in blood pressure (129/73 to 119/69 mm Hg, p < 0.05), weight (196 to 192 pounds, p < 0.01), and shortness of breath rating (0-10 scale, 4.0 to 2.7, p = 0.02). Average daily doses of fosinopril (24 to 35 mg/d, p < 0.01) and metoprolol (84 to 94 mg/d, p = 0.05) were also improved. The total number of inpatient hospital days were reduced while on the home telehealth program (from 630 for the previous year to 122 for the duration of the program) with only 31% of the hospitalizations related to HF while on the program. Our nurse-directed, care coordinated home telehealth management program was associated with improved early outcomes in a group of elderly male veterans with chronic HF. PMID- 15785218 TI - Improving asthma control through telemedicine: a study of short-message service. AB - Home peak expiratory flow (PEF) measurement is recommended by asthma guidelines. In a 16-week randomized controlled study on 16 subjects with asthma (24.6 6.5 years old, asthma duration small ze, Cyrillic 6 months), we examined Global System for Mobile Communications (GSM) mobile telephone short-message service (SMS) as a novel means of telemedicine in PEF monitoring. All subjects received asthma education, self-management plan, and standard treatment. All measured PEF three times daily and kept a symptom diary. In the study group, therapy was adjusted weekly by an asthma specialist according to PEF values received daily from the patients. There was no significant difference between the groups in absolute PEF, but PEF variability was significantly smaller in the study group (16.12 +/- 6.93% vs. 27.24 +/- 10.01%, p = 0.049). forced expiratory flow in 1 second (FEV1; % predicted) in the study group was slightly but significantly increased (81.25 +/- 17.31 vs. 77.63 +/- 14.80, p = 0.014) and in the control group, unchanged (78.25 +/- 21.09 vs. 78.88 +/- 22.02, p = 0.497). Mean FEV1 was similar in the two groups both before and after the study. Controls had significantly higher scores for cough (1.85 +/- 0.43 vs. 1.42 +/- 0.28, p < 0.05) and night symptoms (1.22 +/- 0.23 vs. 0.85 +/- 0.32, p < 0.05). There was no significant difference between the groups in daily consumption of inhaled medicine, forced vital capacity, or compliance. Per patient, per week, the additional cost of follow-up by SMS was Euros 1.67 (equivalent to approximately $1.30 per 1 Euro), and SMS transmission required 11.5 minutes. Although a study group of 40 patients is needed for the follow-up study to achieve the power of 80% within the 95% confidence interval, we conclude that SMS is a convenient, reliable, affordable, and secure means of telemedicine that may improve asthma control when added to a written action plan and standard follow-up. PMID- 15785219 TI - Changes in diagnosis, treatment, and clinical improvement among patients receiving telemedicine consultations. AB - The aim of this study was to determine whether outpatient telemedicine specialty consultations to primary care clinicians result in changes in a patient's diagnosis, treatment management, and clinical outcomes. Medical records of patients who received two or more clinical telemedicine consultations in dermatology, psychiatry, and endocrinology were evaluated in a nonconcurrent retrospective analysis. Three indicators were used to measure changes in the processes of care and clinical outcomes: change in diagnosis, change in treatment, and patient clinical improvement. A retrospective review of 223 individual telemedicine patient medical records was conducted. Specialty telemedicine consultations were found to result in changes in diagnoses in 48% of the cases, changes in treatment therapy in 81.6% of the cases, and clinical improvement in 60.1%. These results are consistent with previous literature that has assessed changes in processes of care and outcomes from face-to-face specialty consultations in outpatient clinics. Changes in diagnosis and treatment therapy were found to be associated with clinical improvement with odds ratios (ORs) of 2.66 (95% confidence interval [CI]: 1.47-4.83) and 11.22 (95% CI: 4.49 31.48), respectively. This study found that telemedicine consultations resulted in changes in diagnosis and treatment regimens and also are associated with clinical improvements. PMID- 15785220 TI - Modifying the Roter Interaction Analysis System to study provider-patient communication in telemedicine: promises, pitfalls, insights, and recommendations. AB - This paper suggests modifications to the Roter Interaction Analysis System (RIAS) -the most widely used measure for assessing provider-patient communication during conventional face-to-face consultations--for use in telemedicine. The RIAS, which describes and categorizes communication behaviors, is used to quantify communication events, which may then be correlated with patient, provider, and system attributes and health outcomes. Most of the changes suggested here add new coding subcategories to characterize technology-related utterances and to provide opportunities for global assessments of the overall technology environment within which provider-patient interactions took place. There are also general issues raised that interaction analysis researchers should consider when studying provider-patient communication in a telemedicine context. These relate to nonverbal behavior, multiple participants, missing information, and validity and reliability. In addition to comparing telemedicine to in person consultations, a modified RIAS could be used to compare televideo consultations to each other, across different specialties and technical specifications. A modified RIAS would accommodate not only differences in the current technology environment, but also changes in the way providers and patients communicate over time. The more we know about what interaction patterns lead to best outcomes, the more emphasis can be placed on developing training programs and other interventions to enhance patient provider interactions in telemedicine. PMID- 15785221 TI - Delivering health care in rural Cambodia via store-and-forward telemedicine: a pilot study. AB - Since 2001, a monthly telemedicine clinic has helped provide health care to residents in a remote region in Cambodia. Physicians at Massachusetts General Hospital and Brigham and Women's Hospital in Boston, Massachusetts, and Sihanouk Hospital of HOPE in Phnom Penh, Cambodia, provide consultations via e-mail to a mobile nurse in the district of Rovieng, Cambodia. We describe the operations of the monthly clinic and report the results of a retrospective case review of the first 28 months of consultations. We also report the results of a satisfaction and willingness to pay survey. A total of 264 visits were made during the 28 monthly sessions. Mean duration of chief complaint at initial visit declined from 37 months to 8 months during the first and last 6 months of the study period, respectively. Thirty-six percent (n = 76) of new patients complained of abdominal pain. Nine percent (n = 20) of new patients were given an empiric diagnosis of goiter. The percent of patients requiring referral to a hospital outside of the village decreased over time. All patients surveyed were either "very satisfied" or "satisfied" with their care, and most patients were willing to pay for a visit, with a median amount of USD 0.63. We conclude that store-and-forward e mail consultative support for mobile nonphysician health care workers is a feasible model for delivering care in the developing world. Further research is needed to demonstrate improvement in health status, cost effectiveness, and sustainability. PMID- 15785222 TI - The current state of telemonitoring: a comment on the literature. AB - Telemonitoring, is defined as the use of information technology to monitor patients at a distance. This literature review suggests that the most promising applications for telemonitoring is for chronic illnesses such as cardiopulmonary disease, asthma, and heart failure in the home. Fetal heart rate monitoring and infant cardiopulmonary functions have also been monitored at a distance, as well as coagulation, or the level of activity of elderly people, assessed by the intelligent home monitoring devices. Hospitals, clinics, and prisons all have used telemonitoring, as have ambulances equipped with systems connected to the receiving hospital. Telemonitoring allows reduction of chronic disease complications thanks to a better follow-up; provides health care services without using hospital beds; and reduces patient travel, time off from work, and overall costs. Several systems have proven to be cost effective. Telemonitoring is also a way of responding to the new needs of home care in an ageing population. Real time monitoring of patients in ambulances reduces the time to initiate treatment and allows the emergency crew to be better prepared. The obstacles to telemonitoring development include the initial costs of systems, physician licensing, and reimbursement. In the future, virtual reality, immersive environments, haptic feedback and nanotechnology promise new ways in improving the capabilities of telemonitoring. PMID- 15785223 TI - Human and organizational factors affecting telemedicine utilization within U.S. military forces in Europe. AB - During the past 10 years, the United States Army installed telemedicine capabilities in more than 30 Army medical treatment facilities within the European Theater, including the former Yugoslavia. In spite of favorable reviews of utilization from the early days of the installations, there has been only limited use for patient care. We conducted a field study of the program implementation within a group of military clinics and hospitals using site visits and interviews. Problems leading to underutilization were identified primarily as organizational factors rather than classic "human factors" or "man-machine interface" problems. The majority of those interviewed felt that: they had not been given adequate information on the role of telemedicine systems in provision of health care; operational and support policies had not been developed adequately; cost-benefit of use was not clearly demonstrated; and that many organizational impediments existed. Additionally, the lack of strong clinician proponents was repeatedly cited. Although limited in scope, this study provides insight into the attitudes of clinicians given the opportunity to use various telemedicine modalities, and identifies organizational problems that could be rectified by a defined mission involving the use of telemedicine and a structure encouraging its acceptance in routine clinical practice. As a result of this study, recommendations were made for changes in implementation practices, which may have civil as well as military implications. PMID- 15785224 TI - Teleophthalmology-assisted corneal foreign body removal in a rural hospital. AB - This paper describes two cases of using telementoring from the Prince of Wales Hospital (POWH) Ophthalmology Department, Sydney, to Bourke Base hospital, a rural hospital in far west New South Wales, Australia. Eye trauma occurs frequently in a rural setting, and it can be difficult to assess the extent of the injury without a specialist ophthalmic service. One of the aims of the teleophthalmology is to facilitate the transfer of ophthalmic skills to local health providers such as general practitioners. We demonstrate that the removal of corneal foreign bodies and rust rings can be effectively supervised via teleophthalmology. PMID- 15785225 TI - Use of videoconferencing for depression research: enrollment, retention, and patient satisfaction. AB - The goal of this study was to describe the effects of using videoconferencing on participant enrollment, research measure administration and responses, study retention, and satisfaction. We recruited 400 patients from the Portland Veterans Affairs Primary Care Clinics for a randomized clinical trial of a care management intervention for depression. Patients recruited from distant clinic sites had the option of traveling to Portland, Oregon, for initial interviews or being interviewed using videoconferencing. Interviews included obtaining informed consent and administration of research measures. Remote participants were subsequently asked to complete a 12-item mail survey regarding the interview. There were no significant problems with the process of interviewing and obtaining informed consent by videoconferencing, as reported by patients and clinic staff. Twenty of the 31 participants interviewed by videoconferencing returned the satisfaction questionnaire. Participants indicated a high degree of satisfaction with these interviews, and expressed willingness to recommend videoconferencing to others. No differences were observed between the Patient Health Questionnaire depression scores of videoconferencing and in-person participants, and there was no significant difference in the 6-month rate of loss to follow-up in the randomized trial. Videoconferencing allows patients in rural and remote locations to participate in psychiatric research and expands sources of recruitment for research projects. PMID- 15785226 TI - An empirical chart analysis of the suitability of telemedicine for hospice visits. AB - Telehospice (TH) utilizes telemedicine technology to provide care at the end of life. A bistate project was launched in 1998 to study the use of home-based telemedicine for routine hospice care. Videophones were deployed for telenursing visits and evaluations by social workers. In order to determine what proportion of home hospice visits could be performed using currently available telemedicine technology, we reviewed clinical records retrospectively for hospice nurse home visits. Clinical notes documenting home nursing visits were obtained from two large hospices, one based in Kansas, and the other in Michigan. Records were randomly selected for patients who received hospice nurse visits during the month of January 2000. The charts were reviewed for patient demographic information, patient assessments, teaching activities, and interventions. Five hundred ninety seven nursing notes were analyzed using an 85-item coding instrument. After careful review of the records, the coders also made a subjective observation regarding the suitability of each visit for telemedicine. As part of this analysis, we found that 64.5% of home hospice nursing visits could be performed by telemedicine. Using TH, it is possible to reduce the number of in-person visits, thereby significantly decreasing the cost of providing hospice care. PMID- 15785227 TI - Telepsychiatry with child welfare families referred to a family service agency. AB - A telepsychology and telepsychiatry service, using ISDN interactive video H.320, providing psychological consultations for the Family Resource Center in Farmington, New Mexico, was evaluated. During the first year of service, 56 individuals participated including University of New Mexico staff, Family Resource Center staff and clients. Consultations involved children from families referred by the Children, Youth and Family Department (CYFD) in New Mexico. Telemedicine was used in some child abuse cases. This study investigated whether the quality and acceptance of telemedicine consultations were comparable to face to-face interactions in a group referred by a state child welfare agency. Participants received formal written consultation reports after the video conferencing sessions, which included a number of recommendations. Participants were asked to complete an anonymous questionnaire to rate various aspects of videoconferencing. The participants rated the university staff as involved, enjoyed the Family Resource Center staff's presence, felt the procedure was useful for evaluation purposes, found the format allowed for discussions of problems, and felt that the format was useful when compared to face-to-face consultations. The participants also said they had followed up on many of the recommendations. Videoconferencing appears to be a viable approach for providing consultation for families referred by a state child welfare agency. Several participants rated the session as both educational and consultative compared to simply therapeutic. PMID- 15785228 TI - The clinical interview and the doctor-patient relationship in telemedicine. AB - The doctor-patient relationship is one of the most complex interpersonal relationships, in that it involves individuals who are not on the same level, it has not been sought by both individuals, it is emotionally loaded and it requires close mutual co-operation towards a shared goal. A well-managed clinical interview lays the grounds for a good doctor-patient relationship which in turn leads to further advantages. In a classical ambulatory setting the doctor-patient relationship should take into account a number of context variables: physical environment, doctor's clothing and instrumental vs. affective behavior. The classical medical consultation takes place with the doctor and patient facing each other in the same room. Human contact, interaction and communication condition the course and treatment of the illness. This classical setting has been radically changed with the advent of videoconferencing. Video-communication reduces personal contact with the general doctor or specialist. Medical philosophy and the social sciences have often discussed the risk of alienating or "objectifying" the patient. The setting of a videoconferencing interview is completely different as interaction is not direct but occurs via a technological system that enables an unusual form of visual and auditory interaction that may be more or less realistic depending on the technology employed. The growing use of telemedicine and videoconferencing has led to new reflections on the doctor patient relationship in non-conventional settings and warrants in-depth studies on the dynamics of this form of consultation so as to increase the patients' level of satisfaction in telemedicine. The next step will be to verify experimentally and with adequate tools what the demands of patients are in a videoconferencing setting. PMID- 15785229 TI - Frontal and temporal morphometric findings on MRI in children after moderate to severe traumatic brain injury. AB - In vivo MRI volumetric analysis enables investigators to evaluate the extent of tissue loss following traumatic brain injury (TBI). However, volumetric studies of pediatric TBI are sparse, and there have been no volumetric studies to date in children examining specific subregions of the prefrontal and temporal lobes. In this study, MRI volumetry was used to evaluate brain volume differences in the whole brain, and prefrontal, temporal, and posterior regions of children following moderate to severe TBI as compared to uninjured children of similar age and demographic characteristics. The TBI group had significantly reduced whole brain, and prefrontal and temporal regional tissue volumes as well as increased cerebrospinal fluid (CSF). Confidence interval testing further revealed group differences on gray matter (GM) and white matter (WM) in the superior medial and ventromedial prefrontal regions, WM in the lateral frontal region, and GM, WM, and CSF in the temporal region. Whole brain volume and total brain GM were reduced, and total ventricular volume, total CSF volume, and ventricle-to-brain ratio (VBR) were increased in the TBI group. Additional analyses comparing volumetric data from typically developing children and subgroups of TBI patients with and without regional focal lesions suggested that GM loss in the frontal areas was primarily attributable to focal injury, while WM loss in the frontal and temporal lobes was related to both diffuse and focal injury. Finally, volumetric measures of preserved frontotemporal tissue were related to functional recovery as measured by the Glasgow Outcome Scale (adapted for children) with greater tissue preservation predicting better recovery. PMID- 15785230 TI - Neuroprotective anti-apoptosis effect of estrogens in traumatic brain injury. AB - Traumatic brain injury (TBI) is a leading cause of death and functional disability in western countries, affecting mostly young patients. Despite intense and sustained efforts deployed for the development of new therapeutic strategies, no clinical benefit has been shown by any of the investigated compounds. Increasing attention has been drawn during the past two decades to the neuroprotective effects of estrogens, although most of the available data relate to ischemic brain injury. The purpose of the present study was to investigate the potential neuroprotective value of estrogens in TBI as a therapeutic modality. For this purpose, a contusion was created in the parietal cortex by dynamic cortical deformation in two groups of 10 Sprague-Dawley male rats. Following the injury, treated animals received conjugated estrogens for 3 days, using a subcutaneously implanted osmotic pump. Animals were then sacrificed, and TUNEL, anti-active Caspase 3, bcl-2, and bax labeling were performed in paraffin embedded brain sections, allowing for comparative and quantitative analysis. In estrogen-treated animals, there was a marked and significant reduction of apoptosis in comparison with non-treated animals. The reduction in TUNEL and active Caspase 3 staining was similar and close to 50%. Optical analysis of histological slides prepared by bcl-2 labeling showed a significant increase in bcl-2 expression in estrogen-treated animals compared to non-treated animals. On the contrary, bax expression was not influenced by hormonal treatment, and no difference could be noticed between the two groups. These results support the potential therapeutic value of estrogens in TBI and further clarify their mode of action. PMID- 15785231 TI - VEGF and VEGF receptor expression after experimental brain contusion in rat. AB - Angiogenesis following traumatic brain injury (TBI) may be of importance not only for post-traumatic reparative processes but also for the development of secondary injuries. Vascular endothelial growth factor (VEGF) is a major regulator of endothelial cell proliferation, angiogenesis, and vascular permeability, though its possible involvement in secondary injuries after TBI is largely unknown. This study was undertaken to analyze the expression of VEGF and the VEGF receptors in experimental brain contusion in rat. Twenty-three adult female Sprague-Dawley rats were subjected to a focal cerebral contusion injury by use of a weight-drop model. Four additional rats underwent craniotomy only. The animals were sacrificed 6 h, or 1, 2, 4, 6, 8, or 16 days post-injury. Expression of VEGF and the VEGF receptors VEGFR1 (Flt-1) and VEGFR2 (Flk-1) were studied by in situ hybridization and immunohistochemistry. VEGF messenger (m)RNA and protein expression were detected in astrocytes, neutrophils, and macrophages in or adjacent to the injury from 1 day after injury, with a peak expression after 4-6 days. Flt-1 and Flk-1 mRNA and protein were detected in vessels adjacent to the lesion from 1 day after injury throughout day 6 after injury. It was also noted that Flt-1/Flk-1 and VEGF-positive vessels often were negative for SMI-71, a marker for vessels in areas with blood-brain barrier (BBB). In conclusion, we have demonstrated that TBI leads to an upregulation of VEGF, Flt-1, and Flk-1 mRNA and protein in and around the lesion. The data provide a foundation for future pharmacological intervention studies focusing on posttraumatic angiogenesis and possible injury repair effects of the VEGF system in TBI. PMID- 15785232 TI - The effects of gender on clinical and neurological outcomes after acute cervical spinal cord injury. AB - The potential clinical relevance of gender on clinical and neurological outcome after spinal cord injury (SCI) has received little attention. In order to address this issue, we examined all consecutive cases of acute traumatic cervical SCI admitted to our institution from 1998 to 2000. There were 38 males (ages 17-89 years, mean of 51.6) and 17 females (ages 18-84 years, mean of 63.2). Both groups were comparable regarding level (C1 to C7) and severity of SCI (ASIA A to D) at admission. Age differences between the groups approached significance (p = 0.057), and thus this factor was treated as a covariate in the analysis. Co morbidities were as frequent in men (86.8%) as in women (76.5%). The therapeutic approaches, length-of-stay in the acute care unit, mortality, and discharge disposition were similar in men and women. During hospitalization, 44.7% of men and 52.9% of women developed post-SCI secondary complications without any significant gender-related differences. Both groups showed a similar incidence of infections, cardiovascular complications, thromboembolism, and pressure sores. Univariate analysis revealed a trend for higher incidence of psychiatric complications (p = 0.054) and deep venous thrombosis (p = 0.092) in women, which was confirmed by multivariate analysis. Neurological outcome was not correlated with gender. A similar number of males and females (42.1%, 47.1%) showed evidence of neurological recovery as revealed by an improvement in ASIA scores. Moreover, 18.4% of males and 29.4% of females recovered to ASIA E status. Our data suggest a shift in the demographics of acute SCI with an increasing incidence in elderly women. Although neurological outcomes were not significantly related to gender, we observed a trend for higher rates of reactive depression and deep venous thrombosis in women. These issues may be of key clinical importance in developing improved management protocols for SCI so as to maximize functional recovery and quality-of-life. PMID- 15785233 TI - Upregulation of complement inhibitors in association with vulnerable cells following contusion-induced spinal cord injury. AB - We have previously described the activation of the classical, alternative, and terminal complement cascade pathways after acute contusion spinal cord injury using the New York University (NYU) weight-drop impactor. In the present study, we examined the induction of protein regulators of the complement cascade, factor H (FH), and clusterin, in the same experimental paradigm. The spinal cord of laminectomized adult rats was subjected to mild or severe injury using impactor weight-drop heights of 12.5 and 50 mm, respectively. The spinal cords of control and injured animals were evaluated at 1, 7, and 42 days after injury. Immunocytochemistry revealed a robust increase in the numbers and intensity of staining of FH, and clusterin-positive cells in the injured cord at all three time points, with the highest increases observed at 1 and 42 days after injury. FH and clusterin-positive cells were observed among neurons as well as oligodendrocytes. The increased expression was detected both rostrally and caudally from the injury site, in the latter case at distances up to 20 mm. The precise biological significance of injury-induced upregulation of these proteins remains to be determined. However, FH and clusterin are potent regulators of complement activity targeting upstream (FH) and downstream (clusterin) molecules of the pro-inflammatory cascade, which could be of vital importance in preventing a "runaway" inflammatory reaction in the injured spinal cord. PMID- 15785234 TI - Calpain inhibitors prevent neuronal cell death and ameliorate motor disturbances after compression-induced spinal cord injury in rats. AB - Traumatic spinal cord injury (SCI) results in widespread neuronal cell death. Recent studies have suggested that activated calpain mediates neuronal cell death in the central nervous system. We conducted a study to determine whether calpain mediates neuronal cell death in the motor neurons of the spinal cord after SCI, and whether postinjury administration of the calpain inhibitors N-acetyl- Leu-Leu Met-CHO (ALLM) and calpain inhibitor III (CI III) (MDL28170) reduces the motor disturbances in rats with a model of SCI. Adult male Wistar rats were subjected to SCI by application of a 20-g weight impactor probe to the spinal cord at T12 for 20 min. The rats were divided into three groups according to whether they were injected intravenously with 0.05-2.5 mg/kg ALLM, 10 mg/kg CI III, or 0.1% DMSO as a control every 24 h for 1 week after SCI. Calpain was activated in the spinal cord at 8 h, 24 h, and 5 days after SCI, and administration of ALLM inhibited its activation. ALLM, as compared to the DMSO vehicle alone, also significantly reduced the number of motor neurons in spinal-cord lesions that were positively labeled at 24 h after SCI with the terminal deoxynucleotidyl transferase-uridine nucleotide end-labeling (TUNEL) technique. Additionally, both the inclined plane test and footprint analysis showed markedly better motor activity after 4 weeks in rats injected with ALLM or CI III than in rats given vehicle only. These results suggest that activation of calpain plays a critical role in the neuronal cell death that follows SCI, and that calpain inhibitors may have benefit in treating the motor disturbances that follow SCI. PMID- 15785236 TI - Thyroid surgery: who should do it? How should it be done? PMID- 15785235 TI - Axonal pathology in subarachnoid and intracerebral hemorrhage. AB - Electrically active axons degenerate in the presence of nitric oxide (NO) in vitro. High CSF NO concentrations have been observed in patients with hemorrhagic brain injury such as subarachnoid hemorrhage (SAH) and intracerebral hemorrhage (ICH). This study investigated the evidence for axonal injury in SAH and ICH and related this to CSF NO levels. In this study, neurofilament phosphoforms (NfH(SMI34), NfH(SMI35), NfH(SMI38), NfH(SMI310)), surrogate markers for axonal injury, and NO metabolites (nitrate, nitrite = NOx) were measured by ELISA in cerebrospinal fluid (CSF) from patients with SAH and ICH and from a group of controls. Injury severity was classified using the Glasgow Coma Scale, and survival was used as the outcome measure. Compared to the control group, a higher proportion of patients with SAH and ICH had elevated NfH(SMI34) levels from day 0 to day 6 (p < 0.001), elevated NfH(SMI35) levels from day 1 to 6 (p < 0.001), and elevated NfH(SMI310) levels at day 0, 1, 4, and 6 (p < 0.001). The NOx levels were higher in the SAH and ICH patients than in the controls (p < 0.05) and distinguished the non-survivors from the survivors (p < 0.05). No direct correlation was found for NOx with any of the NfH phosphoforms. This study provides evidence for primary and secondary axonal injury in patients with SAH and ICH, with non-survivors also having higher NOx levels. CSF NfH phosphoforms might emerge as a putative surrogate marker for monitoring the development for secondary axonal degeneration in neurocritical care and guiding targeted neuroprotective strategies. PMID- 15785237 TI - An interview with Lewis E. Braverman M.D. PMID- 15785238 TI - Expression, hormonal regulation, and subcellular localization of CCAAT/enhancer binding protein-beta in rat and human thyrocytes. AB - The expression pattern of CCAAT/enhancer binding protein-beta (C/EBP-beta) was investigated in thyroid cells and tissues. Translation of C/EBP-beta mRNA results in the production of two isoforms, liver-enriched transcriptional activating protein (LAP) and liver-enriched transcriptional inhibitory protein (LIP), the latter lacking the transactivation domain. We found that LAP and LIP are expressed in the rat thyroid gland and in the FRTL-5 and PCCL3 rat thyroid cell lines. Thyrotropin (TSH), insulin, and serum withdrawal from cultures of thyroid cells induced downregulation of LAP and LIP expression. Subsequent activation of the cyclic adenosine monophosphate (cAMP) and insulin signaling pathways reinduced both isoforms. Vectors expressing rat LAP and LIP were constructed to study the effect of C/EBP-beta isoforms on the activity of the sodium iodide symporter (NIS) promoter in PCCL3 cells. The cAMP-stimulated activity of the NIS promoter was decreased by overexpression of LAP, whereas LIP had no significant effect. Expression of C/EBP-beta was studied by immunohistochemistry in normal human thyroid and papillary cancer tissues. C/EBP-beta immunostaining was always restricted to the nuclei of the normal thyrocytes. In contrast, C/EBP-beta was expressed mainly in the cytoplasm of thyroid papillary carcinoma cells. These data suggest that this factor may play important roles in the regulation of thyroidspecific genes and processes, and that its functions are altered in human thyroid carcinoma. PMID- 15785239 TI - Peroxiredoxin 5 expression in the human thyroid gland. AB - Peroxiredoxin 5 (PRDX5) is a newly discovered thioredoxin peroxidase able to reduce peroxides that is implicated in antioxidant protective mechanisms. We report here its expression in the human thyroid gland. Twenty-seven human thyroid specimens were examined by immunohistochemistry. They included six normal thyroid tissues, five multinodular goiters, nine hot nodules, two Hurthle cell adenomas, and five thyroids from patients with Graves' disease. In the control tissue, PRDX5 expression was heterogeneous, being stronger in cubical functionally active follicular cells than in flat quiescent thyrocytes. It was diffuse in the cytoplasm, occasionally localized in inclusions that most likely corresponded to mitochondria. This feature was particularly marked in the Hurthle cell adenoma case. In multinodular goiters, hot nodules, and Graves' thyroids, the cytosolic labeling was enhanced compared to the control tissue and a signal was also detected in few nuclei. To determine whether the level of expression was different between multinodular goiters and hyperthyroid Graves' thyroids, PRDX5 immunoblotting was performed in these two respective tissues. We observed that PRDX5 expression was higher in the thyroid gland of patients with Graves' disease compared to multinodular goiters. In conclusion, our data show that PRDX5 is expressed in the thyroid gland where it could act as antioxidant. The level of expression is directly correlated with the functional status of epithelial cells, being higher in multinodular goiters, and even more pronounced in hyperthyroid tissues, such as Graves' disease. PMID- 15785240 TI - Gene expression profiles reveal that DCN, DIO1, and DIO2 are underexpressed in benign and malignant thyroid tumors. AB - To investigate the molecular events involved in the pathogenesis and/or progression of thyroid tumors, we compared the gene expression profiles of three thyroid carcinoma cell lines, which represent major tumor subtypes of thyroid cancer and normal thyroid tissue. Using cDNA array methodology, we investigated the expression of 1807 open reading frame expressed sequence tags (ORESTES), selected from head and neck tumor libraries generated through the Brazilian Human Cancer Project-LICR/FAPESP. We found that 505 transcripts were differentially expressed in the thyroid carcinoma cell lines. Using a more stringent criterion, transcripts underexpressed or overexpressed more than fivefold in 1 of 3 or 3 of 3 carcinoma cell lines, a list of 55 ESTs were detected. Five candidate genes were further validated by quantitative polymerase chain reaction (qPCR) in an independent set of 52 thyroid tumors and 22 matched normal thyroid tissues. DCN was found underexpressed in a high percentage of the follicular thyroid adenomas, follicular thyroid carcinomas, and follicular variant of papillary thyroid carcinomas. DIO1 and DIO2 were underexpressed in nearly all papillary thyroid carcinomas. These genes not only could help to better define a tumor signature for thyroid tumors, but may, in part, also become useful as potential targets for thyroid tumor treatment. PMID- 15785241 TI - Troglitazone, the peroxisome proliferator-activated receptor-gamma agonist, induces antiproliferation and redifferentiation in human thyroid cancer cell lines. AB - Troglitazone is a potent agonist for the peroxisome proliferator-activated receptor-gamma (PPARgamma) that is a ligand-activated transcription factor regulating cell differentiation and growth. PPARgamma may play a role in thyroid carcinogenesis since PAX8-PPARgamma1 chromosomal translocations are commonly found in follicular thyroid cancers. We investigated the antiproliferative and redifferentiation effects of troglitazone in 6 human thyroid cancer cell lines: TPC-1 (papillary), FTC-133, FTC-236, FTC-238 (follicular), XTC-1 (Hurthle cell), and ARO82-1 (anaplastic) cell lines. PPARgamma was expressed variably in these cell lines. FTC-236 and FTC-238 had a rearranged chromosome at 3p25, possibly implicating the involvement of the PPARgamma encoding gene whereas the other cell lines did not. Troglitazone significantly inhibited cell growth by cell cycle arrest and apoptotic cell death. PPARgamma overexpression did not appear to be a prerequisite for a response to treatment with troglitazone. Troglitazone also downregulated surface expression of CD97, a novel dedifferentiation marker, in FTC-133 cells and upregulated sodium iodide symporter (NIS) mRNA in TPC-1 and FTC 133 cells. Our investigations document that human thyroid cancer cell lines commonly express PPARgamma, but chromosomal translocations involving PPARgamma are uncommon. Troglitazone, a PPARgamma agonist, induced antiproliferation and redifferentiation in thyroid cancer cell lines. PPARgamma agonists may therefore be effective therapeutic agents for the treatment of patients with thyroid cancer that fails to respond to traditional treatments. PMID- 15785242 TI - CT60 single nucleotide polymorphisms of the cytotoxic T-lymphocyte-associated antigen-4 gene region is associated with Graves' disease in an Italian population. AB - Graves' disease (GD) is an autoimmune and polygenic disorder. Several studies have shown that human leukocyte antigen (HLA) class II and the cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) gene are involved in the genetic susceptibility. We performed a case control study on 150 patients with GD and 301 controls, matched for age and gender, to verify the association of three polymorphisms located in CTLA-4 region (A49G, [AT](n)-3'UTR, and CT60) and of HLA DRB1 and DQB1 loci with the disease in an Italian population. The prevalence of patients with GD carrying the G allele of CT60 was significantly higher compared to control subjects (p = 0.02, odds ratio [OR] = 1.82). The allelic frequency of the G allele of CT60 was also significantly higher in patients with GD (p = 0.02). The G allele frequency of A49G in patients was significantly higher compared to control subjects (p = 0.04). The 280 allele phenotype frequency of (AT)(n)-3'UTR was also significantly higher in patients (p = 0.04). The G allele of A49G, the G allele of CT60, and the 280 allele of (AT)(n)-3'UTR microsatellite were significantly increased in patients with GD with thyroid-associated ophthalmopathy (TAO) compared to controls (p = 0.04, p = 0.03, and p = 0.02, respectively), however, we did not find any significant difference between TAO and non-TAO patients. We also found the HLA-DRB1*03 allele to be associated with GD; interestingly, the association of the CTLA-4 markers was independent from the HLA DRB1*03 status. These results highlight the role of the CTLA-4 locus, in addition to HLA, in the susceptibility to GD. Inside the CTLA-4 region, CT60 appears to be the most associated polymorphism to GD, however, further studies are needed to identify the etiologic variant. PMID- 15785243 TI - Immunoregulatory and susceptibility genes in thyroid and polyglandular autoimmunity. AB - The etiology of autoimmune thyroid diseases (AITD) is based on genetic and nongenetic factors. Genome-wide screening and linkage analyses have identified several chromosomal regions that are linked to AITD. These are HT-1 (on chromosome 13q33) and HT-2 (chromosome 12q22) for Hashimoto's thyroiditis (HT), and GD-1 (chromosome 14q31), GD-2 (chromosome 20q11.2), and GD-3 (chromosome Xq21) for Graves' disease (GD). Several genes have been proposed as susceptibility or immunoregulatory genes. Most promising genes are those of the major histocompatibility complex (MHC) complex (chromosome 6), the cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) gene (chromosome 2), the CD40 (chromosome 20), the thyroglobulin gene (chromosome 8), and the autoimmune regulator gene (chromosome 21). This review summarizes evidence for pathogenetic involvement of several of these genes in various forms of autoimmune thyropathies. Most genetic data refer to GD, whereas less data are available for HT and thyroid-associated ophthalmopathy. Scarce data refer to AITD within the autoimmune polyglandular syndromes I and II. The realization of family studies in large samples from different populations might provide further insight in the genetic contribution to AITD. Data are also needed on the interaction among susceptibility genes. Finally, additional functional studies are warranted to clarify the possible role of allelic variants in the underlying pathogenic mechanisms of AITD. PMID- 15785244 TI - Distinct diagnostic criteria for ultrasonographic examination of papillary thyroid carcinoma: a multicenter study. AB - Recent advances permitting high-resolution ultrasonography have made ultrasonographic examination of nodular thyroid diseases an accessible examination for routine practice. However, diagnostic criteria for ultrasonographic examination of thyroid nodules are not surely established. To identify the optimal strategy for well standardized differential diagnosis of papillary thyroid carcinoma and benign nodules, we evaluated the significance of individual ultrasonographic characteristics of thyroid nodules in a multicenter study. Ten characteristics in ultrasonograms from 53 patients scored by 17 investigators from 15 centers were analyzed by t tests and logistic regression analyses. Between benign and papillary thyroid cancer groups, all characteristics but not size or multiplicity of strong echoes, which suggest calcifications, were significant parameters. Logistic regression analyses showed that border character, shape, and internal echo level are highly significant parameters (p < 0.0005). A multiple logistic regression showed to be the most important predictors of pathologic diagnosis. The diagnostic criterion with border character and internal echo level yielded 93% sensitivity and 92% specificity. In conclusion, univariate and multivariate analyses identified border character, shape, internal echo level, but not strong echoes (calcifications), as important characteristics in differentiating papillary thyroid carcinoma from benign nodules. These results will contribute to standardization of accurate ultrasonographic diagnosis of papillary thyroid carcinoma. PMID- 15785245 TI - High prevalence of c-RET expression in papillary thyroid carcinomas from the Korean population. AB - BACKGROUND: Activation of the RET proto-oncogene, located on the long arms of chromosome 10, contributes to the development of thyroid cancers in two different ways. First, somatic rearrangements of RET with variable activation genes are frequently found in papillary thyroid carcinomas. Second, germ-line point mutations are responsible for the development of medullary thyroid carcinomas and multiple endocrine neoplasia type 2 (MEN 2). There are several conflicting reports on the influences of RET expression and RET/PTC rearrangements on the clinical outcome of thyroid cancers. Therefore, the wild-type RET gene expression and RET/PTC-1, RET/PTC-2, RET/PTC-3 rearrangements were examined in thyroid carcinomas and other thyroid diseases. MATERIALS AND METHODS: Thirty-six papillary thyroid carcinomas (PTCs), 8 follicular thyroid carcinomas (FTCs), 4 anaplastic thyroid carcinomas (ATC), 7 follicular adenomas (FAs), 23 hyperplasias, 6 normal thyroid tissues, and 39 normal portions from each tumor were included in this study. Reverse transcription-polymerase chain reaction (RT PCR) and immunohistochemical analyses were used to identify the RET gene and RET/PTC rearrangements. RESULTS: From the RT-PCR analysis, 68.9% of the PTCs, a single case of FTC, and 22.2% of the hyperplasias expressed the RET gene. No RET gene expression was observed in ATCs, FAs, or normal thyroid tissues. One RET/PTC 1 and one RET/PTC-2 rearrangement were detected in the PTCs. No RET/PTC-3 rearrangement was detected in any specimen. The immunohistochemical results revealed that 66.7% of PTCs, 28.6% of FAs, and 18.2% of hyperplastic thyroid tissue specimens showed high levels of RET protein expression. Neither the normal thyroid tissues nor the FTCs and ATC, showed high levels of RET protein expression. The two methods are agreed in PTC and hyperplastic nodules, but not in FA and FTC. CONCLUSION: PTCs among Koreans rarely showed RET/PTC rearrangements, but commonly showed increased RET gene expression. Compared to earlier reports indicating that the expression of the RET gene was limited to PTCs, the RET gene was also expressed in hyperplasias in this study. PMID- 15785246 TI - Clinical value of different responses of serum thyroglobulin to recombinant human thyrotropin in the follow-up of patients with differentiated thyroid carcinoma. AB - In the present study we examined the clinical value of a differential response of thyroglobulin (Tg) concentration after recombinant human thyrotropin (rhTSH) stimulation (rhTSH Tg testing) and its correlation with (131)I uptake and whole body scanning (rhTSH-WBS) in 104 patients who had previously undergone near-total thyroidectomy and (131)I ablation for differentiated thyroid carcinoma (DTC). rhTSH Tg testing was considered negative for rhTSH-Tg less than 0.9 ng/mL, low positive for rhTSH-Tg of 1-5 ng/mL and high positive for rhTSHTg greater than 5 ng/mL. rhTSH Tg testing was negative in 70 patients, 1 of whom had a lymph-node metastasis, but no (131)I uptake. Seven patients had low positive rhTSH Tg testing and no (131)I uptake, but 2 of these patients had cervical lymph node metastases. Twenty-seven patients had high positive rhTSH Tg testing and (131)I uptake was detected in lung, bone, or mediastinum in 11. Imaging techniques (computed tomography [CT], magnetic resonance imaging [MRI], fluorine-18 2-fluoro 2-deoxy-D-glucose-positron emission tomography [FDGPET]) documented metastatic disease in 22. In conclusion, our results suggest that any rise in rhTSH-Tg, even at low level, should raise the suspicion of persistent or recurrent DTC. Patients with rhTSH-Tg at high level should be carefully evaluated, because DTC persistence is highly probable. TSH-WBS provides little adjunctive information. PMID- 15785247 TI - Repeated fine-needle aspiration of the thyroid in benign nodular thyroid disease: critical evaluation of long-term follow-up. AB - Fine-needle aspiration (FNA) is the most effective diagnostic means in the management of nodular thyroid disease. Therefore, we decided to evaluate the diagnostic reproducibility of FNA and the usefulness of repeated FNA in benign nodular thyroid disease. We evaluated 799 satisfactory aspirations performed in 306 patients (298 females) aged 14-84 years, followed for 2-12 years. FNAs were performed on an annual basis. Of all patients, 299 (97.71%) maintained benignity: 230 (75.16%) repeated the same cytologic pattern, and 69 (22.54%) showed diagnostic variations within benign pathologies. In 3 patients (0.98%) the diagnosis changed to suspicious and in 4 (1.30%) to papillary carcinoma in the whole group. Of these 4 cases, 1 was diagnosed in the second (0.57%) and 3 in the third repeat FNA (2.25%). In 6 cases in which increased nodular size was observed, no change to malignancy was detected, while 3 of the carcinomas found showed no changes in size. IN CONCLUSION: (1) The variation in cytologic diagnoses within benign pathologies can be attributed to the coexistence of nonuniform pathologies. Aspirations performed in different sites of the nodule would contribute to obtain a more representative specimen. (2) In the absence of suspicious clinical changes and with at least three FNAs confirming the cytologic diagnosis of benignity, the follow-up of nodules by FNA could be discontinued and (3) In our experience, the diagnostic change from benign to malignant after three repeat FNAs appears to indicate that it would be advisable to perform three procedures. PMID- 15785248 TI - Reference intervals of serum thyroid function tests in a previously iodine deficient area. AB - AIM: We undertook the present study to establish reference data for serum thyroid function tests in a previously iodine-deficient area. METHODS: Data from 4298 individuals, 20-79 years of age were available for the present analysis. Thyroid function (thyrotropin [TSH], free triiodothyronine [FT(3)], and free thyroxine [FT(4)]) and serum autoantibodies to thyroperoxidase (anti-TPOAb) were evaluated from blood samples. Thyroid structure and size were measured by ultrasound. RESULTS: A reference population was selected comprising 1488 persons (825 men) by excluding subjects with known thyroid diseases, and with yet unknown thyroid disorders such as goitre, inhomogeneous thyroid pattern, nodules, hypoechogenicity and anti-TPOAb seropositivity. Reference intervals for serum TSH, FT(3), and FT(4) were 0.25-2.12 mIU/L, 3.8-7.0 pmol/L, and 8.3-18.9 pmol/L, respectively. Reference serum TSH levels were not comparable to the reference values that were recently established for the U.S. population and most reference values slightly differed from the reference values provided by the manufacturers. CONCLUSIONS: The reference ranges of thyroid function tests in this formerly iodine-deficient region are distinct from the reference ranges that were established in areas with iodine sufficiency. Creating a reference population in the present setting should include thyroid ultrasound in order to exclude yet undiagnosed thyroid disorders. PMID- 15785250 TI - An unusual presentation of hypothyroidism. AB - We describe a case of a 42-year-old male, with a 5-year history of recurrent gum bleeding, easy bruising, and chronic fatigue who presented for management of possible bleeding complications prior to a planned dental procedure. After extensive work-up, the patient was diagnosed with acquired von Willebrand Disease and underlying hypothyroidism with a thyrotropin (TSH) of 321 mIU/L (0.35-5.5) and total thyroxine (T(4)) of less than 1 microg/dL (4.5-12.5). He was started on levothyroxine and therapy and when compliant with treatment, the mucosal bleeding and symptoms of hypothyroidism were resolved. A subsequent TSH and total T(4) samples were drawn and found to be 6.3 mIU/L and 4.1 microg/dL, respectively. PMID- 15785249 TI - Necrotizing glomerulonephritis and pulmonary hemorrhage associated with carbimazole therapy. AB - Methimazole, carbimazole, and propylthiouracil (PTU) are the mainstays of antithyroid drug therapy. Adverse effects of these drugs have been documented in less than 15% of patients undergoing treatment for hyperthyroidism. Common problems include fever, skin rash, urticaria, arthralgias, and arthritis. Vasculitis associated with antineutrophil anticytoplasmic antibodies (ANCA) has been reported on several occasions following treatment with PTU. However, vasculitis rarely appears to be associated with carbimazole. We report the clinical history of a woman with a necrotizing glomerulonephritis and pulmonary hemorrhage associated with carbimazole therapy. PMID- 15785251 TI - Antithyroid drug-induced agranulocytosis: how has granulocyte colony-stimulating factor changed therapy? AB - This study examined whether granulocyte colony-stimulating factor (G-CSF) is beneficial for the treatment of antithyroid drug-induced agranulocytosis. From January 1975 to December 2001, 30,798 patients with Graves' disease were treated with antithyroid drugs at Noguchi Thyroid Clinic & Hospital Foundation. During this period, 109 patients (0.35%) were found to have agranulocytosis caused by antithyroid drugs. In the symptomatic group, the recovery time from agranulocytosis was significantly shorter after the introduction of G-CSF (5.5 +/ 3.5 days, n = 19) compared to the symptomatic group before its introduction (9.2 +/- 4.4 days, n = 37, p < 0.01). In the asymptomatic group, the recovery time from agranulocytosis was significantly shorter after the introduction of G-CSF (2.3 +/- 1.9 days, n = 15) compared to the asymptomatic group before the introduction of GCSF (5.4 +/- 4.3 days, n = 34, p < 0.05). However, G-CSF therapy was ineffective in severe cases with granulocyte count below 0.1 x 10(9)/L and symptoms. We recommend that G-CSF therapy should be applied only in asymptomatic patients and symptomatic patients with granulocyte count above 0.1 x 10(9)/L, and not for symptomatic patients with granulocyte count below 0.1 x 10(9)/L. In conclusion, G-CSF therapy shortens the period of recovery from antithyroid drug induced agranulocytosis and benefits patients, except those with symptoms and a granulocyte count below 0.1 x 10(9)/L. PMID- 15785252 TI - Cervical spine metastasis mimicking thyroid bed uptake in follicular thyroid carcinoma. PMID- 15785255 TI - Rehabilitation medicine: the European perspective. PMID- 15785256 TI - Balance, mobility, and falls among community-dwelling elderly persons: effects of a rehabilitation exercise program. AB - OBJECTIVE: To assess the short-term effect of an exercise-based rehabilitation intervention on balance, mobility, falls and injuries. DESIGN: This randomized, controlled trial with repeated measures was performed at an outpatient rehabilitation center. Elderly, ambulatory, community-dwelling volunteers underwent 6 wks of supervised stretching, balance, endurance, coordination, and strengthening exercises. Controls attended seminars. Data were recorded for time and quality performance on a functional obstacle course and for self-reported falls and injuries. RESULTS: From baseline through 6-mo follow-up, participants in the exercise group (n = 122) significantly outperformed those in the control group (n = 83). The exercise group's functional obstacle course quality improved 2.3% postintervention and 1.57% at follow-up compared with 0.3% for the control group for each time period (P = 0.001). Functional obstacle course completion time improved 7.69% at postintervention and 8.35% at follow-up for the exercise group compared with 4.0% and 3.4% for the control group. Of baseline fallers in the intervention group, 87% (compared with 34.5% for the controls) reported no falls in the subsequent 6 mos. Of those reporting injuries in the 6 mos preintervention, 89.7% in the intervention group (compared with 55.6% for controls) reported no injury at 6 mos postintervention. CONCLUSIONS: Our intervention can improve functional performance and protect against falls and fall-related injuries. PMID- 15785257 TI - Manual treatment effects to the upper cervical apophysial joints before, during, and after endotracheal anesthesia: a placebo-controlled comparison. AB - OBJECTIVES: In this preliminary, placebo-controlled clinical trial, two different manual treatments were compared, spinal manipulation and postisometric relaxation, for dysfunctional motion segments of the upper cervical spinal column. The influence of the muscular portion on the joint-play restriction of a motion segment can be ignored in anesthesia, and the manual evaluation of this joint-play restriction must be focused on nonmuscular structures. By retesting in anesthesia, it is possible to examine whether mobilization and manipulation affect exclusively the muscular structures or also affect the other parts of the motion segment. Conclusions can be drawn about the superiority of one or both treatments and about the structural basis of the restricted joint play and its palpation. DESIGN: A total of 26 inpatients at the surgical or orthopedic department of the University of Rostock were examined manually at four testing times: before and after manual treatment, in anesthesia, and within 24 hrs of completing anesthesia. They were randomized into three groups: postisometric relaxation (mobilization), spinal manipulation (thrust technique), and placebo. RESULTS: A highly significant effect for both treatments was found posttherapeutically (P < 0.01) but not for placebo. In anesthesia, the treatment effect of spinal manipulation was further significant (P < 0.01) when compared with placebo. For postisometric relaxation, however, it was not (P = 0.160). A significant difference between spinal manipulation and postisometric relaxation was not found in anesthesia (P = 0.137). The treatment effect postnarcotically was further significant when compared with placebo only for spinal manipulation (P = 0.011). CONCLUSIONS: Both treatments are superior to placebo. Postisometric relaxation seems to affect mainly the muscular parts of the treated segments and less so the other parts, such as the joint capsule or the segmental affiliated ligaments and fascia. Spinal manipulation seems to influence all other segmental parts more effectively, and the treatment effect persists longer. A joint-play restriction cannot be an exclusively muscular tension phenomenon. Segmental motion dysfunctions show a high variability in their spontaneous course. PMID- 15785258 TI - Effect of occupational keyboard typing on magnetic resonance imaging of the median nerve in subjects with and without symptoms of carpal tunnel syndrome. AB - OBJECTIVE: To examine the effects of occupational keyboard typing on median nerve shape and T2 relaxation and on forearm muscle T2 in professional typists with and without symptoms of carpal tunnel syndrome. DESIGN: Based on the Levine Carpal Tunnel Syndrome Symptom Severity scale (LCTSS), 12 female professional typist volunteers were divided into asymptomatic (LCTSS < 1.3, n = 5) and symptomatic (LCTSS > 1.3, n = 7) groups. Magnetic resonance images were acquired from wrist and forearms of all subjects before, immediately after, and 8 hrs after 3 hrs of typing. Forearm muscle T2 and median nerve T2 cross-sectional area and long/short axis ratio were evaluated by blinded observers. RESULTS: There was no difference between groups in any measured variable before typing. Median nerve T2 increased and long/short axis ratio decreased in asymptomatic subjects after typing, but there were no significant changes in symptomatic subjects. T2 increased in finger flexor muscles after typing, but there was no difference in the pattern of muscle T2 changes between groups. CONCLUSION: In magnetic resonance images of the median nerve at the carpal tunnel, swelling and T2 increases from baseline are a normal response to typing and may be less likely to occur in subjects with symptoms of carpal tunnel syndrome. PMID- 15785259 TI - Use of the AAEM guidelines in electrodiagnosis of ulnar neuropathy at the elbow. AB - OBJECTIVE: The goal of this study was to determine if practice patterns in Washington State were altered by the development and publication of the American Association of Electrodiagnostic Medicine practice parameter for electrodiagnosis of ulnar neuropathy at the elbow. DESIGN: Medical records from 350 workers' compensation patients who underwent surgery for ulnar neuropathy at the elbow in Washington State were reviewed for the consistency of the electrodiagnosticians' report and the practice parameter. Statistical analysis (chi2 or Fischer's exact test) compared prepublication and postpublication patterns of test use. RESULTS: Thirty-one percent of the 153 studies (n = 47) were not consistent with American Association of Electrodiagnostic Medicine guidelines before its publication vs. 34% of the 138 studies (n = 47) after publication. This was not a statistically significant change (P = 0.63). CONCLUSION: Evaluation of the usefulness and dissemination of the guidelines is warranted. The practice parameter may need to be simplified and educational efforts extended to improve provider use of new guidelines. PMID- 15785260 TI - Improvement of muscle strength in osteoarthritic knee patients after intraarticular knee injection of hyaluronan. AB - OBJECTIVE: The objective of this study was to investigate whether the knee concentric and eccentric muscle strengths can be improved in patients with bilateral knee osteoarthritis after intraarticular hyaluronan injections. DESIGN: A total of 25 patients with bilateral knee osteoarthritis and with a radiographic Ahlback grading scale of I or II participated in this study. One session of intraarticular knee injection of hyaluronan was given to both knees (weekly intraarticular knee hyaluronan injections for a total of 5 wks). Knee concentric and eccentric muscle strengths were recorded between 10 and 90 degrees of knee flexion. The recordings were done on two angular velocities, 80 and 240 degrees/sec. This study took place in a tertiary medical center with a gait laboratory and a KIN-COM isokinetic dynamometer. RESULTS: An increase in concentric and eccentric muscle strength adjusted for body weight (P < 0.01) was observed in both knees, ranging between 5.1% and 27.7%. CONCLUSION: The decreased knee muscle contraction strength (concentric and eccentric) can be improved in knee osteoarthritis patients with an Ahlback grading scale of I or II after five weekly intraarticular knee injections of hyaluronan. PMID- 15785261 TI - Hyaluronic acid for the treatment of knee osteoarthritis: long-term outcomes from a naturalistic primary care experience. AB - OBJECTIVE: Intraarticular hyaluronic acid is indicated for patients with osteoarthritis of the knee. However, clinical experience, especially efficacy and adverse events, with repeated injection series in the long term are limited. DESIGN: Patients were referred to a large primary care center for management of osteoarthritis of the knee. All were naive to intraarticular hyaluronic acid therapy and met our entry criteria, including resting visual analog scale pain of > 45 mm, radiographic confirmation of unilateral knee grade 1-3 osteoarthritis, and willingness to receive intraarticular therapy. Patients received a three intraarticular injection series with Suplasyn (10 mg/ml, 2-ml injection) over 3 wks. Patients were instructed to return for consideration of repeat injection series based on their perception of pain restricting daily activity and a resumption of severity similar to their initial presentation. This prospective naturalistic cohort was followed for 6.7 yrs. Patients completed baseline assessment of rest and walking visual analog scale pain (primary efficacy variable), completed a 5-point categorical global satisfaction score, and recorded adverse events and concomitant therapeutic modality use at each study visit. Patients returned for consideration of a repeat injection series based on their perception of symptom severity and were eligible if their resting visual analog scale pain was > 45 mm. The three-injection series and data collection were repeated, and again, patients were given similar instructions regarding consideration of a third injection series. RESULTS: Of 897 referral patients, 537 (mean age, 68 +/- 8 yrs; mean duration of symptoms, 7.4 +/- 4.1 yrs) met our criteria, and only 21 patients did not return for a second injection series. The mean time between first and second series was 27 +/- 7 wks. The change in walking visual analog scale pain was significantly improved from baseline after the first series (81.3%, P < 0.001) and second series (86.7%, P < 0.0001). Similarly, resting visual analog scale pain was significantly decreased from baseline after the first (P < 0.001) and second (P < 0.001) series, and patient satisfaction was significantly improved with each injection series (P < 0.03 and P < 0.01). Very few adverse events were recorded and were limited to local pain and swelling. Use of concomitant therapeutic modalities at presentation for a second injection series included: nonsteroidal anti-inflammatory drugs/cyclooxygenase-2 medications (37%), acetaminophen (31%), oral nutraceuticals (12%), and physical therapy and bracing (12%). CONCLUSIONS: Intraarticular hyaluronic acid injections were highly effective in improving resting and walking pain in patients with osteoarthritis of the knee on a first and a second treatment series. Duration of symptom control was about 6 mos, and the therapy was highly satisfactory to patients and was associated with very few local adverse events and limited use of concomitant therapeutic modalities. These data support the potential role of intraarticular hyaluronic acid as an effective long-term therapeutic option for patients with osteoarthritis of the knee. PMID- 15785263 TI - Implementation of formal learning objectives during a physical medicine and rehabilitation sports medicine rotation. AB - OBJECTIVE: To develop and implement formal learning objectives during a physical medicine and rehabilitation sports medicine rotation and characterize resident experiences with the objectives over a 16-mo period. DESIGN: Prospective, including learning objective development, implementation, and postrotation survey. RESULTS: A total of 69 learning objectives were developed by physical medicine and rehabilitation staff physician consensus, including 39 core objectives. Eighteen residents completed 4-wk sports medicine rotations from January 2003 through April 2004. Residents completed an average of 31 total objectives (45%; range, 3-52), of which 24 (62%; range, 3-35) were core. Residents completed the highest percentage of knee (60%), shoulder (57%), and ankle-foot (57%) objectives and reported that objectives related to these areas were most effective to facilitate learning. In general, residents reported that objective content was good and that the objectives delineated important concepts to learn during the rotation. Seventeen of 18 residents indicated that the objectives should be permanently implemented into the sports rotation and that similar objectives should be developed for other rotations. Based on our experience and the recommendations of residents, the average resident should be able to complete approximately 30 objectives during a typical 4-wk rotation. CONCLUSIONS: Successful implementation of specific, consensus-derived learning objectives is possible within the context of a busy clinical practice. Our initial physician staff and resident experience with the objectives suggests that this model may be useful as a supplementary educational tool in physical medicine and rehabilitation residency programs. PMID- 15785265 TI - Toward a taxonomy of assistive technology device outcomes. AB - The advancement of assistive technology device (ATD) outcomes research requires a substantive taxonomy for ATD outcomes. This article describes the assumptions and principles that should underlie such a taxonomy. It advocates for an approach to classifying outcomes that would promote consistency in how ATDs are distinguished, based on their intended effect on the user. The approach is designed to accommodate the widest variety of ATD applications, reflecting combinations of user population, ATD type, service, and context for use. In essence, the provisional taxonomy proposes that ATD outcomes can be effectively operationalized from three vantages--effectiveness, social significance, and subjective well-being. It emphasizes a distinction between the proximal effect of ATDs and the more distal outcomes associated with their use. Because it promotes consistency in the language used for categorizing outcomes, the taxonomic approach should facilitate the development of ATD-specific causal models. The utility of a taxonomy in ATD outcomes research is discussed. PMID- 15785266 TI - Instrumental evaluation of gait modifications before and during intrathecal baclofen therapy: a 2-year follow-up case study. AB - Intrathecal baclofen therapy has been used for several years, despite the fact that long-term gait modifications in ambulatory patients have not been thoroughly investigated. A 31-yr-old male patient affected by hereditary spastic paraparesis was evaluated clinically and by gait analysis. Evaluations were made before and at 6, 12, 16, and 24 mos after implantation. The patient showed a clear improvement in self-selected speed, step and stride length, knee and ankle kinematics, and ankle kinetics. Moreover, the response observed on self-selected speed is consistent with the intrathecal baclofen dose administered. To our knowledge, this is the first report of a long-term instrumental gait analysis assessment of a patient receiving intrathecal baclofen. Gait analysis could be a reliable and feasible assessment tool to evaluate ambulatory patients receiving intrathecal baclofen therapy over time and to help clinicians in determining exact dose requirements. PMID- 15785267 TI - Dysphonia associated with shoulder steroid injection. AB - Glucocorticosteroid injections into the shoulder are commonly used in the treatment of rotator cuff tendinitis. These injections rarely result in any serious complications and are generally considered a safe therapeutic intervention. Despite the extensive application of this treatment, there have been no reports of dysphonia occurring after corticosteroid injection into the subacromial space. We report the case of a 66-yr-old man with dysphonia after undergoing subacromial glucocorticosteroid injection for treatment of rotator cuff tendinitis. PMID- 15785268 TI - Neuroimaging and clinical manifestations of bilateral temporal encephalopathy secondary to murine typhus infection. PMID- 15785269 TI - Surgical intervention and capsular contracture after breast augmentation: a prospective study of risk factors. AB - Epidemiologic data on local complications after breast augmentation are scarce. In particular, few prospectively collected data are available on modern breast implants on this issue. Using data from the Danish Registry for Plastic Surgery of the Breast, the authors examined determinants of surgery-requiring complications and capsular contracture grades III to IV among 2277 women who underwent cosmetic breast implantation from June 1999 through April 2003. During an average follow-up period of 1.6 years after implantation, 4.3% of these women (3% of implants) required secondary surgery as a result of short-term complications. The most frequent clinical indications for surgery were displacement of the implant (38%), capsular contracture grades III to IV (16%), ptosis (13%), and hematoma (11%). Overall, the authors found that inframammary incision and subglandular placement were associated with decreased risks of developing complications requiring surgical intervention, whereas implants larger than 350 mL increased the risk of such complications (relative risk [RR], 2.3; 95% confidence interval [CI], 1.3-4.0). Thirty-nine Baker III to IV capsular contractures were identified, of which 22 were treated surgically within the study period. Submuscular placement of the implant decreased the risk of capsular contracture grades III to IV (RR, 0.3; 95% CI, 0.2-0.8), whereas surgical routes other than inframammary and drainage of implant cavity were associated with increased risk of capsular contracture. Current surgical practices and modern implants used for breast augmentation produce fewer short-term complications than procedures and devices of the past. This prospective study indicates that surgical procedures are more important predictors for local (short-term) complications than implant or patient characteristics. PMID- 15785270 TI - Subincisional muscular coverage of expander implants in immediate breast reconstruction with pectoralis flaps. AB - Immediate breast reconstruction with expander implants is a safe, simple procedure that allows for a rapid physical and emotional postmastectomy recovery. When complications occur, the patient may be left with a prolonged reconstructive course. Such complications may result from thin mastectomy flaps and resulting marginal skin flap necrosis and implant exposure. Muscle coverage of the implant under the skin incision prevents such marginal necrosis of skin flap from becoming a factor in implant loss. This paper demonstrates a simple method for providing subincisional muscle coverage of expander implants with pectoralis muscle flaps. In this technique, a pocket is developed under the pectoralis muscle. The sternal origin of the pectoralis is released from the midsternal position to its inferior origin. The pectoralis muscle is then rotated inferior laterally and sutured to the dermis of the underside of the inferior mastectomy skin flap, thereby providing subincisional muscle coverage of the expander implant. During a 5-year period, 42 patients between the ages of 36 and 61 underwent breast reconstruction utilizing this technique. In these patients, there were 4 instances of marginal necrosis. In each of these cases, the implants did not become exposed, and all patients completed the expansion process without significant delay and underwent subsequent implant exchange without incident. Five-year follow-up has shown good esthetic results in all patients. PMID- 15785271 TI - Anatomic and clinical study of rectus abdominis musculocutaneous flaps based on the superior epigastric system: ipsilateral pedicled TRAM flap as a safe alternative. AB - This anatomic and clinical study supports the use of the ipsilateral transverse rectus abdominis musculocutaneous (TRAM) flap over the contralateral TRAM flap for breast reconstruction. The costomarginal artery was constant in these dissections and supplied a vascular pedicle to the rectus abdominis muscle. Radiologic studies confirmed the communications between the costomarginal artery and the deep epigastric system, the same as between the costomarginal artery and the musculophrenic and intercostal arteries. The results of the clinical series confirmed the costomarginal artery as an alternative or adjuvant vascular pedicle of the deep superior epigastric artery in cases of potential lesion of the latter. The lack of pedicle tension because of the shorter distance to be transposed allowed a greater versatility in flap shaping and positively affected its aesthetic result. This fact made the authors change from the contralateral to the ipsilateral pedicled TRAM flap as the first option for autologous breast reconstruction because of vascular security. PMID- 15785272 TI - Use of abdominal quilting sutures for seroma prevention in TRAM flap reconstruction: a prospective, controlled trial. AB - This study is designed to evaluate the effect of abdominal quilting sutures on the incidence of abdominal seroma formation in patients undergoing pedicled transverse rectus abdominis musculocutaneous (TRAM) flap reconstruction. It is theorized that the use of such sutures during closure of abdominal flaps will collapse dead space, thus preventing abdominal seroma formation. A total of 71 consecutive patients undergoing pedicled TRAM flap breast reconstruction were randomly assigned to receive abdominal quilting sutures or to undergo a standard abdominal closure. Primary outcome measures included: daily drain output for the first 3 postoperative days, time to drain removal, and seroma formation. Drain output per day decreased with the use of abdominal quilting sutures; however, the time to drain removal was not significantly affected. Most importantly, there was no significant decrease in the incidence of seroma formation with the use of abdominal quilting sutures in this series. PMID- 15785273 TI - Paraumbilical perforator-based pedicled abdominal flap for extensive soft-tissue deficiencies of the forearm and hand. AB - A new paraumbilical-based pedicled abdominal flap was used in 11 patients with extensive soft-tissue defects of the forearm and hand. With a relatively narrow pedicle, large flaps up to 5- x 14-cm can be raised. Another advantage of this flap is the comfortable position of the hand and forearm for the patient. The main disadvantage is the conspicuous abdominal scars like the other pedicled abdominal flaps. PMID- 15785274 TI - JetPeel: a new technology for facial rejuvenation. AB - JetPeel (Tav-Tech Ltd., Yehud, Israel) is a new device for cosmetic resurfacing of the facial skin. It uses a new technology of a 2-phase stream that creates a jet composed of gas (oxygen) and microdroplets of fluid (saline) accelerated to supersonic velocities. This jet impacts the skin, causing gentle and accurate cosmetic peeling. Our preclinical and clinical experience with the JetPeel for rejuvenation of the face is presented. In a group of 50 patients, we found the JetPeel technology to be a safe and effective new tool for the usual indications for facial peeling. It was most useful and effective for perioral peel. JetPeel can be used in combination with other resurfacing modalities such as chemical peeling or laser. Further investigation is needed to explore other applications of this technology, such as using different combinations of gases and fluids and transdermal transfer of medication dissolved in the peeling jet. PMID- 15785275 TI - The effect of releasing tip-supporting structures in short-nose correction. AB - Correction of a short nose has been regarded as one of the most challenging and at times vexing procedures in rhinoplasty. One surgical option used to prolong nasal length is the freeing of the alar cartilages from adjacent structures by dividing the nasal tip supporting tissues. Five fibrous connections are known to be important in maintaining the nasal tip shape: fibrous tissues between the upper lateral and lower lateral cartilages; the lateral border of the lower lateral cartilages at the pyriform aperture; the interdormal ligament and anterior septal angle; the footplate of the medial crus and septal cartilage; and the dermocartilaginous ligament. This study was designed to determine which of the fibrous connections providing nasal tip support offer the most effect of lengthening when these structures are divided. We performed 10 open rhinoplasties on fresh cadavers, and we sequentially divided the previously mentioned tip supporting structures, except the dermocartilaginous ligament. The mucoperichondrium of the upper lateral and septal cartilages was also elevated, in accordance with the usual order of being released in a short-nose correction procedure. We measured the distance between the anterior septal angle and tip defining points by using calipers while the middle crura of the lower lateral cartilages were stretched with a skin hook. We found that the most effective length was gained by severing the lateral crus from the upper lateral cartilages, and moderate gain was noted from the release at the pyriform aperture and mucoperichondrium of the upper lateral cartilage. Release of other tip-defining structures was not statistically effective. PMID- 15785276 TI - A simple and quantitative method for three-dimensional measurement of normal smiles. AB - Smile reconstruction is one of the most challenging fields in plastic surgery. An understanding of the normal smile is essential in planning surgery for facial paralysis. The purpose of this study was to suggest a simple and quantitative method for the 3-dimensional analysis of smiles and to establish a basis for smile reconstruction in facial paralysis patients. The smiles of 50 volunteers (25 men and 25 women) were observed using a 6-mm digital video camera. Each subject had 10 points marked on the lips and nasolabial creases. Video editing programs were used to capture images of the resting and maximum smiles. The opacity of the image of maximum smile was adjusted and was merged with the resting image so that the migration of the points could be observed in 1 image. The migration distance (in millimeters) and the direction (in degrees) between each point were measured in frontal and both lateral views, and thus 3 dimensional vectors (x, y, z scalars) were obtained. The scalar values were converted to the values of a spherical coordinate system (distance, angle alpha and beta). Our work may offer a simple and quantitative method for measuring the smile and accurate guidance for reconstructive surgery. PMID- 15785277 TI - A report of 12 cases of basal cell carcinoma arising in lesions following laser therapy. AB - Since the latter half of the 1990s, laser treatment has become a common practice in Korea. A group of patients diagnosed as having basal cell carcinoma (BCC) also had a history of prior laser therapy performed on undiagnosed cutaneous lesions. Clinical and histologic evaluations of these cases were performed retrospectively. Among the 138 patients who were diagnosed with BCC from 1997 to 2001, 12 lesions (8.7%, 3 males and 9 females) developed following previous laser treatment to remove undiagnosed lesions. The mean age of the patients was 55.6 years, which was younger than that of the general Korean population (62.7 years), and their noses were the most common site of lesion development. The lesions had disappeared grossly after laser therapy for 6 patients, but for the other 6 patients, their lesions had persisted or grown. The interval between the time of laser therapy and diagnosis of BCC ranged from 1 month to 3 years. The average interval was 18 months. Histologically, aggressive forms were more common in these cases as compared with the usual patterns of BCCs. Although we do not think that laser therapy causes the development of BCC directly, laser therapy does not seem to be adequate for undiagnosed lesions. And our conclusion is that the cutaneous lesions should be biopsied and thoroughly diagnosed and managed prior to laser therapy. PMID- 15785279 TI - Safety of combining abdominoplasty and total abdominal hysterectomy: fifteen cases and review of the literature. AB - Combined surgeries eliminate the need for 2 anesthetics and 2 hospitalizations, the postoperative period of convalescence is no longer than for either procedure alone, and the total cost is reduced significantly. However, the safety of combining procedures is an unanswered question. We present 15 patients that underwent combined abdominoplasty with total abdominal hysterectomy from January 1992 to July 2002. A retrospective chart review was performed; surgical reports were analyzed, along with subsequent clinical follow-up notes. One patient had a wound infection that required hospitalization, drainage, and intravenous antibiotics. Four patients had minor complications. Patients' satisfaction with the esthetic and functional results of combined procedures has been impressive. Our experience confirms the basic safety of the combined surgery when done on appropriately selected patients. These results are discussed in respect to the current literature. PMID- 15785280 TI - Repair of chronic mallet finger deformity using Mitek micro arc bone anchor. AB - Surgical correction of chronic mallet finger caused by terminal tendon disruption was carried out in 22 patients. The distal stump of the tendon was fixed to the base of the distal phalanx with a Mitek micro arc bone anchor. In all patients the mallet finger deformity was corrected. There were 15 patients with excellent results, 5 with good results, and 2 with fair results. None of the patients had a poor result. No further treatment was needed. The Mitek micro arc bone anchor system is a reliable alternative for the treatment of chronic mallet finger deformity without proximal interphalangeal hyperextension. PMID- 15785281 TI - The value of radial collateral ligament reconstruction and abductor digiti minimi release in metacarpophalangeal joint arthroplasty. AB - Rheumatoid arthritis is a chronic, progressive disease characterized by destructive synovitis commonly involving the hand. Ulnar drift deformity has been reported in up to 30% of these patients. Metacarpophalangeal (MCP) joint arthroplasty with soft-tissue reconstruction was described to correct this deformity, but recurrence is a common problem. This is a retrospective study of 18 patients who underwent 96 primary index to little finger MCP joint arthroplasties in Montreal General Hospital. Patients were divided into 2 groups. In group I, 60 MCP joints in 15 hands had silastic arthroplasty with radial collateral ligament reconstruction and abductor digiti minimi release. In group II, 36 MCP joints in 9 hands had silastic arthroplasty without soft-tissue reconstruction. All patients had the same postoperative rehabilitation, with a follow-up mean time of 63 months. Postoperative clinical and radiologic comparison was performed. Group I was found to have more severe wrist disease. No significant difference was seen between the 2 groups for ulnar drift (D2, P < 0.79; D3, P < 0.11; D4, P < 0.49; D5, P < 0.31), active range of motion, power grip, incidence of MCP subluxation, or functional ability. MCP arthroplasty with radial collateral ligament reconstruction and abductor digiti minimi release may recreate a short-term balance of forces around the MCP joint but does not prevent late postoperative deformity. This is a preliminary study with a small sample size but supported previous reports on MCP arthroplasty with soft-tissue reconstruction. PMID- 15785282 TI - A comparison of combinations of titanium and resorbable plating systems for repair of isolated zygomatic fractures in the adult: a quantitative biomechanical study. AB - Multiple studies have sought to determine the postreduction stability of internal fixation in zygomaticomaxillary complex (ZMC) fractures. Three-point fixation with titanium miniplates is increasingly recommended to repair these injuries. Use of bioresorbable plates has been suggested to eliminate potential postoperative hardware complications. By quantitatively comparing different combinations of titanium and resorbable plating systems, this study attempts to demonstrate which combinations will provide stable fixation of the fractured ZMC. Osteotomies were performed on 40 zygomas in 20 fresh-frozen cadaver skulls, simulating noncomminuted ZMC fractures. The control group (group 0) consisted of titanium plates at the zygomaticofrontal (ZF) suture, infraorbital rim (IOR), and zygomaticomaxillary buttress (ZMB). Group 1 consisted of titanium plates at the ZF and IOR, and a resorbable plate at the ZMB. Group 2 used a titanium plate at the ZF, and resorbable plates at the IOR and ZMB. Group 3 consisted of resorbable plates at the ZF, IOR, and ZMB. A mechanical test system was used to apply loads in the vectorial direction of the masseter. Critical forces and patterns of hardware failure were recorded. Group 0 failed at a mean force of 589 +/- 146 N (60 kg). Group 1 failed at a mean force of 507 +/- 124 N (52 kg). No statistically significant differences between groups 0 and 1 were found. The mean force required for failure in groups 2 and 3 was lower. Differences in the force required for failure between groups 2 and 3 and the control group was significant (P <0.05). Failure patterns were analyzed. The ZF plate tended to stretch predominantly in groups 1, 2, and 3, whereas it tended to break in group 0 (P = 0.005). The IOR plate demonstrated predictable screw failure in groups 2 and 3 (P = 0.007). For group 0, the ZF was the site of the majority of critical failures. For groups 2 and 3, the IOR was almost invariably the site of critical failure (P = 0.004). At the ZMB, there was no significant association between failure modes and it was rarely the site of critical failure, regardless of the method of fixation. However, the strength of fixation was proportional to the number of titanium plates used. Overall, the method of fixation significantly affected the force required for mechanical failure of ZMC fractures (P <0.0001). The presence of teeth significantly increases the force required for implant failure in ZMC fracture fixation when combinations of plates are used (P = 0.038). All combinations of titanium and resorbable plates may be sufficient to overcome the displacing forces produced by the masseter and may be used for internal fixation of isolated ZMC fractures in the adult. PMID- 15785283 TI - Comparative efficacy of ropivacaine and bupivacaine infiltrative analgesia in otoplasty. AB - A prospective double-blind study was conducted to compare the anesthetic efficacy of ropivacaine and bupivacaine in a bilaterally symmetrical otoplasty model. Because ropivacaine has a significantly lower toxic potential than bupivacaine, it may be established as the anesthetic agent of choice for low-dose infiltration anesthesia in routine aesthetic facial operations. Each of the 24 adult patients undergoing bilateral otoplasty received infiltration with ropivacaine in 1 auricle and an equal volume of bupivacaine in the other. Patients were requested to assess their pain separately in each side at the times of infiltration, during cartilage scoring, and 2, 6, and 10 hours postoperatively on a visual analog scale. Intraoperative success rates were similar, and overall analgesia achieved at 2 hours, 6 hours, and 10 hours postoperatively was not found to be statistically different between ropivacaine and bupivacaine. The authors conclude that ropivacaine can be used as an effective alternative to bupivacaine in otoplasty. PMID- 15785284 TI - Referral patterns and severity distribution of burn care: implications for burn centers and surgical training. AB - The purpose of this investigation was to examine burn-patient referral patterns and severity of burn distribution, as well as to determine the impact these patterns may have on the education of surgeons in training. Data from the 1998 1999 National Inpatient Sample (NIS) and the Michigan Hospital Association (MHA) were analyzed based upon burn diagnostic-related groups (DRGs; 504-511) and their referral distribution was documented. Providers were segregated into high-volume hospitals (HVHs) treating >100 patients per year, moderate-volume hospitals treating 25 to 99 patients per year, and low-volume hospitals (LVHs) treating <25 patients per year. Surgical training programs were identified within the state of Michigan and examined for an educational affiliation with a burn center. Across the United States, 47.5% of burn patients receive care at HVHs. Patients with the highest severity (ie, DRGs 504 and 505) were usually (77%) treated in HVHs. Within the state of Michigan, 4 HVHs were identified, which represent 50.8% of the total burn admissions. At least 1 HVH received over 80% of its admissions from adjacent or distant counties and subsequently represented a higher proportion of higher-severity burn DRG admissions. Twenty-three percent of general surgical programs within the state of Michigan do not have a formal burn rotation or affiliation with a regional burn center for educational training. Several programs have affiliations with low-volume burn providers. The most severe burns are reaching high-volume centers, but many burns continue to remain within LVHs. A wide variation in patient distribution occurs throughout the United States. Matching the patient and resident distribution is essential for effective training of surgical residents. PMID- 15785285 TI - Effect of FK506 on peripheral nerve regeneration through long grafts in inbred swine. AB - Numerous small-animal studies have demonstrated that FK506 enhances nerve regeneration and accelerates functional recovery after nerve injury. However, no experimental study has corroborated these neuroregenerative effects in larger animals. This study investigated the effects of FK506 on nerve regeneration in inbred miniature swine. Eight animals received 8-cm ulnar nerve autografts and allografts. Treated animals received 0.1 to 0.4 mg/kg FK506 injections twice weekly to maintain immunosuppressive serum FK506 levels. At 24 weeks posttransplant, nerve grafts were harvested for histomorphometric analysis. Mixed lymphocyte cultures demonstrated alloreactivity in 1 treated animal and all untreated animals. In autografts, mean fiber count, nerve density, and percent neural tissue were doubled with FK506 therapy. In allografts, significant neuroregeneration was observed in animals treated with FK506, whereas untreated animals had no regeneration. Treatment with FK506 resulted in a trend toward enhanced axonal regeneration through nerve autografts and allografts in a large animal model with defined histocompatibility barriers. PMID- 15785286 TI - Bone flap prefabrication: an experimental study in rabbits. AB - The usual method to prefabricate a bone flap is to harvest a nonvascularized bone graft and to implant the artery and vein bundle between segments of bone graft. The basic problem of this method is sacrificing an artery for prefabrication. Another method for creating flap donor sites without using an artery is venous flap prefabrication. There are a few articles describing bone flap prefabrication, and these include implantation of both artery and vein as a vascular bundle. Also, there is no experimental study in the literature using a vein or an arterialized vein pedicle for bone flap prefabrication. As an experimental model for bone flap prefabrication, the rabbit ear vascular model was chosen. For the experiments 3 groups were formed. Each group contained 5 rabbits. In the first experimental group a vein was implanted between the halves of bone graft. In the second experimental group an arterialized vein was implanted between the halves of bone graft. To compare the viability of the bone graft of the 2 prefabrication groups, a bone graft was implanted into the subcutaneous pocket of the posterior auricular area in the third group. The authors examined 5 rabbits in each group by microangiography at the end of 6 weeks except for group 3. On microangiographic analysis, groups 1 and 2 showed patency of the vascular pedicle. There was no difference between these 2 groups from the point of view of vascular patency and bone appearance. Bone scintigraphy was performed for 5 rabbits in each group. On bone scintigraphic scans, the bone component of the flaps was visualized in groups 1 and 2, but not in group 3. A quantitative analysis of images was performed by drawing symmetric spherical regions of interest (ROIs) over both the implanted area and cranial bone. The uptake ratios were computed by dividing the mean counts in the implanted ROI by mean counts in the cranial bone ROI. The mean value was 0.86 +/- 0.02 in group 1 and 0.86 +/- 0.04 in group 2. A statistically significant uptake difference was not seen between venous and arterialized venous groups (P < 0.01). Histologic examination was performed all rabbits in each group, and demonstrated that the bony component was viable, showing osteocytes containing lacunae, osteoblasts along bony trabeculae, and vascular channels in groups 1 and 2. In group 3, the bony architecture of the graft was still apparent, but all bone within it was dead. There were no significant microangiographic, histologic, and scintigraphic differences between the 2 experimental methods. PMID- 15785287 TI - Hyperfibrinogenemia alone does not affect the patency of microvascular anastomosis: clinical experience and animal study. AB - Preventing vascular thrombosis in microsurgery is a prerequisite for a successful outcome. High plasma fibrinogen levels have been associated with thromboembolic risk in patients with cancer or cardiovascular disease. Patients with these comorbidities and associated hyperfibrinogenemia oftentimes require microsurgical reconstruction. This situation causes us to hesitate. Previously in our experience, 8 of 10 patients with hyperfibrinogenemia (> 500 mg/dL) underwent successful free-tissue transfer after oral cancer ablation. Based on this clinical observation, we investigated whether hyperfibrinogenemia contributes to the patency of a microvascular anastomosis. Optimal dosage of fibrinogen (300 mg/kg, intravenously) significantly increased the fibrinogen level in the plasma of the rodent hyperfibrinogenemia model. Forty male Lewis rats (weight = 300-350 g) were injected intravenously by normal saline and fibrinogen (300 mg/kg), respectively (n = 20 in each subgroup). Femoral artery and femoral vein division and reanastomosis were performed after 2 hours in rats with or without fibrinogen injection. The platelet count, prothrombin time (PT), activated partial thromboplastin time (APTT), and the platelet aggregation test induced with adenosine diphosphate (ADP) were also measured preoperatively. The ratios of circulating activated platelets as demonstrated by p-selectin (CD62P) was analyzed by flow cytometry preoperatively and 2 hours postoperatively. Laser Doppler flowmetry was used to assess the patency of the anastomosis preoperatively and 2 hours postoperatively. Vascular patency was assessed 7 days postoperatively. The results showed that the platelet count, PT and APTT levels had no significant difference among the control and the experimental group. There were no significant differences found in the ratios of CD62P expression (P = 0.65) and ADP aggregation test (P = 0.17) in comparing both groups. There were no statistical differences in the patency rates (P > 0.05) or perfusion units of femoral arteries (P = 0.84) and femoral veins (P = 0.51) after vessels division and reanastomosis, respectively. In summary, there was no correlation between experimentally induced hyperfibrinogenemia and the enhancement of thrombosis risk after microvascular surgery. This experimental data can lend support to the idea that microvascular anastomosis could be safely performed in patients with hyperfibrinogenemia alone without untoward thrombotic complications. PMID- 15785288 TI - Perioperative considerations in the autologous breast reconstruction patient with systemic lupus erythematosus. AB - Following mastectomy, women with breast cancer are faced with choices for reconstruction. In women with systemic lupus erythematosus (SLE), with or without antiphospholipid syndrome (APS), concurrent morbidities are important considerations when deciding the method of reconstruction. The 2 cases we present represent the variable spectrum of SLE and/or APS and their potential complications. When deciding the best course of action, it is most important to consider each case on an individual basis, with important emphasis on preoperative disease status. PMID- 15785289 TI - Salvage of infected cartilage grafts for nasal reconstruction with a through-and through irrigation system. AB - Development of a wound infection after nasal reconstruction can place the entire reconstructive effort in jeopardy. The approach to management in these cases has traditionally entailed wound drainage, removal of involved graft material, and debridement of nonvital tissue. Following adequate wound healing, delayed reconstruction is then performed, with the final result often compromised in form and function. We present a case of a postoperative wound infection following reconstruction of a traumatic nasal deformity utilizing autologous cartilage grafts. Treatment consisted of hospitalization with administration of culture specific parenteral antibiotics and continuous through-and-through antibiotic irrigation of the wound via an indwelling catheter. The infection was completely eradicated and all cartilage grafts were salvaged utilizing this technique. At 3 years postoperatively, the patient has maintained the shape and quality of her reconstruction, without evidence of recurrent infection. PMID- 15785291 TI - Adults and adulterations. PMID- 15785290 TI - Neuropsychological diversity in Apert syndrome: a comparison of cognitive profiles. AB - Apert syndrome is characterized by craniosynostosis, central nervous system anomalies, midface hypoplasia, and syndactyly. Current research has focused on genetic and neurologic correlates. Cognitive assessment has been primarily limited to global intellectual evaluations, which can fail to detect the diverse cognitive attributes of these children at an individual level. This report describes in detail the neuropsychological profiles of 2 children with Apert syndrome, incorporating clinical, radiographic, molecular and surgical data. One child showed intellectual deficits consistent with a moderate intellectual disability. The second child, while of normal intelligence, displayed neuropsychological deficits associated with anterior-brain-region cognitive functions. These data highlight the diversity of neuropsychological outcomes in Apert syndrome in the same genetic mutation and underline the importance of detailed neuropsychological evaluations as integral to the management protocols of affected individuals. PMID- 15785292 TI - Physical activity: the missing prescription. PMID- 15785293 TI - The Euro Cardio-QoL Project. An international study to develop a core heart disease health-related quality of life questionnaire, the HeartQoL. AB - Cardiovascular diseases, which include coronary heart disease, account for 48% of deaths in Europe and many developed countries have specifically targeted significant reductions in coronary heart disease deaths as major health care objectives. Reduction in the death rate and morbidity from coronary heart disease can be accomplished through three primary interventions: (1) modification of lifestyle behaviors; (2) use of medications; and (3) surgical procedures. In order to prescribe a specific intervention, patients with heart disease are typically diagnosed by physicians with one or more of three inter-related (but clinically distinct) conditions: myocardial infarction, angina pectoris, or heart failure. This diagnostic conceptualization of coronary heart disease has led to important improvements in specific treatments and researchers have increasingly focused their attention on comparing the efficacy of one intervention versus another. Patients, providers, and researchers have increasingly become interested in identifying those interventions that not only improve mortality but also improve symptoms, function, and health-related quality of life. Thus, researchers have developed specific symptom scales and health-related quality of life instruments for each of these three coronary heart disease conditions for use in clinical trials. As part of the Euro Cardio-QoL Project, the HeartQoL study is designed to develop a single reliable and valid core coronary heart disease specific, health-related quality of life questionnaire, to be called the HeartQoL, in order to compare outcomes with the same or across different treatments among pure or mixed populations of patients with myocardial infarction, angina pectoris, and/or heart failure. To be of value in international studies, health-related quality of life instruments must be available in a range of languages as many important clinical studies require multi-national and multi-language site collaboration. The project will recruit a total of 4200 patients with myocardial infarction (n=1400), angina (n=1400), and heart failure (n=1400) in 40 sites located in 15 countries where 13 different languages are spoken. Data will be collected using a battery of three valid self administered, health-related quality of life instruments at baseline and again within 2 to 4 weeks to develop the core HeartQoL questionnaire and to establish its reliability. Anxiety, depression, mood, personality, and generic health related quality of life will also be assessed at baseline to provide preliminary evidence of validity. PMID- 15785294 TI - Relative importance of atherosclerotic risk factors for coronary heart disease in Taiwan. AB - BACKGROUND: The relative importance of atherosclerotic risk factors, such as hypertension, dyslipidemia, diabetes and smoking, was associated with cardiovascular events and varied among different ethnic groups. For a population with relatively low coronary heart disease (CHD) such as Asian-Pacific countries, it is crucial to differentiate the roles of these risk factors. METHODS: We examined the relative importance of various risk factors for CHD in a community based cohort in Taiwan, consisting of 3602 adults aged 35 and older with a median follow-up time of 9.0 years since 1990. Regular death certificate verification and medical record reviews were performed in the follow-up activities. RESULTS: There were 85 cases defined as CHD. In the Cox proportional hazard analysis, men were at higher risk than women [hazard risk (HR)=2.22, 95% confidence interval (CI)=1.39-3.56]. Hypertension was the most common risk factor for CHD. Dyslipidemia, especially lowered high-density lipoprotein cholesterol, also played an important role (HR=2.09, 95% CI=1.33-3.29) in CHD events. Hypertension had a greater influence in males (HR=6.08, P<0.001) than in females (HR=2.80, P<0.001). No independent association was found for smoking or body mass index in cardiovascular events. CONCLUSION: This study found that in a community-based cohort, hypertension, and dyslipidemia attribute an important role to cardiovascular events. PMID- 15785295 TI - How to assess physical activity? How to assess physical fitness? AB - Regular aerobic physical activity (PA) increases exercise capacity and physical fitness (PF), which can lead to many health benefits. Accurate quantification of PA and PF becomes essential in terms of health outcome and effectiveness of intervention programmes. In this manuscript we present a review regarding the assessment of physical activity and fitness. Three types of PA assessment methods can be distinguished: criterion methods, objective methods and subjective methods. Criterion methods like doubly labelled water, indirect calorimetry and direct observation are the most reliable and valid measurements against which all other PA assessments methods should be validated, but they also hold important drawbacks. Objective PA assessment methods include activity monitors (pedometers and accelerometers) and heart rate monitoring. Finally, questionnaires and activity diaries are considered subjective methods. For the assessment of PF, we distinguish field tests and laboratory tests. The Eurofit for Adults is a test battery that is designed to assess health-related fitness of individuals, communities, sub-populations and populations. It is mainly used for evaluating the morphological component, the muscular component, the motor component and the cardio-respiratory component. In the laboratory, exercise capacity is preferentially assessed through maximal incremental exercise testing. Cardio pulmonary exercise testing is a well-established procedure that provides a wealth of clinically diagnostic and prognostic information. The peak oxygen uptake is the gold standard in the assessment of exercise tolerance. When maximal exercise is contraindicated or not achievable, the VAT or the submaximal slopes provide reasonable alternatives. PMID- 15785296 TI - Trends in blood lipid levels, blood pressure, alcohol and smoking habits from 1985 to 2002: results from INTERGENE and GOT-MONICA. AB - BACKGROUND: Favourable trends in cardiovascular disease have been observed in Sweden. The aim of this study was to study secular trends in a variety of cardiovascular risk factors. METHODS: Total-, low-density (LDL) and high-density lipoprotein (HDL) serum cholesterol; serum triglycerides; systolic and diastolic blood pressure; self-reported smoking and alcohol consumption were studied in repeated cross-sectional surveys. Data from four population-based samples in Goteborg, Sweden were used-WHO MONICA project 1985, 1990 and 1995, and INTERGENE 2002. A total of 2931 females and 2691 males aged 25-64 consisting of 1021-1624 randomly selected subjects at each survey period participated. RESULTS: Serum cholesterol levels showed downward trends but the decline in both total- and LDL cholesterol seems to be levelling off from 1995 and onwards. No significant changes were observed in serum triglyceride, HDL-serum cholesterol or blood pressure levels. The majority of the participants had higher total- and LDL-serum cholesterol levels than currently recommended. Antihypertensive medical treatment increased in women and the oldest men. The prevalence of smoking decreased from 39 to 25% in women and 35 to 20% in men respectively from 1985-2002. In contrast, the prevalence of subjects consuming strong beer and wine, respectively, at least once a week almost doubled from 1990-2002. CONCLUSIONS: Cardiovascular risk factor patterns change continuously and need to be monitored. The favourable trends in LDL-serum cholesterol and smoking in the Goteborg surveys were paralleled by less favourable trends in being overweight and alcohol consumption. PMID- 15785297 TI - Associations between estimated aerobic fitness and cardiovascular risk factors in adults with different levels of abdominal obesity. AB - BACKGROUND: We investigated the association between estimated aerobic fitness and cardiovascular risk factors, and how the association is affected by abdominal obesity. DESIGN: Cross-sectional population study. METHODS: Participants comprised 3820 adults aged 25 to 64 years from the FINRISK 2002 Study in Finland. Aerobic fitness was estimated using a non-exercise test. Waist-to-hip ratio (WHR), blood pressure, total cholesterol, high-density lipoprotein cholesterol (HDL-C), triglycerides, HDL-C to total cholesterol ratio, and gamma-glutamyl transferase (GGT) levels were measured by standardized methods. RESULTS: After controlling for age, smoking and alcohol consumption, aerobic fitness was inversely associated with systolic (P=0.027) and diastolic (P<0.001) blood pressure, total cholesterol (P=0.009), triglycerides (P=0.001), and GGT (P<0.001), and directly associated with HDL-C (P<0.001) and HDL-C to total cholesterol ratio (P<0.001) in men. In women, inverse associations were found for diastolic blood pressure (P=0.027) and triglycerides (P<0.001), and direct association for HDL-C (P<0.001) and HDL-C to total cholesterol ratio (P<0.001). Waist-to-hip ratio was independently associated with a better risk factor profile in both sexes. Interactions were found between fitness and WHR in relation to total cholesterol (P=0.001), HDL-C to total cholesterol ratio (P=0.005), triglycerides (P=0.001), and systolic (P=0.009) and diastolic (P<0.001) blood pressure among men only. CONCLUSIONS: Our data suggest that good estimated aerobic fitness is associated with a better cardiovascular risk factor profile, regardless of the level of abdominal obesity in Finnish men and women. Men in the highest WHR third seem to have more benefit of aerobic fitness on their cardiovascular risk levels than men with lower WHR. PMID- 15785298 TI - The validity of the Finnish Hospital Discharge Register and Causes of Death Register data on coronary heart disease. AB - BACKGROUND AND DESIGN: The recent introduction of sensitive markers of myocardial injury is likely to affect the epidemiology of coronary heart disease (CHD). The American Heart Association together with other societies and research agencies have recently published a new definition on acute CHD to improve consistency in epidemiological and clinical studies (referred here as the '2003 definition'). METHODS: In this study we compare the data on CHD events in the Finnish National Hospital Discharge Register (HDR) and the Causes of Death Register (CDR) with the population-based myocardial infarction (MI) register, FINMONICA/FINAMI. The FINMONICA/FINAMI events were classified according to the 2003 definition. The relevant International Classification of Diseases (ICD) codes from the HDR and CDR were used. RESULTS: Using the 2003 definition as the reference, the overall sensitivity of the ICD codes for MI in the combined HDR and CDR was 83% and the positive predictive value (PPV) was 90%. When the ICD codes for unstable angina were added to the analyses, the sensitivity improved to 85% and the PPV declined to 83%. In the age group 35-74 the sensitivity of the MI codes improved over time, in men from 64% in 1988-1992 to 81% in 1998-2002, and in women from 61 to 78%, respectively. The oldest age group, 75 years or older, had sensitivity and PPV values comparable to those of the younger. CONCLUSION: Diagnoses of fatal and non-fatal CHD events in the Finnish HDR and Causes of Death register were reasonably valid indicators for hard CHD events when compared with the FINMONICA/FINAMI register data. PMID- 15785299 TI - Quality of life and specific problems after cardiac surgery in adolescents and adults with congenital heart diseases. AB - BACKGROUND: Grown-ups with congenital heart disease (GUCH) constitute an increasing population. Some of them reach adulthood without intervention and may present with symptoms, some require first intervention or re-operation for various reasons. Although interventions become more and more frequent in these patients, limited knowledge exists on their quality of life (QoL). The aim of the present study was to analyze QoL in GUCH patients who underwent cardiac surgery after the age of 14 years. DESIGN: A total of 296 patients with a mean age of 35+/-16 years (range 14-72 years) were operated on at our institution between July 1987 and December 2000, mainly for atrial septal defect (ASD), outflow tract lesion, Marfan syndrome and co-arctation. Early mortality was 3.4%. During follow up QoL was assessed with the short form 36 health survey questionnaire (SF-36) and an additional questionnaire focused on medical and psychosocial aspects. Results were analyzed for the total collective and in relation to the underlying congenital heart disease (CHD). RESULTS: Quality of life was excellent and similar to an age- and gender-matched standard population, except in patients following repair of complete AV-canal. The main restrictions in this group were found in the emotional aspect (62.5+/-29.9) and physical role function (60.5+/ 25.0) and reflected in limitations of daily activity. CONCLUSION: Outcome and QoL in adolescents and adults with CHD is excellent and similar to that of an age- and gender-matched standard population. In the future special attention should be focused more strongly on medical follow-up and psychosocial problems in this increasing group of patients. PMID- 15785300 TI - Fibrinogen and cigarette smoking in men and women in the European Prospective Investigation into Cancer in Norfolk (EPIC-Norfolk) population. AB - BACKGROUND: Plasma fibrinogen may be an independent risk factor for cardiovascular disease. Cigarette smoking is a well-recognized determinant of plasma fibrinogen however it remains unclear how fibrinogen levels relate to the degree and duration of smoking, or to time since smoking cessation. METHODS: In a population-based study of 11 059 men and women aged 45-74 years, we examined the cross-sectional relationship between plasma fibrinogen and cigarette smoking habit. RESULTS: Mean fibrinogen concentrations were higher in current smokers compared to non-current smokers (men: 3.13+/-0.77 versus 2.80+/-0.71 g/l, P<0.0001; women: 3.03+/-0.72 versus 2.95+/-0.71 g/l, P=0.01), independent of age, body mass index and hormone replacement therapy in women In men, fibrinogen concentrations declined with years since stopping smoking but remained higher than in life-long non-smokers for 15 years. No relationship between fibrinogen and duration of smoking cessation was observed in women. On multivariate analysis, age, body mass index, use of hormone-replacement therapy, smoking status and pack-years of smoking were independent predictors of plasma fibrinogen. CONCLUSIONS: Plasma fibrinogen is strongly associated with cigarette smoking with a dose-response relationship with total pack-years of smoking. In men who stop smoking plasma fibrinogen may remain elevated for several years after cessation. PMID- 15785301 TI - The associations between physical activity, inflammation, and coagulation markers, in people with metabolic syndrome: the ATTICA study. AB - OBJECTIVE: Metabolic syndrome is a condition that promotes atherosclerosis and increases the risk of cardiovascular mortality and morbidity. We evaluated whether leisure time physical activity is associated with the levels of inflammatory and coagulation markers, in people with metabolic syndrome. METHODS: From May 2001 to December 2002 we randomly enrolled 1514 men and 1528 women (>18 years old), without any clinical evidence of cardiovascular disease, stratified by age-gender (census 2001). The population of the study was divided into those who fulfilled the NCEP ATP III criteria for the metabolic syndrome (n=701 or 33% men and 13% women) and the rest of the participants (n=2341). We assessed the relationship between self-reported physical activity status and inflammatory, and coagulation markers [i.e., C-reactive protein (CRP), serum amyloid-A (SAA), interleukin (IL)-6, tumour necrosis factor (TNF)-alpha, white blood cell (WBC) counts, and fibrinogen (FIB)], after taking into account the effect of several confounders. RESULTS: Of the non-metabolic syndrome group, 56% of men and 58% of women were classified as sedentary, while of the metabolic syndrome group 58% men and 72% women were sedentary. After controlling for various potential confounders we found that physically active individuals with the metabolic syndrome had 36% lower levels of CRP, 15% lower levels of WBC, 19% lower levels of SAA, 15% lower levels of TNF-alpha, 30% lower levels of IL-6 and 15% lower levels of FIB, compared to sedentary (all P<0.05). Similar results were observed in the non metabolic syndrome group. CONCLUSIONS: The adoption of a physically active lifestyle is independently associated with lower levels of the investigated biomarkers in individuals with the metabolic syndrome. The latter may suggest a pathway for reducing cardiovascular events, even in high-risk people. PMID- 15785302 TI - Cholesterol control in stroke prevention in Italy: a cross-sectional study in family practice. AB - BACKGROUND: Stroke represents worldwide the second and seventh cause of death and invalidity, respectively. Patients with ischaemic stroke or transitory ischaemic attack (TIA) are at high risk of recurrence, therefore requiring intensive treatment. Hypercholesterolaemia is a modifiable risk factor for stroke. The general practitioners attitude towards detection and treatment of dyslipidaemia among patients with stroke or TIA in Italy is unknown; we therefore aimed to address this issue taking advantage of the database of The Italian College of General Practitioners. METHODS: Prevalence of the monitored factors (lipid levels, statin prescription, and lipid level control with hypolipidaemic agents prescription) were analysed on a patient population of 465 061. RESULTS: A total of 2555 (49% women and 51% men) patients with a diagnosis of stroke and 2755 patients (52% women and 48% men) with a diagnosis of TIA were included in the study. Total plasma cholesterol (TC) was reported in more than 60% of the patients and low-density lipoprotein cholesterol (LDLc) and high-density lipoprotein cholesterol (HDLc) in less than half. Total plasma cholesterol and LDLc were controlled in 70.3 and 72.8% of the patients, respectively. The percentage of controlled patients decreased to 64% when both LDLc and TC were considered. Statins and fibrates were prescribed in a small proportion of patients (16.9 and 3.5%, respectively). An acceptable control of blood lipids was achieved in a majority of those patients (60.2%). However a relatively large number of patients (646) with high plasma lipids remained untreated. CONCLUSIONS: Monitoring and intervention strategies on plasma lipid levels in patients with a diagnosis of stroke or TIA need to be improved. PMID- 15785303 TI - Growth hormone-induced reduction of soluble apoptosis mediators is associated with reverse cardiac remodelling and improvement of exercise capacity in patients with idiopathic dilated cardiomyopathy. AB - OBJECTIVE: Recent experimental and clinical data indicate that abnormal inflammatory and apoptotic processes contribute to the progression of chronic heart failure (CHF). We sought to study the effects of growth hormone (GH) on circulating soluble apoptosis mediators, and to investigate whether these GH induced anti-apoptotic effects are associated with the reduction of left ventricular (LV) volumes and attenuation of exercise intolerance in idiopathic dilated cardiomyopathy (IDC) patients. METHODS: Plasma tumour necrosis factor alpha (TNF-alpha), its soluble receptors sTNFRI and sTNFRII, and plasma apoptosis mediators soluble Fas (sFas) and sFas Ligand (sFAsL) were measured (enzyme-linked immunosorbent assay) in 12 IDC patients (NYHA III; LVEF: 24+/-2%) before and after a 3-month subcutaneous administration of GH 4 IU every other day (randomized, crossover design). Peak oxygen uptake (VO2max), as well as LV volume indices, wall thickness, and end-systolic wall stress (ESWS) were also determined at the same period. RESULTS: Treatment with GH produced a significant reduction in plasma TNF-alpha (7.8+/-1.1 versus 5.5+/-0.9 pg/ml, P<0.02), sTNFRI (4.0+/-0.4 versus 3.3+/-0.3 ng/ml, P<0.05), sTNFRII (2.8+/-0.3 versus 2.4+/-0.2 ng/ml, P<0.05), sFas (4.7+/-0.7 versus 3.3+/-0.5 ng/ml, P<0.05) and sFasL (33.5+/-9.7 versus 20.2+/-6.2 pg/ml, P<0.01). A significant reduction in ESWS (841+/-62 versus 634+/-48 g/cm, P<0.01), LV end-systolic volume index (LVESVI, 128+/-12 versus 102+/-12 ml/m, P<0.001) and LV end-diastolic volume index (LVEDVI, 228+/ 16 versus 200+/-18 ml/m, P<0.01) as well as a significant increase in VO2max (15.3+/-0.7 versus 17.1+/-0.9 ml/kg per min, P<0.01) were also observed in the patient population after GH administration. Significant correlations were found between the GH-induced decrease of sTNFRII and sFasL and respective reduction of LVESVI. CONCLUSION: Growth hormone administration reduces circulating TNF system and soluble apoptosis mediators in patients with IDC. These GH-induced anti apoptotic effects may be associated with the improvement in exercise capacity as well as with the reverse of LV remodelling in patients with CHF and IDC. PMID- 15785305 TI - On the relationship between all-cause, cardiovascular, cancer and residual mortality rates with age. AB - BACKGROUND: The existence of a highly significant linear relationship between the natural logarithm (ln) all-cause mortality rate and age at the population level is firmly established (r>0.99). The slope and intercept of the equation, however, vary markedly between populations. Whether this relationship also applies to specific disease entities has not been established. METHODS: Use was made of mortality rates for all-cause, total cardiovascular, total cancer and residual diseases. The midpoint of 5-year age classes between the ages of 35 and 84 years, obtained for both sexes, were analysed. The mean of the three latest available years, from the period 1997-1999 were used. RESULTS: The relationship also applies to a slightly lesser degree to the relationship between total cardiovascular mortality rate, consisting predominantly of ischemic heart disease and stroke, and age (r>0.99). Marginally better relationships are obtained using a second-degree polynomial equation between ln all-cause mortality rate and age, age as independent variables. Total ln cancer mortality rate, however, behaves differently with a significant negative deviation of the mortality rate from linearity at older ages. Residual mortality (non-cancer, non-cardiovascular) mortality shows a mirror pattern to cancer mortality. This residual mortality expressed as a percentage of all-cause mortality varies markedly between populations. The level of some major constituents of the residual mortality rates (respiratory diseases, pneumonia, ill-defined causes and senility) also varies markedly. CONCLUSIONS: The magnitude of the variation suggests misclassification or misdiagnosis of several important disease entities, for example, between senility and stroke or between pneumonia and lung cancer. This questions the validity of disease-specific mortality rates especially at older ages, making their comparison between countries less reliable. PMID- 15785304 TI - Habitual low-intensity exercise does not protect against myocardial dysfunction after ischemia in rats. AB - BACKGROUND: It is well established that participation in a chronic exercise program can reduce coronary heart disease (CHD) risk factors and improve myocardial tolerance to ischemia-reperfusion (I-R) injury. Low-intensity exercise programs are known to be effective in reducing CHD risk factors in humans and rats, but whether similar programs are of sufficient intensity to improve intrinsic tolerance to I-R injury has not been established. Thus, the purpose of this study is to determine whether low-intensity exercise provides self protection to the heart against I-R injury. METHODS: Male, Sprague-Dawley rats were exercised on a treadmill at an intensity of 55-60% VO2max, 40 min/day, 5 days/week for 16 weeks. Reperfusion injury following 20 min of global ischemia was evaluated using the isolated perfused working heart model. Left ventricular content of the cytoprotective protein heat shock protein 70 (HSP70) was determined by Western blotting. RESULTS: The exercise program elevated HSP70 2.7 fold, but did not provide enhanced protection following 20 min of ischemia. Final post-ischemic recovery of cardiac external work was 63+/-9% of pre-ischemic value in the sedentary group (n=9) and 51+/-11% in the exercising group (n=9) (P>0.05). Post-ischemic lactate dehydrogenase release was also similar between groups and the magnitude of release was low, consistent with stunning. CONCLUSIONS: Regular exercise at 55-60% VO2max is below the threshold intensity necessary to induce intrinsic cardioprotection against I-R injury. Furthermore, elevated myocardial HSP70 is not necessarily a marker of improved protection against dysfunction associated with stunning. PMID- 15785306 TI - The relationships between plasma total homocysteine and selected atherosclerotic risk factors according to the C677T methylenetetrahydrofolate reductase gene in Japanese. AB - BACKGROUND: Variation of plasma total homocysteine (tHcy) levels among individuals is modified by 5,10-methylenetetrahydrofolate reductase (MTHFR) genotypes and selected atherosclerotic risk factors. DESIGN: Cross-sectional study. METHODS: Healthy Japanese men (n=213) and women (n=242) aged 40-69 years were examined. RESULTS: Serum uric acid, hemoglobin, hematocrit, serum creatinine, and men were independently significantly associated with plasma tHcy. The relationship between serum uric acid and plasma tHcy was the strongest in those with the mutant homozygote (VV), but the overall statistical test of interaction was not significant. CONCLUSIONS: Serum uric acid, hematocrit, hemoglobin, creatinine, and men were determinants of plasma tHcy levels. Further investigations are needed. PMID- 15785307 TI - Relationship between paraoxonase-1 (PON1) activity and lipoprotein (a) levels in Turkish coronary artery disease patients living in the Antalya region. AB - BACKGROUND: Turkish adults have a high incidence of coronary artery disease (CAD). DESIGN: The role of lipoprotein(a) (Lp[a]) as a risk factor for CAD and its association with paraoxonase-1 (PON1) activity and lipid peroxidation indicators were evaluated in Turkish subjects living in Antalya. METHODS: Paraoxonase-1 activity, Lp(a), conjugated dienes (CD) and thiobarbituric acid reactive substances (TBARS) levels were measured in 45 healthy controls and 72 CAD patients. RESULTS: In CAD patients, decreased PON1 activity and increased Lp(a), CD and TBARS levels were observed. Risk factors associated with CAD included TBARS, age and CD. CONCLUSIONS: Our results indicate an association between PON1 activity and Lp(a) level in CAD patients. PMID- 15785309 TI - Mobile phones and cancer? PMID- 15785310 TI - Behaviour and motives of adolescents towards skin cancer. PMID- 15785311 TI - Epidemiology of cancer in children under one year of age in Mexico City. AB - In this work, the epidemiology of cancer in children under one year of age in Mexico City is described. A survey (observational, descriptive and prolective study) from 1 January 1996 to 31 December 1999 was conducted at two paediatric hospitals of the Instituto Mexicano de Seguro Social in Mexico City (IMSS-MC). To calculate both the general and the by sex incidence (rates are given per 10) all new cases recorded for children under one year of age (numerator) and Mexico City population served by the IMSS (denominator) were used. When the total of 34 cases that fulfilled the requirements was analysed, an incidence of 194.5 was obtained. Leukaemia occupied first place with a rate of 68.6 and hepatic and germinal cells tumours occupied second place with an incidence of 28.6, whereas peripheral nervous system tumours (neuroblastoma) showed a very low rate (11.4). Overall, the male/female ratio for tumours was 1.4:1, with the ratio varying with different types of tumours. Cancer incidence in this population was shown to be close to that in developed countries, but differed in the distribution of the type of tumour: leukaemia had a very high incidence and that for neuroblastoma was very low. PMID- 15785312 TI - Survival patterns for the top four cancers in Canada: the effects of age, region and period. AB - This study examined the variations in survival rates (1989-1991) and the trends (1969-1991), by sex, age and province, for patients diagnosed with breast, colorectal, lung or prostate cancer in Canada and compared the Canadian rates with those of nine American SEER registries. Five-year age-standardized relative survival rates (ASRs) were calculated, and the trends were estimated from variance-weighted linear regression of the ASRs for five periods of diagnosis (1969-1973, 1974-1978, 1979-1983, 1984-1988 and 1989-1991). In 1989-1991, the ASR varied among provinces for each cancer except female colorectal cancer. The lowest survival rates were observed in the youngest patients (15-44) for breast and prostate cancers, and in the oldest patients (75-99) of both sexes for lung and colorectal cancers. Over the five periods, a major trend toward improved survival was observed for breast, prostate and colorectal cancers (P<0.008), whereas no changes were seen for lung cancer. The ASRs in the western region were higher than in the Atlantic region over time (P<0.02) for each cancer. From the third period onward, the ASRs for Canadian patients with lung cancer were similar to those for the US patients and lower than for Canadian patients with breast, prostate or colorectal cancer. The observed increases in ASR for breast and prostate cancer are likely due to the increased use of screenings and the improved treatment modalities. PMID- 15785313 TI - Modifying effects of Terminalia catappa on azoxymethane-induced colon carcinogenesis in male F344 rats. AB - The modifying effects of dietary administration of an herb, Terminalia catappa (TC), were investigated on rat colon carcinogenesis induced by a carcinogen azoxymethane (AOM). The number of aberrant crypt foci (ACF) and beta-catenin accumulated crypts (BCACs) in the colon, and proliferating cell nuclear antigen (PCNA) labelling index in the colonic epithelium were examined in a total of 36 male F344 rats. All animals were randomly divided into five experimental groups (4-10 rats in each group). At 6 weeks of age, rats in groups 1, 2 and 3 were given s.c. injections of AOM once a week for 2 weeks at a concentration of 20 mg/kg body weight. One week before the first injection of AOM, rats in groups 2 and 3 were fed a diet containing 0.02 and 0.1% TC, respectively, throughout the experiment. Rats in group 4 were fed a diet containing 0.1% TC. Rats in group 5 were served as untreated controls. All animals were sacrificed at the experimental week 5 after the start of the experiment. Oral administration of TC at both doses significantly decreased the numbers of both ACF/colon/rat (P<0.05 for 0.02% TC, P<0.005 for 0.1% TC) and BCAC/cm/rat (P<0.05 for both 0.02 and 0.1% TC), when compared with the control group (group 1). Colonic PCNA labelling index in groups 2 and 3 was also significantly lower than that in group 1 (P<0.001 for 0.02% TC, P<0.005 for 0.1% TC). These results suggest that TC has a potent short term chemopreventive effect on biomarkers of colon carcinogenesis and this effect may be associated with the inhibition of the development of ACF and BCACs. PMID- 15785314 TI - Comparison of early performance indicators for screening projects within the European Breast Cancer Network: 1989-2000. AB - In 1989 the European Breast Cancer Network (EBCN) was established by the first pilot projects for breast cancer screening, co-funded by the Europe Against Cancer programme. We report early performance indicators for these EBCN projects while taking into account their organizational setting. Out of 17 projects in the network, 10 projects from six European countries contributed aggregated data on number of invitations, screening examinations, and breast cancers detected over the period 1989-2000. Results were summarized separately for projects in centralized versus decentralized health care environments. The European Guidelines for quality assurance in mammography screening provided reference values for the performance indicators. The most prominent finding in this study was the higher participation rate in centralized versus decentralized projects (average participation in 1998: 74 versus 33%; P<0.001), whereas the invitation system and screening policy in these projects were similar. Detection rates and characteristics of cancers detected at initial and subsequent screening examinations showed no significant differences between centralized and decentralized projects. Even though early performance indicators for centralized versus decentralized projects were similar, the impact of breast screening on mortality from this disease at the population level will differ since the decentralized projects reach only part of the target population. PMID- 15785315 TI - The influence of mammographic screening on national trends in breast cancer incidence. AB - Introducing an organized mammographic screening programme affects the breast cancer incidence rate in a population. The diagnosis is advanced in time, and initially, an increase will occur in the number of cases, followed by a drop in the rate when women leave the programme. The aim of this study was to quantify the potential effects that mammographic screening programmes have on breast cancer incidence. In addition, we wanted to investigate how the incidence of breast cancer varies between different birth cohorts, age groups and time periods in the five Nordic countries Finland, Denmark, Iceland, Norway and Sweden, adjusting for the effects of the screening programmes. Time trends were analysed over the period 1978-1997, using age-period-cohort models. In Sweden, the rates more than doubled (relative risk (RR)=2.20, 95% confidence interval (CI) 1.8-2.6) in women offered screening for the first time compared with women not offered screening. The risk remained elevated (RR=1.34, 95% CI 1.2-1.6) for women who were continued to be offered screening, compared with women who were not offered screening. Finally, the rates dropped (RR=0.68, 95% CI 0.6-0.8) when the women left the programme. This indicates that screening advances the time of diagnosis, which is a prerequisite to subsequent reduction in mortality. Analysis of secular trends, corrected for the influence of screening, showed that the rates in Finland increased by 13% per 5-year period, with a more modest increase in the other countries. There were strong cohort effects in all Nordic countries, and the risk seemed to be flattening for the youngest cohorts in most of the countries. PMID- 15785316 TI - Parity-induced changes in global gene expression in the human mammary gland. AB - The protective effect of an early first full-term pregnancy in relation to breast cancer risk is well established, but the molecular and cell-specific changes in the human mammary gland involved remain unclear. To identify the molecular changes associated with pregnancy-induced differentiation, we analysed the global gene expression profiles of normal mammary tissues from both a parous and a nulliparous woman, using serial analysis of gene expression. This approach allowed us to identify sets of genes, known and unknown, that are differentially expressed in parous versus age-matched nulliparous mammary gland tissues. The normal mammary gland of a multiparous woman is characterized by several known differentiation markers such as casein kappa, casein beta, keratin 14, CCAAT/enhancer binding protein beta and delta and adipsin. Candidate genes involved in cytoarchitectural remodelling and growth inhibition with a potential role in pregnancy-induced protection against breast cancer were also observed. Several genes that are highly expressed in the nulliparous mammary gland and that are lost after pregnancy, encode for growth promoting, cytoskeletal and extracellular matrix proteins. One of these genes, the small breast epithelial mucin, is almost completely downregulated upon a first full-term pregnancy but is known to be expressed in more than 90% of invasive ductal carcinomas. PMID- 15785317 TI - Resveratrol and breast cancer risk. AB - Resveratrol is a non-flavonoid polyphenol that has attracted attention as a potential anticancer agent in vitro and in vivo, but scanty epidemiological data are available. We have therefore analysed the relation between dietary intake of resveratrol and breast cancer risk using data from a case-control study conducted between 1993 and 2003 in the Swiss Canton of Vaud on 369 cases and 602 controls. Compared with the lowest tertile of total resveratrol intake, the multivariate odds ratios (OR) were 0.50 for the intermediate and 0.39 for the highest tertile, and the trend in risk was significant. A significant inverse association was observed for resveratrol from grapes (OR = 0.64 and 0.55), but not for wine. The inverse relation between resveratrol and breast cancer risk was not explained by several potential confounding factors, including detailed allowance for alcohol intake, nor attributable to a non-specific favourable effect of fruit on breast cancer risk. PMID- 15785318 TI - NOD2 variants and the risk of malignant melanoma. AB - Both hereditary and environmental factors are important in the aetiology of malignant melanoma. Among the risk factors for malignant melanoma are immunodeficiency and immunosuppression. The recently identified NOD2 gene is involved in the regulation of immune function through activation of the transcription factor nuclear factor-kappaB (NF-kappaB). Three common NOD2 mutations -- 3020insC, G908R and R702W -- have been shown to be associated with chronic inflammatory disease such as Crohn's disease, the 3020insC also with human malignancy colorectal cancer. We examined the frequency of the NOD2 variants in 424 patients with malignant melanoma and 649 controls. The 3020insC mutation was present in 6.9% of unselected cases and 7% of the controls (odds ratio (OR) 1.0; P not significant). The mutation was present in 6.8% of 162 cases diagnosed under the age of 50 and in 7.1% of cases diagnosed after the age of 50. A mutation was present in the index case in 5% of 40 familial melanomas (OR 0.7; P not significant). There were no statistically significant differences between prevalence of G908R and R702W in malignant melanoma patients and controls. In conclusion, the three common NOD2 mutations are not associated with increased risk of development of malignant melanoma. PMID- 15785319 TI - Ecological association between indoor radon concentration and childhood leukaemia incidence in France, 1990-1998. AB - The objective of this study was to evaluate the ecological association between indoor radon concentration and acute leukaemia incidence among children under 15 years of age in the 348 geographical units (zones d'emploi, ZE) of France between 1990 and 1998. During that period, 4015 cases were registered by the French National Registry of Childhood Leukaemia and Lymphoma. Exposure assessment was based on a campaign of 13 240 measurements covering the whole country. The arithmetic mean radon concentration was 85 Bq/m (range, 15-387 Bq/m) and the geometric mean, 59 Bq/m (range: 13-228 Bq/m). A positive ecological association, on the borderline of statistical significance (P=0.053), was observed between indoor radon concentration and childhood leukaemia incidence. The association was highly significant for acute myeloid leukaemia (AML) (P=0.004) but not for acute lymphocytic leukaemia (ALL) (P=0.49). The standardized incidence ratio (SIR) increased by 7, 3 and 24% for all acute leukaemia, ALL and AML, respectively, when radon concentration increased by 100 Bq/m. In conclusion, the present ecological study supports the hypothesis of a moderate association between indoor radon concentration and childhood acute myeloid leukaemia. It is consistent with most previous ecological studies. Since the association is moderate, this result does not appear inconsistent with the five published case-control studies, most of which found no significant association. PMID- 15785320 TI - Inhibition of two stage renal carcinogenesis, oxidative damage and hyperproliferative response by Nigella sativa. AB - In an earlier communication we reported that Nigella sativa suppresses potassium bromate-induced renal oxidative damage. In the present study, we report the chemopreventive effect of Nigella sativa against ferric nitrilotriacetate (Fe NTA)-induced renal oxidative stress, hyperproliferative response and renal carcinogenesis. Fe-NTA (9 mg Fe/kg body weight, intraperitoneally) enhances renal lipid peroxidation, xanthine oxidase, gamma-glutamyl transpeptidase and hydrogen peroxide (H2O2) generation with reduction in renal glutathione content, antioxidant enzymes and phase II metabolizing enzymes. It also caused increase in blood urea nitrogen, serum creatinine, ornithine decarboxylase (ODC) activity and thymidine [H] incorporation into renal DNA. It also enhanced DEN (N diethylnitrosamine)-initiated renal carcinogenesis by increasing the percentage incidence of tumours. Treatment of rats orally with Nigella sativa (50 and 100 mg/kg body weight) resulted in significant decrease in gamma-glutamyl transpeptidase, lipid peroxidation, xanthine oxidase, H2O2 generation, blood urea nitrogen, serum creatinine, renal ODC activity, DNA synthesis (P<0.001) and incidence of tumours. Renal glutathione content (P<0.01), glutathione metabolizing enzymes (P<0.001) and antioxidant enzymes were also recovered to significant levels (P<0.001). Thus, our data suggest that Nigella sativa is a potent chemopreventive agent and suppresses Fe-NTA-induced oxidative stress, hyperproliferative response and renal carcinogenesis in Wistar rats. PMID- 15785321 TI - Alcohol consumption and the risk of cancer in Japanese men: the Miyagi cohort study. AB - The objective of this study was to investigate the association between alcohol consumption and the risk of total cancer, and to estimate the proportion of total cancer attributable to drinking habit in Japanese men. From June through August 1990, a total of 21 201 Japanese men completed a self-administered questionnaire on various health habits, including alcohol consumption. During 153 389 person years of follow-up through December 1997, we identified a total of 882 cases of cancer. We used Cox proportional hazards regression to estimate the relative risk of total cancer according to categories of alcohol consumption. The risk for total cancer was significantly higher in ex-drinkers than never-drinkers. There was a dose-response relationship between the amount of alcohol consumed and the risk of total cancer among current drinkers: multivariate RRs in reference to never-drinkers (95% confidence intervals (CI)) were 1.1 (0.8-1.3), 1.3 (1.0-1.7), and 1.3 (1.1-1.7) in current drinkers who consumed less than 22.8 g, 22.8-45.5 g, 45.6 g or more alcohol per day, respectively (P for trend <0.001). Estimated 17.9% (95% CI 3.1-30.5) of total cancer risk was attributable to drinking habit. In our findings, approximately 20% of the total cancer cases in Japanese men may be prevented by alcohol control. PMID- 15785322 TI - Genotoxicity evaluation of electromagnetic fields generated by 835-MHz mobile phone frequency band. AB - It is still unclear whether the exposure to electromagnetic fields (EMFs) generated by mobile phone radiation is directly linked to cancer. We examined the biological effects of an EMF at 835 MHz, the most widely used communication frequency band in Korean CDMA mobile phone networks, on bacterial reverse mutation (Ames assay) and DNA stability (in vitro DNA degradation). In the Ames assay, tester strains alone or combined with positive mutagen were applied in an artificial mobile phone frequency EMF generator with continuous waveform at a specific absorption rate (SAR) of 4 W/kg for 48 h. In the presence of the 835-MHz EMF radiation, incubation with positive mutagen 4-nitroquinoline-1-oxide and cumene hydroxide further increased the mutation rate in Escherichia coli WP2 and TA102, respectively, while the contrary results in Salmonella typhimurium TA98 and TA1535 treated with 4-nitroquinoline-1-oxide and sodium azide, respectively, were shown as antimutagenic. However, these mutagenic or co-mutagenic effects of 835-MHz radiation were not significantly repeated in other relevant strains with same mutation type. In the DNA degradation test, the exposure to 835-MHz EMF did not change the rate of degradation observed using plasmid pBluescript SK(+) as an indicator. Thus, we suggest that 835-MHz EMF under the conditions of our study neither affected the reverse mutation frequency nor accelerated DNA degradation in vitro. PMID- 15785324 TI - The intrauterine origin of male breast cancer: a birth order study in Denmark. AB - First pregnancies are known to have higher oestrogen levels than later ones and first-born women are at increased breast cancer risk compared with later-born women. We hypothesized that a birth order effect might be even more evident in male breast cancer patients, in whom oestrogens in adult life are generally low. In a population-based study in Denmark involving 77 male breast cancer patients and 288 population controls, first-born men compared with later-born men had a relative risk of 1.71 for the disease (95% confidence interval (CI) 1.00-2.92). This result is in line with that seen in female breast cancer cases and indicates that male breast cancer may have roots in the intrauterine life, oestrogens being a likely mediator. PMID- 15785323 TI - A pilot study of participation in faecal occult blood testing and screening colonoscopy after health education in Hong Kong. AB - Colorectal cancer is the second commonest cancer in Hong Kong. The screening behaviour of the Chinese population has not been assessed. The aim of this study is to report a pilot study of educating and subsequent evaluation of colorectal cancer screening behaviour in the Hong Kong Chinese population. Subjects were invited to attend a free health talk on colorectal cancer. Both self-paid faecal occult blood testing (FOBT) and free screening colonoscopy were offered after the education programme. Of the participants taking part in the education programme 113/119 (95%) completed the FOBT. Of the FOBT 8/113 (7%) showed positive result and three patients had neoplasia at colonoscopy. Twenty-five patients with negative FOBT also completed colonoscopy; two had adenomas. Screening colonoscopy after FOBT was accepted by 28% of subjects. Those younger than 65 years and those with a positive FOBT (7/8 versus 25/105 for those with negative FOBT, P=0.0003) were more likely to agree to screening colonoscopy. In conclusion, health education is important for ensuring high acceptance and implementation of colorectal cancer screening in Hong Kong Chinese. FOBT is an acceptable and feasible screening method in Hong Kong. PMID- 15785325 TI - Lung cancer in Africans in a South African city population in transition. AB - In South Africa, a study has been carried out on a series of African patients with lung cancer, who were admitted in 1994-1999 to King Edward VIII Hospital (2000 beds), and who lived in Durban (population includes approximately 650 000 resident Africans). In the urban context, Africans have recently experienced numerous changes in environmental, socio-economic and dietary factors and in other respects, thereby undergoing considerable westernization of lifestyle, compared with the past and the situation still prevailing among Africans in rural areas. In the period indicated, the mean annual number of patients admitted with lung cancer included 56 men and 9 women, thereby yielding standardized incidence rates of 15.5 and 3.5, respectively, per 100 000. While such data have their limitations, it is noteworthy that in the South African Cancer Registry for 1993 1995 the corresponding rates (pathology based) for African men and women for the whole country, rural and urban, were 11.7 and 2.6, respectively, per 100 000. In comparison, in the Registry the corresponding rates reported for white South African men and women were 22.3 and 12.3 per 100 000. With the ongoing transitional changes prevailing, most particularly concerning rises in smoking among adolescents, there is little chance of controlling further rises in the occurrence of the disease. It is significant that in the US, African Americans have been reported to have the highest rate for lung cancer in the world. PMID- 15785326 TI - Hair-raising science. PMID- 15785327 TI - More research on facial hair. PMID- 15785329 TI - Executive summary: The State of the Science on Safe Medication Administration symposium. PMID- 15785330 TI - Home infusion therapy: essential characteristics of a successful education process: grounded theory study. AB - Little research has been published on the education process from the perspective of family caregivers. This article reports the findings from a grounded theory study of caregivers who successfully learned how to deliver home infusion therapy for a family member. The participant sample was drawn from the home infusion service of a metropolitan health maintenance organization in the western United States. The objective of the study was to identify, from the caregiver's point of view, the characteristics of an education process that resulted in the learner's competence with home infusion therapy. The knowledge gained from this study can be used to develop a framework for examining and planning practice guidelines that reflect the expertise of the nurse and the experiences of the learner. PMID- 15785331 TI - Liver function tests: their role in the diagnosis of hepatobiliary diseases. AB - Liver diseases are common, and currently represent the 12th leading cause of death in the United States. However, numerous hepatic disorders exist, and differential diagnosis often is difficult. Moreover, because laboratory testing is routine, an abnormal serum transaminase or alkaline phosphatase in patients without clinical symptoms is not uncommon. Although liver function tests are critical in recognizing the presence of liver disease and its specific diagnosis, the interpretation of the tests may be confusing and difficult. Furthermore, not all persons with one or more test abnormalities actually have liver disease. In this review, liver function tests and an approach to their interpretation are discussed. PMID- 15785333 TI - Hypodermoclysis: an alternate method for rehydration in long-term care. AB - The hypodermoclysis technique of subcutaneous infusion has many benefits for long term care patients and staff. Minor complications associated with the procedure are easily remedied, and studies have proved its effectiveness. Hypodermoclysis provides an easy-to-use, safe, and cost-effective alternative to intravenous hydration for the elderly long-term care patient. PMID- 15785332 TI - Outpatient management of electrolyte imbalances associated with anorexia nervosa and bulimia nervosa. AB - Bulimia nervosa and anorexia nervosa are eating disorders with significant morbidity that often go undetected. Nurses and primary care providers are encouraged to recognize the early signs and symptoms of these disorders and to intervene appropriately. Several case reports in this article describe patients with these disorders and various related electrolyte abnormalities. Understanding electrolyte imbalances associated with both disorders may lead to earlier effective intervention and overall improved health outcomes. PMID- 15785337 TI - The clinical orientation manual: a student/preceptor educational resource. PMID- 15785334 TI - Neonatal necrotizing enterocolitis: diagnosis, management, and pathogenesis. AB - The diagnosis of neonatal necrotizing enterocolitis is one of great concern to pediatric and neonatal clinicians. Intravenous access remains an integral part of the medical and surgical management of infants with this diagnosis, and the infusion nurse is intimately involved in the care of these patients. This article discusses the definition of necrotizing enterocolitis, presents current knowledge regarding its basic pathophysiology, and identifies common and rare sequelae of this oftentimes devastating disease of premature infants. Medical and surgical management goals of therapy are described. This overview will aid the infusion nurse in caring for these patients. PMID- 15785338 TI - Quick assessment of computer skills: setting the bar. PMID- 15785339 TI - Peer-to-peer mentoring: teaching collegiality. AB - Developing undergraduate educational initiatives that employ mentoring and encourage collegiality will assist novice nurses in seeking these relationships when they enter practice. The author describes a peer-to-peer mentoring program that used electronic mentoring to link junior and senior nursing students. Structure, process, and outcome elements of the program are discussed. PMID- 15785340 TI - The collaborative track option for BSN education: the best of both worlds. AB - As the healthcare needs of patients in acute and community settings become increasingly complex, the demand for nurses with baccalaureate and higher degrees will increase. The collaborative efforts of faculty from 1 university and 4 community colleges created a model for increasing the number of BSN graduates in 4 years. The authors describe this collaborative track model that efficiently uses the strengths and resources of both community colleges and a university for preparing baccalaureate degree nurses. PMID- 15785341 TI - Assigning readings: faculty perceptions and strategies. AB - Nursing faculty routinely assign readings to undergraduate, prelicensure students. When asked what types of readings they required, almost all said they required readings from textbooks. Most expect students to either read or skim all the assignments before class. However, experience has taught faculty that often students do not read. Strategies are provided to enhance reading experiences or provide meaningful alternatives. PMID- 15785342 TI - A profile of accelerated second bachelor's degree nursing students. AB - With economic uncertainty and the worsening of the nursing shortage, interest in accelerated second bachelor's degree programs has skyrocketed. Despite dramatic increases in the number of programs, little has been published about the characteristics of these students. A descriptive study at a mid-Atlantic public comprehensive university examined demographic characteristics and compared academic performance of students in the accelerated and traditional baccalaureate programs. Data and their implications for recruitment, admission, and curricular design are presented. PMID- 15785343 TI - Measuring learning, student engagement, and program effectiveness: a strategic process. AB - What if there was an effective way to address the age-old question from students, "Why do we have to do this assignment?" And from faculty, "How do we know our students are really learning?" And from administrators, "How will we demonstrate to our peers, our accrediting agencies, and other program stakeholders that our programs are educationally effective?" As it undertook a curriculum redesign, faculty in a baccalaureate school of nursing developed a 9-step process for curriculum implementation. The authors discuss how they applied the 9 steps strategically, positioning the program for 2 successful accreditation self studies and concurrently addressing, with greater confidence, some of these age old questions. PMID- 15785345 TI - Academic and clinical dissonance in nursing education: are we guilty of failure to rescue? AB - The experience of cognitive dissonance in novice clinical nursing students is examined. These students often confront an incongruity between the rule-bound academic ideal of nursing with which they have been prepared and the more flexible, intuition-driven clinical reality they encounter. Without insightful guidance from clinical faculty, the students' response to this dissonance could include disillusionment with clinical nursing practice or devaluation of the academic ideal of nursing. Cognitive Dissonance Theory, the Novice to Expert Model, and the Neuman Systems Model provide insight into this phenomenon and serve as a theoretical foundation for recommended strategies and interventions for optimal response to dissonance between academic ideal and clinical reality in nursing students. PMID- 15785344 TI - Don't let the grievance process cause grief. AB - Faculty in a division of nursing at a state university has developed a unique way to handle student grievances. Community nurses serve on arbitration boards. These boards hear relevant evidence from both sides and render decisions, which are binding within the division. The process has been successful and has helped take "grief" out of the grievance process. PMID- 15785346 TI - Capturing the curriculum: a curriculum maturation and transformation process. AB - The author discusses an approach to successful curriculum revision that provides faculty with a renewed sense of individual and collective ownership of curriculum change. The framework for curriculum revision includes the components of commitment, change, collaboration, collegiality, consensus, communication, closure, and celebration, and the processes used to actualize these concepts. PMID- 15785347 TI - A mental health nursing clinical experience with hospice patients. AB - The author describes a clinical experience with hospice patients developed for an 8-week mental health nursing course. The experience focused on the promotion of mental health in the patient and the therapeutic use of self with students. Students were assigned a hospice patient each of whom they visited once a week. Students kept a journal and met for a weekly support group meeting with their instructor. The experience was evaluated positively by students who thought it improved their use of therapeutic communication, empathy, and gained insight into death and dying. PMID- 15785349 TI - Aspire to excellence. PMID- 15785348 TI - Plastic surgical nursing: write about how you care for people. PMID- 15785350 TI - Self-esteem and cosmetic enhancement. PMID- 15785351 TI - Perioperative nutrition and nutritional supplements. AB - Due to the combination of poor dietary habits and aging in the U.S. population, an increase in the incidence of chronic disease is occurring in this country. The prevalence of poor nutrition, obesity, diabetes, and other chronic diseases can have a significant impact on surgical outcome and complications. The number of patients with pre-existing nutritional deficiencies and health problems who are seeking cosmetic surgical procedures is growing. Nutritional intervention and supplements can have a beneficial effect on such patients who are undergoing surgery. Plastic surgeons performing aesthetic procedures may want to incorporate nutritional guidance and supplementation into their perioperative office regimen to mitigate complications and to optimize surgical outcome. PMID- 15785353 TI - Acne vulgaris. PMID- 15785354 TI - Can a nurse witness a surgical consent form before the anesthetist has seen the patient? PMID- 15785355 TI - Introduction to the wound care department. PMID- 15785357 TI - The return of silicone gel-filled breast implants: will you be ready? PMID- 15785356 TI - Introduction to the Legal Department. PMID- 15785358 TI - Surfing the Internet on nonsurgical rejuvenation: focus on lasers. PMID- 15785360 TI - Care of the live kidney donor: consensus on the ultimate gift. PMID- 15785361 TI - A Report of the Amsterdam Forum On the Care of the Live Kidney Donor: Data and Medical Guidelines. AB - Kidney transplant physicians and surgeons met in Amsterdam, The Netherlands, from April 1-4, 2004 for the International Forum on the Care of the Live Kidney Donor. Forum participants included over 100 experts and leaders in transplantation representing more than 40 countries from around the world, including participants from the following continents: Africa, Asia, Australia, Europe, North America, and South America. PMID- 15785362 TI - Posttransplantation bone disease. AB - Transplanted patients experience rapid loss of bone, high fracture rates, and increases in morbidity and mortality as a consequence of a posttransplant scenario that is highly deleterious to the skeleton. Immune suppressive drugs, especially glucocorticoids, are toxic to bone, often acting on a background of preexisting osteodystrophy resulting from long-standing renal, hepatic, cardiac, or pulmonary disease. Cyclosporin and tacrolimus lead to a severe osteopenic state in rats, but the skeletal toxicity of the calcineurin inhibitors in the clinical environment is less clear. Nor is it clear whether cyclosporin and tacrolimus differ in their skeletal actions. Mycophenolate mofetil and sirolimus do not appear to have important skeletal toxicity. Preventative strategies include minimizing glucocorticoid exposure and implementing therapies to counter the increase in bone resorption and decrease in bone formation that follows transplantation. Antiresorptive agents, especially bisphosphonates, appear capable of retarding or halting the early bone loss and possibly reduce fracture rates also. Vitamin D and calcium are ineffective, but calcitriol has utility in some reports. Bone anabolic agents, such as synthetic parathyroid hormone and growth hormone, have potential, but data are lacking. PMID- 15785363 TI - Human organ and tissue transplantation. PMID- 15785364 TI - Commentary: World Health Assembly resolution 57.18 on xenotransplantation. PMID- 15785365 TI - Commentary: harmonizing the regulators. PMID- 15785366 TI - Commentary: the WHO resolution on human organ and tissue transplantation. PMID- 15785367 TI - The World Health Organization resolution on human organ transplantation: will it result in action? PMID- 15785368 TI - Regulatory T cells in transplantation: natural or induced? PMID- 15785369 TI - Raising the direction signposts that guide T cell trafficking into allografts. PMID- 15785370 TI - Critical influence of natural regulatory CD25+ T cells on the fate of allografts in the absence of immunosuppression. AB - BACKGROUND: Allografts are occasionally accepted in the absence of immunosuppression. Because naturally occurring CD4(+)CD25(+) regulatory T cells (natural CD25(+) Treg cells) have been shown to inhibit allograft rejection, we investigated their influence on the outcome of allografts in nonimmunosuppressed mouse recipients. METHODS: We compared survival times of male CBA/Ca skin grafts in female CBA/Ca recipients expressing a transgenic anti-HY T-cell receptor on a RAG-1(+/+) (A1[M]RAG+) or a RAG-1(-/-) (A1[M]RAG-) background. Depletion of natural CD25(+) Treg cells in A1[M]RAG+ mice was achieved by in vivo administration of the PC61 monoclonal antibody. The influence of natural CD25(+) Treg cells on the fate of major histocompatibility complex class II-mismatched (C57BL/6X bm12)F1 skin or bm12 heart transplants in C57BL/6 recipients was also assessed. Finally, we investigated the impact of natural CD25(+) Treg cells on the production of T-helper (Th)1 and Th2 cytokines in mixed lymphocyte cultures between C57BL/6 CD4(+) CD25(-) T cells as responders and bm12 or (C57BL/6X bm12)F1 antigen-presenting cells as stimulators. RESULTS: Male allografts were spontaneously accepted by female A1(M)RAG+ mice but readily rejected by female A1(M)RAG+ mice depleted of natural CD25(+) Treg cells by pretreatment with the PC61 monoclonal antibody. Depletion of CD25(+) Treg cells also enhanced eosinophil-determined rejection of (C57BL/6X bm12)F1 skin grafts or bm12 cardiac grafts in C57BL/6 recipients. Finally, natural CD25(+) Treg cells inhibited the production of interleukin (IL)-2, interferon-gamma, IL-5, and IL-13 in mixed lymphocyte culture in a dose-dependent manner. CONCLUSION: Natural CD25(+) Treg cells control Th1- and Th2-type allohelper T-cell responses and thereby influence the fate of allografts in nonimmunosuppressed recipients. PMID- 15785371 TI - Inhibition of matrix metalloproteinases during chronic allograft nephropathy in rats. AB - BACKGROUND: Chronic allograft nephropathy (CAN) belongs to the major causes of long-term kidney allograft failure. One of the histologic hallmarks of CAN is interstitial fibrosis, influenced by matrix metalloproteinases (MMPs) that are controlling extracellular matrix (ECM) degradation. Whether MMPs affect the development and progression of CAN is not clear so far. To analyze the role of MMPs in CAN, we investigated the effects of an early and a late application of BAY 12-9566, an inhibitor of MMP-2, -3, and -9 on the development and progression of CAN in a rat kidney-transplantation model. METHODS: Fisher kidneys were orthotopically transplanted into Lewis recipients that were treated with BAY 12 9566 (15 mg/kg per day) or vehicle either for the first 10 days after transplantation (early treatment) or from week 12 to week 20 after transplantation (late treatment). Proteinuria was analyzed every 4 weeks up to week 20 after transplantation when kidney grafts were removed for further analysis. RESULTS: Early MMP-inhibition resulted in a significantly reduced 24 hour protein excretion that was paralleled by a lower grade of CAN after 20 weeks. However, late MMP inhibition starting at week 12 after transplantation resulted in significantly higher proteinuria and a higher grade of CAN as compared with controls. Furthermore, transforming growth factor-beta and platelet derived growth factor-B chain mRNA levels were significantly increased in these animals. CONCLUSIONS: Inhibition of MMPs early after transplantation reduced the development and progression of CAN but promoted CAN if initiated at later stages. Thus, MMPs are involved in the development and progression of CAN. PMID- 15785373 TI - Renal transplantation: examination of the regulation of chemokine binding during acute rejection. AB - BACKGROUND: Chemokines recruit leukocytes during allograft rejection. It is thought that the formation of glycosaminoglycan (GAG)-stabilized chemokine concentration gradients within the allograft plays a crucial role in this process. This raises the possibility that changes in GAG biology might regulate chemokine binding and the development of rejection. METHODS: Immunocytochemical techniques were used to quantify changes in GAG expression within normal and rejection renal biopsy sections. Changes in GAG expression by cultured endothelial cell lines were also examined after stimulation with tumor necrosis factor-alpha and interferon-gamma. Quantitative reverse-transcriptase polymerase chain reaction was used to examine the basis for increased sulphation of heparan sulphate (HS) observed during inflammation. A binding assay was developed to determine how levels of GAG expression correlate with changes in chemokine (CCL5) sequestration. RESULTS: In normal kidney, HS was largely restricted to the tubular basement membrane; chondroitin-4-sulphate and chondroitin-6-sulphate were expressed within the interstitial tissues. The expression of all three GAGs was increased significantly during acute rejection, and heavily sulphated HS remained predominant within the tubular basement membrane. Treatment of endothelial cells with proinflammatory cytokines increased the expression of mRNA encoding N deacetylase/N-sulphotransferase-1, an isoform of the enzyme responsible for N sulphation of HS. Cytokine-treated cells and rejection biopsy specimens showed an enhanced capacity to bind CCL5. CONCLUSIONS: Chemokine production is known to be increased during acute renal allograft rejection. In this study we showed that the graft tissues also respond by increasing their potential to bind chemokines, a process that is vital for effective chemokine presentation and leukocyte recruitment. PMID- 15785372 TI - Hyperacute lung rejection in the pig-to-human model 4: evidence for complement and antibody independent mechanisms. AB - BACKGROUND: We assessed whether the combination of complement regulation and depletion of xenoreactive antibodies improves the outcome of pulmonary xenografts compared with either strategy alone. METHODS: Lungs from pigs heterozygous (hDAF(+/-)) or homozygous (hDAF(+/+)) for the human decay accelerating factor transgene (hDAF) or their nontransgenic litter mates (hDAF(-/-)) were perfused with heparinized whole human blood. In additional groups, xenoreactive natural antibodies (XNA) were depleted by pig lung perfusion (hDAF(-/-)/AbAbs, hDAF(+/ )/AbAbs) before the experiment. This combined approach was augmented by adding soluble complement receptor 1 (sCR1) to the perfusate in one further group (hDAF(+/-)/AbAbs/sCR1). RESULTS: HDAF(-/-) lungs perfused with unmodified human blood were rejected after 32.5 min (interquartile range, IQR 5 to 210). HDAF(+/-) lungs survived for 90 min (IQR 10 to 161, P = 0.54). Both groups showed a rapid rise in pulmonary vascular resistance (PVR), which is a characteristic feature of hyperacute rejection (HAR). This phenomenon was blunted in the hDAF(+/+) group, although survival (48 min, IQR 14 to 111) was not further prolonged. Antibody depletion (AbAbs) led to a significant increase in survival time (hDAF(-/ )/AbAbs: 315 min, IQR 230 to 427; hDAF(+/-)/AbAbs: 375 min, IQR 154 to 575), reduced PVR and less complement production. Addition of sCR1 reduced complement elaboration but did not further improve survival (200 min, IQR 128 to 580) and surprisingly tended to increase PVR. CONCLUSIONS: Depletion of xenoreactive antibodies is more effective than membrane-bound complement regulation to blunt hyperacute rejection of pulmonary xenografts, but even the combined approach including soluble-phase complement inhibition is not sufficient to reliably prevent organ failure within hours. It therefore seems likely that other factors independent of antibody and complement contribute to HAR in this model. PMID- 15785374 TI - Treatment of streptozotocin-induced diabetes mellitus in rats by transplantation of islet cells from two major histocompatibility complex disparate rats in combination with intra bone marrow injection of allogeneic bone marrow cells. AB - BACKGROUND: We have established a new method for the transplantation of allogeneic pancreatic islets obtained from two different rat strains in combination with a newly developed bone marrow transplantation (BMT) method in which bone marrow cells (BMCs) are directly injected into the bone marrow cavity (intra bone marrow BMT [IBM-BMT]). METHODS: Streptozotocin-induced diabetic Brown Norway (BN: RT1A(n)) rats were injected with fludarabine, irradiated with 5.0 Gy x 2, and BMCs from two allogeneic rat strains, Fischer 344 (F344: RT1A(1)) and PVG (PVG: RT1A(c)), were then directly injected into the bone marrow cavity (IBM BMT). Simultaneously, approximately 600 pancreatic islets (PIs) from F344 and PVG rats were mixed and transplanted into the liver by way of the portal vein. RESULTS: All the recipients thus treated showed normoglycemia 30 days after the treatment. Hematolymphoid cells were completely reconstituted with the two donor type cells, and immunologic tolerance to F344 and PVG major histocompatibility complex (MHC) determinants were induced. CONCLUSIONS: The transplantation of PIs from two MHC-disparate donors was completely achieved in combination with IBM BMT, resulting in the improvement of blood glucose levels and the amelioration of diabetes mellitus. PMID- 15785375 TI - Peripheral nerve allografts stored in green tea polyphenol solution. AB - BACKGROUND: We previously demonstrated the successful 1-month storage of peripheral nerve segments in a green tea polyphenol extract. We investigated whether this method could reduce the donor-host immune reaction associated with peripheral nerve allotransplantation. METHODS: Sciatic nerve segments (20 mm long) were harvested from Dark Agouti (DA) rats, stored in polyphenol solution (1 mg/mL) for 1 month, and transplanted into recipient major histocompatibility complex-mismatched Lewis rats to bridge 15-mm-long sciatic nerve gaps (polyphenol treated allograft group). The controls were an isograft group (nerve segments harvested from Lewis rats were immediately transplanted into Lewis rats), a polyphenol-treated isograft group (nerve segments harvested from Lewis rats were treated by polyphenol in the same method and transplanted into Lewis rats), and a fresh allograft group (nerve segments harvested from DA rats were transplanted into Lewis rats without storage). To investigate the origins of the cells in the transplanted nerves, sciatic nerve segments harvested from the male DA rat donors were transplanted into female Lewis rat recipients; genomic DNA was extracted from each nerve segment and amplified by polymerase chain reaction using primers specific for the rat sex-determining region of the Y-chromosome (Sry). RESULTS: Nerve regeneration in the polyphenol-treated allograft group was similar to that in the isografted group. Sry-specific bands were detected in all samples in the sex-mismatched polyphenol-treated allograft specimens despite their major histocompatibility complex incompatibility. CONCLUSIONS: Storage in green tea polyphenol solution can reduce both ischemic damage to nerve tissue and donor host immune reactions after allotransplantation. PMID- 15785376 TI - Peritransplant immunoadsorption: a strategy enabling transplantation in highly sensitized crossmatch-positive cadaveric kidney allograft recipients. AB - BACKGROUND: Kidney transplant recipients with a current positive complement dependent cytotoxicity crossmatch (CDCXM) are at high risk for hyperacute rejection and graft loss. Immunoadsorption (IA) represents an efficient strategy to remove donor-specific alloantibodies. In this analysis, we evaluated effectiveness of peritransplant IA as an anti-humoral strategy to overcome a current positive CDCXM in presensitized renal allograft recipients. METHODS: Between 1999 and 2003, 40 high risk cadaveric kidney allograft recipients (median CDC panel reactive antibody [PRA] level, 77%; number of retransplants, n = 38) were subjected to peritransplant IA with protein A (one pretransplant IA session followed by a course of repeat posttransplant IA sessions) in addition to preemptive antilymphocyte antibody therapy. RESULTS: In nine of these patients, a current positive CDCXM was rendered negative by a single pretransplant IA session. Thirty-one recipients had a negative CDCXM already before pretransplant IA. No difference in graft survival was found between CDCXM-positive and CDCXM negative recipients (3-year graft survival, 78% vs. 71%, P = 0.6). Comparable rates of immunological graft loss at 3 years were observed (11% vs. 13%, P = 0.8). Patient groups did not significantly differ with respect to median serum creatinine at 1 year (1.23 mg/dL [CDCXM-positive] vs. 1.57 mg/dl [CDCXM negative], P = 0.07) and at the end of follow-up (median 32 months; 1.19 mg/dL vs. 1.63 mg/dL, P = 0.06). Moreover, patient groups showed similar rates of biopsy-proven cellular rejection (11% vs. 20%) or C4d-positive graft dysfunction (33% vs. 32%). CONCLUSION: Our results demonstrate that peritransplant IA enables successful cadaveric kidney transplantation in the context of a positive CDCXM. PMID- 15785377 TI - Infectious enteritis after intestinal transplantation: incidence, timing, and outcome. AB - BACKGROUND: The study reviews the incidence, timing, and outcome of infectious enteritis (IE) after intestinal transplantation (ITx). METHODS: A retrospective review of all patients who underwent ITx at a single institution between 1991 and 2003 was undertaken using database and medical records. Standard statistical analyses were performed. RESULTS: Of 33 ITx recipients, 13 (39%) developed 20 culture- or biopsy-proven episodes of IE. Recipient demographics included the following: 10 males, median age 34 (10-585) months, 11 liver + intestine grafts, and two isolated intestine grafts. Infections were diagnosed a median of 76 days (32-1,800 days) after ITx. There were 14 viral (one cytomegalovirus, eight rotavirus, four adenovirus, one Epstein-Barr virus), three bacterial (Clostridium difficile), and three protozoal (one Giardia lamblia, two Cryptosporidium) infections. The bacterial infections tended to present earlier than the viral infections, and the most frequent presenting symptom was diarrhea. Complete resolution was achieved in 17 (94%) incidences with the appropriate antimicrobial or conservative therapy. It was interesting that there were seven rejection episodes documented by biopsy at the approximate time of diagnosis of IE. There were two graft losses: one because of adenoviral enteritis and one because of rejection after rotavirus enteritis. Three-year patient survival is 74% with no deaths directly attributable to IE. CONCLUSIONS: IE can occur in 39% of recipients after ITx. Viral agents are the cause in two thirds of the cases. With supportive care and appropriate treatment, resolution is possible in the majority of cases. Differentiating rejection and infection on histopathology can be difficult and relies on cultures and immunostaining. PMID- 15785378 TI - Effects of candesartan on glomerular hemodynamics and permselectivity in patients with favorable renal allograft function. AB - BACKGROUND: Distinct effects of angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers on glomerular perfusion and permselectivity are important determinants of the substances nephroprotective quality. In renal allograft recipients, however, specific effects of angiotensin antagonism on glomerular function have not been evaluated so far. METHODS: Twenty patients with favorable allograft function were included into a prospective study within the first year after renal transplantation. Glomerular filtration rate, renal plasma flow, albuminuria, and the fractional clearances of neutral dextrans were determined at baseline and after 3 months of treatment with candesartan. Ten individuals after renal donation served as controls for the baseline evaluation. RESULTS: Compared with the control group, the allograft recipients had a higher renal-vascular resistance and a lower glomerular filtration rate. Albuminuria was significantly higher; however, the difference in the dextran sieving curve was not statistically significant. Apart from mild changes in biochemical parameters, the therapy with candesartan led to a rise in serum creatinine along with a nonsignificant drop in the glomerular filtration rate. There was a highly significant drop in filtration fraction and albuminuria. Glomerular permselectivity clearly improved for a range of dextran molecular diameters from 43 Angstrom up to 73 Angstrom. CONCLUSION: A therapy with candesartan has distinct effects on glomerular function in patients after renal transplantation. A drop in filtration fraction along with an improvement in glomerular permselectivity and albuminuria point to a nephroprotective quality that should lead to a systematic clinical evaluation of candesartan even in patients with favorable renal allograft function. PMID- 15785379 TI - Induction therapy with basiliximab versus Thymoglobulin in African-American kidney transplant recipients. AB - BACKGROUND: It has been suggested that the use of antilymphocyte induction therapy in African-American (AA) renal transplant recipients reduces the risk of acute rejection (AR) and improves graft survival. It is not clear whether the efficacy of basiliximab (BSX) is different from that of Thymoglobulin (ATG) in this regard. METHODS: We retrospectively assessed the effect of induction therapy with BSX versus ATG in 88 AA renal allograft recipients receiving transplants at our center between July 2001 and June 2003 and followed for 19+/-7 months. All patients were maintained on mycophenolate mofetil, prednisone, and either tacrolimus or sirolimus. Study endpoints included patient and graft survival, graft function, and incidence of AR and cytomegalovirus infection. Regression models were used to evaluate the independent effect of each induction agent on these endpoints. RESULTS: Thirty-six patients received ATG, and 52 received BSX. The groups were comparable with regard to donor race and age, and recipient sex, body mass index, human leukocyte antigen (HLA) matching, and hepatitis C virus serostatus. The ATG group was younger, more likely to receive retransplant, had longer duration of end-stage renal disease and higher panel reactive antibody, and was less likely to receive live-donor organs. However, after adjusting for all these variables, graft outcomes, as well as renal function, were comparable between the two induction groups. We found that the degree of HLA mismatch, delayed graft function, and AR were the only significant predictors of graft loss. CONCLUSION: The results of our study suggest that the choice of induction agent may not have a major impact on graft outcomes in AA renal-allograft recipients. PMID- 15785381 TI - Leukemia inhibitory factor is linked to regulatory transplantation tolerance. AB - BACKGROUND: The specific regulation of allo-tolerance in vivo occurs within a complex microenvironment and involves co-operation between a small proportion of different cell types within the spleen or draining lymph node. By analyzing unmanipulated whole spleen cell populations we have aimed to mimic this in vivo situation to identify critical signaling molecules in regulatory allo-tolerance. METHODS: We compared the kinetics of cytokine release and induction of signaling proteins in (BALB/c-tolerant)CBA, versus (BALB/c-rejected)CBA, spleen cells after challenge with BALB/c antigen. RESULTS: The distinguishing features of allo tolerance were Foxp3 protein expression, LIF release, and increased levels of STAT3. Comparison of isogenic clones of Tr1, Th1, and Th2 cells revealed that only the regulatory Tr1 cells are characterized by both LIF and IL10 release. CONCLUSIONS: Overall, our findings demonstrate that allo-antigen driven signaling events can be detected within a whole spleen cell population and identify a role for LIF in the regulation of transplantation tolerance in vivo. PMID- 15785380 TI - In vitro depletion of tissue-derived dendritic cells by CMRF-44 antibody and alemtuzumab: implications for the control of Graft-versus-host disease. AB - Graft-versus-host disease (GvHD), a life-threatening complication of bone marrow transplantation, is initiated by donor T cells reacting to recipient dendritic cells (DC). GvHD can be controlled by attenuating donor T cells, but few strategies exist to target DC, particularly resident tissue DC, despite recent evidence of their importance. In this report, CMRF-44, a mouse monoclonal IgM reactive to human DC, is tested against human Langerhans cells (LC) in vitro. CMRF-44 antigen is expressed at low level on fresh LC but is up-regulated 40-60 fold during migration. CMRF-44 and complement kill more than 97% of migratory LC in vitro and inhibit allostimulation by LC up to 95%. In comparison, alemtuzumab, which binds CD52, reacts weakly with primary LC and fails to induce significant lysis with complement (less than 5%). These results highlight the potential of new therapeutic antibodies active against tissue DC to control graft-versus-host reactions. PMID- 15785382 TI - Cyclosporine A does not alter ultrasonic indices of renal blood flow: a potential tool for differentiating toxicity from acute rejection? AB - The narrow therapeutic window of cyclosporine A (CsA) means its use is controlled by pharmacokinetic monitoring. However, pharmacokinetics do not always reflect the functional effects of a drug--its pharmacodynamics, such as vasoconstriction. We developed a technique for measuring renal blood flow and used a pig model to determine whether CsA-induced renal vasoconstriction could be detected, thus offering a tool for pharmacodynamic therapeutic drug monitoring. This has been shown to differentiate acute rejection from acute tubular necrosis. Power Doppler intensitometry was used to assess relative vascular volume, and the renal arteriovenous transit time was determined with an intravenous microbubble bolus. Measurements were taken before and at intervals after an intravenous bolus of CsA (10 mg/kg). There was no correlation between index and CsA concentration. Lack of detectable effect after CsA administration to high concentrations suggests that this technique may be able to differentiate CsA toxicity from acute rejection. PMID- 15785383 TI - Whole organ pancreas recovery using ultrasonically activated shears. PMID- 15785384 TI - Mycophenolate mofetil is compatible with CD28/CD154 costimulatory blockade in preventing transplant rejection. PMID- 15785385 TI - Drug interaction between itraconazole and sirolimus in a primary renal allograft recipient. PMID- 15785386 TI - Hepatitis C virus recurrence and idiopathic thrombocytopenic purpura after liver transplantation in adult patients: role of splenectomy. PMID- 15785387 TI - Pure red cell aplasia associated with parvovirus B19 infection in renal transplantation: the first case report in Mexico. PMID- 15785388 TI - Endoscopic stenting as first-line treatment in duct of Lushka leaks after choledochocystic anastomosis in two cases of liver transplantation. PMID- 15785389 TI - Recombinant factor VIIa for refractory hemorrhage after lung transplantation. PMID- 15785390 TI - Intravenous immunoglobulin and thymoglobulin induction treatment in immunologically high-risk kidney transplant recipients. PMID- 15785391 TI - Stump transureteroureterostomy in en bloc kidney transplantation. PMID- 15785392 TI - Consideration of preanalytical conditions to use circulating matrix metalloproteinases as diagnostic markers. PMID- 15785393 TI - Preanalytical conditions and circulating matrix metalloproteinases. PMID- 15785394 TI - Probable interaction of bergamottin and cyclosporine in a lung transplant recipient. PMID- 15785395 TI - Sun drop citrus soda and cyclosporine interaction: comments on causality and recommendations. PMID- 15785396 TI - [Pediatric pathology]. PMID- 15785397 TI - [Hirschsprung's disease: practical considerations]. AB - Hirschsprung's disease (1/5000 live births) is defined by the congenital absence of neuronal cells in the nervous plexuses in the distal part of the digestive tract. The disease affects the rectum and sigmoid colon in 80% of cases, or is more extensive. Hirschsprung's disease is suspected in cases of low gastrointestinal obstruction in the neonatal period, or in cases of chronic severe constipation in childhood. It is diagnosed by pathological examination of rectal biopsies that include the submucosa. After standard staining, multiple sections are scrutinized for neuronal cells. Acetylcholinesterase staining is performed on a frozen fragment to demonstrate the hyperplasia of cholinergic fibers that is very suggestive of Hirschsprung's disease. This hyperplasia decreases from the rectum to the splenic flexure of the colon. Hyperplasia of extrinsic nerve fibers and rarefaction of neuromuscular junctions in Hirschsprung's disease may be demonstrated immunohistochemically. Differential diagnosis includes chronic intestinal pseudo-obstructions. The treatment for Hirschsprung's disease is, most often, anastomosis of the normally innervated gut to the anal canal. Peri- or pre-operative biopsies assist surgery, but their interpretation is difficult in the transitional zone. The examination of the surgical specimen allows measurement of the aganglionic segment and transitional zone. Different genes (RET, most often) may be involved in sporadic or familial Hirschsprung's disease. Hirschsprung's disease is associated with other digestive or extra-digestive abnormalities in 5 to 30% of patients. Associated abnormalities may delay the diagnosis and treatment of Hirschsprung's disease. PMID- 15785398 TI - [Hermaphroditism pathology]. AB - Hermaphroditism is a general term referring to all discrepancies between phenotype and genotype of sex development. It must be preferred to sexual ambiguity which refers mainly to external genitalia anomalies. Hermaphroditism is studied on an historical and pathogenetical perspective. Short embryological summaries are integrated. The defects of sexual differentiation due to hormone deficiency are first studied: androgen insensitivity, steroid 5 alpha-reductase 2 deficiency, defects of testosterone synthesis, persistant mullerian ducts syndrome. Sexual determinism deficiencies are then presented: Turner syndrome, XX males, pure gonadal dysgenesis, true hermaphroditism, mixed gonadal dysgenesis, Drash and Frasier syndrome. Tumors of dysgenetic gonads followed. Mixed tumors developed in dysgenetic gonads are gonadoblastoma and dysgerminoma. Sex cord tumors are androgen insensitivity associated tumors, Leydig cells tumors and adrenal cell inclusion tumors. Sex reversion genes open new perspectives. PMID- 15785399 TI - [Nephrogenic rests and nephroblastomatosis]. AB - The term nephrogenic rests has been proposed for abnormally persistant foci of embryonal cells within developmentally normal kidneys. The term nephroblastomatosis is defined as the presence of diffuse or multiple nephrogenic rests. Nephrogenic rests represent precursors of nephroblatoma and they are associated with certain syndromes carrying a high risk for nephroblastoma. In his classification, Beckwith describes different types of nephrogenic rests according to topographic and morphological features. The nephrogenic rests may undergo regression or proliferation with a high risk for development of a nephroblatoma. PMID- 15785400 TI - [Kidney tumors in childhood]. AB - Types of renal tumors arising in children are different from those occurring in adults. Nephroblastoma is the most common (85%) with a clinical outcome which has dramatically improved in the last 30 years. Current classifications are aimed at better adaptation of treatment to each individual case, reducing iatrogenic complications without impairing total cure. Amongst entities which have been recently described or are better known we can find juvenile carcinoma associated with Xp11.2 translocation, renal medullary carcinoma, metanephric tumors, etc. Role of molecular cytogenetics is increasing for classification (and treatment) and this should always be kept in mind when dealing a fresh specimen of childhood renal tumor. PMID- 15785401 TI - [Hepatic tumors in childhood: experience on 245 tumors and review of literature]. AB - This review on the pathology of hepatic tumors in childhood, from a personal series of 245 tumors, focuses on incidence, management, description of frequent tumors such as hepatoblastoma, fibrolamellar carcinoma, and undifferentiated sarcoma for malignant tumors, focal nodular hyperplasia, hepatocellular adenoma, and mesenchymal hamartoma for benign tumors. Malignant and benign entities of recent description, including the following: crowded, small cell undifferentiated and cholangioblastic variants of hepatoblastomas, mesenchymal hamartoma miming hepatoblastoma, liver adenoma and adenomatosis in diabete MODY3 families, gastrointestinal stromal tumor with liver metastasis associated to Carney triad, macronodules in non-cirrhotic portal fibrosis are reviewed. For each entity, the clinical presentation, the diagnostic criteria and the differential diagnosis are described. The role of immunohistochemistry and molecular biology in the diagnosis and identification of new molecular mechanisms triggered by oncogenic activation with new prognostic markers, and therapeutic targets is emphasized. PMID- 15785402 TI - [Peripheral neuroblastic tumors: anatomo pathological classification]. AB - Peripheral Neuroblastic Tumors are classified according to the recommendations of the INPC into four categories and their subtypes: 1/Neuroblastoma stroma poor, undifferentiated, poorly differentiated, and differentiating, 2/ganglioneuroblastoma stroma composite, nodular, 3/ganglioneuroblastoma stroma composite, intermixed and 4/ganglioneuromas stroma dominant, maturing and mature. The classification is based on age and morphologic features of PNT, including the differentiation grade of the neuroblasts and the mitosis-karyorrhexis index. Histopathological classification has prognostic impact in predicting overall and event-free survival allowing the categorisation of PNT into two groups: favorable subset and unfavorable subset. PMID- 15785403 TI - [Cytopathology of malignant blastematous tumors]. AB - Since almost all childhood blastematous tumors belong to the group of non hematological small round cell tumors we describe here the cytopathological aspects of the most frequent of these neoplasms i.e. neuroblastoma, nephroblastoma, rhabdomyosarcoma, peripheral neuroectodermic tumors, and emphasize the role of molecular techniques applicable to cytological material obtained from fine-needle aspiration to help the pediatrician in tailoring therapy. PMID- 15785404 TI - [Pediatric non-Hodgkin lymphomas]. AB - Non Hodgkin lymphoma represent about 5% of childhood cancer. They are the third most common group of cancers in children and adolescents, after cerebral tumours and leukemias. They differ from adult lymphomas as they are all high grade tumours, always diffuse and often associated with extra-nodal diseases. Three main histological subtypes predominate: Burkitt, lymphoblastic and anaplastic large cell lymphomas. As the clinical progression is growing fast and treatment different between the 3 subtypes, the diagnosis must be rapidly done and with accuracy. PMID- 15785405 TI - [Melanocytic nevi in children]. AB - Melanocytic nevi in children can be acquired or congenital. The vast majority are benign but diagnosis can be difficult. Nevi in newborns and nodular proliferations in giant congenital nevi can display histological features that lend to their misdiagnosis as melanoma. Acquired melanocytic nevi in children are often Spitz nevi or related nevi. These lesions are too often misdiagnosed as melanoma. On the other hand, clinicians and pathologists must be aware that melanoma does occur in childhood, albeit very rarely. It is exceptionally rare in prepubescent individuals. It can be associated with the following risk factors: giant congenital nevus, family history of melanoma, xeroderma pigmentosum, and immunosuppression. Melanoma in children younger than 10 years of age is extremely rare and has different clinical and histopathological features than those that arise in postpubescents, often being confused with a Spitz's nevus. Consequently, the diagnosis is often made too late. It should be emphasized that thickness is the main prognostic parameter in childhood melanoma and that early diagnosis is also crucial in this age group. The lesion must therefore be examined in a complete excision and not in a partial biopsy, and if atypical features are present it must be reviewed by an expert. On the other hand extreme caution should be exercised when diagnosing melanoma in children, as some benign lesions exhibit similar histologic features. PMID- 15785406 TI - [Fibroblastic-myofibroblastic proliferations of childhood and adolescents]. AB - Fibroblastic-myofibroblastic proliferations of childhood and adolescents form a clinical and morphologic spectrum from benign reactive processes and pseudosarcomas, to fibromatoses, to various types of sarcoma. The diagnosis is challenging because of clinical and morphologic similarities, lack of specific immunohistochemical markers for different types of fibroblastic-myofibroblastic tumors, and limited molecular genetic information. Careful attention to clinical, macroscopic, and histopathologic features permits classification in most cases. This review focuses on the pathologic features of fibroblastic-myofibroblastic tumors with a predilection for children and adolescents. PMID- 15785407 TI - [Lynch syndrome pediatric: about a case]. AB - A duodenal carcinoma centered on the vater ampulla is reported in a child of 15 years with Lynch syndrome. This case illustrates the problem of early diagnosis and treatment of Lynch syndrome associated tumors. PMID- 15785408 TI - [CD10 expression in a case of microvillous inclusion disease]. AB - All over the causes of intractable diarrhea of infancy, microvillous inclusion disease is a rare congenital defect of intestinal brush border of unknown aetiology. An autosomal recessive inheritance is suggested by cases occurring in siblings and high incidence of consanguinity. The prognosis of the disease is extremely poor, as life can be sustained only by total parenteral nutrition. Combined bowel-liver or bowel transplantation is regarded as the only potentially life-saving therapy. We report a case of microvillous atrophy who undergone a combined bowel, colonic and liver transplantation, and discuss the tools allowing the light microscopic diagnosis. PMID- 15785409 TI - [Stromal gastrointestinal tumors in children: about a case]. AB - Stromal gastrointestinal tumors or GIST are undifferentiated mesenchymal tumors of gastrointestinal tract that showed characteristic c-kit expression. GIST are very rare in children and only 21 cases are reported in literature. The authors present a GIST arising in the stomach of a 7-year-old girl and revealed by severe anaemia. The patient underwent surgical resection and chemotherapy because of high-risk markers. Positive diagnosis and prognosis of paediatric GIST are discussed with literature data. PMID- 15785411 TI - [A precocious ovarian tumor]. PMID- 15785410 TI - [Benign epithelial cells and Tamm-Horsfall protein in lymph nodes from nephrectomy specimens with nephroblastoma: a diagnostic pitfall]. AB - When a lymph node metastasis is discovered during the pathology examination of surgical specimens with nephroblastoma, the tumour is classified as stage III, according to the classification of the International Society of Paediatric Oncology (SIOP 2001), and post-operative intensive treatment of the patient includes irradiation and chemotherapy. Benign epithelial cells in perinephric lymph nodes in cases of renal tumours in childhood may be confused with metastases. They are often associated with an accumulation of Tamm-Horsfall protein in lymph node sinuses. This case illustrates latero-aortic lymph node complexes of benign epithelial cells and Tamm-Horsfall protein in a 16-month-old girl with surgical resection of a nephroblastoma after pre-operative chemotherapy. The nephroblastoma was predominantly epithelial and was classified as SIOP stage I. There were accumulations of Tamm-Horsfall protein in lymph node sinuses, in lymphatic vessels, in the kidney outside the tumour, and in the renal sinus. This association of epithelial cells with deposits of Tamm-Horsfall protein and their resemblance to cells of the distal convoluted tubules favour a diagnosis of benignity. A definitive diagnosis is supported by the small size of the cells, their bland nuclei, and morphological features differing from those of tumour cells, but deposits of Tamm-Horsfall protein may be associated with true lymph node metastases. PMID- 15785412 TI - [An unusual pediatric tumor]. PMID- 15785413 TI - [Helical CT of abdominal aorta]. AB - The purpose was to illustrate that single or multirow spiral CT angiography has become the main modality for the examination of the abdominal aorta. Our multidisciplinary group investigates weekly 15 abdominal aortas, with a single slice spiral CT since 1995, and a 4 row detector spiral CT since 1999. CT scans protocols and parameters optimization for the 2 CT systems are described. Indications and results of CT to investigate the various abdominal aortic diseases are illustrated and discussed: aneurisms (pretherapeutic staging, selection of indications of endovascular stent-grafting); obstructive aorto arteriopathies (pretherapeutic staging, follow up after endovascular revascularization); dissection (diagnosis, pretherapeutic staging, follow up); inflammatory diseases; normal patterns and principal complications after surgery. Situations where multislice CT is superior to single slice CT and compared advantages of MRI are discussed. CT is at the present time the main technique to image abdominal aorta. Multislice CT is especially interesting to explore the whole aorta, poly traumatisms, and to plan and follow up endovascular treatments. PMID- 15785414 TI - [Understanding CAD (computer-aided diagnosis) in mammography]. AB - Generalization of breast screening programs requires efficient double reading of mammograms, which allows reduction of false negative interpretations, but it may be difficult to achieve. CAD (Computed Aided Detection) systems are dramatically improving and can now assist in the detection of suspicious mammographic lesions, either suspicious microcalcifications, masses or architectural distortion. Characterization of the lesions is improving as well. CAD mammography might complete or substitute to "human" double reading. The aim of this review is to present the main CAD systems commercially available, review the principles of CAD and discuss the results of CAD mammography. Specifically, the role of CAD within breast screening program, according to the results of recent prospective studies will be discussed. PMID- 15785415 TI - [Linitis plastica]. PMID- 15785416 TI - [Long term clinical follow up of vertebral hemangiomas treated by percutaneous vertebroplasty]. AB - PURPOSE: Long term clinical follow up of vertebral hemangiomas (VH) treated by percutaneous vertebroplasty. PATIENTS AND METHODS: A retrospective analysis of 19 patients treated fro 21 VH associated with chronic pain in 16 cases, using pecutaneous vertebrosplasty. One patient underwent both vertebroplasty and surgery. Long term follow up (mean 38.6 months; 16-83 months) consisted in the evaluation of residual or secondary pain, its impact on daily activities (using the Oswestry Low Back Pain scale) and the need for pain medication. RESULTS: Long term follow up found 56.2% of patients to be asymptomatic (9/16) and 31.2% (5/16) with occasional pain without much impact on daily activities and no need for chronic use of pain medication. The treatment was ineffective in 2 cases. Three patients treated for aggressive hemangiomas remained painless. There was no complication due to the treatment. No poor outcome was noted. CONCLUSION: Percutaneous vertebroplasty appears to be an effective technique in the treatment of VH with approximately 90% of satisfactory results in the short or long term period. PMID- 15785417 TI - [Dynamic evaluation of lower limb arteries using the ECTRICKS MRI technique]. AB - PURPOSE: This article presents a new MR angiography technique, Elliptic Centric Time Resolved Imaging on Contrast KineticS (ECTRICKS), which allows dynamic evaluation of the lower limb arteries opacification when venous return prevents adequate diagnostic evaluation of calf arteries. PATIENTS AND METHODS: Comparative examinations could be compared. Additional arteries in 59 patients using a standard MRA technique and the ECTRICKS technique. RESULTS: A total of 33 examinations could be compared. Additional arteries could be evaluated in 10% of our patient population using the ECTRICKS technique, and in 20% for our diabetic population. Arterial evaluation was better with the ECTRICKS technique in patients with diabetes and venous insufficiency. CONCLUSION: ECTRICKS is a good complementary sequence for evaluation the calf arteries in patients with diabetes or early venous return. PMID- 15785418 TI - [Image guided drainage of pelvic fluid collections: results in 42 patients]. AB - PURPOSE: To review the etiology, location, and morphology of pelvic collection as well as the technique and results of image guided drainage. MATERIALS AND METHODS: From June 1996 to August 2002, we performed image guided drainage of pelvic fluid collections in 21 males and 21 females. In patients where a direct contact between the collection and the endocavitary probe was present, the drainage was performed either by transrectal or transvaginal approach using 10F, 12F, 14F or 16F catheters according to the viscosity of the fluid. When the patients were no longer septic, when drainage had stopped, the drains were removed at day 5. When a fistula was present, the drain was left in place until the fistula healed. RESULTS: The most common location of pelvic collections was the cul-desac (43%). A total of 81% of pelvic abscesses were digestive in origin, either from the colon or appendix. Transrectal or transvaginal drainage was possible in 83% of cases. Mean follow-up was 41 months. No drainage related complication was recorded. In two patients with collections of clear fluid, a simple aspiration was performed without insertion of a drain. In the 40 other patients, a drainage catheter was inserted. Twenty-nine patients were cured after 15 days of drainage. Two patients had recurrent collections. Image guided drainage failed in five patients, and all underwent successful surgical management. CONCLUSION: Image guided drainage of pelvic collections is a safe and effective procedure. Failures were due to initially undiagnosed pathology requiring surgical treatment. PMID- 15785419 TI - [MRI of hippocampal malformations in patients with intractable temporal lobe epilepsy]. AB - OBJECTIVE: In some patients with temporal lobe epilepsy, recent MRI studies have revealed several morphological features indicative of discrete hippocampal malformation (HM). Its prevalence is unknown and the relationship between the HM and the origin of seizures has never been investigated. Our purpose is to define the MRI findings of this new entity and to determine its incidence in a group of patients and in a control group in order to evaluate its clinical significance. MATERIALS AND METHODS: MR imaging findings in 97 patients suffering from medically intraceable temporal epilepsy were prospectively evaluated during the preoperative evaluation of surgical candidates. The MR-imaging protocol included oblique coronal slices perpendicular to the temporal lobes using high resolution T2 weighted (HR TSE T2), Fluid attenuated inversion recovery (FLAIR) and inversion-images. This protocol has been completed by axial FLAIR images and axial and sagittal IR images of the whole brain. Coronal HR TSE T2 images were performed in 50 healthy control subjects. Cerebral lesion and hippocampal morphology were evaluated in both groups. RESULTS: Fourteen patients (14%) showed hippocampal morphological modification. The most frequent and specific findings were lack of visualization of the internal hippocampal (lack of linear T2 hypointensity within the hippocampus) and the abnormal shape (pyramidal, vertically oriented or globular-shaped). Other signs were: abnormal position of the hippocampus (medically located hippocampus) and vertical collateral sulcus. Cases without visualization of the internal structure of the hippocampus were considered as a complete form of HM and were correlated with temporal epilepsy. A vertical collateral sulcus was observed in some control group subjects. CONCLUSION: Complete forms of HM could be considered as epileptogenic lesions. Nevertheless, interpretation of the incomplete form is delicate: the abnormal angle of the collateral sulcus can be encountered in healthy subjects and could therefore be considered a normal variant. PMID- 15785421 TI - [Pulmonary hamartochondroma: a rare cause of solitary pulmonary nodule in children]. AB - Solitary pulmonary nodules are rare in children. We report an eleven year-old girl evaluated for recurrent respiratory symptoms diagnosed with an intraparenchymal pulmonary hamartochondroma. Spiral computed tomography showed a pulmonary nodule in the middle lobe, 17 mm in diameter, without any specific features. In asymptomatic adult patients, guidelines for the management of solitary pulmonary nodules have been described. The management of solitary pulmonary nodules in pediatric patients is discussed. PMID- 15785420 TI - [Stent-graft treatment of a ruptured aortic graft: a case report]. AB - The authors report the clinical and imaging features of a patient with rupture of an aortoiliac graft successfully treated by endovascular approach. The endovascular treatment is easy to perform and effective. The main pitfall of this technique is the limited availability of stent-grafts. PMID- 15785422 TI - [Intraosseous meningioma of the skull: radiologic pathologic correlation]. AB - Intraosseous meningiomas are rare ectopic meningiomas. The authors report the case of a hyperostotic intraosseous meningioma of the parietal bone without dural extension. The preoperative imaging findings, as well as imaging features of the surgical specimen and pathologic findings are discussed. PMID- 15785423 TI - [Isolated basilar artery dissection in a case of osteogenesis imperfecta]. AB - Vascular dissection and osteogenesis imperfecta are very rarely associated. The authors report a unique case of basilar artery dissection and discuss the diagnostic imaging work-up. PMID- 15785424 TI - [What is your diagnosis?]. PMID- 15785425 TI - Autosomal recessive and dominant polycystic kidney diseases. AB - It is possible to identify renal cysts in several subjects by ultrasonography imaging techniques. Among the inherited polycystic kidney diseases we include autosomal recessive polycystic kidney disease (ARPKD) and autosomal dominant polycystic diseases such as von Hippel-Lindau disease, tuberous sclerosis complex (TSC1 and TSC2), and autosomal dominant polycystic kidney disease (ADPKD). ARPKD is a rare disease, related to PKHD1 gene, located on chromosome 6p21, that encodes a protein named polyductin/fibrocystin. Pathoanatomical features are bilateral kidney involvement with multiple microcysts, and invariably liver involvement with portal and interlobular fibrosis. A single genetic defect leads to different degrees of renal and hepatic involvement with very different phenotypes and different clinical outcome, in the same family too. ARPKD clinically may show 4 different forms: perinatal, neonatal, infantile, and juvenile. ADPKD is much more frequent (1: 400-1000 live births), and can arise from mutations in 2 different genes, named PKD1 located on chromosome 16p13.3, and PKD2 located on chromosome 4q21-23. The proteins encoded by the PKD1 and PKD2 genes are named polycystins which play crucial roles in several biologic processes. To explain the focal lesions that affected different organs and tissues the "double hit" theory has been proposed (germinal mutation plus somatic mutation on PKD1 or PKD2). Recently, biologic evidence documented the crucial role of the renal primary cilia on the formation of polycystins to induce cystogenesis. ADPKD may be clinically characterized by abdominal pain, hypertension, episodes of gross hematuria, headache, renal stones, aortic and cerebral aneurysms, mitral valve prolapse, and polycystic liver disease. ADPKD is slowly progressive disease responsible for up 10% of end stage renal failure (ESRF) in every country of the world. Male sex, PKD1 gene, episodes of gross hematuria, and the precocity and severity of hypertension play an important role in the progression of renal disease to ESRF. PMID- 15785426 TI - Renal effects of PPARalpha-agonists. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a member of the large nuclear receptor superfamily. Its main role is to activate genes involved in fatty acid oxidation in the liver, heart, kidney and skeletal muscle. While they are mainly used as hypolipidemic agents, PPARalpha agonists may also be postulated to exhibit renoprotective effects. This review summarizes current knowledge regarding the effects of PPARalpha agonists on the kidney. PMID- 15785427 TI - The US color Doppler in acute renal failure. AB - Imaging techniques, especially ultrasonography and Doppler, can give an effective assistance in the differential diagnosis of acute renal failure (ARF). An resistance Index (RI) value >0.75 is reported as optimal in attempting differential diagnosis between acute tubular necrosis (ANT) and prerenal ARF. In hepatorenal syndrome (HRS) RIs is very increased. In some renal vasculitis, as nodose panarteritis (PN), hemolytic-uremic syndrome (HUS), thrombotic thrombocytopenic purpura (TTP), parenchymal perfusion is reduced and RI increased. In lupus nephritis the RI values are correlated with creatinine level and normal RI are considered as a good prognostic tool. In acute primitive or secondary glomerulonephritis (GN), RI value is normal, with diffuse parenchymal hypervascularization. In acute crescentic and proliferative GN and tubulo interstitial disease, color Doppler (CD) and power Doppler (PD) reveal a decreased renal parenchymal perfusion, which correlates with increased RI values. In acute thrombosis of renal artery, US color Doppler (DUS) reveals either an absence of Doppler signal or a tardus-parvus pulse distal to the vascular obstruction. In this situation it is possible to visualize hyperthropic perforating vessels that redirect their flow from the capsular plexus to the renal parenchyma. In acute thrombosis of the renal vein Doppler analysis of parenchymal vessels reveals remarkable RI values, sometimes with reversed diastolic flow. In postrenal ARF an adjunct to the differentiation between obstruction and non obstructive dilatation can be found through RIs. Diagnostic criteria of obstruction as reported by literature are: RI>0.70 in the obstructed kidney and, mostly, a difference in RI between the 2 kidneys >0.06-0.1. PMID- 15785428 TI - Catheter knots in the urinary tract. AB - The use of catheters in the urinary tract, both in children and adults, has not been without complications. Many procedures performed in the urinary tract, including urethral or suprapubic catheterization of the bladder, urethrography, and ureteral catheterization, have showed complications, one of which is the formation of catheter knots. This complication, although very rare, yet it is very annoying. In this study, we searched (in winter 2002, employing Medline) and reviewed published reports of urinary tract catheter knots, and present measures for prophylaxis, diagnosis and proper management of this rare complication. PMID- 15785429 TI - Routine insertion of permanent peritoneal dialysis catheters in the nephrology ward. The sliding percutaneous technique. AB - AIM: A prerequisite to the technical success of peritoneal dialysis in its different modifications is a safe, reliable, correctly positioned, and functioning peritoneal catheter. The ideal method for insertion of permanent catheters (PC) remains debatable. The most commonly used technique is an open surgical approach, by minilaparatomy. Others have proposed a blind approach, using either a trocar method or a modified Seldinger technique or an insertion by means of peritoneoscopy. METHODS: We describe our 5-year experience (May 1997 - June 2002) with 68 percutaneous PC insertions in 63 consecutive patients. A modified technique of percutaneous PC insertion was used that here we call sliding percutaneous technique (SPT): 1) a midline incision is made 4 cm below the umbilicus; the peritoneum is punctured using a straight plastic catheter with a stylet. The catheter is pushed caudally towards the left iliac fossa; 2) a straight stiff 90 cm stylet is inserted through the temporary catheter, which is subsequently removed. The stylet has 2 tips, a blunt proximal one, which is pushed through the catheter into the iliac fossa, and the distal one; 3) having the stylet in situ, a double-cuffed curled-end PC is mounted at the distal tip of the stylet starting from the curl; then, the PC is gently slid down along the stylet as a train does along the rail-way. PC failure was defined as mechanical dysfunction, persistent dialysate leak and persistent peritonitis, or exit site/tunnel infection requiring PC removal. Furthermore, PC failure was defined as early, if occurring in the first 30 days after PC placement, or late, if occurring more than 30 days post-operation. Life-table estimates of PC survival were determined by Kaplan-Meier analysis. RESULTS: Early complications: in the 1st month on dialysis, obstruction to dialysate flow was the commonest cause leading to PC failure. The early actuarial survival (AS) was 90%. Late complications: peritonitis was the commonest cause of PC failure. The late AS, i.e. excluding the catheter failures occurring in the 1st month post-operation, was 82% at 3 years. Global AS, i.e. including both early and late PC failures, was excellent (74% at 3 years). CONCLUSIONS: Even though this study is retrospective and does not compare percutaneous with surgical PC placement techniques, it allows us to state that percutaneous PC insertion is a well tolerated, rapidly performed, side-room procedure that gives excellent results, above all when using SPT. PMID- 15785430 TI - The influence of extracellular fluid volume on serum proteins in peritoneal dialysis patients. AB - AIM: Patients on peritoneal dialysis (PD) often present a deficit of serum protein and especially of albumin, due to its loss through the peritoneum and some others factors. Serum albumin is a marker of survival, of nutritional status and is changed in inflammation in renal failure. Correction of the albumin level is difficult and needs a dietary or other supplementation. Some suspicions exist that a reduction exist that a reduction in overhydration in PD patients may improve serum proteins, particularly albumins. The study tries to prove or to deny the thesis, investigating 27 PD patients (cycler PD, CPD) during 6 weeks. METHODS: Body weight, blood pressure, total serum protein, albumin, C-reactive protein, 24-h urine and volume of the dialysate solution were measured at the first and the final week. The extracellular fluid volume was evaluated by multiple-frequency bioelectrical impedance. Dialysis prescription was changed between the 0- and 6-week to increase the daily ultrafiltrate volume (1.00+/-0.87 1 to 1.32+/-0.85 1, p<0.05). RESULTS: There was a significant reduction in body weight (p<0.001), extracellular fluid volume (p<0.001), systolic blood pressure (p<0.001), diastolic blood pressure (p<0.01). Serum albumin increased significantly (p<0.01), there was a trend to augmentation of total protein and C reactive protein did not change. CONCLUSIONS: The study suggests that overhydration has only a light influence on total serum protein, but impacts significantly the serum albumin level and blood pressure as well. The reduction in body weight can be a helpful way to overcome the hypoalbuminemia and hypertension i.e. to improve the survival time of patients on continuous ambulatory peritoneal dialysis. PMID- 15785431 TI - The utility of fetal urine biochemistry in the management of lower urinary tract obstructions. A report of 3 cases. AB - The aim of this study is to implement the use of biochemical parameters in the analysis of fetal urine to assist with decision making for surgical intervention for fetal obstructive uropathy. Three patients were selected over a 6 month period following ultrasound diagnosis of megabladder and oligohydramnios. Amniocentesis and cystocentesis were performed for karyotyping and biochemical urinary electrolyte evaluation, respectively. Fetal urine biochemistry assisted in decision making with regard to patient and fetal well being. In each of the cases the parameter cutoffs were diagnostic of fetal condition. While the long term benefits of fetal intervention for obstructive uropathy are still debated, we feel that the use of urine biochemical criterIa provide the best current method to select the subjects to be submitted to vesicoamniotic shunt centres. PMID- 15785432 TI - Prevalence and pathogenesis of osteoporosis in patients with inflammatory bowel disease. AB - Decreased bone mineral density is a frequent finding in patients with inflammatory bowel disease. Factors contributing to this are: 1) malabsorption of vitamin D, calcium and possibly vitamin K and other nutrients, 2) treatment with corticosteroids, 3) inflammatory cytokines in inflammatory bowel disease, and 4) hypogonadism induced by the inflammatory bowel disease. Among patients with Crohn's disease from 32% to 38% have osteopenia (Z-scores <-1), and among patients with ulcerative colitis 23% to 25% have osteopenia. The mean deficit was 0.44+/-0.08 Z-scores in the spine in Crohn's disease and 0.34+/-0.08 in ulcerative colitis. A similar deficit was seen in the hip in both conditions. From these deficits, an increase in overall fracture risk of 1.1-1.3 should be expected. The observed excess fracture risk was limited compared to the general population in both Crohn's disease (RR=1.2, 95% CI: 0.9-1.6 for any fracture and 2.2, 95% CI: 1.2-4.0 for spine fractures) and ulcerative colitis (RR=1.1, 95% CI: 1-1.2 for any fracture, and 1.5, 95% CI: 0.9-2.5 for spine fractures). The observed excess fracture risk was close to that expected from the changes in BMD. Despite the limited excess fracture risk, a relatively large percentage of all fractures may be attributable to corticosteroid use among users of corticosteroids. PMID- 15785433 TI - Diagnosis and management of osteoporosis in inflammatory bowel disease. AB - Osteoporosis is a frequent finding in patients with Crohn's disease and ulcerative colitis. The prevalence of vertebral fractures in those patients with significantly reduced bone mineral density is up to 22%. Factors contributing to osteoporosis in inflammatory bowel disease (IBD) patients are treatment with glucocorticoids, increased cytokine production by the inflammation itself, malabsorption and possibly hypogonadism. Therefore, consequent treatment of the underlying IBD and minimising therapy with systemic glucocorticoids, as well as the adequate intake of calcium and vitamin D, may be very important measures to prevent bone loss in IBD. In patients with osteoporosis associated with Crohn's disease or ulcerative colitis, various treatment strategies, such as sodium fluoride and aminobisphosphonates, are discussed. Unfortunately, interventional studies in secondary osteoporosis are often limited by the small study population. The efficacy in prevention of vertebral fractures is not proven in any of the described treatment modalities in these patients. Therefore, guidelines are based on data using bone density as the most accepted surrogate marker and treatment guidelines are based on data from patients with postmenopausal and steroid-induced osteoporosis. PMID- 15785434 TI - Bad to the bone: the effects of liver diseases on bone. AB - Metabolic bone disease (MBD) is a relevant complication of long-standing liver disease. It has been described in association with most types of chronic liver disease both cholestatic and non-cholestatic. It can significantly affect morbidity, and quality of life of these patients. Fractures are also associated with an excess mortality. Recently, the issue of MBD in liver disease has been considered to be such an hot topic to be the subject of 2 recent review/guideline articles produced by the British Society of Gastroenterology and the American Gastroenterological Association. Aim of this paper is to summarize some practical issues regarding this topic and to provide a rapid overview of the main pathophysiological and pharmacological aspects. PMID- 15785435 TI - Postmenopausal osteoporosis and hormone replacement therapy. AB - Osteoporosis is a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. Postmenopausal osteoporosis is a major public health problem. Estrogen deficiency is a key factor in the pathogenesis of postmenopausal osteoporosis. Among the several types therapeutic intervention in osteoporosis, hormone replacement therapy (HRT) has traditionally been seen as the gold standard method of preventing osteoporotic fractures among postmenopausal women.The estrogenic effect on bone is dose-dependent. For oral therapy, studies have demonstrated that doses of 0.625 mg of conjugated equine estrogen and 2 mg of micronized estradiol prevent postmenopausal bone loss. Percutaneous 17 beta-estradiol prevents skeletal bone loss as effectively as oral HRT. Although the greatest benefits from HRT in terms of bone sparing effects can be obtained shortly after the menopause, the literature contains clear evidence that HRT prevents bone loss in all stages of postmenopausal life. However, estrogen therapy must be long-term, possibly lifelong, to have any lasting impact on bone health. One strategy to improve long term continuation of HRT is to reduce the dosage of estrogen and the consequent side-effects of the higher dose HRT. Various studies have assessed the efficacy of low-dose HRT (LD-HRT) as well as the standard dose HRT in the prevention of osteoporosis in postmenopausal women. LD-HRT may enhance patient acceptability and continuation, ensuring adequate bone protection and menopausal symptoms control, and given physicians the possibility to personalize the doses on the basis of each individual patient's needs. PMID- 15785436 TI - Statins and osteoporosis: myth or reality? AB - Several beneficial effects produced by statins over and above the reduction in plasma cholesterol levels, the so-called ''pleiotropic effects'' of statins, have been described. Recent clinical and experimental data have suggested a potential new effect of these drugs, namely a reduction in the risk of osteoporotic fractures. In 1999 the role of statins in bone formation was shown and, after that, observations of large groups of patients have pointed to a reduction in the risk of osteoporotic fractures with the use of statins compared to those using other lipid-lowering drugs or to the control group. The first prospective studies have produced contrasting results as to the effects of therapy with several statins (atorvastatin, fluvastatin, simvastatin) at different doses on biochemical markers of bone remodelling. To date only one randomised trial has been published. This compares the effects of treatment with simvastatin and atorvastatin on the levels of biochemical markers of bone remodelling, but still with non-univocal results: only therapy with simvastatin (but not atorvastatin) reduced the levels of bone-specific alkaline phosphatase. To conclude, observational studies have shown a reduction in the risk of osteoporotic fractures with the use of statins, but it is not yet known whether using these drugs may have a beneficial effect on bone turnover. We must therefore await larger prospective randomised clinical trials before prescribing these drugs in osteoporotic patients. PMID- 15785437 TI - Long term follow up of bone mineral density in patients with primary biliary cirrhosis. AB - AIM: Osteodystrophy is a well known complication of cholestatic liver diseases such as primary biliary cirrhosis (PBC), although the factors predisposing to the development of metabolic bone disease and its natural history have not been well studied. METHODS: We followed a cohort of patients, predominantly post-menopausal females, for changes in bone mineral density over a period of up to 8 years. RESULTS: We found that most patients with PBC have relatively stable lumbar bone density over time, even in the face of progressive liver disease. Only 2 of 22 patients experienced fractures. Both of these patients had low initial bone density measurements. CONCLUSIONS: We recommend that all patients with newly diagnosed PBC undergo bone mineral density measurement so as to identify those at greatest risk of pathological fractures. PMID- 15785438 TI - Prevalence of HFE and TFR2 gene mutation in 118 Ligurian rheumatic patients. AB - AIM: HFE gene is associated to haemochromatosis, an inherited autosomal recessive disorder responsible of an overload of iron in intestine, liver, pancreas, heart, cutis and joints. Articular and periarticular calcifications may occur. H63D mutation may play a role in the pathogenesis of rheumatoid arthritis. METHODS: DNA of 118 consecutive patients (28 males, 90 females, mean age 58.5+/-13.44) living in Liguria and affected by different rheumatic diseases was examined to evaluate the presence of HFE mutations. RESULTS: Analysis data showed that in 45% (53/118) of patients almost one mutation of HFE gene was present and the presence of H63D mutation in the rheumatic patients was particularly elevated. Data obtained in this study have permitted to reveal that 25 patients of 53 (47.1%) with 1 of 11 HFE mutations suffered from symptomatic or silent chondrocalcinosis. CONCLUSIONS: The conclusion is drawn that this mutation may be correlated to various rheumatic diseases. PMID- 15785439 TI - The clinical role of extracellular bioimpedance tomography (Gastro-Mida(x)) in the diagnosis of colorectal diseases. AB - AIM: Gastro-Mida(x) is a non-invasive diagnostic medical device which carries out an extracellular bioimpedance tomographic analysis of the colorectal region with a neural technique interpretation using artificial neural networks to diagnose colorectal diseases. The aim of this spontaneous study carried out in various centres in Italy was to train Gastro-Mida(x) to distinguish normal patients from those with colorectal diseases. METHODS: Patients who were referred to the Endoscopy Units of the 20 centres involved in the study to undergo colonoscopy from September 2002 to December 2003 were included in the study. At least 1 day before colonoscopy, patients underwent Gastro-Mida(x). The training of the bioimpedance tomography in this study considered patients negative (normal) when the colorectal endoscopy was normal or revealed only the presence of haemorrhoids. Patients were considered positive in the presence of diverticula (diverticulosis or diverticulitis), polyps, cancer. For positive patients with more than one pathology, neoplastic or preneoplastic lesions were considered more important than the others and the final diagnosis was made in accordance with this rule. RESULTS: Overall, the 20 centres enrolled 947 patients (males/females: 477/470; average age+/-SD: 57.44+/-13.85 years). The specificity registered after training was 80% (95% CI: 76.5-83.1) and sensitivity was 83.89% (95% CI: 79.5 87.4). The sensitivity of bioimpedance tomography in the subgroup of patients with cancer (N=68) was 88.2% (95% CI: 78.4-93.9). As to polyps, sensitivity was calculated as a whole and according to size classification. The sensitivity of bioimpedance tomography in diagnosing patients with polyps was therefore 78.3% (95% CI: 71.9-83.4). However, sensitivity increased to 83.7% (95% CI: 74.1-90.2) and 83% (95% CI: 70.7-90.8) when the bioimpedance tomography's ability to identify patients with at least one polyp > or = 6 mm and > or = 10 mm in size was taken into consideration. CONCLUSIONS: Gastro-Mida(x) has proven to be a simple, reliable and accurate instrument, once training is completed, in the most common colorectal diseases. The device can therefore be proposed for both diagnosis and screening of colorectal diseases. PMID- 15785440 TI - Colorectal carcinoma that afflicted King Jehoram. AB - This research uses the tools of modern medical science to examine the ancient descriptions of the symptoms suffered by King Jehoram who was affected by some disease. The Biblical texts were examined, and passages relating to the disease that afflicted King Jehoram, who ruled in Jerusalem 843-851 B.C., were closely studied. We have not included any commentaries, but referred only to the words of the Bible exactly as written. The Passages ''...the Lord smote him in his bowels with an incurable disease in the process of time, after the end of two years, his bowels fell out by reason of his sickness: so he died of sore diseases'' indicate that the King suffered from some kind of disease which affected his bowels. Among the various diseases which may be associated with prolapse of the bowel, colorectal carcinoma is the most acceptable. It seems that the colorectal carcinoma was poorly differentiated, invaded perirectal adipose tissue, blood vessels, and/or lymphatic vessels, and/or perineural areas, was lymph node positive and reached the 4th stage with the spread of metastases to the distal organs. Viewed by a modern physician, the story of King Jehoram unfolds as possibly the earliest description of a patient afflicted by colorectal carcinoma. PMID- 15785447 TI - Abstracts of the 7th International Conference of Nuclear Cardiology, 8-11 May 2005, Lisbon, Portugal. PMID- 15785734 TI - The psychological impact of human papillomavirus testing in women with borderline or mildly dyskaryotic cervical smear test results: 6-month follow-up. AB - State anxiety (S-STAI-6), distress (GHQ-12), concern and quality of life (EuroQoL EQ-5D) 6 months after human papillomavirus (HPV) testing in women with borderline or mildly dyskaryotic smear test results were assessed based on a prospective questionnaire study, with 6-month follow-up after the smear test result. Two centres participated in an English pilot study of HPV testing. Participants included two groups of women receiving abnormal smear test results: (tested for HPV and found to be (a) HPV positive (n=369) or (b) HPV negative (n=252)) and two groups not tested for HPV (those receiving (c) abnormal smear test results (n=102) or (d) normal smear test results (n=288)). There were no differences in anxiety, distress or health-related quality of life between the four study groups at 6 months. Levels of concern about the smear test result remained elevated in all groups receiving an abnormal smear test result, and were highest in the group untested for HPV. Predictors of concern across all groups receiving an abnormal smear test were perceived risk of developing cancer, being HPV positive or untested for HPV, sexual health worries and the smear being a woman's first smear test. The raised anxiety and distress observed in women immediately after being informed of an abnormal smear test result and that they are HPV positive was no longer evident at 6 months. Concern about the smear test result was however still raised in these women and those who tested negative for HPV, and particularly among those who did not undergo HPV testing. PMID- 15785735 TI - MET signalling in primary colon epithelial cells leads to increased transformation irrespective of aberrant Wnt signalling. AB - It has been shown that in hereditary and most sporadic colon tumours, components of the Wnt pathway are mutated. The Wnt target MET has been implicated in the development of colon cancer. Here, we show that overexpression of wild-type or a constitutively activated form of MET in colon epithelial cells leads to increased transformation irrespective of Wnt signalling. Fetal human colon epithelial cells without aberrant Wnt signalling were transfected with wild-type or mutated MET constructs. Expression of these constructs leads to increased phosphorylation of MET and its downstream targets PKB and MAPK. Upon stimulation with HGF, the expression of E-cadherin is downregulated in wild-type MET-transfected cells, whereas cells expressing mutated MET show low E-cadherin levels independent of stimulation with ligand. This implies a higher migratory propensity of these cells. Furthermore, fetal human colon epithelial cells expressing the mutated form of MET have colony-forming capacity in soft agar, while cells expressing wild-type MET show an intermediate phenotype. Subcutaneous injection of mutated MET-transfected cells in nude mice leads to the formation of tumours within 12 days in all mice injected. At this time point, mock-transfected cells do not form tumours, while wild-type MET-transfected cells form subcutaneous tumours in one out of five mice. We thus show that MET signalling can lead to increased transformation of colon epithelial cells independent of Wnt signalling and in this way could play an essential role in the onset and progression of colorectal cancer. PMID- 15785736 TI - Targeting of Rad51-dependent homologous recombination: implications for the radiation sensitivity of human lung cancer cell lines. AB - The aim of the present work was to study the role of Rad51-dependent homologous recombination in the radiation response of non-small-cell lung cancer (NSCLC) cell lines. A dose- and time-dependent increase in the formation of Rad51 and gamma-H2AX foci with a maximum at about 4 and 1 h after irradiation, followed by a decrease, has been found. The relative fraction of cells with persisting Rad51 foci was 20-30% in radioresistant and 60-80% in radiosensitive cell lines. In comparison, a higher fraction of residual Dsb was evident in cell lines with nonfunctional p53. Transfection with As-Rad51 significantly downregulates radiation-induced formation of Rad51 foci and increases apoptosis, but did not influence the rejoining of DNA double-strand breaks. Interestingly, wortmannin, a well-known inhibitor of nonhomologous end-joining, also inhibits Rad51 foci formation. In general, there was no correlation between the clonogenic survival at 2 Gy and the percentage of initial Rad51 or gamma-H2AX foci after ionising radiation (IR). The most reliable predictive factor for radiosensitivity of NSCLC cell lines was the relative fraction of Rad51 foci remaining at 24 h after IR. Although most of the Rad51 foci are co-localised with gamma-H2AX foci, no correlation of the relative fraction of persisting gamma-H2AX foci and SF2 is evident. PMID- 15785737 TI - Efficacy of epidermal growth factor receptor-targeted molecular therapy in anaplastic thyroid cancer cell lines. AB - Anaplastic thyroid cancer is one of the most aggressive human malignancies and the outcomes of conventional therapy have been far from satisfactory. Recently, epidermal growth factor receptor (EGFR)-targeted therapy has been introduced as an alternative therapeutic strategy for highly malignant cancers. This study was undertaken to investigate the expression of EGFR in anaplastic thyroid cancer cell lines, and to explore the potential of therapies targeting EGFR as a new therapeutic approach. EGFR was universally expressed in anaplastic cancer cell lines at a variety of levels. Specific EGFR stimulation with epidermal growth factor showed significant phosphorylation of ERK1/2 and Akt, and resulted in marked growth stimulation in an anaplastic thyroid cancer cell line, which highly expressed EGFR. This EGFR-transmitted proliferation effect of the cancer cell line was completely inhibited by gefitinib, an EGFR tyrosine kinase inhibitor. Moreover, growth of xenografts inoculated in mice was inhibited in a dose dependent manner with 25-50 mg kg(-1) of gefitinib administrated orally. Inhibition of EGFR-transmitted growth stimulation by gefitinib was clearly observed in anaplastic thyroid cancer cell lines. Our results suggested that EGFR targeted therapy, such as gefitinib, might be worth further investigation for the treatment of anaplastic thyroid cancer. PMID- 15785739 TI - Frequencies of HER-2/neu expression and gene amplification in patients with oesophageal squamous cell carcinoma. AB - The utilisation of antitumour T cells induced by cancer vaccination with HER-2 peptides or antibodies (Herceptin) against HER-2, as immunotherapy for oesophageal cancer, is a novel and attractive approach. It is important to clarify the frequencies of HER-2 expression and gene amplification in patients with oesophageal squamous cell carcinoma (SCC) and to evaluate the relationship between HER-2 status and HLA haplotype, since the candidates for HER-2 peptide based vaccination are restricted to a certain HLA haplotype. We determined the frequency of HER-2 expression using the HercepTest for immunohistochemistry and HER-2 gene amplification by fluorescence in situ hybridisation (FISH) assay in oesophageal SCC (n=66). HER-2-positive tumours (1+/2+/3+) analysed by a HercepTest were observed in 30.3% of all the patients and HER-2 gene amplification evaluated by FISH was observed in 11.0% of all the patients, in which all HercepTest (3+) tumours were found to have gene amplification and three of six moderately positive (2+) tumours showed gene amplification. Furthermore, HER-2-positive cells were present more diffusely and were larger within each tumour in the patients who were HercepTest 3+ than those who were HercepTest 1+. Moreover, the survival rate in HER-2-positive group was significantly worse than that in HER-2-negative group. Also, the survival rate in the patients with HER-2 gene amplification was significantly worse than that without HER-2 gene amplification. In addition, oesophageal SCC patients with both HLA-A24-positive and HER-2-positive tumours (1+/2+/3+) accounted for 26% of these cases, and both HLA-A2- and HER-2-positive tumours accounted for 18% of them. PMID- 15785740 TI - Mitochondrial D-loop mutations and deletion profiles of cancerous and noncancerous liver tissue in hepatitis B virus-infected liver. AB - The largest single underlying cause of hepatocellular carcinoma (HCC) worldwide is hepatitis B virus (HBV) infection. Hepatitis B virus increases cellular oxidative stress and the development of HCC occurs after a long latency period. The study was carried out to determine whether mitochondrial DNA abnormalities were associated with HCC in individuals with HBV. The frequency of mutation and deletion of specific areas of the mitochondrial genome in tumour and matched normal tissue of patients with HBV infection was investigated in the current study. The percentage of control subjects harbouring D-loop mutations was 11%, which was significantly lower than that observed in both the noncancerous (49%, P=0.033) and tumour tissue (59%, P=0.014) of patients with HCC. In contrast, the number of cases in which the common 4977 bp deletion of the mitochondrial genome was detected was significantly greater in control liver and noncancerous liver tissue of subjects with HCC (100 and 95%, respectively) than in cancerous liver tissue (28%, P<0.001). These observations suggest that the inflammatory process contributes to the rate of mitochondrial mutations. However, the lower frequency of the large deletion in cancerous tissue suggests that there is selection against either mitochondria, which harbour large deletions, or against cells that contain these mitochondria during hepatocarcinogenesis. PMID- 15785741 TI - Galectin-1 as a potential cancer target. AB - Galectins are a family of structurally related carbohydrate-binding proteins, which are defined by their affinity for poly-N-acetyllactosamine-enriched glycoconjugates and sequence similarities in the carbohydrate recognition domain. Galectin-1, a member of this family, contributes to different events associated with cancer biology, including tumour transformation, cell cycle regulation, apoptosis, cell adhesion, migration and inflammation. In addition, recent evidence indicates that galectin-1 contributes to tumour evasion of immune responses. Given the increased interest of tumour biologists and clinical oncologists in this field and the potential use of galectins as novel targets for anticancer drugs, we summarise here recent advances about the role of galectin-1 in different events of tumour growth and metastasis. PMID- 15785742 TI - Intensified concurrent chemoradiotherapy with 5-fluorouracil and irinotecan as neoadjuvant treatment in patients with locally advanced rectal cancer. AB - This study aimed to evaluate the feasibility and efficacy of neoadjuvant chemoradiotherapy intensified with irinotecan in patients with locally advanced rectal cancer. Eligible patients had nonmetastatic disease at a locally advanced stage that made R0 resection and sphincter preservation uncertain. They received preoperative radiation over 6 weeks to 45 Gy and boost of 5.4 Gy and concurrent continuous infusion 5-fluorouracil 250 mg m(-2) day(-1) and weekly irinotecan 40 mg m(-2). In all, 37 patients entered the study. T stage at baseline as determined by ultrasound was T2/T3/T4 in 2/19/16 patients; 31 patients had lymph node involvement. The predominant toxicity was diarrhoea (grade 3/4 in 10/2 patients). Haematologic toxicity and surgical complications were moderate. Among 36 patients undergoing surgery, 32 (89%) had R0 resection and 23 (64%) sphincter preservation. Pathologic complete response (pCR) was achieved in eight (22%) of 36 patients, and 10 patients (28%) had only microscopic residual disease. At 4 years, overall survival was 66%, disease-free survival 73%, local relapse rate 7%, and distant failure rate 24%. Extent of resection and postoperative nodal status were significant predictors of overall and disease-free survival. Intensified neoadjuvant chemoradiotherapy with irinotecan can be safely administered and results in a high pCR rate. PMID- 15785743 TI - Remarks on the BOADICEA model of genetic susceptibility to breast and ovarian cancer. PMID- 15785745 TI - Preoperative short-term radiation therapy (25 Gy, 2.5 Gy twice daily) for primary resectable rectal cancer (phase II). AB - To evaluate the feasibility, effectiveness, and long-term bowel function of preoperative hyperfractionated accelerated radiotherapy in primary resectable rectal cancer. A total of 184 consecutive patients (median age 65 years, male : female=2 : 1) with clinical T3Nx rectal adenocarcinoma received preoperative pelvic radiation therapy with single fractions of 2.5 Gy twice daily (interval 6 h between fractions) to a total dose of 25 Gy within 1 week. Surgery was conducted the following week. Postoperative histology revealed UICC stage I in 33%, stage II in 26%, stage III in 34%, and stage IV in 7% of the patients. Median follow-up was 43 months (53 months for surviving patients). The actuarial 4-year-local-recurrence rate was 2.1%, overall recurrence 23%. Disease-specific and disease-free survivals at 4 years (excluding stage IV) were 82 and 69%, respectively. Overall survival for 4 years was 68%. Postoperative mortality was 0.5% (one patient), early anastomotic leakage occurred in 11.4%, and anastomotic stenosis requiring treatment in 6%, of 132 patients with primary anastomosis. Seven of 184 patients (3.8%) died of abdominal complications, all within the first year. Bowel function was satisfactory after more than 5 years. Local control in primarily resectable rectal cancer after 10 x 2.5 Gy is excellent, warranting further evaluation of this treatment. PMID- 15785746 TI - Blockage of angiotensin II type I receptor decreases the synthesis of growth factors and induces apoptosis in C6 cultured cells and C6 rat glioma. AB - Angiotensin II (Ang II) is a main effector peptide in the renin-angiotensin system and participates in the regulation of vascular tone. It also has a role in the expression of growth factors that induce neovascularisation which is closely associated to the growth of malignant gliomas. We have shown that the selective blockage of the AT1 receptor of angiotensin inhibits tumour growth, cell proliferation and angiogenesis of C6 rat glioma. The aim of this study was to study the effects of the blockage of AT1 receptor on the synthesis of growth factors, and in the genesis of apoptosis in cultured C6 glioma cells and in rats with C6 glioma. Administration of losartan at doses of 40 or 80 mg kg(-1) to rats with C6 glioma significantly decreased tumoral volume and production of platelet derived growth factor, vascular endothelial growth factor and basic fibroblast growth factor. It also induced apoptosis in a dose-dependent manner. Administration of Ang II increased cell proliferation of cultured C6 cells which decreased by the administration of losartan. Our results suggest that the selective blockage of AT1 diminishes tumoral growth through inhibition of growth factors and promotion of apoptosis. PMID- 15785747 TI - Clinical application of biological markers for treatments of resectable non-small cell lung cancers. AB - We performed a clinical study to identify biological markers useful for the treatment of resectable non-small-cell lung cancers (NSCLCs). In all, 173 patients were studied. By immunohistochemistry, we evaluated the Ki-67 proliferation index, tumour vascularity, thymidylate synthase (TS), vascular endothelial growth factor (VEGF)-A, VEGF-C, and E (epithelial)-cadherin. Concerning the survival of NSCLC patients, tumour vascularity (P<0.01), VEGF-A status (P=0.03), VEGF-C status (P=0.03), and E-cadherin status (P=0.03) were significant prognostic factors in patients with stage I NSCLCs. The Ki-67 proliferation index (P=0.02) and TS status (P<0.01) were significant prognostic factors in patients with stage II-III NSCLCs. In patients with stage II-III NSCLCs, furthermore, the survival of UFT (a combination of tegafur and uracil) treated patients with TS-negative tumours was significantly better than those of any other patients. Biological markers associated with tumour angiogenesis or metastasis are useful for the detection of aggressive tumours among early-stage NSCLCs. Postoperative chemotherapy might be necessary in such tumours even in stage I. In contrast, tumour proliferation rate and TS status are useful markers for identifying less aggressive tumours in locally advanced NSCLCs. Thymidylate synthase expression is also a useful marker to evaluate responsiveness of UFT based chemotherapy for these tumours. PMID- 15785748 TI - Gene expression fingerprint of uterine serous papillary carcinoma: identification of novel molecular markers for uterine serous cancer diagnosis and therapy. AB - Uterine serous papillary cancer (USPC) represents a rare but highly aggressive variant of endometrial cancer, the most common gynecologic tumour in women. We used oligonucleotide microarrays that interrogate the expression of some 10 000 known genes to profile 10 highly purified primary USPC cultures and five normal endometrial cells (NEC). We report that unsupervised analysis of mRNA fingerprints readily distinguished USPC from normal endometrial epithelial cells and identified 139 and 390 genes that exhibited >5-fold upregulation and downregulation, respectively, in primary USPC when compared to NEC. Many of the genes upregulated in USPC were found to represent adhesion molecules, secreted proteins and oncogenes, such as L1 cell adhesion molecule, claudin-3 and claudin 4, kallikrein 6 (protease M) and kallikrein 10 (NES1), interleukin-6 and c-erbB2. Downregulated genes in USPC included SEMACAP3, ras homolog gene family, member I (ARHI), and differentially downregulated in ovarian carcinoma gene 1. Quantitative RT-PCR was used to validate differences in gene expression between USPC and NEC for several of these genes. Owing to its potential as a novel therapeutic marker, expression of the high-affinity epithelial receptor for Clostridium perfringens enterotoxin (CPE) claudin-4 was further validated through immunohistochemical analysis of formalin-fixed paraffin-embedded specimens from which the primary USPC cultures were obtained, as well as an independent set of archival USPC specimens. Finally, the sensitivity of primary USPC to the administration of scalar doses of CPE in vitro was also demonstrated. Our results highlight the novel molecular features of USPC and provide a foundation for the development of new type-specific therapies against this highly aggressive variant of endometrial cancer. PMID- 15785749 TI - NK105, a paclitaxel-incorporating micellar nanoparticle formulation, can extend in vivo antitumour activity and reduce the neurotoxicity of paclitaxel. AB - Paclitaxel (PTX) is one of the most effective anticancer agents. In clinical practice, however, high incidences of adverse reactions of the drug, for example, neurotoxicity, myelosuppression, and allergic reactions, have been reported. NK105, a micellar nanoparticle formulation, was developed to overcome these problems and to enhance the antitumour activity of PTX. Via the self-association process, PTX was incorporated into the inner core of the micelle system by physical entrapment through hydrophobic interactions between the drug and the well-designed block copolymers for PTX. NK105 was compared with free PTX with respect to their in vitro cytotoxicity, in vivo antitumour activity, pharmacokinetics, pharmacodynamics, and neurotoxicity. Consequently, the plasma area under the curve (AUC) values were approximately 90-fold higher for NK105 than for free PTX because the leakage of PTX from normal blood vessels was minimal and its capture by the reticuloendothelial system minimised. Thus, the tumour AUC value was 25-fold higher for NK105 than for free PTX. NK105 showed significantly potent antitumour activity on a human colorectal cancer cell line HT-29 xenograft as compared with PTX (P<0.001) because the enhanced accumulation of the drug in the tumour has occurred, probably followed by its effective and sustained release from micellar nanoparticles. Neurotoxicity was significantly weaker with NK105 than with free PTX. The neurotoxicity of PTX was attenuated by NK105, which was demonstrated by both histopathological (P<0.001) and physiological (P<0.05) methods for the first time. The present study suggests that NK105 warrants a clinical trial for patients with metastatic solid tumours. PMID- 15785750 TI - The role of dendritic cell precursors in tumour vasculogenesis. AB - In this review, we discuss the recent identification in vivo of a population of CD11c+ cells exhibiting simultaneous expression of both endothelial and dendritic cell markers, termed vascular leukocytes (VLCs). VLCs are highly represented in human ovarian carcinomas and, depending on the milieu, can assemble into functional blood vessels or act as antigen-presenting cells. The identification of dendritic cell precursors as bipotent cells has important implications for the physiopathology and therapy of tumours. VLCs emerge as a novel therapeutic target against tumour vascularisation. PMID- 15785751 TI - Hormone use for menopausal symptoms and risk of breast cancer. A Danish cohort study. AB - Numerous studies and meta-analyses have shown that hormone replacement therapy (HRT) for menopausal symptoms increases the risk of developing breast cancer, estimated to be 2.3% for each year of use. The influence of different oestrogen progestin regimens has still not been fully evaluated. Using longitudinal data from the population-based prescription database of the county of North Jutland, Denmark, and the Danish Cancer Registry, we examined the risk of developing breast cancer in relation to HRT in a cohort of 78,380 women aged 40-67 years from 1989 to 2002. A total of 1462 cases of breast cancer were identified during a mean follow-up of 10 years. Use of HRT did not increase the risk of breast cancer in women aged 40-49 years. Restricting the cohort to 48,812 women aged 50 years or more at entry, of whom 15 631 were HRT users, we found an increased risk associated with current use of HRT (relative risk 1.61, 95% confidence interval 1.38-1.88). The risk increased with increasing duration of use and decreased with time since last HRT prescription, reaching unity after 5 years. No material risk difference was observed among the various HRT-regimens. This population-based cohort study provides further confirmation that HRT increases the risk of developing breast cancer in women aged 50 years or more. PMID- 15785752 TI - Waiting times for radiotherapy: variation over time and between cancer networks in southeast England. AB - The aim of this study was to investigate variations in the length of time that patients with cancer wait from diagnosis to treatment with radiotherapy. A total of 57,426 men and 71,018 women diagnosed with cancer between 1992 and 2001 and receiving radiotherapy within 6 months of diagnosis were identified from the Thames Cancer Registry database. In total, 12 sites were identified for which a substantial number or proportion of patients received radiotherapy: head and neck, oesophagus, colon, rectum, lung, nonmelanoma skin cancer, breast, uterus, prostate, bladder, brain and non-Hodgkin's lymphoma. Median waiting times from diagnosis to radiotherapy were calculated, together with the proportion of patients who received radiotherapy within 60 days of diagnosis, and analysed by year of diagnosis, cancer site, deprivation quintile, age at diagnosis, sex and cancer network of either residence or treatment. Logistic regression was used to adjust the proportion receiving treatment within 60 days for the effects of the other factors. There were significant differences in the proportions receiving radiotherapy within 60 days between different networks and different cancer sites, which remained after adjustment. Median waiting times varied from 42 to 65 days across networks of residence, with the adjusted proportion treated within 60 days ranging from 44 to 71%. There was no difference between male and female patients after adjustment for the other factors, particularly site. There was a highly significant trend over time: the median wait increased from 45 days in 1992 to 76 days in 2001, while the adjusted proportion being treated within 60 days declined by almost a half, from 64 to 35%, over the same period. PMID- 15785754 TI - Recent trends in cancer mortality in the UK. AB - Comparisons of survival rates of given diseases with different treatments or in different places often gave misleading results until the introduction of controlled trials. Recent reports of relatively low survival rates following the treatment of cancer in the UK compared to the rates in other countries, not based on controlled trials, may consequently be misleading. Their validity has, therefore, been tested by comparing the levels and trends in mortality--the ultimate criterion by which the success or failure of any system of care can be judged. For this purpose, rates and trends in rates over 20-50 years have been compared in five European countries of similar economic status (France, Italy, the Netherlands, Sweden, and the UK). The UK rates are not generally worse than those in the other countries and are sometimes better. Exceptions were cancer of the lung, large bowel, and breast, the first of which is explained by differences in the prevalence of smoking. PMID- 15785753 TI - Genomic and gene expression profiling of minute alterations of chromosome arm 1p in small-cell lung carcinoma cells. AB - Genetic alterations occurring on human chromosome arm 1p are common in many types of cancer including lung, breast, neuroblastoma, pheochromocytoma, and colorectal. The identification of tumour suppressors and oncogenes on this arm has been limited by the low resolution of current technologies for fine mapping. In order to identify genetic alterations on 1p in small-cell lung carcinoma, we developed a new resource for fine mapping segmental DNA copy number alterations. We have constructed an array of 642 ordered and fingerprint-verified bacterial artificial chromosome clones spanning the 120 megabase (Mb) 1p arm from 1p11.2 to p36.33. The 1p arm of 15 small-cell lung cancer cell lines was analysed at sub-Mb resolution using this arm-specific array. Among the genetic alterations identified, two regions of recurrent amplification emerged. They were detected in at least 45% of the samples: a 580 kb region at 1p34.2-p34.3 and a 270 kb region at 1p11.2. We further defined the potential importance of these genomic amplifications by analysing the RNA expression of the genes in these regions with Affymetrix oligonucleotide arrays and semiquantitative reverse transcriptase polymerase chain reaction. Our data revealed overexpression of the genes HEYL, HPCAL4, BMP8, IPT, and RLF, coinciding with genomic amplification. PMID- 15785755 TI - Gene expression profiling of colorectal adenomas and early invasive carcinomas by cDNA array analysis. AB - It is generally accepted that most colorectal carcinomas arise in pre-existing adenomas. Morphologically, colorectal adenomas can be divided into two groups, protruded type and flat type. The aim of this study was to clarify relevant alterations of gene expression associated with the early stage of colorectal carcinogenesis. Using cDNA array, we analysed the expression profiles of 550 cancer-related genes in 36 colorectal adenomas (18 flat-type and 18 protruded type adenomas) and 14 early invasive carcinomas. Among the 550 genes, we chose 32 genes the average expression levels of which were at least three-fold up- or downregulated in tumour tissues compared with levels in matched normal tissues. A total of 13 and 19 genes were identified as up- and downregulated genes in tumour tissues, respectively. Among the upregulated genes, the average expression levels of E1AF, bone morphogenic protein (BMP)-4, insulin-like growth factor (IGF)-2, inducible nitric oxide synthase (iNOS), tissue inhibitors of metalloproteinase (TIMP)-1, Smad4, and nm23 in tumour tissues were over five times higher than those in matched normal tissues. Colorectal adenomas and early invasive carcinomas were divided into two major clusters by clustering analysis. Moreover, flat- and protruded-type adenomas were divided into two major clusters by clustering analysis. The expression profiles obtained by the cDNA array clearly indicate that colorectal adenomas and early invasive carcinomas have specific expression profiles. Likewise, the gene expression profiles of flat- and protruded-type adenomas are different. These results indicate that molecular classification of early colorectal tumours by a cDNA array is feasible. PMID- 15785756 TI - Essence of harmony. PMID- 15785757 TI - Natural regulatory T cells and self-tolerance. AB - The adaptive immune system allows individual organisms to mount defensive reactions against unanticipated pathogens by developmentally creating a diverse repertoire of clonally distributed receptors capable of recognizing a multitude of antigens and then expanding as effector cell populations those that can recognize molecules from the pathogens. To function properly, the system must deal with the problem of randomly generated receptors that can recognize self components. Most solutions to this self-tolerance problem are cell intrinsic and involve the deletion or inactivation of autoreactive cells. However, an extrinsic form of dominant tolerance has been demonstrated that takes the form of CD4(+) regulatory T cells. This perspective discusses why such a mechanism might have evolved and the problems it presents for self-non-self discrimination. PMID- 15785758 TI - A well adapted regulatory contrivance: regulatory T cell development and the forkhead family transcription factor Foxp3. AB - The random generation of antigen receptors in developing lymphocytes results in a considerable risk of autoimmunity. Regulatory T cells (T(reg) cells) act in a dominant, trans-acting way to actively suppress immune activation and maintain immune tolerance. Here, we discuss the principal advances in our understanding of the molecular mechanisms of T(reg) cell development and function with particular emphasis on the forkhead transcription factor Foxp3. Accumulating evidence suggests that T(reg) cells represent a dedicated T cell lineage and that Foxp3 functions as the T(reg) cell lineage specification factor. The aggressive early onset lymphoproliferative syndrome resulting from Foxp3 deficiency identifies T(reg) cells as vital mediators of immunological tolerance to self and Foxp3 as the mediator of the genetic mechanism of dominant tolerance. PMID- 15785759 TI - Mechanisms of suppression by suppressor T cells. AB - Mechanisms of immunosuppression by CD4(+)CD25(+) suppressor T cells have been addressed using many in vitro and in vivo conditions. However, those studies have not yielded a single mode of action. This review will discuss the mechanisms of suppression, which include the local secretion of cytokines such as TGF-beta and direct cell contact through binding of cell surface molecules such as CTLA-4 on suppressor T cells to CD80 and CD86 molecules on effector T cells. Suppression requires the appropriate colocalization of suppressor and effector T cells in different tissue and may involve the interference with T cell receptor signaling that triggers transcription factors important in regulating effector cell function. PMID- 15785760 TI - Naturally arising Foxp3-expressing CD25+CD4+ regulatory T cells in immunological tolerance to self and non-self. AB - Naturally arising CD25(+)CD4(+) regulatory T cells actively maintain immunological self-tolerance. Deficiency in or dysfunction of these cells can be a cause of autoimmune disease. A reduction in their number or function can also elicit tumor immunity, whereas their antigen-specific population expansion can establish transplantation tolerance. They are therefore a good target for designing ways to induce or abrogate immunological tolerance to self and non-self antigens. PMID- 15785761 TI - Natural regulatory T cells in infectious disease. AB - This review discusses the control exerted by natural CD4(+) CD25(+) regulatory T cells (natural T(reg) cells) during infectious processes. Natural T(reg) cells may limit the magnitude of effector responses, which may result in failure to adequately control infection. However, natural T(reg) cells also help limit collateral tissue damage caused by vigorous antimicrobial immune responses. We describe here various situations in which the balance between natural T(reg) cells and effector immune functions influences the outcome of infection and discuss how manipulating this equilibrium might be exploited therapeutically. PMID- 15785763 TI - Switching sides: transcriptional control of CD4 thymocyte lineage commitment. PMID- 15785764 TI - Favorite flavors of surfaces. PMID- 15785765 TI - Inflammatory chemokines: tuned in, turned on, dropped out. PMID- 15785766 TI - MAPK p38: alternative and nonstressful in T cells. PMID- 15785769 TI - Infrared radiation from an extrasolar planet. AB - A class of extrasolar giant planets--the so-called 'hot Jupiters' (ref. 1)--orbit within 0.05 au of their primary stars (1 au is the Sun-Earth distance). These planets should be hot and so emit detectable infrared radiation. The planet HD 209458b (refs 3, 4) is an ideal candidate for the detection and characterization of this infrared light because it is eclipsed by the star. This planet has an anomalously large radius (1.35 times that of Jupiter), which may be the result of ongoing tidal dissipation, but this explanation requires a non-zero orbital eccentricity (approximately 0.03; refs 6, 7), maintained by interaction with a hypothetical second planet. Here we report detection of infrared (24 microm) radiation from HD 209458b, by observing the decrement in flux during secondary eclipse, when the planet passes behind the star. The planet's 24-microm flux is 55 +/- 10 microJy (1sigma), with a brightness temperature of 1,130 +/- 150 K, confirming the predicted heating by stellar irradiation. The secondary eclipse occurs at the midpoint between transits of the planet in front of the star (to within +/- 7 min, 1sigma), which means that a dynamically significant orbital eccentricity is unlikely. PMID- 15785770 TI - Genome-wide non-mendelian inheritance of extra-genomic information in Arabidopsis. AB - A fundamental tenet of classical mendelian genetics is that allelic information is stably inherited from one generation to the next, resulting in predictable segregation patterns of differing alleles. Although several exceptions to this principle are known, all represent specialized cases that are mechanistically restricted to either a limited set of specific genes (for example mating type conversion in yeast) or specific types of alleles (for example alleles containing transposons or repeated sequences). Here we show that Arabidopsis plants homozygous for recessive mutant alleles of the organ fusion gene HOTHEAD (HTH) can inherit allele-specific DNA sequence information that was not present in the chromosomal genome of their parents but was present in previous generations. This previously undescribed process is shown to occur at all DNA sequence polymorphisms examined and therefore seems to be a general mechanism for extra genomic inheritance of DNA sequence information. We postulate that these genetic restoration events are the result of a template-directed process that makes use of an ancestral RNA-sequence cache. PMID- 15785771 TI - Prospective evaluation of gut mucosal barrier injury following various myeloablative regimens for haematopoietic stem cell transplant. AB - We determined gut mucosal barrier injury (MBI) among 129 recipients of an allogeneic or autologous haematopoietic stem cell transplant (HSCT) who had been given different myeloablative regimens by measuring integrity using the lactulose/rhamnose (RHA) ratio and absorption using the ratios of rhamnose/3-O methylglucose and xylose/3-O-methylglucose. Regimens that did not contain idarubicin induced oral mucositis and disturbed gut integrity and absorption earlier than did those containing the anthracycline. By contrast, regimens containing idarubicin induced more severe and prolonged oral and gut MBI. Gut integrity and absorption of most patients were still abnormal at discharge from hospital. These results confirm that the integrity and absorptive capacity of the gut is affected adversely by myeloablative regimens in general, although only two patterns of mucosal injury emerged depending on whether or not idarubicin was used. PMID- 15785772 TI - Inefficiency of high-dose G-CSF alone as second mobilization regimen in fludarabin-cyclophosphamide-treated CLL patients who failed to mobilize after chemotherapy and G-CSF. PMID- 15785773 TI - Dietary patterns and lifestyle factors in the Norwegian EPIC cohort: the Norwegian Women and Cancer (NOWAC) study. AB - OBJECTIVE: To identify different dietary patterns in Norway using a combination of cluster and factor analysis. DESIGN: Cross-sectional study. SETTING: Nation wide, population-based study. SUBJECTS: The Norwegian EPIC cohort is a subcohort of the Norwegian Women and Cancer study (NOWAC), and consist 37.226 women aged 41 56 y who answered a food frequency questionnaire (FFQ) in 1998. INTERVENTIONS: The associations among 50 food variables were first investigated by using principal component analysis. Five important factors were found. The five principal components were then used as input in the cluster analysis. Different socioeconomic and lifestyle variables were examined. RESULTS: Six clusters of dietary patterns were found, and were labelled accordingly: 'traditional fish eaters', 'healthy eaters', 'average, less fish, less healthy', 'Western', 'traditional bread eaters', and 'alcohol users'. The traditional fish eaters and the traditional bread eaters were both highly represented in the north and west of Norway and were more likely to be present among persons with lower income and lower education. The healthy and the alcohol drinkers were found mostly in the south and east and were more likely to have higher income. Persons in the alcohol group were more likely to be current smokers. The western group had the highest percentage of three or more persons in the household and the shortest time since last birth, indicating that families with children dominate this group. CONCLUSION: Our data indicate six different dietary patterns in Norway, each with different socio-demographic and lifestyle characteristics. SPONSORSHIP: The Norwegian Cancer Society (E 04038/006). PMID- 15785774 TI - Interaction of the LMX1B and PAX2 gene products suggests possible molecular basis of differential phenotypes in Nail-Patella syndrome. AB - The LMX1B gene, encoding a protein involved in limb, kidney and eye development, is mutated in patients affected by Nail-Patella syndrome. Inter- and intrafamilial variability is common in this disorder for skeletal abnormalities, presence and severity of nephropathy and ocular anomalies. Phenotypic variability might depend on interactions of the LMX1B causative gene with other genes during development of both kidney and eye, which might act as modifier genes. Results are presented on the interaction between LMX1B and PAX2 proteins, obtained by both direct yeast two-hybrid assay and coimmunoprecipitation. Such interaction provides support to further studies on pathways underlying important developmental processes. PMID- 15785775 TI - Complex trait mapping in isolated populations: Are specific statistical methods required? AB - In this paper, we review the statistical methods that can be used in isolated populations to map genes involved in complex diseases. Our intention is to highlight the fact that if the features of population isolates may help in the identification of susceptibility factors for complex traits, the choice and design of methods for statistical analysis in these populations deserve particular care. We show that methods designed for outbred samples are generally not appropriate for isolated populations and could lead to false conclusions. PMID- 15785776 TI - Genetic stratification of pathogen-response-related and other variants within a homogeneous Caucasian Irish population. AB - Selection pressures from pathogens impact on the worldwide geographic distribution of polymorphisms in certain pathogen-response-associated genes. Such gene-specific effects could lead to confounding by geographic disease associations. We wished to determine if such constraints impinge on the genetic structure of a population of Irish patients and whether variants associated with responses to pathogens showed greater stratification. The counties of origin of each subject's grandparents were used as the geographic variable. F(st), proportional to the extent of population structure, was low (mean F(st)=0.004 across 25 SNPs, range 0.001-0.008) and it was not significantly higher for pathogen response SNPs (F(st)=0.004) than for other SNPs (F(st)=0.003, P=0.21). Correspondence analysis revealed weak trends primarily in approximately northeast to southwest and secondarily in northwest to southeast directions. One dimensional spatial autocorrelation analysis revealed a weak (Moran's I autocorrelation of -0.10) tendency for SNP frequencies to diverge with greater distance. Two-dimensional autocorrelation indicated a northeast to southwest gradient that was similar for both the pathogen response and other SNPs. The southeastern county, Wexford, showed a distinctive pattern, perhaps consistent with Anglo-Norman settlements. In conclusion, these results indicate that pathogen response SNPs do not exhibit significantly more population structure than other SNPs within this Caucasian population. This suggests that the specific population structure of particular genes may not typically be a cause of strong confounding in genetic studies where population structure is controlled. PMID- 15785777 TI - Kabuki syndrome: new ocular findings but no evidence of 8p22-p23.1 duplications in a clinically defined cohort. AB - The underlying cause of the multiple congenital anomalies/mental retardation syndrome Kabuki syndrome (KS, OMIM 147920) has not yet been established. We identified seven patients who fulfilled the classical clinical criteria for this syndrome and undertook a detailed clinical, ophthalomological and molecular cytogenetic review. Three of the seven patients had previously undetected ocular anomalies including myopia, ptosis, strabismus and tilted discs. The identification of preventable causes of loss of vision underlines the value of detailed ophthalmologic examination of KS patients. Using BAC fluorescence in situ hybridisation, there was no evidence of the duplication of 8p recently reported by Milunsky and Huang. We conclude that the cause of KS has yet to be established. PMID- 15785778 TI - Environmental exposure to trace elements and risk of cutaneous melanoma. AB - PURPOSE: Our aim was to examine the risk of melanoma in association with exposure to trace elements of toxicological and nutritional interest. METHODS: We analyzed the concentrations of cadmium, lead, chromium, selenium, copper and zinc in toenails of 58 patients with newly diagnosed cutaneous melanoma as well as in 58 age- and sex-matched control subjects, randomly selected from the population of Modena province in northern Italy. RESULTS: Melanoma risk was substantially unrelated to toenail levels of cadmium, chromium, lead and selenium. Subjects with higher toenail copper levels showed an excess risk, both in the crude analysis and after adjusting for sun exposure and level of education, while in both analyses high iron concentrations were associated with a decreased risk of the disease. A weak direct association between zinc levels and melanoma risk also emerged in the multivariate analysis. CONCLUSIONS: Overall, these results do not suggest an involvement of heavy metals in melanoma etiology, while they do give some support to a possible role of zinc and, in particular, copper and iron exposure in influencing disease risk. However, these findings must be evaluated with caution due to the limited statistical stability of the point estimates. PMID- 15785780 TI - Risk of becoming cocaine dependent: epidemiological estimates for the United States, 2000-2001. AB - In this paper, we present new estimates for the risk of becoming cocaine dependent within 24 months after first use of the drug, and study subgroup variation in this risk. The study estimates are based on the National Household Survey on Drug Abuse conducted during 2000-2001, with a representative sample of US residents aged 12 years and older (n=114 241). A total of 1081 respondents were found to have used cocaine for the first time within 24 months prior to assessment. Between 5 and 6% of these recent-onset users had become cocaine dependent since onset of use. Less [corrected] risk of recent cocaine dependence soon after onset of cocaine use was found for female subjects, young adults aged 21-25 years, and non-Hispanic Black/African-Americans. Use of crack-cocaine and taking cocaine by injection were associated with having become cocaine dependent soon after onset of use. These epidemiologic findings help to quantify the continuing public health burden associated with new onsets of cocaine use in the 21st century. PMID- 15785779 TI - Ethanol, 3,4-methylenedioxymethamphetamine (ecstasy) and their combination: long term behavioral, neurochemical and neuropharmacological effects in the rat. AB - This study investigated long-term behavioral, neurochemical, and neuropharmacological effects of ethanol-(+/-)-3,4-methylenedioxymethamphetamine (MDMA, ecstasy) combinations. Over 4 consecutive days, male Long-Evans rats received 1.5 g/kg ethanol and/or 10 mg/kg MDMA, or saline. Rectal temperatures were taken in some rats. Starting 4 days after the last injection, we tested working memory, sensory-motor coordination, and anxiety. Subsequently, we measured cortical, striatal, septal, and hippocampal monoamines (last MDMA injection-euthanasia delay: 20 days), or electrically evoked release of serotonin (5-HT) in cortical and hippocampal slices, and its modulation in the presence of CP 93,129 (3-(1,2,5,6-tetrahydropyrid-4-yl)pyrrollo[3,2-b]pyrid-5-one) or methiotepin (last MDMA injection-euthanasia delays: 3-6 weeks). Ethanol attenuated the MDMA-induced hyperthermia, but only on the first day. In the long term, MDMA reduced 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) content in most brain regions. The behavioral and neurochemical effects of the ethanol-MDMA combination were comparable to those of MDMA alone; sensory-motor coordination was altered after ethanol and/or MDMA. In hippocampal slices from rats given ethanol and MDMA, the CP 93,129-induced inhibition and methiotepin-induced facilitation of 5-HT release were stronger and weaker, respectively, than in the other groups. This is the first study addressing long-term effects of repeated MDMA and EtOH combined treatments in experimental animals. Whereas the drug combination produced the same behavioral and neurochemical effects as MDMA alone, our neuropharmacological results suggest that MDMA-EtOH interactions may have specific long-term consequences on presynaptic modulation of hippocampal 5-HT release, but not necessarily related to MDMA-induced depletion of 5-HT. Thus, it is likely that the psycho(patho)logical problems reported by ecstasy users drinking alcohol are not solely due to the consumption of MDMA. PMID- 15785781 TI - Host-associated populations in the lettuce root aphid, Pemphigus bursarius (L.). AB - Pemphigus bursarius is a host-alternating aphid in which annual rounds of sexual reproduction on its primary host, Populus nigra, are interspersed with parthenogenesis on a range of secondary hosts. Evidence was sought for the existence of genetically distinct populations, associated with different secondary hosts, in P. bursarius. Microsatellite markers revealed that genetically distinct populations were present on three different secondary host species. Microsatellites were also used, in conjunction with mitochondrial DNA sequence variation, to investigate the relationships between aphids on Populus, following sexual reproduction, and those on the secondary hosts. Evidence was found for a distinct, cyclically parthenogenetic population that exploited Lactuca sativa as its secondary host. In contrast, populations associated with Matricaria inodora appeared to be largely composed of obligate parthenogens or may even have been another species of Pemphigus. Populations on Lapsana communis appeared to be a mixture of cyclical and obligate parthenogens and were more genetically heterogeneous than those on other secondary hosts, possibly due to founder effects. Experiments to measure the performance of P. bursarius clones on different secondary hosts were inconclusive, failing to demonstrate either the presence or absence of adaptations to secondary hosts. PMID- 15785782 TI - A secondary hybrid zone between diploid Dactylorhiza incarnata ssp. cruenta and allotetraploid D. lapponica (Orchidaceae). AB - Secondary hybrid zones are not uncommon in Dactylorhiza, but knowledge of ecological and evolutionary consequences of hybridization are scarce. Here, we assess interploidal gene flow and introgression in a hybrid zone between diploid Dactylorhiza incarnata ssp. cruenta (2n = 2x = 40) and its putative allotetraploid derivative D. lapponica (2n = 4x = 80). Photometric quantification of DNA content and morphology confirmed that triploids are abundant in sympatric populations in our study area. Allozyme segregation patterns in D. lapponica supported an allopolyploid origin, although unbalanced genotypes suggested rare pairings between homoeologous chromosomes. Photometric data and chromosome counts suggest backcrossing between the triploid hybrid and D. lapponica, and hence some hybrid fertility. Triploids are morphologically more similar to the tetraploids than the diploids, maybe owing to the hybrid origin of both triploids and tetraploids. The diploids and tetraploids were not more similar in the parapatric populations compared to when they occur in allopatry. This indicates that backcrossing rarely leads to introgression, or alternatively that allopatric populations are not isolated enough to prevent influx of pollen from the other species. Despite some evidence of backcrossing, our study gives few indications that widespread hybridization entails local breakdown of species boundaries. Rather, the hybrid zone may be a transient phenomenon due to intensive mowing, resulting in the opening of habitats and hence bringing the parental species into close contact. PMID- 15785783 TI - [Problems and resources among hospital physicians, residents, and nurses in their hospital work]. AB - Research about fieldwork with 133 health professionals from different hospitals in Buenos Aires city about both functional and disfunctional discomfort perceived and expressed by them. Risk factors/items described are: work in isolation, naturalizing of discomfort, overload, overadaptation, inappropriate relieving methods: tobacco, alcohol, too much or insufficient food, anxiety, self - carelessness, both physical or emotional, unawareness of their rights as workers. Work in hospitals also provides satisfaction. Some questions and reflexions are stated and a minimum decalogue of measures to prevent and promote health in hospital workers is proposed. PMID- 15785784 TI - [New assessment variables in the Clock Drawing Test (CDT) allow an early and easy diagnosis of degrees of cognitive impairment]. AB - OBJECTIVE: We developed new subroutines and scoring procedures of the CDT to detect markers of early cognitive impairment. SUBJECTS AND METHODS: We compared the performance of 41 mild Alzheimer's patients (AD) (MMSE 22.7+/-3.2); and 18 mild cognitive impairment patients (MCI) (MMSE 28.3+/-1.4), with 33 age and education matched normal controls (NC) regarding their ability in drawing a clock on command, copying a printed clock, setting hands and reading the time. We search for differences in the placement of the hands using a colloquial hour code ("a quarter to ten") and a formal numeric code ("10:45"). We focused our attention on placement and clock hands size. The drawing of hands pointing at 2:50 hs and 8:40 hs were of particular interest. RESULTS: Planning strategies, placement of clock hands using a formal numeric code, clock hands size and hands drawing pointing at 2:50 hs. and 8:40 hs. were the variables that best discriminated AD, MCI and NC ( p<0.01). A stepwise regression analysis using as dependent variables (AD - NC ) and (MCI - NC) showed in both cases that the best predictor model was that formed by: planning strategies, clock hands pointing to 8:40, clock hands size, reading hours and set up clock hands following a formal numeric code (p<.001). CONCLUSIONS: Our proposed modifications in this test convert it in an appropriate tool for cognitive impairment screening because they do not only detect visuoconstructional abilities but also frontal planning, contents and processing of the hour memory code and transcodification of semantic memory processes. PMID- 15785785 TI - [Adverse effects of psychiatric drugs on glucose and lipids metabolism]. AB - Psychiatric drugs bring indeniable benefits for the treatment of psychiatric disorders. But their use can come along with a varying degree of side effects, some of them known for a long time, and some that are at present starting to focus attention. In this group we can find metabolic side effects, because of their possible relationship with the mortality of patients with severe psychiatric disorders. In this article, effects of psychiatric drugs on glucose and lipids metabolism are revised. PMID- 15785786 TI - [Liver disease induced by psychiatric drugs]. AB - The increase in the research, development and prescription of new psychotropic drugs used to treat mental illness, generated new and more frequent side effects. Drug-induced hepatotoxicity, in particular the idiosyncratic reactions, should be anticipated and diagnosed at an early stage due to their sudden and severe effects. We analyze the role of the liver in the drug metabolism, the different mechanisms of drug-induced hepatotoxicity of the main psychotropic drugs groups, and their clinical features. PMID- 15785787 TI - [Effects of psychiatric drugs on the fetus and newborn children. Consequences of the treatment of psychiatric disorders during pregnancy and lactation]. AB - The management of mental disorders during pregnancy and the postpartum period confronts the psychiatrist to a complex clinical situation. The cero risk utopia should be changed for a more realistic approach which considers the risks of having a conservative conduct (assuming the consequences of the untreated mental disorders) or making a psychopharmacological intervention knowing the expectable effects on the fetus and the offspring. The antidepressants and the conventional antipsychotics seem to be safe drugs in the first trimester of pregnancy. The benzodiazepines and the mood stabilizers are linked with the appearance of teratogenic malformations after the first trimester exposure. In the present article, the pharmacokinetic peculiarities of the pregnancy and the newborn and the general considerations of the psychopharmacological management of this kind of patients are described. At last, there is a review of the literature about the effects of the different groups of psychiatric drugs on the fetal growth, their passage to maternal milk and their impact on the newborn. PMID- 15785788 TI - [Endocrinologic adverse effects of psychotropic drugs]. AB - Psychotropic drugs affect the regulatory mechanisms of different neuroendocrine axis. This chapter reviews the interactions between psychotropic drugs and prolactin, the hypothalamic-pituitary-thyroid axis and the hypothalamic-pituitary gonadal axis. Hyperprolactinemia can cause galactorrhea, amenorrhea, sexual disfunction, impaired spermatogenesis and increased risk for osteoporosis and fractures. Atypical antipsychotics cause less hyperprolactinemia than conventional antipsychotics. Lithium has important effects on thyroid function. During lithium treatment, affectively ill patients show, in varying degrees and combinations, reduced levels of thyroid hormones and clinical evidence of subclinical hypothyroidism, overt hypothyroidism and goiter. Recent literature reports suggest that valproic acid, may be associated with polycistic ovarian syndrome. Until additional data is available, women starting valproate therapy should be warned about the possibility of endocrinology side effects. PMID- 15785789 TI - [Adverse effects of psychiatric drugs on sexual functions]. AB - Several groups of pharmacological agents have been reported to disrupt normal sexual function. Psychotropic medications, such as antidepressants and antipsychotics, have also been associated with sexual side effects. The procedure by which, the basic physiologic mechanisms of the normal sexual phases (libido, arousal, and orgasm) are disrupted by some psychotropics provide a framework to minimize sexual side effects when initiating and continuing treatment. Successful management of sexual complaints during treatment should begin with a systematic approach to determine the type of sexual dysfunction, potential contributing factors, and finally delineating strategies that should be tailored to the individual patient. This article provides guidelines for the assessment, management, and prevention of sexual side effects associated with antidepressant and antipsychotic treatment. PMID- 15785790 TI - [Tyramine and serotonin syndromes. Pharmacological, medical and legal remarks]. AB - The tyramine syndrome and the serotonin syndrome are a complex of signs and symptoms that are thought to be largely attributable to drug - drug interactions or drug - food interactions that enhances norepinephrine o serotonin activity. This article reviews: pharmacological basis of those syndromes; clinical features; forbidden foods, drug-drug interactions, and treatment options. Finally a set of legal recommendations are proposed to avoid liability litigations. PMID- 15785791 TI - [Social immunity]. PMID- 15785793 TI - Oxygen therapy, then and now. PMID- 15785794 TI - Use of spirometry and respiratory drugs in Manitobans over 35 years of age with obstructive lung diseases. AB - BACKGROUND: Previous data indicated that spirometry was underused in people with obstructive disease, especially those with chronic obstructive pulmonary disease (COPD). OBJECTIVE: To examine the use of respiratory drugs in patients with COPD and asthma, and to relate drug use to spirometry. METHODS: Manitoba Health maintains a database of physician services remunerated by fees that includes spirometry. The database contains the diagnosis and patient identifiers, as well as sex, date of birth and residential postal code. Similar identifiers are used in the provincial pharmacare program that records prescriptions dispensed at retail pharmacies. These databases were examined for the time period between 1996 to 2000, and people over 35 years of age diagnosed with asthma, COPD or both were identified. The frequency of spirometry in these patients and their use of respiratory drugs was determined. RESULTS: Spirometry and drug prescription frequencies increased with the number of physician visits (including those for bronchitis), but their patterns differed. Patients with asthma or asthma plus COPD had considerably higher rates of drug prescription and slightly higher spirometry rates than did those with COPD. Patients with asthma and asthma plus COPD who underwent spirometry were slightly more likely to receive drugs than those who did not undergo spirometry; this trend was more striking in patients with COPD. However, approximately 30% of patients with COPD who had five physician visits and who underwent spirometry did not receive drugs; this was true for approximately 10% of similar patients with asthma. Patients with asthma generally received beta-agonists and inhaled steroids; these agents were less commonly given to patients with COPD, who instead were given anticholinergics much more often than were asthmatics. Patients who were diagnosed with asthma plus COPD had beta-agonist and inhaled corticosteroid prescription rates similar to asthmatics, and anticholinergic prescription rates similar to patients with COPD. Theophylline and antileukotriene drugs were used less often than were inhaled agents. In patients with asthma, drugs were frequently discontinued, and during drug use, prescription refills were consistent with an intake of 30.9% of the prescribed doses. In patients with COPD, discontinuing drugs early was uncommon, and refills were consistent with the use of 54% of the prescribed amounts. The same was true of patients with both COPD and asthma. DISCUSSION: Drug prescription was considerably more common in patients labelled with asthma or COPD plus asthma than in patients with COPD. Spirometry was also less common in patients with COPD but had a distinct influence on the frequency of drug prescription. Patterns of drug prescription were predictable, and patterns of drug use indicated poor compliance, in agreement with other data. The results suggest that COPD symptoms may be discounted and patients systematically undertreated or the diagnosis could frequently be applied to people with trivial disease or both. PMID- 15785795 TI - Interpretation of transbronchial lung biopsies from lung transplant recipients: inter- and intraobserver agreement. AB - BACKGROUND: Transbronchial lung biopsy results are crucial for the management of lung transplant recipients. Little information is available regarding the reliability and reproducibility of the interpretation of transbronchial lung biopsies. OBJECTIVE: To examine the inter-reader variability between two lung pathologists with expertise in lung transplantation. METHODS: Fifty-nine transbronchial lung biopsy specimens were randomly selected. Active infection had been excluded in all cases. The original interpretations (as per the Lung Rejection Study Group) for acute rejection grade included 19 biopsies scored as A0 (none), 14 scored as A1 (minimal), 12 as A2 (mild), 11 as A3 (moderate) and three as A4 (severe). The pathologists worked independently without clinical information or knowledge of the original interpretation. The specimens were graded using the Lung Rejection Study Group criteria for acute rejection (grades A0 to A4), airway inflammation (grades B0 to B4) and bronchiolitis obliterans (C0 absent and C1 present). Between-reader agreement for each category was analyzed using a Kappa statistic. RESULTS: Because many transplant specialists initiate augmented immunosuppression with biopsy grades of A2 or higher, results for each reader were dichotomized as A0/A1 versus A2/A3/A4. Using this dichotomy, there was only moderate agreement (kappa 0.470, P < 0.001) between readers. For categories B and C, the results were dichotomized for the absence or presence of airway inflammation and bronchiolitis obliterans, respectively. The level of agreement between readers was fair for category B (kappa 0.333, P = 0.014) and poor for category C (kappa 0.166, P = 0.108). The intrareader agreement for acute rejection was substantial (kappa 0.795, P = 0.0001; kappa 0.676, P = 0.0001). CONCLUSIONS: Because the agreement between expert pathologists is only modest, optimum clinical decision-making requires that transbronchial lung biopsy results be used in an integrated clinical context. PMID- 15785796 TI - Cigarette smoking and asthma: a dangerous mix. AB - In Canada, 20% to 30% of the general population currently smoke. Smoking is as common in those suffering from asthma as it is in the general population. However, most studies on the pathophysiology of asthma and its response to treatment only include nonsmokers. Available data that examine the influence of smoking on clinical, functional and inflammatory characteristics of asthma, as well as the influence of smoking on the therapeutic response to corticosteroids, were reviewed. Active smoking is associated with an increased morbidity from asthma and impairs the response to inhaled corticosteroids. These observations emphasize the need for smoking cessation in patients with asthma and for reassessment of current treatment guidelines in this population. PMID- 15785797 TI - The introduction of oxygen for pneumonia as seen through the writings of two McGill University professors, William Osler and Jonathan Meakins. AB - Oxygen was identified at the end of the 18th century by three independent researchers. It was first used indiscriminately with other gases for treatment of pulmonary diseases by Thomas Beddoes. In the 19th century, the physiological properties of oxygen were identified by many researchers. In that same century, physicians used oxygen empirically for a variety of conditions. Osler, who wrote on pneumonia, appreciated that blood was "imperfectly oxidised" (sic) in pneumonia, but concluded that the toxicity of oxygen more than outweighed its possible benefits. Meakins applied the lessons he learned from studying the hypoxemia that resulted from poison gas in World War I to pneumonia. He confirmed that patients with severe pneumonia were hypoxemic and that many of their symptoms appeared to be relieved by inhalation of oxygen. Oxygen then became the standard therapy for pneumonia. PMID- 15785798 TI - Laser therapy for an obstructing primary tracheal lymphoma in a patient with AIDS. AB - A patient with AIDS presented with progressive dyspnea leading to respiratory failure with near complete airway obstruction due to primary tracheal lymphoma. Laser therapy was used locally to debulk the tumour, which facilitated extubation and led to definitive treatment with chemotherapy. Alternatives for local airway control are discussed. PMID- 15785799 TI - Chemical biology of protein lipidation: semi-synthesis and structure elucidation of prenylated RabGTPases. AB - Rab/Ypt guanosine triphosphatases (GTPases) represent a family of key membrane traffic regulators in eukaryotic cells. For their function Rab/Ypt proteins require double modification with two covalently bound geranylgeranyl lipid moieties at the C-terminus. Generally, prenylated proteins are very difficult to obtain by recombinant or enzymatic methods. We generated prenylated RabGTPases using a combination of chemical synthesis and protein engineering. This semi synthesis depends largely on the availability of functionalized prenylated peptides corresponding to the proteins' native structure or modifications. We developed solution phase and solid phase strategies for the generation of peptides corresponding to the prenylated C-terminus of Rab7 GTPase in preparative amounts enabling us to crystallize the mono-prenylated Ypt1:RabGDI complex. The structure of the complex provides a structural basis for the ability of RabGDI to inhibit the release of nucleotide by Rab proteins and a molecular basis for understanding a RabGDI mutant that causes mental retardation in humans. PMID- 15785800 TI - Synthesis of technomimetic molecules: towards rotation control in single molecular machines and motors. AB - Technomimetic molecules are molecules designed to imitate macroscopic objects at the molecular level, also transposing the motions that these objects are able to undergo. This article focuses on technomimetic molecules with rotary motions, including gears, wheelbarrows and motors. Following the bottom-up approach the synthesis of technomimetic molecules grants access to the study of mechanical properties at the molecular level. These molecules are designed to operate as single molecules on surfaces under the control of the tip of a scanning tunneling microscope or atomic force microscope. PMID- 15785801 TI - Synthesis and crystal structure of a salen-type copper(II) complex derived from 3,5'-O-dimethyl-2',3'-diamino-2',3'-dideoxy-beta-D-xylo-uridine. AB - We report the synthesis of the first 2',3'-diamino substituted uridine derivative with xylo-configuration, its use as starting material for the synthesis of salen type ligands and the first structurally characterised copper(II) complex thereof. PMID- 15785802 TI - Bassianolone: an antimicrobial precursor of cephalosporolides E and F from the entomoparasitic fungus Beauveria bassiana. AB - We have established the chemical structure of (+)-bassianolone (3), the antimicrobial compound precursor of cephalosporolides E and F, and that of the furan metabolite 4 from the entomopathogenic fungus Beauveria bassiana. PMID- 15785804 TI - Prediction of vapour pressure using descriptors derived from molecular dynamics. AB - Vapour pressures of organic materials can be predicted to high levels of accuracy using cohesive energies and solubility parameters derived from molecular dynamics simulations that use good forcefields. It is found that over 90% of the correlation with vapour pressure is accounted for by a single cross term involving the product of either the molecular weight or molar volume of a molecule and its cohesive energy density. PMID- 15785803 TI - On the origin of the DNA sequence selectivity of the azinomycins. AB - Simplified synthetic azinomycins preferentially induce in vitro DNA interstrand cross-links at the same 5'-d(GCC)-3' site as the natural products revealing that non-covalent interactions are relatively unimportant in defining sequence specificity. PMID- 15785805 TI - Theoretical study of aza-polycyclic aromatic hydrocarbons (aza-PAHs), modelling carbocations from oxidized metabolites and their covalent adducts with representative nucleophiles. AB - Protonation of the epoxides, diol epoxides, and dihydrodiols of benzo[h]quinoline (BhQ), benzo[f]quinoline (BfQ), phenanthrene (Phe), benzo[c]phenanthridine (BcPhen), and chrysene (Chry) were studied by DFT at the B3LYP/6-31G* level, and selected cases were calculated with the 6-31+G* diffuse-function augmented basis set for comparison purposes. Bay-region carbocations were formed from O protonated epoxides via a barrierless processes. Relative carbocation stabilities were determined in the gas phase and with water as solvent (PCM method). The presence of a heteroatom changes the regioselectivity of epoxide ring opening, in some cases favoring non-bay-region carbocations. The epoxide ring opening mode is also greatly influenced by N-protonation. The dications resulting from initial N protonation followed by epoxide protonation were also studied by DFT. Charge delocalization modes in the resulting mono- and dications were derived by GIAO NMR (based on Delta delta13C values) and via the NPA-derived changes in charges. Relative aromaticity in different rings in the arenium ions was gauged by NICS. In representative cases, the covalent adducts (syn and anti) formed by reaction of the benzylic carbocations derived from diol epoxides and dihydrodiols with methoxide and methanethiolate anions were studied. Relative energies (in the gas phase and with water as solvent) and geometries of the adducts formed by quenching of the carbocations derived from BhQ and Phe-epoxides with guanine via the exocyclic amino group and via the N-7 were also investigated computationally. Although aqueous phase calculations change the energy for the addition reactions because of greater stabilization of the reactants, relative reactivity trends remain the same. The data are discussed, taking into account the available experimental results concerning the biological activity of these compounds. PMID- 15785806 TI - Advanced approaches for the characterization of a de novo designed antiparallel coiled coil peptide. AB - We report here an advanced approach for the characterization of the folding pattern of a de novo designed antiparallel coiled coil peptide by high-resolution methods. Incorporation of two fluorescence labels at the C- and N-terminus of the peptide chain as well as modification of two hydrophobic core positions by Phe/[15N,13C]Leu enable the study of the folding characteristics and of distinct amino acid side chain interactions by fluorescence resonance energy transfer (FRET) and NMR spectroscopy. Results of both experiments reveal the antiparallel alignment of the helices and thus prove the design concept. This finding is also supported by molecular dynamics simulations. Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS) in combination with NMR experiments was used for verification of the oligomerization equilibria of the coiled coil peptide. PMID- 15785807 TI - New aminocyclitols as modulators of glucosylceramide metabolism. AB - A series of 13 aminocyclitol derivatives belonging to two different families is described. Their configuration is governed by the regio- and stereocontrolled epoxide opening of a suitably protected conduritol-B epoxide. Studies on several glycosyl processing enzymes indicate that some of them are good inhibitors of glucosylceramide hydrolase. A rationale to account for preliminary structure activity relationships is provided. PMID- 15785808 TI - Stereoselective electrochemical carboxylation: 2-phenylsuccinates from chiral cinnamic acid derivatives. AB - Chiral 2-phenyl succinic ester derivatives have been obtained under mild conditions, in short times and with satisfactory yields by electrochemical reduction of chiral cinnamic acid derivatives under a CO2 atmosphere. When 4R (diphenylmethyl)-oxazolidin-2-one was used as a chiral auxiliary the two diastereoisomers could be easily separated by flash chromatography and the R isomer was obtained as major product. PMID- 15785809 TI - Piperazine additions to C60--a facile approach to fullerene substitution. AB - A range of fullerene monoadducts can be generated via the photochemical reaction of piperazine derivatives with C60. Addend functionality can also be efficiently incorporated by transformation of the hydroxyl-substituted adduct prepared in this fashion. Reaction yields and process simplicity compete with current standard procedures for fullerene mono-functionalisation. PMID- 15785810 TI - Solution and solid state structure and tautomerism of azo coupled enaminone derivatives of benzoylacetone. AB - The reaction of 4-substituted benzenediazonium tetrafluoroborates with 3-amino-1 phenylbut-2-en-1-one, 4-amino-4-phenylbut-3-en-2-one and their N-aryl derivatives 1a-1g has been used to prepare the respective azo coupling products i.e. compounds 2-5 from enaminone 1a, compounds 6-9 from enaminone 1c, compound 10 from enaminone 1d, compound 11 from enaminone 1e, compounds 12, 13 from enaminone 1f, compounds 14, 15 from enaminone 1b and compound 16 from enaminone 1g. Tautomerism of the azo coupling products prepared has been investigated in CDCl3 solutions by means of 1H, 13C and 15N NMR spectra. Crystal structures of selected products have also been investigated by means of X-ray diffraction. PMID- 15785811 TI - Structure elucidation of cyasterone stereoisomers isolated from Cyathula officinalis. AB - Chemical investigation of ecdysteroidal constituents of the roots and stems of Cyathula officinalis led to the isolation of two cyasterone stereoisomers, 2 and 3, together with the known cyasterone 1. The structures of compounds 2 and 3 were determined to be 28-epi-cyasterone and 25-epi-28-epi-cyasterone, respectively, by means of spectroscopic analysis. X-Ray structures of 1 and 2 confirmed the 24S,25S,28R configuration for 1 and 24S,25S,28S for 2. PMID- 15785812 TI - An improved solid-phase methodology for the synthesis of putative hexa- and heptapeptide intermediates in vancomycin biosynthesis. AB - The biosynthesis of the vancomycin aglycone involves three oxidative phenol coupling reactions, each catalyzed by a discrete cytochrome P450-like enzyme. Studies on the mechanism and specificity of the enzyme (called OxyB) catalyzing the first coupling, require access to suitable linear peptide precursors, each conjugated as a thioester to a peptide carrier domain of the vancomycin non ribosomal peptide synthetase. An efficient route to representative free linear peptides is described here. The method makes use of Alloc-chemistry during solid phase assembly of the peptide backbone, but importantly and in contrast to earlier efforts, largely avoids the use of amino acid side chain protecting groups. In this way, the target linear peptides can be released directly from the solid support under very mild conditions. PMID- 15785813 TI - Kinetics and mechanism of the aminolysis of aryl thiocarbamates: effects of the non-leaving group. AB - The kinetics of the aminolysis of aryl thiocarbamates [ATC: H2NC(=O)SC6H4Z] with benzylamines (XC6H4CH2NH2) in acetonitrile at 10.0 degrees C have been studied. The rate order with variation of the non-leaving amino group, RNH, in RNHC(=O)SC6H4Z is NH2 < PhNH < EtNH indicating that the polar (sigma*) and steric (E(s)) effects of the RNH group are insignificant, and the strength of push to expel the leaving group in the tetrahedral transition state is the sole, important effect. The strong push provided by the NH2 group, the negative rhoXZ( 0.38) value, the size of betaZ(-0.54), and failure of the reactivity-selectivity principle are all consistent with the concerted mechanism. The kinetic isotope effects involving deuterated amine nucleophiles (XC6H4CH2ND2) are normal (k(H)/k(D)approximately 1.40-1.73) suggesting a hydrogen-bonded cyclic transition state. PMID- 15785814 TI - Synthesis of 5'-methylenearisteromycin and its 2-fluoro derivative with potent antimalarial activity due to inhibition of the parasite S-adenosylhomocysteine hydrolase. AB - 5'-methylenearisteromycin 5 and its 2-fluoro derivative 6, which were designed as antimalarial agents because of their AdoHcy hydrolase inhibition, were synthesized from D-ribose, using a stereoselective intramolecular radical cyclization as the key step to construct the carbocyclic structure. These compounds were evaluated as AdoHcy hydrolase inhibitors with the recombinant human and malarial parasite enzymes. Although 5 and 6 were both potent inhibitors of the malarial parasite AdoHcy hydrolase, the 2-fluoro derivative 6 proved to be superior due to its lower inhibitory effect on the human enzyme. In addition, 6 was identified as a potent antimalarial agent using an in vitro assay system with Plasmodium falciparum. PMID- 15785815 TI - Chiral oxime ethers in asymmetric synthesis. O-(1 Phenylbutyl)benzyloxyacetaldoxime, a versatile reagent for the asymmetric synthesis of protected 1,2-aminoalcohols, alpha-amino acid derivatives, and 2 hydroxymethyl nitrogen heterocycles including iminosugars. AB - Addition of a range of organolithium and Grignard reagents to (E)-O-(1 phenylbutyl)benzyloxyacetaldoxime 1 in the presence of boron trifluoride diethyl etherate is highly diastereoselective. The resulting hydroxylamines undergo N-O bond cleavage upon treatment with zinc-acetic acid or molybdenum hexacarbonyl to give, after N-protection, protected 1,2-aminoalcohols 3 in high enantiomeric purity. Debenzylation of 3a and 3d gave N-Boc (R)-alaninol and (S)-phenylalaninol respectively. The hydroxylamines 2 also serve as alpha-amino acid precursors, 2i being converted into N-formyl-(R)-alaninyl-(S)-(4-bromo)phenylalanine ester 7, the N-terminal dipeptide of a natural depsipeptide. The versatility of the 1,2 aminoalcohol derivatives was further illustrated by their conversion into 5-, 6- and 7-membered 2-hydroxymethyl nitrogen heterocycles 15-19 in high enantiomeric excess by a ring-closing metathesis reaction. Further reaction of the dihydropyrrole 15 gave the iminosugar 1,4-dideoxy-1,4-imino-D-ribitol. PMID- 15785816 TI - Synthesis of multisubstituted 1,3-butadienes using the ruthenium-catalysed double addition of trimethylsilyldiazomethane to alkynylboronates. AB - The ruthenium-catalysed double addition of trimethylsilyldiazomethane to alkynes developed by Dixneuf and co-workers was applied to the synthesis of 2-alkyl- or 2 aryl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,4-bis(trimethylsilyl)-1,3 butadienes by use of alkynylboronates instead of alkynes. Di- and tetrasubstituted 1,3-butadienes were prepared from a 2-boryl-1,4-disilyl-1,3 butadiene, using the Suzuki-Miyaura coupling reaction, iodolysis of the alkenylsilane moieties with N-iodosuccinimide and hydrolysis of the carbon silicon bonds with trifluoroacetic acid. The same compound was converted also to a bicyclic compound, a trisubstituted 1,3-butadiene and a dienone through the Diels-Alder reaction, oxidation of the alkenylboronate moiety and the Mukaiyama aldol reaction. PMID- 15785817 TI - Nitroxides with two pK values--useful spin probes for pH monitoring within a broad range. AB - A series of 4-dialkylamino-2,5-dihydroimidazole nitroxides with pyridine-4-yl, 4 dimethylaminophenyl or 4-hydroxyphenyl groups in position 2 of the imidazole ring were prepared using the reaction of RMgBr with corresponding 5-dialkylamino-4,4 dimethyl-4H-imidazole 3-oxides. The EPR spectra of the nitroxides were shown to be pH-sensitive due to consecutive protonation of the amidino moiety and the basic group(s) at position 2 of the imidazole ring. The 5,5-dimethyl-4 (dimethylamino)-2-ethyl-2-pyridine-4-yl-2,5-dihydro-1H-imidazol-1-oxyl showed a monotonic increase in the isotropic nitrogen hyperfine (hfi) coupling constant alpha(N) of 1 .4 G over a pH range from 2 to 6.5. Such a broad range of pH sensitivity could be useful for many biophysical and biomedical applications, including pH-monitoring in the stomach. PMID- 15785818 TI - New fluorescent probes reveal that flippase-mediated flip-flop of phosphatidylinositol across the endoplasmic reticulum membrane does not depend on the stereochemistry of the lipid. AB - Glycerophospholipid flip-flop across biogenic membranes such as the endoplasmic reticulum (ER) is a fundamental feature of membrane biogenesis. Flip-flop requires the activity of specific membrane proteins called flippases. These proteins have yet to be identified in biogenic membranes and the molecular basis of their action is unknown. It is generally believed that flippase-facilitated glycerophospholipid flip-flop across the ER is governed by the stereochemistry of the glycerolipid, but this important issue has not been resolved. Here we investigate whether the ER flippase stereochemically recognizes the glycerophospholipids that it transports. To address this question we selected phosphatidylinositol (PI), a biologically important molecule with chiral centres in both its myo-inositol headgroup and its glycerol-lipid tail. The flip-flop of PI across the ER has not been previously reported. We synthesized fluorescence labeled forms of all four diastereoisomers of PI and evaluated their flipping in rat liver ER vesicles, as well as in flippase-containing proteoliposomes reconstituted from a detergent extract of ER. Our results show that the flippase is able to translocate all four PI isomers and that both glycerol isomers of PI flip-flop across the ER membrane at rates similar to that measured for fluorescence-labeled phosphatidylcholine. Our data have important implications for recent hypotheses concerning the evolution of distinct homochiral glycerophospholipid membranes during the speciation of archaea and bacteria/eukarya from a common cellular ancestor. PMID- 15785819 TI - Cyclic beta-amino acid derivatives: synthesis via lithium amide promoted tandem asymmetric conjugate addition-cyclisation reactions. AB - The product distribution upon conjugate addition of homochiral lithium N-benzyl-N alpha-methylbenzylamide to dimethyl-(E,E)-nona-2,7-dienedioate can be controlled to give either the cyclic 1,2-anti-1,6-anti-beta-amino ester (derived from conjugate addition and intramolecular enolate cyclisation) or the acyclic bis beta-amino ester derivative (derived from double conjugate addition) in high de. The introduction of a protected nitrogen functionality into the diester skeleton facilitates, after conjugate addition and intramolecular enolate cyclisation, the asymmetric construction of piperidines in high de; variation in the N-protecting group indicates that the highest stereoselectivity is observed with alpha branched N-substituents. Tandem conjugate addition-aldol reactions can also be achieved stereoselectively, with lithium amide conjugate addition to epsilon- and zeta-oxo-alpha,beta-unsaturated esters giving the corresponding five and six membered cyclic beta-amino esters in high de. N-deprotection by hydrogenolysis of the products arising from these reactions furnishes a range of polyfunctionalised transpentacin and transhexacin derivatives in high de and ee. PMID- 15785820 TI - On the Diels-Alder reactions of pentadienyl maleates and citraconates. AB - Reactions between conjugated dienols and maleic anhydride provide either cis fused or trans-fused bicyclic products as major products, depending upon how the reaction is carried out. Simply mixing the two reactants together generally leads to cis-fused lactone acids in thermal reactions which proceed viaintermolecular Diels-Alder reaction followed by intramolecular esterification. Pre-forming the maleate half ester derivative followed by heating affords predominantly trans fused lactone acids in good yields by way of an intramolecular Diels-Alder (IMDA) reaction. Sorbyl citraconate half esters undergo a rapid thermolytic fragmentation in refluxing toluene to form the dienol and citraconic anhydride. The resulting diene-dienophile pair undergo an intermolecular cycloaddition followed by a rapid intramolecular esterification to give cis-fused bicyclic lactone acids as major products. The IMDA reaction of citraconic half esters is sufficiently rapid in DMSO to dominate over fragmentation: the exo-cycloadduct is formed almost exclusively. Nine literature reports of endo-selective IMDA reactions of triene acids are erroneous; the cycloadditions proceed in an intermolecular manner. PMID- 15785821 TI - Ring fission of chiral cyclic acetals plus intramolecular [4 + 2] cycloaddition: a sequential access to medium-size lactones. Application to the synthesis of carbasugars. AB - A set of alpha,beta-unsaturated cyclic acetals were reacted with t-butyllithium in THF. A conjugate elimination took place, triggering a ketal ring cleavage when two eq. of TMEDA were added to the medium. The lithium alcoholate thus obtained could be trapped in situ by 2,2,2-trifluoroethyl acrylate. The resulting acryloyl (1Z,3E)-dienyl ether was then submitted to an intramolecular Diels-Alder cycloaddition under high pressure (12 kbar, 50 degrees C). Depending on the structure of the linkage between the diene and the acrylate, the expected ("fused") lactones were obtained with total endo- or exo-selectivities and high to complete diastereoselectivities. A remarkable inversion of selectivity, from endo to exo, with respect to the stereochemical elements of the tether could be observed in these cases. A five step diastereoselective transformation of two of the resulting nine-membered ring lactones into modified carbasugars was finally achieved in 23% overall yield. PMID- 15785822 TI - Xanthenol clathrates: structure, thermal stability, guest exchange and kinetics of desolvation. AB - A series of clathrates comprising the xanthenol host, 9-(4-methoxyphenyl)-9H xanthen-9-ol, with a variety of aromatic guests displays similar structures in the space group P(-1). We have elucidated the structures of the inclusion compounds H x 1/2G, where H is 9-(4-methoxyphenyl)-9H-xanthen-9-ol and G is benzene, o-, m- and p-xylene. The structures are isostructural with respect to the host and display consistent (Host)-OH...O-(Host) hydrogen bonding. The guests lie on a centre of inversion and with the exception of the symmetrical guests, benzene and p-xylene, are disordered. An interesting case arises with m-xylene, which is ordered at low temperature (113 K) with both the host and guest molecules in general positions. At a higher temperature (283 K) the inclusion compound with m-xylene fits the series. We have correlated the structures with their thermal stabilities, guest exchange and kinetics of desolvation. PMID- 15785823 TI - Complete ozonolysis of alkyl substituted ethenes at -60 degrees C: distributions of ozonide and oligomeric products. AB - The distribution of ozonide and oligomeric structures formed on complete ozonolysis of alkenes in a non-participating solvent at -60 degrees C is governed by the alkyl substitution around the carbon-carbon double bond. The ozonolysis of a 1,1-alkyl substituted ethene generally favours the formation of an ozonide (a 1,2,4-trioxolane). Whereas the ozonolysis of a 1,1,2-alkyl substituted ethene also produces ozonide, a considerable amount of the ozonised products are oligomeric in nature. For example, the ozonolysis of 3-methylpent-2-ene in solution to high conversion in pentane yields oligomers with structural units derived from the fragmentation products of the primary ozonide (a 1,2,3 trioxolane) which are namely butanone carbonyl oxide and acetaldehyde; these can be characterised by electrospray ionisation mass spectroscopy (ESI-MS) under soft ionisation conditions. The predominant oligomers formed are rich in carbonyl oxide units (80 + mol%) and are cyclic in nature. A small proportion of the oligomers formed are open chain compounds with end groups that suggest that chain termination is brought about either by water or by hydrogen peroxide. Residual water in the solvent will react with the carbonyl oxides to produce 2-methoxybut 2-yl hydroperoxide, which we propose readily decomposes generating hydrogen peroxide. A significant yield of oligomers also is obtained from the ozonolysis of a 1,2-alkyl substituted ethene. The ozonolysis of trans-hex-2-ene in pentane yields oligomers containing up to four structural units and are predicted to be mainly cyclic. PMID- 15785824 TI - Comments on the paper "c-erbB-2 expression and nuclear pleomorphism in canine mammary tumors". PMID- 15785826 TI - Corticosteroids in acute respiratory distress syndrome. AB - Improving the course and outcome of patients with acute respiratory distress syndrome presents a challenge. By understanding the immune status of a patient, physicians can consider manipulating proinflammatory systems more rationally. In this context, corticosteroids could be a therapeutic tool in the armamentarium against acute respiratory distress syndrome. Corticosteroid therapy has been studied in three situations: prevention in high-risk patients, early treatment with high-dose, short-course therapy, and prolonged therapy in unresolving cases. There are differences between the corticosteroid trials of the past and recent trials: today, treatment starts 2-10 days after disease onset in patients that failed to improve; in the past, the corticosteroid doses employed were 5-140 times higher than those used now. Additionally, in the past treatment consisted of administering one to four doses every 6 h (methylprednisolone, 30 mg/kg) versus prolonging treatment as long as necessary in the new trials (2 mg kg(-1) day(-1) every 6 h). The variable response to corticosteroid treatment could be attributed to the heterogeneous biochemical and molecular mechanisms activated in response to different initial insults. Numerous factors need to be taken into account when corticosteroids are used to treat acute respiratory distress syndrome: the specificity of inhibition, the duration and degree of inhibition, and the timing of inhibition. The major continuing problem is when to administer corticosteroids and how to monitor their use. The inflammatory mechanisms are continuous and cyclic, sometimes causing deterioration or improvement of lung function. This article reviews the mechanisms of action of corticosteroids and the results of experimental and clinical studies regarding the use of corticosteroids in acute respiratory distress syndrome. PMID- 15785828 TI - Increased production of tumor necrosis factor-alpha in whole blood cultures from children with primary malnutrition. AB - Because low tumor necrosis factor-alpha (TNF-alpha) production has been reported in malnourished children, in contrast with high production of TNF-alpha in experimental protein-energy malnutrition, we reevaluated the production of TNF alpha in whole blood cultures from children with primary malnutrition free from infection, and in healthy sex- and age-matched controls. Mononuclear cells in blood diluted 1:5 in endotoxin-free medium released TNF-alpha for 24 h. Spontaneously released TNF-alpha levels (mean +/- SEM), as measured by enzyme immunoassay in the supernatants of unstimulated 24-h cultures, were 10,941 +/- 2,591 pg/ml in children with malnutrition (N = 11) and 533 +/- 267 pg/ml in controls (N = 18) (P < 0.0001). TNF-alpha production was increased by stimulation with lipopolysaccharide (LPS), with maximal production of 67,341 +/- 16,580 pg/ml TNF-alpha in malnourished children and 25,198 +/- 2,493 pg/ml in controls (P = 0.002). In control subjects, LPS dose-dependently induced TNF-alpha production, with maximal responses obtained at 2000 ng/ml. In contrast, malnourished patients produced significantly more TNF-alpha with 0.02-200 ng/ml LPS, responded maximally at a 10-fold lower LPS concentration (200 ng/ml), and presented high dose inhibition at 2000 ng/ml. TNF-alpha production a) was significantly influenced by LPS concentration in control subjects, but not in malnourished children, who responded strongly to very low LPS concentrations, and b) presented a significant, negative correlation (r = -0.703, P = 0.023) between spontaneous release and the LPS concentration that elicited maximal responses in malnourished patients. These findings indicate that malnourished children are not deficient in TNF-alpha production, and suggest that their cells are primed for increased TNF alpha production. PMID- 15785827 TI - RANK, RANKL and osteoprotegerin in arthritic bone loss. AB - Rheumatoid arthritis is characterized by the presence of inflammatory synovitis and destruction of joint cartilage and bone. Tissue proteinases released by synovia, chondrocytes and pannus can cause cartilage destruction and cytokine activated osteoclasts have been implicated in bone erosions. Rheumatoid arthritis synovial tissues produce a variety of cytokines and growth factors that induce monocyte differentiation to osteoclasts and their proliferation, activation and longer survival in tissues. More recently, a major role in bone erosion has been attributed to the receptor activator of nuclear factor kappa B ligand (RANKL) released by activated lymphocytes and osteoblasts. In fact, osteoclasts are markedly activated after RANKL binding to the cognate RANK expressed on the surface of these cells. RANKL expression can be upregulated by bone-resorbing factors such as glucocorticoids, vitamin D3, interleukin 1 (IL-1), IL-6, IL-11, IL-17, tumor necrosis factor-alpha, prostaglandin E2, or parathyroid hormone related peptide. Supporting this idea, inhibition of RANKL by osteoprotegerin, a natural soluble RANKL receptor, prevents bone loss in experimental models. Tumor growth factor-beta released from bone during active bone resorption has been suggested as one feedback mechanism for upregulating osteoprotegerin and estrogen can increase its production on osteoblasts. Modulation of these systems provides the opportunity to inhibit bone loss and deformity in chronic arthritis. PMID- 15785829 TI - One-year follow-up of the effects of sildenafil on pulmonary arterial hypertension and veno-occlusive disease. AB - We hypothesized that chronic oral administration of the phosphodiesterase-5 inhibitor sildenafil could improve the exercise capacity and pulmonary hemodynamics in patients with pulmonary arterial hypertension (PAH) on the basis of previous short-term studies. We tested this hypothesis in 14 subjects with PAH, including seven patients with the idiopathic form and seven patients with atrial septal defects, but no other congenital heart abnormalities. Patients were subjected to a 6-min walk test and dyspnea was graded according to the Borg scale. Pulmonary flow and pressures were measured by Doppler echocardiography. Patients were given sildenafil, 75 mg orally three times a day, and followed up for 1 year. Sildenafil therapy resulted in the following changes: increase in the 6-min walk distance from a median value of 387 m (range 0 to 484 m) to 462 m (range 408 to 588 m; P < 0.01), improvement of the Borg dyspnea score from 4.0 (median value) to 3.0 (P < 0.01), and increased pulmonary flow (velocity-time integral) from a median value of 0.12 (range 0.08 to 0.25) to 0.23 (range 0.11 to 0.40; P < 0.01) with no changes in pulmonary pressures. In one patient with pulmonary veno-occlusive disease diagnosed by a lung biopsy, sildenafil had a better effect on the pulmonary wedge pressure than inhaled nitric oxide (15 and 29 mmHg, respectively, acute test). He walked 112 m at baseline and 408 m at one year. One patient died at 11 months of treatment. No other relevant events occurred. Thus, chronic administration of sildenafil improves the physical capacity of PAH patients and may be beneficial in selected cases of veno occlusive disease. PMID- 15785830 TI - Bronchial hyperresponsiveness and analysis of induced sputum cells in Crohn's disease. AB - With the aim of investigating the presence of latent inflammatory process in the lungs of patients with Crohn's disease, 15 patients with Crohn's disease were evaluated by spirometry, the methacholine challenge test, induced sputum, and skin tests for inhaled antigens. Serum IgE, erythrocyte sedimentation rate and hematocrit were also determined. The patients were compared with 20 healthy controls by the Mann-Whitney and Fisher exact tests. Their respiratory physical examination was normal. None had a personal or family history of clinical atopy. None had a previous history of pulmonary disease, smoking or toxic bronchopulmonary exposure. None had sinusitis, migraine, diabetes mellitus, or cardiac failure. Four (26.6%) of the patients with Crohn's disease had a positive methacholine challenge test whereas none of the 20 controls had a positive methacholine test (P = 0.026, Fisher exact test). Patients with Crohn's disease had a higher level of lymphocytes in induced sputum than controls (mean 14.59%, range 3.2-50 vs 5.46%, 0-26.92%, respectively; P = 0.011, Mann-Whitney test). Patients with Crohn's disease and a positive methacholine challenge test had an even higher percentage of lymphocytes in induced sputum compared with patients with Crohn's disease and a negative methacholine test (mean 24.88%, range 12.87 50 vs 10.48%, 3.2-21.69%; P = 0.047, Mann-Whitney test). The simultaneous findings of bronchopulmonary lymphocytosis and bronchial hyperresponsiveness in patients with Crohn's disease were not reported up to now. These results suggest that patients with Crohn's disease present a subclinical inflammatory process despite the absence of pulmonary symptoms. PMID- 15785831 TI - Baroreflex function in conscious rats submitted to iron overload. AB - Our hypothesis is that iron accumulated in tissue, rather than in serum, may compromise cardiovascular control. Male Fischer 344 rats weighing 180 to 220 g were divided into 2 groups. In the serum iron overload group (SIO, N = 12), 20 mg elemental iron was injected ip daily for 7 days. In the tissue iron overload group (TIO, N = 19), a smaller amount of elemental iron was injected (10 mg, daily) for 5 days followed by a resting period of 7 days. Reflex heart rate responses were elicited by iv injections of either phenylephrine (0.5 to 5.0 microg/kg) or sodium nitroprusside (1.0 to 10.0 microg/kg). Baroreflex curves were determined and fitted to sigmoidal equations and the baroreflex gain coefficient was evaluated. To evaluate the role of other than a direct effect of iron on tissue, acute treatment with the iron chelator deferoxamine (20 mg/kg, iv) was performed on the TIO group and the baroreflex was re-evaluated. At the end of the experiments, evaluation of iron levels in serum confirmed a pronounced overload for the SIO group (30-fold), in contrast to the TIO group (2-fold). Tissue levels of iron, however, were higher in the TIO group. The SIO protocol did not produce significant alterations in the baroreflex curve response, while the TIO protocol produced a nearly 2-fold increase in baroreflex gain (-4.34 +/- 0.74 and -7.93 +/- 1.08 bpm/mmHg, respectively). The TIO protocol animals treated with deferoxamine returned to sham levels of baroreflex gain (-3.7 +/- 0.3 sham vs -3.6 +/- 0.2 bpm/mmHg) 30 min after the injection. Our results indicate an effect of tissue iron overload on the enhancement of baroreflex sensitivity. PMID- 15785832 TI - Twenty-four-hour esophageal pH monitoring in children and adolescents with chronic and/or recurrent rhinosinusitis. AB - Gastroesophageal reflux (GER) disorder was studied in children and adolescents with chronic and/or recurrent rhinosinusitis not associated with bronchial asthma. Ten children with a clinical and radiological diagnosis of chronic and/or recurrent rhinosinusitis, consecutively attended at the Pediatric Otolaryngology Outpatient Clinic, Federal University of Sao Paulo, were evaluated. Prolonged esophageal pH monitoring was used to investigate GER disorder. The mean age of the ten patients evaluated (eight males) was 7.4 +/- 2.4 years. Two patients presented vomiting as a clinical manifestation and one patient presented retrosternal pain with a burning sensation. Twenty-four-hour esophageal pH monitoring was performed using the Sandhill apparatus. An antimony probe electrode was placed in the lower third of the esophagus, confirmed by fluoroscopy and later by a chest X-ray. The parameters analyzed by esophageal pH monitoring included: total percent time of the presence of acid esophageal pH, i.e., pH below 4 (<4.2%); total number of acid episodes (<50 episodes); number of reflux episodes longer than 5 min (3 or less), and duration of the longest reflux episode (<9.2 min). One patient (1/10, 10%) presented a 24-h esophageal pH profile compatible with GER disorder. This data suggest that an association between chronic rhinosinusitis not associated with bronchial asthma and GER disorder may exist in children and adolescents, especially in those with compatible GER disorder symptoms. In these cases, 24-h esophageal pH monitoring should be performed before indicating surgery, since the present data suggest that 10% of chronic rhinosinusitis surgeries can be eliminated. PMID- 15785833 TI - Prevalence of retinopathy in Caucasian type 2 diabetic patients from the South of Brazil and relationship with clinical and metabolic factors. AB - Diabetic retinopathy (DR) is a sight-threatening chronic complication of diabetes mellitus and is the leading cause of acquired blindness in adults. In this cross sectional study, we investigated the prevalence of and the factors associated with DR in an analysis of 210 consecutive and unrelated Brazilian Caucasians with type 2 diabetes mellitus. Retinopathy was evaluated by ophthalmoscopy and/or biomicroscopy through dilated pupils. The relationship between clinical and metabolic variables and the presence of DR was assessed by logistic regression analysis. DR was detected in 99 of the 210 patients (47%). In the univariate logistic regression analyses, male sex, duration of diabetes, body mass index, glycated hemoglobin, C-peptide, LDL cholesterol, smoking, and albumin excretion rate were found to be associated with the presence of DR. However, the multiple logistic regression analysis showed that only duration of diabetes (odds ratio (OR) = 1.15, 95% CI = 1.09-1.22; P < 0.001), glycated hemoglobin (OR = 1.21, 95% CI = 1.01-1.46; P = 0.047) and albumin excretion rate > 100 microg/min (OR = 12.72, 95% CI = 3.89-41.56; P < 0.001) were independently associated with DR. Although DR was found to be frequent among Brazilian type 2 diabetic patients, its prevalence was within the range observed in other Caucasian populations. Our findings emphasize the need for good glycemic control in order to prevent or delay the onset of DR, since the most well-known risk factors for the development of this complication in type 2 diabetes mellitus, such as duration of diabetes, glycated hemoglobin and albumin excretion rate were independently related to DR. PMID- 15785834 TI - Effect of air pollution on pediatric respiratory emergency room visits and hospital admissions. AB - In order to assess the effect of air pollution on pediatric respiratory morbidity, we carried out a time series study using daily levels of PM10, SO2, NO2, ozone, and CO and daily numbers of pediatric respiratory emergency room visits and hospital admissions at the Children's Institute of the University of Sao Paulo Medical School, from August 1996 to August 1997. In this period there were 43,635 hospital emergency room visits, 4534 of which were due to lower respiratory tract disease. The total number of hospital admissions was 6785, 1021 of which were due to lower respiratory tract infectious and/or obstructive diseases. The three health end-points under investigation were the daily number of emergency room visits due to lower respiratory tract diseases, hospital admissions due to pneumonia, and hospital admissions due to asthma or bronchiolitis. Generalized additive Poisson regression models were fitted, controlling for smooth functions of time, temperature and humidity, and an indicator of weekdays. NO2 was positively associated with all outcomes. Interquartile range increases (65.04 microg/m3) in NO2 moving averages were associated with an 18.4% increase (95% confidence interval, 95% CI = 12.5-24.3) in emergency room visits due to lower respiratory tract diseases (4-day moving average), a 17.6% increase (95% CI = 3.3-32.7) in hospital admissions due to pneumonia or bronchopneumonia (3-day moving average), and a 31.4% increase (95% CI = 7.2-55.7) in hospital admissions due to asthma or bronchiolitis (2-day moving average). The study showed that air pollution considerably affects children's respiratory morbidity, deserving attention from the health authorities. PMID- 15785836 TI - Autoimmunity and molecular mimicry in tropical spastic paraparesis/human T lymphotropic virus-associated myelopathy. AB - Viruses share antigenic sites with normal host cell components, a phenomenon known as molecular mimicry. It has long been suggested that viral infections might trigger an autoimmune response by several mechanisms including molecular mimicry. More than 600 antiviral monoclonal antibodies generated against 11 different viruses have been reported to react with 3.5% of cells specific for uninfected mouse organs. The main pathological feature of tropical spastic paraparesis/human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (TSP/HAM) is a chronic inflammation of the spinal cord characterized by perivascular cuffing of mononuclear cells accompanied by parenchymal lymphocytic infiltration. We detected the presence of autoantibodies against a 98- to 100-kDa protein of in vitro cultured human astrocytes and a 33- to 35-kDa protein from normal human brain in the serum of HTLV-I-seropositive individuals. The two cell proteins exhibited molecular mimicry with HTLV-I gag and tax proteins in TSP/HAM patients, respectively. Furthermore, the location of 33- to 35-kDa protein cross reaction correlated with the anatomical spinal cord areas (in the rat model) in which axonal damage has been reported in several cases of TSP/HAM patients. Our experimental evidence strongly suggests that the demyelinating process occurring in TSP/HAM may be mediated by molecular mimicry between domains of some viral proteins and normal cellular targets of the spinal cord sections involved in the neurodegeneration. PMID- 15785835 TI - Seroprevalence of human herpesvirus 8 infection in children born to HIV-1 infected women in Sao Paulo, Brazil. AB - Human herpesvirus-8 (HHV-8) appears to be transmitted mainly by sexual contact. However, several studies suggest that in developing countries the infection may be acquired early in life by routes other than sexual transmission. The present study estimated the seroprevalence of HHV-8 in Brazilian children born to HIV-1 infected mothers. The serum samples were collected in a cross-sectional cohort study from 99 children born to HIV-infected mothers (median age 3.27 years; range 1.5-13.8 years) attending the outpatient clinic of the Federal University of Sao Paulo. IgG antibodies to HHV-8 latency-associated nuclear antigen and lytic phase antigens were detected by immunofluorescence assays. The samples tested were collected from children aged 12 months or older to exclude the possibility of cross-placental antibody transport. The total prevalence of anti-lytic antibodies in this population (5/99; 5%) reveals that HHV-8 infection can occur during childhood. Children aged 1.5 to 2 years had a seroprevalence of 2% (1/50) and children aged 3.25 to 13.8 years had a seroprevalence of 8% (4/49). This difference was not statistically significant, probably because of the small size of the sample, but it suggests that HHV-8 infection occurs more commonly late in infancy. Further prospective studies are necessary to evaluate the timing and risk factors for primary HHV-8 infection in the pediatric population. PMID- 15785838 TI - Cultural adaptation and validation of the "Kidney Disease and Quality of Life- Short Form (KDQOL-SF 1.3)" in Brazil. AB - The objective of the present study was to translate the Kidney Disease Quality of Life -- Short Form (KDQOL-SF 1.3) questionnaire into Portuguese to adapt it culturally and validate it for the Brazilian population. The KDQOL-SF was translated into Portuguese and back-translated twice into English. Patient difficulties in understanding the questionnaire were evaluated by a panel of experts and solved. Measurement properties such as reliability and validity were determined by applying the questionnaire to 94 end-stage renal disease patients on chronic dialysis. The Nottingham Health Profile Questionnaire, the Karnofsky Performance Scale and the Kidney Disease Questionnaire were administered to test validity. Some activities included in the original instrument were considered to be incompatible with the activities usually performed by the Brazilian population and were replaced. The mean scores for the 19 components of the KDQOL-SF questionnaire in Portuguese ranged from 22 to 91. The components "Social support" and "Dialysis staff encouragement" had the highest scores (86.7 and 90.8, respectively). The test-retest reliability and the inter-observer reliability of the instrument were evaluated by the intraclass correlation coefficient. The coefficients for both reliability tests were statistically significant for all scales of the KDQOL-SF (P < 0.001), ranging from 0.492 to 0.936 for test-retest reliability and from 0.337 to 0.994 for inter-observer reliability. The Cronbach's alpha coefficient was higher than 0.80 for most of components. The Portuguese version of the KDQOL-SF questionnaire proved to be valid and reliable for the evaluation of quality of life of Brazilian patients with end-stage renal disease on chronic dialysis. PMID- 15785837 TI - Immunogenicity of a 23-valent pneumococcal polysaccharide vaccine in Brazilian elderly. AB - Serum antibodies specific for the capsular polysaccharides of Streptococcus pneumoniae provide protection against invasive pneumococcal infection. In Brazil, this vaccine has been used for people over 65 years with clinical risk to develop pneumococcal infection since 1999. We evaluated the immune response of 102 elderly subjects (75.5% females and 24.5% males) with a mean age of 71 years, and 19 young healthy adults (63.2% females and 36.8% males) with a mean age of 27 years. The elderly study group consisted of outpatients who received follow-up care in the Geriatric Department of General Hospital, Faculty of Medicine, University of Sao Paulo. None had acute illness at the time of vaccination. Both groups were immunized with one intra-deltoid injection with 0.5 ml of a 23-valent pneumococcal polysaccharide vaccine. The total IgG specific antibody concentrations to capsular polysaccharides 1, 3, 5, 6B, 8, and 14 were determined against pre- and 1-month post-vaccination sera. All samples were analyzed according to the second-generation pneumococcal polysaccharide ELISA protocol. We observed that the pneumococcal polysaccharide vaccine evoked consistent antibody increase for serotypes 1, 5, 6B, 8, and 14 (geometric mean concentration increase of 2.46 in the elderly and 2.84 in the young adults). Otherwise, we observed no increase in antibody concentration for serotype 3 in both groups. PMID- 15785840 TI - A study of the abilities in oral language comprehension of the Boston Diagnostic Aphasia Examination -- Portuguese version: a reference guide for the Brazilian population. AB - We analyzed the performance of 162 normal subjects, subdivided into groups according to age and schooling, in the oral comprehension tasks of the Boston Diagnostic Aphasia Examination translated and adapted to Brazilian Portuguese to obtain a profile of performance for the Brazilian population, as well as cut-off scores for each task, and to determine the best combination of tasks that distinguish normal from aphasic subjects, as a guide for clinicians. The normal subjects were compared to 69 aphasics. Age alone influenced the performance in the designation of actions (subjects above 70 years showing the worst performance); schooling alone influenced the comprehension of forms, colors and numbers (subjects with less than four years of education showing a poorer performance). Both age and schooling influenced the performance in Body Part Identification (BPI) and Complex Ideational Material (CIM) with mean values of 70.5 +/- 3.3 (Word Discrimination, WD), 18.9 +/- 1.4 (BPI), 14.7 +/- 0.9 (Commands), and 10.3 +/- 1.7 (CIM) for the whole sample; the cut-off scores obtained were 65 (WD), 17.5 (BPI), 14 (Commands), and 9.5 (CIM) for the whole sample. Logistic regression showed that the combination of BPI + Commands + CIM was the most efficient in differentiating normal subjects from aphasics, with 72.5% sensitivity and 97.6% specificity. However, for low-education subjects, BPI and Commands were sufficient for this differentiation (75.7% sensitivity and 84.7% specificity). The main contribution of this study was to provide reference values that are far more representative of our population to be used by health professionals in Brazil, taking into account cultural differences. PMID- 15785839 TI - Laminin concentration in ascites of patients with hepatic cirrhosis and peritoneal carcinomatosis. AB - Laminin levels in ascitic fluid have been proposed as a marker for neoplastic ascites. We compared the concentration of laminin in serum and in ascitic fluid from patients with hepatic cirrhosis and peritoneal carcinomatosis and assessed the diagnostic value of serum laminin levels in differentiating neoplastic from benign ascites. Laminin concentrations were determined by ELISA with antibodies against laminin extracted from the human placenta, in patients with ascites due to peritoneal carcinomatosis (N = 20) and hepatic cirrhosis (N = 33). Patients with infected or hemorrhagic ascites were excluded. The receiver operating characteristic curve was used to determine the sensitivity and specificity of serum laminin for the diagnosis of neoplastic ascites. When compared to the group with cirrhosis, the carcinomatosis group presented significantly higher mean laminin levels in serum (3.3 +/- 0.5 vs 2.1 +/- 0.4 microg/ml, mean +/- SD, P < 0.05) and ascites (2.8 +/- 0.5 vs 1.6 +/- 0.4 microg/ml, P < 0.05). Although laminin concentration was higher in serum than in ascites, the laminin serum/ascites ratio and serum-ascites gradient did not differ between the studied groups. A significant correlation (r = 0.93, P < 0.0001) was observed between the serum and ascites laminin values. Serum laminin levels >2.25 microg/ml showed 100% sensitivity and 73% specificity for the diagnosis of neoplastic ascites. Serum concentration seems to be the main determinant of laminin levels in ascitic fluid and its values can be used as a diagnostic parameter in the study of neoplastic ascites. PMID- 15785841 TI - Translation into Brazilian Portuguese, cultural adaptation and evaluation of the reliability of the Disabilities of the Arm, Shoulder and Hand Questionnaire. AB - The objective of the present study was to translate, adapt and validate a Brazilian Portuguese version of the Disabilities of the Arm, Shoulder and Hand (DASH) Questionnaire. The study was carried out in two steps. The first was to translate the DASH into Portuguese and to perform cultural adaptation and the second involved the determination of the reliability and validity of the DASH for the Brazilian population. For this purpose, 65 rheumatoid arthritis patients of either sex (according to the classification criteria of the American College of Rheumatology), ranging in age from 18 to 60 years and presenting no other diseases involving the upper limbs, were interviewed. The patients were selected consecutively at the rheumatology outpatient clinic of UNIFESP. The following results were obtained: in the first step (translation and cultural adaptation), all patients answered the questions. In the second step, Spearman's correlation coefficients for interobserver evaluation ranged from 0.762 to 0.995, values considered to be highly reliable. In addition, intraclass correlation coefficients ranged from 0.97 to 0.99, also highly reliable values. Spearman's correlation coefficients and the intraclass correlation coefficients obtained during intra-observer evaluation ranged from 0.731 to 0.937 and from 0.90 to 0.96, respectively, being highly reliable values. The Ritchie Index showed a weak correlation with Brazilian DASH scores, while the visual analog scale of pain showed a good correlation with DASH score. We conclude that the Portuguese version of the DASH is a reliable instrument. PMID- 15785843 TI - Bone mineral density of Brazilian girls with juvenile dermatomyositis. AB - We measured bone mineral density (BMD) in girls with juvenile dermatomyositis (JDM) considering multiple factors in order to determine if it could be used as a predictor of reduction in bone mass. A cross-sectional study of lumbar spine BMD (L2-L4) was conducted on 10 girls aged 7-16 years with JDM. A group of 20 age matched healthy girls was used as control. Lumbar spine BMD was measured by dual energy X-ray absorptiometry. Weight, height and pubertal Tanner stage were determined in all patients and controls. Duration of disease and mean daily and cumulative steroid doses were calculated for all patients on the basis of their medical charts. JDM activity was determined on the basis of the presence of muscle weakness, cutaneous vasculitis and/or elevation of serum concentration of one or more skeletal muscle enzymes. Seven patients demonstrated osteopenia or osteoporosis. Lumbar BMD was significantly lower in the JDM patients than the age matched healthy control girls (0.712 vs 0.878, respectively; Student t-test, P = 0.041). No significant correlation between BMD and age, height, Tanner stage, disease duration, corticosteroid use, or disease activity was observed in JDM girls, but a correlation was observed between BMD and weight (Pearson's correlation coefficient, r = 0.802). Patients with JDM may be at risk for a significant reduction in BMD that might contribute to further skeletal fragility. Our results suggest that reduced bone mass in JDM may be related to other intrinsic mechanisms in addition to steroid treatment and some aspects of the disease itself may contribute to this condition. PMID- 15785842 TI - Effects of chronic heart disease on skeletal muscle fiber size. AB - Size changes in muscle fibers of subjects with chronic heart disease (CHD) have been reported, although a consensus has not been achieved. The aims of the present study were to investigate a possible association between CHD and fiber size changes in the brachial biceps compared to subjects without heart disease. Forty-six muscle samples were obtained in autopsies of individuals (13 to 84 years) without neuromuscular disorders, 19 (10 males and 9 females) with, and 27 (14 males and 13 females) without CHD. In all cases muscle sections were stained with hematoxylin and eosin and processed for the visualization of myofibrillar ATPase activity. The lesser diameter of type 1 and type 2 fibers was obtained tracing their outlines (at least 150 fibers of each type per sample) onto an image analyzer connected to a computer. The results were analyzed statistically comparing males and females with and without CHD. Type 1 fiber mean lesser diameters were 51.51 and 54.52 microm in males (normal range 34-71 microm) and 45.65 and 55.42 microm in females (normal range 34-65 microm) without and with CHD, respectively; type 2 fibers measured 54.31, 58.23, 41.15, and 49.57 microm, respectively (normal range 36-79 microm for males and 32-59 microm for females). No significant difference in fiber size was detected in 24 males with and without CHD, while in 22 females there was a significant increase in size in those with cardiomyopathy. We concluded that CHD does not determine significant changes in fiber size. However, in females, there is some hypertrophy which, despite within normal range, may reflect morphologic heterogeneity of the sample, or the daily life activities in the upper limbs as a compensatory mechanism to fatigability that affect predominantly the lower limbs in subjects with CHD. PMID- 15785844 TI - Single nucleotide +1 frameshifts in an apparently functional mitochondrial cytochrome b gene in ants of the genus Polyrhachis. AB - Twelve of 30 species examined in the ant genus Polyrhachis carry single nucleotide insertions at one or two positions within the mitochondrial cytochrome b (cytb) gene. Two of the sites are present in more than one species. Nucleotide substitutions in taxa carrying insertions show the strong codon position bias expected of functional protein coding genes, with substitutions concentrated in the third positions of the original reading frame. This pattern of evolution of the sequences strongly suggests that they are functional cytb sequences. This result is not the first report of +1 frameshift insertions in animal mitochondrial genes. A similar site was discovered in vertebrates, where single nucleotide frameshift insertions in many birds and a turtle were reported by Mindell et al. (Mol Biol Evol 15:1568, 1998). They hypothesized that the genes are correctly decoded by a programmed frameshift during translation. The discovery of four additional sites gives us the opportunity to look for common features that may explain how programmed frameshifts can arise. The common feature appears to be the presence of two consecutive rare codons at the insertion site. We hypothesize that the second of these codons is not efficiently translated, causing a pause in the translation process. During the stall the weak wobble pairing of the tRNA bound in the peptidyl site of the ribosome, together with an exact Watson-Crick codon-anticodon pairing in the +1 position, allows translation to continue in the +1 reading frame. The result of these events is an adequate level of translation of a full-length and fully functional protein. A model is presented for decoding of these mitochondrial genes, consistent with known features of programmed translational frameshifting in the yeast TY1 and TY3 retrotransposons. PMID- 15785845 TI - Phylogenetic analyses of the core antenna domain: investigating the origin of photosystem I. AB - Phototrophy, the conversion of light to biochemical energy, occurs throughout the Bacteria and plants, however, debate continues over how different phototrophic mechanisms and the bacteria that contain them are related. There are two types of phototrophic mechanisms in the Bacteria: reaction center type 1 (RC1) has core and core antenna domains that are parts of a single polypeptide, whereas reaction center type 2 (RC2) is composed of short core proteins without antenna domains. In cyanobacteria, RC2 is associated with separate core antenna proteins that are homologous to the core antenna domains of RC1. We reconstructed evolutionary relationships among phototrophic mechanisms based on a phylogeny of core antenna domains/proteins. Core antenna domains of 46 polypeptides were aligned, including the RC1 core proteins of heliobacteria, green sulfur bacteria, and photosystem I (PSI) of cyanobacteria and plastids, plus core antenna proteins of photosystem II (PSII) from cyanobacteria and plastids. Maximum likelihood, parsimony, and neighbor joining methods all supported a single phylogeny in which PSII core antenna proteins (PsbC, PsbB) arose within the cyanobacteria from duplications of the RC1-associated core antenna domains and accessory antenna proteins (IsiA, PcbA, PcbC) arose from duplications of PsbB. The data indicate an evolutionary history of RC1 in which an initially homodimeric reaction center was vertically transmitted to green sulfur bacteria, heliobacteria, and an ancestor of cyanobacteria. A heterodimeric RC1 (=PSI) then arose within the cyanobacterial lineage. In this scenario, the current diversity of core antenna domains/proteins is explained without a need to invoke horizontal transfer. PMID- 15785847 TI - Multiple independent losses of photosynthesis and differing evolutionary rates in the genus Cryptomonas (Cryptophyceae): combined phylogenetic analyses of DNA sequences of the nuclear and the nucleomorph ribosomal operons. AB - In this study, evidence for at least three independent losses of photosynthesis in the freshwater cryptophyte genus Cryptomonas is presented. The phylogeny of the genus was inferred by molecular phylogenetic analyses of the nuclear internal transcribed spacer 2 (nuclear ITS2), partial nuclear large subunit ribosomal DNA (LSU rDNA), and nucleomorph small subunit ribosomal DNA (SSU rDNA, NM). Both concatenated and single data sets were used. In all data sets, the colorless Cryptomonas strains formed three different lineages, always supported by high bootstrap values (maximum parsimony, neighbor joining and maximum likelihood) and posterior probabilities (Bayesian analyses). The three leukoplast-bearing lineages displayed differing degrees of accelerated evolutionary rates in nuclear and nucleomorph rDNA. Also an increase in A + T-content in highly variable regions of the nucleomorph SSU rDNA was observed in one of the leukoplast-bearing lineages. PMID- 15785846 TI - Sequence, transcription activity, and evolutionary origin of the R-body coding plasmid pKAP298 from the intracellular parasitic bacterium Caedibacter taeniospiralis. AB - We isolated the intracellular parasitic bacterium Caedibacter taeniospiralis from cultures of the freshwater ciliate Paramecium tetraurelia strain 298. Plasmid pKAP298 as well as the total RNA were isolated from the bacteria. pKAP298 was totally sequenced (49.1 kb; NCBI accession number AY422720). From southern blots of pKAP-fragments and Digoxigenin-labeled cDNA of the Caedibacter-RNA, we generated transcription maps of pKAP298. The observed transcription activity indicated functions of the plasmid besides the synthesis of the R-body, a complex protein inclusion associated with toxic effects of Caedibacter cells on host paramecia. We identified 63 potential protein coding regions on pKAP298, and a novel transposon as well as known transposons were characterized. A group II intron was identified. Homologies with putative phage genes were detected on pKAP298 that direct to the evolution of pKAP298 from a bacteriophage. This original phage most probably belonged to the Caudovirales. Hints on a toxin coding region of pKAP298 are given: a protein with homology to the Soj-/ParA family also has homologies to a membrane associated ATPase, which is involved in eukaryotic ATPase dependent ion carriers and may be associated with toxic effects on paramecia ingesting this protein. PMID- 15785848 TI - A novel additional group II intron distinguishes the mitochondrial rps3 gene in gymnosperms. AB - Comparative analysis of the ribosomal protein S3 gene (rps3) in the mitochondrial genome of Cycas with newly sequenced counterparts from Magnolia and Helianthus and available sequences from higher plants revealed that the positional clustering with the genes for ribosomal protein S19 (rps19) and L16 (rpl16) is preserved in gymnosperms. However, in contrast to the other land plant species, the rps3 gene in Cycas mitochondria is unique in possessing a second intron: rps3i2. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of the transcripts generated from the rps19-rps3-rpl16 cluster in Cycas mitochondria demonstrated that the genes are cotranscribed and extensively modified by RNA editing and that both introns are efficiently spliced. Despite remarkable size heterogeneity, the Cycas rps3i1 can be shown to be homologous to the group IIA introns present within the rps3 gene of algae and land plants, including Magnolia and Helianthus. Conversely, sequences similar to the rps3i2 have not been reported previously. On the basis of conserved primary and secondary structure the second intervening sequence interrupting the Cycas rps3 gene has been classified as a group II intron. The close relationship of the rps3i2 to a group of different plant mitochondrial introns is intriguing and suggestive of a mitochondrial derivation for this novel intervening sequence. Interestingly, the rps3i2 appears to be conserved at the same gene location in other gymnosperms. Furthermore, the pattern of the rps3i2 distribution among algae and land plants provides evidence for the evolutionary acquisition of this novel intron in gymnosperms via intragenomic transposition or retrotransposition. PMID- 15785849 TI - Multiple mutations and gene duplications conferring organophosphorus insecticide resistance have been selected at the Rop-1 locus of the sheep blowfly, Lucilia cuprina. AB - Sequences of the esterase gene alpha E7 were compared across 41 isogenic (IV) strains of the sheep blowfly, Lucilia cuprina, and one strain of the sibling species, L. sericata. The 1.2-kb region sequenced includes sites of two insecticide resistance mutations. Gly137Asp confers resistance to organophosphorus insecticides (OPs), particularly preferring diethyl OPs such as diazinon, while Trp251Leu prefers dimethyl OPs, and particularly malathion, with the additional presence of carboxylester moieties. We found that there are just eight haplotypes among the 41 chromosomes studied: two Gly137Asp containing haplotypes, two Trp251Leu containing haplotypes, and four susceptible haplotypes, including the L. sericata sequence. While phylogenetic analysis of these haplotypes suggests that the Asp137 and Leu251 mutations each arose at least twice, evidence for recombination was detected across the region, therefore single origins for these resistance mutations cannot be ruled out. Levels of linkage disequilibrium in the data are high and significant hitchhiking is indicated by Fay and Wu' s H test but not the Tajima test. A test of haplotype diversity indicates a paucity of diversity compared with neutral expectations. Both these results are consistent with a very recent selective sweep at the Lc alphaE7 locus. Interestingly, gene duplications of three different combinations of OP resistant haplotypes were identified in seven of the isogenic (IV) strains. All three types of duplication involve an Asp137 and a Trp251 haplotype. To examine whether more haplotypes existed before the hypothesised selective sweep, fragments of alpha E7 surrounding the resistance mutations were amplified from pinned material dating back to before OPs were used. Four new sequence haplotypes, not sampled in the survey of extant haplotypes, were obtained that are all associated with susceptibility. This is suggestive of a higher historical level of susceptible allelic diversity at this locus. PMID- 15785850 TI - Natural selection during functional divergence to LMP7 and proteasome subunit X (PSMB5) following gene duplication. AB - The LMP7 and PSMB5 genes were created through an ancient gene duplication event of their ancestral locus. These proteins contain an active site of proteolysis, and LMP7 replaces PSMB5 as a component of the 20S proteasome after stimulation of cells by interferon-gamma. Replacement of PSMB5 by LMP7 changes the profile of the products of 20S proteasome processing, predisposing digested peptides for transport to and display by the immune system. The purpose of this study is to investigate evolutionary forces influencing functional divergence between LMP7 and PSMB5 following duplication. Levels of synonymous and nonsynonymous substitution rates are estimated to infer differences in levels of natural selection. Estimates of substitution rates indicate that natural selection elevated rates of nonsynonymous substitution in LMP7 following gene duplication, whereas PSMB5 experienced an increase in substitution rate that was not likely due to diversifying natural selection following duplication. Following initial divergence, nearly neutral mutations have dominated gene evolution in both lineages. The LMP7 gene locus provides a rare example of a protein with specialized function arising from duplication and divergence of a housekeeping protein by way of natural selection. PMID- 15785851 TI - Evolution of NIN-like proteins in Arabidopsis, rice, and Lotus japonicus. AB - Genetic studies in Lotus japonicus and pea have identified Nin as a core symbiotic gene required for establishing symbiosis between legumes and nitrogen fixing bacteria collectively called Rhizobium. Sequencing of additional Lotus cDNAs combined with analysis of genome sequences from Arabidopsis and rice reveals that Nin homologues in all three species constitute small gene families. In total, the Arabidopsis and rice genomes encode nine and three NIN-like proteins (NLPs), respectively. We present here a bioinformatics analysis and prediction of NLP evolution. On a genome scale we show that in Arabidopsis, this family has evolved through segmental duplication rather than through tandem amplification. Alignment of all predicted NLP protein sequences shows a composition with six conserved modules. In addition, Lotus and pea NLPs contain segments that might characterize NIN proteins of legumes and be of importance for their function in symbiosis. The most conserved region in NLPs, the RWP-RK domain, has secondary structure predictions consistent with DNA binding properties. This motif is shared by several other small proteins in both Arabidopsis and rice. In rice, the RWP-RK domain sequences have diversified significantly more than in Arabidopsis. Database searches reveal that, apart from its presence in Arabidopsis and rice, the motif is also found in the algae Chlamydomonas and in the slime mold Dictyostelium discoideum. Thus, the origin of this putative DNA binding region seems to predate the fungus-plant divide. PMID- 15785852 TI - Phylogenetic and exon-intron structure analysis of fungal subtilisins: support for a mixed model of intron evolution. AB - Phylogenetic and exon-intron structure analyses of intra- and interspecific fungal subtilisins in this study provided support for a mixed model of intron evolution: a synthetic theory of introns-early and introns-late speculations. Intraspecifically, there were three phase zero introns in Pr1A and its introns 1 and 2 located at the highly conserved positions were phylogentically congruent with coding region, which is in favor of the view of introns-early speculation, while intron 3 had two different sizes and was evolutionarily incongruent with coding region, the evidence for introns-late speculation. Noticeably, the subtilisin Pr1J gene from different strains of M. ansiopliae contained different number of introns, the strong evidence in support of introns-late theory. Interspecifically, phylogenetic analysis of 60 retrievable fungal subtilisins provided a clear relationship between amino acid sequence and gene exon-intron structure that the homogeneous sequences usually have a similar exon-infron structure. There were 10 intron positions inserted by highly biased phase zero introns across examined fungal subtilisin genes, half of these positions were highly conserved, while the others were species-specific, appearing to be of recent origins due to intron insertion, in favor of the introns-late theory. High conservations of positions 1 and 2 inserted by the high percentage of phase zero introns as well as the evidence of phylogenetic congruence between the evolutionary histories of intron sequences and coding region suggested that the introns at these two positions were primordial. PMID- 15785853 TI - Expression patterns of duplicate genes in the developing root in Arabidopsis thaliana. AB - Data on gene expression in the development of the root in Arabidopsis thaliana were used to test for expression profile differences among multi-gene families and to examine the extent to which expression differences accompanied coding sequences divergence within families. Significant differences among families were observed on two principal axes, accounting for over 80% of the variance in the expression data. The number of synonymous nucleotide substitutions per synonymous site (d(S)) and the number of nonsynonymous nucleotide substitutions per nonsynonymous site (d(N)) were estimated between the members of two-member families (N = 428) and between phylogenetically independent sister pairs (N = 190) of sequences within larger families. Ribosomal proteins and a few other proteins were exceptional in showing highly divergent expression patterns in spite of very low levels of amino acid sequence divergence, as indicated by the low d(N) relative to d(S). However, the majority of gene duplicates showed relatively high levels of amino acid sequence divergence without appreciable change in expression pattern in the cell types analyzed. PMID- 15785855 TI - Evidence of positive Darwinian selection in Omp85, a highly conserved bacterial outer membrane protein essential for cell viability. AB - Omp85 is a highly conserved outer membrane protein found in all gram-negative bacteria. It is essential for bacterial cell viability and plays an integral function in the positioning and folding of other outer membrane proteins into the bacterial outer membrane. We have employed a maximum likelihood and a maximum parsimony approach to detect evidence of positive Darwinian selection in Omp85 homologues from 10 delta-proteobacteria and have identified 14 amino acid sites that show evidence of being under the influence of adaptive evolution. Interestingly all sites bar one are concentrated within surface loops of the protein that most likely interact with host immune response or the surrounding environment. Alternatively amino acids within membrane-spanning regions of the protein are found to be under purifying selection most likely as a result of structural constraints. PMID- 15785854 TI - Evolution of plant-like crystalline storage polysaccharide in the protozoan parasite Toxoplasma gondii argues for a red alga ancestry. AB - Single-celled apicomplexan parasites are known to cause major diseases in humans and animals including malaria, toxoplasmosis, and coccidiosis. The presence of apicoplasts with the remnant of a plastid-like DNA argues that these parasites evolved from photosynthetic ancestors possibly related to the dinoflagellates. Toxoplasma gondii displays amylopectin-like polymers within the cytoplasm of the dormant brain cysts. Here we report a detailed structural and comparative analysis of the Toxoplasma gondii, green alga Chlamydomonas reinhardtii, and dinoflagellate Crypthecodinium cohnii storage polysaccharides. We show Toxoplasma gondii amylopectin to be similar to the semicrystalline floridean starch accumulated by red algae. Unlike green plants or algae, the nuclear DNA sequences as well as biochemical and phylogenetic analysis argue that the Toxoplasma gondii amylopectin pathway has evolved from a totally different UDP-glucose-based metabolism similar to that of the floridean starch accumulating red alga Cyanidioschyzon merolae and, to a lesser extent, to those of glycogen storing animals or fungi. In both red algae and apicomplexan parasites, isoamylase and glucan-water dikinase sequences are proposed to explain the appearance of semicrystalline starch-like polymers. Our results have built a case for the separate evolution of semicrystalline storage polysaccharides upon acquisition of photosynthesis in eukaryotes. PMID- 15785856 TI - On slippage-like mutation dynamics within genes: a study of pseudogenes and 3'UTRs. PMID- 15785857 TI - Size frequency distribution of prion protein (PrP) aggregates in variant Creutzfeldt-Jakob disease (vCJD). AB - The frequency distribution of aggregate size of the diffuse and florid-type prion protein (PrP) plaques was studied in various brain regions in cases of variant Creutzfeldt-Jakob disease (vCJD). The size distributions were unimodal and positively skewed and resembled those of beta-amyloid (A beta) deposits in Alzheimer's disease (AD) and Down's syndrome (DS). The frequency distributions of the PrP aggregates were log-normal in shape, but there were deviations from the expected number of plaques in specific size classes. More diffuse plaques were observed in the modal size class and fewer in the larger size classes than expected and more florid plaques were present in the larger size classes compared with the log-normal model. It was concluded that the growth of the PrP aggregates in vCJD does not strictly follow a log-normal model, diffuse plaques growing to within a more restricted size range and florid plaques to larger sizes than predicted. PMID- 15785858 TI - Increased cell-cell adhesion, a novel effect of R-(-)-deprenyl. AB - The neuroprotective effect of the antiparkinsonian monoamine oxidase (MAO)-B inhibitor, R-(-)-deprenyl has been under investigation for years. Cytoskeleton, a main component of cell adhesion, is involved in the development of R-(-)-deprenyl responsive diseases, the effect of the drug on cell adhesion, however, is not known. We examined the effect of R-(-)-deprenyl on cell-cell adhesion of neuronal and non-neuronal cells. R-(-)-deprenyl treatment resulted in a cell type- and concentration-dependent increase in cell-cell adhesion of PC12 and NIH3T3 cells at concentrations lower than those required for MAO-B inhibition, while S-(+) deprenyl was not effective. This acitvity of R-(-)-deprenyl was not prevented by the cytochrome P-450 inhibitor, SKF525A, while deprenyl-N-oxide, a newly described metabolite, also induced an increase in cell-cell adhesion. The effect of R-(-)-deprenyl was not reversible during a 24-hour recovery period. In summary, we described a new, MAO-B independent effect of R-(-)-deprenyl on cell cell adhesion which can contribute to its neuroprotective function. PMID- 15785859 TI - Expression levels of Rab2, a G protein, and Bag-1, a Bcl-2 binding protein are controlled by withdrawal of nicotine from cultured pheochromocytoma PC12 cells. AB - We previously reported that nicotine withdrawal up-regulates transcription of some immediately early genes (IEGs), c-fos (Ichino et al., 1999) and egr1, nur77 (Ichino et al., 2002) in cultures of pheochromocytoma PC12 cells, which are of neuronal lineage. In the present study we aimed at further elucidating the effects of nicotine withdrawal on the expression of the genes downstream of IEGs. We examined the changes in the protein levels of 2 GTP-binding proteins, Rab2 (Ras-related protein) and Rac1. PC12 cells were cultured in the presence of nicotine for 24 hours, and then the nicotine was removed from the medium. The protein level of Rab2 was low in the presence of nicotine, but was rapidly increased after nicotine withdrawal. In contrast, that of Rac1 did not change after the withdrawal. Considering the neuroprotective effect of nicotine, we also examined the level of Bag-1 protein, which is a binding protein for Bcl-2, an anti-apoptotic factor, and found a slight increase in the gene expression of Bag 1 following nicotine withdrawal. Among 56-kDa, 50-kDa, and 36-kDa protein components of the Bag-1 protein complex, the levels of 56-kDa and 50-kDa proteins were not changed by the addition or withdrawal of nicotine; but the level of the 36-kDa protein, which had been increased in the presence of nicotine, was markedly decreased after nicotine withdrawal. The present results suggest that such changes may also occur in individuals during abstaining from smoking and be related to the withdrawal symptoms experienced after smokers stop smoking. PMID- 15785860 TI - An association study of dopamine receptors polymorphisms and the Wisconsin Card Sorting Test in schizophrenia. AB - Dopamine (DA), an important neurotransmitter in prefrontal cortex (PFC), is involved in the pathogenesis of schizophrenia. The aim of the study was to test an association between common polymorphism of genes for DA receptors DRD1, DRD2, DRD3, DRD4, and performance on the Wisconsin Card Sorting Test (WCST), measuring various functions of PFC, in 138 schizophrenic patients. Patients with G/G genotype of DRD1 tended to obtain worse results in all domains of WCST compared to patients with remaining genotypes, particularly for number of completed corrected categories, and trials to set the first category. A relationship was also found in female patients between DRD2 polymorphism and number of perseverative errors, while no association between WCST results and DRD3 or DRD4 polymorphism was observed in patients studied. The results may suggest an association between DRD1 gene polymorphism and performance on PFC test in schizophrenia. Also, the gender-dependent role of DRD2 in this process may be presumed. PMID- 15785861 TI - Clinical and genetic evaluation of 8 Polish families with levodopa-responsive parkinsonism. AB - We studied 8 large Polish families with parkinsonism, 6 of which were newly identified. Thirty-six family members had well-documented levodopa-responsive parkinsonism. The phenotype of affected individuals was indistinguishable from that of persons with idiopathic Parkinson disease (PD). The pattern of inheritance in 5 families was consistent with autosomal dominant transmission; in 3 families the mode of inheritance was uncertain. Single photon emission computed tomography (SPECT) studies with the dopamine transporter radioligand [(123)I]FP CIT were performed in 1 family. The SPECT study showed striatal presynaptic dopaminergic degeneration consistent with sporadic PD in 1 affected family member and no signs of nigrostriatal dopaminergic dysfunction in 5 at-risk individuals. Sequence analysis in all 8 families excluded known genes associated with familial parkinsonism. Genome-wide 2-point linkage studies in the largest 2 families did not identify significant linkage (z > 3.0), although positive scores were obtained for 5q23 (D5S1462 and D5S2501), a locus previously implicated in disease susceptibility. PMID- 15785862 TI - Retrospective evaluation of cardio-pulmonary fibrotic side effects in symptomatic patients from a group of 234 Parkinson's disease patients treated with cabergoline. AB - BACKGROUND: Cardiac valvulopathy has been recently associated with the use of the ergot dopamine agonist (EDA) pergolide in Parkinson's disease (PD). Cabergoline a widely used, well-tolerated EDA which has also been recently implicated in relation to fibrotic side effects although the evidence base for this is not sound. AIMS: In PD patients on chronic cabergoline therapy, do symptoms suggestive of serosal/cardiac fibrosis imply underlying fibrotic lesions? METHODS: A retrospective data review of 234 PD cases from three UK centres, on chronic cabergoline monotherapy or adjunctive treatment to identify symptoms suggestive of pleuro-pulmonary, cardiac or retroperitoneal fibrosis. These causes were thereafter selectively examined by appropriate specialists with relevant investigations. CONCLUSIONS: Out of 234 cases, 15 were identified with symptoms suggestive of respiratory, cardiac or abdominal systems involvement although subsequent investigations failed to reveal definite association with cabergoline except two cases with probable alveolitis and a possible association with cardiac murmur in one case. In spite of the deficiencies of a retrospective study, the results suggest a low risk of fibrotic side effects with cabergoline, particularly cardiac valvulopathy. PMID- 15785863 TI - Spinocerebellar ataxias types 2 and 3: degeneration of the pre-cerebellar nuclei isolates the three phylogenetically defined regions of the cerebellum. AB - The pre-cerebellar nuclei act as a gate for the entire neocortical, brainstem and spinal cord afferent input destined for the cerebellum. Since no pathoanatomical studies of these nuclei had yet been performed in spinocerebellar ataxia type 2 (SCA2) or type 3 (SCA3), we carried out a detailed postmortem study of the pre cerebellar nuclei in six SCA2 and seven SCA3 patients in order to further characterize the extent of brainstem degeneration in these ataxic disorders. By means of unconventionally thick serial sections through the brainstem stained for lipofuscin pigment and Nissl material, we could show that all of the pre cerebellar nuclei (red, pontine, arcuate, prepositus hypoglossal, superior vestibular, lateral vestibular, medial vestibular, interstitial vestibular, spinal vestibular, vermiform, lateral reticular, external cuneate, subventricular, paramedian reticular, intercalate, interfascicular hypoglossal, and conterminal nuclei, pontobulbar body, reticulotegmental nucleus of the pons, inferior olive, and nucleus of Roller) are among the targets of both of the degenerative processes underlying SCA2 and SCA3. These novel findings are in contrast to the current neuropathological literature, which assumes that only a subset of pre-cerebellar nuclei in SCA2 and SCA3 may undergo neurodegeneration. Widespread damage to the pre-cerebellar nuclei separates all three phylogenetically and functionally defined regions of the cerebellum, impairs their physiological functions and thus explains the occurrence of gait, stance, limb and truncal ataxia, dysarthria, truncal and postural instability with disequilibrium, impairments of the vestibulo-ocular reaction and optokinetic nystagmus, slowed and saccadic smooth pursuits, dysmetrical horizontal saccades, and gaze-evoked nystagmus during SCA2 and SCA3. PMID- 15785864 TI - Quality of life and gender identity in Parkinson's disease. AB - We examined the correlation between gender identity (perception of masculinity or femininity) and quality of life (QoL) of 124 Parkinson's disease (PD) patients without dementia (69 men, 55 women, mean age 65.8+/-10.2 years, mean disease duration 8.5+/-5.8 years, mean Hoehn and Yahr [H&Y] stage 2.7+/-0.8). All patients underwent clinical examinations and completed the PDQ-39 and the quality of sexual life questionnaire. Their masculine or feminine stereotypes were determined by the Bem Sex Role Inventory (BSRI) modified by Dior. QoL was significantly correlated with disease duration (r=0.262, p<0.01), H&Y staging (r=0.330, p<0.001) and disease severity (UPDRS) (r=0.432, p<0.001). The QoL of androgynous men and women (i.e., with strong feminine and masculine characteristics) was significantly (p<0.05) better than the other gender groups. A significant interaction was found between the sexes to gender identity (p<0.05). CONCLUSION: Androgynous PD patients cope better with their disease in terms of QoL parameters, especially androgynous women. PMID- 15785865 TI - Intrathecal administration of doxepin attenuated development of formalin-induced pain in rats. AB - The aim of the present research was to assess the influence of a tricyclic antidepressant doxepin administered intrathecally (i.t.) on the pain behavior in the formalin test (100 microl of 12% formalin was injected into the dorsal part of the hind paw under halotane anesthesia) in male Wistar rats. The influence of doxepin (62.5 microg i.t.) on the pain threshold and number of formalin-induced pain behaviors, as well as antinociceptive effect of morphine was studied. Doxepin significantly increased the nociceptive threshold in the paw pressure test, reduced formalin-induced pain behavior and potentiated morphine antinociceptive effect in formalin test. The obtained results indicate that analgesic effect of doxepin used before the injury is observable at the spinal level after intrathecal treatment, but not only after peripheral administration, which was shown in our previous study. The results of the present research demonstrated a possibility to modify the spinal nociceptive process by administration of doxepin before the formalin injection. PMID- 15785868 TI - Preface. PMID- 15785866 TI - Dopaminergic function in a family with the PARK6 form of autosomal recessive Parkinson's syndrome. AB - A G309D mutation in the PINK1 gene in a consanguineous Spanish kindred with seven siblings, three of whom are clinically affected, has recently been shown to be a cause of the PARK6 form of autosomal-recessive Parkinson's syndrome. In this family, we studied pre- and postsynaptic dopaminergic function using 123I-FP-CIT- and 123I-iodobenzamide-SPECT to determine binding to the presynaptic dopamine transporter (DAT) and postsynaptic D2 receptors respectively. All three PARK6 patients showed reduced striatal DAT binding with posterior preponderance similar to sporadic idiopathic PD, but only one patient showed significant striatal asymmetry. In two of the siblings, DAT binding was markedly increased. IBZM-SPECT was normal in both patients and sibs. Our findings indicate that 123I-FP-CIT SPECT shows similar DAT binding in PARK6 patients compared to idiopathic Parkinson's disease. The increased DAT binding in heterozygous PARK6 carriers may be a new very early preclinical finding, but its significance is still unclear. PMID- 15785869 TI - Epidemiology of head and neck cancer: magnitude of the problem. AB - Head and neck cancer comprises squamous cell carcinomas of the upper aerodigestive tract. There are similarities in their natural history, epidemiology and control. For these cancers premalignant changes can be identified. Smoking and drinking are the major risk factors. The geographical variations in incidence and mortality are indicative of differences in the prevalence of risk factors between countries. The dramatic increase in head and neck cancers is cause for great concern, particularly in Central-Eastern Europe. The great majority of these cancers could be prevented by reducing the prevalence of established risk factors. Screening could be used to detect both precancerous lesions and early invasive cancers; however, no study as yet has demonstrated a reduced incidence and mortality resulting from screening. When setting strategies for prevention, the socioeconomic differentials in incidence and mortality from head and neck cancers need to be taken into account. PMID- 15785870 TI - Human papillomavirus in head and neck cancer: molecular biology and clinicopathological correlations. AB - Human papillomaviruses are known to cause cancers of the cervix and other anogenital tract sites. Epidemiologic and molecular pathology studies have also suggested that HPV infection may be associated with cancers of the head and neck. Modes of transmission of HPV infection in the head and neck region have not been fully resolved; however, perinatal transmission and an association between sexual behavior and risk for HPV-positive cancers have been presented. Among the HPV types infecting the mucosa, high-risk, intermediate-risk and low-risk genotypes are defined, depending on their presence in carcinoma or precursor lesions. The phylogenic groups of HPVs also showed a definite correlation with the morphology of head and neck tumors. The groups A6, A7, and A9 include viruses that are frequently demonstrated in basaloid and verrucosus squamous cell carcinomas known to associate with HPV infection. Integration of HPV DNA into the host cell genome occurs early in cancer development and is an important event in malignant transformation. There is a trend for patients with HPV-positive tumors to be nondrinkers or light drinkers, the majority of these patients are females, and the median age is lower than in the case of HPV-negative tumors, but this latter difference was not always statistically significant. In the Kaplan-Meier survival model, the HPV-positive verrucous and basaloid squamous cell carcinomas showed better survival rates than the HPV-negative typical squamous cell carcinomas. An increased radiocurability of HPV-positive head and neck squamous cell carcinoma (HNSCC) has also been demonstrated. PMID- 15785871 TI - Tumor cell invasion and survival in head and neck cancer. AB - Squamous cell carcinoma (SCC) is the primary tumor type in head and neck cancer. Typically, these tumor cells show persistent invasion that frequently leads to local recurrence and distant lymphatic metastasis. The process of invasion involves concurrent infiltration and destruction of adjacent tissues. As with normal mucosal epithelium, SCC cells express receptors that mediate cell extracellular matrix (ECM) adhesion (integrins) and cell-cell adhesion (cadherins). Both receptor families represent important signaling devices that are capable of promoting survival and proliferation. Recent results indicate that integrins and cadherins cooperate to regulate invasive behavior. During SCC invasion, cells actively migrate through the surrounding ECM with the simultaneous remodeling of their intercellular adhesions. During invasion, integrin receptor engagement with specific ECM ligands along with concurrent remodeling of cadherin adhesions induces changes in the cytoskeleton though modulation of the activities of Rho family members. Tumor development and progression of SCC proceeds with the generation of variant cells with potential alterations in expression of adhesion receptors, and their associated signaling pathways lead to a highly invasive and metastatic phenotype. Understanding the molecular events that define this subset of invasive cells will facilitate the development of new treatment strategies. PMID- 15785872 TI - Biological significance of c-erbB family oncogenes in head and neck cancer. AB - Squamous cell carcinoma of the head and neck (SCCHN) tends to run an aggressive course and the prognosis has remained virtually unchanged in recent decades. The development of novel therapeutic strategies to improve patient outcome centres on the biology of the disease, namely the pivotal c-erbB family of growth factor receptors. c-erbB1 (or epidermal growth factor receptor, EGFR), is key to the pathogenesis of SCCHN and plays a central role in a complex network of downstream integrated signalling pathways. EGFR overexpression, detected in up to 90% of SCCHN, correlates with an increased risk of locoregional tumour relapse following primary therapy and relative resistance to treatment. The biological sequelae of erbB receptor activation are not simply cell proliferation, but also inhibition of apoptosis, enhanced migration, invasion, angiogenesis and metastasis: the 'hallmarks of cancer' [1]. As EGFR overexpression is associated with a poor clinical outcome in SCCHN, this receptor is attractive as a therapeutic target and the successful development of targeted therapies represents a paradigm shift in the medical approach to head and neck cancer. However, the extensive cross talk between signalling pathways, the multiple molecular aberrations and genetic plasticity in SCCHN all contribute to inherent and acquired resistance to both conventional and novel therapies. Understanding the cancer cell biology, in particular the significance of co-expression of c-erbB (and other) receptors, and the cell survival stimuli from (for example) activation of the phosphoinositide 3 kinase (PI3-kinase) cascade is fundamental to overcome current limitations in biologically targeted therapies. PMID- 15785873 TI - Molecular markers for diagnosis and prognosis. AB - A plethora of aberrations are associated with progress and outcome for head and neck cancer patients and some have been shown to provide prognostic information independent of the TNM staging system. These findings justify future studies that will harness recent advances in technologies to refine the range of molecular markers available. Important lessons have been learnt during the last two decades, particularly the need to perform multivariate analysis and to combine information derived from several molecular markers. At present the overall evidence is insufficient to alter clinical practice or to consider restricting clinical trials of new adjuvant to subsets of patients, identified on the basis of the use of molecular markers. However, a number of key prospective clinical studies are in progress and we can be optimistic that in the near future clinical research incorporating specific markers may provide new criteria for defining risk of relapse or of developing a new primary tumour, and guide the choice of optimal adjuvant treatment after surgery. The only way to truly validate a marker is to incorporate it into large clinical trials, and it is anticipated that the new treatment options that are on the horizon, that are likely to be an improvement on radiotherapy alone, will provide opportunities to help translate this laboratory research into clinical practice. PMID- 15785874 TI - Gene profiling in squamous cell carcinoma of the head and neck. AB - Survival for patients with squamous cell carcinoma of the head and neck (SCCHN) is still poor, despite great technical improvements in radiotherapy and surgery. A possible explanation for this is the lack of individualization in treatment based on biological properties of the tumors, resulting in over- as well as under treatment. Management of SCCHN has mainly been based on TNM classification over the last decades. However, a large amount of studies have shown that biomarkers may add prognostic information, independently of the TNM system, indicating that biological aggressiveness is not entirely reflected by the T- and N-status of the tumor. A conclusion to draw from this is that the present standardized treatment based on macroscopic features of the tumor in many cases will result in suboptimal treatment since important underlaying genetic properties of the tumors are not taken into consideration. A variety of laboratory techniques have been used in studies that investigate the individual biological features, spanning from methods that screen the genome for chromosomal and genetic abnormalities, e.g. cytogenetics, CGH, SKY, cDNA micro array to detailed studies of specific aberrations, e.g. southern, northern and western blotting, PCR based analysis and immunohistochemistry. Dysregulation of genes involved in e.g. cell cycle control, proliferation, drug resistance, and metastasis have been linked to outcome of treatment and survival. The purpose of this review of the literature was to summarize what has been studied so far by cDNA micro array techniques with regards to genetic screening in general and biomarkers that relate to response to therapy and prediction of clinical outcome in particular. We conclude that the majority of investigations that focus on gene profiling have a descriptive character, e.g. comparisons of tumor and normal cells, metastatic and non metastatic properties, and differences between sub-sites and grades of differentiation. There are just a handful studies that so far have investigated how gene profiling can be used to predict clinical course. PMID- 15785876 TI - Progression of head and neck squamous cell cancer. AB - Squamous cell cancer in the head and neck region (HNSC) is unique concerning its progression since it remains locoregional for long time and visceral metastases develop only in a later stage of the disease. Accordingly, molecular markers of the local invasion and the lymphatic dissemination both have critical importance. HNSC progression is associated with deregulated control of cell proliferation and apoptosis but it seems equally significant the disregulation of the proteolytic machineries. Here we outline the lymphatic metastatic cascade for HNSC to depict key molecular determinants as possible prognostic factors or therapeutic targets identifying immunological selection as a major feature. Unlike in local spreading, invasive potential of cancer cells seems to be less significant during lymphatic dissemination due to the anatomical properties of the lymphatic vessels and tissues. There is a general believe that HNSC is one disease however, data indicate that the anatomical localization of the tumor (the "soil") such as oral, lingual, glottic or pharyngeal has a significant effect on the gene expression profile and corresponding biological behavior of HNSC. Furthermore, even the endocrine milieu of the host was proved to be influential in modulating the progression of HNSC. Gene expression profiling techniques combined with proteomics could help to define and select usefull genetic and biomarkers of progression of HNSC, some of them could well be potential novel therapeutic target. PMID- 15785875 TI - Immunobiology of head and neck cancer. AB - The development of head and neck cancer (HNC) is strongly influenced by the host immune system. Recent evidence for the presence of functional defects and for apoptosis of tumor-infiltrating as well as circulating T cells in patients with HNC emphasizes the fact that their antitumor responses are compromised. It appears that H&N tumors not only effectively escape from the host immune system but also actively corrupt the host antitumor response via several distinct mechanisms. Strategies for restoring immune competence of patients with HNC and for preventing tumor escape are necessary for more effective control of tumor progression. New immunotherapeutic approaches include cytokine-based and DC-based vaccines which are aimed at the restoration of tumor-specific responses and at the protection of immune cells from tumor-induced apoptosis. PMID- 15785877 TI - Molecular therapy of head and neck cancer. AB - Aberrant expression of growth factor receptor systems and dysregulation of the downstream cell signalling molecules have been reported in a wide range of epithelial tumours including head and neck cancer. In some cases, such alterations have been associated with a poor prognosis. In the past 25 years, several antigen specific monoclonal antibodies (mAbs, mouse, chimeric, humanized and human versions), and small molecule kinase inhibitors have been developed that are at different stages of preclinical and clinical developments. Some of these agents (e.g. Herceptin, Iressa, cetuximab, avastin) have already been approved for the treatment of epithelial tumours and may also have potential in the treatment of head and neck cancer patients. This review discusses, the development and potential of these antigen specific agents, in particular the human epidermal growth factor receptor (EGFR) inhibitors, either as a single agent or in combination with other EGFR inhibitors, biological agents (e.g. inhibitors of cycloogenase-2, angiogenesis, insulin like growth factor-I receptor and others), and conventional forms of therapy in the prevention and treatment of head and neck cancer. From preclinical and clinical studies with some of these compounds, it is evident that further detailed studies of biopsies from cancer patients are needed in order to identify markers that can be used not only in the selection of the specific population of cancer patients who would benefit from such antigen specific therapeutic strategies, but also those factors which are responsible for the poor response and the development of a phenotype resistance to such inhibitors. The results of such studies could in turn facilitate the widespread use of such agents in the treatment of a wide range of human cancers including head and neck cancer. PMID- 15785878 TI - Gene therapy for head and neck cancer. AB - The prognosis of patients with advanced head and neck cancer has not changed significantly in the last twenty years, despite concerted efforts to optimize treatment using conventional modalities such as surgery, radiotherapy and chemotherapy. Novel therapeutic approaches based on our increasing understanding of the molecular changes that underlie the development of cancer have the potential to alter this situation. Gene therapy involves the delivery of genetic sequences in to tumour or normal cells for a therapeutic purpose. A number of viral and non-viral vectors have been developed that have the ability to deliver therapeutic genes specifically to tumours. These therapeutic genes can exert their effects by correcting existing genetic abnormalities, by killing cells directly or indirectly through recruitment of the immune system. In this review, the various gene therapy strategies that are under development are presented with particular reference to the treatment of head and neck cancer. PMID- 15785879 TI - The role of B-RAF in melanoma. AB - Melanoma is a form of skin cancer that has a poor prognosis and which is on the rise in Western populations. If detected early, it is easily treated by surgical excision. However, once melanoma metastasises it is notoriously resistant to existing therapies and for many patients the outlook is dismal. Thus a full description of melanoma etiology and a full understanding of the genetic lesions that underlie this disease is required to allow us to develop new and effective therapeutic strategies for its treatment. RAF proteins are a family of serine/threonine-specific protein kinases that form part of a signalling module that regulates cell proliferation, differentiation and survival. In mammals there are three isoforms, A-RAF, B-RAF and C-RAF, and recently it was shown that the B RAF isoform is mutated in a high proportion of melanomas. In light of these exciting findings, we review what we have learned about B-RAF and its role in cutaneous melanoma. PMID- 15785880 TI - Surgery for locally recurrent rectal cancer. AB - PURPOSE: Resection of locally recurrent rectal cancer after curative resection represents a difficult clinical problem and a surgical challenge. The aim of this study was to assess the outcome of a series of patients who underwent resection of locally recurrent rectal cancer with curative intent. METHODS: A retrospective review was performed of 64 patients who underwent surgical exploration with a view to cure for locally recurrent rectal cancer under the care of one surgeon between April 1997 and April 2004. Details were obtained on the primary tumor and the operation, the indication for investigation of recurrence, preoperative imaging, operative findings, morbidity and mortality, and histopathology. RESULTS: The median time interval between resection of primary tumor and surgery for locally recurrent disease was 31 (interquartile range, 21 to 48) months. Twenty-three patients had central disease, 10 patients had sacral involvement, 21 patients had pelvic sidewall involvement, and 10 patients had both sacral and sidewall involvement. Fifty-seven patients underwent resection of the tumor. Thirty-nine of the 57 patients underwent wide resection (abdominoperineal excision of rectum, anterior resection, or Hartmann's procedure) whereas 18 patients (31.6 percent) required radical resection (pelvic exenteration or sacrectomy). Curative, negative resection margins were obtained in 21 of 57 patients who had tumor excision (36.8 percent). Perioperative mortality was 1.6 percent. Significant postoperative morbidity occurred in 40 percent of patients. CONCLUSIONS: This study has shown that a significant proportion of patients with locally recurrent rectal cancer can undergo resection with negative margins. PMID- 15785881 TI - Diagnosis-related group assignment in laparoscopic and open colectomy: financial implications for payer and provider. AB - PURPOSE: In carefully matched patients, the length of hospital stay after laparoscopic colectomy is shorter than after open surgery. Higher operating room costs for laparoscopic surgery are offset by lower costs for hospitalization because of less utilization of pharmacy, laboratory, and nursing services. Clinical outcome is comparable. We examined the effect of the surgical approach for colectomy (open vs. laparoscopic) regarding the reasons for disease-related group assignment to disease-related group 148, and institutional cost under Part A of the U.S. Medicare system. METHODS: Colectomy patients were assigned to either disease-related group 148 (colorectal resection with complications) or disease-related group 149 (colorectal resection without complications) with significant institutional reimbursement implications (disease-related group 149, US 8,310 dollars; disease-related group 148, US 20,291 dollars). A total of 100 consecutive disease-related group 148 patients undergoing laparoscopic colectomy from July 2000 to September 2002 were identified from a prospective database and case-matched with 100 patients undergoing open colectomy. Patients were matched for gender, age, operative procedure, and pathology. A certified coder determined the reason(s) for disease-related group 148 assignment, which were grouped into: preoperative comorbidity, a combination of preoperative comorbidity/postoperative complications, or postoperative complications alone. RESULTS: Significantly more lapararoscopy patients were assigned to disease-related group 148 solely because of preoperative comorbidities (62 percent vs. 21 percent; P < 0.0001). Significantly more patients in the open surgery group were classified as disease related group 148 solely because of postoperative complications (22 percent vs. 42 percent; P < 0.0001). An additional group of patients were assigned to the disease-related group 148 category based on a combination of preoperative and postoperative diagnoses (16 percent vs. 37 percent). The mean direct hospital costs were significantly less for laparoscopy patients (US 3971 dollars vs. US 5997 dollars; P = 0.0095). Increased cost to Part A of Medicare for 20 open surgery patients who "migrated" to disease-related group 148 because of postoperative complications was US 239,620 dollars. CONCLUSIONS: Our data are the first to demonstrate that disease related group assignment can change solely because of a differential rate of postoperative complications for two competing operative techniques. This change occurred at twice the rate for open colectomy and resulted in significantly increased cost to the insurer under a prospective payment program. The savings to the institution coupled with the shortened length of stay offset the potential loss in revenue to the institution. PMID- 15785882 TI - Long-term outcome of surgically treated Crohn's colitis: a prospective study. AB - PURPOSE: Although bowel-sparing techniques have been published for treatment of Crohn's disease of the small bowel because of its relentless nature, extent of resection in Crohn's colitis is still a topic of debate. This study was designed to prospectively evaluate the long-term outcomes of patients with isolated Crohn's colitis to identify patients that may benefit from initial more aggressive resection. METHODS: We identified 179 patients with Crohn's disease operated on for primary colonic disease. They were divided into segmental colectomy, total abdominal colectomy, and total proctocolectomy groups, based on their initial operation. They were further characterized by extent and location of colonic involvement. Long-term outcome variables evaluated included colonic and small-bowel surgical recurrences, postoperative complications and long-term sequelae, long-term need for medical therapy, and need for permanent fecal diversion. RESULTS: Fifty-five patients underwent segmental colectomy, 49 total abdominal colectomy, and 75 total proctocolectomy. Patients with diffuse colonic involvement were significantly less likely to undergo segmental colectomy than total abdominal colectomy (P < 0.0001) or total proctocolectomy (P < 0.0001). Patients with distal involvement or pancolitis were significantly less likely to undergo segmental colectomy than total abdominal colectomy (P < 0.0001) or total proctocolectomy (P < 0.0001). Overall there were 31 patients (24.4 percent) with surgical Crohn's recurrences during follow-up: 19 (38.8 percent) in the segmental colectomy, 8 (22.9 percent) in the total abdominal colectomy, and 4 (9.3 percent) in the total proctocolectomy group. There was a significant difference in time to recurrence between the three groups by log-rank test (P = 0.017). Segmental colectomy patients had a significantly shorter time to first recurrence than total proctocolectomy patients (P = 0.014). After adjusting for extent of disease, the segmental colectomy group had a significantly greater risk of surgical recurrence than the total proctocolectomy group (P = 0.006). Total proctocolectomy patients were significantly less likely to be still taking medications one year after the index operation than total abdominal colectomy patients (P = 0.003) and segmental colectomy patients (P = 0.0003). During follow up, patients with isolated distal disease were significantly more likely to require a permanent stoma than patients with isolated proximal disease (P = 0.004). CONCLUSIONS: A more aggressive approach should be considered in patients with diffuse and distal Crohn's colitis. Total proctocolectomy in the properly selected patients is associated with low morbidity, lower risk of recurrence, and longer time to recurrence. Patients after total proctocolectomy are more likely to be weaned off all Crohn's-related medications. Long-term rate of permanent fecal diversion is significantly higher in patients with distal disease. PMID- 15785883 TI - Treatment of fistulas-in-ano with fibrin sealant in combination with intra adhesive antibiotics and/or surgical closure of the internal fistula opening. AB - PURPOSE: The treatment of fistulas-in-ano with fibrin sealant injection has been moderately successful. Failures can be caused by persistent infection within the tract or early expulsion of the clot. In an attempt to improve the success rate, we examined three modifications of the sealant procedure: the addition of cefoxitin to the sealant, surgical closure of the primary opening, or both. METHODS: A prospective, randomized, clinical trial was performed in which patients were treated with Tisseel-VH fibrin sealant according to previously published procedures. In addition, patients were randomized to receive intra adhesive cefoxitin, surgical closure of the primary opening, or both modifications. Cefoxitin, 100 mg, was added to the sealant for patients randomized to receive intra-adhesive antibiotics. For the appropriate patients, the primary fistula opening was closed with a 3-0 absorbable suture. If fistulas failed to heal, patients were offered a single retreatment with sealant. RESULTS: Twenty-four patients were treated in the cefoxitin arm, 25 in the closure arm, and 26 in the combined arm. Median duration of fistulas was 12 months. Patients were followed for a mean of 27 months postoperatively. There was no postoperative incontinence or complications related to the sealant itself. Initial healing rates were 21 percent in the cefoxitin arm, 40 percent in the closure arm, and 31 percent in the combined arm (P = 0.35). One of five patients in the cefoxitin arm, one of seven patients in the closure arm, and one of six patients in the combined arm were successfully retreated; final healing rates were 25, 44, and 35 percent respectively (P = 0.38). CONCLUSIONS: Treatment of fistula-in-ano with fibrin sealant with closure of the internal opening was somewhat more successful than sealant with cefoxitin or the combination, however this did not achieve statistical significance. None of the three modifications were more successful than historic controls at our institution treated with sealant alone. Therefore, the addition of intra-adhesive cefoxitin, closure of the internal opening, or both are not recommended modifications of the fibrin sealant procedure. PMID- 15785885 TI - Rectal prolapse and cap polyposis: the missing link. PMID- 15785886 TI - Preoperative radiotherapy improves outcome in recurrent rectal cancer. AB - PURPOSE: When local recurrent rectal cancer is diagnosed without signs of metastases, a potentially curative resection can be performed. This study was designed to compare the results of preoperative radiotherapy followed by surgery with surgery only. METHODS: Between 1985 and 2003, 117 patients with recurrent rectal cancer were prospectively entered in our database. Ninety-two patients were suitable for resection with curative intent. Preoperative radiation with a median dosage of 50 Gy was performed in 59 patients; 33 patients did not receive preoperative radiotherapy. The median age of the patients was respectively 66 and 62 years. RESULTS: The median follow-up of patients alive for the total group was 16 (range, 4-156) months. Tumor characteristics were comparable between the two groups. Complete resections were performed in 64 percent of the patients who received preoperative radiation and 45 percent of the nonirradiated patients. A complete response after radiotherapy was found in 10 percent of the preoperative irradiated patients (n = 6). There were no differences in morbidity and reintervention rate between the two groups. Local control after preoperative radiotherapy was statistically significantly higher after three and five years (P = 0.036). Overall survival and metastases-free survival were not different in both groups. Complete response to preoperative radiotherapy was predictive for an improved survival. CONCLUSIONS: Preoperative radiotherapy for recurrent rectal cancer results in a higher number of complete resections and an improved local control compared with patients treated without radiotherapy. Preoperative radiotherapy should be standard treatment for patients with recurrent rectal cancer. PMID- 15785887 TI - Mental disorder and psychologic distress in patients with ulcerative colitis after ileal pouch-anal anastomosis. AB - PURPOSE: The aim of this study was to determine if ileal pouch-anal anastomosis in patients with ulcerative colitis is a psychologic burden for patients, the frequency of mental disorders, the amount of psychologic distress, and their possible disease-related determinants. These factors were studied in patients with ulcerative colitis after ileal pouch anal anastomosis and were compared with ulcerative colitis patients without ileal pouch-anal anastomosis and the general German population. METHODS: A total of 37 patients with ulcerative colitis after ileal pouch-anal anastomosis (age 46.8 +/- 11.8 years; 35 percent female) and 62 patients with ulcerative colitis without ileal pouch-anal anastomosis (age 44.4 +/- 13.9 years; 37 percent female) completed the following questionnaires: medical and sociodemographic questionnaire of the German Competence Network "Inflammatory Bowel Diseases" and the German version of the Hospital Anxiety and Depression Scale. Disease activity was measured in patients with ileal pouch-anal anastomosis by the Pouch Disease Activity Index and in patients without ileal pouch-anal anastomosis by the German Inflammatory Bowel Disease Activity Index. Psychologic distress was assessed by the subscale scores of the Hospital Anxiety and Depression Scale. A probable mental disorder was identified if a patient scored 11 or higher in at least one subscale of the Hospital Anxiety and Depression Scale. RESULTS: The frequency of a probable psychiatric disorder in patients with ileal pouch-anal anastomosis (16 percent) and without ileal pouch anal anastomosis (23 percent) did not differ from that in the general German population (17 percent). Ulcerative colitis patients with or without ileal pouch anal anastomosis did not differ in the amount of psychologic distress. Ileal pouch-anal anastomosis patients had higher levels of anxiety than the general population (P < 0.01). Regression models of disease-related factors predicting mental disorder and psychologic distress showed no significant results. CONCLUSIONS: Ileal pouch-anal anastomosis neither increases nor decreases the frequency of mental disorders or the amount of psychologic distress in ulcerative colitis patients. PMID- 15785888 TI - Anal incontinence after obstetric sphincter tears: outcome of anatomic primary repairs. AB - PURPOSE: Obstetric sphincter tears lead to anal incontinence in 40 to 60 percent of affected women. Primary repair is usually performed without identifying the internal anal sphincter. Since 1999 digestive surgeons have participated in the primary repair of such tears at our hospital. The intention was to perform separate repair of the internal and external anal sphincter in cases of combined tears to achieve a lower incontinence rate than is usually reported after conventional primary repair. The aim of the present study was to evaluate our results after anatomic primary repair. METHOD: A follow-up study was undertaken after all primary repairs performed in 1999 and 2000. It included anal ultrasonography manometry and an assessment of incontinence (Wexner score). RESULTS: A total of 74 women sustained obstetric sphincter tears during the study period, and 71 (96 percent) were assessed after a median of 27 months (range, 14 39 months). Nine women declined investigation with ultrasonography/manometry. Incontinence was present in 22 women (31 percent), of whom 17 had gas incontinence only. The symptoms were mild (Wexner score 1-2) in 11 women (50 percent). None of 17 women with normal ultrasonography results were incontinent versus 20 of 45 with pathologic ultrasonographic results (P = 0.001). The mean sphincter length, squeeze pressure, and resting pressure were significantly higher in women with Wexner scores of 0-2 vs. women with a score of more than 2. Sphincter length was inversely correlated with the degree of incontinence (P < 0.001). CONCLUSIONS: The incontinence rate after anatomic primary repair is low compared with the last decade's reported results after conventional primary repair. A short anal sphincter after repair is associated with a poorer outcome. PMID- 15785889 TI - Laparoscopically-assisted resection rectopexy for rectal prolapse: ten years' experience. AB - PURPOSE: This study has been undertaken to audit a single-center experience with laparoscopically-assisted resection rectopexy for full-thickness rectal prolapse. The clinical outcomes and long-term results were evaluated. METHODS: The data were prospectively collected for the duration of the operation, time to passage of flatus postoperatively, hospital stay, morbidity, and mortality. For follow up, patients received a questionnaire or were contacted. The data were divided into quartiles over the study period, and the differences in operating time and length of hospital stay were tested using the Kruskal-Wallis test. RESULTS: Between March 1992 and October 2003, a total of 117 patients underwent laparoscopic resection rectopexy for rectal prolapse. The median operating time during the first quartile (representing the early experience) was 180 minutes compared with 110 minutes for the fourth quartile (Kruskal-Wallis test for operating time = 35.523, 3 df, P < 0.0001). Overall morbidity was 9 percent (ten patients), with one death (<1 percent). One patient had a ureteric injury requiring conversion. One minor anastomotic leak occurred, necessitating laparoscopic evacuation of a pelvic abscess. Altogether, 77 patients were available for follow-up. The median follow-up was 62 months. Eighty percent of the patients reported alleviation of their symptoms after the operation. Sixty nine percent of the constipated patients experienced an improvement in bowel frequency. No patient had new or worsening symptoms of constipation after surgery. Two (2.5 percent) patients had full-thickness rectal prolapse recurrence. Mucosal prolapse recurred in 14 (18 percent) patients. Anastomotic dilation was performed for stricture in five (4 percent) patients. CONCLUSIONS: Laparoscopically-assisted resection rectopexy for rectal prolapse provides a favorable functional outcome and low recurrence rate. Shorter operating time is achieved with experience. The minimally invasive technique benefits should be considered when offering rectal prolapse patients a transabdominal approach for repair, and emphasis should now be on advanced training in the laparoscopic approach. PMID- 15785890 TI - Sacral neuromodulation in treatment of fecal incontinence following anterior resection and chemoradiation for rectal cancer. AB - PURPOSE: Fecal incontinence may occur in patients who have undergone anterior resection for rectal cancer without presenting sphincter lesions. Chemoradiation may contribute to disrupting continence mechanisms. Treatment is controversial. Assessment of fecal incontinence in patients who agreed to integrate treatment for rectal cancer and treatment with sacral neuromodulation are reported. METHODS: Fecal incontinence following preoperative chemoradiation and anterior resection for rectal cancer was evaluated in four patients. A good response was observed during the percutaneous sacral nerve evaluation test, and so permanent implant of sacral neuromodulation system was performed. Reevaluation was performed at least two months after implant. RESULTS: After device implantation, the mean fecal incontinence scores decreased, and the mean number of incontinence episodes dropped from 12.0 to 2.5 per week (P < 0.05). Permanent implant resulted in a significant improvement in fecal continence in three patients, and incontinence was slightly reduced in the fourth. Manometric parameters agreed with clinical results: maximum and mean resting tone and the squeeze pressure were normal in three patients and reduced in one. In these same three patients, neorectal sensation parameters increased when the preoperative value was normal or below normal and decreased when the preoperative value was higher than normal, whereas in one patient in whom extremely low values were recorded all of the parameters decreased significantly. CONCLUSIONS: Fecal incontinence following anterior resection and neoadjuvant therapy should be carefully evaluated. If a suspected neurogenic pathogenesis is confirmed, sacral neuromodulation may be proposed. If the test results are positive, permanent implant is advisable. Failure of this approach does not exclude the use of other, more aggressive treatment. PMID- 15785891 TI - Evaluation of tumor cell dissociation as a predictive marker of lymph node metastasis in submucosal invasive colorectal carcinoma. AB - PURPOSE: Tumor cell dissociation-the histologic finding of small solid carcinoma cell clusters and groups of dissociated dedifferentiated carcinoma cells at the invasive front-is related to tumor metastasis and patient prognosis. However, few previous reports have examined tumor cell dissociation in submucosal invasive colorectal carcinoma. We investigated the relation between tumor cell dissociation and lymph node metastasis in submucosal invasive colorectal carcinoma. We also examined immunohistochemical expression of E-cadherin and beta catenin in submucosal invasive colorectal carcinoma. METHODS: Submucosal invasive colorectal carcinoma tissue samples from 20 patients with lymph node metastasis and 100 patients without lymph node metastasis were evaluated. Sections stained with hematoxylin and eosin were evaluated for tumor cell dissociation. Immunohistochemistry was used to determine the expression and cellular distribution of E-cadherin and beta-catenin. RESULTS: Tumor cell dissociation was more frequently identified in submucosal invasive colorectal carcinoma cases with lymph node metastasis than in those without lymph node metastasis (P = 0.0001). Decreased membranous expression of E-cadherin occurred more frequently in submucosal invasive colorectal carcinoma cases with lymph node metastasis than in those without lymph node metastasis (P = 0.025). Nuclear expression of beta catenin tended to be present in submucosal invasive colorectal carcinoma cases with lymph node metastasis (P = 0.063). Decreased membranous expression of E cadherin occurred more frequently in submucosal invasive colorectal carcinoma cases with tumor cell dissociation than in those without tumor cell dissociation (P = 0.0023). CONCLUSIONS: Our results suggest that there is a relation between tumor cell dissociation and lymph node metastasis in submucosal invasive colorectal carcinoma. Tumor cell dissociation formation might be related to abnormal expression patterns of E-cadherin and beta-catenin in submucosal invasive colorectal carcinoma. Tumor cell dissociation and decreased membranous expression of E-cadherin would be important predictive markers for lymph node metastasis in submucosal invasive colorectal carcinoma. PMID- 15785892 TI - Operative salvage for locoregional recurrent colon cancer after curative resection: an analysis of 100 cases. AB - PURPOSE: Locoregional recurrence after resection of colon carcinoma is an uncommon and difficult clinical problem. Outcome data to guide surgical management are limited. This investigation was undertaken to review our experience with surgical resection for patients with locoregional recurrence colon cancer, determine predictors of respectability, and define prognostic factors associated with survival. PATIENTS AND METHODS: A prospective database was queried for patients who had recurrent colon cancer between January 1991 and October 2002. Patients were selected for analysis if they had either isolated resectable locoregional recurrence or concomitant resectable distant disease. Disease-specific survival analysis was performed with the Kaplan-Meier actuarial method, and factors associated with outcome were determined by the log-rank test and Cox regression. RESULTS: During this period of time, 744 patients with recurrent colon cancer were identified and 100 (13.4 percent) underwent exploration with curative intent for potentially resectable locoregional recurrence: 75 with isolated locoregional recurrence, and 25 with locoregional recurrence and resectable distant disease. The median follow-up for survivors was 27 months. Locoregional recurrence was classified into four categories: anastomotic; mesenteric/nodal; retroperitoneal; and peritoneal. Median survival for all patients was 30 months. Fifty-six patients had an R0 resection (including distant sites). Factors associated with prolonged disease-specific survival included R0 resection (P < 0.001); age <60 years (P < 0.01); early stage of primary disease (P = 0.05); and no associated distant disease (P = 0.03). Poor prognostic factors included more than one site of recurrence (P = 0.05) and involvement of the mesentery/nodal basin (P = 0.03). The ability to obtain an R0 resection was the strongest predictor of outcome, and these patients had a median survival of 66 months. CONCLUSION: Salvage surgery for locoregional recurrence colon cancer is appropriate for select patients. Complete resection is critical to long-term survival and is associated with a single site of recurrence, perianastomotic disease, low presalvage carcinembryonic antigen level, and absence of distant disease. PMID- 15785893 TI - A scoring system for the strength of a family history of colorectal cancer. AB - BACKGROUND: Family history of colorectal cancer is associated with an increased risk for the disease, although there are many combinations of family history that are hard to correlate with risk status. A scoring system for family history of colorectal cancer was designed to make risk more readily quantifiable. METHODS: A colonoscopy database was used to test the following points system: each first degree relative with colorectal cancer = 3 points; each second-degree relative with colorectal cancer = 1 point. Families with one or more first-degree relative affected under 50 years of age = an extra 3 points. Families with one or more second-degree relative affected under 50 years of age = an extra 1 point. Families with multiple relatives on the same side of the family = an extra 3 points (first-degree relatives), 1 point (second-degree relatives), or 2 points (first-degree and second-degree relatives). Points were added and categories defined as follows: low risk, 1 to 4 points; medium risk, 5 to 7 points; high risk, 8 to 10 points; very high risk, >10 points. A control group of average-risk patients having screening colonoscopy was used. Categories were compared in number of adenomas, hyperplastic polyps, and cancers. RESULTS: The records of 992 patients were used to test the system. Mean adenomas per patient per group were 0.4 for controls, 1.0 for low risk, 1.0 for medium risk, 1.7 for high risk, and 1.7 for very high risk. Cancers per group were 2 of 196 for controls, 8 of 513 for low risk, 3 of 171 for medium risk, 3 of 84 for high risk, and 1 of 28 for very high risk. The score categories were combined to produce revised risk levels of low (score 1 to 7) and high (>7). Average adenomas per patient in the revised categories were 0.4 (control), 1.0 (low risk), and 1.7 (high risk). The odds ratio of having one to two adenomas was 1.73 (1.19-2.50, 95% confidence limits) in the low-risk group and 2.39 (1.41-4.01) in the high-risk group. Odds ratios for having three or more adenomas were 5.70 (2.44-13.32) in the low-risk group and 10.35 (3.97-26.97) in the high-risk group. CONCLUSION: In the two-category system proposed here of quantifying familial risk of colorectal cancer, patients having less than 8 points were at low risk and those with 8 or more were at high risk. Surveillance and chemoprevention protocols can be designed through use of these risk categories. A scoring system for family history of colorectal cancer can make risk assessment easier and facilitate both collaborative studies and patient triage into appropriate screening programs. PMID- 15785894 TI - Efficacy of topical metronidazole (10 percent) in the treatment of anorectal Crohn's disease. AB - PURPOSE: The aim of this study was to investigate the efficacy of 10 percent topical metronidazole in the treatment of symptomatic anorectal Crohn's disease. METHODS: Patients with symptomatic anorectal Crohn's disease were studied in a prospective, nonblinded, nonrandomized study to evaluate the efficacy of topical 10 percent metronidazole. Perianal Crohn's Disease Activity Index and each component (pain, discharge, induration, sexual dysfunction, and type of pathology) were recorded before treatment and at four weeks. Visual analog pain score (0-10) was recorded before treatment and at weekly intervals for up to four weeks. RESULTS: Fourteen patients were studied prospectively for four weeks. Mean Perianal Crohn's Disease Activity Index decreased from 8.8 +/- 0.9 before treatment to 4.1 +/- 0.7 after four weeks' treatment (P < 0.0001). Visual analog pain scores decreased from 5.4 +/- 0.7 before treatment to 2.4 +/- 0.5 at one week (P < 0.001) and to 1.0 +/- 0.4 after four weeks' treatment (P < 0.0001). Of the Perianal Crohn's Disease Activity Index components, significant decreases were noted in pain and discharge (P < 0.0001) and induration (P < 0.001). CONCLUSION: For patients with perianal Crohn's disease, topical 10 percent metronidazole decreases the Perianal Crohn's Disease Activity Index and anorectal pain. PMID- 15785895 TI - Hartmann's colectomy and reversal in diverticulitis: a population-level assessment. AB - PURPOSE: This study was designed to assess the costs and outcomes of colostomy and colostomy reversal in patients with diverticulitis and examine the impact of such procedures on the health care system. METHODS: We employed a retrospective design and used a Washington State administrative database to identify patients undergoing operations with colostomy (1987-2002) who were followed over time. Descriptive and comparative analysis was performed, focusing on patients with diverticulitis. RESULTS: There were 16,556 patients who underwent colostomy and 5,420 (32.7 percent) were for diverticulitis and its related complications (mean age, 64.8 +/- 15.1 years; 53.2 percent female). In patients with diverticulitis, the rate of colostomy reversal was 56.3 percent (80 percent in patients less than 50 years, and 30 percent in patients over 77 years). The in-hospital mortality rate after colostomy reversal was 0.36 percent, and was 2.6 percent in those over 77 years of age. After colostomy reversal a second stoma was used in 3.4 percent, reoperation was required for bleeding complications in 0.6 percent, and infectious complications were noted in 2 percent. The length of time from colostomy to its reversal was approximately five months (138.1 +/- 164 days; interquartile range, 72-156). The relationship between the length of time from colostomy to reversal was evaluated and the adjusted odds of a second stoma being used at the time of colostomy reversal were 45 percent higher (odds ratio, 1.45; 95 percent confidence interval, 1.22, 1.73) for each increase in time interval (<3, 6-9, 9-12, >12 months). CONCLUSIONS: One-third of all colostomies were related to diverticulitis and only 56 percent were reversed. We identified a higher than expected mortality rate among older patients undergoing colostomy reversal. The impact of colostomy and reversal operations on both patients and the health care system is significant. PMID- 15785897 TI - Controlled lateral sphincterotomy for chronic anal fissure. AB - PURPOSE: This study assessed the usefulness of "controlled" lateral sphincterotomy for chronic anal fissures. METHODS: Of 225 patients with chronic anal fissure, 110 underwent traditional sphincterotomy to the level of the dentate line, and 115 underwent controlled sphincterotomy in three steps according to the degree of anal stenosis. In Step 1, the internal sphincter was divided to the proximal level of the fissure. If the anal canal was still stenosed, the division was extended to the level of the dentate line in Step 2. Step 3 was a bilateral internal sphincterotomy. The anal stenosis was evaluated under anesthesia using a new conical calibrator scaled in 1-mm diameter increments. Forty adults without anorectal disease were examined as controls. In a telephone follow-up, 102 patients in the traditional sphincterotomy group and 106 patients in the controlled sphincterotomy group responded. RESULTS: The normal group measured 34.6 +/- 1.4 mm (mean +/- standard deviation). Confounding effects of age, gender, body weight, and height were not significant. Based on the anal caliber measured in the normal group, anal stenosis is present with values of 31 mm and below (mean--2SD of the control value). Of 115 patients in the controlled sphincterotomy group, 90 (78 percent) underwent sphincterotomy below the level of the dentate line, 18 (16 percent) underwent sphincterotomy to the level of the dentate line, and 7 (6 percent) underwent bilateral sphincterotomy. None had incontinence of feces or leakage of stool. Ten of 102 patients (10 percent) in the traditional sphincterotomy group and 2 of 106 patients (2 percent) in the controlled sphincterotomy group complained of minor incontinence, such as gas incontinence, minor staining, or urgency (P = 0.017). There was one recurrence in the traditional sphincterotomy group. CONCLUSION: Controlled lateral sphincterotomy could be a way of overcoming the risk of incontinence with lateral internal sphincterotomy for chronic anal fissure. PMID- 15785896 TI - Preoperative serum vascular endothelial growth factor is not a marker for subsequent recurrence during long-term follow-up of colorectal cancer patients. AB - BACKGROUND: Serum vascular endothelial growth factor has been associated with stage of disease in colorectal cancer patients. We investigated whether preoperative serum vascular endothelial growth factor can provide any relevant clinical prognostic information during long-term follow-up of colorectal cancer patients. METHODS: Preoperative serum samples of 79 colorectal cancer patients and serum of 28 healthy controls were stored at -80 degrees C until later vascular endothelial growth factor analysis by enzyme-linked immunosorbent assay technique and carcinoembryogenic antigen concentration measurement were performed. There were three patient groups for comparison: 21 patients with overt liver metastases, 18 patients who developed recurrent disease after initial curative surgery, and 40 patients who remained disease-free for at least five years. RESULTS: We could not demonstrate any significant difference in serum vascular endothelial growth factor values between the patient groups and controls, nor between the three patient groups (Mann-Whitney U test). There was no relevant correlation between serum vascular endothelial growth factor and carcinoembryogenic antigen concentrations (Pearson r = 0.2; P = 0.07). CONCLUSION: Although vascular endothelial growth factor has been shown in previous studies to be a potent inducer of angiogenesis and metastases formation, the present data demonstrate that preoperative serum vascular endothelial growth factor concentration does not provide any relevant individual prognostic information in patients with colorectal cancer. PMID- 15785898 TI - Intraperitoneal cytokine productions and their relationship to peritoneal sepsis and systemic inflammatory markers in patients with inflammatory bowel disease. AB - PURPOSE: This prospective, blinded study was designed to assess intraperitoneal cytokine productions and their relationship to clinical presentations and systemic inflammatory markers in patients with inflammatory bowel disease. METHODS: Fifty patients who required abdominal surgery for active inflammatory bowel disease (ulcerative colitis, 27; and Crohn's disease, 23) were investigated. Interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha levels in intraperitoneal fluid (obtained by intraoperative lavage) and plasma were measured by enzyme-linked immunosorbent assay. To compare intraperitoneal cytokine productions between diseases with a different degree of inflammation, intraperitoneal cytokine measurement also was performed for patients who required surgery for colorectal cancer (n = 25) and acute appendicitis (n = 25). RESULTS: The median intraperitoneal cytokine (interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha) levels were significantly higher in patients with inflammatory bowel disease than in patients with colorectal cancer and acute appendicitis. In patients with inflammatory bowel disease, intraperitoneal cytokine levels were significantly higher in patients with intraperitoneal sepsis (abscess/fistula) than in patients without intraperitoneal sepsis at laparotomy. Intraperitoneal cytokine levels did not correlate with type of disease (ulcerative colitis/Crohn's disease), age, gender, duration of disease before surgery, preoperative medical treatment, and the extent or site of disease. There were no significant correlations between intraperitoneal cytokine levels and the following systemic inflammatory markers: plasma cytokine levels, white blood cell count, platelet count, erythrocyte sedimentation rate, and C-reactive protein levels. Postoperative intraperitoneal septic complications (anastomotic leak/abscess/enterocutaneous fistula) more frequently occurred in patients with higher intraperitoneal cytokine levels. CONCLUSIONS: Intraperitoneal cytokine productions were greatly elevated in patients with inflammatory bowel disease, and their levels correlated with the presence of intraperitoneal sepsis at laparotomy and development of postoperative intraperitoneal septic complications. Intraperitoneal cytokine levels showed no correlations with systemic inflammatory markers. PMID- 15785899 TI - Terminal ileal stricture in Crohn's disease: treatment using a metallic enteral endoprosthesis. AB - Enteral strictures are a frequent indication for surgery in Crohn's disease. Postoperative complications are increased in patients with poor preoperative nutritional status, which is common in this patient population. We present a 49 year-old female with longstanding Crohn's disease admitted to our Digestive Health Center with four weeks of increasing abdominal symptoms and radiographic evidence of small-bowel obstruction caused by ileal stricture. Given her poor nutritional status, our team elected to pursue metallic enteral stenting as a bridge to surgical resection. Two Wallstents were placed; luminal patency was subsequently confirmed by a fluoroscopic study. The patient tolerated regular diet and was discharged. When seen in follow-up, she remained asymptomatic and wished to defer surgical intervention indefinitely. PMID- 15785900 TI - Quality of life, functional outcome, and complications of coloplasty pouch after low anterior resection. AB - PURPOSE: The colonic J-pouch has been used to improve bowel function in patients undergoing low colorectal or coloanal anastomosis. However, a narrow pelvis, difficulties in reach, a long anal canal with prominent sphincters, or a fatty mesentery may turn this technique into a technically challenging procedure in certain patients. In these circumstances, "coloplasty" offers an alternative to a straight anastomosis. The purpose of this study was to compare the quality of life, functional outcome, and complications between patients undergoing coloplasty, colonic J-pouch, or straight anastomosis. METHODS: Altogether, 162 patients who underwent coloanal or low colorectal anastomosis between 1998 and 2001 were studied. Data collected included demographics, length of follow-up, technique and type of anastomosis, complications, quality of life, and functional outcome. Results were analyzed according to use of a coloplasty (n = 69), colonic J-pouch (n = 43), or straight anastomosis (n = 50). The choice of the technique was based on the surgeon's preference. Usually coloplasty or straight anastomosis was favored in male patients with a narrow pelvis or when a handsewn anastomosis was used. RESULTS: Quality of life assessment with the short form-36 questionnaire revealed better scores in coloplasty and colonic J-pouch groups. The coloplasty (1.0 +/- 1.7) and colonic J-pouch (1.0 +/- 1.2) groups had fewer night bowel movements than the straight anastomosis group (1.5 +/- 2.0) (P < 0.05). The coloplasty group also had fewer bowel movements per day than the straight anastomosis group (3.8 +/- 2.9 vs. 4.8 +/- 3.6; P < 0.05); also, less clustering and less antidiarrheal medication use were observed than in the straight anastomosis group. Colonic J-pouch patients with handsewn anastomosis had a higher anastomotic leak rate (44 percent) than the patients in the coloplasty with handsewn anastomosis group (3.6 percent). CONCLUSIONS: Coloplasty seems to be a safe, effective technique for improving the outcome of low colorectal or coloanal anastomosis. It is especially applicable when a colonic J pouch anastomosis is technically difficult. PMID- 15785901 TI - Stapled hemorrhoidopexy vs. diathermy excision for fourth-degree hemorrhoids: a randomized, clinical trial and review of the literature. AB - PURPOSE: The aim of this prospective study was to compare the results of stapled hemorrhoidopexy with those of conventional diathermy excision for controlling symptoms in patients with fourth-degree hemorrhoids. METHODS: Thirty-one patients with symptomatic, prolapsed irreducible piles were randomized to either stapled hemorrhoidopexy (n = 15) or diathermy excision (n = 16). The primary outcome measure was the control of hemorrhoidal symptoms one year after operation. RESULTS: The two procedures were comparable in terms of pain relief and disappearance of bleeding. Recurrent prolapse starting from the fourth month after operation was confirmed in 8 of 15 patients in the stapled group and in none in the diathermy excision group: two-tailed Fisher's exact test P = 0.002, RR 0.33, 95 percent confidence interval 0.19-0.59). Five of these patients responded well to a later conventional diathermy hemorrhoidectomy. Persistence of itching was reported in six patients in the stapled group and in one of the diathermy excision group (P = 0.03). On the other hand, six patients in the stapled group and none in the diathermy excision group experienced tenesmus (P = 0.007). CONCLUSIONS: Stapled hemorrhoidopexy was not effective as a definitive cure for the symptoms of prolapse and itching in patients with fourth-degree hemorrhoids. Moreover, stapled hemorrhoidopexy induced the appearance of a new symptom, tenesmus, in 40 percent of the patients. Therefore conventional diathermy hemorrhoidectomy should continue to be recommended in patients with symptomatic, prolapsed, irreducible piles. PMID- 15785902 TI - Laparoscopic or transanal repair of rectocele? A retrospective matched cohort study. AB - PURPOSE: The aim of the study was to analyze the functional and physiologic outcome of patients undergoing laparoscopic rectocele repair compared to a matched cohort undergoing transanal repair. METHODS: Forty patients with a rectocele who had undergone laparoscopic pelvic floor repair by a laparoscopic gynecologist were matched for age and rectocele size with 40 patients who had undergone a transanal repair by a colorectal surgeon. All patients had clinical evidence of a symptomatic rectocele. All patients were assessed postoperatively with a quality of life (SF-36) score, a modified St. Mark's continence score, a urinary dysfunction score, a Watt's sexual dysfunction score, and a linear analog patient satisfaction score. Fifteen patients in each group had also undergone preoperative and postoperative anal manometry. RESULTS: At 44 months median follow-up, the transanal approach resulted in significantly more patients reporting bowel symptom alleviation (P < 0.002) and higher patient satisfaction (P < 0.003). The bowel symptom improvement was also sustained over a significantly longer period (P < 0.03). Only 11 patients (28 percent) in the laparoscopic group reported more than 50 percent improvement in their bowel symptoms compared to 25 patients (63 percent) in the transanal group. On univariate analysis of 50 percent bowel symptom improvement, a larger rectocele (P < 0.009), transanal repair (P < 0.02), and presenting with obstructive defecation rather than fecal incontinence (P < 0.03) were statistically significant. Rectocele size (P < 0.012) and treatment cohort (P < 0.006) remained significant on multivariate analysis. Postoperatively, bowel symptom alleviation correlated with patient satisfaction in both groups (P < 0.015). Although not statistically significant, five patients (13 percent) in the transanal group developed postoperative fecal incontinence, which was associated with a low maximum anal resting pressure preoperatively that was further diminished postoperatively (P > 0.06). Only one patient (3 percent) in the laparoscopic group reported a decline in fecal continence, but four patients (10 percent) reported worsening of their symptoms of obstructed defecation. Postoperative dyspareunia was reported by 24 patients in total (30 percent), with significantly more in the transanal group (P > 0.05). CONCLUSIONS: The transanal repair results in a statistically greater alleviation of bowel symptoms and greater patient satisfaction scores. However, this approach may have a greater degree of functional co-morbidity than the laparoscopic rectocele repair. PMID- 15785903 TI - Rectal intussusception: characterization of symptomatology. AB - PURPOSE: Rectal intussusception is a common finding at evacuation proctography; however, its significance has been debated. This study was designed to characterize clinically and physiologically a large group of patients with rectal intussusception and test the hypothesis that certain symptoms are predictive of this finding on evacuation proctography. METHODS: A total of 896 patients underwent evacuation proctography from which three groups were identified: those with isolated rectal intussusception (n = 125), those with isolated rectocele (n = 100), and those with both abnormalities (n = 152). Multivariate analyses were used to identify symptoms predictive of findings by evacuation proctography. RESULTS: The symptoms of anorectal pain and prolapse were highly predictive of the finding of isolated intussusception over rectocele (odds ratio, 3.6, P = 0.006; odds ratio, 4.9, P < 0.001) or combined intussusception and rectocele (odds ratio, 2.9, P = 0.02; odds ratio, 2.4, P = 0.03). The symptom of "toilet revisiting" was associated with the finding of rectoanal intussusception (odds ratio, 3.55, P = 0.04). Although patients with mechanically obstructing intussuscepta evacuated slower and less completely (P < 0.001) than those with nonobstructing intussuscepta, no symptom was predictive of this finding on evacuation proctography. CONCLUSIONS: Although certain symptoms are predictive of the finding of rectal intussusception, there is a wide overlap with symptoms of rectocele, another common cause of evacuatory dysfunction. Furthermore, the observation that "obstruction to evacuation" made on proctography had no impact on the incidence of evacuatory symptoms suggests that beyond simply demonstrating the presence of an intussusception, analysis of proctography and subclassifying intussusception morphology seems of little clinical significance, and selection for surgical intervention on the basis of proctographic findings may be illogical. PMID- 15785905 TI - The rural surgeon. A new forum on the World Journal of Surgery. PMID- 15785907 TI - Gastric cancer by multidetector row CT: preoperative staging. AB - The role of computed tomography (CT) in the preoperative staging of gastric cancer, even if controversial, may be fundamental for evaluating the local extent and nodal involvement of the disease, especially in locally advanced cases. However, previous results of CT staging have not been satisfactory for predicting the invasive depth of the tumor or possible nodal metastases. Recently introduced multidetector row CT (MDCT) and three-dimensional (3D) imaging are expected to overcome the limitations in cancer staging by offering rapid and accurate information for space perception, detailed hemodynamics, and real-time 3D processing of volumetric data sets. In particular, virtual endoscopic imaging may be helpful for detecting early gastric cancer. In our experience, T and N stagings of gastric cancer were improved with 3D MDCT when using volume rendering and virtual endoscopic imaging compared with conventional axial two-dimensional (2D) CT (accuracy of T staging with 3D vs. 2D CT images, 84% vs. 77%; accuracy of N staging, 63% vs. 61% with 3D vs. 2D images, respectively). In particular, the detection rate of early gastric cancer was markedly increased up to 96% when using 3D MDCT. Therefore, we believe that 3D MDCT of the stomach may enhance the performance of CT in the preoperative evaluation of patients who have gastric cancer by offering easy early detection of lesions and accurate tumor staging through the 3D imaging process. PMID- 15785908 TI - Primary hepatic carcinoid tumor: dynamic CT findings. AB - Most carcinoid tumors involve the gastrointestinal tract or respiratory system. Primary hepatic carcinoid tumor is a rare entity. A 46-year-old woman presented with abdominal pain, vomiting, and diarrhea. She was diagnosed with primary hepatic carcinoid tumor based on radiologic and laboratory findings, namely a mass in the fifth segment of the liver and markedly elevated levels of 5 hydroxyindole acetic acid in the urine. Histologic and immunohistochemical findings of the resected liver segment showed a malignant carcinoid tumor of the liver. This case is of interest because of the rarity of this neoplasm. This report describes the case and reviews the clinical features, radiologic findings, and treatment in previously reported cases. PMID- 15785909 TI - [Rotational stability in intraocular lenses with C-loop haptics versus Z haptics in cataract surgery. A prospective randomised comparison]. AB - The aim of the study was to compare the rotational stability of intraocular lenses (IOLs) with C-loop haptics and those with Z haptics. A total of 50 patients with cataracts were prospectively randomised in equal numbers using a design equivalent to IOL MS 612 S (C-loop haptic) and MS 6120 (Z haptic, both HumanOptics, Erlangen). Complete ophthalmological examinations were performed including assessment of the IOL rotation after 1 day, 1 month and 3 months postoperatively. The significances between the groups were evaluated using the Wilcoxon test. The Fisher exact test was used for the primary finding of the study (rotation of at least 10 degrees ). The IOL group with the C-loop haptics showed a median lens rotation of 0 degrees on the first day and 2 degrees clockwise after 1 month and 3 months. In the Z haptic group, there was no median rotation in the IOL group. Moreover, the range of IOL rotation of the C-loop haptics was broader (3 months postoperatively: maximum in the C-loop haptics: 21 degrees with 15.5 degrees in the Z haptics). There was no significant difference at any time. At 3 months postoperatively, 32% of the C-loop haptic IOL and 16% of the Z haptic IOL rotated at least 10 degrees (P=0.32). In 59% of the IOL with C loop haptics, the direction of the rotation was clockwise. This was 40% for the IOL with Z haptics (P=0.33). At 3 months postoperatively, both IOLs demonstrated good rotational stability with a low mean deviation from the target axis. IOLs with Z haptics showed a tendency to greater rotational stability than C-loop haptics but without any significant difference. There was no significant difference in visual rehabilitation. PMID- 15785911 TI - [Laser in situ keratomileusis (LASIK) and scanning laser ophthalmoscopy]. AB - BACKGROUND: To determine the influence of Laser in situ keratomileusis (LASIK) on the measurements of retinal thickness and optic nerve head topography using the Retinal Thickness Analyzer (RTA). METHODS: RTA measurements were performed before and after LASIK. Forty-eight eyes of 25 healthy subjects were included. Mean age was 40.0+/-10.6 years. Mean preoperative refractive error (spherical equivalent) was -3.3+/-3.6 dpt, and 0.2+/-0.9dpt postoperatively. Correlation between ablation depth and duration and change of retinal thickness postoperatively was performed. All patients received a pachymetry preoperatively. RESULTS: Postoperatively, mean retinal nerve fiber layer thickness (MRNFL) and cross sectional area (RNFL cross section area) significantly decreased after LASIK (MRNFL preop: 0.18 mm, postop: 0.11 mm, P =0.026, RNFL cross section preop: 1.17 mm(2), postop: 0.71 mm(2), P =0.015). Ablation depth revealed a significant correlation with changes in retinal thickness measurements postoperatively (Delta MRNFL: Ablation depth, P =0.001, r=-0.5). Duration of the ablation was not significantly correlated to the MRNFL measurements postoperatively (P =0.27, r= 0.08). No correlation was found between the central corneal thickness and the change in retinal thickness after LASIK (P =0.51). CONCLUSION: Due to changes in corneal architecture after LASIK, measurements using RTA reveal a decrease in MRNFL and RNFL cross section area. These changes are likely artifacts. Further studies with a longer follow-up are desirable. PMID- 15785910 TI - [Esterase activity of human organotypic cornea construct (HCC) as in vitro model for permeation studies]. AB - Organotypic cornea equivalents are used as in vitro models for permeation studies. Many ophthalmic drugs are applied as ester prodrugs to achieve a higher bioavailability. The esterase activity of three corneal human cell lines (epithelial, stromal, endothelial cells) as well as of excised porcine cornea, human donor cornea and human cornea construct (HCC) was investigated and compared. Esterase activity was determined using p-nitrophenyl acetate and hydrocortisone acetate (HCA) as esterase substrates. Hydrocortisone acetate permeation across porcine cornea, human donor cornea and HCC was studied in vitro using Franz-diffusion cells. Corneal epithelial cells showed the highest esterase activity and only small differences to keratocytes and endothelial cells were detectable. The permeation barrier properties of the different corneal tissues were very similar in the case of HCA permeation whereas HCA metabolism rates were in the ranking order of porcine cornea > HCC > human donor cornea. Permeation and metabolism studies indicate that the in vitro permeation model HCC is able to adequately convert hydrocortisone acetate to hydrocortisone. PMID- 15785912 TI - [The effect of brimonidine on the pupillary reflex. A pupillographic study in healthy volunteers]. AB - PURPOSE: The effect of brimonidine in comparison with acetazolamide on pupillary reflex was investigated in 18 volunteers. METHODS: Infrared pupillography was performed with white diode light of 200 ms duration to measure pupil diameter, constriction latency, reaction time, constriction amplitude, and relative constriction amplitude. The measurements were performed according to a fixed schedule including a phase without medication to determine the baseline level. Data were analyzed by Student's paired t-test. RESULTS: Application of brimonidine and acetazolamide led to a significantly reduced intraocular pressure as well as static and dynamic differences in the pupillary reflex. The pupil diameter measurements were significantly smaller after both medications in comparison to baseline. The reduction of pupil diameter after brimonidine led to significantly reduced contraction amplitude and prolonged latency. CONCLUSION: Application of brimonidine leads to significant miosis, which might due to the affinity to alpha(2)-receptors with reduction of noradrenaline release in the synapse. This effect may play a role in a higher decrease of intraocular pressure by increasing aqueous humor outflow in comparison to clonidine and apraclonidine, but further investigations are required. PMID- 15785914 TI - Patient follow-up: a call for organized "research". PMID- 15785915 TI - Signal failure of type 2 comprehensive unattended sleep studies in patients with suspected respiratory sleep disordered breathing. AB - We designed this study to assess the signal failure and sensor loss of unattended type 2 comprehensive polysomnography (PSG) and compared that with in-lab attended PSG. Type 2 PSG was performed for 41 patients. The signal failure was estimated and compared to the signal failure in 60 patients for the in-lab PSGs. The signal failure in each individual electroencephalographic (EEG) channel, complete EEG signals, electro-oculography (EOG), naso-oral flow, and thoracic belt were significantly greater in the unattended sleep studies. The failure rate for the different signals ranged from 0.128 min in electrocardiography (EKG) to 67.36 min in the thoracic belt signal. However, that did not affect the success rate of the studies. Acceptable scorable data was available in 97% of the performed unattended PSGs. Unattended type 2 sleep studies can be performed for clinical use in the evaluation of sleep disordered breathing with low signal failure and sensor loss if the proper hook-up procedure was followed. PMID- 15785916 TI - Effect of CPAP treatment on inspiratory arousal threshold during NREM sleep in OSAS. AB - The maximal inspiratory effort recorded at the end of apnea has been considered as an index of arousal threshold in obstructive sleep apnea syndrome (OSAS). Previous investigations have shown that the arousal threshold is higher in patients with OSAS than in normal subjects. The aim of the present study was to investigate the effect of continuous positive airway pressure (CPAP) treatment on the inspiratory-effort-related arousal threshold in patients with OSAS. In ten male patients, 40 episodes of apnea during stage 2 non-REM (NREM) sleep were analyzed. Apnea duration (t), esophageal pressure (Pes) at the first occluded breath (Pes1), the minimum of the three initial Pes swings (Pes min), the maximum of the three final Pes swings (Pes Max), DeltaPes (Pes Max-Pes min), RPes (rate of increase of intrathoracic pressure, DeltaPes/t), n (number of occluded breaths during apnea), DeltaPes/n, n/t, and SaO(2) were determined before and after occlusion. These apneic episodes were compared to ten episodes of apnea provoked by a mask occlusion device after 1, 7, 30, and 90 days of CPAP treatment. The therapy resulted in a decrease in the inspiratory-effort-related arousal threshold, as measured by a reduction of Pes Max, without significant changes in apnea duration and apnea-related hypoxemia. Pes1 and DeltaPes/n, which are markers of respiratory drive, significantly decreased between observations. CPAP treatment decreases the inspiratory-effort-related arousal threshold and induces a decrease in ventilatory drive in response to upper airway occlusion. PMID- 15785917 TI - Prospective evaluation of an oral appliance in the treatment of obstructive sleep apnea syndrome. AB - The purpose of this study was to investigate the effects of an oral appliance (OA), with and without mandible advance, in the treatment of obstructive sleep apnea syndrome (OSA). Twenty-four patients diagnosed with OSA agreed to participate in this study. The patients were treated for 3 months (with a removable soft elastic silicone positioner customized with thermoplastic silicone and with a 5-mm opening). Patients were selected, using a randomized design, to receive an OA model either with (12 patients) or without advance (12 patients). Before treatment, a snoring questionnaire, the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36), the Functional Outcomes of Sleep Questionnaire (FOSQ), the Epworth Sleepiness Scale (ESS), and polysomnography were completed. Fifteen subjects completed the protocol (13 men, two women). With respect to basal values, the mandible-advanced OA group presented a decrease in the mean apnea-hypopnea index (AHI) (33.8+/-4.7 versus 9.6+/-2.1; p<0.01), number of arousals per hour (33.8+/-13.9 versus 16.0+/-1.5; p<0.05), ESS score (14.7+/-5.1 versus 5.1+/-1.9; p<0.05), snoring score (15.4+/-1.9 versus 10.1+/-3.2; p<0.05), and total FOSQ score (78.1+/-22.6 versus 99.3+/-14.4; p<0.05). After treatment, the non-advanced group presented a decrease in the mean AHI (24.0+/-12.2 versus. 11.7+/-7.9; p<0.05). However, no significant differences were found in the number of arousals per hour, ESS score, snoring, and total FOSQ score in the non advanced group. Neither study group showed significant difference in mean SF36 scores. Oral appliances, especially those that advance the mandible, offer an effective treatment for OSA. PMID- 15785918 TI - The use of a predicted CPAP equation improves CPAP titration success. AB - Titration of continuous positive airway pressure (CPAP) is performed to determine the CPAP setting to prescribe for an individual patient. A prediction equation has been published that could be used to improve the success rate of CPAP titrations. The goals of this study were: (1) to test the hypothesis that the use of the prediction equation would achieve a higher rate of successful CPAP titrations; (2) to validate the equation as an accurate predictor of the prescribed CPAP setting and determine the factors that influence the accuracy of the prediction equation. A total of 224 patients underwent CPAP titration prior to using the equation, with a starting pressure of 5 cm H(2)O. A total of 192 patients underwent CPAP titration using the equation-predicted CPAP level as the starting pressure (median starting pressure of 8 cm H(2)O [interquartile range 7, 10 cm H(2)O]). The percentage of successful studies, as defined by a 50% decrease in the apnea-hypopnea index (AHI) and a final AHI < or =10 cm H(2)O, increased from 50% to 68% (p<0.001), while the number of patients who were prescribed a CPAP level that had not been tested decreased from 22% to 5% (p<0.001). The equation was not accurate in predicting the prescribed level of CPAP, with only 30.8% of the patients with a prescribed pressure < or =3 cm H(2)O of the predicted pressure. Female gender was the only predictor of a prescribed pressure < or =3 cm H(2)O from the predicted pressure (odds ratio 3.45, 95% confidence intervals 1.67, 7.13, p<0.001). A CPAP prediction equation modestly increases the rate of successful CPAP titrations by increasing the starting pressure of the titration. The equation does not accurately predict the prescribed CPAP level, reaffirming the need for a titration study to determine the optimal prescribed level in a given patient. PMID- 15785919 TI - FEF(25-75)/FVC measurements and extrathoracic airway obstruction in obstructive sleep apnea patients. AB - The aims of this study were to evaluate patients with obstructive sleep apnea syndrome (OSAS) with regards to dysanapsis (airway size relative to lung size) and to demonstrate the differences between the patients with and without extrathoracic airway obstruction. The study population consisted of 15 patients with OSAS and 14 age and body mass index (BMI) matched control subjects. OSAS patients and control subjects showed similar characteristics in FEV(1), FEV(1)/FVC, FEF(25-75), and FEF(25-75)/FVC ratios. Expiration reserve volume was significantly higher in the control group than in OSAS patients (p<0.01). Six patients exhibited extrathoracic airway obstruction while awake. Of these, three had also a sawtooth pattern in their flow-volume curves. The remaining nine patients had no extrathoracic airway obstruction and had lower apnea-hypopnea indexes (AHI) than the obstruction group (p<0.05). OSAS patients and age- and BMI matched healthy controls had similar characteristics in terms of dysanapsis. In addition, there was no relation between the FEF(25-75)/FVC ratio and AHI, MinO(2), and MeanO(2). Extrathoracic airway obstruction may be a feature of only severe OSAS patients. PMID- 15785920 TI - Sleep apnea avoidance pillow effects on obstructive sleep apnea syndrome and snoring. PMID- 15785921 TI - Report from the first conference of the European Dental Sleep Medicine Academy. PMID- 15785925 TI - Polo-like kinase isoforms in breast cancer: expression patterns and prognostic implications. AB - Polo-like kinase (PLK) family members are known to be functionally involved in mitotic signaling and in cytoskeletal reorganization in both normal and malignant cells. In this study, expression of PLK1 and PLK3 was determined immunohistochemically in tissue specimens of 135 breast carcinomas, and expression was correlated with clinicopathological parameters and patient prognosis. Strong PLK isoform overexpression was observed in 42.2% (PLK1) and 47.4% (PLK3) of breast carcinomas when compared with non-transformed breast tissue. A positive correlation of PLK isoform expression with tumor grade, vascular invasion, erbB2/HER-2 expression and markers of proliferative activity was evident. Additionally, an inverse correlation of PLK isoform expression and estrogen receptor status was observed. Overexpression of PLK3 but not of PLK1 was significantly linked to reduced median overall (P<0.001) and relapse-free (P=0.021) survival time. PLK3 expression remained an independent prognostic factor for overall (RR=3.2, P=0.002) and relapse-free (RR=2.9, P=0.049) survival in multivariate survival analysis. These results suggest PLK3 as a novel independent prognostic marker in breast cancer and hint toward a role for PLK isoform overexpression in disease progression. Therefore, in vivo inhibition of PLK family members might represent a rewarding approach in the development of new anticancer drugs for this tumor entity. PMID- 15785924 TI - Topical brimonidine reduces collateral damage caused by laser photocoagulation for choroidal neovascularization. AB - PURPOSE: To evaluate the neuroprotective efficacy of brimonidine applied topically to the eyes of patients undergoing laser treatment for extrafoveal or juxtafoveal choroidal neovascularization (CNV). METHOD: In this prospective, controlled, double-masked pilot study, 20 eyes were randomized to receive either brimonidine 0.2% (study group, 11 eyes) or placebo (matched control group, 9 eyes). Medications were topically applied twice a day during a period of 4-48 h before laser treatment and were continued for 1 month. RESULTS: Two eyes in each group had severe visual loss owing to recurrence of CNV. In the remaining 16 eyes there was a significant improvement in the study group, but no improvement in the control group (P=0.027). CONCLUSION: Topical brimonidine improves the visual outcome of laser-treated classic extrafoveal or juxtafoveal CNV, possibly by protecting the neuroretina against collateral damage caused by the laser treatment. PMID- 15785926 TI - CD2-associated protein in human urogenital system and in adult kidney tumours. AB - We studied expression of CD2-associated protein (CD2AP) in human urogenital system and in adult kidney tumours. In the cortex of normal kidney, CD2AP was expressed in all types of tubules and in the glomeruli. Labelling was more intense in cytokeratin 7- and in Tamm-Horsfall-positive tubules than in proximal tubules. In the medulla, expression was observed in the collecting ducts. Urothelium and the epithelium of prostatic acini, seminal vesicles, seminiferous tubules, epididymal ducts, Fallopian tube, endometrium and endocervix as well as granulosa cells showed moderate to strong CD2AP positivity. In syncytiotrophoblast, the expression was weaker than in cytotrophoblast. Endometrial stroma was negative, but decidualised stroma was weakly positive. Clear cell renal cell carcinoma (RCC) (n=63) showed a weak expression. Type-I papillary RCCs (n=4) and papillary adenomas (n=3) were negative. The epithelium lining the cysts in multilocular cystic RCCs (n=3) and in cystic nephroma (n=1) was strongly positive. Chromophobe RCCs (n=2), oncocytomas (n=3) and urothelial carcinomas (n=2) were moderately positive. The results show that CD2AP displays a specific expression pattern in human urogenital organs and that distinct expression is shown in several types of kidney tumours but not in type-I papillary RCCs or in papillary adenomas. PMID- 15785927 TI - Benign Mullerian inclusions coexisting with breast metastatic carcinoma in an axillary lymph node. PMID- 15785928 TI - [Reflux and sinusitis]. PMID- 15785930 TI - Obesity and fat quantification in lean tissues using three-point Dixon MR imaging. AB - BACKGROUND: It has been suggested that increased hepatic and intramuscular fat is associated with insulin resistance, and that increased pancreatic fat is related to impaired insulin secretion. OBJECTIVE: We postulated that in obese nondiabetic teenagers insulin levels would be directly related to increases in intramuscular and hepatic fat and inversely related to increases in pancreatic fat. MATERIALS AND METHODS: MRI was used to assess the percentage of fat in the liver, muscle and pancreas in 15 healthy Mexican-American girls, 14-17 years old, with body mass indexes (BMIs) ranging from 17.7 kg/m(2) to 46 kg/m(2). RESULTS: Strong correlations were observed between BMI and fat content in the liver, muscle, and pancreas (r(2)s between 0.50 and 0.89; P<0.003). Serum insulin levels were closely associated with fat measures in the muscle and liver (r(2s)=0.63 and 0.29, and P=0.001 and P=0.023, respectively). In contrast to our hypothesis, fat content in the pancreas was also directly related to insulin secretion (r(2)=0.74; P=0.001). SUMMARY: We conclude that in nondiabetic teenagers, obesity is associated with an increased accumulation of fat in the pancreas without impairment of insulin secretion. PMID- 15785929 TI - Acinetobacter baumannii ventilator-associated pneumonia: epidemiological and clinical findings. AB - OBJECTIVE: To investigate prognostic factors and predictors of Acinetobacter baumannii isolation in ventilator-associated pneumonia (VAP). We specifically analyzed these issues for imipenem-resistant episodes. DESIGN AND SETTING: All episodes of VAP are prospectively included in a database. Information about risk factors was retrieved retrospectively. PATIENTS: Eighty-one patients exhibiting microbiologically documented VAP: 41 by A. baumannii (26 by imipenem-resistant) and 40 by other pathogens. MEASUREMENTS AND RESULTS: The following variables were noted: underlying diseases, severity of illness, duration of mechanical ventilation and of hospitalization before VAP, prior episode of sepsis, previous antibiotic, corticosteroid use, type of nutrition, renal replacement therapy, reintubation, transportation out of the ICU, micro-organisms involved in VAP, concomitant bacteremia, clinical presentation, Sequential Organ Failure Assessment (SOFA) scale on the day of diagnosis, and adequacy of empirical antibiotic therapy. Prior antibiotic use was found to be associated with development of VAP by A. baumannii (OR 14). Prior imipenem exposure was associated with the isolation of imipenem-resistant strains (OR 4). SOFA score on the day of diagnosis was the only predictor of in-hospital mortality (OR 1.22); adequacy of empirical antibiotic therapy was a protective factor (OR 0.067). CONCLUSIONS: Our results confirm that prior exposure to antimicrobials is an independent predictor for the development of A. baumannii VAP, the prognosis of which is similar to that of infections caused by other pathogens. This study highlights the importance of initial antibiotic choice in VAP or whatever cause. PMID- 15785931 TI - Cystic malformations of the neck in children. AB - The most common cystic malformations of the neck are the result of abnormal embryogenesis involving the thyroglossal duct (TGD), lymphatic primordia and the branchial apparatus. When the basic embryology of these structures is considered, a reasonable differential diagnosis-and in some cases a definitive diagnosis-can be achieved based on the location and the imaging characteristics of the cystic mass. PMID- 15785932 TI - The distribution of microvascular response in Achilles tendonopathy assessed by colour and power Doppler. AB - OBJECTIVES: To assess the distribution of microvascular response on colour Doppler (CD) and power Doppler (PD) ultrasound (US) of the tendo Achilles (TA) in tendonopathy, and to look for any relationship between tendon morphology and symptoms. DESIGN AND PATIENTS: A retrospective, observational study was carried out on consecutive ambulant US patients with suspected tendonopathy, presenting with pain or an Achilles mass. Exclusion criteria were: use of steroids, and previous or possible rupture or surgery in either tendon or arthropathy. Using a 5-12 MHz linear array probe (ATL HDI 3000) both TAs were scanned. Tendonopathy was defined as tendon swelling and/or hypoechogenicity of the TA. The site, number and distribution of microvascularity, on CD and PD, and the anteroposterior size were recorded, with the analysis masked. RESULTS: Fifty-two patients presented with TA pain and six also with swelling. There were 34 males and 18 females, aged from 11 to 78 years (mean 45 years). Fifty-five TAs that showed tendonopathy with hypoechogenic areas were all observed to be over 5.9 mm (mean 11.1 mm, range 5.9-20 mm), of which 45 were symptomatic with abnormal PD and 24 with abnormal CD flow. It was observed that the extent and completeness of vessel branching was more extensive on PD than CD. All TAs demonstrating tendonopathy were over 5.9 mm in adults and all TAs that showed PD flow were over 6.5 mm. All microvessels originated towards the TA from the ventral surface usually into tendonopathy, and were 16-fold more frequent around the margins. There were 49 TAs with normal spectral US, and with no PD flow, with a mean size of 4.5 mm (range 3.0-7.4 mm). For the right and left TAs independently analysed and taking the 40 patients with a paired asymptomatic and symptomatic tendon: (1) There was a highly significant difference in size (P<0.00001) using the paired t test (parametric) between the asymptomatic tendon (mean 5.2+/-1.4 mm (1 SD)), and the contralateral morphologically abnormal and symptomatic side (mean 9.7+/-1.4 mm). (2) There was no linear Pearson correlation (0.25) between TA size and duration of symptoms (P=0.11) for symptomatic tendons. (3) There was a positive Spearman correlation (0.84) between the number of vessels and TA size (P<0.00001). (4) There was a significant difference in the number of PD vessels using the non-parametric Wilcoxon signed test (P<0.00001) between the symptomatic and asymptomatic groups. CONCLUSIONS: (1) PD shows more tendon microvascularity than CD in TA tendonopathy. (2) All microvessels arise on the ventral side of the TA. (3) There is a non-linear relationship between tendonopathy, TA size and the amount of microvascularity, but not between PD and duration of symptoms. (4) Morphologically abnormal adult TAs were larger than 5.9 mm, and PD flow was only seen in TAs above 6.5 mm. PMID- 15785934 TI - Increased activity of lysosomal enzymes in the peritoneal fluid of patients with gynecologic cancers and pelvic inflammatory disease. AB - PURPOSE: To investigate whether the activity of lysosomal enzymes is increased in the peritoneal fluid of patients with gynecologic cancers compared to activity in the peritoneal fluid from normal subjects and those with pelvic inflammatory disease, and fluid from benign ovarian cysts. PATIENTS AND METHODS: beta glucuronidase, beta-galactosidase, and alpha-mannosidase activity was measured in the peritoneal fluid from patients with gynecologic cancer, pelvic inflammatory disease, and normal subjects, and fluid from benign ovarian cysts. RESULTS: The mean+/-SD of beta-glucuronidase, beta-galactosidase, and alpha-mannosidase activity in the gynecologic cancers was 120+/-50 nmol, 203+/-86 nmol, and 240+/ 119 nmol 4-methylumbelliferone/ml/h, respectively; in the normal control subjects it was 22+/-9 nmol, 46+/-10 nmol, and 80+/-23 nmol, respectively (P=0.00003, 0.0001, and 0.0001, respectively). The activity was increased even in cases without malignant cells in the peritoneal fluid. In pelvic inflammatory disease it was 148+/-82 nmol, 278+/-112 nmol, and 291+/-140 nmol, respectively. The activity in the fluid of the ovarian cysts was similar to that of the normal peritoneal fluid. There was a significant positive correlation between enzyme activity and stage of cancer, that was stronger for beta-glucuronidase (r=0.889, P=0.003). CONCLUSION: The increased lysosomal enzyme activity in gynecologic cancers, without overlapping between patients and normal subjects or benign ovarian cyst fluid, indicates that such measurements might be applied for diagnostic purposes. PMID- 15785933 TI - The hypermethylation and protein expression of p16 INK4A and DNA repair gene O6 methylguanine-DNA methyltransferase in various uterine cervical lesions. AB - PURPOSE: This study is aimed at investigating the significance of gene promoter methylation status and protein expression of p16 INK4A and O6-methylguanine-DNA methyltransferase (MGMT) in the various uterine cervical lesions. MATERIALS AND METHODS: Methylation status by using methylation-specific polymerase chain reaction (MS-PCR) and protein expression by using immunohistochemistry for p16 INK4A and MGMT genes were performed in cervical squamous intraepithelial neoplasms (CIN), invasive squamous cell carcinomas (SCC), adenocarcinomas and non neoplastic cervices. RESULTS: None of 20 non-neoplastic cervices showed p16 INK4A and MGMT gene hypermethylation, whereas at least one of these genes was hypermethylated with 50.0% (5/10) of CIN I, 65.0% (13/20) of CIN II-III, 70.2% (33/47) of SCC and 85.0% (17/20) of adenocarcinoma. p16 INK4A protein was totally negative in non-neoplastic cervices, but positive with 90.0% of CIN I, 100% of CIN II-III and adenocarcinoma, and 78.7% of SCC. MGMT protein was expressed in 10% of non-neoplastic cervices, but significantly increased in SCC (42.5%) and adenocarcinoma (70.0%). The protein expression of p16 INK4A and MGMT was not related to their gene promoter methylation status. CONCLUSIONS: The hypermethylation of p16 INK4A and MGMT genes in the uterine cervix may indicate the presence of malignant cells, and p16 INK4A immunostaining is useful in grading CIN and diagnosing invasive SCC and adenocarcinoma. PMID- 15785935 TI - Dedifferentiated chondrosarcoma--a fatal disease. AB - BACKGROUND: We report on 13 patients with dedifferentiated chondrosarcomas. The mean age of the patients at diagnosis was 59.8 years. Nine patients were classified as stage IIB and four as stage III. METHODS: In 11/13 cases surgery was performed. Mostly, limb salvage with tumour resection and implantation of a megaprosthesis was done; three patients needed amputation or disarticulation. In one out of three patients with a pelvic tumour resection was followed by implantation of a pelvic replacement; the other two patients received tumour resection with autologous stabilisation of the pelvis. Surgical margins were wide in six patients, marginal in two and intralesional in three. Adjuvant chemotherapy was given to five patients. RESULTS: Recurrence was detected in 5/11 of the patients operated on: in two with wide, in one with marginal, and in two with intralesional resection. No recurrence was seen in 5/11 patients: in four after wide and in one after marginal resection. In one patient the stage was unknown. At follow-up 11 patients were dead of disease (DOD), one dead of unknown reason (DOU) and one alive with disease (AWD). The mean survival time was 9.7 months. Metastasis to different anatomical sites was evident after a period of 10 months. CONCLUSIONS: Our results resemble those reported in the literature. DDCS is rare and is the primary malignant bone tumour with the worst prognosis. Surgery is the most important procedure, although it is unclear whether a radical resection improves the long-term results. Information regarding neoadjuvant and/or adjuvant therapy with chemotherapy is very limited. PMID- 15785936 TI - The impact of ibuprofen on renal clearance in preterm infants is independent of the gestational age. AB - The aim of this investigation was to evaluate whether the relative change in renal clearance due to ibuprofen administration depends on the gestational age (GA). Clearance of amikacin was used as a surrogate marker for renal clearance in preterm infants (GA 24-34 weeks, day 1) on respiratory support. Clearance in infants co-treated with ibuprofen was compared to that of infants not co-treated with ibuprofen. Absolute change of clearance in four consecutive cohorts of increasing (<27, 27-28, 29-31, 32-34 weeks) GA and linear correlations of clearance on GA were calculated in both groups. Data were collected from 204 infants, of whom 93 were co-treated with ibuprofen. Mean amikacin clearance was lower (0.48 to 0.59 ml/kg/min, P <0.001) in infants co-treated with ibuprofen. Absolute decrease in mean amikacin clearance (0.10 to 0.14 ml/kg/min) was similar in all four cohorts. Linear correlations of clearance with GA were documented in infants co-treated (y =0.21+0.011x) or not co-treated (y =0.44+0.009x) with ibuprofen. Renal clearance, reflected by amikacin clearance, is decreased by ibuprofen in preterm infants of 24 to 34 weeks GA on respiratory support on the 1st day of life, independent of the GA. PMID- 15785937 TI - Hemihypertrophy, renal dysplasia and benign nephromegaly. AB - Hemihypertrophy is associated with malignant visceral abdominal neoplasms in childhood. Benign nephromegaly and nephroblastomas are both known to occur with hemihypertrophy; however, association with renal dysplasia has not previously reported. We describe an infant presenting with recurrent haematuria who had segmental hemihypertrophy, ipsilateral renal dysplasia and contralateral benign nephromegaly. Although debated, renal dysplasia may predispose to and predate malignant change. Rational management and optimal surveillance of renal dysplasia and benign nephromegaly in hemihypertrophy is discussed. PMID- 15785938 TI - Secondary renal Fanconi syndrome caused by valproate therapy. AB - Although renal Fanconi syndrome resulting from valproate (VPA) has occasionally been reported, the detailed clinical characteristics of this disease remain unclear. To clarify the clinical features of patients with VPA-induced Fanconi syndrome, we analyzed the clinical and laboratory data of seven affected patients. All patients were children, were severely disabled and required tube feeding. Five patients required treatment with multiple anticonvulsant agents. Hypophosphatemia and hypouricemia were found in all patients. Mild proteinuria, increased excretion of urinary beta2-microglobulin (beta2MG) and generalized hyperaminoaciduria were present in all patients. The renal biopsy of one patient exhibited tubulointerstitial nephritis without any structural abnormalities of the mitochondria in proximal renal tubular cells. All patients recovered from the Fanconi syndrome after the cessation of VPA therapy without any long-term renal sequellae. These results indicate that young age and being severely disabled with tube feeding and anticonvulsant polytherapy are contributory factors to the development of VPA-induced Fanconi syndrome. Serum phosphate and uric acid concentrations and urinary beta2MG levels in addition to serum electrolytes and urinalysis should be examined regularly in patients receiving VPA therapy, especially in those with the contributory factors outlined above. Patients with Fanconi syndrome caused by VPA have a favorable renal outcome. PMID- 15785939 TI - Renal involvement in Down syndrome. AB - Kidney disease has not been considered a frequent complication in Down syndrome (DS) patients; a variety of urological abnormalities and glomerulopathies have been reported in this population, and some DS patients develop chronic renal failure (CRF). The aim of this study was to improve the understanding of renal disease in patients with DS, focusing on the incidence and range of kidney and urological abnormalities in a population of DS patients. A cross-sectional study was carried out in DS patients referred from a pediatric genetics unit of a tertiary care center. Medical records were reviewed. A 24-h urine specimen and a blood sample were obtained. Fractional excretion of sodium and potassium, tubular reabsorption of phosphate, urinary excretion of calcium, magnesium, uric acid, creatinine clearance and proteinuria were determined. Ultrasound was performed to evaluate the kidneys and the urinary tract. Laboratory data were reviewed for any possible renal disorder. Sixty-nine patients, aged 12 months to 24 years, were recruited. Pathological findings included three cases of voiding disturbances and a case of hypertension in a 7-year old girl. Eight patients (11.6%) had hyperuricemia without gout. Eighteen patients (24.2%) had hyperuricosuria. Urinalysis revealed three cases of mild proteinuria and two patients with microscopic hematuria. Minor radiological abnormalities were found in five patients (7.3%). Three patients (4.5%) had CRF. Renal disease in patients with DS is not as rare as previously thought, although the majority of findings are of minor relevance. According to the variety of pathologies, and in order to detect early irreversible renal injury, it seems quite reasonable to perform regular monitoring of renal function in these patients. PMID- 15785940 TI - Megacalycosis: report of two cases. AB - Congenital megacalycosis is a rare renal disorder consisting of caliceal dilatation without evidence of renal pelvic or ureteral obstruction. Since its first description in 1963, only a few cases have been reported. The defect is mostly unilateral, shows male predominance and does not disrupt the renal functions. We report on two male patients with bilateral megacalycosis and different clinical presentations. The first case was diagnosed in adolescence during examination for a urinary tract infection and was followed-up uneventfully for nearly ten years. The second case was found to have megacalycosis during the assessment of antenatal hydronephrosis. We aimed to emphasize the importance of considering megacalycosis in the differential diagnosis of both antenatal hydronephrosis and other causes of caliceal dilatation to prevent unnecessary diagnostic and therapeutic interventions. PMID- 15785942 TI - Efficacy and safety of prolonged amlodipine treatment in hypertensive children. AB - To examine the long-term efficacy and safety of amlodipine in hypertensive children, data on prolonged use (> or = 6 months) of amlodipine in 33 children were reviewed. All children received amlodipine as sole therapy for their hypertension. Causes of hypertension included solid organ transplant (n=19), renal disease (n=7) primary hypertension (n=6), and drug-induced hypertension (n=1). Mean patient age at the start of amlodipine treatment was 9.8+/-4.8 years (range 1.3-16.9); there were 19 boys and 14 girls. Mean duration of amlodipine treatment was 20.4+/-11.5 months (range 6-48). Analysis of blood pressure and dosing data revealed that blood pressure reduction was sustained throughout the period of amlodipine treatment, while amlodipine dose remained stable (mean effective daily dose 0.17+/-0.12 mg/kg). No patient required discontinuation of amlodipine because of adverse effects. This small study suggests that prolonged amlodipine treatment is well tolerated in hypertensive children and provides sustained blood pressure control. Further studies are necessary to determine what effects if any long-term calcium channel blocker treatment has on the growth and development of children with hypertension. PMID- 15785941 TI - Intravenous calcitriol for treatment of hyperparathyroidism in children on hemodialysis. AB - This double-blind, placebo-controlled study evaluated the safety and efficacy of intravenous (i.v.) calcitriol (Calcijex) for treatment of secondary hyperparathyroidism (secondary HPT) in pediatric end-stage renal disease (ESRD) patients on hemodialysis (HD). After a 2 to 6-week washout period of all vitamin D compounds, patients with two consecutive PTH values > 400 pg mL(-1), calcium levels < or = 10.5 mg dL(-1) and calcium x phosphorus product values < or = 70 mg2 dL(-2) were eligible for the treatment phase. Patients received a bolus injection of calcitriol or placebo three times a week, immediately after dialysis for up to 12 weeks. Initial doses (0.5-1.5 microg) were based on the severity of secondary HPT. The dose was increased every two weeks by 0.25 microg until there was at least a 30% decrease in PTH from baseline, or Ca > 11.0 mg dL(-1), or Ca x P > 75 mg2 dL(-2). Overall, 11/21 (52%) patients in the calcitriol group had two consecutive > or = 30% decreases from baseline in serum PTH compared with 5/26 (19%) patients in the placebo group (P=0.03). The mean total alkaline phosphatase decreased from 274 to 232 IU L(-1) in the calcitriol group and increased from 547 to 669 IU L(-1) in the placebo group (P=0.002). The mean bone-specific alkaline phosphatase decreased from 72.5 to 68 microg L(-1) in the calcitriol group and increased from 105.3 to 148.5 microg L(-1) in the placebo group (P=0.03). The incidence of two consecutive occurrences of elevated calcium x phosphorus (Ca x P > 75 mg2 dL(-2)) product was higher in the calcitriol group than in the placebo group (P=0.01). Two consecutive occurrences of phosphorus > 6.5 mg dL(-1) occurred in 71% of the calcitriol group and 46% of the placebo group (P=0.14). Calcium levels > 10.5 mg dL(-1) were more common in the calcitriol group than in the placebo group (P=0.01). There was a direct relationship between serum phosphorus concentration and the percentage change in PTH from baseline in both the calcitriol group (r=0.46; P<0.0001) and the placebo group (r=0.21; P=0.0005). This study demonstrates that i.v. calcitriol, at initial doses of 0.5-1.5 microg, effectively reduces PTH levels in pediatric HD patients and that patients should be closely monitored for hyperphosphatemia and elevated Ca x P product. PMID- 15785943 TI - Artefacts in magnetic resonance imaging caused by dental material. AB - A common problem in computer tomography (CT) based imaging of the oral cavity is artefacts caused by dental restorations. The aim of this study was to investigate whether magnetic resonance imaging (MRI) of the oral cavity would be less affected than CT by artefacts caused by typical dental restorative alloys. In order to assess the extent of artefact generation, corresponding MRI scans of the same anatomic region with and without dental metal restorations were matched using a stereotactic frame. MRI imaging of the oral and maxillofacial region could be performed without reduction of the image quality by metallic dental restorations made from titanium, gold or amalgam. Dental restorations made from titanium, gold or amalgam did not reduce the image quality of the MRI sequence used in imaging of the oral and maxillofacial region for dental implant planning. In this respect MRI is superior to CT in implant planning. PMID- 15785945 TI - [Technique of arthroscopic stabilisation]. PMID- 15785944 TI - A standardised method for measuring magnetisation transfer ratio on MR imagers from different manufacturers--the EuroMT sequence. AB - Magnetisation transfer ratio (MTR) is increasingly used to evaluate neurological disorders, especially those involving demyelination. It shows promise as a surrogate marker of disease progression in treatment trials in multiple sclerosis (MS) but the value measured is highly dependent on pulse sequence parameters, making it hard to include the technique in large multi-centre clinical trials. The variations can be reduced by a normalisation procedure based on the flip angle and timing of the presaturation pulse, but correction for parameters such as saturation pulse shape, amplitude, duration and offset frequency remains problematic. We have defined a standard pulse sequence, to include a standard presaturation pulse and set of parameters, which can be implemented on scanners from both General Electric and Siemens, and has also been used on Phillips scanners. To validate the sequence and parameters, six European centres measured MTR in the frontal white matter of normal volunteers. It was possible to measure MTR values in controls which were consistent to within approximately +/-2.5 percentage units across sites. This degree of precision may be adequate in many situations. The remaining differences between sites and manufacturers are probably caused by B1 errors. PMID- 15785946 TI - [Anterior femoroacetabular impingement after healed femoral neck fractures]. AB - OBJECTIVES: To describe anterior femoroacetabular Impingement (AFAI) as a cause of persistent painful loss of motion and progressive joint-destruction in patients with a healed femoral neck fracture, and to evaluate results after its surgical treatment. METHODS: Eleven patients with groin pain elicited by motion and exertion following a healed femoral neck fracture were diagnosed clinically, by conventional radiographs and radial Arthro MRI with AFAI. During surgical subluxation or dislocation of the hip joint the impingement was visually verified and eliminated by re-shaping the anterior contour of the head-neck-junction. RESULTS: All patients presented a flat contour of the anterior head-neck-junction causing a cam-type impingement with subsequent damage of the anterior-superior acetabular cartilage adjacent to the rim. These chondral changes result from the repetitive compression and shear forces between the flattened head-neck junction and the acetabular cartilage in flexion and internal rotation. At five year follow-up a clear improvement of the symptoms was observed without any signs of progressive joint destruction. CONCLUSION: When chronic pain after a healed femoral neck fracture without necrosis of the femoral head occurs, the possibility of an AFAI caused by retrotorsion of the proximal fragment should be taken into consideration. The symptoms of AFAI can be relieved by surgical correction of the femoral head-neck-offset. The existing damage of the acetabular cartilage originated by the impingement at the time of surgery can hardly be improved. Therefore anatomical fracture reduction should be performed in order to prevent the development of osteoarthritis. Pre-existing deformities of the joint should be treated at the time of operative fracture treatment. PMID- 15785947 TI - Differential respirable dust related lung function effects between current and former South African coal miners. AB - Dust-related dose-response decrements in lung function among coal miners have been reported in several studies, with varying magnitudes across populations. Few studies have compared differences between current and former coal miners. No studies on dose response relationships with lung function have been conducted in South African coal mines, one of the top three producers of coal internationally. The objectives of this study were (1) to describe the relationship between respirable dust exposure and lung function among current and former South African coal miners and to determine whether differential dust related effects were present between these employment categories; (2) to examine dust related dose response relationships, controlling for potential confounding by smoking and a history of tuberculosis (TB). Six hundred and eighty-four current and 188 ex miners from three bituminous coal mines in Mpumalanga Province were studied. Interviews assessing work histories, smoking profiles and other risk factors were conducted. Work histories were also obtained from company records. Standardised spirometry was performed by trained technicians. Cumulative respirable dust exposure (CDE) estimates were constructed from company-collected sampling and measurements conducted by the researchers. Regression models examined the associations of CDE with per cent predicted FEV(1) and FVC, controlling for smoking, past history of TB and employment status. A statistically significant decline in FEV(1) of 1.1 and 2.2 ml/mg-year/m(3) was found in representative 40 year-old, 1.7-m tall current and former miners, respectively. Significant differences were found between the highest and medium exposure categories. Ex miners had a lower mean per cent predicted lung function than current miners for each cumulative exposure category, suggesting a "healthy worker" effect. Past history of TB contributed to 21 and 14% declines in per cent predicted FEV(1) and FVC, respectively. Thus, in this cohort, a dose-related decline in lung function was associated with respirable dust exposure, with a magnitude of effect similar to that seen in other studies and important differences between current and former employees. A "healthy worker" effect may have attenuated the magnitude of this relationship. TB was a significant contributor to lung function loss. PMID- 15785948 TI - Characteristics of wet work in nurses. AB - BACKGROUND OBJECTIVES: Nursing is known for its high prevalence of hand dermatitis, mainly caused by the intense exposure to wet work in nursing activities. We aimed to study the characteristics of wet work exposure in nursing. METHOD: Trained observers monitored the duration and frequency of different wet work activities in 45 randomly chosen nurses from different wards during a morning shift, using a method of continuous observation based on labour observation techniques. RESULTS: Wet work in intensive care units accounted for 24% of the overall morning shift duration, with a frequency of 49 incidents. This was 16% in dialysis wards, with a frequency of 30 incidents, and 9% on regular wards, with a frequency of 39 incidents. The wet work activities had short mean duration cycles. The mean duration of occlusion by gloves was 3.1 min on regular wards and 6.7 min in intensive care units. DISCUSSION: The characteristics of wet work in nurses differed substantially, depending on the ward. According to the German regulation TRGS 531, our observations classify nursing as a wet work occupation, due to the frequency of wet work rather than its duration. The mean duration of occlusion in our observations was short, which makes an occlusion induced irritating effect doubtful. Reduction in wet work exposure in nursing on regular wards could focus on the reduction of the frequency of hand-washing and patient-washing. We suggest increasing the use of gloves for patient washing. Although this will increase exposure to occlusion from gloves, it may reduce the frequency of exposure to water and soap by about a quarter. PMID- 15785950 TI - [Standard operating procedures--anaesthesiology, intensive medicine, pain therapy and emergency medicine exchange]. PMID- 15785949 TI - Treatment of myelodysplastic syndrome with isolated del(5q) including bands q31 q33 with a combination of all-trans-retinoic acid and tocopherol-alpha: a phase II study. AB - All-trans-retinoic acid (ATRA) alone or in combination with cytokines and vitamins has been shown to stimulate erythropoiesis in low-risk myelodysplastic syndromes (MDS). We performed a phase II study on 29 patients with MDS and isolated del(5q) including bands q31-q33 to determine the efficacy and safety of ATRA in combination with tocopherol-alpha. All patients had low/intermediate-1 risk MDS according to the international prognostic scoring system. They received 45 mg/m(2) ATRA on days 1 to 90, and 90 mg/m(2) on days 91 to 180. Tocopherol dosage was 600 IU three times daily. Twenty-four patients completed dose level I, and 12 patients dose level II. Eighty-six percent of patients experienced side effects. Thirty discontinued the drug treatment due to such events as skin reactions, cheilitis, conjunctivitis, joint pain, creatinine increase, or CNS symptoms. One patient (3%) achieved a major erythroid response resulting in transfusion independence throughout the study. Four patients (14%) achieved a minor erythroid response with >50% reduction of transfusion needs. None of the participants had a cytogenetic response. There was no significant improvement in quality of life among responding patients as measured by the European Organization for the Research and Treatment of Cancer (EORTC) quality of life questionnaire. Based on these results, the combination of ATRA and tocopherol alpha is not recommended for the treatment of del(5q) MDS. PMID- 15785951 TI - Interruption from irrelevant auditory and visual onsets even when attention is in a focused state. AB - The common view on the interplay between exogenous and endogenous orienting holds that abrupt onsets are not capable of attracting attention when they occur outside the current focus of attention. Does this also apply to sudden irrelevant auditory onsets and when irrelevant visual onsets occur far in the periphery? In addition, does focused attention also reduce the alerting effect of auditory onsets, or vice versa, do highly alerting stimuli distort the attentional state? Crossmodal and unimodal variants of the Posner paradigm were examined in two experiments with targets and irrelevant onsets occurring at 28.3 and 19.3 degrees from fixation. Either centrally presented arrows indicated the forthcoming position of visual targets to be discriminated, or warning cues signaled the likely moment of target occurrence. The targets could be preceded by peripheral auditory or visual onsets that were to be ignored. Crossmodal and unimodal exogenous orienting effects of these irrelevant onsets were observed while participants focused at the relevant side. In addition, no evidence was found that the alerting effect of auditory onsets was dependent on focused attention. Our findings indicate that, at least under the current conditions, neither crossmodal nor unimodal orienting effects of peripheral events dissipate when attention is in a focused state. PMID- 15785952 TI - Propagation velocity and triggering threshold of retinal spreading depression are not correlated. AB - Spreading depression (SD) is a pronounced but transient disturbance of cellular homeostasis in the neuropil of the central nervous system which spreads in a wave like manner across the tissue. At the wavefront the cells depolarize and a distinct ion redistribution between intra- and extracellular space is observed. In the aftermath of SD the recovering tissue is refractory: during an early absolute refractory period no further SD can be triggered, during the subsequent relative refractory period SD waves spread at lower velocity than usual. In this paper we shall examine the influence of temperature on SD triggering and on SD propagation in the chicken retina (retinal spreading depression, rSD) and we shall examine rSD triggering and rSD propagation in the refractory period. It will be shown that cooling decreases the threshold of rSD triggering, i.e. it becomes easier to trigger rSD when the temperature is reduced. At the same time cooling slows rSD propagation. In contrast, during the relative refractory period triggering rSD is more difficult than usual while rSD propagation is also slowed. These results demonstrate that the propagation velocity of rSD is not correlated with the triggering threshold. In particular, the propagation velocity of rSD must not be used to predict the influence of experimental conditions on the triggering threshold. PMID- 15785953 TI - Modification of cornea-evoked reflex blinks in rats. AB - Although maintaining the tear film on the cornea is the most important role of blinking, information about the organization and modification of cornea-evoked blinks is sparse. This study characterizes cornea-evoked blinks and their modification in urethane-anesthetized rats. Cornea-evoked blinks typically begin 16.2 ms after an electrical stimulus to the cornea and last an average of 50.2 ms. In anesthetized rats, the blink only occurs ipsilateral to the stimulus. In response to cornea stimulation, the orbicularis oculi EMG activity typically exhibits two bursts that correlate with the arrival of A delta and C-fiber inputs to the spinal trigeminal complex. In the paired-stimulus paradigm, suppression of the blink evoked by the second cornea stimulus occurs for interstimulus intervals less than 300 ms and is exclusively unilateral. Stimulation of the contralateral cornea does not affect subsequent blinks evoked from stimulation of the ipsilateral cornea. To determine whether activation of cornea-related neurons in the border region between the spinal trigeminal caudalis subdivision and the C1 spinal cord (Vc/C1) inhibits the second blink in the paired-stimulus paradigm, we examine the suppression of cornea-evoked blinks caused by microstimulation in this region. This suppression of orbicularis oculi EMG activity begins 8.3 ms after Vc/C1 stimulation. Activation of this region, however, is unlike suppression in the paired-stimulus paradigm because Vc/C1 activation bilaterally inhibits cornea-evoked blinks. Thus, activation of Vc/C1 is a previously unidentified mechanism for modulating cornea-evoked blinks. PMID- 15785954 TI - Correlation between 99mTc-(V)-DMSA uptake and constitutive level of phosphorylated focal adhesion kinase in an in vitro model of cancer cell lines. AB - PURPOSE: Although a number of prognostic indicators have been developed, it is still difficult to predict the biological behaviour of all cancer types.( 99m)Tc (V)-DMSA (V DMSA) uptake and focal adhesion kinase (FAK) expression and activation level could be potential agents for this purpose. We hypothesised the existence of a correlation between V DMSA, whose uptake is linked to phosphate ions, essential compounds for tumour growth and cell proliferation, and the adhesion protein FAK, whose elevated expression and level of constitutive activation are implicated in cancer progression. The aim of this study was to assess the relationship between V DMSA incorporation rate and FAK expression and activation by phosphorylation on tyrosine 397 residue. METHODS: We determined V DMSA uptake in six different cancer cell lines and we measured FAK expression and activation by using Western Blotting analysis. Correlations with factors known to be associated with poor prognosis, such as invasive potential, resistance to chemotherapy and proliferation rate, were also investigated. RESULTS: The cell lines exhibited different V DMSA incorporation rates. In addition, these cells showed the same FAK expression, but various degrees of activation. A correlation was observed between V DMSA uptake and level of FAK phosphorylation and between V DMSA or constitutive FAK activation and proliferation rate. However, no correlation was shown between these parameters and the other factors tested, i.e. invasive potential and anticancer drug resistance. CONCLUSION: The results of this in vitro study clearly demonstrate that phosphorylation of FAK, proliferation rate and V DMSA uptake are closely related. Because proliferation and a high level of constitutive FAK activation are linked to cancer progression, it can be assumed that in vivo V DMSA uptake reflects tumour aggressiveness. PMID- 15785955 TI - Dosimetry of 60/61/62/64Cu-ATSM: a hypoxia imaging agent for PET. AB - PURPOSE: Cu-diacetyl-bis(N4-methylthiosemicarbazone (Cu-ATSM) is an effective marker for the delineation of hypoxic tissue. Dosimetry calculations by the established Medical Internal Radionuclide Dose (MIRD) approach were performed with both animal and patient data. METHODS: Human absorbed dose estimates extrapolated from rat data were based on the biodistribution of 61Cu-ATSM in adult rats. Eighteen tissues were harvested and time-activity curves generated. The measured residence times and the MIRD S-values for 60Cu-ATSM were used to estimate human absorbed doses. The biodistribution of the tracer was directly measured in five patients injected with approximately 480 MBq of 60Cu-ATSM and imaged by positron emission tomography (PET) with a whole-body protocol. The combined data from all patients were used to derive organ residence times, and organ doses were calculated by MIRD methodology for 60Cu-ATSM, 61Cu-ATSM, 62Cu ATSM, and 64Cu-ATSM. RESULTS: Human absorbed dose estimates extrapolated from rat biodistribution data indicated that the kidneys appeared to be the dose-limiting organ (0.083 mGy/MBq) with a whole-body dose of 0.009 mGy/MBq. Based on the human PET imaging data, the liver appeared as the dose-limiting organ, with an average radiation dose of 0.064 mGy/MBq. The whole-body dose was 0.009 mGy/MBq and the effective dose was 0.011 mSv/MBq. CONCLUSION: These relatively small absorbed doses to normal organs allow for the safe injection of 500-800 MBq of 60Cu-ATSM, which is sufficient for PET imaging in clinical trials. PMID- 15785957 TI - Safety and feasibility of thyroid lobectomy via a lateral 2.5-cm incision with a cohort comparison of the first 50 cases: evolution of a surgical approach. AB - BACKGROUND AND AIMS: Over 500 minimal-access parathyroidectomies (MIPs) have been performed in our unit, and, from these, a technique for thyroid resection has evolved. We present a report on the evolution of minimal-access thyroid surgery (MATS) and compare the results with those from a cohort of patients operated on prior to the use of MATS. METHODS: We reviewed the evolution towards the MATS technique. The results of the MATS procedures were compared with those from an equal number of consecutive patients undergoing conventional lobectomy prior to the use of MATS. RESULTS: Fifty patients (mean age 45.6 years) underwent MATS between March 2002 and May 2004. The mean nodule diameter was 18.5 mm. In the MATS group there was one recurrent laryngeal nerve (RLN) injury, two temporary RLN neuropraxias, which recovered, and one haematoma. The control group (mean age 47.9 years) had a mean nodule size of 22 mm. In the controls there was one temporary RLN neuropraxia, which recovered, and two haematomas-P>0.05 (Fisher's exact test) for all the complications. CONCLUSIONS: MATS has evolved from an experimental approach into a safe and feasible surgical procedure based on the same operative approach as used for MIP. It provides an alternative to open thyroid surgery in appropriately selected cases. PMID- 15785956 TI - 18F-FDG PET in malignant lymphoma: significance of positive findings. AB - PURPOSE: The aim of this study was to evaluate the significance of increased uptake of 18F-fluorodeoxyglucose (FDG) in patients with malignant lymphoma (ML) studied by positron emission tomography (PET). METHODS: A total of 1,120 consecutive scans carried out in 848 patients were reviewed; all patients had a diagnosis of ML [574 non-Hodgkin's lymphoma (NHL) and 274 Hodgkin's disease (HD)] and were studied at completion of therapy, for suspected recurrence or during follow-up. PET was carried out after intravenous injection of 370 MBq of 18F-FDG; images were recorded after 60-90 min. Patients were selected whose reports indicated areas of increased FDG uptake. PET findings were considered positive for lymphomatous localisation when uptake occurred at sites of previous disease, in asymmetrical lymph nodes or in nodes unlikely to be affected by inflammation (mediastinal, except for hilar, and abdominal). PET findings were adjudged negative for neoplastic localisations in the following instances: physiological uptake (urinary, muscular, thymic or gastrointestinal in patients without MALT), symmetrical nodal uptake, uptake in lesions unrelated to lymphoma that had already been identified by other imaging methods at the time of PET scan, uptake at sites atypical for lymphoma, very low uptake and non-focal uptake. PET findings were compared with the results of other diagnostic procedures (including CT and ultrasound), biopsy findings and follow-up data. RESULTS: Overall, 354 scans (in 256 patients) showed increased FDG uptake (244 scans in NHL and 110 in HD): in 286 cases, FDG uptake was considered pathological and indicative of ML, in 41 cases the findings were described as uncertain or equivocal and in 37 cases, FDG uptake was considered unrelated to ML (in ten scans, concurrent findings of abnormal FDG uptake attributed to ML and uptake assigned to other causes were obtained) . Of the 286 patients with positive PET findings, 274 (95.8%) were found to have residual or recurrent ML (i.e. true positives). Four of the 41 patients with inconclusive findings turned out to have ML, while in 13 patients, pathological processes other than ML could be identified as the cause of FDG uptake. ML was excluded in all patients with findings reported as non pathological (100% true-negative rate). Therefore, the false-positive rate in our series was about 5%. The main cause of increased FDG uptake mimicking ML was inflammation. CONCLUSION: Our data confirm that 18F-FDG-PET has very high but not absolute specificity for ML. As already suggested, increased FDG uptake may also be observed in patients without active disease; in most cases, however, non pathological FDG accumulation is properly identified. Less frequently, inconclusive scans are encountered; these cases are usually caused by inflammation, which subsequently resolves. PMID- 15785958 TI - Haemodynamic responses to salbutamol and isometric exercise are altered in young adults with mild asthma. AB - OBJECTIVES: Tolerance to the haemodynamic effects of regularly inhaled beta(2) agonists has been reported in a research setting. It is unclear whether cardiovascular responses in adults with asthma are influenced by their use in routine clinical practice. This study aimed to characterise the effects of acute salbutamol administration on systemic haemodynamics in adults with mild asthma who were receiving intermittent beta(2) agonist treatment. METHODS: Ten patients with mild asthma and ten healthy age and sex-matched controls were recruited to a randomised placebo-controlled double-blind two-way crossover study. Each received salbutamol 200 mug or placebo, then performed sustained isometric handgrip exercise as an adrenergic cardiovascular stimulus. Heart rate and blood pressure responses were observed during rest and exercise, and cardiac index and systemic vascular resistance index were measured using non-invasive impedance cardiography. RESULTS: Isometric exercise caused a greater increase in heart rate (12+/-3 vs. 8+/-3 bpm, P<0.01) and diastolic blood pressure (19+/-3 vs. 12+/-3 mmHg, P<0.01) in patients with asthma compared with healthy controls. Salbutamol administration significantly increased heart rate, blood pressure and cardiac index in healthy controls, but these responses were attenuated in patients with asthma. CONCLUSION: These findings indicate that adults with asthma, who are receiving intermittent beta(2) agonist treatment, have a greater cardiovascular responses to sympathetic stimulation, and haemodynamic tolerance to acute salbutamol administration. PMID- 15785959 TI - CYP2C8 polymorphism frequencies among malaria patients in Zanzibar. AB - OBJECTIVE: The determination of the prevalence of the CYP2C8 main alleles in a typical set of malaria patients in Zanzibar, as these patients represent a typical population exposed to amodiaquine, an antimalarial mainly metabolized by CYP2C8. Also, to determine for the first time the frequencies of CYP2C8 alleles in native African populations. METHODS: Polymerase chain reaction-restriction fragment polymorphism for the identification of CYP2C8*1, CYP2C8*2, CYP2C8*3 and CYP2C8*4 on a random population of 165 unrelated malaria patients. RESULTS: The allele frequencies found were: CYP2C8*1 (wild type, 83.4%), CYP2C8*2 (13.9%), CYP2C8*3 (2.1%) and CYP2C8*4 (0.6%). In terms of genotypes, 70.4% of the patients showed the CYP2C8*1/ CYP2C8*1 genotypes, while heterozygous between the wild type and other minor alleles were seen in 26.0%. Finally, 3.6% of the patients were homozygous for slow metabolizer alleles. The frequencies observed are equivalent to those documented for African-Americans. CONCLUSIONS: CYP2C8 non-wild type alleles have a significant prevalence in the East African population studied. The consequent frequency of 3.6% of patients homozygous for slow metabolizer alleles represent a significant fraction of the population potentially in higher risk of adverse effects due to a less efficient metabolism of amodiaquine. As approximately 10(6) first-line treatments are currently performed in Zanzibar per year, this represents a non-negligible absolute number of amodiaquine exposures. This information constitutes a background for the pharmacovigilance programs presently being employed in Zanzibar. PMID- 15785960 TI - Effect of multiple-dose erythromycin on everolimus pharmacokinetics. AB - OBJECTIVE: We sought to quantify the influence of the CYP3A inhibitor erythromycin on the pharmacokinetics of everolimus, a CYP3A substrate. METHODS: This was a two-period, single-sequence, crossover study in 16 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 erythromycin 500 mg three times daily for a total of 9 days and a single 2-mg dose of everolimus coadministered on the fifth day of erythromycin therapy. The test/reference ratio and 90% confidence interval (CI) were derived for everolimus C (max) and AUC. RESULTS: During erythromycin coadministration, everolimus C (max) increased 2.0 fold (90% CI, 1.8-2.3) from 20+/-5 ng/ml to 40+/-10 ng/ml. Everolimus AUC increased 4.4-fold (90% CI, 3.5-5.4) from 116+/-37 ng h/ml to 524+/-225 ng h/ml. Everolimus half-life was prolonged by 39% from 32+/-6 h to 44+/-6 h. Erythromycin predose concentrations were not changed after single-dose administration of everolimus. CONCLUSION: Multiple-dose erythromycin increased single-dose everolimus blood levels by an average 4.4-fold (range, 2.0-12.6). During erythromycin treatment, a compensatory everolimus dose reduction should be made guided by everolimus therapeutic drug monitoring. PMID- 15785962 TI - A review of randomised clinical trials. PMID- 15785961 TI - Uptake of new drugs in rural and urban areas of Queensland, Australia: the example of COX-2 inhibitors. AB - OBJECTIVE: This study compared the changes over time in the volume of prescriptions of COX-2 selective inhibitors between rural and urban Queensland to reveal any difference in the uptake of the prescribing of these new drugs between two geographically distinct areas. METHODS: This study used data from an administrative claims database. Dispensing data were obtained for celecoxib and rofecoxib in two areas, one rural and one urban, defined by postcodes. The numbers of consumers in these areas were similar and they were served by similar numbers of general practitioners. The number of defined daily doses (DDDs) of celecoxib and rofecoxib dispensed at specific times was calculated. RESULTS: Statistical analysis revealed no significant difference between the total numbers of DDDs of COX-2-selective non-steroidal anti-inflammatory drugs dispensed in the rural and urban groups over the period August 2000 to December 2002 (P=0.81). The rate of uptake of usage was also clearly similar between the urban and the rural groups. Total usage peaked in August 2000 in both groups (urban 39 DDD/1,000 people per day; rural 37 DDD/1,000 people per day), coinciding with the pharmaceutical benefits scheme (subsidized) listing of celecoxib. The number of DDDs declined dramatically in the following month, and then peaked again in May 2002 (urban 34, rural 36). The number of DDDs then steadily decreased in both areas after October 2002. CONCLUSION: The results suggest that the marketing of the new COX-2 inhibitors and the patients' anticipation of a safe and effective treatment have overcome the geographical boundaries of Queensland. Both areas had very high rates of uptake of the prescribing of these new drugs. PMID- 15785964 TI - Reply to EJCP 2004-0275 and 2004-0350. PMID- 15785965 TI - Self-reported medicine use among adolescents from ethnic minority groups. PMID- 15785966 TI - Esophageal perforation complicating with spinal epidural abscess, iatrogenic or secondary to first thoracic spine fracture? AB - A 49-year-old drunken man was involved in a motorbike crash. He presented with cervical spine injury and multiple limbs fracture. Neuro-imaging demonstrated disruption of the C5-6 anterior longitudinal ligament, herniation of C3-4 and C5 6 discs, and Th1 compression fracture. The neurological deficits improved after anterior cervical decompression, fusion and fixation. One week later, he suffered from fever and severe upper back pain, and he developed paraplegia subsequently. The following image study disclosed esophageal perforation at the level of Th1 and epidural abscess spreading from levels Th1 to Th5. After proper management and rehabilitation, he achieved good recovery one year later at follow-up. We report the unique case of Th1 fracture with esophageal perforation complicated with spinal epidural abscess. The possible mechanism and the controversy concerning therapy for esophageal perforation are discussed. PMID- 15785969 TI - Analysis of Epstein-Barr virus gene expression upon phorbol ester and hydroxyurea treatment by real-time quantitative PCR. AB - Epstein-Barr virus (EBV) has the potential to undergo latent and lytic pathways during infection. However, expression of many of the viral genes during the lytic latent transition remains unclear. In this study, we investigated the effects of 12-O-tetradecanoylphorbol-13-acetate (TPA) and hydroxyurea (HU), two commonly used modulators of EBV life cycle, on the expression profiles of the entire genome of EBV persistent infected in B95-8 cells. After treatment with TPA for 48 h, the copy number of EBV genome in the cells increased about 2.5 fold, whereas HU treatment resulted in a reduction to approximately two-thirds of the original level. Except a small set of genes, the amounts of EBV mRNA are generally less abundant than that of beta-actin. The expression of a large fraction of the 80 EBV genes was found to be activated after TPA treatment with a noticeable increase of 19 and 21 fold, respectively in BSLF1 and BBLF4. In contrast, treatment of the B95-8 cells with HU, a nucleotide synthesis inhibitor, dramatically suppressed the expression of EBV lytic genes. In summary, we have demonstrated that real-time quantitative PCR is a reliable method to monitor the influence of drug-treatment in EBV genes regulation. Our results also provide a basis for further investigation on how the virus coordinates its own gene expression during latent-lytic pathway transition. PMID- 15785970 TI - Characterisation of Banana streak Mysore virus and evidence that its DNA is integrated in the B genome of cultivated Musa. AB - We have sequenced the complete genome of an isolate of Banana streak virus from banana cv. 'Mysore' and show that it is sufficiently different from a previously characterised isolate from cv. 'Obino l'Ewai' to warrant recognition as a distinct species, for which the name Banana streak Mysore virus (BSMysV) is proposed. The structure of the BSMysV genome was typical of badnaviruses in general, although ORF I had a non-conventional start codon. Evidence that at least part of the BSMysV genome is integrated in the B genome of cultivated Musa is presented and transmissibility by the mealybug Planococcus citri also demonstrated. PMID- 15785974 TI - [Early surgery in infantile esotropia]. AB - In Europe, early surgery for infantile esotropia is defined as surgery before 2 years of age for a strabismus of onset before 4 months. Subject to great controversy, the principal argument concerns binocularity, for some authors attainable with early surgery, for others an unreachable goal. In 2003, the European multicentric "Early versus Late Infantile Strabismus Surgery Study" compared the results of 2 groups of patients at 6 years of age, one having undergone early surgery, between the 6 (th) and the 24 (th) month of life (n = 231), the other later, between the 32 (nd) and the 60 (th) month of life (n = 301). Although slightly better in the early surgery group, the level of binocular functions in the vast majority of cases does not exceed that of subnormal or anomalous binocularity, while to achieve an equivalent postoperative angle the number of operations required was greater with early surgery. Aside from the above-mentioned study, cases of normal binocular single vision have long been reported after early surgery on both sides of the Atlantic. In North America it is sometimes believed that the earlier the surgery, the better the binocularity, some authors advising very early surgery, before 6 months of age. However, such reasoning is open to controversy since extremely early intervention increases the risk of including cases where the diagnosis is not fully established, in particular certain intermittent forms of strabismus that would explain some of the excellent binocular results. The virtue of being preventive rather than curative does not, however, offset the non-negligible fact that very early surgery does increase the risk of a greater number of operations. Indeed, very early surgery raises more questions than it answers, and the best age at surgery remains a subject open to debate. PMID- 15785975 TI - [Epiphora in cases of problems with eyelids and lacrimal ducts]. AB - This article illustrates the causes of epiphora and describes a systematic work up of the special eyelid and tear drainage disorders leading to this frequent symptom. PMID- 15785976 TI - [Optical coherence tomography in Malattia Leventinese]. AB - BACKGROUND: Malattia Leventinese (ML) is a genetically homogeneous macular dystrophy with an autosomal dominant mode of inheritance. Ophthalmoscopically it is recognisable by a radial pattern of drusen-like deposits in the macula and by parapapillary deposits, named Forni's verrucosities. The aim of this study is to describe optical coherence tomographic (OCT) findings and to compare them with histological data. PATIENTS AND METHODS: Six patients underwent ophthalmological examination, angiography and OCT. Diagnosis was confirmed by genetic analysis of the R345W mutation. A histopathological study of an ML donor eye was performed. RESULTS: OCT revealed a diffuse RPE-choriocapillaris thickening with nodular features in the macular and parapapillary areas. The protrusions reached as far as the outer nuclear layer. CONCLUSIONS: OCT is a non-invasive technique that provides a cross-sectional picture of the retina comparable to a histological section. In ML, OCT revealed a diffuse alteration of the RPE-Bruch's membrane complex. The macular and parapapillary nodular lesions are the tomographic equivalents of drusen and Forni's verrucosities. PMID- 15785977 TI - Syndromic retinitis pigmentosa: ERG and phenotypic changes. AB - PURPOSE: Our aim was to review the phenotype and extent of ERG changes in syndromic RP (SRP). PATIENTS AND METHODS: A retrospective review of charts of 82 patients seen over the last 20 years with SRP was carried out. Clinical data were compared with changes in ERG. Full-field ERGs comprised selective rod-driven, maximal dark-adapted mixed responses, and isolated cone-driven signals. Occasionally, ERGs were recorded under brief general anaesthesia. ERG changes were classified as normal, reduced or extinguished. RESULTS: Syndromic RP was diagnosed for the following entities: Usher (45 patients), Laurence-Moon-Bardet Biedl (LMBB, 17 patients), Kearns-Sayre (10), Batten (6), Refsum (3), Senior Loken (1). ERG changes varied in every subgroup and were therefore not specific for the syndromes. CONCLUSION: There was wide variation of clinical presentation in SRP, much as seen in isolated RP, often without obvious ophthalmoscopic changes. ERG testing is a prerequisite for differential diagnosis as well as for early detection of multiple handicaps. PMID- 15785978 TI - Asymmetry of the optokinetic nystagmus in lesion of the central nervous system. AB - BACKGROUND: Unilateral lesions of the central nervous system (CNS) may be associated with a reduction of the optokinetic nystagmus (OKN) slow component in the direction of the lesion. The aim of our study was to assess the role played in these cases by the direct injury of the OKN pathways and/or by a possible associated visual field defect. METHODS AND PATIENTS: Monocular OKN was elicited with black and white stripes moving temporally-to-nasally (TN) or nasally-to temporally (NT) at velocities of 15, 30, 45 and 60 degrees /s. Patients with cortical or chiasmal lesions associated with visual field defects were investigated. OKN was considered asymmetrical if the gain difference between TN and NT stimulation was not within 2 standard deviations of an age-matched control group (n = 86). RESULTS: We examined 12 patients with cortical lesions and 4 patients with chiasmal lesions. Asymmetric OKN gain was measured in 7 patients with cortical lesions associated with a visual field defect, and in 2 patients with chiasmal compression and bitemporal hemianopia. In 2 patients with isolated occipital lesions, OKN asymmetry was explained by the associated visual field defect. CONCLUSION: The interpretation of OKN asymmetry in patients with CNS lesions should not only consider a direct lesion of the OKN pathways but also a sensory deficit due to a visual field defect. PMID- 15785979 TI - Look and stare optokinetic nystagmus in healthy subjects and in patients with no measurable binocularity. A prospective study. AB - BACKGROUND: The aim of our study was to assess the difference between look and stare optokinetic nystagmus (OKN) in healthy subjects and in patients with early onset strabismus and no measurable binocularity. METHODS AND PATIENTS: OKN was elicited monocularly with black and white stripes moving horizontally or vertically at 15, 30, 45 and 60 degrees /s. Subjects were instructed either to follow individual stripes across the screen (look OKN) or to fixate stripes in the centre of the screen (stare OKN). We examined 20 healthy subjects (mean age 29 years; range 21 - 39), and 10 patients with no measurable binocularity (mean age 12.7 years; range 5 - 37). OKN gain was measured with photo-oculography. RESULTS: In both groups and at stimulation with the higher velocities gains for look OKN were significantly better than for stare OKN, and gains with horizontal stimuli were significantly better than with vertical stimuli (p < 0.05). In the group with no measurable binocularity horizontal look and stare OKN answers were significantly better for temporal-to-nasal stimulation than for nasal-to-temporal stimulation (p < 0.05). CONCLUSIONS: The performance of look or stare OKN influences the gain in healthy subjects and in patients with no measurable binocularity. Data about both modes of OKN stimulation are helpful to better interpret OKN responses especially in subjects with poor cooperation. PMID- 15785980 TI - Simultaneous fluorescein and indocyanine green angiography for exudative macular degeneration. AB - BACKGROUND: The present study investigates how often a simultaneous fluorescein and indocyanine green (ICG) angiography had therapeutic consequences and if it is useful as a clinical routine diagnostic tool. PATIENTS AND METHODS: 502 consecutive simultaneous angiographies in eyes with exudative macular degeneration were retrospectively studied. RESULTS: A classic extra- or juxtafoveolar choroidal neovascularisation (CNV) was found in 3.5 % of the eyes. A subfoveal predominantly classic CNV was present in 19 % of the angiographies. ICG angiography showed a vascular network in 3 % of the eyes with occult CNV in fluorescein angiography. A neovascularisation supplied by retinal vessels (retinal angiomatous proliferation) was found in 9 % and a polypoidal choroidal vasculopathy (PCV) in 6 %. Other plaques or hot spots were visible in 4 %. In 11 eyes with progressive exudation from PCV and threatening of the fovea, laser treatment was successfully performed. CONCLUSIONS: Combined angiography identifies treatable PCV. Advantages of a combined procedure (easier logistics, no missing of treatable cases) and arguments for a two step procedure with ICG angiography only in selected cases (lower costs, lower rate of adverse reactions) must be weighed against each other. PMID- 15785981 TI - [Cataract and combined cataract/glaucoma surgery as intraocular pressure-reducing procedure in eyes with relative anterior microphthalmos]. AB - PURPOSE: Eyes with relative anterior microphthalmos (RAM) (normal [> 20.0 mm] axial length but disproportionally small anterior segment [horizontal corneal diameter < 11 mm]) often present with increased intraocular pressure. Under these difficult conditions cataract surgery alone or in combination with a glaucoma filtering procedure is often required. PATIENTS AND METHODS: We retrospectively reviewed the records of 71 patients (108 eyes) with RAM in whom cataract only (n = 72) or combined cataract/glaucoma surgery (n = 36) was performed. Preoperative clinical and biometric findings, intraoperative complications and postoperative results (visual acuity and intraocular pressure) were analysed. RESULTS: Median preoperative values were 7.46 mm for corneal curvature, 2.53 mm for anterior chamber depth and 22.44 mm for axial length. The implanted lens had a median 25.0 dioptric power. The IOP was reduced from 15.6 +/- 3.9 mm Hg to 13.5 +/- 3.1 mm Hg with cataract surgery alone and from 22.4 +/- 5.5 mm Hg to 15.4 +/- 4.0 mm Hg with combined surgery. Mean best corrected visual acuity improved from 0.4 +/- 0.2 to 0.79 +/- 0.25 with cataract surgery and from 0.5 +/- 0.3 to 0.81 +/- 0.24 with combined surgery. Intraoperatively only 3 eyes had a rupture of the posterior capsule and postoperatively 2 eyes developed malignant glaucoma in the combined group. CONCLUSIONS: The results show that, in eyes with RAM, cataract surgery alone or in combination with glaucoma surgery can be performed with excellent results and low complication rate if adequate precautions are taken (especially the use of hyperosmotic agents). PMID- 15785982 TI - [Diagnosis of horner syndrome by pupil dilation lag]. AB - BACKGROUND: In the first seconds after switching off the lights, Horner pupils often show a dilation lag. This dilation lag is diagnostic for Horner syndrome. We investigated whether the visibility of the dilation lag can be improved by increasing the amplitude of pupil dilation. PATIENTS AND METHODS: In four patients with pharmacologically confirmed Horner syndrome pupil sizes were registered bilaterally simultaneously with an infrared camera. Dilation lag was induced by switching off the light while the patient was fixating at distance or by turning off the light and changing fixation from near to far at the same time. An increase of anisocoria of 0.5 mm or more within the first 4 seconds was interpreted as dilation lag. RESULTS: Dilation lags were seen in one patient with both methods, in two patients only with the second method, and in one patient with neither method. CONCLUSIONS: Dilation lag visibility can be improved by asking the patient to change fixation from near to far at the moment when the lights are switched off. PMID- 15785983 TI - [Endogenous Candida endophthalmitis combined with severe general diseases]. AB - BACKGROUND: Endogenous Candida endophthalmitis is a rare disease with increasing frequency and poor prognosis. PATIENTS AND METHODS: The course of endogenous Candida endophthalmitis in 7 eyes of 5 patients (age 2 months to 76 years) was evaluated. Underlying general diseases were diagnosed as colon cancer, diverticulitis, pancreatic insufficiency (with subclavian catheter), ileus and diabetes mellitus. Diagnosis was based on the very typical ocular feature combined with a positive blood or vitreous culture. Intensive antimycotic drug therapy was initiated and pars plana vitrectomy performed as soon as possible. RESULTS: The delay between onset of ocular symptoms and diagnosis amounted to one week and 2 months. In 3 eyes of 2 patients no vitrectomy could be done because of the very impaired state of health. These patients died of their general diseases one week and 2 months, respectively, later. During follow-up (4 weeks to 51 months) three eyes reached visual acuity of 5/10, 4/10 and 1/10. One eye reached 1/20 after additional surgery because of retinal detachment. In all vitrectomized eyes the diagnosis was substantiated by a positive culture of vitreous fluid. No recurrence of ocular inflammation was observed. CONCLUSIONS: Early vitrectomy seems to be mandatory in each case which is suspected of Candida endophthalmitis. Only with this option it is possible to fix the diagnosis and initiate adequate therapy in due time in order to improve the original poor prognosis. PMID- 15785984 TI - [Arnold-Chiari I malformation: three unusual manifestations]. AB - BACKGROUND: Arnold-Chiari Syndrome I is a malformation of the cervicomedullary junction, manifesting usually with downbeat nystagmus, palsy of the caudal cerebral nerves, headache, and vertigo. PATIENTS AND METHODS: We present three patients with unusual symptomatology. RESULTS: A two-year-old child with isolated non-ocular torticollis, a 52-year-old male patient, and a 42-year-old female patient, both with gaze-evoked nystagmus, underwent a cerebral MRI examination. The findings of the first two patients were typical for an Arnold-Chiari syndrome. The malformation in the third patient was found only by reviewing the initial MRI. CONCLUSIONS: Arnold-Chiari malformation may manifest atypically. An important step in the work-up of these patients is to ask the neuroradiologist to include the cervicomedullary junction in his evaluation. PMID- 15785985 TI - [Surgical management of monocular torsion and head tilt after macular translocation]. AB - BACKGROUND: Macular translocation following 360 degrees retinotomy is a possible surgical treatment of patients with age-related macular degeneration. However, it produces important subjective disturbances with diplopia and head tilt due to cyclodeviation. Complex surgical procedures involving both oblique muscles and two or four recti have been advocated. PATIENTS AND METHODS: Four symptomatic patients with macular translocation underwent counter-rotating surgery by very large recession and advancement of both oblique muscles of the affected eye. Preoperative subjective cyclo-deviations varied between 25 degrees and 60 degrees and head tilt ranged between 25 degrees and 45 degrees . RESULTS: Reduction of cyclodeviation ranged between 22 degrees ant 30 degrees in our four patients. Two patients showed residual cyclo-deviations of 3 degrees and 7 degrees with complete recovery of the head tilt. The remaining two patients showed significant subjective improvements and important reductions of head tilt, in spite of a large residual cyclodeviation. No patient exhibited binocular vision. CONCLUSION: Without jeopardizing the anterior segment blood supply of these elderly patients, a combined surgical procedure on both oblique muscles has shown to be sufficiently effective in reducing subjective cyclodeviation and head tilt in four cases of macular translocation. A complete counter-rotation does not seem to be required to achieve an important improvement of subjective symptoms. This may be due to sensorial adaptation. PMID- 15785986 TI - [Loa loa infection of the eye -- a case series]. AB - BACKGROUND: With increasing migration tropical diseases such as Loa loa infections are becoming more frequent in Europe. While the ocular diagnosis is usually straight forward, systemic work-up and treatment requires an interdisciplinary approach. We review the diagnostic and therapeutic work-up of ocular Loa loa infections based on a series of 4 cases that presented between 1998 and 2004. HISTORY AND SIGNS: The first symptoms in all cases were ocular irritations occurring 2 months to 8 years after a trip to West Africa. One case presented with a swollen upper eyelid without a visible worm. In three patients microfilariae were detected in the blood. THERAPY AND OUTCOME: In two cases visible subconjunctival worms could be removed under the slit lamp. Three cases required systemic treatment as inpatients while one case could be observed without systemic treatment. All 4 cases had a favourable outcome with complete eradication of the disease. CONCLUSION: Surgical removal of adult Loa loa worms from the subconjunctival space only improves the ocular symptoms. An interdisciplinary approach (ophthalmology, infectious disease and parasitology) for a systemic work-up and treatment is usually required. PMID- 15785987 TI - Pasteurella multocida endophthalmitis: case report and review of the literature. AB - BACKGROUND: Postoperative bacterial endophthalmitis is caused by the patient's endogenous flora in most cases (80 %). Pasteurella multocida (PM) is a Gram negative coccobacillus found in the upper respiratory tract of dogs and cats and is very rarely implicated in postoperative endophthalmitis. HISTORY AND SIGNS: We describe a case of PM endophthalmitis that developed after cataract surgery. THERAPY AND OUTCOME: Cultures of both the conjunctiva and the aqueous humor were positive for PM. Topical, intravitreous and intravenous antibiotics were administered. Despite treatment, the outcome was unfavourable and complicated by a corneal perforation. CONCLUSIONS: The prognosis of postoperative PM endophthalmitis remains poor, despite adequate treatment of the infection. A history of recent pet exposure should alert physicians to this possible aetiological factor. PMID- 15785988 TI - [Treatment of a complication after surgical removal of an aneurysmal cyst of the ethmoid]. AB - INTRODUCTION: An aneurysmal cyst (AC) is a rare benign bony tumor with a possible aggressive extension. We present a complication following the surgical ablation of an ethmoidal AC. CLINICAL HISTORY AND FINDINGS: A 40-year-old man with a left ethmoidal AC extending to the orbital roof underwent 2 surgeries. After the second one involving a neuro-surgical approach, a bilateral palsy of the superior oblique muscles (SO) appeared. The diplopia did not improve following a bilateral asymmetrical recession of the inferior recti muscles done elsewhere. There was an excyclotorsion up to 20 degrees in down gaze and a vertical deviation of 10 degrees in primary position. THERAPY AND OUTCOME: We performed a bilateral tucking of the anterior SO fibres with, on the left, an advancement of the inferior rectus and a resection of the medial rectus muscles. Two weeks after surgery the absence of cyclotorsional deviation allowed a binocular vision. CONCLUSION: The double vision due to the excyclotorsion, which was the main complaint, could be alleviated by an anterior strengthening of the SO. A precise measurement of the cyclotorsion is required for the surgical procedure. PMID- 15785989 TI - Can eyelashes migrate? AB - BACKGROUND: Intraocular cilia after penetrating injuries or surgery causing acute or subclinical inflammation are a well-known problem. In a healthy young patient with acute intraocular inflammation but no history of trauma the diagnosis may be missed initially. HISTORY AND SIGNS: A young farmer presented with severe eye pain, scleritis and a circumscribed chorioretinal and vitreous infiltrate. There was no history or evidence of eye trauma or systemic disease. THERAPY AND OUTCOME: Staphylococci were identified from vitreous material. During vitreous surgery an intraretinal cilium was found and removed. After intravitreal antibiotic treatment, the vision completely recovered. CONCLUSION: Acute intraocular inflammation should alert the ophthalmologist to consider an intraocular foreign body as a possible cause even if there is no history of trauma. PMID- 15785990 TI - Spasm of accommodation in a patient with increased intracranial pressure and pineal cyst. AB - BACKGROUND: In the setting of a normal neurological examination, accommodation spasm is frequently attributed to a non-organic etiology. Occasionally, organic disorders are associated. In particular, central lesions involving the dorsal midbrain and quadrigeminal plate have been described with disorders of accommodation. HISTORY AND SIGNS: A 36-year-old woman with idiopathic intracranial hypertension (IIH) had visual blur from pseudo-myopia due to accommodative spasm. Magnetic resonance imaging (MRI) revealed a pineal cyst that was reported to be an incidental finding. The patient had persistent papilledema and recurrent episodes of unilateral and bilateral visual blur from accommodative spasm despite medical management. THERAPY AND OUTCOME: A lumboperitoneal shunt effectively lowered her intracranial pressure (ICP). Thereafter, all symptoms of increased ICP, accommodative spasm and papilledema resolved. A functional, non organic cause for accommodation spasm was not suspected. CONCLUSIONS: To our knowledge, this is the first report of isolated accommodative spasm as a presenting symptom in a patient with IIH. The patient's accommodative spasm resolved with lowering of the ICP. It remains speculative whether her pineal cyst played a role in triggering the accommodative spasm. PMID- 15785991 TI - [Toxic optic neuropathy due to cisplatin therapy: a case report]. AB - BACKGROUND: Optic neuropathies presenting as bilateral disc oedema lead to a vast field of differential diagnosis; a toxic aetiology should be taken into consideration. HISTORY AND SIGNS: A 55-year-old patient with a non-metastatic pleuramesothelioma received cisplatin as palliative chemotherapy. Due to a unilateral high myopia on the left eye the patient has a severe anisometropic amblyopia. The patient complained about visual loss and changed colour vision. Visual acuity was 0.1 on the right eye and counting fingers on the left eye. The anterior parts of the eyes were unremarkable, the optic nerve head showed bilateral disc oedema. The MRI was free of tumour or signs of elevated intracranial pressure. Colour vision test Panel D-15 revealed a pathologic colour perception affecting the tetartan-axis. Perimetry showed a cecocentral and superior bundle scotoma. THERAPY AND OUTCOME: The patient has been treated with methylprednisolone (Solu-Medrol) 500 mg i. v. 1 gr daily for three days, continued by 100 mg oral steroids successively reduced over a period of three weeks. After three weeks visual acuity increased to 0.3. CONCLUSIONS: Toxic neuropathies including disc oedema, retinal oedema and optic neuritis are rare, but have been described as occasional side effects of treatment with cisplatin. PMID- 15785992 TI - Optic atrophy as a sign of wolfram syndrome. AB - BACKGROUND: The hallmark of Wolfram syndrome -- an extremely rare autosomal recessive disorder -- is the association of juvenile onset diabetes mellitus with optic atrophy. Additional symptoms such as deafness and diabetes insipidus may develop. HISTORY AND SIGNS: An 18-year-old man suffered progressive visual loss over the last seven years. Severe optic atrophy with extensive cupping and slightly elevated intraocular pressure in both eyes were found. Perimetry revealed peripheral defects with paracentral scotomas. ERG results excluded tapetoretinal degeneration; visual evoked responses were delayed. At the age of nine the patient had been diagnosed with diabetes mellitus type I and neurosensory deafness, seven years later a beginning hypogonadism was found. THERAPY AND OUTCOME: Wolfram syndrome is a progressive degenerative disorder with reduced life expectancy. Treatment is symptomatic. CONCLUSIONS: Wolfram syndrome should be considered in every patient who presents with optic atrophy and juvenile onset diabetes mellitus. PMID- 15785993 TI - [Wooden spike orbital injury]. AB - A 71-year-old female patient fell in her garden, inducing a skin wound on the temporal left eyebrow. Skin disinfection and wound closure were performed elsewhere, an X-ray image did not reveal a foreign body. She was referred to our service three days later with a progressive left periorbital swelling. Clinical inspection demonstrated a painfully, fluctuant swelling around the wound with an inflammatory pseudoptosis of the left eye. Vision was reduced on the left eye; anterior and posterior segments of both eyes were unharmed. After opening the wound sutures a purulent liquid was drained and a wooden fragment was found, measuring 22 x 0.5 mm. Because of restriction of abduction of the left eye, magnetic resonance imaging (MRI) was performed, detecting another organic intraorbital foreign body and a fracture of the left medial orbital wall. Anterior orbitotomy was performed and a wooden fragment was removed, measuring 47 x 0.6 mm. Under administration of intravenous antibiotics vision and ocular motility recovered uneventfully. This case emphasizes the value of MRI in the diagnostics of retained wooden foreign bodies as well as the importance of a soigneuse inspection of skin wounds with a high risk for remaining foreign bodies. PMID- 15785994 TI - Lethal Epstein-Barr virus associated NK/T-cell lymphoma with primary manifestation in the conjunctiva. AB - BACKGROUND: Lethal midline granulomas (LMG) are very rare angiocentric NK/T-cell lymphomas in association with Epstein-Barr virus. LMG are reported mainly in East Asia occurring in immune compromised patients. HISTORY AND SIGNS: A 41-year old male patient presented with a conjunctival swelling of his upper left eyelid. The lesion had increased over a period of 2 months despite topical corticosteroid treatment. Conjunctival biopsy revealed a highly malignant, CD3 + and BCL2 + extranodal T-cell lymphoma with features of an NK/T-cell origin (CD56 +, TIA + TCR-rearrangement: germline). All lymphoma cells were positive for Epstein-Barr virus RNA. The proliferation rate was highly elevated at 100 %. THERAPY AND OUTCOME: Systemic 1 (st) cycle chemotherapy with cyclophosphamide, doxorubicin, vincristin and prednisone resulted in a complete remission of the swelling within 4 days. However, one week later a massive conjunctival tumour reappeared with only partial regression after combined chemo- and radiotherapy. The patient died within a month because of untreatable pancytopenia due to malignant bone marrow infiltration. CONCLUSIONS: LMG is a rare but highly malignant Epstein-Barr virus associated NK/T-cell lymphoma that can occur in healthy, immune competent Caucasians. This is the first reported case of an LMG in an immune-competent Caucasian patient with primary ocular manifestation. The LMG has a high mortality rate despite systemic treatment and can be lethal within a few months or even weeks. PMID- 15785995 TI - Blackening of a choroidal hemangioma after photodynamic therapy. AB - BACKGROUND: Photodynamic therapy has recently advanced to a first line treatment of symptomatic circumscribed choroidal hemangiomas. HISTORY AND SIGNS: A sixty year-old male patient was referred with progressive visual loss in his left eye. Visual acuity measured 20/80 and fundus examination revealed a prominent red subretinal lesion superior to the fovea with adjacent pigment epithelial irregularities. Thickening of the central retina was seen on OCT imaging. Sonography revealed a dome-shaped lesion with a maximal thickness of 3.3 mm and high internal reflectivity. Combined fundus indocyanine green and fluorescein angiography showed early filling of large vessels within the tumor. THERAPY AND OUTCOME: The hemangioma was treated with verteporphin photodynamic therapy. Two days after treatment, with vision unchanged, fundus examination revealed blackening of the tumor. Two months later vision increased to 20/40 and the tumor area was atrophic and bright red with a slight hyperpigmented rim. Angiography revealed the disappearance of tumor vessels and choroidal hypoperfusion in the area of treatment. CONCLUSIONS: Choroidal hemangiomas may darken initially after photodynamic therapy. This is most probably caused by large coaguli within the voluminous lacuna-like blood-filled vessel system of the hemangioma that are induced by photothrombosis. PMID- 15785996 TI - Clinical and electrophysiological course of acute syphilitic posterior placoid chorioretinitis. AB - BACKGROUND: Syphilis can affect the anterior as well as the posterior segment of the eye at any stage. HISTORY AND SIGNS: A 52-year-old man was referred to our clinic because of acute loss of his vision and hearing. Best corrected visual acuity (BCVA) was counting fingers (CF) right and 0.05 left eye, respectively. Fundoscopy revealed bilateral intraretinal macular haemorrhages and a large yellowish edematous lesion involving macula and midperipheral retina. Fluorescein angiography showed diffuse late edema. The visual field showed a defect corresponding to the edematous lesion. The "Ganzfeld" electroretinogram (ERG) was markedly reduced and the multifocal ERG in the affected area was not recordable. THERAPY AND OUTCOME: VDRL, FTA-Abs and TPHA tests in serum and cerebrospinal fluid were positive. Intravenous penicillin therapy was initiated. BCVA, visual field and ERG eventually recovered completely. CONCLUSIONS: Syphilis should be considered as a differential diagnosis in any unclear inflammatory ocular condition. Awareness of syphilitic posterior placoid chorioretinitis allows rapid diagnosis and therapy. PMID- 15785997 TI - Varicella-zoster virus retinitis: successful evolution with a combination of antiviral therapies. AB - BACKGROUND: We present the description of a successful outcome in a case of varicella-zoster virus (VZV) acute retinal necrosis (ARN). HISTORY AND SIGNS: A healthy 40-year-old patient was admitted for a VZV retinitis. THERAPY AND OUTCOME: 10 days after the onset of intravenous (i. v.) acyclovir treatment, new small peripheral retinal necrotic lesions appeared in the right eye. A viral resistance was suspected and the acyclovir therapy was optimised with i. v. foscarnet combined with 2 intravitreal injections of ganciclovir. The outcome was favourable with a final vision of 1.0 after a follow-up of 30 months. No systemic or local complications were observed. CONCLUSIONS: VZV ARN is a severe infection with a poor prognosis. This case demonstrates that combination of antiviral therapies given intravenously (acyclovir + foscarnet) and in the vitreous (ganciclovir) may be safe and efficacious in the management of necrotising herpetic retinopathies affecting immunocompetent patients. PMID- 15785998 TI - [Pepper spray injuries of the anterior segment of the eye]. AB - BACKGROUND: A wide variety of pepper sprays is currently available and gaining increasing popularity among both professional guardians and amateurs. Adverse side effects to the anterior segment of the eye are known but underestimated. HISTORY AND SIGNS: We present two cases with severe corneal and conjunctival damage after accidental self injury by a pepper spray (Jet Protector Guardian Angel), benzyl alcohol 90.1 %, capsaicinoids 2.6 %). THERAPY AND OUTCOME: Despite immediate and intensive irrigation, a complete epithelial defect, extensive ischemia to the limbus and the conjunctiva and a circular conjunctival chemosis were diagnosed. After slow re-epithelialization in both cases, a neurotrophic superficial keratitis, a reduced corneal sensibility and in one case deep stromal scarring were noted. CONCLUSIONS: Pepper spray application to the eye might result in severe and permanent damage to the corneo-conjunctival tissue which is not adequately addressed in the current literature. From the present case reports arise the discussion whether the irritative and lipophilic capsacin/benzyl alcohol mixture or the pyrotechnical additives nitrocellulose und sinoxide are responsible for the anterior segment injuries. PMID- 15785999 TI - Idiopathic cyclitic retrolental membrane in children. AB - BACKGROUND: Cyclitic retrolental membranes (CRM) in children are usually associated with chronic uveitis or genetic syndromes. We report two rare cases of idiopathic CRM. PATIENTS AND METHODS: Two girls aged 9 and 13 years with visual acuities (VA) of 0.05 underwent lensectomy and anterior vitrectomy with dissection of the central part of the retrolental membrane and intraocular lens (IOL) implantation. RESULTS: The clinical evolution was excellent for the 9 year old girl who recovered 1.0 VA after 2 months. Histological examination revealed a fibroelastic tissue of unknown origin without inflammatory components. The 13 year old girl showed VA of 0.6 within 1 month. However, a recurrent CRM developed with retinal detachment and proliferative vitreoretinopathy (PVR). Vitrectomy, complete excision of the CRM and 360 degrees retinotomy with silicon oil tamponade attached the retina with limited visual recovery. Histology showed fibrovascular tissue with inflammatory components infiltrating the CRM. CONCLUSIONS: Idiopathic CRM in children are rare and can be composed of different histological tissues with very different clinical outcomes. PMID- 15786000 TI - Bilateral peripapillary subretinal neovessel membrane associated with chronic papilledema: report of two cases. AB - BACKGROUND: Increased intracranial pressure (ICP) usually results in headaches and papilledema with infrequent loss of visual acuity. Peripapillary subretinal neovessel membranes (PSNVM) rarely complicate chronic papilledema. HISTORY AND SIGNS: Case 1: A headache-free 9-year-old boy presented with a bilateral loss of vision due to chronic papilledema. Bilateral PSNVM was present. Cerebral imaging revealed a pilocytic astrocytoma and hydrocephaly. Case 2: A headache-free 39 year-old man presented with severe visual loss of the left eye due to an exudative macular star. Severe papilledema and PSNVM were present in both eyes due to idiopathic intracranial hypertension. THERAPY AND OUTCOME: Lowering of ICP (tumor resection - Case 1 and lumboperitoneal shunt - Case 2) resulted in involution of the PSNVMs in both patients. No improvement of visual acuity occurred for Case 1 (optic atrophy) but Case 2 benefited from gradual visual recovery. CONCLUSIONS: PSNVMs are rare complications of chronic papilledema, as only 10 patients have been reported so far in the literature. Absence of headaches in both of our patients was probably responsible for the late diagnosis, hence chronicity of papilledema. Laser therapy of PSNVM might not be necessary once the ICP is lowered. PMID- 15786001 TI - [Fundus quiz 2004]. AB - Six cases with characteristic fundus pathologies are presented and discussed using multiple choice questions. PMID- 15786003 TI - Improved spinal cord perfusion during thoracoabdominal aortic repair. AB - OBJECTIVES: The study's aim is to evaluate whether intraoperative neurophysiological monitoring with transcranial motor-evoked potentials (tcMEP) permits early recognition of neuronal lesions, thus making interventions to prevent irreversible spinal cord damage possible. MATERIAL AND METHODS: TcMEP monitoring was carried out in twelve patients (mean age 60 years) during open surgical thoracoabdominal aortic replacement. Current approaches for corrective, spinal cord-protecting interventions consist of: raising distal perfusion by increasing cardiopulmonary bypass (CPB) flow, catecholamine application, reducing central venous pressure, reimplantation of segmental arteries, and cerebrospinal fluid (CSF) drainage. RESULTS: Nine patients exhibited loss of tcMEP after segmental aorta clamping. In five patients we observed a recovery of tcMEP through counteractive measures. Three patients died intraoperatively, one patient presented with postoperative paraplegia and loss of tcMEP. CONCLUSION: TcMEP loss is associated with spinal cord ischaemia, causing postoperative paraplegia. TcMEP monitoring is an excellent method to detect spinal cord ischaemia at an early stage. PMID- 15786004 TI - Moderate hypothermia (30 degrees C) for surgery of acute type A aortic dissection. AB - OBJECTIVE: Antegrade cerebral perfusion has proved to be a reliable method of brain protection during surgery of thoracic aneurysms. In addition, the drawbacks of deep hypothermia may be avoided. This study examines the outcome after surgery for acute type A aortic dissections (AAD) using moderate (30 degrees C) systemic hypothermia compared with conventional techniques of cerebral protection. METHODS: Between January 1999 and August 2003, 74 patients underwent repair of acute type A aortic dissection. Moderate systemic hypothermia (30 degrees C) with selective antegrade cerebral perfusion through subclavian artery (group A) was used in 18 patients. Deep hypothermia (20 - 24 degrees C) was employed using either retrograde (18 patients, group B) or antegrade (38 patients, group C) cerebral perfusion. Tube graft replacement was performed in 55, valve-sparing procedure in 8, and composite graft replacement in 11 patients. RESULTS: The 30 day mortality was 5.5 % in group A, 5.5 % in group B, and 15.8 % in group C (A vs. C and B vs. C; p < 0.01). New postoperative permanent neurologic deficit occurred in 5.5 % of patients in group A, 16.7 % in group B, and 13.2 % in group C. Mean chest tube drainage within the first 24 h in groups A, B and C was 703 +/ 338, 1178 +/- 820, and 1447 +/- 802 ml, respectively (A vs. B and A vs. C; p < 0.01). Cardiopulmonary bypass, ICU, and hospital times were significantly shorter in group A. CONCLUSIONS: Selective antegrade cerebral perfusion with moderate systemic hypothermia appears to be a safe and sufficient tool for brain protection during AAD repair. In avoiding deep hypothermia, this technique may help to reduce cardiopulmonary bypass time and hypothermia-related side effects. PMID- 15786005 TI - Hypothermia-induced platelet aggregation: no effect of aprotinin (trasylol) but inhibition by eptifibatide (integrilin). AB - OBJECTIVES: The serine-protease inhibitor aprotinin protects platelet function during cardiopulmonary bypass. However, its safety and efficacy during deep hypothermic circulatory arrest (DHCA) is controversial, and aprotinin is suspected to cause thrombosis especially during hypothermia. The platelet GP IIb/IIIa inhibitor eptifibatide has been assumed to preserve platelet function during cardiopulmonary bypass without increasing bleeding complications. The aim of this study was to compare the effect of aprotinin and eptifibatide on platelet function under conditions of DHCA. METHODS: Heparinized blood from healthy volunteers (n = 10) was incubated in stasis for 30 minutes at 18 degrees C to simulate DHCA and compared to samples incubated at 37 degrees C. The effect of eptifibatide (2.5 microg/ml) and aprotinin (300 KIU/ml) on platelets under these conditions was analyzed by flow cytometry. Platelet aggregates were identified using CD41-antibody binding and size. GPIIb/IIIa function was evaluated with the activation-specific antibody PAC-1 after stimulation with 10 microM ADP. Aggregate numbers and antibody mean-fluorescence are reported as mean +/- standard deviation. RESULTS: Hypothermia induced a 2.5-fold increase of aggregates ( p < 0.001) and a 2.6-fold increase of GPIIb/IIIa activation ( p < 0.001). This effect was not influenced by aprotinin but almost completely inhibited by eptifibatide ( p < 0.001). CONCLUSIONS: Aprotinin has no procoagulatory effect on platelet function during hypothermia but is not protective either. Eptifibatide inhibits hypothermia-induced platelet aggregation in vitro and may prevent aggregate sequestration in the microvasculature and consecutive ischemic organ damage in vivo. PMID- 15786006 TI - Cardiothoracic surgery after heart and heart-lung transplantation. AB - OBJECTIVE: We sought to examine our management and the outcomes of cardiothoracic procedures after heart and heart lung transplantation. METHODS: We performed a retrospective review of cardiothoracic surgical procedures carried out between 1990 and 2004 in patients who had previously undergone heart or heart-lung transplantation at our institution. RESULTS: Twenty-one out of 340 patients (6.2 %) were identified. Cardiothoracic surgery was performed 44.4 +/- 33 months (range 1 - 115 months) after transplantation. Predominant types of surgery were coronary artery bypass grafting due to allograft vasculopathy (n = 5), aortic surgery due to acute dissection (n = 3), biventricular assist device implantation due to acute rejection (n = 1), tricuspid valve repair (n = 1), multiple cardiac surgical procedures including coronary artery bypass grafting, retransplantation, and tricuspid valve replacement (n = 2), explantation of a functionless heterotopic transplanted heart (n = 1). Lung surgery was performed in six patients due to pneumonia (n = 2), primary lung carcinoma (n = 3), lung torsion following heart-lung transplantation (n = 1). All patients underwent either lobectomy or segmental lung resection. Single lung retransplantation (n = 2) after prior heart-lung transplantation due to bronchiolitis obliterans was performed. In one patient a pneumonectomy (n = 1) due to severe chronic rejection of the contralateral lung was performed. Six subsequent deaths after cardiothoracic procedures were recorded after 1, 4, 78, 163, 205, and 730 days, respectively. Causes of death were advanced carcinoma (n = 1), multi-organ failure due to sepsis (n = 2), sudden heart death (n = 2), and advanced heart failure (n = 1). Fifteen out of 21 patients having undergone cardiothoracic procedures (71.4 %) survived the observation period of 56.6 +/- 34 months (range 1 - 114). CONCLUSIONS: Reasons for cardiothoracic procedures after prior heart or heart-lung transplantation were allograft vasculopathy, aortic dissections years after transplantation, chronic rejection, and either lung infections or malignancies. Surgical repair can be performed with an acceptable operative risk and good long-term survival rates. PMID- 15786007 TI - Patient attitudes to sternotomy and thoracotomy scars. AB - BACKGROUND: There are no data comparing patient attitudes to sternotomy and thoracotomy scars following surgery for congenital heart disease (CHD). METHODS: Two hundred and one patients with a scar from CHD surgery (105 sternotomy, 36 thoracotomy, and 60 both scars) had a structured interview to explore attitudes to their scar. RESULTS: Comparable proportions of each group reported that they did not like or hated their scar (23/105 [22 %] sternotomy, 9/36 [25 %] thoracotomy, 17/60 [28 %] both scars). Significantly more patients stated that they where embarrassed by and/or their choice of clothing was affected by a thoracotomy scar (20/36, 56 %) than those with a sternotomy scar (36/105, 34 %), p = 0.04. This was also seen when comparing sternotomy alone with both scars (36/105 [34 %] vs. 34/60 [57 %], p = 0.008). CONCLUSIONS: Adults who have undergone surgery for CHD are more likely to have a negative attitude to a thoracotomy than a sternotomy scar. Before a change in surgical approach is considered based on patient preferences, the acceptability and psychological impact of the different scars following surgery needs formal study. PMID- 15786008 TI - A low-flow adaptation phase improves shear-stress resistance of artificially seeded endothelial cells. AB - INTRODUCTION: The purpose of this study was to evaluate the effect of different adaptation phases on the shear-stress resistance of endothelial cells seeded artificially onto vascular prostheses and biological heart valves. MATERIAL AND METHODS: Human endothelial cells (EC), fibroblasts (FB), and smooth muscle cells (SMC) were isolated from vena saphena magna pieces and expanded in culture. Group A: 15 polyurethane vascular grafts (20 mm diameter) were seeded with FB and SMC (53 +/- 1.2 million cells), followed by EC seeding (39 +/- 0.9 million cells). Group B: eight stentless porcine valves (Freestyle, Medtronic, USA) were seeded with FB (68 +/- 1.5 million cells) and EC (42 +/- 1.1 million cells). Shear stress testing was done under pulsatile flow (pulse rate: 80 pulses/min.). Adaptation phase: flow was set to 0.9 +/- 0.3 l/min (systolic pressure: 40 - 50 mm Hg). High flow was 3.2 +/- 0.6 l/min. (systolic pressure: 140 - 160 mm Hg) and lasted over four hours in all groups. The vascular grafts were divided into three groups (n = 5 each): group 1 (high flow immediately), group 2 (adaptation phase of 15 minutes), and group 3 (adaptation phase of 30 minutes). The valves either were given high flow immediately (n = 4) or had an adaptation phase of 30 minutes (n = 4). Specimens were obtained after cell seeding, before, and after perfusion. RESULTS: A confluent EC layer was achieved on all grafts. After perfusion without adaptation, large defects within the cell layer were found. No FB and SMC were seen at the bottom of these defects. In group B, the defects were largest on the ventricular surface of the leaflets. After an adaptation phase of 15 minutes in group A, only a few defects within the EC layer were detected with a still confluent FB and SMC. After a 30-minute adaptation phase defects within the EC layer were very rare and no interruption of the underlying FB and SMC layer was seen. Immunohistochemical staining for factor VIII and CD31 proved the EC to be viable and staining for collagen IV and laminin revealed the formation of a basement membrane. After perfusion, the specimen also stained positive for eNOS. CONCLUSION: An adaptation phase of 30 minutes proved to be sufficient to allow artificially seeded endothelial cells to adapt to shear stress. The formation of a basement membrane was of great importance for the maintenance of a confluent EC layer. PMID- 15786009 TI - Effect of different operative techniques for myocardial revascularisation on hemodynamics and myocardial perfusion in a porcine model. AB - BACKGROUND: During surgical coronary revascularisation hemodynamics and myocardial contractility can be affected. This in vivo study aimed to determine the effects of different operative techniques on hemodynamics and regional myocardial perfusion. METHODS: In 24 pigs IMA to LAD bypass was constructed using ECC (n = 8) and cardioplegic arrest, OPCAB techniques (n = 8), or the Impella elect 100 support device (n = 8). 8 animals received a sham operation. Mean arterial pressure (MAP), cardiac output (CO), and left ventricular pressure (LVP, LVdp/dt) were recorded. Regional myocardial perfusion (RMP) of both ventricles was assessed by fluorescent microspheres. RESULTS: MAP significantly decreased during revascularisation in all groups ( p < 0.05), staying below preoperative values thereafter ( p < 0.05). After ECC norepinephrine was administered to maintain MAP. CO and LVdp/dt were impaired more distinctly during OPCAB than with Impella ( p < 0.05) during subsequent recovery. RMP showed global reactive hyperemia during early reperfusion after ECC, remained unchanged in OPCAB, and showed low flow during and after Impella pump run ( p < 0.05). CONCLUSIONS: ECC led to hemodynamic impairment with post-ischemic reactive hyperemia. OPCAB created hemodynamic depression but left RMP unchanged. Hemodynamic depression can be reduced by the Impella pump, however regional myocardial blood flow is decreased. PMID- 15786010 TI - Comparison of early postoperative results of thymectomy: partial sternotomy vs. videothoracoscopy. AB - BACKGROUND: The aim of this study was to compare the early postoperative results of thymectomy operations after partial sternotomy and videothoracoscopy for myasthenia gravis. METHODS: A total of 51 thymectomy operations were reviewed. The surgical procedure was simple thymectomy with partial sternotomy in the first 19 patients (Group I) and videothoracoscopic thymectomy (Group II) in the remaining patients. Both groups were compared in terms of preoperative data (age, gender, classification, duration of disease, medications), operative data (operation time, the mean amount of drainage, the duration of chest tube drainage), and postoperative data (duration of hospital stay, complications and pain). RESULTS: Groups were statistically uniform in terms of preoperative and operative data. Statistically significant differences were noted for the duration of chest tube drainage (48.8 vs. 29.8 hours, p < 0.001), the amount of drainage (264.4 vs. 178.6 ml, p = 0.001), the length of hospital stay (5.6 vs. 2.3 days, p = 0.000), and the visual analogue scale score (4.8 vs. 3.1, p < 0.001). CONCLUSIONS: Thymectomy with videothoracoscopic surgery demonstrated a more comfortable and faster recovery period without deterioration in myasthenic status. PMID- 15786011 TI - Frequency of loss of heterozygosity at 3 p, 9 p, 13 q, and 17 p is related to proliferative activity in smokers with stage I non-small cell lung cancer. AB - BACKGROUND: Tumor cells of lung cancer exhibit genetic abnormalities as well as high proliferative activity. The purpose of this study was to evaluate the relationship of genetic abnormalities and smoking status, histological type, and tumor proliferative activity in resected samples of stage I non-small cell lung cancer (NSCLC). METHODS: We evaluated 126 samples of stage I NSCLC from patients who underwent complete resection between 1988 and 1993. Loss of heterozygosity (LOH) was assessed using primers that amplified polymorphic microsatellite markers at D3S1300, D3S643, D3S1317, D9S171, IFNA, D13S153, and TP53. Expression of Ki-67 nuclear antigen was examined using immunohistochemical methods to assess tumor proliferative activity. RESULTS: The Fractional Regional Loss index (FRL) was significantly higher in squamous cell carcinoma samples than in adenocarcinoma samples (p < 0.0001). In smokers, Ki-67 labeling index (LI) in high-FRL cases was significantly higher than in low-FRL cases (p < 0.0001). CONCLUSION: The frequency of LOH at 3 p, 9 p, 13 q, and 17 p was related to proliferative activity in smokers with stage I non-small cell lung cancer. PMID- 15786012 TI - UFT inhibits lung metastases in spontaneous metastasis model of lung cancer. AB - BACKGROUND: UFT, an oral 5-fluorouracil derivative, is the only drug that is effective as a postoperative adjuvant therapy for non-small cell lung cancer (NSCLC) [ ], but the mechanism of the action remains unclear. We examined whether UFT and/or its metabolite, gamma-hydroxybutyric acid (GHB) inhibits lung metastases in a mouse model. METHODS: Lewis lung carcinoma cells were implanted into the foot pads of C57 BL/6 mice, and mice were treated with UFT or GHB. RESULTS: Both the mean number of metastatic nodules and the mean lung weight for UFT-treated mice (11.4 and 192.1 mg, respectively) were significantly lower than those for saline-treated mice (41.5 and 415.0 mg, respectively) (p < 0.001 for both). UFT did not inhibit tumor growth at the primary sites (foot pads). No significant body weight loss was documented in UFT-treated mice. GHB did not inhibit development of lung metastases even when a higher dose was used. CONCLUSIONS: UFT inhibits development of lung metastases without any toxicity in mouse model, which may explain the efficacy of postoperative administration of UFT for resected NSCLC. PMID- 15786013 TI - The importance of histology in the evaluation of pulmonary transplantation: carcinomatous lymphangitis. AB - Pulmonary carcinomatous lymphangitis is a form of neoplastic metastatic spread to the lungs, which represents a poor prognosis for the patient. The physician may be confronted by a differential diagnosis with diffuse pulmonary interstitial affections requiring specific treatment, including lung transplantation. We present the case of a patient, diagnosed with pulmonary interstitial disease with rapidly progressive worsening of lung function, who was considered for lung transplantation. Videothoracoscopic lung biopsy demonstrated the existence of carcinomatous lymphangitis, which completely changed the therapeutic direction. PMID- 15786014 TI - Intracavitary metastatic hepatocellular carcinoma of the right ventricle. AB - We report a rare case of direct intracavitary metastasis of a hepatocellular carcinoma (HCC) to the right ventricle (RV) without intravascular involvement. A 65-year-old female, with a history of HCC and partial right hepatectomy, developed symptoms of congestive heart failure. Echocardiography revealed a large tumor in the RV with extension to the outflow tract. Palliative excision of the cardiac tumor under cardiopulmonary bypass and cardioplegic arrest was performed due to severe hemodynamic compromise. The final pathology report disclosed metastatic HCC. Tumor recurrence in the RV was noted 2 months later and the patient succumbed to the disease. PMID- 15786015 TI - Living-donor lobar lung transplantation for pulmonary and abdominopelvic lymphangioleiomyomatosis. AB - We performed emergency living-donor lobar lung transplantation (LLTx) successfully in a 24-year-old woman with end-stage pulmonary and massive abdominopelvic lymphangioleiomyomatosis (LAM). Preoperatively, her respiratory condition was critical, but abdominopelvic lesions had been well controlled with medication. No LAM patients with massive abdominopelvic lesions as in the present case have been reportedly treated by LLTx previously. The present case demonstrates that LLTx can be a therapeutic option for end-stage pulmonary LAM with massive abdominopelvic involvement. PMID- 15786016 TI - Non-aneurysmal inflammation of the aorta in combination with aortic valve disease. AB - A 61-year-old man presented with aortic valve stenosis, coronary artery disease in combination with rheumatic arthritis, peri-aortic inflammation and fibrosis with aortitis. In the pathological literature this has been termed chronic periaortitis, while in the surgical literature, non-aneurysmal dilatation of the aorta associated with atherosclerosis and inflammation has been described as aortitis. The name chronic periarteritis refers to the associated involvement of coronary arteries. Although different terminologies have been used, it is suggested that all of them most likely represent the same disease entity. PMID- 15786020 TI - Lymphoma: diagnosis, staging, natural history, and treatment strategies. AB - Non-Hodgkin's lymphoma (NHL) is not a single disease, but a group of closely related B- and T-cell cancers of the lymphatic system. The incidence of NHL is rising, particularly in the countries of the industrialized world. The increased incidence is poorly understood, but several risk factors have been postulated to be associated with NHL, including: exposure to chemicals, viral infections, organ transplantation and blood transfusion, family history, and lifestyle factors. Precise staging of NHL is a prerequisite for the selection of a suitable therapeutic regimen and influences the likelihood of its success. Staging of lymphoma is traditionally conducted using tumor biopsy, imaging (X-ray, computerized tomography [CT], magnetic resonance imaging, lymphangiogram, gallium scan using 67 Ga citrate single photon emission CT [ 67 Ga-SPECT], or positron emission tomography), blood tests, bone marrow examination, and examination of cerebrospinal fluid. CT is the most commonly used imaging technique, but several studies indicate that other techniques, such as 67 Ga-SPECT, are more sensitive and better predictors of response. Low- and high-grade lymphomas differ markedly in prognosis and response to treatment. The management of NHL has been characterized by the increasing recognition that distinct subgroups of NHL respond differently to various therapeutic approaches. In follicular lymphoma (FL), a successful approach has been to combine fludarabine with mitoxantrone (FM), resulting in an overall response rate of 89% (67% complete remission). In an ongoing phase III trial in patients with untreated, advanced, low-grade follicular lymphoma, FM was compared with CHOP (cyclophosphamide/doxorubicin/vincristine/prednisone) with or without rituximab. Patients previously treated with FM achieved a significantly better complete remission rate (67% v 38%; P = .0013) and a better molecular remission (36% v 20%; P = .049) than patients previously treated with CHOP. Following rituximab treatment, 88% of patients who had received FM achieved clinical and molecular remission compared with 70% who had received CHOP. Strategies using various chemotherapy combinations, including innovative agents such as 90 Y-ibritumomab tiuxetan, show promise in the treatment of NHL, particularly indolent NHL, and hopefully will lead to an improvement in prognosis. PMID- 15786018 TI - Nonparametric tests of association of multiple genes with human disease. AB - The genetic basis of many common human diseases is expected to be highly heterogeneous, with multiple causative loci and multiple alleles at some of the causative loci. Analyzing the association of disease with one genetic marker at a time can have weak power, because of relatively small genetic effects and the need to correct for multiple testing. Testing the simultaneous effects of multiple markers by multivariate statistics might improve power, but they too will not be very powerful when there are many markers, because of the many degrees of freedom. To overcome some of the limitations of current statistical methods for case-control studies of candidate genes, we develop a new class of nonparametric statistics that can simultaneously test the association of multiple markers with disease, with only a single degree of freedom. Our approach, which is based on U-statistics, first measures a score over all markers for pairs of subjects and then compares the averages of these scores between cases and controls. Genetic scoring for a pair of subjects is measured by a "kernel" function, which we allow to be fairly general. However, we provide guidelines on how to choose a kernel for different types of genetic effects. Our global statistic has the advantage of having only one degree of freedom and achieves its greatest power advantage when the contrasts of average genotype scores between cases and controls are in the same direction across multiple markers. Simulations illustrate that our proposed methods have the anticipated type I-error rate and that they can be more powerful than standard methods. Application of our methods to a study of candidate genes for prostate cancer illustrates their potential merits, and offers guidelines for interpretation. PMID- 15786021 TI - Imaging: staging and evaluation of lymphoma using nuclear medicine. AB - The management of lymphoma is dependent on accurate staging of the disease and evaluation of histology and other risk factors. Advances in imaging techniques have improved the assessment of disease status and evaluation of the efficacy of different treatment modalities. While computed tomography remains the cornerstone of imaging for the assessment of disease status, it provides no understanding of the metabolic or functional parameters of the disease. Nuclear medicine techniques permit the evaluation of functional status, and nuclear medicine is likely to have its greatest impact in the detection of viable tumor in persistent masses. Nuclear imaging can be conducted using single photon agents, such as 67 Ga-citrate with SPECT (single photon emission computed tomography), or with positron emitters, such as 18 F-fluorodeoxyglucose (FDG) with PET (positron emission tomography). Many studies have indicated that FDG-PET is as good as or better than 67 Ga-SPECT for the detection of lymphoma. Thus, FDG-PET may be preferable to 67 Ga-SPECT for disease evaluation, staging, and follow-up. The superiority of FDG-PET compared with other imaging modalities strongly argues that it should be incorporated into the treatment paradigm. At present, FDG-PET scanning is not routinely available in all institutions; however, a role can be indicated for FDG-PET in several areas of lymphoma management, including initial staging, predicting response to therapy (during and following chemotherapy), and identification of residual tumor. This article examines the role of the different imaging techniques available and the use of these techniques in the staging and evaluation of patients with Hodgkin's and non-Hodgkin's lymphoma. PMID- 15786022 TI - Concepts in radiotherapy and immunotherapy: anti-CD20 mechanisms of action and targets. AB - Immunotherapy became a feasible therapeutic approach following the development of monoclonal antibody technology. Despite many small clinical trials using a wide variety of antibodies, in hematologic malignancies, success has largely been restricted to a few antibodies generated against antigens expressed on the tumor cell surface: rituximab (anti-CD20) and alemtuzumab (anti-CD52) are the most widely used monoclonal antibodies. CD20 is expressed on B cells, and rituximab has been widely used in the treatment of various histologies of B-cell non Hodgkin's lymphoma. The mode of action of rituximab is discussed in this article. Despite extensive empiric clinical trial experience, the critical factors important in the mechanism of tumor cell kill by monoclonal antibodies continue to be elusive and the subject of debate. The major immune mechanisms of action of rituximab include complement-dependent cytotoxicity and antibody-dependent cellular cytotoxicity. While some investigations have suggested that the complement system is essential to tumor cell kill with rituximab; most lines of evidence point to the importance of antibody-dependent cellular cytotoxicity. Fc receptor binding appears to be critical in determining efficacy. Observations from several studies have shown that the response rate to single-agent rituximab is better in patients who have higher affinity polymorphisms in their Fc receptors. The other mechanisms that may play a role in rituximab therapy include direct anti-tumor effects mediated by the antibody binding to cell-surface CD20 antigen. Although this is more controversial, observations in tumor cell lines following CD20 ligation suggest that direct effects may be important. PMID- 15786023 TI - Concepts in radioimmunotherapy and immunotherapy: Radioimmunotherapy from a Lym-1 perspective. AB - Conventional chemotherapy regimens cure fewer than 50% of patients with aggressive non-Hodgkin's lymphoma, and fewer than 5% of patients with indolent lymphomas. However, the majority of patients remain responsive to remarkably low doses of external beam radiotherapy. A logical strategy for the treatment of non Hodgkin's lymphoma is radioimmunotherapy (RIT); systemic radiation targeted to tumor cells using monoclonal antibodies. RIT involves continuous exposure to low dose-rate radiation, with the intensity of the dose decreasing over time, and as such is distinct from conventional radiotherapy and chemotherapy. RIT has several advantages over monoclonal antibody therapy. For example, a functional immune system is not an absolute requirement to kill tumor cells, and, depending on the radiolabel used, beta-emissions are effective over 100 to 500 cell diameters, resulting in a crossfire effect on nearby tumor cells. The crossfire effect enables the eradication of cells that are not necessarily targeted by the antibody, but are affected by the radiation. The success of RIT depends on which antibody and radioisotope is used. This article examines how the antibody, radioisotope, chelator, and linker affect the safety and efficacy of RIT. The different approaches to dosing are also considered. PMID- 15786024 TI - Use of 90Y-ibritumomab tiuxetan in non-Hodgkin's lymphoma. AB - The increase in the incidence of non-Hodgkin's lymphoma (NHL) that has occurred over recent decades is expected to continue. Therapeutic options for patients with NHL have improved over the past 20 years, but almost all patients with low grade lymphoma and approximately 50% of patients with high-grade lymphoma eventually die of their disease, regardless of the regimen used. Thus, there is a continuing need for novel therapeutic options. One such strategy is targeted radioimmunotherapy, which is an attractive approach because lymphoma cells are inherently sensitive to radiation. 90 Y-ibritumomab tiuxetan (Zevalin; Biogen Idec Inc, San Diego, CA, and Schering AG, Berlin, Germany) was the first radioimmunotherapy agent to be approved by the US Food and Drug Administration for the treatment of patients with relapsed, low-grade B-cell NHL. 90Y ibritumomab tiuxetan comprises the murine IgG1 anti-CD20 antibody ibritumomab, covalently linked to the beta-emitter 90 Y by a chelator tiuxetan. A prospective trial comparing 90Y-ibritumomab tiuxetan with single-agent rituximab showed an overall response rate (ORR) to 90Y-ibritumomab tiuxetan of 80% (34% complete response [CR]/unconfirmed CR [CRu]) compared with 56% (20% CR/CRu) with rituximab (P = .002). Of patients achieving a CR/CRu, 32% were still in remission at 3 to 4 years of follow-up. Similar efficacy (83% ORR, 43% CR/CRu) has been reported with 90 Y-ibritumomab tiuxetan in patients with relapsed or refractory low-grade NHL with mild thrombocytopenia (platelet counts 100,000 to 149,000/mm3 ), and in patients with rituximab-refractory NHL (ORR 74% [CR 15%] compared with an ORR 32% to last rituximab treatment). Safety data compiled from patients entered into five studies have confirmed initial observations that the toxicities encountered with 90Y-ibritumomab tiuxetan therapy are mainly hematologic and transient. As part of a consolidated clinical approach to the ongoing development of 90Y ibritumomab tiuxetan, studies are currently being conducted in the United States and Europe to examine the role of this agent in first-line therapy of indolent NHL, in diffuse large B-cell lymphoma, and in combination with chemotherapy with peripheral blood stem cell support. PMID- 15786025 TI - 90Y-ibritumomab tiuxetan: rationale for patient selection in the treatment of indolent non-Hodgkin's lymphoma. AB - Radioimmunotherapy with 90Y-ibritumomab tiuxetan (Zevalin; Biogen Idec Inc, San Diego, CA, and Schering AG, Berlin, Germany) was approved in the United States in 2002 for patients with relapsed or refractory, low-grade, follicular, or transformed B-cell non-Hodgkin's lymphoma, including patients with rituximab refractory disease, and in Europe in 2003. This agent has yielded good results in the treatment of patients with relapsed or refractory non-Hodgkin's lymphoma, for whom limited treatment options are available. Radioimmunotherapy with 90Y ibritumomab tiuxetan is well tolerated and produces statistically and clinically significant higher overall and complete response rates than rituximab alone. Furthermore, treatment can be safely administered on an outpatient basis, with minimal disruption to patients' daily routines. The 90Y-ibritumomab tiuxetan treatment regimen, appropriate patient selection, and risk assessment are discussed in this article. PMID- 15786026 TI - Development of 131I-tositumomab. AB - The median survival for patients with advanced indolent non-Hodgkin's lymphoma (NHL) has remained at 7 to 8 years since the 1960s. Targeted treatment using radioimmunotherapy (RIT), radiolabeled monoclonal antibodies directed against tumor-specific antigens, is an attractive option for this patient population, combining the advantages of an active biologic therapy with low dose-rate irradiation of an inherently radiosensitive tumor. Two anti-CD20 RIT agents have now been approved for the treatment of refractory NHL: 90Y-ibritumomab tiuxetan (Zevalin; Biogen Idec Inc, San Diego, CA, and Schering AG, Berlin, Germany) is approved in both the United States and Europe, and 131I-tositumomab (Bexxar; Corixa Corp, Seattle, WA) is approved only in the United States. This article discusses the development of 131I-tositumomab. Because 131I-labeled antibody clearance varies significantly among patients, prescription of 131I-tositumomab activity must be based on a calculated total-body dose derived from quantitative whole-body imaging. The maximum tolerated total-body dose has been established at 75 cGy in patients with adequate bone marrow reserves and less than 25% bone marrow involvement by lymphoma (65 cGy in patients with mild thrombocytopenia; 45 cGy in patients who have received stem cell transplantation). In a phase III trial, overall response rate (ORR) and complete response (CR) rate were significantly higher following 131I-tositumomab than following the patient's last qualifying chemotherapy (ORR, 65% v 28%; P <.001; CR, 20% v 3%; P <.001). 131I tositumomab has also been shown to be effective in patients who are refractory to rituximab (ORR, 70%; CR, 32%) and as first-line therapy in patients with NHL (ORR, 97%; CR, 63%). The major side effects of 131I-tositumomab are hematologic. In the phase III study, 20% of patients experienced grade 4 neutropenia and 22% experienced grade 4 thrombocytopenia. Myelodysplastic syndromes or secondary acute myeloid leukemia have been reported in 8.4% of patients with chemotherapy refractory disease treated with 131I-tositumomab, but have not been observed to date in patients receiving 131I-tositumomab as first-line therapy. Future progress in NHL management is likely to include RIT as part of a multi-modality approach; trials are planned or currently underway to investigate the combination of RIT with chemotherapy regimens. PMID- 15786027 TI - Future trends in radioimmunotherapy. AB - Radioimmunotherapy (RIT) is currently approved in the United States only for use in low-grade follicular or transformed non-Hodgkin's lymphoma. One of the major issues in the construction of new clinical trials is deciding in which phase of disease RIT should be applied: upfront, as part of first-line treatment, in relapsed/refractory patients, in bulky or 'minimal residual disease', or in patients failing 'cold' antibody treatment? An important consideration for future trends in RIT is how the efficacy of RIT can be increased, while limiting toxicity. To increase efficacy, it is important to understand the reasons why RIT can fail in some patients. Efficacy might be increased by improving tumor to normal organ ratios of antibody (using synthetic clearing agents to clear nontumor-bound antibody from the bloodstream), or by combining RIT with radiosensitizers, such as fludarabine. Methods of reducing toxicity are also being investigated (eg, by splitting the full dose of RIT, or using biological prognostic factors to identify patients who are likely to experience myelosuppression). Biological prognostic factors may also predict disease response and can be used to stratify patients into risk groups at diagnosis, which may predict outcome according to a limited set of prognostic factors. It is also important to establish the effect of prior chemotherapy regimens on RIT, and likewise to determine to what extent RIT compromises subsequent chemotherapy treatment. This article attempts to address these issues and anticipate future trends in RIT. PMID- 15786028 TI - The future of radioimmunotherapy: a PET perspective. AB - The future success of radioimmunotherapy (RIT) lies in building relationships between oncologists and nuclear medicine physicians. Communication between these specialists is key to the progress of radioimmunotherapy, and a team approach is required among all those involved in the process, not only for the benefit of the patient, but also for the success of research endeavors. Identification of the key questions regarding diagnosis and management of cancer must be formulated to design appropriate clinical trials incorporating radioimmunotherapy. The use of imaging modalities to conduct radiotracer studies may provide valuable information with regard to drug delivery and target specificity. One such approach is positron emission tomography (PET), a functional imaging modality with high sensitivity and specificity for tumor detection. The recent addition of hybrid PET/computed tomography, enabling biologic and anatomic whole-body imaging to be performed in one examination, has further facilitated the assignment of biologic abnormalities to anatomic structures. The most widely used radiotracer for PET is 18 F-fluorodeoxyglucose; however, fluorodeoxyglucose is a relatively nonspecific tracer. Novel, more specific tracers are being designed to assess cell proliferation. Radiolabeled non-catabolized derivatives of thymidine that are incorporated into cellular DNA are undergoing clinical trials for diagnostic efficacy using PET. In vivo anatomic micro-imaging technologies, such as microPET, microcomputed tomography, and micro-magnetic resonance imaging, enable the assessment of metabolism, cell communication, and gene expression in animal models. Fast-tracking imaging methods through translational research to human use is critical if imaging is to aid in the delivery of optimal cancer treatments. This article presents an overview of the way in which imaging may facilitate the evaluation of new treatment modalities to advance patient treatment and care. PMID- 15786374 TI - [A nephrologist's tasks in nephrolithiasis]. AB - The epidemiological impact of nephrolithiasis stems from a significant and increasing prevalence in western countries. While the kidney is the end-organ of the disease, the causes are often more general, including metabolic derangements, pri-mary diseases of other organs and systems, hereditary renal or non-renal defects. In this context, nephrological expertise is highly recommended and could considerably improve disease outcomes. The nephrologist's involvement should start while the patient is acutely affected by renal colic. In this setting medical intervention is aimed at counteracting pain, favoring progression in the urinary tract, and preventing renal injury. The choice for urological procedures should take into account the potential for harmful effects of obstruction, infection, and prolonged pain. After the patient has undergone non-invasive procedures medical intervention improves the management of residual fragments, reduces the risk of stone recurrence, and increases compliance to the stone center, as a premise for considering patients for metabolic evaluation and subsequent medical treatment. Current study protocols, including chemistries, physicochemistry and possibly genetics, are the basis for a rational treatment of recurrent stone disease. Secondary nephrolithiasis caused by systemic disorders is screened out and treated specifically. Hereditary forms can be identified by genetic analysis and strictly followed and treated. While medical therapy can cure some types of renal stones, prolonged remission is seldom obtained in calcium nephrolithiasis. However, recurrence rates are greatly reduced, and this lessens the need for urological procedures, risk of infection/obstruction and, ultimately, progression to renal insufficiency. In the face of a multidisciplinary approach to renal stone disease, the nephrologist has a key role in the successful management of these patients. PMID- 15786373 TI - [New immunodepressant drugs for the prevention and control of kidney transplant rejection]. AB - The increasing number of kidney transplantations performed yearly in Italy should prompt nephrologists to improve their knowledge in these patients that fall "by default" in the nephrologists' province. In taking on of this task in addition to a skilled clinical and organizational capacity, it demands experience and an updated knowledge of the immunosuppressive drug machinery currently available. Calcineurin inhibitors (cyclosporine and tacrolimus) are well-defined milestones, currently used. Less well known are some recent drugs such as rapamycin and its analogues, mycophenolate mofetil and mycophenolic acid, anti-CD 25 monoclonal antibodies, chimeric and humanized. These drugs are, or they have become, available commercially in the last few years or they are going to be registered in the near future. However, their registration is due to years of usage in the large majority of current immunosuppressive regimens where they can play the role of a primary or an ancillary drug. Other drugs such as FH778, FTY720, anti-CD 20 (rituximab) and anti-CD 52 (alentuzimab) monoclonal antibodies are still undergoing trials in many collaborative phase 2 and 3, studies. Finally, a third group of drugs is targeted at the co-stimulator molecule network and it is an interesting and challenging opportunity for the near future. A detailed knowledge of the overall therapeutic armamentarium is useful in the kidney transplantation field where all drugs, from the beginning to date, must always be kept in mind. The new drugs do not supplant the old drugs. Knowing the pros and cons of each immunosuppressive drug used in the therapeutic management of kidney transplantation is crucial in improving the clinical results in general, and in tailoring the best regimens in different clinical settings pro-posed by kidney recipients today. PMID- 15786375 TI - [Metabolic acidosis in patients with chronic kidney diseases: why and when to treat it?]. AB - Metabolic acidosis is a common complication in patients with advanced chronic renal diseases and dialytic treatments are unable to correct it completely. In hemodialysis (HD) patients, severe metabolic acidosis is associated with an increased risk of death. Evidence from several experimental studies suggests that even mild metabolic acidosis is associated with systemic effects. Acidosis is implicated in endocrine changes and has negative repercussions on bone and protein metabolism. In addition, recent observations suggest that acidosis triggers inflammation and accelerates the progression of chronic kidney diseases. As a contradictory finding, acidosis can reduce circulating leptin. Clinical studies on the nutritional effects of metabolic acidosis correction have shown mildly favorable effects. Taking into account the systemic effects of metabolic acidosis it is suggested that even mild metabolic acidosis is corrected. However, the new findings concerning the systemic effects of acidosis must be evaluated in controlled trials. PMID- 15786376 TI - [Regional clinical audit, guideline targets, and local and regional benchmarks]. AB - Regional clinical Audit, guideline Targets and local and regional Benchmarks In order to improve the quality of dialysis treatment, we have devised some routines, particularly suitable for electronic data management systems. First, we suggest a systematic monthly analysis of 10 common clinical performance measures (CPM), with the following guideline based targets: predialysis systolic blood pressure (SBP) < 140 mmHg; session length >/= 240 min; dialysis dose (spKt/V) >/=1.3; normalized protein catabolic rate (NPCR) >/=1.2 g/kg/d; hemoglobin (Hb) >/=11 g/dL; serum calcium (Ca) 8.4-9.5 mg/dL; serum phosphorus (P) 3.5-5.5 mg/dL; Ca x P /=20 mmol/L; serum potassium (K) 3.5-6.0 mmol/L. The Hb target should be reached in at least 85% of all maintenance hemodialysis (HD) patients in the unit; for all other targets, an arbitrary >/=80% is proposed. Since the above percentages are quite difficult to reach on a short-term basis, an intermediate local or regional standard (benchmark) could be devised as an average of the percentage of patients who actually reach the targets for each CPM at any dialysis unit in a given regional area; and therefore, from truly comparable patients. As an example, we simulated a regional audit by using the above targets with available data from 398 patients from southern Italy. A further step in this process was to find the cause(s) of failure in each patient who did not reach the targets. To this end, we suggest a systematic search of the well-known factors that could affect each CPM, for each failed patient. As an example, we screened all patients with Hb < 11 g/dL at a single unit, to establish the presence/absence of any common cause associated with inadequate response to epoetin treatment. Moreover, by using criteria for prescribing iron therapy or increasing epoetin dose, we found that some patients did not receive the appropriate therapy after blood sampling results. To avoid this possible common problem, we suggest the need for a monthly report of failure cases for any particular CPM and a check that the appropriate treatment has been delivered to all patients at the dialysis unit. This should also favor guideline implementation. PMID- 15786377 TI - [Artificial intelligence--the knowledge base applied to nephrology]. AB - The idea that efficacy efficiency, and quality in medicine could not be reached without sorting the huge knowledge of medical and nursing science is very common. Engineers and computer scientists have developed medical software with great prospects for success, but currently these software applications are not so useful in clinical practice. The medical doctor and the trained nurse live the 'information age' in many daily activities, but the main benefits are not so widespread in working activities. Artificial intelligence and, particularly, export systems charm health staff because of their potential. The first part of this paper summarizes the characteristics of 'weak artificial intelligence' and of expert systems important in clinical practice. The second part discusses medical doctors' requirements and the current nephrologic knowledge bases available for artificial intelligence development. PMID- 15786378 TI - [A 'lucky' patient]. AB - A 72-year-old male diabetic patient admitted to our operative unit of nephrology and dialysis underwent hemodialytic treatment because of rapidly progressive renal failure. A moderate hypertensive state was associated to nephrotic proteinuria and microematuria. Renal angiography showed a severe stenosis of the right renal artery and a smaller left kidney. Right renal artery stenting induced a significant reduction in serum creatinine (Cr) and the patient discontinued with the dialytic treatment. PMID- 15786379 TI - [On the mailing list: use of acyclovir in patients on hemodialysis]. AB - In the SIN Mailing List a message published in May 2004 gave rise to an interesting debate as to the use and dosage of acyclovir in dialysis patients for the treatment of varicella-zoster infection, considering the side effects (mainly neurological) reported in patients affected by renal insufficiency. In this issue, we summarised the main pharmacological characteristics of acyclovir and the clinical indications for using this drug. Finally, we analysed the literature data concerning acyclovir side effects in the setting of renal insufficiency. As they indicate that acyclovir-induced neurotoxicity in renal insufficiency patients may be unpredictable even at reduced dosages, this should discourage the use of acyclovir when severe renal insufficiency is present, unless severe herpes infection is present and after evaluating carefully the risk-benefit. In this case, the patient has to be kept under strict medical control to detect the acyclovir-induced side effects early. The use of new antiviral drugs, such as famciclovir or brivudine, to treat herpes virus infections in patients with renal insufficiency is currently not supported by sufficient data for a critical review. PMID- 15786380 TI - [Protocols and procedures]. AB - Evidence-based nursing increases behavior uniformity during everyday activities and in critical or complex situations. A recorded and programmed activity, the foreseeing of resources, the checking of application modalities and outcome evolution, and warning of complications, are the basis for a good pre-ordered flow chart. Nurses themselves build valuable protocols for every care procedure. This study describes central venous catheter (CVC) care as an example of a useful procedure. PMID- 15786381 TI - [Hemodiafiltration with endogenous reinfusion (HFR): evolution of the method]. AB - During hemodiafiltration (HDF) the increase in the ultrafiltration (uf) rate improves solute convective clearance. Therefore, considerable amounts of reinfusion liquid are necessary, whose quality must be guaranteed. The use of bags or bottles manufactured by industry causes many problems concerning handling (storage, repeated connections) and costs, and last but not least, it exposes the circuits to a contamination risk. Therefore, the technological research into on line production systems of sterile and ultrapure reinfusion solutions is justified. The increasing interest in the on-line production of reinfusion fluids from the dialysing solution dates back to the 1990s, and the data in the literature are statistically positive concerning the improvement in depurative performances connected to the uf increase. Although, to be objective, a problem still exists, in real-time the absolute guarantee of the sterility and apirogenicity of the solutions produced on-line is impossible. Using a two chamber filter, it is possible to produce reinfusion fluid from the uf of the patient himself, which has been 'regenerated' by a sorbent bed, in a closed circuit. This action eliminates any sterility problems, whilst, in addition, providing the possibility for good substance reinfusion such bicarbonates and essential and branched-chain aminoacids. This HDF method, hemo-filtrate reinfusion (HFR), has clinically demonstrated, beyond its ease of execution, a reduction in physiological component loss, a high biocompatibility and an overall action in contrast to MIA syndrome factors, i.e. malnutrition, inflammation and atherosclerosis. PMID- 15786382 TI - [Thermal sensor and on-line hemodiafiltration]. AB - Among the advantages of hemodiafiltration (HDF) cardiovascular stability seems the most popular. For several years the theory was that fluid infusion at a lower temperature induced similar effects to 'cool dialysis'. Due to the studies of Maggiore in 1982, we know that hemodialytic treatment per se induces in the majority of patients a rise in body temperature, on average 1.5-2 degrees C; the reasons for this phenomenon are unclear, even if theories developed in different trials take the following deputed to body cooling into account; an increase of the catabolism, linked to the bioincompatibility of the materials or to pyrogenic and cytokine-inducing substances, or a reduction in thermal scattering by the vasoconstriction of vasular districts such as skin and muscles. A device has been developed, the BTM (R) , which is able to measure the blood temperature at the arterial and venous side of the extracorporeal line and to modify the dialysate temperature, thereby inducing an analogous variation of the venous blood temperature. Therefore, we were able to precisely calculate the thermal balance during the dialytic treatment and work automatically to ensure sessions in which the body temperature remained stable, continuously (isothermic dialysis), or to observe that the thermal balance could vary during the same session (thermoneutral dialysis). Several multicentric studies, carried out in Europe, and particularly in Italy, have demonstrated that isothermic dialysis reduces the number of dialytic sessions complicated by symptomatic hypotension by 50%, particularly in hypotension prone patients. PMID- 15786383 TI - [Acetate free biofiltration (AFB) and ionic dialysance]. AB - Among various methods and models to evaluate the dialysis dose, the on-line measure of ionic dialysance gives good results. Compared to measured KT/V, ionic dialysance does not require blood sampling and, consequently, it reduces the risks and the costs. Functional evaluations during acetate free biofiltration (AFB treatments) confirm the usefulness of the on-line continuous measure by biosensors, especially when they are integrated in the dialysis machinery. PMID- 15786384 TI - [Integrated local management regarding patients suffering from the initial stage of chronic renal failure: NephroCare group pilot experience]. AB - BACKGROUND: Early diagnosis of the initial stage of chronic renal failure (CRF) is a highly important factor for reducing morbidity and mortality in patients suffering from this pathology. It delays their undergoing hemodialysis (HD) treatment and prevents its complications. This is a well known problem, and the wrong approach causes the so-called late referral phenomenon. Undoubtedly general practitioners (GPs), can, first, identify at an early stage, patients suffering from this syndrome, as they are the first point of contact for patients at risk (for example, diabetic and hypertensive patients). Furthermore, patients very often turn to their GP for various reasons, even those who are 'apparently in good conditions of health'. This study aimed to implement, through NephroCare and the Cooperatives of General Practitioners, 'Progetto Leonardo' and 'Cumae', the evidence that local cooperation between GPs and nephrologists can be effective. METHODS: In a localized geographical area (Pozzuoli, Bacoli-Monte di Procida), NephroCare organized meetings with GPs, and doctors belonging to the cooperatives, aimed at creating interaction and discussion concerning the etiology and pathogenesis of kidney diseases that lead to CRF, its complications, prevention and management. Screening of 46277 patients, with a 2-yr follow-up was carried out to identify the two categories at risk, diabetics and hypertensives with creatinine (Cr) >1.5 mg/dL. RESULTS: The screening identified among 7403 hypertensives and 2501 diabetics two groups of, respectively, 274 and 192 patients with Cr >1.5 mg/dL. All patients screened were invited to talk to a nephrologist, who shared, together with the GP, the decision concerning diagnosis and care. CONCLUSION: The outcomes of the project highlighted the importance of cooperation between GPs and nephrologists to implement the integrated management of patients, increasingly elderly, suffering from CRF. This was achieved because the nephrologists belonged to the same regional area as the patients and were thoroughly dedicated to the project. In times of reduced economic resources, a possible correct approach is facing the 'late referral' problem and, consequently, easier and better management of patient care and cost rationalization can be solved by the intervention of nephrologists working in local clinics. In the future, this will always guarantee continuous care for patients suffering from this pathology. PMID- 15786385 TI - [Early kidney-pancreas transplantation in type 1 diabetics prevents hemodialysis]. AB - BACKGROUND: The life-expectancy of type 1 diabetics (T1D) on dialysis is still shorter than that of non-diabetics. Pancreas transplantation (PT) in its different modalities should be considered as a life-saving procedure. METHODS: We analyzed our referral activity of T1D to PT from 1992. Since 2002, we have created a kidney and diabetes out-patient clinic devoted to the prevention of diabetic nephropathy and to the early referral of suitable T1D to combined kidney pancreas transplantation (KPT) and isolated pancreas (PTA). RESULTS: In the last 14 yrs, 25 T1D underwent KP in our district (620000 inhabitants). At the beginning, KPT was performed abroad, but then the borders were closed. After stopping in the mid 1990s, KP activity restarted addressing preemptive KPT and PTA. Currently, only one patient is on dialysis while awaiting KPT. Four T1D were evaluated and excluded from the list on medical grounds; two patients are on the list and a further two patients are currently under evaluation. CONCLUSIONS: The implementation of a cooperative network among dialysis and transplant centers, supported by devoted out-patient clinics allowed the effective prevention of the dialysis requirement in T1D. Out-patient clinics devoted to diabetic nephropathy should play a pro-active role in preemptive KP, including the 'new' option of islet transplantation according to the Edmonton protocol. PMID- 15786386 TI - [Epidemiology of chronic nephropathies in the referring population of a nephrology out-patient department]. AB - BACKGROUND: Data on chronic nephropathies at the starting of the dialysis (Renal Regitries) don't show the real frequency of these diseases, because they refer to a population, which has been previously selected by the course of the illness; there are a few and fragmentary data referred to the general population, observed in primary care or outpatients' departments. The aim of the present study is to measure the frequency of the chronic nephropathies observed in a nephrology outpatients'department and compare it with the frequency reported by other studies on local epidemiology. METHODS: We deal with the population referring to the nephrology outpatient's department in the Barletta Hospital during 2003 for 159.803 residents in all. The personal data have been analyzed, as well as the frequency of Chronic Renal Failure (CRF, 2 degrees -5 degrees stage MDRD), Hypertension and Diabetes. Our data have been compared to data reported by a local multicentric study performed in Basilicata in 2001. RESULTS: The main features of the analyzed population were: mean age 64.9 years and mean GFR 50,4 mL/m'/1,73 m2; there where more women than men, and 89.8% of patients had CRF, 67.2% Hypertension, 12.7% diabetes. The total referral to nephrologist was 242.8/100.000 residents. Our incident population significantly differs from the one of the Lucania, due to the inverted ratio of men to women, the higher degree and the earlier referral of CRF, with no significant difference in mean age. The percentage of patients with CRF and Hypertension was alike, whereas that of the diabetic patients was significantly lower. CONCLUSIONS: Although the outpatients' populations are neighbouring, they may significantly differ in features and frequency of chronic nephropathies. This phenomenon is the expression of referral factors, and demonstrates the importance of local epidemiology in appropriate care planning. PMID- 15786387 TI - [Valaciclovir-related neuro- and nephro-toxicity in two geriatric patients]. AB - BACKGROUND: Valaciclovir (VAL) is the L-valyl ester of acyclovir and it is quickly hydrolysed to acyclovir. Acyclovir-related nephrotoxicity is associated with high drug infusion doses and VAL-related neurotoxicity to pre-existing renal failure. We observed two geriatric patients with normal serum creatinine who developed acute renal failure (ARF) and neurotoxicity after ingesting the drug at the conventional therapeutic dose. CASE REPORTS: The first patient was a 77-year old female admitted to the intensive care unit because of anuria associated with coma and respiratory insufficiency. Baseline serum creatinine (Scr) was 0.9 mg/dL. Four days previously she had started VAL 1 g/bd for cutaneous HZ. Scr was 7 mg/dL and urinary output was <100 cc/24 h. The patient was ventilated and hemodialysis (HD) was started. After 6 days her urinary output increased and dialysis was discontinued. Two weeks after admission, Scr was 1.5 mg/dL, ventilation was discontinued and her consciousness level was normal. The second patient was a 73-year-old male. Baseline Scr was 0.7 mg/dL. He started VAL 1 g/TD because of hepatic cheratitis. After 4 days, he developed oliguria and stupor. At admission, Scr was 5.5 mg/dL, potassium was 6 mEq/L and the patient was unconscious. After drug withdrawal the diuresis increased and his consciousness level returned to normal. After 7 days Scr was 0.9 mg/dL. COMMENT: The clinical evolution suggested that the syndrome was related to the antiviral therapy. ARF preceded neurotoxicity. We suggest that high bioavailability, reduced glomerular filtration rate (GFR) related to age, and possibly the simultaneous ingestion of other drugs could have caused these side effects. We recommend the emphasis on potential side effects of antiviral therapy when treating geriatric patients even if Scr is in the normal range. PMID- 15786388 TI - [Hematological and iron parameters to predict mortality in ESRD]. AB - The prognostic value of hematological parameters other than hemoglobin (Hb) has been seldom investigated in hemodialysis (HD) patients. We used the predialytic assessment of blood ferritin, blood transferrin, transferrin saturation, blood iron, total iron binding capacity (TIBC), Hb, reticulocyte count, IRF, mean corpuscular volume (MCV), RDW, CHR, in our HD patients, as well as the weekly iron and erythropoietin (EPO) supplementation, to evaluate the relationship with death in the subsequent 12-month period. Data were divided into two groups (group M for dead patients and group V for patients remaining alive after 12 months) and mean +/- SD with significant differences (Student's t-test) were calculated. The following results were obtained: blood transferrin: M (n=21) 1.78 +/- 0.57, V (n=96) 1.72 +/- 0.36 g/L (p=ns); blood iron: M (n=22) 8.66 +/- 5.07, V (n=97) 10.50 +/- 4.57 mmol/L (p=ns); TIBC: M (n=21) 42.69 +/- 13.63, V (n=98) 40.36 +/- 10.33 mmol of iron/L (p=ns); transferrin saturation: M (n=21) 22.10 +/- 13.07, V (n=96) 25.81 +/- 11.79% (p=ns); blood Hb: M (n=22) 107.55 +/-19.70, V (n=98) 111.02 +/- 14.68 g/L (p=ns); MCV: M (n=22) 94.58 +/- 7.35, V (n=98) 93.27 +/- 8.16 fL (p=ns); RDW: M (n=22) 16.60 +/- 1.51, V (n=98) 15.83 +/- 1.39 (p<0.022); soluble transferrin receptors: M (n=18) 1.85 +/- 0.90, V (n=90) 1.89 +/- 0.75 mg/L (p=ns); reticulocyte count: M (n=22) 91.38 +/- 34.69, V (n=98) 87.27 +/- 29.56 *10 9 /L (p=ns); CHR: M (n=22) 31.36 +/- 2.92, V (n=98) 31.46 +/- 3.08 pg (p=ns); IRF: M (n=22) 24.81 +/- 7.55, V (n=98) 23.65 +/- 8.64 (p=ns); intravenous Fe+++ weekly supplementation: M (n=22) 45.45 +/- 26.81, V (n=98) 37.31 +/- 32.25 mg/week (p=ns); a-EPO weekly supple-mentation: M (n=22) 9090.91 +/- 7824.92, V (n=97) 9030.93 +/- 8292.13 UI/week (p=ns). Since it was not feasible to compare an individual event such as death with the spectra of laboratory data or the administered drug amount, the values of each series were in descending order and the number of M patients (approximately 20% of the total) falling into the upper or lower 20% of values was calculated. In front to the expected amount of 4.4 M for each 20%, two M patients fell into the upper 20% and six patients into the lower 20% of blood ferritin values, and, correspondingly, two vs. nine patients for blood iron, four vs. seven patients for transferrin saturation, three vs. seven patients for blood Hb, eight vs. two patients for RDW and six vs. two patients for iron supplementation. Therefore, our patients with a negative prognosis showed an increase in RDW and an iron availability or metabolism disorder. PMID- 15786389 TI - [Quality in dialysis]. AB - BACKGROUND: Several clinical and laboratory dialysis parameters have been suggested by national and international guidelines. Our study aimed to evaluate a series of quality indicators in the dialytic treatment of a group of patients in our two dialytic centers. PATIENTS AND METHODS: In a population of 159 patients on renal replacement therapy (RDT) we performed a 1-yr prospective observational study, with common parameters to indicate cardiovascular pathologies (PAS, LVMI), calcium-phosphorous metabolism (Ca, PO 4 , Ca x PO 4 ), dialytic adequacy (Kt/V, URR%), nutritional status (nPCR, Alb, K), and anemia (hemoglobin (Hb)). For each parameter we evaluated the mean average with the standard deviation and the score percentages below and above the acceptable range (AR). RESULTS: The results demonstrated a substantial stability, with a gradual improvement, in dialytic adequacy, in anemic status and in PAS. The LVMI, calculated at the beginning and at the end of the period, did not demonstrate any significant changes. No significant changes were observed in the calcium-phosphorous metabolism parameters. Serum albumin and nPCR demonstrated the presence of malnutrition in 15-20% of patients. CONCLUSIONS: There was a more than adequate correlation between the target guidelines and that concerning the dialytic adequacy and the corrected anemic status. The improvement in parameters related to cardiovascular pathology, the risk of ectopic calcifications and malnutrition status, represented an achievable target. PMID- 15786390 TI - [Evaluation of central venous catheters by endocavitary ECG]. AB - Jugular vein catheterization (JVC) is adopted for blood access in patients with acute renal failure, chronic renal failure and when patients demonstrate traditional vascular access (VA) failure. We believe that the best technique, first described by Serafini et al, to establish the position of a central venous catheter (CVC) is endocavitary electrocardiography (EC-ECG) and its use is recommended in all uremic patients requiring hemodialysis (HD). This technique uses the tip of the CVC as a reference point in standard electrocardiography. From 2001 to March 2004, we successfully applied this CVC technique in 33 patients requiring HD. The EC-ECG technique is a method that complies with the Food and Drug Administration guidelines regarding catheter tip location in uremic patients. PMID- 15786392 TI - [Type 2 diabetic nephropathy: clinical course and prevention proposals 2004]. AB - The first clinical evidence of nephropathy is the appearance of low, but abnormal, albumin levels in the urine (>30 mg/day or 20 mg/min), microalbuminuria. Without specific interventions, approximately 80% of type 1 diabetics have their urinary albumin excretion increase at a rate of 10-20%/yr to the stage of overt nephropathy or clinical albuminuria (>300 mg/24h or >200 mg/min) over 10-15 yrs, developing hypertension along the way. Approximately 30% of individuals with type 2 diabetes are found to have microalbuminuria or overt nephropathy shortly after the diagnosis of their illness, because diabetes is actually present for many years previously and because the presence of albuminuria can depend on other concomitant nephropathies, as shown by biopsy studies. Without specific intervention, 20-40% of type 2 diabetic patients with microalbuminuria progress to overt nephropathy, but 20 yrs after onset only 20% progress to end-stage renal failure (ESRD). The rates of decline in glomerular filtration rate (GFR) are highly variable from one individual to another, but they may not be substantially different between patients with type 1 and type 2 diabetes. As therapies and interventions for coronary artery disease continue to improve, more elderly type 2 diabetes patients can be expected to survive long enough to develop renal failure. The recently published Italian Society of Nephrology (SIN) guidelines for diagnosis and therapy of diabetic nephropathy present the route for the best strategies in prevention and therapy, from earlier onset to advanced ESRD. PMID- 15786391 TI - [Comparison of methodologies to reach 'dry weight']. AB - BACKGROUND: In clinical practice it is very difficult to determine a final weight that is the expression of normovolemia. In hemodialysis (HD), 'dry weight' is conventionally defined as the weight reached by the patient at the end of that hemodialytic session when the maximum quantity of fluids is removed without inducing any symptomatology. The determination of dry weight has been based on the application of clinical criteria. The use of artificial kidneys with blood volume (BV) sensors has allowed the determination of dry weight through the interpretation of changes in the intradialytic BV curve. Conventional bioimpedance analysis (BIA), or better, the vectorial BIA (BIVA) is a new method for determining dry weight. This study evaluated the use of the above-mentioned method for the proper governing of dry weight. PATIENTS AND METHODS: Twenty HD patients were observed for 4 weeks. In the 1st week, the clinical symptomatology of every patient was monitored during both HD sessions and interdialytic periods. During the 2nd week, intradialytic changes in the BV of each patient were observed on artificial kidneys. In the 3rd week, a cardiologist monitored patients before and after hemodialytic treatments. In the 4th week, the body composition of each patient was analyzed through bioelectrical bioimpedance. RESULTS: Patients, who had clinically shown symptoms of hyperhydration, to the contrary at BIA were dehydrated. Conversely, patients who had dehydration symptoms presented signs of hyperhydration at BIA. CONCLUSIONS: BIVA is the diagnostic instrument that more accurately demonstrates the hydration state of hemodialytic patients. It contributes in defining dry weight more efficiently. PMID- 15786393 TI - [Pregnancy after kidney transplantation. Case load of the Transplantation Center of Vicenza]. AB - BACKGROUND: Pregnancy after kidney transplant has become possible thanks to recent surgical and pharmacological breakthroughs. MATERIALS AND METHODS: We performed a retrospective study including all pregnant women transplanted in our center after 1997. The following variables were analyzed. The type of nephropathy, patient age when dialysis began, patient age at trans-plantation, the time between dialysis and transplantation and the time between transplantation and childbirth. Immunosuppressive therapy, type of delivery, baby's weight and Apgar score were also considered. RESULTS: We followed four pregnancies in three patients who were, respectively, diagnosed with chronic pyelonephritis, post-partum cortical necrosis and immunoglobulin A (IgA) glomerulonephritis (GN). We observed complications in three cases and two pre term births. In one case, the baby's weight at birth was lower when compared to the gestation age. We did not observe any significant disease in the baby's follow-up. CONCLUSIONS: We concluded that our data were in agreement with those in the literature confirming that pregnancy after kidney transplant, although possible, carries an elevated risk; and therefore, patients have to be referred to highly specialized centers. PMID- 15786394 TI - [Molecular Adsorbent Recirculating System in liver function replacement therapy]. AB - The molecular adsorbent recirculating system (MARS) method removes from the blood catabolites either free in the plasma water such as uremic toxins and ammonia, taking advantage of dialysis or free albumin bound ones, like hepatic toxins, transferring them from the albumin in the blood to the albumin circulating in a closed loop where toxins are removed by adsorbtion on resins (charcoal and ion exchange resin). The efficacy of the method in removing the hepatic toxins either in the acute or in the acute on chronic liver failure is demonstrated in numerous studies. Based on these findings, 10 patients affected by acute on chronic liver failure were treated. The results demonstrated that the method, powerfully removing ammonia, bilirubin and bile acids (taken as method efficacy markers), reduced the blood concentrations of these molecules remarkably; allowing the elimination of the refractory pruritus (due to the lowering of plasma bile acid levels), an almost constant symptom in chronic liver diseases, especially with cholestasis, and improves other parameters (cholinesterase, alkaline phosphatase and prothrombin activity). These results agree with those reported in the literature concerning the efficacy of MARS in the replacement of the liver detoxifying function. PMID- 15786395 TI - [Diabetic foot: from diagnosis to therapy]. AB - Four percent of Italians are affected by diabetes mellitus. There are 120 million diabetics worldwide: it is expected that 15% of them will have wounds on their feet during their lifetime. In industrialized countries diabetes mellitus is the main cause of non-accidental amputation and this risk is approximately 15 times higher in diabetics than in any other population. The vasculopathy and/or the diabetic neuropathy represent the basic pathogenic elements in the development of diabetic foot: 15% of diabetics are affected by arteriopathy after 10 yrs of illness and just <50% of diabetics after 25 yrs; 25% of patients have neuropathy after 10 yrs of illness, >40% have neuropathy after 25 yrs. The infection is the third, and often, concomitant pathogenetic factor in the diabetic ulcera. The diagnostics of the vasculopathic diabetic foot makes use of a careful objective check; doppler ultrasonography and arteriography (with the therapeutic application of the PTCA). In addition to the objective check, EMG and the neuropathy autonomy test are fundamental in the neuropathic diabetic foot. Osteomyelitis represents the most fearful complication in the diabetic foot; it can often be solved only through surgical destructive therapy. Therapy of the diabetic foot must come from careful and synergic team work, in which dietician, vascular surgeon, orthopedist and dermatologist make available their own skill with humility and endless patience. PMID- 15786396 TI - [Features of diabetic angiopathy: impotence]. AB - Erectile dysfunction occurs more frequently in the diabetic population than in the general population. This article analyzes a review of this pathology, its prevalence and incidence in the diabetic population, its multifactor origin, and how these causes interact with each other and the therapy required. PMID- 15786397 TI - [Kidney transplantation in Italy: an evolving scenario]. AB - BACKGROUND: Italian transplantation systems have dramatically improved in the last decade. Kidney transplantations are now strictly monitored and excellent results has been achieved in terms of quality and the ability to reduce waiting times for receiving transplantation. METHODS: The Italian organizational retrieval and transplant system is articulated on four levels: local, regional, inter-regional and national. The Italian Transplant Information System (SIT) was set up in 2000 in accordance with Law 91/99. Patient data on the waiting lists and follow-up of transplanted patients are routinely collected. RESULTS: A total of 4406 kidney transplants have been carried out in the 40 Italian kidney transplant centers in 2001-2003. The survival analysis was conducted for the 2000 2002 in 4222 cases. Overall 1-yr survival was 92.4% for the graft and 97% for the patients. After adjusting for variables independently associated with the outcome at multivariate analysis (for example, the case-mix), patient and graft survival at 1 yr was 98.1% and 93.8%, respectively. No remarkable differences in 1-yr graft survival were observed between the 40 Italian kidney transplant centers. At multivariate analysis, variables independently associated with graft failure were donor age, degree of HLA mismatch and recipient case-mix. Analysis of the waiting list showed approximately 6500 patients waiting for kidney transplantation. The mean waiting time was 3.04 yrs, with a mortality rate of 1.18% per year. CONCLUSIONS: Kidney transplantation activity in Italy has produced excellent results in terms of quality and number of transplants per year. However, the number of patients on the waiting list and the waiting time call for further action to increase the number of available organs. PMID- 15786398 TI - [Epidemiology and demography of uremia: data from the Lombardy Dialysis and Transplantation Registry (RLDT)]. AB - The Lombardy Registry of Dialysis and Transplantation (RLDT) since 1983 has collected data concerning patients affected by end-stage renal disease (ESRD) on renal replacement therapy (RRT) in Lombardy, a region of Northern Italy with 9 million inhabitants. This report illustrates the main features of ESRD patients on RRT: there were 6589 patients undergoing treatment at 31 December 2003, with a prevalence rate of 727 pmp. Patient numbers regularly increased by 4.5%/yr for the last 5 yrs. This phenomenon is probably due to the high incidence rate (172 pmp) of ESRD patients in Lombardy during these years related to a relatively stable mortality rate (15.2%). The increasing incidence is probably correlated to the population's characteristics: higher rates (189-223 pmp) were observed in certain provinces (Cremona, Lodi and Pavia) with a larger elderly population (people >65 yrs = >20%, people <65 yrs = <16%). Of dialysis modalities, 85% of prevalent patients were on hemodialysis (HD), 55% in hospital, and 30% in limited care units. The number of patients treated by peritoneal dialysis (PD) was stable during the last years, but showed a slow percentage decline (15% during 2003) since 1999. However, PD remains the first dialysis modality for 21.4% of new patients, with a wide variability among renal units. Regarding HD, highly efficient techniques (on-line hemodiafiltration (HDF)) represented 19.2%, with a significant increase (1.8%) compared to 2002. During 2003, the number of dialysis units in Lombardy was stable; there was only an increase in facility beds in limited care units in order to treat the increasing numbers of uremic patients. PMID- 15786400 TI - [Therapy strategies in the prevention of chronic allograft nephropathy]. AB - Chronic allograft nephropathy (CAN) is an anatomical and clinical alteration, characterized by proteinuria, hypertension and a progressive decline in kidney function, which begins at variable times (months, years) and can lead to the loss of the transplanted organ. CAN pathogenesis, which remains to be fully clarified, involves both immunological (early acute rejection, hyperimmunization, HLA mismatches between donor and recipient, suboptimal immunosuppression, etc) and non-immunological factors (ischemia/reperfusion injury, reduced nephron mass, age differences between donor and recipient, dialysis time, hypertension, dislipidemia, proteinuria, etc). The possible prevention strategies for CAN consist of procedures aimed at the reduction of some potential risk factors: optimization of the conditions for organ explantation, diminution of ischemia/reperfusion injury, aggressive pharmacological treatment of acute rejection episodes, routine utilization of anti-hypertensive and hypolipidemic agents, and appropriate and rational immunosuppressive regimen. Moreover, some categories of immunosuppressive drugs, such as calcineurin inhibitors, can have a nephrotoxic effect, often regardless of therapeutic dosage. The introduction in clinical practice of novel immunosuppressive drugs with no nephrotoxicity, like mycophenolate mofetil and rapamycin, makes therapeutical strategies able to reduce the incidence of CAN feasible. PMID- 15786399 TI - [Renal transplantation in the North Italy Transplant program (NITp): Organ allocation and results]. AB - Renal transplantation is an effective therapeutic tool for patients with end stage renal diseases (ESRDs). Data reported in this article summarize the results obtained from 30 years' activity in the North Italy Transplant program (NITp), the first transplant organization in Italy that implemented a donor procurement and organ transplantation network. In the NITp kidney allocation is governed by a computerized algorithm, NITK3, put in place in 1997, aimed at ensuring equity, transparency and traceability during the stages of the allocation decision-making process. The NITp working group has recognized the NITK3 criteria and they are periodically reviewed following the results of the analysis of patients' transplantation odds. The results obtained with the use of the NITK3 algorithm have been very satisfactory: after 6 yrs, a significantly higher percentage of patients at immunological risk (sensitized or waiting for re-transplant), of patients waiting for >3 yrs and of patients with 0-1 HLA A,B,DR mismatches have been transplanted. Moreover, a higher percentage of kidneys were used locally (in a hospital within the procurement area), and this is known to stimulate donor procurement. Finally, we performed a preliminary statistical analysis of transplants carried out from 1998-2002 in 5/16 centers of the NITp area, demonstrating the quality of the NITp program in terms of patient and graft survival, and that donor and recipient age are the variables significantly impacting on transplant results. PMID- 15786401 TI - [Anemia treatment to reduce mortality risk and to improve quality of life in chronic uremic patients]. AB - Survival of uremic patients on dialytic treatment is significantly worse compared with that of the general population, mainly because of cardiovascular disease (CVD) excess. Anemia, a frequent and relatively early complication of impaired renal function, can considerably worsen the outcome for these patients. Due to the induced alterations on cardiovascular structures, first, left ventricular hypertrophy, anemia is not only a condition significantly impairing quality of life, but also a serious threat for the long-term survival of patients undergoing dialysis. Several studies actually showed the existence of a clear inverse association between hemoglobin (Hb) levels and mortality or hospitalization rates in patients with renal failure. Although the benefits of a partial correction of Hb levels, even in terms of left ventricular hypertrophy regression, have been well documented, it remains unclear whether starting the treatment in a very early phase of the disease or achieving a complete normalization of Hb levels above the target values recommended by current guidelines can provide further advantages, at least in selected patient groups. However, ongoing clinical trials, particularly the CREATE and the ACORD, will be able to clarify better which anemia correction practices can ensure the best results both for quality of life and for cardiovascular status; and therefore, for the long-term survival of patients with renal disease. PMID- 15786402 TI - [Control of calcium and phosphate metabolism and prevention of vascular calcifications in uremic patients]. AB - Vascular calcifications are more frequent in dialysis patients than in the general population or in patients with cardiovascular disease (CVD) and normal renal function. The reasons for this high incidence are multiple; they include traditional factors such as hypertension, diabetes, dyslipidemia, and specific factors such as sodium overload, hyperomocysteinemia, chronic inflammation and oxidative stress, as well as mineral metabolism disturbances. Specifically, hyperphosphatemia and the elevated calcium (Ca) x phosphate product have been associated with an increased risk for the development of vascular calcification and death. Treatment with Ca salts can induce hypercalcemia, increased Ca x phosphate product and Ca overload. Sevelamer substitution for Ca salts has been documented to attenuate the progression of coronary artery and aortic calcification. A possible mechanism explaining this observation could be ongoing Ca loading related to oral Ca ingestion. Treatment with Ca salts could induce Ca overload, particularly in patients dialyzed against a high dialysate Ca (>1.5 mmol/L) solution, which is known to determine a positive dialysis balance. Conversely, an overall negative Ca balance can result from low Ca dialysate use (1.25 mmol/L) when the patients do not receive Ca supplements or vitamin D metabolites. Maintaining normal Ca and phosphate balances remains a primary goal in the management of dialysis patients. Control of hyperphopshataemia should be achieved either using Ca and aluminum-free phosphate binders, such as sevelamer, or Ca salts, alone or in combination, provided that a daily oral elemental Ca intake of 1.5 g is not exceeded. PMID- 15786404 TI - [Arterial hypertension and cardiovascular risk in patients with nephropathy]. AB - In the general population, there is a very close link between arterial hypertension and cardiovascular diseases (CVDs); hypertension is the most important predictive factor for coronary disease. In the patient with renal insufficiency, along with the classical factors behind vascular damage, there are those linked to uremia such as oxidative stress, hyperhomocysteinemia, ADMA and chronic microinflammation, responsible for the process of early atherosclerosis. Nevertheless, many questions remain unanswered concerning the optimum pressure target, both in terms of which arterial pressure could be more indicative of organ damage and the ideal pressure range for the patient. On the one hand, recent studies in populations with advanced renal insufficiency have cast doubts on the close relationship between hypertension and cardiovascular mortality, attributing a more unfavorable prognostic weight to hypotension than to hypertension. On the other hand, a new concept concerning arterial pressure has been developing over recent years, i.e. differential pressure, which is supposed to represent a vascular damage index, in terms of wall rigidity; and therefore, arterial compliance, which is more reliable, complete and dynamic as compared to the traditional pressure markers such as systolic, diastolic and mean blood pressure (BP). This renders our attempt to control the risk factors as somewhat limited, with the sole control of systolic BP and diastolic BP in absolute terms. Control that to judge from the various studies published to date, appears to be far from easy to implement in clinical practice, and above all in dialysis. PMID- 15786403 TI - [Prevention of extraskeletal calcifications in uremia]. AB - Secondary hyperparathyroidism (HPTH) is a common feature in end-stage renal disease (ESRD) patients. The three main factors involved in secondary HPTH pathogenesis are high phosphate levels, hypocalcemia and vitamin D deficiency. Recently, many studies demonstrated a strong association between bone disease and cardiovascular events in chronic kidney disease patients. In addition, cardiovascular events are the most frequent cause of death in patients with chronic renal failure. Increased levels of serum phosphorus and calcium-phosphate product are directly involved in the pathogenesis of extraskeletal calcifications (blood vessels, soft tissues, etc) in dialyzed patients compared to the non uremic population. Recent studies suggested that vascular calcification is due not only to a passive calcium-phosphate deposition on atherosclerotic arteries, but also to active mechanisms regulated by bone-associated genes. In particular, fetuin and matrix Gla-protein are two 'protective' proteins associated with reduced vascular calcification and could be the regulatory keys in preventing this process in renal failure. The limitations of calcium salts as phosphate binders in patients with advanced renal failure have been thoroughly evaluated in the last 5 yrs. New phosphate binders, which do not contain aluminum or calcium, have been developed to reduce the risk of extraskeletal calcifications in ESRD. PMID- 15786405 TI - [A good vascular access allows an effective treatment]. AB - Vascular access (VA) for dialysis is defined as the 'Achilles heel', but also the 'Cinderella' of dialysis, indicating the poor consideration of the problem whether in the surgical environment, or in incomprehensible way in that nephrologic. It can only aspire to the definition 'Fundamental detail'. However, presupposed effective dialysis is a blood flow rate of 300-350 mL/min. Good VA must be easy to prepare, long lasting, free from complications, and aesthetically acceptable and economical. The arteriovenous fistula (AVF) of Cimino and Brescia, from 1966, represents the gold standard and the model of comparison for other systems, more technologically advanced. It must be programmed with an adapted margin (1-2 months) to allow maturation and access certainty for the first puncture, and never carried out sooner than 14 days from the operation. It is known from hemodynamic studies that the good functional flow of the new fistula can already regain 400-500 ml/min in the first week, with cardiological implications like the increase in cardiac throw, in ejection fraction and in the cardiac index. Health workers, patients and dialysis staff must follow a continuous educational program to protect the VA and avoid 'routine and absent minded management', a basis for its premature failure. The nephrologist must take the responsibility upon himself not to carry out 'medical malpractice'. In the Dialysis Center of Mantova, the VA 'road map' previews all patients (young and old, affections from mono or pluropathology), first the fistula to the wrist, then the cephalic proximal. It follows the basilic vein transposition, the vascular graft to the arm or to the groin, as an alternative to peritoneal dialysis. The permanent central venous catheter (CVCp) is the last choice in patients with reduced life expectancy, heart failure, neoplastic patients with vascular patrimony destroyed by chemotherapy and ischemic lesions produced by the fistula. There were 180 afferent prevailing patients at the Mantova Dialysis Center . The natural fistula rate was 91%, grafts 7% and CVCps 2%. Between 2000 and 31 March 2004 we prepared 367 VAs. Average patient age was 65 yrs, range 20 90 yrs; 59% male and 41% female. Eighty-eight percent of operations were performed by the nephrologist (distal fistula, rescue and cephalic proximal) and 12% by the vascular surgeon (basilic vein transposition, graft in PTFE stretch to the arm and to the groin and permanent catheter in the jugular vein). The fistula with native veins was the better solution, the graft must be prepared after the exhaustion of natural possibilities, and the CVCp, for serious complications (inadequate flow and infections), must be the last alternative. PMID- 15786406 TI - [Water quality cuts down the chronic inflammatory risk: how to obtain it and keep it over time]. AB - BACKGROUND: Every week, approximately 400 liters of water used for dialysate production come into direct contact, through the semi-permeable membrane of the dialyzer, with the dialysis patient's blood stream. Therefore, submitting municipal water to an adequate depuration process before its use for dialysis becomes necessary. METHODS: Problems related to the implementation, updating and management of a dialysis water treatment system are analyzed. The results of the most recent multicenter studies on dialysis fluids quality are also reviewed. RESULTS: The best approach to plan, implement and manage a dialysis water treatment system, first, consists of defining the standards of chemical and microbiological water quality. The most diffused and commonly accepted standards are those recommended by the Association for Advancement of Medical Instrumentation (AAMI) and the European Pharmacopea (EP), which allow a maximum bacterial growth of, respectively, 200 CFU/ml and 100 CFU/mL and a maximum endotoxin concentration of 2 IU/mL and 0.25 IU/mL. A modern dialysis water treatment system provides a final purification process, mainly by reverse osmosis (RO), together with different pre-treatment levels and a hydraulic distribution circuit. Therefore, as RO produces water of optimal chemical and microbial quality, all efforts in the dialysis unit must be aimed at keeping this quality as constant as possible over time, by carrying out effective maintenance strategies and system disinfection. Nevertheless, several multicenter studies reported that 7-35% of water samples exceed a bacterial growth of 200 CFU/mL and that 44% of them display endotoxin concentrations >5 IU/mL. CONCLUSIONS: The results of multicenter studies indicate that the microbial quality of dialysis fluids is, unfortunately, still an often neglected problem. Evidence of a possible relationship between dialysis fluid contamination and patient morbidity, as well as the availability of systems and machines allowing purity levels that were unimaginable only a few years ago, must be a stimulus for modifying clinical practices and starting the improvement processes aimed at maximally reducing the risk of microbial contamination in the dialysis water, as already done with chemical contamination. PMID- 15786407 TI - [Coronary disease: early intervention saves lives. Treatment of acute myocardial infarction in Mantova: results of thirty months of work after the implementation of a province network]. AB - BACKGROUND: Since June 2001, in the province of Mantova, we have undertaken a program for the management of acute myocardial infarction based on the early assessment of patient risk profiles, concerning telematic connections among care centers and on the optimization of in-hospital and out of hospital critical pathways for access to care. MATERIALS AND METHODS: Our network provides connections among the following centers: advanced life support ambulances, seven hospitals, three coronary care units, one cath lab on call 24 h a day for primary angioplasty, and one thoracic surgery division. This program, through its strong telematic platform, allows the early assessment of myocardial infarction and provides primary angioplasty to all high-risk patients, as fibrinolytic treatment is reserved only for low-risk patients admitted in peripheral hospitals. RESULTS: Two hundred and eighty patients with acute myocardial infarction were treated with angioplasty; 224 patients (80%) underwent primary angioplasty, 36 patients (13%) facilitated angioplasty and 20 patients (7%) rescue angioplasty. One hundred and thirty-two patients (47%) were first admitted to Mantova Hospital; 78 patients (28%) were referred to Mantova from peripheral hospitals and 70 patients (25%) were directly transported to the cath lab by advanced life support ambulances. Procedural success was obtained in 98% of patients, with 0.4% intraprocedural mortality. In-hospital mortality was 5.7%, while mortality in cardiogenic shock patients was 36%. The recurrence of acute myocardial infarction occurred in 1% of patients and major bleeding occurred in 2.2% of patients. One patient with cardiogenic shock died during transport. Mean door to balloon time was 67 min with a 42% reduction in the 3rd recruitment period. CONCLUSIONS: This program, developed in the setting of a provincial network for the management of acute myocardial infarction, provided primary angioplasty to all high-risk patients, with a high procedural success rate. Within a few months, time to treatment was minimized by the use of telematic facilities. PMID- 15786408 TI - [Peripheral arteriopathy in ESRD dialysed patients: when and how to intervene]. AB - Critical limb ischemia secondary to chronic peripheral occlusive disease is common in chronically dialysed patients, with an incidence rate of 25-30%. Atherosclerotic lesions are more frequent in the infrainguinal arteries and long infrapopliteal occlusions often occur. Due to diabetes, hypertension and ischemic cardiopathy, the surgical prognosis is very poor in these patients; medical treatment should always be attempted associated with analgesia, without an excessive delay in surgical therapy if needed. Both spinal stimulation and lumbar simpaticectomy often fail; open and endovascular surgery are the best options before major amputation, which has a high incidence in this patient subgroup. Between 2000 and 2003, 23 chronically dialysed patients underwent surgery. Nine open and 13 endovascular procedures were performed, associated with four immediate and five late minor amputations. Despite an immediate mortality rate of 8.6%, we obtained immediate patency and limb salvage in all cases. In a medium follow-up of 25 months (range 3-36), five thromboses were found in subinguinal procedures; not one in iliac procedures. The five patients underwent major amputation. Another two patients underwent amputation despite arterious patency. Seven patients died due to cardiovascular diseases during the follow-up. Our experience confirms that the association between POAD and dialysis is a prediction factor for medium-term death and that the surgical risk is highly increased. It is important to select patients undergoing surgical treatment to check for the lowest invasivity. PMID- 15786409 TI - [Hypertension, atherosclerosis and kidney]. AB - Cardiovascular risk is dramatically increased in patients with end-stage renal disease (ESRD). However, even minor dys-functions such as microalbuminuria or a mild increase in serum creatinine (Cr) have a major impact on cardiovascular risk. Increased cardiovascular risk is present in multiple populations, including general populations, patients with moderate risk such as hypertensives, and high risk patients including patients with heart failure and myocardial necrosis. There are many mechanisms underpinning the increased cardiovascular risk. Regarding atherosclerosis, the kidney can be victim or villain. On the one hand, both kidney disease per se and renal insufficiency can induce vascular damage, thereby increasing cardiovascular risk. Kidney disease without renal insufficiency can cause an increased prevalence in hypertension, dyslipidemia (nephrotic syndrome), sympathetic system hyperactivity, and in renin angiotensin system hyperactivity. A moderate-severe renal insufficiency can induce an increase in many vasculotoxic substances such as ADMA, lipoprotein(a), homocysteine, disturbances in calcium and phosphate metabolism, anemia and left ventricular hypertrophy. A more severe renal insufficiency can induce the ominous malnutrition-inflammation-atherosclerosis (MIA) syndrome. On the other hand, the kidney can be the victim of atherosclerosis. Ischemic nephropathy, caused by atherosclerotic renal artery disease and atheroembolism from abdominal aorta are two examples. Finally, it is important to consider that the kidney, being an organ with a wide vasculature, could be a sophisticated sensor of subclinical cardiovascular damage. PMID- 15786410 TI - [Infection risk and personal protective devices]. AB - The Aa discuss the evidence and recommendations for the use of personal protective equipment by health care practitioners in general care settings; the use of aprons, gowns, gloves, eye protections, face masks is valutated on the basis of an assessment of the risk of transmission of microrganisms to the patient or from the patient to health care practitioners. The primary uses of personal protective equipment are to protect staff and reduce opportunities for transmission of microrganisms in hospitals. A trend to eliminate the unnecessary wearing of aprons, gowns and masks in general care settings has evolved over the past twenty years due to the absence of evidence that they are effective. PMID- 15786411 TI - [Nurses' professional responsibility]. AB - As managed care relevance is growing, several old issues related to personal institutional responsibility are increasing among practitioners. Therefore, as a professional figure a nurse bases his/her job on a mix of personal knowledge and skills along with training, and he/she is responsible for giving advice in line with professional care standards. In addition, he/she is in charge of the treatment pattern agreed with the patient. However, nursing is a much more complex job, which leads professional figures facing the controversial issue of combining institutional responsibility and nursing professional tasks and duties daily. As far as nursing institutional responsibility is concerned, different view points or approaches can be applied to investigate it. The most common one is the legal approach, yet this is not the most appropriate one. Therefore, our professional background is mainly based on a management prospective rather than a legal one; dealing with the issue legally would lead, essentially, to a summary of laws and regulations without any kind of argumentative discussion. Consequently, this study aimed to analyze nurses' institutional responsibility by approaching the issue from an innovative human resources management prospective; therefore, defining the gap between nursing institutional responsibility and its tasks. PMID- 15786412 TI - Prognostic significance of circumferential margin involvement in rectal adenocarcinoma treated with preoperative chemoradiotherapy and low anterior resection. AB - INTRODUCTION: Histologic examination of circumferential margins is an important predictor of local and distant relapse in non-radiated rectal cancer. However, for patients who received preoperative chemoradiotherapy this role has not yet been addressed. METHODS: From January 1995 to December 1997, 61 patients with rectal adenocarcinoma located between 0 and 10 cm from anal verge with invasion into perirectal fat assessed by rectal ultrasound were included. All patients received 45 Gy + bolus infusion of 5-FU (450 mg/m(2)/days 1-5, 28-33 of RT); 4-6 weeks later, surgery was performed. Circumferential margin was assessed (<2 mm was considered as positive). Five-year survival was calculated by Kaplan-Meier method and comparison of groups with log-rank test. Multivariate Cox regression analysis was performed to find risk factors affecting local control and survival. RESULTS: There were 35 males and 26 females, mean age 60.3 years. Twelve patients (19.7%) had circumferential margin involvement. Median follow-up was 44 months. Overall local recurrence was observed in 6 of 61 patients (9.8%); in patients without circumferential margin involvement this was 8%, whereas it was 16% in those with circumferential margin involvement (P = 0.33). Distant recurrence was observed in 22% of patients without circumferential margin involvement; conversely, it was 58.3% in those with involvement (P = 0.02). Five-year survival of patients without circumferential resection involvement margin was 81%, while it was 42% in patients with circumferential involvement (P = 0.006). CONCLUSIONS: In patients with rectal cancer treated by preoperative chemoradiation plus total mesorectal excision (TME) and sphincter saving surgery, circumferential margin involvement is associated with high incidence of distant recurrence and cancer related death. PMID- 15786413 TI - Spontaneous rupture of adrenal pheochromocytoma: review and analysis of prognostic factors. AB - BACKGROUND AND OBJECTIVES: Because of its rarity, the clinical characteristics of the manifestation of a ruptured pheochromocytoma and factors influencing on treatment outcomes in patients are still unclear. METHODS: A comprehensive MEDLINE review of the literature on pheochromocytomas, rupture and hemorrhage was performed. Based on the review and one additional case of our hospital, statistical analysis was undertaken to analyze the association of clinical outcome with clinical presentation, preoperative diagnosis, and treatment modality. RESULTS: We identified 49 other cases of hemorrhagic pheochromocytoma in the literature. After our case was added, a review of all 50 cases revealed that 17 of 38 that could not undergo adequately prepared (elective) surgery died, while all 12 patients who underwent elective surgery survived (P = 0.004). An accurate preoperative diagnosis of pheochromocytoma was associated with elective surgery (P < 0.0001), and hemodynamic instability (shock status) was a significant factor for a failed diagnosis and inadequate management, resulting in a poor prognosis (P = 0.023). CONCLUSIONS: Although it is a very rare condition, physicians should be aware that a pheochromocytoma can bleed and present acutely in the abdomen with shock; an accurate diagnosis and adequately prepared surgical removal are important for a good postoperative prognosis. PMID- 15786414 TI - How I do it: bi-segmentectomy V-VIII as alternative to right hepatectomy: an intrahepatic approach. PMID- 15786415 TI - Phenotypic variability in myotonia congenita. AB - Myotonia congenita is a hereditary chloride channel disorder characterized by delayed relaxation of skeletal muscle (myotonia). It is caused by mutations in the skeletal muscle chloride channel gene CLCN1 on chromosome 7. The phenotypic spectrum of myotonia congenita ranges from mild myotonia disclosed only by clinical examination to severe and disabling myotonia with transient weakness and myopathy. The most severe phenotypes are seen in patients with two mutated alleles. Heterozygotes are often asymptomatic but for some mutations heterozygosity is sufficient to cause pronounced myotonia, although without weakness and myopathy. Thus, the phenotype depends on the mutation type to some extent, but this does not explain the fact that severity varies greatly between heterozygous family members and may even vary with time in the individual patient. In this review, existing knowledge about phenotypic variability is summarized, and the possible contributing factors are discussed. PMID- 15786416 TI - Nutritional inadequacy in adults with muscular dystrophy. AB - Patients with muscular dystrophy may be prone to nutrient deficiency due to mobility limitations or oropharyngeal weakness. Patients with myotonic muscular dystrophy (DM1) may be particularly prone to nutritional deficiencies from associated dysmotility of the entire gastrointestinal tract. We prospectively evaluated nutritional intake, body composition, and muscle strength in adult patients with DM1 (n = 29) and other muscular dystrophies (n = 22) on two occasions separated by approximately 6 months. Handgrip was significantly lower and knee extension higher for DM1 compared to other dystrophies, with no between group differences in nutritional intakes. Many patients in both groups demonstrated inadequate nutrient intake of protein, energy, vitamins (water and fat soluble), and minerals (calcium and magnesium). Significant correlations were found between measures of strength and certain individual nutrients (e.g., copper and water-soluble vitamins). These data indicate that a substantial number of adults with muscular dystrophy do not meet current dietary intake recommendations. The potential clinical implications of these findings are discussed. PMID- 15786417 TI - Excitatory synaptic potentials in spastic human motoneurons have a short rise time. AB - This study assessed whether changes in size or time-course of excitatory postsynaptic potentials (EPSPs) in motoneurons innervating spastic muscle could induce a greater synaptic response, and thereby contribute to reflex hyperexcitability. We compared motor unit (MU) firing patterns elicited by tendon taps applied to both spastic and contralateral (nonspastic) biceps brachii muscle in hemiparetic stroke subjects. Based on recordings of 115 MUs, significantly shortened EPSP rise times were present on the spastic side, but with no significant differences in estimated EPSP amplitude. These changes may contribute to hyperexcitable reflex responses at short latency, but the EPSP amplitude changes appear insufficient to account for global differences in reflex excitability. PMID- 15786418 TI - Successful human ovarian autotransplantation to the upper arm. PMID- 15786420 TI - Comparison of HER-2 status determined by fluorescence in situ hybridization in primary and metastatic breast carcinoma. AB - BACKGROUND: Accurate assessment of HER-2 status is necessary prior to anti-HER-2 antibody (trastuzumab) therapy for metastatic breast carcinoma. However, controversy exists regarding whether to assess HER-2 status in the primary tumor or in metastatic lesions. It is also unclear whether HER-2 status can change during disease progression or after chemotherapy. METHODS: Breast carcinoma samples from 60 women with known HER-2 status in both primary tumors and paired metastases (locoregional disease, n = 43 patients; distant disease, n = 17 patients) were reviewed retrospectively. Thirty-two patients underwent chemotherapy before their metastatic lesions were sampled, including 18 patients who received neoadjuvant chemotherapy and 14 patients who received adjuvant chemotherapy. The HER-2 gene was examined by fluorescence in situ hybridization either in paraffin-embedded tissue samples (48 primary tumors and 9 metastatic tumors) or in fine-needle aspirates (12 primary tumors and 51 metastatic tumors). HER-2 gene amplification was defined as a HER-2:chromosome 17 signal ratio >/= 2.0. RESULTS: The HER-2 status of primary and metastatic tumors agreed in 58 of 60 patients (97%), including 18 (30%) amplified tumors and 40 (67%) nonamplified tumors. A discrepancy in HER-2 status was observed in specimens from two patients in which HER-2 amplification was detected in the primary tumor but not the metastatic tumors. In one patient, three foci of tumor nodules were found in the same breast; the HER-2 status was assessed in only one of them, which showed amplification; however, HER-2 amplification was not detected in the axillary lymph node metastasis. In another patient, the HER-2 gene was amplified in the primary tumor but not in the liver metastasis. No metastases showed HER-2 amplification without amplification in the primary tumor. Locoregional and distant metastases demonstrated similar concordance rates with their corresponding primary tumors (98% and 94%, respectively). Complete concordance of HER-2 status was found between primary tumors prior to chemotherapy and metastases that were sampled after chemotherapy. CONCLUSIONS: The HER-2 status in breast carcinoma generally was stable during metastasis, whether to locoregional or distant sites. Chemotherapy did not modify the HER-2 status in metastatic lesions. Therefore, HER-2 amplification can be evaluated reliably in material from either primary or metastatic tumors in most patients. Further study with larger series is warranted to elucidate the significance of discordant results. PMID- 15786422 TI - Phase I/II trial of adding semisynthetic homoharringtonine in chronic myeloid leukemia patients who have achieved partial or complete cytogenetic response on imatinib. AB - BACKGROUND: A Phase I/II study was designed to show whether the addition of semisynthetic homoharringtonine (sHHT) would reduce the level of residual disease in patients with Ph-positive chronic myeloid leukemia who appeared to have achieved a suboptimal response to imatinib alone. METHODS: Patients with CML who had achieved >/= 35% Ph-negativity on imatinib were included. All patients had been treated with imatinib at >/= 400 mg/day for at least 2 years and had achieved a plateau in BCR-ABL transcripts defined by measuring BCR-ABL transcripts on at least 4 occasions over a minimum period of 1 year with the latest value not lower than the previous minimum value. Initially sHHT was given subcutaneously at a dose of 1.25 mg/m(2) twice daily for 1 day. Courses were repeated every 28 days. The dosage of sHHT was escalated by adding one day of treatment every two days. Efficacy was assessed by serial monitoring of blood levels of BCR-ABL transcripts. RESULTS: Of 10 evaluable patients, 7 had an appreciable decline in BCR-ABL transcript levels; in 5 cases the reduction was greater than 1 log. Asthenia (n = 10) and cytopenias (n = 3) were prominent side effects, but the drug was generally well tolerated. Mutations in the P-loop of the BCR-ABL kinase domain were found in 2 of the patients who responded to the addition of sHHT. CONCLUSIONS: The addition of sHHT should be considered for patients on imatinib who fail to obtain low levels of minimal residual disease. PMID- 15786421 TI - Chemosensitization by STI571 targeting the platelet-derived growth factor/platelet-derived growth factor receptor-signaling pathway in the tumor progression and angiogenesis of gastric carcinoma. AB - BACKGROUND: Autocrine and paracrine growth mediated by the platelet-derived growth factor (PDGF)/PDGF receptor (PDGFR)-signaling pathway plays an important role in the progression of solid tumors. The authors assessed the effect of STI571 on the tumor growth of gastric carcinoma in combination with 5 fluorouracil (5-FU) or paclitaxel targeting the PDGF/PDGFR-signaling pathway. METHODS: In MKN-45 gastric carcinoma cells, the cytotoxic effect was evaluated by 3-(4,5 dimethiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, and the in vivo antitumor effect was evaluated in a nude mouse xenograft. Both STI571 and an antitumor drug were administered intraperitoneally. Gene expression was assessed by Western blot analysis and immunohistochemical staining. Apoptotic cell death was evaluated by the terminal deoxyuridine triphosphate-biotin nick-end labeling assay, and tumor angiogenesis was evaluated by microvessel density analysis. RESULTS: Treatment with STI571 alone was not effective in vitro, as assessed by a 50% inhibitory concentration value of 24.3 microM. Combination treatment with STI571 and 5-FU or paclitaxel enhanced the cytotoxic effect somewhat when the concentration of STI571 was increased to 10 microM. Combination treatment with STI571 and 5-FU or paclitaxel enhanced the antitumor effect of the antitumor drug significantly in vivo. The enhanced antitumor effect was associated with increased apoptotic cell death and inhibition of tumor angiogenesis. Treatment with STI571 down-regulated the expression of PDGF-BB and PDGFR-beta in tumor cells and decreased the production of phosphorylated PDGFR-beta and phosphorylated Akt. Furthermore, treatment with STI571 inhibited the expression of PDGFR-beta in stromal cells. CONCLUSIONS: STI571 was an effective chemosensitizer of antitumor drugs, such as 5-FU and paclitaxel for gastric carcinoma, targeting the PDGF/PDGFR-signaling pathway of tumor cells and stromal cells in disease progression and angiogenesis. PMID- 15786424 TI - Provider volume and outcomes for oncological procedures. AB - BACKGROUND: Oncological procedures may have better outcomes if performed by high volume providers. METHODS: A review of the English language literature incorporating searches of the Medline, Embase and Cochrane collaboration databases was performed. Studies were included if they involved a patient cohort from 1984 onwards, were community or population based, and assessed health outcome as a dependent variable and volume as an independent variable. The studies were also scored quantifiably to assess generalizability with respect to any observed volume-outcome relationship and analysed according to organ system; numbers needed to treat were estimated where possible. RESULTS: Sixty-eight relevant studies were identified and a total of 41 were included, of which 13 were based on clinical data. All showed either an inverse relationship, of variable magnitude, between provider volume and mortality, or no volume-outcome effect. All but two clinical reports revealed a statistically significant positive relationship between volume and outcome; none demonstrated the opposite. CONCLUSION: High-volume providers have a significantly better outcome for complex cancer surgery, specifically for pancreatectomy, oesphagectomy, gastrectomy and rectal resection. PMID- 15786423 TI - Residual insulin secretion at diagnosis of type 1 diabetes is independently associated with both, age of onset and HLA genotype. AB - BACKGROUND: We investigated whether residual insulin secretion and metabolic derangement at diagnosis of type 1 diabetes (T1DM) are influenced by human leukocyte antigens (HLA) class II genes. METHODS: Eight hundred and seventy-one T1DM consecutive Caucasian patients were typed for HLA class II genes. In 300 of these patients, glycated haemoglobin, insulin requirement, baseline C-peptide and body mass index (BMI) Z-score were measured at clinical diagnosis. The effect of the HLA genotypes on the quantitative variables was investigated using multiple linear regression. The beta coefficient regression of the age at onset and HLA genotypes were standardized to compare their specific importance for C-peptide levels. RESULTS: The HLA genotypes were divided in high-, moderate- and low-risk categories. The frequency of high-risk genotype, DRB1*03-DQB1*0201/DRB1*04 DQB1*0302, decreased with increasing age of onset (p < 0.0001, chi(2) linear trend). The presence of the high-risk genotype was independently associated with lower C-peptide levels at diagnosis (p = 0.002). In the regression analysis of C peptide levels, the standardized beta coefficient for age of onset and high risk compared to low-risk genotypes showed similar results (0.27 and 0.24 respectively). There was a positive association between age of onset and C peptide (p < 0.0001) and a negative association between age of onset and insulin requirement (p < 0.0001). CONCLUSIONS: The degree of beta-cell destruction at diagnosis of T1DM is independently associated with both, age of onset and HLA genotypes, the two variables exert a similar quantitative effect on residual beta cell function at diagnosis. PMID- 15786425 TI - Body parts from the laboratory bench. PMID- 15786426 TI - Microarray technology for surgeons. PMID- 15786427 TI - Randomized clinical trial of mechanical bowel preparation versus no preparation before elective left-sided colorectal surgery. AB - BACKGROUND: Mechanical bowel preparation (MBP) is performed routinely before colorectal surgery to reduce the risk of postoperative infectious complications. The aim of this randomized clinical trial was to compare the outcome of patients who underwent elective left-sided colorectal surgery with or without MBP. METHODS: Patients scheduled for elective left-sided colorectal resection with primary anastomosis were randomized to preoperative MBP (3 litres of polyethylene glycol) (group 1) or surgery without MBP (group 2). Postoperative abdominal infectious complications and extra-abdominal morbidity were recorded prospectively. RESULTS: One hundred and fifty-three patients were included in the study, 78 in group 1 and 75 in group 2. Demographic, clinical and treatment characteristics did not differ significantly between the two groups. The overall rate of abdominal infectious complications (anastomotic leak, intra-abdominal abscess, peritonitis and wound infection) was 22 per cent in group 1 and 8 per cent in group 2 (P = 0.028). Anastomotic leak occurred in five patients (6 per cent) in group 1 and one (1 per cent) in group 2 (P = 0.210) [corrected] Extra abdominal morbidity rates were 24 and 11 per cent respectively (P = 0.034). Hospital stay was longer for patients who had MBP (mean(s.d.) 14.9(13.1) versus 9.9(3.8) days; P = 0.024). CONCLUSION: Elective left-sided colorectal surgery without MBP is safe and is associated with reduced postoperative morbidity. PMID- 15786428 TI - New parallel software (P_Anhar) for anharmonic vibrational calculations: application to (CH(3)Li)(2). AB - We present the development of a new parallel computer code (P_Anhar_v1.0) to calculate the vibrational spectrum of medium size molecules using a variational algorithm. The method is applied to the determination of a complete quartic anharmonic force field (B3LYP/cc-pVTZ) for methyllithium, leading to a new interpretation of experimental data. PMID- 15786429 TI - Exploring the potential energy surface of retinal, a comparison of the performance of different methods. AB - The ground state structure of retinal has been investigated. We found that DFT and CASSCF produce different results for the bond length alternation in a model system of retinal. Quantum mechanics/molecular mechanics calculations including the closest surrounding amino acids have been performed, using DFT and CASSCF to calculate the structure of retinal in the protein cavity. The planarity of the retinal molecule is affected by the surrounding protein. DFT and CASSCF produce different twist angles. The difference between CASSCF and DFT appears to be related to the positively charged nitrogen of the Schiff base, which leads to different pi-bond orders produced by the two methods. PMID- 15786430 TI - Cryoablation and cryolocalization in the management of breast disease. AB - Cryotechnology is currently used for both treatment and diagnosis of breast disease. Due to the natural analgesic effect of cold, cryoablation is potentially more patient-friendly than other technologies which raise tissue temperature. Freezing produces a predictable volume of necrosis and is easily observed and controlled during treatment. Recent studies have demonstrated that, as a primary therapy for breast fibroadenoma, cryoablation is safe and effective with durable results that can be reproduced in community practices. Certain barriers do exist before cryoablation, or any other in situ ablation, can become a standard therapy for the treatment of localized breast malignancy. Investigations are underway to refine patient selection criteria and develop valid confirmatory assays so that clinical trials can begin. Cryolocalization, which creates a well-delineated, palpable mass of frozen tissue encompassing a tumor, is a relatively new application of cold in medicine. This strategy promises to reduce positive margin rates during lumpectomy of non- or barely-palpable tumors. Finally, cryotechnology now also aids in the collection of tissue for histological analysis. PMID- 15786431 TI - Site-select procedure for non-palpable breast lesions: feasibility study with a 15-mm cannula. AB - BACKGROUND: As screening mammography becomes more commonplace, increasing numbers of non-palpable breast lesions are being found. The aim of this prospective study was to evaluate the feasibility, utility, and patient-perceived cosmesis and satisfaction of the Site-Select procedure in women with non-palpable breast lesions. METHODS: Thirty-two consecutive patients underwent the Site-Select procedure, performed under local anesthesia by the same surgeon. The Site-Select procedure was included in a protocol for small (<15 mm) breast imaging reporting and data system (BI-RADS) grade 3 and 4 breast lesions. The pathologic diagnosis, specimen size, length of the procedure, perioperative and postoperative complications, subsequent interventions, patient satisfaction, and esthetic results were documented. RESULTS: The Site-Select procedure was successful in all 32 patients (mean age, 56 years; range, 44-79 years). Mammographic lesions corresponded to microcalcifications in 21 patients, microcalcifications with architectural distortion in 4 patients, microcalcifications with nodules in 2 patients, and architectural distortion alone in 5 patients. The Site-Select procedure was used on an outpatient basis. Carcinomas were diagnosed in five patients (15.6%). No complications occurred during the procedure. The only postoperative complication was a hematoma, which did not require surgical drainage. No missed cancers were detected by follow-up mammography (mean 8 months later; range, 1-18 months). The esthetic results and patient satisfaction were excellent. CONCLUSIONS: This study demonstrates that the Site-Select procedure is an effective diagnostic method in selected cases. It has a low complication rate, high patient satisfaction, and excellent esthetic results. PMID- 15786432 TI - Successful use of letrozole in male breast cancer: a case report and review of hormonal therapy for male breast cancer. AB - Male breast cancer is an uncommon occurrence. Treatment of male breast cancer is typically extrapolated from data on the treatment of female breast cancer. Recently, aromatase inhibition has been proven as an effective therapy for female breast cancer, particularly in the setting of advanced cancers. The efficacy of aromatase inhibitors in males, however, has not been established. We report the successful treatment of a male with locally advanced breast cancer using the aromatase inhibitor letrozole. PMID- 15786433 TI - Hepatectomy plus intraoperative radiofrequency ablation and chemotherapy to treat technically unresectable multiple colorectal liver metastases. AB - BACKGROUND: Results and indications of intra-operative radiofrequency (RF) ablation of liver metastases (LM) are not well defined in the literature. AIM: To appreciate the survival rate of patients with strictly unresectable LM (defined on technical but not oncological criteria) when undergoing liver resection plus RF, along with optimal systemic chemotherapy. PATIENTS AND METHODS: Sixty three patients with technically unresectable LM (either >5, or bilateral with no sparing of at least one sector of the liver, or with tumor proximity to central major vascular structures) were treated. Extrahepatic metastases were also resected in 27% of patients. All patients received perioperative chemotherapy. The median follow-up was 27.6 months (range: 15-74). RESULTS: There was no postoperative mortality and the morbidity rate was 27%. The 2-year overall survival rate of the 63 patients was 67% with a median survival of 36 months. The local recurrence rates were similar for the three types of local treatments: 7.1% for the 154 RF ablations, 7.2% for the 55 wedge resections, and 9% for the 44 segmental anatomic resections (P = 0.216). Hepatic recurrences occurred in 71% of patients. CONCLUSION: The combination of anatomic segmental and wedge resections, RF ablation, and optimal chemotherapy in patients with technically unresectable LM results in a median survival of 36 months. PMID- 15786434 TI - Extremity soft tissue sarcoma: evidence-based multidisciplinary management. PMID- 15786435 TI - Cortical bone screw fixation in ionically modified apatite cements. AB - Hydroxyapatite cements are used in reconstruction of the face; usually in well defined cavities where the cement can be stabilized without the need for internal fixation. A hydroxyapatite cement that could enable screw fixation and some loading therefore has considerable potential in maxillofacial reconstruction. It has been demonstrated recently that water demand of calcium phosphate cements can be reduced by ionically modifying the liquid component. This study investigated the capacity of an ionically modified precompacted apatite cement to retain self tapping cortical bone screws. Screw pullout forces were determined in the direction of the screw long axis and perpendicular to it, using cortical bone and polymethylmethacrylate cement as a control. In bending pullout tests, measured forces to remove screws from ionically modified precompacted cement were insignificantly different from cortical bone. However, pullout forces of bone screws from hydroxyapatite cement decreased with aging time in vitro. PMID- 15786436 TI - Fibrin adhesive implant in wound healing repair of dental sockets with topical application of epsilon aminocaproic acid: histological analysis. AB - The aim of the study was to evaluate wound healing repair of dental sockets after topical application of 5% epsilon-aminocaproic acid (EACA) and the use of fibrin adhesive implant in rats under anticoagulant therapy with warfarin. Sixty Albinus wistar rats were used, divided into three groups of 20. In Group I, the animals were given 0.1 mL/100 mg of 0.9% saline solution per day, beginning 6 days before dental extraction and continuing throughout the experimental period. In Group II, the animals received 0.03 mL of sodium warfarin daily, beginning 6 days before the surgery and continuing until the day of sacrifice; after tooth extractions, the sockets were filled with fibrin adhesive material. In Group III the animals were treated as in Group II, and after extractions, the sockets were irrigated with 5 mL of 5% EACA and filled with the same fibrin adhesive material. All groups presented biological phases of wound healing repair, the differences being evident only in the chronology. The results obtained in Group III were very similar to those of Group I in the last period of wound repair, whereas Group II presented a late chronology compared to the other groups. PMID- 15786437 TI - Pediatric lung disease: from proteinases to pulmonary fibrosis. AB - One distinctive outcome of interstitial lung diseases in childhood is the abnormal accumulation of pulmonary extracellular matrix. The clinical consequence of such excessive connective tissue accumulation is known as pulmonary fibrosis. While numerous aspects of its pathogenesis have become familiar, many key events involved in its inception and progression still remain unclear. There is now compelling evidence that lung damage due to uncontrolled proteolysis may help drive critical processes that regulate fibrotic matrix remodeling. In this regard, a number of proteinases have been implicated in promoting both the initial lung injury and the fibroproliferative repair that follows. This review summarizes the knowledge of how different matrix-targeting enzymes may act to influence the development of pediatric pulmonary fibrosis. Understanding the scientific basis of this complex process may highlight opportunities to limit unwanted proteolysis and the intensity of its fibrotic sequelae. PMID- 15786438 TI - Comparative study of SP[6-11] and its analogs using simulated annealing. AB - In this study we compared the steric structures of the bioactive part of substance P (SP[6-11]) and its analogs (NY3460 and pHOPA-SP5). The molecular dynamics-simulated annealing method was used to explore the conformational space, and the structural differences and similarities of these molecules were identified. For the three peptides, the conformational distributions were represented in Ramachandran density plots. The occurring secondary structural elements of the investigated molecules were identified, namely alpha-Helix, type III beta-Turn, gamma-Turn, and inverse gamma-Turn. For SP[6-11] and its two analogs, different intramolecular interactions (H-bonds between the main-chain atoms, aromatic-aromatic interactions, and amino-aromatic interactions) that can stabilize the various conformations of the three peptides were investigated. Detailed examination of these intramolecular interactions revealed that H-bonds between the main-chain atoms are relevant in the determination and stabilization of the conformer structures of the peptides, while the aromatic-aromatic interactions do not play an important stabilizing role. Furthermore, in the conformers of NY3460 and pHOPA-SP5, different types of amino-aromatic interactions were identified that contribute to the formation of the various structures of these peptides. For all three molecules, the orientations of the side chains were investigated and the rotamer populations were determined. PMID- 15786439 TI - Vascular endothelial growth factor in preterm infants with respiratory distress syndrome. AB - Respiratory distress syndrome (RDS) secondary to surfactant deficiency is a common cause of morbidity and mortality in premature infants. Increasing evidence suggests that vascular endothelial growth factor (VEGF) may contribute to surfactant secretion and pulmonary maturation. However, differences in cord blood VEGF concentrations in infants with and without respiratory distress syndrome have not been reported. We hypothesized that premature infants with higher VEGF levels in cord blood had a lower risk of developing RDS. Cord blood samples were obtained from preterm infants born at 32 weeks of gestation or earlier. Infants were excluded if there was evidence of prenatal maternal infection or any infection within the first 3 days of life. Cord blood VEGF levels were measured using an enzyme-linked immunosorbent assay (ELISA). We found that neonates with clinically diagnosed RDS had a lower gestational age (GA), lower birth weight (BW), higher incidence of mechanical ventilation requirements, longer duration of mechanical ventilation, and lower Apgar scores at 1 and 5 min. Infants with RDS had significantly lower cord blood VEGF levels. GA, BW, premature rupture of membranes (PROM), and antenatal steroid treatment were not associated with changes in cord blood VEGF levels. The specificity of cord blood VEGF above 34 pg/ml for predicting the absence of RDS was 86%, the sensitivity was 53%, the positive predictive value was 84%, and the negative predictive value was 56%. Our data demonstrated that cord blood VEGF elevation was significantly correlated with an absence of RDS. PMID- 15786441 TI - Pulmonary sarcoidosis in a child with cystic fibrosis. AB - The co-occurrence of cystic fibrosis (CF) and sarcoidosis is rare. This case report describes a patient with both diseases. A 13-year-old girl with known CF (homozygous delta F508 defect) presented with a sudden decline in lung function. FEV1 decreased from 80% to 64% predicted, and FVC from 90% to 80% predicted. Diffusion capacity for carbon monoxide (D(L)CO) was 92% predicted. There was no history of cough, dyspnea, or reduced exercise tolerance, but she had arthralgia of the knee- and ankle-joints. A chest radiograph and CT scan of the thorax demonstrated pronounced bilateral hilar and mediastinal lymphadenopathy, compatible with pulmonary sarcoidosis. Histological examination of lymph node biopsy specimens obtained at mediastinoscopy demonstrated noncaseating epithelioid-cell granuloma. The majority of lymphocytes were CD4+ T lymphocytes, with a CD4+/CD8+ ratio of 5:1. The patient showed a prompt response to treatment with oral corticosteroids, and lung function returned to baseline levels. Subsequent radiographic appearances showed almost complete regression of mediastinal lymphadenopathy. The probability that CF and sarcoidosis would coexist by chance in a Danish child of this age is approximately 1:10(9). The collective incidence and geographic distribution of previously described patients with coexistent CF and sarcoidosis lend support to an association between the two diseases. PMID- 15786440 TI - Randomized controlled trial of salbutamol aerosol therapy via metered dose inhaler-spacer vs. jet nebulizer in young children with wheezing. AB - The jet nebulizer is a common device used for administering aerosol medication in young children. However, compared to a metered dose inhaler-spacer (MDI-spacer), it takes more time and personnel. This study aimed to compare the efficacy of salbutamol aerosol therapy given via these two devices in young wheezing children. A prospective randomized, double-blind, placebo-controlled trial was performed in children up to 5 years old who had acute wheezing and were admitted to the Department of Pediatrics, King Chulalongkorn Memorial Hospital. Patients were randomly divided into two groups. The first group received 2 puffs of placebo via MDI-spacer, followed by 0.15 mg/kg salbutamol respiratory solution via jet nebulizer. The second group received 2 puffs (100 microg/puff) of salbutamol via MDI-spacer, followed by placebo via jet nebulizer. Clinical scores and tidal breathing pulmonary function test were evaluated before and after treatment. Pulmonary function parameters included those derived from flow volume loops (volume to peak tidal expiratory flow over total expiratory volume, V(PTEF)/V(E); time to peak tidal expiratory flow over total expiratory time, T(PTEF)/T(E); and ratio of tidal expiratory flow at 25% remaining expiration to peak expiratory flow, 25/PF), compliance (Crs), and resistance (Rrs) of the respiratory system. The efficacy of both methods was compared by using analysis of covariance. Forty-seven wheezing children were studied (24 received salbutamol via MDI-spacer, and 23 received it via jet nebulizer). There was no statistical difference between the two groups regarding clinical scores and all pulmonary function parameters. However, heart rate was significantly increased after treatment in the jet nebulizer group when compared to those in the MDI-spacer group (P = 0.004). In conclusion, the efficacy of salbutamol aerosol therapy via MDI-spacer compared to jet nebulizer in young wheezing children was not different in terms of clinical score and postbronchodilator pulmonary function parameters. However, salbutamol aerosol therapy via jet nebulizer significantly increased the heart rate when compared to the MDI-spacer. PMID- 15786442 TI - Natural history of snoring and obstructive sleep apnea in Thai school-age children. AB - In 1999, a survey was carried out in 1,008 Thai children aged 7 years, which found that 85 (8.5%) children were habitual snorers, and 7 (0.69%) children had mild obstructive sleep apnea syndrome (OSAS). Since the natural history of snoring and untreated mild OSAS is still largely unknown, this study was undertaken in 2002 in the same group of children to determine the natural history of snoring and OSAS. Questionnaires, consisting of questions about snoring, were sent to the parents of the 1,008 children. Polysomnography was performed in 1) the 7 children who had OSAS in the previous survey, and 2) other habitual snorers who had sleep-related symptoms in this survey. Seventy-five percent of the questionnaires were returned. The prevalence of habitual snoring had decreased slightly, from 8.5% in 1999 to 6.9% in 2002. Sixty-five percent of the children who had snored habitually in the previous survey no longer did so, whereas 4.5% of the children who previously never snored or snored sometimes had become habitual snorers. Of the 7 children who had OSAS previously, 5 had persistent snoring, and polysomnographic studies revealed more severe OSAS, with an apnea hypopnea index (AHI) of 1.5-9.2 per hour of sleep. Five children were newly diagnosed with OSAS in this survey, with an AHI of 1.5-7.5. The overall prevalence of OSAS in this survey was 10/755 (1.3%). In conclusion, 65% of children who snored habitually no longer did so when they got older, while 9% of children had developed OSAS. We suggest that regular follow-up in children with habitual snoring may be needed, and additional research is required to determine the indications for polysomnography and neurobehavioral and cardiovascular assessment. We also showed that children with mild OSAS could develop more severe disease if left untreated, suggesting that deferment of treatment may have negative consequences. PMID- 15786443 TI - Identification of hippocampus-related candidate genes for Alzheimer's disease. AB - Alzheimer's disease (AD) is a complex multifactorial disease in which many genetic and environmental factors are involved. We performed an association study using 376 AD patients and 376 control subjects. We studied 35 single nucleotide polymorphisms in 35 genes that were significantly downregulated or upregulated only in the AD hippocampus compared with control and found that 9 single nucleotide polymorphisms were associated with AD. Our data indicated that single nucleotide polymorphisms could highly reflect differences in gene expression. Furthermore, an intronic polymorphism (+9943T/C) in POU2F1 was most significantly associated with AD (p = 0.0007). Our results suggest that POU2F1 is a candidate gene for AD. PMID- 15786444 TI - Interferon-beta1a therapy in human T-lymphotropic virus type I-associated neurologic disease. AB - Human T-lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is an immune-mediated inflammatory disorder of the central nervous system. Immune activation in the host, which results from high levels of persistent antigenic stimulation and from transactivation of host immunoregulatory genes by HTLV-I, appears important in the pathogenesis of HAM/TSP. In a single-center, open-label trial, 12 patients with HAM/TSP were treated with doses of interferon-beta1a of up to 60mug twice weekly, based on its antiviral and immunomodulatory effects. Primary end points were immunological and virological measures that are potential biomarkers for HAM/TSP. Interferon-beta1a therapy reduced the HTLV-I tax messenger RNA load and the frequency of potentially pathogenic HTLV-I-specific CD8(+) cells. The HTLV-I proviral DNA load remained unchanged. Spontaneous lymphoproliferation, a marker of T-cell activation in HAM/TSP, also was reduced. Some measures of motor function were improved, and no significant clinical progression occurred during therapy. These results indicate that interferon-beta1a may beneficially affect the immune mechanisms central to the pathogenesis of HAM/TSP. PMID- 15786445 TI - Tape-casting technique can prepare beta-TCP sheets with uniform thickness and flexibility. AB - The objective of this study is to propose a new fabrication technology for bone substitutes. In this study, a tape-casting method was used to prepare flexible beta-tricalcium phosphate (beta-TCP) sheets. A beta-TCP slurry containing a binder and plasticizer was used in a doctor blade system. The beta-TCP sheet obtained by this tape-casting method was highly flexible, enabling twisting and free-form shaping. The beta-TCP sheet was approximately 0.21 mm thick. X-ray diffraction and Fourier transform infrared spectrometry revealed that the structure of the beta-TCP component in the sheet is the same as that of the original beta-TCP powder. Observation by field-emission scanning electron microscopy showed that the beta-TCP sheet had a flat, microgranular surface. During the early stages, the tensile stress-strain curves of the beta-TCP sheet showed a nonlinear behavior until reaching the point of final fracture. This result was derived from the ductile property of the prepared beta-TCP sheet. In conclusion, a flexible beta-TCP sheet was easily prepared using a tape-casting technique. Fabrication using tape casting offers the advantages of enabling the preparation of ceramic sheets with precise thickness and not requiring expensive fabrication facilities. PMID- 15786446 TI - Creatine metabolism in combined methylmalonic aciduria and homocystinuria. AB - Methylation is an important aspect of many fundamental biological processes including creatine biosynthesis. We studied five patients with an inborn error of cobalamin metabolism to characterize the relation between homocysteine and creatine metabolism. Plasma guanidinoacetate concentrations were increased, 14.9 +/- 4.8 micromol/L (p < 0.0001), whereas plasma creatine concentrations were in the low reference range, 43.8 +/- 20.7 micromol/L (p = not significant). Individuals with combined methylmalonic aciduria and homocystinuria have a functional impairment of the creatine synthetic pathway probably secondary to a relative depletion of labile methyl groups. The neurotoxic effects of guanidinoacetate may be partly responsible for the observed neurological phenotype. PMID- 15786447 TI - Influence of the radiopacifier in an acrylic bone cement on its mechanical, thermal, and physical properties: barium sulfate-containing cement versus iodine containing cement. AB - In all acrylic bone cement formulations in clinical use today, radiopacity is provided by micron-sized particles (typical mean diameter of between about 1 and 2 microm) of either BaSO(4) or ZrO(2). However, a number of research reports have highlighted the fact that these particles have deleterious effects on various properties of the cured cement. Thus, there is interest in alternative radiopacifiers. The present study focuses on one such alternative. Specifically, a cement that contains covalently bound iodine in the powder (herein designated the I-cement) was compared with a commercially available cement of comparable composition (C-ment3), in which radiopacity is provided by BaSO(4) particles (this cement is herein designated the B-cement), on the basis of the strength (sigma(b)), modulus (E(b)), and work-to-fracture (U(b)), under four-point bending, plane-strain fracture toughness (K(IC)), Weibull mean fatigue life, N(WM) (fatigue conditions: +/-15 MPa; 2 Hz), activation energy (Q), and frequency factor (ln Z) for the cement polymerization process (both determined by using differential scanning calorimetry at heating rates of 5, 10, 15, and 20 K min( 1)), and the diffusion coefficient for the absorption of phosphate-buffered saline at 37 degrees C (D). For the B-cement, the values of sigma(b), E(b), U(b), K(IC), N(WM), Q, ln Z, and D were 53 +/- 3 MPa, 3000 +/- 120 MPa, 108 +/- 15 kJ m(-3), 1.67 +/- 0.02 MPa check mark m, 7197 cycles, 243 +/- 17 kJ mol(-1), 87 +/- 6, and (3.15 +/- 0.94) x 10(-12) m(2) s(-1), respectively. For the I-cement, the corresponding values were 58 +/- 5 MPa, 2790 +/- 140 MPa, 118 +/- 45 kJ m(-3), 1.73 +/- 0.11 MPa check mark m, 5520 cycles, 267 +/- 19 kJ mol(-1), 95 +/- 9, and (3.83 +/- 0.25) x 10(-12) m(2) s(-1). For each of the properties of the fully cured cement, except for the rate constant of the polymerization reaction, at 37 degrees C (k'), as estimated from the Q and ln Z results, there is no statistically significant difference between the two cements. k' for the I-cement was about a third that for the B-cement, suggesting that the former cement has a higher thermal stability. The influence of various characteristics of the starting powder (mean particle size, particle size distribution, and morphology) on the properties of the cured cements appears to be complex. When all the present results are considered, there is a clear indication that the I-cement is a viable candidate cement for use in cemented arthroplasties in place of the B cement. PMID- 15786448 TI - Functionalization of dental implant surfaces using adhesion molecules. AB - The aim of the present study was to test the hypothesis that organic coating of titanium screw implants that provides binding sites for integrin receptors can enhance periimplant bone formation. Ten adult female foxhounds received experimental titanium screw implants in the mandible 3 months after removal of all premolar teeth. Four types of implants were evaluated in each animal: (1) implants with machined titanium surface, (2) implants coated with collagen I, (3) implants with collagen I and cyclic RGD peptide coating (Arg-Gly-Asp) with low RGD concentrations (100 micromol/mL), and (4) implants with collagen I and RGD coating with high RGD concentrations (1000 micromol/mL). Periimplant bone regeneration was assessed histomorphometrically after 1 and 3 months in five dogs each by measuring bone implant contact (BIC) and the volume density of the newly formed periimplant bone (BVD). After 1 month, BIC was significantly enhanced only in the group of implants coated with the higher concentration of RGD peptides (p = 0.026). Volume density of the newly formed periimplant bone was significantly higher in all implants with organic coating. No significant difference was found between collagen coating and RGD coatings. After 3 months, BIC was significantly higher in all implants with organic coating than in implants with machined surfaces. Periimplant BVD was significantly increased in all coated implants in comparison to machined surfaces also. It was concluded that organic coating of machined screw implant surfaces providing binding sites for integrin receptors can enhance bone implant contact and periimplant bone formation. PMID- 15786449 TI - Acute intermittent porphyria presenting as a diffuse encephalopathy. AB - Although acute intermittent porphyria presents with dramatic neurological findings, the diagnosis is difficult. An 18-year-old woman had a clinical picture of porphyric encephalopathy. Magnetic resonance (MR) imaging demonstrated multiple large contrast-enhancing subcortical white matter lesions, which regressed with glucose and hematin infusions. Diffusion-weighted MR imaging was normal, and MR spectroscopy excluded acute demyelination or tissue necrosis. MR findings of acute intermittent porphyria can differ from those in posterior reversible encephalopathy syndrome by virtue of intense contrast enhancement. Because diffusion-weighted MR imaging and spectroscopy were normal, the lesions are likely caused by reversible vasogenic edema and transient breakdown of the blood-brain barrier. PMID- 15786451 TI - Fright is a provoking factor in vanishing white matter disease. AB - Leukoencephalopathy with vanishing white matter is an inherited disorder with a chronic progressive disease course and additional episodes of rapid neurological deterioration. These episodes typically are provoked by febrile infections or minor head trauma. We report on two patients who experienced an episode of rapid neurological deterioration after a fright. PMID- 15786452 TI - Dilatative arteriopathy (dolichoectasia): What is known and not known. PMID- 15786453 TI - Antemortem diagnosis of frontotemporal lobar degeneration. AB - The objective of this article is to study the accuracy of antemortem clinical diagnoses of frontotemporal lobar degenerations (FTLDs). From brain autopsies performed on subjects enrolled in the Mayo Alzheimer Center between 1991 and 2003, cases with neuropathological diagnoses of FTLD were identified. Neuropathological diagnoses of FTLDs were based on consensus criteria for FTLD. The initial clinical histories, neuropsychological test results, brain imaging studies, and initial clinical diagnoses were reviewed. There were 34 pathological FTLD cases among 433 subjects who underwent autopsy; 29 of these 34 cases were diagnosed as FTLD antemortem based on the sum of clinical, neuropsychological, and imaging features (sensitivity, 85%). The specificity was 99%. Among the 34 cases with pathological FTLD, 27 (79%) had clinical histories diagnostic of an FTLD syndrome, 20 (62%) had neuropsychological profiles consistent with FTLD, 17 (50%) had magnetic resonance scans consistent with FTLD, and 7 of 8 who had functional imaging studies had ones consistent with FTLD. In those with incorrect antemortem diagnoses, three were thought to have Alzheimer's disease, one was considered hard to classify, and one was diagnosed with vascular dementia. The antemortem consensus diagnosis of FTLD was moderately sensitive and very specific. With experienced clinicians and awareness of the unique manifestations of FTLD, accurate antemortem diagnosis was feasible. PMID- 15786455 TI - The United States' share of published research in neurosciences and clinical neurology. PMID- 15786454 TI - The metabolic pathology of dopa-responsive dystonia. AB - We used [(18)F]-fluorodeoxyglucose and positron emission tomography to determine a discrete cerebral pattern of abnormal glucose utilization in dopa-responsive dystonia. Network analysis demonstrated that dopa-responsive dystonia is associated with a specific pattern of regional metabolic covariation, characterized by increases in the dorsal midbrain, cerebellum, and supplementary motor area, as well as reductions in motor and lateral premotor cortex and in the basal ganglia. This pattern was not expressed in mutation carriers for primary torsion dystonia. Dopa-responsive dystonia has a unique metabolic architecture that differs from other inherited forms of dystonia. PMID- 15786456 TI - Plasma testosterone in male patients with Huntington's disease: relations to severity of illness and dementia. AB - Huntington's disease (HD) is a neurodegenerative disorder characterized by motor, cognitive, and psychiatric symptoms and by a progressive loss among other, of dopaminergic receptors in striatum, cortex, and hypothalamus. Central dopaminergic activity has been implicated in the regulation of sex hormones. Several features of testosterone deficiency, such as reduced muscle mass, depressive mood, and cognitive impairment, are often present in HD patients, but data on their testosterone levels are lacking. We assessed plasma levels of testosterone, LH, and FSH in 42 male patients with HD, confirmed by molecular genetic analysis, and searched for differences from age-matched healthy male subjects and for relations to CAG repeat number, age, age range, 26 to 76 (mean, 50.7 +/- 12.3) years; duration of illness range, 1 to 23 (mean, 6.7 +/- 6.3) years; and CAG repeat numbers from 40 to 65 (45.1 +/- 3.8). Disease symptomatology was assessed using the Unified Huntington's Disease Rating Scale. Testosterone and LH levels of the patients were significantly lower compared to the levels of 44 age-matched (mean age, 48.9 +/- 13.0, range, 26-76 years) healthy men. Severity of illness was negatively related to plasma testosterone levels. Further, low testosterone levels were associated with dementia but not with depression or psychotic features. Clinical studies with selected HD patients are needed to evaluate possible beneficial effects of androgen substitution therapy on cognitive functions, depression, muscle mass and strength, general well-being, and, eventually, neuroprotective effects. PMID- 15786457 TI - Nogo expression in muscle correlates with amyotrophic lateral sclerosis severity. AB - Nogo, a protein inhibiting axonal regeneration, exhibits a characteristic isoform specific pattern of expression in skeletal muscle of transgenic mice and patients with amyotrophic lateral sclerosis. Here, the increased levels of Nogo-A or Nogo B in muscle biopsies of 15 amyotrophic lateral sclerosis patients significantly correlated with the severity of clinical disability and with the degree of muscle fiber atrophy. Nogo-A immunoreactivity was observed selectively in atrophic slow twitch type I fibers. These results suggest that Nogo expression in muscle is a marker of amyotrophic lateral sclerosis severity. PMID- 15786460 TI - How to use spinal cord magnetic resonance imaging in the McDonald diagnostic criteria for multiple sclerosis. PMID- 15786459 TI - Metabolic penumbra of acute brain infarction: a correlation with infarct growth. AB - Volume expansion associated with brain infarction occurs in perfusion-diffusion mismatch of magnetic resonance imaging. We aimed at elucidating the metabolic impairment of this phenomenon with (15)O positron emission tomography and perfusion and diffusion magnetic resonance imaging. Eleven patients with acute unilateral embolic occlusion of the internal carotid or middle cerebral artery were studied within 6 hours of onset. Regional cerebral blood flow and cerebral metabolic rate of oxygen (CMRO(2)) were compared with those in the contralateral cerebral hemisphere. The relative apparent diffusion coefficient of water was estimated as a marker of cytotoxic edema. Relative cerebral blood flow and relative CMRO(2) in an evolving infarct (normal diffusion initially, but abnormal on day 3) were significantly (p < 0.05) less than those in the periinfarct area (normal diffusion initially and on day 3). The relative apparent diffusion coefficient between the evolving infarct and periinfarct showed no significant difference. These findings indicated that the initial 3-day volume expansion of an embolic brain infarction was associated with disturbed CMRD(2) but not with cytotoxic edema as early as 6 hours after onset. The "metabolic penumbra" defined as normal water diffusion with depressed CMRO(2) is a target to reduce the volume expansion of brain infarction. PMID- 15786462 TI - Early onset neuropathy in a compound form of Charcot-Marie-Tooth disease. AB - A 2-year-old boy presented with early-onset Charcot-Marie-Tooth disease (CMT). His parents had not been diagnosed previously with CMT, but on careful examination they showed clinical signs of CMT and reduced nerve conduction velocities. Genetic analysis identified the boy as a heterozygote for both a peripheral myelin protein 22 (PMP22) duplication and a mutation in the lipopolysaccharide-induced-tumour-necrosis-factor-alpha-factor (LITAF) gene, whereas each parent only had one mutated CMT gene. This suggests that LITAF mutations can severely affect the CMT phenotype caused by a PMP22 duplication. PMID- 15786463 TI - Commonality of TRIM32 mutation in causing sarcotubular myopathy and LGMD2H. AB - Sarcotubular myopathy (OMIM 268950) is a rare autosomal recessive myopathy first described in two Hutterite brothers from South Dakota and in two non-Hutterite brothers from Germany. We report that sarcotubular myopathy (STM) is caused by mutation in TRIM32, the gene encoding the tripartite motif-containing protein 32. TRIM32 was found to be the gene mutated in limb girdle muscular dystrophy type 2H (LGMD2H [OMIM 254110]), a disorder that has been confined to the Hutterite population. The TRIM32 mutation found in the STM patients is identical to the causative mutation for LGMD2H (D487N), Haplotype analysis shows that the disease chromosomes share common ancestry. PMID- 15786464 TI - Mapping of a new form of pure autosomal recessive spastic paraplegia (SPG28). AB - Pure hereditary spastic paraplegias are characterized by isolated and progressive spasticity in the lower limbs. We mapped the spastic paraplegia 28 (SPG28) locus to chromosome 14q21.3-q22.3 in a Moroccan family with autosomal recessive hereditary spastic paraplegia. Affected patients experienced development of progressive spastic gait during childhood and required help walking in their early 40s. Nine additional hereditary spastic paraplegia families were not linked to this locus, demonstrating further genetic heterogeneity. No mutations were found in exons of GCH1 and SPG3A, two genes from the candidate region involved in movement disorders. PMID- 15786465 TI - Normobaric hyperoxia extends the reperfusion window in focal cerebral ischemia. AB - A major limitation in thrombolysis for acute ischemic stroke is the restricted time window for safe and effective therapy. Any method that can extend the reperfusion time window would be important. In this study, we show that normobaric hyperoxia extends the time window for effective reperfusion from 1 to 3 hours in rats subjected to focal cerebral ischemia. Normobaric hyperoxia did not increase cellular markers of superoxide generation or brain levels of matrix metalloproteinase-9. Normobaric hyperoxia may be a clinically feasible adjunct method for significantly increasing the time window for effective thrombolytic therapy in acute ischemic stroke. PMID- 15786466 TI - JC virus granule cell neuronopathy: A novel clinical syndrome distinct from progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML) typically affects the CNS white matter of the central nervous system. We present an human immunodeficiency virus infected patient with polyomavirus JC infection restricted to granule cell neurons of the cerebellum and with corresponding neurological symptomatology. Magnetic resonance imaging demonstrated cerebellar atrophy without white matter lesions and stereotactic biopsy showed selective infection of the cerebellar granular cell layer, with preservation of Purkinje cells and absence of classic progressive multifocal leukoencephalopathy histopathology in underlying white matter. Evolution over 8 years was marked by symptomatic improvement corresponding to highly active antiretroviral therapy (HAART), with modest increase in CD4(+) T-cell counts. We propose to call this novel syndrome JCV granule cell neuronopathy (JCV GCN). PMID- 15786469 TI - Mitochondrial DNA haplogroup cluster UKJT reduces the risk of PD. AB - There is increasing evidence that genetic variants of mitochondrial DNA have an important role in the cause of idiopathic Parkinson's disease. We determined the mitochondrial DNA haplogroup of 455 Parkinson's disease cases, 185 Alzheimer's disease cases, and 447 healthy English control subjects. The UKJT haplogroup cluster was associated with a 22% reduction in population-attributable risk for Parkinson's disease. There was no association between individual haplogroups or the UKJT cluster and Alzheimer's disease, confirming that the association with Parkinson's disease was disease specific and not a general effect seen in all neurodegenerative diseases. PMID- 15786467 TI - Multiple regions of alpha-synuclein are associated with Parkinson's disease. AB - alpha-Synuclein is considered to play an important role in the pathogenesis of both the rare familial and the common sporadic forms of Parkinson's disease. Previous reports primarily have tested the association of alpha-synuclein promoter polymorphisms with idiopathic Parkinson's disease, but results are controversial. We first characterized the linkage disequilibrium structure of the alpha-synuclein gene region with a dense set of 56 genetic markers and subsequently performed two independent case-control association analyses using tagging markers. We could distinguish two large linkage disequilibrium blocks spanning the alpha-synuclein gene. Several markers within the 3'-block around exons 5 and 6 showed strong association with Parkinson's disease (p = 0.00009). Effects of the associated variants might be mediated by regulatory elements in this highly conserved region or by a frequency shift in a previously described splice variant lacking exon 5. A direct association with promoter polymorphisms could not be replicated in our sample set. A second set of markers in the 5' block of the gene were also significantly associated with Parkinson's disease, when young patients and female subjects were analyzed separately. These findings indicate locus heterogeneity for the pathogenesis of Parkinson's disease in different genetic or physiological environments, related to sex and age. PMID- 15786470 TI - Interleukin-1beta and febrile seizures: from bench to bedside. PMID- 15786473 TI - Upbeat nystagmus as the initial clinical sign of Creutzfeldt-Jakob disease. PMID- 15786474 TI - Convection perfusion of glucocerebrosidase for neuronopathic Gaucher's disease. AB - Systemic enzyme replacement for Gaucher's disease has not prevented premature death or severe morbidity in patients with a neuronopathic phenotype, because the enzyme does not cross the blood-brain barrier. We used convection-enhanced delivery for regional distribution of glucocerebrosidase in rat and primate brains and examined its safety and feasibility for neuronopathic Gaucher's disease. Rats underwent intrastriatal infusion and were observed and then sacrificed at 14 hours, 4 days, or 6 weeks. Primates underwent serial magnetic resonance imaging during enzyme perfusion of the right frontal lobe or brainstem, were observed and then sacrificed after infusion completion. Animals underwent histologic and enzymatic tissue analyses. Magnetic resonance imaging revealed perfusion of the primate right frontal lobe or pons with infusate. Enzyme activity was substantially and significantly (p < 0.05) increased in cortex and white matter of the infused frontal lobe and pons compared to control. Immunohistochemistry demonstrated intraneuronal glucocerebrosidase. There was no toxicity. Convection-enhanced delivery can be used to safely perfuse large regions of the brain and brainstem with therapeutic levels of glucocerebrosidase. Patients with neuronopathic Gaucher's disease and similar central nervous system disorders may benefit from this treatment. PMID- 15786475 TI - Murine gammaherpesvirus-68 infection of mice: A new model for human cerebral Epstein-Barr virus infection. AB - Epstein-Barr virus infection may cause severe neurological complications that are not mirrored by animal models. Here, we show that nasal inoculation of newborn BALB/c wild-type mice with MHV-68, a murine gammaherpesvirus, causes cerebral infection with inflammation in 50% of the animals. The inflammatory patterns are strikingly similar to those known from Epstein-Barr virus, including hydrocephalus, meningitis, cerebellitis, focal or diffuse encephalitis, and temporal lobe encephalitis. This offers a new powerful tool to study the virological and immunological characteristics of cerebral gammaherpesvirus infections. PMID- 15786477 TI - Distinguishing the four genetic causes of Jouberts syndrome-related disorders. AB - Jouberts syndrome-related disorders are a group of recessively inherited conditions showing cerebellar vermis hypoplasia and the molar tooth sign of the midbrain-hindbrain junction. Recent analyses have suggested at least three loci, JBTS1 (9q34.3), -2 (11p11.2-q12.3), and -3 (6q23), but the phenotypic spectrum associated with each locus has not been delineated. In addition, deletions of the NPHP1 gene, usually responsible for isolated juvenile nephronophthisis, are occasionally encountered among Jouberts syndrome-related disorder patients. Here, we describe four novel families showing evidence of linkage to two of these loci, provide a 3.6Mb refinement of the JBTS2 locus, and perform a detailed comparison of all linked families identified so far, to define the clinical and radiographical hallmarks for each genetic condition. We find that JBTS1 and -3 primarily show features restricted to the central nervous system, with JBTS1 showing largely pure cerebellar and midbrain-hindbrain junction involvement, and JBTS3 displaying cerebellar, midbrain-hindbrain junction, and cerebral cortical features, most notably polymicrogyria. Conversely, JBTS2 is associated with multiorgan involvement of kidney, retina, and liver, in addition to the central nervous system features, and results in extreme phenotypic variability. This provides a useful framework for genetic testing strategies and prediction of which patients are most likely to experience development of systemic complications. PMID- 15786478 TI - Alpha-synuclein aggregation and its relation to neurodegenerative diseases. PMID- 15786476 TI - Detection of entorhinal layer II using 7Tesla [corrected] magnetic resonance imaging. AB - The entorhinal cortex lies in the mediotemporal lobe and has major functional, structural, and clinical significance. The entorhinal cortex has a unique cytoarchitecture with large stellate neurons in layer II that form clusters. The entorhinal cortex receives vast sensory association input, and its major output arises from the layer II and III neurons that form the perforant pathway. Clinically, the neurons in layer II are affected with neurofibrillary tangles, one of the two pathological hallmarks of Alzheimer's disease. We describe detection of the entorhinal layer II islands using magnetic resonance imaging. We scanned human autopsied temporal lobe blocks in a 7T human scanner using a solenoid coil. In 70 and 100 microm isotropic data, the entorhinal islands were clearly visible throughout the anterior-posterior extent of entorhinal cortex. Layer II islands were prominent in both the magnetic resonance imaging and corresponding histological sections, showing similar size and shape in two types of data. Area borders and island location based on cytoarchitectural features in the mediotemporal lobe were robustly detected using the magnetic resonance images. Our ex vivo results could break ground for high-resolution in vivo scanning that could ultimately benefit early diagnosis and treatment of neurodegenerative disease. PMID- 15786480 TI - Treatment of port wine stains with photodynamic therapy, using pulsed dye laser as a light source, compared with pulsed dye laser alone: a pilot study. AB - BACKGROUND AND OBJECTIVES: Laser-induced photo thermal damage has been combined with photodynamic therapy (PDT) using a systemic photosensitiser to treat vascular lesions. The efficacy of PDT using systemic 5-aminolaevulinic acid (5 ALA) as the photosensitiser and pulsed dye laser (PDL) as the light source in port wine stains (PWS) is unknown. STUDY DESIGNS/MATERIALS AND METHODS: We conducted an internally controlled pilot study comparing the efficacy of PDT using PDL as a light source, to PDL alone in the treatment of PWS. RESULTS: The PWS improved slightly in all patients but no significant difference was found between the three treatment arms in terms of lesional lightening or incidence and severity of side effects. CONCLUSIONS: There was no evidence of increased efficacy of PDT using PDL as a light source compared to PDL alone. There was also no significant difference in adverse events. Further studies using different treatment regimens over longer periods of time may be warranted. PMID- 15786481 TI - Giant SCA8 alleles in nine children whose mother has two moderately large ones. AB - We report here a family in which each of nine children has inherited giant SCA8 CTG expansions from a homozygous mother who has two moderately large SCA8 CTG alleles. In contrast, three homozygous male individuals and a case of coexistence of two expansions of the FRDA gene and one of SCA8, all of them with moderately large alleles, have transmitted their respective SCA8 expanded alleles with minor changes, as usually occurs in heterozygous male transmissions. PMID- 15786482 TI - Characterization of measurement artefacts in fluoroptic temperature sensors: implications for laser thermal therapy at 810 nm. AB - BACKGROUND AND OBJECTIVES: Fluoroptic sensors are used to measure interstitial temperatures but their utility for monitoring laser interstitial thermal therapy (LITT) is unclear because these sensors exhibit a measurement artefact when exposed to the near-infrared (NIR) treatment light. This study investigates the cause of the artefact to determine whether fluoroptic sensors can provide reliable temperature measurements during LITT. STUDY DESIGN/MATERIALS AND METHODS: The temperature rise measured by a fluoroptic sensor irradiated in non absorbing media (air and water) was considered an artefact. Temperature rise was measured as a function of distance from a laser source. Two different sensor designs and several laser powers were investigated. A relationship between fluence rate and measurement artefact in water was determined and coupled with a numerical simulation of LITT in liver to estimate the error in temperature measurements made by fluoroptic sensors in tissue in proximity to the laser source. The effect of ambient light on the performance of sensors capped with a transparent material ("clear-capped sensors") was also investigated. RESULTS: The temperature rise recorded in air by both clear- and black-capped fluoroptic sensors decreased with distance from a laser source in a manner similar to fluence rate. Sensor cap material, laser power, and the thermal properties of the surrounding medium affected the magnitude of the artefact. Numerical simulations indicated that the accuracy of a clear-capped fluoroptic sensor used to monitor a typical LITT treatment in liver is > 1 degrees C provided the sensor is further than approximately 3 mm from the source. It was also shown that clear-capped fluoroptic sensors are affected by ambient light. CONCLUSIONS: The measurement artefact experienced by both black-capped and clear-capped fluoroptic sensors irradiated by NIR light scales with fluence rate and is due to direct absorption of the laser light, which results in sensor self-heating. Clear-capped fluoroptic sensors can be used to accurately monitor LITT in tissue but should be shielded from ambient light. PMID- 15786483 TI - Qualitative microanalysis of ions and ultrastructural changes in dentin exposed to laser irradiation and to metal salts solution. AB - BACKGROUND AND OBJECTIVES: This study evaluated the ultrastructural changes in dentin after treatment with the Nd:YAG laser and/or metal salt solutions and verified the presence of Sn++, Sr++, and F- in dentin structure. STUDY DESIGNS/MATERIALS AND METHODS: Sixty dentin disks were randomly divided into groups (n = 10): (I) control (no treatment), (II) Nd:YAG (1.5 W, 100 mJ, 15 Hz, 125 J/cm2), (III) 10% SnF2 aqueous solution for 30 minutes, (IV) Nd:YAG+10% SnF2 aqueous solution for 30 minutes, (V) 10% SrCl2 toothpaste for 30 minutes, (VI) Nd:YAG+10% SrCl2 toothpaste for 30 minutes. Then, all samples were prepared for scanning electron microscopy (SEM) and the samples from Groups I to IV for the energy dispersive X-ray microanalysis (EDX). RESULTS: SEM evaluation revealed occluded dentinal tubules and a dentin surface altered by the laser irradiation. The EDX microanalysis revealed Sn++ at a depth of 250 microm in Group IV and not deeper than 100 microm in Group III. In Group V, Sr++ was not deeper than 50 microm, but it could be detected at a depth of 500 microm in Group VI. F- was found only in Group IV. CONCLUSIONS: Ultrastructural changes caused by laser irradiation can increase dentin uptake of Sn++, Sr++, and F-. PMID- 15786484 TI - Anxiety disorder comobidity in Bipolar I Disorder: relationship to depression severity and treatment outcome. AB - The present study investigated the greater symptom severity and poorer treatment response found in patients with bipolar illness and anxiety comorbidity, and examined depression as a potential mediator of this relationship. The sample consisted of 92 patients in an acute episode of Bipolar I Disorder with a current or past history of an anxiety disorder. Diagnoses were based on structured clinical interview, and participants were assessed at pre-treatment and then randomly assigned to pharmacotherapy alone or pharmacotherapy plus family intervention. Patients were assessed on a monthly basis by blind assessors over 28 months. Compared to patients without anxiety comorbidity, individuals with bipolar disorder and an anxiety disorder possessed greater current symptom severity, even after controlling for depression severity. Logistic regression analysis identified that being female and having higher current depression but not manic severity predicted comorbid anxiety. Comorbid anxiety was associated with poorer treatment response in the sample regardless of treatment type, particularly in subsequent depressive symptoms. Multiple regression analyses indicated that current depression but not manic severity partially mediated the relationship between comorbid anxiety and treatment outcome. Results from the current study investigating comorbid anxiety disorders are consistent with past research limited to anxiety symptoms. Depression only partially accounted for the link between comorbid anxiety and greater symptom severity and poorer treatment response, and examination of other factors is warranted. Because of the clinical relevance of comorbid anxiety in severe affective disorders, treatments designed to specifically address both concerns are needed. PMID- 15786485 TI - Investigation of mood-congruent false and true memory recognition in depression. AB - The present study investigated the extent of mood-congruent false and true memory recognition in depression. A group of 25 patients with depression and 28 healthy controls completed a variant of the Deese-Roediger McDermott task. Four lists were read to participants in sequence, followed by a recognition task. The words in each list were associated with a central but unmentioned theme word that was either depression-relevant (i.e., loneliness), delusion-relevant (betrayal), positive (holidays), or neutral (window). Whereas it was expected to replicate the conventional mood-congruent effect in depression (better recognition of depression-relevant items), the available literature did not allow strong predictions to be made on the extent of mood-congruent false recognition in depression. Results showed that depressed patients learned emotionally charged material equally well as healthy participants but forgot significantly more neutral material. A conventional mood-congruent memory bias was not found, but relative to healthy controls, patients with depression committed more false recognition errors for emotionally charged words, particularly for depression relevant items. The results confirm that depressed patients are biased toward emotional material. Reasons for the absence of the expected mood-congruent memory bias are discussed. It is suggested that researchers as well as clinicians should pay more attention to mood-congruent false recollection, because it may undermine the validity of autobiographic reports in depressive patients and may represent a maintenance factor for the disorder. PMID- 15786487 TI - Escitalopram in clinical practice: results of an open-label trial in a naturalistic setting. AB - Results from randomized, placebo-controlled clinical trials have demonstrated that escitalopram is effective and well tolerated in the treatment of depression and anxiety disorders. Such trials typically employ stringent inclusion criteria that may limit the generalizability of findings to the broader population of patients treated in psychiatric and primary care practices. The objective of the current trial was to assess the effect of escitalopram treatment on depressed outpatients in naturalistic settings. This 8-week open-label trial enrolled 5,453 outpatients aged > or = 18 years with nonpsychotic major depressive disorder from primary care (n = 2,591), psychiatric (n = 2,289), and other specialty (n = 573) practices. Escitalopram was initiated at 10 mg/day, and dose could be increased to a maximum of 20 mg/day. Efficacy measures included the Clinical Global Impressions of Improvement (CGI-I) scale, Patient Global Evaluation (PGE) scale, Hospital Anxiety and Depression Scale, Sheehan Disability Scale, and 12-Item Short Form Health Survey. Overall, 76% of patients completed 8 weeks of treatment. The mean dose of escitalopram was 11.6 mg/day. At endpoint, response rates (defined as a score < or = 2 on the CGI-I or PGE) were 68% on the clinician assessed CGI-I and 66% on the PGE. Improvement was not related to age or response to prior antidepressant treatment. Overall, 9% of patients discontinued due to adverse events. Escitalopram treatment was well tolerated and associated with robust response rates in a broadly representative population of depressed outpatients. PMID- 15786486 TI - Potential effectiveness and safety of olanzapine in refractory panic disorder. AB - Panic disorder is a common and disabling psychiatric disorder. Despite treatment advances, refractory panic disorder requires novel interventions. One such pharmacologic intervention with theoretical and case study support includes olanzapine, a thienobenzodiazepine medication currently approved for schizophrenia in the United States. Ten people with refractory DSM-IV diagnosed panic disorder completed an 8-week, open-label, flexible-dose clinical trial. Baseline, in-treatment, and end-of-treatment data for panic attacks, anticipatory anxiety, phobic avoidance, and impairment were collected. Data were analyzed using SPSS software. Refractory panic disorder patients required a wide dose range averaging 12.3 mg/day of olanzapine to significantly improve or ablate panic attacks. On the average, number of attacks decreased from 6.1/week at baseline to 1.1/week at the end of treatment, and anticipatory anxiety from 32% of the day to 8% of the day. At treatment end, 5 of 10 participants (50%) were panic free, 4 (40%) had one attack in the previous week, 1 (10%) had seven attacks in the previous week, and 6 of 10 participants (60%) were anticipatory anxiety free. There were also statistically and clinically significant improvements in impairment over the course of the trial. There were no significant changes in vital signs, emergent side effects, or average weight, although 6 of 10 people did gain weight. Olanzapine is potentially effective and safe in panic disorder. Due to study limitations, further clinical trials are needed to demonstrate effectiveness. PMID- 15786488 TI - Correlation of in vitro chemopreventive efficacy data from the human epidermal cell assay with animal efficacy data and clinical trial plasma levels. AB - The human epidermal cell (HEC) assay, which uses carcinogen exposed normal skin keratinocytes to screen for cancer prevention efficacy, was used to screen possible preventive agents. The endpoints measured were inhibition of carcinogen induced growth and induction of involucrin, an early marker of differentiation. Sixteen of twenty agents (apigenin, apomine, budesonide, N-(2 carboxyphenyl)retinamide, ellagic acid, ibuprofen, indomethacin, melatonin, (-)-2 oxo-4-thiazolidine carboxylic acid, polyphenon E, resveratrol, beta-sitosterol, sulfasalazine, vitamin E acetate, and zileuton) were positive in at least one of the two assay endpoints. Four agents (4-methoxyphenol, naringenin, palmitoylcarnitine chloride, and silymarin) were negative in the assay. Nine of the sixteen agents were positive for both endpoints. Agents that showed the greatest response included: ellagic acid > budesonide, ibuprofen > apigenin, and quinicrine dihydrochloride. Fifty-eight of sixty-five agents that have been evaluated in the HEC assay have also been evaluated in one or more rodent bioassays for cancer prevention and several are in clinical trials for cancer prevention. The assay has an overall predictive accuracy of approximately 91.4% for efficacy in rodent cancer prevention irrespective of the species used, the tissue model, or the carcinogen used. Comparison of the efficacious concentrations in vitro to plasma levels in clinical trials show that concentrations that produced efficacy in the HEC assay were achieved in clinical studies for 31 of 33 agents for which plasma levels and/or C(max) levels were available. For two agents, 9-cis-retinoic acid (RA) and dehydroepiandrosterone (DHEA), the plasma levels greatly exceeded the highest concentration (HC) found to have efficacy in vitro. Thus, the HEC assay has an excellent predictive potential for animal efficacy and is responsive at clinically achievable concentrations. PMID- 15786489 TI - Hormonal regulation on regucalcin mRNA expression in cloned normal rat kidney proximal tubular epithelial NRK52E cells. AB - Regucalcin is a regulatory protein in cell signaling. This study was undertaken to determine whether regucalcin mRNA expresses in the cloned normal rat kidney proximal tubular epithelial NRK52E cells and its expression regulates due to hormones and cell signaling-related factors. Cells with subconfluency were cultured for 24, 48, or 72 h in a Dulbecco's modified Eagle medium supplemented with non-essential amino acid without bovine serum (BS). The result of Western blot analysis showed that regucalcin protein was present in the NRK52E cells. The expression of regucalcin mRNA in the cells was determined using reverse transcription-polymerase chain reaction (RT-PCR). Regucalcin mRNA expression in the NRK52E cells was significantly increased by culture with parathyroid hormone (PTH, 10(-8) or 10(-7) M), aldosterone (10(-8) or 10(-7) M), or dexamethasone (10(-8) M). The presence of 1,25-dihydroxyvitamin D(3) (1,25(OH)2D3, 10(-8) or 10(-7) M) or calcitonin (10(-9) or 10(-8) M) did not have a significant effect on regucalcin mRNA levels in the cells. Culture with dibutyryl cyclic AMP (DcAMP, 10(-5) or 10(-4) M) or phorbol 12-myristate 13-acetate (PMA, 10(-6) M), an activator of protein kinase C, caused a significant increase in regucalcin mRNA expression. The presence of staurosporine (10(-8) M) caused a significant decrease in regucalcin mRNA expression. Dibucaine (10(-7) M), PD98059 (10(-7) M), or vanadate (10(-6) or 10(-5) M) did not have an effect on regucalcin mRNA levels. The present study demonstrates that regucalcin mRNA and its protein are expressed in the cloned normal rat kidney proximal tubular epithelial NRK52E cells, and that the expression is enhanced by hormones which regulate ion transport in the proximal tubule. PMID- 15786490 TI - Osteogenic PTHs and vascular ossification-Is there a danger for osteoporotics? AB - Inflammation in vascular (mostly arterial) walls and heart valves triggered by the trans-endothelial influx of LDL particles and the action of subsequently modified (e.g., by oxidation) LDL particles can trigger true bone formation by valvar fibroblasts, by a subpopulation of re-differentiation-competent VSMCs (vascular smooth muscle cells) or by vascular pericytes. Vascular ossification can lead to heart failure and death. Elderly osteoporotic women who need osteogenic drugs to restore their lost skeletal bone are paradoxically prone to vascular ossification-the "calcification paradox." The recent introduction into the clinic of a potently osteogenic parathyroid hormone peptide, Lilly's rhPTH-(1 34)OH (Forteotrade mark), to reverse skeletal bone loss raises the question of whether this and other potently osteogenic PTHs still in clinical trial might also stimulate vascular ossification in such osteoporotic women. Indeed the VSMCs in human and rat atherosclerotic lesions hyperexpress PTHrP and the PTHR1 (or PTH1R) receptor as do maturing osteoblasts. And the evidence indicates that endogenous PTHrP with its NLS (nuclear/nucleolar localization sequence) does stimulate VSMC proliferation (a prime prerequisite for atheroma formation and ossification) via intranuclear targets that inactivate pRb, the inhibitory G1/S checkpoint regulator, by stimulating its hyperphosphorylation. But neither externally added full-length PTHrP nor the NLS-lacking PTHrP-(1-34)OH gets into the VSMC nucleus and instead they inhibit proliferation and calcification by only activating the cell's PTHR1 receptors. No PTH has an NLS and, as expected from the observations on the externally added PTHrPs, hPTH-(1-34)OH inhibits calcification by VSMCs and cannot stimulate vascular ossification in a diabetic mouse model. Encouraging though this may be for osteoporotics with their "calcification paradox," more work is needed to be sure that the skeletally osteogenic PTHs do not promote vascular ossification with its cardiovascular consequences. PMID- 15786491 TI - Regulation of skeletal development by the Runx family of transcription factors. AB - The Runx (runt-related protein) family of transcription factors plays important roles in different tissues and cell lineages. Runx1 determines commitment to the hematopoietic cell lineage and Runx2 determines commitment to the osteoblastic lineage. Cbfbeta is required for Runx1- and Runx2-dependent transcriptional regulation. Runx2 interacts with many other transcription factors and co regulators in the transcriptional regulation of its target genes. Runx2 is essential for the commitment of multipotent mesenchymal cells into the osteoblastic lineage and inhibits adipocyte differentiation. Runx2 induces the gene expression of bone matrix proteins, while keeping the osteoblastic cells in an immature stage. Runx2 and Runx3 have redundant functions in chondrocytes, and they are essential for chondrocyte maturation. Runx2 directly induces Indian hedgehog (Ihh) expression and co-ordinates the proliferation and differentiation of chondrocytes. Therefore, elucidation of the signaling pathways through Runx2 and Runx3 will unravel the complex mechanism of skeletal development. PMID- 15786492 TI - Stereotypical movements and frontotemporal dementia. AB - Stereotypical movements are characteristic of autism or mental retardation but can also occur in patients with dementia, particularly frontotemporal dementia (FTD). In this study, we administered the Abnormal Involuntary Movement Scale (AIMS) to 18 patients with FTD and to 18 patients with the most common form of dementia, Alzheimer's disease (AD). The AIMS scores were gathered at the initial presentation of patients who had not received antipsychotic medications. Compared to the AD patients, the FTD patients had significantly more stereotypical movements, including frequent rubbing behaviors and some self-injurious acts. All the FTD patients with stereotypical movements had compulsive-like behaviors, suggesting a similar pathophysiologic cause, and most had a decrease in their stereotypical movements with the administration of sertraline, a serotonin selective reuptake inhibitor. PMID- 15786493 TI - A COMPASS in the voyage of defining the role of trithorax/MLL-containing complexes: linking leukemogensis to covalent modifications of chromatin. AB - Chromosomal rearrangements and translocations play a major role in the pathogenesis of hematological malignancies. The trithorax-related mixed lineage leukemia (Mll) gene located on chromosome 11 is rearranged in a variety of aggressive human B and T lymphoid tumors as well as acute myeloid leukemia (AML) in both children and adults. It was first demonstrated for the yeast MLL homolog complex, Set1/COMPASS, and now for the MLL complex itself, that these complexes are histone methyltransferases capable of methylating the fourth lysine of histone H3. The post-translational modifications of histones by methylation have emerged as a key regulatory mechanism for both repression and activation of gene expression. Studies from several laboratories during the past few years have brought about a watershed of information defining the molecular machinery and factors involved in the recognition and modification of nucleosomal histones by methylation. In this review, we will discuss the recent findings regarding the molecular mechanism and consequences of histone modification by the MLL related protein containing complex COMPASS. PMID- 15786494 TI - Unintended pregnancies and exposure to potential human teratogens. AB - BACKGROUND: We aimed to investigate the risk factors associated with unintended pregnancies as well as the association between unintended pregnancies and potential teratogenic exposures. METHODS: A cross-sectional survey was performed among women attending the Maternity School of the Samsung Cheil Hospital and Women's Health Care Center in Seoul, Korea. Demographic data, obstetric history, socioeconomic status, intention to become pregnant, and exposure to potential teratogens were obtained. RESULTS: A total of 1354 women with median age of 29 years and median gestational age of 29 weeks were included. Of these, an educational level above high school was 74.2%, primigravida was 77.3% and unintended pregnancy was 48%. In the logistic regression analysis, women younger than 24 years of age had a relative risk (RR) of 2.5 (95% confidence interval [CI], 1.3-4.7) of having an unintended pregnancy and women with lower household monthly income level had a RR of 1.3 (95% CI, 1.0-1.6). Women with unintended pregnancies had an RR of 1.9 (95% CI, 1.5-2.5), 3.0 (95% CI, 2.0-4.5), 1.5 (95% CI, 1.0-2.3), 2.9 (95% CI, 1.1-7.2), and 2.0 (95% CI, 1.62.4) of being exposed to alcohol, medications, cigarette smoking, X-rays, or to any of these, respectively, during the first trimester of pregnancy. CONCLUSIONS: Unintended pregnancies are more likely to occur among young women with a lower household monthly income level. Prenatal counseling should be especially recommended for women with unintended pregnancies in order to evaluate whether they have been exposed to potential teratogenic agents. PMID- 15786495 TI - Kinetic expression of platelet endothelial cell adhesion molecule-1 (PECAM 1/CD31) during embryonic stem cell differentiation. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is widely used as a marker during vasculogenesis and angiogenesis from embryonic stem (ES) cells. However, the expression of PECAM-1 isoforms in ES cells has not been determined. The present study was designed to determine the role of PECAM-1 isoforms during in vitro endothelial differentiation of ES cells. It was found that undifferentiated ES cells expressed high level of PECAM-1, which primarily located at cell-cell junction, but the expression of PECAM-1 was sharply down regulated during early ES cell differentiation. In addition, undifferentiated ES cells were found the expressed all eight known alternatively spliced PECAM-1 isoforms, among them the expression of PECAM-1 isoforms lacking exon 15 or 14&15 was predominant. Quantitative analysis revealed a significant increase in the expression of PECAM-1 isoform lacking exon 12&14&15 as vascular development of ES cells. These results indicate a constitutive expression of PECAM-1 in undifferentiated murine ES cells and suggest a developmental role of PECAM-1 isoform changes during vasculogenesis and angiogenesis. PMID- 15786496 TI - Genetic polymorphisms: impact on the risk of fetal alcohol spectrum disorders. AB - Clinical reports on monozygotic and dizygotic twins provided the initial evidence for the involvement of genetic factors in risk vulnerability for fetal alcohol spectrum disorders (FASD) including fetal alcohol syndrome (FAS). Research with selectively bred and inbred rodents, genetic crosses of these lines and strains, and embryo culture studies have further clarified the role of both maternal and fetal genetics in the development of FASD. Research to identify specific polymorphisms contributing to FASD is still at an early stage. To date, polymorphisms of only one of the genes for the alcohol dehydrogenase enzyme family, the ADH1B, have been demonstrated to contribute to FASD vulnerability. In comparison with ADH1B*1, both maternal and fetal ADH1B*2 have been shown to reduce risk for FAS in a mixed ancestry South African population. ADH1B*3 appears to afford protection for FASD outcomes in African-American populations. Other candidate genes should be examined with respect to FASD risk, including those for the enzymes of serotonin metabolism, in particular the serotonin transporter. By its very nature, alcohol teratogenesis is the expression of the interaction of genes with environment. The study of genetic factors in FASD falls within the new field of ecogenetics. Understanding of the array of genetic factors in FASD will be enhanced by future genetic investigations, including case-control, family association, and linkage studies. PMID- 15786498 TI - Exploring SmBr2-, SmI2-, and YbI2-mediated reactions assisted by microwave irradiation. AB - The use of microwave heating in lanthanide(II) halide (LnX2 = SmBr2, SmI2, and YbI2) mediated reduction and coupling reactions has been investigated for a variety of functional groups including alpha,beta-unsaturated esters, aldehydes, ketones, imines, and alkyl halides. Good to quantitative transformations were obtained within a few minutes without the addition of any co-solvents, such as hexamethyl phosphoramide (HMPA). The redox potential of YbI2 in tetrahydrofuran (THF) has been determined as -1.02+/-0.05 V (versus Ag/AgNO3) by cyclic voltammetry. A large selectivity difference in various reactions was observed depending on the redox potential of the LnX2 reagent. The more powerful reductant, SmBr2, afforded mainly pinacol-coupling products of ketones whereas the weaker reductant YbI2 afforded mainly reduction products. The results indicate that the reducing power of LnX2 has a large impact on not only the pinacol coupling/reduction product ratio of ketones but also on other substrates in which there are competing coupling and reduction reactions. The use of in situ generated LnX2 has also been explored and proven useful in many of these reactions. PMID- 15786497 TI - Developmental abnormalities in rat embryos leading to tibial ray deficiencies induced by busulfan. AB - BACKGROUND: Little is known about the developmental changes associated with tibial ray deficiencies. The aim of this study was to detect cell death, proliferation, and gene expression that result in tibial ray deficiencies. METHODS: We induced tibial ray deficiencies in rat embryos using a teratogenic agent (busulfan) and observed the developmental changes in 1126 hindlimbs. We performed Nile blue staining, whole mount in situ hybridization for fibroblast growth factor 8 (Fgf8), bone morphogenetic protein 4 (Bmp4) and Sonic hedgehog (Shh), terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and assessment of cell proliferation by 5-bromo-2'-deoxy-uridine (BrdU)/anti-BrdU immunohistochemistry. RESULTS: In situ hybridization showed reductions in Fgf8 and Bmp4 expression. Histological examination showed a delay of mesenchymal condensation, increased mesenchymal cell death, decreased mesenchymal cell proliferation, and a reduction in the number of mesenchymal cells. These abnormalities may cause hypoplasia of the limb. Bmp4 expression was markedly reduced in the anterior mesenchyme. Shh was expressed in the posterior mesenchyme. We suggest that the posterior skeletal elements may be fully formed owing to Shh expression, but the anterior skeletal elements may be underdeveloped owing to an intense reduction of Bmp4 expression in the anterior mesenchyme, causing hypoplasia of the tibial ray. CONCLUSIONS: The combined effects of increased cell death, decreased cell proliferation, reduction of Fgf8 expression, and intense reduction of Bmp4 expression in the anterior mesenchyme may play an important role in the development of tibial ray deficiency induced by busulfan. PMID- 15786499 TI - Altered T cell differentiation and Notch signaling induced by the ectopic expression of keratin K10 in the epithelial cells of the thymus. AB - Transgenic mice expressing hK10 under the keratin K5 promoter display several alterations in the epidermis including decreased cell proliferation, and reduced susceptibility to tumor development. Given that K5 promoter is also active in the epithelial cells of the thymus, we explored the possible alterations of the thymus because of K10 transgene expression. We found severe thymic alterations, which affect not only the thymic epithelial cells (TEC), but also thymocytes. We observed altered architecture and premature thymus involution in the transgenic mice associated with increased apoptosis and reduced proliferation of the thymocytes. Interestingly, prior to the development of this detrimental phenotype, thymocytes of the transgenic mice also displayed altered differentiation, which is aggravated later on. Molecular characterization of this phenotype indicated that Akt activity is reduced in TEC, but not in thymocytes. In addition, we also observed altered expression of Notch family members and some of their ligands both in TEC and T cells. This produces reduced Notch activity in TEC but increased Notch activity in thymocytes, which is detectable prior to the disruption of the thymic architecture. In addition, we also detect altered Notch expression in the epidermis of bK5hK10 transgenic mice. Collectively the present data indicate that keratin K10 may induce severe alterations not only in a cell autonomous manner, but also in neighboring cells by the modulation of signals involved in cell-cell interactions. PMID- 15786501 TI - From a monomer to a protein-sized, doughnut-shaped coordination oligomer-the influence of side chains of C3-symmetric ligands in supramolecular chemistry. AB - Herein we describe the importance of side chains in C3-symmetric ligands in supramolecular chemistry. The reaction of the new ligand tris(5-bromo-2 methoxybenzylidene)triaminoguanidinium chloride [H3Me3Br3L]Cl (1) with ZnCl2 results in the formation of the monomeric complex (Et3NH)2[(ZnCl2)3Me3Br3L] (2), in which the ligand remains in a conformation less favourable for the coordination of metal centres. The use of the related tris(5-bromo-2 hydroxybenzylidene)triaminoguanidinium chloride, [H6Br3L]Cl, under similar conditions, results in the formation of two different dimeric compounds (NH4)[{[Zn(NH3)]3Br3L}2{mu-(OH)}3]1/4MeOH (3) and [Zn{Zn2(OH2)3(NH3)Br3L}2] (4), depending on the solvent mixture used. The comparable reaction of the ligand tris(5-bromo-2-hydroxy-3-methoxybenzylidene)triaminoguanidinium chloride [H6(OMe)3Br3L]Cl (5), leads to the formation of a doughnut-shaped, protein-sized coordination oligomer (Et3NH)18[{Zn[Zn2Cl{(OMe)3Br3L}]2}6(mu-Cl)6(OH2)6]x CH3CN (6), which comprises six dimeric [Zn5{(OMe)3Br3L}2] units. Whereas 3 and 4 decompose in DMSO solution, 6 is surprisingly stable in the same solvent. PMID- 15786500 TI - Lanthanide complexes coordinated by N-substituted (R)-1,1'-binaphthyl-2,2' diamido ligands in the catalysis of enantioselective intramolecular hydroamination. AB - A new family of lanthanide ionic complexes derived from chiral, substituted (R) binaphthylamine ligands, [Li(thf)(4)][Ln{(R)-C(20)H(12)(NR)(2)}(2)] (Ln=Yb, Sm, Nd, or Lu), has been synthesized and characterized by X-ray crystal structure analyses. All complexes have been tested as new catalysts for the hydroamination/cyclization of 1-(aminomethyl)-1-allylcyclohexane. Ytterbium complexes proved to be both the most active and the most enantioselective, and the use of the complex [Li(thf)(4)][Yb{(R)-C(20)H(12)(NC(3)H(7))(2)}(2)], bearing isopropyl radicals on the nitrogen atoms, allowed the formation of the corresponding spiropyrrolidine in high yield with up to 70 % ee. PMID- 15786502 TI - Diphenylamino end-capped oligofluorenes with enhanced functional properties for blue light emission: synthesis and structure-property relationships. AB - A novel series of monodisperse asymmetrically and symmetrically substituted diphenylamino end-capped oligofluorenes, OF(2)-NPhR, R = H or An (An = 9-anthryl) and OF(n)-NPh, n = 2-4, has been synthesized by a convergent approach using palladium-catalyzed Suzuki cross-coupling. End-capping of oligofluorenes with diphenylamino group(s) has been shown to offer advantages in terms of lowering their first ionization potentials, enhancing thermal stability, and inducing good amorphous morphological stability. By tuning the number of diphenylamino end-caps and the chain length, the optimal conjugated length for optical and luminescence properties has been determined. Of all the hitherto reported oligofluorenes capable of serving as non-doped blue emitters, OF(3)-NPh, with an optimal conjugated length, exhibits some of the best hole-transport and blue-emitting properties. A maximum luminance of 7500 cd m(-2) and a luminance efficiency up to 1.8 cd A(-1) have been achieved. PMID- 15786503 TI - Significant enhancement of the Stille reaction with a new combination of reagents copper(I) iodide with cesium fluoride. AB - The combination of copper(I) iodide and cesium fluoride significantly enhances the Stille reaction. After extensive optimisation, a variety of electronically unfavourable and sterically hindered substrates were coupled in very high yields under mild conditions. PMID- 15786504 TI - Urea and thiourea derivatives as low molecular-mass organogelators. AB - The properties of a series of organogels consisting of a urea or thiourea derivative with one or two n-alkyl substitutuents at the nitrogen atoms (a low molecular-mass organogelator (LMOG)) and an organic liquid are described. They include N,N'-dimethylurea, the LMOG of lowest molecular mass (M(W) 88) we are aware of. The efficiencies of the LMOGs, based the diversity of liquids gelated, the minimum amount required for gelation of a liquid at room temperature, and the temporal and thermal stabilities of the gels formed, have been investigated as a function of the number, length, and substitution pattern of their n-alkyl chains. The gels are thermally reversible and require generally very low concentrations (<2 wt %) of an LMOG. Some of the LMOGs with shorter chains are more efficient than their longer chained analogues. The structural and thermodynamic properties of the gels have been examined by IR, DSC, and X-ray diffraction techniques. Polarizing optical microscopic analyses of the gels show that the nature of gelator aggregates depends mainly on the alkyl chain length. Changes in the aggregation ability have been examined systematically by perturbing the molecular structure. PMID- 15786505 TI - Oxygen atom transfer in models for molybdenum enzymes: isolation and structural, spectroscopic, and computational studies of intermediates in oxygen atom transfer from molybdenum(VI) to phosphorus(III). AB - Intermediates in the oxygen atom transfer from Mo(VI) to P(III), [Tp(iPr)MoOX(OPR3)] (Tp(iPr) = hydrotris(3-isopropylpyrazol-1-yl)borate; X = Cl-, phenolates, thiolates), have been isolated from the reactions of [Tp(iPr)MoO2X] with phosphines (PEt3, PMePh2, PPh3). The green, diamagnetic oxomolybdenum(IV) complexes possess local C(1) symmetry (by NMR spectroscopy) and exhibit IR bands assigned to nu(Mo==O) (approximately 950 cm(-1)) and nu(P==O) (1140-1083 cm(-1)) vibrations. The X-ray crystal structures of [Tp(iPr)MoOX(OPEt3)] (X = OC6H4-2 sBu, SnBu), [Tp(iPr)MoO(OPh)(OPMePh2)], and [Tp(iPr)MoOCl(OPPh3)] have been determined. The monomeric complexes exhibit distorted octahedral geometries, with coordination spheres composed of tridentate fac-Tp(iPr) and mutually cis monodentate terminal oxo, phosphoryl (phosphine oxide), and monoanionic X ligands. The electronic structures and stabilities of the complexes have been probed by computational methods, with the three-dimensional energy surfaces confirming the existence of a low-energy steric pocket that restricts the conformational freedom of the phosphoryl ligand and inhibits complete oxygen atom transfer. The reactivity of the complexes is also briefly described. PMID- 15786506 TI - Stereocontrol in alkyne cyclocarbonylation reactions promoted by a bioxazoline palladium(II) complex. AB - Insertion of 1,2-disubstituted alkynes into [Pd(CH3)(CO)(BIOX)]+[B{3,5 (CF3)2C6H3}4]- (1), where BIOX=(4S,4'S)-(-)-4,4',5,5'-tetrahydro-4,4'-bis(1 methylethyl)-2,2'-bioxazole, leads to the formation of five-membered palladacycles, which, by reaction with carbon monoxide, produce a mixture of two diastereomeric forms of a palladium complex containing an eta3-allylic gamma lactone ligand. On leaving the mixture in solution, one isomer was converted into the other, reaching a diastereomeric excess of up to 94 %. The steric and electronic factors responsible for the epimerization process were investigated by theoretical methods. Cleavage of the eta3-allyl--palladium bond by nucleophiles allowed highly substituted chiral butenolides to be synthesized in good enantiomeric excess. PMID- 15786507 TI - Revealing structural effects, part II: the influence of molecular structure on the adsorption of butanol isomers on platinum. AB - The nature of the adsorbates formed when butanol isomers interact with platinum electrodes in a perchloric acid medium was investigated by the application of on line differential electrochemical mass spectrometry (DEMS) and cyclic voltammetry. In this way, the reactivity of the residues remaining on the electrode surface after a flow-cell experiment was established for the different molecules. It was found that the four isomers form strongly adsorbed species on the electrode, which undergo both electro-oxidation and electroreduction, depending on the potential applied at the electrode. Oxidative stripping of the adsorbates produces CO2 as the only oxidation product, whereas propane and the corresponding butane isomer are obtained on platinum in the hydrogen adsorption potential region. The yields of these hydrocarbons were found to depend strongly on the nature of the butanol isomer and on the adsorption potential. According to these results, it can be concluded that fragmentation of the butanol isomers occurs during adsorption and reduction reactions. C4 alkene and acetyl species are proposed as the adsorbed intermediates in all cases. PMID- 15786508 TI - Post-reperfusion changes of monocyte function in coronary blood after extracorporeal circulation. AB - BACKGROUND: Neutrophil and mononuclear cell functional changes represent a hallmark of inflammation during cardiopulmonary bypass and cardiovascular surgery. Knowledge of mechanisms underlying monocyte functional modulation in coronary blood may be useful to develop protective interventions that can limit ischemia/reperfusion injury. METHODS: Samples of 13 patients were drawn from venous coronary sinus before cardioplegic arrest and after reperfusion. The following parameters were studied: surface molecules expression (CD18, CD11b, CD44, CD162, CD15s, CD80, CD86, CD16, CD49d, CD29, CD25, HLA-DR, Toll-like receptor-4 [TLR-4], CXCR1, CCR2, CCR5, CX3CR1), oxidative burst response, monocyte-platelet conjugates (using antibodies against CD45, CD14, CD41a), and platelet activation (CD62P, PAC-1). Enzyme-linked immunosorbent assays were performed to measure levels of interleukin (IL)-1beta, IL-6, IL-8, IL-10, and tumor necrosis factor-alpha (TNF-alpha). RESULTS: Coronary reperfusion down modulated monocyte molecules expression, especially for CD18 (P = 0.048), CD44 (P = 0.0035), CD49d (P = 0.0029), CD29 (P = 0.032), HLA-DR (P < 0.0001), TLR-4 (P = 0.0109), CCR2 (P = 0.0184), CCR5 (P = 0.0396), and CX3CR1 (P < 0.0001). A marginal increase (P = 0.062) of a normalized adhesion index between monocytes and platelets was observed at reperfusion. No variations were detected for the monocyte oxidative burst and platelet activation. Increased levels of IL-6 (P = 0.013), TNF-alpha (P = 0.0272), and IL-10 (P = 0.0008) were measured after cardioplegia. CONCLUSIONS: The lack of CD11b and CD25 variations and of the oxidative burst exclude monocyte activation at reperfusion. The high after cardioplegia level of IL-10, the decreased expression of HLA-DR and TLR-4, and the absence of IL-1beta and IL-8 suggest an IL-10-mediated functional depression of monocyte, including their adhesive and migratory capacities. The lack of an after-cardioplegia orientation toward IL-10 producing a "macrophage-like" CD14+/CD16+ phenotype might mean that myocardial infiltrating lymphocytes are the main source of IL-10. Moreover, the increased after-cardioplegia levels of IL-6 and TNF-alpha might be due to myocardial and endothelial activations. The increased adhesion index between monocyte and platelets, without receptor variations, suggests a monocyte membrane modification induced by extracorporeal circulation. PMID- 15786509 TI - T-cell lymphoblastic leukemia/lymphoma syndrome with eosinophilia and acute myeloid leukemia. AB - This case represents an example of an unusual T-cell lymphoblastic leukemia/lymphoma syndrome associated with eosinophilia and myeloid malignancy in a young boy. This case is one of only five reported "leukemic" variants of the disease and demonstrates the importance of considering this poor prognostic diagnosis in pediatric acute lymphoblastic leukemia. This case also illustrates the importance of an interactive multidisciplinary approach to the laboratory evaluation of a leukemia patient. PMID- 15786510 TI - Slit-flow ektacytometry: laser diffraction in a slit rheometer. AB - BACKGROUND: Deformability of red blood cells (RBCs) is a determinant of blood flow resistance as RBCs pass through small capillaries of the microcirculation. Available techniques for measuring RBC deformability often require a washing process after each measurement, which is not optimal for day-to-day clinical use. METHODS: A laser diffraction technique has been combined with slit-flow rheometry, which shows significant advances in ektacytometric design, operation, and data analysis. The essential features of this design are its simplicity (ease of operation and no moving parts) and a disposable element that is in contact with the blood sample. RESULTS: With slit ektacytometry, the deformation of RBCs subjected to continuously decreasing shear stress in a slit flow can be quickly measured with extremely small quantities of blood. The measurements with the slit ektacytometer were compared with those of LORCA and a strong correlation was apparent. The deformability of the hardened RBCs was markedly lower than that of the normal RBCs. In addition, the young cells showed higher values of the elongation index than did the old cells. CONCLUSIONS: The newly developed slit ektacytometer can measure RBC deformability with ease and accuracy. In addition, the slit ektacytometer can be easily used in a clinical setting owing to the incorporation of a disposable element that holds the blood sample. PMID- 15786511 TI - Osx transcriptional regulation is mediated by additional pathways to BMP2/Smad signaling. AB - Bone morphogenetic protein (BMP)-2 induces Osterix (Osx) in mouse C2C12 cells and chondrocytes. Genetic studies place Osx downstream to the BMP-2/Smad/Runx2 signaling pathway; however, limited information is available on the mediators of Osx expression in osteoblast lineage commitment. Several lines of research implicate the presence of Runx2-independent ossification. Therefore, the purpose of this study was to identify possible mediators of Osx expression beyond the BMP 2/Smad pathway. Using real-time RT-PCR, we showed upregulation of Osx in response to BMP-2 in human mesenchymal stem cells (hMSC). Insulin-like growth factor (IGF) I upregulated Osx, but not Runx2. Further, IGF-I in combination with BMP-2 was synergistic for Osx, suggesting a pathway beyond Smad signaling. MAPK was tested as a common mediator across BMP-2 and IGF-I signaling pathways. Inhibition of MAPK component ERK1/2 did not affect Runx2 gene expression, but inhibited Osx expression and matrix mineralization. BMP-2-mediated Osx expression was downregulated in response to p38 inhibition. We therefore conclude that during osteogenic lineage progression, in addition to the BMP-2/Smad pathway, IGF-I and MAPK signaling may mediate Osx. PMID- 15786512 TI - Small G-protein Rho is involved in the maintenance of cardiac myocyte morphology. AB - The use of small membrane-permeable sequences or protein transduction domains (PTDs) can facilitate the transport of proteins into many cell types. In preliminary studies with the application of three PTDs (penetratin, modified penetratin, and the HIV TAT transduction domains) to cardiac myocytes, we found that the TAT and penetratin sequences showed high efficiency of uptake and low toxicity. Rho has been previously shown to be an important regulator of cytoskeletal organization and morphology in other non-cardiac cell types. To evaluate a role for Rho in cardiac myocyte morphology, we used the TAT-PTD to deliver a RhoA-specific inhibitor, the C3 exoenzyme, to cultured cardiac myocytes. We showed that this incubation with TAT-C3 abolished the basal levels of RhoA activity, demonstrating the efficacy of this treatment. Incubation with TAT-C3 also altered cardiac myocyte morphology so that TAT-C3-treated cells produced multiple projections from the major cell body. This was accompanied by a statistically significant increase in cell size, albeit to a lesser extent than the changes accompanying exposure to the hypertrophic agent, endothelin-1. Furthermore, the change in size of TAT-C3-treated cells was not accompanied by the induction of atrial natriuretic factor (ANF) expression that accompanies the hypertrophy of cardiac myocytes. These results reveal a role for RhoA in the maintenance of normal myocyte morphology. PMID- 15786513 TI - Exploring the risk of diabetes mellitus and dyslipidemia among ambulatory users of atypical antipsychotics: a population-based comparison of risperidone and olanzapine. AB - PURPOSE: To compare the incidence rates of diabetes mellitus and dyslipidemia in ambulatory first-time users of risperidone and olanzapine. METHODS: The database for the Prescription Drug Insurance Plan in the province of Quebec was used as the data source for a population-based cohort study. Denominalized data were extracted for all ambulatory patients who first received an atypical antipsychotic between 1 January 1997 and 31 August 1999. Eligible patients were categorized as taking: no antidiabetic medication; no lipid reducing medication; neither type of medication. Those who started to use an outcome drug (an antidiabetic or lipid-lowering medication) before the end of the follow-up period (31 August 2000) were considered to have developed the corresponding outcome disease. Incidence rate ratios (IRR) (and 95% confidence intervals) for initiating antihyperglycemic or lipid-lowering drug treatment, or both were calculated. Outcomes on risperidone were compared to those on olanzapine. RESULTS: A total of 19 582 eligible patients were included in the analysis. Relative to risperidone, olanzapine was associated with a higher risk of initiating a pharmacologic treatment for diabetes [IRR: 1.33 (1.03-1.74)], dyslipidemia [IRR: 1.49 (1.22-1.83)], or either condition [1.47 (1.23-1.76)]. CONCLUSIONS: Olanzapine seems to be associated with a higher risk of developing diabetes and/or dyslipidemia than risperidone. Further prospective studies are needed to rigorously assess the safety of olanzapine. PMID- 15786514 TI - The psychological impact of mammographic screening. A systematic review. AB - Concerns have been raised regarding the possible negative psychological impact of the cancer screening programmes offered in the UK. This review aims to assess the extent of and factors associated with the adverse psychological consequences of mammographic screening. Fifty-four papers from 13 countries were identified, a majority of which were published after 1990, coinciding more or less with the onset of routine mammographic screening. The results report that mammographic screening does not appear to create anxiety in women who are given a clear result after a mammogram and are subsequently placed on routine recall. However, women who have further investigations following their routine mammogram experience significant anxiety in the short term, and possibly in the long term. The nature and extent of the further investigation that women are exposed to during mammographic screening determines the intensity of the psychological impact. Factors associated with the adverse psychological impact of mammographic screening included: social demographic factors of younger age, lower education, living in urban areas, manual occupation, and one or no children; cancer screening factors of dissatisfaction with information and communication during screening process, waiting time between recall letter and recall appointment, pain experienced during the mammographic screening procedures, and previous false positive result; and cancer worry factors including fear of cancer and greater perceived risk of breast cancer. Difficulties in measuring the psychological impact of screening are discussed, and methods of alleviating the negative psychological outcomes are suggested. PMID- 15786515 TI - Drug utilization pattern of Chinese herbal medicines in a general hospital in Taiwan. AB - PURPOSE: Drug utilization studies are important for the optimization of drug therapy and have received a great attention in recent years. Most of the information on drug use patterns has been derived from studies in modern Western medicines; however, studies regarding the drug utilization of traditional Chinese medicine (CM) are few. The present study was the first clinical research to evaluate the drug utilization patterns of Chinese herbal medicines in a general hospital in Taiwan. METHODS: Data were collected prospectively from the patients attending the Traditional Medicine Center of Taipei Veteran General Hospital under CM drug treatments. The study was carried out over a period of 1 year, from January 2002 to December 2002. Core drug use indicators, such as the average number of drugs per prescriptions, the dosing frequency of prescriptions, and the most common prescribed CM herbs and formulae were evaluated. The primary diagnosis and the CM drugs prescribed for were also revealed. All data were analyzed by descriptive statistics. RESULTS: A total of 10 737 patients, representing 52 255 CM drugs, were screened during the study period. Regarding the prescriptions, the average number of drugs per prescription was 4.87 and 37.21% of prescriptions were composed by five drugs. Most of prescriptions (91.38%) were prescribed for three times a day. The most often prescribed Chinese herb was Hong-Hwa (5.76%) and the most common Chinese herbal formula was Jia-Wey Shiau-Yau-San (3.80%). The most frequent main diagnosis was insomnia (15.58%), followed by menopause (5.22%) and constipation (5.09%). CONCLUSION: The survey revealed the drug use pattern of CMs in a general hospital. The majority of CM prescriptions were composed by 3-6 drugs and often prescribed for three times a day. Generally, the rational drug uses of CM drugs were provided with respect to the various diagnoses. PMID- 15786516 TI - Antipsychotic exposure and type 2 diabetes among patients with schizophrenia: a matched case-control study of California Medicaid claims. AB - PURPOSE: To examine the risk of developing type 2 diabetes mellitus among people with schizophrenia exposed to atypical antipsychotics (clozapine, olanzapine, quetiapine, risperidone) compared to those exposed to conventional antipsychotics. METHODS: A matched case-control design was used to examine California Medicaid beneficiaries. Cases developed diabetes subsequent to being diagnosed with schizophrenia (ICD-9295), were 18 years or older, and were exposed to at least one antipsychotic medication at some point during the 12 weeks preceding diabetes diagnosis. Diabetes was defined by diagnostic claim (ICD-9250) or prescription for antidiabetic agents. A total of 3663 cases were matched to 14 523 non-diabetic controls (people with schizophrenia matched on gender and age +/ 5 years). All had to be continuously eligible for benefits during the 12-week period preceding diabetes onset in the case. Conditional logistic regression modeled the risk of exposure, controlling for age, ethnicity, and exposure to selected concomitant medications. Analyses were repeated with 24- and 52-week exposure windows. RESULTS: Using a 12-week exposure window, olanzapine (OR = 1.36, 95%CI 1.20-1.53), clozapine (OR = 1.34, 95%CI 1.16-1.55), and combination atypical therapy (OR = 1.58, 95%CI 1.33-1.88), but not risperidone or quetiapine, were associated with increased odds of developing diabetes compared to conventional antipsychotics. Changing to a 24-week exposure window, the risks were: olanzapine (OR = 1.38, 95%CI 1.22-1.56), clozapine (OR = 1.32, 95%CI 1.14 1.53), or combinations (OR = 1.54, 95%CI 1.29-1.84). With a 52-week exposure window, the risks were: olanzapine (OR = 1.41, 95%CI 1.24-1.60), clozapine (OR = 1.41, 95%CI 1.21-1.65), combinations (OR = 1.58, 95%CI 1.31-1.90). Risk for olanzapine increased with dose. Hispanic, African American, and unknown ethnicity were significant risks for development of type 2 diabetes as was exposure to selected concomitant medications. CONCLUSIONS: Exposure to olanzapine or clozapine is associated with a 34-41% increase in the developing of type 2 diabetes among California Medicaid recipients with schizophrenia. Prospective, randomized trials are needed to confirm these retrospective, observational findings. PMID- 15786517 TI - Comparative effects of glucose-lysine versus glucose-methionine Maillard reaction products consumption: in vitro and in vivo calcium availability. AB - The influence of glucose-lysine and glucose-methionine Maillard reaction products (MRPs) on calcium availability was studied in rats and in Caco-2 cells. Equimolar glucose/lysine and glucose/methionine mixtures (40% moisture) were heated (150 degrees C, 30 or 90 min) to prepare samples (GL30, GL90, GM30, and GM90, respectively). For 21 days, the rats were fed the AIN-93G diet (control group) or diets containing separately 3% of the heated mixtures (GL30, GL90, GM30, and GM90 groups, respectively). In the last week of the trial, a calcium balance was performed. On day 21, the animals were sacrificed and their livers and femurs removed for analysis of calcium levels. The GL30 and GM30 samples and the corresponding raw mixtures were used for Caco-2 cells experiments. Fecal excretion of calcium decreased and urinary elimination increased in the GM30 and GM90 groups. In accordance, increased calcium transport in Caco-2 cells was found in the presence of the GM30 sample, compared with the raw sample. Bone calcium concentration was lower among the animals consuming MRP diets, compared with the control group. The possible long-term effects of MRP intake on calcium deposition in the bone should be further studied to ascertain the implications on related diseases. PMID- 15786518 TI - Resveratrol and breast cancer chemoprevention: molecular mechanisms. AB - Despite years of intensive research, breast cancer remains a major cause of death among women. New strategies to combat breast cancer are being developed, one of the most exciting of which is the use of chemopreventive agents. Resveratrol (RES) is a polyphenolic compound found in plants that seems to have a wide spectrum of biological activity. RES has been shown to afford protection against several types of cancer. This review summarizes the chemopreventive effects of RES at the three major stages of breast carcinogenesis: initiation, promotion, and progression. It has anti-oxidant and anti-inflammatory properties, and may induce apoptosis as well as modulate cell cycle and estrogen receptor function in breast cancer cell lines. Although RES has shown remarkable promise as a potent chemopreventive agent in breast cancer, further studies are needed to etablish its usefulness. PMID- 15786519 TI - A trinuclear Eu(III) array within a diastereoselectively self-assembled helix formed by chiral bipyridine-carboxylate ligands. PMID- 15786520 TI - Molecular transporter between polymer platforms: highly efficient chemoenzymatic glycopeptide synthesis by the combined use of solid-phase and water-soluble polymer supports. PMID- 15786521 TI - Structural studies of the system Na(saccharinate)n H2O: a model for crystallization. PMID- 15786523 TI - Circular dichroism of a chiral tethered donor-acceptor system: enhanced anisotropy factors in charge-transfer transitions by dimer formation and by confinement. PMID- 15786522 TI - A microfluidic approach for screening submicroliter volumes against multiple reagents by using preformed arrays of nanoliter plugs in a three-phase liquid/liquid/gas flow. PMID- 15786524 TI - Alternating copolymerization of ethylene and butadiene with a neodymocene catalyst. PMID- 15786526 TI - Randomized trial comparing the Prolene Hernia System, mesh plug repair and Lichtenstein method for open inguinal hernia repair (Br J Surg 2005; 92: 33-38). PMID- 15786525 TI - Direct detection of inter-residue hydrogen bonds in polysaccharides by single molecule force spectroscopy. PMID- 15786528 TI - Laparoscopic restorative proctocolectomy (Br J Surg 2005; 92: 88-93). PMID- 15786529 TI - Effects of folic acid on epithelial apoptosis and expression of Bcl-2 and p53 in premalignant gastric lesions. AB - AIM: To evaluate the effects of folic acid on epithelial apoptosis and expression of Bcl-2 and p53 in the tissues of premalignant gastric lesions. METHODS: Thirty eight patients, with premalignant gastric lesions including 18 colonic-type intestinal metaplasia (IM) and 20 mild or moderate dysplasia, were randomly divided into a treatment group (n = 19) receiving folic acid 10 mg thrice daily and a control group (n = 19) receiving sucralfate 1,000 mg thrice daily for 3 mo. All patients underwent endoscopies and four biopsies were taken prior to treatment and repeated after concluding therapy. Folate concentrations in gastric mucosa were measured with chemiluminescent enzyme immunoassay. Epithelial apoptosis and the expression of Bcl-2 and p53 protein in gastric mucosa were detected with flow cytometric assay. RESULTS: The mean of folate concentration in gastric mucosa was 9.03+/-3.37 microg/g wet wt in the folic acid treatment group, which was significantly higher than 6.83+/-3.02 microg/g wet wt in the control group. Both the epithelial apoptosis rate and the tumor suppressor p53 expression in gastric mucosa significantly increased after folic acid treatment. In contrast, the expression of Bcl-2 oncogene protein decreased after folic acid therapy. CONCLUSION: These data indicate that folic acid may play an important role in the chemoprevention of gastric carcinogenesis by enhancing gastric epithelial apoptosis in the patients with premalignant lesions. PMID- 15786530 TI - Effects of 7.5% hypertonic saline on fluid balance after radical surgery for gastrointestinal carcinoma. AB - AIM: To investigate the effects of 7.5% hypertonic saline on positive fluid balance and negative fluid balance, after radical surgery for gastrointestinal carcinoma. METHODS: Fifty-two patients with gastrointestinal carcinoma undergoing radical surgery were studied. The patients were assigned to receive either Ringer lactate solution following 4 mL/kg of 7.5% hypertonic saline (the experimental group, n = 26) or Ringer lactate solution (the control group, n = 26) during the early postoperative period in SICU. Fluid infusion volumes, urine outputs, fluid balance, body weight change, PaO2/FiO2 ratio, anal exhaust time as well as the incidence of complication and mortality were compared between the two groups. RESULTS: Urine outputs on the operative day and the first postoperative day in experimental group were significantly more than in control group (P<0.000001, P = 0.000114). Fluid infusion volumes on the operative day and the first postoperative day were significantly less in experimental group than in control group (P = 0.000042, P = 0.000415). The volumes of the positive fluid balance on the operative day and during the first 48 h after surgery, in experimental group, were significantly less than in control group (P<0.000001). Body weight gain post surgery was significantly lower in experimental group than in control group (P<0.000001). The body weight fall in experimental group occurred earlier than in control group (P<0.000001). PaO2/FiO2 ratio after surgery was higher in experimental group than in control group (P = 0.000111). The postoperative anal exhaust time in experimental group was earlier than in control group (P = 0.000006). The overall incidence of complications and the incidence of pulmonary infection were lower in experimental group than in control group (P = 0.0175, P = 0.0374). CONCLUSION: 7.5% hypertonic saline has an intense diuretic effect and causes mobilization of the retained fluid, which could reduce fluid infusion volumes and positive fluid balance after radical surgery for gastrointestinal carcinoma, as well as, accelerate the early appearance of negative fluid balance after the surgery, improve the oxygen diffusing capacity of the patients' alveoli, and lower the overall incidence of complications and pulmonary infection after the surgery. PMID- 15786531 TI - Nutritional status and quality of life of the gastric cancer patients in Changle County of China. AB - AIM: To analyze the relation between nutrition and quality of life in the stomach cancer patients, evaluate the intake of daily nutrition of the patients, and study the feasibility of nutrition intervention in improving quality of life of the stomach cancer patients. METHODS: A total of 285 surgical stomach cancer patients reported in the Changle Cancer Registry from 2002 to 2003 were investigated with respect to their diet and quality of life. Daily nutrition intakes of the patients were calculated according to the Food Composition Database, and these data were compared with the reference values proposed by the Chinese Nutrition Society. The partial correlation was used to analyze the relationship between nutrition and quality of life in the patients. Stepwise multiple regression analyses were conducted to analyze the factors influencing nutrition intake in stomach cancer patients. RESULTS: Except vitamin C, there were statistical correlations between the nutrition and quality of life in stomach cancer patients, and differences of the daily nutrition intake among three groups (good, modest and bad quality of life) of the patients were significant. Most of the stomach cancer patients had a lower daily nutrition intake than the reference values. At the significance level alpha = 0.05, the factors influencing the daily nutrition intake of the patients were number of meals a day, family income, way of operation, exercise and age. CONCLUSION: The nutritional status of the operated patients with stomach cancer may impact on their quality of life. The stomach cancer patients in Changle County have a low level of daily nutrition intake, which suggests that they have a bad nutritional status. To improve the quality of life of the patients, the nutrition intervention should be conducted. Increasing times of meals a day and having a high-protein, high-calorie foods can improve the nutritional status of the stomach cancer patients. Moreover, exercise for rehabilitation can whet the appetite of the patients and recover their body function, which in turn may improve the quality of life of the stomach cancer patients. PMID- 15786532 TI - Feasible economic strategies to improve screening compliance for colorectal cancer in Korea. AB - AIM: While colorectal cancer (CRC) is an ideal target for population screening, physician and patient attitudes contribute to low levels of screening uptake. This study was carried out to find feasible economic strategies to improve the CRC screening compliance in Korea. METHODS: The natural history of a simulated cohort of 50-year-old Korean in the general population was modeled with CRC screening until the age of 80 years. Cases of positive results were worked up with colonoscopy. After polypectomy, colonoscopy was repeated every 3 years. Baseline screening compliance without insurance coverage by the national health insurance (NHI) was assumed to be 30%. If NHI covered the CRC screening or the reimbursement of screening to physicians increased, the compliance was assumed to increase. We evaluated 16 different CRC screening strategies based on Markov model. RESULTS: When the NHI did not cover the screening and compliance was 30%, non-dominated strategies were colonoscopy every 5 years (COL5) and colonoscopy every 3 years (COL3). In all scenarios of various compliance rates with raised coverage of the NHI and increased reimbursement of colonoscopy, COL10, COL5 and COL3 were non-dominated strategies, and COL10 had lower or minimal incremental medical cost and financial burden on the NHI than the strategy of no screening. These results were stable with sensitivity analyses. CONCLUSION: Economic strategies for promoting screening compliance can be accompanied by expanding insurance coverage by the NHI and by increasing reimbursement for CRC screening to providers. COL10 was a cost-effective and cost saving screening strategy for CRC in Korea. PMID- 15786534 TI - Evaluation of diffusion in gel entrapment cell culture within hollow fibers. AB - AIM: To investigate diffusion in mammalian cell culture by gel entrapment within hollow fibers. METHODS: Freshly isolated rat hepatocytes or human oral epidermoid carcinoma (KB) cells were entrapped in type I collagen solutions and statically cultured inside microporous and ultrafiltration hollow fibers. During the culture time collagen gel contraction, cell viability and specific function were assessed. Effective diffusion coefficients of glucose in cell-matrix gels were determined by lag time analysis in a diffusion cell. RESULTS: Significant gel contractions occurred in the collagen gels by entrapment of either viable hepatocytes or KB cells. And the gel contraction caused a significant reduction on effective diffusion coefficient of glucose. The cell viability assay of both hepatocytes and KB cells statically cultured in hollow fibers by collagen entrapment further confirmed the existence of the inhibited mass transfer by diffusion. Urea was secreted about 50% more by hepatocytes entrapped in hollow fibers with pore size of 0.1 mum than that in hollow fibers with MWCO of 100 ku. CONCLUSION: Cell-matrix gel and membrane pore size are the two factors relevant to the limited mass transfer by diffusion in such gel entrapment of mammalian cell culture. PMID- 15786533 TI - Association of polymorphisms of interleukin-18 gene promoter region with chronic hepatitis B in Chinese Han population. AB - AIM: To investigate the polymorphisms of interleukin-18 (IL-18) gene promoters, and to disclose whether such polymorphisms are associated with susceptibility to chronic hepatitis B in Chinese Han population. METHODS: Using polymerase chain reaction with sequence specific primers (PCR-SSP) method, the single nucleotide polymorphisms (SNPs) of the promoter region of IL-18 gene at position -607 and 137 were detected in 231 patients with chronic hepatitis B and 300 normal controls. RESULTS: Allele C at position -607 in the promoter of IL-18 gene was detected in 48.7% of normal controls and 51.9% of patients, while allele A at position -607 was detected in 51.3% of normal controls and 48.1% of patients. The frequencies of -607CC, -607 CA and -607AA genotypes in normal controls were 22.0%, 53.3% and 24.7% respectively and in chronic hepatitis B patients were 26.8%, 50.2% and 23.0% respectively. Allele G at position -137 in the promoter of IL-18 gene was detected in 82.3% of normal controls and 88.5% of chronic hepatitis B patients, while allele C at position -137 was detected in 17.7% of normal controls and 11.5% of patients. The frequencies of -137GG, GC and CC genotype were 67.3%, 30.0% and 2.7% in normal controls respectively, while in chronic hepatitis B patients were 78.8%, 19.5% and 1.7% respectively. The frequency of -137GG genotype in chronic hepatitis B groups was significantly higher than that in normal controls (chi2 = 8.55, P = 0.003<0.05), whereas the frequencies of -607C/-137C and -607A/-137C haplotypes in chronic hepatitis B groups were significantly lower than that in normal controls. The association between genotypes of IL-18 promoter region polymorphisms and HBV copies showed that the frequency of -607AA genotype in high HBV-DNA copies groups was lower than that in low HBV-DNA copies groups (chi2 = 6.03, P = 0.014<0.05). CONCLUSION: The polymorphisms of the promoter region of IL-18 gene at position -607 and -137 are closely associated with susceptibility to chronic hepatitis B. The people with allele C at position -137 in the promoter of IL-18 gene may be protected against HBV infection; moreover AA genotype at position -607 may be closely linked to inhibit HBV-DNA replication. These findings give some new clues to the study of pathogenesis of chronic hepatitis B. PMID- 15786535 TI - Attenuation of graft ischemia-reperfusion injury by urinary trypsin inhibitor in mouse intestinal transplantation. AB - AIM: Ischemia/reperfusion (I/R) injury is one of the major obstacles for intestinal transplantation (ITx). Urinary trypsin inhibitor (Ulinastatin, UTI) suppresses proteases and stabilizes lysosomal membranes. We supposed that Ulinastatin would diminish I/R injury of intestinal graft. METHODS: UTI- treated group and untreated control group were investigated by histological assessment at 1.5, 4, 24, and 72 h after ITx. Myeloperoxidase (MPO) activity was used as the activity of neutrophils, and malondialdehyde (MDA) was used as an index of lipid peroxidation. TNFalpha and i-NOS mRNA expression in graft tissue were measured by semi-quantitative RT-PCR. CD11b+Gr1+ cells in graft lamina propria were analyzed by flow cytometry. RESULTS: Histological scores of the graft showed that the tissue injury was markedly attenuated by UTI treatment at different time points after ITx, with reduced MPO and MDA value in the grafts. The expression of TNFalpha and i-NOS mRNA was profoundly inhibited, while the infiltration of CD11b+ Gr1+ cells into the intestinal graft was decreased in UTI group. CONCLUSION: Urinary trypsin inhibitor attenuates I/R injury in mouse intestinal transplantation by reducing monocytes infiltration and down-regulation of TNFalpha and i-NOS mRNA expression. PMID- 15786536 TI - Analysis of the mechanisms of rabbit's brainstem hemorrhage complicated with irritable changes in the alvine mucous membrane. AB - AIM: To explore the dynamic changes in the pressure of the lateral ventricle during acute brainstem hemorrhage and the changes of neural discharge of vagus nerve under the load of intracranial hypertension, so as to analyze their effects on the congestive degree of intestinal mucous membrane and the morphologic changes of intestinal mucous membrane. METHODS: An operation was made to open the skull to obtain an acute brainstem hemorrhage animal model. Microcirculatory microscope photography device and video recording system were used to determine the changes continuously in the caliber of jejunal mesenteric artery during brainstem hemorrhage and the changes with time in the congestion of jejunal mucosal villi. We used HE stain morphology to analyze the changes of duodenal mucosal villi. A recording electrode was used to calculate and measure the electric discharge activities of cervical vagus nerve. RESULTS: (1) We observed that the pressure of lateral cerebral ventricle increased transiently during acute brainstem hemorrhage; (2) The caliber of the jejunal mesenteric artery increased during brainstem hemorrhage. Analysis of red color coordinate values indicated transient increase in the congestion of jejunal mucous membrane during acute brainstem hemorrhage; (3) Through the analysis of the pathologic slice, we found enlarged blood vessels, stagnant blood, and transudatory red blood cells in the duodenal submucous layer; (4) Electric discharge of vagus nerve increased and sporadic hemorrhage spots occurred in duodenal mucous and submucous layer, when the lateral ventricle was under pressure. CONCLUSION: Brainstem hemorrhage could cause intracranial hypertension, which would increase the neural discharge of vagus nerve and cause the transient congestion of jejunal mucous membrane. It could cause hyperemia and diffused hemorrhage in the duodenal submucous layer 48 h after brainstem hemorrhage. PMID- 15786537 TI - A community-based epidemiological study of elevated serum alanine aminotransferase levels in Kinmen, Taiwan. AB - AIM: To explore any gender-related differences in prevalence of and condition associated factors related to an elevated serum alanine aminotransferase (ALT) level amongst residents of Kinmen, Taiwan. METHODS: A total of 11,898 of a potential 20,112 regional residents aged 30 years or more completed a related questionnaire that was carried out by the Yang-Ming Crusade between 1991 and 1994 inclusively, with blood samples being collected by public nurses. The overall questionnaire response rate was 59.3% (52.4% for males and 66.0% for females). RESULTS: The prevalence of an elevated serum ALT level for this sub-population was found to be 7.2%, the prevalence revealing a statistically significant decrease with increasing population age (P<0.0001). Males exhibited a greater prevalence of elevated serum ALT level than did females (9.4% vs 5.3%, P<0.0001). Using multiple logistic regression analysis, in addition to male gender, a younger age, greater waist circumference, presence of type-2 diabetes and hyperuricemia were the significant factors associated with an elevated serum ALT level for both males and females. Gender-related differences as regards associated factors were also revealed. For males, obesity was significantly related to an elevated serum ALT level (OR = 1.28, 95%CI: 1.00-1.66) but this was not so for females (OR = 1.09, 95%CI: 0.84-1.42). Hypertriglyceridemia (OR = 1.80, 95%CI: 1.36-2.39) and hyperuricemia (OR = 1.61, 95%CI: 1.03-2.52) were significantly related to elevated serum ALT levels only for females. CONCLUSION: Several gender-related differences were noted pertaining to the prevalence of and relationship between obesity, hypertriglyceridemia and hyperuricemia and elevated serum ALT level in the present study. PMID- 15786538 TI - Metabolic changes in the lower esophageal sphincter influencing the result of anti-reflux surgical interventions in chronic gastroesophageal reflux disease. AB - AIM: With the availability of a minimally invasive approach, anti-reflux surgery has recently experienced a renaissance as a cost-effective alternative to life long medical treatment in patients with gastroesophageal reflux disease (GERD). We are not aware of the fact whether reflux episodes causing complaints for a long time i.e., at least for one year are associated with metabolic changes in the lower esophageal sphincter, and if so, whether these may influence functional results achieved after anti-reflux surgery. METHODS: Between 1 January 2001 and 31 December 2002 we performed anti-reflux surgery on 79 patients. Muscle samples were taken from the lower esophageal sphincter (LES) in 33 patients during anti reflux surgery. Inclusion criteria were: LES resting pressure below 10 mmHg and a marked, pH proven acid exposure to the esophagus of at least one year's duration, causing subjective complaints and requiring continuous proton pump inhibitor treatment. Control samples were obtained from muscle tissue in the gastroesophageal junction that had been removed from 17 patients undergoing gastric or esophageal resection. Metabolic and lysosomal enzyme activities and special protein concentrations 16 parameters in total were evaluated in tissue taken from control specimens and tissue taken from patients with GERD. The biochemical parameters of these intra-operative biopsies were used to correlate the results of anti-reflux operations (Visick I and II-III). RESULTS: In the reflux-type muscle, we found a significant increase of the energy-enzyme activities e.g., creatine kinase, lactate dehydrogenase, beta-hydroxybutyrate dehydrogenase, and aspartate aminotransaminase-. The concentration of the structural protein S-100 and the myofibrillar protein troponin I were also significantly increased. Among lysosomal enzymes, we found that the activities of cathepsin B, tripeptidyl-peptidase I, dipeptidyl-peptidase II, beta hexosaminidase B, beta-mannosidase and beta-galactosidase were significantly decreased as compared to the control LES muscles. By analyzing the activity values of the 9 patients in Visick groups II and III at two months post-surgery, we found a significant increase in the activity of the so-called energy-enzyme values and in the concentration of structural and myofibrillar proteins as compared to the rest of the reflux patients. CONCLUSION: Our results call attention to the metabolic changes that occurred in the LES muscles of reflux patients. The developing hypertrophy-like changes of LES muscles may be a reason for complaints after anti-reflux surgery, which consisted mainly of reports of persisting dysphagia. PMID- 15786539 TI - Sofalcone, a mucoprotective agent, increases the cure rate of Helicobacter pylori infection when combined with rabeprazole, amoxicillin and clarithromycin. AB - AIM: The mucoprotective agents, sofalcone and polaprezinc have anti-Helicobacter pylori (H pylori) activities. We determined the therapeutic effects of sofalcone and polaprezinc when combined with rabeprazole, amoxicillin and clarithromycin for Helicobacter pylori infection. METHODS: One hundred and sixty-five consecutive outpatients with peptic ulcer and H pylori infection were randomly assigned to one of the following three groups and medicated for 7 d. Group A: triple therapy with rabeprazole (10 mg twice daily), clarithromycin (200 mg twice daily) and amoxicillin (750 mg twice daily). Group B: sofalcone (100 mg thrice daily) plus the triple therapy. Group C: polaprezinc (150 mg twice daily) plus the triple therapy. Eradication was considered successful if (13)C-urea breath test was negative at least 4 wk after cessation of eradication regimens or successive famotidine in the cases of active peptic ulcer. RESULTS: On intention to-treat basis, H pylori cure was achieved in 43 of 55 (78.2%) patients, 47 of 54 (87.0%) and 45 of 56 (80.4%) for the groups A, B and C respectively. Using per protocol analysis, the eradication rates were 81.1% (43/53), 94.0% (47/50) and 84.9% (45/53) respectively. There was a significant difference in the cure rates between group A and B. Adverse events occurred in 10, 12 and 11 patients, from groups A, B and C respectively, but the events were generally mild. CONCLUSION: The addition of sofalcone, but not polaprezinc, significantly increased the cure rate of H pylori infection when combined with the rabeprazole-amoxicillin clarithromycin regimen. PMID- 15786540 TI - A Thai family with hereditary pancreatitis and increased cancer risk due to a mutation in PRSS1 gene. AB - AIM: To investigate mutation of serine protease 1-cationic trypsinogen (CT, PRSS1) gene in members of a Thai family with hereditary pancreatitis and pancreatic cancer. METHODS: Polymerase chain reaction and direct sequencing were performed to analyze the PRSS1 gene in two members of the family affected by pancreatitis. Allele specific amplification (ASA) method was then developed to detect the mutation of the PRSS1 gene in all available members of the family and normal control subjects. RESULTS: A cytosine (C) to thymine (T) mutation at position 2441 (g.2441C>T) of the PRSS1 gene, which results in a substitution of arginine by cysteine at position 116 (R116C) of CT, was identified by direct sequencing in both clinically affected members of the family but was not found in the unaffected member. This mutation, which might be arising from deamination of methylated cytosine in CpG dinucleotide of codon 116 (CGT>TGT), was also detected by the ASA method in the two affected members and a proband's brother but was not observed in unaffected members and 54 normal control subjects. CONCLUSION: Autosomal dominant pancreatitis with increased cancer risk in the studied Thai family is most likely due to missense (R116C) mutation in the PRSS1 gene. PMID- 15786541 TI - Serum IL-6, TNFalpha and CRP levels in Greek colorectal cancer patients: prognostic implications. AB - AIM: The significance of preoperative serum IL-6, TNFalpha and CRP levels in the progression of colorectal cancer (CRC) has not been fully elucidated. Our intention was to investigate their role and identify their prognostic significance. METHODS: The IL-6, TNFalpha and CRP levels were measured in 74 CRC patients and the relationships between their elevations and both the clinicopathological factors and prognosis of patients were investigated. Serum concentrations of human IL-6 and TNFalpha were determined by enzyme-linked immunosorbent assay (ELISA). CRP was measured by an immunoturbinometric method. RESULTS: Median IL-6, TNFalpha and CRP levels were significantly higher in CRC patients than in normal controls. High levels of serum IL-6, TNFalpha and CRP were correlated with larger tumor size. Furthermore, high IL-6 and high CRP levels were associated with reduced overall survival. CONCLUSION: Serum IL-6, TNFalpha and CRP levels definitely increase in CRC patients. Pre-operative serum elevation of IL-6 and CRP was thus found to be predictor of the prognosis of CRC patients. The clinical value of TNFalpha in CRC needs to be further investigated. PMID- 15786542 TI - Impact of Helicobacter pylori infection on ghrelin and various neuroendocrine hormones in plasma. AB - AIM: Ghrelin, an endogenous ligand for growth hormone secretagogue receptor, influences appetite, energy balance, gastric motility and acid secretion. The stomach is the main source of circulating ghrelin. There are inconsistent reports on the influence of Helicobacter pylori (H pylori) infection on circulating ghrelin levels. We sought to elucidate the relationship between ghrelin and various peptides in plasma, with special reference to H pylori. METHODS: Plasma ghrelin levels were measured by radioimmunoassay in 89 subjects who were referred for upper gastrointestinal endoscopy, consisting of 42 H pylori infected and 47 uninfected ones. Plasma gastrin, somatostatin, leptin, insulin-like growth hormone 1 (IGF-1) and chromogranin A concentrations were also measured. Twelve patients were treated with anti- H pylori regimen. RESULTS: Ghrelin circulating levels were greatly decreased in H pylori-positive than negative individuals (194.2+/-90.2 fmol/mL and 250.4+/-84.1 respectively, P<0.05), but did not significantly alter following the cure of infection (176.5+/-79.5 vs 191.3+/ 120.4). There was a significant negative correlation between circulating ghrelin and leptin levels, as well as body mass index, for the whole and uninfected population, but not in H pylori-infected patients. Plasma ghrelin concentrations correlated positively with IGF-1 in H pylori-negative group and negatively with chromogranin A in the infected group. There were no significant correlations among circulating levels of ghrelin, gastrin and somatostatin irrespective of H pylori status. CONCLUSION: H pylori infection influences plasma ghrelin dynamics and its interaction with diverse bioactive peptides involved in energy balance, growth and neuroendocrine function. PMID- 15786543 TI - A prospective cross-over study using a sphincterotome and a guidewire to increase the success rate of common bile duct cannulation. AB - AIM: During endoscopic retrograde cholangiopanc-reatography (ERCP), selective cannulation of the common bile duct (CBD) is required in most of the cases. METHODS: From June 2001 till December 2002, all patients referred to our unit for ERCP were considered for entry into the study. Selective CBD cannulation was first attempted with a standard catheter with or without the use of a guidewire. In cases, where CBD cannulation was considered unsuccessful, patients were crossed over to a double-lumen sphincterotome and a guidewire. All patients were hospitalized for 24 h after the procedure in order to assess the incidence of post-ERCP complications. RESULTS: The study sample consisted of 158 patients. Selective CBD cannulation using a standard ERCP catheter with or without the assistance of a guidewire, was accomplished in 129 patients (success rate: 81.65%). From the 29 patients who were crossed over to a sphincterotome and a guidewire, selective CBD cannulation was achieved in 24; the overall success rate rising to 96.8%. Meanwhile, the use of this technique did not increase the incidence of post-ERCP complications. CONCLUSION: The use of a sphincterotome and a guidewire increases the success rate of selective bile duct cannulation in cases that this has not been accomplished with a standard catheter. PMID- 15786544 TI - Metabolic syndrome as a risk factor for gallstone disease. AB - AIM: To establish an association between the presence of metabolic syndrome and the development of gallstone disease. METHODS: We carried out a cross-sectional study in a check-up unit in a university hospital in Mexico City. We enrolled 245 subjects, comprising 65 subjects with gallstones (36 women, 29 men) and 180 controls (79 women and 101 men without gallstones). Body mass index, waist circumference, blood pressure, plasma insulin, and serum lipids and lipoproteins levels were measured. Insulin resistance was calculated by homeostasis model assessment. Unconditional logistic regression analysis (univariate and multivariate) was used to calculate the risk of gallstone disease associated with the presence of at least three of the criteria (Adult Treatment Panel III). Analyses were adjusted for age and sex. RESULTS: Among 245 subjects, metabolic syndrome was present in 40% of gallstone disease subjects, compared with 17.2% of the controls, adjusted by age and gender (odds ratio (OR) = 2.79; 95%CI, 1.46 5.33; P = 0.002), a dose-dependent effect was observed with each component of metabolic syndrome (OR = 2.36, 95%CI, 0.72-7.71; P = 0.16 with one component and OR = 5.54, 95%CI, 1.35-22.74; P = 0.02 with four components of metabolic syndrome). Homeostasis model assessment was significantly associated with gallstone disease (adjusted OR = 2.25; 95%CI, 1.08-4.69; P = 0.03). CONCLUSION: We conclude that as for cardiovascular disease and diabetes mellitus, gallstone disease appears to be strongly associated with metabolic syndrome. PMID- 15786545 TI - "Defective" mutations of hepatitis D viruses in chronic hepatitis D patients. AB - AIM: To verify whether "defective" mutations existed in hepatitis D virus (HDV). METHODS: Hepatitis delta antigen (HDAg)-coding sequences were amplified using Pfu DNA polymerases with proof-reading activities from sera of five patients with chronic hepatitis D. Multiple colonies were sequenced for each patient. Pfu analyzed a total of 270 HDV clones. Three representative defective HDV clones were constructed in expression plasmids and transfected into a human hepatoma cell line. Cellular proteins were extracted and analyzed by Western blot. RESULTS: Four of five cases (80%) showed defective HDV genomes in their sera. The percentage of defective genomes was 3.7% (10/270). The majority (90%) of the defective mutations were insertions or deletions that resulted in frameshift and abnormal stop translation of the HDAg. The predicted mutated HDAg ranged from 45 amino acids to >214 amino acids in length. Various domains of HDAg associated with viral replication or packaging were affected in different HDV isolates. Western blot analysis showed defected HDAg in predicted positions. CONCLUSION: "Defective" viruses do exist in chronic HDV infected patients, but represented as minor strains. The clinical significance of the "defected" HDV needs further study to evaluate. PMID- 15786546 TI - Ribavirin monotherapy increases sustained response rate in relapsers of end treatment virologic responders. AB - AIM: To assess the efficacy of ribavirin monotherapy in patients with biochemical relapse after combination therapy. METHODS: Twenty-four weeks of ribavirin monotherapy was given to biochemical relapsers of end treatment biochemical responders within 6 mo after combination therapy, including non-responders with HCV-RNA level < or =0.2 Meq/mL and end treatment virologic responders (ETVRs) with or without reappearance of HCV-RNA. RESULTS: Sixty-two chronic HCV-infected patients completed 24 wk of interferon-alpha plus ribavirin combination therapy. Fifty patients (80%) achieved end treatment biochemical response including 16 non responders and 34 of 36 ETVRs. Twenty-six patients (41.9%) were non-responders. Ribavirin monotherapy was given to 20 biochemical relapsers including 12 non responders with HCV-RNA levels < or =0.2 Meq/mL, four of eight HCV-RNA reappearing ETVRs, and four HCV-RNA negative ETVRs. After 24 wk of ribavirin monotherapy, one of 12 non-responders, two of four HCV-RNA reappearing ETVRs and all four RNA-negative biochemical relapsers of ETVRs showed sustained virologic response. Two of 12 monotherapy treated non-responders showed persistent normalization of liver function test. In total, 50% (31/62) of patients achieved sustained virologic response. CONCLUSION: Resumption of ribavirin monotherapy in ETVRs at signs of viral rebound and recurrent biochemical abnormalities rather than continuation of monotherapy appears to be the key to success of ribavirin monotherapy after interferon-related combination therapy. PMID- 15786547 TI - Detailed deletion mapping of loss of heterozygosity on 22q13 in sporadic colorectal cancer. AB - AIM: Both development and progression of malignancies occur as a multistep process, requiring the activation of oncogenes and the inactivation of several tumor suppressor genes. The loss of heterozygosity (LOH) of tumor suppressor genes is believed to play a key role in carcinogenesis of colorectal cancer (CRC). In this study, we analyzed the LOH of seven loci on chromosome 22q13 in an effort to identify candidate tumor suppressor genes involved in colorectal carcinogenesis. METHODS: Matched tumor and normal tissue DNA were analyzed by PCR using fluorescence-labeled polymorphic microsatellite markers in 83 CRC patients. PCR products were eletrophoresed and LOH was determined by calculating the peak height acquired through computer software. Comparisons between LOH frequency and clinicopathological features were performed by chi2 test. P<0.05 was considered as statistical significance. RESULTS: The average LOH frequency of chromosome 22q13 was 28.38%. The highest LOH frequency was 64.71% on D22S1160 locus, and the lowest was 21.43% on D22S1141 locus. We detected two obvious minimal deletion regions: one between markers D22S1171 and D22S274, the other flanked by markers D22S1160 and D22S1149, each about 2.7 and 1.8 cm, respectively. None had lost in all informative loci. LOH frequency on D22S1171 is 50% on distal colon, which was higher than that on proximal one (P = 0.020); on D22S114 locus, none LOH event occurred in patients with liver metastasis, whilst 46.94% occurred in patients without liver metastasis (P = 0.008); on D22S1160 locus, LOH frequency in lymph nodes metastasis patients was 83.33%, which was much higher than 43.75% without lymph nodes metastasis ones (P = 0.016). There was no statistical significance between clinicopathological features and other loci. CONCLUSION: This study provides evidence of two minimal deletion regions, which may harbor putative tumor suppressor genes related to progression and metastasis in sporadic colorectal carcinoma on chromosome 22q13. PMID- 15786548 TI - Clinicopathological and molecular genetic analysis of HNPCC in China. AB - AIM: To explore the clinicopathological and molecular genetic features of hereditary nonpolyposis colorectal cancer (HNPCC) in Chinese population. METHODS: We collected 16 Chinese HNPCC families from Wenzhou, Zhejiang Province, China. Tumor tissues and peripheral white blood cells were studied using microdissection, microsatellite analysis, immunostaining of hMSH2 and hMLH1 proteins and direct DNA sequencing of hMSH2 and hMLH1 genes. RESULTS: (1) A total of 50 patients had CRC. Average age at diagnosis of the first CRC was 45.7 years; 40.9% and 28.7% of the CRCs were located proximal to the splenic flexure and in the rectum, respectively. Thirty-eight percent of the colorectal cancer patients had synchronous and metachronous CRC. 34.4% and 25% of the CRCs were poor differentiation cancer and mucinous adenocarcinoma, respectively. Fourteen extracoloni tumors were found, and the hepatic cancer was the most common tumor type. Twenty-one patients whose median survival time was 5.7 years died during 1 23 years. Twenty-nine patients have survived for 1-28 years, 58.6%, 41.4% and 24.1% patients have survived for more than 5, 10 and 15 years, respectively; (2) All nine tumor-tissues showed microsatellite instability (MSI) at more than two loci. Four tumor-tissues lost hMSH2 protein expression and one lost hMLH1 protein expression. Three pathological germline mutations were identified from five genetically analyzed families; two of three mutations had not been reported previously as they were a transition from C to A in exon 14 (codon 743) of hMSH2 and a TTC deletion in exon 14 (codon 530) of hMLH1. CONCLUSION: Chinese HNPCC have specific clinicopathological features, such as early onset, propensity to involve the proximal colon, and high frequency of multiple CRCs, liver cancer more frequent than endometrial cancer. Chinese HNPCC showed relatively frequent germline mutation of mismatch repair (MMR) genes that correlated closely with high-level MSI and loss of expression of MMR genes protein. PMID- 15786549 TI - One-year follow-up study of Helicobacter pylori eradication rate with 13C-urea breath test after 3-d and 7-d rabeprazole-based triple therapy. AB - AIM: To investigate the long-term role of a 3-d rabeprazole-based triple therapy in patients with Helicobacter pylori(H pylori)-infected active peptic ulcers. METHODS: We prospectively studied 115 consecutive patients with H pylori-infected active peptic ulcers. H pylori infection was confirmed if any two of H pylori DNA, histology, and rapid urease test were positive. Patients were assigned to either an open-labeled 3-d course of oral amoxicillin 1 000 mg b.i.d., clarithromycin 500 mg b.i.d., and rabeprazole 20 mg b.i.d., or 7-d course of oral amoxicillin 1 000 mg b.i.d., clarithromycin 500 mg b.i.d., and rabeprazole 20 mg b.i.d. Subsequently, all patients received oral rabeprazole 20 mg once daily until the 8th wk. Three months after therapy, all patients were followed-up endoscopically for the peptic ulcer, H pylori DNA, histology, and rapid urease test. One year after therapy, H pylori infection was tested using the 13C-urea breath test. RESULTS: The ulcer healing rates 3 mo after therapy were 81.0% vs 75.4% for the 3-d and 7-d groups [intention-to-treat (ITT) analysis, P = 0.47] respectively, and 90.4% vs 89.6% for the 3-d and 7-d groups [per-protocol (PP) analysis, P = 0.89] respectively. The eradication rates 3 mo after therapy were 75.9% vs 73.7% for the 3-d and 7-d groups (ITT, P = 0.79) respectively, and 84.6% vs 87.5% for the 3-d and 7-d groups (PP, P = 0.68) respectively. One year after therapy, seventy-five patients returned to receive the 13C-urea breath test, and the eradication rates were 78.4% vs 81.6% in 3-d and 7-d groups (PP, P = 0.73) respectively. CONCLUSION: Our study showed the eradication rates against H pylori infection 3 and 12 mo after triple therapy were not different between the 3-d and 7-d rabeprazole-based groups. Therefore, the 3-d rabeprazole-based triple therapy may be an alternative treatment for peptic ulcers with H pylori infection. PMID- 15786550 TI - Gallbladder motor function, plasma cholecystokinin and cholecystokinin receptor of gallbladder in cholesterol stone patients. AB - AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease. METHODS: Gallbladder motility was studied by ultrasonography in 33 patients with gallbladder stone and 10 health subjects as controls. Plasma CCK concentration was measured by radioimmunoassay in fasting status (CCK-f) and in 30 min after lipid test meal (CCK-30). Radioligand method was employed to analyze the amount and activity of CCK-R from 33 gallstone patients having cholecystectomy and 8 persons without gallstone died of severe trauma as controls. RESULTS: The percentage of cholesterol in the gallstone composition was more than 70%. The cholesterol stone type was indicated for the patients with gallbladder stone in this study. Based on the criterion of gallbladder residual fraction of the control group, 33 gallstone patients were divided into two subgroups, contractor group (14 cases) and non-contractor group (19 cases). The concentration of CCK-30 was significantly higher in non contractor group than that in both contractor group and control group (55.86+/ 3.86 pmol/L vs 37.85+/-0.88 pmol/L and 37.95+/-0.74 pmol/L, P<0.01), but there was no difference between contractor group and control group. Meanwhile no significant difference of the concentration of CCK-f could be observed among three groups. The amount of CCK-R was lower in non-contractor group than those in both control group and contractor group (10.27+/-0.94 fmol/mg vs 24.59+/-2.39 fmol/mg and 22.66+/-0.55 fmol/mg, P<0.01). The activity of CCK-R shown as KD in non-contractor group decreased compared to that in control group and contractor group. Only was the activity of CCK-R lower in contractor group than that in control group. The ejection fraction correlated closely with the amount of CCK-R (r = 0.9683, P<0.01), and the concentration of CCK-30 correlated negatively with the amount of CCK-R closely (r = -0.9627, P<0.01). CONCLUSION: The distinctive interactive relationship of gallbladder emptying, plasma CCK and CCK-R in gallbladder from this study suggested that the defect of CCK-R may be a key point leading to the impairment of gallbladder motor function and the pathogenesis of cholesterol gallstone formation may differ in two subgroups of gallstone patient, gallbladder non-contractor group or contractor group. PMID- 15786551 TI - HBVPathDB: a database of HBV infection-related molecular interaction network. AB - AIM: To describe molecules or genes interaction between hepatitis B viruses (HBV) and host, for understanding how virus' and host's genes and molecules are networked to form a biological system and for perceiving mechanism of HBV infection. METHODS: The knowledge of HBV infection-related reactions was organized into various kinds of pathways with carefully drawn graphs in HBVPathDB. Pathway information is stored with relational database management system (DBMS), which is currently the most efficient way to manage large amounts of data and query is implemented with powerful Structured Query Language (SQL). The search engine is written using Personal Home Page (PHP) with SQL embedded and web retrieval interface is developed for searching with Hypertext Markup Language (HTML). RESULTS: We present the first version of HBVPathDB, which is a HBV infection-related molecular interaction network database composed of 306 pathways with 1 050 molecules involved. With carefully drawn graphs, pathway information stored in HBVPathDB can be browsed in an intuitive way. We develop an easy-to-use interface for flexible accesses to the details of database. Convenient software is implemented to query and browse the pathway information of HBVPathDB. Four search page layout options-category search, gene search, description search, unitized search-are supported by the search engine of the database. The database is freely available at http://www.bio-inf.net/HBVPathDB/HBV/. CONCLUSION: The conventional perspective HBVPathDB have already contained a considerable amount of pathway information with HBV infection related, which is suitable for in-depth analysis of molecular interaction network of virus and host. HBVPathDB integrates pathway data-sets with convenient software for query, browsing, visualization, that provides users more opportunity to identify regulatory key molecules as potential drug targets and to explore the possible mechanism of HBV infection based on gene expression datasets. PMID- 15786553 TI - Post-radiation survival time in hepatocellular carcinoma based on predictors for CT-determined, transarterial embolization and various other parameters. AB - AIM: In this retrospective study of unresectable hepatocellular carcinoma (HCC), we have investigated the efficacy of CT-derived parameters, laboratory measurements, clinical assessment and associated transarterial embolization (TAE) as predictors of post-radiotherapy survival time. METHODS: Sixty-six patients diagnosed with unresectable HCC that had undergone radiotherapy at two medical university hospitals in Taipei were enrolled in the study. Using multivariant analysis, pre-treatment parameters including tumor number and CT confirmation of PVT and ascites were compared. Multivariant analysis was also used for comparison of the mean pretreatment values for laboratory measurements, including alpha fetoprotein, direct/total bilirubin and GOT/GPT levels, and clinical history of chronic hepatitis across the three survival-time categories. The chi2 was used to test the significance of the relationship between survival time and TAE procedure. The P values for the above tests were deemed statistically significant where P<0.05. RESULTS: Portal vein thrombosis (P = 0.032) and ascites (P<0.05) were negative predictors of post-radiation survival time. Low-grade liver cirrhosis (A or B), lower tumor volume and low levels of AFT, GOT/GPT, and total bilirubin were predictors of longer post-radiation survival time (P<0.05). CONCLUSION: The CT and clinical and laboratory assessment provide a reference for, and enable estimation of, probable survival times in HCC patients after radiotherapy. Tumor volume, severity of liver cirrhosis, status with respect to portal vein thrombosis and ascites and AFT, GOT/GPT and total bilirubin values were significant predictors of survival in this study. PMID- 15786552 TI - Effect of indomethacin on cell cycle proteins in colon cancer cell lines. AB - AIM: To study the effect of indomethacin (IN) on human colon cancer cell line SW480 with p53 mutant and SW480 transfected wild-type p53 (wtp53/SW480) in vitro and investigate molecular mechanism of anti-tumor effect of IN on colon cancer. METHODS: SW480 cells and wtp53/SW480 cells were treated with different concentrations of IN respectively, the expressions of CDK2, CDK4 and p21WAF1/CIP1 protein were detected by Western blotting. RESULTS: IN gradually down-regulated the expression of CDK2, CDK4 protein of wtp53/SW480 cells in a dose-dependent manner, and inhibitory effect reached the maximum level at 600 micromol/L; IN up regulated the expression of p21 WAF1/CIP1 protein in a dose-dependent manner at a certain concentration range, and the expression reached the maximum level at 400 micromol/L, and returned to the base level at 600 micromol/L. The expression of CDK2, CDK4 and p21WAF1/CIP1 protein of SW480 cells did not change. CONCLUSION: IN exerts antitumor effect partly through down regulation of the expression of CDK2, CDK4 protein and up regulation of the expression of p21WAF1/PIC1. PMID- 15786554 TI - Correlation of CD95 and soluble CD95 expression with acute rejection status of liver transplantation. AB - AIM: To analyze the expression levels of soluble form of CD95, CD95 ligand (sCD95 and sCD95L, respectively) in plasma and CD95 expression on CD3+ cells in liver transplanted recipients with acute rejection (AR). METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from 30 clinically liver transplanted recipients. CD95 expression on CD3+ cells was quantitatively measured by two color fluorescence activated cell sorter (FACS) analysis. Lymphocyte surface phenotypes of CD4, CD8, CD16 and CD56 were determined by flow cytometry. Plasma levels of sCD95 and sCD95L were detected by Enzyme Linked-Immuno-Sorbent Assay (ELISA). The results were compared with that from normal healthy volunteers (n = 15 individuals). RESULTS: FACS analysis showed that CD95 expression on CD3+ T cells was significantly increased in liver transplanted recipients with AR compared to that in stable recipients without rejection and infection or healthy individuals who did not undergo transplantation (18,676.93+/-11,588.34/molecule, 6,848.20+/-1 712.96/molecule, 6,418.01+/-2,001.95/molecule, respectively, P<0.01). Whereas no significant difference was seen between liver-transplanted stable recipients and healthy individuals. Furthermore, no significant differences were detected between each group with CD4/CD8 ratio or the percentage of CD16+56+ cells. Plasma levels of sCD95 were significantly higher in transplanted recipients with AR compared to that in stable recipients or healthy individuals (391.88+/-196.00, 201.37+/-30.30, 148.83+/-58.25 pg/mL, respectively, P<0.01). In contrast, the plasma levels of sCD95L in liver- transplanted recipients were not significantly different from that in healthy individuals. CONCLUSION: The present results indicate that the increased CD95 expression on CD3+ cells and the increased levels of sCD95 in plasma may modify the immunological situation of the recipients after transplantation or represent the ongoing graft rejection. PMID- 15786555 TI - Expression of hypoxia-inducible factor 1alpha and vascular endothelial growth factor in hepatocellular carcinoma: Impact on neovascularization and survival. AB - AIM: To study the expression of hypoxia-inducible factor 1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) in hepatocellular carcinoma (HCC) and the impact on neovascularization and survival. METHODS: Expressions of HIF 1alpha, VEGF and microvessel density (MVD) are studied through immunohistochemistry in 36 cases of HCC and the corresponding paraneoplastic tissue and 6 cases of normal liver tissue. The relationship of the expressions of HIF-1alpha and VEGF with the clinicopathological data and survival are analyzed. RESULTS: The positive rate of VEGF in HCC was 32/36, which is significantly higher than that in paraneoplastic tissue and normal liver tissue (P<0.05). The expression of HIF-1alpha in HCC tissue is 24/36, also higher than that in paraneoplastic tissue and normal liver tissue (P<0.05). The expression of VEGF and HIF-1alpha in HCC with microscopic venous invasion is significantly higher than that in HCC without microscopic venous invasion (P<0.05). Spearman correlation analysis does not only show the expression of HIF-1alpha as correlated with the expression of VEGF (r(s) = 0.459, P<0.01), but it also shows the expression of HIF-1alpha and VEGF as correlated with MVD (r(s) = 0.412 and 0.336, respectively, P<0.05). The differences of the survival rates among VEGF positive group and VEGF negative group are significant (P<0.05), whereas the differences of the survival rates among the HIF-1alpha negative group and positive group are not significant (P>0.05). CONCLUSION: HIF-1alpha plays important roles in neovascularization in HCC possibly through regulation of VEGF transcription. PMID- 15786556 TI - Application of atomic force microscopy in blood research. AB - AIM: To find suitable solutions having lesser granules and keeping erythrocytes in normal shapes under atomic force microscopy (AFM). METHODS: Eight kinds of solutions, 1% formaldehyde, PBS buffer (pH7.2), citrate buffer (pH 6.0), 0.9% NaCl, 5% dextrose, TAE, 1640 medium and 5% EDTA-K2, were selected from commonly used laboratory solutions, and venous blood from a healthy human volunteer was drawn and anticoagulated with EDTA-K2. Before scanned by AFM (NanoScopeIIIa SPM, Digital Instruments, Santa Barbara, CA), a kind of intermixture was deposited on freshly cleaved mica and then dried in the constant temperature cabinet (37 degrees). RESULTS: One percent formaldehyde, citrate buffer, 5% dextrose, TAE, were found to keep human erythrocytes in normal shape with few particles. Processed by these solutions, fine structures of human erythrocyte membrane were obtained. CONCLUSION: One percent formaldehyde, citrate buffer, 5% dextrose and TAE may be applied to dispose erythrocytes in AFM. The results may offer meaningful data for clinical diagnosis of blood by AFM. PMID- 15786557 TI - 14C-urea breath test in patients undergoing anti-tuberculosis therapy. AB - AIM: Urea breath test (UBT) is a non-invasive diagnostic test for detecting the presence of Helicobacter pylori (H pylori). In this study we evaluated the effect of anti-tuberculosis therapy on the results of 14C-UBT. METHODS: Patients, with the diagnosis of tuberculosis (TB) who had a positive UBT at the point of starting anti-TB therapy, were included. None had a history of peptic ulcer disease or had taken antibiotics, bismuth compounds and/or PPI in the previous month. 14C-UBT was repeated at the end of the second month and the end of treatment period and one month after completion of treatment course. RESULTS: Thirty-five patients (23 males) were enrolled. 14C-UBT was negative in all 35 patients (100%) at the end of the second month and remained negative in 30 cases (85.7%) at the end of the treatment course. One month after completion of treatment course, UBT remained negative in 13 patients (37.1%). CONCLUSION: Our report underscores the need for caution while interpreting urea breath test results in patients undergoing anti-TB therapy. Furthermore, the combination of drugs used in this study resulted in H pylori eradication in a minority of patients. PMID- 15786558 TI - Sacral anterior root stimulated defecation in spinal cord injuries: an experimental study in canine model. AB - AIM: To investigate whether there was a dominant sacral root for the motive function of rectum and anal sphincter, and to provide an experimental basis for sacral root electrically stimulated defecation in spinal cord injuries. METHODS: Eleven spinal cord injured mongrel dogs were included in the study. After L4-L7 laminectomy, the bilateral L7-S3 roots were electrostimulated separately and rectal and sphincter pressure were recorded synchronously. Four animals were implanted electrodes on bilateral S2 roots. RESULTS: For rectal motorial innervation, S2 was the most dominant (mean 15.2 kPa, 37.7% of total pressure), S1 (11.3 kPa, 27.6%) and S3 (10.9 kPa, 26.7%) contributed to a smaller part. For external anal sphincter, S3 (mean 17.2 kPa, 33.7%) was the most dominant, S2 (16.2 kPa, 31.6%) and S1 (14.3 kPa, 27.9%) contributed to a lesser but still a significant part. Above 85% L7 roots provided some functional contribution to rectum and anal sphincter. For both rectum and sphincter, the right sacral roots provided more contribution than the left roots. Postoperatively, the 4 dogs had electrically stimulated defecation and micturition under the control of the neuroprosthetic device. CONCLUSION: S2 root is the most dominant contributor to rectal pressure in dogs. Stimulation of bilateral S2 with implanted electrodes contributes to good micturition and defecation in dogs. PMID- 15786559 TI - Endoscopic ultrasonographic appearance of gastric emphysema. AB - Emphysematous gastritis (or phlegmonous gastritis) and gastric emphysema (or gastric pneumatosis) are variations of conditions associated with the presence of intramural air in the stomach. The presence of air in the gastric wall is a very rare clinical condition, associated with bacterial infection, increased intragastric pressure from gastric outlet obstruction, gastric mucosal disruption or air dissection from the mediastinum. In adults, this can occur in the setting of instrumentation-related injury, gastric outlet obstruction by gastric, duodenal or pancreatic malignancies or bowel ischemia. Here we describe a case of gastric emphysema related to repeated biliary stenting and partial duodenal obstruction in a patient with inoperable periampullary cancer, and provide the first description of the endoscopic ultrasonographic findings of gastric emphysema in the literature. In our case, endoscopic ultrasound showed a band of bright echogenicity arising from the submucosa layer, representing air in the gastric wall. PMID- 15786560 TI - Actinomycosis mimicking recurrent carcinoma after Whipple's operation. AB - Actinomycosis is a rare, chronic, spreading, suppurative, granulomatous and fibrosing infection. Actinomyces are normal inhabitants of the oral cavity and gastrointestinal tract. They rarely cause disease and are seldom reported as pathogens. Herein, we reported on a 69-year-old male patient who had undergone Whipple's operation due to ampulla Vater carcinoma, and became infected with actinomycosis at the pancreaticojejunostomy, which mimicked a recurrent malignancy. He was treated with radical resection of the mass at the pancreaticojejunostomy and had an uneventful postoperative course. PMID- 15786562 TI - Fatal thrombotic complications of hepatic cystic compression of the inferior vena: a case report. AB - Of 5% of patients who develop liver cysts, only 10-15% of them come for medical attention, typically because of dull right upper quadrant pain, abdominal bloating or early satiety. We treated a 77-year-old female with a rare complication of inferior vena cava thrombosis. The patient expired due to septic shock and multiple organ failure. PMID- 15786561 TI - Candidal liver abscesses and cholecystitis in a 37-year-old patient without underlying malignancy. AB - We report a case of candidal liver abscesses and concomitant candidal cholecystitis in a diabetic patient, in whom differences were noted relative to those found in patients with hematologic malignancies. In our case, the proposed entry route of infection is ascending retrograde from the biliary tract. Bile and aspirated pus culture repeatedly tested positive, and blood negative, for Candida albicans and Candida glabrata. Cholecystitis was cured by percutaneous gallbladder drainage and amphotericin B therapy. The liver abscesses were successfully treated by a cumulative dosage of 750 mg amphotericin B. We conclude that in cases involving less immunocompromised patients and those without candidemia, a lower dosage of amphotericin B may be adequate in treating candidal liver abscesses. PMID- 15786563 TI - What is the role of mitomycin C in advanced gastric cancer? PMID- 15786564 TI - Bibliography. Current world literature. Rehabilitation medicine in rheumatic diseases. PMID- 15786565 TI - Inflammatory pseudotumor of the sigmoid colon mesentery: US and CT findings (2004:12b). AB - The concept of inflammatory pseudotumor has evolved from meticulous pathological studies; some of its histological features resemble a spindle-cell sarcoma. Despite the fact that it usually affects children and young adults, only limited numbers of childhood cases have been reported in the pediatric literature. Recognition of this rare entity is important because the clinical manifestations and radiological features may be indistinguishable from a malignant lymphoproliferative disorder. This entity has been reported to be anywhere in the body, including a variety of intra-abdominal organs. Although one of the most common intra-abdominal sites is the mesentery, localization within the mesentery of the sigmoid colon is particularly rare. We present a case of abdominal inflammatory pseudotumor of the sigmoid colon mesentery, defining its radiological and primarily ultrasound and Doppler ultrasound findings, with a review of additional examples from the literature. PMID- 15786567 TI - Sonja Buckley. PMID- 15786568 TI - Shock-wave therapy for plantar fasciitis. PMID- 15786570 TI - PA: Agency home aide killed driving home: workers compensation death benefits issue. Le v. Workers' Compensation Appeal Board, No 1815 C.D. 2004 (Pa. Commw. 01/27/2005). PMID- 15786569 TI - The mechanistic classification of CCVM. PMID- 15786571 TI - Use of bone marrow examination in HIV/AIDS. PMID- 15786572 TI - HIV-hepatitis C virus coinfection: questions and answers. PMID- 15786573 TI - Acute presentation of mania. PMID- 15786574 TI - Author's addendum to November 2004 clinical challenge. PMID- 15786575 TI - Male circumcision and the risk of HIV infection. AB - Epidemiologic data have suggested that male circumcision is a major protective factor against male heterosexual HIV transmission and may explain the significant geographic differences in the prevalence of HIV observed within sub-Saharan Africa. To assess the evidence of the protective effect of male circumcision, African studies on its association with HIV infection were reviewed. These studies' systematic lack of control of important confounding factors makes the assessment of the association between male circumcision and HIV transmission very difficult and raises doubt about the validity of the current findings. Randomized trials are needed to determine the true strength of the association. Until then, a decision to recommend mass male circumcision to prevent HIV transmission in sub Saharan Africa is premature and risky. PMID- 15786576 TI - Editorial comment: male circumcision to reduce HIV acquisition--not quite yet. PMID- 15786577 TI - Neurologic manifestations of HIV infection: an Indian hospital-based study. AB - Neurologic manifestations affecting the nervous system at all levels and stages of HIV infection are common and increasing with the extended survival of HIV positive persons. A study was conducted to document neurologic events in HIV cases in India, a country in which persons are primarily infected with subtype C HIV-1. The study's aims were to evaluate the incidence of admissions due to HIV related neurologic events, to study the diverse clinical presentations of neurologic abnormalities, and to correlate these with the CD4 count. The demographic and clinical data of 1606 HIV-positive patients were analyzed by a retrospective review of the case records. In 411 patients, 486 single or coexisting neurologic manifestations were found--an overall prevalence of 25.6%, which ranged from 15.8% in 1993 to 26.6% in 2003. Meningitis was the most common CNS infection in patients with HIV/AIDS (39.4%), followed by mass lesions (27.5%). Most of these complications were observed in the advanced stages of infection. CD4 counts were performed in 29.9% of the cases with neurologic events, mainly on those seen since 2000, and the mean CD4 count was 89 cells/microL. This study revealed not only the high prevalence of various neurologic events but also their nature, clinical presentation, and symptoms. The results underscore the need for precise diagnosis and parallel assessment of CD4 counts, which is important for the practical management of an specific therapy for the HIV-infected patient. PMID- 15786578 TI - Editorial comment: deja vu--neurologic complications of HIV infection in the expanding epidemic. PMID- 15786579 TI - Use of porcine renal capsule matrix as a full-thickness dermal wound-healing material in rats. AB - OBJECTIVE: To compare the utility of porcine renal capsule matrix (RCM) with porcine small intestinal mucosa (SIS) in a rat full-thickness skin wound model. METHOD: Groups of rats had surgically-created wounds filled with either SIS or RCM. On each rat a contralateral wound was left unfilled (RCM-U or SIS-U). Wound diameter was measured 3, 7, 12, 17, 26 and 30 days after creation. Wound sites sampled 3, 7, 14, 28, 42 and 56 days after wound creation were numerically graded for degree of histologic change and for collagen content, based on intensity of trichrome staining. RESULTS: Wounds in all groups rapidly contracted to less than 50% of the original diameter within 12 days. There were no differences in wound diameter between RCM- and SIS-treated wounds at any time point, but these wounds had significantly greater (p < 0.001) diameters than their unfilled counterparts on days 7, 12 and 17. There were no differences in histologic scores or trichrome staining scores between RCM- and SIS-treated wounds and their unfilled counterparts at any time point, except for a greater (p < 0.05) histologic score in SIS-treated wounds compared with unfilled controls on day 14. In both treatment groups an acute inflammatory response at the wound site was soon replaced by an influx of macrophages and fibroblasts. CONCLUSION: The results show that RCM is equivalent to SIS for the treatment of full-thickness wounds and that these materials may enhance wound healing in terms of wound-tissue collagenisation and maturation. These materials therefore merit further study in other wound-care models. PMID- 15786581 TI - Interim UK guidelines for management of close community contacts of invasive group A streptococcal disease. AB - Group A streptococci cause a wide range of illnesses from non-invasive disease such as pharyngitis to more severe invasive infections such as necrotising fasciitis. There remains uncertainty about the risk of invasive disease among close contacts of an index case of invasive disease and whether this risk warrants antibiotic prophylaxis. A 19-200 fold increased risk among household contacts has been reported in the literature. Recommendations for antibiotic prophylaxis regimens vary by country. A comprehensive literature review together with preliminary analysis of 2003 United Kingdom data from the strep-EURO programme informed the interim recommendations of an expert working group. The evidence base to formulate definitive guidance is weak. Risk calculations based on provisional UK data estimated that over 2,000 contacts would need to receive antibiotic prophylaxis to prevent a subsequent case of invasive group A streptococcal disease. The Working Group considered that currently available evidence did not warrant the routine administration of chemoprophylaxis to all close community contacts. More robust risk estimates will be derived from ongoing UK surveillance data to inform a review of this guidance in 2005. PMID- 15786582 TI - Preventing person-to-person spread following gastrointestinal infections: guidelines for public health physicians and environmental health officers. AB - This guidance updates advice on preventing person-to-person spread of gastrointestinal infections in the general population, first published in 1983 by the Public Health Laboratory Service and last updated in 1995. It represents a consensus of informed opinion and is particularly aimed at those public health physicians and environmental health officers who do not specialise in communicable disease control. It addresses predominantly the organisms that more commonly present them with problems. The guidance considers general measures, enteric precautions, exclusion from work, school and other settings and groups that pose an increased risk of spreading infection. PMID- 15786580 TI - Effect of byproducts from the ozonation of pyrene: biphenyl-2,2',6,6' tetracarbaldehyde and biphenyl-2,2',6,6'-tetracarboxylic acid on gap junction intercellular communication and neutrophil function. AB - In this study, biphenyl-2,2',6,6'-tetracarbaldehyde, an initial byproduct formed from the ozonation of pyrene, and biphenyl-2,2',6,6'-tetracarboxylic acid, a subsequent pyrene ozonation byproduct, were evaluated using two toxicology assays to compare the toxicity of ozonation byproducts with that of the parent compound. The first assay measured the potential for the compounds to block gap junctional intercellular communication (GJIC) using the scrape loading/dye transfer technique in normal WB-344 rat liver epithelial cells. The second assay evaluated the ability of the compounds to affect neutrophil function by measuring the production of superoxide in a human cell line (HL-60). Pyrene significantly blocked intercellular communication (f = 0.2-0.5) at 40 microM and complete inhibition of communication (f < 0.2) occurred at 50 microM. Gap junctional intercellular communication in cells exposed to biphenyl-2,2',6,6' tetracarbaldehyde reached f < 0.5 at a concentration of 15 microM. At concentrations greater than 20 microM, biphenyl-2,2',6,6'-tetracarbaldehyde was cytotoxic and the inhibition of GJIC was caused by cell death. Biphenyl-2,2',6,6' tetracarboxylic acid was neither cytotoxic nor inhibitory to GJIC at the concentrations tested (10-500 microM). Exposure to biphenyl-2,2',6,6' tetracarbaldehyde resulted in a concentration-dependent decrease in phorbol 12 myristate 13-acetate-stimulated O2- production. Neither exposure to pyrene nor biphenyl-2,2',6,6'-tetracarboxylic acid caused a significant toxic effect on neutrophil function. PMID- 15786583 TI - [Antidiabetic activity of the leaves of Vemonia colorata (Wilid.) Drake (Composees) in alloxan-induced diabetic rats]. AB - Vemonia colorata is distributed through Africa (Benin, Cameroon, Senegal, Togo etc...). Its leaves are commonly used by african tradipractitioners for treating diabetes. However, the antidiabetic activity of the leaves of V. colorata had never been investigated in experimental conditions. The present study aimed to test the aqueous extract of the leaves of V. colorata for its effects in normoglycaemic and alloxan-induced diabetic rats in comparison to glibenclamide antidiabetic activity. Such glibenclamide, the aqueous extract of V. colorata (300 mg/kg, per os) induced a significant hypoglycaemic effect in normoglycaemic rats. The blood glucose varied from 0.77 +/- 0.01 to 0.58 +/- 0.01 g/l (p < 0.05, n = 5). It also reduced significantly the fasting glucose level of the hyperglycaemic rats induced with oral administration of glucose (4 g/kg). In alloxan-induced diabetic rats, glibenclamide (0.2 mg/kg, per os) lowered significantly the blood glucose from 2.40 +/- 0.30 to 0.70 +/- 0.40 g/l (p < 0.05, n = 5). As well as glibenclamide, the aqueous extract of V. colorata (300 mg/kg, per os) decreased the blood glucose in alloxanic rats from 2.80 +/- 0.10 to 1.00 +/- 0.20 g/l (p < 0.05, n = 5). The aqueous extract of the leaves of V. colorata possesses both hypoglycaemic and antidiabetic effect in normoglycaemic and alloxan-induced diabetic rats. This may indicate the ethnopharmacological basis of the use of V. colorata leaves in traditional medicine for treating diabetes. PMID- 15786584 TI - For the dental patient. Dental implants. An option for replacing missing teeth. PMID- 15786585 TI - Patient notes. Premenstrual syndrome. PMID- 15786586 TI - Relations in color among maxillary incisors and canines. AB - OBJECTIVES: To establish the strength of color relation among the maxillary central incisor, lateral incisor and canine teeth by using digital photography. This relation might be useful for the color reconstruction of the missing part of a tooth from the color obtained from neighboring teeth. METHODS: The L*a*b* values of the maxillary incisors and canines were determined in 100 subjects. Paired t-tests, Pearson correlations and linear regression analyses were used to describe the relation of the L*a*b* values between the three teeth for each segment (cervical, middle and incisal). RESULTS: The mean L*a*b* values of the canines differed statistically significantly from the values of the central incisors (p<0.001) except for L* and a* values in the cervical segment between the lateral incisor and canine. The color differences between the incisors were statistically significant in fewer cases. The relation in color was the highest between the cervical segments of the three teeth, with respect to L* (r = 0.45 0.65; p < 0.001), b* (r = 0.49-0.55; p < 0.001), and a* (r = 0.38-0.61; p < 0.001), and no relation was found between the incisal segments of central incisor and canine. SIGNIFICANCE: There is a relation in color between the maxillary incisors and canines, which is stronger between the cervical than between the middle and incisal segments. Therefore, the color prediction is most reliable when the cervical parts of the teeth are used. PMID- 15786587 TI - Modification of ammonium lignosulfonate by phenolation for use in phenolic resins. AB - The structural modification of softwood ammonium lignosulfonate by phenolation was studied. A central factorial design was applied to determine the influence of reaction conditions (temperature, time and lignosulfonate content) on the properties of phenolation products. The reaction was monitored through the concentration of free phenol determined by gas chromatography (GC). The characterization of phenolation products was accomplished by gel permeation chromatography (GPC), fourier transform infrared (FTIR) and 1H nuclear magnetic resonance (1H NMR). Empirical models were developed to predict phenol conversion and adduct formation according to operating conditions. The optimum phenolation conditions, attained by means of a response surface method, were found to be 120 degrees C; 160 min and 30% lignosulfonate content PMID- 15786588 TI - Medicare program; Medicare prescription drug benefit; interpretation. Final rule; interpretation. AB - This final rule modifies or clarifies our interpretations in several areas of the final rule titled "Medicare Prescription Drug Benefit" published in the Federal Register on January 28, 2005. First, it clarifies our interpretation of "entity", to respond to inquiries we received subsequent to the publication of the Prescription Drug Benefit (Part D) final rule on January 28, 2005. We were asked whether a joint enterprise could be considered an "entity" under section 1860D 12(a)(1) of the Social Security Act (the Act), for purposes of offering a prescription drug plan (PDP). Our interpretation is discussed in the Supplementary Information section of this final rule. Second, also subsequent to the publication of the Prescription Drug Benefit (Part D) final rule on January 28, 2005, we received inquiries from parties about our discussion of the actuarial equivalence standard and the manner in which an employee health plan sponsor could apply the aggregate net value test in the regulatory text of the final rule. Our interpretation is discussed in the "Provisions" section of this final rule. In addition, subsequent to publishing the August 3, 2004 proposed rule (69 FR 46684), we received comments on how the late enrollment penalty would be coordinated with the late enrollment penalty for Part B, and whether the one percent penalty would be sufficient to control for adverse selection. We clarify in the Provisions section of this final rule that the example given in the proposed rule, published on August 3, 2004, did not accord with the proposed or final regulatory language because it did not account for the fact that the base beneficiary premium increases on an annual basis. To remedy this error and in response to comments received on the proposed rule, we provide an interpretation that as the base beneficiary premium increases, the late enrollment penalty must also increase, and is in keeping with how the Part B penalty is calculated. Finally, we are providing clarifying language related to transitioning Part D enrollees from their prior drug coverage to their new Part D plan coverage. The Medicare Prescription Drug Benefit final rule will take effect on March 22, 2005. Our interpretations are deemed to be included in that final rule. PMID- 15786589 TI - Medicare program; establishment of the Medicare Advantage Program; interpretation. Final rule; interpretation. AB - This final rule clarifies our interpretation of the meaning of "entity" in the final rule titled "Medicare Program; Establishment of the Medicare Advantage Program" published in the Federal Register on January 28, 2005 (70 FR 4588). Subsequent to the publication of the Medicare Advantage (MA) final rule on January 28, 2005, we have received inquiries from parties interested in offering an MA Regional Plan concerning whether they could jointly enter into a contract with us to offer a single MA Regional Plan in a multistate region. The participating health plans wish to contract with each other to create a single "joint enterprise." They have asked us whether such a joint enterprise could be considered an "entity" under sections 1859(a)(1)and 1855(a)(1) of the Social Security Act, for purposes of offering an MA Regional Plan. The MA final rule is scheduled to take effect on March 22, 2005. Our interpretation of the word "entity" that follows in the "Supplementary Information" section of this final rule is deemed to be included in that final rule. PMID- 15786590 TI - Medical devices; immunology and microbiology devices; classification of the automated fluorescence in situ hybridization enumeration systems. Final rule. AB - The Food and Drug Administration (FDA) is classifying automated fluorescence in situ hybridization (FISH) enumeration systems into class II (special controls). The special control that will apply to the device is the guidance document entitled "Class II Special Controls Guidance Document: Automated Fluorescence in situ Hybridization (FISH) Enumeration Systems." The agency is classifying the device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. Elsewhere in this issue of the Federal Register, FDA is publishing a notice of availability of a guidance document that is the special control for this device. PMID- 15786591 TI - Hand2 regulates epithelial formation during myocardial diferentiation. AB - Myocardial differentiation is initiated by the activation of terminal differentiation gene expression within a subset of cells in the anterior lateral plate mesoderm. We have previously shown that shortly after this activation, myocardial cells undergo epithelial maturation [1], suggesting that myocardial differentiation encompasses both molecular and cellular changes. To address the question of how the molecular programs driving myocardial gene expression and the formation of the myocardial epithelium are integrated, we analyzed the role of two essential myocardial terminal-differentiation factors, Hand2 and Gata5, in myocardial epithelia formation. hand2 and gata5 mutants exhibit a much-reduced number of myocardial cells and defects in myocardial gene expression [2,3]. We find that the few myocardial precursors that are present in hand2 mutants do not polarize. In contrast, embryos with reduced Gata5 function exhibit polarized myocardial epithelia despite a similar reduction in myocardial precursor number, indicating that proper cell number is not required for epithelial formation. Taken thogether, these results indicate that Hand2 is uniquely required for myocardial polarization, a previously unappreciated role for this critical transcription factor. Furthermore, these results demonstrate that two independent processes, the polarizaton of myocardial precursors and the allocation of proper cell number, contribute to myocardial development. PMID- 15786592 TI - Ethological analysis of a polar team in the French Antarctic station Dumont d'Urville as simulation of space teams for future interplanetary missions. AB - This new ethological study focuses on the co-adaptation of the crew's spatial behavior to social isolation in a polar base thus simulating long-term living and working of a space team. The method consisted in drawing the subjects' position (n=13) on an observation map at the midday and evening meals at the Dumont d'Urville French station in Antarctica, daily during the summer campaigns and weekly during the winter-over of the TA46 mission. Quantitative data are presented in geocentric (positions), allocentric (distances) and egocentric (orientations) analyses with an emphasis on three adaptative periods (first 3 months, intermediary 2 months and last 3 months of isolation). Results show a large space occupancy during the first week after arrival and the last week before departure from the polar base, and a team-members' grouping during the winter-over. On the over-all time, the inter-individual distances increase. The social orientations are higher at the beginning than at the end of the mission. Discussion underlies the pertinent use of such ethological indicators collected from polar stations as predictors of well-being and optimal-working of the future orbital and planetary stations users. PMID- 15786593 TI - Planning, implementation and optimization of future space missions using an immersive visualization environment (IVE) machine. AB - Beginning in 1995, a team of 3-D engineering visualization experts assembled at the Lockheed Martin Space Systems Company and began to develop innovative virtual prototyping simulation tools for performing ground processing and real-time visualization of design and planning of aerospace missions. At the University of Colorado, a team of 3-D visualization experts also began developing the science of 3-D visualization and immersive visualization at the newly founded British Petroleum (BP) Center for visualization, which began operations in October, 2001. BP acquired ARCO in the year 2000 and awarded the 3-D flexible IVE developed by ARCO (beginning in 1990) to the University of Colorado, CU, the winner in a competition among 6 Universities. CU then hired Dr. G. Dorn, the leader of the ARCO team as Center Director, and the other experts to apply 3-D immersive visualization to aerospace and to other University Research fields, while continuing research on surface interpretation of seismic data and 3-D volumes. This paper recounts further progress and outlines plans in Aerospace applications at Lockheed Martin and CU. PMID- 15786594 TI - Biological nanostructures: platforms for analytical chemistry at the sub zeptomolar level. AB - The analysis and manipulation of molecules at the sub-zeptomolar level (i.e., from 1 to 600 molecules) remains the unconquered frontier of analytical chemistry. While some techniques offer sensitivity to single molecules, there are no established tools for the manipulation of such small quantities of material. Scanning probe lithography has begun to provide practicable means to manipulate biological organisation on length scales of 100 nm and less, and three promising approaches (dip-pen nanolithography, nanoshaving, and scanning near-field photolithography) are reviewed. Each offers extraordinary spatial resolution combined with the capability for use under ambient and, in some cases, fluid conditions. These techniques offer a multitude of strategies that may at last make the manipulation of handfuls of molecules--and perhaps single molecules--a practical possibility for the analytical chemist. PMID- 15786595 TI - Occupational history as the key to the recognition and prevention of workplace related lung disease. AB - Occupational and environmental exposures cause substantial lung disease morbidity and mortality. A detailed exposure history is the key to detecting and treating these largely preventable illnesses and deaths. Components of an exposure history include details of the current and past work histories as well as of nonoccupational exposures. Identification of a sentinel health event requires a public health approach to the follow-up of affected individuals. This approach includes industrial hygiene sampling and monitoring, engineering and administrative controls, institution of proper work practices, education, and medical monitoring. PMID- 15786596 TI - The pneumoconioses. AB - Most recent studies have indicated that the incidence and prevalence of the mineral pneumoconioses have been declining as dust control has improved, This finding is particularly true for coal workers' pneumoconiosis, asbestosis, and silicosis. Although the present preoccupation with molecular biology and immunology has led to a spate of studies of the mechanisms involved in the development of these conditions, and although many advances have been made, no convincing complete elucidation of these mechanisms has evolved. Meanwhile, the "paratoxicologic fringe," supported by the National Institute of Occupational Safety and Health and the Environmental Protection Agency, has been presenting the notion that almost every naturally occurring or synthesized mineral ranging from aluminum to silica is a carcinogen. Much of the evidence produced by this consortium has been found wanting, especially in regard to the alleged hazards of talc, beryllium, and silica. PMID- 15786597 TI - Air pollution. AB - Despite progress in improving ambient air quality and reducing personal exposures to air pollutants in indoor settings, population exposures to air pollutants remain widespread and are an important public health concern. Many air pollutants are associated with adverse health effects; the most serious include development and exacerbation of asthma, loss of lung function, impaired neurobehavioral development, and death. These outcomes are the focus of current health effects research on environmental tobacco smoke and radon in indoor settings, and lead, ozone, and particulate matter outdoors. Advances in methods to measure exposures to specific pollutants and the identification of markers of individual susceptibility are providing improved data on the relation of exposure and disease, and, in turn, will improve efforts to protect public health. PMID- 15786598 TI - Burns and smoke inhalation. AB - Smoke inhalation injury affects nearly one third of all major burn victims. Significant inhalation exposures must be suspected in persons who were entrapped in a closed space or who became unconscious during a fire. Each individual fire generates a characteristic smoke depending on the type of materials burnt, temperatures reached during pyrolysis, and the availability of oxygen to sustain combustion. In addition to variable amounts of thermal loads, firesmoke may contain mixtures of carbon monoxide, hydrogen cyanide, nitrogen oxides, and other highly irritating gases. Each constituent of firesmoke may potentially create pulmonary and systemic toxicities and must be considered in every victim of smoke inhalation. PMID- 15786599 TI - Toxic gas inhalation. AB - Severe toxic fume inhalations, usually accidental due to human error or equipment failure, can result in immediate death from asphyxia or cause mild to severe respiratory distress from acute upper airways inflammation, delayed pulmonary edema, respiratory muscle dysfunction, or a combination of illnesses. Most patients are expected to survive and recover with little or no residual dysfunction regardless of the severity of the initial event. However, in some cases disabling long-term sequelae, eg, bronchiectasis, chronic airflow obstruction, bronchial hyperreactivity, asthma-like disease (reactive airways dysfunction syndrome), bronchiolitis obliterans, or residual psychophysiologic dyspnea, can occur. Therapy of the respiratory effects of irritant gases should follow the general principles used for the treatment of upper and lower airway obstruction, noncardiogenic pulmonary edema, and hemorrhagic pneumonitis while spontaneous healing and recovery occurs, because no specific therapy is available for direct chemical pulmonary injury. Corticosteroids are frequently used and recommended, but their efficacy in altering the course and outcome of respiratory injury has not yet been properly documented. PMID- 15786600 TI - Bronchiolitis with airflow obstruction. AB - Bronchiolar lesions are becoming increasingly recognized as an important cause of airflow obstruction. For this reason, it is helpful to have an update of the current clinical, radiographic, and immunologic perspective. Among the bronchiolar airflow disorders, diffuse panbronchiolitis is related to HLA antigen Bw54, and low-dose, long-term erythromycin appears to be effective therapy. Bronchiolitis obliterans can be classified histologically as constrictive bronchiolitis and as proliferative bronchiolitis obliterans. Idiopathic, postfume, postinfectious, and connective tissue disease bronchiolitis obliterans continue to be rare and often have a poor prognosis. Bronchiolitis obliterans associated with lung transplantation is undergoing intensive investigation with regard to pathogenesis, immunologic study, early detection, and treatment. The lesion appears to be a form of chronic organ rejection. The recognition of the distinctive differences among the bronchiolar airflow disorders by clinicians and clinical investigators is essential for improved patient care, for a greater understanding of the pathogenesis of the disorder, and for development of new therapeutic advances. PMID- 15786601 TI - An update on bronchiectasis. AB - Bronchiectasis is associated with heterogeneous predisposing conditions that cause abnormal dilatation and persistent inflammation in the bronchial tree and lung parenchyma. The disease remains a common cause of significant morbidity and mortality, especially when associated with hereditary disorders such as cystic fibrosis, ciliary dyskinesia, and immunodeficiency states. Recent investigations have focused on the inflammatory mediators involved in the pathogenesis of bronchiectasis. High-resolution computed tomography is now the diagnostic modality of choice and may also contribute to clinical management. Computed tomography and high-resolution computed tomography have identified bronchiectasis in individuals with HIV and alpha-1-antitrypsin deficiency. Early identification of predisposing disorders and aggressive management of symptoms has already been demonstrated by prolonged survival and decreased morbidity in cystic fibrosis patients, and similar management may benefit other populations with bronchiectasis. New treatments such as recombinant human DNase have been shown to improve pulmonary function and quality of life in cystic fibrosis patients and may prove useful in other chronic inflammatory lung disease. PMID- 15786602 TI - Pathology of chronic obstructive pulmonary disease. AB - This gathering of new observations about chronic obstructive pulmonary disease, collected under the loosely defined heading of "pathology," creates a certain air of excitement. Vascular engorgement in concert with muscle contraction produces small airways narrowing in asthma, but not in chronic obstructive pulmonary disease. Stenotic small airways can be visualized in three dimensions. Fibrosis may be an integral part of emphysema, stretching its definition somewhat. Microvascular injury seems to produce emphysema. The protease-antiprotease theory of emphysema has competition from the inflammation-repair-fibrosis sequence seen in other organs. The mystery of why some smoker's lungs remain unaffected by tobacco smoke is further documented but unsolved; neuroendocrine cells and their neuropeptides may be important. PMID- 15786603 TI - Management of chronic obstructive pulmonary disease. AB - Chronic obstructive lung disease affects over 15 million people in the United States. The management of this disease in patients is complex, involving antibiotics, mucolytics, bronchodilators, corticosteroids, and rehabilitation. Several studies investigating controversial areas of management were published in 1993 and 1994. New developments in pulmonary rehabilitation, oxygen therapy, noninvasive ventilation, corticosteroids, bronchodilators, vasodilators for pulmonary hypertension, lung transplantation, lung surgery for bullae, and alpha 1-antitrypsin replacement therapy provide physicians with exciting new opportunities for managing chronic obstructive pulmonary disease. PMID- 15786604 TI - Diagnosis of chronic obstructive pulmonary disease and differentiation from asthma. AB - Asthma and chronic obstructive pulmonary disease are both common diseases, which together afflict approximately 25 million Americans. Although expiratory airflow obstruction is the common physiologic abnormality, asthma and chronic obstructive pulmonary disease are characterized by unique pathologic findings, clues from clinical histories, and laboratory test results. Despite some overlap in these characteristics, it is usually possible to differentiate these two conditions. This distinction is important for the healthcare provider to communicate a realistic prognosis to the patient and the patient's family, and to institute appropriate therapy. PMID- 15786605 TI - Tobacco smoking and new developments in public health and clinical interventions. AB - The public policy environment regarding tobacco use in the United States has experienced a dramatic change during the past year. Along with calls for regulatory review of cigarettes, important new scientific information has become available regarding the health effects of environmental tobacco smoke and the efficacy of nicotine replacement therapy, which is used to support smoking cessation efforts. Specifically, recent studies have suggested that environmental tobacco smoke exposure increases risk for coronary heart disease in nonsmoking adults in addition to causing lung cancer and other respiratory diseases. Children are exposed to environmental tobacco smoke at home and in public, resulting in increased risk of bronchitis, pneumonia, bronchial hyperresponsivity, and sudden infant death syndrome. In a climate of increasing concern about the direct and indirect effects of tobacco smoke, three independent meta-analyses concluded that nicotine replacement therapy increased smoking cessation efficacy two- to threefold. In addition, research is beginning to identify factors associated with successful and unsuccessful cessation attempts using nicotine replacement therapy, resulting in the possibility of individualized treatments and clinical interventions designed for maximum efficacy. PMID- 15786606 TI - Bibliography of the current world literature. Obstructive , occupational, and environmental diseases. PMID- 15786607 TI - Summary health statistics for U.S. adults: National Health Interview Survey, 1997. AB - This report presents health statistics from the 1997 National Health Interview Survey for the civilian noninstitutionalized adult population, classified by sex, age, race and Hispanic origin, poverty status, region of residence, and where appropriate, education, income, health insurance coverage, marital status, and place of residence. The topics covered are health status and limitations in activity, health care access and utilization, health behaviors and lifestyle, chronic condition prevalence, and knowledge and attitudes toward the Acquired Immunodeficiency Syndrome (AIDS). Source of Data The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized U.S. population. Data are collected during face-to-face interviews. Highlights Among adults 18 years of age or over, 12% had ever been told by a doctor or health professional that they had heart disease, 6% had ever been told they had coronary heart disease, and 19% had been told on two visits or more that they had hypertension. Nearly a quarter of adults were current smokers, 23% were former smokers, and 52% had never smoked. Sixty-one percent did not engage in any leisure-time vigorous physical activity, while approximately 24% engaged in such activity three times or more per week. Based on estimates of body mass index, 4% of adults were underweight, 42% were at a healthy weight, 35% were overweight, and 19% were obese. PMID- 15786608 TI - Summary health statistics for the U.S. population: National Health Interview Survey, 1997. AB - OBJECTIVES: This report presents health statistics from the 1997 National Health Interview Survey for the civilian noninstitutionalized population of the United States, classified by age, gender, race and Hispanic origin, poverty status, income, education, place of residence, region of residence, and, where appropriate, health insurance coverage. The topics covered are health status and limitations of activity, injuries and poisonings, health care access and utilization, and health insurance coverage. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized U.S. population. Data are collected during face-to-face interviews with adults present at the time of interview. Information about children and absent adults is obtained from an adult proxy respondent. HIGHLIGHTS: Nearly 40% of Americans reported having excellent health in 1997, while almost 10% reported having either fair or poor health. Regarding health insurance coverage, 16% of the U.S. population did not have any health insurance coverage in 1997. Nineteen percent of non-Hispanic black persons and 33% of Hispanics were uninsured in 1997 as opposed to 12% of non-Hispanic white persons. Further, 45% of poor Hispanics and 43% of near poor Hispanics under age 65 years were uninsured, while among persons ages 65 years and over, 7% of poor Hispanics were uninsured. Lastly, 78% of non-Hispanic white persons under age 65 years had private health insurance coverage as opposed to 55% non-Hispanic black persons and 46% of Hispanics in this same age category. PMID- 15786609 TI - Summary health statistics for the U.S. population: National Health Interview Survey, 2000. AB - OBJECTIVES: This report presents health statistics from the 2000 National Health Interview Survey for the civilian noninstitutionalized population of the United States, classified by age, sex, race and Hispanic or Latino origin, family income, poverty status, education, place of residence, region of residence, and, where appropriate, health insurance coverage. The topics covered are health status and limitations in activities, special education or early intervention services, injuries and poisonings, health care access and utilization, and health insurance coverage. SOURCE OF DATA: The NHIS is a household, multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention, National Center for Health Statistics. Household interviews were completed for 100,618 persons living in 38,633 households, reflecting a household response rate of 89%. SELECTED HIGHLIGHTS: Nearly 7 in 10 persons were in excellent or very good health in 2000, and fewer than 1 in 10 were in fair or poor health. About 31 million people (11%) were limited in their usual activities due to one or more chronic health conditions, and about 3 million people (2%) required the help of another person with activities of daily living such as bathing and dressing. Persons with the least education and the lowest incomes were the most likely to be limited in their ability to work. About 5% of children received special education or early intervention services. The three leading causes of medically attended injury and poisoning episodes were falls, being struck by a person or an object, and transportation. Among people under age 65 years, about 40 million (17%) did not have any health insurance coverage. PMID- 15786610 TI - Summary health statistics for U.S. adults: National Health Interview Survey, 1999. AB - OBJECTIVES: This report presents health statistics from the 1999 National Health Interview Survey (NHIS) for the civilian, noninstitutionalized adult population, classified by sex, age, race and Hispanic origin, poverty status, and region of residence for chronic condition prevalence, health status and limitations in activity, health care access and utilization, health behaviors, and attitudes toward Acquired Immune Deficiency Syndrome (AIDS). Also, health statistics by education, income, health insurance coverage, marital status, and place of residence are presented for health status and limitations in activity, health care access and utilization, health behaviors, and knowledge and attitudes toward AIDS. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized U.S. population. Data are collected during face-to-face interviews with adults present at the time of the interview. The self-reported health information for adults in this report was obtained from one randomly-selected adult per family. HIGHLIGHTS: In 1999, 65% of adults 18 years of age or over reported excellent or very good health. However, 17% of adults less than 65 years of age had no health insurance coverage, and 16% of adults did not have a usual place of medical care. Eleven percent of adults had ever been told by a doctor or health professional that they had heart disease and 19% had been told on two or more visits that they had hypertension. Nearly a quarter of adults were current smokers, while 23% were former smokers. Sixty-one percent did not engage in any leisure-time vigorous physical activity, and, based on estimates of body mass index, 35% were overweight and 21% were obese. PMID- 15786611 TI - [Obstacles in the management of pesticides intoxication in Senegal: about a case of malathion autolysis]. AB - Pesticides are widely used inSenegal as in others africans countries. This situation facilitate occurrence of intoxications, whose management is limited by unavailability of antidotes. We report a case of acute pesticide intoxication in a suicide attempt by a girl of thirteen. The clinical evaluation showed a coma with advanced muscarinic and nicotinic syndrome. The absence of antidotes in the country limited the care management to atropine administration and reanimation measures. The evolution was positive in few days and the patient referred to pediatrics in order to be clinically and psychologically managed. The positive evolution should not hide the necessity for the country to make available all antidotes of toxic substances and to implement Poison Control Centre in order to improve medical management of poisoned people. PMID- 15786612 TI - [Trichobezoard revealed by intestinal perforation. A case report]. AB - This paper presents a case of gastric and ileal trichobezoard on a 12 years-old girl. It was revealeds by ileal perforation which was confirmed by abdominal X ray. Trichobezoard diagnosis was not performed before surgery despite two contradictory/abdominal ultrasound scans. The trichobezoard was removed surgicaly by ileotomy and gastrotomy. The patient had an satisfactory post-operative convalescence after a follow-up of six months. PMID- 15786613 TI - [Fertility, maternal mortality and survival of the children in the medical district of Kolda (Senegal)]. AB - Maternal mortality and lethality in childhood constitute a major problem in Senegal. These two rates are often hight in areas off hight birth rate. In the medical district of Kolda (Senegal) no reliable data on reproductive health exists. We report the results of a preliminary investigation of a community based reproductive project. We've used the sisterhood methods reproductive health's indicators. This cross-sectional study was carried out in a sample of 1287 women in age of procreating. An "aggregative" fertility index of 6.59 was found in the "intervention" area as compare to 5,53 in the control area. The maternal death rate was estimated at 1285 per 100.000 alive births [998-1576]. The infant mortality rate was 83 per 1000. These results justify impremantation of a "vigorous" reproductive health policy in this area. PMID- 15786614 TI - [Study of fear of dental treatments in pediatrics in Senegal]. AB - Fear and anxiety of dental treatment have been recognized as a major source of a problems in pediatric dentistry. They can prevent dental procedure and lead to avoidance an dental phobia. A variaty of studies, using psychometric methods have been carried out worldwide to evaluate these behavious. In Senegal, no study of this nature has been done yet. The aim of this study was to evaluate fear and anxiety related to dental care in senegaleese pediatric children living in Dakar. We have used the children's Fear survey Schedule Dental Sb scale (CFSS-DS) in 415 senegalese children of both sex with a age anking from 7 to 15 years. The results showed a mean score of 33.222 +/- 11.162. Girls revealed a significant higher score (35.122 +/- 11.692) than boys (29.168 +/- 9.704) (p < 0.05). These mean score increase with age in both boys than girl. In addition, more than 43.5% of the children have never consulted a dentist previously and 28% among them showed fear of the dentist. 17% of the children who have dental experiences showed fear of the dentist. This study has showed that senegalease children revealed fear of the dental care even those who have never been to a dentist. It also point out the problem of the origine of dental fear in children. PMID- 15786615 TI - [Tricuspid valve stenosis. A prospective study of 35 cases]. AB - Authors report the results of prospective and longitudinal study. The aims of this study were to evaluate among 35 patients, prevalence, diagnosis and treatment aspects of tricuspid stenosis (TS), as well as evolution and pronostical factors. The prevalence of TS was about 4.2%. The main clinical signs were: dyspnoea (94.2%), jugular veinus pulses (42.8%), superior cave syndrom (68.8%), diastolic rumble (74.3%). ECG showed sinus rhythm (51.4%), a right atrial hypertrophy (48.5%). Echocardiography showed tricuspid leaflets thickened (82.8%), a right atrial hypertrophy (48.5%), a mean gradient between right atrial and right ventricle: 8.6 +/- 3.14 mmHg (65.7%) and mean tricuspid area about 1.41 +/- 0.83 cm2 (continuous equation); about 1.74 +/- 1.29 cm2 (Hatle formula) and 1.11 0.84 cm2 (simplified Hatle formula). Aetiology was only rheumatic fever. After a follow-up of 8.53 +/- 6.06 months, the mortality rate was 28.5%. Complications were irreducible heart failure (24 cases), liver failure (2 cases) and stroke (3 cases). Factors associated with mortality were: severity of tricuspid stenosis and pulmonary hypertension, importance of dyspnea and heart failure (p < 0.041). PMID- 15786616 TI - [The therapeutic itinerapy of epileptics admitted by the Electoencephalographic Laboratory at CHU Dakar]. AB - In Senegal, as in other under developed countries, the delays before seeking medical treatment are long varying from 2 to 12 according to studies. This is related to the fact that in traditional African communities, prejudices and believes delayed medical management which is seeked after failure of traditionnal treatments. The objectives of this study concerning therapeutic itinerary of epileptics in the EEG laboratory at the Neurology Department at CHU in Dakar were to evaluate the average latency of medical consultation and to identify the recourse path by patients receiving antiepileptic treatment. We conducted from March to may 2001, a cross disciplinary study, with a sample size of 212 patients. 79.2% of patients were less than 30 years old. They first seek treatment in hospitals in 36.8%, traditional practionners in 35.8%, health centers in 23%, and private clinics in 4%. The average delay of treatment in modern structures was by 13.4 +/- 4.7 months. The towards the healers was linked to faulty beliefs in 61.8% of the cases, lack of information (22.4%), or a lack of financial means (2.6%). The patients were refeared to traditional healers in 6.7% of the cases for reasons of confidence in traditional medecine. The orientation to neurology clinic was recommended by health personal in 83% of the cases, family (14.6%), or the healer (1.4%). The long and difficult journey of epileptics in Senegal remains marked by exclusion, absence or delay of medical attention. PMID- 15786617 TI - [Bone marrow aspiration in diagnosis of hemopathies in Dakar, Senegal]. AB - The cytological study of bone marrow aspirate, is a useful technique in diagnosis and epidemiological surveillance of hemopathies. Between January 1991 and February 1999, we realized 1000 bone marrow puncture in Dakar hospitals, this study aims to analyze the frequency of diagnosed hemopathies, and to appreciate justification of prescription. Mean age was 31.6 (1 to 88 years). Bone marrow puncture was safe because no incident was observed. Interpretation was possible in 937 cases while the 33 authors was diluted by blood. Bone marrow was normal in 550 cases (57.3%) whereas abnormality was detected in 417 cases (42.7%). The more frequent pathology were quantitative or qualitative defect of bone marrow production with respectively 25.8% and 20.1%, followed by chronic myeloid leukemia (16%), lymphoproliferative disorders (16%), acute leukemias (13.9%), bone marrow metastasis (5.9%), and storage diseases (1.9%). The best justification of the prescription was obtained when done on the basis of hemogram abnormalities or signs in hematopoietic organs with a pathology found in 90% of cases. Prescription on the basis of non hematological signs or to search for metastasis when primitive tumor was not identified were less efficient with respectively 20% and 42% of normal bone marrow. These results emphasize the interest of bone marrow aspiration in epidemiological surveillance of hemopathies, and the advantage to respect the rights indications, in consideration of pain and intolerability of the puncture by patients. PMID- 15786618 TI - [Non iatrogenic primary hypothyrodism in adults at Le Dantec Hospital : clinical features, diagnosis and treatment. Review of 19 cases]. AB - The authors report 19 cases of non iatrogenic primary hypothyroidism in adults at Le Dantec Hospital of Dakar. Those cases had been found during a period of 6 years and half in the internal medicine service. The aim was to study clinical features, diagnosis and outcome of patients after treatment. The mean age of patients was 42.2 years with a sex-ratio of 0.33 M/F. The diagnosis delay was around 6,1 years. All patients presented clinical signs of hypometabolism: physical asthenia (63.15%), frilosity (26.3%), bradycardia (47.3%), constipation (36.8%). The cutaneomucal syndrom was composed by myxoedema (73.6%), macroglossia (26.3%), raucousness of voice (26.3%), alopecia (57.9%). Muscle weakness was found in 2 cases and genital troubles in 3 cases. Five patients presented goiter and 9 others had spontaneous thyroid atrophy. All patients presented a high level of TSH associated with decreased level of T4. Anemia was found in 7 cases and hypercholesterolemia in 13 cases. Treatment was based on substitutive hormonotherapy with L-Thyroxin (75 to 250 microg/day). Evolution was favorable after 10 month mean duration of processing. More alertness is necessary on behalf of the practitioners in front of any sign suggesting hypometabolism to reduce the diagnostic delay and prevent complete form of hypothyroidism that might be complicated, by cardiac involvement in particular. PMID- 15786619 TI - [DRESS syndrome to allopurinol: a case in Dakar]. AB - We report a case of DRESS syndrome also called drug hypersensitivity reaction occurring a 47 years old Senegalese man who has been taking allopurinol for 3 months. That drug was prescribed for peripheric arthralgias associated to a hyperuricemia. He presented a generalised pruritus, cutaneous lesions, fever and facial oedema. On the biological examens, hyperleucocytosis with hypereosinophilia and hyperlymphocytosis associated to the presence of segmented basophiles. In addition, a hepatic cytolysis and cholestasis were documented. Liver ultrasound was normal. The hemocults were negative. These following serologies have been performed and were negative: hepatitis B and C, Epstein Barr virus, cytomegalovirus, syphilis, toxoplasma and parvovirus B19. The anti-nuclear and anti-DNA antibodies were negative. A favourable clinical evolution was remarked after allopurinol treatment withdrawal. A desquamation occurred after 6 days and hemogram turned out to the normal as well as the hepatic tests after 2 weeks. The virologic examens performed 2 months later were unremarkable. This case point out the importance of the early diagnosis and quick withdrawal of the drug in order to prevent serious forms leading to the 10% of death. PMID- 15786620 TI - [Epidemiology and prognosis of caesarean sections in University Hospital of Dakar]. AB - Caesarean section is our first activity in obstetrical surgery. Due to his high frequency and the maternal and foetal prognostic, we have undertaken to do a prospective, longitudinal, exhaustive study concerning all women whose delivery necessitated a caesarean section, between 01 January and 31 December 2001 in Gynaecological and Obstetrical Clinic of University Hospital Center Le Dantec. Our objectives were: describe the epidemiological aspects, evaluate the maternal and foetal prognosis of caesarean sections performed in University Hospital Center. The study revealed that 25.1% of births necessitated a caesarean section. The average age was of 26; the mean parity of 2; the medium height of 161cm. 26.4% of patients were evacuated and coming from the health centers in majority (41.9%) with unmedicalised evacuation in 44.9% of cases. 78.7% of patients have realised at least 3 antenatal consultations. The mains indications were cephalopelvic disproportions (34.9%) and foetal distress (18.9%). The classification per indications group revealed that 37.2% were "prudence caesarean section", 40.1% were "obligator's caesarean section" and 22.7% "necessity's caesarean section". The mean Apgar's score at the birth was 6,5 at first minute. 12 maternal deaths were noted, means 0.8% of global lethality; the morbidity was at 10.5% of cases, represented mainly by postoperative infection (64.3%). The per natal mortality was estimated to 5.9% and neonatal morbidity to 32%. About the quality of caesarean sections, the temporal (24h/24) and financial(kit of caesarean section) accessibilities are more satisfying; otherwise the quality of surgical act and the postoperative follow-up was not optimal. An inflationist advancement is actually noted about the rate of caesarean section, and due to the widening of the indications to the foetal interest. Nevertheless, despite agreed efforts, the quality is still insufficient because the maternal and per natal morbidity and mortality are still high. PMID- 15786621 TI - [Medico-legal conditions of recognition and compensation of occupational asthma in Senegal]. AB - The aims of this study were to appreciate how frequent asthma was named in the legal list of worker's compensation, state hazards and occupations involved and appreciate conditions required for a legal recognition of the disease. The study consisted of a review of the legal list of occupational diseases of all tables where asthma was stated. Medical, technical and administrative criteria allowing compensation were looked for. Asthma was listed in 12 among the 67 tables; causal substances listed were known as sensitizers and included mainly organic and metallic compounds. Numerous occupations were concerned in all primary, secondary and tertiary sectors. Medical conditions refered to "all asthma confirmed by functionally respiratory tests or regenerated by a new exposure". Time length of declaration was seven days. Variety of occupations concerned and numerous substances involved should invite to look for occupations in all patients with asthma. Physicians should be aware of theses legal conditions in order to recognize better occupational asthma and ease it legal compensation. PMID- 15786622 TI - [Ectopia testicular in child: about 3 cases]. AB - The authors report 3 cases of testicular ectopia. Of these 3 cases, two proved tobe cases of penial ectopia whose diagnosis was mentionned before the intervention. One case proved tobe a case of tranverse testicular ectopia diagnosed as an inguino-scrotal hernia. Surgical exploration has always allowed to reach a correct diagnosis. In all three cases the testicle was of normal size and the cord long enough to lower the testicle between the skin an the dartos Surraco's procedure. The authors discuss the pathogenis of this affection, echography contribution in the diagnosis, and the therapeutics aspects. PMID- 15786623 TI - [Child cerebellar meningioma. A case report]. AB - Meningioma is an uncommon tumor of childhood. In this group, supra tentorial forms predominate. We report the case of a 14 years old boy without any past medical history. He presented headaches, vomiting and cerebellar syndrome. CT scan shows cerebellar tumor, enhanced by contrast and surrounded by edema, looking like tuberculoma. After fail of tuberculosis treatment, the boy is operated by suboccipital approach. Histology and immuno histochemical examination show fibroblastic meningioma. Even if it is rare, meningioma can occur in childhood, without neurofibromatosis disease. It can present many points of likeness with tuberculoma and therefore biopsy is mandatory before tuberculosis drugs giving. PMID- 15786624 TI - [Diagnosis delay of multiple myeloma: report of 22 cases in an internal medicine department of Dakar]. AB - Reported is a retrospective study conducted on in patients with multiple myeloma diagnosed from January 1990 to December 2000 in the departement of Internal Medicine I of Dakar University Hospital. Twenty-two patients were included. The mean age was 55 years (range 29-76). The sex ratio (male to female) was 2.6 (they were 16 males and 6 females). The presenting clinical features were dominated by bone manifestations with diffuse pain (41%) followed by pathologic fractures (22.27%) and lumbar pain (13.6%). At the hospital admission the type of clinical manifestations recorded were related to bone (91% general (60%), neurologic (36%) and infection (36%). The biological exams documented an anemia in 20 cases (91%) and increased serum protein in 13 cases (59%) and hypercalcemia in 11 cases (50%) and renal insufficiency in 6 cases (27%). Immunoelectrophoresis had been performed in 6 cases and revaled 4 cases of IgG and 2 cases of light chain myeloma. On the skeletal radiological exams it has been found lytic bone lesions in 66.6 % of cases. diffuse bone rarefaction (38.88%), pathological fractures (38.88%) and vertebral collapses (18.18%). The bone marrow aspiration showed in 17 cases over 19 a medullary dystrophic plasmocytosis. According to the Salmon and Durie classification 68% of the patients were pointed in stage III. Comparing our results to the series, we figure out our patients younger at the time of the diagnosis and an important diagnosis delay as previously suggested in the African litterature. This is confirmed by the fact that 68% of patients were one Salmon and Durie stage III. the frequency of severe anemia and pathological fractures We invite practitionners to conduct systematic exploration of mild symptoms like lumbar pain. PMID- 15786625 TI - [Hepatitis B infection in infantile population of Senegal]. AB - Hepatitis B is endemic in Senegal. According to many data, the prevalence of this infection in adult population is up 85%. Young children are victims of the intensive circulation of this virus. Indeed, the risk of becoming chronic carrier which can further lead to Hepatocellular Carcinoma, is related to the age at which the infection had been contracted. In this study, we investigated the prevalence of HBV markers in children less than 5 years living in two regions of Senegal: Dakar and Thies. Using specific Elisa methods, HBV markers were determined in 2962 sera of newborns and children: AgHBs, antiHBc, antiHBs for all children; AgHBe and IgM antiHBc for AgHBs carriers. HBV markers were detected in 59.38% of sera; in 39.26% of the samples only one marker was detected: AgHBs (3.18%), antiHBc (36.08%), antiHBs (0.49%). In 20.12% of samples, markers were associated. So, the global prevalence of the infection is 59.38% with the predominance of chronic forms compared to acute one. The rate of chronic carriage is independent of the sex but is influenced by age and geographic area. The post- infection immunization rate is 7.11%. In Senegal Hepatitis B is contracted at the early childhood. The high prevalence of this infection in children under five years emphasize the need for a spread vaccination in this age bracket, since we well know that the risk for them to become chronic carriers is high. PMID- 15786626 TI - [Epidemiology, prognosis and prevention of choriocarcinoma in University Hospital of Dakar]. AB - The objective of this study was to define epidemiological profile of choriocarcinoma in Senegal, to evaluate its prognosis and to submit a better strategy of prevention. It's a retrospective cases-control study; all choriocarcinoma diagnosed from January 1st to December 31st 2000 at Dakar University Teaching Hospital. The control were chosen among patients who had molar abortion during the same time than the cases and did not developed the pathology after at least 12 months of medical follow up. The stability of association is checked by calculating 95 confidence interval by Miettinen method, and using Chi 2 test of Pearson with a p value of 5% or Fischer test; the prognosis survival factors was compared using logrank test. Among 1098 patients with medical care follow up of post molar abortion, 61 choriocarcinoma were diagnosed with an incidence of 5.5%. The epidemiological profile is a great multipare 40 years old or more with low socio-economical status, 51% of diagnosed cases was in metastasis stage. The prognosis was marked by a complete remission in 37.7% and by global lethality in 49.2%; the average survival was about 48 months. The risk factors of choriocarcinoma among patients who had a molar abortion are represented by an age superior or equal 40 years old, high multipare and preservation of the uterus after molar abortion. To improve the prognosis we recommend to widen preventive hysterectomy indications after molar abortion among patients with risk factors of choriocarcinoma. PMID- 15786627 TI - Summary health statistics for U.S. children: National Health Interview Survey, 2000. AB - OBJECTIVES: This report presents statistics from the 2000 National Health Interview Survey on selected health measures for children under 18 years of age, classified by sex, age, race, Hispanic origin, family structure, parent's education, family income, poverty status, health insurance coverage, residence, region, and health status. The topics covered are asthma, allergies, learning disability, Attention Deficit Hyperactivity Disorder (ADHD), prescription medication, respondent-assessed health status, school-loss days, usual place of health care, time since last contact with a health care professional, unmet dental need, time since last dental contact, and selected measures of health care access. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized population of the United States. Data are collected during face-to-face interviews with adults present at the interview. Information about children is collected for one randomly selected child per family in face-to-face interviews with an adult proxy respondent familiar with the child's health. SELECTED HIGHLIGHTS: In 2000 most U.S. children under 18 years of age enjoyed excellent or very good health (83%). However, 12% had no health insurance coverage, and 6% had no usual place of health care. Twelve percent of children had ever been diagnosed with asthma. Eight percent of children 3-17 years of age had a learning disability, and 7% of children in this age group had ADHD. Lastly, 11% of children in single-mother families had two or more visits to an emergency room in the past year compared with 6% in two-parent families. PMID- 15786628 TI - Clinical perspectives in type 2 diabetes care: optimizing glycemic control with insulin therapy. PMID- 15786629 TI - Deaths: leading causes for 2002. AB - OBJECTIVES: This report presents final 2002 data on the 10 leading causes of death in the United States by age, race, sex, and Hispanic origin. Leading causes of infant, neonatal, and postneonatal death are also presented. This report supplements the annual report of final mortality statistics. METHODS: Data in this report are based on information from all death certificates filed in the 50 States and the District of Columbia in 2002. Causes of death classified by the International Classification of Diseases, Tenth Revision (ICD-10) are ranked according to the number of deaths assigned to rankable causes. RESULTS: In 2002, the 10 leading causes of death were (in rank order) Diseases of heart; Malignant neoplasms; Cerebrovascular diseases; Chronic lower respiratory diseases; Accidents (unintentional injuries); Diabetes mellitus; Influenza and pneumonia; Alzheimer's disease; Nephritis, nephrotic syndrome and nephrosis; and Septicemia and accounted for about 79 percent of all deaths occurring in the United States. Differences in the rankings are evident by age, sex, race, and Hispanic origin. Leading causes of infant death for 2002 were (in rank order) Congenital malformations, deformations and chromosomal abnormalities; Disorders related to short gestation and low birthweight, not elsewhere classified; Sudden infant death syndrome; Newborn affected by maternal complications of pregnancy; Newborn affected by complications of placenta, cord and membranes; Accidents (unintentional injuries); Respiratory distress of newborn; Bacterial sepsis of newborn; Diseases of the circulatory system; and Intrauterine hypoxia and birth asphyxia. Important variation in the leading causes of infant death is noted for the neonatal and postneonatal periods. PMID- 15786631 TI - Myths about the brain. PMID- 15786630 TI - A matter of privacy. PMID- 15786632 TI - Luxury primary care...a new idea. PMID- 15786633 TI - Glaucoma without cupping. PMID- 15786634 TI - Use of the artificial iris implant in patients with aniridia. AB - BACKGROUND: Aniridia (or partial aniridia) often occurs as a result of a penetrating ocular trauma. This condition may cause symptoms including glare, light sensitivity, reduced vision, and asymmetric appearance. Options for these patients include specialty contact lenses, corneal tattooing, and artificial iris implants. This article details six patients who experienced penetrating trauma with significant iris tissue loss and who chose to have an artificial iris implant. CASE REPORTS: Six patients with traumatic aniridia were enrolled in a single-site, single-surgeon study. They were followed for one year postoperatively. All six were complex cases that had multiple eye problems, such as aphakia, corneal scarring, corneal graft rejection, and retinal detachment. All participants were male. All patients experienced decrease in glare and light sensitivity following artificial iris implant surgery. Two experienced improved best-corrected vision (BCVA). All six patients felt the cosmetic appearance of their affected eye improved. CONCLUSIONS: The artificial iris device provides an effective means of treating traumatic aniridia. Optometrists often work with patients who have experienced a penetrating ocular trauma, with complete or partial aniridia as a complication. In cases in which nonsurgical means are not effective in returning the patient to comfort and productivity, an artificial iris implant should be considered. PMID- 15786635 TI - Tuberculum sellae meningioma associated with lymphomatoid papulosis. AB - BACKGROUND: Meningiomas involving the tuberculum sellae and planum sphenoidale typically cause visual loss by affecting the optic nerve and anterior chiasm. We present a case of a tuberculum sellae meningioma concurrent with lymphomatoid papulosis, a T-cell lymphomatoid skin disorder. CASE REPORT: A 50-year-old black woman came to the eye clinic with a symptom of decreased vision in the right eye for one week, especially when reading. Best-corrected vision was counting fingers at two feet O.D. and 20/40 O.S. She was seen seven months earlier, with 20/25 O.D. and O.S. Dilated fundus examination revealed bilateral optic atrophy, O.D. greater than O.S. MRI revealed a meningioma arising from the tuberculum sellae and planum sphenoidale region. The meningioma was partially resected nine days later Visual field performed 18 days postoperatively demonstrated an overall depression in the right eye and no defect in the left eye. Best-corrected vision improved to 20/150 O.D. and 20/30 O.S. CONCLUSION: Meningiomas are generally benign tumors that can cause symptoms if vital structures are compromised. Altered visual function and optic atrophy may be the only presentation of intracranial and orbital tumors. Intracranial meningiomas may be associated with systemic conditions that can infiltrate the optic nerve, including lymphocytic disorders. Lymphomatoid papulosis (LyP) is a rare cutaneous disorder involving infiltrating clonal T-cells that has been associated with disseminated lymphomatic skin tumors. It is unclear if there is a direct association between LyP and meningiomas. PMID- 15786636 TI - Acinetobacter as a causative agent in preseptal cellulitis. AB - BACKGROUND: The bacterial genus Acinetobacter, once viewed as inconsequential, has been increasingly implicated as a significant pathogen. Acinetobacter is becoming progressively more resistant to older and newer-generation antibiotics. The primary species encountered in infections is Acinetobacter baumannii. CASE REPORT: A 39-year-old woman came in for treatment of a preseptal cellulitis and conjunctivitis. She was referred to W.W. Hastings Optometry Clinic eleven days after first being diagnosed with a hordeolum that developed into a preseptal cellulitis, despite the use of oral cephalexin anazithromycin. After being referred, the patient was placed on oral regimen of gatifloxacin and topical ciprofloxacin. The patient gradually improved over the next 10 days, with full resolution occurring 21 days after the first presentation. Culturing revealed the offending organism to be Acinetobacter. DISCUSSION: Most ocular infections are still caused by common organisms such as Staphylococcus. However, atypical bacteria such as Acinetobacter are responsible for a minority of infections. Careful consideration of patient history and clinical signs must be made to differentiate preseptal cellulitis and orbital cellulitis. CONCLUSION: This case emphasizes that clinicians need to be aware of emerging and resistant bacteria in ocular infections. Infections that do not respond to traditional therapy should be treated aggressively with the latest-generation antibiotics. Ocular infections that do not resolve in a timely manner should be cultured for the offending organism and these organisms should be tested for susceptibility to anti microbial drugs. PMID- 15786637 TI - Multiple calcified deposits along the eyelid margins secondary to chronic renal failure and hyperparathyroidism. AB - BACKGROUND: Corneal and conjunctival calcification have been associated with chronic renal failure and hyperparathyroidism. Corneal alterations include band keratopathy and/or limbal involvement similar to limbal girdle of Vogt. Conjunctival lesions resemble pingueculae. Lid lesions, which include calcified deposits adjacent to the meibomian gland orifices, are much less common. CASE REPORT: A 52-year-old woman came to SUNY State College of Optometry clinic with a medical history of gestational hypertension, which led to chronic kidney failure and secondary hyperparathyroidism. Slit-lamp examination revealed multiple small, white, round calcium deposits, which were found just posterior to the Meibomian gland orifices, along the superior and inferior lid margins. CONCLUSION: Ocular calcification has been observed in patients with certain systemic diseases such as chronic renal failure, hyperparathyroidism, hypophosphonemia, sarcoidosis, hypervitaminosis D, and metastatic neoplasia. We present the third report of eyelid calcification associated with a systemic disease. PMID- 15786639 TI - What is your management personality? PMID- 15786638 TI - The IOL horizon: accommodative intraocular lenses. AB - BACKGROUND: For more than half a century, intraocular lenses have been used effectively to correct vision after cataract extraction. New developments in intraocular lens design have led to the creation of lenses that may have a significant role in the restoration of accommodation as well. PURPOSE: Accommodative lens technology is constantly evolving. Many different lens designs are currently under development. It is important for optometrists to be familiar with this technology and the various lenses being developed, as accommodative intraocular lenses stand to become the mainstay for cataract surgery, as well as for the surgical treatment of presbyopia. METHOD: The literature is reviewed in order to summarize the developments in accommodative lens technology. PMID- 15786640 TI - Physical safeguards for the optometric practice. PMID- 15786641 TI - CSI: common stategies for the infant. PMID- 15786642 TI - Employees and patient confidentiality. PMID- 15786643 TI - [All is endocrinology]. PMID- 15786644 TI - [Genetics of multiple endocrine neoplasies: clinical implications]. AB - The multiple endocrine neoplasia (MEN) syndromes are hereditary monogenic diseases that are transmitted as autosomal dominant traits, and are characterized by the development of tumors and hyperplasias in several endocrine organs. The causative genes of the 2 principal forms of MEN have been recently identified; a protooncogene for MEN2 (the RET gene) and a tumor suppressor gene for MEN1 (the MEN1 gene). Correlations between phenotype and genotype were described in the case of RET mutations that could help in defining the screening methods and the preferable age of prophylactic thyroidectomy. No correlations were established between the mutations of the MEN1 gene and the phenotype of patients suffering from MEN1. We present here a synopsis of the recent results of the genetics of MEN syndromes underlining their clinical implications. PMID- 15786645 TI - [Focus on dehydroepiandrosterone (DHEA)]. AB - Secreted by the adrenal cortex, DHEA exerts its action either indirectly in peripheral tissues after its conversion to androgens and estrogens, or directly as a neurosteroid through the interaction with neuronal receptors. Analyses of randomized studies show that treatment with DHEA improves well-being and fatigue in patients with adrenal insufficiency and reduces disease activity in women with systemic lupus erythematosus. Interesting results have also been observed in the treatment of depressive disorders, but these studies require confirmation. In contrast, there is neither justification for DHEA supplementation in healthy elderly subjects nor clear evidence for beneficial effects of DHEA on muscle function, bone metabolism or cognition. Finally, there is no guarantee with regard to the quality of the product or its safety during long term use. PMID- 15786646 TI - [Hirsutism]. AB - The presence of terminal hair with a male pattern of distribution in a women is called hirsutism, and is either due to increased androgen production or to increased sensitivity of the hair follicle to androgens. In more than 95% of cases, it is a benign condition, generally a polycystic ovarian syndrome. When there is no evidence of oligo-anovulation and androgen levels are normal, the diagnosis of idiopathic hirsutism is given. In both situations, treatment is threefold: 1) inhibition of androgen production; 2) inhibition of androgen action and 3) a local treatment. The brisk apparition and extension of hirsutism, together with signs of virilisation and high levels of circulating androgens, suggest the presence of a tumor, extremely rare, but almost always malignant. PMID- 15786647 TI - [Growth hormone treatment: pediatric to adult clinic transition]. AB - The diagnosis of GH deficiency is difficult to establish: clinical, radiological and hormonal data are combined to suspect the disease. GH stimulation tests are an essential part of the evaluation, although the cut-off values are determined arbitrarily. There are different stimulation tests. Their use depends on the patient's age. Once the diagnosis is ascertained, the treatment is started and maintened until the end of statural growth. The persistence of GH deficiency needs to be confirmed during the transition phase. If required, GH treatment can be continued until the achievement of peak bone mass. Thereafter the benefit of continuing GH treatment are mainly related to the quality of life. The long term effects on cardiovascular morbidity/mortality are not demonstrated. PMID- 15786648 TI - [Thyroiditis: an approach for the practitioner]. AB - Thyroiditis represent commonly encountered thyroid disorders in clinical medicine. Autoimmune thyroiditis is frequent with various forms: Hashimoto's thyroiditis, subacute thyroiditis with lymphocytic and post-partum variants. In the absence of any changes in volume and function, Hashimoto's thyroiditis can be seen in a setting of a thyroid nodule, elevation of thyroid autoantibodies and by the pathologist in surgical or cytological materials. Presenting with fever, cervical pain and subtle signs of hyperthyroidism, subacute thyroiditis (De Quervain) represents often a challenge for an early recognition and an appropriate treatment. Acute thyroiditis and Riedel's thyroiditis are very uncommon, with cervical mass and pain. PMID- 15786649 TI - [Medical treatment of prolactinoma. Quo usque tandem..]. AB - Dopamine agonists are the first choice therapy for prolactinoma. They decrease tumor secretion and volume, restore the gonadotropic axis, preserve the remaining pituitary functions, and correct the ophthalmologic symptoms. They do not cause hypopituitarism, contrary to surgery or radiotherapy. The latter are reserved for the rare patients who display an inappropriate response to dopamine agonists. It is important to determine the circumstances under which remission is observed after prolonged drug therapy. Recent data indicate significant percent remission in patients with macro- or microprolactinoma. However, the data do not allow to determine how therapy should be performed to better reach that goal, nor to identify predictive factors of this favorable outcome. PMID- 15786650 TI - [After your heart arrest, would you like to test a medicinal elixir?]. AB - So far, cardiac arrest is still associated with high mortality or severe neurological disability in survivors. At the tissue level, cardiac arrest results into an acute condition of generalized hypoxia. A better understanding of the pathophysiology of ischemia-reperfusion and of the inflammatory response that develops after cardiac arrest could help to design novel therapeutic strategies in the future. It seems unlikely that a single drug, acting as a <>, might be able to improve survival or neurological prognosis. Lessons learned from pathophysiological mechanisms rather indicate that combined therapies, involving thrombolysis, neuroprotective agents, antioxidants and anti inflammatory molecules, together with temperature cooling, might represent helpful strategies to improve patient's outcome after cardiac arrest. PMID- 15786651 TI - [The first worldwide double hand transplant, 5 years later]. PMID- 15786652 TI - [Biomedicine and procreation: what will the future hold?]. PMID- 15786653 TI - [The Pope and the enlightenment of fermented papaya]. PMID- 15786654 TI - [Confronting complexity]. PMID- 15786655 TI - Herpes simplex virus infection: part I--Biology, clinical presentation and latency. AB - Oro-facial manifestations of herpes simplex virus (HSV) infections are very common, and include primary herpetic gingivo-stomatitis, recurrent herpes labialis and recurrent intra-oral herpes. Recent research in molecular biology has advanced our knowledge of the HSV pathogenesis and behavior. Understanding the exact mechanism of HSV latency and reactivation enables improvement of drug therapy and prevention strategies of HSV infections. The aim of this review is to update the recent development in the biological and clinical research related to HSV infection, focusing on oral and perioral lesions. PMID- 15786656 TI - [Implants placement using a novel computerized tactile imaging navigation system]. AB - An implant locating system has been developed. The system contains a stent that maps the jaw bone at the implantation site through the tissue, sends a signal to a computer with a CT scan of the jaw on it, and superimposes the position of the stent on the jaw so that a dental surgeon could angle the implant for placement without requiring to raise a flap to expose the bone. Using a novel tactile technology the system allows safe, accurate, and simple implant placement and design. The ILS software allows: 1. Importing of CT data. 2. Marking a dental arc on the computerized jaw image. 3. Planning of implant location and position on a 3-D view. 4. Affixation of the ILS to the jaw, followed by registration of the stent. 5. Navigated osteotomy and implant placement. PMID- 15786657 TI - [Oral hygiene, caries status and bad breath among young Israeli recruits]. AB - The purpose of the present study was to evaluate the extent of self-reported bad breath in an Israeli population of young army recruits and to assess its relationship with other self-reported parameters, as well as general dental status. Self-reported parameters included smoking status, bad taste, gingival bleeding and the presence of tonsilloliths. The study comprised 426 young adults recruits (ages 18-19), almost all males (95%), all of whom agreed to answer a questionnaire. All participants underwent a dental screening and were divided into three groups regarding to their dental status (do not need treatment, need moderate treatment, need extensive treatment). Statistical analysis included Pearson's chi square test of association using BMDP statistical software. Among the recruits, 142 (33%) were active smokers. Thirty-five participants (8.2%) reported bad breath as well as bad taste. Twenty-seven (6.3%) reported being told that they had bad breath. Tonsilloliths were reported by 31 participants (7.3%) and gingival bleeding by 80 (18.8%). Self-reported bad breath was positively associated with bad taste, gingival bleeding, the presence of tonsilloliths and general dental status (p<0.05). The study indicates that self reports of bad breath are associated by objective factors (e.g. dental status, tonsilloliths) as well as subjective parameters (bad taste). PMID- 15786658 TI - [Endodontic surgery (apicoectomy)--success rate of more than 90% using dental operating microscope and ultrasonic tips]. AB - The main cause of root canal treatment failure is presence of bacteria in the root canal. Surgical endodontic retreatment is indicated as a valuable treatment choice for failed endodontic treatment. The aim of the article is to describe the advantages of a new technique for apicoectomy using dental operating microscope and ultrasonic tips in comparison to the traditional technique. Precise root end resection with minimal or no bevel and accurate preparation of the root end cavity to the depth of 3-4 mm allows minimal bone removal and reduces procedural accidents such as perforation of the lingual canal wall. Zink oxide eugenol based materials allow sealing of the retrograde preparation adequately to prevent bacteria and toxins to penetrate the periradicular tissues. Literature review demonstrates success rate to above 90% when employing dental operating microscope and ultrasonic tips for retrograde cavity preparation. PMID- 15786659 TI - [Radiographic identification of dental implants in Israel]. AB - Not infrequently a patient will attend seeking treatment for replacement of a lost abutment and crown from an implant placed by another dentist and no dental records are available. Many different root form implants of various designs, dimensions, surfaces, and connectors are available today posing a problem for the restorative dentist who must choose prosthetic parts that fit the implant or for a forensic dentist attempting to identify a victim. Implants installed in bone may be identified only by radiographs and knowledge of implant morphology. Four articles (1-4) were published between 1992-2002 dealing with the radiographic identification of implants. Since then many new implants have appeared on the market. Implants made in Israel do not appear at all on any of the lists. The object of this article is to provide radiographic pictures of implants available in Israel today together with a description of their radiographic features to enable Israeli dentists to identify implants where no dental records are available. PMID- 15786660 TI - [Image guided dental implantology]. AB - Dental implants insertion is a predictable surgical procedure with very high success rates. An optimal implants placement requires excellent surgical skills and good prosthetic perception. Performing an inaccurate implantation can lead to irreversible surgical damage on the one hand or a prosthetic failure on the other hand. Planning software provide the surgeon with good planning tool; existing navigation systems allow for translating them into performance by semi-active or passive guidance. The later allows for flexibility in the implant location during the operation and real-time tracking of drill position. All of these tools are helpful in avoiding damage to anatomical structures by performing the implantation in close relation to the CT scan. But the solutions that provide most possible advantages requires CT with special markers, long and expansive preoperative preparations and most of all a very high initial cost. These, in addition to a very long learning curve are the reason for these systems not to become a popular working tool. The most important challenges of the next generation systems in dental implants navigation are lower price, smaller size, good performance and reliability and ease of use. This kind of image guided system should allow for preplanning of implants locations, and guided insertion by minimal invasive procedure. PMID- 15786661 TI - [Specifying a field of practice in dentistry when lacking specialty]. PMID- 15786662 TI - Preterm low birthweight (PLBW) and maternal periodontal disease: a debate in the literature. PMID- 15786663 TI - A golden opportunity. PMID- 15786664 TI - Pain relief. PMID- 15786665 TI - The missing link. PMID- 15786666 TI - Ancient exercise for modern rehab. PMID- 15786667 TI - To stand or not to stand. PMID- 15786668 TI - Rolling stock. PMID- 15786669 TI - Sit on it. PMID- 15786670 TI - Saving the hospital-within-the hospital. PMID- 15786671 TI - A more than modest proposal. PMID- 15786672 TI - Stroke prevention in atrial fibrillation: are we following the guidelines? AB - BACKGROUND: Warfarin therapy substantially reduces stroke in atrial fibrillation (AF), yet medical literature reports it is only prescribed in 15-60% of eligible patients. No current national benchmarks for warfarin use in AF patients exist, and it is unclear whether the reported poor compliance represents current rates within primary care practices. The primary study objective was to measure the rate of warfarin use in eligible, high-risk AF patients in a large southeastern group family practice. Secondary objectives were to report the demographics, stroke-risk profiles, contraindications, and reasons for discontinuation of warfarin therapy METHODS: A retrospective chart review was performed on all active patients with documented AF in a large southeastern group family practice/residency between July 1, 2000 and June 30, 2002. Data was abstracted on warfarin use, contraindications, stroke risk, and reasons for discontinuation. RESULTS: Four hundred ninety-one (491) patients were identified from the electronic billing system as potential study subjects. Two hundred eighty-three (283) patients met study criteria, with 210 patients considered to be at high risk of stroke without contraindications to warfarin therapy. Ninety-four percent (198/210) of these patients were prescribed warfarin during the study period, and 87% (172/198) continued warfarin throughout the study period. CONCLUSION: Family physicians in this practice prescribe warfarin in AF more frequently than published rates demonstrating that high rates of physician adherence to standards are achievable in primary care. Most patients in this setting were considered high-risk for stroke. PMID- 15786673 TI - Alzheimer's disease and the family caregiver: the cost and who pays? PMID- 15786674 TI - The Alzheimer's Association national policy agenda. PMID- 15786675 TI - Alzheimer's and the caregiver [interviewed by Donald L. Madison]. PMID- 15786676 TI - Grieving the living! PMID- 15786677 TI - Family care and Alzheimer's disease: what do we know? What can we do? PMID- 15786678 TI - North Carolina's support of family caregivers. PMID- 15786679 TI - Dementia-specific respite: the key to effective caregiver support. PMID- 15786680 TI - Managing the patient with dementia in primary care. PMID- 15786681 TI - Alzheimer's disease: a nursing home perspective. PMID- 15786682 TI - Preparing families to make informed decisions about long-term care. PMID- 15786683 TI - Dementia care: an outpatient, community-based, multi-disciplinary approach. PMID- 15786684 TI - Alzheimer's disease, family caregivers, and Faith in Action in North Carolina. PMID- 15786685 TI - Dementia-responsive acute care at Mission Hospitals in Asheville, North Carolina. PMID- 15786686 TI - Deaths and hospitalizations from Alzheimer's disease in North Carolina. PMID- 15786687 TI - New York Times. PMID- 15786688 TI - Quality of care and performance improvement. PMID- 15786689 TI - Eliminating racial and ethnic health diparities. PMID- 15786690 TI - Eliminating racial and ethnic health diparities. PMID- 15786691 TI - Biotechnology and the birth of a third culture. AB - Biotechnology represents such an important challenge for present day culture that one can speak of a biotechnological revolution in many other scientific fields as well, such as biology, clinical medicine, pharmacology, and genetic engineering. It also significantly affects political and economic choices to such a degree that they call for a new kind of attention from jurisprudence which has to regulate an ever changing world. Many important queries arise particularly at a bioethical level, issues that will also affect future generations. Scientific progress has unexpectedly widened the biological knowledge of human kind. Thanks to the contribution of continuously more refined and advanced technology, it has nurtured the hope of solving all problems and of overcoming all limits. The scientist's intellectual curiosity, encompassing these new resources, is spurred on by the desire for knowledge and understanding. However sometimes he loses sight of the repercussions and of the possible uses his achievements may have. Only a profound personal education, integrated with the scientist's technical and scientific expertise, will allow science to knock down some barriers, advancing constantly but without losing respect for man's dignity. However the separation between scientific and ethical expertise can only raise new barriers and create limits to the freedom of science which will appear just restrictive, while a kind of medieval obscurantism will be opposed to ethical rigour. PMID- 15786692 TI - On the holistic approach in cancer biology: tumor necrosis factor, colon cancer cells, chaos theory and complexity. AB - TNFalpha plays a role in the pathogenesis of septic shock, inflammatory diseases, autoimmune diseases, graft rejection reaction, acute, and chronic respiratory inefficiency among others. Its activity depends on the type of target cells and different regulating factors, but the effect of biological activity is conditioned by specific receptors such as p55 (type I, TNF R55) and p75 (type II, TNF R75). The aim of the study was to answer the following questions: 1) Is it possible to apply elements of non-linear dynamics to assess the level of expression of TNF, TNFRI, TNFRII genes in tumor cells, pathologically unchanged tissue and metastatically changed lymph nodes? 2) Is theoretically anticipated variability of cytokine and its receptors in colorectal carcinoma cells and the immediate vicinity justified in the developed mathematical model? The research material--specimens taken from tumor, unchanged tissue and metastatic lymph nodes -were histopathologically and molecularly analysed. Results of the molecular research were used to develop a mathematical model using the basic studies on the theory of chaos and biological system modelling. PMID- 15786693 TI - Preventing sexual transmission of HIV: anti-HIV bioregulatory and homeostatic components of commercial sexual lubricants. AB - Certain safe over-the-counter (OTC) sexual lubricants such as Astroglide, KY Liquid, Replens, Vagisil, ViAmor, and Wet Stuff inhibit both cell-free HIV and the production of HIV by infected leukocytes in vitro even in the presence of seminal fluid. To identify which components of the lubricants were active against HIV, we tested five components (glycerin, methylparaben, propylparaben, polyquaternium-32, and propylene glycol). The paraben preservatives and propylene glycol in the lubricants did not inhibit HIV, while the common natural homeostatic metabolite, glycerin, and the thickener polyquaternium-32 did strongly inactivate infectious HIV and HIV-infected leukocytes. Activity against HIV and HIV-infected cells by glycerin was stable through 24 hours at 37 degrees C. Glycerin and polyquaternium-32 were active at minimum concentrations of approximately 2% and 0.01%, respectively--well within the highest FDA safety guidelines. Both active components disrupted infected leukocytes within 5 minutes which resulted in inhibition of infectious HIV production by infected leukocytes of greater than 25 to 100-fold. These components do not disrupt vaginal epithelial cells in vivo. These components also rapidly inactivate cell-free HIV by 10- to 30-fold. Thus, we may conclude that the active components of the OTC lubricants are glycerin and polyquaternium-32. Using these components, OTC sexual lubricants could be reformulated to optimize their anti-HIV activity. Furthermore, clinical trials of these lubricants and components seem to be indicated because of their FDA safety level, wide availability, and low cost. PMID- 15786694 TI - Biology and novel therapeutics for neuroendocrine tumors of the lung. AB - Neuroendocrine (NE) tumors of the lung are a special class of tumors that include large cell neuroendocrine carcinoma (LCNEC), typical carcinoid (TC) tumor, atypical carcinoid (AC) tumor, and small cell lung cancer (SCLC). Even though they all have the same NE phenotype, these tumors can differ in their pathological characteristics, immunohistochemical patterns, molecular and cellular biology, clinical characteristics, response to various therapeutic modalities, and finally the ability to be molecularly targeted by novel therapeutics. In this review article, we will summarize the various characteristics of these specialized NE tumors, with particular emphasis on the biology with the potential for novel targeted therapies. As an example, SCLC is characterized by overexpression of receptor tyrosine kinases such as c-Kit, c-MET and Ret, and these can be targeted with small molecule inhibitors and various antibodies. Many of NE tumors are quite aggressive and arriving at targeted therapies would be a useful venue to pursue for a potential cure. PMID- 15786695 TI - Biological response modifiers of cancer-related neuroendocrine disorders: efficacy of the long-term dopaminergic agonist cabergoline in the treatment of breast cancer-induced hyperprolactinemia. AB - The evaluation of the biological status of cancer patients should not be limited only to investigation of immune reactivity, but should also include analysis of the endocrine condition, namely concerning those hormones which have appeared to be tumor growth factors, such as prolactin (PRL) for breast and prostate carcinomas. This statement is justified by the fact that the evidence of abnormally high serum concentrations of PRL has been proven to be associated with poor prognosis in breast and prostate cancer patients. Moreover, since hyperprolactinemia negatively influences the efficacy of anticancer therapies in breast cancer, it could be fundamental to achieve a normalization of PRL levels by long-acting dopaminergic agents, such as cabergoline. On this basis, a study was planned to evaluate the effect of cabergoline on PRL levels in hyperprolactinemic metastatic breast cancer subjects. The study included 20 hyperprolactinemic metastatic breast cancer subjects, who were randomized to receive no therapy or cabergoline at 0.5 mg/week orally for 4 consecutive weeks. Cabergoline therapy induced a normalization in all patients, whereas no spontaneous normalization of PRL levels occured in the control group. These results show that a weekly oral administration of the long-acting dopaminergic agent cabergoline is a well tolerated and effective treatment of metastatic breast cancer-related hyperprolactinemia. The possible prognostic impact of PRL normalization needs to be established by successive studies. PMID- 15786696 TI - Simvastatin decreases concentration of interleukin-2 in hypercholesterolemic patients after treatment for 12 weeks. AB - Statins, HMG-CoA reductase inhibitors are drugs with a potent lipid-lowering effect. They are also able to inhibit proliferation of smooth muscle cells, T lymphocytes, to restore endothelial activity and to inhibit inflammatory responses. These effects have been called the pleiotropic effect of statins. Statins have demonstrated contrast to the inflammatory activity of macrophages. The aim of the study was to assess the influence of simvastatin on serum levels of proinflammatory cytokines such as IL-2 and TNFalpha in hypercholesterolemic patients. METHODS: In 58 non-smoking men with total cholesterol (TC) >7.8 mmol/L, LDL-cholesterol>5.5 mmol/L and fasting triglycerides<4.6 mmol/L serum IL-2 and TNFalpha were determined at the beginning of the study, after 3 months diet and after 3 months of simvastatin therapy (20 mg/day). The control group was composed of 10 healthy volunteers with correct lipid values: TC<5.2 mmol/L, LDL cholesterol <2.3 mmol/L, HDL-cholesterol >1.5 mmol/L and triglycerides<2.3 mmol/L. RESULTS: There were significant reductions in IL-2 concentration after 3 months diet (p=0.0059) and significant (p=0.0003) decrease of IL-2 after 3 months of simvastatin therapy. Meanwhile we observed a significant decrease of TNFalpha concentration after 3 months diet (p=0.0001) and no significant decrease after 3 months of simvastatin therapy. PMID- 15786697 TI - Impact of standardization on clinical cell analysis by flow cytometry. AB - The evolution of flow cytometry from a research tool to a pivotal technology for clinical diagnostic purposes has required significant efforts to standardize methods. The great advantage of flow cytometry is that it's applications are highly amenable to standardization. Here, we review the efforts that have been made for flow cytometric applications in four major fields of clinical cell analysis: CD4+ T-cell enumeration, CD34+ hematopoietic stem and progenitor cell enumeration, screening for the HLA-B27 antigen and leukemia/lymphoma immunophenotyping. These standardization efforts have been parallelled by the establishment of external quality assessment (EQA) schemes in many countries worldwide. The goal of these EQA exercises has been primarily educa-tional, but their results will increasingly serve as a basis for laboratory accreditation. This important development requires that the EQA schemes, in particular the quality of the distributed samples and the procedures for evaluating the results, meet the highest standards. PMID- 15786699 TI - Pro-T ALL: immunophenotypical analyses. AB - Pro-T acute lymphoblastic leukemias (ALL), also called T-I ALL, are extremely rare diseases where cells at the earliest defined stage of T-lineage maturation proliferate in the bone marrow. They are characterized by the absence of all T lineage differentiation markers except intracytoplasmic CD3 and surface CD7. They may express markers of immaturity and a number of myeloid lineage markers, without reaching the score defining biphenotypic acute leukemia (BAL). A literature search was performed on Medline and the few publications devoted to this disorder are reviewed in this manuscript. Attempts at understanding the mechanisms leading to the development of T-I ALL, based mostly on animal experiments are further presented. PMID- 15786698 TI - Three-color flowcytometric analysis of mature and immature hematological malignancies. A guideline of the Dutch Foundation for Immunophenotyping of Hematological Malignancies (SIHON). AB - Multiparameter flowcytometry offers an insight into differentiation pathways, maturation stages and abnormal features of cell (sub)populations thus helping to establish and classify hematological malignancies. The Dutch Foundation for Immunophenotyping of Hematological Malignancies (SIHON) has formulated a guideline for a rapid screening followed by confirmation and classification in a standardized way. For this aim seven carefully composed monoclonal antibody combinations are elucidated for screening the test sample in a first phase. In this phase a relative frequency distribution of the cells will be established and a decision will be made about abnormal cells present, as well as their mature or immature state and the cell lineage they belong to. In a second phase, panels with cell lineage dependent monoclonal antibody combinations may be used to confirm and classify the abnormal cell population indicated in phase 1, as well as to establish the presence or absence of an abberant immunophenotype. PMID- 15786700 TI - Reactive phenotypes after acute and chronic NK-cell activation. AB - Several phenotypic changes have been shown to occur after NK-cell stimulation, involving molecules that have been proved to regulate NK-cell migration into tissues and NK-cell activation and proliferation as well as target cell recognition and killing. Here, we review the reactive phenotypes observed in vivo after acute and chronic NK-cell activation. PMID- 15786701 TI - Myeloid cell-associated lysosomal proteins as flow cytometry markers for leukocyte lineage classification. AB - Lysosomal proteins including myeloperoxidase (MPO), lysozyme (LZ), CD68 and lactoferrin (LF), represent classical immunohistology marker molecules. Additionally, flow cytometry can be used to detect and quantify their expression at the single cell level in phenotypically defined leukocyte subsets. Recent results demonstrated that expression densities of these intracellular proteins vary among myeloid cell subsets, thus enabling insights into novel subset biology and development. Additionally, whole blood staining protocols allow detection of lysosomal proteins in infrequent leukocyte subsets such as circulating CD34+ hematopoietic progenitors and dendritic cells (DC). Thus, information on leukocyte subset distribution and aberrant phenotypes might be gained for diagnositic purposes. Finally, FACS detection of MPO and LZ proved to be of high value for the lineage diagnosis of acute leukemias. PMID- 15786702 TI - The use of CD38 expression by monoclonal B lymphocytes as a prognostic factor in B-cell chronic lymphocytic leukemia. AB - In B-cell chronic lymphocytic leukemia (B-CLL) the Rai and Binet staging criteria are not always able to accurately predict the prognosis of each patient. Rapidly evolving, violent disease is often seen in the so-called "good-prognosis" group, which highlights the need of additional and more refined prognostic markers. Several of these markers are described in the literature, with varying abilities to predict patient survival. Among the promising prognostic markers is flowcytometric analysis of CD38 on the monoclonal B cells in CLL. Several studies have shown that expression of CD38 is associated with a decreased overall-, or progression free survival. CD38 expression may be analyzed as percentage positive cells or as antibodies bound per cell. Addition of CD38 to the flow cytometry antibody panel for B-CLL analysis is a relatively easy way to obtain important prognostic information. PMID- 15786703 TI - Current challenges in human embryonic stem cell research: directed differentiation and transplantation tolerance. AB - Research towards potential curative transplantation of human embryonic stem (hES) cell-derived grafts in a variety of diseases has become an important topic since the successful derivation and propagation of hES cells from the inner cell mass of a blastocyst. However, clinical applicability can only be established after intensive laboratory studies that should elaborate on two major topics: A) the development of efficient, controlled and stable hES cell differentiation protocols for any specific cell type, and B) the induction of immunological tolerance against transplanted allogeneic hES cell-derived cell types. This review will briefly discuss: A) current possibilities in hES cell differentiation, followed by the development of viral, DNA and mRNA-based gene transfer strategies for hES cells, and B) possible immune modulation strategies for inducing immune tolerance against allogeneic hES cell transplants. PMID- 15786704 TI - Molecular cytometry. AB - The first flow cytometric analyses were of total cellular DNA or protein content. The advent of labeled antibodies, first polyclonal and then monoclonal, led to the analysis of cellular content of specific antigen molecules and a myriad of immunological applications. Although, strictly speaking, it is incorrect to use the word "cytometry" for anything that does not refer to cellular measurements, the term has been used for a variety of non-cellular measurements such as chromosome analysis and solution molecular measurements. In recent years, there has been an increase in the development of methods to analyze molecules and their properties in solution. These analyses have ranged from ensemble measurements, primarily fluorescence-based immunoassays, to single molecule detection with applications in DNA analysis. PMID- 15786705 TI - Molecular aspects of individual radiosensitivity. AB - Radiation therapy is a clinical treatment modality where ionizing radiation is used to treat patients with malignant neoplasms. The goal is to deliver a measured dose of radiation to a defined volume with minimal damage to surrounding normal tissue, resulting in eradication of the tumor. Radiotherapy is generally given in divided doses or fractionated. Molecular biology methods have enhanced our ability to investigate the response of cells to ionizing radiation. These methods can be applied to tissue-culture systems or to biopsies from patients both to develop a quick and easy way to predict the radiosensitivity of a patient and to understand how cells respond to stress produced by ionizing radiation. In this review we will mainly explain two major mechanisms involved in human individual radiosensitivity: the DNA-damage repair defect mechanism and the DNA repair signaling via cell cycle checkpoint defect. PMID- 15786706 TI - Gene expression in response to ionizing radiation: an overview of molecular features in hematopoietic cells. AB - We review the molecular mechanisms involved in response to ionizing radiation in various hematopoietic cell types. First, a general overview of the radiation induced cell signaling molecules in mammals is given. The importance of highly conserved kinases, such as ATM and ATR, as well as the p53 protein for maintaining the genome stability is highlighted. Next, particular attention is given to radiation-induced gene expression in the hematopoietic system. For some hemopoietic cell subpopulations, recent data are provided which might explain the differential radiosensitivity of these cells. Finally, radiation-induced cytokines are reviewed, as they affect the global response to radiation. PMID- 15786707 TI - Semiquantitative cytochemistry in evaluation of apoptotic capacity in bronchoalveolar lavage of smokers and patients with non-small-cell lung cancer. AB - Apoptotic capacity of pulmonary tissue to produce or remove apoptotic cells by alveolar macrophages (ALMs) was investigated in three groups: healthy volunteers, smokers and patients with non-small-cell lung cancer (NSCLC). Differential cell counting of bronchoalveolar lavage (BAL) specimens revealed significantly higher percentages of neutrophils and eosinophils and decreased percentage of macrophages in BAL of patients with NSCLC in comparison with nonsmokers and smokers. Proportion of lymphocytes was significantly higher in patients with NSCLC than in smokers. These changes in the BAL cell profile may reflect immunology of the lung in pulmonary malignancies. BAL eosinophils were significantly lower and AMs increased in smokers in comparison with nonsmokers. This result might be understood as a consequence of changed tissue architecture of pulmonary tissue in situ, influenced by smoking. Apoptotic detection in cytocentrifuge preparations of BAL cell suspensions was evaluated by TUNEL method. Subsequent steps, adsorption, internalization and digestion of apoptotic cells by alveolar macrophages (AMs) were estimated by semiquantitative cytochemical scoring and indexing method and correlated with percent of free apoptotic cells. Significant increase of apoptotic capacity of pulmonary tissue in control smokers (289.55+/-50.77) in comparison with that of non-smokers (218.29+/-56.24) could be a consequence of stimulated digestion inside the AMs; decreased apoptotic capacity of pulmonary tissue in NSCLC (150.30+/-40.61; p<0.05), in comparison with non-smokers and smokers is in relation to a reduced phagocytosis of the apoptotic remnants, which might be either the cause or the consequence of the oncogenic process. PMID- 15786708 TI - Inositide-modifying enzymes: a cooperative role in regulating nuclear morphology during differentiation of myeloid cells. AB - Differentiation and functional response of mature myeloid cells require cytoskeleton remodelling in a dynamic system that involves subcellular organization and regional signalling. Within the myeloid lineage, neutrophils constitute a cell type in which different cell compartments, and predominantly the nucleus, undergo distinctive large changes involving actin reorganization. In the context of the progressive elucidation of the nuclear structure and composition that has been achieved in the last two decades, it is now clear that the nucleus possesses an ordered and dynamic skeletal structure which shares many properties with the cytoskeleton, and the full set of substrates and enzymes that participate in the inositol lipid metabolism. Consolidated evidence indicate that the changes in cytoskeleton assembly are regulated also by phosphoinositides in a way dependent on their local concentration and availability. Indeed, enzymes able to affect the amount and phosphorylation of inositol lipids can play fundamental roles in determining the architectural transitions of the cell. The expression pattern and the changes of activity of PLC and PI 3-K in the nucleus during differentiation of tumoral myeloid precursors suggest that these enzymes play a crucial role in modifying the intranuclear pool of phosphoinositides, which in turn induce the changes in nucleoskeleton associated to granulocytic maturation. It can be speculated that defective control of nucleoskeleton assembly is one of the causes of dysregulated cell maturation or differentiative block in the course of myeloid leukemias. Inositide modifying enzymes can thus be regarded as potential targets for molecularly designed therapeutic intervention on hematological malignancies. PMID- 15786709 TI - Biphenotypic acute leukemia: a case report. AB - We describe an uncommon case of acute leukemia in which leukemic blasts expressed myeloid antigens and cyCD79alpha molecule. In this 49-year old male patient, two distinct blast populations were detected in peripheral blood and bone marrow samples: one of small size resembling lymphoblasts and another with pink cytoplasmic granules resembling myeloblasts. Cytochemical reaction for myeloperoxidase was negative in both cell types. Conventional cytogenetic analysis showed a normal karyotype (46 XY) in all metaphases studied, while gene rearrangement analysis by seminested PCR of the immunoglobulin heavy chain (Ig-H) and T-cell-gamma chain (TCR-gamma) receptor, showed a germline configuration of the TCR and clonal rearrangement of Ig-H chain genes. Multicolour cytofluorimetric analysis showed that bone marrow and peripheral blood blasts expressed CD19, CD79alpha bright, CD22 and terminal deoxynucleotidyl transferase (TdT) as lymphoid markers, CD13, CD117, CD15 as myeloid markers, CD34, HLA-DR as stem cell markers. CD33 myeloid antigen was expressed by 50% of the blastic population. No differences in the immunophenotypic profile were detected in the two blast populations which were identified by morphology. According to EGIL (European Group of Immunological Classification of Leukemias) and WHO (World Health Organization) criteria, a diagnosis of biphenotypic acute leukemia (BAL) was made. The patient was treated with AML induction therapy followed by autologous stem cell transplantation, but relapse free survival was 6 months. The patient died a few weeks later due to unresponsiveness to salvage chemotherapy regimens. We conclude that patients with BAL should have a risk stratification with treatment tailored to their immunophenotype and gene rearrangement profiles. PMID- 15786710 TI - Immunophenotypic approach to the identification and characterization of clonal plasma cells from patients with monoclonal gammopathies. PMID- 15786711 TI - Chronic L-arginine supplementation improves endothelial cell vasoactive functions in hypercholesterolemic and atherosclerotic monkeys. AB - Chronic exposure to L-arginine results in regression of atherosclerotic lesions and reversal of endothelial dysfunction. We investigated whether chronic L arginine supplementation induces regression of atherosclerotic lesions and reversal of endothelial dysfunction in atherogenic rhesus monkeys and the mechanism which leads to these effects. About 12 male rhesus monkeys were fed 1% cholesterol and 18 g butter for 6 months to create an experimental model of hypercholesterolaemia and atherosclerosis (Group I) and 12 monkeys were fed standard stock diet for 6 months (Group II). After, 6 months these two groups were further divided into 2 sub-groups which in addition to their respective diets were fed 2.5% L-arginine in drinking water for additional 6 months (Group III and Group IV). Systemic nitric oxide (NO) formation was assessed as plasma nitrite and cGMP formation every 3 months. Oxygen free radical (OFR) generation and malondialdehyde production as an index of lipid peroxidation were determined. Changes in isometric tension were compared in isolated ring segments of thoracic aorta from normal and hypercholesterolemic animals. Cholesterol feeding progressively reduced plasma nitrite and cGMP generation (p < 0.05). Dietary L arginine partly restored the levels of plasma nitrite and cGMP (p < 0.05) but did not change plasma cholesterol levels. L-arginine significantly reduced aortic intimal thickening, blocked the production of carotid and coronary intimal plaques and completely preserved endothelium-dependent vasodilator function. Further, L-arginine significantly inhibited generation of the reactive oxygen species (ROS) generation and lipid peroxidation. Chronic oral supplementation with L-arginine blocks the progression of plaques via restoration of nitric oxide synthase substrate availability and reduction of vascular oxidative stress. PMID- 15786712 TI - Leupaxin binds to PEST domain tyrosine phosphatase PEP. AB - PEST domain tyrosine phosphatase (PEP) is an intracellular protein tyrosine phosphatase and characterized by PEST motifs and proline-rich domains in the carboxyl terminal half. PEP is primarily expressed in hematopoietic cells, and together with PEP-binding Csk, may act as a negative regulator of antigen receptor signaling in lymphocytes. Here, we show the binding capability of PEP for leupaxin, which is preferentially expressed in hematopoietic cells and a comparatively new member of the paxillin family characterized by two protein protein interaction modules, LIM domains and LD motifs. These results suggested that leupaxin might participate in the regulation of the signaling cascade through the binding to PEP in lymphocytes. PMID- 15786713 TI - Myocardial protective effect of octylguanidine against the damage induced by ischemia reperfusion in rat heart. AB - This study shows that the hydrophobic cation octylguanidine protects against myocardial damage induced by ischemia-reperfusion. The protective effect of the amine was analyzed after 5 min of coronary occlusion followed by 5 min reperfusion in rat hearts. ECG tracings from rats treated with an i.v., injection of 5 mg/kg of octylguanidine showed a total absence of post-reperfusion arrhythmias, conversely to what was observed in untreated rats. The histological images showed that myocardium fibers from treated rats were in good shape and retained their striae, also there was absence of edema. Furthermore, the accumulation of 201Tl in hearts from these rats indicated that the tissue did not suffer disruption or impairment in membrane functions. The above correlated with the fact that mitochondria isolated from the ventricular free wall from treated rats preserved their ability to synthesize ATP. We propose that the protective effect of octylguanidine might be due to its documented inhibitory action on the opening of mitochondrial non-specific pores, a mechanism which is associated in heart injury as induced by reperfusion. PMID- 15786714 TI - A thermostable beta-ketothiolase of polyhydroxyalkanoates (PHAs) in Thermus thermophilus: purification and biochemical properties. AB - Polyhydroxyalkanoates (PHAs) are polyesters of hydroxyalkanoates (HAs) synthesised by numerous bacteria as intracellular carbon and energy storage compounds which accumulate as granules in the cytoplasm of the cells. The biosynthesis of PHAs, in the thermophilic bacterium T. thermophilus grown in a mineral medium supplemented with sodium gluconate as sole carbon source has been recently reported. Here, we report the purification at apparent homogeneity of a beta-ketoacyl-CoA thiolase from T. thermophilus, the first enzyme of the most common biosynthetic pathway for PHAs. B-Ketoacyl-CoA thiolase appeared as a single band of 45.5-kDa molecular mass on SDS/PAGE. The enzyme was purified 390 fold with 7% recovery. The native enzyme is a multimeric protein of a molecular mass of approximately of 182 kDa consisting of four identical subunits of 45.5 kDa, as identified by an in situ renaturation experiment on SDS-PAGE. The enzyme exhibited an optimal pH of approximately 8.0 and highest activity at 65 degrees C for both direction of the reaction. The thiolysis reaction showed a substrate inhibition at high concentrations; when one of the substrates (acetoacetyl CoA or CoA) is varied, while the concentrations of the second substrates (CoA or acetoacetyl CoA respectively) remain constant. The initial velocity kinetics showed a pattern of a family of parallel lines, which is in accordance with a ping-pong mechanism. beta-Ketothiolase had a relative low Km of 0.25 mM for acetyl-CoA and 11 microM and 25 microM for CoA and acetoacetyl-CoA, respectively. The enzyme was inhibited by treatment with 1 mM N-ethylmaleimide either in the presence or in the absence of 0.5 mM of acetyl-CoA suggesting that possibly a cysteine is located at/or near the active site of beta-ketothiolase. PMID- 15786715 TI - Homocysteine inhibits store-mediated calcium entry in human endothelial cells: evidence for involvement of membrane potential and actin cytoskeleton. AB - The role of homocysteine for store-operated calcium influx was investigated in human umbilical cord endothelial cell line. Homocysteine significantly decreased thapsigargin-evoked Ca2+ entry, membrane hyperpolarization and actin polymerization. GSH and DTT prevented homocysteine-induced inhibition of thapsigargin-evoked Ca2+ entry, membrane hyperpolarization and actin polymerization; while GSSG had the opposite effect. Homocysteine blocked large conductance Ca2+-activated K+ (BK(Ca)) channels in a concentration-dependent manner and related to the redox status of the endothelial cells. BK(Ca) channels opener NS1619 reversed thapsigargin-evoked Ca2+ entry, membrane hyperpolarization and actin polymerization; BK(Ca) channels inhibitor iberiotoxin had the opposite effect. The findings suggest that homocysteine is involved in store-regulated Ca2+ entry through membrane potential-dependent and actin cytoskeleton-dependent mechanisms, redox status of homocysteine and BK(Ca) channels may play a regulatory role in it. PMID- 15786716 TI - Reduced inhibitor 1 and 2 activity is associated with increased protein phosphatase type 1 activity in left ventricular myocardium of one-kidney, one clip hypertensive rats. AB - In failing hearts, although protein phosphatase type 1 (PP1) activity has increased, information about the regulation and status of PP1 inhibitor-1 (INH-1) and inhibitor-2 (INH-2) is limited. In this study, we examined activity and protein expression of PP1, INH-1 and INH-2 and phosphorylation of sarcoplasmic reticulum (SR) phospholamban (PLB), a substrate of PP1 and modulator of SR Ca2+ ATPase activity, in failing and non-failing hearts. These studies were performed in LV myocardium of seven rats with chronic renal hypertension produced by Goldblatt's one-kidney, one-clip procedure and seven age-matched sham-operated normal controls (CTR). Eight weeks after surgery, LV ejection fraction, LV hypertrophy, and pulmonary congestion were determined in all rats. PP1 activity (nmol 32P/min/mg non-collagen protein) was assessed in LV homogenates using 32P labeled phosphorylase a as substrate. INH-1 and INH-2 activity was determined in the immunoprecipitate of LV homogenates and expressed as percentage inhibitory activity. Using a specific antibody, LV tissue levels of PP1C and calsequestrin (CSQ), a SR calcium binding protein, which is not altered in failing hearts, were also determined. Further, total and phosphorylated PLB, INH-1 and INH-2 protein levels were determined in the LV homogenate and phosphoprotein-enriched fraction, respectively. The band density of each protein was quantified in densitometric units and normalized to CSQ. RESULTS: rats with chronic renal hypertension exhibited significantly reduced LV ejection fraction and increased LV hypertrophy and pulmonary congestion, characteristics of chronic heart failure (CHF). We found that compared to CTR, (1) both INH-1 (10.2+/-2 versus 57.5+/-1; p < 0.05) and INH-2 activity (3.8+/-0.4 versus 36.2+/-4; p < 0.05) were reduced, (2) total and phosphorylated PLB amount reduced, (3) protein level of phosphorylated INH-1 was reduced (2.32+/-0.1 versus 0.73+/-0.04; p < 0.05) whereas that of phosphorylated INH-2 increased (3.05+/-0.3 versus 1.42+/-0.1; p < 0.05), and (4) PP1 activity was increased approximately 2.6-fold in rats with CHF (1.59+/-0.05 versus 0.61+/-0.01; p < 0.05) while protein level of the catalytic subunit of PP1 (PP1C) increased 3.85-fold (0.77+/-0.05 versus 0.20+/-0.02; p < 0.05). These results suggest that reduced inhibitory INH-1 and INH-2 activity, increased PP1C protein level, and reduced PLB phosphorylation are associated with increased PP1 activity in failing hearts. PMID- 15786717 TI - Contribution of the PI 3-kinase/Akt survival pathway toward osmotic preconditioning. AB - Osmolytes are rapidly lost from the ischemic heart, an effect thought to benefit the heart by reducing the osmotic load. However, the observation that chronic lowering of one of the prominent osmolytes, taurine, is more beneficial to the ischemic heart than acute taurine loss suggests that osmotic stress may benefit the ischemic heart through multiple mechanisms. The present study examines the possibility that chronic osmotic stress preconditions the heart in part by stimulating a cardioprotective, osmotic-linked signaling pathway. Hyperosmotic stress was produced by treating rat neonatal cardiomyocytes during the pre hypoxic period with either the taurine depleting agent, beta-alanine (5 mM), or with medium containing 25 mM mannitol. The cells were then subjected to chemical hypoxia in medium containing 3 mM Amytal and 10 mM deoxyglucose but lacking beta alanine and mannitol. Cells that had been pretreated with either 5 mM beta alanine or 25 mM mannitol exhibited resistance against hypoxia-induced apoptosis and necrosis. Associated with the osmotically preconditioned state was the activation of Akt and the inactivation of the pro-apoptotic factor, Bad, both events blocked by the inhibition of PI 3-kinase. However, preconditioning the cardiomyocyte with mannitol had no effect on the generation of free radicals during the hypoxic period. Osmotic stress also promoted the upregulation of the anti-apoptotic factor, Bcl-2. Since inhibition of PI 3-kinase with Wortmannin also prevents osmotic-mediated cardioprotection, we conclude that hyperosmotic mediated activation of the PI 3-kinase/Akt pathway contributes to osmotic preconditioning. PMID- 15786718 TI - Chlamydia pneumoniae infected macrophages exhibit enhanced plasma membrane fluidity and show increased adherence to endothelial cells. AB - Chlamydia pneumoniae, an intracellular prokaryote, is known to have requirement for some lipids which it is incapable of synthesizing, and these lipids have important fluidizing roles in plasma membrane. We decided to examine if the trafficking of these lipids to C. pneumoniae alters the physicochemical properties of macrophage plasma membrane, affects the expression of genes and proteins of enzymes associated with metabolism of some of these lipids and assess if Ca2+ signaling usually induced in macrophages infected with C. pneumoniae modulates the genes of these selected enzymes. Chlamydia pneumoniae induced the depletion of macrophage membrane cholesterol, phosphatidylinositol and cardiolipin but caused an increase in phosphotidylcholine resulting in a relative increase in total phospholipids. There was increased membrane fluidity, enhanced macrophage fragility and heightened adherence of macrophages to endothelial cells despite the application of inhibitor of adhesion molecules. Also, there was impairment of macrophage 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase gene and protein expression independent of Ca2+ signaling, while phospholipase C gene and protein were up-regulated in a manner minimally dependent on Ca2+ signaling. The implications of these findings are that macrophages infected with C. pneumoniae have altered membrane physicochemical characteristics which may render them atherogenic. PMID- 15786719 TI - Molecular cloning and characterization of a novel muscle adenylosuccinate synthetase, AdSSL1, from human bone marrow stromal cells. AB - Vertebrates have muscle and non-muscle isozymes of adenylosuccinate synthetase (AdSS, EC 6.3.4.4), which catalyzes the first committed step in AMP synthesis. A novel muscle isozyme of adenylosuccinate synthetase, human AdSSL1, is identified from human bone marrow stromal cells. AdSSL1 is 98% identical to mouse muscle type AdSS1 and contains conserved sequence and structural features of adenylosuccinate synthetase. Human AdSSL1 gene is mapped to chromosome 14p32.33. After stimulation, leukemia cells express AdSSL1 in a time-dependent manner different from that of non-muscle adenylosuccinate synthetase. The human AdSSL1 is predominantly expressed in skeletal muscle and cardiac tissue consistent with the potential role for the enzyme in muscle metabolism. Overexpressed AdSSL1 protein in COS-7 cells locates in cytoplasm. Recombinant AdSSL1 protein possesses typical enzymatic activity to catalyze adenylosuccinate formation. The identification of human AdSSL1 with predominant expression in muscle tissue will facilitate future genetic and biochemical analysis of the enzyme in muscle physiology. PMID- 15786720 TI - Increased expression of IL-6 and LIF in the hypertrophied left ventricle of TGR(mRen2)27 and SHR rats. AB - Cytokines from the interleukin-6 (IL-6) family have been reported to play an important synergistic role with angiotensin II in the development of pathological cardiac hypertrophy. Whether their expression pattern changes in vivo, in an angiotensin I-dependent hypertrophied myocardium has not been reported. In this study, we addressed that issue using two animal models of angiotensin II dependent cardiac hypertrophy. Heterozygous transgenic TGR(mRen2)27 (TGR) with an overactive cardiac renin angiotensin system and the closely related spontaneously hypertensive rats (SHR) were compared to their respective control rats. The mRNA levels of IL-6, leukemia inhibitory factor (LIF), ciliary neurotrophic factor (CNTF) and cardiotrophin-1 (CT-1) as well as their receptor subunits, glycoprotein 130 (gp130), IL-6 receptor (IL-6R), LIFR, and CNTFR, were measured by semi-quantitative RT-PCR. The protein levels of IL-6, LIF and CT-1 were investigated by western blot. TGR and SHR both displayed significant over expression of mRNA and protein levels for IL-6 and LIF. In TGR, the increased level of LIF was accompanied by a decrease in mRNA levels for LIFR and CNTFR. In SHR, a higher level of mRNA IL-6R was observed. By contrast, the mRNA and protein levels for CT-1 and the mRNA level for gp130 did not vary in these two models. These findings suggest that IL-6 and LIF, but not CT-1, contribute to angiotensin II-dependent left ventricular hypertrophy in the two hypertensive rat models, TGR(mRen2)27 and SHR. PMID- 15786721 TI - Effects of single high-dose and multiple low-dose streptozotocin on contraction and intracellular Ca2+ in ventricular myocytes from diabetes resistant and susceptible rats. AB - Administration of a single high-dose (SHD) of streptozotocin (STZ) to young adult rats causes a diabetic cardiomyopathy. Albino Oxford (AO) and Dark Agouti (DA) inbred strains of rats are susceptible to developing diabetes when administered a SHD of STZ but differ in susceptibility to multiple low-dose (MLD) STZ. We have investigated the effects of SHD and MLD-STZ on contraction and intracellular Ca2+, measured with fura-2, in ventricular myocytes from AO and DA rats at 18-20 weeks after treatment. Time to peak shortening was significantly prolonged in myocytes from DA rats after SHD-STZ but was not altered in DA rats after MLD-STZ or in AO rats by either MLD or SHZ-STZ treatment. Time to peak shortening in myocytes from DA control and DA rats after SHD-STZ were 88+/-2 ms and 107+/-4 ms, respectively. Time to half relaxation and the amplitude of myocyte shortening were not altered in AO or DA rats by either MLD or SHD-STZ treatment. Amplitude, time to peak fura-2 transient and time to half relaxation of the fura-2 transient were not significantly altered in AO or DA rats by either MLD or SHD-STZ treatment. Contractile defects reported in myocytes from SHD-STZ treated DA rats may be a consequence of altered myofilament sensitivity to Ca2+. The hyperglycaemic effects of MLD-STZ and SHD-STZ induced diabetes was much greater in DA compared to AO rats and the effects of the hyperglycaemia on the time course of ventricular myocyte contraction was most profound in DA rats after SHD STZ. PMID- 15786722 TI - Mevastatin induces apoptosis in HL60 cells dependently on decrease in phosphorylated ERK. AB - Mevastatin which is an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG CoA) reductase, the rate-limiting enzyme in cholesterol synthesis, suppress cell proliferation and induce apoptosis. However, the molecular mechanism of apoptosis induction is not well understood. So, in the present study, we attempted to clarify the mechanism by which mevastatin induces apoptosis in HL60 cells. It was found that mevastatin induced apoptosis. At that time, we observed an increase in caspase-3 activity and morphological fragmentation of the nuclei. The apoptosis induced by mevastatin was not inhibited by the addition of farnesyl pyrophosphate (FPP), squalene, ubiquinone, and isopentenyladenine, but was inhibited by the addition of geranylgeranyl pyrophosphate (GGPP). When we examined the survival signals at the time of apoptotic induction, we also observed that the administration of mevastatin had caused a remarkable decrease in the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). However, other survival signals, such as nuclear factor kappa B (NF-kappaB), protein kinase B (Akt), and p38 mitogen-activated protein kinase (p38), exhibited no change. In addition, no quantitative change was observed in Bcl-2, which was an anti-apoptosis protein. It was also observed that apoptosis was induced when U0126, an MEK inhibitor, was added to the cells to inhibit ERK. These results suggested that mevastatin induced apoptosis when it inhibited GGPP biosynthesis and consequently decreased the level of phosphorylated ERK, which was a survival signal; moreover, at that time, there was no influence on NF-kappaB, Akt, p38, and Bcl-2. The results of this study also suggested that mevastatin could be used as an anticancer agent. PMID- 15786723 TI - Prevention of cataract by pyruvate in experimentally diabetic mice. AB - Previous studies have demonstrated that administration of pyruvate prevents cataract formation in diabetic rats. It is known that the induction of cataractous process in this case is initiated by aldose reductase (AR) catalyzed synthesis and accumulation of excessive sorbitol in the lens fibres and epithelium and their consequent osmotic hydration. Synthesis of this and other polyols is competitively inhibited by pyruvate. The objective of the present investigations was hence to determine whether pyruvate would have a similar protective effect in species where cataract formation is relatively independent of sorbitol synthesis such as in humans where the lens AR activity is extremely low, especially with glucose as a substrate. The Km of AR for glucose is known to be very high. The possible protective effect of pyruvate in the low AR models was conceived on the basis of our previous findings suggesting that it can also exert substantial antiglycating as well as antioxidant effects. The present studies have hence been conducted with mice, a species known to be low in lens AR, similar to that in humans. As stipulated, pyruvate administration has indeed been found to offer a significant protection against development of diabetic cataract in this model also. The effect correlated with the inhibition of protein glycation as well as of oxidative stress. The latter was apparent by the prevention of the loss of glutathione known to be associated with diabetes. Although there was a small but noticeable increment in the sorbitol content of the diabetic lenses, this was osmotically insignificant. Even this increase was prevented by pyruvate. The magnitude of the elevation in the contents of glycated proteins and the depression in the level of glutathione were, on the contrary, highly pronounced, suggesting a more prominent role of the latter factors. In addition, the possibility of a direct metabolic support it could offer to the tissue is also imminent by its effect on the maintenance of ATP, as shown earlier. The present studies are therefore considered more relevant to the pathogenesis of cataract in human diabetics and its possible prevention by endogenous compounds with antiglycating and antioxidant properties. Inhibition of cataract formation by pyruvate in an animal model with low lens AR, similar to that in humans, has been shown for the first time. PMID- 15786724 TI - Interpretation of relevance of sodium-calcium exchange in action potential of diabetic rat heart by mathematical model. AB - Sarcolemmal Na+-Ca2+ exchange plays a central role in ion transport of the myocardium and the current carried with it contributes to the late phase of the action potential (AP) besides the contribution of outward K+-currents. In this study, the mathematical model for AP of the diabetic rat ventricular myocytes [34] was modified and used for the diabetic rat papillary muscle. We used our experimentally measured values of two K+-currents; transient outward current, Ito and steady-state outward current, Iss, as well as L-type Ca2+-current, I(CaL), then compared with the simulated values. We have demonstrated that the prolongation in the AP of the papillary muscle of the diabetic rats are not due to the alteration of I(CaL) but mainly due to the inhibition of the K+-currents and also the Na+-Ca2+ exchanger current, I(Na-Ca). In combination with our experimental data on sodium-selenite-treated diabetic rats, our simulation results provide new information concerning plausible ionic mechanisms, and second a possible positive effect of selenium treatment on the altered I(Na-Ca) for the observed changes in the AP duration of streptozotocin-induced diabetic rat heart. PMID- 15786726 TI - Effects of angiotensin-II receptor blocker candesartan cilexetil in rats with dilated cardiomyopathy. AB - We examined effects of an angiotensin-II receptor blockers, candesartan cilexetil, in rats with dilated cardiomyopathy after autoimmune myocarditis. Candesartan cilexetil showed angiotensin-II blocking action in a dose-dependent manner in rats with dilated cardiomyopathy. Twenty-eight days after immunization, surviving Lewis rats were divided into four groups and given candesartan cilexetil at 0.05 mg/kg, 0.5 mg/kg or 5 mg/kg per day (Group-C0.05, n = 15, Group C0.5, n = 15 and Group-C5, n = 15, respectively) or vehicle alone (Group-V, n = 15). After oral administration for 1 month, the left ventricular end-diastolic pressure and heart weight/body weight ratio were lower in Group-C0.05 (13.3+/-1.1 mmHg and 3.7+/-0.2 g/kg, respectively), in Group-C0.5 (8.0+/-0.9 mmHg and 3.3+/ 0.1 g/kg, respectively) and in Group-C5 (5.5+/-1 mmHg and 3.1+/-0.1 g/kg, respectively) than in Group-V (13.5+/-1.0 mmHg and 3.8+/-0.2 g/kg, respectively). The area of myocardial fibrosis was also lower in Group-C0.05 (25+/-3%), in Group C0.5 (20+/-3%), and in Group-C5 (12+/-1%) than in Group-V (32+/-4%). Furthermore, expressions of transforming growth factor-beta1 and collagen-III mRNA were suppressed in Group-C0.05 (349+/-23% and 395+/-22%, respectively), Group-C0.5 (292+/-81% and 364+/-42%, respectively) and in Group-C5 (204+/-63% and 259+/-33%, respectively) compared with those in Group-V (367+/-26% and 437+/-18%, respectively). These results suggest that candesartan cilexetil can improve the function of inefficient heart. PMID- 15786725 TI - Effect of Tridax procumbens on liver antioxidant defense system during lipopolysaccharide-induced hepatitis in D-galactosamine sensitised rats. AB - The present study was carried out to assess the effect of chloroform insoluble fraction of ethanolic extract of Tridax procumbens (TP) against D Galactosamine/Lipopolysaccharide (D-GalN/LPS)-induced hepatitis in rats. Induction of rats with D-GalN/LPS (300 mg/kg body weight/30 microg/kg body weight) led to a marked increase in lipid peroxidation as measured by thiobarbituric acid reactive substances (TBARS) in liver. Further there was a decline in the activities of enzymic antioxidants such as superoxide dismutase, catalase, glutathione peroxidase, glutathione s-transferase and the levels of non enzymic antioxidants namely reduced glutathione, vitamin C and vitamin E. These biochemical alterations were normalised upon pretreatment with TP extract. Thus, the above results suggest that TP (300 mg/kg body weight orally for 10 days) is very effective in allievating the D-GalN/LPS-induced oxidative stress suggesting its antioxidant property. PMID- 15786727 TI - Ca2+-induced permeability transition in human lymphoblastoid cell mitochondria from normal and Huntington's disease individuals. AB - Huntington's disease (HD) is associated with expansion of polyglutamine tract in a protein named huntingtin (htt) that is expressed in virtually all body tissues. Thus mutated htt (HD-htt) might affect all organs, although clinical manifestations of HD are associated with selective loss of corticostriatal neurons of the brain. In this work we studied how HD-htt affects mitochondria in human peripheral blood cells. We compared various functions of mitochondria isolated from cultured lymphoblastoid cells derived from three HD patients with juvenile onset of the disease (HD-LBM) and three age-matched control (C-LBM) individuals. Respiratory parameters in different metabolic states, with succinate and glutamate plus malate were the same for all control and HD cell lines. State 4 membrane potential in HD-LBM was slightly lower than in C-LBM. The calcium retention capacity (CRC) of mitochondria was estimated using simultaneously several methods to register permeability transition (PT). We found that LBM do not undergo swelling upon Ca2+-induced PT, and do not increase CRC in the presence of ADP + oligomycin. Although each cell line had different CRC values, qualitatively PT was different in C-LBM and HD-LBM. With C-LBM cyclosporin A (CsA) increased CRC significantly, while with HD-LBM CsA was ineffective. In C LBM depolarization of mitochondria and a large pore opening (PT) always occurred simultaneously. In HD-LBM depolarization occurred at 20-50% lower Ca2+ loads than PT. We suggest that HD-htt promotes low H+ conductance of the mitochondria by interacting with proteins at the contacts sites without directly promoting PT or hampering mitochondrial oxidative phosphorylation. PMID- 15786728 TI - The L-type calcium channel alpha 1C subunit gene undergoes extensive, uncoordinated alternative splicing. AB - The alpha1C subunit is the pore-forming protein for the L-type calcium channel. Previous studies indicate that there is possible tissue-specific alternative splicing of this gene. In this study we cloned the entire open reading frame of the alpha1C subunit cDNA from adult rat cardiac myocytes in a single piece (6.64 kb). Using 75 positive clones that were identified by restriction enzyme mapping, we tested the alternative splicing patterns of the Ca(v) 1.2 gene that encodes the alpha1C subunit protein and focused on five loci: IS6, post-IS6, IIIS2, IVS3, and the c-terminus. The results indicate that: (1) alternative splicing occurs in most of the loci, giving rise to two or three different isoforms at those sites; (2) there is a predominant form for each splicing site, (3) there does not appear to be consistent coordination of splicing at multiple loci of this gene. Alternative splicing is not tissue-specific in most regions. PMID- 15786729 TI - H2O2-induced changes in mitochondrial activity in isolated mouse pancreatic acinar cells. AB - This study employed confocal laser scanning microscopy to monitor the effect of H2O2 on cytosolic as well as mitochondrial calcium (Ca2+) concentrations, mitochondrial inner membrane potential (psi m) and flavine adenine dinucleotide (FAD) oxidation state in isolated mouse pancreatic acinar cells. The results show that incubation of pancreatic acinar cells with H2O2, in the absence of extracellular Ca2+ ([Ca2+],) led to an increase either in cytosolic and in mitochondrial Ca2+ concentration. Additionally, H2O2 induced a depolarization of mitochondria and increased oxidized FAD level. Pretreatment of cells with the mitochondrial inhibitors rotenone or cyanide inhibited the response induced by H2O2 on mitochondrial inner membrane potential but failed to block oxidation of FAD in the presence of H2O2. However, the H2O2-evoked effect on FAD state was blocked by pretreatment of cells with the mitochondrial uncoupler, carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP). On the other hand, perfusion of cells with thapsigargin (Tps), an inhibitor of the SERCA pump, led to an increase in mitochondrial Ca2+ concentration and in oxidized FAD level, and depolarized mitochondria. Pretreatment of cells with thapsigargin inhibited H2O2 evoked changes in mitochondrial Ca2+ concentration but not those in membrane potential and FAD state. The present results have indicated that H2O2 can evoke marked changes in mitochondrial activity that might be due to the oxidant nature of H2O2. This in turn could represent the mechanism of action of ROS to induce cellular damage leading to cell dysfunction and generation of pathologies in the pancreas. PMID- 15786730 TI - Cloning and expression of PPAR-gamma and PGC-1alpha from the hibernating ground squirrel, Spermophilus tridecemlineatus. AB - The peroxisome proliferator-activated receptor (PPAR) family of transcription factors play a key role in lipid metabolism and have been implicated in a number of disease states, most notably of which is obesity. Controlled regulation of lipid metabolism is a key ingredient for successful hibernation. Partial cDNA sequences for one of the PPAR proteins, PPARgamma and the PPARgamma co-activator (PGC-1alpha) have been cloned from the hibernating ground squirrel, Spermophilus tridecemlineatus and show differential regulation during hibernation at the mRNA level using relative RT-PCR and at the protein level via immunoblotting in brown adipose tissue (BAT), heart, skeletal muscle and white adipose tissue (WAT). The cDNA sequence for PGC-1alpha revealed a number of amino acid substitutions and two were worthy of note, one resulting in the loss of a potential protein kinase C (PKC) site, while another resulted in the creation of a PKC site, suggesting that PKC may be important in regulating PGC-1alpha. RT-PCR revealed a near 2-fold up-regulation of PPARgamma in BAT and to a lesser extent (<1.5-fold) in heart and WAT, while PGC-1alpha displayed significantly higher levels of expression in skeletal muscle during hibernation (3.1-fold, p < 0.005). The protein levels of PPARy were significantly increased in BAT and WAT (1.5 and 1.8-fold, respectively) while PGC-1alpha displayed significant changes in expression in heart (3.5-fold) and skeletal muscle (1.8-fold). Our current findings indicate a role for increased expression of PPARy and PGC-1alpha in hibernating animals. PMID- 15786731 TI - Protein phosphatase type 2A, PP2A, is involved in degradation of gp130. AB - The interleukin-6 (IL-6) stimulates growth in cells such as multiple myeloma and B-cell plasmacytomas/hybridomas, while it inhibits growth in several myeloid leukemia cells. The IL-6 receptor has subunit called gp130. It was reported that Ser-782 of gp130 is phosphorylated by unidentified kinase(s) in cell extracts, and level of gp130 (S782A) transiently expressed on the cell surface of COS-7 is 6-times higher than that of the wild type. These results motivated us to analyze whether the phosphorylation of gp130 at Ser-782 is involved in its degradation or not. In this study, we demonstrated here that treatment of HepG2 cells with okadaic acid (OA), a potent inhibitor for PP2A, promotes phosphorylation of gp 130 at Ser-782 and degradation of gp 130. MG115, a proteasome inhibitor, suppressed this degradation. These effects of OA could not be replaced with tautomycetin (TC), an inhibitor for PP1. Purified PP2A dephosphorylated phospho Ser-782 of gp130 in vitro. IL-6-induced activation of Stat3 was suppressed by preincubation of the cells with OA, suggesting that the IL-6 signaling pathway was blocked by OA through degradation of gp 130. Taken together, present results strongly suggest that degradation of gp 130 is regulated through a phosphorylation-dephosphorylation mechanism in which PP2A is crucially involved and that gp 130 is a potential therapeutic target in cancers. PMID- 15786733 TI - Opposing effects of two osmolytes--trehalose and glycerol--on thermal inactivation of rabbit muscle 6-phosphofructo-1-kinase. AB - Trehalose and glycerol are known as good stabilizers of function and structure of several macromolecules against stress conditions. We previously reported that they have comparable effectiveness on protecting two yeast cytosolic enzymes against thermal inactivation. However, enzyme protection has always been associated to a decrease in catalytic activity at the stabilizing conditions i.e., the presence of the protective molecule. In the present study we tested trehalose and glycerol on thermal protection of the mammalian cytosolic enzyme phosphofructokinase. Here we found that trehalose was able to protect phosphofructokinase against thermal inactivation as well as to promote an activation of its catalytic activity. The enzyme incubated in the presence of 1 M trehalose did not present any significant inactivation within 2 h of incubation at 50 degrees C, contrasting to control experiments where the enzyme was fully inactivated during the same period exhibiting a t0.5 for thermal inactivation of 56+/-5 min. On the other hand, enzyme incubated in the presence of 37.5% (v/v) glycerol was not protected against incubation at 50 degrees C. Indeed, when phosphofructokinase was incubated for 45 min at 50 degrees C in the presence of lower concentrations of glycerol (7.5-25%, v/v), the remaining activity was 2-4 times lower than control. These data show that the compatibility of effects previously shown for trehalose and glycerol with some yeast cytosolic enzymes can not be extended to all globular enzyme system. In the case of phosphofructokinase, we believe that its property of shifting between several different complex oligomers configurations can be influenced by the physicochemical properties of the stabilizing molecules. PMID- 15786732 TI - ARP2 a novel protein involved in apoptosis of LNCaP cells shares a high degree homology with splicing factor Prp8. AB - The mechanism of apoptosis has been recognized as an important event in processes such as cellular development and homeostasis, as well as degenerative conditions like cancer. Prostate cancer during its advanced stages develops androgen independent cells that ultimately overgrow and promote metastatic events. Our group employing androgen independent LNCaP cells have previously proposed, based on electrophysiological findings, that apoptosis induced cells overexpress a cell death calcium channel-like molecule. Here we report the cloning and expression in Xenopus laevis oocytes of apoptosis regulated protein 2 (ARP2), a protein overexpressed in apoptosis induced LNCaP cells capable to induce calcium inward currents and apoptosis typical morphology changes in oocytes injected with arp2 mRNA. Our results also indicate that clone arp2 cDNA (1.3Kb) shares a 99% homology with a small fragment that corresponds to 18% of the complete sequence of Prp8 cDNA (7.0 Kb), a molecule that codifies for an important protein in the assembly of the spliceosome. We propose that protein ARP2 as a fragment of protein Prp8, corresponds to a molecule with a new function in apoptosis related phenomena. PMID- 15786735 TI - [Role of mammalian HSP70-based chaperone system in cytosol]. PMID- 15786734 TI - Interaction of keratinocytes and fibroblasts modulates the expression of matrix metalloproteinases-2 and -9 and their inhibitors. AB - Disruption of epidermal-mesenchymal communication due to a delay in epithelialization, increases the frequency of developing fibrotic conditions in skin. As matrix metalloproteinases-2 (MMP-2) and -9 (MMP-9) are two key enzymes involved in wound healing and tissue remodeling, here we examined the efficacy of keratinocyte-fibroblast interaction on modulation of these enzymes and their inhibitors. The conditioned media derived from keratinocytes and fibroblasts grown in upper and lower chambers of a co-culture system, respectively, were analyzed for MMP-2 and -9. Keratinocyte or fibroblast conditioned medium (FCM) was used as a control. Gelatinolytic activity analyzed by zymography showed that keratinocytes mainly express MMP-9 and to a lesser extent MMP-2; while fibroblasts express only MMP-2. In a co-culture system, the activities of both MMP-2 and MMP-9 markedly increased in conditioned media collected from bottom chambers. These findings were consistent with the level of MMP-2 and MMP-9 measured by Western blot. Using the same experimental setting, the levels of tissue inhibitors of MMPs (TIMPs) secreted by keratinocytes and fibroblasts grown in the same co-culture system were also evaluated. Western blot showed that fibroblasts secrete only TIMP-1 and TIMP-2 whose levels were increased by co culturing fibroblasts with keratinocytes. In contrary the level of TIMP-3, which was mainly expressed by keratinocytes, increased by co-culturing these cells with fibroblasts. In conclusion, interaction of fibroblast-keratinocyte modulates the levels of MMP-2 and -9 and their inhibitors produced by these cells and this interaction may be critical for a better healing quality at a late stage of the wound healing process. PMID- 15786736 TI - [DNA repair in higher plants]. PMID- 15786737 TI - [Protein phosphorylation-dependent regulation of circadian timekeeping]. PMID- 15786738 TI - [A redox sensing heme-protein from Escherichia coli, Ec DOS: regulation mechanism of phosphodiesterase activity]. PMID- 15786739 TI - [The role of mDia in cytoskeletal organization]. PMID- 15786740 TI - [mDia1 and formins: processive cappers surfing along with the fast-growing actin end]. PMID- 15786741 TI - [Role of heterotrimeric G protein in asymmetric cell division of Drosophila neuroblasts]. PMID- 15786742 TI - [Disease proteomics of oxidatively induced protein carbonyls]. PMID- 15786743 TI - [Structure and function of multicopper oxidoreductases]. PMID- 15786744 TI - [Molecular mechanisms in acceptor substrate recognition of a human glucuronyltransferase, GlcAT-P, an enzyme critical in the biosynthesis of the carbohydrate epitope HNK-1]. PMID- 15786745 TI - Multidetector-row CT evaluation of aortic disease. PMID- 15786746 TI - Monitoring of respiratory movement of the diaphragm for gated radiotherapy. AB - PURPOSE: In order to obtain a respiratory signal for gated radiotherapy, diaphragm movement was monitored. MATERIALS AND METHODS: To obtain the diaphragm signal, an image brightness stabilizer for fluoroscopy of an X-ray simulator was used. The image brightness stabilizer output electric current in response to image brightness, which changed with diaphragm movement. The electric current was recorded as the diaphragm signal and compared with the abdominal wall signal, which was obtained by monitoring the height of the abdominal wall using a laser displacement sensor. RESULTS: A regular diaphragm signal was obtained in all of nine patients. At 70% expiration of the abdominal wall signal, the diaphragm signal ranged from 35% to 70% expiration (median, 55%). At 30% inspiration of the abdominal wall signal, the diaphragm signal ranged from 10% to 30% inspiration (median, 20%). CONCLUSION: A regular diaphragm signal could be conveniently obtained utilizing an image brightness stabilizer. The diaphragm signal tended to be delayed in comparison with the abdominal wall signal. PMID- 15786747 TI - Evaluation of inter- and intrafraction organ motion during intensity modulated radiation therapy (IMRT) for localized prostate cancer measured by a newly developed on-board image-guided system. AB - PURPOSE: To investigate prostatic organ motion at both setup and intrafraction using an onboard image-guided system. An intrafraction field-based repositioning method also was evaluated. MATERIALS AND METHODS: A dual fluoroscopy with amorphous-silicon flat panel (DFFP) system was used for the three-dimensional registration of implanted markers in the prostate of eight organ-confined cancer patients planned for treatment with intensity modulated radiation therapy (IMRT). Day-to-day motion errors were quantified and intrafraction displacements of more than +/-1 mm were corrected. RESULTS: Among 214 fractions and 565 system views, day-to-day mean magnitude of marker discrepancy +/- standard deviation (SD) was 1.76 +/- 1.4 mm, 3.14 +/- 1.6 mm, and 3.78 +/- 2.4 mm in the right-left, cranial caudal, and anterior-posterior directions, respectively. The intrafractional mean magnitude +/- SD of marker displacement was 0.45 +/- 0.7 mm, 1.08 +/- 1.38 mm and 1.45 +/- 1.70 mm in the right-left, cranial-caudal, and anterior-posterior directions, respectively. Intrafraction corrected sessions (84/214) showed a median (range) of motion of 0.1 mm (-1.2 to 0.7 mm), -0.2 mm (-2.1 to 1.1 mm), and -0.2 mm (-1.7 to 2.0 mm) in the right-left, cranial-caudal, and anterior posterior directions, respectively. CONCLUSION: Motion uncertainty can be considerably decreased with daily use of the DFFP system. Reduced intrafraction organ motion clearly endorsed the value of the repositioning approach, allowing a safer dose escalation protocol. PMID- 15786748 TI - Preliminary estimation of treatment effect on uterine cervical squamous cell carcinoma in terms of tumor regression rate: comparison between chemoradiotherapy and radiotherapy alone. AB - PURPOSE: We preliminarily estimated the treatment effect on cervical cancer in terms of the tumor regression rate (TRR) achieved with chemoradiotherapy and radiotherapy alone. MATERIALS AND METHODS: The study included cervical squamous cell carcinomas treated by radiotherapy alone (n=45) or chemoradiotherapy (concurrent once-a-week cisplatin 30 mg/m2, n=13). Tumors were measured three dimensionally on pre- and mid-treatment magnetic resonance images. TRR was defined as the slope of the exponential regression curve of tumor volume (day( 1)) on the assumption that tumors regressed exponentially with time. RESULTS: TRR ranged widely (0.004-0.090 day-(-1)) and did not significantly differ between treatment with chemoradiotherapy (median, 0.032 day(-1)) and with radiotherapy alone (median, 0.024 day(-1)) (p = 0.361). TRR > 0.05 day(-1) was seen in four chemoradiotherapy tumors (30.8%) and in six radiotherapy-alone tumors (15.0%) (p = 0.207), whereas TRR < 0.01 day(-1) was seen in no chemoradiotherapy tumors (0.0%) and in five radiotherapy-alone tumors (11.1%) (p = 0.180). TRR for tumors > 5.0 cm in diameter was greater with chemoradiotherapy (n=5) than with radiotherapy alone (n=12) (p = 0.065). CONCLUSION: Although the difference did not reach a statistically significant level, our TRR data suggest that concurrent chemotherapy heightens the radioresponse of large-size cervical cancer. PMID- 15786749 TI - CT-guided automated cutting needle biopsy by a combined method for accurate specific diagnosis of focal lung lesions. AB - PURPOSE: The purpose of our study was to evaluate a method of automated cutting needle biopsy (ACNB) that combines the use of a long-throw needle, higher mean number of needle passes, and tandem system, in terms of the accuracy of specific diagnosis of small and large lung lesions and the safety of the procedure. MATERIALS AND METHODS: Fifty-seven ACNBs were performed under computed tomography guidance using a tandem system with a 20-gauge and 18-gauge (through non-aerated lung) automated cutting needle with a throw length of 23 mm. We classified the nodules into 21 small nodules (< or =2 cm) and 36 large nodules (>2 cm). All ACNB diagnoses were divided into three groups: specific, non-specific, and false diagnoses. All of the complications were recorded. RESULTS: The mean number of ACNB specimens obtained was 2.0. Of the 35 ACNB procedures for malignant lesions, 33 yielded a specific malignant diagnosis (33/35, 94%). Of the 22 procedures for benign lesions, 17 gave a specific benign diagnosis (17/22, 77%). The diagnostic accuracy for small nodules was no lower than that for large nodules. Postbiopsy pneumothorax occurred in 18 patients (32%). CONCLUSION: The diagnostic accuracy of the combined method is as high for small lung nodules as for large ones. The procedure has high diagnostic accuracy for the subtypes of lung cancer and an acceptable complication rate. PMID- 15786750 TI - Relation between chromosomal aberrations and radiation dose during the process of TBI. AB - PURPOSE: The purposes of this study were (1) to consider the dose-response relationship in-vivo at high dose range by using the Colcemid method, which begins with predicting the risk of future late complication caused by a fixed dose of chromosomal aberration of peripheral lymphocytes after radiation emission and (2) to compare in-vivo and in-vitro dose-responses for chromosomal aberrations in lymphocytes of cancer patients undergoing total body irradiation (TBI). METHODS: Eight patients diagnosed with hematological malignancies entered this study. TBI planning with a 6 MV linear accelerator consisted of 4 Gy/2 fractions/day, and the total treatment dose was 12 Gy. RESULTS: At the observable dose range of up to 10 Gy, the unstable chromosomal aberrations of both dicentrics and fragments increased with the increment of irradiated dose, regardless of in-vivo or in-vitro irradiated samples. However, the average number of dicentrics and fragments obtained from in-vitro samples was found to be higher than that for in-vivo samples. CONCLUSION: This result strongly suggests that in vivo dose-response curves are necessary to estimate the absorbed dose in-vivo. In vivo dose-response curves obtained from cancer patients would be very important for standard curves for biodosimetry. PMID- 15786751 TI - High-spatial-resolution MR imaging of focal breast masses: interpretation model based on kinetic and morphological parameters. AB - PURPOSE: To assess a new interpretation model combining kinetic pattern and morphologic characteristics using high-spatial-resolution MR imaging. MATERIALS AND METHODS: Breast MR imaging was performed in 58 patients with 63 pathologically proved focal breast masses (49 malignant and 14 benign). MR imaging was performed on a 1.5-Tesla system using the volumetric interpolated breath-hold examination (VIBE) sequence. Morphological parameters were lesion shape, mass margin, rim enhancement (RE), enhancing internal septations, and internal signal on T2-weighted images. Lesion shape/margin was classified into four categories as follows: smooth (smooth/round or smooth/oval), lobulated (lobular shape), irregular (irregular margin or irregular shape), and spiculated (spiculated margin). RE was sub-classified as early RE at 60 sec and delayed RE at 4 min. Strongly high signal and the presence of internal black septations were evaluated over the entire lesion on T2-weighted images. The kinetic parameter was visually assessed as follows: washout, plateau, and progressive. RESULTS: The most frequent types of lesion shape/margin in the malignant lesions were irregular (47%) and spiculated (43%), whereas 13 of the benign lesions had smooth or lobular margins (93%). The most frequent characteristics in breast cancers with lobulated configuration were washout pattern (80%), whereas 78% of the lobulated benign lesions were negative for visual washout. The positive predictive value (PPV) was 98% (49/50). CONCLUSION: "Washout with enhancing septations" is thought be specific for carcinoma with a lobulated configuration (80%). A combination of morphological criteria (including lesion shape/margins, strongly high signal on T2-weighted images) and visual washout is useful for differentiating between benign and malignant lesions. PMID- 15786752 TI - Residual stenosis after conventional balloon angioplasty for hemodialysis shunt failure: treatment with metallic stent placement and post-balloon dilatation. AB - PURPOSE: The usefulness of metallic stent placement and post-balloon dilatation was investigated for patients with residual stenosis after conventional percutaneous transluminal balloon angioplasty (balloon PTA) of dialysis shunt vessels. MATERIALS AND METHODS: Among 92 patients who had received balloon PTA for dialysis shunt vessels, seven patients who showed a residual waist on the balloon even under maximum inflation were enrolled in this study. In these patients with residual stenosis after balloon PTA, we inserted a stent in the residual stenosis, and post-balloon dilatation was immediately applied using the same balloon catheter. RESULTS: After balloon PTA, the average percent diameter stenosis declined to 45.5 +/- 7.30%, and the stenosis was further improved to an average of 19.3 +/- 7.09% after the placement of a stent and the additional balloon PTA. The average percent diameter dilatation of the balloon before the placement of a stent was 65.8 +/- 12.7%, while the average dilatation increased to 84.1 +/- 8.96% after the placement of a stent. Dialysis became possible immediately after the procedure in all cases. CONCLUSION: Metallic stent placement and post-balloon dilatation was effective for patients with residual stenosis after conventional balloon PTA of dialysis shunt vessels. PMID- 15786753 TI - Phase shift due to inversion recovery process and chemical shift effect in inversion recovery prepared gradient-echo magnetic resonance imaging. AB - PURPOSE: The purpose of this study was to assess phase shift due to the inversion recovery process and the chemical shift effect in inversion recovery prepared gradient-echo magnetic resonance imaging. MATERIALS AND METHODS: The signal intensities of three phantoms, composed of physiologic saline solution with Gd DTPA, olive oil, and fat emulsion, were measured on a 1.5 Tesla unit. The sequence was single-shot two-dimensional. The parameters were as follows: TI 100 2500 msec at 100 msec intervals, TR 11.25 msec, TE 2.250 msec, 3.375 msec, and 4.500 msec. The k-space was filled with the centric view order. RESULTS: The signal intensity curves of the emulsion showed various patterns. At a TE of 2.250 msec, TI at the minimum value of signal intensity was 1500 msec. At a TE of 4.500 msec, TI at the minimum value was 500 msec. Up to a TI of 1000 msec, the signal intensity at a TE of 2.250 msec was stronger than that at a TE of 4.500 msec, and from a TI of 1000 msec, the signal intensity at a TE of 4.500 msec was stronger than that at a TE of 2.250 msec. CONCLUSION: We concluded that fat emulsion showed paradoxical phase shift. PMID- 15786754 TI - Multiphase contrast-enhanced CT of the liver with a multislice CT scanner: effects of iodine concentration and delivery rate. AB - PURPOSE: To determine whether or not high-concentration contrast material is useful in multiphase contrast-enhanced CT of the liver with a multislice CT scanner. MATERIALS AND METHODS: One hundred twenty-four examinations, in which first- and second-pass acquisitions (double arterial phase imaging) were performed during a single breath-hold followed by third-pass acquisition, were randomized into three protocols: contrast injection at 0.07 mL/kg body weight/sec over 30 sec at an iodine concentration of 300 mgI/mL in group 1, contrast injection at 0.06 mL/kg body weight/sec over 30 sec at an iodine concentration of 350 mgI/mL in group 2, and contrast injection at 0.07 mL/kg body weight/sec over 25.7 sec at an iodine concentration of 350 mgI/mL in group 3. Each group received an equivalent iodine dose per kg body weight (2.1 mL/kg of contrast material of 300 mgI/mL). Contrast enhancement in each acquisition was measured in the aorta, portal vein, and liver. RESULTS: No statistically significant differences were seen between groups 1 and 2 in any enhancement in any acquisition. In group 3, aortic enhancement in the first-pass acquisition was significantly more intense than in groups 1 and 2, while portal venous enhancement and hepatic enhancement were equivalent. CONCLUSION: Shortening the injection duration for a given iodine dose with high-concentration contrast material (group 3) can achieve improved arterial enhancement on arterial phase images. PMID- 15786755 TI - Ovarian fibroma (fibrothecoma) with extensive cystic degeneration: unusual MR imaging findings in two cases. AB - Two cases of ovarian fibroma (fibrothecoma) with extensive cystic degeneration are described. One case revealed a large multiloculated cystic mass with a small solid part. Each locule showed variable signal intensity on both T1- and T2 weighted magnetic resonance (MR) images. The other case revealed a parviloculated cystic mass with a small solid part. In both cases, the solid parts were located at the periphery of the mass and showed distinct low signal intensity on both T1- and T2-weighted MR images and slight enhancement. It should be noted that ovarian fibromas (fibrothecomas) with extensive cystic degeneration are a rare exception to the rule that solid components in a cystic adnexal mass imply malignancy. PMID- 15786756 TI - Ring enhancement on T1-weighted GRE images after ferucarbotran administration for hepatic metastasis: comparison with pathological findings: case report. AB - Hepatic metastases are generally known to be composed of three structures (central necrosis, viable tumor cells, and peritumoral changes such as edema or congestion). We experienced a case of hepatic metastasis that was examined with ferucarbotran-enhanced magnetic resonance (MR) imaging, dynamic MR imaging using Gd-DTPA, and CT during arterial portography (CTAP), and was confirmed pathologically. MR imaging of this case vividly described the three pathological structures of the hepatic metastasis. PMID- 15786757 TI - Geometrical verification system using Adobe Photoshop in radiotherapy. AB - PURPOSE: Adobe Photoshop is used worldwide and is useful for comparing portal films with simulation films. It is possible to scan images and then view them simultaneously with this software. The purpose of this study was to assess the accuracy of a geometrical verification system using Adobe Photoshop. MATERIALS AND METHODS: We prepared the following two conditions for verification. Under one condition, films were hanged on light boxes, and examiners measured distances between the isocenter on simulation films and that on portal films by adjusting the bony structures. Under the other condition, films were scanned into a computer and displayed using Adobe Photoshop, and examiners measured distances between the isocenter on simulation films and those on portal films by adjusting the bony structures. To obtain control data, lead balls were used as a fiducial point for matching the films accurately. The errors, defined as the differences between the control data and the measurement data, were assessed. RESULTS: Errors of the data obtained using Adobe Photoshop were significantly smaller than those of the data obtained from films on light boxes (p < 0.007). CONCLUSION: The geometrical verification system using Adobe Photoshop is available on any PC with this software and is useful for improving the accuracy of verification. PMID- 15786759 TI - [Terminology for lower urinary tract function]. PMID- 15786758 TI - [Urological historiography]. PMID- 15786760 TI - [Problem-based learning in urology training. The Faculty of Medicine of the Universidad de Castilla-La Mancha model]. AB - Ongoing changes in the social, economic, technological and scientific realms have generated new needs and led various organizations to suggest that educational institutions should reorient their educational strategies toward developing effective professionals with the skills to meet these needs. These "modern" strategies include problem-based learning, in which the student seeks and selects information, analyzes the data obtained, integrates both prior and newly acquired knowledge, and, finally, offers diagnostic and therapeutic options to resolve the problem posed, as would occur in professional practice. With this approach, prior skills and practical experience form the foundation of learning. Problem-based learning incorporates some aspects of cognitive psychology, a model that mainly centers on the nature of the knowledge structures found in active memory, the processes involved in information storage and retrieval and the various factors that activate these processes. At the Faculty of Medicine of the Universidad de Castilla-La Mancha, urology is part of a core subject (Medical and Surgical Pathology II) taught in the fifth year of coursework together with nephrology. Each course includes approximately 75 students, divided into five groups. The rotation lasts six weeks, with students spending a mean of two hours a day on theory (nephrology and/or urology) and the remaining time on rotations in the various activities: three weeks in nephrology and three weeks in urology. Upon completion of the rotation, the students write a combined theoretical examination with 100 multiple-choice questions (50 on urology) and take a practical skills examination. At the end of the course, another practical test consisting of an objective, structured clinical examination is taken, in which standard patients are used and the professor directly assesses the level of skills acquired with a "real" case. PMID- 15786761 TI - [Clinical applications of the comprehensive theory of urinary incontinence]. AB - Currently, urinary incontinence cannot be treated without taking into consideration the comprehensive theory of urinary incontinence. According to this theory, stress incontinence, urge incontinence and alterations in bladder emptying result from alterations in the elements of suburethral support, ligaments and muscles of the pelvis floor. Alterations in the forces applied by muscles and ligaments to the fascia adjacent to the vaginal wall and the urethra cause the vesical neck and the urethra to open or close. Alterations in the forces applied to the vagina can also cause premature activation of the micturition reflex, triggering involuntary contractions of the detrusor muscle. Knowledge of the anatomic substrate of the upper, intermediate and lower layers of the pelvic floor and of the main pubourethral, urethropelvic and uterosacral ligamentous elements is essential before any surgical approach of incontinence can be attempted. Depending on the location of the fascial or ligamental muscle lesion and of the sensitivity of the local nerve endings, stress incontinence, urge incontinence, altered bladder emptying or combinations of these conditions can develop. There are 6 basic defects which should be systematically investigated: defect in the suburethral support ("Hammock"), tethered vagina syndrome, distended pubourethral ligaments, distended uterosacral ligaments and support of the vaginal apex, lesions of vaginal insertion of pubococcygeal muscles, lesions of the striated muscles of the pelvic floor levator plate. PMID- 15786762 TI - [The mortality of prostate cancer in Andalusia. Contributions to mass screening]. AB - BACKGROUND: To describe the mortality of prostate cancer in Andalusia from 1975 to 2001. METHODS: Estimation of crude rates, age-adjusted rates, Potential Years of Life Lost (PYLL) adjusted rates, risk of death and the changes produced in the adjusted rates. RESULTS: Prostate cancer deaths rise from 407 to 767. Crude rates increased from 13.54 to 21.35 per 100,000 persons-years. However, the age adjusted rates showed a steady trend over the study period, with an annual percent change of -0.21%, and the premature mortality indicators declined. The cumulative risk of death increased with age, and presented an exponential increase after the age of 50 years, in both the periods, 1975-1979 as 1997-2001, with highest values in the second one. CONCLUSIONS: Prostate cancer mortality in Andalusia has increased in absolute values due to the ageing of the population. When adjusted for age, the mortality has remained stable, with a shift of deaths toward the more extreme age groups, as indicated by the decrease of premature deaths. The results of this study do not support the starting-up of mass screening programs. PMID- 15786763 TI - [Retrospective study of 130 patients with organconfined prostate cancer treated with brachytherapy]. AB - INTRODUCTION: The prostate brachytherapy with I 125 seeds has an indication in patients with organconfined prostate cancer. Our objective is to describe the population treated in our institution with permanent I125 seeds implants, the dosimetric characteristics of the technique and the preliminary results of our group-study in terms of evolution and toxicity. MATERIAL AND METHODS: Between May 2000 and March 2003, 130 patients with permanent implants of I125 seeds were treated. Beforehand we did prostate volumetric with transrectum prostate echography in order to assess the configuration of the implant, number of seeds and their place in the prostate with the objective to get a fine coverage of PTV (planet target volume). Stage distribution: 75.72% T1c; 24.28% T2a; Gleason<6, 94%. The PSA pretreatment average was 6.38 ng/ml. The average prostate volume was 30 cc. The 16.67% of the patients included had hormonal treatment previously to get the implants. The average age was 64 years. The characteristic techniques of the implants were: the average width of the needle as 24 (14-35) and the average of the seeds 76 (46-111). Finally the average activity was 0.39 mCi/seed, which means average total implant activity of 80 mCi. RESULTS: We analyzed 130 patients with average follow up 6 months. A 1 to 2 year surveillance was carried out on 98.9% and the global free disease surveillance (biochemic relapse) of 98.9% at the year and of the 87.8% at the end of the 2 years. The relapse in the low risk patients was 98.8% after the first year and 88.7% at the end of 2 year. On the contrary in the middle risk was of 100% and 83% respectively, although the amount of patients in significantly less. As a relevant acute secondary effects we found slight rectitys or GI (RTOG scale) in 1.4 and that needs synthomatic medication or GII (RTOG scale) in 0.8%. We found slide hematuria or GI (RTOG scale) in the 53% and other measures or GII (RTOG scale) in the 2.64% was needed. Finally we had to set a urinary prove for acute retention in 4.3%. CONCLUSION: The prostate brachyterapy is a complex procedure that needs a multidisciplinary team participation in order to be able to carry out. It avoids a long term hospitalization and allows for the patient to have daily activity within a short period of time. Despite the fact of the brief follow-up, the results over biochemical relapse and toxicity were similar to the ones in the literature. Tolerance to the implant was good. It would necessary a longer follow-up in order to be able to come to long term conclusions. PMID- 15786764 TI - [Evolution of the clinical, echographic and pathological characteristics of patients with indication of transrectal ultrasound and prostate biopsy between 1994 and 2003]. AB - OBJECTIVE: The aim of this paper is to evaluate the evolution in the clinical, echographic and pathological characteristics of patients undergoing transrectal ultrasound and prostate biopsy over a 10 year period (1994-2003). MATERIAL AND METHODS: Patients undergoing a first biopsy of the prostate between 1994-2003 were studied retrospectively. Since the biopsy criteria have changed several times over this period, results were analysed for four different periods: 1994 1997, 1998, 1999-2003 (the latter was further divided into two periods--1999-2001 and 2002-2003--to observe the temporal evolution with the same biopsy criteria). Patients were assigned to risk groups according to D'Amico. RESULTS: We observed that individuals referred for biopsy and those diagnosed with prostate cancer (PC) had a lower mean age (p=0.0001 and p=0.01), there were more patients with a PSA from 4.1-10 ng/ml (p=0.0001 and p=0.0001), more patients had no significant DRE findings (p=0.0001 and p=0.0001) or ultrasound findings (p=0.0001 and p=0.0001). The incidence of cancer diagnosis has decreased from 39.4% to 34.3% (p=0.03). There was an increased incidence of score 7, at the expense of a decline in scores 5-6 and 8-10 and disappearance of score 2-4 (p=0.0001). Patients assigned to the low risk group increased from 9% to 18.1%, those assigned to the intermediate risk group from 20.2% to 43.1% and there was a reduction in those assigned to the high risk group from 70.7% to 38.7% (p=0.0001). Analysis of the final two time periods revealed that the evolution of the series remained the same except for a decrease in age at biopsy and diagnosis of cancer. CONCLUSIONS: Nowadays, the risk factors of patients with an indication of biopsy have less weight than ten years ago. We currently diagnose patients with PC with more favourable prognostic factors. However, the price we pay for this earlier diagnosis is reflected in a less effective biopsy, a larger proportion of the population without PC having to experience the physical complications and psychological stress of a biopsy, a greater number of patients having to undergo a second biopsy and, therefore, a greater and more costly use of resources to diagnose PC. PMID- 15786765 TI - [Expression of the cerbB-2 (HER-2/neu) oncoprotein in prostatic adenocarcinoma]. AB - OBJECTIVES: Our aim is to determine the expression of the cerbB-2 oncoprotein in prostate cancers using an immunohistochemistry staining and to compare these results with several clinical and histological prognostic factors. METHODS: An immunohistochemical staining using the cerbB-2 monoclonal antibody (Dako) was performed in 32 radical prostatectomy specimens diagnosed of adenocarcinoma. The intensity of cerbB-2 expression was evaluated with a scale that variated from 0 (no staining) to 3+ (strong complete membrane staining) according to published guidelines. Association of cerbB-2 index immunoreactivity with clinical and histological prognostic factors was examined. RESULTS: Definite positive membranous staining was detected in 14 of 32 neoplastic cases (44%). Such overexpression was correlated with higher Gleason grade (p=0.04) and higher stage of disease (p=0.038). CONCLUSIONS: 1) This study shows that 44% of all prostate cancer express the cerbB-2 oncoprotein with immunohistochemical technique. 2) These findings suggest that is necessary to standardize the immunohistochemical staining procedure with cerbB-2 in prostate adenocarcinoma. 3) The level of cerbB 2 expression was correlated with Gleason grade and clinical stage. PMID- 15786766 TI - [Localized prostate cancer treatment in renal transplant patient with high intensity focalized ultrasound (HIFU)]. AB - We report a 62 years old kidney transplant (KT) patient who was diagnosed of localized prostatic cancer (PC) after 6 years of the implant. Transrectal prostatic High Intensity Focused Ultrasound (HIFU) was applied. Results have been satisfactory, achieving pathologic and biochemical success. The discharge was completed at 24 hs, the morbidity was minimal. We have not found any reference in the literature on the appliance of HIFU in PC KT patients. We think that HIFU may represent a good alternative for these patients. PMID- 15786767 TI - [Evaluation and prognostic of cystic renal tumors]. AB - OBJECTIVE: To evaluate the clinical and pathological characteristics of cystic renal tumors in our center. MATERIAL AND METHODS: A retrospective review of 239 nephrectomies is performed comparing the clinical and pathological variables of cystic tumors with those of solid renal tumors. Survival outcomes are analyzed in both groups. RESULTS: Our experience shows that cystic renal tumors behave like solid renal tumors, with no differences in survival shown. The variables studied show statistically significant differences in histological grade and number of tumors, with cystic tumors having a lower histological grade and being more often multiple in number. CONCLUSIONS: The behavior of cystic renal tumors is no different than that of solid renal tumors. PMID- 15786768 TI - [Age's influence in superficial bladder cancer's behaviour]. AB - INTRODUCTION: Transitional bladder cancer is infrequent in younger people. The tumor's aggressiveness in this population is subject of discussion. We present our experience. MATERIAL & METHOD: 551 primary tumors, with at least one year of follow-up. Group A (less than 41 years old), Group B (bigger than 40 years old). We compare, stage, grade, lymphatic permeation, multiplicity, recurrence, progression, sex, T.L.E. (time free of disease), volume. We do uni and multivariate analysis. RESULTS: 17 tumors in younger than 41 years (2.1%). The recurrence rate is 11.8% in Group A and 36% in Group B (p .041). Progression rate: 0% in A and 4.1% in B (p .253). In Group A, T1 tumors (13.3% vs. 39%), G2 tumors (0% vs. 36.7%); withouts in lymphatic permeation (0% vs. 35%) and solitary tumors (13.3% vs. 35.3%) have statistically less recurrence. In univariate analysis we appreciate statistical differences in relation with grade (p .002) and sex (p .011). In multivariate analysis, stage, "group of age" and prophylactic treatment are independent variables for recurrence, and grade for progression. CONCLUSIONS: In younger than 41 years old: Less pathologic aggressivity (0% G3); Group of age is independent prognostic variable for recurrence, but not p for progression, despite a "less aggressive behaviour": 11.8% recurrence, 0% progression; T.L.E. bigger (one year more); Higher incidence in women (41% vs. 11%); Less recurrence in T1 tumors (13.3% vs. 39%), G2 tumors (0% vs. 36.7%) and solitary tumors (13.3% vs. 35.32%). PMID- 15786769 TI - [Transobturator correction of cystocele]. AB - OBJECTIVE: We present our initial experience with the Perigee system for anterior vaginal wall prolapses repair. MATERIAL AND METHOD: 15 patients with anterior vaginal wall prolapse (mean age: 62 years old) underwent implanted with Perigee system which is composed by a mesh for correcting central defect and four self attached horns for correcting lateral defect. The implanted procedure consist of four steps: 1) lateral vaginal wall dissection as far as isquiopubic branch; 2) performance of two upper marks at genitofemoral fold and two lower marks at 2 cm laterally and 3 cm lower; 3) insertion of the upper needles parallel to the isquiopubic branch and connection with the mesh's "arms" plus insertion of the lower needles vertically and connection with mesh's "legs"; 4) to adjust the mesh free tension. The system POP-Q was used as an objective measure of correction rate. RESULTS: Preoperatively, the point Aa was 0.09. After 6 weeks and three months postoperatively was -3.00 and -2.73 respectively. Preoperatively, the point Ba was 1.73. After 6 weeks and three months postoperatively was -2.82 and 2.82 respectively. No major complications were presented. No vascular damage or significant bleeding was observed. CONCLUSION: The transobturator correction of cystocele is an attractive alternative. The initial good outcome may be expected to be long lasting. PMID- 15786770 TI - [Incomplete urethral duplication in a male. A case report]. AB - Urethral duplication is a rare congenital anomaly affecting mainly males and being usually diagnosed during paedriatric age. We report a 20 year old male complaining of double urethral meatus with double urinary stream. Physical examination confirmed and additional hypospadic meatus below a normally placed urethral meatus. Retrograde urethro-cystography and voiding cysto-urethrograms showed two distinct urethras originating from a common bladder neck and the diagnosis of Effmann type IIA2 incomplete urethral duplication was made. No treatment was felt to be applied after associated anomalies were ruled out. PMID- 15786771 TI - [Urologic findings in pseudoxanthoma elasticum: report one case]. AB - CASE REPORT: A 21-year-old woman presented with retinal angioid streaks and yellowish streak skin abnormalities in neck and axillary folds. Skin biopsy showed bluish-gray tangled masses of calcified elastic fibers in the mid- to lower dermis suggestive of Pseudoxanthoma elasticum (PXE). She consulted in Urology Department for lower urinary tract infection. Renal ultrasonography revealed multiple highly reflective foci in the corticomedullary junction. COMMENT: Pseudoxanthoma elasticum is a rare genetic disorder characterised by fragmentation and calcification of elastic fibers in the skin and media of arteries. Frequent manifestations include hypertension, angina pectoris, transient cerebral ischemic attacks, intermittent claudication, upper gastrointestinal bleeding, retinal angioid streaks and thickened skin. A characteristic appearance of highly reflective foci in the renal parenchyma have been reported in patients with PXE, but it's not specific for this syndrome. However, the presence of this structural pattern in a young patient with dermatological abnormalities should lead to the consideration of PXE in the differential diagnosis list. Besides ultrasonography findings, only a ureteral disruption case in a patient underwent ureteroscopic manipulation has been described to our knowledge. In our case, the true significance of these disease in recurrent urinary tract infection is uncertain. PMID- 15786772 TI - [Prostatic abscess: report of a case and review of the literature]. AB - Prostatic abscess is an uncommon entity nowadays due to widely use of antibiotics, however, in some patients, because of the seriousness, a quick diagnosis and interventionist treatment is required. We report a case of a 78 year-old male patient who had a torpid evolution and needed an urgent prostatectomy. We review ethiopathogenic factors, clinical findings, diagnosis and treatment of this uncommon entity. PMID- 15786773 TI - [Intraprostatic embolization of cholesterol crystals]. AB - The cholesterol embolism syndrome (CES) is an unusual disease that carries a high mortality rate. Finding intraprostatic cholesterol crystal embolization as the result of transrectal prostate biopsy in a patient with several risk factors for atherosclerosis, should alert the urologist to the possibility of CES existence. PMID- 15786775 TI - [Squamous cell carcinoma in bladder extrophy]. AB - Bladder extrophy is a rare congenital malformation that nowadays is surgically corrected in neonatal period. We present a case report of a 71-year-old male with a verrucous squamous cell carcinoma arising in a classical uncorrected form of bladder extrophy. PMID- 15786774 TI - [Non obstructive retrocaval ureter]. AB - We report a case on circumcavum ureter without obstruction or type two, according the Batenson and Atkinson's classification, in a forty-nine year old man. It was diagnosticated for a gross haematuria and renoureteral pain, because of a simultaneous urinary tract infection. We emphasize its absence of the typical morphology and obstruction signs. After twelve months the patient is still asymptomatic, without any medical or surgical treatment. PMID- 15786776 TI - [Considerations about angiolymphoid hyperplasia with eosinophilia (ALHE) with regard to a case localized in the penis]. AB - We report the case of a 27 year old man who presented a nodule in the ventral face of the penis, which increased in volume and turned painful during erection so a vascular lesion was suspected, later confirmed with a Doppler study. A complete excision of the lesion was carried out with local anaesthesia. The pathological specimen was informed as "angiolimphoid hyperplasia with eosinophilia" in its intravascular form. This is the third communication of this kind of lesion in the penis but no other adopted the intravascular variant like the present case. We discuss about this exceptional entity and make review of the most relevant literature. PMID- 15786777 TI - [Reflections on the Hispano-American integration of Actas Urologicas Espanolas]. PMID- 15786778 TI - [Entero-vesical fistula in Behcet's disease]. PMID- 15786779 TI - [Radio-opaque lithiasis and indinavir]. PMID- 15786780 TI - Some thoughts on change. PMID- 15786781 TI - The top 10 reasons elective cesarean section should be on decline. PMID- 15786782 TI - Predicting pre-eclampsia. Substance in urine signals likelihood of high-risk development. PMID- 15786783 TI - Healthy hearts for women. Making a difference with AWHONN's evidence-based guideline. PMID- 15786784 TI - ACOG issuse hormone therapy guidelines. Experts expand estrogen advice; renounce herbs for hot flashes. PMID- 15786785 TI - Keeping your practice cutting-edge. Why using research on practice matters. PMID- 15786786 TI - Providing evidence-based care. You can understand research and use it in practice! PMID- 15786787 TI - Tocolytics for pre-term labor. What nurses need to know. PMID- 15786788 TI - Finding buried treasure in unit log books. Data mining for gold. PMID- 15786789 TI - A new generation of women's health services. Growing service lines beyond reproductive health. PMID- 15786790 TI - Exploring breast cancer therapies. Comparing aromatase inhibitors to current therapies. PMID- 15786792 TI - Cardiovascular nursing research--whence from here? PMID- 15786791 TI - A perinatal CNS? What is that? PMID- 15786793 TI - Changes in cardiac surgery. AB - The specific changes of same-day admission, choice of bypass conduit, use of cardiopulmonary bypass and rapid recovery have been highlighted to reflect current changes in cardiac surgical care. Practice guidelines with respect to symptom management of postoperative pain, nausea and atrial fibrillation are future topics to be discussed reflecting additional practice changes in cardiac surgery. PMID- 15786794 TI - A qualitative analysis of the experience of uncertainty while awaiting coronary artery bypass surgery. AB - The aim of this article is to present a qualitative examination of the phenomenon of uncertainty in patients awaiting CABG surgery. Cardiac symptoms are presented as an antecedent to uncertainty and both positive and negative outcomes of uncertainty are discussed. Semi-structured interview questions, based on Mishel's Uncertainty in Illness Theory, were developed to investigate the experience of waiting for CABG surgery and identify what patients were doing: to make their cardiac symptoms more manageable, to relieve their own anxiety while waiting, and to identify why patients felt having CABG surgery would benefit them. Telephone interviews were conducted with a self-selected sample of 25 participants. The qualitative results presented are part of a larger multimethod study examining the psychosomatic experience of waiting for CABG surgery. Data collected from telephone interviews were analyzed using content analysis and constant comparison techniques. Content analysis identified three conceptual categories: i) taking responsibility; ii) getting my life back; and iii) getting it over with. Strategies associated with each of these categories, the consequences of the strategies, and factors that facilitate or constrain their use were also identified. Participants were actively trying to lessen the impact of their cardiac symptoms and were very aware of their own bodies and what actions would exacerbate symptoms or relieve symptoms if they occurred. Participants envisioned physical and psychological improvements for post-CABG surgery. Participants also identified actions they took to limit their anxiety while waiting. Anxiety levels were influenced by family members as well as stories they had heard from friends, family, or acquaintances who had also experienced CABG. This study concluded that the lengthy waits experienced by some patients can create significant psychological disturbance including anxiety and uncertainty about the future. In addition, patient symptom status requires attentive monitoring. PMID- 15786795 TI - Increasing patient-initiation of cardiac rehabilitation referral in female percutaneous coronary intervention patients. AB - Substantial health risks continue following coronary revascularization, and cardiac rehabilitation (CR) plays a large role in enhancing prognosis. However, fewer women than men are referred to and participate in CR. This study prospectively evaluates a psycho-educational intervention designed to increase patient-initiation of CR referral discussion with their physician, It was hypothesized that an educational brochure along with a motivational interview would facilitate the discussion of CR between physicians and female patients, ultimately resulting in increased referral of women to CR when compared to a control group. Eighty female percutaneous coronary intervention patients were recruited from two acute care sites in Toronto, Ontario. Eight weeks later, a second research assistant blind to condition telephoned participants to assess aftercare and CR referral processes. Results showed that patients in the experimental group more often discussed CR with a health care provider (chi2(1)=5.99, p=.01), and there was a trend toward increased referral when compared to control patients (chi2(1)=2.83, p=.09). Gaps in secondary prevention are noted, and areas for improved referral of women patients to CR are discussed. PMID- 15786796 TI - Gender differences in functional capacity following myocardial infarction: an exploratory study. AB - This study examined functional capacity and hemodynamic responses to exercise in men and women during early recovery after myocardial infarction (MI). Participants were tested at hospital discharge and six weeks following discharge. At each testing time, they completed the Beck Depression Inventory and a symptom limited graded exercise test (SL-GXT). Functional capacity, expressed in metabolic equivalents (METs), was estimated from performance on the SL-GXT. Hemodynamic response measures included heart rate (HR), systolic blood pressure (SBP). and rate-pressure product (RPP). Both men and women demonstrated significant increases in functional capacity from discharge to six weeks later although the levels were lower for women (M = 2.8 METs) than men (M = 7.3 METs). This indicates a need for gender-sensitive counselling for unsupervised exercise rehabilitation during early recovery following MI. PMID- 15786797 TI - Cultural variants of caregiving or the culture of caregiving. AB - Research has shown that ethnic variations in caregiving exist. However, less attention has been given to the common experience of caregiving, across ethnic group affiliation. Semi-structured interviews with African American, Caucasian American, and Latino caregivers were conducted to understand the common experience of caregiving. Four major themes that characterized the caregiving experience across the three ethnic groups were identified. Participants described a tradition of family care, prevalent across the three ethnic groups. Familial conflicts, caregiving dissonance, and low levels of use of formal services also were prominent characteristics of the caregiving experience across the three ethnic groups. Results support the existence of a culture of caregiving prevalent across ethnic group affiliation. PMID- 15786798 TI - Factors influencing exercise participation and quality of life among elderly Korean Americans. AB - The purpose of this study was to identify factors influencing exercise participation and quality of life among elderly Korean Americans. This study used a focus group design. Elderly Korean Americans age 60 and older residing in Seattle, Washington were recruited from a senior house, which serves low-income elderly. Thirteen older adults participated in a focus group discussion. Data analysis used content analysis. Factors influencing exercise participation included health status, advancing age, cultural self-consciousness, and lack of transportation. Factors influencing quality of life included health status, socioeconomic isolation, and spirituality. This study provides useful knowledge for designing future health promotion programs for this cultural minority group. PMID- 15786799 TI - New Mexico nurses' cultural self-efficacy: a pilot study. AB - Health care providers must develop a level of comfort in caring for patients/clients from cultures other than their own. A pilot study of 15 registered nurses caring for multiethnic patients in New Mexico was conducted to determine their level of self-efficacy, to test the Cultural Self-Efficacy Scale, and to determine the feasibility of a larger study. The participants were moderately confident with their knowledge of cultural concepts, cultural nursing skills, and three of the five ethnic groups' cultural life patterns. Although no conclusions can be drawn due to the small convenience sample, this study should lead to larger, more rigorous studies. PMID- 15786800 TI - Single embryonic stem cell-derived embryoid bodies for gene screening. PMID- 15786801 TI - Semiautomatic detection of DNA methylation at CpG islands. PMID- 15786802 TI - GST fusion vector with caspase-6 cleavage site for removal of fusion tag during column purification. PMID- 15786803 TI - Method for assigning double-stranded RNA structures. PMID- 15786804 TI - Extending AFLP sequences by long template PCR amplification of a fungal genomic library. PMID- 15786805 TI - Recombinational cloning vectors for regulated expression in Saccharomyces cerevisiae. PMID- 15786806 TI - Translation toeprinting assays using fluorescently labeled primers and capillary electrophoresis. AB - The protocol described here is an adapted version of the toeprinting assay in which the oligonucleotide used to prime the reverse transcription step is labeled with a fluorescent dye instead of 32P. By using a fluorescent dye, the results of the assay are obtained within one hour by direct electrophoresis of the samples on an automated sequencing machine. This eliminates the need to cast and run polyacrylamide gels and to wait to expose dried gels. We show that an identical toeprint was found for the chloramphenicol acetyltransferase transcript using this nonradioactive method, which is in agreement with the previously published 32P-labeled method. Furthermore, in addition to being a faster and safer method, a larger region of sequence can be analyzed with one primer in a single experiment. PMID- 15786807 TI - Strategy for chromosomal gene targeting in RecA-deficient Escherichia coli strains. AB - Reengineering DNA by homologous recombination in Escherichia coli often depends on helper functions provided on a temporarily introduced replicon that is subsequently cured from the cells. The suicide vector pKSS offers a new curing strategy. pKSS specifies a variant of phenylalanyl-transfer RNA (tRNA) synthetase conferring relaxed substrate specificity towards phenylalanine analogs that results in their lethal incorporation into cellular proteins. Consequently, the presence of p-chlorophenylalanine selects for strains that have lost pKSS. This principle, in conjunction with a plasmid-borne recA gene, was exploited for targeted chromosomal mutagenesis by double homologous recombination in RecA negative E. coli strains. Gene replacement with a kanamycin-resistance cassette was possible in a single step by plating on kanamycin and p-chlorophenylalanine agar plates and incubating at 37 degrees C. The presence of the correct chromosomal mutation and the absence of the plasmid were established by several control experiments. A simple screen confirmed the desired resistance phenotype in 44% of the initially selected clones, and 75% of these had the correct genotype. PMID- 15786809 TI - Disposable microfluidic devices: fabrication, function, and application. AB - This review article describes recent developments in microfluidics, with special emphasis on disposable plastic devices. Included is an overview of the common methods used in the fabrication of polymer microfluidic systems, including replica and injection molding, embossing, and laser ablation. Also described are the different methods by which on-chip operations--such as the pumping and valving of fluid flow, the mixing of different reagents, and the separation and detection of different chemical species--have been implemented in a microfluidic format. Finally, a few select biotechnological applications of microfluidics are presented to illustrate both the utility of this technology and its potential for development in the future. PMID- 15786810 TI - CDNA library construction from a small amount of RNA: adaptor-ligation approach for two-round cRNA amplification using T7 and SP6 RNA polymerases. AB - In this study, we developed a method that allows cDNA library construction from a small amount of RNA without causing serious size bias in the resulting cDNA population. For this purpose, we adopted two-round cRNA amplification by T7 and SP6 RNA polymerases. The first-round cDNAs, flanked by the promoter sequences of T7 and SP6 RNA polymerases, were synthesized from 1 microg total RNA and then subjected to two rounds of cRNA amplification. Comparison of the sizes of the first-round and the second-round cRNAs indicated that the size-bias effect of the second-round cRNA synthesis was not serious. The resultant double-stranded cDNAs were cloned into a plasmid by in vitro lambda phage recombination with an efficiency of 1.2 x 10(11) colony-forming unit/microgram of starting total RNA. Characterization of the resultant cDNA library in terms of the insert size, clone redundancy, and integrity of 3' ends of cDNAs indicated that the amplified library was comparable to a library constructed by a conventional method, although large cDNAs tend to be slightly truncated in the amplified library. This method enables the construction of a library from a small amount of RNA, and calculations suggest that the strategy would be efficient enough to use even a single cell as starting material. PMID- 15786808 TI - Quantitative imaging of protein interactions in the cell nucleus. AB - Over the past decade, genetically encoded fluorescent proteins have become widely used as noninvasive markers in living cells. The development of fluorescent proteins, coupled with advances in digital imaging, has led to the rapid evolution of live-cell imaging methods. These approaches are being applied to address biological questions of the recruitment, co-localization, and interactions of specific proteins within particular subcellular compartments. In the wake of this rapid progress, however, come important issues associated with the acquisition and analysis of ever larger and more complex digital imaging data sets. Using protein localization in the mammalian cell nucleus as an example, we will review some recent developments in the application of quantitative imaging to analyze subcellular distribution and co-localization of proteins in populations of living cells. In this report, we review the principles of acquiring fluorescence resonance energy transfer (FRET) microscopy measurements to define the spatial relationships between proteins. We then discuss how fluorescence lifetime imaging microscopy (FLIM) provides a method that is independent of intensity-based measurements to detect localized protein interactions with spatial resolution. Finally, we consider potential problems associated with the expression of proteins fused to fluorescent proteins for FRET based measurements from living cells. PMID- 15786811 TI - Diagnostic plots for detecting outlying slides in a cDNA microarray experiment. AB - Different sources of systematic and random error variations are often observed in cDNA microarray experiments. A simple scatter plot is commonly used to examine outlying slides that have unusual expression patterns or larger variability than other slides. These outlying slides tend to have large impacts on the subsequent analyses, such as identification of differentially expressed genes and clustering analysis. However, it is difficult to select outlying slides rigorously and consistently based on subjective human pattern recognition on their scatter plots. A graphical method and a rigorous diagnostic measure are proposed to detect outlying slides. The proposed graphical method is easy to implement and shown to be quite effective in detecting outlying slides in real microarray data sets. This diagnostic measure is also informative to compare variability among slides. Two cDNA microarray data sets are carefully examined to illustrate the proposed approach. A 3840-gene microarray experiment for neuronal differentiation of cortical stem cells and a 2076-gene microarray experiment for anticancer compound time-course expression of the NCI-60 cancer cell lines. PMID- 15786812 TI - The Shipman inquiry. PMID- 15786813 TI - The doctor-nurse relationship in the operating theatre. AB - This article examines the organisational culture in theatres, specifically the doctor-nurse relationship, based on the literature and the experience of one author (Anika Reynolds) on placement in theatres. The initial motivation for the study was noticing how well these doctors and nurses got on in a friendly, informal and efficient atmosphere. This relationship between the multidisciplinary team was especially surprising when compared with the experience of hospital wards. Why did such a difference exist? PMID- 15786814 TI - European Working Time Regulations. Implications for practice. AB - The European Working Time Regulations (EWTR) have implications for all NHS and independent sector employees. The aim of the EWTR is to protect employees' health and safety by encouraging improvements in the working environment. PMID- 15786815 TI - Eye surgery on the Thai Burma border. AB - Civil war and ethnic conflict are two of the intractable problems facing the world today, and they are followed by refugees and displaced people almost as inevitably as night follows day. This article looks at what has been achieved by two British ophthalmologists working with the local people in just one of these situations. PMID- 15786816 TI - How safe is your endoscope disinfectant? AB - Glutaraldehyde may have been virtually eliminated from UK hospitals as a flexible endoscope disinfectant, but in many cases it has been replaced with alternatives that may not be as safe as was first thought. This article highlights the need for healthcare professionals and hospitals to take urgent action to review the way these newer disinfectants are used and handled. PMID- 15786817 TI - The link between circulating markers of endothelial function and proteinuria in patients with primary glomerulonephritis. AB - INTRODUCTION: It is well established that there is an increase in the incidence of cardiovascular mortality in patients with proteinuric renal disease. The magnitude of the increase in risk is unlikely to be explained by traditional risk factors for cardiovascular disease alone. Proteinuria itself may constitute an additional risk factor, and proteinuric patients are known to have a degree of endothelial dysfunction. The nature of this relationship between proteinuria and endothelial function is the subject of intense investigation. AIM: The aim of this study was to examine the relationship between proteinuria and endothelial dysfunction, as reflected by serum von Willebrand factor (vWF), and the soluble cell adhesion molecules VCAM and ICAM, in patients with primary glomerulonephritis (GN). A secondary aim was to discern whether any relationship could be explained by renal function, lipid profile, inflammation or blood pressure. METHODS: A cross-sectional study was undertaken in consecutive patients attending a general nephrology clinic with biopsy-proven primary GN. Patients with end-stage renal disease (ESRD), those on immunosuppressive drugs, or with intercurrent infective illnesses were excluded. Blood pressure and body mass index were recorded. Routine lab assays were undertaken for serum creatinine, lipid profile, and 24-hour urinary protein (U(Prot)). Additional serum samples were stored at -80 degrees C for subsequent measurement of vWF, VCAM, ICAM and sensitive C reactive protein (sCRP). RESULTS: Data were collected from 129 (86 male) patients. Mean (standard deviation) estimated creatinine clearance was 64 (32) ml/min, and median (interquartile range) 24-hour proteinuria was 1.1 (0.22 2.9) g. Mean vWF was 173 (68) IU/dl, median VCAM, ICAM and sCRP were 594 (410 708) ng/ml, 235 (208-286) ng/ml, and 2.33 (0.83-5.68) mg/l, respectively. There was a significant positive correlation between vWF and U(Prot) (Spearman rank correlation, r = 0.41, p < 0.001). When split into tertiles, according to U(Prot) (0-500 mg, 500-2000 mg, and > 2000 mg), there was a significant, stepwise increase in mean vWF (p < 0.001), log VCAM (p < 0.001), and log ICAM (p = 0.002). On multivariate analysis with vWF as the continuous dependent variable, U(Prot), age, total cholesterol and sCRP were the only significantly independent correlates (model-adjusted R2 = 33%). CONCLUSION: In patients with primary GN, there is a significant association between endothelial activation as reflected by vWF, VCAM, or ICAM and increasing proteinuria. Elevations in vWF, as well as being related to classical risk factors, are associated with increases in total proteinuria and low-grade inflammation. Thus, future prospective studies should examine the extent to which vWF and other circulating markers of endothelial activation predict coronary heart disease risk in patients with proteinuric renal disease. PMID- 15786818 TI - Serum potassium, cardiovascular risk, and effects of an ACE inhibitor: results of the HOPE study. AB - BACKGROUND: Both hyper- and hypokalemia increase cardiovascular risk. Modest hyperkalemia is common with angiotensin-converting enzyme inhibition. We studied post-hoc the association of an initial, on-treatment serum potassium measurement with subsequent cardiovascular outcomes over 4.5 years in 9297 individuals at high cardiovascular risk, randomized to an ACE inhibitor or to placebo. METHODS: Post-hoc analysis of cardiovascular outcomes, as related to serum potassium levels, in the HOPE (Heart Outcomes and Prevention Evaluation) study which compared ramipril to placebo, and included 692 patients with a serum potassium level >5.0 mM and 137 with a serum potassium level <3.5 mM, defined as hyper- and hypokalemia, respectively. Serum potassium was measured 1 month after start of randomized treatment. RESULTS: With hyperkalemia, the primary event rate was unchanged compared to normokalemia (15.5 vs 15.7%, p > 0.4, respectively), with hypokalemia, the primary event rate was higher (22.6% vs 15.5%, respectively, p = 0.023). The hazard ratio for the primary outcome associated with this initial hypokalemia was 1.44 (1.00-2.06) on multivariate analysis. The combined primary outcome (myocardial infarction, cardiovascular death, stroke) was not different throughout deciles of serum potassium but the lowest and highest deciles included many with normokalemia. Randomized treatment was withheld because of hyperkalemia in 8 and 6 people allocated to ramipril and placebo, respectively. The benefit of ramipril on cardiovascular outcomes was independent of serum potassium, but ramipril reduced hypokalemia in the entire cohort (1.15 vs 1.86% with placebo, p = 0.005), particularly in those participants on diuretics (3.8% vs 6.5%, p = 0.07). CONCLUSIONS: In patients at high cardiovascular risk, modest hypokalemia predicts a less favorable outcome while modest hyperkalemia does not. Ramipril reduces hypokalemia and decreases risk. PMID- 15786819 TI - Risk factors determining active urinary stone formation in patients with urolithiasis. AB - AIMS: The goal of clinical and metabolic evaluation of patients with urinary stones is to identify patients at high risk for recurrent stone formation and as such, to allow the practitioner to suggest preventive therapies. However, knowledge about risk factors for active stone formation in patients with urolithiasis is limited. This study was initiated to assess the significance of several metabolic and clinical parameters for the determination of the risk of active stone formation. METHODS: Study in 320 consecutive outpatients referred to our clinic for metabolic analysis. Clinical and metabolic parameters were determined by standardized procedures of questionnaires, serum biochemical profiles and urinalysis. RESULTS: In 21.5% of 284 patients with complete data stone formation was active. Hypercalciuria, hypocitraturia and urinary tract infections had odds ratios for active stone formation above 2.5, whereas the odds ratio of a positive family history was 0.38. Hyperuricosuria, hyperoxaluria and a low urinary volume did not influence the risk for active stone formation. CONCLUSION: The risk profile for active stone formation differs from the risk profile for urolithiasis in general. Metabolic evaluation and determination of those risk factors in patients with urolithiasis might improve the estimation of the risk of future stone formation. PMID- 15786820 TI - Compartment-specific quantitative gene expression analysis after laser microdissection from archival renal allograft biopsies. AB - BACKGROUND: Various immunological and non-immunological pathomechanisms are responsible for the cellular damage in renal allografts. Since the kidney is an anatomically complex organ with functional and morphological heterogeneous compartments (interstitium, tubuli, vessels, glomeruli), the local response to injury maybe variable, therefore, the identification of local pathomechanisms is important. AIM: To elucidate any discrepancies in quantitative mRNA expression profiles between a total specimen analysis and a cell-specific evaluation after laser microdissection. METHODS: Real-time RT-PCR was performed for complement component C3 and heme oxygenase-1 (HO-1) genes compared to the housekeeping gene beta-actin using whole section RNA extracted from formalin-fixed and paraffin embedded archival material of 16 explanted, rejected renal allografts. Ten non transplant nephrectomies served as controls. For five cases from each group, five different compartments of the organs (interstitium, proximal tubuli, distal tubuli, vessels, glomeruli) were microdissected and quantitative analysis for C3 and HO-1 was performed identically. RESULTS: Whole section mRNA expression analysis: the data showed a constant expression of the housekeeping gene beta actin, a 7-fold increased expression of C3 and a 3-fold decreased expression of HO-1 in the allograft group as compared to the control group. mRNA expression results from microdissected compartments: in the control group, C3 and HO-1 expression could only be detected in the proximal tubuli of all cases whereas all five compartments analyzed from the rejecting kidneys showed expression of the two genes. In the allografts, expression levels of the investigated genes varied considerably not only among the different compartments but between individual cases as well. CONCLUSION: Laser microdissection combined with real-time RT-PCR is a feasible approach for retrospective quantitative gene expression analysis in formalin-fixed and paraffin-embedded renal allograft specimens. As shown for C3 and HO-1, cell-specific expression patterns ofpathogenetically relevant genes vary considerably between individual cases. A close correlation of morphology and cell-specific gene expression analysis will contribute to the elucidation of the complex pathogenesis of chronic renal allograft nephropathy. PMID- 15786821 TI - Impact of nocturnal home hemodialysis on anemia management in patients with end stage renal disease. AB - AIM: Anemia is adversely associated with poor uremia control and is an established cardiovascular risk factor in patients with end-stage renal disease (ESRD). Nocturnal home hemodialysis (NHD) is a novel form of renal replacement therapy that offers superior clearance of uremic solutes and improvements in several cardiovascular outcome parameters. We conducted a retrospective cohort study to test the hypotheses that augmenting the dose and frequency of dialysis by NHD would improve hemoglobin (Hb) concentrations and decrease requirement of erythropoietin (EPO) in ESRD patients. METHODS: In 63 patients (mean age: 46 +/- 2 years) receiving NHD (mean duration: 2.1 +/- 0.2 years), Hb, EPO dose, iron saturation, ferritin were determined before and at six monthly repeated intervals after conversion to NHD. For comparison, 32 ESRD patients (mean age: 57 +/- 3 years) who remained on self-care conventional hemodialysis (CHD) were also studied. RESULTS: There were no differences in baseline Hb concentrations, iron saturation, ferritin, or EPO dose between the two cohorts. After transfer from CHD to NHD, there were significant improvements in Hb concentrations (from 115 +/ 2 to 122 +/- 3 (6 months) and 124 +/- 2 (12 months) g/l, p = 0.03) despite a fall in EPO requirement (from 10,400 +/- 1400 to 8500 +/- 1300 (6 months) and 7600 +/- 1100 (12 months) U/week, p = 0.03). In contrast, CHD cohort had no change in EPO requirement (from 8300 +/- 1100 to 8100 +/- 1300 (6 months) and 8600 +/- 1000 (12 months) U/week, p > 0.05) or Hb concentrations (from 110 +/- 2 to 115 +/- 3 (6 months) and 115 +/- 2 (12 months), p > 0.05). There was a higher percentage of ESRD patients who did not require EPO in the NHD cohort (24% vs. 9.4%, p = 0.01). Lower Hb concentrations were noted in the CHD cohort despite higher iron saturation (0.25 +/- 0.01 (NHD) vs. 0.33 +/- 0.02 (CHD), p = 0.02) at the end of follow-up. CONCLUSIONS: Enhancing uremic clearance by NHD resulted in a rise in Hb and a fall in EPO requirement. PMID- 15786822 TI - Conversion of temporary hemodialysis catheters to tunneled hemodialysis catheters. AB - PURPOSE: To determine the feasibility and clinical outcomes of conversion of temporary to tunneled hemodialysis catheters using the same venous insertion site. METHODS: Data from 42 patients with existing temporary hemodialysis catheters referred for placement of tunneled hemodialysis catheters were retrospectively reviewed. In these patients, the temporary catheter was exchanged for a peel-away sheath, and a tunneled catheter was inserted using the existing venous access site. Technical success, procedural complications, and clinical outcomes were evaluated. Hemodialysis records were reviewed to assess catheter patency during a 30-day follow-up period. RESULTS: The study group consisted of 20 males and 22 females (mean age: 58 years). All 42 temporary catheters were successfully converted to tunneled hemodialysis catheters without immediate procedure-related complications. Follow-up data were available for 32 patients (total: 3038; median 71 catheter days). Nine catheters were removed for infection, yielding a catheter infection rate of 0.30/100 catheter days; three catheters were removed for blood flow <200 ml/min. 13 patients had catheters removed when catheters were no longer needed. Three patients died with working catheters. The patency rate was 72% at 30 days, with four catheters functioning at the end of the study period. CONCLUSION: Conversion of a temporary hemodialysis catheter to a tunneled hemodialysis catheter using the same venous insertion site is a safe procedure that avoids complications associated with venotomy and allows conservation of other central venous access sites. Patency and infection rates in these catheters are comparable to several studies of catheter exchange and de novo placement of tunneled hemodialysis catheters. PMID- 15786823 TI - Resolution of primary amyloidosis by melphalan and prednisolone: a case report. AB - We here report a case of a 50-year-old man who showed histologically evident resolution of primary amyloidosis by melphalan and prednisolone. The patient was admitted to our hospital for further evaluation of nephrotic syndrome and remarkable hepatomegaly with refractory ascites, on September 11, 1998. Laboratory tests at presentation showed nephrotic syndrome with slight renal impairment and elevation of the enzymes of the biliary system. Monoclonal light chains were not detected in the serum or urine by immunoelectrophoresis. A renal biopsy revealed global deposition of amyloid in all glomeruli, interstitium and blood vessels. Immunofluorescence staining was positive for kappa light chains. Liver biopsy specimens showed extensive deposition of amyloid along sinusoid walls. Bone marrow aspiration contained 7% plasma cells but no clusters or abnormal cells. Based on these findings, systemic AL- (amyloid light chain) amyloidosis was diagnosed, and the treatment with combinations of melphalan and prednisolone was started from October 1998 at intervals of 4-6 weeks. Renal impairment progressed, resulting in the initiation of maintenance hemodialysis in February 1999. Reinfusion of ascitic fluid into the hemodialysis circuit had been performed from March 1999 for refractory ascites, and ascites disappeared in July 1999. Furthermore, urinary output increased after 14 courses of chemotherapy. Renal function gradually ameliorated with a concomitant reduction in the enzymes of biliary system, and finally hemodialysis was discontinued in April 2001. Sixteen courses of chemotherapy were administered by April 2001. Proteinuria was negative in August 2001. A second renal biopsy was performed on November 20, 2001, which showed markedly decreased amyloid deposition and a proliferation of mesangial cells and increase in matrix in various degrees. We report a case of a patient with primary amyloidosis who was successfully treated by melphalan and prednisolone, resulting in marked resolution of renal amyloidosis. PMID- 15786824 TI - COX-2 inhibitor induced anuric renal failure in a previously healthy young woman. AB - Side effects of nonsteroidal anti-inflammatory drugs (NSAIDs) most commonly affect the gastrointestinal tract and the kidney. The recent release of selective cyclooxygenase-2 (COX-2) inhibitors has been associated with a decrease in adverse gastrointestinal effects. However, the nephrotoxic potential of these drugs still remains controversial. Here, we report the case of a previously healthy woman with reversible acute renal failure associated with eight days of anuria following the administration of valdecoxib, a newly released selective cyclooxygenase-2 inhibitor, during an episode of acute febrile pyelonephritis. We suggest that selective COX-2 inhibitors should not be used in patients with volume contraction and underlying renal disease. PMID- 15786825 TI - Ethylene glycol intoxication and xylitol infusion--metabolic steps of oxalate induced acute renal failure. AB - Acute renal failure is a major complication in patients with increased oxalate serum concentration. To describe the metabolic mechanisms of oxalate-induced glomerular and tubular damage, we report a case of ethylene glycol intoxication as well as a case of xylitol infusion in a patient with previously unknown primary hyperoxaluria type 1. Both patients presented with acute renal failure associated with histologically proven renal oxalate accumulation. This excessive oxalate overloading resulted from elimination and metabolization of ethylene glycol or xylitol. Thus, key enzymes in the elimination pathway of these substances represent targets for pharmacological treatment. Simultaneous hemodialysis is often necessary to reduce oxalate serum concentration. Whereas renal function of the ethylene glycol-poisoned patient recovered, the second patient who received xylitol infusion required chronic hemodialysis due to the unmasked hyperoxaluria type 1. Our cases demonstrate that patients with excessive endogenous oxalate generation are at high risk to develop acute renal failure. Therefore, to prevent end-stage renal failure in these patients, important clinical factors should be considered as indicators for the underlying cause: history of alcohol abuse and severe high anion gap acidosis for ethylene glycol intoxication or history of long-lasting parenteral nutrition for xylitol associated acute renal failure. PMID- 15786826 TI - Severe rhabdomyolysis and renal failure triggered by a sauna visit in sickle cell trait: a case report. AB - Sickle cell trait (SCT) is an usually asymptomatic hemoglobinopathy. Cases of sudden excertional deaths in individuals with SCT have been described. We here report an exceptional case of excessive rhabdomyolysis and acute renal failure triggered by a sauna visit in a 29 year-old African American with SCT. PMID- 15786827 TI - Superiority of standard interferon-alpha2b compared to pegylated interferon alpha2b (12 kDa) in a hemodialysis patient with chronic hepatitis C? AB - INTRODUCTION: Hepatitis C virus (HCV) infection represents an important problem for hemodialysis patients. Interferon-alpha (IFN-alpha) three times per week has been shown to clear HCV RNA in a substantial proportion of renal transplant candidates, and may thereby prevent the deleterious effect of immunosuppressive treatment on progression of liver disease in HCV-positive patients after renal transplantation. Data on the efficacy of the new pegylated interferons in hemodialysis patients are limited and general recommendations are absent. CASE: A 41-year-old Caucasian man infected with hepatitis C genotype 1b was admitted with a history of renal transplantation in 1990, and reintroduced hemodialysis in 1997 because of chronic rejection. Antiviral therapy with pegylated interferon-alpha2b (120 microg/oiw) and ribavirin (400 mg/tiw) was initiated. A virological and biochemical response with undetectable HCV-RNA was evident already after six weeks. Two weeks later, however, HCV-RNA became detectable again with 18.000 IU/ml. The treatment regimen was changed to standard-IFN-alpha2b (3 MU/tiw). Shortly thereafter, ribavirin had to be withdrawn because of severe anemia. After three weeks, hemoglobin level rebounded to values higher than 10 g/dl and a lower dose of ribavirin (200 mg/tiw) could be reintroduced. Virological and biochemical response occurred after switching to standard interferon-alpha2b within three months with good tolerance of antiviral combination treatment until the end of 48 weeks of therapy. The patient remained HCV-RNA-negative throughout follow-up of 36 weeks. ALT levels are still within normal limits and the patient is now waiting for a kidney transplantation. CONCLUSION: Considering the treatment course of this patient, IFN-alpha2b three times per week directly after hemodialysis seems to be superior to pegylated interferon-alpha2b once weekly in this case. The role of pegylated IFN-alpha2a for dialysis patients remains to be investigated. PMID- 15786828 TI - Thrombocytopenia and anaphylaxis secondary to heparin in a hemodialysis patient. AB - AIMS: We describe a rare case of anaphylaxis and thrombocytopenia whose cause was heparin used during hemodialysis sessions. CASE REPORT: A 77-year-old woman suffered five consecutive episodes of vomiting, tachypnea, wheezing or rales, immediately after initiating hemodialysis. In the first of these episodes, arterial pressure was undetectable. In all of the episodes there was evidence of the presence of hypoxia (always below 60 mmHg) and thrombocytopenia (always below l00,000/microl,, with partial platelets recovery among episodes. The episodes started immediately after hemodialysis sessions and heparin infusion; either sodium heparin or enoxaparin was used. Utilization of different filters was not able to stop the episodes. These were stopped when a switch from heparin to hirudin was tested. Tryptase levels, as a marker of mast cells activation and anaphylaxis, were not increased in two of the episodes which were assessed. IgG antibodies against heparin-PF4 complex was detected at high levels. DISCUSSION: A diagnosis of concomitant anaphylaxis and thrombocytopenia caused by sodium heparin and a low-molecular weight heparin (enoxaparin) were assumed. PMID- 15786829 TI - Recurrent peritonitis caused by Corynebacterium amycolatum in a patient undergoing continuous ambulatory peritoneal dialysis. PMID- 15786830 TI - Metastatic primitive neuroectodermal tumor presenting as acute renal failure and hypercalcemia. PMID- 15786831 TI - [Prophylaxis and treatment of postoperative purulent-septic complications in patients with perforative gastroduodenal ulcer]. AB - Experience of treatment of 43 patients, operated on for perforative gastroduodenal ulcer, was summarized. In 23 of them in the early postoperative period amoxyclav was applied for the treatment and prophylaxis, what have had assisted the reduction of the complications rate as well as the length of treatment of the patients in stationary. Good tolerance of the preparation was noted. PMID- 15786832 TI - [Autoimmune changes in patients with multiple gastroduodenal ulcers and their pathogenetic significance in the complex surgical treatment]. AB - There were examined 73 patients, suffering for multiple gastroduodenal ulcers and included in the main group, and 56 patients with solitary gastroduodenal ulcer, included in the comparison group. The lymphocytes migration impeding reaction (LMIR) was applied to characterize the autoimmune changes in both groups of patients. It was established, that in the patients with multiple gastroduodenal ulcers and with ulcers of small sizes the LMIR was essentially lowered in relation to all the autoantigenes investigated. The data obtained trust the important significance of the immunological investigation conduction in substantiation of the pathologic process pathogenesis, what must be taken into account during performance of surgical treatment and while applying the treatment rehabilitation measures after the operation. PMID- 15786833 TI - [Application of synthetic analogue of somatostatin--octrestatin in the treatment of gastroduodenal erosions and acute ulcers complicated by hemorrhage]. AB - Retrospective analysis of diagnostic and curative processes in Kyiv's city centre of delivering the aid in gastrointestinal hemorrhage for 1131 patients in 1999- 2003 yrs period was done. The classification of gastroduodenal erosions and an acute ulcers, complicated by hemorrhage, oriented on etiological factor involved, was proposed. Depending on etiological feature present, there were determined characteristic signs of mucosal affections, their localization and the hemorrhage severity. There was proved the expediency of introduction of the treatment results control, based on investigation of dehydrogenases activity, the intracellular glycogen content and the state of the lipids peroxidal oxidation. Clinical efficacy of synthetic analogue of somatostatin Octrestatin was studied in the treatment of gastroduodenal erosions and an acute ulcers in 26 patients. Application of Octrestatin have promoted the lipids metabolism normalization and acceleration of the mucosal ulcers healing by 2-3 days. PMID- 15786834 TI - [Reconstructive-restoration surgeries for nonspeciphic ulcerative colitis and colonic Crohn's disease]. AB - Results of surgical treatment of 88 patients were presented, to whom radical surgery for nonspeciphic ulcerative colitis and Krohn's disease was performed. In 92% of observations the first-stage operation consisted of subtotal colectomy procedure with formation of separate ileostoma and sygmostoma, which was modified in the clinic. The reconstructive-restoration operation was performed in 61 (69.4%) of patients in 12 months after the first operation. The necessity to estimate the degree of the excluded colon readiness for the reconstructive restoration operation performance was established, the terms of its application as well as the method and character of the operation were substantiated. The postoperative complications rate was (45,5+/-5,3)%. The result of reconstructive restoration operation was considered good in 12, fair--in 61, unfair--in 15 patients. All the patients are alive. PMID- 15786835 TI - [Method of the surgical treatment of nonspeciphic extrasphincteric rectal fistula]. AB - The method of treatment of nonspeciphic extrasphincteric rectal fistula was proposed, which consists of the fistula channel excision en bloc with external and internal (the anal sinus, which bears the fistula) apertures on perineum, subsequent layer by layer restoration of the rectal wall, as well as the sphincters and the anal levators muscles integrity (if necessary) and the formed cavity drainage. The operation was performed in 25 patients. Its performance have guaranteed the recurrences rate lowering as well as the postoperative incontinence occurrence. PMID- 15786836 TI - [Remote results of the treatment of proximal rectal cancer]. AB - There were presented the late results of surgical treatment in 425 patients, suffering upper ampullar and rectosygmoid rectal cancers, to whom transabdominal rectal resection with the duplicature anastomosis formation in 1989--2002 yrs was done. In 93.4% of observations, according to histological investigation data, adenocarcinoma was diagnosed. Concurrent diseases were noted in 226 (53.2%) of patients. The postoperative complications rate was 13.2%, postoperative mortality -2.6%. Late result of treatment was fair. PMID- 15786837 TI - [Quality of life in patients after laparoscopic and open interventions for colonic tumors]. AB - The analysis of results of surgical treatment of the colonic cancer patients, basing on the quality of life scale indexes, using questionnaire SF-36, was performed. There were compared the quality of life in patients, who have been operated, using laparoscopic (I group) and open (II group) interventions in 1998- 2003 yrs. The quality of life indexes in 7-8 days after the operation were higher in I group than in the II group. Positive influence of performance of laparoscopic intervention on quality of life trustworthy differed of such, achieved in the II group. In 3, 6 months, in 1 and 3 years after the operation the quality of life indexes were identical in both groups. PMID- 15786838 TI - [Functional state of the liver in patients with an acute destructive pancreatitis in the early postoperative period]. AB - The efficacy of conventional complex treatment and such, including pentoxyphylline, lipoic acid and the aminoacids mixture, was studied in early postoperative period in 64 patients with an acute destructive pancreatitis. The highest levels of general and direct bilirubins, the aminotransferases activity were established initially in those patients, in whom the polyorgan insufficiency syndrome (PIS) occurred subsequently. More significant normalization of the indexes under the influence of treatment was observed in patients without PIS, in patients with PIS the pharmacological resistance was registered. More favourable course of postoperative period was observed in patients, in whom pentoxyphylline, lipoic acid and the aminoacids mixture were included in complex of treatment. PMID- 15786839 TI - [Differentiated surgical tactic in patients with an acute biliary pancreatitis and the jaundice syndrome]. AB - The outcome was analyzed in 466 patients, treated for an acute pancreatitis in 1997--2003 yrs. In 187 (40.1%) of patients the jaundice had occurred. The surgical tactic was elaborated, depending on the jaundice origin. The extrahepatic biliary ducts pathology have had constituted the acute pancreatitis etiology in 119 (64%) patients and the alimentary causes--in 68 (36%). The predominant application of miniinvasive methods, such as endoscopic papillosphincterotomy with lithoextraction, laparoscopic cholecystectomy, transcutaneous puncture-draining interventions under ultrasonographic control, was based on the proved evidence of efficacy of the staged surgical treatment in patients with biliary acute pancreatitis and the jaundice syndrome; open method was used according to indications. PMID- 15786840 TI - [Efficacy of the surgical treatment for malignant heart tumors]. AB - Peculiarities of malignant cardiac tumors surgical treatment were studied, basing on analysis of 33 observations. Rhabdomyosarcoma was diagnosed in 9 patients, angiosarcoma--in 11, leyomyosarcoma--in 6, malignant mixoma--in 4, fibrosarcoma, chondrosarcoma and liposarcoma--each as a single observation. Special attention was paid to guarantee the maximal radicalism in the tumor excision, what made it necessary in 8 observations to perform additive surgical procedures, such as interatrial septum and atrial walls plasty, plastic operations on cardiac valves and the valves prostheses. The operation volume extension did not influence significantly its immediate result. PMID- 15786841 TI - [The tricuspid valve plasty in the surgical treatment of chronic pulmonary thromboembolism]. AB - Chronic pulmonary thromboembolism causes persistent rising of pressure in the pulmonary artery system. Durable existing of hypertension promotes the right ventricle dilatation together with tricuspid valve insufficiency occurrence, dramatizing the disease clinical course and prognosis. There were considered the surgical methods of the tricuspid cardiac valve insufficiency correction. The performance of the tricuspid valve plasty according to De Vega and using supporting ring of Carpentiere was proposed, basing on experience of surgical treatment of chronic pulmonary thromboembolism and postthrombotic pulmonary hypertension. PMID- 15786842 TI - [Anatomical correction of the main vessels transposition. Echocardiographic estimation of cardiac parameters preoperatively and in immediate period after the operation]. AB - Radical anatomic correction--operation of arterial switch--is performed in the main vessels transposition in majority of the world big cardiosurgical clinics, but its late results are studied insufficiently yet. Echocardiographic cardiac parameters preoperatively and in immediate postoperative period after anatomic correction in the main vessels transposition were presented. Comparative analysis of indexes, including estimation of anatomy and function of cardiac ventricles, depending on initial data and terms of the operation performance was conducted. PMID- 15786844 TI - [Decrease of the surgical risk level due to enhancement of the operative activity]. AB - The surgical risk dynamics was studied up, basing on the scientific investigations data, performed during last 30 years. Basing on controversies existed in majority of aspects while performing the risk estimation calculation the need for the process to gain objective was established. The mathematical modelling procedure was proposed for the risk estimation. There were studied the influence of the surgeon qualification on the operative treatment volume and the result, as well as the surgeons of highest qualification occupation when the quantity of severely ill patients in emergency surgery enhances. The results, witnessing the reduction of the surgical risk factors influence while operative activity enhances, were adduced. PMID- 15786843 TI - [Diagnostic and treatment aspects of cardiac and pericardial wounds]. AB - Basing on experience of treatment of 97 injured persons there was analyzed the course of cardiac and pericardial wounding, the conventional and modern diagnostic possibilities were compared, the staged approach, directed to the surgical treatment results improvement, was systematized. PMID- 15786845 TI - [Modern methods of the treatment of injured persons with combined vertebrospinal trauma]. AB - The issues of rehabilitational treatment of injured persons after polytrauma occurrence with compressive fracture of spine using electrostimulation were considered. The application of such an approach have had promoted the essential lowering of traumaticity and the treatment duration as well, the prognosis improvement, the length of temporary disability period reduction. PMID- 15786846 TI - [Role of nitric oxide and lipid peroxidation in pathogenesis of acute experimental pancreatitis]. AB - In experiment, conducted on 130 Wistar line rats, there was studied up the influence of nitric oxide on the peroxidal oxidation of lipids intensity, activity of enzymes of the antioxidant defense system and morphological changes in acute pancreatitis. Modern suggestions about the influence of the nitric oxide system on pathogenetic mechanisms of an acute pancreatitis and possibilities of correcting therapy were scrutinized. It was established, that the nitrates and the nitrites content in the blood serum of experimental animals correlates positively with the pancreatic oedema index and severity of its morphological changes as well. Intraperitoneal injection of 20% water solution of L-arginine in dosage 200 mg per 100 g of an animal body mass have had aggravated an acute pancreatitis course; intravenous injection of 1% water solution of methylene blue in dosage 1 mg per 100 g of an animal body mass caused pancreatoprotective effect. PMID- 15786847 TI - [Experience in the organ transplantations in the Institute of Surgery and Transplantology of AMS of Ukraine]. PMID- 15786848 TI - Cost effectiveness of thromboprophylaxis with a low-molecular-weight heparin versus unfractionated heparin in acutely ill medical inpatients. PMID- 15786849 TI - Asthma disease management: regression to the mean or better? PMID- 15786850 TI - Asthma disease management: regression to the mean or better? PMID- 15786851 TI - Achieving value in healthcare. PMID- 15786852 TI - The impact of Medicare modernization on coverage policy: recommendations for reform. PMID- 15786853 TI - Can an algorithm for appropriate prescribing predict adverse drug events? AB - OBJECTIVE: To evaluate whether a medication-appropriateness algorithm applied to pharmacy claims data can identify ambulatory patients at risk for experiencing adverse drug events (ADEs) from those medications. STUDY DESIGN: Cohort study. METHODS: We surveyed a random sample of 211 community-dwelling Medicare managed care enrollees over age 65 years who were identified by pharmacy claims as taking a potentially contraindicated medication (exposed enrollees) and a random sample of 195 enrollees who were identified as not taking such a medication (unexposed enrollees). The primary outcome of interest was the prevalence of self-reported events in previous 6 months. RESULTS: Ninety-nine (24.4% of total sample) respondents reported a total of 134 ADEs during the previous 6 months. Exposed enrollees had a significantly higher number of chronic conditions and were taking more prescription and nonprescription medications. However, the higher rate of self-reported ADEs among exposed enrollees was not statistically significant from that of unexposed enrollees (prevalence odds ratio = 1.42; 95% confidence interval [CI] = 0.90, 2.25). Only 1.5% (2/134) of the self-reported ADEs were attributed to a medication from the potentially contraindicated list. Instead, most ADEs were attributed to medications that are commonly used in older patients, including cardiovascular agents (21.6%), anti-inflammatory agents (12.2%), and cholesterol-lowering agents (7.9%). CONCLUSIONS: A medication appropriateness algorithm using pharmacy claims data was not able to identify a subgroup of enrollees at higher risk of experiencing an ADE from those medications. The vast majority of ADEs were attributable to commonly prescribed medications. PMID- 15786854 TI - Physician evaluations of care management practices in Medicaid programs. AB - OBJECTIVE: To assess differences in care management practices for Medicaid beneficiaries in predominantly commercial and predominantly Medicaid health plans. STUDY DESIGN: Physicians in the networks of 8 managed care plans participating in Medicaid programs were surveyed regarding the availability and usefulness of care management practices and the overall quality of care management. The responses of physicians in plans serving predominantly Medicaid enrollees were contrasted with the responses of physicians in predominantly commercial plans who cared for Medicaid enrollees. METHODS: Logistic regression analysis was used to calculate adjusted odds ratios relating to the availability of care management practices. Multiple regression techniques were used to construct comparisons of adjusted means relating to the usefulness of practices and the overall quality of care management. RESULTS: Physicians in predominantly commercial plans reported greater availability of care management practices. No patterns of differences were noted in ratings of the usefulness of practices if available. Physicians in predominantly commercial plans rated the quality of care management higher than physicians in predominantly Medicaid plans. However, there remains room for substantial improvement for commercial and other Medicaid contracting plans. CONCLUSIONS: Commercial plans add value to Medicaid programs, and efforts to discourage their withdrawal from participation are justified. However, physician evaluations support the potential for better care management in all types of contracting plans. PMID- 15786855 TI - Reduction in neonatal intensive care unit admission rates in a Medicaid managed care program. AB - BACKGROUND: Neonatal intensive care unit admission rates are an important birth outcome indicator for Medicaid managed care organizations. OBJECTIVES: To reduce neonatal intensive care unit admission rates by at least 15% and to maintain that reduction through implementation of a quality improvement program. STUDY DESIGN: The organization performed a longitudinal population-based review of its birth outcomes from 1997 through 2003, focusing on neonatal intensive care unit admission rates. The return-on-investment evaluation reflected attributable incremental program costs and resultant savings. METHODS: Interventions included enhanced identification and stratification of high-risk women with the use of a health risk assessment form; outreach through nursing care coordination offering home visits, transportation, support services, social work services, and connection with other community-based organizations; and implementation of a strong informatics structure. RESULTS: Neonatal intensive care unit admission rates decreased from 107.6 per 1000 births in 1998 to 56.7 per 1000 births in 2003. The return on investment from the incremental program enhancements was just over dollars 2 per dollars 1 expended. CONCLUSION: A program that identifies its high-risk pregnant enrollees in a timely fashion, provides outreach using a strong nursing care coordination and social work emphasis, and has an enhanced informatics structure can significantly affect birth outcomes for a Medicaid managed care population. PMID- 15786856 TI - Alternative strategies for Medicare payment of outpatient prescription drugs- Part B and beyond. AB - Reimbursement options for pharmaceuticals reimbursed under Medicare Part B (physician-dispensed drugs) are changing and the new comprehensive Part D Medicare outpatient drug benefit brings further changes. The Medicare Prescription Drug, Improvement and Modernization Act of 2003 (MMA) replaces traditional policy, of reimbursing Part B drugs at 95% of average wholesale price (AWP, a list price), with a percentage markup over the manufacturer's average selling price; in 2005 an indirect competitive procurement option will be introduced. In our view, although AWP-based reimbursement has been fraught with problems in the past, these could be fixed by constraining growth in AWP and periodically adjusting the discount off AWP. With these revisions, an AWP-based rule would preserve incentives for competitive discounting and deliver savings to Medicare. By contrast, basing Medicare reimbursement on a manufacturer's average selling price undermines incentives for discounting and, like any cost-based reimbursement rule, may result in higher prices to both public and private purchasers. Indirect competitive procurement for drugs alone, using specialty pharmacies, pharmacy benefit managers, or prescription drug plans, is unlikely to constrain costs to acceptable levels unless contractors retain flexibility to use standard benefit management tools. Folding Part B and Part D into comprehensive contracting with health plans for full health services is likely to offer the most efficient approach to managing the drug benefit. PMID- 15786857 TI - Variation in Medicare's local coverage policies: content analysis of local medical review policies. AB - OBJECTIVE: To assess variation in the content of Medicare's local medical review policies. STUDY DESIGN: Six case studies to compare differences in coverage policies by diagnosis codes, procedure codes, and indications for use. METHODS: All carrier policies from 48 carrier contracts (n = 5213) posted to the Centers for Medicare & Medicaid Services Web site were downloaded on May 31, 2001. All policies in the data set were coded based on a typology: new technology (NT), extensions of new technology (TE), and utilization management (UM) of widely used procedures. We identified policies addressing the same procedure or technology. We required at least 20 separate policies in each case study to allow meaningful comparisons. We randomly selected 1 case study of a diagnostic and 1 for a treatment modality from each policy type (NT, TE, and UM). RESULTS: Given previous research on local carriers, we expected to find variations among policies in each case study. We found substantial similarity, however, among policies covering the NT and TE types. We found significantly more variation among our UM-type case studies. CONCLUSIONS: Medicare legislation has called for greater coverage policy consistency in Medicare. This analysis on variation in policy content, part of a larger study on variation in Medicare's local coverage process, provides data on policy content differences. Policy reform should reflect the nature of and reasons for policy variation as suggested by the findings of this research. PMID- 15786858 TI - Nurses: many roles, one profession. PMID- 15786859 TI - Oklahoma works for better breast cancer screening and treatment. PMID- 15786860 TI - Why ovarian cancer awareness? PMID- 15786863 TI - Generational approaches to current nursing issues: how younger and older nurses can coexist. PMID- 15786864 TI - Value from involving professional staff nurses. PMID- 15786865 TI - Strategies for a successful retirement. PMID- 15786866 TI - Antibody recognition of synthetic peptides mimicking immunodominant regions of HIV-1 p24 and p17 proteins. AB - The gag gene of HIV-1 encodes a single open reading frame of 55 kDa that contains three subdomains: the matrix domain (p17), the capsid domain (p24) and the nucleocapsid domain (p15). The p24 and p17 proteins have a predominant alpha helical structure and perform important functions throughout the viral life cycle. The determination of gag-specific antibodies is important because declining titers of these antibodies herald clinical deterioration. In this work we present the results obtained on immunoreactiviy of synthetic peptides that mimic immunogenic alpha-helical regions of p24 and p17. The influence on the immunoreactivity of structural modifications in native sequences, including the addition of non immunogenic side chains: AAAC- and -CAAA on both side of minimal epitopes was evaluated in indirect and competitive enzyme immunoassays. The conformational characteristcs to the peptides were analysed by circular dichroism and these results were correlated with that obtained in the immunoassays. It was shown that the reactivity of peptides mimicking short alpha-helical regions of p24 and p17 is improved by adding short non immunogenic chains on both N- and C terminus. These modifications enhanced the immobilization of the peptides onto the solid support and allowed more accessibility to the minimal epitopes by specific antibodies, in solution. PMID- 15786867 TI - Virulence factors of non-O1 non-O139 Vibrio cholerae isolated in Cordoba, Argentina. AB - V. cholerae non-O1 non-O139 serogroups isolated from clinical and environmental sources in Cordoba, Argentina, were analyzed for the presence and expression of virulence genes. Most of the strains studied contained the genes toxR and hlyA, but lacked ctxA, zot, ace, tcpA and stn. The culture supernatants were tested for hemolytic and cytotoxic activity. The enterotoxic potential of the strains was studied in a rabbit ileal loop assay and their genetic profiles were compared by PFGE. The environmental strains varied in their virulence phenotype and showed no clonal relationships. The clinical strains were highly enterotoxic, hemolytic, proteolytic and showed indistinguishable PFGE profiles, although they differed in their cytotoxic activity. This is the first description, using cell culture and "in vivo" studies, of the virulence properties of non-O1 non-O139 V. cholerae from Argentina. PMID- 15786868 TI - [Analysis of a Yersinia enterocolitica isolated from human diarrheic feces in Argentina]. AB - Some serotypes of Yersinia enterocolitica might cause diarrheas and/or invasive infections. The aim of this work was to analyze virulence factors and associated markers in a strain of Y. enterocolitica isolated from human diarrheic feces. The strain analyzed was included in the biotype 1A. The virulence markers determination as well as the search of the genes vir F and ail, were negatives. However, it was demonstrated enterotoxin production at 20 degrees C, and at 37 degrees C in osmolarity conditions and pH similar to the human intestine. The enterotoxin presented reactivity for the infant mouse test, although it could not be proven the presence of yst gene by PCR. The results obtained by us, coincident with those of other investigators, indicated that certain clinical isolates of Y. enterocolitica of the biotype 1A ("avirulent"), could be the etiological agent of the illness through other mechanisms of virulence, that would differ from those previously characterized in species of enteropathogenic Yersinia. PMID- 15786869 TI - [Prevalence of mycobacteriosis and tuberculosis in a reference hospital, Cordoba province]. AB - Environmental mycobacteria (EM) constitute an important group of bacteria species found in the environment. They can colonize and occasionally produce disease in man. Sixteen thousand three hundred samples from 9300 adult symptomatic patients from the Hospital Regional of Tuberculosis in Cordoba were bacteriolocally investigated. The isolations were performed by culture on Lowenstein Jensen and Stonebrink culture media. The colonies of acid fast bacilli (AFB) were identified by biochemical and molecular tests. Among 716 culture positive cases, 684 (95.5%) were due to Mycobacterium tuberculosis complex and 32 to environmental mycobacteria. Serial samples allowed the confirmation of the etiological agent in culture and correlated with consistent clinical and radiological abnormalities. Seventy-five percent of these patients were affected by M. avium complex, 15.6% by M. fortuitum, 3.1% Mycobacterium kansasii and 6.3% Mycobacterium chelonae. Among tuberculosis cases, 94.5% and 5.5% had pulmonary and extrapulmonary disease respectively. PMID- 15786870 TI - [Immunoglobulins in patients with Nocardia brasiliensis actinomycetoma]. AB - Considering that some authors have reported an increasing of some immunoglobulins in actinomycetoma patients, in this study we propose to determine differential production of IgG1, IgG2, IgG3, IgG4 and IgGM in 25 patients with actinomycetoma and 25 healthy individuals from a mycetoma endemic area. Immunoglobulins were determined by ELISA technique. To sensibilize the plates, six Nocardia brasiliensis antigens were used: a crude antigen denominated NB and five derivatives (NB2, NB4, NB6, NB8 and NB10) obtained by their isoelectric point. Results showed that all IgG subclasses were higher in the patients' sera than in control sera, with a maximal difference to IgG3 and IgG4. To the latter subclass, six antigens were highly reactives. IgM levels were similar in both groups. As it occurs in other infections, in the actinomycetoma pathogenesis probably participate the increase or deficiency of a determined immunoglobulin class, as well as the relationship between different subclasses. PMID- 15786871 TI - [Listeria monocytogenes detection in different food products and environmental samples from a large chain of supermarkets in the city of Bahia Blanca (Argentina)]. AB - This work on Listeria monocytogenes detection in different foods was carried out between January 2002 and July 2003. Ninety cold-served cooked meats, sliced and packaged by different methods and 132 pieces of soft cheeses were studied. These products were analyzed using the presence/absence in 25 g criterion. L. monocytogenes was not found either in foods sliced over the counter or in controlled cheeses, but it was found in 10% of sliced cold-served foods and 5% of cut and cold-served meats vacuum packaged. These results led us to investigate the presence of these pathogen bacteria in different environmental samples. A hundred and fifteen points were swabbed including processing lines, raw materials, tools, and refrigerators. L. monocytogenes was found in 13.2% of the analyzed samples: 5% in packaging sector, 6.7% in meat processing lines and 1.5% in personalized sales. These results showed the presence of sites where the microorganism may reside and create reservoirs, so that routinary measures of hygiene and disinfection were increased. PMID- 15786872 TI - [Penicillium-inhibiting yeasts]. AB - The objective of this work was to establish the in vitro and in vivo inhibition of post-harvest pathogenic moulds by yeasts in order to make a biocontrol product. Post-harvest pathogenic moulds Penicillium digitatum, P. italicum, P. ulaiense, Phyllosticta sp., Galactomyces geotrichum and yeasts belonging to genera Brettanomyces, Candida, Cryptococcus, Kloeckera, Pichia, Rhodotorula were isolated from citrus fruits. Some yeasts strains were also isolated from other sources. The yeasts were identified by their macro and micro-morphology and physiological tests. The in vitro and in vivo activities against P. digitatum or P. ulaiense were different. Candida cantarellii and one strain of Pichia subpelliculosa produced a significant reduction of the lesion area caused by the pathogenic moulds P. digitatum and P. ulaiense, and could be used in a biocontrol product formulation. PMID- 15786873 TI - [Biological and enzymatic activities in salt-affected soils from Alto Valle de Rio Negro and Neuquen]. AB - Changes in the biological activity (number of bacteria g(-1) and CO2 production) and in the enzymatic activity (catalase, deshydrogenase, urease and phosphotriesterase) caused by the leaching of five soils affected by salts have been studied. The leaching decreased the electric conductivity (CE) and modified the type of dominant salts in the soils. Production of CO2 and the activity of the phosphotriesterase was significantly higher (p<0.05) in a leached soil (Torrifluventes Typical Centennial); the increment were 88% and 71%, respectively. The results showed that the decrease of the salinity by leaching did not produce significantly different results in most of the biotic parameters analysed. PMID- 15786874 TI - Effects of hydroclimatic anomalies on bacteriological quality of the Middle Parana River (Santa Fe, Argentina). AB - Lenitic and lotic aquatic environments were studied in the Middle Parana River basin, one of the largest rivers of the world (basin: 1,510,000 km2, length: 2,570 km). The Parana River (S 31 degrees 42' 04"; W 60 degrees 29' 39"), during El Nino-Southern Oscillation (ENSO) (1997-1998), registered maximum flows of 29,962 m3 s(-1) (more than twice the mean flow) and also positive pluvial anomalies (A) (from +120 mm to +161 mm). However, concentrations of Escherichia coli remained low, with values similar to the mean (p > 0.05) (1.3802 log10cfu/100 ml). Only a maximum peak (1.903 log10cfu/100 ml), significantly higher than the mean (p < 0.0001), was detected during the ascending phase of the flood pulse. During La Nina (September 1999-March 2000; Q = 11,255 m3 s(-1); A = 78 mm to -84 mm), concentrations of E. coli were significantly lower than the mean (p < 0.0001). During the post-Nina period, with normal flows (Q = 14,900 m3 s(-1)) but with strong positive pluvial anomalies (A = +118 mm to +377 mm), the fecal indicator density reached maximum values (up to 2.699 log10cfu/100 ml, p < 0.0001). The multiple regression model used with all the results obtained in the Parana River showed that variations in concentrations of E. coli are mainly explained by pluvial precipitations and hydrometric level (p = 0.001). Temporal patterns of concentrations of E. coli in a small, secondary course (Q = 101 m3 s( 1)) followed those of the Parana River, but concentrations and peaks of E. coli produced by exceptional rains showed a higher magnitude (maximum: 3.325 log10cfu/100 ml). In the floodplain lakes, away from urban centers, concentrations of E. coli were not associated to hydroclimatic anomalies, but directly to vegetable cover: maximum concentrations were always registered during periods that lakes remained covered by floating macrophytes (mainly Eichhornia crassipes); on the contrary, minimum concentrations were registered during periods without vegetable cover. PMID- 15786876 TI - Structure revisited. PMID- 15786875 TI - Help is on the horizon. PMID- 15786877 TI - PECSH-MNA makes contract gains. PMID- 15786878 TI - Concerted activity: protected or unprotected? PMID- 15786879 TI - MNA members testify before the Commission on Patient Safety. PMID- 15786880 TI - Looking to the future. PMID- 15786881 TI - Campaign promises: what's next for health care? PMID- 15786882 TI - 101 ways to improve nursing culture: communication. PMID- 15786883 TI - Why tuberculosis persists as a global problem. PMID- 15786884 TI - Optimal latent TB control methods. PMID- 15786885 TI - Can we get more HIV-positive tuberculosis patients on antiretroviral treatment in a rural district of Malawi? AB - The World Health Organization (WHO) has set a target of treating 3 million people with antiretroviral treatment (ART) by 2005. In sub-Saharan Africa, HIV-positive tuberculosis (TB) patients could significantly contribute to this target. ART (stavudine/lamivudine/nevirapine) was initiated in Thyolo district, Malawi, in April 2003, and all HIV-positive TB patients were considered eligible and offered ART. Despite this, only 44 (13%) of 352 TB patients were eventually started on ART by the end of November 2003. Most TB patients leave hospital after 2 weeks to complete the initial phase of anti-tuberculosis treatment (rifampicin-based) in the community, and ART is offered to HIV-positive TB patients after they have started the continuation phase of treatment (isoniazid/ ethambutol). ART is only offered at hospital, while the majority of TB patients take their continuation phase of anti-tuberculosis treatment from health centres. HIV-positive TB patients therefore find it difficult to access ART. In this paper, we discuss a series of options to increase the uptake of ART among HIV-positive TB patients. The main options are: 1) to hospitalise HIV-positive TB patients with a view to starting ART in the continuation phase in hospital; 2) to decentralise ART delivery so ART can be delivered at health centres; 3) to replace nevirapine with efavirenz so ART can be started earlier in the initial phase of anti-tuberculosis treatment. Decentralisation of ART from hospitals to health centres would greatly improve ART access. PMID- 15786886 TI - Highly active antiretroviral therapy (HAART) in adults with tuberculosis: current status. AB - The overlapping epidemiology of human immunodeficiency virus (HIV) infection and tuberculosis (TB) and the catastrophic consequences of the interactions between the two epidemics have led to increased morbidity and mortality due to HIV associated TB. While effective therapy is available for both conditions, there are major challenges in the concurrent treatment of HIV and TB co-infection. This review examines the interactions between HIV and TB infections and reviews the current status of highly active antiretroviral therapy (HAART) in patients with co-infection. Specific questions relating to optimal timing of concurrent HAART, challenges to concurrent HAART, optimal regimens and future considerations are discussed. PMID- 15786887 TI - WHO clinical staging of HIV infection and disease, tuberculosis and eligibility for antiretroviral treatment: relationship to CD4 lymphocyte counts. AB - SETTING: Thyolo district, Malawi. OBJECTIVES: To determine in HIV-positive individuals aged over 13 years CD4 lymphocyte counts in patients classified as WHO Clinical Stage III and IV and patients with active and previous tuberculosis (TB). DESIGN: Cross-sectional study. METHODS: CD4 lymphocyte counts were determined in all consecutive HIV-positive individuals presenting to the antiretroviral clinic in WHO Stage III and IV. RESULTS: A CD4 lymphocyte count of < or = 350 cells/microl was found in 413 (90%) of 457 individuals in WHO Stage III and IV, 96% of 77 individuals with active TB, 92% of 65 individuals with a history of pulmonary TB (PTB) in the last year, 91% of 89 individuals with a previous history of PTB beyond 1 year, 81% of 32 individuals with a previous history of extra-pulmonary TB, 93% of 107 individuals with active or past TB with another HIV-related disease and 89% of 158 individuals with active or past TB without another HIV-related disease. CONCLUSIONS: In our setting, nine of 10 HIV positive individuals presenting in WHO Stage III and IV and with active or previous TB have CD4 counts of < or = 350 cells/microl. It would thus be reasonable, in this or similar settings where CD4 counts are unavailable for clinical management, for all such patients to be considered eligible for antiretroviral therapy. PMID- 15786888 TI - Adherence to TB preventive therapy for HIV-positive patients in rural South Africa: implications for antiretroviral delivery in resource-poor settings? AB - OBJECTIVES: To explore patient and health worker perspectives on adherence to tuberculosis preventive therapy (TBPT), and to derive lessons for improving access to care amongst human immunodeficiency virus (HIV) infected individuals in resource-poor settings. DESIGN: Both quantitative and qualitative methods were employed. Patient records were reviewed for HIV-positive individuals attending a hospital-based HIV clinic between January 2000 and March 2002. Eighteen patients and two health care workers underwent in-depth interviews exploring perspectives around adherence. RESULTS: Of 229 HIV-positive clinic attendees, 94 (41.0%) were eligible for TBPT. Of 87 patients initiating a 6-month TBPT course of isoniazid 300 mg daily, 41 (47.1%) completed TBPT. Of the 46 interrupters, 16 (34.7%) did not return to the clinic after receiving their first dose of TBPT. Barriers to adherence included fear of stigmatization, lack of money for food and transport, the belief that HIV is incurable, competition between Western and traditional medicine, and a reluctance to take medication in the absence of symptoms. Disclosure of HIV status, social and family support, and a supportive clinic environment positively influenced adherence. CONCLUSION: Interventions to improve the accessibility and quality of the care delivery system have the potential to support adherence to TBPT and other HIV care regimens, including antiretroviral therapy. PMID- 15786889 TI - INH preventive therapy among adult HIV-infected patients in Thailand. AB - SETTING: Two HIV/AIDS clinics in Bangkok. OBJECTIVE: Although isoniazid (INH) preventive therapy (IPT) can reduce the risk of active TB among HIV-infected individuals, preventive therapy is rarely used in developing countries. The WHO recommends INH prophylaxis for tuberculin skin test (TST) positive HIV positives or for all HIV positives in countries with a high prevalence of latent TB if TST is unfeasible. It is not known whether IPT without TST will affect adherence. DESIGN: Prior to receiving IPT, 914 HIV-infected patients in Bangkok were randomized to TST or not. Adherence, measured by self-report and pill counts, and proportion completing therapy were evaluated. RESULTS: Adherence was 84.5% and 79.7%, by self-report, and 81.8% and 73.9% by pill count, respectively, in PPD positive and non-TST-screened subjects (adjusted OR 1.44, 95%CI 0.79-2.64 and 1.53, 95%CI 0.45-5.26). The drop-out rate before treatment was 6.3% in the TST screened and 1.7% in the non-TST screened subjects (OR 3.93, 95%CI 1.18-16.04). CONCLUSION: TST screening was not a predictor of adherence to IPT once therapy began, but it was associated with a higher drop-out rate prior to therapy. Acceptable levels of adherence were observed with both regimens. PMID- 15786890 TI - Is the combination of pyrazinamide plus rifampicin safe for treating latent tuberculosis infection in persons not infected by the human immunodeficiency virus? AB - SETTING: Nine public health care centres in four Spanish cities. OBJECTIVE: To evaluate the efficacy and safety of 2 months of rifampicin (R) plus pyrazinamide (Z) therapy (2RZ) compared with a 6-month course of isoniazid therapy (6H) for treating latent tuberculosis infection (LTBI). DESIGN: Multicentered, randomised, comparative and prospective trial conducted in HIV-seronegative contacts of infectious pulmonary TB cases. RESULTS: Of 352 individuals, 199 received 6H and 153 2RZ; 73% of contacts receiving 6H and 71% receiving 2RZ completed treatment (P = 0.73). Treatment interruption due to hepatotoxicity (ALT/AST > 5 times upper limit of normal) was observed in 10% of contacts in the 2RZ group and in 2.5% of the 6H group (P = 0.007). This higher than expected rate of hepatotoxicity in the 2RZ arm led to premature termination of the study. Severe or fatal liver injury was not detected. Liver function tests normalised after discontinuation of treatment. We conclude that the use of RZ should only be considered when other regimens are unsuitable and intensive monitoring of liver function is feasible. PMID- 15786891 TI - 'Kuyendela odwala TB'--visiting TB patients: the widening role of home-based care organisations in the management of tuberculosis patients in Lusaka, Zambia. AB - OBJECTIVE: To explore the widening role of home-based care (HBC) organisations in the management of TB patients in Lusaka, Zambia, in 1999. DESIGN: In a purposeful sample of eight HBC organisations and 1 hospice in Lusaka, 142 TB patients under HBC, 54 care givers, 42 TB patients not under HBC and 9 managers were interviewed. RESULTS: At least 50% of TB patients in Lusaka are cared for by HBC. The role of HBC in management of TB patients included food aid, practical and emotional support through the visits of voluntary care givers and, often, medical advice and treatment. TB diagnosis is carried out within the government health facilities. Five HBC organisations supplied anti-tuberculosis drugs, and three tried to carry out direct observation of treatment. The majority of the TB patients said their situation improved under HBC. Management was undermined by poor record keeping, sporadic anti-tuberculosis drug supplies, stigmatising preventive messages, limited supervision of care givers and poor coordination with the District Health Services. CONCLUSION: HBC organisations have become a key partner in TB control, looking after half the TB patients in Lusaka from diagnosis onwards, and complementing the public system. However, the quality of their management of TB and their partnership with government need to improve. PMID- 15786892 TI - Tuberculosis in Calgary, Canada, 1995-2002: site of disease and drug susceptibility. AB - SETTING: A centre managing all tuberculosis (TB) cases in the population of the Calgary region in Canada. OBJECTIVE: To facilitate the diagnosis and management of TB by examining the distribution of TB by site of disease and the prevalence and pattern of drug-resistant Mycobacterium tuberculosis in a north American setting. DESIGN: A retrospective analysis of data collected at the time of diagnosis of all TB cases recorded during the 8-year period 1995-2002. Data collected included demographic information, site of disease, method of diagnosis and M. tuberculosis drug susceptibility. RESULTS: During the 8-year period, 435 cases of TB were diagnosed: 49% had exclusively extra-pulmonary disease. Lymph node disease (usually cervical) accounted for 44% of the extra-pulmonary cases. A culture of M. tuberculosis resistant to at least one first-line anti-tuberculosis drug was detected in 16% of the initial isolates. All of the resistant strains were found in foreign-born patients and resistance was more common in patients from Asia (19%) and in previously treated patients (26%). CONCLUSION: Extra pulmonary TB is common in the foreign-born population in the Calgary region, and this pattern may be more widespread than is realised. Drug-resistant strains of TB were more common in foreign-born and previously treated patients. PMID- 15786893 TI - Sex-specific performance of routine TB diagnostic tests. AB - SETTING: City Council Chest Clinic, Nairobi, Kenya. OBJECTIVE: To determine to what extent the performance of smear microscopy is responsible for sex differences in notification rates. METHODOLOGY: Three sputum samples from TB suspects were subjected to smear microscopy with Ziehl-Neelsen (ZN) and auramine (FM) staining. Lowenstein-Jensen culture was used as the gold standard. RESULTS: Of 998 suspects, 600 (60%) were men and 398 (40%) women. The odds of detecting culture-positive patients with ZN was lower for women (OR 0.67). By examining the first spot specimen, ZN detected 35% of culture-positive males and 26% of culture positive females. These proportions increased to respectively 63% and 53% when examining three specimens, and to 79% and 74% when using FM. The sex difference reduced and became non-significant (P = 0.19) when adjusted for HIV; however, the numbers involved for HIV stratification were low. CONCLUSION: The performance of a diagnostic tool contributes to sex differences in notification rates and influences male/female ratios. Women were less likely to be diagnosed (P = 0.08), and when ZN was used they were less likely to be labelled as smear-positive TB (P < 0.01). The application of more sensitive diagnostic tools such as FM is to the advantage of women. PMID- 15786894 TI - Evaluation of new external quality assessment guidelines involving random blinded rechecking of acid-fast bacilli smears in a pilot project setting in Mexico. AB - SETTING: Laboratories in Mexico that support the national tuberculosis (TB) control program have been involved in an acid-fast bacilli (AFB) microscopy external quality assurance program which includes rechecking 100% of smears identified as AFB-positive by the local laboratories and 10% of smears identified as AFB-negative. Very few errors have been detected in Mexico using non-random selection and unblinded rechecking of the slides. OBJECTIVE: To evaluate the results from a 1-year pilot program involving blinded rechecking of randomly selected AFB slides from local TB laboratories in two Mexican states and determine its feasibility for future implementation. DESIGN: To reduce potential bias, laboratory staff from the National TB Laboratory, Institute for Epidemiological Diagnosis and Reference (InDRE), performed quarterly statistical sampling of AFB smears and on-site evaluations in local laboratories in each state. AFB smears were rechecked at the respective state laboratories with discordant results resolved at InDRE. RESULTS: A significantly greater percentage of errors was detected on the randomly selected, blinded AFB smears than on the non-randomly selected, unblinded smears. CONCLUSION: Random blinded rechecking provides more accurate estimates of AFB microscopy results, resulting in improved diagnosis and monitoring of treatment response. PMID- 15786896 TI - Tuberculin skin test: estimation of positive and negative predictive values from routine data. AB - OBJECTIVE: To estimate, using routinely available data, the predictive values of tuberculin testing with 2TU RT23 for detection of latent tuberculosis infection (LTBI) in the Dutch population as a basis for recommendations on cut-off values at various levels of infection prevalence. DESIGN: Smoothed distributions of TST reactions among 312 tuberculosis patients and 2848 healthy non-BCG-vaccinated persons were used to estimate the sensitivity and specificity at various cut-off values. RESULTS: Sensitivity was 98.9% at 5 mm, 95.4% at 10 mm and 79.8% at 15 mm cut-off. Specificity with the corresponding cut-offs was 95.3%, 96.3% and 97.1% before and 98.0%, 98.8% and 99.6% after adjustment for presumed LTBI. At 10 mm, the positive predictive value (PPV) was > 75% if the infection prevalence in the tested population was at least 10%, but strongly declined with lower prevalences. For lower prevalences a cut-off of 15 mm was proposed, as this results in a higher PPV without greatly affecting the negative predictive value. CONCLUSION: Estimation of the predictive values of the TST from routine data can be useful for establishing cut-off values for detection of LTBI in different populations with different tuberculin preparations. PMID- 15786895 TI - Application of lot sampling of sputum AFB smears for the assessment of microscopy centres. AB - SETTING: Designated microscopy centres (DMC) and additional microscopy centres (AMC) performing sputum acid-fast bacilli (AFB) microscopy, the District TB Centre (DTC) and a reference laboratory (RL). OBJECTIVES: To ascertain the feasibility of adopting lot sampling of AFB smears and to assess the performance of MCs employing Senior Tuberculosis Laboratory Supervisors (STLS) with no knowledge about the principles of quality assurance of AFB microscopy and RL based laboratory technicians with training on quality assurance for blinded checking of AFB smears. METHODS: Slides from MCs were transported to the DTC and the RL; 20 smears per month per MC were selected systematically; 1547 slides from DMCs and 726 from AMCs were checked, respectively, by STLSs at the DTC and by RL laboratory technicians. Discrepancies were resolved by referee. RESULTS: The discrepancy between MC laboratory technicians and STLSs at the DTC was 4.7%, compared to 1% at the RL. The STLSs and RL-based laboratory technicians had 70 and 2 errors, respectively. CONCLUSIONS: Lot sampling of AFB smears is feasible under field conditions. Assessment of MCs was more valid with RL-based technicians trained in principles of quality assurance of sputum AFB microscopy than with STLSs with no such training and working in the field. PMID- 15786897 TI - Low parathormone levels and hypercalcaemia in patients with pulmonary tuberculosis: relation to radiological extent of disease and tuberculin skin test. AB - AIM: We hypothesised that serum calcium (Ca) and parathormone (PTH) levels correlate with radiological extent and that there may also be a relationship between the tuberculin skin test (TST), serum Ca and PTH levels in patients with pulmonary tuberculosis (PTB). MATERIALS AND METHODS: Forty-four patients with active PTB and 33 healthy subjects were enrolled in the study. Serum Ca, PTH, magnesium and phosphate levels were measured in patients and controls and compared. Correlations were also investigated for TST values, erythrocyte sedimentation rate (ESR), the degree of radiological involvement, serum PTH and corrected Ca levels. RESULTS: There was a significant difference between the two groups for mean serum PTH and corrected Ca levels. Significant correlations were detected between radiological extent of disease and serum PTH levels, between TST values and serum PTH levels and between ESR and serum PTH levels. We suggest that abnormal Ca metabolism in PTB patients is related to the radiological extent of disease. Factors determining the radiological extent of disease, predominantly the patient's immune status, may have an important role in modulating Ca metabolism in PTB patients. PMID- 15786898 TI - Tuberculosis: what is the yield of associate investigations in non-BCG-immunized children with positive tuberculin skin tests? AB - SETTING: Urban county health department in Tarrant County, Texas, USA. OBJECTIVE: To determine the yield of associate investigations in non-bacille Calmette-Guerin (BCG) immunized children with positive tuberculin skin tests (TSTs). DESIGN: We compared the results of associate investigations of the contacts of 38 TST positive, non-BCG-immunized pre-school children with the results of contact investigations of 290 culture-confirmed persons with tuberculosis (TB). RESULTS: Associate investigations were more likely than contact investigations to identify persons with culture-confirmed TB and positive TSTs. Contacts identified through associate investigation of non-BCG-immunized pre-school children were 9.4 (95%CI 4.2-22.5) times more likely to have culture-confirmed TB and 2.3 (95%CI 2.0-2.7) times more likely to have positive TSTs than contacts of persons with culture confirmed TB. CONCLUSION: While conducting associate investigations is labor intensive, these data indicate that associate investigation of pre-school non-BCG immunized children should be assigned priority in TB control activities, at least equal to the importance of investigating the contacts of culture-proven TB. PMID- 15786899 TI - Development of a pyrosequencing approach for rapid screening of rifampin, isoniazid and ethambutol-resistant Mycobacterium tuberculosis. AB - SETTING: The need to minimize the transmission of drug-resistant Mycobacterium tuberculosis requires rapid identification procedures. OBJECTIVE: To develop a pyrosequencing approach for rapid screening of rifampin, isoniazid and ethambutol resistant M. tuberculosis based on characterization of resistance-associated hot mutations. DESIGN: Three pairs of PCR primers and three pyrosequencing sequencing primers for detecting mutations at codon 526 and 531 of the rpoB gene, codon 315 of the katG gene, and codon 306 of the embB gene were chosen. The sensitivity of the pyrosequencing approach was determined by assaying PCR products generated from 10-fold serial dilutions of the DNA from the H37Rv strain. The efficacy of the pyrosequencing approach was evaluated by analyzing clinical isolates with a known antibiotic phenotype. RESULTS: Resistance-associated hot mutations could be determined within 2 h after PCR amplification using pyrosequencing. About 45 fg DNA per reaction was required to obtain sufficient PCR products to produce a clear, accurate pyrosequencing pattern. No mutations were found in all 20 drug susceptible clinical isolates, while all isolates with mutations showed corresponding drug resistances. CONCLUSION: This pyrosequencing approach can be used for rapid screening of rifampin-, isoniazid- and ethambutol-resistant M. tuberculosis prior to standard drug susceptibility testing. PMID- 15786900 TI - Treatment of Mycobacterium bovis infected tuberculosis patients: San Diego County, California, United States, 1994-2003. AB - SETTING: A local tuberculosis control program. OBJECTIVE: To examine treatment completion rates of patients with Mycobacterium bovis disease and compare them with those of patients with M. tuberculosis disease. DESIGN: Retrospective review of data from a tuberculosis surveillance computer database. RESULTS: Data from 167 M. bovis patients and 928 M. tuberculosis patients were examined. Rates of treatment completion were not significantly different (78% vs. 82%, chi2 = 1.60, P = 0.174), although death was more frequent among M. bovis patients (15% vs. 7%). The median time to treatment completion was 94 days longer for M. bovis patients. CONCLUSION: Overall, treatment completion rates of M. bovis and M. tuberculosis patients were comparable, although the death rate was higher for M. bovis patients. The latter finding may be related to a particularly high death rate among HIV-infected M. bovis patients. Therapy duration was longer for M. bovis patients, probably because resistance to pyrazinamide prevented the use of a short-course (6-month) regimen. PMID- 15786901 TI - A potential role for nitric oxide pathway in tuberculous pleural effusion. AB - SETTING: Tuberculous pleural effusion leads to an immune response involving mainly immune and mesothelial cells. Nitric oxide (NO) produced by these cells may have antimycobacterial effects against Mycobacterium tuberculosis. OBJECTIVE: To investigate the possible role of NO in connection with the arginase enzyme, which controls the synthesis of NO through arginine depletion. DESIGN: Pleural fluid samples from 20 patients with tuberculous pleural effusion were used for arginase activity and NO level determination. Results were compared with those from 12 lung cancer, 12 pneumonia and 12 congestive heart failure (CHF) patients. RESULTS: Pleural arginase activity in tuberculosis patients was found to be significantly decreased compared to lung cancer and pneumonia groups, while the NO level was higher in tuberculosis patients. All groups except the CHF group had significant correlations between NO level and white blood cell count. Arginase activity and red blood cell count correlated significantly in lung cancer and CHF groups. CONCLUSION: The arginine-NO pathway seems to be involved in the pathogenesis of tuberculous pleural effusion. Decreased arginase activity may cause arginine accumulation, which may then lead to increased NO synthesis by immune and mesothelial cells, reflecting a host defence mechanism. PMID- 15786902 TI - Oral insertion of a flexible bronchoscope is associated with less discomfort than nasal insertion for Korean patients. AB - OBJECTIVE: The route of bronchoscope insertion varies between centres, without a firm rationale based on well-designed studies. We therefore compared nasal and oral insertion of a flexible bronchoscope and evaluated efficacy and patient satisfaction. DESIGN: Prospective randomised study of patients who underwent flexible bronchoscopy from May to September 2003 and who were randomly assigned to nasal and oral insertion approaches. RESULTS: Clinical characteristics, factors related to the procedure and patient satisfaction were analysed. In total, 307 patients were randomly assigned to the nasal (n = 158) or oral insertion groups (n = 149). No difference in baseline characteristics was identified between the groups. Insertion by the oral route was associated with a smaller amount of lidocaine use during the procedure (P = 0.04) and less frequent insertion site bleeding (P = 0.005). Patients assigned to oral insertion reported less discomfort during anaesthesia (P = 0.01) and scope insertion (P < 0.001), as well as less dyspnoea (P = 0.04) and coughing (P = 0.03). CONCLUSION: Oral insertion of a flexible bronchoscope was associated with less discomfort for patients than nasal insertion, although the route of insertion had no significant effect on outcome. PMID- 15786903 TI - The role of cotrimoxazole in HIV-coinfected TB patients: some concerns. PMID- 15786904 TI - [A good doctor can hardly be measured for quality]. PMID- 15786905 TI - [New strategy in type 2 diabetes tested in clinical trials. Glucagon-like peptide 1 (GLP-1) affects basic caused of the disease]. AB - A novel therapy for type 2 diabetes is based on the gut hormone, glucagon-like peptide-1 (GLP-1). GLP-1 is released from the gut during a meal intake and stimulates insulin secretion. The hormone also inhibits glucagon secretion, delays gastric emptying and induces satiety. It has been shown to reduce circulating glucose both under fasting conditions and after meal intake in subjects with type 2 diabetes. A problem in developing this novel therapy is that GLP-1 is rapidly inactivated by the enzyme dipeptidyl peptidase-4 (DPP-4), which results in a short half-life of the hormone requiring continuous infusion. Two strategies have been developed to circumvent this drawback. One strategy is the use of DPP-4 resistant GLP-1 receptor agonists (exenatide and liraglutide) and another strategy is to inhibit DPP-4 activity (LAF237). Both these strategies have been successful in clinical studies. PMID- 15786907 TI - [Fetal injury more and more common reason for second trimester abortion]. PMID- 15786906 TI - [Targeting--a new way to identify unknown tumor markers in blood vessels]. AB - The expression of specific molecules on the surface of vascular endothelial cells in tumours might be a key to anticancer therapy with angiostatic drugs. A new method to find these molecules on tumour vessels, targeting, is presented here. Some of these tumour-specific molecules have been identified by means of so called phage libraries. They are gene-manipulated phages, where the surface is decorated with randomly generated short peptides. After intravenous injection a few of the peptides, expressed on the surface of the phage, attach to complementary structures on the endothelial cell, as a ligand attaches to its receptor. Through biopsies and immunohistochemistry the phage can be isolated and identified. The part of the DNA of the phage that codes for the peptide-sequence of importance is sequenced. This seeking for such vessel-addresses can in the future be used for diagnostic purposes and also for local tumour-treatment. It is envisioned that cytotoxic drugs can be coupled to peptides on nanoparticles and act locally, in order to minimize toxic systemic side effects. PMID- 15786909 TI - [Anesthesia in hereditary peripheral muscular disease]. PMID- 15786908 TI - [Sudden cardiac death in athletes is usually caused by undiagnosed heart disease. Cardiac screening of young athletes under discussion]. AB - Sudden death during exercise in young athletes is usually caused by previously undiagnosed heart disease. The most frequent underlying diseases are hypertrophic cardiomyopathy, coronary artery anomalies and arrhythmogenic right ventricular cardiomyopathy. The possibility of carrying out screening to prevent these tragic cardiac deaths in young athletes has been discussed. A history of sudden cardiac death at a young age among relatives and/or the occurrence of exercise related symptoms in the active sportsperson may identify some of the individuals at risk. A pathological ECG is also a risk factor, especially in combination with a history of abnormal findings upon physical examination. For a correct evaluation of the young athlete, it is important to be aware of the normal variation of cardiac findings in athletes ("athlete's heart"). PMID- 15786911 TI - [Extensive public health work in necessary to achieve long-term effects on cardiovascular diseases]. PMID- 15786912 TI - [Intensive care must be well represented in the organ donation committee]. PMID- 15786910 TI - [Pfizer urged to respond to criticism and participate in the public debate]. PMID- 15786913 TI - [The use of beta blockaders in chronic heart failure shows how important the application of evidence based medicine really is]. PMID- 15786914 TI - [Public hearing about therapeutic alternatives in mental disease for knowledge dissemination]. PMID- 15786915 TI - [SSRI should be considered as the cause of hypoglycemias which are difficult to explain]. PMID- 15786916 TI - [Not everything worked perfectly in Thailand]. PMID- 15786917 TI - DEFRA sets out its framework for controlling bovine TB. PMID- 15786918 TI - Identification of Culicoides obsoletus (Diptera: Ceratopogonidae) as a vector of bluetongue virus in central Italy. AB - In 2001 and 2002, 235 outbreaks of bluetongue were observed in the Lazio and Tuscany regions of central Italy. During entomological surveillance Culicoides imicola, the main vector of bluetongue virus in the Mediterranean region, was detected in only 14 of 28 municipalities affected by outbreaks; Culicoides obsoletus was the most abundant species, contributing 83 per cent of individuals in catches, whereas C. imicola contributed only 2 per cent. In affected municipalities the maximum catch of C. obsoletus was 18,000 specimens, compared with 54 of C. imicola. In October 2002 bluetongue virus serotype 2 was isolated from a single pool of wild-caught, non-blood-engorged parous C. obsoletus inoculated on to BHK-21 cells. Its identity was confirmed by reverse transcriptase-PCR. PMID- 15786919 TI - Doppler sonography of the medial arterial blood supply to the coxofemoral joints of 36 medium to large breed dogs and its relationship with radiographic signs of joint disease. AB - The medial arterial supply to 68 of the 72 coxofemoral joints of 36 medium to large breed dogs was examined ultrasonographically. The medial circumflex femoral artery and three branches were identified; the artery and its transverse branch were identified in all 68 joints, and the deep branch was identified in 61 joints, and the ascending branch was identified in 63. However, the acetabular and obturator branches were not identified. The pulsatility index, the mean velocity and the peak systolic velocity of the medial circumflex femoral artery were determined and associated with a radiographic score of degenerative coxofemoral joint disease and a lath distraction index (LDI). In joints with a LDI greater than 0.35, the pulsatility index was significantly lower (P=0.023) and its mean velocity was higher (P=0.005). However, no significant associations were observed in individual dogs when the measurements in both joints were taken into account. PMID- 15786920 TI - Influence of outlying values and variations between sampling days on reference ranges for clinical chemistry in budgerigars (Melopsittacus undulatus). AB - Reference ranges for clinical chemistry variables were established for 99 budgerigars sampled monthly for 12 months. The data from the repeated measurements were compiled to obtain the most reliable population-based reference range defined to be relevant for the population of budgerigars over the whole year. Outlying values were excluded by reference to the population-based reference range and to previously determined values from the same individual, following the guidelines of the International Federation of Clinical Chemistry. The exclusion of outliers significantly reduced the calculated reference ranges. PMID- 15786921 TI - Diagnosis and incidence of Ornithobacterium rhinotracheale infections in commercial broiler chickens at slaughter. PMID- 15786922 TI - Pyogranulomatous meningoencephalitis in a goat due to Corynebacterium ulcerans. PMID- 15786923 TI - Clinical characterisation of natural scrapie in a native Spanish breed of sheep. PMID- 15786924 TI - Systemic rhabdomyonecrosis and acute tubular necrosis in a dog associated with wasp stings. PMID- 15786925 TI - Sarcocystis gracilis-like sarcocysts in a sheep. PMID- 15786926 TI - Australian tick paralysis in a dog imported into the UK. PMID- 15786927 TI - Wobbler syndrome in pedigree rams. PMID- 15786928 TI - Phenylbutazone and equine research. PMID- 15786929 TI - Pathogenic subspecies of Plasmodium relictum found in African birds. PMID- 15786930 TI - Suspect comparison. PMID- 15786931 TI - Secondary prophylaxis of venous thromboembolism. AB - Making decisions about any modality of secondary prophylaxis in patients with venous thromobembolism (VTE) has to balance the risk of bleeding induced by anticoagulants against the benefit of reducing the risk of recurrent disease. It has to be kept in mind that the magnitude of risk is not only defined by the number of events per time period but also by the impact of the event on the fate of the patient. With standard intensity Vitamin K antagonists, the risk of bleeding is more closely related to comorbidities than to other factors, e.g. age. The risk of VTE recurrence differs largely between patient groups. The criterion of presence, or absence of a permanent or transient clinical trigger factor for the actual VTE episode has a greater impact than an abnormal result in thrombophilia testing. The standard period ofsecondary prophylaxis for proximal deep vein thrombosis and for pulmonary embolism is three to six months. The concept of prolonging this period for several months according to the risk of recurrence is seriously challanged by the observation that the prolongation period seems to delay recurrencies rather than truly avoiding them. For this reason, patients who clearly are threatened by recurrent episodes should receive indefinitive secondary prophylaxis. This is the case for cancer patients, patients with the antiphospholipid syndrome, and those who belong to families with severe and symptomatic protein C, protein S, or antithrombin deficiencies. Patients with recurrent VTE, with idiopathic VTE, or with combined thrombophilic conditions may only benefit from indefinitive secondary prophylaxis if the bleeding risk of the anticoagulant regimen under consideration is very low. PMID- 15786932 TI - Effects of the Tibetan herbal preparation PADMA 28 on blood lipids and lipid oxidisability in subjects with mild hypercholesterolaemia. AB - BACKGROUND: Previous studies showed an anti-atherosclerotic effect of PADMA 28, an herbal formula based on Tibetan medicine. As the mechanisms of action are not fully understood, we investigated whether PADMA 28 may lower blood lipids and lipid oxidisability, and affect early endothelial dysfunction. PATIENTS AND METHODS: Sixty otherwise healthy subjects with total cholesterol > or = 5.2 mmol/l and < 8.0 mmol/l were randomly assigned to placebo or PADMA 28, 3 x 2 capsules daily, for 4 weeks (double-blind). Blood lipids (total, LDL-, and HDL cholesterol, triglycerides, Apo-lipoprotein A1 and B) and ex vivo lipid oxidisability were measured before and after treatment. In a subset of 24 subjects, endothelial function was assessed using venous occlusion plethysmography with intraarterial infusion of acetylcholine. Isolated LDL and plasma both untreated and pre-treated with PADMA 28 extract were oxidised by the radical generator AAPH. Conjugated diene formation was measured at 245 nm. RESULTS: Blood lipids did not change during the study in both groups. In contrast to previous reports in mild hypercholesterolaemia, no endothelial dysfunction was seen and, consequently, was not influenced by therapy. Ex vivo blood lipid oxidisability was significantly reduced with PADMA 28 (area undercurve: 5.29 +/- 1.62 to 4.99 +/- 1.46, p = 0.01), and remained unchanged in the placebo group (5.33 +/- 1.88 to 5.18 +/- 1.78, p > 0. 1). This effect persisted one week after cessation of medication. In vitro experiments confirmed the prevention of lipid peroxidation in the presence of PADMA 28 extracts. Persistent protection was also seen for LDL isolated from PADMA 28-pretreated blood after being subjected to rigorous purification. CONCLUSIONS: This study suggests that the inhibition of blood lipid oxidisability by PADMA 28 may play a role in its anti-atherosclerotic effect. PMID- 15786933 TI - The venous return simulator: an effective tool for investigating the effects of external compression on the venous hemodynamics--first results after thigh compression. AB - BACKGROUND: To present a virtual model, the venous return simulator (VRS), designed to compute venous hemodynamic variations when compression is applied to the leg. METHODS: The VRS defines a numerical network of the lower extremity and computes the dynamic variables (flow rate, venous diameter and internal pressure) for a defined external pressure. The VRS was based on physiological data from the literature and clinical studies on healthy subjects. Clinical correlations were required to confirm its validity; for this purpose, we carried out experiments simulating the conditions of a clinical trial, in which the diameter of superficial and deep veins was measured while increasing pressures (20, 40 and 60 mmHg.) were applied to the thighs of patients enduring deep valvular insufficiency and venous ulcers. The diameters and flow rates calculated using our VRS model were compared with the experimental data obtained at the same thigh compression levels. RESULTS: The numerical results of VRS are in good agreement with the clinical data obtained by Duplex, (R2 = 0.96). In accordance with the in vivo measurement the computed results show that only a pressure greater than 40 mmHg is able to reduce the venous diameter at thigh-level, both in the great saphenous vein and in the femoral vein. CONCLUSION: The venous return simulator computes lower limb hemodynamic parameters under static conditions. The good correlation existing between the VRS and the data obtained in a previous clinical study shows that this numerical approach could provide a useful means of predicting the hemodynamic consequences of compression therapy. PMID- 15786934 TI - Treatment of therapy-refractive ulcera cruris of various origins with autologous keratinocytes in fibrin sealant. AB - BACKGROUND: Evaluation of the effects of cultivated, subconfluent, autologous keratinocytes in fibrin sealant (BioSeed-S) on the healing of therapy-refractive chronic wounds. PATIENTS AND METHODS: Open observational study in 60 patients with chronic leg ulcers and impaired wound healing of various origins. After whole-skin excision and cultivation of the autologous keratinocytes, the suspended cells were applied to the preconditioned wound in fibrin sealant. Wound epithelization and wound size were recorded at defined times. RESULTS: Fifty-two of the 60 participating patients could be evaluated. After 6 weeks, 29 ulcers (55.8%) were healed. The mean epithelization increased between the 8th and 42nd postoperative day from 23% to 62.5%. In 50.0% of the patients, global assessment of the wound showed a high degree of epithelization or healing after 42 days. In 32.6% of treated patients, improvement was observed, while no healing tendency was to be found in 17.4%. CONCLUSION: The present observational study indicates that the transplantation of autologous keratinocytes suspended in fibrin sealant could be of advantage in the treatment of refractive leg ulcers. PMID- 15786935 TI - Development, validation and clinical use of the FLQA-I, a disease-specific quality of life questionnaire for patients with lymphedema. AB - BACKGROUND: It is known from clinical practice that lymphatic diseases can be associated with reductions of quality of life (QoL). Due to the lack of validated methods, only few studies however have systematically investigated the QoL in lymphedema. The aim of the study was 1) to develop a standardized QoL questionnaire specific for lymphedema and 2) to assess the QoL in these patients. PATIENTS AND METHODS: We developed and tested the FLQA-l, a novel QoL questionnaire developed specific for use in lymphedema on the basis of the previously validated FLQA vein questionnaire. The questionnaire consists of 92 items that refer to the following scales: Physical status, everyday life, social life, emotional well-being, treatment, satisfaction and profession/household. 392 patients with primary (n = 246) and secondary (n = 146) lymphedema were included in the validation study. RESULTS: The FLQA-l showed good internal consistency; Cronbach's alpha was higher than 0.75 in all scales. There were no floor and ceiling effects and satisfactory item selectivity. The test-retest reliability, sensitivity to change and convergent validity with other psychometric instruments were satisfactory. Clinically, patients with lymphedema showed markedly impaired QoL in all fields, compared to persons with early stage venous insufficiency, and comparable reductions of QoL, compared to patients with venous leg ulcer. CONCLUSION: These data indicate that the FLQA-l is a reliable and valid questionnaire for the assessment of QoL in lymphedema. Since the QoL is impaired in many patients with lymphedema, QoL evaluation may be helpful for clinical diagnostics as well as for outcome measurement of specific edema therapy. PMID- 15786936 TI - Duplex scanning criteria for selection of patients for internal carotid artery endarterectomy. AB - BACKGROUND: The value of carotid endarterectomy for stroke prevention depends on reliable identification of patients at higher risk for stroke from their internal carotid artery (ICA) occlusive disease than from surgery. This selection of patients is based on the degree of ICA stenosis. Therefore, preoperative diagnostic measures should strive for a prevalence independent probability for disease of 100%. Aim of this prospective study was to obtain clinically applicable duplex scanning criteria for ICA stenosis > or = 70% with a probability for disease of 100%. PATIENTS AND METHODS: In 124 ICA in 62 patients (79% male) angiography and duplex scanning were performed. Degree of stenosis was classified in 4 categories: I < 50%; II 50%-69%; III 70%-99%; IV 100%. Cohen's kappa statistic was used to estimate agreement between both methods within categories. To improve accuracy post-test likelihood for disease was calculated for each point on the receiver operating characteristics (ROC)-curve for peak systolic (PSV) and end-diastolic velocity (EDV), and cut-off points for velocity criteria were set at a positive likelihood of 100%. RESULTS: Diagnostic agreement was good with kappa = 0.77 (95% CI, 0.64-0.90; p < 0.001). For EDV a criterion of > or = 150 cm/sec was associated with a post-test likelihood for disease of 100%. For PSV no appropriate criterion could be detected. CONCLUSIONS: A probability of 100% for ICA stenosis > or = 70% can be achieved by mere preoperative duplex scanning. Vascular laboratory specific validation of duplex scanning criteria should consider prevalence independent post-test likelihood for disease to ensure the value of CEA for stroke prevention. PMID- 15786937 TI - Perioperative morbidity and mortality of carotid artery surgery under loco regional anaesthesia. AB - BACKGROUND: Loco-regional anaesthesia for carotid artery surgery has many advantages over general anaesthesia. It may be associated with a reduction in neurological, and equally important, non-neurological morbidity and mortality. However, sufficiently powered randomised controlled trials comparing general anaesthesia with local anaesthesia for carotid artery surgery are not yet published. Herein, we present our single centre experience of carotid endarterectomy under local anaesthesia and their respective procedure-related morbidity and mortality rates. PATIENTS AND METHODS: From January 1996 to December 2002, 1271 patients were operated on their carotid arteries. Of these, 1210 (95%) patients and 1355 carotid arteries were operated on in loco-regional anaesthesia and included in a prospective recording. The patients age ranged from 47 to 100 years (mean 70.5 years), 711 patients were male, 499 female. 496 patients (41%) were asymptomatic (Fontaine stage I), 460 have had a transient neurological deficit (TIA) prior to admission (Fontaine stage II) and 254 patients have had a stroke (Fontaine stage IV). RESULTS: The combined stroke rate was 2.2% (n = 30). The overall 30 day mortality was 0.2% (n = 3). The rate of haematoma indicating revision was 3% (n = 40). The revision in all cases was within 12 hours of surgery. No patient developed respiratory insufficiency after surgery. However, of the 40 patients with revision for haematoma, 4 (10%) needed prolonged respiratory assistance and one patient ultimately died of respiratory insufficiency and stroke. No cardiac mortality was observed. The over all rate of myocardial infarction observed postoperatively was 1.4% (n = 19), of which 1.1% (n = 15) were non q-wave infarcts. The combined shunting-rate for all stages was 18.6% (n = 252). CONCLUSION: Morbidity and mortality of carotid endarterectomy in loco-regional anaesthesia is comparable to recently published single-centre results. Patients with severe COPD, usually unsuitable candidates for general anaesthesia, can also be treated safely. PMID- 15786938 TI - Distinct factors correlating with adverse cardiac events after major vascular surgery. AB - INTRODUCTION: Cardiovascular complications remain the principal cause of both morbidity and mortality after major vascular surgery. The well-known coincidence between vascular disease and coronary artery disease provided the rationale for a detailed analysis of major perioperative cardiovascular complications in their relation to preoperative and intraoperative parameter METHODS AND PATIENTS: 90 patients scheduled to undergo either femoral-popliteal bypass (n = 74) or repair of an infrarenal aortic aneurysm (n = 16) were prospectively included in the study. All patients had no signs of unstable cardiac disease and required no cardiac testing. Both preoperative and intraoperative parameter were correlated to adverse cardiac events (cardiac death and myocardial infarction -MI). RESULTS: Univariate analysis identified the following parameter to be significantly related to cardiac complications: prior MI and intraoperative hypertension (systolic blood pressure above 200 mmHg). In contrast perioperative betablocker therapy was revealed to be protective. In multivariate analysis the history of MI and intraoperative hypertension correlated with poor cardiac outcome. CONCLUSIONS: Our results underline the importance of the individual history in predicting perioperative risk and corroborate the beneficial effects of long standing beta-blocker therapy. Additionally the significance of stable intraoperative hemodynamic parameter is demonstrated. PMID- 15786939 TI - [Spontaneous dissection of the subclavian artery and its branches]. AB - A 51-year-old woman suffering from vertigo presented to our cerebrovascular laboratory. MRI revealed a right-sided cerebellar infarction. On ultrasound examination, we found a dissection with wall hematoma of the right subclavian artery extending to the first segment of the vertebral artery and to the thyrocervical trunc, as well as a dissection of the left vertebral artery in the cranio-cervical junction. On follow-up evaluation, all stenoses resulting from dissection were recanalized within 2 months and findings of wall hematoma disappeared completely within 7 months. There was no evidence of a traumatic or iatrogenic cause of dissection; an aortic dissection was excluded by MRI. So we diagnosed a spontaneous dissection of the subclavian artery--an extremely rare but apparently existing vascular condition. PMID- 15786940 TI - Acroparesthesia and acral arterial occlusions as first manifestations of essential thrombocythemia. AB - Thrombocytosis is either caused by a reactive process (secondary thrombocytosis) or by a clonal bone marrow disorder The latter category includes essential thrombocythemia with bleedings and thrombotic complications as major causes of illness and death in this patients. We describe a 43-year-old man with a 6 months history of acroparesthesia in his toes. Half a year after onset of these symptoms, he noticed a bluish discoloration of digit V of his left foot. On first presentation physical examination revealed a bluish discoloration of all toes and a cold and blue digit V of the left foot. Peripheral pulses were all palpable, normal ankle systolic pressure measurements and normal pulse volume recordings except for digit V of the left foot were found. Laboratory tests revealed thrombocytosis of 800000/microliter. On treatment with acetylsalicylacid, prostanoids intravenously and low molecular weight heparin, the patient became asymptomatic and pulse volume recording of digit V was normalized. After exclusion of cardial or vascular sources of embolism by utrasonography bone marrow aspirate and biopsy supported the diagnosis of essential thrombocythemia. PMID- 15786941 TI - Perineal bruising: a sinister sign for leaking iliac aneurysms? AB - Ruptured false iliac aneurysms carry high mortality of up to 57%. It should be suspected whenever a patient who had previous aortic aneurysm repaired presents with bizarre lower abdominal symptoms. In the literature, different presentations have been reported, e.g progressive hydronephrosis, small intestinal obstruction, chronic venous insufficiency, arterio-venous fistula, and gross haematuria from arterio-vesical fistula. We report a patient suffering from a ruptured iliac aneurysm associated with a new sign that has not been described hitherto: perineal bruising. PMID- 15786942 TI - Endotension--a late complication successfully treated by open repair. AB - Endotension is a late complication following endovascular treatment of abdominal aortic aneurysm (AAA). A 83-year-old male had a successful endovascular repair of a 5.6 cm diameter AAA. During the follow-up period it was marked that the aneurysm continued to increase in size, became 6.6 cm maximal diameter and pulsatile with no evidence of endoleak. On laparotomy no endoleak was identified and the graft was left in situ. Postoperatively and on follow-up the patient remains asymptomatic while the aneurysm continued to appear shrunk. PMID- 15786943 TI - Coil embolization of an infected superior gluteal artery pseudoaneurysm caused by methicillin-resistant Staphylococcus aureus. AB - We describe the successful selective coil embolization of an infected superior gluteal pseudoaneurysm secondary to methicillin-resistant Staphylococcus aureus (MRSA) in a 36-year old women. The patient presented with a long history of drug abuse and perisacral abscesses due to chronic sacroilitis. The chosen strategy provides a safe and successful management of infected false gluteal artery aneurysm. PMID- 15786944 TI - Case report, "Endovascular treatment of subclavian pseudoaneurysm causing delayed brachial plexus paralysis", F. Schellhammer und H.-J. Muller, VASA 2004; 33: 239 241. PMID- 15786945 TI - Crunch time. PMID- 15786946 TI - PACS is a crowd-pleaser in healthcare. PMID- 15786948 TI - Stitching up surgical cost. PMID- 15786949 TI - Making beautiful music together. PMID- 15786947 TI - Lab link to patient safety. PMID- 15786950 TI - Safeguarding data integrity from power failure. PMID- 15786951 TI - Four F's equal A+. PMID- 15786952 TI - Quality with teeth. PMID- 15786953 TI - Healthcare IT tipping point? PMID- 15786955 TI - [Effect of some imidazole-4,5-dicarboxylic acid derivatives on the activity of N methyl-D-aspartate (NMDA) receptors]. AB - The results of experiments on mice showed that some imidazole-4,5-dicarboxylic acid derivatives injected into lateral cerebral ventricles produce a dose dependent convulsant or anticonvulsant effects, that is, possess the properties of partial NMDA receptor agonists. The most promising partial NMDA receptor agonist selected for further investigation is 2-propylimidazole-4,5-dicarboxylic acid. PMID- 15786954 TI - [Dilept: a tripeptoid neurotensin analog combining neuroleptic activity with positive mnemotropic action]. AB - The effect of N-caproyl-L-prolyl-L-tyrosine methyl ester (GZR-123, Dilept), a new antipsychotic drug with a dipeptide structure, on cognitive functions was studied using the passive avoidance conditioned reflex (PACR) test. It was found that the drug improved both the acquisition and retrieval of PACR under the conditions of undertraining and prevented the amnesic effect of transcorneal electroshock. The neurotransmitter analysis of these effects using the corresponding blockers showed the ability of dilept to facilitate the central glutamatergic (NMDA type) and cholinergic (of both muscarine and nicotine types) neurotransmission. No evidence of the involvement of the GABAergic system was observed. The experimental data on the combination of antipsychotic effect with memory improving activity suggests that dilept may be effective in cases of negative symptoms of schizophrenia and psychotic manifestations in Alzheimer disease. PMID- 15786956 TI - [Peculiarities of the temporal organization of the behavioral response to caffeine in rats]. AB - Chronic administration of a standard dose of caffeine (40 mg/kg) facilitated the formation of more clear circadian mobility rhythm in rats. The test animals belonging to various chronotypes exhibited a significant difference in sensitivity with respect to this psychostimulant. Rats possessing a more pronounced rhythm organization showed a greater increase in the rhythm amplitude, with the acrophase more significantly shifted toward early night hours. It is suggested that the individual chronotype features have to be taken into account in experimental and clinical pharmacology. PMID- 15786957 TI - [The rhythm-modulant effect of lithium hydroxybutyrate on the background of electrolytic lesion of the right suprachiasmatic nucleus of hypothalamus]. AB - Experimental lesion of the right suprachiasmatic nucleus of hypothalamus in rats facilitates manifestations of the free-running locomotor rhythm in winter solstice, but decreases the entrainment of the light cycle of this rhythm in the period of summer solstice. The administration of lithium hydroxybutyrate in both illumination regimes compensates the influence of lesion of the right suprachiasmatic nucleus. Features of the observed effect depend on the circadian phase of drug administration. PMID- 15786958 TI - [Stress-limiting properties of a new tetrahydropyrrolopyrazine derivative]. AB - The effect of compound PV-174, selected from a series of tetrahydropyrrolopyrazine derivatives was studied on a model of immobilization stress in rats. Pretreatment with PV-174 decreases the stress-induced involution of thymus and spleen, hypertrophy of cpinephrosis, and gastric ulceration. The drug also blocks the stress-enhanced lipid peroxidation in lungs and myocardium, while not influencing the effect of kenalog (an exogenous glucocorticoid). It is suggested that one of the possible mechanisms of the antistressor effect of PV 174 is related to the drug action upon CNS. PMID- 15786959 TI - [Gabaergic mechanism of cerebrovascular and neuroprotective effects of afobazole and picamilon]. AB - The role of GABAergic mechanisms in realization of the effects of afobazole and picamilon on the cerebral circulation was studied in rats. It is found that the cerebrovascular effect of afobazole significantly decreases on the background of the GABA receptor antagonists bicuculline and picrotoxin. This is evidence of an important role of the GABA system in the its cerebrovascular and neuroprotective activity of afobazole. The results of experiments with picamilon showed that the cerebrovascular effect of this drug is not affected by receptor blocking with bicucullin. However, the blocking of chloride channels of the GABA receptors by picrotoxin significantly decreased the effect of picamilon on the cerebral blood flow. PMID- 15786960 TI - [The antiarrhythmic effect of (-)-U-50,488 in rats with acute ischemia and reperfusion of heart is mediated by kappa1-opioid receptor activation]. AB - Pretreatment with a selective kappa1 opioid receptor (OR) agonist (-)-U-50,488 (1 mg/kg, i.v.) prevented the development of arrhythmias induced by occlusion (10 min) and reperfusion (10 min) in ketamine anesthetized rats, while the treatment with a less active enantiomer (+)-U-50,488 in the same dose produced no such effects. Preliminary intravenous administration of a selective kappa1 OR antagonist norbinaltorphimine (9 mg/kg) fully abolished the antiarrhythmic effect of (-)-U-50,488, while the kappa2 OR antagonist quadazocine (3 mg/kg) did not eliminate this effect. The injections of norbinaltorphimine or quadazocine alone did not influence the incidence of model arrhythmias caused by the occlusion and reperfusion. It was concluded that kappa1 OR stimulation favors an increase in cardiac tolerance to the arrhythmogenic action of occlusion and reperfusion. PMID- 15786961 TI - [Effect of cardiocyclide on the electrophysiological parameters of heart in experiments with vagus nerve excitation in narcotized dogs]. AB - The new class III antiarrhythmic agent cardiocyclide effectively suppresses atrioventricular fibrillations caused by the vagus nerve excitation in narcotized dogs. The electrophysiological effect of cardiocyclide was studied by method of programmed electric stimulation of myocardium on the background of excitation of the a peripheral segment of the right vagus by current pulses of increasing frequency. On this background, cardiocyclide exhibited characteristic effects manifested primarily in the repolarization and the AV node conduction. In addition, the drug eliminated suppression of the sinus and atrioventricular nodes and changes in the atrioventricular conduction caused by the vagus nerve. This drug effect was dose-dependent. The action of cardiocyclide in response to activation of the parasympathetic nervous system was independent of the frequency of the induced heart rate. PMID- 15786962 TI - [The role of alpha2-adrenergic and I1-imidazoline receptors in the effects of clonidine and moxonidine on isolated large intestine of mice]. AB - The ability of clonidine and moxonidine to interact with alpha2-adreno- and I1 imidazoline receptors was studied on isolated segments of large intestine of mice. Both drugs induced dose-dependent contractions in longitudinal muscles of the intestine segments. In both cases, the drug action was almost equally decreased by pretreatment with of yohimbine (alpha2-adrenoreceptor agonist with low affinity to I1-imidazoline receptors) and efaroxan (I1-imidazoline receptor agonist with low affinity to alpha2-adrenoreceptors). Analysis of the ratios of the antagonist activities (pA2) of yohimbine and efaroxan with respect to clonidine and moxonidine, as well as the relative selectivity of the two antagonists suggested that the action of both drugs on the large intestine is realized predominantly via alpha2-adrenoreceptors. PMID- 15786963 TI - [Gestagenic and contraceptive activity of new synthetic progesterone analogs in experimental animals]. AB - The gestagenic activity of new 17alpha-hydroxyprogesterone analogs was studied in experiments on infantile female rabbits (Clauberg-McPhail assay) and ovariectomized animals (Comer-Allen assay). The new steroidal preparations produced significant secretory transformations in immature rabbit endometrium. The maximum gestagenic activity was observed for butagest, while the minimum effect was produced by duhydrogesterone (dufaston). Butagest, megestrol capronate, and AMOL isopropyl ester showed the ability to maintain pregnancy in ovariectomized female rabbits. The compositions of gestagens (0.8 mg/kg) with ethinylestradiol (0.04 mg/kg) produced high contraceptive effect in rats. PMID- 15786964 TI - [Activation and blocking of 5-HT1A receptors influence on the immune response in CBA mice]. AB - 5-HT1A type serotonin receptors influence the immunomodulating action of the selective preparations 8-OH-DPAT (5-HT1A receptor agonist) and WAY-100635 (5-HT1A receptor antagonist) in CBA mice. The activation of 5HT1A receptors with 8-OH DPAT (1 mg/kg) decreased, while their blocking with WAY-100635 (1 mg/kg) increased the reaction intensity at the peak of response to immunization with ram erythrocytes. Preliminary blocking of the 5-HT1A receptors with WAY-100635 prevented the inhibiting action of 8-OH-DPAT. PMID- 15786965 TI - [Effect of melatonin on the glucocorticoid-induced immunity suppression studied in vitro]. AB - The effect of epiphyseal hormone melatonin on the cortisole-induced decrease in peripheral blood mononuclear cell (PBMC) proliferation and changes in T-helper type 1 cytokine (IFN-gamma) and type 2 cytokine (IL-4) expression was studied in vitro in cell culture supernatants. Cortisol (10(-6) M) decreased PBMC proliferation, suppressed IFN-gamma production, and did not influence IL-4 production. Melatonin in physiological concentrations (10(-11) M) partly reduced the cortisol-induced PBMC proliferation. This effect was not associated with changes in the IFN-gamma and IL-4 production in the cell culture. PMID- 15786966 TI - [Phagocyte activity of umbilical cord blood neutrophils in vitro in the presence of leucine]. AB - Leucine produces dose-dependent effect on the phagocyte activity of umbilical cord blood neutrophils in vitro. In an optimum dose of 0.1 mg/ml, leucine favors the completion of phagocytosis and increases the phagocyte index and phagocyte number. PMID- 15786967 TI - [Effect of ascorbic acid on the renal transport of endogenous nitrates and nitrites in humans]. AB - Effect of ascorbic acid on the renal excretion of endogenous nitrates and nitrites under aqueous-salt load was studied in practically healthy females. The administration of ascorbic acid in a dose of 10 mg/kg body weight under these conditions favored a decrease in the renal excretion of endogenous nitrates due to modulation of their renal clearance mechanism. PMID- 15786968 TI - [Effect of ethanol and 4-methylpyrazole on the immunotoxicity of ethylene glycol]. AB - The results of experiments on Wistar rats showed that acute poisoning with 1.0 LD50 of ethylene glycol decreased the nonspecific resistance of the organism (manifested by increased lethality caused by experimental infection, decreased average lethal dose of E. coli, and decreased mean effective lifetime of experimental animals) and reduced the antibody formation to predominantly a thymus-dependent antigen, natural and antibody-dependent cell cytotoxicity, and delayed type hypersensitivity reaction. 4-Methylpyrazole and (to a greater extent) ethanol potentiated the immunotoxic action of ethylene glycol. PMID- 15786969 TI - [Studying the efficacy of a series of quinoxaline and azolo[a(b)]quinoxaline derivatives as P2X receptor antagonists]. AB - The influence of a series of 14 quinoxaline and azoloquinoxaline derivatives on the P2X and P2Y receptor mediated response was studied in vitro on isolated rat and guinea pig tissues. Most of the compounds studied did not affect the response, while paratolylhydrazone-3-benzoyl-1,2-dihydro-2-oxoquinoxaline exhibited antagonism with respect to the P2X receptor mediated response, while not influencing the P2Y mediated relaxation. It is suggested that this compound can be used for the synthesis of new effective P2 receptor antagonists. PMID- 15786970 TI - [Effects of triamcinolone acetonide on leukocyte distribution in blood system, mononuclear liver infiltration, and immune response under conditions of stress induced hypoxia in rats]. AB - The effect of triamcinolone acetonide (2 mg/kg) on the distribution of morphologically mature leukocytes in the blood system, the mononuclear liver infiltration, and the immune response was studied in rats under conditions of stress-induced hypoxia. Administered under these conditions, the drug produces a less pronounced effect on the content of circulating lymphocytes, the lymphoid cell number in the bone marrow, and the number of hepatocytes and monocytes/macrophages in the liver. However, the post-stressor immunodepressant effect of triamcinolone acetonide was increased and accompanied by the development of hepatic damage. PMID- 15786971 TI - [Antihypoxant activity in a series of 2-aminothiazole homologs]. AB - The antihypoxic activity of a series of 2-aminothiazole homologs and the related N-syccinamic acids was studied on four hypoxia models. All 2-aminothiazoles possess antihypoxant properties, which increase with the complexity of the molecular structure. The attachment of a succinic acid fragment at the amino group reduced toxicity, but also decreased the antihypoxant activity of drugs. PMID- 15786972 TI - [Studying the diffusion of piyavit through rabbit eye cornea]. AB - The results of experiments on rabbits showed that the new drug piyavit, possessing pronounced thrombolytic properties, readily diffuses through cornea to the anterior chamber of eye. It is suggested that the biologically active components of piyavot facilitate the drug penetration through the corneal barrier. This favors maximum drug concentration and positively influences the therapy of traumas and eye diseases accompanied by hemorrhages. PMID- 15786974 TI - Governor Granholm announces Michigan Opportunity Partnership. PMID- 15786973 TI - [Paracetamol: advantages and drawbacks in the treatment of dystrophic destructive lesions in connective tissues]. AB - The advantages and drawbacks of using paracetamol in the treatment of dystrophic and destructive lesions in connective tissues are considered. PMID- 15786975 TI - What we all want, what we all need, what we all can do. PMID- 15786976 TI - 30 years of collective bargaining result in autonomy, voice in practice for UMPNC. PMID- 15786977 TI - E&GW leadership summit frames the future of MNA. PMID- 15786978 TI - Patients need the voice of nursing to ensure safety and quality. PMID- 15786979 TI - Political involvement builds safe patient care initiatives. PMID- 15786980 TI - Why MNA for the representing agent? PMID- 15786981 TI - 101 ways to improve nursing culture: respect. PMID- 15786982 TI - Nursing congresses benefit YOU. PMID- 15786983 TI - Mining the canine genome. Identification of genes helps breeders and researchers. PMID- 15786984 TI - Views dental study as flawed. PMID- 15786985 TI - Internships abound in coastal cities. PMID- 15786986 TI - What is your diagnosis? Panosteitis. PMID- 15786987 TI - Anesthesia case of the month. Hyperkalemia. PMID- 15786988 TI - Theriogenology question of the month. Endometrial cups. PMID- 15786990 TI - Antidotes in food animal practice. PMID- 15786989 TI - Revenues, expenses, and returns on resources for private veterinary practices in the United States, 2001 and 2003. PMID- 15786991 TI - Evaluation of aerobic bacteriologic culture of epidermal collarette specimens in dogs with superficial pyoderma. AB - OBJECTIVE: To evaluate a method of aerobic bacteriologic culture of epidermal collarette specimens from dogs with superficial pyoderma and compare results with those for aerobic bacteriologic culture of abdominal skin specimens in healthy dogs. DESIGN: Prospective study. ANIMALS: 22 dogs with epidermal collarettes and 24 healthy dogs. PROCEDURE: Dry sterile cotton swabs were rolled across epidermal collarettes or hairless areas of abdominal skin in healthy dogs and submitted for aerobic bacteriologic culture. Hemolytic colonies of gram-positive-staining cocci were tested for catalase production, and if results were positive, a coagulase test was performed. Colonies with coagulase activity were tested for the ability to ferment mannitol. Antimicrobial susceptibility testing was performed on all Staphylococcus spp that were isolated. RESULTS: S. intermedius was isolated from collarettes in 18 of 22 dogs with superficial pyoderma but not from healthy dogs. Estimated sensitivity and specificity of the culture method were 81.8% and 100%, respectively. There were no significant differences in the ability to culture S. intermedius, the number of S. intermedius isolates without resistance to antimicrobials, and the number of S. intermedius isolates resistant to penicillin G when comparing dogs with superficial pyoderma for the first time and dogs with recurrent pyoderma, dogs that did or did not receive concurrent antimicrobials, and dogs with and without underlying allergic disease. CONCLUSIONS AND CLINICAL RELEVANCE: Bacteriologic culture of epidermal collarette specimens was a simple and reliable method for identification of S. intermedius in dogs with superficial pyoderma, regardless of history of pyoderma or current antimicrobial use. PMID- 15786992 TI - Incidence of and risk factors for adverse events associated with distemper and rabies vaccine administration in ferrets. AB - OBJECTIVE: To determine incidence of and risk factors for adverse events associated with distemper and rabies vaccine administration in ferrets. DESIGN: Retrospective cohort study. ANIMALS: 3,587 ferrets that received a rabies or distemper vaccine between January 1, 2002, and December 31, 2003. PROCEDURES: Electronic medical records were searched for possible vaccine-associated adverse events. Adverse events were classified by attending veterinarians as nonspecific vaccine reactions, allergic reactions, or anaphylaxis. Patient information that was collected included age, weight, sex, cumulative number of distemper and rabies vaccinations received, clinical signs, and treatment. The association between potential risk factors and occurrence of an adverse event was estimated with logistic regression. RESULTS: 30 adverse events were recorded. The adverse event incidence rates for administration of rabies vaccine alone, distemper vaccine alone, and rabies and distemper vaccines together were 0.51%, 1.00%, and 0.85%, respectively. These rates were not significantly different. All adverse events occurred immediately following vaccine administration and most commonly consisted of vomiting and diarrhea (52%) or vomiting alone (31%). Age, sex, and body weight were not significantly associated with occurrence of adverse events, but adverse event incidence rate increased as the cumulative number of distemper or rabies vaccinations received increased. In multivariate logistic regression analysis, only the cumulative number of distemper vaccinations received was significantly associated with the occurrence of an adverse event. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that in ferrets, the risk of vaccine associated adverse events was primarily associated with an increase in the number of distemper vaccinations. PMID- 15786993 TI - Analgesic efficacy of preoperative administration of meloxicam or butorphanol in onychectomized cats. AB - OBJECTIVE: To determine analgesic efficacy and adverse effects of preemptive administration of meloxicam or butorphanol in cats undergoing onychectomy or onychectomy and neutering. DESIGN: Randomized controlled study. ANIMALS: 64 female and 74 male cats that were 4 to 192 months old and weighed 1.09 to 705 kg (2.4 to 15.5 lb). PROCEDURE: Cats received meloxicam (0.3 mg/kg [0.14 mg/lb], s.c.) or butorphanol (0.4 mg/kg [0.18 mg/lb], s.c.) 15 minutes after premedication and prior to anesthesia. A single blinded observer measured physiologic variables, assigned analgesia and lameness scores, and withdrew blood samples for each cat at baseline and throughout the 24 hours after surgery. Rescue analgesia (butorphanol, 0.4 mg/kg, i.v. or s.c.) or administration of acepromazine (0.025 to 0.05 mg/kg [0.011 to 0.023 mg/lb], i.v.) was allowed. RESULTS: Meloxicam-treated cats were less lame and had lower pain scores. Cortisol concentration was higher at extubation and lower at 1, 5, and 12 hours in the meloxicam-treated cats. Fewer meloxicam-treated cats required rescue analgesia at 3, 5, 12, and 24 hours after extubation. General impression scores were excellent or good in 75% of meloxicam-treated cats and 44% of butorphanol treated cats. There was no treatment effect on buccal bleeding time; PCV and BUN concentration decreased in both groups, and glucose concentration decreased in meloxicam-treated cats. CONCLUSIONS AND CLINICAL RELEVANCE: Preoperative administration of meloxicam improved analgesia for 24 hours without clinically relevant adverse effects in cats that underwent onychectomy or onychectomy and neutering and provided safe, extended analgesia, compared with butorphanol. PMID- 15786994 TI - Red blood cell transfusions in cats: 126 cases (1999). AB - OBJECTIVE: To determine the number of and reasons for RBC transfusions, incidence of acute transfusion reactions, prevalence of blood types, volume of blood administered, change in PCV, and clinical outcome in cats. DESIGN: Retrospective study. ANIMALS: 126 cats that received RBC transfusions. PROCEDURE: Medical records of cats that received whole blood or packed RBC transfusions were reviewed for signalment, blood type, pre- and post-transfusion PCV, volume of blood product administered, clinical diagnosis and cause of anemia, clinical signs of acute transfusion reactions, and clinical outcome. RESULTS: Mean volume of whole blood administered i.v. was 172 mL/kg (7.8 mL/lb) versus 9.3 mL/kg (4.2 mL/lb) for packed RBCs. Ninety-four percent of cats had blood type A. Mean increase in PCV among all cats was 6%. Fifty-two percent of cats had anemia attributed to blood loss, 10% had anemia attributed to hemolysis, and 38% had anemia attributed to erythropoietic failure. Acute transfusion reactions occurred in 11 cats. Sixty percent of cats survived until discharge. CONCLUSIONS AND CLINICAL RELEVANCE: RBC transfusions resulted in an increase in PCV in cats with all causes of anemia in this study. The rate of death was greater than in cats that did not receive transfusions, but seriousness of the underlying disease in the 2 groups may not be comparable. Death rate of cats that received transfusions was not attributable to a high rate of transfusion reactions. Results confirm that pretransfusion blood typing or crossmatching is required to minimize the risk of adverse reactions. PMID- 15786995 TI - Indications for and outcome of positive-pressure ventilation in cats: 53 cases (1993-2002). AB - OBJECTIVE: To determine indications for and outcomes of positive-pressure ventilation (PPV) in cats, document ventilator management, and identify factors associated with outcome. DESIGN: Retrospective study. ANIMALS: 53 cats that underwent PPV. PROCEDURE: Information on signalment, history, concurrent diseases, clinical findings, results of venous blood gas analyses and clinicopathologic testing, treatment, ventilator settings, and outcome was retrieved from the medical records. Data for cats that survived were compared with data for cats that died or were euthanatized while undergoing PPV RESULTS: PPV was initiated for management of respiratory failure (36 cats [68%]), cardiac arrest (9 [17%]), neurologic impairment (6 [11%]), and nonresponsive hypotension (2 [4%]). Eight cats (15%) survived, 19 (36%) died, and 26 (49%) were euthanatized while undergoing PPV. Cats that survived had a longer duration of ventilation than did those that died or were euthanatized and had a significantly higher incidence of ventilator-associated pneumonia. Signalment and ventilator settings were not associated with outcome. Cats that had no clinical evidence of pulmonary disease but required PPV because of primary neurologic disease had a higher survival rate (2/6) than did cats that required PPV because of respiratory failure (5/36), cardiac arrest (1/9), or nonresponsive hypotension (0/2). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the survival rate for cats requiring PPV may be lower than reported survival rates for dogs. Death was attributable to progressive respiratory failure, non-responsive hypotension, kidney failure, or neurologic impairment. PMID- 15786996 TI - Clinical, clinicopathologic, radiographic, and ultrasonographic abnormalities in cats with ureteral calculi: 163 cases (1984-2002). AB - OBJECTIVE: To determine clinical, clinicopathologic, radiographic, and ultrasonographic abnormalities in cats with ureteral calculi. DESIGN: Retrospective study. ANIMALS: 163 client-owned cats. PROCEDURE: Medical records were reviewed, and information on signalment, history, clinical signs, and results of clinicopathologic testing and diagnostic imaging was obtained. RESULTS: The number of cats in which ureterolithiasis was diagnosed each year increased progressively during the study period. Clinical signs tended to be non specific and included inappetence, vomiting, lethargy, and weight loss. A combination of survey radiography and abdominal ultrasonography revealed ureteral calculi in 66 of 73 (90%) cats in which the diagnosis was confirmed at surgery or necropsy. Ultrasonography revealed that ureteral calculi were causing ureteral obstruction in 143 of 155 (92%) cats. One hundred thirty-four of 162 (83%) cats had azotemia, 84 of 156 (54%) had hyperphosphatemia, and 22 of 152 (14%) had hypercalcemia. Urinary tract infection was documented in 10 of 119 (8%). Fifty eight of 76 (76%) cats with unilateral ureterolithiasis had azotemia and 33 (43%) had hyperphosphatemia, indicating impairment of renal function in the contralateral kidney or prerenal azotemia. Ultrasonographic imaging of the contralateral kidney in cats with unilateral ureteral calculi suggested that preexisting renal parenchymal disease was common in cats with ureterolithiasis. Ninety-one of 93 (98%) ureteral calculi contained calcium oxalate. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that abdominal imaging should be performed in all cats with chronic nonspecific signs or with acute or chronic renal failure to rule out ureterolithiasis. Preexisting renal disease may be common in cats with ureteral calculi. PMID- 15786997 TI - Management and outcome of cats with ureteral calculi: 153 cases (1984-2002). AB - OBJECTIVE: To determine outcome of medical and surgical treatment in cats with ureteral calculi. DESIGN: Retrospective study. ANIMALS: 153 cats. PROCEDURE: Medical records were reviewed. Owners and referring veterinarians were contacted for follow-up information. RESULTS: All cats were initially treated medically before a decision was made to perform surgery. Medical treatment included parenteral administration of fluids and diuretics to promote urine production and passage of the ureteral calculus and supportive treatment for renal failure. Ureteral calculi in the proximal portion of the ureter were typically removed by ureterotomy, whereas ureteral calculi in the distal portion of the ureter were more likely to be removed by partial ureterectomy and ureteroneocystostomy. Ureterotomy could be performed without placement of a nephrostomy tube for postoperative urine diversion. Postoperative complication rate and perioperative mortality rate were 31% and 18%, respectively. The most common postoperative complications were urine leakage and persistent ureteral obstruction after surgery. Chronic renal failure was common at the time of diagnosis and continued after treatment, with serum creatinine concentration remaining greater than the upper reference limit in approximately half the cats. Twelve-month survival rates after medical and surgical treatment were 66% and 91%, respectively, with a number of cats dying of causes related to urinary tract disorders, including ureteral calculus recurrence and worsening of chronic renal failure. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that medical and surgical management of ureteral calculi in cats are associated with high morbidity and mortality rates. Treatment can stabilize renal function, although many surviving cats will continue to have impaired renal function. PMID- 15786998 TI - Surgical and nonsurgical management of sagittal slab fractures of the third carpal bone in racehorses: 32 cases (1991-2001). AB - OBJECTIVE: To compare results (ie, return to racing and earnings per race start) of surgical versus nonsurgical management of sagittal slab fractures of the third carpal bone in racehorses. DESIGN: Retrospective study. ANIMALS: 32 racehorses (19 Thoroughbreds, 11 Standardbreds, and 2 Arabians). PROCEDURE: Medical records and radiographs were reviewed to obtain information regarding signalment and treatment. Follow-up information was obtained from race records. Robust regression analysis was performed to evaluate earnings per start in horses that raced at least once before and after injury. RESULTS: 22 (69%) horses raced at least once after treatment of the fracture. All 7 horses treated by means of interfragmentary compression raced after treatment, and horses that underwent interfragmentary compression had significantly higher earnings per start after the injury than did horses treated without surgery. Eight of 9 horses treated by means of arthroscopic debridement of the damaged cartilage and bone raced after treatment, but only 7 of 16 horses treated without surgery (ie, stall rest) were able to return to racing after treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that racehorses with sagittal slab fractures of the third carpal bone have a favorable prognosis for return to racing after treatment. Horses treated surgically were more likely to race after treatment than were horses treated without surgery. PMID- 15786999 TI - Evaluation of serum enzyme activities as predictors of passive transfer status in lambs. AB - OBJECTIVE: To determine the associations between serum IgG concentration and serum activities of gamma-glutamyltransferase (GGT), alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and pseudocholinesterase for the potential use of these serum enzymes as predictors of passive transfer status in neonatal lambs. DESIGN: Prospective observational study. ANIMALS: 47 Sardinian lambs from birth to 2 days old. PROCEDURE: Serum enzyme activities were measured by use of commercially available kits and a clinical biochemical analyzer. Serum IgG concentration was determined by single radial immunodiffusion. Associations between serum IgG concentration and the activity of each serum enzyme were established by use of regression analysis. RESULTS: A significant correlation was detected between serum IgG concentration and serum GGT activity in 1- and 2-day-old lambs. Minimal correlations were detected between serum IgG concentration and serum alkaline phosphatase activity in 1-day-old lambs and serum pseudocholinesterase activity in 1- and 2-day-old lambs. No significant associations were detected between serum IgG concentration and serum activities of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase. A multiple linear regression model was accurate for the estimation of the natural logarithm of serum IgG concentration as a function of the natural logarithm of serum GGT activity and of the age of lambs at the time of sampling (adjusted R2 = 0.89). This model was then used to calculate the serum GGT activity equivalent to various serum IgG concentrations for 1- and 2-day-old lambs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that passive transfer status in neonatal lambs can be successfully predicted by measurement of serum GGT activity but not by measurement of the other enzymes tested. PMID- 15787000 TI - Evaluation of patterns of removal and associations among culling because of lameness and sow productivity traits in swine breeding herds. AB - OBJECTIVE: To characterize patterns of removal and evaluate the associations among culling because of lameness and sow productivity traits among culled gilts and sows. DESIGN: Cross-sectional study. SAMPLE POPULATION: Data from a convenience sample of 11 farms pertaining to the removal of 51,795 gilts and sows from January 1991 to December 2002. Mean culling and mortality (death and euthanasia) rates for all inventoried gilts and sows ranged from 23% to 50% and 4.7% to 9.5%, respectively. PROCEDURE: An analysis of categories of removal (cull, death, or euthanasia) and reasons for removal of gilts and sows was performed. Multivariate logistic regression was used to determine associations among culling because of lameness and sow productivity traits among culled gilts and sows. RESULTS: Among sows that were removed, the proportion of parity > or = 1 sows that died (both death and euthanasia) was > 3 times the proportion of parity > or = 1 sows that were culled within 20 days after farrowing. Among lame sows that were removed, the proportion of parity > or = 1 sows that died (death and euthanasia) was higher than the proportion of parity > or = 1 sows that were culled within 20 days after farrowing. Among sows that were removed, the proportion of sows that died (deaths and euthanasia) was higher during lactation than nonlactation. This was also observed among lame sows that were removed. CONCLUSIONS AND CLINICAL RELEVANCE: The proportion of death among removed sows, especially lame sows, was higher during lactation than nonlactation. Results indicated that risk of death is not the same for sows throughout their lifetime. PMID- 15787001 TI - mobilePDR: a mobile medical information system featuring update via Internet. AB - This paper presents an analysis of usability of mobile prescription reference systems in medical practice, and presents implementation of mobilePDR (Physician's Desk Reference). Various aspects of mobilePDR are discussed: main functions, information content, performance and design issues, independent evaluation with other similar tools, and lessons learned from the system development. PMID- 15787002 TI - A direct-sequence spread-spectrum communication system for integrated sensor microsystems. AB - Some of the most important challenges in health-care technologies have been identified to be development of noninvasive systems and miniaturization. In developing the core technologies, progress is required in pushing the limits of miniaturization, minimizing the costs and power consumption of microsystems components, developing mobile/wireless communication infrastructures and computing technologies that are reliable. The implementation of such miniaturized systems has become feasible by the advent of system-on-chip technology, which enables us to integrate most of the components of a system on to a single chip. One of the most important tasks in such a system is to convey information reliably on a multiple-access-based environment. When considering the design of telecommunication system for such a network, the receiver is the key performance critical block. The paper describes the application environment, the choice of the communication protocol, the implementation of the transmitter and receiver circuitry, and research work carried out on studying the impact of input data characteristics and internal data path complexity on area and power performance of the receiver. We provide results using a test data recorded from a pH sensor. The results demonstrate satisfying functionality, area, and power constraints even when a degree of programmability is incorporated in the system. PMID- 15787003 TI - Performance analysis of multiplexed medical data transmission for mobile emergency care over the UMTS channel. AB - In this paper, a third-generation universal mobile telecommunications system (UMTS) solution for the delivery of biomedical information from an ambulance to a hospital is presented. The joint transmission of voice, real-time video, electrocardiogram signals, and medical scans in a realistic cellular multiuser simulation environment is considered, taking into account the advantages and particularities of UMTS technology for such transmission. The accomplishment of quality of service constraints for different services is investigated and quantitative results are provided in order to demonstrate the feasibility of using UMTS technology for emergency care services on high-speed moving ambulance vehicles. PMID- 15787004 TI - Real-time classification of ECGs on a PDA. AB - The new advances in sensor technology, personal digital assistants (PDAs), and wireless communications favor the development of a new type of monitoring system that can provide patients with assistance anywhere and at any time. Of particular interest are the monitoring systems designed for people that suffer from heart arrhythmias, due to the increasing number of people with cardiovascular diseases. PDAs can play a very important role in these kinds of systems because they are portable devices that can execute more and more complex tasks. The main questions answered in this paper are whether PDAs can perform a complete electrocardiogram beat and rhythm classifier, if the classifier has a good accuracy, and if they can do it in real time. In order to answer these questions, in this paper, we show the steps that we have followed to build the algorithm that classifies beats and rhythms, and the obtained results, which show a competitive accuracy. Moreover, we also show the feasibility of incorporating the built algorithm into the PDA. PMID- 15787005 TI - Bluetooth telemedicine processor for multichannel biomedical signal transmission via mobile cellular networks. AB - One of the emerging issues in m-Health is how best to exploit the mobile communications technologies that are now almost globally available. The challenge is to produce a system to transmit a patient's biomedical signals directly to a hospital for monitoring or diagnosis, using an unmodified mobile telephone. The paper focuses on the design of a processor, which samples signals from sensors on the patient. It then transmits digital data over a Bluetooth link to a mobile telephone that uses the General Packet Radio Service. The modular design adopted is intended to provide a "future-proofed" system, whose functionality may be upgraded by modifying the software. PMID- 15787006 TI - Mobile tele-echography: user interface design. AB - Ultrasound imaging allows the evaluation of the degree of emergency of a patient. However, in some instances, a well-trained sonographer is unavailable to perform such echography. To cope with this issue, the Mobile Tele-Echography Using an Ultralight Robot (OTELO) project aims to develop a fully integrated end-to-end mobile tele-echography system using an ultralight remote-controlled robot for population groups that are not served locally by medical experts. This paper focuses on the user interface of the OTELO system, consisting of the following parts: an ultrasound video transmission system providing real-time images of the scanned area, an audio/video conference to communicate with the paramedical assistant and with the patient, and a virtual-reality environment, providing visual and haptic feedback to the expert, while capturing the expert's hand movements. These movements are reproduced by the robot at the patient site while holding the ultrasound probe against the patient skin. In addition, the user interface includes an image processing facility for enhancing the received images and the possibility to include them into a database. PMID- 15787007 TI - A tele-operated mobile ultrasound scanner using a light-weight robot. AB - This paper presents a new tele-operated robotic chain for real-time ultrasound image acquisition and medical diagnosis. This system has been developed in the frame of the Mobile Tele-Echography Using an Ultralight Robot European Project. A light-weight six degrees-of-freedom serial robot, with a remote center of motion, has been specially designed for this application. It holds and moves a real probe on a distant patient according to the expert gesture and permits an image acquisition using a standard ultrasound device. The combination of mechanical structure choice for the robot and dedicated control law, particularly nearby the singular configuration allows a good path following and a robotized gesture accuracy. The choice of compression techniques for image transmission enables a compromise between flow and quality. These combined approaches, for robotics and image processing, enable the medical specialist to better control the remote ultrasound probe holder system and to receive stable and good quality ultrasound images to make a diagnosis via any type of communication link from terrestrial to satellite. Clinical tests have been performed since April 2003. They used both satellite or Integrated Services Digital Network lines with a theoretical bandwidth of 384 Kb/s. They showed the tele-echography system helped to identify 66% of lesions and 83% of symptomatic pathologies. PMID- 15787008 TI - Telemedicine diffusion in a developing country: the case of India (March 2004). AB - Telemedicine (health-care delivery where physicians examine distant patients using telecommunications technologies) has been heralded as one of several possible solutions to some of the medical dilemmas that face many developing countries. In this study, we examine the current state of telemedicine in a developing country, India. Telemedicine has brought a plethora of benefits to the populace of India, especially those living in rural and remote areas (constituting about 70% of India's population). We discuss three Indian telemedicine implementation cases, consolidate lessons learned from the cases, and culminate with potential researchable critical success factors that account for the growth and modest successes of telemedicine in India. PMID- 15787009 TI - Analysis of information and communication needs in rural primary health care in developing countries. AB - This article presents three studies dealing with information and communication needs in rural primary health care from Peru and Nicaragua. Results show that primary health-care systems in rural areas of developing countries are very inefficient. Among the main reasons we found factors related to communication infrastructure, information sharing, and continuous training of health professionals. We conclude that telemedicine systems can improve this situation, but the lack of infrastructures, low income levels, and other conditions, impose strong limits to the introduction of new technologies. The main conclusion is that differences in needs and conditions between developing countries and industrialized ones force to use different solutions and approaches. This article presents some proposals on technology requirements and how to deal with the use of telemedicine in rural areas of developing countries. These proposals can be useful to all kind of actors (national public administrations, multilateral institutions, industry, academy, civil society, etc.) in order to promote really relevant and sustainable proposals in telemedicine for rural regions of developing countries. PMID- 15787011 TI - An integrated computer supported acquisition, handling, and characterization system for pigmented skin lesions in dermatological images. AB - This paper describes an integrated prototype computer-based system for the characterization of skin digital images. The first stage includes an image acquisition arrangement designed for capturing skin images, under reproducible conditions. The system processes the captured images and performs unsupervised image segmentation and image registration utilizing an efficient algorithm based on the log-polar transform of the images' Fourier spectrum. Border- and color based features, extracted from the digital images of skin lesions, were used to construct a classification module for the recognition of malignant melanoma versus dysplastic nevus. Different methods, drawn from the fields of artificial intelligence (neural networks) and statistical modeling (discriminant analysis), were used in order to find the best classification rules and to compare the results of different approaches to the problem. PMID- 15787010 TI - Airmed-cardio: a GSM and Internet services-based system for out-of-hospital follow-up of cardiac patients. AB - A platform built around three information entities (patient, health-care_agent, and central_station) was designed to enable patients with chronic heart disease (in stable condition; emergency situations were excluded deliberately) to complete specifically defined protocols for out-of-hospital follow-up and monitoring. The patients belonged to one of four specific risk groups: arterial hypertension, malignant arrhythmias, heart failure, and postinfarction rehabilitation. They were provided with portable recording equipment and a cellular phone that supported data transmission [electrocardiogram (ECG)] and wireless application protocol (WAP) (remaining parameters and ad hoc questionnaires). The central station was an automatized platform, with no human operator. The information received was organized chronologically in patient folders. The health-care_agents had continuous and secure access to the patient folders, through tools based on the world wide web and WAP, and to short messages sent by their patients. A pilot project was conducted with 89 patients (mean length of participation: 50.1 days). A total of 2168 ECGs (mean duration transmission = 2 min/30 s; network errors < 0.1%) and 4011 short messages (none lost, in 95% of cases 30 s < delay < 1 min) were transmitted; 6083 WAP sessions (mean duration = 3 min 11 s; network failures < 0.1%) were The functionality of the platform was also evaluated, analyzing the subjective component of usability, showing the evolution of patient acceptance over time. PMID- 15787012 TI - Database design and implementation for quantitative image analysis research. AB - Quantitative image analysis (QIA) goes beyond subjective visual assessment to provide computer measurements of the image content, typically following image segmentation to identify anatomical regions of interest (ROIs). Commercially available picture archiving and communication systems focus on storage of image data. They are not well suited to efficient storage and mining of new types of quantitative data. In this paper, we present a system that integrates image segmentation, quantitation, and characterization with database and data mining facilities. The paper includes generic process and data models for QIA in medicine and describes their practical use. The data model is based upon the Digital Imaging and Communications in Medicine (DICOM) data hierarchy, which is augmented with tables to store segmentation results (ROIs) and quantitative data from multiple experiments. Data mining for statistical analysis of the quantitative data is described along with example queries. The database is implemented in PostgreSQL on a UNIX server. Database requirements and capabilities are illustrated through two quantitative imaging experiments related to lung cancer screening and assessment of emphysema lung disease. The system can manage the large amounts of quantitative data necessary for research, development, and deployment of computer-aided diagnosis tools. PMID- 15787013 TI - An evaluation of contrast enhancement techniques for mammographic breast masses. AB - The main aim of this paper is to propose a novel set of metrics that measure the quality of the image enhancement of mammographic images in a computer-aided detection framework aimed at automatically finding masses using machine learning techniques. Our methodology includes a novel mechanism for the combination of the metrics proposed into a single quantitative measure. We have evaluated our methodology on 200 images from the publicly available digital database for screening mammograms. We show that the quantitative measures help us select the best suited image enhancement on a per mammogram basis, which improves the quality of subsequent image segmentation much better than using the same enhancement method for all mammograms. PMID- 15787015 TI - Motion compensated lossy-to-lossless compression of 4-D medical images using integer wavelet transforms. AB - This paper proposes a method for progressive lossy-to-lossless compression of four-dimensional (4-D) medical images (sequences of volumetric images over time) by using a combination of three-dimensional (3-D) integer wavelet transform (IWT) and 3-D motion compensation. A 3-D extension of the set-partitioning in hierarchical trees (SPIHT) algorithm is employed for coding the wavelet coefficients. To effectively exploit the redundancy between consecutive 3-D images, the concepts of key and residual frames from video coding is used. A fast 3-D cube matching algorithm is employed to do motion estimation. The key and the residual volumes are then coded using 3-D IWT and the modified 3-D SPIHT. The experimental results presented in this paper show that our proposed compression scheme achieves better lossy and lossless compression performance on 4-D medical images when compared with JPEG-2000 and volumetric compression based on 3-D SPIHT. PMID- 15787014 TI - Virtual endoscopic visualization of the colon by shape-scale signatures. AB - We developed a new visualization method for virtual endoscopic examination of computed tomographic (CT) colonographic data by use of shape-scale analysis. The method provides each colonic structure of interest with a unique color, thereby facilitating rapid diagnosis of the colon. Two shape features, called the local shape index and curvedness, are used for defining the shape-scale spectrum. When we map the shape index and curvedness values within CT colonographic data to the shape-scale spectrum, specific types of colonic structures are represented by unique characteristic signatures in the spectrum. The characteristic signatures of specific types of lesions can be determined by use of computer-simulated lesions or by use of clinical data sets subjected to a computerized detection scheme. The signatures are used for defining a two-dimensional color map by assignment of a unique color to each signature region. The method was evaluated visually by use of computer-simulated lesions and clinical CT colonographic data sets, as well as by an evaluation of the human observer performance in the detection of polyps without and with the use of the color maps. The results indicate that the coloring of the colon yielded by the shape-scale color maps can be used for differentiating among the chosen colonic structures. Moreover, the results indicate that the use of the shape-scale color maps can improve the performance of radiologists in the detection of polyps in CT colonography. PMID- 15787016 TI - Volume manipulations for simulating bone and joint surgery. AB - Bone and joint surgery is widely used in orthopedic, oral, and maxillofacial, and dental and plasty departments to correct bone and joint pathology such as bone and joint tumors and fractures, and skeletal morphological deformities. This article presents a voxel structure to represent topologically and geometrically correct surfaces and algorithms to accurately compute intersections of tool swept surfaces with bones based on this voxel structure. This article then presents various volume manipulation algorithms to operate on virtual bones, bone grafts, and prostheses for bone and joint surgery simulations. A complicated knee arthroplasty illustrates the practicality and versatility of the proposed method. PMID- 15787017 TI - The clinical nurse leader: new in name only. PMID- 15787018 TI - Debate regarding interpretation of HESI exit examination scores. PMID- 15787019 TI - Evidence-based nursing education: myth or reality? AB - This article explores the concept of evidence-based nursing education. Because nurse educators incorporate evidence-based practice as a basic tenet of their programs, they assume nursing education itself is evidence based. Nursing education has a body of knowledge on which nurse educators base teaching, educational strategies, and curricular designs, but most of this knowledge is tacit, experiential, and based on practice. This knowledge relates to the art of teaching in nursing and can warrant the practice of nurse educators. However, research is also necessary to demonstrate the effectiveness of teaching approaches and strategies. Nurse educators need to develop the science of nursing education through qualitative and quantitative research, to add to the tacit knowledge underpinning nursing education strategies. When the science of nursing education is adequately developed through rigorous research, we will truly be able to say that nursing education is evidence based. Until then, it may be only a myth. PMID- 15787020 TI - Multiple role adaptation among women who have children and re-enter nursing school in Taiwan. AB - This study assessed multiple role adaptation within maternal and student roles among female RNs who had children and returned to school for baccalaureate degrees in Taiwan. Using Roy's Adaptation Model as the theoretical framework, relationships were explored among demographic (number of children, age of youngest child, employment status), physical (sleep quality, health perception, activity), and psychosocial factors (self-identity, role expectation, role involvement, social support) and multiple role adaptation (role accumulation). The sample included 118 mother-students who had at least one child younger than age 18 and who were studying in nursing programs in Taiwan. The highest correlation was found between activity and role accumulation followed by significant correlations between sleep quality, health perception, maternal role expectation, and age of youngest child and role accumulation. In regression analyses, the complete model explained 46% of the variance in role accumulation. Implications for education and future research are identified. PMID- 15787021 TI - Participatory inquiry with a colleague: an innovative faculty development process. AB - Clinical evaluation is central to the aims of nursing education; however, little has been written about the actual evaluative practices of nurse educators and the sources of influence on those practices. In this article, we describe our experience as co-investigators into the evaluation practices of one of us (J.A.J.) and overview some of the challenges and opportunities of participatory inquiry. We describe how J.A.J.'s involvement in this research project encouraged her to explore her evaluative practices in a meaningful way, which moved her to a place of deeper understanding about her work and transformed her as a nurse educator. As a result of this study, we are convinced of the necessity for a faculty development process whereby colleagues commit to a critical inquiry process focused on identifying each other's evaluative practices and sources of influence on those practices. In addition, schools of nursing can encourage faculty participation in peer development by adopting an expanded view of scholarship in which the disciplined examination of one's evaluative practices is accepted as evidence of the scholarship of teaching. PMID- 15787022 TI - A pilot study to investigate the effect of a simulation strategy on the clinical decision making of midwifery students. AB - The actual effect of the use of simulations on clinical decision making is inconclusive. This pilot study used a posttest design to determine the effect of a simulation strategy on the clinical decision-making process of midwifery students. Thirty-six graduate diploma students volunteered and were randomly assigned to two groups, with the experimental group receiving two simulation sessions (normal labor and physiological jaundice), and the control group receiving the two usual lectures. The main findings were that students who received the simulation strategy collected more clinical information, revisited collected clinical information less, made fewer formative inferences, reported higher confidence levels, and for the posttest normal labor simulation, reached a final decision more quickly. Such effects are reasonable for this type of intervention with the existent variability in each group. Further research with a larger sample size and more rigorous data collection strategies is required. PMID- 15787023 TI - What do high school guidance counselors really think of nursing? AB - With the increasing nursing shortage, it is imperative that nursing programs attract and retain students who will be successful in becoming competent, caring practitioners. High school guidance counselors can influence the career choices of students. To do so, these professionals must have an accurate understanding of the attributes that will facilitate a career in nursing, as well as the changing roles of nurses. This study determined the current perceptions of high school guidance counselors regarding nursing. Overall, guidance counselors had realistic perceptions of nursing and identified personal and cognitive attributes of students that would promote success in nursing. However, nurses involved in the recruitment of students need to ensure that guidance counselors have more accurate perceptions of the changing role of nurses, opportunities for practice outside hospital settings, and the importance of problem-solving abilities and leadership skills for nurses. PMID- 15787024 TI - Exploring a model to evaluate levels of reflection in baccalaureate nursing students' journals. AB - The purpose of this study was to use Mezirow's model to identify reflective levels among baccalaureate nursing students. This study examined the question, "Tb what extent and at what level is reflection-on-practice identified in the journals of junior baccalaureate nursing students?" Twenty nursing students wrote journals at three points during a health assessment course. This descriptive, non experimental study used a classification system to quantify the amount of reflection present. Eighty percent of students were able to reach the higher levels of reflection in at least one journal, but 20% had no evidence of higher level reflective thought in any journal. Sixty-three percent of journals had entries at the higher levels of reflection. However, of all journal entries, 18% were at higher levels, and 82% were at lower levels. These findings are consistent with other nursing studies using Mezirow's model. Mezirow's levels of reflection are a useful tool to evaluate the level of student reflection in journals. PMID- 15787025 TI - Teaching epidemiology using WebCT: application of the seven principles of good practice. AB - Web-based teaching was strategically used to supplement classroom instruction in an epidemiology course for RN-to-BSN students. This article investigates how the "Seven Principles for Good Practice" were applied as a guide when teaching this online course and discusses this innovative teaching strategy with the aim of advancing pedagogical strategies from the traditional, teacher-centered paradigm to an active, learner-centered paradigm. PMID- 15787026 TI - Service learning: an adjunct to therapeutic communication and critical thinking skills for baccalaureate nursing students. PMID- 15787027 TI - Community health nursing in a virtual setting. PMID- 15787028 TI - Proceedings of the Third Biennial National Conference on Health Issues in the Arab-American Community. Dearborn, Michigan, USA. October 24-25, 2003. PMID- 15787029 TI - The problem of cancer: lung cancer as a paradigm--keynote address summary. PMID- 15787030 TI - Cancer databases in the Arab world. PMID- 15787031 TI - Cancer epidemiology in Arab Americans and Arabs outside the Middle East. PMID- 15787032 TI - An update on recent cancer trends in Lebanon. PMID- 15787033 TI - Childhood cancer among Arab Americans in southeast Michigan. PMID- 15787034 TI - Measurement of the knowledge, attitudes, and beliefs of Arab-American adults toward cancer screening and early detection: development of a survey instrument. PMID- 15787036 TI - Trends of body mass index (BMI) by age and sex percentile among Arab-American fifth graders in southeastern Michigan schools. PMID- 15787037 TI - Obesity and cardiovascular disease risk factors are ethnicity based: a study of women of different ethnic backgrounds in southeastern Michigan. PMID- 15787038 TI - Diabetes and the metabolic syndrome in Arab Americans: a community-based sample. PMID- 15787039 TI - Cardiovascular risk factors in an Arab-American population in southeastern Michigan. PMID- 15787040 TI - The epidemiology of diabetes and its risk factors among Chaldean Americans. PMID- 15787042 TI - Science and the community: a collaborative model for integration of research within the community. PMID- 15787043 TI - Environmental impacts on Arab Americans in metropolitan Detroit. PMID- 15787044 TI - Environmental justice and environmental health: focus group assessments of the perceptions of Arab Americans in metro Detroit. PMID- 15787045 TI - Environmental disease burden in Arab-American communities in the Detroit area: prevalence and severity. PMID- 15787046 TI - Costs of environmentally attributable diseases in the Arab-American community in the Detroit area. PMID- 15787048 TI - Trends in tobacco use among Arab/Arab-American adolescents: preliminary findings. PMID- 15787047 TI - Culturally sensitive smoking cessation intervention program redesign for Arab American youth. PMID- 15787049 TI - Relationship of parental tobacco use, peer influence, self-esteem, and tobacco use among Yemeni-American adolescents: mid-range theory testing. PMID- 15787050 TI - Health and disease in a changing Arab world 2000/2025/2050: global, environmental, and climate change and emerging diseases. PMID- 15787051 TI - Major health problems in North Africa. PMID- 15787052 TI - Major health issues in Jordan. PMID- 15787053 TI - The health status of the Palestinian Arab community in Israel in relation to the Jewish community of Israel. PMID- 15787054 TI - Palestinian health system after three years of the Intifada--survival, development, or both? PMID- 15787055 TI - Mental health situation in Morocco: problems and perspectives. PMID- 15787056 TI - Status of nursing in the Arab world. PMID- 15787057 TI - Advancing the health of Arab Americans: key points to obtaining resources and establishing programs focused on special populations. PMID- 15787058 TI - Refugee hurdles, caretaker challenges, and mutual openings--keynote speaker. PMID- 15787059 TI - The psychosocial rehabilitation approach in treating torture survivors. PMID- 15787060 TI - "Each one is a doctor for herself": Ramadan fasting among pregnant Muslim women in the United States. PMID- 15787061 TI - The effects of stigma consciousness on mental health. PMID- 15787062 TI - The aftermath of the Persian Gulf War: mental health issues among Iraqi-American veterans. PMID- 15787063 TI - Depression and medical health complaints in a group of Arab-American women. PMID- 15787064 TI - Reactions of Arab-American groups to the September 11, 2001 terrorist attacks. PMID- 15787065 TI - Social relations and mental health among Arab-American elders. PMID- 15787066 TI - High-effort active coping and health outcomes among older Arab immigrants. PMID- 15787067 TI - Examination of the impact of acculturation, stress, and religiosity on mental health variables for second-generation Arab Americans. PMID- 15787068 TI - [Epidemiology of infection and sepsis in intensive care unit patients]. AB - Since 1992, epidemiological and clinical studies have classified severe infections into three categories: sepsis, severe sepsis and septic shock. Microbiological documentation is not always provided. We used a different approach, focusing on the infection itself, whether or not it is microbiologically documented or associated with sepsis. In an international prospective cohort study, all patients admitted to the participating units from May 1997 to May 1998 were followed until hospital discharge. Twenty-eight intensive care units (ICU) in eight countries enrolled 14,364 patients. Of these, 6011 stayed in the ICU for less than 24 hours and 8353 for more than 24 hours. Overall, 3034 infectious episodes were recorded at ICU admission (crude incidence rate 21.1%). Among patients hospitalized for more than 24 hours, 1581 infectious episodes occurred in the ICU (crude incidence rate 18.9%), including 713 cases (45%) in patients who were already infected at ICU admission. These rates varied among the ICUs. Respiratory, gastrointestinal, urinary tract and primary bloodstream infections represented about 80% of all infections. Hospital-acquired and Intensive Care Unit-acquired infections were more frequently microbiologically documented than community-acquired infections (71% and 86%, respectively, vs 55%). About 28% of all infections were associated with sepsis, 24% with severe sepsis and 30% with septic shock (18% were not classified). Crude in-hospital mortality rates ranged from 16.9% in uninfected patients to 53.6% in patients who were both infected at the time of ICU admission and subsequently acquired an infection during the ICU stay. The in-hospital mortality rate increased with severity, from 20% for sepsis to 40% for severe sepsis and 60% for septic shock, but also depended on the origin of infection (community vs hospital/ICU). Crude incidence rates of ICU infection were high, varying among ICUs and patient subsets. Thus, vital outcome depends not only on the severity of sepsis but also on the characteristics of the infection. PMID- 15787069 TI - [Mortality and morbidity of low-birth-weight infants. Current prognosis and future perspectives]. AB - In a retrospective epidemiologic study conducted from January 1996 to December 2002, 2555 low-birth-weight infants (less than 2500 g) were admitted to the neonatal intensive care unit of E Herriot Hospital in Lyon, France. Very low birth weight (795 infants) was defined as a birth weight below 1500 g. The total mortality rate among these latter infants was 8%. The mortality rate was less than 25% among infants with a gestational age of at least 25 weeks. Neonatal morbidity mainly consisted of respiratory distress syndrome and cerebral hemorrhage. Sequelae at age 2 years (corrected) affected less than 10% of surviving in very-low-birth-weight infants. Gestational age is thus an important predictor of survival and functional outcome in very-low-birth-weight infants. PMID- 15787070 TI - [New concepts in epidemiological surveillance in French army]. AB - Epidemiological surveillance within the French Armed Forces has had to take into account various changes in infectious diseases in recent years. The French Armed Forces are encountering new hazards, such as the spread of HIV infection, Plasmodium falciparum chemoresistance, and outbreaks of emerging diseases. Bioterrorism, industrial and occupational hazards are added concerns. For these reasons, the French Military Medical Service has introduced a new concept based on permanent epidemiological surveillance of communicable diseases. This is completed by a real-time spatial surveillance designed to detect very rapidly potential communicable diseases or new emerging diseases. This epidemiological system, based on data modeling, enhances the medical information available to staff commands before deployment to new areas. PMID- 15787071 TI - [The visual pathways, from anatomical MRI to physiological with (f)MRI and tractography with diffusion tensor MRI (DTMRI)]. AB - Advances in MRI technology have led to a better knowledge of visual pathways (1984-2004), with a new descriptive anatomy and functional model. The authors first describe the technical development of MRI over the last thirty years, then describe and illustrate the new descriptive anatomy. Cephalic MRI reveals brain structures that were previously invisible, on different encephalic planes, in the optic pathways, horizontally from the cornea to the calcarin fissure (neuro ocular plane (NOP), oblique trans-hemispheric neuro-ocular (OTNOP) and neuro-opto tractal planes (NOTP)), in their orthogonal orientation upon the oculomotor pathways: head and axonal optic nerve pack (visual deutoneurons in their meninges), optic tracts, lateral geniculate bodies, optic radiations and the calcarian fissure. Comparative anatomy with the rhesus macaque is mentioned. Functional neuroanatomy (physiology) benefits from cine-MRI for ocular motricity (OD MRI), growth by the observation of myelinization in children, blood and CSF circulation by MR angiography, local blood volumes by perfusion imaging, neuronal quantification with inflammation or myelin regeneration by spectroscopy (MRS), brain mapping by functional MR ((f)MRI) measuring local CBF enhancement by paradigmatic stimulations. The recent functional imaging method, tractography (or diffusion tensor MRI (DTMRI)), using diffusion MRI techniques, natural vector calculations with diffusion tensor and software power for morphological and statistical directional results, represents the direction of projection, association and commissural white matter tracts. Normal examples are shown and some common clinical consequences are discussed. PMID- 15787072 TI - [Treatment by embolization of small uterine myomata]. AB - We examined the effectiveness of uterine artery embolization (UAE) as a primary treatment for small symptomatic myomatas (<50 mm diameter). One hundred patients aged from 29 to 68 years underwent UAE between 01/01/99 and 12/31/2002. Efficacy was judged by physical examination and imaging studies at 6 to 12 months. All the patients were evaluated. Treatment failed in five cases, while symptoms resolved in all the other patients. Five patients became pregnant. The main complication was amenorrhea. No serious adverse effects were observed. The efficacy and safety of this method make it a possible preventive treatment for pre-symptomatic myomata. If these findings are confirmed, they will open a new era in the treatment of fibroids. PMID- 15787073 TI - [Obligatory information of patients with rheumatic illness (300 cases)]. AB - Since 1997, legal judgments in France have revolutionized the concept of patient information. Medical information must now be clear and include all serious risks of investigations and therapies. We evaluated patients' opinions after their medical examination and scrutiny of information cards prepared by the French Society of Rheumatology. A questionnaire was used to assess the effectiveness and comprehension of the cards, and their potential to induce anxiety. Written informed consent was provided by the majority of patients. The medical information was well received, especially in oral form, and was considered comprehensible but relatively stressful. Anxiety was significantly higher among female patients and patients who had never had medical examinations or procedures. PMID- 15787074 TI - About a boy. Carter Lee's story put a face on one of the medical profession's biggest issues: access. PMID- 15787075 TI - Loss prevention. The way things should go. PMID- 15787076 TI - Arkansas' inpatient performance improves. Cardiac care still below national averages. PMID- 15787077 TI - Radical prostatectomy and postoperative radiation in patients with adenocarcinoma of prostate of intermediate and high risk for recurrence. AB - OBJECTIVE: To report on the outcome of patients with intermediate and high risk of recurrence who underwent radical prostatectomy (RP). METHODS: Eighty-five consecutive patients categorized as intermediate (17.5%) and high risk (82.5%) of failure after definitive therapy for carcinoma of prostate according to the National Comprehensive Cancer Network (NCCN) underwent RP between 1989 and 1997. Median preoperative PSA was 26 ng/ml (range 15 ng/ml-91 ng/ml). Fifty-nine patients (70%) received three months neoadjuvant hormone therapy. Thirty-six patients (42%) underwent early (three to four months after RP) adjuvant radiation for pT3 disease and/or positive surgical margins. RESULTS: The median follow-up was 58 months (range 12-104 months). There was no difference in the biochemical recurrence rate between the intermediate and high-risk group of patients. The overall relapse rate was 33%. Cancer-specific mortality was 3.5%. Patients with T1c tumors had a significantly lower biochemical recurrence rate (bRR) (7%) compared to palpable tumors (p = 0.03). Age above 65 was a significant negative prognostic factor with respect to biochemical recurrence (p = 0.01). Adjuvant radiation was associated with biochemical recurrence rates of 25% vs. 40% in patients who were not radiated (p = 0.05). CONCLUSIONS: In the intermediate and high-risk groups of patients with nonpalpable prostate cancer, RP and adjuvant RT may provide a biochemical recurrence-free rate (bRFR) comparable to that reported in other series with RP alone on patients in the low-risk groups. We encourage the multimodality treatment approach incorporating adjuvant postoperative radiation in these patients. PMID- 15787078 TI - Rare auditory-electophysiology finding in Wilson's disease. AB - Wilson's disease is a rare genetic disease involving the malabsorption of copper by the body. The most common characteristic sign is the presence of Kayser Fleischner ring surrounding the cornea. Other systemic and motor signs have been documented as well as MRI changes within the brain and brainstem. This rare case illustrates the potential importance of audiometric assessment for patients with Wilson's disease who complain of hearing loss, tinnitus and intra-aural pressure. Unilateral findings were significant for retrocochlear neural transmission delays. PMID- 15787079 TI - Eliminating biased language: a goal for everyone. PMID- 15787080 TI - The publication of the proposed new regulations for the ESRD program: an historic event for nephrology nursing. PMID- 15787081 TI - A clinical review of antibody-mediated pure red cell aplasia. AB - Pure red cell aplasia (PRCA) is a rare hematologic disorder characterized by severe anemia from selectivefailure of the bone marrow to produce mature red blood cells. This disorder can be congenital or acquired from various causes. The development of PRCA in patients with chronic kidney disease receiving recombinant erythropoietin (rHuEPO) for renal anemia has increased in recent years. This review will enable nurses to recognize the pathogenesis, clinical manifestations, and therapeutic options for rHuEPO-induced PRCA. PMID- 15787082 TI - State laws and regulations specific to dialysis: an overview. PMID- 15787083 TI - Incidence and influencing factors associated with exit site infections in temporary catheters for hemodialysis and apheresis. AB - The authors examined the incidence and influencing actors of exit site infections (ESIs) in patients using dual and triple lumen dialysis catheters. The prospective, randomized study was conducted at 11 centers. Data were analyzed on 473 patients randomized to receive 235 dual lumen catheters (DLCs) or 238 triple lumen catheters (TLCs). RESULTS: ESIs occurred in 10 patients (2.1%). The number of intertreat-ment uses (p = 0.006), total number of uses (p = 0.009), and catheter placement duration (p = 0.03) were significantly associated with incidence of ESI in both treatment groups. Although patients with TLCs had a significantly higher inter-treatment use than patients with DLCs (p = .04), no sgnificant difference in the incidence of ESI was found (p = 0.11). Diabetic status and catheter insertion site were not found to be significantly related to ESI ( p= 0.09 and p = 0.54, respectively), although a trend toward significance was noted for diabetic status. CONCLUSIONS: Stringent exit site care by specially trained staff and shorter catheter duration most likely contributed to the low incidence of ESIs. Other influencing factors included numbers of inter-treatment uses, total number of uses, and catheter placement duration. PMID- 15787084 TI - Medicare survey of the kidney transplant unit: are you prepared? AB - Facilities or hospitals must be approved to partiapate in Medicare's End Stage Renal Disease (ESRD) Program, and this includes kidney transplant programs. This article provides an overview of a Basic Survey and what requirements are outlined in the ESRD conditions for Coverage to achieve and maintain Medicare approval and funding. PMID- 15787085 TI - Evaluating quantitative research reports. AB - As a novice reviewer, it is often difficult to trust your evaluation of a research report. You may feel uncertain in your interpretations. These are common concerns and can be remedied by reading and discussing research reports on research listservs, through journal clubs, or with other nephrology nurses. Practice using the criteria for research report evaluation and you too can perfect critiquing a research report! PMID- 15787086 TI - Evaluating traditional iron measures and exploring new options for patients on hemodialysis. AB - Despite the use of recombinant human erythropoietin (rHuEPO), anemia remains a major problem in patients with chronic kidney disease and end stage renal disease. In addition, these disease states are often complicated by iron deficiency, especially in patients on chronic hemodialysis. Adequate iron supplies are necessary for normal erythropoiesis to occur and for the patient to effectively respond to rHuEPO treatment. Therefore, it is important for the nephrology nurse to have reliable iron test data in order to accurately monitor a patient's iron status. The two primary tests used to assess iron status are transferrin saturation and serum ferritin level. Both tests are easy to perform and are cost-efficient but have limitations and are highly variable under certain physiologic condtions, such as inflammation and infection. Due to these limitations, research has focused on alternative methods of iron assessment, with some newer techniques being more effective than others. Although nephrology nurses depend on transferrin saturation and serum ferritin levels to indicate iron status, they should recognize the associated restrictions and treat the patient based on a complete evaluation. PMID- 15787087 TI - Functional ability of patients on dialysis: the critical role of anemia. AB - Maintining hemoglobin (Hb) levels in the range (11 to 12 g/dL) recommended by the National Kidney Foundation's Kidney Disease Quality Initiative (NKF/KDOQI) has been shown to significantly improve functional ability in patients on dialysis. The combination of partial anemia correction and an individualized exercise program yields additional incremental benefits. Nurses fulfill a crucial role in ensuring that appropriate Hb levels are maintained while at the same time encouraging patients to improve their physical conditioning and functional status. PMID- 15787088 TI - The challenge of wound healing in patients with chronic kidney disease. PMID- 15787089 TI - Which credentials should nursing adopt as standard? NP should be the only credential used in clinical practice. PMID- 15787090 TI - Which credentials should nursing adopt as standard? Preserve the CNS as it was meant to be recognized. PMID- 15787091 TI - Which credentials should nursing adopt as standard? APN and APRN are the appropriate credentials. PMID- 15787092 TI - Carnitine therapy... Let the patients benefit! PMID- 15787093 TI - Dialysis decisions: should everyone be dialyzed? PMID- 15787095 TI - Plasma TGF-beta1-related survival of postmenopausal metastatic breast cancer patients. AB - A pilot study was conducted to assess whether plasma levels of transforming growth factor-beta1 (TGF-beta1) might facilitate biological subgrouping of postmenopausal metastatic breast cancer patients, and, accordingly, its applicability in clinical oncology. This study included 29 postmenopausal metastatic breast cancer patients. Plasma TGF-beta1 levels were detected by enzyme-linked immunosorbent assay (ELISA). Estrogen and progesterone receptors were assayed by radioligand binding, in accordance with the recommendation of the EORTC. Concentrations of 17-beta estradiol were determined by using ELISA microwell method (DIALAB). Overall survival was followed for 24 months for each individual patient. Stratification of the patients by ER/PR status showed that 14 patients with estrogen receptor-negative, progesterone receptor-negative carcinomas displayed a statistically significant increase in plasma TGF-beta1 levels when compared to plasma TGF-beta1 levels of 6 patients with ER-positive, PR-positive carcinomas (P=0.04). In this study, 7 out of 14 patients with negative receptors' status had no plasma TGF-beta1 values overlapping with patients having positive receptors' status. The TGF-beta1 cut-off value was defined as the highest plasma TGF-beta1 level of ER-positive, PR-positive patients: 3.28 ng/ml. This plasma TGF-beta1 cut-off value defined low-risk subgroup of 19 patients (< or = 3.28 ng/ml) and high-risk subgroup of 10 patients (> 3.28 ng/ml) (P=0.047). Plasma TGF-beta1-related survival was independent of the classical prognostic factors of metastatic breast cancer. Accordingly, a clinical significance of elevated plasma TGF-beta1 levels may be suggested. PMID- 15787094 TI - Constitutive expression of growth regulated oncogene (gro) in human colon carcinoma cells with different metastatic potential and its role in regulating their metastatic phenotype. AB - The purpose of this study was to examine the expression and functional significance of the growth-regulated oncogene (gro) family in human colon carcinoma growth and metastasis. We examined constitutive expression of CXCL1 (gro-alpha), CXCL2 (gro-beta), CXCL3 (gro-gamma) and their receptor, CXCR2 in human colon carcinoma cells with different metastatic potentials. Non-metastatic and low metastatic cells expressed lower levels of CXCL1 and CXCR2 mRNA and protein as compared to high metastatic colon carcinoma cells. No difference in CXCL2 and CXCL3 mRNA expression levels was observed. Colon carcinoma cells expressing higher levels of CXCL1 exhibit increased proliferation and invasive potential. Furthermore, exogenous addition of recombinant human CXCL1 significantly enhanced the proliferation and invasiveness of colon carcinoma cells. Furthermore, treatment of KM12C cells with exogenous CXCL1 enhanced their invasiveness. Neutralizing antibody to CXCL1 in combination with antibody to CXCR2 inhibited highly metastatic KM12L4 (high CXCL1 expressor) cell proliferation. These data demonstrate that the constitutive expression of CXCL1 and its receptor CXCR2 is associated with metastatic potential and modulates colon cancer cell proliferation and an invasive phenotype. PMID- 15787097 TI - MedPAC recommends specialty moratorium extension, FY'06 -0.4 percent market basket update, pay for performance/IT standards. PMID- 15787096 TI - Specific amino acid restriction inhibits attachment and spreading of human melanoma via modulation of the integrin/focal adhesion kinase pathway and actin cytoskeleton remodeling. AB - We had previously found that selective restriction of amino acids inhibits invasion of human A375 melanoma. Integrins, cell surface receptors for the components of extracellular matrix (ECM), are activated during cell adhesion and spreading, and initiate signaling pathways that control growth and invasion of tumor cells. We examined the effect of tyrosine (Tyr) and phenylalanine (Phe), methionine (Met) or glutamine (Gln) restriction on attachment and spreading of A375 and MeWo melanoma cell lines on fibronectin and laminin. In A375 cells, restriction of Tyr/Phe or Met inhibited attachment to and spreading on laminin and fibronectin, inhibited alpha3 and alpha4 integrin expression, and inhibited accumulation of FAK-Tyr397 and F-actin at leading edges of cell protrusions. Tyr/Phe restriction also inhibited attachment-induced autophosporylation of FAK Tyr397. In MeWo cells, the order of inhibition by amino acid restriction on cell attachment and spreading was as follows: Gln > Tyr/Phe > Met. Restriction of Gln reduced alpha5 integrin expression. All amino acid restrictions similarly inhibited phosphorylation of FAK-Tyr397, FAK-Tyr577, FAK-Tyr861 and paxillin Tyr31. Gln restriction exhibited the strongest inhibition of actin cytoskeleton remodeling during the cell spreading. The present study reveals that specific amino acid restriction inhibits attachment and spreading of melanoma via inhibition of specific integrin expression, inhibition of integrin-mediated FAK phosphorylation, and modulation of actin cytoskeleton remodeling. These data provide additional understanding of the mechanism by which specific amino acid restriction controls invasion and migration of melanoma. PMID- 15787099 TI - Studies show Medicare Rx benefit savings. PMID- 15787098 TI - CMS releases hospital OPPS, inpatient psych PPS and Physician Fee Schedule. PMID- 15787100 TI - Health spending growth increases, hospital inpatient spending declines. PMID- 15787101 TI - Escalating budget crisis threatens healthcare's future. PMID- 15787102 TI - Public opposes hospital payment cuts. PMID- 15787103 TI - ICUs tighten belts on blood glucose levels. PMID- 15787104 TI - Clearing a path for new cardiac markers. PMID- 15787105 TI - On client billing, a voice in the wilderness. PMID- 15787106 TI - Pilot points way to speedier cancer surveillance. PMID- 15787107 TI - Be seen and be heard. PMID- 15787108 TI - Reticulocyte counting. PMID- 15787109 TI - Replacing an old LIS. PMID- 15787110 TI - Lab automation--what a long, lively trip it's been. PMID- 15787111 TI - Maryland lab case--little to learn, lots to remember. PMID- 15787112 TI - 'They know things I don't know'. PMID- 15787113 TI - Pathology CME--moving from general to specific. PMID- 15787114 TI - The next picture show. New products are encountering a market ripe for digital imaging. PMID- 15787115 TI - Capitol scan. PMID- 15787116 TI - The professional in autumn. PMID- 15787117 TI - [Bronchopulmonary dysplasia (BPD) and CPAP use on newborn and premature infants. II. Management of CPAP respiration]. PMID- 15787118 TI - [Nursing measures in stem cell transplantation]. PMID- 15787119 TI - [Relations with shame feelings in nursing]. PMID- 15787120 TI - [Treatment of pain with nonopioid analgesics in childhood and adolescence -- new analgesics similar to "better" analgesics?]. PMID- 15787121 TI - [Travel pharmacy for children]. PMID- 15787122 TI - [Round Table: Ambulatory nursing]. PMID- 15787123 TI - [High sick leave in hospitals and nursing homes]. PMID- 15787124 TI - [EBDD Annual Report 2004: decline of drug-induced mortality -- concerns about increasing cannabis consumption by adolescents]. PMID- 15787125 TI - Emergency, shmergency! PMID- 15787126 TI - Are we ready for medical emergencies? PMID- 15787127 TI - The state of preparedness in Texas dental practices for medical emergency responses: a survey. PMID- 15787128 TI - Office emergency preparation and equipment. PMID- 15787130 TI - The use of oxygen and airway modalities in dental office medical emergencies. PMID- 15787129 TI - The pharmacology of recommended medical emergency drugs. PMID- 15787131 TI - Portable blood glucose testing meters in dental practice: a valuable medical emergencies adjunct. PMID- 15787132 TI - Selecting an automated cardiac defibrillator (AED) for the dental office: which is "best"? PMID- 15787133 TI - The psychology of money. AB - Money is an exchange of value. Your patient receives the value of health, well being, an improved appearance, and keeping teeth for a lifetime of comfort and function. These are life-enhancing benefits. You are receiving money as the patient's part of the exchange. Money is a moveable entity that flows from one person to another or from one business to another. Both you and your patients benefit from an equitable exchange of value. That occurs when you feel good about the money you receive for services rendered and your patient feels good about the investment they have made in you and in your care. Didn't Dr. Pankey teach us that? PMID- 15787134 TI - Dental artifacts--it was in the cards! PMID- 15787135 TI - Oral and maxillofacial pathology case of the month. Osteoma, associated with Gardner syndrome. PMID- 15787136 TI - Informed consent: it takes two to establish the doctor-patient relationship. PMID- 15787137 TI - Encoding strategy for maximum noise tolerance bidirectional associative memory. AB - In this paper, the basic bidirectional associative memory (BAM) is extended by choosing weights in the correlation matrix, for a given set of training pairs, which result in a maximum noise tolerance set for BAM. We prove that for a given set of training pairs, the maximum noise tolerance set is the largest, in the sense that this optimized BAM will recall the correct training pair if any input pattern is within the maximum noise tolerance set and at least one pattern outside the maximum noise tolerance set by one Hamming distance will not converge to the correct training pair. This maximum tolerance set is the union of the maximum basins of attraction. A standard genetic algorithm (GA) is used to calculate the weights to maximize the objective function which generates a maximum tolerance set for BAM. Computer simulations are presented to illustrate the error correction and fault tolerance properties of the optimized BAM. PMID- 15787138 TI - Stability criteria for unsupervised temporal association networks. AB - A biologically realizable, unsupervised learning rule is described for the online extraction of object features, suitable for solving a range of object recognition tasks. Alterations to the basic learning rule are proposed which allow the rule to better suit the parameters of a given input space. One negative consequence of such modifications is the potential for learning instability. The criteria for such instability are modeled using digital filtering techniques and predicted regions of stability and instability tested. The result is a family of learning rules which can be tailored to the specific environment, improving both convergence times and accuracy over the standard learning rule, while simultaneously insuring learning stability. PMID- 15787142 TI - Self-organizing learning array. AB - A new machine learning concept--self-organizing learning array (SOLAR)--is presented. It is a sparsely connected, information theory-based learning machine, with a multilayer structure. It has reconfigurable processing units (neurons) and an evolvable system structure, which makes it an adaptive classification system for a variety of machine learning problems. Its multilayer structure can handle complex problems. Based on the entropy estimation, information theory-based learning is performed locally at each neuron. Neural parameters and connections that correspond to minimum entropy are adaptively set for each neuron. By choosing connections for each neuron, the system sets up its wiring and completes its self-organization. SOLAR classifies input data based on the weighted statistical information from all the neurons. The system classification ability has been simulated and experiments were conducted using test-bench data. Results show a very good performance compared to other classification methods. An important advantage of this structure is its scalability to a large system and ease of hardware implementation on regular arrays of cells. PMID- 15787143 TI - Analysis of augmented-input-layer RBFNN. AB - In this paper we present and analyze a new structure for designing a radial basis function neural network (RBFNN). In the training phase, input layer of RBFNN is augmented with desired output vector. Generalization phase involves the following steps: (1) identify the cluster to which a previously unseen input vector belongs; (2) augment the input layer with an average of the targets of the input vectors in the identified cluster; and (3) use the augmented network to estimate the unknown target. It is shown that, under some reasonable assumptions, the generalization error function admits an upper bound in terms of the quantization errors minimized when determining the centers of the proposed method over the training set and the difference between training samples and generalization samples in a deterministic setting. When the difference between the training and generalization samples goes to zero, the upper bound can be made arbitrarily small by increasing the number of hidden neurons. Computer simulations verified the effectiveness of the proposed method. PMID- 15787144 TI - An energy function-based design method for discrete Hopfield associative memory with attractive fixed points. AB - An energy function-based autoassociative memory design method to store a given set of unipolar binary memory vectors as attractive fixed points of an asynchronous discrete Hopfield network (DHN) is presented. The discrete quadratic energy function whose local minima correspond to the attractive fixed points of the network is constructed via solving a system of linear inequalities derived from the strict local minimality conditions. The weights and the thresholds are then calculated using this energy function. If the inequality system is infeasible, we conclude that no such asynchronous DHN exists, and extend the method to design a discrete piecewise quadratic energy function, which can be minimized by a generalized version of the conventional DHN, also proposed herein. In spite of its computational complexity, computer simulations indicate that the original method performs better than the conventional design methods in the sense that the memory can store, and provide the attractiveness for almost all memory sets whose cardinality is less than or equal to the dimension of its elements. The overall method, together with its extension, guarantees the storage of an arbitrary collection of memory vectors, which are mutually at least two Hamming distances away from each other, in the resulting network. PMID- 15787145 TI - A recurrent neural network for solving nonlinear convex programs subject to linear constraints. AB - In this paper, we propose a recurrent neural network for solving nonlinear convex programming problems with linear constraints. The proposed neural network has a simpler structure and a lower complexity for implementation than the existing neural networks for solving such problems. It is shown here that the proposed neural network is stable in the sense of Lyapunov and globally convergent to an optimal solution within a finite time under the condition that the objective function is strictly convex. Compared with the existing convergence results, the present results do not require Lipschitz continuity condition on the objective function. Finally, examples are provided to show the applicability of the proposed neural network. PMID- 15787146 TI - Passivity-based neural network adaptive output feedback control for nonlinear nonnegative dynamical systems. AB - The potential clinical applications of adaptive neural network control for pharmacology in general, and anesthesia and critical care unit medicine in particular, are clearly apparent. Specifically, monitoring and controlling the depth of anesthesia in surgery is of particular importance. Nonnegative and compartmental models provide a broad framework for biological and physiological systems, including clinical pharmacology, and are well suited for developing models for closed-loop control of drug administration. In this paper, we develop a neural adaptive output feedback control framework for adaptive set-point regulation of nonlinear uncertain nonnegative and compartmental systems. The proposed framework is Lyapunov-based and guarantees ultimate boundedness of the error signals corresponding to the physical system states and the neural network weighting gains. The approach is applicable to nonlinear nonnegative systems with unmodeled dynamics of unknown dimension and guarantees that the physical system states remain in the nonnegatiye orthant of the state-space for nonnegative initial conditions. Finally, a numerical example involving the infusion of the anesthetic drug midazolam for maintaining a desired constant level of depth of anesthesia for noncardiac surgery is provided to demonstrate the efficacy of the proposed approach. PMID- 15787147 TI - Neural network adaptive control for nonlinear nonnegative dynamical systems. AB - Nonnegative and compartmental dynamical system models are derived from mass and energy balance considerations that involve dynamic states whose values are nonnegative. These models are widespread in engineering and life sciences and typically involve the exchange of nonnegative quantities between subsystems or compartments wherein each compartment is assumed to be kinetically homogeneous. In this paper, we develop a full-state feedback neural adaptive control framework for adaptive set-point regulation of nonlinear uncertain nonnegative and compartmental systems. The proposed framework is Lyapunov-based and guarantees ultimate boundedness of the error signals corresponding to the physical system states and the neural network weighting gains. In addition, the neural adaptive controller guarantees that the physical system states remain in the nonnegative orthant of the state-space for nonnegative initial conditions. PMID- 15787148 TI - Direct adaptive controller for nonaffine nonlinear systems using self-structuring neural networks. AB - A direct adaptive state-feedback controller is proposed for highly nonlinear systems. We consider uncertain or ill-defined nonaffine nonlinear systems and employ a neural network (NN) with flexible structure, i.e., an online variation of the number of neurons. The NN approximates and adaptively cancels an unknown plant nonlinearity. A control law and adaptive laws for the weights in the hidden layer and output layer of the NN are established so that the whole closed-loop system is stable in the sense of Lyapunov. Moreover, the tracking error is guaranteed to be uniformly asymptotically stable (UAS) rather than uniformly ultimately bounded (UUB) with the aid of an additional robustifying control term. The proposed control algorithm is relatively simple and requires no restrictive conditions on the design constants for the stability. The efficiency of the proposed scheme is shown through the simulation of a simple nonaffine nonlinear system. PMID- 15787149 TI - Soft learning vector quantization and clustering algorithms based on non Euclidean norms: single-norm algorithms. AB - This paper presents the development of soft clustering and learning vector quantization (LVQ) algorithms that rely on a weighted norm to measure the distance between the feature vectors and their prototypes. The development of LVQ and clustering algorithms is based on the minimization of a reformulation function under the constraint that the generalized mean of the norm weights be constant. According to the proposed formulation, the norm weights can be computed from the data in an iterative fashion together with the prototypes. An error analysis provides some guidelines for selecting the parameter involved in the definition of the generalized mean in terms of the feature variances. The algorithms produced from this formulation are easy to implement and they are almost as fast as clustering algorithms relying on the Euclidean norm. An experimental evaluation on four data sets indicates that the proposed algorithms outperform consistently clustering algorithms relying on the Euclidean norm and they are strong competitors to non-Euclidean algorithms which are computationally more demanding. PMID- 15787150 TI - Face membership authentication using SVM classification tree generated by membership-based LLE data partition. AB - This paper presents a new membership authentication method by face classification using a support vector machine (SVM) classification tree, in which the size of membership group and the members in the membership group can be changed dynamically. Unlike our previous SVM ensemble-based method, which performed only one face classification in the whole feature space, the proposed method employed a divide and conquer strategy that first performs a recursive data partition by membership-based locally linear embedding (LLE) data clustering, then does the SVM classification in each partitioned feature subset. Our experimental results show that the proposed SVM tree not only keeps the good properties that the SVM ensemble method has, such as a good authentication accuracy and the robustness to the change of members, but also has a considerable improvement on the stability under the change of membership group size. PMID- 15787151 TI - Multi-aspect target discrimination using hidden Markov models and neural networks. AB - This paper presents a new multi-aspect pattern classification method using hidden Markov models (HMMs). Models are defined for each class, with the probability found by each model determining class membership. Each HMM model is enhanced by the use of a multilayer perception (MLP) network to generate emission probabilities. This hybrid system uses the MLP to find the probability of a state for an unknown pattern and the HMM to model the process underlying the state transitions. A new batch gradient descent-based method is introduced for optimal estimation of the transition and emission probabilities. A prediction method in conjunction with HMM model is also presented that attempts to improve the computation of transition probabilities by using the previous states to predict the next state. This method exploits the correlation information between consecutive aspects. These algorithms are then implemented and benchmarked on a multi-aspect underwater target classification problem using a realistic sonar data set collected in different bottom conditions. PMID- 15787152 TI - Optimizing the kernel in the empirical feature space. AB - In this paper, we present a method of kernel optimization by maximizing a measure of class separability in the empirical feature space, an Euclidean space in which the training data are embedded in such a way that the geometrical structure of the data in the feature space is preserved. Employing a data-dependent kernel, we derive an effective kernel optimization algorithm that maximizes the class separability of the data in the empirical feature space. It is shown that there exists a close relationship between the class separability measure introduced here and the alignment measure defined recently by Cristianini. Extensive simulations are carried out which show that the optimized kernel is more adaptive to the input data, and leads to a substantial, sometimes significant, improvement in the performance of various data classification algorithms. PMID- 15787153 TI - An intelligent adaptive control scheme for postsurgical blood pressure regulation. AB - This paper presents an adaptive modeling and control scheme for drug delivery systems based on a generalized fuzzy neural network (G-FNN). The proposed G-FNN is a novel intelligent modeling tool, which can model unknown nonlinearities of complex drug delivery systems and adapt to changes and uncertainties in these systems online. It offers salient features, such as dynamic fuzzy neural topology, fast online learning ability and adaptability. System approximation formulated by the G-FNN is employed in the adaptive controller design for drug infusion in intensive care environment. In particular, this paper investigates automated regulation of mean arterial pressure (MAP) through intravenous infusion of sodium nitroprusside (SNP), which is one attractive application in automation of drug delivery. Simulation studies demonstrate the capability of the proposed approach in estimating the drug's effect and regulating blood pressure at a prescribed level. PMID- 15787155 TI - Improved access to sequential motifs: a note on the architectural bias of recurrent networks. AB - For many biological sequence problems the available data occupies only sparse regions of the problem space. To use machine learning effectively for the analysis of sparse data we must employ architectures with an appropriate bias. By experimentation we show that the bias of recurrent neural networks--recently analyzed by Tino et al. and Hammer and Tino--offers superior access to motifs (sequential patterns) compared to the, in bioinformatics, standardly used feedforward neural networks. PMID- 15787156 TI - A spatially constrained mixture model for image segmentation. AB - Gaussian mixture models (GMMs) constitute a well-known type of probabilistic neural networks. One of their many successful applications is in image segmentation, where spatially constrained mixture models have been trained using the expectation-maximization (EM) framework. In this letter, we elaborate on this method and propose a new methodology for the M-step of the EM algorithm that is based on a novel constrained optimization formulation. Numerical experiments using simulated images illustrate the superior performance of our method in terms of the attained maximum value of the objective function and segmentation accuracy compared to previous implementations of this approach. PMID- 15787157 TI - An improved conjugate gradient scheme to the solution of least squares SVM. AB - The least square support vector machines (LS-SVM) formulation corresponds to the solution of a linear system of equations. Several approaches to its numerical solutions have been proposed in the literature. In this letter, we propose an improved method to the numerical solution of LS-SVM and show that the problem can be solved using one reduced system of linear equations. Compared with the existing algorithm for LS-SVM, the approach used in this letter is about twice as efficient. Numerical results using the proposed method are provided for comparisons with other existing algorithms. PMID- 15787159 TI - Refractory pulse counting processes in stochastic neural computers. AB - This letter quantitiatively investigates the effect of a temporary refractory period or dead time in the ability of a stochastic Bernoulli processor to record subsequent pulse events, following the arrival of a pulse. These effects can arise in either the input detectors of a stochastic neural network or in subsequent processing. A transient period is observed, which increases with both the dead time and the Bernoulli probability of the dead-time free system, during which the system reaches equilibrium. Unless the Bernoulli probability is small compared to the inverse of the dead time, the mean and variance of the pulse count distributions are both appreciably reduced. PMID- 15787160 TI - Genetic alterations in B-cell non-Hodgkin's lymphoma. AB - BACKGROUND: Although the patients with diagnosed B-NHL are classified into the same disease stage on the basis of clinical, histopathological, and immunological parameters, they respond significantly different to the applied treatment. This points out the possibility that within the same group of lymphoma there are different diseases at molecular level. For that reason many studies deal with the detection of gene alterations in lymphomas to provide a better framework for diagnosis and treatment of these hematological malignancies. AIM: To define genetic alterations in the B-NHL with highest possibilites for diagnostic purposes and molecular detection of MRD. METHODS: Formalin fixed and paraffin embedded lymph node tissues from 45 patients were examined by different PCR techniques for the presence of IgH and TCR gamma gene rearrangement; K-ras and H ras mutations; c-myc amplification and bcl-2 translocation. There were 34 cases of B-cell non-Hodgkin's lymphoma (B-NHL), 5 cases of T-cell non-Hodgkin's lymphoma (T-NHL) and 6 cases of chronic lymphadenitis (CL). The mononuclear cell fraction of the peripheral blood of 12 patients with B-NHL was analyzed for the presence of monoclonality at the time of diagnosis and in 3 to 6 months time intervals after an autologous bone marrow transplantation (BMT). RESULTS: The monoclonality of B-lymphocytes, as evidenced by DNA fragment length homogeneity, was detected in 88 % (30/34) of B-NHL, but never in CL, T-NHL, or in normal PBL. Bcl-2 translocation was detected in 7/31 (22.6%) B-NHL specimens, c-myc amplification 9/31 (29%, all were more than doubled), K-ras mutations in 1/31 (3.23%) and H-ras mutations in 2/31 (6.45%) of the examined B-NHL samples. In the case of LC and normal PBL, however, these gene alterations were not detected. All the patients (12) with B-NHL had dominant clone of B-lymphocyte in the peripheral blood at the time of diagnosis while only in 2 of 12 patients MRD was detected 3 or 6 months after BMT. CONCLUSION: Because it is quic and simple, PCR analysis of clonal IgH rearrangements is very useful when diagnostic assistance is required. This technique is also very effecient for tracking minimal residual disease in lymphomas and leukemias and for monitoring clonal evolution in acute and chronic lymphoblastic leukemias and lymphomas. The presence of other genetic alterations, which we detected, should serve as an additional prognostic or predictive factor in the patients with B-NHL. PMID- 15787161 TI - Arrhythmias and conduction abnormalities in children after repair of tetralogy of Fallot. AB - AIM: To find out types and frequency of cardiac arrhythmias and conduction abnormalities in the group of children who underwent surgery for tetralogy of Fallot (TOF). METHODS: Forty-six pedicatric patients who underwent a complete repair of TOF at the age of 1 to 13 (mean 2.89+/-2.36) were studied Thirty-eight (82.60%) had total correction and 8 (17.40%) had palliative operation first, and total correction afterwards. Twenty-four-hour Holter ECG monitoring was performed in all 46 pediatric patients aged from 1 to 16 yrs (mean 6.48+/-4.04) after surgery as follows: in 1 patient (2.17%) after a year, in 20 patients (43.477%) after 2 to 5 years and in 25 patients (54.34%) after 5 years. Mean age of patients on Holter monitoring was 9.25+/-4.39 (range 4-19). Twenty of them (43.47%) were girls and 28 (56.53%) were boys. All the patients were evaluated by standard methods (clinical signs, clinical findings, ECG before surgery, ECG before Holter monitoring and 2D Doppler echocardiography. RESULTS: Types of heart rhythm found out by Holter monitoring were: sinus nodus dysfunction in 1 child (2.17%), significant premature atrial contraction (PAC) in 8 (17.39%), supraventricular paroxysmal tachycardia in 3 (6.535), transient nodal rhythm in 2 (4.34%), premature ventricular contraction (PVC) Lown grade 1-III in 9 (19.56%) and Lown grade IV in 2 (4.34), atrioventricular (AV) block grade I in 2, right bundle branch block (RBBB) in all 46 (100%) and RBBB + left anterior hemiblock (LAH) in 4 (8.96%). There was no presence of atrial flutter, ventricular tachycardia or complete A V block. None of them experienced sudden death. Using cross procedure statistical methods, it was found that all the patients with PVC had right ventricular dilatation. There was no relation of other types of arrhythmia found on Holter monitoring to the other parameters from echocardiography, neither to the other standard methods. Children did not need the pace-maker, but 36.95% of the them required antiarrhythmic drugs. CONCLUSION: Twenty-four hour Holter ECG is a noninvasive and very sensitive method for discovering heart rhythm disturbances in children after the repair of tetralogy of Fallot, especially in asymptomatic patients. The patients after the repair of this congenital heart disease needed a long-term follow-up for early recognition of serious heart rhythm disturbances and their treatment. PMID- 15787162 TI - [Significance of transvaginal Doppler ultrasonography for detection of malignant gestational trophoblastic disease]. AB - Transvaginal Doppler ultrasonography has an important role in detecting and confirming the gestational trophoblastic disease (GTD). It can also be helpful in early detection of the malignant cases of GTD such as the invasion and protrusion of trophoblastic tissue into the uterine wall. Ultrasonographic picture of the malignant GTD is also specific for the presence of prominent zones of vasculavization in the peritrophoblastic tissue, as well as in the uterine tissue in which malignant GTD is developed. Resistance Index (RI) values were measured at the level of blood vessels of peritrophoblastic tissue and of suspected zones in the uterine tissue to detect neovascularization, which followed the malignant process. Theca luteal cysts were often detected by Transvaginal Doppler ultrasonography. PMID- 15787163 TI - Evaluation of absolute volume of human fetal kidney's cortex and medulla during gestation. AB - BACKGROUND: Human fetal kidney is quite different from the mature kidney, both macroscopically and hystologically. Lobulated surface of the human fetal kidney reflects its inner organisation. AIM: To determine the fetal kidneys' volume according to the gestational age, to establish periods of their maximal and minimal growth and to compare these values for various gestational ages. METHODS: Forty five human fetal kidneys aged from IV to X lunar months were analyzed. Kidneys were divided into nine groups according to their gestational age. The volumes of cortex and medulla were determined using stereological methods. The results were statistically analyzed and the periods of significant growth of these structures were marked. RESULTS: Fetal kidney's cortex and medulla grew continually with a very high coefficient of linear correlation with crown-rump length. The cortex/medulla ratio was minimal in the first half of V lunar month, when medulla grew most rapidly and it was maximal immediately before birth, when cortex achieved its maximum. CONCLUSION: This study was an effort to provide some parameters which would help in the future investigations of the development of human fetal kidney. PMID- 15787164 TI - [Predictive factors of psychosomatic reactions during air raids]. AB - BACKGROUND: Civilian population of Yugoslavia was exposed to massive stressors of air raids by NATO in 1999. The aim of this study was to investigate somatic complaints and their predictive factors during stresses of air raids. METHODS: Random sample of 434 subjects, consisting of 139 inhabitants of several Belgrade municipalities and 295 employees of Military Medical Academy, were assessed in the cross-sectional study. The basic factors of interest were stress severity, variables of personality and habits and behaviour relevant for somatic complaints. Self-report of stress severity and the most common somatic complaints were performed by Questionnaire specially designed for this purpose and personality evaluation by EPQ-38. Multiple regression analysis was used to determine the influence of predictors on dependent variable (psychosomatic symptoms). RESULTS: Personality was the most important predictor that explained 29% of variance in somatic symptoms (with the strongest impact of neuroticism); 11% was explained by habits and behaviour and only 1% of variance by stress. CONCLUSION: The finding that personality had higher impact on stress reaction outcome could be important for preventive and therapeutic aspects of stress reactions. PMID- 15787165 TI - [Effects of adenoidectomy and immediate orthodonthic treatment on jaw relations and naso-respiratory rehabilitation]. AB - AIM: To determine if adenoidectomy itself could provide spontaneous relation of the transverse growth of jaws as well as an adequate mode of breathing or if there is a need for an additional orthodontic treatment to solve the problem. METHODS: The study included one hundred and one patients aged 6-17 yrs., divided into three groups: group K--patients with oral respiration caused by adenoidal enlargement; group 1--patients with adenoidectomy done 5 or more years before; group 2--patients with orthodontic treatment done immediately after adenoidectomy. Transverse occlusion relation, the frequency of crossbite, and the intensity of nasal respiration were measured in all three groups of patients. RESULTS: The obtained results have shown that in the patients with adenoidal enlargement and oral respiration, there was an evidence of maxillary width reduction as opposed to mandibular width. Of the patients, 14% were with unilateral or bilateral crossbite. In the patients with adenoidectomy done 5 or more years before, transverse maxillary growth compared to mandibular one in the region of the front width, showed statistically significant increase (p < 0.05) as opposed to the patients with adenoidal enlargement. The effect of adenoidectomy on transverse maxillary growth was not satisfactory anyway, which was also indicated by the increase of frequency of crossbite in 24% of the patients with adenoidectomy done 5 or more years before. In operated on and in the patients one month after orthodontic treatment, transverse maxillary growth compared to mandibular growth in all the regions of both front and back width, there was statistically highly significant increase (p < 0.01) as opposed to the patients with adenoidal enlargement and adenoidectomy done many years before. Significant maxillary and mandibular transverse relation was obtained without the presence of crossbite. Nasal respiration prevailled. CONCLUSION: Orthodontic therapy one month after adenoidectomy was necessary for solving the orthodontic problems caused by adenoidal enlargement and significantly contributed to the rehabilitation of nasal respiration. PMID- 15787166 TI - [Morphometric analysis of neurons from the marginal and substantia gelatinosa layers of human spinal cord: classification according to laminar organization]. AB - The main goal of morphometric analysis of neuronal images, except for getting information about their geometry and dendritic branching patterns, is their classification based on laminar organization. The majority of contemporary techniques for image analysis are based on the application of fractal theory, which has some limitations on results analysis. For that reasons, the new, mostly nonfractal techniques for image analysis had been designed in the past few years. This study shows the analysis of morphometry of the human spinal cord neurons from the marginal (lamina I) and substantia gelatinosa (laminae I-II). For the analysis of neuron images two techniques of morphometric analysis were used: box counting method as a mainly used technique for fractal analysis, and circle counting method as a newly designed technique for measuring the length of dendrites. The use of these methods for neurons of the mentioned regions of human spinal cord showed that circle-counting method had given more accurate results than fractal analysis method. When the proposed method was used for the analysis of neuronal images, it was possible to classify neurons according to their laminar position. PMID- 15787167 TI - [Thymic nurse cells--a specialized thymic microenvironment]. PMID- 15787168 TI - [Natriuretic peptides in clinical practice]. PMID- 15787169 TI - [Intraorbital wooden foreign body, diagnosed by NMR and ultrasonography, and extirpation by cranio-orbital approach]. AB - AIM: The case of the phlegmon of the left orbit associated with ophtalmoplegia and blindness in a 16-years old boy was presented. It was caused by the penetrafion of the wooden foreign body under unknown circumstances. CASE REPORT: Repeated CT scans did not reveal the presence of the foreign body before MR imaging and ultrasonography. The removal (43 x 8 mm) was done one month after the injury by osteoplastic orbitotomy, using microsurgical technique. Three months later the signs of inflammation and eye protrusion withdrew. Motility of the left upper lid and eye were normal. CONCLUSION: MR imaging and ultrasonography were decisive in the diagnose of organic foreign body in the orbit. PMID- 15787170 TI - [Significance of iron reduction for the therapy of chronic hepatitis C]. AB - BACKGROUND: It has been established that many patients with chronic hepatitis C have elevated serum iron, feritin levels and iron deposites in the liver. Therefore, the liver damage due to hepatitis C virus may be aggravated with iron overload. In many studies higher levels of iron in the blood and the liver were connected with the decreased response to interferon-alfa therapy for chronic viral hepatitis C. Recent introduction of pegylated interferons plus ribavirin has improved the therapeutic response, so it is now possible to cure more than 50% of the patients. CASE REPORT: Three patients with chronic hepatitis C and iron overload were presented. Iron reduction therapy using phlebotomy or eritrocytapheresis with plasmapheresis was done at different times in regard to specific antiviral therapy or as a sole therapy. CONCLUSION: It has been shown that iron reduction, sole or combined with antiviral therapy, led to the deacreased aminotransferase serum activity and might have slown down the evolution of chronic hepatitis C viral infection. PMID- 15787171 TI - [Stunned myocardium]. AB - BACKGROUND: Stunned myocardium is a state of delayed recovery of regional contractility after a transient period of ischemia followed by reperfusion. CASE REPORT: A 67-year-old patient was admitted to our hospital with acute anterior myocardial infarction, and treated using percutaneous transluminal coronary angioplasty (PTCA) within acute disease stage. Reversible myocardial dysfunction persisted after ischemia following the return of normal perfusion. Abnormal resting wall motion with augmentation of contractility at low and high doses of dobutamine characterizes the stunned myocardium and reflects the normal blood flow reserve, characteristic for these postischemic, reperfused segments. SPECT (Single Photon Emission Computerized Tomography) with Tc 99 and dipyradamole showed normalization of perfusion defects in the apical region. There months after the infarction and PTCA, contractility was almost completely recovered. CONCLUSION: Stunned myocardium recovery lasted from few weeks to few months. Control ultrasonography as well as SPECT showed normalization of systolic function of the left ventricle in the viable segments registered at previous examinations. PMID- 15787172 TI - [Culture of the corneal epithelium--comparison of the mitotic potential of limbal cells from living and cadaveric donors]. AB - PURPOSE: We analyse effectiveness of corneal limbal cells' culture prepared from heart-beating organ donors, that include living donors and from cadaver donors buttons following 3 days storage in 4 degrees C in Likorol. MATERIAL AND METHODS: For experiment 12 adults (living and heart-beating organ donors) aged 28-63 (mean 46.3) and 12 corneal buttons of cadaver (aged 18-51, mean 34.1) were qualified. Tissue samples (1 mm2) were taken from superior corneal limbus. Sample from living donor obtained during routine operation was sent immediately to laboratory, as well as from heart-beating organ donor. The limbal biopsy of preserved cornea was taken after 3 days storage in 4 degrees C (Likorol). Samples were treated with trypsin/EDTA solution before culture. Collected cells in similar density in 1 ml of medium laid on dishes inserts, covered with fibrin (Tissucol) and cultured in presence of feeder layer of fibroblasts (L929 line). Epithelial cells were cultured for 14 days at 37 degrees C in humidified 5% CO2 atmosphere in supplemented 2:1 mixture of DMEM and Ham's F12. On the 14th day cells were collected. Number of cells per 1 ml of medium was counted by cytometer. Immunostaining for epithelium type (Keratin 3) was performed. RESULTS: The number of cells obtained from cadaver donors reached 184.2+/-14.9% whereas from living donors revealed 1013.1+/-104.2%, increase in relation to number of delivered cells. We observed only 0.83+/-0.3 colonies per microscopic area in cultures from preserved tissue versus 6.67+/-0.6 colonies in cultures from living donor. CONCLUSIONS: The preservation in 4 degrees C in Likorol significantly decreases proliferative potential of the corneal limbus. PMID- 15787173 TI - [Genetic analysis of rhodopsin and peripherin genes in patients with autosomal dominant retinitis pigmentosa (adRP) in Polish families]. AB - PURPOSE: The aim of that study was to identify the mutations in rhodopsin and peripherin genes in Polish families with autosomal dominant form of retinitis pigmentosa and determine the population polymorphism in both genes in adRP families. MATERIAL AND METHODS: We performed ERG, visual acuity, Goldman visual fields, intraocular pressure measurements and fundoscopy in all the patients included in the study. On the basis of disease history, the families pedigree was made and the mode of inheritance was analyzed. The molecular analysis of DNA for each family with adRP was conducted. Genomic DNA was obtained from leucocytes by phenol-chloroform procedure according to Maniatis protocol. DNA was amplified by the PCR reaction in a volume of 50 microl containing 100 ng/microl of genomic DNA, water, Cetus buffer pH 8.4 (1 n Tris, 1 n MgCl, 1 n KCl, 2% gelatin), 0.25 microM of each primer, 200 microM of each of dATP, dTTP, dCTP, and dGTP and 2.5 U Taq polymerase (Promega). For amplification of rhodopsin gene 30 cycles of PCR were carried out. Each cycle consisting of denaturation at 95 degrees C for 5 min, annealing: at 58 degrees C (exon 1), 63 degrees C (exon 2 and 3), 68 degrees C (exon 4) and 2 min extension at 72 degrees C min. For amplification of peripherin gene 30 cycles of PCR were carried out with annealing at 60 degrees C. The entire PCR product was in electrophoresis on 8% PAA. The PCR-RFLP PCR-HD PCR SSCP and analysis of polymorphism (CA)n dinucleotide repetition was performed. RESULTS: Molecular study demonstrated, that mutations in rhodopsin gene were cause of retinitis pigmentosa in case of two families. In any study families mutations in peripherin gene were not identified. Two kinds of bases polymorphism were identified: restriction fragments length polymorphism (RFLP) in rhodopsin gene in exon 1 and 3 and single strand conformation polymorphism (SSCP) in exon 1 and 3 in rhodopsin gene and in exon 3 in peripherin gene. The confirmed mutations in rhodopsin gene, cosegregation with adRP, whereas two kinds of population polymorphism did not correlate with clinical symptoms. Natural polymorphism appeared to be a frequent feature in rhodopsin gene while a less frequent feature in peripherin gene. CONCLUSIONS: Genetic investigations in patients with adRP allow to confirm the diagnosis and evaluate the prognosis. The mutation in rhodopsin gene should be confirmed in directly sequencing reaction in next study. PMID- 15787174 TI - [The influence of viscoelastic's maintenance in the anterior chamber on early results and complications after trabeculectomy]. AB - PURPOSE: The evaluation of intraocular pressure and number of complications in the early postoperative period in patients after trabeculectomy with viscoelastic agent used to recreate the anterior chamber at the end of procedure. MATERIAL AND METHODS: Between November 2001 and February 2002, in 56 patients (58 eyes) with primary open-angle glaucoma trabeculectomy was performed. Patients were divided into two groups. Trabeculectomy was performed with leaving the viscoelastic agent (Provisc) in the anterior chamber at the end of surgery in 29 eyes of 28 patients (Group A), while in 28 patients (29 eyes) the anterior chamber was filled with Ringer solution (Group B). Preoperative IOP in group A ranged from 19 to 48 mmHg, mean--28.41 mmHg. In group B it ranged between 19 and 41 mmHg, mean--25.07 mmHg. The follow-up time was 1 month in every case. RESULTS: The mean IOP on the first, fifth, 14th and 30th day after surgery was 14.0 mmHg, 13.2 mmHg, 12.03 mmHg and 11.76 mmHg respectively in group A, while in group B corresponding values were: 9.0 mmHg, 8.97 mmHg, 8.83 mmHg and 8.9 mmHg. The shallowing of anterior chamber and choroidal detachment were observed in 4 eyes (13.8%) in group A and in 8 eyes (27.6%) in group B postoperatively. The increased IOP during first days after surgery was observed in 3 cases (10.3%) in group A. CONCLUSIONS: The viscoelastic agent left in the anterior chamber at the end of trabeculectomy significantly decreases the frequency of anterior chamber shallowing and choroidal detachment and reduces number of cases with hypotony. PMID- 15787176 TI - [Surgically induced astigmatism after cataract phacoemulsification]. AB - PURPOSE: To evaluate the astigmatic effect after phacoemulsification depending on size and location of incision. MATERIAL AND METHODS: Three groups of 30 patients each: In 1st group with scleral tunnel incision enlarged to 6 mm with continuous cross-like suture at 12 o'clock; In 2nd group with 3.2 mm scleral incision without suture at 12 o'clock; In 3rd group with 3.2 mm superotemporal incision in clear cornea, no suture. Postoperative astigmatism was examined by keratometry after 1 day, 1 week and 1 and 3 months post surgery. Surgically induced astigmatism was calculated by polar values method. RESULTS: After 3 months observation the lowest mean postoperative corneal astigmatism was achieved in group III. The SIA was 0.71 in group III, 1.08 in group I and 0.95 in group II. The differences between group III and groups I, II were statistically significant. CONCLUSIONS: Surgically induced astigmatism can be minimized with incision in clear cornea and no suture. PMID- 15787175 TI - [The estimation of efficacy of the phacotrabeculectomy procedure in patient with cataract and pseudoexfoliation glaucoma]. AB - PURPOSE: The aim of the study was the estimation of phacotrabeculectomy effectiveness in patient with pseudoexfoliation syndrome. MATERIAL AND METHODS: The analyzed group consisted of 72 eyes of 65 patients in the range of 61-92 years old. 32 patients have phacoemulsification procedure (gr. I) and 40 patients have performed the procedure of the phacotrabeculectomy (gr. II). RESULTS: In 60% of the eyes marked postoperative improvement of visual acuity to more than 0.6 was observed. We noticed significant decrease of mean intraocular pressure (from 22.45 mmHg-17.88 mmHg), only in group II. CONCLUSIONS: We have concluded that the polyprocedular operation of phacotrabeculectomy in cataract with pseudoexfolation syndrome is an efficient procedure lowering mean intraocular pressure and improving visual acuity. PMID- 15787177 TI - [Refractive changes in children with pseudophakia]. AB - PURPOSE: To report refractive changes after cataract surgery and intraocular lens implantation in children. MATERIAL AND METHODS: 104 patients (123 eyes) operated between January 1987 and December 1997 with congenital or traumatic cataract. Refractive power in both operated and non operated eyes, were measured. RESULTS: The operated eyes tended to become myopic. Children, who underwent surgery in the first 6 years of life had a greater myopic shift than older children. The mean myopic shift ranged from -2.33 D to -0.51 D in operated eyes and from -1.05 D to 0.49 D in non operated eyes. CONCLUSIONS: Pseudophakia in children is predicted to result in a large quantity of myopic shift, particularly in very young children. This natural myopic shift is magnified by the optical effect of a high power IOL. PMID- 15787178 TI - [Astigmatism at post operational stage after a phacoemulsification cataract removal]. AB - OBJECTIVE: Assessment of post operation astigmatism and dynamics of its reduction within three month after the procedure. MATERIAL AND METHODOLOGY: The research included cataract removal in 140 eyes by means of phacoemulsification (100) and extracapsular (40) methods. Central keratotomy and refraction measurements were conducted with a refractor on 2nd, 7th, 30th and 90th day after the procedure. RESULTS: Average value of postoperative astigmatism after an extracapsular cataract removal was 4.69 D on the second day. After a phacoemulsification without a suture the value was 1.36 D, with a single loop suture -1.98 D, with a single criss-cross suture it was 2.89 D. Two alternative parameters of astigmatism reduction were suggested: a half-value period and a rate of reduction. CONCLUSIONS: Average early postoperative astigmatism values after an extracapsular cataract removal are at least twice as high as in the case of a phacoemulsification cataract removal. A suture applied after a phacoemulsification results in an increase in early postoperative astigmatism. Astigmatism half-value period and rate of reduction provide a good description of the dynamics of postoperative astigmatism reduction. PMID- 15787179 TI - [Simultaneous phacoemulsification, posterior chamber lens implantation and pars plana vitrectomy--clinical observation]. AB - PURPOSE: The aim of the study was to present our experience and observations with phacovitrectomy and IOL implantation. MATERIAL AND METHODS: 60 patients, aged 26 74 years (mean 52 years), 36 women, 24 men: 5 (8.3%) with idiopathic macular hole /I group/, 5 (8.3%) macular pucker /II group/, 39 (65%) with rhematogenous retinal detachment /III group/, 7 (11.7%) with proliferative diabetic retinopathy /IV group/, and 4 (6.6%) with vitreous hemorrhage /V group/. RESULTS: Visual acuity improved in 44 (73.3%) patients, remained unchanged in 7 (11.6%) and was worse in 9 (15%) cases. The most common intraoperative complication was oedema of the corneal epithelium in 13 (21.7%) cases and postoperational raise of the intraocular pressure in 13 (21.6%) cases. CONCLUSIONS: Phacovitrectomy does not significantly raise up the number of complications, it is safe and beneficial for both, the patient and the surgeon. It must be born in mind that post operational fibrin formation after phacovitrectomy is bigger. PMID- 15787180 TI - [Radial optic neurotomy (RON) in central retinal vein occlusion (CRVO)- preliminary report]. AB - PURPOSE: Practical application of radial optic neurotomy for patients with central retinal vein occlusion, based on our initial experience. MATERIAL AND METHODS: Three patients with central retinal vein occlusion underwent pars plana vitrectomy with radial optic neurotomy at the nasal site of the optic disc. RESULTS: Improvement of visual acuity by one or three lines in two of three patients who underwent RON. Partial clearing of retinal haemorrhages and retinal oedema occurred in all cases. No serious complications developed intra- and postoperatively. CONCLUSIONS: Radial optic neurotomy remains a controversial but also promising treatment for eyes with CRVO. Further study is necessary to establish RON as a standard therapy for CRVO. PMID- 15787181 TI - [Evaluation of effectiveness of Catalin eyedrops in patients with presenile and senile cataract]. AB - AIM OF THE STUDY: Evaluation of effectiveness of Catalin eye drops in inhibition of cataract or eventual influence on regression of actually present opacities. PATIENTS AND METHOD: 72 patients in age over 40 years with initial cortical cataract and visual acuity better than 0.5 were evaluated. 35 patients were treated with anticataracous drug Catalin, and 37 received placebo. All patients were evaluated with a slit-lamp and modern equipment EAS-1000 (Nidek) before starting research and after 3, 6, 12, 18 and 24 months of treatment. RESULTS: Catalin-eye drops in patients with cortical cataract showed its effectiveness in inhibition of lens opacification and its progression especially in group of patients in age up to 59 years. The increase of lens opacification in this group was only 1.425% (Catalin), and in older group (placebo) even 9.228%. Appropriate, raising effect of the drug can be observed after 18 months of treatment. Catalin eye drops did not cause any complications during the study. CONCLUSIONS: 1. Catalin-eye drops used in patients with senile and presenile cortical cataract is effective drug in inhibiting lens opacification. 2. The function of the drug is more obvious in group of younger patients (up to 59 years of age) in comparison with older ones. 3. Appropriate, raising effect of the drug can be observed after 18 months of treatment. 4. Catalin-eye drops is well tolerated by the patients even after prolonged application. PMID- 15787182 TI - [Ocular complications in leukemias and malignant lymphomas in children]. AB - PURPOSE: The aim of the study was to determine the prevalence and types of ocular abnormalities in children with leukemias or lymphomas. MATERIAL AND METHODS: We have studied 39 children (age 0.5-17 years) with leukemia or lymphoma. Ocular manifestations were present in 54% of study group (21 children). The most frequent ocular findings were seen in conjunctiva (33.4%). 15.4% patients presented with posterior segment findings, without loss of ocular acuity. Other symptoms were dry eye syndrome and proptosis. CONCLUSIONS: Children treated for leukemia or lymphoma should be systematically examined by ophthalmologist. Ophthalmic complications can be results of basic disease, treatment or manifestations of relapse. PMID- 15787183 TI - [Acute endophthalmitis of 15-year old boy in the course of acute lymphoblastic leukemia--part I]. AB - PURPOSE: The aim of this study is to present dramatic ocular manifestation of acute lymphoblastic leukemia. MATERIAL AND METHODS: 15-year old boy was hospitalized due to acute leukemia at the Department and Clinic of Pediatric Hematology and was referred to ophthalmologist because of strong pain and decreased visual acuity of the left eye. General ophthalmic examination and electrophysiology were done. These procedures were repeated after 1, 2, 5 and 11 weeks and also after 6 and 9 months. RESULTS: Acute uveitis with cellular exudation to the vitreous, papilledema, swelling of the macula and green yellowish infiltration which elevated the temporal part of the retina, were noticed. Swelling of the iris and miosis were observed. After one week of local anti-inflammatory treatment the eye became painless with little deep injection only. Simultaneously causal treatment of basic disease was continued. Control examinations revealed step by step improvement of ocular changes. After 6 months in the place of retinal infiltration choroidoretinal atrophy was seen. The functional deficits in visual field and electrophysiological examinations were found, too. One year later, the patient came again because of recurrence. Involvement of the central nervous system with signs of meningitis occurred. Visual acuity was normal and no infiltration of eyes was found, but there was papilledema in both eyes. CONCLUSIONS: In acute leukemia ocular manifestation may be highly expressed. The patients require local symptomatic and general causal treatment in cooperation of ophthalmologist with hematologist. PMID- 15787184 TI - [Acute endophthalmitis of 15-year old boy in the course of acute lymphoblastic leukemia. Part II--Electrophysiological examinations]. AB - PURPOSE: The aim of the study is to present objective estimation of visual function in eyes affected by leukemia. MATERIAL AND METHODS: The boy who was described in the first part of this study was examined. Visual evoked potential (VEP), full-field flash electroretinography (FERG), electrooculography (EOG), were done according to ISCEV standards. RESULTS: The function of retina was decreased proportionally to the areas of infiltration. The amplitude of VEP peaks was lowered when the involvement of the central nervous system occurred. In the eye with retinal infiltration abnormal EOG was present a few months earlier than pigment epithelium destruction was seen in ocular fundus. CONCLUSIONS: Functional deficits may be irreversible. PMID- 15787185 TI - [Simultaneous phacoemulsification, posterior chamber lens implantation and pars plana vitrectomy]. AB - The aim of the study was to present current knowledge in the field of combined surgery: phacoemulsification with intraocular lens implantation and pars plana vitrectomy. PMID- 15787186 TI - [Recent developments in kinetic perimetry]. AB - The manual kinetic perimetry using Goldmann instrument is still widely used and remains a "gold standard" in examining patients with any kind of advanced visual field loss and impaired co-operability. Nevertheless, Goldmann instrument has several disadvantages, such as examiner dependence, response variability, lack of standardization of parameters, poor repeatability. The latest development in kinetic perimetry, which was realized at the University Eye Hospital in Tuebingen (Germany) is semi-automated kinetic perimetry (SKP), using Octopus 101 instrument (Interzeag/Haag-Streit, Koeniz-Bern, Switzerland). It is an examiner-independent, standardized and reproducible method. Additionally, obtained results can be corrected for the mean individual reaction time. This paper will briefly present characteristic, advantages and application of SKP in clinical practice. PMID- 15787187 TI - [Macular corneal dystrophy--clinical state, histopathologic, immunohistochemical examinations and genetical dependence]. AB - Macular corneal dystrophy is a rare corneal disease, with autosomal recessive inheritance, in which characteristic diffuse corneal clouding and reduction of corneal thickness are present. Authors in this article presented the clinical state and molecular pathology underlying it's importance in diagnosis of the macular corneal dystrophy. As in other inherited diseases, the latest achievement of molecular biology, concerning the new mutation of the CHST6 gene, changed the classification of the macular corneal dystrophy, and can have the significant influence on genetic therapy of this disease. PMID- 15787188 TI - [Changes in organ of vision in proliferative diseases of hematopoietic system in children]. AB - Leukemias and lymphomas are the most common neoplasms of hematopoietic system in children. The aim of the study was to determine ophthalmological signs in these disaeses referring to available literature. PMID- 15787190 TI - [Report from the congress of American Academy of Ophthalmology in New Orleans]. PMID- 15787189 TI - [The legal evaluation of contacts between a physician and a pharmacological company]. PMID- 15787191 TI - New approaches to development of mucosal vaccine against enteric bacterial pathogens; preventing campylobacteriosis. AB - Although vaccination, after having been more than 200 years in medical practice, has proven to be the most effective and the cheapest way to prevent infectious diseases, they remain still the main cause of human premature deaths. As many pathogens enter the human body through the mucosal surfaces, the mucosal way of immunization is considered to be the most promising strategy to decrease the number of human infections. Moreover, the oral delivery system eliminates the necessity of injection what is extremely important for pediatric immunization programs. However, most of recently constructed subunit vaccines based on purified bacterial/viral antigens are rather poorly immunogenic. This review presents some novel ways to enhance and modulate host immune responses by combining antigens with specific adjuvants or by employing specific delivery systems. We also discuss some recent technologies, based on mining the genomic sequences of bacterial pathogens, which accelarate and improve identification of new candidates for vaccine construction. As an example, we focus on the progress in the development of vaccine against Campylobacter spp. Campylobacter jejuni is now recognized as a leading cause of bacterial enteritis in human. PMID- 15787192 TI - Molecular aspects of Listeria monocytogenes infection. AB - Listeria monocytogenes, a food-borne intracellular animal and human pathogen, interacts with infected host cells both prior to entry and during the intracellular phase of infection. This review is focused on the role of secreted proteins, including listeriolysin O and two distinct phospholipases C, in modulating the signal transduction of infected cells. PMID- 15787193 TI - Optical lectin based biosensor as tool for bacteria identification. AB - Biosensor techniques are based on biospecific interaction between the biological parts of biosensor with the analyte. In biosensor construction, antibodies are usually used for the detection of analytes such as microorganism, because of very strong and highly specific interaction. The disadvantages of this assay are a long time needed for antibody isolation and purification as well as difficult regeneration of biosensor chip. The use of lectins instead of antibodies could solve these problems because a several hundred lectins are commercially available and their stability in standard buffers is better compared to monoclonal antibodies. While antibody can only be used to detect that antigen it was designed for, lectin as low affinity molecule may bind several different pathogens. Using the discriminative effect of an artificial neural network the application of a lectin array will compensate for the lower specificity. Microbial surfaces bear many of the sugar residues capable of interacting with lectins. The ability of lectins to react with microbial glycoconjugates means that it is possible to employ them as probes and sorbents for whole cells, mutants and numerous cellular constituents and metabolites, and it makes them useful tools for identification or typing of bacteria. Lectins are attractive reagents for the clinical diagnostic laboratory because of their diverse specificity, commercial availability, a wide range of molecular weights, and their stability in standard buffers. The construction of lectin biosensor could be an advantage method for detection of pathogenic bacteria. PMID- 15787194 TI - Analysis of the peptidoglycan hydrolases of Listeria monocytogenes: multiple enzymes with multiple functions. AB - Listeria monocytogenes is a ubiquitous gram-positive, rod-shaped, widespread in nature, facultative intracellular human and animal pathogen that causes infections collectively termed listeriosis. L. monocytogenes EGD encodes a total of 133 surface proteins, the abundance of which, as well as the variety of anchoring systems, probably reflects the ability of this bacterium to survive in diverse environments and to interact with many kinds of eukaryotic cells. The group of surface proteins also includes proteins with murein hydrolase activity autolysins. To date, five L. monocytogenes autolysins have been identified: p60, P45, Ami, MurA and Auto. These enzymes are involved in numerous cellular processes including cell growth, cell wall turnover, peptidoglycan maturation, cell division and separation, formation of flagella, sporulation, chemotaxis and biofilm formation, genetic competence, protein secretion, the lytic action of some antibiotics and pathogenicity. We have recently identified a putative sixth listerial peptidoglycan-degrading enzyme, which has surprisingly been identified as FlaA, a flagellar protein of L. monocytogenes. PMID- 15787195 TI - Polymerizer-mediated intracellular movement. AB - Bacterial movement inside the cytoplasm is a major virulence factor in that it is necessary for efficient colonization of the infected tissues. Molecules from both the host and the pathogen present possible sites of pharmacologic intervention. Because locomoting Listeria and Shigella mimic the activated state of the leading edge of nonmuscle cells, these pathogens are powerful tools for dissecting the molecular machinery of actin-based motility. Analysis of the movement linked to cytoskeleton may lead to: (I) improved understanding of the mechanisms of disease transmission, including carriers and carrier states, pathogen movements, environmental factors and pharmacokinetics of the uptake and residues of vaccines and other biologics, and drugs in cultivated organisms; (II) new therapeutic developments, since it identifies the molecular targets involved in the pathogenicity of Listeria and Shigella and vaccinia intracellular enveloped virus. Recent knowledge about the intracellular movement in cytoplasm may lead to a better understanding of the processes governing actin dynamics within the cell and disease spread. PMID- 15787196 TI - New methods of pathogenic bacteria elimination. AB - The growing bacterial resistance to antibiotics calls for the elaboration of new pathogens elimination strategies. Some of these methods are based on the conjugative transfer of recombinant plasmids able to eliminate pathogenic recipients by plasmid run-away replication or by killing activity of plasmid encoded bacteriocins. Using live bacteria as donors of plasmid vectors carrying killing determinants requires meeting many safety restrictions in order to eliminate potential biohazard. PMID- 15787197 TI - Molecular modifications of host cells by Toxoplasma gondii. AB - Toxoplasma gondii, the etiological agent of toxoplasmosis, is an Apicomplexa obligate intracellular protozoan parasite, which is able to infect any nucleated cell of numerous endothermic vertebrates. The combined abilities to actively penetrate host cells and perfectly control the fate of the parasite-containing vacuole (parasitophorus vacuole, PV) contribute to the remarkable global success of Toxoplasma as an intracellular parasite. Very broad host range and the relative ease of growth both in cell cultures in vitro and in vivo suggest that the parasite is able to manipulate the host cell apoptotic machinery. The article describes different aspects of host-parasite interplay focusing on molecular modifications of infected host cells. PMID- 15787198 TI - Babesia sp.: emerging intracellular parasites in Europe. AB - The emergence of Lyme borreliosis as the most prevalent arthropod disease of humans in the temperate northern hemisphere has resulted in renewed interest in human babesiosis, transmitted by the same tick vectors. The advent of new molecular tools has made possible a reappraisal of the main parasites involved (Babesia divergens in Europe and Babesia microti in the USA). B. divergens is probably restricted to European cattle, though there are several nearly identical species. B. microti occurs as a world-wide species complex rather than as a single species, and although both phenotypic and genotypic features lend support to suggestions that zoonotic B. microti may occur in Europe, convincing medical evidence is lacking. Comparative biology should support genetic data in taxonomic studies of these parasites. PMID- 15787199 TI - Zoonotic reservoir of Babesia microti in Poland. AB - Babesiosis is as one of the emerging human and animal diseases transmitted by ticks. It is caused intraerythrocytic parasites of the genus Babesia. Current evidence of human babesiosis suggests that the majority of cases are involved by Babesia divergens and Babesia microti piroplasms. As zoonotic reservoir of B. microti serve small mammals--insectivores and rodents. The occurrence of this parasite in natural environment in Poland is documented for various regions, in the wide range of mammal hosts. The most important role as Babesia microti reservoir play Microtus voles. The prevalence of infection in Microtus arvalis studied in Mazurian Lakeland is 9-33%; in Microtus agrestis in Katowice agglomeration reach almost 50%, Microtus oeconomus in Bialowieza 7.7-50%. The lesser role as zoonotic reservoir play Clethrionomys voles, Apodemus mice and shrews; the prevalence of infections in these mammals don't exceed 2 %. The vectors for B. microti piroplasms in middle-European conditions are Ixodes ricinus, I. trianguliceps and Dermacentor reticulatus. There were recorded the infections of Ixodes ricinus ticks with B. microti in Szczecin and Tri-City, the rate was 6.2-13.3%. The variation in B. microti prevalence in rodents and ticks is very changeable and determined by season, the interaction with other hemoparasites, host's age and local conditions. PMID- 15787200 TI - Apicomplexan parasites: environmental contamination and transmission. AB - The Apicomplexa are a diverse group of intracellular parasitic protists. The majority of species from the classes Coccidea, Haemosporea and Piroplasmea are responsible for widespread diseases of humans and domestic animals. Oocysts of these parasites can persist for long periods of time in the environment (i.e. in water, soil, on vegetation and other food resources), maintaining their infectivity even under harsh environmental conditions and therefore are important for dispersal and transmission to hosts. This review will address the biology, transmission patterns and survival in the environment of Cryptosporidium, Cyclospora and Toxoplasma species, the most common causes of human diseases. PMID- 15787201 TI - The modulation of transferrin receptors level on mouse macrophages and fibroblasts by Toxoplasma gondii. AB - Macrophage-mediated early nonspecific immunological response is an important part of the immunity against intracellular parasite Toxoplasma gondii. The immunological functions of macrophages are closely connected with iron metabolism and acquiring of iron mainly from transferrin by the receptor-mediated endocytosis. The level of specific transferrin receptors can be modulated by different soluble exogenous and endogenous factors and also by microbial pathogens. The goal of our study was to determine the influence of T. gondii infection and toxoplasma lysate antigen (TLA) on the expression level of transferrin receptors (TfRs) on mouse macrophages and fibroblasts which can serve as host cells for the parasite replication. The level of TfRs was measured using CELISA assay. Strong down-regulation of the receptors level, started about 18 hours after infection of macrophages with a high number of freshly harvested tachyzoites T. gondii. Stimulation of the mouse cells with TLA antigen did not cause any changes in TfRs expression. In our studies we did not observe any differences in the TfRs level on mouse fibroblasts even after incubation with high concentrations of TLA antigen or inoculation with a high number of tachyzoites. PMID- 15787202 TI - [Neither with you, nor with you. Preferably with you]. PMID- 15787203 TI - [Gender differences in suicidal behavior]. AB - Gender-specific differences in suicidal behaviour have been analysed in a number of recent studies. According to these, several socioeconomic, demographic, psychiatric, familial, help-seeking differences can be identified in protective and risk factors between males and females. Gender is one of the most replicated predictors for suicide. In the framework of the WHO/EURO Multicentre Study on Suicidal Behaviour, more than fifty thousand suicide attempts have been registered so far. Until now data on more than 1200 monitored suicidal events have been collected in Pecs centre. In most countries male suicid rates are higher. In contrast to suicides, rates of suicide attempts are usually higher in females. Concerning the differences in methods, it is a recognised fact that males use violent methods of both suicide and attempted suicide more often than females. The summarised clinical impression suggests that compliance of male patients is poorer than that of females. According to our data, a typical male attempter is characterised as follows: unemployed, never married, lives alone. He tends to use violent methods; if he takes drugs, it is mostly meprobamate or carbamazepine. A lot of male attempters have alcohol problems or dependence. As for the females, we found high odds ratios in the following cases: divorced or widowed, economically inactive, depressive state in the anamnesis. Female attempters are mainly repeaters using the method of self-poisoning, mostly with benzodiazepines. As suicide is a multicausal phenomenon, its therapy and prevention should also be complex and gender differences should be taken into account in building up our helping strategies. PMID- 15787204 TI - [The connection between mentalization deficit and pragmatic language skills and neurocognition in schizophrenia]. AB - OBJECTIVE: There is an increasing interest in the background of mentalization deficit in schizophrenia. On the one hand, according to developmental psychological studies, mentalization development is connected with the development of pragmatic language skills. On the other hand, studies suggest that mentalization is dependent on the maturation of neurocognitive skills such as executive functions. Our study investigated the role of these domains in the mentalization deficit of schizophrenia. METHOD: 28 patients with schizophrenia and 20 control patients with depression took part in the first part of the study. Participants were presented first-order and second-order mentalization tasks, metaphor and irony tasks for the assessment of mentalizing skills. The pragmatic language skills were examined by "question and answer" vignettes. 20 patients with schizophrenia took part in the second part of the study. The test battery was completed with a picture recognition task and neurocognitive tests were made by all patients. RESULTS: Patients with schizophrenia performed significantly worse in the irony and pragmatic tasks, but there was no correlation between their performances in the irony and in the pragmatic tasks. Selective attention and verbal working memory showed correlation with the cumulative verbal mentalization index. Selective attention correlated with the recognition of complex mental states from pictures as well, but the successful decoding of complex mental states was dependent on the recognition of basic expressions. Attention correlated with mental state recognition as well. CONCLUSIONS: Our results suggest that the mentalization deficits of people with schizophrenia cannot be explained by pragmatic language deficits alone, and the manifest impairment is the consequence of disturbances in multiple cognitive processes. Basic neurocognitive factors such as attention, selective attention and verbal working memory can influence the mentalization skills. PMID- 15787205 TI - [Change of dopamine transporter activity (DAT) during the action of bupropion (in depression)]. AB - Bupropion has an antidepressant effect through blocking the dopamine transporter. By 99mTc-TRODAT-SPECT, we measured the baseline DAT activity of depressed patients. After 3 weeks' bupropion treatment we studied the change in DAT activity, which corresponds to the occupancy of bupropion. The average occupancy of bupropion on DAT was similar to the international findings at 20.84% in 9 depressed patients. PMID- 15787206 TI - Parkinson's disease and depression. AB - The prevalence of depression in Parkinson's disease is around 40%, but, unfortunately, such depression is frequently unrecognized and untreated. However, recognition and appropriate treatment of depression in patients with Parkinson's disease is essential for clinical practice. This review focuses on the epidemiology, pathophysiology and treatment of depression associated with Parkinson's disease. PMID- 15787207 TI - [Effect of atypical antipsychotics on metabolism]. AB - The metabolic effect of atypical antipsychotic drugs may manifest itself in weight gain, disturbances in glucose homeostasis and dyslipidemia. One confounding factor is that psychotic disorders themselves make the patients prone to specific metabolic changes. Nevertheless clinical studies have confirmed that atypical anti-psychotic drugs have a different metabolic effect. In the present prospective case-series, four male antipsychotic drug-naive psychiatric patients without any familial history of metabolic disorder were studied. The patients received risperidone or olanzapine monotherapy for 12 weeks; weight, plasma levels of triglyceride, cholesterol, HDL, and fasting glucose were measured in every 4th week. In two patients, oral glucose tolerance test (OGTT) was also performed at days 0 and 56. The average weight gain during the 12-week period was 12%. The cholesterol and triglyceride plasma concentrations were also elevated. The fasting glucose levels did not change during the observation period. In the OGTT performed in two patients, normal fasting glucose and insulin plasma levels were observed after 8 weeks; however, the plasma insulin concentrations were highly elevated after glucose intake, which may suggest the presence of insulin resistance. Our preliminary results confirmed the previous results on the metabolic effect of atypical anti-psychotic drugs, which may lead to metabolic syndrome. The regular control of the metabolic laboratory parameters, early intervention and the modification of the atypical antipsychotic treatment may help to avoid this adverse effect of the drugs. PMID- 15787208 TI - [Schizoid psychosis during cannabis intake (case report)]. AB - Three young people developed psychosis during/ after cannabis intake. The 17-year old male after only a few marihuana cigarettes, the 22-year-old patient after two years of addiction developed schizoid psychosis; the 20-year-old patient after six years of cannabis addiction had schizoaffective psychosis. The first two patients become symptom-free on the antipsychotics and during the drug-free period. The third patient, who had cannabis during the psychotic symptoms, still has the schizoid psychosis. CONCLUSIONS: The connection between cannabis and psychosis is clear in our three patients. Marihuana is working on the dopamine system and may cause schizoid psychosis, sometimes permanent psychosis. Cannabis, this light drug might not be a "safe" agent. PMID- 15787209 TI - [Solid phase isotope exchange with spillover hydrogen in amino acids, peptides, and proteins]. AB - We summarize here information on the theoretical and experimental study of high temperature (150-200 degrees C) solid phase catalytic isotope exchange (HTSPCIE) carried out with amino acids, peptides, and proteins under the action of spillover hydrogen. Main specific features of the HTSPCIE reaction, its mechanism, and its use for studying spatial interactions in polypeptides are discussed. A virtually complete absence of racemization makes this reaction a valuable preparative method. The main regularities of the HTSPCIE reaction with the participation of spillover tritium have been revealed in the case of peptides and proteins, and the dependence of reactivity of peptide fragments on the spatial organization of their molecules has been studied. An important peculiarity of this reaction is that HTSPCIE proceeds at 150-200 degrees C with a high degree of chirality retention in amino acids and peptides. This is provided by its reaction mechanism, which consists in a synchronous one-center substitution at the saturated carbon atom characterized by the formation of pentacoordinated carbon and a three-center bond between the carbon and the incoming and outgoing hydrogen atoms. PMID- 15787210 TI - [Cationic oligopeptides modified with lipophilic fragments: use for DNA delivery to cells]. AB - Cationic oligopeptides, including the amphipathic alpha-helical peptides, are applied to the targeted delivery of DNA to eukaryotic cells due to their DNA compacting properties and the ability to destabilize the cell lipid bilayer in some cases. We synthesized the peptides differing in the number and location of residues of decanoic acid covalently attached to Lys residues in order to combine the DNA-binding and the membrane activities in a single molecule. We chose peptide structures that assisted in the formation of alpha-helices. The DNA binding ability of the peptides and the membrane activity of their complexes with DNA were shown to depend on the structure. The study of erythrocyte hemolysis by complexes with DNA of the pCMV LacZ plasmid and the peculiarities of transfection of these complexes revealed a correlation between the hemolytic activity and the expression level of the lacZ gene in the cells. PMID- 15787211 TI - [Spatial structure of myelopeptides: I. Conformational analysis of MP-1, MP-2, and MP-3]. AB - Theoretical conformational analysis was used to study the spatial structure and conformational properties of myelopeptides, bone-marrow peptide mediators. The low-energy conformations of three hexapeptides MP-1 (Phe-Leu-Gly-Phe-Pro-Thr), MP 2 (Leu-Val-Val-Tyr-Pro-Trp), and MP-3 (Leu-Val-Cys-Tyr-Pro-Gln) were found, the values of dihedral angles of the backbone and side chains of the amino acid residues constituting these peptides were determined, and the energies of intra- and interresidual interactions were estimated. PMID- 15787212 TI - [Actinoporins from the Sea of Japan anemone Oulactis orientalis: isolation and partial characterization]. AB - Two cytolytic toxins (cytolysins Or-A and Or-G) were isolated from the Sea of Japan anemone Oulactis orientalis and characterized. Their purification scheme involved a hydrophobic chromatography on Polychrom 1, a gel filtration on Akrilex P-4, a cation-exchange chromatography on CM-32 cellulose, and a reversed-phase HPLC on a Nucleosil C18 column. The molecular masses of Or-A and Or-G were determined by SDS-PAGE in 14% PAG to be ca. 18 kDa. The absence of Cys residues and a high content of basic amino acid residues are characteristic of their amino acid compositions. The hemolytic activities of Or-A and Or-G were found to be 295.86 and 322.58 HU/mg, respectively; these are by three orders of magnitude lower than those of sphingomyelin-inhibitable cytolysins from the tropic sea anemones. The amino acid sequences of the N-terminal fragments of Or-A and Or-G were determined to be ATFRVLAK and GAIIAGAA, respectively. Action of the cytolysins on the erythrocyte membrane is inhibited by exogenous sphingomyelin. They form ion channels in bilayer lipid membranes with the conductivity of 16, 32, and 40 pSm in 0.1 M NaCl and 168, 240, and 320 pSm in 1 M NaCl at pH 7.2. Therefore, they were attributed to the group of actinoporins. PMID- 15787213 TI - [Spectral diversity among the members of the family of Green Fluorescent Protein in hydroid jellyfish (Cnidaria, Hydrozoa)]. AB - The cDNAs encoding the genes of new proteins homologous to the well-known Green Fluorescent Protein (GFP) from the hydroid jellyfish Aequorea victoria were cloned. Two green fluorescent proteins from one un-identified anthojellyfish, a yellow fluorescent protein from Phialidium sp., and a nonfluorescent chromoprotein from another unidentified anthojellyfish were characterized. Thus, a broad diversity of GFP-like proteins among the organisms of the class Hydrozoa in both spectral properties and primary structure was shown. PMID- 15787214 TI - [Substrate properties of dinucleoside 5',5"-oligophosphates in the reactions catalyzed by HIV reverse transcriptase, E. coli DNA polymerase I, and E. coli RNA polymerase]. AB - Dinucleoside 5',5"-oligophosphates dNp4dN', dTp3rA, dTp4rA, and dTp5rA were synthesized and evaluated as substrates for HIV reverse transcriptase, the Klenow fragment of E. coli DNA polymerase I, and E. coli RNA polymerase. A model of the HIV reverse transcriptase interaction with the dinucleoside oligophosphates was proposed and experimentally confirmed. The kinetic constants of some of the synthesized compounds for the HIV reverse transcriptase were determined. PMID- 15787215 TI - [Synthesis and antiherpetic activity of (Z)- and (E)-isomers of 9-(3 phosphonomethoxyprop-1-en-yl)adenine]. AB - 9-(3-Phosphonomethoxyprop-1-en-yl)adenine (Z)- and (E)-isomers were synthesized. The stereoselectivity of double bond formation was studied by variation of sulfonyl groups. The resulting phosphonates exhibited a moderate antiherpetic activity in a culture of Vero cells infected with herpes simplex type 1 virus. The Z-isomer was shown to be more effective inhibitor of virus reproduction in the case of both wild and acyclovir-resistant strain. PMID- 15787216 TI - [A PCR-based semiquantitative assay of DNA impurities in recombinant protein preparations]. AB - A semiquantitative assay of DNA impurities in preparations of human recombinant insulin is described. The assay is based on the detection of a fragment of the ampicillin-resistant gene within the producer strain DNA by PCR. The analysis of PCR products of the studied preparations and PCR products containing known amounts of E. coli total DNA enabled a quantitative determination of the producer strain DNA content in the preparations under study. The sensitivity of the method is 7 pg of E. coli DNA per 10 microg of human recombinant insulin. The high sensitivity of the method allows us to recommend it for the quantitative determination of DNA content in recombinant preparations that do not inhibit PCR. PMID- 15787217 TI - [Regulatory potential of S/MAR elements in transient expression]. AB - S/MARs (scaffold/matrix attachment regions) are the DNA regions that are involved in the interaction with the nuclear matrix and are identified by in vitro methods. According to the available information, S/MARs possess an insulating activity, i.e., the ability to block the interaction between the enhancer and promoter in vivo, and are, probably, intact insulators or their fragments. Nevertheless, there is still no direct proof for this correspondence. To obtain additional information on the insulator activity of S/MARs, we selected five DNA fragments of different lengths and affinities for the nuclear matrix from the previously constructed library of S/MARs and tested their ability to serve as insulators. Two of five elements exhibited an insulator (enhancer-blocking) activity upon the transient transfection of CHO cells. None of the S/MARs displayed either promoter or enhancer/silencer activities in these cells. PMID- 15787218 TI - [Multiple template switches on LINE-directed reverse transcription: the most probable formation mechanism for the double and triple chimeric retroelements in mammals]. AB - It was shown that the shuffling mechanism for transcribed genome components, which involves a template switch on the RNA reverse transcription using the L1 retroelement enzymatic machinery, is common in mammals. The occurrence frequency of the resulting chimeric retroelements in the genomes of rodents is twice as high as in the DNA of primates. Moreover, we proved that not only single but also double switches may occur in vivo, which result in the fusion of copies of three different transcripts. Many of the identified chimeras are transcribed in mammals. PMID- 15787219 TI - [Modification of hyaluronic acid with aromatic amino acids]. AB - Hyaluronic acid was modified with aromatic amino acids (5-aminosalicylic, 4 aminosalicylic, anthranilic, and p-aminobenzoic) in the presence of 1-ethyl-3-[3 (dimethylamino)propyl]carbodiimide. The modified glycans contained 9-43% of arylamide groups and 10-33% of isoureidocarbonyl groups depending on the nature of the amino acid. Reduction with sodium borohydride allowed the conversion of isoureidocarbonyl groups into hydroxymethyl groups. PMID- 15787221 TI - [Formylation of porphyrin platinum complexes]. AB - The formylation reaction of platinum complexes of beta-unsubstituted porphyrins was studied. The interaction of deuteroporphyrin IX derivatives with the Vilsmeyer reagent led to the selective formylation of their macrocycles in the beta position. The resulting formyl derivatives of the porphyrins are of interest for fluorescent immunoassay. PMID- 15787220 TI - [The synthesis of imidodiphosphoric acid derivatives as potential substrates in the pyrophosphorolysis reaction]. AB - The preparation conditions for dichlorophosphinylphosphorimidic trichloride were optimized. It was used in the synthesis of esters of imidodiphosphoric acid. The interaction of the trichloride with amines resulted in the corresponding amidodiphosphates rather than in the expected amides of imidodiphosphoric acid. PMID- 15787222 TI - [An express morphine assay in aqueous samples by immunochromatography using monoclonal antibodies labeled with colloidal gold]. AB - A simple and accessible express method was developed for the detection of morphine in aqueous samples using competitive immunochromatography. A complex of colloidal gold with monoclonal antibodies to morphine was used as the detection agent. The detection limit for morphine in aqueous samples was 10 ng/ml, and the analysis time was 5 min. PMID- 15787223 TI - Patients offered pick of any ITC or foundation by 2006. PMID- 15787224 TI - PCTs must work alongside councils. PMID- 15787225 TI - Surgeons fear FoI-empowered media will end up firing blanks. PMID- 15787226 TI - On partners in commissioning. PMID- 15787227 TI - 'Monitor would do things differently today' [interview by Helen Mooney]. PMID- 15787228 TI - Data briefing. Length of stay. Discharge variations for emergencies. PMID- 15787229 TI - Primary care. On the mend? AB - Alternative provider medical services contracts are encouraging primary care trusts to innovate in the way they invite tenders--but private companies have had mixed success in winning business. However, now the situation seems to be changing, with about half of all out-of-hours contracts run under APMS rules and a small number of prison and general practice 'wins'. PMID- 15787230 TI - On distorting diagrams. PMID- 15787231 TI - Clinical management. Sexual frustration. AB - West Hull PCT has set up a joint sexual health an GUM clinic, echoing the inter disciplinary principles outlined in the public health white paper. Other PCTs have found blurred skill boundaries and training problems to be huge barriers to the policy. Some trusts are "not even close" to meeting the DoH's 48-hour GUM access target by the 2008 deadline. PMID- 15787232 TI - Education. Out of the age of Aquarius. PMID- 15787233 TI - Sickness absence. Help staff with illness. PMID- 15787234 TI - Immigration. What migrants do for the NHS. PMID- 15787235 TI - 6-Substituted indanoyl isoleucine conjugate induces tobacco plant responses in secondary metabolites. AB - To characterize the role of the phytotoxin mimic 6-substituted indanoyl isoleucine conjugate 1 in plant secondary metabolism, tobacco (Nicotiana tabacum L. K326) was treated with compound 1. The volatile compounds of tobacco leaves were analyzed by GC-MS. In contrast to the control, three compounds, farnesene (2), santalol (3) and tetradecanal (4), were induced by treatment with 1 mM of compound 1. Concurrently other volatile compounds were also regulated. PMID- 15787236 TI - Secondary metabolite content in rhizomes, callus cultures and in vitro regenerated plantlets of Solidago chilensis. AB - An in vitro culture system leading to the formation of callus and plant regeneration, starting from nodal sections and shoot tips, was developed for Solidago chilensis (Asteraceae). The content of the gastroprotective diterpene solidagenone as well as the phenolics chlorogenic acid (CA) and rutin was determined either in rhizomes from wild growing plants and in callus and in in vitro regenerated plantlets by analytical HPLC. Additionally, total phenolic and flavonoid content was assessed in plant samples, callus and cell suspensions. In terms of dry starting material, the percentual solidagenone content in nine S. chilensis samples ranged from 0.5-3.5% for rhizomes from wild growing plants, 0.1 0.3% for callus and 0.3% for an in vitro regenerated plantlet, respectively. The highest solidagenone contents were found in the wild plant during the late summer in the months of March and April (3.5-2.2%) while highest values for chlorogenic acid (0.5%) and rutin (0.4%) were detected in May, before senescence. The callus tissue and cell suspensions contained some 1.8-2.0 and 1.2% of total phenolics, respectively. CA was the main phenolic in the cell suspension while only traces were found in the callus. Rutin was not detected in the callus nor cell culture. PMID- 15787237 TI - Metabolites from Microsphaeropsis olivacea, an endophytic fungus of Pilgerodendron uviferum. AB - Seven compounds belonging to different structural skeletons were isolated from Microsphaeropsis olivacea grown in liquid and solid media. The enalin derivative 7-hydroxy-2,4-dimethyl-3(2H)-benzofuranone is reported for the first time, while additional spectroscopic information is provided for the acetates of botrallin and ulocladol. The activity of the isolated compounds was assessed towards the enzyme acetylcholinesterase (AChE) and their cytotoxicity against human lung fibroblasts. Graphislactone A and botrallin presented a moderate activity towards AChE, with IC50 of 8.1 and 6.1 microg/ml (27 and 19 microM, respectively). Under the same experimental conditions, the IC50 of the standard inhibitor galanthamine was 3 microg/ml. The cytotoxicity of both compounds was > 1000 and 330 microM, respectively. None of the compounds was promising as antibacterial or antifungic against phytopathogenic fungi and bacteria. Botrallin and graphislactone A were detected in the liquid potato-dextrose and yeast extract/malt extract/dextrose as well as on a solid substrate (rice). Butyrolactone I was obtained from the fungus growing on solid medium. PMID- 15787238 TI - Bioactive constituents from Dracocephalum subcapitatum (O. Kuntze) Lipsky. AB - From an EtOAc extract of Dracocephalum subcapitatum, five flavonoids, calycopterin, xanthomicrol, isokaempferide, luteolin and apigenin, together with five terpenoids, oleanolic acid, ursolic acid, geranial, neral and limonene-10 al, were isolated. Among them, citral and limonene-10-al were the most effective components against epimastigotes of Trypanosoma cruzi, the parasitic agent of Chagas disease. PMID- 15787240 TI - Antibacterial activity and chemical composition of the essential oil of Grammosciadium platycarpum Boiss. from Iran. AB - The chemical composition of the essential oils obtained from two samples (GP1 and GP2) of Grammosciadium platycarpum Boiss. was analyzed by GC and GC-MS. The analysis of the oils resulted in the identification of twenty-two constituents. Linalool (79.0%-GP1, 81.8%-GP2) and limonene (10.0%, 5.8%) were found to be the major components, respectively. The in vitro antibacterial activities of these oils and their main compounds against seven Gram-positive and Gram-negative bacteria were investigated. The results exhibited that the total oils and their major components possess strong to moderate activities against all the tested bacteria except for Pseudomonas aeruginosa. PMID- 15787239 TI - Gas chromatography-mass spectrometry study of the essential oils of Schinus longifolia (Lindl.) speg., Schinus fasciculata (Griseb.) I. M. Johnst., and Schinus areira L. AB - The essential oil composition from the aerial parts of three Anacardiaceae growing in Bahia Blanca, Argentina was studied by gas chromatography and gas chromatography-mass spectrometry. The essential oils of S. longifolia and S. fasciculata have been studied for the first time. The major constituents were alpha-pinene (46.5%), beta-pinene (15.1%) and alpha-phellandrene (10.1%) for S. longifolia and limonene (10.9%), beta-phellandrene (6.16%) and alpha-phellandrene (5.6%) for S. fasciculata. The major components of the essential oil of S. areira were limonene (28.6%), alpha-phellandrene (10.1%), sabinene (9.2%) and camphene (9.2%) differing from the literature data. The essential oils from S. areira and S. longifolia exhibited a high biotoxicity in a brine shrimp assay with Artemia persimilis. PMID- 15787241 TI - The antimicrobial activity of extracts of the lichen Hypogymnia tubulosa and its 3-hydroxyphysodic acid constituent. AB - The antimicrobial activity and the MIC values of the diethyl ether, acetone, chloroform, petroleum ether, and ethanol extracts of the lichen Hypogymnia tubulosa and its 3-hydroxyphysodic acid constituent have been investigated against some microorganisms. At least one of the extracts or 3-hydroxyphysodic acid showed antimicrobial activity against Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Listeria monocytogenes, Proteus vulgaris, Salmonella typhimurium, Staphylococcus aureus, Streptococcus faecalis, and Candida albicans. No antifungal activity of the extracts has been observed against ten filamentous fungi. PMID- 15787242 TI - Antimicrobial substances from rhizomes of the giant knotweed Polygonum sachalinense against the fish pathogen Photobacterium damselae subsp. piscicida. AB - The antimicrobial compounds against the fish pathogen Photobacterium damselae subsp. piscicida were isolated from Polygonum sachalinense rhizomes. The structures of the antimicrobial compounds 1 and 2 were determined by 1H and 13C NMR, 2D-NMR (COSY, HSQC, HMBC and ROESY) and FAB-MS to be phenylpropanoid glycosides, vanicoside A and B, respectively. Both compounds have feruloyl and p coumaroyl groups bonded to a sucrose moiety in their structures. Vanicoside A also has an acetyl group in the sucrose moiety. The MIC values for vanicoside A and B against Ph. damselae subsp. piscicida DPp-1 were 32 and 64 microg/ml, respectively. The antimicrobial activities of these vanicosides were modest, in contrast to higher activities (MICs at < 4 microg/ml) of antibiotics, florphenicol, ampicillin and amoxicillin, which have been generally used for treating pasteurellosis. The activities of the vanicosides, however, were higher than those (MICs at 256 microg/ml) of ferulic acid and p-coumaric acid. It was suggested that the structure of phenylpropanoids esterified with sucrose was essential for higher antimicrobial activity of vanicosides and also acetylation of sucrose might affect the activity against the bacterium. PMID- 15787243 TI - A new flavonol glycoside derivative from leaves of Moldenhawera nutans. AB - The ethyl acetate extract of leaves of Moldenhawera nutans Queiroz & Alkin (Leguminosae) furnished, besides methyl gallate and gallic acid, the flavonols named laricetrin, laricetrin 3-glucoside and laricetrin 3-galactoside as well as the new one named laricetrin 5-galloyl-3-beta-D-xylopyranoside. It also was isolated from the hexane extract: beta-sitosterol, lupenone, beta-amyrinone, alpha-amyrinone, lupeol, beta-amyrin, alpha-amyrin and alpha-tocopherol. The antioxidant activities of flavonoids were measured through DPPH radical scavenging and inhibition of auto-oxidation of beta-carotene methods. The structures of the compounds were determined by analyses of spectral data. This is the first report dealing with phytochemical studies of leaves of M. nutans. In addition this current work describes the unequivocal attribution of 1H NMR and 13C NMR data of laricetrin. PMID- 15787244 TI - Activity in vitro and in vivo against plant pathogenic fungi of grifolin isolated from the basidiomycete Albatrellus dispansus. AB - In the course of screening for novel naturally occurring fungicides from mushrooms in Yunnan province, China, the ethanol extract of the fruiting bodies of Albatrellus dispansus was found to show antifungal activity against plant pathogenic fungi. The active compound was isolated from the fruiting bodies of A. dispansus by bioassay-guided fractionation of the extract and identified as grifolin by IR, 1H and 13C NMR and mass spectral analysis. Its antifungal activities were evaluated in vitro against 9 plant pathogenic fungi and in vivo against the plant disease of Erysiphe graminis. In vitro, Sclerotinina sclerotiorum and Fusarium graminearum were the most sensitive fungi to grifolin, and their mycelial growth inhibition were 86.4 and 80.9% at 304.9 microM, respectively. Spore germination of F. graminearum, Gloeosporium fructigenum and Pyricularia oryzae was almost completely inhibited by 38.1microM grifolin. In vivo, the curative effect of grifolin against E. graminis was 65.5% at 304.9 microM after 8 days. PMID- 15787245 TI - A new bioactive steroidal saponin from Agave shrevei. AB - A new steroidal saponin was isolated from the leaves of Agave shrevei Gentry. Its structure was established as 26-(beta-D-glucopyranosyloxy)-22-methoxy-3-(O-beta-D glucopyranosyl-(1-->2)O-[O-beta-D-glucopyranosyl-(1-->4)-O-[O-beta-D glucopyranosyl-(1-->6)]-O-beta-D-glucopyranosyl(1-->4)-beta-D galactopyranosyl]oxy)-(3beta,5alpha,25R)-furostane. The structural identification was performed using detailed analyses of 1H and 13C NMR spectra including 2D NMR spectroscopic techniques (COSY, HETCOR, and COLOC) and chemical conversions. The steroidal saponin showed absence of haemolytic effects in the in vitro assay, but demonstrated a significant inhibition of the capillary permeability activity. PMID- 15787246 TI - Topical anti-inflammatory activity of flavonoids and a new xanthone from Santolina insularis. AB - Bioactivity-guided fractionation of the methanol extract from the leaves of Santolina insularis led to the isolation of one new xanthone, (E)-3-(6-[(E)-3 hydroxy-3-oxo-1-propenyl]-9-oxo-9H-xanthen-2-yl)-2-propenoic acid, together with six known flavonoids: hispidulin, nepetin, cirsimaritin, rhamnocitrin, luteolin and luteolin 7-O-beta-D-glucopyranoside. The structures were elucidated by means of 1D-, 2D-NMR spectroscopy and mass spectrometry. The topical anti-inflammatory activity of all isolated compounds and extracts was investigated employing the croton oil-induced dermatitis in mouse ear. The most active compound, luteolin, showed an ID50 of 0.3 micromol/cm(2) and prevented ear oedema more effectively than an equimolar dose of indomethacin within 24 h. PMID- 15787247 TI - Spasmolytic flavonoids from Syzygium samarangense (Blume) Merr. & L.M. Perry. AB - The hexane extract of Syzygium samarangense (Ss.Hex) dose-dependently (10-1000 microg/ ml) relaxed the spontaneously contracting isolated rabbit jejunum. Four rare C-methylated flavonoids with a chalcone and a flavanone skeleton were isolated from Ss.Hex and were subsequently tested for spasmolytic activity. All flavonoids, identified as 2'-hydroxy-4',6'-dimethoxy-3'-methylchalcone (1), 2',4' dihydroxy-6'-methoxy-3',5'-dimethylchalcone (2), 2',4'-dihydroxy-6'-methoxy-3' methylchalcone (3), and 7-hydroxy-5-methoxy-6,8-dimethyl-flavanone (4), showed dose-dependent spasmolytic activity in the rabbit jejunum with IC50 values of 148.3 +/- 69.4, 77.2 +/- 43.5, 142.4 +/- 58.6 and 178.5 +/- 37.5 microg/ml (mean +/- SEM), respectively. The dihydrochalcone derivative of compound 1, 2'-hydroxy 4',6'-dimethoxy-3'-methyldihydrochalcone (5), when tested for spasmolytic activity, did not significantly relax the smooth muscle relative to the other compounds. Verapamil, a standard spasmolytic, has an IC50 value of 0.16 +/- 0.04 microg/ml. This is the first report of the relaxant activity of chalcones, specifically of compounds 1-3. PMID- 15787248 TI - Effects of kaurane diterpene derivatives on germination and growth of Lactuca sativa seedlings. AB - Kaurenoic and grandiflorenic acid, isolated from Wedelia paludosa (Asteraceae), some derivatives from these acids (alcohols, esters, amides, lactones, oximes) and other naturally occurring kaurane diterpenes were tested for their action on the growth of radical and shoot of Lactuca sativa. Gibberellic acid, GA3, a commercially available phytohormone, belonging to the same class of diterpenes, was also tested. Some of the tested substances showed a remarkable activity either in the inhibition or in stimulation of L. sativa growth. The activity, in some cases, was even higher than that of GA3. PMID- 15787249 TI - Germination inhibitor from the Japanese cedar Cryptomeria japonica. AB - (1S,6R)-2,7(14),10-Bisabolatrien-1-ol-4-one was identified as a germination inhibitor from the methanol extract of Japanese cedar wood, Cryptomeria japonica. The occurrence of this compound in 1 g fresh wood was 2.0 mg, and showed a maximum of 60% germination inhibition at the dose of 20 mg/filter paper (157 microg/cm(2)) against both of lettuce and rice seeds for 4 d. A selective activity between Dicotyledoneae and Monocotyledoneae seeds was not observed. PMID- 15787250 TI - Morning glory systemically accumulates scopoletin and scopolin after interaction with Fusarium oxysporum. AB - An isolate of non-pathogenic Fusarium, Fusarium oxysporum 101-2 (NPF), induces resistance in the cuttings of morning glory against Fusarium wilt caused by F. oxysporum f. sp. batatas O-17 (PF). The effect of NPF on phenylpropanoid metabolism in morning glory cuttings was studied. It was found that morning glory tissues responded to treatment with NPF bud-cell suspension (108 bud-cells/ml) with the activation of phenylalanine ammonia-lyase (PAL). PAL activity was induced faster and greater in the NPF-treated cuttings compared to cuttings of a distilled water control. High performance liquid chromatography analysis of the extract from tissues of morning glory cuttings after NPF treatment showed a quicker induction of scopoletin and scopolin synthesis than that seen in the control cuttings. PF also the induced synthesis of these compounds at 10(5) bud cells/ml, but inhibited it at 10(8) bud-cells/ml. Possibly PF produced constituent(s) that elicited the inhibitory effect on induction of the resistance reaction. These compounds could potentially be useful as markers to detect early beginning interactions between Fusarium and morning glory tissues cuttings. PMID- 15787251 TI - Purification and biochemical characterization of polygalacturonases produced by Aureobasidium pullulans. AB - The extracellular polygalacturonases produced by Aureobasidium pullulans isolated from waters of the Danube river were partially purified and characterized. The pH optima of polygalacturonases produced in the first phases of cultivation (48 h) and after 10 d as well as their optima of temperature, thermal stabilities, molecular masses, isoelectric points, action pattern and ability to cleave polymeric and oligomeric substrates were compared. Polygalacturonases with a random action pattern (random cleavage of pectate forming a mixture of galactosiduronides with a lower degree of polymerization) [EC 3.2.1.15] were produced only in the first phases of growth, while exopolygalacturonases [EC 3.2.1.67] with a terminal action pattern (cleavage of pectate from the nonreducing end forming D-galactopyranuronic acid as a product) were found during the whole growth. The main enzyme form with a random action pattern was glycosylated and its active site had the arrangement described previously for the active site of polygalacturonase of phytopathogenic fungi. PMID- 15787253 TI - Transformations of steroids by Beauveria bassiana. AB - The course of transformations of testosterone and its derivatives, including compounds with an additional C1,C2 double bond and/or a 17alpha-methyl group, a 17beta-acetyl group or without a 19-methyl group, by a Beauveria bassiana culture was investigated. The fungi promoted hydroxylation of these compounds at position 11alpha, oxidation of the 17beta-hydroxyl group, reduction of the C1,C2 or C4,C5 double bonds and degradation of the progesterone side-chain, leading to testosterone. The structure of 4-ene-3-oxo-steroids had no influence on regio- and stereochemistry of hydroxylation. In a similar manner, dehydroepiandrosterone was hydroxylated by Beauveria bassiana at position 11alpha, however, a small amount of 7alpha-hydroxylation product was also formed. PMID- 15787252 TI - Degradation of aromatic compounds in plants grown under aseptic conditions. AB - The aim of the work is to investigate the ability of higher plants to absorb and detoxify environmental pollutants - aromatic compounds via aromatic ring cleavage. Transformation of 14C specifically labelled benzene derivatives, [1-6 14C]-nitrobenzene, [1-6-(14)C]-aniline, [1-(14)C]- and [7-(14)C]-benzoic acid, in axenic seedlings of maize (Zea mays L.), kidney bean (Phaseolus vulgaris L.), pea (Pisum sativum L.) and pumpkin (Cucurbita pepo L.) were studied. After penetration in plants, the above xenobiotics are transformed by oxidative or reductive reactions, conjugation with cell endogenous compounds, and binding to biopolymers. The initial stage of oxidative degradation consists in hydroxylation reactions. The aromatic ring can then be cleaved and degraded into organic acids of the Krebs cycle. Ring cleavage is accompanied by 14CO2 evolution. Aromatic ring cleavage in plants has thus been demonstrated for different xenobiotics carrying different substitutions on their benzene ring. Conjugation with low molecular peptides is the main pathway of aromatic xenobiotics detoxification. Peptide conjugates are formed both by the initial xenobiotics (except nitrobenzene) and by intermediate transformation products. The chemical nature of the radioactive fragment and the amino acid composition of peptides participating in conjugation were identified. PMID- 15787254 TI - Purification and partial characterization of an extracellular melanoprotein from the fungus Venturia inaequalis. AB - The fungus Venturia inaequalis clone No. 36 isolated from Malus domestica cv. Gloster excretes a melanoprotein of 36 kDa in relatively high amounts during growth in liquid culture. The protein was isolated from the culture medium and purified to homogeneity. It was shown to contain melanin. After raising an antiserum against the isolated protein, the protein could be shown to be located in the apoplast fluid of the V. inaequalis infected Malus domestica cv. Elstar. Partial sequencing of the protein revealed no significant sequence homologies to so far sequenced proteins. The melanoprotein binds ferrous and ferric iron. Moreover, it could be shown that the binding of ferric iron (but not of ferrous iron) leads to a change in the absorbance of the protein suggesting a modification of the protein by ferric, but not by ferrous, iron. In addition to iron, the protein also binds copper, but does not bind manganese or nickel. A possible function of this protein in the recruiting and transport of iron and copper and/or in the protection of the fungus by metal-ion mediated oxidative stress is discussed. PMID- 15787255 TI - Pigments and citrinin biosynthesis by fungi belonging to genus Monascus. AB - Citrinin is a mycotoxin, which is produced by fungi belonging to the genus Monascus, known in biotechnology as producers of azaphilone pigments. The relation between biosynthesis of these secondary metabolites was investigated in different species of the genus Monascus in batch-culture at the following cultivation conditions: T = 28 degrees C, agitation 220 rpm, and a medium, which induce citrinin production, containing ethanol as a carbon source. The screening was carried out with 16 fungal strains and the biosynthesis of citrinin and pigments was monitored quantitatively at the standard conditions mentioned above. Some kinetic parameters of the process have been determined. The values of the growth yield coefficient Y(X/C) were between 0.32 and 0.57. The amount of the extracellular red and orange pigments at the end of cultivation varied for the different strains between 0.09 and 1.33 OU/ mg dry weight, and 0.15 and 0.96 OU/mg dry weight, respectively. The amount of the total pigments measured was between 0.16 and 3.6 OU/mg dry weight, and between 0.21 and 3.39 OU/mg dry weight. The determined ratio 500 nm/400 nm, characterizing the pigment production, ranged between 0.60 and 1.06. Twelve of the investigated strains produced citrinin and pigments, two of them produced only pigments. Two strains were not able to produce neither pigments nor citrinin. Thus, the biosynthesis of citrinin appeared to be strain-specific and does not correlate with the pigments' biosynthesis by the fungal strains belonging to the genus Monascus. PMID- 15787256 TI - A new steroidal saponin from AGAVE brittoniana and its biphasic effect on the Na+ ATPase activity. AB - A new steroidal saponin, 3-{(O-6-deoxy-a-L-mannopyranosyl-(1 --> 4)-O-beta-D glucopyranosyl-(1 --> 3)-O-[O-beta-D-glucopyranosyl-(1 --> 3)-beta-D glucopyranosyl-(1 --> 2)]-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D galactopyranosyl)oxy}-6-hydroxy-(3beta,5alpha,6alpha,25R)-spirostan-12-one, was isolated from Agave brittoniana Trel. The structure was determined by extensive NMR spectroscopy studies and chemical conversions. Its effects on the Na+-ATPase and (Na+ + K+)-ATPase activities of the proximal tubule from pig kidney were evaluated. It was observed that this steroidal saponin exerts a biphasic effect on the Na+-ATPase activity. It is concluded that the effect of the aqueous extract as a diuretic is due, at least in part, to the action of saponin on the ouabain-insensitive Na+-ATPase. PMID- 15787257 TI - Changes in phenylalanine ammonia-lyase activity and gene expression during storage of asparagus spears. AB - A cDNA clone coding phenylalanine ammonia-lyase (PAL) was isolated from a cDNA library prepared from asparagus spears (Asparagus officinalis L. cv. Welcome) using the reverse transcription-polymerase chain reaction (RT-PCR). The partial cDNA clone encoded an mRNA of 527 bp and the derived amino acid sequence was found highly homologous to PAL from rice, maize and barley. Northern blot analysis showed an increase of pAS-PAL mRNA until 24 h at 20 degrees C, which coincided well with PAL activity and fiber development, suggesting that the increase is a response to the wounding associated with harvest. PMID- 15787258 TI - Antioxidant activity and angiotensin I-converting enzyme inhibition by enzymatic hydrolysates from bee bread. AB - Enzymatic hydrolysates were prepared from bee bread using three proteases. The antioxidant properties of these hydrolysates were measured using four different methods. These had remarkable antioxidant activity similar or superior to that of 1 mM alpha-tocopherol. They also had high scavenging activities against active oxygen species as the superoxide anion radical and hydroxyl radicals. Moreover, they showed angiotensin I-converting enzyme inhibitory activities and the activities were similar to those from various fermented foods such as fish sauce, sake, vinegar, cheese, miso, and natto. The present studies reveal that enzymatic hydrolysates from bee bread are of benefit not only for the materials of health food diets, but also for in patients undergoing various diseases such as cancer, cardiovascular diseases, diabetes, and hypertension. PMID- 15787259 TI - In vitro study of flavonoids, fatty acids, and steroids on proliferation of rat hepatic stellate cells. AB - There is a wealth of evidence that hepatic stellate cells (HSCs) orchestrate most of the important events in liver fibrogenesis. After liver injury, HSCs become activated to a profibrogenic myofibroblastic phenotype and can regulate net deposition of collagens and other matrix proteins in the liver. The proliferation of HSCs is mainly stimulated by the platelet-derived growth factor (PDGF). In this study, some compounds from natural resources have been tested for their activity to inhibit PDGF-driven proliferative activity of rat HSCs. Apigenin, quercetin, genistein, daidzin, and biochanin A exhibited > 75% inhibitory activity against HSC-T6. It was found that, gamma-linolenic (gamma-Ln), eicosapentanoic (EPA) and a- linolenic (alpha-Ln) acids showed a high inhibitory effect on proliferation of rat HSCs at 50 nmol/1. Cholest-4-ene-3,6-dione and stigmastone-4-en-3,6-dione are the most active steroids with inhibitory activities > 80% and this is most likely due to the presence of the 4-en-3,6 dione moiety in both compounds. These results revealed that the compounds which effectively blocked HSC proliferation may be beneficial in liver fibrosis. Structure-activity relationships (SAR) may provide a basis for rational structure modification. PMID- 15787260 TI - Comparison of different procedures for the lipid extraction from HL-60 cells: a MALDI-TOF mass spectrometric study. AB - A human leukaemia cell line--HL-60--can be differentiated into neutrophils or macrophages and both differentiation processes are accompanied by changes of the lipid composition. Various methods were described for the extraction of lipids from cellular systems, but only two of them were applied to the HL-60 cell line so far. In this study we compared five selected extraction methods for the lipid extraction from HL-60 cells with regard to their qualitative analysis by matrix assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS): chloroform/methanol at volume ratios 2:1 and 1:2, isopropanol/ chloroform, isopropanol/hexane and butanol. In addition, the cholesterol and phospholipid concentrations in organic extracts were measured by colorimetric assays. Results can be summarized as follows: For the analysis of polar phospholipids obtained from HL-60 cells by MALDI-TOF MS, a chlorofom/methanol (1:2) or isopropanol/chloroform mixture or butanol can be applied as extraction systems On the other hand, if one would like to analyze changes in triacylglycerols, then chloroform/methanol (2:1) would be the method of choice. PMID- 15787261 TI - [Scrotal elephantiasis: about seven operated cases]. AB - The authors report seven cases of scrotal elephantiasis operated on nine years in the urology department of a filarial endemic zone. Patients wait for a long time before consulting for big scrotum reaching eventually 15 kg. The final arguments for the filarial origin haven't been found whereas arguments of presumption exist The surgical treatment has enabled to get an aesthetic satisfying result, sometimes after a second cure. PMID- 15787262 TI - [Immunophenotyping of acute leukemias: diagnostic and pronostic utility in Abidjan, Cote d'Ivoire]. AB - Flow cytometry is nowadays the first-line method for immunophenotypic identification of blast cells but is not so usual in limited-resources countries. We have investigated on the usefulness of this tool in Abidjan, Cote d'Ivoire. Bone marrow sample from 13 patients with acute leukemia identified by cytology and cytochemical analysis was immunophenotyped by using monoclonal antibodies directed to: T lymphoid cells (CD3, CD5, CD7); B lymphoid cells (CD10, CD19, CD20, CD22, HLA-DR) and myeloid cells (CD13, CD33). Immunophenotyping allowed us to confirm the diagnosis of 6 de novo acute leukemias (2 acute myeloid leukaemias, 4 acute lymphoid leukemias) and 7 acute leukaemias resulting from chronic myeloid leukaemias. Immunophenotyping also characterizes the atypical/aberrant lineage essential for the prognosis: 2 biphenotypic acute leukemias (myeloid/lymphoid T) were identified. Our results suggest that flow cytometry may be a useful additional tool to identify the specific leukemic cell, to make a better classification as well as a prognosis evaluation of patients with acute leukemias. PMID- 15787263 TI - [Digestive bleeding and anti-inflammatory drugs at the teaching hospital of Brazzaville]. AB - Authors relate a retrospective survey of 140 cases of digestive haemorrhage after intake of anti-inflammatory drugs from January 1988 to December 2000. The digestive haemorrhage frequency after intake of anti-inflammatory drugs reaches 16%. This survey included 94 men and 46 women whose average age is 42.3 years, with extremes of 15 and 79 years. Among these patients, 75 took salicylic acetyl acid (SAA), 56 non-steroidal anti-inflammatory drug (NSAID) and 9 the SAA and NSAID association. More than half of patients relied on self-medication (91 cases ie. 65%). The haematemesis followed by melaena induced the most frequent bleeding (42.8% of cases), while the isolated haematemesis, the isolated melaena and the isolated rectal haemorrhage were found respectively in 35%, 15% and 7.2%. Haemorrhages occur early under SAA and seem less severe than those occurring under NSAID. Lesions responsible for the bleeding are: the duodenal ulcer (68.7%), the haemorrhagic gastritis (6.4%), the gastric ulcer (5%), the oesophagitis (4.2%), the gastric and duodenal ulcer association (3.5%), anorectal exsudative pains and haemorrhagic anal ulcerations. The duodenal ulcer predominance could be attributed to previous lesions. PMID- 15787264 TI - [Blackwater fever during antimalarial treatment in Abidjan (West Africa): report of 41 cases]. AB - Intravascular haemolysis, particularly blackwater fever is a rare but severe clinical syndrome, occurring after ingestion of antimalarials. A resurgence of this affection which occurred frequently during the colonization has lately been noticed. We have conducted a prospective study in order to identify the main antimalarials which are responsible for this syndrome. We reported 41 cases from 1996 to 2000, among which 80% of blackwater fever cases were associated with quinine ingestion or similar structural molecules. Their causal role is well established. For the other molecules it is difficult to underscore their role. The mortality rate is around 18%. The morbidity is high because 90% of patients have suffered from renal failure. Among them, 47% required dialysis. We are facing a public health issue thus a rational use of antimalarials is necessary. PMID- 15787265 TI - [Toward strengthening the health politics in Africa: the military health system and its contribution to health policy in Senegal]. AB - Since the following days of independences, the Senegalese army mission has mainly consisted in defending the national territory integrity and in ensuring the protection of the populations and their goods. In the public health system, thanks to the quality of its human resources the army intervenes specifically at every level of the health care structure. The mission assigned to the Senegalese army health unit is therefore multidimensional. In 2001, the operational budget of the army health services is estimated at 177 millions CFA F (265500 euros), its medical consumption at 212 millions CFA F (323 000 Euros) and its health expenditure at 385 millions CFA F (585 000 euros). The army supports the government health policies in different ways: on the one hand, availability of the ministry of health staff, on the other hand, the direct involvement in health care and the implementation of the national and international health programmes. PMID- 15787266 TI - [Prevalence of HIV infection in patients of the anti-venereal center of the National Institute of Public Hygiene, Abidjan]. AB - This cross-sectional study includes all new patients consulting for the first time in the anti-venereal center of the National Institute of Public Health (NIPH) during the period of May to July 1997. Among them, 20.1% of the consulting patients and 31.3% of those suffering from STI were positive to HIV. The STI particularly those with ulceration are a risk factor in HIV transmission. This study points up the need of an adequate management of sexual transmitted diseases as a main strategy to struggle against HIV/AIDS infection. PMID- 15787267 TI - [Epidemiological, clinical and biological profile of resistant or recurrent pulmonary tuberculosis in Abidjan]. AB - From August to December 1998, over a period of 5 months, a cross-sectional study had been carried out in Abidjan in order to analyze the epidemiological and microbiological features of BAAR+ TB patients in situation of failure or relapse after specific treatment. We investigated 79 patients enrolled in the departments of pulmonary disease of two general hospitals in Abidjan (CHU of Cocody and Treichville) and a TB outpatients' clinic. From 45 strains of Mycobacterium obtained by culture, 33 antibiograms were performed. The rate of multi-drug resistance (MDR-TB) was 79%. Among MDR--TB patients, those aged of 20-40 years were the most concerned age group (72%) with a clear male predominant rate (sex ratio: 3). Among them 49/79 (62%) had an educational level lower or equal to primary school standarts and most of them lived in dwellings with common yard (67%). In their medical history only 40% had tuberculosis and 2 cases of self medication were reported. MDR--TB prevailed among patients having at first a positive bacilloscopy. No link between HIV infection and MDR--TB was found. PMID- 15787268 TI - [Medical transportation of Congolese children by the Foundation "Terre des hommes" Netherlands (1989--1998)]. AB - We report the 10 year assessment of collaboration with the Foundation "Terre des hommes" concerning the medical transfer in the Netherlands of 41 children carrying pathologies which couldn't be treated or operated on in Brazzaville. The average age was 3 years and 6 months old (extremes: 2 months - 15 years). 33 non cyanotic cardiopathies dominated by ventricular septal defect (VSD) (n = 10) and 11 cyanotic cardiopathies among them the tetralogy of Fallot (n = 5) were admitted. The other pathologies were respectively: osseous (n = 3), vesical (n = 2), pulmonary tumoral, ophthalmic in 1 case. The surgery consisted in a complete repair in 19 cases, palliative in 9 cases. Two children proved to be inoperable. Eight other extra-cardiac pathologies had a specific surgery for each case. The average stay in the Netherlands was 1 month 13 days (extremes: 1 - 12 months). The evolution was favourable for 35 children all pathologies included. Four deaths occurred in the Netherlands and 2 in Congo. The organization of the technology transfer would be probably a better choice in the future. PMID- 15787269 TI - [Severe malaria in native adults in Abidjan (Cote d'Ivoire)]. AB - Our retrospective study carried out from 1985 to 1998 in the Unit of Infectious Diseases in Abidjan aimed at describing the epidemiological, clinical and prognosis features of severe malaria among native adults. Within 14 years, we have listed 274 cases of severe malaria for 54 098 hospitalizations (0.5%). 164 men and 110 women were recorded (sex-ratio = 1.5), aged of 33 years (16-86), among them 48% were HIV positive. 23% of the patients had already received an antimalarial treatment. The main clinical presentation was cerebral malaria (78%). The other manifestations were respiratory symptoms (13%), kidney failure (11%), anaemia (11%), macroscopic haemoglobinuria (6%), hypoglycaemia (9%), cardiovascular shock (4%). The average parasite load in blood was 27 222 plasmodium/microl (25 000 - 180200). The treatment used was quinine IV (172 patients), and arthemeter (102 patients). The outcome was favourable in 232 cases (84%) and 42 patients died. Prognosis factors identified were age > 65 years, Glasgow coma score < 7, convulsions, cardio-vascular shock, macroscopic haemoglobinuria. HIV infection has not been identified as a pejorative factor Our results confirm that severe malaria in native adult is a reality in tropical area. This study shows how difficult it is to have an adequate care management regarding this pathology in our context. PMID- 15787270 TI - [Serological survey of acquired toxoplasmosis in women of child-bearing age in Yopougon (Abidjan, Cote d'Ivoire)]. AB - The seroprevalence of toxoplasmosis was assessed from August 2000 to March 2001 on 1025 women in child-bearing age while consulting in the preventive health center (PHC) of Yopougon (Abidjan). Indirect immunofluorescent test was used for detection of IgG and Remington test for detection of IgM. Among the women under investigation, 60% were found to be IG seropositive, including 0.4% IgM seropositive. Soil and food were sources of contamination. When compared to previous studies conducted in the same region for several years, these results give evidence of a stability of the seroprevalence of toxoplasmosis, contributing to increase the risk of toxoplasmic encephalitis occurrence in AIDS patients. PMID- 15787271 TI - [Monitoring and assessment indicators in 2001 of "Roll Back Malaria" initiative in Benin]. AB - Within the context of WHO/CDS/RBM/2000, a survey was conducted in 2001 by the National Malaria Control Program of Benin. Following a well-thought-out choice, the survey took place simultaneously in health areas corresponding to epidemiological regions. Morbidity due to malaria is very high among children under five years admitted in external clinic (44.3% of cases) and (46.5%) for hospitalization. The crude rate of mortality is 129%. The use of non-impregnated bednet is usually met in three health areas, where 47.4% of the household have at least one non-impregnated bednet versus 5.4% of household with impregnated bednets. Percentage of pregnant women sleeping under an impregnated bednet and following chemoprophylaxis is respectively 43.3% and 3.8%. Results obtained at the end of this database survey in 2001 have facilitated the definition of indicators of the process, results and impact which remain very useful for the implementation of the monitoring and assessment system of "Roll Back Malaria" in Benin. PMID- 15787272 TI - [The 2003 International E. Brumpt Prize]. PMID- 15787273 TI - [The 2004 International E. Brumpt Prize]. PMID- 15787274 TI - Novel human neuropeptide Y Y5 receptor antagonists for the treatment of obesity. Synthesis and biological evaluation of pyridine hydrazide derivatives. AB - A series of new pyridine hydrazide derivatives with high and selective antagonist activity at the human neuropeptide Y Y5 receptor were developed. Introduction of electron-withdrawing groups into the arylsulfonamide rest, together with the 3 pyridyl analogue in the hydrazide moiety, led to a significant improvement of potency and solubility, affording trans-N-(4-[N'-(pyridine-3-carbonyl)hydrazino carbonyl]cyclohexylmethyl)-2,4-dichloro-benzenesulfonamide (14), which binds to the hY5 receptor with an IC50 value of 7.44 nmol/L. PMID- 15787275 TI - Restoration of endothelial function via enhanced nitric oxide synthesis after long-term treatment of raloxifene in adult hypertensive rats. AB - Raloxifene (CAS 84449-90-1, RAL), a selective estrogen receptor modulator (SERM) effective for the prevention of post-menopausal osteoporosis, also has been shown to acutely stimulate nitric oxide (NO) synthesis associated with improved endothelium-dependent relaxation. The effect of a 3-month RAL treatment (10 mg/kg/d) on basal blood pressure, measured via the carotid artery, and its challenge with increasing doses of intravenous bradykinin (1, 3 and 10 nmol/kg) was investigated. Furthermore, aortic NO bioavailability and relaxation in 9 month-old male and female ovarectomized (OVX) spontaneously hypertensive rats (SHR) was tested. Calcium ionophore stimulated NO release from aortic endothelial cells and aortic superoxide (O2-) production was directly assessed by using electrochemical nanosensors. Relaxation studies were performed with acetylcholine (10(-8) to 10(-5) mol/L) following precontraction with phenylephrine (10(-7) mmol/L). Whereas basal blood pressure (BP) was not significantly decreased in RAL treated SHR, the dose-dependent challenge with bradykinin induced an enhanced BP reduction in either gender. In contrast to female animals, aortic segments from RAL treated male animals showed significantly improved relaxaSHR) to 360 +/- 15 nmol/L. Vice versa, O2- was decreased from 110 +/- 15 to 22 +/- 1 nmol/L. In female SHR, ovarectomy led to an increase/decrease of NO/O2- from 130 +/- 5 to 180 +/- 10 nmol/L and 82 +/- 7 to 68 +/- 3 nmol/L, respectively. These effects were significantly amplified by RAL treatment (NO: 370 +/- 10 and O2-: 25 +/- 2 nmol/L). The results show that long-term treatment with RAL has beneficial affects on the cardiovascular system in old male and female OVX SHR via an increased NO bioavailability. PMID- 15787276 TI - In vitro and in vivo equivalence studies of alendronate monosodium tablets. AB - OBJECTIVE: The aim of this study was to test the bioequivalence of two alendronate tablets (CAS 121268-17-5; Marvil 10 and Marvil 70 as test formulations, in short "test"; reference formulation, in short "reference") in vitro and in vivo in healthy adult male subjects and to describe a mode for researching the bisphosphonate oral formulation pharmaceutical quality. METHODS: Two dissolution tests with 10-mg and 70-mg alendronate tablets, a preliminary clinical test with 10-mg tablets (n = 10) and a bioequivalence study with 70-mg tablets (n = 23) were performed. Clinical studies were single-dose, open, cross over, randomized, including a four-week wash-out period. Alendronate was assessed by HPLC in urine after 6 (UE6) and 24 (UE24) h post-intake. In all the experiments the reference was the one that had proved efficacy and safety in international regulatory clinical trials. RESULTS: The dissolution test showed a comparable release profile between reference and test, of both, the 10-mg and 70 mg tablet, the difference (f1) and similarity (f2) factors being within the acceptance values. The clinical trials showed great variability in urinary recovery, from one third the average figure up to 2-3 fold. The amount recovered with the 70-mg tablet was 11-15 fold higher than with the 10-mg tablets, suggesting higher (test/reference) was found to be 72-122% for UE24, and when analyzed in individuals with apparent steady bone metabolism during the wash-out period (n = 19), it was 86-137%. Both margins are considered acceptable in view of the particular kinetic and dynamic features of bisphosphonates, their very high inter- and intra-individual variability, extremely low absorption, time changeable bone compartment, high margin of safety and long-term achievable therapeutic benefits. CONCLUSION: Test is bioequivalent to reference. PMID- 15787277 TI - Synthesis and biological activity of some new flavone derivatives. AB - In this study, a new series of 2-(4-[substituted benzylamino-methyl)-phenyl]-4H benzopyrane-4-one (IVa-e) and N-substituted benzyl-N-[4-(4-oxo-4H-benzopyrane-2 yl)benzyl]-3-phenyl-acrylamide (Va-e) derivatives was synthesized and the results of their biological activity are reported. The synthesized compounds were tested for their in vitro antifungal and antibacterial activities. Compound IVa showed the best antifungal activity compared with miconazole (CAS 22916-47-8). Compound IVe indicated the same antibacterial activity compared with the control drug ampicillin (CAS 69-53-4). PMID- 15787278 TI - Synthesis and evaluation of the antibacterial and antitubercular activity of some N1-aral-N4-(3-chloro-4-fluorophenyl)thiosemicarbazones and their copper(I) complexes. AB - New N1-aral-N4- (3-chloro-4-fluorophenyl)thiosemicarbazones (2a-h) and their Cu(I) complexes (3a-h) have been prepared and characterized by elemental analysis and spectral methods (IR and 1H-NMR). Thiosemicarbazones 2a-h bind to copper(I) as bidentate ligand via nitrogen and thione sulphur centers to afford 3a-h. Both ligands and their complexes are screened against Escherichia coli (National Collection of Type Culture 10418) and Staphylococcus aureus (National Collection of Type Culture 5571) to evaluate their antibacterial properties and against human virulent H37Rv strain of Mycobacterium tuberculosis for their antitubercular properties. Ligands 2d-h showed promising antibacterial activity while their Cu(I) complexes 3d-h exhibited excellent activity. The compounds 2a, 2d, 2f, 2g and their Cu(I) complexes 3a, 3d, 3f and 3g were evaluated for their antitubercular activity. It was observed that compounds 2a, 2g, 3a, 3d showed moderate activity while 3f and 3g exhibited promising antitubercular activity. PMID- 15787279 TI - Anti-proliferative and anti-leukemic activity of DDE46 (compound WHI-07), a novel bromomethoxylated arylphosphate derivative of zidovudine, and related compounds. Studies using human acute lymphoblastic leukemia cells and the zebrafish model. AB - The anti-proliferative effects of a novel bromomethoxylated arylphosphate derivative of zidovudine (compound DDE46, CAS 213982-96-8) were first examined in a zebra fish embryo model. DDE46 blocked the cell division at the 2-cell stage of the embryonic development followed by total cell fusion. DDE46 also inhibited the proliferation of the leukemic cell lines NALM-6 and MOLT-3. DDE46 enhanced the activity of the pro-apoptotic enzymes Caspase-3, Caspase-6, Caspase-8, and Caspase-9 leading to the apoptotic death of the leukemic cell line Jurkat. These results justify the further development of this agent as a new anti-leukemic drug candidate. PMID- 15787280 TI - In vitro antifungal properties structure-activity relationships and studies on the mode of action of N-phenyl, N-aryl, N-phenylalkyl maleimides and related compounds. AB - The synthesis, in vitro antifungal evaluation and structure-activity relationship studies of 14 compounds of the N-phenyl-, N-aryl-, N-phenylalkyl- maleimide and 3,4-dichloromaleimide series are reported. The compounds were evaluated against a panel of standardized yeasts and filamentous fungi as well as clinical isolates of Candida albicans. The activities of N-phenylalkyl-3,4-dichloromaleimide derivatives but not those of N-phenylalkyl-maleimide derivatives showed to be dependent on the length of the alkyl chain. N-Phenylpropyl-3,4-dichloromaleimide showed the broadest spectrum of action and lower minimal inhibitory concentrations (MIC) in all of the fungi tested. The nitrogen-carbon distance between the two rings seems to play an important role in the antifungal behavior of these compounds. The most active structure showed inhibited (1,3)beta-D-glucan and chitin synthases, enzymes that catalyze the synthesis of the major fungal cell-wall polymers. PMID- 15787281 TI - [Is early debriefing useful or harmful?]. PMID- 15787282 TI - [Is monthly menstruation necessary?]. PMID- 15787283 TI - [Gefitinib therapy in non-small-cell lung cancer]. PMID- 15787284 TI - [Gefitinib as targeted treatment for non-small-cell lung cancer]. PMID- 15787285 TI - [Will the mystery of Parkinson's disease be resolved?]. PMID- 15787286 TI - [Oral care in connection with hematological stem cell transfer]. PMID- 15787287 TI - [Rodent dissemination of diseases in Finland]. PMID- 15787289 TI - [Central cyanosis and persisting eustachian valve]. PMID- 15787288 TI - [Lepidopterism. allergies and other reactions caused by butterflies]. PMID- 15787290 TI - [Breast cancer treatment]. PMID- 15787291 TI - [Surgery resolved malabsorption]. PMID- 15787292 TI - [Genetics of Alzheimer disease]. PMID- 15787293 TI - [Consequences of radiation]. PMID- 15787294 TI - [You too my Brutus]. PMID- 15787295 TI - [Comment to above:You too my Brutus]. PMID- 15787296 TI - [Meningococcal sepsis in a 15-year old girl's knee]. PMID- 15787297 TI - [Form a chain of survival for the management of cardiac arrest outside hospital!]. PMID- 15787298 TI - [Cardiac arrest outside hospital: initial and subsequent cardiological management]. AB - The prognosis of cardiac arrest outside hospital is directly related to the initial management. The aim of this work was to evaluate the characteristics of the initial and subsequent cardiological management of victims of cardiac arrest outside hospital with a retrospective analysis of data from the SAMU 35 (Emergency Medical Service, IIIe et Vilaine region) in the period April 1998 - April 2002. RESULTS: 533 non-traumatic cardiac arrests outside hospital were reported in 532 patients (average age 63 +/- 17, 73% male). The cardiac arrest occurred at home in 77% of cases. The initial cardiac rhythm documented was asystole in 63% of cases, ventricular fibrillation (VF) in 30% ventricular tachycardia (VT) in 1% and electromechanical dissociation in 6%. A cardiac aetiology was presumed in 294 (69%) of the 424 resuscitated patients. Among these, 22% (66/294) were admitted to coronary care units, 11% (31) left hospital alive, 8% (24) with no neurological sequelae. The survival rate for patients with cardiac arrest outside hospital in the presence of a witness and for whom the initial rhythm was VF or VT was 21%. The patient's age (<60 years)[OR: 1.05; CI 95%: 1.02-1.07; p < 0.001], rapid arrival of the SAMU (<10 min) [OR: 5.68; CI 95%: 1.42-22.7; p = 0.01] and resuscitation by the witness (OR: 8.26; CI 95%: 3.28-20.83; p < 0.001) were factors predictive of survival in a multivariate analysis. Coronary heart disease remains the principal cause of cardiac arrest in patients admitted to cardiology units (68%), with a recent coronary thrombosis shown in 40% of patients undergoing angiography (16/40). CONCLUSION: the prognosis of cardiac arrest outside hospital remains bleak, with a mortality of 90%. The survival rate is higher if the initial management is optimal, associated with bystander resuscitation and an immediate emergency service response allowing rapid defibrillation. Diagnosis and management of acute coronary syndrome in a cardiological setting must be integrated into the strategy. PMID- 15787299 TI - [Influence of sex on the prognosis of high risk acute coronary syndromes treated by early angioplasty]. AB - The aim of this study was to evaluate the influence of sex on the prognosis of high risk acute coronary syndromes treated early with angioplasty. Over a period of two years, 694 consecutive patients (151 female, 543 male) underwent revascularisation within the first 24 hours of an acute coronary syndrome without permanent ST elevation (ST depression (52.5%) or relapse of angina despite medical treatment (47.5%). The females were older than the males (67.9 vs 62.3 years; p < 0.0001), smoked less (7.3 vs. 32.8%; p < 0.001) and had a higher prevalence of hypertension (53 vs. 42.1%; p = 0.017). The angiographic characteristics were equivalent in both sexes, except for a lower frequency of thrombus in the females (6.9 vs. 15.2%; p < 0.0001). All lesions were treated with endoprosthesis implantation. The angiographic success rate was comparable (94 vs. 93.7%) as was the rate of major cardiac events while in hospital (3.8 versus 4%). With an average survival of 2 years, the incidence of major cardiac events remained identical in both sexes (15.4 vs 15.7%: p = 0.43): cardiac mortality (3.2 vs 2%; p = 0.18), myocardial infarction (7.3 vs 6.7%; p = 0.37), further revascularisation (8.3 vs 7.2%; p = 0.47). The survival without major cardiac event was comparable at 1 year (87 +/- 0.1 vs 88 +/- 0.3%) and at 2 years (78 +/- 0.2 vs 83 +/- 0.3%; p = 0.58). In conclusion, the progression both in hospital and at two years with a strategy of early revascularisation for high risk acute coronary syndromes was comparable in males and females. PMID- 15787300 TI - [Value of systematic biological markers of inflammation for the prognosis at 12 months of patients undergoing programmed coronary angioplasty]. AB - Value of systematic dosage of biological markers of inflammation for the prognosis at 12 months of patients undergoing programmed coronary angioplasty Systematic dosage of proteins of inflammation has been suggested for assessing the prognosis of athero-thrombotic diseases. The authors undertook a study of plasma C-reactive protein (CRP) and interleukin 6 (IL-6) for evaluating the prognosis of patients undergoing programmed coronary angioplasty. A prospective monocentric study of 117 patients (65 +/- 8 years) was divided into a control group of 28 patients undergoing coronary angiography (Group 1) and 89 patients undergoing programmed coronary angioplasty (Group 2). Serum IL-6 and CRP levels were measured before arterial puncture and at H12 and H24 after coronary catheterisation. The follow-up period was 12 months. The angioplasty did not significantly increase CRP and IL-6 concentrations compared with coronary angiography. Twenty patients (Group 2) (22%) suffered a cardiovascular event in the 12 months' follow-up. These patients had significantly higher CRP levels at H0, H12 and H24 after coronary angioplasty than those who had uncomplicated outcomes. This was not observed for IL-6 concentrations because of the wide dispersion of the results obtained. Increased CRP concentrations between H0 and H24 was also a good predictive factor independently of high basal CRP levels potentially due to other causes than atheroma. Coronary angioplasty is associated with increased CRP at H0, H12 and H24. These values are correlated with the risk of future events at 6 and 12 months. This information is easily obtained and should help management of these patients. PMID- 15787301 TI - [Survey of cardiovascular risk assessment and the behaviour of a sporting population]. AB - In order to better understand the risk factors and behaviour of a general sports population, a questionnaire was handed to 603 consecutive sportsmen of various disciplines during a specific sports medicine consultation and a half-marathon. Among the discovered risk factors, smoking was the most common (19.3% for the entire study population), predominantly in the youngest group (26%). A resting ECG had been performed very frequently within the previous year in those over 40 years (86.5%), but less often in the younger sportsmen (44.8%). An exercise test had also very often been performed in those aged over 40 years (69.2 vs 10.6%). Behaviour associated with cardiovascular risk was common. Undertaking physical activity while febrile (58%) and smoking just before or after exertion (13.1%) were widespread behaviours. This population appeared casual in its approach to possibly suspicious cardiac symptoms of exercise, particularly the females and paradoxically the subjects doing the most sport. Lastly, only a minority of sportsmen knew the telephone number for the emergency medical service (45%) or claimed to be competent at first aid (32%). This survey underlines a practice relatively in accordance with the recommendations for detecting cardiovascular pathology, but shows a failure of preventive education. PMID- 15787302 TI - The Brugada Syndrome. AB - In 1992 a syndrome was described consisting of syncopal episodes and/or (resuscitated) sudden death in patients with a structurally normal heart and a characte ristic electrocardiogram (ECG) displaying a pattern resembling a right bundle branch block with ST segment elevation in leads V1 to V3. The disease is genetically determined with an autosomal dominant pattern of transmission in 50% of the familial cases. Several different mutations have been identified affecting the structure, function and trafficking of the sodium channel. The syndrome is ubiquitous. Its incidence and prevalence are difficult to estimate, but this disease may cause 4 to 10 sudden deaths per 10,000 inhabitants per year representing the most frequent cause of natural death in males younger than 50 in South Asia. The disease has been linked to the sudden infant death syndrome (SIDS) and to the sudden unexpected death syndrome (SUDS) by showing that the electrocardiogram and mutations are the same as in Brugada syndrome. The diagnosis is easily made by means of the ECG when it is typical. There exist, however, patients with concealed and intermittent electrocardiographic forms that make the diagnosis difficult. The ECG can be modulated by changes in autonomic balance, body temperature, glucose level and the administration of antiarrhythmic, neuroleptic and antimalaria drugs. Beta adrenergic stimulation normalizes the ECG. Loss of the action potential dome in right ventricular epicardium but not in endocardium underlies the ST segment elevation. Electrical heterogeneity within right ventricular epicardium leads to the development of closely coupled extrasystoles via phase 2 reentry that precipitate ventricular ,fibrillation. Antiarrhythmic drugs do not prevent sudden death in symptomatic or asymptomatic individuals. Implantation of an automatic cardioverter-defibrillator is the only currently proven effective therapy. Patients with frequent electrical storms may even need cardiac transplantation as last resort. PMID- 15787303 TI - How to accelerate the endothelialization of stents. AB - Coronary artery stenting is currently the most frequently performed percutaneous coronary intervention for the treatment of coronary artery disease. The endothelium is a single layer of endothelial cells lining the vascular wall and plays an integral part in maintaining vascular homeostasis. Stenting however causes significant injury to the vascular wall and endothelium, resulting in inflammation, repair and the development of neointimal hyperplasia. The ability of the endothelium to repair itself depends on both the migration of surrounding mature endothelial cells, and the attraction and adhesion of circulating endothelial progenitor cells (EPCs) to the injured region, which then differentiate into endothelial-like cells. Current therapies with drug-eluting stents interrupt the natural response to damage. Accelerating the reendothelialization of the damaged arterial segment following stent implantation is an attractive form of therapy as it is seen as hastening the natural process of repair. It potentially has the benefit of reducing the amount of neointimal hyperplasia and stent thrombosis. Studies have been performed to identify agents that augment the mobilisation and recruitment of EPCs to the injured area (statins, exercise, estrogen and cytokines). Other studies have looked at seeding stents with endothelial cells or EPCs. The most current approach is to coat anti CD34 antibodies on a stent surface to attract circulating EPCs to the stent which then differentiate into endothelial-like cells. This approach is currently being tested in safety and feasibility clinical studies. PMID- 15787304 TI - [Prevalence of dyslipidaemias in a representative sample of the French population]. AB - Prevalence of dyslipidaemias in a representative sample of the French population Hypercholesterolaemia is a major factor of risk of coronary atherosclerosis. The prevalence of other types of dyslipidaemia in the general population remains poorly defined. This study was performed to measure the prevalence of various dyslipidaemias in the French population. A representative sample of 3508 men and women between the ages of 35 and 64 years was recruited by the "Multinational MONItoring of trends and determinants in CArdiovascular disease" centres of Lille, Strasbourg and Toulouse. We excluded 162 patients suffering from known cardiovascular disorders, and 409 individuals treated with lipid-lowering drugs. The prevalence of pure hypercholesterolaemia, defined as a total cholesterol concentration >6.2 mmol/l (2.4 g/l) and triglyceride concentration <2.3 mmol/l (2 g/l), was 30% (29-32%). The prevalence of HDL cholesterol concentration <1 mmol/l (0.4 g/l) in men, or <1.3 mmol/l (0.5 g/l) in women, was 12% (11-13%). The prevalence of mixed hyperlipidaemia, defined as a total cholesterol concentration >6.2 mmol/l (2.4 g/l) and triglyceride concentration >2.3 mmol/l (2 g/l) was 5% (4-6%). The prevalence of hypertriglyceridaemia, defined as a total cholesterol concentration <6.2 mmol/l (2.4 g/l) and triglyceride concentration >2.3 mmol/l (2 g/l) was 4% (3-5%). Low HDL cholesterol concentrations were associated with smoking, obesity, and absence of either regular physical exercise or alcohol consumption. This study confirmed the high prevalence of pure hypercholesterolaemia, and revealed an important prevalence of low HDL cholesterol concentration, which represents a major cardiovascular risk factor. PMID- 15787305 TI - [Cognitive functions and hypertension]. AB - The prevention of cognitive disorders and dementia represents a major challenge in the coming years. Hypertension is one of the principal risk factors for cerebrovascular diseases and is also closely correlated with cognitive decline and dementia. Most longitudinal studies have shown that cognitive functioning is often inversely proportional to blood pressure values measured 15 or 20 years previously. The higher blood pressure was, the poorer cognitive function is. Data from recent therapeutic trials (SYST-EUR, PROGRESS) open the way toward the prevention of dementia (vascular or Alzheimer's type) by antihypertensive treatments. In this context, the effect of antihypertensive treatment on cognitive functions should represent one of the primary criteria of assessment in future morbidity and mortality studies in hypertensive patients. PMID- 15787306 TI - [Role of antiarrhythmic drugs in reducing the number of defibrillation shocks]. AB - There is a logical basis for the prescription of antiarrhythmic therapy (AAT) in patients with an implanted automatic defibrillator (IAD): the prevention of supraventricular and ventricular arrhythmias, slowing the rate of a ventricular arrhythmia to improve the haemodynamic tolerance and better efficacy of anti tachycardia stimulation. Adjuvant therapy of this kind is used in 49 to 69% of cases. Forty six per cent of patients in the MADIT I trial had AAT at the last follow-up. In the AVID trial, despite recommendations to avoid the association of AAT and [AD, 18% of patients had this combination because of the large number of shocks in 64% of cases; AAT was associated with both a reduction in the number of shocks (p < 0.05) and in the delay to a recurrence of the arrhythmia (p < 0.0001). In the CASCADE trial, carried out in survivors of cardiac arrest, the prescription of amiodarone in patients with IAD significantly reduced the number of defibrillation shocks. Finally, almost 40% of patients with IAD have an "arrhythmia storm", sometimes during follow-up, usually requiring the association of AAT. Despite the frequent association of AAT and IAD, the results of this combination have rarely been studied systematically. Only one randomised trial has been published comparing D,L-sotalol and placebo in patients with IAD. The treatment reduced the probability of a first defibrillation shock, whether appropriate or not. Amiodarone has often been proposed in association with IAD in isolated cases. The OPTIC trial (Optimal Pharmacological Therapy in Implantable Cardioverter), currently under way, has three sub-groups of IAD (betablocker, amiodarone and betablocker, sotalol), with the aim of determining the preventive value of each AAT with respect to the IAD performance. In addition to the results of combined treatment, the interaction with the threshold of defibrillation is an important factor which is not clearly understood at present. Class I AAT increases the threshold of defibrillation whereas Sotalol reduced it. Amiodarone has a variable effect, neutral or deleterious. The OPTIC trial should provide valuable information via the sub-analysis of the effects of drugs on the threshold of defibrillation in each of the three arms of the trial. PMID- 15787307 TI - [Homocysteine, folic acid, group B vitamins and cardiovascular risk]. AB - Although there is a high degree of proof relating plasma homocysteine levels to cardiovascular risk, the role of homocysteine as a causal cardiovascular risk factor remains controversial. Prospective long-term clinical trials in high cardiovascular risk populations usually show a positive relationship between plasma homocysteine and the degree of cardiovascular risk. However, shorter term studies and/or those carried out in populations with lower cardiovascular risk show either a weaker correlation or no relationship at all. To date no study has shown proof of the reversibility of cardiovascular risk due to hyperhomocysteinaemia; nevertheless, a number of studies using intermediate criteria support the hypothesis of a benefit due to reduction of plasma homocysteine levels. A number of therapeutic trials published with clinical criteria have not shown convincing results in either direction. A number of interventional trials are underway: notably the SUFOLOM 3 trial in France, and the question of a benefit on cardiovascular risk by reducing homocysteine levels should be answered in the next few years. In the meantime, with the exception of homocysteinuria in which therapeutic strategies have shown their efficacy in the reduction of atherothrombotic risk with high levels of proof, the authors do not recommend the treatment of mild hyperhomocysteinaemia in any clinical setting other than "clinical trials" and certain "compassionate" indications such as early and/or recurrent vascular events associated with hyperhomocysteinaemia in the absence of conventional risk factors. PMID- 15787308 TI - [Intracerebral haematoma during a stress test]. AB - We report the case of a 64 year-old moderately hypertensive patient investigated for dyspnoea on exertion with no chest pain. After informing the patient, an ergometric test following a gentle protocol was performed, according to the French Society of Cardiology guidelines. At the peak of effort the patient developed a sudden left hemiplegia with a right capsulo-thalamic haematoma on cranial CT. No other case has been described and a literature search showed no relationship between physical effort and cerebral haematoma. Following an administrative tribunal enquiry, no medical fault was attributed regarding the indication and performing the test; no failure to inform could be established for a risk that was unknown at the time of the test. PMID- 15787309 TI - [Torsades de pointes caused by cetirizine overdose]. AB - Several therapeutic substances can cause torsades de pointes, especially if they prolong the QT interval and/or if there is an associated hypokalaemia. Certain second generation H1 antihistamines have been incriminated in the occurrence of such ventricular arrhythmias, such as terfenadine and astezimole, which have been withdrawn. Cetirizine, widely used in the treatment of allergic reactions, is a second generation H1 antihistamine with as yet no precautions of use regarding rhythm disturbances. No documented case of arrhythmia attributable to this drug has been reported. We report the case of a dialysed patient with chronic renal failure who had symptomatic episodes of torsades de pointes in the context of hypokalaemia and cetirizine overdose. In the light of this observation it would appear that the prescription of cetirizine is contra-indicated under such conditions. PMID- 15787310 TI - [Infectious endocarditis in cerebral death: implications for explantation of tissues and organs]. AB - Allograft valve replacement is the preferred treatment for infectious endocarditis With severe annular destruction. Explantation of the valve in patients with cerebral death requires preliminary cardiac investigations and strict surgical procedures. The authors report two cases of right atrial endocarditis discovered at the time of explantation of the valvular allografts. In view of the infectious nature of the lesions, no tissues were resected. In both cases, transthoracic echocardiography had not diagnosed these lesions. The sensitivity of transoesophageal echocardiography in the diagnosis of these lesions should lead to systematic referral for this investigation. The infectious nature of these endocarditic lesions should raise suspicion about the presence of a central venous catheter. In addition, careful inspection of the chambers, septa and cardiac valves should be performed at surgery. The detection of potentially infectious endocarditic lesions should, in accordance with present recommendations, lead to renouncing the explantation of valvular allografts. PMID- 15787311 TI - [Collaboration between radiologists and cardiologists]. PMID- 15787312 TI - [Pilot study of echocardiographic studies using color- and pulsed-wave spectral Doppler methods in blue-crowned amazons (Amazona ventralis) and blue-fronted amazons (Amazona a. aestiva)]. AB - Colour-flow and pulsed-wave spectral Doppler echocardiography was performed on 6 healthy, adult Hispaniolan amazon parrots (Amazona ventralis) and 6 blue-fronted amazon parrots (Amazona a. aestiva) to establish normal reference values. Birds were anesthetized with isoflurane in oxygen and placed in dorsal recumbency. An electrocardiogram was recorded continuously and birds were imaged with a micro phased-array scanner with a frequency of 7.0 MHz. After assessment of cardiac function in 2-D-echocardiography, blood flow across the left and the right atrioventricular valve and across the aortic valve was determined using color flow and pulsed-wave spectral Doppler echocardiography. Diastolic inflow (mean value +/- standard deviation) into the left ventricle was 0.17 +/- 0.02 m/s (Hispaniolan amazons) and 0.18 +/- 0.03 m/s (Blue fronted amazons). Diastolic inflow into the right ventricle was 0.22 +/- 0.05 m/s (Hispaniolan amazons) and 0.22 +/- 0.04 m/s (Blue fronted amazons). Velocity across the aortic valve was 0.84 +/- 0.07 m/s (Hispaniolan amazons) and 0.83 +/- 0.08 m/s (Blue fronted amazons). Systolic pulmonary flow could not be detected in any of the birds in this study. No significant differences were evident between the two species examined. Results of this study indicate that Doppler echocardiography is a promising technique to determine blood flow in the avian heart. Further studies in other avian species are needed to establish reference values for assessment of cardiac function in diseased birds. PMID- 15787313 TI - [Development of cell content and shedding of Prototheca spp. in milk from infected udder quarters of cows]. AB - It was the objective of this study to analyse shedding patterns and somatic cell counts in cows and quarters infected with Prototheca spp. and to evaluate two approaches to identify infected animals by somatic cell count (SCC) or by bacteriological analysis of pooled milk samples. Five lactating dairy cows, chronically infected with Prototheca spp. in at least one quarter were studied over 11 weeks to 13 months. Quarter milk samples and a pooled milk sample from 4 quarters were collected aseptically from all quarters of the cows on a weekly basis. Culture results of quarter milk and pooled samples were compared using cross tabulation. SCC of quarter milk samples and of pooled samples were related to the probability of detection in the infected quarters and cows, respectively. Shedding of Prototheca spp. was continuous in 2 of 8 quarters. In the other quarters negative samples were obtained sporadically or over a longer period (1 quarter). Overall, Prototheca spp. were isolated from 83.6% of quarter milk samples and 77.0% of pooled milk samples of infected quarters and cows. Somatic cell counts were higher in those samples from infected quarters that contained the algae than in negative samples (p < 0.0001). The same applied for composite samples from infected cows. Positive samples had higher SCC than negative samples. However, Prototheca spp. were also isolated from quarter milk and pooled samples with physiological SCC (i.e. < 10(5)/ml). Infected quarters that were dried off did not develop acute mastitis. However, drying off had no effect on the infection, i.e. samples collected at calving or 8 weeks after dry off still contained Prototheca spp. Results indicate that pre-selection of cows to be sampled for Prototheca spp. by SCC and the use of composite samples are probably inadequate in attempts to eradicate the disease. However, due to intermittent shedding of the algae in some cows, single herd sampling using quarter milk samples probably also fails to detect all infected cases. Therefore, continuous monitoring of problem cows with clinical mastitis or increased SCC in herds during eradication programs is recommended. PMID- 15787315 TI - [Survival analysis of German Holstein cows after an abomasal displacement]. AB - From 1996 to 2002 5159 cattle with abomasal displacement was treated in the Clinic for Cattle, School of Veterinary Medicine Hannover. An analysis of systematic effects on influences on the length of survival following surgical correction of the abomasal displacement could be performed for 1411 cows. These animals were under the official milk recording scheme. Significant influences on lifetime after an abomasal displacement were interval between calving and diagnosis of disease, lactation number, type of abomasal displacement, inbreeding coefficient, calving difficulties, region of origin and sire of the calf. The lifetime after an abomasal displacement significantly differed between several months of calving and some sires of the cow. The individual milk yield in lactations before and after abomasal displacement of the cow was significant for the length of lifetime. The heritability estimate for length of lifetime after abomasal displacement using residual maximum likelihood (REML) in a linear animal model was h2 = 0.01-0.03. PMID- 15787314 TI - [Precision-controlled echocardiographic left ventricular function parameters by repeated measurement on three consecutive days in trained and untrained warmblood horses]. AB - Echocardiographic measurements have to be reliable and reproducible with only a low day to day variability for detecting pathological changes of left ventricular myocardial function in horses. The day dependent fluctuation margin represents an important indicator for the reproducibility of a method. To find out the day to day variability of echocardiographic parameter in healthy warmblood horses, in the present study repeated echocardiographic measurements at three consecutive days were carried out (at the same time and by the same examiner) at 11 untrained and 7 trained warmblood horses. The horses were examined from the right hemithorax, in five different B- and M-Mode views in the long and short heart axis. The echocardiographic measurements in the three-day examination interval showed with coefficient of variation between 3.4 and 25.8% a good reproducibility. In order to calculate the precision of echocardiographic parameters (reproducibility from day to day) statistical analysis was carried out by means of a linear model with random effects. Herefor the variation between repeated measurements over three days and within the horses (day to day intraindividual variation) as well as the variation between the horses (interindividual variation) were taken into account. In dependency of the echocardiographic parameters 46.2-95.7% (median 85.7%) of the variation is caused by the difference between the horses (intraclass correlation) and 4.3-53.8% (median 14.3%) of the variation is caused by the repeated measurements on different days. With a coefficient of variance from 6.6% and an intraclass correlation of 0.957 the left ventricular diastolic diameter at the level of the papillary muscles showed the lowest variation between repeated measurements on different days. The heart rate with a coefficient of variance of 10.3% and an intraclass correlation of 0.462 showed the highest variation with 53.8% between the repeated measurements on different days. The precision of single echocardiographic measurements is confirmed. All parameters were also tested for differences between trained and untrained horses. Significant differences (p<0.05) could be found in the left ventricular diameter at the level of the apex cordis and under the mitral valves as well as in the systolic left ventricular area. PMID- 15787316 TI - [Saponins in nutrition of swine, poultry and ruminants]. AB - Saponins are glycosides found in many plants, which have soap character due to their surfactant properties. They work haemolytic and may cause symptoms of intoxication in high concentration. Among therapeutically relevant effects expectorative, anti-inflammatory and immune-stimulating effects have the highest ranking. Beyond that, saponins demonstrate antimicrobial properties particularly against fungi and additionally against bacteria and protozoa. In animal nutrition additions of saponins can suppress intestinal and ruminal ammonia production. The ammonia-reducing effect is primarily attributed to an inhibition of proteolytic microorganisms. Due to reduced NH3-concentrations metabolism is relieved, which offers benefits to animal welfare and animal performance. Accordingly improved feed convertion ratio can be registered in weanling and growing-finishing pigs with the employment of saponin-containing feed additives. With regard to ruminal fermentation in vitro and in vivo studies have shown that beside proteolytic protozoa also cellolytic and amylolytic bacterial species are restrained. To what extent antimicrobial activity supress digestion or whether the selective effects on microorganisms can be used purposefully to manipulate ruminal fermentation, must be clarified in further investigations. PMID- 15787317 TI - [Focused extracorporeal shock wave therapy in an isolated calcification in the equine ligamentum nuchae--case report]. AB - In recent years extracorporeal shock wave therapy (ESWT) is used in veterinary medicine especially in equine orthopedics. Different ESWT generators are commonly used in equine medicine with varying energy density and depth of penetration. The presented case report described the treatment of a ligamentum nuchae calcification with focused extracorporeal shock wave therapy with SONOCUR Plus generator combined with SONOLINE Prima ultrasound. Negligible adverse effects, the non-invasive character of the treatment, the time-saving therapy, and the positive results make the focused extracorporeal shock wave therapy to an useful application. PMID- 15787318 TI - Occlusion, bruxism, and dental implants: diagnosis and treatment for success. PMID- 15787319 TI - Coincidental adrenal ganglioneuroma--a case presenting with enuresis nocturna. AB - A 9-year-old boy presenting with primary enuresis nocturna was discovered to harbour a large left adrenal mass which produced no hormonal abnormalities. He was undergone an exploration and the mass was removed completely. It was found to be an adrenal ganglioneuroma weighing 240 g. Though no cause-and-effect relationship is established, the case no long suffered from enuresis soon after surgery and free of recurrence after 5 years. PMID- 15787320 TI - A case report: adrenal ganglioneuroblastoma in a 59-year old man. AB - Ganglioneuroblastoma is a common type of tumor in children, but is rarely seen in adults. We present here a case of adrenal ganglioneuroblastoma in a 59-year old man. PMID- 15787321 TI - Renal sinus hemangioma simulating renal mass: a diagnostic challenge. AB - A 47-year-old female presenting with right flank pain and renal mass on ultrasonography was evaluated for renal malignancy. Based on the CT findings and blood aspiration on repeated fine needle aspiration biopsy the patient underwent radical nephrectomy. Histopathology revealed renal sinus hemangioma with normal kidney, perirenal fat and the hilar structures. PMID- 15787322 TI - Xanthogranulamatous pyelonephritis with psoas abscess: 2 cases and review of the literature. AB - Xanthogranulomatous pyelonephritis (XGP) is a rare form of chronic pyelonephritis. With the review of the literature, we reported two cases of XPN with psoas muscle abscesses caused by Staphylococci aereus in one of the patient and Serratia mascerentes in the other. Both of the patients had renal calculus. We performed nephrectomy with psoas abscess drainage and started appropriate antibiotics, but one of the patients died of septic shock. Other patient is free of symptoms at the end of 5 years follow-up. PMID- 15787323 TI - Idiopathic fibrosclerosis of bilateral orbits, bilateral ureters, thyroid: a case report and review of the literature. AB - INTRODUCTION: Idiopathic multifocal fibrosclerosis is a rare disorder which is usually misdiagnosed in the clinics. We try to make a review of the literature about this large spectrum syndrome and treatment modalities other than surgery. Our patient interestingly have both fibrosclerosis of bilateral ureters, orbits and thyroid. Steroids, radiotherapy, tamoxifen, vitaminD3, colchisine was found to be in effective in treatment of progressive fibrosis. PMID- 15787324 TI - Synchronous ipsilateral conventional renal cell and transitional cell carcinoma. AB - Simultaneous occurrence of renal cell carcinoma (RCC) and transitional cell carcinoma (TCC) in the same kidney is unusual. We report a 61-year-old man with ipsilateral synchronous renal adenocarcinoma and renal pelvic TCC. He was referred to our department for gross hematuria and right flank pain. CT and MRI studies revealed a 57 x 50 mm irregular and infiltrative upper right kidney mass with necrotic components. A right radical nephrectomy was done. Pathological diagnosis was a high grade tumor originating from just beneath the intact urothelium of renal pelvis and infiltrating through the parenchyma showing solid and occasional tubular growth patterns. A second tumor in close proximity to the first was reported as well differentiated RCC. This is a rare case of combined renal malignancies. PMID- 15787325 TI - Routine ureteral dilatation is not necessary for ureteroscopy. AB - PURPOSE: To present our experience in ureteroscopic lithotripsy and stone extraction without ureteral dilatation. PATIENTS AND METHODS: A total of 134 consecutive patients (80 male and 54 female), with a mean age of 36.4 (18-65) years underwent ureteroscopic stone removal. The stones were located in the lower, middle, and upper parts of the ureter in 92, 18 and 24 patients and the mean stone diameters were 9.2 (6-15) mm, 10.5 (8-15) mm and 8.8 (8-10) mm, respectively. A semirigid ureteroscope 8 F in size was used without any ureteral dilatation. The stones were fragmented by a pneumatic lithotripter in the ureter and the fragments were removed by a basket catheter or stone forceps. All patients were re-evaluated with a plain film on postoperative first day and with intravenous urography (IVU) at 3 months. Residual fragments bigger than 3 mm were accepted as treatment failure. RESULTS: The mean operation time was 44 (20-120) minutes. After the operation, the stone-free rate was 89/92 (97%) for lower, 15/18 (83%) middle and 18/24 (75%) upper ureteral stones, respectively. Double J catheter replacement was needed in 13 patients due to impacted stone and/or failed procedure. Ureteral perforation did not occur in any patient. Patients were discharched from hospital within 6-24 hours. No ureteral stricture was encountered during the follow-up period. CONCLUSION: Our experience suggests that ureteroscopic interventions could be easily performed for all parts of ureter without previous dilatation of the ureter. PMID- 15787326 TI - Clinical efficacy of distigmine bromide in the treatment of patients with underactive detrusor. AB - PURPOSE: The aim of this study was to assess the clinical efficacy of distigmine bromide, an anticholinesterase agent, deemed to improve detrusor function thereby restoring normal voiding patterns in patients suffering from detrusor underactivity. MATERIALS AND METHODS: A total of 27 patients (11 men and 16 women) with poor detrusor function were included in the study. The diagnosis was established using pressure-flow studies. All patients received distigmine bromide at a dose of 5 mg three times daily for 4 weeks and re-attended for a follow-up urodynamic investigation. The results of baseline pressure-flow studies were compared to those after completion of treatment. RESULTS: Treatment with distigmine bromide resulted in a statistically significant reduction of residual volume and percent residual volume, obviating the need for intermittent self catheterisation in 11 patients. In addition, maximum flow rate and detrusor pressure at maximum flow increased, although not significantly. The drug was generally well tolerated by the majority of patients. CONCLUSION: Distigmine bromide shows clinical efficacy in patients with poor detrusor function and may therefore be used alternatively in selected cases. PMID- 15787327 TI - Nephrogenic adenoma of the urinary bladder. AB - INTRODUCTION: The nephrogenic adenoma (NA) is a benign metaplastic lesion of the urothelium and is attributed to chronic irritation of the mucosa, by injury, infection, stone disease or intravesical instrumentations. We present our experience on this morbid entity, its clinical appearance in the urinary bladder, its frequency and relapses. Furthermore we reviewed the related recent literature and focused on its potential to neoplastic degeneration and the value of the new diagnostic modalities. PATIENTS AND METHODS: Four patients with NA of the urinary bladder are presented. The papilloid or polypoid formations observed by the cystoscopy were identified after the TUR, as NA of the urinary bladder. Their mean follow-up was 3.5 years. RESULTS: Remission of the symptoms was observed after TUR in all patients. Three out of four patients presented 1-7 relapses, while in one case, after seven NA relapses, a urothelial carcinoma of the bladder was diagnosed. CONCLUSIONS: Unlike histological features, the clinical- endoscopic characteristics of NA are non-specific. Even if it is not definitely considered like a premalignant condition, NA has to be followed up frequently and long lasting, because of its high recurrence rate. The combination of Cytology, Flow cytometry, DNA image analysis and Fluorescence in situ hybridisation of bladder washings or voided urine, are of high value in monitoring NA of the urothelium. PMID- 15787328 TI - Incisional hernia: a possible complication after the orthotopic urinary diversion? AB - AIM: One of the complications of abdominal surgical procedures is incisional hernia. This complication is encountered in different rates due to the surgical technique, type of urinary diversion preferred, and whether the patient has additional risk factors or not. PATIENTS AND METHODS: In this study, 145 patients who had undergone urinary diversions in our clinic were evaluated between the years 1989 and 2002. Of those, 17 were treated by Mainz pouch type II urinary diversion, 47 by Indiana type urinary diversion and 81 patients were treated by orthotopic urinary diversions. RESULTS: Incisional hernia did not occur in any of the patients who had undergone Mainz pouch type II and Indiana type urinary diversions. Eleven of 145 patients (7.5%) who had undergone urinary diversions developed incisional hernia. All of these incisional hernia occurred in patients who had undergone orthotopic type urinary diversion. These incisional hernias occurred within the first postoperative year (2-8 months). CONCLUSIONS: We believe that increased intraabdominal pressure for micturition is the predisposing factor for the development of incisional hernias. Furthermore, the patients must avoid from sudden increase of intraabdominal pressure such as suddenly, strong valsalva maneuver during voiding. And we believe usefully press doing from the outside to abdomen during voiding (crede maneuver). PMID- 15787329 TI - What is the ratio of urethral recurrence risk after radical cystoprostatectomy for bladder cancer? AB - OBJECTIVE: There is always a risk of urethral recurrence after radical cystoprostatectomy in patients with bladder transitional cell cancer. Taking these risk factors of urethral recurrence into account, orthotopic neobladders or urinary diversions without using the urethra are performed. But urethral tumour recurrence occurs much less than the expected. We assessed the etiological factors that affect the urethral recurrence in orthotopic and nonorthotopic urinary diversion cases. METHODS: Sixty-four patients with bladder cancer who underwent radical cystoprostatectomy and urinary diversion between 1994 and 2002 were included this study. Conventional risk factors effecting the selection of operation type and urethral recurrence were evaluated in these patients. Cystoscopy and biopsy were done and pathologic specimen was obtained preoperatively, and cystoscopy and urethral washout cytology were done postoperatively. Routine bladder biopsies were done in uncertain cases at follow up. Risk factors increasing the urethral recurrence are as follows: papillary and multiple tumours, tumour invading bladder neck and trigone, extensive CIS, prostatic stromal and urethral invasion, positive surgical margin and history of upper urinary tract tumour. In 31 patients having one or more of these criteria, continent nonorthotopic urinary diversion was performed, but 33 patients without these risk factors underwent orthotopic urinary diversion. Simultaneous urethrectomy was not done in any of these patients. RESULTS: Among the patients who underwent radical cystoprostatectomy, none was with positive surgical margin in the distal end of the prostatic urethra. In preoperative cystoscopy, tumoural mass was seen near to collum in eight patients and in the prostatic urethra in three patients. Histopathological examination of cystoprostatectomy specimen displayed transient epithelial cell carcinoma of prostatic urethra in three patients, transient epithelial cell metaplasia inside the prostate in five patients and invasion to the urothelium of bladder neck in three patients. There were not any transient epithelial cell cancer metastases in prostatic stroma in any of these patients. One patient underwent urethrectomy, since atypical cells were observed in postradical prostatectomy urethral washout cytology but there was no tumour found in pathological examination of the specimen. Therefore, urethral tumour recurrence did not occur after 25 months follow up. CONCLUSION: These findings suggest that some of the conventional risk factors of urethral recurrence were exaggerated. We may also conclude that there is no need for prophylactic urethrectomy unless there is urethral cancer or cancer in the surgical margin. But if utilization of urethra is planned, evaluation of prostatic stroma by TUR biopsies and urethral anastomose margin by frozen section during the operation is necessary. PMID- 15787330 TI - Urothelial carcinoma of the bladder with trophoblastic differentiation: a case report. AB - We report a case of a urothelial carcinoma with trophoblastic differentiation of the bladder that occured in a 23-year-old man. The patient presented with hematuria. Multiple papillary masses were resected transurethrally. Histopathologically, grade III urothelial cell carcinoma contained giant cells that were positive for human chorionic gonadotrophin (HCG), placental alkalen phosphatase, and human placental lactogen. HCG secreting tumors are reported to be highly aggressive. The patient is alive and well without evidence of recurrent disease or metastasis at 10 months from transurethral resection. To our knowledge, this case is the youngest patient in the literature so far. PMID- 15787331 TI - Folding the flow rate: a simple method to display and file the uroflowmetry tracing. PMID- 15787332 TI - Suprapubic catheter knotting: an unusual complication. AB - We describe a case of true knotting of a suprapubic catheter in a 25-month-old boy who underwent surgery for urethrocutaneous fistula as a complication of a distal penile hypospadias repair. This unusual complication was probably attributable to an excessive length of catheter being inserted into the bladder, thereby allowing it to bend onto itself. PMID- 15787333 TI - Mass screening of prostate cancer in Changchun City of China. AB - OBJECTIVES: Mass screening of prostate cancer in China had not been studied until now. To clarify the incidence of prostate cancer among the population of aged men in Changchun City of China, we performed a mass screening program for prostate cancer using prostate specific antigen (PSA). METHODS: Men aged more than 50 years were examined by PSA, with a cut-off values 4.1 ng/ml. Subjects, whose serum PSA concentration was more than 4.1 ng/ml, were recommended to secondary examination of transrectal ultrasound-guided systematic six-sextant biopsy examination (TRUS-guided biopsy). RESULTS: Between April 1998 and December 2000, in the urban area of Changchun City 4218 men over 50 years old were screened. As 190 men were found to have elevated PSA and 116 men underwent a TRUS-guided biopsy of the prostate, of whom 29 men were found to have cancer. In all, the age adjusted PSA positive rate was 4.80%, cancer detection rate was 0.74%. The age adjusted cancer detection rate was 0.78%. After adjusted for secondary examination rate, the cancer detection rate reached 1.28%. The cases of stage B with limited cancer within the prostate gland, accounted for 62% of 29 cancer cases. Stage B was much more detected (accounted for 60%) in the PSA range of 4.1 10.0 ng/ml. Stage C and D were much more detected (accounted for 85.71%) in the PSA range over 10.1 ng/ml, with a significant difference by Fisher exact test (P < 0.05). A significant correlation of age-adjusted PSA positive rate and age adjusted cancer detection rate adjusted by secondary examination rate was showed in six groups of Community and Army, Factory, Corporation, Post and Telecom Office, Government and Academy, and University and Technical School (r = 0.898, P < 0.01). Both of the age-adjusted PSA positive rate and age-adjusted cancer detect rate of "Universities and Technical Schools" were significantly higher than that of "Factories" (chi2 test P < 0.05). CONCLUSIONS: The prevalence of prostate cancer was not so low in Chinese people as expected. For 50 years or older men, cancer detection rate was 0.74%, The age-adjusted cancer detect rate was 0.78%. Much more early stage cancer could be detected in the screening using serum PSA. For the different company the detection rate of prostate cancer varied each other among the groups. Age-adjusted cancer detection rate in "Universities and Technical Schools" were significantly higher than in "Factories", suggesting environment factor or diet might be associated with the prostate cancer. PMID- 15787335 TI - A case report: renal metastasis of prostate cancer. AB - In this report, a rare case of renal metastasis of prostate cancer in a 55-year old man is presented. PMID- 15787334 TI - Does asymptomatic inflammation increase PSA? A histopathological study comparing benign and malignant tissue biopsy specimens. AB - AIM: To determine the effect of inflammation on postage-specific antigen (PSA) levels in patients who were performed transrectal ultrasonography (TRUSG) guided prostate biopsy because of high PSA levels. MATERIALS AND METHOD: 376 tissue specimens of 47 patients with BPH and 176 tissue specimens of 22 patients with prostate adenocancer were analysed histopathologically. Digital rectal examination, TRUSG, and PSA measurements were performed to all the patients before biopsy. Sextant biopsy was performed. Two pathologists examined all the biopsy specimens in blinded fashion. Inflammation pattern was categorized as glandular, periglandular, stromal and perivascular and intensity of inflammation graded from 1 to 3. RESULTS: The mean age of the patients was 66.2 years. The mean serum PSA level of the initial biopsy was 8.7 ng/ml in the BPH and 13.4 ng/ml in the prostate Ca. No relation was found between the serum total PSA levels and prostate volume or patient's age in BPH patients (P = 0.258). In BPH patients stromal, periglandular, glandular inflammations did not increase PSA levels. Perivascular inflammation increased PSA levels significantly. CONCLUSION: The inflammation in perivascular field increases serum PSA levels (P = 0.007). Although high serum PSA levels shows correlation with cancer in biopsy, there is no such significant relation with rebiopsy results and PSA levels at last studies. Because of this, these patients' biopsy samples must be reinspected for infection findings and atipic biopsy or high grade PIN. The result of our study showed that histological perivascular infection within the prostatic gland is a significant factor to serum PSA levels in BPH. If you have negative biopsy sample, pathology must indicate the place of the infection. Follow up of this patient and rebiopsy time are decided according to the result of this negative biopsy sample. PMID- 15787336 TI - An HIV-negative penil Kaposi's sarcoma. AB - Kaposi's sarcoma is a tumour of reticuloendothelial system. Non-epidemic Kaposi's sarcoma limited to the penile involvement should be aggressively treated because it is rarely associated with diffuse organ involvement localized surgical excision or small-field external beam or electron beam radiation has been effective. In this case, we presented a man 53 years old, who presents with a papular indolent lesion on the glans penis, which is reported as a Kaposi sarcoma after excision of the lesion. PMID- 15787337 TI - A huge unusual mass on the penile skin: acrochordon. AB - Acrochordons are flesh-coloured pedunculated lesions which occur in areas of skin folds. Although they are common in other sites of the body, we report the first case of huge penile acrochordon in the literature. Clinical, pathological and surgical findings of this lesion were presented and discussed. PMID- 15787338 TI - The use of isolated sigmoid colon segment for vaginal replacement in young adults. AB - PURPOSE: Vaginoplasty for congenital atresia, a component of the Mayer-Rokitansky Kuster syndrome, or for gender confirmation, may be achieved by several techniques. This report focuses on the efficacy of rectosigmoid neocolporrhaphy (RSNC) for primary vaginal replacement. MATERIALS AND METHODS: From 1990 to 2002 we evaluated six patients 16-33 years old (average age 23) who required vaginal replacement. The diagnoses included the Mayer-Rokitansky syndrome in two cases, classic bladder exstrophy in 1 and male pseudohermaphrodites in three cases. The vagina was reconstructed using a 15-cm isolated sigmoid segment placed between the bladder and rectum and anastomosed to the introitus in four patients. The Frank procedure was used in two patients. RESULTS: A minimum of 1 year of followup is available in five of the six patients (mean of 32 month). Patients treated with sigmoid vaginoplasty had functional neovagina. with excellent cosmetic results and without excessive mucous production or the need for routine dilation. Stenosis at the mucocutaneous junction in one patient with a sigmoid vagina was treated with Y-V plasty. In the two patients managed by nonoperative perineal self-dilatation, there was little success noted and the two patients became surgical candidates. CONCLUSIONS: Experience with this group of patients leads us to believe that isolated sigmoid segments provides a cosmetic, self lubricating neovagina with low rates of failure and revision, and without the need for routine dilation. PMID- 15787339 TI - Histopathological evaluation of the preputium in preschool and primary school boys. AB - OBJECTIVE: To evaluate the dermatopathology of the preputium in preschool and primary school children. MATERIAL AND METHODS: The study group consisted of 32 boys subjected to circumcision procedure for religious belief consecutively between June 2003 and September 2003 in our clinic. The removed foreskin is examined by the same pathologist in all cases and the presence of any dermatopathology was noted. RESULTS: The mean age of the children was 6+/-2 (4 12) years. Three (9.3%) boys had complete phimosis while 2 (6.2%) had partial phimosis. Among 27 (84.3%) cases without phimosis, the histopathological examination of the preputium revealed focal lichenoid cellular infiltrate with epidermal atrophy in 1 (3.1%) patient. This pattern was considered as early lichen sclerosus et atrophicus (LSA). Minor changes were reported in 1 (3.1%) boy namely minimal inflammation and oedema while 1 (3.1%) patient had more marked oedema and acute inflammatory cell infiltration (diagnosed as balanoposthitis). Three (9.3%) preputium specimens revealed evidence of chronic inflammation. Finally 2 (6.2%) boys showed increased melanin pigmentation in basal segment of the foreskin. In 5 (15.6%) patients with phimosis 1 (20%) (complete phimosis) had chronic inflammation and oedema while the other (20%) (partial phimosis) had chronic inflammatory infiltrate only. The rest of the boys with phimosis had normal preputial pathology. As a total, 22 (68.8%) of the 32 cases had totally normal preputial foreskin. CONCLUSION: The presence of foreskin in preschool and primary school children might rarely be associated with important inflammatory dermatoses like LSA even in the absence of phimosis. PMID- 15787340 TI - Paratesticular pleomorphic rhabdomyosarcoma in an adult. AB - Rhabdomyosarcoma (RMS) is the commonest variety of soft tissue sarcoma in children and young adults. It represents 6.5% of all malignant tumours in paediatrics with an annual incidence of 4 to 7 cases/million children. The primary paratesticular site is considered to have a good prognosis in comparison with other rhabdomyosarcomas, despite the frequency of retroperitoneal lymph node invasion. This superficial site allows rapid diagnosis and consequently often complete resection of the tumor. The multi-disciplinary treatment of paratesticular RMS has improved control of the disease with a 2-year survival of 80%. We herein report a case with paratesticular pleomorphic RMS, a rare and prognostically unfavorable variant of RMS, who is disease free for 60 months after management with multidisciplinary approach. PMID- 15787341 TI - Premature ejaculation rates and treatment needs in males aged 20 years and over attending urology outpatient clinic. AB - AIM: The purpose of this study was the assessment of the rates of premature ejaculation and desires for treatment in male patients 20 years and over attending urology outpatient clinic. METHODS: All subjects included in the study were asked whether they had a complaint of premature ejaculation; if so, whether they had sought a doctor's assistance; if not, why they had not; why they did not mention their complaint during the first visit; whether they desired treatment; and if not, why they did not. RESULTS: A total of 1608 subjects were questioned. Premature ejaculation was present in 333 (20.7%). Premature ejaculation was the primary complaint of 33 of those attending the urology outpatient clinic. The most common reasons for not seeking a doctor's assistance were failure to see the condition as a problem, and embarrassment. Fifty four (16.2%) of these patients did not desire treatment. The most common reason for not desiring treatment was regarding it as unnecessary at present, and old age. CONCLUSION: The questioning of all men aged 20 and over attending urology outpatient clinic concerning premature ejaculation will greatly increase diagnoses and treatments. PMID- 15787342 TI - Interdialytic weight gain and pulmonary membrane diffusing capacity in patients on hemodialysis. AB - BACKGROUND: Measurement of pulmonary diffusion capacity for carbon monoxide (DLCO) may be useful for assessing disease affecting the alveolar-capillary bed or the pulmonary vasculature. It was reported that hemodialysis (HD) therapy causes DLCO reduction via decrease of pulmonary capillary blood volume components. The aim of the study was to evaluate the effect of interdialytic weight gain on pulmonary function and especially DLCO. We further determined whether intravascular volume status, assessed by inferior vena cava diameter (IVCD) contributes to DLCO in patients on HD. METHODS: Routine pulmonary function testing including forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), FEV1/FVC, forced mid-expiratory flow rate (FEF25-75), DLCO IVCD index and other echocardiographic parameters were evaluated in 20 patients (mean age 48.6+/-18.3 years, mean dialysis duration 17.4+/-19.2 months) on chronic HD, 1 hour after HD and after an interdialytic period (1 hour before HD therapy). Single-breath DLCO measurements were corrected for hemoglobin concentration (cDLCO). RESULTS: Routine pulmonary function tests (spirometry) showed no significant changes in FEV1, FVC and FEF25-75 whereas a statistically significant fall in FEV/FVC was found. At the end of the interdialytic period a statistically significant increase in weight, IVCD index, left ventriculer diastolic diameter (LVDD), and diastolic blood pressure (DBP) were observed (P < 0.05). Using the single-breath DLCO, we found unchanged cDLCO at the end of the interdialytic period. There was no correlation of cDLCO with increases in weight, DBP, IVCD index, LVDD (P > 0.05). CONCLUSION: The accumulation of body water between dialyses has no significant influence on DLCO. PMID- 15787343 TI - Absence or decreased endogenous thiosulfaturia: a cause of recurrent calcium nephrolithiasis. AB - In two earlier publications we reported excellent therapeutic results in patients with recurrent calcium nephrolithiasis, using oral or intravenous i.v. sodium thiosulfate on the one hand, and on treatment of tumorus-soft periarticular tissues calcifications on the other. Thus, we considered useful to measure endogenous thiosulfaturia, using a specific HPLC method, in 25 healthy adult males and 25 patients with recurrent calcium nephrolithiasis. Healthy adult males excreted between 11 and 16 microM/24 hour of endogenous thiosulfate, while patients with recurrent calcium nephrolithiasis excreted significantly lower amounts of 6 and 10 microM/24 hour, except one patient, who did not excrete endogenous thiosulfate, reflecting probably a genetic abnormality. Thiosulfate is a unique agent for treatment of recurrent calcium nephrolithiasis as well as some other abnormal calcium depositions. Two doses of 5 mM sodium thiosulfate daily are therapeutically sufficient. PMID- 15787345 TI - Hypercoagulability risk factors in children with minimal change disease and the protective role of protein-C activity. AB - It is believed that thrombotic activity in nephrotic syndrome is due to an imbalance between procoagulant/thrombotic and anticoagulant/antithrombotic factors in plasma. The aim of this study was to investigate the hypercoagulability risk in childhood minimal change disease and to find possible protective mechanisms with respect to hemostasis. Twenty-six children with minimal change disease were enrolled in this study. All patients were evaluated during an attack and on remission. The control group consisted of 33 healthy children. During the attack period, prothrombosis parameters, total lipid, cholesterol, fibrinogen levels and platelet count increased significantly compared to levels in the remission period. This denotes that hyperviscosity increases thrombosis tendency. In the attack period, the significant increase of prothrombin fragments 1 + 2 which shows thrombin formation and thrombin antithrombin complex which causes prothrombin activation, are an indication of increased thrombosis risk. Five patients with lupus anticoagulant present and 7 patients with, activated protein-C resistance ratios carried an increased thrombosis risk. D-dimer level of fibrinolytic factors significantly increased during the attack period. These findings emphasize the existence of thrombotic activity causing the activation of the fibrinolytic system. The significant increase in protein-C activity in these patients represents one of the protective mechanisms against thrombosis. The decrease in tissue plasminogen activator and antiplasmin indicates the protective role of fibrinolytic activity. Consequently, an increase in the protein-C activity is one of the protective mechanisms. The fibrinolytic system also plays an important role in preventing thrombotic activity in these patients. PMID- 15787346 TI - Chronic peritoneal dialysis in the tenth decade of life. AB - In this retrospective study we present our experience with chronic peritoneal dialysis in nine patients with ESRD in their 10th decade of life (> or =90 years) at the Toronto Western Hospital. A family member or a private nurse assisted all patients in dialysis procedure. The co-morbid illnesses, survival, hospitalizations and complications related or unrelated to peritoneal dialysis were reviewed. Four patients started dialysis before and five after their 90th birthday, their mean age was 90.61+/-4.04 years. All patients had three or more co-morbid illnesses at the start of dialysis. Total duration of PD treatment was 210 patient months with a median duration of 25 months (range 4-68 months). Of the nine patients, four died after a mean follow up of 38.5 months on dialysis. Of the remaining five, one was transferred to hemodialysis after remaining for 10 months on peritoneal dialysis and the other four are continuing on PD for a mean duration of 9.25 months. Peritonitis (1/13.4 patient months) and exit site infection (1/100.5 patient months) responded to treatment. Hospitalization rate was one admission per 2.5 patient years. Most often, the cause of hospitalization was unrelated to PD, e.g., cardiovascular events, pneumonia and peripheral vascular disease etc. Patient survival at 1, 3 and 5 years was 88%, 58% and 24% respectively. The technique survival was 69%, 47% and 23% at 1, 3 and 5 years respectively. We conclude that continuous peritoneal dialysis is a safe and suitable treatment even in nonagenarians (> or =90 years) ESRD patients. PMID- 15787344 TI - Oral supplement of six selective amino acids arrest progression renal failure in uremic patients. AB - Certain amino acids such as glycine, L-aspartic acid, L-glutamic acid, L glutamine, L-histidine and L-arginine taken orally by normal adults or patients with renal failure increase glomerular filtration rate (GFR). Twelve nondiabetic patients suffering from glomerulonephritis confirmed by renal biopsy previously, with creatinine clearances ranging from 15 to 24 ml minute/1.73, and on low protein diet 0.6 g/ kg/day, received an amino acid supplement daily in 2 or 3 doses for 1 year. At 4, 8 and 12 months creatinine clearance increased slightly (NS, NS, NS), 24 hour urine volume increased (P < or = 0.001, 001, 0.001), 24 hour albuminuria decreased (P < 0.001, 0.001, 0.001), serum urea increased (NS, NS, NS) serum albumin increased (NS, 0.05, 0.05), total cholesterol decreased slightly (NS, NS, 0.01), HDL increased slightly (0.05, 0.05, 0.05), LDL decreased (NS, 0.001, 0.001) triglycerides decreased (0.001, 0.001, 0.001), Apo B remained unchanged (NS, NS, NS), ROS/H2O2 decreased (0.001, 0,001, 0.001), Hct increased (NS, 0.01, 0.01) Hb increased (0.05, 0.05, 0.05), and serum phosphate decreased (0.01, 0.01, 0.01). After removal of supplements at the end of the year all parameters remained unchanged. We believe that a large controlled study should be undertaken to confirm these most encouraging findings. PMID- 15787347 TI - Characteristics of end stage renal disease diabetic patients in two countries with different socioeconomic conditions. AB - PURPOSE: To identify differences that may affect morbidity and mortality of type 2 diabetic patients reaching ESRD between countries with different socioeconomic conditions. METHODS: Comparison of clinical and laboratory features between 21 Nigerian (N) and 57 American patients (A) reaching ESRD over a 30 month period. RESULTS: Differences were noted in age at ESRD (N, 55.5+/-9.8; A, 64.5+/-9.6 years), duration of diabetes (N, 5.2+/-2.8, A: 14.9+/-4.9 years), body mass index (N, 24.5+/-4.1; A; 27.6+/-6.3 kg/m2), prevalence of left ventricular hypertrophy (N; 14%; A, 89%) and ischemic heart disease (N, 26%; A, 67%), blood pressure (N, [166.2+/-26.7]/[98.6+/-16.5] mmHg; A, [146.8+/-23.6]/[72.5+/-13.3] mmHg), creatinine clearance (N, 6.1+/-3.6; A, 14.8+/-3.5 ml/min), urine protein excretion (N, 1.2+/-0.7; A, 6.1+/-4.9 g/24-h), hematocrit (N, 28.0+/-6.0; A, 35.0+/-5.0%), serum glucose (N, 5.6+/-1.6; A, 10.5+/-5.5 mmol/l), and serum cholesterol (N, 5.32+/-2.57; A, 4.19+/-1.16 mmol/l) (all at P < or = 0.05). Differences were also found in the number of antihypertensive medications (N 1.4+/-0.6; A 2.4+/-1.2 per patient), and use of medications for diabetes (N 29%, A 79%), statins (N zero, A 61 %) and erythropoietin (N zero, A 39%). 72% of the A, but none of the N patients had a functional dialysis access prior to ESRD. CONCLUSIONS: Between A and N patients reaching ESRD, there are differences in clinical features and laboratory values that may affect morbidity, mortality and impact on the health care resources. These differences indicate areas where further studies that could assist in the planning for ESRD care in both Nigeria and USA are required. PMID- 15787349 TI - Ibandronate improves hyperphosphatemia in dialysis patients with hyperparathyroidism. PMID- 15787348 TI - Menopausal hormone therapy: is there evidence for cardiac protection?. AB - Menopausal hormone therapy currently has no evidence-based role in the primary or secondary prevention of cardiovascular disease in women. Repeat analysis of the observational data that corrected for socioeconomic status, educational level, and coronary risk factors; as well as information from randomized clinical trials of menopausal hormone therapy both for healthy women and for women with established coronary heart disease and stroke support this statement. This approach is corroborated by recommendations from the US Food and Drug Administration and the recently released Guidelines for Cardiovascular Prevention for Women of the American Heart Association. The latter Guidelines display the lifestyle and pharmacologic interventions that are appropriate for cardiovascular disease risk reduction for women. PMID- 15787350 TI - Nanoscale environmental science and technology: challenges and opportunities. PMID- 15787351 TI - Perchlorate report doesn't dispel controversy. PMID- 15787352 TI - China's wild card on transgenic tree front. PMID- 15787353 TI - Transgenic trees: opening Pandora's box? PMID- 15787354 TI - A fresh look at curbing greenhouse gases. PMID- 15787355 TI - A greener route to gold nanoparticles. PMID- 15787356 TI - Environmentally responsible development of nanotechnology. PMID- 15787357 TI - Preparation of a novel TiO2-based p-n junction nanotube photocatalyst. AB - TiO2 nanotube semiconductors contain free spaces in their interior that can be filled with active materials such as chemical compounds, enzymes, and noble metals, giving them a fundamental advantage over colloids. Although the unique shape of semiconductor nanotubes makes them promising for a range of potential applications, significant developmental research is required. In this research, a novel TiO2 nanotube photocatalyst was prepared that has a p-n junction. The photocatalyst particle surface is physically divided into reduction and oxidation surfaces, which poses a potential driving force for the transport of photogenerated charge carriers. The structure of this nanotube catalyst was characterized using a scanning electron microscope (SEM) and X-ray diffraction (XRD). The catalyst activity was evaluated by coating the catalyst on HEPA filters and determining the destruction rate of toluene in air. The p-n junction nanotube catalyst was shown to have a much higher photocatalytic destruction rate than that of commercially available, nonnanotube structured material, and a higher destruction rate for nanotube catalysts that did not contain a p-n junction. PMID- 15787358 TI - Sensitive detection of sulfhydryl groups in surface-confined metallothioneins and related species via ferrocene-capped gold nanoparticle/streptavidin conjugates. AB - Metallothionein (MT), a cysteine-rich metalloprotein that is purported to play an important role in heavy metal accumulation and detoxification, and its related peptidic species were attached onto dithiobissuccinimidyl propionate self assembled monolayers. The spatially accessible sulfhydryl groups present in these immobilized biomolecules, tagged with N-biotinoyl-N'-[6 maleimidohexanoyl]hydrazide, were detected voltammetrically at a sensitive level via the use of ferrocene (Fc)-capped gold nanoparticle/streptavidin conjugates. The method was established first by examining relatively simple peptides (e.g., glutathione). For the hexapeptidic species that resembles the N-terminus of MT with a sequence of Lys-Cys-Thr-Cys-Cys-Ala, concentration levels as low as 0.050 nM can be determined. Such a remarkable sensitivity is attributed to the presence of a large number of Fc caps present at each gold nanoparticle, which enhances the detection of a small number of surface-bound sulfhydryl groups. Microgravimetric measurements, performed with a quartz crystal microbalance, were used in tandem with voltammetry to quantify the number of tagged sulfhydryl groups. Through extraction of the metals present in MT adsorbate, it is demonstrated that this amplified voltammetric detection is also suitable for the investigation of the variation of the number of sulfhydryl groups present at an electrode and sensitive to the change of surface structure of an immobilized biomolecule. This work represents a new method for the determination of sulfhydryl groups inherent in surface-bound proteins or peptides and can facilitate the study on the environmental issues related to MTs. PMID- 15787359 TI - Development of improved materials for environmental applications: nanocrystalline NaY zeolites. AB - Two nanocrystalline NaY samples were synthesized with Si/Al ratios of 1.8 and crystal sizes of 23 and 50 nm, respectively. The synthesized NaY zeolites were characterized by powder X-ray diffraction, scanning electron microscopy, nitrogen adsorption isotherms, silicon solid-state magic angle spinning NMR and FTIR spectroscopy. A commercial NaY sample was analogously characterized for comparison with the synthesized nanocrystalline NaY. FTIR spectroscopy of adsorbed pyridine was used to elucidate the adsorption sites on the different NaY samples. More Bronsted acid sites and more silanol sites were detected on the nanocrystalline NaY zeolites, relative to the commercial NaY. The nanocrystalline NaY exhibited increased adsorption capacities for representative pollutant molecules, such as toluene (approximately 10%) and nitrogen dioxide (approximately 30%), relative to commercial NaY. Functionalization of nanocrystalline NaY was examined as a method for tailoring the properties of nanocrystalline zeolites for specific environmental applications through the control of zeolite properties, such as hydrophobicity. PMID- 15787360 TI - Characterization and properties of metallic iron nanoparticles: spectroscopy, electrochemistry, and kinetics. AB - There are reports that nano-sized zero-valent iron (Fe0) exhibits greater reactivity than micro-sized particles of Fe0, and it has been suggested that the higher reactivity of nano-Fe0 may impart advantages for groundwater remediation or other environmental applications. However, most of these reports are preliminary in that they leave a hostof potentiallysignificant(and often challenging) material or process variables either uncontrolled or unresolved. In an effort to better understand the reactivity of nano-Fe0, we have used a variety of complementary techniques to characterize two widely studied nano-Fe0 preparations: one synthesized by reduction of goethite with heat and H2 (Fe(H2)) and the other by reductive precipitation with borohydride (Fe(BH)). Fe(H2) is a two-phase material consisting of 40 nm alpha-Fe0 (made up of crystals approximately the size of the particles) and Fe3O4 particles of similar size or larger containing reduced sulfur; whereas Fe(BH) is mostly 20-80 nm metallic Fe particles (aggregates of <1.5 nm grains) with an oxide shell/coating that is high in oxidized boron. The FeBH particles further aggregate into chains. Both materials exhibit corrosion potentials that are more negative than nano-sized Fe2O3, Fe3O4, micro-sized Fe0, or a solid Fe0 disk, which is consistent with their rapid reduction of oxygen, benzoquinone, and carbon tetrachloride. Benzoquinone-which presumably probes inner-sphere surface reactions-reacts more rapidly with FeBH than Fe(H2), whereas carbon tetrachloride reacts at similar rates with FeBH and Fe(H2), presumably by outer-sphere electron transfer. Both types of nano-Fe0 react more rapidlythan micro-sized Fe0 based on mass-normalized rate constants, but surface area-normalized rate constants do not show a significant nano-size effect. The distribution of products from reduction of carbon tetrachloride is more favorable with Fe(H2), which produces less chloroform than reaction with Fe(BH). PMID- 15787361 TI - High resolution electron microscopy structural studies of organo-clay nanocomposites. AB - Engineering of clay nanocomposite materials by modification of their surfaces can enable the control of retention, transport, and persistence of toxic chemicals in the geosystem. The properties and interactions of clay nanocomposites have been widely studied, but little information exists on their microstructure at a range of scale extending down to atomic dimensions. The pairing of Na-montmorillonite clay with organic cations as well as with the herbicide fluridone, chosen as a model for an organic pollutant, was studied. Three organic cations were selected: hexadecyltrimethylammonium, benzyltrimethylammonium, and benzyltriethylammonium at 0%, 60%, and 100% of cation exchange capacity (CEC) loadings. A detailed microstructural analysis of the organo-clay nanocomposites and of the fluridone nanocomposites was undertaken by high-resolution transmission electron microscopy (HRTEM) and X-ray energy-dispersive spectroscopy (EDS). Morphological observations and chemical analyses were performed simultaneously on the same sample. The combined HRTEM and EDS measurements strongly suggest (a) heterogeneous local intercalation of the organic cations manifested by a range in the measured d001 spacing, implying random expansion of the clay layered structure with increased loading of the organic cations; (b) intercalation within the external layers, which is thoroughly influenced by local defect microstructure and/or edge availability of the montmorillonite nanoparticles as well as by the molecular structure of the intercalating organic cation. Additional intercalation of fluridone molecules did not affect the structure (d001 spacing) of the organo-clay nanocomposites. PMID- 15787362 TI - Heteroepitaxial nucleation and oriented growth of manganese oxide islands on carbonate minerals under aqueous conditions. AB - Manganese redox cycling and the accompanying dissolution and precipitation reactions are important processes in natural waters. In the present study, Mn2+ (aq) is reacted with O2(aq) at circumneutral pH to form manganese oxide islands on the (1014) surface of MnCO3. The islands grow heteroepitaxially. The effects of the substrate surface morphology, the substrate atomic structure, and the aqueous concentration of Mn2+ are investigated. On terraces, rhombohedral oxide islands form with 90 degrees rotation relative to the crystallographic axis of the underlying carbonate substrate. Although the island heights self-limit between 2 and 3 nm depending on reaction conditions, the islands grow laterally to several square microns before separate islands collide and coalesce. The islands do not grow over substrate steps or down dissolution-pit edges. Comparison studies done with MgCO3 and CaCO3 show that the former also promotes heteroepitaxial growth whereas the latter does not. This difference is explained by the relative bond length mismatch between the structures of the carbonate substrates and the atomic structures of manganese oxides. A free energy model is also presented to explain why the heights of the manganese oxide islands self limit. Our results provide an improved basis both for the development of predictive models of contaminant fate and transport and for the modeling of hydraulic flow through carbonate aquifers. PMID- 15787363 TI - Using a surface complexation model to predict the nature and stability of nanoparticles. AB - Nanoparticles are discrete nanometer-scale assemblies of atoms and have dimensions between those characteristic of ions and those of macroscopic materials. These minerals commonly possess extremely large specific surface areas and surface adsorption capacities for foreign ions. Due to the large specific surface area and large fraction of surface atoms, the natures of nanoparticles are expected to be modified by the adsorption (surface complexation) process. In this paper,we discuss theoretically the stability of nanoparticles that make the surface complex with foreign ions. The principal theoretical assumption is that the surface complexation occurs at the bulk of the nanoparticles, as in a solid solution. The surface complexation affects two aspects of the intrinsic stability of the nanoparticles simultaneously: one is the composition of the nanoparticles; the other is the free energy of formation of nanoparticles. The solubility of hydrous ferric oxide (HFO) was estimated by using surface complexation modeling coupled with published data of the free energy of formation of the relevant components. The solubility modeling of surface-charged (H+ or OH- sorbed) HFO mechanistically and quantitatively explained the observed nonintegral behavior of the solubility of HFO. Moreover, solubility modeling of anion (SO4(2-), PO4(3-), and As(V)) sorption by HFO showed that the sorption process strongly influences the stability of the nanoparticles. This result implies that geochemical modeling leads to the erroneous prediction of a natural system if the effect of the sorption process is not taken into account. PMID- 15787364 TI - Detection of heavy metal ions in drinking water using a high-resolution differential surface plasmon resonance sensor. AB - We have built a high-resolution differential surface plasmon resonance (SPR) sensor for heavy metal ion detection. The sensor surface is divided into a reference and sensing areas, and the difference in the SPR angles from the two areas is detected with a quadrant cell photodetector as a differential signal. In the presence of metal ions, the differential signal changes due to specific binding of the metal ions onto the sensing area coated with properly selected peptides, which provides an accurate real-time measurement and quantification of the metal ions. Selective detection of Cu2+ and Ni2+ in the ppt-ppb range was achieved by coating the sensing surface with peptides NH2-Gly-Gly-His-COOH and NH2-(His)6-COOH. Cu2+ in drinking water was tested using this sensor. PMID- 15787365 TI - Quantification of the oxidizing capacity of nanoparticulate zero-valent iron. AB - Addition of nanoparticulate zero-valent iron (nZVI) to oxygen-containing water results in oxidation of organic compounds. To assess the potential application of nZVI for oxidative transformation of organic contaminants, the conversion of benzoic acid (BA) to p-hydroxybenzoic acid (p-HBA) was used as a probe reaction. When nZVI was added to BA-containing water, an initial pulse of p-HBA was detected during the first 30 min, followed by the slow generation of additional p HBA over periods of at least 24 h. The yield of p-HBA increased with increasing BA concentration, presumably due to the increasing 'ability of BA to compete with alternate oxidant sinks, such as ferrous iron. At pH 3, maximum yields of p-HBA during the initial phase of the reaction of up to 25% were observed. The initial rate of nZVI-mediated oxidation of BA exhibited a marked reduction at pH values above 3. Despite the decrease in oxidant production rate, p-HBA was observed during the initial reaction phase at pH values up to 8. Competition experiments with probe compounds expected to exhibit different affinities for the nZVI surface (phenol, aniline, o-hydroxybenzoic acid, and synthetic humic acids) indicated relative rates of reaction that were similar to those observed in competition experiments in which hydroxyl radicals were generated in solution. Examination of the oxidizing capacity of a range of Fe0 particles reveals a capacity in all cases to induce oxidative transformation of benzoic acid, but the high surface areas that can be achieved with nanosized particles renders such particles particularly effective oxidants. PMID- 15787366 TI - Adsorption enhancement mechanisms of silica-titania nanocomposites for elemental mercury vapor removal. AB - A novel nanocomposite that combines high-surface area silica with the photocatalytic properties of titania has been developed that allows for effective capture of elemental mercury vapor. The adsorption capability of the developed material has been found to improve after periods of photocatalytic oxidation. In this study, the mechanisms for adsorption enhancement were identified. BET nitrogen adsorption and mercury porosimetry were used to evaluate pore structure, and the results suggest that a decrease in contact angle was likely to be responsible for improved mercury capture over time. Contact angle measurements showed a significant change of more than 10 degrees, indicating greater attraction to mercury for the used pellets due to deposited mercuric oxide. ICP and TGA analyses showed that mercury was captured as both elemental mercury (Hg0) and mercuric oxide (HgO). In addition, it was shown that pellets used for nearly 500 h still showed greater than 90% removal efficiency and had an average capacity of 10 mg of Hg/g based on mass balance calculations, while some pellets had a capacity over 30 mg of Hg/g according to ICP and TGA analyses. Mercuric oxide doped pellets removed 100% of elemental mercury without pretreatment. The superior mercury removal efficiency combined with various advantages of the novel composite demonstrates its use as an effective alternative to conventional activated carbon injection technology. PMID- 15787367 TI - Linker-based bio-compatible microemulsions. AB - In this work we have studied the formulation of biocompatible microemulsions using lecithin as the main surfactant and bio-compatible linker molecules (hexyl polyglucoside asthe hydrophilic linker and sorbitan monoleate as the lipophilic linker). These bio-compatible systems are discussed as potential substitutes for chlorinated solvents in dry-cleaning applications and as solvent delivery systems for pharmaceutical applications. Formulation parameters and conditions were evaluated using isopropyl myristate (IPM) as the model oil. It was found that the proposed linker-based formulations were able to form alcohol-free microemulsions while achieving higher solubilization capacity than similar systems reported in the literature. In addition, these lecithin/linker formulations were able to form microemulsions with a wide range of oils, from polar chlorinated hydrocarbons to hydrophobic oils such as squalene. These microemulsions were achieved under isotonic conditions (0.9% NaCl) by only varying the relative proportions of the linkers. The "solvency" power of these bio-compatible formulations was tested for the removal of hexadecane (used as model oil) from cotton fabrics and compared to the solvency power of a typical dry cleaning solvent tetrachloroethylene (PCE). While PCE and the linker-based lecithin formulation removed the same amount of hexadecane at low loading ratios (less than 1% oil volume fraction), at higher loading ratios the linker-based lecithin formulation retained its oil removal capacity while the efficiency of the PCE system declined rapidly. These initial results thus demonstrate the remarkable oil solubilization capacity of these bio compatible linker-based lecithin formulations and illustrate their potential as environmentally friendly replacements for organic solvents. PMID- 15787368 TI - Use of nanosized catalysts for transformation of chloro-organic pollutants. AB - A new method to transform anthropogenic, chloro-organic compounds (COC) by use of nanosized molecular catalysts immobilized in sol-gel matrixes is presented. COC represent a serious threat to soil and groundwater quality. Metalloporphyrinogens are nanometer sized molecules that are known to catalyze degradation of COC by reduction reactions. In the current study, metalloporphyrinogens were immobilized in sol--gel matrixes with pore throat diameters of nanometers. The catalytic activity of the matrix arrays for anaerobic reduction of tetrachloroethylene (PCE), trichloroethylene (TCE), and carbon tetrachloride (CT) was examined. Experiments were performed under conditions pertinent to groundwater systems, with titanium citrate and zero-valent iron as electron donors. All chloroorganic compounds were reduced in the presence of several sol-gel-metalloporphyrinogen hybrids (heterogeneous catalysts). For example, cobalt-5,10,15,20-(4 hydroxyphenyl)-21H,23H-porphine (TP(OH)P-Co) and cyanocobalamin (vitamin B12) reduced CT concentrations to less than 5% of their initial values in a matter of hours. Cyanocobalamin was found to reduce PCE to trace amounts in less than 48 h and TCE to less than 25% of its initial concentration in 144 h. The reactions were compared to their homogeneous (without sol-gel matrix) analogues. The reduction activity of COC for the homogeneous and heterogeneous systems ranged between similar reactivity in some cases to lower reduction rates for the heterogeneous system. These lower rates are, however, compensated by the ability to encapsulate and reuse the catalyst. Experiments with cyanocobalamin showed that the catalyst could be reused over at least 12 successive cycles of 24 h each. PMID- 15787369 TI - Removal of arsenic(III) from groundwater by nanoscale zero-valent iron. AB - Nanoscale zero-valent iron (NZVI) was synthesized and tested for the removal of As(III), which is a highly toxic, mobile, and predominant arsenic species in anoxic groundwater. We used SEM-EDX, AFM, and XRD to characterize particle size, surface morphology, and corrosion layers formed on pristine NZVI and As(III) treated NZVI. AFM results showed that particle size ranged from 1 to 120 nm. XRD and SEM results revealed that NZVI gradually converted to magnetite/maghemite corrosion products mixed with lepidocrocite over 60 d. Arsenic(III) adsorption kinetics were rapid and occurred on a scale of minutes following a pseudo-first order rate expression with observed reaction rate constants (K(obs)) of 0.07-1.3 min(-1) (at varied NZVI concentration). These values are about 1000x higher than K(obs) literature values for As(III) adsorption on micron size ZVI. Batch experiments were performed to determine the feasibility of NZVI as an adsorbent for As(III) treatment in groundwater as affected by initial As(III) concentration and pH (pH 3-12). The maximum As(III) adsorption capacity in batch experiments calculated by Freundlich adsorption isotherm was 3.5 mg of As(III)/g of NZVI. Laser light scattering (electrophoretic mobility measurement) confirmed NZVI As(III) inner-sphere surface complexation. The effects of competing anions showed HCO3-, H4SiO4(0), and H2P04(2-) are potential interferences in the As(III) adsorption reaction. Our results suggest that NZVI is a suitable candidate for both in-situ and ex-situ groundwater treatment due to its high reactivity. PMID- 15787370 TI - Axial flow cyclone for segregation and collection of ultrafine particles: theoretical and experimental study. AB - In this study, an axial flow cyclone was designed, fabricated, and evaluated at different conditions of air flow rates (Q0) and low-pressure environments (P), especially for the segregation and collection of ultrafine particles. An evaporation/condensation type of aerosol generation system consisting of tube furnace and mixing chamber was employed to produce test aerosols. The test aerosol was then classified by a differential mobility analyzer (DMA) and number concentration was measured by a condensation nuclei counter (CNC) and an electrometer upstream and downstream of the cyclone, respectively. The s-shaped curve of the collection efficiency in submicron particle size range was obtained to be similar to the traditional cyclone found in the literatures when the particles were largerthan 40 nm at Q0 = 1.07, 0.455 L(STP)/min, and P = 4.8-500 Torr. The curve was found to be fitted very well by a semiempirical equation described in this paper. For particles smaller than 40 nm, however, the collection efficiency was unusually increased as the particle diameter was decreased due to the fact that the diffusion deposition becomes the dominant collection mechanism in the low-pressure conditions. A model composed of centrifugal force and diffusion deposition is presented and used to fit the experimental data. The cyclone was demonstrated to separate and collect ultrafine particles effectively in the tested vacuum conditions. PMID- 15787371 TI - Field demonstration of DNAPL dehalogenation using emulsified zero-valent iron. AB - This paper describes the results of the first field-scale demonstration conducted to evaluate the performance of nanoscale emulsified zero-valent iron (EZVI) injected into the saturated zone to enhance in situ dehalogenation of dense, nonaqueous phase liquids (DNAPLs) containing trichloroethene (TCE). EZVI is an innovative and emerging remediation technology. EZVI is a surfactant-stabilized, biodegradable emulsion that forms emulsion droplets consisting of an oil-liquid membrane surrounding zero-valent iron (ZVI) particles in water. EZVI was injected over a five day period into eight wells in a demonstration test area within a larger DNAPL source area at NASA's Launch Complex 34 (LC34) using a pressure pulse injection method. Soil and groundwater samples were collected before and after treatment and analyzed for volatile organic compounds (VOCs) to evaluate the changes in VOC mass, concentration and mass flux. Significant reductions in TCE soil concentrations (>80%) were observed at four of the six soil sampling locations within 90 days of EZVI injection. Somewhat lower reductions were observed at the other two soil sampling locations where visual observations suggest that most of the EZVI migrated up above the target treatment depth. Significant reductions in TCE groundwater concentrations (57 to 100%) were observed at all depths targeted with EZVI. Groundwater samples from the treatment area also showed significant increases in the concentrations of cis-1,2 dichloroethene (cDCE), vinyl chloride (VC) and ethene. The decrease in concentrations of TCE in soil and groundwater samples following treatment with EZVI is believed to be due to abiotic degradation associated with the ZVI as well as biodegradation enhanced by the presence of the oil and surfactant in the EZVI emulsion. PMID- 15787372 TI - Nanoporous silica-supported nanometric palladium: synthesis, characterization, and catalytic deep oxidation of benzene. AB - In this present study, nanoporous silica SBA-15 supported palladium catalysts are prepared through two different methods. The catalysts are employed for catalytic deep oxidation reaction of benzene at a high gas hourly space velocity of 100,000 h(-1). It is found that the traditional aqueous impregnation method has some difficulties and disadvantages in obtaining highly dispersed palladium active phases. Whereas, when a grafting procedure is employed, palladium tends to be highly dispersed as nanoparticles due to the confinement of the nanosized pore channels of the support materials. The catalysts prepared via the grafting procedure catalyze the benzene oxidation far more effectively than those prepared via aqueous impregnation method, and complete conversion of benzene can be achieved below 190 degrees C over the most active catalyst. The nanoporous silica supported palladium catalysts are promising materials for the control of some types of VOCs such as benzene. PMID- 15787373 TI - Actinide sequestration using self-assembled monolayers on mesoporous supports. AB - Surfactant templated synthesis of mesoporous ceramics provides a versatile foundation upon which to create high efficiency environmental sorbents. These nanoporous ceramic oxides condense a huge amount of surface area into a very small volume. The ceramic oxide interface is receptive to surface functionalization through molecular self-assembly. The marriage of mesoporous ceramics with self-assembled monolayer chemistry creates a powerful new class of environmental sorbent materials called self-assembled monolayers on mesoporous supports (SAMMS). These SAMMS materials are highly efficient sorbents whose interfacial chemistry can be fine-tuned to selectively sequester a specific target species, such as heavy metals, tetrahedral oxometalate anions, and radionuclides. Details addressing the design, synthesis, and characterization of SAMMS materials specifically designed to sequester actinides, of central importance to the environmental cleanup necessary after 40 years of weapons-grade plutonium production, as well as evaluation of their binding affinities and kinetics are presented. PMID- 15787374 TI - Incorporation of hydroxypyridinone ligands into self-assembled monolayers on mesoporous supports for selective actinide sequestration. AB - In this study, three isomers of hydroxypyridinones (1,2-HOPO, 3,2-HOPO, and 3,4 HOPO) were attached to self-assembled monolayers on mesoporous silica (SAMMS). The HOPO-SAMMS materials have superior solid adsorbents properties: they do not suffer from solvent swelling; their rigid, open pore structure allows rapid sorption kinetics; their extremely high surface area enables the installation of high functional density; and being silica-based, they are compatible with vitrification into a final vitreous waste form. Kinetics, equilibrium, and selectivity of the adsorptions of actinide on the HOPO-SAMMS at various pH values and in the presence of other metal cations, anions, and competing ligands are reported. Rapid sequestration of U(VI), Np(V), and Pu(IV) was observed. Very little competition from transition metal cations and common species was observed. PMID- 15787375 TI - TCE dechlorination rates, pathways, and efficiency of nanoscale iron particles with different properties. AB - Nanoscale Fe0 particles are a promising technology for in situ remediation of trichloroethene (TCE) plumes and TCE-DNAPL source areas, butthe physical and chemical properties controlling their reactivity are not yet understood. Here, the TCE reaction rates, pathways, and efficiency of two nanoscale Fe0 particles are measured in batch reactors: particles synthesized from sodium borohydride reduction of ferrous iron (Fe/B) and commercially available particles (RNIP). Reactivity was determined under iron-limited (high [TCE]) and excess iron (low [TCE]) conditions and with and without added H2. Particle efficiency, defined as the fraction of the Fe0 in the particles that is used to dechlorinate TCE, was determined under iron-limited conditions. Both particles had a core/shell structure and similar specific surface areas (approximately 30 m2/g). Using excess iron, Fe/B transformed TCE into ethane (80%) and C3-C6 coupling products (20%). The measured surface area normalized pseudo-first-order rate constant for Fe/B (1.4 x 10(-)2 L.h(-1).m(-2) is approximately 4-fold higher than for RNIP (3.1 x 10-(3) L.h(-1).m(-2). All the Fe0 in Fe/B was accessible for TCE dechlorination, and 92 +/- 0.7% of the Fe0 was used to reduce TCE. For Fe/B, H2 evolved from reduction of water (H+) was subsequently used for TCE dechlorination, and adding H2 to the reactor increased both the dechlorination rate and the mass of TCE reduced, indicating that a catalytic pathway exists. RNIP yielded unsaturated products (acetylene and ethene). Nearly half (46%) of the Fe0 in RNIP was unavailable for TCE dechlorination over the course of the experiment and remained in the particles. Adding H2 did not change the reaction rate or efficiency of RNIP. Despite this, the mass of TCE dechlorinated per mass of Fe0 added was similar for both particles due to the less saturated products formed from RNIP. The oxide shell composition and the boron content are the most likely causes for the differences between the particle types. PMID- 15787376 TI - Designing Pd-on-Au bimetallic nanoparticle catalysts for trichloroethene hydrodechlorination. AB - Alumina-supported palladium (Pd) catalysts have previously been shown to hydrodechlorinate trichloroethene (TCE) and other chlorinated compounds in water, at room temperature, and in the presence of hydrogen. The feasibility of this catalytic technology to remediate groundwater of halogenated compounds can be improved by re-designing the Pd material in order to increase catalytic activity. We synthesized and characterized Pd supported on gold nanoparticles (Au NPs) of different Pd loadings. In all cases, we found that these catalysts were considerably more active than Pd NPs, alumina-supported Pd, ard Pd-black (62.0, 12.2, and 0.42 L x g(Pd)(-1) x min(-1), respectively). There is a synergistic effect of the Pd-on-Au bimetallic structure, with the material with the highest TCE hydrodechlorination activity (943 L x g(Pd)(-1) x min(-1)) comprised of Au NPs partially covered by Pd metal. The Pd-on-Au bimetallic catalyst structure provides a new synthesis approach in improving the catalytic properties of monometallic Pd materials. The resulting nanoparticle-based materials should be highly suitable as hydrodehalogenation and reduction catalysts for the remediation of various organic and inorganic groundwater contaminants. PMID- 15787377 TI - Engineered polymeric nanoparticles for bioremediation of hydrophobic contaminants. AB - Sorption of hydrophobic organic contaminants, such as polycyclic aromatic hydrocarbons (PAHs), to soil has been shown to limit their solubilization rate and mobility. In addition, sequestration of contaminants by sorption to soil and by partitioning in nonaqueous phase liquids (NAPLs) reduces their bioavailability. Polymeric nano-network particles have been demonstrated to increase the "effective" solubility of a representative hydrophobic organic contaminant, phenanthrene (PHEN) and to enhance the release of PHEN from contaminated aquifer material. In this study, we investigate the usefulness of nanoparticles made from a poly(ethylene) glycol modified urethane acrylate (PMUA) precursor chain, in enhancing the bioavailability of PHEN. PMUA nanoparticles are shown to increase the mineralization rate of PHEN crystal in water, PHEN sorbed on aquifer material, and PHEN dissolved in a model NAPL (hexadecane) in the presence of aquifer media. These results show that PMUA particles not only enhance the release of sorbed and NAPL-sequestered PHEN but also increase its mineralization rate. The accessibility of contaminants in PMUA particles to bacteria also suggests that particle application may be an effective means to enhance the in-situ biodegradation rate in remediation through natural attenuation of contaminants. In pump-and-treat or soil washing remediation schemes, bioreactors could be used to recycle extracted nanoparticles. The properties of PMUA nanoparticles are shown to be stable in the presence of a heterogeneous active bacterial population, enabling them to be reused after PHEN bound to the particles has been degraded by bacteria. PMID- 15787378 TI - Fullerol-sensitized production of reactive oxygen species in aqueous solution. AB - The relative production rate of reactive oxygen in aqueous solution sensitized by fullerol (a polyhydroxylated, water-soluble form of the fullerene C60) was measured and compared to known reactive oxygen sensitizers using an oxygen consumption method. The solutions were irradiated by polychromatic visible and ultraviolet light. Reactive oxygen species were generated under both visible and ultraviolet light sources. The greatest rates of oxygen consumption were observed at acidic pH. We show for the first time evidence of both singlet oxygen and superoxide production by fullerol under both UV and polychromatic light sources. PMID- 15787379 TI - Dendrimer enhanced ultrafiltration. 1. Recovery of Cu(II) from aqueous solutions using PAMAM dendrimers with ethylene diamine core and terminal NH2 groups. AB - This article discusses the feasibility of using dendrimer enhanced ultrafiltration (DEUF)to recover Cu(II) from aqueous solutions. Building upon the results of fundamental investigations of Cu(II) binding to PAMAM dendrimers with ethylenediamine (EDA) core and terminal NH2 groups, we combine (i) dead-end ultrafiltration (UF) experiments with (ii) atomic force microscopy (AFM) characterization of membrane fouling to assess the feasibility of using DEUF to recover Cu(II) from aqueous solutions. On a mass basis, the Cu(II) binding capacities of the EDA core PAMAM dendrimers are much larger and more sensitive to solution pH than those of linear polymers with amine groups. The dendrimer-Cu(II) complexes can be efficiently separated from aqueous solutions by ultrafiltration. The metal ion laden dendrimers can be regenerated by decreasing the solution pH to 4.0; thus enabling the recovery of the bound Cu(II) ions and recycling of the dendrimers. The UF measurements and AFM characterization studies show that EDA core PAMAM dendrimers with terminal NH2 groups have very lowtendency to foul the commercially available regenerated cellulose (RC) membranes evaluated in this study. The overall results of these experiments suggest that DEUF is a promising process for recovering metal ions such as Cu(II) from aqueous solutions. PMID- 15787380 TI - Cytotoxicity of carbon nanomaterials: single-wall nanotube, multi-wall nanotube, and fullerene. AB - A cytotoxicity test protocol for single-wall nanotubes (SWNTs), multi-wall nanotubes (with diameters ranging from 10 to 20 nm, MWNT10), and fullerene (C60) was tested. Profound cytotoxicity of SWNTs was observed in alveolar macrophage (AM) after a 6-h exposure in vitro. The cytotoxicity increases by as high as approximately 35% when the dosage of SWNTs was increased by 11.30 microg/cm2. No significant toxicity was observed for C60 up to a dose of 226.00 microg/cm2. The cytotoxicity apparently follows a sequence order on a mass basis: SWNTs > MWNT10 > quartz > C60. SWNTs significantly impaired phagocytosis of AM at the low dose of 0.38 microg/cm2, whereas MWNT10 and C60 induced injury only at the high dose of 3.06 microg/cm2. The macrophages exposed to SWNTs or MWNT10 of 3.06 microg/cm2 showed characteristic features of necrosis and degeneration. A sign of apoptotic cell death likely existed. Carbon nanomaterials with different geometric structures exhibit quite different cytotoxicity and bioactivity in vitro, although they may not be accurately reflected in the comparative toxicity in vivo. PMID- 15787382 TI - [Depression--the "express disease" of our times?]. PMID- 15787381 TI - Life cycle benefits of using nanotechnology to stabilize platinum-group metal particles in automotive catalysts. AB - Due to advances in nanotechnology, the approach to catalytic design is transitioning from trial-and-error to planned design and control. Expected advances should enable the design and construction of catalysts to increase reaction speed, yield, and catalyst durability while also reducing active species loading levels. Nanofabrication techniques enabling precise control over the shape, size, and position of nanoscale platinum-group metal (PGM) particles in automotive catalysts should result in reduced PGM loading levels. These reductions would decrease energy consumption, improve environmental quality, and contribute to sustainable resource usage. We estimate the amount of PGM required to meet U.S. vehicle emissions standards through 2030 based on current catalysttechnology. We then estimate the range of PGM that could be saved from potential nanotechnology advances. Finally, we employ economic input-output and process-based life cycle assessment models to estimate the direct and life cycle benefits from reducing PGM mining and refining. PMID- 15787383 TI - [Postpartum depression is not merely "Baby Blues"]. PMID- 15787384 TI - [Help for depressive diseases in childhood and adolescence]. PMID- 15787385 TI - [Suitable for professional uniform clothing]. PMID- 15787386 TI - [Healthy living--and the working world of the clinic]. PMID- 15787387 TI - [Nursing process documentation in the ENP (European Nursing care Pathways)]. PMID- 15787389 TI - [Looking forward to the Nursing Rights' Day. Financing continuing education and job safety]. PMID- 15787388 TI - [Getting old and living in Zimbabwe/Africa]. PMID- 15787390 TI - [Time off for nursing education]. PMID- 15787391 TI - [A moving letter from Indonesia]. PMID- 15787393 TI - [Work conditions even more difficult for nurses]. PMID- 15787394 TI - [The drama at Pau causes a resurgence of distress in the psychiatric sector]. PMID- 15787395 TI - [Top launching of the second version of the accreditation manual]. PMID- 15787396 TI - [Delegation of acts: only for family caregivers]. PMID- 15787397 TI - [Tuberculosis, a very precarious Parisian disease]. PMID- 15787398 TI - [Campaign against homophobia, even in the hospital]. PMID- 15787400 TI - [2004, what legal evaluation for the health world?]. PMID- 15787401 TI - [New codification of nursing texts, practical consequences]. PMID- 15787402 TI - [Leg ulcers followed in an urban practice: performance of a hydrocellular dressing, non-adhesive Biatain Ulcer]. PMID- 15787403 TI - [Gastrostomy in adults and children]. PMID- 15787404 TI - [Writing, nursing care and research]. PMID- 15787405 TI - [Letter to a nurse]. PMID- 15787406 TI - [What quality of language in nursing records?]. PMID- 15787407 TI - [Clinical research and the nurse's right to information]. PMID- 15787408 TI - [The question of research at the end of studies at the Institute]. PMID- 15787409 TI - [Writing to formalize nursing knowledge]. PMID- 15787410 TI - [Transmitting knowledge, a challenge for the future]. PMID- 15787411 TI - [Bibliography elements]. PMID- 15787412 TI - [Cardiovascular system drugs. 9/12. Antilipemics--II. Fibrates and colestyramine]. PMID- 15787413 TI - [1/6 Traffic accidents, the key numbers]. PMID- 15787414 TI - [II--Relationship care. 10. Persons who present with post traumatic stress syndrome after a rape]. PMID- 15787416 TI - [Scarcity of nurses, the temporary have health]. PMID- 15787415 TI - [Employment, the embarrassment of choice at the end of scarcity]. PMID- 15787417 TI - Why the NHS money-go-round is running out of steam. PMID- 15787418 TI - On rewarding consultants. PMID- 15787419 TI - Dutch healthcare. Why choice went Dutch. AB - The Netherlands, like the UK, has selected increased patient choice as the driver for reform of an expensive and overstretched health service. As the piece's author, Paul Neuburg, writes: 'With choice as their weapon, consumers are being enlisted as food soldiers in the struggle to raise healthcare standards and contain costs.' PMID- 15787420 TI - Data briefing. Impact of choice on hospital viability. PMID- 15787421 TI - Tsunami relief. After the wave. AB - With the official toll from the Boxing Day tsunami now standing at 300,000, Alexis Nolan and Emma Forrest talk to health service managers who gave more than money to the relief effort--they travelled to the disaster area and gave their time and expertise. The impact on their professional and personal lives was considerable. As Mathi Chandrakumar, clinical director of Kent Health Protection Unit, puts it: 'My heart is there, I feel very sad. I will probably go back.' PMID- 15787422 TI - On clinical management. PMID- 15787423 TI - Workplace stress. Pressure point. PMID- 15787424 TI - Office politics. Why you didn't get promoted. PMID- 15787425 TI - Tsunami tales. 'The water just welled up'. PMID- 15787426 TI - Dietary intake of macronutrients, micronutrients, and other dietary constituents: United States 1988-94. AB - OBJECTIVES: This report presents national estimates of dietary intakes of macronutrients, micronutrients, and other dietary constituents for persons 2 months and older, by sociodemographic variables. METHODS: The third National Health and Nutrition Examination Survey (NHANES III) (1988-94), the source of these data, was designed to provide information on the health and nutritional status of the civilian noninstitutionalized U.S. population. The analytic sample for these analyses included 29,105 participants with complete and reliable dietary recalls. RESULTS: This report provides mean, median, and standard error of the mean dietary intake data for the U.S. population, 1988-94. Dietary intake estimates were derived from NHANES III 24-hour recall data. Foods reported during the survey were coded using the U.S. Department of Agriculture (USDA) Survey Nutrient Database (SNDB) and the University of Minnesota Nutrition Coordinating Center (NCC) data. Food composition values for macronutrients, vitamins, minerals, and other food components are based on SNDB and NCC data. CONCLUSIONS: Data on nutrient intakes are a crucial piece of information toward the determination of total dietary intake. Together with anthropometric and laboratory data, information on nutrient intake can be used to assess nutritional status and elucidate the relationship between nutrition and health. PMID- 15787427 TI - Acutangulosides A-F, monodesmosidic saponins from the bark of Barringtonia acutangula. AB - Nine triterpene saponins, acutangulosides A-F (2-7), and acutanguloside D-F methyl esters (5a-7a) and a single triterpene aglycone (1) were isolated from a water extract of the bark of Barringtonia acutangula. Their structures were assigned on the basis of spectroscopic data. PMID- 15787428 TI - Antiinsectan decaturin and oxalicine analogues from Penicillium thiersii. AB - Three new oxalicine and decaturin analogues (6-8) have been isolated from organic extracts of Penicillium thiersii, along with the known compounds oxalicines A and B. These compounds (4-8) are members of a group of unique natural products containing terpenoid and pyridine moieties. The structures of the new compounds were elucidated by analysis of NMR and MS data. Most of these metabolites exhibit potent antiinsectan activity against the fall armyworm (Spodoptera frugiperda). PMID- 15787429 TI - Terphenylquinone inhibitors of the src protein tyrosine kinase from Stilbella sp. AB - Three new secondary metabolites, 2,5-dihydroxy-3-(3,4-dihydroxyphenyl)-6-phenyl 1,4-benzoquinone (3), 2,5-dihydroxy-3-phenyl-6-(3,4,5-trihydroxyphenyl)-1,4 benzoquinone (4), and 2,7,8-trihydroxy-3-phenyl-1,4-dibenzofurandione (5), as well as two known natural products were isolated from the fungus Stilbella sp. strain 1586. The structures of these compounds were established by spectroscopic and chemical methods. The terphenylquinones 3-5 exhibited significant activity against human src protein tyrosine kinase. PMID- 15787430 TI - Dominicin, a cyclic octapeptide, and laughine, a bromopyrrole alkaloid, isolated from the Caribbean marine sponge Eurypon laughlini. AB - Dominicin, a new cyclic peptide, and the new bromopyrrole alkaloid laughine (4) have been isolated from the marine sponge Eurypon laughlini collected in Dominica. The structures of 1 and 4 were determined by a combination of spectroscopic analysis and chemical degradation. Single-crystal X-ray diffraction analysis confirmed the structure of dominicin (1). PMID- 15787431 TI - Cytotoxic sterol derivatives from a marine sponge Homaxinella sp. AB - A bioactivity-guided fractionation of a marine sponge Homaxinella sp. has led to the isolation of three new (1-3) highly degraded sterols and four new 6-O alkylated (6-9) sterols, along with known sterol derivatives. The degraded sterols (1-5) belong to the class incisterols, previously isolated from the marine sponge Dictyonella incisa. Mainly NMR and MS spectroscopic analyses established the gross structures of the new compounds. The relationship between the stereoisomerism of the side chain and HPLC retention time has also been discussed. The compounds were tested against a panel of five human solid tumor cell lines, and especially the degraded sterols (1-4) displayed significant cytotoxicity. PMID- 15787432 TI - Bioactive asterosaponins from the starfish Culcita novaeguineae. AB - Two new sulfated steroidal pentaglycosides (asterosaponins), novaeguinosides I (2) and II (3), along with the known regularoside B (1) were isolated from the starfish Culcita novaeguineae. Their structures were elucidated by extensive NMR techniques as well as chemical evidence. The new asterosaponins showed marginal in vitro cytotoxicity against two human tumor cell lines. PMID- 15787433 TI - Synthesis and biological evaluation of a natural ester sintenin and its synthetic analogues. AB - Synthesis of 3-(3,4-dimethoxyphenyl)propyl-3-(3,4-dimethoxyphenyl) propanoate (18), a cytotoxic natural ester, was carried out by a convenient synthetic path with a total yield of 49%. Sixteen of its analogues (19-34) were also prepared. Seventeen unsaturated derivatives of 18, compounds 1-17, were also synthesized to examine the structure-activity relationship of this type of ester. All of the synthetic compounds were passed through the cytotoxicity screenings on human tumor cell lines, such as PC-3, Hela, A549, BEL7404, CNE, and KB. Some of the esters exhibited moderate inhibitory effects on these tumor cell lines. The phenolic derivatives exhibited the highest cytotoxicity among these derivatives, while the unsaturated esters were more cytotoxic than the saturated analogues. Some of the compounds also exhibited inhibition on alpha-glucosidase. PMID- 15787434 TI - Chinikomycins A and B: isolation, structure elucidation, and biological activity of novel antibiotics from a marine Streptomyces sp. isolate M045. AB - In our screening of marine Streptomycetes for bioactive principles, two novel antitumor antibiotics designated as chinikomycins A (2a) and B (2b) were isolated together with manumycin A (1), and their structures were elucidated by a detailed interpretation of their spectra. Chinikomycins A (2a) and B (2b) are chlorine containing aromatized manumycin derivatives of the type 64-pABA-2 with an unusual para orientation of the side chains. They exhibited antitumor activity against different human cancer cell lines, but were inactive in antiviral, antimicrobial, and phytotoxicity tests. PMID- 15787435 TI - Bioactive benzophenones from Garcinia xanthochymus fruits. AB - A MeOH extract of Garcinia xanthochymus fruits was subjected to activity-guided fractionation, yielding two new benzophenones, guttiferone H (1) and gambogenone (2). Compound 1 contains a seven-membered ring attached to the bicyclo[3.3.1]nonane system at positions 7 and 8 and displayed cytotoxicity in the SW-480 colon cancer cell line (IC(50) = 12 microM). Compound 2 has a novel benzophenone bicyclo[3.3.2]decane system and displayed cytotoxicity in the SW-480 colon cancer cell line (IC(50) = 188 microM). Both 1 and 2 induced apoptosis in SW-480 colon cancer cells and displayed antioxidant activity in the 1,1-diphenyl 2-picrylhydrazyl (DPPH) assay (IC(50) = 64 and 38.7 microM, respectively). The structures of 1 and 2 were established by 1D and 2D NMR data analysis. Eleven known compounds, aristophenone A, alloathyriol, amentoflavone, 3,8' '-biapigenin, cycloxanthochymol, (+/-)-fukugetin, (+/-)-fukugiside, guttiferone E, isoxanthochymol, (+/-)-volkensiflavone, and xanthochymol, were also obtained. The 11 known compounds were also tested against SW-480 colon cancer cells and in the DPPH assay. PMID- 15787436 TI - Phenolic constituents of the rhizomes of the Thai medicinal plant Belamcanda chinensis with proliferative activity for two breast cancer cell lines. AB - From the rhizomes of Belamcanda chinensis, three new compounds, belalloside A (1), belalloside B (2), and belamphenone (3), along with 13 known compounds, resveratrol (4), iriflophenone (5), irisflorentin (6), tectorigenin (7), irilin D (8), tectoridin (9), iristectorin A (10), iristectorin B (11), hispiduloside, androsin, irigenin, iridin, and jaceoside, have been isolated and characterized. Isolates were evaluated for their cell proliferation stimulatory activity against the MCF-7 and T-47D human breast cancer cell lines. Along with 4, 5, 7, and 9, 3 was shown to stimulate not only MCF-7 but also T-47D human breast cancer cell proliferation. PMID- 15787438 TI - Cytotoxic 4-phenylcoumarins from the leaves of Marila pluricostata. AB - Bioassay-guided fractionation of the CH(2)Cl(2) extract of the leaves of Marila pluricostata led to the isolation of 17 naturally occurring 4-phenylcoumarins, three of them, 5-hydroxy-8,8-dimethyl-4-phenyl-9,10-dihydro-8H-pyrano-[2,3 f]chromen-2-one (1), 5-hydroxy-8,8-dimethyl-4-phenyl-6-propionyl-9,10-dihydro-8H pyrano-[2,3-f]chromen-2-one (2), and 5,7-dihydroxy-8-(3-methylbut-2-enyl)-4 phenylchromen-2-one (3), are new natural compounds; the remaining (4-17) are known mammea-type coumarins. Their structures were established by spectroscopic means. All compounds were tested in cytotoxicity assays against the MCF-7, H-460, and SF-268 human cancer cell lines. PMID- 15787437 TI - Sesquiterpene phenylpropanoids from Ferula fukanensis and their nitric oxide production inhibitory effects. AB - Five new sesquiterpene phenylpropanoid derivatives, fukanedone A (1), fukanedone B (2), fukanedone C (3), fukanedone D (4), and fukanedone E (5), and a novel phenyl-oxo-acetate ester, fukaneketoester A (6), were isolated from a 80% aqueous methanol extract of the roots of Ferula fukanensis. The structures were elucidated on the basis of spectroscorpic evidence, especially heteronuclear multiple-bond connectivity (HMBC) and high-resolution MS. The sesquiterpene phenylpropanoid derivatives inhibited nitric oxide (NO) production and inducible NO synthase (iNOS) gene expression by a murine macrophage-like cell line (RAW 264.7), which was activated by lipopolysaccharide (LPS) and recombinant mouse interferon-gamma (IFN-gamma). PMID- 15787439 TI - Psychopharmacological profile of the alkaloid psychollatine as a 5HT2A/C serotonin modulator. AB - Behavioral effects of psychollatine, a new glycoside indole monoterpene alkaloid isolated from Psychotria umbellata, was investigated in models of anxiety, depression, memory, tremor, and sedation related to 5-HT and/or GABA neurotransmission. The GABA antagonist picrotoxin and the 5-HT2 antagonist ritanserin were used to examine the role of GABA and 5-HT2 receptors in psychollatine-induced effects. In the light/dark and hole-board models of anxiety, diazepam (0.75 mg/kg) and psychollatine (7.5 and 15 mg/kg) showed anxiolytic-like effect at doses that do not increase sleeping time nor alter spontaneous locomotor activity. The anxiolytic effect of psychollatine was prevented by prior administration of ritanserin, but not of picrotoxin, indicating that 5-HT2 but not GABA receptors are implicated. In the forced swimming model of depression, psychollatine (3 and 7.5 mg/kg) effects were comparable to the antidepressants imipramine (15 mg/kg) and fluoxetine (20 mg/kg). Psychollatine suppressed oxotremorine-induced tremors in all doses. In the step-down learning paradigm, diazepam (0.85 mg/kg), MK-801 (0.15 mg/kg), and psychollatine 100 mg/kg impaired the acquisition of learning and memory consolidation, without interfering with retrieval. It is concluded that the effects of psychollatine at the central nervous system involve serotonergic 5HT2(A/C) receptors. PMID- 15787440 TI - Homoflavonoids from Ophioglossum petiolatum. AB - Six homoflavonoids, ophioglonin (1), ophioglonin 7-O-beta-D-glucopyranoside (2), ophioglonol (3), ophioglonol prenyl ether (4), ophioglonol 4'-O-beta-D glucopyranoside (5), and isoophioglonin 7-O-beta-D-glucopyranoside (6), together with five known compounds, quercetin, luteolin, kaempferol, 3,5,7,3',4' pentahydroxy-8-prenylflavone, and quercetin 3-O-methyl ether, were isolated from Ophioglossum petiolatum. Their structures were elucidated by analysis of spectroscopic data and chemical evidence. Compounds 1 and quercetin 3-O-methyl ether showed slight anti-HBV surface antigen activity at 25 microM. PMID- 15787441 TI - An MS/MS library on an ion-trap instrument for efficient dereplication of natural products. Different fragmentation patterns for [M + H]+ and [M + Na]+ ions. AB - The structural novelty of lead compounds is very important in the agrochemical and pharmaceutical industries, and as such, natural products can be an important source. Taking into account that the isolation of lead compounds is very time consuming, the efficient and safe identification of compounds in microorganism and plant extracts isolated previously is essential. A suitable procedure for this task based on an HPLC system interfaced with an electrospray (ESI) source and a Thermo Finnigan LCQ deca XP plus ion-trap mass spectrometer was developed, and an extensive MS/MS spectral library of characterized natural products was built up. This report summarizes the parameters used for acquiring the library spectra and discusses current limitations of the NIST library and search algorithm. The advantages of the newly introduced Mass Frontier 4.0 for the search of MS/MS product-ion spectra are discussed. Different mechanisms for fragmentation of some [M + H](+) and [M + Na](+) ions that were found are proposed. Oligomycin A, a macrolide antibiotic, exhibits different fragmentation mechanisms in positive and negative ion modes. The cleavage of the ester bond is the preferred mechanism in the positive ion mode, whereas two different pathways one showing a rare retro-Michael-addition-are observed in the negative ion mode. PMID- 15787442 TI - Dihydrochalcones from the leaves of Pieris japonica. AB - Six new dihydrochalcones, 3-hydroxyasebotin (5), asebogenin 2'-O-beta-D-ribohexo 3-ulopyranoside (6), 2' '-acetylasebotin (7), 3',4,5'-trihydroxy-4' methoxydihydrochalcone 3',5'-di-O-beta-D-glucopyranoside (8), and pierotins A (9) and B (10), along with four known dihydrochalcones, phloretin (1), phlorizin (2), asebogenin (3), and asebotin (4), were isolated from the leaves of Pieris japonica. Their structures were elucidated on the basis of spectroscopic analysis including HMQC, HMBC, NOESY, and X-ray crystal diffraction. Compounds 1, 3-5, and 7-10 inhibited the proliferation of murine B cells and compounds 5 and 10 inhibited the proliferation of murine T cells in vitro significantly. PMID- 15787443 TI - Hemiterpene glucosides with anti-platelet aggregation activities from Ilex pubescens. AB - Two new hemiterpene glucosides named pubescenosides A and B were isolated from the root of Ilex pubescens. Their structures were elucidated on the basis of spectroscopic and chemical evidence as 2-(trans-caffeoyloxy)methyl-3-hydroxy-1 butene-4-O-beta-D-glucopyranoside and 2-hydroxymethyl-3-caffeoyloxy-1-butene-4-O beta-D-glucopyranoside, respectively. Pharmacological investigation on pubescenosides A and B indicated that both possess potent anti-platelet aggregation activities. PMID- 15787444 TI - Botryane metabolites from the fungus Geniculosporium sp. isolated from the marine red alga Polysiphonia. AB - Eleven new botryane metabolites (1-11) were isolated together with four known cytochalasins (12-15) from the mitosporic fungus Geniculosporium sp., which is associated with the red alga Polysiphonia sp. The structures of 1-11 differ from known botryanes in substitution pattern, degree of saturation, and altered sites of oxidation, alkylation, unsaturation, etc. They were determined by spectroscopic methods (mainly extensive 1D and 2D NMR experiments and mass spectral measurements) and X-ray single-crystal analysis. The herbicidal, antifungal, and antibacterial activities of these new natural products were evaluated. PMID- 15787445 TI - Tryptamine-derived amides and alkaloids from the seeds of Annona atemoya. AB - A series of N-fatty acyl tryptamines, including a mixture of N nonadecanoyltryptamine (1), N-behenoyltryptamine (2), N-lignoceroyltryptamine (3), N-cerotoyltryptamine (4), and N-octacosanoyl tryptamine (5), and a mixture of N-tricosanoyl-4,5-dihydroxytryptamine (6), N-lignoceroyl-4,5 dihydroxytryptamine (7), N-pentacosanoyl-4,5-dihydroxytryptamine (8), and N heptacosanoyl-4,5-dihydroxytryptamine (9), along with two alkaloids, atemoine (10) and cleistopholine, were isolated from the EtOAc extract of seeds of Annona atemoya. The structures of the new compounds, 1 and 5-10, were determined on the basis of spectral evidence. PMID- 15787446 TI - Cerylimycins [corrected] A and B: antibacterial triterpenes from the new species Tricholoma sp. AU1. AB - To new triterpenes, trichomycins A (1) and B (2), were purified from the new species Tricholoma sp. AU1 by activity-guided fractionation following their antibacterial activity. The two compounds were found to have a hitherto unreported triterpenoid skeleton. The structures and relative stereochemistry of 1 and 2 were determined through extensive 2D NMR spectroscopy, while the inhibitory activity of 1 and 2 against two Gram-positive and two Gram-negative bacteria and a mammalian cell line was determined. PMID- 15787447 TI - Acylperoxylated and seco-mexicanolides from stems of Khaya anthotheca. AB - The stems of Khaya anthotheca yielded two new limonoids, which were identified on the basis of spectroscopic analysis as 1alpha,8alpha-oxido-3beta-acetoxy-2alpha acylperoxy-1alpha,14alpha-dihydroxy-[3.3.1(10,2)]-bicyclomeliac-7,19-olide (3) and 3-acetoxy-8,14-dien-8,30-seco-khayalactone (4). Methyl 1alpha,2beta,3alpha,6,8alpha,14beta-hexahydroxy-[4.2.1(10,30).1(1,4)] tricyclomeliac-7-oate, scopoletin, and 3-O-beta-D-glucopyranosylsitosterol were also isolated. The limonoids were of little value to clarify the basis of the nonresistance against Hypsipyla grandella. PMID- 15787448 TI - Cytotoxic flavanones of Schizolaena hystrix from the Madagascar rainforest. AB - Bioassay-guided fractionation of an ethanol extract of a Madagascar collection of Schizolaena hystrix afforded three new flavanones, schizolaenone A (1), schizolaenone B (2), and 4'-O-methylbonannione A (3), as well as three known flavanones, nymphaeol A, bonannione A, and macarangaflavanone B, and the flavanol bonanniol A. The structures of compounds 1-3 were determined by various one- and two-dimensional NMR techniques. All of the isolates were tested for cytotoxicity against the A2780 human ovarian cancer cell line. Nymphaeol A (IC(50) = 5.5 microg/mL) exhibited the greatest cytotoxicity, while the other flavanones were found to be only weakly active. PMID- 15787449 TI - Glycoyessotoxin a, a new yessotoxin derivative from cultures of Protoceratium reticulatum. AB - The dinoflagellate Protoceratium reticulatum produces toxins of the yessotoxin group currently included in the diarrhetic shellfish poisoning class. In this paper we report on the isolation and structural elucidation of a 32-arabinoside of yessotoxin, G-YTXA (2), obtained from laboratory cultures of P. reticulatum (strain GG1AM) that possesses a pentose unit, beta-arabinofuranose, as a side chain. PMID- 15787450 TI - Cytotoxic flavonoids from Platymiscium floribundum. AB - Two new isoflavonoids, 7-hydroxy-6,4'-dimethoxy-isoflavonequinone (1) and 2' hydroxy-6,4',6' ',4' ''-tetramethoxy-[7-O-7' ']-bisisoflavone (2), and seven other known flavonoids, 3-hydroxy-9-methoxypterocarpan (medicarpin), 3,10 dihydroxy-9-methoxypterocarpan, 3,9-dimethoxypterocarpan (homopterocarpin) (3), 2,3,9-trimethoxypterocarpan (4), 3,4-dihydroxy-9-methoxypterocarpan (vesticarpan) (5), 2',4,4'-trihydroxychalcone (isoliquiritigenin), and 7,4'-dihydroxyflavanone (liquiritigenin) (6), were isolated from the heartwood of Platymiscium floribundum. The structures of compounds 1 and 2 were established by spectroscopic methods. Compounds 3-6 showed cytotoxic activity when evaluated against five human cancer cell lines in vitro. PMID- 15787451 TI - Bisindigotin, a TCDD antagonist from the Chinese medicinal herb Isatis indigotica. AB - A new indigoid derivative, bisindigotin (1), with 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD)-antagonistic activity was isolated from the ethanol extract of the Chinese medicinal herb Isatis indigotica. Its structure was determined by spectroscopic methods. In the human HepG2 hepatoma cell model, 1 (50 nM to 2 microM) was found to dose-dependently inhibit TCDD-induced ethoxyresorufin O deethylase (EROD) activity. PMID- 15787452 TI - Acetylated aporphine alkaloids from Lysichiton camtschatcense. AB - Two N,O-diacetylaporphine alkaloids, N,O-diacetylnoroliveroline (1) and N,O diacetyl-(-)-nornuciferidine (2), have been isolated and characterized from the rhizomes of Lysichiton camtschatcense. Their structures were determined by spectroscopic data analysis. DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals were scavenged by compounds 1 and 2, but with weak activity. PMID- 15787453 TI - Anthecotulide: purification, analytical data, absence from chamomile preparations, stability and reactivity, and anti-infective testing. AB - The sesquiterpene lactone anthecotulide (1) was extracted from Anthemis cotula in 0.7% w/w yield. A pure standard was prepared by preparative TLC and used for quantitative determination of 1 with a TLC scanner. The constitution of 1 was corroborated and the double-bond configuration determined. It was stable at room temparature in air and reacted very slowly with glutathione. It was not detectable in 34 chamomile (Matricaria recutita) preparations and did not show antibacterial activity against a range of clinically important strains at a concentration of <10 microg/mL. PMID- 15787454 TI - Communiols E-H: new polyketide metabolites from the coprophilous fungus Podospora communis. AB - Communiols E-H (1-4), four new polyketide-derived natural products containing furanocyclopentane, furanocyclopentene, cyclopentene, or gamma-lactone moieties, have been isolated from two geographically distinct isolates of the coprophilous fungus Podospora communis. The structures of these compounds were determined by analysis of NMR and MS data. PMID- 15787455 TI - Eremophilane-type sesquiterpene derivatives from Senecio aegyptius var. discoideus. AB - Investigation of a CH(2)Cl(2) extract of the aerial parts of Senecio aegyptius var. discoideus afforded nine eremophilane compounds, of which six are new (1-6), namely, 1beta-hydroxy-8alphaH-eremophil-7(11),9-dien-8beta,12-olide (1), 1beta,8alpha-dihydroxyeremophil-7(11),9-dien-8beta,12-olide (2), 1beta-hydroxy 8alpha-methoxyeremophil-7(11),9-dien-8beta,12-olide (3), 1-oxo-8alpha-methoxy 10alphaH-eremophil-7(11)-en-8beta,12-lactam (4), 1beta,10beta-epoxy-8alpha hydroxyeremophil-7(11)-en-8beta,12-olide (5), and 1beta,10beta-epoxy-8alpha methoxyeremophil-7(11)-en-8beta,12-olide (6). The structures of 1-6 were elucidated by spectroscopic methods and by comparison with literature data. The antibacterial activity of the isolated compounds was tested against Bacillus cereus and a Serratia sp. PMID- 15787456 TI - Hopane-type saponins from the seeds of Glinus lotoides. AB - Four new hopane-type saponins, glinusides F, G, H, and I (1-4), and the known succulentoside B (5), as well as the two known flavones 5,7,4'-trihydroxyflavone 6,8-di-C-glucoside (vicenin-2) and 5,7,4'-trihydroxyflavone-8-C-sophoroside (vitexin-2' '-O-glucoside), were isolated from the seeds of Glinus lotoides growing in Ethiopia. On the basis of the spectroscopic data analysis, including 2D NMR and HRESIMS, the new structures were characterized as 3beta-O-beta-D xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta-O-beta-D-xylopyranosyl-22 hydroxyhopane (1), 3beta-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl 6alpha,16beta-dihydroxy-22-O-alpha-L-rhamnopyranosylhopane (2), 3beta-O-alpha-L rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta hydroxy-22-O-alpha-L-rhamnopyranosylhopane (3), and 3beta-O-alpha-L rhamnopyranosyl-(1-->2)-beta-d-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta O-beta-D-xylopyranosyl-22-hopane (4). PMID- 15787457 TI - Sesquiterpenoids and plasmin-inhibitory flavonoids from Blumea balsamifera. AB - Two new sesquiterpenoid esters (1 and 2) were isolated from the extract of Blumea balsamifera, a tropical Compositae plant. Compound 2 proved to be weakly cytotoxic when tested against Jurkat human T-cell leukemia cells. Nine known flavonoids, of which two showed plasmin-inhibitory activity, were also isolated. PMID- 15787458 TI - A cytotoxic carotenoid from the marine sponge Prianos osiros. AB - Investigations of the marine sponge Prianos osiros, collected in Pohnpei, gave a new cytotoxic acetylenic carotenoid, (3R,3'R,5S)-3,3',5,19'-tetrahydroxy-7',8' didehydro-gamma,epsilon-carotene-8-one. The absolute configuration of this carotenoid was solved by interpretation of IR, MS, and 2D NMR spectra and application of the modified Mosher's method. Compound 1 is cytotoxic toward cultured human colon tumor cells, HCT 116 (IC(50) 4.38 microg/mL). PMID- 15787459 TI - 4-Hydroxybenzoyl derivative from the aqueous extract of the hydroid Campanularia sp. AB - A new compound, N-(4-guanidinobutyl)-2-(4-hydroxyphenyl)-2-oxo-acetamide (1) was isolated from the aqueous extract of the hydroid Campanularia sp. Its structure was elucidated using NMR spectroscopic techniques and mass spectrometric analysis. The most stable conformation was determined using molecular modeling and the results of a NOESY experiment. Although compound 1 shows structural similarities to some highly potent histone deacetylase inhibitors (HDACi), e.g., suberoylanilide hydroxamic acid (SAHA) (2) and trichostatin A (TSA) (3), it does not inhibit the growth of ARP-1 cells at 100 microM concentration, a significant indication that it has no inhibitory activity to HDACs. PMID- 15787460 TI - Cytotoxic xanthones from Cudrania tricuspidata. AB - The new isoprenylated tetrahydroxyxanthone, 2,3,6,8-tetrahydroxy-1-(3-methylbut-2 enyl)-5-(2-methylbut-3-en-2-yl)-9H-xanthen-9-one (1), was isolated from the root bark of Cudrania tricuspidata together with macluraxanthone B (2) and cudraxanthone L (3), which were fully characterized by NMR spectroscopic and X ray crystallographic analyses. PMID- 15787461 TI - Ambigol C and 2,4-dichlorobenzoic acid, natural products produced by the terrestrial cyanobacterium Fischerella ambigua. AB - The new natural products 3,5-bis(2,4-dichlorophenoxy)-2,6-dichlorophenol (ambigol C, 1), a highly chlorinated aromatic compound, and 2,4-dichlorobenzoic acid (2) were isolated from the terrestrial cyanobacterium Fischerella ambigua together with the known compounds ambigol A (3) and tjipanazole D (4). All structures were secured by extensive spectroscopic analysis (1D and 2D NMR, MS, UV, IR). Ambigol C has moderate activity against Trypanosoma rhodesiense. PMID- 15787462 TI - Brasilibactin A, a cytotoxic compound from actinomycete Nocardia brasiliensis. AB - A new cytotoxic compound, brasilibactin A (1), has been isolated from the actinomycete Nocardia brasiliensis IFM 0995, and the structure was elucidated on the basis of spectroscopic data and chemical means. PMID- 15787463 TI - Umbelactonyl cinnamate derivatives from Crypteronia paniculata that mediate DNA strand scission. AB - As part of an initiative to discover functional natural product analogues of bleomycin guided by the use of the COMPARE algorithm, a CH(2)Cl(2)-MeOH extract prepared from Crypteronia paniculata was found to exhibit relaxation of supercoiled pSP64 DNA in the presence of Cu(2+). Bioassay-guided fractionation employing a DNA strand scission assay resulted in the isolation of three novel DNA cleavage agents. Their structures were elucidated as umbelactonyl cinnamate derivatives 1-3 through their NMR and MS spectral data analyses. This is the first example of the isolation and structural characterization of naturally occurring umbelactonyl cinnamate derivatives. Compound 1 exhibited strong Cu(2+) dependent relaxation of supercoiled pSP64 DNA, while compounds 2 and 3 had only weak DNA cleavage activity. PMID- 15787464 TI - Daucane sesquiterpenes from Ferula hermonis. AB - The dried roots of Ferula hermonis yielded three new daucanes, (1R,4R)-4 hydroxydauca-7-ene-6-one (1), (1R,4R)-4-hydroxydauca-7-ene-6,9-dione (2), and (1R,3S,8S)-3-ethoxy-8-angeloyloxydauca-4-en-9-one (3), together with the three known sesquiterpenes, ferutinin, teferidin, and (+)-alpha-bisabolol. The structures of compounds 1-3 were elucidated on the basis of spectroscopic evidence. The effect of these compounds on the proliferation of estrogen dependent MCF-7 cells was evaluated, and it was found that compounds 1 and 3 exhibited proliferative activity, whereas 2 showed an antiproliferative effect. PMID- 15787466 TI - End-cap exchange of rotaxane by the Tsuji-Trost allylation reaction. AB - [reaction: see text] Rotaxanes possessing a cinnamyl ester group at the axle terminal were prepared. The terminal end-cap was modified with a bulky malonate ester in excellent yield by the Tsuji-Trost allylation reaction, which was carried out in the presence of a palladium catalyst. PMID- 15787465 TI - Exploring the chemistry of uncultivated bacterial symbionts: antitumor polyketides of the pederin family. AB - Symbiotic bacteria have long been proposed as being responsible for the production of numerous natural products isolated from invertebrate animals. However, systematic studies of invertebrate-symbiont associations are usually associated with serious technical challenges, such as the general resistance of symbionts to culturing attempts and the complexity of many microbial consortia. Herein an overview is provided on the culture-independent, metagenomic strategies recently employed by our group to contribute to a better understanding of natural product symbiosis. Using terrestrial Paederus spp. beetles and the marine sponge Theonella swinhoei as model animals, the putative genes responsible for the production of pederin-type antitumor polyketides have been isolated. In Paederus fuscipes, which uses pederin for chemical defense, these genes belong to an as yet unculturable symbiont closely related to Pseudomonas aeruginosa. To study the extremely complex association of T. swinhoei and its multispecies bacterial consortium, we used a phylogenetic approach that allowed the isolation of onnamide/theopederin polyketide synthase genes from an uncultured sponge symbiont. Analysis of the biosynthesis genes provided unexpected insights into a possible evolution of pederin-type pathways. Besides revealing new facets of invertebrate chemical ecology, these first gene clusters from uncultivated symbiotic producers suggest possible biotechnological strategies to solve the supply problem associated with the development of most marine drug candidates. PMID- 15787468 TI - Chiral trialkanolamine-based hemicryptophanes: synthesis and oxovanadium complex. AB - [reaction: see text] A novel class of chiral hemicryptophane hosts has been synthesized in diastereoisomerically pure form, namely, M-(R,R,R)-1a/P-(S,S,S)-1a and M-(S,S,S)-1b/P-(R,R,R)-1b. The C3-symmetrical precursor 9 was prepared, using either (R)- or (S)-glycidyl nosylate, repectively, as the chiral pool reactant and subsequently cyclized (trimerized) in the presence of Sc(OTf)3. The four stereoisomers were fully characterized and displayed two pairs of mirror-image CD spectra, which were used to determine their absolute configuration. The formation of the oxovanadium(V) complex of hemicryptophane 1a is also reported. PMID- 15787467 TI - Bridged peptide macrocycles as ligands for PDZ domain proteins. AB - [reaction: see text] Conformationally constrained side chain-bridged cyclic peptides were prepared using bis-carboxylic acid ring spacers. These macrocyles were designed to inhibit protein-protein interactions mediated by the third PDZ domain (PDZ3) of a mammalian neuronal protein, PSD-95. Isothermal titration calorimetry (ITC) experiments measured dissociation constants in the low micromolar range. For each compound, the change in entropy (TdeltaS) of binding either is comparable in magnitude to the enthalpy change (deltaH) or is the predominant driving force for association. PMID- 15787469 TI - One-pot multistep synthesis of 4-acetoxy-2-amino-3-arylbenzofurans from 1-aryl-2 nitroethylenes and cyclohexane-1,3-diones. AB - [reaction: see text] A novel method for synthesizing 4-acetoxy-2-amino-3 arylbenzofurans (4) from 1-aryl-2-nitroethylenes (1) and cyclohexane-1,3-diones (2) is described. The method features one-pot operation of a solution of 1 and 2 in THF with catalytic Et3N (rt, 12 h) followed with Ac2O, Et3N, and DMAP (rt, 5 h), although the process consists of 13 elementary reactions. PMID- 15787470 TI - Total synthesis of (+/-)-herbindole B and (+/-)-cis-trikentrin B. AB - [reaction: see text] Herbindole B and cis-trikentrin B are naturally occurring indoles having the unusual and synthetically challenging pattern of carbon substitution at the 4-7 and 5-7 positions, respectively, with no substitution at the 1-3 positions. The total syntheses of these polyalkylated indoles have been achieved in 19 and 12 steps, respectively. The synthesis of herbindole B relies on two iterations of a quinine monoimine Diels-Alder reaction, while cis trikentrin B uses a single cycloaddition of a suitable quinone monoimine. Indolization of the adducts provides suitably substituted benzopyrrole nuclei for elaboration to the natural products. PMID- 15787471 TI - Iron-catalyzed cross-coupling of alkenyl sulfides with Grignard reagents. AB - [reaction: see text] The iron-catalyzed cross-coupling reaction of alkenyl sulfides with Grignard reagents is described. While the cross-coupling proceeds efficiently at alkenyl-S bonds, almost no cross-coupling takes place at aryl-S bonds, attesting to a unique selectivity of iron catalysis. The beneficial effect of potentially coordinating 2-pyrimidyl group on sulfur is also described. PMID- 15787472 TI - Alkylation of magnesium sulfinates: a direct transformation of functionalized aromatic/heteroaromatic halides into sulfones. AB - [reaction: see text] Sulfinate alkylation is one of the conventional methods for sulfone synthesis. The alkylation of magnesium sulfinates, which are easily accessible via reactions of organomagnesium intermediates with sulfur dioxide, provides a convenient route for sulfone preparation. In this communication, we report a preliminary study of the alkylation of arylmagnesium sulfinates. An application of this reaction to directly transform functionalized aromatic/heteroaromatic halides into sulfones is also described. PMID- 15787473 TI - Total synthesis of (+)-trans-195A. AB - [reaction: see text] The first enantioselective synthesis of (+)-trans-195A is described. The structure has been constructed by ring-rearrangement metathesis (RRM) and zirconium-mediated Negishi-coupling, used for the first time to prepare 6,6-membered heterocycles, as key steps. By comparison of the synthesized material with the isolated natural product, the absolute configuration of natural trans-195A was determined to be (2R,4aS,5R,8aS)-(-). PMID- 15787474 TI - Mono- and dialkylations of pyrrole at C2 and C5 positions by nucleophilic substitution reaction in ionic liquid. AB - [reaction: see text] A novel ionic liquid methodology for pyrrole C-alkylation is described. The pyrrole alkylation is achieved with various simple alkyl halides and mesylates selectively at C2 and C5 positions in good yields with minimal byproducts under relatively mild conditions in various ionic liquids. 2-(3 Phenylpropyl)pyrrole (2a) was synthesized from pyrrole and 1-bromo-3 phenylpropane in a mixture solvent system, [bmim][SbF6] and CH3CN, in 81% yield at 115 degrees C for 44 h with 5% yield of dialkylated compound 3a. PMID- 15787475 TI - A new synthesis of allyl sulfoxides via nucleophilic addition of sulfinyl carbanions to group 6 Fischer carbene complexes. AB - [reaction: see text] A novel synthesis of allyl sulfoxides has been developed. Primary alpha-lithiosulfinyl carbanions react with group 6 Fischer carbene complexes to give allyl sulfoxides as products. The Fischer carbene complex experiments involve a 1,2-addition of two molecules of sulfinyl carbanion to give an intermediate that, after a beta-elimination, furnishes the mentioned product. PMID- 15787476 TI - Highly enantioselective syntheses of heterocycles via intramolecular Ir-catalyzed allylic amination and etherification. AB - [reaction: see text] Enantioselective Ir-catalyzed intramolecular allylic aminations and etherifications are described. Up to 97% ee was achieved using catalysts prepared by in situ activation of mixtures of phosphorus amidites and [Ir(COD)Cl]2. Sequential aminations of bis-allylic carbonates, involving an inter followed by an intramolecular reaction, gave trans-N-benzyl-2,5 divinylpyrrolidine and trans-N-benzyl-2,6-divinylpiperidine with > or = 99% ee. New phosphorus amidites as well as improved conditions for intermolecular aminations are reported. PMID- 15787477 TI - Asymmetric total synthesis of rollicosin. AB - [structure: see text] The first total synthesis of rollicosin, a member of a rare subgroup of Annonaceous acetogenins containing two terminal gamma-lactones, is reported. The approach features a highly regio- and stereoselective tandem ring closing/cross-metathesis reaction for construction of the east-wing lactone and incorporation of the alkyl spacer. Establishment of the C4 stereocenter and addition of the west-wing lactone were achieved by Sharpless asymmetric dihydroxylation and enolate alkylation. PMID- 15787478 TI - Cycloruthenated primary and secondary amines as efficient catalyst precursors for asymmetric transfer hydrogenation. AB - [reaction: see text] Ruthenacycles obtained by cyclometalation of enantiopure aromatic primary or secondary amines with [(eta6-benzene)RuCl2]2 or with [(eta6-p cymene)RuCl2]2 are efficient catalysts for asymmetric transfer hydrogenation (TOF up to 190 h(-1) at room temperature). Enantioselectivities in the transfer hydrogenation of acetophenone ranged from 38% to 89%. It is possible to prepare the catalysts in situ, which allows the use of high throughput experimentation. PMID- 15787479 TI - Reversal of chemoselectivity in Diels-Alder reaction with alpha,beta-unsaturated aldehydes and ketones catalyzed by Bronsted acid or Lewis acid. AB - [reaction: see text] High chemoselectivity was observed in the Diels-Alder reaction of alpha,beta-unsaturated aldehyde and alpha,beta-unsaturated ketone with cyclopentadiene. Using Tf2NH as catalyst, the reaction gave Diels-Alder adduct derived from alpha,beta-unsaturated ketone as a major product. On the other hand, bulky Lewis acid, B(C6F5)3, gave mainly the cycloadduct of alpha,beta unsaturated aldehyde and cyclopentadiene. PMID- 15787480 TI - Catalytic enantioselective synthesis of quaternary all-carbon stereogenic centers. Preparation of alpha,alpha'-disubstituted beta,gamma-unsaturated esters through Cu-catalyzed asymmetric allylic alkylations. AB - [reaction: see text] Peptide-based chiral ligands, readily prepared from commercially available materials, are used to promote Cu-catalyzed asymmetric allylic alkylations of alpha,beta-unsaturated esters bearing a gamma-phosphate with various alkylzinc reagents. These transformations lead to the formation of alpha,alpha'-dialkyl-beta,gamma-unsaturated esters in high yields as well as high regio- (re) and enantioselectivities (ee). PMID- 15787481 TI - Basicity of some phosphines in THF. AB - [reaction: see text] The reaction of several phosphines with an acidic indicator gives both ion pairs and free ions. The value obtained for the pKa of tribenzylphosphine is shown to be reasonable by MO computations. An important limitation is demonstrated for the Fuoss equation of dissociation of ion pairs. PMID- 15787482 TI - Lancifodilactone F: a novel nortriterpenoid possessing a unique skeleton from Schisandra lancifolia and its anti-HIV activity. AB - [structures: see text] Lancifodilactone F (1), possessing an unprecedented rearranged pentanortriterpenoid backbone derived from cycloartane, was isolated from the leaves and stems of Schisandra lancifolia (Rehd. et Wils) A. C. Smith. Its structure was established by comprehensive NMR and MS spectroscopic analysis, coupled with single-crystal X-ray experiment. Compound 1 exerted minimal cytotoxicity against C8166 cells (CC50 > 200 microg/mL) and showed anti-HIV activity with EC50 = 20.69 +/- 3.31 microg/mL and a selectivity index > 6.62. PMID- 15787483 TI - Electrophilic fluorodesilylation of allenylmethylsilanes: a novel entry to 2 fluoro-1,3-dienes. AB - [reaction: see text] Various fluorodienes were prepared by treatment of the corresponding allenylmethylsilanes with Selectfluor. This is the first route to these compounds not based on the use of a fluorinated building block. The reaction allows the preparation of 2-fluoro-1,3-dienes with several substitution patterns, including di- and trisubstituted compounds. PMID- 15787485 TI - Concise and enantioselective synthesis of the aminocyclitol core of hygromycin A. AB - [reaction: see text] Stereoselective aminohydroxylation and dihydroxylation using osmium(VIII) oxidants enabled the short and efficient synthesis of the aminocyclitol core of hygromycin A. In addition to allowing the selective introduction of the heteroatoms N and O, the use of osmium (via an osmate ester) as a protecting group for a 1,2-glycol is also reported. This tactic allowed efficient differentiation of otherwise equivalent hydroxyl groups and allowed us to complete the synthesis in short order (14 steps) and excellent overall yield (12%). PMID- 15787484 TI - Novel 3',6'-anhydro and N12,N13-bridged glycosylated fluoroindolo[2,3 a]carbazoles as topoisomerase I inhibitors. Fluorine as a leaving group from sp3 carbon. AB - [reaction: see text] Both 6'- and 4'-fluoro-glycosylated indolo[2,3-a]carbazoles are substrates for base-induced loss of fluorine as a leaving group from sp3 carbon. In the case of alpha-N-glycosylated substrate 3, loss of fluorine from the 6'-position leads to 3,6-anhydroglucose analogue 1. A novel N12,N13-bridged sugar analogue 2 results from loss of 4'-fluorine from beta-N-glycosylated analogue 4. Both analogues 1 and 2 display topo I inhibitory potencies similar to camptothecin. PMID- 15787486 TI - Building blocks for self-assembled porphyrinic photonic wires. AB - [structure: see text] Based on the high affinity of phenanthroline-strapped porphyrins for imidazoles, building blocks for self-assembled, linear porphyrin architectures have been designed. Their synthesis is reported, and the assembly principle is illustrated by the formation of the shortest possible scaffold. Only one type of assembly is observed, and the stepwise energy transfer from the boron dipyrrylmethane (BODIPY) input to the free base output is highly efficient. PMID- 15787487 TI - Design and synthesis of a neutral fluorescent macrocyclic receptor for the recognition of urea in chloroform. AB - [structure: see text] An artificial macrocyclic fluorescent receptor 1 has been designed and synthesized for the recognition of urea. 1 shows significant fluorescence quenching on complexation with urea and thiourea in chloroform and thus may be used as a synthetic fluorescent molecular sensor for their determination in a nondegradative way. PMID- 15787488 TI - Positive dendritic effects on the electron-donating potencies of poly(propylene imine) dendrimers. AB - [structure: see text] Two series of poly(propylene imine), PPI, dendrimers terminated with a redox-active donor, 4-dimethylaminobenzyl (4-DMAB), including their respective nondendronized model compounds, are reported. In these two series, a positive dendritic effect was observed for the formation of charge transfer (CT) complexes between the dendrimers and 7,7,8,8 tetracyanoquinodimethane (TCNQ). However, the nondendronized compounds did not form CT complexes with TCNQ, even though their redox potentials are similar to those of the 4-DMAB units attached to the dendrimers. PMID- 15787489 TI - Structure, conformation, and dynamic processes of the stereolabile atropisomers of hindered terphenyl hydrocarbons. AB - [reaction: see text] Ortho-substituted p-terphenyl hydrocarbons exist as trans and cis atropisomers that were identified by low-temperature NMR spectroscopy. The interconversion barriers increase with the dimensions of the ortho substituents, the experimental values being matched by ab initio calculations. X ray diffraction shows that only the trans atropisomer is present in the solids. Spectra of a tert-butyl derivative in nonequilibrium conditions indicate that the cis is more populated than the trans atropisomer in solution, favored by attractive interactions. PMID- 15787490 TI - Convergent synthesis of internally branched PAMAM dendrimers. AB - [reaction: see text] A series of aliphatic internally branched poly(amido amine) dendrons and dendrimers has been synthesized. The internal branching unit was 1,2 propanediamine and a series of PAMAM-type dendrons of the types AB2, AB4, and AB8 were built. These were anchored on a core molecule containing four carboxylic acid moieties and the 1,2-propanediamine unit resulted in PAMAM dendrimers with 4, 8, 16, and 32 end groups. PMID- 15787491 TI - Stereoselective synthesis of 4-(N-mesylamino)-2,3-unsaturated- alpha-O-glycosides via a new glycal-derived vinyl alpha-N-(mesyl)-aziridine. AB - [reaction: see text] N-Mesyl aziridine 7alpha, a new activated vinyl aziridine derived from d-glucal, has been synthesized by cyclization of trans-N,O dimesylate 6 with t-BuOK in anhydrous benzene. The reaction of 7alpha with alcohols, phenol, and monosaccharides (O-nucleophiles) leads to the corresponding 4-N-(mesylamino)-2,3-unsaturated-O-glycosides and disaccharides through a completely regioselective 1,4-addition process that proceeds with high or complete alpha-stereoselectivity. PMID- 15787492 TI - Total synthesis of (+)-peloruside A. AB - [reaction: see text] A total synthesis of (+)-peloruside A has been successfully achieved. The strategy was highlighted by a late stage aldol coupling of two complex fragments followed by an intramolecular hemi-ketal cyclization, a MOM group participated epoxide ring fragmentation reaction, and a highly selective methylation. This convergent route allows access to rationally designed analogues. PMID- 15787493 TI - The isopropylsulfinyl group: a useful chiral controller for the asymmetric aziridination of sulfinylimines and the organocatalytic allylation of hydrazones. AB - [reaction: see text] A comparative study shows that the isopropylsulfinyl group for which an enantiodivergent and highly diastereoselective approach is reported behaves better than the tert-butylsulfinyl and p-tolylsulfinyl groups, both in terms of reactivity and stereoselectivity in the Corey-Chaykovsky reaction of chiral sulfinylimines and the organocatalytic allylation of acyl hydrazones. PMID- 15787494 TI - Synthesis of epothilones via a silicon-tethered RCM reaction. AB - [reaction: see text] A short synthesis of epothilone B and D is reported. The key step for generating the C12-13-trisubstituted Z-double bond uses a ring-closing metathesis reaction of a disiloxane to form a nine-membered silicon-tethered ring. PMID- 15787495 TI - Photochemistry of 1-isopropylcycloalkyl aryl ketones: ring size effects, medium effects, and asymmetric induction. AB - [reaction: see text] The n = 0, 1, and 2 ketones shown above undergo Yang photocyclization in solution, but only the n = 1 analogues react this way in the solid state. Based on X-ray crystallography, these differences in reactivity are attributed to an unusually large distance for 1,4-hydroxybiradical cyclization in the solid state for the n = 0 and 2 ring systems, which leads to predominant reverse hydrogen transfer (rht). Enantiomeric excesses of up to 99% can be achieved in the case of the n = 1 system through the use of the solid-state ionic chiral auxiliary method of asymmetric synthesis. PMID- 15787496 TI - Aryl- and alkenyllithium preparations and copper-catalyzed reaction between the derived magnesium reagents and the monoacetate of 4-cyclopentene-1,3-diol. AB - [reaction: see text] Aryl- and alkenyllithiums, prepared by halogen-lithium exchange with lithium, exchange with n-(or t-)BuLi, stannane-lithium exchange with n-BuLi, and direct lithiation with n-BuLi, were transformed into magnesium reagents with MgCl2 and subjected to CuCN-catalyzed reaction with the title monoacetate. Except for the halogen-lithium exchange with n-BuLi, the other preparations of the lithium anions were found to be compatible with the CuCN catalyzed reaction to afford S(N)2-type products efficiently. PMID- 15787497 TI - "One-step" alkynylation of adamantyl iodide with silver(I) acetylides. AB - [reaction: see text] Silver(I) acetylides allow one-step alkynylation of adamantyl iodide in yields ranging from 25 to 68%. PMID- 15787498 TI - Expedient construction of the vibsanin E core without the use of protecting groups. AB - [reaction: see text] The tricyclic core of vibsanin E was constructed without the use of a protecting group in six steps. The El Gaied Baylis-Hillman variant was key to allowing the Bronsted acid induced tandem cyclization forming rings B and C in one operation. PMID- 15787499 TI - Ytterbium triflate-promoted tandem one-pot oxidation-Cannizzaro reaction of aryl methyl ketones. AB - [reaction: see text] Ytterbium triflate was shown to be an effective catalyst in promoting the synthesis of either isopropyl esters or free alpha-hydroxy arylacetic acids from substituted aromatic glyoxals and aryl methyl ketones, respectively. The reaction to provide acids starting from differently substituted ketones was carried out by an environmentally friendly method using an aqueous medium as a solvent and giving the adducts in 78-99% yield without any further purification after the usual workup. PMID- 15787501 TI - An aza-Wittig/pi-furan cyclization approach toward the homoerythrina alkaloid (+/ )-selaginoidine. AB - [reaction: see text] A one-pot procedure was developed to efficiently prepare hexahydroindolinones containing a tethered furan ring. Reaction of a furanyl azide with Bu3P delivered the iminophosphorane, which was allowed to react with 1 methyl-(2-oxocyclohexyl)acetic acid to give the desired hexahydroindolinone ring system. Further treatment with trifluoroacetic acid afforded the tetracyclic lactam skeleton found in the alkaloid (+/-)-selaginoidine. PMID- 15787502 TI - Thiazolium-mediated multicomponent reactions: a facile synthesis of 3-aminofuran derivatives. AB - [reaction: see text] A facile synthesis of highly functionalized 3-aminofuran derivatives by the multicomponent reactions of thiazolium salts, aldehydes, and DMAD is described. PMID- 15787503 TI - Enantioselective acylation of secondary alcohols catalyzed by chiral N heterocyclic carbenes. AB - [reaction: see text] The synthetic utility of chiral N-heterocyclic carbenes, which have been used mainly in transition metal-catalyzed reactions as a ligand, was demonstrated by the enantioselective acylation of secondary alcohols. PMID- 15787500 TI - aza-Prins-pinacol approach to 7-azabicyclo[2.2.1]heptanes and ring expansion to [3.2.1]tropanes. AB - [reaction: see text] The 7-azabicyclo[2.2.1]heptane ring system can be rapidly accessed from 5-(1-hydroxyallyl)-2-alkoxy-N-tosylpyrrolidines via an unusual aza Prins-pinacol reaction mediated by Lewis acid. The products can undergo ring expansion to isomeric tropanones. These reactions show promise for a concise entry to biologically relevant azabicyclic targets. PMID- 15787504 TI - Palladium-catalyzed alpha-arylation of sulfoximines. AB - [reaction: see text] Palladium-catalyzed alpha-arylation of N-benzoyl sulfoximine ethyl ester with various aryl bromides leads to alpha-arylated products that can easily be hydrolyzed, giving the corresponding N-protected benzyl phenyl sulfoximines. In this manner, new sulfoximine derivatives are accessible that have so far been difficult to prepare in enantiopure form. PMID- 15787505 TI - Stereoselective synthesis of the octahydronaphthalene unit of integramycin via an intramolecular Diels-Alder reaction. AB - [reaction: see text] The racemic cis-decalin core fragment 30 of integramycin was synthesized by a sequence involving a highly diastereoselective intramolecular Diels-Alder reaction of triene 24. A remarkable switch in stereoselectivity occurred upon changing the dienophile unit of 24 from (Z)- to (E)-geometry. PMID- 15787506 TI - Preparation of a multitopic glycopeptide-oligonucleotide conjugate. AB - [structure: see text] A novel strategy to prepare glycopeptide-oligonucleotide conjugates bearing a glycocluster is reported. The strategy utilizes a cyclodecapeptide scaffold as a key intermediate to anchor the carbohydrate cluster and the oligonucleotide through sequential oxime bond formation. The oligonucleotide glycocluster retains the binding affinity and recognition specificity for the target sequence. Furthermore, the conjugate shows enhanced binding to the specific lectins due to the cooperative effect produced by the carbohydrate cluster. PMID- 15787507 TI - A new entry in catalytic alkynylation of aldehydes and ketones: dual activation of soft nucleophiles and hard electrophiles by an indium(III) catalyst. AB - [reaction: see text] A new entry in catalytic alkynylation of carbonyl compounds was developed in which dual activation of both soft nucleophiles (terminal alkynes) and hard electrophiles (aldehydes and ketones) is achieved using an indium(III) catalyst. Preliminary mechanistic studies using in situ IR and NMR spectroscopic analysis are also discussed. PMID- 15787508 TI - Regio- and stereoselective reductions of gem-difluorinated vinyloxiranes. AB - [reaction: see text] gem-Difluorinated vinyloxiranes are versatile building blocks for the synthesis of fluorinated compounds. Investigations of their reactions with nucleophiles resulted in highly regio- and stereoselective reductions. In their reactions with LiAlH4, hydride reacted at the allylic epoxide carbon to produce homoallylic alcohols exclusively. Moreover, regio- and stereoselective S(N)2' reactions were observed with DIBAL-H and BH3 x THF; the former afforded E allylic alcohols, whereas the latter furnished the corresponding Z isomers with excellent selectivities. PMID- 15787509 TI - Stereochemistry of oxachlorocarbene S(N)i reactions. AB - [reaction: see text] 3-Nortricyclyloxychlorocarbene and trans-4 methylcyclohexyloxychlorocarbene both fragment in hydrocarbon solvents with extensive loss of stereochemical integrity to the corresponding chlorides via competitive and nearly isoenergetic S(N)i-like transition states. PMID- 15787511 TI - Lathyranoic acid A: first secolathyrane diterpenoid in nature from Euphorbia lathyris. AB - [structures: see text] Lathyranoic acid A (1), the first secolathyrane diterpenoid with an unprecedented skeleton, and a new diterpenoid Euphorbia factor L11 (2) were isolated from the seeds of Euphorbia lathyris. Their structures were elucidated by spectroscopic analysis and chemical methods. A biogenetic route involving an enzymatic Baeyer-Villiger oxidation as the key step was postulated for the transformation of 2 to 1 and mimicked by an unusual chemical Baeyer-Villiger oxidation. PMID- 15787510 TI - Isolation of swinholide A and related glycosylated derivatives from two field collections of marine cyanobacteria. AB - [structure: see text] Chemical investigation of two field collections of marine cyanobacteria has led to the discovery of two new cytotoxic natural products, ankaraholides A (2) and B (3), along with the known compound swinholide A (1). Since swinholide-type compounds were previously localized to the heterotrophic bacteria of sponges, these findings raise intriguing questions about their true metabolic source. PMID- 15787512 TI - Mimicking dihydroxy acetone phosphate-utilizing aldolases through organocatalysis: a facile route to carbohydrates and aminosugars. AB - [reaction: see text] A practical and environmentally friendly organocatalytic strategy designed to mimic the DHAP aldolases has been developed and shown to be effective in the preparation of carbohydrates and aminosugars. (S)-Proline and (S)-2-pyrrolidine-tetrazole catalyzed the aldol reaction between dihydroxy acetone variants such as 1,3-dioxan-5-one and 2,2-dimethyl-1,3-dioxan-5-one with aldehydes to give the corresponding polyols in good yields with very high ees. PMID- 15787513 TI - Phosphine-catalyzed synthesis of 1,3-dioxan-4-ylidenes. AB - [reaction: see text] A phosphine-catalyzed reaction of an allenoate with aldehydes furnished (2,6-diaryl-[1,3]dioxan-4-ylidene)-acetates 4 in excellent to moderate yields with complete diastereoselectivity and high E/Z-selectivities. Upon removal of the acetal functionality in this domino reaction product 4, delta hydroxy-beta-ketoester 11 was obtained. The reported vinylphosphonium-based approach provides a new way to achieve a synthesis of delta-hydroxy-beta ketoesters that differs from the classical dianion-based approach. PMID- 15787514 TI - Facile one-pot synthesis of 2,3,5-substituted 1,2,4-oxadiazolines from nitriles in aqueous solution. AB - [reaction: see text] Alkyl/aryl amidoximes, prepared from the corresponding nitriles and N-alkylhydroxylamines, have readily undergone consecutive Michael additions to electron-deficient alkynes and provided highly substituted 1,2,4 oxadiazolines in good yields in homogeneous aqueous solution. PMID- 15787515 TI - Synthesis and stereoselective glycosylation of D- and L-glycero-beta-D-manno heptopyranoses. AB - [reaction: see text] A method for the direct stereocontrolled synthesis of D- and L-glycero-beta-D-manno-heptopyranosides such as those found in the repeating unit of the O-specific polysaccharide from the CNCTC 113/92 LPS (serotype 54) is described. The method relies on the presence of a 4,6-O-benzylidene acetal to effect stereocontrol at the anomeric center; the configuration at C6 (L- or D glycero) is of minimal importance. PMID- 15787516 TI - Novel enantioselective synthesis of 1,3-butadien-2-ylmethanols via tandem alkylbromide-epoxide vinylations using dimethylsulfonium methylide. AB - [reaction: see text] The treatment of chiral trans-disubstituted and trisubstituted 2,3-epoxy-1-bromides with an excess of dimethylsulfonium methylide 1 affords the corresponding 1,3-butadien-2-ylmethanols in good to excellent yields via a double one-carbon homologation. PMID- 15787517 TI - Stereoselective synthesis of alpha-silylamines by the direct addition of silyl anions to activated imines. AB - [reaction: see text] A highly efficient stereoselective synthesis of unusual alpha-silylamines via a direct silyl anion addition reaction is reported. This approach is convergent and avoids any problematic aza-Brook shifts of the anionic intermediates. The use of enantiopure tert-butanesulfinyl imines as the electrophiles affords exceedingly high levels of diastereocontrol for the newly formed stereogenic carbon. PMID- 15787518 TI - Organosilanols as catalysts in asymmetric aryl transfer reactions. AB - [reaction: see text] Various ferrocene-based organosilanols have been synthesized in four steps starting from achiral ferrocene carboxylic acid. Applying these novel planar-chiral ferrocenes as catalysts in asymmetric phenyl transfer reactions to substituted benzaldehydes afforded products with high enantiomeric excesses. The best result (91% ee) was achieved in the addition to p chlorobenzaldehyde with organosilanol 2b, which has a tert-butyl substituent on the oxazoline ring and an isopropyl group on the silanol fragment. PMID- 15787519 TI - Synthesis of the C(1)-C(25) fragment of amphidinol 3: application of the double allylboration reaction for synthesis of 1,5-diols. AB - [structure: see text] A synthesis of the C(1)-C(25) fragment of amphidinol 3 is described. The synthesis features two applications of double allylboration reaction methodology for the highly stereoselective synthesis of 1,5-diol units in the C(1)-C(15) segment. PMID- 15787520 TI - Pd(OH)2/C-mediated selective oxidation of silyl enol ethers by tert butylhydroperoxide, a useful method for the conversion of ketones to alpha,beta enones or beta-silyloxy-alpha,beta-enones. AB - [reaction: see text] Pd(OH)2-catalyzed oxidation of silyl enol ethers by t-BuOOH gives either beta-silyloxy-alpha,beta-enones or alpha,beta-enones in good yields depending on the base used. PMID- 15787521 TI - Enantioselective synthesis of (+)- and (-)-dihydroepiepoformin and (+) epiepoformin. AB - [reaction: see text] The enantioselective synthesis of both enantiomers of dihydroepiepoformin (1) and (+)-epiepoformin (2) was achieved from (p tolylsulfinyl)methyl-p-quinols (SR)- or (SS)-3 and (4R,SR)-4, respectively. Key features include the stereocontrolled conjugate addition of AlMe3 to p-quinol 3 and retrocondensation to the ketone functionality, previous to oxidation of the beta-hydroxy sulfoxide moiety of advanced intermediates to the corresponding sulfone. PMID- 15787522 TI - Highly efficient approach to orthogonally protected (2S,4R)- and (2S,4S)-4 hydroxyornithine. AB - [reaction: see text] A concise stereoselective approach to both orthogonally protected (2S,4R)- and (2S,4S)-4-hydroxyornithine, key constituents of the biphenomycin- and clavalanine-type antibiotics, respectively, has been developed. The approach is based on bis(oxazoline) copper(II)-complex-catalyzed diastereoselective Henry reactions of nitromethane with the homoserine-derived aldehyde 6. The synthesis of this versatile chiral building block has been markedly improved. PMID- 15787523 TI - Direct conversion of N-methoxy-N-methylamides (Weinreb amides) to ketones via a nonclassical Wittig reaction. AB - [reaction: see text] N-Methoxy-N-methylamides (Weinreb amides) are converted efficiently into ketones by reaction with alkylidenetriphenylphosphoranes and in situ hydrolysis of the product. PMID- 15787524 TI - Highly exo-selective and enantioselective cycloaddition reactions of nitrones catalyzed by a chiral binaphthyldiimine-Ni(II) complex. AB - [reaction: see text] Significant levels of exo-selectivity (exo:endo = >99:1 to 86:14) and enantioselectivity (95-82% ee) were obtained in the 1,3-dipolar cycloadditions of a number of nitrones with 3-(2-alkenoyl)-2-thiazolidinethiones, using the chiral binaphthyldiimine-Ni(II) complex (5-20 mol %), which was easily prepared form N,N'-bis(3,5-dichrolo-2-hydroxybenzylidene)-1,1'-binaphthyl-2,2' diamine and Ni(ClO4)2 x 6H2O in CHCl3 in the presence of 4 A molecular sieves, as a chiral Lewis acid catalyst. PMID- 15787528 TI - Orthogonally protected lanthionines: synthesis and use for the solid-phase synthesis of an analogue of nisin ring C. AB - [structure: see text] Lanthionine, a thioether analogue of cystine, is a key component of the lantibiotics, a family of modified peptides bearing multiple thioether bridges resulting from posttranslational modifications between side chains. It is also used as a conformational constraint in medicinally active peptides. We have explored two synthetic routes to give lanthionine, orthogonally protected with Alloc/allyl and Fmoc groups. One route utilized a carbamate protected iodoalanine that yielded a mixture of diastereoisomers, and one utilized a trityl-protected iodoalanine, formed via a Mitsunobu reaction, that gave the single desired lanthionine, in complete regio- and diastereoselectivity. We then used this orthogonally protected lanthionine in the solid-phase synthesis of an analogue of a fragment of nisin containing its ring C. The chemoselective deprotection of the allyl and Alloc groups of the incorporated lanthionine unit was followed by regio- and stereoselective cyclization on resin to give the desired lanthionine-bridged peptide. PMID- 15787529 TI - Nitrile biotransformations for highly enantioselective synthesis of oxiranecarboxamides with tertiary and quaternary stereocenters; efficient chemoenzymatic approaches to enantiopure alpha-methylated serine and isoserine derivatives. AB - [reaction: see text] Biotransformations of a number of differently substituted and configured oxiranecarbonitriles using Rhodococcus sp. AJ270, a microbial whole-cell catalyst that contains nitrile hydratase/amidase, were studied. While almost all trans-configured 3-aryl-2-methyloxiranecarbonitriles and 2,3-dimethyl 3-phenyloxiranecarbonitrile were efficiently hydrated by the action of the less enantioselective nitrile hydratase, the amidase exhibited excellent 2S,3R enantioselectivity against 2-methyl-3-(para-substituted phenyl)oxiranecarboxamides. Under very mild conditions, biotransformations of nitriles provided an efficient and practical synthesis of 2R,3S-(-)-3-aryl-2 methyloxiranecarboxamides, electrophilic epoxides with tertiary and quaternary stereocenters, in excellent yield with enantiomeric excess greater than 99.5%. The synthetic applications of the resulting enantiomerically pure epoxides were demonstrated by convenient and straightforward syntheses of polyfunctionalized chiral molecules possessing a quaternary stereocenter such as R-(+)-2-hydroxy-2 methyl-3-phenylpropionic acid, 2R,3R-(-)-3-amino-2-hydroxy-2-methyl-3 phenylpropionic acid, and 2S,3S-(+)-2-amino-3-hydroxy-2-methyl-3-phenylpropionic acid, employing the regio- and stereospecific epoxide ring opening reactions of 2R,3S-(-)-2-methyl-3-phenyloxiranecarboxamide as the key steps. PMID- 15787527 TI - Crossing conventional boundaries in half a century of research. AB - An overview of the research of the Olah group over five decades--ranging from synthetic reagents and methods to mechanistic-structural studies to new hydrocarbon chemistry and the development of methanol based fuels and hydrocarbon synthesis--is presented. PMID- 15787530 TI - Synthesis of functionalized nitroarylmagnesium halides via an iodine-magnesium exchange. AB - [reaction: see text] Various nitro-substituted aryl and heteroaryl iodides undergo an iodine-magnesium exchange reaction when treated with PhMgCl leading to nitro-containing magnesium organometallics. These Grignard reagents display an excellent stability at temperatures below -40 degrees C and do not undergo electron-transfer reactions. They react as expected with various electrophiles. PMID- 15787531 TI - Cyclopropyl containing fatty acids as mechanistic probes for cytochromes P450. AB - [reaction: see text] The mechanism of aliphatic hydroxylation by cytochromes P450 has been the subject of intense debate with several proposed mechanistic alternatives. Various cyclopropyl containing compounds (radical clocks), which can produce both unrearranged and ring opened products upon oxidation, have been key tools in these investigations. In this study, we introduce several cyclopropyl containing fatty acids 1a-4a with which to probe the mechanism of P450s capable of fatty acid hydroxylation. The probes are shown to be capable of distinguishing radical from cationic intermediates due to the rapid equilibration of isomeric cyclopropyl cations. Ring opening of a radical intermediate in an oxidative transformation is expected to yield a single rearranged alcohol, whereas a cation isomerizes prior to ring opening, leading to two isomeric homoallylic alcohols. Oxidation of these probes by P450(BM3) and P450(BioI) gives results consistent with a radical but not a cationic intermediate in fatty acid hydroxylation by these enzymes. Quantitation of the unrearranged and ring opened products gives remarkably homogeneous rates for oxygen rebound of (2-3) x 10(10) s(-1). The effects of introduction of a cyclopropane ring into a fatty acid upon the regiochemistry of hydroxylation are discussed. PMID- 15787532 TI - Synthesis of the 3-methylene-2-vinyltetrahydropyran unit; the hallmark of the sesquiterpene, hodgsonox. AB - [structure: see text] The synthesis of some simple compounds containing the previously unreported 3-methylene-2-vinyltetrahydropyran system, a unique feature of the liverwort metabolite, hodgsonox is reported. Key features are the creation of an acrolein substituted in the alpha-position with a three-carbon chain bearing a terminal electrophilic site, addition of a vinyl group, and cyclization. PMID- 15787533 TI - Mono-N-methylation of functionalized anilines with alkyl methyl carbonates over NaY faujasites. 4. Kinetics and selectivity. AB - [reaction: see text] In the presence of NaY faujasite as the catalyst, the reaction of bifunctional anilines (1-4: XC6H4NH2; X = OH, CO2H, CH2OH, and CONH2) with methyl alkyl carbonates [MeOCO2R': R' = Me or MeO(CH2)2O(CH2)2] proceeds with a very high mono-N-methyl selectivity (XC6H4NHMe up to 99%), and chemoselectivity as well, with other nucleophilic functions (OH, CO2H, CH2OH, CONH2) fully preserved from alkylation and/or transesterification reactions. Aromatic substituents, however, modify the relative reactivity of amines 1-4: good evidence suggests that, not only steric and electronic effects, but, importantly, direct acid-base interactions between substituents and the catalyst are involved. Weakly acidic groups (OH, CH2OH, CONH2, pKa > or = 10) may help the reaction, while aminobenzoic acids (pKa of 4-5) are the least reactive substrates. The solvent polarity also affects the reaction, which is faster in xylene than in the more polar diglyme. The mono-N-methyl selectivity is explained by the adsorption pattern of reagents within the zeolite pores: a B(Al)2 displacement of the amine on methyl alkyl carbonate should occur aided by the geometric features of the NaY supercavities. Different factors account for the reaction chemoselectivity. Evidence proves that the polarizability of the two nucleophilic terms (NH2 and X groups) of anilines is relevant, although adsorption and confinement phenomena of reagents promoted by the zeolite should also be considered. PMID- 15787534 TI - Conformation of acetate derivatives of sugars and other cyclic alcohols. Crystal structures, NMR studies, and molecular mechanics calculations of acetates. When is the exocyclic C-O bond eclipsed? AB - [reaction: see text] A study of published crystal structures (of O-acetylated sugars for the most part) suggests that the exocyclic C-O bond in acetate esters of cyclic alcohols intrinsically prefers a staggered conformation, although the eclipsed conformation is only slightly less stable. When the acetate is flanked by two equatorial substituents the preferred conformation is close to eclipsed. Over 1500 C-OAc bonds have been analyzed. Diagnostic NMR criteria for torsion angles and MM3 calculations are reported and confirm these conclusions. PMID- 15787535 TI - Regiochemically flexible substitutions of di-, tri-, and tetrahalopyridines: the trialkylsilyl trick. AB - [reactions: see text] 2,4-Difluoropyridine, 2,4-dichloropyridine, 2,4,6 trifluoropyridine, 2,4,6-trichloropyridine and 2,3,4,6-tetrafluoropyridine react with standard nucleophiles exclusively at the 4-position under halogen displacement. However, the regioselectivity can be completely reversed if a trialkylsilyl group is introduced in the 5-position of the 2,4-dihalopyridines or in the 3-position of the 2,4,6-trihalopyridines or 2,3,4,6-tetrahalopyridine. Then only the halogen most remote from the bulky silyl unit (at the 2-position in the case of the 2,4-halopyridines, at the 6-position with the other substrates) gets involved in the exchange process. After removal of the silyl protective group the nucleophile is invariably found to occupy the nitrogen-neighboring position. PMID- 15787536 TI - Preparation of C2-symmetric bicyclo[2.2.2]octa-2,5-diene ligands and their use for rhodium-catalyzed asymmetric 1,4-addition of arylboronic acids. AB - [reaction: see text] C2-symmetric bicyclo[2.2.2]octa-2,5-dienes containing benzyl, phenyl, and substituted phenyl groups at 2 and 5 positions were prepared enantiomerically pure by way of bicyclo[2.2.2]octane-2,5-dione as a key intermediate. These chiral diene ligands were successfully applied to rhodium catalyzed asymmetric 1,4-addition of arylboronic acids to alpha,beta-unsaturated ketones. High enantioselectivity (up to 99% ee) as well as high catalytic activity was observed in the addition to both cyclic and linear substrates. PMID- 15787537 TI - Local aromaticity of [n]acenes, [n]phenacenes, and [n]helicenes (n = 1-9). AB - The local aromaticity of the six-membered rings in three series of benzenoid compounds, namely, the [n]acenes, [n]phenacenes, and [n]helicenes for n = 1-9, has been assessed by means of three probes of local aromaticity based on structural, magnetic, and electron delocalization properties. For [n]acenes our analysis shows that the more reactive inner rings are more aromatic than the outer rings. For [n]phenacenes, all indicators of aromaticity show that the external rings are the most aromatic. From the external to the central ring, the local aromaticity varies in a damped alternate way. The trends for the [n]helicene series are the same as those found for [n]phenacenes. Despite the departure from planarity in [n]helicenes, only a very slight loss of aromaticity is detected in [n]helicenes as compared to the corresponding [n]phenacenes. Finally, because of magnetic couplings between superimposed six-membered rings in the higher members of the [n]helicenes series, we have demonstrated that the NICS indicator of aromaticity artificially increases the local aromaticity of their most external rings. PMID- 15787538 TI - Origin of a large temperature dependence of regioselectivity observed for [2 + 2] photocycloaddition (Paterno-Buchi reaction) of 1,3-dimethylthymine with benzophenone and its derivatives: conformational property of the intermediary triplet 1,4-diradicals. AB - [reaction: see text] The [2 + 2] photochemical additions of 1,3-dimethylthymine (DMT) with benzophenone and its 4,4'-substituted derivatives (BPs), difluoro, di tert-butyl, and dimethoxy benzophenones, have been investigated at a temperature range from -40 to 70 degrees C. The photochemical reactions, which are cycloaddition of the 5-6 double bond of DMT with the carbonyl group of BPs, the so-called the Paterno-Buchi (PB) reaction, reveal largely temperature-dependent regioselectivity. The chemical yields of one series of regioisomers, 2, decrease with the increase of the reaction temperature, but those of another regioisomer series, 3, increase, and thus the ratio of 2/3 is strongly dependent on the temperature (2/3 = ca. 70:30 to 30:70). The temperature dependence of the regioselectivity yields two linear functions in the corresponding Eyring diagrams. The Eyring plot with changed slopes is clearly indicative of the change for the selectivity-determining step in the PB reaction, in which the triplet 1,4 diradicals play a crucial role. Computational studies reveal the conformational equilibrium structures of the triplet 1,4-diradicals, energy barriers between the conformers, and the conjectural equilibrium constants from relative potential energies of the stable conformers. A proposed mechanism can reasonably explain the temperature-dependent regioselectivity and chemical yields of two regioisomers varying with the reaction temperature. PMID- 15787539 TI - 1,3-Dipolar reactions involving corannulene: how does its rim and spoke addition vary? AB - [reaction: see text] Corannulene undergoes 1,3-dipolar reactions with the dipoles, diazomethane, nitrile oxide, and nitrone through its rim and spoke pi bonds; the rim addition yields "one possible" adduct whereas two "regioselective" adducts are formed by spoke addition. Mechanisms of these reactions have been investigated at the B3LYP/6-31G(d) level. Computations show that both rim and spoke additions prefer concerted pathways that lie 2-5 kcal/mol lower in energy than stepwise paths. Stepwise additions can take place in two ways and the activation energies of these two modes differ by 1-2 kcal/mol. A close inspection of the energy profiles reveals that rim addition is more favorable kinetically and thermodynamically than spoke addition in view of lower activation energy and higher exothermicity observed for rim addition. The rim bond of corannulene is more flexible for distortion and also has a stronger double bond (i.e. pi character) than the spoke bond and this facilitates rim addition over spoke addition. Deformation energy analysis also confirms the above through higher deformation in corannulene from the spoke addition when compared to rim addition. In the spoke addition, regio1 reaction is kinetically more favored than regio2 reaction. Attempts to react corannulene in an endohedral fashion have led to the exohedral adduct. Computed activation energies suggest that corannulene acts as a deactivated dipolarophile compared to ethylene. Even more striking is the observation that rim and spoke double bonds in corannulene are part of the local aromatic system but it shows remarkable reactivity compared to benzene despite the loss of aromaticity during the reaction. This is well indicated by computed NICS values. Inclusion of acetonitrile as solvent through the PCM model increases the reaction rate and exothermicity. PMID- 15787540 TI - Organometallic routes to 2,5-dihydroxy-3-(indol-3-yl)benzoquinones. Synthesis of demethylasterriquinone B4. AB - A method has been developed to sequentially add indole-3-mercurials to dichlorinated quinones using palladium catalysis. These reactions can be used in the modular assembly of bis(indol-3-yl)benzoquinones, a significant natural product family. PMID- 15787541 TI - Enantio- and diastereoselective additions to aldehydes using the bifunctional reagent 2-(chloromethyl)-3-(tributylstannyl)propene: application to a synthesis of the C16-C27 segment of bryostatin 1. AB - [reaction: see text] Reactions of the bifunctional allylstannane 2-(chloromethyl) 3-(tributylstannyl)propene with aldehydes have been examined. These generally occur in high yields using Lewis acid promoters and the products can be isolated and purified without incident. Good yields and high enantioselectivities are also realized in catalytic asymmetric allylations (CAA reactions) using the previously described BITIP catalyst system. Protection of the free hydroxyl can be accomplished without cyclization to the derived tetrahydrofuran, although this transformation is also facile. The utility of the incorporated allyl chloride functionality allows for the obvious use of such products in reactions with nucleophiles. Use of these products in a less obvious connective strategy is demonstrated in the synthesis of the C12-C27 segment of bryostatin 1 where a connective, or "lynchpin", double-allylation process was employed. The beta hydroxy allyl chloride obtained from an initial chelation-controlled allylation of aldehyde 16 was converted to allylstannane 19 and applied in a second allylation reaction, thus allowing for a highly convergent synthesis of the bryostatin C ring backbone in a stereoselective fashion. PMID- 15787542 TI - A facile method for the synthesis of thiocarbamates: palladium-catalyzed reaction of disulfide, amine, and carbon monoxide. AB - [reaction: see text] A new method for the synthesis of thiocarbamates has been developed. When dialkyl or diaryl disulfides were allowed to react with secondary amines and carbon monoxide in the presence of a catalytic amount of a palladium complex, the thiocarbamates were obtained in moderate to good yields. In contrast to that of secondary amines, in the reaction of a primary amine, no formation of thiocarbamate was confirmed, but urea was formed in good yield. PMID- 15787543 TI - Synthesis of 5-substituted-1H-indol-2-yl-1H-quinolin-2-ones: a novel class of KDR kinase inhibitors. AB - [reaction: see text] A number of approaches for the synthesis of the 1H-indol-2 yl-1H-quinolin-2-one ring system found in the potent and selective KDR kinase inhibitor 1 are described. The preparation and reaction of trimethylsilylnitrobenzene 26 with 2-methoxy-3-quinolinecarboxaldehyde 28 afforded alcohol 30, which was the key intermediate for the preparation of the target compounds. Conversion of alcohol 30 to either nitroketone 36 or nitrostyrene 45 set the stage for reductive cyclization and the formation of indole 25. The quinolin-2-one functionality was unmasked in the last step to provide compound 1 in 56-60% overall yield from readily available starting materials. PMID- 15787544 TI - Studies on Pd(II)-catalyzed synthesis of (Z)-alpha-haloalkylidene-beta-lactones from cyclocarbonylation of 2-alkynols and the subsequent coupling reactions. AB - [reaction: see text] A good regio- and stereoselectivity was observed for the PdCl2-catalyzed cyclocarbonylation of 2-alkynols with CuCl2 affording (Z)-alpha chloroalkylidene-beta-lactones. The highly optically active (Z)-alpha chloroalkylidene-beta-lactones could be easily prepared from the readily available optically active propargylic alcohols. The Pd(II)-catalyzed cyclocarbonylation of 2-alkynols with CuBr2 was also studied. Although the yields of (Z)-alpha-bromoalkylidene-beta-lactones were low, due to the relatively higher activity of the C-Br bond, the coupling reactions of (Z)-alpha-bromoalkylidene beta-lactones were quite smooth to afford the corresponding products in high yields. A rationale for this reaction is discussed. PMID- 15787545 TI - Furan forming reactions of cis-2-alken-4-yn-1-ones. AB - [reactions: see text] The cis-2-alken-4-yn-1-one, 1-phenyl-cis-2-penten-4-yn-1 one (cis-1), readily dimerizes on treatment with weak acid to give the 1,2 difurylethylenes, trans- and cis-1,2 di(2-(5-phenylfuryl))ethene (trans-1 and cis 2), in 62% and 23% yields, respectively. Trimerization of cis-1 to trans,trans 1,2,3-tri(2-(5-phenylfuryl)cyclopropane (4) occurred as a byproduct of treatment with weak acid. These reactions demonstrate the 2-furylcarbenoid reactivity of cis-2-alken-4-yn-1-ones. PMID- 15787546 TI - Kinetic resolution and chemoenzymatic dynamic kinetic resolution of functionalized gamma-hydroxy amides. AB - [reaction: see text] An efficient kinetic resolution of racemic gamma-hydroxy amides 1 was performed via Pseudomas cepacia lipase (PS-C)-catalyzed transesterification. The enzyme PS-C tolerates both variation in the chain length and different functionalities giving good to high enantioselectivity (E values of up to >250). The combination of enzymatic kinetic resolution with a ruthenium catalyzed racemization led to a dynamic kinetic resolution. The use of 2,4 dimethyl-3-pentanol as a hydrogen source to suppress ketone formation in the dynamic kinetic resolution yields the corresponding acetates in good yield and good to high enantioselectivity (ee's up to 98%). The synthetic utility of this procedure was illustrated by the practical synthesis of the versatile intermediate gamma-lactone (R)-5-methyltetrahydrofuran-2-one. PMID- 15787547 TI - PdCl2-catalyzed efficient transformation of propargylic amines to (E)-alpha chloroalkylidene-beta-lactams. AB - [reaction: see text] The PdCl2-catalyzed cyclocarbonylation reaction of propargylic amines with CuCl2 and benzoquinone afforded (E)-alpha chloroalkylidene-beta-lactams in moderate to good yields. The formation of the corresponding Z-isomers or five-membered products was not observed. The reaction of the readily available optically active propargylic amines provides a convenient synthesis of the corresponding (E)-alpha-chloroalkylidene-beta-lactams with high ee values. The structure and the stereochemistry of the products were established by the X-ray single-crystal diffraction study of (E)-6d and (E)-6e, which indicates that the stereoselectivity in this reaction is different from what was observed with propargylic alcohols. A rationale for this reaction was proposed. PMID- 15787548 TI - A new method for the preparation of 1,5-diynes. Synthesis of (4E,6Z,10Z)-4,6,10 hexadecatrien-1-ol, the pheromone component of the cocoa pod borer moth Conopomorpha cramerella. AB - [reaction: see text] A new method for the synthesis of 1,5-diynes, from the reaction of 1,3-dilithiopropyne and propargyl chlorides, was developed. This new methodology was used to prepare (4E,6Z,10Z)-4,6,10-hexadecatrien-1-ol, one of the pheromone components of the cocoa pod borer moth Conopomorpha cramerella, in 51% overall yield. PMID- 15787549 TI - Photorearrangement of alpha-azoxy ketones and triplet sensitization of azoxy compounds. AB - [reaction: see text] Although some aspects of azoxy group radical chemistry have been investigated, unhindered alpha-azoxy radicals remain poorly understood. Here we report the generation of alpha-azoxy radicals under mild conditions by irradiation of alpha-azoxy ketones 4a,b. These compounds undergo alpha-cleavage to yield radicals 5a,b, whose oxygen atom then recombines with benzoyl radicals to produce presumed intermediate 15. Formal Claisen rearrangement gives alpha benzoyloxyazo compounds 8a,b, which are themselves photolabile, leading to both radical and ionic decomposition. The ESR spectrum of 5a was simulated to extract the isotropic hyperfine splitting constants, which showed its resonance stabilization energy to be exceptionally large. Azoxy compounds have been found for the first time to be good quenchers of triplet excited acetophenone, the main sensitized photoreaction of 7Z in benzene being deoxygenation. While this reaction has been reported previously, it was always in hydrogen atom donating solvents, where chemical sensitization occurred. The principal direct irradiation product of 4bZ and model azoxyalkane 7Z is the E isomer, whose thermal reversion to Z is much faster than that of previously studied analogues. PMID- 15787550 TI - Design and synthesis of asymmetric acyclic phospholipid bolaamphiphiles. AB - [structure: see text] A synthetic route was devised for the generation of asymmetric lipid bolaamphiphiles through the sequential esterification of an alkyldioic acid, bearing distinct terminal protecting groups, with propanylamine and lyso-phosphatidylcholine headgroups. Bolaamphiphile self-assembly was investigated in solvent mixes of varying polarity by nuclear magnetic resonance (NMR) and Fourier transform-infrared (FT-IR) spectroscopy, as well as in water by cryo-high-resolution scanning electron microscopy (cryo-HRSEM). We anticipate that asymmetric lipid bolaamphiphiles will provide facile building blocks for engineering a variety of unique membrane-mimetic structures. PMID- 15787551 TI - Synthesis of pyrazine alcaloids from Botryllus leachi. Diazines 43. AB - [reaction: see text] Regioselective metalation of pyrazines and cross-coupling reactions provides an easy access to botryllazines A and B and to an isomer of botryllazine A with good yields from chloropyrazine. PMID- 15787552 TI - Ketyl-allene cyclizations promoted by samarium(II) iodide. AB - [reaction: see text] Samarium(II) iodide has proven to be an effective reagent for intramolecular reductive coupling reactions. Previous investigations of intramolecular ketyl-olefin coupling reactions provided carbocycles in excellent yield and good diastereoselectivity. This method has been extended to ketyl cyclizations with allenes. Substrates leading to both carbocycles and heterocycles in a selective manner are explored. PMID- 15787553 TI - How important is bishomoaromatic stabilization in determining the relative barrier heights for the degenerate Cope rearrangements of semibullvalene, barbaralane, bullvalene, and dihydrobullvalene? AB - [structure: see text] B3LYP/6-31G* calculations have been used to investigate the origins of the relative barrier heights for the degenerate Cope rearrangements of semibullvalene (1), barbaralane (2), bullvalene (3), and dihydrobullvalene (4). We conclude from our calculations that, of the four transition structures (TSs), that for rearrangement of 1 has the smallest amount of interallylic bonding. Nevertheless, relief of strain in the reactant confers on 1 the lowest barrier to Cope rearrangement. Conjugation between the cyclopropane ring and the pi bond of the etheno bridge in 3 makes the barrier for its Cope rearrangement higher than that for 4 and also contributes to making the barrier for 3 higher than that for 2. However, the relatively low barrier to the Cope rearrangement of 2 is largely due to the TS for this reaction having the largest amount of interallylic bonding of all four TSs. PMID- 15787554 TI - Octanol/water partition of ionic species, including 544 cations. AB - [structure: see text] Partition coefficients of single ions in the octanol/water system (log P(oct)) have been assigned on the (Ph4As+, Ph4P+) = Ph4B- assumption. The log P(oct) values of Cl-, Br-, and I- ions are then used to obtain the partition coefficients of cations from partition coefficients of the neutral combination of anion and cation. Partition coefficients of 544 cations derived from 585 organic salts from the MedChem database have been studied. The contributions of the aliphatic charge N+ and aromatic charge n+ in these cations have been investigated. The results show that the contributions of N+ and n+ in different homologous series are affected by the attached functional groups and are not constant, although the effect of the central cation is constant along any given homologous series. The latter can be accounted for qualitatively and semiquantitatively by the electrostatic theory of ionic solvation of Abraham and Liszi. A number of regression equations have been established between partition coefficients of ions and partition coefficients of the corresponding neutral species. These equations can be used to estimate octanol/water partition coefficients for additional cations to about 0.5 log unit. PMID- 15787555 TI - A straightforward and flexible [4 + 2] route to beta-C-naphthyl-2-deoxy glycosides through tandem hydroboration-ketal reduction: de novo access to C naphthyl-6-fluoro and 6,6-difluoro 2-deoxyglycosides. AB - [reaction: see text] Under standard hydroboration-oxidation conditions, the dihydropyrans 4 underwent a highly stereocontrolled tandem reaction, involving the expected hydration of the double bond together with the reduction of the ketal moiety. This unprecedented transformation gives rise to a short, [4 + 2] based synthetic route to (+/-)-beta-C-naphthyl-2-deoxyglycosides 5, which allows a significant structural and functional diversity at C-6. We thus described the first synthesis of (+/-)-C-aryl-6-fluoro and -6,6-difluoro olivosides, via the allylic mono- and difluorides produced by regioselective fluorination of, respectively, hydroxyalkyl and oxoalkyl dihydropyran derivatives. PMID- 15787556 TI - Theoretical studies on the mechanism of acid-promoted hydrolysis of N formylaziridine in comparison with formamide. AB - [reaction: see text] We present an ab initio study of the acid-promoted hydrolysis reaction mechanism of N-formylaziridine in comparison with formamide. Since the rate of amide hydrolysis reactions depends on the formation of the tetrahedral intermediate, we focused our attention mainly on the reactant complex, the tetrahedral intermediate, and the transition state connecting these two stationary points. Geometries were optimized using the density functional theory, and the energetics were refined using ab initio theory including electron correlation. Solvent effects were investigated by using polarizable continuum method calculations. The proton-transfer reaction between the O-protonated and N protonated amides was investigated. In acidic media, despite that the N protonated species is more stable than the O-protonated one, it is predicted that both N-protonated and O-protonated pathways compete in the hydrolysis reaction of N-formylaziridine. PMID- 15787557 TI - Synthesis and water solubility of adamantyl-OEG-fullerene hybrids. AB - [reaction: see text] A series of new adamantyl-oligoethyleneglycol-fullerene hybrids was prepared via Bingel-Hirsch functionalization of the C60 fullerene with various adamantyl-oligoethyleneglycol malonates. As NMDA-targeted antioxidants, these compounds may have the potential to be developed as therapeutic agents for the treatment of neurological disorders. PMID- 15787559 TI - Kinetics and mechanism of the aminolysis of O-aryl S-methyl thiocarbonates. AB - [reaction: see text] The reactions of secondary alicyclic (SA) amines and quinuclidines (QUI) with 4-nitrophenyl and 2,4-dinitrophenyl S-methyl thiocarbonates (1 and 2, respectively) and those of SA amines with 2,3,4,5,6 pentafluorophenyl S-methyl thiocarbonate (3) are subjected to a kinetic study in aqueous solution, at 25.0 degrees C, and an ionic strength of 0.2 M (KCl). The reactions of thiocarbonates 1, 2, and 3 were followed spectrophotometrically at 400, 360, and 220 nm, respectively. Under amine excess, pseudo-first-order rate coefficients (k(obsd)) are found. Plots of k(obsd) vs amine concentration at constant pH are linear, with the slope (kN) independent of pH. The Bronsted-type plots (log kN vs pKa of aminium ions) are linear for all the reactions, with slopes beta = 0.9 for those of 1 with SA amines and QUI, beta = 0.36 and 0.57 for the reactions of 2 with SA amines and QUI, respectively, and beta = 0.39 for the reactions of SA amines with 3. The magnitude of the slopes indicates that both aminolyses of 1 are governed by stepwise mechanisms, through a zwitterionic tetrahedral intermediate (T+/-), where expulsion of the nucleofuge from T+/- is the rate-determining step. The values of the Bronsted slopes found for the aminolyses of thiocarbonates 2 and 3 suggest that these reactions are concerted. By comparison of the reactions under investigation between them and with similar aminolyses, the following conclusions arise: (i) Thiocarbonate 2 is more reactive than 1 toward the two amine series. (ii) The change of the nonleaving group from MeO in 4-nitrophenyl methyl carbonate to MeS in thiocarbonate 1 results in lower kN values. (iii) The greater reactivity of this carbonate than thiocarbonate 1 is attributed to steric hindrance of the MeS group, compared to MeO toward amine attack. (iv) The change of a pyridine to an isobasic SA amine or QUI destabilizes the T+/- intermediate formed in the aminolyses of 2. (v) The change of 4 nitrophenoxy to 2,3,4,5,6-pentafluorphenoxy or 2,4-dinitrophenoxy as the leaving group destabilizes the tetrahedral intermediate formed in the reactions with SA amines, changing the mechanism from a stepwise process to a concerted reaction. PMID- 15787558 TI - Regiocontrolled benzannulation of diaryl(gem-dichlorocyclopropyl)methanols for the synthesis of unsymmetrically substituted alpha-arylnaphthalenes: application to total synthesis of natural lignan lactones. AB - [reaction: see text] An efficient synthesis of highly substituted alpha arylnaphthalene analogues has been developed utilizing Lewis acid-promoted regiocontrolled benzannulation of aryl(aryl')-2,2-dichlorocyclopropylmethanols (aryl not equal aryl'; abbreviated as AACMs). Both AACM diastereomers were easily prepared via highly stereoselective addition (>95/5) of ArLi to gem dichlorocyclopropropyl aryl' ketones. The choice of Lewis acids determined the cyclization regioselectivity of the present benzannulation. TiCl4 and SnCl4 used the chelation pathway, whereas silyl triflates used a nonchelation pathway to give unsymmetrically substituted regioisomeric alpha-arylnaphthalenes in 40-91% yields with moderate to excellent regioselectivity (TiCl4 or SnCl4; >99/1-3/1, TBDMSOTf; >1/99-1/4). Thus, the alpha-aryl or alpha-aryl' moiety (accessory aryl group) was alternatively introduced to alpha-arylnaphthalenes by choosing either the order of the reaction sequences or the appropriate catalyst. Application of the present method to the total synthesis for unsymmetrically substituted natural lignan lactones, justicidin B, retrojusticidin B, dehydrodesoxypodophyllotoxin, and a related analogue, 5'-methoxyretrochinensin, was demonstrated. Lignan retrolactones (retrojusticidin B and 5'-methoxyretrochinensin) were synthesized by the conventional lactonization of the diol precursor, whereas a novel Bu2SnO mediated monoacylation method was applied to the synthesis of normal lignan lactones (justicidin B and dehydrodesoxypodophyllotoxin). PMID- 15787560 TI - Cyclopropyl alkynes as mechanistic probes to distinguish between vinyl radical and ionic intermediates. AB - [reaction: see text] The reactions of (trans-2-phenylcyclopropyl)ethyne, 1a, (trans,trans-2-methoxy-3-phenylcyclopropyl)ethyne, 1b, and (trans,trans-2-methoxy 1-methyl-3-phenylcyclopropyl)ethyne, 1c, with either aqueous sulfuric acid or tris(trimethylsilyl)silane (or tributyltin hydride) and AIBN have been investigated. Protonation and addition of the silyl (or stannyl) radical occurred at the terminal position of the alkyne giving an alpha-cyclopropyl-substituted vinyl cation or radical, respectively. Under both reaction conditions, 1a yielded products derived from ring opening toward the phenyl substituent. Alkynes 1b and 1c, however, gave different products depending on whether radical or cationic conditions were used. When radical conditions were employed, products derived from regioselective ring opening toward the phenyl substituent were obtained. In contrast, when cationic conditions were employed, products derived from selective ring opening toward the methoxy substituent were isolated. The corresponding alpha-cyclopropyl-substituted vinyllithium derivatives were also synthesized and were found to be stable toward rearrangement. An estimate of the rate constants for ring opening of the alpha-cyclopropylvinyl cations was also made: values of 10(10)-10(12) s(-1) were found for the vinyl cations derived from protonation of the terminal carbon of alkynes 1a-c. Based on these results, cyclopropyl alkynes 1a-c can be classified as hypersensitive mechanistic probes for the detection of vinyl radical or cationic intermediates generated adjacent to the cyclopropyl ring and, in the case of 1b and 1c, the distinction between a radical or cationic intermediate is possible. PMID- 15787562 TI - Ureidopyrimidinones incorporating a functionalizable p-aminophenyl electron donating group at C-6. AB - [structure: see text] 2-Ureido-4-[1H]-pyrimidinones have been reported to dimerize via quadruple hydrogen bonding systems with dimerization constants >10(6) M(-1) in CDCl3. The dimerization constant, K(dim), is dependent on the solvent as well as the ring-substituents present, where previously alkyl (e.g., R1 = Me) and aromatic moieties (e.g., R1 = p-NO2C6H4, R1 = C6H2(OC13H27)3) have been incorporated at the C-6 position. To assess the influence of alternative, functionalizable, electron-donating groups on the dimerization motif and tautomeric distribution of isomers, the synthesis of compounds possessing aminophenyl functionality at the C-6 position has been achieved. NMR spectroscopy chemical shift analysis revealed that compound 2 (R1 = p-NH2C6H4, R2 = C6H13) existed as the 2-ureido-4-pyrimidinol dimeric DADA array in DMSO-d6, where a dimerization constant of 46 M(-1) was determined. This is the first time that a ureidopyrimidinone quadruple hydrogen bonding DADA array has been observed in pure DMSO, a highly polar solvent. The azo-derivative 5 of compound 2 was prepared which also adopted the pyrimidin-4-ol form in DMSO-d6. Compounds 7, 10 and 11 were then synthesized containing a more hydrophilic PEG unit in the lateral chain and the tautomeric distributions were determined. PMID- 15787561 TI - Synthesis of aryl sulfones via L-proline-promoted CuI-catalyzed coupling reaction of aryl halides with sulfinic acid salts. AB - [reaction: see text] The CuI/L-proline sodium salt catalyzed coupling reaction of aryl halides with sulfinic acid salts readily occurs at 80-95 degrees C in DMSO to give the corresponding aryl sulfones in good to excellent yields. This process is well-tolerated by a wide range of functional groups including hydroxyl, amino, acetanilide, ketone, ester, and nitrile. Using this method, 4-phenylsulfonyl- and 4-methanesulfonyl-substituted L-phenylalanine derivatives are prepared. PMID- 15787563 TI - Dihydrophenanthrene-type intermediates during photoreaction of trans-4'-benzyl-5 styrylfuran. AB - [structure: see text] Photoreaction of trans-4'-benzyl-5-styrylfuran (trans-BSF) has been studied by the 355-nm laser flash photolysis (LFP) in CH2Cl2 using a Nd3+:YAG laser (30 ps, 5 mJ pulse(-1) or 5 ns, 30 mJ pulse(-1)). Transient fluorescence and absorption spectra assigned to the singlet excited trans-BSF were observed during the 30-ps LFP, whereas a transient absorption spectrum with two peaks at 400 and 510 nm, assigned to the trans-fused dihydrophenanthrene (DHP)-type intermediate (DP1), was observed during the 5-ns LFP. It is clearly suggested that a two-photon absorption process is involved in the formation of DP1. The first photoreaction is the photoisomerization of trans-BSF, which occurs to give cis-BSF. The second photoreaction process is photocyclization of cis-BSF, which occurs to give DP1 decaying with the half lifetime (tau1/2) of 2.8-4.0 micros to produce another DHP-type intermediate (DP2) with an absorption peak at 400 nm in the absence of O2, through [1,9]-hydrogen shift. DP2 decayed with tau1/2 > 500 micros to give the product through aromatization. In O2-saturated CH2Cl2, DP1 decayed with tau1/2 = 250 ns to give a radical intermediate (X) with two peaks at 410 and 510 nm, through hydrogen abstraction of DP1 by O2. X decayed with tau1/2 = 150 micros to give the product through successive hydrogen abstraction. PMID- 15787564 TI - Novel carbonyl bromoallylation/Heck reaction sequence. Stereocontrolled access to bicyclic beta-lactams. AB - [reaction: see text] The reaction of 2,3-dibromopropene with racemic as well as enantiopure 4-oxoazetidine-2-carbaldehydes 1 in aqueous media was promoted by tin in the presence of several additives to afford the corresponding bromohomoallyl alcohols 2 in high diastereoselectivities. However, indium or zinc were unable to promote the bromoallylation reaction of aldehydes 1 under similar Barbier-type conditions. Vinyl bromides 2 bearing an extra alkene tether were used for the preparation of differently sized, fused bicyclic beta-lactams of nonconventional structure via Heck cyclization. PMID- 15787565 TI - Generation and reactivity of ketyl radicals with lignin related structures. On the importance of the ketyl pathway in the photoyellowing of lignin containing pulps and papers. AB - [reaction: see text] Ketyl radicals with lignin related structures have been generated by means of radiation chemical and photochemical techniques. In the former studies ketyl radicals are produced by reaction of alpha-carbonyl-beta aryl ether lignin models with the solvated electron produced by pulse radiolysis of an aqueous solution at pH 6.0. The UV-vis spectra of ketyl radicals are characterized by three main absorption bands. The shape and position of these bands slightly change when the spectra are recorded in alkaline solution (pH 11.0) being now assigned to the ketyl radical anions and a pKa = 9.5 is determined for the 1-(3,4,5-trimethoxyphenyl)-2-phenoxyethanol-1-yl radical. Decay rates of ketyl radicals are found to be dose dependent and, at low doses, lie in the range (1.7-2.7) x 10(3) s(-1). In the presence of oxygen a fast decay of the ketyl radicals is observed (k2 = 1.8-2.7 x 10(9) M(-1) s(-1)) that is accompanied by the formation of stable products, i.e., the starting ketones. In the photochemical studies ketyl radicals have been produced by charge-transfer (CT) photoactivation of the electron donor-acceptor salts of methyl viologen (MV2+) with alpha-hydroxy-alpha-phenoxymethyl-aryl acetates. This process leads to the instantaneous formation of the reduced acceptor (methyl viologen radical cation, MV+*), as is clearly shown in a laser flash photolysis experiment by the two absorption bands centered at 390 and 605 nm, and an acyloxyl radical [ArC(CO2*))(OH)CH2(OC6H5)], which undergoes a very fast decarboxylation with formation of the ketyl radicals. Steady-state photoirradiation of the CT ion pairs indicates that 1-aryl-2-phenoxyethanones are formed as primary photoproducts by oxidation of ketyl radicals by MV2+ (under argon) or by molecular oxygen. Small amounts of acetophenones are formed by further photolysis of 1-aryl-2-phenoxyethanones and not by beta-fragmentation of the ketyl radicals. The high reactivity of ketyl radicals with oxygen coupled with the low rates of beta-fragmentation of the same species have an important bearing in the context of the photoyellowing of lignin containing pulps and papers. PMID- 15787566 TI - Improved imide receptors by imprinting using pyrimidine-based fluorescent reporter monomers. AB - [reaction: see text] Optically responsive receptors toward imides based on 6 substituted 2,4-bis(acrylamido)pyrimidines are presented. The monomers were readily prepared in good yield. Solution binding to 1-benzyluracil (BU) monitored by 1H NMR appeared lower than a previously reported pyridine-based monomer. However, as indicated by 1H NMR and IR spectral investigations, the association strength was demonstrated to be "masked" by dimerization of the pyrimidine-based monomer units. Thus, from dilution experiments, a dimerization constant of 731 M( 1) was estimated for the pyrimidine-based monomer 2,4-bis(acrylamido)-6 piperidinopyrimidine whereas for the pyridine-based monomer 2,6 bis(acrylamido)pyridine, no self-association was observed. This precluded an accurate determination of the binding constant for BU to the former monomer whereas for the latter a binding constant of 757 M(-1) was measured. Despite the strong self-association, the novel monomer was shown to lead to enhanced imprinting effects when compared to imprinted polymers prepared analogously, but using the pyridine-based monomer as the recognition element. This was attributed to a higher intrinsic binding affinity exhibited by the pyrimidine based host monomer vis a vis the guest and the existence in the former of more than one interaction site for the guest. The monomers exhibited fluorescence emission informative of the mode of monomer incorporation in the polymer and the presence of guest species. Thus, the fluorescence was rapidly and selectively quenched upon template addition, with the degree of quenching correlating with binding affinity and the amount of template bound to the polymer. PMID- 15787567 TI - Hydrogen-bonded networks in organic conductors: crystal structures and electronic properties of charge-transfer salts of tetracyanoquinodimethane with 4,4' biimidazolium having multiprotonated States. AB - [structure: see text] Novel hydrogen-bonded charge-transfer salts of TCNQ with mono- and diprotonated 4,4'-biimidazolium were synthesized in order to demonstrate the high potential of the 4,4'-biimidazole system in a molecular conductor from the viewpoint of crystal engineering and electronic modulation. Crystal structure analyses of neutral 4,4'-biimidazole and TCNQ salts revealed the formation of two types of hydrogen-bonding modes of the 4,4'-biimidazole moiety depending on the protonated states. Neutral 4,4'-biimidazole possessed a linear chain mode of hydrogen-bonding to construct two-dimensional network. In the TCNQ salt of monoprotonated 4,4'-biimidazolium, the 4,4'-biimidazole moiety formed a dimer by a complementary mode of hydrogen-bonding. In contrast, the salt of diprotonated 4,4'-biimidazolium showed a double linear chain mode of hydrogen bonding to construct a three-dimensional network. The formation of two types of hydrogen-bonding modes made the difference in the stacking patterns of TCNQ columns and in their transport properties. The TCNQ salt of diprotonated 4,4' biimidazolium exhibited high electrical conductivity (sigma(rt) = 1.1 x 10(-1) S cm(-1)). PMID- 15787568 TI - Hexameric and pentameric slipped-cofacial dimers: toward an artificial light harvesting complex. AB - [structure: see text] We prepared a zinc complex of bis(1-methylimidazolyl)-m gable porphyrin. The automatically assembled coordinate species showed a complex mixture of wide molecular weight distributions accompanied with assemblies of specific assembly numbers. When this zinc complex was once dissociated by the addition of methanol and reorganized again by elimination of the methanol under high-dilution conditions in chloroform to facilitate intramolecular coordinate structure formation, two convergent assemblies were obtained through analysis by gel permeation chromatography. These assemblies gave round-shaped particles on solid substrates by probe microscopies (atomic force and scanning tunneling microscopies). These two components were separated by GPC and evidence from small angle X-ray scattering measurements in solution with synchrotron radiation was consistent with hexameric and pentameric macrorings of the gable porphyrin. The porphyrin macrorings did not show any fluorescence quenching by assembly formation, and we anticipate that the macrorings of gable porphyrins represent a good model for an artificial light-harvesting complex. PMID- 15787569 TI - Synthesis and characterization of p-phenylenediamine derivatives bearing an electron-acceptor unit. AB - [reaction: see text] Two series of aniline oligomers bearing fused heterocycles as an electron-acceptor unit were synthesized. They consist of aniline or its derivatives as an electron donor and benzothiadiazole (BT) or quinoxaline (QX) as an electron-acceptor unit. Benzothiadiazoles 1-3 were synthesized by palladium catalyzed amination. Quinoxalines 4-6 were prepared by palladium-catalyzed amination or transformation from the benzothiadiazoles. These compounds showed a HOMO-LUMO gap smaller than those of their analogues such as thiophene-substituted BT/QXs. Cyclic voltammetry revealed that the electrochemical behavior is dependent on the position of the acceptor heterocycle. Chemical oxidation with Ag2O afforded the corresponding 1,4-quinonediimine derivatives as an E,E-isomer, stereoselectively. As for the BT pentamer analogues 2 and 3, the first oxidation selectively occurred at the amino group adjacent to the benzothiadiazole unit, giving the regiospecific half-oxidized derivatives. Furthermore, the fully oxidized derivative 24 was isolated and characterized. PMID- 15787570 TI - Photoenolization of 2-(2-methyl benzoyl) benzoic acid, methyl ester: effect of E photoenol lifetime on the photochemistry. AB - [reaction: see text] Photolysis of 3 in argon-saturated 2-propanol led to formation of 5 via intermolecular H-atom abstraction followed by lactonization. Irradiation of 4 in 2-propanol gave compounds 6 and 7 that also come from intermolecular H-atom abstraction. In contrast, photolysis of an oxygen-saturated solution of 3 in 2-propanol yields products 8, 9, and 10, which were all formed from intramolecular H-atom abstraction and trapping of the corresponding biradical with oxygen. Laser flash photolysis of 3 in methanol showed formation of biradical 3BR (lambda(max) 330 nm, and tau = 50 ns) via intramolecular H-atom abstraction as the main photoreactivity of 3. Biradical 3BR decayed into photoenols 3Z and 3E (lambda(max) 390 nm, tau = 6.5 micros and tau = 162 micros, respectively). In comparison, laser flash photolysis of 4 yielded photoenols 4Z and 4E (lambda(max) 390 nm, tau = 15 micros and tau = 3.6 ms, respectively). Thus photoenol 3E is unusually short-lived, and therefore it does not undergo the intramolecular lactonization as we have observed for the analogous photoenol 1E. Photoenol 3Z decays back to 3 via an intramolecular 1,5-H shift, whereas photoenol 3E reforms 3 efficiently via the solvent with the aid of the ortho ester group. The intramolecular lactonization of photoenols 1E and 3E must be a slow process, presumably because the photoenols are rigid and the hydroxyl group is inhibited, by intramolecular hydrogen bonding, from acquiring the correct geometry for lactonization. Thus only photoenols that are resistant to reformation of their ketone via the solvent are long-lived enough to undergo lactonization and release the alcohol moiety. PMID- 15787571 TI - Synthesis of methyl carbamates from primary aliphatic amines and dimethyl carbonate in supercritical CO2: effects of pressure and cosolvents and chemoselectivity. AB - [reaction: see text] At 130 degrees C, in the presence of CO2 (5-200 bar), primary aliphatic amines react with dimethyl carbonate (MeOCO2Me, DMC) to yield methyl carbamates (RNHCO2Me) and N-methylation side-products (RNHMe and RNMe2). The pressure of CO2 largely influences both the reaction conversion and the selectivity toward urethanes: in general, conversion goes through a maximum (70 80%) in the midrange (40 bar) and drops at lower and higher pressures, whereas selectivity is continuously improved (from 50% up to 90%) by an increase of the pressure. This is explained by the multiple role of CO2 in (i) the acid/base equilibrium with aliphatic amines, (ii) the reactivity/solubility of RNHCO2- nucleophiles with/in DMC, and (iii) the inhibition of competitive N-methylation reaction of the substrates. Cosolvents also affect the reaction: in particular, a drop in selectivity is observed with polar protic media (i.e., MeOH), plausibly because of solvation effects (through H-bonds) of RNHCO2- moieties. The reaction shows also a good chemoselectivity: bifunctional aliphatic amines bearing either aromatic NH2 or OH substituents [XC6H4(CH2)n NH2, X = NH2, OH; n = 1, 2], undergo methoxycarbonylation reactions exclusively at aliphatic amino groups and give the corresponding methyl carbamates [XC6H4(CH2)n NHCO2Me] in 39-65% isolated yields. PMID- 15787572 TI - Triarylethene-based extended pi-systems: programmable synthesis and photophysical properties. AB - [reaction: see text] On the basis of an efficient Pd-catalyzed triarylation to a vinylsilane platform, four types of structurally well-defined triarylethene-based extended pi-systems have been prepared very rapidly. From a compound library constructed by the present method, it was possible to find a number of interesting fluorescent materials, as well as interesting fluorescent properties such as aggregation-induced enhanced emission. A useful method for the rapid synthesis and property evaluation has also been developed. PMID- 15787573 TI - ROM polymerization-capture-release strategy for the chromatography-free synthesis of novel unsymmetrical porphyrazines. AB - [reaction: see text] Crossover-Linstead macrocyclization reactions of two norbornenyl-tagged diaminomaleonitriles with dipropylmaleonitrile gave access to crude mixtures of porphyrazines containing diamino-hexapropyl-porphyrazine magnesium complexes. The mixtures were subjected to ring-opening metathesis polymerization to yield the insoluble diaminoporphyrazine-functionalized polymers. Acid-mediated cleavage from the polymer backbone followed by acylation of the resultant sensitive macrocyclic diamines gave monoacetyl-, monotrifluoroacetyl-, and ditrifluoroacetyl-substituted porphyrazinediamines. Conversion of these amido-porphyrazines to the corresponding zinc macrocycles and studies of their electronic absorption and emission spectra, electrochemistry, and photophysics are described. PMID- 15787574 TI - Buried solvent determines both anion-binding selectivity and binding stoichiometry with hydrogen-bonding receptors. AB - [reaction: see text] The crystal structure of a tetraurea picket porphyrin chloride anion complex has previously shown the anion to be situated between two adjacent ureas and hydrogen bonded via four NH protons (J. Am. Chem. Soc. 1998, 120, 11684-11692). The porphyrin receptor also binds a DMSO molecule and utilizes it as a participant in its anion recognition unit, in a manner similar to enzymes that bind water for use as part of their substrate recognition unit. The bound solvent molecule determines the anion-binding affinity, selectivity, and stoichiometry of binding. With a bound DMSO molecule, the tetraurea picket porphyrin is a highly selective receptor for chloride anion and binds all anions with a 1:1 binding stoichiometry. Absent the buried DMSO molecule, the receptor is selective for phosphate anion and binds chloride and phosphate anions with both 1:1 and 1:2 receptor-anion stoichiometries. Additionally, a remarkable reversal in the selectivity of anion complexation between various picket porphyrin receptors is observed, wherein the binding constant ratios change over 3 orders of magnitude as the receptor's number of urea pickets change from four to two. The latter receptor has no urea pickets available to bind to solvent after complexation with an anion. The results demonstrate that anion complexation with hydrogen-bonding receptors in a competitive solvent is enhanced when a ubiquitous solvent molecule is incorporated into the binding motif. In this way, competitive solvent adds to the overall complexation energy and thereby strengthens binding rather than weakens it, as commonly believed. The results are pertinent to drug design, for they suggest that pharmaceuticals need not be completely desolvated to selectively bind to their biological target when water can be included in the binding motif. PMID- 15787575 TI - Neighboring group participation of 9-anthracenylmethyl group in glycosylation: preparation of unusual C-glycosides. AB - [reaction: see text] A coupling of 2-O-arylmethylated D-glucose-derived thioglycosides with various alcohols in the presence of DMTST as an activator is described. The requisite glycosyl donors are efficiently prepared by one-pot procedures. When the aryl groups are phenyl, p-methoxyphenyl, 1-naphthyl, and 2 naphthyl groups, a mixture of alpha- and beta-anomeric O-glycosides is obtained under the conditions, whereas when the aryl group is the 9-anthracenyl group, a highly stereoselective formation of the unusual C-glycosides in good yields via neighboring group participation of the 9-anthracenylmethyl group followed by coupling with a variety of alcohols is observed. Three new chiral centers including a quaternary carbon are created in one single step. PMID- 15787576 TI - Remarkably mild and simple preparation of sulfenate anions from beta sulfinylesters: a new route to enantioenriched sulfoxides. AB - [reaction: see text] A general, efficient, and experimentally simple method for the generation of sulfenate salts has been developed using beta-sulfinylesters as substrates. The process is based on a retro-Michael reaction, initiated by deprotonation at low temperature. Upon treatment with alkyl halides, the liberated sulfenates are subsequently converted into sulfoxides in good to excellent yield. Extension of the methodology to an unprecedented access to nonracemic sulfoxides by introduction of an enantiopure ligand, (-)-sparteine, is also described. PMID- 15787577 TI - Facile synthesis of oligophenylene-substituted calix[4]arenes and their enhanced binding properties. AB - [reaction: see text] A facile and efficient protocol for the synthesis of oligophenylene OPP(n)-substituted calix[4]arenes (with n up to 4) via iodo substituted oligoarylcalix[4]arenes has been developed. The cooperation effect of the proximate fluoroionophores in hexylsulfanyl end-capped OPP(n)-substituted calix[4]arene assemblies leads to metal ion binding enhancement. PMID- 15787578 TI - Highly substituted tetrahydropyrones from hetero-Diels-Alder reactions of 2 alkenals with stereochemical induction from chiral dienes. AB - [reaction: see text] A new method for the stereoselective synthesis of libraries of 2,3,5-trisubstituted tetrahydro-gamma-pyrones and the corresponding tetrahydropyran-4-ols is reported. Dienes with a chiral moiety at position 5 were synthesized starting from (triphenylphosphoranylidene)acetone. In hetero-Diels Alder (HDA) reactions, especially with alpha,beta-unsaturated aldehydes, they induce diastereomeric ratios from 4:1 to 14:1. Through selective epimerization and reduction, further building blocks are available. These constitute ideal starting points for their use in the total synthesis of complex polyketide macrocycles, especially with the vinyl group available for metathetic coupling. PMID- 15787580 TI - Can the absence of solvation of neutral reagents by ionic liquids be responsible for the high reactivity in base-assisted intramolecular nucleophilic substitutions in these solvents? AB - [reaction: see text] The kinetics of the rearrangement of the Z-phenylhydrazone of 3-benzoyl-5-phenyl-1,2,4-oxadiazole (1a) into the relevant 4-benzoylamino-2,5 diphenyl-1,2,3-triazole (2a) induced by amines have been studied in two room temperature ionic liquids (IL-1, [BMIM][BF4] and IL-2, [BMIM][PF6]). The data collected show that the reaction occurs faster in ionic liquids than in other conventional solvents previously studied (both polar or apolar, protic or aprotic). Presumably, this could depend on their peculiar ability to minimize the strong substrate-solvent, amine-solvent and amine-amine interactions occurring in conventional solvents. PMID- 15787579 TI - Chemoenzymatic asymmetric total synthesis of phosphodiesterase inhibitors: preparation of a polycyclic pyrazolo[3,4-d]pyrimidine from an acylnitroso Diels Alder cycloadduct-derived aminocyclopentenol. AB - [reaction: see text] Enzymatic resolution of Boc-protected 4-aminocyclopenten-1 ol 4c gave both enantiomers 5c and 6c in high ee. Boc removal and separate condensation with chloropyrazolopyrimidine 18 provided elaborated 1,4 aminocyclopentenol derivatives 20 and 26, respectively. Separate treatment of 20 and 26 with Pd(0) under basic conditions induced cyclization to unsaturated polycycles 22 and 27, which, upon catalytic hydrogenation, were transformed to new cyclopentane-containing pyrazolopyrimidines 24 and 28, analogues of recently described novel phosphodiesterase inhibitors. PMID- 15787581 TI - Efficient Stille cross-coupling reaction catalyzed by the Pd(OAc)2/Dabco catalytic system. AB - [reaction: see text] An efficient Pd(OAc)2/Dabco-catalyzed Stille cross-coupling reaction procedure has been developed. In the presence of Pd(OAc)2 and Dabco (triethylenediamine), various aryl halides including aryl iodides, aryl bromides, and activated aryl chlorides were coupled efficiently with organotin compounds to afford the corresponding biaryls, alkene, and alkynes in good to excellent yields. Furthermore, high TONs [turnover numbers, up to 980,000 TONs for the coupling reaction of 1-bromo-4-nitrobenzene and furan-2-yltributyltin] for the Stille cross-coupling reaction were observed. PMID- 15787582 TI - Asymmetric synthesis of bicyclic beta-lactones via the intramolecular, nucleophile-catalyzed aldol lactonization: improved efficiency and expanded scope. AB - [reaction: see text] The intramolecular, nucleophile-catalyzed, aldol lactonization (NCAL) process merges catalytic, asymmetric carbocycle synthesis with beta-lactone synthesis. The application of modified Mukaiyama reagents to this process led to greatly improved conversion and efficiency (70-82% yield) and shorter reaction times with no diminution of enantioselectivity (91-98% ee). The process was extended to several new aldehyde-acid substrates leading to new bicyclic-beta-lactones. This methodology uniquely provides beta-lactone-fused cyclopentanes and cyclohexanes readied for further transformations. PMID- 15787583 TI - Synthesis of 3,4,5-trisubstituted isoxazoles via sequential [3 + 2] cycloaddition/silicon-based cross-coupling reactions. AB - [reaction: see text] A [3 + 2] cycloaddition reaction between alkynyldimethylsilyl ethers and aryl and alkyl nitrile oxides to produce isoxazolylsilanols has been developed. The cross-coupling reactions of these heterocyclic silanols with a variety of aryl iodides affords 3,4,5-trisubstituted isoxazoles. This sequential process allows for rapid variation of substituents at the 3, 4, and 5 positions of the isoxazole. PMID- 15787584 TI - Efficient total synthesis of sapinofuranone B. AB - [structure: see text] An efficient enantioselective synthesis of sapinofuranone B (1) using Sharpless asymmetric dihydroxylation, Sonogashira coupling, and Wittig olefination as the key steps is described. PMID- 15787585 TI - Synthesis of beta-amino-alpha-hydroxy esters and beta-amino-alpha-azido ester by Sharpless asymmetric aminohydroxylation, byproducts analysis. AB - [reaction: see text] Synthesis of enantiomerically pure beta-amino-alpha-hydroxy esters (1, 2) and beta-amino-alpha-azido ester (3) using Sharpless AA as a key step is described. A hitherto unreported side reaction, the oxidation of the beta hydroxy-alpha-amino ester (5) into the alpha,alpha-di-tert-butyloxycarbamoyl-beta ketoester (8) under AA conditions, is documented. PMID- 15787586 TI - Simple preparation of alpha-diazo esters. AB - [reaction: see text] The TiCl4-mediated reaction of an ester with benzoyl chloride results in high yields of the alpha-benzoylated ester. Diazo transfer of the benzoylated ester utilizing p-acetoamidobenzenesulfonyl azide affords the alpha-diazo ester in good yield. Using this simplified procedure, it is easy to prepare gram quantities of alpha-diazo esters. PMID- 15787587 TI - Solvent-free synthesis of functionalized flavones under microwave irradiation. AB - [reaction: see text] Eco-friendly direct solvent-free synthesis of flavones is achieved by microwave irradiation of phloroglucinol and beta-ketoesters. Heating with microwaves versus under classical conditions was shown to be higher yielding, cleaner, and faster. The reaction goes through a cycloaddition of an alpha-oxo ketene intermediate followed by an uncatalyzed thermal Fries rearrangement. PMID- 15787588 TI - Catalytic asymmetric intramolecular aminopalladation: improved palladium(II) catalysts. AB - [reaction: see text] Cobalt oxazoline palladacyclic (COP) complex 4 containing acetate as a bridging ligand is an excellent catalyst for asymmetric intramolecular aminopalladation to synthesize 4-vinyloxazolidin-2-ones in 91-98% ee. In contrast to previously reported Pd(II) catalysts, COP-OAc (4) promotes the asymmetric cyclization of (Z)-allylic N-tosylcarbamates without prior activation by silver salts. PMID- 15787589 TI - Kishner's reduction of 2-furylhydrazone gives 2-methylene-2,3-dihydrofuran, a highly reactive ene in the ene reaction. AB - [reaction: see text] The Kishner reduction of 2-furylhydrazone gives 2-methylene 2,3-dihydrofuran as the major abnormal reduction product. 2-Methylene-2,3 dihydrofuran is an excellent ene in the carbonyl-ene reaction, reacting with a variety of aldehydes. Most notable was the asymmetric carbonyl-ene reaction of 2 methylene-2,3-dihydrofuran and decanal using Ti(OCH(CH3)2)4/(S)-BINOL to give the corresponding alcohol in 66% yield and 94% ee. The reaction of 2-methylene-2,3 dihydrofuran with 2 equiv of 1,4-benzoquinone unexpectedly gave a monoalkylated 1,4-hydroquinone/1,4-benzoquinone electron donor-acceptor complex. PMID- 15787590 TI - Parallel synthesis of strongly fluorescent polysubstituted 2,6-dicyanoanilines via microwave-promoted multicomponent reaction. AB - [reaction: see text] A facile parallel synthesis of polysubstituted 2,6 dicyanoanilines via microwave-promoted three-component reaction of aldehydes, ketones, and propanedinitrile in solution and also on polymer support has been developed. The screening for optical properties identified two new compounds with high fluorescence quantum yields. PMID- 15787591 TI - Polymer-supported organotin reagents for regioselective halogenation of aromatic amines. AB - [reaction: see text] Polymer-supported triorganotin halides were used in the halogenation reaction of aromatic amines. Treatment of aromatic amines with n butyllithium and polymer-supported organotin halides gave the corresponding polymer-bound N-triorganostannylamines, which by treatment with bromine or iodine monochloride gave the para-halogenated aromatic amines with high yields and high selectivities. The polymer-supported organotin halides reagents regenerated during the course of the halogenation reaction can be reused without loss of efficiency. The presence of tin residues in halogenated aromatic amines was also investigated and evaluated at under 20 ppm after three runs. PMID- 15787592 TI - Homogeneous supported synthesis using ionic liquid supports: tunable separation properties. AB - [structure: see text] We report a homogeneous supported version of Koser's salt based on a room-temperature ionic liquid (RTIL) support. By altering the nature of the RTIL, a material was developed that was stable, recyclable, and readily separable from the tosyloxylated ketone products just by using variations in solvent polarity. A similar approach should be applicable to a wide range of supported catalysts and reagents. PMID- 15787593 TI - Regioselective one-pot synthesis of 9-alkyl-6-chloropyrido[3,2 e][1,2,4]triazolo[4,3-a]pyrazines. Reactivity of aliphatic and aromatic hydrazides. AB - [reaction: see text] The one-pot synthesis of new 9-alkyl-6-chloropyrido[3,2 e][1,2,4]triazolo[4,3-a]pyrazines has been achieved. Hydrazides regioselectively reacted as nucleophiles with the 3-chloro substituent of 2,3-dichloropyrido[2,3 b]pyrazine. An intramolecular cyclization afforded the tricycle nonxanthine adenosine receptor antagonists. PMID- 15787594 TI - Trifluoroacetic acid-mediated hydroarylation: synthesis of dihydrocoumarins and dihydroquinolones. AB - [reaction: see text] Trifluoroacetic acid mediates the hydroarylation of alkenes to afford dihydrocoumarins and dihydroquinolones in good yield. Intermolecular hydroarylation of cinnamic acids by phenols is particularly facile, which leads to the conclusion that previous reports of palladium-catalyzed hydroarylation of cinnamic acids in trifluoroacetic acid are erroneous. PMID- 15787598 TI - Molecular mousetraps and the serpinopathies. AB - Members of the serine proteinase inhibitor or serpin superfamily inhibit their target proteinases by a remarkable conformational transition that involves the enzyme being translocated more than 70 A (1 A = 10(-10) m) from the upper to the lower pole of the inhibitor. This elegant mechanism is subverted by point mutations to form ordered polymers that are retained within the endoplasmic reticulum of secretory cells. The accumulation of polymers underlies the retention of mutants of alpha(1)-antitrypsin and neuroserpin within hepatocytes and neurons to cause cirrhosis and dementia respectively. The formation of polymers results in the failure to secrete mutants of other members of the serpin superfamily: antithrombin, C1 inhibitor and alpha1-antichymotrypsin, to cause a plasma deficiency that results in the clinical syndromes of thrombosis, angio oedema and emphysema respectively. Understanding the common mechanism underlying the retention and deficiency of mutants of the serpins has allowed us to group these conditions as the serpinopathies. We review in this paper the molecular and structural basis of the serpinopathies and show how this has allowed the development of specific agents to block the polymerization that underlies disease. PMID- 15787599 TI - A genomic view of the complexity of mammalian proteolytic systems. AB - Proteolytic enzymes play an essential role in different physiological processes, including development, reproduction and host defence, as well as in numerous pathologies, like inflammatory diseases, neurological disorders or cancer. The completion of the human genome sequence allowed us to determine that more than 2% of all human genes are proteases or protease inhibitors, reflecting the importance of proteolysis in human biology. To understand better the complexity of proteases in human and other model organisms, we have used the available genome sequences of different mammalian organisms, including mouse, rat and chimpanzee, to identify and compare their degradomes, the complete set of protease genes in these species. Surprisingly, the rodent protease complement is more complex when compared with that of primates, mainly due to the expansion of protease families implicated in reproduction and host defence. Similarly, most differences between human and chimpanzee proteases are found in genes implicated in the immune system, which might explain some of the differences between both organisms. We have also found several genes implicated in reproduction, nutrition and the immune system, which are functional in rat, mouse or chimpanzee, but have been inactivated by mutations in the human lineage. These findings suggest that pseudogenization of specific protease genes has been a mechanism contributing to the evolution of the human genome. Finally, we found that proteases implicated in human hereditary diseases, and especially in neurodegenerative disorders, are highly conserved among mammals. PMID- 15787600 TI - Roles of proteolysis and lipid rafts in the processing of the amyloid precursor protein and prion protein. AB - In the amyloidogenic pathway, the APP (amyloid precursor protein) is proteolytically processed by the beta- and gamma-secretases to release the Abeta (amyloid-beta) peptide that is neurotoxic and aggregates in the brains of patients suffering from Alzheimer's disease. In the non-amyloidogenic pathway, APP is cleaved by alpha-secretase within the Abeta domain, precluding deposition of intact Abeta peptide. The cellular form of the PrP(C) (prion protein) undergoes reactive oxygen species-mediated beta-cleavage within the copper binding octapeptide repeats or, alternatively, alpha-cleavage within the central hydrophobic neurotoxic domain. In addition, PrP(C) is shed from the membrane by the action of a zinc metalloprotease. Members of the ADAM (a disintegrin and metalloproteinase) family of zinc metalloproteases, notably ADAM10 and TACE (ADAM17) display alpha-secretase activity towards APP and appear to be responsible for the alpha-cleavage of PrP(C). The amyloidogenic cleavage of APP by the beta- and gamma-secretases appears to occur preferentially in cholesterol rich lipid rafts, while the conversion of PrP(C) into the infectious form PrP(Sc) also appears to occur in these membrane domains. PMID- 15787601 TI - Introduction to the Kinases in Diabetes Biochemical Society Focused Meeting: are protein kinases good targets for antidiabetic drugs? AB - Insulin regulates whole-body glucose homoeostasis by modulating the activities of protein kinases in its target tissues: muscle, liver and fat. Defects in insulin's ability to modulate protein kinase activity lead to 'insulin resistance' or impaired insulin action. Insulin resistance in combination with defective insulin secretion from the pancreas results in the elevated blood glucose levels that are characteristic of diabetes mellitus. Pharmacological agents that selectively modulate protein kinase activities in insulin-resistant tissues may act either as insulin-sensitizing or insulin-mimetic drugs. Consistent with this, small molecule modulators of a number of protein kinases have demonstrated efficacy in animal models of insulin resistance and diabetes. Moreover, emerging data in humans suggest that marketed anti-diabetic agents may also act in part through modulating protein kinase activities. This meeting was convened to consider the potential to treat insulin resistance and Type II diabetes by modulating protein kinase activity. PMID- 15787602 TI - Therapeutic challenges of kinase and phosphatase inhibition and use in anti diabetic strategy. AB - The development of kinase and phosphatase inhibitors as novel therapeutic agents has been stimulated by the discovery that most biological processes are controlled by the reversible phosphorylation of proteins. Most of the early results in this area were generated in oncology, at the same time as the human genome, with its 500+ kinases and 100+ phosphatases was deciphered. Because of this, we know a great deal about which processes signalling inhibitors interfere with, but little about the overall consequences. In this study, kinases will be briefly reviewed, followed by some of the early problems in developing kinase inhibitors, as biochemical reagents, and clinically active pharmaceuticals in oncology. The discussion will then switch to the potential role of kinases and phosphatases in controlling the disease process in Type II diabetes. Phosphatase inhibitors should augment insulin receptor tyrosine kinase signalling. Glycogen synthesis and glycogenolysis are phosphorylation dependent, and amenable to kinase inhibition, as are some nuclear hormone receptors, and these will be briefly discussed. PMID- 15787603 TI - Role of protein kinase B in insulin-regulated glucose uptake. AB - The activation of protein kinase B (or Akt) plays a central role in the stimulation of glucose uptake by insulin. Currently, however, numerous questions remain unanswered regarding the role of this kinase in bringing about this effect. For example, we do not know precisely where in the GLUT4 trafficking pathway this kinase acts. Nor do we know which protein substrates are responsible for mediating the effects of protein kinase B, although two recently identified proteins (AS160 and PIKfyve) may play a role. This paper addresses these important questions by reviewing recent progress in the field. PMID- 15787604 TI - Atypical protein kinase C in insulin action and insulin resistance. AB - It now seems clear that aPKC (atypical protein kinase C) isoforms are required for insulin-stimulated glucose transport in muscle and adipocytes. Moreover, there are marked defects in the activation of aPKCs under a variety of insulin resistant conditions in humans, monkeys and rodents. In humans, defects in aPKC in muscle are seen in Type II diabetes and its precursors, obesity, the obesity associated polycystic ovary syndrome and impaired glucose tolerance. These defects in muscle aPKC activation are due to both impaired activation of insulin receptor substrate-1-dependent PI3K (phosphoinositide 3-kinase) and the direct activation of aPKCs by the lipid product of PI3K, PI-3,4,5-(PO4)3. Although it is still uncertain which underlying defect comes first, the resultant defect in aPKC activation in muscle most certainly contributes significantly to the development of skeletal muscle insulin resistance. Of further note, unlike the seemingly ubiquitous presence of defective aPKC activation in skeletal muscle in insulin resistant states, the activation of aPKC is normal or increased in livers of Type II diabetic and obese rodents. The maintenance of aPKC activation in the liver may explain how insulin-dependent lipid synthesis is maintained in these states, as aPKCs function mainly in the activation of enzymes important for lipid synthesis. Thus increased activation of liver aPKC in hyperinsulinaemic states may contribute significantly to the development of hyperlipidaemia in insulin resistant states. PMID- 15787605 TI - Insulin signal transduction in human skeletal muscle: identifying the defects in Type II diabetes. AB - Type II diabetes is characterized by defects in insulin action on peripheral tissues, such as skeletal muscle, adipose tissue and liver and pancreatic beta cell defects. Since the skeletal muscle accounts for approx. 75% of whole body insulin-stimulated glucose uptake, defects in this tissue play a major role in the impaired glucose homoeostasis in Type II diabetic patients. Thus identifying defective steps in this process may reveal attractive targets for drug development to combat insulin resistance and Type II diabetes. This review will describe the effects of insulin on glucose transport and other metabolic events in skeletal muscle that are mediated by intracellular signalling cascades. Evidence for impaired activation of the insulin receptor signalling cascade and defective glucose transporter 4 translocation in the skeletal muscle from Type II diabetic patients will be presented. Through the identification of the intracellular defects in insulin action that control glucose homoeostasis, a better understanding of the disease pathogenesis can be gained and strategies for intervention may be developed. PMID- 15787606 TI - Modulation of insulin signalling by insulin sensitizers. AB - Insulin resistance is a hallmark of Type II diabetes. It is well documented that insulin sensitizers such as peroxisome-proliferator-activated receptor gamma agonists and aspirin improve insulin action in vivo. The detailed mechanisms by which the insulin sensitizers promote insulin signalling, however, are not completely understood and remain somewhat controversial. In the present review, we summarize our studies attempting to explore the molecular mechanisms underlying the effects of insulin sensitizers in cells and in animal models of insulin resistance. In 3T3-L1 adipocytes and/or in HEK-293 cells stably expressing recombinant IRS1 protein (insulin receptor substrate protein 1), the peroxisome-proliferator-activated receptor gamma agonist rosiglitazone and aspirin promote insulin signalling by decreasing inhibitory IRS1 serine phosphorylation. Increased IRS1 Ser-307 phosphorylation and concomitant decreased insulin signalling as measured by insulin-stimulated IRS1 tyrosine phosphorylation and Akt threonine phosphorylation were observed in adipose tissues of Zucker obese rats compared with lean control rats. Treatment with rosiglitazone for 24 and 48 h increased insulin signalling and decreased IRS1 Ser 307 phosphorylation concomitantly. Treatment of the Zucker obese rats with rosiglitazone for 24 h also reversed the high circulating levels of free fatty acids, which have been shown to correlate with increased IRS1 serine phosphorylation. Taken together, the results suggest that IRS1 inhibitory serine phosphorylation is a key component of insulin resistance and its reversal may be physiologically relevant to insulin sensitization in vivo. PMID- 15787607 TI - AMP-activated protein kinase and the metabolic syndrome. AB - The occurrence of Type II (non-insulin-dependent) diabetes and obesity and their associated morbidities continue to increase and they are rapidly reaching epidemic proportions. AMPK (AMP-activated protein kinase) was initially thought of as an intracellular 'fuel gauge' responding to a decrease in the level of ATP by increasing energy production and decreasing energy utilization. Recent studies have shown that AMPK plays a role in controlling the whole body energy homoeostasis, including the regulation of plasma glucose levels, fatty acid oxidation and glycogen metabolism. In addition to its effects on the periphery, AMPK has been found to play a key role in the control of food intake through its regulation by hormones, including leptin, within the hypothalamus. The control of AMPK activity, therefore, provides an attractive target for therapeutic intervention in metabolic disorders such as obesity and Type II diabetes. Indeed, a number of physiological and pharmacological factors that are beneficial in these disorders have been shown to act, at least in part, through the activation of AMPK. PMID- 15787608 TI - PDH kinase inhibitors: a novel therapy for Type II diabetes? AB - The pyruvate dehydrogenase multienzyme complex catalyses the oxidative decarboxylation of pyruvate, which is an important regulatory step in oxidative metabolism. Phosphorylation of the E1 (pyruvate decarboxylase) subunit on one of three specific serine residues results in loss of enzyme activity. Four dedicated PDHK (pyruvate dehydrogenase kinase) isoenzymes have been identified, each of which display a distinct tissue-specific expression profile, and have differential regulatory properties. Thus PDHK play a key role in controlling the balance between glucose and lipid oxidation according to substrate supply. Increasing glucose oxidation by inhibiting PDHK may be an effective mechanism to increase glucose utilization; additionally, increasing pyruvate oxidation may further contribute to lowering of glucose level by decreasing the supply of gluconeogenic substrates. A number of PDHK inhibitors are now available to enable this mechanism to be evaluated as a therapy for diabetes. The isoenzyme selectivity profile of AZD7545 and related compounds will be described and evidence for their non-ATP-competitive mode of action presented. These compounds increase PDH activity in vivo, and when dosed chronically, improve glycaemic control in Zucker rats. Furthermore, glucose lowering has been demonstrated in the hyperglycaemic Zucker diabetic fatty rat. This result supports the hypothesis that inhibition of PDHK may be an effective therapy for Type II diabetes. PMID- 15787609 TI - Small molecule glucokinase activators as novel anti-diabetic agents. AB - The monomeric enzyme GK (glucokinase) has a low affinity for glucose and, quantitatively, is largely expressed in the liver and pancreatic beta-cells, playing a key 'glucose sensing' role to regulate hepatic glucose balance and insulin secretion. Mutations of GK in man can be inactivating, to cause a form of diabetes mellitus, or activating, to lower blood glucose levels. Recently, models of GK protein structure have helped to elucidate the role of inactivating and activating mutations, with the latter revealing an allosteric binding site, possibly for an unknown physiological activator. However, this discovery was pre dated by Drug Discovery projects that have identified small organic molecules that activate pancreatic and liver GK enzyme activity. These compounds stimulate insulin secretion in islets and glucose metabolism in hepatocytes. The profile of these GK activators, both in vitro and in vivo and the potential role that GK activators play in lowering blood glucose levels in Type II diabetes mellitus will be discussed. PMID- 15787610 TI - Before and beyond ABA: upstream sensing and internal signals that determine ABA accumulation and response under abiotic stress. AB - Sensing and signalling events that detect abiotic stress-induced changes in plant water status and initiate downstream stress responses such as ABA (abscisic acid) accumulation and osmoregulation remain uncharacterized in plants. Although conclusive results are lacking, recent results from plants, and analogies to signalling in other organisms, suggest possible mechanisms for sensing altered water status and initial transduction of that signal. Internal signals that act downstream of ABA and modulate stress responses to reflect the type and severity of the stress and the metabolic status of the plant are also not well understood. Two specific types of signalling, sugar sensing and reactive oxygen signalling, are likely to be modulators of ABA response under stress. For both upstream sensing and signalling of plant water status as well as downstream modulation of ABA response, present results suggest several genetic strategies with high potential to increase our understanding of the molecular basis by which plants sense and respond to altered water status. PMID- 15787611 TI - Storage reserve mobilization in germinating oilseeds: Arabidopsis as a model system. AB - Germinating oilseeds break down fatty acids through peroxisomal beta-oxidation and convert the carbon into soluble carbohydrates through the glyoxylate cycle and gluconeogenesis. This interconversion is unique among higher eukaryotes. Using a combination of forward and reverse genetic screens, we have isolated mutants that compromise fatty acid breakdown at each step. These mutants exhibit characteristic, yet nonidentical, seedling establishment phenotypes that can be rescued by the provision of an alternative carbon source. In addition, we have recently shown that Arabidopsis seed's lipid breakdown occurs in two distinct tissues, the embryo and endosperm. The utilization of endospermic lipid reserves requires gluconeogenesis and transport of the resulting sugars to the germinating embryo. We discuss the potential of the Arabidopsis endosperm tissue as a simplified model system for the study of germination and lipid breakdown in germinating oilseeds. PMID- 15787612 TI - The molecular biology of appressorium turgor generation by the rice blast fungus Magnaporthe grisea. AB - The rice blast fungus Magnaporthe grisea develops specialized infection structures known as appressoria, which develop enormous turgor pressure to bring about plant infection. Turgor is generated by accumulation of compatible solutes, including glycerol, which is synthesized in large quantities in the appressorium. Glycogen, trehalose and lipids represent the most abundant storage products in M. grisea conidia. Trehalose and glycogen are rapidly degraded during conidial germination and it is known that trehalose synthesis is required for virulence of the fungus. Lipid bodies are transported to the developing appressoria and degraded at the onset of turgor generation, in a process that is cAMP-dependent. A combined biochemical and genetic approach is being used to dissect the process of turgor generation in the rice blast fungus. PMID- 15787613 TI - Serpentine plant MLO proteins as entry portals for powdery mildew fungi. AB - In the dicotyledonous plant species Arabidopsis and the monocot barley, presence of specific isoforms of the family of heptahelical plasma membrane-localized MLO proteins is required for successful host-cell invasion by ascomycete powdery mildew fungi. Absence of these MLO proteins, either caused by natural polymorphisms or induced lesions in the respective Mlo genes, results in failure of fungal sporelings to penetrate the plant cell wall. As a consequence, recessively inherited cell-autonomous mlo resistance is effective against all known isolates of powdery mildew fungi colonizing either barley or Arabidopsis. Barley MLO interacts constitutively with the cytoplasmic calcium sensor calmodulin, but the strength of this interaction increases transiently during fungal pathogenesis. In addition, MLO as well as ROR2, a plasma membrane-resident syntaxin also implicated in mlo penetration resistance, focally accumulate at sites of attempted fungal attack, thereby defining a novel pathogen-triggered micro-domain. In conclusion, powdery mildew fungi appear to specifically corrupt MLO to modulate vesicle-associated processes at the plant cell periphery for successful pathogenesis. PMID- 15787614 TI - Post-translational regulation in plants employing a diverse set of polypeptide tags. AB - The concept that plants exploit polypeptides as post-translational modifiers is rapidly emerging as an important method to manipulate various cellular processes. The best known is Ub (ubiquitin) that serves as reusable tag for selective protein degradation by the 26 S proteasome and for endosomal trafficking. Genomic analyses indicate that Ub pathway alone comprises over 6% of the Arabidopsis proteome with thousands of proteins being targets. Consequently, this pathway influences much of plant biology. Others tags include RUB-1 (related to Ub-1; also known as NEDD8), SUMO (small Ub-like modifier), ATG-8 (autophagy-8) and ATG 12, UFM-1 (Ub-fold modifier-1) and HUB-1 (homology to Ub-1). Preliminary studies indicate that these tags have much more limited sets of targets and provide more specialized functions, including transcriptional regulation, protein localization, autophagic turnover and antagonizing the effects of Ub. On the basis of their widespread distribution and pervasive functions, peptide tags can now be considered as prime players in plant cell regulation. PMID- 15787615 TI - Coagulation inhibitors in inflammation. AB - Coagulation is triggered by inflammatory mediators in a number of ways. However, to prevent unwanted clot formation, several natural anticoagulant mechanisms exist, such as the antithrombin-heparin mechanism, the tissue factor pathway inhibitor mechanism and the protein C anticoagulant pathway. This review examines the ways in which these pathways are down-regulated by inflammation, thus limiting clot formation and decreasing the natural anti-inflammatory mechanisms that these pathways possess. PMID- 15787616 TI - Soluble adhesion molecules in inflammatory and vascular diseases. AB - For many years the vascular endothelium was believed simply to provide a passive lining between circulating blood and extravascular tissue. It is now clear, however, that this monolayer of cells on the luminal surface of all blood vessels, provides a selective barrier that responds dynamically to various stimuli, and controls a complex series of cellular reactions and interactions. The current presentation describes the use of computer enhanced video recording to study interactions between endothelial cells and circulating blood cells, especially leucocytes. Subsequently, modern assays for soluble cell adhesion molecules and other cell receptors were assessed for potential use in routine clinical practice. The results demonstrated that adhesive mechanisms involving leucocytes and endothelial cells involve a range of interrelationships that cut across conventional views of haemostasis and leucocyte function. The findings also suggest that interplay between the vascular lumen and circulating blood cells might be vitally important in clinically demanding pathologies, such as life-threatening sepsis, ischaemic heart disease, atherosclerosis and cancer. The concepts provide challenging strategies for further investigation. PMID- 15787617 TI - Using proteomics to identify potential therapeutic targets in platelets. AB - Proteomics has provided powerful new insights into the complex events of the anucleate platelet and has revealed many potential protein targets in the search for suitable agents for thrombotic disease. In the present study, we summarize recent proteomic approaches to analyse specific platelet subproteomes, such as the platelet releasate, the platelet phosphotyrosine proteome and characterization of the proteins associated with membrane lipid rafts. PMID- 15787618 TI - Harnessing wound healing and regeneration for tissue engineering. AB - Biomedical science has made major advances in understanding how cells grow into functioning tissue and the signalling mechanisms used to achieve this are slowly being dissected. Tissue engineering is the application of that knowledge to the building or repairing of organs, including skin, the largest organ in the body. Generally, engineered tissue is a combination of living cells and a supporting matrix. Besides serving as burn coverings, engineered skin substitutes can help patients with diabetic foot ulcers. Today, most of these ulcers are treated with an approach that includes antibiotics, glucose control, special shoes and frequent cleaning and bandaging. The results of such treatments are often disappointing and ineffectual, and scarring remains a major problem, mechanically, cosmetically and psychologically. Within our group we are attempting to address this by investigating novel approaches to skin tissue engineering. We are identifying novel therapeutic manipulations to improve the degree of integration between a tissue engineered dermal construct and the host by both molecular manipulation of growth factors but also by understanding and harnessing mechanisms of regenerative biology. For the purpose of this summary, we will concentrate primarily on the latter of these two approaches in that we have identified a novel mouse mutant that completely and perfectly regenerates skin and cartilaginous components following ear injury. This experimental animal will allow us to characterize not only novel genes involved in the regeneration process but also to utilize cells from such animals in artificial skin equivalents to assess their behaviour compared with normal cells. This approach should allow us to create a tissue-engineered substitute, which more closely resembles the normal regional microanatomy and physiology of the skin, allowing better integration to the host with minimal or no scarring. PMID- 15787619 TI - The role of blood cells and their microparticles in blood coagulation. AB - The transmembrane glycoprotein TF (tissue factor) plays an essential role in haemostasis as the principal initiator of blood coagulation. In this paper, we describe how the circulating blood cells--monocytes, platelets, neutrophils and their microparticles--co-operate in regulating the expression, availability and activity of monocyte-derived TF. PMID- 15787620 TI - Polyunsaturated fatty acids and inflammation. AB - The n-6 polyunsaturated fatty acid, arachidonic acid, is a precursor of prostaglandins, leukotrienes and related compounds that have important roles as mediators and regulators of inflammation. Consuming increased amounts of long chain n-3 polyunsaturated fatty acids (found in oily fish and fish oils) results in a partial replacement of the arachidonic acid in cell membranes by eicosapentaenoic and docosahexaenoic acids. This leads to decreased production of arachidonic acid-derived mediators. This alone is a potentially beneficial anti inflammatory effect of n-3 fatty acids. However, n-3 fatty acids have a number of other effects that might occur downstream of altered eicosanoid production or are independent of this. For example, they result in suppressed production of pro inflammatory cytokines and can modulate adhesion molecule expression. These effects occur at the level of altered gene expression. PMID- 15787621 TI - Haemostasis in normal pregnancy: a balancing act? AB - Pregnancy is a risk factor for venous thrombosis and the incidence of venous thromboembolism during normal pregnancy is 6-fold higher during pregnancy than in the general female population of child-bearing age. This incidence is, however, remarkably low given the increases in markers of haemostatic activation observed during normal pregnancy. During normal healthy pregnancy there are substantial changes in the haemostatic system, many of which are procoagulant and supposed to be in preparation for the haemostatic challenge of delivery. Normal haemostasis requires a balance between coagulation and fibrinolysis to maintain the integrity of the vasculature, and complex physiological changes are evident during pregnancy which appear to ensure a constant coagulation/fibrinolysis balance. This balance is maintained, at least partly, by an increase in fibrinolytic activity, but decreases in other factors such as factor XI and monocyte tissue factor expression may also serve to counterbalance procoagulant changes. PMID- 15787622 TI - What about the rest of the patients? PMID- 15787623 TI - Platelet activation markers in patients with heart assist device. AB - Clinical use of heart assist devices is often associated with thromboembolic complications. We hypothesized that platelets may be activated in patients receiving assist devices and examined expression of the platelet activation markers CD62, CD63, and thrombospondin using flow cytometry in eight patients with Novacor left ventricular assist system (LVAS) or Berlin Heart. Patients with end-stage heart failure had elevated expression of platelet activation markers before insertion of the assist device. While CD62 (P < 0.05) and thrombospondin expression (n.s.) decreased by the 14th postoperative day, the CD63 expression remained elevated (n.s.). A good correlation was found between CD62 and thrombospondin expression (r = 0.72). Bleeding time ex vivo indicated platelet dysfunction during the first 4 weeks after implantation. No relation between expression of platelet activation markers and bleeding time ex vivo were found. In conclusion, expression of the platelet activation markers CD62, CD63, and thrombospondin is increased in patients with end-stage heart failure before device placement and shows prolonged elevation during the assist period. Future studies in larger patient populations are necessary to identify new and specific markers of platelet activation in this clinical setting. PMID- 15787624 TI - Descending thoracic aortic aneurysm repair with the aid of partial cardiopulmonary bypass: heparin-coated circuits versus nonheparin-coated circuits. AB - BACKGROUND: We have performed descending thoracic aortic aneurysm repairs with partial cardiopulmonary bypass, including heparin-coated circuits. The aim of this study was to evaluate (i) the impact of partial cardiopulmonary bypass on distal organ function and surgical outcomes; and (ii) the effectiveness of using heparin-coated circuits for preventing bleeding complications. METHODS: From July 1980 to June 2004, 309 patients underwent descending thoracic aortic aneurysm repairs using partial cardiopulmonary bypass. Their mean age was 61 years (range 19-81 years). One hundred of the 309 patients underwent repair of descending thoracic aortic aneurysm with heparin-coated circuits. Blood data for renal and hepatic function were collected on the day before the operation and postoperative days. RESULTS: The in-hospital mortality was 15%. Distal organ dysfunction included spinal cord dysfunction in 2 patients (0.7%) and renal failure necessitating hemodialysis in 15 patients (5%, 15/297: excluded 12 dialysis patients). Multivariate analyses showed that preoperative hemodialysis and emergency operation were risk factors for operative mortality and that emergency operation was a risk factor for requiring hemodialysis. Renal and hepatic function normalized by 2 weeks after surgery. There were no significant differences between the heparin-coated group and nonheparin-coated group in amounts of packed red cells, fresh frozen plasma, and platelets transfused during the procedures. CONCLUSIONS: Our data showed that partial cardiopulmonary bypass is a safe and effective method for distal perfusion. Using this technique, descending thoracic aortic aneurysm repair can be performed with acceptable mortality and morbidity. However, the superiority of heparin-coated circuits over nonheparin-coated ones was not proved. PMID- 15787625 TI - Impact of inverse plasma filtration on complement activation in porcine blood. AB - During extracorporeal circulation, activation of the complement system as a bioincompatibility reaction upon contact with artificial surfaces is well known. This may induce adverse effects for the patient as well as for the bioreactor when using a hybrid liver support system. At present, the achievable clearance rates in hybrid liver support systems are restricted by the limitations of blood flow through the capillaries of the plasma filter. In order to enhance clearance rates it is desirable to develop a high-flow plasma separation unit in which the activation of the complement system is as low as possible. Conventional plasma filters operating in normal and in inverse (blood flow around the capillaries and the capillary lumen serving as plasma compartment) modes were investigated applying an in vitro circuit (n = 4 in each group). For this purpose heparinized porcine blood was used. Complement activation was measured as generation of the soluble membrane attack complex, SC5b-9. With a normal flow mode the maximum achievable filtration rates varied from 25 to 40 mL/min, whereas the maximal filtration rates with an inverse flow mode varied from 40 to 100 mL/min. While these values were significantly higher than those with regularly operating plasma filters, there was no significant difference between normal and inverse mode in the production of SC5b-9. We conclude that the outer surface of the investigated conventional plasma filtration membranes is suitable for an inverse mode and does not lead to an excessive activation of the complement system upon dynamic blood contact. Thus, the inversion of the blood and the plasma compartments may be a possible way forward in the development of a high-flow plasma separation filter. PMID- 15787626 TI - Protein L-agarose for adsorption of autoantibodies: a potential tool for extracorporeal treatment. AB - This work investigated the adsorption of autoantibodies such as anti-SS-A/Ro, anti-SS-B/La, anti-Sm, and anti-dsDNA on protein L-agarose gel. In order to determine better conditions for IgG adsorption on this matrix, some buffer systems were tested. Adsorption data were analyzed using the Langmuir and Langmuir-Freundlich isotherm models. The experimental isotherms were best described by the Langmuir-Freundlich model, which indicated negative and positive cooperativities for binding in the presence of PBS and HEPES buffers, respectively. The K(d) values for phosphate buffered saline solution (PBS) and hydroxyethylpiperazine ethanesulfonic acid (HEPES) were 2.8 x 10(-7) M and 3.2 x 10(-7) M, respectively, which indicate a high affinity between IgG and the immobilized protein L. The amount of protein adsorbed per amount of protein loaded was high for anti-Sm (44%) and anti-dsDNA (46%), but low for anti-SS-B/La (9%). The amount of albumin adsorbed was lower than 0.06 mg/mL, which may remove the need for a plasma replacement solution in clinical apheresis. PMID- 15787628 TI - Thyroid disorders in hemodialysis patients in an iodine-deficient community. AB - There are various changes in the thyroid gland and its function in chronic renal failure (CRF). These changes include lower levels of circulating thyroid hormone, altered peripheral hormone metabolism, decreased binding to carrier proteins, possible reduction in tissue hormone content, and increased iodine storage in the thyroid gland. The decrease of excretion of urinary iodine in CRF increases serum inorganic iodine level and iodine content of the thyroid, which consequently enlarges the gland. This study is designed to investigate the prevalence of goiter and thyroid dysfunction in patients with end-stage renal disease (ESRD) on hemodialysis (HD) in an iodine-deficient community. Eighty-seven (40 females and 47 males) HD patients and 169 (79 females and 90 males) healthy individuals as controls are included. Sex ratios for the patient and control groups are 0.85 and 0.88, respectively. Mean ages for the patient and control groups are 42.94 +/- 11.88 and 40.20 +/- 10.72 years, respectively. Examination of the thyroid gland using ultrasonography along with simultaneous measurement of blood levels of free T4 (FT4), free-T3 (FT3), and thyrotropin (TSH) are made for every individual. The presence of goiter demonstrable by ultrasonography is found in 32.2% of the uremic patients and in 23.5% of the controls and its prevalence increases with age (P = 0.01). In 32 (36.8%) of the patients and 29 (17.1%) of the controls at least one thyroid nodule is found in ultrasonography. Between patients with or without a nodular goiter the authors could not observe any difference for duration of dialysis and serum levels of TSH, FT4, FT3, calcium, and albumin. In ESRD patients the prevalence of nodular goiter is higher for females (47.5% vs. 27.7%, P = 0.045) and increases with age (P = 0.04). Though incidence of hyperthyroidism is found to be similar for the two groups (1.14% in ESRD patients vs. 1.10% in controls), hypothyroidism is observed in 3.4% of ESRD patients but only 0.6% of controls. This high incidence of hypothyroidism and nodular goiter in ESRD patients shows that screening for thyroid dysfunction and goiter, using appropriate laboratory tests and ultrasonography, should be considered in evaluation of every ESRD patient. PMID- 15787627 TI - Effectiveness of combining plasma exchange and continuous hemodiafiltration in patients with postoperative liver failure. AB - Nine patients with postoperative liver failure were treated with plasma exchange (PE) or PE and continuous hemodiafiltration (CHDF), and various biochemical parameters were determined before and after treatment. Although citrate levels increased significantly after treatment compared with pretreatment levels in both the PE group and the PE + CHDF group (P < 0.0001 and P < 0.0001, respectively), the percentage of the increase in citrate levels was significantly higher in the PE group than in the PE + CHDF group (P = 0.0051). Total bilirubin (T-Bil) levels were significantly lower after treatment in both the PE and PE + CHDF groups (P < 0.0001 and P = 0.0001, respectively). There were no significant differences in T Bil levels between the two groups (P = 0.5181). There were no significant differences in interleukin (IL)-6 levels before and after treatment in both the PE and PE + CHDF groups (P = 0.1281 and P = 0.2273, respectively). IL-18 levels were significantly lower after treatment in both the PE and PE + CHDF groups (P < 0.0001 and P = 0.0002, respectively), but there were no significant differences in the removal rate of IL-18 in both the PE and PE + CHDF groups (P = 0.8749). These results indicate that combining PE and CHDF in a series-parallel circuit is an effective modality for suppressing the elevation of blood citrate levels. This finding may have important implications for the development of an effective treatment for patients with postoperative liver failure. PMID- 15787629 TI - Renal-portal shunt ameliorates renovascular hypertension in pigs. AB - BACKGROUND: Renovascular hypertension is the most common curable form of secondary hypertension. Renin angiotensin system activation depends on the balance between renin production by the kidney and renin degradation by the liver. Thus, we aimed to examine whether deviation of renin-rich blood from the affected kidney into the portal circulation (portalization) can ameliorate renovascular hypertension. METHODS: We selected a porcine model of unilateral renal artery stenosis because the pig's anatomy and physiology are comparable to those of humans and because pigs have already been found capable of developing chronic renovascular hypertension. Angiography and ultrasonography were deliberately used to evaluate renal artery stenosis and the renal-portal shunt. Histology was used to examine the effects of portalization on the kidney and liver after a period of two months. RESULTS: As expected, following the creation of a left renal artery stenosis both renin activity and mean blood pressure measurements increased from 1.23 +/- 0.06 ng/mL/h and 85.6 +/- 0.5 mm Hg at baseline to 4.59 +/- 0.02 ng/mL/h and 126 +/- 1.76 mm Hg, respectively. After portalization renin activity returned to the normal range (1.59 +/- 0.07 ng/mL/h) followed by a concomitant reduction of mean blood pressure to 91 +/- 2 mm Hg. Moreover, a significant correlation was observed between changes in renin activity and blood pressure measurements during the two stages of the experiment. Both the kidney and liver remained macroscopically and microscopically intact at the end of the experiment. CONCLUSION: Portalization of the affected kidney can ameliorate renovascular hypertension and therefore, it might be of benefit in those individuals with fibromascular or atheromatous lesions in the renal artery or its branches not amenable to balloon angioplasty or surgical revascularization. PMID- 15787630 TI - Influence of growth hormone on maintenance of capillary-like structures in an in vitro model of stromal vascular tissue--results from morphometric analysis. AB - The in vitro development of a vascular stroma might be a solution for the engineering of vascularized tissues, however, in vitro stability of capillary like structures is limited. In order to test the influence on maintenance of capillary-like structures, human growth hormone (hGH) was added in concentrations of 0.5, 5, 50, and 500 ng/mL in an in vitro model of stromal vascular tissue. The angiogenic response and maintenance of capillary-like structures were analyzed by means of confocal laser scanning microscopy (CLSM) and image analysis after 8, 16, and 32 days of culture. The highest angiogenic response was observed with a concentration of 50 ng/mL hGH. With the addition of 50 and 500 ng/mL, the length of capillary-like structures could be maintained on high levels up to the 32nd day of culture, whereas with 5 ng/mL values dropped to the level of the control group. The proposed technique of analysis allows quantification of capillary-like network formation and might be useful for tissue engineering applications. PMID- 15787631 TI - Design and initial testing of a mock human circulatory loop for left ventricular assist device performance testing. AB - A mock circulatory loop, which simulates the human circulatory system, is needed to bench test the various versions of continuous flow (CF) left ventricular assist devices (LVADs). This article describes the design and initial testing of such a loop. The loop consists of: (1) pulsatile left and right cardiac simulators; (2) air/water tanks to model the venous and arterial compliances; (3) tygon tubes to model the venous, arterial, and other system flow resistances; and (4) a tuning clamp to model the variation in system resistance characteristics under different cardiac pressure/flow conditions. Several loop measurements were carried out without an LVAD to verify the cardiovascular modeling of a healthy person in sleep, rest, and physical activity, and in different pathological states, and compared to the data found in the literature to validate the loop performance prior to LVAD testing. PMID- 15787632 TI - The hemolytic characteristics of monopivot magnetic suspension blood pumps with washout holes. AB - The hemolytic characteristics of monopivot magnetic suspension blood pumps as a function of impeller washout hole configuration and female pivot shape are observed. The pump impellers are designed with three washout hole configurations for blood circulation, and four female pivot shapes to reduce blood stagnation and to enhance antithrombogenicity. The hemolytic characteristics of the monopivot pumps were observed to be better than those of a currently available commercial centrifugal blood pump, BP-80, and changed to be nearly equal when the female pivot shape was changed. This indicates that hemolysis in monopivot pumps is mainly caused by shear stress between the male and female pivots. PMID- 15787633 TI - Fluorescence bronchoscopy for selection of surgical procedure in patients with early staged endobronchial carcinoma. AB - BACKGROUND: It is sometimes difficult to determine the extent of resection in patients with endobronchial carcinoma because preoperative white-light bronchoscopic (WLB) examination is not sensitive enough to examine the extent fully. Light-Induced Fluorescence Endoscopy (LIFE) is recognized as a useful modality for the diagnosis of early staged bronchial carcinoma, but there have been no reports of its significance in surgical treatment. We have studied the influence of LIFE upon the selection of surgical procedures or other treatments in patients with endobronchial carcinoma preoperatively. METHODS: Conventional WLB and LIFE were performed within 7 days of operation. Biopsy specimens were taken from the marginal regions of the areas that were suspicious for malignancy on LIFE examination and WLB. We decided the resection line before operation and kept the resected central margin 1 cm apart from the area revealed as suspicious by LIFE. RESULTS: From January 1999 to March 2003, 75 patients underwent LIFE. LIFE was performed to decide the surgical procedures for 12 patients (16.0%). Surgical therapy was performed in 8 (66.7%). LIFE findings dramatically changed the surgical procedures for 3 patients, lobectomy changed to sleeve lobectomy in 2 and laser therapy to segmentectomy in 1. LIFE revealed larger abnormal areas of bronchial cancer compared to WLB in 7 (58.3%). Conversely, WLB overestimated the extent of abnormal area in 1 (8.3%). WLB revealed the same cancer area as LIFE in 4. In all patients, the resected bronchial margins were cancer free intraoperatively and postoperatively. CONCLUSION: LIFE can be more sensitive than WLB and be more beneficial for judging the extent of neoplastic bronchial changes for some patients. LIFE may be a useful modality for the preoperative selection of surgical procedures, especially whether sleeve resection is needed or not, for some centrally located superficial endobronchial carcinoma. PMID- 15787634 TI - Correlation between serum nafamostat mesilate and activated coagulation time during continuous hemodiafiltration. AB - We analyzed the relationship between nafamostat mesilate (NM) level and activated coagulation time (ACT) as measured using an Actalyke MINI system vs. an ACTester system in patients with fulminant hepatitis undergoing continuous hemodiafiltration. The hemofilter was made with a polyacrylonitrile membrane, and NM was used as the anticoagulant. Significant correlation was found between the NM dose and the prehemofilter ACT measured by the Actalyke MINI system (r = 0.561, P = 0.0352, n = 14). However, no significant correlation was found between the NM dose and the prehemofilter ACT measured by the ACTester. Neither was a significant correlation found between the prehemofilter NM level and the prehemofilter ACT measured by either system. No significant correlation was found between the NM dose and the posthemofilter ACT measured by either system. The ACTester uses a test tube containing celite and a small amount of kaolin as a coagulation activator. The results of the present study suggest that the ACTester can adsorb NM and that this feature can affect ACT measurement. Considering this influence and the need to minimize the dose of NM in patients with fulminant hepatitis to avoid hemorrhagic tendencies, it is advisable to use the Actalyke MINI system. PMID- 15787638 TI - Cloning and sequencing of Indian water buffalo interleukin-18 cDNA. AB - Summary Full-length cDNA (582 bp) of the interleukin-18 (IL-18) gene of the Indian water buffalo (Bubalus bubalis) was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) and sequenced. The deduced amino acid sequence has 99% and 95% similarity with the IL-18 sequences of cattle and sheep, respectively. There are two amino acid substitutions at positions 132 and 182 in buffalo IL-18 compared with that of cattle. Phylogenetic analysis showed that the IL-18 sequence of fish forms a different lineage and is most divergent from that of cattle, buffalo, sheep, pig, dog, horse, human, monkey, mouse, rat and chicken. PMID- 15787639 TI - Novel variations in the signal peptide region of transforming growth factor beta1 gene in patients with hepatitis: a brief report from India. AB - Genotypic status of the signal peptide region of transforming growth factor beta1 (TGF-beta1) showed a significant difference in C/C-genotype frequency at +29 position (codon 10) between a range of viral hepatitis patients and controls (P = 0.009, OR = 3.15, CI = 1.29-7.678), contributed by those who were infected with hepatitis B virus (HBV) alone or HBV + hepatitis delta virus (HDV) (P = 0.003, OR = 5.0, CI = 1.78-13.97). PMID- 15787640 TI - A comparison of methods for determining genotypes at the tumour necrosis factor alpha-308, interleukin (IL)-1beta+3953, IL-6 -174 and IL-10 -1082/-819/-592 polymorphic loci. AB - Induced heteroduplex genotyping (IHG) is one of many methods that can be used to determine single nucleotide polymorphisms (SNPs). It is relatively new in comparison to other polymerase chain reaction (PCR)-based techniques. The aim of this study was to compare the results of genotyping using IHG with the results of genotyping using either polymerase chain reaction-sequence-specific primers (PCR SSP) or polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) for SNPs in the tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6 and IL-10 genes. Ninety patients who consented to participate in the study had their genotypes determined by IHG and either PCR-SSP (TNF-alpha-308 and IL-10 -1082/-819/-592) or PCR-RFLP (IL-1beta +3953 and IL-6 -174). Results for each locus were compared between techniques by calculating the Kappa statistic as a measure of agreement. The IHG and more traditional genotyping methods produced very similar results at all loci. The Kappa statistics for each locus were as follows: TNF-alpha -308, K = 0.727; IL-1beta +3953, K = 0.886; IL-6 -174, K = 0.909; IL-10 -1082, K = 0.876; IL-10 -592, K = 0.920. IHG is a valid method for the determination of genotypes at the loci examined in this study and produces comparable results to those of more traditional methods of genotyping. PMID- 15787641 TI - Polymorphisms of the upstream regulatory region of the major histocompatibility complex DRB genes in domestic horses. AB - Sequence information was obtained on the variation of the ELA-DRB upstream regulatory region (URR) after polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) cloning and sequencing of approximately 220 bp upstream of the first exon of horse DRB genes. The sequence of the proximal URR of equine DRB is composed of highly conserved sequence motifs, showing the presence of the W, X, Y, CAAT and TATA conserved boxes of major histocompatibility complex (MHC) class II promoters. Five different polymorphic horse DRB promoter sequences were detected in five horse breeds. The results demonstrate the existence of polymorphism in the nucleotide sequences of the ELA DRB URR, located in the functionally important conserved consensus sequences, the X2 box, the Y box and the TATA box, while conservation were observed in X1 and CAAT boxes. The nucleotide diversity among horse URRs was intermediate between that seen within human and mouse DRB promoters, suggesting the existence of another important source of variability in ELA-DRB genes. In addition, phylogenetic comparisons, identity analysis and sequence organization suggested that the reported sequences would correspond to an expressed ELA-DRB locus. However, further information about the functional significance of these promoter polymorphisms will probably be acquired through expression studies on the different sequences. PMID- 15787642 TI - Distribution of human chemokine (C-X3-C) receptor 1 (CX3CR1) gene polymorphisms and haplotypes of the CC chemokine receptor 5 (CCR5) promoter in Chinese people, and the effects of CCR5 haplotypes on CCR5 expression. AB - Two chemokine (C-X3-C) receptor 1 (CX3CR1) gene polymorphisms, V249I and T280M, and 10 CC chemokine receptor 5 (CCR5) promoter haplotypes, P1-P10, have recently been reported to influence the progression of acquired immune-deficiency syndrome (AIDS). As these studies were performed mainly with Caucasian and African American subjects, we determined the distribution of these alleles in Chinese people for the purpose of predicting possible clinical responses to the human immunodeficiency virus type 1 (HIV) epidemics in countries with significant Chinese populations, as well as to establish their effects on the expression of surface CCR5. Ninety-six HIV-negative Chinese individuals in Taiwan were subjected to genotyping, and we thus determined that the allelic frequencies of CX3CR1V249I and T280M changes were 2.6% and 2.1%, respectively, which were lower than found in Caucasians (25.5% and 14.0%, respectively). Unlike the previous reports, we only detected CCR5P1 and P4 haplotypes in Taiwanese people, and the P1/P1, P1/P4 and P4/P4 genotype frequencies were 21.0%, 41.1% and 37.9%, respectively. The sequencing data confirmed the results of previous studies, showing that CCR5P1 exhibited a complete linkage disequilibrium with a polymorphic allele 59029A present in the CCR5 promoter. Furthermore, fluorescence activated cell sorter analysis revealed that, in the absence of the CCR2-64I mutation, individuals carrying CCR5P1 tended to express more surface CCR5 on monocytes and CD4+ cells. Therefore, this study not only reports the frequencies for the CX3CR1 and CCR5 promoter haplotypes in a Chinese population living in Taiwan, but also identifies a statistical link between the P1/P1 haplotype and the elevated CCR5 expression levels in the study group. PMID- 15787643 TI - Nomenclature for factors of the HLA system, 2004. PMID- 15787644 TI - Nomenclature for factors of the HLA system, update November 2004. PMID- 15787645 TI - Nomenclature for factors of the HLA system, update December 2004. PMID- 15787646 TI - Fine needle aspiration cytology in the diagnosis of bone lesions. AB - OBJECTIVE: Fine needle aspiration cytology (FNAC) in combination with radiological examination has recently gained clinical recognition for evaluating skeletal lesions. We evaluated our experience with the use of FNA in diagnosing bone lesions with emphasis on areas of difficulty and limitations. MATERIALS AND METHODS: Over a period of 5 years FNA was performed in 66 cases of bone lesions. Aspirations were done by cytopathologists using 22-gauge needle. Out of 66 cases unsatisfactory aspirate was obtained in 12 cases. Cytohistological correlation was available in 19 cases. RESULTS: Adequate aspirates were categorized into neoplastic (27 cases) and non-neoplastic (27 cases) lesions. Of the 27 neoplastic aspirates, 20 were malignant (12 primary, 8 metastatic deposits) and 7 were benign. In the malignant group osteosarcoma was correctly diagnosed in 3 cases while other 3 were labeled as sarcoma NOS because of lack of osteoid. Metastatic deposits were sub-typed in 6 cases; from renal cell carcinoma (3 cases), proststic adenocarcinoma, follicular carcinoma thyroid, and squamous cell carcinoma. Neoplastic group comprised of 6 cases of cysts and 21 cases of chronic osteomyelitis. Thirteen cases were diagnosed as tuberculous osteomyelitis. CONCLUSIONS: FNA is a frequent indication in metastases in the bone where distinct cytologic features can even identify an unknown primary. However, diagnosis of primary tumours of the bone is limited by precise subtyping of the tumours. FNA has emerged as a cost effective tool for initial diagnosis of both neoplastic and non-neoplastic lesions of the bone. PMID- 15787647 TI - Pleomorphic adenoma of salivary gland: to what extent does fine needle aspiration cytology reflect histopathological features? AB - OBJECTIVE: The histological diversity encountered in pleomorphic adenoma may cause diagnostic difficulty in fine needle aspiration (FNA) cytology due to limited and selective sampling. The present study based on 25 histologically confirmed pleomorphic adenoma cases attempts to find out to what extent FNA cytology reflects the histopathological features. METHODS: May-Grunwald-Giemsa and Papanicolaou stained smears, and haematoxylin and eosin stained paraffin sections of 25 pleomorphic adenomas of parotid and submandibular glands were reviewed. The cellularity, which was assessed in a sliding scale of 1+ to 4+, and proportions of epithelial to mesenchymal components in FNA smears and histology was determined and compared. The frequency of morphological features such as squamous metaplastic cells, cells with oncocytic change, acinus formation, mucus globules, papilla formation, giant cells, myxoid and chondroid matrix as well as specific nuclear features was compared between the two diagnostic methods, and the statistical significance was determined using Fisher's exact test of probability. RESULTS: There was complete concordance between cytology and histology with respect to overall cellularity in 14 (56.0%) cases and in the proportions of epithelial to mesenchymal components in 13 (52.0%). Epithelial cells and myxoid matrix were present in all cases. There was no significant difference between smear and tissue section with respect to frequency of squamous metaplasia, oncocytic change, acinus formation, papilla formation, mucus globules, giant cells, nuclear pleomorphism, nuclear chromatin pattern, and mitotic figures. Morphological parameters that were significantly higher in FNAC compared with histology included intranuclear cytoplasmic inclusions (36.0% versus 8.0%, p = 0.0374), nuclear grooves (84.0% versus 48.0%, p = 0.0090), and reniform nuclei (20.0% versus 0.0%, p = 0.0502). Chondroid matrix was the only parameter which was significantly more common in histology than in cytology (44.0% versus 4.0%, p = 0.019). CONCLUSION: FNA cytology demonstrates well most of the histological features of pleomorphic adenoma of salivary gland and may be considered a useful tool in initial assessment of the tumour. PMID- 15787648 TI - The reliability and accuracy of intraoperative imprint cytology of sentinel lymph nodes in breast cancer. AB - OBJECTIVE: Sentinel lymph node (SLN) biopsy is a new component of the surgical treatment of breast cancer that accurately predicts axillary status. In this study the authors evaluated the accuracy of intraoperative imprint cytology (IC) in comparison with definitive histologic evaluation of SLN in breast cancer patients. METHODS: A total 413 women with breast carcinoma and clinically negative axillary nodes underwent breast surgery and SLN biopsy. Mapping of SLN involved injection of (99m)Technecium labelled human albumin nanocolloid particles and Patent Blue dye. At the Department of Pathology, SLNs were bisected along its major axis. Both halves were imprinted 2-4 times on the slides and immediate staining with Hemacolor (Merck Germany) was performed for intraoperative examination. Imprint node negative women underwent no further surgery, while node positive women proceeded to full axillary clearance. Histological analysis of the SLN involved serial sectioning of the whole node with H&E and immunostaining for cytokeratin. RESULTS: Definitive histology revealed metastases (pN+) in 159/413 patients (38.5%): 69 (16.7%) macro metastases, 57 (13.8%) micro metastases, and 33 (8%) women with only isolated IHC positive cells or positive cell groups smaller than 0.2 mm (pNO sn+). The other 254 women had negative SLN biopsy. Imprint cytology detected 54/69 macro metastases, and 4/57 micro metastases. In the group with negative SLN (254), 2 cases were ''false positives''. CONCLUSIONS: Imprint of SLN biopsy can identify a negative axilla with high accuracy (specificity 99.2%). Overall sensitivity is only 36.5%, but macrometastases are detected in 77% which is important for performing ALDN in one session with operation of primary tumour. PMID- 15787649 TI - Laboratory performance measures: evidence against low-risk women explaining low detection rates of high-grade abnormalities. AB - OBJECTIVE: When a laboratory has a low reporting rate of high-grade abnormality for its cervical cytology specimens, the question arises whether this is due to the laboratory screening a low-risk group of women. This study was undertaken to explore the hypothesis that a low-risk group of women were screened by Victorian laboratories not meeting the recommended minimum standard for the detection of high-grade abnormalities. METHODS: A cohort of 28 094 women was established comprising women whose cervical cytology was reported in 2000 by laboratories not meeting the recommended standard of 0.5% high-grade abnormalities in their reporting of community smears. Outcome measures included the prevalence of high grade abnormality on the next cytology test for the women, the positive predictive value of the cytology reports of high-grade abnormality, and the standardized incidence ratio for a later diagnosis of cervical cancer. RESULTS: The prevalence of high-grade abnormality was 0.36% on the index cytology compared with 0.68% on the first subsequent cytology report. Sixty-nine per cent (60/87) of the index cytology reports of high-grade abnormality were confirmed as high grade abnormalities on histology, compared with 70.8% (121/171) for the first subsequent reports of high-grade abnormality. During 70 015 person-years at risk, nine cases of cervical cancer were observed compared with 5.21 expected cases, giving a standardized incidence ratio of 1.73 (95% CI 0.79-3.28). CONCLUSIONS: These results do not support a hypothesis that the low detection rate for high grade abnormalities is due to the women comprising a low-risk group for cervical neoplasia. PMID- 15787650 TI - Comparison of DNA extraction from cervical cells collected in PreservCyt solution for the amplification of Chlamydia trachomatis. AB - OBJECTIVE: The aim of this study was to compare and evaluate three methods of DNA extraction for the amplification of Chlamydia trachomatis in uterine cervical samples collected in PreservCyt solution. ThinPrep is the trade name for the slide preparation. METHODS: Thirty-eight samples collected in LCx buffer medium, which were identified as C. trachomatis infected by ligase chain reaction (LCR), were selected for this study. DNA from the PreservCyt samples was extracted by three methods: (i) QIAamp kit, (ii) boiling in Tris-EDTA buffer with Chelex purification, and (iii) Proteinase K digestion with Chelex purification. Sample DNA was tested for the presence of C. trachomatis by PCR using cryptic plasmid research (CTP) primers and major outer membrane protein research momp gene (MOMP) primers. Real-time (LightCycler) PCR for relative C. trachomatis quantification following DNA extraction was performed using primers (Hsp 60) for the 60 kDa heat shock protein hsp60 gene. RESULTS: Amplification using CTP primers was the most successful with each of the extraction protocols. Boiling in buffer was the least successful extraction method. QIAamp was the best extraction method, yielding the most positives with both the CTP and MOMP primers. Proteinase K-Chelex extraction gave similar sensitivity to QIAamp extraction with CTP primers but lower for MOMP primers. CONCLUSIONS: The DNA extraction method must be carefully selected to ensure that larger PCR amplicons can be successfully produced by PCR and to ensure high sensitivity of detection of C. trachomatis. In this study it was found that the QIAamp extraction method followed by PCR with the CTP primers was the most successful for amplification of C. trachomatis DNA. PMID- 15787651 TI - P-cadherin expression in glandular lesions of the uterine cervix detected by liquid-based cytology. AB - OBJECTIVE: To study P-cadherin aberrant expression as a possible marker for cervical adenocarcinomas in cytological samples. METHODS: We studied P-cadherin immunoexpression in liquid-based cervical cytology samples of biopsy-proven cervical lesions. RESULTS: We found a statistically significant correlation between P-cadherin expression and a cytological diagnosis of malignancy, either glandular or squamous (P < 0.0001). Twenty-two of 33 malignant cases showed P cadherin membrane staining. None of the 30 benign cases tested showed membrane staining, but three of them displayed an aberrant nuclear P-cadherin expression. CONCLUSIONS: We concluded that P-cadherin can be used to discriminate between malignant and benign cervical cytological specimens, but not to discriminate glandular from squamous lesions. PMID- 15787652 TI - Cytomorphological spectrum in scar endometriosis: a study of eight cases. AB - OBJECTIVE: Endometriosis due to ectopic endometrial tissue that responds to hormonal stimulation and is extremely rare in a surgical scar appearing in 0.1% of women who have undergone Caesarean section. Fine needle aspiration cytology (FNAC) can be a valuable diagnostic aid in the evaluation of these subcutaneous abdominal masses. METHODS: We present the cytomorphological spectrum in eight cases of abdominal wall endometriosis diagnosed by FNAC over a 6-year period (June 1998-June 2004) in Mubarak Al-Kabeer Hospital (Kuwait). The patients ranged from 27 to 56 years of age. Seven had a prior Caesarean section and one had a hysterectomy for fibroid. They presented 3-8 years later with nodules in/near the abdominal scar. Five cases presented with a painful nodule, fluctuant with the menstrual cycle. RESULTS: Cytological findings comprised epithelial clusters and fusiform stromal cells with numerous haemosiderin-laden macrophages. Mild to moderate epithelial atypia was observed in three cases. A diagnosis of endometriosis was rendered in all the eight cases and tissue was available in four cases. CONCLUSIONS: Endometriotic nodules need to be differentiated from other benign/malignant masses and evaluated for possible malignant transformation. FNAC provides a safe and effective tool for diagnosis thereby obviating the need for other procedures. PMID- 15787653 TI - Cytopathology in Belgium. PMID- 15787654 TI - A parotid mass with unusual cytological features. PMID- 15787655 TI - Response to 'Direct booking colposcopy clinic -- the Portsmouth experience'. PMID- 15787657 TI - Animal models of diabetes mellitus. AB - Animal models have been used extensively in diabetes research. Early studies used pancreatectomised dogs to confirm the central role of the pancreas in glucose homeostasis, culminating in the discovery and purification of insulin. Today, animal experimentation is contentious and subject to legal and ethical restrictions that vary throughout the world. Most experiments are carried out on rodents, although some studies are still performed on larger animals. Several toxins, including streptozotocin and alloxan, induce hyperglycaemia in rats and mice. Selective inbreeding has produced several strains of animal that are considered reasonable models of Type 1 diabetes, Type 2 diabetes and related phenotypes such as obesity and insulin resistance. Apart from their use in studying the pathogenesis of the disease and its complications, all new treatments for diabetes, including islet cell transplantation and preventative strategies, are initially investigated in animals. In recent years, molecular biological techniques have produced a large number of new animal models for the study of diabetes, including knock-in, generalized knock-out and tissue-specific knockout mice. PMID- 15787658 TI - How to ensure your paper is rejected by the statistical reviewer. PMID- 15787659 TI - Twice-daily pre-mixed insulin rather than basal insulin therapy alone results in better overall glycaemic control in patients with Type 2 diabetes. AB - AIMS: To compare the glycaemic control of an insulin lispro mixture (25% insulin lispro and 75% NPL) twice daily in combination with metformin to that of once daily insulin glargine plus metformin in patients with Type 2 diabetes inadequately controlled with intermediate insulin, or insulin plus oral agent(s) combination therapy. RESEARCH DESIGN AND METHODS: Ninety-seven patients were randomized in a multicentre, open-label, 32-week crossover study. Primary variables evaluated: haemoglobin A1c (A1c), 2-h post-prandial blood glucose (BG), hypoglycaemia rate (episodes/patient/30 days), incidence (% patients experiencing > or = 1 episode) of overall and nocturnal hypoglycaemia. RESULTS: At endpoint, A1c was lower with the insulin lispro mixture plus metformin compared with glargine plus metformin (7.54% +/- 0.87% vs. 8.14% +/- 1.03%, P < 0.001). Change in A1c from baseline to endpoint was greater with the insulin lispro mixture plus metformin (-1.00% vs. -0.42%; P < 0.001). Two-hour post-prandial BG was lower after morning, midday, and evening meals (P < 0.001) during treatment with the insulin lispro mixture plus metformin. The fasting BG values were lower with glargine plus metformin (P = 0.007). Despite lower BG at 03.00 hours (P < 0.01), patients treated with the insulin lispro mixture plus metformin had a lower rate of nocturnal hypoglycaemia (0.14 +/- 0.49 vs. 0.34 +/- 0.85 episodes/patient/30 days; P = 0.002), although the overall hypoglycaemia rate was not different between treatments (0.61 +/- 1.41 vs. 0.44 +/- 1.07 episodes/patient/30 days; P = 0.477). CONCLUSION: In patients with Type 2 diabetes and inadequate glucose control while on insulin or insulin and oral agent(s) combination therapy, treatment with a twice-daily insulin lispro mixture plus metformin, which targets both post-prandial and pre-meal BG, provided clinically significant improvements in A1c, significantly reduced post-prandial BG after each meal, and reduced nocturnal hypoglycaemia as compared with once-daily glargine plus metformin, a treatment that targets fasting BG. PMID- 15787660 TI - Optimization of basal insulin delivery in Type 1 diabetes: a retrospective study on the use of continuous subcutaneous insulin infusion and insulin glargine. AB - AIMS: To compare the effects on glycaemic control after using continuous subcutaneous insulin infusion (CSII) or insulin glargine. METHODS: Data were obtained from 17 diabetes outpatient clinics in Sweden, employing the same diabetes data management system. Type 1 diabetic patients using multiple dose injections were included prior to starting on either CSII (n = 563) or glargine (n = 513). The median duration of therapy was 25 months for CSII and 6 months for glargine. The comparison between the treatment modalities was carried out by multiple regression analysis and logistic regression analysis in an attempt at reducing the influence of confounding factors. RESULTS: The mean HbA1c decrease was 0.59 +/- 1.19% for CSII and 0.20 +/- 1.07% for glargine (P < 0.001, when assessed by logistic regression). An additional 0.1% lower HbA1c would be expected if glargine had been optimized with basal insulin 40-60% of the daily dose. The more pronounced effect of CSII was achieved with a lower daily dosage of insulin. In a multiple regression analysis with a change of HbA1c as the dependent variable, the following variables were significant: choice of treatment (P < 0.001), HbA1c prior to treatment (P < 0.001) and BMI prior to treatment (P < 0.01). CONCLUSION: Both regimes improved metabolic control, but CSII resulted in significantly higher reduction in HbA1c than after insulin glargine treatment, particularly in those individuals who had higher levels of HbA1c at baseline. PMID- 15787661 TI - Tumour necrosis factor receptors (TNFRs) in Type 2 diabetes. Analysis of soluble plasma fractions and genetic variations of TNFR2 gene in a case-control study. AB - AIMS: We have studied the relationships between soluble fractions of tumour necrosis factor receptors (sTNFR1 and sTNFR2) in Type 2 diabetes (DM2) and its chronic microvascular complications. Likewise, we have analysed the genetic susceptibility of 196T > G exon6/CA-repeat intron 4 mutations in the TNFR2 gene in this population. METHODS: A case-control study was conducted to examine the role of sTNFRs in 345 DM2 patients and 173 healthy subjects. The mutations were studied in all healthy subjects and in a subset of 232 patients. RESULTS: sTNFRs levels were similar in healthy and DM2 patients. A positive correlation between age and both sTNFRs was observed in healthy subjects. In DM2 patients, sTNFR1 showed a positive correlation with age, systolic blood pressure and leptin levels (r = 0.53, P < 0.0001; r = 0.28, P = 0.005; r = 0.46, P < 0.0001, respectively) and sTNFR2 was positively correlated with age, triglycerides and leptin levels (r = 0.34, P < 0.0001; r = 0.21, P < 0.0001; r = 0.28, P = 0.002, respectively). Patients with micro- or macroalbuminuria showed higher plasma levels of sTNFR1 and sTNFR2 than normoalbuminuric patients, after adjusting for confounding variables (B = 0.85, P = 0.022, 95% CI: 0.12-1.58 for sTNFR1 and B = 1.50, P < 0.001, 95% CI: 0.67-2.33 for sTNFR2). In DM2 patients, TT-exon 6 homozygous showed lower levels of sTNFR1 [2,4 (1.1) vs. 3.4 (1.2) ng/ml], and the CA273 allele tracked with elevated plasma HDL-cholesterol [1.8 (0.7), 1.4 (0.3) and 1.3 (0.3) mm, for CA273/273, CA273/- and CA-/-, respectively]. No association was seen with other analysed variables. CONCLUSIONS: Our findings suggest that chronic TNF activation may have some pathogenic role in diabetic nephropathy in DM2 patients. Genetic variations in exon 6/intron 4 of the TNFR2 gene do not predispose to a major risk for DM2 or its microvascular complications. PMID- 15787662 TI - Electrical spinal cord stimulation in the long-term treatment of chronic painful diabetic neuropathy. AB - AIMS: Electrical spinal cord stimulation (ESCS) is a technique for the management of chronic painful diabetic neuropathy (CPDN) affecting the lower limbs. We assessed the efficacy and complication rate of ESCS implanted at least 7 years previously in eight patients. METHODS: After a trial period of percutaneous stimulation, eight male patients had been implanted with a permanent system. Mean age at implantation was 53.5 years and all patients were insulin treated with stage 3 severe disabling CPDN of at least 1 year's duration. The ESCS was removed from one patient at 4 months because of system failure and one patient died 2 months after implantation from a myocardial infarction. RESULTS: Six patients were reviewed a mean of 3.3 years post-implantation. With the stimulator off, McGill pain questionnaire (MPQ) scores (a measure of the quality and severity of pain) were similar to MPQ scores prior to ESCS insertion. Pain scores (visual analogue scale) were measured with the stimulator off and on, respectively: background pain [74.5 (63-79) mm vs. 25 (17-33) mm, median (interquartile range), P = 0.03), peak pain (85 (80-92) mm vs. 19 (11-47) mm, P = 0.03]. There were two further cardiovascular deaths (these patients had continued pain relief) and the four surviving patients were reassessed at 7.5 (range 7-8.5) years: background pain [73 (65-77) mm vs. 33 (28-36) mm, median (interquartile range)], peak pain [86 (81-94) mm vs. 42 (31-53) mm]. Late complications (> 6 months post-insertion) occurred in two patients; electrode damage secondary to trauma requiring replacement (n = 1), and skin peeling under the transmitter site (n = 1). One patient had a second electrode implanted in the cervical region which relieved typical neuropathic hand pains. CONCLUSIONS: ESCS can continue to provide significant pain relief over a prolonged period of time with little associated morbidity. PMID- 15787663 TI - A long-term comparison of pioglitazone and gliclazide in patients with Type 2 diabetes mellitus: a randomized, double-blind, parallel-group comparison trial. AB - AIMS: This study compared the effects of pioglitazone and gliclazide on metabolic control in drug-naive patients with Type 2 diabetes mellitus. METHODS: A total of 1270 patients with Type 2 diabetes were randomized in a parallel-group, double dummy, double-blind study. Patients with poorly controlled Type 2 diabetes (HbA1c 7.5-11%), despite dietary advice, received either pioglitazone up to 45 mg once daily or gliclazide up to 160 mg two times daily. Primary efficacy endpoint was change in HbA1c from baseline to the end of the study. Secondary efficacy endpoints included change in fasting plasma glucose, fasting plasma insulin and plasma lipids. At selected centres, oral glucose tolerance tests were performed and C-peptide and pro-insulin levels were measured. RESULTS: Mean HbA1c values decreased by the same amount in the two treatment groups from baseline to week 52 [pioglitazone: -1.4%; gliclazide: -1.4%; (90% CI: -0.18 to 0.02)]. A significantly greater mean reduction in fasting plasma glucose was observed in the pioglitazone group (2.4 mmol/l) than in the gliclazide group [2.0 mmol/l; treatment difference -0.4 mmol/l in favour of pioglitazone; P = 0.002; (95% CI: 0.7 to -0.1)]. Improvements in high-density lipoprotein cholesterol (HDL-C) and total cholesterol/HDL-C were greater with pioglitazone than with gliclazide (P < 0.001). The frequencies of adverse events were comparable between the two treatment groups, but more hypoglycaemic events were reported for gliclazide, whereas twice as many patients reported oedema with pioglitazone than with gliclazide. CONCLUSIONS: Pioglitazone monotherapy was equivalent to gliclazide in reducing HbA1c, with specific differences between treatments in terms of mechanism of action, plasma lipids and adverse events. PMID- 15787664 TI - Preserved insulin response to tolbutamide in hepatocyte nuclear factor-1alpha mutation carriers. AB - AIMS: Diabetic subjects with mutations in the gene encoding hepatocyte nuclear factor (HNF)-1alpha (MODY3) are prone to develop hypoglycaemia at low doses of glibenclamide, interpreted as sulphonylurea hypersensitivity. The present study was undertaken to compare the plasma insulin responses to glucose and tolbutamide in HNF-1alpha mutation carriers with those of healthy control subjects. METHODS: Seven mutation carriers; three normoglycaemic, two with impaired glucose tolerance, and two with newly detected diabetes, underwent an oral glucose tolerance test and a tolbutamide-modified intravenous glucose tolerance test with measurements of plasma insulin. Twenty-two healthy subjects served as controls. RESULTS: The plasma insulin response to intravenous glucose was reduced in the HNF-1alpha mutation carriers compared to the control subjects, with an area under the curve (median (interquartile range)) of 812 min pmol/l (421, 1647) and 1933 min pmol/l (1521, 2908), respectively (P = 0.03). In striking contrast, the plasma insulin response to tolbutamide was preserved, with an area under the curve of 2109 min pmol/l (1126, 3172) and 2250 min pmol/l (1614, 3276) in the mutation carriers and control subjects, respectively. CONCLUSIONS: HNF-1alpha mutation carriers are characterized by preserved tolbutamide-induced insulin secretion. Compared to healthy subjects, our MODY3 individuals did not show any increased serum insulin response to tolbutamide, suggesting that HNF-1alpha mutation carriers are not characterized by sulphonylurea hypersensitivity. PMID- 15787665 TI - Teaching and motivating patients to control their risk factors retards progression of cardiovascular as well as microvascular sequelae of Type 2 diabetes mellitus- a randomized prospective 8 years follow-up study. AB - AIMS: To examine whether motivating patients to gain expertise and closely follow their risk parameters will attenuate the course of microvascular and cardiovascular sequelae of diabetes. METHODS: A randomized prospective study on 165 patients with diabetes mellitus Type 2, hypertension (> 140/90 mmHg) and hyperlipidaemia (LDL-C > 3 mmol/l), referred for consultation to a diabetes clinic in an academic hospital. Patients were randomly allocated to standard consultation (SC) or to a patient participation (PP) and teaching programme. Follow-up continued by primary care physicians. RESULTS: The mean follow-up was 7.7 years. SC group patients each attended eight annual consultations. The PP patients initiated on average 1.2 +/- 0.8 additional consultations per annum. The relative risk (RR) over 8 years, for the combined cardiovascular event index in the intervention (PP) vs. the control (SC) group was 0.65 (95% CI 0.41-0.89, P = 0.001). Nephropathy developed in 14 vs. 7 patients in the SC and PP groups, respectively, RR 0.50 (95% CI 0.28-0.85, P = 0.02), retinopathy developed in 35 vs. 21 patients, RR 0.60 (95% CI 0.21-0.82, P = 0.03). Throughout the study, period blood pressure, LDL-C and HbA1c were significantly lower in the PP than in the SC patients. CONCLUSION: Well-informed and motivated patients, were more successful in maintaining good control of their risk factors, resulting in reduced cardiovascular risk and slower progression of microvascular disease. PMID- 15787666 TI - Role of habit in adherence to medical treatment. AB - This paper presents an analysis, from a philosophical point of view, of three psychological models describing how one adopts a health behaviour. More specifically, this analysis makes it possible to explain the role of habit in therapeutic adherence. By clarifying the notions of intention and decision, this novel analysis may be useful for understanding the dynamics of diabetes care. PMID- 15787667 TI - School attendance in children with Type 1 diabetes. AB - AIMS: To determine whether children with Type 1 diabetes mellitus (DM) miss more school than their non-DM siblings and peers and to identify factors associated with school absenteeism in children with DM. METHODS: School absenteeism data for the 2000-01 school year were obtained for 78 children with DM, 38 non-DM siblings and 118,269 age-matched peers in Toronto, Ontario. Questionnaires and hospital records were utilized to evaluate child-, family- and diabetes-related factors associated with school absenteeism in children with DM. RESULTS: Children with DM missed only slightly, albeit significantly more school than both their non-DM siblings (mean +/-sd: 10.9 +/- 8.9 vs. 8.1 +/- 8.1 days, P < 0.001) and peers (median: 8.8 vs. 5.5 days, P = 0.0005). A multiple regression analysis indicated that school absenteeism in children with DM was associated with their parents' attitudes towards school attendance (P = 0.002), poorer metabolic control (P = 0.006), shorter disease duration (P = 0.006) and a lack of aggressive behaviour (P = 0.02). CONCLUSIONS: With current management strategies, near normal school attendance is a reasonable goal for all children with DM and should be strongly encouraged by parents, educators and health care professionals. PMID- 15787668 TI - Is the relationship between adherence behaviours and glycaemic control bi directional at adolescence? A longitudinal cohort study. AB - AIM: To analyse the change in adherence to diabetes treatment and its association with metabolic control from childhood to adolescence. The Tanner pubertal staging model was selected as a marker of developmental maturity. METHODS: In a multicentre, longitudinal cohort study, 142 children with Type 1 diabetes completed a scale that assessed adherence to treatment and a test of diabetes knowledge at the beginning of the study (T0) and four years later (T4). HbA1c and clinical data were collected at T0 and at T4. RESULTS: From T0 to T4, the mean HbA1c increased from 8.2 +/- 1.6 to 9.1 +/- 1.4% (P < 0.001). Among patients at pubertal stages 1-4, adherence did not decline from T0 to T4, whereas the HbA1c level increased and a positive correlation between adherence and the knowledge score was noted (r = 0.32; P < 0.02). Among adolescents at pubertal stage 5, the level of adherence decreased (P < 0.01) from T0 to T4 and the HbA1c level increased despite an increase in the knowledge score (P < 0.001), a negative correlation between HbA1c and adherence was found (r = -0.37; P = 0.001) and adherence at T4 significantly added to the prediction of HbA1c at T4. CONCLUSIONS: In this longitudinal study, an initial worsening of glycaemic control as a result of puberty preceded worsening of adherence behaviours. Low levels of adherence become predictive of HbA1c degradation among pubertal stage 5 adolescents. These results suggest a potential bi-directional relationship between glycaemic control and adherence. PMID- 15787669 TI - Effect of maternal diabetes on phosphorylation of insulin-like growth factor binding protein-1 in cord serum. AB - AIMS: The insulin-like growth factor (IGF) system is considered important in the regulation of fetal growth. Binding of IGFs to specific binding proteins (IGFBPs) modifies their actions. In fetal blood, IGFBP-1 is the primary IGF binding protein whose phosphorylation generates proteins with different affinities for IGF-I. We studied cord serum IGFBP-1 phosphoisoform profiles in normal pregnancies and in diabetic pregnancies, which are frequently complicated by macrosomia. RESEARCH DESIGN AND METHODS: Cord serum IGFBP-1 phosphoisoform concentrations were measured at birth by two immunoenzymometric assays in 67 pregnancies complicated by Type 1 diabetes, in 28 pregnancies complicated by insulin-treated gestational diabetes, and in 62 normal pregnancies. RESULTS: Cord serum highly phosphorylated IGFBP-1 (hpIGFBP-1) concentrations were lower in pregnancies complicated by Type 1 diabetes (204 +/- 36 microg/l, P = 0.032) and in pregnancies complicated by gestational diabetes (170 +/- 28 microg/l, P = 0.031) than in controls (316 +/- 34 microg/l). Cord serum lesser phosphorylated IGFBP-1 (lpIGFBP-1) concentrations were similar in diabetic and normal pregnancies (P = 0.692 between groups by analysis of variance). Relative birth weight correlated negatively with cord serum hpIGFBP-1 and lpIGFBP-1 in diabetic pregnancies, and with cord serum lpIGFBP-1 in normal pregnancies. CONCLUSIONS: Maternal diabetes is associated with suppressed hpIGFBP-1 but unaltered lpIGFBP-1 concentrations in cord serum, suggesting that IGFBP-1 phosphoisoforms are differentially regulated in the fetus. Because hpIGFBP-1 has a higher affinity for IGF-I than does lpIGFBP-1, diabetes-related changes in fetal IGFBP-1 phosphorylation may increase IGF-I bioavailability and, consequently, stimulate fetal growth. This may partly explain the increased occurrence of macrosomia in diabetic pregnancies. PMID- 15787670 TI - Birthweight and other pregnancy outcomes in a cohort of women with pre gestational insulin-treated diabetes mellitus, Scotland, 1979-95. AB - AIMS: To assess pregnancy outcomes, in particular birthweight, in a large population-based cohort of women in Scotland with pre-gestational insulin-treated diabetes mellitus. METHODS: Data about diabetes from the Diabetes UK cohort were linked to data on births from the Scottish Hospital In-Patient Record System. This identified 1112 eligible singleton deliveries during 1979-95 to 706 insulin treated women. RESULTS: One thousand and eighty-four (97.5%) deliveries resulted in a live-born infant and 28 (2.5%) in a stillbirth. There were 13 (1.2%) neonatal deaths. The mean birthweight of the live-born infants was 3421 g, 1.06 standard deviations greater than that of infants in the Scottish general population after correcting for sex and gestational age. Forty-three per cent of live-born babies in the study were large (> Scottish 90th percentile) and 4% small (< 10th percentile) for their sex and gestational age. Macrosomia, defined as birthweight > or = 4000 g, occurred in 23% live-born babies and its prevalence was significantly inversely related to duration of maternal diabetes. However, the mean birthweight of infants born to mothers with diabetes for 20 or more years was still 0.90 standard deviations greater than in the general population. Prevalence of macrosomia increased with increasing number of previous pregnancies, but was not associated with maternal height or smoking habits. Stillbirth and neonatal death rates were, respectively, 4.7 (95% confidence interval = 3.3, 6.8) and 2.4 (1.4, 4.1), times higher than those in the general population. CONCLUSIONS: The frequency of adverse pregnancy outcomes in women with pre-existing insulin-treated diabetes was much higher than in the Scottish general population, and changed little during the study period. A detailed quantification of the independent effect of duration of mother's diabetes on birthweight revealed a continuous inverse correlation between these two variables. PMID- 15787671 TI - Autoantibodies to GAD and IA-2 in Saudi Arabian diabetic patients. AB - AIMS: To determine the prevalence of autoantibodies in sera of Saudi diabetic patients including Type 1 and Type 2 diabetes mellitus (DM) and gestational diabetes mellitus (GDM) living in Jeddah, Saudi Arabia. Apart from data on the prevalence of islet-cell antibodies in patients in Ryhadh (Al-Attas et al. Frequency of islet cell antibodies in adult newly diagnosed diabetic patients. Ann Saudi Med 1990; 10: 369-373) immunological markers of autoimmune diabetes have not been explored in Saudi Arabians. METHODS: Autoantibodies to GAD65 (GADA) and IA-2 (IA-2A) were determined using radio-immunoprecipitation assays. RESULTS: In Type 1 DM patients, 54% were GADA+ and 27% were IA-2A+. A greater negative effect of disease duration was noted for IA-2A than for GADA positivity. Autoantibodies were more prevalent with younger age of onset. GADA were slightly more common in female Type 1 DM patients. In Type 2 DM, 8/99 patients were GADA+, and three of these patients with shorter disease duration were also IA-2A+. GADA, and particularly IA-2A, were associated with a younger age of onset of Type 2 DM and all the autoantibody-positive Type 2 DM patients were insulin-treated. GADA were detected in 2.2% of GDM patients, but none of these patients possessed IA 2A. CONCLUSIONS: The prevalence and associations of autoantibodies in Saudi diabetic patients are very similar to those reported for diabetic patients in other ethnic groups. PMID- 15787672 TI - Increased renal glucose metabolism in Type 1 diabetes mellitus. AB - AIMS: In poorly controlled diabetes, increased renal glucose uptake has been implicated in the pathogenesis of diabetic nephropathy by promoting nonenzymatic glycosylation of proteins, activation of protein kinase C, and increased polyol pathway flux. However, whether glucose uptake by the diabetic kidney is actually increased, especially in patients with Type 1 diabetes, is unclear. METHODS: To examine this question, we used a combination of net balance and isotopic techniques to compare renal glucose uptake in 12 subjects with Type 1 diabetes before and after restoration of near normoglycaemia by infusion of insulin with that in 15 postabsorptive nondiabetic volunteers. RESULTS: Prior to insulin infusion, the diabetic subjects were markedly hyperglycaemic (arterial glucose 15.8 +/- 0.9 vs. 4.4 +/- 0.1 mm) and their renal tissue glucose uptake (i.e. total glucose disappearance across the kidney minus glycosuria) was increased more than 2 1/2-fold (388 +/- 43 vs. 148 +/- 12 micromol/min, P < 0.001). This was wholly explained by the mass action effects of hyperglycaemia since the diabetic subjects had normal renal blood flow (1575 +/- 82 vs. 1492 +/- 68 mL/min, P = 0.46) and reduced renal tissue glucose fractional extraction (1.7 +/- 0.2 vs. 2.3 +/- 0.1%, P = 0.027). Insulin infusion for three hours, which restored near normoglycaemia (arterial glucose 7.6 +/- 0.7 mm), reduced renal tissue glucose uptake toward normal (258 +/- 41 micromol/min, P = 0.006) without altering renal blood flow (1557 +/- 110, P = 0.63) or renal tissue glucose fractional extraction (2.1 +/- 0.3%, P = 0.35). Renal and hepatic glucose release, which had been increased (419 +/- 49 and 960 +/- 54 vs. 204 +/- 9 and 734 +/- 32 micromol/min, both P < 0.001), were suppressed by insulin to 138 +/- 22 and 520 +/- 53 micromol/min, respectively (both P < 0.001). CONCLUSIONS: In poorly controlled Type 1 diabetes, renal glucose uptake is markedly increased, which provides a link between hyperglycaemia and biochemical processes implicated in the pathogenesis of diabetic nephropathy. Its reversal by restoration of near normoglycaemia with insulin may explain the benefit of intensive insulin therapy in preventing diabetic nephropathy. PMID- 15787673 TI - Transcutaneous oxygen tension monitoring after successful revascularization in diabetic patients with ischaemic foot ulcers. AB - OBJECTIVE: To monitor transcutaneous oxygen tension (TcPO2) after percutaneous transluminal angioplasty (PTA) in diabetic patients with ischaemic foot ulcers. RESEARCH DESIGN AND METHODS: Twenty-three diabetic patients with ischaemic foot ulcers who underwent successful revascularization by PTA (SR group) were retrospectively selected. Twenty diabetic patients who underwent unsuccessful revascularization (UR group) were also included. Transcutaneous oxygen tension was measured at the dorsum of the foot before and 1 (+/- 1), 7 (+/- 1), 14 (+/- 1), 21 (+/- 1) and 28 (+/- 1) days after the surgical procedure. RESULTS: After PTA, TcPO2 progressively improved in the SR group, reaching its peak 4 weeks after angioplasty. A concomitant decrease of cutaneous carbon dioxide tension (TcPCO2) was also observed immediately after PTA which reached the lowest levels 3 weeks later. In the UR group, TcPO2 showed a slight improvement immediately after PTA but remained stable throughout the observation, while TcPCO2 levels did not change. Finally, the percentage of SR patients with a TcPO2 > or = 30 mmHg was 38.5% 1 week after PTA, while it increased to 75% 3 weeks later. CONCLUSION: Transcutaneous oxygen tension monitoring showed that after successful revascularization it takes 3-4 weeks for cutaneous oxygenation to improve and reach the optimal levels for wound healing. Transcutaneous carbon dioxide tension monitoring may be more useful to identify the negative outcome of a revascularization procedure. Our findings suggest that, when the surgical approach can be delayed, the best timing to perform a more aggressive debridement or minor amputations is 3-4 weeks after successful revascularization. PMID- 15787674 TI - Clinical characteristics of diabetic ketoacidosis in Korea over the past two decades. AB - AIMS: The aim of this study was to investigate changes in the clinical characteristics of diabetic ketoacidosis (DKA) in Korea over the last two decades. METHODS: A retrospective medical record review of all episodes of DKA from 1982 to 2002 in four University-affiliated urban hospitals in Korea was performed. A total of 255 episodes of DKA (217 patients) were identified and divided into three consecutive 7-year periods to compare trends over time. Clinical characteristics including precipitating factors and hospital mortality were analyzed. RESULTS: A dramatic increase in DKA admissions has occurred over the last two decades, accompanied by a marked increase in admissions of diabetic patients. The clinical characteristics of DKA remained constant over the observation period. Non-compliance to treatment was the most common precipitating factor of DKA. A total of 30 patients died in hospital (11.8% of all episodes). Older age and infection appeared to influence mortality. CONCLUSIONS: Our results suggest that rapidly increasing episodes of DKA in Korea, in parallel with increases in the numbers of diabetic patients, continue to be associated with significant mortality. PMID- 15787675 TI - alpha-Glucosidase inhibition (acarbose) fails to enhance secretion of glucagon like peptide 1 (7-36 amide) and to delay gastric emptying in Type 2 diabetic patients. AB - AIM: Acarbose is able to enhance GLP-1 release and delay gastric emptying in normal subjects. The effect of alpha-glucosidase inhibition on GLP-1 has been less evident in Type 2 diabetic patients. The aim of this study was to investigate the possible influence of acarbose on GLP-1 release and gastric emptying in Type 2 diabetic patients after a mixed test meal. PATIENTS AND METHODS: Ten Type 2 diabetic patients were tested with 100 mg acarbose or placebo served with a mixed meal that was labelled with 100 mg 13C-octanoic acid. Plasma concentrations of glucose, insulin, C-peptide, glucagon, GLP-1 and GIP were determined over 6 h. Gastric emptying was measured by determining breath 13CO2 using infrared absorptiometry. Statistics repeated-measures anova. RESULTS: Gastric emptying rates (t1/2: 162 +/- 45 vs. 163 +/- 62 min, P = 0.65) and plasma concentrations (increasing from approximately 12 to approximately 25 pmol/l, P = 0.37) and integrated responses of GLP-1 (P = 0.37) were not changed significantly by acarbose treatment. Postprandial plasma glucose concentrations (P < 0.0001) and their integrated responses were lowered by acarbose (by 64%; P = 0.016). The plasma concentrations of insulin and C-peptide were reduced (P = 0.007 and 0.057, respectively) by acarbose, while glucagon was not changed (P = 0.96). GIP plasma concentrations (increasing with placebo from approximately 10 to approximately 85 pmol/l and with acarbose to approximately 55 pmol/l (P < 0.0001) and their integrated responses were significantly lowered (by 43%) by acarbose (P = 0.021). After 2 weeks of acarbose treatment (50 mg t.i.d. for the first and 100 mg t.i.d. for the second week, n = 6), similar results were found. CONCLUSIONS: In hyperglycaemic Type 2 diabetic patients, ingestion of acarbose with a mixed test meal failed to enhance GLP-1 release and did not influence gastric emptying. PMID- 15787676 TI - Prevalence and risk factors for stroke in Type 2 diabetic patients in Taiwan: a cross-sectional survey of a national sample by telephone interview. AB - AIMS: To determine the prevalence and risk factors for stroke in patients with Type 2 diabetes mellitus (T2DM) and the age-specific prevalence odds ratios (POR) in comparison with the general population in Taiwan. METHODS: A total of 16 994 T2DM patients were randomly selected for telephone interview from a group covered by the National Health Insurance programme. Lifetime prevalence of stroke was calculated and various risk factors were analysed. Age-specific POR was calculated using previously reported prevalence of stroke in the general population from a nationwide survey across Taiwan. Standardized prevalence and POR were also calculated using the 2000-2025 population of the World Health Organization. RESULTS: A total of 12 531 cases (73.7%) were successfully interviewed. Stroke prevalence was 7.5%. In multivariate logistic regression, independent predictors were: increasing age, male gender, lower body mass index, ex-smokers, hyperlipidaemia, systolic pressure (or diastolic pressure when systolic pressure was not adjusted), education level below high school, and living in eastern or southern Taiwan. When compared with the general population, POR for stroke in the age groups < 45, 45-54, 55-64 and > or = 65 years were 82.29 (9.60, 705.57), 5.43 (2.33, 12.68), 3.73 (2.20, 6.33) and 2.14 (1.59, 2.89), respectively. The age-standardized prevalence of stroke was 2.3% in the diabetic patients and 0.6% in the general population. CONCLUSIONS: Stroke prevalence in Taiwanese T2DM is 7.5%. Diabetic patients have a higher risk of stroke than the general population, but the relative risk attenuates with age. Besides conventional atherosclerotic risk factors, stroke patients in Taiwan are characterized by lower body mass index, lower education level and residence in southern or eastern Taiwan. The negative association between body mass index and stroke in Taiwanese T2DM is in contrast to the generally accepted concept that obesity is a major risk factor as seen in most western countries. PMID- 15787677 TI - Does aerobic fitness influence microvascular function in healthy adults at risk of developing Type 2 diabetes? AB - AIM: To investigate whether aerobic fitness is associated with skin microvascular function in healthy adults with an increased risk of developing Type 2 diabetes. METHODS: Twenty-seven healthy normal glucose-tolerant humans with either a previous diagnosis of gestational diabetes or having two parents with Type 2 diabetes and 27 healthy adults who had no history of diabetes were recruited. Maximal oxygen uptake was assessed using an incremental exercise test to exhaustion. Skin microvascular function was assessed using laser Doppler techniques as the maximum skin hyperaemic response to a thermal stimulus (maximum hyperaemia) and the forearm skin blood flow response to the iontophoretic application of acetylcholine (ACh) and sodium nitroprusside. RESULTS: Maximal oxygen uptake was not significantly different in the 'at-risk' group compared with healthy controls. Maximum hyperaemia was reduced in those 'at risk' (1.29 +/ 0.30 vs. 1.46 +/- 0.33 V, P = 0.047); however, the peak response to acetylcholine or sodium nitroprusside did not differ in the two groups. A significant positive correlation was demonstrated between maximal oxygen uptake and maximum hyperaemia (r = 0.52, P = 0.006 l/min and r = 0.60, P = 0.001 ml/kg/min) and peak ACh response (r = 0.40, P = 0.04 l/min and r = 0.47, P = 0.013 ml/kg/min) in the 'at-risk' group when expressed in absolute (l/min) or body mass-related (ml/kg/min) terms. No significant correlations were found in the control group. CONCLUSIONS: In this 'at-risk' group with skin microvascular dysfunction maximal oxygen uptake was not reduced compared with healthy controls. However, in the 'at-risk' group alone, individuals with higher levels of aerobic fitness also had better microvascular and endothelial responsiveness. PMID- 15787678 TI - Criteria for previously undiagnosed diabetes and risk of mortality: 15-year follow-up of the Edinburgh Artery Study cohort. AB - AIMS: To compare risk of all-cause and cardiovascular mortality associated with different criteria for undiagnosed diabetes and glucose tolerance. METHODS: A population-based cohort of 758 men and 738 women of 55-74 years of age who had an oral glucose tolerance test or known diabetes at baseline were followed up until death or for 15 years. Mortality outcomes were compared by baseline diabetes status using people with normal glucose tolerance (i.e. those without diabetes, impaired fasting glucose or impaired glucose tolerance) as the reference group. RESULTS: Prevalence of undiagnosed diabetes using World Health Organization (WHO) criteria (fasting glucose of > or = 7.0 mmol/l and/or a 2-h post-challenge glucose of > or = 11.1 mmol/l) was 6.6%, of which 81% was associated with fasting glucose > or = 7.0 mmol/l and 19% was associated with isolated post-challenge hyperglycaemia. Hazard ratios (95% CI) for all-cause mortality adjusted for age and sex were 1.51 (1.09-2.08) for new diabetes by the American Diabetes Association (ADA) criterion (fasting glucose of > or = 7.0 mmol/l regardless of post-challenge glucose), 1.60 (1.20-2.13) for new diabetes by WHO criteria and 1.98 (1.14-3.44) for isolated post-challenge hyperglycaemia. Hazard ratios (95% CI) for cardiovascular mortality adjusted for age and sex were 1.89 (1.17-3.00), 1.73 (1.12-2.66) and 1.08 (0.34-3.40) for new diabetes by ADA and WHO criteria and for isolated post-challenge hyperglycaemia, respectively. CONCLUSIONS: Undiagnosed diabetes was associated with increased risk of all-cause mortality by any criteria but significantly increased cardiovascular disease mortality was only associated with diabetes diagnosed using the fasting glucose criterion. Mortality risks were similar in this population using either ADA or WHO criteria for diagnosis of diabetes. PMID- 15787679 TI - Reduced cardiovascular morbidity and mortality associated with metformin use in subjects with Type 2 diabetes. AB - AIM: Metformin therapy reduces microvascular complications in Type 2 diabetes; questions remain, however, regarding its impact on macrovascular events. This study examined metformin use in relation to risk of cardiovascular-related hospitalization and mortality. METHODS: We conducted a retrospective cohort analysis, using Saskatchewan Health administrative databases to identify new users of oral antidiabetic drugs. Subject groups were defined by medication use during 1991-1999: sulphonylurea monotherapy, metformin monotherapy, or combination therapy. Deaths and non-fatal hospitalizations recorded during the study period were identified as cardiovascular-related from ICD-9 codes. The main outcome was a composite of first non-fatal hospitalization or death. Standard multivariate techniques, including propensity scores, were used to adjust for potential confounding. Multivariate Cox proportional hazard models were used to examine the relationship between metformin use and the composite endpoint. RESULTS: Metformin monotherapy was associated with a lower risk of the composite endpoint (adjusted hazard ratio 0.81; 95% confidence interval 0.68, 0.97) compared with sulphonylurea monotherapy. Combination therapy with meformin and a sulphonylurea was associated with lower mortality, but had similar hospitalization rates, to sulphonylurea monotherapy. CONCLUSIONS: Metformin monotherapy was associated with a lower risk of cardiovascular-related morbidity and mortality, and combination metformin and sulphonylurea therapy was associated with a reduced risk of fatal cardiovascular events, when compared with sulphonylurea monotherapy. PMID- 15787680 TI - Dysfunction of the cyclo-oxygenase pathway in the foetoplacental circulation in Type 1 diabetes mellitus. AB - AIM: In diabetes, perinatal morbidity is significantly increased. This may partly be related to functional changes in the foetoplacental vascular bed. In diabetes models, a defect in the cyclo-oxygenase pathway is a common observation. Therefore, we hypothesized that the human foetoplacental circulation of diabetic patients is characterized by dysfunction of the cyclo-oxygenase pathway, as well. METHODS: We performed ex-vivo perfusions of isolated placental cotyledons from healthy women (n = 14) and from patients with Type 1 diabetes (n = 9). The contribution of cyclo-oxygenase products to foetoplacental vascular tone was quantified by measuring the response to the cyclo-oxygenase inhibitor indomethacin. RESULTS: Baseline foetoplacental arterial pressure was comparable between controls and diabetic women (mean +/- sem, 21.7 +/- 1.2 vs. 24.4 +/- 2.0 mmHg). Maximum foetoplacental arterial pressure at highest dose of indomethacin was 32.8 +/- 3.0 mmHg in controls vs. 27.3 +/- 2.3 mmHg in diabetic women. The indomethacin-induced increase in pressure was reduced in diabetes (2.9 +/- 0.7 vs. 11.2 +/- 2.4 mmHg in controls, P = 0.01). CONCLUSIONS: Under baseline conditions, the net effect of all cyclo-oxygenase products in the foetoplacental vascular bed is vasodilation. In diabetes, this vasodilator effect seems significantly impaired. PMID- 15787681 TI - The introduction of a specific request form for the diagnosis of gestational diabetes (GDM) improves understanding of GDM amongst clinicians but does not increase its detection. PMID- 15787682 TI - Association between fasting glucose and C-reactive protein in middle-aged subjects. PMID- 15787683 TI - Prognostic value of admission plasma glucose and HbA in acute myocardial infarction. PMID- 15787685 TI - A new cause of neuroglycopenia: 'missing the point'-impaired visual function during acute hypoglycaemia. PMID- 15787687 TI - In response to 'variations in glucose self-monitoring during oral hypoglycaemic therapy in primary care'. PMID- 15787690 TI - Brief exposure to NMDA produces long-term protection of cerebellar granule cells from apoptosis. AB - Cerebellar granule cells (CGCs) require excitatory inputs to survive during their postnatal migration from the external to the internal granule cell layers. The lack of innervation of mossy fibres induces CGC death by apoptosis. In vitro, CGCs die by apoptosis in the presence of physiological concentrations of KCl (5 mm or K5) but they survive in the presence of depolarizing concentrations of KCl (25 mm or K25) or N-methyl-d-aspartate (NMDA) by a mechanism dependent on calcium influx. The addition of NMDA or K25, for only 24 h, to immature CGCs cultures [2 days in vitro (DIV)] was able to produce a remarkable and long-term protection until 8 DIV. Moreover, our data show that NMDA and K25-mediated long-lasting protection was related to an inhibition of caspase-3 activity. By using different protein kinase inhibitors, we have shown that the inhibition of caspase-3 activation by NMDA was dependent on the activation of tyrosine kinases and phosphatidylinositol 3-kinase (PI3-kinase). Moreover, an impairment in NMDA mediated neuroprotection and caspase-3 inhibition was observed when the action of brain-derived neurotrophic factor (BDNF) was blocked. By contrast, K25-mediated neuroprotection was BDNF-independent and was mediated by a mitogen-activated protein kinase- and PI3-kinase-dependent inhibition of caspase-3. PMID- 15787691 TI - A modified MPTP treatment regime produces reproducible partial nigrostriatal lesions in common marmosets. AB - Standard MPTP treatment regimens in primates result in > 85% destruction of nigral dopaminergic neurons and the onset of marked motor deficits that respond to known symptomatic treatments for Parkinson's disease (PD). The extent of nigral degeneration reflects the late stages of PD rather than events occurring at its onset. We report on a modified MPTP treatment regimen that causes nigral dopaminergic degeneration in common marmosets equivalent to that occurring at the time of initiation of motor symptoms in man. Subcutaneous administration of MPTP 1 mg/kg for 3 consecutive days caused a reproducible 60% loss of nigral tyrosine hydroxylase (TH)-positive cells, which occurred mainly in the calbindin-D(28k) poor nigrosomes with a similar loss of TH-immunoreactivity (TH-ir) in the caudate nucleus and the putamen. The animals showed obvious motor abnormalities with reduced bursts of activity and the onset of motor disability. However, the loss of striatal terminals did not reflect early PD because a greater loss of TH-ir occurred in the caudate nucleus than in the putamen and a marked reduction in TH ir occurred in striatal patches compared to the matrix. Examination of striatal fibres following a partial MPTP lesion showed a conspicuous increase in the number and the diameter of large branching fibres in the putaminal and to some extent caudatal matrix, pointing to a possible compensatory sprouting of dopaminergic terminals. In addition, these partially lesioned animals did not respond to acute treatment with L-DOPA. This primate partial lesions model may be useful for examining potential neuroprotective or neurorestorative agents for PD. PMID- 15787692 TI - A combination drug therapy improves cognition and reverses gene expression changes in a mouse model of Huntington's disease. AB - Huntington's disease is a genetic disease caused by a single mutation. It is characterized by progressive movement, emotional and cognitive deficits. R6/2 mice transgenic for exon 1 of the HD gene with 150+ CAG repeats have a progressive neurological phenotype, including deterioration in cognitive function. The mechanism underlying the cognitive deficits in R6/2 mice is unknown, but dysregulated gene expression, reduced neurotransmitter levels and abnormal synaptic function are present before the cognitive decline becomes pronounced. Our goal here was to ameliorate the cognitive phenotype in R6/2 mice using a combination drug therapy (tacrine, moclobemide and creatine) aimed at boosting neurotransmitter levels in the brain. Treatment from 5 weeks of age prevented deterioration in two different cognitive tasks until at least 12 weeks. However, motor deterioration continued unabated. Microarray analysis of global gene expression revealed that many genes significantly up- or down-regulated in untreated R6/2 mice had returned towards normal levels after treatment, though a minority were further dysregulated. Thus dysregulated gene expression was reversed by the combination treatment in the R6/2 mice and probably underlies the observed improvements in cognitive function. Our study shows that cognitive decline caused by a genetic mutation can be slowed by a combination drug treatment, and gives hope that cognitive symptoms in HD can be treated. PMID- 15787693 TI - 19-Nortestosterone influences neural stem cell proliferation and neurogenesis in the rat brain. AB - Abuse of androgenic anabolic steroids can affect brain function leading to behavioural changes. In this study, the effects of the testosterone analogue, 19 nortestosterone, on rat neural stem cells was examined. The androgen receptor is expressed by cultured embryonic and adult neural stem cells, and is also present in the ventricular epithelium during development and in the adult brain in, among others, dentate gyrus. In neural stem cells stimulated with epidermal growth factor, nandrolone reduced cell proliferation, especially in adult ones. The decrease was abolished by flutamide, a receptor antagonist. Nandrolone also decreased the BrdU labelling of neural stem cells in the dentate gyrus, demonstrating an effect of the hormone on cell proliferation in vivo. The effect of nandrolone was observed with both female and male rats but it was more pronounced in pregnant rats, indicating an involvement of oestrogen in nandrolone action. Nandrolone also decreased the number of newly born neuronal cells in the dentate gyrus of male rats. The results show that nandrolone has important effects on the proliferation and differentiation of neural stem cells expressing the cognate androgen receptor. The data show that the use of nandrolone may severely affect the formation of neural stem cells and could therefore have long term negative consequences in the brain. PMID- 15787694 TI - Effects of Abeta1-42 fibrils and of the tetrapeptide Pr-IIGL on the phosphorylation state of the tau-protein and on the alpha7 nicotinic acetylcholine receptor in vitro. AB - In order to investigate the possible links connecting beta-amyloid (Abeta) accumulation, tau-hyperphosphorylation and nicotinic receptor expression, rat embryonic primary hippocampal cultures were incubated with amyloidogenic peptides. Exposure to 0.5 microm fibrillar Abeta(1-42) for 3 days caused retraction of dendrites, shrinkage of cell bodies and a decrease in the expression of microtubule-associated proteins 2b (MAP2b), without affecting the total number of neurons and their viability. No impact on the tau-phosphorylation sites Ser-202, Thr231/Ser235, Ser262 and Ser396/Ser404 was found. The total number of homomeric alpha7-nicotinic receptors (alpha7-nAChRs) and their affinity for [(125)I]alpha-bungarotoxin remained unaltered. Upon incubation with the putatively protective tetrapeptide propionyl-isoleucine-isoleucine-glycine leucine (Pr-IIGL), an analogue of the region [31-34] of Abeta, cell bodies were swollen in the region of the apical dendrite. These morphological alterations, different from those elicited by Abeta(1-42), did not involve MAP2 expression changes. In contrast to Abeta(1-42), Pr-IIGL caused a massive hyperphosphorylation of the tau-protein at Ser-202 and at Ser396/Ser404. The total number of homomeric alpha7-nAChRs and their affinity for [(125)I]alpha bungarotoxin were unaffected. In conclusion, the present results show a toxic effect of Abeta(1-42) on the cytoskeletal structure at concentrations normally present in the brains of Alzheimer's disease patients, but raise some doubts about the role of Abeta(1-42) fibrils as a direct trigger of tau hyperphosphorylation. The tetrapeptide Pr-IIGL cannot be considered protective with regard to cell morphology. Although it prevents the Abeta(1-42)-induced retraction of dendrites, it exhibits other toxic properties. The homomeric alpha7 nAChRs were not affected either by Abeta(1-42) incubation or by Pr-IIGL-induced tau-hyperphosphorylation. PMID- 15787695 TI - Differential effects of forskolin on tyrosine hydroxylase gene transcription in identified brainstem catecholaminergic neuronal subtypes in organotypic culture. AB - The regulation of gene expression of tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis, was studied in brainstem noradrenergic nuclei, locus coeruleus (LC), A2 and A1, in vitro. Several novel experimental approaches employed in this study included: (i) the development of a slice-explant model in which these brainstem nuclei maintained a high survival of the noradrenergic neurons, an organotypic topology and the coexpression of two identifying markers in addition to TH, i.e. norepinephrine transporter (NET) and vesicular monoamine transporter 2 (VMAT2); (ii) quantitative analysis of TH transcription in these nuclei was made using a labelled intronic probe to measure TH heteronuclear RNA (hnRNA) and (iii) the use of tetrodotoxin in the media to eliminate spontaneous neural activity in these nuclei, thereby providing a basal state as the starting point for the study of TH transcription under various pharmacological perturbations. In the presence of TTX, the adenylcyclase stimulator, forskolin, produced a 155% increase in LC, a 130% increase in A1, and a 220% increase in A2 in TH hnRNA as compared to control nuclei. This effect of forskolin was abolished in the LC and A1 by the PKA inhibitor, H89 (5 microm), but not by the MAP kinase pathway (MEK) inhibitor, PD98059 (75 microm). In contrast, the robust increase in TH transcription produced by forskolin in A2 neurons, was completely inhibited by PD98059, and only partially inhibited by H89, showing that induced TH transcription is mediated by different kinase pathways in specific central noradrenergic neuronal subtypes. PMID- 15787696 TI - Distinct properties of functional KCC2 expression in immature mouse hippocampal neurons in culture and in acute slices. AB - A hallmark in the development of GABAergic neurotransmission is the switch in GABA(A)-mediated responses from depolarizing to hyperpolarizing. This occurs due to a gradual decrease in the intracellular concentration of chloride caused by the functional expression of the neuron-specific K-Cl cotransporter KCC2. Whether a mere increase in the amount of KCC2 protein is the rate-limiting step in vivo, or a further activation of the otherwise nonfunctional cotransporter is required, is not clear. Imposing a fixed Cl(-) load via patch pipette we measured the resultant somato-dendritic gradients in reversal potential of GABAergic currents to determine the time course of functional maturation of KCC2-mediated Cl(-) extrusion in two preparations: cultured mouse hippocampal neurons plated at embryonic day 17 and CA1 pyramidal cells in acute slices. We found that in immature neurons in both preparations the gradient is initially small or not detectable. It undergoes an abrupt increase at around days 13-14 in culture, while a more gradual increase occurs between postnatal days 5-14 in slices. Consistent with the presence of a nonfunctional form of KCC2 in immature hippocampal neurons grown in culture, application of the broad-spectrum kinase inhibitor staurosporine produces a rapid and potent up-regulation of KCC2 function in these cultured neurons, but not in neonatal slices. Taken together with our previously published data, these results indicate that the functional activity of KCC2 in vivo parallels the developmental expression of the protein, whereas cultured neurons require an additional activation step (mimicked by staurosporine) for KCC2 to become functional. PMID- 15787697 TI - Investigation of PKC isoform-specific translocation and targeting of the current of the late afterhyperpolarizing potential of myenteric AH neurons. AB - AH neurons in the enteric nervous system play an essential role in initiating intestinal reflexes and factors that control AH neuron excitability therefore influence the state of the digestive system. Prominent afterhyperpolarizations that follow action potentials in these neurons strongly affect their excitability. In the present work, we have investigated the regulation of the afterhyperpolarizing current (I(AHP)) by protein kinase C (PKC). Electrophysiological responses and protein translocation were investigated in AH neurons of freshly dissected preparations of myenteric ganglia from the guinea pig ileum. The activator of conventional and novel PKCs, phorbol dibutyrate, but not the activator of novel PKCs, ingenol, blocked the I(AHP). Phorbol dibutyrate had no effect on the hyperpolarization-activated current (I(h)) or on the A current (I(A)). Stimulation of synaptic inputs to the neurons also reduced the I(AHP), and had no effect on I(h) or I(A). Phorbol dibutyrate also reduced a background outward current that was present after the I(AHP) current had been blocked by clotrimazole. Both phorbol dibutyrate and ingenol caused translocation of the novel PKC, PKCepsilon, in these neurons. Only phorbol dibutyrate caused translocation of PKCgamma, a conventional PKC. The studies thus indicate that the activation of PKC by phorbol esters and by nerve stimulation affects AH neurons in a similar way, and that PKC activation targets both the I(AHP) and another background K(+) current. The I(AHP) is targeted by a conventional PKC, suggested to be PKCgamma, as this is the only conventional PKC that is prominent in AH neurons. PMID- 15787698 TI - Homogeneous distribution of large-conductance calcium-dependent potassium channels on soma and apical dendrite of rat neocortical layer 5 pyramidal neurons. AB - Voltage-gated conductances on dendrites of layer 5 pyramidal neurons participate in synaptic integration and output generation. We investigated the properties and the distribution of large-conductance calcium-activated potassium channels (BK channels) in this cell type using excised patches in acute slice preparations of rat somatosensory cortex. BK channels were characterized by their large conductance and sensitivity to the specific blockers paxilline and iberiotoxin. BK channels showed a pronounced calcium-dependence with a maximal opening probability of 0.69 at 10 microm and 0.42 at 3 microm free calcium. Their opening probability and transition time constants between open and closed states are voltage-dependent. At depolarized potentials, BK channel gating is described by two open and one closed states. Depolarization increases the opening probability due to a prolongation of the open time constant and a shortening of the closed time constant. Calcium-dependence and biophysical properties of somatic and dendritic BK channels were identical. The presence of BK channels on the apical dendrite of layer 5 pyramidal neurons was shown by immunofluorescence. Patch clamp recordings revealed a homogeneous density of BK channels on the soma and along the apical dendrite up to 850 microm with a mean density of 1.9 channels per microm(2). BK channels are expressed either isolated or in clusters containing up to four channels. This study shows the presence of BK channels on dendrites. Their activation might modulate the shape of sodium and calcium action potentials, their propagation along the dendrite, and thereby the electrotonic distance between the somatic and dendritic action potential initiation zones. PMID- 15787699 TI - Dynamics of learning-induced spine redistribution along dendrites of pyramidal neurons in rats. AB - We have previously shown that olfactory-discrimination (OD) learning is accompanied by enhanced spine density along proximal apical dendrites of layer II pyramidal neurons in the piriform (olfactory) cortex. Here we studied the temporal dynamics of learning-induced modifications in dendritic spine density throughout the dendritic trees of these neurons. We observed a transient increase in proximal apical spine density after OD learning, suggesting a strengthening of intrinsic excitatory inputs interconnecting neurons within the olfactory cortex. By contrast, the afferent pathway receiving direct input from the olfactory bulb shows spine pruning, suggesting that the connectivity is weakened. The changes in spine density can be attributed to a net change in number of spines, as the morphometric parameters of the dendrites are unaffected by learning. We suggest that spine density changes may represent a mechanism of selective synaptic reorganization required for olfactory learning consolidation. PMID- 15787700 TI - Stabilizing effects of extracellular ATP on synaptic efficacy and plasticity in hippocampal pyramidal neurons. AB - The role of adenosine triphosphate (ATP) as a neurotransmitter and extracellular diffusible messenger has recently received considerable attention because of its possible participation in the regulation of synaptic plasticity. However, the possible contribution of extracellular ATP in maintaining and regulating synaptic efficacy during intracellular ATP depletion is understudied. We tested the effects of extracellular ATP on excitatory postsynaptic currents (EPSCs) evoked in CA1 pyramidal neurons by Schaffer collateral stimulation. In the absence of intracellular ATP, EPSC rundown was neutralized when a low concentration of ATP (1 microm) was added to the extracellular solution. Adenosine and ATP analogues did not prevent the EPSC rundown. The P(2) antagonists piridoxal-5'-phosphate azophenyl 2',4'-disulphonate (PPADS) and reactive blue-2, and the P(1) adenosine receptor antagonist 8-cyclopentyltheophylline (CPT) had no detectable effects in cells depleted of ATP. However, the protective action of extracellular ATP on synaptic efficacy was blocked by extracellular application of the protein kinase inhibitors K252b and staurosporine. In contrast, K252b and staurosporine per se did not interfere with synaptic transmission in ATP loaded cells. Without intracellular ATP, bath-applied caffeine induced a transient (< 35 min) EPSC potentiation that was transformed into a persistent long-term potentiation (> 80 min) when 1 microm ATP was added extracellularly. An increased probability of transmitter release paralleled the long-term potentiation induced by caffeine, suggesting that it originated presynaptically. Therefore, we conclude that extracellular ATP may operate to maintain and regulate synaptic efficacy and plasticity in conditions of abnormal intracellular ATP depletion by phosphorylation of a surface protein substrate via activation of ecto-protein kinases. PMID- 15787701 TI - Somatodendritic autoreceptor regulation of serotonergic neurons: dependence on L tryptophan and tryptophan hydroxylase-activating kinases. AB - The somatodendritic 5-HT(1A) autoreceptor has been considered a major determinant of the output of the serotonin (5-HT) neuronal system. However, recent studies in brain slices from the dorsal raphe nucleus have questioned the relevance of 5-HT autoinhibition under physiological conditions. In the present study, we found that the difficulty in demonstrating 5-HT tonic autoinhibition in slice results from in vitro conditions that are unfavorable for sustaining 5-HT synthesis. Robust, tonic 5-HT(1A) autoinhibition can be restored by reinstating in vivo 5-HT synthesizing conditions with the initial 5-HT precursor l-tryptophan and the tryptophan hydroxylase co-factor tetrahydrobiopterin (BH(4)). The presence of tonic autoinhibition under these conditions was revealed by the disinhibitory effect of a low concentration of the 5-HT(1A) antagonist WAY 100635. Neurons showing an autoinhibitory response to L-tryptophan were confirmed immunohistochemically to be serotonergic. Once conditions for tonic autoinhibition had been established in raphe slice, we were able to show that 5 HT autoinhibition is critically regulated by the tryptophan hydroxylase activating kinases calcium/calmodulin protein kinase II (CaMKII) and protein kinase A (PKA). In addition, at physiological concentrations of L-tryptophan, there was an augmentation of 5-HT(1A) receptor-mediated autoinhibition when the firing of 5-HT cells activated with increasing concentrations of the alpha(1) adrenoceptor agonist phenylephrine. Increased calcium influx at higher firing rates, by activating tryptophan hydroxylase via CaMKII and PKA, can work together with tryptophan to enhance negative feedback control of the output of the serotonergic system. PMID- 15787702 TI - The relationship between V6 and PO in macaque extrastriate cortex. AB - The cerebral cortex of three macaque monkeys, electrophysiologically studied in chronic preparations in order to recognize functionally the medial parieto occipital area V6, was reconstructed using the software CARET. Locations of cells recorded from area V6 (n = 553) and from neighbouring cortical areas V2/V3 and V6A (n = 1341) were displayed on surface-based reconstructions of individual brains, and on a surface-based atlas of the macaque cerebral cortex. Results show that area V6 occupies the ventral part and fundus of the parieto-occipital sulcus, as well as the ventral part of the precuneate cortex. V6 borders areas V2/V3 posteriorly and laterally, and area V6A anteriorly. The visualization of individual cases on a common template (atlas), and the use of atlas datasets, allowed us to compare data coming from different individuals and different laboratories. In particular, a comparison of the location and extent of the medial parieto-occipital areas V6 and PO indicates that area PO occupies different locations according to different authors but in general includes parts of both areas V6 and V6A. We therefore suggest that the term V6 is a more appropriate designation of the visuotopically-organized area located on the anterior wall of the parieto-occipital sulcus. PMID- 15787703 TI - Correlation between multiple climbing fibre regression and parallel fibre response development in the postnatal mouse cerebellum. AB - At the neuromuscular junction elimination of supernumerary synaptic connections during development is owing to a competitive process between single neuronal populations, whereas in the central nervous system the interaction of different types of input could affect the process. In the cerebellum, the regression from multiple- to mono-innervation of the Purkinje cells by climbing fibres is virtually completed during the first two weeks of postnatal development. While it is clear that parallel fibres are important in the control of the regression, there are conflicting results in relation to whether an early phase of regression is independent of parallel fibre effects. We studied the precise timing of climbing fibre synapse development and decline and the relationship with the functional maturation of parallel fibres. Until postnatal day (P) 6 or 7, the synaptic currents generated by different climbing fibres become progressively more uniform in amplitude. However, between P7 and 14, the amplitudes of the currents increasingly diverge until only one fibre remains connected. These data are taken as evidence that, in multiply innervated Purkinje cells, competition between different climbing fibres appears at P7 and continues during the second postnatal week. Morphological and electrophysiological data demonstrate that parallel fibres synapses appear at P7 and their development is significantly correlated with the time course of the climbing fibre regression. These results provide no evidence for climbing fibre regression independent of parallel fibres before P7 and also suggest a dominant role of the parallel fibre input in the later phase. PMID- 15787704 TI - Purkinje cell loss affects differentially the execution, acquisition and prepulse inhibition of skeletal and facial motor responses in Lurcher mice. AB - Adult heterozygous Lurcher mice show a degeneration of almost all Purkinje cells and 90% of the granular cells of the cerebellum, resulting in ataxia or general deficits in motor coordination. These mice are therefore an excellent model for studying the role of the cerebellar cortex in motor performance, including the acquisition of new motor abilities. The performance of 3-month-old Lurcher mice was studied in various behavioural (fall, horizontal bar, rotating cylinder, and ladder), spatial orientation (water maze) and associative learning (eyelid classical conditioning) tasks and compared with that of wild-type mice. Behavioural tasks indicated a deficit for motor abilities in Lurcher mice but with some adaptation to the tests and improvement in performance. Wild-type and Lurcher mice performed swimming equally, but the latter learned the task significantly more slowly than the former. The late component of reflex blinks was smaller in amplitude and had a longer latency in Lurcher mice than in controls. Learning curves for Lurcher mice during classical conditioning of eyelid responses were similar to controls, but the amplitude of the learned response in Lurcher mice was significantly lower. The startle response to a severe tone was similar in both control and Lurcher mice but the latter were unable to produce prepulse inhibition. These results suggest that the cerebellar cortex is not indispensable for the performance of this complete set of skeletal and facial tasks, or for the acquisition of new motor abilities, but it is for the appropriate execution and adjustment of any of these motor activities. PMID- 15787705 TI - Bi-hemispheric contribution to functional motor recovery of the affected forelimb following focal ischemic brain injury in rats. AB - In many recovering hemiparetic stroke patients, movement of the affected limb elicits ipsilateral activation of sensorimotor areas within the undamaged hemisphere, which is not observed in control subjects. Following middle cerebral artery occlusion, rats received intensive enriched-rehabilitation (ER) of the impaired forelimb for 4 weeks. Weekly assessments on a skilled reaching test demonstrated significant improvement in ischemic animals over 4 weeks of ER (P < 0.05). We hypothesized that if the undamaged forelimb motor cortex contributed to improved forelimb function, then inhibition of neural activity within this region should reinstate (at least some of) the initial motor impairment. After 3 and 4 weeks of ER, animals received a microinjection of lidocaine hydrochloride into the undamaged motor cortex and were re-assessed on reaching ability. The behavioral effect of lidocaine challenge was dependent on the size of the infarct: animals with large infarcts were rendered unable to retrieve any food pellets and had great difficulty even contacting a pellet with the affected forepaw. Small-infarct animals were only moderately affected (25% reduction in success) by lidocaine, an effect similar to that observed in control animals. Qualitative assessments of recovered reaching after 4 weeks of rehabilitation revealed that impairments in forelimb lift, advance and aim were exacerbated (P < 0.05) following lidocaine-inactivation of the undamaged motor cortex of animals with large ischemic infarcts. In animals with small infarcts, lidocaine challenge only impaired limb advance. Thus, recruitment of the undamaged hemisphere may depend on the functional integrity of the remaining sensorimotor system. These data suggest that, in the rat, the undamaged (ipsilateral) motor system may contribute to compensatory recovery of the affected forelimb. PMID- 15787706 TI - Exposure to pulse-modulated radio frequency electromagnetic fields affects regional cerebral blood flow. AB - We investigated the effects of radio frequency electromagnetic fields (RF EMF) similar to those emitted by mobile phones on waking regional cerebral blood flow (rCBF) in 12 healthy young men. Two types of RF EMF exposure were applied: a 'base-station-like' and a 'handset-like' signal. Positron emission tomography scans were taken after 30 min unilateral head exposure to pulse-modulated 900 MHz RF EMF (10 g tissue-averaged spatial peak-specific absorption rate of 1 W/kg for both conditions) and sham control. We observed an increase in relative rCBF in the dorsolateral prefrontal cortex on the side of exposure. The effect depended on the spectral power in the amplitude modulation of the RF carrier such that only 'handset-like' RF EMF exposure with its stronger low-frequency components but not the 'base-station-like' RF EMF exposure affected rCBF. This finding supports our previous observation that pulse modulation of RF EMF is necessary to induce changes in the waking and sleep EEG, and substantiates the notion that pulse modulation is crucial for RF EMF-induced alterations in brain physiology. PMID- 15787707 TI - Divergence and convergence of thalamocortical projections to premotor and supplementary motor cortex: a multiple tracing study in the macaque monkey. AB - The premotor cortex of macaque monkeys is currently subdivided into at least six different subareas on the basis of structural, hodological and physiological criteria. To determine the degree of divergence/convergence of thalamocortical projections to mesial [supplementary motor area (SMA)-proper and pre-SMA] and lateral (PMd-c, PMd-r, PMv-c and PMv-r) premotor (PM) subareas, quantitative analyses were performed on the distribution of retrograde labelling after multiple tracer injections in the same animal. The results demonstrate that all PM and SMA subareas receive common inputs from several thalamic nuclei, but the relative contribution of these nuclei to thalamocortical projections differs. The largest difference occurs between subareas of SMA, with much greater contribution from the mediodorsal (MD) and area X, and a smaller contribution from the ventral lateral anterior (VLa) and ventral part of the ventral lateral posterior (VLpv) to pre-SMA than to SMA-proper. In PM, differences between subareas are less pronounced; in particular, all receive a significant contribution from MD, the ventral anterior (VApc) and area X. However, there are clear gradients, such as increasing projections from MD to rostral, from VLa and VLpv to caudal, and from dorsal VLp (VLpd) to dorsal premotor subareas. Intralaminar nuclei provide widespread projections to all premotor subareas. The degree of overlap between thalamocortical projections varies among different PM and SMA subareas and different sectors of the thalamus. These variations, which correspond to different origin and topography of thalamocortical projections, are discussed in relation to functional organizations at thalamic and cortical levels. PMID- 15787708 TI - Modulation of synaptic transmission in neocortex by network activities. AB - Neocortical neurons integrate inputs from thousands of presynaptic neurons that fire in vivo with frequencies that can reach 20 Hz. An important issue in understanding cortical integration is to determine the actual impact of presynaptic firing on postsynaptic neuron in the context of an active network. We used dual intracellular recordings from synaptically connected neurons or microstimulation to study the properties of spontaneous and evoked single-axon excitatory postsynaptic potentials (EPSPs) in vivo, in barbiturate or ketamine xylazine anaesthetized cats. We found that active states of the cortical network were associated with higher variability and decrease in amplitude and duration of the EPSPs owing to a shunting effect. Moreover, the number of apparent failures markedly increased during active states as compared with silent states. Single axon EPSPs in vivo showed mainly paired-pulse facilitation, and the paired-pulse ratio increased during active states as compare to silent states, suggesting a decrease in release probability during active states. Raising extracellular Ca(2+) concentration to 2.5-3.0 mm by reverse microdialysis reduced the number of apparent failures and significantly increased the mean amplitude of individual synaptic potentials. Quantitative analysis of spontaneous synaptic activity suggested that the proportion of presynaptic activity that impact at the soma of a cortical neuron in vivo was low because of a high failure rate, a shunting effect and probably dendritic filtering. We conclude that during active states of cortical network, the efficacy of synaptic transmission in individual synapses is low, thus safe transmission of information requires synchronized activity of a large population of presynaptic neurons. PMID- 15787709 TI - alpha-Amino-3-hydroxy-5-methylisoxazole-4-propionate receptor autoradiography in mouse brain after single and repeated withdrawal from diazepam. AB - Withdrawal from chronic treatment with benzodiazepines is associated with increased neuronal excitability leading to anxiety, aversive effects and increased seizure sensitivity. After repeated withdrawal experiences, seizure sensitivity increases while withdrawal-induced anxiety and aversion decrease. We used autoradiographical methods employing [(3)H]Ro48 8587, a selective ligand for glutamatergic alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors, to study withdrawal-induced changes in AMPA receptor binding in areas of the mouse brain postulated to be involved in these responses. Mice were given 21 days treatment with diazepam (15 mg/kg, s.c. in sesame oil) followed by withdrawal (single withdrawal) or three blocks of 7 days treatment interspersed with 3-day periods to allow washout of drug (repeated withdrawal). In keeping with heightened excitability in withdrawal from chronic diazepam treatment, the single withdrawal group showed, 72 h after their final dose of diazepam, increased [(3)H]Ro48 8587 binding in several brain areas associated with emotional responses or seizure activity, including hippocampal subfields, amygdalar and thalamic nuclei and motor cortex. In contrast, the repeated withdrawal group showed no changes in [(3)H]Ro48 8587 binding in any brain area studied. These observations are consistent with up-regulation of AMPA receptor mediated transmission being important in withdrawal-induced anxiety and aversion but not in increased seizure sensitivity associated with repeated withdrawal. As changes in AMPA receptor subunit expression alter the functionality of the receptor, future studies will address this possibility. PMID- 15787711 TI - PDE10A inhibition reverses subchronic PCP-induced deficits in attentional set shifting in rats. AB - Persistent suppression of N-methyl-d-aspartate (NMDA) receptor function produces enduring structural changes in neocortical and limbic regions in a pattern similar to changes reported in schizophrenia. This similarity suggests that chronic NMDA receptor antagonism in animals may represent a useful model of neurobiological and related cognitive deficits in schizophrenia. Schizophrenia is associated with impairments in frontal lobe-dependent cognitive functions, including working memory and attentional shifting. Deficits in attention and executive function have not been well characterized in animal models of schizophrenia using chronic NMDA receptor antagonist administration. We investigated whether subchronic systemic administration of the NMDA receptor antagonist phencyclidine (PCP) to rats followed by a drug washout period would produce enduring cognitive deficits on an attentional set-shifting task. The task is functionally analogous to a sensitive test of frontal function in humans and non-human primates. Subchronic PCP administration selectively impaired extradimensional shift learning without affecting other discrimination or reversal tasks. Moreover, acute treatment with the PDE10A inhibitor papaverine immediately prior to testing attenuated the PCP-induced deficits in extradimensional shift learning across a range of doses. These data suggest that subchronic PCP administration may model effectively some of the cognitive deficits that are observed in schizophrenia, and that PDE10A inhibition may be an effective therapeutic route to improve executive function deficits associated with schizophrenia. PMID- 15787710 TI - Inhibition of restraint stress-induced neural and behavioural activation by endogenous cannabinoid signalling. AB - The role of endocannabinoid (eCB) signalling in restraint stress-induced neuronal activation was studied. Male mice exposed to 30 min of restraint exhibit increased Fos protein within prefrontal cortex (PFC), lateral septum (LS), nucleus accumbens (Acb) and medial amygdala. SR141716 (2 mg/kg) itself had no effect on Fos but pretreatment with SR141716 significantly potentiated restraint induced Fos expression in cingulate, LS and Acb. SR141716 also significantly increased the time spent in active escape behaviours during the restraint. In restraint-habituated mice (mice exposed to four previous restraint episodes), the fifth restraint exposure resulted in decreased expression of active escape behaviours compared to the first exposure and only induced Fos protein in the central and medial amygdala. Administration of SR141716 prior to the fifth restraint episode resulted in greater potentiation of restraint-induced Fos induction than the first; significant increases occurred within all regions of PFC examined, LS and Acb. Brain regional eCB content was measured immediately after restraint. N-arachidonylethanolamine content within the amygdala was significantly decreased after both restraint episodes. 2-Arachidonylglycerol content was significantly increased in both the limbic forebrain and amygdala after the fifth restraint but not the first. Restraint had no effect on cerebellar eCB content. These data suggest that eCB activation of CB(1) receptors opposes the behavioural and neuronal responses to aversive stimuli. Because repeated homotypic stress increased both limbic 2-AG and resulted in a greater effect of SR141716 on limbic Fos expression, we hypothesize that increased CB(1) receptor activity contributes to the expression of habituation to homotypic stress. PMID- 15787712 TI - REM sleep: a sensitive index of fear conditioning in rats. AB - To examine the influence of conditioned fear stimuli on sleep-wake states, we recorded sleep in Sprague-Dawley rats after exposure to tones previously paired with footshock. After habituation to a recording chamber and the recording procedure, a baseline sleep recording was obtained the next day. One day later, experimental animals were exposed to shock training designed to induce conditioned fear (FC), consisting of five tone-footshock pairings. The 5-s tones (conditioned stimuli; CS) co-terminated with 1-s footshocks (unconditioned stimuli; US). The next day sleep was recorded for 4 h in the recording chamber after presentation of five CSs alone. Sleep efficiency (total sleep time/recording period) and REM sleep (REM) and non-REM (NREM) measures were determined. While sleep efficiency was not significantly changed after CS presentation, the percentage of total sleep time spent in REM (REM percentage) was reduced in the FC animals. The reduction in REM percentage in the FC animals was due to a decrease in the number of REM bouts. In a separate experiment, we repeated the procedures, except the tones and shocks were presented in an explicitly unpaired (UP) fashion. The next day, presentation of the tones increased REM percentage in the UP group. Results are discussed in terms of the decreases in REM as a response to conditioned fear, and the relevance of these findings to the sleep changes seen in post-traumatic stress disorder (PTSD). PMID- 15787713 TI - Anterior perirhinal cortex kindling produces long-lasting effects on anxiety and object recognition memory. AB - Temporal lobe epilepsy (TLE) is frequently accompanied by memory impairments and, although their bases are unknown, most research has focused on the hippocampus. The present study investigated the importance of another medial temporal lobe structure, the perirhinal cortex (Prh), in changes in memory in TLE using kindling as a model. Rats were kindled twice daily with anterior Prh stimulation until three fully generalized seizures were evoked. Beginning 7 days later and on successive days, rats were tested in an elevated plus maze, a large circular open field, an open field object exploration task and a delayed-match-to-place task in a water maze in order to assess anxiety-related and exploratory behaviour, object recognition memory and spatial cognition. Kindling increased anxiety-related behaviour in both the elevated plus and open field mazes and disrupted spontaneous object recognition but spared all other behaviours tested. These results are consistent with other findings indicating a greater role for the Prh in object memory and emotional behaviour than in spatial memory and contrast with the selective disruption of spatial memory produced by dorsal hippocampal kindling. The site-selectivity of the behavioural disruptions produced by kindling indicates that such effects are probably mediated by changes particular to the site of seizure initiation rather than to changes in the characteristic circuitry activated by limbic seizure generalization. Further investigation of the behavioural effects of Prh kindling may be useful for studying the mechanisms of mnemonic and affective dysfunction associated with TLE and offer insights into bases for variability in such dysfunction across patients. PMID- 15787714 TI - Neurons in monkey visual cortex detect lines defined by coherent motion of dots. AB - Form perception from coherent motion is an important aspect of vision. Representations of one-, two- and three-dimensional forms have been found at various stages of cortical processing using random-dot stimuli, whereas representations of biological objects like a walking human being concentrate at higher stages of processing. The perception of biological objects can be induced by sparse dot stimuli that consist of a few dots that mark the joints of the human body [G. Johansson (1973) Percept. Psychophys., 14, 201-211]. In the present study, we aimed to investigate whether neurons in early visual areas that respond to bars and edges defined by luminance contrast also signal bar-like objects from sparse dot stimuli. We studied single neurons with rows of 3-24 dots that were either collinear or scattered within a rectangular form. These dots were moved coherently on a uniform or dotted background, and human observers perceived them as rigid rods or other bar-like objects. We found neurons in the visual cortex of the awake, behaving monkey that responded to these stimuli and were sensitive to the orientation of these objects as for conventional bars or edges. Stimulus conditions that failed to induce these percepts in human observers also evoked weaker responses or none in these neurons. We found these neurons with increasing frequency in areas V1, V2 and V3/V3A. The results suggest that the visual cortex not only detects biological objects, but also lines and other bar-like objects from sparse dot stimuli, and that this function evolves at an early stage of processing. PMID- 15787715 TI - The neural correlate of very-long-term picture priming. AB - Repetition priming denotes a behavioural change caused by prior exposure to a stimulus. The effect is known to last for weeks. This study addresses the underlying neural mechanisms for very-long-term picture priming by using event related functional magnetic resonance imaging complemented by a behavioural paradigm. Previous functional imaging studies with shorter retention intervals have shown that priming is associated with changes in the activity of both the occipital and posterior temporal cortex. In this study we compared retention intervals of 1 day and 6 weeks after initial exposure to a picture stimulus. Priming-related decreases in cortical activity in posterior extrastriate and dorsal left inferior frontal areas were found only for the shorter retention interval. In contrast, fMRI activation in the inferior posterior temporal and anterior left inferior frontal cortex was reduced following priming for both retention intervals. In the behavioural paradigm, the priming effect was stable over time. We conclude that the left inferior frontal and inferior posterior temporal cortex play a key role in the very-long-term priming effect. PMID- 15787716 TI - Role of primary visual cortex in the binocular integration of plaid motion perception. AB - This study assessed the early mechanisms underlying perception of plaid motion. Thus, two superimposed gratings drifting in a rightward direction composed plaid stimuli whose global motion direction was perceived as the vector sum of the two components. The first experiment was aimed at comparing the perception of plaid motion when both components were presented to both eyes (dioptic) or separately to each eye (dichoptic). When components of the patterns had identical spatial frequencies, coherent motion was correctly perceived under dioptic and dichoptic viewing condition. However, the perceived direction deviated from the predicted direction when spatial frequency differences were introduced between components in both conditions. The results suggest that motion integration follows similar rules for dioptic and dichoptic plaids even though performance under dichoptic viewing did not reach dioptic levels. In the second experiment, the role of early cortical areas in the processing of both plaids was examined. As convergence of monocular inputs is needed for dichoptic perception, we tested the hypothesis that primary visual cortex (V1) is required for dichoptic plaid processing by delivering repetitive transcranial magnetic stimulation to this area. Ten minutes of magnetic stimulation disrupted subsequent dichoptic perception for approximately 15 min, whereas no significant changes were observed for dioptic plaid perception. Taken together, these findings suggest that V1 is not crucial for the processing of dioptic plaids but it is necessary for the binocular integration underlying dichoptic plaid motion perception. PMID- 15787717 TI - Attending to visual or auditory motion affects perception within and across modalities: an event-related potential study. AB - The present event-related potential (ERP) study examined the role of dynamic features in multisensory binding. It was tested whether endogenous attention to the direction of motion affects processing of visual and auditory stimuli within and across modalities. Human participants perceived horizontally moving dot patterns and sounds that were presented either continuously (standards) or briefly interrupted (infrequent deviants). Their task was to detect deviants moving in a particular direction within a primary modality, but to detect all deviants irrespective of their motion direction within the secondary modality. Attending to the direction of visual motion resulted in a broad selection negativity (SN) starting at about 200 ms post-stimulus onset, and attending to the direction of auditory motion resulted in a positive difference wave at 150 ms that was followed by a broad negativity starting at about 200 ms (unimodal effects). Moreover, dot patterns moving in a direction that was attended within audition were detected faster and more accurately than oppositely moving stimuli and elicited a cross-modal SN wave. Corresponding cross-modal behavioural and ERP results were obtained for sounds moving in a direction that was attended within vision. Unimodal and cross-modal ERP attention effects partially differed in their scalp topography. The present study shows that dynamic features (direction of motion) may be used to link input across modalities and demonstrates for the first time that these multisensory interactions take place as early as about 200 ms after stimulus onset. PMID- 15787718 TI - Increases in extracellular levels of 5-HT and dopamine in the basolateral, but not in the central, nucleus of amygdala induced by aversive stimulation of the inferior colliculus. AB - Consistent evidence has shown that dopamine release in the prefrontal cortex is increased by electrical stimulation of the inferior colliculus (IC) as unconditioned stimulus. Recent reports have also demonstrated that inactivation of the basolateral nucleus of the amygdala (BLA) with muscimol enhances the behavioural consequences of the aversive stimulation of the IC and reduces the dopamine release in the prefrontal cortex. Moreover, neurotoxic lesions of the BLA enhance whereas those of the central nucleus of the amygdala (CeA) reduce the aversiveness of the electrical stimulation of the IC. Based on these findings the present study examined the effects of the electrical stimulation of the IC on the extracellular levels of serotonin and dopamine in the BLA and CeA. To this end, rats implanted with a stimulation electrode in the IC also bore a microdialysis probe in the BLA or CeA for determination of the release of dopamine and serotonin. IC electrical stimulation at the freezing and escape thresholds increased the levels of serotonin ( approximately 70%) and dopamine ( approximately 60%) in the BLA related to the basal values. Similarly, the metabolites DOPAC and 5-HIAA increased in a parallel fashion in BLA. No significant changes could be detected in these biogenic amines and metabolites in CeA following IC aversive stimulation. These findings point to a differential role of serotonergic and dopaminergic mechanisms of the BLA and CeA in the setting up of adaptive responses to fear states generated at the inferior colliculus level. PMID- 15787719 TI - Semantic memory retrieval: cortical couplings in object recognition in the N400 window. AB - To characterize the regional changes in neuronal couplings and information transfer related to semantic aspects of object recognition in humans we used partial-directed EEG-coherence analysis (PDC). We examined the differences of processing recognizable and unrecognizable pictures as reflected by changes in cortical networks within the time-window of a determined event-related potential (ERP) component, namely the N400. Fourteen participants performed an image recognition task, while sequentially confronted with pictures of recognizable and unrecognizable objects. The time-window of N400 as indicative of object semantics was defined from the ERP. Differences of PDC in the beta-band between these tasks were represented topographically as patterns of electrical couplings, possibly indicating changing degrees of functional cooperation between brain areas. Successful memory retrieval of picture meaning appears to be supported by networks comprising left temporal and parietal regions and bilateral frontal brain areas. PMID- 15787720 TI - Nomenclature for factors of the HLA system, 2004. PMID- 15787721 TI - Anti-HLA class II antibodies in kidney retransplant patients. AB - The relevance of anti-HLA class II antibodies for kidney graft survival is still controversial. In part, this can be attributed to difficulties to detect and differentiate anti-HLA class II antibodies. Anti-HLA class II IgG antibody screening was performed by enzyme-linked immunosorbent assay. Subsequently, all anti-HLA class II-positive sera were subjected to the determination and specification using color-coded microspheres coated with purified HLA antigens. In a cohort of 934 patients awaiting kidney transplantation, 41 sera (4.4%) were positive for IgG anti-HLA class II antibodies. The presence was confirmed in 90.2% sera by retesting. Subsequently, all anti-HLA class II-positive patients (n = 27) who in the past had undergone a kidney transplantation with an HLA-DR and/or -DQ-mismatched graft were selected. In 25 of 27 sera (92.6%), the alloantibody specificities corresponded to the known previous transplant mismatches on a broad antigen level. In 20 of 27 sera (74.1%), anticlass I antibodies were detected as well. Anti-HLA-DP antibodies were seen in 24 of the 27 sera of this cohort. In the majority of the cases, the reactivities with different DPB1 alleles could be explained by involvement of a single, specific DPB1 epitope. Donor-specific anti-HLA-DR and -DQ antibodies were seen in the majority of cases with graft failure following HLA class II alloantigen exposure in prior kidney transplantations. In addition, HLA-DP may serve as a transplantation antigen in kidney transplantation, leading to a humoral response. PMID- 15787722 TI - Origin of Aymaras from Bolivia and their relationship with other Amerindians according to HLA genes. AB - Aymara Amerindians from the Titicaca Lake Andean highlands are studied for HLA-A, HLA-B, HLA-DRB1 and HLA-DQB1 gene frequencies. Genetic distances, neighbour joining and correspondence analyses are performed by using other Amerindian and worldwide populations (15384 chromosomes are studied). The HLA genetic profile of Aymaras is different from neighbouring and language-related Quechuas (Incas). Both Quechuas and Aymaras seem to present an HLA-DRB1*0901 high frequency, which is present in a very low frequency or absent in Mesoamericans (Mazatecans, Mayans) and most studied Amerindians. Moreover, it is observed a closer relatedness of Aymaras with Amerindians from the Amazon Basin and Chaco lowlands, compared to Quechuans. PMID- 15787723 TI - Identification and frequency of a new HLA-A allele, A*030104. AB - We here identified a novel HLA-A allele, A*030104, which was found in a Japanese family. The direct sequencing revealed that A*030104 was identical to A*030101 except for a nucleotide substitution of GAG to GAA at codon 63 without an amino acid replacement. The frequencies of A*030104 and A*0301 including A*030104 in Japanese population (n = 22,360) were approximately 0.013 and 0.40%, respectively. PMID- 15787724 TI - Identification of a novel HLA-DMB allele (DMB*0107) in the Korean population. AB - A novel allele for human leukocyte antigen-DMB was identified in the Korean population. DMB*0107 was identical to DMB*0101 at exon 2, apart from three mismatches at nucleotide positions 82 (A-->G), 146 (A-->T) and 212 (G-->A). These mutations resulted in codon changes at positions 10 (Thr-->Ala), 31 (Asp-->Val) and 53 (Ser-->Asn). PMID- 15787725 TI - Current methods of assessing platelet function: relevance to transfusion medicine. PMID- 15787726 TI - In vitro function of platelet concentrates prepared after filtration of whole blood or buffy coat pools. AB - BACKGROUND/METHOD: Data on the quality of platelet concentrates (PC) produced by the buffy coat method and stored beyond 5 days in plasma are limited. We therefore evaluated the quality of PCs prepared by leucocyte depletion of whole blood (Terumo WBSP, n = 10) or a buffy coat pool (Pall Autostop, n = 10), and stored for 7 days in plasma by assessing platelet parameters and markers of platelet activation. RESULTS: In both types of PC, levels of glucose decreased during storage but were not totally depleted (> 11 mM on day 7). In contrast, lactate levels increased on storage and was consistently < 20 mM throughout, with pH maintained at > 6.8 in all units. Hypotonic shock response scores were > 47% in all units at day 7. On day 1, markers of platelet activation were significantly higher in WBSP PC, but by day 7 were similar for percentage CD63+ and CD62P + (40%) with levels of platelet microparticles and annexin V binding two-fold higher in WBSP. The expression of CD61 did not alter during storage and the percentage of platelets expressing CD42b was > 88% in all units on day 7. RANTES (Regulated on activation, normal, T-cell expressed and secreted) and TGFbeta released from platelets by day 7 was < 800 ng/ml and 90 ng/ml, respectively. C3a(desarg) increased throughout storage in both types of PC, but without a commensurate increase in the terminal complex SC5b-9 or activation of factor XII. CONCLUSION: Our data indicates that the in vitro characteristics of PCs prepared using these methods is maintained over storage for 7 days in plasma and is not associated with significant deterioration of platelet function. ONE SENTENCE SUMMARY: In vitro function of platelet concentrates prepared by either filtration of whole blood, or pooled buffy coats. PMID- 15787727 TI - Occurrence of cold activation of transfusion plasma during storage at +4 degrees C. AB - BACKGROUND AND OBJECTIVES: The storage of transfusion plasma at +4 degrees C sometimes leads to the activation of several proteolytic systems. In this study the frequency of cold activation was investigated, as well as whether cold activation of plasma is an individually recurrent property of the donor. MATERIALS AND METHODS: Plasma units prepared from whole blood obtained from 100 male donors were stored at +2 degrees to +5 degrees C, in bags for 28 days and in cryotubes for up to 42 days. Samples from plasma units, collected by apheresis from 100 male donors, were stored in cryotubes for up to 42 days. Cold activation was measured weekly as kallikrein-like activity of plasma. Samples from repeat apheresis plasma units from 32 donors were measured 12-20 months later. The effects of storage on the contact, coagulation and fibrinolytic systems were determined. RESULTS: The cumulative frequency of cold-activated plasma units stored in bags was 5% on day 7 and 18% on day 28. After 42 days in cryotubes, 49% of the plasma units were cold activated. Large intraindividual differences in the onset-day of cold activation were observed in plasma samples of some donors. During cold activation, an increase in kallikrein-like activity was accompanied by a decrease in C1 esterase inhibitor activity and an increase in the concentrations of activated factor VII and fibrinopeptide A. The functional plasminogen level was unchanged, while a minor decrease in plasmin inhibitor activity was combined with a corresponding increase in the concentration of plasmin-plasmin inhibitor complex. CONCLUSIONS: The cumulative frequency of cold activated plasma units increased in a time-dependent manner during storage at +2 degrees to +5 degrees C for 42 days. The intraindividual onset-day of cold activation varied widely between plasma samples of some donors. Cold activation was associated with a high degree of activation of the contact and coagulation systems. The fibrinolytic system was scarcely affected. PMID- 15787728 TI - Diagnosis and procedure-specific survival among transfusion of recipients in 1993 and 2000, Orebro County, Sweden. AB - BACKGROUND AND OBJECTIVES: Survival rates in patients transfused in 1993 and 2000 were compared in relation to diagnoses and surgical interventions. MATERIALS AND METHODS: Blood centre and hospital records of all patients transfused from March to May in Orebro County in 1993 (n = 932) and 2000 (n = 990), were matched with the national register of deaths. RESULTS: Relative risk of death within 1 year, adjusted for diagnoses, operations and other confounders in patients transfused in 2000 compared to 1993 was 0.78 (CI 0.66-0.91). Among those transfused 1993, 39% were alive after 7 years. CONCLUSION: The improved survival among those transfused in 2000 could not be accounted for by differences in ages or case-mix. PMID- 15787729 TI - A prospective trial on the safety of long-term intensive plasmapheresis in donors. AB - BACKGROUND AND OBJECTIVES: There are still concerns about the safety of long-term intensive donor plasmapheresis, because the reasons that donors drop out of plasmapheresis programmes have not been determined in prospective studies. MATERIALS AND METHODS: Seventy-two donors were switched from a moderate plasmapheresis programme to an intensive plasmapheresis programme and observed over a 3-year period. In addition to measuring total serum protein (TSP), albumin, immunoglobulin G (IgG) and haemoglobin (Hb) levels, parameters of iron metabolism and blood coagulation, and biochemical cardiovascular risk markers, were determined at baseline and at every 15th donation. We also collected statements from donors who dropped out of the plasmapheresis programme about their reasons for withdrawal. RESULTS: Dropouts were predominantly related to socioeconomic (n = 34) or medical reasons not related to plasma donations (n = 8). Three donors had to drop out when their TSP levels fell below threshold for the third time within a 5-week period. At baseline, donors had significantly lower TSP, albumin, IgG, Hb and ferritin levels than gender-matched and age matched non-donor controls. However, subsequent intensive plasmapheresis over 3 years did not impair the individuals' ability to donate plasma. TSP, IgG, Hb, ferritin, transferrin, cardiovascular risk markers and parameters of blood coagulation did not change significantly during the observation period. CONCLUSIONS: The reasons why donors cease to participate in intensive plasmapheresis programmes are predominantly not directly related to the plasma donation itself. PMID- 15787730 TI - Identification of a novel A2 allele derived from the A transferase gene through a nucleotide substitution G539C. AB - BACKGROUND AND OBJECTIVES: The A2 is a very rare phenotype in the ABO blood group system in the Oriental population. It corresponds to a special ABO allele encoding a glycosyltransferase that is capable of synthesizing A2 antigens, which is weaker than the typical A antigen. In this study, we report a novel A2 allele in two unrelated Taiwanese individuals. MATERIALS AND METHODS: Two individuals were identified as the A2 phenotype based on the standard ABO serological test. For analysing the A2 allele, both direct sequencing and gene cloning of the ABO gene were performed. RESULTS: The ABO gene of the two A2 individuals was composed of O1 and A2 alleles, and the novel A2 allele has a 539G > C that results in the amino acid change Arg180Pro. The mutation was not detected in the general group A population. CONCLUSION: We report for the first time that a 539G > C mutation represents a new molecular basis for the A2 blood type. The amino acid substitution from arginine to proline may have effect on the expression of A antigen. PMID- 15787731 TI - Transfusion-transmitted malaria: current donor selection guidelines are not sufficient. AB - A case of transfusion-transmitted malaria was identified in a 50-year-old male patient with sickle cell disease. The donor was Ghanaian, but had migrated to the UK some years previously and had not left the UK for 8 years. The donor met all of the extant donor selection guidelines [1] and a donation was consequently collected. However, on subsequent investigation of the case, the donor was found to be parasitaemic and have high titre malarial antibodies. As a result of this case, changes to the United Kingdom donor selection guidelines have been proposed. These changes will prevent any such further transmissions. PMID- 15787732 TI - World Health Organization collaborative study to establish a replacement WHO international standard for hepatitis C virus RNA nucleic acid amplification technology assays. AB - BACKGROUND AND OBJECTIVES: A collaborative study was undertaken to establish a replacement for the current (1st) World Health Organization (WHO) hepatitis C virus (HCV) International Standard, 96/790. MATERIALS AND METHODS: Both the 1(st) International Standard and the replacement standard were prepared from the same starting material by diluting a high titre genotype 1a HCV isolate in pooled, human plasma. The only difference was that each standard was lyophilized in two, separate lyophilisation runs but under the same conditions. RESULTS: In the study to establish the 1st International Standard, no significant difference in potency was found between the material eventually designated as the 1st International Standard and that now selected as the 2nd International Standard. The present study also showed no significant differences between the materials stored at -20 degrees C and no evidence of degradation over 5 years. CONCLUSIONS: Material 96/798 was established as the 2nd HCV International Standard and assigned the same unitage as the 1st International Standard, i.e. 10(5) IU/ml (50,000 IU/vial). PMID- 15787733 TI - Hepatitis C test results do not justify claim of test equivalence nor of prolonged seroconversion. PMID- 15787734 TI - The rights of blood recipients should supersede any asserted rights of blood donors: the right to donate blood in South Africa. PMID- 15787735 TI - Transfusion of apheresis platelets and ABO groups. PMID- 15787738 TI - Meconium aspiration syndrome induces complement-associated systemic inflammatory response in newborn piglets. AB - The pathophysiology of meconium aspiration syndrome (MAS) is complex. We recently showed that meconium is a potent activator of complement. In the present study, we investigated whether the complement activation occurring in experimental MAS is associated with a systemic inflammatory response as judged by granulocyte activation and cytokine and chemokine release. MAS was induced by the instillation of meconium into the lungs of newborn piglets (n = 8). Control animals (n = 5) received saline under otherwise identical conditions. Haemodynamic and lung dynamic data were recorded. Complement activation, revealed by the terminal sC5b-9 complex (TCC), and cytokines [interleukin (IL)-6 and IL-8] were measured in plasma samples by enzyme immunoassays. The expression of CD18, CD11b and oxidative burst in granulocytes was measured in whole blood by flow cytometry. Plasma TCC increased rapidly in the MAS animals in contrast with controls (P < 0.0005). The TCC concentration correlated closely with oxygenation index (r = 0.48, P < 0.0005) and ventilation index (r = 0.57, P < 0.0005) and inversely with lung compliance (r = -0.63, P < 0.0005). IL-6 and IL-8 increased in MAS animals compared with the controls (P = 0.002 and P < 0.001, respectively). Granulocyte oxidative burst declined significantly in the MAS animals compared with the controls (P < 0.02). TCC correlated significantly with IL-6 (r = 0.64, P < 0.0005) and IL-8 (r = 0.32; P = 0.03) and inversely with oxidative burst (r = -0.37; P = 0.02). A systemic inflammatory response associated with complement activation is seen in experimental MAS. This reaction may contribute to the pathogenesis of MAS. PMID- 15787739 TI - Microparticles shed from different antigen-presenting cells display an individual pattern of surface molecules and a distinct potential of allogeneic T-cell activation. AB - Various cells such as platelets, lymphocytes, endothelial cells, red blood cells and monocytes do release surface-derived microparticles (mps). We analysed mp isolated from supernatants of cultured antigen-presenting human cells (APCs) and human cell lines. Particle sizing by dynamic light scattering revealed a characteristic size of the particles ranging from 80 nm to 300 nm in viable cells and from 400 nm to 1200 nm in irradiated cells. Employing flow-cytometry, we observed partly an altered surface protein composition of the mp compared to their cellular source. Mp originating from dendritic cells (DCs) differed in their surface composition from those released from monocytes and monocyte-derived macrophages. In functional assays, these mp stimulated alloreactive T-cells. The treatment of the cells with either UV-B or lipopolysaccharide strongly influenced the quantity, the immunostimulatory features and the surface composition of the mp. Mp from apoptotic macrophages were able to reduce the stimulatory capacity of vital macrophages but not of DC. Apoptotic mps from DC, on the other hand, were always stimulatory. This is the first report regarding the study of mp released from DC and compared with those released from other APC. PMID- 15787740 TI - Inhibition by Artocarpus tonkinensis of the development of collagen-induced arthritis in rats. AB - Extracts of the leaves and roots from the tree Artocarpus tonkinensis A Cheval (family Moraceae) are used in traditional Vietnamese medicine in order to treat backache as well as rheumatic joint diseases. We prepared an ethyl acetate (EtOAc) extract from this plant and tested its anti-inflammatory properties in an experimental arthritis model, collagen-induced arthritis (CIA). CIA was induced in Dark Agouti rats by means of immunization with collagen type II (CII) emulsified in incomplete Freund's adjuvant. Starting at the day of immunization, the rats were treated daily with intraperitoneal injections of Artocarpus extract. Arthritis progression was measured by means of clinical scoring of paws and anti-CII antibody titres were measured by means of ELISA. In vitro, lymph node (LN) cell cultures were treated with Artocarpus extract and the apoptosis inducing effect was determined with FACS staining by using annexin V and propidium iodide as well as the TUNEL method. Treatment of the rats with Artocarpus extract decreased arthritis incidence and severity and delayed disease onset. When treatment was started after the onset of arthritis, a tendency towards arthritis amelioration was observed. In vitro, Artocarpus extract acted as a T-cell modulator, inhibiting mitogen-induced T-cell proliferation and inducing apoptosis of activated LN-derived lymphocytes. Thus, we have demonstrated that an EtOAc extract of Artocarpus, a plant traditionally used in Vietnamese folk medicine for treating arthritic conditions, has beneficial effects in an experimental arthritis model. This effect is likely to be T cell dependent and mediated through apoptosis induction in activated cells. PMID- 15787741 TI - Mycobacterium avium ssp. paratuberculosis recombinant heat shock protein 70 interaction with different bovine antigen-presenting cells. AB - Abstract Heat shock proteins (Hsp) can deliver antigen into the major histocompatibility complex class I presentation pathway of antigen-presenting cells (APC), a process called cross priming, thus stimulating antigen-specific CD8+ T-cell reactions. Hsp were shown to elicit proinflammatory responses in APC. Both processes require interaction of Hsp with APC via specific receptors. This study describes the interaction of recombinant Hsp70 (rHsp70) of Mycobacterium avium subspecies paratuberculosis with bovine peripheral blood mononuclear cells that was restricted to CD14+ cells. Characterized monocyte-derived macrophages, monocyte-derived dendritic cells (DC) and BoMac, an immortalized bovine macrophage cell line, were used to investigate the interaction of rHsp70 with different bovine APC. Saturation of immature DC with high concentrations of rHsp70 is demonstrated, and it was found that interaction of rHsp70 with DC was related to the maturation stage of the DC. Involvement of CD91 as a cellular receptor for rHsp70 was demonstrated; however, competition studies with immature DC demonstrated that other receptors exist on bovine APC. These data suggest that rHsp70-based vaccines may be useful for the successful immunization of cattle. PMID- 15787742 TI - Influence of interleukin-4 on the phenotype and function of bone marrow-derived murine dendritic cells generated under serum-free conditions. AB - Murine bone marrow-derived dendritic cells (DC) can be generated by culture in the presence of granulocyte/macrophage colony-stimulating factor (GM-CSF) alone or GM-CSF in conjunction with interleukin-4 (IL-4). However, these two culture methods result in the production of heterogeneous DC populations with distinct phenotypic and stimulatory properties. In this study, we investigated the properties of DC generated under serum-free conditions in the presence or absence of IL-4 and compared their yield and phenotype to that of DC generated in the presence of fetal calf serum (FCS) (+/-IL-4). We did not observe a significant difference in the total cell yield between these four culture conditions, although the proportion of CD11c+ DC in cultures that received FCS was higher than that of their counterparts generated under serum-free conditions. Also, the four culture conditions generated CD11c+ DC with comparable levels of major histocompatibility complex (MHC) class II, CD40, CD80 and CD86 expression, with the exception of cells cultured under serum-free conditions in the absence of IL 4, which displayed suboptimal levels of these markers. Moreover, we compared the functional and stimulatory properties of DC generated under serum-free conditions in the presence or absence of IL-4. DC cultured in the presence of IL-4 were stronger stimulators of allogeneic splenocytes in a primary mixed lymphocyte reaction (MLR) and of naive antigen-specific OT-II transgenic T cells when pulsed with the class II ovalbumin (OVA)323-339 peptide or whole OVA protein than DC cultured in the absence of IL-4. However, both DC populations displayed a similar capacity to take up fluorescein isothiocyanate (FITC)-albumin by macropinocytosis and FITC-Dextran by the mannose receptor and to secrete IL-12 in response to stimulation with lipopolysaccharide (LPS) or an agonistic anti-CD40 monoclonal antibody. Therefore, we conclude that although both DC culture methods result in the production of DC with similar functional abilities, under serum-free conditions, DC cultured in GM-CSF and IL-4 show an increased stimulatory potential over DC cultured in GM-CSF alone. This is an important consideration in the design of experiments where DC are being exploited as immunotherapeutic vaccines. PMID- 15787743 TI - The inhibitory effect of altered collagen II peptide on HLA-DRB1-restricted T cell activation. AB - It has been known that rheumatoid arthritis (RA)-associated antigenic peptides CII263-272 are coupled with human leucocyte antigen (HLA)-DRB1 and recognized by T-cell receptor (TCR), which in turn induced T-cell proliferation and pathogenesis of RA. Non-T-cell-stimulating type II collagen (CII) peptides might be generated by removing the amino acids responsible for TCR contact and keeping the HLA-DR-binding residues intact. In this study, a panel of altered CII peptides (APs) with consecutive or single substitutions of the TCR-contacting residues were synthesized. Through peptide binding and T-cell activation assays, we demonstrated that altered CII263-272 peptides with substitution of the TCR contacting residues did not or barely induced T-cell activation; one of the best non-T-cell-stimulating peptide AP268-270 inhibited the binding of wild-type CII263-272 to HLA-DR1 and T-cell activation triggered by wild-type CII263-272 and HA306-318 in a dose-response manner. These data suggest that removal of the TCR contacting residues of CII263-272 leads to HLA-DRB1 binding and low T-cell stimulating peptides, which could potentially inhibit the T-cell response induced by HLA-DRB1-binding antigenic peptides. PMID- 15787744 TI - A combination of anergic cells' adoptive transfer and rapamycin therapy prolongs cardiac allograft survival in mice. AB - The in vivo immunoregulatory effect of anergic cells induced by blocking the costimulatory pathway was investigated in this study. Anergic cells were generated in vitro by mixed culture of murine splenic cells from BALB/c and C3H/HeJ under the blockade of anti-CD154 and anti-CD80 monoclonal antibodies, and the in vitro activity of anergic cells were observed. The 3.0 Gy gamma-irradiated BALB/c mice received cardic allografts from C3H/HeJ, and anergic cells were intravenously injected immediately after transplantation. Recipient mice injected with anergic cells also received rapamycin therapy (1 mg/kg/day) for 14 days. On day 7 after transplantation, the subsets of peripheral blood T lymphocytes, the pathology of grafts and the infiltration of lymphocytes in grafts were analysed. Untreated gamma-irradiated animals showed a graft median survival time (MST) of 9 days. Animals injected with anergic cells only or receiving rapamycin therapy alone showed MST of 11 and 17 days, respectively. MST of allograft in mice treated with control cells plus rapamycin therapy was 9 days. Animals injected with anergic cells plus rapamycin therapy, but receiving third-party allografts (C57BL/6J), showed an MST of 15 days. However, anergic cell injection plus rapamycin therapy prolonged allograft survival significantly (MST 28 days, P < 0.01). The rejection was mild and tissue architecture was preserved in recipient mice receiving anergic cell injection plus rapamycin therapy. Furthermore, anergic cells and rapamycin therapy decreased the percentage of peripheral blood CD4+ and CD8+ T cells (including CD25+, CD152+, CD154+ and CD28+ subsets) and greatly reduced the infiltrating lymphocytes in allografts (including CD3+, CD4+, CD8+ and CD25+ T cells). In conclusion, the treatment based on anergic cells' adoptive transfer plus rapamycin therapy demonstrated a significant prolongation of murine cardiac allograft survival in a donor antigen-specific manner. This therapeutic protocol alleviated allograft rejection to solid allograft in vivo. PMID- 15787745 TI - Increased frequency of C4B*Q0 alleles in patients with Henoch-Schonlein purpura. AB - Henoch-Schonlein purpura (HSP) is a vasculitis of unknown aetiology, possibly involving immune complexes. The complement system is essential for the clearance of immune complexes. Our aim was to explore the hypothesis that patients with HSP have abnormal complements, contributing to the development of the disease. The study included 56 patients diagnosed with HSP at the Children's Hospital, Iceland between 1984 and 2000, and 98 blood donors as controls. Serum levels of immunoglobulin A, C4A, C4B and mannan-binding lectin were measured and compared between the two groups. C4 null alleles were significantly more common in HSP patients than in controls (P = 0.018) and were carried by 66.1% of the patients compared with 41.2% of the controls. This difference was due to an increased frequency of C4B*Q0 allele in the HSP group (0.25 versus 0.11 in the control group; P = 0.002). The fact that the majority of our patients carried a C4 null allele indicates that children with C4 deficiencies may have an increased risk of developing HSP. This may reflect inadequate complement activity and possibly present an opportunity to identify patients at risk of developing serious morbidity associated with HSP. PMID- 15787746 TI - Quantitative measurement of the levels of melanocortin receptor subtype 1, 2, 3 and 5 and pro-opio-melanocortin peptide gene expression in subsets of human peripheral blood leucocytes. AB - Levels of the melanocortin receptor (MCR) 1, 2, 3 and 5 subtypes and pro-opio melanocortin (POMC) protein mRNA were measured by the real-time quantitative reverse transcriptase polymerase chain reaction method in CD4+ T helper (Th) cells, CD8+ T cytotoxic cells, CD19+ B cells, CD56+ natural killer (NK) cells, CD14+ monocytes and CD15+ granulocytes from healthy donors. We found high levels of all of the MC1, 2, 3 and 5R subtype mRNA in Th cells and moderate levels in NK cells, monocytes and granulocytes. POMC peptide mRNA was found in all examined leucocyte subsets, but only low levels were present in granulocytes. Our findings suggest a co-ordinating role for MCR subtypes and their naturally occurring ligands in the co-operation between innate and adaptive immunity. Moreover, our findings are compatible with earlier finding of MCR-mediated tolerance induction in Th cells. PMID- 15787747 TI - T-cell receptor Vbeta repertoire after myeloablative and reduced intensity conditioning allogeneic haematopoietic stem cell transplantation. AB - T cells play an important role in the adaptive immune system. After haematopoietic stem cell transplantation (HSCT), T-cell function is impaired. This is reflected by the emergence of opportunistic infections, infections that are often difficult to treat because of the patient's insufficient immune function. T-cell receptor reconstitution was studied using CDR3 spectratyping to analyze the diversity of the T-cell repertoire at 3, 6 and 12 months after myeloablative and reduced intensity conditioning (RIC) HSCT in 23 patients. Immune function in vitro was tested by lymphocyte stimulation at 3, 6 and 12 months after HSCT. Lower diversity in the CDR3 repertoire was demonstrated in CD4+ cells after RIC HSCT at 3 and 6 months and in CD8+ cells at 3 months compared with healthy donors. After myeloablative HSCT, lower diversity was seen at 3, 6 and 12 months in CD4+ cells and at 6 and 12 months in CD8+ cells after HSCT. Acute and chronic graft-versus-host-disease (GVHD) did not affect diversity. Responses to phytohaemagglutinin (PHA), Concanavalin A (Con A) and Staphylococcus aureus protein A were significantly lower compared with healthy donors during the first 6 months after RIC HSCT. After myeloablative HSCT, lymphocyte response to Con A was significantly lower at 3 months compared with healthy donors. Decreased responses to cytomegalovirus and varicella zoster virus antigens were seen in patients suffering from acute GVHD grade II or chronic GVHD. The T-cell repertoire is skewed under the first year after HSCT, and immune reconstitution after HSCT with myeloablative and RIC conditioning seems to be comparable. GVHD, infections and age are more important for immune reconstitution than type of conditioning. PMID- 15787753 TI - Quantitative risk assessment of FMD virus transmission via water. AB - Foot-and-mouth disease (FMD) is a viral disease of domesticated and wild cloven hoofed animals. FMD virus is known to spread by direct contact between infected and susceptible animals, by animal products such as meat and milk, by the airborne route, and mechanical transfer on people, wild animals, birds, and by vehicles. During the outbreak of 2001 in the Netherlands, milk from dairy cattle was illegally discharged into the sewerage as a consequence of transport prohibition. This may lead to contaminated discharges of biologically treated and raw sewage in surface water that is given to cattle to drink. The objective of the present study was to assess the probability of infecting dairy cows that were drinking FMD virus contaminated surface water due to illegal discharges of contaminated milk. So, the following data were collected from literature: FMD virus inactivation in aqueous environments, FMD virus concentrations in milk, dilution in sewage water, virus removal by sewage treatment, dilution in surface water, water consumption of cows, size of a herd in a meadow, and dose-response data for ingested FMD virus by cattle. In the case of 1.6 x 10(2) FMD virus per milliliter in milk and discharge of treated sewage in surface water, the probability of infecting a herd of cows was estimated to be 3.3 x 10(-7) to 8.5 x 10(-5), dependent on dilution in the receiving surface water. In the case of discharge of raw sewage, all probabilities of infection were 100 times higher. In the case of little dilution in small rivers, the high level of 8.5 x 10(-3) is reached. For 10(4) times higher FMD virus concentrations in milk, the probabilities of infecting a herd of cows are high in the case of discharge of treated sewage (3.3 x 10(-3) to 5.7 x 10(-1)) and very high in the case of discharge of raw sewage (0.28-1.0). It can be concluded that illegal and uncontrolled discharges of contaminated milk into the sewerage system may lead to high risks to other cattle farms at 6-50 km distance of the location of discharge within one day. This clearly underlines current measures that prohibit such discharges, and also asks for strict control. This risk assessment clearly demonstrated the potential significance of FMD virus transmission via water, and the results will be useful on an international scale, and could also serve as a basis for other FMD risk-assessment models. PMID- 15787751 TI - Comment on "Children's exposure to arsenic from CCA-treated wooden decks and playground structures". PMID- 15787754 TI - Estimation of true Salmonella prevalence jointly in cattle herd and animal populations using Bayesian hierarchical modeling. AB - The Finnish salmonella control program (FSCP) for beef production is based on both randomized and selective testing of herds and animals. Sampling of individual animals in abattoirs is randomized. Herds are selectively tested for salmonella on the basis of clinical symptoms and/or other factors. Risk assessment of FSCP is inherently difficult due to the complexity of the complete data set, especially if the detailed labeling of the testing types is lost. However, for a risk assessment of the whole production chain, methods for exploiting all available data should be considered. For this purpose, a hierarchical Bayesian model of true salmonella prevalence was constructed to combine information at different levels of aggregation: herds in geographical regions and individual animals arriving for slaughter. The conditional (municipality specific) probability of selection of a herd for testing was modeled given the underlying true infection status of the herd and information about the general sampling activity in the specific region. The model also accounted for the overall sensitivity of the sampling methods, both at the herd and at the animal level. In 1999, the 95% posterior probability intervals of true salmonella prevalence in the herd population, in individual cattle, and in slaughter animal populations were [0.54%, 1.4%] (mode 0.8%), [0.15%, 0.39%] (mode 0.2%), and [0.12%, 0.36%] (mode 0.2%), respectively. The results will be further exploited in other risk assessments focusing on the subsequent parts of the beef production chain, and in evaluation of the FSCP. PMID- 15787755 TI - Uncertainty distribution associated with estimating a proportion in microbial risk assessment. AB - The uncertainty associated with estimates should be taken into account in quantitative risk assessment. Each input's uncertainty can be characterized through a probabilistic distribution for use under Monte Carlo simulations. In this study, the sampling uncertainty associated with estimating a low proportion on the basis of a small sample size was considered. A common application in microbial risk assessment is the estimation of a prevalence, proportion of contaminated food products, on the basis of few tested units. Three Bayesian approaches (based on beta(0, 0), beta(1/2, 1/2), and beta(l, 1)) and one frequentist approach (based on the frequentist confidence distribution) were compared and evaluated on the basis of simulations. For small samples, we demonstrated some differences between the four tested methods. We concluded that the better method depends on the true proportion of contaminated products, which is by definition unknown in common practice. When no prior information is available, we recommend the beta (1/2, 1/2) prior or the confidence distribution. To illustrate the importance of these differences, the four methods were used in an applied example. We performed two-dimensional Monte Carlo simulations to estimate the proportion of cold smoked salmon packs contaminated by Listeria monocytogenes, one dimension representing within-factory uncertainty, modeled by each of the four studied methods, and the other dimension representing variability between companies. PMID- 15787756 TI - A model of hygiene practices and consumption patterns in the consumer phase. AB - A mathematical model is presented, which addresses individual hygiene practices during food preparation and consumption patterns in private homes. Further, the model links food preparers and consumers based on their relationship to household types. For different age and gender groups, the model estimates (i) the probability of ingesting a meal where precautions have not been taken to avoid the transfer of microorganisms from raw food to final meal (a risk meal), exemplified by the event that the cutting board was not washed during food preparation, and (ii) the probability of ingesting a risk meal in a private home, where chicken was the prepared food item (a chicken risk meal). Chicken was included in the model, as chickens are believed to be the major source of human exposure to the foodborne pathogen Campylobacter. Monte Carlo simulations showed that the probability of ingesting a risk meal was highest for young males (aged 18-29 years) and lowest for the elderly above 60 years of age. Children aged 0-4 years had a higher probability of ingesting a risk meal than children aged 5-17 years. This difference between age and gender groups was ascribed to the variations in the hygiene levels of food preparers. By including the probability of ingesting a chicken meal at home, simulations revealed that all age groups, except the group above 60 years of age, had approximately the same probability of ingesting a chicken risk meal, the probability of females being slightly higher than that of males. The simulated results show that the probability of ingesting a chicken risk meal at home does not only depend on the hygiene practices of the persons preparing the food, but also on the consumption patterns of consumers, and the relationship between people preparing and ingesting food. This finding supports the need of including information on consumer behavior and preparation hygiene in the consumer phase of exposure assessments. PMID- 15787757 TI - Modeling growth of Clostridium perfringens in pea soup during cooling. AB - Clostridium perfringens is a pathogen that mainly causes food poisoning outbreaks when large quantities of food are prepared. Therefore, a model was developed to predict the effect of different cooling procedures on the growth of this pathogen during cooling of food: Dutch pea soup. First, a growth rate model based on interpretable parameters was used to predict growth during linear cooling of pea soup. Second, a temperature model for cooling pea soup was constructed by fitting the model to experimental data published earlier. This cooling model was used to estimate the effect of various cooling environments on average cooling times, taking into account the effect of stirring and product volume. The growth model systematically overestimated growth of C. perfringens during cooling in air, but this effect was limited to less than 0.5 log N/ml and this was considered to be acceptable for practical purposes. It was demonstrated that the growth model for C. perfringens combined with the cooling model for pea soup could be used to sufficiently predict growth of C. perfringens in different volume sizes of pea soup during cooling in air as well as the effect of stirring, different cooling temperatures, and various cooling environments on the growth of C. perfringens in pea soup. Although fine-tuning may be needed to eliminate inaccuracies, it was concluded that the combined model could be a useful tool for designing good manufacturing practices (GMP) procedures. PMID- 15787759 TI - A poultry-processing model for quantitative microbiological risk assessment. AB - A poultry-processing model for a quantitative microbiological risk assessment (QMRA) of campylobacter is presented, which can also be applied to other QMRAs involving poultry processing. The same basic model is applied in each consecutive stage of industrial processing. It describes the effects of inactivation and removal of the bacteria, and the dynamics of cross-contamination in terms of the transfer of campylobacter from the intestines to the carcass surface and the environment, from the carcasses to the environment, and from the environment to the carcasses. From the model it can be derived that, in general, the effect of inactivation and removal is dominant for those carcasses with high initial bacterial loads, and cross-contamination is dominant for those with low initial levels. In other QMRA poultry-processing models, the input-output relationship between the numbers of bacteria on the carcasses is usually assumed to be linear on a logarithmic scale. By including some basic mechanistics, it is shown that this may not be realistic. As nonlinear behavior may affect the predicted effects of risk mitigations; this finding is relevant for risk management. Good knowledge of the variability of bacterial loads on poultry entering the process is important. The common practice in microbiology to only present geometric mean of bacterial counts is insufficient: arithmetic mean are more suitable, in particular, to describe the effect of cross-contamination. The effects of logistic slaughter (scheduled processing) as a risk mitigation strategy are predicted to be small. Some additional complications in applying microbiological data obtained in processing plants are discussed. PMID- 15787758 TI - Meta-analysis of food safety information based on a combination of a relational database and a predictive modeling tool. AB - The management of microbial risk in food products requires the ability to predict growth kinetics of pathogenic microorganisms in the event of contamination and growth initiation. Useful data for assessing these issues may be found in the literature or from experimental results. However, the large number and variety of data make further development difficult. Statistical techniques, such as meta analysis, are then useful to realize synthesis of a set of distinct but similar experiences. Moreover, predictive modeling tools can be employed to complete the analysis and help the food safety manager to interpret the data. In this article, a protocol to perform a meta-analysis of the outcome of a relational database, associated with quantitative microbiology models, is presented. The methodology is illustrated with the effect of temperature on pathogenic Escherichia coli and Listeria monocytogenes, growing in culture medium, beef meat, and milk products. Using a database and predictive models, simulations of growth in a given product subjected to various temperature scenarios can be produced. It is then possible to compare food products for a given microorganism, according to its growth ability in these products, and to compare the behavior of bacteria in a given foodstuff. These results can assist decisions for a variety of questions on food safety. PMID- 15787760 TI - A linear model for managing the risk of antimicrobial resistance originating in food animals. AB - A linear population risk model used by the U.S. Food and Drug Administration (FDA) Center for Veterinary Medicine (CVM) estimates the risk of human cases of campylobacteriosis caused by fluoroquinolone-resistant Campylobacter. Among the cases of campylobacteriosis attributed to domestically produced chicken, the fluoroquinolone resistance is assumed to result from the use of fluoroquinolones in poultry in the United States. Properties of the linear population risk model are contrasted with those of a farm-to-fork model commonly used for microbial risk assessments. The utility of the linear population model for the purpose for which it was used by CVM is discussed. PMID- 15787761 TI - A structured expert judgment study for a model of Campylobacter transmission during broiler-chicken processing. AB - A structured expert judgment study was organized to obtain input data for a microbial risk-assessment model describing the transmission of campylobacter during broiler-chicken processing in the Netherlands. More specially, the expert study was aimed at quantifying the uncertainty on input parameters of this model and focused on the contamination of broiler-chicken carcasses with campylobacter during processing. Following the protocol for structured expert judgment studies, expert assessments were elicited individually through subjective probability distribution functions. The classical model was used to aggregate the individual experts' distributions in order to obtain a single combined distribution per variable. Three different weighting schemes were applied, including equal weighting and performance-based weighting with and without optimalization of the combined distributions. The individual experts' weights were based on their performance on the seed variables. Results of the various weighting schemes are presented in terms of performance, robustness, and combined distributions of the seed variables and some of the query variables. All three weighting schemes had adequate performance, with the optimized combined distributions significantly outperforming both the equal weight and the nonoptimized combined distributions. Hence, this weighting scheme, having adequate robustness, was chosen for further processing of the results. PMID- 15787763 TI - Proportional hazards in information security. AB - Nonparametric methods can be used to analyze failure times and estimate probability distributions for failures of systems due to successful attacks on confidentiality, integrity, and availability in information security. However, such methods do not take full advantage of supplemental information regarding the configurations of systems in an information infrastructure that is usually also available. One approach, which does take advantage of such information, views the risks of systems failing from various causes as competing risks and determines the correlation coefficients of different treatments to system longevity. Since the times and causes of failure in such studies are usually uncorrelated, the hazards associated with each risk are proportional. By correlating system survival times to the use of specific design enhancements and security countermeasures, as well as to system exposure based on choice of operational functionality, guidance can be obtained for making investments in information security. PMID- 15787762 TI - Logic and motivation in risk research: a nuclear waste test case. AB - After two decades of massive investigation, federal approval of a nuclear waste site is drawing to a close. Large-scale research to assure that major hazards such as this one are socially acceptable is often highly inefficient, as here. A regulatory remedy would be to require not only that risk currently assessed be acceptable, but also that risk would remain acceptable given any new information. Research to test compliance with these rules has to be cost effective. Research activities should be managed according to an explicit discipline that can be imposed on powerful conflicting interests. They might be required to (1) set targets for the first- and second-order risk assessments, (2) allocate research resources to close any gap between current and target assessments cost effectively, and (3) reallocate resources, as evolving findings dictate. The interests of license applicant (Department of Energy, DOE) and society are distinguished: the former would want the application approved; the latter would want to reject an unacceptable facility. PMID- 15787764 TI - Health effects caused by primary fine particulate matter (PM2.5) emitted from buses in the Helsinki metropolitan area, Finland. AB - Fine particle (PM(2.5)) emissions from traffic have been associated with premature mortality. The current work compares PM(2.5)-induced mortality in alternative public bus transportation strategies as being considered by the Helsinki Metropolitan Area Council, Finland. The current bus fleet and transportation volume is compared to four alternative hypothetical bus fleet strategies for the year 2020: (1) the current bus fleet for 2020 traffic volume, (2) modern diesel buses without particle traps, (3) diesel buses with particle traps, and (4) buses using natural gas engines. The average population PM(2.5) exposure level attributable to the bus emissions was determined for the 1996-1997 situation using PM(2.5) exposure measurements including elemental composition from the EXPOLIS-Helsinki study and similar element-based source apportionment of ambient PM(2.5) concentrations observed in the ULTRA study. Average population exposure to particles originating from the bus traffic in the year 2020 is assumed to be proportional to the bus emissions in each strategy. Associated mortality was calculated using dose-response relationships from two large cohort studies on PM(2.5) mortality from the United States. Estimated number of deaths per year (90% confidence intervals in parenthesis) associated with primary PM(2.5) emissions from buses in Helsinki Metropolitan Area in 2020 were 18 (0 55), 9 (0-27), 4 (0-14), and 3 (0-8) for the strategies 1-4, respectively. The relative differences in the associated mortalities for the alternative strategies are substantial, but the number of deaths in the lowest alternative, the gas buses, is only marginally lower than what would be achieved by diesel engines equipped with particle trap technology. The dose-response relationship and the emission factors were identified as the main sources of uncertainty in the model. PMID- 15787765 TI - Risk/benefit assessments of human diseases: optimum dose for intervention. AB - A simple procedure is proposed in order to quantify the tradeoff between a loss suffered from an illness due to exposure to a microbial pathogen and a loss due to a toxic effect, perhaps a different illness, induced by a disinfectant employed to reduce the microbial exposure. Estimates of these two types of risk as a function of disinfectant dose and their associated relative losses provide information for the estimation of the optimum dose of disinfectant that minimizes the total expected loss. The estimates of the optimum dose and expected relative total loss were similar regardless of whether the beta-Poisson, log-logistic, or extreme value function was used to model the risk of illness due to exposure to a microbial pathogen. This is because the optimum dose of the disinfectant and resultant expected minimum loss depend upon the estimated slope (first derivative) of the models at low levels of risk, which appear to be similar for these three models at low levels of risk. Similarly, the choice among these three models does not appear critical for estimating the slope at low levels of risk for the toxic effect induced by the use of a disinfectant. For the proposed procedure to estimate the optimum disinfectant dose, it is not necessary to have absolute values for the losses due to microbial-induced or disinfectant-induced illness, but only relative losses are required. All aspects of the problem are amenable to sensitivity analyses. The issue of risk/benefit tradeoffs, more appropriately called risk/risk tradeoffs, does not appear to be an insurmountable problem. PMID- 15787766 TI - Issues related to time averaging of exposure in modeling risks associated with intermittent exposures to lead. AB - Typical exposures to lead often involve a mix of long-term exposures to relatively constant exposure levels (e.g., residential yard soil and indoor dust) and highly intermittent exposures at other locations (e.g., seasonal recreational visits to a park). These types of exposures can be expected to result in blood lead concentrations that vary on a temporal scale with the intermittent exposure pattern. Prediction of short-term (or seasonal) blood lead concentrations arising from highly variable intermittent exposures requires a model that can reliably simulate lead exposures and biokinetics on a temporal scale that matches that of the exposure events of interest. If exposure model averaging times (EMATs) of the model exceed the shortest exposure duration that characterizes the intermittent exposure, uncertainties will be introduced into risk estimates because the exposure concentration used as input to the model must be time averaged to account for the intermittent nature of the exposure. We have used simulation as a means of determining the potential magnitude of these uncertainties. Simulations using models having various EMATs have allowed exploration of the strengths and weaknesses of various approaches to time averaging of exposures and impact on risk estimates associated with intermittent exposures to lead in soil. The International Commission of Radiological Protection (ICRP) model of lead pharmacokinetics in humans simulates lead intakes that can vary in intensity over time spans as small as one day, allowing for the simulation of intermittent exposures to lead as a series of discrete daily exposure events. The ICRP model was used to compare the outcomes (blood lead concentration) of various time averaging adjustments for approximating the time-averaged intake of lead associated with various intermittent exposure patterns. Results of these analyses suggest that standard approaches to time averaging (e.g., U.S. EPA) that estimate the long-term daily exposure concentration can, in some cases, result in substantial underprediction of short-term variations in blood lead concentrations when used in models that operate with EMATs exceeding the shortest exposure duration that characterizes the intermittent exposure. Alternative time-averaging approaches recommended for use in lead risk assessment more reliably predict short-term periodic (e.g., seasonal) elevations in blood lead concentration that might result from intermittent exposures. In general, risk estimates will be improved by simulation on shorter time scales that more closely approximate the actual temporal dynamics of the exposure. PMID- 15787767 TI - Research strategies for magnetic fields and cancer. AB - Widespread concerns about whether electric and magnetic fields (EMF) could adversely affect human health have been raised in epidemiologic studies reported since the 1980s. Possible EMF health effects have been widely publicized in the popular press since that time. We consider here three possible mechanisms of action of EMF on childhood leukemia. We identify the first as "magnetic fields": this hypothesis relates the average level of magnetic field to the incidence of childhood leukemia. We identify a second, recently proposed, mechanism as "contact current": this hypothesis relates the low voltage and consequent current that occurs on the domestic water pipe, due to U.S. grounding practices, as a source for exposure of children. The third hypothesis is that the relationship observed is spurious. Using a modified example taken from the work of Von Winterfeldt and Keeney, we use Decision Analysis to estimate the value of information for distinguishing between the three hypotheses. We believe that this improves on the usual process for deciding on research budgets. Depending on which hypothesis we favor a priori, the value of being informed ranges from US 101 dollars to US 233 dollars per "problem household." Since there could be as many as 2 million such households, the value of information for resolving this issue could approach half a billion dollars! We find that there is no value of information for finding the odds ratio given the contact current hypothesis. In writing this article, we have consciously kept the computations as simple as possible so as to engage the reader's attention and interest. In a penultimate section, we suggest numerous possible extensions for a group interested in discussing and deciding on the value of research on the relationship between magnetic fields and cancer. PMID- 15787768 TI - Integration of stochastic effects and data uncertainties into the design of process equipment. AB - Stochastic effects and data uncertainties are present in any engineering calculation. Their impact may be particularly important if they concern the design of process equipment. A calculation model for the dynamic behavior of a heat exchanger and procedures to deal with the related uncertainties are presented. Their propagation through the calculation by means of a Monte Carlo approach is shown. The temperature at the heat exchanger outlet and the step response of a sudden variation in the heat exchanger inlet temperature are simulated and evaluated by way of example. It is demonstrated that the inclusion of stochastic effects and uncertainties provides a more reliable basis for design decisions and hence reduces the probability of errors. PMID- 15787769 TI - Trust in risk regulation: cause or consequence of the acceptability of GM food? AB - Although there is ample empirical evidence that trust in risk regulation is strongly related to the perception and acceptability of risk, it is less clear what the direction of this relationship is. This article explores the nature of the relationship, using three separate data sets on perceptions of genetically modified (GM) food among the British public. The article has two discrete but closely interrelated objectives. First, it compares two models of trust. More specifically, it investigates whether trust is the cause (causal chain account) or the consequence (associationist view) of the acceptability of GM food. Second, this study explores whether the affect heuristic can be applied to a wider number of risk-relevant concepts than just perceived risk and benefit. The results suggest that, rather than a determinant, trust is an expression or indicator of the acceptability of GM food. In addition, and as predicted, "affect" accounts for a large portion of the variance between perceived risk, perceived benefit, trust in risk regulation, and acceptability. Overall, the results support the associationist view that specific risk judgments are driven by more general evaluative judgments The implications of these results for risk communication and policy are discussed. PMID- 15787770 TI - A new look at the psychometric paradigm of perception of hazards. AB - The psychometric paradigm has been the most influential model in the field of risk analysis. The "cognitive maps" of hazards produced by the paradigm seem to explain how laypeople perceive the various risks they face. Because most of the studies used aggregated data, analyzed using principal component analysis, it is not known whether the model neglects individual differences in risk perception. There has been much criticism on the fact that few studies have examined individual differences in the cognitive representation of hazards. In order to detect and describe the internal structure of the three-way data, we conducted a three-way component analysis (3MPCA). Data for the present analysis were derived from a mail survey conducted in Switzerland. Participants were asked to judge 9 attributes for 26 hazards. Individual differences in the cognitive representation of hazards were correlated with external variables (e.g., general trust). The results suggest that methods permitting individual differences should be used more frequently and that utilizing different methods could provide greater insight into the cognitive representation of risks. PMID- 15787775 TI - Abortion discourse in Bolivian hospital contexts: doctors' repertoire conflicts and the Saving Women device. AB - The author proposes going Beyond attitudes (Potter and Wetherell 1987) to a more nuanced assessment of doctors' discursive variations. Through an application of Gilbert and Mulkay's (1984) interpretative repertoires, she defined three voices- technical, normative and pragmatic--in which Bolivian doctors spoke of abortion. In State and social security hospital contexts, doctors hastened to express compliance with government policy and institutional norms regulating abortion and postabortion care. Technical and pragmatic considerations, however, often entered into conflict with established rules. When contradictions became apparent in their own discourse, doctors regularly drew on the Saving Women device. This accounting strategy enabled them to save face as up-to-date professionals through justifying temporary deviance from norms in terms of benefit to women treated. The author describes her development of the repertoires, their validation with different medical audiences, and doctors' critical appropriation of the model to explain their own discursive variations. PMID- 15787774 TI - The personal significance of home: habitus and the experience of receiving long term home care. AB - The physical, symbolic and experiential aspects of receiving long-term care are examined in this paper using Bourdieu's concepts of habitus and field. We draw on data from an ethnographic study of home care in 16 homes in urban, rural and remote locations in Ontario, Canada. Across all cases, data about domestic and caregiving routines were gathered through observation, interviews with clients and/or the primary family caregiver, interviews with service providers and videotaped tours of the home. Based on the analysis of these data, we argue that a transposition of logics and practices occurred when the domestic and health care fields were superimposed within the spaces of the home. Although all of the care recipients and their family caregivers indicated a strong preference for home care over institutional care, their experiences and practices within their homes were disrupted and reconfigured by the insertion of logics emanating from the healthcare field. These changes were manifested in three main themes: the politics of aesthetics; the maintenance of order and cleanliness; and transcending the limitations of the home. In each of these dimensions, it became apparent that care recipients engaged in improvisatory social practices that reflected their ambiguous and changing habitus or social location. The material spaces of their homes signified, or prompted, altered or changing social placement. PMID- 15787776 TI - Explaining caesarean section in Salvador da Bahia, Brazil. AB - In Salvador da Bahia the caesarean section rate is excessive, as it is in Brazil as a whole. It is the standard form of delivery in private hospitals, though vaginal delivery still predominates in the public sector. This paper investigates the social context of these styles of childbirth, arguing that the connections between both sectors sustain this situation. Exploring the factors leading to the preference in private and insurance-funded maternity wards, it examines critically two diverging positions on the cause of the rate: that women's cultural preferences for abdominal birth lie behind it; or that obstetricians' self-interest is to blame. The paper critiques the theory of culture behind the first stance and questions the theoretical weight placed on individual action in the second. It argues that no particular social group is the principal cause of the excessive use of caesarean section to deliver babies. Rather, a host of factors converge in sustaining this practice. Finally, the paper stresses that the system as a whole, not any particular group, must be changed if the rate is to be lowered significantly. For this, political will is required. PMID- 15787777 TI - Birth and social class: Northern Thai women's lived experiences of caesarean and vaginal birth. AB - This paper, based on in-depth interviews with Thai women in Northern Thailand, contributes to a sociological understanding of women's childbirth discourses. The findings indicate that the lived experiences of birth differ between individual women. It clearly shows that social resources such as financial resources and education play a salient role in shaping the embodied experience of birth among women in Northern Thailand. Because of their 'everyday lifestyle', middle class women have more control over the experience of childbirth than that of the rural poor women. Middle class women are able to choose where to give birth, have access to private care and actively seek medical technology as a way to have control over their birth. Their material resources enable their choices. These choices seem to be denied to the rural poor women. But not all rural poor women are passive victims of their material resources. No matter how limited the resources women have, they use them. Hence, there are some poor women who actively seek birthing care that enables them to have more control. But regardless of their social positions (urban middle class or rural poor), obstetric interventions are commonly experienced, and most women perceive caesarean birth in a positive light. Several discourses are employed to explain these findings including women's interpretations of their lived world including risk and the medicalisation of childbirth in Thailand. Taking a feminist standpoint, I argue that differences between women need to be taken into account in providing care to women in childbirth so that sensitive and appropriate birthing care can be achieved. PMID- 15787778 TI - Securing our genetic health: engendering trust in UK Biobank. AB - The recent development of genetic databases, or 'biobanks', in a number of countries reflects scientists' and policy makers' beliefs in the future health benefits to be derived from genetics research. In Britain, however, a proposal for a genetic database, UK Biobank, has been the focus of a number of concerns. Establishing consent and legitimacy for any controversial biomedical research involving the participation of human subjects is difficult; it is however, acute for UK Biobank given the scale of the project and the criticisms levelled at it. Analysing recently published documents pertaining to UK Biobank, this article examines how consent for the project has been discursively framed and how this is reflected in its governance. It is argued that the problem of organising consent has been framed narrowly in terms of adherence to a well-established repertoire of institutional mechanisms which serves to limit debate on the substantive issues at stake. There is little evidence of reflection on the adequacy of such mechanisms for dealing with the unique challenges posed by UK Biobank, including achieving the confidence and participation of a population with diverse perspectives on genetic research. It is concluded that a restricted public discourse about UK Biobank may contribute to a decline in confidence in regulatory systems governing biotechnology and science more generally. PMID- 15787780 TI - Tacrolimus pharmacokinetics in pediatric renal transplant patients. PMID- 15787781 TI - Health-related quality of life after pediatric heart or heart-lung transplantation: Where do we go from here? PMID- 15787782 TI - New kids on the block: an old problem for a growing pediatric population. PMID- 15787783 TI - Indications for varicella vaccination post-transplant. PMID- 15787784 TI - From whence does biliary atresia arise? PMID- 15787785 TI - Current status of ABO-incompatible kidney transplantation in children. AB - This article reviews the current status of ABO-incompatible kidney transplantation in the pediatric population. ABO blood type incompatibility between a donor and recipient was generally considered a contraindication to kidney transplantation because of the associated high risk for hyperacute rejection. However, due to a severe shortage of suitable cadaveric allografts, much effort has been made over the last decade to investigate whether successful and effective kidney transplantation is possible across the ABO blood group barrier. At present, ABO-incompatible kidney transplantation has been shown to be a valid alternative even for children with end stage renal disease. In this review, we will discuss protocols available for successfully performing ABO incompatible kidney transplantation in children: (1) pre-transplant extracorporeal immunomodulation with removal of pre-existing anti-A and/or anti-B antibodies; (2) immunosuppressive therapy and anti-rejection therapy; (3) splenectomy and the associated infectious complication in asplenic children. Also, we will speculate regarding the mechanisms underlying accommodation following transplantation. PMID- 15787786 TI - Post-transplant therapy with high-dose intravenous gammaglobulin: Applications to treatment of antibody-mediated rejection. AB - B-cells and their products (antibodies) are now recognized as important mediators of allograft rejection. This represents a significant departure from previous doctrine where the T-cells were felt to be of paramount importance. Antibody mediated rejection (AMR) has emerged as a significant and common complication of transplantation. The development of donor-specific (anti-HLA class I and class II) antibodies is known to correlate strongly with the development of AMR. Recognition of the unique features of AMR that were often confused with non specific acute tubular injury is aided considerably by improvements in monitoring of anti-HLA antibodies and the immunopathologic demonstration of C4d staining in affected capillary beds. Although imperfect, the demonstration of C4d (a complement breakdown product) staining in an allograft, especially accompanied by the presence of anti-HLA antibodies in the recipient sera, strongly suggests a diagnosis of AMR. Thus, AMR is a complement-dependent, antibody-mediated disorder. AMR can occur as a de novo complication of transplantation in individuals not previously recognized to be sensitized to HLA antigens, but more often occurs as a complication of desensitization therapies in highly-HLA sensitized patients. AMR may also constitute a significant portion of what is now referred to as chronic allograft nephropathy (CAN). The prognosis of C4d (+) AMR is poor as current therapies for treatment of AR are directed primarily at the T cell. Until recently, no therapeutic options existed to address this problem from a primary etiological standpoint. Here we discuss the use of high dose IVIG as an option for treatment of AMR. We have significant experience with this approach which is outlined here. IVIG has many ideal advantages as a therapy for AMR. First, it can down regulate B-cell activation and antibody production, second, it can induce anti-inflammatory cytokines and contains blocking antiidiotyic antibodies to anti-HLA antibodies and third, IVIG has the unique ability to block complement-mediated injury through inhibition of C3 activation. Further clinical trials are necessary to prove efficacy for treatment of AMR. PMID- 15787787 TI - Effect of age, ethnicity, and glucocorticoid use on tacrolimus pharmacokinetics in pediatric renal transplant patients. AB - Tacrolimus has become an effective alternative to cyclosporine as a component of primary immunosuppression in pediatric renal transplant patients, but the information on the pharmacokinetic characteristics of tacrolimus in young patients is still limited. The primary objective of this study was to determine the effect of patient age, ethnicity, and concurrent steroid administration on tacrolimus pharmacokinetics in pediatric renal transplant patients. The study population consisted of 30 pediatric patients, age 1.5-18.6 yr, who received a kidney transplant between July 1999 and February 2004. After twice daily dosing was stabilized based on clinical judgment, at least 5 days postoperatively, tacrolimus levels were drawn prior to, and 1, 2, 4, 8, and 12 h after the morning dose. The mean dose of tacrolimus was 0.12 mg/kg/dose. Mean trough level was 11.9 +/- 5.0 ng/mL. Mean area under the curve (AUC) was 192 +/- 84 with a range of 78 360 h x (ng/mL). The correlation between trough level and AUC was only fair (r = 0.74); later time points correlated better with AUC, and an excellent correlation (r = 0.96) was obtained between the mean of trough and 2-h level (C(2)) and AUC. There was a negative correlation between age and dose per body weight (r = 0.68). African-American patients had marginally lower drug exposure with similar dosing. Three age groups (<5, 5-12, and >12 yr) were compared with respect to dosing and AUC. Despite similar AUC in all three groups, the mean dose per kg required to achieve the AUC was 2.7- and 1.9-fold higher in the <5 and 5-12-yr groups, respectively, compared with the >12-yr group. Nine of the 30 patients were on a totally steroid-free regimen. Their tacrolimus dose and trough levels were similar to those of steroid-exposed patients, but their mean AUC was 41% higher. Our results show an inverse correlation between age and required tacrolimus dose, wide interindividual variation, and greater exposure with steroid-free regimen despite no change in trough level. PMID- 15787788 TI - Beyond the first year after pediatric heart or heart-lung transplantation: Changes in cognitive function and behaviour. AB - With the increasing use and improved survival rates of heart and lung transplantation as treatments for children with end-stage heart or lung disease, attention is focusing on the longer term psychological implications of these procedures. This paper focuses on the changes in cognitive development and behaviour in a group of 47 children who were seen 12 months and 2 yr after transplantation. There were 24 boys and 23 girls, mean age at transplantation was 8.3 yr (s.d. 5.3 yr), with a range of 0.3-15.1 yr. Assessments were made of developmental level, cognitive ability and problem behaviours, using previously validated measures, and comparisons were made with physically healthy children. For children under three and a half years of age there was a decrease over time in scores on all developmental parameters, with the change reaching significance on the scale assessing eye-hand coordination and on the overall IQ. Whilst all scores were within the normal range, they were at a significantly lower level than those of the healthy children. In contrast, there were no changes over time on any measures of cognitive or academic ability for older children, with correlations between 12 month and 2 yr scores being highly significant. The rate of behaviour problems at home at 12 months was 22%, compared with 34% at 2 yr post-transplant, which was higher than that found in the healthy children. Conversely, there was a drop in the prevalence of behaviour problems at school from 23% at 12 months to 9% at 2 yr. It is concluded that a significant minority of children and adolescents experience psychological difficulties 2 yr after transplant, with particular areas of concern focusing on development in the younger children and the occurrence of behaviour problems at home across the age range. PMID- 15787789 TI - Improved long-term allograft function in pediatric renal transplantation with mycophenolate mofetil. AB - MMF has been shown to decrease the incidence of acute rejection in children and adults at 1 and 3 yr. Other beneficial effects of MMF have been more difficult to demonstrate. Our open-labeled study presents a 5-yr data for patients and graft survival, allograft function, and growth in MMF-treated patients. The trial included 29 patients who were treated with MMF in combination with cyclosporine and methylprednisone. Patients were compared with a preceding group of 29 patients treated with AZA instead of MMF. Patient and graft survival rate 5 yr after transplantation were 97 and 90% in the MMF group vs. 93 and 83% in the AZA group (p: NS). Acute rejection was 20.6% in the MMF group vs. 58.6% in the AZA group (p < 0.01). Chronic rejection was 10.3% in the MMF group and 25% in the AZA group (p: NS). The changes in the creatinine clearance from baseline to 5 yr (Delta) were different between groups (-6.0 +/- 5.1 mL/min/1.73 m(2) in the MMF group vs. -22.2 +/- 7.6 mL/min/1.73 m(2) in the AZA group, p < 0.05). Also, the slope of 1/Scr showed a significant lower incidence of worsening renal function after the second year of renal transplantation (p < 0.0001) in the MMF group compared with the AZA group. Delta Height SDS in prepubertal patients was 0.3 +/- 0.4 SDS in the MMF group vs. -0.8 +/- 0.2 SDS in the AZA group (p < 0.05). This study shows that long-term MMF therapy has resulted in a decrease in acute rejection and was associated with a protection against renal function deterioration. The use of MMF enables a reduction in the dose of steroids and leads to a linear growth improvement of children after renal transplantation. PMID- 15787790 TI - Invasive pneumococcal disease in pediatric organ transplant recipients: a high risk population. AB - There are few studies on invasive pneumococcal disease in pediatric transplant recipients. Given this fact plus the advent of pneumococcal conjugate vaccines, we conducted a retrospective study at a major pediatric transplant center. The objectives were to determine the incidence and outcomes of invasive pneumococcal diseases in the patient population and to examine the timing of these infections after transplantation. We determined that invasive disease occurred at a rate that was significantly greater than the rate extrapolated from generally healthy children <5 yr of age (176 episodes per 100 000 children per year vs. 35-68.3 per 100 000 children per year). In addition, disease occurred at a median of approximately 20 months after transplantation, thereby theoretically allowing enough time for vaccination with the 7-valent conjugate vaccine. The study also documented significant missed vaccination opportunities. PMID- 15787791 TI - Sclerosing peritonitis after intestinal transplantation in children. AB - Long-term graft dysfunction and/or graft loss after intestinal transplantation (ITx) is a significant concern. Sclerosing peritonitis (SP) is a manifestation of chronic allograft failure and its presence may also include classic arterial obliterative arteriopathy (OA) as in chronic rejection. We describe the clinical presentation and management of SP occurring after ITx in children. Case records of 121 children undergoing ITx from 1990 to 2003 were reviewed. Three children (2.4%) presented with SP of the intestine allograft at a mean time of 6.6 yr following ITx as follows: age at Tx (yr) 8.2, and 3.7, with indication for ITx being gastroschisis in two and midgut volvulus in one patient. Type of ITx was isolated intestine in one and liver/intestine in two patients. Gross findings of SP included fibrosis/strictures; microscopically SP showed fibrosis/serositis, and fibrous adhesions; one patient had evidence of chronic allograft vasculopathy. All patients presented with clinical signs and symptoms of bowel obstruction and gastrointestinal contrast studies confirmed distal ileal obstruction (DIO). Operative findings confirmed SP and DIO in all patients; all patients were initially treated with distal segmental intestine allograft resection and lysis of the fibrous peel. All three patients recovered, although two required repeat laparotomy, there is only one long-term survival. SP after ITx may be a different manifestation of long-term intestine allograft degeneration. Surgical resection appears to offer palliation. PMID- 15787792 TI - Seroprevalence of antibodies against varicella-zoster virus and response to the varicella vaccine in pediatric renal transplant patients. AB - The severity of varicella-zoster virus (VZV) in immunocompromised children, especially in those receiving renal transplants, is well known. However, the use of live attenuated virus vaccine in this population is controversial. This study aimed to: (i) assess the immunization status of pediatric renal transplant recipients at our center; (ii) determine the anti-VZV antibody titers in such patients; (iii) evaluate the response to VZV vaccine in seronegative children and in those who present low antibody titers (defined as <500 mAU/mL). Vaccinated children were monitored for adverse effects for 8 wk after vaccination. Fifty patients with a mean age of 13.7 yr (range, 3-17 yr) were enrolled. In 49, blood samples were collected and antibodies were screened using ELISA. Seropositivity to VZV was found in 43 (88%), and antibody titers were >/=500 mAU/mL in 37 (75.5%). Of the 12 children who were eligible for vaccination and had antibody titers <500 mAU/mL, one developed varicella before vaccination, two did not meet the inclusion criteria, and three parents refused the vaccination. In the six vaccinated children, there were no adverse reactions to the vaccine, and four (66.6%) responded with anti-VZV titers >/=500 mAU/mL 6-8 wk after vaccination. In conclusion, after renal transplantation, varicella vaccine is safe with a 66% rate of conversion to high antibody titers. PMID- 15787793 TI - Open lung biopsy in pediatric patients with respiratory failure after abdominal transplantation. AB - To understand the utility of open lung biopsy (OLB) in the evaluation of respiratory failure in pediatric abdominal organ transplant we reviewed the records of nine children in this patient population who underwent an OLB. Eight of nine patients had undergone a previous non-diagnostic bronchoalveolar lavage. Biopsies were performed at the bedside in the pediatric intensive care unit and tissue was processed by the Department of Pathology with special stains for infectious agents. There were no significant complications of OLB. A specific treatable etiology was identified in four patients (respiratory syncytial virus, adenovirus, graft-vs.-host disease and post-transplant lymphoproliferative disease), leading to a change in therapy and survival in two. Overall survival was 44%. Given the low morbidity, OLB as performed in this study appears appropriate in this patient population. PMID- 15787794 TI - Renal function outcome in pediatric liver transplant recipients. AB - The orthotopic liver transplantation (OLT) allows survival of children followed for severe hepatic injury, provided that the immunosuppressive treatment is prolonged. The nephrotoxicity of cyclosporine predicts the long-term outcome of the adult patients receiving a liver transplant. The aim of this study was to determine the long-term outcome of renal function in children receiving OLT. This study included 12 children, with a median for age of 7.1 yr (2-15 yr) at the time of OLT. The duration of follow-up was at least 4 yr, being 7 yr in 10 patients and more than 10 yr in seven. Renal function was evaluated with the serum level of creatinine, calculated glomerular filtration rate (cGFR), and measurement of glomerular filtration rate using chrome 51 ethylenediaminetetraacetate ((51)Cr EDTA) clearance performed at least once during follow-up. The doses and the serum concentrations (C(0)) of cyclosporine were reported at each study time. The cGFR decreased significantly 2 yr after the OLT [median (range): 106 mL/min/1.73 m(2) (71-150) at the time of OLT vs. 85 mL/min/1.73 m(2) (57-128) 2 yr after the OLT, p = 0.03], and decreased again between 7 and 10 yr after OLT [median (range): 99 mL/min/1.73 m(2) (76-125) 7 yr after OLT vs. 81 mL/min/1.73 m(2) (66-140) 10 yr after OLT, p = 0.04]. Six patients developed chronic renal failure (cGFR from 57 to 80 mL/min/1.73 m(2)) 2 yr after OLT associated with high doses of cyclosporine [median (range): 8.8 mg/kg/day (3.5-13)]. The cGFR overestimated renal function by 16% compared with the isotopic measurement of GFR (p = 0.03). Using the (51)Cr EDTA measurement, six of seven patients followed up more than 10 yr after OLT presented mild (n = 3) or moderate (n = 3) chronic renal failure. In our study, the majority of OLT recipients developed a chronic renal failure 10 yr after transplantation. Cyclosporine seems to be the most important factor responsible for the impairment of renal function. The use of the mycophenolate mofetil, a new immunosuppressive agent, allowing a reduction in the dose of cyclosporine, could minimize renal dysfunction. While awaiting the results of a prospective long-term study, close drug monitoring is advised. PMID- 15787795 TI - The sensitized pediatric heart transplant candidate: causes, consequences, and treatment options. AB - Sensitization to HLA antigens and the subsequent development of HLA antibodies in children under consideration for heart transplantation is a significant impediment to survival after listing. This is related both to the frequent need for prospective donor-specific crossmatching (thus limiting donor availability and increasing pretransplant morbidity and mortality), and to the increased risk of adverse outcomes after transplantation. This article will review the scope of this problem in children under consideration for heart transplantation, the different methods available for diagnosing HLA sensitization, the known causes of HLA sensitization, the consequences of these preformed antibodies on outcomes before and after heart transplantation, and the different methods of preventing and treating this sensitization that are currently available. Improved methods of diagnosing, preventing, and treating this problem can only lead to better outcomes for children who require heart transplantation. PMID- 15787796 TI - Hypogammaglobulinemia in pediatric liver transplant recipients. AB - Hypogammaglobulinemia has been reported after solid organ transplantation in adults, however immunoglobulin replacement [intravenous immunoglobulins (IVIG)] is only necessary in a minority of affected patients. We here present three pediatric patients with severe post-transplant hypogammaglobulinemia following liver transplantation (LTx) receiving a cyclosporine-based standard immunosuppression. Patient 1 was transplanted at the age of 10 months for biliary atresia. Eight weeks post-Ltx the serum IgG was 1.7 g/L. Patient 2 was transplanted at the age of 12 yr for acute liver failure. Four weeks post-Ltx the IgG dropped to 2.6 g/L. Patient 3 was transplanted at the age of 4 months for biliary atresia. Ten weeks post-Ltx severe hypogammaglobulinemia (IgG < 1.48 g/L) was diagnosed during a severe infectious complication. Patients 1 and 3 received a steroid bolus therapy for acute graft rejection. All patients had normal IgG concentrations prior to Ltx and lymphocyte subsets were post-operatively in the normal range. There was no extensive loss of protein by ascites. IGIV were replaced in the three patients monthly without further complications. In two of the patients (1 and 3) IVIG therapy was discontinued 8 and 10 months after Ltx when the immunosuppression has been reduced and serum IgG concentrations were found in the normal range without further immunoglobulin replacement. Severe hypogammaglobulinemia is a rare phenomenon following pediatric LTx and seems to be mainly caused by immunosuppressive drugs, however, the exact underlying mechanisms are unclear. A screening for hypogammaglobulinemia is useful after pediatric LTx, especially in patients with an intensified immunosuppression. Moreover, further immunologic research in affected patients is necessary. PMID- 15787797 TI - Risk factors for fungal infection in paediatric liver transplant recipients. AB - Fungal infection (FI) is a major and potentially fatal complication in liver transplantation (LT). Published experience of FI in paediatric LT is limited. We therefore reviewed case records of 79 children, aged between 0.16 and 16 yr, who underwent LT between 1997 and 1998 to document the incidence of, and identify risk factors for, FI. Sixty-eight pre-, peri- and post-LT variables were assessed in relation to FI by univariate and multivariate analyses. The major indications for LT were biliary atresia in 26 (33%) patients, fulminant hepatic failure in 16 (20%) and intrahepatic cholestasis in 11 (14%); eight patients required re-LT. Thirty-two (40.5%) children developed a FI within 1 yr of LT. The median time to FI was 42 days (range 1-342 days). Candida spp. caused 29 (90.7%) FIs; 21 (66%) of these were Candida albicans. Although FI was associated with increased mortality, most patients responded well to antifungal treatment. The variables independently associated with FI were pre-LT fungal colonization and pyrexia and, post-LT, bacterial infection, Epstein-Barr virus (EBV) infection and tacrolimus administration. Identifying risk factors for FI should contribute to the development of strategies for prophylaxis or preemptive therapy. PMID- 15787798 TI - Primary focal segmental glomerulosclerosis--long-term outcome after pediatric renal transplantation. AB - Recurrence of the primary disease is a significant issue in pediatric renal transplantation (RTx). According to data reported by the North American Pediatric Renal Transplantation Cooperative Study, patients with focal segmental glomerulosclerosis (FSGS) as primary renal disease have a recurrence rate of 30% after the first RTx. The relative risk of an early graft loss because of recurrent disease is increased to 1.6-3.1 in pediatric patients with FSGS. In a German open multicenter study, which was initiated to investigate mycophenolate mofetil (MMF) after pediatric RTx [Transplantation 2001:71:638, Transplantation 2003:75:454], patients with FSGS were evaluated for recurrence rate, risk factors for recurrence, long-term graft function, glomerular filtration rate and transplant survival. All patients received immunosuppression with MMF, cyclosporine A and prednisone without induction therapy. Renal function and survival data for FSGS patients were compared with the results of patients with other primary renal diseases within the same study population. Among 86 patients transplanted between 1996 and 1999 eight patients suffered from FSGS as primary disease. Recurrence was diagnosed in two of the eight patients. One out of these two patients lost his graft as a result of recurrence. Risk factors such as time between diagnosis and end stage renal disease (ESRD) and age at onset did not predict recurrence. A three-year patient survival in the FSGS group was 100%, graft survival 87% vs. 97% in the non-FSGS group. Acute rejections occurred in three out of eight FSGS patients and in 37 out of 78 among the non-FSGS group. Long-term renal function, calculated using mathematical modeling based on glomerular filtration rate (GFR) data during 3 yr after RTx, was similar in FSGS patients - including a patient who had recurrence with a functioning graft - and those without FSGS. In patients with FSGS, recurring disease after RTx remains an important cause of graft loss (one of two patients in this population) even under modern immunosuppressants. Nevertheless, the immunosuppressive regimen used was associated with a similar graft survival rate and long-term renal function of FSGS patients compared with patients with other primary diseases. PMID- 15787800 TI - Bilateral lung transplant in Gauchers type-1 disease. AB - We report a case of a patient who received a bilateral lung transplant for end stage lung disease secondary to Gauchers type-1 disease with no evidence of recurrence of the disease in the transplanted lung. PMID- 15787799 TI - A prospective trial of steroid withdrawal after renal transplantation treated with cyclosporine and mizoribine in children: results obtained between 1990 and 2003. AB - A prospective trial of adrenocorticostertoid (steroid) withdrawal after pediatric renal transplantation was begun in 1990. Ninety-four pediatric renal transplant recipients were enrolled in our multicenter study. Immunosuppressive therapy with cyclosporine (CyA), methylprednisolone (MPL), and mizoribine (MZ) was started after transplantation. MPL was reduced to administration on alternate days in 69 patients (73.4%) and was withdrawn in 27 patients (28.7%). Rejection episodes occurred in nine patients (33.3%) after withdrawal of MPL. It occurred within 3 months after withdrawal of MPL in two patients and more than 6 months in the others. Among them, two patients lost the grafts. Thirteen-year patient survival rate and graft survival rate were 94.6 and 83.1%, respectively. Forty-four of the 94 patients reached their final height. Mean final height was 155.0 cm in males and 146.3 cm in females and their height standard deviation score was -2.6 s.d., the same as that at the time of transplantation. Management of growth retardation before transplantation and further reduction in the steroid dose after transplantation will increase the final height of children with chronic renal failure. PMID- 15787801 TI - Carcinoma of donor origin after liver-intestine transplantation in a child. AB - Tumor-related complications after intestinal transplantation in children have been principally EBV driven post-transplant disorders. We describe the clinical course of a child, with a diagnosis of microvillus inclusion disease who received a liver and intestine allograft at the age of 9 months. His postoperative course was significant for multiple episodes of acute intestinal allograft rejection and eventually the development of post-transplant lymphoproliferative disorder (PTLD), which resolved. At 8 yr post-transplant he presented with masses in the intestine allograft mesentery and in the right lobe of the allograft liver, biopsy of which revealed a relatively undifferentiated tumor, suggestive of a carcinoma. In situ hybridization for X and Y chromosomes, revealed his tumor to be of donor origin. Treatment included debulking of the mesenteric mass with segmental enterectomy of the intestinal allograft, and stopping his immunosuppression for a period of 4 months; this resulted in complete resolution of his malignancy. Immunosuppression with tacrolimus and steroids was restarted because of intestinal allograft rejection; he died suddenly of unknown causes at 17 months post-diagnosis of carcinoma. The severely immunosuppressed state produced in this patient allowed for the development of an unusual donor derived carcinoma, which resolved spontaneously with withdrawal of immunosuppression. The mechanism of such regression of tumor may be related to restitution of immunologic competence, but is yet to be determined. PMID- 15787802 TI - Rapid response to rituximab in a pediatric liver transplant recipient with post transplant lymphoproliferative disease and maintenance with sirolimus monotherapy. AB - A 12-yr-old girl with end-stage renal disease secondary to primary hyperoxaluria type I received a living related (left lateral segment) liver transplant from her brother as the first step of a staged liver and kidney transplant. Renal transplantation was planned for a later date from the same donor. Nine weeks after transplantation she developed polymorphic PTLD of the tonsils and adenoids. Initial treatment with surgical resection and withdrawal of immunosuppression was insufficient as she developed recurrence of the PTLD lesion 1 wk after surgical resection and reduction of immunsuppression. Treatment with the chimeric monoclonal anti CD20 antibody, rituximab (Mabthera, Hoffman-La Roche AG, Grenzach Whylen, Germany), resulted in quick response and complete recovery from PTLD within 2 wk, with no recurrence up to 8 months after treatment. Rejection prophylaxis was successfully achieved with Sirolimus (Rapamune, Wyeth Pharmaceuticals Inc., Philadelphia, PA, USA) monotherapy, with no episodes of acute rejection. PMID- 15787803 TI - Caspofungin for invasive fungal infections: combination treatment with liposomal amphotericin B in children undergoing hemopoietic stem cell transplantation. AB - Invasive fungal infections often prove difficult to eradicate especially in the stem cell transplant setting. Amphotericin has been the mainstay of treatment for years but has significant toxicity. Newer antifungal agents, such as caspofungin, have shown promising results in adults, particularly when used in combination with amphotericin as both drugs differ in their mode of action. However, there are few data from children and no previous published information about the use of Caspofungin after paediatric stem cell transplantation. We report our experience in children with proven invasive fungal infections after stem cell transplantation. This combination was non-toxic, and two of three patients survived their infections. PMID- 15787804 TI - An unusual case of pyelonephritis in a pediatric renal transplant recipient. AB - Emphysematous (Gas-forming) pyelonephritis (EPN) is a rare and life-threatening infection of the renal parenchyma with or without involvement of the peri-renal tissues. Diagnosis of this condition is usually made on clinical and radiological grounds. Risk factors for this condition include diabetes mellitus, urinary tract obstruction, renal anomalies such as polycystic kidneys, stones, old age and others. Review of the literature did not reveal any reports of (EPN) in the pediatric age group especially in transplanted patients. We report the first case of EPN in a child, a 12-yr-old boy with a renal transplant, and discuss the various diagnostic and management issues of this particular case and EPN in general. PMID- 15787805 TI - Amylopectinosis disease isolated to the heart with normal glycogen branching enzyme activity and gene sequence. AB - We report a 17-month-old female patient with a rare cause of cardiomyopathy secondary to accumulation of amylopectin-like material (fibrillar glycogen) isolated to the heart. Evidence of amylopectinosis isolated to cardiac myocytes in this patient was demonstrated by histology and electron microscopy. Glycogen content, glycogen branching enzyme (GBE) activity, as well as phosphofructokinase enzyme activities measured in liver, skeletal muscle, fibroblasts and ex transplanted heart tissue were all in the normal to lower normal ranges. Normal skeletal muscle and liver tissue histology and GBE activity, normal GBE activity in skin fibroblasts, plus normal GBE gene sequence in this patient exclude the classical branching enzyme deficiency (type IV GSD). We believe that this is an as yet uncharacterized and novel phenotype of GSD associated with cardiomyopathy, in which there is an imbalance in the regulation of glycogen metabolism limited to the heart. PMID- 15787807 TI - Noninvasive monitoring for methotrexate hepatotoxicity. PMID- 15787809 TI - Regulation of vascular endothelial growth factor expression by insulin-like growth factor-II in human keratinocytes, differential involvement of mitogen activated protein kinases and feedback inhibition of protein kinase C. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is overexpressed in hyperproliferative diseases such as psoriasis and cancer, which are characterized by an increased angiogenesis. It was reported that insulin-like growth factor (IGF)-II is highly expressed during hepatocarcinogenesis and is increased in psoriatic lesions. The increase in IGF-II is believed to be associated with the pathogenesis of these diseases by increasing angiogenesis. OBJECTIVES: In order to investigate the relationship between IGF-II and angiogenesis-related VEGF, VEGF expression in the IGF-II-treated human keratinocytes was monitored and the IGF-II signalling pathways were examined with respect to VEGF expression. METHODS: Northern blot analysis for the VEGF mRNA levels and an enzyme-linked immunosorbent assay for the VEGF protein were performed to determine if IGF-II (100 ng mL(-1)) can increase the VEGF expression levels with or without a pretreatment with protein inhibitors in primary normal human keratinocytes and HaCaT cells. RESULTS: The mRNA and protein levels of VEGF by IGF-II were increased in a time-dependent manner and reached the maximum level 2 h and 8 h after the IGF-II treatment, respectively. However, this increase was abrogated by pretreatment with an extracellular signal-regulated kinase (ERK) inhibitor but not by a p38 inhibitor. The IGF-II-mediated VEGF induction was also effectively inhibited by a pretreatment with the tyrosine kinase inhibitor and Src inhibitor. The PI3-kinase inhibitor also inhibited the expression of VEGF by IGF-II. However, the phospholipase C (PLC) and protein kinase C (PKC) inhibitors did not block the increases of VEGF mRNA level and its protein level by IGF-II, and the PKC inhibitor instead increased VEGF expression by IGF-II. CONCLUSIONS: These results suggest that the tyrosine kinase-Src-ERK1/2 pathway and the PI3-kinase pathway are involved in IGF-II-mediated VEGF expression, but PKC is negatively associated in the IGF-II-mediated VEGF expression. PMID- 15787808 TI - The immunomodulatory effects of regulatory T cells: implications for immune regulation in the skin. AB - Regulatory T cells are thought to have a critical role in the suppression of immune responses. In addition to the prevention of the development of autoimmunity, they are also thought to have a role in the prevention of allergic responses to environmental allergens, immune responses to tumours and the development of memory responses to chronic infections. They have been isolated within the skin and have been shown to express surface markers that enable skin specific migration, suggesting that regulatory T cells have a functional role in the skin. There is accumulating evidence to suggest that regulatory T cells may be involved in numerous skin disorders and may also be modified by various therapeutic agents used to treat these disorders. We review the evidence for the presence of this T-cell subset in humans, the suppressive effects of regulatory T cells, and their role in the skin. PMID- 15787810 TI - Glucosylceramide accumulates preferentially in lamellar bodies in differentiated keratinocytes. AB - BACKGROUND: Sphingolipids, e.g. ceramide (Cer), glucosylceramide (GlcCer) and sphingomyelin (SM), are important bulk constituents of plasma membranes in mammalian cells. In addition, these lipids are also enriched in certain intracellular organelles, as well as in the epidermal lamellar bodies (LBs) of differentiating keratinocytes (KCs). Epidermal Cer, which comprises a heterogeneous family of at least 10 members, is a key component of the stratum corneum (SC) lipids, and regulates permeability barrier function. Levels of GlcCer, but not SM, significantly increase during epidermal differentiation, and then both GlcCer and SM are enzymatically hydrolysed to Cer at and just above the transition from the stratum granulosum to the SC. OBJECTIVES: To determine: (i) whether the GlcCer contained in different pools, i.e. the membrane fraction or the LB fraction, has different metabolic fates; and (ii) whether specific molecular species of GlcCer localize to distinct subcellular pools. METHODS: To study the metabolic fate of specific molecular fractions of GlcCer and Cer, we first ascertained the full spectrum of molecular species present in cultured normal human KCs (CHK) in a differentiated condition as shown in vivo in epidermis. Cer species were analysed in CHK using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Next, the metabolic fate of the GlcCer was studied by pulse-labelling of CHK with L-[14C]-serine. RESULTS: The GlcCer of undifferentiated KCs comprised GlcCer B and Cer NS (or Cer 2), which contain nonhydroxy fatty acid (FA) as the amide-linked FA, while differentiated KCs displayed further heterogeneity of both GlcCer and Cer, including the presence of acylGlcCer and acylCer. The metabolic fates of these sphingolipids were determined. The GlcCer B level decreased over 2 days and then plateaued between days 3 and 5 following pulse-labelling of sphingolipids for 24 h. As GlcCer B declined, Cer NS (Cer 2) increased in a similar time-dependent manner. In contrast, both acylGlcCer and acylCer increased continuously in parallel over this experimental period. CONCLUSIONS: Distinct GlcCer pools segregated to those that were either hydrolysable or nonhydrolysable in differentiated KCs. We assume that the latter pool appears to be LB enriched, and also sequestrates acylGlcCer from other cellular membrane fractions. PMID- 15787811 TI - Retinoic acid suppresses telomerase activity in HSC-1 human cutaneous squamous cell carcinoma. AB - BACKGROUND: Activation of telomerase is crucial for the continued growth and progression of cancer cells. In a previous study, we showed that telomerase is frequently activated in skin tumours. OBJECTIVE: Because retinoic acid (RA) plays an important role in the growth and differentiation of keratinocytes and as RA has some preventive and therapeutic effects on human skin cancers, we examined the effect of RA on the telomerase activity of HSC-1 human cutaneous squamous cell carcinoma cells. RESULTS: Treatment of HSC-1 cells with all-trans RA (ATRA) significantly suppressed their telomerase activity. The suppression of telomerase activity was obvious at day 4 and was maximal at day 5 after the start of treatment with RA. This suppression was reversible as removal of ATRA allowed the recovery of telomerase activity. The suppression of telomerase activity correlated with the decreased expression of mRNA of human telomerase catalytic subunit (hTERT), the rate-limiting determinant of enzyme activity. The production of c-myc and of Sp1 proteins, transcription factors regulating hTERT expression, was not suppressed in HSC-1 cells by ATRA, but phosphorylation of extracellular signal-regulated kinases (ERK)1/2 and of the serine/threonine kinase Akt was significantly suppressed. Phosphorylation of the epidermal growth factor receptor, which regulates hTERT expression in HSC-1 cells, was not altered by ATRA. CONCLUSIONS: These data indicate that RA is effective in inhibiting telomerase activity in HSC-1 cells. Suppression of ERK1/2 and Akt activation is presumed to be involved in the RA-induced suppression of hTERT. PMID- 15787812 TI - Replacement of routine liver biopsy by procollagen III aminopeptide for monitoring patients with psoriasis receiving long-term methotrexate: a multicentre audit and health economic analysis. AB - BACKGROUND: Patients receiving long-term methotrexate for psoriasis are at risk of developing hepatic fibrosis. Repeated liver biopsy has long been regarded as the only reliable method of detecting this and it is still recommended by the American Academy of Dermatology (AAD). More recently, monitoring by serum procollagen III aminopeptide (PIIINP) measurement (Orion Diagnostica, Espoo, Finland) has been advocated as a means of significantly reducing the need for liver biopsy. OBJECTIVES: To assess the validity of guidelines developed in Manchester for the use of PIIINP to monitor patients with psoriasis receiving long-term methotrexate; to assess the anticipated benefits to patients of introducing this change in practice, including reduction in requirement for liver biopsy; and to determine the impact of its introduction on healthcare costs. METHODS: A multicentre audit was conducted over a 24-month period to compare the healthcare costs and outcomes of two intervention groups from centres where serial PIIINP measurement was employed with those of two control groups from centres in which AAD guidelines were followed. RESULTS: A sevenfold reduction in the need for liver biopsy was observed in the two intervention groups (n = 166; 0.04 and 0.02 biopsies/patient/year, respectively) compared with the two control groups (n = 87; 0.26 and 0.30 biopsies/patient/year, respectively). Abnormalities of sufficient severity to influence management were identified in one in five patients biopsied in the main intervention group compared with one in 16 in the control groups. The overwhelming majority of patients surveyed expressed a preference for being monitored by methods that would minimize the need for liver biopsy. The adoption of PIIINP for monitoring would result in significant cost savings. CONCLUSIONS: This audit has shown that patients managed by the Manchester protocol using serial PIIINP measurement and selective liver biopsy were not disadvantaged in comparison with those managed according to AAD guidelines; they were subjected to sevenfold fewer liver biopsies without evidence that important liver toxicity was missed in the process. If PIIINP monitoring were widely adopted, methotrexate would become a more acceptable option for many patients who are dissuaded from considering it because of the threat of repeated liver biopsy; it would also result in significant savings to the healthcare budget. PMID- 15787814 TI - Health status and plasma dioxin levels in chloracne cases 20 years after the Seveso, Italy accident. AB - BACKGROUND: The Seveso, Italy accident of 1976 exposed a large population to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or simply dioxin). The accident resulted, mostly among children, in one of the largest ever-reported outbreaks of chloracne, the typical skin disorder due to halogenated-hydrocarbon compounds. OBJECTIVES: Approximately 20 years after the accident, we conducted an epidemiological study in Seveso to investigate (a) the health status of chloracne cases; (b) TCDD-chloracne exposure-response relationship; and (c) factors modifying TCDD toxicity. METHODS: From 1993 to 1998, we recruited 101 chloracne cases and 211 controls. Trained interviewers administered a structured questionnaire assessing, among other epidemiological variables, information on an extensive list of diseases. During the interview, individual pigmentary characteristics were determined. We measured plasma TCDD levels using high resolution gas chromatography/mass spectrometry. RESULTS: Plasma TCDD was still elevated (> 10 ppt) in 78 (26.6%) of the 293 subjects with adequate plasma samples, particularly in females, in subjects who had eaten home-grown animals, and in individuals with older age, higher body mass index and residence near the accident site. After 20 years, health conditions of chloracne cases were similar to those of controls from the Seveso area. Elevated plasma TCDD was associated with chloracne [odds ratio (OR) = 3.7, 95% confidence interval (CI) 1.6-8.8, adjusted for age, sex and residence]. Chloracne risk was higher in subjects younger than 8 years at the accident (OR = 7.4, 95% CI 1.8-30.3) and, contrary to previous hypotheses, did not increase at puberty onset or in teenage years. Subjects with elevated TCDD levels and light hair colour had higher relative odds of chloracne (OR = 9.2, 95% CI 2.6-32.5). CONCLUSIONS: Dioxin toxicity in chloracne cases was confined to the acute dermatotoxic effects. Chloracne occurrence appeared related to younger age and light hair colour. Age-related dioxin elimination or dilution must be taken into account in interpreting these results. PMID- 15787813 TI - Monitoring patients on methotrexate: hepatic fibrosis not seen in patients with normal serum assays of aminoterminal peptide of type III procollagen. AB - BACKGROUND: The aminoterminal peptide of type III procollagen (PIIINP) is formed during the synthesis of type III collagen and can be measured in the serum. It has been used as a marker for hepatic fibrosis in patients on long-term methotrexate and it has been suggested that serial assay of PIIINP could reduce or eliminate the need for liver biopsies in these patients. OBJECTIVES: To determine whether routine use of the PIIINP assay in a cohort of patients on methotrexate would reliably identify those who were developing hepatic fibrosis and exclude those who were not, thereby reducing or eliminating the need for liver biopsies in this latter group. METHODS: Data were available from a clinical series of 38 patients on methotrexate, who had undergone a total of 70 liver biopsies and 306 PIIINP assays. Liver biopsies were graded using the Roenigk classification. RESULTS: In 34 patients, the findings on 46 liver biopsies could be compared with the results of contemporaneous PIIINP assays. Apart from two biopsies from two patients where fibrosis was no longer detected on a subsequent biopsy, all four biopsies showing fibrosis had abnormal results on over half of the associated PIIINP assays. There were no biopsies showing fibrosis where all associated PIIINP assays were normal. However, 50% of biopsies without fibrosis had at least one abnormal associated assay. In 23 patients, the results of serial PIIINP assays performed between two sequential liver biopsies were correlated with changes in the biopsy in terms of fibrosis. Data were available for 32 pairs of liver biopsies. Apart from a biopsy pair in one patient where fibrosis on the second biopsy was not detected on a third biopsy, all four biopsy pairs defined as showing deterioration had abnormal results on over half of the intervening PIIINP assays. There were no biopsy pairs showing deterioration where all intervening assay results were normal. However, 63% of stable biopsy pairs had at least one abnormal intervening assay. Two patients with nonalcoholic steatohepatitis, which manifests a pattern of fibrosis not scored under the Roenigk classification, had persistently and substantially elevated PIIINP levels. CONCLUSIONS: The data presented support the view that follow-up liver biopsies, as recommended by published guidelines, for patients on long-term low dose methotrexate can be avoided if PIIINP levels are consistently normal. This approach would have reduced the number of patients requiring biopsy in our series by 45%. The PIIINP assay will also be helpful in the management of patients on methotrexate in whom liver biopsy is contraindicated, and in patients with nonalcoholic steatohepatitis. PMID- 15787815 TI - Treatment of female pattern hair loss with oral antiandrogens. AB - BACKGROUND: It has not been conclusively established that female pattern hair loss (FPHL) is either due to androgens or responsive to oral antiandrogen therapy. OBJECTIVES: To evaluate the efficacy of oral antiandrogen therapy in the management of women with FPHL using standardized photographic techniques (Canfield Scientific), and to identify clinical and histological parameters predictive of clinical response. METHODS: For this single-centre, before-after, open intervention study, 80 women aged between 12 and 79 years, with FPHL and biopsy-confirmed hair follicle miniaturization [terminal/vellus (T/V) hair ratio < or = 4 : 1] were photographed at baseline and again after receiving a minimum of 12 months of oral antiandrogen therapy. Forty women received spironolactone 200 mg daily and 40 women received cyproterone acetate, either 50 mg daily or 100 mg for 10 days per month if premenopausal. Women using topical minoxidil were excluded. Standardized photographs of the midfrontal and vertex scalp were taken with the head positioned in a stereotactic device. Images were evaluated by a panel of three clinicians experienced in the assessment of FPHL, blinded to patient details and treatment and using a three-point scale. RESULTS: As there was no significant difference in the results or the trend between spironolactone and cyproterone acetate the results were combined. Thirty-five (44%) women had hair regrowth, 35 (44%) had no clear change in hair density before and after treatment, and 10 (12%) experienced continuing hair loss during the treatment period. Ordinal logistic regression analysis to identify predictors of response revealed no influence of patient age, menopause status, serum ferritin, serum hormone levels, clinical stage (Ludwig) or histological parameters such as T/V ratio or fibrosis. The only significant predictor was midscalp clinical grade, with higher-scale values associated with a greater response (P = 0.013). CONCLUSION: Eighty-eight percent of women receiving oral antiandrogens could expect to see no progression of their FPHL or improvement. High midscalp clinical grade was the only predictor of response identified. A placebo-controlled study is required to compare this outcome to the natural history of FPHL. PMID- 15787816 TI - Corticotropin-releasing hormone triggers differentiation in HaCaT keratinocytes. AB - BACKGROUND: Corticotropin-releasing hormone (CRH) is proposed to be involved in the regulation of the proliferative capacity of keratinocytes, based on its significant actions in the skin. These are mediated by CRH-R1alpha and represented by adenylate cyclase activation, Ca2+ influx, inhibition of cell proliferation and modifications in intracellular signal transduction by NF kappaB. OBJECTIVES: To define CRH action in the cell cycle we investigated its effects on the differentiation programme using the HaCaT keratinocytes model. METHODS: HaCaT keratinocytes were incubated with CRH in Dulbecco's modified Eagles's medium (containing 1.8 mmol L(-1) calcium) or EpiLife (containing 0.06 mmol L(-1) calcium) medium. Cell proliferation was assessed with the MTT assay. Flow cytometry was used for the measurement of DNA content, cell size and granularity and the expression of cytokeratin 14, cytokeratin 1 and involucrin. The electrophoretic mobility shift assay was used to determine DNA binding activity by AP-1 transcription factor. Expression of cytokeratin 1 was also assessed with immunofluorescence microscopy. RESULTS: CRH did produce inhibition of proliferation, which was dose-dependent; the shape of the inhibition curve was determined by the media calcium concentration. CRH action was pinpointed at inhibition of the G0/1 to the S phase transition of the cell cycle. CRH also increased AP-1 binding activity, cell granularity, cytokeratin 1 and involucrin expression, and inhibited cytokeratin 14 expression. CONCLUSIONS: These results are consistent with CRH induction of the keratinocyte differentiation programme. Thus, the overall CRH cutaneous actions connote protective functions for the epidermis, that appear to include the triggering or acceleration of the differentiation programme. PMID- 15787817 TI - Predictions of skin cancer incidence in the Netherlands up to 2015. AB - BACKGROUND: Skin cancer is an important, growing public health problem among white caucasians, causing a heavy burden on dermatologists and general practitioners. OBJECTIVES: To predict the future incidence of skin cancer in the Netherlands up to 2015. METHODS: Expected numbers of skin cancer cases in the Netherlands up to 2015 were calculated by trend modelling of observed rates for melanoma and squamous cell carcinoma (SCC) between 1989 and 2000 obtained from the Netherlands Cancer Registry and for basal cell carcinoma (BCC) obtained from the Eindhoven Cancer Registry; these rates were then multiplied by the predicted age distributions. Incidence rates were fitted to four different models, and predictions were based on the best fitting model. RESULTS: An increase of 80% in the total number of skin cancer patients is expected in the Netherlands: from 20 654 in 2000 to 37 342 in 2015. The total number of melanoma cases is expected to increase by 99%, with the largest increase for males (males aged 35-64, 111%; males aged > or = 65, 139%). Numbers of patients with SCC will increase overall by 80%, mainly among older males and females (increase of 79%) and females aged 35-64 (increase of 93%). The number of cases of BCC will increase by 78%, with the largest increase for the combined groups, those aged 15-64 (males, 66% increase; females, 94% increase), especially for sites other than the head and neck. The contribution of demographic changes (ageing effect) was largest for males with BCC and SCC (35-44%). CONCLUSIONS: If incidence rates for skin cancers in the Netherlands continue to increase and population growth and ageing remain unabated, a rise in annual demand for care of more than 5% could occur, putting a heavy burden on general practitioners and dermatologists. In the absence of marked changes in current ultraviolet radiation exposure, these increases will probably continue after 2015. PMID- 15787818 TI - Gene expression profiles in psoriasis: analysis of impact of body site location and clinical severity. AB - BACKGROUND: Psoriasis is characterized by symmetry of plaques and modulation of multiple genes within those plaques. OBJECTIVES: We compared gene expression profiles of plaques of psoriasis at different anatomical sites for both symmetrical and asymmetrical disease to ascertain whether the same genes were expressed. METHODS: Gene expression profiles were analysed in biopsies from lesional and uninvolved skin from two groups of patients with either predominantly symmetrical or truncal plaques of psoriasis vulgaris, and from normal skin of healthy volunteers. Genomic analyses were performed using cDNA array and kinetically monitored reverse transcriptase-initiated polymerase chain reaction (kRT-PCR) approaches. A cluster of genes upregulated in involved psoriasis skin as compared with normal skin was identified using each of these two technologies. RESULTS: Clustering of patients based on their gene expression profile did not reveal any correlation with family history of psoriasis, age at onset or association of psoriasis with arthritis. There was no difference in gene expression profile between the type (symmetrical vs. truncal) or location (left vs. right side of body) of psoriatic plaques. Gene expression profiles of involved psoriatic skin analysed by kRT-PCR analysis did correlate with both global (Psoriasis Area and Severity Index) and local (erythema, desquamation and plaque elevation) clinical severity. CONCLUSIONS: These results indicate that it may be feasible to analyse the molecular effects of pharmacological agents on psoriatic skin in 'minizone' protocols, that the obtained data can be correlated with clinical severity and that plaques of psoriasis in the same individual express the same genes. PMID- 15787819 TI - In vivo quantification of the structural abnormalities in psoriatic microvessels before and after pulsed dye laser treatment. AB - BACKGROUND: Microvascular abnormalities (capillary elongation, widening and tortuosity) are a characteristic feature of psoriasis and form one of the pathological diagnostic criteria. These changes occur early in the progression of a psoriatic plaque, before there is clinical or histological evidence of epidermal hyperplasia. Treatment of psoriatic microvessels with a pulsed dye laser (PDL) has been associated with both clinical improvement and clearance of lesions. OBJECTIVES: To quantify the structural vascular abnormalities in plaque skin using noninvasive techniques in vivo. Investigations were carried out before and after PDL treatment to determine the nature of laser-induced microvascular changes and the relationship between these changes and clinical improvement. METHODS: Plaque microvessels were visualized using native capillaroscopy. Plaques were then treated three times with the PDL at 14-day intervals. Native capillaroscopy was repeated at 2 and 6 weeks after the final laser treatment. Images were analysed using a combination of nonstereological and stereological measurements. RESULTS: Whole body disease was stable. Treated plaques showed a 48% reduction in plaque severity score (P < 0.01). Native studies showed that the PDL significantly reduced plaque microvessel density (P < 0.05), image area fraction (P < 0.01), microvessel length density (P < 0.01) and vessel image width (P < 0.01). The reduction in plaque severity score (which denoted clinical improvement) was related quantitatively to the reduction in microvessel area per unit area of plaque skin, i.e. the image area fraction (correlation coefficient = 0.772, P < 0.01). The greatest response of plaque microvessels was within 2 weeks after the final laser treatment, while the greatest reduction in plaque severity score occurred between 2 and 6 weeks after the final laser treatment, i.e. clinical improvement was preceded by microvascular improvement. CONCLUSIONS: These findings indicate that there is a close correlation between the state of the superficial vasculature and the clinical status of psoriasis. The expanded superficial microvascular bed in plaque skin is a necessary component for maintaining clinical lesions and these blood vessels are thus a legitimate target for treatment. PMID- 15787820 TI - Association of melanoma and neurocutaneous melanocytosis with large congenital melanocytic naevi--results from the NYU-LCMN registry. AB - BACKGROUND: Large congenital melanocytic naevi (LCMN), which develop in utero and are present in approximately one in 20,000 newborns, are associated with markedly increased risks of cutaneous melanoma, leptomeningeal melanoma and neurocutaneous melanocytosis (NCM). OBJECTIVES: This study examined clinical characteristics associated with melanoma and NCM among patients with LCMN, and estimated the risk of developing melanoma and NCM in these patients. METHODS: Two hundred and five LCMN patients enrolled in the New York University registry were studied. One hundred and seventy of these patients were followed prospectively. The remaining 35 patients had either melanoma at the time of entry into the registry (n = 6), or had insufficient follow-up information (n = 29). The outcome measures were the occurrence of melanoma and NCM. The associations between these outcomes and the clinical covariates (anatomical location of the LCMN, size of the LCMN, number of satellite lesions, family history of melanoma, patient sex and treatment) were assessed. RESULTS: Four of 170 (2.3%) prospectively followed patients developed melanomas, representing a standardized morbidity ratio of 324. Among the entire cohort (n = 205), there were associations between increasing numbers of satellite naevi and the occurrence of melanoma (P = 0.04), and the presence of NCM (P = 0.06). Compared with patients who did not develop these diseases, median LCMN diameters were larger among patients who developed melanoma (49 vs. 39 cm) and NCM (55 vs. 46 cm). CONCLUSIONS: In LCMN patients, increasing numbers of satellite lesions and larger LCMN diameters are associated with melanoma and NCM. PMID- 15787821 TI - Oral retinoids for the prevention of skin cancers in solid organ transplant recipients: a systematic review of randomized controlled trials. AB - BACKGROUND: The increased incidence of skin cancers after solid organ transplantation is well recognized. Skin cancers developing in transplant recipients are more aggressive in behaviour. Therapeutic options to reduce and/or delay the development of cutaneous neoplasms are therefore of interest. OBJECTIVES: The objective of this review was to summarize the available medical literature from randomized controlled trials on the use of oral retinoids as a preventive agent for skin cancers in the solid organ transplant population. METHODS: Three electronic databases were searched for relevant trials: MEDLINE (1966-October 2003), EMBASE (1980-week 44, 2003) and the Cochrane Controlled Trials Register (third quarter 2003). Randomized or quasi-randomized controlled clinical trials on subjects of any age or ethnic background who had received a solid organ transplant (cardiac, renal, liver, etc.) were evaluated. All titles and abstracts found by the search strategy were independently reviewed by two researchers for inclusion into the review. RESULTS: Eighty-one abstracts were identified through the electronic databases for consideration. Review of the abstracts identified three eligible trials. One cross-over trial involving 23 subjects treated with acitretin 25 mg daily for 12 months reported 46 squamous cell carcinomas (SCCs) developing in six subjects during acitretin treatment vs. 65 SCCs developing in 15 subjects during the drug-free period. Another trial involving 44 subjects treated with acitretin 30 mg daily or placebo for 6 months reported two of 19 subjects developing two SCCs in the treatment group vs. nine of 19 subjects developing 18 new skin cancers (15 SCCs, one Bowen's disease, two basal cell carcinomas) in the placebo group. One dose comparison trial involving 26 renal transplant recipients treated with acitretin did not find a significant difference in numbers of skin cancers developing at the doses examined. The major limitation to the use of acitretin was poor tolerance due to adverse events. Headaches, rash, musculoskeletal symptoms and hyperlipidaemia were the most common causes of withdrawal from treatment. No alterations in renal or liver function were detected during the periods of treatment or follow-up. CONCLUSIONS: The available data from a small number of randomized controlled trials suggest that acitretin may have a role in the management of solid organ transplant recipients with skin cancers. Tolerability of the drug is a major factor limiting its use. Appropriate selection of patients may help improve the risk-benefit ratio. PMID- 15787822 TI - Pilot study of topical delivery of methotrexate by electroporation. AB - BACKGROUND: The topical administration of methotrexate (MTX) for the treatment of psoriasis and neoplastic diseases is restricted by the poor diffusion of MTX across the stratum corneum. OBJECTIVES: We applied electroporation to increase the transdermal transport of MTX. METHODS: Electrodes were placed either side-by side on the surface of excised full thickness pig skin, or on a piece of skin clamped between compartments of a vertical diffusion chamber. Sixty rectangular electric pulses at 120 V, 1 ms and 1 Hz were applied across the skin. MTX was left on the skin surface for an additional 10 min to take advantage of diffusion through electropores. Cumulative drug transport was measured by radioactive tracing, using [3H]-methotrexate, from punch biopsy samples taken from under the cathode. The integrity of the radioisotope was verified by high-performance liquid chromatography. RESULTS: Using side-by-side electrodes, treatment with the pulses alone resulted in a 2.5-fold increase; adding anionic lipid enhancers to the pulses resulted in a 4.4-fold enhancement compared with passive diffusion. Concurrent iontophoresis for the 11-min time period made a nonsignificant contribution. To reduce tissue resistance we used 40 degrees C hyperthermia in a vertical diffusion chamber; transport was increased 11-fold to 53 microg cm(-2) (flux = 290 microg cm(-2) h(-1)). MTX penetration profiles indicated that more than half of the MTX was confined to the epidermis and papillary dermis. The tissue concentration in this superficial reactive unit was 1.7 mmol L(-1). CONCLUSIONS: Electroporation of MTX with an anion lipid enhancer under a mild hyperthermic environment provided a significant transdermal delivery within a short application time. The method may be an effective means of drug delivery for treating psoriasis or other MTX-sensitive disorders and avoids the potential systemic toxicity. PMID- 15787823 TI - Successful treatment of three cases of nephrogenic fibrosing dermopathy with extracorporeal photopheresis. AB - BACKGROUND: Nephrogenic fibrosing dermopathy (NFD) is a recently described cutaneous fibrosing disorder associated with renal dysfunction. Patients present with thickened skin or oedematous skin with indurated papules and plaques involving extremities and trunk, and often associated with disabling contracture of the adjacent joints. The aetiology and pathogenesis remain largely unknown. As a consequence, therapeutic measures with proven efficacy are nonexistent to date. OBJECTIVES: To consider treatment with extracorporal photopheresis (ECP) in three patients. Patients We report three new cases of NFD with the characteristic clinical and pathological features. Two patients required haemodialysis due to end-stage renal failure, despite prior renal transplantation. One patient had renal dysfunction but was never on dialysis, nor had she been transplanted. ECP treatment was administered at intervals of 2-4 weeks. RESULTS: All three patients showed a softening of the skin lesions and a marked improvement of the joint motility starting after four cycles of ECP. One patient developed a complete regression of her skin lesions after 16 cycles of ECP, and response to therapy was observed despite constantly elevated renal values. CONCLUSION: These data indicate that ECP may represent a valuable therapeutic option for NFD. PMID- 15787824 TI - Bullous pemphigoid antigen 1 isoforms: potential new target autoantigens in multiple sclerosis? AB - BACKGROUND: The simultaneous occurrence of bullous pemphigoid (BP) and multiple sclerosis (MS), two autoimmune diseases involving the skin and the central nervous system (CNS), respectively, has been described. OBJECTIVES: As the BPAG1 gene encodes the epithelial isoform of BP antigen 1 (BPAG1-e), a major autoantigen of BP, as well as additional variants expressed in the neurones of the CNS and peripheral nervous system and in Schwann cells, we tested the hypothesis that products of the BPAG1 gene act as shared autoantigens in both BP and MS. METHODS: The reactivity of cerebrospinal fluid (CSF) obtained from 18 patients with MS, 10 patients with other inflammatory CNS diseases and 20 normal controls was assayed by immunoblotting against two recombinant fragments of BPAG1 e encompassing regions that are also found in the neuronal variants BPAG1-n and BPAG1-a. RESULTS: The recombinant protein glutathione-S-transferase (GST)-BPAG1 e1-887 was recognized by five of 18 (27%) CSF samples obtained from patients with MS, two of 10 (20%) samples from patients with other inflammatory neurological diseases and five of 20 (25%) samples from normal controls. Furthermore, two of 18 (11%) CSF samples from patients with MS bound to GST-BPAG1-e1880-2649, whereas none of the samples obtained from patients with other inflammatory neurological diseases or from control subjects showed reactivity. CONCLUSIONS: These results raise the possibility that a subset of patients with MS develops an autoantibody response to the neuronal variants of BPAG1. These findings potentially open the avenue of neuronal BPAG1 variants being novel targets of autoantibodies in neurological diseases. PMID- 15787825 TI - Intralesional therapy with anti-CD20 monoclonal antibody rituximab: local and systemic efficacy in primary cutaneous B-cell lymphoma. AB - Rituximab (MabThera, Roche) is a chimeric monoclonal antibody directed against the CD20 antigen. Its efficacy and safety were first demonstrated in the treatment of systemic B-cell lymphomas. We report the use of intralesional injections of rituximab into some but not all cutaneous lesions in a patient with multiple primary cutaneous follicular centre B-cell lymphoma. This treatment resulted in tumour regression, even of the lesions that had not been injected. We therefore hypothesize that there is systemic diffusion of rituximab from injected sites despite the low doses injected locally, or the induction of a specific antitumour immune response acting systemically. PMID- 15787826 TI - Incidence of xeroderma pigmentosum in Larkana, Pakistan: a 7-year study. AB - Xeroderma pigmentosum (XP) is a rare autosomal recessive inherited disorder caused by a defect in the normal repair of DNA of various cutaneous cell types damaged by exposure to ultraviolet radiation. We present our 7-year experience with 36 XP patients who either visited the Department of Dermatology or were seen in the medical camps arranged in remote areas for patients' welfare, from 1995 to 2001. For ease of discussion we classified all cases into the following subgroups on clinical grounds only: mild, those with light brown freckles on the face alone; moderate, those with dark brown freckles with burning on the face, neck, ears, chest, hands and photophobia but without other associated obvious cutaneous and ocular changes; severe, those with extensive dark brown freckles with burning on the exposed parts as well as on the unexposed parts of the body, i.e. the chest, back, abdomen and arms including other associated cutaneous and ocular changes such as ulcers and malignancy. Of 36 patients, three (8.3%) were classified as mild, nine (25%) moderate and 24 (66.7%) severe; there were 18 males and 18 females, age range 2-30 years (mean 8.9 years). Seventeen patients had cutaneous changes: actinic keratosis, keratoacanthoma, fissures and ulcerative nodules on the exposed parts of the body. Four patients had wide ulcers, along with mass formation and severe pigmentation on the face, neck and head. Twenty-nine patients developed ocular symptoms: photophobia, conjunctivitis, corneal keratitis and lid ulcer. One patient had complete loss of vision. Histopathological findings revealed that six patients had squamous cell carcinoma (SCC) on the face, head, ear or lip. More than one sibling (two to four) was affected in four families. The majority of cases (20/36, 55.6%) were from the Brohi tribe (skin type III), while the remaining cases (16/36, 44.4%) were from the Sindhi population (skin type IV). The large number of XP patients seen in those with skin type III (Brohi tribe) compared with skin type IV (Sindhi population) indicates that the skin type and the race has a considerable value in the pathogenesis of XP. Furthermore, 24 of 36 patients were in the severe group and six of these had SCC. Moreover, no neurological abnormalities were observed in our patients. All patients were treated according to disease severity by prescribing oral antibiotics, local steroids, sunscreens and/or chemotherapy followed by irradiation in malignant cases. Two patients died because of extensive SCC. PMID- 15787827 TI - Rapid improvement of recalcitrant disseminated granuloma annulare upon treatment with the tumour necrosis factor-alpha inhibitor, infliximab. AB - Granuloma annulare (GA) is a chronic inflammatory disorder of unknown aetiology, which is characterized clinically by erythematous plaques preferentially localized to the distal extremities, although disseminated variants exist. In light of the chronic relapsing nature of GA and lack of satisfactory treatment options, we initiated treatment with infliximab in a patient with chronic disseminated GA that was recalcitrant to standard treatment. The 59-year-old female patient with insulin-dependent diabetes had experienced GA lesions for more than 4 years despite various systemic and topical treatments. Systemic glucocorticoids were not a therapeutic option because of the preexisting unstable insulin-dependent diabetes. Infliximab was administered intravenously at 5 mg kg( 1) day(-1) at weeks 0, 2 and 6 and thereafter at a monthly interval for an additional 4 months. Most of the GA plaques resolved within 4-6 weeks, leaving postinflammatory brownish macules. Newly arising plaques disappeared within 2 weeks and new GA lesions were not observed during the entire observation period of more than 16 months. Infliximab may be an additional option in the treatment of recalcitrant forms of GA as well as in other chronic granulomatous skin disorders, such as sarcoidosis and necrobiosis lipoidica. PMID- 15787828 TI - Androgenetic alopecia in children: report of 20 cases. AB - Androgenetic alopecia (AGA) is the most common type of hair loss in adults. Although there are differences in the age at onset, the disease starts after puberty when enough testosterone is available to be transformed into dihydrotestosterone. We report 20 prepubertal children with AGA, 12 girls and eight boys, age range 6-10 years, observed over the last 4 years. All had normal physical development. Clinical examination showed hair loss with thinning and widening of the central parting of the scalp, both in boys and girls. In eight cases frontal accentuation and breach of frontal hairline were also present. The clinical diagnosis was confirmed by pull test, trichogram and dermoscopy in all cases, and by scalp biopsy performed in six cases. There was a strong family history of AGA in all patients. The onset of AGA is not expected to be seen in prepubertal patients without abnormal androgen levels. A common feature observed in our series of children with AGA was a strong genetic predisposition to the disease. Although the pathogenesis remains speculative, endocrine evaluation and a strict follow-up are strongly recommended. PMID- 15787829 TI - A case of chromoblastomycosis with an unusual clinical manifestation caused by Phialophora verrucosa on an unexposed area: treatment with a combination of amphotericin B and 5-flucytosine. AB - Chromoblastomycosis is a cutaneous and subcutaneous mycotic disease caused by the dematiaceous (black) fungi. Five species of fungi are known generally to be the cause: Fonsecaea pedrosoi, Phialophora verrucosa, Cladosporium carrionii, F. compacta and Rhinocladiella cerphilum. In infected tissue they can appear as pigmented sclerotic bodies, commonly called 'copper pennies', which are pathognomonic of chromoblastomycosis. The infection usually occurs through traumatic skin inoculation, with the majority of lesions occurring on the feet and legs of outdoor workers. We report a patient in whom the lesions had begun on the right breast, which is an unexposed area, without a history of trauma. A uniform, reliable treatment does not exist but our patient was mycologically cured with the use of amphotericin B and the subsequent combination of 5 flucytosine and itraconazole. PMID- 15787830 TI - Lymphomatoid annular erythema: a new form of juvenile mycosis fungoides. PMID- 15787831 TI - Topical liarozole in ichthyosis: a double-blind, left-right comparative study followed by a long-term open maintenance study. PMID- 15787832 TI - Tacrolimus ointment-associated alcohol intolerance in infants receiving ethanol containing medication. PMID- 15787833 TI - Reactivation of ophthalmic herpes zoster following pulsed-dye laser treatment for inflammatory acne vulgaris. PMID- 15787834 TI - Restored epidermal CD44 expression in lichen sclerosus et atrophicus and clinical improvement with topical application of retinaldehyde. PMID- 15787835 TI - A study of patterns of androgenetic alopecia in men: an Indian perspective. PMID- 15787836 TI - Eosinophilic angiocentric fibrosis--a rare mucosal variant of granuloma faciale which may present to the dermatologist. PMID- 15787837 TI - Utility of studying the ocular surface in patients with acne vulgaris treated with oral isotretinoin: a randomized controlled trial. PMID- 15787838 TI - Expression of elafin in extramammary Paget's disease. PMID- 15787839 TI - Teenage acne is influenced by genetic factors. PMID- 15787840 TI - Successful treatment of chronic ulcerated necrobiosis lipoidica with 0.1% topical tacrolimus ointment. PMID- 15787841 TI - Treatment of relapsing idiopathic nodular panniculitis with clofazimine. PMID- 15787842 TI - Urticaria as a side-effect of diphencyprone therapy for resistant viral warts. PMID- 15787843 TI - Fatal fulminant legionnaires' disease in a patient with severe erythodermic psoriasis treated with infliximab after long-term steroid therapy. PMID- 15787844 TI - Tendon destruction induced by Panton-Valentine leukocidin-positive Staphylococcus aureus in a patient with Sezary syndrome. PMID- 15787845 TI - Generalized pustular psoriasis successfully treated with topical tacrolimus. PMID- 15787846 TI - Squamous cell carcinoma arising on scars of epidermolysis bullosa acquisita. PMID- 15787847 TI - Refractory human papillomavirus-associated oral warts treated topically with 1-3% cidofovir solutions in human immunodeficiency virus type 1-infected patients. PMID- 15787848 TI - Hypocalcaemia-induced pustular psoriasis-like skin eruption. PMID- 15787850 TI - Aging in context: age-related changes in context use during language comprehension. AB - Effects of normal aging on the use of sentence context information during language comprehension were examined by measuring younger and older adults' event related potential (ERP) responses to congruent sentence-final words as a function of contextual constraint. Half of the sentence contexts were strongly constraining, rendering the target word very predictable, whereas the other half were weakly constraining. Both age groups elicited smaller N400 responses to target words in strongly than in weakly constrained contexts, but this effect was significantly smaller and later for older adults. Older adults with lower reading spans showed greater delays. Age-related changes were driven primarily by decreases in older adults' ability to make use of the richer information available from strongly constraining contexts to guide semantic processing, as word processing (N400s) in weak contexts was qualitatively and quantitatively similar in the two age groups. PMID- 15787851 TI - The effects of memory scanning on the late Nd and P300: an interference study. AB - Singhal and Fowler (2004) showed that the late negative difference (Nd) waveform elicited during dichotic listening was attenuated by concurrent visual short term memory (STM) scanning, but not long term memory (LTM) scanning. P300 was reduced by both tasks. The present study compared the effects of STM load on the late Nd and P300 by combining dichotic listening and visual memory scanning with varying set sizes. The results showed that the late Nd was sensitive to the introduction of the scanning task, but not to an increase in load. Furthermore, both the auditory and visual P300s were reduced when a second task was introduced, but only the visual P300 decreased as a function of memory-set size. These data suggest that (a) the auditory late Nd reflects working memory, but not memory scanning, (b) late Nd and P300 reflect parallel but distinct working memory processes, and (c) stimulus modality is an important determinant of P300 amplitude. PMID- 15787852 TI - On the ERN and the significance of errors. AB - The error-related negativity (ERN) is an event-related brain potential observed when subjects commit errors. To examine whether the ERN is sensitive to the value of errors, the motivational significance of errors was manipulated in two experiments. In Experiment 1, low and high monetary value errors were compared to evaluate the effect of trial value on the ERN. In Experiment 2, subjects performed a flanker task both while their performance was being evaluated and during a control condition. Consistent with the notion that the error-detection system is sensitive to the significance of errors, the ERN was significantly larger on high-value trials in Experiment 1 and during evaluation in Experiment 2. There were no corresponding effects on the correct response negativity, and no behavioral differences between conditions were evident in either experiment. These results are discussed in terms of the functional role of the ERN in response monitoring. PMID- 15787853 TI - Brain potentials associated with expected and unexpected good and bad outcomes. AB - The error-related negativity (ERN) is an event-related brain potential observed when subjects receive feedback indicating errors or monetary losses. Evidence suggests that the ERN is larger for unexpected negative feedback. The P300 has also been shown to be enhanced for unexpected feedback, but does not appear to be sensitive to feedback valence. The present study evaluated the role of expectations on the ERN and P300 in two experiments that manipulated the probability of negative feedback (25%, 50%, or 75%) on a trial-by-trial basis in experiment 1, and by varying the frequency of positive and negative feedback across blocks of trials in experiment 2. In both experiments, P300 amplitude was larger for unexpected feedback; however, the ERN was equally large for expected and unexpected negative feedback. These results are discussed in terms of the potential role of expectations in processing errors and negative feedback. PMID- 15787854 TI - Covert attention in touch: behavioral and ERP evidence for costs and benefits. AB - To investigate the mechanism underlying tactile spatial attention, reaction times (RTs) and event-related potentials (ERPs) were recorded in response to mechanical stimuli delivered to the hands. At the start of each trial cues indicated either the correct (valid) or incorrect (invalid) tactile stimulus location or were uninformative (neutral). RT costs (suppression of invalid compared to neutral trials) were found to be larger than benefits (enhancement of valid compared to neutral trials). ERPs showed that costs and benefits contribute equally to attentional modulations of the somatosensory N140 component, whereas these were largely due to costs at longer latencies. These results differ from the pattern of attentional ERP modulations previously found for vision and audition, where costs precede benefits, and therefore suggest that the mechanisms of attentional selectivity in touch might be different from attentional processes in other modalities. PMID- 15787855 TI - Fine-tuning of auditory cortex during speech production. AB - The cortex suppresses sensory information when it is the result of a self produced motor act, including the motor act of speaking. The specificity of the auditory cortical suppression to self-produced speech, a prediction derived from the posited operation of a precise forward model system, has not been established. We examined the auditory N100 component of the event-related brain potential elicited during speech production. While subjects uttered a vowel, they heard real-time feedback of their unaltered voice, their pitch-shifted voice, or an alien voice substituted for their own. The subjects' own unaltered voice feedback elicited a dampened auditory N100 response relative to the N100 elicited by altered or alien auditory feedback. This is consistent with the operation of a precise forward model modulating the auditory cortical response to self-generated speech and allowing immediate distinction of self and externally generated auditory stimuli. PMID- 15787856 TI - Expectancy effects on omission evoked potentials in musicians and non-musicians. AB - Abstract An expanded omitted stimulus paradigm was investigated to determine whether expectancy would modulate the amplitude of the omission evoked potentials (OEPs). In addition, we examined the effects of musical expertise on OEPs. Trials started with 3-7 beats randomly and contained 5 omitted beats. Three types of trials (n = 90) were presented with 1, 2, or 3 beats occurring between omissions. A tap response at the end of each trial was used to determine timing accuracy. Clear OEPs were observed over midline sites. We found main omission effects with respect to an N150 and a P400 OEPs component, such that peak amplitudes diminished whenever the occurrence of an omitted stimulus could be expected. In addition, an N600 OEPs component emerged in response to expectedly omitted stimuli toward the end of each trial within the group of musicians. Thus, musical training seems to lead to more efficient and more refined processing of auditory temporal patterns. PMID- 15787857 TI - A genetic analysis of ambulatory cardiorespiratory coupling. AB - This study assessed the heritability of ambulatory heart period, respiratory sinus arrhythmia (RSA), and respiration rate and tested the hypothesis that the well-established correlation between these variables is determined by common genetic factors. In 780 healthy twins and siblings, 24-h ambulatory recordings of ECG and thorax impedance were made. Genetic analyses showed considerable heritability for heart period (37%-48%), RSA (40%-55%), and respiration rate (27% 81%) at all daily periods. Significant genetic correlations were found throughout. Common genes explained large portions of the covariance between heart period and RSA and between respiration rate and RSA. During the afternoon and night, the covariance between respiration rate and RSA was completely determined by common genes. This overlap in genes can be exploited to increase the power of linkage studies to detect genetic variation influencing cardiovascular disease risk. PMID- 15787859 TI - Prepulse inhibition of startle and its moderation by schizotypy and smoking. AB - Abstract The influences of smoking status and schizotypy on prepulse inhibition (PPI) of the startle eye-blink response were assessed in 71 healthy volunteers, across a wide range of prepulse-to-pulse intervals (50-2020 ms). Multiple regression analyses revealed that nonsmoking participants high in cognitive disorganization showed reduced PPI between 50 and 260 ms, whereas at prepulse intervals of 80 and 140 ms individuals high in introvertive anhedonia had greater PPI compared to their low-scoring counterparts. Moreover, there were positive associations in nonsmokers between introvertive anhedonia and latencies to onset and peak response. In contrast, for those individuals who smoked these associations were attenuated or abolished. The results suggest that PPI is altered differentially in psychosis-prone populations who display different symptom profiles, and that these relationships are moderated by smoking status. PMID- 15787858 TI - Effects of baroreceptor stimulation and opioids on the auditory startle reflex. AB - We examined (a) whether carotid baroreceptor stimulation attenuates the auditory startle response and its modulation by preceding affective pictures, and (b) whether these effects are mediated by endogenous opioids. Seventy-eight young normotensive adults with or without a parental history of hypertension received brief exposures to affective pictures and noise bursts during phasic manipulation of the carotid baroreceptors. In each participant, opioids were blocked by naltrexone in half of the sessions. Baroreceptor stimulation had a strong dampening effect on the startle response. This effect was not influenced by opioid blockade, sex, or parental history of hypertension. No baroreceptor effects were obtained regarding ratings of the affective pictures or startle modulation by the pictures. The baroreceptor stimulation effects seem to be mediated by the basal primary acoustic startle circuit rather than by higher affective circuits. PMID- 15787860 TI - Neuroticism, cortisol reactivity, and antibody response to vaccination. AB - This study examined whether neuroticism was related to the antibody response to influenza vaccination and whether the relationship was mediated by cortisol reactions to acute laboratory mental stress. Antibody status was assessed at baseline and to a trivalent influenza vaccination in 57 students at 5-week and 5 month follow-up. Neuroticism was also measured at baseline. Cortisol was measured at rest and in response to a pressurized mental arithmetic task. At both follow ups, higher neuroticism scores were associated with poorer A/Panama antibody response, following adjustment for baseline antibody titer. Higher neuroticism scores were also associated with blunted cortisol reactivity, and blunted cortisol reactivity was associated with poorer A/Panama antibody response, but only at 5 months. However, there was no conclusive evidence that cortisol reactivity mediated the association between neuroticism and antibody response. PMID- 15787861 TI - Psychopathic traits and autonomic responding to concealed information in a prison sample. AB - We examined the role of psychopathic traits on physiological responding during a concealed information polygraph test among prison inmates (n = 40). The Psychopathic Personality Inventory (Lilienfeld & Andrews, 1996) was used to assess psychopathic traits. Cardiac, electrodermal, and respiratory responses were measured while participants were presented with personal and control names and asked to conceal recognition of personal information. We first piloted the present concealed information test in a sample of undergraduates (n = 27). Enhanced orienting to concealed information compared to control information was observed in both prisoners and undergraduates. However, prisoners scoring high on the Impulsive Antisociality factor of the Psychopathic Personality Inventory (PPI II) exhibited reduced electrodermal responding. This finding indicates that the sensitivity of the concealed information test may be decreased in antisocial offenders. PMID- 15787862 TI - Filter properties of root mean square successive difference (RMSSD) for heart rate. AB - The root mean square successive difference (RMSSD) in heart period series is a time domain measure of heart period variability. The RMSSD is sensitive to high frequency heart period fluctuations in the respiratory frequency range and has been used as an index of vagal cardiac control. By transfer function simulations, the RMSSD statistic is shown to represent a high-pass filter that effectively captures respiratory sinus arrhythmia but also passes lower frequency fluctuations that can include sympathetic influences. These simulations, together with analysis of actual heart period series, reveal that the RMSSD is biased by basal heart period. Although between-subjects levels of RMSSD covary highly with spectral estimates of high-frequency variability, within-subject RMSSD change scores account for only 50-60% of the variance in spectral estimates. The present findings raise caveats in the applications and interpretation of the RMSSD statistic. PMID- 15787863 TI - Case reports--what is their value? PMID- 15787865 TI - Population pharmacokinetics of levocetirizine in very young children: the pediatricians' perspective. AB - Developmental changes during infancy and childhood can affect drug pharmacokinetics (PK), i.e., absorption, distribution, metabolism, and renal excretion. This, in turn, influences optimal dosing, efficacy, and safety. To date, of the 40 H1-antihistamines available worldwide, only 11 have been studied in children using a PK approach. Here, we provide the pediatricians' perspective on the population PK of levocetirizine, the pharmacologically active enantiomer of cetirizine, in very young children who received oral cetirizine, and describe the factors that influence levocetirizine PK in this population. In a prospective, randomized, double-blind, parallel-group, placebo-controlled study, very young children received oral cetirizine 0.25 mg/kg twice daily for 18 months. Plasma levocetirizine concentrations were measured in timed, sparse blood samples collected at steady-state (3, 12 and 18 months after commencement of treatment) for the purpose of monitoring safety, and levocetirizine population, PK parameters were derived by using non-linear mixed effects modeling. In 343 children (age 14-46 months, body weight 8.2-20.5 kg), a total of 943 blood samples were obtained. Compliance with cetirizine dosing was documented. The population PK model used predicted that with increasing body weight, levocetirizine oral clearance would increase by 0.044 l/h/kg, and levocetirizine volume of distribution would increase by 0.639 l/kg. Levocetirizine PK were not influenced by eosinophilia, sensitization to allergens, allergic disease, gastroenteritis/diarrhea, or concomitant ingestion of other medications. This population PK model predicts that in very young children, the oral clearance of levocetirizine will be rapid and will increase as body weight and age increase, therefore, levocetirizine dosing should be based on body weight and age in this population. Compared with older patients, on a mg/kg basis, relatively higher doses may be needed, and twice-daily dosing may be necessary, as previously reported for the related racemic H1-antihistamine cetirizine. PMID- 15787866 TI - Putative regulatory T cells are impaired in cord blood from neonates with hereditary allergy risk. AB - The hygiene hypothesis implies that the increasing prevalence of allergy in 'westernized' countries is explained by reduced bacterial exposure in early life, but the underlying mechanism remains elusive. We therefore wanted to study the effect of bacterial lipopolysaccharide (LPS) on the generation of regulatory T (T(R)) cells in neonates, and to analyze differences between neonates with allergy risk because of a family history of atopy (FH+) and controls without such hereditary risk (FH-). Cord blood mononuclear cells from the FH+ and FH- groups were stimulated with beta-lactoglobulin in the presence of LPS. T-cell phenotypes suggestive of T(R) cells [CD25+, CD25high and integrin (CD103+)], and the intracellular proliferation antigen Ki-67 were quantified by flow cytometry. Release of the immunosuppressive cytokine transforming growth factor beta1 (TGF beta1) from its inactive complex was determined by enzyme-linked immunosorbent assay. The analyses revealed the generation of T-cell phenotypes suggestive of T(R) cells including a CD25high T-cell subset which was inversely related to T cell proliferation (r=-0.54, p<0.05) and to activation-induced release of TGF beta1 (r=-0.80, p<0.001). The CD25high T-cell subset tended to be impaired in the FH+ group (% of CD3+ T cells: FH+, 5.1% vs. FH-, 12.6%), and notably, the FH+ group showed a significantly reduced capacity for generation of both CD25+ (FH+, 16.2% vs. FH-, 34.9%; p<0.01) and T cells (FH+, 2.1% vs. FH-, 3.9%; p<0.05). Our findings suggested that early-life exposure to a dietary antigen in the presence of LPS might modulate the immune system by generating T(R) cells. This capacity was impaired in neonates with hereditary allergy risk, but clinical follow-up will be required to determine a possible effect on allergy emergence. PMID- 15787867 TI - Antigen specific cytokine response in pediatric patients with atopic dermatitis. AB - The physiopathology of atopic dermatitis (AD) has still to be elucidated. T effector cells with cutaneous homing receptors or T-cell derived cytokines have been assumed to be implicated in the pathogenetic mechanisms in AD and to be responsible for the different immunologic responses of patients. In fact, the large majority of AD patients display high IgE levels while others do not develop an abnormal IgE response. Although, there are not significant clinical features characterizing the two different groups, patients with normal IgE belong to a younger age range, raising the possibility that the hypothesized dichotomy of AD might be due to age. In the present study we included 172 outpatient children attending the Pediatric Department of our institution. Serum IgE levels and percentages of peripheral T lymphocytes expressing the cutaneous homing antigen (CLA) were evaluated and results were analyzed in relation to the activity of the disease (SCORAD index) or age. In the overall patients, the IgE levels increased significantly with age (0-1 yr: 19.50 IU/ml; 1-3 yr: 62.0 IU/ml; 3-8 yr: 96.0 IU/ml; >8 yr: 148.5 IU/ml; p<0.001) and with the severity of the disease (SCORAD low: 46.80 IU/ml; medium: 42.90 IU/ml; high: 148.5 IU/ml; p=0.01). Percentages of CLA+ peripheral T lymphocytes also increased with age (0-1 yr: 3.3; 1-3 yr: 4.85; 3-8 yr: 10.6; >8 yr: 12.5; p<0.001), although they were not significantly different in patients with different SCORAD (p=0.89). We further investigated the cellular immune response to a specific antigen in 25 subjects, matched for age, SCORAD, and CLA+ T-cell percentages. Among them, 13 patients had casein serum specific IgE and 12 had no evidence of casein sensitization. Peripheral blood mononuclear cells were kept in short-term culture with endotoxin-free casein fractions and IFN-gamma, TNF-alpha, IL-5, IL-10 cytokine-producing cells were detected by ELISpot. Statistical analysis showed significant higher numbers of TNF-alpha- or IL-10-producing cultures (stimulation index >3) in the 'allergic' patients than in the milk tolerant subjects (p=0.01 and 0.05). The analysis of individual responses confirmed this finding but also provide evidence of a significant increase in IFN-gamma-producing cells (p=0.05) induced by casein stimulation in the group of 'non-allergic' children. Our data showed that immunologic parameters as IgE levels or CLA+ T cells in AD pediatric patients are influenced by the age, confirming that age could represent a bias in the analysis of immune response in those patients. Although, we demonstrated in children with AD the existence of different cytokine patterns of the lymphocyte response that could account for the different immunologic features between the two hypothesized forms of AD, which are not dependent on age. PMID- 15787868 TI - Is rhinitis alone or associated with atopic eczema a risk factor for severe asthma in children? AB - The objective of this study was to evaluate the role of rhinitis (R) and atopic eczema (E) on asthma severity among asthmatic (A) schoolchildren identified by the International Study of Asthma and Allergies in Childhood written questionnaire (WQ). WQ was applied to parents of 6-7-yr-old schoolchildren (SC, n=3033), and to adolescents (AD, 13-14 yr old, n=3487), living in Sao Paulo, Brazil. An affirmative response to 'has your child/have you had wheezing/whistling in the last year' identified those with A, and an affirmative response to 'the last 12 months has your child/have you had sneezing/runny/blocked nose when he/she you did not have a cold/flu?' identified those with R. Subjects with an affirmative response to 'has your child/have you had this itchy rash at any time in the past 12 months?' were identified as having E. Subjects who had R associated with A were identified as AR and those with A associated with R and E as ARE. A who had at least two affirmative responses to questions for asthma severity: speech disturbance, more than four acute attacks, sleep disturbance, and wheezing with exercise were defined as having severe asthma. 22.1% AD and 24.3% SC were identified as A; 47.1% of those AD and 42.0% SC had AR and 10.0% of those AD and 12.8% of SC had ARE. Considering ARE, AR and A groups, speech disturbance during an acute episode of asthma was significantly higher among ARE AD (20.0% vs. 11.5% vs. 8.7%, p<0.05), and ARE SC (22.1% vs. 13.9% vs. 10.5%, p<0.05) in comparison with A. Likewise, more than four acute attacks in the last year was significantly higher among ARE AD (24.0% vs. 14.0% vs. 10.5%, p<0.05) and ARE SC (32.6% vs. 19.4% vs. 12.8%, p<0.05) as the frequency of sleep disturbance due to wheezing, for AD (61.3% vs. 42.0% vs. 38.4%, p<0.05) and SC (77.9% vs. 67.3% vs. 58.4%, p<0.001) and for 'wheezing associated with exercise' for AD (72.0% vs. 47.5% vs. 39.9%, p<0.001) and SC (36.8% vs. 31.4% vs. 14.1%, p<0.001). Prevalence of severe asthma was higher among ARE AD (57.3% vs. 31.9% vs. 27.0%, p<0.05) and ARE SC (52.6% vs. 36.9% vs. 22.5%). In patients with A, the presence of R or E are risk factors for severe asthma, and both together (R and E) are a higher risk. PMID- 15787869 TI - Building characteristics affect the risk of allergy development. AB - Damp dwellings increase the risk for house dust mite (HDM) infestation in temperate climate zones and may be associated with an increased risk for allergic disease. The aim of the study was to assess possible relationships between allergen levels in house dust, characteristics of residence buildings and allergic diseases in children. A subsample of 12-yr-old children, having the same address in 1991 and 1996, was selected from a population-based sample of children from the Goteborg area. Health inspectors examined the residences of all the 109 children and several different building characteristics including humidity and indoor temperature were collected. Dust samples for analysis of HDM allergens were collected from the children's beds, and for analysis of cat and dog allergens from the living room. Current health status was assessed by questionnaires, interviews and skin prick tests (SPT). Dog or cat allergens were found in all houses, even in houses without such animals. HDM allergens were found in 60% of the houses, but only six of them had levels exceeding 2 microg/g dust. There was a strong association between HDM-infestation and wheeze, but not with specific sensitization to HDM. The type of building (houses when compared with flats), the ventilation system and the presence of a basement had all major implications on respiratory symptoms, atopy and HDM infestation. We can conclude that dog or cat allergens were found in all houses, and a strong association between HDM infestation and indoor environment. Building construction affected both respiratory morbidity and sensitisation independently, suggesting not only worsening of symptoms but also a causative relationship with disease development. PMID- 15787870 TI - Mites in dust samples from mattress surfaces from single beds or cribs in the south Brazilian city of Londrina. AB - The aim of this study was to investigate the mite fauna in mattresses dust samples from cribs or beds in the south Brazilian city of Londrina, State of Parana. A total of 133 dust samples from upper and lower mattress surfaces, and bed frames were aspirated once from 38 dwellings (18 cribs and 21 beds), and one day nursery (six cribs). A total of 758 mite bodies were counted in slides: 233 (30.7%) from cribs and 525 (69.3%) from beds (p<0.001). House dust mites--mainly Dermatophagoides pteronyssinus, represented 72% and 84% of total mite count in crib and bed dust samples, respectively. The mean HDM body concentration in crib or bed slides were, respectively, 289.9+/-136.7 and 875.0+/-183.6 mites/g. Statistical analysis showed a significantly higher mite bodies count on lower mattress surface compared with upper surface in bed samples only (p=0.025). Data herein show that cribs like mattress have sufficient mite bodies to cause sensitization to humans. The use of mattress covers for cribs and beds should be encouraged in order to avoid allergens exposure. PMID- 15787871 TI - Inverse association between Chlamydia pneumoniae respiratory tract infection and initiation of asthma or allergic rhinitis in children. AB - To evaluate the role of Chlamydia pneumoniae respiratory tract infection on pediatric asthma, allergic rhinitis or atopic eczema initiation, children of three age groups (n=1211) were prospectively studied for a C. pneumoniae infection using throat swabs and polymerase chain reaction (PCR) with enzyme immunoassay (EIA) detection. Infected children (study group, SG) were examined monthly until the agent could not be detected, quantifying persistent infection. They were compared with randomly selected, non-infected children without asthma matched for age, gender and origin (control group, CG) regarding lung function and inflammatory parameters as well as initiation of allergic diseases judged by family doctor diagnosis after, in median, 22 months. At the first follow-up examination, SG children revealed a higher leukotriene B4 (median 36 pg/ml vs. 19, p=0.04) and 8-isoprostane (median 15 pg/ml vs. 12, p=0.04) in breath condensate characterizing neutrophil, agent-related inflammation and oxidative stress in the lower airways. Cysteinyl leukotrienes, important in acute allergic inflammation, were without difference. Local, anti C. pneumoniae secretory immunoglobulin A antibodies were higher in children after C. pneumoniae infection (optical density median 0.7 vs. 0.4, p=0.001) confirming PCR-EIA results. At the final examination, there was no difference in pathological lung function tests, parameters of exhaled breath condensate or eosinophilia of the nasal mucosa. Incidence of asthma (0/55 vs. 5/54, p=0.03) and allergic rhinitis [3/53 vs. 10/52, p=0.04, odds ratio and 95% confidence interval-OR 0.25 (0.06;0.98)] as well as prevalence of asthma [1/56 vs. 9/58, p=0.02, OR 0.1 (0.01;0.81)] and allergic rhinitis [6/56 vs. 16/58, p=0.03, OR 0.32 (0.11;0.88)] were lower in the SG children. There was no association in atopic eczema. Three children with persistent infection revealed a slightly higher incidence in allergic rhinitis without significance than those with single C. pneumoniae detection (1/3 vs. 2/50), however, not to the CG. In conclusion a C. pneumoniae upper respiratory tract infection may be regarded as a protective factor for childhood asthma or allergic rhinitis in a population of kindergarten and school-age children. PMID- 15787872 TI - Why Chlamydia pneumoniae is associated with asthma and other chronic conditions? Suggestions from a survey in unselected 9 yr old schoolchildren. AB - Despite numerous studies demonstrating an association between asthma and many other chronic conditions and signs of Chlamydia pneumoniae (Cp) infection, the role of Cp in the pathogenesis of these illness remain still unclear. We investigated the prevalence of Cp antigen in the upper airways and the prevalence of detectable Cp serum antibodies in an unselected population of 207 9-yr-old schoolchildren. We also sought the presence of asthma, chronic or recurrent respiratory symptoms by means of questionnaire completed by the parents. Nasal aspirate, blood sampling and allergen skin prick tests were also performed. None of the children had obvious signs of acute infection at physical examination. Cp DNA was detected in nasal aspirates from 20 of the 207 children tested and serum IgG antibodies for Cp in 68 children. No association was found between atopy or history of atopic illness and the presence of Cp DNA or antibody production. This finding is explained by the fact that our study was conducted in an unselected childhood population, inherently including few children with asthma. A strong association between the status of antigen carrier and the presence of detectable Cp serum immunoglobulin (Ig)G or IgM suggests that subjects with detectable Cp antibodies have an impaired ability to eliminate this pathogen when infected. Because Cp eradication requires a strong Th1 lymphocyte response, the previously proven association between Cp and asthma, might reflect the known association of asthma with Th2-oriented lymphocytic activity. PMID- 15787873 TI - Prevalence of allergy-related symptoms in Singaporean children in the second year of life. AB - This study describes the cross-sectional prevalence of symptoms associated with eczema (chronic itchy rash), asthma (wheeze), and allergic rhinitis (rhinoconjunctivitis) in 1026 subjects between 18.5 and 23 months old (median age is 21 months) in Singapore. The first 2 yr cumulative prevalence of chronic itchy rash, wheeze, and rhinoconjunctivitis were 22.1% (n=227), 22.9% (n=235), and 8.4% (n=86) respectively. In total, 42.2% (414 of 979) reported ever having any of these symptoms. Eczema, although prevalent, was diagnosed only in 34.4% (n=78) of children with chronic itchy rash. Children with this eczematous rash were also more prone to wheeze (cOR=2.0, 95% CI: 1.2-3.0) and rhinoconjunctivitis (cOR=2.0, 95% CI: 1.4-2.8). Similarly, subjects who reported rhinoconjunctivitis and chronic itchy rash were 2.4 times (95% CI: 1.6-3.6) and 1.4 times (95% CI: 1.0 2.0) more at risk of wheezing respectively. Family history of allergy was a significant risk factor for chronic itchy rash (aOR=1.8, 95% CI: 1.3-2.4) and wheeze (aOR=1.7, 95% CI: 1.3-2.4). Thus, symptoms related to allergy were already prevalent during the second year of life. Significant proportions of these symptoms are likely to be due to true atopy as strong relationship with familial history and comorbidity with other potential allergic symptoms were observed. PMID- 15787874 TI - Changing prevalence of asthma in Taiwanese adolescents: two surveys 6 years apart. AB - This study compared the prevalence of asthma among Taiwanese adolescents with individual-level risk factors and municipal-level air pollution and meteorology data to determine whether changes in these factors could explain the observed change in prevalence. We conducted two national surveys of respiratory illness and symptoms in Taiwanese middle-school students in 1995-96 and 2001. The effects of personal and environmental factors were assessed and temporal changes of outdoor monitoring data were also compared with asthma prevalence difference. A total of 44,104 children from the 1995-96 survey and 11,048 children from the 2001 survey attended schools located within 1 km of 22 monitoring stations. Lifetime prevalences of physician-diagnosed and questionnaire-determined asthma increased during this period. After adjustment for potential risk factors, the prevalence differences were statistically unchanged. Although parental education level contributed most, changes in investigated personal and environmental factors might not explain the observed changes in asthma prevalence. Municipalities with higher temperature increase were significantly associated with prevalence difference in questionnaire-determined asthma. We concluded that correlates of the investigated individual-level factors, which have changed over time, still underlie changes in asthma prevalence. Increasing temperature might be the main reason for the rising trends of asthma in Taiwanese adolescents. PMID- 15787875 TI - Low prevalence of latex sensitivity in South African spina bifida children in Cape Town. AB - Spina bifida children have a high prevalence of latex allergy in studies reported from Europe and the USA. This study investigated the prevalence of latex allergy in a cohort of 24 spina bifida children at the Red Cross Children's Hospital from Cape Town, South Africa. The children were investigated using a detailed questionnaire, skin prick tests (ALK-Abello), ImmunoCap RASTs, Western blotting and ELISA, using the purified latex proteins Hev b1 and Hev b3 and whole latex preparation. A low overall prevalence of latex sensitization of 16.7% was found in the children. Children who were sensitive reacted to water insoluble to Hev b1 and Hev b3 proteins. The low prevalence of latex sensitization in the South African children may not be entirely explained by stringent latex avoidance. The children were from a low socioeconomic social status and 'hygiene' and other factors should be considered. PMID- 15787876 TI - Sesame allergy in Britain: a questionnaire survey of members of the Anaphylaxis Campaign. AB - Sesame is a major allergen in countries where it is a common food. It was noted that an increasing number of members of the UK charity, the anaphylaxis campaign, were reporting allergy to sesame. This study, sought to examine features of sesame allergy among members of the Anaphylaxis Campaign (which supports those at potentially life-threatening risk from allergies) focusing on clinical symptoms and features of the foods implicated. A physician-designed questionnaire was sent by post to 400 members of the Anaphylaxis Campaign who reported avoidance of sesame. Two hundred and eighty replies were received (70%). Twenty-three replies (7%) were excluded and 96 replies (24%) came from subjects who avoided sesame but had never reacted to it. One hundred and fifty people (54%) reported 288 reactions to sesame. 89% of reactive subjects reported other atopic diseases and notably 84% were also nut/peanut allergic. One in six (17%) had suffered potentially life-threatening symptoms, with 65% of severe reactions happening on first known exposure. The age of first reaction ranged from 6 months to 65 yr. The majority of reactions reported (91%) involved foods or dishes which had sesame as a deliberate ingredient, rather than sesame as an accidental contaminant. Respondents represented a well-informed and highly selected group of people at risk from potentially life-threatening allergies. Sesame should be identified clearly as an ingredient and separately from nuts and peanuts when it may be an allergen contaminant. People at potential risk need clear allergy diagnosis and informed guidance to enable them to avoid this key allergen more easily. PMID- 15787877 TI - Agreement between parental and self-completed questionnaires about asthma in teenagers. AB - In studies of asthma in children, a common method is for the parents to complete questionnaires about their child's asthma symptoms. With longitudinal studies of asthma, children reach an age when they can complete the questionnaire themselves. The aim of this paper was to compare the prevalence of asthma symptoms as well as the agreement between responses to an asthma questionnaire completed by teenagers and their parents. As a part of the Obstructive Lung Disease in Northern Sweden Study (OLIN) pediatric study, where 3345, 13-14-yr-old children completed an asthma questionnaire, 294 (84%) randomly selected parents also completed the questionnaire, which included the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire. There were no significant differences in the prevalence of wheeze in the last 12 months, ever asthma, or physician diagnosed asthma as reported by the parents compared with the teenagers. However, the teenagers reported a significantly higher prevalence of wheeze during or after exercise. The absolute agreement was generally very high while the level of agreement (kappa-value) was slightly lower. The highest results in both absolute agreement and kappa-value, were reached by the questions on diagnosis of asthma (98.9% and 0.93), use of asthma medicines (95.5% and 0.78), and whether the child ever had had asthma (97.2% and 0.86), respectively. In conclusion, the agreement between the parents' and the teenagers' responses to the asthma questionnaire was good. The change in methodology from parental to self-completion of the questionnaire did not affect the results in the study. PMID- 15787878 TI - Computer aided design mapping for SCORAD index in atopic dermatitis--accessible and economical. PMID- 15787879 TI - Are intranasal steroids effective in the treatment of 2-4-year-old children with perennial rhinitis? PMID- 15787880 TI - Bill Silverman: a personal appreciation. PMID- 15787881 TI - Mandatory rescue of fetal infants. PMID- 15787882 TI - Smoking during pregnancy and preterm birth according to obstetric history: French national perinatal surveys. AB - The relation between smoking and preterm delivery is not totally known. Our aim was to determine whether smoking during pregnancy was associated with preterm birth among women at different risk according to their obstetric history. The study was based on data from the 1998 French national perinatal survey. Of the 13073 singleton live births, 4.7% were preterm; 15% of the pregnant women were moderate (one to nine cigarettes per day) and 10% heavy smokers (at least 10 cigarettes per day). Smoking heavily was related to preterm birth (crude odds ratio [OR] = 1.35, 95% confidence interval [95% CI]: [1.04, 1.74]). Multivariable logistic regression showed a relation between smoking and preterm birth among multiparae without previous adverse pregnancy outcomes; the associated adjusted ORs (AORs) were 1.25 [95% CI 0.83, 1.87] among moderate smokers and 1.46 [95% CI 0.98, 2.20] among heavy smokers. The corresponding AORs were 0.69 [95% CI 0.46, 1.05]) and 0.96 [95% CI 0.59, 1.56] for primiparae and 1.11 [95% CI 0.63, 1.93] and 0.50 [95% CI 0.25, 0.98] for multiparae with previous adverse pregnancy outcomes. Our study showed a relation between heavy smoking during pregnancy and preterm birth mostly for women with low obstetric risk. PMID- 15787884 TI - Should spontaneous and medically indicated preterm births be separated for studying aetiology? AB - An increasing proportion of preterm births result from medical interventions, and the practice of aggregating all preterm births vs. splitting into spontaneous and medically indicated preterm births is inconsistent. While mechanistic and clinical arguments can be offered for either approach, we empirically evaluated the predictiveness of a range of risk factors for preterm birth in the Pregnancy, Infection, and Nutrition Study. Most influences were shared across the two subsets - African-American ethnicity, advancing age, delivery at a university medical centre, prior preterm birth and smoking. Medically indicated preterm births appeared to be associated with intensity of medical care, higher in the university medical centre and lower for the poorest women. Body mass index was positively associated with medically indicated preterm birth and inversely with spontaneous preterm birth. Given the complexity of the aetiological pathways, both aggregation and disaggregation are well justified and should be included in studies of the causes of preterm birth. PMID- 15787885 TI - Short-term paracetamol therapy during pregnancy and a lower rate of preterm birth. AB - The relationships between short-term oral paracetamol treatment during pregnancy and mean gestational age and birthweight, in addition to the prevalence of preterm birth and low birthweight were estimated using newborn infants in the population-based large dataset of the Hungarian Case-Control Surveillance of Congenital Abnormalities between 1980 and 1996. Gestational age and birthweight were medically recorded. Of 38 151 newborn infants, 173 were born to mothers who had paracetamol treatment during pregnancy. A 0.4-week increase in the mean gestational age was found among exposed relative to unexposed, and with the use of paracetamol during pregnancy there was a significant reduction in the proportion of preterm birth (3.5% vs. 9.2%) (adjusted P = 0.01). The possible preterm birth protective effect of paracetamol may be associated with paracetamol induced reduction of prostacyclin production in pregnant women. The clinical relevance of these findings needs further studies. PMID- 15787886 TI - Folic acid supplements in pregnancy and birth outcome: re-analysis of a large randomised controlled trial and update of Cochrane review. AB - Periconceptual folic acid prevents neural tube defects. The effect of folic acid taken throughout pregnancy is unclear, however. We re-analysed data from a large randomised controlled trial performed between 1966 and 1967 and combined the results with those from trials included in a Cochrane review. A total of 2928 women were randomised: 1977 were allocated to placebo, 466 to folic acid 200 microg/day and 485 to folic acid 5 mg/day. Folic acid supplementation was not associated with any difference in mean birthweight, placental weight or gestational age. When combined with trials in the Cochrane review folic acid at high doses was associated with reduced risk of low birthweight (pooled relative risk 0.73 [95% CI 0.53, 0.99]). We found no conclusive evidence of benefit for folic acid supplementation in pregnant women given from time of booking onwards. PMID- 15787887 TI - The association of time in the US and diet during pregnancy in low-income women of Mexican descent. AB - This study compared nutritional intake during pregnancy among women of Mexican descent according to country of birth (US vs. Mexico) and, for Mexico-born women, according to number of years lived in the US (or= 11 years). A 72-item food frequency questionnaire (FFQ) was used to assess dietary intake in 474 pregnant Mexico-born immigrants and US-born Mexican-Americans. Mexico-born women had significantly higher intakes of calories (P = 0.02), fibre (P < 0.001), vitamin A (P < 0.001), vitamin C (P = 0.03), vitamin E (P < 0.01), folate (P < 0.01), calcium (P < 0.001) and zinc (P = 0.02) from their diets than US-born women. Intakes of all nutrients except vitamin C and zinc remained significantly higher in Mexico-born women when nutrients from both diet and vitamin supplements were considered. Among Mexico-born women, increasing years of residence in the US was associated with lower intake of calories (P(trend) < 0.01), fibre (P(trend) < 0.01), folate (P(trend) = 0.03), iron (P(trend) = 0.05) and zinc (P(trend) = 0.03), although only the trend for iron remained significant when vitamin supplement sources were included. A large percentage of women had inadequate intake of vitamin E (58%), folate (61%), iron (77%) and zinc (47%) from their diets during pregnancy and these rates were higher in US-born women than Mexico-born women. PMID- 15787888 TI - Ethnic differences in congenital malformations in the Netherlands: analyses of a 5-year birth cohort. AB - Congenital malformations are among the major causes of perinatal mortality and morbidity at present. Research into the ethnic diversity of congenital malformations can form a basis both for aetiological studies and for health care advice and planning. This study compared the overall prevalence of congenital malformations, the prevalence in different organ systems and of several specific malformations between different maternal ethnic groups in the Netherlands using a 5-year national birth cohort (1996-2000) containing 881 800 births. Maternal ethnic groups considered were Dutch; Mediterranean (Moroccan/Turkish); other European; Black; Hindu and Asian. Mediterranean women had a 20% higher risk of having a child with a congenital malformation than Dutch women (age-adjusted OR = 1.21 [95% CI 1.16, 1.27]). They showed an increased risk of malformations in several organ systems such as the central nervous system and sensory organs, the urogenital system and skin and abdominal wall. Further, they had an increased risk of the group of chromosomal malformations/multiple malformations/syndromes. For the specific group of multiple malformations the maternal age adjusted OR was 1.80 [95% CI 1.47, 2.20]. The Black group showed a significantly increased risk of skeletal and muscular malformations (age adjusted OR = 1.76 [95% CI 1.53, 2.02]) with a sixfold increased risk of polydactyly compared with the Dutch group. For Mediterranean women, the largest and fastest growing group of immigrants in the Netherlands, this study demonstrated an increased risk of congenital malformations. PMID- 15787889 TI - Infant mortality in Israel during 1950-2000: rates, causes, demographic characteristics and trends. AB - We evaluated the trends and risk factors in infant mortality in Israel over five decades (1950-2000), based on data obtained from the official notifications of live births, and death certificates. Until the 1960s the main cause of infant mortality was infectious disease; this was replaced by congenital anomalies in Moslems and Druzes, and preterm birth in Jews and Christians. In 2000, there were 746 infant deaths, and the national infant mortality rate (IMR) was 5.4 per 1000 live births (Jews 3.9; [95% CI 3.5, 4.3]; Moslems 9.2 [8.3, 10.3]; Christians 3.6 [1.4, 5.8]; Druzes 6.3 [3.6, 9.0]). Between 1955 and 2000 the overall IMR declined sevenfold (absolute declines of 56.8, 56.3, 45.0 and 28.3 per 1000 live births, in Moslems, Druzes, Christians and Jews, respectively). The reduction in IMRs between 1990 and 2000 in all religious groups (>45%) exceeded the goal set by the World Summit for Children in 1990 of 33%. In 2000, the main risk factors were birthweight < 1500 g [relative risk (RR) = 69], major congenital malformations (RR = 22.0 [18.8, 25.7], and multiple births (RR of 9.3 and 4.2 in triplets and twins respectively). We conclude that the marked decline in IMRs in Israel over five decades reflects a major improvement in population health. Today, infant mortality in Israel represents a unique combination of high rate of congenital malformations among Moslems, where consanguineous marriages are common, and medical termination of pregnancy of malformed fetuses are infrequent; and relatively high IMRs from preterm birth in Jews, associated with high rates of assisted reproduction. PMID- 15787890 TI - Allergy and infectious disease histories and the risk of childhood acute lymphoblastic leukaemia. AB - Infectious disease histories were evaluated in a population-based case-control study of childhood acute lymphoblastic leukaemia (ALL) as it has been hypothesised that delays in early infections are associated with an increased risk of disease. Allergy histories were also assessed as part of a broader evaluation of the role of immune factors in ALL. Cases (n = 255) were diagnosed between 1980 and 1991 at one of four referral centres in a 31-county area of New York State; controls (n = 760) were a random sample of live births from the same region, frequency matched to cases by sex, race and birth year. Data were collected by mailed questionnaire, completed by case and control parents in 1995. Allergy and infectious histories before the age at leukaemia diagnosis for cases and an equivalent age for controls were evaluated. The adjusted odds ratio and 95% confidence interval [CI] associated with a positive history of any allergy was 0.58 [95% CI 0.38, 0.88] compared with a negative allergy history. The occurrence of several common childhood illnesses before 25 months of age and ALL were assessed, with both weak positive and weak inverse associations observed. Overall, these analyses provide little support for the hypothesis that infection delay in early life is associated with an increased risk of ALL. Children with positive allergy histories reported significantly more infections than those with negative histories; however, effect modification of the infection-ALL associations by child allergy history was not observed. Nonetheless, these observations suggest the importance of assessing both allergy and infectious histories and their possible interactions when evaluating the association between these immune factors and childhood ALL. PMID- 15787891 TI - Multilevel modelling of hospitalisations for recurrent diarrhoeal disease in Aboriginal and non-Aboriginal infants and young children in Western Australia. AB - A multilevel proportional hazards model was used to determine the prognostic factors affecting hospitalisations for recurrent diarrhoeal disease in infants and young children. All infants born in 1996 who had index gastroenteritis admission to Western Australian (WA) hospitals during their first year of life were included in our study cohort (n = 486). There were 618 hospital admissions for infectious diarrhoea over the 6-year follow-up period. Aboriginal children stayed significantly longer in hospital than their non-Aboriginal peers, and comorbidities such as dehydration, gastrointestinal sugar intolerance, failure to thrive, iron deficiency anaemia and certain infections (genitourinary, scabies and/or otitis media) were all significantly associated with the delayed discharge of patients. Substantial variations among patients (variance = 0.660) and between health service regions of WA (variance = 0.296) were found. Over 30% of the total variation could be attributed to the heterogeneity between health districts. For any two patients in the same health district, the within-region (intraclass) correlation was estimated to be 0.309. In the absence of detailed socio demographic data, application of the standard survival procedure may lead to incorrect inferences due to regional clustering and repeated observations on individuals. By accounting for latent patient and regional effects, the multilevel analysis clearly confirmed the high burden of infectious diarrhoea among Aboriginal infants and children, and their much longer hospital stays. Coexisting morbidities contributed to the prolonged and recurrent hospitalisations. Findings of this epidemiological study indicated the need of multifaceted clinical disease prevention and hygiene promotion strategies to control the disease. PMID- 15787892 TI - From caution to certainty: hazards in the formation of evidence-based practice - a case study on evidence for an association between the use of uterine stimulant drugs and amniotic fluid embolism. PMID- 15787894 TI - Insulin resistance in children and adolescents with type 1 diabetes mellitus: relation to obesity. PMID- 15787895 TI - Insulin resistance in children and adolescents with type 1 diabetes mellitus: relation to obesity. AB - BACKGROUND: Insulin resistance is well recognized both in type 1 diabetes mellitus (T1DM) and in obesity. Studies concerning the relation between insulin resistance and overweight in T1DM have not yet been carried out. METHODS: Degree of overweight [standard deviation score-body mass index (SDS-BMI)] and daily insulin doses per weight (ID/kg), per body surface (ID/m2), and per ideal body weight (ID/IW) were recorded in 4124 children aged between 5 and 20 yr with a duration of T1DM of 4-5 yr and an adequate metabolic control [hemoglobin A1c (HbA1c) <8.0%]. SDS-BMI was compared between insulin-resistant (ID/kg > or = 1.0) and insulin-sensitive (ID/kg <1.0) children. The ID/kg, ID/m2, and ID/IW were compared between obese (SDS-BMI >1.9) and non-obese children. Multivariate linear regression analysis was conducted for the dependent variables ID/kg, ID/m2, and ID/IW, including age, gender, SDS-BMI, and HbA1c as independent variables. RESULTS: The 882 insulin-resistant children did not differ significantly (p = 0.447) with respect to SDS-BMI (median +0.38) compared to the 3242 insulin sensitive children (median SDS-BMI +0.42). The ID/kg was significantly (p = 0.031) lower in the obese children compared to the non-obese children (median 0.80 vs. 0.83), while ID/m2 (median 31.0 vs. 26.2) and ID/IW (median 1.17 vs. 0.85) were significantly (p < 0.001) increased in the obese children. In multivariate linear regression analysis, SDS-BMI was significantly (p < 0.001) associated with an increase in ID/m2 and ID/IW and a decrease in ID/kg. CONCLUSIONS: T1DM children with insulin resistance based on ID/kg are not more overweight than insulin-sensitive children with T1DM. ID/m2 and ID/IW seem to reflect a better tool than ID/kg to describe the influence of overweight on insulin resistance in T1DM. PMID- 15787896 TI - Comparison of fingerstick hemoglobin A1c levels assayed by DCA 2000 with the DCCT/EDIC central laboratory assay: results of a Diabetes Research in Children Network (DirecNet) Study. AB - BACKGROUND: The Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications (DCCT/EDIC) high-performance liquid chromatography (HPLC) method for measuring hemoglobin A1c (HbA1c) serves as a reference standard against which other assays are compared. The DCA 2000 + Analyzer (Bayer Inc., Tarrytown, NY, USA), which uses an immunoassay, is a very popular device for measuring HbA1c levels in pediatric diabetes practices. OBJECTIVE: To determine how HbA1c values measured with the DCA 2000 in a multisite, pediatric diabetes clinic setting compare with corresponding HbA1c values measured in the DCCT/EDIC laboratory. DESIGN/METHODS: To examine this question, the Diabetes Research in Children Network (DirecNet) used the DCA 2000 in five clinical centers to measure baseline HbA1c levels in 200 youth with type 1 diabetes mellitus (T1DM) (aged 12.5 +/- 2.8 yr) who were participating in an outpatient clinical trial. At the same visit, an additional blood sample was obtained, refrigerated, and shipped to the DCCT/EDIC central laboratory for determination of HbA1c values. RESULTS: The central laboratory HbA1c value averaged 8.0 +/- 0.9% (mean +/- SD), with a median (25th and 75th quartiles) of 7.8% (7.3 and 8.5%, respectively). The DCA 2000 HbA1c values were strongly correlated (r = 0.94, p < 0.001), but significantly higher than DCCT/EDIC central laboratory values with a mean difference of +0.2% (95% confidence interval +0.14 to 0.23%, p < 0.001). There was some variation in the differences between DCA 2000 and central laboratory values at the five clinical centers (p < 0.001) with mean differences ranging between 0.0 and 0.3%, but differences between the two methods did not vary significantly by age or gender. CONCLUSION: Measurements of HbA1c by the DCA 2000 compare favorably with the DCCT/EDIC central laboratory method, albeit with slightly higher values. PMID- 15787897 TI - Continuous subcutaneous insulin infusion in toddlers and children with type 1 diabetes mellitus is safe and effective. AB - OBJECTIVE: The aim of this study was to investigate the safety and efficacy of continuous subcutaneous insulin infusion (CSII) for type 1 diabetes mellitus (T1DM) in toddlers and children. RESEARCH DESIGN AND METHODS: Seventy children who began CSII at the age of 12 yr or younger (youngest 2 yr old) and who were maintained on CSII for at least 6 months were studied by a retrospective chart review. A pre- or postintervention comparison approach was used to assess the impact of CSII on the measured variables. The control period was defined as 1 yr prior to beginning CSII. Charts were reviewed for hemoglobin A1c (HbA1c) reports of severe hypoglycemia, diabetic ketoacidosis (DKA), height and weight, and range of blood glucoses reported at each visit. Mean values for HbA1c, body mass index (BMI) z-score, and range of blood glucose were computed for each subject over all pre-CSII visits, and again over all post-CSII visits. RESULTS: The mean HbA1c decreased significantly during CSII [7.8 +/- 0.8% pre-CSII vs. 7.3 +/- 0.7% on CSII, p < 0.0001]. Hypoglycemic episodes decreased with CSII in the 10- to 12-yr old group (p < 0.02) and demonstrated a strong trend (mean of 0.46-0.22 events per patient year, p < 0.06) overall. Two episodes of DKA occurred in the CSII period and none in the control period (p = NS). BMI z-scores increased to 0.21 in the 5- to 9-yr-age group (p < 0.008) and averaged 0.13 overall. The range of blood glucoses decreased during CSII (p < 0.005) in the middle and oldest age groups. CONCLUSIONS: This study supports CSII as a safe and effective alternative to managing T1DM, with no increase in hypoglycemia and a trend to improve control, even in the youngest patients. PMID- 15787898 TI - Insulin pump use in young adolescents with type 1 diabetes: a descriptive study. AB - There is little research on psychosocial factors and insulin pump use in adolescents. The purpose of the present study was to use qualitative and quantitative methods to explore psychosocial issues related to insulin pump use [continuous subcutaneous insulin infusion (CSII)] in youth. Eighteen early adolescents and their parents were interviewed about the experience of using an insulin pump, and transcripts were content-analyzed. In general, interviewees reported modest improvements in glycemic control with initiation of CSII. Teens and parents did, however, report high levels of satisfaction with pump therapy and increased adolescent responsibility for the diabetes regimen. In addition, pump users reported few issues related to body image, appearance, or social aspects of pump dependency. Chief concerns were related to the demands of initiating pump therapy, pump alarms and malfunctions, potential for regimen non compliance with CSII, and school-related issues. Pump use is not associated with social difficulties, and, in general, youths and their parents report high satisfaction with CSII. PMID- 15787900 TI - Cerebral edema in diabetic ketoacidosis and other acute devastating complications: recent observations. PMID- 15787899 TI - A randomized trial of regular standardized telephone contact by a diabetes nurse educator in adolescents with poor diabetes control. AB - OBJECTIVE: The aim of this study was to determine the effect of regular standardized telephone contact by a diabetes nurse educator (DNE) on metabolic control, treatment compliance, and quality of life in adolescents with poorly controlled type 1 diabetes. METHODS: A single-blinded 6-month randomized controlled trial was used. Participants included 46 of 49 eligible adolescents (13-17 yr) with type 1 diabetes >1-yr duration and hemoglobin A1c (HbA1c) >8.5% for the previous 6 months. Subjects were randomly assigned to 6 months of standard diabetes management or standard care plus weekly telephone contact by a DNE. Telephone conversations included review of events in the adolescents' lives and diabetes education, but the primary focus was on blood glucose results and insulin-dose adjustments. HbA1c, compliance with glucose monitoring, quality of life [Diabetes Quality of Life Scale for Youth (DQOLY)], and family functioning [Family Environment Scale (FES)] were assessed at baseline, and at 3 and 6 months. Posthoc, HbA1c levels were assessed 6 months following study completion. RESULTS: Six months of regular telephone contact by a DNE had no immediate effect on any of the outcome measures. However, posthoc 6 months, HbA1c levels decreased (1% change compared to baseline) in 6/21 of the study group and 0/18 of the control group, while HbA1c increased in 4/21 of study subjects compared to 8/18 of control subjects (p = 0.015). CONCLUSIONS: In contrast to adult studies, regular telephone contact did not lead to immediate improvements in metabolic control in adolescents with poorly controlled type 1 diabetes. However, knowledge and skills gained during the intervention may have had a delayed beneficial effect in these high-risk adolescents. PMID- 15787901 TI - Self-monitoring of blood glucose in children and teens with diabetes. AB - Improved metabolic control has unequivocally been demonstrated to delay the onset and slow the progression of microvascular complications in adolescents and adults with diabetes mellitus. Growing evidence also supports the association of tighter glucose control and more frequent blood glucose monitoring. Therefore, self monitoring of blood glucose (SMBG) has become a fundamental part of diabetes care in children. Here, we review recent advances and ongoing trends in glucose monitoring in children with diabetes. Technologies have been developed to improve patient compliance with recommended monitoring, requiring less blood, involving less pain, and providing results more quickly. Alternate-site testing (AST) is also a potential means of improving patient compliance with SMBG by avoiding the sensitive fingertip area. The Continuous Glucose Monitoring System (CGMS) and the GlucoWatch Biographer are two recent tools that can track glucose levels continuously. However, inconsistency in their accuracy and precision remain challenges when using these technologies to guide management. PMID- 15787903 TI - On the fact-value distinction and the phenomenology of caring. PMID- 15787904 TI - Narrative vigilance: the analysis of stories in health care. AB - The idea of narrative has been widely discussed in the recent health care literature, including nursing, and has been portrayed as a resource for both clinical work and research studies. However, the use of the term 'narrative' is inconsistent, and various assumptions are made about the nature (and functions) of narrative: narrative as a naive account of events; narrative as the source of 'subjective truth'; narrative as intrinsically fictional; and narrative as a mode of explanation. All these assumptions have left their mark on the nursing literature, and all of them (in our view) are misconceived. Here, we argue that a failure to distinguish between 'narrative' and 'story' is partly responsible for these misconceptions, and we offer an analysis that shows why the distinction between them is essential. In doing so, we borrow the concept of 'narrativity' from literary criticism. Narrativity is something that a text has degrees of, and our proposal is that the elements of narrativity can be 'sorted' roughly into a continuum, at the 'high narrativity' end of which we find 'story'. On our account, 'story' is an interweaving of plot and character, whose organization is designed to elicit a certain emotional response from the reader, while 'narrative' refers to the sequence of events and the (claimed) causal connections between them. We suggest that it is important not to confuse the emotional persuasiveness of the 'story' with the objective accuracy of the 'narrative', and to this end we recommend what might be called 'narrative vigilance'. There is nothing intrinsically authentic, or sacrosanct, or emancipatory, or paradigmatic about narrative itself, even though the recent health care literature has had a marked tendency to romanticize it. PMID- 15787905 TI - Dangerous and severe personality disorder: an ethical concept? AB - Most clinicians and mental health practitioners are reluctant to work with people with dangerous and severe personality disorders because they believe there is nothing that mental health services can offer. Dangerous and severe personality disorder also signals a diagnosis which is problematic morally. Moral philosophy has not found an adequate way of dealing with personality disorders. This paper explores the question: What makes a person morally responsible for his actions and what is a legitimate mitigating factor? How do psychiatric nurses working with this client group understand the awful things some clients do? What concepts do they need, if they are to know how to explain and how to react? It is suggested that dangerous and severe personality disorder is best regarded as a moral category, framed in terms of goodness, badness, obligation and other ethical concepts. It seems plausible that in important ways the dangerous and severe personality disordered client does not understand morality or understands it differently. The peculiar position of the dangerous and severe personality disordered individual in our system of moral responsibility stems from his apparent inability to see the importance of the interests of others. It might be more helpful to regard personality disordered clients as we do children: partially but not fully reasonable for their actions. We might regard the dangerous and severe personality disordered client responsible for those actions which he most clearly understands, such as causing others physical pain, but not for those with which he is only superficially engaged, such as causing emotional pain. The paper concludes by suggesting that the dangerous and severe personality disordered individual does not fit easily into any conventional moral category, be it criminal, patient, animal or child, and thus an assessment of his moral accountability must take into consideration his special circumstances. PMID- 15787906 TI - Narrative ethics in nursing for persons with intellectual disabilities. AB - Both in The Netherlands and in Britain, practices of 'life story work' have emerged in nursing for persons with intellectual disabilities. The narrative approach to care and support may at the same time be considered as an attempt to compensate for the 'disabled authorship' of many persons with intellectual disabilities and as a sign of controversy with standard practices of diagnosis and treatment that tend to neglect the personal identities of both clients and care givers, their particular historical and relational contexts and their spiritual needs. This paper argues that narrative ethics not only offers an appropriate moral framework for practices of life story work, but that these practices are a narrative ethics in action. Starting with an account of the concept of 'life story work' as it has been introduced in nursing practices in the field of intellectual disability, the paper explains its relationship with key characteristics of narrative ethics. The teleological dimension in narrative ethics and in practices of life story work sparks off a dialectic process of understanding of the client and self-understanding of the care giver. It also invites a respect for life in its openness toward the future and presupposes an openness toward other possible versions of the life narrative. The phenomenological and hermeneutic-interpretative methodologies in narrative ethics aim at a 'sudden moment of intimacy' in relationships of nurses and clients. The 'epiphany' of this essential moment of recognition, insight and engagement cannot, however, be brought about by methodology. PMID- 15787907 TI - The shifting sands of self: a framework for the experience of self in addiction. AB - The self is a common yet unclear theme in addiction studies. William James's model of self provides a framework to explore the experience of self. His model details the subjective and objective constituents, the sense of self-continuity through time, and the ephemeral and plural nature of the changing self. This exploration yields insights into the self that can be usefully applied to subjective experiences with psychoactive drugs of addiction. Results of this application add depth to the common understanding of self in addiction, acknowledge the importance of feelings and choice in the sense of self created in addiction experiences, and affirm the values salient to these interior experiences in addiction. These results suggest meaning derived from those values, and provide important background knowledge for the nurse interacting with these clients. PMID- 15787908 TI - Can there be a 'cosmetic' psychopharmacology? Prozac unplugged: the search for an ontologically distinct cosmetic psychopharmacology. AB - 'Cosmetic psychopharmacology' is a term coined by Peter Kramer in his 1993 best seller, Listening to Prozac. It has come to refer to the use of psychoactive substances to effect changes in function for conditions that are either normal or subclinical variants. In this paper, I ask: What distinguishes an existential ailment from clinical depression, or either of those from normal depressed mood, melancholic temperament, dysthymia or other depressive disorders? Can we reliably distinguish one from the other? Are the boundaries of illness and disorder really so distinct? If not, how can we know that treatment of 'depression' with Prozac in any given instance constitutes a cosmetic as opposed to, say, a medical or clinical use of psychopharmacology - a distinction that seems to turn on our ability to clearly differentiate the clinical from the cosmetic. If we cannot reliably distinguish between such conditions, can we even have a cosmetic psychopharmacology that is not a form of malpractice, broadly speaking? What if we unplugged Prozac from all the amplitude and hype that resulted in Listening to Prozac becoming an instant best-seller and simply asked whether or not we can clearly distinguish an appropriate cosmetic use of Prozac for 'depression' from an inappropriate cosmetic use of Prozac, and both of those from Prozac's appropriate clinical, that is, non-cosmetic uses? If we cannot make these distinctions, perhaps it is too early to say there can be such a thing as a cosmetic psychopharmacology. PMID- 15787909 TI - Plato's cave and Aristotle's collections: dialogue across disciplines. PMID- 15787910 TI - Human death. PMID- 15787915 TI - Midazolam as premedication: is the emperor naked or just half-dressed? PMID- 15787916 TI - Sevoflurane and epileptiform EEG changes. AB - Sevoflurane has become the volatile agent of choice for inhalation induction of anesthesia. Hemodynamic stability and lack of respiratory irritation have justified its rapid extension to pediatric inhalation induction. The epileptogenic potential of sevoflurane has been suspected since the first case reports of abnormal movements in children without a history of epilepsy. The objectives of this short review are to: (i) analyze clinical and electroencephalographic (EEG) features supporting epileptogenic activity of sevoflurane, (ii) identify factors which may modulate that activity, and (iii) suggest guidelines of clinical practice to limit expression of this epileptiform phenomenon, which has thus far unknown morbidity. The use of sevoflurane may be associated with cortical epileptiform EEG signs, usually without clinical manifestation. No lasting neurological or EEG sequelae have been described thus far, and the potential morbidity of this epileptogenic effect is unknown. The use of sevoflurane in children, with its remarkable cardiovascular profile, should include a number of precautions. Among them, the limitation of the depth of anesthesia is essential. The wide use of cerebral function monitoring (the most simple being the EEG), may permit optimization of sevoflurane dose and avoidance of burst suppression and major epileptiform signs in fragile subjects, notably the very young and the very old. PMID- 15787917 TI - Premedication in children: hypnosis versus midazolam. AB - BACKGROUND: The main objectives of premedication in children are to facilitate the separation from the parents, to reduce preoperative anxiety, to smooth the induction of anesthesia and to lower the risk of postoperative behavioral disorders. The most common technique is sedative premedication with midazolam. Hypnosis enables a state of relaxation to be achieved and has never been evaluated as a premedication technique. The aim of the present study was to evaluate the efficacy of hypnosis on anxiety and perioperative behavioral disorders versus midazolam. METHODS: Fifty children from 2 to 11 years of age were randomized into two groups: group H received hypnosis as premedication; group M were given 0.5 mg x kg(-1) midazolam orally, 30 min before surgery. Preoperative anxiety was evaluated using the Modified Yale Preoperative Anxiety Scale (mYPAS) score when arriving in the department (T1), when entering the operating room (T2), and when fitting the facemask (T3). Postoperative behavioral disorders were evaluated using the Posthospitalization Behavioral Questionnaire (PHBQ) at days 1, 7 and 14. RESULTS: The two groups showed no significant difference preoperatively with the PHBQ: (M) 21 (17-25) vs (H) 20 (8-25) and mYPAS score: (M) 28 (23-75) vs (H) 23 (23-78). The number of anxious children was less during induction of anesthesia in the hypnosis group (T3: 39% vs 68%) (P < 0.05). Postoperatively, hypnosis reduced the frequency of behavior disorders approximately by half on day 1 (30% vs 62%) and day 7 (26% vs 59%). CONCLUSIONS: Hypnosis seems effective as premedication in children scheduled for surgery. It alleviates preoperative anxiety, especially during induction of anesthesia and reduces behavioral disorders during the first postoperative week. PMID- 15787918 TI - Pediatric intravenous paracetamol (propacetamol) pharmacokinetics: a population analysis. AB - BACKGROUND: The aim of this study was to describe propacetamol pharmacokinetics in children in order to predict concentrations after a standard dosing regimen of propacetamol 30 mg x kg(-1) (15 mg x kg(-1) paracetamol) 6 h. METHODS: A population pharmacokinetic analysis of paracetamol time-concentration profiles (846 observations) from 144 children [postconception age (PCA) 27 weeks-14 years] was undertaken using nonlinear mixed effects models (NONMEM). These data were taken from seven separate studies involving children given intravenous propacetamol. Time-concentration profiles (503 observations) from a further 86 children (PCA: 37 weeks-14 years) given paracetamol elixir orally were included in the analysis to assess relative bioavailability of intravenous propacetamol. RESULTS: A three-compartment (depot, central and peripheral) linear disposition model fitted data better than a two-compartment (depot and central) model. Population parameter estimates (between subject variability, %) were central volume (V2/F(oral)) 24 (55%) l x 70 kg(-1), peripheral volume of distribution (V3/F(oral)) 30 (32%) l x 70 kg(-1), clearance (CL/F(oral)) 16 (40%) l x h(-1) x 70 kg(-1) and intercompartment clearance (Q/F(oral)) 55 (116%) l x h(-1) x 70 kg( 1). Clearance increased from 27 weeks PCA (1.87 l x h(-1) 70 kg(-1)) to reach 84% of the mature value by 1 year of age (standardized to a 70 kg person using allometric '1/4 power' models). Peripheral volume of distribution decreased from 27 weeks PCA (45.0 l x 70 kg(-1)) to reach 110% of its mature value by 6 months of age. Central volume of distribution and intercompartment clearance did not change with age. Between occasions variability for the peripheral volume of distribution (V3/F(oral)) and clearance (CL/F(oral)) were 18.5 and 19.3%, respectively. A rate constant representing hydrolysis of propacetamol to paracetamol (K(a) 96 h(-1)) was size related, but not age related. The relative bioavailability of intravenous propacetamol compared with an oral elixir was 0.5. CONCLUSIONS: A mean paracetamol serum concentration of 10 mg x l(-1) is achieved in children 2-15 years given a standard dose of propacetamol 30 mg x kg(-1) 6 h. This concentration in the effect compartment is associated with a pain reduction of 2.6/10 after tonsillectomy and provides satisfactory analgesia for mild to moderate pain. Clearance is reduced in children less than 1 year of age and the target concentration of 10 mg x l(-1) may be achieved by scaling this standard dose regimen using predicted clearance in this younger age group. PMID- 15787919 TI - Syringe pumps take too long to give occlusion alarm. AB - BACKGROUND: The aim of this study was to assess the time needed to trigger an occlusion alarm, and the influence of the type of infusion pump, type and size of the syringe, and the set infusion rate. METHODS: Forty syringe pumps (20 JMS SP 100 and 20 JMS SP-500 machines) were tested using two types (JMS and Hayat) and two sizes (20 and 50 ml) of syringes at four infusion rates (0.5, 1, 2 and 5 ml x h(-1)). Syringes filled with saline were occluded with a stopcock, and times to activation of the occlusion alarm were recorded. Statistical analysis was performed with Wilcoxon and Mann-Whitney U-tests, P < 0.05 was considered significant. RESULTS: The mean time to alarm activation was longer with lower infusion rates and larger syringes (P < 0.05). We recorded delays up to 117.3 +/- 9.4 min with 50 ml syringe at 0.5 ml x h(-1) and 15.0 +/- 7.1 min at 5 ml x h( 1). Syringe type had no effect on time to alarm activation (P > 0.05). The alarm on the JMS SP-500 pump was activated faster than the JMS SP-100 pump (P < 0.05). CONCLUSION: Our results showed that activation of occlusion alarms on both pumps takes a considerable time, and that the mean time to alarm activation was longer with low infusion rates and larger syringes. To reduce occlusion alarm delays smaller sized syringes with low compliance should be used and staff be alerted when using low flow rates with highly concentrated potent drugs. PMID- 15787920 TI - Umbilical vein catheterization under electrocardiogram guidance. AB - BACKGROUND: In the neonate, umbilical venous catheters (UVC) are inserted and advanced blindly to a predetermined length from the umbilicus. The reported rates for UVC misplacement into the liver (and occasionally the spleen) range from 20 to 37%. Radiographs are routinely used to confirm the positioning of UVCs. This involves movement of often critically ill infants, as well as radiation exposure. This pilot study examines the potential value of confirming UVC placement in neonates using ECG. METHODS: In critically ill neonates, a conductive Johans ECG adapter was connected to a UVC. A satisfactory tracing (lead II) was obtained (right arm lead connected to the adapter) when the UVC was filled with saline solution allowing the catheter tip to become a unipolar ECG electrode. The UVC was then advanced from the umbilicus until the tip reached the inferior vena cava (IVC) within the thoracic region, as demonstrated by appearance of normal sized QRS complexes with small P-waves. A small QRS indicated the catheter was below the diaphragm. The appearance of a tall positive P-wave indicated the tip was at the right atrium level. The UVC was then withdrawn until the P-wave size returned to normal. The final UVC position was later confirmed by X-ray. RESULTS: Eight neonates were studied. The figure shows typical ECG tracings when the UVC was placed in the liver, IVC, and right atrium, respectively. Three malpositioned catheters were detected (2 into liver and 1 into spleen). CONCLUSIONS: Based on these cases, the insertion of UVCs in neonates can be guided with ECG by observing sequential and characteristic alterations in P-waves and QRS complexes, thereby reducing the use of X-rays. In addition, this technique could prove to be beneficial in remote healthcare facilities where X-ray machines may not be readily available and quick intravenous access is required to transport sick neonates to major centers. PMID- 15787921 TI - A prospective study comparing the analgesic efficacy of levobupivacaine, ropivacaine and bupivacaine in pediatric patients undergoing caudal blockade. AB - BACKGROUND: The aim of our study was to compare postoperative analgesic efficacy, analgesic duration and motor blockade of levobupivacaine, ropivacaine and bupivacaine administered caudally in equal concentrations to children undergoing elective minor surgery. METHODS: In the study, 182 children, aged 1-7 years, undergoing either inguinal hernia repair or orchidopexy, were randomly allocated to one of the three groups. They received via a caudal extradural either 1 ml x kg(-1) levobupivacaine 0.2% (Group L) or 1 ml x kg(-1) ropivacaine 0.2% (Group R) or 1 ml x kg(-1) bupivacaine 0.2% (Group B). RESULTS: No statistically significant difference was noted in age, weight, duration of the operation or level of the caudal block between the groups. The onset of analgesia was significantly later after levobupivacaine. Postoperative pain scoring evaluated with Children's and Infant's Postoperative Pain Scale observational scale showed no statistical difference between groups. Median postoperative analgesia was 5.75 h (SEMed: +/- 0.65) in Group L, 5.7 h (SEMed: +/- 0.8) in Group R and 5.35 h (SEMed: +/- 1.3) in Group B the difference being statistically nonsignificant. CONCLUSIONS: The degree of motor block was significantly less after ropivacaine and levobupivacaine during the first 2 h postoperatively. PMID- 15787922 TI - Single-breath vital capacity rapid inhalation induction with sevoflurane: feasibility in children. AB - BACKGROUND: Single-breath vital capacity technique is currently administered for inhalation induction of anesthesia with sevoflurane in adults. Because sevoflurane is used in children, the aim of this open nonrandomized trial was to explore the feasibility and acceptance of this technique in midazolam premedicated patients aged from 4 to 15 years old. METHODS: A pediatric population (n = 118) was instructed in the vital capacity technique after their arrival in the induction room in a standardized and playful manner. Induction was performed with a circle-absorber breathing circuit, primed with sevoflurane 7% in 100% O2. Success of the single-breath vital capacity, delay of induction, hemodynamic and airway tolerance, acceptance by the children and side effects were analyzed. A multivariate logistic regression model was used to identify independent risk factors associated with the failure of the vital capacity technique. RESULTS: Single-breath vital capacity technique was achieved by 57% of the children. The success rate highly correlated with age and ranged from 10% in 4-5 years old to 75% at 11 years and 95% by 14 years. Other factors for success were cooperation and understanding. The need for more than two explanations of the technique was predictive of failure. Delays in the loss of the eyelash reflex and central pupil myosis were obtained in 34 s (18-50) and 242 s (145-278), respectively [median (interquartile ranges)]. Hemodynamic tolerance was good with few airway complications. CONCLUSION: Rapid anesthesia induction using a single breath technique with 7% sevoflurane is effective and well tolerated in children, particularly in those above 9 years of age, and in fact, success rate was markedly lower in the young age groups. PMID- 15787923 TI - Evaluation of satisfaction level by parents and children following pediatric anesthesia. AB - BACKGROUND: Research in the field of patient satisfaction first took place in the 1980s in the USA, and later in Europe, aimed at meeting competitive requirements in the field of health care. Very few studies of this type exist in regard to pediatric anesthesia. Our aim was to develop a rapidly interpretable questionnaire to measure the level of parental satisfaction when their children undergo surgery and provide information on those factors triggering anxiety in these children. METHODS: Over a period of 18 months 179 parents of children in pediatric surgery responded to 214 questionnaires designed to investigate principally the emotional/behavioral spheres as well as the comfort provided. RESULTS AND CONCLUSIONS: We found that parents generally expressed a high degree of satisfaction which was good and correlated significantly with environmental comfort and postoperative observations performed by anesthetists and nursing staff. In the children, lack of fear at the moment of being anesthetized, and lack of anxiety on the day preceding surgery, were attributed to the serenity transmitted by the anesthetist and nurses. Significant anxiety resulted from the fear of an unpleasant impact with the operating room. PMID- 15787924 TI - CT-guided lung biopsies in children: anesthesia management and complications. AB - We describe the anesthetic management of three children who underwent CT-guided lung biopsies and the complications associated with the procedure. We discuss the likely causes and recommend steps that would help decrease the risk of these complications during such a procedure. PMID- 15787925 TI - Ultrasound-guided central venous cannulation in a very small preterm neonate. AB - Percutaneous central venous cannulation of small infants is a challenging procedure. The use of ultrasound guidance has been shown to increase the success rate generally in children and to decrease the incidence of associated complications. To demonstrate that this technique is also suitable in very small infants we describe the case of a preterm neonate of 850 g body weight (BW), in which percutaneous central venous cannulation was performed successfully using ultrasound imaging for guidance. PMID- 15787926 TI - Argatroban for anticoagulation during cardiopulmonary bypass in an infant. AB - Heparin induced thrombocytopenia (HIT) is a rare, but potentially life threatening complication of heparin therapy. In patients with HIT, alternative means of anticoagulation are necessary. The authors present an infant with HIT who required anticoagulation during cardiopulmonary bypass for tricuspid valve excision in the treatment of bacterial endocarditis. The direct thrombin inhibitor, argatroban, was successfully used. Previous reports regarding the use of argatroban and other nonheparin anticoagulants for anticoagulation are reviewed and suggestions regarding argatroban dosing in infants are presented. PMID- 15787927 TI - Postoperative methemoglobinemia with associated G-6-P-D deficiency in infant cardiac surgery--enigmas in diagnosis and management. AB - Methemoglobinemia could cloud a turbulent postoperative course in infant cardiac surgery by interfering with the diagnosis of various critical conditions. Issues surrounding the treatment with methylene blue in the presence of glucose-6 phosphate dehydrogenase (G-6-P-D) deficiency are highlighted. PMID- 15787928 TI - Anesthesia for cleft lip surgery in a child with Seckel syndrome--a case report. AB - We describe the anesthetic management of a 5-year-old girl diagnosed with Seckel syndrome, for cleft lip surgery. The syndrome is characterized by severe intrauterine and postnatal growth retardation with proportional dwarfism, typical beak-like triangular nose, and mental retardation, accounting for its various synonyms such as bird-headed dwarfism, microcephalic primordial dwarfism, nanocephalic dwarfism and Seckel-type dwarfism. PMID- 15787929 TI - Management of the difficult pediatric airway with Shikani Optical Stylet. AB - The airways of children with many craniofacial and mandibulofacial malformations often present unique challenges to the anesthesiologists. We report the application of Shikani Optical Stylet (SOS) in facilitating the tracheal intubation of four children with history of difficult airway management. The SOS combines the benefits of the lightwand and fiberoptic bronchoscope. PMID- 15787930 TI - A case of supercarbia following pneumoperitoneum in an infant. AB - A 4-month-old healthy male infant underwent left herniotomy under general anesthesia with caudal block. Carbon dioxide (CO2) pneumoperitoneum was created through the left hernial sac for inspection of the right processus vaginalis. Episodes of desaturation associated with significant reduction in chest compliance were noted intraoperatively. This was overcome by increasing the inspired oxygen concentration (FiO2). The infant failed to regain consciousness and spontaneous respiration at the end of surgery. The chest compliance deteriorated further and clinically a CO2 pneumothorax (capnothorax) was suspected. The endtidal carbon dioxide (P(E)CO2) was initially low in the immediate postoperative period. Subsequent to the readministration of sevoflurane and manual ventilation with a Jackson Rees circuit, a sudden surge in P(E)CO2 with improvement of chest compliance was observed. At that time arterial blood gas (ABG) analysis revealed a PCO2 of 17.5 kPa (134 mmHg) and pH of 6.9. The causes of severe hypercarbia and the physiological changes observed in this infant are discussed. PMID- 15787931 TI - Anesthesia in a child with deletion 13q syndrome. PMID- 15787932 TI - Deep positioning of an esophageal temperature probe may lead to overestimation of core body temperature during laparoscopic Nissen fundoplication in infants. PMID- 15787934 TI - Difficult weaning: failure of conventional ventilation or inadequacy of PEEP. PMID- 15787935 TI - Classic Laryngeal Mask Airway in cardiac pediatric surgery. PMID- 15787936 TI - Alvimopan, a selective peripherally acting mu-opioid antagonist. AB - Alvimopan is a novel, peripherally acting mu-opioid antagonist that is being developed for the management of acute postoperative ileus and for the reversal of the delayed gastrointestinal and colonic transit that result in symptoms such as constipation, nausea and motility disorders in patients treated with opiate analgesics. There is a clinical need for effective medications for the treatment of postoperative ileus and opiate-induced constipation and other motility disorders. This review addresses the basic and applied pharmacology and current evidence for the use of the medication, alvimopan, in clinical gastroenterology. PMID- 15787937 TI - Altered neuroendocrine response and gastric dysmotility in the Flinders Sensitive Line rat. AB - In the search for animal models that can replicate some features of functional dyspepsia (FD) patients, we turned our interest to the Flinders Sensitive Line (FSL) rat. Gastric motility disturbances prevalent in FD patients as well as urine corticosterone and plasma prolactin were measured following buspirone challenge. Flinders Resistant Line (FRL) rat was used as control. The results show that the FSL rats have a disturbed gastric motility, reflected as both an increased gastric accommodation rate and gastric volume during gastric distension as well as a delayed gastric emptying, the latter possibly as a consequence of the former. Lipid administration resulted in a significant increase in maximal gastric volume only in the FRL rats. Both the corticosterone response to buspirone and the 24-h urinary output of corticosterone were normal in FSL rats. Similar to FD patients, the FSL rat showed supersensitivity to buspirone in the increase in prolactin release. Although FSL rats show some features similar to a subset of FD patients, the increased gastric accommodation contrasts to the reduced accommodation often seen in FD patients. Further studies are warranted to determine the relevance of this rat strain as a model for FD. PMID- 15787938 TI - Flow across the gastro-esophageal junction: lessons from the sleeve sensor on the nature of anti-reflux barrier. AB - The principles of fluid flow are critical to the understanding of flow across the gastro-esophageal (GE) junction or gastro-esophageal reflux. The mechanism by which sleeve sensor measures lower esophageal sphincter (LES) pressure is also based on the principle of fluid flow across the GE junction and can provide important insights into the nature of anti-reflux barrier. We present arguments regarding the role of the length of LES, intra-abdominal length of the LES, flap valve at the GE junction and the angle of His on the strength of anti-reflux barrier. Our conclusion is that in the presence of normal anatomy of the GE junction (absence of hiatus hernia) the pressure at the GE junction is the most important determinant of the strength of anti-reflux barrier. PMID- 15787939 TI - Mechanisms underlying duodeno-gastric reflux in man. AB - BACKGROUND: Recent studies suggest that duodeno-gastro-oesophageal reflux (DGER) contributes to the occurrence of reflux oesophagitis and Barrett's oesophagus. The mechanisms underlying duodeno-gastric reflux (DGR), a prerequisite for DGER, are poorly understood. AIMS: To study the occurrence of DGR in relation to interdigestive and postprandial gastroduodenal motility. SUBJECTS AND METHODS: Ten healthy subjects underwent stationary gastroduodenal manometry with simultaneous duodenal and antral Bilitec recording 4 h before and 5 h after ingestion of a liquid meal. Eight volunteers underwent the same study, with administration of erythromycin postprandially. RESULTS: During the interdigestive phase II, all volunteers had short DGR episodes. Postprandially, DGR occurred in all subjects, on average 39 +/- 28 min after the start of the meal, and was cleared from the stomach after 242 +/- 23 min. Induction of increased antral motility and of a premature phase III, by administration of erythromycin, was associated with faster gastric DGR clearance. However, there was no direct temporal relationship between erythromycin-induced gastric phase III and erythromycin-induced DGR clearance. CONCLUSION: In healthy subjects, duodenogastric reflux occurs sporadically in the interdigestive state and is a normal phenomenon in the postprandial period. Erythromycin induces faster clearance of DGR from the stomach, which depends on enhanced antral contractile activity rather than premature phase III. PMID- 15787940 TI - Measuring EGJ opening patterns using high resolution intraluminal impedance. AB - The aim of this study was to adapt impedance methodology to study esophagogastric junction (EGJ) sphincter opening and compare opening patterns of the EGJ during deglutitive LES relaxation (dLESR) and transient LES relaxation (tLESR). We studied eight healthy subjects with a novel 12-lumen combined impedance/manometry catheter, the main element of which was a 6 cm sleeve sensor with six side hole sensors and six impedance rings spaced at 1 cm increments along its length. Subjects underwent an air infusion protocol after standard assessment and data tracings and isocontour plots were analysed to assess opening characteristics of the EGJ during dLESRs and tLESRs. Our results revealed that during dLESR the opening pattern was top to bottom, occurred in 0-2.7 s and in 29 of 35 (83%) cases the leading edge of the bolus was liquid. Opening during tLESR began between -7.8 and +8.6 s relative to the onset of nadir LES relaxation. The opening pattern during tLESR was bottom to top, occurred in 0-7.7 s, and in 22 of 29 (76%) the leading edge was liquid. These results support that impedance monitoring can be adapted to identify sphincter opening, to distinguish sphincter opening from sphincter relaxation, and to determine luminal contents during the opening period. PMID- 15787941 TI - Unexpectedly low prevalence rates of IBS among adult Israeli Jews. AB - The irritable bowel syndrome (IBS) and other functional bowel disorders (FBDs) are common functional gastrointestinal disorders. The prevalence of IBS using Rome II criteria is generally lower than with previous criteria. The aim of this study was to determine the prevalence of IBS and other FBDs in the adult Israeli Jewish population, which has not been surveyed to date. A telephone survey was conducted on a representative sample of the study population provided by the Israel Ministry of the Interior. IBS and other FBDs were diagnosed by Rome II criteria. The study population was 981 individuals and the overall response rate was 54%. The mean age was 45.0 years and 55% were females. In all, 2.9% had IBS (females: 3.7%, males: 1.8%, P = 0.08). The rate increased to 4.1% when the Rome II diagnostic criteria were amended to include some chronic alternators who are not picked up by the original scoring system. Approximately 26% of the respondents had a functional lower gastrointestinal (GI) disorder (females: 32.1%, males: 17.7%, P < 0.0001). Prevalence rates for IBS among Israeli Jewish adults are lower than rates reported from most countries, despite the high level of stress resulting from Israel's geopolitical circumstances. Possible reasons for this low prevalence are discussed. PMID- 15787942 TI - The effects of the 5-HT3 antagonist, alosetron, on brain serotonin synthesis in patients with irritable bowel syndrome. AB - Serotonin (5-HT) plays an important role in the pathophysiology of irritable bowel syndrome (IBS). Using alpha-[(11)C]methyl-L-tryptophan-positron emission tomography (PET), it was demonstrated that brain 5-HT synthesis is increased in patients with IBS, in a gender-specific manner. The aims of the study were to evaluate the effects of alosetron on brain 5-HT synthesis in patients with IBS. Six male and five female non-constipation-predominant IBS patients were enrolled. The subjects received alosetron or a placebo for 14 days, separated by a 2-week washout period. On day 14, rectal distensions commenced just prior to the PET scan (which was performed for 80 min), and continued for 20-min periods. The functional images were analysed with SPM99. Alosetron vs placebo treatments, in a randomized, double-blinded, crossover manner, were studied. 5-HT synthesis was greater in several regions in the males than in the females during the alosetron treatment, whereas there was no region in which the females had greater synthesis. There were significant gender-treatment interactions of synthesis in the cingulate gyrus, caudate nucleus, globus pallidus, and cerebellum. The gender differences in the effect of alosetron on brain 5-HT synthesis may be related to the gender differences in the efficacy of alosetron. PMID- 15787943 TI - Effects of vasopressin and long pulse-low frequency gastric electrical stimulation on gastric emptying, gastric and intestinal myoelectrical activity and symptoms in dogs. AB - The aim of this study was to investigate the effect of vasopressin and long pulse low frequency gastric electrical stimulation (GES) on gastric emptying, gastric and intestinal myoelectrical activity and symptoms in dogs. The study was performed in eight healthy female dogs implanted with four pairs of gastric serosal electrodes and two pairs of small bowel serosal electrodes, and a duodenal fistula for the assessment of gastric emptying. Each dog was studied in three sessions on three separate days in a randomized order with recordings of gastric and small bowel slow waves. Each study session consisted of 30-min baseline, 30-min stimulation and 30-min recovery period. In sessions 1 and 2, infusion of either saline or vasopressin (0.75 U kg(-1) in 30 mL saline instilled in 30 min) was given during the second 30-min period. The protocol of session 3 was the same as session 2 except long pulse-low frequency GES was performed during the second 30-min period. It was found that: (i) Vasopressin significantly delayed gastric emptying 30 and 45 min after meal and GES did not improve the vasopressin induced delayed gastric emptying; (ii) Vasopressin induced gastric dysrhythmias and GES significantly improved vasopressin induced gastric dysrhythmia; (iii) Vasopressin also induced intestinal slow wave abnormalities but GES had no effect on vasopressin induced small bowel dysrhythmia; (iv) Vasopressin induced symptoms and behaviours suggestive of nausea that were not improved by GES. We conclude that: (i) Vasopressin delays gastric emptying and induces gastric and small bowel dysrhythmias and symptoms in the fed state, and (ii) long pulse-low frequency GES normalizes vasopressin induced gastric dysrhythmia with no improvement in gastric emptying or symptoms. PMID- 15787944 TI - Impaired gastric motor activity after abdominal surgery in rats. AB - Postoperative ileus (POI) is a transient bowel dysmotility that occurs following abdominal surgery. Several mechanisms have been proposed such as neural reflex and inflammatory changes. We focused on gastric motility after abdominal surgery in rats. To investigate the time course of gastric motility after surgery, gastric motility was continuously recorded before, during and after surgery. After laparotomy, terminal ileum was manipulated for 10 min. Gastric motility was recorded by a strain gauge transducer implanted on the serosal surface of the stomach. To investigate whether peripheral sympathetic nerve is involved in the pathogenesis of POI, effects of guanethidine and celiac ganglionectomy were tested on the postoperative gastric motility. Although isoflurane anaesthesia reduced the gastric motility to 40%, the motility recovered immediately when isoflurane was withdrawn. Intestinal manipulation reduced the postoperative gastric motility for 3-24 h after surgery, compared with preoperative levels. Guanethidine administration and celiac ganglionectomy restored the impaired gastric motility. Feeding increased the gastric motility in each group. It is suggested that the pathogenesis of postoperative gastric ileus induced by intestinal manipulation involves viscero-sympathetic pathways. Intestinal manipulation causes impaired gastric motility via inhibitory sympathetic efferent pathway. Feeding may improve the postoperative gastric motility. PMID- 15787945 TI - Role of oxidative stress in the pathogenesis of septic ileus in mice. AB - We investigated the role of oxidative stress in the pathogenesis of septic ileus. Sepsis was induced by intraperitoneal (i.p.) injection of lipopolysaccharides (LPS, 20 mg kg(-1)) in mice. The effect of two i.p. injections of superoxide dismutase [polyethylene glycol (PEG)-SOD, 4000 U kg(-1)] and catalase (PEG-CAT, 15,000 U kg(-1)) was investigated on gastric emptying, intestinal transit and total nitrite plasma concentrations. We also performed immunohistochemical experiments on gastric and ileal tissue. LPS significantly delayed gastric emptying and intestinal transit while plasma nitrite levels increased. Polyethylene glycol (PEG)-SOD reversed the endotoxin-induced delay in gastric emptying and improved the delay in intestinal transit without effect on plasma nitrite levels. PEG-CAT slightly improved the delay in gastric emptying without effect on intestinal transit. Immunohistochemistry showed the presence of nitrotyrosine (NT) and 4-hydroxy-2-nonenal (HNE) in the gastric and ileal mucosa of LPS-treated mice. Treatment with PEG-SOD or PEG-CAT of LPS mice diminished the presence of NT or HNE in both tissues. In addition, LPS induced a significant increase in inducible nitric oxide synthase (iNOS)-positive residential macrophages in the external musculature of stomach and ileum, which significantly decreased after PEG-SOD or PEG-CAT treatment. The present results support a role for oxidative and nitrosative stress in the pathogenesis of septic ileus in mice. PMID- 15787946 TI - A quantitative approach to recording peristaltic activity from segments of rat small intestine in vivo. AB - We have developed methods that allow correlation of propulsive reflexes of the intestine with measurements of intraluminal pressure, fluid movement and spatio temporal maps of intestinal wall movements for the first time in vivo. A segment of jejunum was cannulated and set up in a Trendelenburg recording system while remaining connected to the vascular and nerve supply of the anaesthetized rat. The resting intraluminal pressure in intact intestine was 2-4 mmHg. Hydrostatic pressures of 2, 4, 8 and 16 mmHg were imposed. At a baseline pressure of 4 mmHg, propulsive waves generated pressures of 9 +/- 1 mmHg, that progressed oral to anal at 2-5 mm s(-1). Individual propulsive waves propelled 0.8 +/- 0.4 mL of fluid. The frequency of propulsive waves increased with pressure, but peristaltic efficiency (mL per contraction) decreased with pressure increase between 4 and 16 mmHg. Atropine, as a bolus, transiently blocked peristalsis, but caused maintained block when infused. Hexamethonium blocked propulsive contractions. Inhibition of nitrergic transmission converted regular peristalsis to non propulsive contractions. These studies demonstrate the utility of an adapted Trendelenburg method for quantitative investigation of motility and pharmacology of enteric reflexes in vivo. PMID- 15787947 TI - Identification of galanin receptor 1 on excitatory motor neurons in the guinea pig ileum. AB - Exogenously administered galanin inhibits cholinergic transmission to the longitudinal muscle and reduces peristaltic efficiency in the guinea pig ileum with a mechanism partially mediated by galanin receptor 1 (GAL-R1). We investigated the effect of exogenous galanin 1-16, which has high affinity for GAL-R1, on the ascending excitatory reflex of the circular muscle elicited by radial distension in isolated segments of guinea pig ileum. We used a three compartment bath that allows dissecting the ascending pathway into the oral (site of excitatory motor neurons), intermediate (site of ascending interneurons) and caudal compartment (site of intrinsic primary afferent neurons). Galanin 1-16 (0.3-3 micromol L(-1)) applied to the oral compartment inhibited in a concentration-dependent manner the ascending excitatory reflex elicited by the wall distension in the caudal compartment. This effect was antagonized by the GAL R1 antagonist, RWJ-57408 (1 and 10 micromol L(-1)). By contrast, galanin 1-16 was ineffective when added to the intermediate or caudal compartment up to 3 micromol L(-1). GAL-R1 immunoreactive neurons did not contain neuron-specific nuclear protein, a marker for intrinsic primary afferent neurons. These findings indicate that GAL-R1s are present on motor neurons responsible for the ascending excitatory reflex, but not on ascending interneurons and intrinsic primary afferent neurons. PMID- 15787948 TI - Human mast cell mediator cocktail excites neurons in human and guinea-pig enteric nervous system. AB - Neuroimmune interactions are an integral part of gut physiology and involved in the pathogenesis of inflammatory and functional bowel disorders. Mast cells and their mediators are important conveyors in the communication from the innate enteric immune system to the enteric nervous system (ENS). However, it is not known whether a mediator cocktail released from activated human mast cells affects neural activity in the ENS. We used the Multi-Site Optical Recording Technique to image single cell activity in guinea-pig and human ENS after application of a mast cell mediator cocktail (MCMC) that was released from isolated human intestinal mucosa mast cells stimulated by IgE-receptor cross linking. Local application of MCMC onto individual ganglia evoked an excitatory response consisting of action potential discharge. This excitatory response occurred in 31%, 38% or 11% neurons of guinea-pig submucous plexus, human submucous plexus, or guinea-pig myenteric plexus, respectively. Compound action potentials from nerve fibres or fast excitatory synaptic inputs were not affected by MCMC. This study demonstrates immunoneural signalling in the human gut and revealed for the first time that an MCMC released from stimulated human intestinal mast cells induces excitatory actions in the human and guinea-pig ENS. PMID- 15787949 TI - Comparison between partial agonist (ME3412) and antagonist (alosetron) of 5 hydroxytryptamine 3 receptor on gastrointestinal function. AB - Therapeutic use of 5-hydroxytryptamine 3 (5-HT(3)) receptor antagonists for diarrhoea-predominant irritable bowel syndrome may be accompanied by constipation. We hypothesized that ME3412, 5-chloro-2-(1,4-diazacycloheptan-1-yl) 7-methylbenzoxazole, a novel partial agonist of the 5-HT(3) receptor, would minimize constipation without reducing antidiarrhoeal activity. Receptor binding studies showed that ME3412 is highly selective for the human 5-HT(3) receptor (K(i) = 1.51 nmol L(-1)). A 5-HT(3) receptor agonist, 2-methyl-5-HT, caused contractile response in the isolated guinea-pig ileum and accelerated secretion in the guinea-pig colonic mucosal preparation. ME3412 and 5-HT(3) receptor antagonist, alosetron, antagonized the 2-methyl-5-HT-induced responses with similar potency in insurmountable and surmountable manner, respectively. ME3412 caused weak agonism in isolated ileum strips and also in the colonic mucosa with intrinsic activity of 0.09 and 0.59, respectively. In conscious dogs, alosetron (3 microg kg(-1) i.v.) suppressed the migrating motor complex (MMC), whereas a relatively high dose (300 microg kg(-1)) of ME3412 was required for inhibition of MMC. ME3412 and alosetron suppressed 5-HT induced-diarrhoea in mice. In contrast, ME3412 did not significantly affect colonic propulsion compared with alosetron. These results imply that the partial agonist may relieve diarrhoea with low risk of inducing constipation. PMID- 15787953 TI - Making Medicaid a block grant program: an analysis of the implications of past proposals. AB - In 2003, President Bush proposed converting Medicaid from an entitlement to a block grant program. Similar ideas from President Reagan in 1981 and Congress in 1995 were introduced but not enacted. Block grants aim to provide greater federal budget certainty and a stronger state incentive to contain program costs. This paper compares the preestablished funding levels proposed in 1981 and 1995 with what actually happened to federal Medicaid spending. Its results show that previous block grant proposals' funding levels at the national and state levels were quite different from what was anticipated and what occurred. As a result, Medicaid probably could not--and cannot--maintain existing health coverage under a block grant financing structure. PMID- 15787950 TI - Contraction induced by glicentin on smooth muscle cells from the human colon is abolished by exendin (9-39). AB - Glicentin and glucagon-like peptide-1 (7-36) amide (GLP-1) are gut hormones released during digestion. Glicentin and GLP-1 slow down gastric emptying and glicentin can switch off the duodenojejunal fed motor pattern. The effect of glicentin on the motor activity of colon has never been reported in humans. Our aim was to determine if circular smooth muscle cells (SMC) from the human colon are target cells for glicentin or GLP-1, and if their motility is dependent upon these digestive hormones. METHODS: Twenty-two resections were performed on patients operated for colon adenocarcinoma. The SMC were isolated from colonic circular muscle layer and cell contraction was assessed. RESULTS: Glicentin caused a dose-related contraction of SMC, when GLP-1 determined a contraction of weak amplitude. Exendin-(9-39), described as a GLP-1 receptor antagonist, inhibited contraction due to glicentin or GLP-1. In contrast, on antral SMC from rabbit, GLP-1 exerts neither relaxation nor contraction; however, exendin-(9-39) dose dependently reduced the contractile activity of glicentin [glicentin EC(50) = 5 pM, exendin-(9-39) pA(2) = -9.36]. CONCLUSIONS: The circular muscle from the human colon is a target tissue for glicentin and GLP-1. Whereas glicentin is a long-life digestive hormone which would contribute to segmental contraction, the biological activity of GLP-1 remains unknown on this tissue. On the digestive smooth muscle, exendin-(9-39) behaved as an antagonist for two members of the glucagon-receptor family, GLP-1 and glicentin. PMID- 15787952 TI - Mortality of white Americans, African Americans, and Canadians: the causes and consequences for health of welfare state institutions and policies. AB - The life expectancy of African Americans has been substantially lower than that of white Americans for as long as records are available. The life expectancy of all Americans has been lower than that of all Canadians since the beginning of the 20th century. Until the 1970s this disparity was the result of the low life expectancy of African Americans. Since then, the life expectancy of white Americans has not improved as much as that of all Canadians. This article discusses two issues: racial disparities in the United States, and the difference in life expectancy between all Canadians and white Americans. Each country's political culture and institutions have shaped these differences, especially national health insurance in Canada and its absence in the United States. The American welfare state has contributed to and explains these differences. PMID- 15787954 TI - Welfare reform and substance abuse. AB - The 1996 Personal Responsibility and Work Opportunity Reconciliation Act (PRWORA) changed the nature, purpose, and financing of public aid. Researchers, administrators, and policymakers expressed special concern about the act's impact on low-income mothers with substance use disorders. Before PRWORA's passage, however, little was known about the true prevalence of these disorders among welfare recipients or about the likely effectiveness of substance abuse treatment interventions for welfare recipients. Subsequent research documented that substance abuse disorders are less widespread among welfare recipients than was originally thought and are less common than other serious barriers to self sufficiency. This research also showed significant administrative barriers to the screening, assessment, and referral of drug-dependent welfare recipients. This article summarizes current research findings and examines implications for welfare reform reauthorization. PMID- 15787956 TI - Drug insurance utilization management policies and "reference pricing": an illustrated commentary on the article by Vittorio Maio and colleagues. PMID- 15787955 TI - Pharmacy utilization and the Medicare Modernization Act. AB - To control expenditures and use medications appropriately, the Medicare drug coverage program has established pharmacy utilization management (PUM) measures. This article assesses the effects of these strategies on the care of seniors. The literature suggests that although caps on drug benefits lower pharmaceutical costs, they may also increase the use of other health care services and hurt health outcomes. Our review raises concerns regarding the potential unintended effects of the Medicare drug program's PUM policies for beneficiaries. Therefore, the economic and clinical impact of PUM measures on seniors should be studied further to help policymakers design better drug benefit plans. PMID- 15787958 TI - Comment on "Diffusion of innovations in service organizations: systematic review and recommendations". PMID- 15787957 TI - Exporting the Buyers Health Care Action Group purchasing model: lessons from other communities. AB - When first implemented in Minneapolis and St. Paul, Minnesota, the Buyers Health Care Action Group's (BHCAG) purchasing approach received considerable attention as an employer-managed, consumer-driven health care model embodying many of the principles of managed competition. First BHCAG and, later, a for-profit management company attempted to export this model to other communities. Their efforts were met with resistance from local hospitals and, in many cases, apathy by employers who were expected to be supportive. This experience underscores several difficulties that appear to be inherent in implementing purchasing models based on competing care systems. It also, once again, suggests caution in drawing lessons from community-level experiments in purchasing health care. PMID- 15787961 TI - De novo design of a pentameric coiled-coil: decoding the motif for tetramer versus pentamer formation in water-soluble phospholamban. AB - Water-soluble phospholamban (WSPLB) is a designed, water-soluble analogue of the pentameric membrane protein phospholamban (PLB), which contains the same core and interhelical residues as PLB, with only the solvent-exposed positions mutated. WSPLB contains the same secondary and quaternary structure as PLB. The hydrophobic cores of PLB and WSPLB contain Leu and Ile at the a- and d-positions of a heptad repeat (abcdefg) from residues 31-52, while residues 21-30 are rich in polar amino acids at these positions. While the full-length WSPLB forms pentamers in solution, truncated peptides lacking residues 21-30 are largely tetrameric. Thus, truncation of residues 1-20 promotes a switch from pentamer to tetramer formation. Here, the motifs for WSPLB pentamerization were elucidated by characterizing a series of peptides, which were progressively truncated in this polar 'switch' region. When fully present, the 'switch' region promotes pentamer formation in WSPLB, by destabilizing a more stable tetrameric species which exists in its absence. We find that the burial of hydrogen bonding residues from 21 to 30 drives WSPLB from a tetramer to a pentamer, with direct implications for coiled-coil design. PMID- 15787962 TI - Structure-activity relationships of arodyn, a novel acetylated kappa opioid receptor antagonist. AB - We previously reported that the novel dynorphin A (Dyn A, Tyr-Gly-Gly-Phe-Leu-Arg Arg-Ile-Arg-Pro-Lys-Leu-Lys-Trp-Asp-Asn-Gln) analog arodyn (Ac[Phe(1,2,3),Arg(4),d-Ala(8)]Dyn A-(1-11)NH(2), Bennett, M.A., Murray, T.F. & Aldrich, J.V. (2002) J. Med. Chem. vol. 45, pp. 5617-5619) is a kappa opioid receptor-selective peptide [K(i)(kappa) = 10 nm, K(i) ratio (kappa/mu/delta) = 1/174/583] which exhibits antagonist activity at kappa opioid receptors. In this study, a series of arodyn analogs was prepared and evaluated to explore the structure-activity relationships (SAR) of this peptide; this included an alanine scan of the entire arodyn sequence, sequential isomeric d-amino acid substitution in the N-terminal 'message' sequence, NMePhe substitution individually in positions 1-3, and modifications in position 1. The results for the Ala substituted derivatives indicated that Arg(6) and Arg(7) are the most important residues for arodyn's nanomolar binding affinity for kappa opioid receptors. Ala substitution of the other basic residues (Arg(4), Arg(9) and Lys(11)) resulted in lower decreases in affinity for kappa opioid receptors (three- to fivefold compared with arodyn). Of particular interest, while [Ala(10)]arodyn exhibits similar kappa opioid receptor binding as arodyn, it displays higher kappa vs. mu opioid receptor selectivity [K(i) ratio (kappa/mu) = 1/350] than arodyn because of a twofold loss in affinity at mu opioid receptors. Surprisingly, the Tyr(1) analog exhibits a sevenfold decrease in kappa opioid receptor affinity, indicating that arodyn displays significantly different SAR than Dyn A; [Tyr(1)]arodyn also unexpectedly exhibits inverse agonist activity in the adenylyl cyclase assay using Chinese hamster ovary cells stably expressing kappa opioid receptors. Substitution of NMePhe in position 1 gave [NMePhe(1)]arodyn which exhibits high affinity [K(i)(kappa) = 4.56 nm] and exceptional selectivity for kappa opioid receptors [K(i) ratio (kappa/mu/delta) = 1/1100/>2170]. This peptide exhibits antagonistic activity in the adenylyl cyclase assay, reversing the agonism of 10 nm Dyn A-(1-13)NH(2). Thus [NMePhe(1)]arodyn is a highly kappa opioid receptor-selective antagonist that could be a useful pharmacological tool to study kappa opioid receptor-mediated activities. PMID- 15787963 TI - Roles of residues 3 and 4 in cyclic tetrapeptide ligand recognition by the kappa opioid receptor. AB - A series of cyclic, disulfide- or dithioether-containing tetrapeptides based on previously reported potent mu- and delta-selective analogs has been explored with the aim of improving their poor affinity to the kappa-opioid receptor. Specifically targeted were modifications of tetrapeptide residues 3 and 4, as they presumably interact with residues from transmembrane helices 6 and 7 and extracellular loop 3 that differ among the three receptors. Accordingly, tetrapeptides were synthesized with Phe(3) replaced by aliphatic (Gly, Ala, Aib, Cha), basic (Lys, Arg, homo-Arg), or aromatic sides chains (Trp, Tyr, p NH(2)Phe), and with d-Pen(4) replaced by d-Cys(4), and binding affinities to stably expressed mu-, delta-, and kappa-receptors were determined. In general, the resulting analogs failed to exhibit appreciable affinity for the kappa receptor, with the exception of the tetrapeptide Tyr-c[d-Cys-Phe-d-Cys]-NH(2), cyclized via a disulfide bond, which demonstrated high binding affinity toward all opioid receptors (Ki(mu) = 1.26 nm, Ki(delta) = 16.1 nm, Ki(kappa) = 38.7 nm). Modeling of the kappa-receptor/ligand complex in the active state reveals that the receptor-binding pocket for residues 3 and 4 of the tetrapeptide ligands is smaller than that in the mu-receptor and requires, for optimal fit, that the tripeptide cycle of the ligand assume a higher energy conformation. The magnitude of this energy penalty depends on the nature of the fourth residue of the peptide (d-Pen or d-Cys) and correlates well with the observed kappa-receptor binding affinity. PMID- 15787965 TI - Exploring the stereochemical requirements for protease inhibition by ureidopeptides. AB - A novel 'ureidopeptide' substrate analog inhibitor of the HIV-1 protease, created by substitution of a urea for the scissile amide bond of a hexapeptide substrate, was synthesized and tested for inhibition of HIV-1 protease. This inhibitor was designed as a stereochemical mutant of an earlier ureidopeptide inhibitor in which the P1' phenylalanine residue was changed from an l-isomer to a d-isomer. This was done in an attempt to increase binding to the enzyme by compensating for a lengthening of the peptide backbone. The inhibitor was synthesized from two protected tripeptide precursors using an oxidative Hoffmann rearrangement of a C terminal peptide amide. The new inhibitor was found to inhibit HIV-1 protease with an observed IC(50) of 47 mum. PMID- 15787964 TI - Alginate as an auxiliary bacterial membrane: binding of membrane-active peptides by polysaccharides. AB - The chronicity of Pseudomonas aeruginosa infections in cystic fibrosis (CF) patients is characterized by overproduction of the exopolysaccharide alginate, in which biofilm bacteria are embedded. Alginate apparently contributes to the antibiotic resistance of bacteria in this form by acting as a diffusion barrier to positively charged antimicrobial agents. We have been investigating cationic antimicrobial peptides (CAPs) (prototypic sequence: KKAAAXAAAAAXAAWAAXAAAKKKK NH(2), where X is any of the 20 commonly occurring amino acids) that were originally designed as transmembrane mimetic peptides. Peptides of this group above a specific hydrophobicity threshold insert spontaneously into membranes and have antibacterial activity at micromolar concentrations. While investigating the molecular basis of biofilm resistance to peptides, we found that the anionic alginate polysaccharide induces conformational changes in the most hydrophobic of these peptides typically associated with insertion of such peptides into membrane environments [Chan et al., J. Biol. Chem. (2004) vol. 279, pp. 38749-38754]. Through a combination of experiments measuring release of the fluorescent dye calcein from phospholipid vesicles, peptide interactions with vesicles in the presence and absence of alginate, and affinity of peptides for alginate as a function of net peptide core hydrophobicity, we show here that alginate offers a microenvironment that provides a protective mechanism for the encased bacteria by both binding and promoting the self-association of the CAPs. The overall results indicate that hydrophilic alginate polymers contain a significant hydrophobic compartment, and behave as an 'auxiliary membrane' for bacteria, thus identifying a unique protective role for biofilm exopolysaccharide matrices. PMID- 15787966 TI - Liposomal cargo unloading induced by pH-sensitive peptides. AB - Amphiphilic peptides designed to have a pH-dependent conformational change and membrane activity are described. At physiologic pH, the peptides would exist in a random coil conformation, but at endosomal pH values they would switch to amphiphilic alpha-helices, disrupt membranes, and release liposomal contents. A series of peptides have been investigated that contain a high percentage of Glu residues for the pH-induced conformational switch, and Leu residues for optimal lipid binding. Circular dichroism (CD) results in aqueous and liposomal environments were performed and demonstrate a pH-dependent shift to helicity upon acidification. Liposomal release data at neutral and acidic pH, also document the success of this design strategy. PMID- 15787967 TI - Water-soluble prodrugs of cyclosporine A with tailored conversion rates. AB - A novel type of water-soluble prodrugs of cyclosporine A (CsA) is described, featuring a modular system of an enzyme-cleavable group, a solubilizing moiety and a chemodegradable spacer attached to the hydroxyl function of (4R)-4-[(E)-2 butenyl]-4-,N-dimethyl-l-threonine (MeBmt)-1 of CsA. The chemical synthesis of these double prodrugs proceeds in high yield and purity and allows for a systematic study of the influence of the structural parameters upon physicochemical and pharmacological properties. The evaluation of the chemical and enzymatic stability results in differential values of the conversion rates (minutes to several hours) in support of an enzyme-triggered release of the parent drug as the rate-limiting step. In vitro studies show that the designed prodrug systems can be regarded as soft prodrugs in being devoid of cyclophiline A (CypA) binding and that complete conversion to the parent drug occurs in whole rat blood, setting the stage for therapeutic use. PMID- 15787968 TI - Immunostimulants and Toll-like receptor ligands obtained by screening combinatorial lipopeptide collections. AB - Synthetic lipopeptides carrying the head group of bacterial lipoproteins are specific ligands of Toll-like receptors (TLR). The three fatty acids containing lipopeptides with the tripalmitoyl-S-glyceryl-cysteinyl N-terminus (Pam(3)Cys) are agonists of TLR2. The structurally related lipopeptides with a head group lacking the fatty acyl residue at the amino-terminus (Pam(2)Cys) stimulate TLR2 and 6. To investigate the influence of the peptide chain of lipohexapeptides with a free N-terminus with regard to their ability to enhance B-cell proliferation, a randomized S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-(R)-cysteinyl-pentapeptide amide collection Pam(2)CysXXXXX and 5 x 19 subcollections (Pam(2)CysOXXXX, Pam(2)CysXOXXX, Pam(2)CysXXOXX, Pam(2)CysXXXOX, Pam(2)CysXXXXO, O: all protein amino acids except Cys) were prepared by parallel solid-phase synthesis. The collection represents synthetic lipopeptide analogues of the numerous bacterial lipoproteins and of mycoplasma lipoprotein. Each of the 95 subcollections is characterized by one defined and four degenerated amino acid positions thus comprising 19(4) individual lipopeptides with free N-terminal amino groups. High performance liquid chromatography electrospray mass spectrometry (HPLC-ESI-MS) was applied for the analytical characterization of the lipohexapeptide amide subcollections and for the individual lipohexapeptide amides. The subcollections were tested for polyclonal activation of murine spleen cells, deconvolution led to highly active single S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-(R)-cysteinyl pentapeptide amides. PMID- 15787969 TI - Use of the excluded protecting group (EPG) method for peptide synthesis. AB - The excluded protecting group (EPG) method has been used for the solution synthesis of several peptides including Merrifield's Model Tetrapeptide, linear antamanide and an analogue of magainin-1, [Ala(19), Asn(22)]magainin-1. In the approach reported, the C-terminal amino acid is esterified to the 2-position of cholestane as the [2s,3s]iodohydrin ester and the penultimate amino acid added to the aminoacyl-steroid as the Fmoc-pentafluorophenyl-ester. The Fmoc group is removed with Et(2)NH/DMF ( approximately 15% v/v) and, after evaporation to approximately 10 mL, the solution chromatographed on Sephadex LH-20 in DMF. The dipeptidyl-steroid elutes as the free amine well separated from other reaction mixture components. Fractions containing the dipeptide, as determined by counting and TLC, are pooled and reacted with the next Fmoc-amino acid-pentafluorophenyl ester in the sequence. Repetition of the deprotection/purification/reaction cycle yields the fully protected peptide. On completion of the synthesis, the cholestane iodohydrin ester is selectively removed by treatment with Zn degrees /AcOH to yield the peptide with intact alpha-amino and side chain protecting groups. Global deprotection is achieved with HF. All intermediates from the syntheses reported were characterized. The magainin analogue was shown to have full biologic activity. The Fmoc iodohydrin esters of 16 of the 20 proteogenic amino acids have been prepared and characterized for use as the C-terminal amino acids in other EPG syntheses. PMID- 15787970 TI - Preparation of protected peptidyl thioester intermediates for native chemical ligation by Nalpha-9-fluorenylmethoxycarbonyl (Fmoc) chemistry: considerations of side-chain and backbone anchoring strategies, and compatible protection for N terminal cysteine. AB - Native chemical ligation has proven to be a powerful method for the synthesis of small proteins and the semisynthesis of larger ones. The essential synthetic intermediates, which are C-terminal peptide thioesters, cannot survive the repetitive piperidine deprotection steps of N(alpha)-9-fluorenylmethoxycarbonyl (Fmoc) chemistry. Therefore, peptide scientists who prefer to not use N(alpha)-t butyloxycarbonyl (Boc) chemistry need to adopt more esoteric strategies and tactics in order to integrate ligation approaches with Fmoc chemistry. In the present work, side-chain and backbone anchoring strategies have been used to prepare the required suitably (partially) protected and/or activated peptide intermediates spanning the length of bovine pancreatic trypsin inhibitor (BPTI). Three separate strategies for managing the critical N-terminal cysteine residue have been developed: (i) incorporation of N(alpha)-9-fluorenylmethoxycarbonyl-S (N-methyl-N-phenylcarbamoyl)sulfenylcysteine [Fmoc-Cys(Snm)-OH], allowing creation of an otherwise fully protected resin-bound intermediate with N-terminal free Cys; (ii) incorporation of N(alpha)-9-fluorenylmethoxycarbonyl-S triphenylmethylcysteine [Fmoc-Cys(Trt)-OH], generating a stable Fmoc-Cys(H) peptide upon acidolytic cleavage; and (iii) incorporation of N(alpha)-t butyloxycarbonyl-S-fluorenylmethylcysteine [Boc-Cys(Fm)-OH], generating a stable H-Cys(Fm)-peptide upon cleavage. In separate stages of these strategies, thioesters are established at the C-termini by selective deprotection and coupling steps carried out while peptides remain bound to the supports. Pilot native chemical ligations were pursued directly on-resin, as well as in solution after cleavage/purification. PMID- 15787971 TI - Characterization of interactions of Na+/H+ exchanger regulatory factor-1 with the parathyroid hormone receptor and phospholipase C. AB - The Na(+)/H(+) exchanger regulatory factor-1 (NHERF1) is a molecular scaffold important for the signaling of the G-protein coupled receptor for the parathyroid hormone (PTH1R). The two PDZ (PSD-95, Discs-large, ZO1) domains of NHERF1 through association with the C-termini of PTH1R and phospholipase C enhance the signaling pathway associated with PTH. To examine these interactions, we have produced the individual PDZ1 and PDZ2 domains as well as the tandem PDZ1-PDZ2 domains (PDZ12) of NHERF1 and have characterized the binding affinities of the C-terminal motifs of PTH1R and PLCbeta using fluorescence anisotropy. Circular dichroism indicates that the PDZ1 and PDZ2 are properly folded. Based on fluorescence anisotropy we find that the C-terminus of PTH1R, containing ETVM, has similar affinities (approximately 10 microm) for both PDZ1 and PDZ2. The PTH1R displayed reduced binding affinity for the tandem PDZ12 (16 microm) compared with the individual domains or a solution of equal molar concentrations of PDZ1 and PDZ2 (5.8 microm), suggesting negative cooperativity between the PDZ domains or intervening region. The C-termini of PLCbeta (both beta1 and beta2 isozymes were examined, containing DTPL and ESRL, respectively) displayed a diminished affinity for PDZ2 (approximately 30 microm) over that of PDZ1 (approximately 8 microm). Finally, we demonstrate trans PDZ1-PDZ2 association that is enhanced in the presence of the C terminus of PTH1R or PLCbeta, suggesting oligomerization of NHERF as a mode for enhancing the signaling associated with PTH. PMID- 15787972 TI - Analysis of ligand-receptor cross-linked fragments by mass spectrometry. AB - G-protein coupled receptors (GPCRs) are a class of integral membrane receptor proteins that are characterized by a signature seven-transmembrane (7-TM) configuration. The alpha-factor receptor (Ste2p) from Saccharomyces cerevisiae is a GPCR that, upon binding of a peptide ligand, transduces a signal to initiate a cascade of events leading to the mating of haploid yeast cells. This study summarizes the application of affinity purification and of matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) experiments using biotinylated photoactivatable alpha-factor analogs. Affinity purification and enrichment of biotinylated peptides by monomeric avidin beads resulted in mass spectrometric detection of specific signals corresponding to cross-linked fragments of Ste2p. Data obtained from cyanogen bromide (CNBr) fragments of receptor cross-linked to an alpha-factor analog with the photoaffinity group p-benzoyl-l-phenylalanine on position 1 were in agreement with the previous results reported by our laboratory suggesting the cross-linking between position 1 of alpha-factor and a region of Ste2p covering residues 251-294. PMID- 15787973 TI - Dioxin residues in the edible tissue of finishing pigs after dioxin feeding. AB - The aim of this study was to determine the contamination of finishing pigs with polychlorinated dibenzodioxins and dibenzofurans (PCDD/Fs) after feeding either uncontaminated feed or feed contaminated with 0.75, 2 or 4 ng/kg toxic equivalents (TEQ; calculated by multiplying individual congener concentrations by congener-specific toxicity equivalency factors). The feed was mixed with pure substances of PCDD/Fs to get the intended contamination. Five groups of six piglets each were fed contaminated feed, one group of five piglets served as control. One group was fed contaminated feed (4 ng TEQ/kg) only for the rearing period (6 weeks), and another group for the first 8 weeks of the fattening period (4 ng TEQ/kg feed). The other groups received the contaminated feed during the 12 weeks fattening period. After slaughtering, the edible parts of the belly, loin and fore-end were collected and homogenized. The samples of group 2 and 4a were investigated uncooked as well as roasted. Fattening yield and feed conversion (kg feed/kg weight gain) of the animals of all groups were in the normal range (final weight 109.7 kg; feed conversion 2.55-2.69 kg). The PCDD/F-content in 1 kg fat of the belly, loin and fore-end in relation to the intake was between 0.016 (4 ng TEQ/kg feed for a 6-weeks rearing period) and 1.39% (fore-end; 2 ng TEQ/kg feed for 12-weeks fattening period). There was a decrease in dioxin residues after a 12-weeks period but not after a 4-weeks period of feeding an uncontaminated feed. When feed contaminated with 0.75, 2 and 4 ng TEQ/kg was given for a 12-weeks fattening period, the residue concentrations of PCDD/F-TEQ in 1 kg belly was 0.455, 1.07 and 1.55 ng, in 1 kg fore-end 0.04 ng, 0.32 ng and 0.34 ng and in 1 kg loin 0.015 ng, 0.07 ng and 0.30 ng respectively. Roasting had no influence on the dioxin-residues. The residues per g belly fat exceed the maximum limits for dioxin in food of 0.6 pg WHO-PCDD/F-TEQ/g fat (EC Recommendations 2002/201/EC), when feed containing 0.75 ng PCDD/F-TEQ/kg is given for 12 weeks. When feed containing 0.4 ng TEQ (maximum content; EC recommendations 2002/201/EC) is given for 12 weeks, approximately 0.55 pg TEQ/g fat can be expected in the food. This value is within the action level of 0.6 pg/g fat of porks. In conclusion, the results of the study allow prediction of dioxin residues in the edible tissue of pork, if the feed contamination is known and the amount of feed intake can be estimated. PMID- 15787974 TI - Dioxin concentration in milk and tissues of cows and sheep related to feed and soil contamination. AB - If cows or sheep are grazing on areas known for a higher dioxin level of the soil, respectively, dry cows are fed with feedstuffs produced on such areas, the dioxin concentration in food, in this case in cows' milk and livers of sheep could reach or exceed maximum levels. Furthermore large amounts of polychlorinated dibenzo-p-dioxins/polychlorinated dibenzo-p-furans (PCDD/F) can be excreted with the faeces. But there are chances for farmers to prevent or at least to minimize the dioxin load of feed and with it of food (including the harvesting technique and the feeding strategy). PMID- 15787975 TI - Carry over (transfer) of feed-borne acrylamide into eggs, muscle, serum, and faeces - a pilot study with Japanese quails (Coturnix coturnix japonica). AB - Two groups of 10 quails each were fed for 30 days on a diet containing either 663 microg feed-borne acrylamide (AA)/kg feed (group B) or 2472 microg AA/kg feed (group C) respectively. The concentrations of AA present in excreta and eggs were compared with those measured in eggs and excreta obtained from a control group (group A) fed on a diet low in AA (<10 microg/kg). The results clearly showed that the eggs of quails in group C contained approximately 53-112 microg AA/kg based on dry weight, while eggs collected from group A did not contain AA (limit of detection in eggs 3.5 microg/kg). Nearly 5% of the AA fed was recovered in the excreta. The AA concentrations in liver, muscle and serum were detectable, but below the limit of quantitation. PMID- 15787976 TI - Investigations on prophylactic effects of coarse feed structure and/or potassium diformate on the microflora in the digestive tract of weaned piglets experimentally infected with Salmonella Derby. AB - The purpose of this study was to examine potential prophylactic effects of a coarse feed structure and/or potassium diformate (KDF as feed additive) addition to diets on the microflora of the digestive tract in reared piglets experimentally infected with Salmonella Derby. The results show that coarse grinding as well as KDF addition are able to influence positively the intestinal flora and are capable to reduce Salmonella excretion of infected piglets. Coarse grinding of main ingredients (e.g. cereals) led to an increased number of lactobacilli as well as to higher counts of Gram-positive coccoid bacteria in the colon chyme, while KDF resulted in a tendency towards lower counts of Escherichia coli within the digestive tract. Moreover, a combination of both treatments influenced the composition of the gastrointestinal flora quite positively. Furthermore, the combination of these dietetic measures resulted in a reduced Salmonella excretion rate, shorter Salmonella shedding period and a reduced translocation of Salmonella within the infected piglets. The positive effects of combining both treatments led to a significantly reduced spreading of Salmonella within the group of pigs, which might be used to diminish Salmonella prevalence in pig production. PMID- 15787977 TI - Effect of combinations of organic acids in weaned pig diets on microbial species of digestive tract contents and their response on digestibility. AB - The effect of combinations of organic acids on digestive microbial population and total tract digestibility of piglets was studied. Thirty 19-21 days weaned pigs (5.4 +/- 0.23 kg, six pigs per treatment) were given a mixed feed with 10 ml water/kg (CTL) or 200 mEq/kg of formic acid (FOR), 1:1 formic:fumaric (FOFU), 1:1 formic:lactic (FOLA) or 2:1 formic:lactic (2FOLA). After 6-8 days, animals were slaughtered. In gastric contents, pH was higher (p = 0.01) with FOFU and lactobacilli tended (p = 0.08) to be lower with FOFU than CTL and FOLA, but coliform counts did not differ (p = 0.14). Acetate proportion was lower (p = 0.03), and propionate (p = 0.05) and butyrate (p = 0.01) higher, with FOFU than CTL, FOLA and 2FOLA. Intestinal coliform counts were higher (p = 0.03) with CTL than FOLA and 2FOLA, but there were no differences on lactobacilli. In intestinal contents, acetate tended (p = 0.06) to be lower with FOR than FOLA and 2FOLA, and butyrate was higher (p = 0.001) with FOR. Although not significantly, dry matter digestibility was 0.03-0.05 lower with CTL than with the other treatments. Combinations of organic acids in piglet diets modify gastric and intestinal microflora, the mixtures of formic:lactic appearing as the most interesting. PMID- 15787978 TI - Effects of a supranutritional dose of selenate compared with selenite on insulin sensitivity in type II diabetic dbdb mice. AB - The present study was performed to examine the mechanism by which selenate ameliorates the insulin sensitivity in type II diabetic dbdb mice. Therefore, 21 adult female dbdb mice were randomly assigned to three experimental groups (0Se, SeIV and SeVI) with seven animals per group. Mice of group 0Se were fed with a selenium-deficient diet (<0.02 mg Se/kg) based on wheat and torula yeast for 8 weeks whereas the mice of groups SeIV (selenite) and SeVI (selenate) were fed with sodium selenite and sodium selenate (up to 35% of the LD(50) for mice in eighth week), in addition to the diet by daily tube feeding. Eight weeks of selenate application led to significantly elevated insulin sensitivity in comparison with selenium deficiency and selenite application. The activity of cytosolic protein tyrosine phosphatases (PTPs) as important negative regulators of insulin signalling was reduced from 53.8% to 22.5% in the liver and skeletal muscle of selenate-treated mice in comparison with the selenium deficient and selenite-treated controls, suggesting an inhibition of PTPs by intermediary selenate metabolites. In an additional in vitro inhibition study, selenate (oxidation state +VI) did not inhibit PTP activity. Selenium metabolites in the oxidation state +IV were found to be the actual inhibitors of PTP activity. In conclusion, the results of the present study show that one possible mechanism by which supranutritional selenate doses enhance insulin sensitivity in type II diabetic dbdb mice is based on the inhibition of PTPS as negative regulators of insulin signalling. Moreover the cellular metabolism of selenate including its intermediary reduction to the oxidation state +IV seems to play a crucial role during this process. PMID- 15787979 TI - Experiments to optimize enzyme substitution therapy in pancreatic duct-ligated pigs. AB - Ligation of the pancreatic duct in pigs leads to severe maldigestion and malabsorption of crude nutrients. Supplementation with 24 capsules of Creon (Solvay Pharmaceuticals GmbH, Hannover, Germany) per meal led to an increased digestibility of crude nutrients. With regard to optimization of the treatment of EPI no essential improvements can be achieved by adding omeprazol or lecithin to the diet. In pancreatic duct-ligated pigs the isolated addition of omeprazol led to an increase of the pre-caecal digestibility of crude fat and organic matter. With additional enzyme substitution, the application of omeprazol did not result in an improved fat digestibility. Isolated addition of lecithin to the diet resulted in a reduced total digestibility of crude fat. Offering the diet twice a day and using a higher frequency of enzyme applications (four or six instead of only two applications) had no effects on the digestibilty of crude fat or organic matter. According to the observations in pancreatic duct-ligated pigs, the addition of missing enzymes to the diet led to the best treatment results in EPI. Administration of omeprazol or a higher feeding frequency as well as the application of enzymes in small proportion of the whole meal or dosages given consecutively over the day showed no advantages. Furthermore, the present study suggests that the addition of lecithin cannot be recommended in EPI, when given diets with butter as the predominant fat source as in human dietetics. PMID- 15787980 TI - Digestibility rates of major and trace elements in pancreatic duct-ligated pigs. AB - Ligation of pancreatic duct in pigs leads to a severe maldigestion of calcium and magnesium. Substitution of missing enzymes results in a 'normalization' of the digestibility rates of these elements. In comparison to controls the pre-caecal phosphorus digestibility decreased, but the total digestibility rates increased, after ligation of the pancreatic duct. Furthermore, the total amount of absorbed phosphorus was significantly higher in pancreatic duct ligated pigs. Further studies have to be carried out to investigate, whether phosphorus in exocrine pancreatic insufficiency is excreted with urine, as in this study the observed Ca:P-ratio in blood was in a physiological range. Without a forced renal excretion of phosphorus, consequences and risks (e.g. secondary hyperparathyroidism) of the regulation have to be considered. Regarding the elements sodium, potassium and chloride, an increased faecal excretion could be observed in pancreatic duct ligated pigs. As a substitution with enzyme products led to digestibility rates similar to those in controls, no losses of electrolytes have to be feared. Even though pancreatic juice seems to have influences on the digestibility of investigated trace elements (copper, zinc, iron and manganese), it did not lead to severe imbalances in the corresponding mineral metabolism. PMID- 15787981 TI - Influence of different calcium concentrations in the diet on bone metabolism in growing dairy goats and sheep. AB - The purpose of this study was to investigate, if different Ca concentrations in diets have an influence on bone mineral metabolism in growing goats and sheep. Twelve growing goats and sheep were divided into two groups. The two control groups received 6.1 g calcium/day (nG) and 6.7 g calcium/day (nS) for goat and sheep respectively. The other two groups were fed 17.7 g calcium/day (hG) and 18.5 g calcium/day (hS). Blood samples were taken 2, 4, 5 and 6 weeks after the start of the experiment. In serum Ca and vitamin D were determined and bone metabolism was measured using crosslinked carboxyterminal telopeptide of type I collagen (ICTP), crosslaps, bone-specific alkaline phosphatase and osteocalcin (OC). Bone mineral density (BMD) was quantified using quantitative computed tomography. Bone resorption marker (ICTP) concentrations were significantly different between both groups control sheep/control goat and hS/hG, but no significant differences were evident in the different feeding groups within one species. OC concentrations showed a similar course to ICTP. The goats had significantly higher concentrations compared with sheep. The 1,25 dihydroxyvitamin D (VITD) concentrations in both hCa groups were significantly lower than in the control groups. BMD increased in the hCa groups compared with the control groups with the time, but significant differences were only evident in sheep in week 2. The hCa diet did not induce differences between the groups within one species for all bone markers. The control Ca diet seems to improve the active Ca absorption via VITD whereas the hCa diet leads to a higher amount of Ca apparently digested. Higher BMD was only observed in group hS compared with nS. PMID- 15787982 TI - Influence of phytase added to a vegetarian diet on bone metabolism in pregnant and lactating sows. AB - The purpose of the study was to find out if the supplementation of phytase to a diet of gestating and lactating sows has any effects on performance and bone parameters of the animals. Forty primiparous gilts were assigned into four groups: group A with phytase [4.2 g total phosphorus (P)/kg (gestation) and 4.5 g total P/kg (lactation)], group B without phytase (with phytase supplementation in diet for rearing) and same P content as group A, group C without phytase and higher P contents [5.0 g total P/kg (gestation) and 5.5 g total P/kg (lactation)] and group D with the same diet as group B (no phytase during the rearing). A 6 phytase was used in this trial (750 FTU/kg diet). The four diets were fed during gestation and lactation. Faeces were collected to determine apparent digestibility of minerals. Blood samples were taken to analyse minerals and bone markers. After weaning the sows were slaughtered and the bones of one hind leg were prepared to measure bone mineral density (BMD) and bone mineral content (BMC) of the tibia. Bone ash and mineral content of the phalanx III were determined. Mean P concentrations in serum decreased during gestation and lactation. But there were no significant differences between the groups. Bone formation marker bone-specific alkaline phosphatase decreased at the beginning of lactation whereas bone resorption marker serum crosslaps increased. The BMD and BMC of the tibia were slightly higher in the groups fed higher concentrations of P and phytase. The ash and mineral contents of the phalanx were the highest for the group fed the highest concentration of P. The apparent digestibility of P increased during gestation mostly in group A (57%--> 69%). In conclusion, high P content and addition of phytase to the diet induced a slightly higher ash content of the bones. It is of high importance, that sows during gestation absorb enough P, to avoid lamenesses and sudden fractures. As not many studies with phytase have been performed during gestation and lactation in sows yet, we can recommend, that phytase as supplement can be used to keep P in the diet at a lower level without negative consequences for bone health. PMID- 15787983 TI - Effects of different phosphorus sources in the diet on bone composition and stability (breaking strength) in broilers. AB - In two fattening trials (in each 100 broilers kept in four groups with 25 animals) as well as in a balance trial (four groups with four broilers in a group) the effects of inorganic phosphorus sources [monocalcium phosphate (MCP), dicalcium phosphate (dihydrate; DCP) and defluorinated phosphate (DFP)] in broiler diets were examined. The four diets contained up to 9 g calcium and 6 g phosphorus per kg and comparable energy and nutrient contents. Controls were fed a commercial diet with Ca-Na-phosphate as inorganic phosphorus source supplemented by phytase. In both fattening trials body weight gain, feed consumption and feed conversion were proved as well as the calcium and phosphorus levels in serum, the breaking strength of tibia or humerus and the femur mineralization (ash content in the fat free dry matter). Furthermore, in the balance trial the retention of calcium and phosphorus was determined by calculation (intake minus excretion) as well as by analysis of body composition. On a high performance level (that was only slightly influenced by the different treatments), the addition of DFP resulted in significantly reduced phosphorus availability (estimated by analysis of the whole carcass: control/MCP/DCP/DFP: 48.6/46.0/45.7/35.5%). The significantly reduced phosphorus level in serum (1.77 +/- 0.20/1.77 +/- 0.24/1.73 +/- 0.28 1.34 +/- 0.33 mmol/l) indicates the lower phosphorus retention in broilers given DFP. Furthermore, the crude ash content (582 +/- 17.6/580 +/- 18.6/563 +/- 15.2/547 +/- 29.7 g/kg fat free DM) and the breaking strength of bones (in right tibia in trial 2: 232 +/- 82.4/227 +/- 51.5/232 +/- 41.7/196 +/- 655 N) were lowest when given DFP. For diagnostic purposes it is of special interest that the phosphorus levels in the serum reflected markedly the different concentrations of available phosphorus in the diet. PMID- 15787984 TI - Effects of draught load exercise and training on calcium homeostasis in horses. AB - This study was conducted to investigate the effects of draught load exercise on calcium (Ca) homeostasis in young horses. Five 2-year-old untrained Standardbred horses were studied in a 4-month training programme. All exercise workouts were performed on a treadmill at a 6% incline and with a constant draught load of 40 kg (0.44 kN). The training programme started with a standardized exercise test (SET 1; six incremental steps of 5 min duration each, first step 1.38 m/s, stepwise increase by 0.56 m/s). A training programme was then initiated which consisted of low-speed exercise sessions (LSE; constant velocity at 1.67 m/s for 60 min, 48 training sessions in total). After the 16th and 48th LSE sessions, SETs (SET 2: middle of training period, SET 3: finishing training period) were performed again under the identical test protocol of SET 1. Blood samples for blood lactate, plasma total Ca, blood ionized calcium (Ca(2+)), blood pH, plasma inorganic phosphorus (P(i)) and plasma intact parathyroid hormone (PTH) were collected before, during and after SETs, and before and after the first, 16th, 32nd and 48th LSE sessions. During SETs there was a decrease in ionized Ca(2+) and a rise in lactate, P(i) and intact PTH. The LSEs resulted in an increase in pH and P(i), whereas lactate, ionized Ca(2+), total Ca and intact PTH were not affected. No changes in Ca metabolism were detected in the course of training. Results of this study suggest that the type of exercise influences Ca homeostasis and intact PTH response, but that these effects are not influenced in the course of the training period. PMID- 15787985 TI - Metabolic responses to oral tryptophan supplementation before exercise in horses. AB - This study was conducted to evaluate the effects of oral tryptophan (Trp) supplementation on exercise capacity and metabolic responses in horses. Three horses had to perform an exercise test: a 15-min warm-up followed by a 60-min walk (1.7 m/s, W1), a 10-min trot (3.1 m/s, T1), a second 60-min walk (1.7 m/s, W2), a second 10-min trot (3.1 m/s, T2) and a final 30-min walk (1.7 m/s, W3) until the horses were unwilling to continue. The horses exercised on a treadmill at a 6% incline and with a constant draught load of 40 kg (0.44 kN). Two hours before exercise horses were given 50 g Trp (9.8-10.7 g Trp/100 kg BW) by nasogastric tube. A control exercise test was conducted without Trp. During the control test, one horse was able to finish the final 30-min walk (W3), whereas two horses finished W3 after Trp administration. Higher plasma Trp levels after Trp administration did not change significantly during exercise (Trp: start exercise, 524 +/- 41 micromol/l; end exercise 547 +/- 20 micromol/l; control: start exercise, 70 +/- 10 micromol/l; end exercise, 58 +/- 21 micromol/l). After Trp supplementation, blood lactate concentrations were significantly lower after the first and second trotting periods. Free fatty acids in plasma increased during exercise without any treatment-related differences. Although experimental plasma Trp levels were seven times higher than the control levels, Trp supplementation had no effect on exercise performance and metabolic responses to draught load exercise. PMID- 15787986 TI - Amino acid concentrations in blood serum of horses performing long lasting low intensity exercise. AB - The aim of this work was to evaluate the changes in the concentrations, after two rides different for distance covered, of different amino acids in endurance horses. Blood samples have been collected from horses just before the start, at the top of a steep slope (819 m difference in height) and just at the end of a 32 km endurance ride. A second group, competing in a 72 km endurance ride, has also been sampled immediately before and after the race. In serum samples, the concentrations of alanine, arginine, asparagine, glycine, isoleucine, histidine, leucine, lysine, methionine, ornithine, phenylalanine, tryptophan, tyrosine and valine have been measured by high-performance liquid chromatography (HPLC). anova and t-test have been used to study the differences in the concentrations of the amino acids. The pre-ride concentrations of the free amino acids were different between the two races, except for methionine and leucine. Differences between start and end race have been found for both groups for all the considered parameters except asparagine, isoleucine, leucine and lysine for the 72 km ride. Increases have been recorded for the shorter and decreases for the longer ride in the blood serum concentrations. Significant increases have also been found between the starting sampling and the second, at the top of the slope, only for alanine, arginine, asparagines, phenylalanine and lysine. The ride length has a significant impact on blood serum amino acids mobilization and uptake; in the shorter race the increases stand only for mobilization, whereas in the longer the decrease can be considered the effect of the onset of the amino acids catabolism. PMID- 15787987 TI - Colostrum management in calves: effects of drenching vs. bottle feeding. AB - The objective was to examine whether the administration of colostrum by a drencher is a safe method to achieve satisfying immunoglobulin concentrations in newborn Holstein Friesian (HF)-calves. Twenty-one calves were fed 1 h postnatum with 2 l colostrum from their dam using a nipple bottle (group I); 15 calves were drenched with 4 l colostrum from their dam 1 h postnatum (group II). Thereafter, all calves were fed milk replacer exclusively. Serum concentration of total immunoglobulin [analysed by Sandwich-enzyme-linked immunosorbent assay (ELISA)] 24 h postnatum was higher in drenched calves (25.2 g/l; 12.8/52.2; median and 10/90 percentiles) compared with bottle-fed calves (14.1 g/l; 9.2/24.7). To characterize the kinetics of immunoglobulin absorption, 17 blood samples were taken frequently within 72 h postnatum using a jugular catheter in five calves fed 2 l colostrum by nipple bottle and five calves drenched with 4 l colostrum 1 h postnatum. A slightly delayed increase of serum immunoglobulin concentration (approximately 3 h) was obvious in drenched calves compared with bottle-fed calves, but drenched calves reached significantly higher immunoglobulin concentrations compared with bottle-fed calves. It is concluded that the proper application of colostrum by a drencher is a useful method for adequate colostral supply in newborn calves. The failure of the oesophageal groove reflex in drenched calves caused no clinical consequences. PMID- 15787988 TI - Site and extent of starch degradation in the dairy cow - a comparison between in vivo, in situ and in vitro measurements. AB - Prediction of the supply of glycogenic precursors to dairy cows and the site of degradation of wheat, maize and potato starch (PS) were determined in an in vivo experiment and the results were compared with data obtained from experiments involving in situ nylon bag and in vitro gas production techniques. In a Latin square design experiment four lactating dairy cows fitted with a rumen cannula and T-piece cannulae in the duodenum and terminal ileum, received either a low starch control diet or diets in which sugar beet pulp in the concentrate mixture had been replaced by wheat, maize or PS. Starch from the different sources was almost completely degraded in the total gastrointestinal tract. For all starches, the rumen was the main site of degradation in vivo. No digestion of PS in the small intestine was observed. In situ results suggested that 14% of wheat starch (WS), 47% of maize starch and 34% of PS escaped rumen fermentation. According to the gas production technique WS ferments quickest and potato slowest. PS had a low degradability during the first 8 (gas production) to 11 (in situ) h. However, according to both in vitro and in vivo measurements rumen degradability of PS was high. The results suggest that in situ and in vitro techniques should be performed in animals that have adapted to starch source to provide a more accurate simulation of the in vivo situation. PMID- 15787989 TI - In situ ruminal degradability and intestinal digestion of raw and extruded legume seeds and soya bean meal protein. AB - An experiment was performed to evaluate the effect of extrusion and carbohydrate addition on rumen degradation and intestinal digestion of raw legume seeds and solvent extracted soya bean meal (SBM) protein. Whole soya beans (WSB) without or with maize added (75:25) (WSB-M), peas, lupins and SBM were extruded at 140 degrees C. Protein rumen degradation and intestinal digestibility of unprocessed and extruded protein sources were measured by in sacco and mobile bag procedures, respectively, in two dairy cows cannulated in rumen and duodenum. Between 12 and 15 polyester bags with 4 g of each protein source were incubated in rumen for 12 h and the residues, pooled by feed, were introduced into the duodenum in small nylon bags after pre-incubation in a pepsin solution, and recovered from faeces the day after. Extrusion significantly (p < 0.001) reduced N degradation of all protein sources, from 98.1%, 91.6%, 90.5% and 64.8% to 53.1%, 73.8%, 70.3% and 44.2% for peas, lupins, WSB and SBM respectively. The addition of maize to WSB strengthened the effect of extrusion on rumen N degradation, from 88.2% to 52.6%. Residues from rumen incubation of extruded feeds showed a higher (p < 0.001) intestinal N digestibility except for SBM (87.0%, 82.9%, 66.3%, 85.0% and 97.2%, and 99.1%, 95.8%, 96.8%, 97.8% and 98.7%, respectively, for non-extruded and extruded, peas, lupins, WSB, WSB-M and SBM). In conclusion, the extrusion of studied legume seeds and SBM promotes a clear and significant increase of their metabolizable protein value, particularly in peas, and the inclusion of a source of carbohydrates before extrusion increase this response. PMID- 15787990 TI - Investigations on the effects of Ca-soap of linseed oil on rumen fermentation in sheep and on milk composition of goats. AB - Six rumen-cannulated wethers were fed by a diet composed of alfalfa hay and concentrate and supplemented by 75 g Ca-soap of linseed oil (5.4% in dry matter, DM) daily. A model trial was performed to detect the effects of the Ca-soap on rumen fermentation parameters and on fibre digestion. Approximately 3 h after feeding Ca-soap, ratio of C2:C3 decreased (from 4.33 to 4.02) and the production of i- and n-butyrate and i- and n-valeriate increased by 28, 5.3, 11.76% and 6.80% respectively. Total volatile fatty acid concentration in rumen fluid did not change (126.1 vs. 126.4 mm) as a result of Ca-soap supplementation. The in vitro trial showed no detrimental influence of Ca-soap on the acid detergent fibre (ADF) degradation. Using feed samples containing Ca-soap to be incubated in tubes, ADF digestion proved to be significantly higher (p < 0.001). Approximately 14 goats (Saanen breed, 30-70 days in lactation) were used to test the effects of Ca-soap on milk composition. Their ration contained alfalfa hay, millet straw and a concentrate. In the experimental group (seven goats) the diet was supplemented with Ca-soap of linseed oil (75 g/animal/day). The milk composition was changed (slightly reduced solid content, sometimes significantly reduced milk fat contents), when Ca-soap was included in the diet of lactating goats. PMID- 15787991 TI - Beet-vinasse (condensed molasses solubles) as an ingredient in diets for cattle and pigs - nutritive value and limitations. AB - Sugar beet vinasse, originating as the condensed molasses residue from yeast production, was fed to bulls and pigs to determine the feed value. It contained, per kg dry matter (DM content 66.8%), 293 g crude protein, 305 g N-free extract (therefrom 34.4 g total sugar), and 395 g crude ash. The experiment was carried out with three fattening bulls (German Holstein) and four adult pigs (Gottingen Minipig). In bulls, two balance trials lasting 20 days each (10 days adaptation; 10 days collection) were conducted in which subsequently a basal ration without and with vinasse (14% in DM) was fed. In pigs, the experiment consisted of three 12-day balance trials with the final 5 days as collection period. In the first trial, pigs received a basal diet without vinasse and in the following, a mixed diet with 16 and 43% vinasse (in DM) respectively. Digestibilities of components in vinasse in bulls were: organic matter 73.5%, crude protein 72.6%, and N-free extract 52.3%. In bulls, no influence on faeces quality could be detected while feeding 14% vinasse in DM. While feeding 16% vinasse in DM to pigs, the digestibility of organic matter amounted to 72.3% (crude protein 71.8%; NfE 74.6%). Feeding higher amounts of vinasse (43% in DM) in the diet of pigs, digestibilities were reduced (organic matter: 61.6%), which might be a result of a reduced retention time of the chyme as a result of an osmotically driven diarrhoea because of unexpectedly high sulphate (136 g/kg DM) content in the vinasse. It is concluded that because of organic matter digestibility >70% for cattle and pigs, the use of vinasse in these species in small proportions is reasonable. PMID- 15787992 TI - Yeast (different sources and levels) as protein source in diets of reared piglets: effects on protein digestibility and N-metabolism. AB - The aim of this study was to examine the feeding value of different yeasts as a substitute for soya bean meal, the main protein source in diets of weaned piglets. Tested two yeasts were already available on the market, Saccharomyces cerevisiae and Kluyveromyces lactis (beer and milk yeast), which replaced 40% of the soya bean meal in the diets. Furthermore, a yeast (Kluyveromyces fragilis) grown on whey, a side-product of cheese production, was used in increasing concentrations in the diets, so that increasing amounts of the soya bean meal (20%, 40% and 60%) could be replaced. As proved in these experiments, a replacement of 60% of the soya protein with whey yeast protein had positive effects on the performances (daily weight gain) and on the N-metabolism and did not have negative effects on the health or the faeces consistency. The whey yeast stands out because of its high protein quality (N-digestibility and N-retention). Furthermore, the replacement of soya bean meal with highly digestible yeasts is welcomed under the aspect of animal health, because of the reduction of anti nutritive soya components (stachyose, glycinin) in diets of weaned piglets. The controlled production conditions of the yeasts result in a high feed safety; in addition, the yeast as an end-of-pipe-product is a resource conserving and valuable feed. A main stimulus for the use of yeasts, however, in a food production controlled by economic standpoints, is their price and the costs of other competing feeds. PMID- 15787993 TI - Relationship between total transit time and faecal quality in adult dogs differing in body size. AB - Fed the same diet, large and giant-breed dogs have higher faecal moisture and increased frequency of soft stools than small ones. This could be the result of physiological differences, such as a different gastrointestinal transit time. In this study, we have correlated mean total transit time (MTT) with body size and faecal consistency in dogs varying in body size. Fifty dogs from 13 different breeds were used, from a Dachshund to a Great Dane. The MTT was determined using coloured plastic beads [Cummings and Wiggins, Gut, Vol. 17 (1976), p. 219], and faecal consistency was scored daily during the study. We confirmed the strong correlation between height at the shoulder (body size) and faecal score (r = 0.76; p < 0.0001). The MTT increased with body size, from 22 h for a Miniature Poodle to 59 h for a Giant Schnauzer. We found significant positive correlations (p < 0.0001) between MTT and body size as well as faecal scores (r = 0.71 and 0.70 respectively). In the present study, we observed an effect of body size on MTT. In our 50 healthy dogs a longer MTT was related to a poorer faecal quality. Previous studies reported no relationship between body size and the upper gastrointestinal transit time in healthy dogs. So, we hypothesized that body size would mainly affect colonic transit time and that a longer colonic residence time would be related to a poorer faecal quality by promoting fermentation activity. PMID- 15787995 TI - Comparison of feed preference and digestion of three different commercial diets for cats and ferrets. AB - Diet preference and digestibility experiments were conducted using a total of 10 cats and 10 ferrets. The composition of the three different kinds of dry cat feed was as follows (each data are given in dry matter, DM): (i) normal diet (N): 95.3% DM, 33.7% crude protein (CP), 20.4% ether extract (EE), 37.6% nitrogen-free extract (NFE); (ii) 'light diet' (L): 94.2% DM, 31.6% CP, 10.7% EE, 52.2% NFE; (iii) 'veterinary diet' (D): 94.57% DM, 38.7% CP, 9.6% EE, 47.2% NFE. During the period of the preference test, the average daily dry matter intake (calculated with the mean of the three diets: 94.7% DM) was 98.0, 15.0 and 16.7 g DM in cats and 25.0, 7.3 and 8.1 g DM in ferrets. The preference rates of the three different diets, expressed in percentage of their total consumption, were as follows: 60.4% N (54.4 g DM), 12.4% L (12.1 g DM) and 27.2% D (26.6 g DM) in cats, and 46.2% N (11.6 g DM), 29.9% L (7.5 g DM) and 23.9% D (6.0 g DM) in ferrets. This indicates that cats and ferrets have a clear preference for diets of higher fat content. In all three diets, the digestibility of CP was significantly (p < 0.05) lower (70.1 +/- 5.4 vs. 75.9 +/- 5.8) while that of EE was significantly (p < 0.05) higher (95.6 +/- 1.5 vs. 89.4 +/- 5.3) in ferrets than in cats. The average digestible/metabolizable energy (DE/ME) ratio of feeds turned to be 95.6% for cats and 90.6% for the ferrets. From the data one can conclude that the ferret cannot be used as a model animal for cats either for preference or digestibility studies. PMID- 15787994 TI - Energy evaluation of extruded compound foods for dogs by near-infrared spectroscopy. AB - Near-infrared reflectance spectroscopy (NIRS) was used to predict the chemical composition, apparent digestibility and digestible nutrients and energy content of commercial extruded compound foods for dogs. Fifty-six foods of known chemical composition and in vivo apparent digestibility were analysed overall and 51 foods were used to predict gross energy digestibility and digestible energy content. Modified partial least square calibration models were developed for organic matter (OM), crude protein (CP), ether extract (EE), crude fibre (CF), nitrogen free extracts (NFE) and gross energy (GE) content, the apparent digestibility (OMD, CPD, EED, NFED and GED) and the digestible nutrient and energy content (DOM, DCP, DEE, DNFE and DE) of foods. The calibration equations obtained were evaluated by the standard error and the determination coefficient of cross validation. The cross-validation coefficients of determination (R) were 0.61, 0.99, 0.91, 0.96, 0.94 and 0.92 for OM, CP, EE, CF, NFE and GE, the corresponding standard error of cross-validation (SECV) being 5.80, 3.51, 13.35, 3.64 and 16.95 g/kg dry matter (DM) and 0.29 MJ/kg DM respectively. The prediction of apparent digestibility was slightly less accurate, but NIRS prediction of digestible nutrient (g/kg DM) and DE (MJ/kg DM) gave satisfactory results, with high R (0.93, 0.97, 0.93, 0.83 and 0.93 for DOM, DCP, DEE, DNFE and DE respectively) and relatively low SECV (11.55, 6.85, 12.14 and 22.98 g/kg DM and 0.47 MJ/kg DM). It is concluded that the precision of NIRS in predicting the energy value of compound extruded foods for dogs is similar or better than by proximate analysis, as well as being faster and more accurate. PMID- 15787996 TI - Essential fatty acids supplementation in different-stage atopic dogs fed on a controlled diet. AB - The aim of this trial was to evaluate the effects of polyunsaturated fatty acid (PUFA) supplementation in different-stages atopic dogs fed on a controlled diet. Twenty-two non-seasonal atopic dogs of different breeds and ages were included in the 2-month trial. All the patients were given an essential fatty acid (EFA) supplementation [17 mg/kg eicosapentaenoic acid (EPA) + 5 mg/kg docosahexaenoic acid (DHA) + 35 mg/kg gammalinolenic acid (GLA)], the global (diet + supplementation) omega-6 to omega-3 ratio was 5.5-1. Two groups of dogs were considered: group A 'pre-immunotherapy' (15 cases) included dogs with early stages atopy, which had not been submitted to any treatment yet; group B 'post immunotherapy' (seven cases) included dogs with chronic atopy immunotherapy non responsive. Clinical evaluations were performed at the beginning, on day 30 and at the end of the trial. Blood serum fatty acids profile was determined at the beginning and at the end of the study. Better clinical results were obtained in group A, a great difference was found between the two groups on pruritus score. Serum arachidonic acid (AA) was significantly lower at the end of the trial in group A while GLA was significantly higher in group B. We hypothesized that different-stages atopic dogs could have different response to EFA supplementation, maybe because of a different fatty acids metabolism. Early stages cases seem to be more responsive to EFA supplementation. PMID- 15787997 TI - The effect of oligofructose on urea metabolism and faecal odour components in cats. AB - The effect on urea metabolism by the supplementation of oligofructose to a reduced protein diet was evaluated in cats by the use of labelled urea. The effect on faecal odour was also evaluated. Four cats were tested in a crossover study with two treatments: control and fructo-oligosaccharide (FOS). The FOS was supplemented at 3.11% on dry matter (DM) basis to a reduced protein diet (28.9% DM). After an adaptation period of 3 weeks, faeces and urine were collected during a 5-day collection period. Fresh faecal samples were collected for determination of odour components. On the first day of the collection period, labelled urea was injected subcutaneously. Urine production was estimated by potassium excretion. The fresh faecal samples were incubated in an air-closed recipient with two solid-phase micro extraction (SPME) fibres to bind the produced sulphur (S)-containing components. The reduced protein diet decreased plasma urea concentration but FOS supplementation had no effect. The tendency for a higher faecal output by FOS supplementation was the consequence of both an increased moisture content and faecal DM production. Supplementation of FOS showed tendencies to increase total faecal nitrogen (N) excretion and faecal (15)N excretion and tended to decrease urinary (15)N excretion. Twenty-seven different odour components were detected but were not affected by FOS supplementation. PMID- 15787998 TI - Comparison of the nutrient analysis and caloric density of 30 commercial seed mixtures (in toto and dehulled) with 27 commercial diets for parrots. AB - In this paper, an overview is given of the composition of 30 commercially available parrot seed mixtures. As parrots dehull the seeds, the analysis of the total seed mixture tends to differ from that of the ingested feed. Statistical evaluation and comparison of the dehulled seeds vs. the whole seeds indicates that most parrot species are fed a diet rich in fat (31.7 +/- 13.1% crude fat) and energy (22.4 +/- 2.9 MJ ME/kg). As the analysis of the total seed mixtures underestimates fat and energy content of the ingested feed, it is suggested that researchers, bird nutritionists and bird food producers should calculate diets based on the analysis of the dehulled seeds. Finally, the calculated data were compared with the composition of formulated pelleted/extruded diets on the market. These data indicate that the energy density of most diets (15.6 +/- 1.4 MJ ME/kg) is far below the energy density of common seed mixtures. PMID- 15787999 TI - Investigations in pet birds (Agapornis spp.) fed different vitamin K contents in the diet. AB - In the past many discussions about possible toxic effects of vitamin K(3) fed to pet birds arose frequently, and were published also in magazines for pet bird fanciers, in the internet as well as in veterinary journals. Therefore, the aim of this study was an evaluation of effects of different dosages of vitamin K(3) on birds' health when given orally for a longer period. These investigations were carried out with adult lovebirds (Agapornis spp.) fed a pelleted diet with different levels of vitamin K(3) (menadione-sodium-bisulphite): control group 0 mg, group V1 20 mg and group V2 200 mg/kg diet dry matter. General condition and well being of the lovebirds were checked daily. Body weight gains as well as feed and water intakes were examined once a week. Every 2-month blood samples of each lovebird were collected and analysed. After a period of 6 and 10 months, respectively, four birds of each group were necropsied in order to carry out a pathological and histological examination. In general, the behaviour, feed and water intakes as well as quality of excreta were not influenced by ingestion of diets with different levels of vitamin K(3). All variations were in physiological ranges. Individuals of all groups showed positive body weight gains and an active reproduction status. However, the best body mass (BM) development and egg laying activity could be observed in lovebirds of group V2 with the highest vitamin K(3) supplementation. In the haemotogram some time-depending variations could be observed; however, a systematic influence of vitamin K(3) could not be determined in any group or at any time. All analysed biochemical values in plasma and the activities of enzymes were within normal ranges. Only few birds of every group showed aberrant histological findings, but none of these could be related to the vitamin K(3) intake. Moreover, no forced accumulation of vitamin K(3) in the liver depending on vitamin K(3) intake was found. This result suggests a rapid metabolism of the absorbed vitamin K(3). All in all, the application of pelleted diets with addition of 20 or 200 mg vitamin K(3)/kg diet over a period of several months did not affect pet birds' health. Given these results, any doubts about the compatibility of usual doses of vitamin K(3) in diets for lovebirds must be considered as absolutely groundless. PMID- 15788001 TI - Neutrophil serine proteases: potential key regulators of cell signalling during inflammation. AB - The serine proteases cathepsin G, human leucocyte elastase and proteinase 3 are major contents of neutrophils and are released at sites of inflammation. The common picture of their function was that they do not degrade extracellular proteins specifically. Recent studies provided evidence that these proteases are able to activate specifically pro-inflammatory cytokines and lead to the activation of different receptors. Neutrophil serine proteases might therefore be important regulators of inflammatory processes and are interesting targets for new therapeutic approaches against inflammatory disorders. This review summarizes the current knowledge on the regulation of cell signalling by neutrophil serine proteases. PMID- 15788000 TI - Studies on digestive physiology and feed digestibilities in captive Indian rhinoceros (Rhinoceros unicornis). AB - We performed intake, digestibility and ingesta passage studies in 11 Indian rhinoceroses (Rhinoceros unicornis) from four zoological institutions, using total faecal collection for the quantification of faecal output. The regularly fed zoo ration of roughage and concentrates (ration RC) and a roughage-only ration (ration R) were used; the roughage source differed between the facilities and comprised grass hay, grass silage, straw and lucerne hay. Dry matter intake ranged between 0.8 and 1.3% of body weight on ration RC and 0.5-1.2% on ration R. Digestibility coefficients achieved were similar to those reported for horses on diets of comparable composition. Endogenous losses as determined by linear regression analysis were within the range reported for horses. Measurements of faecal volatile fatty acids, faecal lactate and faecal pH also showed similarity to similar measurements in horses. The mean retention times of fluids (Co-EDTA) and particles (Cr-mordanted fibre <2 mm) in the whole gastrointestinal tract averaged 42 and 61 h, respectively, and were the longest ever recorded in a monogastric ungulate with this marker system. The results suggest that the horse is a useful model animal for designing diets for Indian rhinoceroses. Why digestive parameters are similar between these species in spite of enormous differences in body weight and retention times remains to be answered. PMID- 15788002 TI - Dietary habits after myocardial infarction - results from a cross-sectional study. AB - OBJECTIVE: Comparing habitual nutrient intakes in persons with a history of acute myocardial infarction (AMI), and age-matched controls. Design. Cross-sectional study. Subjects. Men and women (525 cases and 1890 matched controls), aged 47-73 years, of the population-based Malmo Diet and Cancer cohort. METHODS: Nutrient intakes were assessed by a validated modified diet history method. Body fatness was assessed by bioimpedance analysis. Case ascertainment was provided by national and regional registries. Men and women were analysed separately. Median time since AMI was 5.5 years in men and 3.8 years in women. Cases reported lower energy intakes (EIs) than controls, despite having similar basal metabolic rates. After adjustment for total EI, both male and female cases had lower fat intake and higher intake of several micronutrients, such as ascorbic acid, folate, and vitamin E, than controls, the difference being largest in men. Most of the cases reporting dietary change quoted 'disease' as their main reason for change. They had lower EI and lower energy-adjusted intake of fat than other cases. CONCLUSIONS: Survivors of AMI reported dietary habits more in line with current recommendations, particularly those who afterwards reported having changed their dietary habits. The possible bias introduced by social desirability is discussed. PMID- 15788003 TI - Weight change and mortality: the Nord-Trondelag Health Study. AB - OBJECTIVES: The prevalence of obesity is increasing. Overweight and obese people have increased mortality compared with normal weight people. We investigated the effect of weight change on mortality. DESIGN: Prospective population study. SETTING: We utilized data from two large population-based health studies conducted in 1984-86 and 1995-97 respectively. Cox proportional hazards models were used to calculate mortality rate ratios (RRs) with 95% confidence intervals (CIs) between people with a stable weight and people who lost or gained weight. Subjects. Totally 20,542 men and 23,712 women aged 20 years or more, without cardiovascular disease or diabetes at the first survey and without a history of cancer at the second survey were followed up on all-cause mortality for 5 years after the second survey. RESULTS: We found no association between weight gain and mortality. People who lost weight had a higher total mortality rate compared with those who were weight stable [RR was 1.6 (95% CI: 1.4-1.8) in men and 1.7 (95% CI: 1.5-2.0) in women]. Similar associations were found for cardiovascular and noncardiovascular mortality. Additional analysis showed a linear increase in mortality rates across categories of weight loss for both men and women (P < 0.001). There was a statistically significant interaction between weight change and initial BMI, but only amongst men (P = 0.001). CONCLUSIONS: Weight loss, but not weight gain, was associated with increased mortality amongst men and women. Although underlying undiagnosed disease is the most plausible explanation for this finding, the similar associations found for total mortality, cardiovascular mortality, and noncardiovascular mortality makes the causal pathway somewhat enigmatic. PMID- 15788004 TI - Ultrasound imaging in diagnosis of superior mesenteric artery syndrome. AB - OBJECTIVES: We screened a cohort of subjects affected by various degree of dyspepsia to reveal if they presented a reduction of the aorto-mesenteric angle and to diagnose suspected cases of superior mesenteric artery (SMA) syndrome. DESIGN: Controlled, prospective, study. SETTING: Subjects were studied as outpatients. SUBJECTS: The study investigated a total of 3622 subjects referred to our department by their general practitioners for dyspepsia and/or abdominal pain. Interventions. Abdominal ultrasonography with power colour Doppler, gastroduodenoscopy, hypotonic duodenography, contrast-enhanced spiral computerized tomography were performed. MAIN OUTCOME MEASUREMENT AND RESULTS: Color Doppler revealed a significant reduction of the SMA angle in 29 of 950 subjects; gastroscopy showed duodenal compressive pulsation in 14 of 29 patients and X-ray revealed compression of the third segment of the duodenum in 28 of 29 patients. CT confirmed the presence of a reduced angle and various degrees of duodenal compression in all patients. Ultrasonography and CT examinations gave overlapping results (P > 0.05) in diagnosing pathological aorto-mesenteric angle. CONCLUSION: The authors believe that the incidence of reduced aorto-mesenteric angle and SMA syndrome might be underrated. Ultrasound power colour Doppler imaging is useful in epidemiological screening of reduced aorto-mesenteric angle to diagnose suspected cases of SMA syndrome. PMID- 15788005 TI - Venous thromboembolism prophylaxis in acutely ill medical patients: definite need for improvement. AB - AIM OF THE STUDY: To examine the frequency and adequacy of thromboprophylaxis in acutely ill medical patients hospitalized in eight Swiss medical hospitals. METHODS: A cross-sectional study of 1372 patients from eight Swiss hospitals was carried out. After exclusion of patients (275) given therapeutic anticoagulation, 1097 patients were audited. The adequacy of thromboprophylaxis was assessed by comparison with predefined explicit criteria. RESULTS: Of 1097 patients, 542 (49.4%) received thromboprophylaxis. According to the explicit criteria, 644 (58.7%) should have been on prophylaxis (P < 0.001, when compared with the rate observed). The rate of prevention differed widely between hospitals (from 29.4 to 88.6%) with no difference between teaching and nonteaching hospitals. According to the explicit criteria, a substantial proportion (44.9%) of the patients who should have been treated were not. Conversely, 41.3% of the patients were unnecessarily treated. CONCLUSIONS: Even though the appropriateness of the explicit criteria used could be challenged, our data suggest that the current practice is associated with important uncertainty leading to both overuse and underuse of thromboprophylaxis in patients hospitalized in medical wards. More efforts are urgently needed to develop new or endorse existing explicit, evidence based criteria and guidelines for thromboprophylaxis in this population of patients. PMID- 15788006 TI - Evidence of a dynamic aldosterone-independent distal tubular control of renal sodium excretion in compensated liver cirrhosis. AB - BACKGROUND AND AIM: In preascitic cirrhosis increased sodium retention occurs in kidney distal tubule in spite of normal aldosterone plasma levels. No clearance technique can dissect the respective contribution to sodium retention exerted by Henle's loop, distal convoluted tubule and collecting duct, so we evaluated proximal and distal tubular sodium handling in preascites during two manoeuvres that temporarily increase aldosterone secretion. METHODS: Ten patients with compensated cirrhosis and nine controls were studied in recumbency, during standing and after dopamine receptor blockade with metoclopramide through: 4 h renal clearances of sodium, potassium, lithium and creatinine; plasma levels of active renin and aldosterone. RESULTS: Whilst comparable in recumbency, aldosterone levels significantly rose during standing and after metoclopramide in both groups. In patients, dopaminergic blockade caused a fall of distal sodium delivery (P < 0.01) but urinary sodium excretion was unchanged because the reabsorbed fraction of distal sodium delivery also fell (P < 0.03). Cirrhotic patients showed the same findings in the passage from recumbency to standing. CONCLUSIONS: In preascitic cirrhosis, the distal tubular segments of the nephron are able to cope with decreases in tubular flow by reducing reabsorption at an aldosterone-independent site (possibly the loop of Henle). PMID- 15788007 TI - Tumour staging, morphology and p53 overexpression concur in predicting survival in hepatocellular carcinoma. AB - BACKGROUND/AIMS: The prognosis of hepatocellular carcinoma (HCC) on cirrhosis is hard to predict as it depends on tumour stage, underlying liver disease, type of treatment and, possibly, biological factors of the tumour itself. METHODS: We prospectively evaluated the survival of 91 consecutive patients with HCC on cirrhosis, diagnosed between January 1998 and December 1999. Clinical features and histological/biological aspects, including histotype, grade, p53 overexpression, cytoproliferation and apoptotic markers were analysed. RESULTS: Child-Pugh (P = 0.01), Okuda (P < 0.0001), Cancer of the Liver Italian Program (CLIP) staging (P < 0.0001) and type of treatment (P = 0.0001) were significantly related to survival. In the Cox model, CLIP staging was included as independent predictor of survival at step 1 (P < 0.0001) with Okuda at step 2 (P = 0.013). Amongst the biological factors, p53 overexpression and histotype were significantly related with survival (P = 0.0044 and 0.017 respectively). When clinical and biological variables were examined together in the Cox model, CLIP and Okuda were confirmed as being statistically related with survival (P < 0.0001 and =0.012) followed by histotype and p53 overexpression (P = 0.019 and 0.02). CONCLUSIONS: CLIP, Okuda, histotype and p53 overexpression are the strongest predictors of survival in this series of patients. These data confirm that staging of the tumour and underlying liver disease are strictly related to prognosis but support the concurrent role of clinical and biological factors in upgrading our capacity of predicting the fate of HCC patients. PMID- 15788008 TI - Fracture risk associated with systemic and topical corticosteroids. AB - OBJECTIVES: To study the fracture risk associated with the use of corticosteroids in any formulation and administration. DESIGN: Case-control study. SETTING: Community-based study in Denmark. Subjects. Cases were all subjects with any fracture sustained during the year 2000 (n = 124,655). For each case, three controls (n = 373,962) matched on age and gender were randomly drawn from the background population. Adjustments were made for concurrent diseases (lung diseases, rheumatic disorders), use of other drugs (inhaled bronchodilators), contacts to hospitals and general practitioners, and social variables. RESULTS: An increased risk of any fracture, hip, spine, and forearm fractures was present with use of more than 2.5 mg of prednisolone or equivalent orally per day. For inhaled corticosteroids a limited increase in the risk of any fracture was present in users of more than 7.5 mg prednisolone equivalents per day (equivalent to 1875 mug of budesonide per day). However, no increase in the risk of hip, spine or forearm fractures was present in users of inhaled corticosteroids. For other topical corticosteroids (dermal, rectal, nasal, local application in the mouth, the eyes or the ears) no increase in fracture risk could be demonstrated even at high doses after adjustment for confounders. CONCLUSIONS: Ingestion of more than 2.5 mg of oral prednisolone equivalents per day is associated with an increase in fracture risk. No increase is associated with inhaled corticosteroids except at daily dosages above 7.5 mg of prednisolone equivalents. No increase in fracture risk is associated with other forms of topical corticosteroids. PMID- 15788009 TI - Zoonotic transmission of Cryptococcus neoformans from a magpie to an immunocompetent patient. AB - We report a case of cryptococcal meningitis in an immunocompetent female patient with exposure to a pet magpie (Pica pica). Genetically indistinguishable isolates were cultured from the cerebrospinal fluid of the patient and excreta of the bird. Our data strongly suggest zoonotic transmission of Cryptococcus neoformans var. grubii from a magpie to this patient. PMID- 15788010 TI - Iron status and HFE mutations in first-degree relatives of patients with sporadic porphyria cutanea tarda, in a Mediterranean area. PMID- 15788014 TI - Preferring patient-reported to observer-reported outcomes substantially influences the results of the updated systematic review on placebos by Hrobjartsson and Gotzsche. PMID- 15788017 TI - Patient-centred outcomes in dietary research. AB - INTRODUCTION: Current health policy stresses the need to take patients' views into account in the provision of health care, including involvement in the process of care and determining the outcome(s) of treatment. It is unclear whether patients undergoing dietary treatment have been consulted on their views. METHODS: A literature review, searching electronic databases and hand searching dietetic journals, of dietary intervention research was carried out to identify if patients' views were sought in determining outcomes. RESULTS: Searching uncovered a total of 28 papers reporting the results of studies of dietary interventions where patients views were sought in the measurement of outcome, however little detail was given about patient(s) involvement and whether the use of the measures influenced future care. FUTURE WORK: Should address the issue of patient involvement in the determination of outcome and how this then influences future care. PMID- 15788018 TI - Dietary habits and children's family lives. AB - PURPOSE: To investigate associations between 'less healthy eating' and 'unhealthy snacking' at age 11, and family life (family structure, meals and maternal employment status) together with potential socio-economic confounders and gender. METHODS: Children participated in a school-based survey, questionnaires also being completed by parents. Analyses were based on those with complete data, weighted to account for bias in return of parental questionnaires (N = 2146). Data from a dietary inventory, questions on food choice and snacks were used to classify 'less healthy eating' and 'unhealthy snacking'. RESULTS: Less healthy eating' (57%) and 'unhealthy snacking' (32%) were associated with greater deprivation, fewer maternal qualifications and being male. Compared with children of full-time homemakers, the likelihood of 'less healthy eating' was reduced among those whose mothers worked part-time (this effect remaining after socio economic adjustment), full-time (effect removed after adjustment), or were unemployed, sick or disabled (effect emerging after adjustment). 'Unhealthy snacking' was not related to maternal employment, and neither measure was associated with family structure or daily meals. CONCLUSIONS: There was no evidence that family structure or meals were associated with children's diets, although maternal employment had a positive association. Contrasting with the size of these effects, relationships between diet and socio-economic status were strong. PMID- 15788019 TI - The CAFE computer program for nutritional analysis of the EPIC-Norfolk food frequency questionnaire and identification of extreme nutrient values. AB - BACKGROUND: Food frequency questionnaires (FFQs) are widely used in nutritional epidemiology but no papers detail the development of the supporting programs and nutritional databases. The principles involved in data collection, processing and treatment of the European Prospective Investigation into Cancer Study (EPIC) Norfolk Study FFQ and development of the structure and content of the Compositional Analyses from Frequency Estimates (CAFE) program for calculating nutrient intakes are described. Extreme nutrient values and derivation of cut points for data exclusion are also discussed. METHODS: The FFQ was sent to 25,637 participants. To exclude extreme (nonphysiological) outliers of nutrient intakes, the extreme top and bottom 0.5% of the ratio of energy intake to estimated basal metabolic rate was used. RESULTS: The CAFE computer program modifies nutrient intake according to specific fats used in food preparation and the amount of visible fat on meat. It incorporates different breakfast cereals and distinguishes between text for brands and types. After exclusion of outliers of nutrient intake some extreme values remained for energy, carbohydrate, protein, fat and alcohol, because of high reported frequencies of certain foods. CONCLUSIONS: The features of CAFE include flexible, updateable, databases and a novel method for matching text. The effect of extreme nutrient values on the accuracy of the FFQ in estimating diet in nutritional epidemiology should be investigated further. PMID- 15788020 TI - Food habits and appetite of elderly women at the time of a femoral neck fracture and after nutritional and anabolic support. AB - OBJECTIVE: To study appetite and food choices in lean elderly women at the time of a femoral neck fracture and after 6 months of nutritional and anabolic intervention. SUBJECTS AND METHODS: Forty-five nondemented women >70 years of age (mean +/- SD: 83 +/- 5 years) with a recent hip fracture and body mass index <24 kg/m2 (mean: 20.5 +/- 2.3) were interviewed about their appetite and dietary habits prior to fracture. The patients were randomized to treatment with a protein-rich liquid supplement (PR; 200 kcal and 20 g protein day(-1)) alone or in combination with nandrolone decanoate injections (PR/N) 25 mg i.m. every third week or to a control group (C). A second interview was conducted 6 months later. RESULTS: Reduced appetite before the fracture was reported by 60%. Half of the patients did not have dessert with any of their daily meals, one-third used low fat margarines and one-third drank water with their meals. The estimated mean daily energy intake was 6.4 +/- 1.2 MJ (1541 +/- 304 kcal) indicating that three of four subjects did not meet their energy needs. At 6 months, 40% reported reduced appetite. There was no difference in the change of appetite between the three randomized groups. Still, half of the subjects appeared to not meet their energy needs. Protein intake increased in the PR and PR/N groups, in contrast to the controls (P = 0.002). CONCLUSION: Reduced appetite and insufficient energy intake was recorded in lean elderly women with a femoral neck fracture. Nutritional supplementation alone or in combination with an anabolic steroid increased protein intake without adversely affecting appetite. PMID- 15788021 TI - Dietary strategies and uptake of reduced fat foods. AB - AIM: This research has sought to identify factors determining consumer uptake and rejection of reduced fat foods. Initial qualitative research implied that use of reduced fat products could be dependent upon which dietary health strategies people adopt. This analysis therefore explores the degree to which various dietary strategies predict uptake of reduced fat products. METHOD: Items were derived from prior qualitative findings and responses to relevant issues surveyed by interview (n = 1004) within selected retail outlets throughout Northern Ireland and England. RESULTS: Logistic regression analysis indicated that concern over health, drive to reduce calorie consumption, to increase intake of fish and poultry and fruit and vegetables consistently predicted consumption across a range of reduced fat products. These dietary strategies appeared to predict uptake of reduced fat foods to a greater degree than perceived taste or price. CONCLUSION: The findings imply that for reduced fat foods to achieve universal acceptance promotional messages need to be translated into commonly adopted dietary health strategies. PMID- 15788022 TI - Nutritional status and clinical outcomes of older patients in rehabilitation. AB - BACKGROUND: Malnutrition is associated with poor outcomes in older adults and those admitted to rehabilitation may be particularly at risk. Objective To assess the nutritional status and outcomes of older adults in rehabilitation. SUBJECTS AND METHODS: We recruited 133 adults > or = 65 years from consecutive rehabilitation admissions. Nutritional status was assessed using the mini nutritional assessment, body mass index (BMI) and corrected arm muscle area (CAMA). Outcomes measured included length of stay, admission to higher level care, function and quality of life (QOL). RESULTS: Sixty-two (47%) subjects were well nourished, 63 (47%) at risk of malnutrition and eight (6%) malnourished. Twenty-two (17%) and 27 (20%) were below the desirable reference values for BMI and CAMA respectively. Subjects at risk of malnutrition/malnourished had longer length of stay (P = 0.023) and were more likely to be admitted to higher level care (P < 0.05). These subjects also had poorer function on admission (P < 0.001) and 90 days (P = 0.002) and QOL on admission (P < 0.008) and 90 days (P = 0.001). Those with low CAMA were twice as likely to be admitted to higher level care (P < 0.05) and had poorer function at 90 days (P = 0.017). CONCLUSIONS: Over half our sample was identified as at risk of malnutrition or malnourished and this was associated with poorer clinical outcomes. PMID- 15788026 TI - Obesity management. PMID- 15788027 TI - Obesity management. PMID- 15788028 TI - Developing a visible critical mass. PMID- 15788029 TI - Psychosocial assessment of patients who attend an accident and emergency department with self-harm. AB - It has been recommended that all patients attending hospital accident and emergency departments in the United Kingdom (UK) with self-harm should be offered adequate psychosocial assessment to identify those at risk of completed suicide or repeated self-harm. This paper focuses on the extent to which this recommendation has been implemented in a large district general hospital in Wales, and compares the characteristics and hospital management of patients who received no specialist assessment with those who were assessed by a psychiatric liaison nurse or a psychiatrist. Data were routinely collected as part of the hospital annual audit procedure and confirm previous findings that guidelines for the general hospital management of self-harm patients have not been well implemented. Whilst we found some evidence that certain high risk patient groups were more likely to receive specialist assessment, this was far from universal. We conclude that if there is to be a reduction in the rate of suicide in Wales and the UK as a whole, it is crucial that services for people who self-harm are more appropriately targeted on patients who are at greatest risk. PMID- 15788030 TI - Organizing and delivering training for acute mental health services: a discussion paper. AB - Recent policy statements that address the quality of care provided by acute mental health services have highlighted an urgent need for specialist nurse education and training. However, examples of how to design and implement such training initiatives are sparse. Drawing on recent experience of developing an innovative training programme for acute psychiatric settings, this paper seeks to examine some of the key issues associated with current training provision for acute inpatient mental health workers. The methodological and practical concerns surrounding this type of initiative are discussed with the main aspects of programme content, service user participation, team training and organizational challenges being explored. Resulting from this work, several recommendations regarding the content, organization and delivery of future training initiatives are made. PMID- 15788031 TI - Aspects of the relationship between doctors and depressed patients that enhance satisfaction with primary care. AB - Depression will be the second most common threat to health by the end of the next decade. The incidence of depression in primary care is already high. This has serious implications for the types of services available, the range of personnel who deliver them and the extent to which patients are helped by the treatments provided. Research reveals that approximately two-thirds of patients with depression are treated with medication, although it is not known how effective medication is in the long term. It would appear that the relationship between the patient and prescriber is a highly significant factor in determining whether or not patients adhere to treatment regimens and to what extent they improve. This study used a qualitative approach to identify how patients treated with medication for their depression perceived the relationship with their prescribing clinician, what kinds of information and advice they received and what they wanted. Several aspects of the helpful relationship are identified, such as the characteristics and behaviour of the clinician, as well as the way in which information is imparted. The frequency of monitoring consultations and patients' perceptions of their importance were also examined. The issue of stigma, particularly self-stigma and what can be done to prevent it, is discussed. The paper concludes with suggestions for improving the quality of primary care for patients' prescribed medication for depression, and especially for making maximum use of the initial consultation. The main implication for mental health nurses is that central to all interventions for depression is the primacy of the relationship, without which, clients' belief in treatment is diminished. PMID- 15788032 TI - Emotional labour and stress within mental health nursing. AB - For many within the nursing profession, the work role involves a great deal of emotional work or 'emotional labour'. Such emotional work can be performed through 'surface acting' in which the individual simply feigns an appropriate emotion, or through 'deep acting' in which they actually try to feel the required emotion. The current study aims to aid understanding of the complex relationship between components of emotional labour and stress within the mental health nursing sector. Thirty-five mental health nurses completed questionnaires relating to a total of 122 nurse-patient interactions. Data were collected in relation to: (1) the duration and intensity of the interaction; (2) the variety of emotions expressed; (3) the degree of surface or deep acting the nurse performed; and (4) the perceived level of stress the interaction involved. Nurses also completed Daily Stress Indicators. Results suggest that: (1) emotional labour is positively correlated with both 'interaction stress' and daily stress levels; (2) the deeper the intensity of interactions and the more variety of emotions experienced, the more emotional labour was reported; and (3) surface acting was a more important predictor of emotional labour than deep acting. Implications for mental health nurses are outlined. PMID- 15788033 TI - Medicine taking decisions: schizophrenia in comparison to asthma and epilepsy. AB - This paper outlines a recent PhD study that explored factors underpinning decisions of people with schizophrenia regarding neuroleptic medicine taking. Relevant studies tend to be within the world view of psychiatry, resting on traditional concepts of compliance and insight. These studies examined schizophrenia outwith the context of other illnesses taking a snap shot view of medicine taking decision making, thus de-emphasizing its process. The chosen method of grounded theory (Glaser and Strauss 1967) overcomes these deficiencies. Stage 1 data arose from patient interviews, patients with asthma, epilepsy and schizophrenia contributed (n = 45), carers also sometimes contributed (n = 11), all three conditions being episodic disorders vulnerable to environmental stressors. Stage 2 develops from stage 1, mental health workers involved in the care of people with schizophrenia were interviewed, plus one asthma specialist and one epilepsy specialist (n = 16). Their views were compared to perspectives arising from the patient interviews and were included in the data, out of which an explanatory typology emerged. The three parts of the typology comprise the study's major categories and are labelled as: direct-reactive, deferential compliant, and active discerning and optimizing. Active discerning and optimizing consists of two stages; experimental-reflective and consolidation. The core category of responsive resolution contains themes which run through all major categories. PMID- 15788034 TI - Happy with your care? AB - With the recent rise of consumerism within health care, the views of service users are becoming increasingly important. The National Service Framework for Mental Health (DoH 1999) identifies assertive outreach as a strategy to engage people with severe and enduring mental illness, who traditionally lose contact with services. Assessing the views of service users is not a new strategy for service evaluation, but in relation to assertive outreach in the United Kingdom is relatively untested. A postal client satisfaction survey was undertaken as a means of enquiry, in order to ascertain the effectiveness of the Preston Assertive Outreach service from a service user perspective after its first year of operation. The team offers a service based upon the PACT (Program of Assertive Community Treatment) model. An evaluation study of 36 questionnaires produced a return rate of 56% (21 returns). The results showed a fairly high level of client satisfaction with the service provided. Client involvement in treatment was the area of service evaluated least favourably by the service users, with General satisfaction showing the highest level of client satisfaction. From the outcome of the study it appears there are potential benefits in utilizing feedback from service users' to inform future service provision. PMID- 15788035 TI - Substance use and young people: the potential of technology. AB - There is widespread concern about alcohol and drug use and abuse among young people in the United Kingdom, Europe and the United States. Evaluations of current drug and alcohol education approaches have mixed findings and some methodological difficulties. This paper reports on exploratory research to explore the potential of technology to provide information and support to young people regarding substance use and abuse. Eight focus groups were conducted with young people (n = 78) and three with key informants (n = 22). The findings revealed that technology has potential to provide information on this topic, and that young people are critical of some of the more traditional methods of provision. The young people were prepared to experiment with technology, felt competent to do so, and made suggestions of information they would like. These findings are discussed and recommendations made for future research in the area. PMID- 15788036 TI - Culture shift: carer empowerment and cooperative inquiry. AB - Government policy has directed local services to address the needs of carers as a way of maintaining care in the community. This study was initiated to enable carers to develop an information pack based upon their identified needs. Cooperative inquiry was the method used to ensure full participation of the carers. Group meetings were already in existence through a charity organization who provides a carers support network. The first author participated in a number of carers group meetings. Cooperative inquiry was used to clarify a number of themes identified and reflective cycles ensured that those themes remained relevant. It was found that carers did want to be involved in their relative's care, not as passive recipients but as collaborative care providers. To do this they need to be fully informed of the processes of care provision. Carers need information that is relevant, easily accessible and obtainable in varying degrees of comprehension. This study suggests that a culture shift within mental health nursing is necessary if professionals are to recognize that a perceived lack of support may lead to a breakdown in relationships between the carer, the person being cared for and the professionals. PMID- 15788037 TI - A framework for good practice in interagency interventions with cases of self neglect. AB - Self-neglect is probably more common than has generally been thought and gives rise to a number of difficult conceptual, identification and intervention problems for health and social care workers. These patient management issues are compounded by the lack of any evidence-base for practice which nurses can draw on when faced with managing people who self-neglect. Nurses working with self neglecters operate in an interdisciplinary context which includes professional groups, such as environmental health officers, who do not normally figure within health and social care teams. In the absence of such an evidence-base, a pragmatic solution in the form of a framework for interagency practice has been proposed. The background underpinning this framework, including a research study of housing in self-neglect and interagency responses, is briefly outlined in this paper. PMID- 15788038 TI - A survey of trainees' opinions and current clinical practice after behaviour therapy training. AB - The widespread use of cognitive behaviour therapy principles in psychiatric/mental settings has been an ongoing feature of health and social care since at least the 1960s. The current study evaluated the training process as well as the use of the skills by course participants on a dedicated behaviour therapy course in Ireland. A cross sectional study was undertaken, using postal distribution of a questionnaire based on a similar study undertaken in the UK. The sample comprised all graduates of the course from 1986 to 1999 (N = 257). Thirty-three per cent of respondents report using their behaviour therapy skills to a high level since completion of the course. Approximately 53% of respondents reported having changed job since completion of the course. Significant differences emerged within this group in relation to their ratings of the importance of various elements of the course and also in relation to the perceived enhancement of both general clinical skills (P = 0.000) and specific behaviour therapy skills (P = 0.000) as well as the rating of current skills (P = 0.015) and the respondents' use of behaviour therapy since completing the course (P = 0.015). While a high proportion of participants had some prior theoretical or experiential exposure to behaviour therapy, the findings report high satisfaction levels with the training they received. However, the survey signals deficits in terms of the usage of the skills. While on the one hand, the general and specific skills acquired while on the course seem to transfer quite easily across functions and levels of responsibility, it is of concern that over a 13 year period, only 17% of respondents indicated that behaviour therapy was the main focus of their work. The implications of these findings are discussed in relation to policy and practice issues. PMID- 15788039 TI - Characteristics and staff resources of child and adolescent psychiatric hospital wards in Finland. AB - The aim of this study is to describe structural characteristics and staff resources of child psychiatric and adolescent psychiatric hospital wards in Finland. The target group of the survey consisted of 69 child and adolescent psychiatric hospital units in Finland. Information was obtained from 64 units (93%). Most of the wards were based on 24-h-a-day provision. There were only 7 day-treatment programmes including two family wards. When compared internationally, the numbers of units, beds and staff levels were high in Finland, with all members of staff qualified. The nurse-patient ratio and psychiatrist resources were rather satisfactory. However, in many units there was a lack of psychologists, social workers and occupational therapists. General recommendations and guidelines for staff resources in child and adolescent hospital treatment wards are warranted. PMID- 15788040 TI - Training in cognitive behavioural interventions on acute psychiatric inpatient wards. AB - There has been a drive towards addressing the types of care and therapeutic interventions available to people with serious mental illness, which is reflected in the latest government mental health policy initiatives. Recent evidence strongly supports the implementation of psychological and social interventions for people with psychosis, and in particular the use of cognitive behavioural techniques. Until now, the main focus has been on people living in the community. This study examines the delivery of psychosocial interventions training to qualified psychiatric nurses and unqualified staff on seven acute psychiatric admission wards in London, UK. The approach had the strength of on-site delivery, follow-up role modelling of the interventions and clinical supervision. Despite this, in some cases the training was less successful, mainly because of staffing and leadership weaknesses. The impact of training in these methods and the implications for mental health education and practice development are discussed. PMID- 15788041 TI - Evaluation of a postgraduate training programme for community mental health practitioners. AB - Government guidelines on mental health care in England have considerable implications for the level of competency required by the mental health workforce. Implementing these changes requires the widespread introduction of training initiatives whose effectiveness in improving staff performance need to be demonstrated through programme evaluation. This exploratory study evaluates the impact of a 2-year mental health training programme by measuring skill acquisition and skill application, by identifying the key ingredients for facilitating the transfer of learning into practice, and by examining differences in outcome between the academic and the non-academic students. High skill acquisition and application was reported in the majority of interventions, however, low skill application was reported for some key interventions (assertive outreach, dual diagnosis). Statistically significant differences were found between student cohorts in one intervention for skill acquisition (crisis intervention) and two interventions for skill application (client strengths model; medication management). The main ingredients for facilitating transfer were found to be the credibility of the trainers and training alongside colleagues from their own workplace. Some of the possible explanatory factors for these findings are discussed. PMID- 15788042 TI - Reasons for admission and their implications for the nature of acute inpatient psychiatric nursing. AB - The purpose of acute inpatient psychiatric care, and nurses' role within that, are in need of clarification and restatement in order to provide a framework for practice, education, research and development. Inpatient psychiatry has suffered from a paucity of research in recent years. In addition, being a complex system, involving multiple professions with differing ideologies, means that widely accepted succinct descriptions of its purpose are hard to achieve. Yet such a framework is essential to support positive attitudes to patients and for good staff-management relationships. Using an oblique strategy, this paper defines the function of acute inpatient psychiatry, and the role of psychiatric nurses, via a structured examination of the literature on reasons for admission to acute inpatient psychiatric wards. Seven such reasons were discovered and are described: dangerousness, assessment, medical treatment, severe mental disorder, self-care deficits, respite for carers, and respite for the patient. Acute inpatient psychiatric nurses are therefore: providing safety for the patient and others; collecting and communicating information about patients, giving and monitoring treatment; tolerating and managing disturbed behaviour; providing personal care; and managing an environment where patients can comfortably stay. The implications for psychiatric nursing are discussed. PMID- 15788043 TI - Antipsychotic depot medication and attitudes of community psychiatric nurses. AB - Utilization of long-acting antipsychotic injections (depots) shows wide regional variation. In many countries, community psychiatric nurses (CPNs) administer depots but their concerns and attitudes regarding these drugs are seldom considered. We aimed to investigate attitudes and knowledge towards depots in a cross-sectional survey of CPNs in London, and compare them with those of psychiatrists obtained in a previous study. Three subscales of a depot attitude/knowledge questionnaire were used with additional items which referred to aspects of the CPN role. Participants were 70 CPNs who attended an academic meeting. Most CPNs reported that they were involved in treatment decisions (78%) although some CPNs seldom asked their patients about side effects (19%) and felt that they did not have sufficient time for consultations (23%) or training (23%). Several CPNs believed that depots are old fashioned (34%) and stigmatizing (44%). Compared to psychiatrists, CPNs believed more that depots compromised patient autonomy (28%, P = 0.003) and were coercive (42%, P < 0.001). Familiarity with depots and their knowledge of side effects were positively associated with favourable attitudes. CPNs have several strongly endorsed attitudes towards depot medication. Interprofessional group differences also exist which may undermine the treatment process. Training/refresher courses about depots should highlight systematic treatment decision-making and side effect monitoring which, in turn, may improve professionals' attitudes, knowledge and clinical monitoring of depots. PMID- 15788044 TI - Exploring the concept of recovery in schizophrenia. AB - The concept of recovery in psychosis has gained much momentum in recent years in the UK. Current government policy describes its underpinning philosophy as the way forward for mental health services. Many mental health professionals now claim to embrace this concept yet fail to make the desired impact upon the care and treatment of individuals with schizophrenia. This article reviews some of the literature and explores what the concept of recovery means. The formal evidence will be augmented with personal accounts about recovery written by individuals who have schizophrenia. In doing so the main components that appear to have influenced the recovery process will be highlighted, and the implications for mental health nurses and practitioners will be discussed. PMID- 15788045 TI - Still invisible after all these years: mental health nursing on the margins. PMID- 15788046 TI - Predictors for successful angioplasty of chronic totally occluded coronary arteries. AB - AIMS: Reevaluation of clinical and angiographic predictors for percutaneous recanalization of coronary chronic total occlusion (CTO) using current techniques with conventional PTCA wires and balloons. METHODS AND RESULTS: We studied 253 consecutive patients with 283 lesions who underwent attempted PTCA of CTO (mean time since occlusion 33 months, range 3-150 month). Immediate procedural success rate was 84.8% (95% CI = 80.3%-88.6%). Multiple clinical and angiographic characteristics were evaluated as possible predictors of success/failure. Multiple logistic regression analysis revealed that a tapered morphology (P < 0.001, OR = 6.1; 95% CI = 2.1-18.2), 1 year) in consecutive, nonrandomized, patients who received intramyocardial VEGF-2. METHODS: Thirty patients with intractable CCS class III or IV angina and no options for revascularization underwent direct intramyocardial GT of VEGF-2 naked DNA via limited thoracotomy at total doses of 0.2, 0.8, or 2.0 mg. Patients were followed for clinical events after 1 year by hospital records, follow-up visits or telephone contact. Due to one perioperative death, 29 patients were followed. RESULTS: At a mean follow-up of 751 +/- 102.5 days (range 459-959) there were four deaths (13.8%), five myocardial infarctions (MIs) (17.2%), and seven revascularization procedures (24.1%). There were 15 hospitalizations in 12 patients. At the end of the follow-up period no patient (0%) had CCS class IV angina, 3 patients (11.5%) had class III angina, and 23 (88.5%) had class I to II angina. There were two new diagnoses of cancer. CONCLUSION: Transthoracic intramyocardial injection of VEGF-2 is associated with an improvement of symptoms of angina in the majority of patients beyond the first year of treatment. Major clinical events such as death, MI, and repeat revascularization are uncommon during the first year but more frequent after 1 year at a rate consistent with the severity of underlying disease in this population with advanced atherosclerosis. The majority of events were the result of progression of disease in areas of the heart remote from the site of GT. A large randomized trial is planned to determine the efficacy of intramyocardial VEGF-2 injections in inoperable patients. PMID- 15788052 TI - Predictors of bleeding complications and thrombocytopenia with the use of abciximab during percutaneous coronary intervention. AB - Abciximab (c7e3; ReoPro; Centocor, Inc., Pennsylvania, USA), a Fab fragment of the chimeric human-murine monoclonal antibody directed against platelet glycoprotein IIb/IIIa receptors, has been shown to reduce acute ischemic complications in patients undergoing percutaneous coronary interventions (PCI). However, its use has been associated with an increased incidence of bleeding events. The aim of this study is to examine the frequency of bleeding complications and to identify clinical variables associated with it in our patients. We prospectively recorded incidence of bleeding complications in 344 consecutive patients who underwent PCI with adjunctive use of abciximab (Ab). Clinical predictors for patients who developed bleeding complications with Ab were compared with the nonbleeders. Major bleeding occurred in six patients (1.7%) one of whom had pulmonary hemorrhage and one had intracranial hemorrhage. Minor bleeding occurred in 20 (5.8%) patients. Thrombocytopenia was seen in 13 (3.9%) patients of which four (1.2%) were mild, four (1.2%) were severe, and five (1.5%) were profound. Multivariate analyses showed that female gender and the "bail-out" use of Ab were risk factors for bleeding complications. The rate of major bleeding complications and thrombocytopenia were low and comparable with published trials. Female patients and those who received Ab as a "bail-out" were at higher risk of developing bleeding complications. PMID- 15788053 TI - Percutaneous angioplasty of the subclavian artery in patients with mammary coronary bypass grafts. AB - Severe coronary artery disease often coexists with peripheral vascular atherosclerosis. The assessment of the supra-aortic circulation in candidates for surgical coronary revascularization is of clinical relevance, and after intervention the patency of the supra-aortic trunks should be regularly evaluated in the clinical follow-up. We describe three cases of coronary artery disease treated with surgical revascularization using the internal mammary artery and concomitant atherosclerotic disease of the subclavian artery. The clinical presentation, the diagnostic procedures, and the therapeutic approach by means of percutaneous transluminal angioplasty of the subclavian artery are described and discussed. PMID- 15788054 TI - Percutaneous treatment of dissection of the ascending aorta occurring as a complication during coronary angioplasty of a saphenous vein bypass graft. AB - Aortocoronary dissection can occur as a complication of angioplasty of native coronary arteries. This case report is of aortic dissection occurring as a complication of percutaneous coronary intervention of proximal anastomoses of a saphenous vein bypass graft. The aortic dissection that had progressed retrogradely into the ascending aorta was treated percutaneously by stenting in the saphenous vein graft with a membrane-covered stent. PMID- 15788055 TI - The use of brachytherapy to treat renal artery in-stent restenosis. AB - The percutaneous treatment of renal artery stenosis has become the accepted revascularization strategy by most physicians treating this disorder. Unfortunately, as renal artery angioplasty and stent implantation become increasingly prevalent the Achilles heel of angioplasty, in-stent restenosis, also rises. There are currently no data suggestive of the optimal treatment strategy for renal artery in-stent restenosis. However, given the similarities in the pathophysiology between renal artery and coronary artery in-stent restenosis, brachytherapy is considered a reasonable option. This is the strategy that has been suggested and used by a number of operators. This case report describes two examples of renal artery in-stent restenosis treated with angioplasty and brachytherapy. PMID- 15788056 TI - Early initiation of statin therapy in acute coronary syndromes: a review of the evidence. AB - Acute coronary syndromes (ACS), such as myocardial infarction and unstable angina, are leading causes of death in developed countries. The risk of recurrent adverse events, rehospitalization, and death remain high in the weeks to months following ACS. Large secondary prevention trials have shown that the initiation of statin therapy within 3-6 months after hospitalization for ACS decreases the risk of recurrent cardiovascular events and death. Although the precise mechanisms behind these clinical benefits are unknown, data from human and animal studies have implicated statins in inflammatory response modulation, plaque stability, thrombus formation, and endothelial function. Several observational studies have demonstrated cardiovascular mortality and morbidity benefits in patients with ACS who were placed on statins within hours to days of their event. Three recent prospective controlled trials confirmed these benefits and demonstrated that moderate doses of statins are safe when used in patients with ACS. We recommend the initiation of statin therapy in all ACS patients prior to hospital discharge. PMID- 15788057 TI - Sirolimus-eluting stent implantation for chronic total occlusion of the left main coronary artery. AB - Chronic total occlusion of the left main coronary artery (LMCA) is rare. Recently, percutaneous coronary intervention has been increasingly applied to unprotected LMCA lesions. We describe a patient with chronic total occlusion of the LMCA who was successfully treated with bifurcation stenting with sirolimus eluting stents. PMID- 15788060 TI - Evaluating a family-centred intervention for infant sleep problems. AB - AIM: This paper reports a study to describe changes in parents' distress after a family-centred intervention for sleep problems of infants. BACKGROUND: Infant sleep problems are common and are related to depressive symptoms in mothers, but their impact on fathers has rarely been studied. Because childhood sleep problems and parental distress are associated, their interdependence should be recognized in research and in paediatric sleep practice. METHODS: All children hospitalized for sleep problems in a hospital in Iceland in 1997-1998 and their parents were studied using a pre- and post-test quasi-experimental design. The sample consisted of 33 infants (6-23 months of age), 33 mothers and 30 fathers. Parents' distress was assessed before and after treatment with regard to: (1) fatigue and resulting symptom distress; (2) parenting stress; (3) state-anxiety; and (4) depressive symptoms. Infants were treated for a variety of sleep problems by a paediatric nurse. The parents were simultaneously treated for distress by either the paediatric nurse or a specialist, depending on the nature of their problems. RESULTS: Mothers and fathers experienced a high degree of distress before the intervention, with no significant difference between them. Two months after the intervention both parents' distress had significantly improved. Parents' degree of distress was at a psychopathological level before the intervention but was reduced to population norms 2 months after the intervention. The paediatric nurse intervention was sufficient to reduce distress for 83% of parents. CONCLUSIONS: Health care professionals who care for infants with sleep problems should pay attention to the distressed responses of parents and support their recovery. An intervention such as that described here could be used by nurses for this purpose. PMID- 15788061 TI - Countertransference and self-injury: a cognitive behavioural cycle. AB - AIM: This paper discusses the emotional, cognitive and behavioural effects of self-injury on nurses as helpers, and shows the usefulness of a cycle that can affect care provision for this group of people. BACKGROUND: People self-injure for many different reasons, such as feeling angry, sad, guilty or frightened, and these emotions are often linked to feeling helpless, powerless or out of control. Self-injury has often been reported as a coping strategy to gain control. Psychoanalytic and cognitive behavioural concepts have been used to understand why people self-injure and also inform intervention strategies. Unfortunately, negative emotional responses in professionals may interfere with the effectiveness of any therapeutic relationship. DISCUSSION: Negative emotional responses from nurses can affect the way they think about and behave towards clients who self-injure. During clinical supervision or education, nurses' thoughts can be challenged to become less negative, so that their resulting behaviour can also become less punitive. Non-punitive or more positive behaviour can in turn challenge some of the negative self-beliefs of clients. CONCLUSIONS: Knowledge about countertransference when working with people who self-injure may reduce nurses' negative thoughts and behaviours, which may result in improved client care. PMID- 15788062 TI - Read, Think, Do!: a method for fitting research evidence into practice. AB - AIM: This paper discusses a process for research utilization that overcomes well known barriers in order to influence clinical decision-making and practice change. Read, Think, Do! is a problem-solving approach to research utilization and practice development which has the potential to overcome barriers to research utilization. BACKGROUND: Any process for research utilization at the practice level needs to overcome numerous barriers in order to influence clinical decision making and practice change. Access to research-based knowledge is an obvious first step in the evidence-based approach to care delivery, but is clearly inadequate alone in influencing the improvement of practice. DISCUSSION: Read, Think, Do! acknowledges the complexity of problem-solving processes from the outset by looking for (1) the evidence, (2) assessing the value to practice, and (3) addressing the social and cultural milieu of the practice setting to ascertain the best strategies for initiating and sustaining practice change. This approach draws distal forms of empirical knowledge that have the capacity to improve patient outcomes into the proximal knowledge base of the clinical nurse. This is achieved by collaboration, planning and evaluation involving all levels of staff and a specialist facilitator, the Clinical Nurse Consultant in evidence based practice. CONCLUSION: Read, Think, Do! is a method of research utilization and practice development that has the potential to overcome barriers to research utilization and avoid the "misplaced concreteness" that can occur when trying to fit empiricism into practice. By addressing the breadth and diversity of issues surrounding research utilization in a systematic manner it presents a sustainable method for practice change informed by evidence. PMID- 15788063 TI - Understanding the workplace culture of a special care nursery. AB - AIM: This paper presents findings from the first phase of a research study focusing on implementation and evaluation of emancipatory practice development strategies. BACKGROUND: Understanding the culture of practice is essential to undertaking effective developments in practice. Culture is a dominant feature of discussions about modernizing health care, yet few studies have been undertaken that systematically evaluate the development of effective practice cultures. METHOD: The study intervention is that of emancipatory practice development with an integrated evaluation approach based on Realistic Evaluation. The aim of Realistic Evaluation is to evaluate relationships between Context (setting), Mechanism (process characteristics) and Outcome (arising from the context mechanism configuration). This first phase of the study focuses on uncovering the context (in particular the culture) of the Special Care Nursery in order to evaluate the emancipatory practice development processes and outcomes. Data collection methods included survey, participant observation and interview. Cognitive mapping, constant comparative method and coding were used to analyse the data. Findings. Four key categories were identified: Teamwork, Learning in Practice, Inevitability of Change and Family-Centred Care and collectively these formed a central category of Core Values and Beliefs. A number of themes were identified in each category, and reflected tensions that existed between differing values and beliefs within the culture of the unit. CONCLUSION: Understanding values and beliefs is an important part of understanding a workplace culture. Whilst survey methods are capable of outlining espoused workplace characteristics, observation of staff interactions and perceptions gives an understanding of culture as a living entity manifested through interpersonal relationships. Attempts at changing workplace cultures should start from the clarification of values held among staff in that culture. PMID- 15788064 TI - Effects of an adaptation training programme for patients with end-stage renal disease. AB - AIMS: This paper reports a study investigating the effectiveness of an adaptation training programme (ATP) to help patients with end-stage renal disease (ESRD) to cope with illness-related stresses and, thus, to alleviate depression and improve quality of life. BACKGROUND: Patients with ESRD who receive dialysis must confront the burdens of long-term illness and numerous treatment-associated stressors. The ability of these patients to cope with and adapt to these stresses, whether related to their medical regimen or to the demands of daily life, has an important influence on physical and psychological well-being. METHODS: The study was a randomized controlled trial using a convenience sample of 57 eligible, fully informed and consenting patients with ESRD who were assigned to experimental (ATP plus usual care) or control (usual care) groups. Participants in the ATP took part in weekly small group sessions over an 8-week period and monthly follow-up to help them to cope with stressors. A clinical nurse specialist and an experienced psychotherapist led them in three small groups (8-10 per group). Participants in the usual care group received routine care. Instruments comprised the Haemodialysis Stressor Scale, Beck Depression Inventory and Medical Outcomes Study SF-36. Data were collected at baseline and at three months following the intervention. RESULTS: The major stressors for these patients were limitations on time and place related to employment, limitations on fluid intake, transport difficulties, loss of bodily function, length of dialysis treatment, and limitation of physical activities. The ATP had a beneficial effect on perceived stress (P = 0.005), depression (P = 0.001) and quality of life (P = 0.02) 3 months after the intervention. CONCLUSIONS: This study supports the effectiveness of an ATP to decrease stress and depression levels, and to improve the quality of life of ATP patients receiving haemodialysis. PMID- 15788065 TI - Beliefs about health and diabetes in men of different ethnic origin. AB - AIM: This paper reports the findings of a study exploring the health and illness beliefs of men with diabetes, who were from different cultural backgrounds and living in Sweden. BACKGROUND: No studies have been reported that have focused on the beliefs about health and illness in men with diabetes mellitus of different ethnic origin. Beliefs may affect self-care and care-seeking behaviour. METHOD: An explorative study design and purposive sampling procedure was used. Focus group interviews were held with 35 men with diabetes and aged between 39 and 78 years. Fourteen participants were born in Arabic countries, 10 in former Yugoslavia and 11 in Sweden. FINDINGS: Important factors for health were the ability to be occupied/employed and economically independent and, especially among Arabs and former Yugoslavians, sexual functioning. Swedes focused on heredity, lifestyle and management of diabetes, while non-Swedes claimed the influence of supernatural factors and emotional stress related to the role of being an immigrant and migratory experiences as factors related to development of diabetes and having a negative influence on health. Swedes and Arabs described health as "freedom from disease" in contrast to many former Yugoslavians who described health as "wealth and the most important thing in life". Knowledge about diabetes was limited among the men studied, but Arabs showed an active information-seeking behaviour compared with Swedes and former Yugoslavians. Non Swedish respondents, particularly Arabs, had sought help from health care professionals to a greater extent than Swedes, who were more likely to use self care measures. CONCLUSION: Being occupied/employed and having knowledge about the body and management of diabetes are important for positive health development. There are dissimilarities in beliefs about health and diabetes that influence self-care behaviour and health care seeking. Men's cultural backgrounds and spiritual beliefs need to be considered in diabetes care. PMID- 15788066 TI - Maternal-fetal attachment: an integrative review. AB - AIMS: This paper presents an integrative review of the literature that aimed to (a) review, categorize and critique the published research on maternal-fetal attachment (MFA), (b) analyse and interpret the MFA findings and (c) evaluate the state of knowledge on MFA and provide directions for future research. BACKGROUND: Previous researchers have examined MFA in independent studies using a variety of research designs. There was a need to synthesize the findings across studies in order to evaluate the knowledge developed about MFA since the construct was created. METHODS: Five stages for conducting an integrative review were carried out. The research problem was identified; data were collected; data were evaluated; findings were analysed and interpreted. Forty-one studies of MFA were located using the online databases of Medline, International Nursing Index, Cumulative Index of Nursing and Allied Health Literature, and Psychological Information. The keywords for the search were MFA, parental-fetal attachment, and prenatal attachment. FINDINGS: Of the 41 studies reviewed, there were 152 findings relevant to the analysis. Of the 152 findings, 70 were statistically significant, while 82 were not. CONCLUSIONS: Research focusing on MFA has begun to create a body of scientific knowledge. This needs to be expanded through continued research using a variety of designs. PMID- 15788067 TI - Nutritional screening and assessment tools for use by nurses: literature review. AB - AIM: This paper reports a literature review describing the range of published tools available for use by nurses to screen or assess nutritional status of patients/clients, and to examine whether the validity, reliability, sensitivity, specificity and acceptability of the tool have been investigated. Tools developed specifically for use with older adults are not considered in this review. A screening/assessment tool is described as a tool that uses a questionnaire-type format, contains more than one risk factor for malnutrition and gives an assessment of risk. BACKGROUND: The incidence of malnutrition in people cared for by nurses is high and screening or assessment tools are often used to identify those with, or at risk of, malnutrition. METHODS: A comprehensive literature review methodology was employed. A range of electronic databases was searched from 1982 to 2002. Search terms incorporating "nutrition", "screening", assessment, feeding, instrument, tool, validity, reliability, sensitivity and specificity were combined. Manual searches were also conducted. RESULTS: Seventy one nutritional screening/assessment tools were identified of which 35 were reviewed. Tools not reviewed included those which incorporated significant use of biochemical measures (8), included complex anthropometric measures (3), were concerned specifically with ingestion (4) or were designated for use with an older population (21). The tools reviewed use a wide variety of risk factors for malnutrition, varying from anthropometric measurements to socio-cultural aspects of eating behaviour. Some identify an action plan based on the score obtained. Many have not been subjected to validity and/or reliability testing and yet appear to be in use in clinical practice. The sensitivity, specificity and acceptability of the tools are often not investigated. CONCLUSION: There are many published nutritional screening/assessment tools available for use by nurses to screen or assess the nutritional status of patients/clients. Many have not been subject to rigorous testing. Future work should consider a more standardized approach to the use of these tools. PMID- 15788068 TI - The Nurse Educator's clinical role. AB - AIM: This paper reports a two-phase descriptive study exploring the clinical role of the nurse educator in Malta. BACKGROUND: Previous studies indicate a number of similarities and differences in the clinical role of nurse educators by country of practice. These include importance assigned to the role, factors inhibiting/facilitating the role, means to eliminate barriers to the role, and perceptions of the ideal role. DESIGN AND METHODS: Data were collected using both quantitative and qualitative strategies. The quantitative phase involved asking all educators to fill in a time log of their academic and clinical activities for a 2-week period. In the qualitative phase, the first author interviewed five educators, five nurses and five students about their perceptions of factors which impact the nurse educator's clinical role, as well as what the ideal clinical role of the nurse educator should be. FINDINGS: Maltese nurse educators allot minimal time to their clinical role. Main reasons cited included workload, perceived lack of control over the clinical area, and diminished clinical competence. Nurse educators who frequented the clinical settings (who were either university or joint university and health service employees) where the study took place perceived that employment inequities among the various categories of nurse educators played an important role in the amount of time dedicated by each group to their clinical roles, and the importance individuals in these groups assigned to that role. The majority of interviewees saw the current role of nurse educators in Malta as preparing students for successful completion of the didactic sections of their programme, rather than preparing them with all the knowledge and clinical skills necessary to be competent practitioners. Participants considered that, when in clinical areas, nurse educators did focus on their students, as they should. However, they also thought that they often did not take the opportunity to forge links with professional staff. CONCLUSION: The clinical role of the Maltese nurse educator needs to be more multifaceted in approach. PMID- 15788069 TI - Programme-related stressors among part-time undergraduate nursing students. AB - AIM: The aim of this paper is to report a study exploring the perceived stressors identified by a group of 70 students who undertook a part-time degree at one Irish university. BACKGROUND: In the literature on stress, part-time nursing students who are undertaking continuing education programmes appear to have received little attention. Stress amongst nurses is evident within the nursing literature but little information is available on the specific stressors that affect Registered Nurses who undertake further academic study. Anecdotally, students attending part-time programmes while working full-time report high levels of stress. METHOD: Quantitative methods were used. While many instruments exist to measure overall stress, this study aimed to explore student's perceptions of specific stressors associated with academic study. We used a questionnaire developed from the literature on the topic. RESULTS: Factors related to writing assignments at degree level, fulfilling personal needs and academic demands, were perceived as major stressors by these students. Factors of little concern were financial issues and attendance on the programme. Individual items receiving highest mean scores were: trying to balance work commitments and the required study (mean 3.89, sd = 1) and the prospect of the final examination (mean 3.86, sd = 1). This study was limited by the use of convenience sampling and self-report methods. Larger studies are required to support the findings. In addition, student stress was not observed or measured. CONCLUSION: Those involved in the delivery of nurse education programmes to part-time students need to consider the impact of the workload on student welfare, and to prepare students for demands of the programme. PMID- 15788070 TI - SARS: coping with the impact at a community hospital. AB - AIM: This paper presents the findings of a staff survey conducted at a 350-bed acute care facility located on the periphery of Toronto, Canada. BACKGROUND: Toronto's severe acute respiratory syndrome (SARS) crisis resulted in trauma-like effects at hospitals hardest hit by the disease. A systematic examination of the impact on staff working in hospitals that saw relatively few cases, while maintaining the precautions associated with elevated alert levels, has not been undertaken. METHODS: A questionnaire was distributed for 1 month commencing 17 April 2003 and 300 completed responses were obtained (approximately one in six staff members). The data collected included demographic and occupational information, in addition to perceptions of SARS' impact on patient care, factors contributing to adverse impacts on patient care, working conditions, decision making, communication and relations, sources of support, and the impact on workers' lives outside work. Items for these sections were developed by a multi disciplinary team of health care workers and hospital administrators. RESULTS: In the absence of pre-SARS normative data for the survey, demographic and occupational variables were used to look for patterns of differences between relevant subgroups of respondents. Statistically significant differences were found for gender (73.9% women), nurses (24.7%) vs. others, doctors (20.3%) vs. others, older (40 years or older, 60.0%) vs. younger persons, emergency or intensive care unit workers (8.0%) vs. others, and those employed fewer years at the hospital (less than 5 years: 46.2%) vs. five or more years. These differences varied across the following domains: factors adversely affecting patients, communications, support, working conditions, decision-making and, to a lesser extent, impact on life outside work. While all groups found SARS stressful, nurses reported a greater impact on morale and job satisfaction. Nurses relied more on peer support than doctors, felt less informed and less involved in decision-making than doctors felt, and were more likely to report that infection control procedures were not strict enough. CONCLUSIONS: The between-group differences and the pattern of these differences clearly illustrate the polarizing and stressful impact SARS had at a hospital with only a small number of probable or suspect cases. The clear differences between groups defined by demographics, professions and clinical roles suggest a subtle and pervasive secondary impact of the SARS outbreak, with repercussions health care facilities must contend with while maintaining increased levels of vigilance in the wake of SARS. PMID- 15788071 TI - A response to D. Von Ah, S. Ebert, A. Ngamvitroj, N. Park & D.-H. Kang Predictors of health behaviours in college students. Journal of Advanced Nursing 48(5), 463 474. PMID- 15788076 TI - Progressing evidence-based practice: an effective nursing model? AB - AIMS: This paper presents findings from telephone interviews completed with link nurses 2 years into the project to explore how participation progressed achievement of evidence-based practice where the link nurses worked. BACKGROUND: In 2001, an innovative practice development initiative was launched in Scotland. A national network of experienced nurses from across the country was recruited to form the inaugural Community of Practice. This involved describing gerontological nursing, pioneering a nurse-sensitive methodology to craft care guidance that reflects the agreed practice model, and constructing a virtual college based on a situated learning model. METHODS: A volunteer sample of link nurses took part in telephone interviews exploring experiences of using the virtual college and the extent to which the description of gerontological nursing and the first best practice statement on nutrition had influenced practice. FINDINGS: Five components (themes) were identified as facilitating the attainment of evidence based practice. These focussed on confidence-building and the positive benefits of achieving vision and clarity for gerontological nursing. Membership of a national Community of Practice afforded status and strengthened sense of professional identity. The inclusive knowledge synthesis methodology used to prepare, pilot and support implementation of the best practice statement was highly valued. Progress towards evidence-based practice in all affiliated areas was reported. Major challenges for nurses in participating in the virtual college included the absence of a learning-at-work culture, lack of time and doubts about the legitimacy of internet-based learning. CONCLUSION: The evaluation indicates the potential merits of e-practice development, particularly for nurses who feel geographically and professionally isolated or disenchanted with available continuing professional development opportunities. Participation in the virtual college appeared to enrich practice and foster a culture of change. PMID- 15788077 TI - Identifying HIV/AIDS primary care development needs. AB - AIM: This paper reports a study aimed at identifying the primary health care experiences of people living with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) in Malaysia. The rationale behind the study was to enable informed action for developing more responsive and effective primary care. BACKGROUND: Reports such as from the World Health Organisation forecast sharp escalations in the incidence of HIV/AIDS in Malaysia and the Asia-Pacific region within the next few years. With sparse information on the course of infection on the local population and an understanding of health care needs of those afflicted, health services would be ill-prepared for projected increases. METHOD: Semi-structured interviews were conducted with a convenience sample of 99 patients attending two major HIV/AIDS clinics in Malaysia. FINDINGS: Several gaps in care provision were highlighted, such as with treatment/consultation facilities and availability and accessibility of information. What is also evident is that there are a number of good support services available but not well publicized to those in need of them. That includes health professionals who could be making appropriate referrals. The lack of communications and inter professional working appears to be part of the problem. CONCLUSION: The findings provide baseline data and preliminary insights to government and other service providers towards advancing, optimizing and refining existing policies and infrastructure. Although the availability of a number of primary care facilities have been identified, the study indicates the need for more effective co ordinated efforts with clear leadership to pull together scarce resources towards the aim of some degree of seamless primary care provision. It is suggested that nurses would be well placed for such a role in view of the nature of their education and training that helps prepare them for the multi-faceted role. PMID- 15788078 TI - Patient participation on a ward for frail older people. AB - AIMS: The paper reports a study whose aims were to explore the meaning of participation for older people in hospital and their health care workers and ways in which staff can enhance patient participation in their care. BACKGROUND: Participation in care is a concept that is central to nursing practice. However, little is known about patient and staff views of participation and how it takes place in practice. METHODS: Action research was used, and data were collected by means of focus groups and in-depth interviews. Three focus groups and 24 interviews were undertaken with staff, and 19 interviews were undertaken with patients. The main action undertaken with staff aimed at raising their awareness of participation. In addition small changes in practice were introduced, such as the use of a patient biographical booklet. FINDINGS: The findings suggest that participation is a dynamic process that is integral to the work of nurses and carers. The process is enacted through the themes of facilitation, partnership, understanding the person, and emotional work. CONCLUSION: The study demonstrates how nurses can become increasingly aware of the potential for facilitation and creation of opportunities for participation. It is crucial that practitioners working with older people develop a deeper understanding of participation and are also empowered to act in ways that fit with the needs of this specific client group. PMID- 15788079 TI - Caring for a child with cancer: a longitudinal perspective. AB - AIMS: This paper reports a study to evaluate caregiving demands among mothers and fathers of children with cancer, parents' well-being, and how the parents perceive the health status of their child over a period of 18 months. BACKGROUND: Cancer among children has psychological impact on all family members. Most previous research has focused on distress, coping patterns and social support, and less is known about how parents are dealing with the illness experience over time. No study was found that evaluated caregiving on a daily basis, or the impact of the cancer on wellbeing and perceptions of health within families. METHODS: Data were collected between 1999 and 2001. Parents of 26 Icelandic children under 18 years with cancer completed questionnaires about caregiving activities, general wellbeing and health perceptions. Descriptive statistics and Repeated Analyses of Variance were conducted at baseline, 12 and 18 months. RESULTS: The most time-consuming and difficult caregiving activities for both mothers and fathers were giving emotional support to the child with cancer, and to other children in the family. Mothers also found it difficult and time consuming to manage behavioural problems and to structure and plan activities for the family. Fathers found it difficult to manage work and organize care for the child at the same time, and to give their partner emotional support. When evaluated over time, mothers' caregiving demands differed statistically significantly from fathers', and fathers' caregiving demands and health perceptions changed statistically significantly over the 18 months of the study. CONCLUSION: These findings underline the long-term impact of children's cancer on their families. Interventions focusing on emotional support, parents' wellbeing, and how families perceive the health of their child with cancer might benefit the family as a whole. PMID- 15788080 TI - Cancer patients' descriptions of their nursing care. AB - AIM: This paper reports a study of cancer patients' descriptions of nurses and nursing care. BACKGROUND: Nurses lament their poor representation in the media, and campaigns to improve their portrayal have been initiated. Media portrayal of nurses might be more realistic if patients' descriptions of nursing care were incorporated. METHOD: Qualitative data from an instrument development study were analysed. A total of 461 patients answered the question, 'In general, how do you feel about nurses?' The data were analysed by the constant comparative method and grounded theory coding techniques. FINDINGS: A typology of four concepts reflecting cancer patients' descriptions of their nursing care emerged from the data. The concepts were: laudable, caring, professional and outcomes. The concept laudable refers to commendable qualities of the nurse and nursing care. Caring refers to the nurse showing compassion, concern and kindness. Professional refers to the nurse as meeting expected standards of knowledge, skill and demeanour. Outcomes refer to the affective, cognitive, or physical effects attributed to nursing care. Both positive and negative instances of the concepts were included in the analysis. Examples of each concept using patients' own words are given. CONCLUSIONS: These cancer patients held nurses in relatively high esteem. These findings could be disseminated to the public press as an example of what patients' value about nurses and nursing care. They also could be used in media efforts to recruit and retain nurses. PMID- 15788081 TI - Factors affecting the use of nursing information systems in Taiwan. AB - AIM: This paper reports a study to test a proposed model of factors (demographic variables, computer experience and perceived technology characteristics) influencing the efficient use of a computerized nursing care planning system by hospital clinical nurses. BACKGROUND: Although nursing information systems have been used in health care institutions, little research has explored the efficiency of computer use in nurses' daily practice. METHOD: A secondary data analysis was applied to focus on clinical nurses' daily computer use. The original data were collected from a medical centre in Taiwan to establish the reliability and validity of a scale to evaluate computerized nursing care plan systems. RESULTS: Younger nurses with more education, less computer knowledge and less usage pressure, and who perceived that they had less usage benefit, more education training and usability regarding system use, spent less time on the computerized nursing care plan. CONCLUSION: The results can be used for further organization of education programmes and as a reference for training strategies. Further examination and analysis of item content are suggested. PMID- 15788082 TI - Bourdieu's theory of practice and its potential in nursing research. AB - AIMS: This paper seeks to consider the utility of Bourdieu's "Theory of Practice" in nursing, and considers specifically its use as a framework for research exploring nurses' conceptualizations of illness and the patients in their care. Bourdieu's work uses the concepts of field, capital and habitus to explain interactions within the social world. This paper describes these concepts and their relationship with nursing is discussed using dementia care as an example. BACKGROUND: The work of French scholar Pierre Bourdieu has contributed to debates throughout the social sciences, but has had relatively little attention in the nursing literature. Pierre Bourdieu's work developed against a backdrop of change in the academic world. The emergence of the social sciences and the debate around objective and subjective styles of research were influential in the development of his "Theory of Practice". DISCUSSION: The importance of the conceptualization process is discussed, and the considerable potential influence of conceptualization on patient care is highlighted. Reflexivity is a cornerstone of Bourdieu's work, and is an important feature of nursing research. Examples of health care research using his work as a framework are discussed, and some of the challenges of the approach are outlined. CONCLUSIONS: The use of Bourdieu's "Theory of Practice" as a research framework could allow nurse researchers to explore the interactions of nurses with the structures, agents and symbols of illness within the field of care. This work could enhance understanding of how nurses view and react to patients in their care, and promote the development of practice innovations and policy change. The theory may, therefore, have much to offer future nursing research. PMID- 15788083 TI - Technology dependence and health-related quality of life: a model. AB - AIM: This paper presents a new theoretical model to explain people's diverse responses to therapeutic health technology by characterizing the relationship between technology dependence and health-related quality of life (HRQL). INTRODUCTION: Technology dependence has been defined as reliance on a variety of devices, drugs and procedures to alleviate or remedy acute or chronic health problems. Health professionals must ensure that these technologies result in positive outcomes for those who must rely on them, while minimizing the potential for unintended consequences. Little research exists to inform health professionals about how dependency on therapeutic technology may affect patient reported outcomes such as HRQL. Organizing frameworks to focus such research are also limited. MODEL: Generated from the synthesis of three theoretical frameworks and empirical research, the model proposes that attitudes towards technology dependence affect HRQL through a person's illness representations or commonsense beliefs about their illness. Symptom distress, illness history, age and gender also influence the technology dependence and HRQL relationship. Five concepts form the major components of the model: a) attitudes towards technology dependence, b) illness representation, c) symptom distress, d) HRQL and e) illness history. CONCLUSION: The model is proposed as a guide for clinical nursing research into the impact of a wide variety of therapeutic health care interventions on HRQL. Empirical validation of the model is needed to test its generality. PMID- 15788084 TI - Practical inquiry/theory in nursing. AB - AIM: This paper explores a social constructionist, pragmatist approach to inquiry and theory-building with a view to exploring its relevance for nursing as a practical discipline. BACKGROUND: Positivist and postpositivist inquiry approaches in practical disciplines have produced "detached" theories that lack relevance for everyday practice and so sustain the theory-practice gap. Both meta and mid-range theories tend to see practice as fixed or fixable rather than being enacted in a state of flux. DISCUSSION: Practical inquiry and theory are described structurally and as co-dependent processes. The research process is sensitive to the influence of context and consists of construction rather than capture. Practical theory is judged in terms of whether it helps people to "go on with" their lives. Practical inquiry/practical theory is superimposed on a previous nursing study in the field of mental health to illustrate how it can account for the processes of clinical research. In particular, the illustration demonstrates the surrender of researcher objectivity in the interests of collaborative understanding that occurs with practical inquiry/theory. Shared meaning arises as rich constructs of the research situation are developed that point to future possibilities for action for all those engaged in the research process. CONCLUSION: Practical inquiry/theory offers the means to conduct cogent, collaborative, developmental research, although further "trying out" is required. PMID- 15788085 TI - Meta-synthesis method for qualitative research: a literature review. AB - AIM: This paper discusses the purpose and stages of meta-synthesis and the epistemological status of knowledge generated from the technique. Particular attention is paid to exploring the contested areas of the method that remain. BACKGROUND: There is a growing interest in meta-synthesis as a technique for generating new insights and understanding from qualitative health care research. An increasing number of meta-synthesis papers are appearing in the nursing and midwifery literature. METHODS: Literature on the technique of meta-synthesis and examples of meta-synthesis papers were searched and reviewed. A meta-synthesis exercise was undertaken, and this informed reflection and critique of the method. FINDINGS: Meta-synthesis attempts to integrate results from a number of different but inter-related qualitative studies. The technique has an interpretive, rather than aggregating, intent, in contrast to meta-analysis of quantitative studies. Examples from the literature indicate that some aspects of the technique are not yet fully established. CONCLUSION: Despite the contingent nature of evidence gleaned from meta-synthesis and current lack of consensus about some of its aspects, meta-synthesis is an important technique for qualitative researchers and can deepen understanding of the contextual dimensions of health care. PMID- 15788086 TI - Preoperative education for orthopaedic patients: systematic review. AB - AIMS: This paper presents a systematic review whose aim was to describe the scope and methods of the current literature on preoperative patient education and to identify the effects of this education. BACKGROUND: Preoperative patient education is a common and important intervention in surgical nursing, yet there is very limited systematic evidence on its precise role. METHODS: The Medline, CINAHL, Eric, Psycinfo and Social Sciences Index databases and the Cochrane Library were searched, covering the period from the beginning of each database to April 2003. Studies were included if they concerned adult orthopaedic patients, preoperative nursing patient education and were based on randomized controlled or clinical trials. Meta-analysis was carried out where appropriate. RESULTS: We identified 11 articles involving 1044 participants. Most studies included one experimental and one control group; only two had more than one experimental and control group. The educational interventions varied widely, but the majority were based on written materials alone, or written materials in combination with other teaching methods. The most common outcome measures related to pain, knowledge, anxiety, exercises and length of stay, and the least common to self-efficacy and empowerment. The methodological quality of the studies varied. Almost all reported one or more statistically significant effects. Based on the findings of the meta-analysis, preoperative education appears to have some impacts on patients' anxiety and knowledge levels. CONCLUSIONS: The review clearly highlights the need for well-designed, methodologically sound research into the outcomes of patient education. It also points to the need to study patient education from the point of view of empowerment. PMID- 15788089 TI - Systems for grading the quality of evidence and the strength of recommendations II: pilot study of a new system. AB - BACKGROUND: Systems that are used by different organisations to grade the quality of evidence and the strength of recommendations vary. They have different strengths and weaknesses. The GRADE Working Group has developed an approach that addresses key shortcomings in these systems. The aim of this study was to pilot test and further develop the GRADE approach to grading evidence and recommendations. METHODS: A GRADE evidence profile consists of two tables: a quality assessment and a summary of findings. Twelve evidence profiles were used in this pilot study. Each evidence profile was made based on information available in a systematic review. Seventeen people were given instructions and independently graded the level of evidence and strength of recommendation for each of the 12 evidence profiles. For each example judgements were collected, summarised and discussed in the group with the aim of improving the proposed grading system. Kappas were calculated as a measure of chance-corrected agreement for the quality of evidence for each outcome for each of the twelve evidence profiles. The seventeen judges were also asked about the ease of understanding and the sensibility of the approach. All of the judgements were recorded and disagreements discussed. RESULTS: There was a varied amount of agreement on the quality of evidence for the outcomes relating to each of the twelve questions (kappa coefficients for agreement beyond chance ranged from 0 to 0.82). However, there was fair agreement about the relative importance of each outcome. There was poor agreement about the balance of benefits and harms and recommendations. Most of the disagreements were easily resolved through discussion. In general we found the GRADE approach to be clear, understandable and sensible. Some modifications were made in the approach and it was agreed that more information was needed in the evidence profiles. CONCLUSION: Judgements about evidence and recommendations are complex. Some subjectivity, especially regarding recommendations, is unavoidable. We believe our system for guiding these complex judgements appropriately balances the need for simplicity with the need for full and transparent consideration of all important issues. PMID- 15788090 TI - Policymaking based on CERs: changes in costs are not the same as changes in benefits. AB - BACKGROUND: Earlier cost-effectiveness studies showed the cost-effectiveness ratios for pneumococcal vaccination in preventing cases of Bacteremic Pneumococcal Disease (BPD) alone to vary between 11,000 and 33,000 per quality adjusted life year. If vaccination is also assumed to be effective in preventing cases of Non Bacteremic Pneumococcal Disease (NBPD) (at the same level of effectiveness), vaccinating all elderly persons becomes highly cost-effective or even cost saving. METHODS: The present article examines the effect of partial preventive power of the vaccine against cases of NBPD additional to its preventive power against cases of BPD, and the consequences this has in terms of cost-effectiveness. RESULTS: The analysis shows that even a fairly small additional preventive power against cases of NBPD leads to a dramatic and unexpected decrease in the cost-effectiveness ratio. CONCLUSION: Because a Cost Effectiveness Ratio (CER) is a ratio, changes in costs and changes in effects have rather different influences on its value. There is a linear relation between a change in costs and a change in CER if the effects are kept constant. This linear relation is not found on the effect side. Assuming that costs are constant, a change in effect will be different for low levels of effect than for high levels. PMID- 15788091 TI - Low dose docosahexaenoic acid protects normal colonic epithelial cells from araC toxicity. AB - BACKGROUND: The nucleoside analogue arabinosylcytosine (araC) has been used for many years in the treatment of acute leukemia. Evidence in the literature suggests that araC may inhibit the growth of human colon carcinoma cell lines as well. Because araC action interferes with normal nucleoside metabolism, it is highly toxic to a number of normal cell types including bone marrow and intestinal mucosa cells. Here we investigate whether the omega-3 fatty acid docosahexaenoic acid (DHA) could selectively target araC toxicity toward colonic tumor cells while protecting the normal cells in vitro. RESULTS: Cultures of normal rat colonic epithelial cells (4D/WT) and those transformed by v-src (D/v src) were supplemented with graded concentrations of DHA or arachidonic acid (AA) alone or in combination with araC. AraC was only 1.6 fold more toxic to D/v-src than 4D/WT in cultures without added fatty acids. Supplementing with as little as 3 muM of either AA or DHA increased araC toxicity by more than 30-fold in the tumorigenic cells. The toxic effect of araC on the normal cells was also increased by the fatty acid supplementation. IC50 values were decreased 1.7 fold by DHA in the 4D/WT cells but a more than 7-fold decrease was observed during AA supplementation. As a result, the therapeutic index of araC (IC50 normal/IC50 tumor) was more than 3-fold higher in the DHA than the AA supplemented cells. The expression of protein kinase C isoform epsilon was decreased in AA alone supplemented D/v-src cultures but in combination with araC decreased only in DHA supplemented 4D/WT cells. CONCLUSION: Low dose DHA supplementation may enhance araC chemotherapy in colon cancer while protecting normal tissues, possibly through control of PKC signalling pathways. PMID- 15788093 TI - What's in a virus? Folk understandings of hepatitis C infection and infectiousness among injecting drug users in Kings Cross, Sydney. AB - BACKGROUND: To explore folk understandings of blood borne virus infection and infectiousness among injecting drug users in Kings Cross, Sydney. METHODS: Observational fieldwork was conducted in Kings Cross over a four month period. In depth interviews with 24 current injectors and 4 key informants recruited from King Cross were undertaken. RESULTS: Hepatitis C (HCV) generated different meanings from HIV. HIV was considered "the dreaded" and generated fear of infection and dire disease progression. Whereas HCV was considered non-desirable but less threatening than HIV. The risks of transmitting HCV through sharing injecting paraphernalia was poorly understood. Some believed HCV infection was linked to poor hygiene and dirty water. Jaundice was mistakenly thought to indicate HCV infection and was used to gauge infectiousness. Many were confused about their current hepatitis C serostatus. Some participants thought they had a "dormant antibody" or that they had a "mild case" of infection. Participants were unsure what this meant for their own health or for their potential to infect others. CONCLUSION: Participants displayed confusion about transmission risks for hepatitis C, conflating blood awareness and hygiene health promotion messages. Participants' reliance on the symptom of jaundice to gauge serostatus places them at risk of transmitting and contracting HCV. Participants were confused about what a positive HCV diagnosis meant for their own health and their ability to infect others. Education is needed to debunk misconceptions about jaundice and clarify medical terms such as 'antibody' at the time of diagnosis. Further clarification of messages about injecting hygiene and blood awareness are also required. PMID- 15788092 TI - Optimising surgical management of elderly cancer patients. AB - BACKGROUND: Elderly population is on rise. It is an ethical dilemma how aggressive one should be when it comes to treat cancer in elderly. Presumed fear of increased postoperative morbidity and mortality has resulted in delivery of sub-optimal cancer surgery. METHODS: In this review article we visit physiology of the aged, tools available to assess surgical risks in oncogeriatric patients, and current practice in the management of common cancers encountered in surgical oncology, with the view of increasing awareness on optimising surgical management of senior patients with cancer. A pubmed search for cancer, surgery, elderly, was carried out. RESULTS: Cancer is on rise with increasing age predominantly affecting breast, gastrointestinal tract and lung. Increasingly more surgeons are offering surgery to elderly cancer patient but selection bias is prevalent. Available data reflect short and long-term outcome of cancer surgery in elderly is not greatly different to that of younger patient. Declining physiological reserve along with inability to respond adequately to physiological stress are salient age related changes. Comprehensive Geriatric Assessment (CGA) is not tested in surgical patient. There is need for a tool to define individualised operative risk. Preoperative assessment of cancer in elderly is designed to offer this information based on functional status of an individual utilising currently available tools of risk assessment. CONCLUSION: All elderly cancer patients should be offered optimal treatment depending on their functional status not on chronological age. Oncogeriatric patient would benefit from dedicated multidisciplinary approach. Recruitment of elderly cancer patients to more clinical trials is needed to enhance our knowledge and to offer optimum treatment to this unique subgroup. PMID- 15788094 TI - Gene expression analysis reveals that histone deacetylation sites may serve as partitions of chromatin gene expression domains. AB - BACKGROUND: It has been a long-term puzzle whether chromatin can be further divided into distinct gene expression domains. Because histone deacetylation affects chromatin structure, that in turn may affect the expression of nearby genes, histone deacetylation sites may act to partition chromatin into different gene expression domains. In this article, we explore the relationship between histone deacetylation sites and gene expression patterns on the genome scale using different data sources, including microarray data measuring gene expression levels, microarray data measuring histone deacetylation sites, and information on regulatory targets of transcription factors. RESULTS: Using 269 Saccharomyces cerevisiae microarray datasets, histone deacetylation datasets, and regulatory targets of transcription factors assembled from the Yeast Proteome Database and ChIP-chip data, we found that histone deacetylation sites can reduce the level of co-expression of neighboring genes. CONCLUSION: Histone deacetylation sites may serve as possible partition sites for chromatin domains and affect gene expression. PMID- 15788095 TI - Feature selection and nearest centroid classification for protein mass spectrometry. AB - BACKGROUND: The use of mass spectrometry as a proteomics tool is poised to revolutionize early disease diagnosis and biomarker identification. Unfortunately, before standard supervised classification algorithms can be employed, the "curse of dimensionality" needs to be solved. Due to the sheer amount of information contained within the mass spectra, most standard machine learning techniques cannot be directly applied. Instead, feature selection techniques are used to first reduce the dimensionality of the input space and thus enable the subsequent use of classification algorithms. This paper examines feature selection techniques for proteomic mass spectrometry. RESULTS: This study examines the performance of the nearest centroid classifier coupled with the following feature selection algorithms. Student-t test, Kolmogorov-Smirnov test, and the P-test are univariate statistics used for filter-based feature ranking. From the wrapper approaches we tested sequential forward selection and a modified version of sequential backward selection. Embedded approaches included shrunken nearest centroid and a novel version of boosting based feature selection we developed. In addition, we tested several dimensionality reduction approaches, namely principal component analysis and principal component analysis coupled with linear discriminant analysis. To fairly assess each algorithm, evaluation was done using stratified cross validation with an internal leave-one-out cross validation loop for automated feature selection. Comprehensive experiments, conducted on five popular cancer data sets, revealed that the less advocated sequential forward selection and boosted feature selection algorithms produce the most consistent results across all data sets. In contrast, the state-of-the-art performance reported on isolated data sets for several of the studied algorithms, does not hold across all data sets. CONCLUSION: This study tested a number of popular feature selection methods using the nearest centroid classifier and found that several reportedly state-of-the-art algorithms in fact perform rather poorly when tested via stratified cross-validation. The revealed inconsistencies provide clear evidence that algorithm evaluation should be performed on several data sets using a consistent (i.e., non-randomized, stratified) cross-validation procedure in order for the conclusions to be statistically sound. PMID- 15788097 TI - On geography and medical journalology: a study of the geographical distribution of articles published in a leading medical informatics journal between 1999 and 2004. AB - BACKGROUND: Studying the contribution of individual countries to leading journals in a given discipline can highlight which countries have the most impact on that discipline, and also give some idea about the geographical outreach of those journals. This paper examines the number of countries that contributed articles to one leading medical informatics journal, Medical Informatics & the Internet in Medicine, and the amount of their contributions between 1999 and the first half of 2004. METHODS: The PubMed citations of all indexed articles from the chosen journal (n = 128) were retrieved online (up to Volume 29, Number 2/June 2004 issue, the latest indexed issue as at 28 January 2005). The country of corresponding author's affiliation for each retrieved citation was recorded. The five-year-and-half corpus of abstracts retrieved from PubMed was further explored using MetaCarta Geographic Text Search http://www.metacarta.com/. RESULTS: The examined journal has an international outreach, with authors from 24 countries, spanning four continents, contributing to the journal during the studied period. The journal is dominated by a very large number of articles from Europe (81.25% of all articles counted in this study), and in particular from the UK (15.63%) and Greece (15.63%). There were no contributions from Africa or South America. CONCLUSION: A detailed discussion and interpretation of these results and ideas for future analyses are provided. MetaCarta can prove very useful as a bibliometric research tool. PMID- 15788096 TI - The "Statinth" wonder of the world: a panacea for all illnesses or a bubble about to burst. AB - After the introduction of statins in the market as effective lipid lowering agents, they were shown to have effects other than lipid lowering. These actions were collectively referred to as 'pleiotropic actions of statins.' Pleiotropism of statins formed the basis for evaluating statins for several indications other than lipid lowering. Evidence both in favour and against is available for several of these indications. The current review attempts to critically summarise the available data for each of these indications. PMID- 15788099 TI - Actinomycetes as host cells for production of recombinant proteins. AB - Actinomycetes (Actinobacteria) are highly attractive as cell factories or bioreactors for applications in industrial, agricultural, environmental, and pharmaceutical fields. Genome sequencing of several species of actinomycetes has paved the way for biochemical and structural analysis of important proteins and the production of such proteins as recombinants on a commercial scale. In this regard, there is a need for improved expression vectors that will be applicable to actinomycetes. Recent advancements in gene expression systems, knowledge regarding the intracellular environment, and identification and characterization of plasmids has made it possible to develop practicable recombinant expression systems in actinomycetes as described in this review. PMID- 15788098 TI - Heterologous expression of the filarial nematode alt gene products reveals their potential to inhibit immune function. AB - BACKGROUND: Parasites exploit sophisticated strategies to evade host immunity that require both adaptation of existing genes and evolution of new gene families. We have addressed this question by testing the immunological function of novel genes from helminth parasites, in which conventional transgenesis is not yet possible. We investigated two such novel genes from Brugia malayi termed abundant larval transcript (alt), expression of which reaches ~5% of total transcript at the time parasites enter the human host. RESULTS: To test the hypothesis that ALT proteins modulate host immunity, we adopted an alternative transfection strategy to express these products in the protozoan parasite Leishmania mexicana. We then followed the course of infection in vitro in macrophages and in vivo in mice. Expression of ALT proteins, but not a truncated mutant, conferred greater infectivity of macrophages in vitro, reaching 3-fold higher parasite densities. alt-transfected parasites also caused accelerated disease in vivo, and fewer mice were able to clear infection of organisms expressing ALT. alt-transfected parasites were more resistant to IFN-gamma induced killing by macrophages. Expression profiling of macrophages infected with transgenic L. mexicana revealed consistently higher levels of GATA-3 and SOCS-1 transcripts, both associated with the Th2-type response observed in in vivo filarial infection. CONCLUSION: Leishmania transfection is a tractable and informative approach to determining immunological functions of single genes from heterologous organisms. In the case of the filarial ALT proteins, our data suggest that they may participate in the Th2 bias observed in the response to parasite infection by modulating cytokine-induced signalling within immune system cells. PMID- 15788100 TI - Lumping or splitting: seeking the preferred areal unit for health geography studies. AB - BACKGROUND: Findings are compared on geographic variation of incident and late stage cancers across Connecticut using different areal units for analysis. RESULTS: Few differences in results were found for analyses across areal units. Global clustering of incident prostate and breast cancer cases was apparent regardless of the level of geography used. The test for local clustering found approximately the same locales, populations at risk and estimated effects. However, some discrepancies were uncovered. CONCLUSION: In the absence of conditions calling for surveillance of small area cancer clusters ('hot spots'), the rationale for accepting the burdens of preparing data at levels of geography finer than the census tract may not be compelling. PMID- 15788101 TI - Exogenous glycosaminoglycans coat damaged bladder surfaces in experimentally damaged mouse bladder. AB - BACKGROUND: Interstital cystitis is often treated with exogenous glycosaminoglycans such as heparin, chondroitin sulphate (Uracyst), hyaluronate (Cystistat) or the semi-synthetic pentosan polysulphate (Elmiron). The mechanism of action is presumed to be due to a coating of the bladder surface to replace the normally present chondroitin sulphate and heparan sulphate lost as a result of the disease. This study used fluorescent labelled chondroitin sulphate to track the distribution of glycosaminoglycans administered intravesically to mouse bladder that had been damaged on the surface. METHODS: The surfaces of mouse bladders were damaged by 3 mechanisms -- trypsin, 10 mM HCl, and protamine sulphate. Texas Red-labeled chondroitin sulphate was instilled into the bladders of animals with damaged bladders and controls instilled only with saline. Bladders were harvested, frozen, and sectioned for examination by fluorescence. RESULTS: The normal mouse bladder bound a very thin layer of the labelled chondroitin sulphate on the luminal surface. Trypsin- and HCl-damaged bladders bound the labelled chondroitin sulphate extensively on the surface with little penetration into the bladder muscle. Protamine produced less overt damage, and much less labelling was seen, presumably due to loss of the label as it complexed with the protamine intercalated into the bladder surface. CONCLUSION: Glycosaminoglycan administered intravesically does bind to damaged bladder. Given that the changes seen following bladder damage resemble those seen naturally in interstitial cystitis, the mechanisms proposed for the action of these agents is consistent with a coating of damaged bladder. PMID- 15788102 TI - Evolutionary cores of domain co-occurrence networks. AB - BACKGROUND: The modeling of complex systems, as disparate as the World Wide Web and the cellular metabolism, as networks has recently uncovered a set of generic organizing principles: Most of these systems are scale-free while at the same time modular, resulting in a hierarchical architecture. The structure of the protein domain network, where individual domains correspond to nodes and their co occurrences in a protein are interpreted as links, also falls into this category, suggesting that domains involved in the maintenance of increasingly developed, multicellular organisms accumulate links. Here, we take the next step by studying link based properties of the protein domain co-occurrence networks of the eukaryotes S. cerevisiae, C. elegans, D. melanogaster, M. musculus and H. sapiens. RESULTS: We construct the protein domain co-occurrence networks from the PFAM database and analyze them by applying a k-core decomposition method that isolates the globally central (highly connected domains in the central cores) from the locally central (highly connected domains in the peripheral cores) protein domains through an iterative peeling process. Furthermore, we compare the subnetworks thus obtained to the physical domain interaction network of S. cerevisiae. We find that the innermost cores of the domain co-occurrence networks gradually grow with increasing degree of evolutionary development in going from single cellular to multicellular eukaryotes. The comparison of the cores across all the organisms under consideration uncovers patterns of domain combinations that are predominately involved in protein functions such as cell-cell contacts and signal transduction. Analyzing a weighted interaction network of PFAM domains of yeast, we find that domains having only a few partners frequently interact with these, while the converse is true for domains with a multitude of partners. Combining domain co-occurrence and interaction information, we observe that the co-occurrence of domains in the innermost cores (globally central domains) strongly coincides with physical interaction. The comparison of the multicellular eukaryotic domain co-occurrence networks with the single celled of S. cerevisiae (the overlap network) uncovers small, connected network patterns. CONCLUSION: We hypothesize that these patterns, consisting of the domains and links preserved through evolution, may constitute nucleation kernels for the evolutionary increase in proteome complexity. Combining co-occurrence and physical interaction data we argue that the driving force behind domain fusions is a collective effect caused by the number of interactions and not the individual interaction frequency. PMID- 15788103 TI - Frequent detection of worm movements in onchocercal nodules by ultrasonography. AB - BACKGROUND: Ultrasonography (USG) is known to be a suitable tool for diagnosis in lymphatic filariasis as the adult filarial nematode Wuchereria bancrofti in scrotal lymphatic vessels of infected men can be detected by the characteristic pattern of movement, the Filaria Dance Sign. In onchocerciasis, moving adult worms have not yet been demonstrated by USG. In addition the verification of drug effects on living adult Onchocerca volvulus filariae in trials is hampered by the lack of tools for longitudinal observation of alterations induced by potentially macrofilaricidal drugs in vivo. The present study was carried out to determine the frequency of detection of moving adult filariae of O. volvulus by USG. METHODS: In an endemic region for onchocerciasis in Ghana, 61 patients infected with onchocerciasis were recruited by palpation and onchocercomas examined by USG using an ultrasound system equipped with a 7.5 - 10 MHz linear transducer. Onchocercomas were recorded on videotape and evaluated with regard to location, number and size, as well as to movements of adult filariae. RESULTS: In the 61 patients 303 onchocercomas were found by palpation and 401 onchocercomas were detected by USG. In 18 out of 61 patients (29.5%), altogether 22 nodules with moving adult O. volvulus filariae were detected and are presented in animated ultrasound images as mp-4 videos. CONCLUSION: Ultrasonographical examinations of onchocercomas where living adult filariae can be displayed may serve as a new tool for the longitudinal observation in vivo of patients with onchocerciasis undergoing treatment and as an adjunct to histological evaluation. PMID- 15788104 TI - Identification to the species level of Lactobacillus isolated in probiotic prospecting studies of human, animal or food origin by 16S-23S rRNA restriction profiling. AB - BACKGROUND: The accurate identification of Lactobacillus and other co-isolated bacteria during microbial ecological studies of ecosystems such as the human or animal intestinal tracts and food products is a hard task by phenotypic methods requiring additional tests such as protein and/or lipids profiling. RESULTS: Bacteria isolated in different probiotic prospecting studies, using de Man, Rogosa and Sharpe medium (MRS), were typed at species level by PCR amplification of 16S-23S rRNA intergenic spacers using universal primers that anneal within 16S and 23S genes, followed by restriction digestion analyses of PCR products. The set of enzymes chosen differentiates most species of Lactobacillus genus and also co-isolated bacteria such as Enterococcus, Streptococcus, Weissella, Staphylococcus, and Escherichia species. The in silico predictions of restriction patterns generated by the Lactobacillus shorter spacers digested with 11 restriction enzymes with 6 bp specificities allowed us to distinguish almost all isolates at the species level but not at the subspecies one. Simultaneous theoretical digestions of the three spacers (long, medium and short) with the same set of enzymes provided more complex patterns and allowed us to distinguish the species without purifying and cloning of PCR products. CONCLUSION: Lactobacillus isolates and several other strains of bacteria co-isolated on MRS medium from gastrointestinal ecosystem and fermented food products could be identified using DNA fingerprints generated by restriction endonucleases. The methodology based on amplified ribosomal DNA restriction analysis (ARDRA) is easier, faster and more accurate than the current methodologies based on fermentation profiles, used in most laboratories for the purpose of identification of these bacteria in different prospecting studies. PMID- 15788105 TI - Interpretation of the post-surgical Somatostatin Receptor Scintigram of a Primary Neuroendocrine Tumor of the Thymus: a case report and literature review. AB - A case of a thymic neuroendocrine tumor and the interpretation problems in a post surgical Somatostatin Receptor Scintigraphy are presented. In a 53-year-old man with superior vena cava obstruction syndrome an atypical carcinoid of the thymus (neuroendocrine carcinoma of intermediate grade 2), was found at surgery.During his first year of follow-up a Somatostatin Receptor Scintigraphy was recommended. An area of abnormal concentration of the radiopharmaceutical was revealed in the mediastinum at this time.A thorough understanding of the mechanisms of the radiopharmaceutical uptake and of the various clinical settings in which uptake can occur are essential for a proper evaluation of the scintigraphic findings and result in the optimal use of this valuable modality.The literature review provides an overview of this rare type of tumor and insight into the clinical significance of Somatostatin Receptor Scintigraphy. PMID- 15788106 TI - Wildfire: distributed, Grid-enabled workflow construction and execution. AB - BACKGROUND: We observe two trends in bioinformatics: (i) analyses are increasing in complexity, often requiring several applications to be run as a workflow; and (ii) multiple CPU clusters and Grids are available to more scientists. The traditional solution to the problem of running workflows across multiple CPUs required programming, often in a scripting language such as perl. Programming places such solutions beyond the reach of many bioinformatics consumers. RESULTS: We present Wildfire, a graphical user interface for constructing and running workflows. Wildfire borrows user interface features from Jemboss and adds a drag and-drop interface allowing the user to compose EMBOSS (and other) programs into workflows. For execution, Wildfire uses GEL, the underlying workflow execution engine, which can exploit available parallelism on multiple CPU machines including Beowulf-class clusters and Grids. CONCLUSION: Wildfire simplifies the tasks of constructing and executing bioinformatics workflows. PMID- 15788107 TI - The influence of moderate red wine consumption on antioxidant status and indices of oxidative stress associated with CHD in healthy volunteers. AB - The effects of moderate red wine consumption on the antioxidant status and indices of lipid peroxidation and oxidative stress associated with CHD were investigated. A randomised, controlled study was performed with twenty free living healthy volunteers. Subjects in the red wine group consumed 375 ml red wine daily for 2 weeks. We measured the total concentration of phenolics and analysed the individual phenolics in the wine and plasma by HPLC with tandem MS. The antioxidant capacity of plasma was measured with electron spin resonance spectroscopy while homocysteine and fasting plasma lipids were also determined. The production of conjugated dienes and thiobarbituric acid-reactive substances (TBARS) were measured in Cu-oxidised LDL. Plasma total phenolic concentrations increased significantly after 2 weeks of daily red wine consumption (P< or =0.001) and trace levels of metabolites, mainly glucuronides and methyl glucuronides of (+)-catechin and (-)-epicatechin, were detected in the plasma of the red wine group. These flavan-3-ol metabolites were not detected in plasma from the control group. The maximum concentrations of conjugated dienes and TBARS in Cu-oxidised LDL were reduced (P< or =0.05) and HDL cholesterol concentrations increased (P< or =0.05) following red wine consumption. The findings from the present study provide some evidence for potential protective effects of moderate consumption of red wine in healthy volunteers. PMID- 15788108 TI - Vitamins A and E: metabolism, roles and transfer to offspring. AB - Vitamins A and E are essential, naturally occurring, fat-soluble nutrients that are involved in several important biological processes such as immunity, protection against tissue damage, reproduction, growth and development. They are extremely important during the early stages of life and must be transferred adequately to the young during gestation and lactation. The present article presents an overview of their biological functions, metabolism and dynamics of transfer to offspring in mammals. Among other topics, the review focuses on the biochemical aspects of their intestinal absorption, blood transport, tissue uptake, storage and catabolism. It also describes their different roles as well as their use as preventive and therapeutic agents. Finally, the mechanisms involved in their transfer during gestation and lactation are discussed. PMID- 15788109 TI - Dissociation of the glycaemic and insulinaemic responses to whole and skimmed milk. AB - In most carbohydrate-containing foods, the blood insulin response is predictable and is closely linked to the food's glycaemic index (GI). A single study, examining whole milk and fermented milk products made from whole milk, recently reported a large dissociation between the GI and insulinaemic index (II) in healthy normal adults. Because the fat component of a food may influence the GI and II, it is unclear if a similar dissociation may exist for skimmed milk in normal adults. We determined the GI and II of both skimmed and whole milk in nine healthy, male (n 6) and female (n 3) subjects (23.6 (sd 1.4) years). No significant (P>0.05) differences existed between GI and II for skimmed and whole milks. Significant (P<0.05) differences were observed between the actual and predicted areas under the insulin curves for both skimmed milk (predicted 1405 (sd 289) pmol x min/l; actual 6152 (sd 1177) pmol x min/l) and whole milk (predicted 1564 (sd 339) pmol x min/l; actual 5939 (sd 1095) pmol x min/l). Consequently, a large and similar dissociation of the GI and II existed for both whole milk (42 (sd 5) and 148 (sd 14)) and skimmed milk (37 (sd 9) and 140 (sd 13)). It is concluded that the dissociation of the GI and II in milk is not related to its fat content. PMID- 15788110 TI - The beneficial effect of a diet with low glycaemic index on 24 h glucose profiles in healthy young people as assessed by continuous glucose monitoring. AB - Elevated postprandial glycaemia has been linked to CVD in a number of different epidemiological studies involving predominantly non-diabetic volunteers. The MiniMed continuous glucose monitor, which measures blood glucose every 5 min, over a 24 h period, was used to investigate changes in blood glucose readings before and after instigating a diet with low glycaemic index (GI) for 1 week in free-living healthy individuals. Nine healthy people (age 27 (SEM 1.3) years, BMI 23.7 (SEM 0.7) kg/m2, one male, eight females) completed the study. A reduction in GI (59.7 (SEM 2) v. 52.1 (SEM 2), P<0.01) occurred in all nine subjects while energy and other macronutrients remained constant. A significant reduction was also observed in fasting glucose at 06.00 hours (5.4 (SEM 0.2) v. 4.4 (SEM 0.3) mmol/l, P<0.001), mean glucose (5.6 (SEM 0.2) v. 5.1 (SEM 0.2) mmol/l, P=0.004), area under the 24 h glucose curve (8102 (SEM 243) v. 750 (SEM 235) mmol/l per min, P=0.004) and area under the overnight, 8 h glucose curve (2677 (SEM 92) v. 2223 (SEM 121) mmol/l per min, P=0.01). The present study provides important data on how a simple adjustment to the diet can improve glucose profiles that, if sustained in the long term, would be predicted from epidemiological studies to have a favourable influence on CVD. PMID- 15788111 TI - Effect of arginine deficiency on arginine-dependent post-translational protein modifications in mice. AB - Transgenic mice that overexpress arginase-I in their small-intestinal enterocytes suffer from a pronounced, but selective decrease in circulating arginine levels during the suckling period, resulting in impaired growth and development of hair, muscle and immune system. In the present study, we tested the hypothesis that the arginine-deficiency phenotype is caused by arginine-specific post-translational modifications, namely, an increase in the degree of mono-ADP-ribosylation of proteins because of reduced competition by free arginine residues and/or an increase in protein-tyrosine nitration because of an increased O2- production by NO synthases in the presence of limiting amounts of arginine. Arginine ADP ribosylation and tyrosine nitration of proteins in the affected organs were assayed by Western blot analysis, using specific anti-ADP-ribosylarginine and protein-nitrotyrosine antisera. The composition of the group of proteins that were preferentially arginine ADP-ribosylated or tyrosine-nitrated in the respective organs was strikingly similar. Arginine-deficient mice differed from their controls in a reduced ADP-ribosylation of a 130 kDa and a 65 kDa protein in skin and an increased protein nitration of an 83 kDa protein in bone marrow and a 250 kDa protein in spleen. Since only 20 % of the visualised proteins were differentially modified in a subset of the affected organs, our findings appear to rule out these prominent arginine-dependent post-translational protein modifications as mediators of the characteristic phenotype of severely arginine deficient mice. PMID- 15788112 TI - Effects of subthalamic nucleus deep brain stimulation and levodopa on energy production rate and substrate oxidation in Parkinson's disease. AB - Patients with Parkinson's disease (PD) often lose weight, but after subthalamic nucleus deep brain stimulation (STN-DBS), they gain weight. We compared daily energy intake (DEI), resting energy expenditure (REE) and substrate oxidation rates (measured by indirect calorimetry) in nineteen STN-DBS-treated patients (Group S), thirteen others on pharmacologic treatment by levodopa (Group L) and eight control subjects. We also determined the acute effects of STN-DBS and levodopa on REE and substrate oxidation rates. STN-DBS treated patients gained 9.7 (SEM 7.1) kg after surgery, whereas patients on pharmacologic treatment lost 3.8 (SEM 10.0) kg since diagnosis. In STN-DBS-treated patients, REE (-16.5 %; P<0.001), lipid oxidation (-27 %; P<0.05) and protein oxidation (-46 %; P<0.05) were decreased, whereas glucose oxidation was elevated (+81 %; P<0.05) as compared to patients on pharmacologic treatment. Levodopa acutely reduced REE ( 8.3 %; P<0.05) and glucose oxidation (-37 %; P<0.01) with a slight hyperglycaemic effect (after levodopa challenge: 5.6 (SEM 0.8) v. before levodopa challenge: 5.3 (SEM 0.6) mmol/l; P<0.01). Switching 'on' STN-DBS acutely reduced REE (-17.5 %; P<0.01) and lipid oxidation (-24 %; P<0.001) 30 min after starting stimulation. Fasting glycaemia was slightly but significantly reduced (5.4 (SEM 1.4) v. 5.5 (SEM 1.3) mmol/l; P<0.01). After STN-DBS, the normalization of REE and the reduction in lipid and protein oxidation contribute to the restoration of weight. As levodopa decreases glucose oxidation, the reduction in daily dose of levodopa in STN-DBS-treated patients helps prevent the effect of weight gain on glycaemia. PMID- 15788113 TI - Water loss as a function of energy intake, physical activity and season. AB - Although water is an important nutrient, there are no recommended intake values. Here, water intake, energy intake, physical activity and water loss was measured over 1 week in summer and in winter. Subjects were healthy volunteers, forty-two women and ten men, mean age of 29 (SD 7) years and mean BMI 21.8 (SD 2.2) kg/m2. Water intake was measured with a 7 d food and water record. Physical activity level (PAL) was observed as the ratio of total energy expenditure, as measured with doubly labelled water, to resting energy expenditure as measured in a respiration chamber. Water loss was measured with the deuterium elimination method. Water loss was highly reproducible and ranged from 0.20 to 0.35 l/MJ, independent of season and activity level, with higher values in women. Water loss was related to water and energy intake in summer (r 0.96, P<0.0001 and r 0.68, P<0.001, respectively) as well as in winter (r 0.98, P<0.0001 and r 0.63, P<0.01, respectively). Water loss was, for men, higher in subjects with a higher physical activity in summer (r 0.94, P<0.0001) and in winter (r 0.70, P<0.05). Normalizing water loss for differences in energy expenditure by expressing water loss in litres per MJ resulted in the same value for men in summer and winter. For women, physical activity-adjusted values of water loss were higher, especially in summer. In men, water turnover was determined by energy intake and physical activity, while seasonal effects appeared through energy expenditure. Women showed a higher water turnover that was unrelated to physical activity. PMID- 15788114 TI - Partitioning of limiting protein and energy in the growing pig: description of the problem, possible rules and their qualitative evaluation. AB - A core part of any animal growth model is how it predicts the partitioning of dietary protein and energy to protein and lipid retention for different genotypes at different degrees of maturity. Rules of partitioning need to be combined with protein and energy systems to make predictions. The animal needs describing in relation to its genotype, live weight and, possibly, body composition. Some existing partitioning rules will apply over rather narrow ranges of food composition, animal and environment. Ideally, a rule would apply over the whole of the possible experimental space (scope). The live weight range over which it will apply should at least extend beyond the 'slaughter weight range', and ideally would include the period from the start of feeding through to maturity. Solutions proposed in the literature to the partitioning problem are described in detail and criticised in relation to their scope, generality and economy of parameters. They all raise the issue, at least implicitly, of the factors that affect the net marginal efficiency of using absorbed dietary protein for protein retention. This is identified as the crucial problem to solve. A problem identified as important is whether the effects of animal and food composition variables are independent of each other or not. Of the rules in the literature, several could be rejected on qualitative grounds. Those rules that survived were taken forward for further critical and quantitative analysis in the companion paper. PMID- 15788115 TI - Partitioning of limiting protein and energy in the growing pig: testing quantitative rules against experimental data. AB - Literature solutions to the problem of protein and energy partitioning in the growing pig are quantitatively examined. Possible effects of live weight, genotype and food composition on the marginal response in protein retention to protein and energy intakes, on protein and energy-limiting foods are quantified. No evidence was found that the marginal response in protein retention to ideal protein supply, when protein intake is limiting, is affected by live weight, genotype or environmental temperature. There was good evidence that live weight does not affect the marginal response in protein retention to energy intake when protein intake is not limiting. Limited data for different genotypes suggested no effects on this response. A general quantitative partitioning rule is proposed that has two key parameters; e(p)* (the maximum marginal efficiency for retaining the first limiting amino acid) and R* (the maximum value of R, the energy to protein ratio of the food, MJ metabolisable energy (ME)/kg digestible crude protein (DCP), when e(p)* is just achieved). When R or =20 years in the National Health and Nutrition Examination Survey 1999-2000. Alpha-tocopherol concentration was inversely associated with glucose concentration (beta per mmol/l=-0.01064, SE 0.00356, P=0.004) after adjusting for age, sex, race or ethnicity, education, smoking status, concentrations of total cholesterol and triacylglycerols, systolic blood pressure, waist circumference, alcohol use, physical activity, time watching television or videos or using a computer, and use of vitamin/mineral/dietary supplements. Among 659 participants who did not report using supplements, this association was no longer significant whereas the concentration of alpha-tocopherol was inversely associated with concentration of C-peptide (beta per mmol/l=-0.01121, SE 0.00497, P=0.024). Gamma-tocopherol concentration was positively associated with concentration of glucose (beta per mmol/l=0.09169, SE 0.02711, P=0.001) and glycosylated haemoglobin (beta per mmol/l=0.04954, SE 0.01284, P<0.001), but not insulin or C-peptide. The relationships between physiologic concentrations of the various forms of vitamin E and measures of glucose intolerance deserve additional investigation. PMID- 15788119 TI - Antioxidant capacity of vegetables, spices and dressings relevant to nutrition. AB - Vegetables are the most important sources of phenolics in the Mediterranean diet. Phenolics, especially flavonoids, are suggested as being essential bioactive compounds providing health benefits. In this study, twenty-seven vegetables, fifteen aromatic herbs and some spices consumed in Central Italy (the Marches region) were studied to reveal total phenolic, flavonoid and flavanol content as well as their antioxidant capacity measured by the oxygen radical absorbance capacity (ORAC) method. A comparison in terms of antioxidant capacity was made between different salads, as well as between salads to which aromatic herbs had been added. Lemon balm and marjoram at a concentration of 1.5 % w/w increased by 150 % and 200 % respectively the antioxidant capacity of a salad portion. A 200 g portion of a salad enriched with marjoram corresponded to an intake of 200 (SD 10) mg phenolics and 4000 (SD 300) ORAC units (micromol Trolox equivalents). Olive oils and wine or apple vinegars were the salad dressings that provided the highest increase in antioxidant capacity. Among the spices tested, cumin and fresh ginger made the most significant contribution to the antioxidant capacity. The results are useful in surveying the antioxidant parameters of vegetables, herbs and spices produced and consumed in our geographical area as well as in quantifying the daily intake of phenolics and ORAC units. The results can be used in public health campaigns to stimulate the consumption of vegetables able to provide significant health protection in order to prevent chronic diseases. PMID- 15788120 TI - Effects of arginine-containing total parenteral nutrition on N balance and phagocytic activity in rats undergoing a partial gastrectomy. AB - The present study investigated the effect of arginine (Arg)-containing parenteral nutrition on phagocytic activity to elucidate the possible roles of Arg in the secretion of anabolic hormones and N balance in rats undergoing gastrectomy. Rats were divided into two experimental groups and received total parenteral nutrition (TPN). The TPN solutions were isonitrogenous and identical in nutrient compositions except for differences in amino acid content. One group received conventional TPN, the other group replaced 2 % of the total energy as Arg. After receiving TPN for 3 d, one-third of the rats in each experimental group were killed as the baseline group. The remaining rats underwent a partial gastrectomy and were killed 1 or 3 d after surgery. The results showed that there were no differences in N balance, plasma growth hormone and insulin-like growth factor-1 levels between the two groups before or after surgery. The phagocytic activity of peritoneal macrophages was higher in the Arg group than in the control group 1 d after surgery. There were no differences in the phagocytic activities of blood polymorphonuclear neutrophils between the two groups at various time points. TNF alpha levels in peritoneal lavage fluid were lower in the Arg group than in the control group on post-operative day 3. These results suggest that parenterally infused Arg enhances phagocytic activity and reduces the production of inflammatory mediators at the site of injury. However, Arg supplementation did not influence the secretion of anabolic hormones nor N balance in rats with a partial gastrectomy. PMID- 15788121 TI - Appetite sensations as a marker of overall intake. AB - The aim of this study was to evaluate the clinical utility of appetite sensations to characterize individual overall energy intake. A group of men (n 28) and women (n 23) was recruited to record their 'desire to eat', 'hunger', 'fullness' and 'prospective food consumption' (PFC) on visual analogue scales before a standardized meal test, immediately after and every 10 min for a period of 1 h after the meal. The 1 h post-meal area under the curve (1 h AUC) and the satiety quotient (SQ) were calculated for all appetite sensations. In a second visit, all participants were invited to eat three meals in order to measure total energy intake (TEI) and food preferences. Metabolic rate (MR) was also assessed to derive daily relative energy intake (REI) by subtracting this variable from TEI (TEI-MR=REI). The Three-Factor Eating Questionnaire scores were also calculated for all participants. One h AUC for fullness was the appetite sensation most strongly associated with TEI and REI (r-0.42, P< or =0.003 and r-0.32, P< or =0.05, respectively). SQ for fullness was the only predictor of TEI and REI (r 0.42, P< or =0.0003 and r-0.30, P< or =0.05, respectively). Restraint, disinhibition and hunger scores were not associated with appetite sensation variables. These results suggest that the fullness dimension seems to be a useful appetite sensation to predict long-term TEI and REI. Thus, assessment of appetite sensation such as fullness in response to a fixed load may be useful to evaluate individual overall energy intake. PMID- 15788122 TI - Additional protein intake limits weight regain after weight loss in humans. AB - Since long-term weight maintenance (WM) is a major problem, interventions to improve WM are needed. The aim of the study was to investigate whether the addition of protein to the diet might limit weight regain after a weight loss of 5-10 % in overweight subjects. In a randomised parallel study design, 113 overweight subjects (BMI 29.3 (SD 2.5) kg/m2); age 45.1 (SD 10.4) years) followed a very-low-energy diet for 4 weeks, after which there was a 6-month period of WM. During WM, subjects were randomised into either a protein group or a control group. The protein group received 30 g/d protein in addition to their own usual diet. During the very-low-energy diet, no differences were observed between the groups. During WM, the protein group showed a higher protein intake (18 % v. 15 %; P<0.05), a lower weight regain (0.8 v. 3.0 kg; P<0.05), a decreased waist circumference (-1.2 (SD 0.7) v. 0.5 (SD 0.5 ) cm; P<0.05) and a smaller increase in respiratory quotient (0.03 (SD 0.01) v. 0.07 0.01; (SD/)P <0.05) compared with the control group. Weight regain in the protein group consisted of only fat-free mass, whereas the control group gained fat mass as well. Satiety in the fasted state before breakfast increased significantly more in the protein group than in the control group. After 6 months follow-up, body weight showed a significant group x time interaction. A protein intake of 18 % compared with 15 % resulted in improved WM in overweight subjects after a weight loss of 7.5 %. This improved WM implied several factors, i.e. improved body composition, fat distribution, substrate oxidation and satiety. PMID- 15788123 TI - Interpreting serological studies on severe acute respiratory syndrome: defining the reference. PMID- 15788124 TI - Evaluation by indirect immunofluorescent assay and enzyme linked immunosorbent assay of the dynamic changes of serum antibody responses against severe acute respiratory syndrome coronavirus. AB - BACKGROUND: Severe acute respiratory syndrome coronavirus (SARS-CoV) is a newly emerging virus that gives rise to SARS patients with high rates of infectivity and fatality. To study the humoral immune responses to SARS-CoV, the authors evaluated IgG and IgM specific antibodies in patients' sera. METHODS: Two methods, enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent assay (IFA), were used to detect specific serum IgG and IgM against SARS-CoV in 98 SARS patients and 250 controls consisting of patients with pneumonia, health care professionals and healthy subjects. The serum antibody profiles were investigated at different times over one and a half years in 18 of the SARS patients. RESULTS: The sensitivity and specificity of ELISA for detecting IgG against SARS-CoV were 100.0% and 97.2% and for IgM 89.8% and 97.6% respectively; the figures using IFA for IgG were 100.0% and 100.0% and for IgM 81.8% and 100.0% respectively. During the first seven days of the antibodies trace test, no IgG and IgM were detected, but on day 15, IgG response increased dramatically, reaching a peak on day 60, remaining high up to day 180 and decreasing gradually until day 540. On day 15, IgM was detected, rapidly reached a peak, then declined gradually until day 180 when IgM was undetectable. CONCLUSION: The detection of antibodies against SARS virus is helpful in the clinical diagnosis of SARS. PMID- 15788125 TI - Severe acute respiratory syndrome coronavirus persistence in Vero cells. AB - BACKGROUND: Several coronaviruses establish persistent infections in vitro and in vivo, however it is unknown whether persistence is a feature of the severe acute respiratory syndrome coronavirus (SARS-CoV) life cycle. This study was conducted to investigate viral persistence. METHODS: We inoculated confluent monolayers of Vero cells with SARS-CoV at a multiplicity of infection of 0.1 TCID50 and passaged the remaining cells every 4 to 8 days for a total of 11 passages. Virus was titrated at each passage by limited dilution assay and nucleocapsid antigen was detected by Western blot and immunofluoresence assays. The presence of viral particles in passage 11 cells was assessed by electron microscopy. Changes in viral genomic sequences during persistent infection were examined by DNA sequencing. RESULTS: Cytopathic effect was extensive after initial inoculation but diminished with serial passages. Infectious virus was detected after each passage and viral growth curves were identical for parental virus stock and virus obtained from passage 11 cells. Nucleocapsid antigen was detected in the majority of cells after initial inoculation but in only 10%-40% of cells at passages 2-11. Electron microscopy confirmed the presence of viral particles in passage 11 cells. Sequence analysis at passage 11 revealed fixed mutations in the spike (S) gene and ORFs 7a-8b but not in the nucleocapsid (N) gene. CONCLUSIONS: SARS-CoV can establish a persistent infection in vitro. The mechanism for viral persistence is consistent with the formation of a carrier culture whereby a limited number of cells are infected with each round of virus replication and release. Persistence is associated with selected mutations in the SARS-CoV genome. This model may provide insight into SARS-related lung pathology and mechanisms by which humans and animals can serve as reservoirs for infection. PMID- 15788126 TI - Dominant cagA/vacA genotypes and coinfection frequency of H. pylori in peptic ulcer or chronic gastritis patients in Zhejiang Province and correlations among different genotypes, coinfection and severity of the diseases. AB - BACKGROUND: Almost half of the world's population suffer from the Helicobacter pylori (H. pylori) infection, but only some individuals develop gastric diseases with clinical symptoms. One reason for the phenomenon may be the different pathogenicity of infected H. pylori strains. The presence of cytotoxin-associated gene A (cagA) and expression of vacuolating cytotoxin activity encoded by vacuolating cytotoxin gene A (vacA) are considered the two major virulent markers of H. pylori. The aim of this study was to detect dominant cagA/vacA genotypes and coinfection frequency of H. pylori in patients with peptic ulceration (PU) or chronic gastritis (CG), and to determine correlations among different cagA/vacA genotypes, coinfection and severity of the diseases. METHODS: For each of 139 patients in Zhejiang Province who had been diagnosed as PU or CG based on clinical symptoms and gastroscopy, two gastric biopsy specimens (one from antrum and the other from corpus) for H. pylori isolation were taken by two different disinfected biopsy forceps. One hundred and fifty-six H. pylori strains were isolated from both the antrum and corpus biopsy specimens of 78 patients (36 PU and 42 CG). PCRs were performed to detect cagA genes, and signal (s) and middle (m) regions of vacA genes in the H. pylori isolates. The amplified fragments of dominant vacA gene s and m subtypes from representative H. pylori isolates were sequenced after TA cloning. Dominant cagA/vacA genotypes of the H. pylori isolates, coinfection frequency and correlations among the different genotypes, coinfection and severity of the diseases were determined. RESULTS: Of the H. pylori strains isolated from the antrum specimens, 96.2% were cagA gene positive, as were 97.4% of the H. pylori strains isolated from the corpus specimens. Only one s region subtype (s1a) and four m region subtypes m1, m2, m1b and m1b-m2 of vacA gene were found. The proportions of vacA gene subtypes s1a/m1, s1a/m2, s1a/m1b and s1a/m1b-m2 in the 83 strains isolated from the antrum specimens were 7.2%, 61.5%, 30.1% and 1.2%, respectively, while those in the other 84 strains isolated from the corpus specimens were 9.5%, 58.3%, 28.6% and 3.6%, respectively. s1a/m2 (58.3% vs 30.1%, chi(2) = 13.47, P < 0.01) and then s1a/m1b (28.6% vs 9.5%, chi(2) = 9.88, P < 0.01) were the dominant vacA gene subtypes in the H. pylori isolates. The dominant H. pylori genotype was cagA + s1a/m2 (59.0% from antrum specimens and 57.1% from corpus specimens), and followed by cagA + s1a/m1b (28.9% from antrum specimens and 27.4% from corpus specimens). Sixteen of 78 patients (20.5%) were infected with two or three H. pylori strains with different genotypes. However, no statistically significant differences among cagA occurrence, the different vacA subtypes and PU or CG could be found (each P > 0.05). Similarities of the nucleotide sequences from vacA gene s region PCR products of six isolates and from vacA gene m region PCR products of four isolates were 93.2% to 98.3% and 93.8% to 97.6%, respectively, compared to the reported corresponding sequences. CONCLUSIONS: The dominant genotypes of H. pylori in PU or CG patients in Zhejiang area may be cagA + s1a/m2 and cagA + s1a/m1b. Numerous coinfections with different H. pylori strains in PU or CG patients indicate diversity of the infected H. pylori origins. s and m regions of vacA gene from different H. pylori isolates show high nucleotide sequence similarities. cagA gene positive rate, different vacA gene subtypes and coinfection with different H. pylori strains are not closely associated with severity of the diseases. PMID- 15788127 TI - Study on the pathophysiologic basis of classification of 'spleen' deficiency in chronic gastritis. AB - BACKGROUND: Most of the studies on traditional Chinese medicine (TCM) 'spleen' deficiency syndrome in the recent 30 years were conducted only on the basis of single functional index, neglecting the study on the pathophysiologic internal relationship between spleen deficiency syndrome and gastric diseases in modern medicine. But it was at the subcellular molecular biological level that we explored the pathophysiologic basis of classification of spleen deficiency in chronic gastritis by detecting the bioactive substances in gastric mucosa nuclei and mitochondria. METHODS: By means of optical microscope, scanning electron microscope (SEM), transmission electron microscopy (TEM) and histochemical staining, we conducted histopathological, subcellular ultrastructural analysis and nuclei and mitochondrial ultrastructural analysis of gastric mucosa of 188 spleen deficiency patients and of 42 voluntary blood donors. At the same time, bioactive substances were measured by means of X-ray energy dispersive analysis system (EDAX) image analysis system, radioimmunoassay method and chemiluminescence method. RESULTS: The content of cAMP, superoxide dismutase (SOD), Zn and Cu in gastric mucosa, and the content of Zn and Cu in mitochondria decreased progressively in order of groups: healthy control (HC), spleen Qi deficiency without organic lesion (F-SQD), spleen Yang deficiency without organic lesion (F-SyangD), disease without symptoms group, spleen Qi deficiency with organic lesion (G-SQD), spleen Yang deficiency with organic lesion (G-SyangD), spleen Yin deficiency (SyinD) and spleen deficiency with Qi stagnation (SDQS), chronic spleen deficiency gastritis (CSG) and chronic atrophic gastritis (CAG); decreased in order of HC, intestinal metaplasia (IM)Ia, IMIb, IMIIa and IMIIb, P < 0.05. The content of DNA, Zn and Cu in nuclei progressively increased in order mentioned above, P < 0.05. CONCLUSIONS: The quantitative changes of gastric mucosal cAMP, SOD, Zn, Cu, of mitochondrial Zn, Cu and of nuclear DNA, Zn and Cu are not only the substance base on which the lesion of gastric mucosa tissue structure occurs, but also the substance base on which spleen deficiency is classified. G-SQD and G-SyangD were more likely to be found in low-grade or middle-grade CSG and CAG, while SyinD and SDQS in middle-grade or high-grade CSG, CAG and IMIIb. PMID- 15788128 TI - Graft-versus-leukemia effects from donor lymphocyte infusion after nonmyeloablative allogeneic bone marrow transplantation in mice. AB - BACKGROUND: Nonmyeloablative allogeneic bone marrow transplantation has been used since the 1990s as a new hematological stem cell transplantation strategy for treating hematological diseases. The purpose of this study was to explore the graft-versus-leukemia (GVL) effects of donor lymphocyte infusions (DLIs) after nonmyeloablative allogeneic bone marrow transplantations, while assessing the declines in treatment-associated morbidity, mortality, and graft-versus-host disease (GVHD). METHODS: A total of 615 (H-2k) mice were injected with L615 tumor cells and received 500 cGy (60Co gamma-ray) irradiation three days later, followed by an allogeneic bone marrow transplantation (allo-BMT). The allo-grafts consisted of 3 x 10(7) bone marrow cells and 1 x 10(7) spleen cells from BALB/C (H-2d) donor mice. Two days after the allo-BMT, the recipient mice were given 200 mg/kg of cyclophosphamide. Subsequently, recipient mice were infused with either donor spleen cells (2 x 10(7)) on day 14 or 21, or donor spleen cells (5 x 10(7)) pretreated with hydrocortisone and cyclosporin A (CsA) in vitro on day 14 post BMT. RESULTS: The median survival time of mice that received DLI on day 21 and pretreated DLI on day 14 post-BMT was longer than that of controls and the day 14 DLI group (P < 0.01). No evidence of severe GVHD was observed in the day 21 DLI group nor in the day 14 treated DLI group. Mixed chimerism was confirmed in the day 14 DLI group, the day 14 treated DLI group, and the day 21 DLI group on the thirteenth day post-transplantation; full donor chimerism was observed two weeks after DLI. CONCLUSION: Donor lymphocyte infusion after nonmyeloablative bone marrow transplantation may reduce transplantation-associated morbidity and mortality while strengthening graft-versus-leukemia effects. PMID- 15788130 TI - Induction of apoptosis by homoharringtonine in G1 phase human chronic myeloid leukemic cells. AB - BACKGROUND: Homoharringtonine (HHT) is a cephalotaxine ester derived from an evergreen tree found wildely throughout southern China, which has antileukemic activities against a variety of acute myeloid leukemic cells. For the sake of illustrating the mechanisms of HHT in the treatment of leukemia, we assessed the effect of HHT on the apoptosis of human chronic myeloid leukemic cell line K562. METHODS: The apoptosis of K562 cells induced by HHT was analyzed by transmission electron microscopy, agarose gel electrophoresis of DNA, flow cytometry and terminal deoxyribonucleotidyl transferase-mediated dUTP-biotin nick labeling. RESULTS: Characteristic apoptosis-related features emerged in K562 cells after exposed to HHT at a concentration 0.05-100 microg/ml. Transmission electron microscopy of HHT treated K562 cells displayed chromatin condensation and aggregation under the nuclear membrane, nuclear fragmentation and apoptosis body formation. Typical DNA ladder in agarose gel electrophoresis was observed in the cells exposed to HHT. The cell cycle analysis measured by flow cytometry showed G1 phase cells decreased with the increase of S phase cells while apoptosis was induced by HHT in K562 cells. The percentage of apoptotic cells in K562 cells treated with 50 microg/ml of HHT decreased significantly when pretreated with 1 microg/ml of cycloheximide, 0.05 microg/ml of Actinomycin D respectively. CONCLUSIONS: HHT has apoptotic effects on K562 cells. The HHT induced apoptosis mainly of the cells in G1 phase and this process required RNA transcription and protein synthesis. PMID- 15788129 TI - Inhibiting the expression of CD28 costimulatory molecule on human lymphocytes by special siRNA. AB - BACKGROUND: The B7/CD28 pathway provides critical costimulatory signals for complete T cell activation, and members of this pathway have served as useful targets for immunotherapeutic strategies. In this study, we investigated the RNA interference (RNAi) effect induced by small interfering RNA (siRNA) targeting CD28 mRNA on human lymphocytes and its specificity. METHODS: According to CD28 gene sequence, we designed and synthysized three different siRNAs (siRNA-1, siRNA 2, siRNA-3) containing 21 bases using Silencertrade mark siRNA construction kit. These siRNAs were transfected into freshly isolated human lymphocytes with Lipofectamine 2000 reagent. At 24-hour, 48-hour and 72-hour post transfection, these cells were collected and analyzed. The changes of surface expression of CD28 gene were detected by flow cytometry, and the changes of CD28 mRNA levels were determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). The cell viability of transfected lymphocytes was determined by methyl thiazolyl tetrazolium (MTT) assay and trypan blue dye exclusion assay. RESULTS: Three siRNAs (siRNA-1, siRNA-2, siRNA-3) specifically targeting CD28 mRNA were successfully designed and constructed. Flow cytometry analysis showed that a decrease in CD28 expression was detectable at 24-hour post transfection. Different siRNA showed different inhibition effects on CD28 expression. At 48 hour post transfection, the degrees of reduction with siRNA-1, siRNA-2 and siRNA 3 were 22.10% +/- 1.63%, 73.50% +/- 1.02% and 42.90% +/- 0.89% respectively compared with the control (P < 0.001). Neither of the groups transfected only with siRNA or lipo showed marked reduction in CD28 expression (3.15% +/- 0.75% and 4.55% +/- 0.80%) (P > 0.05). Moreover, lymphocytes treated with siRNA-co showed no marked reduction in CD28 expression (5.07% +/- 0.96%) (P > 0.05). The results of semi-quantitative RT-PCR assay indicated CD28 mRNA level was inhibited after transfection of specific siRNAs. At least 4-fold of reduction in siRNA-2 group occurred at 48-hour post transfection compared with the control (P < 0.001). MTT assay and trypan blue dye exclusion assay demonstrated that the viable cell rations of transfected lymphocytes were significantly reduced in siRNA-1, siRNA-2 and siRNA-3 groups at 48-hour post transfection (P < 0.01). The control groups showed no marked reduction in cell viability (P > 0.05). CONCLUSIONS: Three different siRNAs were synthesized and transfected into lymphocytes. They could reduce the expression of CD28 and the CD28 mRNA level. siRNA-2 was the most efficient. The cell viability reduced correspondingly. Therefore, the silencing effect on CD28 mRNA induced by siRNA may contribute to costimulatory blockade. This result show that siRNA may be useful for further study on graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo-BMT). PMID- 15788131 TI - Antiviral activity of cepharanthine against severe acute respiratory syndrome coronavirus in vitro. PMID- 15788132 TI - Relation of nm23 gene expression to CT sign and prognosis in peripheral nonsmall cell lung cancer. PMID- 15788133 TI - Positive association of RhoC gene overexpression with tumour invasion and lymphatic metastasis in gastric carcinoma. PMID- 15788134 TI - Protective effect of Shenfu injection on myocardial mitochondria injured by ischemia-reperfusion in rabbits. PMID- 15788135 TI - Transplantation of cholinergic neural stem cells in a mouse model of Alzheimer's disease. PMID- 15788136 TI - Clinical study on high-resolution CT and pulmonary function in severe acute respiratory syndrome patients during recovery phase. PMID- 15788137 TI - Parotid gland carcinosarcoma with follicular dendritic cell sarcoma as mesenchymal component: a case report. PMID- 15788138 TI - Imaging manifestations of von Hippel-Lindau disease: a report of 3 cases. PMID- 15788139 TI - Tentorial dural arteriovenous fistula successfully treated with surgical interruption of leptomeningeal venous drainage: a case report. PMID- 15788140 TI - Cure of an old pediatric femoral neck fracture: a case report. PMID- 15788141 TI - Atopy patch test reaction to airborne allergens in the diagnosis of atopic dermatitis. AB - The aim of the study was to evaluate the possible use of atopy patch test in the diagnosis of atopic dermatitis and to characterize an optimal standardized system for atopy patch test in terms of allergen concentrations and time of allergen exposure. The study included 36 patients with atopic dermatitis and IgE-mediated airborne allergy. Patients presented positive results of skin prick tests and serum antigen specific IgE against house dust mite allergens and/or selected grass pollen allergens. Control groups consisted either of patients with allergic rhinitis (control group 1) or healthy volunteers with no signs or symptoms of atopy (control group 2). Allergologic diagnostic workup consisted of skin prick test, serum antigen specific IgE and total IgE evaluation, atopy patch test with selected airborne allergens of different concentrations (0.1xSPT, 1xSPT and 10xSPT), time of allergen exposure (8, 24 and 48 h), and readings of the results (8, 24, 48 and 72 h). Positive results of atopy patch test with airborne allergens were obtained in 47.2% of atopic dermatitis patients and none of control subjects. Contact reaction itself and the intensity of reaction were demonstrated to correlate with allergen concentration and time of allergen exposure on atopy patch test. The dose and time response analysis showed the optimal concentration of allergens for atopy patch test to be 10xSPT, 500000 SBE/ml, and optimal evaluation time 24 and 48 h of allergen application. There was no correlation between atopy patch test results and mean serum concentrations of total or antigen specific IgE. Atopy patch test results did not correlate with localization of skin lesions, severity and extensiveness of skin inflammation. A significantly higher contact reactivity to airborne allergens was recorded in the group of atopic dermatitis patients with polyvalent allergy in comparison with atopic dermatitis patients allergic to only one aeroallergen. It is concluded that atopy patch test is the only provocation test currently available with clinical relevance for contact IgE-mediated sensitization in atopic dermatitis patients. Using petrolatum as a vehicle, allergen concentration of 500000 SBE/ml and evaluation time of 24 and 48 h of allergen application may lead to improved atopy patch test results. PMID- 15788142 TI - What do we really know about "tinea incognita"? AB - The term "tinea incognita" refers to diverse clinical presentation of mycotic infections modified by inappropriate use of topical or systemic corticosteroids. A 67-year-old male patient with a five-year history of generalized erythematous plaques on the trunk and extremities, previously treated with topical corticosteroids, is described. The lesions mainly showed a psoriasiform, some eczematous appearance, few of them showing a clinical picture of folliculitis. The native mycologic specimen was negative. The diagnosis was made on the basis of mycologic culture finding of Trichophyton interdigitale growth. Systemic and topical antimycotic therapy administered for two months resulted in complete regression of skin lesions. PMID- 15788143 TI - Classic Kaposi's sarcoma: a case report. AB - A 60-year-old male from the Mediterranean area presented with edematous right leg and livid nodules and macules on the skin of upper and lower extremities. Biopsy specimen obtained from the right upper leg showed a pathohistologic finding indicative of Kaposi's sarcoma. Polymerase chain reaction testing revealed HHV-8 in the skin lesion. Serology for HIV was negative. Additional examinations did not reveal dissemination of the disease. Negative HIV serology, normal laboratory findings and absence of immunosuppressant therapy in the patient's history confirmed the diagnosis of the classic form of Kaposi's sarcoma. PMID- 15788144 TI - A review of sunscreens and their adverse reactions. AB - Sunscreens are used to protect the skin from harmful effects of ultraviolet (UV) light but they do not completely prevent photocarcinogenesis, photoaging and photoimmunosuppression. They are useful for protection against UVB and short-wave UVA. Complete protection against long-wave UVA has not been achieved. There is no universally accepted method to evaluate UVA protection. Sun protection factor is a simple and internationally used method to compare sunscreen protection against UVB induced erythema. Adverse reactions to sunscreens are not common but they should be considered especially in persons with pre-existing eczematous conditions or photodermatoses. The use of sunscreens has increased steadily over the last decade; as a result, allergy and photoallergy to UV filters are now more frequent than in the past. Sensitization can occur from the various sunscreening agents and from the excipients included in formulations. An overview of sunscreens, their effectiveness, and adverse reactions is presented. PMID- 15788145 TI - Malignant melanoma--future prospects. AB - As the incidence and already high mortality rates of malignant melanoma have been steadily increasing in recent decades, the early detection and excision of malignant melanoma have imposed as the most important task. Staging of malignant melanoma is determined according to the level of invasion (Clark level) and vertical thickness (Breslow scale). Besides operative therapy, which is the only effective treatment for malignant melanoma, postoperative adjuvant chemotherapy, immunotherapy, radiotherapy, and biologic therapy also are of great importance. In recent years, immunologic strategies including tumor vaccine and adjuvant therapy with interferon-alfa have been attempted to improve survival of patients with more advanced malignant melanoma. A recent melanoma research has focused on target therapy such as immunotherapy (vaccines, monoclonal antibodies, dendritic cells) and gene therapy. Genetic immunization has become an attractive strategy for the development of melanoma vaccines, because a number of antigens recognized by cellular components of the immune system have been identified at the molecular level. Numerous chemotherapeutic agents have shown activity in the treatment of metastatic malignant melanoma, such as dacarbazine (dimethyl triazene imidazole carboxamide); other agents have been used, however, with less success. However, a very modest effect was recorded in advanced malignant melanoma. There are many experimental trials using combined therapy for malignant melanoma, including chemotherapy (dimethyl triazene imidazole carboxamide) and biologic therapy (interleukin (IL)-2, interferon (IFN)-gamma, IFN-alfa). The results obtained open particularly interesting prospects in the field of malignant melanoma with high relevance for its development and progression. Molecular therapeutics and vaccine development will probably be an important focus for the future melanoma treatment. PMID- 15788146 TI - Body dysmorphic disorder: more than meets the eye. AB - Body dysmorphic disorder is a relatively common yet long unrecognized problem affecting mainly adolescents and young adults of both sexes. It is an obsessive disease of such intensity that it interferes with daily activities. Patients typically focus on imagined major defects in the face, nose, skin or hair. Patients often seek multiple physician assessment, but body dysmorphic disorder may be disabling and even life-threatening as a result of depression and suicidal ideation. Treatment with serotonin reuptake inhibitors and cognitive behavioral therapy may be beneficial in many patients. PMID- 15788147 TI - The pathogenesis of acne. AB - Acne vulgaris is a multifactorial disease of as yet incompletely elucidated etiology and pathogenesis. The following have been identified as the most significant factors: follicular hyperkeratosis, increased sebum secretion, Propionibacterium (P.) acnes, and inflammation. Increased sebum production and follicular hyperkeratosis result in the development of microcomedones, and changes in follicular milieu in intensive growth of P. acnes. P. acnes secretes several proinflammatory products, which play an important role in the development of inflammation. These include lipases, proteases, hyaluronidases, and chemotactic factors. Immune response to P. acnes includes humoral and cell mediated immunity as well as complement activation. Recent results indicate that keratinocytes and sebocytes, as major components of pilosebaceous unit, may act as immune cells and may be activated by P. acnes via toll-like receptors (TLRs) and CD14, and through CD1 molecules may recognize altered lipid content in sebum, followed by the production of inflammatory cytokines. PMID- 15788148 TI - Etiopathogenesis of atopic dermatitis--an overview. AB - Atopic eczema/dermatitis syndrome is a term that covers different subtypes of atopic dermatitis. The "intrinsic" type of atopic dermatitis is non-IgE associated, and the "extrinsic" type is IgE-associated atopic eczema/dermatitis syndrome. In the etiopathogenesis of atopic dermatitis there are well known interactions among genetic, environmental, skin barrier, immune factors, and stress. Genetic factors determine the expression of atopic dermatitis as pure or mixed with concomitant respiratory or intestinal allergy, depending on genetic susceptibility. Immunologic abnormalities of type I and type IV reactions have been described in patients with atopic dermatitis. Immunologic triggers are aeroallergens, food allergens, microbial products, autoallergens and contact allergens. Immune reactions determine many features of atopic dermatitis. These immune reactions also include cell mediated or delayed hypersensitivity. The currently accepted model proposes a predominant Th2 cytokine milieu in the initiating stages of acute atopic dermatitis lesions, and a mixed Th1 and Th2 pattern in chronic lesions. A two-phase model includes Th2 initiation with attraction of macrophages and eosinophils, which in turn produce interleukin 12 that is the activator of Th1 type response. Atopic dermatitis skin contains an increased number of IgE-bearing Langerhans cells which bind allergens via the high-affinity IgE receptor (FcepsilonRI). Langerhans cells play an important role in cutaneous allergen presentation to Th2 cells via major histocompatibility molecules. Eosinophilia and IgE production are influenced by type 2 cytokines. Degranulation of eosinophils occurs in the dermis with the release of toxic proteins such as major basic protein and could account for much of the inflammation. Mast cells are increased in number and produce mediators other than histamine that induce pruritus and may have an effect on interferon gamma expression. Mast cells produce a number of proinflammatory cytokines. There is an elevated production of prostaglandin E2 by peripheral monocytes. Prostaglandin E2 has at least two potential roles in the initiation of atopic dermatitis. Firstly, it reduces interferon-gamma production by T helper cells, thereby favoring the initial, dominant Th2 immune response; and secondly, it directly enhances IgE production by B lymphocytes with an increased secretion of interleukin 4, interleukin 5 and interleukin 13. Many lesions of atopic dermatitis result from scratching, thus it is tempting to speculate that immune perturbations in genetically predisposed individuals provoke the release of local pruritogens and keratinocyte-derived cytokines, which then further exacerbate the previously described immune response. PMID- 15788149 TI - Pimecrolimus--a safe and effective local immunomodulator in the treatment of inflammatory skin diseases. AB - Pimecrolimus (SDZ ASM 981), an ascomycin derivative, as one of the new classes of immunomodulating macrolactams, is specifically effective in the treatment of inflammatory skin diseases. The interest in pimecrolimus is highly important for its significant anti-inflammatory activity, cell-selective inhibition of inflammatory cytokines, immunomodulatory capabilities, and low systemic immunosuppressive potential. The mechanism of action of pimecrolimus is the blockage of T cell activation, blocking signal transduction pathways in T cells, and inhibition of the synthesis of inflammatory cytokines, specifically Th1- and Th2-type cytokines. Several studies have evaluated the effectiveness of pimecrolimus as the treatment of choice for inflammatory skin diseases. PMID- 15788150 TI - Mitogenic and anti-proliferative activity of a lectin from the tubers of Voodoo lily (Sauromatum venosum). AB - A new lectin with the potent mitogenic and in vitro anti-proliferative activity was isolated from the tubers of a wild monocotyledonous plant Sauromatum venosum (Schott), from the family Araceae, by affinity chromatography on the asialofetuin linked amino-activated silica beads. The apparent native molecular mass of S. venosum lectin (SVL), as determined by gel filtration chromatography, was 54 kDa. In HPLC, size exclusion and cation exchange chromatography, SVL gave a single peak and also a single band of 13.5 kDa in SDS-PAGE, pH 8.3, under reducing and non-reducing conditions, indicating that the lectin is composed of four identical subunits. S. venosum lectin agglutinated rabbit, rat, sheep and guinea pig erythrocytes but reacted with goat erythrocytes after the neuraminidase treatment. However, SVL was unable to agglutinate human ABO blood group erythrocytes even after treatment with neuraminidase. SVL was inhibited by N acetyl-D-Lactosamine (LacNAc), which is an important marker in various carcinomas and a complex desialylated glycoprotein, asialofetuin. The amino acid composition showed that lectin contained a high amount of aspartic acid and glycine but totally devoid of cysteine. However, trace amounts of methionine was present. The lectin showed a potent mitogenic response towards BALB/c splenocytes and human lymphocytes. As the mitogenic stimulation was more than that of Con A, a standard well-known plant mitogen and the response of this lectin was almost double than that of Con A. This lectin is endowed with proliferation of T cells as revealed by IL-2 bioassay but showed no production of immunoglobulins thus indicating the non-stimulation of B cells. SVL significantly inhibited the proliferation of murine cancer cell-lines, i.e., WEHI-279 to 84.6%, J774 to 81%, P388D1 to 74% and A-20 to 47%. In addition, the in vitro anti-proliferative activity of SVL was also evaluated against nine human cancer cell lines representing different organs and tissues namely, T-47D (breast), SiHa (cervix), SK-N-MC (CNS), SK-N-SH (CNS), SW-620 (colon), HT-29 (colon), HEP-2 (liver), OVCAR-5 (ovary) and PC-3 (prostate). SVL showed a significant inhibition towards the entire cell lines except the cell lines from CNS, which showed partial response in comparison to a standard anticancer drug adriamycin which was used at a concentration of 5 x 10( 5) M. Thus the anti-proliferative ability of SVL may be helpful in identification of new lectin probes that can lead to better understanding in the detection and study of certain types of cancer. PMID- 15788151 TI - Development of a spot reliability evaluation score for DNA microarrays. AB - We developed a reliability index named SRED (Spot Reliability Evaluation Score for DNA microarrays) that represents the probability that the calibrated gene expression level from a DNA microarray would be less than a factor of 2 different from that of quantitative real-time polymerase chain reaction assays whose dynamic quantification range is treated statistically to be similar to that of the DNA microarray. To define the SRED score, two parameters, the reproducibility of measurement value and the relative expression value were selected from nine candidate parameters. The SRED score supplies the probability that the expression level in each spot of a microarray is less than a certain-fold different compared to other expression profiling data, such as QRT-PCR. This score was applied to approximately 1,500,000 points of the expression profile in the RIKEN Expression Array Database. PMID- 15788152 TI - The first complete chloroplast genome sequence of a lycophyte, Huperzia lucidula (Lycopodiaceae). AB - We used a unique combination of techniques to sequence the first complete chloroplast genome of a lycophyte, Huperzia lucidula. This plant belongs to a significant clade hypothesized to represent the sister group to all other vascular plants. We used fluorescence-activated cell sorting (FACS) to isolate the organelles, rolling circle amplification (RCA) to amplify the genome, and shotgun sequencing to 8x depth coverage to obtain the complete chloroplast genome sequence. The genome is 154,373 bp, containing inverted repeats of 15,314 bp each, a large single-copy region of 104,088 bp, and a small single-copy region of 19,657 bp. Gene order is more similar to those of mosses, liverworts, and hornworts than to gene order for other vascular plants. For example, the Huperzia chloroplast genome possesses the bryophyte gene order for a previously characterized 30 kb inversion, thus supporting the hypothesis that lycophytes are sister to all other extant vascular plants. The lycophyte chloroplast genome data also enable a better reconstruction of the basal tracheophyte genome, which is useful for inferring relationships among bryophyte lineages. Several unique characters are observed in Huperzia, such as movement of the gene ndhF from the small single copy region into the inverted repeat. We present several analyses of evolutionary relationships among land plants by using nucleotide data, inferred amino acid sequences, and by comparing gene arrangements from chloroplast genomes. The results, while still tentative pending the large number of chloroplast genomes from other key lineages that are soon to be sequenced, are intriguing in themselves, and contribute to a growing comparative database of genomic and morphological data across the green plants. PMID- 15788153 TI - Testosterone responsiveness of spleen and liver in female lymphotoxin beta receptor-deficient mice resistant to blood-stage malaria. AB - Disrupted signaling through lymphotoxin beta receptor (LTbetaR) results in severe defects of the spleen and even loss of all other secondary lymphoid tissues, making mice susceptible to diverse infectious agents. Surprisingly, however, we find that female LTbetaR-deficient mice are even more resistant to blood stages of Plasmodium chabaudi malaria than wild-type C57BL/6 mice. Higher resistance of LTbetaR-deficient mice correlates with an earlier onset of reticulocytosis, and the period of anemia is shorter. After surviving fulminant parasitemias of about 35%, mice develop long-lasting protective immunity against homologous rechallenge, with both spleen and liver acting as anti-malaria effectors. Testosterone suppresses resistance, i.e. all mice succumb to infections during or shortly after peak parasitemia. At peak parasitemia, testosterone does not essentially affect cellularity and apoptosis in the spleen, but aggravates liver pathology in terms of increased cell swelling, numbers of apoptotic and binucleated cells and reduced serum alkaline phosphatase levels, and conversely, reduces inflammatory lymphocytic infiltrates in the liver. In the spleen, hybridization of cDNA arrays identified only a few testosterone-induced changes in gene expression, in particular upregulation of INFgamma and IFN-regulated genes. By contrast, a much larger number of testosterone-affectable genes was observed in the liver, including genes involved in regulation of the extracellular matrix, in chemokine and cytokine signaling, and in cell cycle control. Collectively, our data suggest that testosterone dysregulates the inflammatory response in spleen and liver during their differentiation to anti malaria effectors in malaria-resistant female LTbetaR-deficient mice, thus contributing to the testosterone-induced lethal outcome of malaria. PMID- 15788154 TI - Strain-specific expression profiles of virulence genes in Helicobacter pylori during infection of gastric epithelial cells and granulocytes. AB - Helicobacter pylori expresses a variety of known virulence-associated factors, whose expression is likely to be dependent on the ecological niche of this pathogen. Here, we compared the temporal changes in the level of virulence associated gene transcription in H. pylori strains isolated from patients with different pathology. Our aim was to study the coordinated gene expression profiles of these virulence factors during infection of AGS gastric epithelial cells and granulocytes. Using real-time quantitative (TaqMan) RT-PCR, we determined the mRNA expression of cagA, ureA, napA, katA, vacAs1 and vacAs2 alleles in a time course up to 6 h. The expression profiles of the investigated genes vary according to the strain, and were mainly either upregulated or unchanged upon bacterial contact with AGS cells. In contrast, upon contact with granulocytes, the majority of the genes were repressed in H. pylori. The following major results were obtained: (i) genetically diverse H. pylori exhibit different mRNA expression profiles, (ii) the expression patterns were strain specific and time-dependent and (iii) the regulation of expression profiles was host cell dependent. These data were statistically significant and suggest that contact with target cells leads to an active cross-talk between the pathogen and its host. The use of Taqman-PCR to analyse the expression of mRNA of a bacterial pathogen in response to a changing host environment enabled us to identify variable and strain-specific transcription profiles in a sensitive and reproducible manner. PMID- 15788155 TI - The contribution of reactive nitrogen and oxygen species to the killing of Francisella tularensis LVS by murine macrophages. AB - Intracellular killing of Francisella tularensis by macrophages depends on interferon-gamma (IFN-gamma)-induced activation of the cells. The importance of inducible nitric oxide synthase (iNOS) or NADPH phagocyte oxidase (phox) for the cidal activity was studied. Murine IFN-gamma-activated peritoneal exudate cells (PEC) produced nitric oxide (NO), measured as nitrite plus nitrate, and superoxide. When PEC were infected with the live vaccine strain, LVS, of F. tularensis, the number of viable bacteria was at least 1000-fold lower in the presence than in the absence of IFN-gamma after 48 h of incubation. PEC from iNOS gene-deficient (iNOS-/-) mice killed F. tularensis LVS less effectively than did PEC from wild-type mice. PEC from phox gene-deficient (p47phox-/-) mice were capable of killing the bacteria, but killing was less efficient, although still significant, in the presence of NG-monomethyl-L-arginine (NMMLA), an inhibitor of iNOS. A decomposition catalyst of ONOO-, FeTPPS, completely reversed the IFN gamma-induced killing of F. tularensis LVS. Under host cell-free conditions, F. tularensis LVS was exposed to S-nitroso-acetyl-penicillamine (SNAP), which generates NO, or 3-morpholinosydnonimine hydrochloride (SIN-1), which generates NO and superoxide, leading to formation of ONOO-. During 6 h of incubation, SNAP caused no killing of F. tularensis LVS, whereas effective killing occurred in the presence of equimolar concentrations of SIN-1. The results suggest that mechanisms dependent on iNOS and to a minor degree, phox, contribute to the IFN gamma-induced macrophage killing of F. tularensis LVS. ONOO- is likely to be a major mediator of the killing. PMID- 15788157 TI - Can we talk to robots? Ten-month-old infants expected interactive humanoid robots to be talked to by persons. AB - As technology advances, many human-like robots are being developed. Although these humanoid robots should be classified as objects, they share many properties with human beings. This raises the question of how infants classify them. Based on the looking-time paradigm used by [Legerstee, M., Barna, J., & DiAdamo, C., (2000). Precursors to the development of intention at 6 months: understanding people and their actions. Developmental Psychology, 36, 5, 627-634.], we investigated whether 10-month-old infants expected people to talk to a humanoid robot. In a familiarization period, each infant observed an actor and an interactive robot behaving like a human, a non-interactive robot remaining stationary, and a non-interactive robot behaving like a human. In subsequent test trials, the infants were shown another actor talking to the robot and to the actor. We found that infants who had previously observed the interactive robot showed no difference in looking-time between the two types of test events. Infants in the other conditions, however, looked longer at the test event where the second experimenter talked to the robot rather than where the second experimenter talked to the person. These results suggest that infants interpret the interactive robot as a communicative agent and the non-interactive robot as an object. Our findings imply that infants categorize interactive humanoid robots as a kind of human being. PMID- 15788158 TI - Consistent (but not variable) names as invitations to form object categories: new evidence from 12-month-old infants. AB - Recent research documents that for infants just beginning to produce words on their own, novel words highlight commonalities among named objects and, in this way, serve as invitations to form categories. The current experiment identifies more precisely the source of this invitation. We asked whether applying a consistent name to a set of distinct objects is crucial to categorization, or whether variable names might serve the same conceptual function. The evidence suggests that for 12-month-old infants, consistency in naming is critical. Infants hearing a single consistent novel noun for a set of distinct objects successfully formed object categories. Infants hearing different novel nouns for the same set of objects did not. These results lend strength and greater precision to the argument that naming has powerful and rather nuanced conceptual consequences for infants as well as for mature speakers. PMID- 15788159 TI - Visual arguments. AB - Three experiments investigated the use of verb argument structure by tracking participants' eye movements across a set of related pictures as they listened to sentences. The assumption was that listeners would naturally look at relevant pictures as they were mentioned or implied. The primary hypothesis was that a verb would implicitly introduce relevant entities (linguistic arguments) that had not yet been mentioned, and thus a picture corresponding to such an entity would draw anticipatory looks. For example, upon hearing ...mother suggested..., participants would look at a potential recipient of the suggestion. The only explicit task was responding to comprehension questions. Experiments 1 and 2 manipulated both the argument structure of the verb and the typicality/co occurrence frequency of the target argument/adjunct, in order to distinguish between anticipatory looks to arguments specifically and anticipatory looks to pictures that were strongly associated with the verb, but did not have the linguistic status of argument. Experiment 3 manipulated argument status alone. In Experiments 1 and 3, there were more anticipatory looks to potential arguments than to potential adjuncts, beginning about 500 ms after the acoustic onset of the verb. Experiment 2 revealed a main effect of typicality. These findings indicate that both real world knowledge and argument structure guide visual attention within this paradigm, but that argument structure has a privileged status in focusing listener attention on relevant aspects of a visual scene. PMID- 15788160 TI - The influence of contextual contrast on syntactic processing: evidence for strong interaction in sentence comprehension. AB - The present study compares the processing of unambiguous restrictive and non restrictive relative clauses (RCs) within both a null context and a supportive discourse using a self-paced reading methodology. Individuals read restrictive RCs more slowly than non-restrictive RCs in a null context, but processed restrictive RCs faster than non-restrictive RCs in supportive context, resulting in an interaction between context and RC type. These results provide evidence for two theoretical points. First, principles analogous to those in referential theory [Altmann G. T. M., & Steedman, M. (1988). Interaction with context during human sentence processing. Cognition, 30, 191-238; Crain, S., & Steedman, M. (1985). On not being led up the garden path: The use of context by the psychological parser. In D. Dowty, L. Karttunnen, A. Zwicky (Eds.), Natural language parsing. Cambridge, UK: Cambridge University Press] apply not only in resolving ambiguity but also in processing unambiguous sentences. Second, the discourse context can guide and facilitate interpretive processing. This result suggests that intrasentential factors such as syntax are not autonomous from contextual processing, contrary to the modularity hypothesis [Fodor, J. A. (1983). Modularity of mind. Cambridge, MA: MIT Press]. PMID- 15788156 TI - A double-dissociation in infants' representations of object arrays. AB - Previous studies show that infants can compute either the total continuous extent (e.g. Clearfield, M.W., & Mix, K.S. (1999). Number versus contour length in infants' discrimination of small visual sets. Psychological Science, 10(5), 408 411; Feigenson, L., & Carey, S. (2003). Tracking individuals via object-files: evidence from infants' manual search. Developmental Science, 6, 568-584) or the numerosity (Feigenson, L., & Carey, S. (2003). Tracking individuals via object files: evidence from infants' manual search. Developmental Science, 6, 568-584) of small object arrays. The present experiments asked whether infants can compute both extent and number over a given array. Experiment 1 used a habituation procedure to show that 7-month-old infants can compute numerosity when the objects in the array contrast in color, pattern, and texture. Experiment 2 revealed that, with these heterogeneous arrays, infants no longer represent the array's total continuous extent. Since previous work shows that infants compute continuous extent but not numerosity when objects have identical rather than contrasting properties, these results form a double dissociation. Infants computed number but not extent over representations of contrasting objects, and computed extent but not number over representations of identical objects. PMID- 15788162 TI - Interference effects demonstrate distinct roles for visual and motor imagery during the mental representation of human action. AB - Four experiments were completed to characterize the utilization of visual imagery and motor imagery during the mental representation of human action. In Experiment 1, movement time functions for a motor imagery human locomotion task conformed to a speed-accuracy trade-off similar to Fitts' Law, whereas those for a visual imagery object motion task did not. However, modality-specific interference effects in Experiment 2 demonstrate visual and motor imagery as cooperative processes when the action represented is tied to visual coordinates in space. Biomechanic-specific motor interference effects found in Experiment 3 suggest one basis for separation of processing channels within motor imagery. Finally, in Experiment 4 representations of motor actions were found to be generated using only visual imagery under certain circumstances: namely, when the imaginer represented the motor action of another individual while placed at an opposing viewpoint. These results suggest that the modality of representation recruited to generate images of human action is dependent on the dynamic relationship between the individual, movement, and environment. PMID- 15788163 TI - Inhibitory activity of polyhydroxycarboxylate chelators against recombinant NF kappaB p50 protein-DNA binding. AB - The inhibitory effect of 7,8-dihydroxy-4-methylcoumarin (7,8-DHMC), 5,7-dihydroxy 4-methylcoumarin (5,7-DHMC), and gallic acid on the DNA binding of recombinant p50 protein and their interaction with zinc ion were studied. Electrophoretic mobility shift assay (EMSA) using p50 and biotin labeled DNA has shown that gallic acid is more effective than the dihydroxycoumarins in inhibiting the p50 DNA binding. Molecular modeling studies suggest an explanation for these observations. Effect of the addition of zinc after p50-DNA-binding inhibition by gallic acid was also studied. Chemical speciation and formation constant studies show that gallic acid forms a more stable 1:1 complex with zinc ion in comparison to the dihydroxycoumarins. PMID- 15788161 TI - When the ordinary seems unexpected: evidence for incremental physical knowledge in young infants. AB - According to a recent account of infants' acquisition of their physical knowledge, the incremental-knowledge account, infants form distinct event categories, such as occlusion, containment, support, and collision events. In each category, infants identify one or more vectors which correspond to distinct problems that must be solved. For each vector, infants acquire a sequence of variables that enables them to predict outcomes within the vector more and more accurately over time. This account predicts that infants who have acquired only a few of the variables in a sequence should err in two ways in violation-of expectation tasks: (1) they should view impossible events consistent with their incomplete knowledge as expected (errors of omission), and (2) they should view possible events inconsistent with their incomplete knowledge as unexpected (errors of commission). Many reports have shown that infants who have not yet identified a variable in an event category produce errors of omission: they fail to view impossible events involving the variable as unexpected. However, there has been no report revealing errors of commission in infants' responses to possible events. The present research examined whether 3- and 2.5-month-old infants, whose knowledge of occlusion events is very limited, would produce errors of commission as well as errors of omission when responding to these events. At 3 months of age, infants viewed as unexpected a possible event in which a tall cylinder became visible when passing behind a tall screen with a very large opening extending from its upper edge. At 2.5 months, infants viewed as unexpected a possible event in which a tall cylinder became visible when passing behind a tall screen with a very large opening extending from its lower edge. These findings provide a new kind of evidence for the incremental-knowledge account, and more generally for the notion that infants, like older children and adults, engage in rule-based reasoning about physical events. PMID- 15788164 TI - Stereospecificity of hydride transfer in NAD+-catalyzed 2-deoxy-scyllo-inosose synthase, the key enzyme in the biosynthesis of 2-deoxystreptamine-containing aminocyclitol antibiotics. AB - The key enzyme in the biosynthesis of clinically important aminocyclitol antibiotics is 2-deoxy-scyllo-inosose synthase (DOIS), which converts ubiquitous d-glucose 6-phosphate (G-6-P) into the specific carbocycle, 2-deoxy-scyllo inosose with an aid of NAD(+)-NADH recycling. The NAD(+)-dependent first step of the DOIS reaction was examined in detail by the use of 6-phosphonate and 6 homophosphonate analogs of G-6-P. Both analogs showed competitive inhibition against the DOIS reaction with K(i) values of 1.3 and 2.8 mM, respectively, due to their inability for the subsequent phosphate elimination. Based on the direct spectrophotometric observation of NADH formed by the hydride transfer from 6 phosphonate to NAD(+), the stereospecificity of the hydride transfer in the DOIS reaction was analyzed with 6-[4-(2)H]phosphonate and was found to be pro-R specific. PMID- 15788166 TI - A theoretical study of the different radical-scavenging activities of catechin, quercetin, and a rationally designed planar catechin. AB - To improve the radical-scavenging activity of catechin, a planar catechin analogue was designed and synthesized by Fukuhara [J. Am. Chem. Soc. 124 (2002) 5952]. Although the planar catechin is less active than quercetin, it is much more active than catechin in its ability to scavenge galvinoxyl radical, suggesting that the rational design was successful. However, an interesting question remains: what is the basis for the enhanced radical-scavenging activity of the planar catechin? By DFT calculations, we determined that the galvinoxyl radical is scavenged through an electron-transfer mechanism rather than a hydrogen-atom-transfer mechanism. Moreover, the antioxidant anion, derived from proton dissociation, plays a key role in the radical-scavenging process. Hence, the different radical-scavenging activities of the three antioxidants may result from the different ionization potentials of their anions. PMID- 15788167 TI - Total synthesis of 12,13-desoxyepothilone B (Epothilone D). AB - A highly convergent total synthesis of 12,13-desoxyepothilone B (4, Epothilone D) is described involving the coupling of vinyl iodide (5) and olefin (6). Key steps in the synthesis are the introduction of chirality at C15 via highly enantioselective lipase-mediated enzymatic resolution, diastereoselective alkylation at C8, highly diastereoselective Evans aldol reaction to establish C6 C7, and Mukaiyama aldol reaction to introduce chiral center C3. Palladium catalyzed Suzuki coupling of (5) and (6) provided the methyl ester (27), which was converted to 12,13-desoxyepothilone B (4). PMID- 15788165 TI - The importance of the amide bond nearest the thiol group in enzymatic reactions of coenzyme A. AB - Analogues of coenzyme A (CoA) and of CoA thioesters have been prepared in which the amide bond nearest the thiol group has been modified. An analogue of acetyl CoA in which this amide bond is replaced with an ester linkage was a good substrate for the enzymes carnitine acetyltransferase, chloramphenicol acetyltransferase, and citrate synthase, with K(m) values 2- to 8-fold higher than those of acetyl-CoA and V(max) values from 14 to >80% those of the natural substrate. An analogue in which an extra methylene group was inserted between the amide bond and the thiol group showed less than 4-fold diminished binding to the three enzymes but exhibited less than 1% activity relative to acetyl-CoA with carnitine acetyltransferase and no measurable activity with the other two enzymes. Analogues of several CoA thioesters in which the amide bond was replaced with a hemithioacetal linkage exhibited no measurable activity with the appropriate enzymes. The results indicate that some aspects of the amide bond and proper distance between this amide and the thiol/thioester moiety are critical for activity of CoA ester-utilizing enzymes. PMID- 15788168 TI - Effects of sulfonamide and tetracycline antibiotics on soil microbial activity and microbial biomass. AB - Increasingly often soil residual concentrations of pharmaceutical antibiotics are detected, while their ecotoxic relevance is scarcely known. Thus, dose related effects of two antibiotics, sulfapyridine and oxytetracycline, on microorganisms of two different topsoils were investigated. The fumigation-extracted microbial C (E(C)) and ergosterol were determined to indicate soil microbial and fungal biomass, respectively. Microbial activity was tested as basal respiration (BR), dehydrogenase activity (DHA), substrate-induced respiration (SIR), and Fe(III) reduction. The BR and DHA were uninfluenced even at antibiotic concentrations of 1000 microg g(-1). This revealed that an activation of microbial growth through nutrient substrate addition is required to test possible effects of the bacteriostatic antibiotics. In addition, the effects of both antibiotics were time dependent, showing that short-term tests were not suitable. Clear dose response relations were determined with SIR when the short-term incubation of 4h was extended into the growth phase of the microorganisms (24 and 48 h). The Fe(III) reduction test, with a 7-d incubation, was also found to be suitable for toxicity testing of antibiotics in soils. Effective doses inhibiting the microbial activity by 10% (ED(10)) ranged from total antibiotic concentrations of 0.003-7.35 microg g(-1), depending on the antibiotic compound and its soil adsorption. Effective solution concentrations (EC(10)), calculated from distribution coefficients, ranged from 0.2 to 160 ng g(-1). The antibiotics significantly (p<0.05) reduced numbers of soil bacteria, resulting in dose related shifts in the fungal:bacterial ratio, which increased during 14 d, as determined from analysis of ergosterol and E(C). It was concluded that pharmaceutical antibiotics can exert a temporary selective pressure on soil microorganisms even at environmentally relevant concentrations. PMID- 15788169 TI - Quantitative structure-activity relationships of nitroaromatics toxicity to the algae (Scenedesmus obliguus). AB - The DFT-B3LYP method, with the basis set 6-311G( * *), was employed to calculate the molecular geometries and electronic structures of 25 nitroaromatics. The acute toxicity (-lgEC(50)) of these compounds to the algae (Scenedesmus obliguus) along with hydrophobicity described by logK(OW), and two quantum chemical parameters-energy of the lowest unoccupied molecular orbital, E(LUMO), and the charge of the nitro group, [ForQ(NO2), were used to establish the quantitative structure-activity relationships (QSARs). For 18 mononitro derivatives, the hydrophobicity parameter logK(OW) could interpret the toxic mechanism successfully. Dinitro aromatic compounds were susceptible to be reduced to aniline for their electrophilic nature. Their toxicity was controlled mainly by electronic factors instead of hydrophobicity. The electronic parameters, E(LUMO) and Q(NO2), were used to yield the following model: -lg EC(50) = 3.746 - 25.053 E(LUMO) + 6.481 Q(NO2) (n=22, R=0.926, SE=0.206, F=56.854, P<0.001). The predicted toxic values using the above equation are in good agreement with the experimental values. PMID- 15788170 TI - European union system for the evaluation of substances: the second version. AB - This publication presents major changes in the assessment of the risks of chemicals to human health and the environment as implemented in the second version of the European Union System for the Evaluation of Substances, EUSES 2.0. EUSES is a harmonised quantitative risk assessment tool for chemicals. It is the PC-implementation of the technical guidelines developed within the framework of EU chemical legislation for industrial chemicals and biocides. As such, it is designed to support decision making by risk managers in government and industry and to assist scientific institutions in the risk assessment for these substances. The development of EUSES 2.0 is a co-ordinated project of the European Chemicals Bureau, EU Member States and the European chemical industry. Several model concepts, the technical background and the user interface of EUSES have been improved considerably. Major changes in the environmental assessment such as the implementation of emission scenario documents for industrial chemicals and biocides, the addition of the marine risk assessment, the enhancement of the regional model to include global scales, and improvements in the secondary poisoning and environmental effects modelling will be discussed. The update of the human risk assessment module in EUSES focuses on the risk characterisation for both threshold and non-threshold substances with, among others, the introduction of assessment factors. The performance of EUSES is illustrated in an example showing the human and environmental risk assessment of a sanitation disinfectant for private use. PMID- 15788172 TI - Uptake and enantioselective elimination of chlordane compounds by common carp (Cyprinus carpio, L.). AB - An analytical method involving supercritical fluid extraction (SFE) followed by a two-dimensional gas chromatography (2D-GC) analysis was developed to determine the concentration (first GC) and enantiomeric ratio (second GC) of cis- and trans chlordanes at the ppb (ng/g) level in fish tissue. The SFE method allowed concentration of the compounds of interest, and reduced the number of extraction and sample clean-up manipulations as compared to classical solvent extraction techniques. Four hundred common carp fingerling (Cyprinus carpio, L.) were exposed for three days to water containing 5 ppb (5 ng/g) technical grade chlordane containing about 1 ppb of chlordane isomers. The fish concentrated the pesticides more than 200 times (162 and 312 ng/g of cis- and trans-chlordane, respectively). However, the uptake is not enantioselective. The concentration of the principle constituents and their enantiomeric ratio was followed during a fifty days growth period in chlordane free water. The first order decay of concentration was observed with a half time of about 18 days for both the cis- and trans-chlordane isomers. However it was found that the enantiomeric ratio of the trans-chlordane was significantly altered during this short period of time, decreasing from ER=1 to ER=0.7, while no enantiomeric changes were observed for the cis-chlordane. It seems that the (-)-trans-chlordane is metabolized significantly faster (t(1/2-)=15 days) in the river carp fish than the (+)-trans enantiomer (t(1/2+)=20 days). PMID- 15788171 TI - Blood concentrations of polycyclic musks in healthy young adults. AB - Knowledge on the concentration of polycyclic musk fragrance compounds in human blood is sparse. This study examined the concentrations of six polycyclic musks in blood samples from healthy volunteers. Blood was taken from hundred healthy students of the Medical University of Vienna. The lipophilic fraction was extracted and after purification analyzed by GC-MS. Study participants also completed a questionnaire on the use of cosmetics, about nutrition and other life style aspects. Two compounds -- galaxolide and tonalide -- were identified in higher percentages of the blood plasma samples. Maximum plasma levels over 100 ng/l were also only found for galaxolide (4100 ng/l) and tonalide (800 ng/l). Women showed significantly higher levels than men. In a statistical multivariate approach only use of body lotion and age were predictive of positive galaxolide concentrations. For tonalide no significant predictor could be found. The findings mirror the replacement of nitro musk fragrances by polycyclic musks, mainly galaxolide. The high concentrations of galaxolide in human blood raise concern since few toxicological data are available. PMID- 15788173 TI - Avoidance behaviour of Enchytraeus albidus: effects of benomyl, carbendazim, phenmedipham and different soil types. AB - Enchytraeids are typical inhabitants of many soils, contributing to vital processes of this environmental compartment. Indirectly they are involved in regulating the degradation of organic matter, as well as improving the pore structure of the soil. Due to their behaviour, they are able to avoid unfavourable environmental conditions. Avoidance tests with enchytraeids, initially developed with earthworms by several authors, are quick and easy to perform. With these tests a first assessment of the toxicity of a (contaminated or spiked) soil is possible in just 48 h by using the reaction of the enchytraeids as measurement endpoint. In this period of time the organisms can choose between the control soil and the other soil (a contaminated or spiked or another soil with different physico-chemical properties). In the tests reported here, the enchytraeids were exposed to control soils spiked with the fungicides Benomyl and Carbendazim and the herbicide Phenmedipham. Several chemical concentrations were tested in order to evaluate the avoidance behaviour to toxic substances. In fact, often these short-term screening tests gave results showing avoidance at concentrations in a range similar to the acute test results but, higher than in chronic tests. Further tests are needed to decide whether the results gained in this study can be extrapolated to other chemicals. It is proposed to standardize the Enchytraeid Avoidance Test as it is currently done for the Earthworm Avoidance Test by the International Standard Organization (ISO). PMID- 15788174 TI - An integrated assessment of estrogenic contamination and biological effects in the aquatic environment of The Netherlands. AB - An extensive study was carried out in the Netherlands on the occurrence of a number of estrogenic compounds in surface water, sediment, biota, wastewater, rainwater and on the associated effects in fish. Compounds investigated included natural and synthetic hormones, phthalates, alkylphenol(ethoxylate)s and bisphenol-A. The results showed that almost all selected (xeno-)estrogens were present at low concentrations in the aquatic environment. Locally, they were found at higher levels. Hormones and nonylphenol(ethoxylate)s were present in concentrations that are reportedly high enough to cause estrogenic effects in fish. Field surveys did not disclose significant estrogenic effects in male flounder (Platichthys flesus) in the open sea and in Dutch estuaries. Minor to moderate estrogenic effects were observed in bream (Abramis brama) in major inland surface waters such as lowland rivers and a harbor area. The prevalence of feminizing effects in male fish is largest in small regional surface waters that are strongly influenced by sources of potential hormone-disrupting compounds. High concentrations of plasma vitellogenin and an increased prevalence of ovotestes occurred in wild male bream in a small river receiving a considerable load of effluent from a large sewage treatment plant. After employing in vitro and in vivo bioassays, both in situ and in the laboratory, we conclude that in this case hormones (especially 17 alpha-ethynylestradiol) and possibly also nonylphenol(ethoxylate)s are primarily responsible for these effects. PMID- 15788175 TI - Reproduction of the estuarine and marine amphipod Corophium volutator (Pallas) in laboratory for toxicity testing. AB - The acute whole sediment bioassay with the estuarine and marine amphipod Corophium volutator (Pallas) is broadly used within Europe. Hitherto, the test is carried out with field-collected animals. In order to provide a more standardized and continuously available test organism reproduction and growth experiments were performed for a period of one year in laboratory under simulated summer conditions (light/dark 16:8 at 15, 19, and 23 degrees C). C. volutator was the first time reproduced successfully in laboratory for several generations and independent from its natural life cycle also in winter. The females produced two to three broods with a mean number of 96 offspring. A mean growth rate of 0.07 mm per day was determined at 15 degrees C. The reproduction and growth experiments provide consequently the essential base for the development of a chronic toxicity test with C. volutator. PMID- 15788176 TI - PCB contamination from polysulphide sealants in residential areas-exposure and risk assessment. AB - From the late 1950s to the early 1970s elastic polysulphide sealants were used in outdoor seams between concrete blocks in prefabricated buildings. The sealants contained 5-30% polychlorinated biphenyls (PCBs). Due to the weathering of sealants in general and the replacement of seams with new PCB-free materials in the 1990s, PCBs have drifted to the soil adjacent to buildings. The objectives of this study were to survey PCB contamination in the surroundings of former PCB containing buildings and to evaluate the risks to human health. Samples from soil, and also from blood serum of residents, were collected to obtain data for exposure assessment. The health risk assessment was based on deterministic and probabilistic calculations for cancer and non-cancer risks. Soil ingestion and dermal contact were considered the main routes of exposure and children the most important exposed group. The mean total PCB concentration was 6.83 mg/kg within 2m of the buildings and 0.52 mg/kg within 3-10 m from of the buildings. The deterministic risk assessment with conservative parameters resulted in lifetime cancer risk estimates on the order of 10(-6)-10(-7). The lifetime average daily dose (LADD) for PCBs was less than 10% of the reference dose (RfD) 0.02 microg/kg day, which is based on immunosupression in monkeys. The LADD corresponding to the total site attributable exposure was less than 10% of the estimated average dietary PCB intake in Finland. Children can, however, in worst cases be exposed to daily doses near the level of the RfD. Low cost measures are recommended to reduce possible exposure of children. PMID- 15788177 TI - Influence of sediment on the fate and toxicity of a polyethoxylated tallowamine surfactant system (MON 0818) in aquatic microcosms. AB - The fate and toxicity of a polyethoxylated tallowamine (POEA) surfactant system, MON 0818, was evaluated in water-sediment microcosms during a 4-d laboratory study. A surfactant solution of 8 mg l(-1) nominal concentration was added to each of nine 72-l aquaria with or without a 3-cm layer of one of two natural sediments (total organic carbon (TOC) 1.5% or 3.0%). Control well water was added to each of nine additional 72-l aquaria with or without sediment. Water samples were collected from the microcosms after 2, 6, 24, 48, 72, and 96 h of aging to conduct 48-h toxicity tests with Daphnia magna and to determine surfactant concentrations. Elevated mortality of D. magna (43-83%) was observed in overlying water sampled from water-only microcosms throughout the 96-h aging period, whereas elevated mortality (23-97%) was only observed in overlying water sampled from water-sediment microcosms during the first 24h of aging. Measured concentrations of MON 0818 in water-only microcosms remained relatively constant (4-6 mg l(-1)) during the 96-h period, whereas the concentrations in overlying water from microcosms containing either of the two types of sediment dissipated rapidly, with half-lives of 13 h in the 3.0% TOC sediment and 18 h in the 1.5% TOC sediment. Both toxicity and the concentration of MON 0818 in overlying water decreased more rapidly in microcosms containing sediment with the higher percent TOC and clay and with a higher microbial biomass. Mortality of D. magna was significantly correlated with surfactant concentrations in the overlying water. These results indicate that the toxicity of the POEA surfactant in water rapidly declines in the presence of sediment due to a reduction in the surfactant concentration in the overlying water above the sediment. PMID- 15788178 TI - Comparative study on the environmental risk induced by several pyrethroids in estuarine and freshwater invertebrate organisms. AB - The acute toxicity of permethrin, resmethrin, and cypermethrin to four species of aquatic non-target invertebrate organisms, found in estuarine and freshwater ecosystems, was evaluated. Artemia franciscana and Brachionus plicatilis larvae, as estuarine organisms, and Brachionus calyciflorus and Thamnocephalus platyurus larvae, as freshwater organisms, were exposed for 24 h to concentrations of these pyrethroids, and the LC(50) values were compared. The freshwater organisms were more sensitive to these pyrethroids than estuarine organisms tested. A. franciscana larvae were more tolerant organisms than B. plicatilis larvae. The freshwater organisms tested have demonstrated to be a good alternative to the standard acute toxicity assays using Daphnia, although Brachionus plycatilis larvae were more sensitive to these pyrethroid insecticides than T. platyurus. Analysis of 24 h LC(50) values of these pyrethroids, determined by static bioassays, revealed that the rank order of toxicity was: permethrinCu>Pb>Cd and at pH 7 Cd>Zn, with precipitation affecting Cu and Pb behaviour. During the column leaching experiment, most of the heavy metals were irreversibly bound to the soil, but in the case of Cr some movement from soil into solution was observed. The results also showed that Cd, Cu, Pb and Zn were removed from the solution and adsorbed on the soil. No significant difference in the metal removal from single- and multi element solutions was observed. Overall, the urban residue behaved in a similar manner to mineral soils despite a significant component of anthropogenic solid materials. PMID- 15788193 TI - Evaluation of the concentration of HCH, DDT, HCB, PCB and PAH in the sediments along the lower stretch of Hugli estuary, West Bengal, northeast India. AB - Detailed analyses of persistent organic pollutants (POPs) such as hexachlorocyclohexane (HCH) isomers (HCHs), dichlorodiphenyltrichloro ethane (DDT) and its metabolites (DDTs), hexachlorobenzene (HCB) and congeners of polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) in surface sediments from the coastal estuarine environment of West Bengal, northeast India allowed the evaluation of the contamination status, distribution and possible pollution sources. HCH, DDT, HCB, PCB and PAH were identified compounds in all the samples, whereas the concentrations of chlorinated pesticides (trans-eptachlorepoxide, dieldrin, endrin, metaoxychlor and mirex) were below the detection limits and were not of great concern. The pesticides did demonstrate markedly different distributions reflecting different agricultural and domestic usage in the region. The range of concentrations of HCH, HCB, DDT and PCBs in the sediments were 0.11-0.40, <0.05-0.98, 0.18-1.93 and 0.18-2.33 ng/g dry wt, respectively. Overall elevated levels of HCB, DDT and PCB were recorded at Babughat, a very closely located site of the metropolitan megacity Calcutta. Fluoranthene (flu) and pyrene (py), the dominant 4-ring PAHs, also showed an abrupt elevated concentrations at Babughat with values of 214 and 144 ng/g dry wt, respectively. Among the isomers and metabolites of HCH, DDT and PCB, alpha-HCH, pp'-DDT and PCB (101), PCB (118), PCB(153) and PCB (138) were found to be dominant. High ratios of metabolites of DDT to SigmaDDTs reveal recent use of DDT in this coastal environment. Fluoranthene, pyrene, 1,2-benzo(a)anthracene and chrysene formed the dominant congeners out of 19 identified PAHs. The sources of contamination are closely related to human activities such as domestic and industrial discharge, automobile exhausts, street runoff, slum sewage, agricultural chemicals and soil erosion due to deforestation as well as atmospheric transport. This study is compared to other coastal and estuarine environments in India and abroad. The baseline data can be used for regular ecological monitoring, considering the industrial and agricultural growth around this important estuarine ecosystem. PMID- 15788194 TI - Selecting peptide ligands of microcystin-LR from phage displayed random libraries. AB - In the present study, we investigated to find novel ligands for low molecular weight environmental toxin, microcystin-LR (MC-LR) by using phage display technology. Two random libraries, displaying linear 12-mer peptides and cyclic 7 mer peptides, were screened against the immobilized target respectively. After three rounds of panning, phage clones that recognized microcystin-LR specifically were obtained from both the linear and the constrained libraries, proved by enzyme-linked immumosorbent assays and immunoprecipitation assays. DNA sequencing indicated that peptides displayed on some of the selected clones shared consensus sequences. Compared with traditional methods, this approach provided a cheaper and more rapid alternative to screen specific ligands for microcystin-LR. Moreover, since it is rather difficult to take small molecules as targets of phage display libraries, the success of this experiment expanded the applications of phage display technology, and provided a new avenue to study environmental small molecular toxins. PMID- 15788195 TI - Effects of moderate pollution on toxic and trace metal levels in calves from a polluted area of northern Spain. AB - This study evaluated the contribution of anthropogenic pollution to toxic metal residues (Cd, Pb and As) in cattle in an industrialized area of Asturias (northern Spain), and investigated possible implications of toxic metal exposure for metabolism of essential metal elements (Cu, Zn, Fe, Mn). Samples of liver, kidney, muscle and blood from animals aged 9-12 months were obtained from an industrialized area (78 calves) and from a rural area (92 calves). Samples were acid-digested, and levels of metals determined by atomic absorption spectrophotometry. Cadmium and lead contents in the liver and kidney were moderately and significantly higher in calves from the industrialized area (Cd: liver 29.6, kidney 161; Pb: liver 38.1, kidney 38.3 microg/kg wet weight) than in calves from the rural area (Cd: liver 22.9, kidney 96.4; Pb: liver 20.7, kidney 15.9 microg/kg kg wet weight). Although these toxic metals were only moderately raised, our results suggest that they may nevertheless have interfered with trace element metabolism, as reported previously for severely polluted regions. Notably, copper levels in calves from the industrialized area were generally low, and nearly half of these animals showed tissue copper levels indicating risk of copper deficiency. PMID- 15788196 TI - Characteristics and health implications of fine and coarse particulates at roadside, urban background and rural sites in UK. AB - Recent studies have pointed to evidence that fine particles in the air could be significant contributors to respiratory and cardiovascular diseases and mortality. Epidemiologists looking at the health effects of particulate pollution need more information from various receptor locations to improve the understanding of this problem. Detailed information on temporal, spatial and size distributions of particulate pollution in urban areas is also important for air quality modellers as well as being an aid to decision and policy makers of local authorities. This paper presents a detailed analysis of temporal and seasonal variation of PM(10) and PM(2.5) levels at one urban roadside, one urban background and one rural monitoring location. Levels of PM(10), PM(2.5) and coarse fraction of particulates are compared. In addition, particulate levels are compared with NO(2) and CO concentrations. The study concludes that PM(10) and PM(2.5) are closely related at urban locations. Diurnal variation in PM(2.5)/PM(10) ratio shows the influence of vehicular emission and movement on size distribution. This ratio is higher in winter than in summer, indicating a build-up or longer residence time of finer particulates or washout due to wet weather in winter. In the second part of this study, a disease burden analysis is carried out based on the dose-response relationships recommended by the UK Committee on the Medical Effects of Air Pollution. The disease burden analysis indicates that if Marylebone Road (MR) levels of PM(10) were prevalent all over London, it will result in around 2.5% increase in death rates due to all causes. Whereas, if Bloomsbury (BB) levels were prevalent in London, which is more likely to occur as this is more representative of the urban background environment to which people in London are likely to be exposed, the corresponding increase would be around 1.7%. Considering this, in London, at Bloomsbury levels, 973 deaths and 1515 respiratory hospital admissions (RHA) are attributable to PM(10) while 2140 RHA are attributable to NO(2). After deducting the disease burden due to background levels at Rochester (RC), PM(10) emission caused by anthropogenic activities in London equates to 273 additional deaths and 410 additional RHA, while NO(2) account for additional 1205 incidences of RHA. PMID- 15788197 TI - World crop residues production and implications of its use as a biofuel. AB - Reducing and off-setting anthropogenic emissions of CO(2) and other greenhouse gases (GHGs) are important strategies of mitigating the greenhouse effect. Thus, the need for developing carbon (C) neutral and renewable sources of energy is more than ever before. Use of crop residue as a possible source of feedstock for bioenergy production must be critically and objectively assessed because of its positive impact on soil C sequestration, soil quality maintenance and ecosystem functions. The amount of crop residue produced in the US is estimated at 367x10(6) Mg/year for 9 cereal crops, 450x10(6) Mg/year for 14 cereals and legumes, and 488x10(6) Mg/year for 21 crops. The amount of crop residue produced in the world is estimated at 2802x10(6) Mg/year for cereal crops, 3107x10(6) Mg/year for 17 cereals and legumes, and 3758x10(6) Mg/year for 27 food crops. The fuel value of the total annual residue produced is estimated at 1.5x10(15) kcal, about 1 billion barrels (bbl) of diesel equivalent, or about 8 quads for the US; and 11.3x10(15) kcal, about 7.5 billion bbl of diesel or 60 quads for the world. However, even a partial removal (30-40%) of crop residue from land can exacerbate soil erosion hazard, deplete the SOC pool, accentuate emission of CO(2) and other GHGs from soil to the atmosphere, and exacerbate the risks of global climate change. Therefore, establishing bioenergy plantations of site-specific species with potential of producing 10-15 Mg biomass/year is an option that needs to be considered. This option will require 40-60 million hectares of land in the US and about 250 million hectares worldwide to establish bioenergy plantations. PMID- 15788198 TI - Uptake of polychlorodibenzo-p-dioxins, polychlorodibenzofurans and coplanar polychlorobiphenyls in chickens. AB - Twelve chickens raised according to commercial standards were fed a diet containing about 30 ng total toxic equivalent (TEQ)/kg for 10 weeks. Persistent pollutants were introduced into the poultry feed via recycled oil to mimic contamination conditions closely resembling those occurring during the Belgian crisis five years ago. Absorption of congeners with the same chlorination degree did not seem to depend on the substitution, demonstrating that unlike for cows, no preferential absorption for 2,3,7,8-substituted compounds could be remarked for chickens. As already observed, absorption decreased with increasing number of chlorines and was not linearly dependent on the octanol/water partition coefficient. On the other hand, no real differences were observed in the absorption of coplanar polychlorinated biphenyls (c-PCBs) with regard to degree of chlorination. When monitored during the course of experiment, concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and c-PCBs in excreta reached an apparent steady state after 5 weeks. Only 2,3,7,8-substituted dioxins or furans were found in tissues and eggs. All organs showed the same congener profile and similar lipid-normalized concentration, except for the liver. Bioconcentration factors were evaluated, highlighting that the liver preferentially retained highly chlorinated congeners. No depletion of dioxin and PCB concentration was observed after 8 and 14 weeks of control diet, but high inter-individual variation occurs. PMID- 15788199 TI - Geochemistry and bioavailability of metals in sediments from northern San Francisco Bay. AB - In this study, metals (Be, Cr, Mn, Fe, Ni, Cu, Zn, Ag, Cd, Pb and Hg) in the fine grained fraction (<63 microm) from 12 sites at different locations in northern San Francisco Bay over a year period from March 2000 to March 2001 were analyzed after acid extraction. The results showed that metal concentrations in the sediments varied from site to site, whereas some of them were found elevated with respect to the sediment of Tomales Bay, CA, which has little contamination history, indicating an enrichment of the metals in the sediment samples analyzed. Sediment toxicity and bioaccumulation evaluation by a clam species, Macoma nasuta, exposed to the sediment samples collected from the six sampling sites was carried out. The results showed that the sediment samples tested significantly reduced clam survival. Toxicity of the sediments to the clam was, in part, related to elevated metal concentrations in the sediments. In order to examine geochemistry of the metals and to understand potential correlations between metal concentrations and geochemical matrix elements of the sediments, bioavailability and toxicity of the metals, detailed analysis of metal concentrations associated with total organic carbon and the Fe-oxy-hydroxides in the sediment samples was performed. The analysis showed that sediment geochemistry appeared to influence metal bioavailability and may have important impacts on the toxicity of these metals to the clam. PMID- 15788200 TI - Effects of dissolved organic matter on toxicity and bioavailability of copper for lettuce sprouts. AB - It is well known that dissolved organic matter in soil solution may affect the toxicity or bioavailability of heavy metals to plants, but existing information on various organic substances is insufficient for treating problems with heavy metal-contaminated soils. To clarify how dissolved organic matter alters the toxicity and bioavailability of metals, we germinated lettuce seeds exposed to solutions containing Cu and several kinds of dissolved organic matters. Low molecular weight organic acids (citric, malic, and oxalic acids) increased the toxicity and bioavailability of Cu, but low concentrations of the synthetic chelators ethylenediamine tetra-acetic acid (EDTA) and diethylenetriamine penta acetic acid (DTPA) decreased the toxicity and bioavailability of Cu. In contrast, humic acid appeared to be the most effective organic substance for detoxifying Cu, even though it did not significantly decrease the bioavailability of Cu. Consequently, the bioavailability and toxic effects of Cu in soil depend on the nature of coexisting organic substances in the soil solution. PMID- 15788201 TI - Cadmium phytoextraction using short-rotation coppice Salix: the evidence trail. AB - A substantial body of evidence has now accumulated that raises expectations that clean-up of Cd-contaminated land can be achieved through cultivation and harvest of selected clones of short-rotation coppice willow within a realistic crop lifecycle. Cd uptake rates into Salix are high compared to other trace elements and to other plant species. Effective phytoextraction would require (i) careful targeting of hotspots, (ii) repeated harvest prior to leaf fall and (iii) final removal of the root bole. PMID- 15788202 TI - Daphnia emergence: a sensitive indicator of fire-retardant stress in temporary wetlands. AB - Fire-retardant formulations are increasingly used by fire managers to control wildland fires. Their extensive use requires an assessment of their impacts in those ecosystems that could be affected by them. Recent studies indicate the potential for environmental impacts when accidentally delivered to surface waters. Yet the response of temporary wetlands, such as vernal pools, typical in Mediterranean areas, is unknown. This study reports on the emergence response of a wetland population of Daphnia curvirostris Eylmann (Cladocera, Crustacea) from sediments that were treated with a commercial fire retardant (Fire-Trol 934). Three application levels were used: 1, 3 and 5 L m(-2). The low and medium levels are in the range of manufacturer's recommendations of use in the field based on the fuel characteristics. The high level simulates local elevated concentrations that may result from inhomogeneous retardant delivery. Results indicate that emergence success decreases with increasing application rate, leading to a complete failure with application levels of 5 L m(-2). This suggests that benthic pelagic interactions in temporary wetlands-often the primary way by which recolonization of isolated wetlands occurs-can be significantly impacted by fire retardants once they fill with fall/winter rains, thereby serving as a sensitive indicator of fire-retardant contamination. Results highlight a research need for establishing of criteria for effective, but environmentally safe use of fire retardants in the environment. PMID- 15788203 TI - Hepatitis C virus and non-Hodgkin's lymphoma 10 years later. AB - Hepatitis C virus is associated with chronic liver disease as well as with lymphoproliferative disorders such as mixed cryoglobulinemia and, likely, non Hodgkin's lymphomas. The association between hepatitis C virus infection and B cell lymphoma is controversial since it shows a strong regional variation. In fact, the prevalence of hepatitis C virus infection in non-Hodgkin's lymphoma shows a prevalence ranging between 7.4 and 37.0%. However, the intimate pathogenetic mechanism involved in hepatitis C virus-associated lymphomas remains considerably unknown. Hepatitis C virus may exert its oncogenic potential via an indirect mechanism or utilise other pathways directly. It is reasonable to assume that several different pathogenetic mechanisms operate in the wide spectrum of hepatitis C virus-related lymphoproliferative disorders, which include the intermediate to high-grade lymphoma, and the more common indolent, low-grade lymphoma, preceded by long standing symptomatic mixed cryoglobulinemia Type II. In this review, the etiopathogenetic role of hepatitis C virus in non-Hodgkin's lymphoma is discussed on the basis of molecular, clinical and epidemiological considerations. The management of hepatitis C virus-associated non-Hodgkin's lymphoma is similar to that of conventional lymphoma, although viral reactivation or the underlying chronic liver disease can complicate chemotherapy. Whether to treat low-grade hepatitis C virus-related lymphomas with anti-viral therapy is still debatable, but encouraging data emerge from some recent studies. PMID- 15788204 TI - Einstein, Darwin and conventional wisdom about Crohn's disease. PMID- 15788205 TI - Treatment with inhibitors of angiogenesis in advanced hepatocellular carcinoma: a new tool in our hands or simply a hope? PMID- 15788206 TI - Increased phospholipase activity in Helicobacter pylori strains isolated from patients with gastric carcinoma. AB - PURPOSE: Phospholipase activity, one of Helicobacter pylori pathogenicity factors, has not been investigated enough, so far, although it may induce a remarkable damage to the gastric mucosa. In the present work, we have compared the whole phospholipase activity of H. pylori strains isolated from patients with gastric carcinoma with that of strains isolated from dyspeptic patients without gastric carcinoma. METHODS: We measured the phospholipase activity of one distinct H. pylori colony isolated from each of 10 patients with gastric carcinoma and 10 controls, dyspeptic patients without endoscopic and histological signs of gastric carcinoma. We also determined the phospholipase activity of 20 additional strains isolated from different areas of neoplastic and non-neoplastic tissue of two patients with gastric carcinoma, the cagA and vacA positive G27 and 328 wild strains and their respective vacA and cagA negative isogenic mutants. The whole phospholipase activity of strains was determined by measuring the release of (14)C-labeled palmitic acid from the radioactive l-3 phosphatidylcholine, 1,2-di[1-(14)C]palmiloyl substrate; results were expressed in pmol of palmitic acid per mg of protein. RESULTS: H. pylori strains isolated from patients with gastric carcinoma had levels of phospholipase activity significantly higher than those of strains isolated from controls (99.37 [S.D. 40.45] versus 34.46 [S.D. 16.46], P<0.001). In patients with gastric carcinoma, the mean phospholipase activity of strains isolated from neoplastic tissue was similar to that of strains isolated from non-neoplastic tissues (123.02 [S.D. 44.36] and 115.77 [S.D. 81.48], respectively. Interruption of cagA gene caused a ca. 20% reduction of phospholipase activity (36.38 versus 45.22 of the wild strain); that of vacA caused no reduction of phospholipase activity (26.53 and 25.37 of the wild strain). CONCLUSIONS: The infection by H. pylori strains that produce high levels of phospholipase may increase the risk of developing gastric carcinoma. We hypothesise that indirect products of phospholipase activity, such as prostaglandins, leukotrienes and lysophospholipids, may mediate carcinogenesis. PMID- 15788207 TI - Outcomes after adult isolated small bowel transplantation: experience from a single European centre. AB - BACKGROUND: Adult isolated small bowel transplantation is considered the standard treatment for patients with life-threatening parenteral nutrition-related complications. Here, we report a 3-year experience in a single European centre between December 2000 and December 2003. AIMS: To evaluate and discuss pre transplant and post-transplant factors that influenced survival rates in our series. PATIENTS: Fourteen patients, with a mean parenteral nutrition course of 27 months, were transplanted. In eight cases they had not experienced any major complication from parenteral nutrition. METHODS: We described pre-transplant evaluation and inclusion criteria, surgical technique and clinical management after transplant. Immunosuppressive therapy was based on induction drugs and Tacrolimus. We reported survival rates, major complications and rejection events. RESULTS: One-year actuarial survival rate was of 92.3% with a mean 21-month follow-up (range 3-36 months). We had no intraoperative deaths. One patient (7.2%) died of sepsis following cytomegalovirus enteritis. One patient underwent graftectomy (7.2%) for intractable severe acute rejection. One-year actuarial graft survival rate of 85.1%. One patient (7.2%) affected by post-transplant lymphoproliferative disease is alive and disease-free after 8 months. CONCLUSION: We believe candidate selection, induction therapy, donor selection and short ischemia time play an important role in survival after small bowel transplantation. PMID- 15788208 TI - Evolution of clinical behaviour in Crohn's disease: predictive factors of penetrating complications. AB - BACKGROUND AND AIMS: Crohn's disease is a heterogeneous entity. The Vienna Classification defines three different clinical patterns: 'non-stricturing, non penetrating', 'stricturing' and 'penetrating'. Aim of this study was to assess the change in clinical behaviour over time and to evaluate whether an evolution towards penetrating complications can be predicted. METHODS: A total of 139 patients with non-penetrating behaviour at the time of diagnosis were included. The mean follow-up was 4.84 years (range 1-23.2 years). The clinical behaviour, according to the Vienna Criteria, was assessed at the diagnosis and at the end of follow up. Statistical analysis was performed by means of the Kaplan-Meier method and standard logistic regression analysis. RESULTS: The cumulative probability of a change in clinical behaviour was 22, 38 and 63% at 3, 6 and 12 years, respectively, and the cumulative probability of developing penetrating complications was 22, 33 and 55% at 3, 6 and 12 years, respectively. Young age at diagnosis (<40 years) and a stricturing behaviour are independent risk factors of developing major penetrating complications (internal fistula, mass or abscess): OR=6.0, 95% CI 1.1-30.5; OR=4.0, 95% CI 1.5-10.9, respectively, but not perianal disease. CONCLUSIONS: The behavioural classification of Crohn's disease is a dynamic model in which each status should be considered as not fixed but evolutive. Perianal disease should be considered a distinct pattern of penetrating behaviour. PMID- 15788210 TI - Screening for hepatocellular carcinoma. AB - BACKGROUND AND AIM: The incidence of hepatocellular carcinoma is high among cirrhotic patients, ranging from 2 to 3% in western cohorts and 6-11% in eastern cohorts. Although only one randomised trial has been performed, clinical practice generally uses periodic screening to detect hepatocellular carcinoma in cirrhotic patients. We reviewed the scientific literature on hepatocellular carcinoma screening. MATERIALS AND METHODS: Evaluation of studies identified through MEDLINE and EMBASE (1990-May 2003). RESULTS: The available screening tests to detect hepatocellular carcinoma are alpha-fetoprotein (cut-off: 20 ng/ml) and ultrasound, which are generally combined. The reported sensitivity and specificity are 50-85% and 70-90%, respectively. An estimated doubling time of about 6 months has led to the use of an interval of 6 months between screenings. Based on the risk of hepatocellular carcinoma, cirrhotic patients are considered as the target population. Screening seems to detect smaller and more frequently unifocal hepatocellular carcinoma; the residual liver function is important for determining the eligibility for effective treatment (resection); hence the prevention is more effective for patients with well-compensated cirrhosis. The survival estimated by non-randomised studies is slightly longer for patients with screening-detected hepatocellular carcinoma, compared to those with clinically detected hepatocellular carcinoma, although few studies have accounted for 'lead time bias'. CONCLUSIONS: Although screening for the early detection of hepatocellular carcinoma has become quite common in clinical practice, its effectiveness remains controversial. Observational studies that have taken into account lead time bias suggest that survival is greater for patients with screening-detected hepatocellular carcinoma, yet the eligibility for effective treatments is low. Considering that only one randomised controlled trial has been conducted, it is crucial to standardise the screening schedule and to evaluate prevention programmes. PMID- 15788209 TI - Effects of the combined treatment with thalidomide, megestrol and interleukine-2 in cirrhotic patients with advanced hepatocellular carcinoma. A pilot study. AB - BACKGROUND: Thalidomide, an anti-angiogenic agent, does not have a good therapeutic effect for advanced hepatocellular carcinoma when used alone. Megestrol and interleukin-2 have been proposed as a palliative treatment for hepatocellular carcinoma. AIMS.: We assessed the effectiveness/safety of a combined therapy with thalidomide+megestrol+interleukin-2 in cirrhotic patients with advanced hepatocellular carcinoma. PATIENTS AND METHODS: Nine cirrhotic patients with advanced hepatocellular carcinoma received oral megestrol (160 mg/day) and thalidomide (from 50 mg/day to the maximal tolerated dose). Four patients also received subcutaneous interleukin-2 (1 million U/day for 21 days/month). RESULTS: The maximal tolerated dose of thalidomide was 150 mg/day. All patients complained of sedation and other neurological or digestive adverse effects. In all but one patient the adverse effects disappeared after thalidomide withdrawal or dose reduction. Interleukin-2 administration caused a flu-like syndrome and a reaction at the injection site. During treatment, alpha fetoprotein increased in six patients, remained stable in two and decreased in one. Eight patients showed tumour progression and one had a stable disease. Eight patients died. The median survival was 9.9 (range 2.6-18.6) months. CONCLUSION: In cirrhotic patients, the combined treatment with thalidomide+megestrol (+/ interleukin-2) does not control hepatocellular carcinoma growth, possibly due to the low tolerance to thalidomide and interleukin-2 preventing the use of appropriate dosages. PMID- 15788211 TI - Expression of connective tissue growth factor is a prognostic marker for patients with intrahepatic cholangiocarcinoma. AB - BACKGROUND AND AIMS: Connective tissue growth factor is a member of the 'CCN' protein family. Consistent with its profibrotic properties, it is over-expressed in several human epithelial malignancies. PATIENTS AND METHODS: We have retrospectively evaluated by immunohistochemistry the presence of connective tissue growth factor in archival tissues from 55 resected intrahepatic cholangiocarcinomas and compared its expression to the main pathological parameters, disease free and overall survival. RESULTS: Tumours were scored as high and low/absent expressers (> or =50%, 0-50% cells, respectively). Thirty three of 55 cholangiocarcinomas (60%) were high and 22 (40%) low expressers. No significant correlation was found between connective tissue growth factor and tumour grade, tumour location, vascular and perineural invasion. Eighteen of 22 (82%) low/absent expressers and 12/33 (36%) high expressers had recurrence of disease (P=0.001). Low/absent expressers showed a poor disease free and overall survival compared with the higher expressers (P<0.001). Vascular invasion was related to tumour recurrence (P=0.025) and to decreased disease free survival (P<0.05). During proportional hazard regression analysis, only connective tissue growth factor was found to influence disease free survival (P=0.01). CONCLUSIONS: Expression of connective tissue growth factor is an independent prognostic indicator of both tumour recurrence and overall survival for intrahepatic cholangiocarcinoma patients regardless of tumour location, tumour grade, vascular and perineural invasion. PMID- 15788212 TI - A case of gastrointestinal stromal tumour of the ampulla of Vater. AB - Gastrointestinal stromal tumour rarely develops in the duodenal ampulla region. We report here a case of gastrointestinal stromal tumour of the ampulla of Vater found in a 44-year-old Japanese man presenting with biliary obstruction. He died of hepatic failure with diffuse liver metastasis. The postmortem examination showed a large Borrman type III-like tumour in the duodenal ampullary region with direct invasion of the pancreas and extrahepatic bile duct as well as metastases to the liver and regional lymph nodes. The duct orifice was located at the centre of the tumour. Microscopically, the tumour consisted of anaplastic spindle cells with high mitotic activity (90 mitoses per 50 high-power fields). Immunohistochemically, the spindle cells were positive for KIT and CD34. The final diagnosis was high-grade malignant gastrointestinal stromal tumour of the ampulla of Vater. Considering the recent advances in the diagnosis and treatment of gastrointestinal stromal tumour, this neoplasm should be included in the differential diagnosis of the tumours appearing in the duodenal ampulla region. PMID- 15788213 TI - Presacral carcinoid tumour. Review of the literature and report of a clinically malignant case. AB - Carcinoid tumours arising in the presacral region are extremely rare and they are usually benign. We report the case of a 37-year-old black man with a clinically malignant carcinoid tumour (well differentiated endocrine carcinoma) occurring in a sacrococcygeal teratoma and already metastasised to pelvic nodes, liver and bone at the time of the initial diagnosis. Such an aggressive behaviour of the presacral carcinoid tumours has never been described. The derivation of these tumours from hindgut rests with reference to embryological development of the tailgut cysts is discussed. PMID- 15788214 TI - Thiopurines in inflammatory bowel disease: pharmacogenetics, therapeutic drug monitoring and clinical recommendations. AB - There is a growing interest in the use of thiopurines (azathioprine, 6 mercaptopurine and 6-thioguanine) for the management of inflammatory bowel disease. The genetically controlled thiopurine (S)-methyltransferase enzyme is involved in the metabolism of these agents and is hypothesised to determine the clinical response to thiopurines. Diminished activity of this enzyme decreases the methylation of thiopurines, theoretically resulting in potential overdosing, while high thiopurine (S)-methyltransferase status leads to overproduction of toxic metabolites and ineffectiveness of azathioprine and 6-mercaptopurine. In practice, controversies exist regarding the utility of standard thiopurine (S) methyltransferase pheno- and genotyping. Current pharmacogenetic insights suggest that another enzyme system may participate in the efficacy and toxicity of thiopurines; inosine triphosphate pyrophosphatase. Other topics discussed in this review are the utilisation of therapeutic drug monitoring and the experimental use of 6-thioguanine in the treatment of inflammatory bowel disease. 6 Thioguanine has a less genetically controlled metabolism and skips genetically determined metabolic steps. On theoretical basis, 6-thioguanine might therefore have a more predictable profile than azathioprine and 6-mercaptopurine. However, the use of 6-thioguanine has been associated with an increased risk of nodular regenerative hyperplasia of the liver and veno-occlusive disease. Further research is warranted before 6-thioguanine can be considered as a treatment option for inflammatory bowel disease. PMID- 15788215 TI - Coeliac disease and depression: two related entities? PMID- 15788216 TI - Hit rates of adequate performance based on the test of memory malingering (TOMM) Trial 1. AB - The Test of Memory Malingering (TOMM) is a method of identifying patients who may be exerting poor effort during neuropsychological testing. This study seeks to examine whether TOMM Trial 1 scores are useful in identifying patients whose overall TOMM performance does not indicate obvious poor effort. Hit rates of adequate performance on Trial 2 and the Retention Trial were calculated based on a fixed criterion of 45 or greater on Trial 1. Archival data were collected from a sample of 77 mild traumatic brain injured litigants and compared to each clinical sample described in the TOMM test manual (Tombaugh, 1996). Results show a hit rate of 100% for each group. Findings reveal that adequate Trial 1 scores predict adequate Trial 2 and Retention Trial scores, suggesting that patients scoring 45 or greater on Trial 1 are not likely to be suspected of poor effort based on overall TOMM performance. PMID- 15788217 TI - Discrepancies between self-reported years of education and estimated reading level: potential implications for neuropsychologists. AB - Current standard neuropsychology practice is to examine normative sample performance for systematic influences of demographic variables and then to correct for these influences. The most commonly examined demographic variables are age, gender, and years of education, and current normative databases frequently take these into consideration. However, there is a literature to suggest that self-reported years of educational attainment may not be an accurate reflection of some patients' level of performance and may actually overpredict grade estimates based on reading level. Many of these studies have focused on older samples of individuals who were free of neurological or psychiatric symptoms. In this study, a younger sample (average age = 44.5) of African American (N = 62) and Caucasian (N = 133) patients referred to an outpatient psychiatry unit was examined. Results suggest that the prior findings of a significant discrepancy between self-reported years of education and Wide Range Achievement Test-3rd Edition Reading Recognition performance hold for a younger sample with a broad range of clinical diagnoses. How these findings might influence clinical practice are discussed. PMID- 15788218 TI - Treating dementia patients with vascular lesions with donepezil: a preliminary analysis. AB - Sixteen patients with Alzheimer's disease (AD) and 15 patients with vascular dementia (VaD) associated with subcortical white matter lesions or subcortical cardiovascular accidents (CVAs) were treated with donepezil for 16 weeks. Within group analyses for the AD group revealed significant improvement on some tests of working memory, and marginal improvement on the Mini-Mental State Examination and on tests of immediate free recall from a serial list-learning task. Identical analyses for the VaD group revealed substantial gains on tests of working memory and delayed recognition memory. These data suggest that medication such as donepezil may act to improve the working memory deficits known to be associated with dementia patients with subcortical vascular lesions. The clinical implications of these findings are discussed. PMID- 15788219 TI - Psychometric properties of the benton visual form discrimination test. AB - Coefficient alpha and an item analysis were calculated for the 16-item Benton Visual Form Discrimination Test (VFDT) using a heterogeneous sample (N = 293) of mostly elderly medical patients who were suspected of having cognitive impairment. The total score reliability was .74. An item analysis found that 15 of the items were within established criteria for item difficulty, however, 5 items were found to be poor discriminators. Through the use of confidence intervals around observed scores, it was shown that the current classification criterion for the VFDT demands a higher reliability coefficient than what was found. Also, evidence for the test's insufficient level of difficulty is presented. It is difficult to recommend this test for clinical use. PMID- 15788220 TI - Performance of community-dwelling adults ages 55 to 75 on the University of Pennsylvania smell identification test: an item analysis. AB - Tests of odor identification, such as the University of Pennsylvania Smell Identification Test (UPSIT; Doty, 1995), are used in both research and clinical settings to assess the individual's sense of smell. Although previous studies have evaluated the psychometric properties of the UPSIT, little research has been conducted explicitly with unimpaired adults. This study evaluated the results of the UPSIT for 197 community-dwelling adults between the ages of 55 and 75. An item analysis identified six items that had poor hit rates in this sample. Implications of these results are discussed. PMID- 15788221 TI - Symptom checklist-90 revised scores in persons with traumatic brain injury: affective reactions or neurobehavioral outcomes of the injury? AB - The goal of this study was to examine the concurrent validity of the Symptom Checklist-90 Revised (SCL-90-R) as a measure of emotional distress among persons with traumatic brain injuries (TBI). Following previous studies, the scale was divided into a "Brain Injury Subscale" (BIS), composed of items that are confounded with the neurobehavioral outcomes of TBI, and a "Non Brain Injury Subscale" (NBIS), composed of items unrelated to the neurobehavioral outcomes. The scores of 94 persons with TBI were analyzed on the two subscales. Although more frequently endorsed, the BIS items were equally related to the cognitive and behavioral outcomes of the injury and to the respondents' affective dispositions. The same pattern of correlations was evident with the NBIS items. In addition, both scales were predicted by measures of emotional reactions to the injury. These results were interpreted as supporting the validity of the SCL-90-R as a measure of emotional distress among persons with brain injuries. PMID- 15788222 TI - Judgment of emotional nonlinguistic vocalizations: age-related differences. AB - Humans make extensive use of vocal information to attribute emotional states to other individuals. To date, most studies exploring perception of vocal emotions have done so in the context of speech prosody, although nonlinguistic emotional vocalizations represent an important, perhaps more universal, means to express emotions. Here, we explored the perception of emotional nonlinguistic vocalizations in healthy individuals, with an emphasis on potential age- and sex related differences. Sixty participants rated 563 positive (e.g., laughs, sexual vocalizations), negative (e.g., cries, screams of fear), and neutral vocalizations (e.g., coughs), according to the valence, intensity, and authenticity of the emotion expressed. Ratings were consistent among individuals, suggesting that valence is an adequate measure of emotional categorization. An important effect of age emerged: (a) age by vocalization category interactions were observed for both valence and intensity ratings, and (b) younger participants rated stimuli as more emotional than older individuals (i.e., higher valence for positive, lower for negative, and more intense for both positive and negative). We also found a sex effect in the authenticity ratings: older women rated the vocalizations as less authentic than younger women whereas authenticity judgments did not differ between the two age groups in men. Taken together, these findings suggest that, as previously observed for facial expressions and prosody, the judgments of emotional vocalizations may vary with age. PMID- 15788223 TI - Alternating hemiplegia of childhood: a study of neuropsychological functioning. AB - Alternating hemiplegia of childhood (AHC) is a rare chronic neurological disorder characterized by alternating transient attacks of hemiplegia. Evidence from recent studies suggests the disorder is characterized by tonic fits, dystonic posturing, and ocular motor abnormalities. Although widely observed, deficits in cognitive functioning are among one of the least described features of AHC. Most studies describe global deficits, however, no study has provided a comprehensive neuropsychological assessment of a child with AHC. This study presents neuropsychological findings from a comprehensive assessment of a single child with AHC. Results showed significant deficits in intellectual, academic, memory, attention, and executive functioning. Additional deficits in language, psychomotor abilities, and psychosocial functioning were noted. Preliminary longitudinal data are also offered to examine the progressive nature of this disorder. Cognitive strengths and weaknesses are discussed at length as well as recommendations for the classroom and future research. PMID- 15788225 TI - The influence of biosurfactants released by S. mitis BMS on the adhesion of pioneer strains and cariogenic bacteria. AB - The influence of Streptococcus mitis BMS biosurfactants on the adhesion of eight pioneer and four cariogenic oral bacterial strains was, for a first screening, examined in a microtiter plate assay. The adhesion to pellicle-coated wells of three cariogenic strains was inhibited >70% by the biosurfactants, while only one pioneer strain showed >70% reduction. The reduction for the other strains did not exceed 50%. Subsequently, adhesion of Streptococcus mutans ATCC 25175 and Streptococcus sobrinus HG 1025, both cariogenic strains, and Actinomyces naeslundii T14V-J1 and Streptococcus oralis J22, two pioneer strains, to biosurfactants-coated enamel with and without a salivary pellicle was studied in a parallel plate flow chamber. A biosurfactants coating to enamel with or without a pellicle caused a reduction in the number of adhering cariogenic organisms, although no such reduction was observed for the pioneer strains. Consequently, it is concluded that S. mitis BMS biosurfactants may play a protective role against adhesion of cariogenic bacteria. PMID- 15788226 TI - Human palatal saliva: adsorption behaviour and the role of low-molecular weight proteins. AB - In situ ellipsometry was employed to study adsorption from human palatal saliva (HPalS) in terms of dependence on surface wettability and saliva concentration (60 years) (p=0.02). Patients with hemoglobin concentration>13.4 g/dl before therapy (median hemoglobin concentration) had a significantly better overall survival than patients with lower hemoglobin concentration (p=0.044). Patients who received combined radiochemotherapy (with or without operation) had a survival advantage compared with radiotherapy alone. Overall survival after neoadjuvant treatment followed by operation was significantly better than in the two other groups, median survival times were 20 vs. 9 (RCHT) vs. 8 months (RT) (p=0.003). The data presented show for the first time that hemoglobin concentration in addition to gender and age was a prognostic factor for patients with esophageal cancer. A low hemoglobin value was a negative predictor. PMID- 15788298 TI - Concurrent presentation of stage I teratoma and stage I renal cell cancer. PMID- 15788299 TI - Delayed radiation myelopathy after concurrent chemoradiation for hypopharyngeal esophageal carcinoma. PMID- 15788303 TI - Maternity and bone mineral density. AB - During pregnancy and lactation, changes occur in a variety of factors which have great potential to influence bone mineral density (BMD). Smoking habits, the level of alcohol consumption, the level of physical activity, body weight, soft tissue composition and hormone levels are all factors that change during the course of these conditions. Some of these factors are capable of increasing BMD, and some can reduce it. Due to these various changes, it is virtually impossible to predict the development in BMD that will occur during a pregnancy and lactation. However, longitudinal studies have suggested that both pregnancy and lactation are associated with a BMD loss of up to 5%, albeit that the BMD recovers after weaning. Cross-sectional studies have indicated that women with many children and a long total period of lactation have similar or higher BMD and similar or lower fracture risk than their peers who have not given birth. As the studies showing this trend have been observational and cross sectional case control studies, the conclusions can only be regarded as being suggestive, and no causality can be proven. PMID- 15788304 TI - Hand transplantation. A future clinical option? AB - Transplantation of a vascularized limb or its components is defined as composite tissue allotransplantation, and is one of the newest areas in surgery. To date, 24 hands have been transplanted onto 18 recipients. The initial results have been promising, and hand transplantation may become an important procedure for functional restoration of upper limbs. However, the ethical aspects of using chronic immunosuppression for a condition which is not life threatening have been the subject of debate. In this article, we review the field of composite tissue allotransplantation. PMID- 15788305 TI - Total hip arthroplasty for primary osteoarthrosis in younger patients in the Finnish arthroplasty register. 4,661 primary replacements followed for 0-22 years. AB - BACKGROUND: Many studies have found a higher risk of revision after hip arthroplasty in younger patients. We evaluated the population-based survival of total hip arthroplasty (THA) in patients under 55 years of age and the factors affecting survival. METHODS: The Finnish Arthroplasty Register was established in 1980, and 74,492 primary THAs were entered into the register between 1980 and 2001. 4,661 of these were evaluated, all of which had been performed for primary osteoarthrosis on patients under 55 years of age. RESULTS: Proximally circumferentially porous-coated uncemented stems implanted between 1991 and 2001 had a 10-year survival rate of 99 (95% CI 98.5-99.6)% with aseptic loosening as endpoint. The risk of stem revision due to aseptic loosening was higher in cemented stems than in proximally porous-coated (RR 5.5, p < 0.001) or HA-coated (RR 6.6, p = 0.01) uncemented stems implanted during the same period. According to Cox regression analysis of cups implanted 1991-2001, the risk of revision for all-polyethylene cemented cups was 3.0 times as high as that for press-fit porous coated uncemented cups with aseptic loosening as endpoint (p = 0.01). However, when the endpoint was defined as any revision (including exchange of liner), there was no longer any difference between these two concepts, the 10-year survival rates being 94 (92.1-95.5)% for press-fit porous-coated uncemented cups and 93 (88.5-97.6)% for all-polyethylene cemented cups (p = 0.9). INTERPRETATION: Modern uncemented stems seem to have better resistance to aseptic loosening than cemented stems in younger patients. Thus, for younger patients, uncemented proximally circumferentially porous- and HA-coated stems are the implants of choice. Press-fit porous- and HA-coated uncemented cups may have better endurance against aseptic loosening than cemented cups in younger patients. However, when all revisions (including exchange of liner) are taken into account, the survival of modern uncemented cups is no better than that of all-poly cemented cups. PMID- 15788306 TI - Uncemented Harris-Galante total hip arthroplasty in patients with osteonecrosis of the femoral head. A 10-16-year follow-up study. AB - BACKGROUND: There have only been a few reports regarding the long-term results of uncemented THAs in patients with osteonecrosis. PATIENTS AND METHODS: We evaluated the long-term results of 65 consecutive uncemented total hip arthroplasties (Harris-Galante type I prostheses) in 54 men (59 hips) and 5 women (6 hips) with osteonecrosis of the femoral head. The mean age was 53 (33-64) years. The mean duration of follow-up was 12.5 (10-16) years. RESULTS: 9 femoral stems and 3 acetabular metal shells were revised. 2 polyethylene liners were changed due to excessive wear. A girdlestone procedure was done in 2 patients (2 hips) due to delayed deep infection. The mean polyethylene wear was 0.14 mm per year. 3 non-revised hips had pelvic osteolysis and 18 had femoral osteolysis. The 15-year survival rates, using failure defined as the removal of any component for any reason, were 85 (95% CI; 79-91)% for the acetabular and 80 (74-86)% for the femoral component and 70 (63-77)% for any of the components. INTERPRETATION: The first generation of the HGP design was frequently associated with pain, unstable fixation, and osteolysis. Excessive wear was frequent. The cup showed better durability than the stem. PMID- 15788307 TI - Higher failure rate and stem migration of an uncemented femoral component in patients with femoral head osteonecrosis than in patients with osteoarthrosis. AB - BACKGROUND: Several authors have found poorer outcome after hip replacement for osteonecrosis than after hip replacement for arthrosis. In a retrospective study we evaluated the performance of an uncemented femoral component in patients with osteonecrosis and arthrosis of the hip. PATIENTS AND METHODS: 31 patients operated for osteonecrosis, and 49 patients operated for osteoarthrosis were included. The median follow-up time was 6.1 (2-11) years for the patients with osteonecrosis, and 5.9 (4-8) for the arthrosis patients. RESULTS: Migration analysis performed by the Einzel-Bild-Roentgen Analysis (EBRA) technique revealed a median stem migration of 1.5 (-8.8-0) mm in the patients with osteonecrosis, but only 0.6 (-2.8-0.7) mm in the patients with arthrosis (p < 0.001). Survivorship analysis with stem revision as endpoint for failure was 74% (95% CI: 55-94) in the osteonecrosis, and 98% (95% CI: 94-100) in the arthrosis group (p = 0.01). INTERPRETATION: We suggest that the higher failure rate and stem migration of uncemented total hip replacement in the patients with osteonecrosis is a consequence of the disease. On the basis of these findings, we recommend close monitoring of the patients with osteonecrosis, which should include migration measurements. PMID- 15788308 TI - Young age and wedge stem design are risk factors for periprosthetic fracture after arthroplasty due to hip fracture. A case-control study. AB - BACKGROUND: It has been suggested that a hip fracture as a primary diagnosis for arthroplasty is a risk factor for periprosthetic fracture. PATIENTS AND METHODS: We compared 16 patients with late periprosthetic fracture of the femur to 48 controls. The primary diagnosis for all patients was hip fracture. The date of operation for hip fracture was used to select the control group of patients, all of whom were operated at the same time and in the same hospital district. No other selection criteria were used. Complications at the time of index operation, type of prosthesis, age, sex, BMI, patient's diseases, medication, and surgeon's experience were compared using conditional logistic regression. RESULTS: We found that low age at the time of the hip fracture operation increased the risk of periprosthetic fracture (OR = 4.9, CI = 1.2-20). The polished wedge type of prosthesis was associated with a higher risk of periprosthetic fracture than were other designs (OR = 11, CI = 1.2-97). INTERPRETATION: These findings of risk factors for periprosthetic fracture are new and should be confirmed by further studies using larger numbers of patients. PMID- 15788309 TI - Plasma levels of platelet-derived growth factor BB and transforming growth factor in patients with failed hip prostheses. AB - BACKGROUND: The role of growth factors in prosthesis loosening is unclear. We evaluated the levels of platelet-derived growth factor BB (PDGF-BB), transforming growth factors beta1 (TGF-beta1) and beta2 (TGF-beta2), both before and after activation, in patients with aseptic loosening of their hip prosthesis. PATIENTS AND METHODS: 26 patients with loosened hip implants were compared with 21 patients who had stable hip prostheses, and 28 patients undergoing primary hip replacement. The plasma levels of the growth factors were analyzed by enzyme immunoassay. TGF-beta1 and TGF-beta2 were determined both before and after activation. RESULTS: Patients with aseptic loosening had significantly lower PDGF BB levels than patients undergoing primary hip replacement, and significantly lower TGF-beta2 levels than patients with a stable implant. Patients with stable prostheses had significantly higher TGF-beta1 and TGF-beta2 levels than patients undergoing primary hip replacement. INTERPRETATION: It is possible that the prosthetic implant itself causes a local increase in PDGF-BB, TGF-beta1 and TGF beta2, released by osteoblasts and other cells in the microenvironment. The plasma PDGF-BB measured does not correspond to local release, which is probably due to local consumption or degradation. The consumption of PDGF-BB is low in stable implants, and TGF-beta1 and TGF-beta2 levels increase during bone formation. In loosening, PDGF-BB consumption is higher and causes a significant reduction in plasma levels as compared to presurgery. The formation of poor quality bone may be related to the scarce increase in TGF-beta1 and TGF-beta2. In conclusion, compared with patients with a stable implant, a reduction in bone forming growth factors appears to occur in individuals with aseptic loosening. PMID- 15788310 TI - Periacetabular osteotomy. Good pain relief in symptomatic hip dysplasia, 32 patients followed for 4 years. AB - BACKGROUND: When surgical treatment of dysplastic hip osteoarthrosis is necessary, osteotomy is preferable to fusion or THR. We evaluated periacetabular osteotomy as a method of choice. PATIENTS AND METHODS: We treated 36 symptomatic dysplastic hip joints (32 patients) with the Bernese periacetabular osteotomy (PAO) between 1994 and 2001. We used the ilio-inguinal (I-I) approach in 32 hips and a modified Smith-Petersen (S-P) approach in 4. The patients were followed for mean 4 (1.5-8) years. In 1 patient with coxa valga, a varus femoral osteotomy was performed 1 year after PAO. 2 hips, in which we used the modified S-P approach, necessitated a capsulotomy. RESULTS: The median Merle d'Aubigne score increased from 13 points preoperatively to 16 points postoperatively. This improvement in terms of pain, motion and ambulation was accompanied by spatial reorientation and correction. The lateral center edge angle of Wiberg (CE) improved from an average of 7 degrees to 28 degrees. The anterior center edge angle of Lequesne (FP) improved from an average of 18 degrees to 28 degrees. The acetabular index angle (AC) improved from an average of 22 degrees to 10 degrees. Major complications included 1 partial lesion of the sciatic nerve, 1 malunion and 1 combined nonunion of the pubic and ischiatic osteotomy. 2 patients underwent subsequent total hip replacement (THR) for progressive osteoarthrosis with pain. INTERPRETATION: We found good radiographic correction of deformities, improvement of hip function and pain relief with an acceptable complication rate. With appropriate patient selection, this procedure is the most physiological treatment of symptomatic hip dysplasia in young adults. In addition to relieving symptoms, it may prevent and postpone the development of secondary osteoarthrosis. PMID- 15788311 TI - Toxic serum gentamicin levels after the use of gentamicin-loaded sponges in infected total hip arthroplasty. AB - BACKGROUND: At our clinic, total hip arthroplasties with an acute postoperative infection are treated with surgical debridement and application of several gentamicin-loaded sponges, each with an equivalent of 130 mg gentamicin. We studied the possible systemic effect of this local application of gentamicin. PATIENTS AND METHODS: All 12 patients treated during 1997-2001 were included. 4-6 sponges were used in each case. We measured the serum gentamicin levels daily, from the first postoperative day until gentamicin was no longer measurable. The creatinine levels were monitored daily and the creatinine clearance was calculated according the Cockcroft-Gault formula. RESULTS: In 7 of the 12 patients, we found toxic serum levels of gentamicin (range 2-13 mg/L). In 3 cases there was a significant drop in renal clearance, which persisted. In 3 other cases there was a temporary decrease in renal clearance. INTERPRETATION: We found toxic serum levels after local administration of gentamicin. We even found a persistent decrease in renal clearance in some patients. Because of the alarming results, we have reduced the number of sponges used for this purpose. PMID- 15788312 TI - Comparison of analog and digital preoperative planning in total hip and knee arthroplasties. A prospective study of 173 hips and 65 total knees. AB - INTRODUCTION: Digital correction of the magnification factor is expected to yield more accurate and reliable preoperative plans. We hypothesized that digital templating would be more accurate than manual templating for total hip and knee arthroplasties. PATIENTS AND METHODS: Firstly, we established the interobserver and intraobserver reliability of the templating procedure. The accuracy and reliability of digital and analog plans were measured in a series of 238 interventions, which were all planned using both techniques. RESULTS: Interobserver reliability was good for the planning of knee arthroplasties (e values 0.63-0.75), but not more than moderate for the planning of hip arthroplasties (e-values 0.22-0.54). Analog plans of knee arthroplasties systematically underestimated the component sizes (1.1 size on average), while the digital procedure proved to be accurate (0.1-0.4 size too small on average). The following figures show percentage of cases receiving a correct implant, allowing an error of one size. Digital templating of the hip arthroplasty was less frequently correct (cemented cup and stem: 72% and 79%; uncemented cup and stem: 52% and 66%) than analog planning (cemented cup and stem: 73% and 89%; uncemented cup and stem: 64% and 52%). INTERPRETATION: Planning of component sizes for total knee arthroplasties is an accurate procedure when performed digitally. Our digital preoperative plans which were performed by someone other than the surgeon were less accurate than the analog plans prepared by the surgeon. PMID- 15788313 TI - Survival of the AGC total knee arthroplasty is similar for arthrosis and rheumatoid arthritis. Finnish Arthroplasty Register report on 8,467 operations carried out between 1985 and 1999. AB - We report the survival of AGC knee endoprosthesis from the Finnish Arthroplasty Register for 2 indications, osteoarthrosis (OA, 6,306 knees) and rheumatoid arthritis (RA, 2,161 knees) during 1985-1999. Survivorship analysis was performed with revision as an endpoint. We found similar survival rates. In the OA group, survival after 5 years was 97% and it was 94% after 10 years. In the RA group the corresponding figures were 97% and 96%, respectively. There was no significant difference in survival whether or not cement was used for fixation. The revision rates were higher in men and in younger patients. PMID- 15788314 TI - Bandaging technique after knee replacement. AB - BACKGROUND: Firm bandaging of the knee following knee replacement may prevent bleeding into the joint by a tamponade effect. We studied the pressure required to achieve tamponade, and then clinically compared the use of a compression bandage with the use of a standard crepe bandage, with or without a drain. METHOD: Transducers were used to measure the pressure achieved on the surface of the knee under different bandages, and within the knee following release of the tourniquet. We prospectively compared 3 series of 50 patients each: (1) with compression bandaging from toes to mid-thigh, (2) with crepe bandage from mid calf to mid-thigh alone, or (3) with crepe bandage and suction drain. RESULTS: The pressure within the joint at which tamponade occurs is 52-62 mm Hg. The pressure on the skin under a properly applied compression bandage is between 28 and 32 mm Hg, and this controls bleeding within the joint. Patients treated with compression bandaging recovered more quickly from the operation, had a shorter hospital stay, and a greater range of flexion on discharge. They had no swelling of the limb, rarely suffered a tense hemarthrosis, and had fewer complications. INTERPRETATION: The use of a compression bandage incorporating the foot and calf following knee replacement surgery, without the use of drains, confers specific advantages over the use of a crepe bandage alone. PMID- 15788315 TI - No beneficial effects of joint distraction on early microscopical changes in osteoarthrotic knees. A study in rabbits. AB - BACKGROUND: Although promising results have been reported, on the use of joint distraction in osteoarthrotic joints, the mechanism behind the effects has not yet been expland. MATERIAL AND METHODS: 24 rabbits were randomly divided into 4 groups and osteoarthrosis was induced by papain injection. The first group served as control and the others were treated by simple external fixation (group 2), articulated distraction (group 3) or nonarticulated distraction (group 4). RESULTS: Histologically, there was no significant difference between the first, the second and the third groups. However, osteoarthrosis increased in group 4. INTERPRETATION: We conclude that joint distraction has no beneficial effect on the osteoarthrotic cartilage in papain-induced osteoarthrosis, and nonarticulated distraction worsens the results. PMID- 15788316 TI - Compliance of Dutch orthopedic departments with national guidelines on thromboprophylaxis. A survey of Dutch orthopedic thromboprohylaxis. AB - All 110 Dutch orthopedic departments were sent a survey on perioperative thromboprophylaxis protocols, and 79% responded. After hip and knee replacements, all used pharmacological thromboprophylaxis: a low-molecular weight heparin (LMWH) in 87% of departments, which was most often combined with vitamin K antagonists (VKAs). LMWH was usually started preoperatively (91%). After discharge, VKAs were mostly prescribed (79%) for at least 6 weeks, and often for 3 months. 17% of departments used LMWH for 6 weeks, whereas in only 3% no post discharge prophylaxis was given. In day-care surgery, including arthroscopies, 58% use LMWH and in short-stay surgery 80% administer LMWH during the hospital stay. Because of lack of conclusive evidence for day-care surgery, the national guidelines cannot support pharmacological prophylaxis in this setting. In general, Dutch orthopedic departments comply poorly with the national guidelines on extended thromboprophylaxis for hip and knee replacement surgery, which recommends postoperative LMWH for 6 weeks. They are divided in the use of pharmacological prophylaxis in day-care surgery. PMID- 15788317 TI - Stable fixation of the ulnar component in the Kudo elbow prosthesis. A radiostereometric (RSA) study of 13 prostheses with 2-year follow-up. AB - BACKGROUND: Concern has been expressed about the large number of radiolucent lines around the ulnar component of the Kudo elbow prosthesis in medium-term follow-up. PATIENTS AND METHODS: We studied the metal-backed cemented ulnar component in 13 Kudo elbow prostheses (type 5) using radiostereometric analysis (RSA). All patients had rheumatoid arthritis. There were 2 men and 9 women with a mean age of 55 years. 2 were operated bilaterally. The metal-backed ulnar component was marked with three 0.8 mm tantalum spheres and the proximal ulna with 5 spheres of 0.8 or 1.0 mm diameter. The initial RSA examination was performed during the first week after the operation. Further examinations were done at 4, 12 and 24 months. Conventional radiographs were taken during the first week postoperatively, and at 12 and 24 months. RESULTS: Translations (medial/lateral, antero/posterior and proximal/distal) were less than 0.5 mm in all but 1 patient who had a maximal translation of 3.4 mm distally. The mean rotations around all three axes were less than 0.4 degrees. The patient who had a translation of 3.4 mm also had varus angulation exceeding 4 degrees. This patient also had progressive circumferential radiolucent lines on conventional radiographs. The Mayo elbow score increased from 40 (25-65) before surgery to 92 (45-100) at 2 years. INTERPRETATION: The fixation of the metal-backed ulnar component of the Kudo elbow prosthesis at 2 years is good. PMID- 15788318 TI - Pseudomonas aeruginosa biofilm formation and slime excretion on antibiotic-loaded bone cement. AB - BACKGROUND: Infection is an infrequent but serious complication of prosthetic joint surgery. These infections will usually not clear until the implant is removed and re-implantation has a high failure rate, especially when Pseudomonas aeruginosa is involved. MATERIAL AND METHODS: We examined Pseudomonas aeruginosa biofilm formation on plain and gentamicin-loaded bone cement with confocal scanning laser microscopy (CSLM). Two different stains were applied in order to visualize and quantify the distribution of bacterial cells and extracellular polymeric substances (slime) from the bone cement surface to the top of the biofilm. Staining with LIVE/DEAD viability stain differentiated between live and dead bacteria within the biofilm, and slime production was evaluated after staining with Calcofluor white. RESULTS: CSLM showed that the biofilm was a nonuniform structure of variable thickness, with differences in local bacterial cell and slime densities. Incorporation of gentamicin in bone cement resulted in a 44% reduction in bacterial viability, while the slime density increased significantly. In addition, conventional plate counting showed the development of small-colony variants on gentamicin-loaded bone cement with a decreased sensitivity for gentamicin (MIC: 8 m/L), as compared with normal-sized colonies taken from plain and gentamicin-loaded bone cement (MIC: 3 m/L). The enhanced slime production on antibiotic-loaded bone cement, together with the formation of small-colony variants, resulted in decreased susceptibility to antibiotics- probably concomitant with the onset of persistent and relapsing infections. INTERPRETATION: In the clinical situation, our findings help to explain the frequent re-implantation failure of joint replacements infected with P. aeruginosa when the procedure has been performed using antibiotic-loaded bone cement. PMID- 15788320 TI - Biodegradable X-ray markers of controlled radio-opacity. Temporary position measurements in bone. AB - In order to analyze X-ray markers for potential use in biodegradable implants or radiostereogrammatic analysis (RSA), we combined iopromide contrast fluid with biodegradable calcium phosphate cement. The radio-opacity of 10 x 10 mm markers containing different iodine concentrations (0, 120, 240, 360 and 720 mg per gram cement) was compared to an aluminium wedge of increasing (1-10 mm) thickness. The addition of iopromide increased the radio-opacity in a dose-dependent manner, which was comparable to 9-mm aluminium at concentrations of 240-720 mg/g. Radiographs of markers placed in explanted rabbit and in human femora were made to investigate the clinical accuracy for position determination. Markers of 1 x 1 mm (120 mg/g) were clearly discernable in all femora, and could be used to adequately measure distances of 5-45 mm (accuracy 0.10-2.19 mm). These markers might be embedded in biodegradable implants or used as temporary markers in the bone to analyze postoperative position on radiographs. PMID- 15788319 TI - Cytokine profiles in conditioned media from cultured human intervertebral disc tissue. Implications of their effect on bone marrow stem cell metabolism. AB - BACKGROUND: Cytokines released from intervertebral discs cultured in vitro have not been profiled, and the effect of these cytokines on human bone marrow stem cells is yet to be studied. MATERIALS AND METHODS: Intervertebral discs from 14 patients who had undergone spinal fusion surgery were cultured separately in vitro. Conditioned media were collected after 48 and 96 h of culture in serum free Minimum Essential Medium (MEM). Profiling of the cytokines was conducted using pooled media. Conditioned medium from each patient was also tested in human bone marrow stem cell culture, and incorporation of alkaline phosphatase and 3H thymidine incorporation was evaluated. RESULTS: Of the 18 cytokines screened, 12 were found to be positive, but only eotaxin, IP-10, Rantes IL-6 and IL-8 seemed to be present at high levels. There was a close correlation between IL-6 and IL-8 levels in the medium (R = 0.90, p < 0.001). When the conditioned media were added to human bone marrow stem cell cultures, cellular proliferation was stimulated (p = 0.02), but alkaline phosphatase activity remained unchanged. Cellular proliferation correlated negatively with IL-6 levels (R = -0.44, p = 0.04). INTERPRETATION: Intervertebral discs secrete certain cytokines into the medium when cultured in vitro, and conditioned media from cultured intervertebral discs stimulate proliferation of bone marrow stem cells. PMID- 15788321 TI - Adductor tenotomy in spastic cerebral palsy. A long-term follow-up study of 78 patients. AB - BACKGROUND: There is a risk of hip dislocation in children with spastic cerebral palsy. We evaluated the prophylactic effect of adductor tenotomy in patients with long-term follow-up. PATIENTS AND METHODS: Our material comprised 78 patients (46 boys) with a mean age of 8 (2-17) years who underwent adductor tenotomy during the period 1986-1991. 40 patients had spastic diplegia and 38 had quadriplegia. For patients who had further hip surgery, follow-up was until the next hip operation. Those who had not undergone further surgery were invited to a follow up examination. The migration percentage (MP) was measured on the preoperative and follow-up radiographs. The radiographic result was termed good if MP at follow-up was reduced or had increased less than 10%. The follow-up period was 10 (1.6-16) years, with a mean of 6 years for patients with later hip surgery and 13 years without such surgery. RESULTS: The clinical outcome was good in 51 cases, poor in 12, and uncertain in 15. The radiographic result was good in 39 of the 53 patients with radiographs available both preoperatively and at follow-up. The patients with good radiographic results had lower preoperative MP than those with poor results (MP 34% versus 49%) and lower preoperative acetabular index. The mean increase in MP (worst hip in each patient) was 1.9% per year, which is considerably less than that in nonoperated patients. Further hip surgery was necessary in 27 patients, because of increasing MP in 14 cases and for clinical reasons in 13. INTERPRETATION: Adductor tenotomy reduced the trend towards lateral displacement of the hip joints. The operation had a favorable outcome in approximately two-thirds of the patients. The operation should be performed before the MP reaches 50%. PMID- 15788322 TI - A bleeding pseudoaneurysm of the lateral genicular artery after total knee arthroplasty--a case report. PMID- 15788323 TI - Changes in load-bearing area after Ganz periacetabular osteotomy. PMID- 15788326 TI - Perceptions of participation and predictors of perceived problems with participation in persons with spinal cord injury. AB - OBJECTIVE: To describe how persons with spinal cord injury perceived their participation in life situations and to determine the relationship between their participation and perceived problems therewith. The purpose was also to evaluate the influence of age, sex, level of injury, time since injury, marital status and access to social support on perceived problems with participation. DESIGN: Cross sectional. SUBJECTS: One hundred sixty-one persons with spinal cord injury. METHODS: A postal questionnaire including socio-demographic characteristics and a Swedish version of the Impact on Participation and Autonomy questionnaire. RESULTS: A majority of the respondents perceived their participation as sufficient in most activities addressed. Still, a majority of the respondents perceived one or more severe problems with their participation. Access to social support was the most influencing variable in predicting perceived severe problems with participation as compared with certain personal and health-related factors. CONCLUSION: The results suggest that it is important to consider access to social support along with other factors in the person-environment interaction and their influence on severe problems with participation in enhancing clients' participation in rehabilitation. PMID- 15788327 TI - Relations between coping strategies and health-related quality of life in patients with spinal cord lesion. AB - OBJECTIVE: Although the use of appropriate coping strategies has been suggested to be a key factor in determining successful adjustment to severe physical illness/disability, little systematic support for this link has been found. We investigated relationships between spinal cord lesion-related coping strategies and health-related quality of life when studying for sociodemographic, disability related and social support variables. DESIGN AND SUBJECTS: We studied 256 persons with traumatically acquired spinal cord lesion (=1 year) from a typical rural/urban Swedish area in a cross-sectional design. METHODS: Coping measure was the Spinal Cord Lesion-related Coping Strategies Questionnaire. Outcome measures were the Spinal Cord Injury Quality of Life Questionnaire, the Short-Form 36 Health Survey version 2.0, and a standardized global question of overall quality of life. Multiple regressions were performed. RESULTS: Coping strategies were clear correlates of health-related quality of life when sociodemographic, disability-related and social support variables were studied. The relationship between coping strategies and quality of life was: the more revaluation of life values (Acceptance) and the fewer tendencies towards dependent behaviour (Social reliance) the better the health-related quality of life. CONCLUSION: Our results suggest that greater focus needs to be directed to coping strategies and to ways of facilitating adaptive outcomes in rehabilitation. PMID- 15788328 TI - Factors determining job retention and return to work for disabled employees: a questionnaire study of opinions of disabled people's organizations in the UK. AB - OBJECTIVES: To determine the views of organizations of and for disabled people in order to inform the writing of the British Society of Research Medicines policy document "Vocational Rehabilitation--The Way Forward". PATIENTS/ORGANIZATIONS: A single mailing was sent to 98 disability organizations within the UK. DESIGN: A semi-structured postal questionnaire focused on factors (i) within the National Health Service; (ii) external to it, mainly in the workplace, making it difficult for people to stay in work in the presence of disease/disability, or to find work after losing their job (within the last 6 months). RESULTS: A 30% response rate, with many incomplete questionnaires, was obtained so that 24 complete questionnaires were analysed. The dominant findings concerning the National Health Service were, overwhelmingly, that it was perceived as impacting deleteriously on the work of disabled people with delays to consultation, investigation and rehabilitation and a lack of appreciation of workplace issues. Employers were seen as unresponsive to the needs of workers, with negative attitudes to disability. The changes required in both areas were closely related to these findings. CONCLUSION: Though the organizations surveyed were not representative, nevertheless there was considerable agreement about the need for both the National Health Service and employers to be more responsive to the workplace needs of disabled people. PMID- 15788329 TI - Cross-cultural validity of functional independence measure items in stroke: a study using Rasch analysis. AB - OBJECTIVE: To analyse cross-cultural validity of the Functional Independence Measure (FIM) in patients with stroke using the Rasch model. SETTINGS: Thirty-one rehabilitation facilities within 6 different countries in Europe. PARTICIPANTS: A total of 2546 in-patients at admission, median age 63 years. METHODS: Data from the FIM were evaluated with the Rasch model, using the Rasch analysis package RUMM2020. A detailed analysis of scoring functions of the 7 categories of the FIM items was undertaken prior to testing fit to the model. Categories were re-scored where necessary. Analysis of Differential Item Functioning was undertaken in pooled data for each of the FIM motor and social-cognitive scales, respectively. RESULTS: Disordered thresholds were found on most items when using 7 categories. Fit to the Rasch model varied between countries. Differential Item Functioning was found by country for most items. Adequate fit to the Rasch model was achieved when items were treated as unique for each country and after a few country specific items were removed. CONCLUSION: Clinical collected data from FIM for patients with stroke cannot be pooled in its raw form, or compared across countries. Comparisons can be made after adjusting for country-specific Differential Item Functioning, though the adjustments for Differential Item Functioning and rating scales may not generalize to other samples. PMID- 15788330 TI - Controlled study of neuroprosthetic functional electrical stimulation in sub acute post-stroke rehabilitation. AB - OBJECTIVE: Assess the effects of daily neuroprosthetic (NESS Handmaster) functional electrical stimulation in sub-acute stroke. DESIGN: Controlled study, patients clinically stratified to 2 groups; no active finger movement, and partial active finger movements, and then randomized to control and neuroprosthesis groups. Observer blinded evaluations at baseline and completion of the 6-week study. SUBJECTS: 22 patients with moderate to severe upper limb paresis 3-6 months post-onset. METHODS: Patients in day hospital rehabilitation, receiving physical and occupational therapy 3 times weekly. The neuroprosthesis group used the device at home. RESULTS: The neuroprosthesis group had significantly greater improvements in spasticity, active range of motion and scores on the functional hand tests (those with partial active motion). Of the few patients with pain and oedema, there was improvement only among those in the neuroprosthesis group. There were no adverse reactions. CONCLUSION: Supplementing standard outpatient rehabilitation with daily home neuroprosthetic activation improves upper limb outcomes. PMID- 15788332 TI - Clinical and administrative outcomes during publicly-funded inpatient stroke rehabilitation based on a case-mix group classification model. AB - OBJECTIVES: To determine efficiency and efficacy of publicly-funded inpatient stroke rehabilitation based on a Case-Mix Group Classification Model, and to analyse the usefulness of this decisional aid in the refinement of rehabilitation services delivery needed to optimize accessibility to inpatient rehabilitation services for individuals with stroke in a publicly-funded healthcare system. DESIGN: Individuals with stroke (n=422) who received inpatient rehabilitation through the Montreal Rehabilitation Hospital Network were included in this retrospective study. Clinical (total, motor and cognitive-Functional Independence Measure (FIM) scores, percentage of discharge to community) and administrative outcomes (onset to rehabilitation interval, length of inpatient rehabilitation stay, length of stay efficiency) were measured. RESULTS: Across Case-Mix Groups, mean onset to rehabilitation days varied between 16.2 (5.7) and 32.0 (19.4) days whereas the mean length of stay fluctuated between 27.5 (13) and 77.0 (27) days. Best total (41.6 (21.4)) and motor-FIM (38.9 (19.0)) gains were observed in most severely disabled cases (114) whereas the Case-Mix Group 103 presented the best cognitive-FIM gain (5.8 (4.0)). Optimal mean total, motor and cognitive-FIM efficiency rates, found in moderately disabled stroke patients, were 0.668 (0.434), 0.634 (0.377) and 0.15 (0.136), respectively. Majority of patients returned home following rehabilitation in all Case-Mix Groups (63.6% to 96.4%) except for groups 112 and 108. CONCLUSION: Moderate to good length of stay efficiencies are observed among all Case-Mix Group following stroke rehabilitation. In fact, individuals with moderate disability present the best rate of recovery. Variations in length of stay efficiency suggest that the use of a Case-Mix Group Classification Model in stroke rehabilitation could represent an innovative approach, especially for program evaluation in publicly-funded and universal-access rehabilitation hospitals. PMID- 15788331 TI - Development of the home and community environment (HACE) instrument. AB - OBJECTIVE: To develop and pilot test the Home and Community Environment instrument (HACE), a self-report measure designed to characterize factors in a person's home and community environment that may influence level of participation. DESIGN: A cross-sectional survey. SUBJECTS: Sixty-two adults recruited from community organizations and an outpatient rehabilitation center. METHODS: Six environmental domains were assessed: (i) home mobility; (ii) community mobility; (iii) basic mobility devices; (iv) communication devices; (v) transportation factors; and (vi) attitudes. Descriptive statistics, Kappa statistics and Kruskal-Wallis tests were used to ascertain whether persons were capable of assessing characteristics of their environment, could do so reliably and whether the distribution of environmental factors differed by type of living situation. RESULTS: Participants were capable of characterizing their home environment and most aspects of their community with acceptable reliability. The median percent agreement of the 6 environmental domains ranged from 75% to 100% (median Kappa values ranged from 0.47 to 1.0). Percent agreement for individual HACE items ranged from 58% to 100%. The lowest reliability values were observed in the community mobility domain. As hypothesized, individuals who lived in private homes characterized home and community mobility factors differently from those who lived in multi-unit complexes; evidence of HACE's validity. CONCLUSION: HACE is a promising self-report instrument for assessing characteristics of an individual's home and community environments. Additional research is needed to assess its utility for rehabilitation research. PMID- 15788333 TI - S100 and cognitive impairment after mild traumatic brain injury. AB - OBJECTIVE: The aim of this study was to explore the relationship between the proteins S100B and S100A1B and symptoms and signs of cognitive impairment for 3 months after mild traumatic brain injury (MTBI). METHODS: Serum concentrations of S100A1B and S100B were examined in a prospective cohort study of patients with MTBI and a Glasgow Coma Scale score of 14 or 15. Cognitive performance was assessed by repeated computerized neuropsychological testing and an extended neuropsychological test. Symptoms were assessed using the Rivermead Post Concussion Symptoms Questionnaire. RESULTS: Concentrations of S100B and S100A1B were above cut-off in 31% and 48% respectively. Eight percent of the patients had signs of cognitive impairment according to the computerized neuropsychological tests and 30% according to the extended test. Symptoms of cognitive impairment were reported by 44% of the patients on the first day post-injury and by 26% at 3 months. No significant associations between S100B or S100A1B concentrations and symptoms or signs of cognitive impairment were found. CONCLUSION: Abnormal S100 serum concentrations and symptoms or signs of cognitive impairment were not significantly associated in patients with MTBI and a Glasgow Coma Scale score of 14 or 15. PMID- 15788334 TI - Comparison of self-management methods for osteoarthritis and rheumatoid arthritis. AB - OBJECTIVE: To determine and compare self-management methods used for osteoarthritis and rheumatoid arthritis and to determine which methods patients consider most effective. METHODS: Patients with osteoarthritis and rheumatoid arthritis were questioned about self-management methods and asked to indicate the most effective ones. RESULTS: Twenty-seven patients with osteoarthritis and 26 with rheumatoid arthritis were interviewed. Exercise, joint protection, assistive devices and heat were the methods most commonly used. More patients with osteoarthritis than with rheumatoid arthritis used assistive devices (p=0.042). Exercise, assistive devices and heat were considered the most effective self management methods. More patients with osteoarthritis than with rheumatoid arthritis considered assistive devices to be the most effective method (p=0.016). CONCLUSION: More patients with osteoarthritis than with rheumatoid arthritis used and indicated assistive devices as the most effective self-management method. Similar numbers of patients in both groups used other self-management methods. PMID- 15788335 TI - Comments on the task force report on mild traumatic brain injury: journal of rehabilitation medicine supplement 43. PMID- 15788340 TI - Relation between stimulation characteristics and clinical outcome in studies using electrical stimulation to improve motor control of the upper extremity in stroke. AB - OBJECTIVE: Electrical stimulation can be applied in a variety of ways to the hemiparetic upper extremity following stroke. The aim of this review is to explore the relationship between characteristics of stimulation and the effect of electrical stimulation on the recovery of upper limb motor control following stroke. METHODS: A systematic literature search was performed to identify clinical trials evaluating the effect of electrical stimulation on motor control. The reported outcomes were examined to identify a possible relationship between the reported effect and the following characteristics: duration of stimulation, method of stimulation, setting of stimulation parameters, target muscles and stage after stroke. RESULTS: Nineteen clinical trials were included, and the results of 22 patient groups were evaluated. A positive effect of electrical stimulation was reported for 13 patient groups. Positive results were more common when electrical stimulation was triggered by voluntary movement rather than when non-triggered electrical stimulation was used. There was no relation between the effect of electrical stimulation and the other characteristics examined. CONCLUSION: Triggered electrical stimulation may be more effective than non triggered electrical stimulation in facilitating upper extremity motor recovery following stroke. It appears that the specific stimulus parameters may not be crucial in determining the effect of electrical stimulation. PMID- 15788341 TI - Reliability of gait performance tests in men and women with hemiparesis after stroke. AB - OBJECTIVE: To assess the reliability of 6 gait performance tests in individuals with chronic mild to moderate post-stroke hemiparesis. DESIGN: An intra-rater (between occasions) test-retest reliability study. SUBJECTS: Fifty men and women (mean age 58+/-6.4 years) 6-46 months post-stroke. METHODS: The Timed "Up & Go" test, the Comfortable and the Fast Gait Speed tests, the Stair Climbing ascend and descend tests and the 6-Minute Walk test were assessed 7 days apart. Reliability was evaluated with the intraclass correlation coefficient (ICC(2,1)), the Bland & Altman analysis, the standard error of measurement (SEM and SEM%) and the smallest real difference (SRD and SRD%). RESULTS: Test-retest agreements were high (ICC(2,1) 0.94-0.99) with no discernible systematic differences between the tests. The standard error of measurement (SEM%), representing the smallest change that indicates a real (clinical) improvement for a group of individuals, was small (< 9%). The smallest real difference (SRD%), representing the smallest change that indicates a real (clinical) improvement for a single individual, was also small (13-23%). CONCLUSION: These commonly used gait performance tests are highly reliable and can be recommended to evaluate improvements in various aspects of gait performance in individuals with chronic mild to moderate hemiparesis after stroke. PMID- 15788342 TI - Speed and physiological cost index of hemiplegic patients pedalling a wheelchair with both legs. AB - OBJECTIVE: To determine whether hemiplegic patients can propel a leg-pedalling wheelchair and whether it is easier and faster for them to pedal the wheelchair using both legs alternately than to propel an ordinary wheelchair with their unaffected hand and leg. DESIGN: Within-subject comparison. SUBJECTS: Subjects comprised 10 hemiplegic in-patients (8 males, 2 females), aged 63.7 (SD 12.7) years with severe or moderate gait disturbance due to stroke. METHODS: Subjects were asked to practice propelling the leg-pedalling wheelchair and ordinary wheelchair on both slalom and rectangular courses for a period of 7-10 days. Once they had become skilled in this, the wheelchair speed and patient's heart rate were measured, and a physiological cost index was calculated. RESULTS: Subjects could pedal the leg-pedalling wheelchair using both legs alternately. The speed of this wheelchair was faster than that of the ordinary wheelchair, and the physiological cost index for pedalling it was lower than that for propelling the ordinary wheelchair. However, subjects needed some help in transferring to the leg-pedalling wheelchair. CONCLUSION: The hemiplegic patients could pedal the leg pedalling wheelchair using both legs alternately faster and more effectively with regard to speed and physiological cost index. PMID- 15788343 TI - Use of analgesic drugs in individuals with spinal cord injury. AB - OBJECTIVE: This study set out to elucidate which factors are associated with or predictive for the use of analgesic drugs in patients with spinal cord injury and pain. DESIGN: A cross-sectional descriptive study with a partly prospective cohort. PATIENTS: One hundred and twenty-three patients with a spinal cord injury matched for gender, age, level of lesion and completeness of injury. METHODS: Questionnaires consisting of ratings in the areas of pain intensities, pain unpleasantness, life satisfaction, anxiety and depression, and questions about consumption of analgesic drugs were posted to the 123 patients. RESULTS: Of the 101 patients (82.1%) who returned the questionnaire, 90 (46 women and 44 men) still suffered from pain and were thus included in the study. Statistical analysis showed that although the number of pain medications used per person had increased in the last 3 years, the ratings of pain were unaffected. Logistic regression analyses also revealed that the use of pain-relieving medication was associated with higher ratings on the affective component of pain, lower ratings of leisure activities and the presence of stabbing/cutting pain. CONCLUSION: The affective component of pain is the main predictor for the use of analgesics in patients with a spinal cord injury. Complementary strategies, including a multidisciplinary approach, for relieving the unpleasantness of pain need to be explored further. PMID- 15788344 TI - Disability, pain, psychological factors and physical performance in healthy controls, patients with sub-acute and chronic low back pain: a case-control study. AB - OBJECTIVE: To compare measures of disability, psychological factors, pain and physical performance in healthy controls, and patients with sub-acute and chronic low back pain. To evaluate the concept of the deconditioning syndrome and to explore factors that may contribute to chronicity. DESIGN: Case-control study. SUBJECTS: Three age- and gender-matched groups were included in the study; healthy controls (n = 45), patients sick-listed 8-12 weeks (n = 46) and patients with chronic low back pain on a waiting list for lumbar instrumented fusion (n = 45). METHODS: Measures of disability, pain, psychological factors, and physical performance were obtained from the 3 groups using validated measures. RESULTS: Gender, age, body weight and height were not significantly different between the groups. Comparable scores were found for self-rated working ability, fear avoidance beliefs for physical activity and aerobic capacity in the 2 patient groups. Oswestry Disability Index, pain, emotional distress, abdominal and back muscle endurance were significantly different between the 3 groups. Self-efficacy for pain and fear-avoidance beliefs for work was significantly different between the 2 patient groups. CONCLUSION: The results suggest a stepwise deterioration of impairment and disability from healthy controls to patients with chronic low back pain. Most variables distinguished between healthy controls and patients with sub acute or chronic low back pain. Deconditioning was more related to psychophysical measures of abdominal and back muscle endurance than to cardiovascular fitness. Comparable scores for fear-avoidance and working ability in the 2 patient categories suggest that these factors appear at an early stage and contribute to the transition from acute to chronic low back pain. PMID- 15788345 TI - Coping strategy use: does it predict adjustment to chronic back pain after controlling for catastrophic thinking and self-efficacy for pain control? AB - OBJECTIVE: To explore whether coping strategy use predicted levels of adjustment in chronic low back pain after controlling for the influence of catastrophic thinking and self-efficacy for pain control. METHODS: Eighty-four patients with chronic low back pain completed the Coping Strategies Questionnaire, a pain VAS and the Roland Disability Questionnaire. To derive composite measures of coping, the Coping Strategies Questionnaire subscales, excluding the Catastrophizing subscale and 2 single-item scales, were entered into a principal components analysis. The extent to which scores on the coping measures predicted levels of adjustment after controlling for catastrophic thinking (Catastrophizing subscale) and self-efficacy for pain control (2 single-item scales) was explored using sequential multiple regression analysis. RESULTS: Two coping dimensions emerged from the principal components analysis, which were labelled Distraction and Praying or Hoping, and Denial of Pain and Persistence. Scores obtained on these coping measures explained an additional 5% and 13% of the variance in pain intensity and disability, respectively. Interestingly, however, the scores on the coping measures did not predict pain intensity or disability after controlling for the influence of catastrophic thinking and self-efficacy for pain control. CONCLUSION: Coping strategy use might only be related to levels of adjustment via the effect it has on catastrophic thinking and self-efficacy for pain control. PMID- 15788346 TI - Correlation between a novel upper limb activity monitor and four other instruments to determine functioning in upper limb complex regional pain syndrome type I. AB - OBJECTIVE: To determine the place of a novel Upper Limb Activity Monitor in the field of instruments measuring functioning and health in upper limb complex regional pain syndrome type I, by exploring the correlation between the Upper Limb Activity Monitor and 4 questionnaires. METHOD: Subjects (n = 30) were measured at home and correlations were calculated between the Upper Limb Activity Monitor and 4 questionnaires; Sickness Impact Profile, RAND-36 Health Survey, Disabilities of Arm Shoulder Hand Questionnaire and Radboud Skills Questionnaire. RESULTS: Of the inter-questionnaire correlations 83% were significant, whereas 46% of the correlations between the Upper Limb Activity Monitor and the questionnaires were significant. The number and strength of the correlations between the Upper Limb Activity Monitor and questionnaires was dependent on the degree to which similar aspects of functioning were measured. CONCLUSION: The Upper Limb Activity Monitor has some correlation with other instruments related to functioning and health, but generally it does not measure the same areas. PMID- 15788348 TI - Fever during post-acute rehabilitation in patients with brain injury. AB - OBJECTIVE: To investigate the incidence of fever during the post-acute rehabilitation phase in patients with brain injury. DESIGN: Prospective study. SUBJECTS: Seventy-four patients with brain injury. METHODS: Patients were evaluated for the presence of fever during the rehabilitation phase. Demographics, time since injury, acute hospital and rehabilitation stay, brain injury and clinical characteristics were recorded for all subjects. RESULTS: Of the study group, 36 patients (36/74; 48.6%) had at least 1 event qualified as fever. The most common cause of fever was urinary tract infection. There were only 2 patients considered as having central fever. Fever was significantly more frequent in patients with a greater number of neurological impairments, more severe neurological impairments and a lower level of independence. The rehabilitation stay was prolonged in patients with fever. The average length of rehabilitation stay was 62.5 days and 49.8 days in patients with or without fever, respectively. CONCLUSION: Infection is the most common cause of fever after brain injury and the incidence of central fever is low. PMID- 15788347 TI - Exercise therapy for shoulder pain aimed at restoring neuromuscular control: a randomized comparative clinical trial. AB - OBJECTIVE: To compare the effectiveness of exercise therapy aimed at restoring neuromuscular control mechanisms at the shoulder with other conservative interventions for the treatment of chronic shoulder pain with and without accompanying stiffness. DESIGN: Randomized control trial. PATIENTS: A total of 138 volunteers with unilateral shoulder pain of local mechanical origin. METHODS: Subjects were randomly allocated to receive exercise therapy aimed at restoring dynamic stabilizing mechanisms and muscle co-ordination at the shoulder; or subacromial corticosteroid injection; or a combination of physical modalities and range of motion exercises. Outcome measurements of pain intensity, functional impairment, active range of motion, isometric muscle force and self-assessed improvement were taken at baseline and after 5 weeks of treatment. RESULTS: The mean/median changes in all outcome measurements at 5 weeks indicated that subjects in each treatment group, improved significantly with no difference between the treatment groups. Comparison with an earlier no-treatment trial would suggest that this improvement is greater than that which could be expected by natural recovery. CONCLUSION: Exercise therapy aimed at restoring neuromuscular control, corticosteroid injection and multiple physical modalities and range of motion exercises are equally effective in the short-term treatment of shoulder pain, with exercise therapy and corticosteroid injection being less costly to administer. PMID- 15788351 TI - Growth patterns of Filipino children indicate potential compensatory growth. AB - BACKGROUND: The extent to which compensatory growth can occur after 2 years of age among children who were stunted in infancy has been questioned due, in part, to limitations in the data used to explore the issue. AIM: This study uses longitudinal data with multiple measurements over the entirety of the growth period to explore the potential for post-infancy compensatory growth in a developing country context. SUBJECTS AND METHODS: Data comes from >2000 Filipino youths from the Cebu Longitudinal Health and Nutrition Survey. Anthropometric data were collected bimonthly from 0 to 2 years, and at mean ages of 8, 11.5, 15.5 and 18.5 years. Growth indicators and influences on growth are compared among three groups: those that improved, tracked or worsened in change in height for-age Z-score from 2 to 18.5 years of age. RESULTS: Those that improved in growth from 2 to 18.5 years grew slowest before 2 years of age, while the opposite was true among those that worsened. The reversal took place during the prepubescent growth spurt. Those that improved tended to have less educated but taller mothers. Regression to the mean and the potential for compensatory growth are discussed. PMID- 15788352 TI - Growth of Chinese Italian infants in the first 2 years of life. AB - BACKGROUND: Numerous studies have been carried out on the growth of human migrant populations. However, studies on the auxological pattern of Chinese migrant population are limited in Italy. AIMS: In this paper we examine the growth pattern from birth to 24 months and the body composition of Chinese infants born and living in Bologna (north Italy). SUBJECTS AND METHODS: A longitudinal study was conducted recruiting 224 healthy Chinese children. The anthropometric measurements were collected when the children were immunized. Body mass index (BMI), Arm Muscle Area and Arm Fat Area were calculated. RESULTS: The growth of Chinese Italian children is adequate, being within the normal limits according to the NCHS reference standard. In comparison with Chinese children living in China and abroad, their body length is greater. Comparisons with growth curves for Italian children (Emilia-Romagna) show that weight and length values are higher in Chinese children than in Italians until 12 months, and comparable thereafter. The weight and length growth velocities are similar to those reported for Italian children. CONCLUSION: As a whole, the results of the present study support the hypothesis that Chinese children born and living in Italy grow in an appropriate environment to achieve their growth potential. PMID- 15788353 TI - A study of the relationships between angiotensin- converting enzyme gene, chymase gene polymorphisms, pharmacological treatment with ACE inhibitor and regression of left ventricular hypertrophy in essential hypertension patients treated with benazepril. AB - BACKGROUND: About 15-37% of the adult population worldwide suffers from hypertension. Hypertension is responsible for one-third of all global deaths. Left ventricular hypertrophy (LVH) is one of the most important characteristics of hypertension target organ damage and is also an independent risk factor for cardiovascular events. Therefore, effective regression of LVH is a main aim of hypertension treatment and also an important public health concern. However, few studies of the regression of LVH have been reported. In particular, little is known about the relationship between the genotypes of angiotensin-converting enzyme (ACE) and chymase (CMA) genes, and the effectiveness of antihypertensive drugs in regression of LVH. AIM: The study investigated whether the insertion/deletion (I/D) polymorphism of ACE gene and the A/B polymorphism of the CMA gene are related to the regression of LVH in essential hypertension patients who were participants in a long-term trial of therapy with benazepril. SUBJECTS AND METHODS: Data from 157 patients was collected and used in the analysis. The genotypes of ACE and CMA genes were identified by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Left ventricular mass (LVM) was measured by echocardiography, and left ventricular mass index (LVMI) was calculated. RESULTS: Blood pressure was markedly reduced and heart rate was unchanged by long-term treatment with benazepril. Regression of LVH was observed. The mean reduction in LVMI was 41.50+/-28.48 g m-2. Reduction of LVM, LVMI and percentage reduction of LVMI were more in the DD group than in the II and ID groups (P<0.05). No significant difference in other indices was found in the different genotype groups of ACE (P>0.05). No significant difference in all indices was found among the different genotype groups of CMA (P>0.05). No interaction was found between the genotypes of ACE and CMA. CONCLUSION: Hypertension patients with the DD genotype are more likely to have regression of LVH when treated with benazepril than patients with other genotypes of ACE. No evidence was found to support an association between CMA genotype and regression of LVH in patients or to support the interaction between the two genes in regression of LVH. PMID- 15788354 TI - Sexual dimorphism in foot length proportionate to stature. AB - BACKGROUND: The preponderance of existing results suggests that, relative to stature, women have smaller feet than men. However, several investigations indicate that the relationship between foot length and stature may be curvilinear, a pattern that, due to the dimorphic nature of stature, would mask the true direction of pedal sexual dimorphism in published results. AIM: The study aimed to determine whether proportionate foot length is sexually dimorphic and, if so, the nature of that dimorphism. MATERIALS AND METHODS: Surveying genetically disparate populations (USA, Turkey, and Native North and Central American), we examined data from three previous anthropometric studies (Davis 1990, Parham et al. 1992, Ozaslan et al. 2003) and foot tracings from the Steggerda Collection at the US National Museum of Health and Medicine. Analyses explored sex differences in the ratio between foot length and stature, and tested for nonlinearity. RESULTS: Although varying in degree across populations, proportionate to stature, female foot length is consistently smaller than male foot length. CONCLUSION: Given the biomechanical challenges posed by pregnancy, smaller female proportionate foot length is somewhat surprising, as foot length affects dorsoventral stability. It is possible that the observed pattern reflects intersexual selection for small female foot size, a cue of youth and nulliparity. PMID- 15788355 TI - Validity of BMI as a measure of obesity in Australian white Caucasian and Australian Sri Lankan children. AB - BACKGROUND: Body mass index (BMI) is used to diagnose obesity. However, its ability to predict the percentage fat mass (%FM) reliably is doubtful. Therefore validity of BMI as a diagnostic tool of obesity is questioned. AIM: This study is focused on determining the ability of BMI-based cut-off values in diagnosing obesity among Australian children of white Caucasian and Sri Lankan origin. SUBJECTS AND METHODS: Height and weight was measured and BMI (W/H2) calculated. Total body water was determined by deuterium dilution technique and fat free mass and hence fat mass derived using age- and gender-specific constants. A %FM of 30% for girls and 20% for boys was considered as the criterion cut-off level for obesity. BMI-based obesity cut-offs described by the International Obesity Task Force (IOTF), CDC/NCHS centile charts and BMI-Z were validated against the criterion method. RESULTS: There were 96 white Caucasian and 42 Sri Lankan children. Of the white Caucasians, 19 (36%) girls and 29 (66%) boys, and of the Sri Lankans 7 (46%) girls and 16 (63%) boys, were obese based on %FM. The FM and BMI were closely associated in both Caucasians (r=0.81, P<0.001) and Sri Lankans (r=0.92, P<0.001). Percentage FM and BMI also had a lower but significant association. Obesity cut-off values recommended by IOTF failed to detect a single case of obesity in either group. However, NCHS and BMI-Z cut-offs detected cases of obesity with low sensitivity. CONCLUSIONS: BMI is a poor indicator of percentage fat and the commonly used cut-off values were not sensitive enough to detect cases of childhood obesity in this study. In order to improve the diagnosis of obesity, either BMI cut-off values should be revised to increase the sensitivity or the possibility of using other indirect methods of estimating the %FM should be explored. PMID- 15788356 TI - Childhood obesity in United Arab Emirates schoolchildren: a national study. AB - OBJECTIVE: The study assessed the national prevalence of obesity among schoolchildren in the United Arab Emirates (UAE). SUBJECTS: A stratified 10% random sample of 16,391 children was drawn from 145,492 pupils in the UAE. METHODS: Height and weight were measured by physicians and trained nurses, and the body mass index (BMI; weight (kg)/height (m)2) was computed. BMIs for UAE were compared to recently published international standards. RESULTS: Comparison of BMIs to international reference data revealed that UAE children are at increased risk for overweight (>25 kg m-2 and =30 kg m-2) and obesity (=30 kg m 2). For example, 10-year-old male UAE children had 1.7 times the rate of overweight compared to international standards and 1.9 times at 18 years. Similarly, female UAE children have 1.8 times the rate of overweight compared to international standards at 10 and 18 years of age. Obesity was 2.3-fold higher among UAE males at 14 years compared to international standards, and increased to 3.6 times at 18 years of age. Among UAE female children, obesity was same as males at 14 years, 2.3 times than the international standards. At 18 years of age, UAE female obesity was 1.9-fold higher than the international standard, nearly one-half the rate of obesity among UAE males at the same age. CONCLUSION: The frequency of obesity among UAE youth is two to three times greater than the recently published international standard. Profound public health implications of childhood obesity for UAE children and young adults are seriously increased because of adult chronic disease processes (e.g. cardiovascular disease, diabetes) normally attendant to increased obesity rates. PMID- 15788357 TI - The distribution of Y chromosome haplogroups in the nationalities from Yunnan Province of China. AB - The genetic structure of 26 identified nationalities from Yunnan Province of China was studied using Y chromosome haplogroups. A total of 12 haplogroups were obtained in 1214 male samples from all the nationalities. The genetic relationships among 26 nationalities were studied. The ethnic groups were compared according to their different ancient lineages. The ancient lineages had their own characteristics in the distribution of Y chromosome haplogroups. Our results showed that Yunnan Province has great genetic diversity in its people. The ethnic groups differ from each other in the number of haplogroups and haplogroup frequencies. The genetic evidence was in agreement with the study of linguistic and historical records. PMID- 15788365 TI - Metabolism of benzamidoxime (N-hydroxyamidine) in human hepatocytes and role of UDP-glucuronosyltransferases. AB - N-Hydroxyamidines (amidoximes) can act as pro-drugs of amidines (e.g. ximelagatran, a novel direct thrombin inhibitor). This known pro-drug principle is based on the N-reduction of an oral bioavailable amidoxime to its active form. Previous study of the metabolism of the model substrate benzamidoxime by pig hepatocytes demonstrated the formation of benzamidoxime-O-glucuronide in addition to the well-established N-reduction. The objective of the present work was to investigate the glucuronidation of benzamidoxime by using cultivated cryopreserved human hepatocytes. Furthermore, the involvement of human UDP glucuronosyltransferases (UGTs) was examined by incubating benzamidoxime in the presence of eight human hepatic recombinant UGT enzymes. Metabolites were analysed by liquid chromatography/mass spectrometry using electrospray ionization and compared with authentic synthetic compounds. For the first time, the O glucuronidation of benzamidoxime was demonstrated in cultures of human hepatocytes. UGT1A9 is the most efficient enzyme conjugating benzamidoxime, whereas the conversion activities of UGT1A1 and UGT1A3 were 60-fold lower. Human hepatocytes form two non-mutagenic compounds: benzamidine, as the predominating metabolite, and benzamidoxime-O-glucuronide to a lesser extent. N-oxidation of benzamidine was not detected. PMID- 15788364 TI - Comparison of clearance predictions using primary cultures and suspensions of human hepatocytes. AB - Various incubation conditions of human hepatocytes were compared for their accuracy in predicting the in vivo hepatic clearance (CL(H)) of model compounds. The test compounds were the highly cleared, low protein bound naloxone (in vivo CL(H) = 25 ml min(-1) kg(-1); free fraction = 0.6), the medium clearance, highly protein bound midazolam (CL(H) = 12 ml min(-1) kg(-1); free fraction = 0.04) and the low clearance, highly protein bound bosentan (CL(H) = 3.9 ml min(-1) kg(-1); free fraction = 0.02). Each compound was tested in three 'hepatocyte systems', using resections from three donors, in the presence and absence of human serum. Those hepatocyte systems were: conventional primary cultures, freshly isolated suspensions and cryopreserved suspended hepatocytes. Except for a twofold overestimated CL(H) for bosentan from conventional primary cultures, and despite variable cryopreservation recoveries, similar predictions of CL(H) were recorded with all hepatocyte systems. Moreover, the CL(H) values obtained with cryopreserved suspended hepatocytes were similar to those obtained with freshly isolated suspensions. For midazolam and bosentan, the predicted in vivo CL(H) was markedly higher in the presence of serum, whereas serum had little influence on the scaled-up CL(H) of naloxone. In vivo, CL(H) was properly approached for naloxone and bosentan (particularly from experiments in the presence of serum), but it was strongly underestimated for midazolam (particularly in the absence of serum). Additional compounds need to be investigated to confirm the above findings as well as to assess why the clearances of some highly protein-bound compounds are still considerably underestimated. PMID- 15788362 TI - The Provincial High Court of Barcelona. PMID- 15788366 TI - Stereoselective inhibition of cytochrome P450 forms by lansoprazole and omeprazole in vitro. AB - The stereoselectivity of the inhibitory interaction potential of lansoprazole and omeprazole isomers on six human cytochrome P450 forms was evaluated using human liver microsomes. Lansoprazole enantiomers showed stereoselective inhibition of CYP2C9-catalysed tolbutamide 4-methylhydroxylation, CYP2C19-catalysed S mephenytoin 4'-hydroxylation, CYP2D6-catalysed dextromethorphan O-demethylation, CYP2E1-catalysed chlorzoxazone 6-hydroxylation and CYP3A4-catalysed midazolam 1 hydroxylation, whereas omeprazole only inhibited CYP2C19 stereoselectively. Of the P450 forms tested, CYP2C19-catalysed S-mephenytoin 4'-hydroxylation was extensively inhibited by both the lansoprazole and omeprazole enantiomers in a competitive and stereoselective manner; the S-enantiomers of both drugs inhibited the hydroxylation more than the R-enantiomers. The estimated K(i) values determined for CYP2C19-catalysed S-mephenytoin 4'-hydroxylation were 0.6, 6.1, 3.4 and 5.7 microM for S-lansoprazole, R-lansoprazole, S-omeprazole and R omeprazole, respectively. The results indicate that although both lansoprazole and omeprazole are strong inhibitors of CYP2C19, the inhibition of CYP2C19 by lansoprazole is highly stereoselective, whereas the inhibition by omeprazole is less stereoselective. In addition, S-lansoprazole, the most potent CYP2C19 inhibitor, is not a good CYP2C19-selective inhibitor owing to its inhibition of other P450 forms. PMID- 15788367 TI - Cytochrome P450-dependent metabolism of gefitinib. AB - The in vitro metabolism of [(14)C]-gefitinib (1-3 microM) was investigated using human liver microsomes and a range of expressed human cytochrome P450 enzymes, with particular focus on the formation of O-desmethyl-gefitinib (M523595), the major metabolite observed in human plasma. High-performance liquid chromatography with ultraviolet light, radiochemical and mass spectral analysis, together with the availability of authentic standards, enabled quantification and structural identification of metabolites. On incubation with pooled human liver microsomes, [(14)C]-gefitinib underwent rapid and extensive metabolism to a number of metabolites, although M523595 was only a minor microsomal product. Formation of most metabolites was markedly decreased by ketoconazole, but M523595 production was inhibited only by quinidine. Gefitinib was metabolized extensively by expressed CYP3A4, producing a similar range of metabolites to liver microsomes, but M523595 was not formed. CYP1A2, 2C9 and 2C19 produced no measurable metabolism of gefitinib, while CYP3A5 produced a range of metabolites similar to CYP3A4, but to a much lower degree. In contrast, CYP2D6 catalysed rapid and extensive metabolism of gefitinib to M523595. While formation of M523595 was CYP2D6 mediated, the overall metabolism of gefitinib was dependent primarily on CYP3A4, and this was not obviously diminished in liver microsomes from CYP2D6 poor metabolizers. PMID- 15788368 TI - AhR and PPARalpha: antagonistic effects on CYP2B and CYP3A, and additive inhibitory effects on CYP2C11. AB - The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates a spectrum of toxic and biological effects of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. The peroxisome proliferator-activated receptor alpha (PPARalpha) is a member of the nuclear receptor super-family of ligand-activated transcription factors and it functions as an obligate heterodimer with retinoid X-receptor alpha RXRalpha. The aim was to investigate whether the negative cross-talk recently proposed by the present authors between AhR and PPARalpha on CYP4A and CYP1A has any impact on other cytochrome P450 enzymes. Treatment of male Wistar rats with a PPARalpha ligand clofibric acid (CA) induced CYP2B1/2 and CYP3A proteins, activities, and the mRNA expression of CYP2B1, CYP2B2, CYP3A1 and CYP3A2, and suppressed CYP2C11 protein, activities and mRNA expression. AhR ligand Sudan III (S.III) treatment decreased basal and CA-induced CYP2B, CYP3A and CYP2C11 protein, activities and mRNA expression. To the best of the authors' knowledge, this is the first study showing the presence of mutual effects of AhR and PPARalpha on CYP2B and CYP3A and an additive inhibitory effect on CYP2C11 in the livers of male rats. PMID- 15788369 TI - Ethnic differences between Japanese and Caucasians in the expression levels of mRNAs for CYP3A4, CYP3A5 and CYP3A7: lack of co-regulation of the expression of CYP3A in Japanese livers. AB - Using a newly developed real-time reverse transcriptase-polymerase chain reaction method, mRNAs were quantitated for CYP3A4, CYP3A5 and CYP3A7 in adult livers from 24 Japanese and 24 Caucasian subjects to elucidate the potential ethnic differences in the expression levels of human cytochrome P450 (CYP) 3As. The expression level of CYP3A4 mRNA in Japanese livers (n = 24) was approximately three times higher than that in Caucasian livers (n = 24, p < 0.001). The mean level of CYP3A5 mRNA was approximately twice higher in Japanese (n = 9) than in Caucasians (n = 5) heterozygous for the CYP3A5 *1 allele (p = 0.057). The CYP3A7 mRNA level was twice higher in Japanese (n = 24) than in Caucasians (n = 22) carrying the CYP3A7 *1A/ *1A genotype (p = 0.042). The level of CYP3A4 mRNA did not correlate with those of CYP3A5 (r = 0.044, n = 24) or CYP3A7 (r = 0.21, n = 24) mRNAs in Japanese livers in contrast to co-regulatory expression of CYP3A4, CYP3A5 and CYP3A7 in Caucasian livers. The results indicate that there are ethnic differences in the expression levels of adult liver CYP3A mRNAs between Japanese and Caucasians, and that the mechanism(s) regulating the hepatic CYP3A expression may be different between these ethnic groups. PMID- 15788370 TI - Species differences in the tissue distribution of catechol and methylsulphonyl metabolites of 2,4,5,2',5'-penta- and 2,3,4,2',3',6'-hexachlorobiphenyls in rats, mice, hamsters and guinea pigs. AB - Polychlorinated biphenyls (PCBs) are metabolized to phenolic or methylsulphonyl PCBs (MeSO(2)-CBs) in animal species. The study determined the species differences in the tissue distribution of persistent PCB metabolites in rats, mice, hamsters and guinea pigs 4 days after exposure to 2,4,5,2('),5(') pentachlorobiphenyl (CB101) or 2,3,4,2('),3('),6(')-hexachlorobiphenyl (CB132). For CB101 metabolism, the hydroxylation in rats, mice and hamsters occurred primarily at the 3(')-position in the 2('),5(')-dichlorinated phenyl ring, whereas the hydroxylation in guinea pigs occurred preferentially at the 3 position. Metabolite profiles in tissues of hamsters were dominated by 3('),4(') catechol-CB101, whereas metabolite profiles in rats and mice were dominated by 3(')- or 4(')-MeSO(2)-CBs. For CB132 metabolism, rats and mice produced 4(')- and 5(')-MeSO(2)-CBs at similar concentration ratios, whereas guinea pigs produced MeSO(2)-CBs at higher levels and selectively retained 5(')-MeSO(2)-CB in liver. In contrast, hamsters preferentially produced 4('),5(')-catechol-CB132 that was retained in serum. Consequently, hamsters produced catechols, whereas guinea pigs produced meta-substituted MeSO(2)-CBs, preferentially from CB132. These findings indicate that PCBs with 2,3,6-chlorine substitution are preferred substrates for the formation of catechols or MeSO(2)-CBs and the differences in metabolite profiles are related to species-dependent metabolic capacities. PMID- 15788372 TI - Isolation, identification and excretion profile of the principal urinary metabolite of the recently banned designer drug 1-(3 trifluoromethylphenyl)piperazine (TFMPP) in rats. AB - The metabolism of 1-(3-trifluoromethylphenyl)piperazine (TFMPP), a recently banned designer drug, in rats was studied by analysing its urinary metabolites. p Hydroxy-TFMPP (p-OH-TFMPP) was isolated and identified as the main metabolite by using nuclear magnetic resonance spectroscopy, gas chromatography-mass spectrometry and high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI MS). The time-course excretion profiles of TFMPP and p OH-TFMPP in rats were investigated following a single intraperitoneal dosing of 5 mg kg(-1) TFMPP by using an optimized analytical procedure that combined solid phase extraction and LC-ESI MS techniques. The cumulative amount of p-OH-TFMPP excreted within the first 48 h reached approximately 64% of the dose, of which 70% was the glucuronide conjugated form. The cumulative amount of parent TFMPP excreted was less than 0.7% of the dose. The results suggest that p-OH-TFMPP would be the most relevant metabolite to be detected for TFMPP exposure in the forensic and clinical analysis of human urine. PMID- 15788371 TI - Identification of glutathione conjugates and mercapturic acids of 1,2 dibromopropane in female BALB/c mice by liquid chromatography-electrospray ionization tandem mass spectrometry. AB - Based on recent results that 1,2-dibromopropane (1,2-DBP) causes hepatotoxicity and immunotoxicity in female BALB/c mice as well as a reduction of hepatic glutathione levels, the possible formation of glutathione conjugates and mercapturic acids of 1,2-DBP was investigated in vivo in the present studies. The following four metabolites were identified in the liver at 12 h after treatment with 1,2-DBP, by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS): M1, 2-hydroxypropylglutathione; M2, 2 oxopropylglutathione; M3, N-acetyl-S-(2-hydroxypropyl)-L-cysteine; and M4, N acetyl-S-(2-oxopropyl)-L-cysteine. Ions of individual conjugates were observed at m/z 366, 364, 222 and 220, respectively. Characteristic product ions at m/z 237, 217, 204 and 202 for the identification of M1, M2, M3 and M4 were observed, respectively. In the sera isolated from the same animals, only mercapturic acids (M3 and M4) were observed by LC-ESI/MS. When female BALB/c mice were treated orally with 1,2-DBP at doses of 150, 300 and 600 mg kg(-1) once for 12 h, the production of glutathione conjugates and mercapturic acids in liver was apparently dose dependent, as were the concentrations of them in sera. When the production of metabolites from 1,2-DBP was investigated in liver following oral treatment with 600 mg kg(-1) 1,2-DBP for 6, 12, 24 and 48 h, metabolite concentrations were greatest at the first time point (6 h). The results explain the authors' previous studies that oral treatment with 1,2-DBP reduces the hepatic content of glutathione. PMID- 15788373 TI - Respiratory responses to exposures with fine particulates and nitrogen dioxide in the elderly with and without COPD. AB - Elderly people, with and without chronic obstructive pulmonary disease (COPD), may be susceptible to particulate matter (PM) air pollution. However, the respiratory impacts of inhaled PM combined with copollutant(s) in controlled exposure studies are unclear and warrant investigation since exposures to PMgas mixtures constitute realistic scenarios. Thus, we exposed 6 healthy subjects and 18 volunteers with COPD (mean age 71 yr) on separate days to (a) filtered air (FA); (b) 0.4 ppm NO2; (c) concentrated ambient particles (CAP), predominantly in the fine (PM2.5) size range, at concentrations near 200 microg/m3; and (d) CAP and NO2 together. Each 2-h exposure included exercise for 15 min every half hour. Most respiratory responses, including symptoms, spirometry, and total and differential counts of induced sputum cells, showed no statistically significant responses attributable to separate or combined effects of CAP and NO2. However, maximal mid-expiratory flow and arterial O2 saturation (measured by pulse oximetry) showed small but statistically significant decrements associated with CAP, greater in healthy than COPD subjects. CAP exposure was also associated with decreased percentages of columnar epithelial cells in sputum. The results suggest that the respiratory effect of the PMNO2 mixture may be primarily PM driven since coexposure to NO2 did not significantly enhance the responses. In conclusion, older adults exposed to urban fine particles may experience acute small-airways dysfunction with impaired gas exchange. Healthy subjects appear more susceptible, suggesting that the respiratory effect may be related to efficient penetration and deposition of inhaled toxic particles in distal small airways. More clinical investigation of the elderly population is warranted. PMID- 15788374 TI - Dose dependency of adjuvant activity of particulate matter from five European sites in three seasons in an ovalbumin-mouse model. AB - Various particulate matter (PM) samples were tested for their adjuvant potency in an animal model of allergy (ovalbumin) in the European Union study entitled Respiratory Allergy and Inflammation Due to Ambient Particles. Coarse and fine ambient particles were collected during spring, summer, and winter in Rome, Oslo, Lodz, Amsterdam, and De Zilk. De Zilk, at the Dutch seaside, has mainly westerly winds and served as a negative pollution control. EHC-93 (Ottawa dust) was used as a positive control. We studied the adjuvant potency of the particle antibody responses to ovalbumin and histopathological changes in the lung. After a sensitization phase by coexposure to EHC-93 and ovalbumin, the antibody response to ovalbumin and inflammatory responses in the lung were huge. There was more adjuvant activity in reaction to 9-mg/ml samples than to 3-mg/ml samples. A best fit analysis of these samples shows that the ambient coarse and fine particles at these sites, in combination with allergens, have severe to mild adjuvant activity in the order Lodz, Rome, Oslo, and Amsterdam. A high dose of the fine fraction was more potent than a high dose of the coarse fraction, except at De Zilk, where the reverse was true. Spring and winter PM was more potent than summer PM. Depending on the site, either a water-soluble or a water-insoluble fraction was responsible for the adjuvant activity. A concentration of 3 mg/ml is effective for screening high-activity samples, as is a concentration of 9 mg/ml for screening low-activity samples in the ovalbumin-mouse model. PMID- 15788375 TI - Seasonal allergic rhinitic and normal subjects respond differentially to nasal provocation with acetic acid vapor. AB - Individuals with seasonal allergic rhinitis (SAR) show a more marked nasal obstructive response (increases in nasal airways resistance or NAR) after provocation with chlorine gas (Cl2) than do nonrhinitic (NR) controls. We were interested in learning whether similar differential responsiveness was apparent after provocation with acetic acid vapor. Sixteen nonsmoking, nonasthmatic subjects, aged 21-63 yr, equally divided by gender and nasal allergy status, were enrolled in a single-blinded crossover study involving exposure to acetic acid (AA) vapor (15 ppm) or air for 15 min on separate days 1 wk apart. NAR was measured in triplicate before, immediately post-, and 15 min postexposure, was normalized to baseline on a given exposure day, and was expressed as Net [NAR/baseline] after acetic acid versus control (air) exposure. After log transformation to achieve normality, the mean loge of Net [NAR/baseline] was 0.22 for SAR subjects and -0.11 for NR subjects immediately postexposure (p<.05); the corresponding values were 0.24 and -0.08, respectively, at 15 min postexposure (p<.05). Inhalation of acetic acid at the (NIOSH-recommended) short-term exposure limit of 15 ppm for 15 min produces differential nasal airflow obstruction among SAR versus NR subjects, with the former showing greater physiologic reactivity to this stimulus. This differential responsiveness is consistent with our previous findings with Cl2, indicating that there may be a generalized susceptibility factor associated with allergic rhinitis. The response occurs with slight subjective nasal irritation. PMID- 15788376 TI - Effect of long-term inhalation of toner on extracellular matrix in the lungs of rats in vivo. AB - We assessed the effects of long-term inhalation of toner on the pathological changes and gene expression with the synthesis and degradation of collagenous extracellular matrix in a rat model. Female Wistar rats (10 wk old) were divided evenly into a high concentration exposure group (H: 15.2 mg/m3), a low concentration exposure group (L: 5.5 mg/m3), and a control group. The mass median aerodynamic diameter of toner was 4.5 microm. The rats were sacrificed at the termination of a 1-yr or 2-yr inhalation period. Pathological examination was performed from the left lung, and transcriptional levels of mRNA extracted from the right lung were assessed by semiquantitative reverse-transcription polymerase chain polymerase (RT-PCR). The pathological findings showed mild pulmonary fibrosis in 20% (L, 1 yr), 40% (H, 1 yr), 56% (L, 2 yr) and 62% (H, 2 yr), while lung cancer was not observed in any of the exposed groups. In the 1-yr high concentration group, gene expression of matrix metalloproteinase-2 (MMP-2) and type I collagen mRNA in the rat lungs increased, while tissue inhibitors of metalloproteinase-2 (TIMP-2) decreased. The 2-yr high-concentration group increased in message level of type I collagen and TIMP-2 but not that of MMP-2. These data suggested that results of gene expression of MMP, TIMP, and collagen in the 2-yr exposure may lead to accumulation of collagen compared to the 1-yr exposure, and that the imbalance of the expression of MMPs, TIMPs, and extracellular matrix might be associated with pulmonary fibrosis induced by toner. PMID- 15788377 TI - Proinflammatory and anti-inflammatory cytokine balance in gasoline exhaust induced pulmonary injury in mice. AB - Proinflammatory and anti-inflammatory cytokine balance and associated changes in pulmonary bronchoalveolar lavage fluid (BALF) of unleaded gasoline exhaust (GE) exposed mice were investigated. Animals were exposed to GE (1 L/min of GE mixed with 14 L/min of compressed air) using a flow-past, nose-only, dynamic inhalation exposure chamber for different durations (7, 14, and 21 days). The particulate content of the GE was found to be 0.635, +/-0.10 mg PM/m3. Elevated levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were observed in BALF of GE-exposed mice, but interleukin 1beta(IL-1beta) and the anti inflammatory cytokine interleukin-10 (IL-10) remained unaffected. GE induced higher activities of alkaline phosphatase (ALP), gamma-glutamyl transferase (gammaGT), and lactate dehydrogenase (LDH) in the BALF, indicating Type II alveolar epithelial cell injury, Clara-cell injury, and general toxicity, respectively. Total protein in the BALF increased after 14 and 21 days of exposure, indicating enhanced alveolar-capillary permeability. However, the difference in the mean was found statistically insignificant in comparison to the compressed air control. Total cell count in the BALF of GE-exposed mice ranged between 0.898 and 0.813x10(6) cells/ml, whereas the compressed air control showed 0.65x10(6) cells/mL. The histopathological changes in GE-exposed lung includes perivascular, and peribronchiolar cuffing of mononuclear cells, migration of polymorphonuclear cells in the alveolar septa, alveolar thickening, and mild alveolar edematous changes indicating inflammation. The shift in pro- and anti inflammatory cytokine balance and elevation of the pulmonary marker enzymes indicate toxic insult of GE. This study will help in our understanding of the mechanism of pulmonary injury by GE in the light of cytokine profiles, pulmonary marker enzymes, and lung architecture. PMID- 15788378 TI - Single-pass environmental chamber for quantifying human responses to airborne chemicals. AB - Despite increasing interest in the short-term effects of airborne environmental contaminants, experimental findings are generated at a very slow pace. This is due in part to the expense and complexity of most environmental chambers, which are needed for quantifying effects of wholebody exposures. We lessened this obstacle by designing, constructing, and testing a single-pass, 10-m3 stainless steel chamber. Compressed air is purified before being sent to an air dilution olfactometer, which supplies 1000 L (1 m3) per minute (referenced to STP) while maintaining 40% relative humidity (RH) and 22.6 degrees C. Precise control of all stimulus parameters is greatly simplified since air is not recirculated. Vapor phase odorant concentrations are achieved by varying the proportion of total airflow passing through one or more saturators, and are verified in real time by an infrared (IR) spectrometer. An adjoining 5-m3 anteroom is used for introducing known intensities of more chemically complex vapor and/or particulate stimuli into the chamber. Prior to the point that air is exhausted from the chamber, all components are made of stainless steel, Teflon, or glass. A LabView program contains feedback loops that achieve document chamber conditions and document performance. Additional instrumentation and computer systems provide for the automated collection of perceptual, respiratory, eye blink, heart rate, blood pressure, psychological state, and cognitive data. These endpoints are now being recorded, using this facility, in response to ranges of concentrations of propionic acid and environmental tobacco smoke. PMID- 15788379 TI - Henry Smyth Jr. award lecture--occupational exposure limits: do we need them? Who is responsible? How do we get them? AB - The Henry F. Smyth Jr. Award is presented to an individual who has recognized the needs of the industrial hygiene profession and has made major contributions to fulfill those needs, thereby contributing to the improvement of the public welfare. The American Academy of Industrial Hygiene (now the Academy of Industrial Hygiene) established the Henry F. Smyth Jr. Award in 1981. Henry F. Smyth Jr. was a dedicated teacher and a productive researcher whose projects enhanced the profession. This year's Award was presented to Harry Ettinger, CIH, PE, at the 2004 Professional Conference on Industrial Hygiene (PCIH) in Montreal, Canada. PMID- 15788381 TI - Prevalence of musculoskeletal disorders in union ironworkers. AB - The prevalence of musculoskeletal disorder (MSD) symptoms and doctor-diagnosed musculoskeletal disorders (DDMDs) were estimated among union construction ironworkers by a telephone-administered questionnaire. Of 1996 ironworkers eligible, 1566 were contacted and 981 were interviewed. The prevalence of self reported MSD symptoms was high for the lower back (56%), wrist/hands/fingers (40%), knees (39%), and shoulders (36%). The most common DDMDs were tendonitis (19%), ruptured disk in the back (18%), bursitis in the shoulder (15%), and carpal tunnel syndrome (12%). Generally, the prevalence of DDMDs and MSD symptoms increased with duration of employment. In age-adjusted logistic regression analyses, those who worked 25 to 35 years were more likely to have tendonitis (odds ratio [OR] 7.1, 95% confidence interval [CI] 3.116.6), shoulder bursitis (OR 13.7, 95% CI 3.160.4), knee bursitis (OR 5.1, 95% CI 1.025.1), and ruptured intervertebral back disk (OR 6.7, 95% CI 2.617.5). The effect of prior injury was also consistently high (upper extremities, OR 4.6; lower extremities OR 5.1; lower back, OR 6.0). Among workers without prior injuries, MSD symptoms were more frequent for the lower back in structural ironwork (OR 1.7, 95% CI 1.12.6), and for the upper extremity in concrete reinforcement ironwork (OR 1.9, 95% CI 1.22.9). These findings suggest that some musculoskeletal morbidity in construction ironworkers may be work related and thus preventable. PMID- 15788383 TI - Field performance of the RespiCon for size-selective sampling of industrial wood processing dust. AB - The RespiCon sampler is a multistage virtual impactor that simultaneously collects the ISO/CEN/ACGIH size fractions of inhalable, thoracic, and respirable particulate matter. The field performance of the device for measurement of industrial wood processing dust was evaluated against reference size-selective samplers: the IOM sampler (inhalable dust), the GK 2.69 cyclone (thoracic dust), and the SKC aluminum cyclone (respirable dust). Seventy-one sets of area samples were collected from 10 wood processing plants, with the samplers mounted either in the free-field or on a two-dimensional bluff body. The geometric mean (range) dust levels across all plants measured by the reference samplers were: inhalable, 1.35 mg/ m3 (0.11-11.06); thoracic, 0.31 mg/m3 (0.05-1.38); and respirable, 0.10 mg/m3 (0.02-0.54). In comparing the RespiCon with the reference samplers, there was no significant difference between sampling in the free-field versus bluff body modes. For inhalable dust, there was no significant difference between the RespiCon and the IOM sampler after applying a correction factor of 1.5 to the extrathoracic data obtained from the RespiCon. Without the correction factor, the RespiCon undersample inhalable dust by an average of 23%. For thoracic dust, the RespiCon was shown to oversample the extrathoracic dust fraction resulting in an overall error of 48%. A simple correction based on the inhalable and thoracic dust levels reported by the RespiCon is proposed. For respirable dust, there was a significant difference between the RespiCon and the SKC cyclone, but the data were equivocal due to imprecision in measurement of the low respirable dust concentrations encountered and the likelihood of bias in the reference sampler. Overall, the RespiCon sampler appears to be a suitable size-selective sampling device for industrial wood processing dust, although adjustments should be made to the inhalable and thoracic dust results. PMID- 15788382 TI - Rapid assessment of microbial hazards in metalworking fluids. AB - Industries that use metalworking fluids require a test method that can rapidly estimate the total number of bacteria. Such a test method would improve the ability to manage the metal working fluid by allowing near real-time decision making. The ability to manage the fluid more consistently and make critical decisions as they arise would reduce occupational exposures to contaminated metalworking fluid and likely result in fewer worker complaints and/or work stoppages. In this study, a filtration-based rapid adenosine triphosphate bioluminescence assay, which takes less than 10 min to perform, was evaluated as a test method for estimating total bacteria populations in metalworking fluid. This evaluation used two types of metalworking fluid (soluble and semisynthetic) that were inoculated using Pseudomonas aeruginosa and spoiled metalworking fluid. Daily parallel testing was completed on metalworking fluid samples using the rapid adenosine triphosphate assay and the standard plate count methods. Daily test results were evaluated by calculating r2 values using statistical correlation and regression procedures for each fluid type. Study results indicate the rapid adenosine triphosphate assay is strongly correlated to the standard plate count method for soluble and semisynthetic fluids, producing results of r2 = 0.74 and r2 = 0.89, respectively. PMID- 15788384 TI - The effectiveness of hearing protection among construction workers. AB - Effective hearing conservation programs in the construction industry are rare. Where programs are present, they often rely on workers' use of hearing protection devices (HPDs) rather than on exposure controls to reduce noise exposure levels. Dependence on HPDs for protection from high noise is problematic, as the protection provided by the HPD depends on both the HPD's attenuation level and the time the HPD is used. This article presents an analysis of data on noise exposure and hearing protection among construction workers drawn from several large datasets covering nine construction trades. A unique combination of 1-min dosimetry noise exposure levels and simultaneous self-reported use of HPDs was evaluated, as were occupational and nonoccupational HPD use data collected by questionnaire as part of a longitudinal study of noise exposure and hearing loss among apprentices. Direct measurements of HPD attenuation were also made on workers at their work site. The workers assessed in this study were found to use hearing protection less than one-quarter of the time that they were exposed above 85 dBA. Workers who reported always using HPDs in high noise on questionnaires were found to wear them only one-third of the time their exposures exceeded 85 dBA. Workers' self-reported use of HPDs during most noisy nonoccupational activities was also found to be low. Direct attenuation measurements found that workers were able to achieve more than 50% of the rated attenuation of their HPD on average, but that the variability in achieved attenuation was large. When the measured HPD attenuation levels and use time data were combined, the effective protection afforded by HPDs was less than 3 dB, a negligible amount given the high exposure levels associated with construction work. However, there was substantial variation in effective protection among the different trades assessed. These results demonstrate the need for better hearing conservation programs and expanded noise control efforts in the construction industry. PMID- 15788385 TI - Evaluation of the NIOSH MWF total particulate matter: thoracic particulate matter conversion factor in a machining environment. AB - Worker exposures to metalworking fluids were characterized at a plant that produced air compressors. Full-shift, side-by-side air samples (n = 147) were collected and analyzed for total particulate matter, extractable total particulate matter, thoracic particulate matter, and extractable thoracic particulate matter. The thoracic particulate matter geometric mean of 0.32 m/m (3)was below the National Institute for Occupational Safety and Health (NIOSH) recommended exposure limit (REL) of 0.4 mg/m (3). The total particulate matter geometric mean of 0.52 mg/m (3), however, was above 0.5 mg/m (3), the total particulate matter concentration offered as a surrogate REL in the NIOSHCriteria for a Recommended Standard for Occupational Exposure to Metalworking Fluids.[1]Of the 83 total particulate matter results that were at or above smash 0.5 mg/m (3), only 50 (60%) of the corresponding thoracic particulate matter results were at or above 0.4 mg/m (3). These data indicated a conversion factor of 1.65 between thoracic particulate matter and total particulate matter concentrations and 1.40 between thoracic extractable particulate matter and total extractable concentrations. These factors were significantly different from the 1.25 used to compare total particulate matter with thoracic particulate matter concentrations in the NIOSHCriteria Document[1](p < 0.01) and call into question the validity of a universal conversion factor. The authors conclude that thoracic particulate matter exposure assessment should be done directly. In terms of protecting the worker, however, the 1.25 conversion factor appeared to be conservative since each time a total particulate matter result was below 0.5 mg/m (3), its paired thoracic particulate matter measurement was below 0.4 mg/m (3). PMID- 15788386 TI - Laboratory and field validation of the GC-NPD method for the measurement of formaldehyde in the workplace. AB - Formaldehyde is classified as a suspected or probable human carcinogen by several organizations. Since conventional sampling and analytical methods for airborne formaldehyde show relatively poor sensitivity, an improved method is needed. The aim of this study was to develop a new analytical method for measuring the airborne formaldehyde concentrations in workplaces and to evaluate the performance of the method through laboratory and field tests. The method employs a sampling tube containing silica gel coated with 2, 4-dinitrophenylhydrazine (2,4-DNPH), which produces 2,4-DNPH-formaldehyde derivative with formaldehyde. Then the 2,4-DNPH-formaldehyde derivative is analyzed using gas chromatography (GC) equipped with a nitrogen-phosphorus detector (GC-NPD). In laboratory tests, the new method, referred to as the GC-NPD method, was as sensitive as the National Institute for Occupational Safety and Health (NIOSH) analytical method, which uses high-performance liquid chromatography equipped with ultraviolet detector. The total analytical precision and 95% confidence limit of the estimated total standard error for the GC-NPD method were 0.009 and +/- 12.0%, respectively, which satisfied the OSHA sampling and analytical criteria. In field tests, the overall uncertainty of the GC-NPD method was 11.2%, which satisfied the NIOSH criteria for sampling and analytical methods. The GC-NPD method with a 2,4-DNPH coated adsorbent sampler for the determination of airborne formaldehyde concentration showed good performance with acceptable accuracy and precision. PMID- 15788387 TI - Regulation of expression of the chorionic gonadotropin/luteinizing hormone receptor gene in the human myometrium: involvement of specificity protein-1 (Sp1), Sp3, Sp4, Sp-like proteins, and histone deacetylases. AB - At present there is little information on the regulatory processes by which the chorionic gonadotropin (CG)/LH receptor gene is regulated in the human myometrium during pregnancy and labor. Employing human primary myometrial cell cultures in conjunction with DNA affinity purification assays/Western analysis, DNA binding studies, CG/LH promoter luciferase reporter gene deletion constructs in transfection assays, and measurement of endogenous mRNA levels in vivo by duplex RT-PCR, we have determined the role that the major transcriptional regulatory sequences from the +1 ATG codon to -2678 bp play in modulating expression of the CG/LH receptor gene in the myometrium. We report that the distal -180 to -2678 bp region of the promoter, although capable of binding members of the Jun family via the multiple activator protein-1 sites within this region, has no significant role in regulating the expression of the CG/LH receptor gene in myometrial cells. In contrast, the two specificity protein-1 to -4 (Sp1-4) GC boxes within the +1 to -180 bp proximal promoter are central to expression of the gene in the myometrium. However, not only are Sp1/Sp3 proteins involved in this process, but Sp4 and a novel Sp-like factor(s) also have an intimate part in transcriptional regulation of the gene. It would appear that Sp1/Sp3/Sp4 and Sp-like proteins are involved in recruiting histone deacetylase complexes to the proximal promoter, preventing chromatin remodeling resulting in transcriptional repression of the gene. Our data suggest that administration of the histone deacetylase inhibitor trichostatin A to human myometrial cells in vitro and in vivo substantially removes this silencing effect on expression of the gene and may implicate the use of this and similar agents in increasing myometrial CG/LH receptor levels and subsequent maintenance of uterine relaxation during fetal maturation. PMID- 15788389 TI - Crystal structures of undecaprenyl pyrophosphate synthase in complex with magnesium, isopentenyl pyrophosphate, and farnesyl thiopyrophosphate: roles of the metal ion and conserved residues in catalysis. AB - Undecaprenyl pyrophosphate synthase (UPPs) catalyzes the consecutive condensation reactions of a farnesyl pyrophosphate (FPP) with eight isopentenyl pyrophosphates (IPP), in which new cis-double bonds are formed, to generate undecaprenyl pyrophosphate that serves as a lipid carrier for peptidoglycan synthesis of bacterial cell wall. The structures of Escherichia coli UPPs were determined previously in an orthorhombic crystal form as an apoenzyme, in complex with Mg(2+)/sulfate/Triton, and with bound FPP. In a further search of its catalytic mechanism, the wild-type UPPs and the D26A mutant are crystallized in a new trigonal unit cell with Mg(2+)/IPP/farnesyl thiopyrophosphate (an FPP analogue) bound to the active site. In the wild-type enzyme, Mg(2+) is coordinated by the pyrophosphate of farnesyl thiopyrophosphate, the carboxylate of Asp(26), and three water molecules. In the mutant enzyme, it is bound to the pyrophosphate of IPP. The [Mg(2+)] dependence of the catalytic rate by UPPs shows that the activity is maximal at [Mg(2+)] = 1 mm but drops significantly when Mg(2+) ions are in excess (50 mm). Without Mg(2+), IPP binds to UPPs only at high concentration. Mutation of Asp(26) to other charged amino acids results in significant decrease of the UPPs activity. The role of Asp(26) is probably to assist the migration of Mg(2+) from IPP to FPP and thus initiate the condensation reaction by ionization of the pyrophosphate group from FPP. Other conserved residues, including His(43), Ser(71), Asn(74), and Arg(77), may serve as general acid/base and pyrophosphate carrier. Our results here improve the understanding of the UPPs enzyme reaction significantly. PMID- 15788388 TI - Mechanism of the maturation process of SARS-CoV 3CL protease. AB - Severe acute respiratory syndrome (SARS) is an emerging infectious disease caused by a novel human coronavirus. Viral maturation requires a main protease (3CL(pro)) to cleave the virus-encoded polyproteins. We report here that the 3CL(pro) containing additional N- and/or C-terminal segments of the polyprotein sequences undergoes autoprocessing and yields the mature protease in vitro. The dimeric three-dimensional structure of the C145A mutant protease shows that the active site of one protomer binds with the C-terminal six amino acids of the protomer from another asymmetric unit, mimicking the product-bound form and suggesting a possible mechanism for maturation. The P1 pocket of the active site binds the Gln side chain specifically, and the P2 and P4 sites are clustered together to accommodate large hydrophobic side chains. The tagged C145A mutant protein served as a substrate for the wild-type protease, and the N terminus was first digested (55-fold faster) at the Gln(-1)-Ser1 site followed by the C terminal cleavage at the Gln306-Gly307 site. Analytical ultracentrifuge of the quaternary structures of the tagged and mature proteases reveals the remarkably tighter dimer formation for the mature enzyme (K(d) = 0.35 nm) than for the mutant (C145A) containing 10 extra N-terminal (K(d) = 17.2 nM) or C-terminal amino acids (K(d) = 5.6 nM). The data indicate that immature 3CL(pro) can form dimer enabling it to undergo autoprocessing to yield the mature enzyme, which further serves as a seed for facilitated maturation. Taken together, this study provides insights into the maturation process of the SARS 3CL(pro) from the polyprotein and design of new structure-based inhibitors. PMID- 15788391 TI - Interactions of mitochondria-targeted and untargeted ubiquinones with the mitochondrial respiratory chain and reactive oxygen species. Implications for the use of exogenous ubiquinones as therapies and experimental tools. AB - Antioxidants, such as ubiquinones, are widely used in mitochondrial studies as both potential therapies and useful research tools. However, the effects of exogenous ubiquinones can be difficult to interpret because they can also be pro oxidants or electron carriers that facilitate respiration. Recently we developed a mitochondria-targeted ubiquinone (MitoQ10) that accumulates within mitochondria. MitoQ10 has been used to prevent mitochondrial oxidative damage and to infer the involvement of mitochondrial reactive oxygen species in signaling pathways. However, uncertainties remain about the mitochondrial reduction of MitoQ10, its oxidation by the respiratory chain, and its pro-oxidant potential. Therefore, we compared MitoQ analogs of varying alkyl chain lengths (MitoQn, n = 3-15) with untargeted exogenous ubiquinones. We found that MitoQ10 could not restore respiration in ubiquinone-deficient mitochondria because oxidation of MitoQ analogs by complex III was minimal. Complex II and glycerol 3-phosphate dehydrogenase reduced MitoQ analogs, and the rate depended on chain length. Because of its rapid reduction and negligible oxidation, MitoQ10 is a more effective antioxidant against lipid peroxidation, peroxynitrite and superoxide. Paradoxically, exogenous ubiquinols also autoxidize to generate superoxide, but this requires their deprotonation in the aqueous phase. Consequently, in the presence of phospholipid bilayers, the rate of autoxidation is proportional to ubiquinol hydrophilicity. Superoxide production by MitoQ10 was insufficient to damage aconitase but did lead to hydrogen peroxide production and nitric oxide consumption, both of which may affect cell signaling pathways. Our results comprehensively describe the interaction of exogenous ubiquinones with mitochondria and have implications for their rational design and use as therapies and as research tools to probe mitochondrial function. PMID- 15788390 TI - Solution NMR structure of the 48-kDa IIAMannose-HPr complex of the Escherichia coli mannose phosphotransferase system. AB - The solution structure of the 48-kDa IIA(Man)-HPr complex of the mannose branch of the Escherichia coli phosphotransferase system has been solved by NMR using conjoined rigid body/torsion angle-simulated annealing on the basis of intermolecular nuclear Overhauser enhancement data and residual dipolar couplings. IIA(Man) is dimeric and has two symmetrically related binding sites per dimer for HPr. A convex surface on HPr, formed primarily by helices 1 and 2, interacts with a deep groove at the interface of the two subunits of IIA(Man). The interaction surface on IIA(Man) is predominantly helical, comprising helix 3 from the subunit that bears the active site His-10 and helices 1, 4, and 5 from the other subunit. The total buried accessible surface area at the protein protein interface is 1450 A(2). The binding sites on the two proteins are complementary in terms of shape and distribution of hydrophobic, hydrophilic, and charged residues. The active site histidines, His-10 of IIA(Man) and His-15 (italics indicate HPr residues) of HPr, are in close proximity. An associative transition state involving a pentacoordinate phosphoryl group with trigonal bipyramidal geometry bonded to the N-epsilon2 atom of His-10 and the N-delta1 atom of His-15 can be readily formed with negligible displacement in the backbone coordinates of the residues immediately adjacent to the active site histidines. Comparing the structures of complexes of HPr with three other structurally unrelated phosphotransferase system proteins, enzymes I, IIA(glucose), and IIA(mannitol), reveals a number of common features that provide a molecular basis for understanding how HPr specifically recognizes a wide range of diverse proteins. PMID- 15788392 TI - Trapping of an assembly intermediate of photosynthetic LH1 antenna beyond B820 subunit. Significance for the assembly of photosynthetic LH1 antenna. AB - Most photosynthetic LH1 antennae undergo dissociation into B820 subunits, suggesting their universal character as structural modules. However, dissociation into subunits seems to occur reversibly only in the absence of carotenoids and the subunits were never found to bind carotenoids. The interactions of carotenoids with B820 have been studied in a newly developed reconstitution assay of the LH1 antenna from Rhodospirillum rubrum (Fiedor, L., Akahane, J., and Koyama, Y. (2004) Biochemistry 43, 16487-16496). These model studies show that B820 subunits strongly interact with carotenoids and spontaneously form stable LH1-like complexes with substoichiometric carotenoid content. This is the first experimental evidence that B820 may occur as a short-lived intermediate in the assembly of LH1 in vivo. The resulting complex of B820 subunits with carotenoid, termed iB873, is homogeneous, according to ion exchange chromatography and reproducible pigment composition. The iB873-bound carotenoid is as efficient in energy transfer to bacteriochlorophyll as the one in native antenna. To our knowledge, iB873 is the first complex binding functional carotenoid, with the spectral and biochemical properties intermediate between that of B820 and the fully assembled LH1. PMID- 15788393 TI - Toll-like receptor signaling stimulates cell cycle entry and progression in fibroblasts. AB - Toll-like receptors (TLRs) are proteins involved in recognition of foreign pathogen-associated molecular patterns and activation of processes leading to innate immune recognition. We show that stimulation of fibroblasts with a TLR5 ligand, flagellin, can induce proliferation of serum-starved cells or prevent cell cycle exit upon serum withdrawal independently of autologous growth factor secretion. Other TLR ligands, such as poly(I:C) and lipopolysaccharide, can have a similar effect only if the action of type I interferons is blocked. Flagellin stimulation can prevent cell cycle arrest induced by overexpression of exogenous cyclin-dependent kinase inhibitor p27. Stimulation of TLR5 and overexpression of MyD88, but not TRIF, TIRAP, or TRAM, result in p27 degradation, which can be suppressed by dominant negative Akt and mutation of the p27 C-terminal Thr(187) site. These data provide evidence for a nonimmune and cell autonomous role of TLR signaling, whereby TLR stimulation provides a positive signal for cell division. PMID- 15788394 TI - Target of rapamycin-dependent activation of S6 kinase is a central step in the transduction of nutritional signals during egg development in a mosquito. AB - Female mosquitoes are effective disease vectors, because they take blood from vertebrate hosts to obtain nutrients for egg development. Amino acid signaling via the target of rapamycin (TOR) pathway has been identified as a key requirement for the activation of egg development after a blood meal. We report the characterization of the TOR kinase and one of its major downstream targets, S6 kinase, of the yellow fever mosquito Aedes aegypti during egg development in adult females. Both TOR and S6K mRNA are expressed at high levels in the ovaries and in lower levels in fat body and other tissues. After a blood meal, the subcellular localization of TOR shifts from the cytoplasm to the plasma membrane of fat body cells. By detecting phosphothreonine 388 of mosquito S6 kinase, we show that TOR activity strongly increases in fat body and ovaries after a blood meal in vivo. Furthermore, phosphorylation of S6 kinase increases in in vitro cultured fat bodies after stimulation with amino acids. This increase is sensitive to the TOR inhibitor rapamycin in a concentration-dependent manner but not to the phosphatidylinositol 3-kinase/phosphatidylinositol 3-kinase-related kinase inhibitor LY294002, the MAPK inhibitor PD98059, or the translational inhibitor cycloheximide. RNA interference-mediated reduction of S6 kinase strongly inhibits the amino acid-induced up-regulation of the major yolk protein vitellogenin in vitro and effectively disrupts egg development after a blood meal in vivo. Our data show that TOR-dependent activation of S6 kinase is a central step in the transduction of nutritional information during egg development in mosquitoes. PMID- 15788395 TI - Pigment-pigment interactions in Lhca4 antenna complex of higher plants photosystem I. AB - The red-most fluorescence emission of photosystem I (733 nm at 4 K) is associated with the Lhca4 subunit of the antenna complex. It has been proposed that this unique spectral feature originates from the low energy absorption band of an excitonic interaction involving chlorophyll A5 and a second chlorophyll a molecule, probably B5 (Morosinotto, T., Breton, J., Bassi, R., and Croce, R. (2003) J. Biol. Chem. 278, 49223-49229). Because of the short distances between chromophores in Lhc proteins, the possibility that other pigments are involved in the red-shifted spectral forms could not be ruled out. In this study, we have analyzed the pigment-pigment interactions between nearest neighboring chromophores in Lhca4. This was done by deleting individual chlorophyll binding sites by mutagenesis, and analyzing the changes in the spectroscopic properties of recombinant proteins refolded in vitro. The red-shifted (733 nm) fluorescence peak, the major target of this analysis, was lost upon mutations affecting sites A4, A5, and B5 and was modified by mutating site B6. In agreement with the shorter distance between chlorophylls A5 and B5 (7.9 A) versus A4 and A5 (12.2 A) in Lhca4 (Ben-Shem, A., Frolow, F., and Nelson, N. (2003) Nature 426, 630-635), we conclude that the low energy spectral form originates from an interaction involving pigments in sites A5 and B5. Mutation at site B6, although inducing a 15-nm blue-shift of the emission peak, maintains the red-shifted emission. This implies that chromophores responsible for the interaction are conserved and suggests a modification in the pigment organization. Besides the A5-B5 pair, evidence for additional pigment-pigment interactions between chlorophylls in sites B3-A3 and B6-A6 was obtained. However, these features do not affect the red most spectral form responsible for the 733-nm fluorescence emission band. PMID- 15788396 TI - The critical role of Exo84p in the organization and polarized localization of the exocyst complex. AB - The exocyst complex plays an essential role in tethering secretory vesicles to specific domains of the plasma membrane for exocytosis. However, how the exocyst complex is assembled and targeted to sites of secretion is unclear. Here, we have investigated the role of the exocyst component Exo84p in these processes. We have generated an array of temperature-sensitive yeast exo84 mutants. Electron microscopy and cargo protein traffic analyses of these mutants indicated that Exo84p is specifically involved in the post-Golgi stage of secretion. Using various yeast mutants, we systematically studied the localization of Exo84p and other exocyst proteins by fluorescence microscopy. We found that pre-Golgi traffic and polarized actin organization are required for Exo84p localization. However, none of the exocyst proteins controls Exo84p polarization. In addition, Sec3p is not responsible for the polarization of Exo84p or any other exocyst component to the daughter cell. On the other hand, several exocyst members, including Sec10p, Sec15p, and Exo70p, clearly require Exo84p for their polarization. Biochemical analyses of the exocyst composition indicated that the assembly of Sec10p, Sec15p, and Exo70p with the rest of the complex requires Exo84p. We propose that there are at least two distinct regulatory mechanisms for exocyst polarization, one for Sec3p and one for the other members, including Exo84p. Exo84p plays a critical role in both the assembly of the exocyst and its targeting to sites of secretion. PMID- 15788397 TI - Phosphorylation of serine 147 of tis21/BTG2/pc3 by p-Erk1/2 induces Pin-1 binding in cytoplasm and cell death. AB - Treatment of U937 cells with epidermal growth factor (EGF) induces phosphorylation of tis21 and subsequent interaction of tis21 with Pin-1, resulting in the increased cell death with mitochondrial depolarization. Ser147 and Ser149 residues of tis21 were strongly phosphorylated by p-Erk1/2 and p p38(MAPK), respectively, but not by JNK. To investigate the significance of phosphorylation of the Ser147 residue, Pin-1, one of the mitotic regulators that binds to the Ser(P)/Thr(P)-Pro region, was employed. Wild type tis21 phosphorylated by p-Erk1/2 clearly increased its binding to Pin-1, but not the P148A mutant, indicating that Pin-1 was bound to the Ser(P)147-Pro148 region of tis21. Transfection of tis21 significantly enhanced EGF-induced Pin-1 diffusion to cytoplasm, compared with that in the vector-transfected cells. Knockdown of tis21 expression by using shRNAi significantly inhibited EGF-induced Pin-1 diffusion, and analysis by flow cytometry after JC-1 stain and confocal microscope revealed that EGF aggravated tis21-induced mitochondrial depolarization and cell death. Furthermore, tis21 was bound to cyclin B1 and Cdc2 and inhibited its activity in vivo and in vitro. In summary, treatment of U937 cells with EGF activates Erk1/2, which in turn phosphorylates Ser147 of tis21 and induces tis21 and Pin-1 binding and mitochondrial depolarization. These data suggest, for the first time, a mechanism of how EGF can be antiproliferative in human tumor cells: binding of tis21/BTG2/pc3 to Pin-1 or cyclin B1-Cdc2 complex and induction of mitochondrial depolarization. PMID- 15788398 TI - Alternative splicing of vitamin D-24-hydroxylase: a novel mechanism for the regulation of extrarenal 1,25-dihydroxyvitamin D synthesis. AB - Synthesis of the active form of vitamin D, 1,25-dihydroxyvitamin D (1,25 (OH)(2)D), by renal epithelial cells is tightly controlled during normal calcium homeostasis. By contrast, macrophage production of 1,25-(OH)(2)D is often dysregulated with potential hypercalcemic complications. We have postulated that this is due to abnormal catabolism of 1,25-(OH)(2)D by the feedback control enzyme, vitamin D-24-hydroxylase (CYP24). Using chick HD-11 and human THP-1 myelomonocytic cell lines, we have shown that macrophage-like cells express a splice variant of the CYP24 gene (CYP24-SV), which encodes a truncated protein. Compared with the holo-CYP24 gene product in chick and human cells (508 and 513 amino acids, respectively), the truncated CYP24-SV versions consisted of 351 and 372 amino acids. These CYP24-SV proteins retained intact substrate-binding domains but lacked mitochondrial targeting sequences and were therefore catalytically inactive. In common with CYP24, expression of the CYP24 variants was induced by 1,25-(OH)(2)D but without a concomitant rise in 24-hydroxylase activity. However, overexpression of CYP24-SV in HD-11 and THP-1 cells reduced synthesis of 1,25-(OH)(2) D (40-50%), whereas antisense CYP24-SV expression increased 1,25-(OH)(2)D production by 2-7-fold. These data suggest that alternative splicing of CYP24 leads to the generation of a dominant negative acting protein that is catalytically dysfunctional. We theorize that expression of the CYP24-SV may contribute to the extracellular accumulation of 1,25(OH)(2)D in human health and disease. PMID- 15788399 TI - Recombinant porcine zona pellucida glycoproteins expressed in Sf9 cells bind to bovine sperm but not to porcine sperm. AB - The zona pellucida, which surrounds the mammalian oocyte, consists of the ZPA, ZPB, and ZPC glycoproteins and plays roles in species-selective sperm-egg interactions via its carbohydrate moieties. In the pig, this activity is conferred by tri- and tetraantennary complex type chains; in cattle, it is conferred by a chain of 5 mannose residues. In this study, porcine zona glycoproteins were expressed as secreted forms, using the baculovirus-Sf9 insect cell system. The sperm binding activities of the recombinant proteins were examined in three different assays. The assays clearly demonstrated that recombinant ZPB bound bovine sperm weakly but did not bind porcine sperm; when recombinant ZPC was also present, bovine sperm binding activity was greatly increased, but porcine sperm still was not bound. The major sugar chains of ZPB were pauci and high mannose type chains that were similar in structure to the major neutral N-linked chain of the bovine zona. In fact, the nonreducing terminal alpha-mannose residues were necessary for the sperm binding activity. These results show that the carbohydrate moieties of zona glycoproteins, but not the polypeptide moieties, play an essential role in species-selective recognition of porcine and bovine sperm. Moreover, Asn to Asp mutations at either of two of the N-glycosylation sites of ZPB, residue 203 or 220, significantly reduced the sperm binding activity of the ZPB/ZPC mixture, whereas a similar mutation at the third N-glycosylation site, Asn-333, had no effect on binding. These results suggest that the N-glycans located in the N-terminal half of the ZP domain of porcine ZPB are involved in sperm-zona binding. PMID- 15788400 TI - Nucleotide- and stoichiometry-dependent DNA supercoiling by reverse gyrase. AB - Reverse gyrase is a unique type IA topoisomerase that can introduce positive supercoils into DNA. We have investigated some of the biochemical properties of Archaeoglobus fulgidus reverse gyrase. It can mediate three distinct supercoiling reactions depending on the adenine nucleotide cofactor that is present in the reaction. Besides the ATP-driven positive supercoiling reaction, the enzyme can introduce negative supercoils with a nonhydrolyzable analog, adenylyl imidodiphosphate. In the presence of ADP the plasmid DNA is relaxed almost completely, leaving a very low level of positive supercoiling. Surprisingly, the final supercoiling extent for all three distinct reactions depends on the stoichiometry of enzyme to DNA. This dependence is not due to the difference of reaction rate, suggesting that the amount of enzyme bound to DNA is an important determinant for the final supercoiling state of the reaction product. Reverse gyrase also displays exquisite sensitivity toward temperature. Raising the reaction temperatures from 80 to 85 degrees C, both of which are within the optimal growth temperature of A. fulgidus, greatly increases enzyme activity for all the supercoiling reactions. For the reaction with AMPPNP, the product is a hypernegatively supercoiled DNA. This dramatic enhancement of the reverse gyrase activity is also correlated with the appearance of DNA in a pre-melting state at 85 degrees C, likely due to the presence of extensively unwound regions in the plasmid. The possible mechanistic insights from these findings will be presented here. PMID- 15788401 TI - Pyrrolysine and selenocysteine use dissimilar decoding strategies. AB - Selenocysteine (Sec) and pyrrolysine (Pyl) are known as the 21st and 22nd amino acids in protein. Both are encoded by codons that normally function as stop signals. Sec specification by UGA codons requires the presence of a cis-acting selenocysteine insertion sequence (SECIS) element. Similarly, it is thought that Pyl is inserted by UAG codons with the help of a putative pyrrolysine insertion sequence (PYLIS) element. Herein, we analyzed the occurrence of Pyl-utilizing organisms, Pyl-associated genes, and Pyl-containing proteins. The Pyl trait is restricted to several microbes, and only one organism has both Pyl and Sec. We found that methanogenic archaea that utilize Pyl have few genes that contain in frame UAG codons, and many of these are followed with nearby UAA or UGA codons. In addition, unambiguous UAG stop signals could not be identified. This bias was not observed in Sec-utilizing organisms and non-Pyl-utilizing archaea, as well as with other stop codons. These observations as well as analyses of the coding potential of UAG codons, overlapping genes, and release factor sequences suggest that UAG is not a typical stop signal in Pyl-utilizing archaea. On the other hand, searches for conserved Pyl-containing proteins revealed only four protein families, including methylamine methyltransferases and transposases. Only methylamine methyltransferases matched the Pyl trait and had conserved Pyl, suggesting that this amino acid is used primarily by these enzymes. These findings are best explained by a model wherein UAG codons may have ambiguous meaning and Pyl insertion can effectively compete with translation termination for UAG codons obviating the need for a specific PYLIS structure. Thus, Sec and Pyl follow dissimilar decoding and evolutionary strategies. PMID- 15788402 TI - Nuclear trapping of the forkhead transcription factor FoxO1 via Sirt-dependent deacetylation promotes expression of glucogenetic genes. AB - Activation of NAD-dependent deacetylases, or Sirtuins, prolongs life span and mimics the effects of caloric restriction in yeast. The FoxO subfamily of forkhead transcription factors has been shown to mediate some of the effects of Sirtuins. Here we have shown that Sirtuin activation or hydrogen peroxide treatment overrides the phosphorylation-dependent nuclear exclusion of FoxO1 caused by growth factors and causes nuclear translocation of FoxO1 in hepatocytes. Kinetic measurements of nuclear fluorescence recovery after photobleaching show that FoxO1 is readily diffusible within the nucleus under normal conditions but becomes restricted within a nuclear subdomain following treatment with the prototypical Sirtuin agonist resveratrol or oxidative stress. Expression of FoxO1 target genes is accordingly increased, leading to activation of gluconeogenesis and increased glucose release from hepatocytes. Selective modulation of the FoxO/Sirtuin interaction represents a promising therapeutic modality for metabolic disorders. PMID- 15788403 TI - Sequence context effects on oligo(dT) termination signal recognition by Saccharomyces cerevisiae RNA polymerase III. AB - Eukaryotic RNA polymerase (Pol) III terminates transcription at short runs of T residues in the coding DNA strand. By genomic analysis, we found that T(5) and T(4) are the shortest Pol III termination signals in yeasts and mammals, respectively, and that, at variance with yeast, oligo(dT) terminators longer than T(5) are very rare in mammals. In Saccharomyces cerevisiae, the strength of T(5) as a terminator was found to be largely influenced by both the upstream and the downstream sequence context. In particular, the CT sequence, which is naturally present downstream of T(5) in the 3'-flank of some tDNAs, was found to act as a terminator-weakening element that facilitates translocation by reducing Pol III pausing at T(5). In contrast, tDNA transcription termination was highly efficient when T(5) was followed by an A or G residue. Surprisingly, however, when a termination-proficient T(5) signal was taken out from the tDNA context and placed downstream of a fragment of the SCR1 gene, its termination activity was compromised, both in vitro and in vivo. Even the T(6) sequence, acting as a strong terminator in tRNA gene contexts, was unexpectedly weak within the SNR52 transcription unit, where it naturally occurs. The observed sequence context effects reflect intrinsic recognition properties of Pol III, because they were still observed in a simplified in vitro transcription system only consisting of purified RNA polymerase and template DNA. Our findings strengthen the notion that termination signal recognition by Pol III is influenced in a complex way by the region surrounding the T cluster and suggest that read-through transcription beyond T clusters might play a significant role in the biogenesis of class III gene products. PMID- 15788404 TI - Biochemical and molecular characterization of a novel choline-specific glycerophosphodiester phosphodiesterase belonging to the nucleotide pyrophosphatase/phosphodiesterase family. AB - Nucleotide pyrophosphatases/phosphodiesterases (NPPs) are ubiquitous membrane associated or secreted ectoenzymes that release nucleoside 5'-monophosphate from a variety of nucleotides and nucleotide derivatives. The mammalian NPP family comprises seven members, but only three of these (NPP1-3) have been studied in some detail. Previously we showed that lysophospholipase D, which hydrolyzes lysophosphatidylcholine (LPC) to produce lysophosphatidic acid, is identical to NPP2. More recently an uncharacterized novel NPP member (NPP7) was shown to have alkaline sphingomyelinase activity. These findings raised the possibility that other members of the NPP family act on phospholipids. Here we show that the sixth member of the NPP family, NPP6, is a choline-specific glycerophosphodiester phosphodiesterase. The sequence of NPP6 encodes a transmembrane protein containing an NPP domain with significant homology to NPP4, NPP5, and NPP7/alkaline sphingomyelinase. When expressed in HeLa cells, NPP6 was detected in both the cells and the cell culture medium as judged by Western blotting and by enzymatic activity. Recombinant NPP6 efficiently hydrolyzed the classical substrate for phospholipase C, p-nitrophenyl phosphorylcholine, but not the classical nucleotide phosphodiesterase substrate, p-nitrophenyl thymidine 5' monophosphate. In addition, NPP6 hydrolyzed LPC to form monoacylglycerol and phosphorylcholine but not lysophosphatidic acid, showing it has a lysophospholipase C activity. NPP6 showed a preference for LPC with short (12:0 and 14:0) or polyunsaturated (18:2 and 20:4) fatty acids. It also hydrolyzed glycerophosphorylcholine and sphingosylphosphorylcholine efficiently. In mice, NPP6 mRNA was predominantly detected in kidney with a lesser expression in brain and heart, and in human it was detected in kidney and brain. The present results suggest that NPP6 has a specific role through the hydrolysis of polyunsaturated LPC, glycerophosphorylcholine, or sphingosylphosphorylcholine in these organs. PMID- 15788406 TI - Oxysterols inhibit phosphatidylcholine synthesis via ERK docking and phosphorylation of CTP:phosphocholine cytidylyltransferase. AB - Surfactant deficiency contributes to acute lung injury and may result from the elaboration of bioactive lipids such as oxysterols. We observed that the oxysterol 22-hydroxycholesterol (22-HC) in combination with its obligate partner, 9-cis-retinoic acid (9-cis-RA), decreased surfactant phosphatidylcholine (PtdCho) synthesis by increasing phosphorylation of the regulatory enzyme CTP:phosphocholine cytidylyltransferase-alpha (CCTalpha). Phosphorylation of CCTalpha decreased its activity. 22-HC/9-cis-RA inhibition of PtdCho synthesis was blocked by PD98059 or dominant-negative ERK (p42 kinase). Overexpression of constitutively active MEK1, the kinase upstream of p42 kinase, increased CCTalpha phosphorylation. Expression of truncated CCTalpha mutants lacking proline directed sites within the C-terminal phosphorylation domain partially blocked oxysterol-mediated inhibition of PtdCho synthesis. Mutagenesis of Ser315 within CCTalpha was both required and sufficient to confer significant resistance to 22 HC/9-cis-RA inhibition of PtdCho synthesis. A novel putative ERK-docking domain N terminal to this phosphoacceptor site was mapped within the CCTalpha membrane binding domain (residues 287-300). The results are the first demonstration of a physiologically relevant phosphorylation site and docking domain within CCTalpha that serve as targets for ERKs, resulting in inhibition of surfactant synthesis. PMID- 15788405 TI - Collagen alpha1(I) gene (COL1A1) is repressed by RFX family. AB - Collagen type I is composed of three polypeptide chains transcribed from two separate genes (COL1A1 and COL1A2) with different promoters requiring coordinate regulation. Our recent publications, centering on COL1A2 regulation, demonstrate that methylation in the first exon of COL1A2 at a regulatory factor for X box (RFX) site (at -1 to +20) occurs in human cancer cells and correlates with increased RFX1 binding and decreased collagen transcription (Sengupta, P. K., Erhlich, M., and Smith, B. D. (1999) J. Biol. Chem. 274, 36649-36655; Sengupta, S., Smith, E. M., Kim, K., Murnane, M. J., and Smith, B. D. (2003) Cancer Res. 63, 1789-1797). In normal cells, RFX5 complex along with major histocompatibility class II transactivator (CIITA) is induced by interferon-gamma to occupy this site and repress collagen transcription (Xu, Y., Wang, L., Buttice, G., Sengupta, P. K., and Smith, B. D. (2004) J. Biol. Chem. 279, 41319-41332). In this paper, we demonstrate that COL1A1 has an RFX consensus binding site surrounding the transcription start site (-11 to +10) that contains three methylation sites rather than one in the COL1A2 gene RFX binding site. RFX1 interacts weakly with the unmethylated COL1A1 site, and binds with higher affinity to the methylated site. RFX1 represses the unmethylated COL1A1 less efficiently than COL1A2. COL1A1 promoter activity is sensitive to DNA methylation and the COL1A1 gene is methylated in human cancer cells with coordinately decreased collagen expression. The DNA methylation inhibitor, 5-aza-2'-deoxycytidine (aza-dC) increases collagen gene expression with time in human cancer cells. On the other hand, RFX5 interacts with both collagen type I genes with a similar binding affinity and represses both promoters equally in transient transfections. Two dominant negative forms of RFX5 activate both collagen genes coordinately. Finally, CIITA RNA interference experiments indicate that CIITA induction is required for interferon gamma-mediated repression of both collagen type I genes. PMID- 15788407 TI - Single cell analysis and temporal profiling of agonist-mediated inositol 1,4,5 trisphosphate, Ca2+, diacylglycerol, and protein kinase C signaling using fluorescent biosensors. AB - The magnitude and temporal nature of intracellular signaling cascades can now be visualized directly in single cells by the use of protein domains tagged with enhanced green fluorescent protein (eGFP). In this study, signaling downstream of G protein-coupled receptor-mediated phospholipase C (PLC) activation has been investigated in a cell line coexpressing recombinant M(3) muscarinic acetylcholine and alpha(1B) -adrenergic receptors. Confocal measurements of changes in inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3)), using the pleckstrin homology domain of PLCdelta1 tagged to eGFP (eGFP-PH(PLCdelta)), and 1,2 diacylglycerol (DAG), using the C1 domain of protein kinase Cgamma (PKCgamma) (eGFP-C1(2)-PKCgamma), demonstrated clear translocation responses to methacholine and noradrenaline. Single cell EC(50) values calculated for each agonist indicated that responses to downstream signaling targets (Ca(2+) mobilization and PKC activation) were approximately 10-fold lower compared with respective Ins(1,4,5)P(3) and DAG EC(50) values. Examining the temporal profile of second messenger responses to sub-EC(50) concentrations of noradrenaline revealed oscillatory Ins(1,4,5)P(3), DAG, and Ca(2+) responses. Oscillatory recruitments of conventional (PKCbetaII) and novel (PKCepsilon) PKC isoenzymes were also observed which were synchronous with the Ca(2+) response measured simultaneously in the same cell. However, oscillatory PKC activity (as determined by translocation of eGFP-tagged myristoylated alanine-rich C kinase substrate protein) required oscillatory DAG production. We suggest a model that uses regenerative Ca(2+) release via Ins(1,4,5)P(3) receptors to initiate oscillatory second messenger production through a positive feedback effect on PLC. By acting on various components of the PLC signaling pathway the frequency-encoded Ca(2+) response is able to maintain signal specificity at a level downstream of PKC activation. PMID- 15788408 TI - The oxidative stressor arsenite activates vascular endothelial growth factor mRNA transcription by an ATF4-dependent mechanism. AB - Aberrant retinal expression of vascular endothelial growth factor (VEGF) leading to neovascularization is a central feature of age-related macular degeneration and diabetic retinopathy, two leading causes of vision loss. Oxidative stress is suggested to occur in retinal tissue during age-related macular degeneration and diabetic retinopathy and is suspected in the mechanism of VEGF expression in these diseases. Arsenite, a thiol-reactive oxidative stressor, induces VEGF expression by a HIF-1alpha-independent mechanism. Previously, we demonstrated that homocysteine, an endoplasmic reticulum stressor, increases VEGF transcription by a mechanism dependent upon activating transcription factor ATF4. Because ATF4 is expressed in response to oxidative stress, we hypothesized that ATF4 was also responsible for increased VEGF transcription in response to arsenite. We now show that arsenite increased steady state levels of VEGF mRNA and activated transcription from a VEGF promoter construct. Arsenite induced eIF2alpha phosphorylation, resulting in increased ATF4 protein levels. Inactivation or loss of ATF4 greatly diminished the VEGF response to arsenite treatment. Overexpression of ATF4 was sufficient to activate the VEGF promoter, and arsenite cooperated with exogenous ATF4 to further activate the promoter. A complex containing ATF4 binds a DNA element at +1767 bp relative to the VEGF transcription start site, and DNA binding activity is increased by arsenite treatment. In addition, the ability of a thiol antioxidant, N-acetylcysteine, to inhibit the effect of arsenite on VEGF expression coincided with its ability to inhibit phosphorylation of eIF2alpha and ATF4 protein expression. Thus, arsenite induced up-regulation of VEGF gene transcription occurs by an ATF4-dependent mechanism. PMID- 15788409 TI - RNA CUG-binding protein 1 increases translation of 20-kDa isoform of CCAAT/enhancer-binding protein beta by interacting with the alpha and beta subunits of eukaryotic initiation translation factor 2. AB - Expression of a dominant negative 20-kDa isoform of CCAAT/enhancer-binding protein (C/EBPbeta), LIP, is increased in proliferating livers and in tumor cells. Two RNA-binding proteins, CUGBP1 and calreticulin, have been implicated in the translational regulation of C/EBPbeta. In this paper, we present evidence showing several critical steps by which liver increases translation of LIP after partial hepatectomy. At early stages after partial hepatectomy, liver activates CUGBP1 by a hyperphosphorylation. The activated CUGBP1 binds to the 5' region of C/EBPbeta mRNA and replaces calreticulin, which partially represses translation of C/EBPbeta in quiescent livers. The hyperphosphorylated CUGBP1 also interacts with the alpha and beta subunits of initiation factor eIF2. Our data demonstrate that the interaction of CUGBP1 with the eIF2alpha enhances the association of CUGBP1 with ribosomes and correlates with increased translation of LIP in the liver after partial hepatectomy. Our data support the hypothesis that CUGBP1 increases translation of LIP by the interaction with the eIF2alpha subunit. This facilitates subsequent recruitment of larger numbers of ribosomes to initiate translation of LIP. PMID- 15788410 TI - Abrogation of transforming growth factor-beta signaling by SMAD7 inhibits collagen gel contraction of human dermal fibroblasts. AB - Human fibroproliferative disorders like hypertrophic scarring of the skin are characterized by increased contractility and excess extracellular matrix synthesis. A beneficial role of transforming growth factor (TGF)-beta in wound healing was proposed; however, chronic stimulation by this cytokine leads to fibrosis. In the present report, the intracellular TGF-beta signaling in fibroblasts derived from hypertrophic scars and normal skin was examined. In an attempt to intervene in profibrogenic TGF-beta functions, ectopic expression of Smad7 or dominant negative Smads3/4 completely inhibited contractility of scar derived and normal fibroblasts after suspension in collagen gels. Both cell types displayed constitutive Smad2/3 phosphorylation and (CAGA)9-MLP-Luc activity with expression and phosphorylation of Smad3 being predominant in hypertrophic scar derived fibroblasts. Down-regulation of intrinsic signaling with various TGF-beta antagonists, e.g. soluble TGF-beta receptor, latency-associated peptide, and anti TGF-beta1 antibodies, confirms autocrine TGF-beta stimulation of both cell populations. Further, Smad7 expression inhibited alpha1 (I) collagen and alpha smooth muscle actin expression. In summary, our data indicate that autocrine TGF beta/Smad signaling is involved in contractility and matrix gene expression of fibroblasts from normal and hypertrophic scars. Smad7 inhibits these processes and may exert beneficial effects on excessive scar formation. PMID- 15788411 TI - A novel mechanism for protein delivery: granzyme B undergoes electrostatic exchange from serglycin to target cells. AB - The molecular interaction of secreted granzyme B-serglycin complexes with target cells remains undefined. Targets exposed to double-labeled granzyme B-serglycin complexes show solely the uptake of granzyme B. An in vitro model demonstrates the exchange of the granzyme from serglycin to immobilized, sulfated glycosaminoglycans. Using a combination of cell binding and internalization assays, granzyme B was found to exchange to sulfated glycosaminoglycans and, depending on the cell type, to higher affinity sites. Apoptosis induced by purified granzyme B and cytotoxic T-cells was diminished in targets with reduced cell surface glycosaminoglycan content. A mechanism of delivery is proposed entailing electrostatic transfer of granzyme B from serglycin to cell surface proteins. PMID- 15788412 TI - Crystal structure of human vacuolar protein sorting protein 29 reveals a phosphodiesterase/nuclease-like fold and two protein-protein interaction sites. AB - Vacuolar protein sorting protein 29 (Vps29p), which is involved in retrograde trafficking from prevacuolar endosomes to the trans-Golgi network, performs its biological functions by participating in the formation of a "retromer complex." In human cells, this complex comprises four conserved proteins: hVps35p, hVps29p, hVps26p, and sorting nexin 1 protein (SNX1). Here, we report the crystal structure of hVps29p at 2.1 Angstroms resolution, the first three-dimensional structure of the retromer subunits. This novel structure adopts a four-layered alpha-beta-beta-alpha sandwich fold. hVps29p contains a metal-binding site that is very similar to the active sites of some proteins of the phosphodiesterase/nuclease protein family, indicating that hVps29p may carry out chemically similar functions. Structure and sequence conservation analysis suggests that hVps29p contains two protein-protein interaction sites. One site, which potentially serves as the interface between hVps29p and hVps35p, comprises 5 conserved hydrophobic and 8 hydrophilic residues. The other site is relatively more hydrophilic and may serve as a binding interface with hVps26p, SNX1, or other target proteins. PMID- 15788413 TI - The extracellular matrix protein MAGP-2 interacts with Jagged1 and induces its shedding from the cell surface. AB - Elastic fibers are composed of the protein elastin and a network of 10-12-nm microfibrils, which are composed of several glycoproteins, including fibrillin-1, fibrillin-2, and MAGP1/2 (microfibril-associated glycoproteins-1 and -2). Although fibrillins and MAGPs covalently associate, we find that the DSL (Delta/Serrate/LAG2) protein Jagged1, an activating ligand for Notch receptor signaling, also interacts with MAGP-2 in both yeast two-hybrid and coimmunoprecipitation studies. Interaction between Jagged1 and MAGP-2 requires the epidermal growth factor-like repeats of Jagged1. MAGP-2 was found complexed with the Jagged1 extracellular domain shed from 293T cells and COS-7 cells coexpressing full-length Jagged1 and MAGP-2. MAGP-2 shedding of the Jagged1 extracellular domain was decreased by the metalloproteinase hydroxamate inhibitor BB3103 implicating proteolysis in its release. Although MAGP-2 also interacted with the other DSL ligands, Jagged2 and Delta1, they were not found associated with MAGP-2 in the conditioned media, identifying differential effects of MAGP-2 on DSL ligand shedding. The related microfibrillar protein MAGP-1 was also found to interact with DSL ligands but, unlike MAGP-2, was unable to facilitate the shedding of Jagged1. Our findings suggest that in addition to its role in microfibrils, MAGP-2 may also affect cellular differentiation through modulating the Notch signaling pathway either by binding to cell surface DSL ligands or by facilitating release and/or stabilization of a soluble extracellular form of Jagged1. PMID- 15788414 TI - Mouse large can modify complex N- and mucin O-glycans on alpha-dystroglycan to induce laminin binding. AB - The human LARGE gene encodes a protein with two putative glycosyltransferase domains and is required for the generation of functional alpha-dystroglycan (alpha-DG). Monoclonal antibodies IIH6 and VIA4-1 recognize the functional glycan epitopes of alpha-DG that are necessary for binding to laminin and other ligands. Overexpression of full-length mouse Large generated functionally glycosylated alpha-DG in Pro(-5) Chinese hamster ovary (CHO) cells, and the amount was increased by co-expression of protein:O-mannosyl N-acetylglucosaminyltransferase 1. However, functional alpha-DG represented only a small fraction of the alpha-DG synthesized by CHO cells or expressed from an alpha-DG construct. To identify features of the glycan epitopes induced by Large, the production of functionally glycosylated alpha-DG was investigated in several CHO glycosylation mutants. Mutants with defective transfer of sialic acid (Lec2), galactose (Lec8), or fucose (Lec13) to glycoconjugates, and the Lec15 mutant that cannot synthesize O mannose glycans, all produced functionally glycosylated alpha-DG upon overexpression of Large. Laminin binding and the alpha-DG glycan epitopes were enhanced in Lec2 and Lec8 cells. In Lec15 cells, functional alpha-DG was increased by co-expression of core 2 N-acetylglucosaminyltransferase 1 with Large. Treatment with N-glycanase markedly reduced functionally glycosylated alpha-DG in Lec2 and Lec8 cells. The combined data provide evidence that Large does not transfer to Gal, Fuc, or sialic acid on alpha-DG nor induce the transfer of these sugars to alpha-DG. In addition, the data suggest that human LARGE may restore functional alpha-DG to muscle cells from patients with defective synthesis of O-mannose glycans via the modification of N-glycans and/or mucin O glycans on alpha-DG. PMID- 15788415 TI - Probing the role of Asp-120(81) of metallo-beta-lactamase (IMP-1) by site directed mutagenesis, kinetic studies, and X-ray crystallography. AB - Metallo-beta-lactamase IMP-1 is a di-Zn(II) metalloenzyme that efficiently hydrolyzes beta-lactam antibiotics. Wild-type (WT) IMP-1 has a conserved Asp 120(81) in the active site, which plays an important role in catalysis. To probe the catalytic role of Asp-120(81) in IMP-1, the IMP-1 mutants, D120(81)A and D120(81)E, were prepared by site-directed mutagenesis, and various kinetics studies were conducted. The IMP-1 mutants exhibited 10(2)-10(4)-fold drops in k(cat) values compared with WT despite the fact that they contained two Zn(II) ions in the active site. To evaluate the acid-base characteristics of Asp 120(81), the pH dependence for hydrolysis was examined by stopped-flow studies. No observable pK(a) values between pH 5 and 9 were found for WT and D120(81)A. The rapid mixing of equimolar amounts of nitrocefin and all enzymes failed to result in the detection of an anion intermediate of nitrocefin at 650 nm. These results suggest that Asp-120(81) of IMP-1 is not a factor in decreasing the pK(a) for the water bridging two Zn(II) ions and is not a proton donor to the anionic intermediate. In the case of D120(81)E, the nitrocefin hydrolysis product, which shows a maximum absorption at 460 nm, was bound to D120(81)E in the protonated form. The three-dimensional structures of D120(81)A and D120(81)E were also determined at 2.0 and 3.0 A resolutions, respectively. In the case of D120(81)E, the Zn-Zn distance was increased by 0.3 A compared with WT, due to the change in the coordination mode of Glu-120(81)OE1 and the positional shift in the conserved His-263(197) at the active site. PMID- 15788416 TI - Metastasis-associated protein S100A4 induces angiogenesis through interaction with Annexin II and accelerated plasmin formation. AB - Many advanced tumors overexpress and secrete the S100A4 protein that is known to promote angiogenesis and metastasis development. The mechanisms of this effect and the endothelial receptor for S100A4 are both still unknown. Here we report that extracellular S100A4 interacts with annexin II, an endothelial plasminogen co-receptor. Co-localization and direct binding of S100A4 and annexin II were demonstrated, and the binding site was identified in the N-terminal region of annexin II. S100A4 alone or in a complex with annexin II accelerated tissue plasminogen activator-mediated plasminogen activation in solution and on the endothelial cell surface through interaction of the S100A4 C-terminal lysines with the lysine-binding domains of plasminogen. A synthetic peptide corresponding to the N terminus of annexin II prevented S100A4-induced plasmin formation in the endothelial cell culture. Local plasmin formation induced by circulating S100A4 could contribute to tumor-induced angiogenesis and metastasis formation that makes this protein an attractive target for new anti-cancer and anti-angiogenic therapies. PMID- 15788417 TI - Response of Harshaw neutron thermoluminescence dosemeters in terms of the revised ICRP/ICRU recommendations. AB - To monitor workers for external neutron radiation dose, the Y-12 National Security Complex utilises the thermoluminescence dosemeters (TLDs) manufactured by Harshaw. At Y-12, the majority of external dose to workers is due to low energy photon and/or beta particles emitted from uranium and its progeny. However, some neutron dose is expected since neutrons are produced from (alpha,n) reactions in various compounds found at the plant, including UF4 and UF6. Neutron sources, such as 252Cf, are also used throughout the complex. The Harshaw neutron dosemeter consists of two gamma-sensitive elements (7Li) and two neutron sensitive elements enriched in 6Li with various shielding/filter materials placed around each of them. In this work, the energy response of the dosemeter to neutrons has been calculated using the Monte Carlo transport code MCNP Version 4 C and, these results are compared with the measured response of the dosemeter to unmoderated and D2O-moderated 252Cf neutrons. The response of the dosemeter has also been determined in terms of the personal absorbed dose and personal dose equivalent as a function of neutron energy based on the recommendations of the ICRP Publication 60 and ICRU Report 49. The energy response of the dosemeter characteristics can be used to generate spectral conversion coefficients for routine neutron absorbed dose and dose equivalent calculations. PMID- 15788418 TI - Angular dependence of the TL reading of thin alpha-Al2O3:C dosemeters exposed to different beta spectra. AB - The angular dependence of the thermoluminescent (TL) signal of thin alpha-Al2O3:C dosemeters was investigated for a series of beta-emitting radionuclides commonly employed in nuclear medicine and characterised by different mean energies (99Tc, 177Lu, 90Sr/90Y and 90Y). Irradiations were performed in a controlled geometry, using a properly designed irradiator intended to realistically reproduce the situation of exposure of hospital personnel to beta-emitting pharmaceuticals. Under the conditions of extended source and short source to detector distance, the TL signal of thin alpha-Al2O3:C layers per unit irradiation time was observed to be independent on the angle of incidence within acceptable limits, particularly for those radionuclides with maximum energy >500 keV. This property may be easily explained by using simple physical considerations, such as the limited thickness of the dosemeters. The results confirm that these detectors are suitable for beta-ray extremity dose measurements, when the photon contribution is negligible, as in the case considered. PMID- 15788419 TI - Field comparison of two different passive radon detectors. AB - The present paper describes field performance of two different passive radon monitor devices formed, respectively, by a CR-39 track detector enclosed in a diffusion chamber and a cellulose nitrate detector (LR-115) in a heat-sealed polyethylene bag. The most important scope of these investigations was to study the performance of these detectors directly in the field. To this end, two different types of radon monitors mentioned above have been exposed simultaneously in 100 school rooms within the Italian region Friuli Venezia Giulia. Finally, the accuracy of their responses has been evaluated by exposing them under extreme humidity conditions and/or together with other radon measurement instruments. PMID- 15788420 TI - Construction of new forms of pyruvate carboxylase to assess the allosteric regulation by acetyl-CoA. AB - The single polypeptide chain of Bacillus thermodenitrificans pyruvate carboxylase (PC) is composed of the biotin carboxylase (BC), carboxyl transferase (CT) and biotin carboxyl carrier protein (BCCP) domains from the amino terminus. This polypeptide chain was divided into two between the CT and BCCP domains. The resulting proteins, PC-(BC + CT) and PC-(BCCP), were expressed in Escherichia coli separately, purified to homogeneity and characterized. PC-(BC + CT) was 4% as active as native PC in the carboxylation of pyruvate with PC-(BCCP) as substrate with a K(m) of 39 microM. Moreover, acetyl-CoA stimulated the carboxylation of PC-(BCCP) about 3-fold, whereas it was without effect in the corresponding reaction with free biotin. In addition to these engineered proteins, another form of enzyme was also constructed in which the BC domain of B.thermodenitrificans PC was replaced with the BC subunit of Aquifex aeolicus PC, whose activity is independent of acetyl-CoA. The resulting chimera was about 7% as active as native PC, but its activity was independent of acetyl-CoA. On the basis of these observations, the mechanism by which acetyl-CoA regulates the reaction of PC is discussed. PMID- 15788421 TI - Redesigning an integral membrane K+ channel into a soluble protein. AB - Even though the structure determination of soluble proteins has become routine, the number of unrelated integral membrane protein structures remains at a few dozen. The importance of this class of proteins to the molecular mechanisms underlying numerous biological phenomena demands that novel experimental techniques be developed to overcome the limitations imposed by conventional detergent-dependent approaches. Here we report the re-engineering of a putative K+ channel protein of unknown structure into a water-soluble analogue. By analyzing evolutionary conservation patterns of related sequences, lipid-facing residues of the primitive channel were identified and mutagenized into more polar alternatives. Further stabilization of the resultant construct was achieved through fusion with maltose-binding protein. The final soluble protein forms a tetramer, suggesting that it accurately models its predecessor. This methodology, as a viable alternative to the use of detergents, should be applicable to a wide range of integral membrane protein families including transporters and other signal transducers. PMID- 15788422 TI - Protein sequence entropy is closely related to packing density and hydrophobicity. AB - We investigated the correlation between the Shannon information entropy, 'sequence entropy', with respect to the local flexibility of native globular proteins as described by inverse packing density. These are determined at each residue position for a total set of 130 query proteins, where sequence entropies are calculated from each set of aligned residues. For the accompanying aggregate set of 130 alignments, a strong linear correlation is observed between the calculated sequence entropy and the corresponding inverse packing density determined at an associated residue position. This region of linearity spans the range of C(alpha) packing densities from 12 to 25 amino acids within a sphere of 9 angstrom radius. Three different hydrophobicity scales all mimic the behavior of the sequence entropies. This confirms the idea that the ability to accommodate mutations is strongly dependent on the available space and on the propensity for each amino acid type to be buried. Future applications of these types of methods may prove useful in identifying both core and flexible residues within a protein. PMID- 15788423 TI - Directed evolution of Pseudomonas aeruginosa lipase for improved amide hydrolyzing activity. AB - A lipase from Pseudomonas aeruginosa was subjected to directed molecular evolution for increased amide-hydrolyzing (amidase) activity. A single round of random mutagenesis followed by screening for hydrolytic activity for oleoyl 2 naphthylamide as compared with that for oleoyl 2-naphthyl ester identified five mutants with 1.7-2.0-fold increased relative amidase activities. Three mutational sites (F207S, A213D and F265L) were found to affect the amidase/esterase activity ratios. The combination of these mutations further improved the amidase activity. Active-site titration using a fluorescent phosphonic acid ester allowed the molecular activities for the amide and the ester to be determined for each mutant without purification of the lipase. A double mutant F207S/A213D gave the highest molecular activity of 1.1 min(-1) for the amide, corresponding to a 2-fold increase compared with that of the wild-type lipase. A structural model of the lipase indicated that the mutations occurred at the sites near the surface and remote from the catalytic triad, but close to the calcium binding site. This study is a first step towards understanding why lipases do not hydrolyze amides despite the similarities to serine proteases in the active site structure and the reaction mechanism and towards the preparation of a general acyl transfer catalyst for the biotransformation of amides. PMID- 15788424 TI - Stromal over-reduction by high-light stress as measured by decreases in P700 oxidation by far-red light and its physiological relevance. AB - The oxidation level of P700 induced by far-red light (DeltaA(FR)) in briefly dark treated leaves of some sun plants decreased during the daytime and recovered at night. The dark recovery of decreased DeltaA(FR) proceeded slowly, with a half time of about 5 h. We propose that stromal over-reduction induced by sunlight was the direct cause of the depression of DeltaA(FR). The depression of DeltaA(FR) found during the daytime was reproduced by controlled illumination with saturating light of fully dark-treated leaves. Simultaneous measurement of P700 redox and chlorophyll fluorescence showed that the depression of DeltaA(FR) was associated with dark reduction of the plastoquinone pool, which represented cyclic electron transport activity. The decrease of DeltaA(FR) in the light stressed chloroplasts was partly reversed by treatment with 2,5-dibromo-3-methyl 6-isopropyl-p-benzoquinone, an inhibitor of electron transport at the cytochrome b6/f complex, and the subsequent addition of methyl viologen, an efficient electron acceptor from photosystem I (PSI), stimulated further recovery, showing that both cyclic electron flow around PSI and the charge recombination within PSI were responsible for the light-induced depression of DeltaA(FR). The dark level of blue-green fluorescence, an indicator of NAD(P)H concentration, from intact chloroplasts was increased by high-light stress, suggesting that NADPH accumulated in stroma as a result of the high-light treatment. Possible effects on photosynthetic activity of over-reduction and its physiological relevance are discussed. PMID- 15788425 TI - Survival of endothelial cells in vitro on Paclitaxel-loaded coronary stents. AB - Coronary stents that are developed for use with balloon angioplasty are known to cause acute occlusion and long-term stenosis. It is likely that a controlled release of drugs at the site of stent implantation might inhibit the proliferation of vascular smooth muscle cells (VSMC) and reduce restenosis. However, if the drug is necrotic and affects cell survival near the implant, it may interrupt the local tissue regeneration. Different methods have been used for the immobilization of drugs with stents to get an effective concentration that inhibits cell proliferation. The objective of this study is to assess the effectiveness of Paclitaxel-loaded stents by immobilization with a biodegradable polymer, to inhibit cell proliferation. The cells used for the evaluation are human umbilical vein endothelial cells (HUVEC) and the proliferation rate of these cells on the drug-coated stent is compared against an uncoated stent for a 72-h period. Evaluations were also made to differentiate between cell apoptosis and necrosis to prove that the drug released is not deleterious to the surrounding tissue. While a similar initial cell adhesion is observed in bare and coated stents, the proliferation of HUVEC is negligible when grown on a drug coated stent (p < 0.001). By specific staining techniques, the cells on the drug coated stents are found to be apoptotic and not necrotic, throughout the evaluation period. In vitro leukocyte adhesion and platelet deposition on the drug-coated stents are found to be low when they are exposed to human blood and platelet-rich plasma (PRP), suggesting that the coated stents may not be thrombogenic in vivo. Therefore, drug coating of stents using the described technique may have a considerable promise for the prevention of neointimal proliferation, restenosis, and associated failure of angioplasty. PMID- 15788426 TI - Synthesis and characterization of polyamide of trans-4-hydroxy-L-proline used as porogen filler in acrylic bone cement. AB - The aim of this study was to synthesize on a larger scale, an experimental polyamide based on an amino acid of trans-4-hydroxy-L-proline. The polyamide of trans-4-hydroxy-L-proline has been used as porogen filler (i.e., a hydrophilic pore generating material) in nondegradable acrylic bone cement. In in vitro studies, this hydrophilic filling component has been shown to form porosity within the acrylic bone cement in an aqueous environment. The formation of in situ porosity in the acrylic polymer matrix is believed to improve the fixation between the cement and the living bone. Namely, a porous structure can support bone ingrowth and strengthen the mechanical connection between the acrylic bone cement and the bone. The monomer, trans-4-hydroxy-L-proline methyl ester, was prepared from trans-4-hydroxy-L-proline by means of two steps, and the monomer was then polymerized to polyamide of trans-4-hydroxy-L-proline. The polymerization was carried out using a melt polycondensation method. The molecular weights (M(psi)) of the produced polyamides were between 1800 and 3600. The products were characterized by FTIR and (1)H-NMR spectroscopy. PMID- 15788427 TI - Porous poly (DL-lactic acid) modified chitosan-gelatin scaffolds for tissue engineering. AB - Chitosan-gelatin (Cs-Gel) scaffolds are modified with poly (DL-lactic acid) (PDLLA) dichloromethane solution of different concentrations (0.1, 0.5, and 1.0%) and immersed in water after the evaporation of the solvent. The swollen scaffolds are freeze dried. The concentration of PDLLA has significant effects on both the physicomechanical properties and the cytocompatibility. Data reveal that only the 0.1% concentration could increase the tensile strength fourfold in comparison with the pristine Cs-Gel scaffold, while maintaining the human fibroblast adhesion, migration, and proliferation just like the Cs-Gel scaffold. PMID- 15788428 TI - Biocompatibility of Poly(epsilon-caprolactone) scaffold modified by chitosan--the fibroblasts proliferation in vitro. AB - In this study, the surface of poly(epsilon-caprolactone) (PCL) scaffold was modified by chitosan (CS) in order to enhance its cell affinity and biocompatibility. It is demonstrated by scanning electronic microscopy (SEM) that when 0.5-2.0 wt% chitosan solutions are used to modify the PCL scaffold, the amount of adhesion of the fibroblasts on the chitosan-modified PCL scaffolds dramatically increase when compared to the control after 7 days cell culture. The results indicate that the chitosan-modified PCL scaffolds are more favorable for cell proliferation by improving the scaffold biocompatibility. The improvement may be helpful for the extensive applications of PCL scaffold in heart valve and blood vessel tissue engineering. PMID- 15788429 TI - Bone in-growth induced by biphasic calcium phosphate ceramic in femoral defect of dogs. AB - Biphasic calcium phosphate (BCP) ceramics consisting of hydroxyapatite (HA) and tricalcium phosphate (TCP) has been used as a bone graft material during the last decade. In this paper, we report the bone in-growth induced by BCP ceramic in the experimentally created circular defects in the femur of dogs. This BCP ceramic consists of 55% hydroxyapatite (HA) and 45% b-tricalcium phosphate (TCP) prepared in situ by the microwave method. The defects were created as 4-mm holes on the lateral aspect of the femur of dogs and the holes were packed with the implant material. The defective sites were radiographed at a period of 4, 8, and 12 weeks postoperatively. The radiographical results showed that the process of ossification started after 4 weeks and the defect was completely filled with new woven bone after 12 weeks. Histological examination of the tissue showed the formation of osteoblast inducing the osteogenesis in the defect. The collageneous fibrous matrix and the complete Haversian system were observed after 12 weeks. The blood serum was collected postoperatively and biochemical assays for alkaline phosphatase activity were carried out. The measurement of alkaline phosphatase activity levels also correlated with the formation of osteoblast-like cells. This microwave-prepared BCP ceramic has proved to be a good biocompatible implant as well as osteoconductive and osteoinductive materials to fill bone defects. PMID- 15788430 TI - Parsons revisited: from the sick role to...? AB - This article revisits Parsons' insights on medicine, health and illness in the light of contemporary debates in medical sociology and beyond. A preliminary balance sheet of the Parsonian legacy is first provided, taking on board standard accounts and criticisms of Parsons' work within medical sociology to date. The remainder of the article, in contrast, involves a close re-reading of Parsons in the light of contemporary sociological debates on the body, emotion, trust, uncertainty and health, including late modern and postmodern interpretations of his work. Parsons, it is concluded, despite his (many) critics and detractors, has much to contribute here, not simply in terms of past insights, but also with regard to the present and future. PMID- 15788431 TI - Radical contextualization: contributions to an anthropology of racial/ethnic health disparities. AB - There is indisputable evidence of deep and persistent racial/ethnic inequalities in health status and health care in the USA. Growing awareness of these disparities has fueled a cross-disciplinary debate about appropriate approaches to racial/ethnic disparities in public health research and policy discourse, yet anthropologists have been marginalized in this discourse. What does the current work of anthropologists have to offer that is most useful in the crucial work of understanding and eliminating health disparities? We examine anthropological research and practice that constitute core contributions to an anthropology of racial/ethnic health disparities. We identify the following themes: (1) using ethnography as a tool for new inequality knowledge; (2) studying up; and (3) formulating alternative models of biosocial pathogenesis. These elements of anthropological methods, theory and practice can contribute to a better understanding of the social processes that underpin racial/ethnic health disparities and help identify opportunities for interrupting them. PMID- 15788432 TI - Beyond the mask: women's experiences of public and private ageing during midlife and their use of age-resisting activities. AB - Accounts of ageing often employ the metaphor of a mask and suggest that individuals are motivated to present a youthful image. Drawing on interview data, we reveal that women aged 51-57 years distinguish between what we call 'public' and 'private' body ageing, both of which have an impact on age-resistance. Public ageing is visible, arising from physical changes in body appearance. These changes have the potential for concealment through age-resisting activities. Private ageing is less visible and arises largely from physiological changes within the body, which were perceived by women as irreversible indicators of ageing. This obduracy of the body led women to perceive themselves as ageing and also deterred them from participating in age-resistance. In contrast to masking theories, our study shows that most women in their 50s wanted to project a socially acceptable image that reflected their subjective sense of growing old. PMID- 15788433 TI - Diagnosing the male steroid user: drug use, body image and disordered masculinity. AB - As steroid use has gained prominence as a dangerous form of substance abuse, two main sets of discourses have been deployed to investigate and ameliorate this emerging public health threat. This article examines these two discursive frameworks and their constitution of the male steroid user as psychologically disordered, drawing on a range of medical and psychological literature. The first framework understands steroid use as a form of illicit drug use, and constitutes the steroid user as an antisocial and excessively masculine subject. The second locates steroid use within the field of body image disorder, producing the steroid user as a damaged and feminized male, a vivid example of masculinity in crisis. Both of these approaches tend to elide the specificity of steroid use and its associated bodily practices in their eagerness to form it into an easily comprehended entity which can be targeted by medical and legal governance. PMID- 15788434 TI - Understanding variations in exposure to social stress. AB - Although exposure to stress is a central component of the stress process paradigm, little research has explicitly sought to identify antecedents of stress exposure. Based on a probability sample of 1393 adults aged 18-55 residing in Toronto, Canada, this research examines the effects of social status, past adversity, social and personal resources and history of mental disorder on recent exposure to stress. Consistent with earlier findings, results indicate that younger adults, divorced individuals and those with lower socioeconomic status, experience greater levels of social stress. Although respondent's history of major depressive disorder was related to all types of stress exposure, past cumulative adversity was the most powerful predictor of both total current stress (operant burden) and the subsequent onset of life events, independent of other antecedents. Findings suggest that the onset of chronic stress is more affected by personal characteristics, such as emotional reliance and disorder history, than is onset of life events. Implications of these findings are discussed. PMID- 15788435 TI - Reconstituting populations through evidence-based medicine: an ethnographic account of recommending procedures for diagnosing type 2 diabetes in clinical practice guidelines. AB - This ethnographic investigation responds to calls for more social science research on -- versus for or against -- evidence-based medicine (EBM). It centers on the recommendations endorsed, in a set of clinical practice guidelines, by Canadian specialists in the late 1990s for diagnosing diabetes. Empirically, the article mainly relies on public presentations and discussion at the Canadian Diabetes Association Professional Conferences (1997-2001), supplemented by findings from documentary sources, direct observation, participant-observation and interviews. It confirms the importance of clinical reason and clinical epidemiology as preconditions for EBM, the textually mediated character of EBM and patients' bodies as sites for producing knowledge crucial to EBM. Most significantly, it also demonstrates the importance of non-patients' bodies as sites for producing knowledge that is crucial to EBM and its politics. EBM politics encompass the discursive, socio-technical and visceral reconstitution of populations. PMID- 15788436 TI - Floral ontogeny in Scirpus, Eriophorum and Dulichium (Cyperaceae), with special reference to the perianth. AB - BACKGROUND AND AIMS: Based on molecular phylogenetic analysis, it has been suggested recently that the Cyperaceae comprises only two subfamilies: the Mapanioideae and the Cyperoideae. In most flowers of the Cyperoideae, the whorl of inner stamens is reduced, resulting in tetracyclic flowers. In the more primitive (scirpoid) genera within the Cyperoideae, the perianth consists of two polysymmetric whorls, whereas the perianth parts in the more derived genera have been subject to modifications and/or reduction. Comparative studies of the many silky hairs of Eriophorum and of the eight bristles of Dulichium have given rise to much discussion about their homology. METHODS: The spikelet and floral ontogeny in freshly collected inflorescences was investigated using scanning electron microscopy. KEY RESULTS: Complete floral ontogenies are presented for Scirpus sylvaticus L., Eriophorum latifolium Hoppe and Dulichium arundinaceum (L.) Britton, with special reference to the perianth. The results in S. sylvaticus confirm the trimerous monocot-like organization of the flower. It is used as a model for floral development in Cyperoideae. In the early developmental stages, the androecium of E. latifolium is surrounded by a massive perigonial primordium, from which the many hair-like bristles originate. Consequently, the stamens develop among the hair primordia, more or less simultaneously. The hairs are arranged in whorls, which develop centripetally. The development of the perianth in D. arundinaceum starts with the formation of three initial perianth primordia opposite the stamens. Subsequently, two more abaxial bristle primordia, alternating with the stamens, originate simultaneously with the appearance of three adaxial bristle primordia in the zone where an adaxial inner perianth primordium is expected. CONCLUSIONS: The floral development in E. latifolium and D. arundinaceum can be considered as variations upon the scirpoid floral ontogenetic theme. PMID- 15788437 TI - Enhanced external counterpulsation: a new technique to augment renal function in liver cirrhosis. AB - BACKGROUND: Advanced liver cirrhosis is characterized by cardiovascular changes, such as low arterial blood pressure, peripheral vasodilation and renal vasoconstriction. As a consequence, renal hypoperfusion, impaired diuresis and natriuresis and eventual hepatorenal syndrome may ensue. Previous studies using head-out water immersion to increase central blood volume have demonstrated the functional nature of the renal abnormalities. Enhanced external counterpulsation (EECP) is a new non-invasive cardiac assist device to augment diastolic blood pressure by electrocardiogram-triggered diastolic inflation and deflation of cuffs wrapped around the lower extremities. We investigated whether EECP would improve renal dysfunction of liver cirrhosis. METHODS: Twelve healthy controls and 19 patients with liver cirrhosis were observed during 2 h of baseline followed by 2 h of EECP. The following parameters of renal and cardiovascular function were measured: renal plasma flow by para-aminohippurate clearance, glomerular filtration rate (GFR) by inulin clearance, urine flow rate, urinary excretion rates of sodium and chloride, mean arterial blood pressure (MAP), renal vascular resistance (RVR) and plasma concentrations of renin, atrial natriuretic peptide (ANP), endothelin-1, antidiuretic hormone, epinephrine and N-epinephrine. RESULTS: EECP was well tolerated by healthy controls and cirrhotic patients alike. EECP increased MAP (cirrhotic patients: from 74+/-18 to 88+/-20 mmHg, P<0.01; controls: from 89+/-8 to 94+/-5 mmHg, P = NS) and ANP (cirrhotic patients: from 23+/-18 to 30+/-20 ng/l, P<0.05; controls: from 11+/-4 to 16+/-5 ng/l, P<0.01). The plasma renin concentration decreased (cirrhotic patients: from 98+/-98 to 58+/-57 ng/l, P<0.01; controls: from 4.6+/-1.6 to 3.4+/-1.1 ng/l, P<0.01). This was associated with improvement of the urinary flow rate (cirrhotic patients: from 3.6+/-1.8 to 4.6+/-0.7 ml/min, P<0.05; controls: from 1.8+/-1.5 to 2.8+/-1.9 ml/min, P<0.05), as well as of the sodium and chloride excretion rates in both groups. However, in contrast to healthy controls, GFR and renal plasma flow in cirrhotic patients failed to rise significantly. Renal vascular resistance fell numerically in healthy controls (68+/-5 vs 55+/-4 mmHg . min/l; P = NS). In contrast, RVR showed a significant increase by approximately 20% in cirrhosis (67+/-4 vs 80+/-8 mmHg . min/l; P<0.05). Endothelin-1 levels fell in controls (0.38+/-0.42 vs 0.31+/-0.35; P<0.05), whereas they remained statistically unchanged in cirrhotic patients. Epinephrine, N-epinephrine and vasopressin were not altered by EECP in either group. CONCLUSIONS: EECP is an effective procedure to augment renal excretory function in healthy volunteers as well as in patients with cirrhosis. In healthy volunteers, GFR and renal plasma flow increased during EECP. In contrast, these parameters remained unchanged in the patients and their renal vascular resistance increased during EECP. Therefore, EECP improves diuresis, but does not influence the vasoconstrictive dysregulation of the kidneys in liver cirrhosis. PMID- 15788438 TI - Renal and thymic pathology in thymoma-associated nephropathy: report of 21 cases and review of the literature. AB - BACKGROUND: Acquired thymic disease (malignant thymoma or thymic hyperplasia) is associated with various autoimmune diseases, such as myasthenia gravis (MG), pure red-cell aplasia (PRCA), pemphigus vulgaris or systemic lupus erythematosus (SLE). Renal disease has rarely been observed in association with thymoma. METHODS: This retrospective, multicentric study collected data on patients with thymic disease and biopsy-proven renal involvement. RESULTS: Twenty-one patients were studied (age: 49+/-14 years; male/female ratio: 8/13). Thymic pathology revealed mostly high-grade malignant thymoma (B2 and AB type); two cases were associated with non-malignant thymic hyperplasia. MG was found in nine out of 21 cases, SLE in three, PRCA in three and pemphigus in two. In 47% of these cases, nephropathy occurred after curative treatment of thymoma (108+/-83 months; range: 8-180 months), mainly based on surgical thymectomy associated with radiotherapy. Clinical and laboratory findings included nephrotic syndrome (75%), renal failure (50%), frequent presence of antinuclear antibodies and hypogammaglobulinaemia. Renal pathology showed minimal change disease in 14 patients and focal segmental glomerulosclerosis (FSGS) in one. Membranous nephropathy was observed in four cases, ANCA-associated glomerulonephritis in two and thrombotic microangiopathy in one. Most patients with minimal change disease or FSGS (11/13) were steroid sensitive. Despite good response to steroids, 38% of patients died from thymoma and 17% developed end-stage renal failure. CONCLUSIONS: Glomerulopathy can be associated with thymoma or thymic hyperplasia. The present series shows that minimal change disease is the most frequent thymoma-associated glomerular lesion and that it may occur several years after thymectomy. PMID- 15788439 TI - Diabetes-prone NOD mice show an expanded subpopulation of mature circulating monocytes, which preferentially develop into macrophage-like cells in vitro. AB - In the nonobese diabetic (NOD) mouse, a model of autoimmune diabetes, dendritic cells (DC), and macrophages (Mphi) is important for the initiation and progression of autoimmunity and the final destruction of beta-cells. Previous studies suggested that an aberrant development of DC and Mphi is related to their pathogenic function. To study this in vivo, we investigated NOD mouse monocytes, the direct precursors of DC, and Mphi. The recently described discrimination between immature (Ly-6C(high)) and mature (Ly-6C(low)) monocytes enabled us to investigate the apportioning between blood monocyte populations in the NOD mouse, which had an abnormally high number of mature monocytes in circulation, and this phenomenon appeared to be intrinsic to the NOD background, as nonobese resistant (NOR) and NOD-H2b mice also showed this altered balance. After depletion by apoptosis-inducing liposomes, the reappearance and transition of immature-to mature monocytes had similar kinetics as control mice but led again to the presence of a larger, mature monocyte compartment in the blood. In addition, although monocytes from C57BL mice down-regulated their capability to adhere to fibronectin and intercellular adhesion molecule-1 upon maturation, the mature NOD monocytes retained their high adhesion capacity, characteristic of immature cells. Furthermore, both monocyte subpopulations of NOD mice showed enhanced differentiation into Mphi-like F4/80(high) cells in vitro. In conclusion, mice with the NOD background have raised numbers of mature monocytes in the circulation and a proinflammatory, Mphi-directed monocyte development. PMID- 15788440 TI - CTLA4-CD80/CD86 interactions on primary mouse CD4+ T cells integrate signal strength information to modulate activation with Concanavalin A. AB - The mechanisms by which concanavalin A (Con A), a lectin, activates T cells are poorly studied. A low dose of Con A is stimulatory for T cells, whereas a high dose of Con A results in suppression of proliferation and enhanced T cell death. The expression and functional roles of costimulatory receptors, CD28 and cytotoxic T-lymphocyte antigen 4 (CTLA4), and their ligands, CD80 and CD86, on primary mouse CD4(+) T cells after activation with different doses of Con A were studied. CTLA4-CD80/CD86 interactions in this T:T cell activation model demonstrate distinct outcomes depending on the dose of Con A. CTLA4-CD80/CD86 interactions inhibit CD4(+) T cell cycling and survival after activation with a suppressive dose of Con A by increasing oxidative stress and decreasing levels of BclX(L). The enhanced CD4(+) T cell death with a suppressive dose of Con A is dependent on excess H(2)O(2) and nitric oxide but is independent of Fas and caspase activity. It is surprising that the increased proliferation of CD4(+) T cells with a suppressive dose of Con A on blocking CTLA4-CD80/CD86 interactions is largely interleukin (IL)-2-independent but is cyclosporine A-sensitive. On activation with a stimulatory dose of Con A, CTLA4-CD80/CD86 interactions enhance T cell activation and survival by reducing the production of reactive oxygen species, increasing IL-2 and BclX(L) levels. Here IL-10 but not transforming growth factor-beta plays a functional role. In summary, CTLA4-CD80/CD86 interactions on T cells integrate signal strength, based on the dose of Con A, to enhance or inhibit primary mouse CD4(+) T cell cycling and survival. PMID- 15788441 TI - Regulated expression of platelet factor 4 in human monocytes--role of PARs as a quantitatively important monocyte activation pathway. AB - Human mononuclear phagocytes have recently been shown to express constitutively and even more so, upon stimulation with bacteria, fungi, lipopolysaccharide (LPS), zymosan, or thrombin platelet basic protein (PBP). This CXC chemokine as well as platelet factor 4 (PF4), which is located genomically at a short distance from the PBP, were previously considered to be specific markers for the megakaryocyte cell lineage. Both chemokines have signaling and antimicrobial activity. In the present studies, transcriptional and expressional regulation of PF4 and related chemokines was studied in human monocytes. As shown by quantitative mRNA analysis, Western blots, radioimmunoprecipitation of cell extracts, and immunofluorescence and quantitatively with enzyme-linked immunosorbent assay, human monocytes express PF4 in the same order of magnitude as the known, regulated CXC chemokine interleukin (IL)-8. Expression of PF4 is up regulated at the mRNA and protein level by thrombin and mediated by proteinase activated receptors (PARs), resulting in a 32- to 128-fold higher mRNA level and leading to an up-to-sixfold increase of the peptide concentration in monocyte culture supernatants. Thrombin and the synthetic ligand of PAR-1 and PAR-2, SFLLRN, also induced comparable increases in the levels of mRNA for PBP, IL-8, regulated on activation, normal T expressed and secreted (RANTES), monocyte chemoattractant protein-1, and macrophage-inflammatory protein-1alpha and increased synthesis of these chemokines as shown by immunofluorescence or a quantitative immunobead-based method. The induction of increased mRNA levels for all chemokines by SFLLRN was unsurpassed by LPS, zymosan, interferon-gamma (IFN gamma), tumor necrosis factor alpha (TNF-alpha), and IL-1. Activation of monocytes through PARs represents an alternate activation mechanism, independent from IFN-gamma, TNF-alpha, or other signaling pathways. PMID- 15788442 TI - The effects of capecitabine in Raynaud's disease: a case report. PMID- 15788443 TI - Can bleomycin toxicity in the treatment of testis cancer be batch related? PMID- 15788444 TI - Is high-dose chemotherapy with peripheral stem cell rescue a suitable option for elderly patients affected by aggressive non-Hodgkin's lymphoma? PMID- 15788445 TI - The vasoactive peptide adrenomedullin is secreted by adipocytes and inhibits lipolysis through NO-mediated beta-adrenergic agonist oxidation. AB - Adipocytes are known to secrete a number of adipokines, but many adipocyte secretions and their functional importance remain to be characterized. This work shows that human white adipocytes and 3T3-F442A-derived adipocytes produce adrenomedullin (AM) and that AM acts in an autocrine/paracrine way on lipid metabolism by extracellular inactivation of isoproterenol, a beta-adrenergic agonist. AM is described as a counter-regulatory factor involved in the control of cardiovascular homeostasis. This peptide is believed to protect the heart from several complications implicated in obesity-linked cardiomorbidity, such as arterial hypertension, cardiac fibrosis, and decreased sinusal variability. The exact source of circulating AM remains a matter of debate, although endothelial and vascular smooth muscle cells seem to be important sites of production. We show that human adipose cells and 3T3-F442A-derived adipocytes express AM receptors and secrete AM. The function of this feature was investigated in 3T3 F442A cell line at the level of lipolysis regulation. AM inhibited beta adrenergic-stimulated lipolysis by a nitric oxide (NO)-dependent mechanism, inducing a significant decrease in pD2 value for isoproterenol (8.6 +/- 0.2 vs. 9.8 +/- 0.1, P<0.001). This effect is cGMP-independent since it occurred in the presence of the NO-sensitive guanylate cyclase inhibitor ODQ. It is apparently mediated by a novel extracellular mechanism. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) demonstrated that AM-produced NO oxidized isoproterenol to generate its aminochrome, namely isoprenochrome. Isoprenochrome amounts were increased 3.62 +/- 1.13-fold in cell culture media (P<0.05). We describe for the first time that AM down-regulates lipolysis in adipocytes through the chemical modification of a beta-agonist. PMID- 15788446 TI - Assessment of a dual regulatory role for NO in liver regeneration after partial hepatectomy: protection against apoptosis and retardation of hepatocyte proliferation. AB - The role of hepatic nitric oxide (NO) in liver regeneration after partial hepatectomy (PH) was studied in animals carrying a nitric oxide synthase-2 transgene under the control of the phospho(enol)pyruvate carboxykinase promoter. These mice expressed NOS-2 in liver cells under fasting conditions. Liver mass recovery and molecular parameters related to cell proliferation were determined after PH. Preexisting hepatic NO synthesis, as well as NO delivery by NO-donors, impaired early signaling (for example, attenuated NF-kappaB activation and TNF alpha and IL-6 release). The regenerative process was also impaired as a result of an insufficient proliferative response, but mouse survival after surgery was not compromised. However, NO exerted a protective role against apoptosis in transgenic hepatectomized mice. Local production of NO in liver cells, achieved by hydrodynamic-based transfection with a NOS-2-encoding plasmid, also resulted in delayed liver recovery after PH and also protected against Fas-mediated apoptosis. These data show that sustained presence of NO after PH exerts a dual role: attenuating liver regeneration while efficiently protecting against liver apoptosis. PMID- 15788447 TI - Hepatic leptin signaling in obesity. AB - Obesity, a state of apparent "leptin resistance" is well known to be associated with insulin resistance. In diet-induced obesity (DIO), hepatic insulin signaling is impaired but the link between leptin and insulin signaling pathways is only incompletely defined. The aim of the present study was to evaluate the effects of DIO on leptin and insulin cross-signaling in the liver. Leptin receptor expression was measured by in situ hybridization with pan-leptin receptor probes and by immunoblotting. Furthermore, intracellular signaling was investigated in vivo under basal conditions and at 45 and 360 min after stimulation with a bolus of human recombinant leptin (hrec-leptin; 1 mg/kg body wt) or saline. At baseline, all forms of the leptin receptor were markedly to completely down regulated in DIO rats. Hrec-leptin bolus injection stimulated leptin-dependent signaling with a fivefold increase in JAK-2pY in lean but not in DIO rats. Basal IRpY, IRS-1pY, IRS-1p85, IRS-2pY, IRSp85, and PKBpT308 levels were reduced (P<0.01) in DIO rats as compared with lean controls. Basal GSK-3beta serine phosphorylation (S9) was higher (P<0.01) in lean animals along with lower basal PEPCK activity compared with DIO rats consistent with the insulin and leptin resistance of the latter. Only in lean animals phosphorylation of PKB (T308) and GSK-3beta (S9) was acutely stimulated by leptin at 45 min followed by inhibition at 6 h after application. AMPKalpha protein levels as well as basal and leptin stimulated total and alpha-specific AMPK activity were comparable in both groups. These data show that in a model of dietary-induced obesity 1) leptin receptors and subsequent signaling events are down-regulated, 2) basal insulin signaling is impaired, and 3) the cross-talk between leptin and insulin signaling is differentially regulated by the nutritional status, which is sensed by AMPK in rat liver. Thus, the liver seems to play a major role in the modulation of the leptin signal and insulin resistance in obesity. PMID- 15788448 TI - PPAR-gamma modulates allergic inflammation through up-regulation of PTEN. AB - The ligand-activated nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) has been shown to regulate cell activation, differentiation, proliferation, and/or apoptosis. PPARgamma is also associated with anti inflammatory responses. However, the signaling mechanism remains elusive. We have used a mouse model for asthma to determine the effect of PPARgamma agonists, rosiglitazone or pioglitazone, and PPARgamma on allergen-induced bronchial inflammation and airway hyperresponsiveness. Administration of PPARgamma agonists or adenovirus carrying PPARgamma cDNA (AdPPARgamma) reduced bronchial inflammation and airway hyperresponsiveness. Expression of PPARgamma was increased by ovalbumin (OVA) inhalation, and the increase was further enhanced by the administration of the PPARgamma agonists or AdPPARgamma. Levels of IL-4, IL 5, IL-13, and eosinophil cationic protein were increased after OVA inhalation, and the increased levels were significantly reduced by the administration of PPARgamma agonists or AdPPARgamma. The results also showed that the administration of PPARgamma agonists or AdPPARgamma up-regulated phosphatase and tensin homologue deleted on chromosome ten (PTEN) expression in allergen-induced asthmatic lungs. This up-regulation correlated with decreased phosphatidylinositol 3-kinase activity as measured by reduced phosphorylation of Akt. These findings demonstrate a protective role of PPARgamma in the pathogenesis of the asthma phenotype through regulation of PTEN expression. PMID- 15788449 TI - Measuring protein-protein interactions inside living cells using single color fluorescence correlation spectroscopy. Application to human immunodeficiency virus type 1 integrase and LEDGF/p75. AB - Recently we described the interaction of human immunodeficiency virus type 1 (HIV 1)1 integrase (IN) with a cellular protein, lens epithelium-derived growth factor/transcription co-activator p75 (LEDGF/p75). We now present the study of the diffusion behavior of the three independent domains of IN and LEDGF/p75 using fluorescence correlation microscopy (FCM). We show that diffusion in the cell of the different enhanced green fluorescent protein (EGFP) fusion proteins is described by two components with different fractions and that the average parameters in the nucleus are comparable with those in the cytoplasm. In addition, we demonstrate that specific interaction between EGFP-fused HIV-1 IN and LEDGF/p75 results in a shift in diffusion coefficient (D). The opposite shift was observed in an IN-deletion mutant that does not exhibit LEDGF/p75 binding or in a LEDGF/p75 knock-down experiment using siRNA. We thus demonstrate that protein-protein interactions can be studied in living cells, using single-color FCM (scFCM). PMID- 15788450 TI - even-skipped is not a pair-rule gene but has segmental and gap-like functions in Oncopeltus fasciatus, an intermediate germband insect. AB - The pair-rule gene even-skipped is required for the initiation of metameric pattern in Drosophila. But Drosophila segmentation is evolutionarily derived and is not representative of most insects. Therefore, in order to shed light on the evolution of insect segmentation, homologs of the pair-rule gene even-skipped have been studied in several insect taxa. However, most of these studies have reported the expression eve but not its function. We report the isolation, expression and function of the homolog of Drosophila even-skipped from the intermediate germband insect Oncopeltus fasciatus. We find that in Oncopeltus, even-skipped striped expression initiates in a segmental and not pair-rule pattern. Weak RNAi suppression of Oncopeltus even-skipped shows no apparent pair rule like phenotype, while stronger RNAi suppression shows deletion of nearly the entire body. These results suggest that in Oncopeltus, even-skipped is not acting as a pair-rule gene. In almost all insects, prior to its striped expression, even skipped is expressed in a conserved broad gap-like domain but its function has been largely ignored. We find that this early broad domain is required for activation of the gap genes hunchback and Kruppel. Given the large RNAi deletion phenotype and its regulation of hunchback and Kruppel, even-skipped seems to act as an uber-gap gene in Oncopeltus, indicating that it may have both upstream and downstream roles in segmentation. PMID- 15788451 TI - Applications of mRNA injections for analyzing cell lineage and asymmetric cell divisions during segmentation in the leech Helobdella robusta. AB - Synthetic mRNAs can be injected to achieve transient gene expression even for 'non-model' organisms in which genetic approaches are not feasible. Here, we have used this technique to express proteins that can serve as lineage tracers or reporters of cellular events in embryos of the glossiphoniid leech Helobdella robusta (phylum Annelida). As representatives of the proposed super-phylum Lophotrochozoa, glossiphoniid leeches are of interest for developmental and evolutionary comparisons. Their embryos are suitable for microinjection, but no genetic approaches are currently available. We have injected segmentation stem cells (teloblasts) with mRNAs encoding nuclear localized green fluorescent protein (nGFP) and its spectral variants, and have used tandem injections of nGFP mRNA followed by antisense morpholino oligomer (AS MO), to label single blast cell clones. These techniques permit high resolution cell lineage tracing in living embryos. We have applied them to the primary neurogenic (N) lineage, in which alternate segmental founder cells (nf and ns blast cells) contribute distinct sets of progeny to the segmental ganglia. The nf and ns blast cell clones exhibit strikingly different cell division patterns: the increase in cell number within the nf clone is roughly linear, while that in the ns clone is almost exponential. To analyze spindle dynamics in the asymmetric divisions of individual blast cells, we have injected teloblasts with mRNA encoding a tau::GFP fusion protein. Our results show that the asymmetric divisions of n blast cells result from a posterior shift of both the spindle within the cell and the midbody within the mitotic spindle, with differential regulation of these processes between nf and ns. PMID- 15788452 TI - Cdx2 is required for correct cell fate specification and differentiation of trophectoderm in the mouse blastocyst. AB - Blastocyst formation marks the segregation of the first two cell lineages in the mammalian preimplantation embryo: the inner cell mass (ICM) that will form the embryo proper and the trophectoderm (TE) that gives rise to the trophoblast lineage. Commitment to ICM lineage is attributed to the function of the two transcription factors, Oct4 (encoded by Pou5f1) and Nanog. However, a positive regulator of TE cell fate has not been described. The T-box protein eomesodermin (Eomes) and the caudal-type homeodomain protein Cdx2 are expressed in the TE, and both Eomes and Cdx2 homozygous mutant embryos die around the time of implantation. A block in early TE differentiation occurs in Eomes mutant blastocysts. However, Eomes mutant blastocysts implant, and Cdx2 and Oct4 expression are correctly restricted to the ICM TE. Blastocoel formation initiates in Cdx2 mutants but epithelial integrity is not maintained and embryos fail to implant. Loss of Cdx2 results in failure to downregulate Oct4 and Nanog in outer cells of the blastocyst and subsequent death of those cells. Thus, Cdx2 is essential for segregation of the ICM and TE lineages at the blastocyst stage by ensuring repression of Oct4 and Nanog in the TE. PMID- 15788453 TI - IrreC/rst-mediated cell sorting during Drosophila pupal eye development depends on proper localisation of DE-cadherin. AB - Remodelling of tissues depends on the coordinated regulation of multiple cellular processes, such as cell-cell communication, differential cell adhesion and programmed cell death. During pupal development, interommatidial cells (IOCs) of the Drosophila eye initially form two or three cell rows between individual ommatidia, but then rearrange into a single row of cells. The surplus cells are eliminated by programmed cell death, and the definitive hexagonal array of cells is formed, which is the basis for the regular pattern of ommatidia visible in the adult eye. Here, we show that this cell-sorting process depends on the presence of a continuous belt of the homophilic cell adhesion protein DE-cadherin at the apical end of the IOCs. Elimination of this adhesion belt by mutations in shotgun, which encodes DE-cadherin, or its disruption by overexpression of DE cadherin, the intracellular domain of Crumbs, or by a dominant version of the monomeric GTPase Rho1 prevents localisation of the transmembrane protein IrreC rst to the border between primary pigment cells and IOCs. As a consequence, the IOCs are not properly sorted and supernumerary cells survive. During the sorting process, Notch-mediated signalling in IOCs acts downstream of DE-cadherin to restrict IrreC-rst to this border. The data are discussed in relation to the roles of selective cell adhesion and cell signalling during tissue reorganisation. PMID- 15788454 TI - Ventral migration of early-born neurons requires Dcc and is essential for the projections of primary afferents in the spinal cord. AB - Neuronal migration and lamina-specific primary afferent projections are crucial for establishing spinal cord circuits, but the underlying mechanisms are poorly understood. Here, we report that in mice lacking Dcc (deleted in colorectal cancer), some early-born neurons could not migrate ventrally in the spinal cord. Conversely, forced expression of Dcc caused ventral migration and prevented dorsolateral migration of late-born spinal neurons. In the superficial layer of the spinal cord of Dcc-/- mutants, mislocalized neurons are followed by proprioceptive afferents, while their presence repels nociceptive afferents through Sema3a. Thus, our study has shown that Dcc is a key molecule required for ventral migration of early-born neurons, and that appropriate neuronal migration is a prerequisite for, and coupled to, normal projections of primary afferents in the developing spinal cord. PMID- 15788455 TI - Role of the hindbrain in dorsoventral but not anteroposterior axial specification of the inner ear. AB - An early and crucial event in vertebrate inner ear development is the acquisition of axial identities that in turn dictate the positions of all subsequent inner ear components. Here, we focus on the role of the hindbrain in establishment of inner ear axes and show that axial specification occurs well after otic placode formation in chicken. Anteroposterior (AP) rotation of the hindbrain prior to specification of this axis does not affect the normal AP orientation and morphogenesis of the inner ear. By contrast, reversing the dorsoventral (DV) axis of the hindbrain results in changing the DV axial identity of the inner ear. Expression patterns of several ventrally expressed otic genes such as NeuroD, Lunatic fringe (Lfng) and Six1 are shifted dorsally, whereas the expression pattern of a normally dorsal-specific gene, Gbx2, is abolished. Removing the source of Sonic Hedgehog (SHH) by ablating the floor plate and/or notochord, or inhibiting SHH function using an antibody that blocks SHH bioactivity results in loss of ventral inner ear structures. Our results indicate that SHH, together with other signals from the hindbrain, are important for patterning the ventral axis of the inner ear. Taken together, our studies suggest that tissue(s) other than the hindbrain confer AP axial information whereas signals from the hindbrain are necessary and sufficient for the DV axial patterning of the inner ear. PMID- 15788456 TI - Initial formation of zebrafish brain ventricles occurs independently of circulation and requires the nagie oko and snakehead/atp1a1a.1 gene products. AB - The mechanisms by which the vertebrate brain develops its characteristic three dimensional structure are poorly understood. The brain ventricles are a highly conserved system of cavities that form very early during brain morphogenesis and that are required for normal brain function. We have initiated a study of zebrafish brain ventricle development and show here that the neural tube expands into primary forebrain, midbrain and hindbrain ventricles rapidly, over a 4-hour window during mid-somitogenesis. Circulation is not required for initial ventricle formation, only for later expansion. Cell division rates in the neural tube surrounding the ventricles are higher than between ventricles and, consistently, cell division is required for normal ventricle development. Two zebrafish mutants that do not develop brain ventricles are snakehead and nagie oko. We show that snakehead is allelic to small heart, which has a mutation in the Na+K+ ATPase gene atp1a1a.1. The snakehead neural tube undergoes normal ventricle morphogenesis; however, the ventricles do not inflate, probably owing to impaired ion transport. By contrast, mutants in nagie oko, which was previously shown to encode a MAGUK family protein, fail to undergo ventricle morphogenesis. This correlates with an abnormal brain neuroepithelium, with no clear midline and disrupted junctional protein expression. This study defines three steps that are required for brain ventricle development and that occur independently of circulation: (1) morphogenesis of the neural tube, requiring nok function; (2) lumen inflation requiring atp1a1a.1 function; and (3) localized cell proliferation. We suggest that mechanisms of brain ventricle development are conserved throughout the vertebrates. PMID- 15788457 TI - Non-canonical functions of hunchback in segment patterning of the intermediate germ cricket Gryllus bimaculatus. AB - In short and intermediate germ insects, only the anterior segments are specified during the blastoderm stage, leaving the posterior segments to be specified later, during embryogenesis, which differs from the segmentation process in Drosophila, a long germ insect. To elucidate the segmentation mechanisms of short and intermediate germ insects, we have investigated the orthologs of the Drosophila segmentation genes in a phylogenetically basal, intermediate germ insect, Gryllus bimaculatus (Gb). Here, we have focused on its hunchback ortholog (Gb'hb), because Drosophila hb functions as a gap gene during anterior segmentation, referred as a canonical function. Gb'hb is expressed in a gap pattern during the early stages of embryogenesis, and later in the posterior growth zone. By means of embryonic and parental RNA interference for Gb'hb, we found the following: (1) Gb'hb regulates Hox gene expression to specify regional identity in the anterior region, as observed in Drosophila and Oncopeltus; (2) Gb'hb controls germband morphogenesis and segmentation of the anterior region, probably through the pair-rule gene, even-skipped at least; (3) Gb'hb may act as a gap gene in a limited region between the posterior of the prothoracic segment and the anterior of the mesothoracic segment; and (4) Gb'hb is involved in the formation of at least seven abdominal segments, probably through its expression in the posterior growth zone, which is not conserved in Drosophila. These findings suggest that Gb'hb functions in a non-canonical manner in segment patterning. A comparison of our results with the results for other derived species revealed that the canonical hb function may have evolved from the non canonical hb functions during evolution. PMID- 15788458 TI - Shh-dependent formation of the ZLI is opposed by signals from the dorsal diencephalon. AB - The zona limitans intrathalamica (ZLI) is located at the border between the prospective ventral thalamus and dorsal thalamus, and functions as a diencephalic signaling center. Little is known about the mechanism controlling ZLI formation. Using a combination of fate-mapping studies and in vitro assays, I show that the differentiation of the ZLI from progenitor cells in the alar plate is initiated by a Shh-dependent signal from the basal plate. The subsequent dorsal progression of ZLI differentiation requires ongoing Shh signaling, and is constrained by inhibitory factors derived from the dorsal diencephalon. These studies demonstrate that self-organizing signals from the basal plate regulate the formation of a potential patterning center in the ZLI in an orthogonal orientation in the alar plate, and thus create the potential for coordinated thalamic patterning in two dimensions. PMID- 15788459 TI - Commissural neuron identity is specified by a homeodomain protein, Mbh1, that is directly downstream of Math1. AB - Proneural basic helix-loop-helix (bHLH) proteins are key regulators of neurogenesis. However, downstream target genes of the bHLH proteins remain poorly defined. Mbh1 confers commissural neuron identity in the spinal cord. Enhancer analysis using transgenic mice revealed that Mbh1 expression required an E-box 3' of the Mbh1 gene. Mbh1 expression was lost in Math1 knockout mice, whereas misexpression of Math1 induced ectopic expression of Mbh1. Moreover, Math1 bound the Mbh1 enhancer containing the E-box in vivo and activated gene expression. Generation of commissural neurons by Math1 was inhibited by a dominant negative form of Mbh1. These findings indicate that Mbh1 is necessary and sufficient for the specification of commissural neurons, as a direct downstream target of Math1. PMID- 15788461 TI - Assessment of the quality of breast cancer care: a single institutional study from Saudi Arabia. AB - OBJECTIVE: To evaluate the quality of operable breast cancer care in a tertiary care institution. DESIGN: A retrospective analysis of all breast cancer patients seen in our institution between 1995 and 2000. Data were abstracted from the charts of these patients. Indicators were based on an international consensus conference and other publications. SETTING: A tertiary care health care institution. MAIN MEASURES: We evaluated the charts and calculated the percentage for which the internationally accepted quality care indicators were followed during the continuum of care. We also reviewed the histopathological reports to evaluate conformation with the accepted indicators. RESULTS: Charts of 75 patients (four exclusions, three metastatic, and one male), diagnosed to have breast cancer during the study period were reviewed. Only 28 (37%) patients had triple assessment before a definitive surgical procedure. Pre-operative staging including a CT and bone scan was performed in 58 (77.3%). Among the 50 patients who had definite surgical intervention, the majority had mastectomy (44/50, 88%) whereas axillary dissection was performed in 46 (46/50, 92%). Estrogen and progesterone receptor status was reported in only four (4/50, 8%) and the exact tumor size in 24 (24/50, 48%) of the histopathological reports. Adjuvant chemotherapy was used in accordance with the international standards but radiotherapy was under-utilized. CONCLUSION: Our study demonstrated that the quality of breast cancer care in this institution was below the accepted international standards. This study may be used to make interventions for improvement of quality in similar institutions all over the kingdom. PMID- 15788460 TI - Six1 and Six4 homeoproteins are required for Pax3 and Mrf expression during myogenesis in the mouse embryo. AB - In mammals, Six5, Six4 and Six1 genes are co-expressed during mouse myogenesis. Six4 and Six5 single knockout (KO) mice have no developmental defects, while Six1 KO mice die at birth and show multiple organ developmental defects. We have generated Six1Six4 double KO mice and show an aggravation of the phenotype previously reported for the single Six1 KO. Six1Six4 double KO mice are characterized by severe craniofacial and rib defects, and general muscle hypoplasia. At the limb bud level, Six1 and Six4 homeogenes control early steps of myogenic cell delamination and migration from the somite through the control of Pax3 gene expression. Impaired in their migratory pathway, cells of the somitic ventrolateral dermomyotome are rerouted, lose their identity and die by apoptosis. At the interlimb level, epaxial Met expression is abolished, while it is preserved in Pax3-deficient embryos. Within the myotome, absence of Six1 and Six4 impairs the expression of the myogenic regulatory factors myogenin and Myod1, and Mrf4 expression becomes undetectable. Myf5 expression is correctly initiated but becomes restricted to the caudal region of each somite. Early syndetomal expression of scleraxis is reduced in the Six1Six4 embryo, while the myotomal expression of Fgfr4 and Fgf8 but not Fgf4 and Fgf6 is maintained. These results highlight the different roles played by Six proteins during skeletal myogenesis. PMID- 15788462 TI - The insufficiency of evidence to establish the business case for quality. AB - PURPOSE: To determine whether a positive financial return on investment for quality-enhancing interventions is more likely for particular health conditions, in specific organizational settings, or with the use of particular interventions. DATA SOURCES: Electronic search of MEDLINE. DATA EXTRACTION: Search keywords included: business case, cost-effectiveness, cost-benefit, return on investment, costs, cost savings, quality, quality improvement, and program evaluation. RESULTS: Only 15 of 1968 articles identified contained sufficient information on both the costs of implementing quality-enhancing interventions and the resultant changes in costs of care or revenues to permit the calculation of a return on investment. CONCLUSIONS: Scant attention is currently paid in the quality-of-care literature to the cost of implementing quality-enhancing interventions. To understand which quality-enhancing interventions are likely to produce positive returns on investments, data collection and analysis must include tracking the investment and operating costs of implementing the intervention as well as the changes in revenues and costs that result from the intervention. PMID- 15788463 TI - The association between quality of care and technical efficiency in long-term care. AB - OBJECTIVE: To analyse the association between quality of care and technical (productive) efficiency in institutional long-term care wards for the elderly. SETTING: One hundred and fourteen public health centre hospitals and residential homes in Finland. STUDY DESIGN: Wards were divided into two categories according to their rank in the quality distribution, considering 41 quality variables separately. The technical efficiency scores of the good- and poor-quality groups were compared using cross-sectional data. METHODS: Data envelopment analysis was used for calculating technical efficiency. The Mann-Whitney test and correlation coefficients were used to explore the association between quality and efficiency. RESULTS: The wards where quality indicators indicated less pro-active (passive) nursing practice and more dependent patients-for instance, in terms of very high prevalence of bedfast residents or very high prevalence of daily physical restraints-performed more efficiently than the comparison group. CONCLUSION: The results suggest that an association may exist between technical efficiency and unwanted dimensions of quality. Hence, the efficiency and quality of care are essential aspects of management and performance measurement in elderly care. PMID- 15788464 TI - Towards quality indicators for assertive outreach programmes for severely impaired substance abusers: concept mapping with Dutch experts. AB - OBJECTIVE: We investigated the concept of 'quality of assertive outreach programmes for severely impaired substance abusers' with the aim of developing a conceptual framework as the basis for an assessment instrument. DESIGN: We held a concept-mapping session with 13 experts in 2003. Fifty measurable elements of quality were mentioned and rated in terms of relative importance on a Likert-type response scale. Subsequently, the experts grouped the statements that were similar in content. The resulting concept map and additional interpretation made up the final quality framework. SETTING/STUDY PARTICIPANTS: Theoretical sampling was used to select Dutch managers, team leaders, and service providers from different assertive outreach delivery systems for substance abusers. Variation in both perspective and region was reflected in the sample. RESULTS: Nine aspects of quality were formulated: preconditions for care, preconditions for service providers' work, relationship to regular care, service providers' activities and goals, service providers' skills, the role of repression, optimal care for the client, goals of assertive outreach, and nuisance reduction to society. Each aspect was presented using a selection of measurable elements. CONCLUSIONS: According to the experts, optimal assertive outreach depends on a broad range of aspects that were later classified in three regions: structure, process, and outcomes. Saturation of the elements has not been proved so far. Nevertheless, it is promising that the framework's regions are supported by theory and that it is largely in accordance with clients' perspectives on assertive community treatment. PMID- 15788465 TI - Differences in quality of care among patients hospitalized with atrial fibrillation as primary or secondary cause for admission. AB - OBJECTIVE: Several factors have been linked to the variation in the quality of care for patients with atrial fibrillation (AF). Whether hospitalization primarily for AF (primary diagnosis of AF) as opposed to another primary diagnosis but having concomitant AF (secondary diagnosis of AF) impacts quality of care for AF is not known. Accordingly, we sought to evaluate the differences in quality of care of Medicare patients admitted with primary diagnosis versus secondary diagnosis of AF. DESIGN AND SETTING: We studied a random sample of Medicare fee-for-service discharges from Michigan's acute care hospitals over a 1 year period with a primary or secondary diagnosis of AF (ICD-9-CM 427.31). Main outcome measure. Warfarin use at the time of discharge. RESULTS: Of 5993 patients in the study, 772 had a primary diagnosis of AF and 5221 had a secondary diagnosis of AF. Patients with a secondary diagnosis of AF were older, more likely to be male, and less likely to be hypertensive. Patients with a secondary diagnosis of AF 'ideal' for anticoagulation (n = 1648) were less likely to receive warfarin compared with 'ideal' patients with primary diagnosis of AF (n = 363) (52.6% versus. 59.8%, P < 0.001). Adherence to test indicators was lower in patients with secondary diagnosis of AF. CONCLUSION: Secondary diagnosis of AF rather than AF as a primary diagnosis appears to account for most Medicare patients with AF admitted to hospitals. Whereas quality of care is lower in patients with secondary diagnosis of AF, opportunity for quality improvement exists for both groups of patients with AF. PMID- 15788466 TI - Variations in the quality of care of patients with acute myocardial infarction among Swiss university hospitals. AB - OBJECTIVE: The objective of our study was to assess hospital variations in the quality of care delivered to acute myocardial infarction (AMI) patients among three Swiss academic medical centres. DESIGN: Cross-sectional study. SETTING: Three Swiss university hospitals. STUDY PARTICIPANTS: We selected 1129 eligible patients discharged from these hospitals from 1 January to 31 December 1999, with a primary or secondary diagnosis code [International Classification of Diseases, 10th revision (ICD-10)] of AMI. We abstracted medical records for information on demographic characteristics, risk factors, symptoms, and findings at admission. We also recorded the main ECG and laboratory findings, as well as hospital and discharge management and treatment. We excluded patients transferred to another hospital and who did not meet the clinical definition of AMI. MAIN OUTCOME MEASURES: Percentage of patients receiving appropriate intervention as defined by six quality of care indicators derived from clinical practical guidelines. RESULTS: Among 577 eligible patients with AMI in this study, the mean (SD) age was 68.2 (13.9), and 65% were male. In the assessment of the quality indicators we excluded patients who were not eligible for the procedure. Among cohorts of 'ideal candidates' for specific interventions, 64% in hospital A and 73% in hospital C had reperfusion within 12 hours either with thrombolytics or percutaneous transluminal coronary angioplasty (P = 0.367). Further, in hospitals A, B, and C, respectively 97, 94, and 84% were prescribed aspirin during the initial hospitalization (P = 0.0002), and respectively 68, 91, and 75% received angiotensin converting enzyme inhibitors at discharge in the case of left ventricular systolic dysfunction (P = 0.003). CONCLUSIONS: Our results showed important hospital-to-hospital variations in the quality of care provided to patients with AMI between these three university hospitals. PMID- 15788468 TI - Type of vascular access and survival among incident hemodialysis patients: the Choices for Healthy Outcomes in Caring for ESRD (CHOICE) Study. AB - Arteriovenous fistulae (AVF) have advantages over arteriovenous grafts (AVG) and central venous catheters (CVC), but whether AVF are associated independently with better survival is unclear. Recent studies showing such a survival benefit did not include early access experience or account for changes in access type over time and did not include data on some important confounders. Reported here are survival rates stratified by the type of access in use up to 3 yr after initiation of hemodialysis among 616 incident patients who were enrolled in the Choices for Healthy Outcomes in Caring for ESRD (CHOICE) Study. A total of 1084 accesses (185 AVF, 296 AVG, 603 CVC) were used for a total of 1381 person-years. At initiation, 409 (66%) patients were using a CVC, 122 (20%) were using an AVG, and 85 (14%) were using an AVF. After 6 mo, 34% were using a CVC, 40% were using an AVG, and 26% were using an AVF. Annual mortality rates were 11.7% for AVF, 14.2% for AVG, and 16.1% for CVC. Adjusted relative hazards (RH) of death compared with AVF were 1.5 (95% confidence interval, 1.0 to 2.2) for CVC and 1.2 (0.8 to 1.8) for AVG. The increased hazards associated with CVC, as compared with AVF, were stronger in men (n = 334; RH = 2.0; P = 0.01) than women (n = 282; RH = 1.0 for CVC; P = 0.92). These results strongly support existing clinical practice guidelines and suggest that the use of venous catheters should be minimized to reduce the frequency of access complications and to improve patient survival, especially among male hemodialysis patients. PMID- 15788469 TI - Structured conversion from thrice weekly to weekly erythropoietic regimens using a computerized decision-support system: a randomized clinical study. AB - In view of the recent interest in weekly erythropoietic regimens and the lack of studies directly comparing the available agents, the clinical effectiveness of darbepoetin-alpha (DA) and epoetin-beta (EB), when administered via the subcutaneous route on a weekly basis, after conversion from thrice-weekly subcutaneous EB, was studied. In this 9-mo, single-center, randomized study of an unselected hemodialysis population, anemia was managed with a computerized decision-support system. Per-protocol analysis of the 81 patients in each arm who completed the study showed similar hemoglobin outcomes between treatment arms, both at randomization and at the end of the study. After conversion from thrice weekly EB to DA (at a ratio of 200 IU:1 microg, at which products are cost neutral in the European Union), a significant fall in dose from a mean of 0.59 microg/kg per wk after randomization to 0.46 microg/kg per wk in the last month (P = 0.002) was observed; in the comparator arm, the reduction in frequency of administration of EB was associated with a significant dose increase from a mean of 107.5 to 133.2 IU/kg per wk (P = 0.002) during the same period. At hemoglobin stability, mean EB dose was found to be 44% higher than DA dose (when multiplied by 200). Similar significant dose differences were apparent in a modified intention-to-treat analysis. The study demonstrated that, under a decision support system, both products were capable of adequately maintaining hemoglobin outcome when administered on a weekly basis but with significant dose differences at 9 mo. PMID- 15788470 TI - Incidence of atherosclerosis by race in the dialysis morbidity and mortality study: a sample of the US ESRD population. AB - White individuals who are on dialysis experience much higher overall and cardiovascular mortality rates than black individuals despite a more favorable risk factor profile, but the incidence of nonfatal cardiovascular disease (CVD) to this racial disparity has not been well studied. A longitudinal study of 16,103 people who had ESRD and were enrolled in the United Renal Data System from 1993 to 1996 was conducted. The incidence of new and recurrent atherosclerotic CVD (ASCVD) events was determined using Medicare claims for hospitalizations and mortality among blacks and whites, stratified by ASCVD at baseline. ASCVD was defined as coronary heart disease, peripheral vascular disease, and cerebrovascular disease. Incidence of new ASCVD in people without ASCVD at baseline was 146.9 per 1000 person-years in whites and 118.7 per 1000 person years in blacks. Incidence of recurrent ASCVD was 404.1 per 1000 person-years in whites and 317.5 per 1000 person-years in blacks. Whites were 1.35 (95% confidence interval, 1.18 to 1.55) times more likely to develop incident ASCVD compared with blacks and 1.25 (95% confidence interval, 1.14 to 1.36) times more likely to develop recurrent disease after adjusting for traditional CVD and dialysis-related risk factors. Excess risk for recurrent ASCVD in whites compared with blacks was consistently present no matter the duration of dialysis: Hazard ratio 1.42 for 0 to 6 mo and 1.40 for 6 to 12 mo. Whites who are treated with dialysis have a higher incidence of ASCVD than blacks who are on dialysis, both new and recurrent. Although differences in survival before the initiation of dialysis may contribute to the observed difference in ASCVD risk, it is not explained by baseline traditional and dialysis-related ASCVD risk factors. PMID- 15788472 TI - Novel serpinopathy in rat kidney and pancreas induced by overexpression of megsin. AB - The intracellular polymerization of abnormal serine protease inhibitors (serpins) results in liver or neuronal cell abnormalities recently identified as "serpinopathies." It was demonstrated in transgenic rats that overexpression of megsin, a recently discovered serpin located in the kidney, produces renal and pancreatic lesions characteristic of serpinopathies. Megsin expression is elevated in a variety of organs, including kidney and pancreas. Periodic acid Schiff-positive, diastase-resistant intracellular inclusions develop only in the kidney and the pancreas. They correspond to electron-dense deposits, shown to contain megsin by immunohistochemistry and immunoelectron microscopy. In the kidney, inclusions are located mainly in the endoplasmic reticulum of glomerular epithelial, distal, and collecting duct cells, and are associated with massive proteinuria and an impaired renal function. In the pancreas, similar inclusions are found in the exocrine and Langerhans islet cells, where islet beta cells are reduced as a result of apoptosis. They are associated with diabetes with low insulin levels. The animals have an impaired growth and die within 10 wk. Rats that overexpress a mutant megsin, characterized by a deficient conformational transition activity, do not develop the serpinopathy, suggesting that some conformational flexibility of the serpin is required for the development of serpinopathy. This model of serpinopathy is the first to involve the kidney and the pancreas. PMID- 15788471 TI - Graded effects of proteinuria on HDL structure in nephrotic rats. AB - Nephrotic syndrome is characterized by increased triglycerides resulting from decreased clearance of VLDL and chylomicrons. These triglyceride-rich lipoproteins are structurally altered by interaction with HDL derived from animals with proteinuria and not as a consequence of hypoalbuminemia. HDL isolated from rats with massive proteinuria is depleted in apolipoprotein E (apoE). It is unknown at what threshold of urinary albumin loss HDL structure is altered, and it is unknown what effects proteinuria has on apolipoproteins other than apoE. Two models of albuminuria were used in Sprague-Dawley rats: Adriamycin and passive Heymann nephritis (HN). The adriamycin group was divided into minimal albumin excretion (MAE) and intermediate albumin excretion (MAE, 1 to 40; intermediate albumin excretion, 60 to 210 mg/d per 100 g body wt). Urinary albumin excretion exceeded 300 mg/d per 100 g body wt in the HN rats. HDL apolipoprotein composition was analyzed with SDS-PAGE densitometry and liquid chromatography-time of flight mass spectrometer mass spectrometry. HDL apoA-IV content relative to apoA-I was reduced at all levels of albuminuria (P < 0.0001). ApoE was not reduced in MAE but was significantly reduced in IAE (72%; P < 0.001). By contrast, apoA-II and apoC-III were each significantly increased with increasing UAE. ApoA-IV and apoE were decreased to approximately 10% of control in HDL isolated from rats with HN, whereas apoA-II, apoC-II, and apoC-III were each significantly increased relative to apoA-I. HDL is structurally altered by levels of albuminuria that are insufficient to change serum albumin levels and is progressively altered as albuminuria increases. PMID- 15788473 TI - Localization and functional characterization of glycosaminoglycan domains in the normal human kidney as revealed by phage display-derived single chain antibodies. AB - Glycosaminoglycans (GAG) play an important role in renal homeostasis. They are strongly negatively charged polysaccharides that bind and modulate a myriad of proteins, including growth factors, cytokines, and enzymes. With the aid of specific phage display-derived antibodies, the distribution of heparan sulfate (HS) and chondroitin sulfate (CS) domains in the normal human kidney was studied. HS domains were specifically located in basement membranes and/or surfaces of renal cells and displayed a characteristic distribution over the nephron. A characteristic location in specific parts of the tubular system was also observed. CS showed mainly an interstitial location. Immunoelectron microscopy indicated specific ultrastructural location of domains. Only partial overlap with any of seven different proteoglycan core proteins was observed. Two HS domains, one highly sulfated (defined by antibody HS4C3) and one low sulfated (defined by antibody RB4Ea12), were studied for their cell biologic relevance with respect to the proliferative effect of FGF-2 on human mesangial cells in vitro. Fibroblast growth factor 2 (FGF-2) binding was HS dependent. Addition of purified HS4C3 antibody but not of the RB4Ea12 antibody counteracted the binding and the proliferative effect of FGF-2, indicating that the HS4C3 domain is involved in FGF-2 handling by mesangial cells. In conclusion, specific GAG domains are differentially distributed in the normal human kidney and are likely involved in binding of effector molecules such as FGF-2. The availability of tools to identify and study relevant GAG structures allows the development of glycomimetica to halt, for instance, mesangial proliferation and matrix production as seen in diabetic nephropathy. PMID- 15788475 TI - Randomized, controlled trial of topical exit-site application of honey (Medihoney) versus mupirocin for the prevention of catheter-associated infections in hemodialysis patients. AB - The clinical usefulness of hemodialysis catheters is limited by increased infectious morbidity and mortality. Topical antiseptic agents, such as mupirocin, are effective at reducing this risk but have been reported to select for antibiotic-resistant strains. The aim of the present study was to determine the efficacy and the safety of exit-site application of a standardized antibacterial honey versus mupirocin in preventing catheter-associated infections. A randomized, controlled trial was performed comparing the effect of thrice-weekly exit-site application of Medihoney versus mupirocin on infection rates in patients who were receiving hemodialysis via tunneled, cuffed central venous catheters. A total of 101 patients were enrolled. The incidences of catheter associated bacteremias in honey-treated (n = 51) and mupirocin-treated (n = 50) patients were comparable (0.97 versus 0.85 episodes per 1000 catheter-days, respectively; NS). On Cox proportional hazards model analysis, the use of honey was not significantly associated with bacteremia-free survival (unadjusted hazard ratio, 0.94; 95% confidence interval, 0.27 to 3.24; P = 0.92). No exit-site infections occurred. During the study period, 2% of staphylococcal isolates within the hospital were mupirocin resistant. Thrice-weekly application of standardized antibacterial honey to hemodialysis catheter exit sites was safe, cheap, and effective and resulted in a comparable rate of catheter-associated infection to that obtained with mupirocin (although the study was not adequately powered to assess therapeutic equivalence). The effectiveness of honey against antibiotic-resistant microorganisms and its low likelihood of selecting for further resistant strains suggest that this agent may represent a satisfactory alternative means of chemoprophylaxis in patients with central venous catheters. PMID- 15788474 TI - Signaling mechanism of TGF-beta1 in prevention of renal inflammation: role of Smad7. AB - TGF-beta has been shown to play a critical role in anti-inflammation; however, the signaling mechanisms of TGF-beta in anti-inflammatory response remains largely unclear. This study reported that mice that overexpress latent TGF-beta1 on skin are protected against renal inflammation in a model of obstructive kidney disease and investigated the signaling mechanism of TGF-beta1 in inhibition of renal inflammation in vivo and in vitro. Seven days after urinary obstruction, wild-type mice developed severe renal inflammation, including massive T cell and macrophage infiltration and marked upregulation of IL-1beta, TNF-alpha, and intercellular adhesion molecule-1 (all P < 0.001). Surprising, renal inflammation was prevented in transgenic mice. This was associated with an increase in latent TGF-beta1 in circulation (a 10-fold increase) and renal tissues (a 2.5-fold increase). Further studies showed that inhibition of renal inflammation in TGF beta1 transgenic mice was also associated with a marked upregulation of renal Smad7 and IkappaBalpha and a suppression of NF-kappaB activation in the diseased kidney (all P < 0.01). These in vivo findings suggested the importance of TGF beta-NF-kappaB cross-talk signaling pathway in regulating renal inflammation. This was tested in vitro in a doxycycline-regulated Smad7-expressing renal tubular cell line. Overexpression of Smad7 was able to upregulate IkappaBalpha directly in a time- and dose-dependent manner, thereby inhibiting NF-kappaB activation and NF-kappaB-driven inflammatory response. In conclusion, latent TGF beta may have protective roles in renal inflammation. Smad7-mediated inhibition of NF-kappaB activation via the induction of IkBalpha may be the central mechanism by which latent TGF-beta prevents renal inflammation. PMID- 15788476 TI - Regulation of renal calcium receptor gene expression by 1,25-dihydroxyvitamin D3 in genetic hypercalciuric stone-forming rats. AB - Hypercalciuria in inbred genetic hypercalciuric stone-forming (GHS) rats is due, in part, to a decrease in renal tubule Ca reabsorption. Activation of the renal Ca receptor (CaR) may decrease renal tubule Ca reabsorption and cause hypercalciuria through suppression of Ca-sensitive potassium channel activity. Because the rat renal CaR gene is regulated by extracellular calcium and 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] and GHS rats have increased renal vitamin D receptor content, the current study was undertaken to determine the level of CaR gene expression in GHS rat kidney and whether CaR gene expression is regulated by 1,25(OH)2D3. Male GHS and normal control (NC) rats were fed a Ca-sufficient diet (0.6% Ca). Western blotting revealed a four-fold increase in CaR protein in GHS rat renal tissue, and 1,25(OH)2D3 administration increased renal CaR in both GHS and NC rats. Northern blot analysis of extracts of renal cortical tissue from GHS and NC rats revealed a major 7-kb transcript of CaR and a more modest 4-kb transcript, both of which were readily detectable. Both Northern blotting and real-time reverse transcription-PCR revealed increased basal CaR mRNA expression levels in GHS rat kidney. 1,25(OH)2D3 administration increased renal CaR mRNA levels 2.0- and 3.3-fold in GHS and NC rats, respectively. Despite the greater incremental increase by 1,25(OH)2D3 in NC rats, CaR mRNA levels remained higher in GHS rat kidney, and the elevation was more sustained. 1,25(OH)2D3 increased CaR mRNA through both elevated CaR gene expression and prolonged tissue half life. These results demonstrate that GHS rats have high levels of CaR gene expression and CaR protein that may contribute to the hypercalciuria and calcium nephrolithiasis. PMID- 15788477 TI - Role of mammalian target of rapamycin signaling in compensatory renal hypertrophy. AB - Loss of functioning nephrons stimulates the growth of residual kidney tissue to augment work capacity and maintain normal renal function. This growth largely occurs by hypertrophy rather than from hyperplasia of the remaining nephrons. The signaling mechanisms that increase RNA and protein synthesis during compensatory renal hypertrophy are unknown. This study found that the remaining kidney hypertrophied 42% by 16 d after unilateral nephrectomy (UNX) in DBA/2 mice. Immunoblotting analysis revealed increased phosphorylation of the 40S ribosomal protein S6 (rpS6) and the eukaryotic translation initiation factor (eIF) 4E binding protein 1 (4E-BP1), the two downstream effectors of the mammalian target of rapamycin (mTOR). The highly specific mTOR inhibitor rapamycin blocked UNX increased phosphorylation of both rpS6 and 4E-BP1. UNX increased the content of not only 40S and 60S ribosomal subunits but also 80S monosomes and polysomes in the remaining kidney. Administration of rapamycin decreased UNX-induced polysome formation and shifted the polysome profile in the direction of monosomes and ribosomal subunits. Pretreatment of the mice with rapamycin inhibited UNX-induced hypertrophy. These studies demonstrate that activation of the mTOR signaling pathway in the remaining kidney after UNX plays an essential role in modulating RNA and protein synthesis during development of compensatory renal hypertrophy. PMID- 15788478 TI - Detection of renal function decline in patients with diabetes and normal or elevated GFR by serial measurements of serum cystatin C concentration: results of a 4-year follow-up study. AB - Research on early renal function decline in diabetes is hampered by lack of simple tools for detecting trends (particularly systematic decreases) in renal function over time when GFR is normal or elevated. This study sought to assess how well serum cystatin C meets that need. Thirty participants with type 2 diabetes in the Diabetic Renal Disease Study met these three eligibility criteria: GFR >20 ml/min per 1.73 m2 at baseline (based on cold iothalamate clearance), 4 yr of follow-up, and yearly measurements of iothalamate clearance and serum cystatin C. With the use of linear regression, each individual's trend in renal function over time, expressed as annual percentage change in iothalamate clearance, was determined. Serum cystatin C in mg/L was transformed to its reciprocal (100/cystatin C), and linear regression was used to determine each individual's trend over time, expressed as annual percentage change. In paired comparisons of 100/cystatin C with iothalamate clearance at each examination, the two measures were numerically similar. More important, the trends in 100/cystatin C and iothalamate clearance were strongly correlated (Spearman r = 0.77). All 20 participants with negative trends in iothalamate clearance (declining renal function) also had negative trends for 100/cystatin C. Results were discordant for only three participants. In contrast, the trends for three commonly used creatinine-based estimates of GFR compared poorly with trends in iothalamate clearance (Spearman r < 0.35). Serial measures of serum cystatin C accurately detect trends in renal function in patients with normal or elevated GFR and provide means for studying early renal function decline in diabetes. PMID- 15788479 TI - CD4+CD25+ regulatory T cells inhibit experimental anti-glomerular basement membrane glomerulonephritis in mice. AB - CD4+CD25+ regulatory T cells (Treg) are of critical importance for the maintenance of tolerance. The kidney is frequently involved in autoimmune diseases, such as lupus erythematosus or glomerulonephritis (GN). Therefore, the therapeutic efficacy of Treg in a T cell-dependent murine model of experimental anti-glomerular basement membrane (anti-GBM) GN was tested. Transfer of 1 x 10(6) CD4+CD25+ T cells (day -1) into mice that were previously immunized with rabbit IgG (day -3) and subsequently received an injection of anti-GBM rabbit serum (day 0) significantly attenuated the development of proteinuria when compared with animals that received an injection of 1 x 10(6) CD4+CD25- T cells (control group). Treg injection induced a dramatic decrease of glomerular damage as well as a marked decrease of CD4+ T cell, CD8+ T cell, and macrophage infiltration. Of note, deposition of immune complexes was not prevented by Treg, showing that Treg rather inhibited cell-mediated organ damage than priming of the humoral immune response. Accordingly, a significant reduction of IFN-gamma, TNF-alpha, and TGF beta1 mRNA in kidneys from animals that received Treg injection was observed. Tracking of enhanced green fluorescence protein-transgenic Treg revealed a predominant migration to secondary lymphoid organs with a significant increase of regulatory T cells (CD4+CD25+CD69-CD45RB(low)) in the lymph nodes. In contrast, enhanced green fluorescence protein-and FoxP3-positive cells by reverse transcription-PCR and CD4+CD25+CD69-CD45RB(low) T cells by flow cytometry in the kidney of nephritic animals were not detected. This report provides first evidence that Treg are potent suppressors of anti-GBM GN. Treg therefore might be of therapeutic value for the treatment of severe GN in humans. PMID- 15788481 TI - Cytoskeletal interactions regulate inducible L-selectin clustering. AB - L-selectin (CD62L) amplifies neutrophil capture within the microvasculature at sites of inflammation. Activation by G protein-coupled stimuli or through ligation of L-selectin promotes clustering of L-selectin and serves to increase its adhesiveness, signaling, and colocalization with beta(2)-integrins. Currently, little is known about the molecular process regulating the lateral mobility of L-selectin. On neutrophil stimulation, a progressive change takes place in the organization of its plasma membrane, resulting in membrane domains that are characteristically enriched in glycosyl phosphatidylinositol (GPI) anchored proteins and exclude the transmembrane protein CD45. Clustering of L selectin, facilitated by E-selectin engagement or antibody cross-linking, resulted in its colocalization with GPI-anchored CD55, but not with CD45 or CD11c. Disrupting microfilaments in neutrophils or removing a conserved cationic motif in the cytoplasmic domain of L-selectin increased its mobility and membrane domain localization in the plasma membrane. In addition, the conserved element was critical for L-selectin-dependent tethering under shear flow. Our data indicate that L-selectin's lateral mobility is regulated by interactions with the actin cytoskeleton that in turn fortifies leukocyte tethering. We hypothesize that both membrane mobility and stabilization augment L-selectin's effector functions and are regulated by dynamic associations with membrane domains and the actin cytoskeleton. PMID- 15788482 TI - Ezrin oligomers are the membrane-bound dormant form in gastric parietal cells. AB - Ezrin is a member of ezrin, radixin, moesin (ERM) protein family that links F actin to membranes. The NH(2)- and COOH-terminal association domains of ERM proteins, known respectively as N-ERMAD and C-ERMAD, participate in interactions with membrane proteins and F-actin, and intramolecular and intermolecular interactions within and among ERM proteins. In gastric parietal cells, ezrin is heavily represented on the apical membrane and is associated with cell activation. Ezrin-ezrin interactions are presumably involved in functional regulation of ezrin and thus became a subject of our study. Fluorescence resonance energy transfer (FRET) was examined with cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP)-tagged ezrin incorporated into HeLa cells and primary cultures of parietal cells. Constructs included YFP at the NH(2) terminus of ezrin (YFP-Ez), CFP at the COOH terminus of ezrin (Ez-CFP), and double-labeled ezrin (N-YFP-ezrin-CFP-C). FRET was probed using fluorescence microscopy and spectrofluorometry. Evidence of ezrin oligomer formation was found using FRET in cells coexpressing Ez-CFP and YFP-Ez and by performing coimmunoprecipitation of endogenous ezrin with fluorescent protein-tagged ezrin. Thus intermolecular NH(2)- and COOH-terminal association domain (N-C) binding in vivo is consistent with the findings of earlier in vitro studies. After the ezrin oligomers were separated from monomers, FRET was observed in both forms, indicating intramolecular and intermolecular N-C binding. When the distribution of native ezrin as oligomers vs. monomers was examined in resting and maximally stimulated parietal cells, a shift of ezrin oligomers to the monomeric form was correlated with stimulation, suggesting that ezrin oligomers are the membrane bound dormant form in gastric parietal cells. PMID- 15788483 TI - Parathyroid hormone treatment induces dissociation of type IIa Na+-P(i) cotransporter-Na+/H+ exchanger regulatory factor-1 complexes. AB - The type IIa Na+-P(i) cotransporter (NaP(i)-IIa) and the Na+/H+ exchanger regulatory factor-1 (NHERF1) colocalize in the apical membrane of proximal tubular cells. Both proteins interact in vitro. Herein the interaction between NaP(i)-IIa and NHERF1 is further documented on the basis of coimmunoprecipitation and co-pull-down assays. NaP(i)-IIa is endocytosed and degraded in lysosomes upon parathyroid hormone (PTH) treatment. To investigate the effect of PTH on the NaP(i)-IIa-NHERF1 association, we first compared the localization of both proteins after PTH treatment. In mouse proximal tubules and OK cells, NaP(i)-IIa was removed from the apical membrane after hormonal treatment; however, NHERF1 remained at the membrane. Moreover, PTH treatment led to degradation of NaP(i) IIa without changes in the amount of NHERF1. The effect of PTH on the NaP(i)-IIa NHERF1 interaction was further studied using coimmunoprecipitation. PTH treatment reduced the amount of NaP(i)-IIa coimmunoprecipitated with NHERF antibodies. PTH induced internalization of NaP(i)-IIa requires PKA and PKC; therefore, we next analyzed whether PTH induces changes in the phosphorylation state of either partner. NHERF1 was constitutively phosphorylated. Moreover, in mouse kidney slices, PTH induced an increase in NHERF1 phosphorylation; independent activation of PKA or PKC also resulted in increased phosphorylation of NHERF1 in kidney slices. However, NaP(i)-IIa was not phosphorylated either basally or after exposure to PTH. Our study supports an interaction between NHERF1 and NaP(i)-IIa on the basis of their brush-border membrane colocalization and in vitro coimmunoprecipitation/co-pull-down assays. Furthermore, PTH weakens this interaction as evidenced by different in situ and in vivo behavior. The PTH effect takes place in the presence of increased phosphorylation of NHERF1. PMID- 15788484 TI - Reactive oxygen species formation in the transition to hypoxia in skeletal muscle. AB - Many tissues produce reactive oxygen species (ROS) during reoxygenation after hypoxia or ischemia; however, whether ROS are formed during hypoxia is controversial. We tested the hypothesis that ROS are generated in skeletal muscle during exposure to acute hypoxia before reoxygenation. Isolated rat diaphragm strips were loaded with dihydrofluorescein-DA (Hfluor-DA), a probe that is oxidized to fluorescein (Fluor) by intracellular ROS. Changes in fluorescence due to Fluor, NADH, and FAD were measured using a tissue fluorometer. The system had a detection limit of 1 microM H2O2 applied to the muscle superfusate. When the superfusion buffer was changed rapidly from 95% O2 to 0%, 5%, 21%, or 40% O2, transient elevations in Fluor were observed that were proportional to the rise in NADH fluorescence and inversely proportional to the level of O2 exposure. This signal could be inhibited completely with 40 microM ebselen, a glutathione peroxidase mimic. After brief hypoxia exposure (10 min) or exposure to brief periods of H2O2, the fluorescence signal returned to baseline. Furthermore, tissues loaded with the oxidized form of the probe (Fluor-DA) showed a similar pattern of response that could be inhibited with ebselen. These results suggest that Fluor exists in a partially reversible redox state within the tissue. When Hfluor-loaded tissues were contracted with low-frequency twitches, Fluor emission and NADH emission were significantly elevated in a way that resembled the hypoxia induced signal. We conclude that in the transition to low intracellular P(O2), a burst of intracellular ROS is formed that may have functional implications regarding skeletal muscle O2-sensing systems and responses to acute metabolic stress. PMID- 15788485 TI - Etk/Bmx mediates expression of stress-induced adaptive genes VEGF, PAI-1, and iNOS via multiple signaling cascades in different cell systems. AB - We recently showed that Etk/Bmx, a member of the Tec family of nonreceptor protein tyrosine kinases, promotes tight junction formation during chronic hypoxic exposure and augments normoxic VEGF expression via a feedforward mechanism. Here we further characterized Etk's role in potentiating hypoxia induced gene expression in salivary epithelial Pa-4 cells. Using transient transfection in conditionally activated Etk (DeltaEtk:ER) cells, we demonstrated that Etk enhances hypoxia-response element-dependent reporter activation in normoxia and hypoxia. This Etk-driven reporter activation is ameliorated by treatment with wortmannin or LFM-A13. Using lentivirus-mediated gene delivery and small interfering RNA, we provided direct evidence that hypoxia leads to transient Etk and Akt activation and hypoxia-mediated Akt activation is Etk dependent. Northern blot analyses confirmed that Etk activation led to induction of steady-state mRNA levels of endogenous VEGF and plasminogen activator inhibitor (PAI)-1, a hallmark of hypoxia-mediated gene regulation. We also demonstrated that Etk utilizes a phosphatidylinositol 3-kinase/Akt pathway to promote reporter activation driven by NF-kappaB, another oxygen-sensitive transcription factor, and to augment cytokine-induced inducible nitric oxide synthase expression in endothelial cells. To establish the clinical relevance of Etk-induced, hypoxia-mediated gene regulation, we examined Etk expression in keloid, which has elevated VEGF and PAI-1. We found that Etk is overexpressed in keloid (but not normal skin) tissues. The differential steady-state Etk protein levels were further confirmed in primary fibroblast cultures derived from these tissues, suggesting an Etk role in tissue fibrosis. Our results provide further understanding of Etk function within multiple signaling cascades to govern adaptive cytoprotection against extracellular stress in different cell systems, salivary epithelial cells, brain endothelial cells, and dermal fibroblasts. PMID- 15788486 TI - Lipopolysaccharide- and gram-positive bacteria-induced cellular inflammatory responses: role of heterotrimeric Galpha(i) proteins. AB - Heterotrimeric G(i) proteins may play a role in lipopolysaccharide (LPS) activated signaling through Toll-like receptor 4 (TLR4), leading to inflammatory mediator production. Although LPS is a TLR4 ligand, the gram-positive bacterium Staphylococcus aureus (SA) is a TLR2 ligand, and group B streptococci (GBS) are neither TLR2 nor TLR4 ligands but are MyD88 dependent. We hypothesized that genetic deletion of G(i) proteins would alter mediator production induced by LPS and gram-positive bacterial stimulation. We examined genetic deletion of Galpha(i2) or Galpha(i1/3) protein in Galpha(i2)-knockout (Galpha(i2)-/-) or Galpha(i1/3)-knockout (Galpha(i1/3)-/-) mice. LPS-, heat-killed SA-, or GBS induced mediator production in splenocytes or peritoneal macrophages (MPhi) was investigated. There were significant increases in LPS-, SA-, and GBS-induced production of TNF-alpha and IFN-gamma in splenocytes from Galpha(i2)-/- mice compared with wild-type (WT) mice. Also, LPS-induced TNF-alpha was increased in splenocytes from Galpha(i1/3)-/- mice. In contrast to splenocytes, LPS-, SA-, and GBS-induced TNF-alpha, IL-10, and thromboxane B(2) (TxB(2)) production was decreased in MPhi harvested from Galpha(i2)-/- mice. Also, LPS-induced production of IL-10 and TxB(2) was decreased in MPhi from Galpha(i1/3)-/- mice. In subsequent in vivo studies, TNF-alpha levels after LPS challenge were significantly greater in Galpha(i2)-/- mice than in WT mice. Also, myeloperoxidase activity, a marker of tissue neutrophil infiltration, was significantly increased in the gut and lung of LPS-treated Galpha(i2)-/- mice compared with WT mice. These data suggest that G(i) proteins differentially regulate murine TLR-mediated inflammatory cytokine production in a cell-specific manner in response to both LPS and gram-positive microbial stimuli. PMID- 15788487 TI - Bisphosphonates stimulate an endogenous nonselective cation channel in Xenopus oocytes: potential mechanism of action. AB - The mechanisms of action of bisphosphonates (BPs) have been poorly determined. Besides their actions on osteoclasts, these agents exhibit gastrointestinal complications. They have also recently been described as affecting various preparations that express an epithelial Na(+) channel (ENaC). To understand the effects of BP on ion channels and the ENaC in particular, we used the Xenopus oocyte expression system. Alendronate, and similarly risedronate, two aminobisphosphonates, caused a large stimulation of an endogenous nonselective cation conductance (NSCC). This stimulation averaged 63 +/- 12 muS (n = 18) 60 min after the addition of 2 mM alendronate. The effects on the endogenous NSCC were blocked by extracellular acidification to pH 6.4. On the other hand, alendronate caused a small inhibition of ENaC conductance at pH 7.4 and 6.4, but the effects at pH 6.4 were more readily observed in the absence of changes of the endogenous conductance. The effects on membrane capacitance were also markedly different, with a clear decrease at pH 6.4 and no consistent changes at pH 7.4. The effects on the endogenous channel were further augmented by genistein and were inhibited by a tyrosine phosphatase inhibitor, indicating the involvement of the tyrosine kinase pathway. Stimulation of NSCC with BP is expected to cause membrane depolarization and may explain, in part, its mechanisms of action in inhibiting osteoclasts. PMID- 15788488 TI - Expression and functional properties of four slow skeletal troponin T isoforms in rat muscles. AB - We investigated the expression and functional properties of slow skeletal troponin T (sTnT) isoforms in rat skeletal muscles. Four sTnT cDNAs were cloned from the slow soleus muscle. Three isoforms were found to be similar to sTnT1, sTnT2, and sTnT3 isoforms described in mouse muscles. A new rat isoform, with a molecular weight slightly higher than that of sTnT3, was discovered. This fourth isoform had never been detected previously in any skeletal muscle and was therefore called sTnTx. From both expression pattern and functional measurements, it appears that sTnT isoforms can be separated into two classes, high-molecular weight (sTnT1, sTnT2) and low-molecular-weight (sTnTx, sTnT3) isoforms. By comparison to the apparent migration pattern of the four recombinant sTnT isoforms, the newly described low-molecular-weight sTnTx isoform appeared predominantly and typically expressed in fast skeletal muscles, whereas the higher-molecular-weight isoforms were more abundant in slow soleus muscle. The relative proportion of the sTnT isoforms in the soleus was not modified after exposure to hindlimb unloading (HU), known to induce a functional atrophy and a slow-to-fast isoform transition of several myofibrillar proteins. Functional data gathered from replacement of endogenous troponin complexes in skinned muscle fibers showed that the sTnT isoforms modified the Ca(2+) activation characteristics of single skeletal muscle fibers, with sTnT2 and sTnT1 conferring a similar increase in Ca(2+) affinity higher than that caused by low-molecular weight isoforms sTnTx and sTnT3. Thus we show for the first time the presence of sTnT in fast muscle fibers, and our data show that the changes in neuromuscular activity on HU are insufficient to alter the sTnT expression pattern. PMID- 15788489 TI - Beta-adrenergic- and muscarinic receptor-induced changes in cAMP activity in adult cardiac myocytes detected with FRET-based biosensor. AB - beta-Adrenergic receptor activation regulates cardiac myocyte function through the stimulation of cAMP production and subsequent activation of protein kinase A (PKA). Furthermore, muscarinic receptor activation inhibits as well as facilitates these cAMP-dependent effects. However, it has not always been possible to correlate the muscarinic responses with the direct measurement of changes in cellular cAMP activity. Genetically encoded biosensors have recently been developed, making it possible to monitor real-time changes in cAMP and PKA activity at the single cell level. One such biosensor consists of the regulatory and catalytic subunits of PKA labeled with cyan and yellow fluorescent proteins, respectively. Changes in cAMP activity affecting the association of these labeled PKA subunits can be detected as changes in fluorescence resonance energy transfer. In the present study, an adenovirus-based approach was developed to express this recombinant protein complex in adult cardiac myocytes and use it to monitor changes in cAMP activity produced by beta-adrenergic and muscarinic receptor activation. The biosensor expressed with the use of this system is able to detect changes in cAMP activity produced by physiologically relevant levels of beta-adrenergic receptor activation without disrupting normal functional responses. It was also possible to directly demonstrate the complex temporal pattern of inhibitory and stimulatory changes in cAMP activity produced by muscarinic receptor activation in these cells. The adenovirus-based approach we have developed should facilitate the use of this biosensor in studying cAMP and PKA-dependent signaling mechanisms in a wide variety of cell types. PMID- 15788490 TI - Kinetic characterization of tetrapropylammonium inhibition reveals how ATP and Pi alter access to the Na+-K+-ATPase transport site. AB - Current models of the Na(+)-K(+)-ATPase reaction cycle have ATP binding with low affinity to the K(+)-occluded form and accelerating K(+) deocclusion, presumably by opening the inside gate. Implicit in this situation is that ATP binds after closing the extracellular gate and thus predicts that ATP binding and extracellular cation binding to be mutually exclusive. We tested this hypothesis. Accordingly, we needed a cation that binds outside and not inside, and we determined that tetrapropylammonium (TPA) behaves as such. TPA competed with K(+) (and not Na(+)) for ATPase, TPA was unable to prevent phosphoenzyme (EP) formation even at low Na(+), and TPA decreased the rate of EP hydrolysis in a K(+)-competitive manner. Having established that TPA binding is a measurement of extracellular access, we next determined that TPA and inorganic phosphate (P(i)) were not mutually exclusive inhibitors of para-nitrophenylphosphatase (pNPPase) activity, implying that when P(i) is bound, the transport site has extracellular access. Surprisingly, we found that ATP and TPA also were not mutually exclusive inhibitors of pNPPase activity, implying that when the cation transport site has extracellular access, ATP can still bind. This is consistent with a model in which ATP speeds up the conformational changes that lead to intracellular or extracellular access, but that ATP binding is not, by itself, the trigger that causes opening of the cation site to the cytoplasm. PMID- 15788491 TI - Images in clinical medicine. Severe fecal impaction. PMID- 15788492 TI - The trouble with uncompensated hospital care. PMID- 15788493 TI - Safety in numbers--monitoring risk in approved drugs. PMID- 15788494 TI - Herbal medicine in Europe--relaxing regulatory standards. PMID- 15788495 TI - Hospitalization for mental illness among parents after the death of a child. AB - BACKGROUND: The loss of a child is considered one of the most stressful events in the life of a parent. We hypothesized that parental bereavement increases the risk of hospital admission for a psychiatric disorder, especially for affective disorders. METHODS: We studied a cohort of 1,082,503 persons identified from national registers in Denmark who were born between 1952 and 1999 and had at least one child under 18 years of age during the follow-up period, from 1970 to 1999. Parents who lost a child during follow-up were categorized as "bereaved" from the date of death of the child. RESULTS: As compared with parents who did not lose a child, parents who lost a child had an overall relative risk of a first psychiatric hospitalization for any disorder of 1.67 (95 percent confidence interval, 1.53 to 1.83). Bereaved mothers had a higher relative risk of being hospitalized for any psychiatric disorder than bereaved fathers (relative risks, 1.78 [95 percent confidence interval, 1.60 to 1.98] and 1.38 [95 percent confidence interval, 1.17 to 1.63], respectively; P value for interaction, 0.01). The relative risks of hospitalization specifically for affective disorders were 1.91 (95 percent confidence interval, 1.59 to 2.30) and 1.61 (95 percent confidence interval, 1.15 to 2.27) for bereaved mothers and fathers, respectively. Among mothers, the relative risk of being hospitalized for any psychiatric disorder was highest during the first year after the death of the child but remained significantly elevated five years or more after the death. CONCLUSIONS: The risk of psychiatric hospitalization was increased among parents, especially mothers, who lost a child. PMID- 15788497 TI - Day-night pattern of sudden death in obstructive sleep apnea. AB - BACKGROUND: The risk of sudden death from cardiac causes in the general population peaks from 6 a.m. to noon and has a nadir from midnight to 6 a.m. Obstructive sleep apnea is highly prevalent and associated with neurohormonal and electrophysiological abnormalities that may increase the risk of sudden death from cardiac causes, especially during sleep. METHODS: We reviewed polysomnograms and the death certificates of 112 Minnesota residents who had undergone polysomnography and had died suddenly from cardiac causes between July 1987 and July 2003. For four intervals of the day, we compared the rates of sudden death from cardiac causes among people with obstructive sleep apnea and the following: the rates among people without obstructive sleep apnea, the rates in the general population, and the expectations according to chance. For each interval, we assessed the median apnea-hypopnea index and the relative risk of sudden death from cardiac causes. We similarly analyzed sudden death from cardiac causes during three time intervals that correlate with usual sleep-wake cycles. RESULTS: From midnight to 6 a.m., sudden death from cardiac causes occurred in 46 percent of people with obstructive sleep apnea, as compared with 21 percent of people without obstructive sleep apnea (P=0.01), 16 percent of the general population (P<0.001), and the 25 percent expected by chance (P<0.001). People with sudden death from cardiac causes from midnight to 6 a.m. had a significantly higher apnea-hypopnea index than those with sudden death from cardiac causes during other intervals, and the apnea-hypopnea index correlated directly with the relative risk of sudden death from cardiac causes from midnight to 6 a.m. For people with obstructive sleep apnea, the relative risk of sudden death from cardiac causes from midnight to 6 a.m. was 2.57 (95 percent confidence interval, 1.87 to 3.52). The analysis of usual sleep-wake cycles showed similar results. CONCLUSIONS: People with obstructive sleep apnea have a peak in sudden death from cardiac causes during the sleeping hours, which contrasts strikingly with the nadir of sudden death from cardiac causes during this period in people without obstructive sleep apnea and in the general population. PMID- 15788496 TI - ACVBP versus CHOP plus radiotherapy for localized aggressive lymphoma. AB - BACKGROUND: Chemoradiotherapy is standard treatment for localized aggressive lymphoma. To determine the optimal therapy for nonelderly persons with low-risk localized lymphoma, we conducted a randomized trial comparing chemoradiotherapy with chemotherapy alone. METHODS: Previously untreated patients less than 61 years old with localized stage I or II aggressive lymphoma and no adverse prognostic factors according to the International Prognostic Index were randomly assigned to three cycles of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) plus involved-field radiotherapy (329 patients) or chemotherapy alone with dose-intensified doxorubicin, cyclophosphamide, vindesine, bleomycin, and prednisone (ACVBP) plus sequential consolidation (318 patients). RESULTS: With a median follow-up of 7.7 years, event-free and overall survival rates were significantly higher in the group given chemotherapy alone than in the group given CHOP plus radiotherapy (P<0.001 and P=0.001, respectively). The five-year estimates of event-free survival were 82 percent (95 percent confidence interval, 78 to 87 percent) for patients receiving chemotherapy alone and 74 percent (95 percent confidence interval, 69 to 78 percent) for those receiving chemoradiotherapy. The respective five-year estimates of overall survival were 90 percent (95 percent confidence interval, 87 to 93 percent) and 81 percent (95 percent confidence interval, 77 to 86 percent). In a multivariate analysis, event free and overall survival rates were affected by treatment group, independently of tumor stage and the presence or absence of bulky disease. CONCLUSIONS: In patients under 61 years of age, chemotherapy with three cycles of ACVBP followed by sequential consolidation is superior to three cycles of CHOP plus radiotherapy for the treatment of low-risk localized lymphoma. PMID- 15788498 TI - Clinical practice. Pertussis--not just for kids. PMID- 15788499 TI - Polycystic ovary syndrome. PMID- 15788500 TI - Images in clinical medicine. Renal papillary necrosis unmasking sickle cell disease. PMID- 15788501 TI - Case records of the Massachusetts General Hospital. Case 9-2005. A 67-year-old man with acute respiratory failure. PMID- 15788502 TI - Defining the stages of aggressive non-Hodgkin's lymphoma--a work in progress. PMID- 15788503 TI - Financing health care--finding the money is hard and spending it well is even harder. PMID- 15788504 TI - Health care vouchers--a proposal for universal coverage. PMID- 15788505 TI - Do we really want broad access to health care? PMID- 15788506 TI - Palifermin and chemotherapy-induced oral mucositis. PMID- 15788507 TI - Care of the potential organ donor. PMID- 15788508 TI - Contagious acute gastrointestinal infections. PMID- 15788509 TI - Cancer of the ovary. PMID- 15788510 TI - Case 37-2004: postmenopausal bleeding and a cystic ovarian mass. PMID- 15788511 TI - Why "why" matters. PMID- 15788512 TI - Nosocomial transmission of cryptococcosis. PMID- 15788514 TI - Remapping the remembered target location for anti-saccades in human posterior parietal cortex. AB - We used functional magnetic resonance imaging (fMRI) to investigate the role of the human posterior parietal cortex (PPC) in anti-saccades. To do so, we exploited the laterality of a subregion of the PPC for remembered target location. Using an event-related design, we tracked fMRI signal changes in this region while subjects remembered the location of a flashed target, then were instructed to plan either an anti- or pro-saccade to that location, and finally were instructed to execute the movement. At first, the region responded preferentially to the memory of a target location presented in the contralateral visual field. However, when an anti-cue specified a saccadic response into the opposite visual field, we observed a dynamic shift in cortical activity from one hemisphere to the other. This shows that this region within the human posterior parietal cortex codes the target location for an upcoming saccade, rather than the location of the remembered visual stimulus in an anti-saccade task. PMID- 15788513 TI - Primate memory saccade amplitude after intervened motion depends on target distance. AB - To keep a stable internal representation of the visual world as our eyes, head, and body move around, humans and monkeys must continuously adjust neural maps of visual space using extraretinal sensory or motor cues. When such movements include translation, the amount of body displacement must be weighted differently in the updating of far versus near targets. Using a memory-saccade task, we have investigated whether nonhuman primates can benefit from this geometry when passively moved sideways. We report that monkeys made appropriate memory saccades, taking into account not only the amplitude and nature (rotation vs. translation) of the movement, but also the distance of the memorized target: i.e., the amplitude of memory saccades was larger for near versus far targets. The scaling by viewing distance, however, was less than geometrically required, such that memory saccades consistently undershot near targets. Such a less-than ideal scaling of memory saccades is reminiscent of the viewing distance-dependent properties of the vestibuloocular reflex. We propose that a similar viewing distance-dependent vestibular signal is used as an extraretinal compensation for the visuomotor consequences of the geometry of motion parallax by scaling both memory saccades and reflexive eye movements during motion through space. PMID- 15788515 TI - Electrical coupling in sustentacular cells of the mouse olfactory epithelium. AB - Sustentacular cells (SCs) line the apical surface of the olfactory epithelium (OE) and provide trophic, metabolic, and mechanical support for olfactory receptor neurons. Morphological studies have suggested that SCs possess gap junctions, although physiological evidence for gap junctional communication in mammalian SCs is lacking. In the present study we investigated whether coupling exists between SCs situated in tissue slices of OE from neonatal (P0-P4) mice. Using whole cell and cell-attached patch recordings from SCs, we demonstrate that SCs are electrically coupled by junctional resistances on the order of 300 M(omega). Under whole cell recording conditions, Alexa 488 added to the pipette solution failed to reveal dye coupling between SCs. Electrical coupling was deduced from the biexponential decay of capacitive currents recorded from SCs and from the bell-shaped voltage dependency of a P2Y-receptor-activated current, both of which were abolished by 18beta-glycyrrhetinic acid (20-50 microM), a blocker of gap junctions. These data provide strong evidence for functional coupling between SCs, the physiological importance of which is discussed. PMID- 15788516 TI - Analysis of 5-HT-induced short-term facilitation at Aplysia sensorimotor synapse during bursts: increased synaptic gain that does not require ERK activation. AB - The 5-HT-induced synaptic plasticity of Aplysia sensorimotor synapses has typically been probed by firing a single presynaptic spike. In this study, 5-HT induced synaptic plasticity was probed with brief bursts of spikes (10 Hz, 1 s), which are more behaviorally relevant stimuli. Because such bursts provide a greater challenge to the release machinery than single spikes, their use may reveal additional aspects of synaptic modulation, and, in particular, the role of extracellular signal-regulated protein kinase (ERK), which has recently been implicated in several examples of short- and long-term synaptic plasticity. Excitatory postsynaptic currents (EPSCs) were characterized by their amplitudes. In addition, two kinetic measurements, time to peak and decay time constant, were determined for the initial and last EPSCs of each burst. Application of 5-HT produced a uniform increase in gain by facilitating each EPSC elicited during a burst of spikes without affecting the kinetics of the initial or last EPSC. These data suggest that short-term facilitation during a burst is mediated largely by processes such as those that affect the size of the releasable pool or rate of vesicle mobilization rather than by an increase in the duration of the presynaptic action potential. An ERK cascade inhibitor (U0126) had no effect on the 5-HT-mediated facilitation of either the initial EPSC or EPSCs elicited late in the burst. PMID- 15788517 TI - Motor cortical modulation of cutaneous reflex responses in the hindlimb of the intact cat. AB - We have used the technique of spatial facilitation to examine the interactions between the signals conveyed by the corticospinal tract and those of cutaneous afferents in the hindlimb of the intact, walking cat. Microstimulation was applied to 20 cortical sites in the hindlimb representation of the motor cortex and to three different cutaneous nerves innervating the hindpaw in four cats. Conditioning stimuli to the motor cortex induced both facilitation and depression of cutaneous reflexes evoked by stimulation of nerves in the hindlimb contralateral to the cortical stimulation site. Facilitation was most frequently evoked by conditioning stimuli in the range of 10-30 ms before the cutaneous stimulation; depression was normally evoked by shorter and longer conditioning delays. Similar changes were observed after conditioning stimuli to the pyramidal tract, suggesting that the changes were independent of any changes in cortical excitability. Modulation of reflex activity varied according to the muscle under study, the cutaneous nerve used to evoke the reflex and the cortical site used to condition the reflex. Together, these results suggest that there is spatial convergence of corticospinal and cutaneous afferent activity and that this convergence is mediated by distinct subpopulations of spinal interneurons. PMID- 15788518 TI - Contribution of the motor cortex to the structure and the timing of hindlimb locomotion in the cat: a microstimulation study. AB - We used microstimulation to examine the contribution of the motor cortex to the structure and timing of the hindlimb step cycle during locomotion in the intact cat. Stimulation was applied to the hindlimb representation of the motor cortex in 34 sites in three cats using either standard glass-insulated microelectrodes (16 sites in 1 cat) or chronically implanted microwire electrodes (18 sites in 2 cats). Stimulation at just suprathreshold intensities with the cat at rest produced multi-joint movements at a majority of sites (21/34, 62%) but evoked responses restricted to a single joint, normally the ankle, at the other 13/34 (38%) sites. Stimulation during locomotion generally evoked larger responses than the same stimulation at rest and frequently activated additional muscles. Stimulation at all 34 sites evoked phase-dependent responses in which stimulation in swing produced transient increases in activity in flexor muscles while stimulation during stance produced transient decreases in activity in extensors. Stimulation with long (200 ms) trains of stimuli in swing produced an increased level of activity and duration of flexor muscles without producing changes in cycle duration. In contrast, stimulation during stance decreased the duration of the extensor muscle activity and initiated a new and premature period of swing, resetting the step cycle. Stimulation of the pyramidal tract in two of these three cats as well as in two additional ones produced similar effects. The results show that the motor cortex is capable of influencing hindlimb activity during locomotion in a similar manner to that seen for the forelimb. PMID- 15788519 TI - Modeling the effects of transcranial magnetic stimulation on cortical circuits. AB - Transcranial magnetic stimulation (TMS) is commonly used to activate or inactivate specific cortical areas in a noninvasive manner. Because of technical constraints, the precise effects of TMS on cortical circuits are difficult to assess experimentally. Here, this issue is investigated by constructing a detailed model of a portion of the thalamocortical system and examining the effects of the simulated delivery of a TMS pulse. The model, which incorporates a large number of physiological and anatomical constraints, includes 33,000 spiking neurons arranged in a 3-layered motor cortex and over 5 million intra- and interlayer synaptic connections. The model was validated by reproducing several results from the experimental literature. These include the frequency, timing, dose response, and pharmacological modulation of epidurally recorded responses to TMS (the so-called I-waves), as well as paired-pulse response curves consistent with data from several experimental studies. The modeled responses to simulated TMS pulses in different experimental paradigms provide a detailed, self consistent account of the neural and synaptic activities evoked by TMS within prototypical cortical circuits. PMID- 15788520 TI - Mg2+-malate co-transport, a mechanism for Na+-independent Mg2+ transport in neurons of the leech Hirudo medicinalis. AB - Mg2+-extrusion from Mg2+-loaded neurons of the leech, Hirudo medicinalis, is mediated mainly by Na+/Mg2+ antiport. However, in a number of leech neurons, Mg2+ is extruded in the nominal absence of extracellular Na+, indicating the existence of an additional, Na+-independent Mg2+ transport mechanism. This mechanism was investigated using electrophysiological and microfluorimetrical techniques. The rate of Na+-independent Mg2+ extrusion from Mg2+-loaded leech neurons was found to be independent of extracellular Ca2+, K+, NO3-, HCO3-, SO4(2-), HPO4(2-), and of intra- and extracellular pH. Na+-independent Mg2+ extrusion was not inhibited by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), furosemide, ouabain, vanadate, iodoacetate, 4-amino-hippurate, or alpha-cyano-4 hydroxycinnamate and was not influenced by changes in the membrane potential in voltage-clamp experiments. Na+-independent Mg2+ extrusion was, however, inhibited by the application of 2 mM probenecid, a blocker of organic anion transporters, suggesting that Mg2+ might be co-transported with organic anions. Extracellularly, of all organic anions tested (malate, citrate, lactate, alpha ketoglutarate, and 4-amino-hippurate) only high, but physiological, concentrations of malate (30 mM) had a significant inhibitory effect on Na+ independent Mg2+ extrusion. Intracellularly, iontophoretically injected malate, citrate, or fura-2, but not Cl-, alpha-ketoglutarate, glutamate, succinate, or urate, were stimulating Na+-independent Mg2+ extrusion from those neurons that initially did not extrude Mg2+ in Na+-free solutions. Our data indicate that Mg2+ is co-transported with organic anions, preferably with malate, the predominant extracellular anion in the leech. The proposed model implies that, under experimental conditions, malate drives Mg2+ extrusion, whereas under physiological conditions, malate is actively taken up, driven by Mg2+, so that malate can be metabolized. PMID- 15788521 TI - Effect of interocular delay on disparity-selective v1 neurons: relationship to stereoacuity and the pulfrich effect. AB - The temporal properties of disparity-sensitive neurons place important temporal constraints on stereo matching. We examined these constraints by measuring the responses of disparity-selective neurons in striate cortex of awake behaving monkeys to random-dot stereograms that contained interocular delays. Disparity selectivity was gradually abolished by increasing interocular delay (when the delay exceeds the integration time, the inputs from the 2 eyes become uncorrelated). The amplitude of the disparity-selective response was a Gaussian function of interocular delay, with a mean of 16 ms (+/-5 ms, SD). Psychophysical measures of stereoacuity, in both monkey and human observers, showed a closely similar dependency on time, suggesting that temporal integration in V1 neurons is what determines psychophysical matching constraints over time. There was a slight but consistent asymmetry in the neuronal responses, as if the optimum stimulus is one in which the right stimulus leads by about 4 ms. Because all recordings were made in the left hemisphere, this probably reflects nasotemporal differences in conduction times; psychophysical data are compatible with this interpretation. In only a few neurons (5/72), interocular delay caused a change in the preferred disparity. Such tilted disparity/delay profiles have been invoked previously to explain depth perception in the stroboscopic version of the Pulfrich effect (and other variants). However, the great majority of the neurons did not show tilted disparity/delay profiles. This suggests that either the activity of these neurons is ignored when viewing Pulfrich stimuli, or that current theories relating neuronal properties to perception in the Pulfrich effect need to be reevaluated. PMID- 15788522 TI - Pitch of complex tones: rate-place and interspike interval representations in the auditory nerve. AB - Harmonic complex tones elicit a pitch sensation at their fundamental frequency (F0), even when their spectrum contains no energy at F0, a phenomenon known as "pitch of the missing fundamental." The strength of this pitch percept depends upon the degree to which individual harmonics are spaced sufficiently apart to be "resolved" by the mechanical frequency analysis in the cochlea. We investigated the resolvability of harmonics of missing-fundamental complex tones in the auditory nerve (AN) of anesthetized cats at low and moderate stimulus levels and compared the effectiveness of two representations of pitch over a much wider range of F0s (110-3,520 Hz) than in previous studies. We found that individual harmonics are increasingly well resolved in rate responses of AN fibers as the characteristic frequency (CF) increases. We obtained rate-based estimates of pitch dependent upon harmonic resolvability by matching harmonic templates to profiles of average discharge rate against CF. These estimates were most accurate for F0s above 400-500 Hz, where harmonics were sufficiently resolved. We also derived pitch estimates from all-order interspike-interval distributions, pooled over our entire sample of fibers. Such interval-based pitch estimates, which are dependent on phase-locking to the harmonics, were accurate for F0s below 1,300 Hz, consistent with the upper limit of the pitch of the missing fundamental in humans. The two pitch representations are complementary with respect to the F0 range over which they are effective; however, neither is entirely satisfactory in accounting for human psychophysical data. PMID- 15788523 TI - Object, space, and object-space representations in the primate hippocampus. AB - A fundamental question about the function of the primate including human hippocampus is whether object as well as allocentric spatial information is represented. Recordings were made from single hippocampal formation neurons while macaques performed an object-place memory task that required the monkeys to learn associations between objects and where they were shown in a room. Some neurons (10%) responded differently to different objects independently of location; other neurons (13%) responded to the spatial view independently of which object was present at the location; and some neurons (12%) responded to a combination of a particular object and the place where it was shown in the room. These results show that there are separate as well as combined representations of objects and their locations in space in the primate hippocampus. This is a property required in an episodic memory system, for which associations between objects and the places where they are seen are prototypical. The results thus provide an important advance by showing that a requirement for a human episodic memory system, separate and combined neuronal representations of objects and where they are seen "out there" in the environment, is present in the primate hippocampus. PMID- 15788524 TI - Fully tuneable stochastic resonance in cutaneous receptors. AB - Stochastic resonance describes a phenomenon whereby the addition of "noise" to the input of a nonlinear system can improve sensitivity. "Fully tuneable stochastic resonance" is a particular form of the phenomenon that requires the matching of two time scales: one being that of the subthreshold periodic stimulus of the system and the other being the noise-induced response of the system. First proposed in 1981, stochastic resonance has been reported in a wide range of biological systems; however, conclusive experimental evidence for fully tuneable stochastic resonance in biological systems is limited. Evidence of fully tuneable stochastic resonance in the response of slowly adapting type I mechanoreceptors in the toad is presented. The results are extended to include the first evidence supporting fully tuneable stochastic resonance in psychophysical experiments, namely tactile detection thresholds, indicating that the human CNS is capable of accessing the improved information available via fully tuneable stochastic resonance. PMID- 15788525 TI - Building capacity for health promotion--a case study from China. AB - During the period 1997-2000 a technical assistance project to build capacity for community-based health promotion was implemented in seven cities and one province in China. The technical assistance project formed part of a much larger World Bank supported program to improve disease prevention capabilities in China, commonly known as Health VII. The technical assistance project was funded by the Australian Agency for International Development. It was designed to develop capacity within the Ministry of Health (MOH) and the cities and province in the management of community-based health promotion projects, as well as supporting institutional development and public health policy reform. There are some relatively unique features of this technical assistance which helped shape its implementation and impact. It sought to provide the Chinese MOH and the cities and province with an introduction to comprehensive health promotion strategies, in contrast to the more limited information, education and communication strategies. The project was provided on a continuing basis over 3 years through a single institution, rather than as a series of ad hoc consultancies by individuals. Teaching and learning processes were developmental, leading progressively to a greater degree of local Chinese input and management to ensure sustainability and maintenance of technical support for the project. Based on this experience, this paper presents a model for capacity building projects of this type. It describes the education, training and planning activities that were the key inputs to the project, as well as the limited available evidence on the impact of the project. It describes how the project evolved over time to meet the changing needs of the participants, specifically how the content of the project shifted from a risk-factor orientation to a settings-based focus, and the delivery of the project moved from an expert-led approach to a more participatory, problem based learning approach. In terms of impact, marked differences before and after the implementation of the training activities were identified in key areas for reform, in addition to the self reported positive change in knowledge, and a high level of participant satisfaction. Key lessons are summarized. Technical assistance projects of this kind benefit from continuity and a high level of coordination, the provision of culturally and linguistically appropriate teaching, and a clear understanding of the need to match workforce development with organizational/institutional development. PMID- 15788526 TI - Understanding health literacy: an expanded model. AB - A long and yet unfinished history of investigating how individual capabilities and social processes explain or predict health indicates that poor education, low literacy, poor health and early death are strongly linked around the world. However, the complexity of those relationships is not fully understood. In this article, we propose an expanded model of health literacy characterized by four domains: fundamental literacy (reading, writing, speaking and numeracy), science literacy, civic literacy and cultural literacy. To explore the utility of this model, we examine selected pieces of the public discourse about terrorism and bioterrorism that dominated the mass media during the anthrax threat in the United States during 2001. We conclude that this model of health literacy is useful to analyze health communication, to aid in constructing more understandable and appropriate health communication, and ultimately can lead to the development of a new measure to assess health literacy skills in individuals. PMID- 15788527 TI - Evaluating health-promoting schools in Hong Kong: development of a framework. AB - Health-promoting schools (HPS)/healthy schools have existed internationally for about 15 years. Yet there are few comprehensive evaluation frameworks available which enable the outcomes of HPS initiatives to be assessed. This paper identifies an evaluation framework developed in Hong Kong. The framework uses a range of approaches to explore what schools actually do in their health promotion and health education initiatives. The framework, which is based on the WHO (Western Pacific Regional Office) Guidelines for HPS, is described in detail. The appropriate instruments for data collection are described and their origins identified. The evaluation plan and protocol, which underpinned the very comprehensive evaluation in Hong Kong, are explained. Finally, a case is argued for evaluation of HPS to be more in line with the educational dynamics of schools and the research literature on effective schooling, rather than focusing primarily on health-related measures. PMID- 15788528 TI - Improving physical fitness and emotional well-being in adolescents of low socioeconomic status in Chile: results of a school-based controlled trial. AB - Regular physical activity is associated with a reduced risk of all-cause mortality, and mortality due to cardiovascular disease and cancer. Among adolescents, physical activity is associated with benefits in the prevention and control of emotional distress, and improvement of self-esteem. Countries in transitional epidemiological scenarios, such as Chile, need to develop effective strategies to improve physical activity as a way to face the epidemic of chronic diseases. The objective of this study was to evaluate the effects of a school based physical activity program on physical fitness and mental health status of adolescents living in a low socioeconomic status area in Santiago, Chile. A quasi experimental design was used to evaluate the effects of the program over one academic year. The study included 198 students aged 15 years old. Two ninth grade classes were randomly selected as the intervention group, with two classes of the same grade as controls. A social planning approach was used to develop the intervention. The program was designed and implemented based on student preferences, teachers' expertise and local resources. Changes in physiological and mental health status were assessed. After the intervention, maximum oxygen capacity achieved a significant increase of 8.5% in the intervention versus 1.8% in the control group (p < 0.0001). Speed and jump performance scores improved significantly more in the intervention versus the control group (p > 0.01). Anxiety score decreased 13.7% in the intervention group versus 2.8% in the control group (p < 0.01), and self-esteem score increased 2.3% in the intervention group and decreased 0.1% in the control group after the end of the program (p < 0.0001). No significant change was observed in the depressive score. Student participation and compliance with the program was > 80%. To conclude, a school-based program to improve physical activity in adolescents of low socioeconomic status, obtained a high level of participation and achieved significant benefits in terms of physical fitness and mental health status. PMID- 15788529 TI - The complete genomes and proteomes of 27 Staphylococcus aureus bacteriophages. AB - Bacteriophages are the most abundant life forms in the biosphere. They play important roles in bacterial ecology, evolution, adaptation to new environments, and pathogenesis of human bacterial infections. Here, we report the complete genomic sequences, and predicted proteins of 27 bacteriophages of the Gram positive bacterium Staphylococcus aureus. Comparative nucleotide and protein sequence analysis indicates that these phages are a remarkable source of untapped genetic diversity, encoding 2,170 predicted protein-encoding ORFs, of which 1,402 cannot be annotated for structure or function, and 522 are proteins with no similarity to other phage or bacterial sequences. Based on their genome size, organization of their gene map and comparative nucleotide and protein sequence analysis, the S. aureus phages can be organized into three groups. Comparison of their gene maps reveals extensive genome mosaicism, hinting to a large reservoir of unidentified S. aureus phage genes. Among the phages in the largest size class (178-214 kbp) that we characterized is phage Twort, the first discovered bacteriophage (responsible for the Twort-D'Herelle effect). These phage genomes offer an exciting opportunity to discern molecular mechanisms of phage evolution and diversity. PMID- 15788530 TI - Contingent gene regulatory networks and B cell fate specification. AB - The B cell developmental pathway represents a leading system for the analysis of regulatory circuits that orchestrate cell fate specification and commitment. Considerable progress has been achieved within the past decade in the identification and genetic analysis of various regulatory components. These components include the transcription factors PU.1, Ikaros, Bcl11a, E2A, EBF, and Pax-5, as well as the cytokine receptors Flk2 and IL-7R. Experimental evidence of connectivity among the regulatory components is used to assemble sequentially acting and contingent gene regulatory networks. Transient signaling inputs, self sustaining positive feedback loops, and cross-antagonism among alternate cell fate determinants are key features of the proposed networks that instruct the development of B lymphocyte precursors from hematopoietic stem cells. PMID- 15788531 TI - Logic functions of the genomic cis-regulatory code. AB - cis-Regulatory modules that control developmental gene expression process the regulatory inputs provided by the transcription factors for which they contain specific target sites. A prominent class of cis-regulatory processing functions can be modeled as logic operations. Many of these are combinatorial because they are mediated by multiple sites, although others are unitary. In this work, we illustrate the repertoire of cis-regulatory logic operations, as an approach toward a functional interpretation of the genomic regulatory code. PMID- 15788532 TI - Dynamic microcompartmentation in synthetic cells. AB - An experimental model for cytoplasmic organization is presented. We demonstrate dynamic control over protein distribution within synthetic cells comprising a lipid bilayer membrane surrounding an aqueous polymer solution. This polymer solution generally exists as two immiscible aqueous phases. Protein partitioning between these phases leads to microcompartmentation, or heterogeneous protein distribution within the "cell" interior. This model cytoplasm can be reversibly converted to a single phase by slight changes in temperature or osmolarity, such that local protein concentrations can be manipulated within the vesicle interior. PMID- 15788533 TI - Structural origins of efficient proton abstraction from carbon by a catalytic antibody. AB - Antibody 34E4 catalyzes the conversion of benzisoxazoles to salicylonitriles with high rates and multiple turnovers. The crystal structure of its complex with the benzimidazolium hapten at 2.5-angstroms resolution shows that a combination of hydrogen bonding, pi stacking, and van der Waals interactions is exploited to position both the base, Glu(H50), and the substrate for efficient proton transfer. Suboptimal placement of the catalytic carboxylate, as observed in the 2.8-angstroms structure of the Glu(H50)Asp variant, results in substantially reduced catalytic efficiency. In addition to imposing high positional order on the transition state, the antibody pocket provides a highly structured microenvironment for the reaction in which the carboxylate base is activated through partial desolvation, and the highly polarizable transition state is stabilized by dispersion interactions with the aromatic residue Trp(L91) and solvation of the leaving group oxygen by external water. The enzyme-like efficiency of general base catalysis in this system directly reflects the original hapten design, in which a charged guanidinium moiety was strategically used to elicit an accurately positioned functional group in an appropriate reaction environment and suggests that even larger catalytic effects may be achievable by extending this approach to the induction of acid-base pairs capable of bifunctional catalysis. PMID- 15788534 TI - Leptin increase in multiple sclerosis associates with reduced number of CD4(+)CD25+ regulatory T cells. AB - We analyzed the serum and cerebrospinal fluid (CSF) leptin secretion and the interaction between serum leptin and CD4(+)CD25+ regulatory T cells (T(Regs)) in naive-to-therapy relapsing-remitting multiple sclerosis (RRMS) patients. Leptin production was significantly increased in both serum and CSF of RRMS patients and correlated with IFN-gamma secretion in the CSF. T cell lines against human myelin basic protein (hMBP) produced immunoreactive leptin and up-regulated the expression of the leptin receptor (ObR) after activation with hMBP. Treatment with either anti-leptin or anti-leptin-receptor neutralizing antibodies inhibited in vitro proliferation in response to hMBP. Interestingly, in the RRMS patients, an inverse correlation between serum leptin and percentage of circulating T(Regs) was also observed. To better analyze the finding, we enumerated T(Regs) in leptin deficient (ob/ob) and leptin-receptor-deficient (db/db) mice and observed the significant increase in T(Regs). Moreover, treatment of WT mice with soluble ObR fusion protein (ObR:Fc) increased the percentage of T(Regs) and ameliorated the clinical course and progression of disease in proteolipid protein peptide (PLP(139-151))-induced relapsing-experimental autoimmune encephalomyelitis (R EAE), an animal model of RRMS. These findings show an inverse relationship between leptin secretion and the frequency of T(Regs) in RRMS and may have implications for the pathogenesis of and therapy for multiple sclerosis. PMID- 15788535 TI - Regulation of hematopoietic stem cell aging in vivo by a distinct genetic element. AB - Until recently, stem cells were thought to be endowed with unlimited self-renewal capacity and, thus, assumed exempt from aging. But accumulating evidence over the past decade compellingly argues that a measurable and progressive replicative impairment in the hematopoietic, intestinal, and muscle stem cell activity exists from adulthood to old age, resulting in a decline in stem cell function and rendering stem cell aging as the possible link between cellular aging and organismal aging. By using a previously uncharacterized congenic animal model to study genetic regulation of hematopoietic stem cell aging, we have demonstrated definitively that a locus on murine chromosome 2 regulates hematopoietic stem cell aging. In addition to demonstrating that hematopoietic stem cell aging is regulated by a distinct genetic element, experimental evidence links the response of hematopoietic stem cells to DNA double-strand breaks to cellular aging, suggesting DNA integrity influences stem cell aging. PMID- 15788536 TI - Phosphorylation of Hdmx mediates its Hdm2- and ATM-dependent degradation in response to DNA damage. AB - Maintenance of genomic stability depends on the DNA damage response, an extensive signaling network that is activated by DNA lesions such as double-strand breaks (DSBs). The primary activator of the mammalian DSB response is the nuclear protein kinase ataxia-telangiectasia, mutated (ATM), which phosphorylates key players in various arms of this network. The activation and stabilization of the p53 protein play a major role in the DNA damage response and are mediated by ATM dependent posttranslational modifications of p53 and Mdm2, a ubiquitin ligase of p53. p53's response to DNA damage also depends on Mdm2-dependent proteolysis of Mdmx, a homologue of Mdm2 that represses p53's transactivation function. Here we show that efficient damage-induced degradation of human Hdmx depends on functional ATM and at least three sites on the Hdmx that are phosphorylated in response to DSBs. One of these sites, S403, is a direct ATM target. Accordingly, each of these sites is important for Hdm2-mediated ubiquitination of Hdmx after DSB induction. These results demonstrate a sophisticated mechanism whereby ATM fine-tunes the optimal activation of p53 by simultaneously modifying each player in the process. PMID- 15788537 TI - Gene regulatory networks for development. AB - The genomic program for development operates primarily by the regulated expression of genes encoding transcription factors and components of cell signaling pathways. This program is executed by cis-regulatory DNAs (e.g., enhancers and silencers) that control gene expression. The regulatory inputs and functional outputs of developmental control genes constitute network-like architectures. In this PNAS Special Feature are assembled papers on developmental gene regulatory networks governing the formation of various tissues and organs in nematodes, flies, sea urchins, frogs, and mammals. Here, we survey salient points of these networks, by using as reference those governing specification of the endomesoderm in sea urchin embryos and dorsal-ventral patterning in the Drosophila embryo. PMID- 15788538 TI - Global energy gradients and size in colonial organisms: worker mass and worker number in ant colonies. AB - Body mass shapes processes from cell metabolism to community dynamics. Little is known, however, about how the average body mass of individuals varies among ecological communities. Ants alter colony mass by independently changing worker mass and/or worker number. In a survey of 49 ecosystems from tundra to tropical rainforest, average worker mass and worker number were uncorrelated (r(s) = 0.2, P > 0.14) and varied 100-fold. Data supported the hypothesis that higher mean monthly temperatures, T, reduce worker mass by increasing metabolic costs during worker development. In contrast, worker number was unimodal over a 1,000-fold gradient of net primary productivity (NPP, g of carbon per m2 per yr), a measure of organic carbon available to consumers. At the lowest levels of NPP colonies appeared to be carbon-limited; above 60 g of carbon per m2 per yr average worker number decreased to a global low. This decline in worker number with increasing NPP supports the hypothesis that abundant carbon ameliorates the Achilles heel of small taxa in competition with large taxa: their relatively high metabolic demands. Higher predation rates in resource-rich environments may also play a role in limiting worker number. In all, about half the global variation in worker mass and number was accounted for by gradients of NPP and T. Changes in global temperature and rainfall may thus mold gradients of ectotherm size, with consequences for the structure and function of the ecosystems. PMID- 15788539 TI - Determination of drug glucuronidation and UDP-glucuronosyltransferase selectivity using a 96-well radiometric assay. AB - A rapid and sensitive radiometric assay for UDP-glucuronosyltransferase (UGT) is described. UGT substrates are incubated in 96-well plates with microsomes in the presence of [14C]UDP-glucuronic acid, and 14C-labeled glucuronidation products are separated from the unreacted nucleotide sugar by solid-phase extraction using 96-well extraction plates. The assay was validated with 15 structurally diverse UGT substrates containing acidic, phenolic, and hydroxyl reacting groups. Glucuronidation velocities for these compounds were determined using human, rat, and dog liver microsomes, and reaction kinetics were studied with 1-naphthol and 4-methylumbelliferone. Results obtained with the new assay confirmed the previously reported rank order of glucuronidation velocity of several typical UGT substrates and the finding that the glucuronidation of most of these compounds is significantly faster in dog than in human liver microsomes. UGT specificity of five compounds was determined using recombinant human UGTs. The major UGT isoforms identified were UGT1A6, UGT1A7, and UGT1A9 for 4-methylumbelliferone; UGT1A6 and UGT1A8 for 1-naphthol; UGT2B7 for naloxone; UGT1A3 and UGT2B7 for ketoprofen; and UGT1A4 for trifluoperazine. Identical results were obtained with a conventional high-performance liquid chromatography method coupled to mass spectrometric detection. The new assay should prove valuable for rapidly benchmarking recombinant UGTs and microsomal preparations from different species and tissues, identifying high-turnover compounds during drug discovery, and reaction phenotyping studies. PMID- 15788540 TI - In vitro-in vivo extrapolation of CYP2D6 inactivation by paroxetine: prediction of nonstationary pharmacokinetics and drug interaction magnitude. AB - Attempts at predicting drug-drug interactions perpetrated by paroxetine from in vitro data have utilized reversible enzyme inhibition models and have been unsuccessful to date, grossly underpredicting interaction magnitude. Recent data have provided evidence for mechanism-based inactivation of CYP2D6 by paroxetine. We have predicted the pharmacokinetic consequences of CYP2D6 inactivation by paroxetine from in vitro inactivation kinetics (kinact 0.17 min(-1), unbound KI 0.315 microM), in vivo inhibitor concentrations, and an estimated CYP2D6 degradation half-life of 51 h, using a mathematical model of mechanism-based inhibition. The model-predicted accumulation ratio of paroxetine was 5 times that expected from single-dose kinetics and in excellent agreement with the observed 5 to 6-fold greater accumulation. Magnitudes of interactions produced by paroxetine (20-30 mg/day) with desipramine, risperidone, perphenazine, atomoxetine, (S)-metoprolol, and (R)-metoprolol were predicted, considering the contribution of CYP2D6 to their oral clearance. Predicted fold-increases in victim drug AUC were 5-, 6-, 5-, 6-, 4-, and 6-fold, respectively, and are in reasonable agreement with observed values of 5-, 6-, >7-, 7-, 5-, and 8-fold, respectively. Failure to consider microsomal binding in vitro adversely affected predictive accuracy. Simulation of the sensitivities of these predictions to model inputs suggests a 2-fold underprediction of interaction magnitude when a CYP2D6 degradation half-life of 14 h (reported for rat CYP3A) is used. In summary, the scaling model for mechanism-based inactivation successfully predicted the pharmacokinetic consequences of CYP2D6 inactivation by paroxetine from in vitro data. PMID- 15788541 TI - Change in challenging times: a plan for extending and improving health coverage. AB - Some speculate that Americans are neither politically capable of nor morally committed to solving the health system problems. We disagree. We propose a plan that insures all and improves the value and cost-effectiveness of health care by knitting together employer-sponsored insurance and Medicaid; promoting prevention, research, and information technology; and financing its investments through a dedicated value-added tax. By prioritizing practicality, fairness, and responsibility, the plan aims to avoid ideological battles and prevent fear of major change. By emphasizing the moral imperative for change, especially relative to other options on the policy agenda, it aims to create momentum for expanding and improving health coverage for all. PMID- 15788543 TI - Cognitive and social impairments in patients with superficial siderosis. AB - Superficial siderosis of the CNS is a rare condition, caused by deposition of haemosiderin in the superficial layers of the CNS due to repeated chronic subarachnoid or intraventricular haemorrhage. Typically, the hindbrain structures, especially the cerebellum, are most affected. There is a surprising lack of studies investigating in detail the behavioural functioning of patients with such a condition. In this study, we document for the first time the cognitive, social and emotional processing of six patients with a confirmed clinical diagnosis of superficial siderosis. They were aged between 40 and 62 years, with a mean age of 50.2 years; four were male. We administered a comprehensive battery of general cognitive ability and social cognitive tasks. A review of MRI was also undertaken. The findings indicate selective cognitive impairments affecting speech production, visual recall memory and executive functions. In addition, a selective pattern of social dysfunction, affecting the ability to represent other people's mental states, was found. These behavioural dysfunctions are reported in the context of MRI-documented lesions maximally involving the cerebellum, in particular the superior vermis, as well as the medial and inferior frontal cortex. These results suggest that superficial siderosis is associated with a distinct pattern of cognitive and social impairments. They are consistent with the recently proposed role of the cerebellum as a modulator of cognitive, social and emotional functions. PMID- 15788542 TI - Characteristics of two distinct clinical phenotypes in pathologically proven progressive supranuclear palsy: Richardson's syndrome and PSP-parkinsonism. AB - The clinical diagnosis of progressive supranuclear palsy (PSP) relies on the identification of characteristic signs and symptoms. A proportion of pathologically diagnosed cases do not develop these classic features, prove difficult to diagnose during life and are considered as atypical PSP. The aim of this study was to examine the apparent clinical dichotomy between typical and atypical PSP, and to compare the biochemical and genetic characteristics of these groups. In 103 consecutive cases of pathologically confirmed PSP, we have identified two clinical phenotypes by factor analysis which we have named Richardson's syndrome (RS) and PSP-parkinsonism (PSP-P). Cases of RS syndrome made up 54% of all cases, and were characterized by the early onset of postural instability and falls, supranuclear vertical gaze palsy and cognitive dysfunction. A second group of 33 (32%) were characterized by asymmetric onset, tremor, a moderate initial therapeutic response to levodopa and were frequently confused with Parkinson's disease (PSP-P). Fourteen cases (14%) could not be separated according to these criteria. In RS, two-thirds of cases were men, whereas the sex distribution in PSP-P was even. Disease duration in RS was significantly shorter (5.9 versus 9.1 years, P < 0.001) and age at death earlier (72.1 versus 75.5 years, P = 0.01) than in PSP-P. The isoform composition of insoluble tangle-tau isolated from the basal pons also differed significantly. In RS, the mean four-repeat:three-repeat tau ratio was 2.84 and in PSP-P it was 1.63 (P < 0.003). The effect of the H1,H1 PSP susceptibility genotype appeared stronger in RS than in PSP-P (odds ratio 13.2 versus 4.5). The difference in genotype frequencies between the clinical subgroups was not significant. There were no differences in apolipoprotein E genotypes. The classic clinical description of PSP, which includes supranuclear gaze palsy, early falls and dementia, does not adequately describe one-third of cases in this series of pathologically confirmed cases. We propose that PSP-P represents a second discrete clinical phenotype that needs to be clinically distinguished from classical PSP (RS). The different tau isoform deposition in the basal pons suggests that this may ultimately prove to be a discrete nosological entity. PMID- 15788544 TI - The role of the striatum in rule application: the model of Huntington's disease at early stage. AB - The role of the basal ganglia, and more specifically of the striatum, in language is still debated. Recent studies have proposed that linguistic abilities involve two distinct types of processes: the retrieving of stored information, implicating temporal lobe areas, and the application of combinatorial rules, implicating fronto-striatal circuits. Studies of patients with focal lesions and neurodegenerative diseases have suggested a role for the striatum in morphological rule application, but functional imaging studies found that the left caudate was involved in syntactic processing and not morphological processing. In the present study, we tested the view that the basal ganglia are involved in rule application and not in lexical retrieving in a model of striatal dysfunction, namely Huntington's disease at early stages. We assessed the rule lexicon dichotomy in the linguistic domain with morphology (conjugation of non verbs and verbs) and syntax (sentence comprehension) and in a non-linguistic domain with arithmetic operations (subtraction and multiplication). Thirty Huntington's disease patients (15 at stage I and 15 at stage II) and 20 controls matched for their age and cultural level were included in this study. Huntington's disease patients were also assessed using the Unified Huntington's Disease Rating Scale (UHDRS) and MRI. We found that early Huntington's disease patients were impaired in rule application in the linguistic and non-linguistic domains (morphology, syntax and subtraction), whereas they were broadly spared with lexical processing. The pattern of performance was similar in patients at stage I and stage II, except that stage II patients were more impaired in all tasks assessing rules and had in addition a very slight impairment in the lexical condition of conjugation. Finally, syntactic rule abilities correlated with all markers of the disease evolution including bicaudate ratio and performance in executive function, whereas there was no correlation with arithmetic and morphological abilities. Together, this suggests that the striatum is involved in rule processing more than in lexical processing and that it extends to linguistic and non-linguistic domains. These results are discussed in terms of domain specific versus domain-general processes of rule application. PMID- 15788545 TI - Occipital epilepsy: lateral versus mesial. AB - This study compares ictal semiology, neurological examination and scalp EEG between lateral and mesial occipital epilepsy to assess the contribution non invasive data make in determining the epileptogenic region within an occipital lobe. We assessed seizure origin in 41 occipital patients as lateral (11 patients), mesial (20) and both surfaces (10) as indicated by subdurally recorded seizures (nine), a lesion whose removal reduced seizure quantity by > or =90% (six), or who met both criteria (26). No aspect of semiology distinguished lateral from mesially originating occipital seizures. A pre-operative visual field deficit appeared in eight (42%) out of 19 testable patients with mesial originating seizures, three (30%) out of 10 patients with both surfaces epileptogenic, but none of the 10 testable patients whose seizures arose only from the lateral surface (P = 0.0373, lateral versus mesial and both surfaces). Although occipital seizures appeared on the majority of the first five scalp EEG recordings in four (36%) out of 11 patients with laterally originating occipital seizures compared with none of 20 patients in whom seizures originated mesially (P = 0.0105), no other scalp EEG feature distinguished seizures from these surfaces. We conclude that subdural electroencephalography is likely to be necessary to delineate the epileptogenic region within an occipital lobe. Nonetheless, focally originating scalp-recorded seizures accurately lateralized the epileptogenic zone in 20 (49%) of our 41 patients compared with only one (2%) which originated contralaterally (P = 0.0001). This relationship held when considering only the first five scalp EEGs: the seizures of 10 patients (24%) appeared ipsilaterally and none contralaterally (P = 0.001). Moreover, interictal occipital (01,2) and posterior temporal (T5, T6) spikes appeared consistently and significantly (P < 0.001) more commonly ipsilateral to epileptogenesis than contralateral using multiple methods of analysis. PMID- 15788546 TI - Focal dystonia associated with pain. PMID- 15788547 TI - Is the disruption of the blood-brain barrier a prerequisite for cellular infiltration in autoimmune encephalitis? PMID- 15788548 TI - Intractable cluster headache and therapeutic stimulation of the hypothalamus: pathophysiological and management insights from a rare experiment. PMID- 15788549 TI - Impoverishment of spontaneous language and the prediction of Alzheimer's disease. PMID- 15788552 TI - Glutaric aciduria type 1: a clinician's view of progress. PMID- 15788553 TI - Hope, meaning, and growth following the September 11, 2001, terrorist attacks. AB - Positive psychologists found the increase of seven character strengths that encompass the so-called theological virtues, including hope and spirituality, in Americans after the September 11, 2001, attacks. Little is known about how they may affect post-September 11, 2001, mental health. Using multivariate analysis, this study investigated the relationship of hope and spiritual meaning with depression and anxiety in a sample of 457 students 3 months after September 11, 2001. Both characters contributed to lower levels of symptoms. In qualitative analysis, of 313 answers to an open-ended question regarding personal change, four categories emerged. The first three were consonant with other studies on posttraumatic growth (PTG), including changes in the self or behavior, relationships, and worldviews. The fourth category unique to September 11, 2001, was changes in political views. These findings offer further credence to the study of positive aspects resulting from violence-related trauma and highlight the needs for addressing the nature of traumatic events and PTG. PMID- 15788554 TI - Psychopathy, treatment change, and recidivism in high-risk, high-need sexual offenders. AB - The present study investigated 154 consecutive admissions to the Regional Treatment Center (Ontario) Sex Offender Treatment Program with reference to psychopathy and outcome. Ratings of treatment behavior, as well as clinical judgments as to whether risk was reduced, were coded based on treatment reports. With reference to Psychopathy Checklist-revised (PCL-R) scores, survival analyses indicated that high scorers recidivated at significantly higher rates than low scorers. However, offenders who received high PCL-R scores and lower scores on measures of treatment behavior recidivated at the same rate as low scorers on the PCL-R. Furthermore, among high PCL-R offenders, those rated as lower risk at post treatment in fact reoffended at a lower rate than those whose risk was rated as unchanged, although this difference failed to reach significance. Findings are discussed in light of the clinical and research literature. PMID- 15788555 TI - Acculturation and reported intimate partner violence among Latinas in Los Angeles. AB - This study sought to understand the relationship between acculturation and reporting intimate partner violence (IPV) among Latinas. A cross-sectional interviewer-administered survey was conducted at public health care clinics throughout Los Angeles County. Logistic regression was used to estimate the effect of acculturation on reporting IPV. An increasing trend of reporting IPV was observed among Latinas who were more acculturated (chi-square = 41.02, p = .0006). Highly acculturated Latinas were more likely to report IPV compared with least acculturated Latinas (prevalence odds ration = 2.18, 95% confidence level = 0.98, 4.89) and moderately acculturated Latinas were more likely to report IPV compared with least acculturated Latinas (prevalence odds ration = 1.29, 95% confidence level = 0.69, 2.43). Culturally competent IPV prevention programs may be the key to significantly reducing the number of women exposed to this serious public health problem. PMID- 15788556 TI - Barriers to services for rural and urban survivors of rape. AB - A significant proportion of survivors of rape do not utilize formal services to cope with the aftermath of rape. Understanding victimization experiences in environments that differ on resources, such as rural versus urban areas, may be an important dimension to consider in understanding barriers. Thirty women (18 rural and 12 urban) were recruited from rape crisis centers to participate in focus groups. Study results suggest that (a) survivors of rape experience many barriers to service utilization, (b) there were some differences in barriers to service utilization that were mentioned only in rural areas and some that were mentioned only in urban areas that may suggest that community context is important to consider in understanding barriers to service use, and (c) barriers to health and mental health services overlap with barriers to criminal justice system services. PMID- 15788557 TI - The role of insecure attachment and gender role stress in predicting controlling behaviors in men who batter. AB - The authors hypothesized that masculine gender role stress would mediate the relationship between insecure attachment and controlling behaviors in a sample of men who batter. To examine this hypothesis,143 men who were court mandated to attend a batterers' intervention program in a northeastern state completed measures including the Controlling Behavior Index, the Gender Role Stress Scale, the Relationship Questionnaire, and the Marlowe-Crowne Social Desirability Scale. After controlling statistically for social desirability, results indicated that both fearful attachment and gender role stress significantly predicted controlling behaviors, with gender role stress partially mediating the relationship between fearful attachment and controlling behaviors. The discussion focuses on the importance of understanding partner abuse through a gendered context. PMID- 15788558 TI - Weekly community interviews with high-risk participants: operational issues. AB - To address several key questions in social science research, repeated interviews of individuals drawn from difficult populations are required. This article describes an approach for addressing the challenges associated with longitudinal interview studies, including locating research participants, obtaining reliable and valid interview data over time, and retaining participants across the course of the study. We applied this approach to conduct a study designed to identify changeable risk factors for violence among high-risk people with mental illness. To successfully conduct weekly interviews of these individuals in the community across a 6-month period, we developed a flexible and personalized interview format; carefully selected, trained, and supervised staff; and developed incentives to maximize participant retention. Each of these three steps is discussed as a guide for future longitudinal studies that involve interviewing difficult populations. PMID- 15788559 TI - Feds to stockpile antivirals as pandemic "speed bump". PMID- 15788560 TI - Roles of polymerization dynamics, opposed motors, and a tensile element in governing the length of Xenopus extract meiotic spindles. AB - Metaphase spindles assemble to a steady state in length by mechanisms that involve microtubule dynamics and motor proteins, but they are incompletely understood. We found that Xenopus extract spindles recapitulate the length of egg meiosis II spindles, by using mechanisms intrinsic to the spindle. To probe these mechanisms, we perturbed microtubule polymerization dynamics and opposed motor proteins and measured effects on spindle morphology and dynamics. Microtubules were stabilized by hexylene glycol and inhibition of the catastrophe factor mitotic centromere-associated kinesin (MCAK) (a kinesin 13, previously called XKCM) and destabilized by depolymerizing drugs. The opposed motors Eg5 and dynein were inhibited separately and together. Our results are consistent with important roles for polymerization dynamics in regulating spindle length, and for opposed motors in regulating the relative stability of bipolar versus monopolar organization. The response to microtubule destabilization suggests that an unidentified tensile element acts in parallel with these conventional factors, generating spindle shortening force. PMID- 15788561 TI - Genome-wide analysis of the functions of a conserved surface on the corepressor Tup1. AB - The general transcriptional repressor Tup1 is responsible for the regulation of a large, diverse set of genes in Saccharomyces cerevisiae, and functional homologues of Tup1 have been identified in many metazoans. The crystal structure for the C-terminal portion of Tup1 has been solved and, when sequences of Tup1 homologues from fungi and metazoans were compared, a highly conserved surface was revealed. In this article, we analyze five point mutations that lie on this conserved surface. A statistical analysis of expression microarrays demonstrates that the mutant alleles are deficient in the repression of different subsets of Tup1-regulated genes. We were able to rank the mutant alleles of TUP1 based on the severity of their repression defects measured both by the number of genes derepressed and by the magnitude of that derepression. For one particular class of genes, the mutations on the conserved surface disrupted recruitment of Tup1 to the repressed promoters. However, for the majority of the genes derepressed by the Tup1 point mutants, recruitment of Tup1 to the regulated promoters is largely unaffected. These mutations affect the mechanism of repression subsequent to recruitment of the complex and likely represent a disruption of a mechanism that is conserved in fungi and metazoans. This work demonstrates that the evolutionarily conserved surface of Tup1 interacts with two separate types of proteins-sequence-specific DNA-binding proteins responsible for recruiting Tup1 to promoters as well as components that are likely to function in a conserved repression mechanism. PMID- 15788562 TI - Structural and functional characterization of noncoding repetitive RNAs transcribed in stressed human cells. AB - Thermal and chemical stresses induce the formation in human cells of novel and transient nuclear structures called nuclear stress bodies (nSBs). These contain heat shock factor 1 (HSF-1) and a specific subset of pre-mRNA processing factors. Nuclear stress bodies are assembled on specific pericentromeric heterochromatic domains containing satellite III (SatIII) DNA. In response to stress, these domains change their epigenetic status from heterochromatin to euchromatin and are transcribed in poly-adenylated RNAs that remain associated with nSBs. In this article, we describe the cloning, sequencing, and functional characterization of these transcripts. They are composed of SatIII repeats and originate from the transcription of multiple sites within the SatIII arrays. Interestingly, the level of SatIII RNAs can be down-regulated both by antisense oligonucleotides and small interfering RNAs (siRNA). Knockdown of SatIII RNA by siRNAs requires the activity of Argonaute 2, a component of the RNA-induced silencing complex. Down regulation of satellite III RNAs significantly affects the recruitment of RNA processing factors to nSBs without altering the association of HSF-1 with these structures nor the presence of acetylated histones within nSBs. Thus, satellite III RNAs have a major role in the formation of nSBs. PMID- 15788563 TI - SUMO represses transcriptional activity of the Drosophila SoxNeuro and human Sox3 central nervous system-specific transcription factors. AB - Sry high mobility group (HMG) box (Sox) transcription factors are involved in the development of central nervous system (CNS) in all metazoans. Little is known on the molecular mechanisms that regulate their transcriptional activity. Covalent posttranslational modification by small ubiquitin-like modifier (SUMO) regulates several nuclear events, including the transcriptional activity of transcription factors. Here, we demonstrate that SoxNeuro, an HMG box-containing transcription factor involved in neuroblast formation in Drosophila, is a substrate for SUMO modification. SUMOylation assays in HeLa cells and Drosophila S2 cells reveal that lysine 439 is the major SUMO acceptor site. The sequence in SoxNeuro targeted for SUMOylation, IKSE, is part of a small inhibitory domain, able to repress in cis the activity of two adjacent transcriptional activation domains. Our data show that SUMO modification represses SoxNeuro transcriptional activity in transfected cells. Overexpression in Drosophila embryos of a SoxN form that cannot be targeted for SUMOylation strongly impairs the development of the CNS, suggesting that SUMO modification of SoxN is crucial for regulating its activity in vivo. Finally, we present evidence that SUMO modification of group B1 Sox factors was conserved during evolution, because Sox3, the human counterpart of SoxN, is also negatively regulated through SUMO modification. PMID- 15788564 TI - Drosophila activating transcription factor-2 is involved in stress response via activation by p38, but not c-Jun NH(2)-terminal kinase. AB - Activating transcription factor (ATF)-2 is a member of the ATF/cAMP response element-binding protein family of transcription factors, and its trans-activating capacity is enhanced by stress-activated protein kinases such as c-Jun NH(2) terminal kinase (JNK) and p38. However, little is known about the in vivo roles played by ATF-2. Here, we identified the Drosophila homologue of ATF-2 (dATF-2) consisting of 381 amino acids. In response to UV irradiation and osmotic stress, Drosophila p38 (dp38), but not JNK, phosphorylates dATF-2 and enhances dATF-2 dependent transcription. Consistent with this, injection of dATF-2 double stranded RNA (dsRNA) into embryos did not induce the dorsal closure defects that are commonly observed in the Drosophila JNK mutant. Furthermore, expression of the dominant-negative dp38 enhanced the aberrant wing phenotype caused by expression of a dominant-negative dATF-2. Similar genetic interactions between dATF-2 and the dMEKK1-dp38 signaling pathway also were observed in the osmotic stress-induced lethality of embryos. Loss of dATF-2 in Drosophila S2 cells by using dsRNA abrogated the induction of 40% of the osmotic stress-induced genes, including multiple immune response-related genes. This indicates that dATF-2 is a major transcriptional factor in stress-induced transcription. Thus, dATF-2 is critical for the p38-mediated stress response. PMID- 15788566 TI - Differential subnuclear localization and replication timing of histone H3 lysine 9 methylation states. AB - Mono-, di-, and trimethylation of specific histone residues adds an additional level of complexity to the range of histone modifications that may contribute to a histone code. However, it has not been clear whether different methylated states reside stably at different chromatin sites or whether they represent dynamic intermediates at the same chromatin sites. Here, we have used recently developed antibodies that are highly specific for mono-, di-, and trimethylated lysine 9 of histone H3 (MeK9H3) to examine the subnuclear localization and replication timing of chromatin containing these epigenetic marks in mammalian cells. Me1K9H3 was largely restricted to early replicating, small punctate domains in the nuclear interior. Me2K9H3 was the predominant MeK9 epitope at the nuclear and nucleolar periphery and colocalized with sites of DNA synthesis primarily in mid-S phase. Me3K9H3 decorated late-replicating pericentric heterochromatin in mouse cells and sites of DAPI-dense intranuclear heterochromatin in human and hamster cells that replicated throughout S phase. Disruption of the Suv39h1,2 or G9a methyltransferases in murine embryonic stem cells resulted in a redistribution of methyl epitopes, but did not alter the overall spatiotemporal replication program. These results demonstrate that mono-, di-, and trimethylated states of K9H3 largely occupy distinct chromosome domains. PMID- 15788565 TI - Myosin Va transports dense core secretory vesicles in pancreatic MIN6 beta-cells. AB - The role of unconventional myosins in neuroendocrine cells is not fully understood, with involvement suggested in the movement of both secretory vesicles and mitochondria. Here, we demonstrate colocalization of myosin Va (MyoVa) with insulin in pancreatic beta-cells and show that MyoVa copurifies with insulin in density gradients and with the vesicle marker phogrin-enhanced green fluorescent protein upon fluorescence-activated sorting of vesicles. By contrast, MyoVa immunoreactivity was poorly colocalized with mitochondrial or other markers. Demonstrating an important role for MyoVa in the recruitment of secretory vesicles to the cell surface, a reduction of MyoVa protein levels achieved by RNA interference caused a significant decrease in glucose- or depolarization stimulated insulin secretion. Similarly, expression of the dominant-negative acting globular tail domain of MyoVa decreased by approximately 50% the number of vesicles docked at the plasma membrane and by 87% the number of depolarization stimulated exocytotic events detected by total internal reflection fluorescence microscopy. We conclude that MyoVa-driven movements of vesicles along the cortical actin network are essential for the terminal stages of regulated exocytosis in beta-cells. PMID- 15788567 TI - The Xenopus TACC homologue, maskin, functions in mitotic spindle assembly. AB - Maskin is the Xenopus homolog of the transforming acidic coiled coil (TACC) family of microtubule and centrosome-interacting proteins. Members of this family share a approximately 200 amino acid coiled coil motif at their C-termini, but have only limited homology outside of this domain. In all species examined thus far, perturbations of TACC proteins lead to disruptions of cell cycle progression and/or embryonic lethality. In Drosophila, Caenorhabditis elegans, and humans, these disruptions have been attributed to mitotic spindle assembly defects, and the TACC proteins in these organisms are thought to function as structural components of the spindle. In contrast, cell division failure in early Xenopus embryo blastomeres has been attributed to a role of maskin in regulating the translation of, among others, cyclin B1 mRNA. In this study, we show that maskin, like other TACC proteins, plays a direct role in mitotic spindle assembly in Xenopus egg extracts and that this role is independent of cyclin B. Maskin immunodepletion and add-back experiments demonstrate that maskin, or a maskin associated activity, is required for two distinct steps during spindle assembly in Xenopus egg extracts that can be distinguished by their response to "rescue" experiments. Defects in the "early" step, manifested by greatly reduced aster size during early time points in maskin-depleted extracts, can be rescued by readdition of purified full-length maskin. Moreover, defects in this step can also be rescued by addition of only the TACC-domain of maskin. In contrast, defects in the "late" step during spindle assembly, manifested by abnormal spindles at later time points, cannot be rescued by readdition of maskin. We show that maskin interacts with a number of proteins in egg extracts, including XMAP215, a known modulator of microtubule dynamics, and CPEB, a protein that is involved in translational regulation of important cell cycle regulators. Maskin depletion from egg extracts results in compromised microtubule asters and spindles and the mislocalization of XMAP215, but CPEB localization is unaffected. Together, these data suggest that in addition to its previously reported role as a translational regulator, maskin is also important for mitotic spindle assembly. PMID- 15788568 TI - The prion protein and its paralogue Doppel affect calcium signaling in Chinese hamster ovary cells. AB - The function of the prion protein (PrP(c)), implicated in transmissible spongiform encephalopathies (TSEs), is largely unknown. We examined the possible influence of PrP(c) on Ca(2+) homeostasis, by analyzing local Ca(2+) fluctuations in cells transfected with PrP(c) and Ca(2+)-sensitive aequorin chimeras targeted to defined subcellular compartments. In agonist-stimulated cells, the presence of PrP(c) sharply increases the Ca(2+) concentration of subplasma membrane Ca(2+) domains, a feature that may explain the impairment of Ca(2+)-dependent neuronal excitability observed in TSEs. PrP(c) also limits Ca(2+) release from the endoplasmic reticulum and Ca(2+) uptake by mitochondria, thus rendering unlikely the triggering of cell death pathways. Instead, cells expressing Doppel, a PrP(c) paralogue, display opposite effects, which, however, are abolished by the coexpression of PrP(c). These findings are consistent with the functional interplay and antagonistic role attributed to the proteins, whereby PrP(c) protects, and Doppel sensitizes, cells toward stress conditions. PMID- 15788571 TI - Radiology's role in covering all the bases. PMID- 15788569 TI - The PCH family member MAYP/PSTPIP2 directly regulates F-actin bundling and enhances filopodia formation and motility in macrophages. AB - Macrophage actin-associated tyrosine phosphorylated protein (MAYP) belongs to the Pombe Cdc15 homology (PCH) family of proteins involved in the regulation of actin based functions including cell adhesion and motility. In mouse macrophages, MAYP is tyrosine phosphorylated after activation of the colony-stimulating factor-1 receptor (CSF-1R), which also induces actin reorganization, membrane ruffling, cell spreading, polarization, and migration. Because MAYP associates with F actin, we investigated the function of MAYP in regulating actin organization in macrophages. Overexpression of MAYP decreased CSF-1-induced membrane ruffling and increased filopodia formation, motility and CSF-1-mediated chemotaxis. The opposite phenotype was observed with reduced expression of MAYP, indicating that MAYP is a negative regulator of CSF-1-induced membrane ruffling and positively regulates formation of filopodia and directional migration. Overexpression of MAYP led to a reduction in total macrophage F-actin content but was associated with increased actin bundling. Consistent with this, purified MAYP bundled F actin and regulated its turnover in vitro. In addition, MAYP colocalized with cortical and filopodial F-actin in vivo. Because filopodia are postulated to increase directional motility by acting as environmental sensors, the MAYP stimulated increase in directional movement may be at least partly explained by enhancement of filopodia formation. PMID- 15788572 TI - Statistical inference for continuous variables. PMID- 15788573 TI - Statistical inference for proportions. PMID- 15788574 TI - Seven leadership fallacies and how to correct them. PMID- 15788570 TI - Glycoprotein gp130 of dictyostelium discoideum influences macropinocytosis and adhesion. AB - Glycoprotein gp130, found on the plasma membrane of Dictyostelium discoideum amoebae, was postulated previously to play a role in phagocytosis. The gene for gp130 was cloned and when translated, yielded a 768 amino acid preproprotein of 85.3 kDa. It had nearly 40% similarity to the 138 kDa family of glycoproteins implicated in sexual cell fusion during macrocyst formation in D. discoideum. The difference between the calculated size and observed M(r) of 130 kDa on protein gels likely was due to N-glycosylation that was confirmed by lectin blots. Consistent with its surface-exposure, an antibody raised against recombinant protein stained the plasma membrane of D. discoideum amoebae. Gp130 and its transcripts were high during axenic growth of cells, but relatively low during growth on bacteria. The gene for gp130 was disrupted and cell lines lacking the glycoprotein were efficient phagocytes, indicating that gp130 was dispensable for phagocytosis. Gp130-null cells were similar in size to parent DH1 cells, had enhanced macropinocytosis and grew faster to higher densities. They also exhibited weaker cell-substrate adhesion but displayed greater cell-cell cohesion. Collectively, the data indicated that gp130 influenced macropinocytosis and played a role in adhesion during vegetative growth. PMID- 15788575 TI - Ferucarbotran-enhanced MRI versus triple-phase MDCT for the preoperative detection of hepatocellular carcinoma. AB - OBJECTIVE: We compared ferucarbotran-enhanced MRI with triple-phase MDCT for the preoperative detection of hepatocellular carcinoma. SUBJECTS AND METHODS: Seventy three consecutive patients with 121 hepatocellular carcinomas underwent ferucarbotran-enhanced MRI, including a dynamic study, and triple-phase MDCT before hepatic resection. The diagnosis of hepatocellular carcinoma was confirmed in all patients by means of pathologic examination after surgical resection. Three experienced radiologists independently reviewed the MR and CT images on a segment-by-segment basis. The accuracy of these techniques for the detection of hepatocellular carcinoma was assessed by conducting a receiver operating characteristic (ROC) analysis of the observations of 88 resected hepatic segments with at least one hepatocellular carcinoma each and 121 resected hepatic segments without hepatocellular carcinoma. RESULTS: The mean values of the area under the ROC curve (A(z)) for ferucarbotran-enhanced MRI and triple-phase MDCT for all observers were 0.947 and 0.949, respectively; the difference between these two values was not statistically significant (p = 0.799). The mean sensitivities of MRI and triple-phase MDCT were 90.2% and 91.3%, respectively, and their mean specificities were 97.0% and 95.3%, respectively. The differences in the mean sensitivities and specificities of these two imaging techniques were not statistically significant (p > 0.05 in each case). CONCLUSION: Ferucarbotran enhanced MRI seems to be as accurate as triple-phase MDCT for the preoperative detection of hepatocellular carcinoma. PMID- 15788576 TI - Value of contrast-enhanced sonography for the characterization of focal hepatic lesions in patients with diffuse liver disease: receiver operating characteristic analysis. AB - OBJECTIVE: Our aim was to assess the diagnostic performance of contrast-enhanced agent detection sonographic imaging to characterize focal hepatic lesions in patients with diffuse liver disease in comparison with baseline sonographic images and to determine whether agent detection imaging can reduce the necessity of further diagnostic workup for lesion characterization. MATERIALS AND METHODS: Contrast-enhanced sonography using 4 g of Levovist at a concentration of 300 mg/mL was performed on 75 focal hepatic lesions in 75 patients with diffuse liver disease. Interval reviews for both baseline without and with contrast-enhanced sonography were performed independently by two radiologists. They were requested to determine the malignity of focal hepatic lesions using a 5-point confidence level and to record the specific diagnoses and the necessity for further imaging for lesion characterization. Radiologists' performances for lesion differentiation using baseline and contrast-enhanced sonography were evaluated using receiver operating characteristic (ROC) analysis. Interobserver agreement was also analyzed. RESULTS: When contrast-enhanced sonography was used, ROC analysis revealed a significant improvement for both reviewers (area under the receiver operating characteristic curve [A(z)] = 0.753 and 0.830 and 0.971 and 0.974 at baseline sonography and contrast-enhanced sonography, respectively; p < 0.002) for differentiating malignant and benign focal liver lesions. Contrast enhanced sonography also improved specificity from 12% to 91% for reviewer 1 and from 26% to 85% for reviewer 2 compared with baseline sonography. Furthermore, excellent interobserver agreement was achieved for contrast-enhanced sonography (weighted kappa = 0.919), whereas only good agreement was achieved for baseline sonography (weighted kappa = 0.656). A better result for specific diagnosis was obtained by contrast-enhanced sonography (79% and 75%) than by baseline sonography (37% and 48%, p < 0.05). Contrast-enhanced sonography (72% and 63%) outperformed baseline sonography (35% and 28%, p < 0.05) as a confirmatory imaging technique. CONCLUSION: Contrast-enhanced agent detection sonography can be used to characterize focal hepatic lesions in patients with diffuse liver disease reliably and with a higher diagnostic confidence than baseline sonography. Furthermore, contrast-enhanced sonography reduced the need for further diagnostic workups for focal hepatic lesion characterization. PMID- 15788577 TI - Focal eosinophilic necrosis versus metastasis in the liver: the usefulness of two phase dynamic CT. AB - OBJECTIVE: The purpose of our study was to evaluate the usefulness of dual-phase dynamic CT in the differentiation of focal eosinophilic necrosis of the liver and metastasis. CONCLUSION: Undetected isoattenuating lesions on arterial phase images that have an indistinct margin, a nonspherical shape, and a homogenous enhancement pattern on portal venous phase images suggest focal eosinophilic necrosis rather than metastasis. Two-phase dynamic CT was found to be useful at differentiating focal eosinophilic necrosis from metastasis. PMID- 15788578 TI - Radiation injury to the liver after intensity-modulated radiation therapy in patients with mesothelioma: an unusual CT appearance. AB - OBJECTIVE: We sought to report the unusual distribution of radiation-induced injury to the liver in patients with mesothelioma after extrapleural pneumonectomy and intensity-modulated radiation therapy (IMRT). CONCLUSION: Abnormal hepatic enhancement after extrapleural pneumonectomy and IMRT is common in patients with mesothelioma. Knowledge of the early occurrence and typical location and appearance of IMRT-induced injury can be useful in preventing misinterpretation as metastatic disease or recurrent tumor. PMID- 15788579 TI - Evaluation of liver metastases after radiofrequency ablation: utility of 18F-FDG PET and PET/CT. AB - OBJECTIVE: Our objective was to review the CT appearance of liver metastases after radiofrequency ablation and to describe the imaging findings of and utility of (18)F-FDG PET and PET/CT in assessing tumor recurrence after ablation. CONCLUSION: (18)F-FDG PET and PET/CT can provide added diagnostic information compared with conventional imaging in patients after radiofrequency ablation of liver metastases and can be useful in guiding repeat ablation procedures. PMID- 15788580 TI - Fat-containing lesions of the liver: cross-sectional imaging findings with emphasis on MRI. AB - OBJECTIVE: The purpose of this pictorial essay is to identify different types of liver lesions that contain fat. Cross-sectional imaging findings of fat- or lipid containing lesions can help in characterizing focal liver lesions. We searched our archive retrospectively and reviewed the literature for fat-containing liver lesions and identified 16 different types. CONCLUSION: These lesions can contain macroscopic fat (i.e., angiomyolipoma, lipoma, liposarcoma, hydatid cyst, lipopeliosis, adrenal rest tumor, pseudolipoma, hepatic teratoma, pericaval fat, extramedullary hematopoiesis, and metastases) or intracellular lipid (i.e., focal steatosis, adenoma, focal nodular hyperplasia, regenerative nodules, and hepatocellular carcinoma). CT, MRI, and sonographic findings of these lesions can help in characterization by allowing specific diagnosis or narrowing the differential diagnosis of liver lesions. PMID- 15788581 TI - Value of the single-phase technique in MDCT assessment of pancreatic tumors. AB - OBJECTIVE: The purpose of our study was to determine the diagnostic value of single-phase MDCT in patients with suspected pancreatic carcinoma. SUBJECTS AND METHODS: Seventy-one patients (41 men, 30 women; mean age, 63 years; range, 29-80 years) with suspected pancreatic tumor underwent MDCT. Scanning was performed on an MDCT scanner with 0.5-sec gantry rotation and acquisition of 4 slices per rotation. Unenhanced scanning was followed by one set of scanning in the caudocranial direction from the inferior hepatic margin to the diaphragm with a scanning delay of 60 sec after the IV injection of 150 mL of contrast material delivered at 3 mL/sec. Two reviewers independently scored images in a blinded fashion for the presence of tumor and assessment of resectability. Receiver operating characteristic analysis was performed. RESULTS: A final histopathologic diagnosis derived from surgical findings was obtained in 42 patients; in the remaining 29 patients, percutaneous fine-needle aspiration biopsy coupled with a 1-year clinical follow-up to determine development of local, regional or distant neoplasm served as gold standard proof of diagnosis. Final diagnosis was pancreatic cancer in 40 patients (27 ductal adenocarcinoma, nine mucinous cystoadenocarcinoma, two neuroendocrine tumors, one lymphoma, and one papillary cystoadenocarcinoma) and chronic pancreatitis in 31. The mean tumor size was 2.4 cm (range, 4-1 cm). Values for the area under the curve (A(z)) for the assessment of tumor detection were 0.97 for reviewer 1 and 0.96 for reviewer 2 (p = not significant). A(z) values for tumor resectability were 0.90 for reviewer 1 and 0.90 for reviewer 2 (p = not significant). No statistically significant differences were observed between superior mesenteric artery and vein opacification with the hepatic parenchyma enhanced at a time closer to the peak hepatic enhancement, optimizing the detection of hepatic lesions. CONCLUSION: Thin-section single-phase MDCT is an accurate technique for the diagnosis and assessment of resectability in patients with a suspected pancreatic neoplasm. This technique provides optimal tumor-to-pancreas contrast and maximal pancreatic parenchymal and peripancreatic vascular enhancement. It allows visualization of the entire liver and the whole upper abdomen during the portal phase for accurate identification of liver metastases and peritoneal seeding. PMID- 15788582 TI - Surgery for chronic pancreatitis: cross-sectional imaging of postoperative anatomy and complications. AB - OBJECTIVE: Surgery is increasingly undertaken for intractable chronic pancreatitis. We evaluated the postsurgical anatomy and complications of surgical options including Whipple, Puestow, Frey's, and Beger's procedures. CONCLUSIONS: Knowledge of postsurgical anatomy is important to avoid misdiagnosing expected anatomy as complications on CT examinations. It is important to carefully assess the upper abdominal arteries to detect subtle but potentially lethal complications. PMID- 15788583 TI - MRI features of mesenteric desmoid tumors in familial adenomatous polyposis. PMID- 15788584 TI - Helical CT evaluation of acute right lower quadrant pain: part I, common mimics of appendicitis. AB - OBJECTIVE: The purpose of our pictorial essay is to present common mimics of appendicitis as noted on helical CT in patients with right lower quadrant pain and to highlight the features that provide clues to the diagnosis. CONCLUSION: Recognition of the findings of common diseases that simulate acute appendicitis on helical CT, along with features that help to differentiate these entities from appendicitis, is important in establishing a correct diagnosis and in guiding appropriate therapy. PMID- 15788585 TI - Helical CT evaluation of acute right lower quadrant pain: part II, uncommon mimics of appendicitis. AB - OBJECTIVE: The purpose of our pictorial essay is to highlight the helical CT features of uncommon mimics of appendicitis and to provide clues to differentiate them from appendicitis. CONCLUSION: Uncommon mimics of appendicitis create a diagnostic challenge in patients with right lower quadrant pain. Recognition of the helical CT features of uncommon mimics of appendicitis is important in clinical management and avoiding unnecessary laparotomy. PMID- 15788586 TI - Contrast-enhanced sonography of the spleen. AB - OBJECTIVE: Combined use of low-mechanical-index technologies and non-air-filled contrast media allows real-time sonographic assessment of the spleen. This pictorial essay focuses on several aspects of contrast-enhanced sonography of the spleen, including examination technique, clinical indications, normal findings obtained through all vascular phases, abnormal findings (splenomegaly, infarction, abscess, benign tumors, lymphoma, metastasis, injuries, and accessory spleen), limitations, and pitfalls. CONCLUSION: Real-time, contrast-enhanced sonography is a novel technique allowing depiction of a wide range of splenic abnormalities. PMID- 15788587 TI - Detection of combined hepatocellular and cholangiocarcinomas on enhanced CT: comparison with histologic findings. AB - OBJECTIVE: The purpose of this study was to evaluate the performance of enhanced CT in making a diagnosis of combined hepatocellular and cholangiocarcinomas (HCC CCs) by comparing CT findings with histologic findings. CONCLUSION: One third (nine of 27 cases) of the combined HCC-CCs were correctly diagnosed on enhanced CT by detailed analysis of the enhancing pattern around or within the mass. Various factors such as an atypical enhancing pattern, the size of each component, and the presence of a mass composed of intermediate tumor cells-that is, cells with intermediate characteristics between HCC and CC-were found to be the causes of misdiagnosis of combined HCC-CC on enhanced CT. PMID- 15788588 TI - Imaging and findings of lipomas of the gastrointestinal tract. PMID- 15788589 TI - Persistent foreign body reaction around inguinal mesh prostheses: a potential pitfall of FDG PET. AB - OBJECTIVE: FDG PET has been recognized as an efficient imaging technique for the treatment of oncology patients. However, false-positive results can occur. The purpose of this study is to describe three oncology patients with persistent FDG up-take around inguinal mesh prostheses that occurred up to 10 years after the surgical repair of inguinal hernias and led to false-positive results. CONCLUSION: Remote mesh prostheses can induce FDG uptake because of persistent foreign body reaction. Consequently, each time an unexpected pelvic focus is noticed on FDG PET, the medical history of patients should be carefully reviewed to avoid false-positive results. PMID- 15788590 TI - Gadolinium-enhanced CT angiography of endovascular stent-grafts. PMID- 15788591 TI - Polyp detection with MDCT: a phantom-based evaluation of the impact of dose and spatial resolution. AB - OBJECTIVE: The objective of our study was to evaluate the impact of dose and spatial resolution on the detection of colonic polyps using a 4-MDCT scanner. MATERIALS AND METHODS: Twenty-four latex phantoms that simulate the large bowel and contain artificial polyps of different sizes and shapes were constructed. The polyps were divided into three size groups (diameter, 0-2, 2-5, and 5-10 mm) and were classified into four shape groups: pedunculated; broad-based; ulcerated or depressed; and sessile or flat. The colon phantoms were submerged in a water tank and scanned on a 4-MDCT scanner using 12 protocols with various settings of slice thickness, pitch, and tube current. The images were independently evaluated by three radiologists using axial 2D multiplanar reconstruction images and a 3D surface-rendering technique (fly-through). RESULTS: At a constant dose (i.e., dose-length product [DLP]), the polyp detection rate increased with increasing axial spatial resolution. For the standard protocol (2.50-mm slice thickness, 1.5 pitch), the detection rate for all polyp sizes decreased from approximately 70% at 100 mA to 55% at 40 mA. Between a 60- and 100-mA tube current, the detection rate for the largest polyps (> 5 mm) was almost constant, close to 90%. CONCLUSION: The detection of polyps in the large bowel using a standard protocol can be improved without dose penalty by increasing the axial spatial resolution of the image acquisition and adjusting the tube current setting. If the analysis can be restricted to polyps larger than 5 mm, the dose can be substantially reduced without compromising the detection rate. PMID- 15788592 TI - Stercoral colitis leading to fatal peritonitis: CT findings. AB - OBJECTIVE: Stercoral colitis is an inflammatory process involving the colonic wall related to fecal impaction. Our purpose was to describe the imaging findings of stercoral colitis and ulceration and to emphasize the potential serious clinical implications of the condition. CONCLUSION: Fecal impaction may lead to ischemic pressure necrosis and subsequent colonic perforation. In the appropriate clinical setting, the imaging findings that should prompt the radiologist to consider this diagnosis are the presence of fecal impaction, focal colonic wall thickening, and adjacent stranding of the fat. If the fecal impaction is not promptly relieved, the condition can lead to colonic perforation, peritonitis, and patient demise. PMID- 15788593 TI - Vascular virtual endoluminal visualization of invasive colorectal cancer on MDCT colonography. AB - OBJECTIVE: The purpose of this study was to assess the utility of vascular views for visualization of invasive colorectal cancers on contrast-enhanced MDCT colonography. CONCLUSION: By means of Hounsfield-transparency settings, we obtained virtual endoluminal images that show vascular structures and delineate invasive cancers of the colorectal wall, and we call these images "vascular views." Using this technique for contrast-enhanced MDCT colonography, we found that the increase in flow and pooling of blood related to angiogenesis of cancerous lesions is easy to identify and that this finding is useful in the detection of invasive colorectal cancers. PMID- 15788594 TI - Value of FDG PET in the assessment of patients with multiple myeloma. AB - OBJECTIVE: Our objective was to evaluate if whole-body PET with FDG is able to detect bone marrow involvement in patients with multiple myeloma and to assess its appearance and distribution pattern. MATERIALS AND METHODS: Seventeen whole body FDG PET scans were performed in 13 patients with multiple myeloma. Four patients were referred for evaluation of extent of disease pretherapy and nine patients were referred for assessment of therapy response (chemotherapy, radiation therapy, bone marrow transplant). FDG PET images were evaluated for distribution and uptake pattern. Standardized uptake values were obtained to quantify FDG uptake. Results of other imaging examinations (MRI, CT, radiography), laboratory data, biopsies, and the clinical course were used for verification of detected lesions. RESULTS: FDG PET was able to detect medullary involvement of multiple myeloma. There were two false-negative results. In one patient, the radiographic skeletal survey showed subcentimeter lytic lesions within the ribs that were not detected on FDG PET and in the other patient, a lytic lesion detected on radiographs showed only mildly increased FDG uptake that was not identified prospectively. There was one false-positive FDG PET result in a patient who had undergone radiation therapy 3 weeks before PET. FDG PET was helpful in differentiating between posttherapeutic changes and residual/recurrent tumor and in assessing response to therapy. FDG PET resulted in upstaging of disease in four patients, which influenced subsequent management and prognosis. Sensitivity of FDG PET in detecting myelomatous involvement was 85% and specificity was 92%. CONCLUSION: FDG PET is able to detect bone marrow involvement in patients with multiple myeloma. FDG PET is useful in assessing extent of disease at time of initial diagnosis, contributing to staging that is more accurate. FDG PET is also useful for evaluating therapy response. PMID- 15788595 TI - Sonographic evaluation of injuries to the pectoralis muscles. AB - OBJECTIVE: Our aim was to show the application of high-resolution sonography for grading and defining athletic and nonathletic injuries to the pectoralis muscles. CONCLUSION: Sonography provides dynamic high-resolution imaging for the diagnosis and grading of injuries to the pectoralis muscles. PMID- 15788596 TI - Cystic lesions in the posterosuperior portion of the humeral head on MR arthrography: correlations with gross and histologic findings in cadavers. AB - OBJECTIVE: The purpose of this study was to describe the appearance of cysts in the posterosuperior portion of the humeral head on MR arthrography and to correlate the MRI findings with the gross and histologic findings in cadavers. CONCLUSION: Cysts in the posterosuperior portion of the humeral head (the bare area) were located in the lateral humeral head just posterior to the greater tuberosity. These cysts were lined with collagen connective tissue and were connected to the joint spaces. We suggest that these pseudocysts in the posterosuperior portion of the humeral head may be a normal variant rather than being an abnormal change or vascular channel. PMID- 15788597 TI - Micro-CT arthrography: a pilot study for the ex vivo visualization of the rat knee joint. AB - OBJECTIVE: In our study, we evaluated the potential of micro-CT for the assessment of the rat knee joint using ex vivo micro-CT arthrography. The aims of the study were to introduce the technique of micro-CT arthrography and to visualize the normal anatomy of the rat knee. The secondary aims were the quantification of retropatellar cartilage thickness and the analysis of microstructural cancellous bone parameters within the tibial epiphysis. CONCLUSION: Micro-CT arthrography is a novel technique for the indirect visualization of the distinct features and structural analysis of the rat knee joint. This technique represents an additional imaging and analysis tool in small animal research. PMID- 15788598 TI - Electrochemical corrosion of metal implants. AB - OBJECTIVE: The objective of our study was to show the radiographic changes that result from electrochemical corrosion of implanted metal in the body. CONCLUSION: Corrosion of metal implants is not rare. Radiologists should become familiar with the changes this process produces. PMID- 15788599 TI - Tenosynovial chondromatosis of the ring finger. PMID- 15788600 TI - Effects of heart rate on motion artifacts of the aorta on non-ECG-assisted 0.5 sec thoracic MDCT. AB - OBJECTIVE: Our aim was to evaluate the effects of heart rate on aortic motion artifacts on 0.5-sec non-ECG-assisted thoracic MDCT. MATERIALS AND METHODS: A total of 124 non-ECG-assisted thoracic MDCT scans with satisfactory simultaneous ECG data were reviewed. Scans were grouped according to patient heart rates (beats per minute [bpm]: group A, 46-55; B, 56-65; C, 66-75; D, 76-85; E, 86-95; and F > 95). The groups were compared regarding the presence, locations, and spatial distributions of pulsation artifact, number of slices affected, maximum amplitude of pulsation, continuity of artifact, and the presence of superior vena cava (SVC) pseudoflaps. RESULTS: Of the 124 scans, 114 (91.9%) had aortic motion artifacts, with prevalence ranging from 85.3% (66-75 bpm) to 100% (65 bpm or less). Of the 114 motion artifacts, all affected the ascending aorta, 105 (92.1%) involved the left anterior and right posterior aspects of the aortic circumference, and 106 (93%) were associated with SVC pseudoflaps. Group B had significantly greater numbers of images with artifacts (p < 0.001-0.006), greater artifact amplitudes (p < 0.001-0.002), and a higher continuity trend for the artifacts (p = 0.003-0.194) than did the other five groups. CONCLUSION: Aortic motion artifacts are frequently seen on thoracic MDCT, especially in patients with heart rates of 65 bpm or less. The presence of a SVC pseudoflap is helpful for distinguishing artifacts from dissection. If aortic disease is suspected, then measures to reduce motion artifact, such as ECG-gating, should be considered. PMID- 15788601 TI - The effect of single-detector CT versus MDCT on clinical outcomes in patients with suspected acute pulmonary embolism and negative results on CT pulmonary angiography. AB - OBJECTIVE: We sought to compare the clinical outcomes of patients in whom pulmonary embolism (PE) has been ruled out with single-detector CT versus MDCT, given the improved visualization of subsegmental clots with the latter and the recent increase in use of CT for evaluation of PE. SUBJECTS AND METHODS: Two cohorts of patients undergoing CT for suspected PE with either single-detector CT (3-mm collimation and pitch of 1.7) or MDCT (2-mm collimation and pitch of 1) scanners were prospectively observed and compared using predefined criteria for evidence of subsequent thromboembolic disease during the 6 months after the acquisition of their initial scan. RESULTS: Ninety-eight patients were scanned using a single-detector CT scanner. Of these, none had evidence of subsequent PE or deep venous thrombosis (DVT), and six (6.1%) died of unrelated causes. Of the 100 patients scanned using an MDCT scanner, one (1.0%) had a subsequent nonfatal PE 2 months after the initial scanning, one (1.0%) had DVT 1 month after the initial scanning, and eight (8.0%) died of unrelated causes. No significant difference was found in either the probability of subsequent thromboembolic events (chi(2) = 0.3183, degrees of freedom [df] = 1, p = 1) or frequency of unrelated deaths (chi(2) = 0.2655, df = 1, p = 0.7829) between patients scanned using single-detector CT or MDCT protocols. CONCLUSION: Our results show that patients with suspected acute PE and negative CT results have acceptable clinical outcomes in the absence of anticoagulation treatment up to 6 months after acquisition of their initial scan. Furthermore, we found that the increased visualization of smaller, more peripheral arteries afforded by multislice technology did not affect clinical outcome. PMID- 15788602 TI - Anomalous brachiocephalic vein: CT, embryology, and clinical implications. AB - OBJECTIVE: CT patterns of anomalous brachiocephalic veins are presented with reconsideration of the structure's embryogenesis. CONCLUSION: With advancements in central line procedures and corrective cardiac surgery, and the widespread use of noninvasive imaging techniques, the clinical importance of identification of the anomalous brachiocephalic vein is shown. PMID- 15788603 TI - Aneurysm of an aberrant systemic artery to the lung. PMID- 15788604 TI - Intraaortic balloon pump location and aortic dissection. PMID- 15788605 TI - Hypothesis on the evolution of cavitary lesions in nontuberculous mycobacterial pulmonary infection: thin-section CT and histopathologic correlation. AB - OBJECTIVE: The objectives of our study were to evaluate the thin-section CT findings of the cavitary form of nontuberculous mycobacterial pulmonary infection and correlate these imaging findings with the histopathologic findings concerning the development of bronchiectasis and of centrilobular nodules and cavitary lesions. MATERIALS AND METHODS: We retrospectively reviewed thin-section CT scans (2.5-mm collimation, both axial and coronal reformation images) of 24 cases (male female ratio, 13:11; mean age, 61 years; age range, 43-82 years) of the cavitary form of culture-proven Mycobacterium avium-intracellulare complex pulmonary infection including two cases with lobectomy specimens. Any changes in CT findings detected on the follow-up CT scans that were available for seven patients (follow-up interval, 6-24 months; mean, 12 months) were also assessed. RESULTS: Thin-section CT findings were bronchiectasis (24/24 patients, 100%), a patent bronchus running into a cavitary lesion (the "feeding bronchus" appearance) (18/24, 75%), nodules less than 10 mm (17/24, 71%), centrilobular nodules (17/24, 71%), nodules of 10-30 mm (13/24, 54%), peribronchial nodules (8/24, 33%), lobular consolidation (6/24, 25%), bronchial wall thickening (4/24, 17%), and consolidation (2/24, 8%). Two lobectomy specimens showed large cavitary consolidations with the feeding bronchus appearance on pathologic specimens. In two patients, small peribronchial nodules had changed into cavitary nodules with the feeding bronchus appearance on follow-up CT, which represented inflamed focal cystic bronchiectasis. CONCLUSION: In the cavitary form of M. avium intracellulare complex pulmonary infection, the feeding bronchus appearance is another very frequent thin-section CT finding. This appearance may suggest that peribronchial nodules of M. avium-intracellulare complex infection evolve into inflamed focal cystic bronchiectasis manifesting as cavitary lesions. PMID- 15788606 TI - Peripheral pulmonary artery pseudoaneurysms and massive hemoptysis. AB - OBJECTIVE: The aim of this study was to determine the incidence and etiology of pulmonary artery pseudoaneurysms in patients undergoing bronchial angiography for massive hemoptysis and to assess patient outcome after the embolization of these pseudoaneurysms. CONCLUSION: Peripheral pulmonary artery pseudoaneurysms occur in up to 11% of patients undergoing bronchial angiography for hemoptysis. These are often most easily appreciated on bronchial and/or nonbronchial systemic arterial angiograms because of complete reversal of flow in pulmonary artery branches in the diseased lung. Embolization of bronchial and nonbronchial systemic arteries alone may not be sufficient therapy to control hemoptysis, and occlusion of the pseudoaneurysm itself via a pulmonary artery approach is recommended. PMID- 15788607 TI - BI-RADS for sonography: positive and negative predictive values of sonographic features. AB - OBJECTIVE: The purpose of this study was to assess the positive predictive value (PPV) and negative predictive value (NPV) of features described in the new sonographic BI-RADS lexicon for evaluating solid masses with known histologic diagnoses. MATERIALS AND METHODS: Sonograms of 403 solid lesions were analyzed by one of three dedicated breast radiologists. Each lesion was described using features from the sonographic BI-RADS lexicon. Lesion description and biopsy results were correlated. PPV and NPV were calculated. RESULTS: Histologic results showed that 141 (35%) of 403 masses were malignant. Sonographic BI-RADS descriptors showing high predictive value for malignancy include spiculated margin (86%, 19/22), irregular shape (62%, 102/164), and nonparallel orientation (69%, 75/109). Sonographic BI-RADS descriptors highly predictive of benign lesions include circumscribed margin (90%, 160/178), parallel orientation (78%, 228/294), and oval shape (84%, 200/237). For the sonographic BI-RADS features of mass margin, shape, orientation, lesion boundary, echo pattern, and posterior acoustic features, descriptors chosen were significantly (p < 0.001) different for malignant and benign masses. CONCLUSION: Descriptors from the new sonographic BI-RADS lexicon can be useful in differentiating benign from malignant solid masses. PMID- 15788608 TI - Comparison of FDG PET and SPECT for detection of bone metastases in breast cancer. AB - OBJECTIVE: The purpose of our study was to evaluate the efficacy of FDG PET and bone SPECT for diagnosing bone metastases in breast cancer. SUBJECTS AND METHODS: The study was a prospective series of 15 patients with breast cancer who underwent both PET and bone scanning with SPECT. Comparison was performed on a lesion-by-lesion analysis. MDCT, MRI, and the patient's clinical course were used as references. RESULTS: In the lesion-by-lesion analysis (n = 900), the sensitivity for diagnosing bone metastases was 85% for SPECT and 17% for PET, specificity was 99% for SPECT and 100% for PET, and accuracy was 96% for SPECT and 85% for PET. In the statistical analysis, bone SPECT was significantly superior to FDG PET for its sensitivity (p < 0.0001) and accuracy (p < 0.0001). No statistically significant difference was seen with regard to specificity. When classifying the bone metastases as osteoblastic or osteolytic, bone scanning classified 92% of metastases as osteoblastic and 35% of metastases as osteolytic, whereas PET classified 6% of metastases as osteoblastic and 90% of metastases as osteolytic. CONCLUSION: Bone SPECT is superior to FDG PET in detecting bone metastases in breast cancer. The sensitivity of osteoblastic lesions is limited with FDG PET. Surveillance of metastatic spread to the skeleton in breast cancer patients based on FDG PET alone is not possible. PMID- 15788609 TI - MRI of metaplastic carcinoma of the breast. AB - OBJECTIVE: The purpose of our study was to describe the MRI findings of metaplastic carcinoma of the breast and to compare those findings with underlying histopathologic features. CONCLUSION: Metaplastic carcinoma of the breast shows high signal intensity on T2-weighted MRI. This finding is related to the necrotic component of the tumor and may be useful for preoperative diagnosis of metaplastic carcinoma of the breast, although it must be differentiated from mucinous carcinoma and, less frequently, necrotic infiltrating ductal carcinoma. PMID- 15788610 TI - Mammography and sonography of pathologically proven adrenal cortical carcinoma metastatic to the breast. PMID- 15788611 TI - How old is this fracture? Radiologic dating of fractures in children: a systematic review. AB - OBJECTIVE: We conducted a systematic review of the literature to define the evidence for radiologic dating of fractures in children in the context of child protection. CONCLUSION: Radiologic dating of fractures is an inexact science. Most radiologists date fractures on the basis of their personal clinical experience, and the literature provides little consistent data to act as a resource. There is an urgent need for research to validate the criteria used in the radiologic dating of fractures in children younger than 5 years. PMID- 15788612 TI - Color Doppler sonography of normal and torsed testicular appendages in children. AB - OBJECTIVE: The purpose of our study was to determine the size and appearance of normal and torsed testicular appendages on color Doppler sonography. SUBJECTS AND METHODS: Thirty-three patients with acute scrotal pain underwent gray-scale and color Doppler sonography and subsequent surgery. Twenty-two patients had torsion of the appendix testis, six had epididymitis, three had torsion of the testis, and two had torsion of the appendix epididymidis. The testicular appendages of 30 patients and the testes of three were excised and sent to the laboratory for histopathologic examination. RESULTS: A testicular appendage was identified on color Doppler sonography in 23 patients (21 patients with torsed appendages and two patients with epididymitis). The torsed appendixes testis measured from 4.1 to 16.3 mm, and the normal appendixes in the two patients with epididymitis measured 4.1 and 5.6 mm. With the visualization of an appendix larger than 5.6 mm as the sonographic diagnostic criterion for torsion of a testicular appendage, sensitivity was 68.2% (95% confidence interval (CI), 45.1-85.3%), and specificity was 100% (CI, 74.7-100%). CONCLUSION: The identification of a testicular appendage larger than 5.6 mm is suggestive of torsion. Therefore, depending on the patients' clinical conditions, these cases can be treated conservatively when an appendage larger than 5.6 mm is identified. PMID- 15788613 TI - PET/CT in the evaluation of childhood sarcomas. AB - OBJECTIVE: Our objective was to review our preliminary experience with PET/CT in evaluating childhood sarcomas including rhabdomyosarcoma (n = 28), the Ewing's sarcoma family of tumors (n = 14), nonrhabdomyosarcoma soft-tissue sarcoma (n = 9), osteosarcoma (n = 8), chondrosarcoma (n = 1), and embryonal sarcoma (n = 1). CONCLUSION: We found PET/CT useful in depicting an unknown primary rhabdomyosarcoma and detecting unsuspected and unusual metastatic sites of childhood sarcomas. It was useful in monitoring response to chemotherapy, radiation therapy, and radiofrequency ablation and aided the postoperative evaluation of tumor resection sites. PMID- 15788614 TI - Pancreatic cystosis in children and young adults with cystic fibrosis: sonographic, CT, and MRI findings. AB - OBJECTIVE: We sought to analyze the sonographic, CT, and MRI findings of pancreatic cystosis, an unusual form of pancreatic involvement in cystic fibrosis (CF) that is characterized by macrocysts of different sizes distributed throughout the gland. CONCLUSION: CT and MRI essentially confirmed the findings of sonography in all cases, providing no relevant additional information about the nature, internal structure, or content of the cysts. When pancreatic macrocysts are clearly shown on sonography in asymptomatic patients with CF, no additional imaging is warranted. PMID- 15788615 TI - Infantile myofibromatosis of the posterior fossa. PMID- 15788616 TI - Arterialization of cerebral veins on dynamic MDCT angiography: a possible sign of a dural arteriovenous fistula. AB - OBJECTIVE: MDCT angiography allows fast imaging of the cerebral vessels, and its potential as a noninvasive technique to detect vascular abnormalities on the basis of morphologic changes is well established. We analyzed vascular enhancement patterns of cerebral venous structures on MDCT angiography, which enabled us to diagnose dural arteriovenous fistula. CONCLUSION: MDCT angiography performed during an early arterial phase showed asymmetrically higher contrast intensity in the transverse or sigmoid sinus, or both, in five patients. In all patients, digital subtraction angiography confirmed the presence of a dural arteriovenous fistula on the side on which the higher contrast intensity appeared. Radiologists should actively look for this sign in the imaging workup of patients presenting with nonspecific symptoms that might be related to a dural arteriovenous fistula. PMID- 15788618 TI - Diffusion-weighted MR microimaging of the lacrimal glands in patients with Sjogren's syndrome. AB - OBJECTIVE: The purpose of this study was to detect quantitative diffusion weighted abnormalities in the lacrimal glands of patients with Sjogren's syndrome. MATERIALS AND METHODS: Diffusion-weighted MRI was performed on 31 healthy volunteers and 11 Sjogren's syndrome patients with impaired lacrimal function. The volunteers and patients underwent MRI with a single-shot spin-echo echo-planar technique using a 47-mm microscopy coil. The apparent diffusion coefficient (ADC) of the lacrimal and parotid glands was obtained with b factors of 500 and 1,000 sec/mm(2). T1-weighted and fat-suppressed T2-weighted MR microscopic images were also obtained to evaluate the gland morphology and signals. RESULTS: MR microscopy provided high-resolution images of the lacrimal glands that enabled ADC measurements. The ADCs of the normal lacrimal glands showed no significant sex- or age-related changes. The ADCs for the lacrimal glands were significantly higher than those of the parotid glands in the same subjects (mean +/- SD, 891 +/- 103 vs 703 +/- 84 x 10(-6) mm(2)/sec, respectively; p < 0.0001, Mann-Whitney U test). We found that ADCs of the lacrimal glands in Sjogren's syndrome patients were significantly lower than those from the normal glands of age-matched healthy volunteers (736 +/- 34 vs 923 +/- 84 x 10(-6) mm(2)/sec; p < 0.0001, Mann-Whitney U test). CONCLUSION: These findings suggest that the measurement of ADCs may be a useful tool to assess abnormalities of the lacrimal glands in patients with Sjogren's syndrome. PMID- 15788617 TI - Imaging of cerebral isolated cortical vein thrombosis. PMID- 15788619 TI - Rational diagnosis of squamous cell carcinoma of the head and neck region: comparative evaluation of CT, MRI, and 18FDG PET. AB - OBJECTIVE: We sought to evaluate the efficiency of (18)FDG PET, CT, and MRI for the preoperative staging of squamous cell carcinoma (SCC) of the head and neck region. CONCLUSION: MRI is recommended as the method of choice in the preoperative evaluation of SCC of the oral cavity and the oropharynx. PET can provide relevant diagnostic information in case of equivocal findings by MRI or CT. Routine use of PET, however, does not appear to be necessary if optimized MRI is available. PMID- 15788620 TI - Management of end hole in placement of port-catheter system for continuous hepatic arterial infusion chemotherapy using the fixed catheter tip method. AB - OBJECTIVE: The purpose of our study was to compare persistent hepatofugal blood flow in the gastroduodenal artery after implanting a port-catheter system for repeated hepatic arterial infusion chemotherapy using either the original or the modified fixed catheter tip method. With the original method the lumen of the catheter tip is closed with a microcoil; with the modified method it is left open. Persistent hepatofugal blood flow can induce reactive gastric or duodenal mucosal lesions. MATERIALS AND METHODS: A port-catheter system with the catheter tip fixed to the gastroduodenal artery by embolic agents was percutaneously implanted in 156 patients (102 men, 54 women; mean age, 63.2 years) with unresectable liver cancer. In 98 patients the original method was used, and in 58 patients the modified method was used. Existence of persistent blood flow beyond the indwelling catheter tip as shown on arteriography via the port performed immediately and 2-10 days after port-catheter placement was compared between these two groups. RESULTS: In all cases, percutaneous port-catheter placement was successfully performed. In one (1.0%) of 98 procedures involving the original method, the gastroduodenal artery was detected on arteriography just after implantation, compared with 23 (39.7%) of 58 procedures using the modified method. However, arteriography performed 2-10 days (mean, 5.02 days) after implantation detected the gastroduodenal artery in only one case. CONCLUSION: This retrospective study indicates that closure of the end hole appears to occur spontaneously shortly after implantation. Thus, such closure is not always necessary to avoid persistent hepatofugal blood flow in the gastroduodenal artery. PMID- 15788622 TI - Radiofrequency tumor ablation: insight into improved efficacy using computer modeling. AB - OBJECTIVE: To use computer modeling of the Bio-Heat equation to demonstrate factors influencing RF ablation tissue heating. CONCLUSION: Computer modeling demonstrates the importance of energy deposition, tumor and background tissue electrical and thermal conductivity, and perfusion on RF ablation outcomes. PMID- 15788621 TI - Balloon-occluded retrograde transvenous obliteration of gastric varices with gastrorenal shunt: long-term follow-up in 78 patients. AB - OBJECTIVE: Our aim was to evaluate the long-term clinical results after balloon occluded retrograde transvenous obliteration (B-RTO) for gastric varices with spontaneous gastrorenal shunt. MATERIALS AND METHODS: A total of 78 patients with cirrhosis and with gastric varices, successfully treated by B-RTO, were enrolled in this study. Recurrence and bleeding of gastric varices and worsening of esophageal varices were endoscopically evaluated. Univariate and multivariate analyses were used to assess the prognostic factors for worsening of esophageal varices and survival. RESULTS: Recurrence of gastric varices was found in two patients; the 5-year recurrence rate was 2.7%. Bleeding of gastric varices occurred in only one patient after B-RTO; the 5-year bleeding rate was 1.5%. Worsening of esophageal varices was observed in 29 patients, and the worsening rates at 1, 3, and 5 years were 27%, 58%, and 66%, respectively. These esophageal varices were endoscopically treated to prevent rupture. Multivariate analysis showed the presence of esophageal varices before B-RTO was a prognostic factor for worsening (relative risk, 4.956). At a median follow-up of 700 days (range, 137-2,339 days), the survival rates at 1, 3, and 5 years were 93%, 76%, and 54%, respectively. The prognostic factors associated with survival were presence of hepatocellular carcinoma (relative risk, 24.342) and the Child-Pugh classification (relative risk, 5.780). CONCLUSION: B-RTO is an effective method for gastric varices with gastrorenal shunt and provides lower recurrence and bleeding rates. We believe that B-RTO can become a standard treatment for gastric varices with gastrorenal shunt, although treatment of worsened esophageal varices may be necessary after B-RTO. PMID- 15788623 TI - A solution for transferring 35-mm slide collections into a digital teaching-file database system. AB - OBJECTIVE: Our objective was to develop a convenient yet comprehensive process for transferring 35-mm radiology slides into a digital teaching-file system capable of delivering online educational content and serving also as a general purpose digital media repository. CONCLUSION: We believe this approach provides a feasible solution for converting radiologists' educational 35-mm slides into well organized, high-quality digital media suitable for both online education and speaker-led presentations. PMID- 15788624 TI - Projecting PowerPoint presentations with a PDA. AB - OBJECTIVE: This article describes the use of a personal digital assistant (PDA) as a projection device for PowerPoint presentations. CONCLUSION: Advances in hardware and software have made PDA devices capable of storing, accessing, retrieving, and sharing data, and their use is becoming increasingly evident in the health care arena. In this article, I describe a use for the PDA that will further increase its value and efficiency in daily academic and teaching activities. PMID- 15788625 TI - Value of IV contrast medium in subtraction CT angiography. PMID- 15788626 TI - CT pulmonary angiography. PMID- 15788627 TI - Opacification of the inferior vena cava and inspiration-associated artifacts. PMID- 15788628 TI - Diffusion-weighted imaging findings in central skull base osteomyelitis with pharyngeal abscess formation. PMID- 15788629 TI - MDCT of pulmonary embolism after percutaneous vertebroplasty. PMID- 15788637 TI - Primary cell cultures as models of prostate cancer development. AB - This review focuses on primary cultures of human prostatic epithelial cells and their applications as models of normal and malignant biological behavior. Current abilities to culture cells from normal tissues, from premalignant dysplastic lesions (prostatic intraepithelial neoplasia), from primary adenocarcinomas, and from metastases are described. Evidence for representation of the interrelated cells of the normal prostatic epithelium--stem cells, basal epithelial cells, secretory epithelial cells, transit amplifying cells and neuroendocrine cells--in primary cultures is presented. Comparisons between normal and cancer-derived primary cultures are made regarding biological activities relevant to carcinogenesis, such as proliferation, apoptosis, differentiation, senescence, adhesion, migration, invasion, steroid hormone metabolism, other metabolic pathways and angiogenesis. Analyses of tumor suppressor activity, differential gene expression and cytogenetics in primary cultures have revealed changes relevant to prostate cancer progression. Preclinical studies with primary cultures have provided information useful for designing new strategies for chemoprevention, chemotherapy, cytotoxin therapy, radiation therapy, gene therapy and imaging. While the behavior of normal primary cultures is often used as a basis for comparison with established, immortal prostate cancer cell lines, the most informative studies are performed with donor-matched pairs of normal and malignant primary cultures, grown under identical conditions. Challenges that remain to be addressed if the full potential of primary cultures as a model system is to be realized include isolation, culture and characterization of stem cells, improved methodology to induce or maintain a fully differentiated, androgen-responsive phenotype, and identification of cell surface antigens or other markers with which to purify pure populations of live cancer or premalignant cells apart from non-malignant epithelial cells prior to culture. PMID- 15788636 TI - Breast cancer vaccines: maximizing cancer treatment by tapping into host immunity. AB - Optimizing standard treatment modalities for breast cancer has improved the outlook for women afflicted with it, but the fact that 40% still ultimately die from the disease highlights the need for new therapies. Remarkable advances in molecular immunology and biotechnology have created a unique opportunity for developing active vaccination strategies that engage the patient's own immune system in the fight against breast cancer. Early clinical trials have established the safety and bioactivity of some breast cancer vaccine approaches, with a hint of clinical response. They have also highlighted the importance of elucidating the pharmacodynamic interactions between established therapies for breast cancer, such as tamoxifen, aromatase inhibitors, chemotherapy, the HER-2/neu-specific monoclonal antibody trastuzumab (Herceptin), and breast cancer vaccines. Preclinical studies have simultaneously defined the importance of developing targeted approaches for circumventing established immune tolerance to breast cancer during the vaccination process. The first strategies targeting the negative influence of CD4(+)CD25(+)T regulatory cells and the CTLA-4 signaling pathway are just entering clinical testing in combination with tumor vaccines. Developing the most potent approach for activating antitumor immunity while maintaining the efficacy of standard approaches to breast cancer management will ensure that active immunotherapy is successfully integrated into the standard of care. PMID- 15788638 TI - rhTSH-aided radioiodine ablation and treatment of differentiated thyroid carcinoma: a comprehensive review. AB - Traditionally, withdrawal of thyroid hormone has been used to attain the increase in serum TSH concentrations that are believed to optimize the trapping and retention of radioiodine for diagnostic procedures, thyroid remnant ablation and treatment of patients with differentiated thyroid cancer (DTC). However, withdrawal frequently causes clinical hypothyroidism, with resultant cognitive impairment, emotional dysfunction, physical discomfort, health risks in patients who are elderly, frail or have concomitant illness, and impaired quality of life and ability to work. Recombinant human TSH (rhTSH) was developed to provide TSH stimulation without withdrawal of thyroid hormone and the associated morbidity. rhTSH has been approved as an adjunct for diagnostic procedures in patients with DTC, but is currently an experimental aid in thyroid remnant ablation and the treatment of thyroid tumours. In the period 1997-2004, nearly 30 medical centres worldwide have reported on almost 400 patients with DTC who were given rhTSH in preparation for radioiodine ablation of thyroid remnants or treatment of local tumours of metastatic disease. We have analysed and summarized the findings reported in this literature. Ablation aided by the standard course of rhTSH, two consecutive daily injections of 0.9 mg, had success rates better than 84% in 90 patients given radioiodine activities in excess of 4000 MBq. However, when 1110 MBq was administered, success rates were 81.2% in 16 patients given the standard course of rhTSH and 4-day withdrawal of thyroid hormone around the time of radioiodine administration in one study, but 54% in 70 patients in another study. rhTSH-aided treatment of persistent or recurrent local or metastatic cancer, or both, with from one to six courses of radioiodine 1000-19055 MBq, achieved 2% complete remission, 36% partial response and 27% disease stabilization rates, for a 65% clinical benefit rate, in 115 primarily elderly, late-stage patients for whom responses were reported. Twelve of these patients died as a result of progressive disease or were discharged from hospital into hospice care. Generally, rhTSH was very well tolerated. However, in a minority of patients with central nervous system, spinal or bone metastases, or bulky thyroid remnant or neck lesions with or without poor pulmonary reserve, administration of rhTSH, like thyroid hormone withdrawal, was found to stimulate expansion of the tumour, with ensuing compression of key anatomical structures and neurological, respiratory or other clinical complications. The rapid onset, response to glucocorticoids and radiological findings of peritumoural oedema or, less commonly, haemorrhage in the published cases, strongly suggest that the tumour expansion was the result of swelling rather than growth. As in the case of thyroid hormone withdrawal, special attention and glucocorticoid premedication are thus warranted when rhTSH is given to patients known or suspected to have the above characteristics. Dosimetric data suggest that whole-body and whole-blood radioiodine clearance may be faster in euthyroid patients after administration of rhTSH. In theory, the faster clearance could allow, or demand, increased radioiodine activities when rhTSH is used, but clinical data to date suggest that this may be unnecessary. The faster clearance also might result in safety or convenience benefits with the use of rhTSH, such as decreased exposure of extrathyroid areas to radiation, and shorter hospital stays. In conclusion, in preliminary results from open-label studies, both rhTSH-aided tumour ablation and treatment have been well tolerated and have shown efficacy in substantial proportions of patients. rhTSH-aided ablation merits further study. rhTSH-aided treatment may be preferred in patients who are at greater risk of hypothyroid complications from withdrawal of thyroid hormone or are unable to produce sufficient endogenous TSH, and warrants additional investigation in younger patients at earlier stages of thyroid cancer. PMID- 15788639 TI - Identification of RAI3 as a therapeutic target for breast cancer. AB - We have been investigating gene-expression profiles in estrogen receptor (ER) negative breast cancers to identify molecules involved in breast carcinogenesis and to select genes or gene products that might be useful as diagnostic markers or targets for new molecular therapies. Here we report evidence that the gene encoding retinoic acid-induced protein 3 (RAI3) is a potential molecular target for treatment of breast cancers. Using quantitative reverse transcription-PCR (RT PCR), we documented increased expression of RAI3 in 19 of 25 primary breast cancers and in 6 of 11 breast-cancer cell lines examined, by comparison with normal mammary-gland tissue. Treatment of human embryonic kidney (HEK293) cells with siRNA against RAI3 suppressed expression of RAI3 and also suppressed cell growth. Transfection of siRNA into breast-cancer cell lines MCF7 and T47D also suppressed RAI3 mRNA and growth of the cancer cells. Because our data imply that up-regulation of RAI3 function is a frequent feature of breast carcinogenesis, we suggest that selective suppression of signal from RAI3 might hold promise for development of a new strategy for treating breast cancers. PMID- 15788640 TI - Molecular changes associated with the agonist activity of hydroxy-tamoxifen and the hyper-response to estradiol in hydroxy-tamoxifen-resistant breast cancer cell lines. AB - The aim of this study was to explore the pharmacological response to 4-hydroxy tamoxifen (OH-Tam) and to estradiol (E2) in three cell lines: MVLN, a human breast carcinoma cell line derived from MCF-7, and two MVLN-derived OH-Tam resistant (OTR) cell lines, called CL6.8 and CL6.32. The OH-Tam response in the OTR cells was associated with the development of both an agonist activity of the drug on cell proliferation and the resistance of the cells to OH-Tam-induced apoptosis. The OTR cells also developed an increased sensitivity to the E2 growth stimulating activity. To delineate the genes that determine such responses, we combined a mini-array-based gene-selection approach and an extensive real-time quantitative PCR exploration in the MVLN and OTR cell lines exposed to three pharmacological conditions: a 4-day treatment with E2, OH-Tam or both E2 and OH Tam. Compiled data revealed a hyper-response to E2 and a modification of the OH Tam pharmacological response (loss of antagonist action and agonist activity) at the gene-expression level. The proteins encoded by the genes selected in this study have been reported to be involved in the regulation of cell proliferation, cell transformation, DNA repair and apoptosis, or belong to the ErbB/epidermal growth factor receptor-driven pathway. Our data also provide evidence of changes in transcriptional co-regulator expression, elevated mitogen-activated protein kinase activity and increase in the phosphorylation status of estrogen receptor alpha on serine residue 118 in the OTR cell lines, suggesting the possible involvement of such mechanisms in the agonist activity of OH-Tam and/or the hyper response of cells to E2. Taken together, our study should enhance our knowledge of the multifactorial events associated with the development of Tam resistance in two independent cell lines issued from the same selection process and should help in the identification of potential molecular targets for diagnosis or therapy. PMID- 15788641 TI - Effect of small-sized liposomal Adriamycin administered by various routes on a metastatic breast cancer model. AB - The antitumor effects of small-sized liposomal Adriamycin (LADR) administered by various routes were investigated in rabbits bearing well-developed VX2 tumors in the mammary gland. Rabbits received s.c. or i.v., or s.c. combined with i.v., injections of LADR 6 weeks after tumor implantation. The i.v. route showed a significant inhibitory effect on breast tumors and distant metastases. In comparison, metastases in axillary and mediastinal lymph nodes were more efficiently inhibited after s.c. injection. LADR administered concurrently by both the i.v. and s.c. routes produced satisfactory therapeutic activities on both primary breast tumors and metastases in local-regional lymph nodes, lungs and liver, as shown by slowed growth rates, decreased mRNA expression of proliferating cell nuclear antigen, and extensive necrosis and apoptosis of tumor cells. It is concluded that small-sized LADR administered s.c. provides reliable efficacy on lymphatic metastases of breast cancer and that the addition of treatment by the s.c. route to that by the conventional i.v. route can be recommended as a promising procedure to enhance chemotherapeutic effects in patients with metastatic breast cancer. PMID- 15788642 TI - In situ androgen producing enzymes in human prostate cancer. AB - Androgens have been proposed to be actively produced in situ in human prostate cancer. These locally produced androgens have also been considered to play important roles in the pathogenesis and development of prostate cancer. Therefore, it is important to examine the status of this in situ androgen metabolism and/or synthesis in detail in order to improve the clinical response to hormonal therapy in patients diagnosed with prostate cancer. Several studies have previously demonstrated the expression of androgen-producing enzymes such as 5alpha-reductase types 1 and 2, and 17beta-hydroxysteroid dehydrogenase type 5 (17beta-HSD5), in human prostate carcinoma cells. However, their biological significance has remained largely unknown. In this study, we evaluated the immunoreactivities of these steroidogenic enzymes in human prostate cancer obtained from surgery (n = 70), and correlated the findings with clinicopathological features of the patients. 17Beta-HSD5 immunoreactivity was detected in 54 cases (77%), 5alpha-reductase type 1 in 51 cases (73%) and 5alpha reductase type 2 in 39 cases (56%). 5Alpha-reductase type 2 immunoreactivity was significantly correlated with that of androgen receptor (AR), and 17beta-HSD5 positive cases were significantly associated with clinical stage (TNM stage pT3 vs pT2). These data all suggest that androgen-producing enzymes, such as 5alpha reductase type 1 and type 2, and 17beta-HSD5 are expressed in a majority of prostate cancers, and are involved in the local production and actions of androgens in prostate cancers. PMID- 15788643 TI - Independent prognostic role of circulating chromogranin A in prostate cancer patients with hormone-refractory disease. AB - The presence of neuroendocrine (NE) differentiation in the context of predominantly exocrine prostate cancer may play a key role in androgen independent tumor growth. The prognostic significance of plasma chromogranin A (CgA) was assessed in a series of consecutive prostate cancer patients with hormone-refractory disease. One hundred and eight patients with newly diagnosed hormone-refractory prostate cancer entered the study. Plasma CgA levels and other biochemical parameters, such as serum prostate specific antigen, serum alkaline phosphatase, serum lactate dehydrogenase, serum albumin and hemoglobin concentration, were measured at baseline (i.e. when hormone refractoriness occurred) and their prognostic role was evaluated together with patient performance status, Gleason score (at diagnosis of prostate cancer) and the presence of visceral metastases. Furthermore, plasma CgA was prospectively evaluated in 50 patients undergoing chemotherapy. At baseline, 45 patients (43.3%) showed elevated CgA values. Plasma CgA negatively correlated with survival, either in univariate analysis (P=0.008) or in multivariate analysis, after adjusting for previously mentioned prognostic parameters (P<0.05). In the patient subset undergoing chemotherapy, median CgA (range) values were 13.3 (3.0 141.0) U/l at baseline, 19.1 (3.0-486.0) U/l after 3 months, 20.8 (3.0-702.0) U/l after 6 months and 39.4 (3.0-414.0) U/l after 9 months (P<0.01). The corresponding supranormal rates were 17/50 (34%), 23/50 (46%), 26/50 (52%) and 34/50 (68%) respectively (P<0.005). Elevated plasma CgA levels are frequently observed in prostate cancer patients with hormone-refractory disease and correlate with poor prognosis. NE differentiation in hormone-refractory patients is a time-dependent phenomenon and is not influenced by conventional antineoplastic treatments. PMID- 15788645 TI - Expression profiling of androgen-dependent and -independent LNCaP cells: EGF versus androgen signalling. AB - Prostate cancer development often includes a shift from androgen-dependent to androgen-independent growth. It is hypothesized that, during this transition, growth factors like the epidermal growth factor (EGF) gain importance as activators of tumour cell proliferation. To study this, androgen- and EGF regulation of growth and gene-expression was analysed in the androgen-dependent human prostate cancer cell line LNCaP-FGC (FGC) and its androgen-independent derivative line LNCaP-LNO (LNO). It was observed that androgen-dependent FGC cells require exposure to either androgens or EGF to proliferate. This is in contrast to androgen-independent LNO cells that showed significant proliferation in medium depleted of androgens and growth factors. Gene expression data were obtained for the androgen-dependent FGC and androgen-independent LNO cells cultured in the presence or absence of androgens (synthetic R1881) or EGF for different time periods. Expression profiling showed that many cell cycle genes, including a number of androgen- and EGF-regulated genes, are constitutively activated in androgen-independent LNO cells. Furthermore, the overlap between changes in gene expression activated by androgen and EGF receptor signalling pathways was found to be very high (75%). These results partly explain why androgen-independent LNO cells can proliferate in the absence of androgenic stimulation. However, possibly other, so far unknown, signal transduction pathways that induce and maintain proliferation, have also been activated. PMID- 15788644 TI - Signal transduction pathways in androgen-dependent and -independent prostate cancer cell proliferation. AB - In a previous report, we showed that increased activation of Akt, a downstream effector of phosphoinositide 3-kinase (PI3K) together with decreased activation of extracellular-signal-regulated kinase (ERK), a member of the mitogen-activated protein kinase (MAPK) family, predicted poor clinical outcome in prostate cancer (Kreisberg et al. 2004 Cancer Research 64 5232-5236). We now show that Akt activation, but not ERK activation, is correlated with proliferation in human prostate tumors as estimated by the expression of the cell proliferation antigen Ki67. We verified these results in vitro, using the androgen-dependent prostate cancer cell line LNCaP and its androgen-independent clone C4-2 as models of prostate cancer of good and poor clinical outcome, respectively. C4-2 cells expressed higher Akt activation, lower ERK activation and increased proliferation compared with LNCaP cells, similar to cases of poor clinical outcome. The PI3K inhibitor LY294002, but not the MAPK/ERK kinase inhibitor PD98059, induced growth arrest in both cell lines. Transient transfection with constitutively active Akt increased proliferation while dominant negative Akt decreased it, thus showing that Akt plays an important role in prostate cancer proliferation. Akt regulates the expression and activation of the androgen receptor. Androgen receptor inhibition with Casodex induced growth arrest in LNCaP cells, but not in C4-2 cells. Another PI3K downstream effector, p70 S6 kinase, requires prior phosphorylation by mammalian target of rapamycin (mTOR) for complete activation. Activation of p70 S6 kinase was higher in C4-2 compared with LNCaP cells. Rapamycin, an mTOR inhibitor, had a growth-inhibitory effect in C4-2 cells, but not in LNCaP cells. Our data suggest a shift from a Casodex-sensitive proliferation pathway in LNCaP cells to a rapamycin-sensitive pathway in C4-2 cells. PMID- 15788646 TI - Aldosterone-producing adrenocortical carcinoma: an unusual cause of Conn's syndrome with an ominous clinical course. AB - Aldosterone-producing adrenocortical carcinoma (APAC) is a rare cause of hypertension often diagnosed late because of paucity of information. Thus, we delineated its clinical course and survival rates based on two cases referred to us that featured diverging clinical courses, and on a scrutiny of the literature since 1955 when the first case of APAC was identified. Data on demography, imaging results, hormonal assessment, histology, and clinical course were extracted independently by the investigators. We included in our database 58 cases, most presenting with Conn's syndrome. Plasma aldosterone levels were on average increased 14-fold; plasma renin activity was suppressed in 55% of cases. The tumor showed extremely variable size and weight, and no gender or side preference. Metastases were present in 10% of all cases at initial diagnosis and in an additional 48% of cases at follow-up. Median survival was 546 days (95% confidence interval (CI): 240-851); median time to either recurrence or death was 212 days (95% CI: 29-395). No clinical or histological signs predicted survival with Cox regression analysis. We concluded that, although an ominous course with a poor survival rate is common, no sign accurately predicts the course of APAC. Thus, molecular studies to identify diagnostic markers of survival are mandatory. PMID- 15788647 TI - Epigenetic analysis of HIC1, CASP8, FLIP, TSP1, DCR1, DCR2, DR4, DR5, KvDMR1, H19 and preferential 11p15.5 maternal-allele loss in von Hippel-Lindau and sporadic phaeochromocytomas. AB - Phaeochromocytoma is a neural-crest-derived tumour that may be a feature of several familial cancer syndromes including von Hippel-Lindau (VHL) disease, multiple endocrine neoplasia type 2 (MEN 2), neurofibromatosis type 1 (NF1) and germline succinate dehydrogenase subunit (SDHB and SDHD) mutations. However the somatic genetic and epigenetic events that occur in phaeochromocytoma tumourigenesis are not well defined. Epigenetic events including de novo promoter methylation of tumour-suppressor genes are frequent in many human neoplasms. As neuroblastoma and phaeochromocytoma are both neural-crest-derived tumours, we postulated that some epigenetic events might be implicated in both tumour types and wished to establish how somatic epigenetic alterations compared in VHL associated and sporadic phaeochromocytomas. We identified frequent aberrant methylation of HIC1 (82%) and CASP8 (31%) in phaeochromocytoma, but both genes were significantly more methylated in VHL phaeochromocytomas than in sporadic cases. Of four tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors analysed, DR4 was most commonly methylated (41%; compared with DcR2 (26%), DcR1 (23%) and DR5 (10%)). Gene methylation patterns in phaeochromocytoma and neuroblastoma did not differ significantly suggesting overlapping mechanisms of tumourigenesis. We also investigated the role of 11p15.5-imprinted genes in phaeochromocytoma. We found that in 10 sporadic and VHL phaeochromocytomas with 11p15.5 allele loss, the patterns of methylation of 11p15.5-differentially methylated regions were consistent with maternal, rather than, paternal chromosome loss in all cases (P<0.001). This suggests that 11p15.5-imprinted genes may be implicated in the pathogenesis of both familial (germline VHL and SDHD mutations) and sporadic phaeochromocytomas. PMID- 15788648 TI - ret/PTC1 and ret/PTC3 in thyroid tumors from Chernobyl liquidators: comparison with sporadic tumors from Ukrainian and French patients. AB - Like children exposed to Chernobyl fallout, the workers who cleaned up after the accident, also known as liquidators, have exhibited an increased incidence of thyroid cancer. A high prevalence of ret/PTC3 rearrangement has been found in pediatric post-Chernobyl thyroid tumors, but this feature has not been investigated in liquidator thyroid tumors. In this study we analyzed the prevalence of ret/PTC1 and ret/PTC3 in thyroid tumors from 21 liquidators, 31 nonirradiated adult Ukrainian patients, and 34 nonirradiated adult French patients. ret rearrangements in carcinomas were found in 83.3% of liquidators, 64.7% of Ukrainian patients, and 42.9% of French patients. The prevalence of ret/PTC1 was statistically similar in the three groups. The prevalence of ret/PTC3 was significantly higher in liquidators than in French patients (P = 0.03) but it was also high in nonirradiated Ukrainian patients who exhibited values intermediate between liquidators and French patients. In adenomas the prevalence of rearrangement was significantly higher in all Ukrainians than in French patients (P = 0.004). Like children exposed to Chernobyl fallout, liquidators showed a high prevalence of ret/PTC3. This finding suggests that irradiation had the same effect regardless of age. However, given the high rate of ret/PTC3 in nonirradiated adult Ukrainians, the possibility of genetic susceptibility or low-level exposure to radiation in that group cannot be excluded. PMID- 15788649 TI - Exploited students? PMID- 15788650 TI - The nucleolus. PMID- 15788651 TI - Fran Balkwill. PMID- 15788652 TI - Procollagen trafficking, processing and fibrillogenesis. AB - Collagen fibrils in the extracellular matrix allow connective tissues such as tendon, skin and bone to withstand tensile forces. The fibrils are indeterminate in length, insoluble and form elaborate three-dimensional arrays that extend over numerous cell lengths. Studies of the molecular basis of collagen fibrillogenesis have provided insight into the trafficking of procollagen (the precursor of collagen) through the cellular secretory pathway, the conversion of procollagen to collagen by the procollagen metalloproteinases, and the directional deposition of fibrils involving the plasma membrane and late secretory pathway. Fibril associated molecules are targeted to the surface of collagen fibrils, and these molecules play an important role in regulating the diameter and interactions between the fibrils. PMID- 15788653 TI - Nuclear import of the histone acetyltransferase complex SAS-I in Saccharomyces cerevisiae. AB - The protein complex SAS-I links histone acetylation to the assembly of repressed chromatin in Saccharomyces cerevisiae. Sas2p, the histone acetyltransferase subunit of SAS-I, forms a complex with Sas4p and Sas5p, which are both required for maximal complex activity. In this study, we found that Sas4p was the central subunit of the SAS-I complex, bridging Sas2p and Sas5p. We demonstrated that the nuclear import of Sas2p and Sas5p was mediated by two karyopherins/importins, Kap123p and Pse1p, and both were associated in vivo with these importins. By contrast, Sas4p was not a substrate of Kap123p or Pse1p, suggesting that the nuclear import of the SAS-I subunits occurred independently of each other. Several other non-essential karyopherins were not involved in the nuclear import of SAS-I subunits. When the putative nuclear localization signal (NLS) of Sas2p was deleted, nuclear accumulation of Sas2p was significantly decreased. By contrast, deletion of the proposed NLS of Sas4p had no influence on its nuclear localization. An unknown signal region was located in the N-terminal domain of Sas5p and was responsible for the nuclear import by Kap123p and Pse1p. We found a striking similarity between the NLS sequences of Sas2p and those of histones H3 and H4, which were recently reported to be further import substrates of Kap123p and Pse1p. A database search based on the aligned consensus sequence revealed potential new import substrates of the Kap123p and Pse1p nuclear import pathways, which are connected to chromatin function. PMID- 15788654 TI - Caspase regulation in non-small cell lung cancer and its potential for therapeutic exploitation. AB - Metastatic non-small cell lung cancer (NSCLC, stages IIIB/IV) is one of the most common and rapidly lethal causes of cancer related mortality worldwide. Efficacy of chemotherapy, the mainstay of treatment, is limited due to resistance in the vast majority of patients. NSCLC cells exhibit intrinsic apoptosis resistance. Understanding the molecular basis of this phenotype is critical, if therapy is to move beyond the therapeutic plateau that has been reached with conventional chemotherapy. Caspases occupy a pivotal position in the final common pathway of apoptosis. Increasing evidence suggests that these proteases are constitutively inhibited in NSCLC. This review discusses current knowledge relating to caspase regulation in NSCLC and highlights novel strategies for reversing the apoptosis resistant phenotype, with potential to accelerate development of effective therapy. PMID- 15788655 TI - Mutations in the tyrosine kinase domain of the epidermal growth factor receptor in non-small cell lung cancer. AB - We evaluated somatic genetic alterations in the kinase domain of the EGFR gene in the tumors of 219 non-small cell lung cancer patients of primarily Caucasian and African American origins. We identified 26 patients (12%) whose tumors had a mutation in the EGFR gene, and 11 (5%) patients carried novel genomic variations consistent with germ-line polymorphisms. All but one mutation were identified in Caucasian patients affected with adenocarcinoma. EGFR mutations were more frequent in women and in nonsmokers, but a significant portion of the affected patients were men (12 of 26) and current or past smokers accounted for half of the patients affected (13 of 26). Screening subjects with EGFR mutations may identify patients whose tumors could respond to targeted therapy using tyrosine kinase inhibitors. PMID- 15788656 TI - Associations and interactions between Ets-1 and Ets-2 and coregulatory proteins, SRC-1, AIB1, and NCoR in breast cancer. AB - PURPOSE: Associations between p160 coactivator proteins and the development of resistance to endocrine treatment have been described. We hypothesized that nuclear receptor coregulatory proteins may interact with nonsteroid receptors. We investigated the mitogen-activated protein kinase-activated transcription factors, Ets, as possible interaction proteins for the coactivators SRC-1 and AIB1 and the corepressor NCoR in human breast cancer. EXPERIMENTAL DESIGN: Expression and coexpression of Ets and the coregulatory proteins was investigated using immunohistochemistry and immunofluorescence in a cohort of breast tumor patients (N = 134). Protein expression, protein-DNA interactions and protein protein interactions were assessed using Western blot, electromobility shift, and coimmunoprecipitation analysis, respectively. RESULTS: Ets-1 and Ets-2 associated with reduced disease-free survival (P < 0.0292, P < 0.0001, respectively), whereas NCoR was a positive prognostic indicator (P < 0.0297). Up-regulation of Ets-1 protein expression in cell cultures derived from patient tumors in the presence of growth factors associated with tumor grade (P < 0.0013; n = 28). In primary breast tumor cell cultures and in the SKBR3 breast cell line, growth factors induced interaction between Ets and their DNA response element, induced recruitment of coactivators to the transcription factor-DNA complex, and up regulated protein expression of HER2. Ets-1 and Ets-2 interacted with the coregulators under basal conditions, and growth factors up-regulated Ets-2 interaction with SRC-1 and AIB1. Coexpression of Ets-2 and SRC-1 significantly associated with the rate of recurrence and HER expression, compared with patients who expressed Ets-2 but not SRC-1 (P < 0.0001 and P < 0.0001, respectively). CONCLUSIONS: These data describe associations and interactions between nonsteroid transcription factors and coregulatory proteins in human breast cancer. PMID- 15788657 TI - Histologic type, organ of origin, and Wnt pathway status: effect on gene expression in ovarian and uterine carcinomas. AB - PURPOSE: Ovarian and uterine carcinomas manifest several differentiation patterns resembling those seen in nonneoplastic epithelia of the gynecologic tract. Specific oncogene and tumor suppressor gene defects have been associated with particular differentiation patterns in carcinomas arising in either the uterus or ovary. For instance, ovarian and uterine carcinomas with endometrioid differentiation frequently show beta-catenin mutations. Whereas type of differentiation is considered in the treatment of uterine carcinomas, it does not presently contribute to decisions about treatment of ovarian carcinomas. A widely accepted view is that the accumulation of specific gene defects and gene expression changes underlies phenotypic traits of cancers, including their response to treatment. EXPERIMENTAL DESIGN: Using oligonucleotide microarrays to assess gene expression in 103 primary ovarian and uterine carcinomas, we sought to address whether organ of origin or type of differentiation (histotype; endometrioid versus serous) had a more substantial effect on gene expression patterns. RESULTS: We found that effects on gene expression due to organ of origin and histotype are similar in magnitude and are parallel in that organ effects are similar in the two histotypes and histotype effects are similar in the two organs. In addition, ovarian and uterine endometrioid adenocarcinomas with beta-catenin defects show a common gene expression signature largely distinct from that seen in tumors lacking such defects. CONCLUSIONS: Our results illustrate how organ of origin, type of differentiation, and specific molecular defects all contribute to gene expression in the most common types of ovarian and uterine cancers. The findings also imply gene expression data will be of value for stratifying ovarian cancer patients for new treatment approaches. PMID- 15788658 TI - Transfer of the sFLT-1 gene in Morris hepatoma results in decreased growth and perfusion and induction of genes associated with stress response. AB - PURPOSE: Inhibition of tumor angiogenesis is emerging as a promising target in the treatment of malignancies. Therefore, monitoring of antiangiogenic approaches with functional imaging and histomorphometrical analyses are desirable to evaluate the biological effects caused by this treatment modality. EXPERIMENTAL DESIGN: Using a bicistronic retroviral vector for transfer of the soluble receptor for the vascular endothelial growth factor (sFLT) hepatoma (MH3924A) cell lines with sFLT expression were generated. In human umbilical vein endothelial cells cultured with conditioned medium of sFLT-expressing hepatoma cells, the inhibitory action of secreted sFLT was determined using a Coulter counter and a thymidine incorporation assay. Furthermore, in vivo experiments were done to measure the effects on tumor growth and perfusion. Finally, the tumors were examined by immunohistochemistry (including computer-assisted morphometry) and DNA chip analysis. RESULTS: Stable sFLT-expressing hepatoma cells inhibited endothelial cell proliferation in vitro. In vivo, growth and perfusion, as measured by H(2)(15)O positron emission tomography, were reduced in genetically modified tumors. However, the immunohistochemically quantified microvascularization and macrovascularization, as indicated by CD31- and alpha actin-positive area, revealed no significant changes, whereas the number of apoptotic cells was increased in sFLT-expressing tumors, although not significantly. DNA chip analysis of tumors with gene transfer showed an increase of genes related to apoptosis, signal transduction, and oxidative stress. CONCLUSION: Our results suggest that sFLT expression inhibits tumor growth and perfusion and enhances expression of apoptosis-related genes in this model. Enhanced expression of genes for signal transduction, stress, and metabolism indicates tumor defense reactions. PMID- 15788659 TI - Morphology of tumor cell nuclei is significantly related with survival time of patients with glioblastomas. AB - PURPOSE: To investigate whether histomorphology of tumor cell nuclei has a significant and independent relation to survival time of patients with glioblastomas. EXPERIMENTAL DESIGN: Seventy-two tumors from 72 patients were investigated by means of digital image analysis. Proliferating and nonproliferating nuclei were separately measured and parameters of nuclear size, shape, texture, and spatial relationships (topometric parameters) were detected. Survival analysis was done regarding morphometric data together with the patients' age, the amount of resection (total or subtotal), and the classification of the tumor as a "primary" (de novo) or "secondary" glioblastoma. RESULTS: The overall relation of all morphometric data to the time of survival was highly significant (Cox analysis, P < 0.0001). Apart from the extent of surgical resection, parameters of nuclear shape and topometric variables, such as the distance between two nuclei lying nearest to each other, showed an independent and significant relation to survival time. The patients' age had also a significant but comparably slight relation to survival time. CONCLUSIONS: The morphology of tumor cell nuclei, as represented by morphometric data, shows a significant relation to survival time of patients with glioblastomas. This relation is statistically independent from the amount of surgical resection, from the patients' age and from the classification of the glioblastoma as being primary or secondary. The results support the view that histomorphometry of tumor cell nuclei is a valuable prognostic marker for patients with glioblastomas. We believe that such a marker ought to be incorporated into the formation of individual therapeutic decisions. PMID- 15788660 TI - Gene expression profiles predict early relapse in ovarian cancer after platinum paclitaxel chemotherapy. AB - PURPOSE: Women with advanced epithelial ovarian cancer are routinely treated with platinum-paclitaxel chemotherapy following cytoreductive surgery, yet only approximately 20% achieve long-term disease-free survival. We hypothesized that differences in gene expression before treatment could distinguish patients with short versus long time to recurrence after administration of platinum-paclitaxel combination chemotherapy. EXPERIMENTAL DESIGN: To test this hypothesis, gene expression profiling of 79 primary surgically resected tumors from women with advanced-stage, high-grade epithelial ovarian cancer was done using cDNA microarrays containing 30,721 genes. Supervised learning algorithms were applied in an effort to develop a binary classifier that could discriminate women at risk for early (< or =21 months) versus late (>21 months) relapse after initial chemotherapy. RESULTS: A 14-gene predictive model was developed using a set of training samples (n = 51) and subsequently tested using an independent set of test samples (n = 28). This model correctly predicted the outcome of 24 of the 28 test samples (86% accuracy) with 95% positive predictive value for early relapse. CONCLUSIONS: Predictive markers for early recurrence can be identified for platinum-paclitaxel combination chemotherapy in primary ovarian carcinoma. The proposed 14-gene model requires further validation. PMID- 15788661 TI - Distinct hypermethylation profile of primary breast cancer is associated with sentinel lymph node metastasis. AB - PURPOSE: Gene promoter region hypermethylation is a significant event in primary breast cancer. However, its impact on tumor progression and potential predictive implications remain relatively unknown. EXPERIMENTAL DESIGN: We conducted hypermethylation profiling of 151 primary breast tumors with association to known prognostic factors in breast cancer using methylation-specific PCR for six known tumor suppressor and related genes: RASSF1A, APC, TWIST, CDH1, GSTP1, and RAR beta2. Furthermore, correlation with sentinel lymph node (SLN) tumor status was assessed as it represents the earliest stage of metastasis that is readily detected. Hypermethylation for any one gene was identified in 147 (97%) of 151 primary breast tumors. The most frequently hypermethylated gene was RASSF1A (81%). RESULTS: Hypermethylation of the CDH1 was significantly associated with primary breast tumors demonstrating lymphovascular invasion (P = 0.008), infiltrating ductal histology (P = 0.03), and negative for the estrogen receptor (P = 0.005), whereas RASSF1A and RAR-beta2 gene hypermethylation were significantly more common in estrogen receptor-positive (P < 0.001) and human epidermal growth factor receptor 2-positive (P < 0.001) tumors, respectively. In multivariate analysis, hypermethylation of GSTP1 and/or RAR-beta2 was significantly associated with patients having macroscopic SLN metastasis compared with those with microscopic or no sentinel node metastasis (odds ratio, 4.59; 95% confidence interval, 2.02-10.4; P < 0.001). In paired SLN metastasis, CDH1 was the most frequently methylated gene (90%) and provides evidence in patients corroborating its role in the clinical development of metastasis. CONCLUSION: Hypermethylation profiling of primary breast tumors is significantly associated with known pathologic prognostic factors and may have additional clinical and pathologic utility for assessing patient prognosis and predicting early regional metastasis. PMID- 15788662 TI - Absence of HER4 expression predicts recurrence of ductal carcinoma in situ of the breast. AB - The type 1 tyrosine kinase receptor HER2 (c-erbB2/neu) is associated with resistance to hormone therapy and poor survival in invasive breast cancer, whereas HER4 expression is associated with endocrine responsiveness. Patterns of tyrosine kinase receptor coexpression may aid prediction of recurrence risk after surgery for ductal carcinoma in situ (DCIS). Women who had undergone surgery for pure DCIS were studied. Out of 129 primary tumors, 39 had recurred and 90 had not recurred after 5 years of follow-up. Primary tumors were compared for HER2, HER3, and HER4, estrogen receptor, and Ki67 by immunohistochemistry. HER2 was expressed in 58%, HER3 in 49%, and HER4 in 63% of nonrecurrent DCIS, compared with HER2 expression in 82% (P = 0.008), HER3 expression in 71% (P = 0.04), and HER4 expression in 36% (P = 0.004) in DCIS that subsequently recurred. Dually expressing HER2/4 DCIS was more likely to be estrogen receptor positive than HER2 only-expressing DCIS (73% versus 53%; P = 0.05). HER2 expression was associated with a higher percentage and HER4 expression a significantly lower percentage of proliferating DCIS cells (median, 13.8% versus 8.4%; P = 0.001). Coexpression of HER2 with HER4 was associated with reduced recurrence compared with HER2-only positive DCIS (P = 0.003). This association remained significant when analyzing only high nuclear-grade DCIS (P = 0.015). Low nuclear grade, low proliferation rate and presence of HER4 expression were independent predictors of nonrecurrence. Potentially, HER4 expression may identify women who could avoid radiotherapy after breast-conserving surgery for DCIS. PMID- 15788663 TI - Association of NCOA3 polymorphisms with breast cancer risk. AB - The nuclear receptor coactivator 3 (NCOA3, also known as AIB1) is a coactivator of nuclear receptors like the estrogen receptor. NCOA3 is overexpressed in approximately 60% of primary human breast tumors, and high levels of NCOA3 expression are associated with tamoxifen resistance and worse survival rate. In contrast, NCOA3 deficiency suppresses v-Ha-ras-induced breast cancer initiation and progression in mice. Here, we analyzed the influence of NCOA3 coding single nucleotide polymorphisms on breast cancer risk by performing a case-control study using a German and a Polish study population and identified an association between NCOA3 polymorphisms and breast cancer. A joint analysis of the German and the Polish study population revealed a significant protective effect for the 1758G>C (Q586H) and 2880A>G (T960T) variants. In addition, haplotype analysis showed a protective effect of the 1758C-2880A and 1758G-2880G haplotypes (odds ratio 0.79; 95% confidence interval, 0.67-0.93; P = 0.004). Because of the impact of NCOA3 in antiestrogen therapy resistance, these polymorphisms might also influence therapy outcome in breast cancer. PMID- 15788664 TI - Glutathione S-transferase P1 genotype and prognosis in Hodgkin's lymphoma. AB - PURPOSE: Glutathione S-transferase P1 (GSTP1) is a member of the GST enzyme superfamily that is important for the detoxification of several cytotoxic drugs and their by-products. A single nucleotide polymorphism results in the substitution of isoleucine (Ile) to valine (Val) at codon 105, causing a metabolically less active variant of the enzyme. We assessed the impact of the GSTP1 codon 105 genotype on treatment outcome in patients with Hodgkin's lymphoma. EXPERIMENTAL DESIGN: The Ile(105)Val polymorphism in the GSTP1 gene was analyzed using a PCR-RFLP technique. Ninety-seven patients with Hodgkin's lymphoma were included and associations with patient characteristics and treatment outcome were analyzed. RESULTS: The GSTP1 Ile(105)Val polymorphism was associated in a dose-dependent fashion with an improved failure-free survival in patients with Hodgkin's lymphoma (P = 0.02). The probability of 5-year survival for patients homozygous for the (105)Val/(105)Val GSTP1 genotype was 100%, for heterozygous patients 74% (95% confidence interval, 56-85), and for patients homozygous for the (105)Ile/(105)Ile genotype 43% (95% confidence interval, 23 61). The Cox multivariate analysis showed that GSTP1 codon 105 genotype was an independent prognostic factor. CONCLUSIONS: The GSTP1 genotype predicts clinical outcome in patients with Hodgkin's lymphoma. PMID- 15788665 TI - Microsatellite instability testing in colorectal carcinoma: choice of markers affects sensitivity of detection of mismatch repair-deficient tumors. AB - PURPOSE: Microsatellite instability (MSI) is found in 10% to 15% of sporadic colorectal tumors and is usually caused by defects in DNA mismatch repair (MMR). In 1997, a panel of microsatellite markers including mononucleotide and dinucleotide repeats was recommended by a National Cancer Institute workshop on MSI. We investigated the relationship between instability of these markers and MMR protein expression in a cohort of sporadic colorectal cancer patients. EXPERIMENTAL DESIGN: Paraffin sections of normal and tumor tissue from 262 colorectal cancer patients were examined for MSI status by PCR amplification and for MMR protein expression using antibodies against hMLH1, hPMS2, hMSH2, and hMSH6. RESULTS: Twenty-six (10%) of the patients studied had tumors with a high level of MSI (MSI-H). The frequencies of MSI were the same in African-American and Caucasian patients. Each of the MSI-H tumors had mutations in both mononucleotide and dinucleotide repeats and had loss of MMR protein expression, as did two tumors that had low levels of MSI (MSI-L). These two MSI-L tumors exhibited mutations in mononucleotide repeats only, whereas eight of the other nine MSI-L tumors had mutations in just a single dinucleotide repeat. There was not a statistically significant difference in outcomes between patients whose tumors were MMR-positive or MMR-negative, although there was a slight trend toward improved survival among those with MMR-deficient tumors. CONCLUSIONS: The choice of microsatellite markers is important for MSI testing. Examination of mononucleotide repeats is sufficient for detection of tumors with MMR defects, whereas instability only in dinucleotides is characteristic of MSI-L/MMR-positive tumors. PMID- 15788666 TI - Aromatase expression in stromal cells of endometrioid endometrial cancer correlates with poor survival. AB - PURPOSE AND EXPERIMENTAL DESIGN: To assess the prognostic significance of intratumoral aromatase in endometrioid endometrial cancer, sections from 55 patients with endometrial cancer were evaluated for expression of aromatase using immunohistochemistry, and the correlation between aromatase expression and clinicopathologic parameters were analyzed. RESULTS: Immunohistochemical staining for aromatase was positive for 32 (58%), 20 (36%), and 19 (34%) patients in cancer epithelial cells, stromal cells, and myometrial cells around the flank invasion, respectively. In situ hybridization also detected aromatase mRNA in all three types of cells. RT-PCR analysis revealed that aromatase mRNA was 2.5 +/- 1.0 amol/mug total RNA (mean +/- SE; n = 7) in tumor tissue. Western blot analysis detected the expected aromatase protein size of 58 kDa in cancer tissues more abundantly than in cancer-free endometrium (n = 3). The immunoreactivity in stromal cells correlated positively with advanced surgical stage and poor survival. Survival analysis revealed that the immunoreactivity of stromal cells was a significant prognostic factor, independent of histologic grade, muscular invasion, and lymph node metastasis, but dependent on surgical stage. By contrast, the immunoreactivity of aromatase both in cancer epithelial cells and myometrial cells did not correlate with prognosis. CONCLUSIONS: To the best of our knowledge, this is the first evidence associating intratumoral aromatase expression in stromal cells and poor survival in endometrioid endometrial cancer. This positive linkage indicates that local expression of aromatase plays a role in tumor progression through the formation of in situ estrogens. In situ expression of aromatase may offer a potential target for management of endometrial cancers. PMID- 15788667 TI - Generation of PRL-3- and PRL-1-specific monoclonal antibodies as potential diagnostic markers for cancer metastases. AB - PURPOSE: The PRL-3 mRNA is consistently elevated in metastatic samples derived from colorectal cancers. We sought to generate a specific PRL-3 monoclonal antibody (mAb) that might serve as a potential diagnostic marker for colorectal cancer metastasis. EXPERIMENTAL DESIGN: PRL-3 is one of three members (PRL-1, PRL 2, and PRL-3) in a unique protein-tyrosine phosphatase family. Because the three PRLs are 76% to 87% identical in their amino acid sequences, it poses a great challenge to obtain mAbs that are specific for respective phosphatase of regenerating liver (PRL) but not for the other two in the family. We screened over 1,400 hybridoma clones to generate mAbs specific to each PRL member. RESULTS: We obtained two hybridoma clones specifically against PRL-3 and another two clones specifically against PRL-1. These antibodies had been evaluated by several critical tests to show their own specificities and applications. Most importantly, the PRL-3 mAbs were assessed on 282 human colorectal tissue samples (121 normal, 17 adenomas, and 144 adenocarcinomas). PRL-3 protein was detected in 11% of adenocarcinoma samples. The PRL-3- and PRL-1-specific mAbs were further examined on 204 human multiple cancer tissues. The differential expressions of PRL-3 and PRL-1 confirmed the mAbs' specificity. CONCLUSIONS: Using several approaches, we show that PRL-3- or PRL-1-specific mAbs react only to their respective antigen. The expression of PRL-3 in >10% of primary colorectal cancer samples indicates that PRL-3 may prime the metastatic process. These mAbs will be useful as markers in clinical diagnosis for assessing tumor aggressiveness. PMID- 15788668 TI - Accurate discrimination of Barrett's esophagus and esophageal adenocarcinoma using a quantitative three-tiered algorithm and multimarker real-time reverse transcription-PCR. AB - Esophageal adenocarcinoma (EA) is increasing faster than any other cancer in the U.S. In this report, we first show that EA can be distinguished from normal esophagus (NE) and esophageal squamous cell carcinoma by plotting expression values for EpCam, TFF1, and SBEM in three-dimensional Euclidean space. For monitoring progression of Barrett's esophagus (BE) to EA, we developed a highly sensitive assay for limited quantities of tissue whereby 50 ng of RNA are first converted to cDNA using 16 gene-specific primers. Using a set of training tissues, we developed a novel quantitative three-tiered algorithm that allows for accurate (overall accuracy = 61/63, 97%) discrimination of BE versus EA tissues using only three genes. The gene used in the first tier of the algorithm is TSPAN: samples not diagnosed as BE or EA by TSPAN in the first tier are then subjected to a second-tier analysis using ECGF1, followed by a third-tier analysis using SPARC. Addition of TFF1 and SBEM to the first tier (i.e., a five gene marker panel) increases the overall accuracy of the assay to 98% (62/63) and results in mean molecular diagnostic scores (+/- SD) that are significantly different between EA and BE samples (3.19 +/- 1.07 versus -2.74 +/- 1.73, respectively). Our results suggest that relatively few genes can be used to monitor progression of BE to EA. PMID- 15788669 TI - High gene expression of TS1, GSTP1, and ERCC1 are risk factors for survival in patients treated with trimodality therapy for esophageal cancer. AB - PURPOSE: To assess the relationship between molecular markers associated with chemotherapy resistance and survival in esophageal cancer patients treated with trimodality therapy. EXPERIMENTAL DESIGN: The original pretreatment formalin fixed, paraffin-embedded endoscopic esophageal tumor biopsy material was obtained from 99 patients treated with concurrent cisplatin plus 5-fluorouracil plus 45 Gy radiation followed by resection at Duke University Medical Center (Durham, NC) from 1986 to 1997. cDNA was derived from the biopsy and analyzed to determine mRNA expression relative to an internal reference gene (beta-actin) using fluorescence-based, real-time reverse transcription-PCR. Possible markers of platinum chemotherapy association [glutathione S-transferase pi (GSTP1) and excision cross-complementing gene 1 (ERCC1)] and 5-fluorouracil association [thymidylate synthase 1 (TS1)] were measured. RESULTS: Cox proportional hazards model revealed a significant inverse, linear effect for TS1 with respect to survival (P = 0.007). An inverse relationship between TS1 expression and treatment response was also detected (P < or = 0.001). Univariate analysis identified an association with decreased survival for GSTP1 > or = 3.0 (P = 0.05). In multivariate analyses, TS1 >6.0, ERCC1 >3, and GSTP1 >3 were statistically significant predictors of decreased survival (P = 0.007). Additionally, the presence of ERCC1 >3.0 or TS1 >6.0 was associated with an approximately 2-fold increase in the risk of cancer recurrence (P = 0.086 and 0.003, respectively). CONCLUSION: The measurement of relative gene expression of molecular markers associated with chemoresistance in endoscopic esophageal tumor biopsies may be a useful tool in assessing outcome in patients with trimodality treated esophageal cancer. These data should be validated further in larger prospective studies. PMID- 15788670 TI - Prognostic significance of co-expression of RON and MET receptors in node negative breast cancer patients. AB - PURPOSE: RON and MET belong to a subfamily of tyrosine kinase receptors. They both can induce invasive growth, including migration, cell dissociation, and matrix invasion. Cross-linking experiments show that RON and MET form a noncovalent complex on the cell surface and cooperate in intracellular signaling. We wanted to examine the clinical significance of RON and MET expression patterns in node-negative breast cancer. EXPERIMENTAL DESIGN: We studied the protein expressions of RON and MET in five breast cancer cell lines and a homogeneous cohort of 103 T(1-2)N(0)M(0) breast carcinoma patients, including 52 patients with distant metastases and 51 patients with no evidence of disease after at least a 10-year follow-up. RESULTS: Both HCC1937 and MDA-MB-231 cancer cell lines co-overexpressed RON and MET. The MCF-7 cell line did not express RON or MET. In multiple logistic regression analysis, RON expression (odds ratio, 2.6; P = 0.05) and MET expression (odds ratio, 4.7; P = 0.009) were independent predictors of distant relapse. RON+/MET+ and RON-/MET+ tumors resulted in a large risk increase for 10-year disease-free survival after adjusting for tumor size, histologic grade, estrogen receptor, bcl-2, HER-2/neu, and p53 status by multivariate Cox analysis (risk ratio, 5.3; P = 0.001 and risk ratio, 3.76; P = 0.005). The 10 year disease-free survival was 79.3% in patients with RON-/MET- tumors, was only 11.8% in patients with RON+/MET+ tumors, and was 43.9% and 55.6% in patients with RON-/MET+ and RON+/MET- tumors. CONCLUSIONS: Co-expression of RON and MET seems to signify an aggressive phenotype in node-negative breast cancer patients. PMID- 15788671 TI - Signet-ring cell or mucinous histology after preoperative chemoradiation and survival in patients with esophageal or esophagogastric junction adenocarcinoma. AB - PURPOSE: The survival of patients with local-regional adenocarcinoma of the esophagus or esophagogastric junction (EGJ) treated with preoperative chemoradiation is much better in patients with pathologic complete response than those with residual tumor. Some adenocarcinomas have mixed patterns, including signet-ring cell and mucinous histology, but the clinical significance of these subtypes is unknown. EXPERIMENTAL DESIGN: We studied 412 consecutive patients with esophageal or EGJ adenocarcinoma treated with chemoradiation followed by esophagectomy (193 patients) or surgery alone (219 patients). We evaluated signet ring cell and mucinous histology in the resection and pretherapy biopsy specimens and compared clinicopathologic features with overall survival. RESULTS: The fraction of signet-ring cell and mucinous histology was similar in evaluated specimens of patients treated with preoperative chemoradiation or surgery alone (17% and 18%, respectively). The overall survival rate at 5 years of patients treated with preoperative chemoradiation was significantly better if residual signet-ring cell or mucinous histology was present in the esophagectomy specimen (63% versus 28%; P = 0.02). All 13 patients with acellular mucin pools and no residual carcinoma are still alive after an average follow-up time of 36 months. By contrast, in patients treated with surgery alone, overall survival rate was significantly worse if signet-ring cell or mucinous histology was present (14% versus 30%; P = 0.05). In multivariate analysis, overall survival was independently predicted by presence of signet-ring cell or mucinous histology (P = 0.04). CONCLUSIONS: Our study showed that patients with esophageal or EGJ adenocarcinoma who have signet-ring cell or mucinous histology benefited substantially from preoperative chemoradiation and esophagectomy. PMID- 15788672 TI - Reg IV: a promising marker of hormone refractory metastatic prostate cancer. AB - The diagnosis and management of prostate cancer is hampered by the absence of markers capable of identifying patients with metastatic disease. In order to identify potential new markers for prostate cancer, we compared gene expression signatures of matched androgen-dependent and hormone refractory prostate cancer xenografts. One candidate gene overexpressed in a hormone refractory xenograft was homologous to the regenerating protein gene family, a group of secreted proteins expressed in the gastrointestinal tract and overexpressed in inflammatory bowel disease and cancer. This gene, Reg IV, was confirmed to be differentially expressed in the LAPC-9 hormone refractory xenograft. Consistent with its up-regulation in a hormone refractory xenograft, it is expressed in several prostate tumors after neoadjuvant hormone ablation therapy. As predicted by its sequence homology, it is secreted from transiently transfected cells. It is also expressed strongly in a majority of hormone refractory metastases represented on two high-density tissue microarrays. In comparison, it is not expressed by any normal prostate specimens and only at low levels in approximately 40% of primary tumors. These data support Reg IV as a candidate marker for hormone refractory metastatic prostate cancer. PMID- 15788673 TI - Predictors of the response to gefitinib in refractory non-small cell lung cancer. AB - Gefitinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, has a response rate of 10% to 20% in refractory non-small cell lung carcinoma. Although female gender, adenocarcinoma, and never having smoked are possible markers of a favorable response, mutations of the EGFR gene have also been reported to be highly significant predictors of response. Seventy patients with relapsed non-small cell lung carcinoma were enrolled in the Expanded Access Program. After the drug became available commercially, 28 more patients were treated with gefitinib. Response evaluations were feasible in 80 patients. Twenty seven tumor specimens (8 responders and 19 nonresponders) were available for the sequence analysis of the EGFR gene. The response rate was 25% (20/80) and the disease control rate (remission + stable disease) was 47.5% (38/80). The response rate was significantly higher for adenocarcinoma (41.0%) versus non adenocarcinoma (9.8%, P = 0.001), in those who never smoked (58.8%) versus smokers (15.9%, P < 0.001), and in females (42.1%) versus males (19.7%, P = 0.049). A deletion or mutation of the EGFR gene was found in six of eight responders. Remission was noted in all patients with a mutation, whereas the response rate was 9.5% (2/21) in patients without a mutation (P < 0.001). The predictors of response showed significant correlations with survival and time to progression. In a multivariate logistic analysis, the independent predictors of response were smoking history and adenocarcinoma. Given that 9.5% of smokers and 6.7% of those with non-adenocarcinoma showed a mutation of the EGFR gene, the genetic profile may replace those variables as an independent predictor of a response. PMID- 15788674 TI - Stratification of intermediate-risk endometrial cancer patients into groups at high risk or low risk for recurrence based on tumor gene expression profiles. AB - PURPOSE: Endometrial cancers classified as "intermediate risk" based on clinical and/or pathologic features are associated with a 15% to 20% risk of recurrence. Here, we test whether global gene expression profiling can distinguish intermediate-risk tumors into high-risk and low-risk subgroups. EXPERIMENTAL DESIGN: Tumor specimens were obtained from 75 intermediate-risk endometrial cancer patients, 13 who had recurred and 62 who had not recurred with a median follow-up of 24 months. Gene expression profiles were obtained using the Affymetrix U133A GeneChip oligonucleotide microarray. The genes most associated with risk of recurrence were used to create a risk score using a leave-one-out cross-validation method and the univariate Cox proportional hazards regression model. Time to recurrence curves for the high-risk and low-risk subgroups were estimated using the Kaplan-Meier method, and the difference in time to recurrence between these two subgroups was tested using the log-rank test. RESULTS: There was a significant difference in time to recurrence between high-risk and low-risk patients using risk scores as defined above (P = 0.04). The estimated hazard ratio (95% confidence interval) was 3.07 (1.00-9.43). CONCLUSIONS: Patients with intermediate-risk endometrial cancers identified as high-risk for recurrence according to a gene expression-based risk score have a significantly increased risk for recurrence compared with those classified as low risk. These findings suggest that gene expression profiling can potentially contribute to the clinical classification and management of intermediate-risk endometrial cancers. PMID- 15788675 TI - YKL-40 is a differential diagnostic marker for histologic subtypes of high-grade gliomas. AB - PURPOSE AND EXPERIMENTAL DESIGN: In modern neuro-oncology, no variable affects therapeutic decisions and prognostic estimation more than tumor classification. We showed recently that class prediction models, based on gene expression profiles, classify diagnostically challenging malignant gliomas in a manner that better correlates with clinical outcome than standard pathology. In the present study, we used immunohistochemistry to investigate YKL-40 protein expression in independent sets of glioblastomas and anaplastic oligodendrogliomas to determine whether this single marker can aid classification of these high-grade gliomas. RESULTS AND CONCLUSIONS: Glioblastomas show strikingly more YKL-40 expression than anaplastic oligodendrogliomas. Only 2 of 37 glioblastomas showed completely negative YKL-40 staining in both tumor cells and extracellular matrix, whereas 18 of 29 anaplastic oligodendrogliomas were completely negative in non microgemistocytic tumor cells and extracellular matrix. Tumor cell staining intensity was also markedly different: 84% of glioblastomas showed strong staining intensities of 2+ or 3+ whereas 76% of anaplastic oligodendrogliomas either did not stain or stained at only 1+. YKL-40 staining provided a better class distinction of glioblastoma versus anaplastic oligodendroglioma than glial fibrillary acidic protein, the current standard immunohistochemical marker used to distinguish diagnostically challenging gliomas. Moreover, a combination of YKL 40 and glial fibrillary acidic protein immunohistochemistry afforded even greater diagnostic accuracy in anaplastic oligodendrogliomas. PMID- 15788676 TI - Cytoplasmic phospholipase A2 levels correlate with apoptosis in human colon tumorigenesis. AB - Colon cancers often display perturbations in arachidonic acid metabolism, with elevated levels of cyclooxygenase (COX)-2 expression and prostaglandin E(2) (PGE(2)) production frequently observed. Whereas COX-2 and PGE(2) are associated with cancer cell survival and tumor angiogenesis, arachidonic acid itself is a strong apoptotic signal that may facilitate cancer cell death. To further explore how cancer cells exploit the progrowth actions of prostaglandins while suppressing the proapoptotic actions of intracellular arachidonic acid, we determined the cytoplasmic phospholipase A(2) (cPLA(2)) and COX-2 expression levels in a panel of human colon tumors by immunohistochemistry. Although high levels of cPLA(2) and COX-2 expression are predicted to facilitate maximal prostaglandin production, tumors frequently displayed a high-COX-2/low-cPLA(2) phenotype. The least represented phenotype was the high expression of cPLA(2), a characteristic predicted to generate the highest levels of intracellular arachidonic acid. The potential proapoptotic role of cPLA(2) was supported by a higher frequency of terminal deoxynucleotidyl transferase-mediated nick end labeling staining in cPLA(2)-positive tumors. Moreover, analysis of preneoplastic aberrant crypt foci from high-risk patients suggests that acquisition of the high COX-2/low-cPLA(2) phenotype may arise at an early stage of colon carcinogenesis. We additionally inhibited cPLA(2) in HT-29 cells using antisense oligonucleotides. Our results indicate that cPLA(2) plays an important role in tumor necrosis factor alpha-induced apoptosis in human colon cancer cells. Our data further support the model in which colon cancer growth is favored when intracellular arachidonic acid levels are suppressed by inhibition of cPLA(2) or by a high-COX-2/low-cPLA(2) phenotype. PMID- 15788678 TI - Quantitative assessment of telomerase activity and human telomerase reverse transcriptase messenger RNA levels in pancreatic juice samples for the diagnosis of pancreatic cancer. AB - Measurement of telomerase activity is a promising diagnostic tool for pancreatic cancer. Detection of mRNA for human telomerase reverse transcriptase (hTERT), a catalytic subunit of telomerase, is also a diagnostic candidate. In the present study, we developed a telomeric repeat amplification protocol assay with real time PCR and a protocol for quantification of hTERT mRNA with real-time PCR. To evaluate the feasibility of these methods for diagnosis of pancreatic cancer, we measured telomerase activity and hTERT expression in pancreatic cancer cell lines, pancreatic tissues, and pancreatic juice samples from patients with different pancreatic diseases. There were significant correlations between telomerase activity and hTERT expression in cell lines, tissues, and juice samples. The levels of telomerase activity and hTERT expression were significantly higher in tumoral tissues than in nontumoral tissues. In pancreatic juice specimens, some carcinoma samples showed remarkably high expression of hTERT. However, there were no significant differences in hTERT expression between patients with carcinoma and those with benign diseases, although significant differences in telomerase activity were observed. Our present results suggest that the combined assessment of hTERT and telomerase activities in pancreatic juice provides a potent diagnostic method for pancreatic cancer. PMID- 15788677 TI - Stromelysin 3, Ets-1, and vascular endothelial growth factor expression in oral precancerous and cancerous lesions: correlation with microvessel density, progression, and prognosis. AB - PURPOSE: Identification of molecular changes characteristic of development and progression of oral cancer are of paramount importance for effective intervention. Stromelysin 3 (MMP11) is a unique matrix metalloproteinase shown to have dual function during cancer progression. The transcription factor Ets-1 and vascular endothelial growth factor (VEGF) are important proangiogenic factors in cancer. This study was designed to test the hypothesis that concomitant expression of stromelysin 3, Ets-1, and/or VEGF affects the development, progression, and prognosis of oral cancer. PATIENTS AND METHODS: Immunohistochemical analysis of stromelysin 3, Ets-1, VEGF, and platelet/endothelial cell adhesion molecule 1 (a marker for intratumoral microvessel density) was carried out in serial paraffin embedded tissue sections of 220 oral squamous cell carcinomas (OSCC), 90 precancerous lesions (59 hyperplasias and 31 dysplasias), and 81 matched histologically normal oral tissues. RESULTS: Ets-1, VEGF, and stromelysin 3 expression independently correlated with increased intratumoral microvessel density in precancerous lesions (P = 0.05, 0.001, and 0.026, respectively) as well as in SCCs (P = 0.005, 0.01, and 0.031, respectively). Logistic regression analysis revealed that concomitant expression of stromelysin 3 and Ets-1 (stromelysin 3(+)/ Ets-1(+) phenotype; odds ratio, 3.7; P = 0.001) was the most significant predictor for transition to precancerous stage, whereas dual expression of stromelysin 3 and VEGF (stromelysin 3(+)/ VEGF(+) phenotype; odds ratio, 2.07; P = 0.004) was the most important predictor for progression from precancerous stage to frank malignancy. Intriguingly, Ets-1 expression was significantly associated with VEGF expression and stromelysin 3 expression in precancerous tissues as well as OSCCs. Follow-up data for 144 patients for a maximum period of 115 months showed that VEGF [hazards ratio (HR), 4.532; P = 0.004] and Ets-1 (HR = 2.182; P = 0.049) expression significantly correlated with reduced disease-free survival in univariate analysis. In bivariate analysis, patients harboring Ets-1(+)/VEGF(+) phenotype had the worst survival (median disease-free survival, 50 months; HR, 2.943; P = 0.003). Multivariate analysis using Cox's proportional hazards model showed that increased VEGF expression was the most significant adverse prognosticator in OSCC patients (HR, 4.470; P = 0.004). CONCLUSIONS: In conclusion, this study provides the first evidence of concomitant expression of stromelysin 3, VEGF, and Ets-1 in clinical specimens in different stages of development of oral cancer. In early stages, concomitant expression of stromelysin 3 and Ets-1 favors the development of a precancerous state, whereas dual expression of stromelysin 3 and VEGF is associated with progression from precancerous to cancerous state. VEGF expression is an adverse prognosticator for disease-free survival. PMID- 15788679 TI - Legumain expression in relation to clinicopathologic and biological variables in colorectal cancer. AB - PURPOSE: Legumain, a novel asparaginyl endopeptidase, has been observed to be highly expressed in several types of tumors including colorectal cancer. However, there is no study examining the relationship of legumain expression to clinocopathologic and biological variables in colorectal cancers. EXPERIMENTAL DESIGN: We investigated legumain expression in 164 primary colorectal cancers, 34 corresponding distant normal mucosa samples, 89 adjacent normal mucosa samples, and 33 lymph node metastases using immunohistochemistry. We also did Western blotting analysis on three additional colorectal cancers and three colonic cell lines. RESULTS: Legumain expression was increased in primary tumors compared with distant or adjacent normal mucosa (P < 0.05), but there was no significant change between primary tumors and metastases (P > 0.05). Legumain expression was positively related to poorer differentiation/mucinous carcinoma (P = 0.04), higher degree of necrosis (P = 0.03) and apoptosis (P < 0.0001), positive proliferating cell nuclear antigen (P < 0.0001) and p53 expression (P = 0.049), and had a positive tendency towards stromelysin 3 (P = 0.058) and PINCH positivity (P = 0.05). The patients with tumors that showed both weak and lower percentage of the legumain expression, either in tumor (P = 0.01) or in stroma (P = 0.04), had a better prognosis. CONCLUSIONS: The legumain expression may be involved in colorectal cancer development and have a prognostic value in the patients. PMID- 15788680 TI - Gefitinib in patients with malignant mesothelioma: a phase II study by the Cancer and Leukemia Group B. AB - PURPOSE: The Cancer and Leukemia Group B conducted a phase II study of gefitinib, an inhibitor of the epidermal growth factor receptor (EGFR) tyrosine kinase, in patients with previously untreated malignant mesothelioma. EXPERIMENTAL DESIGN: Eligible patients had unresectable pleural or peritoneal mesothelioma, measurable disease, no prior therapy, and performance status 0-1 by Cancer and Leukemia Group B criteria. Gefitinib (500 mg p.o.) was administered once a day for 21 days. Patients underwent restaging after every two cycles. Therapy was continued until disease progression or unacceptable toxicity. RESULTS: The most common grade 3 toxicities were diarrhea (16%) and nausea (12%). Of 43 patients enrolled, 1 patient (2%) had a complete response, 1 patient (2%) had a partial response, 21 (49%) had stable disease lasting two to eight cycles, 15 (35%) had progressive disease, and 5 (12%) had early deaths. One-year survival was 32% [95% confidence interval (CI), 21-50%]. Median survival and failure-free survival were 6.8% (95% CI, 3.5-10.3) and 2.6 months (95% CI, 1.5-4.0), respectively. The 3-month failure free survival was 40% (95% CI, 25-56%). EGFR expression score by immunohistochemistry done in 28 patients was categorized as low (EGFR 1+ or 2+) or high (EGFR 3+) expression: 97% had EGFR overexpression (2+ or 3+). The median and 3-month failure-free survival were 3.6 months and 40% for those patients with low EGFR expression compared with 8.1 and 40% for those with high EGFR expression. CONCLUSIONS: Although 97% of patients with mesothelioma had EGFR overexpression, gefitinib was not active in malignant mesothelioma. EGFR expression does not correlate with failure-free survival. PMID- 15788681 TI - 9-cis-retinoic acid treatment increases serum concentrations of alpha-tocopherol in former smokers. AB - PURPOSE: Low serum concentrations of antioxidants may be associated with an increased risk of cancer. Based on the accumulated evidence, we hypothesized that retinoids would elevate serum alpha-tocopherol. This study was designed to determine whether 9-cis-retinoic acid (9-cis-RA), the most common chemopreventive agent, could alter serum alpha-tocopherol in former smokers. Because hyperlipidemia is a known side effect of retinoids, we also evaluated the association between serum alpha-tocopherol and lipids in the same population. EXPERIMENTAL DESIGN: Subjects who had stopped smoking at least 12 months before the study were randomly assigned to receive oral 9-cis-RA or placebo daily for 3 months. Clinical information and blood samples were obtained monthly; serum alpha tocopherol concentrations were measured by high-performance liquid chromatography and lipid levels by enzymatic assays before treatment and every month during the treatment. RESULTS: Of the 149 subjects in the study, 113 completed 3 months of treatment and provided samples for evaluation of serum alpha-tocopherol. Serum alpha-tocopherol levels in the 9-cis-RA group (n = 52) were higher after treatment (r = 0.445, P < 0.01) than before. The incidences of grade > or =2 hypertriglyceridemia and hypercholesterolemia were higher in the 9-cis-RA group than in the placebo group (P = 0.0005 and P = 0.01, respectively), but there were no serious complications related to hyperlipidemia. CONCLUSIONS: Treatment of former smokers with 9-cis-RA significantly increased their serum alpha-tocopherol levels, and this could be a benefit. In addition, serum alpha-tocopherol could serve as a biomarker for 9-cis-RA treatment. PMID- 15788682 TI - A selective small molecule c-MET Inhibitor, PHA665752, cooperates with rapamycin. AB - PURPOSE: c-MET is believed to be an attractive receptor target for molecular therapeutic inhibition. TPR-MET, a constitutively active oncogenic variant of MET, serves as excellent model for testing c-MET inhibitors. Here, we characterized a small molecule c-MET inhibitor, PHA665752, and tested its cooperation with the mammalian target of rapamycin inhibitor as potential targeted therapy. EXPERIMENTAL DESIGN: The effect of PHA665752 treatment was determined on cell growth, motility and migration, apoptosis, and cell-cycle arrest of TPR-MET-transformed cells. Moreover, the effect of PHA665752 on the phosphorylation on MET, as well as its downstream effectors, p-AKT and p-S6K, was also determined. Finally, growth of TPR-MET-transformed cells was tested in the presence of PHA665752 and rapamycin. H441 non-small cell lung cancer (NSCLC) cells (with activated c-Met) were also tested against both PHA665752 and rapamycin. RESULTS: PHA665752 specifically inhibited cell growth in BaF3. TPR-MET cells (IC(50) < 0.06 micromol/L), induced apoptosis and cell cycle arrest. Constitutive cell motility and migration of the BaF3. TPR-MET cells was also inhibited. PHA665752 inhibited specific phosphorylation of TPR-MET as well as phosphorylation of downstream targets of the mammalian target of rapamycin pathway. When combined with PHA665752, rapamycin showed cooperative inhibition to reduce growth of BaF3. TPR-MET- and c-MET-expressing H441 NSCLC cells. CONCLUSIONS: PHA665752 is a potent small molecule-selective c-MET inhibitor and is highly active against TPR-MET-transformed cells both biologically and biochemically. PHA665752 is also active against H441 NSCLC cells. The c-MET inhibitor can cooperate with rapamycin in therapeutic inhibition of NSCLC, and in vivo studies of this combination against c-MET expressing cancers would be merited. PMID- 15788683 TI - Cyclosporin A is a broad-spectrum multidrug resistance modulator. AB - PURPOSE: Overexpression of the multidrug resistance proteins P-glycoprotein (Pgp), multidrug resistance protein (MRP-1), breast cancer resistance protein (BCRP), and lung resistance protein (LRP) is associated with treatment failure in acute myeloid leukemia (AML) and other malignancies. The Pgp modulator cyclosporin A has shown clinical efficacy in AML, whereas its analogue PSC-833 has not. Cyclosporin A is known to also modulate MRP-1, and we hypothesized that broad-spectrum multidrug resistance modulation might contribute to its clinical efficacy. EXPERIMENTAL DESIGN: We studied the effects of cyclosporin A and PSC 833 on in vitro drug retention and cytotoxicity in resistant cell lines overexpressing Pgp, MRP-1, and BCRP and on nuclear-cytoplasmic drug distribution and cytotoxicity in cells overexpressing LRP. Cellular drug content was assessed by flow cytometry and nuclear-cytoplasmic drug distribution by confocal microscopy. RESULTS: Cyclosporin A enhanced retention of the substrate drug mitoxantrone in cells overexpressing Pgp (HL60/VCR), MRP-1 (HL60/ADR), and BCRP (8226/MR20, HEK-293 482R) and increased cytotoxicity 6-, 4-, 4-, and 3-fold, respectively. Moreover, cyclosporin A enhanced nuclear distribution of doxorubicin in 8226/MR20 cells, which also express LRP, and increased doxorubicin cytotoxicity 12-fold without an effect on cellular doxorubicin content, consistent with expression of wild-type BCRP, which does not efflux doxorubicin. Cyclosporin A also enhanced nuclear doxorubicin distribution in a second cell line with LRP overexpression, HT1080/DR4. PSC-833 enhanced mitoxantrone retention and cytotoxicity in cells overexpressing Pgp, but had no effect in cells overexpressing MRP-1, BCRP, or LRP. CONCLUSIONS: Cyclosporin A modulates Pgp, MRP 1, BCRP, and LRP, and this broad-spectrum activity may contribute to its clinical efficacy. PMID- 15788684 TI - Enhanced natural killer cell binding and activation by low-fucose IgG1 antibody results in potent antibody-dependent cellular cytotoxicity induction at lower antigen density. AB - PURPOSE: Recent studies have revealed that fucose removal from the oligosaccharides of human IgG1 antibodies results in a significant enhancement of antibody-dependent cellular cytotoxicity (ADCC) via improved IgG1 binding to FcgammaRIIIa. In this report, we investigated the relationship between enhanced ADCC and antigen density on target cells using IgG1 antibodies with reduced fucose. EXPERIMENTAL DESIGN: Using EL4 cell-derived transfectants with differential expression levels of exogenous human CC chemokine receptor 4 or human CD20 as target cells, ADCC of fucose variants of chimeric IgG1 antibodies specific for these antigens were measured. We further investigated IgG1 binding to natural killer (NK) cells and NK cell activation during ADCC induction to elucidate the mechanism by which low-fucose IgG1 induces ADCC upon target cells with low antigen expression. RESULTS: Low-fucose IgG1s showed potent ADCC at low antigen densities at which their corresponding high-fucose counterparts could not induce measurable ADCC. The quantitative analysis revealed that fucose depletion could reduce the antigen amount on target cells required for constant degrees of ADCC induction by 10-fold for CC chemokine receptor 4 and 3-fold for CD20. IgG1 binding to NK cells was increased by ligating IgG1 with clustered antigen, especially for low-fucose IgG1. Up-regulation of an activation marker, CD69, on NK cells, particularly the CD56(dim) subset, in the presence of both the antibody and target cells was much greater for the low-fucose antibodies. CONCLUSIONS: Our data showed that fucose removal from IgG1 could reduce the antigen amount required for ADCC induction via efficient recruitment and activation of NK cells. PMID- 15788685 TI - Antiangiogenic treatment with the three thrombospondin-1 type 1 repeats recombinant protein in an orthotopic human pancreatic cancer model. AB - PURPOSE: This study investigates the antiangiogenesis and antitumor efficacy of a recombinant protein composed of the three type 1 repeats (3TSR) of thrombospondin 1 in an orthotopic human pancreatic cancer model and provides useful preclinical data for pancreatic cancer treatment. EXPERIMENTAL DESIGN: Human pancreatic cancer cells (AsPC-1) were injected into the pancreas of severe combined immunodeficient mice. The animals were treated with 3TSR (3 mg per kg per day) or PBS for 3 weeks. Subsequently, the effects of 3TSR on tumor growth, microvessel density, cancer cell proliferation, apoptosis, and endothelial cell apoptosis were analyzed. The in vitro effects of 3TSR on human pancreatic cancer cells were also studied. RESULTS: 3TSR treatment significantly reduced angiogenesis and tumor growth of orthotopic pancreatic cancer. 3TSR-treated mice had a 69% reduction in tumor volume (316.6 +/- 79.3 versus 1,012.2 +/- 364.5 mm(3); P = 0.0001), and a significant increase in tumor necrotic area. After 3TSR treatment, both the vessel number and average microvessel size were significantly decreased, and microvessel density was decreased from 8.0% to 3.7% (P < 0.0001). The apoptotic rate of tumoral endothelial cells in 3TSR-treated tumors increased to 14.7% comparing to 4.2% in control tumors (P < 0.0001). 3TSR showed no direct effects on pancreatic cancer cell proliferation or apoptosis either in vivo or in vitro. CONCLUSION: 3TSR, a domain of a natural occurring angiogenesis inhibitor, showed potent therapeutic effect in pancreatic cancer by inhibiting tumor angiogenesis and may prove to be a promising agent for clinical pancreatic cancer treatment. PMID- 15788686 TI - 4-Hydroxytamoxifen inhibits proliferation of multiple myeloma cells in vitro through down-regulation of c-Myc, up-regulation of p27Kip1, and modulation of Bcl 2 family members. AB - PURPOSE: Multiple myeloma is an incurable B-cell malignancy requiring new therapeutic strategies. Our approach was to analyze the in vitro effects of a selective estrogen receptor modulator, 4-hydroxytamoxifen (4-OHT), on six multiple myeloma cell lines. EXPERIMENTAL DESIGN: Cultured multiple myeloma cells were treated with various 4-OHT concentrations and the cellular response was studied: cell proliferation, cell viability, induction of apoptosis, caspase activities, and expression of signaling proteins. RESULTS: We found that pharmacologic concentrations of 4-OHT inhibit cell proliferation (4 of 6 cell lines). This inhibition is achieved by two independent events: a block at the G(1) phase of the cell cycle and the induction of apoptotic death. The cellular response to 4-OHT depends on the presence of functional estrogen receptors. 4-OHT treatment activates an intrinsic mitochondrial caspase-9-dependent pathway but not the Fas/FasL death pathway. Signaling pathways known to be involved in the survival and/or proliferation of multiple myeloma cells are not affected by 4-OHT treatment. 4-OHT-induced G(1) arrest is accompanied by the up-regulation of the cell cycle inhibitor p27(Kip1) and the down-regulation of c-Myc. Among the Bcl-2 family members tested, the proapoptotic BimS protein is induced whereas the antiapoptotic protein Mcl-1 is decreased. CONCLUSIONS: Although the effects of 4 OHT are observed at micromolar concentrations, cellular mechanisms responsible for G(1) arrest, as well as apoptosis induction, are similar to those observed in breast cancer cells. Our data support the concept that 4-OHT may represent an alternative approach to inhibit proliferation and induce apoptosis of multiple myeloma cells. PMID- 15788687 TI - Casein kinase 2 regulates both apoptosis and the cell cycle following DNA damage induced by 6-thioguanine. AB - PURPOSE: The purine antimetabolite, 6-thioguanine (6-TG), is an effective drug in the management of acute leukemias. In this study, we analyze the mechanisms of apoptosis associated with 6-TG treatment and casein kinase 2 (CK2 or CKII) in human tumor cells. EXPERIMENTAL DESIGN: Small interfering RNA and chemical CK2 inhibitors were used to reduce CK2 activity. Control and CK2 activity-reduced cells were cultured with 6-TG and assessed by flow cytometry to measure apoptosis and cell cycle profiles. Additionally, confocal microscopy was used to assess localization of CK2 catalytic units following 6-TG treatment. RESULTS: Transfection of small interfering RNA against the CK2 alpha and/or alpha' catalytic subunits results in marked apoptosis of HeLa cells following treatment with 6-TG. Chemical inhibitors of CK2 also induce apoptosis following 6-TG treatment. Apoptosis induced by 6-TG is similarly observed in both mismatch repair-proficient and -deficient HCT116 and HeLa cells. Concomitant treatment with a pan-caspase inhibitor or transfection of apoptosis repressor with caspase recruitment domain markedly suppresses the apoptotic response to DNA damage by 6 TG in the CK2-reduced cells, indicating caspase regulation by CK2. CK2 alpha relocalizes to the endoplasmic reticulum after 6-TG treatment. Additionally, transfection of Cdc2 with a mutation at Ser(39) to Ala, which is the CK2 phosphorylation site, partially inhibits cell cycle progression in G(1) to G(2) phase following 6-TG treatment. CONCLUSION: CK2 is essential for apoptosis inhibition following DNA damage induced by 6-TG, controlling caspase activity. PMID- 15788688 TI - Vascular endothelial growth factor: a therapeutic target for tumors of the Ewing's sarcoma family. AB - PURPOSE: We have reported previously that intratumoral microvessel density (MVD) is a significant prognostic indicator of event-free survival in the Ewing's sarcoma family of tumors (ESFT). Here, the angiogenic growth factor expression profile and its relationship with MVD has been investigated in ESFT. EXPERIMENTAL DESIGN AND RESULTS: Using ESFT model systems, the potential of these factors as therapeutic targets has been evaluated. A significant correlation (P = 0.02) was observed between vascular endothelial growth factor (VEGF) expression and MVD, consistent with the hypothesis that VEGF regulates the development of microvessels in ESFT. There was no correlation between MVD and any of the other growth factors studied. All six ESFT cell lines studied produced and secreted VEGF; five of six cell lines also secreted placental growth factor, one cell line (A673) at high levels. Tumor conditioned medium induced proliferation of human umbilical vein endothelial cells. Expression of VEGF receptors Flt-1 and Flk 1/KDR was heterogeneous across the cell lines. Both receptor tyrosine kinase inhibitors SU6668 (targets Flk-1/KDR, platelet-derived growth factor receptor beta, and fibroblast growth factor receptor 1) and SU5416 (targets Flk-1/KDR) as well as anti-VEGF agents rhuMAb-VEGF (bevacizumab) and VEGF Trap delayed s.c. growth of ESFT in mice compared with untreated groups: SU6668 (100 mg/kg/d), SU5416 (25 mg/kg/d), rhuMAb-VEGF (10 mg/kg twice weekly), and VEGF Trap (2.5 or 25 mg/kg twice weekly). CONCLUSIONS: These data suggest that VEGF is the single most important regulator of angiogenesis in ESFT and may be exploited for therapeutic advantage. PMID- 15788689 TI - Methylseleninic acid potentiates apoptosis induced by chemotherapeutic drugs in androgen-independent prostate cancer cells. AB - PURPOSE: To test whether and how selenium enhances the apoptosis potency of selected chemotherapeutic drugs in prostate cancer (PCA) cells. EXPERIMENTAL DESIGN: DU145 and PC3 human androgen-independent PCA cells were exposed to minimal apoptotic doses of selenium and/or the topoisomerase I inhibitor 7-ethyl 10-hydroxycamptothecin (SN38), the topoisomerase II inhibitor etoposide or the microtubule inhibitor paclitaxel/taxol. Apoptosis was measured by ELISA for histone-associated DNA fragments, by flow cytometric analysis of sub-G(1) fraction, and by immunoblot analysis of cleaved poly(ADP-ribose)polymerase. Pharmacologic inhibitors were used to manipulate caspases and c-Jun-NH(2) terminal kinases (JNK). RESULTS: The methylselenol precursor methylseleninic acid (MSeA) increased the apoptosis potency of SN38, etoposide, or paclitaxel by several folds higher than the expected sum of the apoptosis induced by MSeA and each drug alone. The combination treatment did not further enhance JNK1/2 phosphorylation that was induced by each drug in DU145 cells. The JNK inhibitor SP600125 substantially decreased the activation of caspases and apoptosis induced by MSeA combination with SN38 or etoposide and completely blocked these events induced by MSeA/paclitaxel. The caspase-8 inhibitor zIETDfmk completely abolished apoptosis and caspase-9 and caspase-3 cleavage, whereas the caspase-9 inhibitor zLEHDfmk significantly decreased caspase-3 cleavage and apoptosis but had no effect on caspase-8 cleavage. None of these caspase inhibitors abolished JNK1/2 phosphorylation. A JNK-independent suppression of survivin by SN38 and etoposide, but not by paclitaxel, was also observed. In contrast to MSeA, selenite did not show any enhancing effect on the apoptosis induced by these drugs. CONCLUSIONS: MSeA enhanced apoptosis induced by cancer therapeutic drugs in androgen independent PCA cells. In DU145 cells, the enhancing effect was primarily through interactions between MSeA and JNK-dependent targets to amplify the caspase-8 initiated activation cascades. The results suggest a novel use of methyl selenium for improving the chemotherapy of PCA. PMID- 15788690 TI - Elevated physiologic tumor pressure promotes proliferation and chemosensitivity in human osteosarcoma. AB - PURPOSE: This study investigates the effect of constitutively raised interstitial fluid pressure on osteosarcoma physiology and chemosensitivity. EXPERIMENTAL DESIGN: We did pressure and blood flow assessments at the time of open biopsy in patients with the diagnosis of high-grade osteosarcoma and correlated this to survival and chemotherapy-associated tumor necrosis. Osteosarcoma cell lines were then evaluated for proliferative and therapeutic indices in a replicated high pressure environment. RESULTS: Sixteen osteosarcomas in vivo were assessed and exhibited elevated interstitial fluid pressures (mean 35.2 +/- SD, 18.6 mmHg). This was not associated with significantly impeded blood flow as measured by a Doppler probe at a single site (P < 0.12). Nonetheless, greater chemotherapy associated necrosis and associated longer survival were seen in tumors with higher interstitial fluid pressures (P < 0.05). In vitro, cells undergo significant physiologic changes under pressure. Osteosarcoma cell lines grown in a novel hydrostatically pressurized system had variable cell line-specific growth proportional to the level of pressure. They were more proliferative as indicated by cell cycle analysis with more cells in S phase after 48 hours of pressurization (P < 0.01). There was a significant elevation in the cell cycle related transcription factors E2F-1 (P < 0.03) and E2F-4 (P < 0.002). These changes were associated with increased chemosensitivity. Cells tested under pressure showed an increased sensitivity to cisplatin (P < 0.00006) and doxorubicin (P < 0.03) reminiscent of the increased chemotherapy-associated necrosis seen in tumors with higher interstitial fluid pressure in the clinical study. CONCLUSIONS: The results of this study suggest that cells in the in vivo pressurized environment are at a higher state of regenerative activity than is demonstrable in conventional cell culture systems. Variations in tumor interstitial fluid pressure have the potential to alter chemotherapeutic effects. PMID- 15788692 TI - Effective inhibition of experimental human ovarian cancers with a targeted cytotoxic bombesin analogue AN-215. AB - PURPOSE: To determine whether the cytotoxic analogue of bombesin/gastrin releasing peptide (GRP) AN-215 can inhibit the in vivo growth of four human ovarian cancer cell lines. AN-215 consists of 2-pyrrolinodoxorubicin (AN-201), a superactive derivative of doxorubicin linked to a bombesin antagonist carrier des D-Tpi-RC-3095. This conjugate binds strongly to receptors for bombesin/GRP and can be targeted to tumors that express these receptors. Bombesin/GRP receptors are found in 77% of human ovarian cancer specimens. EXPERIMENTAL DESIGN: Nude mice bearing xenografts of ES-2, SKOV-3, OV-1063, and UCI-107 human ovarian carcinomas were treated with AN-215. The antitumor effects and the toxicity were determined. The expression of bombesin receptor subtypes was measured by reverse transcriptase PCR analysis, and the presence of bombesin/GRP receptors was determined by radioligand binding assays. RESULTS: AN-215 significantly (P < 0.05) inhibited growth of ES-2, OV-1063, and UCI-107 tumors, prevented the metastatic spread of ES-2 cancers, and prolonged the survival of nude mice bearing i.p. ES-2 xenografts. Cytotoxic radical AN-201, the unconjugated mixture of bombesin antagonist RC-3095 and AN-201 or RC-3095 alone had no significant effects. Blockade of bombesin/GRP receptors abolished the effect of AN-215. The expression of bombesin/GRP receptors was not changed after repeated treatment with AN-215. CONCLUSIONS: Our findings indicate that targeted chemotherapy with cytotoxic bombesin/GRP analogue AN-215 can inhibit ovarian tumors, which express bombesin/GRP receptors. AN-215 might provide a new treatment modality for women with advanced ovarian carcinoma. PMID- 15788691 TI - Targeting the mevalonate pathway inhibits the function of the epidermal growth factor receptor. AB - PURPOSE: The epidermal growth factor receptor (EGFR) is a key regulator of growth, differentiation, and survival of epithelial cancers. In a small subset of tumors, the presence of activating mutations within the ATP binding site confers increased susceptibility to gefitinib, a potent tyrosine kinase inhibitor of EGFR. Agents that can inhibit EGFR function through different mechanisms may enhance gefitinib activity in patients lacking these mutations. Mevalonate metabolites play significant roles in the function of the EGFR; therefore, mevalonate pathway inhibitors may potentiate EGFR-targeted therapies. EXPERIMENTAL DESIGN: In this study, we evaluated the effect of lovastatin on EGFR function and on gefitinib activity. Effects on EGFR function were analyzed by Western blot analysis using phosphospecific antibodies to EGFR, AKT, and extracellular signal-regulated kinase. Cytotoxic effects of lovastatin and/or gefitinib were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry. RESULTS: Lovastatin treatment inhibited EGF induced EGFR autophosphorylation by 24 hours that was reversed by the coadministration of mevalonate. Combining lovastatin and gefitinib treatments showed enhanced inhibition of AKT activation by EGF in SCC9 cells. The combination of 10 mumol/L lovastatin and 10 mumol/L gefitinib treatments showed cooperative cytotoxicity in all 8 squamous cell carcinomas, 4 of 4 non-small cell lung carcinoma and 4 of 4 colon carcinoma cell lines tested. Isobologram and flow cytometric analyses of three representative cell lines with wild-type EGFR ATP binding sites confirmed that this combination was synergistic inducing a potent apoptotic response. CONCLUSIONS: Taken together, these results show that targeting the mevalonate pathway can inhibit EGFR function. They also suggest the potential utility of combining these clinically relevant therapeutic approaches. PMID- 15788693 TI - Induction of an antigen cascade by diversified subcutaneous/intratumoral vaccination is associated with antitumor responses. AB - PURPOSE: Cancer vaccines targeting tumor-associated antigens are being investigated for the therapy of tumors. Numerous strategies, including the direct intratumoral (i.t.) vaccination route, have been examined. For tumors expressing carcinoembryonic antigen (CEA) as a model tumor-associated antigen, we previously designed poxviral vectors that contain the transgenes for CEA and a triad of T cell costimulatory molecules, B7-1, intercellular adhesion molecule-1, (ICAM-1), and leukocyte function associated antigen-3 (LFA-3) (CEA/TRICOM). Two types of poxvirus vectors were developed: replication-competent recombinant vaccinia and replication-defective recombinant fowlpox. We have shown previously that a vaccine regimen composed of priming mice s.c. with recombinant vaccinia CEA/TRICOM and boosting i.t. with recombinant fowlpox-CEA/TRICOM was superior to priming and boosting vaccinations using the conventional s.c. route in inducing T cell responses specific for CEA. These studies also showed that CEA was needed to be present both in the vaccine and in the tumor for therapeutic effects. EXPERIMENTAL DESIGN: To determine specific immune responses associated with vaccination-mediated tumor regression, CEA-transgenic mice bearing CEA(+) tumors were vaccinated with the CEA/TRICOM s.c./i.t. regimen, and T-cell immune responses were assessed. RESULTS: In CEA(+) tumor-bearing mice vaccinated with the CEA/TRICOM s.c./i.t. regimen, T-cell responses could be detected not only to CEA encoded in vaccine vectors but also to other antigens expressed on the tumor itself: wild-type p53 and an endogenous retroviral epitope of gp70. Moreover, the magnitude of CD8(+) T-cell immune responses to gp70 was far greater than that induced to CEA or p53. Finally, the predominant T-cell population infiltrating the regressing CEA(+) tumor after therapy was specific for gp70. CONCLUSION: These studies show that the breadth and magnitude of antitumor immune cascades to multiple antigens could be critical in the therapy of established tumors. PMID- 15788694 TI - Production of thymus and activation-regulated chemokine and macrophage-derived chemokine by CCR4+ adult T-cell leukemia cells. AB - PURPOSE: Adult T-cell leukemia/lymphoma (ATL) is a peripheral CD4(+)CD25(+) T cell malignancy caused by human T-cell leukemia virus type I. The tumor cells frequently infiltrate in the skin, lymph nodes and other organs and especially form prominent cutaneous masses. Recently, ATL cells have been shown to express Th2 chemokine receptor CCR4. The aim of this study is to investigate the possibility that CCR4 ligands, thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC), are produced by CCR4(+) ATL cells per se. EXPERIMENTAL DESIGN: CD4(+) or CD4(+)CD14(-) cells were purified from peripheral blood mononuclear cells of 11 ATL patients with cutaneous involvement and normal healthy volunteers. Tissue-infiltrating cells were isolated from skin tumors. The expression of chemokine receptors on these cells were analyzed by flow cytometry. The production of chemokines and cytokines by the neoplastic cells was assessed by ELISA and reverse transcription-PCR after cultivation for 96 hours in the presence or absence of anti-CD3/CD28 monoclonal antibodies. Finally, TARC and CCR4 expressions were examined by immunohistochemistry. RESULTS: ATL cells highly expressed CCR4 but did not necessarily exhibit the Th2 cytokine profile. The cells also produced TARC and MDC. The production level of MDC was higher in the skin tumor formation group than that in the nontumor group. Immunohistochemically, both CCR4 and TARC were expressed by the tumor cells in the lesional skin. CONCLUSIONS: ATL cells not only express CCR4 but also produce TARC and MDC. The skin tumor formation as well as the monoclonal integration of proviral DNA are the factors that are associated with the high production of Th2 chemokines by ATL cells. PMID- 15788695 TI - Breast cancer resistance protein in drug resistance of primitive CD34+38- cells in acute myeloid leukemia. AB - PURPOSE: Acute myeloid leukemia (AML) is considered a stem cell disease. Incomplete chemotherapeutic eradication of leukemic CD34+38- stem cells is likely to result in disease relapse. The purpose of this study was to investigate the role of the breast cancer resistance protein (BCRP/ATP-binding cassette, subfamily G, member 2) in drug resistance of leukemic stem cells and the effect of its modulation on stem cell eradication in AML. EXPERIMENTAL DESIGN: BCRP expression (measured flow-cytometrically using the BXP21 monoclonal antibody) and the effect of its modulation (using the novel fumitremorgin C analogue KO143) on intracellular mitoxantrone accumulation and in vitro chemosensitivity were assessed in leukemic CD34+38- cells. RESULTS: BCRP was preferentially expressed in leukemic CD34+38- cells and blockage of BCRP-mediated drug extrusion by the novel fumitremorgin C analogue KO143 resulted in increased intracellular mitoxantrone accumulation in these cells in the majority of patients. This increase, however, was much lower than in the mitoxantrone-resistant breast cancer cell line MCF7-MR and significant drug extrusion occurred in the presence of BCRP blockage due to the presence of additional drug transport mechanisms, among which ABCB1 and multiple drug resistance protein. In line with these findings, selective blockage of BCRP by KO143 did not enhance in vitro chemosensitivity of leukemic CD34+38- cells. CONCLUSIONS: These results show that drug extrusion from leukemic stem cells is mediated by the promiscuous action of BCRP and additional transporters. Broad-spectrum inhibition, rather than modulation of single mechanisms, is therefore likely to be required to circumvent drug resistance and eradicate leukemic stem cells in AML. PMID- 15788697 TI - Do drugs that stimulate ovulation increase the risk for endometrial stromal sarcoma? PMID- 15788696 TI - Coxsackievirus and adenovirus receptor expression on primary osteosarcoma specimens and implications for gene therapy with recombinant adenoviruses. PMID- 15788698 TI - Hysterectomy and bilateral oophorectomy for severe premenstrual syndrome. PMID- 15788699 TI - Ovarian reserve and reproductive age may be determined from measurement of ovarian volume by transvaginal sonography. PMID- 15788700 TI - Augmented prefrontal acetylcholine release during challenged attentional performance. AB - Previous research has demonstrated that attentional performance depends on the integrity of the cortical cholinergic input system and that such performance is associated with increases in cortical acetylcholine (ACh) release. The present experiment tested the hypothesis that the attentional impairments produced by bilateral basal forebrain infusions of the NMDA receptor antagonist DL-2-amino-5 phosphonovaleric acid (APV) are associated with attenuation of performance associated increases in ACh release. Rats were trained in a sustained attention task and equipped with three guide cannula for the bilateral infusion of the NMDA receptor antagonist APV (0, 3, 20 nmol) and for the insertion of a dialysis probe into the medial prefrontal cortex (mPFC). APV or vehicle was infused remotely following completion of the first of five blocks of trials. During the first block, attentional performance was associated with a 140% increase in ACh efflux. Infusions of APV decreased the animals' ability to detect signals and augmented the increases in ACh efflux observed prior to infusions. These data indicate a dissociation between levels of attentional performance and increases in mPFC ACh release. Augmentation of performance-associated increases in mPFC cholinergic transmission is hypothesized to mediate the increased demands on attentional 'effort' that are required to maintain performance under challenging conditions. PMID- 15788701 TI - Prenatal exposure to the CB1 receptor agonist WIN 55,212-2 causes learning disruption associated with impaired cortical NMDA receptor function and emotional reactivity changes in rat offspring. AB - The aim of this study was to investigate whether prenatal exposure to the cannabinoid CB1 receptor agonist WIN 55,212-2 (WIN) at a daily dose devoid of overt signs of toxicity and/or gross malformations (0.5 mg/kg, gestation days 5 20), influences cortical glutamatergic neurotransmission, learning and emotional reactivity in rat offspring. Basal and K+-evoked extracellular glutamate levels were significantly lower in cortical cell cultures obtained from pups exposed to WIN during gestation with respect to those measured in cultures obtained from neonates born from vehicle-treated dams. The addition of NMDA to cortical cell cultures from neonates born from vehicle-treated dams concentration-dependently increased glutamate levels, and this was absent in cell cultures obtained from WIN-exposed pups. WIN-exposed rats also revealed a poorer performance in homing (10-12 days of age) and active avoidance tests (80 days of age) as well as a decrease in the rate of separation-induced ultrasonic emission (10 days of age). Finally, prenatal exposure to WIN induced a reduction in the number of cortical neuronal population. These findings (i) provide evidence for a deficit in cortical glutamatergic neurotransmission and behaviour in the rat neonate following prenatal exposure to WIN; and (ii) suggest that the reduction in cortical glutamatergic neurotransmission, NMDA receptor activity and alterations in neuronal development might underlie, at least in part, the learning deficit and decreased emotional reactivity observed in the offspring. PMID- 15788702 TI - Frontal lobe mechanisms that resolve proactive interference. AB - Memory of a past experience can interfere with processing during a subsequent experience, a phenomenon termed proactive interference (PI). Neuroimaging and neuropsychological evidence implicate the left mid-ventrolateral prefrontal cortex (mid-VLPFC) in PI resolution during short-term item recognition, though the precise mechanisms await specification. The present functional magnetic resonance imaging (fMRI) experiment sought to further constrain theorizing regarding PI resolution. On each trial, subjects maintained a target set of words, and then decided if a subsequent probe was contained in the target set (positive) or not (negative). Importantly, for half of the negative and half of the positive trials, the probe had been contained in the previous target set (recent). Relative to non-recent trials, negative-recent trials produced an increase in response times and error rates, behavioral markers of PI. In fMRI measures, negative recency was associated with increased activation in the left mid-VLPFC, as well as in the bilateral fronto-polar cortex, providing evidence for multiple components in PI resolution. Furthermore, recency effects were evident during both negative and positive trials, with the magnitude of the recency effect in the mid-VLPFC being greater on negative trials. Collectively, these results serve to specify and constrain proposed models of PI resolution. PMID- 15788703 TI - Molecular physiology and pathophysiology of electroneutral cation-chloride cotransporters. AB - Electroneutral cation-Cl(-) cotransporters compose a family of solute carriers in which cation (Na(+) or K(+)) movement through the plasma membrane is always accompanied by Cl(-) in a 1:1 stoichiometry. Seven well-characterized members include one gene encoding the thiazide-sensitive Na(+)-Cl(-) cotransporter, two genes encoding loop diuretic-sensitive Na(+)-K(+)-2Cl(-) cotransporters, and four genes encoding K(+)-Cl(-) cotransporters. These membrane proteins are involved in several physiological activities including transepithelial ion absorption and secretion, cell volume regulation, and setting intracellular Cl(-) concentration below or above its electrochemical potential equilibrium. In addition, members of this family play an important role in cardiovascular and neuronal pharmacology and pathophysiology. Some of these cotransporters serve as targets for loop diuretics and thiazide-type diuretics, which are among the most commonly prescribed drugs in the world, and inactivating mutations of three members of the family cause inherited diseases such as Bartter's, Gitelman's, and Anderman's diseases. Major advances have been made in the past decade as consequences of molecular identification of all members in this family. This work is a comprehensive review of the knowledge that has evolved in this area and includes molecular biology of each gene, functional properties of identified cotransporters, structure-function relationships, and physiological and pathophysiological roles of each cotransporter. PMID- 15788704 TI - Ghrelin: structure and function. AB - Small synthetic molecules called growth hormone secretagogues (GHSs) stimulate the release of growth hormone (GH) from the pituitary. They act through the GHS R, a G protein-coupled receptor whose ligand has only been discovered recently. Using a reverse pharmacology paradigm with a stable cell line expressing GHS-R, we purified an endogenous ligand for GHS-R from rat stomach and named it "ghrelin," after a word root ("ghre") in Proto-Indo-European languages meaning "grow." Ghrelin is a peptide hormone in which the third amino acid, usually a serine but in some species a threonine, is modified by a fatty acid; this modification is essential for ghrelin's activity. The discovery of ghrelin indicates that the release of GH from the pituitary might be regulated not only by hypothalamic GH-releasing hormone, but also by ghrelin derived from the stomach. In addition, ghrelin stimulates appetite by acting on the hypothalamic arcuate nucleus, a region known to control food intake. Ghrelin is orexigenic; it is secreted from the stomach and circulates in the bloodstream under fasting conditions, indicating that it transmits a hunger signal from the periphery to the central nervous system. Taking into account all these activities, ghrelin plays important roles for maintaining GH release and energy homeostasis in vertebrates. PMID- 15788705 TI - Adult neurogenesis: from precursors to network and physiology. AB - The discovery that the adult mammalian brain creates new neurons from pools of stemlike cells was a breakthrough in neuroscience. Interestingly, this particular new form of structural brain plasticity seems specific to discrete brain regions, and most investigations concern the subventricular zone (SVZ) and the dentate gyrus (DG) of the hippocampal formation (HF). Overall, two main lines of research have emerged over the last two decades: the first aims to understand the fundamental biological properties of neural stemlike cells (and their progeny) and the integration of the newly born neurons into preexisting networks, while the second focuses on understanding its relevance in brain functioning, which has been more extensively approached in the DG. Here, we propose an overview of the current knowledge on adult neurogenesis and its functional relevance for the adult brain. We first present an analysis of the methodological issues that have hampered progress in this field and describe the main neurogenic sites with their specificities. We will see that despite considerable progress, the levels of anatomic and functional integration of the newly born neurons within the host circuitry have yet to be elucidated. Then the intracellular mechanisms controlling neuronal fate are presented briefly, along with the extrinsic factors that regulate adult neurogenesis. We will see that a growing list of epigenetic factors that display a specificity of action depending on the neurogenic site under consideration has been identified. Finally, we review the progress accomplished in implicating neurogenesis in hippocampal functioning under physiological conditions and in the development of hippocampal-related pathologies such as epilepsy, mood disorders, and addiction. This constitutes a necessary step in promoting the development of therapeutic strategies. PMID- 15788706 TI - Developmental origins of the metabolic syndrome: prediction, plasticity, and programming. AB - The "fetal" or "early" origins of adult disease hypothesis was originally put forward by David Barker and colleagues and stated that environmental factors, particularly nutrition, act in early life to program the risks for adverse health outcomes in adult life. This hypothesis has been supported by a worldwide series of epidemiological studies that have provided evidence for the association between the perturbation of the early nutritional environment and the major risk factors (hypertension, insulin resistance, and obesity) for cardiovascular disease, diabetes, and the metabolic syndrome in adult life. It is also clear from experimental studies that a range of molecular, cellular, metabolic, neuroendocrine, and physiological adaptations to changes in the early nutritional environment result in a permanent alteration of the developmental pattern of cellular proliferation and differentiation in key tissue and organ systems that result in pathological consequences in adult life. This review focuses on those experimental studies that have investigated the critical windows during which perturbations of the intrauterine environment have major effects, the nature of the epigenetic, structural, and functional adaptive responses which result in a permanent programming of cardiovascular and metabolic function, and the role of the interaction between the pre- and postnatal environment in determining final health outcomes. PMID- 15788707 TI - Embryonic stem cells: prospects for developmental biology and cell therapy. AB - Stem cells represent natural units of embryonic development and tissue regeneration. Embryonic stem (ES) cells, in particular, possess a nearly unlimited self-renewal capacity and developmental potential to differentiate into virtually any cell type of an organism. Mouse ES cells, which are established as permanent cell lines from early embryos, can be regarded as a versatile biological system that has led to major advances in cell and developmental biology. Human ES cell lines, which have recently been derived, may additionally serve as an unlimited source of cells for regenerative medicine. Before therapeutic applications can be realized, important problems must be resolved. Ethical issues surround the derivation of human ES cells from in vitro fertilized blastocysts. Current techniques for directed differentiation into somatic cell populations remain inefficient and yield heterogeneous cell populations. Transplanted ES cell progeny may not function normally in organs, might retain tumorigenic potential, and could be rejected immunologically. The number of human ES cell lines available for research may also be insufficient to adequately determine their therapeutic potential. Recent molecular and cellular advances with mouse ES cells, however, portend the successful use of these cells in therapeutics. This review therefore focuses both on mouse and human ES cells with respect to in vitro propagation and differentiation as well as their use in basic cell and developmental biology and toxicology and presents prospects for human ES cells in tissue regeneration and transplantation. PMID- 15788708 TI - Links between dietary salt intake, renal salt handling, blood pressure, and cardiovascular diseases. AB - Epidemiological, migration, intervention, and genetic studies in humans and animals provide very strong evidence of a causal link between high salt intake and high blood pressure. The mechanisms by which dietary salt increases arterial pressure are not fully understood, but they seem related to the inability of the kidneys to excrete large amounts of salt. From an evolutionary viewpoint, the human species is adapted to ingest and excrete <1 g of salt per day, at least 10 times less than the average values currently observed in industrialized and urbanized countries. Independent of the rise in blood pressure, dietary salt also increases cardiac left ventricular mass, arterial thickness and stiffness, the incidence of strokes, and the severity of cardiac failure. Thus chronic exposure to a high-salt diet appears to be a major factor involved in the frequent occurrence of hypertension and cardiovascular diseases in human populations. PMID- 15788709 TI - Respiratory action of the intercostal muscles. AB - The mechanical advantages of the external and internal intercostals depend partly on the orientation of the muscle but mostly on interspace number and the position of the muscle within each interspace. Thus the external intercostals in the dorsal portion of the rostral interspaces have a large inspiratory mechanical advantage, but this advantage decreases ventrally and caudally such that in the ventral portion of the caudal interspaces, it is reversed into an expiratory mechanical advantage. The internal interosseous intercostals in the caudal interspaces also have a large expiratory mechanical advantage, but this advantage decreases cranially and, for the upper interspaces, ventrally as well. The intercartilaginous portion of the internal intercostals (the so-called parasternal intercostals), therefore, has an inspiratory mechanical advantage, whereas the triangularis sterni has a large expiratory mechanical advantage. These rostrocaudal gradients result from the nonuniform coupling between rib displacement and lung expansion, and the dorsoventral gradients result from the three-dimensional configuration of the rib cage. Such topographic differences in mechanical advantage imply that the functions of the muscles during breathing are largely determined by the topographic distributions of neural drive. The distributions of inspiratory and expiratory activity among the muscles are strikingly similar to the distributions of inspiratory and expiratory mechanical advantages, respectively. As a result, the external intercostals and the parasternal intercostals have an inspiratory function during breathing, whereas the internal interosseous intercostals and the triangularis sterni have an expiratory function. PMID- 15788710 TI - Store-operated calcium channels. AB - In electrically nonexcitable cells, Ca(2+) influx is essential for regulating a host of kinetically distinct processes involving exocytosis, enzyme control, gene regulation, cell growth and proliferation, and apoptosis. The major Ca(2+) entry pathway in these cells is the store-operated one, in which the emptying of intracellular Ca(2+) stores activates Ca(2+) influx (store-operated Ca(2+) entry, or capacitative Ca(2+) entry). Several biophysically distinct store-operated currents have been reported, but the best characterized is the Ca(2+) release activated Ca(2+) current, I(CRAC). Although it was initially considered to function only in nonexcitable cells, growing evidence now points towards a central role for I(CRAC)-like currents in excitable cells too. In spite of intense research, the signal that relays the store Ca(2+) content to CRAC channels in the plasma membrane, as well as the molecular identity of the Ca(2+) sensor within the stores, remains elusive. Resolution of these issues would be greatly helped by the identification of the CRAC channel gene. In some systems, evidence suggests that store-operated channels might be related to TRP homologs, although no consensus has yet been reached. Better understood are mechanisms that inactivate store-operated entry and hence control the overall duration of Ca(2+) entry. Recent work has revealed a central role for mitochondria in the regulation of I(CRAC), and this is particularly prominent under physiological conditions. I(CRAC) therefore represents a dynamic interplay between endoplasmic reticulum, mitochondria, and plasma membrane. In this review, we describe the key electrophysiological features of I(CRAC) and other store-operated Ca(2+) currents and how they are regulated, and we consider recent advances that have shed insight into the molecular mechanisms involved in this ubiquitous and vital Ca(2+) entry pathway. PMID- 15788711 TI - The effect of emotion and personality on olfactory perception. AB - It is well established that both the emotional tone of sensory stimuli and the personality characteristics of an individual can bias sensory perception. What has largely been unexplored is whether the current emotional state of an individual has a similar effect, and how it works together with other factors. Here we carry out a comprehensive study to examine how olfactory perception is affected by the emotional tone of the stimuli, and the personality and current emotional state of the individual. Subjects reported experiencing happiness, sadness, negativity/hostility and neutrality when exposed to corresponding emotionally themed video clips, and in each case, smelled a suprathreshold pleasant, an unpleasant and a neutral odorant. The time taken for the subject to detect each odorant and the olfactory intensity were recorded. We found that women detected the pleasant odorant faster than the neutral one. In addition, personality modulated reaction time and olfactory intensity, such that neurotic and anxious individuals were selectively biased toward affective rather than neutral odorants. Finally, current emotional state augmented intensity in men but not in women, and differentially influenced the response time. These findings provided new insights into the effects of emotion and personality on olfactory perception. PMID- 15788712 TI - The significance of background odour for an egg parasitoid to detect plants with host eggs. AB - Scots pine has been shown to produce a volatile bouquet that attracts egg parasitoids in response to oviposition of the herbivorous sawfly Diprion pini. Previous analyses of headspace volatiles of oviposition-induced pine twigs revealed only quantitative changes; in particular, the sesquiterpene (E)-beta farnesene was emitted in significantly higher quantities by oviposition-induced pine. Here we investigated whether (E)-beta-farnesene attracted the egg parasitoid Chrysonotomyia ruforum. We tested the behavioural response of C. ruforum females to different concentrations of (E)-beta-farnesene. Egg parasitoids did not respond to this sesquiterpene at either concentration tested. However, they did respond significantly to (E)-beta-farnesene when this compound was offered in combination with the volatile blend emitted from pine twigs without eggs. This response was dependent on the applied concentration of (E) beta-farnesene. Further bioassays with other components [(E)-beta-caryophyllene, delta-cadinene] of the odour blend of pine were conducted in combination with the volatile blend from egg-free pine as background odour. None of the compounds tested against the background of odour from an egg-free pine twig were attractive to the egg parasitoid. These results suggest that the egg parasitoids responded specifically to (E)-beta-farnesene, but only when this compound was experienced in the 'right' context, i.e. when contrasted with a background odour of non oviposition-induced pine volatiles. PMID- 15788713 TI - Learning and discrimination of individual cuticular hydrocarbons by honeybees (Apis mellifera). AB - In social insect colonies, recognition of nestmates, kinship, caste and reproductive status is crucial both for individuals and for the colony. The recognition cues used are thought to be chemical, with the hydrocarbons found on the cuticle of insects often cited as being particularly important. However, in honeybees (Apis mellifera) the role of cuticular hydrocarbons in nestmate recognition is controversial. Here we use the proboscis extension response (PER) conditioning paradigm to determine how well honeybees learn long-chain linear alkanes and (Z)-alkenes present on the cuticle of worker bees, and also how well they can discriminate between them. We found large differences both in learning and discrimination abilities with the different cuticular hydrocarbons. Thus, the tested hydrocarbons could be classified into those which the bees learnt and discriminated well (mostly alkenes) and those which they did not (alkanes and some alkenes). These well-learnt alkenes may constitute important compounds used as cues in the social recognition processes. PMID- 15788715 TI - Attention and memory functioning among pediatric patients with medulloblastoma. AB - OBJECTIVE: To test the hypotheses that memory and attention deficits are prevalent in survivors of childhood medulloblastoma (MB) and that these deficits are associated with problems with academic achievement. METHODS: The medical charts of 38 child survivors of MB, who were administered the California Verbal Learning Test, Child Version (CVLT-C), Conners' Continuous Performance Test (CPT), and the Wechsler Individual Achievement Test (WIAT) as part of a comprehensive neurocognitive test battery, were retrospectively reviewed. RESULTS: Although no significant verbal memory deficits were found, 8 of 11 CPT variables were significantly below the standardization mean (p < or = .01). Additionally, stepwise regression analyses found that increased omission errors were significantly associated with lower reading and math performance (p < or = .01). CONCLUSIONS: These findings confirm previous reports of attention deficits among survivors of MB and provide a better understanding of how the dysfunction of particular attentional substrates (e.g., perceptual sensitivity, response bias) may result in learning problems in this population. PMID- 15788714 TI - The association between maternal cocaine use during pregnancy and physiological regulation in 4- to 8-week-old infants: an examination of possible mediators and moderators. AB - OBJECTIVE: To examine the association between maternal cocaine use during pregnancy and physiological measures of regulation, which included heart rate (HR) and respiratory sinus arrhythmia (RSA). METHODS: Potential mediators and moderators of this association were explored. Participants were 141 mother-infant dyads (77 cocaine exposed and 64 nonexposed) recruited at birth. Average infant HR and RSA was assessed at 4-8 weeks of age during a 15 minute period of sleep. RESULTS: Results indicated a dose-dependent effect of prenatal exposure to cocaine on RSA. There was no evidence that fetal growth or other prenatal exposure to substances mediated this association or that fetal growth or maternal age moderated this association. Regression analyses also indicated that birth weight (BW), but not birthlength (BL), head circumference (HC) or other substance use, mediated the association between prenatal exposure to cocaine and heart rate. CONCLUSIONS: These results suggest that cocaine exposure is associated with physiological regulation at 4-8 weeks of age and highlight the importance of considering level of exposure when assessing infant outcomes. PMID- 15788716 TI - Posttraumatic stress and related impairment in survivors of childhood cancer in early adulthood compared to healthy peers. AB - OBJECTIVE: To compare rates of posttraumatic stress disorder (PTSD) and related impairment between childhood cancer survivors in early adulthood and healthy peers. METHODS: Cancer survivors (n = 57) and comparison group (n = 83) completed measures of PTSD, depression, health-related quality of life (HRQOL), mood, and satisfaction with life (SWL). RESULTS: The cancer survivor group was more likely to have PTSD than the control group (odds ratio = 4.67, p < .05) but was not more likely to experience subclinical PTSD symptoms. The groups differed on physical HRQOL, F(1, 140) = 15.02, p < .001, and positive affect, F(1, 140) = 7.03, p < .01, but did not differ on depression, SWL, psychosocial HRQOL, and negative mood. Those in the survivor group with PTSD (n = 10) experienced more depression and negative affect, worse HRQOL and SWL, perceived their cancer to impact developmental tasks more, and were older at the time of diagnosis compared with those without PTSD (n = 47). CONCLUSIONS: Although most are well adjusted, childhood cancer survivors in early adulthood are more likely to have PTSD and to experience significant impairment compared with healthy peers. PMID- 15788717 TI - Posttraumatic stress symptoms in parents of children with acute burns. AB - OBJECTIVE: To develop a model of risk factors for posttraumatic stress disorder (PTSD) symptoms in parents of children with burns. METHODS: Immediately following the burn and 3 months later, parents reported on their children's and their own psychological functioning and traumatic stress responses. RESULTS: Approximately 47% of the parents reported experiencing significant posttraumatic stress symptoms 3 months after the burn. Our model indicates three independent pathways to PTSD symptoms (i.e., parent-child conflict, parents' dissociation, and children's PTSD symptoms). Additionally, parents' anxiety predicted increased parent-child conflict, conflict with extended family and size of the burn predicted parents' dissociation, and size of the burn and children's dissociation predicted children's PTSD symptoms. CONCLUSIONS: This study suggests that many parents of children with burns suffer from posttraumatic stress symptoms. Interventions that target factors such as family conflict, children's symptoms, and parents' acute anxiety and dissociation may diminish the risk for PTSD. PMID- 15788718 TI - Changes in androgen-mediated reproductive development in male rat offspring following exposure to a single oral dose of flutamide at different gestational ages. AB - Previous studies have indicated that the androgen receptor antagonist, flutamide, can produce a suite of reproductive malformations in the male rat when orally administered daily on gestation days (GD) 12-21. The objective of this study was to investigate the gestation time dependence for the induction of these malformations to establish a robust animal model for future studies of gene expression related to specific malformations. Groups of timed-pregnant Sprague Dawley rats (GD 0 = day of mating) were administered flutamide as a single gavage dose (50 mg/kg) on GD 16, 17, 18, or 19 with 10 dams per group. Control animals (5 dams per time per group) were administered corn oil vehicle (2 ml/kg). Dams were allowed to litter, and their adult male offspring were killed at postnatal day (PND) 100 +/- 10. Anogenital distance was measured at PND 1 and 100. Areolae were scored at PND 13, and permanent nipples evaluated at PND 100. No reproductive tract malformations were found in control male offspring. In the treated groups, malformations were noted following exposure at every GD, although the incidence of specific malformations varied by GD. At GD 16, the highest incidence was noted for permanent nipples (46% pups, 60% litters), epispadias (12% pups, 30% litters), and missing epididymal components (5% pups, 20% litters). The highest incidences for hypospadias (58% pups, 80% litters), vaginal pouch (49% pups, 70% litters), cleft prepuce (29% pups, 60% litters), and missing prostate lobes (12% pups, 60% litters) were noted at GD 17. At GD 18 the highest incidence of malformations noted were epispadias (5% pups, 30% litters), reduced prostate size (32% pups, 90% litters), and abnormal kidneys (3% pups, 30% litters) and bladders (7% pups, 30% litters), while on GD 19 70% of the litters had animals with abnormal seminal vesicles. Testicular and epididymal morphological changes were noted at all GDs and were consistent with the gross observations and peaked in incidence and severity on GD17. The major discrepancy between this study and previous multiple-dose studies was in the very few numbers of animals presenting with cryptorchidism (only one each on GDs 16 and 17), suggesting that exposure over multiple days may be required to induce this malformation. Thus, a single gestational exposure of flutamide induced numerous reproductive tract malformations consistent with previously reports following multiple exposures, with the timing of the exposure producing marked tissue selectivity in the response noted in adult offspring. PMID- 15788719 TI - The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase works as an arsenate reductase in human red blood cells and rat liver cytosol. AB - The mammalian enzymes responsible for reduction of the environmentally prevalent arsenate (AsV) to the much more toxic arsenite (AsIII) are unknown. In the previous paper (Nemeti and Gregus, 2005), we proposed that glyceraldehyde-3 phosphate dehydrogenase (GAPDH) and/or phosphoglycerate kinase (PGK) may catalyze reduction of AsV in human red blood cells (RBC), hemolysate, or rat liver cytosol. In testing this hypothesis, we show here that, if supplied with glutathione (GSH), NAD, and glycolytic substrate, the mixture of purified GAPDH and PGK indeed catalyzes the reduction of AsV. Further analysis revealed that GAPDH is endowed with AsV reductase activity, whereas PGK serves as an auxiliary enzyme, when 3-phosphoglycerate is the glycolytic substrate. The GAPDH-catalyzed AsV reduction required GSH, NAD, and glyceraldehyde-3-phosphate. ADP and ATP moderately, whereas NADH strongly inhibited the AsV reductase activity of the enzyme even in the presence of NAD. Koningic acid (KA), a specific and irreversible inhibitor of GAPDH, inhibited both the classical enzymatic and the AsV-reducing activities of the enzyme in a concentration-dependent fashion. To assess the contribution of GAPDH to the reduction of AsV carried out by hemolysate, rat liver cytosol, or intact erythrocytes, we determined the concentration-dependent effect of KA on AsV reduction by these cells and extracts. Inactivation of GAPDH by KA abolished AsV reduction in intact RBC as well as in the hemolysate and the liver cytosol, when GAPDH in the latter extracts was abundantly supplied with exogenous NAD and glycolytic substrate. However, despite complete inactivation of GAPDH by KA, the hepatic cytosol exhibited significant residual AsV-reducing activity in the absence of exogenous NAD and glycolytic substrate, suggesting that besides GAPDH, other cytosolic enzyme(s) may contribute to AsV reduction in the liver. In conclusion, the key glycolytic enzyme GAPDH can fortuitously catalyze the reduction of AsV to AsIII, if GSH, NAD, and glycolytic substrate are available. AsV reduction may take place during, or as a consequence of, the arsenolytic cleavage of the thioester bond formed between the enzyme's Cys149 and the 3-phosphoglyceroyl moiety of the substrate. Although GAPDH is exclusively responsible for reduction of AsV in human erythrocytes, its role in AsV reduction in vivo remains to be determined. PMID- 15788720 TI - Reduction of arsenate to arsenite by human erythrocyte lysate and rat liver cytosol - characterization of a glutathione- and NAD-dependent arsenate reduction linked to glycolysis. AB - Reduction of arsenate (AsV) to the more toxic arsenite (AsIII) is of high toxicological importance, yet in vivo relevant enzymes involved have not been identified. Purine nucleoside phosphorylase (PNP) is an efficient AsV reductase in vitro, but its role in AsV reduction is irrelevant in vivo. Intact human red blood cells (RBC) possess an AsV reductase activity that is PNP-independent, diminished by depletion of glutathione (GSH), enhanced by oxidants of erythrocytic NAD(P)H, and possibly linked to the lower part of the glycolytic pathway. In order to characterize this PNP-independent AsV reductase activity further, we examined the effects of GSH, inorganic phosphate, some inhibitors of glucose metabolism, glycolytic substrates, and pyridine, as well as adenine nucleotides on AsV reduction in lysed RBC and rat liver cytosol in the presence of BCX-1777, a PNP inhibitor. In hemolysate, GSH enhanced AsV reduction in a concentration-dependent manner, whereas phosphate inhibited it. Glycolytic substrates, especially fructose-1,6-bisphosphate and phosphoglyceric acids, improved AsV reductase activity. NAD, especially together with these substrates, strongly increased AsIII formation, whereas NADH strongly inhibited it. NADP and adenine nucleotides diminished, while 2-phosphoglycollate, which increases the breakdown of the RBC-specific compound 2,3-bisphosphoglycerate to 3 phosphoglycerate, doubled the AsV reductase activity. Although AsV reduction by the liver cytosol responded similarly to GSH, NAD, and glycolytic substrates as in the hemolysate, it was barely influenced by NADH, was diminished by 2 phosphoglycollate, and was stimulated by NADP. Collectively, hemolysate and rat liver cytosol possess a PNP-independent AsV reductase activity. This enzymatic activity requires GSH, NAD, and glycolytic substrates, and purportedly involves one or both of the two functionally linked glycolytic enzymes, glyceraldehyde-3 phosphate dehydrogenase and phosphoglycerate kinase. In addition, the data presented here suggest that yet another PNP-independent AsV reductase resides in the hepatic cytosol. Although this latter enzyme remains unknown, identification of the AsV reductase depending on GSH, NAD, and glycolytic substrates is presented in the following paper. PMID- 15788721 TI - Chronic developmental lead exposure reduces neurogenesis in adult rat hippocampus but does not impair spatial learning. AB - The dentate granule cell (DG) layer of the hippocampal formation has the distinctive property of ongoing neurogenesis that continues throughout adult life. Although the function of these newly generated neurons and the mechanisms that control their birth are unknown, age, activity, diet and psychosocial stress have all been demonstrated to regulate this type of neurogenesis. Little information on the impact of environmental insults on this process has appeared to date. Developmental lead (Pb) exposure has been well documented to impair cognitive function in children and animals and reduce activity-dependent synaptic plasticity in the hippocampus of rodents. Therefore, we examined the effects of this classic environmental neurotoxicant on hippocampal-dependent learning and adult neurogenesis in the hippocampus. Pregnant rats were exposed to a low level of Pb-acetate (0.2%) via the drinking water from late gestation (GD 16) until weaning on postnatal day 21 (PN 21). At weaning, half of the Pb-exposed animals were weaned to control drinking water and the remainder were maintained on Pb water until termination of the study. Animals were paired- housed and on PN 75 were administered a series of injections of a thymidine analog bromodeoxyuridine (BrdU), a marker of DNA synthesis that labels proliferating cells and their progeny. At 12-h intervals for 12 days, rats received an ip injection of BrdU (50 mg/kg). Subjects were sacrificed and perfused 24 h and 28 days after the last injection. Spatial learning was assessed in an independent group of animals beginning on PN 110 using a Morris water maze. No Pb-induced impairments were evident in water maze learning. Immunohistochemistry for the detection of BrdU labeled cells was performed on 40-microm coronal sections throughout the hippocampus. Continuous exposure to Pb (Life) reduced the total number of BrdU positive cells at 28 days without affecting the total number of labeled cells evident 24 h after the last injection. No differences in the number of progenitor cells labeled or surviving were seen between control and treated animals whose Pb exposure was terminated at weaning. Double labeling with BrdU and the glial specific marker, glial acidic fibrillary protein (GFAP) indicated that the bulk of the surviving cells were of a neuronal rather than a glial phenotype. These data reveal that chronic low-level Pb exposure reduces the capacity for neurogenesis in the adult hippocampus. Despite deficits in synaptic plasticity previously reported from our laboratory, and now structural plasticity, no significant impact on spatial learning was detected. PMID- 15788722 TI - Post-transcriptional regulation of metallothionein isoform 1 and 2 expression in the human breast and the MCF-10A cell line. AB - Studies have shown, using immunohistochemical staining, that the MT-1 and MT-2 proteins (MT-1/2) are overexpressed in a substantial subset of ductal breast cancers, that overexpression occurs early in the disease process, and that this overexpression is indicative of a poor prognosis. Normal ductal breast epithelium fails to immunostain for the MT-1/2 protein, whereas the myoepithelial cells of the ducts stain intensely. There is no information regarding the expression of the mRNAs for the eight active MT-1 and MT-2 genes in normal breast duct epithelium. Microdissection of normal breast samples was used to obtain total RNA from enriched populations of ductal epithelium and myoepithelium. Analysis by reverse-transcription polymerase chain reaction (RT-PCR) demonstrated that the identity of the MT isoform-specific genes expressed (MT-2A and MT-1X) and their relative levels of expression were similar between the myoepithelial and ductal components. These findings indicate that the ductal and myoepithelial components express similar amounts of MT-2A and MT-1X mRNAs, but that they have distinctly different expression of the MT-1/2 protein. Confluent cultures of MCF-10A breast epithelial cells were exposed to Cd(+2) to test for evidence of post transcriptional regulation of MT-1/2 protein accumulation in ductal epithelium. It was demonstrated that Cd(+2) elicited only a marginal induction of MT-1E, MT 1X, or MT-2A mRNAs, whereas, there was a marked increase in MT-1/2 protein, reaching levels of 6% of total cell protein under conditions of extended exposure. This study suggests that the mechanism underlying the finding of increased MT-1/2 protein expression in ductal breast cancer may involve, to some degree, the post-transcriptional regulation of MT-1/2 protein expression. PMID- 15788723 TI - Identification of the tryptophan photoproduct 6-formylindolo[3,2-b]carbazole, in cell culture medium, as a factor that controls the background aryl hydrocarbon receptor activity. AB - The presence of high affinity ligands for the aryl hydrocarbon receptor (AhR) in cell culture medium has generally been overlooked. Such compounds may confound mechanistic studies of the important AhR regulatory network. Numerous reports have described that light exposed cell culture medium induces AhR-dependent activity. In this study, we aimed at identifying the causative substance(s). A three-dimensional factorial design was used to study how the background activity of CYP1A1 in a rat hepatoma cell line (MH1C1) was controlled by photoproducts formed in the medium exposed to normal laboratory light. The light induced activity was found to be tryptophan dependent, but independent of riboflavin and other components in the medium. The light exposed medium showed the same transient enzyme inducing activity in vitro as the AhR ligand 6-formylindolo[3,2 b]carbazole (FICZ). This substance, which we have previously identified as being formed in UV-exposed tryptophan solutions, is a substrate for CYP1A1 and it has a higher AhR binding affinity than TCDD. Several tryptophan related photoproducts were detected in the light-exposed medium. For the first time one of the formed photoproducts was identified as FICZ with bioassay driven fractionation coupled with HPLC/MS. These results clearly show that tryptophan derived AhR ligands, which have been suggested to be endogenous AhR ligands, influence the background levels of CYP1A1 activity in cells in culture. PMID- 15788724 TI - The effects of early lead exposure on the brains of adult rhesus monkeys: a volumetric MRI study. AB - Little is known about direct effects of exposure to lead on central nervous system development. We conducted volumetric MRI studies in three groups of 17 year-old rhesus monkeys: (1) a group exposed to lead throughout gestation (n = 3), (2) a group exposed to lead through breast milk from birth to weaning (n = 4), and (3) a group not exposed to lead (n = 8). All fifteen monkeys were treated essentially identically since birth with the exception of lead exposure. The three-dimensional MRI images were segmented on a computer workstation using pre tested manual and semi-automated algorithms to generate brain volumes for white matter, gray matter, cerebrospinal fluid, and component brain structures. The three groups differed significantly in the adjusted (for total brain size) volumes of the right cerebral white matter and the lateral ventricles. A significant reduction was noted in right cerebral white matter in prenatally exposed monkeys as compared to controls (p = 0.045). A similar reduction was detected in the white matter of the contralateral hemisphere; however, this difference did not achieve statistical significance (p = 0.143). Prenatally exposed monkeys also had larger right (p = 0.027) and left (p = 0.040) lateral ventricles. Depending on the timing of exposure during development, lead may exhibit differential effects with resultant life-long alterations in brain architecture. PMID- 15788726 TI - Miotic tolerance to sarin vapor exposure: role of the sympathetic and parasympathetic nervous systems. AB - O-isopropyl methylphosphonofluoridate, also known as sarin or GB, is a highly toxic organophosphorous compound that exerts its effect by inhibiting the enzyme acetylcholinesterase. While the effects of a single exposure to GB vapor are well characterized, the effects of multiple exposures to GB vapor are less clear. Previous studies in the rat and guinea pig have demonstrated that multiple exposures result in tolerance to the miotic effect of nerve agents. The aim of the present study was to examine potential mechanisms responsible for tolerance to the miotic effect of GB vapor that has been observed in the rat after multiple exposures. Multiple whole-body inhalation exposures to GB vapor were conducted in a dynamic airflow chamber. Exposures lasted 60 min and each of the three exposures occurred at 24-h intervals. The results of the present study demonstrate that the alpha-adrenergic antagonist phentolamine and the beta adrenergic receptor antagonist propranolol did not affect the development of tolerance to the miotic effect of GB vapor, suggesting that enhanced sympathetic tone to the eye is not responsible for the observed tolerance. Administration of atropine before the first exposure prevented the tolerance to the miotic effect of GB vapor after the third exposure, suggesting that the tolerance is the result of muscarinic receptor desensitization secondary to receptor stimulation. The present study extends the findings of previous studies to strengthen the hypothesis that the miotic tolerance observed in the rat upon repeated exposure to nerve agents is due to desensitization of muscarinic acetylcholine receptors located on the pupillary sphincter. PMID- 15788725 TI - Dose thresholds should exist for chemical carcinogens. PMID- 15788727 TI - Perinatal exposure to the fungicide prochloraz feminizes the male rat offspring. AB - Prochloraz is a commonly used fungicide that has shown multiple mechanisms of action in vitro. It antagonizes the androgen and the estrogen receptors, agonizes the Ah receptor, and inhibits aromatase activity. In vivo prochloraz acts antiandrogenically in the Hershberger assay by reducing weights of reproductive organs, affecting androgen-regulated gene expressions, and increasing luteinizing hormone (LH) levels. The purpose of this study was to investigate reproductive toxic effects after exposure during gestation and lactation to prochloraz alone and a mixture of five pesticides (deltamethrin, methiocarb, prochloraz, simazine, and tribenuron-methyl). Prochloraz (30 mg/kg/day) or the mixture (20 mg/kg/day) was dosed to pregnant Wistar dams from gestational day (GD) 7 until postnatal day (PND) 16. Some dams were taken for cesarean section at GD 21, and others were allowed to give birth. Results showed that prochloraz and the mixture significantly reduced plasma and testicular testosterone levels in GD 21 male fetuses, whereas testicular progesterone was increased. Gestational length was increased by prochloraz. Chemical analysis of the rat breast milk showed that prochloraz was transferred to the milk. In males a significant increase of nipple retention was found, and the bulbourethral gland weight was decreased, whereas other reproductive organs were unaffected. In addition cytochrome P450 (CYP)1A activities in livers were induced by prochloraz, possibly as a result of Ah receptor activation. Behavioral studies showed that the activity level and sweet preference of adult males were significantly increased. Overall these results strongly indicate that prochloraz feminizes the male offspring after perinatal exposure, and that these effects are due, at least in part, to diminished fetal steroidogenesis. PMID- 15788728 TI - DNA interaction and dual topoisomerase I and II inhibition properties of the anti tumor drug prodigiosin. AB - Prodigiosin is a red pigment produced by Serratia marcescens with apoptotic activity. We examined the mechanism of action of this tripyrrole alkaloid, focusing on its interaction with DNA and its ability to inhibit both topoisomerase I and topoisomerase II. We also evaluated the DNA damage induced in cancer cell lines. Prodigiosin-DNA intercalation was analyzed using a competition dialysis assay with different DNA base sequences. Topoisomerase I and II inhibition was studied in vitro by a cleavage assay, and in cultured cells, by analysis of its ability to form covalent complexes. Furthermore, we analyzed DNA damage by pulse-field gel electrophoresis and by immunocytochemistry. Apoptosis inducing factor (AIF)/phospho-H2AX (p-H2AX) double labeling by confocal microscopy was performed to determine the possible implication of AIF in the prodigiosin-DNA damage. Finally, we studied the ability of this drug to induce copper-mediated DNA damage at different pH by a DNA cleavage assay. Our results demonstrate prodigiosin-DNA interaction in vitro and in cultured cells. It involves prodigiosin-DNA intercalation, with some preference for the alternating base pairs but with no discrimination between AT or CG sequences, dual abolition of topoisomerase I and II activity and, as consequence, DNA cleavage. Prodigiosin DNA damage is independent of AIF. Furthermore, we found that copper-mediated cleavage activity is associated with pH (occurring at pH 6.8 rather than pH 7.4) and with the Cu(2+) ion concentration. These results indicate DNA a therapeutic target for prodigiosin and could explain the apoptosis mechanism of action induced by this antineoplastic drug. PMID- 15788729 TI - Analysis of somatic molecular changes, clinicopathological features, family history, and germline mutations in colorectal cancer families: evidence for efficient diagnosis of HNPCC and for the existence of distinct groups of non HNPCC families. AB - OBJECTIVE: To analyse somatic molecular changes, clinicopathological features, family history, and germline mutations in families with colorectal cancer (CRC). METHODS: Molecular changes (K-ras and beta-catenin mutations, chromosome 18q allele loss (LOH), APC LOH, microsatellite instability (MSI), and expression of beta-catenin and p53) were examined in four series of CRC patients with proven or probable hereditary disease: hereditary non-polyposis colon cancer (HNPCC); MYH associated polyposis (MAP); multiple (>5) colorectal adenomas without familial adenomatous polyposis (FAP); and other families/cases referred to family cancer clinics (FCC series). HNPCC was diagnosed using a combination of germline mutation screening and tumour studies. A series of unselected CRC patients was also studied. RESULTS: There was overlap between genetic pathways followed by each type of CRC, but significant differences included: increased frequency of K ras mutation and reduced frequency of APC LOH in cancers from MAP, but not from multiple adenoma patients; reduced frequency of LOH in HNPCC CRCs; and increased MSI in CRCs from HNPCC, but not from FCC or multiple adenoma patients. HNPCC was apparently detected efficiently by combined germline and somatic analysis. Cancers from the FCC, unselected, and multiple adenoma series shared similar molecular characteristics. In the FCC and multiple adenoma series, hierarchical cluster analysis using the molecular features of the cancers consistently identified two distinct groups, distinguished by presence or absence of K-ras mutation. CONCLUSIONS: While K-ras mutation status is known to differentiate hereditary bowel cancer syndromes such as MAP and FAP, it may also distinguish groups of non-HNPCC, FCC patients whose disease has different, as yet unknown, genetic origins. PMID- 15788731 TI - Putting our research into prospective. PMID- 15788732 TI - Evolgate fixation of doubled flexor graft in anterior cruciate ligament reconstruction: biomechanical evaluation with cyclic loading. AB - BACKGROUND: The weakest points in hamstrings anterior cruciate ligament reconstruction are its points of fixation, especially on the tibial side. Methods for graft fixation to bone should be strong enough to avoid failure, stiff enough to restore load-displacement response, and secure enough to resist slippage under cyclic loading. HYPOTHESIS: Biomechanical properties of the interference screw can be improved by reinforcing the walls of the tunnel with a metal spiral (Evolgate fixation). STUDY DESIGN: Controlled laboratory study. METHODS: Three paired tests were performed using common digital extensor bovine tendons fixed to porcine tibias with interference screw, Intrafix, and Evolgate; critical graft fixation conditions were simulated by applying subsequent cycles to the graft before loading the graft until failure. RESULTS: The strength, stiffness, and resistance to slippage of the Evolgate were significantly higher than those of the interference screw; the ultimate failure load of the Evolgate was significantly higher than that of the Intrafix (1058+/-130 N and 832+/-156 N, respectively; P=.02). No statistically significant differences were found between Evolgate and Intrafix regarding the stiffness and slippage under cyclic load. CONCLUSION: Evolgate fixation seems to be stronger, stiffer, and more resistant to slippage than is interference screw fixation of similar length, and it is stronger but not stiffer than Intrafix. CLINICAL RELEVANCE: The Evolgate provides structural properties that seem to be appropriate for an intensive rehabilitation after anterior cruciate ligament reconstruction using a doubled semitendinosus and gracilis graft. PMID- 15788730 TI - RAI1 variations in Smith-Magenis syndrome patients without 17p11.2 deletions. AB - BACKGROUND: Smith-Magenis syndrome (SMS) (OMIM No 182290) is a mental retardation syndrome characterised by behavioural abnormalities, including self injurious behaviours, sleep disturbance, and distinct craniofacial and skeletal anomalies. It is usually associated with deletion involving 17p11.2 and is estimated to occur in 1/25,000 births. Heterozygous frameshift mutations leading to protein truncation in retinoic acid induced 1 gene (RAI1) have been identified in individuals with phenotypic features consistent with SMS. RAI1 lies within the 17p11.2 locus, but these patients did not have 17p11.2 deletions. OBJECTIVE: Analysis of four individuals with features consistent with SMS for variations in RAI1, using a polymerase chain reaction and sequencing strategy. None of these patients carry 17p11.2 deletions. RESULTS: Two patients had small deletions in RAI1 resulting in frameshift and premature truncation of the protein. Missense mutations were identified in the other two. Orthologs across other genomes showed that these missense mutations occurred in identically conserved regions of the gene. The mutations were de novo, as all parental samples were normal. Several polymorphisms were also observed, including new and reported SNPs. The patients' clinical features differed from those found in 17p11.2 deletion by general absence of short stature and lack of visceral anomalies. All four patients had developmental delay, reduced motor and cognitive skills, craniofacial and behavioural anomalies, and sleep disturbance. Seizures, not previously thought to be associated with RAI1 mutations, were observed in one patient of the cohort. CONCLUSIONS: Haploinsufficiency of the RAI1 gene is associated with most features of SMS, including craniofacial, behavioural, and neurological signs and symptoms. PMID- 15788733 TI - The use of knee braces, part 1: Prophylactic knee braces in contact sports. AB - Surrogate knee model biomechanical studies have indicated that off-the-shelf braces provide 20% to 30% greater resistance to a lateral blow when the knee is in full extension. Custom functional braces doubled the protective effects and proved effective with the knee in some flexion. Although functional performance studies are not consistent, preventive knee braces may slow straight-ahead sprint speed, cause early fatigue, and increase muscular relaxation pressures, energy expenditure, blood lactate levels, maximal torque output, oxygen consumption, and heart rate. Two epidemiologic studies have been performed. At West Point, a randomized control study of 71 injuries in 1396 cadets indicated knee brace effectiveness with a statistically higher rate of injury in the control group (3.4/1000 exposures) than in the braced group (1.5/1000 exposures), with the most significance for medial collateral ligament sprains in defensive players. The Big Ten Conference conducted a descriptive study of 100 medial collateral ligament sprains among 987 players in different positions, strings, and types of session. Brace-wear tendency varied directly with the unbraced player counterpart's risk of medial collateral ligament sprain, with the nonplayer linemen experiencing both the greatest risk of unbraced practice session injury (0.0801 injuries/1000 exposures) and the highest incidence of brace wear (85%). During practices, there was a nonsignificant but very consistent reduction in injury rate for braced players in every position and string. During games, there was also a reduced rate for linemen and the linebacker/tight end group. The study concluded that although the issue is not closed, preventive knee braces appear to offer some protection to the medial collateral ligament from a contact injury involving a valgus blow, but there may be negative effects on performance level, leg cramping, and fatigue symptoms. PMID- 15788734 TI - The genetics of cardiac arrhythmias. AB - Cardiac rhythm problems result in high levels of morbidity and mortality, with sudden arrhythmic death claiming approximately 300,000 lives in the United States each year. Investigations into the genetic contributions to rhythm and conduction disorders have found genes or loci associated with primary rhythm/conduction disorders such as familial atrial fibrillation and atrio-ventricular block, underscoring the importance of collecting a thorough family history. Combinations of single or multiple genes and environmental risk factors may place only certain family members at risk. Some cardiac muscle problems, such as cardiomyopathy, predispose to arrhythmia and have documented genetic components. Primary health care providers need current knowledge of genetic contributions to rhythm/conduction problems so that family members at risk can be identified early and cared for appropriately. This article provides an overview of the genetic contributions to cardiac rhythm and conduction problems. PMID- 15788735 TI - Introducing the MUC16 gene: implications for prevention and early detection in epithelial ovarian cancer. AB - More than 24,000 women in the United States are diagnosed with ovarian cancer every year, and half of these women die from their disease. Stage 1 ovarian cancer is curable in 95% of cases; however, due to inadequate screening tools and lack of symptoms in early disease, ovarian cancer is generally at Stage 3 or 4 when finally diagnosed. CA125 is a tumor antigen used to monitor the progression and regression of epithelial ovarian cancer. When its levels are elevated postsurgery (hysterectomy/salpingo-oophorectomy with or without peritoneal washings and lymph node biopsy) and postchemotherapy, it is suggestive of recurrent disease. Due to its similarly elevated levels in some nonmalignant conditions, however, it is not specific enough to be used for population screening. The CA125 molecule is considered a very large glycoprotein because of its molecular weight, and it has three domains: the carboxy terminal domain, the extracellular domain, and the amino terminal domain. MUC16 is the gene that encodes the peptide moiety of the CA125 molecule. MUC16 domains provide novel opportunities to develop new assays and refine current tools to improve the sensitivity and specificity of CA125 for population-based screening guidelines. PMID- 15788736 TI - A comparison of gastric and rectal CO2 in cardiac surgery patients. AB - Critical care nurses assess and treat clinical conditions associated with inadequate oxygenation. Changes in regional organ (gut) blood flow are believed to occur in response to a decrease in oxygenation. Although the stomach is a widely accepted monitoring site, there are multiple methodological and measurement issues associated with the gastric environment that limit the accuracy of P CO2 detection. The rectum may provide nurses with an alternative site for monitoring changes in P CO2 without the limitations associated with gastric monitoring. This pilot study used a repeated measures design to examine changes in gastric and rectal P CO2 during elective coronary artery bypass grafting with cardiopulmonary bypass (CPB) and in the immediate 4-hr postoperative period in 26 subjects. The systemic indicators explained little variation in the regional indicators during protocol. A comparison of rectal and gastric P CO2 revealed no statistically significant differences in the direction or magnitude of change over any phase of cardiac surgery (baseline, CPB, post CPB). A reduction in both rectal and gastric P CO2 occurred during CPB, and both values trended upward during the post-CPB phase. However, poor correlation and agreement was found between the measures of P CO2 at the two sites. Although clinically important, the cause is unclear. Possible explanations include variation in CO2 production between the gastric and rectal site, differences in sensitivity of the two monitoring instruments, or the absence of hemodynamic complications, which limited the extent of change in P CO2. Further investigation using patients with more profound changes in oxygenation are needed to identify response patterns and possible mechanisms. PMID- 15788737 TI - Leukocyte-platelet aggregates in rat peripheral blood after ischemic stroke and reperfusion. AB - Ischemic stroke and reperfusion (ISR) is associated with an inflammatory response characterized, in part, by the formation of leukocyte-platelet aggregates (LPA). Aggregate formation may amplify the immunologic and hemostatic functions of both cell types and thus exacerbate reperfusion injury after ischemic stroke. LPA formation in peripheral blood may also serve as a biomarker of the severity of injury. However, it is not fully known whether ISR causes LPA formation that can be detected in the peripheral blood. Therefore, the purpose of this study was to measure LPA in the peripheral blood after ISR using a rat model. The filament method was used to perform ISR. Blood was collected from the jugular vein before ischemia, after 4 hours of ischemia, and after 1 hour of reperfusion. Flow cytometry was used to quantify LPA in peripheral blood. Separate ISR groups were treated with tirofiban, a platelet GPIIb/IIIa inhibitor, and fucoidan, a selectin adhesion molecule inhibitor, and analyzed for LPA. Leukocyte CD11b expression and reactive oxygen species production were also analyzed to note the role of polymorphonuclear neutrophilic (PMN) activation on LPA formation. After ISR, LPA levels in peripheral blood were twice as large as preischemic levels. Both GPIIb/IIIa and selectin adhesion molecule inhibition (p < .05) decreased LPA to preischemic values. PMN CD11b expression was increased above baseline but did not differ between groups. Reactive oxygen species production did not differ between groups during reperfusion. These data suggest that ischemic stroke and reperfusion results in an increase in LPA that can be consistently measured in peripheral blood. LPA formation may be a useful biomarker and potential therapeutic target after ischemic stroke and reperfusion. PMID- 15788738 TI - Arginine supplementation does not enhance serum nitric oxide levels in elderly nursing home residents with pressure ulcers. AB - The purpose of this study was to determine whether arginine supplementation enhances in vitro (neutrophil burst and mitogen-induced lymphocyte proliferation) and in vivo (delayed-type hypersensitivity [DTH] and serum nitric oxide) measures of immune function in nursing home elders with pressure ulcers. Twenty-six elders, 65 years of age or older, with one or more pressure ulcers, were randomized to receive 8.5 g of arginine or an isonitrogenous supplement for 4 weeks. Immune function studies and serum arginine, ornithine, citrulline, and nitric oxide were measured at baseline, 4 weeks postsupplementation (Week 4) and after a 6-week washout (Week 10). At Week 4, serum ornithine increased (p = .01) and arginine trended to increase (p = .055), but there was no increase in citrulline or nitric oxide with arginine supplementation. There were no differences in neutrophil burst or DTH responses between groups. Whole blood mitogen-induced proliferation decreased significantly at Week 10 in the isonitrogenous but not in the arginine-supplemented group. There is mounting concern that arginine supplementation during an inflammatory state could be detrimental due to overwhelming nitric oxide production. A key finding of this study is that arginine supplementation did not increase serum nitric oxide levels over that observed in elders with pressure ulcers given an isonitrogenous supplement. PMID- 15788739 TI - Role for up-regulated ganglioside biosynthesis and association of Src family kinases with microdomains in retinoic acid-induced differentiation of F9 embryonal carcinoma cells. AB - Mouse F9 embryonal carcinoma cells have been widely used as a model for studying the mechanism of embryonic differentiation, because they are similar to the inner cell mass of early mouse embryos and can differentiate into primitive endoderm (PrE) following retinoic acid (RA) treatment. During F9 cell differentiation, the carbohydrate chains of glycoproteins and their corresponding glycosyltransferases are known to undergo rapid changes. However, there have been no corresponding reports on the expression of gangliosides. We have developed a custom cDNA array that is highly sensitive for the genes responsible for sphingolipid (SL) biosynthesis and metabolism. Using this, we found that, of the 28 selected genes, 26 exhibited increased expression during F9 differentiation into PrE. Although neutral glycosphingolipids (GSLs) were expressed at similar levels before and after differentiation, a greater than 20-fold increase in total ganglioside content was evident in PrE. Glucosylceramide synthase inhibitors (d-threo-1 phenyl-2-decanoylamino-3-morpholino-1-propanol [d-PDMP] and its analog) depleted gangliosides and this resulted in delayed expression of Disabled-2 (Dab-2), suggesting the involvement of gangliosides in F9 cell differentiation. Disruption of cholesterol-enriched membrane microdomains by methyl-beta-cyclodextrin (MbetaCD) also delayed differentiation. Both MbetaCD and d-PDMP blocked the accumulation of Src family kinases (SFKs) to microdomains. However, d-PDMP did not block flotillin accumulation, yet MbetaCD did. Additionally, confocal laser microscopy revealed the formation of distinct functional microdomains integrating SFKs with gangliosides and cholesterol during PrE differentiation. Thus, we demonstrate the outstanding up-regulation of ganglioside biosynthesis and its importance in the formation of distinct microdomains incorporating SFKs with gangliosides during RA-induced differentiation of F9 cells. PMID- 15788740 TI - High constitutive activity of a virus-encoded seven transmembrane receptor in the absence of the conserved DRY motif (Asp-Arg-Tyr) in transmembrane helix 3. AB - The highly conserved Arg in the so-called DRY motif (Asp-Arg-Tyr) at the intracellular end of transmembrane helix 3 is in general considered as an essential residue for G protein coupling in rhodopsin-like seven transmembrane (7TM) receptors. In the open reading frame 74 (ORF74) receptor encoded by equine herpesvirus 2 (EHV2), the DRY motif is substituted with a DTW motif. Nevertheless, this receptor signaled with high constitutive activity through Gi as determined by a receptor-mediated inhibition of forskolin-induced cAMP production and by an induction of the serum response element-driven transcriptional activity through a pertussis toxin-sensitive manner. Gs and Gq were not activated constitutively as determined by the lack of inositol phosphate turnover and activities of the three transcription factors: cAMP response element binding protein (CREB), nuclear factor-kappaB, and nuclear factor of activated T cells. Coexpression of the ORF74-EHV2 receptor with the promiscuous G protein Gqi4myr supported the constitutive Gi activation as determined by inositol phosphate turnover and CREB activation. The constitutive activity was inhibited by nonpeptide inverse agonists with micromolar potencies, and the chemokine CXCL6 acted as a high-affinity agonist. It is noteworthy that reconstitution of the DRY motif resulted in a 4- to 5-fold decrease of the constitutive activity. Both the wild type and the receptor with the reconstituted DRY motif were expressed at the cell surface as indicated by immunohistochemistry and enzyme-linked immunosorbent assay analysis. It is concluded that the Arg of the DRY motif in transmembrane helix 3 is not essential for G protein coupling based on the constitutive as well as the ligand-mediated activity observed for ORF74-EHV2. PMID- 15788741 TI - Expression, pharmacological profile, and functional coupling of A2B receptors in a recombinant system and in peripheral blood cells using a novel selective antagonist radioligand, [3H]MRE 2029-F20. AB - In this study, we compared the pharmacological and biochemical characteristics of A(2B) adenosine receptors in recombinant (hA(2B)HEK293 cells) and native cells (neutrophils, lymphocytes) by using a new potent 8-pyrazole xanthine derivative, [(3)H]N-benzo[1,3]dioxol-5-yl-2-[5-(1,3-dipropyl-2,6-dioxo-2,3,6,7-tetrahydro-1H purin-8-yl)-1-methyl-1H-pyrazol-3-yl-oxy]-acetamide] ([(3)H]MRE 2029-F20), that has high affinity and selectivity for hA(2B) versus hA(1),hA(2A), and hA(3) subtypes. [(3)H]MRE 2029-F20 bound specifically to the hA(2B) receptor stably transfected in human embryonic kidney (HEK) 293 cells with K(D) of 2.8 +/- 0.2 nM and B(max) of 450 +/- 42 fmol/mg of protein. Saturation experiments of [(3)H]MRE 2029-F20 binding in human neutrophils and lymphocytes detected a single high affinity binding site with K(D) values of 2.4 +/- 0.5 and 2.7 +/- 0.7 nM, respectively, and B(max) values of 79 +/- 10 and 54 +/- 8 fmol/mg of protein, respectively, in agreement with real-time reverse transcription polymerase chain reaction studies showing the presence of A(2B) mRNA. The rank order of potency of typical adenosine ligands with recombinant hA(2B) receptors was consistent with that typically found for interactions with the A(2B) subtype and was also similar in peripheral blood cells. 5'-N-Ethyl-carboxamidoadenosine stimulated cAMP accumulation in both hA(2B)HEK293 and native cells, whereas phospholipase C activation was observed in recombinant receptors and endogenous subtypes expressed in neutrophils but not in lymphocytes. MRE 2029-F20 was revealed to be a potent antagonist in counteracting the agonist effect in both signal transduction pathways. In conclusion, [(3)H]MRE 2029-F20 is a selective and high affinity radioligand for the hA(2B) adenosine subtype and may be used to quantify A(2B) endogenous receptors. In this work, we demonstrated their presence and functional coupling in neutrophils and lymphocytes that play a role in inflammatory processes in which A(2B) receptors may be involved. PMID- 15788743 TI - A first assessment of genetic variation among Morchella esculenta (morel) populations. AB - Habitat loss and fragmentation have serious consequences for species diversity as well as genetic diversity within a species. As the most sought-after culinary fungus in the Midwest United States, morels (Morchella esculenta and related species) demand the attention of conservationists interested in preserving biological and genetic diversity. Little is known about the natural history of M. esculenta, which is critical information for understanding population dynamics as well as the impacts of habitat fragmentation and harvesting. We report initial results from our long-term studies of genetic variability among fruiting bodies at the Conard Environmental Research Area at Grinnell College, Grinnell, Iowa. Using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), a technique that has been successfully used to examine intrapopulation structure and detect clonal populations in numerous fungi, we found substantially higher levels of genetic polymorphism among 57 fruiting bodies than has been previously reported. Though laboratory studies indicate that the inbreeding potential for this fungus is high, we found little evidence for inbreeding, with only two pairs of the randomly chosen isolates having identical genotypes at the 34 loci examined. This work highlights the importance of further attempts to resolve important aspects of the morel life cycle regarding heterokaryosis and inbreeding potential. PMID- 15788742 TI - Differential effects of Gq alpha, G14 alpha, and G15 alpha on vascular smooth muscle cell survival and gene expression profiles. AB - Gqalpha family members (Gqalpha, G11alpha, G14alpha, and G15/16alpha) stimulate phospholipase Cbeta (PLCbeta) and inositol lipid signaling but differ markedly in amino acid sequence and tissue distribution predicting unappreciated functional diversity. To examine functional differences, we compared the signaling properties of Gqalpha, G14alpha, and G15alpha and their cellular responses in vascular smooth muscle cells (VSMC). Constitutively active forms of Gqalpha, G14alpha, or G15alpha elicit markedly different responses when introduced to VSMC. Whereas each Galpha stimulated PLCbeta to similar extents when expressed at equal protein levels, Gqalpha and G14alpha but not G15alpha initiated profound cell death within 48 h. This response was the result of activation of apoptotic pathways, because Gqalpha and G14alpha, but not G15alpha, stimulated caspase-3 activation and did not alter phospho-Akt, a regulator of cell survival pathways. Gqalpha and G14alpha stimulate nuclear factor of activated T cell (NFAT) activation in VSMC, but Galpha-induced cell death seems independent of PKC, InsP(3)/Ca(2+), and NFAT, in that pharmacological inhibitors of these pathways did not block cell death. Gene expression analysis indicates that Gqalpha, G14alpha, and G15alpha each elicit markedly different profiles of altered gene sets in VSMC after 24 h. Whereas all three Galpha stimulated changes (> or =2 fold) in 50 shared mRNA, Gqalpha and G14alpha (but not G15alpha) stimulated changes in 221 shared mRNA, many of which are reported to be pro-apoptotic and/or involved with TNF-alpha signaling. We were surprised to find that each Galpha also stimulated changes in nonoverlapping Galpha-specific gene sets. These findings demonstrate that Gqalpha family members activate both overlapping and distinct signaling pathways and are more functionally diverse than previously thought. PMID- 15788744 TI - Spliced leader RNA-mediated trans-splicing in phylum Rotifera. AB - In kinetoplastids, Euglena, and four metazoan phyla, trans-splicing has been described as a mechanism for the generation of mature messenger RNAs (mRNAs): 5' ends of precursor mRNAs are replaced by a short spliced leader (SL) exon from a small SL RNA. Although the full phylogenetic range is unknown, trans-splicing has not been found in vertebrates, insects, plants, or yeast. In animal groups where it does occur, i.e., nematodes, cnidarians, platyhelminths, and primitive chordates, SL RNAs do not show sequence relatedness across phyla. The apparently sporadic phylogenetic distribution and the lack of SL RNA homology have led to opposing hypotheses on its evolution, involving either an ancient origin followed by loss in multiple lineages or independent acquisition in several taxa. Here we present evidence for the occurrence of trans-splicing in bdelloid rotifers (Bdelloidea, Rotifera). A common 23-nt sequence, representing the SL exon diagnostic of SL RNA-mediated trans-splicing-was found at the 5'-end of at least 50%-65% of mRNAs from Adineta ricciae and Philodina sp. The trans-splicing pattern in bdelloid rotifers can be unusually complex, as observed in transcripts from a heat shock protein gene, hsp82-1, where the SL exon was spliced to three alternative positions. Bdelloid rotifer SL RNAs were found to be 105 or 106 nt long and comprised the SL sequence, a conserved splice donor site and an intron containing a putative spliceosome-binding motif. Intriguingly, some similarity of rotifer SL RNA sequence and predicted secondary structure was seen to that of the predominant SL1 RNA of nematodes, although it is unlikely that this demonstrates homology. In addition, sequence corresponding to the rotifer SL exon was found at the 5'-end of a number of full-length complementary DNA (cDNA) clones in a rice (Oryza sativa) database. None of these cDNAs gave a close match with homologous plant genes, suggesting that a small but significant portion of the rice expressed sequence tag database represents sequences derived from rotifers. In summary, the description of SL-mediated trans-splicing in Rotifera extends its representation to at least five metazoan phyla, making it increasingly probable that this is a phylogenetically widespread and therefore ancient phenomenon. PMID- 15788745 TI - mRNA-mediated intron losses: evidence from extraordinarily large exons. AB - Multicellular eukaryotes that have high intron density have their introns almost evenly distributed within genes, but unicellular eukaryotes that are generally intron poor have their introns asymmetrically distributed toward the 5' ends of genes. This was explained by homologous recombination of genomic DNA with the cDNA reverse transcribed from the 3' polyadenylated tail of spliced mRNA. This paper is to study whether mRNA-mediated intron losses have ever occurred in multicellular eukaryotes. If intron losses were mRNA-mediated, adjacent introns should be commonly lost together. A direct result is fusion of several previously adjacent exons and producing a large exon. We found that extraordinarily large exons (ELEs) are common not only in unicellular eukaryotes but also in multicellular eukaryotes. The percentage of genes having ELEs is negatively correlated with intron abundance. In addition, the number of lost introns estimated from the relative lengths of ELEs is negatively correlated with the number of extant introns. These results support mRNA-mediated intron losses in all eukaryotes. Moreover, we found that the ELEs of intron-common eukaryotes (with more than 0.5 intron per gene on average) are not only located at 3' ends but also at 5' ends and the middle of genes. This is contrary to what would be expected if the involved cDNAs were reverse transcribed from the 3' polyadenosine ends. A remarkable difference in intron distribution was revealed between intron rare eukaryotes and intron-common eukaryotes. The intron-rare eukaryotes show very strong 5'-biased intron distribution, whereas the intron-common eukaryotes display even intron distribution or only weak 5'-biased distribution. We suspected that intron losses from 3' end of genes may be limited in intron-rare eukaryotes. The intron losses from intron-common eukaryotes should have other priming mechanism, like self-primed reverse transcription. PMID- 15788746 TI - Conservation versus parallel gains in intron evolution. AB - Orthologous genes from distant eukaryotic species, e.g. animals and plants, share up to 25-30% intron positions. However, the relative contributions of evolutionary conservation and parallel gain of new introns into this pattern remain unknown. Here, the extent of independent insertion of introns in the same sites (parallel gain) in orthologous genes from phylogenetically distant eukaryotes is assessed within the framework of the protosplice site model. It is shown that protosplice sites are no more conserved during evolution of eukaryotic gene sequences than random sites. Simulation of intron insertion into protosplice sites with the observed protosplice site frequencies and intron densities shows that parallel gain can account but for a small fraction (5-10%) of shared intron positions in distantly related species. Thus, the presence of numerous introns in the same positions in orthologous genes from distant eukaryotes, such as animals, fungi and plants, appears to reflect mostly bona fide evolutionary conservation. PMID- 15788747 TI - Solution structure of a modified 2',5'-linked RNA hairpin involved in an equilibrium with duplex. AB - The isomerization of phosphodiester functionality of nucleic acids from 3',5'- to a less common 2',5'-linkage influences the complex interplay of stereoelectronic effects that drive pseudorotational equilibrium of sugar rings and thus affect the conformational propensities for compact or more extended structures. The present study highlights the subtle balance of non-covalent forces at play in structural equilibrium of 2',5'-linked RNA analogue, 3'-O-(2-methoxyethyl) substituted dodecamer *CG*CGAA*U*U*CG*CG, 3'-MOE-2',5'-RNA, where all cytosines and uracils are methylated at C5. The NMR and UV spectroscopic studies have shown that 3'-MOE-2',5'-RNA adopts both hairpin and duplex secondary structures, which are involved in a dynamic exchange that is slow on the NMR timescale and exhibits strand and salt concentration as well as pH dependence. Unusual effect of pH over a narrow physiological range is observed for imino proton resonances with exchange broadening observed at lower pH and relatively sharp lines observed at higher pH. The solution structure of 3'-MOE-2',5'-RNA hairpin displays a unique and well-defined loop, which is stabilized by Watson-Crick A5.*U8 base pair and by n --> pi* stacking interactions of O4' lone-pair electrons of A6 and *U8 with aromatic rings of A5 and *U7, respectively. In contrast, the stem region of 3' MOE-2',5'-RNA hairpin is more flexible. Our data highlight the important feature of backbone modifications that can have pronounced effects on interstrand association of nucleic acids. PMID- 15788749 TI - Post-synthetic and site-specific modification of endocyclic nitrogen atoms of purines in DNA and its potential for biological and structural studies. AB - Site-specific modification of the N1-position of purine was explored at the nucleoside and oligomer levels. 2'-deoxyinosine was converted into an N1-2,4 dinitrophenyl derivative 2 that was readily transformed to the desired N1 substituted 2'-deoxyinosine analogues. This approach was used to develop a post synthetic method for the modification of the endocyclic N1-position of purine at the oligomer level. The phosphoramidite monomer of N1-(2,4-dinitrophenyl)-2' deoxyinosine 9 was prepared from 2'-deoxyinosine in four steps and incorporated into oligomers using an automated DNA synthesizer. The modified base, N1-(2,4 dinitrophenyl)-hypoxanthine, in synthesized oligomers, upon treatment with respective agents, was converted into corresponding N1-substituted hypoxanthines, including N1-15N-hypoxanthine, N1-methylhypoxanthine and N1-(2-aminoethyl) hypoxanthine. These modified oligomers can be easily separated and high purity oligomers obtained. Melting curve studies show the oligomer containing N1 methylhypoxanthine or N1-(2-aminoethyl)-hypoxanthine has a reduced thermostability with no particular pairing preference to either cytosine or thymine. The developed method could be adapted for the preparation of oligomers containing mutagenic N1-beta-hydroxyalkyl-hypoxanthines and the availability of the rare base-modified oligomers should offer novel tools for biological and structural studies. PMID- 15788748 TI - On the DNA cleavage mechanism of Type I restriction enzymes. AB - Although the DNA cleavage mechanism of Type I restriction-modification enzymes has been extensively studied, the mode of cleavage remains elusive. In this work, DNA ends produced by EcoKI, EcoAI and EcoR124I, members of the Type IA, IB and IC families, respectively, have been characterized by cloning and sequencing restriction products from the reactions with a plasmid DNA substrate containing a single recognition site for each enzyme. Here, we show that all three enzymes cut this substrate randomly with no preference for a particular base composition surrounding the cleavage site, producing both 5'- and 3'-overhangs of varying lengths. EcoAI preferentially generated 3'-overhangs of 2-3 nt, whereas EcoKI and EcoR124I displayed some preference for the formation of 5'-overhangs of a length of approximately 6-7 and 3-5 nt, respectively. A mutant EcoAI endonuclease assembled from wild-type and nuclease-deficient restriction subunits generated a high proportion of nicked circular DNA, whereas the wild-type enzyme catalyzed efficient cleavage of both DNA strands. We conclude that Type I restriction enzymes require two restriction subunits to introduce DNA double-strand breaks, each providing one catalytic center for phosphodiester bond hydrolysis. Possible models for DNA cleavage are discussed. PMID- 15788750 TI - Influence of the pi-pi interaction on the hydrogen bonding capacity of stacked DNA/RNA bases. AB - The interplay between aromatic stacking and hydrogen bonding in nucleobases has been investigated via high-level quantum chemical calculations. The experimentally observed stacking arrangement between consecutive bases in DNA and RNA/DNA double helices is shown to enhance their hydrogen bonding ability as opposed to gas phase optimized complexes. This phenomenon results from more repulsive electrostatic interactions as is demonstrated in a model system of cytosine stacked offset-parallel with substituted benzenes. Therefore, the H bonding capacity of the N3 and O2 atoms of cytosine increases linearly with the electrostatic repulsion between the stacked rings. The local hardness, a density functional theory-based reactivity descriptor, appears to be a key index associated with the molecular electrostatic potential (MEP) minima around H-bond accepting atoms, and is inversely proportional to the electrostatic interaction between stacked molecules. Finally, the MEP minima on surfaces around the bases in experimental structures of DNA and RNA-DNA double helices show that their hydrogen bonding capacity increases when taking more neighboring (intra-strand) stacking partners into account. PMID- 15788751 TI - Polyamines preferentially interact with bent adenine tracts in double-stranded DNA. AB - Polyamines, such as putrescine, spermidine and spermine, have indirectly been linked with the regulation of gene expression, and their concentrations are typically increased in cancer cells. Although effects on transcription factor binding to cognate DNA targets have been demonstrated, the mechanisms of the biological action of polyamines is poorly understood. Employing uranyl photo probing we now demonstrate that polyamines at submillimolar concentrations bind preferentially to bent adenine tracts in double-stranded DNA. These results provide the first clear evidence for the sequence-specific binding of polyamines to DNA, and thereby suggest a mechanism by which the cellular effects of polyamines in terms of differential gene transcriptional activity could, at least partly, be a direct consequence of sequence-specific interactions of polyamines with promoters at the DNA sequence level. PMID- 15788752 TI - The archaeal eIF2 homologue: functional properties of an ancient translation initiation factor. AB - The eukaryotic translation initiation factor 2 (eIF2) is pivotal for delivery of the initiator tRNA (tRNAi) to the ribosome. Here, we report the functional characterization of the archaeal homologue, a/eIF2. We have cloned the genes encoding the three subunits of a/eIF2 from the thermophilic archaeon Sulfolobus solfataricus, and have assayed the activities of the purified recombinant proteins in vitro. We demonstrate that the trimeric factor reconstituted from the recombinant polypeptides has properties similar to those of its eukaryal homologue: it interacts with GTP and Met-tRNAi, and stimulates binding of the latter to the small ribosomal subunit. However, the archaeal protein differs in some functional aspects from its eukaryal counterpart. In contrast to eIF2, a/eIF2 has similar affinities for GDP and GTP, and the beta-subunit does not contribute to tRNAi binding. The detailed analysis of the complete trimer and of its isolated subunits is discussed in light of the evolutionary history of the eIF2-like proteins. PMID- 15788754 TI - Weaning initiates a rapid and powerful anabolic phase in the rat maternal skeleton. AB - Maternal skeletal mineral lost during lactation is rapidly restored after weaning. The purposes of this study were to determine when increases of bone formation occur after weaning, whether the expanding osteoblast population is derived from proliferating progenitors, and to relate these skeletal changes to known endocrine events at weaning. Female rats were allowed to complete one reproductive cycle. Half of these rats were mated a second time and allowed to lactate for 20 days. The other half served as an age-matched, normal estrus cycling comparison group. One day after weaning, the dams and their comparison group were given four injections of bromodeoxyuridine (BrdU) at 8-h intervals. Indices of bone formation and the kinetics of BrdU-labeled cells were measured in lumbar vertebral cancellous bone. At 2 days after weaning, cancellous bone formation rates were substantially greater than those in the nonmated rats. Indices of bone formation more than doubled from the second to seventh day after weaning. At 25 h after the first BrdU injection in the postweaned rats, considerable numbers of labeled cells were observed on or near the bone surface, with about 17% of the osteoblast population labeled. Labeled osteoblasts peaked at 20%-24% compared with 4% in the normal estrus cycling group. Immediately following weaning, there is a profound increase in the osteoblast population in maternal cancellous bone. Many, if not most of these newly formed osteoblasts were derived from proliferating progenitors. It is possible that the endocrine milieu of lactation expands or primes the osteoprogenitor pool for this rapid anabolic phase. PMID- 15788753 TI - Sheep endogenous betaretroviruses (enJSRVs) and the hyaluronidase 2 (HYAL2) receptor in the ovine uterus and conceptus. AB - The ovine genome contains approximately 20 copies of endogenous betaretroviruses (enJSRVs) that are highly related to two exogenous oncogenic viruses, Jaagsiekte sheep retrovirus (JSRV) and Enzootic nasal tumor virus. The cellular receptor for both JSRV and the enJSRVs is hyaluronidase 2 (HYAL2). In this study, we assessed expression of enJSRVs envelope (env) and HYAL2 mRNAs in the ovine uterus and conceptus (embryo/fetus and extraembryonic membranes) throughout gestation. By reverse transcription-polymerase chain reaction analyses, enJSRVs env were found to be expressed beginning in the Day 12 conceptus, whereas HYAL2 was expressed from Day 16. HYAL2 mRNA was detected throughout gestation in the placentome but not in the endometrium, whereas enJSRVs env expression was detected throughout gestation in endometrium and placentomes. The enJSRVs env mRNA was specifically expressed in the endometrial lumenal epithelium (LE) and glandular epithelium (GE) as well as the trophoblast giant binucleate cells (BNC) and multinucleated syncytia of the placenta. HYAL2 mRNA was only detected in the BNC and multinucleated syncytial plaques of the placentome. Partial sequencing of the transcriptionally active enJSRVs from sheep endometrium, placentomes, and placenta revealed expression of many enJSRV loci. Cloning of the expressed enJSRVs env mRNA from ovine uteroplacental tissues found sequences similar to the previously identified enJS5F16 and enJS56A1 gene with an intact open reading frame, although the polypeptides they encode were not studied. Collectively, results provide further support for our hypothesis that the enJSRVs Env have been beneficial to the host and are involved in protection of the uterus from viral infection and regulators of placental morphogenesis and function. PMID- 15788755 TI - Tumor necrosis factor and vascular endothelial growth factor induce endothelial integrin repertories, regulating endovascular differentiation and apoptosis in a human extravillous trophoblast cell line. AB - Angiogenesis is crucial in human development. Extravillous trophoblast (EVT) cells mimic endothelial cells in angiogenesis during endovascular differentiation, inducing a remodeling of spiral arteries that increases blood flow toward the intravillous space. We have previously shown that tumor necrosis factor (TNF) alpha regulates expression of ITGA6 and ITGA1, which are involved in cell survival, in the human EVT cell line TCL1. To further investigate endovascular differentiation, we examined the effects of vascular endothelial growth factor (VEGF), TNF, and extracellular matrix (ECM) on TCL1 cells. Seeded on Matrigel, TCL1 cells show tube-like formation that specifically recalls morphological changes in endothelial cells. Anti-ITGAV/ITGB3 antibodies significantly reduced the size of the capillary network (P < 0.05) on Matrigel and also suppressed TNF-induced apoptosis (P < 0.05) in TCL1 cells. VEGF induced expression of ITGAV/ITGB3 subunits and protein aggregation, as in the case of TNF, which in turn, induces synthesis of VEGF in TCL1 cells. Soluble FLT1 suppressed these activities in TCL1 cells, indicating that signals involving VEGF axis are essential for endovascular differentiation. These results suggest that TNF, VEGF, and ECM collaboratively regulate EVT behavior, including cell survival and endovascular differentiation, through integrin signaling during establishment and maintenance of successful human pregnancies. PMID- 15788756 TI - Carbonic anhydrase regulate endometrial gland development in the neonatal uterus. AB - Carbonic anhydrases (CAs) are zinc metalloenyzmes that catalyze the reversible conversion of carbon dioxide to carbonic acid and are involved in respiration, calcification, acid-base balance, and formation of fluids. Transcriptional profiling of the developing neonatal mouse uterus detected expression of Car1, Car2, Car11, and Car13 between Postnatal Days (PNDs) 3 and 18. In the neonatal mouse uterus, Car2 and Car11 mRNAs were predominantly localized in endometrial epithelial and stromal cells, respectively, whereas Car13 mRNA was detected in both epithelia and stroma. CAR2 protein was detected primarily in the endometrial epithelia and from PND 3 to PND 18 in the uteri of neonatal mice. To determine whether CA regulated uterine development, neonatal mice were treated s.c. with acetazolamide, a CA inhibitor, from PND 3 to PND 18. Treatment with acetazolamide decreased CA activity in the uterus and the number of endometrial glands without apparent effects on differentiation of the stroma or myometrium. In the neonatal sheep uterus, CA2 mRNA was initially expressed at birth (PND 0) in the endometrial luminal epithelium and was predominantly expressed in the developing glandular epithelium from PND 7 to PND 56. These results support the hypothesis that CA has a functional role in endometrial gland development during postnatal uterine morphogenesis. PMID- 15788757 TI - Perturbations in the biochemical composition of fetal fluids are apparent in surviving bovine somatic cell nuclear transfer pregnancies in the first half of gestation. AB - Amniotic and allantoic fluid volumes and composition change dynamically throughout gestation. Cattle that are pregnant with somatic cell nuclear transfer (NT) fetuses show a high incidence of abnormal fluid accumulation (particularly hydrallantois) and fetal mortality from approximately midgestation. To investigate fetal fluid homeostasis in these pregnancies, Na, K, Cl, urea, creatinine, Ca, Mg, total PO(4), glucose, fructose, lactate, total protein, and osmolalities were measured in amniotic and allantoic fluids collected at Days 50, 100, and 150 of gestation from NT pregnancies and those generated by the transfer of in vitro-produced embryos or by artificial insemination. Deviations in fetal fluid composition between NT and control pregnancies were apparent after placental and fetal organ development, even when no gross morphological abnormalities were observed. Individual NT fetuses were affected to varying degrees. Elevated allantoic Na was associated with lower K and increased allantoic fluid volume or edema of the fetal membranes. Total PO(4) levels in NT allantoic and amniotic fluid were elevated at Days 100 and 150. This was not accompanied by hypophosphatemia at Day 150, suggesting that PO(4) acquisition by NT fetuses was adequate but that its readsorption by the kidneys may be impaired. Excessive NT placental weight was associated with low allantoic glucose and fructose as well as high lactate levels. However, the fructogenic ability of the NT placenta appeared to be normal. The osmolality of the fetal fluids was maintained within a narrow range, suggesting that the regulation of fluid composition, but not osmolality, was impaired in NT pregnancies. PMID- 15788758 TI - The relationship between task performance and functional magnetic resonance imaging response. AB - We compared psychophysical and functional magnetic resonance imaging (fMRI) responses within areas V1-V3 and MT+ during both a speed and a contrast discrimination task. We found that fMRI responses did not depend significantly on task in any of these areas. Moreover, responses in V1-V3 were larger than those in MT+ for both the speed and the contrast discrimination tasks across a wide range of contrasts. This pattern of results demonstrates that localizing function based on finding those regions of cortex that show greater activity to a given task-stimulus combination than to other tasks and stimuli may, under certain conditions, be misleading. However, a simple ideal observer model assuming that perceptual thresholds are dependent on neuronal population responses does successfully show that V1 has neuronal properties consistent with our subjects' contrast discrimination performance, and that MT+ has neuronal properties consistent with subjects' performance on a speed discrimination task. PMID- 15788759 TI - A diet enriched with the omega-3 fatty acid docosahexaenoic acid reduces amyloid burden in an aged Alzheimer mouse model. AB - Epidemiological studies suggest that increased intake of the omega-3 (n-3) polyunsaturated fatty acid (PUFA) docosahexaenoic acid (DHA) is associated with reduced risk of Alzheimer's disease (AD). DHA levels are lower in serum and brains of AD patients, which could result from low dietary intake and/or PUFA oxidation. Because effects of DHA on Alzheimer pathogenesis, particularly on amyloidosis, are unknown, we used the APPsw (Tg2576) transgenic mouse model to evaluate the impact of dietary DHA on amyloid precursor protein (APP) processing and amyloid burden. Aged animals (17-19 months old) were placed in one of three groups until 22.5 months of age: control (0.09% DHA), low-DHA (0%), or high-DHA (0.6%) chow. beta-Amyloid (Abeta) ELISA of the detergent-insoluble extract of cortical homogenates showed that DHA-enriched diets significantly reduced total Abeta by >70% when compared with low-DHA or control chow diets. Dietary DHA also decreased Abeta42 levels below those seen with control chow. Image analysis of brain sections with an antibody against Abeta (amino acids 1-13) revealed that overall plaque burden was significantly reduced by 40.3%, with the largest reductions (40-50%) in the hippocampus and parietal cortex. DHA modulated APP processing by decreasing both alpha- and beta-APP C-terminal fragment products and full-length APP. BACE1 (beta-secretase activity of the beta-site APP-cleaving enzyme), ApoE (apolipoprotein E), and transthyretin gene expression were unchanged with the high-DHA diet. Together, these results suggest that dietary DHA could be protective against beta-amyloid production, accumulation, and potential downstream toxicity. PMID- 15788761 TI - A model for interaural time difference sensitivity in the medial superior olive: interaction of excitatory and inhibitory synaptic inputs, channel dynamics, and cellular morphology. AB - This study reports simulations of recent physiological results from the gerbil medial superior olive (MSO) that reveal that blocking glycinergic inhibition can shift the tuning for the interaural time difference (ITD) of the cell (Brand et al., 2002). Our simulations indicate that the model proposed in the study by Brand et al. (2002) requires precisely timed, short-duration inhibition with temporal accuracy exceeding that described in the auditory system. An alternative model is proposed that incorporates two anatomic observations in the MSO: (1) the axon arises from the dendrite that receives ipsilateral inputs; and (2) inhibitory synapses are located primarily on the soma in adult animals. When the inhibitory currents are activated or blocked, the model cell successfully simulates experimentally observed shifts in the best ITD. The asymmetrical cell structure allows an imbalance between the ipsilateral and contralateral excitatory inputs and shifts the ITD curve such that the best ITD is not at zero. Fine adjustment of the best ITD is achieved by the interplay of somatic sodium currents and synaptic inhibitory currents. The shift of the best ITD in the model is limited to approximately 0.2 ms, which is behaviorally significant with respect to ITDs encountered in perceptual tasks. The model suggests a mechanism for dynamically "fine-tuning" the ITD sensitivity of MSO cells by the opponency between depolarizing sodium currents and hyperpolarizing inhibitory currents. PMID- 15788760 TI - Requirement of nicotinic acetylcholine receptor subunit beta2 in the maintenance of spiral ganglion neurons during aging. AB - Age-related hearing loss (presbycusis) is a major health concern for the elderly. Loss of spiral ganglion neurons (SGNs), the primary sensory relay of the auditory system, is associated consistently with presbycusis. The causative molecular events responsible for age-related loss of SGNs are unknown. Recent reports directly link age-related neuronal loss in cerebral cortex with the loss of high affinity nicotine acetylcholine receptors (nAChRs). In cochlea, cholinergic synapses are made by olivocochlear efferent fibers on the outer hair cells that express alpha9 nAChR subunits and on the peripheral projections of SGNs that express alpha2, alpha4-7, and beta2-3 nAChR subunits. A significantly decreased expression of the beta2 nAChR subunit in SGNs was found specifically in mice susceptible to presbycusis. Furthermore, mice lacking the beta2 nAChR subunit (beta2-/-), but not mice lacking the alpha5 nAChR subunit (alpha5-/-), have dramatic hearing loss and significant reduction in the number of SGNs. Our findings clearly established a requirement for beta2 nAChR subunit in the maintenance of SGNs during aging. PMID- 15788762 TI - Deficits in experience-dependent cortical plasticity and sensory-discrimination learning in presymptomatic Huntington's disease mice. AB - Huntington's disease (HD) is one of a group of neurodegenerative diseases caused by an expanded trinucleotide (CAG) repeat coding for an extended polyglutamine tract. The disease is inherited in an autosomal dominant manner, with onset of motor, cognitive, and psychiatric symptoms typically occurring in midlife, followed by unremitting progression and eventual death. We report here that motor presymptomatic R6/1 HD mice show a severe impairment of somatosensory discrimination learning ability in a behavioral task that depends heavily on the barrel cortex. In parallel, there are deficits in barrel-cortex plasticity after a somatosensory whisker-deprivation paradigm. The present study demonstrates deficits in neocortical plasticity correlated with a specific learning impairment involving the same neocortical area, a finding that provides new insight into the cellular basis of early cognitive deficits in HD. PMID- 15788763 TI - Distinct roles of calcineurin-nuclear factor of activated T-cells and protein kinase A-cAMP response element-binding protein signaling in presynaptic differentiation. AB - Synaptic vesicle accumulation and morphological changes are characteristic features of axon terminal differentiation during synaptogenesis. To investigate the regulatory mechanism that orchestrates synaptic molecules to form mature presynaptic terminals, we visualized a single axon terminal of zebrafish olfactory sensory neurons in vivo and examined the effects of the neuron-specific gene manipulations on the axon terminal differentiation. Synaptic vesicles visualized with vesicle-associated membrane protein 2 (VAMP2)-enhanced green fluorescent protein (EGFP) fusion protein gradually accumulated in axon terminals, whereas the axon terminals visualized with GAP43 fused with EGFP remodeled from complex shapes with filopodia to simple shapes without filopodia from 50 h postfertilization (hpf) to 84 hpf. Expression of dominant-negative protein kinase A (PKA) or cAMP response element-binding protein (CREB) suppressed the VAMP2-EGFP punctum formation in axon terminals during synaptogenesis. Consistently, constitutively active PKA or CREB stimulated VAMP2-EGFP puncta formation. On the other hand, cyclosporine A treatment or suppression of nuclear factor of activated T cells (NFAT) activation prevented the axon terminal remodeling from complex to simple shapes during synaptogenesis. Consistently, expression of constitutively active calcineurin accelerated the axon terminal remodeling. These results suggest that calcineurin-NFAT signaling regulates axon terminal remodeling, and PKA-CREB signaling controls synaptic vesicle accumulation. PMID- 15788764 TI - Early presynaptic and late postsynaptic components contribute independently to brain-derived neurotrophic factor-induced synaptic plasticity. AB - Trophin-induced synaptic plasticity consists of both presynaptic and postsynaptic processes. The potential interdependence of these mechanisms and their temporal relationships are undefined. The synaptic vesicle protein Rab3A is required for the early, initial 10 min phase but not for the later phase of BDNF-enhanced transmission. We now examine the temporal distinction and mechanistic relationships between these phases of BDNF action. Rab3A mutant cells did not exhibit increased miniature EPSC frequency in response to BDNF in cell culture, indicating an absence of the presynaptic component. In contrast, BDNF enhanced postsynaptic glutamate-induced current in the mutant neurons as in the wild type, indicating that the postsynaptic component of the response was intact. Finally, the postsynaptic NMDA receptor subunit NR2B was phosphorylated at Tyr1472 by BDNF in Rab3A knock-outs, as shown previously in wild type. Our results are the first to demonstrate that presynaptic and postsynaptic components of BDNF-enhanced synaptic activity are independent and temporally distinct. PMID- 15788765 TI - The retrograde spread of synaptic potentials and recruitment of presynaptic inputs. AB - Lateral excitation is a mechanism for amplifying coordinated input to postsynaptic neurons that has been described recently in several species. Here, we describe how a postsynaptic neuron, the lateral giant (LG) escape command neuron, enhances lateral excitation among its presynaptic mechanosensory afferents in the crayfish tailfan. A lateral excitatory network exists among electrically coupled tailfan primary afferents, mediated through central electrical synapses. EPSPs elicited in LG dendrites as a result of mechanosensory stimulation spread antidromically back through electrical junctions to unstimulated afferents, summate with EPSPs elicited through direct afferent-to afferent connections, and contribute to recruitment of these afferents. Antidromic potentials are larger if the afferent is closer to the initial input on LG dendrites, which could create a spatial filtering mechanism within the network. This pathway also broadens the temporal window over which lateral excitation can occur, because of the delay required for EPSPs to spread through the large LG dendrites. The delay allows subthreshold inputs to the LG to have a priming effect on the lateral excitatory network and lowers the threshold of the network in response to a second, short-latency stimulus. Retrograde communication within neuronal pathways has been described in a number of vertebrate and invertebrate species. A mechanism of antidromic passage of depolarizing current from a neuron to its presynaptic afferents, similar to that described here in an invertebrate, is also present in a vertebrate (fish). This raises the possibility that short-term retrograde modulation of presynaptic elements through electrical junctions may be common. PMID- 15788766 TI - Mechanisms of transport and exocytosis of dense-core granules containing tissue plasminogen activator in developing hippocampal neurons. AB - Dense-core granules (DCGs) are organelles found in specialized secretory cells, including neuroendocrine cells and neurons. Neuronal DCGs facilitate many critical processes, including the transport and secretion of proteins involved in learning, and yet their transport and exocytosis are poorly understood. We have used wide-field and total internal reflection fluorescence microscopy, in conjunction with transport theory, to visualize the transport and exocytosis of DCGs containing a tissue plasminogen activator-green fluorescent protein hybrid in cell bodies, neurites, and growth cones of developing hippocampal neurons and to quantify the roles that diffusion, directed motion, and immobility play in these processes. Our results demonstrate that shorter-ranged transport of DCGs near sites of exocytosis in hippocampal neurons and neuroendocrine cells differs markedly. Specifically, the immobile fraction of DCGs within growth cones and near the plasma membrane of hippocampal neurons is small and relatively unaltered by actin disruption, unlike in neuroendocrine cells. Moreover, transport of DCGs in these domains of hippocampal neurons is unusually heterogeneous, being significantly rapid and directed as well as slow and diffusive. Our results also demonstrate that exocytosis is preceded by substantial movement and heterogeneous transport; this movement may facilitate delivery of DCG cargo in hippocampal neurons, given the relatively low abundance of neuronal DCGs. In addition, the extensive mobility of DCGs in hippocampal neurons argues strongly against the hypothesis that cortical actin is a major barrier to membrane-proximal DCGs in these cells. Instead, our results suggest that extended release of DCG cargo from hippocampal neurons arises from heterogeneity in DCG mobility. PMID- 15788767 TI - Visualization of synaptic Ca2+ /calmodulin-dependent protein kinase II activity in living neurons. AB - Ca2+/calmodulin-dependent protein kinase II (CaMKII) is highly enriched in excitatory synapses in the CNS and critically involved in synaptic plasticity, learning, and memory. However, the precise temporal and spatial regulation of CaMKII activity in living cells has not been well described, because of a lack of specific methods. We tried to address this by optically detecting the conformational change in CaMKII during activation using fluorescence resonance energy transfer (FRET). The engineered FRET probe Camuialpha detects calmodulin binding and autophosphorylation at threonine 286 that renders the enzyme constitutively active. In combination with two-photon microscopy, we demonstrate that Camuialpha can be used to observe temporal and spatial regulation of CaMKII activity in living neurons. PMID- 15788768 TI - Calcium sensitivity of neurotransmitter release differs at phasic and tonic synapses. AB - The efficacy of synaptic transmission varies greatly among synaptic contacts. We have explored the origins of differences between phasic and tonic crustacean neuromuscular junctions. Synaptic boutons of a phasic motor neuron release three orders of magnitude more quanta to a single action potential and show strong depression to a train, whereas tonic synapses are nearly unresponsive to single action potentials and display an immense facilitation. Phasic and tonic synapses display a similar nonlinear dependence on extracellular [Ca2+]. We imposed similar spatially uniform intracellular [Ca2+] ([Ca2+]i) steps in phasic and tonic synapses by photolysis of presynaptic caged calcium. [Ca2+]i was measured fluorometrically while transmitter release was monitored electrophysiologically from single boutons in which the [Ca2+]i was elevated. Phasic synapses released the readily releasable pool (RRP) of vesicles at a much higher rate and with a shorter delay than did tonic synapses. Comparison of several kinetic models of molecular events showed that a difference in Ca2+-sensitive priming of vesicles in the RRP combined with a revision of the kinetic Ca2+-binding sequence to the secretory trigger produced the best fit to the markedly different responses to Ca2+ steps and action potentials and of the characteristic features of synaptic plasticity in phasic and tonic synapses. The results reveal processes underlying one aspect of synaptic diversity that may also regulate changes in synaptic strength during development and learning and memory formation. PMID- 15788769 TI - Selective blockade of the capsaicin receptor TRPV1 attenuates bone cancer pain. AB - Cancer colonization of bone leads to the activation of osteoclasts, thereby producing local tissue acidosis and bone resorption. This process may contribute to the generation of both ongoing and movement-evoked pain, resulting from the activation of sensory neurons that detect noxious stimuli (nociceptors). The capsaicin receptor TRPV1 (transient receptor potential vanilloid subtype 1) is a cation channel expressed by nociceptors that detects multiple pain-producing stimuli, including noxious heat and extracellular protons, raising the possibility that it is an important mediator of bone cancer pain via its capacity to detect osteoclast- and tumor-mediated tissue acidosis. Here, we show that TRPV1 is present on sensory neuron fibers that innervate the mouse femur and that, in an in vivo model of bone cancer pain, acute or chronic administration of a TRPV1 antagonist or disruption of the TRPV1 gene results in a significant attenuation of both ongoing and movement-evoked nocifensive behaviors. Administration of the antagonist had similar efficacy in reducing early, moderate, and severe pain-related responses, suggesting that TRPV1 may be a novel target for pharmacological treatment of chronic pain states associated with bone cancer metastasis. PMID- 15788770 TI - Deleted in colorectal cancer binding netrin-1 mediates cell substrate adhesion and recruits Cdc42, Rac1, Pak1, and N-WASP into an intracellular signaling complex that promotes growth cone expansion. AB - Extracellular cues direct axon extension by regulating growth cone morphology. The netrin-1 receptor deleted in colorectal cancer (DCC) is required for commissural axon extension to the floor plate in the embryonic spinal cord. Here we demonstrate that challenging embryonic rat spinal commissural neurons with netrin-1, either in solution or as a substrate, causes DCC-dependent increases in growth cone surface area and filopodia number, which we term growth cone expansion. We provide evidence that DCC influences growth cone morphology by at least two mechanisms. First, DCC mediates an adhesive interaction with substrate bound netrin-1. Second, netrin-1 binding to DCC recruits an intracellular signaling complex that directs the organization of actin. We show that netrin-1 induced growth cone expansion requires Cdc42 (cell division cycle 42), Rac1 (Ras related C3 botulinum toxin substrate 1), Pak1 (p21-activated kinase), and N-WASP (neuronal Wiskott-Aldrich syndrome protein) and that the application of netrin-1 rapidly activates Cdc42, Rac1, and Pak1. Furthermore, netrin-1 recruits Cdc42, Rac1, Pak1, and N-WASP into a complex with the intracellular domain of DCC and Nck1. These findings suggest that DCC influences growth cone morphology by acting both as a transmembrane bridge that links extracellular netrin-1 to the actin cytoskeleton and as the core of a protein complex that directs the organization of actin. PMID- 15788771 TI - Disruption of ephrin signaling associates with disordered axophilic migration of the gonadotropin-releasing hormone neurons. AB - Ephrin signaling is involved in repulsive and attractive interactions mediating axon guidance and cell-boundary formation in the developing nervous system. As a result of a fortuitous transgene integration event, we have identified here a potential role for EphA5 in the axophilic migration of gonadotropin-releasing hormone (GnRH) neurons from the nasal placode into the brain along ephrin expressing vomeronasal axons. Transgene integration in the GNR23 mouse line resulted in a 26 kb deletion in chromosome 5, approximately 67 kb 3' to Epha5. This induced a profound, region-specific upregulation of EphA5 mRNA and protein expression in the developing mouse brain. The GnRH neurons in GNR23 mice overexpressed EphA5 from embryonic day 11, whereas ephrin A3 and A5 mRNA levels in olfactory neurons were unchanged. The GnRH neurons were found to be slow in commencing their migration from the olfactory placode and also to form abnormal clusters of cells on the olfactory axons, prohibiting their migration out of the nose. As a result, adult hemizygous mice had only 40% of the normal complement of GnRH neurons in the brain, whereas homozygous mice had <15%. This resulted in infertility in adult female homozygous GNR23 mice, suggesting that some cases of human hypogonadotropic hypogonadism may result from ephrin-related mutations. These data provide evidence for a role of EphA-ephrin signaling in the axophilic migration of the GnRH neurons during embryogenesis. PMID- 15788772 TI - Emotional autobiographical memories in amnesic patients with medial temporal lobe damage. AB - The recollection of emotional autobiographical memories has received little attention in patients with memory disorders. Here, we addressed this topic in amnesic patients with damage to the hippocampus (HC group; n = 8) or the hippocampus, amygdala, and surrounding cortices (HC+ group; n = 2). These patients were asked to recollect emotional events from their lives. HC patients produced recollections that were strikingly similar to those of brain-damaged (n = 10) and healthy (n = 25) comparison participants, in terms of both quantity and quality. In contrast, HC+ patients produced a lower proportion of unpleasant memories compared with the other participants. Specifically, the ratings and words used to describe recollections in the HC+ patients were more affectively positive. All groups produced more memories from between 10 and 30 years of age (the so-called autobiographical memory "bump") compared with other time periods in their lives. These results suggest that structures surrounding the hippocampus, but not the hippocampus itself, may be necessary for the recollection of highly emotional, unpleasant autobiographical memories. The amygdala and surrounding cortices of the medial temporal lobe may be a necessary component in the neural circuitry necessary for vivid recollection of unpleasant emotional events. PMID- 15788773 TI - Anatomy of spatial attention: insights from perfusion imaging and hemispatial neglect in acute stroke. AB - The site of lesion responsible for left hemispatial neglect after stroke has been intensely debated recently. Some studies provide evidence that right angular lesions are most likely to cause left neglect, whereas others indicate that right superior temporal lesions are most likely to cause neglect. We examine two potential accounts of the conflicting results: (1) neglect could result from cortical dysfunction beyond the structural lesion in some studies; and (2) different forms of neglect with separate neural correlates have been included in different proportions in separate studies. To evaluate these proposals, we studied 50 patients with acute right subcortical infarcts using tests of hemispatial neglect and magnetic resonance diffusion-weighted and perfusion weighted imaging performed within 48 h of onset of symptoms. Left "allocentric" neglect (errors on the left sides of individual stimuli, regardless of location with respect to the viewer) was most strongly associated with hypoperfusion of right superior temporal gyrus (Fisher's exact test; p < 0.0001), whereas left "egocentric" neglect (errors on the left of the viewer) was most strongly associated with hypoperfusion of the right angular gyrus (p < 0.0001). Patients without cortical hypoperfusion showed no hemispatial neglect. Because the patients did not have cortical infarcts, our data show that neglect can be caused by hypoperfused dysfunctional tissue not detectable by structural magnetic resonance imaging. Moreover, different forms of neglect were associated with different sites of cortical hypoperfusion. Results help explain conflicting results in the literature and contribute to the understanding of spatial attention and representation in the human brain. PMID- 15788774 TI - Phosphorylation-dependent low-frequency depression at phasic synapses of a crayfish motoneuron. AB - Extremes in presynaptic differentiation can be studied at the crayfish leg extensor muscle where, on the same muscle fiber, one motoneuron makes "phasic" depressing synapses that have a high probability of neurotransmitter release and another motoneuron makes "tonic," low-probability, facilitating synapses. The large motor axons permit intracellular access to presynaptic sites. We examined the role of phosphorylation during low-frequency depression (LFD) in the relatively little studied phasic synapses. LFD occurs with stimulation at 0.2 Hz and develops with time constants of 4 and 105 min to reach >50% depression of transmitter release in 60 min similar to long-term depression in mammals. LFD is not associated with changes in postsynaptic sensitivity to transmitter and thus is a presynaptic event, although it is not accompanied by changes in the presynaptic action potential. Blockade of protein kinases accelerated the slow phase of LFD, but stimulation of kinases reduced depression. Blockade of protein phosphatases 1A/2A reversed the slow phase. When calcineurin was inhibited, both phases of LFD were abolished, and facilitation occurred instead. Immunostaining showed calcineurin-like immunoreactivity in synaptic terminals. Recovery from LFD occurred in approximately 1 h if stimulation frequency was reduced to 0.0016 Hz. Recovery was blocked by kinase inhibition. This study shows that phosphorylation dependent mechanisms are involved in LFD and suggests that exocytosis is controlled by conditions that shift the balance between phosphorylated and unphosphorylated substrates. The shift can occur by alteration in the relative activities of protein kinases and phosphatases. PMID- 15788775 TI - Intrinsic musculoskeletal properties stabilize wiping movements in the spinalized frog. AB - The mechanical stability properties of hindlimb-hindlimb wiping movements of the spinalized frog were examined. One hindlimb, the wiping limb, was implanted with 12 electromyographic (EMG) electrodes and attached to a robot that both recorded its trajectory and applied brief force perturbations. Cutaneous electrical stimulation was applied to the other hindlimb, the target limb, to evoke the hindlimb-hindlimb wiping reflex. Kinematic and EMG data from both unperturbed trials and trials in which a phasic perturbation was applied were collected from each spinalized frog. In the perturbed behaviors, we found that the initially large displacement attributable to the perturbation was compensated such that the final position was statistically indistinguishable from the unperturbed final position in all of the frogs, thus indicating the dynamic stability of these movements. This stability was robust to the range of perturbation amplitudes and nominal kinematic variation observed in this study. In addition, we investigated the extent to which intrinsic viscoelastic properties of the limb and proprioceptive feedback play a role in stabilizing the movements. No significant changes were seen in the EMGs after the perturbation. Furthermore, deafferentation of the wiping limb did not significantly affect the stability of the wiping reflex. Thus, we found that the intrinsic viscoelastic properties of the hindlimb conferred robust stability properties to the hindlimb-hindlimb wiping behavior. This stability mechanism may simplify the control required by the frog spinal motor systems to produce successful movements in an unpredictable and varying environment. PMID- 15788776 TI - Induction of delta-opioid receptor function in the midbrain after chronic morphine treatment. AB - Delta-opioid receptor (DOPr) activation fails to produce cellular physiological responses in many brain regions, including the periaqueductal gray (PAG), despite neural expression of high densities of the receptor. Previous histochemical studies have demonstrated that a variety of stimuli, including chronic morphine treatment, induce the translocation of DOPr from intracellular pools to the surface membrane of CNS neurons. PAG neurons in slices taken from untreated mice exhibited mu-opioid receptor (MOPr) but not DOPr-mediated presynaptic inhibition of GABAergic synaptic currents. In contrast, after 5-6 d of chronic morphine treatment, DOPr stimulation inhibited synaptic GABA release onto most neurons. Shorter exposure to morphine in vitro (upto 4 h) or in vivo (18 h) did not induce functional DOPr responses. DOPr-mediated presynaptic inhibition could not be induced in slices from untreated animals by increasing synaptic activity in vitro using high extracellular potassium concentrations or activation of protein kinase A. Induction of functional DOPr signaling by chronic morphine required MOPr expression, because no DOPr receptor responses were observed in MOPr knock-out mice. DOPr agonists also had no effect on miniature IPSCs in beta-arrestin-2 knock-out mice after chronic morphine. These results suggest that induction of DOPr-mediated actions in PAG by chronic morphine requires prolonged MOPr stimulation and expression of beta-arrestin-2. PMID- 15788777 TI - An essential Drosophila glutamate receptor subunit that functions in both central neuropil and neuromuscular junction. AB - A Drosophila forward genetic screen for mutants with defective synaptic development identified bad reception (brec). Homozygous brec mutants are embryonic lethal, paralyzed, and show no detectable synaptic transmission at the glutamatergic neuromuscular junction (NMJ). Genetic mapping, complementation tests, and genomic sequencing show that brec mutations disrupt a previously uncharacterized ionotropic glutamate receptor subunit, named here "GluRIID." GluRIID is expressed in the postsynaptic domain of the NMJ, as well as widely throughout the synaptic neuropil of the CNS. In the NMJ of null brec mutants, all known glutamate receptor subunits are undetectable by immunocytochemistry, and all functional glutamate receptors are eliminated. Thus, we conclude that GluRIID is essential for the assembly and/or stabilization of glutamate receptors in the NMJ. In null brec mutant embryos, the frequency of periodic excitatory currents in motor neurons is significantly reduced, demonstrating that CNS motor pattern activity is regulated by GluRIID. Although synaptic development and molecular differentiation appear otherwise unperturbed in null mutants, viable hypomorphic brec mutants display dramatically undergrown NMJs by the end of larval development, suggesting that GluRIID-dependent central pattern activity regulates peripheral synaptic growth. These studies reveal GluRIID as a newly identified glutamate receptor subunit that is essential for glutamate receptor assembly/stabilization in the peripheral NMJ and required for properly patterned motor output in the CNS. PMID- 15788778 TI - Four different subunits are essential for expressing the synaptic glutamate receptor at neuromuscular junctions of Drosophila. AB - Three ionotropic glutamate receptor subunits, designated GluRIIA, GluRIIB, and GluRIII, have been identified at neuromuscular junctions of Drosophila. Whereas GluRIIA and GluRIIB are redundant for viability, it was shown recently that GluRIII is essential for both the synaptic localization of GluRIIA and GluRIIB and the viability of Drosophila. Here we identify a fourth and a fifth subunit expressed in the neuromuscular system, which we name GluRIID and GluRIIE. Both new subunits we show to be necessary for survival. Moreover, both GluRIID and GluRIIE are required for the synaptic expression of all other glutamate receptor subunits. All five subunits are interdependent for receptor function, synaptic receptor expression, and viability. This indicates that synaptic glutamate receptors incorporate the GluRIII, GluRIID, and GluRIIE subunit together with either GluRIIA or GluRIIB at the Drosophila neuromuscular junction. At this widely used model synapse, the assembly of four different subunits to form an individual glutamate receptor channel may thus be obligatory. This study opens the way for a further characterization of in vivo glutamate receptor assembly and trafficking using the efficient genetics of Drosophila. PMID- 15788779 TI - Differential regulation of AMPA receptor and GABA receptor trafficking by tumor necrosis factor-alpha. AB - The proinflammatory cytokine tumor necrosis factor-alpha (TNFalpha) causes a rapid exocytosis of AMPA receptors in hippocampal pyramidal cells and is constitutively required for the maintenance of normal surface expression of AMPA receptors. Here we demonstrate that TNFalpha acts on neuronal TNFR1 receptors to preferentially exocytose glutamate receptor 2-lacking AMPA receptors through a phosphatidylinositol 3 kinase-dependent process. This increases excitatory synaptic strength while changing the molecular stoichiometry of synaptic AMPA receptors. Conversely, TNFalpha causes an endocytosis of GABA(A) receptors, resulting in fewer surface GABA(A) receptors and a decrease in inhibitory synaptic strength. These results suggest that TNFalpha can regulate neuronal circuit homeostasis in a manner that may exacerbate excitotoxic damage resulting from neuronal insults. PMID- 15788781 TI - GABA transporter deficiency causes tremor, ataxia, nervousness, and increased GABA-induced tonic conductance in cerebellum. AB - GABA transporter subtype 1 (GAT1) knock-out (KO) mice display normal reproduction and life span but have reduced body weight (female, -10%; male, -20%) and higher body temperature fluctuations in the 0.2-1.5/h frequency range. Mouse GAT1 (mGAT1) KO mice exhibit motor disorders, including gait abnormality, constant 25 32 Hz tremor, which is aggravated by flunitrazepam, reduced rotarod performance, and reduced locomotor activity in the home cage. Open-field tests show delayed exploratory activity, reduced rearing, and reduced visits to the central area, with no change in the total distance traveled. The mGAT1 KO mice display no difference in acoustic startle response but exhibit a deficiency in prepulse inhibition. These open-field and prepulse inhibition results suggest that the mGAT1 KO mice display mild anxiety or nervousness. The compromised GABA uptake in mGAT1 KO mice results in an increased GABA(A) receptor-mediated tonic conductance in both cerebellar granule and Purkinje cells. The reduced rate of GABA clearance from the synaptic cleft is probably responsible for the slower decay of spontaneous IPSCs in cerebellar granule cells. There is little or no compensatory change in other proteins or structures related to GABA transmission in the mGAT1 KO mice, including GAT1-independent GABA uptake, number of GABAergic interneurons, and GABA(A)-, vesicular GABA transporter-, GAD65-, and GAT3 immunoreactive structures in cerebellum or hippocampus. Therefore, the excessive extracellular GABA present in mGAT1 KO mice results in behaviors that partially phenocopy the clinical side effects of tiagabine, suggesting that these side effects are inherent to a therapeutic strategy that targets the widely expressed GAT1 transporter system. PMID- 15788780 TI - In vivo activation of a mutant mu-opioid receptor by naltrexone produces a potent analgesic effect but no tolerance: role of mu-receptor activation and delta receptor blockade in morphine tolerance. AB - Opioid analgesics are the standard therapeutic agents for the treatment of pain, but their prolonged use is limited because of the development of tolerance and dependence. Recently, we reported the development of a mu-opioid receptor knock in (KI) mouse in which the mu-opioid receptor was replaced by a mutant receptor (S196A) using a homologous recombination gene-targeting strategy. In these animals, the opioid antagonist naltrexone elicited antinociceptive effects similar to those of partial agonists acting in wild-type (WT) mice; however, development of tolerance and physical dependence were greatly reduced. In this study, we test the hypothesis that the failure of naltrexone to produce tolerance in these KI mice is attributable to its simultaneous inhibition of delta-opioid receptors and activation of mu-opioid receptors. Simultaneous implantation of a morphine pellet and continuous infusion of the delta-opioid receptor antagonist naltrindole prevented tolerance development to morphine in both WT and KI animals. Moreover, administration of SNC-80 [(+)-4-[(alphaR)-alpha-((2S,5R)-4 allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide], a delta agonist, in the naltrexone-pelleted KI animals resulted in a dose-dependent induction in tolerance development to both morphine- and naltrexone-induced analgesia. We conclude that although simultaneous activation of both mu- and delta-opioid receptors results in tolerance development, mu-opioid receptor activation in conjunction with delta-opioid receptor blockade significantly attenuates the development of tolerance. PMID- 15788782 TI - Differences in transmission properties and susceptibility to long-term depression reveal functional specialization of ascending axon and parallel fiber synapses to Purkinje cells. AB - An understanding of the patterns of mossy fiber transmission to Purkinje cells, via granule cell axons, is fundamental to models of cerebellar cortical signaling and processing. Early theories assumed that mossy fiber input is widely disseminated across the cerebellar cortex along beams of parallel fibers, which spread for several millimeters across the cerebellar cortex. Direct evidence for this has, however, proved controversial, leading to the development of an alternative hypothesis that mossy fiber inputs to the cerebral cortex are in fact vertically organized such that the ascending segment of the granule axon carries a greater synaptic weight than the parallel fiber segment. Here, we report that ascending axon synapses are selectively resistant to cerebellar long-term depression and that they release transmitter with higher mean release probabilities and mean quantal amplitudes than parallel fiber synapses. This novel specialization of synapses formed by different segments of the same axon not only explains the reported patterns of granule cell--> Purkinje cell transmission across the cerebellar cortex but also reveals an additional level of functionality and complexity of cerebellar processing. Consequently, ascending axon synapses represent a new element of cortical signal processing that should be distinguished from parallel fiber synapses in future experimental and theoretical studies of cerebellar function. PMID- 15788784 TI - Global harmonization of hemoglobin A1c. PMID- 15788785 TI - Issues in immunoassay standardization: the ARCHITECT Folate model for intermethod harmonization. PMID- 15788783 TI - Rapid motif-based prediction of circular permutations in multi-domain proteins. AB - MOTIVATION: Rearrangements of protein domains and motifs such as swaps and circular permutations (CPs) can produce erroneous results in searching sequence databases when using traditional methods based on linear sequence alignments. Circular permutations are also of biological relevance because they can help to better understand both protein evolution and functionality. RESULTS: We have developed an algorithm, RASPODOM, which is based on the classical recursive alignment scheme. Sequences are represented as strings of domains taken from precompiled resources of domain (motif) databases such as ProDom. The algorithm works several orders of magnitude faster than a reimplementation of the existing CP detection algorithm working on strings of amino acids, produces virtually no false positives and allows the discrimination of true CPs from 'intermediate' CPs (iCPs). Several true CPs which have not been reported in literature so far could be identified from Swiss-Prot/TrEMBL within minutes. PMID- 15788786 TI - Biochip for K-ras mutation screening in ovarian cancer. PMID- 15788787 TI - Measurement of cystine in urine by liquid chromatography-tandem mass spectrometry. PMID- 15788788 TI - False-negative result for cocaine metabolites on a lateral-flow drug test slide corrected by dilution. PMID- 15788789 TI - Are glucokinase mutations associated with low triglycerides? PMID- 15788790 TI - Precision of computed spectrophotometric scans for heme pigments in subarachnoid hemorrhage makes interpretation reliable. PMID- 15788791 TI - Beta blocker. PMID- 15788792 TI - Age-related neurodegenerative changes and how they affect the gut. AB - The enteric nervous system (ENS) is the division of the autonomic nervous system that regulates gastrointestinal (GI) function. Although large numbers of enteric neurons may be lost with age, the GI tract remains surprisingly functional. Exceptions to this generality include swallowing disorders and reduced colonic motility in the elderly. Evidence of age-related neurodegenerative changes in structure and function of the ENS is briefly reviewed in this Perspective. PMID- 15788794 TI - Counting the cost of violence in the Middle East. PMID- 15788795 TI - Public health challenges in a globalizing world. PMID- 15788796 TI - Health promotion in hospitals--a strategy to improve quality in health care. PMID- 15788797 TI - A prospective population based study of childhood injuries: the Velestino town study. AB - BACKGROUND: Unintentional injuries in childhood constitute a significant public health problem. Our purpose is to estimate the incidence of identifiable unintentional childhood injuries of any type and severity, and to document risk factors of non-transient nature. METHODS: We have undertaken a prospective population-based investigation in a Greek town with a population of 748 children (0-14 years old). All identifiable injuries have been monitored during a twelve month period through information provided by the health care outlets or educational institutions as well as the police station and the regional hospital. RESULTS: The overall incidence was 28.2 per 100 person-years (95% confidence interval from 24.4 to 32.0), whereas the incidence of injuries with Hopkins Injury Severity Score equal to or higher than four was 6.3 with 95% confidence interval 4.5 to 8.1. The incidence of total injuries was higher among boys than among girls (p<0.01) and the gender difference was particularly evident among older children. Almost half of the injuries were due to falls and more than 20% were due to cutting. Children of younger and less educated parents have higher risk for injury and children from families with more injuries were more likely to be injured themselves. There was no evidence that somatometric characteristics were associated with injury risk. CONCLUSION: The incidence of unintentional childhood injuries is high and represents a considerable health burden. Family related variables are important risk factors for childhood injuries, whereas somatometric characteristics play a minimal role. KEY POINTS: Incidence and risk factors of all injuries in a population-based study among children. About 28 per 100 children got injured over a period of one year. There is evidence that younger paternal age and lower education may be associated with increased injury risk. Family related variables seem to be important risk factors for childhood injuries, whereas somatometric characteristics play minimal role. PMID- 15788798 TI - Social inequalities in perinatal and infant mortality in the northern region of Belgium (the Flanders). AB - BACKGROUND: The study was intended to analyse the independent effect of some facets of the socio-economic status of both parents on perinatal, neonatal and post-neonatal mortality in the northern region of Belgium (the Flanders). METHOD: Perinatal data collected by the Study Centre for Perinatal Epidemiology were linked with socio-economic data collected by the district council. Mothers aged > or =25 years are included in the study. RESULTS: 50796 births were analysed. 452 infants died either before birth or during the first year of life. 52% of the foeto-infantile mortality occurred before birth and 57% of the infant mortality in the first week of life. The educational level was strongly related to foetal (p<0.001) and, to a lesser degree, to early-neonatal mortality (p=0.001). Employment did not correlate with any mortality item. Except for foetal mortality, the strongest correlation was always observed for maternal rather than paternal social items. In a logistic regression model, foetal mortality, perinatal mortality and infantile mortality remained strongly correlated with the educational level of the mother. Infant mortality beyond the first week of life was not correlated with any aspect of the social status of the parents. CONCLUSIONS: The educational level of the mother is the single most important determinant of infantile mortality in the Flanders representing the totality of hospital births by mothers aged > or =25 years in 1999 in the Flanders. KEY POINTS: STUDY QUESTION: Does education, profession and actual employment of both parents, independently operates discrimination in the outcome of pregnancy up to one year? Results: Maternal education is the only significant and independent determinant of foetal as well as neonatal and foeto-infantile mortality. Results: The status of the mother is by far more important than that of the father in determining the outcome of pregnancy. PMID- 15788799 TI - Health expectancy in Denmark, 1987-2000. AB - BACKGROUND: While life expectancy quantifies average length of life, health expectancy represents the average lifetime in different health states and offers the possibility to evaluate quality of life with respect to health. The purpose of the study was to estimate changes in health expectancy in Denmark from 1987 to 2000 and to assess theories about the relation between increased total lifetime and lifetime in various health states. METHODS: Data on health status derived from the Danish Health Interview Surveys carried out in 1987, 1991, 1994 and 2000 were combined with life-table data. Expected lifetime in selfrated good health, life expectancy without longstanding illness and disabilityfree life expectancy were estimated by Sullivan's method. RESULTS: In 1987, the life expectancy of a 65-year-old man was 14.1 years, 8.9 years of which were expected to be disabilityfree. In 2000, life expectancy had increased to 15.0 years, 11.3 years of which were disabilityfree. Thus, life expectancy had increased by 0.9 years, whereas disabilityfree life expectancy had increased by 2.4 years. Among 65-year old women, life expectancy had increased by 0.2 years and disabilityfree life expectancy by 1.1 years. Expected lifetime in selfrated good health had also improved, but the trend in life expectancy without longstanding illness went in the opposite direction, and expected lifetime with longstanding illness had increased. CONCLUSION: The recent rise in life expectancy in Denmark after many years of stagnation appears to be accompanied by generally improved health status among the elderly, but health expectancy trends depend on the health indicator chosen. KEY POINTS: Health expectancy expresses average lifetime in various states of health. The study examines changes in expected lifetime in selfrated good health, lifetime without longstanding illness and lifetime without longterm disability. Among 65-year-olds the percentage of disabilityfree life expectancy increased from 63.4% to 74.9% for men and from 55.6% to 61.0% for women between 1987 and 2000. Health status among elderly Danes has apparently improved, but secular trends in health expectancy depend on the choice of health indicator. PMID- 15788800 TI - A preliminary evaluation of a health monitoring programme in Hungary. AB - BACKGROUND: In 1998 a joint initiative of the Hungarian School of Public Health and the National Public Health Service created a network of sentinel stations based in primary care facilities in four Hungarian counties. The aim was to establish a system that will provide valid data on morbidity of selected diseases in Hungary. METHODS: Based on standardized protocols, the participating centres have continuously reported data on the prevalence of cardiovascular diseases, diabetes mellitus, liver cirrhosis, and some malignant diseases, as well as supplying denominator data. The four counties represent both eastern and western parts of Hungary, reflecting the known geographical disparities in health. Each county office enrolled general practitioners maintaining representation in terms of both geography and distribution of settlement size. RESULTS: A total of 73 general practitioners agreed to participate, providing care for 15.6% (138,088 people) of the population in the counties. The population registered with the practices were representative in terms of age and sex of both the participating counties and the entire country. The prevalence of hypertension, diabetes mellitus and liver cirrhosis is high in each county but varies considerably, with higher levels in the western counties, especially among older age groups of both sexes. CONCLUSIONS: The establishment of sentinel stations to collect morbidity data is feasible and sustainable in Hungarian primary care. The data that have been generated provide a valid and comprehensive picture of important aspects of the Hungarian population's health, with important implications for health policy and health service planning. In regions where low prevalence rates of diseases and high mortality rates simultaneously exist special attention is required to explore the background of this caveat. KEY POINTS: Till the end of 1998 no program operated in Hungary engaged with non-communicable disease morbidity data collection, except some hospital-based registries, which failed to produce reliable information. The establishment of sentinel stations to collect morbidity data is feasible and sustainable in Hungarian primary care, the valid morbidity data can be built into the decision making process in health service planning. Regular training, quality control and feedback are important contributors to the success of the program. The prevalence of hypertension, diabetes mellitus and liver cirrhosis is high in each county but varies considerably, with higher levels in the western counties, especially among older age groups of both sexes. More research needed to determine the possible contribution of unknown morbidity and health service utilisation to the different prevalence values in the two parts of Hungary. PMID- 15788801 TI - Smoking by French general practitioners: behaviour, attitudes and practice. AB - INTRODUCTION: This paper examines smoking prevalence, sociodemographic factors and the medical practice of French general practitioners. METHOD: Data from the 1998 cross-sectional national survey of 2,073 GPs. The questionnaire was administered by telephone. A response rate of 67% was attained. Instrumentation included questions about medical practice, sociodemographic characteristics, and health behaviour. Bivariate and multiple logistic regression (MLR) analyses were conducted. RESULTS: Almost one-third (32.1%) of physicians were current smokers. A significantly higher proportion of male (33.9%) were smokers compared to women (25.4%, p<0.001) and men were more likely to be former smokers (49.1% versus 31.7%). Two-thirds of physicians reported recommending nicotine replacement therapy to their patients. MLR shown that former smokers were more likely (OR = 1.51, 95% CI, 1.24-1.83) to indicate that their help in getting patients to quit was not effective compared to smokers. Also, physicians who were 'dissatisfied' with the profession were more likely (OR = 0.75, 95% CI, 0.60-0.92) to report their help as not effective than those who were 'satisfied'. CONCLUSION: These data support the need for greater professional participation in reducing smoking among general practitioners in France and greater education concerning the vital role of physicians in promoting cessation among the general population. KEY POINTS: This study examines smoking habits among French GP's, intervention practices, and opinions about their ability to help patients quit smoking. This study found a smoking prevalence rate of 32.1%, 33.9% of male physicians were smokers and 25.4% of women. This study found a smoking prevalence rate of 32.1%, 33.9% of male physicians were smokers and 25.4% of women. PMID- 15788802 TI - Predictors of perceiving smoking cessation counselling as a midwife's role: a survey of Dutch midwives. AB - BACKGROUND: Smoking during pregnancy can have many serious consequences. As the usual providers of pregnancy care in the Netherlands, midwives could serve as effective counsellors to pregnant women about cigarette smoking. The aim of the present study was to identify relevant factors that hamper or promote the provision of effective smoking cessation advice and counselling. METHODS: Questionnaires were mailed to midwives; 237 (64.4%) were returned. Questions were asked about advantages and disadvantages of giving smoking cessation advice, perceived health benefits for mother and child, smoking behaviour and normative beliefs of colleagues, self-efficacy and role definition of midwives with regard to giving smoking cessation advice. RESULTS: Midwives who have a more positive role definition regarding giving smoking cessation advice are more convinced of the advantages of giving advice, the advantages of quitting for their clients and perceive more support from their colleagues with regard to giving advice. CONCLUSION: In general, midwives were motivated to provide their clients with smoking cessation advice. They were less comfortable with guiding women through the cessation process. Therefore, effective materials and training should be developed to facilitate and stimulate midwives in their role as effective counsellors. PMID- 15788803 TI - Smoking attributable mortality in the community of Madrid: 1992-1998. AB - BACKGROUND: The proportion of smokers and the number of cigarettes consumed per person-year in Madrid is above the European average. To evaluate the impact of smoking in public health smoking attributable mortality was estimated for 1998 and for changes since 1992. METHODS: The number of smoking attributable deaths and years of potential life lost attributable to cigarette smoking for 1992 and 1998 by gender and age group were estimated, based on the population attributable fraction. The relative risks of the Cancer Prevention Study II were used. To compare the two periods of study, a Poisson regression analysis adjusted by age was applied. RESULTS: In 1998, 15.9% of total mortality in the population older than 34 years was attributable to smoking. Lung cancer and chronic obstructive pulmonary disease are the main causes of death in both genders. In the period studied, 1992-1998, the adjusted rates diminished in men in both age groups, while in women they increased in the age group 35 to 64 years. The mortality from lung cancer remained stable in men, increasing in women by 12%. The years of potential life lost decreased by 14% in men and increased by 42% in women. CONCLUSIONS: One in four deaths in men and one in 36 deaths in women are attributed to cigarette smoking. The smoking attributable mortality in males has tended to stabilize, while in women premature mortality is increasing. KEY POINTS: Smoking attributable mortality was estimated for 1998 and for changes since 1992 to evaluate the impact of smoking in the population older than 34 years. MAIN RESULTS: In the period studied, 1992-1998, the adjusted mortality rates attributable to smoking diminished in men, while in women they increased in the age group 35 to 64 years. The main causes of Smoking attributable mortality are Lung cancer and chronic obstructive pulmonary disease in both genders. The mortality from lung cancer remained stable in men, increasing in women by 12%. IMPLICATIONS FOR PUBLIC HEALTH: In the Community of Madrid, health promotion programmes to reduce cigarette smoking need to be intensified, specially in women. PMID- 15788804 TI - Childhood adversities and health variations among middle-aged men: a retrospective lifecourse study. AB - BACKGROUND: Using a lifecourse approach, this study examines whether childhood adversities act on adult health as latent or pathway effects, and whether not only childhood ill health and material deprivation, but also an adverse psychosocial environment in terms of stressful relations with parents contribute to later ill health. METHODS: Lifecourse interviews with 380 men born in 1946 were conducted. Outcome variables were perceived health, number of medical conditions, and activity limitations. Ordinal scales indicating levels of exposures as regards childhood health/growth, childhood material deprivation, stressful relations with parents, educational level, and unhealthy adult behaviours were made. Statistical analyses were performed by non-parametric correlation, logistic regression and OLS regression with dummy variables. RESULTS: Simple regression analyses showed a consistent pattern of more negative health outcomes with higher exposures on each of the lifecourse health determinants, but associations were relatively often not statistically significant. In multivariate analyses, the overall pattern remained with few alterations. The lifecourse determinants differed somewhat in their effects on the three health outcomes. Stressful relations with parents were significantly associated with perceived health and activity limitations. CONCLUSIONS: Childhood adversities influence later health to a large extent as latent effects. Stressful relations with parents were relatively important for two of the health outcomes, suggesting that not only ill health and material deprivation during childhood, but also an adverse psychosocial environment contributes to ill health among middle-aged men. KEY POINTS: The study asks whether childhood adversities influence middle-aged men's health through latent or pathway effects. Adjusted for adult circumstances, childhood health problems were associated with activity limitations and medical problems among men aged 55. Stressful relations to parents during childhood had direct negative effects on perceived overall health and activity limitations. Results indicate that childhood adversities often affect adult health as latent effects. Health promotion policies should consider that childhood psychosocial difficulties may have longterm negative health consequences. PMID- 15788805 TI - Monitoring health inequalities through general practice: the Second Dutch National Survey of General Practice. AB - BACKGROUND: For the second time a plan to monitor public health and health inequalities in the Netherlands through general practice was put into action: the Second National Survey of General Practice (DNSGP-2, 2001). The first aim of this paper is to describe the general design of DNSGP-2. Secondly, to describe self assessed health inequalities in the Netherlands. Thirdly, to present differences in prevalence of chronic conditions by educational attainment using both self assessed health and medical records of GPs. Finally, inequalities in 1987 (DNSGP 1) and 2001 will be compared. METHODS: Data were collected from 96 (1987) and 104 (2001) general practices. The data include background information on patients collected via a census, approximately 12,000 health interview surveys per time point and more than one million recorded contacts of patients with their GPs in both years. The method of statistical analysis is logistic regression. RESULTS: The analyses shows that the lower educated have significantly higher odds of feeling unhealthy and having chronic conditions in 2001. Diabetes and myocardial infarction (GP data) showed the largest difference in prevalence between educational groups (OR 2.5 and 2.4, self-reported data). The way the data is collected (self-assessment versus GP registration) hardly affects the magnitude of the educational differences in the prevalence of chronic conditions. The pattern of health inequalities across chronic conditions in 1987 and 2001 hardly differs. Diabetes doubled in prevalence and health inequalities were not significant in 1987, but compared to the other conditions were largest in 2001 (OR 1.1 versus 2.5). CONCLUSION: Health inequalities were shown to be substantial in 2001 and persistent over time. Socio-economic differences were shown to be similar using self-assessed health data and GP data. Hence, a person's educational attainment did not appear to play a part in presenting health problems to the GP. PMID- 15788806 TI - What information do breast cancer screening programmes provide to Italian women? AB - BACKGROUND: The necessity for building transparent communications on screening, both on risks and benefits, is shared by different sides. There is a general agreement that women cannot express informed participation in a screening programme unless they are given sufficient and adequate information. In the screening context, invitation letters and leaflets often represent the principal source of information. METHODS: The invitation letters and leaflets used by 60 Italian breast cancer screening programmes were collected and evaluated through a score sheet developed to verify what kind of information is provided to women. RESULTS: Fifty-three programmes (88.3%) answered and 47 (78.3%) were included in the analysis because of completeness of the material. Nearly all the programmes provide satisfactory practical information and explanations about the test and the screening aims. Few programmes mention the possibility of some discomfort during the exam (34.0%), quality assessment and operator training (10.6%), double reading (6.4%), radiation risk (6.4%) and data confidentiality (6.4%). 68.1% provide information about recall but none describes what a further assessment involves. Epidemiological and numerical information are present only occasionally. CONCLUSION: Although satisfactorily disclosing some practical information, Italian invitation letters and leaflets remain inadequate in managing side effects and risks. If accurate information has the potential to enable women to make an informed choice, the information inviting them to perform screening test must be improved. Further researcher is needed to evaluate different decision aids to meet women's desires for balanced information. KEY POINTS: Women cannot express informed participation in a breast screening programme unless they are given balanced information both on benefits and adverse effects. Current information screening tools often omit relevent data, fail to give information about pros and cons and ignore uncertainties. Despite these considerations, how much information should be given and how this should be framed remains still to be defined. Further evalutions of different ways of presenting information and women's information needs are urgently required. PMID- 15788807 TI - Screening for breast and cervical cancer in a large German city: participation, motivation and knowledge of risk factors. AB - BACKGROUND: Few data exist on attendance for cervical and breast cancer screening, breast self-examination and knowledge about risk factors for cervical cancer among German women. A population-based survey was performed in the city of Bielefeld in Germany. METHOD: A questionnaire was mailed to 1500 randomly selected women age 25 to 75. Of those, 540 questionnaires were returned and 532 were analysed. RESULTS: Women participating in the survey considered themselves well informed about possibilities for early detection of breast cancer (84.0%). Most information was received from office-based gynaecologists (82.4%). 82.8% had a breast examination by a medical doctor annually and 43.1% practised breast self examination every month. 55.5% of the women had had a mammography, 72.5% gave screening as a reason for a mammogram. Age at first mammography was associated with social class (p<0.001). Cytological smears for early detection of cervical cancer were common and obtained frequently. Age at first Pap smear was associated with social class (p<0.001). 69.9% of the women considered themselves insufficiently informed on risk factors for cervical cancer. Women were poorly informed about risk factors for cervical cancer. Only 3.2% knew that infection with human papillomavirus (HPV) is a risk factor for cervical cancer. Giving a correct answer was associated with social class (p<0.001) but not with age. CONCLUSION: Rates of opportunistic mammography screening were high in the study population. Information on risk factors for cervical cancer was scarce. Efforts should be made to improve women's knowledge about risk factors for cervical cancer. KEY POINTS: What do German women know about screening for breast and cervical cancer offered annually by health insurances at no cost? Most women participating in the survey considered themselves well informed about possibilities for early detection of breast cancer. Over two third of the women considered themselves insufficiently informed on risk factors for cervical cancer. Only 3.2% of the women knew that infection with human papillomavirus (HPV) is a risk factor for cervical cancer. There is a pressing need in Germany for better education of the general public on risk factors for cervical cancer. PMID- 15788808 TI - Perceptions of sexual harassment in Swedish high schools: experiences and school environment problems. AB - BACKGROUND: Sexual harassment in schools is recognized as a public-health problem detrimental to girls' psychosomatic health. This study examines the magnitude of sexual harassment and types of behaviours related to sexual harassment that female students are exposed to in a school environment, and their perceptions of them as problems in school. METHOD: A random sample of 540 female high school students, from all over Sweden, responded to an anonymous self-report mail questionnaire consisting of items related to personal experiences of different behaviours related to sexual harassment during the previous school year. RESULTS: Sexual harassment was identified by 49% of the female students as a problem present in their schools. The most common types were verbal behaviours, such as: sexualized conversations, attractiveness rating, demeaning comments about gender, name-calling, and sexual personal comments. The most common non-verbal displays were: sexualized contact seeking and sexual looks. Behaviours in the sexual assault and teacher-to-student categories were less prevalent. In all four categories, the respondents who reported exposure to a particular behaviour were significantly more likely to identify that behaviour as a problem in their school. However, many non-exposed respondents also perceived such behaviours as problems in their school. CONCLUSIONS: Female high-school students in Sweden are exposed to a variety of inappropriate and/or unacceptable behaviours of a sexual nature, or based on sex, that may infringe their right to a supportive, respectful and safe learning environment or their dignity. Greater efforts are needed to analyse and prevent sexual harassment in schools. PMID- 15788809 TI - Female genital mutilation in the context of migration: experience of African women with the Swiss health care system. AB - BACKGROUND: In contrast to other European countries, the Swiss health care system does not deal with the specific needs of women with female genital mutilation (FGM) in a consistent manner, though migrants from countries where FGM is commonly practised are living in Switzerland. METHODS: For this study, the interaction of women who had undergone FGM, with the Swiss health services was analysed both from the perspective of the women concerned and that of health care professionals (doctors and midwives). The methods used were mainly qualitative, including focus group discussions with 29 women from Somalia and Eritrea and telephone interviews with 37 health care professionals. RESULTS: It is estimated that some 6000 girls and women with FGM could be living in Switzerland. The fact that health care providers rarely see such patients and the absence of professional guidelines give rise to misunderstandings. Counselling of the women is often inadequate, and there is a striking lack of communication about FGM between health care providers and users, as well as within the women's communities. As a surprising finding, reinfibulation is carried out in Switzerland when requested by the patient. CONCLUSIONS: This study shows that at present the specific gynaecological and obstetric health care needs of migrant women who have undergone FGM are not adequately addressed in the Swiss health system. The situation could be easily improved by taking into consideration the fairly simple measures suggested by the women and the health care providers taking part in this study. KEY POINTS: Due to international migration, female genital mutilation (FGM) has become an issue of increasing concern in host countries such as Switzerland. Objectives of this study were to analyze how immigrant women with FGM experience gynaecological/obstetrical care in the Swiss health care system, and to investigate if gynaecologists/obstetricians and midwives treat and counsel FGM related complications adequately. Specific health care needs of women with FGM should--and easily could--be better considered by the Swiss health care system. Recommended measures include capacity building and professional guidelines on FGM for health care providers; and access to culturally appropriate information and self-help groups for the communities concerned. PMID- 15788810 TI - The Finnmark Intervention Study: do community-based intervention programmes threaten self-rated health and well-being? Experiences from Batsfjord, a fishing village in North Norway. AB - AIM: Examine negative side-effects on self-rated health and well-being of a community-based intervention in a fishing community in the Norwegian Arctic. METHOD: A cohort study with quasi-experimental design with one intervention community and three control communities from the same area. There was a baseline screening (1987), three years intervention (1988-1991) and a rescreening (1993). Of the invited (aged 20-62 years in 1987), 668 (64%) males and 656 (72%) females met at both screenings. The main outcome variables were self-rated health, mental health variables, and preoccupation with health. Those answering the relevant questions at both screenings were analysed. High-risk and low-risk males were analysed separately. There were too few females in the high-risk group to do separate analyses. RESULTS: The main findings among the low-risk group were: Males in Batsfjord did not differ from males in the control communities. Among females, there was a reduction in the proportion depressed of 20.4% points in Batsfjord and 9.6% in the control communities (p adjusted=0.060). There was also a decrease in the proportion lonely of 19.2% points and 5.8% respectively (p adjusted=0.005). In the male high-risk group there was a decrease in the proportion lonely by 14.7% points in Batsfjord and an increase by 5.9% in the control communities (p adjusted=0.004). CONCLUSION: We have not found that an intervention programme using local empowerment, mass strategies, and intervention on high-risk cases has serious negative side effects on self-rated health, well being or preoccupation with health. PMID- 15788811 TI - Regional collaboration in public health training and research among countries of South Eastern Europe. AB - It is argued that each country of South Eastern Europe should have its own school of public health. However, a basic prerequisite of modern public health training is the comprehensiveness of the programme and a worldview approach. Most of the countries of South Eastern Europe face the same difficulties to adapt their inherited communist structures of public health training to Western standards. A regional collaboration would facilitate the process of establishing schools of public health in all countries of the region and support the training of public health professionals at all levels. KEY POINTS: South East Europe includes Albania, Bosnia and Herzegovina, Bulgaria, Croatia, Greece, Macedonia, Moldavia, Romania, Serbia and Montenegro, and Slovenia. Public health institutions in South East Europe face similar difficulties to adapt their inherited teaching structures to Western standards. Public health institutions in South East Europe should make a joint effort towards establishing regional training programmes. A regional approach in public health training would enable an efficient use of resources in countries of South East Europe. PMID- 15788812 TI - Preparedness and response to terrorism: a framework for public health action. AB - Political group violence in the form of terrorist actions has become a reality worldwide, affecting the health and economies of populations. As a consequence, preparedness and response are becoming an integral part of public health action. Risk appraisal, preservation of human and civil rights and communications within and between countries are all issues to be considered in the process. The combination of the natural history of terrorist actions and the epidemiological triangle model has been adapted in this paper and suggested as a comprehensive approach for preparedness and action. It covers preparedness (pre-event), response (event) and the consequences (post-event) of a terrorist attack. It takes into account the human factor, vectors and environment involved in each one of the phases. KEY POINTS: Terrorism is a global reality with varying underlying causes, manifestations and impact on the health of the public. Preparedness, response and rehabilitation are an integral part of public health action. Consideration of the pre-event, event and post-event phases in terrorist actions, together with the human factor, vector/agent and environment in each of these phases, offers a framework for public health preparedness, response and rehabilitation. Planning should consider risk assessment, risk communication, inter-sectorial cooperation, enactment of laws and regulations which consider protection of the public's health and civil liberties. Allocation of resources would need to make allowance for maintenance and development of ongoing public health activities. PMID- 15788813 TI - Response to Giacaman: terror toll before Jenin. PMID- 15788817 TI - Prehospital and retrieval medicine. PMID- 15788818 TI - Prehospital care section meeting the readers needs. PMID- 15788819 TI - Early management of adults with an uncomplicated first generalised seizure. AB - A literature review of first seizures in adults was performed and a management algorithm was constructed. This review highlights the importance of a thorough history and examination, routine biochemistry and haematology, an electrocardiogram, selected neuroimaging, discharge planning with driving and lifestyle advice, and follow-up in a specialist clinic. PMID- 15788820 TI - Emergency physicians' practices and attitudes regarding procedural anaesthesia for nasogastric tube insertion. AB - OBJECTIVE: To determine practice and attitudes of emergency physicians regarding procedural anaesthesia for nasogastric tube insertion (NGT). METHODS: Survey of resident/attending emergency physicians working in a tertiary care medical centre. RESULTS: Of 68 physicians, 46 responded: 98% believed that awake and alert patients find NGT insertion uncomfortable/painful; 93% used measures to reduce this, most commonly lubricant gel, topical anaesthetic spray, lidocaine gel, and distraction/use of a child life worker; 28% believed these provided adequate pain control and 37% believed they were inadequate. Topical anaesthetic spray, lidocaine gel, and nebulised/atomised anaesthetics were believed the most practical to administer and 44% actually used these. Nebulised/atomised anaesthetics, systemic anxiolytics, and topical anaesthetic spray were believed the most effective at pain control but only 24% actually used these. While 39% of respondents were satisfied with their current practice, 46% were dissatisfied: 91% would change their practice if new literature were to show a convenient way to effectively reduce this pain. CONCLUSIONS: Emergency physicians do not actually use the measures they believe are most practical/most effective at reducing the pain associated with NGT insertion. Thus, there may be a barrier to the use of these measures. Improvement in procedural anaesthesia for NGT insertion in emergency departments is needed and desired by emergency physicians. PMID- 15788821 TI - Attendance at a hospital emergency department by drivers involved in automobile accidents in Italy. AB - OBJECTIVE: This study investigated the profile of drivers involved in automobile accidents attending a hospital emergency department (ED) in Catanzaro (Italy). METHODS: Car drivers involved in automobile accidents who were registered for emergency care between May 2003 and February 2004 were included in the study. Demographics and details of the accident were collected immediately after admittance, before examination by the medical staff. For each patient, the medical staff completed a form including diagnostic investigations and medical/surgical examination in the ED. RESULTS: Of a total of 424 drivers included in the study 27.4% had conditions that were definitely non-urgent problems. Multiple logistic regression analysis indicated that the use of the ED as a source of non-urgent care was significantly higher among patients who were driving at a lower speed when the accident occurred, among those who presented to the ED before the implementation of the new Italian traffic code, and among those who underwent fewer diagnostic investigations and medical/surgical examinations in the ED. Most of the automobile related lesions occurred in the neck (43.9%) followed by multiple body regions (12.5%) and the upper extremities (10.4%). According to the nature of the injury a third were contusions (34%), followed by pain without physical signs and symptoms (28.8%), and dislocation, sprains, and strains (22.9%). CONCLUSIONS: Development of health promotion and education campaigns is required to prevent the use of the ED as a source of non-urgent care by those involved in automobile accidents. PMID- 15788822 TI - The Barrow-in-Furness legionnaires' outbreak: qualitative study of the hospital response and the role of the major incident plan. AB - OBJECTIVE: To document the organisational response of Furness General Hospital to the large outbreak of legionnaire's disease in April 2002 and assess the contribution made by the hospital's major incident plan. METHOD: Qualitative analysis of interview transcripts and written comments from some staff involved in the management of the incident. Documentary analysis of major incident plan and other written materials. RESULTS: The incident posed considerable managerial and clinical problems and this paper describes how they were overcome. In particular, strategies for dealing with supply (of staff, beds, and resources) and managing demand (by liaising with primary care and the public) seem to have been successful. Many functions necessary for managing the incident were poorly dealt with in the plan, especially procedures for handling the news media and liaison with agencies outside the hospital. Lack of explicit guidance appeared not to hinder the organisational response. There may have been an unspoken high level decision to allow staff to draw on their skills and experience in improvising a response to the initial challenge and learning adaptively as the incident unfolded. There was also evidence that staff disregarded existing job and role boundaries and focused instead on tasks, working flexibly to ensure that these tasks were completed. CONCLUSIONS: Protracted major incidents pose particular management challenges and may benefit from an approach different from that set out in typical major incident plans. Staff must be able to act flexibly and responsively. Some form of checklist or toolkit may be preferable to a detailed plan for some types of incident. PMID- 15788823 TI - Does leucocytosis identify bacterial infections in febrile neonates presenting to the emergency department? AB - OBJECTIVE: This study was undertaken to evaluate the discriminatory power of the peripheral white blood cell (WBC) count to identify bacterial infections in a cohort of febrile neonates (or=38 degrees C, underwent lumbar puncture, and had a WBC count obtained were included. They were divided according to microbiological and radiographic findings into four groups: bacterial infections, viral infections, pneumonia, and negative sepsis evaluations. RESULTS: A total of 69 febrile neonates met the inclusion criteria. The number of neonates in each group was as follows: 8 with bacterial infections, 10 with viral infections, 3 with pneumonias, and 48 with negative sepsis evaluations. There was substantial overlap in WBC counts among the groups. The area under the ROC curve was 0.7231 (95% CI 0.5665 to 0.8797). CONCLUSION: In a cohort of febrile neonates evaluated in the emergency department, the WBC count had modest discriminatory power in identifying neonates with bacterial infections and demonstrated substantial overlap among groups. The present data suggest against the use of any WBC count threshold to identify bacterial infections in febrile neonates presenting to the emergency department. PMID- 15788824 TI - Incidence of injuries caused by dogs and cats treated in emergency departments in a major Italian city. AB - OBJECTIVE: This study aimed to estimate the incidence and characteristics of injuries caused by dogs and cats in the population of a major Italian city. METHODS: The clinical records of all patients attending the emergency department (ED) were obtained from the two main hospitals of the city, covering an estimated population of over half a million. A case was defined as a patient admitted for bite or scratch injuries caused by dogs, cats, or other mammals between 1 January 2000 and 31 December 2002. For each case, the information collected included age and sex of the patient, the anatomical site of the injury, and the species of the animal involved. RESULTS: The average yearly incidence of dog and cat bite/scratches was 58.4 and 17.9 cases per 100 000 residents, respectively. Admissions peaked during the summer months. Dogs accounted for 76.9% and cats for 19.7% of cases. Dog injuries were significantly more common in males and younger individuals. Children younger than 9 years had a significantly higher risk of being bitten on the head, face, or neck. Conversely, injuries from cats were significantly more common in females and older people. CONCLUSIONS: Surveillance of injuries caused by dogs and cats could provide useful information for planning and evaluating public health interventions. Collection of data from hospital EDs may be an appropriate, simple, and quick tool for monitoring the phenomenon and evaluating the associated risk factors. PMID- 15788825 TI - FAST on operational military deployment. AB - BACKGROUND AND OBJECTIVE: Handheld ultrasound, because of its light weight, size, rugged design, and relative simplicity of use is ideal for use on operational military deployment. These machines have been used in the diagnosis of a range of traumatic conditions including abdominal, thoracic, and extremity trauma in the hospital environment, yet few data exist on their use during military operations. This paper presents experience of handheld focused assessment with sonography for trauma (FAST) on operational military deployment. METHOD: Over a two month period, handheld FAST was performed by a single surgeon during the circulation phase of the primary survey in trauma patients presenting to the British Military Hospital in Iraq. RESULTS: Fifteen from casualties underwent a FAST examination. Ten were victims of blunt trauma, two had received injuries anti-personnel mines, and three had penetrating injuries from ballistic trauma. There was one positive FAST, confirmed at laparotomy as bleeding from a liver injury. Thirteen scans were negative and remained negative on repeat FAST at 6 hours. One further patient with a negative FAST underwent laparotomy because of transectory, there was no intra-abdominal blood or fluid at surgery. CONCLUSION: Handheld FAST is a valuable technique for investigating abdominal or thoracic bleeding in single or multiple casualty events on operational military deployment. PMID- 15788826 TI - Foreign bodies in the ear: a simple technique for removal analysed in vitro. AB - OBJECTIVE: Foreign bodies in the ear are mainly encountered in children. This can often pose a problem especially in an accident and emergency department where a microscope or expert help is not routinely available. This paper presents a simple, safe, and effective way of ear syringing. The ease and simplicity of the procedure along with the equipment are described. METHOD AND RESULT: The equipment consists of a "disposable" sterile kit, consisting of a 20 ml syringe, saline at body temperature and 14 or 16 gauge cannula (without the needle). An in vitro experiment was conducted to calculate the pressure generated by the water jet on the eardrum. The pressure was well below the pressure required to burst a tympanic membrane, and hence this technique is safe to use. CONCLUSIONS: Ear syringing is an effective and easy way of removing most foreign bodies. A detailed history and an otoscopic examination must precede the procedure. The novel method of syringing described in this paper with the usual safeguards could be a useful adjunct in the management of this common condition. PMID- 15788827 TI - Agreement between an arterial blood gas analyser and a venous blood analyser in the measurement of potassium in patients in cardiac arrest. AB - BACKGROUND AND OBJECTIVE: Rapid measurement of potassium is crucial in the resuscitation of patients in cardiac arrest. This is often done on an arterial blood gas sample taken during resuscitation and analysed in an emergency department based blood gas analyser. No-one has assessed how accurate or reliable this is when compared with the traditional method of sending a venous sample to the laboratory for standard analysis. This study looked at the agreement between potassium measurements in arterial blood gas samples and venous blood samples in patients in cardiac arrest. METHOD: Arterial and venous blood samples were taken at the same time and analysed in the usual way from 50 patients in cardiac arrest. It was found that the mean difference between each pair of arterial and venous potassium measurements was low at 0.106 mmol/l. However, the standard deviation of these differences and subsequently the 95% limits of agreement were wide (-1.182 mmol/l to 1.394 mmol/l)-that is, 95% of differences will lie between these limits. It is felt that these limits are too wide for safe use in clinical practice. CONCLUSION: Based on these results, it is advised that arterial blood gas analysers should be used with caution to measure potassium in patients in cardiac arrest. PMID- 15788828 TI - Best evidence topic report. Glucagon infusion in refractory anaphylactic shock in patients on beta-blockers. AB - A short cut review was carried out to establish whether a glucagon infusion is of benefit in patients with refractory anaphylaxis. 62 papers were found using the reported search, of which two presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 15788830 TI - Best evidence topic report. Topical analgesia for pain reduction in arterial puncture. AB - A short cut review was carried out to establish whether topical local anaesthetic reduces the pain of arterial puncture. 431 papers were found using the reported search, of which two presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 15788831 TI - Best evidence topic report. Antifibrinolytics for the initial management of sub arachnoid haemorrhage. AB - A short cut review was carried out to gather the evidence for and against the use of tranexamic acid to patients who have suffered subarachnoid bleeding. 267 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date, and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of this best paper are tabulated. A clinical bottom line is stated. PMID- 15788832 TI - Best evidence topic report. Anticoagulation post-cardioversion of acute atrial fibrillation in the emergency department. AB - A short cut review was carried out to establish whether anticoagulation is indicated after emergency department cardioversion of a patient with acute onset atrial fibrillation. 243 papers were found using the reported search, of which none presented any evidence to answer the clinical question. It is concluded that there is no evidence available to answer this question. Further research is needed. PMID- 15788833 TI - Best evidence topic report. Anticoagulation before cardioversion of acute atrial fibrillation in the emergency department. AB - A short cut review was carried out to establish whether anticoagulation is indicated prior to emergency department cardioversion of a patient with acute onset atrial fibrillation. 54 papers were found using the reported search, of which none presented any evidence to answer the clinical question. It is concluded that there is no evidence available to answer this question. Further research is needed. PMID- 15788834 TI - SOCRATES 6 (Synopsis of Cochrane Reviews applicable to emergency services). PMID- 15788835 TI - Mental Health- Consent, the law and depression- management in emergency settings. PMID- 15788836 TI - For debate...: a license to practise pre-hospital and retrieval medicine. PMID- 15788837 TI - A license to practise pre-hospital and retrieval medicine. Commentary from RCGP. PMID- 15788838 TI - A license to practise pre-hospital and retrieval medicine. Commentary from the RCA. PMID- 15788839 TI - A license to practise pre-hospital and retrieval medicine. Commentary from BASMeD. PMID- 15788840 TI - A license to practise pre-hospital and retrieval medicine. Commentary from BASIC [corrected]. PMID- 15788841 TI - A license to practise pre-hospital and retrieval medicine. Commentary from BASICS Scotland. PMID- 15788842 TI - A license to practise pre-hospital and retrieval medicine. Commentary from FAEM. PMID- 15788843 TI - A license to practise pre-hospital and retrieval medicine. Commentary from the Royal College of Surgeons of Edinburgh. PMID- 15788844 TI - Transorbital brain injuries. PMID- 15788845 TI - Intermittent left bundle branch block (LBBB) as a clinical manifestation of myocardial contusion after blunt chest trauma. AB - Injury to the heart in blunt chest trauma is dependent on a number of factors. Symptoms are often non-specific, and there is no gold standard test for diagnosis. Injuries to small areas of the myocardium may only be identified at autopsy. We report a 38 year old man who sustained a number of injuries in a road traffic accident, and in whom the single clinical or ECG abnormality was a left bundle branch block (LBBB); he had a myocardial injury rated as grade II. The patient was treated for his injuries and later discharged. As this is a difficult diagnosis, algorithms of blunt chest trauma may save time and money by avoiding misleading diagnosis and unnecessary monitoring and admissions. PMID- 15788846 TI - Hepatic artery aneurysm: an unusual cause for gastrointestinal haemorrhage. AB - Aneurysms of the hepatic artery are rare. The presenting features include abdominal pain, which may be associated with a mass. More acutely, patients present with signs of hypovolaemia secondary to rupture. The patient reported here presented with gastrointestinal haemorrhage of unknown aetiology. A computed tomography scan showed an aneurysm of a visceral artery. Subsequent angiography confirmed the presence of a leak in the hepatic artery. The patient refused surgical intervention and improved with supportive treatment only. PMID- 15788847 TI - An unusual case of a compound depressed skull fracture after an assault with a stiletto heel. AB - Compound depressed skull fractures occur commonly from assault with various sharp and blunt objects. The use of a stiletto heeled shoe as an offensive weapon has not been reported before as a cause of such an injury. However unusual the history of assault, a clear picture of the mechanism of the injury is vital. Patients with scalp lacerations following a direct blow to the head must be evaluated thoroughly. Early investigation and treatment are necessary to reduce the risk of complications, as illustrated by this case. PMID- 15788848 TI - An aortic dissection in a young weightlifter with non-Marfan fibrillinopathy. AB - Aortic dissection (AD) is an uncommon condition that occurs mainly in the older patient population (>40 years). It is rare in younger people and is usually associated with trauma, Marfan's syndrome, or pregnancy. We report a case of a young weightlifter who died from an AD, and upon autopsy, was diagnosed as having non-Marfan's fibrillinopathy. We recommend that AD should be considered in symptomatic patients with any family history of early cardiac deaths, a history suggestive of a connective tissue disorder (that is, multiple joint surgeries) or who practise weightlifting. PMID- 15788849 TI - Acute glaucoma presentations in the elderly. AB - Acute glaucoma classically presents with severe pain, redness, and reduced vision in the affected eye, and severe cases can also have systemic symptoms. We report three cases of acute glaucoma in elderly patients. The diagnosis of acute glaucoma in a patient who presents with sudden onset of a painful, red eye with reduced visual acuity, a hazy cornea, and a fixed, semi-dilated pupil is comparatively straightforward. However, any patient with headache, malaise, or gastrointestinal disturbance, especially with clinical signs of an acute red eye and reduced vision, should alert doctors to the possibility of acute glaucoma. This is especially important in elderly people, who may not volunteer any specific ocular symptoms. PMID- 15788850 TI - Fatal acute disseminated encephalomyelitis following treated snake bite in India. AB - Snake bite is an important cause of mortality and morbidity in India, with an estimated 35,000 to 50,000 fatal bites occurring annually. The neurological consequences of snake bite are predominantly the result of inhibition of neuromuscular transmission. We describe the first documented case of autopsy proven acute disseminated encephalomyelitis following treated snake bite in a young female. PMID- 15788852 TI - Difficult adherent nail bed dressings: an escape route. PMID- 15788851 TI - Mydriasis due to Datura inoxia. AB - Unilateral, dilated unresponsive pupil may be secondary to a wide range of ocular and neurological disorders. "Gardener's pupil" is a pharmacological mydriasis caused by exposure to plants containing alkaloids. We report a case of mydriasis related to Datura inoxia, and detail a number of plants that have toxic effects. This report emphasises the importance of accurate history taking when evaluating fixed dilated pupil. We recommend that labelling of such poisonous plants should detail the local and systemic effects of accidental exposure, rather than merely the fact that they are poisonous. PMID- 15788853 TI - Dizziness and yield of emergency head CT scan: is it cost effective? PMID- 15788854 TI - [Modern approaches to child and adolescent health assessment at population level]. AB - Despite the notion that the future of our society depends on children; their health is neither a key target in national health programmes, nor a key priority for research. Further improvement in child and adolescent health in Poland depends partly on appropriate data and on indicators applicable for planning, monitoring and evaluating purposes. Mortality statistics do not provide adequate description of the burden of disease in this age group and new indicators are required due to on increase in chronic diseases, and mental health problems. The aim of the paper is to discuss the concept of child and adolescent health and to stress the need of reliable data, correctly collected, analysed and interpreted. The paper compares the biomedical and biopsychosocial paradigms concerning children's health and well-being. Main determinants of health as alternative outcome measures, commonly used in health promotion, are also discussed. CHILD (The Child Health Indicators of Life and Development) project indicators are presented and their availability in Poland is described. A short international comparison based on five indicators related to health and health determinants, showed that Poland, a relatively low-income country in EU, is affected by health problems typical for Central and Eastern Europe (physical, such as relatively high mortality, poor oral health, and psychological, such as lower life satisfaction). However, new problems typical for more developed West European countries are expected to increase (like asthma and eating disorders). On the other hand, by comparison to other countries more children in Poland live in full families with supportive parents. In conclusion, there is a great progress in measuring children's health all over the world, although most of the new approaches have not been yet implemented in Poland. There is a special need to develop and implement multidimensional measures with well-established validity and reliability, such as quality of life scales and health profiles. PMID- 15788855 TI - [The meaning of health evaluation and its assessment in forming healthy behaviors of children and young people]. AB - Healthy behaviour of children and adolescents and their determinants are subject of many research studies in modern psychology and health pedagogy. The most frequently mentioned predictors are: a sense of self-efficacy, coping with stress, optimism and health locus of control. The aim of the conducted study was to try and answer whether such variables as value of health and subjective valuation of own health can be predictors of children and adolescents' health behaviours. The article shows the results of the research on 903 students of primary schools, high schools and secondary schools. The following methods were used: the Juczynski Health Behaviours Inventory, the Juczynski Personal Value Scale (LWO) and VAS scale. The results indicated significant influence of the value of health and its valuation on taking up health behaviours of the researched group. The examined children and adolescents who highly value health take up more wellness behaviours. Those who consider their health to be in a very good condition avoid behaviours and places threatening health, are more active physically and take up health practices more often. PMID- 15788856 TI - [Diagnosing mental health of adolescents on the basis of their subjective assessments]. AB - AIM: the aim of the study was to assess the possibility of diagnosing mental health status of adolescents according to their self-reports. METHOD: 13-years old students from randomly chosen Warsaw schools (N=1123, 54% girls), participated in the school survey. The assessment of health status was based on 3 questions from the HRQOL (Health Related Quality of Life) questionnaire. Respondents were asked to assess the number of days in the past month in which (1) their psychological state was not good, (2) they had problems related to their physical health, and (3) their functioning was impaired because of problems with health. Answers were included in the cluster analysis. RESULTS: it permitted to identify a group of adolescents (5% of the sample) who cannot function as their peers because of health (mainly mental health) problems and a group of those who report that they often have psychological problems (9%). This negative subjective validation was consistent with the results of tests commonly used as psychological indicators of mental health: self-esteem (based on scores in Rosenberg's scale), sense of coherence (SOC-13) and psychological distress (GHQ - General Health Questionnaire). CONCLUSIONS: the study results permit to assess the scale of adolescents' needs for psychological support. Adequate service for groups, mentioned above, is necessary because negative self-perception of health and emotional problems in adolescence are risk factors of mental illness in adulthood. PMID- 15788857 TI - [Development and initial psychometric analysis of the generic quality of life questionnaire for Polish children and adolescents (KIDSCREEN-52)]. AB - BACKGROUND: KIDSCREEN was developed as EC project simultaneously in 13 European countries in ten languages and provided generic measure of quality of life (QOL) in children and adolescents aged 8-18 yrs. An instrument was constructed including 52 items grouped in 10 dimensions, as well as 2 shorter versions, both for children and parents as proxy respondents. AIMS: this study describes the development and initial psychometric evaluation of Polish KIDSCREEN-52 instrument. MATERIAL: the questionnaire was tested in a national mail survey conducted on the representative sample of 1718 children and adolescents aged 8-18 yrs. RESULTS: Polish children and adolescents reported the highest QOL scores for social acceptance and moods and emotions, and the lowest for financial resources and school environment. Gender differences are small in children and but increased in adolescents, especially in self-perception. Cronbach's a ranged from 0.79 to 0.91, and was a little higher in girls than in boys, and higher in adolescents than in younger children. As expected, very high differences were observed between mentally healthy and ill children identified by SDQ (Strengths and Difficulties Questionnaire). To assess the relationship with chronic conditions, the Children with Special Health Care Needs Screener (CSHCN) was applied. Children with special health care needs especially reported lower physical well-being in comparison to healthy children, with moderate effect size. There was a moderate but significant correlation between all ten QOL dimensions measured by KIDSCREEN-52 and other QOL measures applied so far at national level (for example Cantril ladder). CONCLUSION: preliminary results suggested that KIDSCREEN questionnaire could be implemented at the national level and included in health surveys to monitor, evaluate and improve health and subjective well being in children and adolescents. The Polish version of KIDSCREEN-52 questionnaire is reliable and valid, however, more psychometric analyses should be carried out, national norms should be estimated and some international comparisons be conducted. PMID- 15788858 TI - [Self-reported health and life satisfaction in school-aged children in Poland and other countries in 2002]. AB - AIM: to present results of the study concerning: self-reported health, frequency of subjective complaints and life satisfaction among adolescents in Poland and 31 other countries. MATERIAL AND METHODS: the survey was carried out in 2001/02, on representative for each country samples of pupils aged 11, 13, 15 years in the frame of HBSC survey (Health Behaviour in School-aged Children: A WHO Collaborative Cross-national Study). The total sample size was 162306, in Poland it was 6383. The international standard questionnaire was used. RESULTS: the majority of adolescents considered themselves as healthy or very healthy and satisfied with their life. These percentages were lower for girls and decreased with age. Every third pupil experienced two or more subjective complaints more than once a week or daily. The highest percentages of adolescents rating their health as fair or poor and unsatisfied with their life were in countries of the former Soviet Union. Multiple health complaints were experienced most often by adolescents in the Mediterranean countries (Greece, Italy, Israel, Spain). In Poland, in comparison with other countries in the survey, adolescents more often rated their health as very good, but many pupils experienced subjective complaints and felt unsatisfied with their life. PMID- 15788859 TI - [Level of perceived social support and quality of life in children and adolescents aged 8-18 years]. AB - THE OBJECTIVE of the study was to determine the influence of perceived social support on health related quality of life (HRQL) in children and adolescents. MATERIAL AND METHODS: Data was obtained through KIDSCREEN study (Screening for and Promotion of Health Related Quality of Life in Children and Adolescents. A European Public Health Perspective) conducted in Poland in 2003, were used. The representative sample comprised 1718 children and adolescents aged 8-18-years. Full version of the KIDSCREEN questionnaire including 52 items grouped into 10 dimensions was used. A short, 3 items Oslo social support scale was used and three levels of social support were defined. Differences between mean values of quality of life scores in three groups of social support were tested using ANOVA method with F-statistics significance and eta square measure of association (h2). RESULTS showed that 46% of Polish children and adolescents reported high, 44% middle and 10% low level of perceived social support. Girls perceived social support better than boys and children better than adolescents. The study showed that the level of perceived social support had essential and positive influence on the quality of life in children and adolescents in all ten dimensions (p<0.001). Children with insufficient social support reported lower quality of life in comparison to well supported, especially in Parents Relations and Home Life dimension, h2=0.186. Difference between boys and girls and three age groups under study was the highest in the low supported group and then leveled out when social support increased. CONCLUSION: Social support, especially provided by parents, increases the quality of life in children and adolescents. PMID- 15788860 TI - [Risk behaviors syndrome and subjective health and life satisfaction in youth aged 15 years]. AB - THE PURPOSE of this study was to define risk behaviour syndrome and its influence on subjective health and life satisfaction in adolescents. MATERIAL AND METHODS: The study was based on the HBSC study (HBSC -- Health Behaviour in School Aged Children. A WHO Cross-National Collaborative Study) carried out in Poland in 2002. The sample (N=2152) was selected as representative of pupils from III grade of secondary schools (mean age 15.7). Multivariate logistic regression models were estimated to assess the risk of poor health, frequent psychological or somatic complaints and low life satisfaction in relation to number of escalated risk behaviours, gender, place of residence, family affluence and school achievements. RESULTS: The index of multiple escalated risk behaviours was derived by summing six components: every day smoking, frequent episodes of drunkenness, drug abuse, early sexual initiation, frequent bullying and fighting (alpha-Cronbach=0.696). Almost 43% of adolescents reported at least one out of six behaviours, while 12%, three or more. The number of risk behaviours is increasing dramatically in adolescents from lower social classes and in students with poor school achievement. Boys are less likely to report poor health and low life satisfaction, but more likely than girls to be involved in multiple health compromising behaviours. Adolescents engaged in 4 or more risk behaviours, comparing to not engaged, were at higher risk of poor health (OR=2.39; CI(OR)=(1.51-3.78)) and low life satisfaction (OR=1.65; CI(OR)=(1.06-2.58)). In girls even single risk behaviour resulted in negative health outcome. CONCLUSIONS: Psychoactive substances abuse, early sexual initiation and frequent contact with violence form internally consistent syndrome of risk behaviours in youth. Adolescents from poor families and with poor school achievements are more likely to be involved in multiple risk behaviours, especially in deprived urban areas. Subjective well-being is highly associated with the expression of risk behaviours. PMID- 15788862 TI - [Social determinants of repeated injuries in pupils aged 11-15 years, in Poland]. AB - AIM: To analyse relationships between selected elements of family, school and peer environments and susceptibility to medically treated repeated injuries in pupils aged 11, 13 and 15 yrs in Poland. MATERIAL AND METHODS: The survey was carried out in 2002 on the representative sample of pupils (N=6383), within the Health Behaviour in School-aged Children: A WHO Collaborative Cross-national Study (HBSC). RESULTS: Outcomes of the study indicate that frequency of repeated injuries amounts to 8.6% and rises for pupils from incomplete families (for children living with stepmother OR=3.6, 95% CI 1.40-8.99), being in touch with many friends (OR=2.35, 95% CI 1.82-3.03), whose parents are not ready to help them with school problems (OR=4.28, 95% CI 2.24-8.18) and teachers are not friendly (OR=2.79, 95% CI 1.78-4.38). CONCLUSIONS: Improving relations in families and schools can help in decreasing the frequency of repeated injuries among adolescents. PMID- 15788861 TI - [Subjective health complaints and psychological distress in adolescents aged 15 19 years in Poland]. AB - AIM: To analyse the relationships between selected subjective health complaints and psychological distress of 15-19 yrs old adolescents in Poland. MATERIAL AND METHODS: The health survey of the population was conducted by the Central Statistical Office in 1996 in 19203 households inhabited by 62749 people. Dates were received from 5407 adolescents of 15-19 yrs, using AZD-2 questionnaire. Questions concerning subjective health complaints and psychological distress, tested by modified scale GHQ-12 (General Health Questionnaire), was answered by 3708 adolescents. RESULTS: Subjective health complaints reported 1.9-27.5% of adolescents in the age of 15-19. They experienced feeling nervous and irritability the most often (more than 20% under survey), the most rarely -- insomnia and apathy. Every tenth questioned adolescent at the age 15-19 experienced psychological distress. Psychological distress occurred more often in girls, people living in town and raised with age. Risk of subjective health complaints was higher for people with distress. The highest risk (8 times) was for apathy and insomnia. Psychological distress increased the risk of subjective health complaints for boys more than for girls. PMID- 15788863 TI - [Unfounded dieting among adolescent girls and socio-economic status of their families]. AB - AIM OF THE STUDY was to examine: the prevalence of dieting and other weight control behaviour among adolescent girls without body mass excess (BMI 90 percentile or less); relationship between weight control practices and the socio economic status of the girls' families by age and place of residence. MATERIAL AND METHODS: Data from HBSC (Health Behaviour in School-aged Children: A WHO Collaborative Cross-national Study) surveys, carried out in Poland in 2002 in representative sample of girls aged 11.7; 13.7 and 15.7 (N= 2774) was used. An international standard questionnaire (anonymous) has been chosen as an instrument for data collection. Two indicators of socio-economic status (SES) were developed by researchers in the HBSC study: FAS (Family Affluence Scale) and PFW (Perceived Family Wealth). RESULTS: 18.8% of adolescent girls were engaged in dieting and other weight control behaviour, 25.1% were in the risk group (girls who think that they should loose some weight). The prevalence of these behaviours increase with age, is higher among girls living in cities than in villages. The prevalence of weight control behaviour is higher among girls from families with higher SES. The risk of unfounded weight control behaviour is higher among 15-year old, girls living in town in families with higher SES. CONCLUSIONS: Unfounded dieting and other weight control practices among adolescent girls should be recognized as a new risk behaviour; future studies are necessary for assessment of causes and range of these behaviours as the basis for development of preventive programmes. PMID- 15788864 TI - [The process of disclosure of an HIV/AIDS problem in the family to children--an exploratory study]. AB - AIM: The aim of the study was to describe the process of disclosing HIV/AIDS problem in the family to children and to explore its possible determinants. MATERIAL AND METHODS: The investigations were carried in 37 children, aged 4-16 years, living in families with HIV/AIDS problem (25 children were HIV+ and 12 HIV ) and in their parents/ caregivers. Empirical material consisted of protocols of clinical interviews with parents and children, observation of child's behaviour in different contexts and results of selected projective tests. RESULTS: Only six children (3 were HIV+ and 3 HIV-) had been informed about the problem of HIV/AIDS by the parents and could freely talk with them on that subject. The most important reasons impeding the process of disclosure according to the parents were: risk of depriving the child of a happy childhood and fear that the child may not be able to keep the family secret. As a result of cluster analysis four distinct subgroups of the children under study were discerned. The largest number of emotional and behavioural problems was noted in the case of older, infected children, who were quite aware of their serious medical condition and whose parents were in the phase of secrecy or exploration in the process of disclosure. CONCLUSIONS: The process of disclosing HIV/AIDS problem in the family by parents to children requires taking into account many different factors. Providing professional psychological services may be of great importance in many instances. PMID- 15788868 TI - Tendon proteoglycans: biochemistry and function. AB - Tendon remodeling occurs in response to changes in loading and mobilization. Though the normal mechanical function depends on precise alignment of collagen fibrils, it is proteoglycans that regulate collagen fibrillogenesis and thus, indirectly, tendon function. In this paper we discuss the basic biochemical structure of several members of two proteoglycans families. Decorin, biglycan, fibromodulin and lumican, all members of the small leucine-rich proteoglycans family, bind to collagen fibrils and are active participants in fibrillogenesis. Aggrecan and versican, two members of large modular proteoglycans or lecticans, and their partner hyaluronan likely provide tendon tissues with a high capacity to resist high compressive and tensile forces associated with loading and mobilization. We present data from our laboratory showing that proteoglycans and glycosaminoglycan content increases not only with growth but also with loading of young avian gastrocnemius tendons. Specifically, an increase in the content of keratan sulfate, chondroitin sulfate and hyaluronan was observed. Moderate exercise for several weeks led not only to a further increase in total proteoglycans content but also to qualitative changes in proteoglycan make up. PMID- 15788867 TI - Development of tendon structure and function: regulation of collagen fibrillogenesis. AB - In the tendon, the development of mature mechanical properties is dependent on the assembly of a tendon-specific extracellular matrix. This matrix is synthesized by the tendon fibroblasts and composed of collagen fibrils organized as fibers, as well as fibril-associated collagenous and non-collagenous proteins. All of these components are integrated, during development and growth, to form a functional tissue. During tendon development, collagen fibrillogenesis and matrix assembly progress through multiple steps where each step is regulated independently, culminating in a structurally and functionally mature tissue. Collagen fibrillogenesis occurs in a series of extracellular compartments where fibril intermediates are assembled and mature fibrils grow through a process of post-depositional fusion of the intermediates. Linear and lateral fibril growth occurs after the immature fibril intermediates are incorporated into fibers. The processes are regulated by interactions of extracellular macromolecules with the fibrils. Interactions with quantitatively minor fibrillar collagens, fibril associated collagens and proteoglycans influence different steps in fibrillogenesis and the extracellular microdomains provide a mechanism for the tendon fibroblasts to regulate these extracellular interactions. PMID- 15788870 TI - Metabolic activity and collagen turnover in human tendon in response to physical activity. AB - Connective tissue of the human tendon plays an important role in force transmission. The extracellular matrix turnover of tendon is influenced by physical activity. Blood flow, oxygen demand, and the level of collagen synthesis and matrix metalloproteinases increase with mechanical loading. Gene transcription and especially post-translational modifications of proteins of the extracellular matrix are enhanced following exercise. Conversely, inactivity markedly decreases collagen turnover. Training leads to a chronically increased collagen turnover, and dependent on the type of collagen also to some degree of net collagen synthesis. These changes modify the biomechanical properties of the tissue (for example, viscoelastic characteristics) as well as the structural properties of the in collagen (for example, cross-sectional area). Mechanical loading of human tendon does result in a marked interstitial increase in growth factors that are known potentially to stimulate synthesis of collagen and other extracellular matrix proteins. Taken together, human tendon tissue mounts a vigorous acute and chronic response to mechanical loading in terms of metabolic circulatory changes as well as of extracellular matrix formation. These changes may contribute to training-induced adaptation of biomechanical properties consisting of altered resistance to loading and enhanced tolerance to strenuous exercise. Understanding of such changes is a pre-requisite in the development of measures aimed at prevention of overuse tendon injuries occurring during sport, work or leisure-related activities. PMID- 15788869 TI - Structure, function and adaptation of bone-tendon and bone-ligament complexes. AB - Mechanical and physiological processes contribute to joint tissue adaptations during growth and exercise and after injury. Those adaptations are often in response to the mechanotransductive stimuli linked to the transmission of forces across these load-bearing structures. Muscle-tendon interactions have been explored during skeletal loading and describe the relation of sarcomere shortening at the expense of tendon lengthening(1,2). The effects of load transmission through the bone-tendon and bone-ligament complexes, however, have not been studied as extensively, although both disuse and exercise will alter the stiffness of these significant structures. Recently, however, renewed interest has emerged about the pathogenesis underlying enthesopathies and enthesitis, and investigators are beginning to reveal the intricacies of bone-tendon and bone ligament complexes(3,4,5). Here, we summarize the structure and function of the types of entheses between bone-tendon and bone-ligament, and relate how mechanical loading leads to functional adaptation, and at times, entheseal pathophysiology. PMID- 15788871 TI - In vitro culture decreases the expression of TGF(beta), Hsp47 and type I procollagen and increases the expression of CTGF in avian tendon explants. AB - Weight-bearing tendons in many species, including humans, chickens and horses, are prone to failure, in many cases without a discernible cause. The normal function of the tendon depends on the proper assembly of fibrils of type I collagen, the main structural component of the tendon. We studied the effect of in vitro culture, temperature (37 degrees C vs. 43 degrees C) and wounding on the expression of mRNAs for several collagen regulators, transforming growth factor beta (TGF(beta)), heat shock protein 47 (Hsp47) and connective tissue growth factor (CTGF), in chicken embryonic gastrocnemius tendon explants. The expression of mRNAs for TGF(beta) and Hsp47, a chaperone of collagen assembly, remained strong during the first day of in vitro culture, but then it decreased, slightly more at higher temperature. Additional injury in selected tendons had no significant effect on the levels of TGF(beta) and Hsp47 mRNAs. Likewise, the level of immunostained type I procollagen also decreased with the length of culture. The expression of CTGF gradually increased from 0 at the time of tendon removal with the duration of culture to strong after three days of culture when the expression of TGF(beta) and Hsp47 was low. We conclude that in vitro culture over the period of several days rather than an increase in temperature or additional wounding decreases the expression of TGF(beta), Hsp47 and type I procollagen and increases the expression of CTGF. PMID- 15788872 TI - Histochemistry defines a proteoglycan-rich layer in bovine flexor tendon subjected to bending. AB - Mid-substance fibrocartilage develops in bovine deep flexor tendon at the point where the tendon wraps under sesamoid bones of the foot and receives transverse compressive loading during locomotion. Fibrocartilage extends several millimeters into the tendon at this location and the proteoglycan-rich tissue stains intensely with Alcian blue. Using histochemical techniques we demonstrate the presence of aggrecan, type VI collagen, and hyaluronic acid in the extracellular matrix of this region of tendon. Biglycan staining was localized to the cells, however. Adjacent to the fibrocartilage, at the outer curvature of the tendon as it bends, the tissue resembles typical tensile tendon with dense bundles of linearly arranged collagen. Longitudinal sections revealed discrete layers of Alcian blue-stained material between the collagen bundles. We demonstrate that these layers of loose matrix also contain aggrecan, type VI collagen, and hyaluronic acid. However, the dense collagen bundles of this region are negative for these components. Transverse sections of tendon in the area adjacent to fibrocartilage show a distinct Alcian blue-stained structure surrounding vascular elements at the point where several fiber bundles come together. This is concluded to be the same structure as the Alcian blue-stained layers seen in longitudinal sections. These observations suggest that proteoglycan-rich matrices in tendon subjected to mechanical loading other than pure tension may serve multiple roles. Such matrices can not only provide compressive stiffness and separate and lubricate collagen bundles that move relative to each other, but may also protect the integrity of vasculature in tendon subjected to bending and shear. PMID- 15788873 TI - Early responses to mechanical load in tendon: role for calcium signaling, gap junctions and intercellular communication. AB - Tendon and other connective tissue cells are subjected to diverse mechanical loads during daily activities. Thus, fluid flow, strain, shear and combinations of these stimuli activate mechanotransduction pathways that modulate tissue maintenance, repair and pathology. Early mechanotransduction events include calcium (Ca2+) signaling and intercellular communication. These responses are mediated through multiple mechanisms involving stretch-activated channels, voltage-activated channels such as Ca(v)1, purinoceptors, adrenoceptors, ryanodine receptor-mediated Ca2+ release, gap junctions and connexin hemichannels. Calcium, diacylglycerol, inositol (1,4,5)-trisphosphate, nucleotides and nucleosides play intracellular and/or extracellular signaling roles in these pathways. In addition, responses to mechanical loads in tendon cells vary among species, tendon type, anatomic location, loading conditions and other factors. This review includes a synopsis of the immediate responses to mechanical loading in connective tissue cells, particularly tenocytes. These responses involve Ca2+ signaling, gap junctions and intercellular communication. PMID- 15788874 TI - Evaluating the viscoelastic properties of biological tissues in a new way. AB - In this paper, a new method for evaluating the viscoelastic properties of biological tissues such as tendons and ligaments is presented. This method obtains the complex modulus of these tissues to characterize their viscoelastic properties. With this method, the stresses and strains measured in time are first transformed (using FFT), and the complex modulus is then obtained. The complex modulus contains sufficient information about the viscoelastic characteristics of the biological tissues. With this method, the mechanical properties of biological tissues can be measured without making apriori assumptions regarding their structures, and the measurements can be made in real time. PMID- 15788876 TI - [Indication of drug therapy in patients with chronic hepatitis B]. PMID- 15788877 TI - [Monitoring patients with chronic HBV infection during antiviral therapy]. PMID- 15788878 TI - [Chronic hepatitis B. Dose and treatment duration of regimen]. PMID- 15788879 TI - [Antiviral therapy in hepatitis B carriers undergoing immunosuppressive treatment or cytotoxic chemotherapy]. PMID- 15788880 TI - [Prophylaxis against hepatitis B recurrence following liver transplantation]. PMID- 15788881 TI - [The efficacy of lamivudine in Korean patients with chronic hepatitis B]. PMID- 15788882 TI - [The efficacy of alpha-interferon in Korean patients with chronic hepatitis B]. PMID- 15788883 TI - [HBV-specific CD8+ T cells for sustained HBeAg seroconversion after lamivudine therapy]. AB - BACKGROUND/AIMS: Viral suppression of the hepatitis B virus (HBV) can be induced by lamivudine, but the relapse seen in many patients after cessation of lamivudine therapy is troublesome. We thought that the host immune response is important to prevent viral relapse. We compared the frequency of HBV-specific CD8+ T cells in the peripheral blood and their expansion capacity after exposure to viral antigen between the patients showing sustained HBeAg seroconversion after use of lamivudine and those patients without sustained response. METHODS: We analyzed HBV-specific CD8+ T cells that were isolated from the blood of 14 patients with HLA-A2 who showed lamivudine induced HBeAg seroconversion (HBV DNA < 0.5 pg/mL, and the cells were negative for HBeAg) at the end of lamivudine therapy. The purified T cells were directly stained ex vivo, after they had been stimulate with synthetic peptide, using the HBV core 18-27-specific HLA tetramer (Tc 18-27) and monoclonal antibody to CD8. The HBV viral load was quantified by the Amplicor HBV Monitor assay. RESULTS: In patients with a sustained HBeAg response (the sustained group) for a duration of 15.5 months of follow-up, the median number of Tc 18-27 cells out of the 5 X 10(4) CD8+ T cells was 49.5 (15 135). On the contrary, in patients who experienced relapse (the relapsed group) during a median of 7.5 months of follow-up, the median number of Tc 18-27 cells out of the 5 X 10(4) CD8+ T cells was 13.5 (0-95). Especially, among patients with a viral load of HBV DNA < 1 X 10(3) copies at the end of treatment, the median number of Tc 18-27 cells out of 5 X 10(4) CD8+ T cells was 87 (45-135) in sustained group compared to 12 (6-50) in the relapsed group. All patients in the sustained group demonstrated a vigorous expansion of the core 18-27-specific CD8+ T cells after stimulation with viral peptide, in contrast to only 3 out of 8 patients in the relapsed group. CONCLUSIONS: This study demonstrates that the frequency and functional responsiveness of the circulating HBV-specific CD8+ T cells may be important for obtaining a sustained HBeAg response to lamivudine. PMID- 15788884 TI - [The prevalence and clinical characteristics of hepatitis-delta infection in Korea]. AB - BACKGROUND/AIMS: The prevalence of hepatitis delta virus (HDV) infection has been estimated as being approximately 5% among global HBsAg carriers. The anti-delta positive rate in Koreans had been reported as being 0.85% in 1985. While the prevalence of HBV has been decreased from nearly 10% to 5% during the past twenty years, there have been no more studies on the anti-delta prevalence in Koreans. The aim of this study was to estimate the anti-delta prevalence in Koreans and to study the clinical characteristics of anti-delta positive patients in a single center. METHODS: Serum anti-delta was measured in one hundred ninety four HBsAg positive patients who were admitted to our hospital from February 2003 to August 2003. We checked the genotypes of the HBV in the anti-delta positive patients. The clinical features of the anti-delta positive patients were compared to those clinical features of the anti-delta negative patients from the aspect of age, gender, mode of transmission, the positivity of HBeAg and serum HBV DNA. RESULTS: Serum anti-delta was positive in seven patients among the 194 subjects, giving a 3.6% positive rate. Among these seven patients, six had hepatocellular carcinoma (HCC) and the other one had cholangiocarcinoma. All of the anti-delta positive patients had the C genotype of HBV. The anti-delta positive patients showed significantly suppressed HBV DNA replication compared to the anti-delta negative patients. CONCLUSIONS: In Koreans, anti-delta was positive mainly in HCC patients with an approximate prevalence of 4%, and this rate has not changed much for the past twenty years. HBV DNA replication was suppressed by HDV infection. PMID- 15788885 TI - [Relationship between the severity of liver damage and the serum leptin level for nonalcoholic fatty liver disease]. AB - BACKGROUND/AIMS: Nonalcoholic fatty liver disease (NAFLD) represents a spectrum of conditions that are mainly characterized histologically by macrovesicular hepatic steatosis. There are two histologic patterns of NAFLD: simple steatosis alone and steatohepatitis. The factors leading from simple steatosis to nonalcoholic steatohepatitis (NASH) are still obscure. The datas from several studies have suggested that leptin could be involved in the progression from hepatic steatosis to steatohepatitis including the fibrosis. We evaluated serum leptin levels in patients with NAFLD to determine whether any relationships existed between the leptin levels and the severity of hepatic inflammation or fibrosis. METHODS: We studied 62 patients with NAFLD who were diagnosed at the Hallym University Sacred Heart Hospital from July 2001 to May 2004. We measured the serum leptin level in all cases and liver biopsy samples were obtained from 31 cases. The liver biopsy specimens were graded according to methods described by Brunt. Spearman rank correlations were used to detect the associations between the serum leptin and the various anthropometric and biochemical variables. The relationship between the histologic severity and the serum leptin level was evaluated with logistic regression analysis. RESULTS: Serum leptin levels correlated with insulin, c-peptide, ALT and homeostasis model assessment insulin resistance, but not with BMI, age and gender. Serum leptin level also correlated with hepatic fibrosis, but not with hepatic steatosis or inflammation. However, the serum leptin level was not a significant independent predictor of the grade of hepatic steatosis, inflammation and fibrosis on the univariate analysis. CONCLUSIONS: The serum leptin level was not an independent predictor of the severity of liver damage in NAFLD. PMID- 15788886 TI - [The comparative results of radiofrequency ablation versus surgical resection for the treatment of hepatocellular carcinoma]. AB - BACKGROUND/AIMS: Although surgical resection remains the gold standard of therapy for hepatocellular carcinoma (HCC), only selected patients can undergo resection because of the severity of the underlying cirrhosis or due to the diffuse distribution of the tumor. Radiofrequency ablation (RFA) has recently shown comparable results to surgical resection for the treatment of HCC. We compared the results of RF ablation and surgical resection for the treatment of HCC. METHODS: From January 2000 to December 2002, one hundred-sixty patients who had undergone surgical resection or RFA were analyzed retrospectively. The patients with a tumor size less than 5 cm in diameter, with less than 3 tumors in number, with tumor having a Child-Pugh class A classification and no evidence of extrahepatic metastasis were enrolled in the study. The recurrence pattern was classified into local and distant recurrence. We compared the recurrence patterns, the survival rates, the recurrence rates and the complications between the two groups. RESULTS: 1) The local recurrence rate was 9.8% for surgical resection and 18.2% for RFA and the distant recurrence rate were 32.8% and 28.3%, respectively. 2) The 1-, 2- and 3-year overall cumulative survival rates after RFA and surgery were 95.8%, 86.8%, 80.0%, 98.3%, 87.0% and 77.4%, respectively. 3) The incidence of complication was similar between the two groups. CONCLUSIONS: Radiofrequency ablation shows comparable results to surgical resection for the treatment of HCC. Therefore, RFA should be considered as the treatment of choice those patients who are not candidates for resection. However, intrahepatic recurrence of tumor after RFA was as frequent as that seen after surgical resection. Further investigation is warranted to clarify whether the current RFA technology could offer improved long-term results. PMID- 15788887 TI - Prostaglandin A2 induces caspase-independent apoptosis in hepatocellular carcinoma cells. AB - BACKGROUND/AIMS: Prostaglandin (PG) A2 has been reported to inhibit the growth of hepatocellular carcinoma cells via activation of apoptosis, although the molecular mechanisms involved have not been clarified, yet. To investigate the mechanism of the PGA2-induced apoptosis, we analyzed the activation of caspases during the apoptosis of hepatoma cell lines. METHODS: Induction of apoptosis by PGA2 in hepatoma cell lines, Hep 3B and Hep G2, was assessed by DAPI staining of nuclei and agarose gel electrophoresis of genomic DNA. The involvement of caspases was analyzed by immunoblot analysis of poly ADP-ribose polymerase (PARP) and by checking the effect of caspase inhibitors on PGA2-induced apoptosis. RESULTS: PGA2 inhibited the growth of Hep 3B and Hep G2 cells, accompanying nuclear condensation and fragmentation, and genomic DNA laddering, which are the hallmarks of apoptosis. The PARP was not cleaved during the apoptosis of Hep 3B and Hep G2 cells and caspase inhibitors such as z-VAD-Fmk and z-DEVD-Fmk exerted no effect on the PGA2-induced apoptosis. CONCLUSIONS: These results suggest that PGA2 induces apoptosis in Hep 3B and Hep G2 cells via caspase-independent pathway. PMID- 15788888 TI - [A case of vanishing bile duct syndrome associated with hypersensitivity to allopurinol]. AB - Allopurinol is frequently used for the treatment of hyperuricemia and gout. Sometimes, a life-threatening reaction develops, as is illustrated by the following case report. We describe a 60-year-old male patient who was treated with allopurinol because of asymptomatic hyperuricemia, and he was presented with fever, skin rash, eosinophilia, worsening renal function and vanishing bile duct syndrome. In this report, we discussed vanishing bile duct syndrome as a serious side effect of allopurinol, and we briefly reviewed the etiology, prevention, and treatment modalities for vanishing bile duct syndrome. PMID- 15788889 TI - [Primary adenosquamous carcinoma of the liver: a case report]. AB - Primary adenosquamous cell carcinoma of the liver, a variant of cholangiocarcinoma is very rare disease and its prognosis is known to be very poor. We describe here a case of primary adenosquamous carcinoma of the liver. The patient was a 60-year-old woman who complained of an eight-month history of intermittent upper abdominal pain. Abdominal computed tomography scan revealed a low density mass and intrahepatic duct stones within the dilated left intrahepatic duct in the left lateral segment. Needle biopsy of the mass was done and the histologic diagnosis was cholangiocarcinoma. Left lobectomy was then performed. Upon gross examination, there was a well defined solid mass and intrahepatic duct stone in the dilated bile duct. Microscopically, the tumor was composed of both adenocarcinoma and squamous cell carcinoma. PMID- 15788890 TI - [Inflammatory pseudotumor of the liver]. PMID- 15788891 TI - [Completely ablated hepatocellular carcinoma by percutaneous radiofrequency thermal ablation]. PMID- 15788892 TI - The cultural and philosophical aspects of pressure, massage, and touch healing as alternative therapies. AB - Integrative medicine is becoming more popular, not only in developing nations, but also in countries such as the United States. Acupuncture, which is practiced more than any other kind of alternative medicine, is based on the philosophy and culture of old Chinese healing. Pressure, massage, and touch healing depend on stimulation by the fingers rather than needles, although they are based on acupuncture principles. Touch has social, educational, cultural, and humanitarian aspects. Touch therapy has been shown to have excellent effects on children's growth, development, and emotional well-being. In adults, a 15-minute rubdown by fingertips can lead to relaxation and sleep. It is also useful for premature babies. Such therapies were practiced in the Old World, but have been rediscovered in the New World. They represent some aspects of the old, noble art of healing. PMID- 15788893 TI - Oral contraceptives: their mode of action and dermatologic applications. PMID- 15788894 TI - Diagnosis and treatment of basal cell nevus syndrome. PMID- 15788895 TI - Pigmentary mosaicism of hyperpigmented type in two sisters. AB - A 17-year-old girl and her 15-year-old sister presented with progressively increasing streaks of reticulate hyperpigmented macules arranged in a whorled pattern over the trunk and extremities (Figure), which appeared soon after birth. The face, palms, soles, eyes, and mucous membranes were spared. Both parents were unrelated Saudis. There was no history of any preceding eruption or any associated systemic abnormality, except for recurrent oral ulcers in both patients for several years. The younger one also had one episode of genital ulcers. Dermatological examination of both parents and the patients' three brothers and one sister were normal with no history of oral ulceration. Examination revealed few aphthous ulcers in either cases, but no joint or eye symptoms. Pathergy testing in both cases was negative. Ophthalmological examination and consultation revealed no signs of Behcet's disease. Routine blood tests did not show any abnormalities. Skin biopsies were taken from pigmented and normal skin in both patients. Histopathological examination of pigmented skin in both cases revealed basal cell hyperpigmentation with pigmentary incontinence. Similar features but in milder form were seen in the biopsies of normal skin in both cases. PMID- 15788896 TI - Ribozyme cleavage leads to decreased expression of fibroblast growth factor receptor 3 in human multiple myeloma cells, which is associated with apoptosis and downregulation of vascular endothelial growth factor. AB - The aim of this study was to investigate the fibroblast growth factor receptor 3 (FGFR3) mRNA cleavage by ribozymes targeting FGFR3, effect of growth inhibition and associated with mechanism on multiple myeloma (MM). We designated two ribozyme-expressing plasmids that target the FGFR3 genes, Rz52 and Rz32. In vitro catalytic activity of Rz52 and Rz32 in KMS11 cells decreased FGFR3 mRNA expression to 45% (p < 0.05) and 80% (p < 0.5), respectively, of that of the control. In vivo examination of the Rz52-transfected KMS11 clone showed that FGFR3 mRNA expression decreased to 20% (p < 0.05) of the control. In the Rz52 transfected H929 clone, FGFR3 mRNA decreased to 50% of the control. Protein expression of FGFR3 decreased to 70% of the parental KMS11 and H929 clones. DNA synthesis in the Rz52-transfected KMS11 clone decreased to 20% of that of the control, whereas the viability of cells decreased to 2% (p < 0.01) of that of the control. Ribozyme cleavage-associated increase in apoptosis of Rz52 KMS11 transfectants was twice that of the control. The inhibition of FGFR3 expression by ribozymes was associated with decreased vascular endothelial growth factor (VEGF) expression and upregulation of Flt-1 but not of the KDR receptor. Our data indicate that FGFR3 is an important cell survival and antiapoptotic factor for MM cells and that ribozyme-targeted downregulation of FGFR3 might be useful as a novel therapeutic intervention in MM characterized by t(4;14). PMID- 15788897 TI - Transfection protocol for antisense oligonucleotides affects uniformity of transfection in cell culture and efficiency of mRNA target reduction. AB - In an effort to optimize the transfection of cell lines with antisense oligonucleotides, we examined cellular accumulation of a labeled oligonucleotide by flow cytometry. We were surprised to observe that a routinely used transfection protocol, a fixed lipid/oligonucleotide ratio, resulted in variable transfection efficiency depending on the concentration of oligonucleotide used. A significant population of cells, especially at lower doses of oligonucleotide and cationic lipid, were untransfected. We investigated lipid/oligonucleotide ratios, different lipid preparations, and different cell types and found that these variables did not alter the percentage of cells transfected at these lower doses of oligonucleotide. However, when lipid-oligonucleotide complexes were formed at the high dose and then diluted into a solution of lipid or a complex of lipid and unlabeled, negative control oligonucleotide, a constant percentage of cells was transfected. Under these conditions, mRNA target reduction dose-response curves were also shifted to lower doses. We hypothesize that poor transfection observed at a low concentration of lipid-oligonucleotide complex when diluted in medium is due to loss of active complexes, either by adsorption to the substrate or by changes in physical characteristics of complexes. By maintaining a constant lipid concentration, more consistent transfection was achieved. PMID- 15788898 TI - Silencing of human c-myc oncogene expression by poly-DNP-RNA. AB - Deregulation of c-myc oncogene expression drives the progression of many different types of cancer. Recent experimental data suggest that even brief inhibition of c-myc expression may be sufficient to permanently stop tumor growth and induce regression of tumors. Previous efforts in developing an inhibitor to silence the c-myc gene were hampered by low efficacy and lack of sequence specificity. Here, we report the synthesis of an antisense RNA inhibitor based on a new 21-nt sequence on a poly- DNP-RNA platform that can specifically inhibit cancer cell growth by silencing c-myc gene expression. Both c-myc mRNA and protein levels were significantly decreased in MCF-7 cells following treatment with this antisense DNP-RNA inhibitor. The control compounds with sense or mismatched sequence were inactive. When c-myc transgenic mice were each treated with a single dose of the antisense RNA inhibitor, in vivo silencing of c-myc gene expression was observed for up to 72 hours by real-time RT-PCR. Similar treatment of c-myc transgenic mice with unmodified (native) homologous small interfering RNA (siRNA) had no effect on the mRNA concentration of the c-myc gene. Injection of this short antisense poly-DNP-RNA into mice did not induce the synthesis of DNP-binding immunoglobulins in the host. The observed in vivo gene silencing by this antisense RNA inhibitor suggests its possible use as a therapeutic agent for cancers involving the deregulation of c-myc gene expression. PMID- 15788899 TI - Targeting and regulation of the HER-2/neu oncogene promoter with bis-peptide nucleic acids. AB - Antigene oligonucleotides have the potential to regulate gene expression through site-specific DNA binding. However, in vivo applications have been hindered by inefficient cellular uptake, degradation, and strand displacement. Peptide nucleic acids (PNAs) address several of these problems, as they are resistant to degradation and bind DNA with high affinity. We designed two cationic pyrimidine bis-PNAs (cpy-PNAs) to target the polypurine tract of the HER-2/neu promoter and compared them to an unmodified phosphodiester triplex-forming oligonucleotide (TFO1) and a TFO-nitrogen mustard conjugate (TFO2). PNA1 contains a + 2 charge and bound two adjacent 9-bp target sequences with high affinity and specificity, but only at low pH. PNA2 contains a +5 charge and bound one 11-bp target with high affinity up to pH 7.4, but with lower specificity. The PNA:DNA:PNA triplex formed by these cpy-bis-PNAs presented a stable barrier to DNA polymerase extension. The cpy-bis-PNAs and the TFO-alkylator conjugate prevented HER-2/neu transcription in a reporter gene assay (TFO2 = PNA1 > PNA2 >> TFO1). Both PNAs and TFOs were effective at binding the target sequence in naked genomic DNA, but only the TFO-alkylator (TFO2) and the more cationic PNA (PNA2) were detected at the endogenous HER-2/neu promoter in permeabilized cells. This work demonstrates the potential for preventing HER-2/neu gene expression with cpy-bis-PNAs in tumor cells. PMID- 15788900 TI - Serum alters the uptake and biologic activity of CpG oligodeoxynucleotides in B cell chronic lymphocytic leukemia. AB - Immunostimulatory CpG-containing oligodeoxynucleotides (CpG ODN) have a number of effects on B cells, including upregulation of immunogenic molecules, and, therefore, appear attractive as potential components of immunotherapy for B cell chronic lymphocytic leukemia (B-CLL). Previous in vitro studies investigating the effect of CpG ODN on B-CLL cells used serum-low conditions and did not account for the longer-half life of CpG ODN in vitro. The present study was designed to explore how the presence of serum and exposure time affect CpG ODN-mediated changes on B-CLL cells. The optimal concentration for CpG ODN-mediated effects in the presence of 100% serum or plasma was higher (10-20 microg/ml) than for serum low conditions. Maximal CpG ODN-mediated effects required the presence of ODN for no longer than 3 hours. The inhibition of CpG ODN-mediated effects by serum correlated with lower uptake of ODN into B-CLL cells in the presence of serum. A threshold effect on biologic response was observed, with a given amount of ODN internalized, resulting in phenotypic changes. In conclusion, systemic short-term application of CpG ODN appears to be sufficient to induce phenotypic changes, but higher doses of CpG ODN than previously thought may be necessary because of inhibition of their uptake by serum. PMID- 15788902 TI - Do anti-inflammatories have a role in wound healing? PMID- 15788910 TI - Meeting the challenge of skin tears. PMID- 15788911 TI - 2005 HCPCS changes impact wound care. PMID- 15788912 TI - Management of a complex achilles wound. PMID- 15788913 TI - Acknowledged funding sources in pressure ulcer literature: a systematic review. AB - OBJECTIVE: To identify pressure ulcer research funding sources. DESIGN: A systematic review of the acknowledgment sections of pressure ulcer literature published between 2000 and 2002. RESULTS: Sixty-four papers were identified that acknowledged funding sources. Governmental funding sources were acknowledged in 53% (n = 34) of these papers, followed by manufacturers (30%, n = 19), consumer/professional organizations (17%, n = 11), provider organizations (11%, n = 7), philanthropic organizations (11%, n = 7), and academic institutions (9%, n = 6). CONCLUSION: Although various governmental organizations fund most pressure ulcer-related projects, this systematic review found that many other sources of funding are available. Researchers are encouraged to explore multiple sources for funding clinical research. PMID- 15788914 TI - Predicting pressure ulcer risk: comparing the predictive validity of 4 scales. AB - OBJECTIVE: To compare the predictive validity of 4 pressure ulcer risk assessment tools. METHODS: Prospective clinical design in which 230 subjects free of pressure ulceration on admission were assessed using the Braden, Gosnell, Norton, and Waterlow scales within 48 hours of admission. Subjects' skin condition was assessed once every 24 hours for a minimum of 14 days to identify any skin breakdown. RESULTS: Based on Youden's index, the Gosnell Scale had better predictive validity in identifying patients at risk for pressure ulcer development (J = 68%). The other scales did not predict individuals at risk with high accuracy, despite having high sensitivity and specificity. CONCLUSION: Numerous pressure ulcer risk assessment tools have been developed, but sufficient evidence for using one tool over another does not exist. In this study, the Gosnell Scale was found to be more appropriate for application in patients with neurologic and orthopedic conditions. CLINICAL RELEVANCE: Being able to predict which patients are at risk for pressure ulcers can assist practitioners in tailoring care to prevent unnecessary complications and suffering, as well as reduce costs. PMID- 15788915 TI - What does the wound say? Why determining etiology is essential for appropriate wound care. AB - PURPOSE: To provide physicians and nurses with an overview of factors to consider in determining wound etiology. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in understanding how determining wound etiology affects wound care. OBJECTIVES: After reading the article and taking the test, the participant should be able to: 1. Describe the etiology and characteristics of pressure ulcers, skin tears, and diabetic ulcers. 2. Describe the etiology, characteristics, and management of venous and arterial ulcers. 3. Identify factors needed to determine the etiology and management of specific atypical wounds. PMID- 15788917 TI - Perceptions of human immunodeficiency virus (HIV) testing services among HIV positive persons not in medical care. AB - BACKGROUND AND OBJECTIVES: Human immunodeficiency virus (HIV) counseling, testing, and referral (CTR) are provided in a wide variety of settings. GOAL: To compare, by test setting, the perceptions of the testing experience among HIV positive persons who were not receiving medical care. DESIGN: A baseline questionnaire was administered at enrollment into the Antiretroviral Treatment Access Study by the use of audio computer-assisted self-interview. RESULTS: Of 316 respondents, 27% reported that the counselor did not spend enough time with them and 22% that the counselor did not answer all questions. The odds were higher that persons in the following settings, compared with those at HIV test sites, would report that the counselor did not spend enough time with them: office of private physician or health maintenance organization (HMO) (adjusted odds ratio [AOR], 5.24; 95% confidence interval, 1.26-21.7), jail (AOR, 5.10; 95% CI, 1.06-24.6), and emergency room (ER) or hospital overnight visit (AOR, 2.93; 95% CI, 1.15-7.44). Similarly, the odds were higher that persons in the following settings compared with those at HIV test sites would report that the counselor did not answer all questions: office of private physician or HMO (AOR, 9.62; 95% CI, 2.22-41.7), jail (AOR, 7.87; 95% CI, 1.50-41.4), and ER or hospital overnight visit (AOR, 3.32; 95% CI, 1.11-9.90). CONCLUSION: Further training and quality assurance in HIV CTR may be needed in some test settings. PMID- 15788918 TI - Postexposure prophylaxis after sexual assaults: a prospective cohort study. AB - OBJECTIVE: The objective of this study was to evaluate HIV postexposure prophylaxis (PEP) protocol in rape victims. STUDY: The victims were assigned to 1 of 3 categories, according to the severity of exposure (I-low, II-moderate, III high). HIV PEP was provided to victims in groups II (Zdv + 3TC) and III (Zdv + 3TC + PI) until 72 hours after exposure. The follow-up was 6 months. RESULTS: From May 1997 to October 2001, 347 victims were attended. PEP was offered to 278 victims (141 in group II and 137 in group III). Side effects were more common in group III (P <0.01). No seroconversion was diagnosed in the 180 victims that completed the follow-up. Univariate analysis showed that the schooling level, knowledge of the aggressor's HIV status, and the use of PEP were associated with compliance. CONCLUSIONS: Triple therapy was associated with side effects, which suggested that drug regimes should be reviewed. The variables related to a high risk of HIV transmission were also significant for compliance. PMID- 15788919 TI - The re-emergence of syphilis in the United Kingdom: the new epidemic phases. AB - OBJECTIVE: The objective of this study was to characterize the resurgence of infectious syphilis in the United Kingdom between 1997 and 2003. STUDY: The authors conducted a retrospective analysis of routine surveillance data from genitourinary medicine clinics and data collected through enhanced surveillance. RESULTS: Between 1997 and 2002, diagnoses of primary, secondary, and early latent syphilis made at genitourinary medicine clinics increased by 213% in heterosexual males, 1412% in men who have sex with men (MSM), and 22% in females. These increases have been driven by a series of outbreaks, the largest of which were seen in Manchester (528) and London (1222) up to the end of October 2003. All the outbreaks have been geographically localized and the majority of cases occurred in MSM. A high percentage of concurrent HIV infection was reported, and oral sex was often reported as a route of transmission. CONCLUSIONS: Syphilis has re emerged in response to behavior change, probably driven by changes in the HIV epidemic. The future course of the epidemic is difficult to predict and control remains elusive. PMID- 15788920 TI - Vaginal swabs versus lavage for detection of Trichomonas vaginalis and bacterial vaginosis among HIV-positive women. AB - OBJECTIVES: Cervicovaginal lavage (CVL) is often used for research and may be easier and more accurate than vaginal swabs as a specimen collection method. GOAL: The goal of this study was to compare (CVL) with vaginal swabs for the detection of bacterial vaginosis (BV) and Trichomonas vaginalis (TV). STUDY: CVL and vaginal swabs were collected from 216 HIV-infected women. Clinical assessments were made using wet mount for TV and Amsel's criteria for BV through CVL and swab collection methods. Laboratory gold standards used were Nugent's criteria for BV and InPouch (Biomed Diagnostics, San Jose, CA) culture for TV collected by swab. RESULTS: The prevalence by gold standards for BV was 49.3% and for TV was 25.2%. Sensitivities for direct microscopy versus culture for TV were 72.2 for CVL and 52.8 for vaginal swab (P <0.05). Sensitivities for Amsel's versus Nugent's criteria for BV were 36.2 for CVL and 34.0 for vaginal swab (P <0.80). Kappa scores of agreement between CVL and vaginal swabs for BV and TV were excellent for both. CONCLUSION: CVL was comparable to vaginal swabs as a specimen collection method for these 2 lower genital tract infections and may be superior for the diagnosis of TV. PMID- 15788921 TI - Improving the accuracy of syndromic diagnosis of genital ulcer disease in Malawi. AB - OBJECTIVES/GOAL: Most resource-poor settings rely on syndromic criteria to diagnose genital ulcer disease (GUD). However, the etiologic pathogens of GUD vary temporally and geographically, and current criteria may not reflect changes in the prevalence of specific pathogens. STUDY: In 1999, we estimated the prevalence of Treponema pallidum (Tp), herpes simplex virus (HSV), and Haemophilus ducreyi (Hd) in Malawi. We then used regression coefficients of independent correlates of HSV and Hd to develop weighted diagnostic algorithms, in which weights were beta-coefficients corresponding to each factor. RESULTS: Overall, a decrease in the proportion of sexually transmitted disease attributable to GUD was noted in 7 years. Thirty-five percent were attributable to HSV, 30% to H. ducreyi, and 4% to T. pallidum. Areas under the receiver operating characteristic curves for weighted and unweighted HSV diagnostic algorithms were 67.6% and 66.5%, respectively. There was no significant difference in the explanatory performance of the weighted and unweighted algorithms. CONCLUSIONS: Unweighted algorithms can therefore be used to improve diagnostic accuracy of GUD. PMID- 15788922 TI - Acceptability of adult male circumcision for sexually transmitted disease and HIV prevention in Zimbabwe. PMID- 15788923 TI - Street youths are the only high-risk group for HIV in a low-prevalence South American country. AB - OBJECTIVES: To measure HIV prevalence in various subpopulations in Bolivia. DESIGN: In 2002 in Cochabamba, we offered voluntary counseling and testing to homeless street youths, registered and unregistered commercial sex workers, truck drivers, and prisoners. We examined surveillance data of pregnant women and blood donors. RESULTS: Among street youths over 15, overall HIV prevalence was 3.5% (11/313), higher among those recruited in the street, lower among those recruited in centers for homeless; prevalence was 0.6% (2/334) and 0.5% (1/189) in female registered and nonregistered sex workers, respectively, and below 0.3% in all other groups. All HIV cases were attributed to sexual transmission. CONCLUSION: In a low-prevalence setting where intravenous drug use is uncommon, street youths are a threat for the expansion of the HIV epidemic. We argue that HIV prevention in this population requires a comprehensive approach to their health and social problems. PMID- 15788924 TI - Trends in clinic visits and diagnosed Chlamydia trachomatis and Neisseria gonorrhoeae infections after the introduction of a copayment in a sexually transmitted infection clinic. AB - BACKGROUND: To meet their sexually transmitted infection (STI) control mission, STI clinics most often offer services at no or minimal cost to clients. However, there is little knowledge about the effects of charging service fees on clinic attendance and STI identification. As a result of budget shortfalls, a clinic fee of 15 US dollars for Denver residents (up to 65 US dollars for nonresidents) was introduced in the Denver Metro Health (STI) Clinic (DMHC) in December 2002. We evaluated the effects of the fee on clinic utilization and diagnosed Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) infections. METHODS: Using the DMHC computerized medical record system, we compared clinic visits and CT/GC diagnoses between the first 3 quarters of 2002 and 2003. We also compared CT/GC cases reported by DMHC with those reported by other Denver providers during 2002 and 2003. RESULTS: Compared with 2002, there were 3250 fewer visits (-28.5%) to DMHC in 2003 with no variance across quarters. CT diagnoses declined by 427 cases (-28.1%), disproportionately affecting women (-40%) and those under 20 (-42%). In addition, there were 332 fewer GC diagnoses (-38.1%) with no clear demographic preponderance. Although there were some decreases in non-DMHC CT/GC reports, the ratio of DMHC to non-DMHC reports declined from 0.42 to 0.33 (-21.4%) for CT and from 0.94 to 0.61 (-35.1%) for GC. CONCLUSIONS: Even a modest fee for service appears to have a major impact on the use of STI clinic services and may result in a significant effect on the ability to diagnose CT/GC infections, especially among those at highest risk. PMID- 15788925 TI - Sexually transmitted infections and other health conditions of women entering prison in Georgia, 1998-1999. AB - OBJECTIVES: To assess health needs of women entering the Georgia prison system, prevalence of pregnancy and sexually transmitted infections was estimated. STUDY: Results of admission screening tests of women entering the Georgia prison system in 1998 to 1999 were abstracted retrospectively from prison records. RESULTS: Of 3636 women whose data were abstracted from prison records, 4.3% were pregnant and 8.2%, 4.0%, 5.9%, and 0.7%, respectively, had positive screening tests for trichomoniasis, HIV, chlamydia, and gonorrhea; 19.5% had at least 1 of those conditions. HIV prevalence was higher among inmates who were black or had a rapid plasma reagin test for syphilis reactive at > or =1:8 dilutions (6.0%, 15.8%, respectively) than others (1.3%, 3.7%; P < 0.001). CONCLUSION: Inmates in this study had high rates of sexually transmitted infections and many were pregnant. Black inmates were at higher risk for HIV and high rapid plasma reagin titers than white inmates or other routinely tested Georgia female populations. PMID- 15788926 TI - The algebra of condoms and abstinence. PMID- 15788927 TI - Chlamydia trachomatis and Neisseria gonorrhoeae prevalence and coinfection in adolescents entering selected US juvenile detention centers, 1997-2002. AB - BACKGROUND: Juvenile detention centers offer public health practitioners an opportunity to gain access to large numbers of adolescents at risk for chlamydia and gonorrhea. GOAL: To describe the prevalence and coinfection of chlamydia and gonorrhea among adolescents in 14 US juvenile detention centers from 1997 to 2002. STUDY: We calculated the prevalence of chlamydia and gonorrhea in males and females, stratified by race/ethnicity, age group, and site. We also calculated the proportion of adolescents with chlamydia that were coinfected with gonorrhea and the proportion of those with gonorrhea that were coinfected with chlamydia. RESULTS: The prevalence of chlamydia was 15.6% in 33,619 females and 5.9% in 98,296 males; gonorrhea prevalence was 5.1% in females and 1.3% in males. Of females with gonorrhea, 54% were coinfected with chlamydia, and 51% of males with gonorrhea were coinfected with chlamydia. CONCLUSIONS: Chlamydia and gonorrhea prevalence was very high in females in all project sites. In males, chlamydia prevalence was high in some areas; however, gonorrhea prevalence was substantially lower. These prevalence data justify screening for chlamydia and gonorrhea among female adolescents in juvenile detention centers nationally. PMID- 15788928 TI - The prevalence of chlamydia, gonorrhea, and trichomonas in sexual partnerships: implications for partner notification and treatment. AB - BACKGROUND: Treatment of sex partners by patient-delivered partner therapy (PDPT) may prove to be an effective strategy in reducing reinfection and preventing the sequelae of sexually transmitted infections (STIs). However, limited data exists regarding STIs within sexual partnerships (dyads). OBJECTIVE: The objective of this study was to determine the prevalence of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (GC), and Trichomonas vaginalis (TV) in sexual dyads to estimate the potential yield and limitations of PDPT. METHODS: Male and female STI clinic attendees were invited to participate. Index subjects and partners were interviewed and tested for CT, GC, and TV. All partners were sought regardless of infection status of the index subject. RESULTS: Of 210 dyads, the prevalence in index subjects was CT, 46%; GC, 18%; and TV, 14%. Considering the partners of 72 CT-only-infected index subjects, 57% had CT, 6% had GC, and 11% had TV. Considering the partners of 35 index subjects with GC or GC-CT coinfection, 57% had GC and/or CT; however, in 20% of partners, unsuspected TV was present. Among 74 dyads with uninfected index subjects, 26% of partners had an STI. Among the partners of 19 index subjects with TV only, 11% had CT, 5% had GC, and 37% had TV. CONCLUSION: In our clinic population, a substantial number of partners had infections different from or in addition to those infections in the index. Many of these infected partners would not be diagnosed and treated using PDPT. Partners of index attendees without detected infection were at high risk (26%) for STI, mostly CT. PMID- 15788929 TI - Undercover cameras show nursing in a poor light. PMID- 15788930 TI - Supporting people with long-term conditions. PMID- 15788931 TI - Male obesity: a gender-specific approach to nurse management. AB - Android or male obesity is increasing. With greater understanding of the causes, development and outcomes of being obese and overweight, the nurse can begin to help men by using a gender-specific approach towards challenging this increasing global epidemic. This article provides a definition of overweight and obesity using the World Health Organization classification. The prevalence of obesity among men in the UK is outlined, and inequalities are highlighted and discussed. Some men from certain social, economic and ethnic groups are predisposed to developing obesity or becoming overweight. The human and financial implications of obesity and overweight are described and the healthy outcomes associated with the disease are discussed. Methods used to prevent obesity and overweight, such as information giving, pharmacological intervention and bariatric surgery, are outlined. Finally, a call is made for nurses to address male obesity from a gender perspective taking into account the attitudes, hopes and aspirations of men from various social, ethnic and economic backgrounds. Long-term local and national strategic policy is needed if this upward trend in obesity and overweight is to be addressed effectively. Joined-up intergovernmental policy making approaches are advocated. PMID- 15788932 TI - The management of obesity: the role of the specialist nurse. AB - Obesity is a global problem, independent of age. The numbers of obese individuals are now reaching epidemic proportions around the world. This is contributing to the risk of inherent comorbidity. The pathophysiology of obesity, although widely debated, is still unclear with suggestions that multiple genetic mutations may have a key role in the development, but as yet no one genetic mutation is felt to be entirely responsible. Biochemical manifestations such as diabetes may play a role. The first goal of management of the obese patient will involve dietary and behavioural modification and a programme of physical exercise. In primary care settings, nurses are suitably placed to assess and manage obese patients (National Institute for Clinical Excellence (NICE), 2001a). The nursing profession needs to rise to the challenge and prepare nurses for a specialist role in obesity management. PMID- 15788933 TI - Measuring blood pressure using the mercury sphygmomanometer. AB - Automated blood pressure devices have been embraced by clinicians for their convenience and ease of use. However, the pressure to remove mercury-containing medical devices is leading to the demise of the mercury sphygmomanometer and the loss of manual blood pressure measurement skills. The article argues that there is still a place for the mercury sphygmomanometer, particularly when a clinical decision is based on the blood pressure readings. It presents a review of the available literature to explain the rationale for the use of manual sphygmomanometry and gives a step-by-step guide to the procedure with current supporting evidence. There is a brief discussion of special considerations when dealing with certain client groups. Normal blood pressure ranges for adults and children have been included along with a definition of hypertension for both groups. PMID- 15788934 TI - Exploring the law regarding the making and execution of wills. PMID- 15788935 TI - Explanatory models of diabetes among Asian and Caucasian participants. PMID- 15788936 TI - Senior nurse who repeatedly slept while on night duty. PMID- 15788937 TI - Improving the continuing care for individuals with spinal cord injuries. AB - The author of this article responded to a need to address a service deficit for individuals with spinal cord injury, by setting up a nurse-led outpatient clinic to review, assess, advise and help patients develop problem-solving strategies to enable them to maintain their level of independence in the community, prevent unnecessary admissions and minimize the development of more serious complications that arise as a consequence of ageing with a spinal cord injury. The clinic sessions provided an effective service which demonstrated its value by achieving the following outcomes: provided a practical problem-solving approach to changes in physical function; prevented unnecessary hospitalization; identified issues that required referral for additional specialist/services. Unfortunately, despite its short-term success, there were a number of issues (staffing and funding) which prevented the continuation and further development of the service. However, although not yet currently published, the key themes of the National Service Framework (NSF) for individuals with neurological conditions advocated the type of approach to patient care that has been reflected within this clinic (lifelong support from specialist practitioners and person-centred care). Hopefully the publication of such an influential document will support the development of future initiatives within the specialist field. PMID- 15788939 TI - Flocath quick: a new hydrophilic-coated catheter for intermittent use. AB - The following article looks at the new Flocath quick intermittent catheter, which as been developed with the aim of improving the quality of lives of clients and carers who undertake intermittent catheterization. Teleflex Medical (formerly Rusch UK) provides an extensive range of intermittent catheters in both male and female lengths, as well as a range of indwelling catheters widely used throughout the community, hospitals and private nursing facilities. PMID- 15788938 TI - All preregistration students should develop skills in learning disabilities. AB - In the light of ongoing service developments and government legislation it is important that preregistration nurses from all branches have the opportunity to develop their confidence and skills in the care of people who have learning disabilities. This article outlines the development of strategies employed by one institute of higher education in order to reflect the principles of "Valuing People" from a health perspective in non-learning disability nursing programmes. The particular challenges that have been encountered in the implementation of these strategies over the last 3 years are discussed, as are issues faced in the evaluation of the effectiveness of such strategies. PMID- 15788942 TI - Identification of skin cancers 2: malignant lesions. AB - With the incidence of skin cancers doubling in the last 20 years, it is becoming more and more important for health professionals to be able to identify these dangerous malignancies. In this, the second of two articles, the author discusses the three main types of malignant cutaneous lesion, and provides information on prevention that can be passed on to patients of all ages. PMID- 15788940 TI - Nurses are overtired and not getting enough sleep. PMID- 15788943 TI - An overview of constipation management for community nurses. AB - Constipation is thought to affect nearly one-fifth of the population, and can arise for a multitude of reasons, some physical, some psychological. Nurses must therefore approach constipation in the awareness of the range of contributory factors and consequently the range of treatment options available. PMID- 15788944 TI - Getting the most out of your clinical placement in district nursing. AB - The community placement is a vitally important part of any nurse's training, and the student has a responsibility to be proactive in learning from it. To this end, Northumbria University has pioneered a portfolio of learning opportunities (POLO) to provide students with a structure overview of the opportunities available to them during a placement and how to make the most of them. The POLO also assists mentors in framing their support of the student. In this article, a practice placement facilitator outlines the structure of a POLO, and argues for a more coherent approach to student placements. PMID- 15788945 TI - Urinary catheter-related infection: an education programme for users. AB - While infection control has increased in profile both clinically and politically, it has also become embedded in clinical governance frameworks of NHS Trusts. Over the last few years the Department of Health has produced a number of documents in an attempt to highlight the increasing risk of healthcare associated infection. Today, the need for education as a key component in health care has been recognized as well as the need for patients and carers to receive information and be trained in, for example, technique of hand decontamination and catheter management following discharge from hospital. The objective of the education programme outlined in this article was to work alongside government guidelines in an attempt to improve education and understanding about infection control and management of infection in the catheterized patient. PMID- 15788946 TI - Learning disabilities health check project. PMID- 15788947 TI - Patient empowerment in primary care: evaluating the expert patient. PMID- 15788948 TI - Representing "grass roots" nurses at the top of the management ladder. PMID- 15788949 TI - Developing motivation and empowerment in diabetes care. AB - Management of incurable chronic diseases, such as diabetes, relies to a great extent on the active participation of the individual with the disease. Yet a diagnosis of diabetes can have a significant psychological impact on someone which affects his or her motivation to participate. Management of the disease therefore requires a different approach to care than the curative approach which nurses are taught. This article examines techniques that can be used to engage people in their own care, increasing motivation and empowerment and leading to greater self-management of their condition. PMID- 15788950 TI - Human rights and district nursing practice. AB - The main provisions of the European Convention on Human Rights were incorporated into UK law in the Human Rights Act 1998. Human Rights were described by Lord Hoffman in 'Matthews v Ministry of Defence' [2003] as the rights essential to the life and dignity of the individual in a democratic society. The fundamental nature of the rights demand that district nurses must inform their practice with a clear understanding of the main provisions of the Act and how they apply to health care. PMID- 15788951 TI - Leading an empowered organization (LEO): does it work? AB - Leading an empowered organization (LEO) is a three-day course that has been delivered to over 40,000 front-line managers in the NHS, among them many district nurses. The aim of this review was to identify and assess all published literature about LEO; to critically appraise such evidence relating to the impact of LEO on participants' leadership behaviour and to critically appraise any evidence of the need for follow-up activities to sustain participants learning. Because of the limited quantity and quality of research literature (n = 6) it was considered legitimate to also include discussion papers (n = 2) and descriptive papers (n = 4) in the review. All papers are reviewed using a framework devised by the author broadly based on the critical appraisal checklist for qualitative research (Greenhalgh and Donald, 2000). From the available evidence it can be concluded that the LEO programme does have a positive impact on the leadership behaviour of participants, and that follow-up activities are valuable to consolidate and sustain participants' learning. PMID- 15788952 TI - Making information technology work. PMID- 15788954 TI - Betting on dopamine. PMID- 15788955 TI - Neurocognitive function in schizophrenia at a 10-year follow-up: a preliminary investigation. AB - INTRODUCTION: A wealth of evidence indicates that neurocognitive deficits are evident in patients with schizophrenia at both illness onset and after many years of treatment. Little is known regarding if or how these deficits change during the lifespan. The goal of the study was to evaluate changes in full-scale intelligence quotient and neurocognitive test performance over a 10-year interval in patients with schizophrenia. METHODS: Twelve patients were administered the Wechsler Adult Intelligence Scale-Revised as a measure of intellectual function and a neuropsychological test battery including measures of attention, verbal and non-verbal memory, language, visuospatial function, problem-solving, and motor function at entry to the study and at a 10-year follow-up. RESULTS: With the exception of performance on a measure of speeded motor sequencing, there was no significant decline in any of the measures at 10-year follow-up. Results from a measure of sustained auditory attention showed improvement at follow-up. DISCUSSION: These data support a neurodevelopmental model of schizophrenia for young adult to middle-age patients by suggesting that neurocognitive deficits that emerge either before disease onset or early in the course of the illness remain stable as the patient ages. CONCLUSION: Overall, measures of intelligence quotient, as well as specific neurocognitive skills, do not decline over a 10 year period in at least a subgroup of patients with schizophrenia. PMID- 15788956 TI - Genetic factors in neuromuscular pain. AB - Recent evidence suggests that fibromyalgia, a chronic widespread pain condition and related syndromes (chronic fatigue syndrome, irritable bowel syndrome, etc.) may share heritable pathophysiologic features. We review the recent literature on genetic and familial factors found to participate in the pathogenesis of these syndromes, specifically fibromyalgia, including evidence suggesting that serotonin- and dopamine-related genes may play a role in the pathogenesis of these illnesses. The importance of environmental factors triggering these conditions in predisposed individuals is also discussed. PMID- 15788957 TI - Understanding neuropathic pain. AB - Neuropathic pain is defined as a chronic pain condition that occurs or persists after a primary lesion or dysfunction of the peripheral or central nervous system. Traumatic injury of peripheral nerves also increases the excitability of nociceptors in and around nerve trunks and involves components released from nerve terminals (neurogenic inflammation) and immunological and vascular components from cells resident within or recruited into the affected area. Action potentials generated in nociceptors and injured nerve fibers release excitatory neurotransmitters at their synaptic terminals such as L-glutamate and substance P and trigger cellular events in the central nervous system that extend over different time frames. Short-term alterations of neuronal excitability, reflected for example in rapid changes of neuronal discharge activity, are sensitive to conventional analgesics, and do not commonly involve alterations in activity dependent gene expression. Novel compounds and new regimens for drug treatment to influence activity-dependent long-term changes in pain transducing and suppressive systems (pain matrix) are emerging. PMID- 15788958 TI - The clenching-grinding spectrum and fear circuitry disorders: clinical insights from the neuroscience/paleoanthropology interface. AB - This review discusses the clenching-grinding spectrum from the neuropsychiatric/neuroevolutionary perspective. In neuropsychiatry, signs of jaw clenching may be a useful objective marker for detecting or substantiating a self report of current subjective emotional distress. Similarly, accelerated tooth wear may be an objective clinical sign for detecting, or substantiating, long lasting anxiety. Clenching-grinding behaviors affect at least 8 percent of the population. We argue that during the early paleolithic environment of evolutionary adaptedness, jaw clenching was an adaptive trait because it rapidly strengthened the masseter and temporalis muscles, enabling a stronger, deeper and therefore more lethal bite in expectation of conflict (warfare) with conspecifics. Similarly, sharper incisors produced by teeth grinding may have served as weaponry during early human combat. We posit that alleles predisposing to fear-induced clenching-grinding were evolutionarily conserved in the human clade (lineage) since they remained adaptive for anatomically and mitochondrially modern humans (Homo sapiens) well into the mid-paleolithic. Clenching-grinding, sleep bruxism, myofacial pain, craniomaxillofacial musculoskeletal pain, temporomandibular disorders, oro-facial pain, and the fibromyalgia/chronic fatigue spectrum disorders are linked. A 2003 Cochrane meta-analysis concluded that dental procedures for the above spectrum disorders are not evidence based. There is a need for early detection of clenching-grinding in anxiety disorder clinics and for research into science-based interventions. Finally, research needs to examine the possible utility of incorporating physical signs into Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition posttraumatic stress disorder diagnostic criteria. One of the diagnostic criterion that may need to undergo a revision in Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition is Criterion D (persistent fear circuitry activation not present before the trauma). Grinding-induced incisor wear, and clenching-induced palpable masseter tenderness may be examples of such objective physical signs of persistent fear-circuitry activation (posttraumatic stress disorder Criterion D). PMID- 15788959 TI - Antidepressant treatment of psychotic major depression: potential role of the sigma receptor. AB - Psychotic major depression is a severe condition that frequently proves difficult to-treat. The most effective traditional treatments (electroconvulsive therapy and combinations of antipsychotics with tricyclic antidepressants) are associated with significant side effects, and the use of tricyclic antidepressants alone is largely ineffective. Recent evidence has indicated that the selective serotonin reuptake inhibitors, either alone or in combination with antipsychotics, may provide a desirable alternative to traditional treatments. Among selective serotonin reuptake inhibitors, fluvoxamine has been the best studied and, somewhat surprisingly, has proven effective in several studies as a monotherapy without the need to combine with an antipsychotic. It is proposed that the apparent efficacy of fluvoxamine in psychotic major depression may be related to its unique property of high affinity for the sigma 1 receptor, which is thought to play a role in psychosis and in the action of some antipsychotic drugs. PMID- 15788960 TI - Sentinel node concept in esophageal surgery: an elegant strategy. PMID- 15788961 TI - Surgical oncotaxis--excessive surgical stress and postoperative complications contribute to enhancing tumor metastasis, resulting in a poor prognosis for cancer patients. AB - We investigated the relationship between surgical stress and tumor metastasis. The excessive surgical stress of a thoracolaparotomy enhanced tumor metastasis remarkably in an experimental model. We would like to propose that this phenomenon be termed "surgical oncotaxis". This effect has previously been attributed to some mechanisms of immunosuppression, excessive secretion of corticoids, and active oxygen production of granulocytes. An increase in lipid peroxide (LPO) in the liver was observed after a thoracolaparotomy, but a strong radical scavenger of a DL-alpha-tocopherol-L-ascorbic acid 2-0-phosphate diester (EPC-K1) restrained LPO levels in the liver and the effect of tumor metastasis in parallel. As clinical strategies for restraining the surgical oncotaxis, the control of any cytokine storm after surgery and/or the scavenging of active oxygen appears to be possible and hopeful, since it might be intermediated by cytokine. When pre-administration findings for EPC-K1 and methylpredonisolone were compared, EPC-K1 was found to be more suitable for restraining surgical oncotaxis, because serum LPO was only controlled with EPC-K1. The cytokine storm which occurs after surgery is augmented by a second stimulation, such as the administration of lipopolysaccharide, and no drug could control this well experimentally. Postoperative complications are a clinical model of a second stimulation (a so-called second attack). Our data showed the prognosis of a group with complications to be worse than that of a group without them even though no difference existed in the background of the esophageal cancer patients studied. Based on these results, safe surgery and the choice of minimally invasive surgery are the best ways to control surgical oncotaxis. Following a major surgical procedure, such as a thoracolaparotomy, the use of corticoids and/or radical scavengers can contribute to restraining surgical oncotaxis. PMID- 15788962 TI - Surgical technique of experimental lung transplantation in rabbits. AB - BACKGROUND: Although rabbits have often been used as an experimental model for the analysis of lung preservation, there are no reports of long-term survival after rabbit lung allotransplantation. The purpose of this study was to establish a lung allotransplantation model for the evaluation of acute lung rejection in the rabbit and to investigate the transplantation techniques in the rabbit. METHODS: Left unilateral lung allotransplantations were performed in 10 pairs of Japanese white male rabbits, weighing from 2.8 to 3.7 kg. Rabbits were divided into two groups. Group A rabbits (n=5) received Cyclosporine A (CsA) (20 mg/kg/day) orally for 5 days postoperatively, while Group B rabbits (n=5) received no CsA. All rabbits were sacrificed at the fifth postoperative day for histological examinations. RESULTS: Anastomoses of the pulmonary vein were achieved by using the atrial cuff technique. Satisfied blood flow was obtained in all arterial and venous anastomosis sites. Bronchial anastomosis was also well healed and all rabbits could maintain adequate spontaneous ventilation. In Group A, histopathology revealed that three cases were grade A0, one was grade A1 and the other was A2. In Group B, three cases lived for five days, and histopathology showed two cases were grade A2 and one case was grade A3. CONCLUSION: We established a left unilateral lung allotransplantation model in the rabbit and observed suppression of acute rejection of the transplanted lung by CsA. This study suggests that the rabbit is also an experimental model suited for the analysis of lung preservation as well as lung allotransplant rejection. PMID- 15788963 TI - Experimental study on myocardial protection by adjunct use of carperitide (hANP) in cardiac surgery. AB - BACKGROUND: In recent years, various beneficial roles of human atrial natriuretic peptide (hANP) have been demonstrated in the internal medicine and surgical fields. However, direct myocardial protection by hANP against myocardial ischemic reperfusion injury has been rarely investigated. Thus, we investigated it from aspects of cardiac surgery. METHODS: Twenty-four pigs underwent extracorporeal circulation and were divided into three groups: control group (treated with only cardioplegic solution after aorta clamping; cardioplegic arrest for 30 minutes followed by reperfusion for 60 minutes); low dose group (treated with cardioplegic solution and ANP (25 microg)); and high dose group (treated with cardioplegic solution and ANP (100 microg)). Blood and myocardial cGMP, myocardial Ca and ATP concentration were determined. Histological examinations were performed using an electron microscope. RESULTS: Blood and myocardial cGMP and myocardial ATP levels were significantly higher in the hANP treatment groups than the control group. Myocardial Ca concentrations were significantly lower in the hANP treatment groups than the control group. In electron microscopy, ischemic reperfusion injury was rarely observed in the hANP treatment groups. CONCLUSION: The study demonstrated that hANP improves ischemic reperfusion injury and suggested that hANP exerts direct myocardial protection against myocardial injury associated with cardiac surgery (cardioplegic arrest while cardiopulmonary bypass). PMID- 15788964 TI - Epicardial radiofrequency ablation on a beating heart: an experimental study. AB - PURPOSE: The effect of epicardial radiofrequency ablation (RFA) during normal heart beating was experimentally studied in order to establish safe and effective procedures for RFA. METHODS: Seven pigs weighing approximately 30 to 50 kg were used in this study. Fifty-one epicardial RFA lesions were created on both atria using a Cobra Cooled probe with continuous internal irrigation of a saline solution. The ablation temperature was fixed at 80 degrees C and the duration of the RFA in each case was 20, 30, 60 and 120 seconds. RESULTS: There was significant positive correlation between the right and left atria in wall thickness. Transmural coagulation was obtained in 69% of the total specimens, which decreased according to the increase of wall thickness especially over 3 mm. Transmural coagulation was seen in 64% of the specimens after RFA of less than 30 seconds, and 86% after ablation of >or=60 seconds. Occurence of 90% or deeper coagulation was higher in the right atrium than in the left one (97% vs. 78%). Right atrial rupture occurred in a region of 1 mm in thickness after ablation of 60 seconds. CONCLUSION: Further technical improvements associated with new instruments are indispensable to complete epicardial RFA procedures on a beating heart. PMID- 15788965 TI - Gastrointestinal complications in patients undergoing coronary artery bypass grafting. AB - PURPOSE: Gastrointestinal complications (GICs) such as gastroduodenal ulcer, enterocolitis, and ischemic colitis after coronary artery bypass grafting (CABG) are rare, but are associated with high mortality and morbidity. The present study was performed to detect risk factors and to investigate outcomes following GICs after CABG. METHODS: Between January 1992 and December 2001, 17 of 549 patients (3.1%) developed GICs after CABG with cardiopulmonary bypass, presenting with gastrointestinal bleeding due to gastroduodenal ulcer, enterocolitis, or ischemic colitis. We conducted a retrospective analysis of these patients. RESULTS: All patients required emergent treatment for hemorrhage by means of blood transfusion and endoscopic ablation and/or clipping. The following possible predictors of GICs were identified by logistic multivariate analysis: age over 70, diabetes mellitus (particularly insulin-dependent diabetes), history of cerebrovascular disease or history of renal failure and postoperative low output syndrome (LOS). CONCLUSION: Our results suggested that GICs after CABG with cardiopulmonary bypass are rare but can be lethal. Early diagnosis and prompt intervention can be difficult but are potentially life saving for patients in whom GICs develop. PMID- 15788966 TI - Surgical results in acute type A aortic dissection. AB - Currently international registry data present the patient mortality with acute type A aortic dissection managed non-surgically to be 58%, and managed surgically to be 26%. Many articles consistently report the hospital mortality exceeding 20% in western countries. Many factors, such as cardiac tamponade and dissection related organ malperfusion, contribute to hospital mortality and morbidity. In Japan, the number of patients enrolled in the annual reports has been increasing and the surgical results have been improving year by year. In-hospital mortality has decreased to less than 20% since 1999. Since the beginning of our aortic program, a total of 98 patients underwent emergency operations, and the operative mortality and in-hospital mortality were 5.1% and 6.1%, respectively. In a recent series since 2001, the operative and in-hospital mortalities were remarkably low; 2.8% and 3.2% respectively. We were able to benefit greatly by various innovative technologies which include open distal anastomosis using hypothermic circulatory arrest with antegrade cerebral perfusion, gelatin-resorcin-formaldehyde (GRF) glue, branched presealed Dacron graft, and antegrade arterial perfusion. Our tear oriented surgery could be justified in many patients in order to improve the surgical results. In patients with preoperative organ malperfusion, it is still challenging to improve the mortality and morbidity. PMID- 15788967 TI - Injury of an aberrant subclavian artery: a rare complication of video assisted thoracoscopic esophagectomy. AB - An aberrant right subclavian artery is an uncommon anomaly. When associated with esophageal cancer, it can cause diagnostic confusion as the symptoms are similar. If unrecognized and injured during esophageal surgery, it can lead to disastrous complications. We report a patient in whom this aberrant artery was injured during a thoracoscopic mobilization of the esophagus. The embryological and radiologic aspects of this anomaly and its clinical significance are discussed. Pre-operative diagnosis will require a high index of suspicion, as the clinical and radiological features are not specific. If injury occurs, an immediate vascular reconstruction will prevent limb ischemia and hence knowledge of this entity is of utmost importance. PMID- 15788968 TI - Node dissection for solitary interlobar node metastasis from renal cell carcinoma by VATS. AB - We present an unusual case of a patient with a right pulmonary interlobar node metastasis from renal cell carcinoma following nephrectomy. She underwent interlobar node dissection (ND) by video-assisted thoracoscopic surgery (VATS). Interlobar ND without lobectomy by VATS has not been reported until now in English literature. The retraction of the right intermediate bronchus is a useful technique during this procedure. PMID- 15788969 TI - Pulmonary sequestration associated with asymptomatic aspergillosis. AB - Intralobar pulmonary sequestration associated with asymptomatic aspergillosis is a rare case. We describe the case of a 65-year-old woman with intrapulmonary sequestration, anomalous systemic arterial supply to the left lower lobe and aspergillosis who underwent left lower lobectomy and ligation of an anomalous artery by Video-Assisted Thoracoscopic surgery (VATS). Pathological examination showed the parenchymal distortion and chronic inflammation. Aspergillus were found in the cyst. VATS lobectomy for intralobar pulmonary sequestration is a safe and valid procedure. PMID- 15788970 TI - Absent pulmonary valve associated with tetralogy of Fallot and complete atrioventricular septal defect: report of a case. AB - A male infant with an extremely rare combination of absent pulmonary valve, tetralogy of Fallot and atrioventricular septal defect presented without symptoms of respiratory distress or congestive heart failure. He underwent successful primary repair at the age of 5 months. The procedure consisted of double-patch repair of the atrioventricular septal defect and right ventricular outflow tract reconstruction with a monocusp transannular patch. Resection or plication of a dilated pulmonary artery was not required. The patient is doing well without any symptoms 5 years after repair. PMID- 15788971 TI - Mitral valve repair for mitral insufficiency due to infective endocarditis in a patient with idiopathic thrombocytopenic purpura. AB - A 63-year-old woman with an 18-year history of idiopathic thrombocytopenic purpura (ITP) was admitted with a persistent fever of unknown cause. Blood culture was positive for alpha-Streptococcus and echocardiography revealed severe mitral regurgitation and vegetation on the mitral valve. After antimicrobial therapy for six weeks, she underwent mitral valve repair using a Cosgrove ring. The platelet count increased and remained stable by perioperative treatment with intravenous high-dose gamma-globulin and platelet transfusion without steroids therapy or splenectomy. The hospital course was uneventful. Perioperative high dose gamma-globulin therapy and platelet transfusion for the cardiac operation were useful to increase and maintain the platelet count for an ITP patient complicated with infective endocarditis. PMID- 15788973 TI - Surgical treatment for innominate artery aneurysm with a coronary pulmonary artery fistula: a case report. AB - A 50-year-old woman complained of hoarseness and chest X-ray demonstrated a widening of the superior mediastinum. Computed tomography scanning and aortography demonstrated aneurysmal dilatation at the innominate artery. Coronary arteriography showed a coronary pulmonary artery fistula originating from the left anterior descending artery. The surgical procedure was prosthetic bifurcated bypass grafting from the ascending aorta to the right common carotid artery and right subclavian artery. During the period of innominate artery occlusion, the patient was cooled to 25 degrees C with selective cerebroperfusion and circulatory arrest. Pathologic diagnosis of the specimen was that of a true aneurysm with atherosclerotic plaque and calcification. The coronary pulmonary artery fistula was closed inside the pulmonary artery. There were no signs of neurologic deficit noted. Innominate artery aneurysm is uncommon and the surgical approach varies in each case. A coronary artery fistula is also an uncommon congenital malformation but has been recognized more often with the improvement in diagnostic techniques such as selective coronary arteriography. We performed successful surgical treatment for a rare case of innominate artery aneurysm with a coronary pulmonary artery fistula. PMID- 15788972 TI - Infective endocarditis during pregnancy: report of a case. AB - A 23-year-old woman, gravida 1, para, 0, was transferred at 29 weeks and 2 days gestation for management of infective endocarditis (IE). Based on vegetations attached to the mitral valve by echocardiography and positive blood cultures for Streptococcus mitis, the diagnosis of IE was made at the referring hospital. On admission to our hospital, echocardiography demonstrated a large vegetation and prolapse of the mitral valve with severe valve regurgitation. The fetal heart rate was 140/min. Ultrasound examination demonstrated an appropriate for gestational age 1,350 g fetus in vertex presentation. The day after admission, her membrane ruptured spontaneously, and she delivered a male infant by caesarean section at 29 weeks and 3 days of gestation. On the 42nd day of hospitalization, the mitral valve was reconstructed by quadrangular resection of the posterior commissure and annuloplasy with a prosthetic ring. A histological examination of specimens of the resected leaflets indicated that IE was active. One year later, the patient delivered a healthy infant without any complications by a caesarean section. The latest Doppler study demonstrated trivial mitral regurgitation and a mitral valve area of 2.5 cm2. PMID- 15788974 TI - Acquired arteriovenous fistula of the right forearm caused by repeated blunt trauma: a report of a rare case. AB - In acquired arteriovenous fistula (AVF), there is usually a history of penetrating injury and hemorrhage. We report a very rare case of an elderly man with acquired AVF of the right upper extremity without any history of penetrating trauma and hemorrhage, but with a history of repeated blunt trauma on his right forearm. Although no surgery was performed for the AVF, it was concluded close follow up would be prudent. PMID- 15788975 TI - A randomized comparison between rotational and standard coronary angiography. AB - AIM: Selective coronary angiography is nowadays the gold standard in the definition of coronary anatomy as well as the basis for percutaneous coronary interventions. However, the diagnostic accuracy of coronary angiography can be reduced if the number of angiographic views is inadequate or if the operator does not select appropriate projections. Rotational angiography (RA) has been proposed as an alternative technique in order to provide a more complete definition of coronary anatomy reducing, at the same time, radiation exposure and contrast medium dose. METHODS: We randomly assigned 31 eligible patients, undergoing diagnostic cardiac catheterization, to RA (n=16) and traditional angiography (TA, n=15). Total procedural time, fluoroscopy time, number of cine-runs, X-ray dose and contrast medium volume were recorded in both groups. RESULTS: There were no statistically significant differences between groups in age (59+/-5.8 vs 62.8+/ 9.6 years, P=ns), body mass index (26.7+/-3.5 vs 27.1+/-3.4 kg/m2, P=ns), total procedural time (20.6+/-6.6 vs 22.2+/-11.3 min, P=ns) and fluoroscopy time (3.9+/ 1.5 vs 4.9+/-1.8 min, P=ns). On the contrary, number of cine-runs, X-ray dose and contrast medium volume were significantly lower in RA patients as compared with TA patients (6.2+/-1.2 vs 9.7+/-2.1, P<0.01; 530.6+/-271.6 vs 831.2+/-343.9 mGy, P<0.05; 76.9+/-22.4 vs 102.9+/-26.4 ml, P<0.01, respectively). CONCLUSIONS: RA is safe and effective in defining coronary anatomy, leading to a significant reduction in radiation exposure and contrast medium volume. PMID- 15788976 TI - No reflow in patients undergoing primary angioplasty for acute myocardial infarction at high risk: incidence and predictive factors. AB - AIM: Percutaneous coronary intervention (PCI) is a consolidated therapeutic strategy for the treatment of acute myocardial infarction (AMI), but achieving a TIMI 3 flow does not always correspond to true tissue reperfusion. The aim of the study was to evaluate the incidence and predictive factors of no reflow in patients undergoing primary angioplasty (PCI) for AMI at high risk, in the setting of a provincial cardiological emergency network. METHODS: We retrospectively analyzed the ECGs of 360 consecutive patients undergoing primary PCI, between 2001-2004, recorded before and 90 min after the procedure, and compared them with the angiographic data. RESULTS: The patients were divided into 2 groups: group A (reperfused) with a >50% reduction in ST and group B (no reflow) with a <50% reduction in ST but a TIMI 3 flow in the epicardial vessel. The 2 groups were comparable in terms of mean age, sex, diabetes and AMI site. However, there were statistically significant differences between the groups in terms of precoronary time, Killip class IV, ejection fraction, mean leukocyte count, C-reactive protein, and the periprocedural administration of abciximab. Total mortality was 6%: 14% in group B vs 3% in group A. CONCLUSIONS: Our data show that a fair percentage of patients (24%) with a TIMI 3 flow after PCI during ST-elevation MI do not show ECG signs of effective reperfusion and have a higher in-hospital mortality rate. Precoronary time and the inflammatory phlogistic substrate are important independent predictors of no reflow. Pretreatment with abciximab, particularly if it is not periprocedural, can prevent the occurrence of no reflow. PMID- 15788978 TI - Coronary restenosis after implantation of drug-eluting stents. AB - Randomized trials comparing drug-eluting stents (DES) with bare-metal stents have shown that the former significantly reduce the incidence of angiographic and clinical restenosis into an unprecedented low, one-digit, range. However, post DES restenosis is not zero. Next to incomplete coverage with DES of the vessel segment injured by balloon angioplasty, factors such as stent underexpansion, stent overexpansion, and nonuniform distribution of stent struts have been associated with post-DES restenosis. Current evidence suggests that inadequate, though predominantly focal, delivery of the antiproliferative agent (sirolimus or paclitaxel) into the vessel wall is likely the common cause of post-DES restenosis. There is no consensus at present on how to treat post-DES restenosis. Long-term results reported to date on small numbers of patients undergoing interventional treatment for post-DES restenosis appear to be worse than outcomes observed after the index intervention, regardless of whether another DES was implanted or not, and warrant further study. PMID- 15788977 TI - Optimal antithrombotic treatment for percutaneous coronary intervention. AB - Recent years have witnessed significant advances in the percutaneous treatment of patients with atherosclerotic vascular disease. Anti-platelet and anti-thrombotic agents are routinely administered to minimize the risk of peri-procedural myonecrosis, stent thrombosis and other procedural complications. This article presents a current view of optimal adjunctive antithrombotic therapy for percutaneous coronary interventions (PCI), recognizing that optimal is a necessarily subjective label. This article focuses specifically on anticoagulant agents such as unfractionated heparin (UFH), the low-molecular weight heparins (LMWH), and direct thrombin inhibitors, and antiplatelet agents, such as aspirin, thienopyridines, and glycoprotein IIb/IIIa antagonists. It starts with a general discussion of anticoagulation and percutaneous intervention, followed by a summary of the modern-day view of the coagulation process. The mechanism of action of the individual agents is then presented, followed by some of the evidence base of recent clinical trials of anticoagulant and antiplatelet agents in PCI. Finally, we present summary recommendations for procedural anticoagulation in low risk, not-low risk, and high risk PCI, and list what we feel are appropriate doses for the agents employed. Ultimately, though, it is the individual interventional cardiologists who must decide for themselves exactly what constitutes optimal antithrombotic therapy for PCI. PMID- 15788979 TI - New pharmacological protocols to prevent or reduce contrast media nephropathy. AB - Contrast media associated acute renal failure represents the third cause of in hospital renal function deterioration after decreased renal perfusion and post operative renal insufficiency. Although generally benign, this complication shows a mortality rate ranging from 3.8% to 64%, depending on the increase of creatinine concentration. The mechanism by which contrast-induced renal failure occurs is not well understood. Contrast agent-associated nephrotoxicity appears to be a result of direct contrast induced renal tubular epithelial cell toxicity and renal medullary ischemia. Furthermore, a key mechanism seems to be alteration in renal dynamics, probably caused by imbalances between vasodilator and vasoconstrictor factors, including the activities of nitric oxide, prostaglandins, endothelin and reactive oxygen species. Recommendations to prevent contrast-associated nephrotoxicity are: 1) periprocedural hydration, 2) use of a low osmolality contrast, and 3) limiting the amount of contrast agent. Recently, considerable interest has resulted from the preliminary positive data on the effectiveness of prophylactic administration of antioxidant compounds (such as acetylcysteine and ascorbic acid) and fenoldopam. PMID- 15788980 TI - How to use the C-reactive protein in cardiac disease? AB - Inflammation is an important contributor to atherothrombosis. The C-reactive protein (CRP) is not only an excellent biomarker of inflammation, but it is also a direct participant in atherogenesis. CRP consistently predicts new coronary events, including myocardial infarction and death, in patients with ischemic heart disease. The predictive value of CRP is, in the majority of the studies, independent of and additive to that of the troponins and its levels can be modulated by statins. Prospective observational studies show that moderately elevated levels of CRP are associated with an adverse cardiovascular prognosis among healthy individuals. The availability of high sensibility assays for CRP should provide a valuable tool for identifying patients at risk of cardiovascular events in primary prevention in conjunction with lowering LDL cholesterol and may also have utility in the treatment of acute coronary syndromes with percutaneous coronary intervention (PCI) therapy. High CRP levels, associated with a higher risk, should suggest a more aggressive medical therapy in the long term and also an aggressive and invasive therapy in the short term, including the use of GP IIb/IIIa inhibitors, high doses of statins, and when a PCI is necessary, provisional stenting. Finally, CRP will provide a readily accessible marker for further testing of the inflammatory hypothesis in atherosclerosis. PMID- 15788981 TI - Endovascular treatment of extra-cranial carotid artery bifurcation disease. AB - Over the past decade, carotid stenting with embolic protection has emerged as a viable alternative to carotid endarterectomy in patients with severe extra cranial bifurcation atherosclerotic disease. Several patient subsets -- high surgical risk, symptomatic, asymptomatic, etc.,-- have and are being studied in multi-center trials throughout Europe and the United States. Results of trials to date suggest that, in the high surgical risk cohorts, carotid stenting is statistically on par with endarterectomy with strong trends toward superiority in at least 2 trials SAPPHIRE and ARCHeR. In addition to these important ongoing clinical studies, physician training, reimbursement, and registry development still are in the early stages of evolution in the US. PMID- 15788982 TI - Renal artery stenosis: a review of therapeutic options. AB - Renal artery stenosis (RAS) has traditionally been under recognized in clinical medicine as a cause of secondary hypertension and as a culprit for progressive ischemic nephropathy. While it is well recognized that atherosclerotic RAS is a progressive disease, and that surgical revascularization may result in lowering of blood pressure and prevention of progression of nephropathy, the high morbidity and mortality associated with surgical revascularization has kept the enthusiasm for revascularization low. With the recent advances in renal artery stent revascularization, a procedure that can be accomplished with <1% major complication rate, 90-95% success rate and 10-15% restenosis rate, multiple studies have reported the salutary hemodynamic benefits and increased awareness of prevalence of RAS in patients with vascular disease. Multiple studies have reported sustained blood pressure control in 70-80% of patients, stabilization of renal function in a similar percentage of patients and beneficial effect of renal artery stenting in patients with angina or heart failure. Further advances in therapy consisting of distal protection to diminish procedural atheroembolism and aggressive adjunctive medical therapy may allow clearly demonstrable benefits of renal artery stenting in prevention of ischemic nephropathy and reduction of cardiovascular events. PMID- 15788983 TI - Decrease in extra-cellular water compartment in overweight women after administration of oligomineral water associated to a hypocaloric diet regimen. AB - AIM: Alteration in body fluid distribution may have relevant implications in several human diseases. The use of oligomineral water is associated with increased diuresis and this could be extremely useful in subjects with water retention. The aim of the present study was to evaluate the effect of daily administration of 1.5 liters of oligomineral water over 4 weeks on body water compartment (extracellular and intracellular water [ECW and ICW]) in healthy overweight women who underwent a hypocaloric dietary regimen. METHODS: Twenty four healthy premenopausal women (aged 32+/-6 years; body mass index [BMI]: 27.4+/-2 kg/m(2)) were randomly divided into 2 groups and assigned to consume 1.5 liters daily of either tap (Group A) or oligomineral water over a 4-week period. Body composition was assessed by anthropometry and multifrequence bio-impedance analysis. RESULTS: Body weight and fat mass significantly decreased after dietary treatment in the 2 groups examined; although not statistically significant, a higher difference in body weight before and after treatment was found in Group B when compared to Group A (2.2+/-1 kg vs 1.7+/-0.8 kg; p=0.057). Body weight reduction was related to fat mass decrease, while no significant difference was found in fat-free mass among groups. TBW and ECW were reduced in both groups after treatment, while a greater reduction in both variables was found in Group B (p<0.01). The post-treatment ECW/TBW ratio was lower in Group B with respect to Group A (p<0.01). CONCLUSIONS: A supplementation with oligomineral water in overweight women, when associated to a hypocaloric dietary treatment, is able to reduce total and extracellular water content; this findings could have relevant implication in planning a successful dietary regimen for achieving and maintaining a normal body weight. PMID- 15788984 TI - Is under diagnosis of celiac disease compounded by mismanagement in the primary care setting?. A survey in the Italian Province of Brescia. AB - AIM: Celiac disease is under diagnosed in the primary care setting, mainly because of lack of awareness on the heterogenic manifestation of the disease. Furthermore, patients diagnosed at open access endoscopy may be mismanaged with incomplete dietary information. The aim of this paper was to evaluate prevalence and incidence in 1996 and 1997 for celiac disease in the Italian Province of Brescia and to obtain information on the extent of underdiagnosis and mismanagement. METHODS: The authors assessed the under diagnosis of celiac disease by relating the number of patients on gluten-free diet at 31 December 1997 (prevalent cases) to the expected number of patients in a population of 1,055,499 assuming 1/200 disease prevalence. Post-diagnosis management was assessed by questionnaire for all incident cases in the hospital practice and in the primary care setting. RESULTS: Five hundred and ninety-four prevalent cases were identified compared with an estimated disease prevalence of 5,000 cases, a figure corresponding to 8/9 under diagnosis. One hundred and thirty-five incident cases during 1996-1997 have been identified. Overall 80% of incident cases were symptomatic, but 40% only with the classical symptoms of malabsorption. Forty three of the 135 incident cases did not receive appropriate dietary education following diagnosis, a figure corresponding to 1:3 mismanagement, and all of them were diagnosed in the primary care setting at open access endoscopy. CONCLUSIONS: Our study indicates that under diagnosis of celiac disease in the primary care setting is compounded by disease mismanagement, a finding suggesting the need for increasing awareness not only on the heterogenicity of clinical manifestation but also on the appropriate dietary management of celiac disease. PMID- 15788985 TI - Diagnosis and treatment of chronic hepatitis B. AB - The diagnosis of chronic hepatitis B virus (HBV) infection is made using a combination of serological, virological, biochemical, and histological markers. The natural history of HBV infection can be divided into 3 phases: immune tolerant, immune active with chronic hepatitis B, and inactive carrier; patients in the immune active phase are candidates for antiviral therapy. The primary goal of therapy for chronic hepatitis B is suppression of viral replication, which has been shown to reduce hepatic necroinflammation and retard progression of hepatic fibrosis. Long-term suppression of serum HBV DNA is likely to reduce progression to cirrhosis and hepatic decompensation and may also decrease the risk of hepatocellular carcinoma. Current antiviral therapy for chronic hepatitis B includes interferon alpha, lamivudine and adefovir, with recent studies demonstrating good safety and efficacy of peginterferon and other nucleoside analogues that will soon become additional treatment options. In patients with HBeAg-positive chronic hepatitis B, antiviral treatment is indicated when the serum HBV DNA level is = or >10(5) copies/mL and the alanine aminotransferase (ALT) level is elevated, particularly greater than 2 times the upper limits of normal. For HBeAg-negative patients, the threshold for initiation of therapy is a HBV DNA level = or >10(4) in association with an elevated ALT level. The presence of at least moderate necroinflammation and the presence of fibrosis on liver biopsy, which is optional and not mandatory before therapy, may be useful in supporting the decision to initiate therapy. While undergoing therapy, patients require monitoring every 3 to 6 months to ensure compliance and to test for the development of resistance if an oral agent is used. Issues that remain controversial or need to be studied further are the necessity of a baseline liver biopsy, the HBV DNA and ALT thresholds for initiation of therapy, the optimal duration of antiviral therapy, selection of one agent over another, and the role of combination therapy. PMID- 15788986 TI - [Thyroid-intestinal motility interactions summary]. AB - Thyroid diseases may be related to gastrointestinal motility symptoms. Such symptoms can vary in degree and, sometimes, are the only clue of a thyroid disease or, at least, the first. The mechanism by which the thyroid hormones can influence gastrointestinal motility, even if not still completely elucidated, can be found in a synergism between a direct effect of the thyronins and an indirect effect mediated by cathecolamines on the muscle cell receptors. Neck discomfort and dysphagia are common findings in patients with thyroid diseases. Hyper- and hypothyroidism can impair esophageal motility, modifying pharyngo-esophageal structure and/or muscular function and interacting with the neuro-humoral regulation of the esophageal peristalsis. Oesophageal motility alterations, observed in patients affected by small non-toxic goiter, are less understandable. At the gastro-duodenal level, basic and postprandial electric rhythm alterations have been observed in hyperthyroid patients, often associated with delayed gastric emptying, too. In such patients, the autonomous nervous system dysfunction may even modify the neuro-hormonal mutual regulation (vagal influence decrease) of the gastro-duodenal myoelectric activity. Hypothyroidism may cause a delay of the gastric emptying too, but such pattern may also be related to an associated autoimmune disease or to an independent chronic modification of the gastric mucosa. Diarrhoea and malabsorption are common findings together with hyperthyroidism, whereas constipation is frequently observed in hypothyroidism. The clinically most demanding situation is certainly the secondary chronic intestinal pseudo-obstruction syndrome, which involves the bowel in most cases, but may also show up by means of a mega-small bowel or a mega-duodenum, or even all of the above. In conclusion it may be stated that: 1) thyroid diseases may be related to symptoms due to digestive motility dysfunction. 2) Any segment of the gastrointestinal trait may be involved. 3) The typical clinical manifestations of the thyroid illnesses may be borderline, missing or concealed by other intercurrent illnesses, especially in the elderly patients. 4) Motility-related digestive symptoms may conceal an underlying, easily misdetected, thyroid disease and must be therefore carefully analyzed. PMID- 15788987 TI - [The efficacy of dietetic intervention in multiple sclerosis]. AB - The involvement of nutritional factors in the etiopathogenesis of multiple sclerosis is currently being investigated. Notwithstanding the huge amount of data present in the literature, the possible etiological or protective role of nutrients with regard to the disease remain debatable. The epidemiological data suggest an association between multiple sclerosis and nutrition; the populations that take in a higher quantity of foods of animal origin (meat p<0.0001 and dairy products p<0.01) seem to be the most affected. A role of saturated fatty acids in the etiopathogenesis of myelinic damage has been hypothesised. Case control studies have identified certain foods that act as risk factors and others as protection in the onset of the disease. Some case control studies point to a time cause relationship between the intake of total calories (O.R. 2.03) and saturated fats (O.R. 1.88) and the incidence of multiple sclerosis; other prospective studies failed to confirm this hypothesis, negating the protective effect of a diet rich in anti-oxidant vitamins and polyunsaturated fatty acids. Intervention studies are discordant with respect to the effects of polyunsaturated fatty acid supplements on the course of the disease. In patients with a progressive chronic form of the disease, polyunsaturated fatty acids did not demonstrate any effect on the progression of the invalidating lesions. Interventions on patients suffering from an acute and remittent form have pointed to the significant effect of treatment with polyunsaturated fatty acids in slowing down the progression of lesions only in cases with a slight initial degree of disability or no disability (p=0.001) at all. They do, however, seem to confirm the hypothesis of an association between the gravity of the disease and consumption of saturated fats (p<0.05) and show an improvement trend in patients treated with polyunsaturated fatty acids, although the data are not statistically significant. PMID- 15788988 TI - [Nutrition in acute pancreatitis]. AB - Nutritional management during acute pancreatitis has the purpose to avoid a negative influence on the outcome and to preserve the morphofunctional integrity of the gut, preventing bacterial translocation. When the patient would start again normal nutrition after a period shorter than a week, thanks to the resolution of the clinical picture, and when the initial nutritional state of the patient is satisfactory, a particular nutritional support is not necessary. When the course of the disease is longer and the severity is higher, an early artificial nutritional support is advisable. Caloric needs thought to be useful are 25-30 kcal/kg/die; 40-60% of nutrient mixture should consist of carbohydrates and 20-30% of lipids. Proteins should be approximately 1.0-1.5 g/kg/die. On the basis of recent randomised, prospective clinical trials, enteral jejunal feeding is indicated as a first choice nutritional way, because of its ability to maintain the integrity of the intestinal barrier and its minimal effect on pancreatic secretion, acting significantly on inflammatory parameters and on prognostic markers. This procedure is not indicated when ileum is present and when it causes nausea, vomiting, abdominal pain and an increase of hepatic enzymes. In this case, parenteral feeding is an alternative. Hydroly-sated formulas, containing short peptides and a low percentage of long chain fat acids, are recommended. PMID- 15788989 TI - [Gastric dysplasia: classification and clinical characteristics]. AB - Gastric dysplasia is considered to be the morphologic expression of the penultimate stage in gastric carcinogenesis. The clinical importance has been underscored until the close association with gastric cancer was established. Nevertheless, similar lesions are considered as high grade dysplasia by Western pathologists and as carcinoma by Japanese pathologists, using the Japanese group classification. Padova classification, Vienna classification and Vienna revisited, try to overcome these differences; because of this disagreement therapeutic guidelines are also controversial and differences in the incidence of early stages of gastric cancer and its outcome were reported. PMID- 15788990 TI - Intrahepatic cholestatic jaundice related to administration of ranitidine. A case report with histologic and ultramicroscopic study. AB - Ranitidine may cause liver injuries ranging from transient, subclinical serum transaminases increase every 100-1,000 treated patients to cholestatic hepatitis in less than 1/100,000. Other H2-receptor antagonists are more dangerous: 11 toxic hepatitis cases have been reported as adverse effect after 1 year of marketed ebrotidine. A 75-year-old male with ischemic cardiopathy history was started on an 8 days treatment of oral ranitidine due to pirosis, without any other changes of therapy; 48 h after drug withdrawal, light-coloured stools, dark urine and icteric scleras developed. On hospital admission, 10 days later, physical examination showed slight hepatomegaly and severe jaundice with skin excoriations followed by serum mixed bilirubin further increase and aminotransferases activities mild rise. Total bilirubin peaked at 381.33 mmol/l (5.1-17.1) and progressively returned to normal, after discharge home, in 3 months and now, 1 year later, there is no sign of liver disease. Ultrasonographic biliary anomalies and the most frequent causes of liver damage were excluded. Liver biopsy confirmed ranitidine as the most likely cause of liver toxicity since histological and ultramicroscopical study revealed a drug-induced picture. We report a rare case of intrahepatic cholestasis jaundice related to ranitidine, a widely used drug. Diagnosis would need an ethically unacceptable rechallange test. PMID- 15788992 TI - Varicella-related deaths--United States, January 2003-June 2004. AB - During 2003 and the first half of 2004, CDC received reports of eight varicella related deaths. The age of the decedents ranged from 1 to 40 years. Six of the eight deaths occurred among children and adolescents aged <20 years. The cases were reported from Arizona (two), Maryland (two), Arkansas (one), New Hampshire (one), Ohio (one), and New York City (one). Six deaths occurred in unvaccinated persons. Vaccination status of the remaining two persons could not be determined. This report describes clinical data for three of the fatal varicella cases in children, reported from Arizona, Arkansas, and New York City; all three patients were susceptible and unvaccinated, but otherwise healthy. The three other children and adolescents, not described in detail in this report, were immunocompromised as a result of at least one preexisting condition. The findings in this report underscore 1) the importance of timely routine vaccination of children aged 12--18 months and catch-up vaccination of older susceptible children and adolescents according to current recommendations and 2) the need for timely and complete national varicella death surveillance. PMID- 15788991 TI - Pseudomonas bloodstream infections associated with a heparin/saline flush- Missouri, New York, Texas, and Michigan, 2004-2005. AB - On January 26, 2005, CDC was notified of four cases of Pseudomonas fluorescens bloodstream infection among patients at an oncology clinic in Missouri. All patients had received a heparin/saline flush to prevent clotting of indwelling, central venous catheters. The flushes were preloaded in syringes by IV Flush and distributed by Pinnacle Medical Supply (Rowlett, Texas). On January 31, a nationwide alert against use of all heparin or saline flushes preloaded in syringes by IV Flush was issued by the Food and Drug Administration; the company recalled these products. As of February 15, state and local health departments and CDC had identified a total of 36 Pseudomonas species infections in patients in four states who were administered the heparin/saline flushes from multiple lots. This report describes the ongoing investigation and provides recommendations for investigation and management of potential cases. PMID- 15788993 TI - Varicella surveillance in public elementary schools--Multnomah County, Oregon, 2002-2004. AB - Varicella vaccination of school-aged children reduces the number of varicella cases and lost days of school. In 1996, the Advisory Committee on Immunization Practices (ACIP) recommended routine vaccination of all children aged 12-18 months, catch-up vaccination of all susceptible children before age 13 years, and vaccination of susceptible persons who have close contact with persons at high risk for serious complications and susceptible persons at high risk for exposure. In 1999, ACIP updated these recommendations to include vaccination requirements for child care and school entry. Since 2000, in accordance with ACIP recommendations, varicella vaccination requirements have been phased in for Oregon children who have not had varicella before starting out-of-home child care, kindergarten, or seventh grade; elementary school children will be fully covered by school year (SY) 2006-07. To monitor changes in varicella incidence, Oregon Health Services (OHS) and Multnomah Education Service District (MESD) started routine, individual, case-based varicella surveillance in Multnomah County public elementary schools (kindergarten through 5th grade) beginning SY 2002-03. This report describes the surveillance system, the incidence of varicella during SY 2002-03 and SY 2003-04, and the results of active surveillance for unidentified cases during SY 2002-03. The findings indicate that the number of varicella cases has decreased in Oregon and that establishing public elementary school-based varicella surveillance is feasible and useful. PMID- 15788994 TI - Progress toward poliomyelitis eradication--Afghanistan and Pakistan, January 2004 February 2005. AB - Although poliomyelitis remained endemic in only six countries at the end of 2003, a resurgence of polio occurred in 2004, originating in Nigeria and resulting in the export of wild poliovirus (WPV) into the polio-free countries of western and central Africa. However, progress toward interrupting WPV transmission continued during 2004 in Afghanistan, India, and Pakistan, the only remaining countries in Asia where polio is endemic. This report summarizes progress toward polio eradication in Afghanistan and Pakistan during January 2004-February 2005 and indicates that, with continued support from national and local leaders, interruption of poliovirus transmission in both countries is feasible by the end of 2005. PMID- 15788995 TI - Elimination of rubella and congenital rubella syndrome--United States, 1969-2004. AB - In October 2004, CDC convened an independent panel of internationally recognized authorities on public health, infectious disease, and immunization to assess progress toward elimination of rubella and congenital rubella syndrome (CRS) in the United States, a national health objective for 2010. Since rubella vaccine licensure in 1969, substantial declines in rubella and CRS have occurred, and the absence of endemic transmission in the United States is supported by recent data: 1) fewer than 25 reported rubella cases each year since 2001, 2) at least 95% vaccination coverage among school-aged children, 3) estimated 91% population immunity, 4) adequate surveillance to detect rubella outbreaks, and 5) a pattern of virus genotypes consistent with virus originating in other parts of the world. Given the available data, panel members concluded unanimously that rubella is no longer endemic in the United States. This report summarizes the history and accomplishments of the rubella vaccination program in the United States and the Western Hemisphere and the challenges posed by rubella for the future. PMID- 15788996 TI - Compendium of measures to prevent disease associated with animals in public settings, 2005. AB - Multiple venues encourage or permit the public to come in contact with animals, resulting in millions of human-animal contacts each year. These settings include county or state fairs, petting zoos, animal swap meets, pet stores, zoologic institutions, circuses, carnivals, farm tours, livestock-birthing exhibits, educational exhibits at schools, and wildlife photo opportunities. Although multiple benefits of human-animal contact exist, infectious diseases, rabies exposures, injuries, and other human health problems associated with these settings are of concern. Rabid or potentially rabid animals in public settings can result in extensive public health investigation and action. Infectious disease outbreaks reported during the previous decade have been attributed to multiple organisms, including Escherichia coli O157:H7, Salmonella, Coxiella burnetti, Mycobacterium tuberculosis, and ringworm. Such incidents have substantial medical, public health, legal, and economic effects. This report provides standardized recommendations for public health officials, veterinarians, animal venue operators, animal exhibitors, visitors to animal venues and exhibits, and others concerned with disease-control and with minimizing risks associated with animals in public settings. The recommendation to wash hands is the single most important prevention step for reducing the risk for disease transmission. Other critical recommendations are that venues include transition areas between animal areas and nonanimal areas (where food is sold) and that animals are properly cared for and managed in public settings. In addition, this report recommends educating venue operators, staff, exhibitors, and visitors regarding the risk for disease transmission where animal contact is possible. PMID- 15788997 TI - Chemokine receptor gene expression in giant papillae of atopic keratoconjunctivitis. AB - PURPOSE: Major chemokine receptors in the giant papillae of atopic keratoconjunctivitis (AKC) have yet to be determined. We evaluated chemokine receptor genes and their ligand expressions in upper tarsal conjunctival giant papillae of AKC with atopic dermatitis and/or asthma. METHODS: CC, CXC, and CX3C chemokine receptor (R) gene expression levels in giant papillae of five clinically active AKC patients and in three age matched non-allergic control conjunctiva were measured with a multi-probe ribonuclease protection assay (RPA) system. The ligands of abundant chemokine receptors in the giant papillae were examined by immunohistochemistry or reverse transcription-polymerase chain reaction. Interleukin (IL)-4 and IL-13 gene expression levels were measured with RPA. Ligand expression in cultured human conjunctival fibroblasts was examined by reverse transcription-polymerase chain reaction. RESULTS: High CXCR4 and CCR4 gene expression levels were detected in the giant papillae of all (CXCR4) and four out of five (CCR4) patients. As a CCR4-ligand, thymus and activation regulated cytokine (TARC/CCL17) rather than macrophage derived chemokine (MDC/CCL22), was predominant immunohistochemicaly in the giant papillae. Giant papillae with high CCR4 gene expression levels showed high IL-4 and IL-13 expression. Cultured human conjunctival fibroblasts express stromal cell derived factor-1 (SDF-1/CXCL12) in vitro. CONCLUSIONS: CXCR4 and CCR4 are the major chemokine receptor genes expressed in the giant papillae of AKC with atopic dermatitis and/or asthma. Our findings suggest a role for CXCR4 and CCR4 in the formation of giant papillae. PMID- 15788998 TI - Over expression of FGF7 enhances cell proliferation but fails to cause pathology in corneal epithelium of Kerapr-rtTA/FGF7 bitransgenic mice. AB - PURPOSE: Available evidence suggests that fibroblast growth factor 7 (FGF7, also known as keratinocyte growth factor, KGF) serves as a paracrine growth factor modulating corneal epithelial cell proliferation. In the present study, we used a binary inducible transgenic mouse model to examine the role of FGF7 on corneal epithelium proliferation. METHODS: A keratocyte specific 3.2 kb murine keratocan promoter (Kerapr) was used to prepare Kerapr-rtTA transgenic (Kr) mice that constitutively overexpress reverse tetracycline transcription activator (rtTA) by cornea stromal keratocytes. The Kr mice were crossed with tet-O-FGF7 mice to produce Kr/tet-O-FGF7 bitransgenic mice. Expression of human FGF7 (hFGF7) was induced by the administration of doxycycline via intraperitoneal injection and/or feeding mice doxycycline in drinking water and chow. Overexpression of hFGF7 was confirmed by RT-PCR and western blot. BrdU incorporation was used to determine cell proliferation. RESULTS: The rtTA mRNA and protein were constitutively expressed by the cornea with or without doxycycline induction, whereas hFGF7 was detected only in Kr/tet-O-FGF7 bitransgenic mice upon induction by doxycycline. Examination of induction kinetics in adult Kr/tet-O-FGF7 bitransgenic mice after a single intraperitoneal injection of doxycycline revealed that hFGF7 mRNA expression was detected 12 h after doxycycline administration, peaked at 36 h, was sustained up to 48 h, and declined thereafter. The elevated level of hFGF7 expression coincided with hyperproliferation of corneal epithelial cells. In bitransgenic mice, the number of BrdU labeled cells increased after 36 and 48 h of transgene induction compared to controls of noninduced bitransgenic or doxycycline treated single transgenic mice. The BrdU labeling index was 33+/-9.2 positive cells per corneal section for Kr/tet-O-FGF7 bitransgenic mice and 25+/ 9.3 for tet-O-FGF7 single transgenic mice at 36 h post-doxycycline treatment. However, the excess FGF7 driven by doxycycline induction did not produce severe perturbation of corneal epithelium homeostasis. CONCLUSIONS: Our results demonstrate that the doxycycline inducible system is effective in regulating transgene expression in corneal stroma of Kr/tet-O-FGF7 bitransgenic mice. However, the development of pathology resulting from the overexpression of transgenes may depend on whether the amount of transgene product present is sufficient to alter the homeostasis of the targeted tissues. PMID- 15788999 TI - Bone morphogenetic proteins promote neurite outgrowth in retinal ganglion cells. AB - PURPOSE: The purpose of the present study is to test the ability of members of the transforming growth factor/bone morphogenetic protein family to influence retinal ganglion cell (RGC) survival and neurite outgrowth in primary cell culture using a high throughput analysis. METHODS: Primary cell cultures were generated using immunoselection of Thy-1 positive cells from dissociated postnatal rat retina and grown on poly-L-lysine/laminin coated 96 well culture dishes in the presence or absence of members of the transforming growth factor/bone morphogenetic protein family. High throughput analysis was performed following fluorescence staining with Hoechst, Calcein AM, and TOTO-3. Outcomes included overall cell survival, survival of cells with neurite outgrowth, and a variety of parameters of neurite outgrowth. RESULTS: Immunomagnetic selection led to an enrichment of cell cultures for RGCs (79%+/-6.8%). While no significant effect on overall survival was observed with any of the factors tested, members of the bone morphogenetic protein (BMPs) family (BMP2, BMP13, and GDF8 (growth differentiation factor 8)) and BDNF (brain derived neurotrophic factor) increased the number of surviving RGCs with neurite extension in a dose dependent manner. As a group, BMPs increased the number of neurites, length of neurites, and the number of branch points, while BDNF primarily increased neurite length and branch points. CONCLUSIONS: We have developed an efficient system that allows for high throughput analysis of cultures enriched for RGCs. Using this assay system, we found that BMPs promote the survival of outgrowth neurons and neurite development in RGC culture. PMID- 15789000 TI - Downregulation of the atrial natriuretic peptide/natriuretic peptide receptor-C system in the early stages of diabetic retinopathy in the rat. AB - PURPOSE: Atrial natriuretic peptide (ANP) is a known vascular antipermeability and antiangiogenic factor, but its possible alteration during the early stages of diabetic retinopathy has not yet been explored. The present study sought to investigate the expression of ANP and its receptors using a model of streptozotocin (STZ) induced diabetes in the rat. METHODS: Diabetes was induced in male Wistar rats by an intraperitoneal injection of STZ. Age matched animals served as control. One and 3 months after the onset of diabetes, the expression of ANP mRNA and that of its receptors (NPRA, NPRB, NPRC) and the immunoreactive ANP was quantified in retinal tissue by quantitative real time reverse transcription-polymerase chain reaction (RT-PCR) and radioimmunoassay, respectively. The locations of ANP and glial fibrillary acidic protein (GFAP) in normal and diabetic retinas were also established by immunohistochemistry. RESULTS: No alteration in the gene expression of the retinal natriuretic peptide system was noted after 1 month of diabetes. However, 3 months after the onset of diabetes, significantly diminished ANP and NPRC mRNA levels were detected in the retina of diabetic rats compared to controls, while NPRA, NPRB mRNA levels remained unchanged. At this time point, retinal ANP concentrations were significantly diminished in the diabetic rats compared to control rats. However, at 1 month retinal ANP concentrations in diabetic retina were similar to control rats. Diabetes caused the downregulation of ANP protein expression in the layers of the retina at 3 months after the induction of diabetes. ANP immunoreactivity was detected in the cell bodies of the astrocytes and in their processes enveloping vessels. CONCLUSIONS: The downregulation of ANP and NPRC in retinas of diabetic rats suggests a role for this peptide in experimental diabetic retinopathy. Further studies should address the possible involvement of the ANP/NPRC system in the pathophysiology of diabetic retinopathy. PMID- 15789001 TI - Bankart repair in patients aged fifty years or greater: results of arthroscopic and open repairs. AB - Currently, there is little information on the results of Bankart repairs in older patients. Therefore, the purpose of this study was to determine the results, complications, and rates of revision among patients aged 50 years or greater undergoing Bankart repairs. Between 1992 and 1999, 12 Bankart repairs were performed on patients aged 50 years or greater (mean, 57 years) at our institution. Eleven patients with complete clinical records, operative reports, and minimum 3-year follow-up (mean, 6.5 years) were included in the study. All patients had shoulder instability as a result of specific trauma. Six patients underwent open repairs, and five underwent arthroscopic repairs. There were no patients with full-thickness rotator cuff tears. At the most recent follow-up, there were no shoulders with recurrent instability. The mean simple shoulder test score was 10.8 in the open group and 9.8 in the arthroscopic group. The mean American Shoulder and Elbow Surgeons score was 98 in the open group versus 87 in the arthroscopic group. At the most recent follow-up, mean elevation in the open group was 178 degrees versus 174 degrees in the arthroscopic group. Mean external rotation was 70 degrees in the open group and 72 degrees in the arthroscopic group. The data from this study suggest that Bankart repair in older patients is associated with a low recurrence rate, and similar results may be obtained with either open or arthroscopic procedures. PMID- 15789002 TI - Shoulder arthroplasty for atraumatic avascular necrosis of the humeral head: nineteen shoulders followed up for a mean of seven years. AB - Nineteen shoulder arthroplasties for the treatment of nontraumatic avascular necrosis of the humeral head were evaluated. The osteonecrosis was idiopathic in 6 shoulders, was a result of corticotherapy in 10, occurred after radiation in 2, and occurred after Gaucher's disease in 1. A total shoulder arthroplasty was performed in 5 cases and a hemiarthroplasty in 14. At 7 years' follow-up (range, 2 to 12 years), there were 7 excellent, 9 satisfactory, and 3 unsatisfactory results. The Constant score averaged 58 points, for an adjusted score of 78%. Radiolucent lines were present around 2 glenoid components, and 1 was radiographically loose. In 2 cases with humeral head replacement, there was painful glenoid wear. Shoulder arthroplasty for nontraumatic avascular necrosis yields satisfactory results with a pain-free shoulder in more than 80% of cases. However, limitation of motion often persists. Better results can be expected with shorter preoperative delay, when preoperative pain is moderate and range of motion preserved and when the etiology is not postradiation avascular necrosis, which in our series yielded the worst results. PMID- 15789004 TI - The Constant score in normal shoulders. AB - Relative Constant-Murley shoulder scores are based on an age- and sex-matched normal population from Constant's original research. The purpose of this study was to determine normal Constant scores in a contemporary population and compare these values with those originally established by Constant. Two subject groups were analyzed. The first group comprised 1620 clinic patients whose normal shoulders were scored by resident physicians instructed on the Constant scoring technique. The second group comprised 115 healthy volunteers whose normal shoulders were scored by 1 experienced physician-researcher. There were no significant score differences between the two groups, indicating that, after instruction, scoring did not depend on tester experience. In both groups, there were significant score differences between sex and age groups. Similar to Constant's study, age-related declines in scores and strength existed for both sexes but were less pronounced. Using Constant's original values to calculate relative scores can overestimate shoulder function in women aged over 40 years and men aged over 60 years. If relative Constant scores are used, absolute scores should be concurrently reported to allow comparisons with different populations. PMID- 15789003 TI - Massive rotator cuff tears: the result of partial rotator cuff repair. AB - Massive rotator cuff tears often defy traditional repair techniques and have led to a variety of treatment recommendations. We reviewed retrospectively the results of partial repair of massive rotator cuff tears in 24 patients. Overall results were excellent in 11 patients (46%), good in 5 (21%), fair in 7 (29%), and poor in 1 (4%). Ninety-two percent of patients were satisfied with the result of surgery. Satisfactory pain relief was achieved in 83% (P<.001). Active elevation improved from 114 degrees to 154 degrees , and no patient lost strength after surgery. The ability to raise the arm to at least 135 degrees improved from 13 patients preoperatively to 21 patients postoperatively. This series serves as a basis of comparison to debridement, tendon transfers, and tendon augmentation procedures for the treatment of massive irreparable rotator cuff tears. PMID- 15789005 TI - Arthroscopic observation of capsulolabral reduction after shoulder dislocation. AB - The standard treatment for most first-time anterior shoulder dislocations is reduction and immobilization. This places the arm in a position of adduction and internal rotation. We question whether this position assists or hinders reduction of a displaced capsulolabral injury (Bankart lesion). This study examined the pathology present after traumatic first-time anterior shoulder dislocations and looked at the reduction of the Bankart lesion in internal and external rotation. A series of 25 patients with Bankart lesions were identified who underwent arthroscopy at a mean of 10.5 days after first-time anterior shoulder dislocations. We found that there is wide variation in pathology after anterior dislocation. In 23 of 25 patients in whom the capsulolabral complex was detached (Bankart lesion), we found that external rotation of the arm improved the reduction of the labrum (92%). We conclude that in patients with a displaced Bankart lesion, the standard treatment of immobilization in internal rotation may hinder anatomic healing of this injury. PMID- 15789006 TI - Biceps tenodesis associated with arthroscopic repair of rotator cuff tears. AB - Associated lesions of the biceps tendon are commonly found during arthroscopic repair of rotator cuff tears. These lesions are treated with tenodesis, classically performed through an open approach. However, it seems reasonable to seek a single approach to correct both lesions; therefore, we have proposed a new arthroscopic technique that allows an exclusive arthroscopic tenodesis by including the biceps tendon in the rotator cuff suture, a surgical technique with a single suture of the rotator cuff that includes the biceps tendon. We treated 97 shoulders in 96 patients arthroscopically for complete rotator cuff tears. Of these shoulders, 15 required tenodesis for treatment of biceps tendon lesions. Through an arthroscopic approach, a subacromial decompression followed by a rotator cuff repair was carried out in association with a biceps tenodesis. In this technique, one limb of the suture was passed through the biceps tendon, and the other was passed through the rotator cuff tear, bringing both tissues together in the final suture. Of the patients, 9 were men and 5 were women. Their mean age was 71 years (range, 41-80 years). The dominant arm was affected in all patients. Postoperative evaluation, by use of the UCLA score, after a mean follow up period of 32.4 months showed satisfactory results in 93.4% of patients: 11 had excellent results, 3 had good results, and only 1 had an unsatisfactory result. In this case a postoperative magnetic resonance imaging scan showed an intact rotator cuff and biceps tenodesis. The suture involving the rotator cuff and the biceps tendon proved effective to correct both lesions, with the main advantage being that an additional approach was not required. PMID- 15789007 TI - Accuracy of subacromial injection: anterolateral versus posterior approach. AB - Subacromial bursa injection is commonly performed via an anterolateral or posterior approach. No study has compared the accuracy rates of these approaches. Twenty cadaveric shoulders were injected with radiocontrast in the subacromial bursa via an anterolateral approach and twenty via a posterior approach. Ten shoulders were injected with methylene blue via each technique for dissection. The anterolateral approach was accurate by fluoroscopy in 18 shoulders (90%), but dissection of 10 shoulders revealed successful injection in only 6 (60%). The posterior approach was judged to be successful in 16 shoulders (80%), and dissection of 10 shoulders confirmed 8 injections (80%) were accurate. There was no significant difference in the accuracy by fluoroscopy (P=.38) or dissection (P=.33). The accuracy rates of the anterolateral and posterior approaches to subacromial bursa injections are not significantly different. The anterolateral approach may place injected material medial to the medial bursal boundary, and fluoroscopy may not accurately assess this placement. PMID- 15789008 TI - Excessive radiofrequency application: effects on capsular tissue in an animal model. AB - Capsular attenuation has been seen after thermal capsulorrhaphy surgery. The purpose of this study was to evaluate the mechanical, histologic, and morphologic effects on capsular tissue after cumulative applications of radiofrequency energy. Ovine patellofemoral capsular tissue was treated with 1, 2, 4, or 8 applications of bipolar radiofrequency energy and then analyzed. No acute capsular ablation or destruction was seen grossly, even in the 8-application group. No definitive visual clues that excessive radiofrequency energy had been applied were seen. There was significant shrinkage and loss of tensile stiffness for all thermal application groups. Given the small sample sizes, post application failure load, percent relaxation, and stiffness were not observed to be significantly different among the groups. Cumulative applications produced minimal further tissue shrinkage but were accompanied by larger, though not statistically significant, mechanical property losses and increased depth of tissue penetration. These findings suggest that there is no benefit to repeated applications of radiofrequency energy to capsular tissue. PMID- 15789009 TI - Open posterior stabilization for recurrent posterior glenohumeral instability. AB - Open posterior capsular shift is used for posterior glenohumeral instability that has failed nonoperative treatment. Few series have fully evaluated the outcome after open posterior stabilization. The purpose of this series was to evaluate the clinical and radiographic outcome after open posterior stabilization of the shoulder. Preoperative and intraoperative factors were analyzed with regard to their impact on results. Forty-eight consecutive shoulders were identified that had undergone primary open shoulder stabilization by use of open posterior capsular shift. Of the shoulders, 4 were lost to follow-up, resulting in a study group of 44 shoulders in 41 patients. Shoulders were evaluated at a range of 1.8 to 22.5 years after surgery by use of the L'Insalata shoulder form, Short Form-36 (SF-36), and a subjective shoulder rating in 44 shoulders. Thirty-nine shoulders were evaluated by physical examination, and thirty-seven underwent radiographic examination. A recurrence of posterior instability occurred in 8 shoulders (19%). Of the patients, 84% were satisfied with the current status of their shoulder. The mean L'Insalata score was 81.25+/-17.8 points, the mean SF-36 physical component score was 50.81+/-7.87, and the mean mental component score was 53.82+/ 7.55. Significantly poorer satisfaction and outcome scores were seen in shoulders found to have a chondral defect at the time of stabilization and in patients aged greater than 37 years at the time of surgery. No progressive radiographic signs of glenohumeral arthritis were seen up to 22 years after surgery. Open posterior shoulder stabilization is a reliable procedure for treating significant posterior instability without causing arthritic changes. Patients found to have chondral damage within the shoulder and older patients were found to have less success after stabilization. PMID- 15789010 TI - Differential patterns of muscle activation in patients with symptomatic and asymptomatic rotator cuff tears. AB - Patients with rotator cuff tears have varying degrees of symptom expression. Our purpose was to evaluate the differential firing patterns of the rotator cuff, deltoid, and scapular stabilizer muscle groups in normal control subjects and in patients with symptomatic and asymptomatic 2-tendon rotator cuff tears. Eighteen subjects were evaluated: six normal subjects and twelve with 2-tendon cuff tears (six asymptomatic and six symptomatic). All cuff tear patients had magnetic resonance imaging (MRI) scans documenting superoposterior tear configurations involving the supraspinatus and infraspinatus tendons; all normal subjects had an ultrasound examination confirming the absence of cuff pathology. Subjects were grouped based on shoulder examination and outcomes questionnaires. Asymptomatic patients had minimal pain (<3 on the visual analog scale and no loss of active range of motion compared with the contralateral side); symptomatic patients had pain greater than 3 on the visual analog scale and decreased range of motion compared with the contralateral side (>10 degrees of motion loss). Electromyographic activity from 12 muscles and kinematic data were collected simultaneously during 10 functional tasks. Both symptomatic and asymptomatic cuff subjects demonstrated a trend toward increased muscle activation during all tasks compared with normal subjects. During the internal rotation tasks, asymptomatic patients had significantly greater (P<.05) subscapularis activity than symptomatic patients (65% maximal voluntary contraction [MVC] vs 42% MVC). During the carrying task, asymptomatic patients demonstrated significantly less (P<.03) upper trapezius muscle activation than symptomatic patients (16% MVC vs 50% MVC). During shoulder elevation tasks, symptomatic patients had significantly greater supraspinatus (52% MVC vs 28% MVC, P<.03), infraspinatus (32% MVC vs 16% MVC, P<.05), and upper trapezius (39% MVC vs 20% MVC, P<.04) muscle activation compared with asymptomatic patients. During heavy elevation (8 lb), asymptomatic patients showed a trend toward increased activation (P<.06) of the subscapularis compared with symptomatic patients (34% MVC vs 21% MVC). Differential shoulder muscle firing patterns in patients with rotator cuff pathology may play a role in the presence or absence of symptoms. Asymptomatic subjects demonstrated increased firing of the intact subscapularis, whereas symptomatic subjects continued to rely on torn rotator cuff tendons and periscapular muscle substitution, resulting in compromised function. PMID- 15789011 TI - Magnetic resonance imaging findings of refractory tennis elbows and their relationship to surgical treatment. AB - Treatment of tennis elbow is a problem. We performed magnetic resonance imaging (MRI) examinations of 11 elbows in 11 patients who were referred to us with refractory tennis elbow. Of the 11 elbows, 6 showed a high signal intensity focus on MRI T2 images in the tendon of the extensor carpi radialis brevis at the lateral epicondyle. The other 5 elbows showed various abnormal findings on MRI. The mean age of the 6 patients with a high T2 signal focus was 52.0 years, and the mean morbidity period was 25 months. We performed enucleation of the granulation focus through a surgical approach without detachment of the tendon origin of the extensor carpi radialis brevis. Pain was relieved and elbow function recovered in all 6 cases. This study demonstrates that MRI is an important decision-making tool in the surgical treatment of this condition. Enucleation of the granulation focus with high T2 signal has proved to be effective in 6 selected cases. PMID- 15789013 TI - Geometrical analysis of Copeland surface replacement shoulder arthroplasty in relation to normal anatomy. AB - This study examines the effect of humeral head surface replacement on the geometry of the arthritic glenohumeral joint and correlates changes to clinical outcome. Thirty-nine patients undergoing resurfacing arthroplasty were prospectively followed up clinically and radiologically for a mean of 38 months (range, 24 to 72 months). Measurements were taken from coronal radiographs by use of a validated technique. There was an estimated mean 6 mm erosion of preoperative lateral offset (coracoid base to greater tuberosity). The prosthesis reduced the humeral head radius by a mean of 3.5%. Humeral offset (greater tuberosity to center of instant rotation) increased by a mean of 5 mm (95% CI, 3.8 to 6.4; P<.01). Mean age-/sex-adjusted Constant scores improved from 26 to 83 points (P<.01). The improved postoperative motion showed a nonlinear relationship with humeral offset. We conclude that surface replacement restores humeral offset and the lever arm of the deltoid and supraspinatus while otherwise mimicking individual anatomy. Function depends on adequate restoration of this lever arm without overstuffing the joint. PMID- 15789012 TI - Operative treatment of displaced medial epicondyle fractures in children and adolescents. AB - Twenty-five patients who sustained displaced medial epicondyle fractures were treated by various surgical techniques. There were 18 males and 7 females. The mean age was 13.7 years. Surgical outcome was evaluated by use of the Elbow Assessment Score of the Japanese Orthopedic Association. Many variables that might influence the surgical outcome were considered. The results revealed no significant correlation between surgical outcome and injury mechanism, displacement, interval from injury to surgery, dislocation, fixation method, or duration of immobilization. The motion arc was moderately decreased in 1 patient treated by use of screw fixation. Moderate instability to valgus stress was noted in another patient treated by use of pin fixation (K-wires). The treatment results were all scored, and the patients with medial epicondylar fractures (displacement >5 mm) showed good to excellent results with operative treatment. Residual deformity did not compromise cosmetic appearance or clinical results. Therefore, operative treatment is a suitable choice for managing these fractures in children and adolescents. PMID- 15789014 TI - Tensile strength of ligaments after thermal shrinkage depending on time and immobilization: in vivo study in the rabbit. AB - The effects of immobilization on healing collagenous tissue treated with thermal shrinkage are investigated in an in vivo rabbit model. Thermal shrinkage was performed on 45 of 50 medial collateral ligaments in 25 mature New Zealand rabbit knees. In half of the knees, the joints were immobilized. Animals were killed at 0, 3, 6, and 9 weeks postoperatively. Failure loads of ligaments were determined, and morphologic changes were evaluated by electron microscopy. The failure load of shrunken ligaments was significantly lower than that of the intact ligaments on the day of operation (P<.05). Shrunken ligaments reached their highest failure loads in the third week within the first 9 weeks. The immobilized ligaments remained weaker than the mobile group, but this difference was statistically significant only in the ninth week (P<.05). Immobilization seems to have negative effects on the healing ligament. Ideal timing for remobilization is still controversial, and abandoning immobilization protocols for longer than 3 weeks should be considered. PMID- 15789015 TI - Structural and mechanical properties of the glenohumeral joint posterior capsule. AB - The purpose of this study was to quantify regional variations in material properties of the glenohumeral joint posterior capsule and to compare these data with the anterior band of the inferior glenohumeral ligament (AB-IGHL). Mechanical properties were determined for individual bands of the AB-IGHL, superior posterior capsule (SUP-PC), middle posterior capsule (MID-PC), and inferior posterior capsule (INF-PC). Significant differences in tissue thickness were found among the 3 posterior capsular regions and the AB-IGHL. The AB-IGHL was thicker than the MID-PC (P=.03) and INF-PC (P=.01), and the SUP-PC was thicker than the INF-PC (P=.02). Except for significant differences in failure strains, material properties were not significantly different among the 4 tissue regions. There were no significant differences between tissue bands in modulus (P=.2), maximum stress (P=.46), or strain energy density (P=.62). Specimens failed primarily near the glenoid insertion (75%), with 4 specimens failing at the humeral insertion and 2 others failing in the tissue's mid substance. PMID- 15789016 TI - A biomechanical comparison of four reconstruction techniques for the medial collateral ligament-deficient elbow. AB - The initial strength of the intact medial collateral ligament (MCL) of the elbow and the strength of 4 reconstruction techniques were compared. Twenty cadaveric upper extremities were mounted in a custom jig with the elbow at 90 degrees , and a pneumatic cyclic valgus loading protocol was used. The mean peak load to failure was 142.5+/-39.4 N for the intact ligaments and 53.0+/-9.5 N for the docking reconstructions, 52.5+/-10.4 N for the EndoButton reconstructions, 41.0+/ 16.0 N for the interference screw reconstructions, and 33.3+/-7.1 N for the figure-eight reconstructions. The peak load to failure of the MCL reconstructions was inferior compared with the intact ligament (P<.001). No difference in strength was found between the docking and single-strand medial collateral reconstruction with the use of an EndoButton for ulnar fixation (P>.05, beta=.14). Both of these reconstruction methods were stronger than the interference screw or figure-eight technique (P<.004). The optimal fixation method for a single-strand MCL reconstruction may require improved interference screws or a modified EndoButton procedure. PMID- 15789017 TI - Transient brachial plexus block after shoulder manipulation: a case report. PMID- 15789018 TI - Acute subscapularis tendon rupture caused by throwing: a case report. PMID- 15789019 TI - Displaced periprosthetic humeral fracture treated with functional bracing: a report of two cases. PMID- 15789020 TI - A case of traumatic divergent fracture-dislocation of the elbow combined with Essex-Lopresti lesion in an adult. PMID- 15789021 TI - Idiopathic Staphylococcus aureus necrotizing fasciitis of the upper extremity. PMID- 15789023 TI - To the editor. PMID- 15789024 TI - Unusual neonatal presentation of incontinentia pigmenti with persistent pulmonary hypertension of the newborn: a case report. AB - Incontinentia pigmenti (Bloch-Sulzberger syndrome) is a multisystem disorder with classical changing skin lesions. The other systems that are involved include the central nervous system, eye, hair, teeth, musculoskeletal system and, occasionally, the cardiovascular system. We report a neonate with a diagnosis of incontinentia pigmenti who presented at birth with pulmonary hypertension. This presentation has not been described in the literature. PMID- 15789025 TI - SIDS among twins: a confounded relationship. PMID- 15789029 TI - Strategies for the reconstitution of chromatin. AB - In eukaryotes, chromatin is the natural form of DNA in the nucleus. For hundreds of millions of years, DNA-binding factors have evolved with chromatin. It is therefore more desirable to study the molecular mechanisms of DNA-directed processes with chromatin than with naked DNA templates. To this end, it is necessary to reconstitute DNA and histones into chromatin. Fortunately, there are a variety of methods by which a nonspecialist can prepare chromatin of high quality. Here, we describe strategies and techniques for the reconstitution of chromatin in vitro. PMID- 15789030 TI - Large-scale database searching using tandem mass spectra: looking up the answer in the back of the book. AB - Database searching is an essential element of large-scale proteomics. Because these methods are widely used, it is important to understand the rationale of the algorithms. Most algorithms are based on concepts first developed in SEQUEST and PeptideSearch. Four basic approaches are used to determine a match between a spectrum and sequence: descriptive, interpretative, stochastic and probability based matching. We review the basic concepts used by most search algorithms, the computational modeling of peptide identification and current challenges and limitations of this approach for protein identification. PMID- 15789031 TI - Kinomics: methods for deciphering the kinome. AB - Phosphorylation by protein kinases is the most widespread and well-studied signaling mechanism in eukaryotic cells. Phosphorylation can regulate almost every property of a protein and is involved in all fundamental cellular processes. Cataloging and understanding protein phosphorylation is no easy task: many kinases may be expressed in a cell, and one-third of all intracellular proteins may be phosphorylated, representing as many as 20,000 distinct phosphoprotein states. Defining the kinase complement of the human genome, the kinome, has provided an excellent starting point for understanding the scale of the problem. The kinome consists of 518 kinases, and every active protein kinase phosphorylates a distinct set of substrates in a regulated manner. Deciphering the complex network of phosphorylation-based signaling is necessary for a thorough and therapeutically applicable understanding of the functioning of a cell in physiological and pathological states. We review contemporary techniques for identifying physiological substrates of the protein kinases and studying phosphorylation in living cells. PMID- 15789032 TI - Biphasic cytochrome c release after transient global ischemia and its inhibition by hypothermia. AB - Hypothermia is effective in preventing ischemic damage. A caspase-dependent apoptotic pathway is involved in ischemic damage, but how hypothermia inhibits this pathway after global cerebral ischemia has not been well explored. It was determined whether hypothermia protects the brain by altering cytochrome c release and caspase activity. Cerebral ischemia was produced by two-vessel occlusion plus hypotension for 10 mins. Body temperature in hypothermic animals was reduced to 33 degrees C before ischemia onset and maintained for 3 h after reperfusion. Western blots of subcellular fractions revealed biphasic cytosolic cytochrome c release, with an initial peak at about 5 h after ischemia, which decreased at 12 to 24 h, and a second, larger peak at 48 h. Caspase-3 and -9 activity increased at 12 and 24 h. A caspase inhibitor, Z-DEVD-FMK, administered 5 and 24 h after ischemia onset, protected hippocampal CA1 neurons from injury and blocked the second cytochrome c peak, suggesting that caspases mediate this second phase. Hypothermia (33 degrees C), which prevented CA1 injury, did not inhibit cytochrome c release at 5 h, but reduced cytochrome c release at 48 h. Caspase-3 and -9 activity was markedly attenuated by hypothermia at 12 and 24 h. Thus, biphasic cytochrome c release occurs after transient global ischemia and mild hypothermia protects against ischemic damage by blocking the second phase of cytochrome c release, possibly by blocking caspase activity. PMID- 15789033 TI - Cerebral blood flow and brain oxygenation in rats breathing oxygen under pressure. AB - Hyperbaric oxygen (HBO(2)) increases oxygen tension (PO(2)) in blood but reduces blood flow by means of O(2)-induced vasoconstriction. Here we report the first quantitative evaluation of these opposing effects on tissue PO(2) in brain, using anesthetized rats exposed to HBO(2) at 2 to 6 atmospheres absolute (ATA). We assessed the contribution of regional cerebral blood flow (rCBF) to brain PO(2) as inspired PO(2) (PiO(2)) exceeds 1 ATA. We measured rCBF and local PO(2) simultaneously in striatum using collocated platinum electrodes. Cerebral blood flow was computed from H(2) clearance curves in vivo and PO(2) from electrodes calibrated in vitro, before and after insertion. Arterial PCO(2) was controlled, and body temperature, blood pressure, and EEG were monitored. Scatter plots of rCBF versus PO(2) were nonlinear (R(2)=0.75) for rats breathing room air but nearly linear (R(2)=0.88-0.91) for O(2) at 2 to 6 ATA. The contribution of rCBF to brain PO(2) was estimated at constant inspired PO(2), by increasing rCBF with acetazolamide (AZA) or decreasing it with N-nitro-L-arginine methyl ester (L NAME). At basal rCBF (78 mL/100 g min), local PO(2) increased 7- to 33-fold at 2 to 6 ATA, compared with room air. A doubling of rCBF increased striatal PO(2) not quite two-fold in rats breathing room air but 13- to 64-fold in those breathing HBO(2) at 2 to 6 ATA. These findings support our hypothesis that HBO(2) increases PO(2) in brain in direct proportion to rCBF. PMID- 15789036 TI - Cellular and molecular signal transduction pathways modulated by rituximab (rituxan, anti-CD20 mAb) in non-Hodgkin's lymphoma: implications in chemosensitization and therapeutic intervention. AB - The clinical application of rituximab (chimeric mouse anti-human CD20 mAb, Rituxan, IDEC-C2B8), alone and/or combined with chemotherapy, has significantly ameliorated the treatment outcome of patients with relapsed and refractory low grade or follicular non-Hodgkin's lymphoma (NHL). The exact in vivo mechanisms of action of rituximab are not fully understood, although antibody-dependent cell mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and apoptosis have been suggested. We have proposed that modifications of the cellular signaling pathways by rituximab may be crucial for its clinical response. The B-cell restricted cell surface phosphoprotein CD20 is involved in many cellular signaling events including proliferation, activation, differentiation, and apoptosis upon crosslinking. Monomeric rituximab chemosensitizes drug-resistant NHL cells via selective downregulation of antiapoptotic factors through the type II mitochondrial apoptotic pathway. Several signaling pathways are affected by rituximab which are implicated in the underlying molecular mechanisms of chemosensitization. ARL (acquired immunodeficiency syndrome (AIDS)-related lymphoma) and non-ARL cell lines have been examined as in vitro model systems. In ARL, rituximab diminishes the activity of the p38MAPK signaling pathway resulting in inhibition of the interleukin (IL)-10/IL-10R autocrine/paracrine cytokine autoregulatory loop leading to the inhibition of constitutive STAT-3 activity and subsequent downregulation of Bcl-2 expression leading to chemosensitization. Rituximab upregulates Raf-1 kinase inhibitor protein (RKIP) expression in non-ARL cells. Through physical association with Raf-1 and nuclear factor kappaB (NF-kappa B) inducing kinase (NIK), RKIP negatively regulates two major survival pathways, namely, the extracellular signal-regulated kinase1/2 (ERK1/2) and the NF-kappa B pathways, respectively. Downmodulation of the ERK1/2 and NF-kappa B pathways inhibits the transcriptional activity of AP-1 and NF-kappa B transcription factors, respectively, both of which lead to the downregulation of Bcl-(xL) (Bcl 2 related gene (long alternatively spliced variant of Bcl-x gene)) transcription and expression and sensitization to drug-induced apoptosis. Bcl-(xL) overexpressing cells corroborated the pivotal role of Bcl-(xL) in chemosensitization. The specificity of rituximab-mediated signaling and functional effects were corroborated by the use of specific pharmacological inhibitors. Many patients do not respond and/or relapse and the mechanisms of unresponsiveness are unknown. Rituximab-resistant B-NHL clones were generated to investigate the acquired resistance to rituximab-mediated signaling, and chemosensitization. Resistant clones display different phenotypic, genetic and functional properties compared to wild-type cells. This review summarizes the data highlighting a novel role of rituximab as a signal-inducing antibody and as a chemosensitizing agent through negative regulation of major survival pathways. Studies presented herein also reveal several intracellular targets modified by rituximab, which can be exploited for therapeutic and prognostic purposes in the treatment of patients with rituximab- and drug-refractory NHL. PMID- 15789037 TI - Outcomes, what outcomes? PMID- 15789034 TI - Bilirubin production and oxidation in CSF of patients with cerebral vasospasm after subarachnoid hemorrhage. AB - Delayed cerebral vasospasm after subarachnoid hemorrhage (SAH) remains a significant cause of mortality and morbidity; however, the etiology is, as yet, unknown, despite intensive research efforts. Research in this laboratory indicates that bilirubin and oxidative stress may be responsible by leading to formation of bilirubin oxidation products (BOXes), so we investigated changes in bilirubin concentration and oxidative stress in vitro, and in cerebral spinal fluid (CSF) from SAH patients. Non-SAH CSF, a source of heme oxygenase I (HO-1), and blood were incubated, and in vitro bilirubin production measured. Cerebrospinal fluid from SAH patients was collected, categorized using stimulation of vascular smooth muscle metabolism in vitro, and information obtained regarding occurrence of vasospasm in the patients. Cerebral spinal fluid was analyzed for hemoglobin, total protein and bilirubin, BOXes, malonyldialdehyde and peroxidized lipids (indicators of an oxidizing environment), and HO-1 concentration. The formation of bilirubin in vitro requires that CSF is present, as well as whole, non-anti-coagulated blood. Bilirubin, BOXes, HO-1, and peroxidized lipid content were significantly higher in CSF from SAH patients with vasospasm, compared with nonvasospasm SAH CSF, and correlated with occurrence of vasospasm. We conclude that vasospasm may be more likely in patients with elevated BOXes. The conditions necessary for the formation of BOXes are indeed present in CSF from SAH patients with vasospasm, but not CSF from SAH patients without vasospasm. PMID- 15789038 TI - Risk-based recall intervals recommended. AB - This summary of clinical guideline 19 from NICE includes recommendations for patients of all ages (both dentate and edentulous patients) and covers primary care received from NHS dental staff (dentists, independent contractors contracting within the NHS, dental hygienists and therapists) practising in England and Wales. The guideline takes into account the potential of the patient and the dental team to improve or maintain the quality of life and to reduce morbidity associated with oral and dental disease. The guideline does not cover intervals between dental examinations that are not routine dental recalls; that is, intervals between examinations related to ongoing courses of treatment, or part of current dental interventions, nor does it cover emergency dental interventions, or intervals between episodes of specialist care. PMID- 15789039 TI - Insufficient evidence to understand effect of routine scaling and polishing. AB - DATA SOURCES: Relevant studies were sourced using the Cochrane Oral Health Group Trials Register, the Cochrane Central Register of Controlled Trials, Medline and Embase. Reference lists from relevant articles were scanned and the authors of eligible studies were contacted where possible to identify trials and obtain additional information. STUDY SELECTION: Trials were selected if they were randomised, included anyone with an erupted permanent dentition, and where subjects were judged to have received a routine scale and polish as defined in this review. Outcomes assessed included tooth loss, plaque, calculus, gingivitis, bleeding and periodontal indices, changes in probing depth, attachment change, patient-centred outcomes and economic outcomes. DATA EXTRACTION AND SYNTHESIS: Trial details were independently extracted, in duplicate, by two reviewers. Authors were contacted where possible and where deemed necessary for further details regarding study design and for data clarification. A quality assessment of all included trials was carried out. The Cochrane Collaboration's statistical guidelines were followed and both the standardised mean differences and weighted mean differences were calculated, as appropriate, using random-effects models. RESULTS: Eight studies were included in this review and all studies were assessed as having a high risk of bias. Two split-mouth studies provided data for the comparison between scale and polish versus no scale and polish. One study, which involved people attending a recall programme following periodontal treatment, found no statistically significant differences for plaque, gingivitis and attachment loss between experimental and control units at each timepoint during the 1-year trial. The other study, of adolescents in a developing country who had high existing levels of and calculus who had not received any dental treatment for at least 5 years, reported statistically significant improvements in calculus and gingivitis (bleeding) scores between treatment and control units at 6, 12 and 22 months following a single scale and polish provided at baseline to treatment units. For comparisons between routine scale and polish procedures provided at different time intervals, there were some statistically significant differences in favour of scaling and polishing carried out at more frequent intervals, that is, at 2 weeks versus 6 months; 2 weeks versus 12 months (for the outcomes plaque, gingivitis, pocket depth and attachment change); and at 3 months versus 12 months (for the outcomes plaque, calculus and gingivitis). There were no studies comparing the effects of scaling and polishing provided by dentists or professionals complementary to dentistry. CONCLUSIONS: The research evidence is not of sufficient quality to reach any conclusions regarding the beneficial and adverse effects of routine scaling and polishing for periodontal health and regarding the effects of providing this intervention at different time intervals. High-quality clinical trials are required to address the basic questions posed in this review. Scaling and/or polishing of the crown and root surfaces of the teeth to remove local irritational factors (plaque, calculus, debris and staining), that does not involve periodontal surgery or any form of adjunctive periodontal therapy such as the use of chemotherapeutic agents or root planing. PMID- 15789040 TI - Dietary factors and dental caries. AB - DATA SOURCES: Articles were sourced using Medline, the Cochrane Library, reference lists of identified articles and selected textbooks. STUDY SELECTION: Studies chosen for inclusion in the review were randomised or controlled clinical trials of at least 2 years' duration that used caries increment in the permanent or primary dentition as the end point. Publications in Danish, English, French, German, Italian, Norwegian, Spanish or Swedish were included. For multiply reported trials the one with the longest follow-up period was included. DATA EXTRACTION AND SYNTHESIS: Inclusion decisions and grading of the studies was carried out independently by two of the authors. The main outcome was caries increment and the measure of treatment effect was either relative risk reduction or prevented faction. A qualitative synthesis of the included studies was conducted. RESULTS: Eighteen studies met the inclusion criteria. They included the total or partial substitution of sucrose with sugar substitutes or the addition of protective foods to chewing gum. No study could be found that had evaluated the effect of information designed to reduce sugar intake/frequency as a single preventive measure. It is suggested that the evidence for the use of sorbitol or xylitol in chewing gum, or for the use of invert sugar, is inconclusive. No caries-preventive effect was found from adding calcium phosphate or dicalcium phosphate dihydrate to chewing gums. CONCLUSIONS: The review dearly demonstrates the need for well-designed randomised clinical studies, with adequate control groups and high compliance, looking at the effect of dietary measures on dental caries. PMID- 15789041 TI - Atraumatic restorative techniques could reduce discomfort in children receiving dental treatment. AB - DESIGN: This was a randomised controlled trial conducted in Indonesia. INTERVENTION: Children, who each had at least one multisurface cavity in a deciduous molar, were randomly allocated to two treatment groups. The test group was treated using atraumatic restorative techniques (ART) using only hand instruments. The control group received treatment using rotary instruments (operated at 750 rpm), without water cooling. Glass ionomer cement was used for restoration in both groups. OUTCOME MEASURE: The extent of discomfort was assessed by measuring the behavioural (psychological) and the physiological aspect represented by, respectively, a modified Venham score and the heart rate of the children at six fixed moments during dental treatment: (i) when the child entered the treatment room, (ii) at the start of excavation, (iii) at the moment of deepest excavation, (iv) at the moment of application of the matrix band and wedges, (v) at the moment the restoration was applied, and (vi) after completion of the treatment. RESULTS: A total of 403 children were treated. Venham scores showed a marked difference between the two groups at most timepoints. Heart rate measurements were different at deep excavation. Also, a clear relation between Venham scores and heart rate measurements could be found at all time points. Confounding could be shown for operating dentist, gender of the patient and initial anxiety, but not for age. No effect modification could be shown. CONCLUSIONS: Children treated according to the ART approach using hand instruments alone experience less discomfort than those treated using rotary instruments. PMID- 15789042 TI - No benefit from prophylactic antibiotics in third molar surgery. AB - DESIGN: This was a randomised controlled trial in a hospital environment. INTERVENTION: After surgical removal of third molars postoperative treatment was with oral amoxicillin-clavulanic acid (AC) or clindamycin (CL) or no antibiotics (C). The surgical technique was the same in all cases, and the follow-up period was 4 weeks. OUTCOME MEASURE: Parameters evaluated were pain, differences in mouth opening, infection, the occurrence of dry socket, and adverse postoperative side effects. RESULTS: A total of 528 lower third molars were surgically removed in 288 patients during a period of 30 months. The patients' mean age was 20.7 years (age range, 14-61 years). No severe complications such as perimandibular abscess or cellulitis occurred in any patient in any group. There was no significant difference between the groups in the overall occurrence of local infection symptoms after surgery (range, 3.4-4.4%; mean, 3.98%), nor for other parameters. Interestingly, 69.6% of the patients with dry sockets had partially erupted third molars. This rate was the same in each group (62.5% versus 75%) and did not vary significantly. Reported adverse effects were similar in each group (15.3% for AC, 12.2% for CL, 13.9% for C). CONCLUSIONS: The results show that specific postoperative oral prophylactic antibiotic treatment after the removal of lower third molars does not contribute to better wound healing, less pain nor increased mouth opening, and could not prevent the cases of inflammatory problems after surgery, respectively. It is therefore not recommended for routine use. PMID- 15789043 TI - Dental procedures can be undertaken without alteration of oral anticoagulant regimen. AB - DATA SOURCES: Medline provided the primary data source with references from identified articles being reviewed for additional studies. The Cochrane Collaboration database was also searched and a search performed of cited references. STUDY SELECTION: Clinical studies, in English, examining perioperative management of patients receiving long-term oral anticoagulant (OAC) therapy were selected. DATA EXTRACTION AND SYNTHESIS: Data were extracted regarding management strategy, thrombo-embolic events and bleeding complications, and type of surgical or invasive procedure. Event rates were reported as number of patients experiencing the event divided by number of patients at risk. Binomial and Poisson distributions were used to calculate 95% confidence intervals (CI). RESULTS: A total of 31 reports were identified and concluded to be of generally poor quality. For studies reporting thrombo-embolic events, 29 events occurred in 1868 patients (1.6%; 95% CI, 1.0-2.1) of which seven were strokes (0.4%; 95% CI, 0.0-0.7). Major bleeding while receiving OAC was reported to be rare for dental procedures (occurring in four out of 2014 individuals), arthrocentesis (in none out of 32), cataract surgery (none out of 203), and upper endoscopy or colonoscopy with or without biopsy (no occurrences in 111 patients). For the other invasive and surgical procedures reviewed, OAC needs to be withheld and a suitable personalised perioperative management strategy instigated. A guideline, based on the limited evidence available, for the perioperative management of anticoagulation for procedures requiring discontinuation of OAC is presented. CONCLUSIONS: Certain surgical or invasive procedures can be undertaken in patients who are taking OAC therapy without alteration of their regimen. For procedures requiring discontinuation of OAC, personalised management strategies are required. More rigorous studies are needed to better inform this debate. PMID- 15789044 TI - Leukoplakia prevalence estimate lower than expected. AB - DATA SOURCES: Data were sourced using Medline and Embase. STUDY SELECTION: Initial selection criteria were that study samples should be representative of the underlying adult population and that the leukoplakia diagnostic criteria used by the examiners should have been widely accepted ones. Because most published studies did not comply with these requisites, modified criteria were as follows. First, the sample, even if not representative of the underlying study population, should not have been collected from surgical departments of dental hospitals or clinics, where leukoplakia is usually treated (these studies would overestimate the disease prevalence). Secondly, even if the examiners were not calibrated, the diagnostic criteria used should be consistent with the internationally accepted criteria and must have been reported in the text. Thirdly, to make statistical analysis possible even if leukoplakia prevalence was not reported as a proportion, the overall number of sampled subjects and the number with the condition must be easy to extrapolate with a high level of accuracy. DATA EXTRACTION AND SYNTHESIS: Data were pooled using inverse variance weighting and random-effect methods. A sensitivity analysis was performed. RESULTS: A total of 23 primary studies was included, giving point-prevalence estimates with the inverse variance method of 1.49% [95% confidence interval (CI), 1.42-1.56] and 2.60% (95% CI, 1.72-2.74) with the random-effects method. The high between-study heterogeneity and the sensitivity analyses suggested that the second estimate was more reliable. Leukoplakia was significantly more prevalent in males (prevalence ratio, 3.22), but no difference was found between geographical areas and between younger and older adults. Using these data, the crude annual oral cancer incidence rate attributable to leukoplakia would be between 6.2 and 29.1 cases per 100 000 people. CONCLUSIONS: The pooled leukoplakia prevalence estimate was lower than expected. The estimated oral cancer incidence rate due to leukoplakia malignant transformation was high, however, and suggested that the global number of oral cancer cases is probably under-reported, particularly in developing countries. PMID- 15789045 TI - Although leukoplakia responds to some treatments relapses and adverse effects are common. AB - DATA SOURCES: The Cochrane Oral Health Group's Trials Register, Cochrane Central Trials Register, Medline and Embase were searched, as well as the following journals: Cancer, Community Dentistry and Oral Epidemiology, European Journal of Oral Sciences, Journal of Dental Research, Oral Oncology, Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontics. The reference lists of included studies and reviews were checked, and oral medicine experts were contacted through a European mailing list (EURORALMED). STUDY SELECTION: Randomised controlled trials (RCT) were included if they had enrolled patients who had a diagnosis of oral leukoplakia and who were undergoing any surgical or medical (topical and systemic) treatment. The primary outcome considered was malignant transformation of leukoplakia demonstrated by histopathological examination. Other outcomes considered were clinical resolution, histological modification and frequency of adverse effects. DATA EXTRACTION AND SYNTHESIS: Data were collected using a specific extraction form. The validity of studies included was assessed by two reviewers, on the basis of the method of allocation concealment, blindness of the study and loss of participants. Data were analysed by calculating relative risk (RR). When valid and relevant data were collected, a meta-analysis of the data was undertaken. RESULTS: The possible effectiveness of surgical interventions, including laser therapy and cryotherapy, has never been studied by means of an RCT. Nineteen RCT of nonsurgical interventions were identified: seven were included. Vitamin A and retinoids were tested in five RCT (245 patients); the other drugs tested were bleomycin (one study), mixed tea (one study) and beta-carotene (one study). Malignant transformation was recorded in just two studies. None of the treatments tested showed a benefit compared with placebo. Treatment with beta-carotene and vitamin A or retinoids was associated with significant rates of clinical resolution, compared with placebo or absence of treatment. Whenever reported, a high rate of relapse was a common finding. Side effects of variable severity were often described but interventions were well accepted by patients, since dropout rates were similar between treatment and control. CONCLUSIONS: To date there is no evidence of effective treatment in preventing malignant transformation of leukoplakia. Treatments may be effective in the resolution of lesions, but relapses and adverse effects are common. PMID- 15789046 TI - Oral lycopene--an efficacious treatment for oral leukoplakia? AB - DESIGN: This was a randomised controlled trial (RCT) of the treatment of oral leukoplakia with the carotenoid lycopene. INTERVENTION: A total of 58 patients received either 8 mg oral lycopene in two doses daily (n=20), 4 mg oral lycopene in two doses daily (n=18) or placebo capsules (n=18), for a 3-month period. Progress of patients was followed for a further 2 months. OUTCOME MEASURES: An objective clinical response, evaluated by bidimensional measurement of the lesion and colour photography, was classified as complete, partial, stable or progression. Histological status was categorised and ranked as normal (0), atypical hyperplasia (1), mild dysplasia (2), moderate dysplasia (3) or severe dysplasia (4). Histological response was then described by the change in rank, for example, from moderate dysplasia (3) to atypical hyperplasia (1) would indicate an improvement of 2 units. RESULTS: There was no significant difference in the clinical response of people who took 8 mg lycopene compared with those taking 4 mg lycopene. The clinical responses measured in both these groups were significantly greater, however, than those in the control group (P<0.01). The response, assessed histologically, after the 8-mg lycopene treatment was significantly better than that from 4 mg lycopene (P<0.05) and than the response seen in the control group (P<0.001). Patients taking 4 mg lycopene also responded significantly better than those in the control group (P<0.05). CONCLUSIONS: Oral lycopene appears, from this small RCT conducted over 5 months, to be effective in the treatment and management of oral leukoplakia. PMID- 15789047 TI - Bidi smokers at increased risk of oral cancer. AB - DATA SOURCES: Source articles were searched for using Medline, the Cochrane Library and within the references lists of identified articles. STUDY SELECTION: Articles were selected that included data enabling construction of 2 x 2 tables to estimate odds ratios (OR) and 95% confidence intervals (CI). DATA EXTRACTION AND SYNTHESIS: For each study, two-way contingency tables were constructed, based on exposure frequency distributions, for cases and controls. Unadjusted OR and 95% CI were recalculated based on the reported data using standard procedures. Separate contingency tables were made for bidi smoking, cigarette smoking and both types of smoking if the data were available in the same article. The overall OR combined across all studies, and its 95% CI, was calculated using a random effects model for bidi and cigarette smoking. Tests for publication bias and heterogeneity were conducted. Confounding factors, for example, betel quid chewing or alcohol use, were not included in the meta-regression model. RESULTS: An increased risk of oral cancer was found for bidi smokers compared with people who had never smoked (OR, 3.1; 95% CI, 2.0-5.0) whereas no significant pattern of risk was found for cigarette smokers (OR, 1.1; 95% CI, 0.7-1.8). There was substantial heterogeneity in the pooled OR estimate. CONCLUSIONS: The results clearly indicate that bidi smokers are at increased risk of oral cancer. It is important that this information be incorporated into smoking prevention and cessation efforts, particularly in the urban poor and rural mass in south Asian countries where bidi smoking is widespread. PMID- 15789048 TI - Multiple idiopathic cervical root resorption most frequently seen in younger females. AB - DATA SOURCES: Medline and references of included studies were used to source articles. STUDY SELECTION: Selected articles were case reports or reviews of patients who had multiple root resorption with no systemic or local factors that may have contributed to root resorption, the resorption originating at the cemento-enamel junction of teeth and involving more than three teeth in the dentition. DATA EXTRACTION AND SYNTHESIS: Clinical, histological and historical findings are summarised for all identified patients. RESULTS: Multiple idiopathic cervical root resorption (MICRR) was an incidental finding on routine clinical and radiographic examination. There appeared to be no correlation between this type of resorption and any medical or dental finding. Radiographically, MICRR was found to begin at the cemento-enamel junction and then either progress to involve the entire cervical region or, at some point, spontaneously arrest. Those cases that progressed to involve the entire cervical region required extraction. The number of teeth that demonstrated this condition ranged from five up to 24 per patient. More teeth became involved as the condition was followed in time. There was no detectable frequency of occurrence for any particular dental region or tooth among the involved teeth. Of a total of 18 patients, 13 were females whose ages ranged from 7 to 68 years of age. Ten of the 18 patients were Caucasian. CONCLUSIONS: Multiple idiopathic cervical root resorption was found most frequently associated with younger females. This condition appeared to be of unknown aetiology and uncertain natural history. PMID- 15789049 TI - Longer orthodontic treatment may result in greater external apical root resorption. AB - DATA SOURCES: Medline was the primary data source, with references of identified papers then being reviewed for additional studies. STUDY SELECTION: Studies were included if they were clinical trials, in English, of samples of greater than 10 subjects who had undergone fixed appliance orthodontic treatment and who had both pre- and postoperative X-rays available. A measurement of external apical root resorption (EARR) in maxillary incisors was essential as was a measure of displacement with the apex of the root as reference. DATA EXTRACTION AND SYNTHESIS: Variables were coded and articles graded by three independent investigators who subsequently negotiated final coding and assessed the methodological soundness of each study. A cumulative 'meta-analysis factor' was computed for each article. Articles were evaluated on study design; population sample; treatment assignment; documentation of statistics; the accuracy of root resorption measurement and apical displacement of incisor roots. A funnel plot analysis did not identify publication bias. RESULTS: Eight articles were included in the statistical analysis. Two did not contain data for mean apical displacement and therefore correlations between mean EARR and mean apical displacement could not be calculated for them. The mean meta-analysis factor was 39 (range, 12-78). Mean root resorption was 1.421+/-0.448 mm (n=8), and mean apical displacement was 2.382+/-0.756 mm. The weighted correlation coefficient between mean root resorption and apical displacement was 0.822 and that between mean root resorption and treatment duration was 0.852. CONCLUSIONS: This analysis suggests that treatment-related root resorption is correlated with the distance the apex moves and the length of time the treatment took. PMID- 15789050 TI - More women in Europe and Australia have dental agenesis than their counterparts in North America. AB - DATA SOURCES: Medline and Embase were searched, using the following keywords: "hypodontia", "oligodontia", "anodontia", "agenesis", and "prevalence or incidence". Reference lists from retrieved articles were also examined. STUDY SELECTION: For inclusion, an article had to satisfy the following criteria: presence of an English abstract; the sample was representative for the underlying general population; the diagnosis of dental agenesis was based on a radiographic examination; the report presented information on the ethnic background; and the report presented prevalence of agenesis except third molars. DATA EXTRACTION AND SYNTHESIS: Data from Caucasian populations in North America, Australia and Europe were included in a meta-analysis. For the prevalence in African-American, Chinese and Arab groups, only indications could be reported because of a limited number of studies. Multiple-regression analysis was applied to evaluate the influence of chronological age, sample size, continent and year of publication. Statistical significance was established at P<0.05. The prevalence of agenesis per tooth type, affected patients and number of missing teeth per patient was calculated as far as reported in the papers. For the comparison of the prevalence for males and females, the relative risk (RR) was calculated. RESULTS: The search identified 141 articles, of which 31 met the inclusion criteria. Agenesis differs by continent and gender: the prevalence for both sexes was higher in Europe (males 4.6%; females 6.3%) and Australia (males 5.5%; females 7.6%) than for North American Caucasians (males 3.2%; females 4.6%). In addition, the prevalence of dental agenesis in females was significantly higher than in males for all three continents, with an RR of 1.37 (95% CI, 1.28-1.45). The mandibular second premolar was the most affected tooth, followed by the maxillary lateral incisor and the maxillary second premolar. Unilateral occurrence of dental agenesis is more common than bilateral occurrence. Bilateral agenesis of maxillary lateral incisors is more common, however, than unilateral agenesis. In most patients, dental agenesis involved only one (48%) or two teeth (35%). CONCLUSIONS: The prevalence of dental agenesis in Europe and Australia is higher than in North America. The overall prevalence of agenesis in the maxilla is comparable with that in the mandible, but a marked difference was found between both jaws regarding tooth type. Absence of one or two permanent teeth is found in the majority of the subjects with dental agenesis. PMID- 15789051 TI - Grinding is effective in early orthodontic treatment of unilateral posterior crossbite. AB - DATA SOURCES: PubMed and the Cochrane Controlled Clinical Trials Register were searched. Reference lists from retrieved articles were also searched by hand for additional studies. STUDY SELECTION: For inclusion, an article had to satisfy the following criteria: cases in primary and early mixed dentition with unilateral posterior crossbite; be a randomised controlled trials (RCT) or prospective and retrospective studies with concurrent untreated as well as normal controls; clinical trials must have compared at least two treatment strategies without any untreated or normal control group involved; and be written in English, German, French or a Scandinavian language. DATA EXTRACTION AND SYNTHESIS: Data extraction was carried out independently and in duplicate. Quality of the studies was assessed and categorised as low, medium and high. Interexaminer conflicts were resolved by discussion to reach a consensus. RESULTS: The search identified 1001 articles, of which 12 met the inclusion criteria. Only two RCT of early treatment of crossbite have been performed, and these two studies support grinding as treatment in the primary dentition. As treatment strategies, quad-helix (QH), expansion plates and rapid maxillary expansion (RME) are effective in the early mixed dentition with high success rates. There is, however, no scientific evidence available that shows which of the treatment modalities, that is, grinding, QH, expansion plates or RME, is the most effective. CONCLUSIONS: Most of the studies have the serious drawback of lack of power because of small sample size, bias and confounding variables, lack of method error analysis and blinding in measurements, and deficient or lack of statistical methods. Thus, the studies were not of high enough quality level to draw any evidence-based conclusions. Better-controlled RCT with sufficient sample sizes are needed to obtain reliable scientific evidence identifying which treatment is the most effective for early correction of unilateral posterior crossbite. Future studies should also include assessments of long-term stability as well as analysis of costs and side-effects of the interventions. PMID- 15789055 TI - Disease association of two distinct interleukin-18 promoter polymorphisms in Caucasian rheumatoid arthritis patients. AB - Interleukin (IL)-18 is an important mediator of innate and adaptive immunity. We searched for an association of IL-18 promoter single-nucleotide polymorphisms (SNP) with rheumatoid arthritis (RA) in Caucasians. The entire study population was composed of two independent cohorts from Germany (n=200) and Scotland (n=410). Presence of IL-18 SNP at positions -607 and -137 was determined by allele-specific PCR in 327 RA patients and 283 healthy donors (HD). Diplotype distributions of both loci were in Hardy-Weinberg equilibrium (HWE) in the German and Scottish HD cohorts. In contrast, locus -607 was in HW disequilibrium in German, and locus -137 in Scottish RA patients. Diplotypic exact chi(2) tests suggested that -607CC was overrepresented in German, and -137CC in Scottish RA patients, but conservative chi(2) trend analyses could not prove any significant disease association of these single loci. SNP -607 and -137 were in strong linkage disequilibrium. The -607C(*)-137C haplotype was more prevalent in German RA (3.2 vs 1.2%) and in Scottish RA patients (4.1 vs 0.9%) than in the respective HD cohorts. These observations suggest that SNP of both positions contribute to the genetic background of RA pathogenesis. PMID- 15789056 TI - Risk of trachomatous scarring and trichiasis in Gambians varies with SNP haplotypes at the interferon-gamma and interleukin-10 loci. AB - Experimental evidence implicates interferon gamma (IFNgamma) in protection from and resolution of chlamydial infection. Conversely, interleukin 10 (IL10) is associated with susceptibility and persistence of infection and pathology. We studied genetic variation within the IL10 and IFNgamma loci in relation to the risk of developing severe complications of human ocular Chlamydia trachomatis infection. A total of 651 Gambian subjects with scarring trachoma, of whom 307 also had potentially blinding trichiasis and pair-matched controls with normal eyelids, were screened for associations between single-nucleotide polymorphisms (SNPs), SNP haplotypes and the risk of disease. MassEXTEND (Sequenom) and MALDI TOF mass spectrometry were used for detection and analysis of SNPs and the programs PHASE and SNPHAP used to infer haplotypes from population genetic data. Multivariate conditional logistic regression analysis identified IL10 and IFNgamma SNP haplotypes associated with increased risk of both trachomatous scarring and trichiasis. SNPs in putative IFNgamma and IL10 regulatory regions lay within the disease-associated haplotypes. The IFNgamma +874A allele, previously linked to lower IFNgamma production, lies in the IFNgamma risk haplotype and was more common among cases than controls, but not significantly so. The promoter IL10-1082G allele, previously associated with high IL10 expression, is in both susceptibility and resistance haplotypes. PMID- 15789057 TI - Renin-angiotensin system gene polymorphisms predict the progression to renal insufficiency among Asians with lupus nephritis. AB - The renin-angiotensin system (RAS) is a strong candidate as a mediator for the development and progression of lupus nephritis (LN). We performed an ethnically stratified analysis of 642 systemic lupus erythematosus (SLE) patients to determine whether various functional RAS gene polymorphisms are associated with SLE renal outcomes. Patients were genotyped for two angiotensin-converting enzyme (ACE) gene polymorphisms: Alu insertion/deletion (I/D) and 23 949 (CT)(2/3), and for two angiotensinogen (Atg) gene polymorphisms: M235T and C-532T. Multivariate analyses demonstrated associations between the ACE I/D, ACE (CT)(2/3) and Atg C 532T functional polymorphisms and LN among Asians. In stratified analyses among LN cases according to high vs low glomerular filtration rate (GFR), associations remained significant for the ACE D (odds ratio (OR) 5.9, P=0.001) and (CT)(2) (OR 6.2, P=0.001) alleles among Asian subjects with low GFR. Lastly, we found allelic dose-dependent associations between the ACE I/D (P=0.003), ACE (CT)(2/3) (P=0.005) and Atg M235T (P=0.04) polymorphisms, and GFR analyzed as a continuous variable among Asians. These findings suggest a significant role for ACE and Atg gene sequence variation and severity of LN among Asians with SLE. PMID- 15789058 TI - Immune response in silico (IRIS): immune-specific genes identified from a compendium of microarray expression data. AB - Immune cell-specific expression is one indication of the importance of a gene's role in the immune response. We have compiled a compendium of microarray expression data for virtually all human genes from six key immune cell types and their activated and differentiated states. Immune Response In Silico (IRIS) is a collection of genes that have been selected for specific expression in immune cells. The expression pattern of IRIS genes recapitulates the phylogeny of immune cells in terms of the lineages of their differentiation. Gene Ontology assignments for IRIS genes reveal significant involvement in inflammation and immunity. Genes encoding CD antigens, cytokines, integrins and many other gene families playing key roles in the immune response are highly represented. IRIS also includes proteins of unknown function and expressed sequence tags that may not represent genes. The predicted cellular localization of IRIS proteins is evenly distributed between cell surface and intracellular compartments, indicating that immune specificity is important at many points in the signaling pathways of the immune response. IRIS provides a resource for further investigation into the function of the immune system and immune diseases. PMID- 15789059 TI - Selective induction of tumor-associated antigens in murine pulmonary vasculature using double-targeted adenoviral vectors. AB - Targeted therapies directed to tumor-associated antigens are being investigated for the treatment of cancer. However, there are few suitable animal models for testing the ability to target these tumor markers. Therefore, we have exploited mice transgenic for the human coxsackie and adenovirus receptor (hCAR) to establish a new model for transient expression of human tumor-associated antigens in the pulmonary vasculature. Systemic administration of Ad in hCAR mice resulted in an increase in transgene expression in the lungs compared to wild-type mice, as determined using a luciferase reporter gene. To reduce transgene expression in the liver, the predominant organ of ectopic Ad localization and transgene expression following systemic administration, we utilized the endothelial specific flt-1 promoter, which resulted in a further increased lung-to-liver ratio of luciferase expression. Administration of an adenoviral vector encoding the tumor-associated antigen carcinoembryonic antigen (CEA) under transcriptional control of the flt-1 promoter resulted in selective expression of this antigen in the pulmonary vasculature of hCAR mice. Feasibility of targeting to expressed CEA was subsequently demonstrated using adenoviral vectors preincubated with a bifunctional adapter molecule recognizing this tumor-associated antigen, thus demonstrating utility of this transient transgenic animal model. PMID- 15789060 TI - GCV phosphates are transferred between HeLa cells despite lack of bystander cytotoxicity. AB - The role of gap junctional intercellular communication (GJIC) in bystander killing with herpes simplex virus thymidine kinase (HSV-TK) and ganciclovir (GCV) was evaluated in U251 cells expressing a dominant-negative connexin 43 cDNA (DN14), and in HeLa cells, reportedly devoid of connexin protein. These cell lines both exhibited 0% GJIC when assayed by Lucifer Yellow fluorescent dye microinjection. Bystander cytotoxicity was still apparent in 50:50 cocultures of DN14 and HSV-TK-expressing U251 cells, but not in 50:50 cocultures of HeLa cells. However, the sensitivity of HeLa HSV-TK-expressing cells to GCV decreased nearly 100-fold (IC90=109 microM) when cocultured with bystander cells compared to results in 100% cultures of HSV-TK-expressing cells (IC90=1.2 microM). A more sensitive flow cytometry technique to measure GJIC over 24 h revealed that the DN14 and HeLa cells exhibited detectable levels of communication (29 and 23%, respectively). Transfer of phosphorylated GCV to HeLa bystander cells occurred within 4 h after drug addition, and GCV triphosphate (GCVTP) accumulated to 213+/ 84 pmol/10(6) cells after 24 h. In addition, GCVTP levels were decreased in HSV TK-expressing cells in coculture (867+/-33 pmol/10(6) cells) compared to 100% cultures of HSV-TK-expressing cells (1773+/-188 pmol/10(6) cells). The half-life of GCVTP in the HSV-TK-expressing cells was approximately four times that measured in the bystander cells (12.3 and 3.1 h, respectively). These data suggest that the lack of bystander cytotoxicity in HeLa cocultures is due to low transfer of phosphorylated GCV and a rapid half-life of GCVTP in the bystander cells. Thus, GCV phosphate transfer to non-HSV-TK-expressing bystander cells may mediate either bystander cell killing or sparing of HSV-TK-positive cells, depending upon the cell specific drug metabolism. PMID- 15789061 TI - Combined CD4+ Th1 effect and lymphotactin transgene expression enhance CD8+ Tc1 tumor localization and therapy. AB - Type 1 T cells are the major components in antitumor immunity. The lack of efficient CD8(+) cytotoxic T (Tc) cell infiltration of tumors is a major obstacle to adoptive Tc-cell therapy. We have previously demonstrated that adenovirus (AdV)-mediated transgene lymphotactin (Lptn) expression by intratumoral AdVLptn injection and intravenous CD4(+) helper T (Th) cell transfer can enhance Tc-cell tumor infiltration and eradication of early stage tumors (5 mm in diameter). In this study, we generated ovalbumin (OVA)-specific Tc1 and Th1 cells in vitro by incubation of OVA-pulsed dendritic cells with naive T cells from T-cell receptor (TCR) transgenic OT I and OT II mice. We then investigated the potential synergy of Th1 help effect and Lptn transgene expression in Tc1-cell therapy of well established OVA-expressing EG7 solid tumors (7 mm in diameter). Our data showed that a combined adoptive T-cell therapy of Th1 (2.5 x 10(6) cells per mouse) and Tc1 (5 x 10(6) cells per mouse) resulted in regression of all eight (100%) transgene Lptn expressed EG7 tumors, which is significantly higher than four from eight (50%) in AdVLptn/Tc1 group and two from eight (25%) in Tc1/Th1 group (P < 0.05). The amount of transferred Tc1 cells detected in Lptn-expressed tumors with Th1 treatment is 0.72%, which is significantly higher than those of AdVLptn (0.22%), Th1 (0.41%) and the control AdVpLpA (0.09%) treatment groups (P < 0.05). Enhanced Tc1 tumor localization may be derived from the chemotactic effect of Lptn and the proliferative effect of Th1 and Lptn. This novel therapeutic strategy with enhancement of Tc1 tumor localization in the therapy of well established tumors may become a tool of considerable conceptual interest in the implementation of future clinical objectives. PMID- 15789062 TI - Efficient, cardiac-specific adenoviral gene transfer in rat heart by isolated retrograde perfusion in vivo. AB - While a number of virus-based delivery schemes have been developed for myocardial gene transfer, no technique has proven capable of modifying a majority of cardiac myocytes rapidly and homogeneously in the in vivo rat model, and most schemes result in significant infection of the liver and other organs. However, adenoviral delivery to the excised heart during retrograde perfusion can produce 67-92% efficient gene transfer. In this study, we adapt this isolation/perfusion scheme to the heart in vivo. We isolated the heart in vivo by simultaneously clamping all vessels to/from the heart. The heart was then continuously retrograde perfused through a catheter positioned in the aortic root. A second catheter in the right ventricle provided a path for efflux. After perfusing the heart for 7.5 min with calcium-free Tyrode solution followed by 90 s no-flow viral exposure (AdV.cmv.LacZ; 10(12) viral particles/ml), gene transfer efficiency was 60% compared to 5% by a conventional cross-clamp technique. Infection of peripheral organs was dramatically reduced. Given the prevalence of the rat in so many models of heart disease, this enhancement of infection represents an advancement in viral-based delivery of exogenous genes to heart for the study of gene therapy in vivo. PMID- 15789063 TI - Nonviral ocular gene transfer. AB - In this study, we explored the use of electroporation or media that promote lipoplex formation for nonviral gene transfer in the eye. There was no detectable staining for LacZ after subretinal, intravitreous, or periocular injection of a plasmid containing a CMV promoter expression cassette for LacZ, but when plasmid injection in each of the three sites was combined with electroporation, there was efficient transduction. Specific staining for LacZ was seen in retinal pigmented epithelial (RPE) cells after subretinal injection of a plasmid containing a vitelliform macular dystrophy 2 (VMD2) promoter expression cassette, demonstrating that this approach can be used to evaluate purported tissue specific promoters in vivo. Electroporation with 10 V/mm resulted in strong LacZ staining, but was damaging to photoreceptors; substantial transduction with no evidence of retinal damage was seen using 3.4 V/mm. Staining for LacZ was also seen after subretinal or periocular, but not intravitreous, injection of plasmid DNA in medium containing 40% Lipofectamine2000 (Lf). Injection of 40% Lf into the subretinal space caused damage to photoreceptors, but subretinal injection of plasmid DNA in medium containing 10% NeuroPorter resulted in transduction of RPE cells with no adverse effects on retinal morphology or function as assessed by electroretinograms (ERGs). After either electroporation or lipofection, LacZ staining was detectable for at least 14 days, and could be reinduced by a second procedure. These data suggest that electroporation or lipofection can be used as experimental tools for ocular gene transfer to evaluate tissue-specific promoter fragments or to evaluate the effects of transgene expression in the retina. Also, with additional optimization, nonviral gene transfer may prove to be a valuable approach for the treatment of retinal and choroidal diseases. PMID- 15789064 TI - Full kringles of plasminogen (aa 1-566) mediate complete regression of human MDA MB-231 breast tumor xenografted in nude mice. AB - Since kringle (K)5, not present in the angiostatin molecule, was shown to be a key functional domain possessing potent antiangiogenic activity, we have evaluated a new plasminogen-derived fragment, consisting of the N-terminal part of human plasminogen, that included the complete secondary structure of K1-5 (aa 1-566). In contrast to other fragments described to date, K1-5 includes cysteine residues at positions 543, 555 and 560 allowing the formation of the three disulfide bonds lying within K5. Vascular endothelial cell proliferation and migration assays revealed that a replication-defective adenovirus (AdK1-5(1 566)), expressing K1-5 (aa 1-566), was dose dependently more potent that AdK1-3(1 354), an adenovirus that expresses only the first three kringles. In contrast to AdK1-3(1-354), a single intratumoral injection of AdK1-5(1-566) into MDA-MB-231 breast human carcinoma tumors was followed by a total regression of 40% of the tumor and by significant arrest of tumor growth (90%), which was correlated with a drastic decrease of functional neovascularization into the tumors. Furthermore, systemic delivery of AdK1-5(1-566) in mice inhibited the lung invasion of melanoma B16-F10 cells by 87%. Our findings provide evidence that the full kringles of plasminogen (aa 1-566) may be much more potent than K1-3 (aa 1-354), for the suppression of angiogenesis, tumor growth and metastatic dissemination. PMID- 15789065 TI - Untreated essential thrombocythemia evolving to biphenotypic leukemia, Philadelphia chromosome positive with monosomy 7: response to imatinib and reduced-intensity allogeneic stem cell transplant. PMID- 15789066 TI - Genetic heterogeneity of BCR/ABL+ adult B-cell precursor acute lymphoblastic leukemia: impact on the clinical, biological and immunophenotypical disease characteristics. AB - Philadelphia-positive (Ph(+)) B-cell precursor acute lymphoblastic leukemia (BCP ALL) is a genetically heterogeneous disease with a very poor prognosis. In this study, we analyzed the frequency of supernumerary Ph, trisomy 8, monosomy 7, and del(9p21) by FISH and its relationship with the characteristics of the disease, in 46 BCR/ABL(+) adult BCP-ALL patients. The frequency of supernumerary Ph, trisomy 8, monosomy 7 and del(9p21) was 30%, 20%, 15%, and 24%, respectively. Although all patients displayed a BII/common phenotype, supernumerary Ph and trisomy 8 were associated with higher expression of CD19 and CD22 and of CD19, CD34, CD45, and HLA-DR, respectively; in turn, cases with monosomy 7 showed lower CD19, CD22, CD34, and cCD79a and del(9p21)(+) blasts were CD13(-) and CD33(-). Overall, similar clinical and hematological features were observed at presentation, independently of the underlying genetic abnormalities. However, relapse-free survival (RFS) was significantly shorter in cases with supernumerary Ph, trisomy 8, and del(9p21), the latter being the most powerful independent prognostic factor for RFS. PMID- 15789067 TI - Broad molecular screening of an unclassifiable myeloproliferative disorder reveals an unexpected ETV6/ABL1 fusion transcript. PMID- 15789068 TI - Flow cytometry evaluation of erythroid and myeloid dysplasia in patients with myelodysplastic syndrome. AB - The purpose of this study was to develop a flow cytometric approach to the evaluation of marrow dysplasia in myelodysplastic syndromes (MDS). We first studied a cohort of 103 MDS patients as well as 46 pathological and healthy controls. Flow cytometry data were expressed as percentage of positive cells. Analysis of erythroid cells showed higher proportions of immature cells (P < 0.001) and decreased levels of CD71 expression on nucleated red cells (P = 0.02) in MDS. Analysis of myeloid cells showed lower proportions of CD10+ and higher proportions of CD56+ granulocytes (P < 0.001), and increased ratios of immature to mature cells (P = 0.007). Since no single immunophenotype could accurately differentiate MDS from other conditions, we used discriminant analysis for generating erythroid and myeloid classification functions using combinations of immunophenotypic parameters. These functions were prospectively validated in a testing cohort of 69 MDS patients and 46 pathological controls. A diagnosis of MDS was obtained in 60/69 cases (87%). No false-positive results were noticed among controls. Significant correlations between values of these functions and both degree of morphological dysplasia and the International Prognostic Scoring System were found. These findings indicate that flow cytometry evaluation of marrow dysplasia is feasible and may be useful in the work-up of individual MDS patients. PMID- 15789069 TI - High concordance from independent studies by the Children's Cancer Group (CCG) and Pediatric Oncology Group (POG) associating favorable prognosis with combined trisomies 4, 10, and 17 in children with NCI Standard-Risk B-precursor Acute Lymphoblastic Leukemia: a Children's Oncology Group (COG) initiative. AB - Chromosome aberrations have a major role in pediatric acute lymphoblastic leukemia (ALL) risk assignment. The Children's Cancer Group (CCG) and the Pediatric Oncology Group (POG) independently assessed the significance of trisomy for chromosomes 4, 10, and 17 in National Cancer Institute (NCI) Standard- and High-Risk ALL. Data from 1582 (CCG) and 3902 (POG) patients were analyzed. Eight year event-free survivals (EFS) of 91% (CCG) and 89% (POG) (P < 0.001) were achieved in patients assigned to NCI Standard Risk whose leukemic cells had simultaneous trisomies 4, 10, and 17. Both groups showed the degree of favorable prognostic importance increased with the actual number of favorable trisomies. POG analyses also demonstrated hyperdiploidy (> or =53 chromosomes) was less of an independently significant prognostic factor in the absence of these key trisomies. This finding supported conclusions from previous CCG and POG studies that specific trisomies are more important than chromosome number in predicting outcome in pediatric B-precursor ALL. In NCI Higher Risk patients, the number of favorable trisomies was not prognostically significant, but showed the same trend. Moreover, specific trisomies 4, 10, and 17 remain associated with favorable prognosis in Standard-Risk B-precursor ALL, even in the context of very different treatment approaches between the groups. PMID- 15789071 TI - Lifestyle dentistry. PMID- 15789072 TI - Application packs. PMID- 15789073 TI - Evidence based dentistry. PMID- 15789074 TI - Managing change. PMID- 15789075 TI - A mouthful of trouble. PMID- 15789086 TI - Antibiotic prophylaxis and third molar surgery. AB - The issue of prophylactic antibiotic therapy in third molar surgery is highly controversial. The current evidence questions the benefits of routine prophylactic antibiotic therapy which does not appear to overcome the risk of undesirable outcomes after third molar removal. In our opinion there is no justification for routine antibiotic prophylaxis for third molar surgery. PMID- 15789087 TI - Palatal and nasal necrosis resulting from cocaine misuse. AB - The potential social and economic harm caused by cocaine addiction is well known. The wider health risks and potential nasal complications of septal perforation and nasal collapse are also recognised. We report a case of oro-nasal fistula secondary to cocaine misuse and discuss the aetiology, management and potential methods of repair. PMID- 15789088 TI - Recurrent sinus of the cheek due to a retained foreign body: report of an unusual case. AB - Pathological sinuses in the maxillofacial region are frequently encountered in clinical practice. These sinuses may be a sequelae of periapical and periodontal pathologies or infections like osteomyelitis, actinomycosis etc. Classical clinical symptoms and radiographic features accompany all these infections. Rarely, sinuses in the oro-facial region can be sequelae of retained occult foreign bodies like wood in the soft tissues. We report a case of recurrent sinus of the cheek caused by an occult wooden splinter and discuss its diagnosis and clinical management. PMID- 15789089 TI - An undiagnosed case of malignancy: case report. AB - The following report describes the treatment carried out by a UK high street dental technician on an elderly patient with a large malignancy invading the oral cavity through the hard palate from the nose. The lesion remained undiagnosed during treatment for three months. The consequences of this error, and the future role of the clinical dental technician are discussed. PMID- 15789097 TI - Clinical trials. PMID- 15789098 TI - Steroid cover. PMID- 15789099 TI - Rubber dams. PMID- 15789100 TI - Chronic pain. PMID- 15789101 TI - Indices. PMID- 15789102 TI - An investigation of the efficacy of super-oxidised (Optident/Sterilox) water for the disinfection of dental unit water lines. AB - AIMS: To determine the efficacy of super-oxidised water (Optident/Sterilox) in the decontamination of dental unit water lines. METHODS: Dental units (10) were first purged with concentrated super-oxidised water. After purging, a 5% (v/v) super-oxidised water was used as a maintenance dose. Samples for microbiology were taken after 0,1,2,3,4,5,6,7 d, and each week for a further 13 weeks. RESULTS: After purging, 5% (v/v) super-oxidised water was successful in reducing the microbial counts to zero, although in three of the units some bacteria were intermittently isolated in the first week of treatment. CONCLUSIONS: Super oxidised water was successful in the removal of bacteria from dental unit water supplies. Complete removal required the treatment with a purge phase of concentrated disinfectant and a maintenance phase of at least two weeks. PMID- 15789103 TI - A 3-year longitudinal, controlled clinical study of a gallium-based restorative material. AB - AIM: The aim of this three-year longitudinal controlled study was to compare the clinical performance of Galloy versus a high copper, mercury based Dispersalloy filling material. METHODS: Moderate to large class I and class II cavities or replacement restorations were selected and 25 Galloy restorations and 25 Dispersalloy controls were placed in 14 adult patients by a single operator. Restorations were photographed and a silicone impression recorded at baseline, 6 months, 1 year, 2 years and 3 years. RESULTS: At 3 years all 22 Dispersalloy restorations but only 4 Galloy restorations were still in situ. Three Dispersalloy restorations were lost to follow-up. Tooth fracture was observed in 15 (60%) of the Galloy restorations by the end of the 3 years, compared to one (4%) Dispersalloy restoration, which failed due to tooth fracture. A further six Galloy restorations had to be removed due to complaints of persistent pain. Four teeth restored with Galloy required endodontic treatment but none of the Dispersalloy restored teeth required endodontics. Of the four Galloy restorations remaining in situ, three were relatively small restorations and the fourth a moderate sized restoration required a marginal repair. CONCLUSION: The clinical performance of Galloy restorations was so grossly inferior to the Dispersalloy controls that Galloy cannot be recommended for clinical use in moderate to large or multi-surface cavities. PMID- 15789104 TI - Oral lichenoid lesions related to dental restorative materials. AB - OBJECTIVES: To determine the effectiveness of replacing restorations considered to be the cause of an oral lichenoid lesion (oral lichenoid reaction)(OLL). DESIGN: Clinical intervention and nine-month follow up. SETTING: The study was carried out in the University Dental Hospital of Manchester, 1998-2002. SUBJECTS AND METHODS: A total of 51 patients, mean age 53 (SD 13) years, who had oral lesions or symptoms suspected to be related to their dental restorations were investigated. Baseline patch tests for a series of dental materials, biopsies and photographs were undertaken. Thirty-nine out of 51 (76%) of patients had their restorations replaced. RESULTS: The clinical manifestations of OLL were variable; the majority of OLL were found to be in the molar and retro molar area of the buccal mucosa and the tongue. Twenty-seven (53%) patients had positive patch test reactions to at least one material, 24 of them for one or more mercury compound. After a mean follow up period of nine months, lesions adjacent to replaced restorations completely healed in 16 (42%) patients (10 positive and 6 negative patch tests). Improvement in signs and symptoms were found in 18 (47%) patients (11 positive and 7 negative patch tests). CONCLUSION: OLLs may be elicited by some dental restorations. Replacing restorations adjacent to these lesions is associated with healing in the majority of cases particularly when lesions are in close contact with restorations. A patch test seems to be of limited benefit as a predictor of such reactions. PMID- 15789105 TI - Delivery of postgraduate dental education to meet the needs of general dental practice --a case study. AB - In order for dental practices to gain the maximum benefit from an educational opportunity such as a course or workshop, it is important that the training received meets the needs of these practices and is delivered appropriately. This paper provides an account of an educational initiative undertaken within the Eastern deanery in 2003 and 2004, 'Essex Promoting Infection Control' (EPIC), which set out to establish training needs in infection control within general dental practices across Essex and then to design and deliver learning events to meet these needs. PMID- 15789118 TI - Posts and the root-filled tooth. AB - It is clear that teeth often require significant reconstruction following root canal treatment. It is also evident that there is an enormous variety of ways that can be used to achieve this. Although there is some debate about the relative contributions of the root filling and coronal restoration to endodontic success, it is agreed that the best results are gained with a good root filling and a well-sealed coronal restoration. Posts, pins and/or bonding are all possible solutions to retain the coronal restoration but it is often difficult to decide, on the basis of evidence rather than fashion or anecdote, which is preferable. PMID- 15789121 TI - How molecular profiling could revolutionize drug discovery. AB - Information from genomic, proteomic and metabolomic measurements has already benefited target discovery and validation, assessment of efficacy and toxicity of compounds, identification of disease subgroups and the prediction of responses of individual patients. Greater benefits can be expected from the application of these technologies on a significantly larger scale; by simultaneously collecting diverse measurements from the same subjects or cell cultures; by exploiting the steadily improving quantitative accuracy of the technologies; and by interpreting the emerging data in the context of underlying biological models of increasing sophistication. The benefits of applying molecular profiling to drug discovery and development will include much lower failure rates at all stages of the drug development pipeline, faster progression from discovery through to clinical trials and more successful therapies for patient subgroups. Upheavals in existing organizational structures in the current 'conveyor belt' models of drug discovery might be required to take full advantage of these methods. PMID- 15789122 TI - Cellular processing of platinum anticancer drugs. AB - Cisplatin, carboplatin and oxaliplatin are platinum-based drugs that are widely used in cancer chemotherapy. Platinum-DNA adducts, which are formed following uptake of the drug into the nucleus of cells, activate several cellular processes that mediate the cytotoxicity of these platinum drugs. This review focuses on recently discovered cellular pathways that are activated in response to cisplatin, including those involved in regulating drug uptake, the signalling of DNA damage, cell-cycle checkpoints and arrest, DNA repair and cell death. Such knowledge of the cellular processing of cisplatin adducts with DNA provides valuable clues for the rational design of more efficient platinum-based drugs as well as the development of new therapeutic strategies. PMID- 15789123 TI - Time-course of morphologic changes and peptide YY adaptation in ileal mucosa after loop ileostomy in humans. AB - PURPOSE: The secretion mechanism of peptide YY involves systemic factors, such as humoral and neural stimuli, and local factors, such as intestinal peristalsis and intraluminal nutrients. This study was designed to survey the impact of local stimuli on the secretion of peptide YY under circumstances in which systemic stimuli are identical. METHODS: Ileostomies were repaired within three months in a short-term group (14 patients) and after six months in a long-term group (14 patients). Mucosal peptide YY concentrations and cytomorphologic change, such as villus height, crypt depth, mucosal thickness, and villus index, were compared between proximal functioning ileum and a distal nonfunctioning ileal loop during ileostomy repair. In a control group of patients undergoing right hemicolectomy (21 cases), the normal distribution of peptide YY was measured in the mucosa throughout the distal ileum. RESULTS: The peak of peptide YY concentration in the terminal ileum was 307.4 +/- 21 pmol/g, 25 cm from the ileocecal valve, with lower levels both proximally and distally. The mucosa of the functioning ileum in the short-term group showed hypertrophy, but had returned to preoperative levels in the long-term group. The nonfunctioning mucosa in both groups underwent atrophic changes. The mucosal peptide YY content in functioning ileum in the short-term group was higher than that of distal nonfunctioning mucosa (363.9 +/- 25.5 pmol/g vs. 284.1 +/- 13 pmol/g, P < 0.05), suggesting adaptive upregulation. The increments of mucosal peptide YY content in this short-term group compared with the control group were 45.6 and 4.7 percent in the proximal and distal segments, respectively. In the long-term group, proximal and distal mucosal peptide YY were similar (256.6 +/- 31.9 pmol/g vs. 254.9 +/- 27.1 pmol/g, P > 0.05), and there was no increment in either (1.3 vs. 4.4 percent, P > 0.05). CONCLUSIONS: The peak concentrations of PYY in the ileal mucosa are found 20 to 25 cm proximal to the ileocecal valve. In the short-term response of ileostomy, local stimulatory factors play a major role in the adaptation of mucosal PYY. In the defunctioned bowel without luminal stimulation, systemic stimulation was important for maintenance of mucosal PYY. PMID- 15789124 TI - Reduction of a large incarcerated rectal prolapse by use of an elastic compression wrap. AB - Reduction of a large rectal prolapse may be difficult because of significant edema that collects in the rectal tissues. If reduction is unsuccessful, an emergent laparotomy and internal reduction is required. A wide elastic wrap applied around the prolapsed rectum provides progressive compression, which reduces the amount of edema, allowing subsequent manual reduction. This novel technique is simple, safe, inexpensive, and can easily be performed in the emergency department setting. Manual reduction, by this or other described methods, should be attempted before emergent laparotomy for incarcerated rectal prolapse is performed. PMID- 15789125 TI - Prognosis after anastomotic leakage in colorectal surgery. AB - INTRODUCTION: Anastomotic leakage is a major complication of colorectal surgery causing a significant increase in 30-day mortality. The long-term prognosis of anastomotic leakage is poorly documented. This study was designed to assess whether anastomotic leakage affects five-year survival and local recurrence. METHODS: A total of 5,173 patients were recruited to the Wessex Colorectal Cancer Audit during the period September 1991 to August 1995 (prospective data, 5-year follow-up). The effect of anastomotic leakage on five-year survival and local recurrence was analyzed using Kaplan-Meier curves and the log-rank test. RESULTS: A total of 1,834 patients underwent a curative resection with an anastomosis (anastomotic leak = 71; 3.9 percent): 30-day mortality: 18.3 percent in the leak group, and 3.5 percent in the nonleak group (P < 0.001); local recurrence: 19 percent in the leak group, and 9.8 percent in the nonleak group (P = 0.018). A total of 1,201 patients underwent colonic anastomosis (anastomotic leak = 31; 2.6 percent). There was no significant difference in local recurrence or five-year survival between the leak and nonleak groups. A total of 633 patients underwent rectal anastomosis (anastomotic leakage = 40; 6.3 percent): 30-day mortality: 10 percent in the leak group, and 2 percent in the nonleak group (P = 0.014); cumulative five-year estimate of local recurrence: 25.1 (95 percent confidence interval, 9.6-40.5) percent in the leak group, and 10.4 (95 percent confidence interval, 7.7-13) percent in the nonleak group (P = 0.007). Cumulative five-year estimate of overall survival: 52.8 (95 percent confidence interval, 36.1-69.4) percent in the leak group, and 63.9 (95 percent confidence interval, 59.9-67.9) percent in the nonleak group (P = 0.19). CONCLUSIONS: After rectal anastomosis, an anastomotic leak is associated with a significant increase in local recurrence. PMID- 15789126 TI - Isolated blood-cerebrospinal fluid barrier dysfunction: prevalence and associated diseases. AB - OBJECTIVE: An isolated dysfunction of the blood-CSF barrier is characterised by an abnormal elevation of the albumin CSF/serum concentration ratio (Q(alb)) without any other pathological CSF findings. Although common in routine CSF analysis, the clinical significance of an isolated barrier dysfunction frequently remains unclear. We examined neurological disorders associated with an isolated elevation of Q(alb) to identify possible determinants of blood-CSF barrier dysfunction. METHODS: 367 patients (124 women, 243 men, median age 60. 0 years) out of 3,873 patients receiving diagnostic lumbar puncture at the University Hospital of Ulm (Germany) showed an isolated dysfunction of the blood-CSF barrier. Clinical data as well as MRI findings of these patients were analysed. RESULTS: Isolated barrier dysfunction occurred most frequently (> 30%) in Guillain-Barre syndrome (GBS), chronic inflammatory demyelinating polyneuropathy (CIDP), normal pressure hydrocephalus (NPH), lumbar spinal stenosis, and polyneuropathy (PNP). In patients who showed no other evidence of neurological disease, isolated barrier dysfunction was found in 14. 9% of cases. The extent of barrier dysfunction was most prominent in brain tumours, GBS, and CIDP. There was a significant correlation of Q(alb) with both weight and body mass index (BMI). CONCLUSIONS: Although isolated barrier dysfunction may be found in a variety of neurological diseases, it is especially frequent in GBS, CIDP, NPH, spinal canal stenosis, and PNP. In these patients, disease-related mechanisms contributing to barrier dysfunction are likely. Moreover, barrier function seems to be influenced by disease-independent determinants like weight and BMI. PMID- 15789127 TI - Development and evaluation of the Parkinson Psychosis Questionnaire A screening instrument for the early diagnosis of drug-induced psychosis in Parkinson's disease. AB - OBJECTIVE: Drug-induced psychosis is a frequent side-effect in the treatment of advanced Parkinson's disease (PD). We sought to develop and evaluate a brief instrument for early recognition of drug-induced psychosis in PD. METHODS: We developed the "Parkinson Psychosis Questionnaire" (PPQ), which consists of screening questions for typical early signs and psychotic symptoms in PD and which quantifies the frequency and severity of four clinical categories-sleep disturbances, hallucinations/illusions, delusions and orientation. We performed an internal validation of the PPQ in 50 unselected patients with parkinsonism. The Brief Psychiatric Rating Scale (BPRS) and the "Structurized Clinical Interview" (SCID) for DSM IV were applied to the same patients as external references. RESULTS: Of 50 subjects, 49 suffered from idiopathic PD and one from probable MSA-P. Hoehn and Yahr stages in "on" ranged from 1.5 to 4. Sensitivity of the PPQ test for drug-induced psychosis according to SCID was 100 % (95 % CI: 73.5%, 100%); while specificity was 92.1 % (95% CI: 78.6%, 98.3 %). The PPQ severity score was highly correlated with BPRS. We derived a linear prediction formula, which transformed PPQ into BPRS scores. CONCLUSION: The PPQ appears to be a suitable, and easily administered instrument for early diagnosis of drug induced psychosis in routine PD care. Whether the PPQ could also be a valuable tool for monitoring follow-up studies and therapeutic intervention trials remains to be tested. PMID- 15789128 TI - Neurosyphilis and paraneoplastic limbic encephalitis: important differential diagnoses. PMID- 15789129 TI - Spastic paraparesis as a manifestation of metabolic vitamin B12 deficiency: a case report. PMID- 15789130 TI - Clinical trials are too often founded on poor quality pre-clinical research. PMID- 15789131 TI - Medical treatment for nocardial brain abscesses case report. PMID- 15789132 TI - A possible spatial and temporal cluster of multiple sclerosis in the town of Linguaglossa, Sicily. AB - We carried out an epidemiological survey to determine prevalence and incidence of multiple sclerosis in the little town of Linguaglossa in the Province of Catania. We calculated prevalence rate as point prevalence at 1 January 2001 and incidence during 1991-2000. We studied the frequency of multiple sclerosis in the community of Linguaglossa in a population of 5,422 inhabitants in the 2001 census. The primary sources for the case ascertainment were the general practitioners of Linguaglossa, the local Italian Multiple Sclerosis Association and the neurological departments, Multiple Sclerosis Centers and private neurologists of the province of Catania. We considered as prevalent and incident cases all patients who satisfied the Poser's diagnostic criteria. We detected 11 patients with multiple sclerosis who had had the onset of disease on prevalent day (P.D.). The onset-adjusted prevalence rate was 203/100,000 (95% CI 107-352). Prevalence was higher in women (247/100,000) than in men (154/100,000). From 1991 to 2000, 10 subjects with MS had clinical onset of disease. The mean annual incidence risk was 18.2/100,000 (C. I. 95 % 5.9-42.5). Conversely in the same population prevalence on 1 January 1991 was 37/100,000 while the onset adjusted annual incidence risk during the previous decade (1981-1991) was 3.6/100,000. Prevalence and incidence rates of MS during the last decade in the little town of Linguaglossa are higher than those found in the same area during the previous ten years and also than those reported in other Sicilian and Italian surveys suggesting a possible cluster of MS. PMID- 15789133 TI - Intrathecal baclofen for spasticity in primary lateral sclerosis. PMID- 15789134 TI - A quantitative method for the assessment of intraepidermal nerve fibers in small fiber neuropathy. AB - OBJECTIVES: The purpose of this paper is to present an easy-to-use and reproducible morphometrical method of determining the density of intraepidermal nerve fibers (IENF) per epidermal area with the corresponding reference range of the IENF-counts. METHODS: Thirty patients and 22 controls were included in this study. The patients were divided into three groups: small-fiber (SFN), diabetic and demyelinating neuropathy. All subjects underwent punch skin biopsy. Specimens were fixed routinely in formalin and thereafter embedded in paraffin. Nerve fibers were revealed using immunoperoxidase staining with panaxonal antibody PGP 9.5. Using light microscopy, immunopositive nerves were counted morphometrically per epidermal area (NPEA) and, for comparison, per epidermal length (NPEL). RESULTS: Both the NPEA and NPEL estimates of SFN and diabetic neuropathy group differed significantly from those of control specimen (p < 0.001 and p < 0.001, Mann-Whitney test). Our method of counting, NPEA, shows a good correlation to NPEL (r = 0.945). CONCLUSIONS: IENF-counting by a new morphometric modification is reproducible and diagnostically sensitive and can easily be adopted in any laboratory familiar with the basic immunohistochemical methodology. The method is less dependent on costly technical support systems and seems to be less time consuming when compared with conventional methods for IENF-counting. PMID- 15789136 TI - A new LMNA mutation causing limb girdle muscular dystrophy 1B. PMID- 15789135 TI - SOD1 mutations in amyotrophic lateral sclerosis. Results from a multicenter Italian study. AB - Amyotrophic Lateral Sclerosis (ALS), the most common form among motoneuron diseases, is characterized by a progressive neurodegenerative process involving motor neurons in the motor cortex, brain stem and spinal cord. Sporadic (SALS) accounts for the majority of patients but in about 10% of ALS cases the disease is inherited (FALS), usually as an autosomal dominant trait. In the present study we show the results of a referred based multicenter study on the distribution of SOD1 gene mutations in the largest cohort of Italian ALS patients described so far. Two hundred and sixty-four patients (39 FALS and 225 SALS) of Italian origin were studied. In 7 out of 39 FALS patients we found the following SOD1 gene mutations: i) a new G12R missense mutation in exon 1, found in a patient with a slowly progressive disease course; ii) the G41S mutation, in four unrelated patients with rapidly progressive course complicated with cognitive decline in two of them; iii) the L114F mutation, in a patient with a slowly progressive phenotype; iv) the D90A mutation, in a heterozygous patient with atypical phenotype. In addition, in one SALS patient a previously reported synonymous variant S59S was identified. In 17 (3 FALS and 14 SALS) out of 264 patients (6.4 %) the polymorphism A-->C at position 34 of intron 3 (IVS3: + 34 A-->C) was found, and in one FALS patient a novel variant IVS3 + 62 T-->C was identified. The frequency of SOD1 gene mutations (17.9 %) in FALS cases was comparable with that found in other surveys with a similar sample size of ALS cases. No SOD1 gene mutations have been identified in SALS cases. Within FALS cases, The most frequent mutation was the G41S identified in four FALS. PMID- 15789137 TI - Three-dimensional in vivo displacements of the shoulder complex from biplanar radiography. AB - The goal of this study was to adapt roentgen photogrammetry to in vivo studies of shoulder skeletal motion during arm elevation in the scapular plane. Numerous in vitro and in vivo studies have been published describing shoulder bone movements. They involve plain radiographic measurements and utilize a three-dimensional (3D) approach. Measurements are either direct using pins implanted in bones, or indirect recording points on medical images. Roentgen photogrammetry locates points in space from two projections obtained from two different radiographic incidences. The technique has been applied in vivo by implanting metallic balls in bones. However, to be used as a standard clinical procedure, the technique must be adapted to be less invasive. In vivo photogrammetric reconstruction of known points in 3D space requires that the subject is strictly motionless between the successive radiographic exposures or that the exposures are obtained simultaneously. Methods used in this study were developed to allow subsequent exposures to be used for analysis. Numerical tools have been developed to align the two projections of a point in 3D space which have moved slightly between two successive exposures. The standard photogrammetric technique is completed by geometric modeling of the shoulder complex and humerus, and by the control of their mutual proximity at the level of joints. Bones are modeled as a set of simple volumes linked together using geometric shapes described by shape parameters. The coincidence between real bone contours and radiographic projections of the modeled bone gives the values of the shape parameters and the accurate location in space. Results focus on two different topics: errors related to the use of roentgen photogrammetry with successive exposures, and results obtained by applying roentgen photogrammetry to the in vivo shoulder complex. Results describing shoulder bone and joint displacements are presented for comparison with previously published results. The technique of roentgen photogrammetry can successfully be applied to patients. The radiographic protocol is simple, and data can be obtained easily and quickly from the digitized films. The data obtained from asymptomatic shoulders compared favorably with published values. Future research will focus on comparisons between kinematics of the symptomatic and asymptomatic contralateral limbs in volunteers. PMID- 15789138 TI - Anatomical variations of the flexor hallucis longus muscle and the consequences for tendon transfer. A cadaver study. AB - The aim of this study was to evaluate the occurrence of anatomical variations of the musculotendinous junction of the flexor hallucis longus muscle. Eighty cadaver specimens preserved according to Thiel's method were assessed. Following careful dissection, the distance between the musculotendinous junction and the bone-cartilage border of the distal tibia was determined. Three typical anatomical variations were found: (1) a long lateral and shorter medial muscle belly; (2) equal-length medial and lateral muscle bellies; (3) a long medial and shorter lateral muscle belly. As a special variation in two cases only one lateral muscle belly was found. When planning a flexor hallucis longus transfer, preoperative magnetic resonance tomography (MRT) should be carried out in order to guarantee sufficient coverage of an existing soft tissue defect. These anatomical characteristics have also to be considered when interpreting ultrasonic or MRT findings in this region. PMID- 15789139 TI - Effect of adding dietary L-lysine, L-threonine and L-methionine to a low gluten diet on urea synthesis in rats. AB - We have shown that urinary urea excretion increased in rats fed a low quality protein. The purpose of present study was to determine whether an addition of dietary limiting amino acids affected urea synthesis in rats fed a low gluten diet. Experiments were done on three groups of rats given diets containing 10% gluten, 10% gluten +0.5% L-lysine or 10% gluten+0.5% L-lysine, 0.2% L-threonine and 0.2% L-methionine for 10 d. The urinary excretion of urea, and the liver concentrations of serine and ornithine decreased with the addition of dietary L lysine, L-threonine and L-methionine. The fractional and absolute rates of protein synthesis in tissues increased with the treatment of limiting amino acids. The activities of hepatic urea-cycle enzymes was not related to the urea excretion. These results suggest that the addition of limiting amino acids for the low gluten diet controls the protein synthesis in tissues and hepatic ornithine and decline urea synthesis. PMID- 15789141 TI - Protein metabolism and strength performance after bovine colostrum supplementation. AB - This study was designed to determine the responses of muscle protein, serum amino acids, and strength performance to bovine colostrum supplementation in physically active men. The rest (R) group (n = 6) and the exercise (E) group (n = 6) carried out twice a 2-week experiment randomly assigned in a double-blind fashion with either placebo (PLA; consuming daily 20 g maltodextrin) or bovine colostrum (COL; consuming daily 20 g colostrum supplement) treatment with one month between. On the test day after the treatment period the measurements were carried out in fasting conditions and E carried out a strength training session (STS). The methods involved the infusion of ring-(2)H(5)-phenylalanine, femoral arterial and venous blood sampling, and biopsies from the vastus lateralis muscle. Serum concentration of essential amino acids during recovery was greater (p < 0.05) in the COL groups compared with the PLA groups. Both muscle protein synthesis and breakdown increased (p < 0.05) with COL. There were no differences in phenylalanine net balance or strength performance between the PLA and COL groups. It was concluded that a 2-week supplementation with bovine colostrum in physically active men increases serum concentration of essential amino acids but has no effect either on strength performance or protein net balance in fasting conditions during recovery after STS. PMID- 15789140 TI - Effect of hyperprolinemia on acetylcholinesterase and butyrylcholinesterase activities in rat. AB - We observed here that acute proline (Pro) administration provoked a decrease (32%) of acetylcholinesterase (AChE) activity in cerebral cortex and an increase (22%) of butyrylcholinesterase (BuChE) activity in the serum of 29-day-old rats. In contrast, chronic administration of Pro did not alter AChE or BuChE activities. Furthermore, pretreatment of rats with vitamins E and C combined or alone, N(omega)-nitro-L-arginine methyl ester or melatonin prevented the reduction of AChE activity caused by acute Pro administration, suggesting the participation of oxidative stress in such effects. PMID- 15789143 TI - How urgent is urgency? A review of current methods of assessment. PMID- 15789142 TI - Acute cholecystitis at initial presentation of polyarteritis nodosa. AB - Polyarteritis nodosa (PAN) is a systemic vasculitis of unknown etiology. Although gastrointestinal involvement may be seen in half of the cases of PAN, vasculitis of the gallbladder at the disease onset is a rare presentation. We report a case of a 48-year-old man who was admitted due to acute cholecystitis. He had complained of myalgia, fever and weight loss for about one month prior to admission. At physical examination, mild hypertension and calf pain were noted. He underwent a cholecystectomy; histopathological evaluation disclosed necrotizing vasculitis suggestive of PAN. We emphasize that cholecystitis may be part of the initial presentation of systemic vasculitis. PMID- 15789144 TI - Tension-free vaginal tape: a prospective subjective and objective outcome analysis. AB - The purpose of this prospective study was to describe the effects of the tension free vaginal tape (TVT) procedure on subjective and objective outcomes. Eighty seven women (aged 31-95 years) underwent a TVT procedure and were followed for up to 24 months using the Incontinence Impact Questionnaire (IIQ-7), Urogenital Distress Inventory (UDI-6), and a Patient Satisfaction Questionnaire. IIQ-7 scores improved from a mean 51.1 before surgery to 12.7 at 1 month (p<0.001) indicating reduced impact of incontinence on quality of life. UDI-6 scores declined from a mean 61.8 to 21.9 (p<0.001) indicating improvement in urinary symptoms. At 1 month, 91.2% of patients were satisfied. Improvements on all measures were maintained throughout follow-up. Urodynamic evaluation of 57 patients (mean: 15.0 months) showed that 91.2% had a negative stress test. Results indicate significant immediate and sustained improvement in incontinence impact and urinary symptoms, and a high rate of patient satisfaction and objective cure. PMID- 15789146 TI - Prevalence, impact on the quality of life and pathophysiological determinants of nocturia in urinary incontinent women. AB - The objective of this study was to estimate the prevalence of nocturia in incontinent women in a urogynaecologic practice and its association with the quality of life and to estimate the prevalences of the pathophysiological categories among nocturics. From 1 January 2002, all patients with complaints of urinary incontinence were analysed according to a specific protocol: multichannel urodynamic testing, a 1-h International Incontinence Society (ICS) pad test and a 3-day frequency-volume (FV) chart. From 1 June 2002, subjects had to fill in a standardised quality of life questionnaire as well. Nocturia was defined as two or more micturitions per night calculated from the FV chart. Evaluable FV charts were received from 111 patients, and 72 patients completed the questionnaires correctly. The overall prevalence of nocturia was 48.6% (confidence interval: 43.9-53.3%). Nocturia had a negative impact on several aspects of the quality of life. The maximum voided volume was significantly (p=0.005) less in nocturics. The 24-h frequency was higher in nocturics (p=0.001). Nocturics lost more urine during the pad test (p=0.039). The multivariate logistic regression analysis showed that the greater the proportion of 24-h urine excreted at night, the greater the odds of having nocturia and that the lesser the maximum voided volume, the greater the odds of having nocturia. The majority (92.7%) of the nocturics can be classified into one of the responsible pathophysiologic categories: nocturnal polyuria in 51.2%, a low functional bladder capacity in 14.6%, a combination of both in 9.8%, polyuria in 4.9% and a combination of polyuria and nocturnal polyuria in 12.2% of the cases. Nocturia is a frequent symptom among urinary incontinent patients, with a negative impact on several aspects of the quality of life. With a FV chart, nocturics can be classified into one of the responsible pathophysiologic categories in the majority (92.7%) of the cases. PMID- 15789145 TI - Short-term functional and neuroregenerative response of the urethra to ovariectomy and vaginal distension in female rats. AB - The objective of this study was to investigate the effects of ovariectomy (OVX) and vaginal distension (VD) on leak point pressure (LPP) and pudendal nerve regenerative response in the female rat. Twenty rats underwent OVX 3 days prior to either VD or sham distension. Seventeen rats did not receive OVX but underwent either VD or sham distension. Four days after distension, LPP testing was performed. In situ hybridization for beta(II) tubulin mRNA, an indicator of the neuroregenerative response, was performed on motoneurons of the pudendal nerve. In the non-OVX group, LPP was significantly decreased after VD. After OVX, the difference in LPP between VD and sham rats did not quite reach the level of statistical significance. There was a statistically significant interaction between the effects of OVX and VD on LPP. There was no significant difference in in situ hybridization results between any of the groups. No neuroregenerative response of motoneurons of the pudendal nerve was observed after either VD or OVX. PMID- 15789148 TI - Laparoscopic ureteroureteral anastomosis for repair of ureteral injury involving stricture. AB - We describe an approach to laparoscopic ureteroureteral anastomosis for surgical management of ureteral stricture, and review four cases in which this method was used. In these four cases, we observed no intraoperative complications. Patients' length of hospital stay was 1-2 days. Follow-up ranged from 12 to 14 months. All four patients have been asymptomatic, and their follow-up intravenous pyelograms (IVPs) have been normal. This surgical approach warrants further study with larger series and long-term follow-up. PMID- 15789147 TI - Assessing outcome after a modified vaginal wall sling for stress incontinence with intrinsic sphincter deficiency. AB - Forty women with stress incontinence, intrinsic sphincter deficiency (ISD), associated or not with urethral hypermobility, a Valsalva leak point pressure (VLLP)<60 cmH(2)0 and a maximum urethral closure pressure<30 cmH(2)0 underwent in situ vaginal wall sling. The main modification to the technique was the use of two small Marlex meshes placed at the lateral edges of the sling. Outcome was assessed by pad use, surgical results and patients' satisfaction. Data of 39/40 patients were analyzed after a minimum follow-up of 1 year. After surgery 30/39 patients were completely dry (no pads), stress incontinence disappeared in 22/39, and 30/39 patients were satisfied with outcome. Reasons for dissatisfaction included recurrence of stress incontinence in three, infections in one and urge incontinence in five. Overall results are good given this category of patients. The vaginal wall sling can be recommended for patients with ISD because the results are promising, it corrects urethral hypermobility and, in our experience, it does not cause obstruction if correctly performed. PMID- 15789149 TI - Surgical procedures for urethral diverticula in women in the United States, 1979 1997. AB - The objective of this study was to describe national rates of surgery for urethral diverticula in women. Data from the National Hospital Discharge Survey (NHDS), a federal database that samples inpatient hospitals in the United States, were analyzed from 1979 to 1997 for diagnosis and procedure codes using the ICD-9 CM classification system. The difference between the median age-adjusted rates from 1979 to 1988 and 1989 to 1997 were evaluated using the Mann-Whitney U test since the yearly rates fluctuated in a nonlinear fashion. Data from the National Statistics for Ambulatory Surgery (NSAS) database were analyzed from 1994 to 1996 in a similar fashion. The average age of women undergoing surgery for urethral diverticula increased from 41.6+/-15.1 years from 1979 to 1988 to 49.4+/-14.8 years from 1989 to 1997 (p=0.02). The average length of hospital stay decreased from 8.4+/-5.0 days in 1979 to 3.2+/-1.7 days in 1997 (p=0.007). Approximately 27,000 inpatient procedures were performed for the repair of urethral diverticula in the United States over a 19-year period, ranging from an estimated 500 to 3400 cases per year. The median age-adjusted rate of procedures decreased from 14.2 per 1 million women from 1979 to 1988 to 6.4 per 1 million women from 1989 to 1997 (p=0.009). Data from the NSAS indicated that an average of 6.7 surgeries per 1 million women per year were performed in the outpatient setting from 1994 to 1996. Age-adjusted rates of inpatient surgery for urethral diverticula were threefold higher for black compared to white women. Inpatient surgical repair of urethral diverticula is three times as high in black as compared to white women. These procedures are infrequent and rates appear to have decreased over time. PMID- 15789150 TI - Complications of synthetic graft materials used in suburethral sling procedures. AB - Problems relating to the erosion of sling material, through either the vagina or the urethra, have been encountered with almost all kinds of synthetic sling materials. We present four unusual cases of women using different synthetic materials and the complications that occurred. The biopsies were examined histologically and analyzed for collagen and inflammatory reactions. Four patients who underwent suburethral slingplasty previously with different sling materials required surgical management for complications, including one intravesical Ethibond migration, vaginal mucosal mesh erosion in two patients, and one proximal urethral overcorrection with intravesical erosion. We reviewed the literature regarding the amount of mesh erosion and connective tissue reaction with synthetic materials. The efficiency of mesh removal was assessed. The four patients maintained urinary continence after urethrolysis and removal of the mesh. Fibrosis and severe inflammatory reactions were found in the connective tissue adjacent to the mesh as well as the Prolene mesh. Technically, it would be easier to remove the graft of patch sling if rejection or erosion occurs. PMID- 15789151 TI - TVT erosion secondary to a twist in tape. AB - Mesh erosions through vaginal mucosa as well as the urethra following TVT procedures have been reported but are rare. We report a case of a 50-year-old woman who was found to have mesh erosion through vaginal mucosa 8 weeks after her TVT procedure, apparently secondary to a twist in the tape. Excision of approximately 5 mm of tape encompassing the twist was accomplished without difficulty, and allowed for complete healing of the vaginal mucosa and resolution of the patient's pain and irritative voiding symptoms. PMID- 15789153 TI - Evaluating the impact of genetic counseling and testing with signal detection methods. AB - One measure of the impact of genetic counseling and testing (GCT) is the extent to which it fosters behavioral change that is consistent with mutation status. We describe and illustrate how two different signal detection methods, receiver operating characteristic (ROC) analysis and recursive partitioning, can be used in this context to evaluate the impact of GCT. We analyzed real screening behavior data obtained in the 12 months following GCT for Hereditary Nonpolyposis Colon Cancer (HNPCC) using these two different signal detection approaches. Each approach demonstrated that GCT had an impact on behavioral outcomes, and was effective in fostering behavioral outcomes appropriate to mutation status. The ROC approach demonstrated that GCT was effective because mutation positive and mutation negative individuals could be distinguished on the basis of the number of recommended screening behaviors. The recursive partitioning approach demonstrated that GCT was effective because there were generally high rates of adherence to screening guidelines among subjects. The recursive partitioning technique also identified four subgroups of subjects, each with distinct characteristics, for which tailored interventions could be developed to increase rates of adherence to screening guidelines. Signal detection methods are easily implemented and are useful techniques for evaluating the impact of GCT. PMID- 15789154 TI - (Mis)alignments in counseling for Huntington's Disease predictive testing: clients' responses to reflective frames. AB - As a sequel to an earlier paper (Sarangi et al., 2004. J Genet Couns, 13(2), 135 155) examining genetic counselors' initiation of reflective frames, in this paper we analyze the variable ways in which clients respond to such reflective frames in the clinical setting. Of the six types of reflective questions identified, we focus on two types, which recur throughout the counseling protocol: (i) questions about clients' decisions to have genetic testing and (ii) questions exploring the potential impact of a positive or negative test result. The analytic focus here is on the mismatches surrounding clients' apparent readiness to discuss coping with the onset of disease (risk of disease) when they have been asked to discuss coping with genetic test results (risk of knowing). Our theoretical discussion is centered around the notion of alignment as a framework for locating the convergence and divergence of counselors' and clients' agendas in interaction. Drawing on detailed transcripts of 24 Huntington's Disease counseling consultations in South Wales, we analyze 119 counselor-client question-response sequences using the methodology of discourse analysis. Preliminary coding of clients' responses led us to identify three recurrent themes: (a) gaining knowledge as a basis for future action; (b) needing to know as a subjective necessity; and (c) downplaying what can be known. In a further analysis of extended data extracts, we draw attention to how clients display varying degrees of engagement with regard to the testing process and outcomes along the temporal and social axes. At one extreme, clients may take up the opportunity to engage in self-reflection, and thus endorse the legitimacy of the reflective frame. At the other extreme, clients may implicitly or explicitly challenge the relevance of self-reflection, and hence the usefulness of this counselor-initiated routine. We suggest that clients' varied response behaviors result from the perceived need of some clients to display their 'readiness' for predictive testing-an overarching 'meta-question' posed by the very existence of the counseling protocol. PMID- 15789152 TI - Cystic fibrosis prenatal screening in genetic counseling practice: recommendations of the National Society of Genetic Counselors. AB - For over a decade, prenatal screening for cystic fibrosis (CF) has been considered a model for the integration of genetic testing into routine medical practice. Data from pilot studies and public policy discourse have led to recommendations by some professional organizations that CF screening should be offered or made available to pregnant women and their partners, and to couples planning a pregnancy. It is crucial that genetic counselors gain thorough understanding of the complexities of CF and the implications of positive test results, so that they may serve as a reliable, educated referral base and resource for health care providers and their patients. While not all pregnant women will be referred for genetic counseling prior to CF carrier testing, genetic counselors often will be asked to counsel clients after they have a positive test result, or who are found to be at increased risk. Genetic counselors can play an important role in providing accurate and current information as well as support for patients' informed decisions. These recommendations were created by a multicenter working group of genetic counselors with expertise in CF and are based on personal clinical experience, review of pertinent English language medical articles, and reports of expert committees. The recommendations should not be construed as dictating an exclusive course of management, nor does the use of such recommendations guarantee a particular outcome. These recommendations do not displace a health care provider's professional judgment based on the clinical circumstances of a particular client. PMID- 15789155 TI - Differences in individual approaches: communication in the familial breast cancer consultation and the effect on patient outcomes. AB - This multicenter study aimed to assess (i) whether individual clinical geneticists and genetic counselors vary in their communication skills and (ii) whether this variation in communication impacts on patient outcomes, such as anxiety, depression, genetics knowledge, and satisfaction. One hundred and fifty women from high-risk breast cancer families attending their first genetic counseling consultation completed pre and post-consultation self-report questionnaires. The consultations were audiotaped and transcribed verbatim. Univariate analyses showed highly significant differences between individual clinical geneticists and genetic counselors in: facilitating understanding (p < or = 0.001); facilitating active involvement (p < or = 0.001); facilitating partnership building (p = 0.003); addressing emotional concerns (p < or = 0.001); and discussing prophylactic mastectomy (p = 0.017). Multivariate linear regressions showed that this variation in communication resulted in a greater change in patient's depression 4 weeks after the counseling session (p = 0.017). These findings suggest clinical geneticists and genetic counselors have achieved some standardization in communicating information, but showed diversity in their facilitation skills. Communication skills may be a useful area to explore further in this field. PMID- 15789156 TI - The effect of experiential knowledge on construction of risk perception in hereditary breast/ovarian cancer. AB - The purpose of this study was to explore the connection between experiential knowledge of hereditary breast/ovarian cancer and understandings of personal cancer risk. Using a qualitative research design, the investigator conducted in depth interviews with 53 individuals (45 female, 8 male) from families at high risk for hereditary breast/ovarian cancer. Study results showed that two forms of experiential knowledge, empathetic and embodied knowledge, were integral to participants' constructions of their cancer risk. They also illustrated that knowledge derived from experience often took precedence over objective clinical estimates of risk. The paper discusses the clinical implications of these findings and suggests that counseling strategies, which expand upon patient's lived experience and knowledge of the disease, may enhance communication of genetic risk. Assessment of experiential knowledge promises to suggest new ways to frame genetic information that will enable people to better understand their objective risk or to modify exaggerated and/or inaccurate risk perceptions. PMID- 15789157 TI - Consideration of genetic counseling as a career: implications for diversifying the genetic counseling field. AB - Under-representation of racial/ethnic minority counselors has been an ongoing issue in the genetic counseling field. A better understanding of genetic counseling awareness and career consideration may help to increase the number of applicants to genetic counseling training programs from racial/ethnic minorities. This study sampled high school and college students (n = 233) to examine their awareness and perceptions of genetic counseling. Ethnicity, gender, parental level of education, and interest in biology were significant predictors of a subject's genetic counseling awareness; previous awareness of genetic counseling, interest in psychology, and level of education were significant predictors of whether a subject would consider genetic counseling as a career. The findings suggest that knowledge of genetic counseling is lower among racial/ethnic minorities, but that racial/ethnic minorities are just as likely to consider genetic counseling as a career. Awareness of genetic counseling prior to university education may increase racial/ethnic minority representation among potential applicants to genetic counseling training programs. PMID- 15789158 TI - Insurance, acculturation, and health service utilization among Korean-Americans. AB - This study describes the pattern and predictors of ambulatory care utilization among Korean Americans (KAs) living in Los Angeles. Data were gathered via a mail survey. Analysis employed a two-part model: logit model for factors affecting any health care use and truncated negative binomial model for frequency of use given one visit. Use of ambulatory care among KAs was low (2.80 visits during prior 12 months), compared to their counterparts in South Korea and the U.S. population. Variables associated with higher utilization included old age, health needs, and health insurance. Income had a positive effect on health care utilization decisions among the uninsured. Acculturation appeared to be neither a strong nor consistent predictor of ambulatory care utilization among KAs. Of particular concern is the finding that KAs suffer from inadequate access to care due to lack of employment-based health insurance. PMID- 15789159 TI - Health behaviors, chronic disease prevalence and self-rated health of older Asian Indian immigrants in the U.S. AB - The purpose of this study was to examine the correlates of healthy behaviors and self-rated health in middle-aged and older Asian Indian immigrants in the U.S. Asian Indian men (n=162) and women (n=64), 50 years of age or older completed a telephone survey which collected information regarding demographics, behavioral risk factors, acculturation, perceived control, quality of social support, depression, body mass index, chronic disease prevalence, and self-rated health. Participants' average length of residence in the U.S. was 25 years, 52% were normal weight, 41% were vegetarians, 55% incorporated aerobic activity into daily lifestyle, and only 5% smoked. Hypertension and diabetes were most common chronic diseases (31 and 18%, respectively). Younger age, longer length of residence and a bicultural or more American ethnic identity were associated with greater participation in physical activity. Likewise, higher income, a bicultural or more American ethnic identity and depression were associated with higher fat intake. Poor self-rated health was associated with older age, female gender, BMI>25, satisfaction with social support, and greater number of chronic disease conditions. A multitude of factors influence the practice of healthy behaviors and the perceived health of Asian Indian immigrants, which should be addressed when developing culturally appropriate health promotion interventions. PMID- 15789160 TI - South Asian victims of intimate partner violence more likely than non-victims to report sexual health concerns. AB - To assess relationships between intimate partner violence (IPV) and sexual health among South Asian women in Boston. Surveys assessed demographics, IPV and sexual and reproductive health outcomes of women in relationships with men (N=208). In depth interviews explored these issues with women with a history of IPV (N=23). Subjects were majority Indian, non-U.S. citizens, and highly educated. Quantitative data were assessed by logistic regression, qualitative data by a grounded theory approach. About 21.2% of the survey sample reported IPV in the current relationship. These women are 2.6 times as likely to report discolored vaginal discharge in the past year (95% CI=1.27-6.50), 3.1 times as likely to report burning during urination in the past year (95% CI=1.52-6.31) and 3.4 times as likely to report unwanted pregnancy in the current relationship (95% CI=1.33 8.66). Interviewed women described how abuse reduces sexual autonomy, increasing risk for unwanted pregnancy and multiple abortions. Study findings demonstrate the need for increased gynecologic health outreach to abused South Asian women in the U.S. PMID- 15789161 TI - Susceptibility of elderly Asian immigrants to persuasion with respect to participation in research. AB - Familism, respect for authority, and a sense of shame/pride are cultural characteristics that might influence research participation of Asian Americans. We compared 79 elderly Asian immigrants, most of whom immigrated from China or Hong Kong, with 58 elders who were not Asian and mostly not immigrants. Responding to hypothetical situations presented on a self-administered questionnaire, the Asian group professed to be more likely to be influenced by a request from a son/daughter, landlord, physician, or advertisement (p<0.001) and by a monetary incentive (p=0.05). Multivariate adjustment for potential confounders attenuated the strength of these relations, but except in the case of the monetary offer, differences remained statistically significant. Within the Asian group, multivariate logistic regression modeling indicated that years lived in the US was associated with more likelihood of refusing requests to participate in research. We conclude that acculturation or assimilation into American society may build resistance to pressure to participate in research. Our findings also suggest that elderly Asian immigrants may need additional protections to achieve truly informed consent. PMID- 15789162 TI - Paan and Gutka in the United States: an emerging threat. AB - Smokeless tobacco and areca nut usage are integral cultural traditions in South Asia. Paan and gutka are two commonly used products which contain these ingredients. They are immensely popular in South Asia and with South Asian immigrants. Regular paan and gutka use is associated with several deleterious health consequences, most significantly oral cancer. Of particular concern is the markedly increased risk of oral cancer in South Asian immigrants when compared with the natives in new areas of settlement. The South Asian community in the United States is large and rapidly growing. Paan and gutka are legal in the United States, and readily available in ethnic enclaves. Smokeless tobacco prevention and cessation research and interventions have not yet addressed the unique sociocultural circumstances of this growing, at-risk community. The medical, dental, and public health communities need to join forces to combat this emerging threat. PMID- 15789163 TI - Engaging Asian Americans for mental health research: challenges and solutions. AB - Asian American communities have important and unmet mental health needs, but there is comparatively little research data on process and outcomes that can guide evidence-based approaches to mental health care. This paper describes our experience of building research programs in a community-based health care facility, some of the challenges we faced, and barriers that were overcome. We have learned that a) mental health services research can be carried out in a community health center with minimal intrusion on usual patient flow; b) the effort must be shared between the health center and the community; c) barriers to participation in mental health research programs are multifactorial ranging from conceptual, cultural, and attitudinal biases to practical concerns inherent in the ethnic minority population; and d) resistance can be overcome by working with participants' cultural and social needs and using their explanatory belief models when developing and pursuing studies. PMID- 15789164 TI - Prevalence of metabolic syndrome risk factors among young adult Asian Indians. AB - Metabolic syndrome has a high prevalence within the U.S population. Asian Indians have a greater prevalence of the chronic diseases associated with this syndrome compared to Caucasians. This study aimed to determine the prevalence of risk factors of metabolic syndrome in young adult Asian Indians. Behavioral risk factors, dietary intake, and anthropometric measurements were assessed on all study participants (n=50). The mean BMI was 23.2 and 20.4, waist circumference was 87 and 79 cm, and percent body fat was 16 and 26% for males and females, respectively. Macronutrient contributions to the total energy intake were: carbohydrate 55% for males and females, protein 14 and 12% for males and females respectively, and total fat 31 and 33% for males and females, respectively. Using the definition of the Third Report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III, ATP III), these Asian Indians did not appear to be at high risk for developing metabolic syndrome. However, using the newly proposed recommendations for Asian Indians, the results suggest that this group may be at risk for developing metabolic syndrome. PMID- 15789166 TI - Computational model of touch sensory cells (T Cells) of the leech: role of the afterhyperpolarization (AHP) in activity-dependent conduction failure. AB - Bursts of spikes in T cells produce an AHP, which results from activation of a Na+/K+ pump and a Ca2+-dependent K+ current. Activity-dependent increases in the AHP are believed to induce conduction block of spikes in several regions of the neuron, which in turn, may decrease presynaptic invasion of spikes and thereby decrease transmitter release. To explore this possibility, we used the neurosimulator SNNAP to develop a multi-compartmental model of the T cell. The model incorporated empirical data that describe the geometry of the cell and activity-dependent changes of the AHP. Simulations indicated that at some branching points, activity-dependent increases of the AHP reduced the number of spikes transmitted from the minor receptive fields to the soma and beyond. More importantly, simulations also suggest that the AHP could modulate, under some circumstances, transmission from the soma to the synaptic terminals, suggesting that the AHP can regulate spike conduction within the presynaptic arborizations of the cell and could in principle contribute to the synaptic depression that is correlated with increases in the AHP. PMID- 15789165 TI - Validity of self-reported weight and height: comparison between immigrant and non immigrant Mexican Americans in NHANES III. AB - This study examined whether 1) the immigrant Mexican Americans reported weight and height differently from the non-immigrant Mexican Americans and, 2) self reporting errors influenced weight classification. A U.S. national sample of 3085 Mexican Americans from the third National Health and Nutrition Examination Survey was used to investigate validity using paired t-tests, multiple linear regressions, sensitivity and specificity. While immigrant Mexican American women reported their weight and height higher than non-immigrant women, no significant differences were found between immigrant and non-immigrant Mexican American men. Sensitivity and specificity of weight classification with self-reported weight and height were not different between immigrant and non-immigrant Mexican Americans; however, underweight Mexican Americans showed the lowest sensitivity. Therefore, self-reported weight and height can be used for the purpose of weight classification with immigrant and non-immigrant Mexican Americans, with an exception of underweight Mexican Americans. PMID- 15789167 TI - Spike timing-dependent synaptic plasticity in visual cortex: a modeling study. AB - Recent studies show that synaptic modification depends critically on the relative spike timing of pre- and postsynaptic neurons. Here we explore the functional implications of spike timing-dependent synaptic plasticity in the visual cortex using a model circuit with modifiable intracortical excitatory connections. First we simulated the experiments using two-point stimuli, in which two visual stimuli in a topographically represented feature space were repeatedly presented in quick succession, and found that tuning of the cortical neurons was modified in a manner similar to that observed experimentally. We then explored the dependence of results on the model parameter and identified the intracortical parameters that were critical for the magnitude of the shifts and obtained a simple relationship between the amount of shift and (S=(sigmaEXTCrec_exc)/sigmaINHCrec_inh). Finally we investigated the effects of moving stimuli in a topographically represented visual space and found that they can effectively induce spike timing-dependent modification of the intracortical connections. It suggests the importance of moving stimuli in dynamic modification of the cortical maps through spike timing-dependent synaptic plasticity. PMID- 15789168 TI - A model for representing the dynamics of a system of synfire chains. AB - Competitive synchronization among synfire chains may model the dynamics of binding and compositionality. Typically, such models require simulations of hundreds of thousands of neurons. Here we show that the behavior of such large systems can be numerically analyzed by representing the neuronal activity in a synfire chain as a wave. The position and velocity of waves are the only parameters needed to represent the neural activity within a synfire chain. With this wave model we describe how waves are generated, decay, interact within a single chain and among chains. The behavior of the wave model is compared to the behavior of detailed simulations of synfire chains with no qualitative difference. We show that interacting waves tend to become locked to each other (wave synchronization). Finally we prove that: (1) Within a system of many synfire chains with symmetric interchain connections, as long as waves do not fade away or become fully synchronized, the total synchrony among waves can only increase (or stay constant), but never decrease. (2) A wave that increases its speed during the synchronization process becomes more stable. PMID- 15789169 TI - How can a patient blind to radial motion discriminate shifts in the center-of motion? AB - Within biologically constrained models of heading and complex motion processing, localization of the center-of-motion (COM) is typically an implicit property arising from the precise computation of radial motion direction associated with an observer's forward self-motion. In the work presented here we report psychophysical data from a motion-impaired stroke patient, GZ, whose pattern of visual motion deficits is inconsistent with this view. We show that while GZ is able to discriminate direction in circular motions she is unable to discriminate direction in radial motion patterns. GZ's inability to discriminate radial motion is in stark contrast with her ability to localize the COM in such stimuli and suggests that recovery of the COM does not necessarily require an explicit representation of radial motion direction. We propose that this dichotomy can be explained by a circular template mechanism that minimizes a global motion error relative to the visual motion input, and we demonstrate that a sparse population of such templates is computationally sufficient to account for human psychophysical performance in general and in particular, explains GZ's performance. Recent re-analysis of the predicted receptive field structures in several existing heading models provides additional support for this type of circular template mechanism and suggests the human visual system may have available circular motion mechanisms for heading estimation. PMID- 15789170 TI - Spectral analysis of event-related hemodynamic responses in functional near infrared spectroscopy. AB - The goal of this paper is to design experiments that confirm the evidence of cognitive responses in functional near infrared spectroscopy and to establish relevant spectral subbands. Hemodynamic responses of brain during single-event trials in an odd-ball experiment are measured by functional near infrared spectroscopy method. The frequency axis is partitioned into subbands by clustering the time-frequency power spectrum profiles of the brain responses. The predominant subbands are observed to confine the 0-30 mHz, 30-60 mHz, and 60-330 mHz ranges. We identify the group of subbands that shows strong evidence of protocol-induced periodicity as well as the bands where good correlation with an assumed hemodynamic response models is found. PMID- 15789171 TI - Neuronal mechanisms encoding global-to-fine information in inferior-temporal cortex. AB - Sugase et al. found that global information is represented at the initial transient firing of a single face-responsive neuron in inferior-temporal (IT) cortex, and that finer information is represented at the subsequent sustained firing. A feed-forward model and an attractor network are conceivable models to reproduce this dynamics. The attractor network, specifically an associative memory model, is employed to elucidate the neuronal mechanisms producing the dynamics. The results obtained by computer simulations show that a state of neuronal population initially approaches to a mean state of similar memory patterns, and that it finally converges to a memory pattern. This dynamics qualitatively coincides with that of face-responsive neurons. The dynamics of a single neuron in the model also coincides with that of a single face-responsive neuron. Furthermore, we propose two physiological experiments and predict the results from our model. Both predicted results are not explainable by the feed forward model. Therefore, if the results obtained by actual physiological experiments coincide with our predicted results, the attractor network might be the neuronal mechanisms producing the dynamics of face-responsive neurons. PMID- 15789172 TI - A cellular mechanism for graded persistent activity in a model neuron and its implications in working memory. AB - Working memory represents the ability of the brain to hold information for relatively short periods of time. Working memory is believed mediated by persistent neuronal firing. A broadly accepted hypothesis is that persistent activity is generated by reverberating synaptic input. However, single cortical neurons capable of showing persistent firing were recently reported. In this modeling study, we propose a cellular mechanism to generate persistent firing of multiple firing rates in single neurons. In the proposed mechanism, bistable concentrations of inositol 1,4,5-trisphosphate (IP3) and Ca2+ is achieved by IP3 formation and IP3-induced Ca2+ release from stores in multiple subcellular domains. A postsynaptic firing rate-dependent switching of these bistable elements can demonstrate graded persistent firing of the rat entorhinal neurons. Such a firing rate-dependent switching may be extended to a variety of intracellular Ca2+ signaling cascades. PMID- 15789173 TI - Comparative pharmacodynamic evaluation of eptifibatide and abciximab in patients with non-ST-segment elevation acute coronary syndromes: the TAM2 study. AB - BACKGROUND: The importance of the relationship between clinical outcome and degree of platelet aggregation inhibition (PAI) achieved with the dosing regimens of GPIIb-IIIa inhibitors used in large trials in patients with non-ST segment elevation (NSTE) acute coronary syndromes (ACS) is increasingly appreciated. In the PURSUIT trial, eptifibatide treatment that consistently provided >80% PAI was associated with clinical benefit at 30 days and 6 months. The GUSTO IV ACS trial, however, did not show any effect of abciximab on 30-day outcomes. This difference might be due to variability of antiplatelet effects of these drugs. As previous studies found, a 12 hr abciximab infusion had <80% PAI, particularly at 6 and 12 hr. These studies did not evaluate PAI with a longer, 24-hour infusion as used in GUSTO IV ACS. METHODS: We conducted a prospective study in 40 patients with NSTE ACS prior to catheterization or coronary intervention at 3 centers using the PURSUIT dose of eptifibatide (180/2.0) [DOSAGE ERROR CORRECTED] and the GUSTO IV dose of abciximab (0.25, 0.125). Blood samples were collected at baseline, and during the infusion at 10 min, 1 hr, 6 hr, 8 hr, 12 hr, 18 hr, and 24 hr. Measurements of ex vivo light transmission aggregometry (LTA) were performed using PPACK anticoagulant and 20 microM ADP agonist. Receptor Occupancy (RO) was also determined in a subset of patients. RESULTS: Eptifibatide achieves higher PAI during the entire infusion period than abciximab (p<0.01). At 10 min, average PAI with eptifibatide and abciximab was 88% and 80%, respectively, 95% and 79% at 6 hr, and 97% and 79% at 24 hr. There was also more variability in individual patient response to abciximab. Although average RO for eptifibatide was similar to that of abciximab at 10 min, 67% versus 69%, respectively, average RO was higher in the eptifibatide cohort at all subsequent timepoints. By 24 hr, average RO for eptifibatide was 86%, whereas abciximab averaged 67%. CONCLUSION: These data support the hypothesis that differences in clinical outcomes of large GPIIb IIIa trials in patients with NSTE ACS may be related to the consistency and potency of antiplatelet effects of the dosing regimens used. PMID- 15789174 TI - Metabolism of homocysteine and its relationship with cardiovascular disease. AB - Hyperhomocysteinemia, or the rise of plasmatic homocysteine levels above 15 mug/dL, is accepted nowadays as an independent risk factor for cardiovascular disease in men and women. Homocysteine (Hcy) is a non-protein forming aminoacid (aa) derivated from the loss of the methyl group, found within methionine. Methionine regenerates by retrieving the methyl radical from 5 methyltetrahydrofolate (5-MTHF) creating tetrahydrofolate (THF) which will then regenerate to 5-MTHF through the action of methylentetrahydrofolate reductase (MTHFR). This process is called remethylation. Alternatively, Hcy can follow the transsulfuration route, where through cystationine-beta-syntetase (CBS), it irreversibly converted into cystationine, a precursor of cysteine, glutathione, and other substances that are finally excreted in the urine. Hyperhomocysteinemia results from inhibition of the remethylation route, or inhibition or saturation of the transsulfuration pathway. Main factors causally associated increased plasmatic Hcy are mutations of the enzymes MTHFR and CBS; varying nutritional and health states; demographic factors; and, others. The most accepted hypotheses about Hcy action in cardiovascular disease are direct endothelial and vessel wall damage; oxidative stress generation; and, stimulation of a procoagulant and proinflammatory state of blood components. Since hyperhomocysteinemia can be effectively treated with folic acid, prospective trials are underway to determine if folate therapy is required to lower Hcy levels in plasma. These studies also attempt to address the impact, if any, of folate therapy in the reduction of cardiovascular risk, and to demonstrate if hyperhomocysteinemia is actually an independent risk factor that can be effectively treated. PMID- 15789175 TI - Pathological changes in acute coronary syndromes: the role of statin therapy in the modulation of inflammation, endothelial function and coagulation. AB - Considerable progress has been made in our understanding of the pathophysiology of coronary artery disease (CAD), their acute presentations as acute coronary syndromes (ACS) and the role of LDL cholesterol. In particular there is clear evidence that atherosclerosis is far from being a process that leads to an amorphous flow limiting lesion on an angiogram, but rather involves a complex interplay between the endothelium, inflammatory cells and the coagulation cascade occurring throughout the coronary vascular bed. While a culprit flow limiting lesion may be effectively treated by a drug eluting stent or coronary bypass surgery, this will have little impact on the global molecular processes that determine recurrent plaque instability at non-culprit sites. The search for systemic long term therapy, which is safe and effective and reduces the changes in inflammation, endothelial function and thrombosis that are the hallmark of ACS, has pushed statins to the forefront. A number of recent clinical trials have shown the benefits of early statin therapy in the treatment of ACS. In addition to their effects on LDL cholesterol, statins have a number of properties collectively referred to as pleiotropic effects, which enable them to modulate the adverse biological changes that are associated with ACS. The purpose of this review is to acquaint the reader with the biological changes that accompany ACS, highlight how these pathways may be modulated for clinical benefit by statins and identify potential novel targets for future therapy. PMID- 15789176 TI - Analytical performance of the new coagulation monitoring system INRatio for the determination of INR compared with the coagulation monitor Coaguchek S and an established laboratory method. AB - An evaluation of the INRatio Prothrombin Time Monitoring system for determination of INR was done in two centers with a total of 5 healthy subjects and 77 subjects on oral anticoagulation. The INRatio and the Coaguchek S were compared with an established laboratory method. The correlation coefficient of the comparison with the laboratory was r=0.954 for INRatio and r=0.937 for Coaguchek S. The mean relative deviation from the lab method calculated according to Hill was 6.87% for INRatio, which is rated "very goo", and 9.72% for Coaguchek S ("goo"). The imprecision in the normal range (INR=1.1) showed a coefficient of variation (CV) of 7.8% and a standard deviation (SD) of 0.09. In the therapeutic range (INR 3.9) the CV was 5.4%, the SD 0.21 and above therapeutic range (INR 5.3), the CV was 8.4% (SD 0.44), rated satisfactory. The concordances of the compared methods with the routine were 81% for INRatio and 79% for Coaguchek S, which is considered state-of-the-art. Most of the patients' perceptions of the INRatio were very positive. In the hands of professionals the INRatio demonstrated very good accuracy and precision and an excellent technical reliability. Further studies using INRatio for self testing by patients are warranted. PMID- 15789177 TI - Point-of-care determination of baseline platelet function as a predictor of clinical outcomes in patients who present to the emergency department with chest pain. AB - BACKGROUND: Increased platelet reactivity can identify patients at high risk for thrombotic events, but its clinical use has been impractical due to technical limitations. The purpose of the present study is to determine if a point-of-care measurement of platelet function in patients presenting to an emergency room with chest pain can identify those at high risk of adverse cardiac events. METHODS: Platelet function was measured using the Ultegra-RPFA in 201 patients presenting to the emergency department with the primary complaint of chest pain and either known coronary disease or at least one cardiac risk factor. The primary endpoint was any major adverse cardiac events (MACE) [cardiac death, myocardial infarction (MI), re-admission for coronary revascularization] 6 months post-enrollment. RESULTS: Platelet function at baseline ranged from 44 to 315 platelet activation units (PAU) (mean 175+/-6). Seventy-six patients experienced MACE (37.8%) by 6 months post-enrollment. Mean PAU was significantly lower in the group experiencing MACE (166+/-9 vs. 181+/-9; p=0.026). By univariate analysis, admission PAU was a significant predictor of MACE at 6 months (p=0.028). However, when adjusted for age, gender, cardiac risk factors, and a history of coronary artery disease (CAD) using multivariate logistic regression analysis, PAU was no longer significantly predictive of MACE (p=0.268). CONCLUSIONS: Point-of-care testing of platelet function deserves further study for risk assessment and individualized therapy in the future. PMID- 15789178 TI - Pulmonary embolism in medical patients: improved diagnosis and the role of low molecular-weight heparin in prevention and treatment. AB - Pulmonary embolism is the major complication of deep-vein thrombosis (DVT) and has been shown in autopsy studies to account for 5-10% of hospital deaths. Approximately 75% of fatal pulmonary emboli (PE) occur in medical patients, a clinically heterogeneous group of patients. Ten percent of deaths due to PE occur within the first hour of the acute thrombotic event. Unfortunately, only 25% of all PE are diagnosed and receive appropriate treatment. The major focus for preventing this complication in medically ill hospitalized patients is to provide venous thromboembolism (VTE) prophylaxis. Unfractionated heparin (UFH) was the mainstay for preventing VTE in hospitalized medical patients. However, over the past few years, low-molecular-weight heparins (LMWHs) have been shown to be at least as effective and safer in preventing VTE in this population. Although highly effective in preventing VTE, there remains a small incidence of thrombotic events. Therefore, clinicians must be attentive to the symptoms and signs that would indicate a PE. If a PE is confirmed, then appropriate treatment based on the clinical status of the patient with either LMWH or body-weight dose-adjusted UFH is indicated. PMID- 15789179 TI - Self-management of oral anticoagulant therapy: a review. AB - Patient self-management of oral anticoagulant therapy has over the last years gained increasing attention and widespread use. The method implies self-testing using a coagulometer (point-of-care device) and self-dosaging of coumarin. The method entails advantages for selected patients, who do not need to go to the hospital or family doctor for blood specimen and drug dosage adjustment. In contrast to patients on conventional management, patients performing self managing have a reduced binding to the health care system, and it allows these patients unhindered to travel and manage their job or school. Therefore, the patients have a high degree of compliance because they are highly motivated. This has been the main justification for patient self-management. Furthermore, the conducted trials have also indicated a better quality of treatment compared to conventional management. In this paper the different aspects of this new treatment modality are critically reviewed, with special focus on the published clinical studies and the evaluation of the used coagulometers. PMID- 15789181 TI - Genetic polymorphisms and thromoembolic risk in atrial fibrillation. PMID- 15789180 TI - Successful lysis of intra-cardiac thrombi with streptokinase in patients with renal failure; two case reports and review of the literature. AB - In situ formation of thrombi within the cardiac cavities carries a substantial risk of morbidity and mortality due to the inherent danger of embolization to vital organs. This typically occurs in patients with underlying cardiac disorders associated with low flow state favoring regional stagnation of blood and subsequent clotting. Occasionally, extra-cardiac conditions such as renal failure predispose to the development of intra-cardiac thrombi in the presence or absence of identifiable cardiac abnormalities. Once identified, the appropriate treatment of such thrombi remains highly controversial. Here, we report on the successful and safe use of streptokinase in two patients with end stage renal disease with high risk left ventricular, right ventricular and right atrial thrombi. PMID- 15789182 TI - Factor Xa-protease activated receptor interface: a paradigm for combination pharmacotherapy. PMID- 15789183 TI - Treating problem gamblers: working towards empirically supported treatment. AB - There is an urgent need for mental health professionals to develop effective treatments for pathological gambling. A scientific meeting was held in Quebec City in June 2002 where 13 specialists in the treatment of gamblers pursued the objectives of making the "State of the art" statement concerning the procedures and the methodology to be used in future treatment outcome studies. This paper introduces this special issue of the Journal of Gambling Studies and identifies some of the key issues that confront clinicians and researchers interested in evidence based treatment for gambling related disorders. PMID- 15789184 TI - Conceptual and methodological issues in treatment outcome research. AB - Few studies have evaluated the comparative efficacy, effectiveness and mode of action of specific treatment interventions, or developed evidence-based best practice management guidelines for the treatment of problem gambling. While individual and multi-modal programmes demonstrate positive results, systematic well-designed randomised controlled studies incorporating follow-up blind assessments and standardised diagnostic and outcome measures are needed to establish long-term outcomes. PMID- 15789185 TI - Implications of a brief intervention trial for problem gambling for future outcome research. AB - We describe the development of an empirically supported brief workbook intervention. The implications for the design of future outcome research include the use of media recruitment of participants, the value of a broad conceptual treatment model, the limitations of a waiting list control design, the need for consensus on the measurement of outcome indicators. PMID- 15789186 TI - Psychological treatment of slot-machine pathological gambling: new perspectives. AB - In this paper the most relevant findings of our research team on pathological gambling in the last decade are presented. There is no conclusive empirical evidence of a specific profile of the pathological gamblers. The choice treatment appears to be stimulus control and in vivo exposure with response prevention, followed by a cognitive-behavioral intervention in relapse prevention. Predictive variables for the therapeutic failure were the dissatisfaction with the treatment, the alcohol abuse and the neuroticism as a personality variable. Unanswered questions for future research in this field are commented upon. PMID- 15789187 TI - Gamblers anonymous and cognitive-behavioral therapies for pathological gamblers. AB - Numerous types of treatments for pathological gambling have been described, but two of the most common are Gamblers Anonymous (GA) and cognitive-behavioral therapy. This paper describes some outcome data associated with the two approaches. It also reviews evidence suggesting that a combined intervention may enhance therapy engagement and reduce relapse rates. PMID- 15789188 TI - Evaluation and treatment of pathological gambling. AB - The aim of this article was to describe a model for evaluating and implementing cognitive-behavioral treatment for pathological gambling. The model takes into account the fact that pathological gamblers form a heterogeneous group with varied biopsychosocial characteristics. PMID- 15789189 TI - Pathological gambling, comorbidities and treatment outcome: is the bronze standard good enough? A case study. AB - The paper presents a case study of our attempt to design an outcome package for a program including pathological gamblers and their comorbidities. The clinical intricacies involved highlight the limitations of the clinical trial and the need for alternate designs aimed at assessing effectiveness. PMID- 15789190 TI - Controlled gambling for pathological gamblers. AB - Despite its high prevalence, pathological gambling often remains untreated. It is estimated that only 10% of the pathological gamblers identified in prevalence studies will enter treatment. Within this small proportion, a high percentage will drop out. Despite the facts that some researchers argue against abstinence as the unique treatment goal and that regaining control appears to be possible for some pathological gamblers, abstinence has been the only treatment goal in most problem gambling interventions thus far. This paper examines the avenue of controlled gambling embedded in a harm reduction context as a viable solution for some pathological gamblers. PMID- 15789191 TI - The Iowa Gambling Treatment Program: treatment outcomes for a follow-up sample. AB - The Iowa Gambling Treatment Program (IGTP) amassed participant data for gamblers and concerned others of gamblers over 4 years (1997-2001). Data collection opportunities included: (1) crisis contacts, (2) placement screening, (3) admission, (4) treatment services, (5) discharge, and (6) follow-up. Among followed gamblers, 74% of treatment completers, 49% of substantial treatment completers, and 36% of dropouts and referrals were abstaining from gambling at 6 month follow-up. The reduction in dollars lost to gambling was similar; 85% of treatment completers, 88% of substantial treatment completers, and 65% of others reduced their dollars lost per week. Although more extensive follow-up efforts are needed to properly evaluate the effectiveness of the IGTP, these results suggest that the IGTP shows promise. PMID- 15789192 TI - A perspective on problem gambling treatment: issues and challenges. AB - Researchers face several challenges in conducting gambling treatment research. These include the impact of monetary incentives to participate, difficulty in subject recruitment, treatment ambivalence, heterogeneity of gambling behaviors among treatment samples, the role of natural recovery, the impact of intractable financial pressures, and the specification of adequate process and outcome measures. Each challenge is defined and potential resolutions suggested. PMID- 15789193 TI - Problems in measuring the effectiveness of cognitive therapy for pathological gambling. AB - Cognitive therapy is a relatively new approach to the treatment of pathological gambling. Theoretically, there are strong grounds for believing that cognitive treatments will be effective in helping individuals cut back and stop excessive levels of gambling. However, there is evidence that cognitive therapy for pathological gambling is being confused with cognitive-behaviour therapy. In this paper, the distinction between treatments that are cognitive and those that are cognitive-behavioural is highlighted. Such a distinction has strong implications for the manualisation of therapy. Additionally, a range of problems that confront the evaluation of all therapies for pathological gambling is considered. Spontaneous recovery without therapeutic intervention has been documented in both field studies of both problem and non-problem players and controlled trials of cognitive therapy compared to a waiting list control group. The implications of the phenomenon of spontaneous recovery for the evaluation of cognitive therapy are described. Other problems common to all evaluations of psychotherapies are considered in relation to gambling and recommendations made for outcome study designs. PMID- 15789194 TI - Advancing treatment strategies for pathological gambling. AB - Experiences from treatment trials are described. The use of structured instruments for subject characterization and symptom change assessment is discussed and the challenges of treating specific groups (e.g., those with co occurring alcohol use disorders) explored. Treatment for pathological gambling might be advanced most rapidly through collaborative multi-center efforts. PMID- 15789196 TI - Methodological problems in research on treatments for pathological gambling. AB - These common methodological problems affect the usefulness of pathological gambling (PG) treatment research: (1) Most PG etiological models derive from substance abuse; (2) Most PGs recover on their own and few seek treatment, so PGs in treatment studies may not be representative; (3) Data on treatment-seeking PG women, older adults, and minority group members, or on group, marital, and psychodynamic approaches to PG, are sparse; (4) Little research has examined the impact of comorbidity on PG treatment response; (5) Design deficiencies burden many PG treatment studies. PMID- 15789195 TI - Pharmacological treatments of pathological gambling. AB - Medication treatment studies have demonstrated short-term efficacy of various SRIs, opioid antagonists, and mood stabilizers in sub-samples of adult treatment seeking pathological gamblers. Pathological gambling is frequently comorbid with bipolar spectrum disorders, substance abuse/dependence, and attention deficit/hyperactivity disorder (ADHD), and comorbidity may influence treatment response in pathological gambling. This review focuses on recent research examining the treatment of pathological gambling and highlights methodological challenges for future studies. PMID- 15789197 TI - [Dyskeratosis congenita in a 40-year-old patient]. AB - A 40-year-old patient with a 3-year history of thrombocytopenia was admitted with reticulated and speckled hyper- and hypopigmentations especially on the upper trunk. Aplasia or dystrophy of the fingernails and toenails as well as atresia of the lacrimal ducts were noted. Examination of the oropharynx revealed multiple mucosal leukoplakias and loss of almost all teeth. Based on these observations the diagnosis of X-linked dyskeratosis congenita (Zinsser-Cole-Engman syndrome, OMIM #305000) was made and confirmed by sequencing of the dyskerin 1 (DKC1) gene which revealed a missense mutation in exon 11. PMID- 15789198 TI - [Established and current procedures in wound healing]. AB - Most chronic wounds are caused by arterial or venous vascular disease. Wound care is an interdisciplinary task and economic challenge. Numerous new wound dressings and treatment methods have been introduced recently. Basic research has enhanced our understanding of stimulation and inhibition of wound healing. Well constructed clinical studies have shown some traditional approaches to be effective and others, less so. Successful wound healing requires treatment of the underlying disease as well as correction of local factors that may delay healing. The choice of dressings must be based on continuous re-assessment of the wound. Modern approaches for the most common types of chronic wounds, as well as options such as vacuum treatment and tissue-engineered skin are presented along with information on latest rules for reimbursement for wound care in Germany. PMID- 15789199 TI - [Allergic contact dermatitis from a hydrocolloid dressing due to colophony sensitization]. AB - A 62-year-old female patient with a venous leg ulcer developed massive eczema during wound bed preparation with the hydrocolloid dressing Varihesive. The patch testing confirmed a pronounced sensitization to the hydrocolloid dressing apart from the sensitization to colophony. After review of the current literature we found several case reports from the last 10 years about sensitization to hydrocolloids which were identical but distributed under different brand names in different countries. These dressings contain the pentaerythritol ester of hydrogenated rosin as the tackifying agent which is the substance retaining the sensitizing potential of colophony. Especially patients with chronic wounds frequently tend to contact sensitizations, and colophony currently represents the 4th most frequent allergen in Germany. Therefore, highly potent allergens such as colophony should be strictly avoided as a content material of modern wound dressings. PMID- 15789200 TI - Diagnosis of middle cerebral artery occlusive lesions with contrast-enhanced transcranial color-coded real-time sonography in acute stroke. AB - It is useful to evaluate the occlusive lesions of middle cerebral artery (MCA) occlusion with transcranial color-coded real-time sonography (TCCS). However, TCCS criteria for locating the site of the MCA occlusion has, as yet, remained unclear. The aim of the present study was to establish TCCS criteria for MCA occlusive lesions. We prospectively performed contrast-enhanced TCCS (CE-TCCS) in 75 consecutive acute stroke patients within 24 h of digital subtraction angiography. Patients were divided into four groups: occlusion of the MCA stem (MO group, n=12); occlusion of the MCA branch (MBO group, n=10); stenosis of the MCA stem (MS group, n=9); and no occlusive or stenotic lesions (control group, n=44). The following parameters were measured: peak systolic velocity (PSV) and end diastolic velocity (EDV) of bilateral MCA stems, and ED-ratio (the side-to side ratio of the EDV). We establish the CE-TCCS criteria for MCA occlusive lesions using the sensitivity-specificity curve analysis. A PSV of 170 cm/s distinguished MCA stenosis from other groups (positive and negative predictive values and the accuracies were 100%, 99.0% and 99.1%, respectively). An EDV of 26 cm/s differentiated MO or MBO from the other groups (positive and negative predictive values and the accuracies were 84.6%, 100% and 96.5%, respectively). An ED-ratio of 2.5 discriminated MO from the MBO group (positive and negative predictive values and the accuracies were 88.9%, 85.7% and 87.5%, respectively). Measurement of MCA stem flow velocity with CE-TCCS can identify MCA stem stenosis and occlusion, as well as MCA branch occlusion. PMID- 15789201 TI - Life-threatening bleeding from a vertebral artery pseudoaneurysm after anterior cervical spine approach: endovascular repair by a triple stent-in-stent method. Case report. AB - The incidence of injury to the cervical vertebral artery during surgery for stenosis of the cervical neuroforamina is very low. We present a case in which bleeding during microforaminotomy at the level C6/7 occurred. The bleeding could be controlled intraoperatively. Two days later, a life-threatening cervical hematoma required urgent bedside evacuation. A false aneurysm of the left cervical vertebral artery was successfully occluded by a modified triple stent-in stent technique, maintaining the flow in the vessel. PMID- 15789202 TI - Clinically silent choroid plexus cyst: evaluation by diffusion-weighted MRI. AB - We retrospectively reviewed diffusion-weighted magnetic resonance images of 57 patients with a choroid plexus cyst diagnosed by contrast-enhanced T1-weighted imaging. All the cysts appeared to represent incidental findings. Thirty-eight of 57 patients had bilateral cysts and 19 had unilateral ones. On diffusion-weighted images, 78 of 95 cysts showed homogeneously high signal intensity, 12 showed focal high signal areas, and 5 had no portion with a high signal. The apparent diffusion coefficient of the high signal areas in the cysts was (1.46+/-0.14) x10(-3) mm(2)/s, intermediate between the apparent diffusion coefficients of cerebrospinal fluid and cerebral white matter, (3.15+/-0.67) x10(-3) and (0.79+/ 0.22) x10(-3) mm(2)/s, respectively. Pathological correlation was available in one case, showing high signal intensity areas in the glomera of the choroid plexuses in the lateral ventricles on diffusion-weighted images corresponding to gelatinous cysts with highly proteinaceous content. PMID- 15789203 TI - MRI findings in an atypical case of Kearns-Sayre syndrome: a case report. AB - MR imaging features of mitochondrial encephalomyopathies, lactic acidosis, and stroke-like episodes, Kearns-Sayre/Pearson syndrome have been described in the literature. We describe extensive white matter changes with abnormal signal intensity lesions involving the deep gray nuclei and myelinated white matter tracts in an 18-year-old female with a large-scale 7.4 kb mitochondrial DNA deletion and a atypical presentation of Kearns-Sayre syndrome. Restricted diffusion due to status spongiosus at the involved sites is also discussed. PMID- 15789204 TI - Purification and characterization of NAD(P)H-dependent nitroreductase I from Klebsiella sp. C1 and enzymatic transformation of 2,4,6-trinitrotoluene. AB - Three NAD(P)H-dependent nitroreductases that can transform 2,4,6-trinitrotoluene (TNT) by two reduction pathways were detected in Klebsiella sp. C1. Among these enzymes, the protein with the highest reduction activity of TNT (nitroreductase I) was purified to homogeneity using ion-exchange, hydrophobic interaction, and size exclusion chromatographies. Nitroreductase I has a molecular mass of 27 kDa as determined by SDS-PAGE, and exhibits a broad pH optimum between 5.5 and 6.5, with a temperature optimum of 30-40 degrees C. Flavin mononucleotide is most likely the natural flavin cofactor of this enzyme. The N-terminal amino acid sequence of this enzyme does not show a high degree of sequence similarity with nitroreductases from other enteric bacteria. This enzyme catalyzed the two electron reduction of several nitroaromatic compounds with very high specific activities of NADPH oxidation. In the enzymatic transformation of TNT, 2-amino 4,6-dinitrotoluene and 2,2',6,6'-tetranitro-4,4'-azoxytoluene were detected as transformation products. Although this bacterium utilizes the direct ring reduction and subsequent denitration pathway together with a nitro group reduction pathway, metabolites in direct ring reduction of TNT could not easily be detected. Unlike other nitroreductases, nitroreductase I was able to transform hydroxylaminodinitrotoluenes (HADNT) into aminodinitrotoluenes (ADNT), and could reduce ortho isomers (2-HADNT and 2-ADNT) more easily than their para isomers (4 HADNT and 4-ADNT). Only the nitro group in the ortho position of 2,4-DNT was reduced to produce 2-hydroxylamino-4-nitrotoluene by nitroreductase I; the nitro group in the para position was not reduced. PMID- 15789205 TI - Drought-inducible-but ABA-independent-thaumatin-like protein from carrot (Daucus carota L.). AB - Drought treatment induces the accumulation of dcTLP, which is similar in structure to the thaumatin-like proteins (TLPs) found in the embryogenic calli, seedlings, and mature plants of carrot (Daucus carota). We isolated a full-length dcTLP cDNA clone from carrot and characterized the 5' upstream sequences. The coding region of dcTLP consisted of 645 nucleotides; the theoretical pI value was 4.9, and its molecular weight was approximately 22 kDa. The production of dcTLP transcripts in the seedlings increased dramatically with dehydration treatment but was not affected by abscisic acid (ABA), salicylic acid, or jasmonic acid. The expression patterns of dcTLP mRNA at different developmental stages and in response to a variety of signal molecules was analyzed using reverse transcriptase-PCR and promoter analysis with fused genes of 0.5-kb 5' upstream sequences in which beta-glucuronidase (GUS) reporter genes (gus) were established. The induction of dcTLP was found to be highly specific to drought stress in the embryogenic calli, seedlings, and mature plants. Our results suggest that this new isoform of TLP that has been isolated from carrot is a drought-specific, ABA-independent, non-organ-specific, and non-developmental stage-specific protein. PMID- 15789206 TI - Enhancement of American chestnut somatic seedling production. AB - Somatic embryogenesis holds promise for mass propagation of American chestnut trees bred or genetically engineered for resistance to chestnut blight. However, low germination frequency of chestnut somatic embryos has limited somatic seedling production for this forest tree. We tested the effects of culture regime (semi-solid versus liquid), cold treatment, AC and somatic embryo morphology (i.e., cotyledon number) on germination and conversion of the somatic embryos. Cold treatment for 12 weeks was critical for conversion of chestnut somatic embryos to somatic seedlings, raising conversion frequencies for one line to 47%, compared to 7% with no cold treatment. AC improved germination and conversion frequency for one line to 77% and 59%, respectively, and kept roots from darkening. For two lines that produced embryos with one, two or three-plus cotyledons, cotyledon number did not affect germination or conversion frequency. We also established embryogenic American chestnut suspension cultures and adapted a fractionation/plating system that allowed us to produce populations of relatively synchronous somatic embryos for multiple lines. Embryos derived from suspension cultures of two lines tested had higher conversion frequencies (46% and 48%) than those from cultures maintained on semi-solid medium (7% and 30%). The improvements in manipulation of American chestnut embryogenic cultures described in this study have allowed over a 100-fold increase in somatic seedling production efficiency over what we reported previously and thus constitute a substantial advance toward the application of somatic embryogenesis for mass clonal propagation of the tree. PMID- 15789207 TI - Improved Norway spruce somatic embryo development through the use of abscisic acid combined with activated carbon. AB - The combination of abscisic acid (ABA) and activated carbon increased Norway spruce (Picea abies L., Karst.) cotyledonary somatic embryo yields, increased the number of genotypes forming cotyledonary embryos, caused embryos to form that exhibited improved maturation characteristics, and reduced embryo production costs. Somatic embryos increased in size, showed larger apical regions, became more zygotic-like in shape, and showed higher percentages of epicotyl development upon germination. Analyses of medium for free ABA in the presence of activated charcoal showed a rapid decrease within a few hours followed by a gradual decline over the next few days with little change from 2 to 6 weeks. Gelling agents strongly affected ABA adsorption, with agar decreasing the adsorption of ABA compared to gellan gum (Gelrite, Phytagel). Over 4,000 somatic seedlings from 20 clones were produced and established in a greenhouse using the methods discussed, and approximately 1,250 seedlings representing seven clones were established in a field setting. PMID- 15789208 TI - Agrobacterium tumefaciens-mediated transformation of leek (Allium porrum) and garlic (Allium sativum). AB - Transgenic leek (Allium porrum) and garlic (Allium sativum) plants have been recovered by the selective culturing of immature leek and garlic embryos via Agrobacterium-mediated transformation using a method similar to that described by Eady et al. (Plant Cell Rep 19:376-381, 2000) for onion transformation. This method involved the use of a binary vector containing the m-gfp-ER reporter gene and nptII selectable marker, and followed the protocol developed previously for the transformation of onions with only minor modifications pertaining to the post transformation selection procedure which was simplified to have just a single selection regime. Transgenic cultures were selected for their ability to express the m-gfp-ER reporter gene and grown in the presence of geneticin (20 mg/l). The presence of transgenes in the genome of the plants was confirmed using TAIL-PCR and Southern analysis. This is the first report of leek and "true seed" garlic transformation. It now makes possible the integration of useful agronomic and quality traits into these crops. PMID- 15789209 TI - Comparison of intraparenchymal circulation in various areas of the kidneys of newborns by means of Doppler ultrasound. PMID- 15789210 TI - Imaging strategies in pediatric urinary tract infection. AB - This article is focused on the controversial topic of imaging strategies in pediatric urinary tract infection. A review of the recent literature illustrates the complementary roles of ultrasound, diagnostic radiology and nuclear medicine. The authors stress the key role of ultrasound which has recently been debated. The commonly associated vesicoureteric reflux has to be classified as congenital or secondary due to voiding dysfunction. A series of frequently asked questions are addressed in a second section. The proposed answers are not the product of a consensus but should rather be considered as proposals to enrich the ongoing debate concerning the evaluation of urinary tract infection in children. PMID- 15789212 TI - Protective aprons in imaging departments: manufacturer stated lead equivalence values require validation. AB - The composition of protective aprons worn by X-ray personnel to shield against secondary radiation is changing. Lead is being replaced by either lead-free or composite (lead with other high atomic numbered elements) materials. These newer aprons are categorised by manufacturers in terms of lead equivalent values, but it is unclear how these stated values compare with actual lead equivalent values. In this work, the actual lead equivalence of 41 protective aprons from four manufacturers, all specified as having 0.25 mm lead equivalence, were investigated with transmission experiments at 70 and 100 kVp. All aprons were in current use. The aprons were screened for defects, and age, weight and design was recorded along with details of associated quality assurance (QA). Out of the 41 protective aprons examined for actual lead equivalence, 73% were outside tolerance levels, with actual levels in some aprons demonstrating less than half of the nominal values. The lack of compatibility between actual and nominal lead equivalent values was demonstrated by aprons from three of the four manufacturers investigated. The area of the defects found on screening of the protective aprons were within recommendations. The results highlight the need for acceptancy and ongoing checks of protective aprons to ensure that radiation exposure of imaging personnel is kept to a minimum. PMID- 15789211 TI - Pharmacokinetic analysis of ductal carcinoma in situ of the breast using dynamic MR mammography. AB - The purpose of this study was to assess the relationship between functional parameters derived from dynamic MR imaging and the histological findings of breast ductal carcinoma in situ (DCIS) and DCIS with invasive foci, and to evaluate whether these parameters might predict DCIS patient outcome. Two parameters, amplitude A and k(21), were determined from multicompartmental pharmacokinetic analyses of dynamic MR mammography in 39 patients with needle biopsy-proven primary DCIS. After surgery, the histological tumor characteristics, including microvessel density (MVD) (anti-CD-34), vascular permeability (anti-VEGF antigen) and histological grade, were evaluated. Histology revealed 27 pure DCIS and 12 DCIS with invasive foci. In pure DCIS, positive correlations between MVD and amplitude A (r=0.56, P<0.0025) and between MVD and k(21) (r=0.43, P=0.02) were found. As for histological grade, the differences in both functional parameters of grade 1 versus grade 2 and grade 1 versus grades 2 and 3 combined were significant (P<0.05). No significance was found in the analysis of DCIS with invasive foci. Our results indicated that functional MRI-based parameters might possess the potential to predict the outcome of patients with DCIS. Further study will be needed with larger series over longer periods. PMID- 15789213 TI - Isolated diastasis of cranial sutures: unusual presentation of a blocked shunt in an infant. AB - INTRODUCTION: Ventriculo-peritoneal (VP) shunt malfunction is usually due to blockage of the ventricular catheter and this is typically apparent as enlarged ventricles on a CT scan of the brain. We describe a less common radiological finding in an infant with a blocked shunt. CASE REPORT: A male infant presenting with hydrocephalus in the neonatal period underwent insertion of a VP shunt. He represented at 17 months of age with the clinical features of raised intracranial pressure. A CT scan of the brain revealed that the ventricles were smaller compared with his previous scan, but X-rays taken as part of the shunt series revealed diastasis of the sagittal, coronal and lambdoid sutures. The shunt was explored and the ventricular catheter was found to be blocked and was replaced. Post-operatively there was complete resolution of the symptoms and the suture diastasis. CONCLUSIONS: Suture diastasis with small ventricles on a CT scan of the brain is an unusual radiological finding in an infant with a blocked shunt. Suture diastasis in this patient suggests raised intracranial volume and this may be due to the transependymal absorption of CSF into the white matter. PMID- 15789214 TI - Fetal exposure to sodium valproate associated with Baller-Gerold syndrome: case report and review of the literature. AB - CASE REPORT: We report three patients with a history of maternal valproate use during pregnancy who presented with a combination of metopic suture synostosis and upper limb malformations, which could be diagnosed as Baller-Gerold syndrome (BGS). The patients underwent surgical treatment for the craniofacial deformity, during which standard frontocranial reconstruction was performed. REVIEW OF THE LITERATURE: Only 32 patients have been reported in the world literature and these cases support the emerging view that BGS is not a distinct syndrome, but should instead be considered to be an heterogeneous phenotype with variable etiology. CONCLUSIONS: Our case series suggests for the first time that fetal sodium valproate exposure may also cause this phenotype. PMID- 15789215 TI - Endoscopic biopsy interpretation difficulties in a congenital diffuse intracranial teratoma. AB - INTRODUCTION: Congenital brain tumours are a rare entity that is nowadays often already recognised during pregnancy by ultrasound (US) and magnetic resonance (MR). Even though the definitive diagnosis is usually achieved by means of histological studies, in some cases the diagnosis may remain uncertain because of the malformative origin of this type of tumour. CASE REPORT: We describe a patient with a diffuse congenital intracranial mass diagnosed by intrauterine US and foetal MR performed to further evaluate the lesion and the associated foetal hydrocephalus. After delivery by caesarean section, an endoscopic biopsy was performed. Several specimens were obtained and resulted in the diagnosis of primitive neuroectodermal tumour (PNET). Despite polychemotherapy, the tumour continued to enlarge and the patient died at 2 months of age. Post-mortem histological examination of the intracranial mass showed a diffuse intracranial teratoma. CONCLUSION: Endoscopic biopsy specimen examination resulted in a diagnosis that was not confirmed by post-mortem findings. The scarce differentiation of teratoma components and their high variability in histomorphology as well as the huge size of the tumoral mass may limit the interpretation of endoscopic biopsy specimens, even when multiple and obtained from different areas. PMID- 15789216 TI - Expression of receptor for advanced glycation end products and HMGB1/amphoterin in colorectal adenomas. AB - Receptor for advanced glycation end products (RAGE) is associated with invasion, metastasis, and poor prognosis in colorectal cancer. We studied the expression of RAGE in colorectal adenomas to elucidate the role of RAGE in cancer development. Expressions of RAGE and high-mobility group box-1 (HMGB1)/amphoterin RAGE ligand were examined in 96 colorectal adenomas using immunohistochemistry and in situ hybridization, respectively. Positivity and expression pattern of RAGE were compared with atypia, histological types, size, and HMGB1/amphoterin expression. Of 96 adenomas, 34 (35%) showed RAGE expression. RAGE positivity was significantly higher in adenomas with severe atypia (18/20, P<0.0001) and large sized adenomas (-15 mm) (18/22, P<0.0001). RAGE expression showed three patterns: cytosolic (n=10), luminal (n=14), and membranous (n=10). Cytosolic pattern was associated with mild atypia and small size (-5 mm). Membranous pattern was associated with severe atypia, villous histological type, and co-expression with overexpressed HMGB1/amphoterin. These results suggest that RAGE expression, especially with membranous pattern, is associated with malignant potential of colorectal adenomas. PMID- 15789217 TI - TASK-3 immunoreactivity shows differential distribution in the human gastrointestinal tract. AB - The presence and distribution of TASK-3 immunopositivity (a channel with potential oncogenic significance) was investigated in the human gastrointestinal system. The immunohistochemical reactions were performed with two commercially available polyclonal antibodies, targeting different epitopes of the channel protein. Experiments conducted on frozen and formalin-fixed samples indicated that the application of a suitable antigen retrieval (AR) technique was essential to produce consistent, strong and reproducible TASK-3-specific immunolabelling of the formalin-fixed tissue. The lack of or inappropriate selection of the AR resulted in false-negative reactions. As for the distribution of the TASK-3 channels, strong immunolabelling was observed in the gastric and large intestinal mucosa, with particularly prominent immunoreactivity of the epithelial cells. In contrast, the smooth-muscle layers demonstrated weak TASK-3 positivity. Intense TASK-3 expression was noted in both the exocrine and endocrine pancreas, but the islets of Langerhans exhibited more powerful reactions. The ductal apparatus of the submandibular gland and lymphocytes situated in pericolonic lymph nodes were also TASK-3 positive. Strong TASK-3 positivity could also be observed in malignant gastrointestinal tumours, with intense nuclear-perinuclear labelling of some of the tumour cells. The present findings suggest that TASK-3 channels may have roles in the gastrointestinal functions, including insular hormone secretion. PMID- 15789218 TI - Natriuretic peptide guanylyl cyclase receptors in the kidney of the Japanese eel, Anguilla japonica. AB - Natriuretic peptides are linked to osmoregulation, cardiovascular and volume regulation in fishes. The peptides bind to two guanylyl-cyclase-linked receptors, natriuretic peptide receptor-A (NPR-A) and NPR-B, to elicit their effects. Atrial natriuretic peptide (ANP) binds principally to NPR-A, whereas C-type natriuretic peptide (CNP) binds to NPR-B. The teleost kidney has an important role in the maintenance of fluid and electrolyte balance; therefore, the location of NPR-A and NPR-B in the kidney could provide insights into the functions of natriuretic peptides. This study used homologous, affinity purified, polyclonal antibodies to NPR-A and NPR-B to determine their location in the kidney of the Japanese eel, Anguilla japonica. Kidneys from freshwater and seawater acclimated animals were fixed overnight in 4% paraformaldehyde before being paraffin-embedded and immunostained. NPR-A immunoreactivity was found on the apical membrane of proximal tubule 1 and the vascular endothelium including the glomerular capillaries. In contrast, NPR-B immunoreactivity was located on the smooth muscle of blood vessels including the glomerular afferent and efferent arterioles, and on smooth muscle tissue surrounding the collecting ducts. No difference in the distribution of NPR-A and NPR-B was observed between freshwater and seawater kidneys. Immunoreactivity was not observed in any tissue in which the antibodies had been preabsorbed. In addition, there was no difference in NPR-A and NPR-B mRNA expression between freshwater-acclimated and seawater-acclimated eels. These results suggest that, although utilizing the same second messenger system, ANP and CNP act on different targets within the kidney and presumably elicit different effects. PMID- 15789219 TI - Ontogeny of estrogen receptor (ER) alpha and its co-localization with pituitary hormones in the pituitary gland of chick embryos. AB - Estrogen is involved in regulating the development and hormone secretion of the anterior pituitary gland following its binding to estrogen receptors (ERs) expressed on pituitary cells. However, the pituitary is comprised of several cell types, and to date, there is no data about the specific cell types expressing ERs in embyonic chick pituitary. We therefore followed, by immunohistochemistry, the ontogeny of the pituitary ER alpha (ERalpha), and the cell types expressing ERalpha throughout chick embryo development. ERalpha immunoreacitivity was restricted to the nuclei of pituitary cells. ERalpha-immunopositive (ERalpha(+)) cells were first detected at embryonic day 6.5 (E6.5), after which ERalpha(+) cells were consistently detected throughout the anterior pituitary gland, although the density of ERalpha(+) cells in the caudal lobe of the pars distalis was higher than that in the cephalic lobe. The proportion of ERalpha(+) cells in the pituitary was about 6% at E8.5; expression increased to 22% by E18.5 of gestation, with no additional change until hatching. Double-labeling of ERalpha and pituitary hormones showed that the dominant cell types expressing ERalpha were gonadotrophs immunopositive for luteinizing hormone (LH); the proportion of ERalpha(+) cells expressing LH increased throughout gestation and reached approximately 57% at hatching. About 2%-6% of thyroid-stimulating-hormone immunopositive and 1%-2% prolactin-immunopositive cells expressed ERalpha at later stages of embryonic development, but no growth-hormone-positive or adrenocorticotropic-hormone-positive cells expressed ERalpha during the embryonic period. Thus, gonadotrophs are the main cell population expressing ERalpha in the anterior pituitary gland of chick embryo, and ERalpha is involved in regulating the development of the pituitary gland and the maturation of the hormone secreting function. PMID- 15789221 TI - The use of discriminant analysis and neural networks to forecast the severity of the Poaceae pollen season in a region with a typical Mediterranean climate. AB - Biological particles in the air such as pollen grains can cause environmental problems in the allergic population. Medical studies report that a prior knowledge of pollen season severity can be useful in the management of pollen related diseases. The aim of this work was to forecast the severity of the Poaceae pollen season by using weather parameters prior to the pollen season. To carry out the study a historical database of 21 years of pollen and meteorological data was used. First, the years were grouped into classes by using cluster analysis. As a result of the grouping, the 21 years were divided into 3 classes according to their potential allergenic load. Pre-season meteorological variables were used, as well as a series of characteristics related to the pollen season. When considering pre-season meteorological variables, winter variables were separated from early spring variables due to the nature of the Mediterranean climate. Second, a neural network model as well as a discriminant linear analysis were built to forecast Poaceae pollen season severity, according to the three classes previously defined. The neural network yielded better results than linear models. In conclusion, neural network models could have a high applicability in the area of prevention, as the allergenic potential of a year can be determined with a high degree of reliability, based on a series of meteorological values accumulated prior to the pollen season. PMID- 15789220 TI - Differential Akt activation in the photoreceptors of normal and rd1 mice. AB - Retinitis pigmentosa is a blinding disease in which unknown mechanisms cause the degeneration of retinal photoreceptors. The retinal degeneration (rd1) mouse is a relevant model for this condition, since it carries a mutation also found in some forms of retinitis pigmentosa. To understand the degenerative process in the rd1 mouse, we must identify the survival and apoptosis-related signaling pathways in its photoreceptors and determine whether signaling differs from that in normal mice. The phosphatidylinositol 3-kinase/Akt kinase pathway promotes survival in several different cell types. The purpose of the present study has been to compare Akt activity in retinal cells of normal and rd1 mice. We have found that, in normal mice, Akt becomes activated in the retina in a developmentally regulated and cell-type-specific fashion, encompassing essentially all retinal cells. In most cell types, once Akt activation has begun, it remains in this state throughout life. An exception is seen in the rod photoreceptors, in which Akt is activated only transiently during their development. The rd1 retina behaves identically in all but one respect, namely that the activation of Akt in rod photoreceptors persists until these cells undergo apoptosis. Thus, Akt may participate in constitutive survival processes in retinal neurons, except in rod photoreceptors in which the role of this pathway may be restricted to the developmental period. However, Akt activation in the rods may be part of a defense mechanism initiated in response to insults, such as the retinal degeneration seen in the rd1 mouse. PMID- 15789222 TI - Persistent shedding of viable SARS-CoV in urine and stool of SARS patients during the convalescent phase. AB - In order to further the present knowledge of the emerging severe acute respiratory syndrome-associated coronavirus (SARS-CoV), 486 different specimens from 54 patients with a clinical diagnosis of SARS were investigated for the presence of viral RNA, and 314 plasma specimens of 73 patients were examined for IgM and IgG antibodies specific against SARS-CoV using an indirect ELISA. Viral RNA was detectable in 28 of the 54 patients tested. Cumulative data showed that 67 of the 73 SARS patients demonstrated seroconversion by week 5 of illness. In contrast, only 1 of 278 healthy subjects enrolled in the study was found to be positive for the IgG antibody. Coexistence of viral RNA in plasma and specific antibodies was simultaneously observed over three consecutive weeks in two critical cases. In three convalescent patients in particular, cultivable SARS-CoV was detected in stool or urine specimens for longer than 4 weeks (29-36 days). These findings suggest that SARS-CoV may remain viable in the excretions of convalescent patients. PMID- 15789223 TI - Expression pattern of cytosolic glutathione peroxidase (cGPx) mRNA during mouse embryogenesis. AB - The selenoprotein cytosolic glutathione peroxidase (cGPx) is ubiquitously distributed in a variety of organs, and its primary function is to protect oxidative damage. To investigate the spatial and temporal expression pattern of cGPx mRNA in embryogenesis, as this has not been studied before, reverse transcription-polymerase chain reaction (RT-PCR) was carried out in a thermal cycler using mouse-specific cGPx primers, and in situ hybridization was performed in whole embryos or embryonic tissues using digoxigenin-labeled mouse cGPx riboprobes. Expression of cGPx mRNA was detected in all the embryos retrieved from embryonic days (EDs) 7.5 to 18.5. On EDs 10.5-12.5, cGPx mRNA was highly expressed in the margin of forelimb and hindlimb buds and dorsally in the cranial neural tube, including the telencephalon, diencephalon, and hindbrain neural tube. On ED 13.5, cGPx mRNA was accumulated especially in vibrissae, forelimb and hindlimb plates, tail, and spinal cord. On EDs 14.5-16.5, cGPx mRNA was found in the developing brain, Rathke's pouch, thymus, lung, and liver. On ED 17.5, the expression of cGPx mRNA was apparent in various tissues such as brain, submandibular gland, vibrissae, heart, lung, liver, stomach, intestine, pancreas, skin, and kidney. In particular, cGPx mRNA was greatly expressed in epithelial linings and metabolically active sites such as whisker follicles, alveolar epithelium of lung, surface epithelium and glandular region of stomach, skin epithelium, and cortex and tubules of kidney. Overall results indicate that cGPx mRNA is expressed in developing embryos, cell-specifically and tissue specifically, suggesting that cGPx may function to protect the embryo against reactive oxygen species and/or hydroperoxides massively produced by the intracellular or extracellular environment. PMID- 15789224 TI - Regression formulae for ab initio and density functional calculated chemical shifts. AB - Linear regression formulae are given for converting 1H and 13C magnetic shielding constants calculated at common ab initio and density functional theory levels of calculation into chemical shifts relative to tetramethylsilane. Accuracies of roughly +/-2.2 ppm (13C) and +/-0.15 ppm (1H) or better are found for the training set for most levels. The highest level calculations do not always give better results than economical standard calculations. PMID- 15789225 TI - Regularity in an environment produces an internal torque pattern for biped balance control. AB - In this paper, we present a control method for achieving biped static balance under unknown periodic external forces whose periods are only known. In order to maintain static balance adaptively in an uncertain environment, it is essential to have information on the ground reaction forces. However, when the biped is exposed to a steady environment that provides an external force periodically, uncertain factors on the regularity with respect to a steady environment are gradually clarified using learning process, and finally a torque pattern for balancing motion is acquired. Consequently, static balance is maintained without feedback from ground reaction forces and achieved in a feedforward manner. PMID- 15789226 TI - A novel phospholipid gemcitabine conjugate is able to bypass three drug resistance mechanisms. AB - We have previously synthesized a phospholipid-gemcitabine conjugate and a phospholipid-cytosine arabinoside conjugate that we tested in different human cancer cell lines. The gemcitabine conjugate was more cytotoxic to the cancer cells tested than the cytosine arabinoside (ara-C) conjugate. The focus here was to elucidate the mechanism of action of the conjugate molecule and its ability to bypass certain drug-resistance mechanisms. In contrast to gemcitabine, the gemcitabine conjugate did not enter the cell via the human equilibrative nucleoside transporter (hENT1). Additionally, the gemcitabine conjugate was not a substrate for the multidrug resistance efflux pump, MDR-1, even though the molecule is more lipophilic. Finally, we showed that deoxycytidine kinase (dCK) was not required for the activation of the gemcitabine conjugate. As expected, cells overexpressing dCK were more sensitive to gemcitabine whereas cells overexpressing dCK were not more sensitive to the gemcitabine conjugate. Taken together, these results suggest that the gemcitabine conjugate may be therapeutically superior to gemcitabine due to the conjugate's ability to bypass three resistance mechanisms that often render gemcitabine ineffective as an anticancer agent. PMID- 15789227 TI - Cytomegalovirus-associated meningoradiculoneuritis after treatment of mantle cell lymphoma with a combination of chemotherapy and rituximab. PMID- 15789228 TI - Microsatellite instability is not an uncommon finding in adult de novo acute myeloid leukemia. AB - To investigate the biologic relevance of microsatellite instability (MSI) in de novo acute myeloid leukemia (AML), 102 consecutive adult patients were analyzed by using a panel of seven microsatellites (BAT25, BAT26, D13S1267, D13S174, D2S123, D5S346 and Mdf15). Frame-shift mutations in the repetitive sequences in the coding region of MSH3, MSH6, BAX, TGFBRII and IGFRII were also investigated by using a fluorescent PCR-based assay. Methylation-specific PCR was used to determine the methylation status of hMLH1 in MSI+ cases. MSH3, MSH6 and MLH1 expression was also analyzed in 68 cases by means of real-time quantitative PCR. MSI was detected in 20 cases: 14 cases had MSI-high (instability of at least two microsatellite markers) and 6 cases corresponded to MSI-low (a single polymorphic marker with instability). Six MSI+ cases showed an associated MLL rearrangement (p=0.002). No single case showed a mutation in the repetitive sequences of the MSH3, MSH6, BAX, TGFBRII and IGFRII genes. Most samples displayed low mRNA levels of the repair genes. hMLH1 promoter was hypermethylated in five MSI+ cases. Overall survival analysis revealed no adverse effect of MSI positivity. These results suggest that MSI may be a common biologic finding in de novo AML. PMID- 15789229 TI - The cutoff value of serum ferritin for the diagnosis of iron deficiency in community-residing older persons. AB - The serum ferritin assay is the best single blood test for the diagnosis of iron deficiency. Previous studies with elderly anemic patients have suggested that ferritin level less than 45 mug/L is indicative of iron deficiency. The subjects of these studies were hospitalized patients with anemia, however. We thus conducted a prospective study to determine the normal minimum level of serum ferritin of community-dwelling older adults by assessing the ratio of serum transferrin receptor to the log ferritin level (sTfR-F index). We conducted the anemia survey between October and November 2002. A total of 1,254 apparently healthy older adults, aged between 60 and 95 years, from three urban community dwellings participated in the survey. Among these individuals, 156 subjects who were anemic or whose serum ferritin level was less than 100 microg/L were selected. The soluble transferrin receptor assay was performed and the sTfR-F index was calculated. The receiver operating characteristic curve analysis was performed. Based on the data, serum ferritin level of 22 microg/L was selected as the cutoff value for the diagnosis of iron deficiency in community-dwelling older adults. Applying the serum ferritin cutoff of 22 microg/L and the sTfR-F index cutoff of 1.5, the sensitivity of the assay was 89.5% (34 of 38) and the specificity was 89.0% (105 of 118). In conclusion, for the diagnosis of iron deficiency of community-residing older adults, we suggest the serum ferritin cutoff value of 22 microg/L obtained by use of the sTfR-F index. The value is lower than the previous value established for hospitalized and anemic older adults. PMID- 15789230 TI - Anthropometric measurements and growth as predictors of low-back pain: a cohort study of children followed up from the age of 11 to 22 years. AB - Body height is an alleged risk factor for low-back pain (LBP) in adulthood, but its importance is obscure during childhood and adolescence. We studied growth for its association with the incidence of LBP in a population study of 430 children who were examined five times: at the age 11,12,13,14 and 22 years. Body height and weight and the degrees of trunk asymmetry, thoracic kyphosis and lumbar lordosis were measured at every examination. The history of LBP was obtained by a structured questionnaire at the ages of 14 and 22 years. The incidence of LBP was defined as pain, which occurred on eight or more days during the past year among those 338 children who had been free from LBP until 14 years of age. Growth of body height between 11 years and 14 years of age predicted the incidence of LBP. Adjusted for sex, the odds ratio (with 95% confidence interval) per an increment of one SD (4.3 cm) was 1.32 (1.06--1.65), the P value for trend being 0.03. Growth after 14 years of age was inversely related to the incidence of LBP, but the association did not reach statistical significance (P for trend=0.06). Other anthropometric measurements or their changes were not found to predict LBP. Our results are not compatible with the old myth that spinal growth actually contributes to LBP. But abundant growth in early adolescence may be a risk factor for subsequent LBP. PMID- 15789231 TI - Correlative radiological, self-assessment and clinical analysis of evolution in instrumented dorsal and lateral fusion for degenerative lumbar spine disease. Autograft versus coralline hydroxyapatite. AB - This prospective longitudinal randomized clinical and radiological study compared the evolution of instrumented posterolateral lumbar and lumbosacral fusion using either coralline hydroxyapatite (CH), or iliac bone graft (IBG) or both in three comparable groups, A, B and C, which included 19, 18 and 20 patients, respectively, who suffered from symptomatic degenerative lumbar spinal stenosis and underwent decompression and fusion. The patients were divided randomly according to the graft used and the side that it was applied. The spines of group A received autologous IBG bilaterally; group B, IBG on the left side and hydroxyapatite mixed with local bone and bone marrow on the right side; group C, hydroxyapatite mixed with local bone and bone marrow bilaterally. The age of the patients in the groups A, B and C was 61+/-11 years, 64+/-8 years and 58+/-8 years, respectively. The SF-36, Oswestry Disability Index (ODI), and Roland Morris (R-M) surveys were used for subjective evaluation of the result of the surgery and the Visual Analogue Scale (VAS) for pain severity. Plain roentgenograms including anteroposterior, lateral and oblique views, and lateral plus frontal bending views of the instrumented spine and CT scan were used to evaluate the evolution of the posterolateral fusion in all groups and sides. Two independent senior orthopaedic radiologists were asked to evaluate first the evolution of the dorsolateral bony fusion 3-48 months postoperatively with the Christiansen's radiologic method, and secondly the hydroxyapatite resorption course in the spines of groups B and C. The diagnosis of solid spinal fusion was definitively confirmed with the addition of the bending views, CT scans and self assessment scores. The intraobserver and interobserver agreement (r) for radiological fusion was 0.71 and 0.69, respectively, and 0.83 and 0.76 for evaluation of CH resorption. T(12)-S(1) lordosis and segmental angulation did not change postoperatively. There was no radiological evidence for non-union on the plain roentgenograms and CT scans. Radiological fusion was achieved 1 year postoperatively and was observed in all groups and vertebral segments. Six months postoperatively there was an obvious resorption of hydroxyapatite granules at the intertransverse intersegmental spaces in the right side of the spines of group B and both sides of group C. The resorption of hydroxyapatite was completed 1 year postoperatively. Bone bridging started in the third month postoperatively in all instrumented spines and all levels posteriorly as well as between the transverse processes in the spines of the group A and on the left side of the spines of group B where IBG was applied. SF-36, ODI, and R-M score improved postoperatively in a similar way in all groups. There was one pedicle screw breakage at the lowermost instrumented level in group A and two in group C without radiologically visible pseudarthrosis, which were considered as having non-union. Operative time and blood loss were less in the patients of group C, while donor site complaints were observed in the patients of the groups A and B only. This study showed that autologous IBG remains the "gold standard" for achieving solid posterior instrumented lumbar fusion, to which each new graft should be compared. The incorporation of coralline hydroxyapatite mixed with local bone and bone marrow needs adequate bleeding bone surface. Subsequently, hydroxyapatite was proven in this series to not be appropriate for intertransverse posterolateral fusion, because the host bone in this area is little. However, the use of hydroxyapatite over the decorticated laminae that represents a wide host area was followed by solid dorsal fusion within the expected time. PMID- 15789232 TI - Lessons learned from cervical pseudoarthrosis in ankylosing spondylitis. AB - This case report illustrates three learning points about cervical fractures in ankylosing spondylitis, and it highlights the need to manage these patients with the neck initially stabilised in flexion. We describe a case of cervical pseudoarthrosis that is a rare occurrence after fracture of the cervical spine with ankylosing spondylitis. This went undetected until the development of myelopathic symptoms many months later. The neck was initially stabilised in flexion using tongs, and then slowly extended before anterior and posterior fixation was performed. The myelopathic symptoms resolved, and the patient had a good result at 18 months. We conclude that any increased movement of the spine after trauma in ankylosing spondylitis must be considered suspect and fully investigated. PMID- 15789233 TI - Role of parenteral iron in the management of anaemia in the elderly patient undergoing displaced subcapital hip fracture repair: preliminary data. AB - INTRODUCTION: An important percentage of patients affected by hip fracture require the administration of allogenic blood transfusion (ABT) to avoid the risks of perioperative acute anaemia. However, concerns about ABT risks have led to the search for alternatives, especially in elective orthopaedic surgery. MATERIALS AND METHODS: We have prospectively investigated the effect of preoperative intravenous 200-300 mg (group 2; n=20) iron sucrose on ABT requirements and postoperative morbid-mortality in patients undergoing surgery for displaced subcapital hip fracture (DSHF) repair. A previous series of 57 DSHF patients served as the control group (group 1). All patients were older than 65 years, were operated on the 3rd day after admission to the hospital, by the same medical team, and using the same implant. Age, gender, ASA classification, surgical procedure, perioperative haemoglobin, requirements for ABT, postoperative infection, length of hospital stay (LOS) and 30-day mortality rate were examined. RESULTS: No adverse reactions to the iron administration were observed. The iron group had a lower transfusion rate (15% vs 36.8%), lower transfusion index (0.26 vs 0.77 units per patient), lower 30d mortality rate (0 vs 19.3%), shorter LOS (11.9 vs 14.1 days), as well as a trend to a lower postoperative infection rate (15% vs 33%). CONCLUSION: Preoperative parenteral iron administration could be a safe and effective way to reduce the ABT requirements in DSHF patients. This reduction in the ABT requirements is accompanied by a reduction in the morbid-mortality rate and LOS. A large, randomised, controlled trial to confirm these results is warranted. PMID- 15789234 TI - Influence of pH on copper and zinc sensitivity of ericoid mycobionts in vitro. AB - The effect of pH on growth, metal uptake and toxicity in four isolates of ericoid mycobionts (two Hymenoscyphus ericae from unpolluted heathland sites and two H. ericae-type mycobionts from metal-contaminated mine spoil) was assessed in vitro. These isolates were incubated in liquid medium (10% Rorison's medium, glucose at 10 g l(-1)) containing either 0.25 mM Cu or 2.0 mM Zn and adjusted to pH 2, 3, 4, 5 or 6. After 30 days incubation, dry mass and mycelial metal content were determined and growth was expressed as tolerance index, i.e. dry mass in the presence of metal as a percentage of dry mass in the absence of metal. Initial medium pH had a significant effect on both tolerance index and metal accumulation. Tolerance indices were highest at pH 2, with several isolates showing a stimulation of growth (i.e. tolerance index >100%) at this pH. Tolerance index decreased at higher initial pH values and growth of two mycobionts was completely inhibited (tolerance index=0) in the Cu-supplemented media at pH 6. Reduction in tolerance index coincided with an increase in mycelial accumulation of Cu and Zn. Practical and environmental implications of these results are discussed. PMID- 15789235 TI - Serum-free generation and quantification of functionally active Leukemia-derived DC is possible from malignant blasts in acute myeloid leukemia and myelodysplastic syndromes. AB - Functional dendritic cells (DC) are professional antigen presenting cells (APC) and can be generated in vitro from leukemic cells from acute myeloid leukemia AML patients, giving rise to APC of leukemic origin presenting leukemic antigens (DC(leu)). We have already shown that DC can be successfully generated from AML and myeloplastic syndromes (MDS) cells in serum-free 'standard' medium (X-vivo + GM-CSF + IL-4 +TNFalpha + FL) in 10-14 days. In this study, we present that DC counts generated from mononuclear cells (MNC) varied between 20% (from 55 MDS samples), 34% (from 100 AML samples) and 25% (from 38 healthy MNC samples) medium. Between 53% and 58% of DC are mature CD83+ DC. DC harvests were highest in monocytoid FAB types (AML-M4/M5, MDS-CMML) and independent from cytogenetic risk groups, demonstrating that DC-based strategies can be applied for patients with all cytogenetic risk groups. Proof of the clonal derivation of DC generated was obtained in five AML and four MDS cases with a combined FISH/immunophenotype analysis (FISH-IPA): The clonal numerical chromosome aberrations of the diseases were regularly codetectable with DC markers; however, not with all clonal cells being convertible to leukemia-derived DC(leu) (on average, 53% of blasts in AML or MDS). To the contrary, not all DC generated carried the clonal aberration (on average, 51% of DC). In 41 AML and 13 MDS cases with a suitable antigen expression, we could confirm FISH-IPA data by Flow cytometry: although DC(leu) are regularly detectable, on average only 57% of blasts in AML and 64% of blasts in MDS were converted to DC(leu). After coculture with DC in mixed lymphocyte reactions (MLR), autologous T cells from AML and MDS patients proliferate and upregulate costimulatory receptors. The specific lysis of leukemic cells by autologous T cells could be demonstrated in three cases with AML in a Fluorolysis assay. In six cases with only few DC(leu) or few vital T cells available after the DC/MLR procedure, no lysis of allogeneic or autologous leukemic cells was seen, pointing to the crucial role of both partners in the lysis process. We conclude: (1) the generation of DC is regularly possible in AML and also in MDS under serum-free conditions. (2) Clonal/leukemia-derived DC(leu) can be regularly generated from MDS and AML-MNC; however, not with all blasts being converted to DC(leu) and not all DC generated carrying leukemic markers. We recommend to select DC(leu) for vaccinations or ex vivo T-cell activations to avoid contaminations with non-converted blasts and non-leukemia-derived DC and to improve the harvest of specific, anti-leukemic T cells. DC and DC-primed T cells could provide a practical strategy for the immunotherapy of AML and MDS. PMID- 15789236 TI - Tubularized incised plate urethroplasty for abortive hypospadiac urethral duplication. AB - Urethral duplication is among the rarest congenital anomalies and is characterized by two urethral canals. The urethral canals are most commonly located in the sagittal plane. We present a case of abortive hypospadiac urethral duplication and its surgical management with tubularized incised plate urethroplasty. PMID- 15789237 TI - Mesenteric inflammatory pseudotumor as a cause of abdominal pain in a teenager: presentation and literature review. AB - Inflammatory pseudotumor (IP) is an unusual cause of chronic abdominal pain in children. The management of these lesions is complicated by controversies surrounding their appropriate classification and the numerous alternate names with which they are described. Successful treatment requires careful radiologic and pathologic evaluation to distinguish IPs from other lesions, along with complete surgical resection. We present the case of a 15-year-old boy with IP and review the literature in an attempt to simplify the description of these tumors. PMID- 15789238 TI - Giant pedunculated lipofibroma of the elbow. AB - Lipofibromas are rare hamartomatous lesions almost exclusively involving the medial nerve and have been reported to date only in the adult population. The case of a giant pedunculated lipofibroma involving the radial nerve in the elbow of a 1-year-old child is presented. PMID- 15789239 TI - Bronchogenic cyst of the lung mimicking a pulmonary hydatid cyst in a child. AB - Lung bronchogenic cyst may be infected and present with symptomatology and chest radiological findings compatible with an infected lung cyst, including pulmonary hydatid cyst. We describe a case of a 5-year-old girl with a symptomatic solitary mass in the right lower lobe that was initially treated as a hydatid cyst. Eventually, a lobectomy was performed, and a final diagnosis done on pathology showed an infected congenital bronchogenic cyst. PMID- 15789240 TI - Extrapleural thoracoscopic repair of esophageal atresia with tracheoesophageal fistula. AB - Traditionally, open repair of esophageal atresia (EA) with tracheoesophageal fistula (TEF) required thoracotomy. Innovations in minimal access surgery have created a thoracoscopic technique resulting in violation of the pleural space. Most pediatric surgeons favor an extrapleural approach for open repair. We present a novel minimal access, extrapleural technique for repairing EA with TEF. A 2-day-old infant with EA and distal TEF underwent thoracoscopic extrapleural repair that utilized three ports. Initial creation of the extrapleural space was achieved through one of the port sites and was completed thoracoscopically. A thoracoscopic repair of EA with distal TEF was achieved within the extrapleural space. A small tear in the pleura was inadvertently created during the dissection. The child began feeding normally. At 1 year of age, the patient had dysphagia requiring a single esophageal dilatation. This is the first known report of an extrapleural thoracoscopic repair of EA with TEF. Although thoracoscopic repairs of EA/TEF have been previously reported, these were all done transpleurally. Many pediatric surgeons favor the extrapleural approach for two reasons: (1) containment of a potential leak within the extrapleural space, avoiding an empyema, and (2) easier transpleural access for future thoracic procedures. PMID- 15789241 TI - Evolution of ochratoxin A content from must to wine in Port Wine microvinification. AB - To study the evolution of ochratoxin A (OTA) content from must to wine during the making of Port Wine, grapes from the five most common varieties of Port Wine were harvested and combined in equal percentages in order to perform microvinifications. Three sets of assays were studied: a blank (A), where the most common Port Wine-making process was used; in the second (B), a solution of OTA was added to the initial must; in the third (C), the grapes were aspersed with an inoculating solution of OTA-producing fungi. Samples were collected, in duplicate, on four different occasions throughout the process. The influence of the addition of SO2 to the must was also assessed in each set. The quantification of OTA was based on the standard reference method for wines (European Standard prEN 14133), which includes clean-up via immunoaffinity columns and HPLC with fluorescence detection. The limits of detection were 0.076 microg/l for wine and 0.114 microg/l for must. The method was validated by assessing the precision, accuracy and by obtaining an estimate of the global uncertainty. Overall, the levels of OTA observed during the vinifications dropped by up to 92%, and no grapes used in this work were contaminated naturally. PMID- 15789244 TI - Detector for particulate polycyclic aromatic hydrocarbons in water. AB - It is estimated that most polycyclic aromatic hydrocarbons (PAHs) in environmental water are not dissolved but rather in particulate form. Nevertheless, the currently available optical detectors are not suited for proper sampling of solid PAHs. A new setup for direct sampling and quantification of suspended particulate PAHs in water is suggested. It is based on a polymeric film that has the capability of dissolving PAH particulates, coupled to a traditional laser-induced fluorescence probe. Kinetics and performance of two sampling modes have been studied: bulk sampling, by immersing the probe into the water, and surface sampling, by laying the film on the water surface. The latter method has proved to be more sensitive; however, it is diffusion-limited. Linear calibration plots have provided quantification over a wide concentration range with detection limits in the ppb range (these could be improved by using a modified probe). The effects due to other particulates in water have been studied and only little interferences have been observed. The possibility of analysis of PAH mixtures has been addressed and it has been concluded that multivariate analysis is needed. PMID- 15789245 TI - Factors affecting signal intensity in headspace mass spectrometry for the determination of hydrocarbon pollution in beach sands. AB - One of the main limitations to the use of direct coupling of headspace mass spectrometry (HS-MS) for the quantitative determination of analytes in a sample is related to factors affecting the signal intensity. The importance of strategies aimed at compensating this problem is considerable in the case of classification, and is indeed critical as regards the problems involved in quantification. This paper reports the effects of the different factors affecting HS-MS signal intensity in the quantification of the pollution of beach sands by hydrocarbons--the matrix effect, signal instability over time and nature of the different pollutants present in the polluted sands--and proposes possible solutions. Signal instability was solved by using a multiplicative calibration transfer algorithm. A three-factor Box-Behnken experimental design was used to study the matrix effect, mainly as regards the moisture of the samples, and the results are discussed. PMID- 15789246 TI - Frequent analytical/experimental problems in lipase-mediated synthesis in solvent free systems and how to avoid them. AB - Compared with chemical catalysis, enzymatic catalysis is a relatively new topic. Experimental work involving lipases deserves careful attention and accurate procedures still need to be implemented. A rapid but careful survey of published data immediately demonstrates that experiments performed under similar conditions with similar reagents have led to very different results. The aim of this work is to point out the importance of accurate and systematic procedures in order to ensure the reproducibility of experimental data. We strongly believe that different results found by different labs are due to problems detected in the procedures used. Quantification of the immobilisation efficiency of lipase on several supports through UV/visible methods and sampling methods used to obtain correct enzymatic activity values are specifically analysed. After a brief review which demonstrates the big discrepancies found in the literature, original data from Candida rugosa lipase adsorption on polypropylene powder and its use in the solvent-free synthesis of ethyl oleate are introduced in order to exemplify the difficulties found in these kinds of systems. Several procedures described in the literature are assayed and the accuracy of the results obtained is carefully analysed. The aim of the whole analysis performed is that it would be useful for any powdered solid to be used as a support for a lipase in a solvent-free system for any synthesis reaction, especially for those involving a volatile reagent. Throughout this contribution, special emphasis is placed on how catalytic reaction results using enzymes (free and immobilised) are reported so as to allow comparison between published data, something which is usually difficult since very different units are used and often complementary data are not included. PMID- 15789247 TI - Restricted expression of a median Hox gene in the central nervous system of chaetognaths. AB - Hox genes encode a set of evolutionarily conserved transcription factors that regulate anterior-posterior patterning. Here we report the first developmental expression of a Hox gene from Chaetognatha. These metazoans have been shown recently to be part of the protostome group of bilaterians. We describe the analysis of the SceMed4 gene (a Spadella cephaloptera Median Hox gene) including its expression from late stages of egg development to 7 days after hatching. In all of these stages, SceMed4 is expressed in two lateral stripes in a restricted region of the developing ventral ganglion. PMID- 15789248 TI - Phylogenetic analysis and developmental expression of thymosin-beta4 gene in amphioxus. AB - Thymosin-beta4 is a highly conserved actin-binding protein that plays an important role in multiple early developmental events and functions in keeping the adult life in vertebrates. Here a cDNA for a thymosin-beta4 gene was isolated from the amphioxus, Branchiostoma belcheri. A molecular phylogenetic tree constructed from the deduced amino acid sequence of the isolated cDNA indicates that this gene belongs to the thymosin-beta4 subfamily, but it is split at the base of the vertebrate gene clade in evolution. In situ hybridization reveals that the expression is detected in the locations homologous to orthologous genes expressing regions of vertebrate embryos and adults, such as the neural plate, neural tube, paraxial mesoderm, differentiating somites, pharynx and gut, midgut diverticulus, blood vessels and body spaces. These results are interpreted to mean that thymosin-beta4 genes might play a conserved role in the patterning of chordate embryos and functions in adults. PMID- 15789249 TI - Low-dose CT colonography in children: initial experience, technical feasibility, and utility. AB - BACKGROUND: CT colonography (CTC) is utilized as a diagnostic tool in the detection of colon polyps and early colorectal cancer in adults. Large studies in the literature, although focused on adult populations, have shown CTC to be a safe, accurate, non-invasive technique. OBJECTIVE: We evaluated the technical feasibility of CTC in children using a low-dose technique. MATERIALS AND METHODS: From November 2001 to April 2004 we evaluated eight patients (3-17 years) with non-contrast CTC. Seven of the patients had CTC, followed by standard colonoscopy (SC) the same day; in one patient, CTC followed a failed SC. CTC results were compared to results of SC. The estimated effective dose from each CTC was calculated and compared to that of standard barium enema. RESULTS: CTC results were consistent with those of SC. Sensitivity for polyps 5-10 mm was 100%, and sensitivity for polyps 10 mm and larger was 66.7%. The estimated mean effective dose was 2.17 mSv for CTC, compared to the 5-6 mSv for a standard air-contrast barium enema in a small child. CONCLUSION: Our initial experience shows CTC in children is well-tolerated, safe, and useful. The procedure can be performed successfully with a low radiation dose, and preliminary results compare well with SC. PMID- 15789250 TI - [Traumatic anterior shoulder dislocation]. AB - The purpose of this paper is to outline the present state of treatment for traumatic anterior shoulder dislocation. Pathoanatomic changes as well as intrinsic and extrinsic prognostic factors are discussed. Based on these, treatment options for first time traumatic dislocations and chronic posttraumatic instability are recommended. PMID- 15789251 TI - The effects of memantine on lipid peroxidation following closed-head trauma in rats. AB - Memantine is an uncompetitive N-methyl-D: -aspartate (NMDA) receptor antagonist. Unlike other NMDA antagonists, it has been used clinically for years for the treatment of Parkinson's disease, spasticity, and dementia without serious side effects. We aimed to investigate the therapeutic efficacy of memantine on a closed head trauma model. A total of 132 adult male Sprague-Dawley rats were randomly divided into four groups: sham-operated, control (closed head trauma), sham-vehicle (closed head trauma + saline), treatment (closed head trauma + memantine, 10 mg/kg, i.p.). A cranial impact was delivered to the skull, just in front of the coronal suture, over the left hemisphere, from the height of 7 cm. Saline or memantine were applied 15 min after trauma. Rats were euthanased 0.5, 1, 2, 6, 24, 48 h after trauma. Brain tissue samples were taken 5 mm away from the left frontal pole and also from the corresponding point of the contralateral hemispheres. Malondialdehyde activity (MDA) was considered to reflect the degree of lipid peroxidation. The MDA levels continued to increase for the first 2 h after the injury, then started to decrease gradually. Memantine treatment significantly reduced lipid peroxidation levels in the treatment group compared with other groups (P<0.01). The findings of the present study indicate that memantine provides beneficial effects after closed head trauma in rats. PMID- 15789252 TI - Acute brainstem dissection of syringomyelia associated with cervical intramedullary neurinoma. AB - Intramedullary tumors and syringomyelia typically present with slowly progressing deficits. More rarely, they are characterized by acute presentation or worsening, at times mimicking other more common etiologies. The acute onset of syringomyelia is most likely attributable to an acute increase in cerebrospinal fluid and epidural venous pressure that results in impulsive fluid movement and, ultimately, in the rupture of the syrinx and dissection into the spinal cord or brainstem. Reported here is a case of acute presentation of a small cervical intramedullary neurinoma due to the upward dissection of its associated syrinx. Critical questions are: (1) how can a small tumor produce a large syrinx? and (2) in the absence of craniospinal interferences, which mechanism underlies the acute expansion of the cavity, resulting in a rapid onset? The authors examined the pathophysiology of syrinx formation and enlargement in intramedullary tumors and reviewed the literature, emphasizing the relationship between spinal cord movements and intramedullary pressure. On the basis of current pathogenetic concepts, the authors concluded that tumor-related syringomyelia might be caused by an association of mechanisms, both from within (obstruction of perivascular spaces; increase in extracellular fluid viscosity due to the tumor itself; intramedullary pressure gradients among different cord levels and between the cord and the subarachnoid space) and from without (the cerebrospinal fluid entering the tissue). All these factors may be amplified, as in the reported case, by a tumor located dorsally at the cervical level. Abnormal postures of the spine, such as a prolonged and excessive flexed neck position, may ultimately contribute to the acute dissection of the syrinx. PMID- 15789253 TI - Effect of maturational status and training on upper limb pulse wave velocity. AB - Cardiovascular adaptations to training may include changes in arterial distensibility, although studies involving sick and healthy individuals have yielded contradictory results. Moreover, despite the fact that endurance training frequently begins in childhood, its influence on arterial distensibility is less well characterised in children. This study was therefore to compare the upper limb pulse wave velocity (PWVUL) in highly trained prepubertal swimmers and age matched moderately active counterparts, with that in swimmers and sedentary adults. The aim was to investigate the effects of maturation and training status on PWVUL in healthy individuals. PWVUL was evaluated by a dedicated continuous wave Doppler and photoplethysmography system. We calculated the pressure corrected index of distensibility (Cp) which provides a measure of intrinsic structural compliance of the artery wall. PWVUL was significantly higher in highly trained child swimmers compared to moderately active ones (6.17+/-0.49 versus 5.20+/-0.38 ms(-1), P <0.05) even when mean arterial pressure was fixed as a covariate. Cp was significantly lower in child swimmers than in moderately active ones (0.15+/-0.03 versus 0.18+/-0.04, P <0.05 respectively) whereas Cp was not significantly different in either adult group. The PWVUL and Cp values of child swimmers were similar to those of adults. CONCLUSION: Our results confirmed an increase in pulse wave velocity with maturation implying that compliance decreases with age. An unexpected result was that swimming training performed in children over a long period led to an increase in upper limb pulse wave velocity. This change, concerning the upper limbs which are strongly involved in swimming, may be related to enhanced smooth muscle content of the arterial wall, possibly due to intermittent elevations in arterial blood pressure during repetitive swimming exercise sessions. Whether adaptations to intensive swimming training are accompanied by alterations of the cardiovascular system in prepubertal children is an important question to be dealt with in further studies. PMID- 15789254 TI - Retinol binding protein status in relation to ocular surface changes in patients with cystic fibrosis treated with daily vitamin A supplements. AB - Cystic fibrosis (CF) is an autosomal recessive disease characterised by increased viscosity of mucus secretions and high chloride concentration in exocrine secretions. Clinically, the patients suffer from chronic pulmonary changes, chronic pancreatic deficiency, and an obstruction of the gastrointestinal tract. The disease affects all secretory epithelia including the eye. The influence of nutritional status on long-term survival and quality of life of CF patients is well documented. Steatorrhea, a consequence of decreased fat digestion and absorption may be associated with vitamin deficiences, including vitamin A. The aim of this study was to document plasma retinol binding protein (RBP) status, a specific plasma transport protein for vitamin A, and ocular surface changes in children and adolescents with CF. The patients were recruited at the 3rd Department of Paediatric Diseases, Medical University of Bialystok, Poland. All patients were regularly seen by a CF specialist dietitian. A group of 15 patients had the following investigations: plasma RBP, visual acuity, physical examination, tear film break-up time, fluorescein staining and Schirmer tear test. A group of 15 age- and sex-matched controls without CF or ocular pathology were also recruited. Plasma RBP concentrations were significantly lower in patients with CF than in the control group. CF patients showed a statistically significant increase in the incidence of clinical blepharitis. Five of the CF patients had clinical evidence of dry eyes. CONCLUSION: Low plasma retinol binding protein levels frequently occur in clinically stable and retinol supplemented cystic fibrosis patients, of whom five had dry eyes. We recommend monitoring of plasma retinol binding protein levels and evaluation of ocular surface changes, especially those with dry eye symptoms in all cystic fibrosis patients. PMID- 15789256 TI - Laparoscopic colectomy for colon adenocarcinoma: an 11-year retrospective review with 5-year survival rates. AB - BACKGROUND: Laparoscopic colectomy for the management of colon cancer remains a controversial therapeutic option, especially when the outcomes are compared with the historically accepted survival data and recurrence rates after open surgery. The purpose of this study was to evaluate the 5-year overall and disease-free survival rates after laparoscopic colon resection for invasive colon adenocarcinoma. METHODS: A total of 129 patients underwent consecutive laparoscopic colectomies for colon adenocarcinoma (between April 1992 and 2004 January) by a single surgeon at a single institution. Records were analyzed retrospectively and follow-up data was obtained. The Student t-test, Cox regression analysis, and Kaplan-Meier survival data were used for statistical analysis. RESULTS: After patients with noninvasive disease on final pathology were excluded, the study population comprised 88 patients who underwent laparoscopic colectomies for invasive colon cancer with > 2 years of follow-up. Of these cases, 81 (93%) were amenable for complete follow-up at 11years (41 women and 40 men; mean age, 76 years). Mean follow-up was 61 months. There was one perioperative death (1.2%), and the overall postoperative morbidity rate was 13.6%. The average number of lymph nodes harvested was 10.1 (+/-6). There were no port site recurrences. The Kaplan-Meier survival data were as follows for 5-year overall survival and 5-year disease-free survival, respectively stage I (n = 34) 89% and 89%; stage II (n = 22), 65% and 59%; stage III (n = 19), 72% and 67%; stages I-III combined, (n = 75), 77% and 73%. CONCLUSIONS: For this specific cohort of patients undergoing curative laparoscopic colectomies for invasive colon adenocarcinoma, the mean follow-up was > 5 years. Overall survival and disease-free survival for stage I, II, and III colon cancer as well as for stages I-III combined are favorable and comparable to historically acceptable open colectomy survival rates. Overall survival and disease-free survival after laparoscopic colectomy for invasive colon cancer is no worse, and perhaps better than, the previously reported rates for the same procedure done by an open technique. PMID- 15789257 TI - Full-face undermining using progressive dilators. AB - The application of a new and simple method of facial cutaneous detachment using specially designed bifaceted rigid steel wands called Dilson Luz Vascular Dilation Wands is described. These wands considerably reduce the bleeding during face-lift surgery, reducing both postoperative hematomas and the risk of facial nerve injury. This technique involves stretching the blood vessels to the point of rupture by inserting progressively larger wands. At the point of rupture, a spontaneous coagulation within the vascular extremities occurs. This is attributable to a significant blood platelet migration induced by the stimulation from the rupture of the vascular intima. The wands, varying in dimension from 1.5 to 20 mm, and are used for cutaneous facial detachment. This method has been found to result in minimal bleeding across the detached surface as well as an excellent perfusion of the overlying skin. With the use of these Dilson Luz Vascular Dilation Wands in combination with this new technique, the authors have obtained improved cutaneous detachment; reduced postoperative swelling, edema, and ecchymosis; prophylaxis of facial nerve damage; reduced procedural time; and above all, greater reduction in immediate and late postoperative hematoma formation. PMID- 15789255 TI - Comparison of endoscopic techniques vs Shouldice and other open nonmesh techniques for inguinal hernia repair: a meta-analysis of randomized controlled trials. AB - BACKGROUND: We performed a scientific evaluation of the efficacy of different surgical techniques for inginual hernia repair and supported our findings by conducting a systematic review of randomized studies comparing endoscopic with open nonmesh suture techniques. METHODS: After an extensive search of the literature, a total of 27 studies (41 publications) with evidence level lb were identified. These studies randomly compared endoscopic with open nonmesh suturing techniques. The quality of data sufficed to enable a quantitative meta-analysis of various parameters using the original software of the Cochrane Collaboration. Due to its superiority in comparison to other open nonmesh suturing techniques, the Shouldice repair technique was analyzed separately. RESULTS: The systematic comparison of endoscopic techniques with the Shouldice repair showed that these techniques had significant advantages in terms of the following parameters: total morbidity, hematoma, nerve injury, and pain-associated parameters such as time to return to work, and chronic groin pain. The Shouldice operation has the advantages of a shorter operating time and a lower incidence of wound seroma. There was no difference regarding the incidence of major complications, wound infection, testicular atrophy, or hernia recurrence. Open non-Shouldice suturing techniques are associated with higher recurrence rates and more wound infections than endoscopic operations. CONCLUSION: In comparison to open nonmesh suture repair techniques, endoscopic repair techniques have significant advantages in terms of pain-associated parameters. For the revaluation of long-term complications such as hernia recurrence and chronic groin pain, further clinical examination of the existing study collectives is needed. PMID- 15789258 TI - Primary stenting of intracranial atherosclerotic stenoses. AB - PURPOSE: To determine the feasibility and safety of stenting intracranial atherosclerotic stenoses. METHODS: In 12 patients the results of primary intracranial stenting were evaluated retrospectively. Patient ages ranged from 49 to 79 years (mean 64 years). Six patients presented with stenoses in the anterior circulation, and six had stenosis in the posterior circulation. One patient presented with extra- and intracranial tandem stenosis of the left internal carotid artery. Three patients presented with acute basilar thrombosis, caused by high-grade basilar stenoses. RESULTS: Intracranial stenoses were successfully stented in 11 of 12 patients. In one patient the stent could not be advanced over the carotid siphon to reach the stenosis of the ophthalmic internal carotid artery. Follow-up digital subtraction angiographic studies were obtained in two patients who had presented with new neurologic signs or symptoms. In both cases the angiogram did not show any relevant stenotic endothelial hyperplasia. In one patient, after local thrombolysis the stenosis turned out to be so narrow that balloon angioplasty had to be performed before stent deployment. All three patients treated for stenosis-related basilar thrombosis died due to brainstem infarction that had ensued before the intervention. CONCLUSIONS: Prophylactic primary stenting of intracranial stenoses of the anterior or posterior cerebral circulation can be performed with a low complication rate; technical problems such as stent flexibility must still be solved. Local thrombolysis followed by stenting in stenosis-related thrombotic occlusion is technically possible. PMID- 15789259 TI - Ex vivo experiment of saline-enhanced hepatic bipolar radiofrequency ablation with a perfused needle electrode: comparison with conventional monopolar and simultaneous monopolar modes. AB - The purpose of this study was to validate the saline-enhanced bipolar radiofrequency ablation (RFA) technique using a perfused electrode to increase RF created coagulation necrosis, to compare that technique with monopolar RFAs and to find appropriate concentrations and volumes of perfused NaCl solution for the bipolar RFA. A total of 90 ablations were performed in explanted bovine livers. In the initial experiments to determine appropriate conditions for bipolar RFA, we created five thermal ablation zones in each condition, with instillations of varied concentrations (0.9-36%) or injection rates (30 mL/hr-120 mL/hr) of NaCl solution. After placement of one or two 16-gauge open-perfused electrodes into bovine livers, the NaCl solution was instilled into the tissue through the electrode. In the second part of the study, 10 ablation zones were created using one or two perfused electrodes for each of five groups under different conditions: a conventional monopolar mode with 0.9% NaCl solution (group A) or with 6% NaCl solution (group B), a simultaneous monopolar mode with 6% NaCl solution (group C) and a bipolar mode with 6% NaCl solution (groups D and E). RF was applied to each electrode for 20 min in groups A, B, C, and E, or for 10 min in group D. During RFA, we measured the tissue temperature 15 mm from the electrode. The temperature changes during the RFA and the dimensions of the ablation zones were compared among the groups. Bipolar RFA created larger short axis diameters of coagulation necrosis with 6% NaCl solution (35.8 +/- 15 mm) than with 0.9% NaCl solution (17 +/- 9.7 mm) (P < 0.05). However, concentrations of NaCl solution above 6% did not further increase the extent of coagulation necrosis. In addition, bipolar RFA with 6% NaCl solution instillation at a rate of 1.0 mL/min (37.9 +/- 5.4 mm) or 2.0 mL/min (35.6 +/- 9.3 mm) produced larger diameters at the mid-point between the electrodes of the ablated lesion than did 0.5 mL/min (25.8 +/- 9.3 mm) (P < 0.05). The bipolar mode showed a more rapid increase in temperature at the mid-point between the two probes, up to 60 degrees C, than did the monopolar modes (P < 0.05). In addition, the bipolar RFA (group E) treated for 20 min showed a larger value of the short-axis diameter than did the conventional or simultaneous monopolar modes (P < 0.05), and bipolar RFA (group D) treated for 10 min, showed similar results with conventional monopolar modes treated for 20 min (P > 0.05): 31.0 +/- 5.4 mm (group A); 28.8 +/- 3.8 mm (group B); 25.5 +/- 6.4 mm (group C); 32.6 +/- 4.2 mm (group D); 49.4 +/- 5.0 mm (group E). Bipolar RFA with instillation of 6% NaCl solution through an open perfusion system demonstrates better efficacy in creating a larger ablation zone than does conventional or simultaneous monopolar modes at the various times examined. Therefore, hypertonic saline-enhanced bipolar RFA seems to be a promising approach for treating larger liver tumors. PMID- 15789260 TI - Archival study of a Finnish isolate from the 1988/89 rabies outbreak. AB - An archival brain sample from an infected dog taken during the 1988/89 rabies outbreak in Finland was investigated to confirm that it was infected with classical rabies virus (genotype 1). The first 400 base pairs of the virus nucleoprotein gene were then sequenced and used to undertake a phylogenetic comparison between this isolate and a panel of rabies virus sequences from the Baltic region. The Finnish dog isolate was identical to a virus isolated from a raccoon-dog also recorded during the 1988/89 outbreak and closely related to a third isolate from the Russian city of Pskov. The implications of these observations for the possible source of the Finnish outbreak are discussed. PMID- 15789261 TI - HBx protein of hepatitis B virus (HBV) can form complex with mitochondrial HSP60 and HSP70. AB - HBx, a transcriptional transactivating protein of hepatitis B virus (HBV), is required for viral infection and has been implicated in virus-mediated liver oncogenesis. However, the molecular mechanism for its influence on cell remains largely unknown. It was proved that HBx need the help of host cell proteins to exert its function by binding to them. During purifying of GSTX (fusion protein of GST and HBx) expressed in E. coli, we found that it can bind specifically with GrpE (HSP60) and DnaK (HSP70) of E. coli while GST cannot. Using GST pull-down, two-dimensional gel electrophoresis and mass spectrum, we found that GSTX can also bind to human mitochondrial HSP60 and HSP70, which are homologues of GrpE and DnaK. These interactions between HBx and mitochondrial HSP60 and HSP70 are supported by the result of co-immunoprecipitation experiment. It means that HBx can form complex with E. coli and human HSP60 and HSP70. The implication of HBx, HSP60 and HSP70 complex in molecular mechanism of virus infection is discussed. PMID- 15789262 TI - A new potyvirus from Thunberg fritillary (Fritillaria thunbergii Miq.) in Zhejiang, China. AB - A potyvirus causing mosaic symptoms in Thunberg fritillary (Fritillaria thunbergii) was found at two sites in Zhejiang province, China. The virus was readily mechanically transmitted to its original host but not to any of 17 other widely used plant virus indicators. A polyclonal antiserum raised to purified virus particles reacted with its homologous virus but not with a range of other viruses (including 16 potyvirus species). In electron microscopy, virus particles and inclusion bodies typical of a potyvirus were seen. The complete nucleotide sequence of an isolate from Ningbo was determined. It was 9723 nt long and sequence analyses predicted the standard potyvirus organisation. The partial sequence (1664 nts at the 3'-terminus) of an isolate from Panan was also determined; the two sequences had 96.9% nt identity. In sequence comparisons and phylogenetic analyses with completely sequenced potyviruses, the new virus was most closely related to Lily mottle virus (53.0% aa identity) and Leek yellow stripe virus. The most closely related incomplete sequence in the international databases was for Lycoris mild mottle virus (72.8% nt identity in their coat proteins). These results suggest that the virus studied is a new species in the genus Potyvirus, which we have tentatively named Thunberg fritillary mosaic virus. PMID- 15789263 TI - Generation of recombinant norovirus-like particles (VLP) in the human endothelial kidney cell line 293T. AB - Noroviruses (NVs) are the major cause of non-bacterial outbreaks of gastroenteritis. Here we report a new alternative system to generate recombinant NV virus-like particles (rNV-VLP) in a human endothelial kidney cell line (HEK). Transfecting HEK-293T cells with an expression vector coding for the ORF-2 gene lead to the expression of the viral structural protein VP1 which spontaneously assembled into virus-like particles (VLP), as shown by electron microscopy. The transfected cells did not show a cytopathic effect and released rNV-VLP into the culture medium. The HEK-293T cell derived particles were morphologically indistinguishable to the rNV-VLP produced from baculovirus and the Venezuelan equine encephalitis virus (VEE)-replicon. The produced particles were stable for at least 2.5 months at 4 degrees C. PMID- 15789264 TI - Etiologic agents of acute gastroenteritis among Japanese infants and children: virus diversity and genetic analysis of sapovirus. AB - A total of 371 fecal specimens from infants and children with acute gastroenteritis in Maizuru, Tokyo and Osaka, Japan from July 2002 to June 2003 were tested for the presence of diarrheal viruses by reverse transcription polymerase chain reaction (RT-PCR), reverse passive hemagglutination (PRHA), RNA polyacrylamide gel electrophoresis (PAGE), latex agglutination and sequence analysis methods. Among diarrheal viruses detected, group A rotavirus was the most prevalent (42.2%) followed by norovirus (28.9%), group C rotavirus (8.4%), sapovirus (6.7%), adenovirus (5.3%) and astrovirus (0.9%), respectively. There was the high rate (7.6%) of viral mixed infections. Sapovirus was classified into 6 genotypes (GI/1, GI/4, GI/5, GI/6 and GII/1 and one novel tentatively called GII/5). It is noteworthy that genogroup II sapovirus can be classified into 5 genotypes. Our findings confirmed the presence of many diarrheal viruses co circulating among Japanese infants and children and showed the great genetic diversity among sapoviruses. PMID- 15789265 TI - Sequence characterization of tomato leaf curl Sinaloa virus and tomato severe leaf curl virus: phylogeny of New World begomoviruses and detection of recombination. AB - Diseases caused by begomoviruses (family Geminiviridae, genus Begomovirus) constitute a serious constraint to tomato production in Nicaragua. In this study, the complete nucleotide (nt) sequences of the DNA-A and DNA-B components were determined for the first time for Tomato leaf curl Sinaloa virus (ToLCSinV). In addition, the complete nt sequence was determined for the DNA-A component of two isolates of Tomato severe leaf curl virus (ToSLCV). The genome organization of ToLCSinV and ToSLCV was identical to the bipartite genomes of other begomoviruses described from the Americas. A phylogenetic analysis of DNA-A including 45 begomovirus species showed that the indigenous begomoviruses of the New World can be divided into three major clades and an intermediate group: AbMV clade, SLCV clade, "Brazil clade", and BGYMV group. Phylogenetic analyses of the DNA-A and DNA-B components and their open reading frames indicated that ToLCSinV and ToSLCV belong to different clades: ToLCSinV to the AbMV clade, and ToSLCV to the SLCV clade. The two Nicaraguan isolates of ToSLCV showed a close relationship with ToSLCV from Guatemala (ToSLCV-[GT96-1]) and Tomato chino La Paz virus (ToChLPV), but differed significantly in the AV1 and AC1 regions, respectively. Computer based predictions indicated that recombination with another begomovirus had taken place within AV1 of ToSLCV dividing this species into two strains. A high probability was also found that ToChLPV is involved in the evolution of ToSLCV. PMID- 15789266 TI - Genotypic distribution of human herpesvirus-8 strains circulating in HIV-positive patients with and without Kaposi's sarcoma in Hungary. AB - Open reading frame (ORF) 26 of human herpesvirus-8 (HHV-8) from peripheral blood samples of 15 Hungarian HIV-positive patients with or without Kaposi's sarcoma (KS) were amplified and sequenced. Four variants of HHV-8 were identified according to ORF 26 genotyping. Most of the samples were shown to be subtype A3, however, subtypes A, B3/C2/C2', and C3 (ORF 26 region) were also identified. The ORF 26 subtypes A and C3 of HHV-8 were only recovered from patients with KS while A3 was dominant in KS negative cases. The amplification of the hypervariable ORF K1 gene was successful only from 2 of the same 15 patients. Sequence analysis of the amplified ORF K1/VR1 regions identified subtype A3 from 2 patients with AIDS associated KS. A novel ORF K1/VR1 variant belonging to subgroup A' was detected in a different sample in one of them. Amplification of the ORF K15, another representative locus for HHV-8 genotyping, was not successful from any of the peripheral blood samples. Unsuccessful amplification of the terminal K1 and K15 ORFs from peripheral blood samples suggests that KS biopsy specimens are needed for complete genotyping of HHV-8 strains from Hungary. PMID- 15789267 TI - Genomic variability of prune dwarf virus as affected by agricultural practice. AB - Twelve new sequences of the coat protein gene of Prune dwarf virus (PDV) variants, obtained from almond trees, are presented. Comparison with previously reported sequences of the same region, obtained from other hosts (plum, cherry and peach) revealed not only the existence of a wider range of variants of PDV than formerly predicted, but also the frequent presence of a mixture of variants in each sample. In spite of the heterogeneity found in almond, the amino acid composition of the domain at the N terminus of the coat protein maintained the potential to form an amphipathic helix, and hence the capacity to serve the previously suggested function of binding the viral RNA during particle formation. Except for synonymous substitutions, measures of nucleotide diversity calculated for the two groups, respectively 13 sequences from almond and 14 sequences from other hosts, were found to be significantly different, with the almond group showing a much higher variability. Analysis of the dendrogram constructed based in all 27 PDV CP sequences did not reveal host specificity, in agreement with previous findings. However, a clear divergence between almond and other hosts sequences could be found. It is discussed that the observed differences between almond and other hosts variants may derive from differences in agricultural practices. PMID- 15789268 TI - Genome segment RNA-1 of a flat apple isolate of Cherry rasp leaf virus: nucleotide sequence analysis and RT-PCR detection. AB - The sequence of the RNA-1 of a flat apple isolate of Cherry rasp leaf virus (CRLV FA) was determined using overlapping cDNA fragments. CRLV-FA RNA-1 consists of 6992 nucleotides (nt), excluding a 3' poly (A) tail. A single open reading frame (ORF) consisting of 6705 nt was identified. This ORF encodes a putative polyprotein consisting of 2235 amino acid (aa) residues, approximately 249.6 kDa. When compared to CRLV-pot (potato isolate) RNA-1 ORF, 2 deletions of 5 aa and 10 aa (total 15 aa) were observed at the variable N-terminus of the protease cofactor of CRLV-FA. Non-coding regions were identified at the 5'-(142 nt) and 3' end (145 nt). CRLV-FA and CRLV-pot are isolates of the same virus with identity levels for the RNA-1 associated nt and deduced aa of 94% and 95%, respectively. RT-PCR targeting CRLV-FA RNA-1 appear to be of similar sensitivity and just as reliable as RT-PCR targeting RNA-2. PMID- 15789269 TI - Postreassortment changes in a model system: HA-NA adjustment in an H3N2 avian human reassortant influenza virus. AB - In our previous studies we described the postreassortment changes in the hemagglutinin (HA) of H2N1, H3N1, H4N1 and H13N1 influenza A virus reassortants with HAs derived from avian viruses and low-functional neuraminidase (NA) of a human parent virus A/USSR/90/77 (H1N1). The changes involved amino acid substitutions that increased the negative local charge in the vicinity of the receptor-binding pocket and decreased the affinity of HA to sialic acid receptors. In the present report we describe the studies performed with H3N2 reassortant viruses having HA of A/Duck/Ukraine/1/63 (H3N8) virus and NA of A/Aichi/2/68 (H3N2) virus. Amino acid changes in the HA gene registered in virus variants selected in the course of serial passages resulted in a decrease in the affinity to sialic acid-containing substrates and cell receptors. However, the decrease was less expressed than in the reassortants containing the low functional NA of N1 subtype described in our earlier studies, and the amino acid changes were not necessarily associated with an increase of negative charge. In one passage variant an amino acid substitution in NA was detected. The relevance of these results for the evolution of the H3N2 virus of the 1968 pandemic is discussed. PMID- 15789270 TI - Molecular characterization of a virus from the family Luteoviridae associated with cotton blue disease. AB - Cotton blue disease is an aphid-transmitted cotton disease described in Brazil in 1962 as Vein Mosaic "var. Ribeirao Bonito". At present it causes economically important losses in cotton crops if control measures are not implemented. The observed symptoms and mode of transmission have prompted researchers to speculate that cotton blue disease could be attributed to a member of the family Luteoviridae, but there was no molecular evidence supporting this hypothesis. We have amplified part of the genome of a virus associated with this disease using degenerate primers for members of the family Luteoviridae. Sequence analysis of the entire capsid and a partial RdRp revealed a virus probably belonging to the genus Polerovirus. Based on our results we propose that cotton blue disease is associated with a virus with the putative name Cotton leafroll dwarf virus (CLRDV). PMID- 15789271 TI - Leptin stimulates the reproductive male axis in rats during sexual maturation by acting on hypothalamic excitatory amino acids. AB - The purpose of the present study was to determine the effect of treatment with leptin on gonadotrophin secretion and hypothalamic GnRH, excitatory and inhibitory amino acids release, in prepubertal (15 days old) and peripubertal (30 days old) male rats. Rats of both ages received a single (ip) injection of 30 microg/kg leptin 60 minutes previous to sacrifice. Serum LH was determined, and the hypothalamus dissected and incubated in Earle's medium. GnRH and amino acids release were determined in the media. LH and GnRH were measured by RIA. Amino acids were assessed by HPLC-UV detection. In the two prepubertal stages, (prepubertal and peripubertal, 15 and 30 days of age respectively) leptin increased plasmatic LH levels (p < 0.01) and hypothalamic GnRH release (p < 0.01). Glutamate (GLU) release showed an increment in leptin-treated rats (p < 0.01) at both ages, while only the 30 days old rats showed an increment of the aspartate (ASP) release. GABA secretion was not modified by leptin treatment. In conclusion, the results demonstrated that leptin stimulates the LH-GnRH axis during sexual development in male rats, increasing the secretion of both hormones. The hypothalamic excitatory amino acid neurotransmitter system appears to be involved in this change. PMID- 15789272 TI - Four-year follow-up of acromegalic patients treated with the new long-acting formulation of Lanreotide (Lanreotide Autogel). AB - Lanreotide Autogel (Ipsen) is a long-acting somatostatin analogue (SA) in a new galenic formulation suitable for subcutaneous (s.c.) injection. In our department, 11 patients with therapy-resistant acromegaly were treated with Lanreotide Autogel for 48 months. 10/11 patients had previously undergone transsphenoidal surgery. For a median duration of 1.4 years prior to Lanreotide Autogel, the patients received Lanreotide PR 30 mg every 7, 10, or 14 days. 60, 90, or 120 mg of Lanreotide Autogel was administered by deep s.c. injection every 28 days, with the higher dosage being given to those with the previously shortest injection interval under Lanreotide PR. Dose was adjusted on the basis of Growth Hormone (GH) level after 4, 8, and 12 months with a minimum dose of 60 mg and a maximum dose of 120 mg. The efficacy of Lanreotide Autogel treatment was evaluated by measuring GH concentrations (4 hour profiles) and IGF-I levels. Before switching to Lanreotide Autogel, the multiple of the upper limit of normal (xULN) of IGF-I levels was 1.2 (median) and the median GH level was 1.3 microg/l. 3 out of 11 patients had an IGF-I within the age- and sex-adjusted normal range. After 48 months of treatment with Lanreotide Autogel, six patients had an IGF-I within the normal range. Median GH levels were at 1.3 microg/l and xULN of IGF-I was at 1.0 compared to Lanreotide PR 30 mg treatment (p < 0.001). At the end of the study, 8 patients received 120 mg Lanreotide Autogel, 2 patients 90 mg and 1 patient 60 mg, respectively. There was slight but significant deterioration of glucose metabolism with an increase of HbA1c. In conclusion, the new galenic formulation of Lanreotide improves not only the control of biochemical markers of acromegaly compared to the conventional PR formulation, but is also easier to administer given its deep s.c. method of administration. Glucose metabolism has to be followed carefully in patients on high-dose Lanreotide Autogel. PMID- 15789273 TI - Amiodarone-induced thyrotoxicosis due to destructive thyroiditis: therapeutic recommendations. AB - BACKGROUND: Amiodarone-induced thyrotoxicosis (AIT) type 1 occurs in subjects with an underlying thyroid disease, whereas type 2 AIT is a form of destructive thyroiditis. Our hypothesis was that the common practice of thyroid testing before prescription of amiodarone would reduce the incidence of pure type 1 AIT, though a stringent classification may be difficult (mixed type AIT). MATERIALS AND METHODS: Thyroid testing before and after treatment of AIT (n = 12) and the response to combined antithyroid and glucocorticoid treatment (n = 11) were recorded in a consecutive series of patients seen at a university hospital. RESULTS: Some criteria for type 1 AIT were fulfilled in 3 patients, but the diagnosis of a mixed form AIT was more likely in 2 of these. Type 2 AIT was diagnosed in the other 9 patients, while 6 patients had diffuse hypoechoic goitre. The median time to euthyroidism (defined as normal fT3 concentration) under thionamide and prednisolone (starting dose 20 to 75 mg/d) was 2 months (interquartile range 1 to 2.7 months). Thionamide treatment was stopped after a median duration of 5.7 months (interquartile range 4.2 to 8.7 months) and glucocorticoids were completely withdrawn after 6.7 months (5.5 to 8.7 months). CONCLUSIONS: Nowadays, isolated type 1 AIT is rarely found and destructive thyroiditis (as type 2 AIT or mixed form) is the predominant cause of AIT. To accelerate recovery, we prescribed thionamide and glucocorticoids simultaneously as first-line therapy once contraindications for the use of steroids had been ruled out. PMID- 15789274 TI - Questionnaire on management of nodular thyroid disease (Annual Meeting of the Thyroid Section of the German Society of Endocrinology 2003). AB - Nodular thyroid disease is highly prevalent in iodine deficient areas. In Germany it affects approximately 30 % of the adult population. Differential diagnosis of thyroid nodules is directed at exclusion of rare thyroid malignancy and assessment of the nodules' functional characteristics in order to determine the best treatment approach. In 2003 the annual meeting of the Sektion Schilddruse, a thyroid specialist group of the German Society of Endocrinology addressed the topic of "management of benign nodular thyroid disease". To assess the current diagnostic and therapeutic approach to nodular thyroid disease by German thyroid experts we designed a questionnaire, which was sent to all members of the Thyroid Section before the meeting and was answered by 70 % of the participants. We here report the results of the questionnaire, which show a considerable national heterogeneity in the management of thyroid nodules as well as diagnostic and treatment preferences for different thyroid specialist subgroups, e.g. endocrinologists and nuclear medicine specialist. This is in agreement with results of previous questionnaires assessing state-of-the-art practise among members of the European and American Thyroid Associations and underlines the definite need for a consensus process and for carefully planned randomized trials to answer the many unresolved questions in diagnosis and therapy of nodular thyroid disease. PMID- 15789275 TI - Perinatal morbidity in women with undiagnosed gestational diabetes in northern thuringia in Germany. AB - INTRODUCTION: Undiagnosed gestational diabetes mellitus (GDM) is associated with severe perinatal complications. PATIENTS AND METHODS: Out of 970 women, infant and maternal morbidity was assessed in 114 mother-children-pairs with an infant birth weight over the 90th percentile (Voigt et al., 1996). It was the aim of this retrospective study to assess the number of mothers with undiagnosed GDM, who have born a macrosomic child. RESULTS: The macrosomia rate in newborns was 12 % in this study excluding macrosomic infants of mothers with preexisting diabetes mellitus. Maternal data: Age 28.3 +/- 5.3 years, adipositas (body-mass-index > 30 kg/m) in 42.1 % vs. 30.4 % in the peer group (p < 0.02), increase in weight > 15 kg during pregnancy in 57.9 % of the mothers who have given birth to a macrosomic child vs. 30.9 % in the peer group (p < 0.0001), family history of diabetes mellitus (28.0 % vs. 11.3 % in the peer group, p = 0.006), preeclampsia in 8.8 % vs. 2.7 % in the peer group (p = 0.002), cervical insufficiency in 2.6 % vs. 0.4 % in the peer group (p = 0.02). After delivery HbA1c was elevated in 38.6 % of the women having born macrosomic infant (mean HbA1c: 5.0 % +/- 0.5). Infant data: neonatal jaundice 16.7 % vs. 4.5 % in the peer group, p < 0.0001. There were no statistically significant differences concerning perinatal condition and malformations. Neonatal hypoglycaemia occurred in 9.6 % of the macrosomic infants. Cord blood insulin levels were significantly elevated in comparison to the peer group of mothers without metabolic disorders and having born eutroph infants (8.4 mU/l [3.0 - 100.0] vs. 5.3 mU/l [3.0 - 30.7], p = 0.01). 11.4 % of all macrosomic infants had cord blood insulin levels above the normal range. CONCLUSION: More than one third of the mothers having born one or more macrosomic infants had an impairment of glucose metabolism immediately after birth. The elevated prevalence of preeclampsia in this group confirms the relationship of hypertension and impaired glucose metabolism during pregnancy. The detection of hyperinsulinaemia, postnatal hypoglycaemia, elevated prevalence of neonatal jaundice with need of further therapy and diabetic fetopathy in macrosomic infants of mothers, whose metabolism was not monitored during pregnancy, pinpoint the need for a diagnostic screening for GDM. PMID- 15789276 TI - Clinical characteristics of type 2 diabetes mellitus in overweight European caucasian adolescents. AB - AIM: To present the clinical features of Type 2 diabetes mellitus (T2DM) in overweight European Caucasian children and adolescents. METHODS: We report the clinical characteristics of 16 non-syndromal overweight European Caucasian adolescents with T2DM (10 boys, 6 girls, SDS-BMI in median +2.8, range +1.6 to +3.4) treated in 5 specialised centres for obesity and diabetes. RESULTS: None of the adolescents manifested with ketoacidosis. 13 were asymptomatic (3 adolescents with polyuria), 12 showed features of metabolic syndrome (dyslipidaemia or hypertension), 8 demonstrated acanthosis nigricans and 12 had relatives with T2DM. 11 adolescents were extremely obese and all patients were pubertal. Mean age at diagnosis was 14.2 years (range 11.0 - 16.9). Median insulin was 19 microU/ml, insulin resistance index (HOMA) 8.5, C-peptide 2.3 ng/ml, HbA1c 6.9 %, fasting blood glucose 176 mg/dl and blood glucose at 2 hours with the oGTT 229 mg/dl at manifestation. Fasting blood glucose and HBA1c were in the normal range in 4 and 6 adolescents respectively, while oGTT always fitted the diagnosis of T2DM. CONCLUSION: Since T2DM occurred in Caucasian overweight adolescents and is frequently asymptomatic, it is essential that clinicians perform diagnostic procedures to identify T2DM in high-risk groups of overweight Caucasian adolescents (extreme obesity, features of metabolic syndrome, relatives with T2DM). PMID- 15789277 TI - Acute hyperinsulinemia is associated with increased plasma adrenomedullin concentrations in uncomplicated obesity. AB - OBJECTIVE: Adrenomedullin (AM) is a potent hypotensive peptide which may be implicated in the insulin regulatory system. Acute hyperinsulinemia exerts no influence on plasma AM in normal subjects while no data on obese subjects has been reported. PURPOSE: The aim of the study was to investigate the effect of acute hyperinsulinemia on the plasma AM concentration in patients with uncomplicated obesity. RESEARCH METHODS: We measured the plasma AM levels in 23 obese subjects (BMI 41.9 +/- 9.8 kg/m2), 21 females and 2 males (mean age 31 +/- 7.2 years), before and during a euglycemic hyperinsulinemic clamp. The control group consisted of 43 healthy subjects (HS) (22 males and 21 females; mean age 38 +/- 12 years; BMI 23.3 +/- 3.2 kg/m2). RESULTS: Baseline plasma AM was found to be higher in obese subjects (20.4 +/- 8.4 pg/ml) than in normal subjects (11.3 +/ 0.8 pg/ml) (p < 0.001). A significant increase in the plasma AM levels was observed in obese subjects during acute hyperinsulinemia (from 20.4 +/- 8.4 pg/ml at 0 min to 26 +/- 8.9 pg/ml at 120 min, p < 0.02). Plasma AM concentrations were significantly correlated with insulin levels at 30 min (r = 0.44; p = 0.04) and 120 min (r = 0.40, p = 0.05) during the clamp. DISCUSSION: In conclusion, acute hyperinsulinemia induced a significant increase in the plasma levels of AM in uncomplicated obese subjects. Hyperinsulinemia may, at least in part, regulate levels of AM in obesity, explaining the high levels of the peptide in these subjects. PMID- 15789278 TI - Biphasic insulin aspart compared to biphasic human insulin reduces postprandial hyperlipidemia in patients with Type 2 diabetes. AB - BACKGROUND: Premixed insulin analogues reduce postprandial hyperglycemia in patients with Type 2 diabetes in comparison to premixed regular insulin. Insulin also plays an important role in the regulation of postprandial lipid metabolism. It is known that increased levels of postprandial insulin reduce postprandial hyperlipemia but, on the other hand, no information exists with regard to the possible effect of insulin analogues in comparison to human insulin. MATERIALS AND METHODS: 12 subjects (3 men; age 59 +/- 5 years; BMI 30.5 +/- 5.9 kg/m2, duration of diabetes 9 +/- 1 years, HbA1c 8.33 +/- 1.1 %) already on therapy with premixed insulin were treated either with biphasic human insulin (BHI30) or with biphasic insulin aspart (BIAsp30) (1.3 IU fast acting insulin/12 g KH) in the setting of a standardized test meal. Serum levels of glucose, insulin, C-peptide and triglycerides as well as retinylpalmitate in plasma and chylomicron remnants were determined before and up to 8 hours after the meal. RESULTS: As was to be expected, therapy with BIAsp30 reduced the maximum increase of postprandial glucose from 7.10 +/- 2.00 mmol/l to 5.27 +/- 1.83 mmo/l (p = 0.007) compared to BHI30 insulin. In the same way, the maximum increase of triglycerides (from 2.33 +/- 1.03 to 1.65 +/- 0.69 mmol/l, p = 0.014) was reduced. The AUC 0 - 8 for triglycerides was not significantly influenced (34.20 +/- 19.86 vs. 31.46 +/- 16.21 mmol x 8 h/l) but the incremental area over baseline (AOB 0 - 8) was significantly reduced from 8.02 +/- 4.35 to 6.12 +/- 3.94 mmol x 8 h/l (p = 0.024). CONCLUSIONS: Compared to conventional human premixed insulin the prandial therapy with biphasic insulin aspart results not only in an improvement of glucose tolerance but also in a significant reduction of postprandial hyperlipemia. PMID- 15789279 TI - The recombinant rat glucagon-like peptide-1 receptor, expressed in an alpha-cell line, is coupled to adenylyl cyclase activation and intracellular calcium release. AB - The glucagon-like peptide-1 (GLP-1) receptor is expressed on alpha-cells, though its functional significance is unknown. The endogenous beta-cell GLP-1 receptor is coupled to adenylyl cyclase, cell depolarization, activation of voltage dependent Ca2+ channels (VDCC) and extracellular Ca2+ influx (Lu et al., 1993 b). In contrast, the signaling pathways of the GLP-1 receptor in alpha-cells are poorly understood. To determine the signaling mechanisms of the alpha-cell GLP-1 receptor, we established a stable pancreatic islet alpha-cell line expressing the recombinant rat GLP-1 receptor (INR1-SF2), using INRl-G9 cells. These INRl-G9 cells do not express endogenous GLP-1 receptor. In INR1-SF2 cells, GLP-1 bound to the recombinant receptor (Kd = 0.9 nM) and increased cAMP (ED50 = 0.6 nM). GLP-1 increased the free cytosolic Ca2+ ([Ca2+]i) (ED50 = 50 nM) by release from intracellular stores, but did not affect INR1-SF2 cell phosphoinositol turnover. Despite expressing VDCC, the INR1-SF2 cells were not depolarized by GLP-1, even in the presence of glucose. This contrasts with the depolarizing action of GLP-1 in beta-cells in the presence of glucose (Lu et al., 1993 b). This study establishes that a single GLP-1 receptor species can mediate the effects of GLP-1 through multiple signaling pathways, including the adenylyl cyclase system and intracellular Ca2+ release, in an alpha-cell type. Furthermore, since GLP-1 is unable to cause cellular depolarization or activate VDCC in INR1-SF2 cells, these data suggest that glucose-induced membrane depolarization may be crucial for GLP 1 to further activate VDCC and potentiate glucose-stimulated insulin release in beta-cells. Finally this study describes a cell line that can be used as a model system for evaluation of GLP-1 signaling in alpha-cells. PMID- 15789280 TI - [Health Status of the German population: results of a representative survey using the EuroQol questionnaire]. AB - PURPOSE: The EuroQol questionnaire (EQ-5D) is an instrument for subjectively describing and valuing health states. The purpose of this study was to measure the health status of the German population using the EQ-5D and to analyse the influence of sociodemographic factors. METHODS: As part of the European Study of the Epidemiology of Mental Disorders (ESEMeD) a representative sample of the non institutionalised population aged 18 and above in Germany was surveyed by personal computer-based interviews in 2002/2003. Besides questions relating to the prevalence of psychiatric disorders and their risk factors, the health status of 3552 respondents was recorded using the EQ-5D. Frequency and factors of influence on problems in the EuroQol items mobility, self-care, usual activities, pain/discomfort, anxiety/depression as well as the valuation of health state on the visual analog scale (VAS) were analysed. RESULTS: 36 % of the respondents report problems in at least one of the EuroQol items; most frequent were problems in the dimension pain/discomfort (28 %), followed by mobility (17 %), everyday activities (10 %), anxiety/depression (4 %) and self-care (3 %). The mean VAS score was 77.4. Except for anxiety/depression the frequency of problems increased significantly with age and decreased with income. Women reported significantly more problems than men with regard to pain/discomfort and self-care. Having received more than 9 years of school education and being employed was associated with significantly fewer problems in all dimensions of the EuroQol. Living with a partner was associated with significantly fewer problems in the dimensions of self-care, usual activities and anxiety/depression. Multivariate analyses confirmed the effect of age, school education and employment on the frequency of problems, whereas for living with a partner only effects on self-care and anxiety/depression, and for gender only effects on pain/discomfort were confirmed. Income had no effect on frequency of problems in multivariate analyses. Young age, more than 9 years of school education, employment and living with a partner had a significantly positive effect on VAS score in multivariate analyses, while gender and income had no effect. CONCLUSIONS: The EQ-5D is a simple health state classification system, that can be used to detect variations in the health status of a population. This study provides EQ-5D-norm values for the German population. PMID- 15789281 TI - [Early factors influencing body weight and prevalence of overweight in 4610 children prior to school entry in the Erlangen District (Northern Bavaria)]. AB - OBJECTIVE: Childhood obesity is a growing health problem among German children. Birth weight is considered to have a major influence on the course of postnatal weight. The present study investigates the relation between birth weight and body weight at health examination prior to school entry. We tested other parameters influencing childhood obesity, and hypothesized that within a 5-year interval the prevalence of obesity in children has increased significantly. METHODS: Our study is based on the retrospective analysis of the school enrolment health examinations of 4610 children in North Bavaria, e. g. the town Erlangen and administrative district Erlangen-Hoechstadt in 1995/96 and 2000/01. RESULTS: A higher birth weight was associated with a higher weight and BMI at school entry examination (p < 0.0001). An increased birth weight is therefore a considerable risk factor for later overweight in childhood. Hypotrophic newborn, however, gain less weight and exhibit a lower BMI in our study group. In general, boys were significantly heavier than girls (p < 0.001). Children of foreign origin were heavier and had a higher BMI corrected for age than German pupils but they were also 0.07 years older. Our regional survey revealed local differences in the prevalence of obesity. Comparing the cohorts 1995/96 and 2000/01 at school entry, a significant increase of BMI in the latter was found (p < 0.0001). CONCLUSIONS: A highly significant increase in the prevalence of infantile overweight has to be faced. Early prevention of childhood obesity is therefore mandatory to avoid the complications and higher morbidity. PMID- 15789282 TI - [Ratio of confirmed versus suspected occupational diseases as a parameter of quality]. AB - PURPOSE: To evaluate the notifications of suspected causes of occupational diseases (NSC) under quality assurance aspects. METHODS: We retrospectively analysed the NSC assessed by the institutions for statutory accident insurance and prevention (Berufsgenossenschaft) between 1998 and 2002. RESULTS: Physicians were most frequently represented among the 357,050 notifications (60.2 %). The number of notifications dropped continually between 1998 and 2002. The number and quality of the notifications (NSC) depend on the knowledge of the occupational diseases as well as on the knowledge of the insured persons and on their respective occupational histories. The NSC are issued via the statutory social insurance bodies who are in contact with the patients, and the NSC also depend on the present scientific knowledge of the types of relevant diseases. Usually, the physicians were most frequently the notifiers of occupational diseases, with one exception, namely, incapacitating diseases due to damage to vertebral discs of the lumbar spine caused by many years of lifting of carrying heavy loads or of many years of work in an extremely bent position, in which cases the statutory insurance bodies themselves issued the notifications. Altogether 123,262 notifications were confirmed. The average quota of confirmations was > 40 % with physicians issuing the notifications, > 30 % with statutory pension insurance bodies and only approx. 12 % with notifying statutory sickness insurance bodies. The lowest quota of confirmations - namely, only 3 % - concerned the incapacitating vertebral disc disease of the lumbar spine mentioned above. Non confirmed notifications of suspected occupational diseases involved an estimated cost of at least 58 million Euro. DISCUSSION: On-target improvement of the quality of notification of suspected occupational diseases is possible by exploring several avenues, including on-target assessment of evaluation criteria reporting by the statutory accident insurance bodies concerned and by regular advice rendered to the statutory social insurance bodies by occupational therapists. PMID- 15789283 TI - [Effects of the new German quarterly practice fee on dental treatment -- cost benefit analysis]. AB - BACKGROUND: In the German Law on the Modernization of the Statutory Healthcare System of 19 November 2003 the legislator enacted payment of a so-called quarterly practice fee for ambulant medical and dental care. The legislator hopes that by the introduction of this fee the central "steering" function of the family doctor is strengthened and that medically unnecessary treatment will be reduced, thus reducing cost and consolidating health insurance funds. In public, however, the introduction of the practice fee as a steering instrument is very controversially discussed. The aim of the analysis is to balance the cost effectiveness of a quarterly practice fee with regard to the specific features of the dental care system. RESULTS: Analysis of the allocative and distributive effects shows that the practice fee does not adequately take into account the specific features of the dental care sector and that the adverse effects prevail if seen under the aspect of health economics and social medicine. Neither on the micro-economic nor on the macro-economic level a noticeable medicine-term or long term cost containment effect may be expected. PMID- 15789284 TI - [Amalgam risk assessment with coverage of references up to 2005]. AB - Amalgam, which has been in use in dentistry for 150 years, consists of 50 % elemental mercury and a mixture of silver, tin, copper and zinc. Minute amounts of mercury vapour are released continuously from amalgam. Amalgam contributes substantially to human mercury load. Mercury accumulates in some organs, particularly in the brain, where it can bind to protein more tightly than other heavy metals (e. g. lead, cadmium). Therefore, the elimination half time is assumed to be up to 1 - 18 years in the brain and bones. Mercury is assumed to be one of the most toxic non-radioactive elements. There are pointers to show that mercury vapour is more neurotoxic than methyl-mercury in fish. Review of recent literature suggests that mercury from dental amalgam may lead to nephrotoxicity, neurobehavioural changes, autoimmunity, oxidative stress, autism, skin and mucosa alterations or non-specific symptoms and complaints. The development of Alzheimer's disease or multiple sclerosis has also been linked to low-dose mercury exposure. There may be individual genetical or acquired susceptibilities for negative effects from dental amalgam. Mercury levels in the blood, urine or other biomarkers do not reflect the mercury load in critical organs. Some studies regarding dental amalgam reveal substantial methodical flaws. Removal of dental amalgam leads to permanent improvement of various chronic complaints in a relevant number of patients in various trials. Summing up, available data suggests that dental amalgam is an unsuitable material for medical, occupational and ecological reasons. PMID- 15789285 TI - [Guidelines and recommendations for ensuring Good Epidemiological Practice (GEP) - revised version after evaluation]. PMID- 15789288 TI - [Development and psychometric testing of a patient questionnaire for medical rehabilitation (IRES-3)]. AB - BACKGROUND: Re-analyses of extensive datasets as well as theoretical considerations have led to the conclusion that the patient questionnaire "Indicators of Rehabilitation Status" (IRES) should be revised in several respects. The new version IRES-3 was developed on the basis of a theoretical model of rehabilitation following the ICF (International Classification of Functioning, Disability and Health) as well as of elements of the earlier version IRES-2. In addition, we included the results of expert panels on the definition of treatment goals in medical rehabilitation. METHODS: The IRES-3 was tested in 453 patients in rehabilitation clinics of seven diagnostic areas. To compare the IRES-3 with other generic instruments, the SF-36 and the HADS were employed at the same time. For purposes of cross-validation, an external sample could be used. The dimensional structure was tested in confirmatory factor analyses. RESULTS: With rare exceptions, the tests showed good values for item difficulties, ceiling and floor effects, internal consistencies and test-retest reliability. Convergent validity could be established for the IRES-3 when compared to relevant scales of the SF-36 and the HADS. Indices of sensitivity to change were comparable, if not somewhat superior to the effects on comparable scales of the SF-36. The assessment of the construct validity and the dimensional structure of the questionnaire led to the definition of eight dimensions which can be interpreted as somatic, emotional, functional, occupational, social, pain, coping, as well as health information and behaviour. The dimensional structure of the IRES-3 can be regarded as theoretically meaningful as well as empirically proven. STANDARDIZATION: For purposes of standardization, data were collected on a sample representative of the population aged 30-75 years in Germany. These data allow a norm-oriented interpretation of scales and dimensions of the IRES-3 for patients in rehabilitation. APPLICATION: The questionnaire is ready for application, including a computer programme for data entry and analysis. PMID- 15789289 TI - [Modification of the subjective prognosis of gainful employment during inpatient rehabilitation. Findings of a study of 180 orthopaedic patients]. AB - Modifications of the subjective prognosis of gainful employment during in-patient rehabilitation were assessed by means of a 3-item scale (SPE scale) in 180 orthopaedic patients. Statistically significant improvements were found as to the subjective prognosis of gainful employment and other measures of vocational ability as well. Patients showing lower risk at the beginning profited to a greater degree from rehabilitation as compared to patients with higher scores in SPE scale. To improve the effects of rehabilitation for patients with a high subjective risk as to gainful employment specific interventions are recommended. Further research is discussed. PMID- 15789290 TI - [Residential rehabilitation]. AB - In recent years new avenues of medical rehabilitation have been made possible in order to allow a more flexible and better care for patients. This includes day care and outpatient treatment additional to inpatient services. A further new development are centers which provide "residential rehabilitation". It allows a continuous cooperation with physicians and therapists who send patients or take care of them after discharge. Cooperation with acute care hospitals is easier. Inpatient care, day care and outpatient care can be better integrated. The social network of the patient can easier be taken into account in treatment planning. Patients can be supported when going back to work. Contact with self-help groups can be arranged while the patient is still in treatment. PMID- 15789291 TI - [Obesity treatment in rehabilitation clinics--explanations and remarks on implementing clinical practice guideline]. AB - This paper focuses on the improvement of obesity treatment during in-patient rehabilitation. ASSIGNMENT: The diagnosis of obesity rarely is in the foreground of assignment to rehabilitation. Obesity is the primary diagnosis in only about 1 % of all adipose patients; the term is also non-specific with respect to the causal role in the Metabolic Syndrome. The effects of obesity (diabetes, hypertension, myocardial infarction, arthrosis) are treated but not the underlying cause. THERAPY: Treatment of obesity can be the primary focus or take place as an adjunct to the treatment of other diseases; it therefore requires different structures and a sufficient number of patients. Life style change is an essential aspect of obesity-specific behavior therapy and requires closed groups. Different modular therapeutic offerings are more suitable in physiotherapy and sports. In general, 85 - 90 % of all patients meet the prerequisites for group settings (i. e., ability to communicate, discipline). A generally accepted uniform pattern of motivational assessment appears to be important to treatment success but has to be postulated as yet. Pre-assessment interviews and subjects' response to proposed therapy settings are useful means of identifying motivated patients. QUALITY: Setting down rehabilitative goals in writing facilitates patients' achieving therapeutic targets. Medical discharge summaries should reflect the process state (motivation, therapeutic modules, concomitant illnesses, target agreements, subjective and somatic changes in quality of life). Ongoing ambulatory care seems important but can rarely be implemented. Accepted indicators of the quality of rehabilitative obesity treatment need to be determined. PMID- 15789292 TI - [Rules on taking medical information from spinal cord injury patients--a proposal]. AB - If medical information of persons with spinal cord injuries is seen more as a useful component of medical care with high responsibility than an inconvenient duty, it can be an important contribution toward rehabilitation. Based upon psychological principles of communication, practical experience and the outcome of the specialized literature, it is possible to show concrete conditions for competent medical conversation in case of delivering bad news. Taking into consideration the suggested behavioural rules that describe how to proceed the informing can become a fruitful element in relationship between medical doctor and patient. PMID- 15789293 TI - [Sensitivity to change]. AB - In rehabilitation research patient questionnaires are widely used for evaluative purposes, i. e. to measure improvements or deteriorations over time. This is only possible if the questionnaires applied appropriately reflect "true" change over time, i. e. they have to be sensitive to change. The aim of this paper is to point out the importance of the "sensitivity to change" concept for evaluative assessment tools and evaluative studies, respectively, considering quality of life research as an example. Various qualitative aspects, e. g. scaling of response options of assessment tools, are covered as well as quantitative methods, i. e. study designs and indices. Furthermore, recommendations for interpretation are given. PMID- 15789294 TI - [Chances and successes of vocational training of disabled youth in occupational and vocational school--findings of the BAR-pilot study "Regional Networks for Vocational Rehabilitation of (Learning-) Disabled Juvenile (REGINE)"]. AB - The pilot study "REGINE" was realized on initiative of the Federal Rehabilitation Council (BAR) to test the opportunities of vocational training of learning disabled youth under "normal" conditions: That means the young people are trained in firms and regular vocational schools, and -- while doing so -- are supported by educational institutions. The project was finished successfully. The results of the evaluation, realized by the Institute for Empirical Sociology at the University of Erlangen-Nurnberg were already published in December 2003. They encourage making use of the opportunities of in-firm vocational training of disabled youth more frequently. More than half of the participants of the first REGINE-cohort successfully finished their vocational training. Nearly 40 % were taken over by the firm that provided vocationally trained, and 12.8 % found a job in another company. The second cohort showed even better results: the corresponding rates are 47.1 and 32.4 %. This success speaks in favour of this place of vocational training, particularly considering the difficult job situation. It seems to be possible, that disabled youth are successfully and cost effectively trained vocationally, if an individual support of both youth and firms can be provided. Prerequisite for this is a conscientious preparation of the vocational training, which may already begin in the last classes of school. "Achilles heel" of the new place of vocational training for disabled youth is the theoretical training in regular vocational schools which usually can not meet the needs of the learning-disabled. These shortcomings had to be compensated by special educational institutions which are not financed by educational administration but by employment agencies (which actually have no jurisdiction over this kind of duties). PMID- 15789295 TI - [Brain damage--what to do? Chances for reform in neurological medical rehabilitation--introduction to the interactive DVfR contribution for the forum "Brain Impact" in context of RehaCare Fair 2004]. PMID- 15789296 TI - [Memorandum on salient oints regarding progressing development of the German Social Legislation Code IX by the Coalition Working Group "Humans Afflicted with Impediments"]. PMID- 15789297 TI - [Report of the Annual Convention of the Lower Saxony/Bremen Rehabilitation Research Network, on the theme "Rehabilitation: barriers, costs, and optimisation", November 19, 2004 in Oldenburg]. PMID- 15789298 TI - [An EU project targets a new european quality standard in the low-vision range]. PMID- 15789300 TI - [BENEFIT Kidney--significance of a nephrology screening at intervention outset and therapy success]. AB - BACKGROUND AND OBJECTIVE: Early specialist care of patients with renal disease, including timely and planned onset of dialysis, determine the course of the disease, quality of life, hospitalization and life expectancy. A multi-centre enquiry by standardized questionnaire was undertaken to define and analyse medical care of newly dialysis-requiring patients. PATIENTS AND METHODS: Data on 551 patients in five different regions of Germany who for the first time required renal replacement treatment were prospectively collected between July 2002 and March 2003. Documentation of history, clinical findings and biochemical tests was done on consecutive patients with a standardized questionnaire, until the desired number of cases was reached. RESULTS: The mean age of the patients (55.4% males) was 64.8 years. 30.7% had diabetes mellitus, 22.3% arterial hypertension/nephrosclerosis and 16.9% glomerulonephritis/vasculitis. Early predominantly nephrological care had been undertaken in 38.7% of patients. 59.0% were cared for almost exclusively by their general practitioner until the time when dialysis was started. 229 patients (41.6%) were referred to specialist (nephrologists) only when dialysis had become necessary. The onset of dialysis was at the right time in only 50.5% of this group. Comparing the care given by nephrologists with that by general practitioners, elective (i.e. planned) dialysis was begun in 81.0% vs. 48.0% (p<0.05). Hospitalization in the two groups was 54.5% vs. 83.7% (p<0.05), the duration of hospital stay 11.4 vs. 17.4 days (p<0.05). CONCLUSION: Fewer than 40% of patients with chronic renal disease in preterminal renal failure (stage IV) were under the care of nephrologists. The lower the degree of nephrological care the more frequent was there a delay in the onset of dialysis treatment. The incidence and the duration of hospital stay was longer. Structured treatment pathways and incentives need to be formulated to reduce the incidence of wrong or substandard treatment of patients with impaired renal function. PMID- 15789301 TI - [A rare case of birenal malacoplakia with renal failure]. AB - HISTORY AND ADMISSION FINDINGS: A 45 year old man was admitted to our hospital because of fever, loss of appetite, and deterioration of general health. For two weeks the patient suffered from diarrhea which had resulted in moderate volume depletion. In addition, he complained of bilateral flank pain at the time of admission. Furthermore, the patient had a history of heavily drinking alcohol as well as cigarette smoking for many years. He had never attended a medical doctor before. INVESTIGATIONS: The patient presented with the clinical picture of acute renal failure and urosepticaemia which was caused by Escherichia coli. The kidneys were found to be at the upper limit of normal by sonography. Magnetic resonance imaging revealed signal-alterations in both kidneys with hyper- and hypointense zones in the renal parenchyma. DIAGNOSIS: To clarify the cause of rapid deterioration of renal function, we performed a renal biopsy. The histology of the renal specimen revealed an unusual type acute bacterial interstitial nephritis most likely due to an infection with E. coli. The clinical picture, the laboratory findings and renal histology, lead to the diagnosis of birenal malakoplakia. TREATMENT AND COURSE: After intravenous and subsequent oral antibiotic therapy the fever and the clinical signs of urosepticaemia subsided and renal function gradually improved. Antibiotic therapy and supplementation with vitamins were continued for 20 weeks. Five years after initial diagnosis, renal function was stable at a glomerular filtration rate of approximately 45 ml/min. CONCLUSIONS: Malakoplakia of the kidney is a rare form of bacterial interstitial nephritis and requires long-term antibiotic therapy. PMID- 15789302 TI - [Colon cancer: molecular pathogenesis and clinical relevance]. PMID- 15789303 TI - [Bronchial asthma versus chronic obstructive lung disease (COPD). From pathogenesis to differential diagnosis and differential therapy]. PMID- 15789304 TI - [Innovation instead of imitation -- news in drug therapy]. PMID- 15789305 TI - [Hormone therapy and menopause]. PMID- 15789306 TI - A note on exact tests of Hardy-Weinberg equilibrium. AB - Deviations from Hardy-Weinberg equilibrium (HWE) can indicate inbreeding, population stratification, and even problems in genotyping. In samples of affected individuals, these deviations can also provide evidence for association. Tests of HWE are commonly performed using a simple chi2 goodness-of-fit test. We show that this chi2 test can have inflated type I error rates, even in relatively large samples (e.g., samples of 1,000 individuals that include approximately 100 copies of the minor allele). On the basis of previous work, we describe exact tests of HWE together with efficient computational methods for their implementation. Our methods adequately control type I error in large and small samples and are computationally efficient. They have been implemented in freely available code that will be useful for quality assessment of genotype data and for the detection of genetic association or population stratification in very large data sets. PMID- 15789311 TI - Can you hear me now? Listening to our patients and ourselves. PMID- 15789312 TI - Estimating third molar extraction difficulty: a comparison of subjective and objective factors. AB - PURPOSE: The purpose of this study was to compare and contrast subjective and objective assessments of variables associated with third molar (M3) extraction difficulty. MATERIALS AND METHODS: To address the research purpose we implemented a prospective cohort study and enrolled a sample of surgeons removing M3s in an ambulatory care setting. Predictor variables were categorized as demographic, anatomic, or operative. The outcome variables were subjective and objective rankings of the importance of the variables in terms of estimating M3 extraction difficulty. Subjective rankings were made by surveying the surgeons and asking them to rank each variable's importance on a scale ranging from 0 (not important) to 100 (extremely important). Objective rankings of each variable's importance were made using the absolute values of coefficients derived from a multivariate linear regression model with extraction time as the outcome. Appropriate uni-, bi , and multivariate statistics were computed. RESULTS: The sample consisted of 14 surgeons who removed 450 M3s from 150 subjects from June 2002 to August 2003. Based on the multivariate linear regression model, variables associated with M3 extraction time were gender, arch location, Winter's classification, tooth morphology, number of teeth extracted, procedure type, and surgical experience. For these variables, there was a strong, statistically significant correlation (r = 0.86; P <.01) between the standardized coefficient absolute values and the surgeons' estimates of importance. CONCLUSION: There was a large, positive correlation between variables that surgeons consider most important in determining M3 extraction difficulty and those exhibiting the most influence over extraction times in a multivariate model. PMID- 15789313 TI - Glandular odontogenic cyst: treatment and recurrence. AB - PURPOSE: To investigate the correlation between clinical characteristics, radiologic features, treatment modalities, and treatment outcome of glandular odontogenic cyst, and to suggest a treatment protocol based on these results. PATIENTS AND METHODS: The study included a total of 56 cases, 49 from the literature and 7 new cases. Demographic data, locularity and radiographic extension, cortical plate integrity, treatment modalities, follow-up, and recurrence were analyzed. RESULTS: There were 34 male and 22 female patients aged 14 to 74 years (mean, 48 years). The mandible was involved in 41 cases (73.2%) and the maxilla in 15 (26.8%), predominantly in the anterior region; 53.6% of the lesions were unilocular and 46.4% multilocular. Large lesions were found in 78.5% of cases. Cortical integrity was compromised in 53.6% (cortical perforation in 39.3% and thinning or erosion of the cortical plate in 14.3%). Recurrence occurred at a rate of 29.2%, within 0.5 to 7 years (mean, 2.9 years). Mean follow up was also 2.9 years. Two patients had 3 recurrences each. Recurrence was associated with minor surgery such as enucleation or curettage; none of the patients treated by peripheral ostectomy, marginal resection, or partial jaw resection had a recurrence. Compared with the patients without recurrence, the recurrence group had a higher frequency of multilocularity than the nonrecurrent group (64.3% vs 41.2%) and of compromised cortical integrity (71.4% vs 47.1%). CONCLUSION: Glandular odontogenic cyst is an aggressive lesion. Treatment by enucleation or curettage alone is associated with a high recurrence rate. Small unilocular lesions can be treated by enucleation. In large uni- or multilocular lesions, an initial biopsy is recommended. Surgical treatment of large lesions should include enucleation with peripheral ostectomy for unilocular cases and marginal resection or partial jaw resection in multilocular cases. Marsupialization followed by second phase surgery is an option for lesions approaching vital structures. Follow-up should continue for at least 3 years (up to 7 years in cases with features associated with increased risk). PMID- 15789314 TI - Changes in the chewing path of patients in skeletal class III with and without asymmetry before and after orthognathic surgery. AB - PURPOSE: The purpose of this study was to examine the relation between changes in the condylar long axis and the chewing path before and after mandibular ramus osteotomy for patients with prognathism with and without asymmetry. PATIENTS AND METHODS: Eleven men and 16 women with mandibular prognathism were divided into groups on the basis of symmetry and osteotomy procedure. Preoperative and postoperative frontal chewing paths were recorded. The chewing path consisted of 4 components: deviated side range, undeviated side range, vertical range, and incisal path angle. The angle of the condylar long axis and the 4 components of the chewing path were compared between groups and the differences were analyzed statistically. RESULTS: No significant differences in each of the 4 chewing path components were found between groups on the basis of symmetry or osteotomy procedure. A positive correlation was found between the changes in incisal path angle when chewing on the undeviated side and condylar long axis angle on the undeviated side (P < .05). A positive correlation was also found between the changes in undeviated side range when chewing on the deviated side and condylar long axis angle on the deviated side (P < .05). CONCLUSION: This study suggests that surgically induced increase in the condylar long axis is correlated with increase in side range and incisor path angle, although surgical orthodontic treatment does not significantly change the chewing pattern. PMID- 15789315 TI - A qualitative report of patient problems and postoperative instructions. AB - PURPOSE: While surgery related stress may interfere with the patient's ability to concentrate on instructions, language difficulty or low health literacy may also impede appropriate doctor/patient communication. The purpose of this study is to understand from a sample of minority patients the types of problems encountered during healing and the level of information regarding elements of postoperative instructions they recalled receiving at an inner-city safety net hospital. We initiated a qualitative study to understand the care sequence process and provision of informed consent and postoperative instruction. METHODS: African American or Latino patients, 18 years of age or older, who had third molars removed under general anesthesia or received treatment for a mandibular fracture were recruited to participate in a focus group to discuss their treatment. Patients described their problem and any informed consent given about treatment risks and benefits and postoperative information they recalled. RESULTS: A total of 137 former patients were approached, 57 agreed to participate (42%) and 34 of those (60%) completed the interview. Subjects included 14 females and 20 males. Five categories of patient problems were reported: physical, eating, treatment related, psychosocial, and other problems. People reported 5 categories of coping strategies: medication use, physical treatments, dietary solutions, rest, and clinical assistance. Twenty people recalled being given informed consent, and 5 participants recalled no elements of informed consent. Overall, 14 participants recalled elements of postoperative instruction. CONCLUSION: Gaps in patient understanding of postoperative care suggest room for improvement in postoperative instructions. Additional research is necessary to design and test high-quality postoperative instructions for surgical treatment and recovery in populations with limited health related literacy. PMID- 15789316 TI - Evaluation of patient-controlled remifentanil application in third molar surgery. AB - PURPOSE: The aim of this study was to evaluate the clinical efficacy and safety of patient-controlled remifentanil application in combination with intravenous (IV) midazolam sedation during third molar surgery. PATIENTS AND METHODS: Twenty healthy patients with symmetrically placed impacted bilateral mandibular third molars were included in this prospective, placebo-controlled, double-blind, cross over, randomized clinical study. A bolus dose of 0.05 mg/kg intravenous midazolam was applied to each patient. Remifentanil was administered (group MR) by patient controlled infusion (PCI) either in the first or the second operation. In the other operation, a placebo (group MP) was given in the same manner. Perioperative blood samples were obtained to determine the changes in the stress hormone (aldosterone, adrenocorticotropic hormone [ACTH], renin) levels. Vital signs and oxygen saturation were recorded. Ramsey's sedation scale and modified Steward's recovery scale was used for evaluation of consciousness. Pain, patient satisfaction, cooperation score, reaction to local anesthetic injection, and degree of amnesia were also assessed. RESULTS: Remifentanil significantly increased the level of sedation, patient cooperation, and satisfaction; decreased the oxygen saturation, heart rate, and pain. While renin levels were significantly different, ACTH and aldosteron levels did not differ significantly between the 2 groups. Aldosterone and ACTH levels significantly decreased, whereas the renin level significantly increased in both groups during the operations. Remifentanil did not significantly alter the recovery time. CONCLUSION: Patient-controlled remifentanil application in combination with IV midazolam sedation seems to be a safe and reliable method, which effectively eliminates pain and provides a satisfactory sedation level, without any serious side effect. PMID- 15789317 TI - Minimally invasive tibial bone harvesting under intravenous sedation. AB - PURPOSE: Collecting high amounts of autogenous bone often results in considerable donor site morbidity. The hypothesis evaluated with this prospective study is that a modified approach for tibial bone harvesting using a minimally invasive access under local anesthesia plus sedation in an office setting compares favorably in terms of amount of bone harvested, morbidity, and patient satisfaction with more aggressive approaches previously reported. PATIENTS AND METHODS: Thirty-eight patients (18 women, 10 men) were treated using this method and followed prospectively. A medial approach to the proximal tibia was performed in all cases. A 10 mm incision gives access to an 8 mm manual trephine, which creates a bony window. Cancellous bone is released from the proximal compartment and a bone filter connected to suction allows fast removal of bone particles. Amount of bone harvested (compressed and non-compressed), surgical time, and complications were recorded. RESULTS: Mean surgical time was 14 minutes (range, 9 to 20 minutes). Volume of compressed cancellous bone ranged between 18 and 30 cc (mean, 28 cc). CONCLUSION: Tibial bone harvesting through a medial minimally invasive approach with a bone filter yields satisfactory results in terms of bone volume, surgical time, and patient satisfaction. PMID- 15789318 TI - Arthroscopic lysis and lavage in different stages of internal derangement of the temporomandibular joint: correlation of preoperative staging to arthroscopic findings and treatment outcome. AB - PURPOSE: The study was designed to evaluate the outcome of standard arthroscopic lysis and lavage for internal derangement with various levels of severity by comparing the preoperative staging with arthroscopic findings and subsequent success rates. PATIENTS AND METHODS: Temporomandibular joint disorder in 23 patients (26 joints) who underwent arthroscopic lysis and lavage was preoperatively classified as Wilkes stages II-V based on the clinical and radiologic (magnetic resonance imaging) findings. Recorded arthroscopic findings were scored and compared with the stages. The patients were examined both preoperatively and after a mean follow-up of 22.7 months, using objective and subjective criteria. RESULTS: The arthroscopic findings showed a correlation between increasing scores and advancing stage. Postoperatively, the patients could be clearly classified into 2 groups with either satisfactory or poor clinical outcome. Overall success rate was 78.3% (18/23). The success rates were slightly lower for patients with advanced stages than for those of stages II and III. Patients totally unresponsive to the treatment were found in all stages. CONCLUSION: Arthroscopic lysis and lavage is a preferred treatment for different stages of internal derangement. Preoperative staging and corresponding characteristics of the arthroscopic findings do not seem to correlate with the prognosis of the treatment outcome. PMID- 15789319 TI - Positron emission tomography/computed tomography true fusion imaging in clinical head and neck oncology: early experience. AB - PURPOSE: Positron emission tomography-computed tomography (PET-CT) is a new imaging modality that provides simultaneous functional and anatomic information. Its application in head and neck oncology is still evolving. MATERIALS AND METHODS: Five illustrative cases who were scanned with this technique are presented. RESULTS: The clinical application and impact on clinical management are described and discussed. CONCLUSION: Our experience indicates that PET-CT will assume a more important role than either CT, magnetic resonance imaging, or PET alone in head and neck cancer imaging. PMID- 15789320 TI - Comparison of 2 preferred methods used for frontal sinus obliteration. AB - PURPOSE: This study was undertaken to compare total operating room cost, total operating time, and potential complications in frontal sinus obliteration using 2 different techniques. PATIENTS AND METHODS: Hospital records of all patients with frontal sinus fractures treated by the Division of Maxillofacial Surgery at the University of Florida, Jacksonville between October 1998 and December 2003 were reviewed. Twelve patients required frontal sinus obliteration based on the severity and location of injury. All cases were caused by traumatic etiology. Patients were divided into group A or B. Six patients (group A) were treated using autogenous abdominal fat for obliteration purposes, while the other 6 patients (group B) underwent frontal sinus obliteration using a hydroxyapatite cement. Total operating cost, total operating time, and any complications were recorded and analyzed for each group and then statistically evaluated using a t test. RESULTS: Follow-up ranged from 2 weeks to 6 months. Patients in group A had a lower total operating cost compared with group B. This cost difference was statistically significant. Total operating time was slightly greater in group A versus group B, although this was not statistically significant. CONCLUSION: Frontal sinus obliteration using autogenous abdominal fat appears to be more cost effective compared with hydroxyapatite cement. The slight difference in total operating time was not statistically significant and this factor alone should not be a deterrent from performing this surgical procedure. PMID- 15789321 TI - A novel high-resolution magnetic resonance imaging microscopy coil as an alternative to the multislice computed tomography in postoperative imaging of orbital fractures and computer-based volume measurement. AB - BACKGROUND: Multislice computed tomography (MSCT) has been the modality of choice for postoperative detailed imaging of orbital trauma. Unfortunately, it involves extensive exposition of the lens to radiation, especially when taking multiple readings. Also, it holds beam hardening effects and limited imaging (delineation) of the reconstruction material. Alternative conventional magnetic resonance imaging (MRI) head coils (MRIhc) present reduced differentiation of anatomic structures caused by low signal and artifact appearance. A substantially improved depiction is made possible by a newly introduced MRI microscopy coil (MRImc), used for the first time in this field. PATIENTS AND METHODS: In this prospective study, 32 patients with extended orbital wall fractures (n = 36) were treated surgically using a polydioxanonsulfate (PDS) foil after reconstruction. Postoperatively, imaging was performed using MRImc, conventional MRI, and MSCT to evaluate the different imaging techniques. RESULTS: The position of the PDS foil could precisely be depicted in 29 of 36 fractures by MRImc, whereas by conventional MRI and MSCT the reconstruction material could only be detected in 25 and 24 of 36 fractures, respectively. In contrast to MRIhc, the new microscopy coil allows fast and high resolution imaging and therefore a clear differentiation of eventual postoperative complications (eg, dislocation of the PDS foil with secondary soft tissue entrapment resulting in limitation of ocular movements or enophthalmos). In 13 long lasting symptomatic cases, revision surgery could be avoided because of regular MRImc findings, apart from muscle swelling and hematoma. In 11 cases of inadequate PDS foil position resulting in revision, the mean volume of displaced tissue (VDT) of 0.62 cm3 correlated significantly to an enophthalmos of more than 2 mm. CONCLUSION: In this pilot study, MRImc proved to be highly superior to MRIhc and MSCT in postsurgical orbital imaging, especially for decision making regarding revision surgery. PMID- 15789322 TI - Tensile mechanical strain up-regulates Runx2 and osteogenic factor expression in human periosteal cells: implications for distraction osteogenesis. AB - PURPOSE: Distraction osteogenesis is now accepted as a standard treatment in oral and maxillofacial reconstructive surgery. In the process of bone regeneration with the application of strain, the periosteum might be very involved in osteogenesis. This study examined the effect of mechanical strain on periosteal cells and the implications for distraction osteogenesis. MATERIALS AND METHODS: Periosteal cells were obtained from mandibular periosteum that was excised while extracting impacted wisdom teeth. Mechanical strain was applied using a specially designed apparatus with flexible silicon bottom chambers. The levels of mRNA of the osteoblast differentiation factor Runx2 (Cbfa1/AML3/Peb alpha A) and osteogenic factors were analyzed at different times using the reverse transcription-polymerase chain reaction method to evaluate the effect of the strain. RESULTS: The periosteal cells expressed the osteogenic phenotype. The strain had a shaping effect on the cells. The application of tensile strain strongly activated the expression of osteogenic and angiogenic growth factors, and up-regulated the expression of Runx2, an osteoblast-specific transcription factor. CONCLUSION: Tensile strain may initiate the differentiation of periosteal cells into osteogenic cells, inducing the expression of Runx2 in the process of bone regeneration. Therefore, the periosteum is profoundly involved in bone formation and regeneration, especially in distraction osteogenesis. PMID- 15789323 TI - Changes in parathyroid hormone-related protein and 3-dimensional trabecular bone structure of the mandibular condyle following mandibular distraction osteogenesis in growing rats. AB - PURPOSE: Distraction osteogenesis (DO) is commonly performed for mandibular reconstruction during the growth period. We tested the hypothesis that parathyroid hormone-related protein (PTHrP) in mandibular condylar cartilage and underlying trabecular bone in growing individuals undergo changes in response to distraction forces. MATERIALS AND METHODS: Forty-eight 6-week-old male Sprague Dawley rats were used. Animals underwent unilateral mandibular distraction using a distractor that we devised, and unoperated animals were evaluated as controls. DO procedure was performed: 3 days' latency period, 0.4 mm/day rate, total 4.0 mm. Changes in cartilage morphology, PTHrP activity, and 3-dimensional trabecular bone structure changes measured by micro-computed tomography were examined at 0, 2, 4, and 6 weeks of consolidation. RESULTS: A marked irregularity was noted in the superior portion of the distracted side's condylar cartilage that resolved after distraction ceased. PTHrP was more strongly expressed in the hypertrophic layer of condylar cartilage on the distracted side than in controls, up to 6 weeks after the end of distraction. Subchondral trabecular bone volume, percent bone volume, and trabecular number in the superior and posterior regions of the condyle decreased significantly by 2 weeks after distraction. These parameters returned to normal in the posterior condyle, but not in the superior part of the condyle by 6 weeks following distraction. CONCLUSION: These results suggest that unilateral mandibular distraction in growing rats causes temporary morphologic alterations of trabecular bone structure on the distracted side accompanied by increased production of PTHrP in the mandibular condyle. PMID- 15789324 TI - Sociodemographic risk indicators for depressive symptoms among persons with oral cancer or oral epithelial dysplasia. AB - PURPOSE: We report findings from a study that measured associations between sociodemographic risk indicators and depressive symptoms among individuals diagnosed with either oral cancer or a premalignant lesion. MATERIALS AND METHODS: Incident cases of oral cancer and oral epithelial dysplasia (OED) were identified by reviewing pathology reports generated by 3 oral pathology laboratories serving primarily community-based oral and maxillofacial surgeons. Subjects were interviewed by telephone to collect information on sociodemographic characteristics, depressive symptoms using the Center for Epidemiologic Studies Depression (CES-D) Scale, and social support using the Berkman Social Network Inventory. RESULTS: The analysis included 167 oral cancer and 234 OED cases. Nineteen percent of the subjects had a CES-D score indicative of clinical depression (CES-D > or =16). Forward and backward stepwise logistic regression identified diagnosis (cancer/OED), age, social support, employment status, and gender as sociodemographic indicators of CES-D scores of 16+. In the final model, which also controlled for smoking and drinking, the odds of having elevated CES-D scores (16+) were 79% higher among oral cancer relative to OED cases. The odds of high CES-D scores were significantly reduced in persons over the age of 50 compared with those aged 50 years and younger as well as in persons with higher, relative to low, levels of social support and in persons employed outside the home compared with those who were not. Although not statistically significant, men were more likely to have CES-D scores indicative of clinical depression. CONCLUSIONS: Knowledge of sociodemographic characteristics may assist the clinician in identifying those individuals with an elevated risk of concomitant depressive symptoms. PMID- 15789325 TI - Differential growth factor retention by platelet-rich plasma composites. AB - PURPOSE: This study evaluates the temporal sequence and growth factor release from platelet-rich plasma (PRP) combined with different bone substitutes (BS), to identify an optimal substrate for extended growth factor retention. MATERIALS AND METHODS: PRP was clotted with bovine thrombin or thrombin receptor activator peptide-6 (TRAP). In addition, PRP was clotted using Allogro (Ceramed, Lakewood, CO), BioGlass (Mo-Sci, Rolla, MN), or BioOss (Osteohealth, Shirley, NY). The effects of media exchange and BS on platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF beta) release were quantified via enzyme linked immunosorbent assay. RESULTS: At day 1, the thrombin group released 36% more PDGF than the TRAP group and 80% more than the BS groups. At 7 days, PDGF release was the greatest for the TRAP group. PDGF release was minimal for all groups at day 14, with BS groups retaining 60% more PDGF than thrombin clots. Similarly, the thrombin group released the greatest amount of TGF beta (81.4% of the total), whereas TRAP and BS groups released significantly less TGF beta at day 1. Compared with thrombin, TRAP retained 39.2% more TGF beta, whereas BS groups retained even greater levels (Allogro, 54.3%; BioOss, 45.8%; BioGlass, 67.0%). No significant difference in TGF beta release was observed among the substitutes after day 1. The BS groups continued to retain TGF beta after 14 days, whereas all TGF beta in the thrombin clots was depleted. CONCLUSIONS: PRP preparation with thrombin results in a large, immediate release of growth factors that could be lost into the interstitium in vivo. TRAP-BS may prove more efficacious than thrombin in sustaining growth factor levels critical for the cascade of events leading to bone formation. PMID- 15789326 TI - Activation of platelet-rich plasma using thrombin receptor agonist peptide. AB - PURPOSE: This study proposes an alternative preparation method of platelet-rich plasma (PRP). Specifically, we compare the use of thrombin receptor agonist peptide-6 (TRAP) and bovine thrombin as a clotting agent in the preparation of PRP. MATERIALS AND METHODS: PRP was prepared by centrifugation and clotted with thrombin or TRAP. In vitro clotting times were monitored as a function of TRAP concentration, and clot retraction was determined by measuring clot diameter over time. Following the optimization of TRAP concentration, experiments were repeated with the addition of several commercially available bone substitutes. The release of PRP-relevant growth factors as a function of PRP preparation was also determined. RESULTS: The most rapid polymerization of PRP takes place with the addition of thrombin, followed by TRAP/Allogro (Ceramed, Lakewood, CO), TRAP/BioGlass (Mo-Sci, Rolla, MN), TRAP/BioOss (Osteohealth, Shirley, NY), and TRAP alone. Thrombin caused considerable clot retraction (43%), whereas TRAP alone resulted in only 15% retraction. TRAP/Allogro, TRAP/BioOss, and TRAP/BioGlass all exhibited minimal retraction (8%). CONCLUSIONS: The use of TRAP to activate clot formation in the preparation of PRP may be a safe alternative to bovine thrombin. It results in an excellent working time and significantly less clot retraction than the currently available methods of PRP production. PMID- 15789327 TI - Evidence to support an individualized approach to modification of oral anticoagulant therapy for ambulatory oral surgery. PMID- 15789328 TI - Evidence to continue oral anticoagulant therapy for ambulatory oral surgery. PMID- 15789329 TI - Malignant fibrous histiocytoma of the tongue: review of the literature and report of a case. PMID- 15789330 TI - Intraoperative angioedema induced by angiotensin-converting enzyme inhibitors: overview and case report. PMID- 15789331 TI - Unusual presentation of a cervical pseudomeningocele: a case report and review of the literature. PMID- 15789332 TI - Bilateral parotid diffuse hyperplastic oncocytosis: case report. PMID- 15789333 TI - A straight-line incision for neck dissection: technical note. PMID- 15789334 TI - Extended bilaminar forehead flap with cantilevered bone grafts for reconstruction of full-thickness nasal defects. PMID- 15789335 TI - Educational purpose and content: more on the offshore medical degree. PMID- 15789336 TI - Antibiotic usage for coronectomy: is it necessary? PMID- 15789338 TI - Polymerase mu is up-regulated during the T cell-dependent immune response and its deficiency alters developmental dynamics of spleen centroblasts. AB - Mammalian DNA polymerase mu (Polmu), preferentially expressed in secondary lymphoid organs, is shown here to be up-regulated in germinal centers after immunization. Alternative splicing appears to be part of Polmu regulation during an immune response. We generated Polmu-deficient mice that are viable and show no anatomical malformation or serious alteration in lymphoid populations, with the exception of an underrepresentation of the B cell compartment. Young and aged homozygous Polmu(-/-) mice generated similar immune responses after immunization with the hapten (4-hydroxy-3-nitrophenyl)acetyl (NP) coupled to chicken gammaglobulin (CGG), compared with their wild-type littermates. Nonetheless, the kinetics of development of the centroblast population showed significant differences. Hypermutation analysis of the rearranged heavy chain intron region in centroblasts isolated from NP-CGG-immunized Polmu(-/-) mice showed a similar quantitative and qualitative somatic mutation spectrum, but a lower representation of heavily mutated clones. These results suggest that although it is not a critical partner, Polmu modulates the in vivo somatic hypermutation process. PMID- 15789337 TI - p53 is required for spontaneous autoantibody production in B6/lpr lupus mice. AB - The p53 tumor suppressor molecule triggers a key pathway of apoptosis in injured cells, in part through induction of Fas. The importance of Fas as a receptor mediating apoptosis is highlighted by the lupus-like systemic autoimmunity seen in animals and humans with nonfunctional Fas molecules. We set out to see if the absence of p53, superimposed on the Fas defect of lpr mice, might further accelerate or exacerbate their systemic autoimmunity. We generated double mutant mice (p53(-/-) lpr) having defects in both p53- and Fas-dependent pathways, hypothesizing that animals with lesions in both Fas- and p53-dependent pathways would show reduced ability to delete autoreactive or injured cells, thereby producing more severe autoimmune disease. Surprisingly, these mice have lower autoantibody levels than the single mutant lpr mice. These studies suggest an unanticipated role for p53 in the progression of autoimmunity and the production of autoantibodies. PMID- 15789339 TI - Antitumor immune response by CX3CL1 fractalkine gene transfer depends on both NK and T cells. AB - The CX3C chemokine fractalkine (CX3CL1) exists as both a membrane-bound form promoting firm cell-cell adhesion and a soluble form chemoattracting leukocytes expressing its receptor CX3CR1. When adenoviral vector expressing mouse fractalkine (AdFKN) was transduced to the tumor cells, fractalkine was expressed as both membrane-bound form on the tumor cells and soluble form in the supernatant in vitro. Intratumoral injection of AdFKN (1 x 10(9)PFU/tumor) into C26 and B16F10 tumors resulted in marked reduction of tumor growth compared to control (C26: 86.5%, p<0.001; B16F10: 85.5%, p<0.001). Histological examination of tumor tissues revealed abundant infiltration of NK cells, dendritic cells, and CD8(+) T lymphocytes 3 and/or 6 days after treatment with AdFKN. Splenocytes from mice treated by AdFKN developed tumor-specific cytotoxic T cells, and thereby protected from rechallenging with parental tumor cells. Antitumor effects by AdFKN were completely abrogated in both NK cell-depleted mice and CD8(-/-) mice, and partially blocked in CD4(-/-) mice. These data indicated that fractalkine mediates antitumor effects by both NK cell-dependent and T cell-dependent mechanisms. This study suggests that fractalkine can be a suitable candidate for immunogene therapy of cancer because fractalkine induces both innate and adaptive immunity. PMID- 15789340 TI - Increased inflammation in mice deficient for the chemokine decoy receptor D6. AB - Chemokines are chemotactic cytokines with a key role in the control of cell trafficking and positioning under homeostatic and inflammatory conditions. D6 is a promiscuous 7-transmembrane-domain receptor expressed on lymphatic vessels which recognizes most inflammatory, but not homeostatic, CC chemokines. In vitro experiments demonstrated that D6 is unable to signal after ligand engagement, and it is structurally adapted to sustain rapid and efficient ligand internalization and degradation. These unique functional properties lead to the hypothesis that D6 may be involved in the control of inflammation by acting as a decoy and scavenger receptor for inflammatory chemokines. Consistent with this hypothesis, here we report that D6(-/-) mice showed an anticipated and exacerbated inflammatory response in a model of skin inflammation. Moreover, the absence of D6 resulted in increase cellularity and inflammatory-chemokine levels in draining lymph nodes. Thus, D6 is a decoy receptor structurally adapted and strategically located to tune tissue inflammation and control transfer of inflammatory chemokines to draining lymph nodes. PMID- 15789341 TI - Overexpression of cathepsin B in gastric cancer identified by proteome analysis. AB - We aimed to validate an analytical approach based on proteomics on gastric cancer specimens for the identification of new putative diagnostic or prognostic markers. Primary screening was performed on gastrectomy specimens obtained from ten consecutive patients with gastric cancer. Gastric epithelial cells were obtained with an epithelial cell enrichment technique, homogenized and then separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The differential protein expression pattern was verified stepwise by Western blotting and immunohistochemistry on samples from 28 and 46 cancer patients, respectively. The putative clinical applicability and prognostic use were tested by an enzyme linked immunoabsorbent assay on serum samples obtained from 149 cancer patients. One hundred-ninety-one differentially expressed protein spots were found by 2-D PAGE and identified by mass spectrometry, including cathepsin B, which was over expressed in six (60%) patients. Western blotting confirmed that the active form of cathepsin B is over-expressed, while immunohistochemistry showed strong cytoplasmic staining in cancer tissues of 45 (98%) patients. The serum level of cathepsin B was increased in patients with gastric cancer compared to healthy controls (P = 0.0026) and correlated with T-category and the presence of distant metastases (P < 0.05). Serum levels above 129 pmol x L(-1) were associated with a reduced survival rate (P = 0.0297). Proteome analysis is a valuable tool for the identification of prognostic markers in gastric cancer: Increased cathepsin B serum levels are associated with advanced tumor stages and progressive disease, which enables the classification of some gastric cancer patients into a subgroup that should undergo aggressive therapy. PMID- 15789342 TI - The identification of foam-forming soluble proteins from wheat (Triticum aestivum) dough. AB - Proteomic methods have been used to identify foam-forming soluble proteins from dough that may play an important role in stabilising gas bubbles in dough, and hence influence the crumb structure of bread. Proteins from a soluble fraction of dough (dough liquor) or dough liquor foam have been separated by two-dimensional gel electrophoresis, and 42 identified using a combination of matrix-assisted laser desorption/ionization-time of flight and quadrupole-time of flight analyses. Major polypeptide components included beta-amylase, tritin and serpins, with members of the alpha-amylase/trypsin inhibitor family being particularly abundant. Neither prolamin seed storage proteins nor the surface-active protein puroindoline were found. Commonly used dough ingredients (NaCl, Na L-ascorbate) had only a minor effect on the 2-DE protein profiles of dough liquor, of which one of the more significant was the loss of 9 kDa nonspecific lipid transfer protein. Many proteins were lost in dough liquor foam, particularly tritin, whilst a number of alpha-amylase inhibitors were more dominant, suggesting that these are amongst the most strongly surface-active proteins in dough liquor. Such proteins may play a role determining the ability of the aqueous phase of doughs, as represented by dough liquor, to form an elastic interface lining the bubbles, and hence maintain their integrity during dough proving. PMID- 15789343 TI - Proteomic analysis of reaper 5' untranslated region-interacting factors isolated by tobramycin affinity-selection reveals a role for La antigen in reaper mRNA translation. AB - Translational control is a key step in gene expression regulation during apoptosis. To understand the mechanisms of mRNA translation of a pro-apoptotic gene, reaper (rpr), we adapted the tobramycin-aptamer technique described by Hartmuth et al. (Proc. Natl. Acad. Sci. USA 2002, 99, 16719-16724) for the analysis of proteins interacting with rpr 5' untranslated region (UTR). We assembled ribonucleoprotein complexes in vitro using translation extracts derived from Drosophila embryos and purified the RNA-protein complexes for mas spectrometry analysis. We identified the proteins bound to the 5' UTR of rpr. One of them, the La antigen, was validated by RNA-crosslinking experiments using recombinant protein and by the translation efficiency of reporter mRNAs in Drosophila cells after RNAinterference experiments. Our data provide evidence of the involvement of La antigen in the translation of rpr and set a protocol for purification of tagged-RNA-protein complexes from cytoplasmic extracts. PMID- 15789344 TI - Differential display of proteins involved in the neural differentiation of mouse embryonic carcinoma P19 cells by comparative proteomic analysis. AB - Mouse embryonic carcinoma P19 cell has been used extensively as a model to study molecular mechanisms of neural differentiation in vitro. After retinoic acid (RA) treatment and aggregation, P19 cells can differentiate into neural cells including neurons and glial cells. In this study, comparative proteomic analysis is utilized to approach the protein profiles associated with the RA-induced neural differentiation of P19 cells. Image analysis of silver stained two dimensional gels indicated that 28 protein spots had significantly differential expression patterns in both quantity and quality. With mass spectrometry analysis and protein functional exploration, many proteins demonstrated an association with distinct aspects of neural differentiation. These proteins were gag polyprotein, rod cGMP-specific 3',5'-cyclic phosphodiesterase, 53 kDa BRG1 associated factor A, N-myc downstream regulated 1, Vitamin D receptor associated factor 1, stromal cell derived factor receptor 1, phosphoglycerate mutase, Ran specific GTPase-activating protein, and retinoic acid (RA)-binding protein. While some cytoskeleton-related proteins such as beta cytoskeletal actin, gamma-actin, actin-related protein 1, tropomyosin 1, and cofilin 1 are related to cell migration and aggregation, other proteins have shown a relationship with distinct aspects of neural differentiation including energy production and utilization, protein synthesis and folding, cell signaling transduction, and self-protection. The differential expression patterns of these 28 proteins indicate their different roles during the neural differentiation of P19 cells. As an initial step toward unveiling the regulations involved in the commitment of pluripotent cells to a neural fate, information from this study may be helpful to uncover the molecular mechanisms of neural differentiation. PMID- 15789345 TI - The human brain mannose 6-phosphate glycoproteome: a complex mixture composed of multiple isoforms of many soluble lysosomal proteins. AB - The lysosome is a membrane delimited cytoplasmic organelle that contains at least 50 hydrolytic enzymes and associated cofactors. The biomedical importance of these enzymes is highlighted by the many lysosomal storage disorders that are associated with mutations in genes encoding lysosomal proteins, and there is also evidence that lysosomal activities may be involved in more widespread human diseases. The aim of this study was to characterize the human brain lysosomal proteome with the goal of establishing a reference map to investigate human diseases of unknown etiology and to gain insights into the cellular function of the lysosome. Proteins containing mannose 6-phosphate (Man6-P), a carbohydrate modification used for targeting resident soluble lysosomal proteins to the lysosome, were affinity-purified using immobilized Man6-P receptor. Fractionation by two-dimensional electrophoresis resolved a complex mixture comprising approximately 800 spots. Constituent proteins in each spot were identified using a combination of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (both peptide mass fingerprinting and tandem mass spectrometry) [corrected] on in-gel tryptic digests and N-terminal sequencing. In a complementary analysis, we also analyzed a tryptic digest of the unfractionated mixture by liquid chromatography MS/MS. In total, 61 different proteins were identified. Seven were likely contaminants associated with true Man6-P glycoproteins. Forty-one were known lysosomal proteins of which 11 have not previously been reported to contain Man6-P. An additional nine proteins were either uncharacterized or proteins not previously reported to have lysosomal function. We found that the human brain Man6-P-containing lysosomal proteome is highly complex and contains more proteins with a much greater number of individual isoforms than found in previous studies of Man6-P glycoproteomes. PMID- 15789346 TI - Post-translational processing of beta-secretase in Alzheimer's disease. AB - Beta-amyloid is released into the brains of Alzheimer's patients, where it aggregates and causes damage to neurons. It is cleaved proteolytically from a large transmembrane glycoprotein amyloid precursor protein by a membrane-bound protease, known as beta-secretase identified previously as the acid protease, Asp 2. We have shown previously that beta-secretase is up-regulated by increased intracellular cholesterol, and down-regulated by cholesterol biosynthesis inhibition. Here we show using mass spectrometry that discrete changes in the glycosylation and palmitoylation of beta-secretase occur when cells expressing it are treated with statins. PMID- 15789347 TI - Stress-induced changes in the Schizosaccharomyces pombe proteome using two dimensional difference gel electrophoresis, mass spectrometry and a novel integrated robotics platform. AB - Robotic and manual methods have been used to obtain identification of significantly changing proteins regulated when Schizosaccharomyces pombe is exposed to oxidative stress. Differently treated S. pombe cells were lysed, labelled with CyDye and analysed by two-dimensional difference gel electrophoresis. Gel images analysed off-line, using the DeCyder image analysis software [GE Healthcare, Amersham, UK] allowed selection of significantly regulated proteins. Proteins displaying differential expression were excised robotically for manual digestion and identified by matrix-assisted laser desorption/ionisation - mass spectrometry (MALDI-MS). Additionally the same set of proteins displaying differential expression were automatically cut and digested using a prototype robotic platform. Automated MALDI-MS, peak label assignment and database searching were utilised to identify as many proteins as possible. The results achieved by the robotic system were compared to manual methods. The identification of all significantly altered proteins provides an annotated peroxide stress-related proteome that can be used as a base resource against which other stress-induced proteomic changes can be compared. PMID- 15789348 TI - Cloning and disruption of the Pichia pastoris ARG1, ARG2, ARG3, HIS1, HIS2, HIS5, HIS6 genes and their use as auxotrophic markers. AB - Screening of a partial genomic database of Pichia pastoris allowed us to identify the ARG1, ARG2, ARG3, HIS1, HIS2, HIS5 and HIS6 genes, based on homology to their Saccharomyces cerevisiae counterparts. Based on the cloned sequences, a set of disruption vectors was constructed, using the previously described PpURA5-blaster as a selectable marker, and the cloned genes were individually disrupted. All disruptants exhibited the expected auxotrophic phenotypes, with only the his2 knockouts displaying a bradytroph phenotype. To allow their use as auxotrophic markers, we amplified the open reading frames and respective promoters and terminator regions of PpARG1, PpARG2, PpARG3, PpHIS1, PpHIS2 and PpHIS5. We then designed a set of integration vectors harbouring cassettes of the ARG pathway as selectable markers, to disrupt the genes of the HIS pathway and vice versa. Employing this strategy, we devised a scheme allowing for the rapid and stable introduction of several heterologous genes into the genome of P. pastoris without the need for recyclable markers or strains with multiple auxotrophies. Furthermore, simple replica-plating, instead of cost-consuming and labour intensive colony PCR or Southern analysis, can be used to identify positive transformants, making this approach amendable for initial high-throughput applications, which can then be followed up by a more careful analysis of the selected transformants. PMID- 15789349 TI - Deletion of PDE2, the gene encoding the high-affinity cAMP phosphodiesterase, results in changes of the cell wall and membrane in Candida albicans. AB - A role for the cAMP-dependent pathway in regulation of the cell wall in the model yeast Saccharomyces cerevisiae has recently been demonstrated. In this study we report the results of a phenotypic analysis of a Candida albicans mutant, characterized by a constitutive activation of the cAMP pathway due to deletion of PDE2, the gene encoding the high cAMP-affinity phosphodiesterase. Unlike wild type strains, this mutant has an increased sensitivity to cell wall and membrane perturbing agents such as SDS and CFW, and antifungals such as amphotericin B and flucytosine. Moreover, the mutant is characterized by an altered sensitivity and a significantly reduced tolerance to fluconazole. The mutant's membrane has around 30% higher ergosterol content and the cell wall glucan was 22% lower than in the wild-type. These cell wall and membrane changes are manifested by a considerable reduction in the thickness of the cell wall, which in the mutant is on average 60-65 nm, compared to 80-85 nm in the wild-type strains as revealed by electron microscopy. These results suggest that constitutive activation of the cAMP pathway affects cell wall and membrane structure, and biosynthesis, not only in the model yeast S. cerevisiae but also in the human fungal pathogen C. albicans. PMID- 15789350 TI - Inverted repeat of a large segment unveiled on the right arm of Saccharomyces cerevisiae chromosome II. AB - By means of gene disruption analyses, Saccharomyces cerevisiae strain YPH499 was shown to have two, and only two, copies of ATP3 that encodes the gamma-subunit of H+-dependent ATP synthase and locates on the right arm of chromosome II. Linkage analyses of the two distinguishably marked copies of ATP3 indicated that the distance between them was about 43 cM. Since YBR030W, an ORF proximal to ATP3 by a distance of 17 kbp, was also found to be duplicated, we marked them with two distinguishable nutritional markers, which were also distinguishable from those used for marking the two copies of ATP3, and achieved four-point linkage analyses; CEN2 marked with an appropriate nutritional marker gene was included as a reference point. And, the following linkage map was deduced: CEN2- [11 cM] YBR030Wa- [8 cM]-ATP3a-[47 cM]-ATP3b- [55 cM]-YBR030Wb. From this map, we suspected that a segment spanning at least YBR030W-ATP3 would be inversely duplicated on the right arm of chromosome II. We then carried out chromosome fragmentation analyses, using several laboratory strains including YPH499, and obtained data in accord with our speculation for all strains, although the distance between the two copies of ATP3 varied from 48 kbp to 192 kbp among the strains examined. PMID- 15789351 TI - Expression of GCR1, the transcriptional activator of glycolytic enzyme genes in the yeast Saccharomyces cerevisiae, is positively autoregulated by Gcr1p. AB - When regulation of GCR1 expression was analysed using a GCR1-lacZ fusion, lacZ expression levels were decreased in the Deltagcr1 or Deltagcr2 mutant. RT-PCR analysis of genomic GCR1 transcript confirmed the dependency of GCR1 expression on the Gcr1p-Gcr2p complex. Examination of the 5' non-coding region of GCR1 identified three putative Gcr1p binding sites (CT-boxes) in the -100 to -200 region of GCR1, and the putative binding sites for Rap1p (RPG-box) and Abf1p were also identified nearby. The region containing putative cis-elements was analysed by cloning it upstream of the CYC1TATA-lacZ fusion. The GCR1(UAS)-CYC1TATA-lacZ fusion showed a moderate activity and, as expected, the activity was drastically reduced in the Deltagcr1 or Deltagcr2 mutant. Systematic deletion and mutation analyses of cis-elements in this region demonstrated that the putative binding sites for Rap1p and Abf1p were not involved in the promoter activity of GCR1(UAS) and only one of the three CT-boxes showed GCR1- and GCR2-dependent promoter activity. In contrast to the expression of glycolytic genes, where a RPG-box adjacent to the CT-box is required for strong promoter activities, CT-box dependent expression of GCR1 did not require the RPG-box. Also, a contribution of Sgc1p, an E-box binding transcription factor, to the expression of GCR1 was suggested, based on its disruption analysis. PMID- 15789352 TI - Ultrasound-assisted laparoscopic cryomyolysis: two- and three-dimensional findings before, during and after treatment. AB - OBJECTIVE: To investigate the role of two- and three-dimensional (2D and 3D) ultrasound and power Doppler before, during and after surgery in monitoring the effects of uterine fibroid laparoscopic cryomyolysis. METHODS: This prospective study involved 10 premenopausal patients with a sonographic diagnosis of a single subserosal and/or intramural uterine myoma, who underwent laparoscopic cryomyolysis. All patients suffered from symptoms due to fibroids (menometrorrhagia, bulk-related symptoms, pelvic pain). During laparoscopy transvaginal sonography was performed to guide the insertion of the cryoprobe, monitor extension of the ice-ball and evaluate the reduction of the blood supply of the myoma. All patients underwent 2D and 3D sonographic and power Doppler imaging evaluation of the myoma 1 week before treatment, during cryomyolysis, and 1, 3 and 6 months after treatment; size, echostructure and vascularization were recorded. RESULTS: With cryomyolysis, we achieved selective vessel and tissue damage within the fibroid alone. Eight patients were free of symptoms and two had improved after 3 months. Progressive shrinkage of the treated myoma was observed during follow-up with a reduction percentage after 1 month of 22.2%, after 3 months of 37.5% and after 6 months of 52.6%. After cryomyolysis a significant reduction in central blood flow of the myoma was observed. 2D and 3D power Doppler evaluation of vascularization did not differ although subjectively findings were best evaluated by 3D images. CONCLUSION: Sonography can aid the safe accomplishment of cryomyolysis by assessing myomata preoperatively, guiding the freezing procedure during laparoscopy, and helping to monitor postoperative progress. The use of ultrasound in this new treatment of fibroids will permit the physician to modulate and individualize treatment. PMID- 15789353 TI - Middle cerebral artery peak systolic velocity for the diagnosis of fetal anemia: the untold story. PMID- 15789354 TI - Two- and three-dimensional sonographic diagnosis of incomplete uterine scar rupture during pregnancy. PMID- 15789355 TI - Extracellular lysosome-associated membrane protein-1 (LAMP-1) mediates autoimmune disease progression in the NOD model of type 1 diabetes. AB - Treatment (from 5 to 25 weeks of age) with a novel blocking monoclonal antibody, mAb I-10, directed against the plasma membrane (pm) form of LAMP-1, protected against development of autoimmune diabetes in the NOD mouse. A shorter course of treatment, i.e. from 5 to 12 weeks of age, significantly reduced the occurrence of insulitis as well as disease onset. Interfering with pm-LAMP-1 required continuous treatment as tolerance was not observed when treatment was stopped, and no higher proportion of cells with a T regulatory phenotype (e.g. CD4(+)CD25(+)) were induced. The mechanism appears to involve modulating a proinflammatory cytokine, as the proportion of pancreatic-infiltrating IFN-gamma positive cells was significantly reduced in the mAb I-10-treated group. These results demonstrate an unexpected role for pm-LAMP-1 in autoimmune disease progression, and suggest that further investigation should be performed to understand how this molecule modulates IFN-gamma-driven responses. PMID- 15789356 TI - Discovery and characterization of highly immunogenic and broadly recognized mimics of the HIV-1 CTL epitope Gag77-85. AB - Human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTL) play an important role in HIV infection. Given the viral genetic diversity, the selection of suitable antigens and epitope variants will be important in the design of an effective vaccine. We have previously shown that combinatorial libraries are useful tools to identify epitope mimics as well as potentially cross-reactive natural sequences in protein databases. We have applied this approach to the HIV Gag p17-derived epitope SL9 (SLYNTVATL) to identify broadly recognized SL9 mimics and to assess the cross-recognition of naturally occurring SL9 variants. Nine nonapeptides were identified that were up to one order of magnitude more effective than SL9 in stimulating CTL responses in PBMC from HIV-infected subjects. Using transgenic mice, we demonstrate that a number of these epitope mimics were able to generate de novo T cell responses that cross-reacted with the original SL9 sequence. Particularly, mimics with changes at the relatively conserved F-pocket anchor residue were frequently cross-recognized. This approach may lead to vaccine candidates with higher in vivo immunogenicity and increased potential for cross-recognition of naturally occurring SL9 variants. PMID- 15789357 TI - Prior elevation of IL-18 promotes rapid early IFN-gamma production during staphylococcal infection. AB - Systemic Staphylococcus aureus infection is associated with significant morbidity and mortality arising from both bacterial and host immune factors. IL-18 is a pro inflammatory cytokine of the IL-1 superfamily that exhibits broad functional effects in innate and acquired immune responses and which has been found in high levels in several chronic inflammatory and autoimmune diseases. Over-expression of IL-18 may promote early resolution of infection or could promote a detrimental exaggerated immune response. This was explored in a model of S. aureus infection. We report increased mortality in Swiss mice that were given recombinant IL-18 prior to inoculation with S. aureus LS-1. IL-18 administration prior to infection induced preferentially enhanced IFN-gamma mRNA expression in peripheral blood leukocytes and spleen, especially splenic NK cells. This correlated with increased IFN-gamma protein detection in serum, and leukocyte and spleen cultures at subsequent discrete time points. These data suggest that increased mortality following gram-positive infection in autoimmune diseases could in part reflect the impact of high levels of pleiotropic pro-inflammatory cytokines such as IL-18 present prior to the onset of infection. PMID- 15789358 TI - The binding affinity of double-stranded RNA motifs to HIV-1 Tat protein affects transactivation and the neutralizing capacity of anti-Tat antibodies elicited after intranasal immunization. AB - In this study we examined the hypothesis that the binding affinity of two double stranded (ds) RNA motifs to HIV-1 Tat protein might affect transactivation and the type of anti-Tat immune responses. Using surface plasmon resonance technology we demonstrated the capacity of the poly(A):poly(U) (pA:pU) motif to bind with high affinity to a totally synthetic Tat protein and to inhibit more efficiently the Tat/transactivation response element (TAR) RNA interaction as compared to the poly(I):poly(C) (pI:pC) motif. Furthermore, the pA:pU motif was tenfold more effective in inhibiting Tat-driven transactivation than the pI:pC motif. Following intranasal immunization of mice, both dsRNA motifs enhanced the antibody (serum and mucosal) and cellular responses to Tat. However, only the serum samples of mice immunized with Tat + pI:pC inhibited Tat-driven transactivation. The profile of serum antibody subclasses together with the secreted cytokines by Tat-stimulated splenocyte cultures indicated that both dsRNA motifs favored the induction of a balanced Th1 and Th2 immune response. The demonstration in this study that two dsRNA motifs had a marked effect on Tat/TAR RNA interaction and on the neutralizing capacity of anti-Tat specific antibody responses highlights their potential for biological applications and the importance of selecting the appropriate motif as an adjuvant for vaccine design. PMID- 15789359 TI - HSV induces an early primary Th1 CD4 T cell response in neonatal mice, but reduced CTL activity at the time of the peak adult response. AB - Neonates are highly susceptible to HSV. In this study, we analyzed the primary neonatal cell-mediated response to HSV at the site of T cell activation, the draining lymph nodes (LN), and examined the effects of dose and the ability of HSV to replicate on the strength and character of this response. Neonatal mice mounted a predominantly Th1 cytokine (IFN-gamma) response at all doses of a replication-competent thymidine kinase-negative HSV-2 strain (186DeltaKpn) and at high doses of a replication-defective HSV-2 virus (dl5-29, UL5(-)/UL29(-)). Both neonates and adults showed increased production of Th2 cytokines (IL-4 and/or IL 5) at high doses of the replication-defective or inactivated HSV strains. An age dependent difference in the strength of the Th1 response was noted, with neonates mounting adult-like responses at low but not high doses of HSV. Neonatal mice also showed impaired CD8(+) T cell activation and reduced HSV-specific CTL effector function at the time of the peak adult response. These studies are the first to highlight the impaired primary neonatal T cell response to HSV in the LN. PMID- 15789360 TI - Fostering coping and nurturing hope when discussing the future with terminally ill cancer patients and their caregivers. AB - BACKGROUND: It is challenging for clinicians to foster coping and allow hope when discussing prognosis and end-of-life (EOL) issues with terminally ill cancer patients and their caregivers. To the authors' knowledge, little research evidence is currently available to guide clinical practice. METHODS: The authors conducted focus groups and individual interviews with 19 patients with far advanced cancer and 24 caregivers from 3 palliative care (PC) services in Sydney, and 22 PC health professionals (HPs) from around Australia. The focus groups and individual interviews were audiotaped and fully transcribed. Additional focus groups or individual interviews were conducted until no additional topics were raised. The participants' narratives were analyzed using qualitative methodology. RESULTS: All participant groups believed there were ways of fostering coping and nurturing hope when discussing prognosis and EOL issues. Themes identified from the transcripts regarding ways of helping patients with a limited life expectancy to cope were: 1) emphasize what can be done (namely: a) control of physical symptoms; b) emotional support, care, and dignity; and c) practical support), 2) explore realistic goals, and 3) discuss day-to-day living. Two additional themes were identified regarding ways of fostering hope: the balance between truth telling and nurturing hope, and the spectrum of hope. All these themes were raised by the patients, caregivers, and HPs. However, there were some differences of opinion within the three participant groups. CONCLUSIONS: The results of the current study demonstrated that most participants believed there were ways of fostering coping and maintaining hope and that HPs have a role in facilitating this with terminally ill cancer patients and their caregivers. PMID- 15789361 TI - Mice with chimeric human livers: who says supermodels have to be tall? PMID- 15789362 TI - Overexpressed in anaplastic thyroid carcinoma-1 (OEATC-1) as a novel gene responsible for anaplastic thyroid carcinoma. AB - BACKGROUND: Anaplastic thyroid carcinoma (ATC) is one of the most fulminant human malignancies. However, the molecular carcinogenic mechanisms of ATC are understood poorly. Recently, the authors performed a cyclic DNA (cDNA) microarray analysis with 11 anaplastic thyroid carcinoma cell lines (ACLs) and discovered several novel responsible genes for ACLs and ATC. From the extended list, they focused on hypothetical and anonymous genes and investigated a novel gene, named the overexpressed in anaplastic thyroid carcinoma-1 (OEATC-1) gene. METHODS: To investigate the role of the OEATC-1 gene in ATC carcinogenesis, first, the expression levels of OEATC-1 in ACLs, in various types of carcinoma cell lines, and in normal human tissues were examined with reverse transcriptase-polymerase chain reaction analysis. To explore the effect of OEATC-1 in ATC development, a cell-growth assay was performed with KTA2 cells under OEATC-1 gene silencing using small-interfering RNA (siRNA). RESULTS: OEATC-1 was overexpressed significantly in ACLs and in other types of carcinoma cell lines with various expression levels. Conversely, in normal human tissues, OEATC-1 was expressed weakly in placenta, kidney, spleen, thymus, small intestine, and thyroid gland. To evaluate the effects of OEATC-1 on tumor cell growth, gene silencing was caused by transfecting the plasmid-generating siRNA effect to KTA2 cells. Consequently, the silencing of OEATC-1 significantly suppressed the cell growth compared with controls. CONCLUSIONS: The current results indicated that OEATC-1 may have some oncogenic or cell growth-promoting function in ACL. OEATC-1 is considered a novel responsible gene in ATC. PMID- 15789363 TI - The needs of terminally ill cancer patients versus those of caregivers for information regarding prognosis and end-of-life issues. AB - BACKGROUND: The difficulty of negotiating the concerns of family members while also respecting the needs of the patient adds complexity to the task of discussing prognosis and end-of-life (EOL) issues with terminally ill cancer patients. The informational needs of caregivers may be different from those of the patients themselves with regard to these topics. However, to the authors' knowledge, this issue has received relatively little research attention. METHODS: The authors conducted focus groups and individual interviews with 19 patients with far advanced cancer and 24 caregivers from 3 palliative care (PC) services in Sydney and 22 PC health professionals (HPs) from around Australia. The focus groups and individual interviews were audiotaped and fully transcribed. Additional focus groups or individual interviews were conducted until no additional topics were raised. The participants' narratives were analyzed using qualitative methodology. RESULTS: The participants had varying views regarding whether patients and caregivers should be told different information concerning prognosis and EOL issues. Three themes were identified from the transcripts regarding meeting the informational needs of both the patients and caregivers: 1) the importance of consistency and openness, 2) the specific information needed to care for the patient, and 3) the value of having separate discussions with the patient and caregiver. A desire to restrict the patient's access to information by the caregiver or vice versa was reported by the HPs to be one of the most challenging issues when discussing prognosis and EOL issues. Three themes were identified with regard to this issue: 1) autonomy versus protection, 2) negotiating family dynamics, and 3) difficulty using interpreters. CONCLUSIONS: The results of the current study emphasized the importance of considering the distinct informational needs of caregivers, as well as those of the patient, when discussing prognosis and EOL issues. PMID- 15789365 TI - Regulatory T cells and the liver: a new piece of the puzzle. PMID- 15789364 TI - Oral treatment of the TRAMP mice with doxazosin suppresses prostate tumor growth and metastasis. AB - BACKGROUND: We used the TRAMP mouse model for testing the effect of oral doxazosin treatment on the in vivo prostate tumor growth and metastasis. METHODS: Five groups of TRAMP mice at different ages were orally fed with 1 mg/kg of doxazosin or DMSO for 45-196 days. At the end of oral treatment, tumor weight was determined, and metastasis to multiple organs examined. The levels of MUC18, Bcl 2, Bax, caspase-3, poly (ADP-ribose) polymerase (PARP), phospho (Ser473)-AKT, and Ki-67 in the mouse prostate tumors were determined. RESULTS: Oral treatment of the TRAMP mice with doxazosin for 45-81 days did not decrease the size of preexisting prostate tumors, but it limited the metastasis to peri-aortic lymph nodes. A prolonged treatment of TRAMP mice with doxazosin (156-196 days), if administered early, decreased the prostate tumor weight and completely suppressed metastasis. The doxazosin treatment did not further decrease the expression of an already low level of Bcl-2 in all prostate tumors, but it increased the expression of Bax, and the activation of caspase-3, and the cleavage of a downstream substrate, PARP. The treatment reduced the expression of MUC18, phospho (Ser473)-AKT, and Ki-67. The treatment in the early phase appeared to promote prostate tumor growth and increased the expression of a proliferative index, Ki-67. CONCLUSIONS: Doxazosin, if administered early, may be useful for preventing the prostate tumor formation, and also for limiting or completely suppressing the metastasis of prostate cancer in the TRAMP model. The mechanism of doxazosin is consistent with the established hypothesis. PMID- 15789366 TI - Temporal and spatial distribution of growth-associated molecules and astroglial cells in the rat corticospinal tract during development. AB - To understand better the role of growth-promoting and -inhibiting molecules in the development of the corticospinal tract (CST), temporospatial expression of embryonic neural cell adhesion molecule (E-NCAM), growth-associated protein-43 (GAP-43), and chondroitin sulfate proteoglycan (CSPG) was studied in developing rats. Transverse sections of the seventh cervical (C7), seventh thoracic (T7), and fourth lumbar (L4) segments were examined at postnatal days (P) 2, 6, 10, 14, and 28. The highest E-NCAM immunoreactivity appeared at the C7 level on P2 and shifted caudally to the T7 on P6 and L4 on P10, which correlated closely with the time course of CST development. The peak expression of GAP-43 emerged at C7 on P2 and shifted to the T7 and L4 levels at a relatively lagging pace compared with that of E-NCAM. Conversely, a transient reduction in CSPG immunoreactivity was found within the CST at the C7 level on P2, T7 level on P6, and L4 level on P10, corresponding well with the arrival of CST-leading axons at these levels. Interestingly, higher levels of CSPG were found to surround the growing CST, suggesting a repulsive environment that channels the growth of CST. Moreover, a transition from immature to mature astrocytes in a rostrocaudal direction during CST development was evidenced by anti-vimentin and anti-glial fibrillary acidic protein (GFAP) immunostaining, suggesting a guidance role of immature astroglia in axonal outgrowth. Our study thus demonstrated dynamic changes of multiple growth-related molecules and astroglial environment that contribute to postnatal development of the CST. PMID- 15789367 TI - Statins and hepatotoxicity: focus on patients with fatty liver. PMID- 15789368 TI - Acute liver failure in children: the role of defects in fatty acid oxidation. PMID- 15789369 TI - Activation of extracellular signal-regulated kinase 5 may play a neuroprotective role in hippocampal CA3/DG region after cerebral ischemia. AB - Extracellular signal-regulated kinase 5 (ERK5), the newest member of the mitogen activated protein (MAP) kinase family of proteins, is widely expressed in many tissues, including the brain. Here we investigated the activation and subcellular localization of ERK5 by immunoblotting and immunohistochemistry as well as its potential role following cerebral ischemia in rat hippocampus. Transient cerebral ischemia was induced by the four-vessel occlusion method in Sprague-Dawley rats. Our results first indicated that the strongly activated ERK5 immunoreactivity was seen in the CA3/DG region but not in the CA1 pyramidal cell of rat hippocampus following reperfusion. In cytosol extracts, ERK5 activation was rapidly increased, with a peak at 30 min, and then gradually decreased to basal level at 3 days of reperfusion. In nucleus extracts, both phospho-ERK5 and its protein expression were persistently enhanced during the later reperfusion period (from 6 hr to 3 days). To elucidate further the possible role of ERK5 activation and subcellular localization in ischemic insult, rats were intraperitoneally administrated with nifedipine (ND) and dextromethorphan (DM), inhibitors of two types of calcium channels, 20 min prior to ischemia. Our findings showed that ND or DM significantly reduced activated ERK5 immunoreactivity in the nucleus and that most of the CA3/DG neurons were lost 3 days later. Most importantly, intracerebroventricular infusion of ERK5 antisense oligonucleotides (AS; every 24 hr for 3 days before ischemia), but not sense oligonucleotides or vehicle, not only markedly decreased the level of ERK5 and p-ERK5 but also largely caused neuronal loss in the CA3/DG region at 3 days of reperfusion. Taken together, the results strongly suggest that ERK5 was selectively activated in the hippocampal CA3/DG region and subsequently translocated from the cytosol to the nucleus through activation of N-methyl-D-aspartate receptor and L-type voltage-gated calcium channel, which might act as an important survival signal in ischemia induced neuronal cell damage of the CA3/DG region. PMID- 15789372 TI - Effect of nisin on the growth of Staphylococcus aureus determined by a microcalorimetric method. AB - A novel microcalorimetric technique based on the bacterial heat output was applied to evaluate the biological effect of nisin on the growth of Staphylococcus aureus. The thermogenic curves of S. aureus in the presence of nisin were studied by an LKB-2277 Thermal Activity Monitor. The thermokinetic parameters, such as the growth rate constant (k), the generation times (G), the inhibitory ratio (I), and the half inhibitory concentration (IC50), for the growth of S. aureus at different nisin concentrations were determined. The relationship between the growth rate constant (k) and the concentration of nisin (c) is nearly linear, which can be modeled by the formula k = 0.03794 - 4.005 x 10(-4) x c, with a correlation coefficient of -0.9971. Based on this model, we obtained the critical inhibitory concentration of nisin on the growth of S. aureus at 94.73 IU/mL. We proposed that this microcalorimetric method could be a useful tool in monitoring the biological effect of nisin on microorganisms, and providing valuable information on the study of microorganism metabolisms. PMID- 15789371 TI - Elimination of Escherichia coli O 157 : H7 and Listeria monocytogenes from raw beef sausage by gamma-irradiation. AB - The effectiveness of low gamma-irradiation doses in the destruction of Escherichia coli O 157 : H7 and Listeria monocytogenes in raw beef sausages was investigated. Raw samples of fresh manufactured beef sausage were subjected to gamma-irradiation at doses of 0, 1, 2, and 3 kGy. Then samples were cold-stored (4 +/- 1 degrees C) for 12 days and the effects of irradiation and storage on the counts of these harmful bacteria were studied. Moreover, the effects of irradiation and storage on the percentages of free fatty acids (FFAs) in lipids, on the p-anisidine values of lipids, solubility of sarcoplasmic and myofibrilar proteins, and water-holding capacity (WHC) were also determined. The results showed that gamma-irradiation at 1 and 2 kGy significantly reduced the counts of E. coli O 157 : H7 and L. monocytogenes, while 3 kGy dose effectively eliminated these bacteria by more than 4 log and 3 log units, respectively, and could keep their counts below the detection level during storage. Gamma-irradiation had no significant effects on the percentages of FFAs in lipids, solubility of sarcoplasmic and myofibrilar proteins, and WHC of samples, while it significantly increased the p-anisidine value of lipids. During storage, significant increases in the percentages of FFAs and p-anisidine values were observed for lipids of irradiated and nonirradiated sausages, while the solubility of sarcoplasmic and myofibrilar proteins showed no significant changes. Moreover, samples of irradiated and nonirradiated sausages showed significant decreases in their WHC during the first 6 days of storage at 4 +/- 1 degrees C, then showed no significant changes. Finally, gamma-irradiation at a dose of 3 kGy appeared to be sufficient to improve the microbiological safety of raw beef sausages without adverse effects on their chemical properties. PMID- 15789375 TI - The prevalence and clinical significance of amniotic fluid 'sludge' in patients with preterm labor and intact membranes. AB - OBJECTIVE: To determine the prevalence and clinical significance of amniotic fluid (AF) 'sludge' observed during transvaginal ultrasound examination of the cervix in patients with preterm labor and intact membranes, and in those with uncomplicated pregnancies. METHODS: This retrospective study included patients with preterm labor and intact membranes (n = 84) and those with uncomplicated term pregnancies (n = 298). The outcome variables included the occurrence of documented microbial invasion of the amniotic cavity (MIAC), histological chorioamnionitis, examination-to-delivery interval, admission to the neonatal intensive care unit (NICU), a composite neonatal morbidity, perinatal death, and delivery within 48 h, 7 days, and < 35 weeks and < 32 weeks. Statistical analysis included Chi-square test, stepwise logistic regression analysis and survival analysis. RESULTS: The prevalence of AF 'sludge' was 1% (3/298) in patients with uncomplicated term pregnancies and 22.6% (19/84) in those with preterm labor and intact membranes. Among patients with preterm labor and intact membranes: (1) cervical length < or = 15 mm was present in 58.3% (49/84) of the patients; (2) the prevalence of MIAC and histological chorioamnionitis was 12.1% (7/58) and 32.9% (25/76), respectively; (3) the rate of spontaneous preterm delivery within 48 h, 7 days, and < 32 weeks and < 35 weeks of gestation was 13.6% (8/59), 28.8% (17/59), 39.5% (17/43) and 50.8% (30/59), respectively; (4) patients with AF 'sludge' had a higher frequency of positive AF cultures [33.3% (6/18) vs. 2.5% (1/40), P = 0.003] and histological chorioamnionitis [77.8% (14/18) vs. 19% (11/58), P < 0.001] than those without AF 'sludge'; (5) a higher proportion of neonates born to patients with AF 'sludge' was admitted to the NICU [64.3% (9/14) vs. 12.9% (8/62), P < 0.01], had a composite neonatal morbidity [36.8% (7/19) vs. 13.8% (9/65), P = 0.04] and died in the perinatal period [36.8% (7/19) vs. 4.6% (3/65), P = 0.001] than those born to women without 'sludge'; (6) a higher proportion of patients with AF 'sludge' had spontaneous delivery within 48 h [42.9% (6/14) vs. 4.4% (2/45), P = 0.001], within 7 days [71.4% (10/14) vs. 15.6% (7/45), P < 0.001], < 32 weeks [75% (9/12) vs. 25.8% (8/31), P = 0.005] and < 35 weeks [92.9% (13/14) vs. 37.8% (17/45), P < 0.001] than those without AF 'sludge'; and (7) patients with AF 'sludge' had a shorter examination-to-delivery interval than those without AF 'sludge' [AF 'sludge' median, 1 (IQR, 1-5) days vs. no AF 'sludge' median, 33 (IQR, 18-58) days; P < 0.001]. CONCLUSION: The presence of AF 'sludge' in patients with preterm labor and intact membranes is a risk factor for MIAC, histological chorioamnionitis and impending preterm delivery. PMID- 15789376 TI - End binding protein-1 (EB1) complements microtubule-associated protein-1B during axonogenesis. AB - The different strains of microtubule-associated protein (MAP)-1B-deficient mice that have been generated appear to express different phenotypes. This variability could be the consequence of the distinct genetic backgrounds of the animals used to generate these lines. Certain proteins might be able to complement the deficit of MAP1B function in these mice. Therefore, we examined whether the concentrations of potential compensatory proteins varied among these mutant strains. In this way, we identified significant differences in the amounts of the microtubule-associated EB1 protein between two of these strains. Furthermore, in transfection studies, we demonstrated that the overexpression of end binding protein-1 (EB1) could facilitate axonogenesis in MAP1B-/- cells in which EB1 is normally weakly expressed. Thus, we suggest that EB1 could complement MAP1B function during neural development. PMID- 15789378 TI - Pericardiocentesis ending in the superior vena cava. AB - Pericardiocentesis is associated with a wide range of complications. We describe a pericardiocentesis attempt ending in the superior vena cava through the route of liver parenchyma, hepatic vein, inferior vena cava, and right atrium. PMID- 15789377 TI - Activation of gamma-aminobutyric acid GAT-1 transporters on glutamatergic terminals of mouse spinal cord mediates glutamate release through anion channels and by transporter reversal. AB - The effects of gamma-aminobutyric acid (GABA) on the release of glutamate from mouse spinal cord nerve endings have been studied using superfused synaptosomes. GABA elicited a concentration-dependent release of [3H]D-aspartate ([3H]D-ASP; EC50= 3.76 microM). Neither muscimol nor (-)baclofen mimicked GABA, excluding receptor involvement. The GABA-evoked release was strictly Na+ dependent and was prevented by the GABA transporter inhibitor SKF89976A, suggesting involvement of GAT-1 transporters located on glutamatergic nerve terminals. GABA also potentiated the spontaneous release of endogenous glutamate; an effect sensitive to SKF89976A and low-Na+-containing medium. Confocal microscopy shows that the GABA transporter GAT-1 is coexpressed with the vesicular glutamate transporter vGLUT-1 and with the plasma membrane glutamate transporter EAAT2 in a substantial portion of synaptosomal particles. The GABA effect was external Ca2+ independent and was not decreased when cytosolic Ca2+ ions were chelated by BAPTA. The glutamate transporter blocker DL-TBOA or dihydrokainate inhibited in part (approximately 35%) the GABA (10 microM)-evoked [3H]D-ASP release; this release was strongly reduced by the anion channel blockers niflumic acid and NPPB. GABA, up to 30 microM, was unable to augment significantly the basal release of [3H]glycine from spinal cord synaptosomes, indicating selectivity for glutamatergic transmission. It is concluded that GABA GAT-1 transporters and glutamate transporters coexist on the same spinal cord glutamatergic terminals. Activation of these GABA transporters elicits release of glutamate partially by reversal of glutamate transporters present on glutamatergic terminals and largely through anion channels. PMID- 15789379 TI - Stenting vs. balloon angioplasty for discrete unoperated coarctation of the aorta in adolescents and adults. AB - More information is needed to clarify whether stenting is superior to balloon angioplasty (BA) for unoperated coarctation of the aorta (CoA). From September 1997, 21 consecutive adolescents and adults (24 +/- 11 years) with discrete CoA underwent stenting (G1). The results were compared to those achieved by BA performed in historical group of 15 patients (18 +/- 10 years; P = 0.103; G2). After the procedure, systolic gradient reduction was higher (99% +/- 2% vs. 87% +/- 17%; P = 0.015), residual gradients lower (0.4 +/- 1.4 vs. 5.9 +/- 7.9 mm Hg; P = 0.019), gain at the CoA site higher (333% +/- 172% vs. 190% +/- 104%; P = 0.007), and CoA diameter larger (16.9 +/- 2.9 vs. 12.9 +/- 3.2 mm; P < 0.001) in G1. Aortic wall abnormalities were found in eight patients in G2 (53%) and in one in G1 (7%; P < 0.001). There was no major complication. Repeat catheterization (n = 33) and/or MRI (n = 2) was performed at a median follow-up of 1.0 year for G1 and 1.5 for G2 (P = 0.005). Gradient reduction persisted in both groups, although higher late gradients were seen in G2 (median of 0 mm Hg for G1 vs. 3 for G2; P = 0.014). CoA diameter showed no late loss in G1 and a late gain in G2 with a trend to being larger in G1 (16.7 +/- 2.9 vs. 14.6 +/- 3.9 mm; P = 0.075). Two patients required late stenting due to aneurysm formation or stent fracture in G1. Aortic wall abnormalities did not progress and one patient required redilation in G2. Blood pressure was similar in both groups at follow-up (126 +/- 12/81 +/- 11 for G1 vs. 120 +/- 15/80 +/- 10 mm Hg for G2; P = 0.149 and 0.975, respectively). Although satisfactory and similar clinical outcomes were observed with both techniques, stenting was a better means to relieve the stenosis and minimize the risk of developing immediate aortic wall abnormalities. PMID- 15789381 TI - Percutaneous closure of left ventricular-to-right atrial fistula after prosthetic mitral valve rereplacement using the Amplatzer duct occluder. AB - A 70-year-old female with a history of rheumatic heart disease underwent rereplacement of mitral valve mechanical prosthesis in May 2003. Seven months later, she presented with progressive exertional dyspnea, exercise intolerance, and a new holosystolic/diastolic murmur. Echocardiography confirmed a large shunt through a fistula in the inferior limbus of the atrial septum with left ventricular-to-right atrial communication. We report the novel use of the Amplatzer duct occluder for closure of the fistulous tract. PMID- 15789382 TI - Renal fractional flow reserve: a hemodynamic evaluation of moderate renal artery stenoses. AB - The objective of this study was to perform a hemodynamic evaluation of moderate (50-90%) renal artery stenosis (RAS) under conditions of rest and maximum hyperemia. Identifying patients with RAS who have hemodynamically significant stenoses and are most likely to benefit from revascularization is clinically important. Current methods used to evaluate RAS, including angiography, have limitations. Physiologic evaluation of RAS may have a role in identifying patients with hemodynamically significant stenosis. Patients with suspected renovascular hypertension due to aorto-ostial RAS were included in the study. Hyperemia was induced by administration of intrarenal papavarine. Translesional pressure gradients were measured and renal fractional flow reserve (FFR) was calculated using a 0.014'' pressure guidewire. Thirteen patients and 14 arteries with moderately severe (50-90%) RAS were studied. The mean translesional pressure gradient rose from a baseline of 6.3 +/- 3.9 to 17.5 +/- 10.8 mm Hg with maximal hyperemia. The renal FFR ranged from 0.58 to 0.95. There was a poor correlation between angiographic stenosis measurement and the renal FFR (r = -0.18; P = 0.54) and the hyperemic translesional mean pressure gradient (r = 0.22; P = 0.44). There was an excellent correlation between renal FFR and the resting mean translesional pressure gradient (r = -0.76; P = 0.0016) and the hyperemic mean translesional pressure gradient (r = -0.94; P < 0.0001). Selective renal arterial papavarine administration induces maximum hyperemia, permitting the calculation of renal FFR in renal arteries with aorto-ostial stenoses. The renal FFR correlates well with other hemodynamic parameters of lesion severity, but poorly with angiographic measures of lesion severity. PMID- 15789383 TI - Mitral stenosis in young children: treatment without surgery. PMID- 15789384 TI - Treating chronic total occlusions using subintimal tracking and reentry: the STAR technique. AB - Successful recanalization of coronary total occlusions (CTOs) remains an area where improvements are needed. We propose an approach similar to the one utilized in treating some peripheral artery occlusions and aimed to create a subintimal dissection with distal reentry. A 0.014' hydrophilic wire with a J-configuration is utilized for this purpose. We applied this technique to CTO of native coronaries in 31 patients where previous attempts failed in 21 of them (67%). The right coronary artery (RCA) was the index vessel in 87% of patients. Recanalization of the vessel and of most of distal branches was achieved in 21 patients; patency of at least one major distal branch was achieved in 9 patients. Drug-eluting stents (DESs) were implanted in 53% of patients. Three patients had intraprocedural vessel perforation without consequences. Five patients (16%) had in-hospital non-Q-wave myocardial infarction. No other adverse events occurred at a mean follow-up of 5.1 +/- 3.7 months except for one noncardiac death. Angiographic follow-up was performed in 21 (67%) patients and 53% of them developed restenosis. Reintervention on the target vessel was performed in 11 patients (52%). Complete success with the procedure was originally obtained in 8 of the 10 patients who did not develop restenosis and in 8 of them DESs were originally implanted. This technique appears a promising approach to recanalize difficult total occlusions, particularly the ones localize on the RCA, which has the most important side branches located distally. PMID- 15789386 TI - Use of cutting balloon for palliative treatment in tetralogy of Fallot. AB - In tetralogy of Fallot, severe cyanosis due to insufficient pulmonary blood flow necessitates early intervention. The Cutting Balloon, developed for percutaneous coronary angioplasty, was used successfully for transcatheter incision and dilation of infundibular and valvar pulmonary stenosis in four patients with tetralogy of Fallot. PMID- 15789385 TI - Efficacy and safety of oral sirolimus to inhibit in-stent intimal hyperplasia. AB - Sirolimus systemic administration has shown marked inhibition of neointimal hyperplasia (NIH) after balloon angioplasty in porcine models. In this pilot study, we tested the hypothesis that oral sirolimus is safe and effective to inhibit in-stent NIH and therefore to prevent and treat in-stent restenosis (ISR). Twelve patients (18 lesions) with high risk for ISR, including 8 ISR lesions, were admitted. One day before the procedure, patients were given a 15 mg loading dose of oral sirolimus, followed by 5 mg daily for 28 days, with weekly whole blood level measurements. The daily dose was adjusted to keep the concentration at 10-15 ng/ml. Sirolimus was well tolerated by all patients but one, who died at the end of the third week of treatment. The 4- and 8-month follow-up revealed an angiographic late loss of 0.40 +/- 0.24 and 0.67 +/- 0.45 mm (P < 0.01), respectively. At the same time points, the intravascular ultrasound in in-stent relative volumetric obstruction was 14.4% +/- 9.1% and 23.2% +/- 10.1% (P < 0.01), respectively. At 24-month clinical follow-up, adverse events were one (8.3%) death, two (11.1%) target lesion, and four (22.2%) target vessel revascularizations. In conclusion, in this small group of high-risk ISR patients, oral sirolimus inhibited NIH and therefore may be an effective strategy for the prevention and treatment of ISR. PMID- 15789387 TI - Closure of a moderately large atrial septal defect with a self-fabricated fenestrated Amplatzer septal occluder in an 85-year-old patient with reduced diastolic elasticity of the left ventricle. AB - Percutaneous closure of an atrial septal defect (ASD) in the elderly with reduced diastolic elasticity of the left ventricle poses a significant management challenge. We report on the case of an 85-year-old patient who was admitted for percutaneous device closure of a moderately large secundum atrial septal defect. Hemodynamic evaluation documented an increase in left atrial pressure from a mean of 12 mm Hg to a mean of 32 mm Hg after balloon test occlusion of the ASD. Two months later, after adequate pretreatment with diuretics and afterload-reducing substances, he underwent successful closure of the ASD using a self-fabricated fenestrated Amplatzer septal occluder, which resulted in a postimplantation left atrial pressure of a mean of 18 mm Hg. Recovery was unremarkable and the fenestration has remained patent for 3 months since implantation of the device. This unique case highlights the feasibility of using a self-fabricated fenestrated Amplatzer septal occluder to close interatrial communications in elderly patients with diastolic dysfunction of the left ventricle. PMID- 15789388 TI - N-acetylcysteine prophylaxis significantly reduces the risk of radiocontrast induced nephropathy: comprehensive meta-analysis. AB - The objectives of this study was to assess the overall effect of N-acetylcysteine (NAC) in preventing radiocontrast-induced nephropathy (RCIN) using all available data in the literature. RCIN is associated with increased morbidity and mortality. Existing randomized trials of NAC are small and show inconsistent results. Prior meta-analyses do not include data from the most current studies. We used standard search protocols to identify all published articles and abstracts of prospective trials using NAC with fluid hydration compared to hydration alone in patients with chronic renal insufficiency undergoing contrast procedures. A rise in serum creatinine by 0.5 mg/dl or 25% above baseline at 48 72 hr after contrast exposure was used as the primary outcome. We identified 14 trials of NAC with 1,584 patients published as full-text articles. Using a random effects model, the use of oral NAC resulted in a significant reduction in the risk for developing RCIN (RR = 0.57; 95% CI = 0.37-0.84; P = 0.01). This finding did not significantly change in a fixed-effect model (RR = 0.55; 95% CI = 0.42 0.73) or when the data were reanalyzed using only randomized trials in all forms (i.e., articles and abstracts; RR = 0.67; 95% CI = 0.47-0.95). We identified only one important difference between the positive and the negative studies: the cumulative exposure to contrast media (174 vs. 152 ml). Metaregression did not show a significant relationship between contrast volume and the RR of developing RCIN (P > 0.10). In the trials showing benefit for NAC, the treated patients' postprocedure creatinine unexpectedly decreased by 0.21 mg/dl (95% CI = 0.33 0.08). Prophylaxis with NAC significantly reduces the risk for RCIN. The reasons for improvement in serum creatinine in patients treated with NAC are unclear, but may include improved renal blood flow due to NAC and/or vigorous hydration. PMID- 15789389 TI - Intermediate-term results of percutaneous transvenous mitral commissurotomy in children less than 12 years of age. AB - The objectives of this study were to review the efficacy of percutaneous transvenous mitral commissurotomy (PTMC) in young children less than 12 years of age and to provide intermediate-term follow-up data. There is a paucity of information regarding the long-term efficacy of PTMC done in children less than 12 years of age. The data of 100 consecutive children less than 12 years of age (mean, 11.1 +/- 1.2 years) who underwent PTMC using Inoue balloon were analyzed retrospectively. Serial clinical and echocardiographic follow-up information of more than 6 months was available in 94 patients. The procedure was successful in 94 patients. The mean calculated mitral valve area (MVA) increased from 0.72 0.14 to 1.7 0.35 cm(2) (P 0.0001). Echocardiographic restenosis (MVA < or = 1 cm(2) or > 50% gain loss) occurred in 14 of 94 patients (16%) over a mean follow-up of 34.4 25.9 (range, 2-115) months. The improvement in New York Heart Association (NYHA) functional class was maintained in most patients (from a mean of 2.87 0.5 pre-PTMC to 1.42 0.6 at follow-up). Seven out of 14 patients with restenosis underwent a re-PTMC. The actuarial rate of good functional status (survival, no repeat interventions, and NYHA class 1 or 2) at 100 months was 75.4% 8.7%. PTMC provides excellent intermediate-term palliation even in young children with rheumatic mitral stenosis. PMID- 15789391 TI - Impact of frame selection on quantitative coronary angiographic analysis after coronary stenting. AB - This study examined the impact of frame selection on the results of quantitative coronary angiographic analysis (QCA). Thirty-nine patients who had stent implantation and a follow-up angiogram 5-7 months later were studied using the CMS Medis QCA system. The acute and follow-up films (n = 39) were read in three different ways to assess the impact of frame selection: frame chosen making the stent appear least narrowed (best); frame chosen making the stent appear most narrowed (worst); and measurement from the mean value from three consecutive end diastolic frames (core). We measured the mean percent diameter stenosis immediately postintervention and at follow-up, the binary restenosis rate, and the late lumen loss (mm). There was a statistically significant difference in all of these variables when comparing the three methods of frame selection (best vs. worst, P values < 0.001; best or worst vs. core, P values < 0.01). This study demonstrates a marked variability in the results obtained using QCA to measure the acute and late coronary stent outcomes when operators have the ability to select which frame to analyze (frame bias). PMID- 15789392 TI - Another attempt to deal with restenosis. PMID- 15789393 TI - Outpatient coronary angioplasty: feasible and safe. AB - This study tested the safety and feasibility of coronary angioplasty on an outpatient basis. The purpose of this approach includes cost-effectiveness and patient comfort. Six hundred forty-four patients were randomized to either transradial or transfemoral PTCA using 6 Fr equipment. Patients were triaged to outpatient management based on a predefined set of predictors of an adverse outcome in the first 24 hr after initially successful coronary angioplasty. Three hundred seventy-five patients (58%) were discharged 4-6 hr after PTCA; 42% stayed in hospital overnight. In the outpatient group, one adverse event occurred (subacute stent thrombosis 7 hr postdischarge, nonfatal myocardial infarction). There were no major vascular complications. In the hospital group, 19 patients (7%) sustained an adverse cardiac even in the first 24 hr; 1 patient died. Patients treated via the femoral route had more (minor) bleeding complications (19 patients; 6%); in 17 of these, this was the sole reason that discharge was delayed. PTCA on an outpatient basis, performed via the radial or the femoral artery with low-profile equipment, is safe and feasible in a considerable part of a routine PTCA population. A larger proportion of transradial patients can be discharged due to a reduction in (minor) bleeding complications. PMID- 15789395 TI - Effect of continuous quality improvement analysis on the delivery of primary percutaneous revascularization for acute myocardial infarction: a community hospital experience. AB - As time to reperfusion correlates with outcomes, a door-to-balloon time of 90 +/- 30 min for primary percutaneous coronary revascularization (PCI) for the treatment of acute myocardial infarction has been recently established as a guideline by the ACC/AHA. The purpose of this study is to assess the effects of a continuous quality assurance program designed to expedite primary angioplasty at a community hospital. A database of all primary PCI procedures was created in 1998. Two groups of consecutive patients treated with primary PCI were studied. Group 1 represented patients in the time period between 1 June 1998 to 1 November 1998 and group 2 represented patients in the period between 1 January 2000 and 16 June 2000. Continuous quality assurance analysis was performed. Modifications to the primary angioplasty program were initiated in the latter group. Time intervals to certain treatment landmarks were compared between the groups. Significant decreases in the time intervals from emergency room registration to initial electrocardiogram (8.4 +/- 8.2 vs. 3.7 +/- 19.5 min; P < 0.001), presentation to the catheterization laboratory to arterial access (13.5 +/- 12.9 vs. 11.6 +/- 5.8 min; P < 0.001), and emergency room registration to initial angioplasty balloon inflation (132.0 +/- 69.2 vs. 112 +/- 72.0 min; P < 0.001) were achieved. For the subgroup of patients presenting with diagnostic ST elevation myocardial infarction, a large decrease in the door-to-balloon time interval between group 1 and group 2 was demonstrated (114.15 +/- 9.67 vs. 87.92 +/- 10.93 min; P = NS), resulting in compliance with ACC/AHA guidelines. Continuous quality improvement analysis can expedite care for patients treated by primary PCI in the community hospital setting. PMID- 15789396 TI - Changing paradigm for neointimal cell origin: is restenosis a blood-borne disease? PMID- 15789397 TI - Drug eluting stents: case study in the acceptance of a new medical technology. PMID- 15789399 TI - Registrying knowledge: it isn't always enough. PMID- 15789398 TI - Renal failure after percutaneous coronary intervention is associated with high mortality. AB - Renal failure is a marker of poor outcome in the general population. Renal failure after percutaneous coronary artery intervention (PCI) is associated with an increased hazard of in-hospital mortality. We hypothesized that post-PCI renal insufficiency would be a predictor of long-term mortality in patients undergoing PCI who survive for over 30 days after the procedure. A retrospective analysis was conducted from a registry of 9,067 patients undergoing PCI at our center from 1997 to 2001. A rise in creatinine by 1 mg/dl from baseline was defined as post PCI renal insufficiency. Vital status was assessed using Social Security Death Index. There were a total of 996 deaths over a mean follow-up period of 3.2 years. In a multivariate analysis, history of recent acute myocardial infarction, older age, insulin-dependent diabetes, baseline creatinine greater than 1.5 mg/dl, and presence of mitral regurgitation were associated with post-PCI renal insufficiency. Developing post-PCI renal insufficiency was associated with a 4.31 fold hazard of mortality in univariate analysis and a 1.77-fold hazard after adjustment for known predictors of mortality after PCI. The 1-year survival in patients with renal failure was 70.3% +/- 3.91%, compared to a survival of 93.6% +/- 0.27% in those without any post-PCI renal insufficiency (P < 0.0001). Acute renal insufficiency after PCI is a strong and independent predictor of long-term mortality in patients who survived for 30 days after the procedure. PMID- 15789401 TI - Results and follow-up after implantation of four or more sirolimus-eluting stents in the same patient. AB - The aim of this study was to assess the safety and effectiveness of > or =4 sirolimus-eluting stent (SES; Cypher, Cordis, Johnson and Johnson) implantation. The safety of implantation of > or =4 SESs in the same patient and setting has not been established. Furthermore, it has been hypothesized that sirolimus administration with the use of multiple stents may diminish the platelet inhibitory effects of clopidogrel and may trigger drug-drug interactions. We identified 96 consecutive patients (96 procedures) who underwent implantation of > or =4 SESs in 365 lesions (438 stents) during the same procedure. All patients received aspirin indefinitely and clopidogrel or ticlopidine for at least 1 year postprocedure; 57% and 47% of the patients were on calcium channel blocker and statin therapy, respectively. All stents were successfully deployed and glycoprotein IIb/IIIa inhibitors were used in 50% of the procedures. There were no in-hospital deaths, Q-wave myocardial infarction (MI), urgent bypass surgery, or repeat percutaneous coronary intervention; 18 patients (19%) suffered non-Q wave MI (defined as CK-MB elevation >3 times the upper limit of normal). At 30 day follow-up, there was one (1%) subacute thrombosis resulting in target lesion revascularization. At mean follow-up time of 15.4 +/- 6.2 months, the frequency of target lesion revascularization, target vessel revascularization, and major adverse cardiac event rates were 12%, 16%, and 18%, respectively. No other notable clinical events that might have been attributed to the possible drug-drug interactions or discontinuation of concomitant antithrombotic, statin, or calcium channel blocker therapy were reported. Multiple (> or =4) SES implantation appears safe with no increase in major adverse cardiac events. PMID- 15789402 TI - Assembly factors of F1FO-ATP synthase across genomes. AB - Work with respiration-deficient strains of Saccharomyces cerevisiae has provided evidence that assembly of the mitochondrial ATP synthase is dependent on proteins that serve substrate-specific, chaperone-type functions: Atp10p, Atp11p, Atp12p, Atp22p, and Fmc1p. Atp11p and Atp12p mediate the formation of the F1 moiety via interaction with subunits F1-beta and F1-alpha, respectively. The role of Fmc1p is less clear. Atp10p and Atp22p are essential for the formation of the F(O) part, during which Atp10p assists in the incorporation of the F(O)-a subunit. Here we present a comprehensive analysis of ATP synthase assembly factors from all available genomes. The mechanism of the F1 assembly is preserved in all eukaryotic lineages that are capable of ATP synthesis via oxidative phosphorylation and requires Atp11p and Atp12p. Conversely, composition of the F(O) part as well as its assembly is more versatile. We found two distinct subtypes of the F(O)-a subunit, one of which seems to be dependent on the action of Atp10p while the other does not. Restricted occurrence of Fmc1p and Atp22p suggests the existence of lineage-specific assembly factors. Our phylogenetic data served as a source for comparative sequence analysis, which identified evolutionarily conserved residues, putative functional domains and their basic structural features for Atp10p, Atp11p, and Atp12p orthologs. These results provide the basis for detailed molecular analysis of the ATP synthase-specific chaperones. PMID- 15789403 TI - S-phase entry of oligodendrocyte lineage cells is associated with increased levels of p21Cip1. AB - The mechanisms regulating the number of myelinating cells in the central nervous system are crucial for both normal development and repair in pathological conditions. Among relevant growth factors involved in this process, fibroblast growth factor-2 (FGF2) induces oligodendrocyte progenitors (OLPs) to proliferate and stimulates mature oligodendrocytes (OLs) to reenter the S-phase of the cell cycle. S-phase entry is modulated by the formation of complexes between cyclins and cyclin-dependent kinases (CDKs), on one hand, and by their interactions with cell cycle inhibitors (e.g., p18INK, p27Kip1, p21Cip1), on the other. Although the roles of cyclin E/CDK2 complexes and the inhibitor p27Kip1 have been extensively investigated relative to proliferation and differentiation in the OL lineage, less is known about the regulation of the formation of cyclin D1/CDK4 complexes and the role of p21Cip1 in these events. In this study, we show that the FGF2-mediated increase in bromodeoxyuridine (BrdU) incorporation into OL progenitors and mature OLs occurs concomitantly with increase in the levels of p21Cip1 and the formation of p21Cip1/cyclin D1/CDK4 ternary complexes. These complexes are functionally active is indicated by the ensuing FGF2-dependent hyperphosphorylation of the downstream target Rb. In untreated mature OLs that do not incorporate BrdU, the levels of p21Cip1 are low, and the level of the inhibitor p18INK is high. Furthermore, p18INK sequesters CDK2 into binary complexes, precluding the formation of p21Cip1/cyclin D1/CDK4 ternary complexes in these cells. Therefore, we propose that p21Cip1 is acting as a positive regulator, rather than an inhibitor, of cell cycle entry by favoring the assembly of active cyclin D1/CDK4 complexes. PMID- 15789404 TI - Empirical lipid propensities of amino acid residues in multispan alpha helical membrane proteins. AB - Characterizing the interactions between amino acid residues and lipid molecules is important for understanding the assembly of transmembrane helices and for studying membrane protein folding. In this study we develop TMLIP (TransMembrane helix-LIPid), an empirically derived propensity of individual residue types to face lipid membrane based on statistical analysis of high-resolution structures of membrane proteins. Lipid accessibilities of amino acid residues within the transmembrane (TM) region of 29 structures of helical membrane proteins are studied with a spherical probe of radius of 1.9 A. Our results show that there are characteristic preferences for residues to face the headgroup region and the hydrocarbon core region of lipid membrane. Amino acid residues Lys, Arg, Trp, Phe, and Leu are often found exposed at the headgroup regions of the membrane, where they have high propensity to face phospholipid headgroups and glycerol backbones. In the hydrocarbon core region, the strongest preference for interacting with lipids is observed for Ile, Leu, Phe and Val. Small and polar amino acid residues are usually buried inside helical bundles and are strongly lipophobic. There is a strong correlation between various hydrophobicity scales and the propensity of a given residue to face the lipids in the hydrocarbon region of the bilayer. Our data suggest a possibly significant contribution of the lipophobic effect to the folding of membrane proteins. This study shows that membrane proteins have exceedingly apolar exteriors rather than highly polar interiors. Prediction of lipid-facing surfaces of boundary helices using TMLIP1 results in a 54% accuracy, which is significantly better than random (25% accuracy). We also compare performance of TMLIP with another lipid propensity scale, kPROT, and with several hydrophobicity scales using hydrophobic moment analysis. PMID- 15789405 TI - Structural analysis of interactions for complex formation between Ferredoxin NADP+ reductase and its protein partners. AB - The three-dimensional structures of K72E, K75R, K75S, K75Q, and K75E Anabaena Ferredoxin-NADP+ reductase (FNR) mutants have been solved, and particular structural details of these mutants have been used to assess the role played by residues 72 and 75 in optimal complex formation and electron transfer (ET) between FNR and its protein redox partners Ferredoxin (Fd) and Flavodoxin (Fld). Additionally, because there is no structural information available on the interaction between FNR and Fld, a model for the FNR:Fld complex has also been produced based on the previously reported crystal structures and on that of the rat Cytochrome P450 reductase (CPR), onto which FNR and Fld have been structurally aligned, and those reported for the Anabaena and maize FNR:Fd complexes. The model suggests putative electrostatic and hydrophobic interactions between residues on the FNR and Fld surfaces at the complex interface and provides an adequate orientation and distance between the FAD and FMN redox centers for efficient ET without the presence of any other molecule as electron carrier. Thus, the models now available for the FNR:Fd and FNR:Fld interactions and the structures presented here for the mutants at K72 and K75 in Anabaena FNR have been evaluated in light of previous biochemical data. These structures confirm the key participation of residue K75 and K72 in complex formation with both Fd and Fld. The drastic effect in FNR activity produced by replacement of K75 by Glu in the K75E FNR variant is explained not only by the observed changes in the charge distribution on the surface of the K75E FNR mutant, but also by the formation of a salt bridge interaction between E75 and K72 that simultaneously "neutralizes" two essential positive charged side chains for Fld/Fd recognition. PMID- 15789406 TI - Comparison of cell-based and cell-free protocols for producing target proteins from the Arabidopsis thaliana genome for structural studies. AB - We describe a comparative study of protein production from 96 Arabidopsis thaliana open reading frames (ORFs) by cell-based and cell-free protocols. Each target was carried through four pipeline protocols used by the Center for Eukaryotic Structural Genomics (CESG), one for the production of unlabeled protein to be used in crystallization trials and three for the production of 15N labeled proteins to be analyzed by 1H-15N NMR correlation spectroscopy. Two of the protocols involved Escherichia coli cell-based and two involved wheat germ cell-free technology. The progress of each target through each of the protocols was followed with all failures and successes noted. Failures were of the following types: ORF not cloned, protein not expressed, low protein yield, no cleavage of fusion protein, insoluble protein, protein not purified, NMR sample too dilute. Those targets that reached the goal of analysis by 1H-15N NMR correlation spectroscopy were scored as HSQC+ (protein folded and suitable for NMR structural analysis), HSQC+/- (protein partially disordered or not in a single stable conformational state), HSQC- (protein unfolded, misfolded, or aggregated and thus unsuitable for NMR structural analysis). Targets were also scored as X- for failing to crystallize and X+ for successful crystallization. The results constitute a rich database for understanding differences between targets and protocols. In general, the wheat germ cell-free platform offers the advantage of greater genome coverage for NMR-based structural proteomics whereas the E. coli platform when successful yields more protein, as currently needed for crystallization trials for X-ray structure determination. PMID- 15789407 TI - Proliferation of cerebellar neurons induced by astrocytes treated with thyroid hormone is mediated by a cooperation between cell contact and soluble factors and involves the epidermal growth factor-protein kinase a pathway. AB - Cerebellar development is fully dependent on thyroid hormone (T3) levels. We have previously demonstrated a glia-mediated effect of T3 on cerebellar neurons. We have reported that cerebellar astrocytes treated with thyroid hormone secrete epidermal growth factor (EGF), which directly induces neuronal proliferation and, indirectly, by increasing synthesis of extracellular matrix proteins, induces neurite outgrowth in vitro. Here, by using a neuron-astrocyte coculture model, we investigated the involvement of cell contact on neuronal proliferation. Culturing of cerebellar neurons on T3-treated astrocyte carpets or conditioned medium derived from them (T3CM) yielded similar results, revealed by a 60% increase in cell population. However, the absolute number of neurons in coculture assays was greatly enhanced in comparison with that in CM assays (3.5-4-fold). Bromodeoxyuridine (BrdU) incorporation assays revealed that such an increase was due mainly to proliferation of precursors cells. BrdU incorporation was three times higher in cell carpet (31%) than in CM (13%). Treatment of astrocytes by T3 increased neuronal proliferation either by T3CM (2.5 times) or by contact with T3 treated astrocytes (1.5 times). Neuronal death was not affected by T3 treatment of astrocytes as revealed by either trypan blue viability assays or active caspase-3 labeling. Treatment of astrocytes by EGF mimicked T3 effects on neuronal proliferation. Addition of the EGF receptor tyrosine kinase inhibitor genistein and the protein kinase A (PKA) inhibitor KT5720 to cocultutres and T3CM completely reversed neuronal proliferation. Our results implicate EGF and the PKA pathway in the proliferation induced by T3-treated astrocytes. Furthermore, the fact that cocultures potentiate the effect of T3 on neuronal proliferation suggests that neuron-astrocyte contact may cooperate with astrocyte soluble factors to enhance neuronal population. Our data reveal an important role of astrocytes as mediators of T3-induced cerebellar development and partially elucidate the role of cell contact and soluble factors on this process. PMID- 15789408 TI - Identification of a hypaxial somite enhancer element regulating Pax3 expression in migrating myoblasts and characterization of hypaxial muscle Cre transgenic mice. AB - Pax3 encodes a transcription factor that functions in the embryonic central nervous system, neural crest, and somitic mesoderm. Prior studies suggest that distinct regulatory elements regulate temporal and spatial expression of Pax3 in neural crest and mesoderm. Here, we describe a discrete enhancer element, conserved between mouse and human genomes, that directs Pax3 expression in the ventral-lateral lip of interlimb somites. These regions give rise to hypaxial musculature including limb, ventral body wall, diaphragm, and tongue muscles. Transgenic mice harboring the hypaxial muscle enhancer driving lacZ expression initiate beta-galactosidase expression at E10.0, significantly later than endogenous Pax3 expression in presomitic and segmented mesoderm. Initiation of transgene expression is not dependent on Pax3 itself, since expression is detectable in homozygous Splotch embryos. Transgenic mice expressing Cre recombinase in hypaxial myoblasts were generated and characterized. These results suggest that Pax3 is differentially regulated within the somite in both spatial and temporal domains. Hypaxial muscle Cre mice will allow for specific manipulation of gene expression in this subset of developing skeletal muscle. PMID- 15789409 TI - Prediction of fetal anemia in pregnancies with red-cell alloimmunization: comparison of middle cerebral artery peak systolic velocity and amniotic fluid OD450. AB - OBJECTIVE: To compare the accuracy of Doppler velocimetry (middle cerebral artery peak systolic velocity, MCA-PSV) and amniocentesis (amniotic fluid delta optical density 450 (OD450)) for the detection of fetal anemia against the gold standard of fetal blood sampling (FBS). METHODS: Thirty-eight pregnancies were identified to be at risk of fetal anemia from immune causes between January 2000 and May 2002. In a cross-sectional diagnostic accuracy study, MCA-PSV and amniotic fluid delta OD450 values were plotted on reference charts and compared to an FBS obtained within the subsequent 7 days. Receiver-operating characteristics (ROC) curves were used and the area under the curve (AUC) calculated to compare the overall accuracy of the two tests. Sensitivity, specificity and likelihood ratios for positive (LR+) and negative (LR-) test results were generated for specific thresholds of MCA-PSV and delta OD450. RESULTS: For MCA-PSV (n = 38), the AUC was 0.71 (95% CI 0.57-0.85) and for amniotic fluid delta OD450 (n = 22) it was 0.68 (95% CI 0.49-0.87) compared with FBS within 7 days. Sensitivity, specificity and LR+, LR- for MCA-PSV were 64%, 81%, 3.4 and 0.5, respectively, and 53%, 71%, 1.9 and 0.7 for amniotic fluid OD450, respectively. CONCLUSION: MCA-PSV and OD450 have similar test accuracy in detecting fetal anemia. MCA-PSV is non-invasive and therefore presents no risk of miscarriage or preterm labor and thus is a preferable method of screening for fetal anemia. PMID- 15789410 TI - Fgf15 is required for proper morphogenesis of the mouse cardiac outflow tract. AB - Evidence in animal models indicates that signaling networks functioning in the developing pharyngeal arches regulate stereotyped processes critical for proper development of the aortic arch and cardiac outflow tract. Here, we describe the phenotype of mice lacking fibroblast growth factor 15 (Fgf15), which encodes a secreted signaling molecule expressed within the developing pharyngeal arches. Homozygous Fgf15 mutants present heart defects consistent with malalignment of the aorta and pulmonary trunk. These defects correlate with early morphological defects of the outflow tract due to aberrant behavior of the cardiac neural crest. We demonstrate that Fgf15 expression within the pharyngeal arches is unaltered by a loss of Tbx1, a key regulator of pharyngeal arch development implicated in DiGeorge syndrome. In addition, Fgf15 and Tbx1 do not interact genetically, suggesting that Fgf15 operates through a pathway independent of Tbx1. These studies reveal a novel role of Fgf15 during development of the cardiac outflow tract. PMID- 15789411 TI - Effect of argon-plasma treatment on proliferation of human-skin-derived fibroblast on chitosan membrane in vitro. AB - Chitosan is not only a nontoxic, biocompatible, and biodegradable polymer, but has also a chemical structure similar to glycosaminoglycans (GAGs), which promote scarless wound healing of skin. In this study, chitosan membranes were treated with argon plasma to improve their surface hydrophilicity. The results showed that the water contact angles of these surface-treated membranes were significantly reduced from 60.76 to 11.57 degrees . The total surface energy was increased from 41.06 to 67.31 mJ/m(2), with 60-86.95% improvement in the gamma negative component and a 20% difference in the nonpolar component. Argon-plasma treated chitosan membranes exhibited excellent attachment, migration, and proliferation of the human-skin-derived fibroblasts (hSFs) compared to the untreated ones. It was found that the duration of argon-plasma treatment influenced the cell proliferation, and the optical densities in MTT assay were enhanced. Argon-plasma treatment improved the surface hydrophilicity of chitosan membranes and promoted the attachment and proliferation of hSFs. PMID- 15789412 TI - Frequency-dependent recruitment of inhibition mediated by stellate cells in the rat cerebellar cortex. AB - In the cerebellum, dendritic inhibition of Purkinje cells (PCs) is mediated by stellate cells (SCs). These inhibitory interneurons are critically involved in the cerebellar network; they control the timing and firing frequency of PCs, the only output cells of the cerebellar cortex. However, the underlying properties of parallel fiber (PF) to SC excitatory synapses have not been fully determined. To characterize the conditions favoring the recruitment of SCs in the cerebellum, we analyzed evoked and spontaneous excitatory postsynaptic currents (EPSCs) recorded from SCs of rat cerebellar slices. We found that SC EPSCs evoked with single suprathreshold-intensity stimulations were mostly unitary, with a large amplitude and variable latencies, and failed with a high rate. Increasing the frequency of stimulation above 60 Hz significantly reduced failures, whereas mean SC EPSC amplitude was increased by less than 20%. Decreasing failures at PF-SC synapses experimentally enhanced the number of asynchronous SC EPSCs per stimulation but, again, moderately increased the mean SC EPSC amplitude. Finally, brief presynaptic bursts transiently depressed synaptic transmission. This depression resulted from the release of endocannabinoids and might act as a negative feedback mechanism. Thus, we conclude that SC EPSCs evoked with single suprathreshold-intensity stimulations are mostly unitary and that PF-SC synapse efficacy is highly regulated by the presynaptic temporal pattern of activity and the frequency of afferent inputs. Such synaptic properties may control the responsiveness of SC synapses to the frequency of PF stimulations, which may control the spatial extent and duration of the recruitment of inhibition in the cerebellar cortex. PMID- 15789413 TI - Null and conditional semaphorin 3B alleles using a flexible puroDeltatk loxP/FRT vector. AB - In neural development, Semaphorin 3B (SEMA3B) is thought to play a role in guiding axons by repulsion. In nonneuronal tissue, SEMA3B has been postulated to be a tumor suppressor gene of lung and breast cancer. Much of the understanding of the function of members of the SEMA3 family has come from targeted deletion of these genes in mice (Sema3A, Sema3C, and Sema3F). Thus, targeted deletion of Sema3B in mice would prove invaluable in dissecting out its functions. To allow for maximum gene-targeting flexibility, we developed a generic targeting vector, pFlexible, containing the positive/negative selectable marker puroDeltatk and loxP and FRT recombination sites, and used it to target Sema3B in ES cells. Flpe- and Cre-mediated recombination in vitro generated ES cell lines that contained a conditional or null Sema3B allele, respectively, which were established as homozygous alleles in mice. Analysis of Sema3B null mice showed they were viable, fertile, and displayed no overt pathological abnormalities, suggesting an inherent correction mechanism or level of redundancy between the class 3 semaphorins. This targeting vector system has broad applicability in any knockout experiment and provides a flexible approach for the generation of modified alleles in mice. PMID- 15789415 TI - Cell-free fetal DNA in maternal plasma: an important advance to link fetal genetics to obstetric ultrasound. PMID- 15789414 TI - Prenatal diagnosis of a right ventricular diverticulum complicated by pericardial effusion in the first trimester. AB - A ventricular diverticulum associated with a large pericardial effusion was diagnosed at 13 weeks of gestation. The pericardial effusion resolved spontaneously by 20 weeks and the diverticular size remained the same during pregnancy. In the postnatal period the neonate underwent surgical correction of both the diverticulum and associated ventricular and atrial septal defects. Our case indicates that congenital ventricular diverticulum may be associated with a good perinatal outcome. PMID- 15789417 TI - Dissection of the GTPase mechanism of Ras protein by MD analysis of Ras mutants. AB - Controlling the hydrolysis rate of GTP bound to the p21ras protein is crucial for the delicate timing of many biological processes. A few mechanisms were suggested for the hydrolysis of GTP. To gain more insight into the individual elementary events of GTP hydrolysis, we carried out molecular dynamic analysis of wild-type p21ras and some of its mutants. It was recently shown that Ras-related proteins and mutants generally follow a linear free energy relationship (LFER) relating the rate of reaction to the pK(a) of the gamma-phosphate group of the bound GTP, indicating that proton transfer from the attacking water to the GTP is the first elementary event in the GTPase mechanism. However, some exceptions were observed. Thus, the Gly12 --> Aspartic p21ras (G12D) mutant had a very low GTPase activity although its pK(a) was very close to that of the wild-type ras. Here we compared the molecular dynamics (MD) of wild-type Ras and G12D, showing that in the mutant the catalytic water molecule is displaced to a position where proton transfer to GTP is unfavorable. These results suggest that the mechanism of GTPase is indeed composed of an initial proton abstraction from water by the GTP, followed by a nucleophilic attack of the hydroxide ion on the gamma-phosphorus of GTP. PMID- 15789416 TI - Sequence-structure-function relationships of a tRNA (m7G46) methyltransferase studied by homology modeling and site-directed mutagenesis. AB - The Escherichia coli TrmB protein and its Saccharomyces cerevisiae ortholog Trm8p catalyze the S-adenosyl-L-methionine-dependent formation of 7-methylguanosine at position 46 (m7G46) in tRNA. To learn more about the sequence-structure-function relationships of these enzymes we carried out a thorough bioinformatics analysis of the tRNA:m7G methyltransferase (MTase) family to predict sequence regions and individual amino acid residues that may be important for the interactions between the MTase and the tRNA substrate, in particular the target guanosine 46. We used site-directed mutagenesis to construct a series of alanine substitutions and tested the activity of the mutants to elucidate the catalytic and tRNA recognition mechanism of TrmB. The functional analysis of the mutants, together with the homology model of the TrmB structure and the results of the phylogenetic analysis, revealed the crucial residues for the formation of the substrate binding site and the catalytic center in tRNA:m7G MTases. PMID- 15789418 TI - Identification of domains and domain interface residues in multidomain proteins from graph spectral method. AB - We present a novel method for the identification of structural domains and domain interface residues in proteins by graph spectral method. This method converts the three-dimensional structure of the protein into a graph by using atomic coordinates from the PDB file. Domain definitions are obtained by constructing either a protein backbone graph or a protein side-chain graph. The graph is constructed based on the interactions between amino acid residues in the three dimensional structure of the proteins. The spectral parameters of such a graph contain information regarding the domains and subdomains in the protein structure. This is based on the fact that the interactions among amino acids are higher within a domain than across domains. This is evident in the spectra of the protein backbone and the side-chain graphs, thus differentiating the structural domains from one another. Further, residues that occur at the interface of two domains can also be easily identified from the spectra. This method is simple, elegant, and robust. Moreover, a single numeric computation yields both the domain definitions and the interface residues. PMID- 15789420 TI - Gonadal development and sex differentiation in the cichlid fish Cichlasoma dimerus (Teleostei, Perciformes): a light- and electron-microscopic study. AB - Although the overall pattern and timing of gonadal sex differentiation have been established in a considerable number of teleosts, the ultrastructure of early stages of gonadal development is not well documented. In this study, gonads from larval and juvenile stages of laboratory-reared Cichlasoma dimerus were examined at the light-microscopic and ultrastructural levels. This freshwater species adapts easily to captivity and spawns with high frequency during 8 months of the year, providing an appropriate model for developmental studies. Larvae and juveniles were kept at a water temperature of 26.5 +/- 1 degrees C and a 12:12 hour photoperiod. Gonadal development was documented from 14-100 days postfertilization, covering the period of histologically discernible sex differentiation. Gonadal tissue was processed according to standard techniques for light and electron microscopy. C. dimerus, a perciform teleost, is classified as a differentiated gonochorist, in which an indifferent gonad develops directly into a testis or ovary. On day 14, the gonadal primordium consists of a few germ cells surrounded by enveloping somatic cells. Ovarian differentiation precedes testicular differentiation, as usual in teleost fishes. The earliest signs of differentiation, detected from day 42 onward, include the onset of meiotic activity in newly formed oocytes, which is soon accompanied by increased oogonial mitotic proliferation and the somatic reorganization of the presumptive ovary. The ovarian cavity is completely formed by day 65. Numerous follicles containing perinucleolar oocytes are observed by day 100. In contrast, signs of morphological differentiation in the presumptive testis are not observed until day 72. By day 100, the unrestricted lobular organization of the testis is evident. The latest stage of spermatogenesis observed by this time of testicular development is spermatocyte II. PMID- 15789419 TI - Recombineered Xenopus tropicalis BAC expresses a GFP reporter under the control of Arx transcriptional regulatory elements in transgenic Xenopus laevis embryos. AB - The aristaless-related homeobox (Arx) gene is expressed in a dynamic pattern in the developing vertebrate forebrain. We identified a bacterial artificial chromosome (BAC) containing the Xenopus tropicalis Arx gene and replaced a portion of the first coding exon with a green fluorescent protein (GFP) expression cassette by homologous recombination in bacteria (recombineering). Transgenic X. laevis embryos obtained by microinjecting the modified BAC expressed GFP in the developing forebrain in a pattern identical to that of the endogenous Arx gene. Thus, this BAC contains transcriptional regulatory elements necessary for regulating proper expression in transgenic frogs. This work demonstrates that transgenesis using recombineered BACs is a viable technique in Xenopus. PMID- 15789421 TI - Functional morphology of the cephalopod buccal mass: a novel joint type. AB - The arrangement of the musculature and connective tissues of the buccal mass of the coleoid cephalopods Octopus bimaculoides, Sepia officinalis, and Loliguncula brevis was examined using dissection and histology. Serial sections in three mutually perpendicular planes were used to identify the muscles and connective tissues responsible for beak movements and stability and to describe their morphology and fiber trajectories. Four major beak muscles were identified: the anterior, posterior, superior, and lateral mandibular muscles. The anterior, posterior, and superior mandibular muscles connect the upper beak and the lower beak. Although the lateral mandibular muscles originate on the upper beak, they do not connect to the lower beak and instead insert on a connective tissue sheath surrounding the buccal mass. Examination of the fibers of the lateral mandibular muscles reveals that they have the organization of a muscular hydrostat, with muscle fibers oriented in three mutually perpendicular orientations. Although the beaks are capable of complex opening, closing, and shearing movements, they do not contact one another and are instead connected only by the musculature of the buccal mass. Based on the morphological analysis and observations of freshly dissected beaks undergoing the stereotyped bite cycle, the functional role of the beak muscles is hypothesized. The anterior and superior mandibular muscles are likely responsible for beak closing and shearing movements. The posterior mandibular muscle is likely also involved in beak closing, but may act synergistically with the lateral mandibular muscles to open the beaks. The lateral mandibular muscles may use a muscular-hydrostatic mechanism to control the location of the pivot between the beaks and to generate the force required for beak opening. The lack of contact between the beaks and the morphology of the lateral mandibular muscles suggests that the buccal mass of coleoid cephalopods may represent a previously unexamined flexible joint mechanism. The term "muscle articulation" is proposed here to denote the importance of the musculature in the function of such a joint. PMID- 15789422 TI - Analysis of mesenchymal stem cells grown on a three-dimensional HYAFF 11-based prototype ligament scaffold. AB - Ligaments are complex structures that maintain the mechanical stability of the joint. Healing of injured ligaments involves the interactions of different cell types, local cellular environment, and the use of devices. To gain new information on the complex interactions between mesenchymal stem cells (MSCs) and a specific hyaluronan-based prototype scaffold (HYAFF, useful for ligament tissue engineering, short time-course experiments were performed to analyze the proliferation, vitality, and phenotype of MSCs grown on the scaffold. MSC proliferation was analyzed using the MTT test, during the early time points (2, 4, 6, days). Viability was assessed using calcein/acetyloxymethylester immunofluorescence dye and confocal microscopy analysis. Hyaluronic acid receptor (CD44), typical matrix ligament proteins (collagen type I, type III, laminin, fibronectin, actin), and chondrogenic/osteogenic markers (collagen type II and bone sialoprotein) were evaluated by immunohistochemistry. Our data demonstrated that MSC growth and viability were cell density-dependent. MSCs completely wrapped the fibers of the scaffold, expressed CD44, collagen type I, type III, laminin, fibronectin, and actin, and were negative to collagen type II and bone sialoprotein. These data demonstrate that MSCs survive well in the hyaluronan based prototype ligament scaffold, as assessed after 2 days from seeding, and express CD44, a receptor important for scaffold interaction, and proteins responsible for the functional characteristics of the ligaments. PMID- 15789423 TI - High-efficiency somatic mutagenesis in smooth muscle cells and cardiac myocytes in SM22alpha-Cre transgenic mice. AB - The cytoskeletal protein SM22alpha is expressed in visceral and vascular smooth muscle cells (SMCs), in cardiac myocytes, and in the myotomal components of the somites during murine embryonic development. In this report, we describe the generation and characterization of transgenic mice expressing Cre-recombinase under the transcriptional control of the -2.8-kb SM22alpha promoter. Following interbreeding with the R26R reporter strain, Cre-dependent beta-galactosidase expression was observed as early as embryonic day 9.5 in SMCs of the developing vasculature, in cardiac myocytes, but not in the somites. In adult mice, Cre mediated recombination was observed in vascular SMCs throughout the venous and arterial systems, in visceral SMCs in multiple organs, and in cardiac, but not skeletal muscle. Importantly, Cre-mediated recombination was present in nearly 100% of arterial SMCs, including in the aorta. These mice are thus an important new tool for performing in vivo loss-of-function studies of genes expressed in vascular SMCs. PMID- 15789424 TI - Conditional alleles for activation and inactivation of the mouse Rx homeobox gene. AB - The Rx homeobox gene is a transcriptional regulator indispensable for development of the eye and ventral forebrain. Rx-null homozygotes lack optic pits, which are the earliest ocular structures. To study the roles Rx may play at various stages of eye and brain development, we generated an allelic series at the Rx locus. The targeted allele, Rx(neo), is a severely hypomorphic or null allele. This Rx(neo) allele is converted via FLP-mediated recombination to the Rx(flox) allele, which is phenotypically identical to the wildtype allele. Cre-mediated conversion of Rx(flox) generates the RxDelta2 allele, which, when homozygous, results in an Rx null phenotype that includes perinatal lethality, anophthalmia, and anterior neural and craniofacial defects. Mice carrying these alleles allow both Cre mediated inactivation and FLP-mediated activation of Rx gene activity on a conditional basis and will be useful in examining Rx function at different developmental stages and in distinct tissue environments. PMID- 15789425 TI - Temporally controlled targeted somatic mutagenesis in skeletal muscles of the mouse. AB - To generate temporally controlled targeted somatic mutations selectively and efficiently in skeletal muscles, we established a transgenic HSA-Cre-ER(T2) mouse line in which the expression of the tamoxifen-dependent Cre-ER(T2) recombinase is under the control of a large genomic DNA segment of the human skeletal muscle alpha-actin gene, contained in a P1-derived artificial chromosome. In this transgenic line Cre-ER(T2) is selectively expressed in skeletal muscles, and Cre ER(T2)-mediated alteration of LoxP flanked (floxed) target genes is skeletal muscle-specific and strictly tamoxifen-dependent. HSA-Cre-ER(T2) mice should be of great value to analyze gene function in skeletal muscles, and to establish animal models of human skeletal muscle disorders. PMID- 15789426 TI - Temporally regulated expression of Cre recombinase in neural stem cells. AB - The use of mouse gene targeting to study molecules important in neural development is oftentimes impaired by early embryonic lethality. In order to address later roles for such molecules, specifically in neural stem cells, we generated transgenic mice that express both the tetracycline-inducible molecule rtTA-M2 and GFP under the control of the neural precursor specific form of nestin. Developmental analysis of these mice demonstrates that GFP expression is exclusive to the neural tube. Adult expression of GFP is seen only in known areas of adult neurogenesis, namely, the subventricular zone and the dentate gyrus. When crossed with a second transgenic mouse (TetOp-Cre) that expresses the Cre recombinase under the control of the tetracycline responsive promotor, we demonstrate temporal induction of Cre in bigenic animals exposed to doxycycline. We further demonstrate the feasibility of this approach by using the ROSA-26 reporter mouse to mediate recombination in neural precursor cells. PMID- 15789427 TI - Whole-mount analysis reveals normal numbers of dopaminergic neurons following misexpression of alpha-Synuclein in Drosophila. AB - Previously published reports have suggested that misexpression of alpha-Synuclein in the Drosophila central nervous system causes neurodegeneration and progressive age-dependent locomotor dysfunction similar to pathologic and clinical manifestations of Parkinson's disease. The number of dopaminergic (DA) neurons in these studies was assessed using immunohistochemistry with an anti-tyrosine hydroxylase antibody on sequential paraffin sections of fly brains. In contrast, we do not observe any DA cell loss in alpha-Synuclein expressing fly brains when using whole-mount immunohistochemistry as an assay. Our results suggest that the DA cell loss observed with misexpression of alpha-Synuclein is not fully penetrant under a variety of experimental conditions and that this may complicate interpretation of such experiments. PMID- 15789428 TI - Quantum study of mutational effect in binding of efavirenz to HIV-1 RT. AB - Full quantum mechanical computational study has been carried out to study binding of efavirenz (EFZ), a second generation FDA approved nonnucleoside inhibitor, to HIV-1 reverse transcriptase (RT) and its K103N and Y181C mutants using the MFCC (molecular fractionation with conjugate caps) method. The binding interaction energies between EFZ and each protein fragment are calculated using a combination of HF/3-21G, B3LYP/6-31G* and MP2/6-31G* ab initio levels. The present computation shows that Efavirenz binds to HIV-1 RT predominantly through strong electrostatic interaction with the Lys101 residue. The small loss of binding to K103N mutant by Efavirenz can be attributed to a slightly weakened attractive interaction between the drug and Lys101 due to a conformational change of mutation. The small loss of binding to Y181C mutant by efavirenz can be attributed to the Glu698 residue moving closer to EFZ due to conformational change, which results in an increase of repulsive energy relative to the wild type (WT). The binding of efavirenz-derived DPC961 to HIV-1 RT is enhanced by an additional attractive interaction to residue Hid235 and reduced repulsion to Glu698, resulting in an increase of binding energy by about 4 kcal/mol. PMID- 15789429 TI - Genetic expression profile of olfactory ensheathing cells is distinct from that of Schwann cells and astrocytes. AB - Olfactory ensheathing cells (OECs) accompany the axons of olfactory receptor neurons, which regenerate throughout life, from the olfactory mucosa into the olfactory bulb. OECs have shown widely varying efficacy in repairing the injured nervous system. Analysis of the transcriptome of OECs will help in understanding their biology and will provide tools for investigating the mechanisms of their efficacy and interactions with host tissues in lesion models. In this study, we compared the transcriptional profile of cultured OECs with that of Schwann cells (SCs) and astrocytes (ACs), two glial cell types to which OECs have similarities. Two biological replicates of RNA from cultured OECs, SCs, and ACs were hybridized to long oligo rat 5K arrays against a common reference pool of RNA (50% cultured fibroblast RNA and 50% neonatal rat brain RNA). Transcriptional profiles were analyzed by hierarchical clustering, Principal Components Analysis, and the Venn diagram. The three glial cell types had similarly increased or decreased expression of numerous transcripts compared with the reference. However, OECs were distinguishable from both SCs and ACs by a modest number of transcripts, which were significantly enriched or depleted. Furthermore, OECs and SCs were more closely related to each other than to ACs. Expression of selected transcripts not previously characterized in OECs, such as Lyz, Timp2, Gro1 (Cxcl1), Ccl2 (MCP1), Ctgf, and Cebpb, was validated by real-time reverse transcription-polymerase chain reaction (RT-PCR); immunohistochemistry in cultured OECs, SCs, and ACs, and adult tissues was performed to demonstrate their expression at the protein level. PMID- 15789430 TI - Distribution of Aquaporin 4 in rodent spinal cord: relationship with astrocyte markers and chondroitin sulfate proteoglycans. AB - Water balance between cells and extracellular compartments is essential for proper functioning of the central nervous system, as demonstrated by its perturbations in pathological conditions. Aquaporin 4 (AQP4) is the predominant water channel in brain and spinal cord, where it is present mainly on astrocytic endfeet contacting vessels. A role in water homeostasis control has been proposed also for the extracellular matrix, that in brain consists mainly of chondroitin sulfate proteoglycans (CSPGs). Using cytochemical and immunocytochemical techniques, we investigated their distribution in rodent spinal cord, to better understand the role of these two classes of molecules. The results show that in spinal gray matter AQP4 labeling is intense in all perivascular profiles and (1) displays a marked dorsoventral gradient in the neuropil; and (2) coexists extensively with glial glutamate transporter-1 (GLT-1) but scarcely with glial fibrillary acidic protein (GFAP). In white matter the overlap between AQP4, GLT 1, and GFAP is almost complete. Ultrastructural examination shows that AQP4 labeled astrocytic processes surround blood vessels, neuronal perikarya and processes, and both asymmetric and symmetric synapses, indicating that the protein may be involved in the regulation of water fluxes around both inhibitory and excitatory synapses. CSPGs, visualized by labeling with Wisteria floribunda agglutinin, show a distribution complementary to that of AQP4, being absent or weekly expressed in AQP4-enriched areas. These findings suggest that different mechanisms may contribute to the regulation of water homeostasis in different spinal cord regions. PMID- 15789431 TI - Lipopolysaccharide increases microglial GLT-1 expression and glutamate uptake capacity in vitro by a mechanism dependent on TNF-alpha. AB - This study investigates the effect of microglial activation on microglial glutamate transporters in vitro. Stimuli known to activate microglia and/or to be associated with pathological conditions with an impaired astroglial glutamate uptake were compared. Morphological changes and marked increases in ED1 antigen expression were found after 8-h incubation of rat microglia in 56 mM KCl, 1 ng/ml lipopolysaccharide (LPS), and 100 microM glutamate, as well as in acidic and basic conditions, showing that the cells were activated. Of the stimuli used, only LPS induced a significant release of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), and was the only stimulus that increased microglial GLT-1 expression and glutamate uptake capacity after 12-h incubation. This effect was probably mediated by TNF-alpha, since this cytokine mimicked the effect of LPS. Furthermore, the effect of LPS was blocked by thalidomide, an inhibitor of TNF-alpha synthesis. Additionally, neutralizing antibodies against TNF-alpha also blocked the increase, indicating TNF-alpha as an inducer of GLT-1 expression in microglia. It was also found that preincubation with glutamate dose-dependently inhibited the microglial glutamate uptake. This could suggest different physiological functions for microglial and astroglial glutamate uptake and might indicate a reciprocal control of GLT-1 expression between microglia and astrocytes. PMID- 15789432 TI - Activity-dependent change in morphology of the glial tubular lattice of the crayfish medial giant nerve fiber. AB - An evaluation of electron micrographs of stimulated nerve fibers used to investigate the effect of action potential generation on the structure-function relationship between axons and its associated glial cells revealed that what was at first thought to be stimulation-induced damage to the glia was, in fact, limited to volume expansion and disaggregation of the glial tubular lattice. All other structures appeared well preserved and otherwise normal. Using a 4-point subjective scale for evaluation by two investigators, 50-Hz stimulation for 2 min was observed to cause a volume expansion and disaggregation of the tubular lattice. Quantitatively, the internal diameter of the stimulated tubular lattice increased 65% above the unstimulated control (50.96 +/- 2.09 nm and 30.81 +/- 0.87 nm, respectively, P < or = 0.001). Stimulation had its greatest effect on tubular lattice volume and organization in the adaxonal glial layer and a decreasing effect as distance from the giant axon increased. These effects are reversible since the tubular lattice diameter and degree of disaggregation preserved 10 min after the cessation of stimulation were not found to be different from their unstimulated paired controls. Axons injected with TEA, a voltage-gated potassium channel blocker, prevented stimulation-induced volume expansion and disaggregation of tubular lattice structure. These results are consistent with an active uptake of K+ with obligated water or, alternatively, hyperosmotic K+ uptake and a fixation-induced increase in water permeation. Either mechanism of K+ uptake would result in tubular lattice volume expansion and disaggregation and suggests that the tubular lattice serves a larger role than a simple trans-glial diffusion pathway. PMID- 15789433 TI - PPRODO: prediction of protein domain boundaries using neural networks. AB - Successful prediction of protein domain boundaries provides valuable information not only for the computational structure prediction of multidomain proteins but also for the experimental structure determination. Since protein sequences of multiple domains may contain much information regarding evolutionary processes such as gene-exon shuffling, this information can be detected by analyzing the position-specific scoring matrix (PSSM) generated by PSI-BLAST. We have presented a method, PPRODO (Prediction of PROtein DOmain boundaries) that predicts domain boundaries of proteins from sequence information by a neural network. The network is trained and tested using the values obtained from the PSSM generated by PSI BLAST. A 10-fold cross-validation technique is performed to obtain the parameters of neural networks using a nonredundant set of 522 proteins containing 2 contiguous domains. PPRODO provides good and consistent results for the prediction of domain boundaries, with accuracy of about 66% using the +/-20 residue criterion. The PPRODO source code, as well as all data sets used in this work, are available from http://gene.kias.re.kr/ approximately jlee/pprodo/. PMID- 15789434 TI - Addressing the intrinsic disorder bottleneck in structural proteomics. AB - The Center for Eukaryotic Structural Genomics (CESG), as part of the Protein Structure Initiative (PSI), has established a high-throughput structure determination pipeline focused on eukaryotic proteins. NMR spectroscopy is an integral part of this pipeline, both as a method for structure determinations and as a means for screening proteins for stable structure. Because computational approaches have estimated that many eukaryotic proteins are highly disordered, about 1 year into the project, CESG began to use an algorithm (the Predictor of Naturally Disordered Regions, PONDR to avoid proteins that were likely to be disordered. We report a retrospective analysis of the effect of this filtering on the yield of viable structure determination candidates. In addition, we have used our current database of results on 70 protein targets from Arabidopsis thaliana and 1 from Caenorhabditis elegans, which were labeled uniformly with nitrogen-15 and screened for disorder by NMR spectroscopy, to compare the original algorithm with 13 other approaches for predicting disorder from sequence. Our study indicates that the efficiency of structural proteomics of eukaryotes can be improved significantly by removing targets predicted to be disordered by an algorithm chosen to provide optimal performance. PMID- 15789435 TI - Thrombin-activated microglia contribute to death of dopaminergic neurons in rat mesencephalic cultures: dual roles of mitogen-activated protein kinase signaling pathways. AB - This study evaluated the role of thrombin-activated microglia in the neurodegeneration of mesencephalic cultures. Immunocytochemical and biochemical evidence indicated that in co-cultures consisting of rat cortical microglia and mesencephalic neurons, thrombin led to nonselective loss of mesencephalic neurons. Accompanying neurodegeneration, microglial activation was obvious, evidenced by expression of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-1beta, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) and by increasing production of TNF-alpha and nitric oxide (NO). In mesencephalic neurons treated with conditioned media (CM) taken from thrombin-activated microglia, the number of dopaminergic neurons was significantly attenuated. The neurotoxicity of the CM was diminished when it was derived from microglia co treated with thrombin and either an extracellular signal-regulated kinase 1/2 (ERK1/2) pathway inhibitor (PD98059) or a p38-mitogen-activated protein kinase (p38-MAPK) inhibitor (SB203580). Moreover, jun N-terminal kinase (JNK) and p38 MAPK were activated in mesencephalic neurons treated with CM of thrombin activated microglia. Inhibition of JNK and p38-MAPK rescued the dopaminergic neurons. Collectively, these results indicate that thrombin-activated microglia induce neurodegeneration in cultured mesencephalic neurons and that the MAPKs actively participate in both microglial activation and neurodegeneration. The present data carefully suggest that microglial activation triggered by thrombin may be involved in the neuropathological processes of dopaminergic neuronal cell death that occur in Parkinson's disease. PMID- 15789436 TI - Thermodynamic and kinetic characterization of a beta-hairpin peptide in solution: an extended phase space sampling by molecular dynamics simulations in explicit water. AB - The folding of the amyloidogenic H1 peptide MKHMAGAAAAGAVV taken from the syrian hamster prion protein is explored in explicit aqueous solution at 300 K using long time scale all-atom molecular dynamics simulations for a total simulation time of 1.1 mus. The system, initially modeled as an alpha-helix, preferentially adopts a beta-hairpin structure and several unfolding/refolding events are observed, yielding a very short average beta-hairpin folding time of approximately 200 ns. The long time scale accessed by our simulations and the reversibility of the folding allow to properly explore the configurational space of the peptide in solution. The free energy profile, as a function of the principal components (essential eigenvectors) of motion, describing the main conformational transitions, shows the characteristic features of a funneled landscape, with a downhill surface toward the beta-hairpin folded basin. However, the analysis of the peptide thermodynamic stability, reveals that the beta hairpin in solution is rather unstable. These results are in good agreement with several experimental evidences, according to which the isolated H1 peptide adopts very rapidly in water beta-sheet structure, leading to amyloid fibril precipitates [Nguyen et al., Biochemistry 1995;34:4186-4192; Inouye et al., J Struct Biol 1998;122:247-255]. Moreover, in this article we also characterize the diffusion behavior in conformational space, investigating its relations with folding/unfolding conditions. PMID- 15789439 TI - Pathophysiologic mechanisms of persistent pulmonary hypertension of the newborn. AB - Persistent pulmonary hypertension of the newborn (PPHN), among the most rapidly progressive and potentially fatal of vasculopathies, is a disorder of vascular transition from fetal to neonatal circulation, manifesting as hypoxemic respiratory failure. PPHN represents a common pathway of vascular injury activated by numerous perinatal stresses: hypoxia, hypoglycemia, cold stress, sepsis, and direct lung injury. As with other multifactorial diseases, a single inciting event may be augmented by multiple concurrent/subsequent phenomena that result in differing courses of disease progression. I review the various mechanisms of vascular injury involved in neonatal pulmonary hypertension: endothelial dysfunction, inflammation, hypoxia, and mechanical strain, in the context of downstream effects on pulmonary vascular endothelial-myocyte interactions and myocyte phenotypic plasticity. PMID- 15789440 TI - Design of translactam HCMV protease inhibitors as potent antivirals. AB - Human cytomegalovirus (HCMV) is an important pathogen for which there is a significant unmet medical need. New HCMV antivirals, active against novel molecular targets, are undoubtedly needed as the currently available drugs ganciclovir, cidofovir, and foscarnet, which are all viral DNA inhibitors, suffer from limited effectiveness, mainly due to the development of drug resistance, poor bioavailability, and toxicity. One of the newer molecular targets that has been exploited in the search for better drug candidates is HCMV protease. Our deltaAla HCMV protease (wild type variant with the internal cleavage site deleted) was cloned and expressed in E. coli. This viral enzyme was used to develop HCMV protease assays to evaluate potential inhibitors. The chirally pure (SRS)-alpha-methyl pyrrolidine-5,5-trans-lactam template was synthesized, which together with the natural substrate requirements of HCMV protease and detailed SAR, was used to design potent and selective mechanism based inhibitors of HCMV protease. The mechanism of action of these inhibitors of HCMV protease was investigated by ESI/MS, and the X-ray crystal structure of the HCMV protease was used to refine our selective viral enzyme inhibitors to obtain plasma stable antivirals. A novel ELISA antiviral assay was developed which, together with a cytotoxicity assay, enabled us to discover anti-HCMV drug candidates equivalent in potency to ganciclovir that had good pharmacokinetics in the dog and good brain and ocular penetration in the guinea pig. PMID- 15789441 TI - Interstitial lung disease in patients with ataxia-telangiectasia. AB - Ataxia-telangiectasia (A-T) is an autosomal-recessive multiorgan disease characterized by progressive neurologic deterioration in which the most common causes of death are diseases of the respiratory system and cancers. The aim of this retrospective study was to delineate the clinical, radiographic, and pathologic manifestations of the chronic progressive interstitial lung disease seen in patients with A-T. The medical records of 97 patients with A-T and chronic lung disease were reviewed. Interstitial lung disease (ILD) was specifically diagnosed in 25 of 97 patients. None of these patients had evidence of an infectious process preceding the onset of their lung disease, and none had lasting clinical improvement after treatment with antibiotics. Although many medications were used to treat their ILD, only treatment with systemic corticosteroids early in the course of their illness was associated with clinical and radiographic improvement. Nineteen of these 25 patients with ILD died within 24 months of the onset of their ILD, and of 7 patients treated with corticosteroids, 5 are still alive. Recognition of interstitial lung disease in patients with A-T and its early treatment could reduce or eliminate pulmonary disease as a leading cause of death for these patients. PMID- 15789442 TI - Influence of socioeconomic status on lung function and prediction equations in Indian children. AB - The present study was carried out to assess the influence of socioeconomic status on lung functions and to suggest prediction equations for Indian children. For this purpose, 2,616 normal, healthy schoolchildren aged between 5-15 years were recruited. Boys were classified into three groups, i.e., high-income (HIG), middle-income (MIG), and low-income (LIG), while girls were classified into HIG and LIG groups, based on socioeconomic status (SES). Height, weight, chest circumference, body surface area (BSA), fat-free mass (FFM), and body fat were assessed. Forced expiratory volume in 1 sec (FEV1), forced vital capacity (FVC), FEV1/FVC ratio, and peak expiratory flow rate (PEFR) were measured. The results, before and after adjustment of physical characteristics, showed that anthropometry, body composition, and lung functions were significantly higher in HIG compared to MIG and LIG children, while in girls, no differences were observed in physical characteristics after adjustments. Multiple linear regression equations were developed to predict FEV1, FVC, and PEFR, using independent variables like age, height, fat-free mass, and SES. It is opined that these equations could be used as Indian reference equations for healthy children based on the SES. PMID- 15789443 TI - Expression of SRY proteins in both normal and sex-reversed XY fetal mouse gonads. AB - Sry, a single-copy gene on the Y-chromosome, acts dominantly to trigger differentiation of a testis from a gonadal primordium that otherwise develops into an ovary in mammals. Sry encodes a protein with a DNA-binding domain and probably acts as a transcription factor. However, the mode of SRY action in testis determination remains largely unknown. In the present study, we detected the endogenous SRY protein in normal XY fetal mouse gonads by Western blotting and immunohistochemistry. The tissue-specificity and ontogeny of the detected protein were consistent with those of Sry transcripts. Immunofluorescent double labeling revealed that the SRY protein was detected in the Sertoli cell lineage and was swiftly down-regulated concurrently with testis cord organization. Surprisingly, however, the SRY protein was detected in the entire gonad from the onset of its expression, not in parallel to the spatiotemporal pattern of testis cord organization. The SRY protein was also detected in the entire region of all B6.Y(TIR) fetal gonads, which were anticipated to undergo either partial or complete sex reversal. SRY down-regulation was considerably delayed, compared with control B6.XY gonads and was not associated with testis cord organization in B6.Y(TIR) gonads. We conclude that the testis-determining pathway is impaired at the site of SRY action in the B6.Y(TIR) gonad. PMID- 15789444 TI - Homozygous Ft embryos are affected in floor plate maintenance and ventral neural tube patterning. AB - Sonic hedgehog (Shh), produced by the notochord and floor plate cells of the neural tube, plays a critical role in organizing dorsal-ventral patterning in the developing neural tube. We have investigated neural tube development in mouse embryos homozygous for the Fused toes (Ft) mutation, a deletion composed of genes of the Iroquois B (IrxB) cluster and of Fts, Ftm, and Fto. In Ft mutants starting from embryonic day 10.5, the floor plate appeared to degenerate and the notochord failed to undergo ventral displacement from the spinal cord. Consistent with the loss of Shh signalling from the floor plate, V3 neuron generation was reduced in Ft/Ft embryos and the domain of motor neuron generation expanded ventrally at the expense of V2 neurons. These data support the idea that Ft genes play an important role in dorsal-ventral patterning of the neural tube acting to define the extent of motor neuron generation; moreover, the data reveal a previously unanticipated function for Ft genes in the maintenance of the floor plate. PMID- 15789445 TI - Identification of cDNAs associated with late dedifferentiation in adult newt forelimb regeneration. AB - Epimorphic limb regeneration in the adult newt involves the dedifferentiation of differentiated cells to yield a pluripotent blastemal cell. These mesenchymal like cells proliferate and subsequently respond to patterning and differentiation cues to form a new limb. Understanding the dedifferentiation process requires the selective identification of dedifferentiating cells within the heterogeneous population of cells in the regenerate. In this study, representational differences analysis was used to produce an enriched population of dedifferentiation-associated cDNA fragments. Fifty-nine unique cDNA fragments were identified, sequenced, and analyzed using bioinformatics tools and databases. Some of these clones demonstrate significant similarity to known genes in other species. Other clones can be linked by homology to pathways previously implicated in the dedifferentiation process. These data will form the basis for further analyses to elucidate the role of candidate genes in the dedifferentiation process during newt forelimb regeneration. PMID- 15789446 TI - Wnt10a is involved in AER formation during chick limb development. AB - The apical ectodermal ridge (AER) is indispensable for vertebrate limb development and requires Wnt/beta-catenin signaling for induction and maintenance. We report identification and involvement of Wnt10a in AER formation during chick limb development. Chicken Wnt10a has 82% identity with mouse Wnt10a in the amino acid sequence. The Wnt10a gene was expressed broadly in the surface ectoderm from as early as stage 10. By stage 15, the expression was restricted to the surface ectoderm overlying the lateral plate mesoderm. Wnt10a expression became intensified in the presumptive limb ectoderm during AER formation, and subsequently intense expression signals persisted in the AER. Wnt10a misexpression led to ectopic Fgf8 expression in the developing limb ectoderm and induced translocation of beta-catenin in chick embryo fibroblasts. These results suggest that Wnt10a is involved in AER formation in the chick limb bud through the Wnt/beta-catenin signaling pathway. PMID- 15789447 TI - Bayesian modelling of multivariate quantitative traits using seemingly unrelated regressions. AB - We investigate a Bayesian approach to modelling the statistical association between markers at multiple loci and multivariate quantitative traits. In particular, we describe the use of Bayesian Seemingly Unrelated Regressions (SUR) whereby genotypes at the different loci are allowed to have non-simultaneous effects on the phenotypes considered with residuals from each regression assumed correlated. We present results from simulations showing that, under rather general conditions that are likely to hold in real situations, the Bayesian SUR approach has increased probability of selecting the true model compared to univariate analyses. Finally, we apply our methods to data from subjects genotyped for 12 SNPs in the apolipoprotein E (APOE) gene. Phenotypes relate to response to treatment with atorvastatin and include changes in total cholesterol, low-density lipoprotein cholesterol, and triglycerides. Missing genotype data are naturally accommodated in our Bayesian framework by imputing them using a nested haplotype phasing algorithm. PMID- 15789448 TI - [Expression of matrix metalloproteinase-2 in rats with liver fibrosis]. PMID- 15789449 TI - Epidemiological Freudianism. PMID- 15789450 TI - HIV/AIDS: doing what's right. PMID- 15789452 TI - "Breakthrough" in diabetes treatment? PMID- 15789451 TI - Health must come before politics in WHO--Africa's reforms. PMID- 15789453 TI - Can Cyprus overcome its health-care challenges? PMID- 15789454 TI - Guidelines for reporting statistics in journals published by the American Physiological Society. PMID- 15789455 TI - Ablation for atrial fibrillation in congestive heart failure. PMID- 15789456 TI - The "pill-in-the-pocket" approach to atrial fibrillation. PMID- 15789457 TI - The "pill-in-the-pocket" approach to atrial fibrillation. PMID- 15789458 TI - Vascular events after acute infection or vaccination. PMID- 15789459 TI - Acute pericarditis. PMID- 15789460 TI - Acute pericarditis. PMID- 15789461 TI - H1-antihistamines. PMID- 15789462 TI - Nurse migration is only a symptom, not the disease. Interview by Jan Harrington. PMID- 15789463 TI - Breast cancer in Lebanon. Increased age-adjusted incidence rates in younger-aged groups at presentation: implications for screening and for Arab-American ethnic groups. PMID- 15789464 TI - Knowledge, beliefs, and attitudes about environmental diseases among Arab American communities in the Detroit area. PMID- 15789465 TI - Community-based participatory research of environmental factors on Arab Americans in metropolitan Detroit: progress report. PMID- 15789466 TI - Arab-American youth tobacco program. PMID- 15789467 TI - Predictors of tobacco use among Lebanese, Yemeni, and Iraqi adolescents, 14-18 years of age. PMID- 15789469 TI - Federation survey shows opposition to Medicare, Medicaid cuts. PMID- 15789468 TI - A comparison of psychosocial factors and tobacco use among Arab and Arab-American adolescents: preliminary findings. PMID- 15789470 TI - High antiretroviral drug adherence key in effort to avoid drug resistance. New study highlights its importance. AB - A new study confirms the findings of previous research that antiretroviral drug adherence is a strong predictor of whether or not drug resistance occurs in HIV patients. Investigators at the British Columbia Centre for Excellence Research Labs in Vancouver looked for signs of HIV drug resistance among 1,200 HIV patients. PMID- 15789471 TI - HIV clinic improves medication adherence. DOT is latest experimental strategy. AB - HIV clinicians treating patients who are failing their antiretroviral drug regimens face a conundrum: how do you increase adherence to increasingly difficult medication regimens among a population that has developed drug resistance most likely due to poor adherence in the first place? PMID- 15789472 TI - Sex education distorts information on condoms. Key to prevention, condoms, often are ignored. AB - HIV and public health experts say the condom remains the best prevention tool available in the arsenal against HIV infection, yet no one is passing out condoms as readily as they should. Likewise, the trend in the United States appears to be heading in the other direction of further restricting condom distribution among youth. PMID- 15789473 TI - Teen sexual risk behavior news is both good and bad. Experts promote comprehensive education. AB - HIV prevention and sexual education in schools have produced both a good picture and a bad picture, experts say. The good news is more young people are using condoms. The bad news: they are used more in new relationships but no as much in the long term. PMID- 15789474 TI - Africa, India test sites for anti-HIV microbicide. Cellulose sulfate tested in high-risk women. AB - Phase III effectiveness clinical trials have begun for the potential anti-HIV microbicide cellulose sulfate (Ushercell), formerly called C31G, which already has demonstrated safety when used by women. If the trials go well, the product could be ready for approval by the FDA by 2010. PMID- 15789475 TI - Vaccine news: new meningitis vaccine approved for children...and should we be giving all schoolchildren flu vaccine? PMID- 15789476 TI - Fast food really can be unhealthy...and new dietary guidelines. PMID- 15789478 TI - Child safety. Poison prevention awareness. PMID- 15789477 TI - More on color blindness. PMID- 15789479 TI - [Portal and splenic veins thrombosis reveling a miliary tuberculosis of the liver]. AB - The etiologies of the portal vein trombosis are dominated by the neoplasic forms with hepatocellular carcinoma; we report a rare case of portal and splenic veins thrombosis revealing a liver military tuberculosis occuring in a HIV 1 infected patient. A 42 years old senegalese woman with no personal or family history of thrombosis was admitted for abdominal upper right quadran, and epigastric pain, with fever and important weight loss. Ultrasound identified endoluminal echogenic images in the portal and splenic veins. There were no lymph nodes or liver tumor. Evaluations of proteins C and S were normal and there was no anticardiolipin antibody. In searching the aetiology of the thrombosis, a liver biopsy was performed, and showed a miliary tuberculosis. an HIV 1 infection was later on diagnosed. The antituberculosis treatment associated with heparine therapy was successful, the thrombosis resolved entirely. This portal and splenic veins thromboses occuring on a miliary tuberculosis of the liver seems to be an exceptional situation. We did not found in the literature a similar case.lt points out the interset of liver biopsy in searching the aetiology and the early heparine therapy set up PMID- 15789480 TI - [Consensus over terminology and concepts for lower urinary tract function]. PMID- 15789481 TI - End-of-life care. National Institutes of Health statement on the state of the science. PMID- 15789482 TI - Rainbow tags: a visual tag system for recombinant protein expression and purification. AB - Visualization systems for tracking proteins are standard experimental tools in most areas of biological research apart from protein purification. Here, we have sought to plug this gap by producing red and yellow visual tags using the heme binding domain of mosquito cytochrome b5 and the flavin mononucleotide (FMN) binding domain of human P450 reductase. Tests with colorless glutathione-S transferase (GST) show them to be simple and effective tools for visually identifying correctly folded protein and tracking protein molecules through protein expression and purification. Furthermore, the characteristic absorbance signatures of the colored tags can be used to quantify protein concentrations directly, which allows purification to be linked to colorimetric detection. This technology, which we call Rainbow Tagging, facilitates expression and downstream processing of recombinant proteins, paving the way for the development of automated high-throughput protein expression systems. PMID- 15789483 TI - Prevention: the best cure for occupational asthma. PMID- 15789484 TI - [From the model to regulation offer -- political dialogue with reform schools for nursing professions]. PMID- 15789485 TI - The toxicity and bioavailability of atrazine and molinate to Chironomus tepperi larvae in laboratory and river water in the presence and absence of sediment. AB - Acute (10 day) semi-static toxicity tests in which the midge, Chironomus tepperi, were exposed to atrazine and molinate were conducted in laboratory water and in river water, in the absence and presence of sediment. The bioavailability measured as median lethal concentrations (LC50) and 95% fiducial limits (FLs) of atrazine to C. tepperi in laboratory water in the absence and presence of sediment were 16.6 (14.3-19.4) and 21.0 (18.2-24.1) mg l(-1), respectively while the corresponding values in river water were 16.7 (14.7-19.0) and 22.7 (20.3 25.4) mg l(-1), respectively. For molinate, the LC50 and FL values in laboratory water in the absence and presence of sediment were 8.8 (6.8-11.4) and 14.3 (12.4 16.4) mg l(-1), respectively and the corresponding values in river water were 9.3 (7.6-11.3) and 14.5 (12.4-16.9) mg l(-1), respectively. Atrazine has low toxicity (LC50 > 10 mg l(-1)) while molinate has moderate toxicity (1 mg l(-1) < LC50 < 10 mg l(-1)) to C. tepperi. River water did not significantly (P > 0.05) reduce the bioavailability of either chemical to C. tepperi. However, the presence of sediment did significantly (P < 0.05) reduce the bioavailability of both atrazine and molinate to C. tepperi. PMID- 15789486 TI - From the environment to the workplace and back again? Occupational health and safety activism in Ontario, 1970s-2000+. AB - This article addresses the issue of alliances between social movements through an investigation into the possibility of a convergence and/or allliance between two of the most enduring social movements in Ontario over the past three decades: the occupational health and safety (OHS) and the environmental movements. Both movements have not only endured, but been successful movements. Presently, however, each movement is fragmented and becalmed. Recent political developments are leading many OHS/labour movement activists to argue for an alliance with environmental movements. I argue that while there is benefit to both movements in such undertakings, continuing class differences between these two movements militates against any enduring alliance. PMID- 15789487 TI - Between two worlds: Yamanouchi Shigeo and eugenics in early twentieth-century Japan. AB - This paper explores the eugenic through of Yamanouchi Shigeo (1876-1973), who was trained in plant cytology under the tutelage of botanist and eugenicist John Coulter (1851-1928) in the USA, and later become one of the early and important popularizers of eugenic ideas in Japan. His career demonstrates a direct link between Japanese and US eugenics. Despite his academic training and research at various internationally renowned institutions, numerous publications, and longevity, his life has received little scholarly attention. By the early twentieth century, most biologists in Japan, as in the USA, began accepting Mendelian evolutionary theory and rejecting the Lamarckian notion of inheritance of acquire characteristics. However, Yamanouchi Shigeo's eugenic view represents a paradox: he was a mendelian cytologist sympathetic to Lamarckism. Was his 'nurture'-oriented eugenic view unscientific? is that why he was largely ignored in the history of botany in Japan? This study attempts to answer these questions and to analyse the origins and distinct features of Yamanouchi's eugenic ideas by situating Yamanouchi's eugenic through historically and culturally. After examining his scientific papers, popular writings, and documents of various organizations to which he belonged, I argue that Yamanouchi's 'softer' (or less biologically deterministic) perspective may have reflected the Japanese desire to catch up with the dominant 'race' by using eugenics without accepting permanent inferior status. PMID- 15789488 TI - Early laboratories c.1600 - c.1800 and the location of experimental science. AB - Surprisingly little attention has been given hitherto to the definition of the laboratory. A space has to be specially adapted to deserve that title. It would be easy to assume that the two leading experimental sciences, physics and chemistry, have historically depended in a similar way on access to a laboratory. But while chemistry, through its alchemical ancestry with batteries of stills, had many fully fledged laboratories by the seventeenth century, physics was discovering the value of mathematics. Even experimental physics was content to make use of almost any indoor space, if not outdoors, ignoring the possible value of a laboratory. The development of the physics laboratory had to wait until the nineteenth century. PMID- 15789489 TI - Malaria and travel to the Dominican Republic. PMID- 15789490 TI - Buruli ulcer distribution in Benin. PMID- 15789491 TI - Mycobacterium tuberculosis drug resistance, Abkhazia. PMID- 15789492 TI - Endogeneity in logistic regression models. PMID- 15789493 TI - Rectal lymphogranuloma venereum, France. PMID- 15789494 TI - Pertussis in soldiers, Israel. PMID- 15789495 TI - Food safety for first responders. PMID- 15789496 TI - Rickettsiae in ixodid ticks, Sicily. PMID- 15789497 TI - Rats, lice, and Zinsser. PMID- 15789498 TI - Presence of antispasmodic, antidiarrheal, antisecretory, calcium antagonist and acetylcholinesterase inhibitory steroidal alkaloids in Sarcococca saligna. AB - The aim of this investigation was to see if the crude extract of Sarcococca saligna (Ss.Cr) contains chemicals with gut function inhibitory activity by using in vitro and in vivo assays. Ss.Cr caused a dose-dependent (0.03 - 3 mg/mL) inhibitory effect on K+-induced contractions in rat stomach fundus, guinea-pig ileum and rabbit jejunum preparations. The calcium channel blocking(CCB) activity was confirmed when Ss.Cr caused a rightward shift in the Ca++ dose-response curves. It also potentiated, at lower do-ses (0.001 - 0.03 mg/mL), the contractile effect of a fixed dose of acetylcholine (ACh), similar to physostigmine, and suppressed the effect of ACh at higher doses (0.3 - 1.0 mg/mL). Both Ss.Cr and physostigmine inhibited acetylcholinesterase (AChE), in the in vitro assay, confirming the AChE inhibitory activity. In the in vivo studies, Ss.Cr exhibited antidiarrheal and antisecretory activities against castor oil-induced diarrhea and intestinal fluid accumulation in mice. Characteristic steroidal compounds of the plant (saracocine, saracodine, saracorine and alkaloid-C), exhibited a similar combination of AChE inhibitory and CCB activities. Thus this study provides a sound mechanistic base for some of the traditional uses of the plant in hyperactive gut states, in addition to providing the first evidence for verapamil to possess additional AChE inhibitory activity. Furthermore, these characteristic compounds with dual activity may be good candidates for further studies on their usefulness in Alzheimer's disease. PMID- 15789499 TI - [Humeral shaft fractures in the adult]. PMID- 15789500 TI - [Isolated patello-femoral osteoarthritis]. PMID- 15789501 TI - [Development of professional responsibility in orthopedic surgery]. PMID- 15789502 TI - [Surgical treatment of rotator cuff rupture not treatable by suturing]. PMID- 15789503 TI - Getting inside depression and suicide ideation. One comprehensive screening approach targets patients 4 years and older. PMID- 15789504 TI - Abstracts of the 6th Nordic Congress on Disaster Medicine. Oslo, Norway, 2-5 June 2004. PMID- 15789505 TI - Dietary trans-fatty acids alter adipocyte plasma membrane fatty acid composition and insulin sensitivity in rats. AB - The present study was designed to investigate the effects of dietary trans-fatty acids (TFA) present in Indian vanaspati (partially hydrogenated vegetable oils) in comparison with saturated fatty acids (SFA) on adipocyte plasma membrane fatty acid composition, fluidity, and insulin action. The effects of 3% energy (% en) TFA was studied at 2% and 4% en of linoleic acid (18:2 n-6). WNIN male weanling rats were divided into 4 groups and fed casein-based diet containing 10% groundnut oil control (CON), palmolein (SFA), blend of vanaspati and safflower oil (3% en TFA and 2% en 18:2 n-6, TFA-1), or blend of vanaspati and safflower oil (3% en TFA and 4% en 18:2 n-6, TFA-2) for 12 weeks. Compared with CON, rats fed TFA and SFA diets had high levels of fasting plasma insulin and triglycerides. Both TFA- and SFA-fed groups had low levels of arachidonic acid (20:4 n-6) in adipocyte plasma membrane phospholipids. However, adipocyte plasma membrane fluidity decreased only in TFA-fed rats. Norepinephrine-stimulated lipolysis was high, whereas the antilipolytic effect of insulin and insulin stimulated glucose transport were low in the adipocytes of SFA- and TFA-fed rats. However, the extent of decrease in the antilipolytic effect of insulin and insulin-stimulated glucose transport was greater in TFA-fed rats. These findings suggest that diet providing approximately 10% en SFA (PUFA/SFA [P/S] ratio 0.2) decreased adipocyte insulin sensitivity in rats. In these diets, replacement of approximately 2% en SFA (16:0) and approximately 1% en monounsaturated fatty acid (18:1 cis) with TFA decreased adipocyte insulin sensitivity to a greater extent. However, increasing dietary 18:2 n-6 did not prevent or reduce the TFA-induced adipocyte insulin resistance. PMID- 15789506 TI - Current estimates from the National Health Interview Survey, 1994. PMID- 15789507 TI - Attention Deficit Disorder and Learning Disability: United States, 1997-98. AB - OBJECTIVES: This report presents national estimates of the prevalence of diagnosed Attention Deficit Disorder (ADD) and/or Learning Disability (LD) in U.S. children. Differences in the prevalence of these conditions are examined for children with selected sociodemographic characteristics. The occurrence of other health conditions and use of educational and health care services are contrasted for children with only ADD, those with only LD, those with both diagnoses, and those with neither diagnosis. METHODS: Estimates in this report are based on data from the National Health Interview Survey (NHIS), a national household survey of the civilian noninstitutionalized population of the United States. The analysis focuses on 8,647 children 6-11 years of age in the 1997 and 1998 NHIS. RESULTS: In 1997-98 over 2.6 million children 6-11 years of age were reported to have ever had a diagnosis of ADD or LD. A diagnosis of only ADD was reported for 3 percent of children, a diagnosis of only LD for 4 percent, and a diagnosis of both conditions for 4 percent. The prevalence of ADD with or without LD was greater for boys than for girls. Having health insurance was associated with a diagnosis of only ADD. Living in a low-income or mother-only family occurred more often among children with a diagnosis of LD. Children with LD were nearly five times more likely to be in special education than children with a diagnosis of only ADD. Children with ADD, in contrast to children without this diagnosis, more often had contact with a mental health professional, used prescription medication regularly, and had frequent health care visits. PMID- 15789508 TI - Summary health statistics for the U.S. population: National Health Interview Survey, 1999. AB - OBJECTIVES: This report presents health statistics from the 1999 National Health Interview Survey (NHIS) for the civilian noninstitutionalized population of the United States, classified by age, sex, race and Hispanic origin, poverty status, family income, education, place of residence, region of residence, and, where appropriate, health insurance coverage. The topics covered are health status and limitations of activity, injuries and poisonings, health care access and utilization, and health insurance coverage. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics (NCHS), Centers for Disease Control and Prevention (CDC), and is representative of the civilian noninstitutionalized U.S. population. Data are collected during face-to-face interviews with adults present at the time of interview. Information about children and absent adults is obtained from an adult proxy respondent. SELECTED HIGHLIGHTS: Nearly 40% of Americans reported having excellent health in 1999, while almost 9% reported having either fair or poor health. Fourteen percent of the U.S. population did not have any health insurance coverage in 1999. Eighteen percent of single race non-Hispanic black persons and 32% of Hispanics were uninsured in 1999, compared with 11% of single race non-Hispanic white persons. Further, 47% of poor Hispanics and 43% of near poor Hispanics under age 65 years were uninsured; percents of uninsurance among poor and near poor single race non Hispanic white and black persons under age 65 years were much lower. Eighty percent of single race non-Hispanic white persons under age 65 years had private health insurance coverage, compared with 57% of single race non-Hispanic blacks and 49% of Hispanics in this same age category. PMID- 15789509 TI - Summary health statistics for the U.S. population: National Health Interview Survey, 1998. AB - OBJECTIVES: This report presents health statistics from the 1998 National Health Interview Survey (NHIS) for the civilian, noninstitutionalized population of the United States, classified by age, sex, race and Hispanic origin, poverty status, family income, education, place of residence, region of residence, and, where appropriate, health insurance coverage. The topics covered are health status and limitations of activity, injuries and poisonings, health care access and utilization, and health insurance coverage. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized U.S. population. Data are collected during face-to-face interviews with adults present at the time of interview. Information about children and absent adults is obtained from an adult proxy respondent. HIGHLIGHTS: Nearly 40% of Americans reported having excellent health in 1998, while almost 9% reported having either fair or poor health. Fifteen percent of the U.S. population did not have any health insurance coverage in 1998. Nineteen percent of non-Hispanic black persons and 33% of Hispanics were uninsured in 1998, as opposed to 11% of non-Hispanic white persons. Further, 46% of poor Hispanics and 44% of near-poor Hispanics under age 65 years were uninsured; percents of uninsurance among poor and near poor non-Hispanic white and black persons under age 65 years were much lower. Lastly, 80% of non-Hispanic white persons under age 65 years had private health insurance coverage, as opposed to 55% of non-Hispanic black persons and 49% of Hispanics in this same age category. PMID- 15789510 TI - Summary health statistics for U.S. children: National Health Interview Survey, 1998. AB - OBJECTIVES: This report presents statistics from the 1998 National Health Interview Survey (NHIS) on selected health measures for children under 18 years of age, classified by sex, age, race/ethnicity, family structure, parent's education, family income, poverty status, health insurance coverage, place of residence, region, and current health status. The topics covered are asthma, allergies, learning disabilities, Attention Deficit Disorder, use of medication, respondent-assessed health status, school-loss days, usual place of medical care, time since last contact with a health care professional, selected health care risk factors, and time since last dental contact. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized population of the United States. Data are collected during face-to-face interviews with adults present at the time of interview. Information about children is collected for one randomly selected child per family in face-to face interviews with an adult proxy respondent familiar with the child's health. HIGHLIGHTS: In 1998 most U.S. children under 18 years of age enjoyed excellent or very good health (84%). However, 12% of children had no health insurance coverage, and 6% of children had no usual place of medical care. Twelve percent of children had ever been diagnosed with asthma. An estimated 8% of children 3-17 years of age had a learning disability, and an estimated 6% of children had Attention Deficit Disorder (ADD). Lastly, 11% of children in single mother families had two or more visits to an emergency room in the past year compared with 6% of children in two-parent or single-father families. PMID- 15789511 TI - Summary health statistics for U.S. children: National Health Interview Survey, 1999. AB - OBJECTIVES: This report presents statistics from the 1999 National Health Interview Survey (NHIS) on selected health measures for children under 18 years of age, classified by sex, age, race/ethnicity, family structure, parent's education, family income, poverty status, health insurance coverage, place of residence, region, and current health status. The topics covered are asthma, allergies, learning disability, attention deficit disorder, use of medication, respondent-assessed health status, school-loss days, usual place of medical care, time since last contact with a health care professional, selected health care risk factors, and time since last dental contact. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized population of the United States. Basic information is collected during face-to-face interviews with adults present at the time of interview. Information about children is collected about one randomly selected child per family in face-to-face interviews with an adult proxy respondent familiar with the child's health. SELECTED HIGHLIGHTS: In 1999 most U.S. children under 18 years of age enjoyed excellent or very good health (83%). However, 12% of children had no health insurance coverage, and 6% of children had no usual place of medical care. Eleven percent of children had ever been diagnosed with asthma. An estimated 7% of children 3-17 years of age had a learning disability, and an estimated 6% of children had Attention Deficit Disorder (ADD). Lastly, 9% of children in single mother families had two or more visits to an emergency room in the past year, compared with 4% of children in two parent families. PMID- 15789512 TI - Regional air pollution at a turning point. AB - The control of transboundary air pollution in Europe has been successful. Emissions of many key pollutants are decreasing and there are signs of improvements in damaged ecosystems. The strategies under development within the CAFE programme under the European Commission and the Convention on Long-range Transboundary Air Pollution (CLRTAP), aim to take regional air pollution control a large step further, in particular with respect to small particles. In this paper we highlight the new strategies but look primarily at socioeconomic trends and climate change feedbacks that may have a significant influence on the outcome of the strategies and which so far have not been considered. In particular, we point out the influence on air quality of increased summer temperatures in Europe and of increasing emissions including international shipping, outside of Europe. Taken together the further emissions reductions in Europe and the increasing background pollution, slowly cause a greying of the Northern Hemisphere troposphere rather than the traditional picture of dominant emissions in Europe and North America ('black') with much lower emission intensities elsewhere ('white'). A hemispheric approach to further combat air pollution will become necessary in Europe and elsewhere. PMID- 15789513 TI - Comparative health impact assessment of local and regional particulate air pollutants in Scandinavia. AB - The ongoing program Clean Air for Europe (CAFE) is an initiative from the EU Commission to establish a coordinated effort to reach better air quality in the EU. The focus is on particulate matter as it has been shown to have large impact on human health. CAFE requested that WHO make a review of the latest findings on air pollutants and health to facilitate assessments of the different air pollutants and their health effects. The WHO review project on health aspects of air pollution in Europe confirmed that exposure to particulate matter (PM), despite the lower levels we face today, still poses a significant risk to human health. Using the recommended uniform risk coefficients for health impact assessment of PM, regardless of sources, premature mortality related to long range transported anthropogenic particles has been estimated to be about 3500 deaths per year for the Swedish population, corresponding to a reduction in life expectancy of up to about seven months. The influence of local sources is more difficult to estimate due to large uncertainties when linking available risk coefficients to exposure data, but the estimates indicate about 1800 deaths brought forward each year with a life expectancy reduction of about 2-3 months. However, some sectors of the population are exposed to quite high locally induced concentrations and are likely to suffer excessive reductions in life expectancy. Since the literature increasingly supports assumptions that combustion related particles are associated with higher relative risks, further studies may shift the focus for abatement strategies. CAFE sets out to establish a general cost effective abatement strategy for atmospheric particles. Our results, based on studies of background exposure, show that long-range transported sulfate rich particles dominate the health effects of PM in Sweden. The same results would be found for the whole of Scandinavia and many countries influenced by transboundary air pollution. However, several health studies, including epidemiological studies with a finer spatial resolution, indicate that engine exhaust particles are more damaging to health than other particles. These contradictory findings must be understood and source specific risk estimates have to be established by expert bodies, otherwise it will not be possible to find the most cost effective abatement strategy for Europe. We are not happy with today's situation where every strategy to reduce PM concentrations is estimated to have the same impact per unit change in the mass concentration. Obviously there is a striking need to introduce more specific exposure variables and a higher geographical resolution in epidemiology as well as in health impact assessments. PMID- 15789514 TI - Nitrogen deposition and the biodiversity of boreal forests: implications for the nitrogen critical load. AB - The critical load concept is used to establish the deposition levels which ecosystems can tolerate without significant harmful effects. Here we summarize work within the Swedish research program Abatement Strategies for Transboundary Air Pollution (ASTA) assessing the critical load of N for boreal forests. Results from both field experiments in an area with low background N deposition in northern Sweden, and from a large-scale monitoring study, show that important vegetational changes start to take place when adding low N doses and that recovery of the vegetation after ceasing N input is a very slow process. The data presented indicate that changes in key ecosystem components occur even at a lower rate of N input than the present recommended empirical critical load for boreal forest understorey vegetation of 10-15 kg N ha(-1) yr(-1). Based on the data presented, we suggest that the critical load should be lowered to 6 kg N ha(-1) yr(-1). PMID- 15789515 TI - Modeling recovery of Swedish ecosystems from acidification. AB - Dynamic models complement existing time series of observations and static critical load calculations by simulating past and future development of chemistry in forest and lake ecosystems. They are used for dynamic assessment of the acidification and to produce target load functions, that describe what combinations of nitrogen and sulfur emission reductions are needed to achieve a chemical or biological criterion in a given target year. The Swedish approach has been to apply the dynamic acidification models MAGIC, to 133 lakes unaffected by agriculture and SAFE, to 645 productive forest sites. While the long-term goal is to protect 95% of the area, implementation of the Gothenburg protocol will protect approximately 75% of forest soils in the long term. After 2030, recovery will be very slow and involve only a limited geographical area. If there had been no emission reductions after 1980, 87% of the forest area would have unwanted soil status in the long term. In 1990, approximately 17% of all Swedish lakes unaffected by agriculture received an acidifying deposition above critical load. This fraction will decrease to 10% in 2010 after implementation of the Gothenburg protocol. The acidified lakes of Sweden will recover faster than the soils. According to the MAGIC model the median pre-industrial ANC of 107 microeq L(-1) in acid sensitive lakes decreased to about 60 microeq L(-1) at the peak of the acidification (1975-1990) and increases to 80 microeq L(-1) by 2010. Further increases were small, only 2 microeq L(-1) between 2010 and 2040. Protecting 95% of the lakes will require further emission reductions below the Gothenburg protocol levels. More than 7000 lakes are limed regularly in Sweden and it is unlikely that this practice can be discontinued in the near future without adverse effects on lake chemistry and biology. PMID- 15789516 TI - Economic assessment of the negative impacts of ozone on crop yields and forest production. A case study of the estate Ostads Sateri in southwestern Sweden. AB - Ground level ozone concentrations, in combination with the prevailing climate, at the estate Ostads Sateri in southwestern Sweden were estimated to reduce the yield of wheat and potato ranging between 5% and 10%. Occasionally, in years with the highest ozone concentrations and/or climatic conditions favoring high rates of ozone uptake to the leaves, yield loss levels above 10% may occur. Based on simple extrapolation, these ozone-induced reductions of crop yields at Ostads Sateri represent a potential total annual yield loss in Sweden in the range of 24.5 million Euro for wheat and 7.3 million Euro for potato, respectively. A simulation of forest growth at Ostad Sateri predicted that prevailing mean ozone exposure during 1993-2003 had the potential to reduce forest growth by 2.2% and the economic return of forest production by 2.6%. Using this value for extrapolation to the national level, the potential annual economic loss for Sweden due to negative impacts of ozone on forest production would be in the range of 56 million Euro (2004 prices). PMID- 15789517 TI - Nonlinearities in source receptor relationships for sulfur and nitrogen compounds. AB - The relationship between emissions and deposition of air pollutants, both spatially and in time forms an important focus for science and for policy makers. In practice, this relationship may become nonlinear if the underlying processes change with time, or in space. Nonlinearities may also appear due to errors in emission or deposition data, and careful scrutiny of both data sources and their relationship provides a means of picking up such deficiencies. Nonlinearities in source receptor relationships for sulfur and nitrogen compounds in Europe have been identified in measurement data for the UK. In the case of sulfur, the dry deposition process has been shown to be strongly influenced by ambient concentrations of NH3, leading to substantial increases in deposition rate as SO2 concentrations decline and the ratio SO2/NH3 decreases. The field evidence extends to measurements over three different surfaces in three countries across Europe. A mechanistic understanding of the cause of this nonlinearity has been provided. Apparent nonlinearities also exist in the sulfur deposition field through the influence of shipping emissions. The effect is clear at west coast locations, where during a period in which land-based sulfur emissions declined by 50%, no significant decline in concentrations of SO(2-) in precipitation were observed. The sites affected are primarily the coastal regions of southwestern UK, where shipping sources contribute a substantial fraction of the deposited sulfur, but the effect is not detectable elsewhere. Full quantification of the spatially disaggregated emission and their changes in time will eliminate this apparent nonlinearity in the source-receptor data. For oxidized nitrogen emission and deposition in the UK, there is strong evidence of nonlinearity in the source receptor relationship. The concentrations and deposition of NO(3-) in precipitation have declined little following a reduction in emissions of 45% during the period 1987 to 2001. The data imply a significant decrease in the average transport distance for oxidized nitrogen and most probably an increase in the average oxidation rate. However, the net effect of changes in aerosol chemistry due to changes in sulfur emissions and less competition for the main oxidants as a consequence of reductions in sulfur emission have not been separated. A quantitative explanation of the cause of this nonlinearity is lacking and the effects are therefore identified as an important uncertainty for the development of further protocols to control acidification, eutrophication and photochemical oxidants in Europe. PMID- 15789518 TI - European abatement of surface ozone in a global perspective. AB - EU's programme Clean Air for Europe (CAFE) is presently revising the policy on air quality which will lead to the adoption of a thematic strategy on air pollution under the Sixth Environmental Action Programme by mid-2005. For the abatement of surface ozone it is becoming evident that processes outside European control will be crucial for meeting long-term aims and air quality guidelines in Europe in the future. Measurements and modelling results indicate that there is a strong link between climate change and surface ozone. A warmer and dryer European climate is very likely to lead to increased ozone concentrations. Furthermore, increased anthropogenic emissions in developing economies in Asia are likely to raise the hemispheric background level of ozone. A significant increase in the background concentration of ozone has been observed at several sites in Northern Europe although the underlying causes are not settled. The photochemical formation of tropospheric ozone from increased concentrations of methane and CO may also lead to a higher ozone level on a global scale. Gradually, these effects may outweigh the effect of the reduced European ozone precursor emissions. This calls for a global or hemispheric perspective in the revision of the European air quality policy for ozone. PMID- 15789519 TI - The contribution from shipping emissions to air quality and acid deposition in Europe. AB - A global three-dimensional Lagrangian chemistry-transport model STOCHEM is used to describe the European regional acid deposition and ozone air quality impacts along the Atlantic Ocean seaboard of Europe, from the SO2, NOx, VOCs and CO emissions from international shipping under conditions appropriate to the year 2000. Model-derived total sulfur deposition from international shipping reaches over 200 mg S m(-2) yr(-1) over the southwestern approaches to the British Isles and Brittany. The contribution from international shipping to surface ozone concentrations during the summertime, peaks at about 6 ppb over Ireland, Brittany and Portugal. Shipping emissions act as an external influence on acid deposition and ozone air quality within Europe and may require control actions in the future if strict deposition and air quality targets are to be met. PMID- 15789520 TI - Expanded knowledge: the challenge to residency programs. PMID- 15789521 TI - Upper extremity peripheral nerve injury. PMID- 15789522 TI - Managing torus fracture of the radius. PMID- 15789523 TI - Percutaneous reduction and Kirschner-wire fixation of impacted intra-articular fractures and volar lip fractures of the proximal interphalangeal joint. AB - We retrospectively review the cases of 5 men with closed, impacted intra articular fractures of the middle phalanx at the proximal interphalangeal joint and describe a new technique-involving percutaneous reduction and Kirschner-wire fixation-that minimizes surgical trauma, allows fixation for early range of motion, and provides results as good as, if not superior to, those obtained with existing techniques. PMID- 15789524 TI - What's new in hip fractures? Current concepts. AB - Hip fractures have been among the most studied injury patterns in adults. The number of hip fractures is increasing exponentially, and their treatment costs place great economic strain on society. Recently developed hip fracture treatments, emphasizing cost containment, deformity prevention, and evidence based medicine, are attempts to optimize patient outcomes. In this article, we outline some of these developments with respect to femoral neck and intertrochanteric fractures. PMID- 15789525 TI - Preoperative versus postoperative radiation therapy for soft-tissue sarcomas. AB - In this article, we report results from a retrospective consecutive series of 117 patients with soft-tissue sarcomas treated with adjuvant radiation and surgical resection at 2 associated institutions. Fifty-nine patients received preoperative radiation; 58 patients received postoperative radiation. Mean clinical follow-ups were 6.1 years and 8.4 years, respectively. Nineteen (32.2%) of 59 patients in the preoperative group and 17 (29.3%) of 58 patients in the postoperative group had wound complications (P = .89). Three (5.1%) of 59 sarcomas in the preoperative group and 7 (12.1%) of 58 sarcomas in the postoperative group recurred locally (P = .19). Improved local disease control and other potential advantages of using preoperative radiation make this a preferred adjuvant treatment. PMID- 15789526 TI - Varus derotational osteotomy for spastic hip instability: the roles of femoral shortening and obturator neurectomy. AB - Varus derotational osteotomy (VDRO) was used to treat 69 unstable hips in 38 patients with spastic quadriplegia. After a mean follow-up of 5.3 years, the total rate of failure as judged by either need for revision or persistent radiographic instability or dislocation was 26%. Overall outcomes were better for hips that had been more congruent (preoperative migration index < 60%) than for hips that had experienced more migration (P = .027), and more-congruent hips were 9 times more likely to be stable radiographically at follow-up (P < .001). The failure rate for VDRO combined with femoral shortening or obturator neurectomy was lower than that for VDRO alone (P = .035). VDRO is an appropriate option for treating less advanced hip subluxation and may be more reliable when combined with femoral shortening and/or obturator neurectomy. VDRO alone, however, may be insufficient for treating more advanced instability. PMID- 15789527 TI - Necrotizing fasciitis in diabetes mellitus: a subtle presentation. PMID- 15789528 TI - A distal third clavicle fracture with posteroinferior displacement into the supraspinatus fossa. PMID- 15789529 TI - Anatomical risks of using supra-acetabular screws in percutaneous internal fixation of the acetabulum and pelvis. AB - We conducted a study to determine whether a lag screw placed percutaneously at the level of the pelvic brim for treatment of iliac fracture risks injury to the lateral femoral cutaneous nerve (LFCN). A 4-mm Kirschner wire (K-wire) was placed percutaneously into each of 8 human cadaveric hemipelvises (4 pelvises) at the level of the pelvic brim to represent the path of screw placement. Under fluoroscopic guidance, each K-wire was advanced from the anteroinferior iliac spine toward the posterior iliac crest. Cadavers were dissected at study end. Proximity of the LFCN to the percutaneously inserted K-wire was the main outcome measured. In 4 of the 8 hemipelvises, the LFCN was disrupted; in 3 hemipelvises, it was within 4 mm of the K-wire; in the last hemipelvis, it was 23 mm away. LFCNs varied anatomically from 1 to 5 branches; disruptions occurred more in LFCNs with multiple branches than in those with 1 branch. The results suggest considerable risk for injury to the LFCN during percutaneous fixation of iliac and acetabular fractures using a percutaneous screw at the level of the pelvic brim. PMID- 15789530 TI - Use of direct introduction of allograft anchors for rotator cuff repair. AB - Allograft anchors that can be inserted directly through the greater tuberosity were developed, tested, and used clinically for rotator cuff repair. Mechanical strength testing was performed, and allograft anchors were used in 63 consecutive procedures by a single surgeon. The insertion technique involved directly impacting the allograft anchor through the greater tuberosity without drilling a hole in the bone. Results of mechanical testing showed strengths comparable with those of other anchors (155.8 N), and the anchors locked consistently. This direct insertion technique saves time and procedural steps over traditional anchor insertion techniques while using an allograft anchor that reincorporates into host bone. PMID- 15789531 TI - [The necessary coordination of pancreas, islets and kidney transplantation in Spain]. PMID- 15789532 TI - [Fisiopathology of iron metabolism: diagnostic and therapeutic implications]. PMID- 15789533 TI - [Post-transplant lympho-proliferative disease associated to Epstein-Barr virus]. PMID- 15789534 TI - [Proteinuria and chronic renal failure in the coast of El Salvador: detection with low cost methods and associated factors]. AB - OBJECTIVES: El Salvador has high mortality rates from chronic kidney disease (CKD). The actual prevalence and causes remain unknown and local resources are scarce. Previous studies have suggested very high prevalence in specific populations: adult male farmers living in the coastlands, with high frequency of pesticides exposure and alcohol consumption. This population has low incomes and poor healthcare accessibility. Our aim was to detect CKD cases in this population using proteinuria as an inexpensive initial screening, investigating associations with those characteristics and comparing them with another population from the midlands. METHODS: We performed a transversal study on volunteer men from both regions, taking into consideration the variables: age, farmer living, pesticides, exposure, alcohol consumption, medical past history, blood pressure level, glycemia and proteinuria in a random urine sample. Coastland men with proteinuria were additionally screened for CKD measuring hemoglobin and serum creatinine levels. Finally, we employed a logistic regression model, and Perason's Chi2 to investigate associations between those variables and the presence of proteinuria or CKD. RESULTS: We included 291 men from the coastlands and 62 from the midlands: 35 and 16% off the total male population from their respective communities. Proteinuria (table I) was found in 133 (45.7%) and 8 cases (12.9%). CKD was diagnosed in 37 (12.7%) coastland men (table III), with mean creatinine 2.64 +/- 2.5 mg/dl, hemoglobin 12.32 +/- 1.9 g/dl and 81.1% with proteinuria 15 30 mg/l. Only 14 (37.8%) out of those 37 CKD patients featured diabetes or hypertension, while the remaining (62.2%) did not appear to have a clear-cut cause for CKD. Only one of them was formerly diagnosed with CKD. Farmer living, pesticides exposure and alcohol consumption were found to be very common characteristics in both populations, and were not associated to the presence of proteinuria or CKD (table II and III) DISCUSSION: The prevalence of CKD within the adult male farmers from the Salvadoran coastlands is remarkably high: at least 12.7%. There is a large number of undiagnosed cases, but they can be easily detected with inexpensive methods. This high prevalence is not completely explained by usual CKD causes like diabetes or hypertension. In addition, pesticides exposure and alcohol consumption may not be related, too. The disease is moderate, non symptomatic and has fairly mild proteinuria, possibly from interstitial origin. Further research is required to investigate environmental, occupational and hereditary factors, and to determine the real extent of the problem. PMID- 15789535 TI - [Tubulointerstitial nephritis associated with treatment with selective Cox-2 inhibitors, celecoxib and rofecoxib]. AB - The nephrotoxic effect of nonselective nonsteroidal anti-inflamatory drugs (NSAIDS) has been widely described. The main benefit of the Cox-2 inhibitors in relation to the NSAIDS is the production of a very similar analgesic effect, but with fewer gastrointestinal side effects. However, their effects on renal function are little known as yet and their long-term safety is still pending definition. The use of selective Cox-2 inhibitors as anti-inflamatory analgesic is becoming more and more common in our environment. We report two cases of tubulointersticial nephritis confirmed by renal biopsy, associated with administration of the two Cox-2 inhibitors currently available on the market, celecoxib and rofecoxib. In both cases, we were talking about elderly women, with deterioration of the general condition and acute renal failure. In the former case, renal biopsy showed an acute tubulo-intersticial nephritis (TIN) so highly "variegated" in its histologic expression. In the second case, was associated with strong indications of chronicity. Treatment with steroid was initiated in both patients and improvement of renal function was observed. PMID- 15789536 TI - [Analysis of non-EU immigrant population within the hemodialys program of the region of Madrid]. AB - Many hemodialysis patients from foreign countries arrived to be dialized in Madrid. They cames from all around the world, especially South America and Morroco. This group is younger (46.2 + 14.2 years) than the control group. Most of them initiated dialysis without a previous Nephrological check-up (62 %). In the foreign group the primary renal diseases were similar to the control group, however they also had many other health unusual problems. Most of them (71 %) started hemodialysis without ever having permanent hemodialysis vascular access, and 25 % were not able to speak spanish, or other European languages. This caused many problems in administering correct treatment. PMID- 15789537 TI - [Benefits of every other day dialysis (EODD) without rest for 72 hours in patients with symptomatic cardiovascular disease]. AB - With the purpose to improve the clinical situation of nine hemodialysis patients who suffer from severe cardiovascular disease and are highly symptomatic after weekends without dialysis because of fluid overload, their dialysis schedule was changed from 5 hours in 3 sessions per week to 4 hours every other day sessions (EODD), avoiding 72 hours of interdialitic weekend period. In each patient, during 38 sessions previous to starting the EODD (stage 1: 3 months) and the 38 sessions in EODD, which followed the first month of this dialysis regime (stage 2), the frequency of the next incidences was registered (ratio in 348 sessions, in every stage, of this patients group): presence of dysnea and/or hypertension pre dialysis session, pre or intra dialysis angor, emergency sessions with hypotension and sessions without achieving predetermined dry-weight. During the EODD stage, sessions, with dysena, hypertension and pre or intra dialysis angor were reduced in 80% (p < 0.001); the incidence of sessions with hypotensive episode or sessions without achieving dry-weight decreased in a third. All patients experimented a considerable improvement in their clinical situation. In addition, the whole group reduced dry-weight and later regained it without presenting symptoms which had motivated EODD schedule. EODD schedule improves the clinical situation in patients with cardiopathy who would not do so when following previous schedule (which includes 48 hours without dialysis). PMID- 15789538 TI - [Monitoring protocol of native vascular accesses for haemodialysis]. AB - Vascular access failure is an important cause of morbidity and mortality for patients on haemodialysis. We have prospectively studied, with a 5 years follow up, a monitoring protocol of native vascular accesses, using the available methods in every haemodialysis unit. All the native vascular accesses, created from 1- 1998 to XII-2001, with a posterior follow up until XII-2002, were monitored. Monitoring was based on physical examination, dificulty for blood flow greater than 300 ml/min, and in a monthly basis: dynamic venous pressure, urea recirculation and urea kinetic model. Abnormalities suggestive of fistulogram were a priori defined. During the recruitment period, a total of 164 accesses were created in 144 patients. Of these only 3 were grafts, 28 native vascular accesses were never functioning (primary failure rate 17. 1%), and 127 native accesses created in 113 patients (age 63.3 +/- 12.4 years; 18 % diabetics), were monitored (83% cephalic vein). Monitoring findings indicated realization of fistulogram in 35% and percutaneus angioplasty in 25% of the accesses. In order to maintain patency, the surgical intervention rate was 0.03 procedures/patient/year, the radiological 0.10 and the total 0.13. During the 5 years of the study occurred 41 thrombosis episodes in 40 accesses (0.07 thrombosis/patient/year), with percutaneus repermeabilization in 30%. Primary (unassisted) survival was 30.3 months (Confidence Interval 95% 25.6, 35.0) and secondary (assisted) survival 42.8 months (Confidence Interval 95%: 38.7, 46.9). Logistic regression analysis showed that presence of a central catheter at the time of creating the vascular access posses a greater risk for thrombosis (Relative Risk 5.6 if in subclavian vein), whereas age, diabetes, time to canulation, number of previous accesses and anatomic type do not increase that risk. In conclusion, in an old haemodialysis population, with an important diabetes prevalence, it is possible to create functioning native vascular accesses in most of them. The monitoring protocol allowed the detection and posterior correction of stenosis in a great number of accesses. The main risk of thrombosis is the presence of a central catheter at the time of creating a vascular access. PMID- 15789540 TI - [Disseminated cryptococosis in renal transplant recipient]. AB - The incidence of cryptococosis ranges from 0.4-5.8% in renal transplant. Meningitis is the principal clinical manifestation, frequently with a subacute curse. In renal transplantation recipients, disseminated cryptococcosis appears as the more frequent presentation. We report a case of a 32 years old woman renal transplant recipient who presents altered mental status, headache and tremor during the month before her assessment to our hospital. Microbiological study was performed in cerebrospinal fluid and cryptococcus was isolated. She was treated with amphotericin B and 5 flucytosine. She developed refractory increased intracranial pressure and a lumboperitoneal derivation was necessary. Cryptococcosis must be considered as cause of meningitis in patients with renal transplant. The early diagnosis and treatment are fundamental due to high mortality of this pathology. PMID- 15789539 TI - [Parvovirus B9 infection in a renal transplant recipient. Diagnosis by detection of viral genome in peripheral blood]. AB - Parvovirus B19 can produce a picture known as pure red blood aplasia in recipients of solid organ. Occasionally the viruses cause decrease of the other blood cells, and various extra-hematologic manifestations. Common diagnosis is realised by bone marrow examination. The diagnostic value of the viral genome in the blood stream is not well defined. We reported the case of a male of 17 years of age, whose diagnosis was done by repeated determinations of the viral parvovirus B19 genome in peripheral blood. It was confirmed by a biopsy of the iliac crest. The patient was treated with unspecific IgG immunoglobulins, with complete recovery from the symptoms and signs. It did not have any recurrence of the disease. This case suggests that the realisation of PCR of Parvovirus B19 in renal transplant patients with pure red cell aplasia could be of greater interest in the diagnosis and monitoring of the disease. The detection of the viral genome could avoid the administration of unnecessary blood transfusions, and possibly the realization of bone marrow biopsy. PMID- 15789541 TI - [Seminal vesicle cysts and infertility in autosomal dominant polycystic kidney disease]. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a systemic hereditary disorder characterized by bilateral diffuse renal cysts. Extrarenal involvement is a well known manifestation of ADPKD. Cysts in the liver, pancreas, lung, spleen, oesophagus, ovary, testis, epididymis, prostate, thyroid, bladder, uterus, brain, paraespinal, and seminal vesicle have also been described. The occurrence of seminal vesicle cysts is often unrecognised. We report here a man with seminal vesicle cysts and azoospermia associated with ADPKD. Seminal vesicle cysts are not uncommon in ADPKD and in some cases it is associated with infertility. Ultrasound and computed tomography imaging were effective in documenting the underlying lesions non-invasively. Studies evaluating fertility in patients with seminal vesicle cysts and ADPKD are needed. PMID- 15789542 TI - [Nephrotic syndrome associated to celecoxib]. PMID- 15789543 TI - [Occult hepatitis B virus infection in patients treated with chronic hemodialysis]. PMID- 15789544 TI - [Femoral arteriovenous fistula and heart failure, controlled after removal of hemodialysis catheter]. PMID- 15789545 TI - [Nephrotic syndrome, renovascular arterial hypertension, and proximal tubulopathy]. PMID- 15789546 TI - Leaf essential oil composition of five Zanthoxylum species from Monteverde, Costa Rica. AB - The leaf essential oils from five species of Zanthoxylum (Rutaceae) from Monteverde, Costa Rica, have been obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry. The species examined include Z. fagara, Z. acuminatum, Z. melanostictum, Z. monophyllum, and an undescribed species. The most abundant classes of compounds found in Zanthoxylum leaf oils are acyclic and menthane monoterpenoids as well as simple alcohols, aldehydes, and ketones. In terms of molecular diversity, menthane and acyclic monoterpenoids, cadinane and mesocyclic sesquiterpenoids, and simple alcohols, aldehydes, and ketones dominate the essential oils of Zanthoxylum species. Monoterpenoids make up the majority of the mass of the leaf oils of Z. monophyllum, Z. acuminatum, Z. fagara, and Zanthoxylum sp. nov. Linalool, 4-terpineol, alpha-terpineol, and trans-2-hexenol, are found in all of the Zanthoxylum species examined in this study. PMID- 15789547 TI - A facile and improved synthesis of sildenafil (Viagra) analogs through solid support microwave irradiation possessing tyrosinase inhibitory potential, their conformational analysis and molecular dynamics simulation studies. AB - Herein, the synthesis of some analogs of sildenafil (Viagra) (21) is described, employing MW irradiations in key steps such as, SNAr reaction on important precursor bromopyrazole (7). Compound 7 was synthesized by the bromination followed by the amidation of readily available 1-methyl-3-propyl-1H-pyrazole-5 carboxylic acid (5). Compounds 9 and 10 were obtained as SNAr reaction products, apparently through the proposed dipolar high-energy transition states TS-1 and TS 2 under MW irradiation, respectively. In contrast, conventional heating failed to produce similar results, even after prolonged heating. Compound 10, upon chlorosulfonation followed by the coupling of various nucleophiles, yielded a series of compounds 12-20 as analogs of sildenafil (21). Compounds 12-21 were subjected to tyrosinase inhibition studies and SAR studies were carried out. This study reflected that the inhibition was enhanced with increase of carbon chain. In case of the compound 17, the -OH group was replaced with -CH2-CH2-OH with a resulting increase in inhibition against tyrosinase. Compound 17 was found to be more potent than the potent reference inhibitor LM and KA. The 2D and 3D hydrogen bonding descriptors that help to study QSPR were also calculated. Energetically most stable conformations of these compounds were analyzed. Their kinetic, potential and total energies were also calculated through MD simulation. PMID- 15789548 TI - Semisynthetic preparation of leucomycin derivatives: introduction of aromatic side chains by reductive amination. AB - A small library of leucomycin A7 derivatives was prepared by NaCNBH3/ZnCl2 mediated reductive amination of the C18 aldehyde moiety with a variety of lipophilic benzylamines and tested for antibiotic activity. PMID- 15789549 TI - HPLC-coupled spectroscopic techniques (UV, MS, NMR) for the structure elucidation of phthalides in Ligusticum chuanxiong. AB - Ligusticum chuanxiong Hort., a plant that is frequently used in traditional Chinese medicine, has been studied using HPLC-coupled spectroscopic techniques such as HPLC-UV, HPLC-MS as well as HPLC-NMR. With the aid of these modern spectroscopic techniques, the main constituents, namely senkyunolide A, butylphthalide, neocnidilide and Z-ligustilide, have been characterized and identified. Phthalide dimers, present in smaller amounts, have been identified by HPLC-UV and HPLC-MS analysis and compared with reference compounds. Stereochemical features of some phthalide monomers have been determined by detailed spectroscopic studies for the first time. PMID- 15789550 TI - Alkaloid and phenolic compounds of Galanthus caucasicus, Magnolia obovata, Cocculus laurifolius, and Veratrum lobelianum grown in Georgia. AB - A collection of plants from the country of Georgia (Galanthus caucasicus, Magnolia obovata, Cocculus laurifolius, and Veratrum lobelianum) has been studied for their alkaloid content. Our studies led to the isolation and identification of 24 alkaloids, three of which are being identified and reported for the first time. PMID- 15789551 TI - A structurally diverse heterocyclic library: a benzothiazepine-fused beta-lactam library derived from reaction of 2,3-dihydro-1,5-benzothiazepines and acyl chlorides: synthesis and stereochemistry. AB - A 1H-azeto[2,1-d][1,5]benzothiazepin-1-one, beta-lactam derivatives of dihydrobenzothiazepines library derived from reactions of 2,4-disubstituted 2,3 dihydro-1,5-benzothiazepines and acyl chlorides, including phthalimidoacetyl chloride, chloroacetyl chloride, dichloroacetyl chloride and phenoxyacetyl chloride, was built up through parallel solution-phase synthesis. Stereochemistry of 1H-azeto[2,1-d][1,5]benzothiazepin-1-ones in the reaction process is discussed. PMID- 15789552 TI - Solid-supported reagents and catalysts for the preparation of large ring compounds. AB - Parallel combinatorial synthesis in solution using immobilized reagents, catalysts, and scavengers has emerged as a powerful technique for the preparation of diverse libraries of compounds. This technique has only recently been applied to the synthesis of large-ring compounds. In this comprehensive review several strategies are presented and discussed, including Pd-catalyzed allylic alkylation, Stille-coupling, macrolactonization and macrolactamization using solid supported reagents and catalysts. In several cases site isolation has allowed operation, of these macrocyclization reactions in concentrated solution (pseudo-dilution effect). PMID- 15789553 TI - Phthalocyanines as an alternative to soluble polymer-supported platforms in organic synthesis. AB - Some small molecules, such as phthalocyanines, capable of exhibiting greatly enhanced capacities and acting as internal tags, are shown to be effective as replacements for polydisperse polyethylene glycols, in soluble polymer-supported type synthesis of some piperidine and piperazine derivatives. PMID- 15789554 TI - Solid-phase synthesis of model libraries of 3alpha,17beta-dihydroxy-16alpha (aminoethyl-N-substituted)-5alpha-androstanes for the development of steroidal therapeutic agents. AB - The solid-phase synthesis of 16alpha-derivatives of 5alpha-androstane-3alpha, 17beta-diol with one, two or three levels of molecular diversity was accomplished using the diethylsilyloxy linker. Libraries with one level of diversity (10 members) and two levels of diversity (40 members) were synthesized in a parallel fashion in good yields and acceptable HPLC purities for the majority of library members. Compounds with three levels of diversity (15 pools) were realized in a split and pool fashion to allow further deconvolution by the positional scanning method. The screening of the generated model libraries revealed interesting preliminary structure-activity relationships related to their antiproliferative activities on androgen-sensitive Shionogi cells. In the case of the two-level library, the presence of a hydrophobic amino acid at R1 (isoleucine (Ile) or phenylalanine (Phe)) and a six-membered ring (aromatic or not) at R2 seems an important requirement for activity. In the three-level library, the amino acid residues isoleucine and phenylalanine clearly provided a better antiproliferative activity than glycine (Gly) and proline (Pro). These model libraries will serve as basis for the generation of larger libraries of peptidosteroids toward the development of therapeutic agents. PMID- 15789555 TI - Strategies for the synthesis of novel indole alkaloid-based screening libraries for drug discovery. AB - A series of diverse indole-based chemotypes were synthesized from beta tetrahydrocarboline (beta-THC) scaffolds prepared from commercially and readily available tryptamines and alpha-ketoesters. Diversity can be generated within these chemotypes through the following strategies: (a) appendage of substituents to the beta-THC scaffold, prepared in situ or as a template, through further elaboration and (b) skeletal modifications to the beta-THC scaffold via ring forming or ring breaking reactions. The strategies described here are amenable to high throughput solution-phase parallel synthesis, providing access to novel indole-based screening libraries for drug discovery. PMID- 15789556 TI - Fluorous parallel synthesis of a hydantoin/thiohydantoin library. AB - Fluorous tagging strategy is applied to solution-phase parallel synthesis of a library containing hydantoin and thiohydantoin analogs. Two perfluoroalkyl (Rf) tagged alpha-amino esters each react with six aromatic aldehydes under reductive amination conditions. Twelve amino esters then each react with 10 isocyanates and isothiocyanates in parallel. The resulting 120 ureas and thioureas undergo spontaneous cyclization to form the corresponding hydantoins and thiohydantoins. The intermediate and final product purifications are performed with solid-phase extraction (SPE) over FluoroFlash cartridges, no chromatography is required. Using standard instruments and straightforward SPE technique, one chemist accomplished the 120-member library synthesis in less than five working days, including starting material synthesis and product analysis. PMID- 15789557 TI - Total asymmetric synthesis of (-)-Phenylhistine, (-)-Aurantiamine and related compounds. Part I. AB - A new general, short, and efficient strategy for the construction of dehydro diketopiperazines was developed. Horner-Emmons type coupling between a phosphinyl glycine ester and a formyl heterocycle is the key coupling reaction, which proceeds in good-to-excellent yields on several sterically-hindered substrates. Moreover, racemization of the parent L-amino acids is avoided as a result of the mild basic conditions used. The selection of the NH protective group of the formyl heterocycle was crucial. N-tosylated heterocycles proved ideal for this reaction sequence. Thus, the title compounds, (-)-Phenylhistine and (-) Aurantiamine, were prepared in high yield (four steps, 47% overall) and optical purity. Furthermore, the synthesis of unnatural derivatives including an indole analogue was successfully completed. PMID- 15789558 TI - Solid-phase total synthesis of (-)-Phenylhistine and (-)-Aurantiamine. Synthesis of a diverse dehydro-2,5-diketopiperazine library. Part II. AB - The preparation of solid supported glycine phosphonate and its utilization for the total synthesis of two natural products is presented. The proposed protocol combines diversity with accessibility and speed, which makes this scaffold suitable for automated parallel synthesis and combinatorial chemistry. The preparation of a small library of dehydro-2,5-diketopiperazines, combining several natural amino acids with diverse heterocycles (including thiazoles, pyridines, indoles and imidazoles), is also demonstrated. PMID- 15789559 TI - Discovery of structurally diverse natural product antagonists of chemokine receptor CXCR3. AB - The chemokines (CXCL9, CXCL10 and CXCL11) and associated CXCR3 receptor are expressed during the inflammatory process from multiple sclerosis, atherosclerosis or organ transplantation resulting in the recruitment of lymphocytes leading to tissue damage. It is hypothesized that blocking of the ligand/CXCR3 receptor interaction has potential to provide opportunity for development of agents that would block tissue rejection. In this paper, four classes of natural product inhibitors (IC50 ranging 0.1-41 microM) have been described that block the CXCR3 receptor interaction of IP-10 ligand. These include a cyclic thiopeptide (duramycin), polyketide glycosides (roselipins), steroidal glycosides (hypoglausin A and dioscin) and a novel alkyl pyridinium alkaloid that were isolated by bioassay-guided fractionation of the organic extracts derived from actinomycete, fungal, plant and marine sources and discovered using 125I IP-10/CXCR3 binding assay. Duramycin was the most potent with an IC50 of 0.1 microM. Roselipins 2A, 2B and 1A showed IC50 values of 14.6, 23.5, and 41 microM, respectively. Diosgenin glycosides dioscin, hypoglaucin A and kallstroemin D exhibited IC50 values of 2.1, 0.47 and 3 microM, respectively. A novel cyclic 3-alkyl pyridinium salt isolated from a sponge displayed a binding IC50 of 0.67 microM. PMID- 15789561 TI - Myxobacterial epothilones and tubulysins as promising anticancer agents. AB - Tubulin-binding agents play a pivotal role in current cancer therapy and there are many efforts in pre-clinical and clinical development of known and novel cytotoxic agents ongoing. In this article a known class, epothilones, as well as a novel class, tubulysins, are presented. PMID- 15789560 TI - Rational selection of structurally diverse natural product scaffolds with favorable ADME properties for drug discovery. AB - Natural product analogs are significant sources for therapeutic agents. To capitalize efficiently on the effective features of naturally occurring substances, a natural product-based library production platform has been devised at Aurigene for drug lead discovery. This approach combines the attractive biological and physicochemical properties of natural product scaffolds, provided by eons of natural selection, with the chemical diversity available from parallel synthetic methods. Virtual property analysis, using computational methods described here, guides the selection of a set of natural product scaffolds that are both structurally diverse and likely to have favorable pharmacokinetic properties. The experimental characterization of several in vitro ADME properties of twenty of these scaffolds, and of a small set of designed congeners based upon one scaffold, is also described. These data confirm that most of the scaffolds and the designed library members have properties favorable to their utilization for creating libraries of lead-like molecules. PMID- 15789562 TI - Molecular diversity of Artocarpus champeden (Moraceae): a species endemic to Indonesia. AB - A summary of results obtained in the study of natural products isolated from a Moraceae species Artocarpus champeden is presented. The various classes of 3 prenylflavonoids isolated and the biogenetical correlation between the metabolites are discussed. Some of the flavonoids exhibited strong cytotoxicity against murine leukemia P388 cell lines, suggesting that flavonoids derived from moraceous plants represent a plausible unexplored resource of novel antitumor leads. PMID- 15789563 TI - Macrocycles rapidly produced by multiple multicomponent reactions including bifunctional building blocks (MiBs). AB - Naturally occurring macrocycles often exhibit remarkable biological activities and, therefore, constitute an attractive starting point for diversity-oriented synthesis for lead discovery in drug development. Multicomponent reactions have been used for the introduction of chemical diversity in strategies towards macrocycle libraries, mostly by combinational synthesis of a linear precursor combined with a subsequent macrocyclization reaction. The Ugi reaction in particular may be used for the macrocyclization itself as well, and a library of natural product-like macrocycles can be constructed in a single step from simple precursors. The efficiency and versatility of both strategies is immense and is exemplarily illustrated by the construction of small libraries of cyclopeptide alkaloid derivatives and biaryl ether macrocycles. The syntheses of the latter compound group are examples of multiple multicomponent macrocyclizations including bifunctional building blocks (M3iB3 or MiB), of which the Ugi-MiBs and their variations are discussed in more detail. PMID- 15789564 TI - What can a chemist learn from nature's macrocycles?--a brief, conceptual view. AB - Macrocyclic natural products often display remarkable biological activities, and many of these compounds (or their derivatives) are used as drugs. The chemical diversity of these compounds is immense and may provide inspiration for innovative drug design. Therefore, a database of naturally occurring macrocycles was analyzed for ring size, molecular weight distribution, and the frequency of some common substructural motifs. The underlying principles of the chemical diversity are reviewed in terms of biosynthetic origin and nature's strategies for diversity and complexity generation in relation to the structural diversity and similarities found in the macrocycle database. Finally, it is suggested that synthetic chemists should use not only nature's molecules, but also nature's strategies as a source of inspiration. To illustrate this, the biosynthesis of macrocycles by non-ribosomal peptide synthetases and terpene and polyketide cyclases, as well as recent advances of these strategies in an integrated synthesis/biotechnology approach are briefly reviewed. PMID- 15789565 TI - Recent advances in 1,3-dipolar cycloaddition reactions on solid supports. AB - Solid-phase methods are of a great significance in organic synthesis. Recent developments of these methods are providing new ways to construct libraries of small organic molecules. Five-membered heterocyclic compounds, which can be utilized in a variety of applications, are formed in the 1,3-dipolar cycloaddition reaction between dipolarophiles and dipoles. This review deals with the solid-phase synthesis of heterocycles via [3+2] cycloaddition reaction. Cycloaddition reactions of polymer-bound dipoles and polymer-bound dipolarophiles and intramolecular solid-phase cycloadditions are discussed in separate sections. Reactions of dipolarophiles such as alkenes, alkynes, and imines with dipoles such as azomethine ylides, azomethine imines, nitrile imines, azides, nitrones, and nitrile oxides are described. The recent literature up to December 2003 is covered. PMID- 15789566 TI - A simple approach to the synthesis of highly functionalized pyrrole derivatives. AB - The reaction of dibenzoylacetylene and enaminocarbonyl compounds leads to 3 alkylidene-2,3-dihydro-1H-pyrrol-2-ol derivatives in nearly quantitative yields. The reaction of this heterocyclic system with alcohols in the presence of a catalytic amount of HCl produces highly functionalized pyrroles in good yields. PMID- 15789567 TI - Semiempirical QSAR study and ligand receptor interaction of estrogens. AB - Softness values E(n)+/+ of estrogen derivatives and softness values E(m)+/+ of receptor lysine, histidine, tyrosine and cysteine have been evaluated by Klopman equation. The required parameters for the solution of Klopman equation have been calculated with the help of PM3 method. The difference deltaE(nm)+/+ between E(n)+/+ and E(m)+/+ has been derived for QSAR study. The estrogen derivatives have been divided into four different sets on the basis of their structural similarities, and their biological activity taken from literature in terms of relative binding affinity (RBA). The QSAR study shows that, deltaE(nm)+/+ values provide good relationship with biological activity. PMID- 15789568 TI - A novel three-component reaction designed by the combinatorial method: heteroarenes, isothiocyanates and isocyanides. AB - The novel three-component reaction of isoquinoline with isothiocyanates and isocyanides leads to a variety of new imidazoisoquinolines. The zwitterionic ground state of these new ring systems is established by means of NMR and X-ray analysis. Use of phthalazine instead of isoquinoline gives access to imidazole annulated phthalazines. PMID- 15789569 TI - Microwave assisted IMDAF# reaction: microwave irradiation applied with success to cycloaddition reaction of N-propargyl-N-p-tolyl-N-2-furfurylamines. AB - The new tertiary furfurylamine with triple bond as a dienophylic part i.e. N-(5 methyl-2-furfuryl)-N-prop-2-ynyl-p-toluidine (1) was prepared and the intramolecular Diels-Alder reaction of the amine (1) was performed under microwave irradiation conditions and by heating a benzene solution of the amine under nitrogen. Comparing the results of the usual thermal and the MAOS reaction, we confirmed our expectations that MAOS could promote the outcome of IMDA reaction of the suitably N-substituted tertiary 2-furfuryl-amines. In the present example, N-p-tolyl-5-methyl-5,7a-dihydro-5,7a-epoxyisoindoline was obtained in much better yield and of higher purity. PMID- 15789570 TI - Peptide mimotopes of phomopsins: identification, characterization and application in an immunoassay. AB - Peptide mimotopes of plant-associated toxins offer the potential for improving analytical and diagnostic methodologies as well as providing candidates for potential protective vaccines against plant poisoning diseases. Monoclonal antibody (mAb) C3C11, which recognizes the antimicrotubule phomopsin mycotoxins, was used to isolate peptide mimics of phomopsin A from a random 15-mer phage display peptide library. A total of 46 clones were isolated that showed specific reactivity with the mAb. Amino acid sequence analysis revealed four different types of mimotope sequences, all of which contained a common motif V-A-L/V-C. Of the 46 clones isolated, 44 contained the motif V-A-L-C while 2 contained the V-A V-C motif. All four types of phage clones inhibited the reactivity of the mAb with phomopsin A in a competition ELISA. The clone with the mimotope sequence CTVALCNMYFGAKLD demonstrated the strongest binding. It was further shown that synthetic peptides containing these mimotope amino acid sequences were able to inhibit the mAb-phomopsin A interaction, indicating that the peptide mimotopes were responsible for the specific binding, independent of the phage framework. The results also suggest that the mimotope peptides bind to mAb C3C11 at the same site as phomopsin A. The application of recombinant phage particles carrying phomopsin mimotopes in immunoassay was evaluated and the results demonstrated approximately 100-fold increase in sensitivity in comparison with a conventional immunoassay using a chemically linked phomopsin-horseradish peroxidase conjugate. PMID- 15789571 TI - Synthesis of dialkyl 2,5-dihydro-5-oxo-1,2-azaphosphole-3,4-dicarboxylates. AB - The adducts produced in the reaction between triphenylphosphine and dialkyl acetylenedicarboxylates were trapped by arylsulfonyl isocyanates to produce dialkyl 2,5-dihydro-5-oxo-1,2-azaphosphole-3,4-dicarboxylates in good yields. PMID- 15789572 TI - Medical physicists should actively discourage institutions from advertising technologies such as IMRT and HDR brachytherapy in order to recruit patients. For the proposition. PMID- 15789573 TI - Medical physicists should actively discourage institutions from advertising technologies such as IMRT and HDR brachytherapy in order to recruit patients. Against the proposition. PMID- 15789574 TI - Blood perfusion and thermal conduction effects in Gaussian beam, minimum time single-pulse thermal therapies. AB - A previous analytical study has shown that the minimum obtainable treatment time for a single pulse that delivers a given thermal dose to a specified point at a specified time occurs when the temperature at that point is rapidly raised to its maximum allowable value. The present study extends that result by investigating the spatial distribution of thermal effects of a single Gaussian shaped focal zone pulse that reaches that maximum allowable temperature at the center point of the focal zone. Analytical solutions are obtained that separately include the effects of perfusion and conduction. This situation is analyzed for a conservative treatment strategy in which the desired thermal dose is delivered when the tumor cools down to basal conditions. The results show that for a specified thermal dose delivered by a spherical Gaussian beam with focal widths below approximately 4 mm, the maximum allowable temperature, the minimum obtainable treatment time, and the size of the treatment zone (as a percentage of the size of the Gaussian beam) are all independent of the tissue blood perfusion, and are only functions of the focal zone size. Conversely, for focal widths above approximately 20 cm, these results are independent of the focal width and are only functions of blood perfusion. Between these two sizes (where most practical treatments will occur, since single pulses with widths of <4 mm and >20 cm will be uncommon in practice) a transition zone exists in which both perfusion and conduction effects are important. Thus while it is possible to implement a truly perfusion-independent, single pulse thermal treatment by using focal widths of <4 mm, in practice many such pulses will be needed to treat most tumors. This is especially true since the nonlinear temperature/thermal dose relationship causes the width of the delivered dose distribution to be only approximately 25%-30% of the width of the focal zone. However, shorter overall treatment times can be obtained when multiple pulses are linked together by using larger focal zone sizes, but this gain in treatment time is accompanied by increased effects of perfusion, illustrating the conflict between attaining both perfusion independence and minimal treatment time for multiple-pulse thermal treatments. PMID- 15789575 TI - An artificial neural network for predicting the incidence of radiation pneumonitis. AB - A method to predict radiation-induced pneumonitis (RP) using an artificial neural network (ANN) was investigated. A retrospective study was applied to the clinical data from 142 patients who have been treated with three-dimensional conformal radiotherapy for tumors in the thoracic region. These data were classified, based on their treatment outcome, into two patient clusters: with RP (Np=26) and without RP (Np= 116). An ANN was designed as a classifier. To perform the classification, a patient-treatment outcome with RP was assigned a value of 1, and a patient treatment outcome without RP was assigned a value of -1. The input of the ANN was limited to the patient lung dose-volume data only. A volume vector (VD) that describes patient lung subvolumes receiving more than a set of threshold doses was used as the network input variable. A zero value was used as the threshold to set the output value into -1 or 1. Three ANNs (ANN_1, ANN_2, and ANN_3), each with three layers, were trained to perform this classification function and to show the effect of training data on the ANN performance. Radial basis function was applied as the hidden layer neuron activation function and a sigmoid function was selected as the output layer neuron function. Backpropagation with a conjugate gradient algorithm was used to train the network. ANN_1 was trained and tested by using the leave-one-out method. ANN_2 was trained by randomly selecting 2/3 of the patient data, and tested by the remaining 1/3 of the data. ANN_3 was trained by a user selecting 2/3 of the patient data, and tested by the remaining 1/3 of the data. The predictive accuracy was verified as the area under a receiver operator characteristic (ROC) curve. The correct classification rates of 73% for RP cases, and 99% for non-RP cases were obtained from ANN_1. The corresponding correct classification rates of 44% for RP cases, and 89% for non-RP cases were obtained from ANN_2. From the ANN_3 test phase, the corresponding correct classification rates of 55% for RP cases, and 95% non-RP cases were achieved. The area under ROC curve was 0.85+/ 0.05, 0.68+/-0.10, and 0.81+/-0.09 for ANN_1, ANN _2, and ANN_3, respectively, within its asymmetric 95% confidence interval. The sensitivity was 95%, 57%, and 71%, and the specificity was 94%, 88%, and 90% for ANN_1, ANN_2, and ANN_3, respectively. Preliminary results suggest that the ANN approach provides a useful tool for the prediction of radiation-induced lung pneumonitis, using the patient lung dose-volume information. PMID- 15789576 TI - Neutron-activation revisited: the depletion and depletion-activation models. AB - The growth of a radioactive daughter in neutron activation is commonly described with the saturation model that ignores the consumption of parent nuclei during the radio-activation process. This approach is not valid when radioactive sources with high specific activities are produced or when the particle fluence rates used are very high. Assuming a constant neutron fluence rate throughout the activation target, a neutron-activation model that accounts for the depletion in parent nuclei is introduced. This depletion model is governed by relationships similar to those describing the parent-daughter-granddaughter decay series, and, in contrast to the saturation model, correctly predicts the practical limit of the daughter specific activity, irrespective of the particle fluence rate. Also introduced is a neutron-activation model that in addition to parent depletion accounts for the neutron activation of daughter nuclei in situations where the cross section for this effect is high. The model is referred to as the depletion activation model and it provides the most realistic description for the daughter specific activity in neutron activation. Three specific neutron activation examples of interest to medical physics are presented: activation of molybdenum 98 into molybdenum-99 described by the saturation model; activation of cobalt-59 into cobalt-60 described by the depletion model; and activation of iridium-191 into iridium-192 described by the depletion-activation model. PMID- 15789577 TI - rf enhancement and shielding in MRI caused by conductive implants: dependence on electrical parameters for a tube model. AB - Radio frequency (rf) eddy-currents induced in implants made of conductive material might cause significant image artifacts in magnetic resonance imaging (MRI) such as shielding of the lumen of vascular stents. rf alteration near metal parts was assessed theoretically in the approximation of alternating current electrodynamics: The implant was modeled as tube with diameter d(o), resistance R, and reactance Y, constituting the secondary winding of a transformer. The transmitter coil of the scanner acted as primary winding and generated the linearly polarized rf field B1,app. Tube axis was assumed parallel to B1,app. The results of the calculations were as follows: Ninety percent of the applied rf field amplitude is reached in the lumen at a ratio chi=R/Y approximately 2. A rapid drop occurs with the reduction of chi, whereas a further increase of chi causes only a small effect. With chi approximately 1/d(o)(Y approximately d2o,R approximately d(o)), conditions for rf alteration clearly depend on the diameter of the tube. Inside tubes with smaller diameter, rf shielding is less pronounced. rf alteration increases in good approximation with the square root of the strength of the static field B0. The following experiments were carried out: Tubes of similar diameter (d(o) approximately 8 mm) made of material of different conductivity (Cu, Nitinol, carbon fiber reinforced plastic with three different fiber structures) were examined at B0=0.2 and 1.5 T in water phantoms. Tube axis was aligned perpendicular to B0 and spin-echo technique was applied. Local rf enhancement near the outer surface of the metal tubes was detected applying manual reduction of the transmitter amplitude. Shielding inside a carbon fiber tube with d(o) approximately 8 mm and inside a smaller tube with d(o)=3.3 mm was compared. Both tubes showed the same wall structure and thickness (d(w)=0.4 mm). All measurements confirmed the theoretical results. Consequences for the construction of vascular stents are discussed, as well as problems with image artifacts due to rf enhancement near solid conductive implants. PMID- 15789578 TI - Performance comparison of an active matrix flat panel imager, computed radiography system, and a screen-film system at four standard radiation qualities. AB - Four standard radiation qualities (from RQA 3 to RQA 9) were used to compare the imaging performance of a computed radiography (CR) system (general purpose and high resolution phosphor plates of a Kodak CR 9000 system), a selenium-based direct flat panel detector (Kodak Direct View DR 9000), and a conventional screen film system (Kodak T-MAT L/RA film with a 3M Trimax Regular screen of speed 400) in conventional radiography. Reference exposure levels were chosen according to the manufacturer's recommendations to be representative of clinical practice (exposure index of 1700 for digital systems and a film optical density of 1.4). With the exception of the RQA 3 beam quality, the exposure levels needed to produce a mean digital signal of 1700 were higher than those needed to obtain a mean film optical density of 1.4. In spite of intense developments in the field of digital detectors, screen-film systems are still very efficient detectors for most of the beam qualities used in radiology. An important outcome of this study is the behavior of the detective quantum efficiency of the digital radiography (DR) system as a function of beam energy. The practice of users to increase beam energy when switching from a screen-film system to a CR system, in order to improve the compromise between patient dose and image quality, might not be appropriate when switching from screen-film to selenium-based DR systems. PMID- 15789579 TI - Derivation of the distribution of extrafocal radiation for head scatter factor calculation. AB - Head scatter factors for high energy photon beams from linear accelerators can be modeled using a two-source model consisting of focal and extrafocal radiation. The focal radiation can be approximated as a point source, and the distribution of the extrafocal radiation is a two-dimensional (2D) radial symmetric function. Various methods, including analytical, Monte Carlo, and empirical trial functions, have been used to determine the radial symmetric function of extrafocal radiation distribution. This article describes a method for directly determining the extrafocal radiation distribution without assuming any empirical trial function. The extrafocal radiation distribution is determined with measured head scatter factors for rectangular fields defined by the lower jaw (X) fixed at 40 cm and the upper jaw (Y) varying from 3 to 40 cm. The derivatives of the measured head scatter factors, with respect to the Y jaw position projected in the plane of extrafocal radiation, are proportional to the one-dimensional (1D) projection (also called the line spread function) of the extrafocal radiation distribution. Two methods are used to solve the radial function of extrafocal radiation from the 1D projection. The first method uses a 2D filtered backprojection algorithm, originally developed for parallel beam computed tomography reconstruction, to directly derive the radial dependence of the extrafocal radiation distribution. The method has been applied to 6 and 18 MV photon beams from a Siemens linear accelerator and has been tested by comparing measured and calculated head scatter factors for square and rectangular fields. The second method uses a Fourier transform followed by a Fourier-Bessel transform to solve the problem. The distributions of extrafocal radiation derived from these two methods are virtually identical. PMID- 15789580 TI - Estimation of the axial wall strains induced by an arterial stenosis at peak flow. AB - In the last 30 years, thousands of basic or clinical studies have been devoted to atherosclerosis or to the problem of restenosis after angioplasty. In these studies, axial stresses in the vessel wall have received practically no attention, contrary to circumferential stress and purely biological aspects. Based on a recent article describing how arterial stenoses can induce a considerable increase in axial wall stress during flow systole in the region immediately proximal to the stenosis entrance, we have used a simple (theoretical) spring model and data available in the literature on the mechanical properties of arteries to investigate the relative wall elongations (axial strains) resulting from the systolic increases in axial stress generated by the stenosis. The model shows that high axial wall strains are tightly limited to the stenosis entrance if the axial wall forces generating the supplementary stress are strongly absorbed by the tissues surrounding the vessel. Inversely, if this absorption is weak, the zone of high strains extends over a longer vessel segment upstream of the stenosis entrance. The maximum strain value, which is always situated at the stenosis entrance, appears to be relatively independent of the presence or absence of surrounding tissues. The simulation also shows that in a 3 mm coronary artery presenting a 75% diameter stenosis, the axial strain at the stenosis entrance can exceed 10% at peak flow, depending on the respective axial elasticities of vessel wall and surrounding tissues. In a more severe stenosis, or in case of a pathologically high systolic pressure, the maximum strain value might even exceed 20%. Since abnormal axial strains have been shown to induce abnormal biological processes in smooth muscle cells cultures, it is quite conceivable that such axial strains are deleterious, at least in arterial segments whose length normally does not vary. PMID- 15789581 TI - Optimizing the automatic segmentation of the left ventricle in magnetic resonance images. AB - Automatic segmentation of the left ventricular (LV) myocardial borders in cardiovascular MR (CMR) images allows a significant speed-up of the procedure of quantifying LV function, and improves its reproducibility. The automated boundary delineation is usually based on a set of parameters that define the algorithms. Since the automatic segmentation algorithms are usually sensitive to the image quality and frequently depend heavily on the acquisition protocol, optimizing the parameters of the algorithm for such different protocols may be necessary to obtain optimal results. In other words, using a default set of parameters may be far from optimal for different scanners or protocols. For the MASS-software, for example, this means that a total of 14 parameters need to be optimized. This optimization is a difficult and labor-intensive process. To be able to more consistently and rapidly tune the parameters, an automated optimization system would be extremely desirable. In this paper we propose such an approach, which is based on genetic algorithms (GAs). The GA is an unsupervised iterative tool that generates new sets of parameters and converges toward an optimal set. We implemented and compared two different types of the genetic algorithms: a simple GA (SGA) and a steady state GA (2SGA). The difference between these two algorithms lies in the characteristics of the generated populations: "nonoverlapping populations" and "overlapping populations," respectively "nonoverlapping" population means that the two populations are disjoint, and "overlapping" means that the best parameters found in the previous generation are included in the present population. The performance of both algorithms was evaluated on twenty routinely obtained short-axis examinations (eleven examinations acquired with a steady-state free precession pulse sequence, and nine examinations with a gradient echo pulse sequence). The optimal parameters obtained with the GAs were used for the LV myocardial border delineation. Finally, the automatically outlined contours were compared to the gold standard- manually drawn contours by experts. The result of the comparison was expressed as a degree of similarity after a processing time of less than 72 h to a 59.5% of degree of similarity for SGA and a 66.7% of degree of similarity for 2SGA. In conclusion, genetic algorithms are very suitable to automatically tune the parameters of a border detection algorithm. Based on our data, the 2SGA was more suitable than the SGA method. This approach can be generalized to other optimization problems in medical image processing. PMID- 15789582 TI - A dual-energy technique for enhanced localization accuracy in intracavitary brachytherapy. AB - The orthogonal imaging method is commonly used for source localization in brachytherapy. In some cases, however, difficulty is encountered in determining the dummy sources because of the presence of either contrast materials or bony structures. We here offer a novel method for source localization utilizing a dual energy, radiographic technique. In this approach, two sets of orthogonal radiographic images (anterior-posterior and lateral views) are obtained using two different x-ray energies. Image processing (i.e., subtraction between two image sets) is carried out to enhance the source image. In a study performed using a laboratory developed pelvic phantom, it was demonstrated that the dual-energy method could significantly enhance the image quality of the dummy sources, and improve the achievable precision and relative accuracy in localization of source positions. When directly combined with digital imaging modalities, the dual energy method can be a useful technique to improve the accuracy in brachytherapy source localization from planar radiographs. PMID- 15789583 TI - A fourier analysis of the dose grid resolution required for accurate IMRT fluence map optimization. AB - We present a theoretical and empirical analysis of the errors associated with the spatial discretization of the dose grid employed in optimized intensity modulated radiation therapy (IMRT) treatment plans. An information theory based Fourier analysis of the accuracy of discrete representations of three-dimensional dose distributions is presented. When applied to beamlet-based IMRT dose distributions, the theory produces analytic integrals that can bound worst case aliasing errors that can occur regardless of the location and orientation of the dose grid. The predictions of this theory are compared to empirical results obtained by solving a linear-programming based fluence-map optimization model to global optimality. A reasonable agreement between worst case estimates and the empirical results is attributed to the fact that the optimization takes advantage of aliasing to produce an optimal plan. We predicted and empirically demonstrated that an isotropic dose grid with <2.5 mm spacing is sufficient to prevent dose errors larger than a percent. However, we noted that in practice this resolution is mostly needed in high-dose target regions. Finally, a multiresolution 2-4-6 mm spacing model was developed and empirically tested where these spacings were applied to targets, structures, and tissue, respectively. PMID- 15789584 TI - A preliminary dosimetric characterization of chemical vapor deposition diamond detector prototypes in photon and electron radiotherapy beams. AB - Three radiation detectors based on polycrystalline diamond films with different thickness and resistivity, obtained by microwave chemical vapor deposition, were tested to assess their suitability for relative dosimetry of photon and electron beams supplied by clinical linear accelerators. All samples showed a linear response as a function of the absorbed dose. The sensitivity per unit of detector sensitive volume spanned between 7 and 43 nC Gy(-1) mm(-3) with an applied electric field of 40 kV/cm. The dose rate dependence was evaluated following the Fowler theory and delta coefficient values between 0.95 and 1.00 were found for the three samples when polarized at 40 kV/cm. Percentage depth dose curves, output factors, and normalized dose profiles were determined for 6 and 10 MV photon beams and for 6 and 15 MeV electron beams. The results obtained with the diamond detectors were in good agreement with those obtained by reference detector measurements [all the data were within the experimental uncertainty of 1% (1sigma)]. PMID- 15789585 TI - Is the diaphragm motion probability density function normally distributed? AB - During radiotherapy treatment planning, the margins given to the clinical target volume to form the planning target volume accounts for internal motion and set-up error. Most margin formulas assume that the underlying distributions are independent and normal. Clinical data suggests that the set-up error probability density function (pdf) can be considered to have an approximately normal distribution. However, there is evidence that internal motion does not have a normal distribution. Thus, in general, a convolution of the two pdfs should be performed to determine the total geometric error. The goals of this article were to (1) determine if the internal motion pdf due to respiration can be characterized using a normal distribution, and (2) if not, determine if the total geometric uncertainty for combining internal motion and set-up error can be characterized by a normal distribution. Sixty fluoroscopy diaphragm motion data sets were obtained using three breathing training types: free breathing, audio instruction, and visual feedback. Diaphragm motion was used as a surrogate for liver and lung cancer motion. The data were analyzed with normality tests in the following groups: (1) single motion measurements, (2) combined motion measurements for each patient, and (3) combined motion measurements for all patients. Following this analysis, the diaphragm motion pdfs were convolved with a set-up error pdf, and the standard deviation of the set-up error pdf at which the total geometric error pdf became normal was determined. At set-up error standard deviation values of at least 0.27 and 0.1 cm for free breathing, 0.57 and 0.42 cm for audio instruction, and 0.55 and 0 cm for visual feedback, for single motion measurements and combined motion measurements for each patient, respectively, total geometric error pdfs became approximately normal. When the motion measurements for all the patients were combined, diaphragm motion pdfs were approximately normal for all feedback types. Therefore, for treatment planning purposes in the absence of individual patient measurements, the diaphragm motion pdf can be considered an approximately normal distribution. However, care should be taken when determining a margin based on individual patients measurements as the total geometric error will, in general, not be normally distributed. PMID- 15789586 TI - Dosimetric study using different leaf-width MLCs for treatment planning of dynamic conformal arcs and intensity-modulated radiosurgery. AB - This paper systematically studied the dosimetric difference between a 3 mm micro multileaf collimator (MLC), a 5 mm MLC, and a 10 mm MLC for stereotactic radiosurgery using the Brainscan treatment planning system. Thirty-four cases treated with the dynamic conformal arcs technique and 20 cases treated with the intensity modulated radiosurgery/fractionated radiotherapy (IMRS/ IMRT) technique were retrospectively studied. The conformity index, the percentage target coverage, and the dose-volume histogram (DVH) for organs-at-risk (OARs) were used for dosimetric analysis and comparison for different treatment techniques, target volumes, and treatment sites. For the dynamic conformal arcs technique, there were statistically significant differences in the conformity indices between different leaf-width MLCs. The ratio of the conformity indices between different MLCs depended on the target volume. The average conformity index ratios between the 5 mm MLC and the 3 mm MLC were 1.37+/-0.09, 1.12+/-0.04, 1.08+/-0.02 and 1.04+/-0.01, respectively, for patients with the target volume (V) in groups: (1) V< 1 cm3, (2) 1 cm3 < V< 8 cm3, (3) 8 cm3 < V< 27 cm3, and (4) V> 27 CC. The average conformity index ratios between the 10 and 3 mm MLCs were 2.00+/-0.33, 1.45+/-0.09, 1.28+/-0.09, and 1.18+/-0.05 for patients in these four volume groups, respectively. No statistically significant difference was found for the target coverage among different MLCs. For the IMRS/IMRT technique, the average conformity index and target coverage ratios were 1.01+/-0.05 and 1.00+/-0.02, respectively, between the 5 and 3 mm MLCs, and were 1.04+/-0.07 and 0.97+/-0.02, respectively, between the 10 and 3 mm MLCs. The 3 mm MLC showed slightly better overall OAR DVHs than the 5 and 10 mm MLCs, especially for the cranial site with small-volume OARs defined. The results suggest that for the dynamic conformal arcs technique, the narrower leaf-width MLC provides better dose conformity than the wider leaf-width MLCs. This advantage decreases when the target volume increases. For the IMRS/IMRT technique, the narrower leaf-width MLC could have better sparing of small OARs than the wider leaf-width MLC. PMID- 15789587 TI - On the comparison of FROC curves in mammography CAD systems. AB - We present a novel method for assessing the performance of computer-aided detection systems on unseen cases at a given sensitivity level. The sampling error introduced when training the system on a limited data set is captured as the uncertainty in determining the system threshold that would yield a certain predetermined sensitivity on unseen data sets. By estimating the distribution of system thresholds, we construct a confidence interval for the expected number of false positive markings per image at a given sensitivity. We present two alternative procedures for estimating the probability density functions needed for the construction of the confidence interval. The first is based on the common assumption of Poisson distributed number of false positive markings per image. This procedure also relies on the assumption of independence between false positives and sensitivity, an assumption that can be relaxed with the second procedure, which is nonparametric. The second procedure uses the bootstrap applied to the data generated in the leave-one-out construction of the FROC curve, and is a fast and robust way of obtaining the desired confidence interval. Standard FROC curve analysis does not account for the uncertainty in setting the system threshold, so this method should allow for a more fair comparison of different systems. The resulting confidence intervals are surprisingly wide. For our system a conventional FROC curve analysis yields 0.47 false positive markings per image at 90% sensitivity. The 90% confidence interval for the number of false positive markings per image is (0.28, 1.02) with the parametric procedure and (0.27, 1.04) with the nonparametric bootstrap. Due to its computational simplicity and its allowing more fair comparisons between systems, we propose this method as a complement to the traditionally presented FROC curves. PMID- 15789588 TI - Effects of region-of-interest length on estimates of myocardial ultrasonic attenuation and backscatter. AB - Measurements of tissue properties using an image-based technique that makes use of an external reference may have the potential for practical clinical implementation in echocardiography. The objective of this study was to quantify the ability of this technique to distinguish myocardial attenuation and backscatter properties for specific lengths of the region-of-interest (ROI). We chose to exploit the anisotropic properties of the myocardium as a model for distinguishing tissue with different acoustic properties. Excised lateral wall segments from seven healthy adult sheep hearts were imaged using a commercially available (Philips/ATL) clinical scanner operating in the fundamental imaging mode with a linear array (L 7-4). Statistical and receiver operating characteristic (ROC) analyses were used to evaluate the ability of the video signal analysis method to differentiate midmyocardial from subendocardial regions based on measurements of the acoustic properties for specific lengths of the ROI. Results demonstrate that the ability to distinguish tissue properties increases with ROI length for both slope of attenuation and backscatter coefficient measurements. Statistically significant differences were observed for measurements utilizing the ROI lengths as short as 0.4 cm with corresponding progressively increasing areas under the ROC curves for increasing ROI lengths. [NIH R37 HL40302] PMID- 15789589 TI - Photon counting computed tomography: concept and initial results. AB - A concept of a photon counting cone beam CT is proposed. The system uses a new Multi Slit Multi Slice (MSMS) cone beam acquisition geometry utilizing a linear array photon counting detectors. The MSMS cone beam acquisition is a direct analogy of the scanning multislit acquisition used in projection x-ray imaging. This geometry provides a CT imaging with dose efficient scatter rejection and allows for using available photon counting detectors. The microchannel plate (MCP) detector is proposed as a linear array photon counting detector for MSMS cone beam CT system. Initial testing of the MCP detector for CT application was performed. The field of view of the prototype MCP detector is 60 mm. A delay line position encoding electronics was used. The electronics has a single channel input for evaluation of events from the entire detector field of view. This limits the system count rate at 2 x 10(5) count/s. The spatial resolution of this detector is 80 microm FWHM at 40 kVp and 200 microm FWHM at 90 kVp tube voltages. The detector noise in CT projections is less than 1 count/pixel for the 80 microm pixel size. The CT projections contain quantum-limited and scatter free signal. Images of a contrast phantom and a small animal were acquired at 50 kVp and 80 kVp tube voltages. The CT numbers for different contrast elements were calculated for a given x-ray spectrum and compared with experimental values. The quantum efficiency of the current detector is 56% at 90 kVp, which is suboptimal because of the large channel diameter (25 microm) of these MCPs. The MCPs with smaller channels and higher efficiencies are being tested. The quantum efficiency was measured to be 70% for a new MCP with 5 microm channel diameter. Design parameters of a clinically applicable photon counting MSMS cone beam CT for breast imaging was evaluated. System uses 20 cm field of view MCP detectors based on 5 microm channel MCPs and high count rate ASIC electronics. It was concluded that the MSMS cone beam CT with a photon counting MCP detector is feasible for volume breast imaging. PMID- 15789590 TI - A comparative study of rectal dose histograms in prostate brachytherapy: some analytic and numerical results. AB - A cumulative dose histogram is the graph of an integral function integrated over a domain VCR3 and is dubbed the dose-volume histogram (DVH), the dose-surface histogram (DSH) or the dose-wall histogram (DWH), depending on the dimension and structure of the region V. This paper presents a comparative study of the three rectal dose histograms for sixty patients as well as for a cylindrical model of the rectum; in particular, the DSH and DVH for the cylindrical model with one point source are computed analytically in terms of elliptic integrals. The difference among the three relative dose histograms, averaged over the sixty patients, is less than 5%, whereas that between DVH and DWH for various wall thickness can be as large as 3-12 cm3 in the range 60-100 Gy. The paper also contains an error analysis using two simple models of the rectum, for which the true DSH and DWH can be computed via numerical integration, to evaluate the effect of digitization. The digitized computation agrees quite well with the pre digitization numerical integration, within 1% or 0.2 cm3, because of the low dose gradient effect near the rectum in prostate brachytherapy. PMID- 15789591 TI - Dosimetry validation of treatment room shielding design. AB - Intensity-modulated radiation therapy (IMRT) can lead to an increase in leakage radiation. The total number of monitor units (MUs) for IMRT is typically 2-5 times that for conventional treatments [the ratio of the two is used to derive the effective modulation scaling factor (MSFeff)]. Shielding calculations for IMRT can be done by applying the MSFeff to measured exposures under conservative conditions (standard beam setup 40 cm x 40 cm field, 45 degrees collimator angle) to account for the increased leakage. In this work, we verified this approach for two existing vaults housing a Siemens Primart 6 MV linac and a Varian 21Ex 10 MV linac. We measured the cumulative exposures at various locations around the vaults for typical IMRT cases and for the standard beam setup using the same MUs. For the standard beam setup, the IMRT gantry angles and eight equally spaced angles were used. Estimations of weekly exposures for IMRT were carried out using exposure rates measured under standard beam setup and the MSFeff averaged over 20 treatment cases. The accumulated exposures under realistic IMRT conditions were 30%-50% lower than the estimated values using equally spaced gantry angles except for two locations where the real IMRT leakage was higher than the estimated value by approximately 10%. Measurements using the same gantry angles yielded similar results. Our results indicate that it is adequate to use the MSFeff and previously measured exposures to estimate the leakage increase due to IMRT for an existing vault. Different approaches should be followed when considering primary or secondary barriers since the standard beam setup is overestimating the exposures behind primary barriers compared to IMRT. In such cases, a 10 cm x 10 cm field can be used for more accurate shielding evaluation. PMID- 15789592 TI - Optimization of radiobiological effects in intensity modulated proton therapy. AB - Today, inverse treatment planning for intensity modulated proton therapy (IMPT) usually employs a constant relative biological effectiveness (RBE). In this paper, the potential clinical relevance of RBE variations for scanning techniques in IMPT is investigated, and a new strategy to include the RBE into the inverse planning process is presented. Three-dimensional RBE distributions are calculated based on a phenomenological model that describes the RBE as a function of dose, linear energy transfer (LET) and tissue type in the framework of the linear quadratic model. This RBE model is integrated into the optimization loop of inverse planning by using a modified version of the standard quadratic objective function, where the physical dose is replaced by the biological effect. This system for "biological optimization" was implemented into a research version of the inverse planning software KonRad and allows the direct optimization of the product of RBE and physical dose. Several treatment plans for a prostate case are presented, which compare the biological with the conventional physical dose optimization for IMPT scanning techniques, in particular distal edge tracking (DET) and the full three-dimensional (3D) modulation of beam spots. Mainly due to their different LET distributions, the RBE effects for these two techniques are quite different: while the RBE distribution was more or less homogeneous in the planning target volume (PTV) for 3D modulation, considerable RBE variations within the PTV were observed for DET. These unfavorable effects could be compensated for by employing the new biological objective function, which led to a more homogeneous distribution of the product of RBE and physical dose in the PTV. The computation time increased by a factor of 2 compared to the optimization of the physical dose. In conclusion, the proposed method allows the simultaneous multifield optimization of the biological effect in a reasonable time, and is therefore well suited for studying the influence of a variable RBE in IMPT as well as for minimizing potentially adverse effects. PMID- 15789593 TI - An Active Matrix Flat Panel Dosimeter (AMFPD) for in-phantom dosimetric measurements. AB - An a-Si Active Matrix Flat Panel Imager (AMFPI) prototype developed in-house has been modified to function as an in-phantom dosimetry system providing high resolution two-dimensional (2-D) data. This Active Matrix Flat Panel Dosimeter (AMFPD) system can be used as a replacement device for standard in-phantom dosimeters, such as scanning ion chambers in water, or film in solid water. The initial characterization of the device demonstrates a wide dynamic range (up to 160 cGy), a stable calibration curve (less than 1.5% variation over 1 year), dose rate independence (less than 1%), and excellent agreement of output factors with ion chamber measurements for a range of field sizes (less than 2%). The device also compares well to film for 2-D planar dose distributions. It is expected that the AMFPD system will be useful for beam commissioning, algorithm verification test data, and routine IMRT quality assurance dosimetry. PMID- 15789594 TI - Use and uncertainties of mutual information for computed tomography/ magnetic resonance (CT/MR) registration post permanent implant of the prostate. AB - Post-implant dosimetric analysis for permanent implant of the prostate benefits from the use of a computed tomography (CT) dataset for optimal identification of the radioactive source (seed) positions and a magnetic resonance (MR) dataset for optimal description of the target and normal tissue volumes. The CT/MR registration process should be fast and sufficiently accurate to yield a reliable dosimetric analysis. Since critical normal tissues typically reside in dose gradient regions, small shifts in the dose distribution could impact the prediction of complication or complication severity. Standard procedures include the use of the seed distribution as fiducial markers (seed match), a time consuming process that relies on the proper identification of signals due to the same seed on both datasets. Mutual information (MI) is more efficient because it uses image data requiring minimal preparation effort. A comparison of MI registration and seed-match registration was performed for twelve patients. MI was applied to a volume limited to the prostate and surrounding structures, excluding most of the pelvic bone structures (margins around the prostate gland were approximately 2 cm right-left, approximately 1 cm anterior-posterior, and approximately 2 cm superior-inferior). Seeds were identified on a 2 mm slice CT dataset using an automatic seed identification procedure on reconstructed three dimensional data. Seed positions on the 3 mm slice thickness T2 MR data set were identified using a point-and-click method on each image. Seed images were identified on more than one MR slice, and the results used to determine average seed coordinates for MR images and matched seed pairs between CT and MR images. On average, 42% (19%-64%) of the seeds (19-54 seeds) were identified and matched to their CT counterparts. A least-squares method applied to the CT and MR seed coordinates was used to produce the optimum seed-match registration. MI registration and seed match registration angle differences averaged 0.5 degrees, which was not significantly different from zero. Translation differences averaged 0.6 (1.2 standard deviation) mm right-left, -0.5(1.5) mm posterior-anterior, and 1.2(2.0) mm inferior-superior. Registration error estimates were approximately 2 mm for both the MI and seed-match methods. The observed standard deviations in the offset values were consistent with propagation of error. Registration methods as applied here using mutual information and seed matching are consistent, except for a small systematic difference in the inferior-superior axis for a minority of cases (approximately 15%). Cases registered with mutual information and with bony anatomy misregistration of greater than approximately 5 mm should be evaluated for rescan or seed-match registration. The improvement in efficiency of use for the MI registration method is substantial, approximately 30 min compared to several hours using seed match registration. PMID- 15789595 TI - The effect of gantry angle on megavoltage photon beam attenuation by a carbon fiber couch insert. AB - The use of rigid carbon fiber couch inserts in radiotherapy treatment couches is a well-established method of reducing patient set-up errors associated with couch sag. Several published studies have described such inserts as radiotranslucent with negligible attenuation of the radiation field. Most of these studies were conducted with the radiation field normally incident on the couch and there appears to be no evidence in the literature of the effect of the gantry angle on the extent of beam attenuation by the carbon fiber insert alone during external beam radiotherapy. In this study we examined the magnitude of this effect over a range of posterior oblique gantry angles using a cylindrical solid water phantom containing an ionization chamber placed isocentrically. It was found that a 6 MV photon beam, field size 10 x 5 cm, was attenuated significantly as the gantry angle approached the plane of the couch, from 2% at normal incidence and reaching 9% attenuation at angle of incidence 70 degrees. This could have serious implications regarding dose to the treatment volume for treatments requiring posterior oblique angles of incidence with a possible correction factor necessary in monitor unit calculations. PMID- 15789596 TI - Ghosting caused by bulk charge trapping in direct conversion flat-panel detectors using amorphous selenium. AB - Direct flat-panel detectors using amorphous selenium (a-Se) x-ray photoconductors are gaining wide-spread clinical use. The goal of our investigation is to understand the physical mechanisms responsible for ghosting, i.e., x-ray induced change in sensitivity that results in image persistence, so that the knowledge can be used to consistently minimize ghosting artifacts in a-Se flat-panel detectors. In this paper we will discuss the effect on x-ray sensitivity of charge trapping in a-Se, which is the dominant source for ghosting in a-Se flat panel detectors. Our approach is to correlate ghosting in electroded a-Se detectors with the trapped charge concentration measured by the "time-of-flight" (TOF) method. All measurements were performed as a function of radiation exposure X of up to approximately 20 R at electric field strength's of E(Se)=5 and 10 V/microm. The results showed that the x-ray sensitivity decreased as a function of X and the amount of ghosting decreased with increasing E(Se). The shape of the TOF curves changed as a result of irradiation in a manner indicating trapped electrons in the bulk of a-Se. The density of trapped electrons n(t) increases as a function of X. A method was developed to determine the values of n(t) in the bulk of a-Se from the TOF measurements, and to predict the corresponding change in x-ray sensitivity. Our results showed that a recombination coefficient consistent with that predicted by Langevin produced good agreement between calculated and measured x-ray sensitivity changes. Thus it can be concluded that the trapping of electrons in the bulk of a-Se and their subsequent recombination with x-ray generated free holes is the dominant mechanism for ghosting in a-Se. PMID- 15789597 TI - Regularization of inverse planning for intensity-modulated radiotherapy. AB - The performance of a variational regularization technique to improve robustness of inverse treatment planning for intensity modulated radiotherapy is analyzed and tested. Inverse treatment planning is based on the numerical solutions to the Fredholm integral equation of the first kind which is ill-posed. Therefore, a fundamental problem with inverse treatment planning is that it may exhibit instabilities manifested in nonphysical oscillations in the beam intensity functions. To control the instabilities, we consider a variational regularization technique which can be applied for the methods which minimize a quadratic objective function. In this technique, the quadratic objective function is modified by adding of a stabilizing functional that allows for arbitrary order regularization. An optimal form of stabilizing functional is selected which allows for both regularization and good approximation of beam intensity functions. The regularized optimization algorithm is shown, by comparison for a typical case of a head-and-neck cancer treatment, to be significantly more accurate and robust than the standard approach, particularly for the smaller beamlet sizes. PMID- 15789598 TI - Comparison of similarity measures for the task of template matching of masses on serial mammograms. AB - We conducted a study to evaluate the effectiveness of twelve different similarity measures in matching the corresponding masses on temporal pairs of current and prior mammograms. To perform this comparison we implemented each of the twelve similarity measures in the final stage of our multistage registration technique for automated registration of breast lesions in serial mammograms. The multistage technique consists of three stages. In the first stage an initial fan-shape search region was estimated on the prior mammogram based on the geometrical position of the mass on the current mammogram. In the second stage, the location of the fan-shape region was refined by warping, based on an affine transformation and simplex optimization. A new refined search region was defined on the prior mammogram. In the third stage, a search for the best match between the lesion template from the current mammogram and a structure on the prior mammogram was carried out within the search region. Our data set consisted of 318 temporal pairs. We performed three experiments, using a different subset of the 318 temporal pairs for each experiment. In each experiment we further tested how the performance of the similarity measures varied as the size of the search region increased or decreased. We evaluated the twelve similarity measures based on four criteria. The first criterion was the mean Euclidean distance, which was the average distance of the true location of the mass to the location detected by the similarity measure. The second criterion was the percentage of temporal pairs that were aligned so that 50% or more of the lesion area overlapped. The third criterion was the percentage of pairs that were aligned so that 75% or more of the lesion area overlapped. The fourth and final criterion was the robustness of the similarity measure. Our results showed that three of the similarity measures, Pearson's correlation, the cosine coefficient, and Goodman and Kruskal's Gamma coefficient, provide significantly higher accuracy (p < 0.05) in the task of matching the corresponding masses on serial mammograms than the other nine similarity measures. PMID- 15789599 TI - A dynamic renal phantom for nuclear medicine studies. AB - Dynamic radionuclide renal study (renography) provides functional and structural information of the kidney and urinary tract noninvasively. Our purpose in this study is to describe the construction and test results of a dynamic renal phantom with different clinical features of radionuclide renography. The phantom consisted of five pieces of different shaped Plexiglas boxes: Two kidneys, one liver, two square shaped boxes (one heart and one bladder). The bladder was internally divided into two compartments in order to collect each kidney output separately. The dynamic circulation of the phantom was maintained under a hydrostatic pressure approximately equal to 13.3 kPa (average human blood pressure). The standard dose distribution among different organs and different renographic parameters were calculated from series of normal patients study (91 with 99mTc-DTPA, 68 with 99mTc-EC). All the studies were performed with same camera (Siemens Orbiter Digitrac 7500) equipped with LEAP (low energy all purpose) collimator using ADAC Pegasys II analytic package program under the same clinical procedure. Different regions of interest (ROIs) were drawn for concerning organs and counts per second (CPS) were collected for each ROI. The series of renogram curves were generated by phantom-studies with different flow rates for left kidney (LK) and right kidney (RK). The renal index (RI) for an individual study was calculated as the product of two indexes: "Relative Renal Function" (RRF) (water-volume of LK/RK) and "Relative Renal Time" (RRT) (Tmax of LK/RK). The most significant correlation was found in total CPS for LK and RK between the EC group and phantom studies (p < 0.001). The calculated RI values were used to simulate the patients' study with different clinical features. The dynamics were found reproducible. The phantom is suitable for using in calibration and quality control protocols of the renogram procedure used in Nuclear Medicine. PMID- 15789600 TI - Dosimetric accuracy of Kodak EDR2 film for IMRT verifications. AB - Patient-specific intensity-modulated radiotherapy (IMRT) verifications require an accurate two-dimensional dosimeter that is not labor-intensive. We assessed the precision and reproducibility of film calibrations over time, measured the elemental composition of the film, measured the intermittency effect, and measured the dosimetric accuracy and reproducibility of calibrated Kodak EDR2 film for single-beam verifications in a solid water phantom and for full-plan verifications in a Rexolite phantom. Repeated measurements of the film sensitometric curve in a single experiment yielded overall uncertainties in dose of 2.1% local and 0.8% relative to 300 cGy. 547 film calibrations over an 18 month period, exposed to a range of doses from 0 to a maximum of 240 MU or 360 MU and using 6 MV or 18 MV energies, had optical density (OD) standard deviations that were 7%-15% of their average values. This indicates that daily film calibrations are essential when EDR2 film is used to obtain absolute dose results. An elemental analysis of EDR2 film revealed that it contains 60% as much silver and 20% as much bromine as Kodak XV2 film. EDR2 film also has an unusual 1.69:1 silver:halide molar ratio, compared with the XV2 film's 1.02:1 ratio, which may affect its chemical reactions. To test EDR2's intermittency effect, the OD generated by a single 300 MU exposure was compared to the ODs generated by exposing the film 1 MU, 2 MU, and 4 MU at a time to a total of 300 MU. An ion chamber recorded the relative dose of all intermittency measurements to account for machine output variations. Using small MU bursts to expose the film resulted in delivery times of 4 to 14 minutes and lowered the film's OD by approximately 2% for both 6 and 18 MV beams. This effect may result in EDR2 film underestimating absolute doses for patient verifications that require long delivery times. After using a calibration to convert EDR2 film's OD to dose values, film measurements agreed within 2% relative difference and 2 mm criteria to ion chamber measurements for both sliding window and step-and-shoot fluence map verifications. Calibrated film results agreed with ion chamber measurements to within 5 % /2 mm criteria for transverse-plane full-plan verifications, but were consistently low. When properly calibrated, EDR2 film can be an adequate two dimensional dosimeter for IMRT verifications, although it may underestimate doses in regions with long exposure times. PMID- 15789601 TI - Mass density images from the diffraction enhanced imaging technique. AB - Conventional x-ray radiography measures the projected x-ray attenuation of an object. It requires attenuation differences to obtain contrast of embedded features. In general, the best absorption contrast is obtained at x-ray energies where the absorption is high, meaning a high absorbed dose. Diffraction-enhanced imaging (DEI) derives contrast from absorption, refraction, and extinction. The refraction angle image of DEI visualizes the spatial gradient of the projected electron density of the object. The projected electron density often correlates well with the projected mass density and projected absorption in soft-tissue imaging, yet the mass density is not an "energy"-dependent property of the object, as is the case of absorption. This simple difference can lead to imaging with less x-ray exposure or dose. In addition, the mass density image can be directly compared (i.e., a signal-to-noise comparison) with conventional radiography. We present the method of obtaining the mass density image, the results of experiments in which comparisons are made with radiography, and an application of the method to breast cancer imaging. PMID- 15789603 TI - A method for determining the gantry angle for megavoltage cone beam imaging. AB - Accurate knowledge of gantry angle is essential in megavoltage cone beam imaging (MVCBI) with an electronic portal imager. We present a method for determining the gantry angle by detecting multileaf collimator (MLC) leaf positions in projection images. During image acquisition the gantry moves continuously and the MLC operates in dynamic arc mode. Our algorithm detects the leaf positions in the images and compares them with a stationary reference leaf. Comparison of the algorithm against angles determined from the locations of fiducial markers shows the accuracy (0.26 degrees rms error) to be sufficient for MVCBI. PMID- 15789602 TI - Segmented phosphors: MEMS-based high quantum efficiency detectors for megavoltage x-ray imaging. AB - Current electronic portal imaging devices (EPIDs) based on active matrix flat panel imager (AMFPI) technology use a metal plate+phosphor screen combination for x-ray conversion. As a result, these devices face a severe trade-off between x ray quantum efficiency (QE) and spatial resolution, thus, significantly limiting their imaging performance. In this work, we present a novel detector design for indirect detection-based AMFPI EPIDs that aims to circumvent this trade-off. The detectors were developed using micro-electro-mechanical system (MEMS)-based fabrication techniques and consist of a grid of up to approximately 2 mm tall, optically isolated cells of a photoresist material, SU-8. The cells are dimensionally matched to the pixels of the AMFPI array, and packed with a scintillating phosphor. In this paper, various design considerations for such detectors are examined. An empirical evaluation of three small-area (approximately 7 x 7 cm2) prototype detectors is performed in order to study the effects of two design parameters--cell height and phosphor packing density, both of which are important determinants of the imaging performance. Measurements of the x-ray sensitivity, modulation transfer function (MTF) and noise power spectrum (NPS) were performed under radiotherapy conditions (6 MV), and the detective quantum efficiency (DQE) was determined for each prototype SU-8 detector. In addition, theoretical calculations using Monte Carlo simulations were performed to determine the QE of each detector, as well as the inherent spatial resolution due to the spread of absorbed energy. The results of the present studies were compared with corresponding measurements published in an earlier study using a Lanex Fast-B phosphor screen coupled to an indirect detection array of the same design. The SU-8 detectors exhibit up to 3 times higher QE, while achieving spatial resolution comparable or superior to Lanex Fast-B. However, the DQE performance of these early prototypes is significantly lower than expected due to high levels of optical Swank noise. Consequently, the SU-8 detectors presently exhibit DQE values comparable to Lanex Fast-B at zero spatial frequency and significantly lower than Fast-B at higher frequencies. Finally, strategies for reducing Swank noise are discussed and theoretical calculations, based on the cascaded systems model, are presented in order to estimate the performance improvement that can be achieved through such noise reduction. PMID- 15789604 TI - Dose calibration of nonconventional treatment systems applied to helical tomotherapy. AB - Current dosimetric protocols based on the absorbed dose (AAPM TG-51 and IAEA TRS 398 protocols) require calibration measurements under reference conditions. For some radiotherapy systems, this requirement cannot be met, and calibration has to be performed under nonreference experimental conditions. In order to solve this problem, both protocols can be extended by inclusion of the measured-to-reference conversion factor, k(mr). In order to determine this factor, basic dosimetric quantities, like stopping power ratios, mass attenuation coefficients and chamber correction factors have to be calculated. If measurements are not feasible, accurate Monte Carlo modeling is required. The extension of the protocols is illustrated using the case of the helical tomotherapy radiation unit, where the typical calibration measurement conditions are the 10 x 5 cm2 field size and the 85 cm surface source distance, limited by the system design. It was calculated that the k(mr) factor for this conditions is close to unity (0.997+/-0.001). In addition, the deviation of the measurement conditions from the reference conditions results in the change of the quality conversion factor (approximately 0.995-0.998, depending on the ionization chamber used). This change is the same regardless of the used calibration protocol. For smaller field sizes the corrections become more significant, resulting in the total correction factor compared to the reference conditions of up to 1.5% for the smallest considered field size of 2 x 2 cm2. PMID- 15789605 TI - Evaluation of and compensation for spatial noise of LCDs in medical applications. AB - Recent developments in liquid crystal display (LCD) technology suggest that this technology will replace the cathode ray tube (CRT) as the most popular softcopy display technology in the medical arena. However, LCDs are far from ideal for medical imaging. One of the principal problems they possess is spatial noise contamination, which requires accurate characterization and appropriate compensation before LCD images can be effectively utilized for reliable diagnosis. This paper presents some work we have conducted recently on characterization of spatial noise of high resolution LCDs. The primary purpose of this work is to explore the properties of spatial noise and propose a method to reduce it. A high quality CCD camera was used for physical evaluation. Spatial noise properties were analyzed and estimated from the camera images via signal modeling and processing. A noise compensation algorithm based on error diffusion was developed to process images before they were displayed. Results shown in this paper suggest that LCD spatial noise can be effectively reduced via appropriate processing. PMID- 15789606 TI - Physical characterization of a prototype selenium-based full field digital mammography detector. AB - The purpose of this study was to measure experimentally the physical performance of a prototype mammographic imager based on a direct detection, flat-panel array design employing an amorphous selenium converter with 70 microm pixels. The system was characterized for two different anode types, a molybdenum target with molybdenum filtration (Mo/Mo) and a tungsten target with rhodium filtration (W/Rh), at two different energies, 28 and 35 kVp, with approximately 2 mm added aluminum filtration. To measure the resolution, the presampled modulation transfer function (MTF) was measured using an edge method. The normalized noise power spectrum (NNPS) was measured by two-dimensional Fourier analysis of uniformly exposed mammograms. The detective quantum efficiencies (DQEs) were computed from the MTFs, the NNPSs, and theoretical ideal signal to noise ratios. The MTF was found to be close to its ideal limit and reached 0.2 at 11.8 mm(-1) and 0.1 at 14.1 mm(-1) for images acquired at an RQA-M2 technique (Mo/Mo anode, 28 kVp, 2 mm Al). Using a tungsten technique (MW2; W/Rh anode, 28 kVp, 2 mm Al), the MTF went to 0.2 at 11.2 mm(-1) and to 0.1 at 13.3 mm(-1). The DQE reached a maximum value of 54% at 1.35 mm(-1) for the RQA-M2 technique at 1.6 microC/kg and achieved a peak value of 64% at 1.75 mm(-1) for the tungsten technique (MW2) at 1.9 microC/kg. Nevertheless, the DQE showed strong exposure and frequency dependencies. The results indicated that the detector offered high MTFs and DQEs, but structured noise effects may require improved calibration before clinical implementation. PMID- 15789607 TI - Three-dimensional IMRT verification with a flat-panel EPID. AB - A three-dimensional (3D) intensity-modulated radiotherapy (IMRT) pretreatment verification procedure has been developed based on the measurement of two dimensional (2D) primary fluence profiles using an amorphous silicon flat-panel electronic portal imaging device (EPID). As described in our previous work, fluence profiles are extracted from EPID images by deconvolution with kernels that represent signal spread in the EPID due to radiation and optical scattering. The deconvolution kernels are derived using Monte Carlo simulations of dose deposition in the EPID and empirical fitting methods, for both 6 and 15 MV photon energies. In our new 3D verification technique, 2D fluence modulation profiles for each IMRT field in a treatment are used as input to a treatment planning system (TPS), which then generates 3D doses. Verification is accomplished by comparing this new EPID-based 3D dose distribution to the planned dose distribution calculated by the TPS. Thermoluminescent dosimeter (TLD) point dose measurements for an IMRT treatment of an anthropomorphic phantom were in good agreement with the EPID-based 3D doses; in contrast, the planned dose under predicts the TLD measurement in a high-gradient region by approximately 16%. Similarly, large discrepancies between EPID-based and TPS doses were also evident in dose profiles of small fields incident on a water phantom. These results suggest that our 3D EPID-based method is effective in quantifying relevant uncertainties in the dose calculations of our TPS for IMRT treatments. For three clinical head and neck cancer IMRT treatment plans, our TPS was found to underestimate the mean EPID-based doses in the critical structures of the spinal cord and the parotids by approximately 4 Gy (11%-14%). According to radiobiological modeling calculations that were performed, such underestimates can potentially lead to clinically significant underpredictions of normal tissue complication rates. PMID- 15789608 TI - Generalizing the MTF and DQE to include x-ray scatter and focal spot unsharpness: application to a new microangiographic system. AB - Detector characterization with modulation transfer function (MTF) and detective quantum efficiency (DQE) inadequately predicts image quality when the imaging system includes focal spot unsharpness and patient scatter. The concepts of MTF, noise power spectrum, noise equivalent quanta and DQE were referenced to the object plane and generalized to include the effect of geometric unsharpness due to the finite size of the focal spot and the effect of the spatial distribution and magnitude of x-ray scatter due to the patient. The generalized quantities provide performance characteristics that consider the complete imaging system, but reduce to a description of the detector properties without magnification or scatter. We have evaluated a new neurovascular angiography imaging system based on a region of interest (ROI) microangiographic detector using these generalized quantities. A uniform head-equivalent phantom was used as a filter and x-ray scatter source. This allowed the study of all properties of the detector under clinically relevant x-ray spectra and x-ray scatter conditions. Realistic focal spots (0.8 mm nominal), beam energies (60-100 kVp), and detector exposures (0.8 2.3 mR) were used, and the effects of different scatter fractions (0-0.62) resulting from changing the beam size (0-100 cm2) were investigated. The generalized MTF and DQE were found to have very little dependence on the tube voltage and the detector entrance exposure. Magnification, with the focal spot used, results in a large decrease of the generalized DQE at higher frequencies (about 100-fold at 10 cycles/mm), but a significantly smaller decrease at lower frequencies. Scatter on the other hand, causes a constant drop in the generalized DQE (factor of 3 for scatter fraction 0.3) for all frequencies. Our results show that there are tradeoffs in the choice of the different system parameters; therefore this methodology of studying the imaging system as a whole could provide guidance in system design. PMID- 15789609 TI - Comparison of helical and cine acquisitions for 4D-CT imaging with multislice CT. AB - We proposed a data sufficiency condition (DSC) for four-dimensional-CT (4D-CT) imaging on a multislice CT scanner, designed a pitch factor for a helical 4D-CT, and compared the acquisition time, slice sensitivity profile (SSP), effective dose, ability to cope with an irregular breathing cycle, and gating technique (retrospective or prospective) of the helical 4D-CT and the cine 4D-CT on the General Electric (GE) LightSpeed RT (4-slice), Plus (4-slice), Ultra (8-slice) and 16 (16-slice) multislice CT scanners. To satisfy the DSC, a helical or cine 4D-CT acquisition has to collect data at each location for the duration of a breathing cycle plus the duration of data acquisition for an image reconstruction. The conditions for the comparison were 20 cm coverage in the cranial-caudal direction, a 4 s breathing cycle, and half-scan reconstruction. We found that the helical 4D-CT has the advantage of a shorter scan time that is 10% shorter than that of the cine 4D-CT, and the disadvantages of 1.8 times broadening of SSP and requires an additional breathing cycle of scanning to ensure an adequate sampling at the start and end locations. The cine 4D-CT has the advantages of maintaining the same SSP as slice collimation (e.g., 8 x 2.5 mm slice collimation generates 2.5 mm SSP in the cine 4D-CT as opposed to 4.5 mm in the helical 4D-CT) and a lower dose by 4% on the 8- and 16-slice systems, and 8% on the 4-slice system. The advantage of faster scanning in the helical 4D-CT will diminish if a repeat scan at the location of a breathing irregularity becomes necessary. The cine 4D-CT performs better than the helical 4D-CT in the repeat scan because it can scan faster and is more dose efficient. PMID- 15789610 TI - Comment on "an inverse-geometry volumetric CT system with a large-area scanned source: a feasibility study" [Med. Phys. 31, 2623-2627 (2004)]. PMID- 15789611 TI - In vivo PAF-induced airway eosinophil accumulation reduces bronchial responsiveness to inhaled histamine. AB - Chronic eosinophilic bronchitis and bronchial hyperresponsiveness have been considered to be the fundamental features of bronchial asthma. However, the role of airway eosinophils in bronchial responsiveness in vivo has not been fully discussed. The aim of this study was to investigate the direct effect of airway eosinophil accumulation on bronchial responsiveness in vivo. Guinea pigs were transnasally treated with platelet activating factor (PAF) or vehicle twice a week for a total of 3 weeks. Anesthetized guinea pigs were surgically cannulated and artificially ventilated 48 h after the last administration of PAF or vehicle. Ten minutes after the installation of artificial ventilation, ascending doses of histamine were inhaled. In a subsequent study, selective inhibitors of diamine oxidase and histamine N-methyltransferase were intravenously administered before the histamine inhalation in the PAF-treated animals. Next study was conducted 20 min after treatment with indomethacin in this study line. Finally, ascending doses of methacholine were inhaled in our animal model. Proportion of eosinophils and the number of nuclear segmentation in bronchoalveolar lavage fluid significantly increased in guinea pigs treated with PAF compared with vehicle and this finding was confirmed histologically. Nevertheless, bronchial responsiveness to inhaled histamine, but not methacholine, was significantly decreased by the PAF treatment. This bronchoprotective effect induced by PAF remained following aminoguanidine and histamine N-methyltransferase administration, but abolished by treatment of indomethacin. These results suggest that in vivo airway eosinophils may reduce nonspecific bronchial responsiveness through production of inhibitory or bronchoprotective prostanoids, but not through histaminase production. PMID- 15789612 TI - Effect of prostaglandin E2 on PMA-induced macrophage differentiation. AB - Major trauma such as severe bums and extensive surgery could result in accelerated macrophage differentiation and hyperactivation causing an excessive release of proinflammatory cytokines and prostaglandin E2 (PGE2) with consequent severe impairment of immunologic reactivity. HL-60 cells stimulated with phorbol 12-myristate 13-acetate (PMA) have been used as a model to asses the PGE2 role in the macrophage differentiation observed after major trauma. Cell adhesion, matrix metalloproteinase-9 (MMP-9) and tumor necrosis factor-alpha (TNF-alpha) production were measured after 24 h of PMA treatment in the presence of PGE2 (1 nM - 1 microM). PGE2 increased both the PMA-induced cell adhesion and MMP-9 production via EP2/EP4 receptors while it had no effect on the induced TNF-alpha release. The cAMP/PKA pathway, usually linked to EP2/EP4 activation, was not involved in the phenomenon, suggesting that an alternative signalling pathway could be linked to a PKC-activated enzyme. In fact PGE2 activity was partially inhibited by Wortmannin, a phosphoinositide-3 kinase (PI-3K) inhibitor indicating that PGE2 act as a co-factor able to increase macrophage differentiation in vitro via a PI-3K dependent pathway that could be also involved in the immunosuppression observed in the aftermath of trauma. PMID- 15789613 TI - Involvement of leukotriene B4 receptor 1 signaling in platelet-activating factor mediated neutrophil degranulation and chemotaxis. AB - Platelet-activating factor (PAF) is a potent lipid mediator of inflammation that can act on human neutrophils. When neutrophils are stimulated with PAF at concentrations greater than 10 nM, a double peak of intracellular calcium mobilization is observed. The second calcium peak observed in PAF-treated neutrophils has already been suggested to come from the production of endogenous leukotriene B4 (LTB4). Here we demonstrate the involvement of endogenous LTB4 production and subsequent activation of the high affinity LTB4 receptor (BLT1) in this second calcium mobilization peak observed after stimulation with PAF. We also show that the second, but not the first peak, could be desensitized by prior exposure to LTB4. Moreover, when neutrophils were pre-treated with pharmacological inhibitors of LTB4 production or with the specific BLT1 antagonist, U75302, PAF-mediated neutrophil degranulation was inhibited by more than 50%. On the other hand, pre-treating neutrophils with the PAF receptor specific antagonist (WEB2086) did not prevent any LTB4-induced degranulation. Also, when human neutrophils were pre-treated with U75302, PAF-mediated chemotaxis was reduced by more than 60%. These data indicate the involvement of BLT1 signaling in PAF-mediated neutrophil activities. PMID- 15789614 TI - Selective translocation of protein kinase c isozymes by PAF in rabbit platelets. AB - The action of platelet activating factor (PAF) on subcellular distribution and activity of protein kinase C (PKC) isoforms in rabbit platelets was analyzed. The results showed an increase of PKC alpha in membrane fraction, concomitantly with a decrease in cytosolic fraction after 5 min PAF treatment, indicating that a translocation of PKC alpha occurred. In addition, PKC zeta was redistributed in a "reverse" form, from the membrane to cytosolic fraction after PAF treatment. PAF induced an increase of PKC alpha activity, whereas a decrease rather than increase in PKC zeta was observed by using immunoprecipitation assays. In addition, some results indicated that PI3 kinase activation was not involved in PAF-induced PKC zeta translocation as occur in several cells and with other agonists. These actions were time- and concentration-dependent, and were inhibited by the treatment with a PAF antagonist. No translocation was observed when the platelets were incubated with lysoPAF, a PAF related compound. The redistribution of PKC isoforms take place through the activation of high specificity PAF binding sites. The pretreatment of the rabbit platelets with staurosporine, a putative inhibitor of PKC, completely blocked the PAF-evoked aggregation without affecting to PAF-evoked shape change and serotonin release. All together, these data could suggest that the specific translocation of PKC isoforms play an important role in the activation of rabbit platelets. PMID- 15789615 TI - On the mechanism of biosynthesis of 19-hydroxyprostaglandins of human seminal fluid and expression of cyclooxygenase-2, PGH 19-hydroxylase (CYP4F8) and microsomal PGE synthase-1 in seminal vesicles and vas deferens. AB - The predominating prostaglandins of human seminal fluid are 19R hydroxyprostaglandins E1 and E2, conceivably formed sequentially by prostaglandin H (PGH) synthase-2, PGH 19-hydroxylase (CYP4F8), and microsomal PGE synthase-1 of seminal vesicles. Our aim was to study this enzyme system. Quantification by real time PCR suggested that the transcripts of PGH synthase-2, CYP4F8, and microsomal PGE synthase-1 were abundant and correlated in seminal vesicles of seven patients (p < 0.05). The three enzymes were detected in seminal vesicles by Western blot analysis, and immunohistological analysis confirmed the localization to the epithelia of seminal vesicles and distal vas deferens. Immunofluorescence analysis showed co-localization of the three enzymes in epithelial cells of seminal vesicles and vas deferens. 19-Hydroxy-PGE compounds were detected by mass spectrometry in the mucosa of distal vas deferens. Recombinant CYP4F8 catalyzes n 2 hydroxylation of PGH1 and PGH2 and n-3 hydroxylation of arachidonic acid. Arachidonic acid was oxidized to 18-hydroxyarachidonic acid and to PGE2 and by microsomes of seminal vesicles in the presence of NADPH and GSH, and to relatively small amounts of 19-hydroxy-PGE2. We conclude that PGH synthase-2, CYP4F8, and PGE synthase-1 likely forms 19-hydroxy-PGE compounds in seminal vesicles and vas deferens, but the catalytic properties of CYP4F8 suggest additional biological functions. Recombinant CYP4F8 was also found to catalyze n 2 hydroxylation of PGI2 and carbaprostacyclin (Km to approximately 40 microM), and n-2 and n-3 hydroxylation of carbocyclic TXA2. PMID- 15789616 TI - Red blood cells: reservoirs of cis- and trans-epoxyeicosatrienoic acids. AB - Epoxyeicosatrienoic acids (EETs) are candidate endothelium-derived hyperpolarizing factors that demonstrate a wide range of biological effects. The presence of both cis- and trans-EETs in rat plasma was identified with HPLC electrospray ionization tandem mass spectrometry in this study. The total EETs in plasma are 38.2 ng/ml with cis-EETs representing 21.4 +/- 0.4 ng/ml and trans EETs 16.8 +/- 0.4 ng/ml. EETs in RBCs were estimated to be 20.2 ng/10(9) RBCs, which corresponds to 200 ng in RBCs contained in 1 ml blood. RBC incubation with 10 mM tert-butyl hydroperoxide resulted in 4.4-fold increase of total cis-EETs (from 9.2 to 40.2 ng/10(9) RBCs) and 5.5-fold increase of total trans-EETs (from 11.0 to 60.8 ng/10(9) RBCs). EETs were released (2 ng/ml) from RBCs after incubation at 37 degrees C for 10 min even after being washed 3 times, indicating that RBCs are reservoirs of plasma EETs. The identification of cis- and trans EETs in RBCs and in plasma as well as their release from RBCs suggest a vasoregulatory role of RBCs in view of their potent vasoactivity. PMID- 15789617 TI - Group X secretory phospholipase A2 can induce arachidonic acid release and eicosanoid production without activation of cytosolic phospholipase A2 alpha. AB - Group X secretory phospholipase A2 (sPLA2-X) and cytosolic phospholipase A2 alpha (cPLA2alpha) are involved in the release of arachidonic acid (AA) from membrane phospholipids linked to the eicosanoid production in various pathological states. Recent studies have indicated the presence of various types of cross-talk between sPLA2s and cPLA2alpha resulting in effective AA release. Here we examined the dependence of sPLA2-X-induced potent AA release on the cPLA2alpha activation by using specific cPLA2alpha or sPLA2 inhibitors as well as cPLA2alpha-deficient mice. We found that Pyrrophenone, a cPLA2alpha-specific inhibitor, did not suppress the sPLA2-X-induced potent AA release and prostaglandin E2 formation in mouse spleen cells. Furthermore, the amount of AA released by sPLA2-X from spleen cells was not significantly altered by cPLA2alpha deficiency. These results suggest that sPLA2-X induces potent AA release without activation of cPLA2a, which might be relevant to eicosanoid production in some pathological states where cPLA2a is not activated. PMID- 15789619 TI - Uterine region-dependent differences in responsiveness to prostaglandins in the non-pregnant porcine myometrium. AB - To clarify the uterine region-dependent distribution of prostanoid receptors, we compared the mechanical responses to selective prostanoid receptor agonists (FP, EP3, DP, EP2) and naturally occurring prostaglandins (PGF2alpha PGE2, PGD2) in longitudinal and circular muscles isolated from three different regions (cornu, corpus and cervix) of the non-pregnant porcine uterus. Expression levels of FP receptor and cyclooxygenase (COX-1 and COX-2) in the respective regions were also examined using RT-PCR and Western blotting. The contractile responses to fluprostenol (an FP agonist) and PGF2alpha in both longitudinal and circular muscles were strongest in the cornu but weak in the corpus and cervix. Expression levels of mRNA and protein of FP receptor were highest in the cornu, consistent with the contractile responses. ONO-AE-248 (an EP3 agonist) caused contraction of both muscle layers, but region-related difference in responsiveness was observed only in the longitudinal muscle. ONO-AE1-259 (an EP2 agonist) inhibited spontaneous contraction of the myometrium, and inhibition was conspicuously stronger in the cervix. PGE2 caused contraction (<100 nM, cornu > corpus = cervix) and inhibition (>300 nM, cornu = corpus < or = cervix) of contractility depending on the concentration in both muscle layers. BW245C (a DP agonist) inhibited the spontaneous contraction, and region-dependent different responsiveness was marked in the longitudinal muscle (cervix = corpus > cornu). COX-1 but not COX-2 was detected in the non-pregnant porcine uterus. Expression level of COX-1 was different in the longitudinal muscle (cornu > corpus = cervix) but the same in the circular muscle. SC-560 inhibited the spontaneous contraction of longitudinal muscles in all regions. The results of the present study indicate that there are region-related heterogeneous distributions of contractile (FP and EP3, cornu > cervix) and relaxant (EP2 and DP, cervix > cornu) prostanoid receptors and COX-1 in the porcine uterus. The results also suggest involvement of endogenous PGs in the regulation of spontaneous uterine contractility. Region related differences in COX-1 and prostanoid receptors might be necessary to produce a gradient of uterine motility decreasing from the cornu to the cervix that manages movement of luminal contents. PMID- 15789618 TI - 5-lipoxygenase expression and activity in aorta from streptozotocin-induced diabetic rats. AB - We previously reported an activation of the 5-lipoxygenase pathway in aorta from streptozotocin-induced diabetic rats. The aim of this study was to investigate whether this activation was associated with an increased expression of 5 lipoxygenase, an increased cysteinyl leukotriene (CysLT) production in response to arachidonic acid or calcium ionophore A23187 and/or a hypersensitivity of the aorta to CysLTs in streptozotocin-induced diabetic rats. In aorta from diabetic and control rats, reverse transcriptase-PCR and western blot analysis with a specific 5-lipoxygenase antibody provided evidence for the presence of 5 lipoxygenase in aorta. However, the expression of 5-lipoxygenase was not significantly different between diabetic and control rats. Challenge by A23187 (10 microM) and arachidonic acid (10 microM and 0.1 mM) with or without A23187 (10 micromol/l) induced a significant increase of CysLT release (measured by enzyme immunoassay) that was in the same range in aorta from control and diabetic rats. In contrast, aortas from diabetic rats showed a greater sensitivity to LTC4 and LTD4 contractile effects. These data suggested that the activation of the 5 lipoxygenase pathway previously reported in streptozotocin-induced diabetic rats could be explained by an augmented sensitivity to CysLTs of the diabetic aorta. PMID- 15789620 TI - Re-evaluation of thin layer chromatography as an alternative method for the quantification of prostaglandins from rat Kupffer cells. AB - In contrast to conventionally used immunoassays, thin layer chromatography (TLC)- by prelabeling of cells with radioactive arachidonic acid (AA)--allows to differentiate between cellularly built and added prostanoids and thus to investigate feedback effects of prostanoids on their own release. PGD2, TXB2 and PGE2 released from zymosan-stimulated Kupffer cells were separated with distinct RF-values, corresponding to those of the pure substances. Quantification of PGD2 and PGE2 gave comparable results with TLC and immunoassays, but measurement in the presence of added prostanoids was only possible with TLC. Moreover TLC was superior to immunoassays in having a longer linear range while being comparably sensitive. Cellularly built TXB2 in its radioactively labeled form was not detectable by TLC. Inhibition of TXB2 release by externally added AA or technical artifacts were excluded, suggesting that the cellular AA-pools used for prostaglandin and thromboxane synthesis differ in their accessibility for added AA. Thus, TLC is a simple, sensitive and precise method for the quantification of cellularly built prostaglandins but not of thromboxane even in the presence of added prostanoids. PMID- 15789621 TI - Direct coupling between arachidonic acid-induced Ca2+ release and Ca2+ entry in HEK293 cells. AB - Arachidonic acid (AA) modulates intracellular Ca2+ signaling via Ca2+ release or/and Ca2+ entry. However, the mechanism underlies either process is unknown; nor is it clear as to whether the two processes are mechanistically linked. By using Fura2/AM, we found that AA induced mobilization of internal Ca2+ store and an increment in Ca2+, Mn2+ and Ba2+ influx in HEK293 cells. The AA-mediated Ca2+ signaling was not due to AA metabolites, and insensitive to capacitative Ca2+ entry inhibitors. Interestingly, isotetrandrine and Gd3+ inhibited both AA induced Ca2+ release and Ca2+ entry in a concentration-dependent manner without affecting Ca2+ discharge caused by carbachol, caffeine, or thapsigargin. Additionally, similar pattern of inhibition was observed with tetracaine treatment. More importantly, the three compounds exhibited almost equal potent inhibition of AA-initiated Ca2+ release as well as Ca2+ influx. Therefore, this study, for the first time, provides evidence for a direct coupling between AA mediated Ca2+ release and Ca2+ entry. PMID- 15789622 TI - Delta12-prostaglandin D2 is a potent and selective CRTH2 receptor agonist and causes activation of human eosinophils and Th2 lymphocytes. AB - Prostaglandin D2 (PGD2) is a lipid mediator produced by mast cells, macrophages and Th2 lymphocytes and has been detected in high concentrations in the airways of asthmatic patients. There are two receptors for PGD2, namely the D prostanoid (DP) receptor and the chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2). The proinflammatory effects of PGD2 leading to recruitment of eosinophils and Th2 lymphocytes into inflamed tissues is thought to be predominantly due to action on CRTH2. Several PGD2 metabolites have been described as potent and selective agonists for CRTH2. In this study we have characterized the activity of delta12-PGD2, a product of PGD2 isomerization by albumin. Delta12-PGD2 induced calcium mobilization in CHO cells expressing human CRTH2 receptor, with efficacy and potency similar to those of PGD2. These effects were blocked by the TP/CRTH2 antagonist ramatroban. delta12-PGD2 bound to CRTH2 receptor with a pKi of 7.63, and a 55-fold selectivity for CRTH2 compared to DP. In Th2 lymphocytes, delta12-PGD2 induced calcium mobilization with high potency and an efficacy similar to that of PGD2. delta12-PGD2 also caused activation of eosinophils as measured by shape change. Taken together, these results show that delta12-PGD2 is a potent and selective agonist for CRTH2 receptor and can cause activation of eosinophils and Th2 lymphocytes. These data also confirm the selective effect of other PGD2 metabolites on CRTH2 and illustrate how the metabolism of PGD2 may influence the pattern of leukocyte infiltration at sites of allergic inflammation. PMID- 15789624 TI - Providers and staff respond to Medicaid managed care: the unintended consequences of reform in New Mexico. AB - In 1997 a new Medicaid managed care (MMC) program called Salud! was implemented by the State of New Mexico. This article serves as an introduction to a special issue of Medical Anthropology Quarterly that assesses the unintended consequences of this reform and its impact on providers and staff who work in clinics, physician offices, and emergency rooms where Medicaid patients are served. MMC fused state and corporate bureaucracies, creating a complex system where enrollment and access was difficult. The special issue focuses on providers' responses to these new structures, including ways in which staff buffer the impact of reform and the role of the discourses of medical necessity and accountability in shaping the way in which MMC functions. PMID- 15789623 TI - A phase I study of 5-fluorouracil, leucovorin, and celecoxib in patients with incurable colorectal cancer. AB - A phase I study of fixed-dose 5-fluorouracil (FU) and leucovorin (LCV), with excalating doses of the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib, was conducted in 16 patients with advanced colorectal adenocarcinoma. At doses typically used to treat arthritis patients (100-200 mg po BID), celecoxib did not increase toxicities expected from the chemotherapy alone. 5-FU and leucovorin did not affect COX-2 inhibition by celecoxib. Preliminary data suggest it is safe to combine celecoxib with standard chemotherapeutic agents, in treatment of patients with colorectal cancer. PMID- 15789625 TI - De facto disentitlement in an information economy: enrollment issues in Medicaid managed care. AB - This article discusses enrollment issues in New Mexico's Medicaid managed care (MMC) system and seeks to illuminate reasons for persistent problems reported by workers and clients. It argues that between 1997 and 2000, the MMC and welfare reforms raised enrollment barriers by complicating and dehumanizing the system, thus "technically disenfranchising" workers and clients. Specifically, the new system increased the need for professional, in-person enrollment assistance precisely when the state decreased its provision of it. Some aspects of the State Child Health Insurance Program (SCHIP) reforms indirectly aggravated those same problems, and though they also significantly lowered barriers in some areas, overall the new system was plagued with preexisting barriers as well as new, unmet needs that produced "de facto disentitlement" to health services. PMID- 15789626 TI - The safety net of the safety net: how federally qualified health centers "subsidize" Medicaid managed care. AB - In this article, I examine the impact of neoliberalism and welfare reform on the delivery of Medicaid, specifically how the advent of Medicaid managed care (MMC) has been wrought with contradictions, placing increased burdens on primary safety net organizations and impacting the many communities they serve. I argue that federally qualified health centers (FQHCs) operate as a primary safety net among safety-net providers, supporting and subsidizing New Mexico's MMC program financially and administratively. By presenting ethnographic data, I will demonstrate how FQHCs pay many of the hidden financial and institutional costs of the shift to managed care. Such findings uncover paradoxes inherent to neoliberal ideologies and privatization, raising questions about the efficacy of a managed care system for Medicaid as well as the future of the health care safety net and access to health care for the diverse populations it serves. PMID- 15789627 TI - Confronting utilization review in New Mexico's Medicaid mental health system: the critical role of "medical necessity". AB - The insertion of managed care into Medicaid services for the mentally ill has created contention about clinical decision making. At the center of this debate is the matter of what constitutes a medical necessity. Employing ethnographic methodology, this study examines utilization review (UR), the context in which decisions concerning the authorization of mental health care services are made. Interviews carried out in the study contrast ideological underpinnings of providers and advocates of the mentally ill, on the one hand, with employees and administrators of managed care institutions, on the other. The result is an exploration into the ways discourses surrounding the mental health care needs of New Mexico's Medicaid population are being constructed and are determining the actual care they receive. PMID- 15789628 TI - Power, blame, and accountability: Medicaid managed care for mental health services in New Mexico. AB - I examine the provision of mental health services to Medicaid recipients in New Mexico to illustrate how managed care accountability models subvert the allocation of responsibility for delivering, monitoring, and improving care for the poor. The downward transfer of responsibility is a phenomenon emergent in this hierarchically organized system. I offer three examples to clarify the implications of accountability discourse. First, I problematize the public private "partnership" between the state and its managed care contractors to illuminate the complexities of exacting state oversight in a medically underserved, rural setting. Second, I discuss the strategic deployment of accountability discourse by members of this partnership to limit use of expensive services by Medicaid recipients. Third, I focus on transportation for Medicaid recipients to show how market triumphalism drives patient care decisions. Providers and patients with the least amount of formal authority and power are typically blamed for system deficiencies. PMID- 15789631 TI - ["Quorum sensing" systems in infective pathology]. AB - Recently the existence of bacterial cell-cell intercommunications, the so called "quorum sensing" systems, has been discovered and studied. The role that these systems play in the infections by Gram-negative and Gram-positive bacteria and the promising therapeutic applications are briefly discussed. PMID- 15789629 TI - Ideologies of aid, practices of power: lessons for Medicaid managed care. AB - The articles in this special issue teach valuable lessons based on what happened in New Mexico with the shift to Medicaid managed care. By reframing these lessons in broader historical and cultural terms with reference to aid programs, we have the opportunity to learn a great deal more about the relationship between poverty, public policy, and ideology. Medicaid as a state and federal aid program in the United States and economic development programs as foreign aid provide useful analogies specifically because they exhibit a variety of parallel patterns. The increasing concatenation of corporate interests with state and nongovernmental interests in aid programs is ultimately producing a less centralized system of power and responsibility. This process of decentralization, however, is not undermining the sources of power behind aid efforts, although it does make the connections between intent, planning, and outcome less direct. Ultimately, the devolution of power produces many unintended consequences for aid policy. But it also reinforces the perspective that aid and the need for it are nonpolitical issues. PMID- 15789630 TI - [Blood doping: 2]. AB - Blood Doping has recently obtained a large diffusion between professional and nonprofessional athletes, in particular for endurance sports it has almost become a necessary way to warrant best performance. Seven years after the publication of our article "Blood Doping", this second installment was born to emphasize the way the biomedical research supplies (often unintentionally) new drugs and new technology to improve athletic performance and, on the other hand, to underline the antidoping strategies. PMID- 15789632 TI - [Management of acute pain in Italy. Economic aspects and quality of life]. AB - OBJECTIVES: This study aims to measure quality of life and treatment costs of patients with acute pain due to osteoarticular diseases;2 to asses associations between pain, quality of life and treatment costs from NHS' perspective. PATIENTS AND METHODS: Observational, multicentric study; patients of both gender were enrolled sequentially from March to July 2002 and observed for 30 days. Type of pain was assessed by a McGill Pain Questionnaire, while degree of severity was measured by VAS (Visual Analogue Scale) ranging from 0 (no pain) to 10 (very severe pain). Quality of life was assessed by EQ-5D (EuroQol/Quality of Life in Europe-5 Dimensions). RESULTS: 188 patients matched by gender and age in to four group: male < or =65 years (51), female < or =65 years (59), male > 65 years (42), female > 65 years (36). Male's age ranged from 36 to 88 years, while female's ranged from 31 to 87 years. Groups differ for type of pain. Cost after enrollment (48.99 Euro) is due for about 60% to diagnostic examinations, while pharmacological therapies account for only 17.64 Euro. Mean score of EQ-5D shows a low level of utility (0.265) that reflects a rather law quality of life. Patient older than 65 years had lower score than patient of the same gender but younger. Differences are significant in the dimension anxiety/depression that afflicts women and older patients. CONCLUSIONS: This study valued utility of patients with non oncological acute pain in Italy and health cost of diagnostic and therapeutic path faced by patients. PMID- 15789633 TI - [Histogenic characterization of the cells forming RA pannus]. AB - OBJECTIVE: This work aims to clarify the histogenesis of the cells forming RA pannus: the pannocytes. METHODS: 15 patients with seropositive RA; 5 controls with post-traumatic knee effusion and 5 with OA knee effusion were included in the study. Synovial tissues and fluids, collected during diagnostic arthrocentesis, were used as a source of cells to be cultured. Viable staining and cytocentrifugation were performed. Cell phenotype was investigated by immunofluorescence assays after different culture periods. Cells were also studied by immunohistochemistry to determine the presence of CD5, CD68:KP-1, CD68:PG-M1, vimentin, cytokeratin, a-SM-Actin. RESULTS: Cells derived from RA samples were sub-divided into two population of lymphocyte-like and macrophage like cells. Phenotypical characteristics of the first one were analysed after 6 days of culture and suggested they were T lymphocytes. The other population could grow in vitro for undefined time resembling the neoplastic-like proliferation previously described for pannocytes. Phenotypic characterization excluded that these cells were lymphocytes, monocyte-macrophages, fibroblasts, myofibroblasts, endothelial cells or keratinocytes. On the contrary, immunohistochemistry demonstrated that 100% of pannocytes were vimentin positive and 75% of these cells expressed also CD68:KP-1. CONCLUSIONS. The results exclude that pannocytes originate from monocyte-macrophages or from fibroblasts, but strongly support the hypothesis that they belong to the family of primitive embryonal connective tissue-forming cells (residue of the primitive mesenchymal tissue). PMID- 15789634 TI - [Asymptomatic hyper-creatine-phosphokinasemia: a veritable disease? Observation in some cases]. AB - Myopathy is a pathology of the muscle function, without clinical and instrumental signs that are involved the central nervous system and the peripheral one. In all pathologies causing a damage of the muscle fibres, a release of enzymes as GOT, GPT, LDH, aldolase, CPK occurs. The most significant enzyme for the muscle damage is CPK. Studies state that an increase of the CPKemia is not always an expression of muscle damages, but there are physiological changes in relation to the age, physical activities and racial variations. By the way, it is necessary to focus our attention on this matter. Authors, starting from the observation of lots of cases under their studies, have described two of them, that up to their careful observation could reveal significant. PMID- 15789635 TI - [Syndrome of overlap: Chronic hepatitis C/autoimmune hepatitis: fact or fancy?]. AB - Non-organ specific autoantibodies are common in patients with chronic hepatitis C, making differential diagnosis difficult between viral, autoimmune forms and hepatitis C/autoimmune hepatitis overlap syndrome. The lack of additional criteria of autoimmunity in most patients leads to the definition of a "false" hepatitis C-autoimmune hepatitis overlap syndrome, while the "true" overlap syndrome occurs in a very few number of subjects. In patients with "false" overlap syndrome the first choice therapy is based on the administration of interferon plus ribavirin. On the contrary, first-line therapy with corticosteroids should be restricted to the "true" hepatitis overlap syndrome with the new therapeutic option of interferon/corticosteroid association. PMID- 15789636 TI - [The thyroid nodule: cytomorphofunctional profile-therapeutic approach]. AB - Analyzing a cohort of 16,400 thyroid nodules, sequentially examined since 1982, the value of preoperative echo-guided fine needle aspiration cytology (FNA) in discriminating benign lesion from malignant ones, has been assessed. Ultrasonography provides a useful support not only to guide the diagnostic FNA methodology, but also monitoring therapeutic procedure: evacuation of cyst, alcoholic sclerotization, laser therapy, effects of treatment on the size of the nodular structure. The correct interpretation of the imaging pictures should be rationally anchored to both clinical criteria and to circumstantial anamnestic analysis, as well as to physical examination, laboratory tests, instrumental systems, cytomorphological patterns, immunohistologic and biomolecular studies. The FNA sampling under ultrasonography guidance offers an absolute diagnostic reliability, and it can be confidently applied for planning surgical strategy. By adopting this safe, non invasive, accurate diagnostic tool, that offers the advantage of eliminating unnecessary operation for benign lesions, the number of operation is strikingly reduced, while it allows to identify an higher surgical frequency of malignancy, yielding a prevalence of about 3% of thyroid nodules. FNA is a very profitable cost-effective diagnostic tool, reducing 20% the cost of care, for the evaluation and treatment of patient with thyroid nodule. Preliminary results on the molecular pattern of thyroid nodules, obtained applying a new methodological system, the Laser Capture Microdissection, are underlined; in the next issue of this journal it will be analytically illustrated the diagnostic role of this innovative procedure that appears very promising in obtaining information on the molecular derangements of a single thyroid cell, even at a precancerous stage; thus a preventive surgical treatment of a thyroid nodule genetically characterized can be predicted. PMID- 15789637 TI - [People die with philosophy]. PMID- 15789638 TI - Another subtle change... PMID- 15789639 TI - Tsunami. PMID- 15789640 TI - New coding and billing opportunities for 2005--Part II. AB - Part I of this series highlighted several changes in preventative care services and additional revisions for pediatricians, orthopedists, endoscopist and surgeons performing transplants and bariatric surgery. Part II addresses several new codes in gynecology, orthopedics, neurosurgery, and ophthalmology. It also includes wound care along with many revisions within chemotherapy for Medicare patients. All new CPT codes should have been activated and deleted codes discontinued effective for dates of service on or after January 1, 2005. From that date forward the patient's date of service must reflect current diagnosis (ICD-9) and procedure/services (CPT and HCPCS) codes. PMID- 15789641 TI - Olecranon stress fracture. PMID- 15789642 TI - Update in rheumatoid arthritis therapy. AB - Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by polyarticular symmetrical arthritis. Inflammatory mediators targeting joint structures produce joint inflammation with pain, functional loss, joint destruction and permanent deformity. Currently, no cure for RA exists but the increasing use of combination therapy and immunomodulatory agents has led to improved quality of life and long-term outlook for many of these patients. While traditionally employed therapies have provided limited disease suppression, advances in our understanding of the molecular pathogenesis of RA have resulted in new therapies targeting very specific components of the inflammatory process. These new treatments have shown very promising results with improved efficacy and an overall decreased toxicity profile. This review provides an overview for practicing clinicians of the current immunosuppressive therapies in RA with an emphasis on newer biological agents regarding their mechanisms of action, efficacy, side effects and monitoring recommendations. Developing therapeutics will be briefly discussed. PMID- 15789644 TI - Patient page. Extreme cold: an outdoor prevention guide for your personal health and safety. PMID- 15789643 TI - Prevention and treatment of the metabolic syndrome in the elderly. AB - The metabolic syndrome is a term used to indicate the presence of a cluster of conditions associated with increased risk for type 2 diabetes, hypertension, coronary artery disease, stroke, and early mortality. A fairly common condition in the elderly, it is caused primarily by physical inactivity and excessive calorie intake and characterized by abdominal obesity, insulin resistance, impaired fasting glucose, dyslipidemia, and prehypertension. Numerous clinical trials have demonstrated that a lifestyle of moderate-intensity, physical activity for 30 minutes a day, most days of the week, combined with weight loss of 5-7%, can reverse individual components of the metabolic syndrome. When lifestyle modifications are insufficient, a multidrug regimen may be necessary to treat different components of the metabolic syndrome. This paper reviews current literature on the metabolic syndrome, including its causes, incidence and approaches for successful treatment. PMID- 15789645 TI - Expect scrutiny of health prices as spending continues upward. PMID- 15789646 TI - Relationships between physiological variables and race performance in French standardbred trotters. AB - The relationships between V4 (the velocity for a blood lactate concentration of 4 mmol/litre) and V200 (the velocity for a heart rate of 200 bpm) and the performance indices of 223 healthy French trotters were analysed. They were divided into four age groups of three, four, five and six years and over, and into three performance groups (good, intermediate and poor performers) defined by the index of trot (ITR), an annual index of performance calculated by the national stud. V4 and V200 were assessed during the performance of a three-step standardised field exercise test, and four performance indexes (ITR, earnings, best time and the number of starts) were calculated for each group. An analysis of variance showed that V4 and V200 were highly correlated with the horses' age and level of performance (P<0.05). The correlation between the physiological variables and ITR was better for the three- and four-year-old horses than for the older horses. PMID- 15789647 TI - Multiple antimicrobial resistant Salmonella enterica serovar Paratyphi B variant Java in cattle: a case report. AB - An epidemiological investigation of a calf rearing premises and a closely associated dairy herd was carried out after the isolation of Salmonella enterica serovar Paratyphi B variant Java phage type 3b variant 2 from clinically diseased calves on the premises. The isolate was resistant to ampicillin, chloramphenicol, streptomycin, sulphonamides, tetracyclines, trimethoprim and cefoperazone. The organism was widespread on the calf unit and was also recovered from the dairy premises, mainly from groups of weaned calves. The investigation was extended to 10 epidemiologically linked farms but no S Java was isolated from any of the 40 to 60 samples collected from each premises. Molecular studies showed that the S Java isolates were genetically most similar to isolates from cases of human disease associated with ornamental fish tanks or feed. Long PCR and resistance gene profiling identified a resistance island which was indistinguishable from the human 'fish tank' strain of S Java and animal and human epidemic strains of S Typhimurium DT104. The isolates were clearly distinguished from multi-resistant S Java strains commonly associated with continental poultry. This is the first report of S Java with this resistance pattern in Great Britain. PMID- 15789648 TI - Efficacy of the treatment of dogs with leishmaniosis with a combination of metronidazole and spiramycin. AB - Twenty-seven dogs infected naturally with Leishmania infantum were used in a randomised controlled trial to compare the clinical and parasitological efficacy of an oral treatment with a combination of metronidazole and spiramycin (13 dogs) with the efficacy of conventional treatment with meglumine antimonate and allopurinol (14 dogs) as controls. In the test group one dog had to be withdrawn from the treatment because it developed pemphigus foliaceus; 10 of the dogs were clinically responsive but none was cured parasitologically. In the control group four dogs were withdrawn from the treatment because of side effects; eight of the dogs were clinically responsive but none was cured parasitologically. The control group showed signs of improvement after an average of 30 days, whereas the test group did not show signs of improvement until after an average of 45 days. PMID- 15789649 TI - Lack of reversion in triclabendazole-resistant Fasciola hepatica. PMID- 15789650 TI - Clinical and ultrasonographic findings in three cows with ruptured gall bladders. PMID- 15789651 TI - Intrathoracic haemangiosarcoma in an ostrich (Struthio camelus). PMID- 15789652 TI - Incidence of Chrysomya bezziana screw-worm myiasis in Saudi Arabia, 1999/2000. PMID- 15789653 TI - Prescribing of proposed POM-VPM products. PMID- 15789654 TI - Restructuring at the IAH. PMID- 15789655 TI - Draft order on the supply of POMs. PMID- 15789656 TI - Restrictions on distance retail selling of POM products. PMID- 15789657 TI - Transplantation of fetal kidney cells: neuroprotection and neuroregeneration. AB - Various trophic factors in the transforming growth factor-beta (TGF-beta) superfamily have been reported to have neuroprotective and neuroregenerative effects. Intracerebral administration of glial cell line-derived neurotrophic factor (GDNF) or bone morphogenetic proteins (BMPs), both members of the TGF-beta family, reduce ischemia- or 6-hydroxydopamine (6-OHDA)-induced injury in adult rat brain. Because BMPs and GDNF are highly expressed in fetal kidney cells, transplantation of fetal kidney tissue could serve as a cellular reservoir for such molecules and protect against neuronal injury induced by ischemia, neurotoxins, or reactive oxygen species. In this review, we discuss preclinical evidence for the efficacy of fetal kidney cell transplantation in neuroprotection and regeneration models. PMID- 15789659 TI - Regulation of trophic factor expression by innervating target regions in intraocular double transplants. AB - Trophic factors have been found to play a significant role both in long-term survival processes and in more rapid and dynamic processes in the brain and spinal cord. However, little is known regarding the regulation of expression of growth factors, and how these proteins interact on a cell-to-cell basis. We have studied protein levels of one growth factor known to affect the noradrenergic innervation of the hippocampal formation, namely brain-derived neurotrophic factor (BDNF). The purpose of the present study was to determine if appropriate innervation or contact between the LC noradrenergic neurons and their target, the hippocampus, affects expression of this growth factor in either brain region. Fetal brain stem tissue, containing the LC, and hippocampal formation were dissected from embryonic day 17 rat fetuses and transplanted together or alone into the anterior chamber of the eye of adult Fisher 344 rats. The tissue was grown together for 6 weeks, after which the animals were sacrificed and ELISAs for BDNF were undertaken. Transplantation to the anterior chamber of the eye increased the expression of BDNF in the hippocampal but not the brain stem tissue, compared with levels observed in fetal and adult rats in vivo. In addition, double grafting with hippocampal tissue more than tripled BDNF levels in brain stem grafts and doubled BDNF levels in the hippocampal portion of double grafts compared with hippocampal single grafts. Triple grafts containing basal forebrain, hippocampus, and brain stem LC tissue increased brain stem and hippocampal BDNF levels even further. Colchicine treatment of LC-hippocampal double grafts gave rise to a significant decrease in hippocampal BDNF levels to levels seen in single hippocampal grafts, while only a partial reduction of BDNF levels was seen in the brain stem portion of the same double grafts treated with colchicine. The findings suggest that an appropriate hippocampal innervation or contact with its target tissues is essential for regulation of BDNF expression in the brain stem, and that retrograde transport of BDNF can occur between double grafted fetal tissues in oculo. PMID- 15789658 TI - Feasibility and safety of autologous myoblast transplantation in patients with ischemic cardiomyopathy. AB - Successful autologous skeletal myoblast transplantation into infarcted myocardium in a variety of animal models has demonstrated improvement in cardiac function. We evaluated the safety and feasibility of transplanting autologous myoblasts into infarcted myocardium of patients undergoing concurrent coronary artery bypass grafting (CABG) or left ventricular assist device implantation (LVAD). In addition, we sought to gain preliminary information on graft survival and any potential improvement of cardiac function. Eighteen patients with a history of ischemic cardiomyopathy participated in a phase I, nonrandomized, multicenter pilot study of autologous skeletal myoblast transplantation concurrent with CABG or LVAD implantation. Twelve patients with a history of previous myocardial infarction (MI) and a left ventricular ejection of less than 30% were enrolled in the CABG arm. In a second arm, six patients underwent LVAD implantation as a bridge to heart transplantation and were required to donate their heart for testing at the time of heart transplant. Myoblasts were successfully transplanted in all patients without any acute injection-related complications or significant long-term unexpected adverse events. Follow-up PET scans showed new areas of viability within the infarct scar in CABG patients. Echocardiography measured an average improvement in left ventricular ejection fraction (LVEF) from 25% to 34%. Histological evaluation in four out of five patients who underwent heart transplantation documented survival and engraftment of the skeletal myoblasts within the infarcted myocardium. These interim results demonstrate survival, feasibility, and safety of autologous myoblast transplantation and suggest that this modality may offer a potential therapeutic treatment for end-stage heart disease. PMID- 15789660 TI - Liver repopulation after hepatocellular transplantation: integration and interaction of transplanted hepatocytes in the host. AB - The mechanisms of donor hepatocyte integration into recipient liver are not fully understood. We investigated mechanisms of both the integration and interaction of transplanted hepatocytes with host liver cells as well as the repopulation of the host organ following intraportal transplantation. Mature hepatocytes were injected into the portal vein of dipeptidylpeptidase IV (DPPIV)-deficient rats pretreated with retrorsine and subjected to 30% partial hepatectomy to ensure selective donor growth. The degree of integration and proliferation was studied by colocalizing transplanted cells (DPPIV positive) with connexin 32, MMP-2, and OX-43 (multilayer immunofluorescence imaging). FACS analysis was established to assess the extent of repopulation quantitatively. Transplanted hepatocytes reached the distal portal spaces and sinusoids within 1 h after injection. A small proportion of cells succeeded in traversing the endothelial barrier through mechanical disruption in both locations. Transplanted hepatocytes lost their membrane-bound gap junctions (connexin 32) during this process. Successful integration of the donor cells required up to 5 days, heralded by gap junction reconstitution and the specific basolateral membrane expression of DPPIV. MMP-2 degraded the extracellular matrix in close proximity to donor cells, providing space for cell division. FACS analysis revealed that more than 37% of the liver was repopulated by cells derived from donors at 2 months after transplantation. Our data demonstrate a high degree of donor cell repopulation of the host organ and provide valuable insight into the specific mechanisms of donor cell integration. Connexin 32 expression in transplanted hepatocytes may serve as an indicator of their effective incorporation and communication within the recipient liver. FACS analysis reveals an accurate method to determine quantitatively the extent of liver repopulation. PMID- 15789661 TI - Monocrotaline, an alternative to retrorsine-based hepatocyte transplantation in rodents. AB - Retrorsine has been used extensively to inhibit proliferation of resident hepatocytes in various transplantation models. Here we report a successful alternative to currently unavailable retrorsine that can be used in cellular transplantation models. Based on structural and molecular similarities, we investigate the use of monocrotaline (MCT) in cell transplantation studies in rodents. In this study, MCT was given to rats intraperitoneally in two injections 2 weeks apart. Two weeks after the final injection, a partial hepatectomy followed by splenic hepatocyte transplantation was performed. The results indicate that MCT, at two doses of 30 mg/kg, highly enhances liver repopulation by donor hepatocytes following partial hepatectomy and produces 15.3 +/- 4.9% liver repopulation within the first 6 weeks following transplantation. Additionally, we tested the effectiveness of MCT in a murine model. Using two injections of 50 mg/kg each, given 2 weeks apart, hepatocyte proliferation in the native liver was inhibited and subsequent oval cell transplants engrafted at 18 +/- 21.3% after 16 weeks posttransplantation. In conclusion, MCT can be used as an effective selective pressure for donor hepatocytes in cell transplantation to the liver in rodents. PMID- 15789662 TI - HVEGF165 increases survival of transplanted hepatocytes within portal radicles: suggested mechanism for early cell engraftment. AB - VEGF is a potent angiogenic factor that promotes hepatocyte growth, increases permeability of blood vessels, and induces vasodilatation, and may accelerate engraftment and function of transplanted hepatocytes. The aim was to study the effect of VEGF on early hepatocyte engraftment. Thirty-two Lewis syngeneic female rats underwent 70% partial hepatectomy. Eighteen received 240 ng VEGF165 and 14 received saline for control. Thereafter, intrasplenic transplantation of 10(7) male hepatocytes was done. Semiquantitative analysis of PCR product of the SRY region of the Y-chromosome was performed. Paraffin-embedded sections were stained for H&E and for PCNA immunostaining. By PCR, male hepatocytes were identified in 8 livers out of 14 VEGF-treated rats at 24-48 h, compared with only 1 liver out of 8 controls. Transplanted cells were seen within portal vessels radicles in 7 out of 14 VEGF-treated rats for as long as 48 h posttransplantation, compared with only one control liver at 24 h. There was no histological sign of cell injury to transplanted or adjacent cells. Two weeks after transplantation male transplanted cells were identified in two out of four rats treated with hVEGF165 and in one out of six rats treated with saline. No transplanted cells were detected within portal tracts 14 days after transplantation. hVEGF165 enhances the presence of transplanted hepatocytes within portal vessels after transplantation. We suggest an additional mechanism for cell engraftment, whereby transplanted hepatocytes first stick to each other in the portal radicles. Later they become included in the liver parenchyma as groups of organized cells in a process stimulated by VEGF. PMID- 15789663 TI - Short-term culture with the caspase inhibitor z-VAD.fmk reduces beta cell apoptosis in transplanted islets and improves the metabolic outcome of the graft. AB - In the initial days after transplantation islets are particularly vulnerable and show increased apoptosis and necrosis. We have studied the effects of caspase inhibition on this early beta cell death in syngeneically transplanted islets. Streptozotocin-diabetic C57BL/6 mice were transplanted with 150 syngeneic islets, an insufficient mass to restore normoglycemia, preincubated with or without the pan-caspase inhibitor z-VAD. fmk 2 h before transplantation. Beta cell apoptosis was increased in control islets on day 3 after transplantation (0.28 +/- 0.02%) compared with freshly isolated islets (0.08 +/- 0.02%, p < 0.001), and was partially reduced in transplanted islets preincubated with z-VAD.fmk 200 microM (0.14 +/- 0.02%, p = 0.003) or with z-VAD.fmk 500 microM (0.17 +/- 0.01%, p = 0.012), but not with a lower z-VAD.fmk (100 microM) concentration. Diabetic mice transplanted with islets preincubated with z-VAD.fmk 500 microM showed an improved metabolic evolution compared with control and z-VAD.fmk 200 microM groups. The z-VAD.fmk 500 microM group showed an overall lower blood glucose after transplantation (p = 0.02), and at the end of the study blood glucose values were reduced compared with transplantation day (15.7 +/- 3.6 vs. 32.5 +/- 0.5 mmol/L, p = 0.001). In contrast, blood glucose was not significantly changed in control and z-VAD.fmk 200 microM groups. Four weeks after transplantation beta cell mass was higher in z-VAD.fmk 500 microM group (0.15 +/- 0.02 mg) than in the control group (0.10 +/- 0.02 mg) (p = 0.043). In summary, the treatment of freshly isolated islets with the caspase inhibitor z-VAD.fmk reduced the subsequent apoptosis of the islets once they were transplanted and improved the outcome of the graft. PMID- 15789665 TI - Physician assistants and humanitarian assistance. PMID- 15789664 TI - Central necrosis in isolated hypoxic human pancreatic islets: evidence for postisolation ischemia. AB - A variety of explanations have been provided to elucidate the requirement of the large islet mass that is essential for a successful treatment of patients with type I diabetes by intrahepatic transplantation. The purpose of this study was to investigate islet cell survival under the effect of prolonged hypoxia and/or nutrient withdrawal, which mimics posttransplantation environment of transplanted islets in the liver. We studied the influence of 24 h of hypoxia (1% O2) in intact isolated human and rat islets as well as the effect of combined oxygen/nutrient deprivation in a mouse insulinoma cell line (MIN6). In intact human islets, 24 h of hypoxia led to central necrosis combined with apoptotic features such as nuclear pyknosis and DNA fragmentation. In the course of hypoxic treatment, ultrastructural analysis demonstrated a gradual transition from an apoptotic to a necrotic morphology particularly pronounced in central areas of large islets. In MIN6 cells, on the other hand, hypoxia led to a twofold (p < 0.01) increase in caspase-3 activity, an indicator of apoptosis, but not to necrosis, as determined by release of lactate dehydrogenase (LDH). Only in combination with nutrient/serum deprivation was a marked increase in LDH release observed (sixfold vs. control, p < 0.01). We therefore conclude that, similar to MIN6 cells, central necrosis in isolated hypoxic islets is the result of the combined effects of hypoxia and nutrient/serum deprivation, most likely due to limited diffusion. Provided that transplanted islets undergo a similar fate as shown in our in vitro study, future emphasis will require the development of strategies that protect the islet graft from early cell death and accelerate the revascularization process. PMID- 15789666 TI - A tale of two coxibs. PMID- 15789667 TI - Evaluating acute abdominal pain in adults. PMID- 15789668 TI - The metabolic syndrome: modify root causes, treat risk factors. PMID- 15789669 TI - Preparing for disasters: what should you know, and when should you know it? PMID- 15789670 TI - Making a bad situation worse. PMID- 15789671 TI - Case of the month. Subdural hematoma. PMID- 15789673 TI - Workplace aggression destroyed this nurse's career. PMID- 15789672 TI - Beyond the bracelet. PMID- 15789674 TI - On the frontline in Iraq: these nurses are our heroes. PMID- 15789675 TI - On the frontline in Iraq: these nurses are our heroes. PMID- 15789676 TI - On the frontline in Iraq: these nurses are our heroes. PMID- 15789677 TI - This reader offers advice on breaking an ampule cleanly. PMID- 15789678 TI - Nurses address end-of-life care professionally and personally. PMID- 15789679 TI - Nurses address end-of-life care professionally and personally. PMID- 15789680 TI - Nurses address end-of-life care professionally and personally. PMID- 15789681 TI - Healthy eating or just another diet myth? PMID- 15789682 TI - Anyone can perform a task--nurses can assess and evaluate. PMID- 15789683 TI - Question of the month. What do you wish you had been taught in nursing school- but weren't? PMID- 15789684 TI - Ethics in action. PMID- 15789685 TI - Cosmetic surgery: is it really right for your patient? PMID- 15789686 TI - How to manage an acute upper GI bleed. PMID- 15789687 TI - The silent thief. PMID- 15789688 TI - Would you suspect this genetic disorder? PMID- 15789689 TI - VAP prevention. The latest guidelines. PMID- 15789690 TI - A guide for those returning to the bedside. PMID- 15789692 TI - Why is quality care still elusive? PMID- 15789691 TI - Statistical methodology of the National Immunization Survey, 1994-2002. AB - OBJECTIVES: Since 1994 the National Immunization Survey (NIS) has monitored progress toward the Healthy People 2000 and 2010 vaccination goals. The NIS collects data in two phases: first, a random-digit-dialing (RDD) telephone survey to identify households with children 19-35 months old and, second, a mail survey to vaccination providers to obtain vaccination histories used to estimate vaccination coverage rates. This report reviews the methodologies used in the 1994-2002 NIS to obtain official estimates of vaccination coverage and describes the methodology used for the first three topical modules of the NIS. METHODS: From 1994 to 1997 the NIS used a variation of a two-phase estimator to compensate for missing provider-reported vaccination histories. Between 1998 and 2001 a weighting-class estimator was used. In 2002 and thereafter the weighting-class approach was refined to account for households that do not have telephones and for unvaccinated children. To collect data on immunization-related topics, the NIS sample was randomized among three topical modules: health insurance and ability to pay for vaccinations (HIM); parental knowledge and experiences about vaccinations (PKM); and daycare attendance, breastfeeding practices, and participation in the Special Supplemental Nutrition Program for Women, Infants, and Children (DCM). RESULTS: In 2001 among children with completed RDD interviews, 0.3 percent were entirely unvaccinated. Together, the new nontelephone adjustment and the refinement for unvaccinated children yielded revised estimates that were within 1.5 percentage points of the original estimates obtained using the 1998-2001 methodology. Over the six quarters during which the first three topical modules were fielded (from mid-2001 through 2002), 21,163 children were randomized to the HIM, 3576 to the PKM, and 3511 to the DCM. PMID- 15789693 TI - The rewards of rehabilitation nursing: a story of gratitude. PMID- 15789694 TI - The ethics of using cybernetics and cyborg technologies: what every rehabilitation nurse should know. AB - Cybernetics and cyborg technologies are rapidly developing in the field of biotechnology. Such developments have yielded a wide variety of devices and prosthetics that have promoted the quality of life for many individuals with physical limitations and generally have been applauded by society and the rehabilitation field. However, such rapid developments have given rise to multiple ethical concerns. Understanding these ethical concerns and the implications they have for rehabilitation nurses is imperative. While the potential benefits of advances in technology are great for those with disabilities and chronic conditions, ethicists suggest that skepticism must be balanced with the zeal that often accompanies cutting-edge developments. As Hook notes, "We must show not a fear of technology, but a courageous control of technology, and refuse to let technology control us" (2002, p. 67). PMID- 15789695 TI - How do we prevent UTI in people who perform intermittent catheterization? PMID- 15789696 TI - Health-related quality of life: is it a good indicator of function post THR? AB - The purpose of this study was to explore the impact of health-related quality of life (HRQOL) measured with the Short Form Health Survey (SF-36) on Functional Recovery Status (physical and psychosocial recovery status) at base-line, 2 months, 6 months, and 12 months following total hip replacement (THR). A secondary analysis was performed using data gathered from a sample of 271 older adults post THR. Four empirically based hypothesized models were tested. None of the models fit the data, with each having significant chi2 values and chi2 /df ratios greater than 3. Different dimensions of HRQOL at baseline, 2, 6, and 12 months were related to physical recovery status, and none of the 8 dimensions of the SF-36 was significantly related to psychosocial recovery status. Overall, the results of this study do not support the hypothesis that HRQOL, as measured by the SF-36, comprehensively explains functional recovery status following THR. Clinically, these findings may be applicable to individuals at risk for poor recovery. They also may prompt practitioners to consider alternative factors that influence psychosocial recovery. PMID- 15789697 TI - Psychometric evaluation of selected pain intensity scales for use with cognitively impaired and cognitively intact older adults. AB - The purpose of this study was to determine the reliability and validity of selected pain intensity scales such as the Faces Pain Scale (FPS), the Verbal Descriptor Scale (VDS), the Numeric Rating Scale (NRS), and the Iowa Pain Thermometer (IPT) to assess pain in cognitively impaired older adults. A descriptive correlational design was used, and a convenience sample of 66 volunteers age 60 and older residing in assisted living facilities in the South was recruited for this study. The sample included 22 (33%) men and 44 (67%) women, with a mean age of 76. Ninety-eight percent (65) of the sample comprised Caucasian participants, with the exception of 1 African-American man. Seventy percent (47) completed high school and/or college. The mean Mini Mental State Exam (MMSE) score was 16, with a range of 1 to 29. Eighty-five percent scored 24 or lower, indicating some degree of cognitive impairment. The remaining 15% were cognitively intact. All but one participant could use each scale to rate their pain. Concurrent validity of the VDS, NRS, and IPT was supported with Spearman rank correlation coefficients ranging from .78 to .86 in the cognitively impaired group. The FPS, however, demonstrated weak correlations with other scales when used with the impaired group, ranging from .48 to .53. In the cognitively intact group, strong correlations ranging from .96 to .97 were found among all of the scales. Test-retest reliability at a 2-week interval was acceptable in the cognitively intact group (Spearman rank correlations ranged from .67 to .85) and unacceptable for most scales in the cognitively impaired group (correlations ranged from .26 to .67). When asked about scale preference, both the cognitively impaired and the intact groups preferred the IPT and the VDS. This study revealed that cognitive impairment did not inhibit participants' ability to use a variety of pain intensity scales, but the stability issue must be considered. PMID- 15789698 TI - A theoretical model for preventing osteoarthritis-related disability. AB - Osteoarthritis (OA) affects many aspects of life for affected individuals. Effective interventions to prevent and restore function must be based upon an understanding of what contributes to OA and its associated disabilities. A hypothetical OA disability model built upon the previous work of Nagi (1991), Verbrugge and Jette (1994), the International Classification of Functioning of World Health Organization (World Health Organization, 2001), and other scientific findings is proposed. The model includes a main disease pathway, which describes the sequence of events from OA-associated impairments to disabilities. Contextual factors influencing the process include individual characteristics, psychological state, coping style, comorbidities, social support, and physical environment. The model provides a useful conceptual framework for understanding the OA disability process from a biopsychosocial perspective and for guiding rehabilitation nursing interventions in OA care. PMID- 15789699 TI - Psychological hardiness in women with paraplegia. AB - A revised model of psychological hardiness suggests that transformational coping, positive health behaviors, and activistic social support may be qualitative indicators of hardiness and an expression of the synergistic presence of commitment, control, and challenge. Hardiness indicators correlate with health rather than illness, and have an indirect effect on the stress-illness relationship through health. Stress and illness also correlate with health. A secondary qualitative analysis of the transcripts of eight women with paraplegia revealed three major themes related to spinal cord injury (SCI): physiological aspects of SCI, associated with the traumatic event and its consequences; psychological aspects of SCI, associated with problem-solving, choices, and challenges; and social aspects of SCI, associated with personal and professional relationships. PMID- 15789700 TI - Analytical evaluation of chromatic dispersion in photonic crystal fibers. AB - We present a two-dimensional modal approach for the evaluation, in an analytical manner, of chromatic dispersion in any kind of optical fiber. It combines an iterative Fourier technique to compute the propagation constant at any fixed wavelength and an analytical procedure to calculate its derivatives. The proposed formulation takes into account the effective anisotropy of the interfaces and allows us to deal with microstructured fibers, in general, and specifically with realistic photonic crystal fibers (PCFs), including arbitrary spatial refractive index distributions of dispersive and absorbing materials. This fast and accurate numerical technique is extremely useful for both analysis and design. We show some results of analysis of PCFs with high anisotropy, and we also describe PCFs with new dispersive properties. PMID- 15789701 TI - High-gain Tm-doped fluoride fiber amplifier. AB - A diode-pumped Tm-doped fiber-optic amplifier that has a small-signal gain of >30 dB at 1870 nm is reported. Output pulses of up to 3-W peak power at a 1-60-kHz repetition rate can be generated by amplification of 20-40-ns laser diode pulses of up to 2-mW launched peak power. The output signal quality, i.e., the ratio of the output pulse energy and the total amplified spontaneous emission (ASE) output energy between two pulses, depends on the relative propagation direction of pump and signal and can be dramatically increased by choice of the correct propagation scheme. In the optimum pump geometry the pulse energy can be raised to as much as 20 times the ASE energy. This is the first report to the author's knowledge of fiber-optic amplification of short diode laser pulses near 1.9 microm with high repetition rates in Tm-doped fibers. PMID- 15789702 TI - Single-frequency, single-mode, plane-polarized ytterbium-doped fiber master oscillator power amplifier source with 264 W of output power. AB - We present a single-frequency, single-mode, plane-polarized ytterbium-doped all fiber master oscillator power amplifier source at 1060 nm generating 264 W of continuous-wave output power. The final-stage amplifier operated with a high gain of 19 dB and a high conversion efficiency of 68%. There was no evidence of rollover from stimulated Brillouin scattering even at the highest output power, and the maximum output was limited only by the available pump power. PMID- 15789703 TI - Permanent refractive-index modification in germanium-doped optical fibers by use of red light. AB - We observed photoinduced birefringence in elliptical-core optical fibers by using a continuous-wave krypton-ion laser. We induced the birefringence by injecting 20 mW of 647-nm or 50 mW of 676-nm light into the fiber at 45 degrees to the slow axis. The rate of change of the refractive index was found to be proportional to the square of the average power. Polarization mode couplers written into the fibers have been stable for more than 2 years and can be erased by use of light polarized perpendicularly to the original writing beam. PMID- 15789704 TI - Design of air-guiding honeycomb photonic bandgap fiber. AB - We introduce a design procedure for hollow-core photonic bandgap (PBG) fiber with a cladding made of air-silica honeycomb photonic crystal (PC). It is found that air-guiding can be realized in both fundamental and secondary PBG regions of the cladding PC. Dispersion and radiation loss of the fundamental mode for two types of fiber structure are theoretically calculated. The fibers show promising waveguiding ability. PMID- 15789705 TI - Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy. AB - We have developed a digital holographic microscope (DHM), in a transmission mode, especially dedicated to the quantitative visualization of phase objects such as living cells. The method is based on an original numerical algorithm presented in detail elsewhere [Cuche et al., Appl. Opt. 38, 6994 (1999)]. DHM images of living cells in culture are shown for what is to our knowledge the first time. They represent the distribution of the optical path length over the cell, which has been measured with subwavelength accuracy. These DHM images are compared with those obtained by use of the widely used phase contrast and Nomarski differential interference contrast techniques. PMID- 15789706 TI - Object characterization with refractometric digital Fourier holography. AB - We demonstrate a digital holographic method in which two different substances in a blend are discerned. The method requires only one set of exposures and one reconstruction in the plane of focus. The phase is unwrapped by Flynn's discontinuity algorithm to produce an image of the variation of the optical distance of the illuminating wave. Objects with indices of refraction that are higher and lower than the mounting liquid are detected as regions in which the phase is increased and decreased, respectively. We also present a method for calculating the volume distribution of substrates in a sample. The method is experimentally demonstrated with crystals of NaCl and KCl. PMID- 15789707 TI - Chaos-assisted tunneling in dielectric microcavities. AB - We demonstrate that the lifetimes and emission patterns of the optical modes in generic (asymmetric) microresonators are strongly affected by the phenomenon of chaos-assisted tunneling and develop a theory of the effect. PMID- 15789708 TI - Structure and scaling of helical modes of light. AB - Modes of light that contain topological defects such as screw dislocations can be focused into optical traps with interesting and useful properties. The way in which the intensity distribution within helical modes of light varies with topological charge is discussed, and new scaling predictions for their radial profiles that are consistent with experimental observations are introduced. PMID- 15789709 TI - InAs/GaAs quantum-dot saturable absorbers for diode-pumped passively Q-switched Nd-doped 1.3-microm lasers. AB - A low-loss semiconductor saturable absorber based on InAs/GaAs quantum dots was developed for Q switching of a diode-pumped Nd-doped laser operating at 1.3 microm. With an InAs/GaAs quantum-dot saturable absorber, a diode-pumped Nd:YVO4 laser at 1342 nm was achieved. With an incident pump power of 2.2 W, an average output power of 360 mW with a Q-switched pulse width of 90 ns at a pulse repetition rate of 770 kHz was obtained. PMID- 15789710 TI - Improved photoluminescent properties in one-dimensional LaPO4:Eu3+ nanowires. AB - The photoluminescent properties of one-dimensional LaPO4:Eu nanowires were studied in contrast with the corresponding zero-dimensional nanoparticles, micrometer particles, and micrometer rods. The results indicate that in nanowires Eu3+ ions occupied two different symmetry sites, unlike in nanoparticles and micrometer particles. Furthermore, the radiative transition rate of 5D1 - sigmaJ 7FJ or 5D0 - sigmaJ 7FJ and the internal luminescent quantum efficiency of Eu3+ in the nanowires showed obvious improvement over that in the other powders. PMID- 15789711 TI - Self-seeded multiwavelength Brillouin-erbium fiber laser. AB - We propose and demonstrate a self-seeded multiwavelength Brillouin-erbium fiber laser with an internally self-excited Brillouin pump, which is achieved by incorporation of a length of single-mode fiber together with a Sagnac loop mirror into a fiber ring cavity. In this simple scheme the Brillouin pump is self excited in the fiber ring cavity and then used to seed the Brillouin multiwavelength comb in the single-mode fiber. Stable generation of more than 120 Brillouin Stokes wavelengths with relatively uniform amplitudes is demonstrated with this scheme. It is also shown that such a self-seeded Brillouin laser has good stability and repeatability. PMID- 15789712 TI - Real data results with wavelength-diverse blind deconvolution. AB - Multiframe blind deconvolution is extended to incorporate simultaneous image acquisition at multiple wavelengths (wavelength diversity). The assumption of common path-length errors across the diversity channels allows for a parallel deconvolution procedure that exploits this coupling. No assumptions about variations in the object's intensity distribution at different wavelengths are required. The method is described and initial results for real images collected with a bench-scale imaging system are presented. PMID- 15789713 TI - Algorithm for near-field reconstruction based on radial-shearing interferometry. AB - A new iterative algorithm to be used to precisely reconstruct near-field distribution from an interferogram of a laser output generated by a cyclic radial shearing interferometer is proposed. First, by use of a window function around the zero-frequency part of the Fourier transform of the interferogram and calculation of the inverse Fourier transform of the zero-frequency part, we obtain the background intensity distribution of the interferogram. Then, according to the iterative algorithm, the near-field distribution of the laser output is precisely reconstructed from the background intensity distribution obtained in the first step. A numerical simulation and an actual experiment of the near-field reconstruction of the laser output with arbitrary amplitude distribution are implemented successfully. PMID- 15789714 TI - Molecularly sensitive optical coherence tomography. AB - Molecular contrast in optical coherence tomography (OCT) is demonstrated by use of coherent anti-Stokes Raman scattering (CARS) for molecular sensitivity. Femtosecond laser pulses are focused into a sample by use of a low-numerical aperture lens to generate CARS photons, and the backreflected CARS signal is interferometrically measured. With the chemical selectivity provided by CARS and the advanced imaging capabilities of OCT, this technique may be useful for molecular contrast imaging in biological tissues. CARS can be generated and interferometrically measured over at least 600 microm of the depth of field of a low-numerical-aperture objective. PMID- 15789715 TI - Mode transformer for miniaturized optical circuits. AB - A novel mode transformer was fabricated that transforms a modal area by a factor of 100. Using the mode transformer improves the efficiency of mode transformation by an order of magnitude compared with that when no mode transformer is used. With this mode transformer, input-output coupling of miniaturized, on-chip integrated optical circuits to external optical fibers is achieved with low loss. The mode transformer's design, fabricated in silicon, is scalable to virtually any waveguide size, facilitating continuous miniaturization in silicon optoelectronics. PMID- 15789716 TI - Photoinduced grating formation in azo-dye-labeled phospholipid thin films by 244 nm light. AB - Holographic gratings are recorded in azo-dye nitrobenzoxazole-labeled phospholipid thin films by use of 244-nm UV light. The gratings continue to grow for more than 1 h, even after the recording light is removed. The diffraction efficiency of these gratings shows extreme sensitivity to humidity and can increase reversibly by 2 orders of magnitude in air that is saturated with water vapor. This effect is related to the unique characteristics of phospholipid molecules that undergo hydration-dependent structural reorganization and self assembly. PMID- 15789717 TI - Optical stimulation of neural tissue in vivo. AB - For more than a century, the traditional method of stimulating neural activity has been based on electrical methods, and it remains the gold standard to date. We report a technological breakthrough in neural activation in which low-level, pulsed infrared laser light is used to elicit compound nerve and muscle potentials in mammalian peripheral nerve in vivo. Optically induced neural action potentials are spatially precise, artifact free, and damage free and are generated by use of energies well below tissue ablation threshold. Thus optical stimulation presents a simple yet novel approach to contact-free in vivo neural activation that has major implications for clinical neurosurgery, basic neurophysiology, and neuroscience. PMID- 15789718 TI - Deeply penetrating photoacoustic tomography in biological tissues enhanced with an optical contrast agent. AB - Photoacoustic tomography (PAT) in a circular scanning configuration was developed to image deeply embedded optical heterogeneity in biological tissues. While the optical penetration was maximized with near-infrared laser pulses of 800-nm wavelength, the optical contrast was enhanced by Indocyanine Green (ICG) dye whose absorption peak matched the laser wavelength. This optimized PAT was able to image objects embedded at depths of as much as 5.2 cm, 6.2 times the 1/e optical penetration depth, in chicken breast muscle at a resolution of <780 microm and a sensitivity of <7 pmol of ICG in blood. The resolution was found to deteriorate slowly with increasing imaging depth. The effects of detection bandwidth on the quality of images acquired simultaneously by four different ultrasonic transducers are described. PMID- 15789719 TI - Nanolayer characterization through wavelength multiplexing of a microsphere resonator. AB - We optically characterize nanolayer (<150 nm) formation in situ on a silica microsphere in an aqueous environment by simultaneously following the shifts of whispering-gallery modes at two wavelengths. This approach was inspired by layer perturbation theory, which indicates that these two measurements can be used to determine independently both the thickness and the optical dielectric constant. The theory is verified for extreme cases and used to characterize a biophysically relevant hydrogel nanolayer with an extremely small excess refractive index of 0.0012. PMID- 15789720 TI - Expanding the bandwidth of slow-light photonic devices based on coupled resonators. AB - It is shown theoretically that canceling third-order dispersion can substantially increase the useful bandwidths of linear and nonlinear optical devices based on slow propagation of light. Cancellation on both global and local scales can be achieved by combination of the ring-based coupled resonator lines and all-pass optical filters. PMID- 15789721 TI - Nonparaxial dark solitons in optical Kerr media. AB - We show that the nonlinear equation that describes nonparaxial Kerr propagation, together with the already reported bright-soliton solutions, admits of (1 + 1)D dark-soliton solutions. Unlike their paraxial counterparts, dark solitons can be excited only if their asymptotic normalized intensity u2infinity is below 3/7; their width becomes constant when u2infinity approaches this value. PMID- 15789722 TI - Soliton-induced waveguides in an organic photorefractive glass. AB - We demonstrate optical waveguiding of a probe beam at 980 nm by a soliton beam at 780 nm in an organic photorefractive monolithic glass. Both planar and circular waveguides induced by one- and two-dimensional spatial solitons formed as a result of orientationally enhanced photorefractive nonlinearity are produced in the organic glass. Possibilities for increasing the speed of waveguide formation are discussed. PMID- 15789723 TI - Reflective-type configuration for nearly phase-matching cerenkov second-harmonic generation in a nonlinear-optical polymer waveguide. AB - A new technique for achieving efficient Cerenkov-type second-harmonic generation (SHG) in a nonlinear-optical (NLO) polymer waveguide is presented. The configuration, which can prevent the losses of light caused by relatively long distance propagation and the multiple reflections that appear in the conventional Cerenkov technique, exhibits ease of fabrication and compactness. We experimentally observed a conversion efficiency of 1.6% W(-1) cm(-1), which to our knowledge is the highest value reported for Cerenkov SHG in polymer, by tuning both the thickness and the refractive index of the polymer film close to phase matching between a guided fundamental wave and a guided harmonic wave. The experimental results agreed well with the theoretical prediction. PMID- 15789724 TI - Resonant radiation and collapse of ultrashort pulses in planar waveguides. AB - We study the spatiotemporal dynamics of ultrashort pulses close to a point of zero group-velocity dispersion in planar waveguides with focusing nonlinearities. We find that the process of pulse collapse enhances the emission of so-called resonant radiation, providing an efficient mechanism of energy transfer from solitonic to dispersive waves and leading to suppression of the collapse. PMID- 15789725 TI - 10-GHz dispersion-managed soliton fiber-optical parametric oscillator using regenerative mode locking. AB - We demonstrate a regeneratively mode-locked fiber-optical parametric oscillator that utilizes intracavity dispersion compensation to generate pulses at a 10-GHz repetition rate in both soliton and nonsoliton regimes. At the threshold pump power the generated pulses are close to fundamental solitons. At higher pump powers we found a significant deviation of the pulses from the sech2 shape. In addition, the use of an ultralow-jitter self-starting pump-pulse source in a regenerative feedback loop allows for a significant reduction of the signal's timing jitter and amplitude noise. PMID- 15789726 TI - Stability properties of multiwavelength, incoherent, dissipative spatial solitons. AB - We have investigated the interaction between two dissipative spatial solitons of different frequencies in periodically patterned semiconductor optical amplifiers. The experimental results are in good agreement with the theory. Simulations suggest that multiwavelength interactions do not produce stable bound solitons unless the system's modeling equations are completely symmetric. PMID- 15789727 TI - Polarization-dependent supercontinuum generation from light filaments in air. AB - We investigate polarization-dependent properties of the supercontinuum emission generated from filaments produced by intense femtosecond laser pulses propagating through air over a long distance. The conversion efficiency from the 800-nm fundamental to white light is observed to be higher for circular polarization than for linear polarization when the laser intensity exceeds the threshold of the breakdown of air. PMID- 15789728 TI - Fabrication of photonic crystals in ZnS-doped glass. AB - Three-dimensional photonic crystals were fabricated in a ZnS-doped silicate glass by use of a femtosecond pulsed laser. Woodpile structures consisting of 36 layers were produced by focusing with a 100x (1.35-numerical-aperture) objective. Attenuations of approximately 3-8 dB arising from a photonic bandgap were observed in the visible and the near-infrared regions. PMID- 15789729 TI - Ince-Gaussian series representation of the two-dimensional fractional Fourier transform. AB - We introduce the Ince-Gaussian series representation of the two-dimensional fractional Fourier transform in elliptical coordinates. A physical interpretation is provided in terms of field propagation in quadratic graded-index media whose eigenmodes in elliptical coordinates are derived for the first time to our knowledge. The kernel of the new series representation is expressed in terms of Ince-Gaussian functions. The equivalence among the Hermite-Gaussian, Laguerre Gaussian, and Ince-Gaussian series representations is verified by establishing the relation among the three definitions. PMID- 15789730 TI - Designing optical-fiber modulators by using magnetic fluids. AB - To reduce interface loss between optical fibers and devices in telecommunication systems, the development of an optical-fiber-based device that can be fused directly with fibers is important. A novel optical modulator consisting of a bare fiber core surrounded by magnetic fluids instead of by a SiO2 cladding layer is proposed. Applying a magnetic field raises the refractive index of the magnetic fluid. Thus we can control the occurrence of total reflection at the interface between the fiber core and the magnetic fluid when light propagates along the fiber. As a result, the intensity of the outgoing light is modulated by variation in field strength. Details of the design, fabrication, and working properties of such a modulator are presented. PMID- 15789731 TI - Experimental demonstration of waveguide-coupled round-cornered octagonal microresonators in silicon nitride. AB - We report laterally waveguide-coupled octagonal microresonators with designed round resonator corners in silicon nitride. We demonstrate nearly single-mode add drop filter characteristics, when 50-microm-size round-cornered octagonal microresonators are used, with an optimum coupling efficiency exceeding 96%, a finesse of approximately 30, and a Q of approximately 6400. Our experiments also reveal two unforeseen phenomena in certain microresonator shapes: a pronounced drop in coupling efficiency and resonance notches in the drop spectrum. These device characteristics hint at the recently postulated multiple orbit interference in waveguide-coupled polygonal microresonators. PMID- 15789732 TI - Highly tunable photonic crystal filter for the terahertz range. AB - By use of an incipient ferroelectric, SrTiO3, as a defect material inserted into a periodic structure of alternating layers of quartz and high-permittivity ceramic, thermal tuning of a single defect mode over the entire lowest forbidden band was obtained. The tunability of this compact structure reached 60%. PMID- 15789733 TI - Microcavity confinement based on an anomalous zero group-velocity waveguide mode. AB - We propose and demonstrate a mechanism for small-modal-volume high-Q cavities based on an anomalous uniform waveguide mode that has zero group velocity at a nonzero wave vector. In a short piece of a uniform waveguide with a specially designed cross section, light is confined longitudinally by small group-velocity propagation and transversely by a reflective cladding. The quality factor Q is greatly enhanced by the small group velocity for a set of cavity lengths that are separated by approximately pi/k0, where k0 is the longitudinal wave vector for which the group velocity is zero. PMID- 15789734 TI - Single-scattering spectroscopy for extremely dense colloidal suspensions by use of a low-coherence interferometer. AB - Single-scattering spectroscopy by use of a low-coherence interferometer is introduced to measure the power spectra of light scattered from extremely dense colloidal suspensions. The power spectrum of a heterodyne component can be obtained by subtraction of the power spectrum of a homodyne component from the measured power spectrum. The heterodyne power spectrum for light scattered from the medium is shown to coincide with the single-scattering spectrum to a depth of up to a few times the mean-free path length. Therefore single-scattering spectroscopy is newly proposed as a means by which to analyze the characteristics of extremely dense colloidal suspensions. PMID- 15789735 TI - Photoinduced phase transition in VO2 nanocrystals: ultrafast control of surface plasmon resonance. AB - We study the ultrafast insulator-to-metal transition in nanoparticles of VO2, obtained by ion implantation and self-assembly in silica. The nonmagnetic, strongly correlated compound VO2 undergoes a reversible phase transition, which can be photoinduced on an ultrafast time scale. In the nanoparticles, prompt formation of the metallic state results in the appearance of surface-plasmon resonance. We achieve large, ultrafast enhancement of optical absorption in the near-infrared spectral region that encompasses the wavelength range for optical fiber communications. One can further tailor the response of the nanoparticles by controlling their shape. PMID- 15789736 TI - Generation of synchronized femtosecond and picosecond pulses in a dual-wavelength femtosecond Ti:sapphire laser. AB - We obtain synchronized 45-fs and 0.848-ps pulses by achieving cross-mode locking in a double-cavity dual-wavelength femtosecond Ti:sapphire laser. Autocorrelation and cross correlation show that the femtosecond and picosecond pulses are well synchronized, with a timing jitter of 41 fs. Cross-phase modulation dominates the processes of cross-mode locking and synchronization. PMID- 15789737 TI - Attosecond pulse extreme-ultraviolet photoionization in a two-color laser field. AB - Attosecond-pulse extreme-ultraviolet (XUV) photoionization in a two-color laser field is investigated. Attosecond pulse trains with different numbers of pulses are examined, and their strong dependence on photoelectronic spectra is found. Single-color driving-laser-field-assisted attosecond XUV photoionization cannot determine the number of attosecond pulses from the photoelectronic energy spectrum that are detected orthogonally to the beam direction and the electric field vector of the linearly polarized laser field. A two-color-field-assisted XUV photoionization scheme is proposed for directly determining the number of attosecond pulses from a spectrum detected orthogonally. PMID- 15789738 TI - Multimillijoule chirped parametric amplification of few-cycle pulses. AB - The concept of optical parametric chirped-pulse amplification is applied to attain pulses with energies up to 8 mJ and a bandwidth of more than 100 THz. Stretched broadband seed pulses from a Ti:sapphire oscillator are amplified in a multistage noncollinear type I phase-matched beta-barium borate parametric amplifier by use of an independent picosecond laser with lock-to-clock repetition rate synchronization. Partial compression of amplified pulses is demonstrated down to a 10-fs duration with a down-chirped pulse stretcher and a nearly lossless compressor comprising bulk material and positive-dispersion chirped mirrors. PMID- 15789739 TI - Demonstration of a HeNe/CH4-based optical molecular clock. AB - We implement a simple optical clock based on the F2(2) [P(7), v3] optical transition in methane. A single femtosecond laser's frequency comb undergoes difference frequency generation to provide an IR comb at 3.39 microm with a null carrier-envelope offset. This IR comb provides a phase-coherent link between the 88-THz optical reference and the rf repetition rate. Comparison of the repetition rate signal with a second femtosecond comb stabilized to molecular iodine shows an instability of 1.2 x 10(-13) at 1 s, limited by microwave detection of the repetition rates. The single-sideband phase noise of the microwave signal, normalized to a carrier frequency of 1 GHz, is below -93 dBc/Hz at 1-Hz offset. PMID- 15789740 TI - A stereoselective asymmetric synthesis of antibiotic (-)-fumagillol using claisen rearrangement and intramolecular ester enolate alkylation as key steps. AB - (-)-Fumagillol (1), a hydrolysis product of fumagillin, has been synthesized by several group from commercially available 1,2:5,6-di-O-isopropylidene-alpha-D allofuranose in a highly stereoselective manner. Chiral centers on C5 and C6 came from D-allofuranose and the asymmetric center on C4 was accomplished by 1,3 chirality transfer using the Claisen rearrangement on a chiral allyl alcohol. Chirality, which is necessary on an epoxide consisting of the spiro-ring system, was diastereoselectively constructed by the well-known reaction, intramolecular ester enolate alkylation (IEEA), which showed that this reaction can be applied to the alpha-alkoxy ester system. The epoxide on the side chain was regioselectively introduced by the difference between the number of substituents on the vinyl groups. This accomplishment proved that IEEA can be a useful tool for the synthesis of complex molecules. PMID- 15789741 TI - Design and synthesis of novel antidiabetic agents. AB - The synthesis and structure-activity relationships of a novel series of substituted quercetins that activates peroxisome proliferator-activated receptor gamma (PPARgamma) are reported. The PPARgamma agonistic activity of the most potent compound in this series is comparable to that of the thiazolidinedione based antidiabetic drugs currently in clinical use. PMID- 15789742 TI - Diastereoselective synthesis of polysubstituted pyrrolidinone as a key intermediate for the anticancer agents by palladium(II)-catalyzed carboxylation. AB - Palladium(II)-catalyzed carboxylation of chiral olefins 6a-d has been examined under various conditions. In the weak basic condition (K2CO3), 7a-d were obtained in good yields. Alternatively, in the strong basic condition, pyrrolidinones 8a-d were obtained resulting in excellent yields and with high diastereoselectivity. PMID- 15789743 TI - Inhibition of calmodulin-dependent protein kinase II by cyclic and linear peptide alkaloids from Zizyphus species. AB - The effects of sedative peptide alkaloids from Zizyphus species on calmodulin- dependent protein kinase II were investigated. Protein kinase II activity was assayed on the basis of its ability to activate tryptophan 5-monooxygenase as its substrate in the presence of calmodulin. All thirteen alkaloids tested were stronger inhibitors than chlorpromazine (IC50, 98 microM) on calmodulin-dependent protein kinase II. Among them, the most potent inhibitor was daechuine S27 (IC50 2.95 microM), which was stronger than pimozide (IC50, 15.0 microM). PMID- 15789744 TI - The inhibition of diacylglycerol acyltransferase by terpenoids from Youngia koidzumiana. AB - The EtOAc extract of Youngia koidzumiana significantly inhibited the diacylglycerol acyltransferase (DGAT) from rat liver microsomes. Bioactivity guided fractionation led to the isolation of nine compounds, the structures of which were established using physicochemical and spectral data. Of the isolated compounds, oleanolic acid (2), methyl ursolate (7) and corosolic aicd (8) inhibited DGAT, with IC50 values of 31.7, 26.4, and 44.3 microM, respectively. However, sesquiterpenoids showed only weak inhibitory effects toward DGAT. PMID- 15789745 TI - Inhibitory activity of isorhamnetin from Persicaria thunbergii on Farnesyl Protein Transferase. AB - The methanolic extract of the aerial parts of Persicaria thunbergii was found to show inhibitory activity on Farnesyl Protein Transferase (FPTase). Bioassay guided fractionation of the methanolic extract resulted in the isolation of isorhamnetin, as an inhibitor on FPTase. This compound inhibited FPTase activity in a dose-dependent manner, and the IC50 value of isorhamnetin was 37.5 microM. PMID- 15789746 TI - Butyrylcholinesterase inhibitory guaianolides from Amberboa ramosa. AB - Phytochemical investigation of the whole plant of Amberboa ramosa led to the isolation of six sesquiterpene lactones which could be identified as 8alpha hydroxy-11beta-methyl-1alphaH, 5alphaH, 6betaH, 7alphaH, 11alphaH-guai-10(14), 4(15)-dien-6, 12-olide(1), 3beta, 8alpha-dihydroxy-11alpha-methyl-1alphaH, 5alphaH, 6betaH, 7alphaH, 11betaH-guai-10(14), 4 (15)-dien-6, 12-olide (2), 3beta, 4alpha, 8alpha-trihydroxy-4beta-(hydroxymethyl)-1alphaH, 5alphaH, 6betaH, 7alphaH-guai-10(14), 11(13)-dien-6, 12-olide (3), 3beta, 4alpha, 8alpha trihydroxy-4beta-(chloromethyl)-1alphaH, 5alphaH, 6betaH, 7alphaH-guai 10(14),11(13)-dien-6, 12-olide(4), 3beta, 4alpha, dihydroxy-4beta-(hydroxymethyl) 1alphaH, 5alphaH, 6betaH, 7alphaH-guai-10(14),11(13)-dien-6, 12-olide(5), 3beta, 4alpha-dihydroxy-4beta-(chloromethyl)-8alpha-(4-hydroxymethacrylate)-1alphaH, 5alphaH, 6betaH, 7alphaH-guai-10(14),11 (13)-dien-6,12-olide (6) by spectroscopic methods. All of them showed inhibitory potential against butyrylcholinesterase. PMID- 15789747 TI - Nitric oxide and prostaglandin E2 synthesis inhibitory activities of diarylheptanoids from the barks of Alnus japonica steudel. AB - Nine known diarylheptanoids (1-9) isolated from the barks of Alnus japonica were evaluated for their inhibitory activities on nitric oxide (NO) and prostagrandin E2 (COX-2) production in interferon-gamma (INF-gamma) and lipopolysaccharide (LPS)-activated RAW 264.7 cells in vitro. The NO and COX-2 levels were moderately reduced by the addition of compounds (1-9). Among these compounds, compounds 6 and 8 inhibited NO production in a dose dependent manner with an IC50 of 16.7 and 27.2 microg/mL, respectively (positive control, L-NMMA; 22.8 microg/mL), and compounds 6, 7, 8, and 9 reduced the COX-2 level in a dose dependent manner with an IC50 of 20.7, 25.7, 25.0, and 27.3 microg/mL, respectively (positive control, indomethacin; 26.2 microg/mL). An analysis of the structural activity relationship among these diarylheptanoids suggests that the presence of a keto enol group in the heptane moiety or a caffeoyl group in the aromatic ring were important for the efficacy on the inhibitory activities of NO and COX-2 production. PMID- 15789748 TI - In vitro inhibitory effect of triterpenoidal saponins from Platycodi Radix on pancreatic lipase. AB - In the process of investigating anti-obesity effect of Platycodi Radix, we found that aqueous extract of Platycodi Radix might inhibit intestinal absorption of dietary fat by inhibiting pancreatic lipase (PL) activity. In order to clarify the anti-obesity mechanism of Platycodi Radix, activity-guided isolation was performed to find active components. The total saponin fraction of Platycodi Radix appeared to have a potent inhibitory activity against the hydrolysis of triolein emulsified with phosphatidycholine by pancreatic lipase in vitro. Based on these results, further purification of active components yielded 10 known triterpenoidal saponins, among these compounds, platycodin A, C, D, and deapioplatycodin D exhibited significant inhibitory effects on PL at the concentration of 500 microg/mL with 3.3, 5.2, 34.8, and 11.67% pancreatic lipase activity vs control, respectively. Platycodin D was found to inhibit the PL activity in a dose-dependent manner. Therefore, the anti-obesity effect of Platycodi Radix might be due to the inhibition of pancreatic lipase by its saponins. PMID- 15789749 TI - Anti-estrogenic activity of lignans from Acanthopanax chiisanensis root. AB - Anti-estrogenic activity of (-)-sesamin (1), helioxanthin (2), savinin (3), taiwanin C (4), and 3-(3,4-dimethoxybenzyl)-2-(3,4 methylenedioxybenzyl)butyrolactone (5) isolated from the root of Acanthopanax chiisanensis was tested using Ishikawa cells. Among them, compound 3 exhibited anti-estrogenic activity (IC50 = 4.86 microM). PMID- 15789750 TI - Monoamine oxidase inhibitory components from the roots of Sophora flavescens. AB - In our search for monoamine oxidase (MAO) inhibitors from natural resources, we found that the methanol extract of the roots of Sophora flavescens showed an inhibitory effect on mouse brain monoamine oxidase (MAO). Bioactivity-guided isolation of the extract yielded two known flavonoids, formononetin (1) and kushenol F (2), as active compounds along with three inactive compounds, oxymatrine (3), trifolirhizin (4), and beta-sitosterol (5). Formononetin (1) and kushenol F (2) showed significant inhibitory effects on MAO in a dose-dependent manner with IC50 values of 13.2 and 69.9 microM, respectively. Formononetin (1) showed a slightly more potent inhibitory effect against MAO-B (IC50: 11.0 microM) than MAO-A (IC50: 21.2 microM). Kushenol F (2) also preferentially inhibited the MAO-B activity than MAO-A activity with the IC50 values of 63.1 and 103.7 microM, respectively. PMID- 15789751 TI - The isolation and antioxidative effects of vitexin from Acer palmatum. AB - Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental factors are critical players in cellular damage and aging. In order to develop a new anti-photoaging agent, this work focused on the antioxidant effects of the extract of tinged autumnal leaves of Acer palmatum. One compound was isolated from an ethyl acetate soluble fraction of the A. palmatum extract using silica gel column chromatography. The chemical structure was identified as apigenin-8-C-beta-D-glucopyranoside, more commonly known as vitexin, by spectral analysis including LC-MS, FT-IR, UV, 1H-, and 13C-NMR. The biological activities of vitexin were investigated for the potential application of its anti-aging effects in the cosmetic field. Vitexin inhibited superoxide radicals by about 70% at a concentration of 100 microg/mL and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals by about 60% at a concentration of 100 microg/mL. Intracellular ROS scavenging activity was indicated by increases in dichlorofluorescein (DCF) fluorescence upon exposure to UVB 20 mJ/cm2 in cultured human dermal fibroblasts (HDFs) after the treatment of vitexin. The results show that oxidation of 5-(6 )chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) is inhibited by vitexin effectively and that vitexin has a potent free radical scavenging activity in UVB-irradiated HDFs. In ROS imaging using a confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our findings suggest that vitexin can be effectively used for the prevention of UV induced adverse skin reactions such as free radical production and skin cell damage. PMID- 15789752 TI - Scoparone from Artemisia capillaris inhibits the release of inflammatory mediators in RAW 264.7 cells upon stimulation cells by interferon-gamma Plus LPS. AB - Scoparone is a major component of the shoot of Artemisia capillaris (Compositae), which has been used for the treatment of hepatitis and biliary tract infection in oriental countries. In the present study we observed that, scorparone exhibited no cytotoxic effect in unstimulated macrophages, but reduced the release of nitric oxide (NO) and prostaglandin E2 (PGE2) upon stimulation by IFN-gamma/LPS or LPS. The inhibitory effects were found to be in conjuction with the suppression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX 2) in IFN-gamma/LPS stimulated RAW 264.7 cells. Moreover, scoparone also attenuated the production of tumor necrosis factor (TNF)-alpha, interleukin (IL) 1beta and IL-6 in LPS-stimulated RAW264.7 cells. These results suggest that scoparone decreases the production of the inflammatory mediators such as NO and PGE2 in macrophages by inhibiting iNOS and COX-2 expression. PMID- 15789753 TI - Antinociceptive effect of nicotine in various pain models in the mouse. AB - The antinociceptive effect of nicotine administered intracereboventricularly (i.c.v.) or intrathecally (i.t) in several pain models was examined in the present study. We found that i.t. treatment with nicotine (from 5 to 20 g) dose dependently blocked pain behavior revealed during the second phase, but not during the first phase in the formalin test. In addition, i.c.v. treatment with nicotine (from 0.1 to 10 microg) dose-dependently attenuated pain behavior revealed during both the first and second phases. In addition to the formalin test, nicotine administered i.c.v. or i.t. attenuated acetic acid-induced writhing response. Furthermore, i.c.v. or i.t. administration of nicotine did not cause licking, scratching and biting responses induced by substance P, glutamate, TNF-alpha (100 pg), IL-1beta (100 pg) and INF-gamma (100 pg) injectied i.t. The antinociception induced by supraspinally-administered nicotine appears to be more effective than that resulting from spinally administered nicotine. Our results suggest that nicotine administration induces antinociception by acting on the central nervous system and has differing antinociceptive profiles according to the various pain models. PMID- 15789755 TI - Effect of low adapted temperature and medium composition on growth and erythropoietin (EPO) production by Chinese hamster ovary cells. AB - Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over expresses the first two enzymes of the urea cycle, carbamoyl phosphate synthetase I (CPS I) and ornithine transcarbamoylase (OTC). When supplements were added to the medium at 33 degrees C, the growth of IBE and CO5 cells increased by 27% and 26%, respectively and the maximum yield of EPO was increased by 40% in both cell lines. The absolute EPO concentration in the CO5 cells was always 55-60% higher than in the IBE cells. In addition, when the two cell lines were continuously cultured with supplements at 33 degrees C until their growth rates approached those at 37 degrees C, the growth rates of both IBE and CO5 cells increased by 54% and their maximum EPO levels increased by up to 73% and 56%, respectively. Therefore, the growth and EPO expression levels of CO5 cells increased 2.2-fold and 2.6-fold, respectively, compared to those of the IBE cells. These results indicate that adaptation to lower temperature as well as medium supplementation could be important for improving cell growth and EPO production. PMID- 15789754 TI - Synthetic wogonin derivatives suppress lipopolysaccharide-induced nitric oxide production and hydrogen peroxide-induced cytotoxicity. AB - Wogonin (5,7-dihydroxy-8-methoxyflavone) has been reported to exhibit a variety of biological properties including anti-inflammatory and neuroprotective functions. In this study, biological activities of diverse synthetic wogonin derivatives have been evaluated in two experimental cell culture models. Inhibitory activities of wogonin derivatives on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in BV2 microglial cells and on hydrogen peroxide (H2O2)-induced neuronal cell death in SH-SY5Y human neuroblastoma were examined. Wogonin derivatives such as WS2 and WS3 showed more potent suppressive activities on LPS-induced NO production and H2O2-induced cytotoxicity than wogonin itself. In addition, thiol substitution played a minor role in enhancing the activities of the derivatives. These findings may contribute to the development of novel anti-inflammatory and neuroprotective agents derived from wogonin. PMID- 15789757 TI - Identification of inducible genes during mast cell differentiation. AB - Mast cells play an important role in allergic inflammation by releasing their bioactive mediators. The function of mast cells is enhanced by stimulation because of the induction of specific genes and their products. While many inducible genes have been elucidated, we speculated that a significant number of genes remain to be identified. Thus, we applied differential display (dd) PCR to establish a profile of the induced genes in bone marrow-derived mast cells (BMMCs) after they were co-cultured with 3T3 fibroblasts. To date, 150 cDNA fragments from the connective-type mast cells (CTMCs) were amplified. Among them, thirty cDNA fragments were reamplified for cloning and sequencing. The ddPCR strategy revealed that serine proteases were the most abundant genes among the sequenced clones induced during the maturation. Additionally, unknown genes from the co-culture of BMMCs with 3T3 fibroblasts were identified. We confirmed their induction in the CTMCs by Northern blot analysis and RT-PCR. Characterization of these induced genes during the maturation processes will provide insight into the functions of mast cells. PMID- 15789756 TI - Formalin pretreatment attenuates tail-flick inhibition induced by beta-endorphin administered intracerebroventricularly or intrathecally in mice. AB - We examined the effect of the subcutaneous (s.c.) pretreatment of formalin into both hind paws of mice on the antinociception induced by the intracerebroventricularly (i.c.v.) or intrathecally (i.t.) administration of beta endorphin using the tail-flick test. Pretreatment with formalin (5%) for 5 h had no affect on the i.c.v. administered beta-endorphin-induced tail-flick response. However, pretreatment with formalin for 40 h attenuated the tail-flick inhibition induced by i.c.v. administered beta-endorphin. This antinociceptive tolerance to i.c.v. beta-endorphin continued up to 1 week, but to a lesser extent. Pretreatment with formalin for 5 and 40 h significantly reduced the i.t. beta endorphin-induced inhibition of the tail-flick response, which continued up to 1 week. The s.c. formalin treatment increased the hypothalamic pro-opiomelanocortin (POMC) mRNA level at 2 h, but this returned to the basal level after 40 h. Our results suggest that the increase in the POMC mRNA level in the hypothalamus appears to be involved in the supraspinal or spinal beta-endorphin-induced antinociceptive tolerance in formalin-induced inflammatory pain. PMID- 15789758 TI - Benzodiazepine system is involved in hyperalgesia in rats induced by the exposure to extremely low frequency magnetic fields. AB - Many reports demonstrate that extremely low frequency magnetic fields (ELF MFs, 60 Hz) may be involved in hyperalgesia. In a previous investigation, we suggested that MFs may produce hyperalgesia and such a response may be regulated by the benzodiazepine system. In order to further confirm this effect of MFs, we used diazepam and/or flumazenil with MFs exposure. When testing the pain threshold of rats using hot plate tests, MFs or diazepam (0.5 microg, i.c.v.; a benzodiazepine receptor agonist) induced hyperalgesic effects with the reduction of latency. These effects were blocked by a pretreatment of flumazenil (1.5 mg/kg, i.p.; a benzodiazepine receptor antagonist). When the rats were exposed simultaneously to MFs and diazepam, the latency tended to decrease without statistical significance. The induction of hyperalgesia by co-exposure to MFs and diazepam was also blocked by flumazenil. However, the pretreatment of GABA receptor antagonists such as bicuculline (0.1 microg, i.c.v.; a GABA(A) antagonist) or phaclofen (10 microg, i.c.v.; a GABA(B) antagonist) did not antagonize the hyperalgesic effect of MFs. These results suggest that the benzodiazepine system may be involved in MFs-induced hyperalgesia. PMID- 15789759 TI - Transdermal delivery of piroxicam using microemulsions. AB - To improve the skin permeability of piroxicam, a new oil-in-water microemulsion containing 0.5% piroxicam was developed. Among various oils investigated for their suitability as an oil phase for the microemulsion system, oleic acid showed both excellent solubility and skin permeation enhancing effect for piroxicam. Microemulsion existence ranges were identified through the construction of the pseudo-ternary phase diagram. The effect of the content of oleic acid and the ratio of the surfactant/cosurfactant on skin permeation of piroxicam were evaluated with excised rat skins. The optimum formulation with the highest skin permeation rate (47.14 microg/cm2/h) consisted of 0.5% piroxicam, 10% oleic acid, 60% Labrasol/ethanol (1:5) and water. PMID- 15789760 TI - [Determination of amylose content in transgenic progenies containing an antisense waxy gene and identifying of pure lines in indica rice]. AB - Through genetic transformation mediated by Agrobacterium tumefaciens, an antisense waxy gene was introduced into Longtefu B line, a male sterile maintainer line in indica (Oryza sativa L.). Thirty transgenic plants showed integration of antisense waxy gene into the genome as determined by PCR assay, and twenty eight were confirmed by Southern blotting. T1 seeds from twenty one transgenic plants showed a marked decrease of amylose content, ranging from 3% 13% less than control, and seeds from some transgenic lines exhibited typical waxy phenotype. Six transgenic lines were selected to examine the amylose content in different generations. In the T4 generation, two homozygous lines, Long 3-1-1 1 and Long 5-8-2-1, were selected, with amylose content of 15.9% and 8.4% respectively. The amylose content in these lines is in consistent with the decrease of the accumulation of Wx protein as determined by SDS-PAGE electrophoresis. Cross and subsequent backcross of Long 3-1-1-1 and Long 5-8-2-1 with the Longtefu mare sterile line were performed to determine the changes of amylose content in F1 and B1F1 seeds. The results showed an average amlyose content of 21.4% in F1 seeds with Long 3-1-1-1 as a parent, while only 13.6% with Long 5-8-2-1 as a parent. In addition, the average amylose contents in B1F1 seeds were 17.1% and 9.3% respectively. Our results indicated that during the fertility transfer in the male sterile line, stable transgenic lines with medium or low contents of amylose had direct effects on amylose content in F1 seeds. PMID- 15789761 TI - [Enhancement of plantlet regenerative efficiency of watermelon in vitro]. AB - The purpose of this study was to enhance shoot regenerative efficiency of watermelon by optimizing culture parameters. Cotyledons at 4-day seedlings were explanted and cultured on MS medium supplemented with different combinations of hormones. In "Yixuan" cultivar, the plantlet regenerative frequency and shoots produced by per explant was as low as 10% and 0.4 respectively at all combinations tested when distal parts of cotyledons were inoculated. Whereas, maximum regenerative frequency of 100 percentage and 10.3 shoots regenerated from per explant were observed on MS medium supplemented with 5 mg/L BA and 0.1 mg/L IAA using basal parts of cotyledons as explants. Likewise, in "Jingxinmuben" cultivar, the highest regenerative frequency of 100 percentage and 6.9 shoots regenerated from per explant were obtained when proximal parts of cotyledons were cultured on MS medium containing 2 mg/L BA and 0.5 mg/L IAA. In the present study, adventitious buds were initiated after only 4 d in culture and intact plantlets were obtained in 40 d culture. This will facilitate quicker gene transformation and polyploidy induction of watermelon in vitro. PMID- 15789762 TI - [Pattern of plasma sex steroid hormone levels during the breeding season of male and female skink: Eumeces chinensis]. AB - Changes in gonadal activity and plasma sex steroid hormone levels in male and female Eumece chinensis during the breeding season were described. The results showed that: The vitellogensis of follicles of female Eumeces chinensis needed the stimulation of 17beta-estradiol (E2). As ovary masses reached peak values between late April and mid-May, E2 levels rose to the top value by late March, and then sharply declined but went up again before preovulation; The physiological functions of plasma progesterone (P) consisted in its oviductal egg retention, embryo development, and eggshell formation. P levels fluctuated near the basic value between mid-March and late April. In mid-May, with the onset of ovulation, plasma P levels rose rapidly, reached peak value by late May and declined sharply after ovulation. Plasma E2 levels declined as plasma P levels rose, showing an inverse relationship between them; In males, plasma Testosterone (T) levels were closely correlated with the maintenance of spermatogenesis activities, male and male combat, sexual display, and mating. Plasma T levels tended to rise after the termination of hibernation, and reached peak value by mid-April. After mid-May, with the testis aggressing, plasma T levels gradually went down and reached bottom value by late June. PMID- 15789763 TI - [The investigation of Syn and NPY expression in brain tissues of diabetic model rat induced by streptozotocin]. AB - The synaptophysin (Syn) and neuropeptide Y (NPY) expressions in brain tissue of diabetic model rats were investigated. 20 adult Sprague-Dawley male rats were randomly divided into two groups. 10 rats were injected with streptozotocin to induce diabetes, which was identified by blood & urinary sugar level; and the others were injected with sodium citrate buffer as the control. After 4 weeks' diabetes appearance, the learning and memory ability of each rat in both group were tested by Y-Maze. Afterward freeze sections from the frontal cortex and hippocampus were made and stained with Nissl and Syn, NPY Immunohistochemistry staining. It was observed that diabetic rats showed more errors in Y-Maze test than those control rats. Both the number of neurons and the light densities of Syn and NPY immunostaining reaction in the frontal cortex and hippocampus of brain tissue sections significantly decreased in the diabetic model rats. It is suggested that the decrease in neuron number and Syn, NPY expression in frontal cortex and hippocampus may be one of the factors leading to diabetic dementia. PMID- 15789764 TI - [The evolutionary traits of mitocondrial ND4-ND4L in Drosophila melanogaster species group]. AB - The sequences of the mitochondrial ND4-ND4L were determined for all the 33 extant taxaes of the Drosophila melanogaster species group, so evolutionary traits of ND4 gene sequences were analyzed. We found a strong transition bias in closed related species and saturation of transitional substitutions in distant related species. Moreover, we calculated the diverge time of eight subgroup according to the Tamura-Nei distance between D. melanogaster and D. yakuba. Our result suggested that ananassae subgroup branched off firstly, and montium, melanogaster, ficsphila, eugracilis, elegans, suzukii-takahashii branched off sequentially. PMID- 15789765 TI - [Distribution and toxicity of cadmium in Hydrilla verticillata (L.f.) Royle]. AB - Aquatic plants are known to accumulate and bioconcentrate heavy metals. In this study, aquatic vascular plant Hydrilla verticillata (L.f.) Royle was cultivated in water containing elevated concentrations of cadmium (up to 10 mg/L) for 7 d, the accumulation, subcellular distribution, ultrastructural localization, chemical form, toxic effects on mineral nutrient absorption of cadmium, photosynthesis rate and respiration rate were studied. It was found that H. verticillata fronds was able to accumulate cadmium, the bioconcentration factor was 193-307; subcellular fraction analysis revealed that cadmium major accumulated in cell wall (61.66%-52.00%) with decrease trends, the content of Cd enhanced in soluble fraction and remained stable in organelles, with the augment of pollutant concentration. The Cd levels occurred in different parts of leaf cell with the following sequence: cell wall > soluble fraction > organelles. Ultrastructural localization of cadmium with sulfide-silver method showed that Cd appeared in cell wall, chloroplast, nucleus and vacuole. Sequential extraction indicated that the ratio of different cadmium chemical form was different markedly, of which NaCl extractable Cd was predominated as compared with the other 5 forms; which could be seen in the following order: F(NaCl) > F(HAc) >F(Water) > F(Ethanol) > F(HCl) > F(Residue). The mineral nutrient absorption was also affected by cadmium stress, it increased the absorption of Ca, Mn, Cu, and Fe; but reduced that of P and K. Cd had a strong inhibitive effect on photosynthesis rate and respiration rate. The results suggested that the toxic symptoms of plant showed an evident correlation between dose and effect; the ultrastructural damage was closely related to the distribution of Cd. The conclusion could be reached that the death of plant was resulted from destruction of structure foundation of physiological function, unbalance of ion equilibrium and disorder of physiological metabolism. PMID- 15789766 TI - [Establishment of high-yield suspension cell line of Curcuma zedoaria (Berg.) Rosc and study on the volatile oil synthesis-controlling with precursors]. AB - The condition for high-yield suspension cell line and the precursors of volatile oil synthesis of Curcuma zedoaria (Berg.) Rosc were studied. The results showed that the light yellow particle callus was suitable for establishment of the high yield suspension cell line. The optimum conditions for cell growth were MS medium added 15-30 g/L glucose and 15-30 g/L sucrose (1:1) as carbon source, the total concentration of 80 mmol/L nitrogen source combined NH4+ with NO3- (1:3), hormones of 3.0-5.0 mg/L 6-BA, 1.0 mg/L 2,4-D and dark culture after 10-15 days light culture. The 229 g/L cell (FW) and 2.11% content of volatile oil were obtained in vitro. The addition of precursors of calcium pantothenate, ammonium acetate and potassium acetate during the middle period of the cell suspension culture enhanced the volatile oil content respectively, and ammonium acetate was most effective among them. The highest yield of volatile oil obtained was 3.11% and 8.27 g/L respectively , which was 1.25 and 1.2 times of the control group. PMID- 15789767 TI - [Isolation and prelimary characterization of a kind of rat liver potential progenitor cells small round hepatic cells]. AB - Liver undergoes profound regeneration usually after hepatic damage. It has been shown in recent study that two kinds of liver stem cells, which are mainly oval cells (OVCs) and small hepatocytes, are involved in the process of liver regeneration as they differentiated into premature liver cells. However, the origination of oval cells as well as its differentiation property is not quite understood. In this study, we isolated a novel potential liver progenitor cells, namely small round cells (SRCs). The cellular features of the cells such as morphological appearance, surface marker, growth curve, and differentiation induced by DMSO were analyzed. SRCs and OVCs were obtained by using discontinuous digestions and isopyknic centrifugation, respectively. With non-radioactive measurement of MTT, the cell growth was assayed. Meanwhile, SRCs were stained with antibodies against CK19, ALB and AFP antibody to characterize their tissue specificities. The results showed that SRCs appeared in round-shaped, but irregular in size. The nuclei of SRCs were relative small with rich nucleolus visible. SRCs were semi-floated during primary culture. Even cultured in F12:DMEM (1:1) mixed-medium supplemented with 15% fetal calf serum, viability of the cells could merely be expanded over 7 days. SRCs were positively stained by CK19, ALB and AFP within initial 3 days. After DMSO stimulation, SRCs were not only morphologically changed into OVCs, but also expressed those markers identical to OVCs. It was suggested that SRCs could be considered as an potential candidate of liver progenitors which were related to OVCs. PMID- 15789768 TI - [Analysis of genetic diversity of 14 species of Cymbidium based on RAPDs and AFLPs]. AB - RAPDs and AFLPs were used to investigate the genetic variation of 14 species of Cymbidium. A great diversity was shown by 12 random primers and three primer combinations. UPGMA cluster analysis showed that the results from the RAPD and AFLP analysis were similar and the genetic distances were closest between C. ensifolium and C. sinense, C. kanran and C. faberi var. omeiense, C. mastersii and C. eburneum respectively. PMID- 15789769 TI - [Suppression of apple polyphenol oxidase by double-stranded RNA (RNAi)]. AB - Antisense and sense gene fragments (710 base pairs) of apple polyphenol oxidase (APPO) gene were obtained by RT-PCR amplification, using the total RNAs isolated from ripen apple fruit as the template. These two fragments were ligated with a 1000 bp spacer, YYT (crtW+crtY fusion) gene, which is relative to carotenoid synthesization in subcocci. The full-length 2446 bp-target gene was then inserted into plant binary vector pYPX145 to generate the recombinant plasmid pYF7704, which carried the expression unit, of APPO dsRNA. pYF7704 was transformed to apple (Malus x domestica) var. Red Fuji via agrobacterium tumefaciens mediated leaf disc transformation. With the selection of Karamycin and GUS detection assays, transgenic shoots of A PPO dsRNA were obtained. The results of FQ-RT-PCR indicated that APPO mRNA level was suppressed to 91.69% in transgenic shoots compared to wide shoots. The data suggested that dsRNAi technology on apple polyphenol oxidase is feasible to be utilized in transgenic shoots. PMID- 15789770 TI - [Function of auxin-binding protein gene during cucumber fruit development]. AB - A cDNA fragment that encodes auxin-binding protein 1 was amplified by, reverse polymerase chain reaction from ovary of cucumber. Its expression signals were weak in the ovary of 1 d before anthesis, while got strong in 2, 4 and 6 d after pollination. Among the unpollinated ovary of 2 d after anthesis, those that got enlarged had strong expression signals; the others that were wilting had weak signals. This indicated that auxin-binding protein 1 gene possibly play a role in cucumber fruit development. When auxin-binding protein 1 gene of Arabidopsis was transformed into cucumber, the parthenocarpic rate of transgenic plants was 31.7%, higher than the control. This result showed that the sensibility to auxin was increased in transgenic plants. PMID- 15789771 TI - [Cysteamine attenuates suppression of peripheral IL-2 and lymphocytic proliferation induced by ruminal and duodenal operations in goats]. AB - Sixteen castrated male goats were randomly allocated into two groups (Control n=8; Experiment n=8) to investigate the effects of cysteamine on surgery-induced cellular immune suppression. The experiment commenced with 13 d of pre-operation (pre-Op), which contained 3 d of roughage-fed period (RP) and 10 d of roughage + concentrate-fed period (RCP), followed by first operation (Op-1) and then the second operation (Op-2). The goats were allowed to have 14 d of recovery between two operations. The fistulas were fixed to rumen walls in Op-1 and to duodenal walls in Op-2. Cysteamine (15 mg x kg(-1) x BW x d(-1)) was added to diet and fed to goats in Exp. group since RCP of pre-Op through whole experimental period. Blood was sampled through jugular vein from goats of both groups at pre-feeding of the last day of RP and RCP, 8 d after Op-1, 4 d and 13 d after Op-2, respectively to determine serum concentration of interlukin-2 (IL-2), cortisol, as well as PHA-stimulated lymphocytic proliferation rate (SI). In pre-operative periods there were no significant differences in concentration of IL-2 (0.82+/ 0.1 vs 1.03+/-0.22 microg l(-1)) and cortisol (20.48+/-4.52 vs 20.80+/-9.93 microg l(-1)), as well as lymphocytic SI (90.72+/-7.25 vs 129.22+/-18.59) between Control and Exp. groups. The concentrations of IL-2 (0.82+/-0.1 vs 0.73+/-0.08 vs 0.55+/-0.12 microg l(-1)) and lymphocytic SI (90.72+/-7.25 vs 49.31+/-6.4 vs 7.09+/-1.66) in Control were depressed acutely (P<0.05), but cortisol concentration elevated (20.48+/-4.52 vs 26.67+/-10.51 vs 32.33+/-3.29 microg l( 1)) in post-operative periods (8 d after Op-1 and 4 d after Op-2), compared with those in RCP. While they remained unaltered in Exp. group except a slight decrease of lymphocytic SI at 4 d after Op-2. The different responses of Control and Exp. groups to surgery stress led to a 80-90% higher of IL-2 and about 3 times greater of SI in Exp. than those in Control, which was contributed by cysteamine administration. Our data show that in operated goats the cellular immune was suppressed by surgery stress, cysteamine administration prevented decreases of IL-2 and lymphocytic SI and increase of cortisol induced by surgery stress, consequently attenuates surgery-induced suppression of cellular immune activity. PMID- 15789772 TI - [Involvement of nitric oxide in the effect of interleukin-2 on the contractility of rat ventricular cardiomyocytes]. AB - The aim of the present study was to investigate the effect of interleukin-2 (IL 2) on the contractility in cardiomyocytes and the underlying mechanisms. Ventricular myocytes were isolated from adult male Sprague-Dawley rats. Contractile responses were evaluated by use of the video tracking system. Contractile parameters in cardiomyocytes electrically stimulated at 0.2 Hz included peak velocity of cell shortening (+dL/dtmax), peak velocity of cell relengthening (-dL/dtmax), contractile amplitude (dL), and end-diastolic cell length. Calcium transients of ventricular myocytes were determined by the spectrofluorometric techniques. Dose-dependent inhibition in + dL/dtmax, dL/dtmax, dL and end-diastolic cell length were induced by IL-2 at 2-1000 U/ml. Pretreatment with the nitric oxide synthase inhibitor N(w)-nitro-L-arginine methyl ester (L-NAME, 100 micromol/L) and soluble guanylyl cyclase inhibitor 1H [1,2,4]oxadiazolo [4,3a]quinoxalin-1-one (ODQ, 10 micromol/L) attenuated IL-2 induced inhibition of contractility. Aminoguanidine, an inhibitor of inducible nitric oxide synthase, had no effect on the inhibition by IL-2. IL-2 at 200 U/ml decreased the amplitude of electrically induced [Ca2+]i transients of ventricular myocytes. Pretreatment with ODQ diminished IL-2-induced inhibition of amplitude of the calcium transient. In conclusion, the present study indicates a direct action of IL-2 on cardiomyocyte contraction, possibly through an increased NO production, activation of soluble guanylyl cyclase and inhibition in intracellular Ca2+ level. PMID- 15789774 TI - [VIOXX written off--and what next? COX-2 inhibitors and cardiovascular diseases]. PMID- 15789775 TI - [Genetic determinants of osteoporosis]. AB - Osteoporosis is a chronic progressive disease, characterized by higher bone resorption than bone mass accumulation. This leads to reduction of the bone mass, to microarchitectural deterioration of the bone tissue and to the increased risk of bone fracture. Osteoporosis affects predominately postmenopausal females, but it is also diagnosed at elderly males. Although osteoporosis is influenced by various environmental factors (mainly by dietary habits and physical activity), analysis of results obtained from studies with different designs (population-, family-, association- studies) confirmed, that risk of osteoporosis development and risk of osteoporotic fractures are significantly influenced by genetic predispositions. The genetic determination of osteoporosis is not yet completely understood. However, it is clear, that although there are mutations in some genes causing rare form of osteoporosis, osteoporosis is a polygenic disease--it is influenced by common variants (polymorphisms) of several different genes. For example genes for vitamin D receptor, estrogen receptor alpha, collagen type I alpha1, transforming growth factor beta-1 and some others are indicated to play a role in genetic determination of osteoporosis. In this review we summarize our recent knowledge about the genetic determination of osteoporosis. PMID- 15789776 TI - [A modern method for differential gene expression analysis and its significance for cancer diagnostics and therapy]. AB - Contemporary approaches to diagnostics and therapy in oncology are nowadays tightly coupled to novel findings of biomedical science. One of the main trends in molecular medicine is the development of methodologies enabling parallel monitoring of expression of large quantities of genes or proteins--so called functional genomics and proteomics. These techniques allow determination of differential gene expression, i.e. evaluation of differences in gene expression between two or more cell or tissue samples of different types (e.g. normal or cancer cells) or coming from different culture conditions. These approaches help in elucidating causes of malignant transformation and can serve as a base for development of targeted anticancer gene therapy, monitoring of patient response to treatment and prediction of further disease development. Genomic approaches have undergone rapid development in the last few years--from differential and subtractive hybridisation through differential display all the way to serial analysis of gene expression and DNA microarrays. Besides that, tissue and protein arrays and other proteomic approaches have been also used. Currently DNA microarrays covering expression of the whole human genome, having significant potential in oncological research and clinical praxis, have been used more and more frequently. Many new tumor growth and progression markers were found using such approaches. Some of these markers have been already successfully used in clinical practice (e.g. in breast cancer) for therapy optimisation and minimisation of patient discomfort. PMID- 15789777 TI - [The use of moderate hypothermia in neurosurgery]. AB - Hypothermia is currently considered as the most effective neuroprotective method. In recent years hypothermia has been more and more applied in clinical conditions. Hypothermia has been used with promising results in severe head trauma where it can evidently decrease the intracranial pressure, improve cerebral perfusion pressure and by its direct neuroprotective effect it diminishes the secondary ischemic brain damage. Hypothermia has been widely used also during complicated neurosurgical operations when cerebral vessels are manipulated (operations of cerebral aneurysms, arteriovenous malformations, scull base tumours). Hypothermia has been recently tested also in some types of stroke, mainly in subarachnoid haemorrhage and ischaemic stroke. PMID- 15789778 TI - [C-reactive protein in the pathogenesis of atherosclerosis: advantage and pitfalls of the "Mainz hypothesis"]. AB - C-reactive protein can be viewed as a basic marker of activity of the inflammatory response, which modulates the development and the progression of atherosclerosis including its life-threatening complications. At the same time, C reactive protein represents an active partaker or mediator of this same inflammatory reaction. However, at the very beginning of atherosclerotic disease, C-reactive protein exerts a clear-cut antiatherogenic activity. The two aspects of CRP's function, i.e. both the pro-inflammatory and the anti-inflammatory one, respectively, stem from CRP's extent of co-operation with the complement system. From the evolutional point of view, the anti-inflammatory activity of CRP is the primary one, in that it sets stage for the host to remove foreign particles and to accelerate wound healing. The influence of well-known atherogenic risk factors converts the originally beneficial influence of CRP into pro-inflammatory and pro atherogenic effects. This review article presents new conclusions from the "Mainz hypothesis". It shows that the primary protective action of CRP resides in its regulatory influence on the extent of activation of the complement system after the latter has been triggered by enzymatically remodeled low-density lipoproteins. In further course of atherosclerotic disease, C-reactive protein exhibits a full-blown proinflammatory activity. It can result in the progression of the primary morphologic lesions up to the development of sudden vascular events. PMID- 15789779 TI - [Ten years of molecular monitoring of chronic myeloid leukemia by quantitative RT PCR]. AB - Chronic myeloid leukemia (CML) is characterized by the presence of BCR-ABL fusion gene resulting from the reciprocal chromosome translocation t(9;22)(q34;q 11), karyotypically detected as Ph chromosome. BCR-ABL gene was proved to play the principal role in CML pathogenesis. It is a hallmark of CML used in diagnostics and monitoring of the response to the therapy. The most sensitive method of detecting BCR-ABL aberration is RT-PCR which is able to find a single in leukemic cell between 10(6) normal leukocytes. Monitoring of BCR-ABL transcript level by quantitative RT-PCR is of the high prognostic value. High or increasing BCR-ABL transcript number signalizes bad response to treatment and a bad prognosis. On the contrary RT-PCR negativity, low level, or decreasing BCR-ABL transcript number denotes good response to treatment and good prognosis. Q-RT-PCR can detect changes in disease status several weeks or even months earlier than other methods. In 1994 the Q-RT-PCR was introduced at the Institute of Hematology and Blood Transfusion in Prague and was used for early detection of relapse after transplantation. At present it is used in all patients with CML for monitoring of response to the treatment. We have confirmed that this precise, sensitive and non invasive method is of the principal importance for monitoring of disease status in CML patients. PMID- 15789780 TI - [Mechanical cardiac support--the first use in Czech Republic]. AB - BACKGROUND: Implantation of mechanical assist device is widely accepted modality of treatment of patients with refractory heart failure. In the present study we evaluated our first one-year experiences with this method for bridging patients to cardiac transplantation. METHODS AND RESULTS: Between April 2003 and May 2004, the Thoratec VAD (Thoratec, Pleasanton, CA, USA) was implanted in 6 patients ( males; age 28-61 years) as a bridge-to-transplant procedure after having received maximum inotropic support and who were at imminent risk of death. In all patients was performed VAD as biventricular device (BiVAD). During a week after placement was observed recovery of organs function in all patients. Five patients survived to heart transplantation. One patient died 21 days after BiVAD placement due to massive bleeding to the respiratory tract. In post-transplantation period 1 patient died second day from acute graft failure and other patient died 34 days after from intracranial bleeding. Three patients has been discharged from the hospital and they are surviving more than I year. CONCLUSIONS: Analysis of our first experiences with the Thoratec BiVAD implantation as bridging to heart transplantation suggests that it is well suited method with respect to long- term prognosis of this group of patients. PMID- 15789781 TI - [Pilot study of the noninvasive assessment of endothelial dysfunction by post occlusion dopplerometry velocity curves analysis in arteria brachialis]. AB - BACKGROUND: A new method of noninvasive assessment of endothelial dysfunction was used in this pilot study. Study has been set on a. brachialis dopplerometry velocity curves analysis. Index of blood stream deceleration between the first and second minute of the postocclusion periode was measured on a. brachialis after transient mechanical occlusion of the forearm--so called Deceleration Index. METHODS AND RESULTS: Eighteen patients with so far untreated hypercholesterolemia were involved in our study. Profile of velocity curves became significantly different after six months of fluvastatin therapy, Deceleration Index increased from 3 % to more than 7 % (p<0.05). That correlated with LDL-CH levels decrease, but also with the significant decrease of biochemical markers of endothelial dysfunction (trombomodulin, von Willebrand factor). CONCLUSIONS: The result is corresponding with previously published studies that have proved endothelial function improvement after statin therapy using another ultrasonographic methods. PMID- 15789782 TI - [Physical exercise and yoga in prevention and treatment of addictive diseases]. AB - Prevention of addictive diseases should be complex and systematic and it should include training of social skills, decision-making skills, family intervention, etc. Similarly, effective treatment is usually long-term, systematic and complex. Physical exercise and yoga can be useful components of comprehensive prevention and treatment programmes. On the other hand, competitive professional sport rather increases the number of risk factors for substance-related problems. Practical experience with the use of yoga in substance dependent patients and pathological gambles are mentioned. One of the advantages of yoga is the integration of physical exercise and relaxation. PMID- 15789783 TI - [A glance into history of angiology in central Europe]. AB - The question is: What is angiology? It is the branch of medical science studying blood and lymph vessels and their disorders. With the development of techniques, diagnostic and therapeutic methods and with the increasing morbidity and mortality on vascular diseases angiology became in the last decades an important specialization of medicine. PMID- 15789784 TI - [Albert Schweitzer and the present]. PMID- 15789785 TI - Ultrasound scan-guided core sampling for diagnosis versus freehand FNAC of the thyroid gland. AB - BACKGROUND AND AIM: Freehand fine needle aspiration cytology (FNAC) is an obligatory investigation of the thyroid nodule. Between 5.0-43.1% of FNAC samples are reported as being initially unsatisfactory. In our unit, thyroid freehand FNAs are performed with a small needle (21 or 23G). Non-dominant nodules as part of multinodular goitres, difficult to palpate nodules or nodules with previously unsatisfactory freehand FNACs are sampled under ultrasound scan (USS) guidance with the larger 20G cutting core sampling technique. We aimed to compare the satisfactory sampling rate and safety of the two different methods. PATIENTS AND METHODS: Cytology forms were reviewed for 262 freehand FNACs and USS-guided core samples, performed in our unit over a two-year interval (1 July 1999 to 30 June 2001). RESULTS: Ultrasound-guided core samples for cytology were unsatisfactory (AC0-1) in 19/121 (15.6%) of the cases, compared with 66/141 (46.8%) of freehand FNACs (p value = < 0.0001). Ten out of eleven patients (91%) had a satisfactory USS-guided core after an unsatisfactory freehand FNA; 7/15 patients (46.7%) had satisfactory repeat freehand FNACs following an initial unsatisfactory freehand FNAC (p value = 0.0191). There were no complications as a result of either freehand FNAC or USS-guided core sampling. CONCLUSION: USS-guided cores provided more satisfactory samples for assessment than freehand FNACs. The USS-guided technique is safe despite the use of the larger cuffing needle. The USS-guided core sampling was also a useful tool for repeat thyroid nodule sampling after an unsatisfactory freehand FNAC. PMID- 15789786 TI - Topical nonsteroidal anti-inflammatory drugs versus oil of evening primrose in the treatment of mastalgia. AB - OBJECTIVE: To compare oil of evening primrose (OEP) and topical nonsteroidal anti inflammatory (NSAIDs) with respect to safety, effectiveness, rapidity of response, cost effectiveness and acceptability in the treatment of breast pain. STUDY DESIGN: An open, non-randomised, comparative study of topical (NSAI) gel versus OEP was carried out, over a period of one year. PATIENTS AND METHODS: Fifty female patients attending the outpatient department with moderate to severe breast pain were given one of the two agents alternatively, after selection. RESULTS: Results showed that out of 25 patients treated with OEP, 64% had a clinically significant response after three months of treatment, compared with 92% with topical NSAIDs. Only one patient (4%) had side effects with OEP, while no patient had side effects with topical NSAIDs. Twenty per cent and seventy per cent showed acceptability as far as costs were concerned and mode of administration respectively, with OEP. The acceptability rate was 68% and 96% respectively, with topical NSAIDs. CONCLUSION: This study has shown topical NSAIDs to be safe, effective, rapid and acceptable mode of treatment for cyclical and non-cyclical mastalgia. PMID- 15789787 TI - Same admission colostomy closure: a prospective, randomised study in selected patient groups. AB - BACKGROUND: Despite a recent trend towards primary repair, colostomy still has an important place in the management of different colorectal pathologies. Timing of colostomy closure is a debatable issue among general surgeons. We carried out a prospective, randomised study to compare the outcome after same admission colostomy closure (SACC) and conventional delayed colostomy closure (CDCC) in patients with predominantly traumatic injury of the large bowel. METHODS: Sixty patients, the majority with traumatic colorectal pathologies, were prospectively randomised to SACC [30 patients, 23 men, mean age 27.9 +/- 9.7 (range, 18-65) years] or (CDCC) [30 patients, 24 men, mean age 28.6 +/- 10.6 (range, 18-63) years]. All colostomies were closed using an intraperitoneal closure technique. Pre-operative data and post-operative outcomes were collected and analysed. RESULTS: A total of seven patients (23.3%) with SACC developed complications compared with eight patients (26.6%) having CDCC (p = 0.83). The mean hospital stay (p < 0.01), as well as the overall cost (p < 0.001), were significantly less in the SACC group. CONCLUSIONS: Same admission colostomy closure is a safe and cost-effective technique for colostomy closure in selected groups of patients. PMID- 15789788 TI - Pancreas transplantation in Ireland. AB - BACKGROUND: We report on our experience of pancreatic transplants performed in Beaumont Hospital from 1992 to January 2003. During this period, 63 pancreatic transplants were carried out. Simultaneous pancreas and kidney transplantation is regarded as the treatment of choice for Type 1 diabetics with end stage renal failure (ESRF) by the American Diabetes Association. METHODS: A retrospective review of all the pancreas transplants between 1992 and January 2003 was carried out. All of the patients had insulin-dependent diabetes mellitus (IDDM) and, in addition, 57 of the recipients had concomitant end stage renal failure. For two of the patients, this was a second transplant, one of them having had a previous renal graft which thrombosed. The second patient underwent a simultaneous pancreas and kidney (SPK) transplant in 1998 with loss of the pancreatic graft shortly afterwards. She subsequently received a pancreas only transplant in 2002. RESULTS: The follow-up period ranges from one month to ten years and parameters used to assess graft function include Hb A1c and serum creatnine (SPK only). There have been 22 graft losses (six kidneys, nine pancreases and seven involving both the kidney and pancreas), to date. The one-year graft survival was 80.4% (pancreas) and 92.7% (kidney), with a one-year patient survival of 94.8%, which compares favourably with figures from other centres. At follow-up, the average Hb A1c at six months post-transplant was 5.7% with a serum creatnine of 121micmol/L being recorded at one month. CONCLUSION: Our experience of pancreas transplants at Beaumont Hospital has been very encouraging with graft and patient survival equivalent to other international centres. PMID- 15789789 TI - Experience with different treatment modules in hidradenitis suppuritiva: a study of 106 cases. AB - BACKGROUND: Hidradenitis suppuritiva, a disease of the apocrine sweat glands, has always been a challenging area to the plastic and reconstructive surgeon. The aetiology is idiopathic and medical treatment offers temporary relief only. Radical surgical excision, therefore, is the mainstay of active management. AIM: To present and evaluate a series of 106 patients of hidradenitis suppuritiva treated in the regional plastic surgery unit of southeast Scotland between January 1990 and December 2000. PATIENTS: Median age at onset was 36 years (range 17-70 years). The patients were predominantly females (88.78%) and heavy smokers (98.1%). Median age before active surgical intervention was six years (range 1-30 years). The patients were divided into a mild group (n=64) and a severe group (n = 42). All patients had broad-spectrum antibiotics and multiple incision and drainage procedures before referral from general practitioners, dermatologists or general surgeons. One hundred sites were managed by primary closure (mostly in the 'mild' group); 29 resurfaced with split skin grafts and 14 with local, fasciocutaneous or musculocutaneous flaps (mostly in the 'severe' group). Median postoperative follow-up period was three years (range 1-7 years). RESULTS: In the 'primary closure' series, recurrence rate requiring at least one secondary operation was 69.88%. There was no recurrence, no serious complications and no revision operations in the 'graft' and the 'flap' series. CONCLUSION: A high index of suspicion is required before contemplating primary closure in hidradenitis suppuritiva, even in the ' mild' variety. This condition should always be treated aggressively by radical excision of all hair-bearing areas and reconstructed with a graft or a flap. PMID- 15789790 TI - An analysis of urinary tract trauma in Scotland: imnpact on management and resource needs. AB - OBJECTIVE: We report the incidence, distribution, aetiology and outcome of urological trauma in a unique national database to provide an insight into its contemporary management. PATIENTS AND METHODS: The Scottish Trauma Audit Group prospectively collected data from severe trauma presenting to all major Scottish hospitals. We examined data from 24,666 trauma admissions from 1999 to 2002. Patients who sustained urological injuries were identified and studied in detail. RESULTS: 362 patients had urological injuries, comprising 1.5% of the trauma population, and an incidence of 1 per 45,000 head of adult population per year. Blunt injury (n = 285, 79%) was the main cause of urological trauma. Road traffic accidents were most frequent (197 patients, 54%), followed by assaults (76, 21%) and high falls (45, 12%). Renal injuries were the most common (n = 241, 67%), followed by injuries to the external genitalia (71, 20%), bladder (65, 18%), urethra (16, 4%) and ureter (3, 1%). Only 52 patients (14%) had isolated urological trauma. One hundred and fifty nine out of 310 (51%) urological patients with associated injuries were physiologically compromised on arrival in A&E, compared with only 4/52 (8%) patients with isolated urological trauma. All patients with isolated urological trauma survived, whereas 110/310 (35%) of those with associated injuries died. CONCLUSION: Urological injuries in Scotland mostly result from blunt trauma due to high-energy impacts. Isolated urological injuries are uncommon and all such patients survived. The majority of patients with urological trauma have multiple injuries and require a multi-disciplinary approach. Current urological services appear adequately distributed to cope with contemporary demands of urological trauma. PMID- 15789791 TI - An audit of ultrasound-assisted catheter insertion in patients receiving chemotherapy. AB - BACKGROUND AND AIMS: Hickman catheters have been shown to provide safe long-term venous access for patients with malignant diseases. In many centres, catheters are placed using fluoroscopic guidance. We hypothesised that ultrasound-assisted catheter placement by surgeons in the operating theatre would be a simple, safe and effective alternative technique with reduced infective complication rates. METHODS: Hickman catheter insertions between May 1998 and March 2002 were studied. The data were collected from the Hospital Inpatient Enquiry database and the case notes of all patients were reviewed. Percutaneous catheter placement with tunnelling was performed in the operating theatre after scanning the internal jugular vein (IJV) for position, size and patency, using a Pie 100LC Scanner (Pie Medical, Maastricht). A standard chest radiograph confirmed catheter position at the end of the procedure. RESULTS: Fifty-eight patients (30 males and 28 females) had 65 Hickman catheters inserted. The median age was 60 years (range 32-82 years). Catheter placement was achieved in all patients, 59 in the right IJV and six in the left. Ultrasound scanning demonstrated that the right IJV was thrombosed in six patients (10%), thus avoiding unnecessary attempts at cannulation. The Hickman catheters remained in situ for a combined total of 5857 days (median, 89 days, range 4-485 days). Immediate complications occurred in two patients (pneumothorax in both). One patient required a chest drain. The overall sepsis rate was 3.92 per 1000 catheter days. Systemic sepsis was slightly higher (2.21 per 1000 catheter days) than superficial sepsis (1.71 per 1000 catheter days). In all the patients who developed systemic sepsis the catheter had to be removed (n = 13). All the superficial infections were treated successfully with antibiotics (n = 10 patients). Two catheters developed thrombosis. CONCLUSION: We conclude that ultrasound-assisted percutaneous placement of Hickman catheters in the operating suite is a simple, safe and effective technique and may help to reduce infective complications. PMID- 15789792 TI - Laparaoscopic cytogastrostomy for a giant pseudocyst of pancreas. AB - BACKGROUND: Conventionally, a large symptomatic and unresolved pancreatic pseudocyst is treated surgically by internal drainage to a neighboring adherent viscus such as stomach, duodenum or jejunum. Recently, the various minimal invasive approaches have been used to treat this condition. Depending on the expertise available, the cyst can be also be drained endoscopically or laparoscopically. We present a case of a large pseudocyst treated laparoscopically. METHOD: A 60-year-old lady was admitted for an elective laparoscopic cholecystectomy as a day case. Under general anaesthesia during her elective laparoscopic cholecystectomy a mass was visible and palpable in the left upper abdomen. Post-operatively, a CT scan of abdomen confirmed the presence of a giant pseudocyst of the pancreas. She successfully underwent a laparoscopic cystogastrostomy four weeks later. CONCLUSION: Laparoscopic cystogastrostomy for pseudocyst of the pancreas is safe, feasible and with good outcome. PMID- 15789793 TI - An unusual case of traumatic petechiae. AB - We report a rare cause of traumatic petechiae without the full blown symptoms and signs of traumatic asphyxia. There were no complications encountered and there was spontaneous resolution. PMID- 15789794 TI - Jaundice as a presentation of phenol induced hepatotoxocity following injection sclerotherapy for haemorrhoids. AB - A 43-year-old man was admitted with jaundice six days following phenol injection sclerotherapy for haemorrhoids. He was diagnosed with a phenol-induced hepatitis. Although he remained well, liver function tests only returned to normal after six months. Systemic absorption of phenol has been reported with ingestion, upper airway and excessive cutaneous exposure but not as a complication of haemorrhoidal injection sclerotherapy. Hepatic involvement is also rare and usually the result of ongoing sepsis. We report the unique case of a patient presenting with jaundice secondary to chemical hepatitis, following systemic absorption of phenol at injection sclerotherapy. This case highlights the importance of clinical awareness of not only the infective complications of injection sclerotherapy but also the potential for phenol to be absorbed systemically with severe consequences. A brief overview of symptoms of phenol toxicity is included. PMID- 15789795 TI - Bilateral ureteral obstruction after appendicectomy. AB - Acute renal failure due to bilateral ureteral obstruction is a rare complication after appendectomy in children. We report a case of anuria in an 11-year-old boy five days after surgery for a perforated acutely inflamed appendix. No abscess was found with CT. After saline-filling of the urinary bladder, transabdominal ultrasound demonstrated echogenic "plugs" in the distal parts of both ureters. Cystoscopy revealed inflammatory changes in the bladder base. Following introduction of bilateral ureteric stents, there was rapid normalisation of urinary output and serum creatinine. A review of the literature disclosed 15 similar cases, all boys in the age group 6 to 15 years. PMID- 15789796 TI - Ovulating ovary in an inguinal hemia. AB - BACKGROUND: An ovary can, occasionally, be found in an inguinal hernia in the newborn female. This is extremely rare in premenopausal women. We report a case of an ovulating ovary in an incarcerated inguinal hernia. There is only one similar case of incarcerated ovulating ovary reported in the literature. METHOD AND FINDINGS: A 23-year-old Turkish woman, mother of three children, presented with a painful swelling over the upper part of left labia majora of 48 hours duration. She was diagnosed as having an obstructed left inguinal hernia. At operation the left ovary and fallopian tube were seen in the left inguinal canal. The ovary and fallopian tube were congested but viable and mature ovarian follicles could be seen on the ovary. A biopsy was taken from the ovary, the ovary and the fallopian tube were easily reduced back into the abdominal cavity and the hernia repaired. DISCUSSION: The presence of an ovary in the inguinal canal is very rare and should be a differential diagnosis in women presenting with an irreducible lump in the inguinal area. It should be treated as a surgical emergency. PMID- 15789797 TI - Spontaneous apical lung herniation presenting as a neck lump in a patient with Ehlers-Danlos syndrome. AB - Ehlers-Danlos syndrome (EDS) is a heritable group of disorders of connective tissue characterised by skin hyperlaxity, joint hypermobility and tissue fragility. We report on a 49-year-old man with EDS who presented with spontaneous swelling at the root of the right neck during coughing. The size and site of the lesion led to diagnostic uncertainty. Subsequent computerised tomography (CT) scanning during a Valsalva manoeuvre demonstrated a large apical lung hernia arising through the costo-vertebral fascia into the root of the neck. No previous reports have documented spontaneous apical lung herniation in patients with EDS. This case serves to remind all practitioners dealing with the head and neck to remember the apical lung as a potential source of pathology in the neck. PMID- 15789798 TI - Stump appendicitis: a rare but important entity. AB - A case of right lower quadrant pain in a 35-year-old male who underwent an appendicectomy 14 years previously is presented. Recurrent appendicitis with perforation was noted in an appendiceal stump on exploratory laparotomy. Although rare, inlammation of the appendiceal stump can occur and is still an important clinical entity. It is difficult to diagnose pre-operatively. A wide spectrum of causes in the differential diagnosis of right lower quadrant pain of the abdomen and a previous appendicectomy in a patient's history may delay the diagnosis. Knowledge of the condition should permit the physician to make an early diagnosis and, thus, limit the resultant morbidity. PMID- 15789799 TI - Re: Primary total hip replacement Surg J R Coll Surg Edinb Irel, 1: 2003; 332 341. PMID- 15789800 TI - Re: Anatomy: a must for teaching the next generation. Surg J R Coll Surg Edinb Irel 2004; 2(2): 79-90. PMID- 15789801 TI - Re: Registrar operating experience over a 15-year period: more, less or more or less the same? Surg JR Coll Edbinb, Irel 2004; (2)3: 161-64. PMID- 15789802 TI - Re: Efficacy and safety of peri-prostatic local anaesthetic injection in transrectal biopsy of the prostate: a prospective, randomised study. PMID- 15789803 TI - Re: Bannister GC. letters. Surg JR Coll Edinb Irel 2004; 2 (4): 243. PMID- 15789804 TI - Re: Luscombe KL, N Maffulli. The three-in-one procedure: how I do it. Surg J R Coll Edinb Irel 2004; 1: 32-36. PMID- 15789805 TI - Evolution and revolution. PMID- 15789806 TI - Use of a multi-stage exercise test to assess the responsiveness of rate-adaptive pacemakers in dogs. AB - OBJECTIVES: To assess the ventricular rate response of rate-adaptive (VVIR) pacemakers in dogs using a multi-stage exercise test. METHODS: The rate responsiveness of VVIR pacemakers was assessed in seven dogs with complete atrioventricular (AV) block and implanted with various models of pulse generators (six motion sensors and one automatic dual-sensor rate-response pacemaker). Response activity was assessed with a multi-stage exercise test on a treadmill. Atrial and ventricular rate were analysed retrospectively at the end of the test and the AV ratio was calculated after each minute of exercise. RESULTS: During exercise, the mean (sd) AV ratio recorded in all paced dogs was 1.7 (0.5) (expected physiological ratio 1.0), although a variety of individual performances was observed. A poor response (AV ratio 2.8 [0.2]) was obtained with the automatic dual-sensor pacemaker, suggesting that this type of rate-responsive device may not be indicated for implantation in dogs with complete AV block. The overall AV ratio for the six dogs implanted with motion sensors was 1.4 (0.2), showing a better performance of these pacemakers during exercise. CLINICAL SIGNIFICANCE: This multi-stage exercise test represents an easy and repeatable method for assessing the accuracy of rate-responsive sensors and offers valuable information for the correct setting of VVIR pacemakers in dogs. PMID- 15789807 TI - Efficacy and safet of pimobendan in canine heart failure caused by myxomatous mitral valve disease. AB - OBJECTIVES: To evaluate the clinical efficacy and safety of pimobendan by comparing it with ramipril over a six-month period in dogs with mild to moderate heart failure (HF) caused by myxomatous mitral valve disease (MMVD). METHODS: This was a prospective randomised, single-blind, parallel-group trial. Client owned dogs (n = 43) with mild to moderate HF caused by MMVD were randomly assigned to one of two groups, which received either pimobendan (P dogs) or ramipril (R dogs) for six months. The outcome measures studied were: adverse HF outcome, defined as failure to complete the trial as a direct consequence of HF; maximum furosemide dose (mg/kg/day) administered during the study period; and any requirement for additional visits to the clinic as a direct consequence of HF. RESULTS: Treatment with pimobendan was well tolerated compared with treatment with ramipril. P dogs were 25 per cent as likely as R dogs to have an adverse HF outcome (odds ratio 4.09, 95 per cent confidence interval 1.03 to 16.3, P = 0.046). CLINICAL SIGNIFICANCE: R dogs had a higher overall score and thus may have had more advanced disease than P dogs at baseline (P = 0.04). These results should be interpreted cautiously but such a high odds ratio warrants further investigation. PMID- 15789808 TI - Incidence of post-anaesthetic arrhythmias in dogs. AB - OBJECTIVES: To assess the use of Holter monitoring for evaluating the incidence of post-anaesthetic cardiac arrhythmias and associated anaesthetic risk for two different anaesthetic protocols. METHODS: Patients undergoing orthopaedic surgery were randomly divided into two groups with different anaesthetic regimens (group A, isoflurane n = 30; group B, propofol n = 30). Two 24-hour Holter recordings were performed for each patient: the first directly following anaesthesia and the second, as a comparison, on the fifth postoperative day. RESULTS: Although all dogs were healthy on pre-anaesthetic cardiac evaluation, 56 dogs showed arrhythmias in the two 24-hour (Holter) electrocardiograms performed. However, the number of arrhythmias recorded was low in most cases (less than 10 supraventricular extrasystoles and less than 100 ventricular extrasystoles). One patient in group A showed 94 supraventricular extrasystoles during the second monitoring period. Three patients in each group developed more than 100 ventricular extrasystoles during both Holter recordings. There were no statistically significant differences between the two anaesthetic regimens or between the two recordings in both groups. CLINICAL SIGNIFICANCE: The two anaesthetic protocols investigated in this study did not induce an increased incidence of severe arrhythmias in healthy dogs in the post-anaesthetic phase. PMID- 15789809 TI - Cardiac troponin I: evaluation I of a biomarker for the diagnosis of heart disease in the dog. AB - OBJECTIVES: To assess the value of measuring blood levels of the myocardial protein cardiac troponin I (cTnl) in the diagnosis of congenital and acquired heart disease in the dog and in the evaluation of the severity of heart failure. METHODS: Serum samples obtained from healthy dogs (n = 26) and from dogs diagnosed with a variety of congenital and acquired heart conditions (n = 35) were assayed for cTnl concentration using an automated immunoassay method. Results were also analysed according to the degree of heart failure as assessed using the International Small Animal Cardiac Health Council's scheme. RESULTS: Healthy dogs had very low or undetectable blood cTnl levels, as did dogs with congenital heart disease. However, cTnl levels were significantly elevated in dogs with acquired mitral valve disease, dilated cardiomyopathy and pericardial effusion. Blood cTnl levels also varied with severity of heart failure. CLINICAL SIGNIFICANCE: Measurement of blood cTnl levels may be a useful aid in the diagnosis of dogs with suspected heart disease and in indicating the severity of heart failure. PMID- 15789810 TI - Compound maxillary odontoma in a young German shepherd dog. AB - A three-month-old, male German shepherd dog was admitted with a facial mass of two months' duration. Clinical examination showed a round mass, 3 cm in diameter, in the left infraorbital area. The upper last premolar deciduous tooth was not erupted. No other abnormalities were detected. Radiological examination revealed a posterior maxillary mass of mixed opacity. The mass was surgically excised. Histopathological examination demonstrated a connective tissue stroma containing foci of irregular enamel and dentine, resembling rudimentary teeth (denticles), surrounded by new bone formation. Morphology and structure of the denticles were also confirmed by scanning electron microscopy. A compound odontoma was diagnosed. One year after surgery, the dog was free of clinical signs. PMID- 15789813 TI - Responding to the draft veterinary medicines regulations. PMID- 15789812 TI - What is your diagnosis? Tricuspid valve stenosis (TS). PMID- 15789811 TI - Arrhythmogenic right ventricular cardiomyopathy in two cats. AB - Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a disease characterised by infiltration of the myocardium by adipose and fibrous tissue. The disease is an important cause of sudden death in humans, but has rarely been described in animals. This report describes ARVC in two cats with right-sided congestive heart failure. One cat had also experienced previous episodes of syncope. Standard six lead and 24-hour (Holter) electrocardiogram recording revealed complete atrioventricular block and multiform ventricular ectopics in both cats, with the addition of ventricular tachycardia, ventricular bigeminy and R-on-T phenomenon in one of them. On echocardiography, the right ventricle and atrium were massively dilated and hypokinetic. The survival times of the cats were three days and 16 days following diagnosis. Histopathology in one case revealed fibro-fatty infiltration of the myocardium, predominantly affecting the right ventricular free wall. PMID- 15789814 TI - Saving a life in a crisis. PMID- 15789815 TI - Spreading excellence. PMID- 15789817 TI - [Methodology and quality control in EPIGRAM, a pharmacoepidemiological study of obesity in general practice]. AB - The EPIGRAM pharmacoepidemiological study was conducted in general practitioners prescribing orlistat (Xenical) in order to describe, under real clinical conditions, the management of obese or overweight patients. A 1-year follow-up of a patient cohort treated with orlistat was also conducted. Quality criteria specific to the study were defined to ensure the scientific validity of the results; these criteria mainly involved the following items: independent scientific committee, sampling investigator's recruitment, study organisation and quality control of the collected data. On-site control visits of a sample of 13% of participating GPs revealed that 75% of the controlled data were documented in a source file of which 85% were consistent with the source data. Only 1% of the 130 000 controlled data (data were controlled only on visits for which information was available) gave rise to correction requests. Of the patients treated with orlistat on inclusion, 17% were lost to follow-up by their GP and 25% were contacted directly at the end of the study. PMID- 15789819 TI - [Childhood poisoning: data from the French Poison Control and Toxicovigilance Centres (2002)]. AB - MATERIALS AND METHODS: Data for 2002 from a number of French poison control centres were analysed in terms of several age groups; 87 678 possible or established poisoning cases were counted, including 49 355 cases in the 0- to 18 year age group. RESULTS: The 0- to 3-year age group accounted for 71.7% of cases of childhood poisoning. Accidental poisoning was predominant up to 12 years, while from 13 to 18 years poisoning was essentially deliberate. The principal route of exposure was oral and involved liquids for babies and solid products for older children. The toxic agents most often implicated were pharmaceuticals and domestic products. The place of poisoning was mainly the home. A 'no-risk' evaluation was performed, and varied between 41.6% before the age of 3 years to 18.0% for the 13- to 18-year age group. The oldest children were more often managed in medical facilities. DISCUSSION: Fortunately, most of these poisoning cases were not serious (death rate: 0.026%). The poison control centres' information system is continuing to expand: it will allow an improvement in medical health monitoring associated with poisoning. PMID- 15789818 TI - [Use of the health insurance database by the centres for evaluation and information on pharmacodependance: examples, interests and future prospects]. AB - In France, the observation and evaluation of drug abuse and dependence associated with pychoactive medications are the responsibility of the National Commission for Narcotics and Psychotropic Drugs. In order to assist this commission, several centres for evaluation and information on pharmacodependence (CEIP) were created throughout in France. Recently, in order to complete their epidemiological tools, several centres have developed another pharmacoepidemiological approach using data for refunded prescriptions obtained from the local and regional French Health Insurance database. This article underlines the potential contribution of the Health Insurance database to improving knowledge of drug use in the real-life conditions based on studies performed by the CEIP. Several examples are given showing the extent of the possibilities (population-based studies, cohort studies, development of misuse indicators). In spite of their limitations (e.g. the difference between consumption and delivery), these examples confirm that these database may be a novel tool for CEIP to assess a potential abuse of a medication. PMID- 15789820 TI - [Abuse of licit and illicit psychoactive substances in children and teenagers in the PACA Region (Southeastern France)]. AB - INTRODUCTION: The abuse of licit and illicit psychoactive substances in children and teenagers represents a public health problem. MATERIALS AND METHODS: The aim of this study was to evaluate the extent of this phenomenon and its growth in the PACA (Provence-Alpes-Cote d'Azur) region (Southeastern France). The cases identified during the 10-year period 1992-2002 by the CEIP (Centre d'Evaluation et d'Information sur la Pharmacodependance) were studied retrospectively. RESULTS: One hundred and ninety-one cases were identified. In most instances, abuse involved a first experience or was sporadic. The users were often boys (70%). The substances were solvents (48.7%) used collectively (33%) during recreational activities (74%). Illicit drugs accounted for 28.2% of the cases involving a first experience (89%), with polyconsumption accounting for 50%, cannabis 57.6%, ecstasy 25.9%, and LSD (lysergic acid diethylamide) 12.1% of cases. Medicines were also used (16%), the most frequent being benzodiazepines (51.6%). CONCLUSION: The study confirms the abuse of licit and illicit psychoactive substances in children and teenagers and provides information regarding the substances available in this region of France. PMID- 15789821 TI - [Pharmaceutical development aimed at improving safety for children]. AB - The safety of paediatric drugs should be improved so that the number of child deaths due to the misuse of different kinds of medication can be decreased. Formulations, excipients and containers are the major areas that should be optimised in the pharmaceutical development of a paediatric drug. Today, Afssaps (Agence francaise de securite sanitaire des produits de sante) and the European regulations play a major role in promoting paediatric medications. PMID- 15789822 TI - Child overdose: the value of the French pharmacovigilance causality assessment method. AB - BACKGROUND: In France, the regional pharmacovigilance centres manage drug overdose as adverse drug reactions (ADRs) using the French ADR causality assessment method, and some poison control centres (e.g. in Paris) do likewise for the most serious cases. AIM: The aim of the study was to analyse and compare the chronological and semiological scores calculated with this method, in cases of drug overdose and ADRs in children recorded in the French pharmacovigilance ADR database. RESULTS: In total, 7963 cases were analysed. The distribution of semiological criteria in drug overdose and ADR cases differed significantly (G test), with a similar result for chronological criteria (but to a lesser degree). CONCLUSION: The distinction of two types of criteria in the French ADR causality assessment method appears useful for analysing cases of drug overdose in children. This finding triggers a renewed interest in the use of this method of causality assessment of drug overdose in children, and possibly in adults. PMID- 15789823 TI - Value of in vitro models for the assessment of drug-induced haematotoxicity. AB - BACKGROUND: A new antipsychotic compound induced unexpected red cell hypoplasia (reticulocytopenia, red marrow hypoplasia) in rats dosed orally for 7 days. MATERIALS AND METHODS: Since an erythropoietin-mediated pathogenesis was excluded, in vitro tests on rat and human bone marrow cells were performed with measurement of formation of late erythroid (CFU-E) and granulocyte-macrophage (CFU-GM) colony-forming units after incubation with the drug. CFU-E together with growth factors were cultured for 2 days (rat) or 7 days (human) and CFU-GM was cultured for 7 days (rat) or 10 days (human). RESULTS: The drug induced inhibition of erythroid progenitors and myeloid progenitors for both species from 3 x 10(-5) mol/L, with the concentration inhibiting the growth of 50% (IC50) consistent with drug plasma levels measured in rats. CONCLUSION: These cloning assays on rat bone-marrow cells were shown to be adequate models for determining the haematotoxicity of the agent and to be predictive of human toxicity. With only a small amount of compound required, they can be used as screening tools to detect haematotoxic potential of candidate drugs. PMID- 15789824 TI - [Adverse drug reaction reporting in emergency medicine]. AB - METHODS: A regional survey was performed between June and September 2002, to evaluate knowledge and attitudes of emergency physicians regarding adverse drug reaction (ADR) reporting in a French district. 100 questionnaires completed by physicians working in emergency departments and/or mobile intensive care units were analysed. RESULTS: The frequency of ADRs encountered by emergency practitioners was estimated at > or = 0.73 per year and per physician. The ADR notification rate in emergency medicine was estimated at < or = 6%. A minority of physicians were responsible for the majority of ADR reporting. Sixty-four percent of emergency physicians underestimated the conditions required for ADR notification: 28% thought that certain causality was an absolute necessary condition for notification, while 37% considered that notification was required only for ADRs that were both severe and unexpected. CONCLUSION: Interventions focused on advertising ADR reporting procedures could help to improve the notification rate in emergency medicine. PMID- 15789825 TI - [Taking drugs in the working environment: a study in a sample of 2106 workers in the Toulouse metropolitan area]. AB - PURPOSE: The aim of this study was to investigate the frequency and characteristics of "supportive" drug use and addictive behaviour in the working environment. METHODS: In May 2000, 2106 workers were asked to complete an anonymous self-questionnaire during their annual compulsory examination to assess their aptitude for work. Data collected concerned socioprofessional characteristics, perceived stress, job satisfaction and satisfaction outside of the workplace, and the use of licit psychoactive substances (alcohol, coffee, drugs) to face up to job strain. RESULTS: One-third of workers used drugs in the context of work: 20% used drugs to be in good form at work, 12% used drugs at the workplace for an awkward symptom, and 18% used drugs to relax after a difficult day's work. This use concerned mainly psycholeptic drugs, which were combined with caffeine and alcohol. This behaviour and nicotine dependence were more frequent in employees and manual workers. Apart from alcohol use, psychological distress and female gender were associated with such behaviour. CONCLUSIONS: The use of medications or other licit psychoactive substances in order to face up to work difficulties is a frequent phenomenom, which concerned one-third of this sample of the French working population. Our results give an initial estimation of this little known "doping" behaviour. PMID- 15789826 TI - [Chiral switch: towards a better benefit-risk ratio?]. AB - The chirality of drugs has emerged as a major theme in drug design, discovery, development and intellectual property. Single enantiomers offer some advantages and two options are available: the switch from an existing racemic drug to the single enantiomer or the development of an enantiomerically pure chiral drug. At present, the second possibility is systematically considered. The possibility of chiral switch has led to innovative drugs. However, this opportunity may be limited by several factors: the spontaneous interconversion of the two enantiomers, the pharmacokinetic profile or the selective toxicity of the most active enantiomer, and the difficulty of demonstrating the clinical superiority of one enantiomer over the racemate. Finally, the small size of the market, the policies of the drug agencies and/or the patent problem also limit the development of enantiomers. All these factors have led the pharmaceutical industry to develop new galenic forms rather than enantiomers when the patent of the racemate is expiring. PMID- 15789827 TI - [A survey on the position of France in international clinical research as assessed by pharmaceutical laboratories]. AB - In order to evaluate the attractiveness of France for conducting international clinical trials, a survey was performed among pharmaceutical companies that are based in France or that have affiliates in France. The survey concerned international phase II and III clinical studies carried out in 2002 and 2003. Ten pharmaceutical companies representing 36% of the French market completed the survey. 134 trials were analysed in total. France recruited 8.3% of the overall number of patients recruited, and 15.0% of those recruited within Europe. France was within the overall mean with regard to the percentage of active centres (78.5% versus 79.5%) and the percentage of patients evaluable according to protocol (86.8% versus 87.3%). In contrast, France ranked within the last third of analysed countries with respect to the speed of recruitment (1.5 versus 1.9 patients/centre/month), and the number of queries per observation (16.8 versus 10.9). The analysis of the qualitative indicators of performance showed that, although the perception of pharmaceutical companies towards the quality of French medicine and administrative authorities is positive, France notably needs to improve the productivity of its clinical research in order to enhance its attractiveness for the pharmaceutical sponsors of clinical trials. PMID- 15789828 TI - Photo-irradiated curcumin supplementation in streptozotocin-induced diabetic rats: effect on lipid peroxidation. AB - BACKGROUND: Diabetes mellitus is one of the most common endocrine disorders. A large number of studies are in progress to identify natural substances that are effective in reducing the severity of diabetes. Although a number of drugs are currently marketed, their long-term use can cause a number of adverse effects. MATERIALS AND METHODS: In the present study, we examined the effect of photo irradiated curcumin on experimental diabetes in order to evaluate the antihyperglycaemic effects of this compound on streptozotocin (40 mg/kg bodyweight)-induced diabetes. Photo-irradiated curcumin was given at a dose of 10, 30 and 80 mg/kg bodyweight. The level of blood glucose was elevated in the diabetic animals. The liver, kidney and brain were assayed for the degree of lipid peroxidation, reduced glutathione content and the activity of enzymic and levels of non-enzymic antioxidants. RESULTS: Antioxidant status decreased in the diabetic animals. Oral administration of photo-irradiated curcumin for 45 days resulted in a significant decrease in the levels of blood glucose, together with near normalisation of enzymic activity and the markers of lipid peroxidation. The best results were obtained in rats treated with 30 mg/kg bodyweight of photo irradiated curcumin. PMID- 15789829 TI - [Patient influence in drug imputability: a report of 17 cases]. PMID- 15789830 TI - [Aspirin (acetylsalicylic acid)-induced thrombocytopenia, an unknown adverse effect]. PMID- 15789831 TI - [Allergic respiratory disorders and vein sclerosis with lauromacrogol 400]. PMID- 15789832 TI - [Ofloxacin-induced achilles tendinitis in the absence of a predisposition]. PMID- 15789833 TI - [Monitoring of iatrogenic neutropenia: importance of the pharmacovigilance analysis]. PMID- 15789834 TI - [Cholestasis soon after administration of amoxicillin-clavulanic acid]. PMID- 15789835 TI - [Improved intervention against vitamin D deficiency and osteoporosis required]. PMID- 15789836 TI - [Can "food pyramids" solve the obesity epidemics?]. PMID- 15789837 TI - [Calcium homeostasis and normal bone remodeling]. PMID- 15789838 TI - [The cytokine system of bone tissue]. PMID- 15789839 TI - [Normal ranges and quality assurance for dual energy X-ray absorptiometry scanning]. PMID- 15789840 TI - [Treatment of osteoporosis with antiresorptive drugs]. PMID- 15789841 TI - [Non-skeletal effects of vitamin D deficiency]. PMID- 15789842 TI - [Treatment of pain caused by osteoporotic vertebral compression fractures]. PMID- 15789843 TI - [The vitamin D status of the Danish population needs to be improved]. PMID- 15789844 TI - [Glucocorticoid-induced osteoporosis. Occurrence, prophylaxis and treatment]. PMID- 15789846 TI - [Investigation and diagnosis of familial hypocalciuric hypercalcemia in Denmark]. PMID- 15789845 TI - [Osteoporosis in men--diagnosis and treatment]. PMID- 15789847 TI - [Primary hyperparathyroidism--occurrence, symptoms, complications and treatment]. PMID- 15789848 TI - [Diagnostic imaging in primary hyperparathyroidism]. PMID- 15789849 TI - [Ultrasound-guided percutaneous parathyroid ablation]. PMID- 15789850 TI - [New techniques in the treatment of primary hyperparathyroidism]. PMID- 15789851 TI - [Parathyroid hormone-related peptide induced humoral hypercalcemia of malignancy]. PMID- 15789853 TI - [The new specialist education]. PMID- 15789852 TI - [A scientific evaluation of the "alternative" food pyramid]. PMID- 15789854 TI - Progress in the control of yellow fever in Africa. PMID- 15789855 TI - Rapid health response, assessment and surveillance after a tsunami, Thailand, 2004-2005. PMID- 15789857 TI - Diagnosis, course, and assessment of alcohol abuse and dependence in adolescents. AB - Risk for the onset of an alcohol use disorder (AUD) peaks during adolescence and the transition to young adulthood, highlighting the public health significance of alcohol use by adolescents. This chapter summarizes recent research on the diagnosis, course, and assessment of adolescent AUDs. This review focuses on developmental considerations in assessment of AUD criteria, the prevalence of DSM IV AUDs among adolescents, typical alcohol symptom profiles in youth, and limitations of DSM-IV AUD criteria when applied to adolescents. In addition, studies of AUD course in adolescents, as well as factors influencing the course of AUDs are summarized. The chapter also provides an overview of brief alcohol screening instruments and other measures used in more comprehensive assessment of AUDs in adolescents. PMID- 15789858 TI - Initiation and course of alcohol consumption among adolescents and young adults. AB - This chapter takes a normative developmental perspective on the etiology of alcohol use, focusing on the initiation and course of alcohol use (rather than alcohol use disorders) during adolescence and early adulthood. We review evidence regarding the sequelae and meaning of the age of initiation of alcohol use, consider variable- and pattern-centered approaches to modeling trajectories describing the course of alcohol use across adolescence and young adulthood, and offer developmental conceptualizations of risk and protective factors for alcohol use and related problems. PMID- 15789859 TI - High risk adolescent and young adult populations: consumption and consequences. PMID- 15789860 TI - Alcohol consumption and its consequences among adolescents and young adults. PMID- 15789861 TI - Drinking among college students. Consumption and consequences. PMID- 15789863 TI - Adolescent alcohol drinking and its long-range consequences. Studies with animal models. AB - This chapter reviews findings, mainly obtained from the selectively bred alcohol preferring (P) line of rats, on (a) the development of alcohol drinking during the peri-adolescent period, (b) neurobiological factors that may contribute to adolescent drinking, (c) interventions to prevent alcohol drinking during adolescence, and (d) some long-lasting consequences of adolescent alcohol drinking. The findings indicate that P rats readily initiate alcohol drinking during the early post-weaning, adolescent and peri-adolescent periods of development. The early age-of-onset of alcohol drinking in the P compared to the NP line is associated with (a) higher densities of serotonin-1A (5-HT1A) receptors in cerebral cortical and hippocampal regions; (b) lower densities of dopamine (DA) D2 receptors in the ventral tegmental area (VTA); (c) higher functional activity in several limbic, cortical and hippocampal regions; and (d) sensitivity to the low-dose stimulating effect of ethanol. Conditioned taste aversion (CTA) training during adolescence produces long-term effects on preventing high alcohol drinking behavior of P rats. Alcohol drinking during peri adolescence by P rats produces long-lasting effects that increase the acquisition of ethanol self-administration in adulthood, and, in addition, increase craving like behavior and the potential for alcohol relapse. With suitable animal models, a better understanding of the mechanisms underlying adolescent alcohol drinking and its long-range consequences can be attained. PMID- 15789864 TI - Adolescence. Alcohol sensitivity, tolerance, and intake. AB - Research conducted in laboratory animals has shown adolescents to be less sensitive to numerous ethanol effects that may serve as cues to limit intake, including effects evident during intoxication (e.g., ethanol-induced motor impairment, anxiolysis, social impairment, and sedation), as well as during the post-intoxication period (e.g., "hangover"-associated anxiogenesis). Conversely, adolescents are more sensitive than adults to a few ethanol effects, including ethanol-induced social facilitation and impairments in hippocampal long-term potentiation. These age-specific ethanol sensitivities are not simply related to developmental differences in ethanol pharmacokinetics. Instead, they appear related in part to an ontogenetic decline in expression of within session (acute) tolerance and to differential rates of development of neural systems underlying different actions of ethanol. Relatively high levels of ethanol intake often seen in adolescent rodents and their human counterparts may be related not only to an attenuated sensitivity of adolescents to negative cues that normally serve to limit drinking, but also their greater sensitivity to both the facilitation of social behavior by ethanol and the stimulation of ethanol intake by social experiences. Although data are sparse, studies in laboratory animals hint that under some circumstances chronic adolescent exposure to ethanol may influence ongoing neural maturation and later neural, cognitive, and behavioral functioning, including later sensitivity to and propensity to use ethanol. Recommendations for further research are discussed. PMID- 15789865 TI - Age-related effects of alcohol on memory and memory-related brain function in adolescents and adults. AB - As detailed in this brief review, there is now clear evidence that adolescence represents a unique stage of brain development. Changes in brain organization and function during adolescence are widespread, and include intense rewiring in the frontal lobes and other neorcortical regions, as well as changes in a litany of subcortical structures. Recent research suggests that, because of these changes in brain function, drugs like alcohol affect adolescents and adults differently. The available evidence, much of it from research with animal models, suggests that adolescents might be more sensitive than adults to the memory impairing effects of alcohol, as well as the impact of alcohol on the brain function that underlies memory formation. For instance, when treated with alcohol, adolescent rats perform worse than adults in spatial learning tasks that are known to require the functioning of the hippocampus. Alcohol disrupts hippocampal function, and does so more potently in adolescents than adults. In contrast, adolescents appear to be far less sensitive than adults to both the sedative and motor impairing effects of alcohol. While research on this topic is still in its infancy, the findings clearly suggest that adolescence represents a unique stage of sensitivity to the impact of alcohol on behavior and brain function. PMID- 15789866 TI - The human adolescent brain and alcohol use disorders. PMID- 15789868 TI - Comprehensive approaches to prevent adolescent drinking and related problems. PMID- 15789869 TI - Prevention of adolescent alcohol problems in special populations. AB - Research on the prevention of alcohol abuse in America has only recently begun to consider the needs of special populations. This chapter will consider alcohol prevention as a function of four major special population divisions: gender, ethnicity, region (population density), and socioeconomic status. Specific ethnic groups examined will include Hispanics, African-Americans, Asian-Americans, and Native Americans. In general, there is some support for the utility of current alcohol prevention approaches on special populations. Much population-specific work completed to date has not been rigorously designed or evaluated, though it appears likely that partnering with population gatekeepers and showing cultural respect and sensitivity to the population, and providing material that is relevant to the population's adaptation to their environment are essential. PMID- 15789870 TI - Prevention of college student drinking problems. A brief summary of strategies and degree of empirical support for them. PMID- 15789871 TI - Policies to reduce underage drinking. A review of the recent literature. PMID- 15789872 TI - Prevention for children of alcoholics and other high risk groups. PMID- 15789874 TI - Treatment of adolescent alcohol-related problems. A translational perspective. AB - In the present chapter we have sought to articulate a translational perspective linking development during adolescence to alcohol treatment and its evaluation. Clearly adolescents with alcohol use disorders cannot be treated as younger versions of adults. The unique demands of their developmental stage permeate all aspects of their behavior, including alcohol use and abuse. Multi-dimensional developmental models help inform the design, implementation, and evaluation of alcohol and drug treatment programs. While an understanding of individual risk factors is informative, only through models integrating multiple risk pathways embedded in the context of adolescent development can we hope to build more efficacious and effective systems of intervention for youth with AUDs. Effective treatment of adolescent AUDs not only urges translation of the developmental perspective into pre-existing treatment approaches but argues for a paradigm shift towards alternative intervention designs and evaluation procedures and foci. PMID- 15789875 TI - Treatment of co-occurring alcohol, drug, and psychiatric disorders. AB - Comorbid psychiatric disorders and drug use disorders (DUDs) are common among adolescents with alcohol use disorders (AUDs). These comorbid disorders have a large potential significance on the clinical course of the AUDs among adolescents, and can predict a shorter time to relapse of alcohol use. The use of medication for treatment of the various comorbid adolescent populations has increased dramatically in recent years, despite the lack of double-blind, placebo controlled studies that demonstrate their safety and efficacy. Consequently, to date, no empirically proven treatment exists for most of these comorbid disorders. This chapter reviews the state of the art regarding the treatment of comorbid adolescents. This chapter also identifies gaps in knowledge regarding the treatment of comorbid adolescents, and outlines directions for future research in this field. PMID- 15789876 TI - A brief history and some current dimensions of adolescent treatment in the United States. AB - Resources for the treatment of adolescent substance use disorders have increased over the past century in tandem with the increased visibility and cultural alarm regarding adolescent substance-related problems. The United States now has a multi-branched and growing system of adolescent treatment services that spans public and private sectors and offers services in both specialty and non specialty service settings. Most adolescents are entering treatment due to alcohol and/or cannabis-related problems (and, to a lesser degree, other illicit drugs), but present with a wide array of co-occurring problems and obstacles to recovery. Multiple levels of specialized care are available but most adolescents being treated via outpatient counseling. The number and methodological rigor of adolescent treatment outcome studies have increased dramatically in recent years. The findings of these studies suggest the need for earlier systems of problem identification and intervention, a model of sustained recovery support for adolescents presenting with high problem severity and complexity, and sustained interventions with the adolescent's post-treatment family and social environment. In the opening decade of the twenty-first century, the treatment of adolescent substance use disorders is itself maturing into a professionalized and science guided service arena. PMID- 15789877 TI - Evidence-based cognitive-behavioral and family therapies for adolescent alcohol and other substance use disorders. PMID- 15789878 TI - Assessment issues in adolescent drug abuse treatment research. AB - Experimentation with alcohol and other drugs (AOD) is commonplace among American adolescents. Despite reduction efforts, the use of AOD by adolescents has increased over the past decade. A number of youth experience significant negative personal, societal, economic, and health ramifications, but continue to abuse AOD and develop substance use disorders (SUD). Accurate assessment of adolescent AOD use is essential in determining the prevalence of SUDs, the development of effective interventions, and the implementation of beneficial prevention initiatives. Developmental considerations are significant factors in the validity of youth AOD assessment and are detailed in this chapter. PMID- 15789879 TI - Acronyms for the rheumatology literature. PMID- 15789880 TI - Difficulties in the differential diagnosis between primitive rheumatic diseases and hepatitis C virus-related disorders. PMID- 15789881 TI - HFE genotyping demonstrates a significant incidence of hemochromatosis in undifferentiated arthritis. AB - OBJECTIVE: Hereditary hemochromatosis is a common autosomal recessive disorder of iron metabolism. Among Northern Europeans the carrier frequency is estimated to be 1 in 10, while up to 1 in 200 is affected by the disease. Arthropathy is one early clinical manifestation of this disease, but the articular features are often misdiagnosed. In this study the two frequent mutations of the HLA-linked hemochromatosis gene (HFE) were investigated in a rheumatology clinic population. METHODS: Two hundred and six consecutive patients (mean age 57.7 years; 38 male/168 female) attending a rheumatology clinic over a period of 14 months were screened for HFE mutations (C282Y and H63D). All standard diagnostic procedures were used to identify the aetiology of the arthropathy. Mutations were evaluated by separation on PAGE of digested PCR amplificates of DNA (by SnapI and Bcl-I, for C282Y and H63D, respectively) obtained from PBMCs. RESULTS: The C282Y and H63D allele frequencies were 4.5 and 12.8 in patients with rheumatic diseases. Five patients were homozygote for H63D (2.4%), and one for C282Y (0.5%). Five patients were compound heterozygous (2.4%). The observed C282Y allele frequency in rheumatic patients with undifferentiated arthritis was 12.9 and exceeded that of healthy subjects (p = 0.01). CONCLUSIONS: Determination of the HFE genotype is clinically useful in patients with arthritis of unknown origin, to allow early diagnosis of hemochromatosis. PMID- 15789882 TI - Histological features of rheumatoid arthritis patients having antibodies to enterobacterial common antigens: correlation of antibody levels with semiquantitative histologic scores and laboratory markers. AB - OBJECTIVE: To ascertain whether clinically diagnosed rheumatoid arthritis (RA) patients having antibodies to enterobacterial common antigens (ECA) in synovial fluid (SF) have a histological appearance characteristic of RA synovitis. METHODS: Twenty-five RA patients for which synovial biopsy specimens were preserved, were selected from 58 patients with RA tested for antibodies to ECA in SF The synovial tissue specimens were examined histologically using a semiquantitative scoring system with quantitative counts. The correlation of anti ECA antibody levels with total scores for synovitis and laboratory markers in three RA patient groups (markedly positive, moderately to slightly positive, and negative according to anti-ECA antibody levels) was analyzed statistically. RESULTS: Histologic examination in the markedly positive RA group (total score for synovitis, range 18-20 points) revealed typical histological features of rheumatoid synovitis. Total scores for synovitis were significantly higher in both the markedly positive and moderately to slightly positive RA groups than in the negative RA group. A comparison of anti-ECA antibody levels with total scores for synovitis revealed a strong and significant correlation. Furthermore, levels of anti-ECA antibodies were also correlated with rheumatoid factor and C-reactive protein. CONCLUSION: Clinically diagnosed RA patients having anti-ECA antibodies in SF showed typical or characteristic histological features of RA synovitis. Our data suggest that a group of RA patients with an entrobacterial etiology exists in larger groups of patients with RA which is thought to be a heterogeneous disease. PMID- 15789883 TI - Chondrocyte-mediated collagenolysis correlates with cartilage destruction grades in osteoarthritis. AB - OBJECTIVE: Osteoarthritis (OA) is associated with destruction of type II collagen rich hyaline articular cartilage. We hypothesized that classical interstitial collagenases cleave collagen type II, leading to the increased expression of the 3/4 native type II collagen fragment (COL2-3/4C) and the corresponding denatured type II collagen fragment (COL2-3/4M), which could correlate with different cartilage destruction grades. In addition, we assessed whether these fragments could be measured in joint fluid and serve as diagnostic markers. METHODS: Cartilage specimens were obtained from the femoral heads of hip joints from total hip replacement operations. Articular gliding surfaces of the cartilage were categorized into normal (G0), fibrillated (G1), superficiallyfissured (G2) and deeplyfissured (fissures that reach to the subchondral bone) (G3). A histological scoring of the cartilage was also used. COL2-3/4C and COL2-3/4M were detected by immunohistochemical staining. Dot blotting was used to detect these fragments in joint fluid. RESULTS: COL2-3/4C and COL2-3/4M were found in the perichondrocyte matrix around lacunae. Such COL2-3/4C (p < 0.05) and COL2-3/4M (p < 0.05) immunoreactivity was significantly increased in G3 and G2 compared to GO and G1. A positive correlation (n = 35, Spearman rank correlation) was observed between the histological score and the percentage of COL2-3/4C positive lacunae (r = 0.43, p = 0.01) and COL2- 3/4M positive lacunae (r = 0.53, p = 0.001). All 7/7 joint fluid samples contained COL2-3/4C in dot blots whereas only 4/7 contained COL2-3/4M. CONCLUSION: Collagenase-cleaved collagen--both native and denatured- increases as the severity of OA increases, assessed using a macroscopic clinical and microscopic histological grading system. Collagen degradation was always most apparent around chondrocytes. Furthermore, the native COL2-3/4C fragment has potential as a joint fluid marker for OA. PMID- 15789885 TI - Practices for managing a flare of long-standing rheumatoid arthritis: survey among French rheumatologists. AB - OBJECTIVE: To describe the practices of rheumatologists in France for managing a flare in a patient being treated for long-standing rheumatoid arthritis (RA) and to estimate the corresponding costs. METHODS: A survey questionnaire was sent to the 2485 practicing rheumatologists in France; 917 completed questionnaires were returned (37% response rate). The questionnaire collected information on the respondents and on their recommendations for managing a fictional patient with a 10-year history of RA in flare, with a recent episode of neck pain, despite prednisone and methotrexate therapy. Investigational and treatment (first month) costs were estimated from the perspective of society in 2001 Euros. RESULTS: Over 80% of the respondents recommended measuring laboratory inflammation parameters, complete blood cell counts, liver enzymes, serum creatinine, and radiographs (hands, anteroposterior cervical spine view, wrists, knees); 50-70% recommended additional cervical spine incidences, elbow and chest radiographs, and bone absorptiometry. Adding anti-TNF therapy (24%) or another DMARD (10%), increasing the methotrexate dosage (24%), and substituting leflunomide for methotrexate were the main recommended treatments. Most respondents suggested continuing the glucocorticoid in the same dosage (61%) or a higher dosage (36%). Analgesics and non-steroidal anti-inflammatory drugs were recommended by 65% and 41% of respondents and rehabilitation therapy by 83%. The median cost was 500 Euro (mean 1105 Euro; range 80-4089 Euro). CONCLUSION: We found a high level of agreement among French rheumatologists regarding the evaluation of established RA. Marked variations in recommended treatments were observed and translated into major cost differences. PMID- 15789884 TI - The ability of disease modifying antirheumatic drugs to induce and maintain improvement in patients with rheumatoid arthritis. epidemiology of DMARDs treatment in Japan. AB - OBJECTIVE: The effectiveness of the disease-modifying antirheumatic drugs (DMARDs) methotrexate (MTX), bucillamine (BUC), salazosulphapyridine (SASP) and gold sodium thiomalate (GST) over two courses of treatment with a follow-up period of at least 12 months was evaluated in 425 patients with rheumatoid arthritis. METHODS: Clinical efficacy was evaluated on the basis of the numbers of painful and swollen joints, morning stiffness, grip strength, erythrocyte sedimentation rate, C-reactive protein and rheumatoid factor levels before and after treatment. Results were evaluated on the basis of the survival rate (Kaplan Meier method) and the incidence and types of adverse drug reactions (ADR) following single and combined therapies. RESULTS: In the first course of treatment, the survival rates for MTX, GST, BUC and SASP were 52.3%, 40.4%, 33.0% and 24.8%, respectively. The rates of development of ADR were 22.9%, 23.5%, 26.3% and 30.0% for BUC, SASP, GST and MTX, respectively. In the second course, the survival rates for MTX, BUC and SASP were 36.6%, 14.1% and 10%, respectively. CONCLUSION: DMARDs used in the first course of treatment improved the clinical parameters until the 6th month after initiation of treatment. Combination treatments showed some effectiveness, but because of the high incidence of ADR the survival rate was low. DMARDs used in the second course of treatment were not efficacious and there was no improvement in the survival rate compared to the first course of treatment. PMID- 15789886 TI - Hashimoto's thyroiditis is associated with peripheral lymphocyte activation in patients with systemic sclerosis. AB - OBJECTIVE: To investigate whether autoimmune thyroiditis [HT] (i.e., a TH1 disease) influences the pattern of peripheral lymphocyte activation in systemic sclerosis [SSc] (commonly regarded as a TH2 disease). Twenty SSc patients, 6 with (SSc+HT+) and 14 without HT (SSc+HT-) and 20 controls were investigated for the intracellular content of IFN-gamma and IL-4 in unstimulated and stimulated (25 ng/ml PMA and 1 microg/ml ionomycin) CD4+ and CD8+ T lymphocytes. Results Under basal conditions the percentages of CD4+IFN-gamma, CD4+IL-4+ and CD8+IFN gammawere significantly higher in the patients than the control subjects, no significant differences being detectable between the two patient subgroups. Upon PMA stimulation, the 20 SSc patients showed a higher percentage of CD4+IFN-gamma+ and CD8+IFN-gamma+ than the control subjects. In particular, the 14 SSc+HT- patients showed a higher number of CD4+IFN-y+ and CD4+IL-4+ cells, while the SSc+HT+ patients showed higher percentage of CD8+IFN-gamma+ cells. The latter patients showed a reduced percentage of CD4+IL-4+ cells and an increased percentage of CD8+IFN-y+ in comparison with the SSc+HT- patients. Type-1 activation in the peripheral blood of SSc patients has been already pointed out by other authors and ourselves. This study shows that such activation mainly affects SSc patients with coexistent HT. PMID- 15789887 TI - Disease course, frequency of relapses and survival of 73 patients with juvenile or adult dermatomyositis. AB - OBJECTIVE: Our aim is to present the disease course, frequency of relapses and survival of juvenile and adult dermatomyositis (JDM/DM) patients. METHODS: Analysis was performed using data on 73 patients. The median follow-up for 38 JDM patients was 32 months and 78 months for 35 adult DM patients. RESULTS: 23/38 JDM patients (60%) had monophasic, 12/38 (31.6%) had polycyclic and 3/38 (7.9%) had chronic disease. Among children treated only with glucocorticoids, 12/20 (60%) had monophasic and 8/20 (40%) had polycyclic disease. 10/17 (58.8%) children, who required second-line immunosuppressive agents, had monophasic and 4/17 (23.5%) had polycyclic disease. 18/35 DM (51.4%) patients had monophasic, 13/35 (37.1%) had polycyclic, 1/35 (2.9%) had chronic disease and 3/35 (8.6%) had fulminant myositis. Among DM patients requiring only glucocorticoids, 12/20 (60%) were monophasic and 8/20 (40%) were polycyclic. In patients requiring second-line immunosuppressive agents, 6/15 patients (40%) had monophasic and 5/15 (33.3%) had polycyclic disease. Among patients with polycyclic disease, the risk of relapse was higher during first year than later in the disease course. None of the JDM patients have died, while 4 disease-specific deaths occurred in adult patients. There was no significant difference between the survival of JDM and DM patients. DISCUSSION: There was no correlation between relapse-free survival and the initial therapeutic regimen. Many of our patients had polycyclic or chronic disease. As relapses can occur after a prolonged disease-free interval, patients should be followed for at least 2 years. Although we found a favourable survival rate, further investigations are needed to assess functional outcome. PMID- 15789888 TI - Chemokines differentially induce matrix metalloproteinase-3 and prostaglandin E2 in human articular chondrocytes. AB - OBJECTIVE: To explore prostaglandin (PG) E2 production by human articular chondrocytes induced by different chemokines. METHODS: Human chondrocytes were enzymatically isolated from the articular cartilage of patients with rheumatoid arthritis (RA), osteoarthritis (OA) or traumatic fracture (N) who underwent total joint replacement. They were cultured in vitro as monolayers and then exposed to MCP-1, RANTES or SDF-1 for 24 h. Levels of PGE2 and MMP-3 in the culture supernatant were then immunoassayed. RESULTS: PGE2 production was enhanced up to 2.7-fold in a subset of samples. Responses to different chemokines were heterogeneous even within the same disease groups. As previously reported, chemokines induced MMP-3 secretion by chondrocytes, but there was no significant correlation between levels of PGE2 and MMP-3. CONCLUSION: We here document the presence of "responders" among OA, RA and normal chondrocytes that produce enhanced levels of PGE2 upon stimulation by chemokines. The relationship between chemokines and prostaglandins could differentially influence the pathogenic network responsible for cartilage degradation in arthropathy. PMID- 15789889 TI - The effects of Celecoxib on inflammation and synovial microcirculation in murine antigen-induced arthritis. AB - OBJECTIVE: There is controversy about the effects of cyclooxygenase-2 (COX-2) on adhesion molecules and the microvasculature in inflamed tissue. Thus, the aim of this study was to assess COX-2-expression in Antigen-induced Arthritis (AiA) and to investigate the effects of selective COX-2 inhibition by Celecoxib (4-[5-(4 methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide) (CXB), on synovial microcirculation and adhesion molecule expression in arthritic as well as healthy mice. METHODS: Balb/c mice were allocated to 4 groups; 2 control groups with saline or CXB and 2 groups with AiA which also received saline or CXB (30 mg/kg BW in 0.3 ml solution). The severity of arthritis was assessed by changes in the transverse joint diameter On day 14 after AiA-induction, the patella tendon of the left knee joint was microsurgically resected and intravital fluorescence microscopy on synovial tissue was performed. Finally, the knee joint was removed for histology and immunohistochmistry. RESULTS: COX-2-expression in the inflamed synovium was demonstrated by immunohistochemistry. Application of Celecoxib resulted in a significant reduction in the rolling leukocyte fraction as well as in the number of leukocytes adherent to the endothelium (0.25 +/- 0. 1 and 96 +/- 34 cells/mm2 respectively) in comparison to the untreated animals with AiA (0.44 +/- 0.03 and 206 +/- 22 cells/mm2 respectively). Additionally, CXB treated arthritic animals showed significantly less knee joint swelling and reduced adhesion molecule expression. CONCLUSION: In the present study, COX-2 expression in the synovial tissue of mice with AiA could be demonstrated. Selective COX-2 inhibition with CXB resulted in reduced leucocyte-endothelial cell interactions and decreased adhesion molecule expression. Evidence for a protective role of COX-2 in mouse AiA was not found. PMID- 15789890 TI - Reliability, validity and responsiveness of instruments to assess disabilities in personal care in patients with rheumatic disorders. A systematic review. AB - OBJECTIVES: The first aim was to make an inventory of available instruments and questionnaires for the assessment of disabilities in personal care in patients with rheumatic disorders. The second aim was to investigate which of these instruments have acceptable, methodological quality with regard to reliability, validity and responsiveness. The third aim was to investigate the assumption that convergent validity results in stronger correlations when validated against a more similar construct. METHODS: A computer-aided literature search (1982-2001) in several databases was performed to identify studies focusing on the clinimetric properties of instruments to assess impairments in function in patients with rheumatic disorders. Data were extracted in a standardised way and compared to a priori defined criteria. RESULTS: In total, 19 measurement instruments were included. Five out of these 19 were found to have acceptable reliability, while 12 had acceptable validity. Only three questionnaires met both criteria. Results concerning the responsiveness of these three questionnaires were conflicting. No difference was found in the strength of correlation between validation against the most similar construct versus validation against the least similar construct. CONCLUSION: It is concluded that the Arthritis Impact Measurement Scale (AIMS) is the most suitable instrument for the assessment of disabilities in personal care. PMID- 15789891 TI - Effectiveness and safety profile of leflunomide in rheumatoid arthritis: actual practice compared with clinical trials. AB - OBJECTIVE: Leflunomide, an immunosuppressant agent for treating rheumatoid arthritis, was first marketed in France in 2000. Three years after its launch, we sought to assess its prescription patterns in the real world of prescription and use, and to see if its efficacy and safety profiles observed during clinical trials were confirmed. METHODS: All patients treated with leflunomide from May 2000 to April 2003 in the Department of Rheumatology of the Bordeaux University Hospital were identified, and their treatment patterns and outcome ascertained. This was compared to data from clinical trials. RESULTS: 116 were included (mean age = 55 years, 70% women). Almost 21.7% stopped treatment for lack of efficacy (after a mean delay of 3.6 months), 16% for secondary loss of efficacy (median = 7 months), and 32% for the occurrence of an adverse event (half within 4 months). Over a similar time frame in clinical trials, in patients of about the same age and sex but with less severe disease, the corresponding figures were 7-17% for lack or loss of efficacy, and 14-22% for adverse effects. At one year of follow up, the discontinuation rate was 70% in the cohort compared to 28-47% in clinical trials. DISCUSSION: The differences between the two populations confirm the need to conduct post-marketing studies in order to obtain better knowledge on the effectiveness and safety of a new drug. In many cases, a simple drug utilization study can provide relevant information on the degree of shift between populations included in clinical trials and those treated in real life. PMID- 15789892 TI - Interleukin-1 receptor antagonist gene polymorphism in Taiwanese patients with gout. AB - OBJECTIVE: The purpose of this study was to examine whether interleukin-1 receptor antagonist (IL-1Ra) gene polymorphism is a marker of susceptibility to or of the clinical characteristics of gout in Taiwanese patients. METHODS: 196 Taiwanese patients with gout and 103 unrelated normal healthy control subjects living in central Taiwan were studied. Polymorphism of the gene for IL-1Ra was typed from genomic DNA. Allelic frequencies and carriage rates were compared between gout patients and control subjects. The relationship between IL-1Ra genotypes and the clinical characteristics of gout was also evaluated. RESULTS: No significant differences were observed in genetic and allelic frequencies of the IL-1Ra gene polymorphism between patients with gout and healthy control subjects. Furthermore, we did not detect any association of IL-1Ra genotype with the clinical and laboratory profiles in patients with gout. CONCLUSION: The results from the present study suggest that the interleukin-1 receptor antagonist gene polymorphism is not a genetic marker of susceptibility to gout for Taiwanese. Furthermore, our study also suggests that the IL-1Ra gene polymorphism is unrelated to the clinical characteristics of gout. PMID- 15789893 TI - Modulation of cytokine release by purine receptors in patients with rheumatoid arthritis. AB - OBJECTIVE: Since adenosine receptors are known to modulate the release of some inflammatory mediators in control subjects, we have examined the effects of the mixed A1 and A2 adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA) on basal and lipopolysaccharide (LPS)-induced cytokine release in diluted whole blood cultures from rheumatoid arthritis (RA) patients and healthy volunteers. METHODS: Twenty-eight patients with rheumatoid arthritis aged 18-75 years gave their voluntary consent to participate and give a blood sample. Basal levels of tumour necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) were measured by ELISA, and whole blood cultures were prepared to assess the effects of LPS activation. RESULTS: Following a 40-hour incubation, activation of adenosine receptors by NECA, added to the cell cultures from rheumatoid arthritis patients, was found to suppress both the basal and LPS induced release of TNF-alpha and IL-1beta, while causing an increase in the release of both basal and LPS-induced IL-6. In healthy volunteers basal cytokines were undetectable, but NECA alone induced the release of all three cytokines. Stimulated levels of TNF-alpha were more than double those in patients. In the control blood cultures, NECA suppressed LPS-induced release of TNF-alpha and IL 1beta, but increased IL-6 release. CONCLUSIONS: Adenosine receptor stimulation has a differential effect on the release of pro-inflammatory cytokines, and may induce cytokine release in normal subjects. Stimulated release of TNF-alpha is substantially lower in patients with rheumatoid arthritis than in control subjects, possibly indicating saturation, exhaustion or down-regulation of the release process. PMID- 15789894 TI - Steroid-responsive inclusion body myositis associated with endometrial cancer. AB - Inclusion body myositis (IBM) is an uncommon chronic inflammatory myopathy. Although the association between other myopathies and cancer has been well established, the relationship between IBM and neoplasia is not completely understood. Unlike polymyositis (PM) or dermatomyositis (DM), IBM rarely responds to immunosuppressive treatment and the response is seldom long-lasting. We describe a case of IBM associated with endometrial carcinoma that also demonstrated a unique response to steroids alone which persisted despite cancer relapse. The factors that are associated with a response of IBM to steroids are discussed. An atypical, steroid-responsive form of the disease is delineated. PMID- 15789895 TI - Refractory multicentric reticulohistiocytosis treated by infliximab: two cases. AB - We report the effect of infliximab, a monoclonal anti-TNFalpha antibody, in two patients with refractory cutaneous and articular multicentric reticulohistiocytosis (MRH). One 37-year-old woman and one 53-year-old woman with polyarthritis, facial rash and nodular lesions on the hands related to MRH were refractory to multiple agents: cariolysine, corticosteroids, hydroxychloroquine and cytotoxic agents. Infliximab at 3 mg/kg which was then increased to 5 mg/kg in combination with methotrexate or azathioprine was effective on cutaneous manifestations of the disease but not on polyarthritis. A switch to etanercept did not improve polyarthritis in the second patient. Some data suggest that TNFalpha is involved in MRH, but based on our cases anti-TNFalpha therapy needs further evaluation in patients with refractory MRH. PMID- 15789896 TI - Cytomegalovirus-associated hemophagocytic syndrome in a patient with adult onset Still's disease. AB - Reactive hemophagocytic syndrome (HPS) is characterized by hemophagocytosis by activated histiocytes, resulting in pancytopenia and liver dysfunction. We describe a patient with adult onset Still's disease (AOSD) in whom HPS developed. An 80-year-old Japanese woman with high fever, arthralgia, skin rash, and pleuritis was admitted to our hospital for further examination. She was diagnosed with AOSD and steroid therapy was initiated. During the course of steroid therapy, a re-elevation of serum ferritin levels and a marked increase in serum transaminase were observed. Bone marrow aspiration revealed an increase in the number of histiocytes with hemophagocytosis and cytomegalovirus (CMV)-positive leukocytes were detected. At this time we diagnosed the patient as having virus associated hemophagocytic syndrome (VAHS) and elevated levels of trasaminase and ferritin were normalized by ganciclovir treatment. Reactive HPS occurs in cases of active AOSD. However, it should be noted that HPS may be accompanied by opportunistic infections during immunosuppressive therapy requiring prompt antibiotic therapy. PMID- 15789897 TI - Overview of studies comparing human normal cartilage with minimal and advanced osteoarthritic cartilage. AB - A major area under study in the osteoarthritis (OA) research field is the characterization of specific molecular and biochemical changes that distinguish advanced diseased cartilage from less involved or normal tissue. This information is important to better define the pathogenic mechanisms that are operating during OA progression and to identify disease-specific markers. This review describes recent studies that have addressed changes in chondrocyte gene expression, proliferation, and apoptosis in "experimental" (more advanced OA cartilage) versus "control" (less involved or non-OA cartilage). Included is a comprehensive listing of recently published studies in this area with general findings. The review also includes a discussion of study design and the strengths and weaknesses of the various approaches. In addition, specific strategies to deal with some of the important issues are discussed. One particular model utilizing minimal and advanced OA cartilage obtained from the same patient is described in more detail. PMID- 15789898 TI - Inter-observer agreement of the physician's global assessment of disease activity in children with juvenile idiopathic arthritis. AB - OBJECTIVE: To assess the inter-observer agreement in the physician's global assessment of overall disease activity (MD global) in a cohort of patients with juvenile idiopathic arthritis (JIA). METHODS: Forty consecutive patients with JIA, who were representative of a wide spectrum of disease activity and severity, were examined simultaneously by 4 observers. Observer 1 (who was the most experienced rheumatologist) carried out a routine rheumatologic examination of each patient including a complete articular assessment, and subsequently calculated in secrecy the MD global score on an anchored horizontal 10-cm VAS. Observers 2, 3 and 4 were present during the examination; afterwards they also scored in secrecy the MD global score for the patient. Agreement was measured by the intra-class correlation coefficient (ICC), using the score of Observer 1 as the gold standard. An ICC below 0.75 was considered unsatisfactory. RESULTS: The mean (SD) MD global scores for Observers 1, 2, 3, and 4 were 5.2 (3.4), 6.7 (3.9), 5.9 (3.5), and 5.6 (3.7), respectively. The level of agreement with Observer 1 in scoring was 0.83 for Observer 2, 0.88 for Observer 3, and 0.90 for Observer 4, indicating good agreement for all observers. CONCLUSIONS: Our study shows a good inter-observer agreement in the physician's global assessment of overall disease activity in patients with JIA. Analyses involving investigators from different countries are needed to determine whether these results can be generalized. PMID- 15789899 TI - DRB1, DQA1, DQB1 genes in Turkish children with rheumatic fever. AB - OBJECTIVES: Several studies have suggested that genetic susceptibility to rheumatic fever (RF) may be linked to HLA Class II alleles. The purpose of this study was to examine the association between HLA Class II genes and RF in Turkish children. METHODS: DNA typing HLA Class II genes (DRB1, DQA1, DQB1) were performed in 55 children with RF and 50 healthy unrelated controls using sequence specific primers (SSP). RESULTS: The frequency of the HLA DQA1*03 (OR: 0.462, p < 0.05) allele was significantly decreased in the patient group. Also, the frequency of the combination of DRB1*04 and DQA1*03 allele (OR: 0.42, p < 0.01) was more significantly decreased in the patient group. Differences in frequencies of the DRB1 and DQB1 alleles between groups were not significant. CONCLUSIONS: Our data indicate that the HLA DQA1*03 allele may be a protecting factor in Turkish children with RF. Our results also suggest that the combination of the DRB1*04 and DQA1*03 alleles may be a stronger protective factor than the DQA1*03 allele alone. PMID- 15789900 TI - Listeria monocytogenes meningitis in a patient receiving etanercept for Still's disease. PMID- 15789901 TI - Prolonged efficacy of infliximab for refractory adult-onset Still's disease. PMID- 15789903 TI - Prevalence of diffuse idiopathic skeletal hyperostosis in a female Italian population. PMID- 15789902 TI - Late onset of long-lasting fever as a sole complication of treatment with anti TNFalpha. PMID- 15789904 TI - Intestinal perforation and jejunal haemorrhage due to Wegener's granulomatosis. PMID- 15789905 TI - Circulating eosinophils lack ex vivo chemotaxis toward vascular endothelial growth factor in a patient with Churg-Strauss syndrome. PMID- 15789906 TI - Thyroid acropachy: an unusual rheumatic manifestation of Graves' disease. PMID- 15789907 TI - Calcinosis universalis in systemic sclerosis with limited scleroderma. PMID- 15789908 TI - [Possibilities of MR evaluation in thyroid orbitopathy]. AB - Thyroid orbitopathy is one of the main indications for orbital imaging. MR yields most complex, both quantitative and qualitative information concerning extraocular muscle pathology, including eye motility disorders. This examination is helpful in the treatment planning and evaluation of its efficiency. PMID- 15789909 TI - [Nuclear medicine imaging methods in thyroid associated ophthalmopathy]. AB - The evaluation of inflammation activity is one of the more important problems in the diagnosis of thyroid associated ophthalmopathy (TAO). It is connected with response to immunosuppresive treatment. Some of the radiolabelled tracers used in nuclear medicine are collected selectively in inflammatory foci and are also accumulated in inflammable retrobulbar tissue present in the active phase of TAO. This enables an evaluation of the inflammatory process activity and concerns especially such tracers as OctreoScan, Ga-67, Tc-99m-DTPA. This article presents a short review of current knowledge on the role of nuclear medicine methods in thyroid associated ophthalmopathy imaging. PMID- 15789910 TI - [Difficulties in diagnosing Graves' ophthalmopathy--case report]. AB - PURPOSE: The aim is to present diagnostic difficulties which may occur when diagnosing Graves' ophthalmopathy. MATERIAL AND METHODS: 49-year-old woman, who had been smoking 20 cigarettes a day since she was 21 years old, treated for Graves' disease for 14 years, with the right eyeball protrusion existing for 6 months, dysfunction of eye movements and diplopia, was admitted to the Diabetes and Endocrinology Center in Bydgoszcz. After ophthalmological examination the patient underwent MRI and USG of the orbits which showed a change in the inferior rectus muscle of the right eye which could correspond to orbit tumor. In the Ophthalmology Department of the Medical Academy in Bydgoszcz the opening of the orbit was carried out in order to confirm or to eliminate the neoplastic process. RESULTS: Surgery and control examinations excluded neoplastic process. CONCLUSION: It is very difficult to differentiate between inflammatory infiltration and neoplastic tumor in patients with unilateral protrusion in the course of Graves' ophthalmopathy. PMID- 15789911 TI - [The endocrinological management of thyroid associated orbitopathy in relation to severity and activity of the disease]. AB - The complete diagnosis of thyroid associated orbitopathy (TAO) comprises endocrinological and ophthalmological assessment of patients. This approach facilitates classification of patients and qualification of patients for the proper treatment of eye changes. The aim can be obtained by assessment of severity and activity of the disease. One can distinguish at least 4 clinical situations (1. severe and active TAO, 2. severe and inactive TAO, 3. non-severe and active TAO, 4. non-severe and non-active TAO). Methods of reestablishing euthyreosis are proposed in this article, in relation to disease activity and severity of the disease. The two case-reports are described in order to express proposed theses. PMID- 15789912 TI - [The effect of one-month antithyroid therapy on peripheral metabolism of reactive oxygen species in Graves' disease with infiltrative ophthalmopathy]. AB - Local inflammation is characterized by oxidative stress present in situ, and may also influence reactive oxygen species peripheral metabolism. Infiltrative ophthalmopathy is considered as an inflammatory disorder of autoimmune background. This study estimated influence of hyperthyroidism and antithyroid treatment on peripheral parameters of oxidative stress in 65 Graves' patients (40 with and 24 without infiltrative ophthalmopathy). Age- and sex-matched 25 healthy volunteers served as controls. In all hyperthyroid patients hydrogen peroxide (H2O2), lipid hydroperoxides (ROOH), malondialdehyde, ceruloplasmin (CP), superoxide dismutase (SOD) and catalase (CAT) were increased, whereas glutathione peroxidase (GPx) and glutathione reductase (GR) were reduced. One-month methimazole treatment produced further elevation in H2O2 in all patients and CAT in the ophthalmopathy group, as well as partial reversal of SOD and CP in all patients, ROOH, GPx and GR in the non-ophthalmopathy group and CAT in patients without ophthalmopathy. In summary, our results confirm that changes of blood extracellular indices of reactive oxygen species metabolism in Graves' disease are influenced by the thyroid metabolic status. However, the differences of the parameters analysed after achievement of euthyroidism in the patients with and without infiltrative ophthalmopathy suggest an involvement of factors, presumably connected with orbital inflammation, which modify oxidative stress intensity. PMID- 15789913 TI - [Lanreotide in the treatment of thyroid orbitopathy]. AB - The aim of the present study was to evaluate the efficiency of somatostatin analogue--lanreotide in the treatment of Graves ophthalmopathy. We examined the group of 6 women with thyroid eye disease in class 2 and 3 according to NO SPECS classification. Ophthalmopathy index and clinical activity score were above 3. Lanreotide was given in a dose of 30 mg i.m. every 2 weeks over a period of 3 months. The therapeutic effect was evaluated twice: 2 weeks after the end of lanreotide treatment (initial assessment) and after 2 years (final assessment). On initial assessment a decrease in ophthalmopathy index more than 2 points in each eye was observed. On follow up after 2 years we did not observe the worsening of thyroid eye disease. IN CONCLUSION: lanreotide is an effective drug in reduction of soft tissue inflammatory changes in patients with active Graves ophthalmopathy. Absence of recurrence of the disease in final assessment indicates the persistent effect of lanreotide treatment. PMID- 15789914 TI - [Surgical management of thyroid orbitopathy]. AB - Thyroid orbitopathy (TO) requires often not only pharmacological treatment but also surgical procedures. The management contains orbital decompression (wall decompression, excision of fat), correction of lid retraction and strabismus operations on extraocular muscles. Authors describe indications for surgical procedures, methods of operations, results and complications of own material of 99 patients with TO treated in the Eye Clinic in Poznan and Plastic Surgery and Eye Clinic in Kobylniki from 1999 until 2003. PMID- 15789915 TI - [Effectiveness of treatment for thyroid orbitopathy in patients hospitalized at the Endocrinology Department of Pomeranian Medical University]. AB - The aim of the present study is the assessment of efficacy of systemic corticosteroid treatment combined with orbital radiotherapy in patients with thyroid ophthalmopathy. MATERIAL: 101 patients hospitalized in the Endocrinology Department of the Pomeranian Medical University in the years 1999-2002. METHODS: During first stage of treatment 3 to 6-7 intravenous pulses of methylprednisolone (SoluMedrol Pharmacia & Upjohn) in a dose of 1.0 g/day in consecutive 3 days were administered in combination with radiotherapy of the retroorbital spaces. Ten daily doses of 200 cGy using X-ray to each retroorbital space were given between the 2-nd to 4-th SoluMedrol pulse. Prednisone (Encorton Polfa) at an initial dose of 60 mg/day was used during the second stage of therapy. RESULTS: Donaldson's ophthalmopathy index scored for each eye separately decreased significantly after both stages of treatment (right eye: 6.35 (before treatment); 2.25 (after first stage), 1.2 (after second stage), p<0.05; left eye: 6.1 (before treatment); 2.1 (after first stage); 1.15 (after second stage), p<0.05). The clinical picture of Graves ophthalmopahty after the end of therapy in most of patients was significantly improved. Persistent diplopia was found in 16/101 patients. Two of them were referred for surgical correction of abnormal eyeball fixation, the rest were using prismatic glasses. Until now, no recurrence of thyroid eye disease in treated patients was observed. CONCLUSIONS: Combined therapy is an effective method of treatment of Graves' ophtahlmopathy in terms of both rapid regression of inflammatory changes in orbital soft tissues and in prevention of recurrence of the disease. PMID- 15789916 TI - [Complications during systemic corticosteroid therapy combined with orbital radiotherapy in patients with graves orbitopathy]. AB - OBJECTIVE: An assessment of complications during systemic corticosteroid therapy combined with orbital radiotherapy in patients with thyroid eye disease. MATERIAL: 101 patients including 86 women and 15 men hospitalized in the Endocrinology Department of Pomeranian Medical Academy in 1999-2002. METHODS: During the first stage of therapy, methylprednisolone in a daily dose of 1.0 g was administered intravenously for the following 3 days. The number of methylprednisolone pulses ranged from 3 up to 6-7. Orbital radiotherapy was usually performed between 2 and 4 pulse. Ten daily doses of 200 cGy were given in 2 weeks. After the series of methylprednisone and orbital radiotherapy had been finished, patients were treated with prednisone (Encorton, Polfa) given orally with an initial dose of 60 mg/day. RESULTS: During the first stage of therapy carbohydrate metabolism disturbances were observed in 27 patients. Cardio vascular complications were noted in 8 patients. Four patients complained of digestive tract dysfunction. Acute psychosis was diagnosed in 3 patients. Twelve patients developed infectious diseases. During prednisone administration, 1 case of bacterial pneumonia, 1 purulent meningitis and 1 schizophrenia onset were observed. In prolonged ophthalmologic follow-up 3/101 patients with cataract needed surgical treatment. PMID- 15789917 TI - [Quality of life in Graves' ophthalmopathy (continuation of research)]. AB - The previous research concerning "quality of life" in Graves patients with ophthalmopathy appeared to be a very important direction in health promotion. This drives us to continue enhancement of the quality of life in this group. The biggest deficiency we found to be in the life sphere activity and professional activity, interpersonal relations and positive self-estimation. We analyzed 48 women with Graves' Ophthalmopathy from 32 to 64 yr., average 43 SD 8,17. We used the following experimental methods: Quality of Life Experimental Inventory, Temperament Inventory (Eas-D). In Addition for Adults, STAI Questionnaire (State Trait Anxiety Inventory) and Beck's Depression Scale. We interpreted obtained results as: 1. anxiety tendency, 2. subjective decreased mood, 3. decreased social contact tendency. PMID- 15789918 TI - [Differentiated thyroid cancers. Etiopathogenic and clinical data, prognostic factors and surgical treatment]. AB - Thyroid cancer is the most common among endocrine malignant neoplasm. The cause of its grow is as well the external factors influence and primary malfunction of thyroid cells. The most common is papillary cancer but in the regions where iodine level is low the incidence of follicular cancer increases. The postoperative procedures depend on the type and stage of cancer. Nowadays centers which deal with thyroid cancer treatment are for primary radical surgery as it is followed by lower possibility of missing the cancer in the thyroid remnant, easier postoperative follow up and supplementary treatment management. PMID- 15789919 TI - [Clinical implications of occurrence of antithyroid antibodies in pregnant women and in the postpartum period]. AB - Antithyroid antibodies are classified to immunoglobulin G. It is a varied group of antibodies as there are antibodies against TSH-receptor, against thyroid peroxidase and also against thyroglobulin. Pregnancy is a period in which the titres of antibodies decrease to protect fetus from abortion; but just after delivery they increase again. The clinical implications of this fact are varied and concern not only the thyroid gland but also other organs. Postpartum thyroid dysfunction (PPD) is one,possible disturbance due to presence of antithyroid antibodies. It can be divided into two various types: a) postpartum thyroiditis, b) Graves'-Basedow disease after delivery. Postpartum thyroiditis (PPT) is an example of autoimmune disease connected with many different factors such as genetic or environmental, but the most important factor is the presence of antibodies against thyroid peroxidase. PPT occurs in 50% of women with high titre of these antibodies. Higher risk of PPT also occurs within women with type I diabetes mellitus in comparison with the population, as well as within women smokers. It is also proved that women with high titres of antibodies against TSH receptor are more likely to suffer from Graves'-Basedow disease after delivery. The pathogenesis of postpartum depression is multifactorial. The occurrence of stressful life events (marital disharmony, housing and socioeconomic problems) and some biological factors (e.g. previous psychiatric illnesses) are strongly associated with postpartum depression. Some authors also said that postpartum depression depends on the presence of antithyroid antibodies during pregnancy. It is believed that cytokines which are released during the autoimmune process can affect the central nervous system and can determine changes in behavior. Some authors suggest that changes in concentration of thyroid hormones during the natural history of PPT can be connected with depression after delivery. It is also reported that high titres of antithyroid antibodies are linked with pregnancy loss but the results are not uniform. PMID- 15789920 TI - [Assessment of early immunosuppressive therapy in the prevention of complications of Graves' disease]. AB - Regardless the autoimmune origin of Graves' disease, the preferred method of its treatment remains antithyroid drug administration. Use of immunosuppressive agents (mostly steroids) is still limited to the therapy of disease complications, such as proliferative ophthalmopathy. The aim of the study was to assess the influence of early immunosuppressive treatment of autoimmune thyrotoxicosis with azathioprine on the course of the disease and the incidence of its complications. The study comprised 64 patients (47 females and 17 males aged 20-43 years) for the first time diagnosed with Graves' disease. The subjects were randomised into two groups. Group I consisted of 28 patients treated only with antithyroid drugs, the remaining 36 subjects additionally receiving azathioprine were included into group II. The dose of both drugs was adjusted during the treatment according to metabolic status of each patients. The treatment was continued for 8-14 months, the follow-up duration after therapy withdrawal was 5 years. Euthyreosis was achieved in all patients 2-8 weeks after treatment initiation. No drug intolerance symptoms were observed in group I. In four patients additionally treated with azathioprine, gastrointestinal side effects or leucopenia were present. The disease relapse was observed during the follow-up period in 15 (53.5%) patients of group I and in 3 (8.3%) of group II, the difference was statistically significant (p<0.01). Only one patient receiving additionally azathioprine presented ophthalmic symptoms compared with seven subjects (25%) treated only with antithyroid drugs (p<0.001). The patients of group I were also more frequently referred to surgical treatment due to rapid goitre growth (accordingly 5 (17.8%) and 1 (2.7%) patients, p=0.07), the difference between both groups not being statistically significant. CONCLUSIONS: Additional early immunosuppressive treatment significantly decreased frequency of Graves' disease complications and thyrotoxicosis recurrence. The use of azathioprine may be advised in patients with contraindications to the radical Graves' disease treatment and in prophylaxis of its complications. PMID- 15789921 TI - [Incidence of Graves disease in children in some regions of south-eastern Poland]. AB - The aim of the studies was the assessment of Graves disease (G-D) incidence in children aged 0-15 years who inhabited the Krakowskie (K), Nowosadeckie (N) and Rzeszowskie (Rz) provinces in the years 1989-1996. The incidence of G-D was evaluated depending on the age, sex and place of residence (urban vs. rural area). The analyses for particular years were carried out separately for each province and for three provinces combined. The mean incidence coefficient (I.C.) for three provinces in the years 1989-1996 was 1.83. The highest incidence of G-D was noted in the group of 10-15 year old patients (I.C.=3.81). In the youngest children the disease was sporadic (I.C.=0.12). A significantly higher incidence rate was observed in girls, in whom the I.C. value was the highest in the oldest age group, amounting to 6.92. In boys the incidence of G-D was 8 times lower and the I.C. value in the 10-15 year group was 0.79. It was also demonstrated that the incidence of Graves disease in the entire group showed no significant differences in urban and rural areas. The incidence of Graves disease in subsequent years of the analyzed period was diversified. The linear regression analysis demonstrated a growing tendency in the years 1989-1996. In the period under investigation, the highest incidence was noted in the Rz region, where the standardize incident coefficient was 2.4. In the K and N provinces the incidence was lower and both cases amounted to 1.6 after standardization. The incidence of Graves disease showed a seasonal variation, associated with a significantly more frequent diagnosis established in autumn months. PMID- 15789923 TI - [Oral cavity health status among dentistry students in the Medical University of Bialystok]. AB - The aim of this study was to analyse oral cavity hygiene, general health status and health behaviour of students studying on the last two years on the faculty of dentistry in the Medical University of Bialystok. 127 students were examined: 48 (37.8%) from the fourth and 79 (62.2%) from the fifth year of schooling. We used a standardized and anonymous questionnaire that was made in Collegium Medicum of the Jagiellonian University in Cracow. Obtained data were statistically elaborated. This work was a part of a multicenter study. 41 (32.3%) students suffered from chronic systemic diseases especially allergy (22.8%). 19 (15.1%) of them still undergo drug treatment. As for dental examination, it have been noted that caries prevention was unsatisfactory at early stages of life of the respondents. Oral hygiene was correct. University education has a bearing on quality of students' health behaviour. Parodontium status was adequate. High frequency of temporomandibular joint pathology is alarming and strongly suggests initiating diagnostics and treatment. PMID- 15789922 TI - [Assessment of possible correlations between endogenous androgens and the extent of coronary heart disease and left ventricle function]. AB - In men, androgens and especially testosterone are considered responsible for the much higher rate of coronary artery disease. The male gender is an independent coronary artery disease risk factor. An adverse correlation between endogenous testosterone levels and the extent of coronary atherosclerosis has been demonstrated in just one study. In our study, we investigated the associations between endogenous sex hormone levels and the extent of coronary atherosclerosis, ejection fraction of the left ventricle and coronary heart disease risk factors. PMID- 15789924 TI - [Anesthesia of a patient with recent cardiac infarct for the implantation of a bifurcated aorto-femoral prosthesis. Case report]. AB - The case of 44-year old patient anaesthetized in an emergency course of action for implantation of a bifurcated aorto-femoral prosthesis (Leriche syndrome), with an accompanying acute rest pain in both lower limbs. Three months earlier patient experienced a cardiac infarct complicated by cardiogenic shock and circulatory arrest. Described case proves that: cooperation of an anesthesiologist, cardiologist and surgeon during preparation for surgery on the abdominal aorta in a patient with previous myocardial infarct, selection of an anesthetic as well as postoperative treatment in the intensive care unit, reduce risk of postoperative cardiologic complications. PMID- 15789925 TI - Obesity on the rise. Leads to workplace challenges, patient concerns. PMID- 15789926 TI - Who, why, what and how? PMID- 15789927 TI - Are you a "yes person"? A question of ethics. PMID- 15789929 TI - Assessing dentists' well being. PMID- 15789928 TI - Disability insurance. PMID- 15789930 TI - The new appeal of dividend income. PMID- 15789932 TI - Time tested and still standing tall: a conversation with senator Gerald Cardinale [interview by Eric R. Elmore]. PMID- 15789931 TI - Help patients achieve whiter, brighter smiles and save their lives in the process. PMID- 15789933 TI - So... have you ever heard of something called Artglass? PMID- 15789934 TI - Infection control corner--sterilization: a review. PMID- 15789935 TI - People of New Jersey rank 19th nationally in health & social standings. PMID- 15789936 TI - Change in perceived health and functioning over time in patients with type I diabetes mellitus. AB - OBJECTIVE: The objective of this study was to investigate perceived health and functioning (PHF) of patients with type I diabetes mellitus (DMT1) over time and to compare change in perceived PHF with that of a sample of the general population. METHODS: In a Dutch cohort of 234 patients with DMT1 we assessed PHF over time with two generic instruments: RAND-36 and EuroQol. We applied multilevel modelling to estimate change in PHF over the years 1995--2001. We compared change in PHF with change estimated from a comparably aged sample from the general population using a one-sample t-test. RESULTS: Patients reported a significant decrease in PHF for most RAND-36 subscales. Mean changes in RAND-36 scores ranged from -0.09 (mental health) to -1.18 per year (bodily pain). EQ scores decreased significantly as well. Patients in the cohort had a faster decrease in three RAND-36 subscales and EQ-VAS over time than the estimated decrease in the general population. CONCLUSION: This study showed that patients with DMT1 have a faster decrease in PHF over time than comparably aged persons from the general population. The generic instruments used in our study were sensitive enough to measure changes in PHF over time in an adult diabetes population. PMID- 15789937 TI - Subjective quality of life in patients with chronic fatigue syndrome. AB - The aim of this study was to (1) assess Subjective Quality of Life (SQOL) of patients with Chronic Fatigue Syndrome (CFS) using a generic concept and to compare the findings with those in groups with mental disorders and healthy subjects, and (2) investigate whether and, if so, to what extent socio demographic and clinical variables predict SQOL in CFS patients. Seventy-three patients diagnosed with CFS were randomly selected and interviewed from two specialised clinics. CFS was diagnosed using the Oxford Criteria. SQOL was assessed on the Manchester Short Assessment of Quality of Life (MANSA) and Health Related Quality of Life (HRQOL) on the Medical Outcome Study Short-Form 36 (MOS) SF-36. A battery of mood and symptom questionnaires, including the Symptom Checklist Questionnaire (SCL-90-R), was administered to assess various aspects of symptomatology as potential predictor variables. Multiple regression analyses were conducted to identify predictors of SQOL. Overall, SQOL was low in CFS patients and less favourable than in groups with mental disorders and healthy subjects. Satisfaction was particularly low with life as a whole, leisure activities and financial situation. Whilst SQOL was only moderately correlated with HRQOL, the SCL-90-R score, especially SCL-90-R Depression scale score, was the best predictor of SQOL explaining 35% of the variance. HRQOL and generic SQOL appear distinct despite some overlap. The findings underline that SQOL is significantly disrupted in CFS patients. Depressive symptoms are statistically the strongest 'predictor' of SQOL, although the direction of the relationship is not established. These data suggest that treatment of depression associated with CFS, regardless of causation, could help to improve SQOL in CFS patients. PMID- 15789938 TI - Patient reported outcomes in gastroesophageal reflux disease: an overview of available measures. AB - Gastroesophageal reflux disease (GERD) is a common, chronic disorder. The main symptom of GERD is heartburn, although a diverse range of symptoms can be associated with the disease including acid regurgitation and epigastric pain. GERD is also a risk factor for Barrett's oesophagus and esophageal adenocarcinoma. The impact of GERD symptoms on patients' lives can be profound and is unrelated to the presence or absence of esophagitis. The impact of GERD can be measured by assessing the patient perspective using Patient Reported Outcomes (PROs). There are two categories of questionnaires that can be used to measure the effect of GERD on health-related quality of life (HRQoL), namely generic and disease or treatment specific. The use of PRO instruments has become more accepted in the assessment of disease treatment. Well-designed instruments that assess physical, psychological and emotional factors can provide clinicians with the data that will promote effective management decisions for the treatment of GERD. The most frequently used instruments in GERD are reviewed here, in terms of their psychometric properties. PMID- 15789939 TI - Determinants of the quality of life of patients with congenital heart disease. AB - OBJECTIVE: The improvement of the quality of life of chronically ill patients has become an important goal in treatment. However, it is seldom taken into account that many other factors, in addition to somatic factors, have an influence on the quality of life of patients. Using patients with congenital heart defects as an example, we examined the relative significance of biological factors, compared to psychological and social factors, for the various quality of life dimensions. RESEARCH DESIGN AND METHODS: One hundred and eleven patients (aged 33+/-12 years) with different degrees of cardiac dysfunction were examined (NYHA 0: 2 I: 56, II: 38, III: 13, IV: 2). All patients for whom there was no contra-indication underwent a treadmill ergometry in order to determine their level of cardiopulmonary functioning (peak oxygen consumption: VO2max). All patients were asked to fill out questionnaires concerning their quality of life (WHOQOL-Bref), their cardiac complaints (Giessener Complaint Questionnaire GBB), their personality traits (Giessen Test GTS), and the social support they experience (Social Support Questionnaire SOZU-k22). The data were analyzed using a linear structural equation model (SEM). RESULTS: In all aspects but the social domain, the HRQL of CgHD patients was significantly diminished compared to the normal population. The SEM proposed was valid, showing good indices of fit (chi2 = 1.18; p = 0.55; AGFI = 0.92). The level of cardiopulmonary functioning was most significant for the reporting of specific cardiac complaints (beta = -28) and for the physical component of the general HRQL (beta = 32), although the former was also influenced by a depressed disposition (beta = -0.20) and the extent of social support experienced (beta = 0.18). The objective findings, however, had virtually no individual significance for the psychological (beta = 0.09) and social domains (beta = -0.02). These HRQL domains are primarily influenced by depressive personality traits (beta= -26/-0.16) and the social support experienced (beta = 0.51/0.51). CONCLUSIONS: The patient's organic dysfunction primarily determines illness-specific complaints but has little relevance for the psychological and social aspects of the HRQL. These aspects are predominantly determined by the patient's depressive disposition and by the experienced social support. A successful therapy should therefore take biological as well as psycho social determinants of the quality of life into account. PMID- 15789940 TI - Comorbidity of chronic diseases: effects of disease pairs on physical and mental functioning. AB - Although much research has been done on the impact of chronic illness on quality of life, still relatively little is known about the role of comorbidity. Given the growing number of (older) people with multiple chronic conditions, more information is needed on the effects of specific disease combinations for preventive purposes. In a nationwide representative sample of 1673 non institutionalized chronic disease patients (recruited in 56 general practices) in The Netherlands, we assessed the separate and joint effects of cardiovascular disease, cancer, arthritis, chronic respiratory disease, diabetes mellitus, and thyroid dysfunction on physical and mental functioning. Data on medical diagnoses were provided by the general practitioners; data on physical and mental functioning were collected by a patient survey (SF-36). Compared to reference data of the general population, physical functioning appeared to be worse in all six diagnostic groups, whereas mental functioning was more or less comparable. Patients with arthritis or those suffering from comorbidity reported the lowest levels of physical functioning. Synergistic effects of combinations of diabetes, cardiovascular disease and/or chronic respiratory disease were found, indicating that patients suffering from these disease combinations run a higher risk of physical disability than could be expected from their separate effects. PMID- 15789941 TI - Perspectives on quality of life of people with intellectual disabilities: the interpretation of discrepancies between clients and caregivers. AB - Large discrepancies have sometimes been found between the quality of life (QOL) experienced by clients with intellectual disabilities and their QOL as described by their caregivers. Olsen and Schober (Soc Indi Res 1993; 28: 173-193) may have provided a framework useful to conceptualize such discrepancies. They suggested that lack of concordance between objective circumstances (i.e. poverty) and perceived QOL must not be treated as measurement noise, but as a source of information. They speculated on the psychological processes that underly and explain this lack of concordance; particularly processes that try to deal with the stress induced by cognitive dissonance and learned helplessness. In this study low to moderate agreement was found between caregivers judgement of QOL of individual clients and clients' own perception of QOL. In some areas and for some clients the discrepancies indicated dissonance (caregivers are satisfied, clients are not) and in other areas and for other clients adaptation (clients are satisfied, caregivers are not). There were systematic differences between clients falling in these groups with respect to their personal characteristics and with respect to factual conditions of care. These outcomes are interpreted with respect to Olson and Schober's contentions regarding the psychological processes that underly a person's experience of QOL. The Olson and Schober framework appears to be a fruitful way to combine different sources of information regarding QOL. PMID- 15789942 TI - Psychometric properties of the general well-being index. AB - There is a growing interest in the measurement of quality of life as an important marker of health outcomes. The General Well-Being Index (GWBI) is a quality of life measure that has been specifically designed to assess psychological distress rather than physical incapacitation. Previous studies using the GWBI have mainly focussed on clinical samples (e.g., depressed patients). The aim of the current study was to examine the psychometric properties of the GWBI in a non-clinical population. Using data gathered from 447 first year University students, the GWBI was found to have excellent psychometric properties. Both internal consistency and test-retest reliability were high, and the concurrent and construct validity of the measure were sound. Factor analysis revealed three significant factors, which were labelled 'general mood/affect', 'life satisfaction/vitality', and 'poor physical health/somatic complaints'. Limitations to the present study are further discussed. PMID- 15789943 TI - Development, feasibility and compliance of a web-based system for very frequent QOL and symptom home self-assessment after hematopoietic stem cell transplantation. AB - We believe that many adverse events following hematopoietic stem cell transplantation (HSCT), particularly relapse and chronic graft-versus-host disease (CGVHD), are preceded by a subclinical period of development that is accessible by frequent psychometric assessment. Documenting these associations could improve future clinical care by extending the potential window for intervention. However, conventional methods of assessing quantity of lite (QOL) in patients in their homes, typically by mailed self-assessment questionnaires, are impractical for very frequent administration. We have developed and implemented a web-based system for measuring short-term (dynamic) changes in QOL by employing brief, online, daily QOL assessments and more extensive, monthly online assessments from patients' homes. Here we report the feasibility of collecting very frequent patient home self-assessments of QOL via the web for a 52 week participation period; we detail incidence of home web access, accrual, compliance, and satisfaction with the system in an HSCT patient sample. We also describe our integrated web-systems for administering patient recruitment, scheduling, monitoring, and analysis. Our results suggest that very frequent routine collection of QOL outcomes is entirely feasible using our web-based home assessment tool, with good patient compliance and high user satisfaction. We believe our methodology shows great promise for use with other cancer and health populations. PMID- 15789945 TI - Proxy completion of EQ-5D in patients with dementia. AB - There are measurement difficulties associated with the assessment of health related quality of life (HRQL) in older people with dementia. The use of proxies is a commonly employed approach to overcome such problems. The research reported in this paper sought to identify, specifically for the EuroQol EQ-5D HRQL instrument, whether construct validity is greater for 'family caregivers' or 'clinicians' as two alternative sources of proxy information for patients with a diagnosis of dementia. This involved the exploration of the strength of the associations between clinical measures of illness severity and EQ-5D data. The data appear to reveal a pattern suggesting that the viewpoint of the proxy (i.e., clinician or family caregiver) is important. The findings suggest that the data provided by clinicians (when compared to data from carers) had higher construct validity for the more observable dimensions of the EQ-5D instrument (i.e., 'mobility' and 'self-care'). Conversely, the data from family carers had higher construct validity for the less observable dimensions (i.e., 'usual activities' and 'anxiety/depression'). Previous research on proxy provision of HRQL data has tended to focus on trying to identify a single proxy. The results of this study suggest that using carefully matched sets of measures and assessment perspectives may produce more valid EQ-5D health state descriptions. PMID- 15789944 TI - Validation of the EQ-5D quality of life instrument in patients after myocardial infarction. AB - BACKGROUND: We assessed cross-sectional validity of EQ-5D after myocardial infarction (MI). METHODS: We compared EQ-5D, SF-36, quality of life After MI (QLMI), and Canadian Cardiovascular Society Anginal Classification (CCSG) scores. Correlation and regression techniques were used to assess convergent validity. SF 36 and alternate Rand-36 scoring were compared. CCSG class was used to evaluate discriminative validity and clinical difference in health state scores. RESULTS: Of 99 patients: mean age 64; median 176.5 days post-MI; 80% had one MI; 74% were CCSG I. 1/3 to 1/2 reported mobility, self-care, pain, and emotional difficulties on EQ-5D. Median health state was 0.73. EQ-5D and SF-36 (or Rand-36) strongly correlate in overall health (0.75), emotional health (0.75), pain (0.68), and activity/functional (0.5-0.63). EQ-5D and QLMI strongly correlate in activities/self esteem (0.56), emotional health (0.64), anxiety/ depression- restriction (0.53), and overall health (0.5-0.57). EQ-5D self-care correlates weakly with all domains. Domain scores from each general instrument contributed to each other's overall health score (adjusted R2 0.61-0.69) and to disease specific score (0.45 adjusted R2). EQ-5D discriminates among CCSG classes (p < 0.000). Physicians detected a 0.16 difference in health state scores. CONCLUSION: The EQ-5D provides valid general HrQOL measurement post-MI. PMID- 15789946 TI - Variation in Chinese population health related quality of life: results from a EuroQol study in Beijing, China. AB - The purpose of this study is to measure Chinese population health related quality of life (HRQoL) using European quality of life (EQ-5D) instrument, to examine the validity of EQ-5D in measuring Chinese population HRQoL, to explore the relationships between EQ-5D and other health determinants, and to display the similarities and differences of HRQoL between the Chinese population and the populations of other countries. The data used in this study includes 2994 respondents whose age are 12 years and older, which is from the 2000 Beijing Household Health Survey. Univariate and bivariate analyses have been used to examine the level of HRQoL and the relationships between HRQoL and other variables. Multi-variate analyses have been used to explore the relationships between the EQ-5D Visual Analogue Scale (VAS) and the EQ-5D five dimension indicators. There are four principal findings from this study. First, the EQ-5D instrument is a valid measure for Chinese HRQoL, but with a significant ceiling effect. Second, Pain/ Discomfort and Anxiety/Depression are the major Chinese HRQoL problems and the extents of these problems differ in subgroup populations. Third, typically mean scores are lower for older age group; this is observed at lower ages in the Chinese population than in populations from developed countries. Fourth, Chinese HRQoL has strong association relationship with population socio-economic status (SES), which might imply that issues brought on by the rapid economic transition have both positive and negative impacts on Chinese HRQoL. PMID- 15789947 TI - Validation of the FACT-BRM with interferon-alpha treated melanoma patients. AB - The somatic, neurocognitive, and psychiatric side effects of biological response modifiers (BRMs) have been documented in specific patient samples. Although these side effects likely have a predictable impact on patients quality of life (QOL), no instrument currently measures the cumulative effect of the various complaints patients' report. The current study investigated the reliability and validity of the Functional Assessment of Cancer Treatment-Biological Response Modifier (FACT BRM) scale for measuring QOL in a sample of melanoma patients receiving interferon. Measures of distress, depression, and fatigue were also obtained using standardized, well-validated instruments. Results indicate increased symptom burden, depression, and fatigue, and decreased quality of life over 4 months of IFN therapy. The FACT-BRM demonstrated good psychometrics and sensitivity to change, and thus appears to be a good instrument for measuring QOL in patients receiving BRMs. PMID- 15789948 TI - Comparison of WHOQOL-bREF and SF-36 in patients with HIV infection. AB - The purpose of the study was to evaluate the reliability and validity of the two generic instruments, the WHOQOL and the SF-36, for assessing health-related quality of life in 224 patients with HIV infection. The internal consistency ranged from 0.75 to 0.86 across the WHOQOL-BREF domains and from 0.72 to 0.93 across the SF-36 scales. The scores of all WHOQOL-BREF domains and SF-36 scales correlated positively with the measure of happiness, Sat-HRQOL and self-perceived health status, and correlated negatively with the number and intensity of symptoms. Patients with higher CD4 cell counts scored significantly higher on G4 (general health), three WHOQOL-BREF domains, seven SF-36 scales, and PCS (physical component summary). Patients with fewer symptoms and with less intensity of symptoms had significantly higher scores on all four domains of WHOQOL-BREF, eight scales, PCS, and MCS (mental component summary) of the SF-36 scale. The correlations between the physical, psychological, and social domains of the WHOQOL-BREF and PF (physical functioning), MH (mental health), and SF (social functioning) of the SF-36 were 0.51, 0.75, and 0.54, respectively. There is also good correlation between PCS of the SF-36 and the physical domain of the WHOQOL-BREF (r = 0.48), and between MCS and all four domains of the WHOQOL-BREF (r range = 0.60-0.75). The WHOQOL-BREF domains showed fewer floor or ceiling effect than the SF-36 scales. We concluded that both the WHOQOL-BREF and the SF 36 are reliable and valid health related quality-of-life instruments in patients with HIV infection. PMID- 15789949 TI - Content validity, construct validity, and reliability of the WHOQOL-Bref in a population of Dutch adult psychiatric outpatients. AB - In this study, the psychometric properties of a quality of life scale, the WHOQOL Bref, were examined in a population of 533 Dutch adult psychiatric outpatients. Participants underwent two semistructured interviews in order to obtain Axis-I and II diagnoses, according to DSM-IV. Besides the WHOQOL-Bref they also completed questionnaires for measuring psychopathological symptoms (SCL-90) and perceived social support (PSSS). Scores on 25 of the 26 questions of the WHOQOL Bref had a good distribution. Similar to previous findings, exploratory factor analysis revealed a four-factor structure. A priori expected associations were found between the domains of the WHOQOL-Bref, on the one hand, and dimensions of the SCL-90 and the PSSS-score, on the other hand, indicating good construct validity. The internal consistency of the four domains of the WHOQOL-Bref ranged from 0.66 to 0.80. Domain scores of the WHOQOL-Bref correlated around 0.92 with the WHOQOL-100 domain scores. Relatively low correlations were found between demographic characteristics (age and sex) and WHOQOL-Bref domain scores. It is concluded that the content validity, construct validity, and the reliability of the WHOQOL-Bref in a population of adult Dutch psychiatric outpatients are good. The WHOQOL-Bref, therefore, is an adequate measure for assessing quality of life at the domain level in a population of adult psychiatric outpatients. PMID- 15789950 TI - Subjective well-being and community integration among clients of assertive community treatment. AB - Three aspects of community integration (i.e., physical, social, and psychological integration) were examined in relation to subjective well-being (SWB) in a sample of 92 persons with psychiatric disabilities receiving services from assertive community treatment (ACT) teams. Although two of the integration variables, namely physical and psychological integration, were related to SWB at the bivariate level, these relationships were not maintained following sequential multiple regression analysis. Findings support previously established relationships between SWB and psychiatric symptoms and between SWB and self esteem. Results also emphasize the importance of client perceptions of social support that may be attributable, in part, to ACT services. PMID- 15789951 TI - Assessing quality of life in eating disorder patients. AB - OBJECTIVE: To examine quality of life among subgroups of eating disorder patients. METHOD: Self-report questionnaires which included two quality of life measures were completed by 87 individuals referred for treatment to the Australian Capital Territory Eating Disorders Day Program. Health-related quality of life, as measured by the Medical Outcomes Study 12-item Short Form Mental Component Summary scale, and subjective quality of life, as measured by subscales of the World Health Organization Brief Quality of Life Assessment Scale (WHOQOL BREF), were compared among individuals who received the diagnosis of anorexia nervosa purging subtype (n = 15), anorexia nervosa restricting subtype (n = 19), bulimia nervosa (n = 40) and binge eating disorder (n = 10), and among a general population sample of young adult women employed as a control group (n = 495). RESULTS: Eating disorder patients, when considered together, showed marked impairment in both health-related and subjective quality of life relative to normal control subjects. However, in both domains, restricting anorexia nervosa patients reported significantly better quality of life than other patient groups, after controlling for levels of general psychological distress. Scores on the Social Relationships subscale of the WHOQOL-BREF among individuals in this subgroup were similar to those of normal control subjects. CONCLUSIONS: Reliance on any one instrument is likely to be misleading in assessing the quality of life of eating disorder patients. Careful consideration needs to be given to the assessment of restricting anorexia nervosa patients in particular. PMID- 15789952 TI - Psychosocial factors and health-related quality of life in hemodialysis patients. AB - Several sociodemographic and clinical variables are known to influence the health related quality of life (HRQOL) of patients with kidney disease, yet the relationship between psychological factors and the HRQOL measured by the Kidney Disease Quality of Life Short-Form (KDQOL-SF) is incompletely understood. The objective of this study was to examine the relationship between psychosocial status (depressive symptoms, trait anxiety, and social support) and KDQOL-SF scales in hemodialysis (HD) patients by controlling the effects of sociodemographic and clinical variables. The HRQOL of 194 patients from 43 dialysis centers in Spain was assessed by completing the KDQOL-SF, and evaluating depressive Symptoms (Cognitive Depression Index), trait anxiety (Trait Anxiety Inventory) and degree of social support (Scale of Perceived Social Support). We also recorded several sociodemographic and clinical variables. Two regression models were estimated for each of the 19 scales in the KDQOL-SF. In the first model, we only included sociodemographic and clinical-factors, while the second model also took into consideration psychosocial variables. These last factors (trait anxiety and depressive symptoms, not social support) were found to increase the proportion of explained variability, with highest standardized regression coefficients observed for most KDQOL-SF scales. Depressive symptoms were related to a poor HRQOL when there was a strong physical component, while trait anxiety was mainly related to emotional upset and social relationships. We were able to conclude that trait anxiety and depressive symptoms are strongly associated with the HRQOL assessed by the KDQOL-SF in HD patients. The effects of these factors should therefore be considered when evaluating the quality of life of this type of patient. PMID- 15789953 TI - Factorial design for the analysis of patient's quality of life in asthma. AB - Chronic diseases like asthma have a great impact on the everyday life of patients. Evaluation of a patient's quality of life informs us about how the patient is feeling and whether or not he or she is able to function physically, socially and emotionally. The objectives of this study were to assess the quality of life of asthmatic patients and to study the influence of age and disease severity, as independent variables, on the patient's quality of life. A 2-factor, 3-level face-centred central composite design was applied to construct a second order polynomial model describing the effects of age and the severity on the quality of life. The second-order polynomial model was successfully applied to describe the effect of age and asthma severity with good correlation on the quality of life measured by either generic (Visual analogue scale) or disease specific (St George's Respiratory Questionnaire) instruments in the examined subjects. Our findings suggest quality of life may be used as a non-invasive patient-centred monitoring system as a guide for disease management if measured regularly. PMID- 15789954 TI - Effectiveness of dental services in facilitating recovery from oral disadvantage. AB - OBJECTIVE: To determine the effectiveness of dental health care services in facilitating recovery from quality of life decrements in symptom-specific subgroups with the most prevalent chronic infectious diseases (periodontal disease and dental caries), and a 'stained teeth' subgroup. METHODS: Data were taken from the prospective longitudinal Florida Dental Care Study of 873 individuals 45+ years old. Logistic regression modeling quantified associations between recovery from oral health-related quality of life decrements ('recovery') and dental services. RESULTS: Adjusting for age, race, gender, income, approach to dental care, and signs/symptoms, any dental visit (odds ratio, OR: 4.0; 95% confidence interval, CI: 2.3, 6.9), corrective treatment (OR: 3.8; 95% CI: 1.6, 8.7), denture visit (OR:4.8; 95% CI: 1.1, 21.9), or extraction (OR: 6.2; 95% CI: 2.2, 17.4) were positively associated with recovery. Upon conditioning the analyses on specific symptoms, point estimates increased substantially for most service types, and dental cleaning was associated with recovery for the stained teeth subgroup (OR: 10.9; 95% CI: 1.2, 99.4). CONCLUSION: Dental care was highly effective in treating quality of life decrements. Treatment effectiveness increased substantially when analyses were restricted to symptom-specific subgroups similar to selection criteria of randomized clinical trials (RCTs). Restricted cohort analyses can be applied to many other health outcomes for which RCTs are not feasible or ethical. PMID- 15789955 TI - Responsiveness of generic health-related quality of life measures in stroke. AB - OBJECTIVE: To compare five preference-based generic measures of health-related quality of life (HRQOL) in terms of change scores, correlations among change scores, responsiveness, and quality adjusted life-years (QALYs) gained. DESIGN: Observational longitudinal cohort study where clinical measures and self-assessed HRQOL measures were administered to stroke patients at baseline and at 6 months. Patients were categorized as 'stable', 'some improvement' and 'large improvement' using the Barthel Index, Modified Rankin Scale (MRS), and Center for Epidemiologic Studies Depression Scale (CES-D). For each group, paired t-tests and variants of effect size were used to compare the responsiveness of preference based HRQOL summary scores, including the EQ-5D VAS and index-based score, SF-6D, and Health Utilities Index (HUI) Mark 2 (HUI2) and Mark 3 (HUI3) overall utility scores. RESULTS: Ninety-eight of 124 (79%) patients completed the 6-month follow up. Change scores of the EQ-Index, HUI2, and HUI3 were strongly correlated with changes in the Barthel Index and MRS, while the EQ-5D VAS had higher correlation with CES-D change scores than the other measures. The SF-6D, HUI3, and EQ-Index were generally more responsive than the HUI2 and EQ-5D Visual analogue scale (EQ VAS). QALY estimates based on the EQ-5D index and HUI3 were twice as large as estimates based on the SF-6D and HUI2. CONCLUSIONS: The results of this study may assist in informing the selection of a preference-based generic HRQOL measure, although choice will also depend on study goals and context. We would caution against the generalization of the study results on responsiveness to conditions when more subtle change is expected. PMID- 15789956 TI - Subjective versus interviewer assessment of global quality of life among persons with schizophrenia living in the community: a Nordic multicentre study. AB - BACKGROUND: Few studies have investigated differences between subjective and externally assessed quality of life in individuals with a severe mental illness. In a sample of 387 patients with schizophrenia living in the community the present study investigated the association between subjective and interviewer rated quality of life, clinical and sociodemographic factors related to the two assessments, and if discrepancies in the assessments were related to any clinical or social features of the patients. METHOD: The study was a Nordic multicentre study with a cross-sectional design. Instruments used were the Lancashire Quality of Life Profile, the Brief Psychiatric Rating Scale, the Interview Schedule for Social Interaction, Camberwell Assessment of Needs and General Assessment of Functioning. RESULTS: The correlation between subjective and interviewer-rated quality of life was moderate (ICC = 0.33). More severe affective symptoms, fewer emotional relations and a lower monthly income were related to poorer subjectively rated quality of life but in a stepwise multiple regression analysis accounted for only 14.1% of the variance. Poorer interviewer-rated quality of life was mainly related to a more severe psychopathology but also to a lower monthly income, fewer emotional relations and not being employed. Together these factors accounted for 45.5% of the variance. A greater discrepancy between the subjective and the interviewer rating was found in patients with less affective symptoms, unemployment, and a better social network. CONCLUSION: Only a moderate correlation between subjective and interviewer-assessed global quality of life was found, implying that the sources of assessment differed, as was also shown in subsequent regression models. It is concluded that both perspectives on the patient's quality of life may be valuable for treatment planning, especially in cases where differences in quality of life assessment related to the patient's psychopathology may be expected. PMID- 15789957 TI - International development of the Parents' Index of Quality of Life in Atopic Dermatitis (PIQoL-AD). AB - OBJECTIVE: The international development of the Parents' Index of Quality of Life in Atopic Dermatitis (PIQoL-AD), a quality of life (QoL) instrument specific to parents of children with atopic dermatitis (AD) is described. METHOD: The instrument was developed simultaneously in several countries. Its content was derived from 65 qualitative interviews with parents in the UK, Netherlands and Italy. The measure was then produced for the UK, Netherlands, Italy, Germany, France, US and Spain. Field-test interviews were conducted with approximately 20 patients in each country to assess face and content validity. A two time-point survey was conducted with between 45 and 328 parents in each country to finalise the instrument through application of the Rasch model and to evaluate the psychometric properties of the final instrument. RESULTS: Application of the Rasch model to the survey data identified the final 28-item version. All language versions had good item fit, test-retest reliability (above 0.85), internal consistency and promising validity. CONCLUSIONS: The PIQoL-AD is a valuable instrument for inclusion in clinical trials and routine clinical practice. It provides distinct and complementary information to that of existing dermatology specific measures and has been shown to be responsive to changes in QoL in clinical trials. PMID- 15789958 TI - Development, reliability and validity of a new measure of overall health for pre school children. AB - BACKGROUND: Few comprehensive systems are available for assessing and reporting the overall health of preschool children. OBJECTIVES: (i) To develop a multi dimension health status classification system (HSCS) to describe pre-school (PS) children 2.5-5 years of age; (ii) to report reliability and validity of the newly developed measure. DESIGN: Existing systems (Health Utilities Index, Mark 2 and 3) were adapted for application to a pre-school population. The new system was tested for acceptability, validity and reliability. PARTICIPANTS: Three cohorts of children and their parents from Canada and Australia were utilized: Cohort 1 (MAC)-101 3-years old very low birthweight (VLBW, <1500 g) and 50 same age term children from Canada; Cohort 2 (AUS)-150 VLBW 3-years old from Australia; Cohort 3 (OMG)-222 3-years old with cerebral palsy (CP) from Ontario. METHODS: Parental intra-rater reliability was evaluated by completion of the HSCS-PS Parent questionnaire (MAC) at the clinic visit and again 14 days later. Health professionals (MAC) completed the HSCS-PS Clinician questionnaire. Percent agreement and Kappa values were used to assess parent-clinician agreement. Concurrent validity was tested in two populations of VLBW children (MAC and AUS) and a reference group of term children (MAC) by exploring the relationships between dimensions of the HSCS-PS and well-recognized norm-referenced measures: the Bayley Scales of Infant Development (BSID-II), the Vineland Adaptive Behavior Scales (VABS) and the Stanford-Binet (SB). Construct validity was tested by comparing ratings on both the HSCS-PS and the Gross Motor Function classification system (GMFCS) using a population of pre-school children with CP. Analyses were done using chi2, ANOVA and correlations with tau-b statistic. RESULTS: The HSCS PS has 12 dimensions and 3-5 levels per dimension. Response rate for parental intra-rater reliability was 95%, with percent agreement ranging between 86 and 100%. Kappa values for various dimensions ranged from 0.38 to 1.00. Inter-rater reliability between parents and clinicians showed agreement ranging from 72 to 100%. Kappa values ranged from 0.30 to 1.00. CONCURRENT VALIDITY: There was a statistically significant gradient between HSCS-PS Mobility levels and motor scale scores of the BSID-II and VABS. A significant gradient also occurred when comparing HSCS-PS cognition levels to psychometric scores on the BSID-II and SB, as well as HSCS-PS self-care levels compared to VABS Daily Living scores. DISCRIMINATIVE AND CONSTRUCT VALIDITY: Birthweight category was shown to be a significant determinant of proportion of children with multiple HSCS-PS dimensions affected. In addition, HSCS-PS dimension levels were congruent with GMFCS levels where expected: mobility had excellent correlation; self-care, dexterity, speech and cognitive dimensions had moderate correlations. CONCLUSIONS: The HSCS-PS is readily accepted, quick to complete, widely applicable and provides a multi-dimensional description of health status. Preliminary assessments of reliability and validity are promising. The HSCS-PS can discriminate across populations by birthweight and shows strong relationships with standardized psychometric measures in comparable domains. It can pro- vide a summary profile of functional limitations in various populations of pre-school children in a consistent manner across programs and in different settings. PMID- 15789959 TI - Reliability and construct validity of the SF-36 in Turkish cancer patients. AB - In this study, we assessed the reliability and construct validity of the SF-36, Turkish version on 419 cancer patients. Cronbach's alpha coefficients surpassed the 0.70 criterions for all subscales indicating good internal consistency. Results of the test-retest method showed that the stability coefficients for the eight subscales of the SF-36 ranged between 0.81 and 0.94. Principal components factor analysis with varimax rotation confirmed the presence of seven factors in the SF-36: physical functioning, role limitations due to physical and emotional problems, mental health, general health perception, bodily pain, social functioning, and vitality. In conclusion, the Turkish version of the SF-36 is a suitable instrument that could be employed in cancer research in Turkey. PMID- 15789960 TI - Responsiveness, longitudinal- and cross-sectional construct validity of the Pediatric Asthma Quality of Life Questionnaire (PAQLQ) in Dutch children with asthma. AB - OBJECTIVE: Health-related quality of life is an important measure in evaluations of the management of childhood asthma. In this study, we assessed psychometric properties, responsiveness, and longitudinal and cross-sectional construct validity of the Dutch version of the 23-item Pediatric Asthma Quality of Life Questionnaire (PAQLQ). METHODS: The study group consisted of 238 6-18-year olds with asthma, with complete respiratory symptom diaries in the course of one winter season; each child had one (or more) PAQLQ measurement(s) concerning one (or more) week(s) with relatively many symptoms (n = 238). Each child also had one PAQLQ measurement concerning another week with relatively few symptoms (n = 238). The PAQLQ scores of the 238 children for a week with few symptoms (the symptom diary scores remained below a predefined level everyday) were compared with their PAQLQ scores for another week with many symptoms (on day 1 of that week, symptom diary scores had been above the predefined level). Additionally, in a subgroup of the study group that had experienced two or more 'weeks with many symptoms' (n = 101), we compared the PAQLQ-scores for two different weeks with many symptoms of these children. RESULTS: Only the domain Emotions showed a ceiling effect (>25% had the maximum score). All Cronbach's alpha's of the PAQLQ total score and domains were >0.70, except for Activities (alpha = 0.54). Mean PAQLQ-scores were significantly different (p < 0.01; n = 238) between one week with few symptoms and another week with many symptoms. Contrary, in the subgroup of children with PAQLQ-measurements regarding more than one week with many symptoms (n = 101), mean PAQLQ-scores did not differ significantly (p > or = 0.05) between 1 week with many symptoms and another week with many symptoms. These results indicate responsiveness. (Changes in) lower respiratory tract symptoms, indicative of asthma severity, correlated better with (changes in) PAQLQ scores than (changes in) upper respiratory tract symptoms, which supports the longitudinal and cross-sectional construct validity. CONCLUSION: The assessed properties of the PAQLQ linguistic validation into Dutch were similar to those originally established for the PAQLQ in Canada. This study showed that the Dutch PAQLQ has adequate psychometric properties, excellent responsiveness, and that the longitudinal and cross-sectional construct validity is supported. PMID- 15789962 TI - Dosage clarification. PMID- 15789961 TI - Validity and reliability of the Chinese (Singapore) version of the Parkinson's Disease Questionnaire (PDQ-39). AB - OBJECTIVE: The purpose of the study was to validate the Chinese (Singapore) version of the Parkinson's Disease Questionnaire (PDQ-39CSV) and its briefer version (PDQ-8CSV). METHODS: A convenience sample of Chinese-speaking Singaporeans with Parkinson's disease (PD) (n = 63) completed a questionnaire containing the PDQ-39CSV and the Chinese (Singapore) EQ-5D. A subgroup also participated in a retest and/or a focus group discussion. A priori hypotheses were tested by examining correlations between PDQ-39CSV, PDQ-8CSV and EQ-5D scores and using principal component factor analysis. Reliability was assessed using Cronbach's alpha and intra-class correlation coefficients (ICC). RESULTS: Thirty-two PDQ-39CSV items correlated satisfactorily with their hypothesized dimensions (Spearman's p > or = 0.4). Factor analysis yielded a component on which all 8 PDQ-39CSV dimensions were substantially loaded (loading range: 0.53 0.89). As hypothesized, the PDQ-39CSV and PDQ-8CSV summary indices were highly correlated (Pearson's r:0.95, ICC:0.94); correlations between related PDQ and EQ 5D scores were generally strong (Spearman's p: 0.38-0.76, p < 0.001 for all). Cronbach's alpha values ranged from 0.64 to 0.90 and ICC values from 0.66 to 0.86. CONCLUSION: This study provides preliminary evidence supporting validity and reliability of both the PDQ-39CSV and its briefer version. PMID- 15789963 TI - The importance of agrinursing. PMID- 15789964 TI - Surgical smoke--is there a fire? PMID- 15789965 TI - Worksite influenza immunization programs. Insight into the implementation and cost-benefit. AB - The most important factors in cost-benefit analyses of worksite influenza immunization programs are vaccine cost and administration, vaccine efficacy, and the assumptions used to calculate lost productivity from worker absenteeism or decreased effectiveness. Based on several models published in the literature, estimates of savings per healthy working adult employee could be anticipated to range between dollars 15 and dollars 50. Variations of this net savings are primarily based on assumptions of productivity estimates of the employee. Efficient large worksite influenza immunization programs are feasible and worthy of employer consideration for economic and employee satisfaction reasons. PMID- 15789966 TI - Impact of converting to powder-free gloves. Decreasing the symptoms of latex exposure in operating room personnel. AB - This study examined health care worker satisfaction with the use of non-powdered natural rubber latex (NRL) surgical gloves to determine the impact of non powdered NRL gloves on the NRL sensitization of operating room personnel. The study used a 1-year longitudinal design to obtain recall information from employees about their NRL exposure. Additionally, a survey was completed by participants related to their satisfaction with non-powdered NRL gloves. Informed consent was obtained from 103 employees. After conversion to an operating room using non-powdered NRL, there was a significant decrease in reported symptoms with NRL exposure (42% pre- and 29% post-conversion, Fisher's exact, two-tailed, p = .0001). This study demonstrated that the conversion to non-powdered low protein NRL gloves resulted in decreased symptoms because of NRL exposure. PMID- 15789967 TI - Needlestick injuries in the United States. Epidemiologic, economic, and quality of life issues. AB - Best evidence from prospective studies with aggressive monitoring suggests that the incidence of needlestick injuries is significantly higher than reported through passive surveillance, ranging from 14 to 839 needlestick injuries per 1,000 health care workers per year. The economic cost of managing these injuries is substantial, ranging from dollars 51 to dollars 3,766 (2002 U.S. dollars). This amount excludes the cost of treating the long-term complications of needlestick injuries, such as HIV and hepatitis B and C infections, each of which can cost several hundreds of thousands of dollars to manage. In addition, health care workers experience significant fear, anxiety, and emotional distress following a needlestick injury, sometimes resulting in occupational and behavior changes. Despite the availability of engineered injury prevention devices, the implementation of these new technologies has been mixed in part because of the perception that these devices are costly and cost ineffective. However, widespread use of safety devices might be more easily justified on economic grounds when the full clinical and economic benefits of these new technologies are considered, especially within the context of injury prevention. PMID- 15789968 TI - Frequent flyer business travelers. The role of the occupational health nurse. AB - When managing frequent flyer business travelers, occupational health nurses focus on health promotion and health protection goals. The three types of prevention (i.e., primary, secondary, tertiary) follow a timeline beginning with complete prevention, and proceeding through and ending with management of a disease process. Occupational health nurses design and implement practice strategies based on this progression. Travel health nursing is rapidly expanding as the number of travelers, immunizations, and modes of transportation increase. Physicians focus on disease, industrial hygienists focus on hazard exposure, and safety professionals address occupational issues related to illnesses and injuries. Occupational health nurses are the professionals who focus on all three areas, in addition to health promotion and health protection. Frequent flyer business travelers have specific and complex needs that occupational health nurses are in a unique position to address. PMID- 15789969 TI - Daily fiber intake--how much is enough? PMID- 15789970 TI - What determines the normal water content of a living cell? AB - Most living cells contain a large amount of water. To improve our understanding of this fundamental phenomenon of cell physiology, five theories are critically examined in the light of three sets of relevant experimental findings. These findings are: (1) the diversity and specificity of the percentage water content to tissue type; (2) the limitation imposed by the Law of the Conservation of Energy on postulating membrane pumps and (3) the non-extractability of cell water from the open ends of muscle cells whose membrane covering has been surgically removed. Two of the five theories examined are called respectively the accidental theory (Theory I) and the direct water pump-leak theory (Theory III); both are introduced for the first time here as working hypotheses. Three others theories examined were published; they comprise the Donnan membrane equilibrium theory (Theory II), the indirect pump-leak (Theory IV) and the polarized-oriented multilayer (PM) theory of cell water (Theory V.) The PM (Theory V) alone is in harmony with, and supported by all three sets of the experimental findings. The remaining theories are shown to be non-applicable to cell water by at least two of the findings PMID- 15789972 TI - Evaluation of a simple carrier molecule to enhance drug penetration of dermal layers by utilizing multivariate methods, structure property correlations, and continuous system modeling. AB - Nicotinic acid is shown to be comparable to dihydropyridine in its capacity to facilitate penetration of an attached antibacterial drug through dermal layers. Antibacterial drugs examined with nicotinic acid or dihydropyridine carriers were beta-lactam antibiotics: methicillin, oxacillin, benzylpenicillin, penicillin F, penicillin dihydro F, propicillin, carbenicillin, penicillin K, penicillin X, and ampicillin. An oxymethyl (-O-CH2-) group is inserted as the linker between the antibiotic and the carrier group. Structure Property Correlations and multivariate methods such as regression analysis, cluster analysis, principal component analysis, discriminate analysis, self-organizing tree algorithm, and factor analysis clearly showed that nicotinic acid performs as an effective carrier drug and is comparable to dihydropyridine. The skin penetration constant Kp was calculated for all 10 antibiotics having either dihydropyridine or nicotinic acid as carrier, and was found to have a mean of 5.13E-05 cm/hour and 1.83E-05 cm/hour, respectively. The standard deviation for each group showed the numerical values overlap as did the 90% confidence interval for each group. A hierarchical tree organization of skin shows the overlapped dermal layers as they exist in normal skin and for the model utilized in this work. A Deming-Regression analysis also shows the nicotinic acid and dihydropyridine structures to have similar and correlated water solubility. Plotting Kp of the dihydropyridine structures as independent variable versus Kp of the nicotinic acid structures show good correlation (Pearson correlation r = 0.6606) and no significant departure from linearity. Connected box plots showed the majority of Kp values for each group of modified antibiotics to exist in a tight cluster. Polar graph of the Log Kow values showed the two groups of modified antibiotics to be correlated and numerically adjacent in trend. ChemSketch property calculations and modeling demonstrates the affects of structural oxygens, nitrogens, carbonyl groups, amide groups, and aromatic rings that are important in understanding the pervasiveness through dermal layers. Continuous model analysis by acslXtreme is utilized and demonstrates three models of the dispersion of drugs through dermal layers based on diffusivity constant (D), Log Kp from Log Kow and formula weight, and Kp as a function of time. PMID- 15789971 TI - 17beta-estradiol suppresses ROS-induced apoptosis of CHO cells through inhibition of lipid peroxidation-coupled membrane permeability transition. AB - Oxidative stress-induced apoptotic cell death has been implicated to play a critical role in the mechanism of corpus luteum regression and follicular atresia. Recent studies suggests that reactive oxygen species (ROS) might play important roles in the regulation of luteal function. The present work describes the inhibitory effect of 17beta-estradiol (E2) on ROS-induced mitochondrial membrane permeability transition (MPT) and apoptosis of Chinese hamster ovary (CHO) cells. ROS generated by Fe2+ and H2O2 induced mitochondrial lipid peroxidation, depolarization, activation of caspase-3 and DNA fragmentation in CHO cells by some E2-inhibitable mechanism. E2 suppressed the Fe2+/H2O2-induced lipid peroxidation and MPT of isolated mitochondria that was characterized by cyclosporin A-inhibitable swelling, depolarization and cytochrome c release. Furthermore, E2 scavenged the xanthine oxidase generated ROS. These results suggests that Fe2+/H2O2 induced MPT and apoptosis of CHO cells by a mechanism that could be suppressed by antioxidant properties of E2. PMID- 15789973 TI - The thermoelectric properties of hornet cuticle: correlation with measuring body sites and activity status. AB - Our study focused on the thermoelectric properties of hornet cuticle at different body compartments and under varying states of awakeness. We also measured the temperature alteration patterns in various body parts of the hornet. Electric voltage and current were dependent on: a) the state of wakefulness; b) the part of the body. The current was lowest in dead hornet cuticle, somewhat higher in narcotized hornet cuticle, considerably higher in the cuticle of hornets awakening from anesthesia and highest in fully awake hornets. Voltage values were of the same order for dead and narcotized hornets, but considerably higher in unanesthetized awake hornets and highest in the cuticle of hornets awakening from anesthesia. At optimal temperature (29 degrees C) the hornet body temperature was higher on the abdominal cuticle than on other body parts. At an ambient temperature of 20 degrees C, the highest temperatures were recorded on the head and thorax, and the lowest on the abdomen. Body temperatures of live hornets were higher than the cooler ambient temperature outside the nest at night. The results suggest that the hornets possess an intrinsic biological heat pump mechanism, which can be used to achieve active thermoregulation. PMID- 15789974 TI - On the modulation effect of pulsing and static magnetic fields and mechanical vibrations on barley seed hydration. AB - The changes of wet and dry weights of barley seed in different periods of swelling were studied in seeds treated with Extremely Low Frequency Electromagnetic Fields (ELF EMF), Static Magnetic Fields (SMF) and Mechanical Vibrations (MV) in cold (4 degrees C) and warm (20 degrees C) distilled water as well as in seeds non-treated (control). The metabolic dependent seed hydration, dry weight loss and water binding in seed were modulated by preliminary EMF, SMF and MV-induced treatment of distilled water. The specific electrical conductivity (SEC) of control and treated distilled water was measured before the seed incubation. Frequency and intensity "windows" (i.e. range of frequency or intensity) for the effect of EMF, MV and SMF (correspondingly) on seed hydration, solubility and water binding in seed were studied. These "windows" were different in various phases of seed swelling. It is suggested that water structure modification is the result of valence angle changes (SMF and EMF) and dipole molecules vibration (EMF and MV) has different effects on the process of hydration, solubility and water binding in seed. These results are important from the point of understanding the mechanisms of the biological effect of EMF, as well as from the point of agriculture. PMID- 15789975 TI - Biology of the oral mucosa. PMID- 15789976 TI - Driving toothbrush innovation through a cross-functional development team. AB - A cross-functional team was assembled to take on the project of developing a toothbrush that could deliver a "whole mouth clean." As such, this brush needed to not only demonstrate superior plaque removal from teeth, it also needed to provide a means for safe cleaning of soft oral tissues. During use, the brush needed to be comfortable both in hand and in mouth, and it required contemporary esthetics to help drive consumer acceptance. This article will take the reader through the full development cycle of the new toothbrush, identifying the proposed benefit of each brush feature along the way. Where appropriate, care was taken to identify the specific insights leading to the development of several new features not previously seen in commercial toothbrushes. This article also will attempt to demonstrate the value of forming new product development teams with representatives of multiple functional areas, including marketing, product development, advanced technology, clinical, engineering, and manufacturing. PMID- 15789977 TI - Clinical efficacy of Colgate 360 degrees and three commercially available toothbrushes on the removal of desquamated epithelial cells. AB - A clinical study was done to evaluate the performance of four toothbrushes on the removal of desquamated epithelial cells after brushing according to the manufacturers' instructions for use. This randomized, crossover-design clinical study compared a new manual toothbrush (Colgate 360 degrees) to two commercially available manual toothbrushes (Oral-B CrossAction and Oral-B Indicator) and a commercially available battery-powered toothbrush (Crest SpinBrush PRO). Adult men and women subjects reported to the clinical facility after a 1-week "washout" period of brushing with a regular fluoride dentifrice and a soft-bristled toothbrush. Participants reported having refrained from oral hygiene procedures, eating, and drinking that morning. After providing a baseline rinse sample, subjects brushed their teeth for 1 minute with their assigned toothbrush and a commercially available fluoride toothpaste, then returned 30 minutes later to provide postuse rinse samples. Subjects refrained from dental hygiene, eating, and drinking during the 30-minute evaluation period. To provide the samples, subjects rinsed with 10 mL of sterile phosphate-buffered saline solution for 10 seconds. Each collected sample was centrifuged, resuspended, and run in a colorimetric assay to determine the level of desquamated epithelial cells found in the rinse as measured by the absorbance at 570 nm. Twenty adults completed the study. At baseline, the mean levels of desquamated epithelial cells for the 4 treatments were 0.70+/-0.27, 0.63+/-0.20, 0.69+/-0.30, and 0.62+/-0.31 for the Colgate 360 degrees, Oral-B Indicator, Crest SpinBrush PRO, and Oral-B CrossAction, respectively. Posttreatment, the mean levels of epithelial cells were 0.19, 0.38, 0.42, and 0.34, respectively. All of the treatments provided a statistically significant reduction compared to their respective baseline. In addition, the Colgate 360 degrees toothbrush was statistically significantly better than the other three toothbrushes in reducing desquamated epithelial cells. Therefore, the results of this randomized, crossover clinical study indicate that the newly designed Colgate 360 degrees manual toothbrush, with a tongue-cleaning implement on the back of the brush head, was statistically significantly more effective than the Oral-B Indicator, Crest SpinBrush PRO, and Oral-B CrossAction toothbrushes in removing desquamated epithelial cells. PMID- 15789978 TI - Clinical comparison of a new manual toothbrush on the level of hydrogen-sulfide forming bacteria on the tongue. AB - The objective of this randomized, crossover study was to compare the effectiveness of a newly designed manual toothbrush (Colgate 360 degrees) to two commercially available manual toothbrushes (Oral-B Indicator and Oral-B CrossAction) and a battery-powered toothbrush (Crest SpinBrush PRO) for their ability to reduce hydrogen-sulfide-forming bacteria on the tongue. After a washout period, subjects arrived at the clinical site for baseline sampling without performing dental hygiene, eating, or drinking. Subjects sampled the left side of their tongue with a cotton swab. Subjects brushed for 1 minute with the assigned test toothbrush and regular fluoride toothpaste. Those using the Colgate 360 degrees toothbrush were instructed to clean their tongue with the implement on the back of the brush head for 10 seconds. After 2 hours, the subjects returned to the clinical site having refrained from dental hygiene, eating, and drinking for posttreatment sampling, this time sampling the right side of their tongue. After a minimum 2-day washout period, subjects repeated the same regimen using the other toothbrushes. Collected tongue samples were dispersed in sterile water, serially diluted in sterile phosphate-buffered saline, and plated in duplicate onto lead acetate agar. When plated on this medium, bacteria that produce hydrogen sulfide appear as dark-pigmented colonies. After 72 hours of incubation, the dark colonies were counted, expressed as log colony-forming units/mL, and reduction from baseline was calculated. Thirty-one adult men and women completed the clinical study. There was no significant difference between baseline hydrogen-sulfide-forming bacteria levels. Posttreatment, the log reduction of bacteria was 0.80, 0.41, 0.33, and 0.44 for the Colgate 360 degrees, Oral-B Indicator, Crest SpinBrush PRO, and Oral-B CrossAction, respectively. Statistical analysis indicated that the Colgate 360 toothbrush was statistically significantly better (P < .05) than the 3 commercial toothbrushes in reducing the levels of hydrogen-sulfide-forming bacteria on the tongue. PMID- 15789979 TI - Clinical comparison of a new manual toothbrush on breath volatile sulfur compounds. AB - The objective of this randomized, crossover study was to compare the effectiveness of a newly designed manual toothbrush (Colgate 360 degrees) to two commercially available manual toothbrushes (Oral-B Indicator and Oral-B CrossAction) and a battery-powered toothbrush (Crest SpinBrush PRO) for their ability to reduce overnight volatile sulfur compounds (VSC) associated with oral malodor. The study followed a four-period crossover design. Following a washout period, prospective subjects arrived at the testing facility without eating, drinking, or performing oral hygiene for baseline evaluation of breath VSC levels. For each phase of the study, subjects were given one of the test tooth brushes and a tube of regular toothpaste to take home, and they were instructed to brush their teeth in their customary manner for 1 minute. When using the Colgate 360 degrees toothbrush, subjects were instructed to clean their tongue with the implement on the back of the brush head for 10 seconds. The following morning, subjects reported to the testing facility, again without performing oral hygiene, eating, or drinking, for the overnight evaluation. After a minimum 2-day washout period, subjects repeated the same regimen using the other toothbrushes. The levels of breath VSC were evaluated instrumentally using a gas chromatograph equipped with a flame photometric detector. Measurements were taken in duplicate and then averaged. The levels of VSC were expressed as parts per billion (ppb) in mouth air. Sixteen men and women completed the study. At baseline, the mean levels of VSC in mouth air for the 4 toothbrushes were 719.8 ppb+/-318.4 ppb, 592.8 ppb+/-264.6 ppb, 673.8 ppb+/-405.9 ppb, and 656.2 ppb+/-310.2 ppb for the Colgate 360 degrees, Oral-B Indicator, Crest SpinBrush PRO, and Oral-B CrossAction, respectively. Overnight, the mean breath VSC levels after using the four toothbrushes were lower than those observed at baseline. The respective mean levels of breath VSC were 266.5 ppb+/-269.9 ppb, 545.2 ppb+/-346.1 ppb, 567 ppb+/ 335.7 ppb, and 554.6 ppb+/-398.4 ppb. Only the Colgate 360 degrees toothbrush provided a statistically significant reduction (P < .05) in breath VSC vs baseline. Additionally, the Colgate 360 degrees toothbrush was statistically significantly better (P < .05) than the three commercial toothbrushes in reducing breath VSC. Therefore, the results of this randomized, crossover clinical study indicate that a newly designed manual toothbrush with a tongue-cleaning implement on the back of the brush head was significantly more effective than three commercially available toothbrushes in reducing morning breath VSC associated with oral malodor. PMID- 15789980 TI - Comparison of the clinical efficacy of a new manual toothbrush on gingivitis reduction and plaque removal. AB - The objective of this controlled, examiner blind, 4-week clinical study was to evaluate and compare the safety and efficacy of a newly designed manual toothbrush, the Colgate 360 degrees toothbrush, to the Oral-B Indicator toothbrush for the control of supragingival plaque and gingivitis. A total of 82 subjects from the northern New Jersey area reported to the clinical facility for a baseline plaque and gingivitis examination after having refrained from all oral hygiene procedures for 12 hours and from eating, drinking, or smoking for 4 hours. The population was comprised of healthy adult men and women 30 to 68 years of age. After the baseline examinations, qualifying subjects were randomized into two groups and assigned to one of the two test toothbrushes. All subjects were instructed to brush their teeth for 1 minute under supervision, after which they were again examined for supragingival plaque. They were then instructed to brush their teeth twice a day for 1 minute with their assigned toothbrush and a commercially available toothpaste (Colgate Cavity Protection Great Regular Flavor Fluoride Toothpaste) for the next 4 weeks. After 4 weeks, subjects returned to the clinical facility for a final gingivitis and plaque examination. Eighty-one subjects complied with the protocol and completed the 4-week clinical study. The results of the study indicated that the new manual toothbrush was statistically significantly effective in reducing gingivitis after 4 weeks and in removing plaque after a single toothbrushing and after 4 weeks of use. Also, the new manual toothbrush exhibited a statistically significant greater reduction in gingivitis and in gingivitis-related bleeding sites after 4 weeks of use as well as statistically significant greater plaque removal after a single toothbrushing and after 4 weeks of use, as compared to the Oral B Indicator toothbrush. This superior plaque-removal performance was found in separate analyses of the whole mouth, at interproximal surfaces, and at the gumline. PMID- 15789981 TI - A clinical comparison of the gingivitis reduction and plaque-removal efficacy of a new manual toothbrush. AB - The objective of this controlled, examiner-blind, 4-week clinical study was to evaluate and compare the safety and efficacy of a newly designed manual toothbrush (Colgate 360 degrees toothbrush) to the Oral-B CrossAction toothbrush for the control of supragingival plaque and gingivitis. A total of 80 subjects from the central New Jersey area reported to the clinical facility for a baseline plaque and gingivitis examination after having refrained from all oral hygiene procedures for 12 hours and from eating, drinking, or smoking for 4 hours. The population was comprised of healthy adult men and women 18 to 67 years of age. After the baseline examinations, qualifying subjects were randomized into two groups and assigned to one of the two test toothbrushes. All subjects were instructed to brush their teeth for 1 minute under supervision, after which they were again examined for supragingival plaque. They were then instructed to brush their teeth twice a day for 1 minute with their assigned toothbrush and a commercially available toothpaste (Colgate Cavity Protection Great Regular Flavor Fluoride Toothpaste) for the next 4 weeks. After 4 weeks of use, subjects returned to the clinical facility for a final gingivitis and plaque examination. Seventy-eight subjects complied with the protocol and completed the 4-week clinical study. The results of the study indicated that the new manual toothbrush was statistically significantly effective in reducing gingivitis after 4 weeks and in removing plaque after a single toothbrushing and after 4 weeks of use. Also, the group using the new manual toothbrush exhibited a statistically significant greater reduction in plaque of up to 40% and no statistically significant difference in gingivitis reduction after 4 weeks of use, as compared to the Oral-B CrossAction toothbrush. PMID- 15789983 TI - Promoting the positive aspects of dentistry. PMID- 15789982 TI - Laboratory investigation of Colgate 360 degrees toothbrush and Oral-B indicator toothbrush for the removal of dental stains. AB - The objective of this in vitro study was to evaluate the stain-removal efficacy of a newly designed manual toothbrush, the Colgate 360 degrees, relative to a commercially available toothbrush, the Oral-B Indicator. A modification of Stookey et al was used to evaluate the stain-removal effects of toothbrushes instead of dentifrice on bovine teeth. A V.8 mechanical cross-brushing machine equipped with the test toothbrushes and adjusted to 500g to enamel surfaces evaluated stain removal using a dentifrice slurry and water after 800 double strokes. The overall results of this laboratory investigation indicate that the Colgate 360 degrees toothbrush is more effective, P < .05, than the commercial Oral-B Indicator toothbrush in removing dental stain and brightening teeth using a standard toothpaste or water. PMID- 15789984 TI - AIDS: the attitudes and experience of final year European dental students. PMID- 15789985 TI - Parents' preference as to whether they would like to accompany their child when receiving dental treatment--results from a national survey. AB - BACKGROUND AND DESIGN: parental accompaniment of children during dental treatment has always been a contentious issue. One of the factors that should be considered is the preference of the parents. The aim of this paper is to report the preference of parents of eight-year-old schoolchildren in Ireland in relation to accompanying their child during dental procedures and is part of the National Survey of Children's Dental Health in Ireland, which was conducted between October 2001 and June 2002. The survey had a cross sectional design and parents of children (average age 8.4 years) selected for dental examination were asked to complete a questionnaire. RESULTS: 3629 completed questionnaires were obtained from parents of eight-year-old children giving a response rate of 68 per cent. Sixty-seven per cent of parents expressed a preference to accompany their child during dental treatment, while nine per cent expressed a preference not to accompany their child. The sex of the child (p = 0.33) or the fact that the parents were holders of a medical card (surrogate for disadvantage) (p = 0.08) did not affect parents' preference. However, parents of a single child had a higher preference (78 per cent) for accompanying their child than did parents with greater than one child in the family unit (66 per cent) (p = 0.0009). CONCLUSION: If given the choice, the majority of parents would prefer to accompany their child when receiving dental treatment. PMID- 15789986 TI - Necrotizing sialometaplasia: an important diagnosis--review of the literature and spectrum of clinical presentation. PMID- 15789987 TI - Orthodontic bonded retainers. AB - Retention is usually necessary following orthodontic treatment to overcome the elastic recoil of the periodontal supporting fibres and to allow remodelling of the alveolar bone. The degree of change is variable and largely unpredictable. Bonded lingual retainers have been shown to be an effective means of retaining aligned anterior teeth in the post-treatment position in the long term. Two basic designs of lingual bonded retainers are currently in use. Rigid mandibular canine to-canine retainers are attached to the canines only. They are effective in maintaining intercanine width but less so in preventing individual tooth rotations. Flexible spiral wire retainers are bonded to each tooth in the segment, their flexibility allowing for physiological movement of the teeth. This design is more effective at preventing rotation of the bonded teeth. Failure of bonded retainers may occur at the wire-composite interface, at the adhesive enamel interface or as a stress fracture of the wire. Failure of a retainer may lead to unwanted tooth movement. In many cases it will be possible to repair the appliance in the mouth. However, in some instances it will be necessary to replace the retainer. A disadvantage of fixed retainers is that they complicate oral hygiene procedures, and favour the accumulation of plaque and calculus. Despite this, the presence of a bonded retainer appears to cause no increase in incidence of caries or periodontal disease. Use of interdental cleaning aids is required to ensure adequate oral hygiene. PMID- 15789988 TI - Dental perspectives on neuropathic origin. AB - The most common cause of intraoral pain is odontogenic and rarely presents a diagnostic challenge. Pain in a tooth site area that is not dental or periodontal in origin may be difficult to diagnose and treat. Successful management of non odontogenic pain complaints is inherently dependent on taking a detailed 'pain history'. Treatments that are irreversible and potentially harmful to the underlying dentoalveolar structures must be avoided when the diagnosis is uncertain. Information gleaned from the pain history usually provides a provisional or working diagnosis, and this diagnosis must later be confirmed by specific testing (based on response to medication trials, imaging, laboratory tests, etc.). Patients who suffer from chronic pain conditions are likely to show comorbidity with other pain problems (e.g., neuropathic pain with background muscle discomfort or temporomandibular disorders and headache problems). Thus, in more complex patients, the various elements that comprise the chronic pain condition must be identified and managed, for adequate resolution of the symptoms. PMID- 15789989 TI - Record keeping. PMID- 15789991 TI - Over the threshold. PMID- 15789990 TI - Pawn in no-win game. PMID- 15789992 TI - New directions. PMID- 15789993 TI - Shower scene. PMID- 15789994 TI - Reflective practice: a meaningful task for students. AB - AIMS: To ascertain whether students found reflection to be a meaningful activity, whether there are perceived benefits associated with reflective practice and whether it is a valid process on which to assess the outcomes of a course relating to the competencies of specialist practice. METHOD: The attitudes of students attending a one-year degree course were examined using a focus group interview and a web discussion board. The evaluation of the focus group interviews and web-based discussion postings are discussed. FINDINGS: Reflective writing is considered a key component of portfolio assessment because it provides evidence of skills development and increasing clinical competence. There was no consensus on whether or not reflection is a meaningful activity. The students' learning style is pertinent to their perception of the usefulness of reflection. CONCLUSION: The findings indicated the scepticism with which some students approached the task, but they also identified that reflection had a positive impact on the practice of students more able to embrace the process in a meaningful way. There is scope to extend this work to examine the use of reflection in developing the growing body of practice knowledge that underpins nursing. PMID- 15789995 TI - Improving blood transfusion: a patient-centred approach. AB - More than three million blood components are transfused every year in the UK, providing life-saving and life-enhancing treatment to thousands of patients (McClelland 2001). However, transfusion error, resulting in the patient receiving the incorrect blood component, remains the largest risk related to transfusion in the UK. By developing accessible blood transfusion policies, auditable performance standards and training, and educational initiatives, nurses can increase compliance in high-risk areas of the transfusion process and reduce the potential for errors. PMID- 15789997 TI - Fertile new ground. PMID- 15789996 TI - Vascular access: indications and implications for patient care. AB - This overview of the vascular access devices available for adult patients explains how careful selection and management of the device can minimise the complications associated with infusion therapy. PMID- 15789998 TI - [The house in Solenn, welcoming suffering adolescents]. PMID- 15789999 TI - [Professor Marcel Rufo: The house at Solemm, "a unique place"]. PMID- 15790000 TI - [Knowing how to care for migrants and strangers in dangerous situations]. PMID- 15790001 TI - [Dialysis]. PMID- 15790002 TI - [Smoking and pregnancy]. PMID- 15790003 TI - [The masks unmasked]. PMID- 15790004 TI - [7 questions-answers on the diminution of care in nurses at risk for infections]. PMID- 15790005 TI - [Emla, the cream]. PMID- 15790006 TI - [Commission of relations with the customs and quality of care]. PMID- 15790007 TI - [See and do]. PMID- 15790008 TI - [Lack of communication and feelings of defeat]. PMID- 15790009 TI - The 'inside look' on correctional nursing: a unique nursing speciality. PMID- 15790010 TI - Emerging issue: Keeping patients and nurses safe. Nursing and musculoskeletal disorders. PMID- 15790012 TI - Evidence-based practice: A theory-based tobacco dependence treatment at an adolescent health clinic. AB - Tobacco is the leading cause of preventable death in the US. Most smokers report initiating smoking in adolescence when smoking is not perceived as a health risk. Assessment and education about the harm of tobacco use is critical to stemming the destructive behavior of smoking and avoiding lifelong addiction. This article describes a theory-based, nurse-run, adolescent-focused smoking assessment, education and intervention program. Results of the first 6 months of the program are described. PMID- 15790011 TI - Treatment and recovery. AB - Addiction is a chronic, progressive, fatal disease. It is a primary disease, not caused by any other disease process. It has recognizable symptoms that if left untreated, can lead to premature death, deterioration of physical conditions, social withdrawal and disintegration of all support systems. The family and co workers are affected as well. There is no cure, but we know that treatment works! Don't stand by while someone you care about needs help. Nurses helping nurses, let's take care of our own. PMID- 15790014 TI - [From obesity to type 2 diabetes in children and adolescents]. AB - Obesity has a prevalence of 15-16% among subjects aged 6-17 yr. in United States as well as in Europe. Another 10 to 15% of children and adolescents appear to be at risk of obesity. The presence of type 2 diabetes among adolescents in our country represents a challenge from both a screening and a therapeutic point of view. In addition to obesity, a positive familial history, puberty, ethnic susceptibility as well as conditions known to exhibit insulin resistance (acanthosis nigricans, dislipidemia, polycystic ovary syndrome) represent majors risk factors. Detecting subjects at risk among a large number of obese children appear to be a critical step. Therapy of type 2 diabetes requires as important means as those set for type I diabetes, taking into account the fact that both types of diabetes share the same vascular complications. PMID- 15790013 TI - [Enuresis and voiding disorders in childhood]. AB - Nocturnal enuresis is a common problem seen by the primary care physician. It is mandatory to distinguish between children having monosymptomatic nocturnal enuresis with normal daytime voiding habits and patients having polysymptomatic bed wetting (associated with urgency, frequency, or other signs of unstable bladder). Investigations and treatment of polysymptomatic enuresis are different than treatment of monosymptomatic nocturnal enuresis. A thorough and thoughtful history of voiding pattern is important to separate urge syndrome from organic causes of enuresis. Management of patients who have urge syndrome include general advices like regular voiding routine, physiotherapy, anticholinergic medication and prevention or treatment of urinary tract infections. If the nocturnal enuresis persists after the control of the voiding dysfunctions, treatment of nocturnal enuresis must be undertaken. PMID- 15790015 TI - [Hematuria and proteinuria in childhood]. AB - Hematuria and proteinuria are often the first signs of potentially severe kidney diseases. Investigations of a child with proteinuria +/- hematuria should start at the primary care physician office, and will permit to rapidly identify the most serious kidney diseases, such as the glomerulonephritis, but also to avoid excessive and costly investigations in patients with a benign condition such as orthostatic proteinuria. Isolated microscopic hematuria is also relatively frequently found during routine pediatric office visit. Secondary to a glomerulonephritis, it is often associated with proteinuria. Urologic causes should be excluded in case of isolated microscopic or macroscopic hematuria. PMID- 15790016 TI - [Acute dyspnea in children]. AB - Respiratory disease and acute respiratory difficulties are life threatening problems frequently met in paediatric medicine. Since parents often call their doctor first, telephone triage is important in the correct management of children with respiratory distress. On arrival in the office or the emergency department, a child with dyspnea should rapidly be assessed for signs of severity and respiratory compromise. Oxygenation and early initiation of specific treatment are priorities in the management. Only a simple and rigorous clinical process, based on the essential, will reach all these objectives without delay. As soon as the child's situation is stabilised, the doctor decides what are the appropriate modalities for transfer. This article aims at reviewing these different issues. PMID- 15790017 TI - [Recurrent lower respiratory tract infection in children: when and how should they be investigated?]. AB - Children with recurrent lower respiratory tract infections are often a challenge for their physicians. This article reviews the differential diagnosis of recurrent cough, bronchopneumonia and/or pneumonia in children and emphasizes on the necessity of preventing long-term complications of these infections. It also suggests a step-wise immunological work-up, which includes investigating a possible immune maturation delay or deficiency which could explain these symptoms. This work-up focuses on measuring antibody responses to pneumococci, followed by immunization when necessary, to rule out B cell dysfunction. Finally, this article also describes an upcoming study in Switzerland which will evaluate in 2005 the clinical and immunological outcome of these young patients. PMID- 15790018 TI - [Treatment of atopic dermatitis: practical approach]. AB - Atopic dermatitis (AD) is a chronic disease frequent in childhood. The treatment is based on regular moisturizing of the skin, information to the parents on the chronic course with recurrent flares, topical anti-infectious therapy for superinfections and colonization of the skin by staphylococcus aureus, and topical steroids. The immuno-modulatory macrolides (tacrolimus and pimecrolimus) represent a new alternative to topical steroids. These molecules are well tolerated, but theirs effects on the long-term are unknown. A food allergy may be responsible for a AD flare in up to a third of the cases, but the presence of an allergy should be demonstrated before the prescription of an elimination diet. AD is often the first manifestation of atopy: the physician should be aware of the future occurence of respiratory symptoms. PMID- 15790019 TI - [Antenatally detected hydronephrosis: practical approach for the pediatrician]. AB - Approximately 1% of the fetuses present some dilatation of their urinary tract in utero. More than 50% of these antenatally detected hydronephrosis will disappear spontaneously after birth. The other 50% comprises ureteropelvic junction obstruction, vesico-ureteral reflux and primary megaureters. Postnatal radiological evaluation (renal ultrasonography and VCUG) is performed in every infant with a significantly dilated renal pelvis (> 8 mm between 20 and 30 weeks or > 10 mm after 30 weeks in utero). Renal nuclear scan should be done in every child with significant/worsening post-natal hydronephrosis. Antibioprophylaxis will be started from birth to prevent urinary tract infection. Medical or surgical approach will be chosen in the light of the uroradiological exam results and the clinical progress. PMID- 15790020 TI - [Coercion in psychiatry: current knowledge and perspectives]. AB - Coercion is still frequently used in psychiatry and tends to be considered a necessity, when a patient at risk has lost his/her ability to judge. The aim of this article is to report and discuss current scientific knowledge on this topic. The different steps associated with coercion contribute, along with the illness, to the degree of stress experienced by the patient and might compromise his/her level of adherence to the treatment offered. Clinicians need to consider three dimensions: the coercion applied, the legal status and the subjective feeling of coercion. Pressure that is symbolically negative, positive or legal is exerted on the patients, thereby raising questions about the development of alternatives to the use of coerced treatment. The introduction of the advance directives to Swiss cantonal laws could give rise to different forms of collaboration that are more acceptable to patients. PMID- 15790021 TI - [Confidence in the drug industry]. PMID- 15790022 TI - [Alcohol and tobacco regulation in Great Britain and Cuba]. PMID- 15790023 TI - [The health status of the Pope]. PMID- 15790024 TI - [A new screening test for colorectal cancer?]. PMID- 15790025 TI - [The launch of a community-based facility for the renewed commitment of HIV prevention ]. PMID- 15790026 TI - [Molecular-genetic method of identification of drug-resistant Mycobacterium tuberculosis]. AB - Above two million people annually die worldwide of tuberculosis. There has been a serious uptrend in the tuberculosis morbidity observed during the recent decade in Russia. The growing prevalence of drug-resistant strains of pathogens is highly dangerous. A timely diagnosis reduces significantly the risk of epidemics. The use of molecular methods cuts the time of tuberculosis diagnosis, including its resistant forms, from several weeks to several days. The presented survey contains a discussion of modem methods used for identification of Mycobacterium tuberculosis resistance to the main anti-TB drugs. PMID- 15790027 TI - [Toxins of Vibrio cholerae]. AB - Surveyed in the paper are published data on properties, biological activity, genetic determinants and action mechanisms of recently known toxins produced by different strains of Vibrio cholerae irrespectively of their capacity for the synthesis of choleric toxin--the main virulence factor. Their possible importance both for the general clinical pattern of cholera provoked by cholerogenic agents and as independent virulence factors causing diarrhea without cholera is elucidated. The sets and levels of expression of additional toxins can differ for different pathogenic clones and they can correspondingly condition degrees of their epidemic and etiological safety. PMID- 15790028 TI - [Assembly and maturation of HIV-1--targets for new anti-HIV compounds]. AB - Much progress has been recently made in research of the final stages of the HIV-1 life cycle, i.e. of its assembly, gemmation and maturation. The virus was shown, in particular, to use widely cell mechanisms in its replication and assembly. The TSG 101 cellular endosomal sorting protein interacting with the p6 viral protein is necessary for gemmation. Cyclophilin and HP68 (cell proteins) needed for marphogenesis of the virus were identified. The recently obtained data on the interaction of Vif (a viral protein) and Apobec (a cell protein) showed that HIV 1 has an action mechanism overcoming the cell barriers. The "early" virus phenomenon, which is deprived of any mature structure or the ability to infect and does not contain mature Gag and Env proteins, illustrates that the proteolytic pressing of the Gag p55 precursor is not enough for the maturation of the virus and additional viral or cellular factors are needed for the virus to become infectious. Although the current antiviral therapy has been successful enough, it is far from being effective in all cases; one of the reasons is resistance to chemodrugs developing rapidly in patients. Fast mutations and exceptional plasticity of the viral genome (which helps the virus to develop rapidly resistance to drugs) belong to the major problems. The circulation of persistent virus variants has been quickly increasing. There is an urgent need in developing new antiviral drugs acting on new viral targets; progress in experimental virology would speed it up. Thus, new drugs can be created, which block the activity of Vif, that would make Apobec block the virus replication. Compounds can be developed, which block the interaction of cyclophilin and TSG101 with viral proteins. The recently described importance of cholesterol in the sexual transfer of viruses is expected to bring simple and inexpensive compounds destroying cholesterol in the mucous tunic of genitals into clinical use. The identification of additional factors needed for the maturation of the virus and for its becoming infectious can be a basis for the development of drugs blocking their packaging into virions. Future research is expected to define new targets for the chemotherapy of AIDS and to promote the designing of new chemodrugs. PMID- 15790029 TI - [Resistance of Russian isolates of Neisseria gonorrhoeae to fluoroquinolones]. AB - Fluoroquinolones still belong to the drugs of choice in the treatment of uncomplicated gonorrhea. At the same time, there have been more data on the spreading N. gonorrhoeae strains resistant to fluoroquinolones. A variety of mechanisms, like modification of the target of antibiotic's action (point mutations in genes gyrA and parC), a decreasing permeability of the bacterial cell membrane (amino-acid changes Por protein) and a growing efflux of antibiotic (mutations in the promoter or in the coding region of mtrR) mediate in the shaping resistance of the drugs. The MIC values for four fluoroquinolone-series antibiotics were determined and the gyrA, parC, por and mtrR genes were examined for resistance-responsible mutations in 32 studied clinical strains of N. gonorrhoeae. Strains with high resistance to fluoroquinolones were detected; 3 of them had no common changes in GyrA or ParC, however, amino acid changes and mutations were detected in Por protein and promoter or gene mtrR encoding region, respectively. The paper contains priority data on the detection (in Russia) of N. gonorrhoeae strains with high resistance to fluoroquinolones. Involvement of different mechanisms in the process of resistance shaping is discussed. The results are of practical importance for planning the antibacterial therapy of gonorrhoeae; they point out the need in regional testing of resistance in the N. gonorrhoeae population encountered in Russia. PMID- 15790030 TI - [Cloning and expression of Vibrio cholerae zonula occludens toxin (zot) gene in Escherichia coli]. AB - Two recombinant plasmids containing the cloned PCR-amplifled Vibrio cholerae zonula occludens toxin (zot) gene was constructed in orientation providing its transcription from lac-promoter. One of them contained also its own zot promoter. The third plasmid was obtained by subcloning a Vibrio cholerae DNA fragment including intact zot and ace (accessory cholera enterotoxin) genes. The expression levels of the cloned genes in Escherichia coli varied depending on a promoter type, host strain and culture conditions. The human intestinal cell line CaCo2 appeared to be a suitable model for assessing the biological activity of toxin preparations. The product of zot gene possessed a marked activity in respect to CaCo2 in spite of the lack of the molecule cleavage and transport of its toxic C-terminal part from alien host cells into the culture media. The constructed recombinant plasmids can be used as a source of molecular hybridization probes; and E.col transformants carrying those plasmids can serve in zot purification both for the scientific and practical purposes. PMID- 15790031 TI - [Production of recombinant fragment of VacA protein and development of non invasive method for the diagnosis of Helicobacter pylori infection]. AB - Vacuolizing toxin (VacA) Helicobacter pylori is an important factor of pathogenicity of Helicobacter pylori and a basic marker in the diagnosis of helicobacteriosis and related diseases. A coagulation-based diagnostic test system was elaborated for the detection of VacA in clinical samples. A fragment of vacA was cloned, for the purpose, in Escherichia coli and expressed in preparative quantities; the coded protein was purified and used in raising the diagnostic serum. The thus designed coagulating test-system was successfully tested under the modeling conditions with clinical samples. Therefore, the designed express method can be used for the invasion-free determination of VacA in patients with gastric and duodenal pathologies. PMID- 15790032 TI - [Regulation of the activity and synthesis of 3-desoxy-D-arabinoheptulose-7 phosphate synthase of Agrobacterium tumefaciens 2592]. PMID- 15790033 TI - [Molecular polymorphism of viral dsRNA of yeast Saccharomyces paradoxus]. AB - An analysis of 53 strains of yeast Saccharomyces paradoxus (YSP) of different geographic origins enabled us, for the first time, to find viral double-stranded RNA (L and M fractions) in YSP and to study natural polymorphism. As in the cultured Scerevisiae, the size of L dsRNA was constant (4.5 kb). The size of minor M dsRNA varied from 1.5 to 2.4 k.b. In YSP, we determined 7 types of M dsRNA (M1-M7), which were not connected with the source of isolation or geographic origin of the host strains. PMID- 15790034 TI - Physician workforce: colleagues of the future. PMID- 15790035 TI - American health care in decline. PMID- 15790036 TI - A brief history. PMID- 15790037 TI - Mercer University School of Medicine: a story about promises made, promises kept. PMID- 15790038 TI - The Medical College of Georgia: the state's oldest medical school has a new lease on life. PMID- 15790039 TI - From whence they came: medical education. PMID- 15790040 TI - The government wants me to buy what? Health information technology in the physician's office. PMID- 15790041 TI - Choosing a field of medicine: a medical student's perspective. PMID- 15790042 TI - Morehouse School of Medicine. PMID- 15790043 TI - "It was the best of times; it was the worst of times". PMID- 15790044 TI - Liver hydatid disease: morphological changes of protoscoleces after albendazole therapy. AB - BACKGROUND: Postoperative recurrence of the liver hydatid disease befalls approximately 10-30% of patients. Preoperative or postoperative therapy with albendazole in single therapeutic protocol (800 mg/d, within 28 days) indicated the need to evaluate the hydatid cyst liquid protoscoleces viability. Morphological changes of protoscoleces following the treatment with drugs are not well known. AIM: To estimate the viability of protoscoleces after preoperative or postoperative albendazole therapy, and their ability for cystic metamorphosis. METHODS: A prospective, randomized clinical trial included 30 patients with liver hydatid disease, treated with albendazole and surgically (I group), and 30 patients in the control group treated only surgically (II group). The concentration of albendazole and its active metabolite albendazole sulphoxide in the cysts contents were determined using HPLC. Estimation of protoscoleces viability was based on the established micromorphologic criteria, and compared between the patients treated with albendazole, and the patients treated only surgically. Biological assessment of the viability was performed on protoscoleces with uncertain signs of the disturbed viability (unchanged structure, evaginated, without movements) using intraperitoneal injection of 1 ml of protoscoleces prepared suspension to AO type of rats. RESULTS: The concentration of albendazole in cysts' contents ranged from 0 to 64.9 microg/ml, and of its active metabolite from 0.5 to 40.8 microg/ml. The presence of fully viabile protoscoleces in the albendazol treated patients was significantly lower than in the control group. A significant difference was noticed in the presence of disintegrated protoscoleces without movements in the albendazol treated group, compared to the control group. Biological assessment of the viability showed incapability of these protoscoleces for cystic metamorphoses. CONCLUSION: Low viability of parasites due to medicamentous therapy is very useful and important to surgeons, because the fertility of cysts is lower, and the risk of the disease recurrence is reduced. PMID- 15790045 TI - [Resistance of catheter-associated urinary tract infections to antibacterials]. AB - INTRODUCTION: Catheter-associated urinary tract infections (CAUTI) are the most common nosocomial infections. The worldwide data show the increasing resistance to conventional antibiotics among urinary tract pathogens. AIM: To evaluate the adequacy of initial antimicrobial therapy in relation to the antimicrobial resistance of pathogens responsible for CAUTI in Clinical Center of Banja Luka. METHODS: A retrospective study on major causes of CAUTI, antibiotic resistance and treatment principles was conducted at four departments of the Clinical Center of Banja Luka from January 1st, 2000 to April 1st, 2003. RESULTS: The results showed that 265 patients had developed CAUTI. The seven most commonly isolated microorganisms were, in descending order: E. coil (31.0%), Pseudomonas aeruginosa (13.8%), Proteus mirabilis (12.9%), Gr. Klebsiella-Enterobacter (12.3%), Enterococcus spp. (5.2%), Pseudomonas spp. (4.3%), Serratia spp. (4.0%). The most common pathogens were highly resistant to ampicillin (64-100%), gentamycin (63 100%), and trimethoprim-sulfamethoxazole (68-100%), while some bacterias, like Pseudomonas aeruginosa and Serratia spp. showed rates of ciprofloxacin resistance as high as 42.8% and 72.7%, respectively. In 55.5% of the cases, the initial antibiotic therapy was inadequate, and was corrected latter on. There were no standard therapeutic protocols for this type of nosocomial infections. CONCLUSION: The results of this study emphasized an urgency of the prevention and introduction of clinical protocols for better management of CAUTI. Treatment principles should better correspond to the antibiotic sensitivity of uropathogens. PMID- 15790046 TI - [Variability in interpretation of static renal scintigraphy findings]. AB - BACKGROUND: Static renal scintigraphy with 99mTc-DMSA is a sensitive method for evaluation of any reduction of cortical mass function. There is not enough reliable references in the literature on scintigraphic reports in terms of objectivity and standardization. The aim of this paper was to assess the differences in interpretation of routine renal scintigraphic findings by nuclear medicine specialists from various institutions (interobserver variability). METHODS: Both interobserver variability and agreement in the interpretation were evaluated in two groups. Six observers from four different institutions (group A), and three observers from the same institution (group B) interpreted independently 60 static renal scintigraphy findings obtained by analogous imaging using x-ray films in four standard projections. Using at least two options, seven parameters of renal scintigraphy protocols were analysed: size (1), uptake (2), outlines (3), scars (4), focal changes (5), relative function (6), and conclusions (7). RESULTS: The interpretations of scintigraphic findings were in accordance in items 1 to 7 within the group A: 62, 42.5, 45, 9, 47, 52, and 34 percent respectively, and within the group B: 72, 55, 59, 22, 62, 60, and 41 percent, respectively. The conformity was more significant in the findings observed in group B as opposed to group A, which was statistically highly significant, especially for the category of scars (p < 0.01). Significant variability was also observed by six observers in all the parameters of scintigraphic protocol, especially, again, in the scars analysis (p < 0.01). CONCLUSION: Insignificant degree of accordance, especially in the assessment of scars necessitates the standardization of criteria and terminology in order to interprete the renal scintigraphy with more accuracy and objectivity. PMID- 15790047 TI - [Fine-needle aspiration cytology in Hodgkin's lymphoma]. AB - BACKGROUND: Cell composition of tumor tissue in Hodgkin's lymphoma is considered to be specific. According to some authors, precise cytomorphological analysis of the specimen obtained by fine needle enables the diagnosis of the histologically well-defined form of lymphoma in a high percentage of cases. However, other authors consider the precise diagnosis of HL based on cytological analysis of a lymph node puncture specimen extremely difficult. The aim of this study was to a assess the diagnostic accuracy of cytomorphological analysis, as well as the possibility of application of this method in establishing the preliminary or even final diagnosis of this form of lymphoma. METHODS: Thirty four samples were analyzed, obtained by fine needle aspiration, in patients examined and treated at the Institute of Hematology, Clinical center of Serbia, Belgrade. Smears were stained by May-Grunvald-Giemsa and Papanicolau methods. RESULTS: The finding of typical multinucleate Reed-Sternberg cells in the smears of lymph node puncture specimen was diagnostically significant, as well as the finding of mononuclear variances of the neoplastic cells of anunusual morphology (Hodgkin's cells). The cell environment in the background of the above-mentioned tumor cells was heterogeneous. It was composed of small lymphocytes, plasma cells, eosinophils and reticular cells. These cell forms, found in cell preparations, resembled those seen in the tissue sections of the relevant extirpated lymph nodes. The comparison of the cytological findings in the fine needle specimen to the subsequently established histological diagnoses of HL revealed a high correlation of 85.2%. CONCLUSION: Cytological examination of fine needle lymph node specimens is still significant for the diagnosis of HL in spite of the development of more sophisticated methods used for verification of the diagnosis of this neoplasm. PMID- 15790048 TI - [Factor V Leiden, FII G20210A, MTHFR C677T mutations as risk factors for venous thrombosis during pregnancy and puerperium]. AB - BACKGROUND: Venous thrombosis is the most common cause of obstetric morbidity and mortality during pregnancy and puerperium. The incidence of pregnancy-associated venous thrombosis varies from 1 in 1000 to 1 in 2000 deliveries. Factor V G1691A (FV Leiden), FII G20210A and MTHFR C677T mutations are the most common genetic risk factors for thromboembolism. The aim of this study was to establish the presence of these risk factors in a group of women with an episode of deep venous thrombosis during pregnancy or puerperium. METHODS: The study was carried in a group of 45 women with the first episode of deep venous thrombosis during pregnancy or puerperium. The patients with antiphospholipid antibodies, antithrombin III, protein C or protein S deficiency, and autoimmune and malignant diseases were excluded from the study. FV Leiden, FII G20210A, and MTHFR C677T mutations were detected by polymerase chain reaction, followed by digestion with specific restriction enzymes. RESULTS: Twenty heterozygous carriers of the FV Leiden mutation and one homozygous carrier were detected, which represents the frequencies of 44.4% and 2.2%, respectively. For the FII G20210A mutation, six heterozygous carriers were identified, giving the frequency of 13.3%. The MTHFR C677T mutation was observed in 31 patients (22 heterozygous and 9 homozygous carriers) which represents the frequencies of 48.9% and 20%, respectively. CONCLUSION: Our study suggested that the obligatory testing for FV Leiden and FII G20210A mutations was strongly recommended in women with history of venous thrombosis during pregnancy and puerperium. We found a slight effect of MTHFR 677T allele, but it should be considered in association with other risk factors. PMID- 15790049 TI - [Histopathologic investigations of nifedipine-induced gingival hyperplasia in Wistar rats]. AB - BACKGROUND: Nifedipine, a calcium channel blocker, used in the therapy of cardiovascular diseases, can induce gingival overgrowth. The aim of the study was to assess the influence of nifedipine on the deeper periodontal tissues of the experimental animals concerning connective tissue and blood vessels proliferation. METHODS: The research was performed on Wistar rats, and they received the precise doses of nifedipine water suspension while the changes were followed up in pre-determined time intervals. The analyses of samples of tissues were done on the cross-sections of papilla from the top to the bottom on five levels. RESULTS: Considerable level of changes was observed in the area of dental alveolar edge and in the gingivo-dental interspace. Epithelial hyperplasia and inflammatory subepithelial infiltration of lymphoplasmocyte type were also present. Alveolar bone destruction, the increased number of osteoclasts and periodontal destruction were found, as well. CONCLUSION: The results of this study showed a significant gingival hyperplasia in the experimental animals which received high doses of drug during extended period of time. PMID- 15790050 TI - [Use of high-dosage chemotherapy with autologous hematopoietic stem cell transplantation as a first-line therapy for the patients with poor-prognosis testicular tumors]. AB - BACKGROUND: High-dose chemotherapy followed by hematopoietic stem cell support can be used as a first-line treatment in patients with germ-cell tumor (GCT) with poor prognosis. Long-term survival rate is attained in 50% of these patients. The aim of this paper was to present the experience at the Department of Hematology, Military Medical Academy, with high-dose cytostatic therapy as first-line chemotherapy in GCT patients with poor prognosis. METHODS: Between 1997 and 2003, five patients with high-risk germ-cell tumors were treated with high-dosage chemotherapy followed by an autologous stem cell transplantation. All the patients were with non-seminomatous germ-cell tumors with mixed histology, and one was with extragonadal retroperitoneal germ-cell tumor. RESULTS: The follow-up period ranged from 8 to 33 months. Three patients achieved complete remision, two patients only partial remision, and one was not followed-up. One patient was with residual tumor resection, using retroperitoneal lymphadenectomy, after autologous stem cell transplantation. All the patients were treated according to standard protocols. CONCLUSION: Early high-dose chemotherapy associated with hematopoietic stem cell support as a first-line treatment in the patients with germ-cell tumor with a poor prognosis, represented an efficient treatment modality. PMID- 15790051 TI - [Immunopathogenetic mechanisms of chronic dental periapical lesions]. PMID- 15790052 TI - [Effect of selenium on health protection and development of disease]. PMID- 15790053 TI - [Papillary muscle rupture due to acute myocardial infarction followed by cardiogenic shock, pulmonary edema, and acute renal failure]. AB - AIM: The case of successfull surgical treatment of anterolateral papillary muscle rupture due to acute myocardial infarction with cardiogenic shock, pulmonary edema and acute renal failure. CASE REPORT: A 62-year old male from Belgrade with chest pain, hypotension and a new heart murmur refused hospitalization at the Military Medical Academy. On the third day of his illness he was readmitted to MMA as an emergency due to hemoptysis. Examination revealed mitral valve anterolateral papillary muscle rupture. The patient, with signs of cardiogenic shock and acute renal failure, was immediately operated on. The surgery was performed using extracorporeal circulation. An artificial mitral valve was implanted, and myocardial revascularization accomplished with one venous graft of the left anterior descending artery. On the second postoperative day, hemodialysis was carried out due to acute renal failure. On the 28th postoperative day, the patient was discharged from the hospital being hemodynamically stable with normal renal function and balanced anticoagulation. The case is interesting in terms of unrecognized papillary muscle rupture that led to the development of cardiogenic shock, hemoptysis and acute renal failure. CONCLUSION: Papillary muscle rupture is a fatal complication of acute myocardial infarction. Early recognition and urgent surgical intervention were life-saving in the case of complete papillary muscle rupture. Surgical treatment, regardless of high risk, is the procedure of choice. PMID- 15790054 TI - [Acute leg compartment syndrome after exertion]. AB - A case of a 22-year old soldier, with a history of pain in the leg during heavy exercise, which desisted at rest, was presented. One day before admission, the patient had felt an extreme exertion-induced pain in his right leg which had not lessenned at rest. At the same time, the patient noticed persistent severe leg edema. On physical examination, the intracompartmental pressure was 62 mmHg (> 30 mmHg). The patient was urgently operated on, and fasciotomy according to Mubarak was used. At second surgery, the debridement of the muscles of the posterior group of the leg, and the evacuation of hemathoma from the anterior and lateral group of the right leg muscles were perfomed. Postoperative recovery was uneventful. Fasciotomy wounds were closed within 14 days of the surgery. The complete physical treatment was done. Follow-up examinations 1, 3, and 6 months afterwards were satisfactory. The soldier completed his compulsory military service without any sequelae. Laboratory results were normal. Overlooked, unrecognized or surgically untreated compartment syndrome can cause severe damage, including even the loss of the extremity. PMID- 15790055 TI - [Therapy of acute hepatitis C infection]. AB - BACKGROUND: Hepatitis C viral infection is a major cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. The progression of acute to chronic infection occurs in 50-90% of cases. There is no standard therapy for acute HCV infection. Comparative studies are required to verify the optimal doses, dosage schedules and the treatment duration, and to establish the optimal treatment for acute hepatitis C. Recent reports have demonstrated that early application of interferon alpha was a treatment of choice for acute HCV infection. The addition of ribavirinin in the treatment of acute HCV infection, and HCV genotype, did not improve the end-of-treatment responses. It is important to consider the treatment of acute HCV infection before it progresses to chronic state. CASE REPORT: Beneficial effect of interferon therapy in a patient with acute hepatitis C is presented. Early treatment with 3 MIU interferon alpha, three times a week, within six-months, resulted in the normal serum aminotransferases, and good virological response in our patient. CONCLUSION: Interferon therapy significantly increased the probability of obtaining normal serum aminotransferases and undetectable HCV RNA, following acute HCV infection. PMID- 15790056 TI - President's page: Continuing the struggle. PMID- 15790057 TI - History of a public health emergency. PMID- 15790058 TI - Clinical-trials registry. PMID- 15790059 TI - New Jersey Medicine interviews Eric Munoz, MD, MBA, assemblyman and surgeon: Expert in health policy provides blunt answers. PMID- 15790060 TI - Dr. Coates and the Know Nothings. PMID- 15790061 TI - Setting goals that work. PMID- 15790063 TI - A conservative case for universal health insurance. PMID- 15790062 TI - A twenty-six-year-old Trinidadian woman with dizziness, nausea, and vomiting. PMID- 15790064 TI - Improving medical practice and the economy through universal health insurance. PMID- 15790065 TI - Controversies in mammography. AB - Concerns about screening mammography include questions of efficacy, high recall rates, false positives, and age at which to institute annual screening. Annual screening mammography can decrease breast cancer mortality by 45% in women over fifty and 23% in women between forty and fifty years of age. Patient recall rates and accuracy of interpretation vary among radiologists. Suggestions for improving accuracy of mammographic interpretation include continuing education with emphasis on quality of interpretation, computer-assisted detection, double reading, increased volume per reader, and performance-related skills testing. Having fewer radiologists reading more mammograms may result in decreased patient access to mammography services. Poor reimbursement for mammography and high prevalence of breast cancer-related litigation are disincentives for radiologists to provide mammography services; these issues must be addressed to ensure patient access to mammography. The public must be educated so that reasonable expectations on the benefits and limitations of mammography will develop. PMID- 15790066 TI - Heading-off anthrax at Hamilton, New Jersey: events of October 2001. PMID- 15790067 TI - The USS New Jersey. PMID- 15790069 TI - Mechanism of vancomycin resistance in methicillin resistant Staphylococcus aureus. AB - A collection of laboratory mutants and clinical MRSA strains, additionally exhibiting resistance to glycopeptide antibiotics, was studied in detail. The nature of resistance to glycopeptides was found to be different from that existing in vancomycin resistant (VR) enterococci. The mutants produced abnormal murein in which the level of highly oligomeric muropeptides was drastically reduced. Biochemical and genetic analyses of Penicillin Binding Proteins (PBPs) showed inactivation of PBP4. Changes in other PBPs were not apparent, except for PBP2a that was inactivated in the highly VR mutant VM. Transposon inactivation of the pbpB gene and several other genes involved in synthesis of staphylococcal peptidoglycan all caused dramatic reduction of glycopeptide resistance in the staphylococcal mutants. While inactivation of PBP2a slightly increased the levels of glycopeptide resistance, a combination of vancomycin or teicoplanin with beta lactam inhibitors, chosen on the basis of their relatively selective affinities for individual staphylococcal PBPs completely inhibited the expression of glycopeptide resistance in MRSA. Glycopeptide antibiotics caused a virtually complete inhibition of cell wall turnover and autolysis and massive overgrowth of cell wall material in the glycopeptide resistant mutants. Bacteria were able to remove quantitatively glycopeptide molecules from the growth medium, and sequestered antibiotic could be recovered in biologically active form from the purified cell walls. These observations and the results of the vancomycin binding studies suggest alterations in the structural organization of the mutants' cell wall such that access of glycopeptide molecules to the sites of wall biosynthesis is blocked by steric hindrance. PMID- 15790068 TI - The CPC in the December issue. PMID- 15790071 TI - Identification, cloning and expression of Pseudomonas aeruginosa Ps-x putative urate oxidase gene in Escherichia coli. AB - In a previous study we reported for the first time the isolation and characterization ofurate oxidase enzyme from Pseudomonas aeruginosa. In this work we isolated and cloned a 1.350 kilobase DNA fragment that encode a putative urate oxidase gene from the genomic library of P. aeruginosa Ps-x. The nucleotide sequence of the cloned DNA insert revealed an open reading frame that encodes a protein of a molecular weight of 54.0 kDa. The cloned DNA fragment showed an uricolytic activity when expressed in E. coli DH5alpha. Surprisingly, the nucleotide sequence of the cloned gene showed more than 99% identity to the gene encoding hypothetical protein of P. aeruginosa PAO1. Moreover, the sequence of the cloned gene was closely similar to the corresponding uricase gene of Cellulomonas flavigena (44% similarity), but showed lower similarity values to that of Bacillus sp. BT-90 (24% similarity), Candida utilis (24% similarity). Interestingly, the isolated uricase gene showed closer similarity to uricase from yeast-like symbiotic fungi Beauveria bassiana (35%), Tolypocladium inflatum (29%), Paecilomyces tenuipes (27%) and Cerataphis fransseni (24%). PMID- 15790070 TI - Genetic diversity among Bacillus anthracis, Bacillus cereus and Bacillus thuringiensis strains using repetitive element polymorphism-PCR. AB - Repetitive element polymorphism-PCR (REP-PCR) is one of the tools that has been used to elucidate genetic diversity of related microorganisms. Using the MB1 primer, REP-PCR fingerprints from 110 Bacillus strains within the "B. cereus group" have identified eighteen distinct categories, while other more distantly related bacterial species fell within six additional categories. All Bacillus anthracis strains tested were found to be monomorphic by fluorophore-enhanced REP PCR (FERP) fingerprinting using the MB1 primer. In contrast, other non- B. anthracis isolates displayed a high degree of polymorphism. Dendrogramic analysis revealed that the non- B. anthracis strains possessing the Ba813 chromosomal marker were divided into two clusters. One of the clusters shared identity with the B. cereus strains examined. PMID- 15790072 TI - Murein-hydrolyzing activity of flagellin FlaA of Listeria monocytogenes. AB - In this preliminary report we show that a 29 kDa surface protein of Listeria monocytogenes EGD removed from cells with 4 M LiCl has peptidoglycan (murein) hydrolyzing activity, as revealed by zymographic analysis using Bacillus subtilis murein and heat-killed Micrococcus luteus cells casted in the gel. Following two dimensional electrophoresis, the protein was electroblotted to PVDF membrane and its identity (FlaA) was revealed by sequencing. Peptidoglycan hydrolysing activity of FlaA purified by FPLC on Mono-S Sepharose against labelled Escherichia coli murein was demonstrated. PMID- 15790073 TI - Role of pencillin binding proteins in pencillin allergy. AB - All beta-lactam sensitive bacteria contain enzymatic penicillin binding proteins (PBPs), which are membrane-bound enzymes and targets of beta-lactam antibiotics. In this work evaluation of the significance of PBPs in immune response to benzylpenicillin was presented. 35 patients with allergic reactions to penicillin and 17 subjects without penicillin allergy, but exposed, were studied. Proliferative T-cell responses to benzylpenicillin, penicillin and PBPs conjugates (Pc-PBPs) from E. coli, K. oxytoca, S. aureus, S. epidermidis or serum protein (Pc-S) were measured. Although each allergic individual responds to Pc PBPs in several different ways, specific proliferation of T lymphocytes with Pc PBPs from bacterial membranes was significant higher than with Pc or Pc-S. This observation gives us a real insight into the causes of the drug allergy: individual allergic reaction and susceptibility to the drug is in strict correlation with bacterial infection. It seems likely that acylation of PBPs could be the trigger for primary immune response to the hapten benzylpenicillin. PMID- 15790074 TI - Bacteria forming a resident flora of the skin as a potential source of opportunistic infections. AB - Along with progress of medicine, contribution that opportunistic bacteria make in nosocomial infections increases. Coagulase-negative staphylococci are these multiresistant strains which often cause this kind of infections. But more and more frequently other genera of bacteria are isolated. The main source of them is first and foremost the hospitalized patient's endogenous flora e.g. from their skin, because transmission of bacteria from this source is very effective. Analysis was concerned with bacteria that were recovered repeatedly from the skin of young, healthy men during period of five months. Composition of resident bacteria, after removing transients was evaluated. The number of microorganisms per 1 cm2 patients' skin was a constant value but different for each patient. Newly composed media enabled exact isolation and qualitative analysis of all groups of expected bacteria. Isolated microorganisms represented three main groups: sensitive to novobiocin staphylococci, microaerophilic rods from Propionibacterium genus and corynefonn bacteria. Aside from quantitative differences in total bacteria number, significant differences in contribution of aerobic and anaerobic flora living on patient skin were observed. A persistent although not predominant population occurring on the skin of all patients in similar number (average 2%), were coryneform bacteria. They mainly belonged to the Corynebacteriumn genus, and 84.7% of them were the lipophilic species. These bacteria deserve special attention because among such species isolated from nosocomial infections, multiple antibiotic resistance of unknown origin was described. PMID- 15790076 TI - Effect of age on the fatty acid composition of the Bacillus subtilis PO270 isolated from wheat rhizosphere. AB - The changes of the composition of growing medium and the fatty acid composition of Bacillus subtilis PO270, a bacterium isolated from the wheat rhizosphere, was evaluated during different phases of growth. During growth alkalinity reaction of medium was observed and in late stationary phase of growth the release of proteins and phenolic acids from cells was observed. Twenty six fatty acids were detected. The most prominent fatty acids found in bacterial cells were 15:0 a, 15:0 i, 17:0 alpha and 17:0 i. Depending of a phase of bacterial growth, their contents varied from 86.5 to 88.9% of total fatty acids. The remaining fatty acids identified, including regular saturated and monounsaturated as well as iso- and anteiso-branched, 2- and 3-hydroxylated, cyclopropane and odd-numbered derivatives, were present in minor amounts. We have demonstrated that the fatty acid composition of this bacterium changes greatly in different growth phases. These structural changes represent re-arrangement of membranes, which keeps the bacterial cell fit during growth and counteracts the effects of the changing environment. PMID- 15790075 TI - A correlation matrix of alkaline phosphatase and sporulation in diazotrophic cyanobacteria and its thermo-tolerant mutant. AB - Alkaline phosphatase (phosphomonoesterase i.e. PMEase) activity in heterocystous cyanobacteria Anabaena flos-aquae, Nostoc calcicola, Calothrix brevissima, Scytonema javanicum and Hapalosiphon intricatus is known to be temperature and pH dependent. Maximum level of enzyme activity was recorded at either 35 degrees C or 37.5 degrees C. Also, the cell bound phosphomonoesterase enzyme was shown to exhibit pH optima of 10.0 or 10.2. A thermo-tolerant (tr) mutant isolated after MNNG (N-methyl-N'-nitro-N-nitrosoguanidine) mutagenesis in Calothrix brevissima exhibited 10 degrees C higher temperature optima and comparatively high pH optima (pH 10.4) for phosphomonoesterase enzyme. The mutant grew with a maximum growth rate (k) at 50 degrees C. Activation energy (Ea) for cyanobacterial strains was in a narrow range between 45 to 52 kJ mol(-1). A little variation in temperature and pH optima was also observed in phosphomonoesterase activity of Calothrix brevissima and its thermo-tolerant mutant while utilizing various organic phosphates as substrate what indicated the substrate dependence temperature and pH optima. Cyanobacterial strains grown at their respective temperature and pH optima differentiated spores less frequently though, coupled with early initiation of spore. PMID- 15790077 TI - The role of microorganisms in dispersion of thallium compounds in the environment. AB - Thallium is a highly toxic element and very rarely studied in the context of environmental hazards connected with zinc and non-ferrous metal industry. Microorganisms naturally existing in post-flotation and smelt wastes can participate in thallium release from waste deposits and can contribute to its dispersion in the environment. Twenty-one isolates were obtained from wastes of a non-ferrous smelter in Southern Poland characterised by high heavy metal contamination. Ten isolates showed high activity in thallium leaching from wastes (post-flotation and smelt wastes) as well as from pure thallous sulphide. Additionally, cadmium and lead were bioleached from wastes. The isolated bacteria indicated thallium resistance at a concentration up to 100 mg/l and some of them were able to survive in good condition at a concentration of up to 4 g/l. The same bacteria were isolated from rivers and wastewater in this region. A preliminary characterisation of isolates was performed. It was shown that some petroleum products i.e. asphalt-base crude occasionally used for waste immobilisation at the edge of pond or flotation surfactants partially stopped the activity of sulphide oxidising bacteria. PMID- 15790078 TI - Antimicrobial susceptibilities of Listeria monocytogenes strains isolated from 2000 to 2002 in Poland. AB - Minimal inhibitory concentration (MIC) of ampicillin, penicillin, gentamicin, erythromycin, clarithromycin, sulfisoxazole and trimethoprim for 73 Listeria monocytogenes isolates from clinical material samples, food and environment was determined using broth microdilution method. With exception of the sulfonamide all antibacterial agents were active against all tested strains. Resistance to sulfisoxazole (MIC > or = 512 microg/ml) was detected in case of 30.1% of isolates including 5 of 14 human strains. PMID- 15790079 TI - Characteristics of bacterial strains inhabiting the wood of coniferous trees. AB - The presented studies embraced samples of wood chips from coniferous trees which contained layers of duramen, alburnum and bark. Microbiological analysis involved qualitative and quantitative determination of bacterial flora inhabiting the studied wood material. The wood chips were found to contain primarily species belonging to the genera Bacillus and Pseudomonas. The presence of the potentially pathogenic species Bacillus cereus 1, Sphingomonas paucimobilis, Aeromonas salmonicida and Chryseomonas luteola was also demonstrated. PMID- 15790080 TI - Molecular epidemiology of Salmonella enterica serovar Enteritidis strains by pulsed-field gel electrophoresis isolated in the Slovak Republic. AB - A total 44 isolates of S. enterica serovar Enteritidis (S. Enteritidis) belonged to three different phage types (PTs) 9a, 13a, 25 were analyzed by the technique of pulsed-field gel electrophoresis (PFGE). Thirty and three strains were from two outbreaks (central and southern regions of the Slovak Republic PTs 13a, 25) and 11 isolates were sporadic isolates (PT9a). These isolates showed two different patterns in PFGE with XbaI digestion. Strains of PT13a generated PFP A and isolates of PT25 showed uniform PFP B. Nine sporadic isolates of PT9a belonged to PFP A and two isolates to PFP B. The PFPs A and B were differed by only two bands. The distribution of XbaI profiles did not corresponded with PTs. We conclude that the close genetic similarity observed between epidemiologically unrelated and outbreak-related isolates of S. Enteritidis suggest clonal relationship of these isolates. PMID- 15790081 TI - Radiologic case study. Spontaneous osteonecrosis of the knee reclassified as insufficiency fracture. AB - Three questions need to be answered. First, does spontaneous osteonecrosis of the knee in fact exist? The answer is "probably," but much more rarely than initially believed. In five of the authors' most recent MRIs in patients with clinically diagnosed or radiographically suggested spontaneous osteonecrosis of the knee, only one patient's images fulfilled the criteria for spontaneous osteonecrosis (Figure 3). Three of the five had subchondral changes of either insufficiency fractures or true fractures, while one patient had equivocal findings. Second, will the actual diagnosis influence the treatment? The answer appears to be "no" because early subchondral fractures and early spontaneous osteonecrosis of the knee are treated conservatively and only the later changes of subchondral surface collapse are treated more aggressively with joint replacement. Third, is there any method of halting the progression of early subchondral changes before they progress into collapse, flattening, and loose-body formation? The answer appears to be "maybe" as non-weight-bearing conservative therapy has not consistently prevented progression of the disease. Whether new arthroscopic techniques can provide the necessary intervention to halt osteoarthritic sequelae remains unanswered. Thus, it appears most patients with what has been called spontaneous osteonecrosis of the knee actually have subchondral insufficiency fractures. The histologic findings of avascular areas in biopsy specimens appear to be a secondary phenomenon and not the primary cause. If diagnosed early enough, conservative therapy may lead to healing without subsequent collapse and fragmentation of the subchondral bone. PMID- 15790082 TI - Vioxx and Merck: a preventable situation? PMID- 15790083 TI - Limited lateral acromioplasty for rotator cuff surgery. PMID- 15790084 TI - Precise positioning of unreamed solid nails in short distal libial fragments with percutaneous K-wires. PMID- 15790085 TI - Hip impingement: the role of arthroscopy. PMID- 15790086 TI - "Collateral damage": antibiotics and the risk of Clostridium difficile infection. AB - Patients undergoing orthopedic surgery are at increased risk for infection, and antimicrobial use continues to be required. Antimicrobial selection, however, is an important consideration given the increasing incidence and severity of C. difficile infection described in the literature. When choosing antimicrobials for prophylaxis and treatment, evaluate patients for risk factors that may predispose them to C. difficile infection. Patients receiving multiple antibiotics or broad spectrum antibiotics, women, patients with concurrent proton pump inhibitor use, and those with renal failure are at increased risk. Choice of antibiotics should be evaluated for their potential or likelihood to cause C. difficile infection. When a number of these risk factors are present, avoiding the use of high-risk antibiotics may be warranted. PMID- 15790087 TI - Meniscus repair update: current concepts and new techniques. PMID- 15790088 TI - Rapid prevention of vertebral fractures associated with osteoporosis. AB - Osteoporosis affects postmenopausal women and patients on glucocorticoid therapy. Fractures are the most devastating outcome. Patients who experience an osteoporotic vertebral fracture are at substantial risk of experiencing another within 1 year. Risk can be reduced rapidly with antiresorptives. Risedronate reduced the risk of vertebral fracture in patients with post-menopausal or glucocorticoid-induced osteoporosis after 1 year by up to 71% in prospective studies. In post hoc analyses, significant reductions in clinical vertebral fractures were demonstrated after 6 months with risedronate and 1 year with alendronate and raloxifene. Rapid reduction in fracture risk is achievable with the potent therapeutic agents available. PMID- 15790089 TI - The effect of nonsteroidal anti-inflammatory agents on spinal fusion. AB - A large body of information suggests NSAIDS have a negative impact on the healing of bone. Although each clinical healing scenario presents a slightly different level of challenge, the healing of a posterolateral spinal fusion is one of the most difficult challenges in bony healing. Clinically, this results in a relatively high rate of nonunions using traditional fusion techniques. Spinal fusion models have confirmed NSAIDS have a definite inhibitory effect on healing of the fusion. Although data are limited, it appears this effect is most severe when NSAIDS are administered in the early postoperative period. Moreover, the effect may be worse with certain types initial inflammatory, subsequent reparative, and final remodeling phases. Because of the anti-inflammatory activity of NSAIDS, one might assume their effects would be worse when administered in the inflammatory phase. Indeed, the study by Riew et al suggests the inhibitory effects are more significant when NSAIDS are administered earlier following fusion. Other studies conducted with non-spinal models also suggest early administration of NSAIDS results in greater inhibition of bone formation (Goodman et al). Unfortunately, the length of the inflammatory phase in humans is not well known. This leaves the clinician unsure about the safe time to allow resumption of NSAID usage clinically. It appears likely NSAID use following a spinal fusion procedure will increase the rate of pseudarthrosis. The literature suggests that avoidance of NSAIDS in the postoperative period may avoid nonunion. Additionally, we propose that chronic NSAID usage should be addressed in a similar manner to cigarette smoking. While neither are absolute contraindications to elective spinal fusion, patients should be counseled to discontinue the use of NSAIDS in the peri- and postoperative period to maximize their chance for a successful fusion. PMID- 15790090 TI - Sacral resection and reconstruction for tumors and tumor-like conditions. AB - Sacrectomy is intended to resect aggressive local and life-threatening disease. Reconstruction can be difficult. This article reports on the use of sacrectomy for tumor and tumor-like conditions. Procedures included 9 partial, 7 subtotal, 7 hemi-, and 4 total sacrectomies. Of 22 patients with primary malignant disease, 64% were alive without evidence of disease. The local recurrence rate was 27%. Thirty-two percent developed metastatic disease and 32% died from progressive disease. The complication rate was 33%. Tumor control was achieved in 73%. Postoperative complications occurred in 33%. Sacral resection remains a challenging operation. PMID- 15790091 TI - Clinical and cytogenetic findings in 14 patients with madelung anomaly. AB - Madelung deformity of the wrist is a congenital defect caused by a growth disturbance in the volar-ulnar distal radial physis leading to a typical appearance of the upper extremities. The majority of Madelung deformity case caused by hereditary dyschondrosteosis of the wrist. In a number of instances, the disease has a genetic etiology. This article reports the clinical and cytogenetic findings associated with Madelung deformity in 14 patients. Results indicate Madelung anomaly often is associated with additional clinical abnormalities, particularly delayed puberty and menstrual disorders, as well as sexual chromosome aberrations. PMID- 15790092 TI - The effect of total knee arthroplasty on body weight. AB - This prospective study quantified the weight change in 20 consecutive patients undergoing total knee arthroplasty. Resected bone, soft tissues, and bone reamings were collected during surgery and weighed using a digital scale at the end of the procedure. Results were compared to the cumulative weights of the prosthesis, bone cement, patellar component, and polyethylene liner. Average weight of the resected bone and soft tissues was 167.71 g for men and 130.13 g for women. Mean weight of the implanted prosthesis and cement used was 509.92 g for men and 422.56 g for women. Men tended to receive a larger-sized prosthesis than women. Overall, the average weight gain as a result of knee arthroplasty was 345.54 g for men and 292.44 g for women. This translates to an insignificant increase in body weight. PMID- 15790093 TI - Pyomyositis in adults without any predisposing factors in a non-tropical region. PMID- 15790094 TI - Management and identification of the etiological agent of a complicated case of necrotizing fasciitis. PMID- 15790095 TI - Multifocal osteoarticular tuberculosis. PMID- 15790096 TI - Bioconjugated nanoparticle for DNA protection from ultrasound damage. AB - To research whether poly-L-lysine-starch nanoparticle (PLL-StNP) could protect DNA from ultrasound damage or not, a series experiments were carried out: plasmid DNA-PLL-StNP complexes were treated with ultrasound for diverse times; the electrophoresis result proved that DNA bound to the complexes all the same. To investigate whether the conjugated DNA was completely protected or not, cDNA fragments bound to PLL-StNP were treated with ultrasound, and matched molecular beacons (MBs) were added. The cDNA-MB-PLL-StNP complexes exhibited dramatically increasing fluorescence, and had the same intensity as that of those MBs that were hybridized with free cDNA fragments. After being treated by ultrasound, the pIRGFP plasmid DNA-PLL-StNP complexes were transferred into COS-7 cells mediated by ultrasound. Green fluorescence protein expressed in most of the cells. Those results indicated that PLL-StNP could completely protect DNA from ultrasound damage. Furthermore, the DNA kept the same function as untreated one. PMID- 15790097 TI - Non-destructive rapid analysis discriminating between chromium(VI) and chromium(III) oxides in electrical and electronic equipment using Raman spectroscopy. AB - The European Union has banned chromium(VI) compounds in electrical and electronic equipment (EEE), such as chromate conversion coating films. Chromium(III) compounds are not banned. Using Raman spectroscopy without any preparation, we distinguished chromium(VI) oxide from chromium(III) oxide and chromium(III) hydroxide in chromate conversion coating films. Raman bands of chromium(VI) oxide were detected in films at around 1000 and 500 cm(-1), while chromium(III) compounds generated no bands in the region between 2000 and 200 cm(-1). The analysis took about 1 min, whereas the usual diphenylcarbazide-colorimetric method for analyzing chromium(VI) compounds takes about 10 h. PMID- 15790098 TI - Multielement determination of trace metals in river water (certified reference material, JSAC 0301-1) by high efficiency nebulization ICP-MS after 100-fold preconcentration with a chelating resin-packed minicolumn. AB - The determination of 34 trace metals in a river water certified reference material (CRM), i.e. JSAC 0301-1, which was issued by the Japan Society for Analytical Chemistry in January 2004, was performed by ICP-MS with a high efficiency nebulizer after preconcentration with a laboratory-made chelating resin-packed minicolumn, with which trace metals were concentrated 100-fold from 50 mL of a river water sample to 0.5 mL of the final analysis solution. Trace metals in JSAC 0301-1 were observed in the concentration range from 19 microg L( 1) of Al to 0.000053 microg L(-1) of Bi. It was found that most of the concentrations of trace metals, including rare earth elements (REEs), in JSAC 0301-1 were lower than those in JAC 0031, which was also a previously issued CRM prepared with water from the same river as that of JSAC 0301-1. The low concentrations of trace metals in JSAC 0301-1 might be attributed to the fact that there was a heavy rain before collecting the original water sample to prepare the present CRM. Furthermore, the REE distribution patterns of JSAC 0301 1, JAC 0031 and the average values of river water samples in Japan were parallel to each other. These results indicate that the distributions of REEs in JSAC 0301 1 and JAC 0031 were the typical ones of river water samples in Japan. PMID- 15790099 TI - Development of rapid plutonium analysis for environmental samples by isotope dilution/inductively coupled plasma mass spectrometry with on-line column. AB - This paper describes our development of a rapid on-line column/ID-ICP-MS technique for the analysis of plutonium (Pu) in environmental samples using an UTEVA extraction chromatograph resin (UTEVA resin) column. It took only 40 min to separate and measure Pu in the sample solution, including the time for conditioning the resin column for the next analysis. In our method, Pu in a 3 M nitric acid solution was fed to the UTEVA resin, and then eluted from the resin by reducing Pu to Pu(III) with 3 M nitric acid mixed with 0.01 M ascorbic acid after washing the resin. The outflow from the resin column was directly introduced to an ICP-MS system. The low concentration of ascorbic acid and the small volume of the eluting solution (0.6 mL) made successive stable analysis possible without any skimmer cone clogging. The chemical recovery of Pu during column operation was 70%, and typical lower detection limits for 239Pu, 240Pu and 242Pu were 9.2, 4.3 and 7.5 fg (21, 36 and 1.1 microBq), respectively. We analyzed five international standard materials for Pu, and obtained good results. PMID- 15790100 TI - Interpretation of seasonal variation of metals and abiotic properties in a tropical lake using multivariate analysis and MINTEQA2 program. AB - The hydrological cycle of Lake Ipe and the principal parameters that could affect its abiotic parameters and its metal (Fe, Mn, and Cd) concentrations were investigated. A detailed study of seasonal variations using Principal Components Analysis (PCA) for the most relevant metals (Fe, Mn and Cd) was performed, assisted by the MINTEQA2 geochemical equilibrium program using 15 water abiotic parameters. The abiotic variables and metals in the lake were governed by the hydrological regimen of the Parana River and by the occurrence of occasional flood pulses and rainfall. The occurrence of cadmium is a concern; its presence could be due to industrial or agricultural activities or even to a natural geochemical process. PMID- 15790101 TI - Interaction analysis of the carcinoembryonic antigen (CEA) with its monoclonal antibody immobilized on a gold surface using Fourier transform infrared reflection-absorption spectroscopy (FT-IR RAS). AB - A monoclonal antibody for the carcinoembryonic antigen (CEA) was immobilized on a gold chip surface covered by a self-assembled monolayer of 11-mercaptoundecanoic acid. Upon the addition of CEA, a Fourier transform infrared reflection absorption spectroscopy (FT-IR RAS) measurement showed an increased absorption at around 1500 - 1700 cm(-1), corresponding to its amide structures. Another addition of CEA polyclonal antibody on this chip caused a further increase of the absorption in this region only after a treatment with CEA. This result shows that an antibody-fixed gold surface coupled with an FT-IR RAS measurement provides a new tool for detecting the antibody-antigen interaction. PMID- 15790102 TI - Quantitative analysis of 17beta-estradiol in river water by fluorometric enzyme immunoassay using biotinylated estradiol. AB - A sensitive and simple immunoassay to determine 17beta-estradiol (E2) in fresh water was developed. The method is based on a solid-phase avidin-biotin binding assay and solid phase extraction. The binding event of E2 to the antibody is detected indirectly by the competitive reaction between E2 and biotinylated estradiol (BE) as a tracer for the limited binding sites of antibodies immobilized onto the wall of a microtiter plate. Namely, E2 concentrations are determined from the strong interaction between BE and avidin conjugated with horseradish peroxidase (avidin-HRP). In order to achieve a sensitive measurement for the binding of BE to the antibody immobilized on the microtiter plate substrate, QuantaBlu fluorogenic peroxidase substrate (QFPS) was employed. The detection limit and the linear range of E2 determination were 27 pM and 27 - 7480 pM, respectively. The relative standard deviations (RSD) for the E2 assay were between 0.3 and 12.0% (n = 3). The cross-reactivities of several other estrogens in this assay system were also investigated. No serious influences from any cross reaction caused by other estrogens tested in this experiment were observed. The determination of E2 in water samples from eight rivers and a marsh in Hokkaido was performed by the immunoassay combined with solid phase extraction. It was found that the concentration of E2 was in the range between 0.06 and 67 pM. PMID- 15790103 TI - Continuous in vivo monitoring of blood diffusible calcium using on-line microdialysis sampling coupled with flame atomic absorption spectrometry. AB - A direct, rapid and continuous in vivo monitoring of diffusible calcium in the blood of living rabbits has been developed using microdialysis sampling coupled on-line with flame atomic absorption spectrometry. Microdialysates perfused through implanted microdialysis probes were collected with a sample loop on an injection valve and directly introduced into the flame atomizer by a carrier solution. An ultrapure saline solution (0.9% NaCI, pH 7.2) was used as the perfusion solution at a flow rate of 20 microI min(-1) via the microdialysis probe. A 0.1% La solution in 0.5% HNO3 solution was employed as the carrier solution at a nebulizer uptake flow rate of 2.5 ml min(-1). The interval for each determination was 2.5 min (2 min of sampling time, 20 s of read time and 10 s of washing time). The performance characteristics of the on-line microdialysis-FAAS system were validated as follows: linearity range, 0 - 100 mg l(-1); detection limit (3a, n = 7), 3.66 mg l(-1); precision (RSD, n = 50), 6.2%. For the evaluation of analytical accuracy, the proposed on-line method was compared with the in vivo no net flux method. The use of an on-line microdialysis-FAAS system permitted the in situ, dynamic and continuous in vivo monitoring of diffusible calcium in the blood of the living rabbits after CaCl2 administration with a temporal resolution of 2.5 min. PMID- 15790104 TI - Determination of ATP as a fluorescence probe with europium(III)-doxycycline. AB - A new spectrofluorimetric method has been developed for the determination of adenosine disodium triphosphate (ATP). We studied the interactions between the doxycycline (DC)-Eu3+ complex and adenosine disodium triphosphate (ATP) by using UV-visible absorption and fluorescence spectra. Using doxycycline (DC)-Eu3+ as a fluorescence probe, under the optimum conditions, ATP could remarkably enhance the fluorescence intensity of the DC-Eu3+ complex at lambda = 612 nm. The enhanced fluorescence intensity of the Eu3+ ion was in proportion to the concentration of ATP. The optimum conditions for the determination of ATP were also investigated. The linear ranges for ATP were 1.00 x 10(-7) - 2.00 x 10(-6) mol L(-1) with detection limits of 4.07 x 10(-8) mol L(-1). This method is simple, practical and relatively free of interference from coexisting substances, and can be successfully applied to the determination of ATP in samples. The mechanism of fluorescence enhancement between the doxycycline (DC)-Eu3+ complex and ATP was also studied. PMID- 15790105 TI - Numerical taxonomy characterization of Baccharis genus species by ultraviolet visible spectrophotometry. AB - Numerical taxonomy characterization of Baccharis genus species was performed using ultraviolet-visible spectrophotometry. The aim was to present a more convenient, more practical, more economic and faster method based on chemometric methods and UV-vis absorbance to give the most information about species identity and discrimination, especially when their classification has been doubtful. Three Baccharis species: B. genistelloides Persoon var. trimera (Less.) DC, B. milleflora (Less.) DC, and B. articulata (Lam.) Persoon were included in the study. With the help of principal component analysis (PCA) and cluster analysis (CA), we could characterize the three species. Application of soft independent modeling of class analogy (SIMCA) and K-nearest neighbor (KNN) methods on a training set of 65 extracts resulted in models that correctly classified all samples of an independent validation set, eight samples of B. genistelloides Persoon var. trimera (Less.) DC and one sample donated by Prof. Alarich Schultz Herbarium, Porto Alegre-RS, Brazil. PMID- 15790106 TI - Analysis of tetracycline antibiotics using HPLC with pulsed amperometric detection. AB - The analysis of tetracycline, oxytetracyline, chlortetracycline and doxycycline by high-performance liquid chromatography with pulsed amperometric detection using an anodized boron-doped diamond thin film (BDD) electrode is originally reported. The analyses were carried out using the mobile phase, phosphate buffer (0.01 M, pH 2.5)-acetonitrile (80:20; v/v), on a C18 column (250 x 4.6 mm i.d., 5 microm) at a flow rate of 1.0 mL/min. The optimal PAD waveform parameters at the anodized BDD were 1.5 V (versus Ag/AgCl) detection potential (E(det)) for 290 ms (200 ms delay time and 90 ms integration time), 2.0 V (versus Ag/AgCl) oxidation potential (E(oxd)) for 200 ms oxidation time (t(oxd)) and 0.4 V (versus Ag/AgCI) reduction potential (E(red)) for 200 ms reduction time (t(red)). The proposed method showed the simultaneous determination of tetracycline, oxytetracyline, chlortetracycline and doxycycline with a linear range of 0.1 - 100 microg/mL, detection limits of 0.05 - 0.1 microg/mL and recoveries of 70.8 - 96.0%. The application of this method to real samples was demonstrated and validated using a shrimp sample. PMID- 15790107 TI - Rapid determination of paeoniflorin and three sugars in Radix Paeoniae Alba by capillary electrophoresis. AB - A method based on capillary electrophoresis with electrochemical detection has been developed for the separation and determination of paeoniflorin, sucrose, glucose, and fructose in traditional Chinese medicine, Radix Paeoniae Alba. The effects of several important factors, such as the concentration of NaOH, the separation voltage, the injection time, and the detection potential, were investigated to determine the optimum conditions. The detection electrode was a 300-microm diameter copper disc electrode at a working potential of +0.60 V (versus SCE). The four analytes can be well separated within 8 min in a 40 cm length fused-silica capillary at a separation voltage of 12 kV in a 75 mM NaOH aqueous solution. The relation between the peak current and the analyte concentration was linear over about 3 orders of magnitude with detection limits (S/N = 3) ranging from 1 to 2 microM for all analytes. The proposed method has been successfully applied for the determination of the paeoniflorin and sugars in real plant samples with satisfactory assay results. PMID- 15790108 TI - Chiral speciation and determination of DL-selenomethionine enantiomers on a novel chiral ligand-exchange stationary phase. AB - A new type of chiral ligand-exchange stationary phase (CLES) was successfully synthesized by treating silica gel with beta-(3,4-epoxycyclohexyl)ethyltrimethoxy silane and opening the epoxy ring by L-isoleucine. The chiral speciation of DL selenomethionine (DL-SeMet) by high-performance liquid chromatography (HPLC) with UV absorbance on the CLES column was studied. The influences of the contents of copper ion and methanol as well as the pH value in the mobile phase and temperature of the column on the efficiency of resolution of DL-SeMet were investigated in detail. DL-SeMet could be completely resolved within 40 min under the optimal operating conditions of 0.1 mmol/L Cu2+ at 0.05 mol/L KH2PO4 buffer (pH = 5.5) and 35 degrees C temperature of the column. The limits of detection of D- and L-SeMet were 255 ppb and 286 ppb, respectively. This method was applied to determine the D- and L-enantiomers of DL-SeMet in real samples, such as selenized yeast powder and garlic. PMID- 15790109 TI - Separation and determination of trace amounts of aluminium(III), vanadium(V), iron(III), copper(II) and nickel(II) with CALKS and PAR by RP-HPLC. AB - An RP-HPLC method for the separation and determination of aluminium(III), vanadium(V), iron(III), copper(II) and nickel(II) with CALKS (Chromazol KS) and PAR ([4-(2-pyridylazo)resorcinol]) chelating on a YWG-ODS column was developed. A mixture of methanol-tetrahydrofuran(THF)-water (60:5:35 v/v) containing 0.2 mol/L LiCl, 5 x 10(-5) mol/L CALKS, 5 x 10(-5) mol/L PAR and acetate buffer solution (pH 4.9) was selected as mobile phase. The method has high sensitivity, with the detection limits being 6 ng/mL for aluminium(III), 3.5 ng/mL for vanadium(V), 10.4 ng/mL for iron(III), 6.3 ng/mL for copper(II) and 8.7 ng/mL for nickel(II). It also has good selectivity, so that most foreign metal ions do not interfere under the optimum conditions. The method can be applied to the simultaneous determination of trace amounts of aluminium, vanadium, iron, copper and nickel in rice and flour samples. PMID- 15790110 TI - Trace and ultratrace analysis of purified water samples and hydrogen peroxide solutions for phosphorus by flow-injection method. AB - A highly sensitive fluorescence-quenching method for the determination of phosphorus based on the formation of an ion associate between molybdophosphate and Rhodamine B (RB) was developed. A simple flow-injection system coupled with a fluorescence detector was used to measure the fluorescence intensity at 560 nm and 580 nm as an excitation and an emission wavelength, respectively. The calibration graph for phosphorus showed a good linearity in the range of (0 - 1) x 10(-7) M (1 M = 1 mol L(-1)), and a detection limit of 1 x 10(-9) M (S/N = 3). The proposed method was successfully applied to the determination of ultratrace amounts of phosphorus in ultrapurified and purified water samples, and to the determination of trace amounts of phosphorus in commercially-available hydrogen peroxide solutions with satisfactory results. PMID- 15790111 TI - Extraction mechanism for palladium(II) from hydrochloric acid solution with 2 dodecylthiomethylpyridine using a stirred transfer cell. AB - 2-Dodecylthiomethylpyridine (DTP) was newly synthesized to study its extraction properties for precious metals. DTP was a selective extractant for palladium(II) and gold(III) over base metals. The loading test for palladium(II) showed that one palladium ion reacted with one molecule of DTP. The extraction rate of palladium with DTP was measured using a Lewis-type transfer cell at 303 K. The extraction reaction of palladium with DTP has been found to be a first order reaction with respect to palladium ion, DTP, and hydrogen ion concentrations. This reaction is inversely proportional to chloride ion concentration. The rate determining step was the parallel reactions of DTP with PdCl3(-) and PdCl4(2-) in the aqueous phase. PMID- 15790112 TI - Determination of trace sodium in the water-steam system of power plants using an FIA/ISE method with an automatic penetration and alkalization apparatus. AB - A new method for the rapid determination of trace-level sodium ion based on flow injection analysis (FIA) and an ion selective electrode (ISE) is proposed. Various effects on the sensitivity of the method such as the flow rate and alkalinity of the carrier, sampling volume, temperature, length of reaction coil, length and thickness of alkalization tube, concentration of the alkalizing reagent etc. were investigated. The optimum conditions were ascertained. The method showed good linearity in the concentration ranges of 0.5 - 10 microg L(-1) and 10 - 100 microg L(-1), and could deal with 40 - 50 samples per hour. The consumption of the sample is only 0.80 mL per time. The relative standard deviation was 0.55%, and the recovery range was 98 - 103%. By designing a hermetically sealed single-line FIA-ISE manifold, a problem removing the interference of sodium ion from air could be solved, and automatic alkalization of the sample was realized. This method has been used for successfully determining the trace-level sodium ions in the water-steam system at fossil power plants. PMID- 15790113 TI - Adsorption behavior of metal ions onto a bovine serum albumin (BSA) membrane monitored by means of an electrode-separated piezoelectric quartz crystal. AB - The interaction between metal ions and bovine serum albumin (BSA) was studied by using a piezoelectric quartz crystal (PQC) arranged in the electrode-separated configuration. A silanized surface of the PQC was coated with a BSA membrane via a coupling reaction with glutaraldehyde. The frequency shifts obtained from PQC coated with a BSA membrane suggested that various kinds of metal ions could be adsorbed onto the BSA membrane from aqueous solutions containing a low concentration of metal ions (2 or 10 micromol dm(-3)), only when the BSA was denatured with an alkaline solution. Anionic species of Pt(IV) and Au(III) were adsorbed onto the denatured BSA membrane from an acetic acid solution at pH 2.2, and cationic species of Cd(II), Zn(II), Co(II), Ni(II), Cu(II), and Ag(I), and cations, such as Ca2+, Ba2+, and Mg2+, were adsorbed from ammonia buffer at pH 9.5, whereas Al(III), Cr(III), Fe(III), Hg(II), and Pb(II) were hardly adsorbed. The adsorption mechanisms of these metal ions are discussed, based on the electrostatic interaction between the metal ions and the denatured BSA membrane, and complex formation between the metal ions and amino acid residues of the denatured BSA. Further, the PQC coated with a denatured BSA membrane was applied to the determination of Pt and Cd, using large frequency shifts for Pt(IV) and Cd(II). PMID- 15790114 TI - Potentiometric membrane electrode for salicylate based on an organotin complex with a salicylal Schiff base of amino acid. AB - A novel salicylate-selective electrode based on an organotin complex with a salicylal Schiff base of amino acid salicylaldehydeaminoacid-di-n-butyl-Sn(IV) [Sn(IV)-SAADB] as ionophore is described, which exhibits high selectivity for salicylate over many other common anions with an anti-Hofmeister selectivity sequence: Sal- >> PhCOO- > SCN- > Cl04- > I- > NO3- > NO2- > Br- > Cl- > CH3COO-. The electrode, based on Sn(IV)-SAADB, with a 30.44 wt% PVC, a 65.45 wt% plasticizer (dioctyl phthalate, DOP), a 3.81 wt% ionophore and a 0.3 wt% anionic additive is linear in 6.0 x 10(-6) - 1.0 x 10(-1) mol l(-1) with a detection limit of 2.0 x 10(-6) mol l(-1) and a slope of 62.0 +/- 1.2 mV/decade of salicylate concentration in a phosphate buffer solution of pH 5.5 at 25 degrees C. The influence on the electrode performances by lipophilic charged additives was studied, and the possible response mechanism was investigated by UV spectra. The electrode was applied to medicine analysis and the result obtained has been satisfactory. PMID- 15790116 TI - Charge-transfer triiodide ion-selective electrode based on 7,16-dibenzyl 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane. AB - A new PVC membrane electrode for the triiodide ion based on a charge-transfer complex of iodine with 7,16-dibenzyl-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane as a membrane carrier was prepared. The electrode exhibits a Nernstian response for triiodide ions over a wide concentration range (1.0 x 10(-1)-1.0 x 10(-5) M) with a slope of 59.3 +/- 0.9 mV decade(-1) and a detection limit of 6.3 x 10(-6) M. It has a response time of 30 s and can be used for at least 3 months without any divergence in the potential. The potentiometric response is independent of the pH, in the pH range 1.6 - 10.0. The proposed electrode has shown very high selectivity for the triiodide ion over a wide variety of other anions. This electrode was successfully applied as an indicator electrode in the potentiometric titration of ascorbic acid and hydroquinone from pharmaceutical preparations as well as ascorbic acid in orange juice and dissolved O2 in tap water. PMID- 15790115 TI - Strontium(II) sensor based on a modified calix[6]arene in PVC matrix. AB - 5,11,17,23,29,35-Hexakis(1,1,3,3-tetramethylbutyl)-37,38,39,40,41,42 hexakis(carboxy methoxy)calix[6]arene (I) has been evaluated as an ionophore for the analysis of Sr2+. The influences of the nature of the plasticizers (DBA, CN, DOP, NPOE) and of the anion excluder (NaTPB) on the characteristics of the electrode were discussed. The best electrode was fabricated with a membrane having composition 6:150:170:3 (I:PVC:DBA:NaTPB). The response to Sr2+ was Nernstian in the range 1.9 x 10(-5) to 1.0 x 10(-1) M of Sr2+. The influence of pH has also been studied. The electrode exhibited better potential stability and had an operational lifetime of 4 months. The K(A,B)(Pot) values showed that other alkaline earth metal ions are well discriminated. The sensor has also been used as an indicator electrode in the potentiometric titration of sodium carbonate with strontium(II) ions. PMID- 15790117 TI - Molecular probe for a fluorous medium: long-lived phosphorescence of alpha diketones in perfluoromethylcyclohexane at room temperature. AB - We found that alpha-diketones (2,3-butanedione (BD) and 1-phenyl-1,2-propanedione (PPD)) were very suitable luminescence probes for studying the properties of a perfluorinated solvent (i.e., fluorous solvent; perfluoromethylcyclohexane (PFMC)), since these compounds were soluble in PFMC and showed long-lived phosphorescence even at room temperature. The phosphorescence lifetime (tau(p)) of BD in PFMC (650 micros) was much longer than that in cyclohexane (CH, 270 micros). The longer tau(p) value of BD in PFMC was ascribed to the variation of the intersystem crossing rate constant (k(isc)) from the excited triplet state (T1) to the ground state (S0) with the solvent. Some possible reasons for the change in k(isc) were discussed in terms of solute-solvent interactions. Furthermore, by utilizing phosphorescence quenching of BD by pyrene, we, determined a rate constant of the diffusion-controlled reaction in PFMC. Characteristic behaviors of mixing/separation processes between PFMC and a common organic solvent observed by Schlieren photographs were also reported. PMID- 15790118 TI - Electronic structure analysis of iron(III)-porphyrin complexes by X-ray absorption spectra at the C, N and Fe K-edges. AB - X-ray absorption near edge structure (XANES) measurements at the C, N, and Fe K absorption edges were performed for iron(III)-tetraphenylporphyrin (FeTPP), iron(III)-tetrakis(p-carboxyphenyl)porphyrin (FeTCPP), and iron(III)-tetrakis(p sulfonatophenyl)porphyrin (FeTSPP). The spectral shapes differ in the Fe K XANES, but not in C and N K XANES among FeTPP, FeTCPP, and FeTSPP. Crosschecks of XANES data for C, N, and Fe K absorption edges in combination with discrete variational (DV)-Xalpha molecular orbital (MO) calculations indicate that each p-electron withdrawing group on four meso-phenyl substitutes in an Fe(III)-porphyrin complex brings about a unique electron state through the complex because of the electron withdrawal strength, itself. Consequently, they affect the positive charge of the center Fe(III) ion. PMID- 15790119 TI - Pulsed NMR study of network formation in the course of bulk polymerization of methyl acrylate. AB - The proton spin-spin relaxation time (T2) during the bulk polymerization of methyl acrylate was measured as a function of the reaction time at various temperatures. Three kinds of T2 (T2L (long), T2S (short) and T2M (intermediate)) were obtained as the polymerization proceeded. The fraction of T2S (FS) increased sigmoidally at a certain reaction time, while that of T2L (FL) decreased reciprocally. The former corresponded to the amount of a polymer whose molecular weight was sufficiently high enough to cause a tight entanglement that produced a transient network structure; the latter reflected a decrease in the mixture of the monomer and the low molecular weight of the polymer. T2M is considered to arise from a relatively mobile region of the entanglement. The relationship between the fractions of T2S + T2M and the polymer yield was found to be linear, which led us to monitor the polymer yield in real time during the polymerization in a non-distractive manner. 13C DD (dipolar decoupling)/MAS (magic angle spinning) NMR spectra were also measured to monitor the polymerization process in terms of the molecular motions between the main chain and the side chain in the formation of a network structure. The 13C DD/MAS NMR spectra show that the side chain motion became restricted as well as the main chain when the "Trommosdorff effect" (gel effect) was observed, and a part of the monomers were trapped in the network structure. PMID- 15790120 TI - Characterization of zeolite NaA membrane by FTIR-ATR and its application to the rapid evaluation of dehydration performance. AB - A zeolite NaA (LTA) membrane supported by an alumina porous support tube was characterized by Fourier Transform Infrared Attenuated Total Reflectance method (FTIR-ATR) with a diamond prism as the waveguide. A method using the FTIR-ATR was developed to estimate rapidly the EtOH/H2O pervaporation (PV) performance of the membrane. The Si-O asymmetric stretching vibration region of LTA membrane spectra synthesized hydrothermally on seeded alumina substrates showed a bimodal peak (830 - 1200 cm(-1)). The two peaks were assigned to a surface LTA directly derived from the seed crystal (1012 cm(-1)), and to LTA and/or amorphous substances embedded in the alumina porous support (930 cm(-1)). The spectrum from LTA membrane synthesized on nonseeded alumina substrate, however, showed a single broad peak similar to the powder-formed one. These results indicate that the Si-O spectral shape of the LTA membrane is influenced strongly by the synthesis method. Also, the FTIR-ATR of the LTA membrane can detect the Si-O peaks as part of the depth information. It was first shown that the relative ratio (930 cm( 1)/1012 cm(-1)) of the two Si-O peaks from the LTA membranes on seeded alumina substrates closely relates to the water selectivity (alpha) in the PV of EtOH/H2O mixture; the alpha increases exponentially with the peak ratio. This result suggests that the differences in the vertical distribution of LTA crystal and amorphous material strongly affect the dehydration performance in the EtOH/H2O PV, that is, the amorphous-like material embedded in the alumina porous support plays an important role. The relative peak ratio measurement can be used for the rapid evaluation of the dehydration performance of the membrane. PMID- 15790121 TI - Multivariate calibration analysis for metal porphyrin mixtures by an ant colony algorithm. AB - A new biomimetic algorithm, Chemical Ant Colony Algorithm, has been developed, which has the characteristics of intelligent search, global optimization, robustness, distributed computation and easy combination with other heuristic. The proposed method has been successfully applied to the spectroscopy analysis of the Zn2+, Cd2+, Pb(2+)-porphyin tribasic color system with supramolecular properties; the errors are within +/-8.0%. PMID- 15790122 TI - Analysis of ice water by the thermally stimulated depolarized current (TSDC) method. AB - The thermally stimulated depolarized current and temperature profiles (TSDC analysis) on iced water was proved to be an effective tool for the qualitative evaluation of various water samples, each having an independent relaxation process, brought about by a pure dipolar orientation. The method proved to be applicable to different kinds of water samples used in the experiment and tap water samples from different suppliers. Five main peaks (A to E, in the order of increasing temperature) were observed, of which two peaks (B and D) were found for the first time. The appearance of multiple peaks suggested the existence of multi-states of the hydrogen bond, cleaved by the TSDC process. The TSDC profiles were quite reproducible when the water samples contained practically no cations. A separate addition of each cation at a low concentration level revealed that a cation with a smaller ionic radius shifted peak A to a higher temperature. For ice of tap water, which contained relatively higher amount of cations, the TSDC profiles were quite different in shape compared with the standard ice-water samples (shift of peaks A - D to higher temperature, and a strong increase in the current strength of peaks B - E). However, it was still possible to tell from which districts the water samples were supplied. PMID- 15790124 TI - The liquid-liquid extraction using hydrophilic solvents occluded to an oil adsorbent as an organic phase: extraction of 8-quinolinolato complexes of alkaline earth metals into tetrahydrofuran. AB - In order to use a hydrophilic solvent as an organic phase in liquid-liquid extraction by retaining it to a solid support, the occlusion and desorption of nine hydrophilic solvents, such as methanol, to three kinds of commercially available oil-adsorbent were examined. The combination of tetrahydrofuran and WOSEP made by fibrous polypropylene was finally selected, and was subjected to the extraction of alkaline Earth metals with 8-quinolinol. It was proved that Mg2+, Ca2+, and Sr2+ ions could be successfully extracted into the tetrahydrofuran phase in the form of 8-quinolinolato complexes, which could not be extracted into CHCl3. PMID- 15790123 TI - Determination of oxycodone and hydrocotarnine in cancer patient serum by high performance liquid chromatography with electrochemical detection. AB - We developed an HPLC procedure using electrochemical detection for the quantitation of oxycodone and hydrocotarnine in cancer patients serum. An eluent of methanol:acetonitrile:5 mM pH 8 phosphate buffer (2:1:7) was used for the mobile phase. The calibration curve was linear in the range from 10 ng/mL to 100 ng/mL. The recovery of oxycodone and hydrocotarnine was 97.2% and 90.5%, respectively. The relative standard deviations within-runs and between-runs for the assay of oxycodone or hydrocotarnine were less than 4.8%. The method developed here was better than the method reported previously. PMID- 15790125 TI - Equivalence of the acute cytokine surge and myocardial injury after coronary artery bypass grafting with and without a novel extracorporeal circulation system. AB - Cardiopulmonary bypass (CPB) contributes to a morbidity-inducing systemic inflammatory response after cardiac surgery. We compared this response in patients receiving coronary artery bypass grafting (CABG) with (CPB group; n = 7) or without (off-pump group; n = 8) the Minimal Extracorporeal Circulation (MECC) system. Serum concentrations of tumour necrosis factor (TNF)-alpha, soluble TNF receptors, pro- and anti-inflammatory interleukins (ILs) and other myocardial injury markers were measured after anaesthetic induction, at 1 h, 4 h and 24 h after completing all anastomoses or serially. Soluble TNF receptor type I (sTNFRI) and IL-8 peaked early after CABG in both groups and did not decline. Serum sTNFRI was significantly higher in the CPB compared with the off-pump group at 1 h, whereas IL-8 was significantly lower in the CPB group throughout. The MECC system, therefore, produces an equivalent acute cytokine response and degree of myocardial injury to off-pump CABG, and may be useful when CABG cannot be performed without CPB. PMID- 15790127 TI - Examination of visceral perception and gastric tone by gastric stimulation using air inflation during endoscopy. AB - This study investigated the assessment of visceral perception and gastric tone using insufflation during endoscopy. The intragastric pressure was measured during insufflation in 16 patients with gastro-oesophageal reflux disease, 25 patients with functional dyspepsia and 24 normal controls, using a pressure transducer inserted through the forceps aperture until gastric symptoms appeared. The intragastric pressure at the visceral perception threshold, the time to threshold and the time-pressure curve gradient were measured. The visceral perception threshold was significantly reduced in the functional dyspepsia group compared with the normal controls. Balloons were also inserted into the duodenal bulbs of 10 normal controls to investigate the effect of efflux of air into the duodenum; the measured parameters were not influenced by inflation and deflation. These results suggest that it is possible to assess visceral perception and gastric tone by measuring changes in intragastric pressure, using gastric stimulation by insufflation during endoscopy. PMID- 15790126 TI - Effect of angiotensin-converting enzyme inhibitors on phenylephrine responsiveness in patients with valvular heart disease. AB - We studied patients with valvular heart disease to investigate whether chronic preoperative treatment with angiotensin-converting enzyme (ACE) inhibitors modulates the effect of phenylephrine (PE) on anaesthesia-induced hypotension. Sixty-five patients were enrolled in the study and hypotension developed after anaesthesia in 36 (18 in the control group and 18 in the ACE inhibitor group). These patients received PE infusions, which were increased in a stepwise fashion at 10-min intervals. Increased mean arterial pressure due to PE infusion was significant only in the control group. There was no significant difference in pressor response or change in haemodynamic variables with PE infusion between the two groups. Treatment with ACE inhibitors did not increase the incidence of hypotensive episodes or significantly modify pressor response after anaesthesia in patients with valvular heart disease. PMID- 15790128 TI - Effect of antihypertensive therapy on blood rheology in patients with essential hypertension. AB - Hypertension is an important risk factor for cardiovascular disease, and antihypertensive drugs can decrease the occurrence of such events in hypertensive patients. This study compared the rheological properties of blood in 22 untreated hypertensive patients, 42 patients taking antihypertensive drugs and 74 normotensive subjects. Using a microchannel method, the whole blood passage time was measured and blood movement was observed with a microscope connected to an image display unit. The blood passage time in untreated hypertensive patients was significantly higher than in treated hypertensive patients or normotensive subjects, but was similar in the latter two groups. Microscopic observations showed that platelet aggregation and leucocyte adhesion were increased in untreated hypertensive patients, resulting in poor flow, while blood samples from treated hypertensive patients and normotensive subjects passed smoothly through the microchannels. These rheological differences could contribute to the decrease in cardiovascular disease seen when hypertensive patients are treated effectively. PMID- 15790129 TI - Evaluation of the antimicrobial activity and materials compatibility of orthophthalaldehyde as a high-level disinfectant. AB - We tested the antimicrobial activity of orthophthalaldehyde (OPA) against 21 strains (16 species) of pathogenic microorganisms that cause hospital-associated infections. Changes in hepatitis B surface antigen (HBs-Ag) resulting from the addition of OPA to HBs-Ag-positive serum were measured using a radioimmunoassay. We also examined the effect of immersing medical instruments in OPA (0.55%) for 168 h at room temperature. OPA (0.5%, 0.37% and 0.25%) killed 11 strains of vegetative bacteria within 15 s, and it killed the test micro-organisms faster than 3.0% glutaraldehyde (GTA). Incubation with OPA or GTA caused levels of HBs Ag to fall below a cut-off value within 30 s. OPA did not adversely affect instruments made from various materials. OPA demonstrated more effective antimicrobial activity than GTA against a range of microorganisms. We conclude that OPA should replace GTA as the first-choice high-level disinfectant for endoscopes, considering its antimicrobial efficacy and low inhalation toxicity. PMID- 15790130 TI - Pre-emptive effects of ibuprofen syrup and lidocaine infiltration on post operative analgesia in children undergoing adenotonsillectomy. AB - We aimed to compare the effects of lidocaine and adrenaline with ibuprofen syrup (administered before adenotonsillectomy) on post-operative analgesia and initiation of oral feeding. One group of 20 children received 100 g/5 ml ibuprofen suspension (10 mg/kg) 1 h before anaesthesia; bleeding control was provided by pre-incisional administration of 1:200,000 adrenaline solution (10 ml). The same amount of 0.5% lidocaine solution plus 1:200,000 adrenaline was applied pre-incisionally in a similar manner in a second group of 20 children. No significant differences were observed between the two groups in terms of the duration of operation and anaesthesia, post-operative pain scores, paracetamol requirements, times to initiation of liquid and solid food intake, or adverse side-effects. We conclude that ibuprofen syrup applied pre-incisionally and local infiltration with lidocaine are equally effective for post-operative analgesia. PMID- 15790131 TI - Effects of N-acetylcysteine treatment on oxidative stress in acetic acid-induced experimental colitis in rats. AB - We assessed the possible protective effects of N-acetylcysteine (NAC) against toxic damage in the rat colon. Two doses of NAC (20 mg/kg and 100 mg/kg) given for 2 days and 7 days after acetic acid administration (to induce colitis) were tested. NAC was dissolved in saline and administered locally (intracolonic), systemically (intraperitoneal) or in a combination (intracolonic and intraperitoneal). Several parameters, including macroscopic and histopathological scores and myeloperoxidase, glutathione and nitric oxide concentrations were measured using standard assay procedures. Treatment with 100 mg/kg NAC for 7 days significantly decreased tissue myeloperoxidase, glutathione and nitric oxide concentrations. The 20 mg/kg dose had no protective effects. The data indicate that NAC substantially reduced the degree of colonic injury, probably by regulating free radical production and inhibiting inflammation. It may, therefore, have a role in the treatment of inflammatory bowel disease. PMID- 15790132 TI - Adiponectin replacement therapy attenuates myocardial damage in leptin-deficient mice with viral myocarditis. AB - The effects of adiponectin replacement therapy on myocardial damage were studied in leptin-deficient (OB) mice with acute viral myocarditis. Encephalomyocarditis virus was injected intraperitoneally into OB and wild-type (WT) mice. One subgroup of OB mice received no intervention and another subgroup received daily adiponectin replacement, simultaneously with viral inoculation. Differences in heart weight, cardiac histological score, numbers of infiltrating or apoptotic cells in the myocardium and the immunoreactivity of adiponectin receptors in myocytes were determined. The reactivity of adiponectin receptor 1 in myocytes from OB mice on day 4 and day 8 after viral inoculation was significantly decreased compared with that in myocytes from WT mice; the OB mice also had elevated cardiac weights and severe inflammatory myocardial damage. Adiponectin replacement in OB mice inhibited the development of severe myocarditis by augmenting myocyte adiponectin receptor 1 reactivity. Exogenously administered adiponectin may inhibit the progression of viral myocarditis through binding to the adiponectin receptor 1 in leptin-deficient conditions. PMID- 15790133 TI - The evaluation of 13 patients with intrathoracic extrapulmonary hydatidosis. AB - Cases of intrathoracic extrapulmonary hydatid cysts are very rare. We identified 13 patients with intrathoracic extrapulmonary hydatid cysts in our clinic over 12 years. Four patients had extrapulmonary cysts only; nine patients had both intrapulmonary and extrapulmonary cysts. Cysts were identified in the pleural space, extrapleural region, diaphragm and chest wall. Thoracotomy was used in all patients, and extrapulmonary lesions were removed by cyst extirpation from surrounding tissue or by pericystectomy. In one patient with chest wall involvement, partial rib resections were performed because of rib destruction. In two patients with liver cysts passing through the diaphragm to the thorax, the diaphragm was cut, cysts on the liver roof were removed and then the diaphragm was repaired. There was no mortality, morbidity, or disease recurrence during the post-operative period in any of the 13 patients. We conclude that these rare cases give a new insight into hydatid cyst pathophysiology. PMID- 15790134 TI - Gastric carcinoma and thyroid status. AB - Gastric carcinoma is reported to be more frequent in geographical areas where diets are either iodine-deficient or iodine-excessive. Reports have also shown an association between thyroid diseases and some of the risk factors for gastric carcinoma. We investigated the frequency of thyroid disorders in 61 patients with gastric carcinoma compared with 55 healthy control subjects. Thyroid health was evaluated by physical examination and by measuring the serum levels of thyroid hormones and thyroid autoantibodies. More patients with gastric cancer had goitre compared with healthy controls (49.1% versus 20%, respectively). Significantly more patients with gastric cancer had non-toxic goitre compared with control subjects. There was also a significant difference in the incidence of autoimmune thyroid disease--27.8% of patients with gastric cancer versus 10.9% of control subjects were affected. These results indicate that there is a significant association between gastric cancer and thyroid disorders. PMID- 15790135 TI - Prognostic significance of thrombocytosis in node-negative colon cancer. AB - Several clinical studies have shown that thrombocytosis is a poor prognostic factor in some types of cancer, but data about the impact of thrombocytosis on prognosis in patients with colon cancer are very limited. We investigated the prevalence and prognostic effect of pre-operative thrombocytosis, defined as a platelet count > 400 x 10(9)/l, retrospectively in patients with node-negative colon cancer. Out of 198 patients, 24 (12.1%) had thrombocytosis, and its presence correlated with tumour depth and lymphatic invasion. Univariate analysis revealed that disease-free survival and overall survival were shorter in patients with pre-operative thrombocytosis than those without thrombocytosis. On multivariate analysis, thrombocytosis alone retained significance as a poor prognostic factor for both disease-free survival and overall survival. In conclusion, this study shows an association between thrombocytosis and poor survival in patients with node-negative colon cancer. The preoperative platelet count may help to identify patients with an unfavourable prognosis in this subgroup. PMID- 15790136 TI - Adiponectin, T-cadherin and tumour necrosis factor-alpha in damaged cardiomyocytes from autopsy specimens. AB - This study determined the presence of adiponectin, T-cadherin (an adiponectin receptor) and tumour necrosis factor-alpha (TNF-alpha) in damaged myocytes from autopsied patients with acute or old myocardial infarction (MI) or dilated cardiomyopathy (DCM), using immunohistochemical staining. The enrolled patients included eight with acute MI, six with old MI and seven with DCM. Four autopsied individuals with no cardiac lesions were also enrolled as controls. Adiponectin and TNF-alpha were not observed in normal myocytes from control subjects, but T cadherin was weakly detected. Immunoreactivity for adiponectin and T-cadherin was observed at the periphery of damaged myocytes from MI and DCM patients; intracellular reactivity for TNF-alpha was also seen. There were no statistically significant differences in the degree of reactivity for each molecule in the myocytes between the MI and DCM patients. These results suggest that the presence of adiponectin and TNF-alpha in damaged myocytes may contribute to the processes of myocardial injury occurring in MI and DCM. PMID- 15790137 TI - Solid gastric emptying after highly selective vagotomy and pyloroplasty in patients with obstructing duodenal ulcer. AB - Before being superseded by medical management, highly selective vagotomy (HSV) without drainage was the procedure of choice for uncomplicated duodenal ulcer. It is also justified for complications, including perforation and bleeding in selected cases. This prospective study evaluated the effects of HSV plus drainage on solid gastric emptying in 20 patients with chronic duodenal ulcer and pyloric stenosis. Patients were treated with HSV plus pyloroplasty (Heineke-Mikulicz pyloroplasty in five patients, Finney pyloroplasty in six patients and Jaboulay gastroduodenostomy in nine patients) and underwent solid-phase gastric emptying scintigraphic studies pre-operatively and 2 months and 6 months post-operatively. Results were compared with those from 10 controls. No significant differences were observed between the different types of pyloroplasty, although emptying was slightly faster in the gastroduodenostomy group. Gastric emptying returned to normal by 6 months post-operatively. In conclusion, HSV plus pyloroplasty is effective and can be used for the relief of stenosis in selected cases of duodenal ulcer. PMID- 15790138 TI - Reliability and validity of the Modified Dental Anxiety Scale in Turkish patients. AB - We aimed to determine dental anxiety among Turkish patients and assess the sensitivity, specificity, positive/negative predictive values and reliability of the Modified Dental Anxiety Scale (MDAS) and Corah's Dental Anxiety Scale (DAS). Patients referred to our clinic for dental treatment who had a history of dental anxiety were included in the study. 294 randomly selected patients (mean age 38.8 years) completed a questionnaire combining Corah's DAS and MDAS. They were retested 15 days later. The prevalence of dental anxiety was found to be 9.9% (29/294) for Corah's DAS at the cut-off point > or = 15 and 8.8% (26/294) for the MDAS at the cut-off point > or = 19. Both dental anxiety scales gave acceptable sensitivity, specificity, positive and negative predictive values at these cut off points. PMID- 15790139 TI - Usefulness of induced sputum and fibreoptic bronchoscopy specimens in the diagnosis of pulmonary tuberculosis. AB - We investigated the diagnostic value of induced sputum (IS) and bronchial lavage (BL) specimens in patients with suspected pulmonary tuberculosis who had negative spontaneous sputum specimens or who were unable to produce sputum spontaneously. IS specimens and BL specimens obtained using flexible fibreoptic bronchoscopy from 55 patients were evaluated for the presence of acid-fast bacilli (AFB) and cultured for Mycobacterium tuberculosis. Positive results were found with IS smear in 23 patients, BL smear in 26 patients, and IS or BL culture in 42 patients. Culture of BL specimens had a higher sensitivity than IS or BL smears or culture of IS specimens. The highest sensitivity rate was obtained with a positive BL or IS culture (86%). For early diagnosis (a positive IS or BL smear), the sensitivity was 57%. IS has a higher sensitivity rate than spontaneous sputum for the detection of tuberculosis, and fibreoptic bronchoscopy is useful for the early diagnosis of tuberculosis when AFB are not detected in spontaneous or induced sputum specimens. PMID- 15790140 TI - Epidemiology of oral contraceptive related thrombosis. PMID- 15790141 TI - Hemostasis in normal pregnancy. PMID- 15790142 TI - Inherited platelet abnormalities. PMID- 15790143 TI - Inherited thrombophilia and pregnancy loss-epidemiology. PMID- 15790144 TI - Prothrombotic mechanisms in preeclampsia. PMID- 15790145 TI - Mechanisms of thrombosis in cancer. PMID- 15790146 TI - Infertility and thrombophilia. PMID- 15790147 TI - Antiphospholipid syndrome: clinical manifestations and management. PMID- 15790148 TI - Antithrombotic prophylaxis for pregnancy loss. PMID- 15790149 TI - Management of valvular heart disease. PMID- 15790150 TI - The protein C system (TM EPCR). PMID- 15790151 TI - Immune thrombocytopenia in the fetus and neonate: diagnosis and therapy. PMID- 15790152 TI - Bleeding disorders in neonates. PMID- 15790153 TI - Counseling of women with thrombophilia. PMID- 15790154 TI - Management of thrombophilia in women with PVC. PMID- 15790155 TI - Prevention and treatment of VTE in women with cancer. PMID- 15790156 TI - Pathogenesis and treatment of DIC. PMID- 15790157 TI - Development of hemostasis in the fetus. AB - The fetal hemostatic system is unique in many respects. Many coagulation proteins are expressed early in embryonic development and play roles outside of coagulation in cell proliferation and differentiation. Coagulation mRNA and protein can be detected within hepatic and endothelial cells within the first month of gestation and in fetal plasma during the third month. However, development of levels of most coagulation proteins is halted from mid-gestation until shortly before term onset of labor. The reason for this developmental arrest in plasma protein levels is unknown. The fetus demonstrates an unique balance of procoagulant, anticoagulant and fibrinolytic proteins concentrations which persists for several months postnatally. Fetal coagulation proteins are identical to adult proteins in structure and function with a few notable exceptions. Platelets are expressed early in fetal development and platelet concentrations are within the adult range by mid-gestation. Functional differences can be detected in fetal platelet function but rarely cause bleeding in the healthy fetus and newborn infant. Hemostasis in the fetus and newborn infants is functionally intact and spontaneous bleeding or clotting is rare. However, the neonatal hemostatic system appears to lack reserve capacity, and bleeding and clotting complications are common in the stressed infant, particularly the sick preterm infant. PMID- 15790158 TI - Arterial thrombosis in women. Pathogenesis. PMID- 15790159 TI - Postpartum haemorrhage. AB - Postpartum haemorrhage remains an important cause of maternal death in the developed and especially in the developing world. An appreciation of the physiological changes of pregnancy that predispose to rapid development of severe haemorrhage and DIC help maintain a level of vigilance. Although routine antenatal assessment can identify women with factors associated with an increased risk of severe postpartum haemorrhage, a significant proportion of women will develop intrapartum complications that cause severe haemorrhage. Prompt recognition and treatment of women with severe ongoing blood loss is essential to prevent morbidity and mortality. In addition to surgical correction of bleeding, replacement of plasma components to reverse coagulopathy and red cells to maintain tissues oxygenation are the basic aims of management. The haemostatic agent, recombinant Factor VIIa is a potentially useful addition to management of massive, life-threatening obstetric haemorrhage but its safety and efficacy remains untested in clinical trials. PMID- 15790160 TI - Hormone replacement therapy and arterial thrombotic disease. PMID- 15790161 TI - Thromboembolism in neonates and infants: impact of underlying diseases, prothrombotic risk factors and treatment modalities. PMID- 15790162 TI - Defective annexin A5 crystallization: a mechanism for pregnancy losses in the antiphospholipid syndrome. AB - We have accumulated a significant body of evidence that support the hypothesis that aPL antibody-mediated disruption of annexin A5 crystallization is a mechanism for RSPL in the APS. Further investigation of the functions of the annexins in placental physiology and disease are likely to be productive areas for future research. PMID- 15790163 TI - Mechanisms of OC related thrombosis. PMID- 15790164 TI - Monitoring of low molecular weight heparin (LMWH) in pregnancy. PMID- 15790165 TI - Assessment of placental flow. PMID- 15790166 TI - Hormone replacement therapy and venous thromboembolism. PMID- 15790167 TI - Thrombophilia and pregnancy failure: feto-maternal interactions in the vascular bed of the placenta. PMID- 15790168 TI - Prevention and treatment of arterial thrombosis in women. PMID- 15790170 TI - Thrombophilia and gestational venous thromboembolism. PMID- 15790169 TI - Maternal adaptation to vasculogenesis and angiogenesis at the feto-maternal interface. PMID- 15790171 TI - Pathogenesis of antiphospholipid syndrome. PMID- 15790172 TI - Organohalogen contaminants in delphinoid cetaceans. AB - This chapter reviews the global distribution, biotransformation, accumulation patterns, and mechanisms of action and the potential impacts of persistent organohalogen contaminants (PHCs) on physiological systems of cetaceans with emphasis on delphinoids. Methods used to study PHCs in stranded and free-living cetaceans are discussed, and concentrations of PHCs of stranded, hunted, by catch, and free-ranging delphinoids are summarized. Overall, the highest concentrations of PHC contamination were found in delphinoids from industrialized areas of the Northern Hemisphere compared to the Southern Hemisphere. Nonetheless, PHCs are also found in marine mammal tissues from the Southern Hemisphere and in remote regions such as the Arctic, reflecting the global distribution and contamination of PHCs in the marine ecosystem. PMID- 15790173 TI - Environmental contamination and human exposure to lead in Brazil. AB - Adverse effects caused by environmental lead pollution are well recognized. Being a widespread agent in the environment and a major harmful element to organic systems, mostly to children, lead has been investigated all over the world, aiming to improve measures regarding its control. The purpose of this chapter is to present a review of the situation of production, uses, assessment of exposure, and adverse effects from environmental lead contamination in Brazil. It also presents aspects of Brazilian legislation setting up maximum permissible levels of lead in several environmental compartments such as surface and drinking water, soils, sediment, urban air, and also in commercially sold food, vegetables, fish, and meat, in an effort to control industrial emissions. Epidemiological investigations on children's lead exposure around industrial and mining areas are revised, showing that many situations where lead contamination is potentially present still need to be addressed by governmental agencies. In Brazil, lead was withdrawn from gasoline by the end of the 1980s, and the last lead mining and primary smelting plant was closed in 1995, leaving residual environmental lead contamination, which has recently been investigated using a multidisciplinary approach. Nevertheless, there are hundreds of small secondary battery recycling plants all over the country, running smelting facilities that produce local urban areas of lead contamination. PMID- 15790174 TI - Arsenic speciation and toxicity in biological systems. AB - Although it is now commonly accepted that toxicity and bioavailability varies with As species, extensive research has been carried out on biological and environmental samples to assess toxicity and risk associated with As exposure based on total concentrations that may be in error. The health investigation guideline for the Australian environmental protection measure is 100 mg/kg (As(tot)), which would cause potential risk to human health if all the As present in a sample were bioavailable (ANZECC 1992). Similarly, the MPC for As in food is 1 mg/ kg (fresh weight), but this concentration may include contributions from As(III), AsV, and all organic species. Thus, a food substance, such as seafood, could have a high total concentration exceeding the guidelines, but most of the As would be in forms that are nontoxic to humans; i.e., the bioavailability is low, and the food would therefore be perfectly safe to eat. On the other hand, a food that has high bioavailability of As consequently is more toxic. Overall, it appears that contamination of water by As is probably more harmful to humans than As in food grains or vegetables, because As bioavailability in water is generally higher than its bioavailability in food. Nevertheless, As in food crops could make significant contribution toward total daily intake. Therefore, failure to consider the contribution of As species on their bioavailability could introduce a substantial bias into the estimation of risks associated with exposure as well as evaluation of As toxicity. In conclusion, As must be regarded as an important environmental toxicant because of its acute and chronic toxic properties and extensive presence in the environment. Much remains to be learned about its toxicology and biochemistry for better understanding of this important contaminant. PMID- 15790175 TI - Making the media work for us. PMID- 15790176 TI - Raising awareness of FGM. PMID- 15790177 TI - M is for midwifery. PMID- 15790178 TI - The pregnant woman's right to say no: a personal reflection. PMID- 15790179 TI - The heel prick: how efficient is common practice? AB - The heel prick is a common procedure, mainly carried out by midwives. This paper argues that midwifery practice is often based on tradition and habit rather than evidence. Firstly, the evidence on which midwives choose a site for heel prick is examined, and it is argued that practice is based on old evidence and that more recent research must be built on to ensure the best care for babies. It is argued that preparation of the heel is unnecessary, and that midwives should consider alternatives to squeezing the heel. Finally, it is argued that encouraging women to breastfeed their babies or otherwise maintain skin-to-skin contact throughout the procedure may be the most appropriate form of analgesia. PMID- 15790180 TI - Neonatal examination: are midwives clinically effective? AB - The objective of the pilot study was to test the appropriateness of an audit tool developed to determine the clinical effectiveness of midwives carrying out neonatal examination of the newborn. The audit was undertaken by reviewing retrospective data, collected from midwives' personal clinical records of neonates and the case records of the babies they had examined. The data was gathered over a period of 18 months, and included the records of 482 term babies. The setting was a district general hospital in the east of England. The participants were all midwives who had successfully completed the neurobehavioural examination of the newborn course, and practising at a local district general hospital. The findings of the study indicate that the audit tool was sufficiently robust to test the clinical effectiveness of midwives' detection of specific congenital abnormalities. Clinical effectiveness rates were reassuringly high. The study also began to highlight the positive impact on the improvement of the service available to women. A further study has been started to compare the clinical effectiveness rates between professional groups. PMID- 15790181 TI - The mutilated orchid. PMID- 15790182 TI - Child protection is everybody's business. PMID- 15790184 TI - Mastering a masters. AB - A need for greater theoretical knowledge and evidence on which to base their care led Annette Briley and Joy Kemp to undertake a Masters degree. They offer their personal experiences of balancing work, study and home life and encourage other midwives to apply for further education courses. PMID- 15790185 TI - The RCM staffing survey 2004. PMID- 15790186 TI - Leading the development of an ECV service. AB - This article has identified an area of practice that needed to be developed. Despite the service being a medical procedure, it fitted into the remit of my role. The findings provide a well-established with improving ECV clinical skills. The drive now will be to review the way women receive information about ECV (or ensure that they have the opportunity for a vaginal breech birth, if this is their choice). There is also the need to improve the chance of a vaginal birth following successful ECV. This initiative demonstrates a multidisciplinary team approach, working together to provide a service, which has been found to be safe and, above all, evaluated favourably by women. PMID- 15790187 TI - Should fathers be present during childbirth? PMID- 15790188 TI - The 'on-call' debate. PMID- 15790189 TI - [Survival of breast cancer patients treated with inhibitors of the aromatase vs tamoxifen]. AB - BACKGROUND: In Mexico the breast cancer occupies the second place as cause of death by oncological illness and the etiology is considered multifactorial. The systemic treatment of this cancer is necessarily after the surgery and simultaneously when radiotherapy is used; therefore the recent introduction of newly non toxics and efficient antiestrogens which block the effect of estrogens from circulation have become drugs of first line in the metastasis illness. OBJECTIVE: To compare the clinical evolution and the survival of patients with locally advanced breast cancer and metastasis with the use of two therapeutic modalities. MATERIAL AND METHODS: Under informed consent 20 patients with diagnosis of breast cancer and out of oncological therapeutic processing were included. Randomly an inhibitor of the aromatase (anastrozole, 1 mg for day) was used in 10 patients and in the other 10 patients tamoxifen (20 mg for day) both for 2 years, subject to verification of positive estrogens-progesterone receptors. RESULTS: Half of the patients of each group had a clinical stage III. The most frequent histological diagnosis was the ductal carcinoma in 70% of the group with tamoxifen use, while in anastrozole group was 90%; the mastectomy prior to the tamoxifen was carried out in 60% of the patients and in 50% of the group with anastrozole. The acceptable response to the processing in the patients with tamoxifen was complete in the 60% of the cases, but with the use of anastrozole was 80%; survival in the group with anastrozole was of 100% in two years of follow up, while in women that received tamoxifen the survival was 90% (p<0.001). CONCLUSIONS: The use of anastrozole improved the survival as well as the quality of life, showing no side effects in this group of patients. PMID- 15790190 TI - [Clinical experience with chemotherapy of the malignant tumor of germinal cells (dysgerminoma) of the ovary]. AB - OBJECTIVE: To evaluate two alternative plans of chemotherapy with EP (etoposide, platinum) and BEP (bleomycin, etoposide, platinum) after oncological surgery. MATERIAL AND METHODS: A clinical, longitudinal and descriptive study was done, where the cases with pure dysgerminoma diagnosed in the gynecology-oncology service during the years from 1992 to 2003 were included. Information was recollected in a precoding survey that included sociodemographic characteristics, tumor size, free survival of disease, general survival, recurrence index and the index of fertility. RESULTS: The age of the group studied was of 22.1 +/- 6.5 years, with stratification to 9 patients in phase I (50%), phase III in 7 patients (38.9%) and phase IV in 2 patients (11.1%). Histological study confirmed the pure dysgerminoma, with a mean duration of the symptoms of 5.2 months, the free survival of disease was of 39.6 months and general survival was of 49.5 months. It was administered early chemotherapy in 11 patients, from which 7 received EP and the other 4 received BEP. Eight patients were found in advanced phases and with metastasis. There were 4 (22.2%) recurrences, of which 3 corresponded to phase III and a case to phase IV with tumors all of them over 15 cm. There were 3 patients rescued with chemotherapy of second line and a patient with radiotherapy to central nervous system. The statistical analysis showed that size of tumor among the recurrent group (24.2 cm) and the group without recurrence (14.5 cm) had significant differences (p = 0.018), the size of the tumor and the free survival of disease did not have significant correlation (p = 0.99), but upon comparing the general survival with the phase of the disease, a significant correlation was found (p = 0.03), where the survival to 5 years was observed in 6 cases (33.3%), of which a case was treated with surgery and without chemotherapy, 4 cases with EP and a case with BEP. Out of the 11 cases treated with conservative surgery, two patients got pregnancy in three occasions. PMID- 15790191 TI - [Alcoholism during pregnancy: an underestimated health problem]. AB - OBJECTIVES: To identify the frequency of consumption of alcohol in pregnant women who went to a module of prenatal control; to describe the consumption habits and to identify the number of cases in those that the physician of first level identified the addiction, using the institutional instruments. MATERIAL AND METHODS: A descriptive and prospective study was carried out, the study population was selected by means of non randomized sampling of the total of pregnant women who went to receive services of prenatal control, in two units of family medicine of the Mexican Institute of Social Security, of Tapachula, Chiapas, Mexico. The instrument AUDIT (Alcoholism Disorders Identification Test) was used, to identify use-frequency, abuse, dependence and physical/mental damage conditioned by the alcohol. RESULTS: In 132 studied women, it found a frequency of 45.5% of pregnant women with positive consumption and a case of dependence, none of which was identified by the family doctor. CONCLUSIONS: The consumption of alcohol in the studied population is high, even bigger than the frequency detected in populations of non pregnant women. The use of detection tests such as the AUDIT and the implementation of measures guided to the training and the personnel's of health sensitization about the magnitude and impact of this problem are recommended. PMID- 15790192 TI - [Urolithiasis and pregnancy. Presentation of results and management norm at the National Institute of Perinatology]. AB - AIMS: Urolithiasis associated to pregnancy has an incidence of 1:1,500. It is more frequent in multiparity patients and is a risk factor for preterm labor in more than 40%. OBJECTIVES: To know the incidence of urolithiasis associated to pregnancy at the National Institute of Perinatology; to determine the prevalence of: symptoms and signs, hydronephrosis, urinary tract infections, uropathogens, use of catheters and perinatals results. METHODS: A cohort study was made from January 1998 to October 2003, to identify pregnant patients with urolithiasis. The clinical files were reviewed; Fisher's exact test was used for statistical analysis. RESULTS: Twenty-one patients were diagnosed with urolithiasis; 71% had lumbar pain; 57.1% hematuria; 47.6% bilateral urolithiasis; and 33% bilateral hydronephrosis. All the patients who had lumbar pain, hematuria or were catheterized developed urinary tract infection, even with antibiotics prophylaxis. The 52.4% had at least one urinary tract infection episode. The average gestational age at birth was 38.5 weeks (+/- 5.6 weeks). The 84.22% were preterm births. CONCLUSION: Our incidence is similar to the one reported in the literature. The 71.26% has at least one of the symptoms from the classic triad of urolithiasis. The prevalence of urinary tract infection in these patients is very high therefore; antibiotics prophylaxis and urine cultures are required. A lower prevalence of preterm births was obtained. A flow chart for diagnosis and follow up was presented in order to decrease and to face the possible complications of this pathologic entity. PMID- 15790193 TI - [Predisposing factors to breast cancer in the Region Langunera (Mexico)]. AB - OBJECTIVE: To identify the presence and frequency of various predispose factors of breast cancer in the Region Lagunera (Mexico). PATIENTS AND METHODS: 38 women with breast cancer (Group A, cases) and 24 women without breast pathology (Group B, controls) were analyzed; throughout a questionnaire the variables that have been described as predisposition factors were researched. Group A included patients from the Instituto Mexicano del Seguro Social, Speciality Hospital # 71, Oncology Department in Torreon, Coahuila, from May 1st to October 31st of 1999. At the same time 24 women (Group B), who attended to have an ultrasound and/or a mammography practice in a private clinic in the same city, were polled having a normal result. The means of the two treatments were compared using the SAS program version 6.3. RESULTS: The following variables showed significant difference: mother and father family background with cancer p=0.032; previous breast pathology p=0.0032; hormone consumption p=0.0289; lactation p=0.038 and time of lactation p=0.036. CONCLUSION: Overweight, obesity, mother and father family background with cancer, previous breast pathology background, period of consuming hormone, lactation and lactation period had significance in this study. PMID- 15790194 TI - [Inauguration speech as a president of the Mexican Association of Gynecology and Obstetrics (2004-2005)]. PMID- 15790195 TI - [Symphysiotomy. 1949]. PMID- 15790196 TI - Factors promoting rupture of abdominal aortic aneurysms. AB - The risk for rupture of an abdominal aortic aneurysm is widely believed to be related to its maximum diameter. Rupture occurs at the site of maximum wall stress, when it exceeds the tensile strength of the aortic wall. Basic research confirmed that peak wall stress and aortic wall biodegradation contribute to the mechanism of aneurysm rupture. In order to highlight the role of loss in wall strength and increase in focal peak stress, the authors reviewed recent literature. The clinical relevance of these recent insights in the etiopathogenesis of aneurysm rupture is analysed. PMID- 15790197 TI - Endovenous therapy--2005. PMID- 15790198 TI - Diagnosis of deep vein thrombosis: how many tests do we need? AB - The requirement for a safe diagnostic strategy should be based on an overall post test incidence of venous thromboembolism (VTE) of less than 1% during 3 month follow-up. Compression ultrasonography (CUS) has a negative predictive value (NPV) of 97 to 98% indicating a post-CUS incidence of deep vein thrombosis (DVT) of 2 to 3%. A post-CUS DVT incidence of 3% implicates that 90 to 120 DVTs per 1 million inhabitants will be overlooked each year indicating the need to improve the diagnostic work-up of DVT as much as possible. The qualitative D-dimer test (SimpliRed) has a sensitivity of 82 to 89% and a negative predictive value of 94 to 95% indicating a 5 to 6% post-test incidence of DVT, which is not sensitive enough for venous thrombosis exclusion. The post-test DVT incidence could be reduced from 3.2% to 0.6% in one study and from 11% to 2% in another study by the combination of a normal CUS and low clinical score and from 4.5% to 1.6% by the combination of low clinical score and a negative SimpliRed test in one study. The combination of a negative CUS and a negative SimpliRed test reduced the post-test incidence of DVT from 2.6% to < 1% or even < 1% in two management studies without the need of a repeated CUS on the basis of which anticoagulant therapy can safely be withheld. The rapid quantitative turbidimetric D-dimer assay (Tinaquant) has a sensitivity and a negative predictive value (NPV) of 97.7% with a 2.3% post-test incidence of DVT. The combination of a normal Tinaquant D-Dimer test result plus a low to moderate clinical score reduces the post-test incidence of DVT from 2.3 to 0.6% without the need of CUS testing in 29% of patients with suspected DVT. The rapid ELISA VIDAS D-dimer assay has a sensitivity and NPV of 98.6 and 99.5% in two management studies for the exclusion of DVT irrespective of clinical score. The combination of a normal ELISA VIDAS D-Dimer test with clinical score assessment will reduce the post-test DVT incidence of less than 0.5% and the need for CUS testing by 40 to 50%. It is concluded that the sequential use of a rapid quantitative D-dimer test, clinical score and CUS appears to be safe and the most cost-effective diagnostic work-up of DVT. PMID- 15790199 TI - Non-invasive diagnosis of pulmonary embolism, anno 2005. AB - Pulmonary angiography is the gold standard for segmental pulmonary embolism (PE) but no longer for subsegmental PE, because the inter-observer agreement for angiographically documented subsegmental PE is only 60%. Two non-invasive tools exclude PE with a negative predictive value of >99% : a normal perfusion lung scan and a normal rapid ELISA VIDAS D-dimer test. The positive predictive value is 85 to 88% for a high probability ventilation-perfusion lung scan (VP-scan) and >95% for helical spiral CT. The prevalence of PE in management studies of symptomatic patients with a non-diagnostic VP-scan is 20 to 24%. Helical spiral CT detects all clinically relevant PE and a large number of alternative diagnoses in symptomatic patients with a non-diagnostic VP-scan or a high probability VP scan. Single-slice helical CT as the primary diagnostic test in patients with suspected PE in three retrospective studies and in two prospective management study indicate that the negative predictive value of a normal helical spiral CT, a negative compression ultrasonography of the legs (CUS) together with a low or intermediate pre-test clinical probability is >99%. Therefore, helical spiral CT can replace both the VP-scan and pulmonary angiography to safely rule in and out PE. The combination of clinical assessment, a rapid ELISA VIDAS D-dimer followed by CUS will reduce the need for helical spiral CT by 40 to 50%. PMID- 15790200 TI - New ways to prevent venous thromboembolism--the pentasaccharide fondaparinux and the thrombin inhibitor ximelagatran--a review. AB - During the last decade low molecular weight heparins have been the dominating methodology to prevent postoperative venous thromboembolism. They are effective and safe but in high risk surgery (major orthopaedic and abdominal/pelvic cancer) there is still a significant thrombotic problem. Recently two new thromboprophylactic principles have appeared--one Xa inhibitor in the form of the pentasaccharide fondaparinux and one direct thrombin inhibitor in the form of melagatran with an orally absorbable prodrug--ximelagatran. Both have been evaluated in extensive research programmes and both have been approved for use in major orthopaedic surgery by the European health care authorities. PMID- 15790201 TI - Psychosocial effects of bariatric surgery. AB - INTRODUCTION: Morbid obesity is associated with an increased risk of morbidity and mortality as well as invalidating psychosocial problems and poor quality of life. The eventual goal of bariatric surgery is not only reduced weight, but also improved quality of life. What are these psychosocial consequenses of bariatric surgery? MATERIAL AND METHODS: A systematic literature search identifying relevant psychosocial consequences of bariatric surgery. RESULTS: Bariatric surgery does not only lead to substantial weight reduction, but also to improvement or cure of comorbidities, including diminishing of psychopathology and abnormal eating behavior, and improvement in quality of life. However, a significant minority of morbidly obese individuals do not respond positively to surgery and do not benefit psychologically from surgery, also mixed results on long-term quality of life are reported. DISCUSSION: The psychosocial effects of bariatric surgery, being significant improvement in psychosocial functioning and quality of life, provide additional justification for the surgical treatment of morbid obesity. PMID- 15790202 TI - Long-term results after excision haemorrhoidectomy versus stapled haemorrhoidopexy for prolapsing haemorrhoids; a Belgian prospective randomized trial. AB - PURPOSE: To compare the postoperative evolution and the long-term efficacy after stapled haemorrhoidopexy (PPH) and Milligan-Morgan haemorrhoidectomy (MM). METHODS: In a prospective randomized study, 40 patients requiring surgical treatment for prolapsing haemorrhoids grade II or III were assigned to either MM or PPH (20 each). Postoperative pain, wound healing were evaluated, as well as anal pressures and sphincter anatomy. Mean follow-up is 46 months. RESULTS: Postoperative pain at rest and during defecation was less important after PPH if no resection of external piles or skin tags was associated (P < 0.0001). Healing time was shorter after PPH (P < 0.0001). Endoanal ultrasound remained unchanged postoperatively. Resting and squeeze pressures decreased after MM, but not after PPH (P < 0.01). After a mean follow-up of 46 months (12-56), persistent or recurrent symptoms, mostly mild and temporary, were observed after both MM and PPH, in 7 and 11 patients respectively (NS). After PPH, five patients (25%) complained of recurrent external swelling and/or prolapse (P = 0.047 vs. MM) requiring redo surgery in four of them, after 10, 13, 14 and 21 months. No redo surgery was required after MM. Long term patient satisfaction after PPH was not better than after MM. CONCLUSIONS: Postoperative pain is less important after PPH. This advantage disappears if any resection is associated with the stapling. At medium to long-term follow-up, PPH seems to carry a higher risk of symptomatic external haemorrhoidal disease, needing further surgery. PMID- 15790204 TI - Metastasis to the breast--a clinical challenge in outpatient. AB - Metastatic tumours of the breast are often misdiagnosed as a primary breast malignancy. This erroneous diagnosis can lead to unnecessary mutilating surgery and to delays in applying appropriate treatment. We have analyzed the clinical features of seven patients presenting initially with a breast mass which, after further investigation, proved to be metastasis from a non-breast primary tumour. The treatment of metastatic breast tumours depends on the nature of the primary tumour. Metastases of solid tumours are treated with local excision. Lymphomas are treated with appropriate systemic therapy. The prognosis for patients with metastatic tumours in the breast is poor. PMID- 15790203 TI - Acute rehabilitation program after laparoscopic colectomy using intravenous lidocaine. AB - BACKGROUND: The concept of postoperative acute rehabilitation was introduced to accelerate postoperative recovery and improve outcome. We investigated whether intravenous lidocaine infusion, which decreases postoperative pain and speeds the return of bowel function, can be used instead of epidural analgesia in an acute rehabilitation protocol for patients undergoing laparoscopic colectomy. METHODS: Twenty eight consecutive patients scheduled for laparoscopic colectomy were prospectively included in this case series study. Segmental colectomy was performed only for benign pathology. Intraoperative opioid use was restricted. After a bolus injection of lidocaine 1.5 mg x kg(-1), an infusion (2 mg x kg(-1) x h(-1), i.v.) was started before pneumoperitoneum. Balanced analgesia was used to reduce postoperative opioid consumption. Patients were allowed to drink 6 h postoperatively. The day after surgery, patients were allowed to eat a normal breakfast. Enforced mobilisation and ambulation were required from the patients. Our goal was to discharge patients within 3 days after surgery. Postoperative pain was measured. Time to first flatus, defecation, and hospital discharge were recorded. RESULTS: Mean postoperative pain at rest and mobilisation remained below 30 mm on a 100 mm visual analogue scale. Time to first flatus, defecation, and hospital discharge were 29 +/- 13 h, 38 +/- 13 h, and 3.0 +/- 1.0 days, respectively. CONCLUSION: Acute rehabilitation after laparoscopic colectomy using i.v. lidocaine gives similar outcomes to those reported using epidural analgesia. PMID- 15790205 TI - Breast conserving surgery after primary chemotherapy in locally advanced breast cancer. AB - PURPOSE: Primary chemotherapy is being given in the treatment of locally advanced breast cancers (LABC), but a major concern is local recurrence after therapy. The aim of this study was to assess the role of breast conserving surgery (BCS) in patients with locally advanced breast cancer. MATERIAL AND METHODS: Twenty-eight patients, presenting LABC (T any, N 012, M0) were treated with primary chemotherapy comprising of cyclophosphamide, doxorubicin and fluorouracil and then BCS followed by radiotherapy were examined between the years 1992-2002 retrospectively. Before neoadjuvant chemotherapy, seven patients (25%) were Stage IIB, 19 patients (68%) Stage IIIA and two patients (7%) Stage IIIB. Survival times and curves were established according to the Kaplan-Meier method and compared by means of the log-rank test. The chi-square test and log rank test were performed for univariate statistical analysis of each prognostic factor. P values in multivariate analysis were carried out by the Cox's proportional hazards regression model. All p values were two-sided in tests and p values <0.05 were considered significant. RESULTS: Clinical down staging was obtained in 25 (89%) of patients. Three (11%) patients had complete clinical response, 22 (78%) patients with partial response and 3 (11%) had stable disease. The primary tumour could not be palpated after chemotherapy in 6 (21%) of 28 patients presenting with palpable mass, therefore needle localization was performed for BCS. Median follow-up was 51.9 months (ranging 10 to 118 months). Local recurrence was detected in 4 (14%) patients. Distant metastasis developed in 5 (18%) patients. Three of the patients died of distant metastases and two of them are alive at 49 months. Five-year survival rate was 66%. Statistically, there were no significant factors in terms of local recurrence. Histological grade and menopause status were significantly associated with overall survival (p = 0.018) and nuclear grade was the one significant factor on distant disease-free survival in univariate analysis (p = 0.006). In multivariate analysis, there were no significant factors in terms of overall and distant disease-free survival CONCLUSIONS: Negative margin is more important than the clinical and histological parameters, such as pretreatment stage, clinical response rate, ER and PR in terms of local recurrence. BCS can be performed safely by achieving free surgical margin in patients who have small sized tumour and with either N2 axillary involvement or skin invasion. PMID- 15790206 TI - Experience with laparoscopic adjustable gastric banding (la-band system) up to 8 years. AB - Morbid obesity occurs in 2-5% of the population in Western countries. Laparoscopic adjustable silicone gastric banding is a minimally invasive, adjustable and reversible procedure for the treatment of morbid obesity. The lap band system was evaluated retrospectively in a series of 222 patients. Postoperative outcome and weight loss patterns at up to 8 years follow-up are presented. The most frequent late complications were a leak between the port and the catheter, which occurred in 21 patients (9.4%) and total and irreversible food intolerance due to pouch dilation and/or slippage, which occurred in 13 patients (5.8%). The postoperative BMI reductions are successful and stable after a follow-up of up to 96 months. The lap-band system seems an effective procedure for achieving appreciable and stable weight loss up to 8 years of follow-up and the complications and re-operation rates are acceptable. In 81% of the cases also, the patient is very satisfied with the results of the operation. From the 47.3% who found their quality of life before the operation bad or even devastating, 93% envoy life after the operation like never before. PMID- 15790207 TI - Strictures of the terminal bile duct--who profits from the surgery? AB - BACKGROUND: Elucidation of the etiology of terminal bile duct strictures is oftentimes challenging. In choosing the optimal treatment method, a multidisciplinary approach is necessary. AIMS AND METHODS: To demonstrate the advantages and indications of surgical management, 153 patients with terminal bile duct strictures were retrospectively analysed. All patients had been treated during a period of eight years (1st January 1995 - 31st December 2002) in same department. The principal datas studied were surgical morbidity, thirty-days' mortality rates and recurrence of jaundice. RESULTS: During a period of eight years, 153 patients (100%) underwent laparotomy. Radical surgery (duodenopancreatectomy) was performed in 79 patients (51.6%), while 74 others (48.4%) benefited from bypass procedures. Postoperative morbidity reached 13.1%; nine patients (5.9%) died within the thirty days of surgery. Recurrence of jaundice was observed in three patients (1.9%). CONCLUSION: Surgical management of terminal bile duct strictures is indicated in most patients with benign strictures, strictures of unknown origin and with malignant strictures, where the patients are in a reasonably satisfactory clinical condition. The surgical approach provides the potential advantage to obtain a definite histological diagnosiscare and to remove the tumour in many patients. Patients with non resectable tumours could benefit from bypass procedures ensuring long-term and effective drainage of the bile. PMID- 15790208 TI - Chronic lunotriquetral ligament injuries: arthrodesis or capsulodesis. AB - Lunotriquetral ligament ruptures were treated by arthrodesis or ligament reconstruction. The outcomes were retrospectively reviewed: 17 underwent an arthrodesis, 13 a reconstruction with a strip of the extensor retinaculum. In the arthrodesis group, nine were not satisfied; complications and re-operations were numerous; eight developed a pseudarthrosis. In the reconstruction group, eight were satisfied, five were not. Complications were less frequent and less severe. PMID- 15790209 TI - Traumatic lesions of the diaphragm. Our experience in 33 cases and review of the literature. AB - We reviewed 33 consecutive patients with diaphragmatic injuries. Twenty-nine were admitted in emergency conditions after blunt (22 patients) or penetrating injury, presenting shock, dyspnoea, coma or acute abdomen in 21 cases; major associated lesions were found in 23 patients. Four patients presented acute complications of visceral herniation 2, 4, 84 and 216 months after the trauma. The diagnosis was preoperative in 23 cases, intraoperative in 9; in one case it was missed at laparotomy, becoming evident the day after. The sensibility of preoperative chest x-ray and CT was 86% and 100% in presence of visceral herniation, 14% and 0% in absence of visceral hernia. The diaphragmatic repair was always obtained by direct suture, following 20 haemostatic procedures (liver, spleen, mesenterium) and two bowel resections. The mortality rate was 24.4%; the morbidity rate was 48%. Traumatic lesions of the diaphragm are generally expression of particularly severe trauma whose outcome is mainly influenced by the associated lesions. They are also correlated to specific morbidity and mortality, so the surgical exploration is mandatory whenever this injury is suspected, considering that the preoperative diagnosis relies on visceral dislocation. Associated lesions influence the surgical strategy but a direct suture is usually effective in preventing specific complications. PMID- 15790210 TI - Is laparotomy necessary in the diagnosis of fever of unknown origin? AB - Fever of unknown origin (FUO) is a diagnostic challenge for the practising physician. Detailed medical history, physical examination, non-invasive laboratory tests, and radiologic examinations compose the first level in the diagnostic approach to the FUO. When a diagnosis cannot be established with these procedures, some invasive diagnostic techniques and finally exploratory laparotomy are performed. Although advanced diagnostic measures and imaging guided less invasive procedures have decreased the need, laparotomy remains as a final diagnostic method for FUO cases. In this study we evaluate the role and importance of laparotomy in the diagnosis of our FUO cases. In 17 out of 126 patients (8 male, 9 female, the median age 35.8 years) hospitalized in our clinic between 1982 and 2002 with the diagnosis of FUO, the diagnosis was established by laparotomy. The diagnosis was made directly in 13 patients, and indirectly (by excluding other diseases) in 2 patients. In several FUO series, the contribution of laparotomy to the diagnosis of FUO was reported as 27-100%. This rate was found to be 88% in the present study. During laparotomy on 17 cases, tissue samples were taken from spleen, liver, intra-abdominal and mesenteric lymph nodes. Pathologic examination of these tissue samples revealed miliary tuberculosis in 4; non-Hodgkin's lymphoma in 3; Hodgkin's lymphoma in 3; liver tumour in 1; hairy cell leukemia in 1; peritonitis carcinomatosis in 1. In the patients with miliary tuberculosis, the liver (3) and/or spleen (2), and/or lymph node (3) revealed caseating granulomas. Laparotomy diagnosed 3 of 5 cases whose abdominal ultrasonography and computerized tomography were normal. In conclusion, although advanced diagnostic methods decreased the need for laparotomy in FUO, if non-invasive and invasive diagnostic measures fail, laparotomy may contribute to the diagnosis. The selection of the patient and the timing are important for laparotomy. PMID- 15790211 TI - Abdominal aortic aneurysm due to Brucella melitensis. PMID- 15790212 TI - Life saving abdominal decompression in a patient with severe acute pancreatitis. AB - Severe acute pancreatitis can be complicated early in its course by life threatening conditions such as abdominal compartment syndrome. We report a patient who needed abdominal decompression three days after admission to the intensive care unit because of intra-abdominal hypertension and end stage organ dysfunction. The clinical course was protracted, but the patient survived and was discharged from the hospital. PMID- 15790213 TI - Cancer of ileostomy: a late complication of colectomy for ulcerative colitis. AB - Cancer arising at an ileostomy site represents a rare late complication of total colectomy performed for ulcerative colitis. There are no more than 36 published cases in the literature. We describe a case of adenocarcinoma of the mucocutaneous junction at the ileostomy site, occurring 31 years after total colectomy for ulcerative colitis. Wide excision of the moderately differentiated adenocarcinoma was performed with refashioning of the ileostomy. Polypoid adenomas or adenocarcinomas of the ileostomy after colectomy performed for non neoplastic conditions are extremely uncommon. Biopsies of polypoid lesions at the stoma site are recommended. PMID- 15790214 TI - Metastatic thymoma as a retrovesical mass. AB - Retrovesical masses can be of various histopathological origin, most often prostatic. Primary tumours as well as metastatic lesions should be considered in the differential diagnosis. Metastatic thymoma, as presented in this case report, can be an extremely rare cause of retrovesical space occupying lesion. PMID- 15790215 TI - Acute respiratory failure from tracheopathia osteoplastica. AB - Tracheopathia osteoplastica is a benign cartilaginous and osseous metaplasia of the laryngo-tracheobronchial tree diagnosed more commonly in adults over 50 years of age. We report here the case of a 54-year-old man who underwent thyroidectomy for multinodular goiter. Immediately after an uneventful surgery, he developed an acute respiratory failure with radiologic picture of Adult Respiratory Distress Syndrome. Mechanical ventilation was set up again, bronchoscopy with biopsy disclosed a massive tracheobronchial haemorrhage from a tracheopathia osteoplastica. Supportive treatment was successfully provided and the patient resumed to a normal life 14 days after the operation. PMID- 15790216 TI - Massive ovarian oedema in adolescents. AB - Massive ovarian oedema is an unusual clinical entity occurring in teenage girls. It is a rare tumour-like condition of the ovary characterized by marked enlargement of one or both ovaries due to marked accumulation of oedema fluid in the ovarian stroma. Massive ovarian oedema may result from partial or intermittent torsion of the mesovary, interfering with venous and lymphatic drainage, but not with arterial blood flow. The patients most commonly present with lower abdominal pain and enlargement. This entity can clinically mimic a neoplasia. We report three cases with massive ovarian oedema. PMID- 15790217 TI - Colloid carcinoma arising in an anorectal fistula in Crohn's disease: a case report. AB - Carcinoma arising in a chronic anorectal fistula in Crohn's disease is rare, but the association has been reported in the literature. In most cases, it concerns a colloid carcinoma. Probably chronic irritation at either end of a fistula can trigger the degeneration of scar tissue into cancer. The diagnosis is difficult, due to lack of specificity of symptoms and signs, and is often delayed, resulting in a poor prognosis. A 70-year-old female, diagnosed with Crohn's disease at the age of 45, developed a pararectal colloid carcinoma in an anorectal fistula that had existed for years. She was treated with an abdominoperineal resection and adjuvant chemotherapy (5-fluorouracil and levofolinate) but died. A high index of suspicion and regular surveillance is recommended in chronic anorectal fistulas in Crohn's disease. In case of doubt, repeated biopsies should be performed to rule out malignancy. PMID- 15790218 TI - Granular cell tumour of the perianal region: which therapeutic attitude? AB - Granular cell tumours are uncommon soft tissue tumours that could be located anywhere throughout the body. Most of time they are benign but the diagnosis of malignancy is difficult to establish on histology: this is why the therapeutic decision is not easy to take in particular for some locations. We report the case of a 18-year old man: he presented, in the perianal region, a granular cell tumour of uncertain malignant potential (so far the first case described in the literature) for which we decided a conservative surgical treatment that failed. PMID- 15790219 TI - Laparoscopic cholecystectomy in a patient with situs inversus using ultrasonically activated coagulating scissors. Report of a case and review of the literature. AB - Situs inversus totalis is a rare defect with a genetic predisposition, which can present difficulties in the management of abdominal pathology, especially in laparoscopic surgery, due to the mirror-image anatomy. Herein, we report on a patient with situs inversus totalis and symptomatic cholelithiasis. The patient had a successful laparoscopic cholecystectomy, using ultrasonically activated coagulating scissors. Diagnostic pitfalls and technical details of the laparoscopic cholecystectomy are discussed and a review of the relevant literature is presented. PMID- 15790220 TI - Chronic adhesive small bowel volvulus: "whirl sign" on CT-scan. PMID- 15790221 TI - Editorial: Novatech 2004--synthesis of 3 years of scientific and technical advances in urban water management. PMID- 15790222 TI - Statistical analysis of pollution in stormwater infiltration basins. AB - In order to characterize soil pollution in stormwater infiltration basins, four basins of the Lyon area were selected and sampled. Altogether, 66 soil samples were collected. Each sample was analyzed for pH, cationic exchange capacity, particle size distribution, total organic carbon, total nitrogen, total phosphorus, metals, total hydrocarbons and polycyclic aromatic hydrocarbons (PAHs). Data was explored in a classic way (soil profiles) and using multivariate analysis techniques. Two methods were applied: Principal Component Analysis (PCA) and Cluster Analysis (CA). Main results are presented in this paper. Pollutant concentrations decrease rapidly with depth while pH and grain size increase. Sustainable concentrations are reached at a 50-cm depth, even after 21 years of operation. Multivariate analysis shows how pollution affects each sampling depth. PMID- 15790223 TI - Metal concentrations in soil and seepage water due to infiltration of roof runoff by long term numerical modelling. AB - The qualitative effects of stormwater infiltration on soil and seepage water are investigated with long term numerical modelling. The retention behaviour of different soils and materials used in infiltration devices is determined with batch and column tests. Results of the laboratory tests are adsorption isotherms which represent input data for numerical transport modelling. The long term simulations are performed with combinations of different solutions (types of roof runoff) and infiltration devices (swale and trench) under different hydrogeological conditions. The presented results contain the infiltration of low polluted roof runoff, runoff from a roof with zinc sheets and from a roof with copper sheets concerning the heavy metals zinc, copper and lead. The increase of concentrations in the infiltration body is high. For the infiltrated water, the results show a migration to groundwater only for the low adsorbing soil. PMID- 15790224 TI - Assessment of biofilm destabilisation and consequent facilitated zinc transport. AB - In infiltration basins, such as in any kind of porous media, bacteria may form biofilms. When conditions induce destabilization of this biofilm, resulting colloids are transported by the hydraulic flow. Some studies have focused on the role played by these bacterial colloids in pollutants transport in soil. This study deals with the ability of Pseudomonas putida to retain zinc and investigates the facilitated transport of this metal. Batch and columns experiments have been carried out. Bacteria display a great affinity for zinc in batch experiments and facilitated transport have been highlighted in a small extent, for the moment. A scenario of stabilization/destabilization of the biofilm has been designed and may be employed for further investigations. PMID- 15790225 TI - Can geotextiles modify the transfer of heavy metals transported by stormwater in infiltration basins? AB - Geotextiles are fibrous materials increasingly employed for the design of infiltration basins. However, their influence on the transfer of contaminants carried by stormwater has not been fully investigated. This study, based on column leaching experiments, aims at showing the effect of geotextiles on the transfer of three heavy metals (Zn, Pb and Cd) in a reactive soil (simulating an infiltration basin at laboratory scale). This effect depends on several factors, such as type of geotextile, hydric conditions (geotextile water content), hydraulic conditions (flow-rates) and the number of geotextiles installed. In all cases, geotextiles influence heavy metal retention by modifying flow and thus regulating contact between these metals and the reactive soil. PMID- 15790226 TI - Effective pollutant retention capacity of permeable pavements for infiltrated road runoffs determined by laboratory tests. AB - The infiltration of urban runoff always implies an entrance of pollutants into the soil and ground water. Due to legal regulations in many communes there is no longer any permission needed for stormwater infiltration, if administrative regulations and the requirements of standards are observed. The results of a research project carried out under the heading "Development of an assessment procedure for permeable pavements" show, that the pollutant retention capacity of permeable pavements varies considerably, depending on the material and the specific reactive surface. The objective of the study was to work out recommendations of suitable permeable pavements for different types of urban runoff. Selected data about the quality of urban runoff was compiled into a runoff matrix, which was used for defining characteristic dilutions. In batch tests, the material of the infiltration devices is penetrated with the dilutions. A test installation in large scale is used to calibrate the sorption coefficients derived from the batch experiment. PMID- 15790227 TI - Chemical hazard identification and assessment tool for evaluation of stormwater priority pollutants. AB - Assessment of chemical hazards is a critical issue, which have to be dealt with when evaluating different strategies for sustainable handling of stormwater. In the present study, a methodology for identifying the most critical and representative chemical pollutants was developed. A list of selected stormwater priority pollutants (SSPP-list) is the out-put from the procedure. Two different strategies for handling of stormwater were considered; discharge into a surface water recipient and infiltration. However, the same methodology can be used for other types of wastewater and other strategies for handling and treatment. A literature survey revealed that at least 656 xenobiotic organic compounds (XOCs) could be present in stormwater. In the next step, 233 XOCs were evaluated with respect to the potential for being hazardous towards either aquatic living organisms or humans, or causing technical or aesthetical problems. 121 XOCs were found have at least one of these negative effects, while 26 XOCs could not be assessed due to the lack of data. The hazard assessment showed that 40 XOCs had a PEC/PNEC ratio above one., e.g. they should be considered as priority pollutants. The final step is the expert judgement, which resulted in a final SSPP-list containing 16 selected priority pollutants. PMID- 15790228 TI - Wastewater planning in Djenne, Mali. A pilot project for the local infiltration of domestic wastewater. AB - The city of Djenne (Mali) is famous for its mosque which is the world's largest adobe building. Djenne is considered to be one of the most interesting cities in Western Africa from an architectural point of view. Since 1988, Djenne is listed as a UNESCO World Heritage Site. In the 1990s, a drinking water supply network was provided by foreign development organisations. However, no facilities were provided for the disposal of wastewater, resulting in an increase of waterborne diseases. This paper describes the urban drainage problem in Djenne and discusses the systematic evaluation of possible solutions and the subsequent pilot project for the local infiltration of sullage. The infiltration of sullage proved to be an appropriate technology for solving the urban drainage problem in Djenne. PMID- 15790229 TI - Experimental abatement data of underflow baffles for removal of floatables in the CSOs of the Greater Montreal (Canada) area. AB - Underflow baffles have gained in popularity over the years as a viable mean to intercept floatables in Combined Sewer Overflows (CSOs). This choice was mainly justified by the extremely low capital cost (CAPEX) and operational cost (OPEX) of this solution, although the efficiency of underflow baffles has never been clearly proven. The only similar application to underflow baffles are scum boards in grit chambers and clarifier. However, the flow conditions at CSOs vary considerably from those in grit chambers and clarifier. For this reason, review of the behavior of floatables in a rapid flow is paramount. Only then can comprehensive design criteria for underflow baffles and overflow chambers be suggested. Pilot scale tests, performed in a 17 metres long basin at various flowrates, had already shown that a critical horizontal velocity for floatables (Vcr) may develop in the overflow chambers. In this follow up study, the fate of intercepted floatables was investigated. It appears from this latest data that permanent capture of floatables decreases rapidly with an increase in the horizontal velocity of the flow, no matter what the baffle depth. Baffle depth increases capture at lower velocities (0.17 m/s) but become irrelevant at higher velocities (0.61 m/s). This data suggests that capture efficiency of existing underflow baffles in overflow chambers can be, at best, very low whenever the horizontal velocity increases above 0.30 m/s or 1 ft/s. PMID- 15790230 TI - Solids separation efficiency of combined sewer overflows. AB - The removal of sewer solids at combined sewer overflow locations depends on the flow patterns inside the overflow structure on the one hand and on the sediment characteristics on the other hand. Flow conditions can be described by the residence time distribution; sewer sediments can be characterised by their settling velocity distribution. The combination of both distributions leads to a dimensionless efficiency curve, which gives the removal efficiency as a function of the Hazen number. For field conditions this efficiency curve is mainly influenced by the settling velocity distribution of the sewer sediments and, as a consequence, nearly identical efficiency curves are found for different types of prototype CSO structure. For design purposes, a methodology using return frequency analysis is proposed. PMID- 15790232 TI - The influence of biodegradability of sewer solids for the management of CSOs. AB - The re-suspension of sediments in combined sewers and the associated pollutants into the bulk water during wet weather flows can cause pollutants to be carried further downstream to receiving waters or discharged via Combined Sewer Overflows (CSO). A typical pollutograph shows the trend of released bulk pollutants with time but does not consider information on the biodegradability of these pollutants. A new prediction methodology based on Oxygen Utilisation Rate (respirometric method) and Erosionmeter (laboratory device replicating in-sewer erosion) experiments is proposed which is able to predict the trends in biodegradability during in-sewer sediment erosion in wet weather conditions. The proposed new prediction methodology is also based on COD fractionation techniques. PMID- 15790231 TI - High-rate stormwater clarification with polymeric flocculant addition. AB - Treatment of urban stormwater by clarification, with flocculant addition, was studied in Toronto, Canada using a pilot-scale clarifier with removable lamellar plates. Almost 90 stormwater runoff events were characterised at the study site and found fairly polluted. The previous research phase indicated good treatability of this stormwater by lamellar clarification with flocculant addition (total suspended solids, TSS, removal of 84%, at a surface load of 15 m/h), but there were concerns about cleaning plates after storm events. With the aid of numerical modelling, hydraulic improvements to the clarifier inlet zone were retrofitted in 2004 and permitted the removal of the lamellar pack without a loss in treatment efficiency. In the modified clarifier, a cationic polymeric flocculant dosage of 4 mg/L with conventional clarification provided a TSS removal of 77%, at surface loads up to 43 m/h. The use of the polymer did not increase the acute toxicity of the treated effluent. The clarifier sludge was severely polluted by several heavy metals and would require special disposal. The treatment process tested could be well applied in projects requiring intensive stormwater treatment at compact sites. PMID- 15790233 TI - Performance-costs evaluation for urban storm drainage. AB - The design process of urban stormwater systems incorporating BMPs involves more complexity unlike the design of classic drainage systems for which just the technique of pipes is likely to be used. This paper presents a simple decision aid methodology and an associated software (AvDren) concerning urban stormwater systems, devoted to the evaluation and the comparison of drainage scenarios using BMPs according to different technical, sanitary, social environmental and economical aspects. This kind of tool is particularly interesting so as to help the decision makers to select the appropriate alternative and to plan the investments especially for developing countries, with important sanitary problems and severe budget restrictions. PMID- 15790234 TI - Performance indicators for the efficiency analysis of urban drainage systems. AB - Performance indicators implemented in a decision support system (DSS) for the technical, managerial and economic evaluation of urban drainage systems (UDS), called MOMA FD, are presented. Several kinds of information are collected and processed by MOMA FD to evaluate both present situation and future scenarios of development and enhancement. Particular interest is focused on the evaluation of the environmental impact, which is considered a very relevant factor in the decision making process to identify the priorities for UDS improvements. PMID- 15790235 TI - A new database on urban runoff pollution: comparison of separate and combined sewer systems. AB - For a long time people have questioned what the "best" sewer system is for limiting the pollution load released into the receiving waters. In this paper the traditional separate and combined sewer systems are compared using a pollution load balance. The investigation is based on measured concentration data for a range of pollutant parameters in the sewer from the new database "ATV-DVWK Datenpool 2001". The approach also accounted for the wastewater treatment plant outflow which contributes to the total pollutant load considerably. In spite of a number of neglected effects, the results show that the separate system is superior to the combined for some parameters only, such as nutrients, whereas for other parameters, e.g. heavy metals and COD, the combined system yields less total loads. Any uncritical preference of the separate system as a particularly advantageous solution is thus questionable. Individual investigations case by case are recommended. PMID- 15790236 TI - Experimental analysis of the hydrass flushing gate and field validation of flush propagation modelling. AB - As well known, a periodical cleansing of sewers results necessary in order to avoid or reduce many unpleasant consequences due to the presence of sediments, such as the solid re-suspension during rain events and the reduction of the channel flow capacity. For this purpose, different systems based on flushing techniques have been adopted in sewers. In the present paper, the results of an analysis of the hydraulics of a flushing device, the Hydrass gate, is presented. In particular, an experimental investigation on a scale model under steady conditions was carried out in order to determine the behaviour of the device during the flushing phase. Outflow relations were derived for the different outflow conditions. A numerical model was finally set up for testing the relations under unsteady conditions, using for validation the experimental data of a previous measurement campaign carried out in a sewer reach of the city of Lyon, France. PMID- 15790237 TI - Inefficiency of storm water inlets as a source of urban floods. AB - The paper attempts to explain sources of surplus storm water runoff in urban areas, particularly in relation with functioning of inlets. Inlet capacity (quantity of captured water) and inlet efficiency (portion of the approaching flow rate) have been defined and their relationship with relevant parameters (approaching flow, longitudinal and lateral street slopes) was established through laboratory measurements. Effects of clogging of inlets on inlet capacity were also investigated in laboratory conditions. As a consequence of decreased inlet efficiency, there is a portion of approaching flow that is not captured by the inlet (pass-over flow). If the pass-over flow is considered along a street having numerous inlets, it is easy to estimate the quantity of flow that would accumulate on the pavement. Inlet inefficiency can be significant when overestimation of inlet capacity results in increased distance between consecutive inlets, and when clogging of grates or inadequate placing of inlets causes significant decrease in inlet capacity. PMID- 15790238 TI - Factors influencing exfiltration processes in sewers. AB - Exfiltration from sewers is widespread and emerging legislation may require water service providers to identify, and rectify, its sources in sewerage systems. This paper describes exfiltration test apparatus and a series of experiments undertaken using sewage to gain a better understanding of the influence of sewage solids and sediments on leakage rates. An overview of the results obtained is given, which demonstrates that most previous estimates of exfiltration leakage rates were too high due to a lack of appreciation of the "self-repairing" action of sewage and sewage associated solids. Exfiltration rates of 0.1% of the sewer flow or 0.001 I/s have been recorded for defects up to 6 mm wide. PMID- 15790239 TI - Sensitivity to experimental data of pollutant site mean concentration in stormwater runoff. AB - Urban wet weather discharges are known to be a great source of pollutants for receiving waters, which protection requires the estimation of long-term discharged pollutant loads. Pollutant loads can be estimated by multiplying a site mean concentration (SMC) by the total runoff volume during a given period of time. The estimation of the SMC value as a weighted mean value with event runoff volumes as weights is affected by uncertainties due to the variability of event mean concentrations and to the number of events used. This study carried out on 13 catchments gives orders of magnitude of these uncertainties and shows the limitations of usual practices using few measured events. The results obtained show that it is not possible to propose a standard minimal number of events to be measured on any catchment in order to evaluate the SMC value with a given uncertainty. PMID- 15790240 TI - A benchmark methodology for managing uncertainties in urban runoff quality models. AB - In this paper we present a benchmarking methodology, which aims at comparing urban runoff quality models, based on the Bayesian theory. After choosing the different configurations of models to be tested, this methodology uses the Metropolis algorithm, a general MCMC sampling method, to estimate the posterior distributions of the models' parameters. The analysis of these posterior distributions allows a quantitative assessment of the parameters' uncertainties and their interaction structure, and provides information about the sensitivity of the probability distribution of the model output to parameters. The effectiveness and efficiency of this methodology are illustrated in the context of 4 configurations of pollutants' accumulation/erosion models, tested on 4 street subcatchments. Calibration results demonstrate that the Metropolis algorithm produces reliable inferences of parameters thus, helping on the improvement of the mathematical concept of model equations. PMID- 15790241 TI - Test of the efficiency of three storm water quality models with a rich set of data. AB - The objective of this article is to test the efficiency of three different Storm Water Quality Model (SWQM) on the same data set (34 rain events, SS measurements) sampled on a 42 ha watershed in the center of Paris. The models have been calibrated at the scale of the rain event. Considering the mass of pollution calculated per event, the results on the models are satisfactory but that they are in the same order of magnitude as the simple hydraulic approach associated to a constant concentration. In a second time, the mass of pollutant at the outlet of the catchment at the global scale of the 34 events has been calculated. This approach shows that the simple hydraulic calculations gives better results than SWQM. Finally, the pollutographs are analysed, showing that storm water quality models are interesting tools to represent the shape of the pollutographs, and the dynamics of the phenomenon which can be useful in some projects for managers. PMID- 15790242 TI - CSOs: tools for assessing their operation in our systems. AB - In recent years, regulations have been introduced under European legislation to govern wastewater discharge into natural environments, with particular provisions for combined sewer overflows. It has therefore become indispensable to control the hydraulic behaviour of these constructions. This article addresses the issue of 1D and 3D hydraulic modelling of CSOs. In the case of 1D modelling, a computational tool that is adapted to the hydraulic complexity of side weirs has been designed. For 3D modelling, the Fluent calculation code has been tested. The two modelling approaches have been validated by the data collected from the small scale model in Obernai (French). PMID- 15790243 TI - Use of 3D modelling and several ultrasound sensors to assess overflow rate. AB - A new instrumentation mode has been put in place in order to determine the overflow rate of five complex CSOs of the system in Selestat (French), which are subject to self monitoring. 3D and 1D models have made it possible to predict the shapes of the water lines and suggest a location for the ultrasound sensors. In order to validate the instrumentation principle, three overhead sensors were placed on a weir. The overflow laws suggested are of the type Q(overflow) = a1h1b1 + a2h2b2 + a3h3b3. Early results of the overflow rate that have been obtained by applying the law are close to 10 % of the flow measured in the physical test bench. On the actual site, the ultrasound sensors were assembled on a rail covering the entire weir in order to be able to change their position in future after models calibration. PMID- 15790244 TI - Comparison of radar and raingauge measurements during heavy rainfall. AB - Five heavy small-scale rainfall events in North Rhine-Westphalia (Germany) were investigated with radar and raingauge data. Special attention was paid to quality check and adjustment of radar data. Attenuation effects could be observed on both, C-Band and on X-Band radar. Adjustment of radar data to raingauge values turned out to be very difficult in the vicinity of heavy local rain cells. For the five affected regions the precipitation was quantified in the form of areal time series and cumulated radar images. As further result of this project, the spatial extent of the precipitation fields was identified and compared with radar and raingauge data. PMID- 15790245 TI - Radar-based alert system to operate a sewerage network: relevance and operational effectiveness after several years of use. AB - Since January 2000, the sewerage network of a very urbanised catchment area in the Greater Nancy Urban Community has been operated according to the alarms generated in real time by a storm alert system using weather radar data. This alert system is based on an automatic identification of intense rain cells in the radar images. This paper presents the characteristics of this alert system and synthesises the main results of two complementary studies realised in 2002 in order to estimate the relevance and the operational effectiveness of the alert system. The first study consisted in an off-line analysis of almost 50,000 intense rain cells detected in four years of historical radar data. The second study was an analysis of the experience feedback after two years of operational use of this alert system. The results of these studies are discussed in function of the initial operational objectives. PMID- 15790246 TI - Development of the real-time control (RTC) system for Tokyo sewage system. AB - Tokyo Metropolitan government has decided to make the maximum possible use of the existing facilities while ensuring safety against inundation and to promote measures also from a software approach by introducing a system capable of minimizing combined sewer overflow, the real-time control system (RTC). A pilot RTC system was installed in August 2002 for the Shinozaki Pumping Station. The RTC system monitors the precipitation volume and the water level in the pipe. Simulations were carried out on the basis of these data. From the results, it was found that with the use of the RTC it is possible to reduce CSO by roughly 50% for small rainfalls with a total precipitation level of 20 mm or less by strong rainwater in the pipe routes at the beginning of the rain. It has also been shown that CSO can be reduced by about 80% through the use of rainfall forecasting. PMID- 15790247 TI - An integrated inundation model for highly developed urban areas. AB - A numerical model is developed in this study with various components for simulating the complex flow phenomena in urban drainage basins. The model integrates the HEC-1 model, a 1-D dynamic channel-flow model, a 2-D non-inertia overland-flow model and the SWMM model to reflect the hydraulic processes in areas with different characteristics. The inundation of underground infrastructure during flood is also considered in the model. The typhoon Nari event in 2001, which resulted in severe flood in downtown Taipei, is simulated by the model. The result is compared with the survey records of flooded areas, which reveals the storage effect of underground infrastrucures is significant to the simulation results of highly developed urban areas. PMID- 15790248 TI - Quantitative and qualitative hydrologic balance for a suburban watershed with a separate sewer system (Nantes, France). AB - A qualitative and quantitative budget at the outlet of the storm-water runoff system of a small suburban watershed is presented together with some data regarding waste-water. 445,000 m3 (34% of the rain-water volume) were drained by the storm-water runoff system and 40,879 m3 by the waste-water system from September 2002 to March 2004. Storm-water runoff is generally not heavily polluted with regard to trace metals but concentrations occasionally exceed the standards for surface water of good quality. On the contrary, pesticides (diuron and glyphosate) have very high concentrations especially in spring and autumn when their use is maximum. As the St Joseph storm-water runoff is finally discharged into the Erdre River, measures to reduce the use of these pollutants should be considered. PMID- 15790249 TI - Master planning for stream protection in urban watersheds. AB - Urbanization results in great changes to the landscape and the water environment simply because stormwater runoff differs in quantity and quality from the pre urbanization state. Streams, rivers, lakes, estuaries and other receiving water bodies experience the changes to runoff frequencies and volumes and react accordingly. The forces behind the observed changes in the receiving waters are discussed in this paper and suggestions are made on how to plan to deal with them. Urban watershed and waterway master planning can help to mitigate, in large part, the impacts imposed on these waters by land-use changes. Although each watershed is unique, some general principles are suggested to deal with these emergent problems. PMID- 15790250 TI - Identification of ecological status of stream impacted by urban drainage. AB - The adoption and implementation of the Directive 2000/60/EC with respect to crucial changes in an approach to water protection require a complex methodology for river and stream status assessment. This paper presents a possible method for assessing the ecological status of urban streams, on for example the Botic creek, the largest tributary of the Vltava River in Prague. The study stream does not possess a good ecological status. The degradation of the ecosystem quality is caused mainly by combined sewer overflows, which reduce water and sediment quality and induce chemical and hydraulic stress for the benthic community. PMID- 15790251 TI - Editorial--classification systems in clinical practice. PMID- 15790252 TI - Measurement of headache frequency, intensity and duration: comparison of patient report by questionnaire and headache diary. AB - BACKGROUND AND PURPOSE: The efficacy of interventions for headache is often based on patient estimates of the headache parameters of frequency, intensity and duration, which can be ascertained from interview, by questionnaire or from a headache diary. The aim of the present study was to test the accuracy of retrospective patient reports of headache frequency, intensity and duration, with respect to measures derived from a daily headache diary. METHOD: Forty participants suffering from least one headache per month completed a daily headache diary for four weeks. At the end of the four-week period participants completed a questionnaire requiring them to estimate the frequency, usual duration and usual intensity of their headaches. Spearman rank correlation coefficients were calculated to determine the relationship between diary and questionnaire data. RESULTS: Correlation coefficients (Spearman's rho) for questionnaire and diary data for headache frequency and duration were 0.80 and 0.72, respectively, whereas that for intensity was 0.51. CONCLUSIONS: The findings suggest that when compared with the use of a headache diary, patient estimations of headache frequency and duration by questionnaire are reasonably accurate. Headache intensity appears to be more difficult to remember and report, possibly because of the multidimensional nature of pain, as opposed to the temporal characteristics of frequency and duration. It is recommended that frequency and duration be preferred over intensity as outcome measures if questionnaires or retrospective reports are used. PMID- 15790253 TI - Obstetric brachial plexus palsy: increasing disability and exacerbation of symptoms with age. AB - BACKGROUND: People with congenital conditions such as obstetric brachial plexus palsy (OBPP), sometimes known as Erb's palsy, may experience adverse effects as a result of compensatory strategies adopted to achieve improved function. Anecdotal evidence suggests that problems do occur in adult life but little is known about the type and extent of problems experienced, nor whether they are similar to those with other conditions involving movement dysfunction. METHOD: A survey of adults with OBPP to determine problems currently experienced. Subjects were members of the Erb s Palsy Group UK. RESULTS: Forms were sent to all adult members of the group (n = 61), and replies were received from 36 (59%). The majority of the sample reported many problems, including pain, impaired sensation, arthritis and limitation of function, which interfered with activities of daily living. CONCLUSION: In the present small sample, most adults with OBPP reported experiencing many difficulties, some similar to those experienced by others with movement dysfunction. The topic warrants further investigation, both in terms of avoiding these problems where possible and, when complications occur, considering how they can best be managed. PMID- 15790254 TI - Physiotherapy and spinal nerve root adhesion: a caution. AB - BACKGROUND AND PURPOSE: The term 'spinal neuropathic pain' has been coined to describe the chronic neuropathic pain that results when spinal nerve roots are aggravated by scar tissue. (It is different from the pain of spinal cord injury.) Such patients have longstanding back and radicular pain (nerve root pain, predominantly in the limbs) caused by scar or inflammatory tissue around the nerve roots. The pathology of such patients' pain means that special consideration needs to be given to the fact that such adhesions compromise nerve biomechanics and that movement generates additional pain. Patients with such spinal neuropathic pain often do not do well from conventional physiotherapy. Exacerbation (flare-up) frequently follows the exercise routines in common practice. METHOD: Individual patient experience was collected from an internet support group, and the results were tabulated. RESULTS: All patients considered stretching, flexing and strenuous exercise to be harmful. A few reported that gentle exercise with instruction not to cause pain was beneficial. Some patients received advice not to do physiotherapy once they had been diagnosed with arachnoiditis. CONCLUSIONS: The treatment of patients with spinal neuropathic pain warrants special consideration as far as physiotherapy is concerned: patients should only be prescribed gentle, individually tailored exercise. It is hoped the present small study will promote understanding and the development of better therapy. PMID- 15790255 TI - Physiotherapy management of patients with HIV-associated Kaposi's sarcoma. AB - BACKGROUND: Kaposi's sarcoma is the most common form of cancer in patients with human immunodeficiency virus (HIV) infection. Although Kaposi sarcoma lesions may contribute to significant physical impairments, there is a lack of scientific literature detailing the role of physiotherapy in the treatment of HIV-associated Kaposi's sarcoma. The present Case Report includes two males, aged 36 and 39 years, seropositive for HIV with invasive Kaposi's sarcoma. METHOD AND RESULTS: Patient A was evaluated for bilateral foot pain caused by plantar surface Kaposi s sarcoma lesions that rendered him unable to walk. He progressed to walking 400feet after a treatment regimen of gait training with the use of custom plastazote sandals. Patient B was evaluated for right lower extremity lymphoedema secondary to invasive Kaposi's sarcoma. He experienced an 18% reduction in limb volume, a 38% reduction in pain and a 20 degrees increase in terminal knee flexion after therapeutic exercise and the use of compressive bandaging and garments. CONCLUSIONS: This Case Report suggests that physiotherapy interventions may be valuable in the conservative management of patients with HIV-associated Kaposi s sarcoma. PMID- 15790256 TI - Why is the functional independence measure used to identify some rehabilitation needs in stroke survivors when there are better tools? PMID- 15790258 TI - [What policy for the family in 2005?]. PMID- 15790260 TI - [Migraines in children. A frequent but little known pathology]. PMID- 15790261 TI - [Epidemiology and diagnosis of migraines in children]. PMID- 15790262 TI - [Therapeutic management of migraines in children]. PMID- 15790263 TI - [Relaxation groups for children with migraines]. PMID- 15790264 TI - [Hypnosis and migraines in children]. PMID- 15790265 TI - [Nothing explained why Melanie suffered from "headaches"]. PMID- 15790266 TI - [Julie's migraine was diagnosed by an orthodontist]. PMID- 15790267 TI - [Mother's milk, sacred food, power of life, power of death]. PMID- 15790268 TI - [Pediatric nursing in a medical-forensic unit]. PMID- 15790269 TI - [3/6 Use of equimolar mix of oxygen and nitrogen monoxide (Meopa)]. PMID- 15790270 TI - [SPARADRAP]. PMID- 15790271 TI - Continuing to support provision of safe care. PMID- 15790272 TI - Award winning website. PMID- 15790273 TI - Calling future nurse practitioners! PMID- 15790274 TI - Understanding pain management. PMID- 15790275 TI - Gerontological nurses group aims to educate. PMID- 15790276 TI - Born leaders. PMID- 15790277 TI - Collaborating for healthy campuses. PMID- 15790278 TI - Liability insurance coverage when working in a private medical clinic. PMID- 15790279 TI - Nursing leadership. PMID- 15790280 TI - Caring for patients with heart failure. AB - Heart failure affects more: 350,000 Canadians and accounts for more than dollar 1 billion of Canada's health budget. It is the most common condition requiring re hospitalization and carries with it a mortality rate of between 25% and 40% (Liu et al., 2003) In British Columbia, approximately 70,000 people (1.5% of the B.C. population) were diagnosed with heart failure in 2003 and it is estimated that a further 70,000 people may be living undiagnosed with the condition (British Columbia Ministry of Health Services, 2003). The vast majority of people with heart failure are over 70 years of age. As our population ages, it is reasonable to expect that the burden of heart failure on our health system will continue to increase. Heart failure is usually a chronic condition marked by frequent exacerbations. It is these exacerbations that bring patients to hospital emergency departments and medical wards. PMID- 15790281 TI - Regulations and scope of practice. PMID- 15790282 TI - Transition costs included in budget. PMID- 15790284 TI - Patient safety and duty to report. PMID- 15790283 TI - Used fentanyl patch could lead to an inadvertent overdose. PMID- 15790285 TI - Planes, trains & automobiles. PMID- 15790286 TI - Nursing communication. PMID- 15790287 TI - Taking time to do an RFP the right way. More than you bargained for. PMID- 15790288 TI - Net worth. Intranet ordering system proves successful. PMID- 15790289 TI - New polymer materials in total hip arthroplasty. Evaluation with radiostereometry, bone densitometry, radiography and clinical parameters. AB - AIMS: To evaluate the outcome of different types of polyethylene, bone cements and one design of uncemented fixation with porous and ceramic coating using radiostereometry, bone densitometry, conventional radiography and clinical parameters. MATERIALS AND METHODS: Study I: 201 patients were extracted from 5 prospective randomised studies to evaluate femoral head penetration at two years with radiostereometry in four basic designs, cemented Lubinus and Reflection cups, uncemented Trilogy and Reflection cups. Studies II and III: 60 patients (61 hips) were randomised to receive either highly cross-linked or conventional all PE cups. 32 patients with bilateral arthrosis received hybrid THA with highly cross-linked PE on one side and conventional on the contra lateral side. Femoral head penetration and the migration of the cups were evaluated with radiostereometry in the supine and standing positions. DEXA and conventional radiography were used to evaluate the bone mineral density and radiolucencies around the cemented acetabular component. Studies IV and V: 90 patients (97 and 96 hips respectively) were stratified depended on age, gender, diagnosis and preoperative BMD to create 3 main groups of socket fixation. In the first group fluoride containing cement was used, in the second group Palacos cum Gentamicin and in the third hybrid THA with porous coated HA/TCP cup. In the hybrid group the fixation of the femoral component was again randomised to either of the two cements. The results on femoral and acetabular sides are presented separately in studies IV and V, respectively. RESULTS: Study I: Cups with polyethylene sterilized in EtO had almost twice the proximal and 3D penetration rates compared with gamma-sterilized polyethylene. Regression analysis showed that the type of sterilization, age and weight was the most important factors affecting the penetration rate. Studies II and III: In the cemented study the proximal penetration was lower in the study group independent of position at 3 years, while in the hybrid study the penetration was lower in the study group only in the supine position at 2 years. The migration of the cup did not differ between the plastics in both studies. At 2 years the periprosthetic radiolucency and BMD did not differ significantly between the 2 types of PE used in the cemented study. Study IV: The subsidence of the stem did not differ between the groups, but the periprosthetic BMD decreased more in fluoride cement group at 2 years. Conventional radiography revealed higher progression of radiolucent lines in the Palacos group, but only in one region. Study V: The proximal migration of the cup was almost similar in all three groups. The three dimensional migration was increased in patients with osteoporosis. Postoperative radiolucent lines tended to disappear with use of porous coating covered with HA/TCP. CONCLUSIONS: Study I: EtO sterilized polyethylene increased the femoral head penetration. Age and weight were also important predictors of the penetration rate. Studies II and III: The highly cross-linked polyethylene decreased the penetration rate mainly after one year probably reflecting less wear. The different mechanical properties of the two types of PE studied did not affect the early fixation of the cemented cup. Study IV: There were no obvious advantage of addition of fluoride to acrylic bone cement when used to fixate the femoral component. Study V: Use of fluoride containing cement or uncemented fixation did not improve the early stability of the socket compared to Palacos with Gentamicin. PMID- 15790290 TI - Infections in the elderly. PMID- 15790291 TI - Zentralblatt fur Bakteriologie--100 years ago: bacteriological warfare against mice and rats. PMID- 15790292 TI - Genetics of glycopeptide resistance in gram-positive pathogens. AB - Glycopeptide resistance in enterococci, first detected in 1986, has since spread worldwide and represents a major health problem. Importantly, it has been recently acquired by Staphylococcus aureus. We shall review the genetics which accounts for this successful dissemination and discuss the possible origins of the resistance genes. PMID- 15790293 TI - Hierarchical gene regulators adapt Salmonella enterica to its host milieus. AB - The facultative intracellular pathogen Salmonella enterica serovar Typhimurium possesses an elaborate set of virulence genes that enables the bacterium successfully to move between and adapt to the environment, different host organisms and various micro-niches within a given host. Expression of virulence attributes is by no means constitutive. Rather, the regulation of virulence determinants is highly coordinated and integrated into normal bacterial physiological responses. By integrating discriminating virulence gene regulators with conserved housekeeping regulatory processes, the bacteria can sense alterations in the repertoire of environmental cues, and translate the sensing events into a pragmatic and coordinated expression of virulence genes. While the description of transmissible genetic elements that import global gene regulatory factors into a cell brings conceptual problems into the established regulatory network, the existence of mobile gene regulators may actually enable the bacteria to further modulate virulence expression. PMID- 15790294 TI - Molecular evolution of the lysine decarboxylase-defective phenotype in Shigella sonnei. AB - Shigella, the etiological agent of the bacillary dysentery, belongs to the extremely diverse species of Escherichia coli. In the evolutionary route of Shigella from commensal E. coli ancestors towards a pathogenic lifestyle, the critical events have been the acquisition of the pINV plasmid, through horizontal transfer and the inactivation of pre-existing genes. These so-called pathoadaptive mutations affect the expression of genes negatively interfering with the newly acquired functions necessary for the colonization of the host niche. Cadaverine, a small polyamine resulting from decarboxylation of lysine, has been shown to hamper the full expression of Shigella invasiveness mainly by altering the inflammatory response. Recent analysis of the evolution of the Shigella and enteroinvasive E. coli (EIEC) cad region indicates that silencing of the cad locus has been attained with several strategies. The increasing relevance of S. sonnei in both, developing and industrial countries, prompted us to analyze the molecular origin of the LDC- phenotype in these strains. The results obtained on several S. sonnei strains reveal that despite the difference in geographic origin and antibiotic resistance patterns, all the strains have undergone the same modifications. Multiple IS insertions into the cadBA operon have interrupted gene continuity without inducing deletions or inversions of the cadA and cadB genes which are remained entirely conserved. Moreover, by functional analysis we show that all the strains carry a defective cadC gene, thus strengthening the hypothesis that inactivation of the regulatory cadC gene might have been the first step towards a complete lack of the cad locus. PMID- 15790295 TI - Occurrence of insertion sequences within the genomes and Tn1546-like elements of glycopeptide-resistant enterococci isolated in Brazil, and identification of a novel element, ISEfa5. AB - Insertion sequences (IS) occur widely within the Tn1546-like elements responsible for VanA glycopeptide resistance in enterococci from several countries. As such insertions can be used as epidemiological markers and for studying horizontal transfer of gene clusters, we investigated the distribution of IS6770, IS1542, IS1216V, IS1476, and IS1251 elements in 26 VanA Enterococcus faecium and 21 VanA Enterococcus faecalis from Brazil. PCR, using genomic DNA as a template, indicated that most of the isolates contained IS6770 (97%), IS1216V (87%) and IS1476 (72%) elements. IS1251 was also detected, but at a higher frequency in E. faecium (80%) than in E. faecalis (14%). None of the isolates harboured IS1542. Only two of 47 isolates had IS elements within their Tn1546-like elements; one possessed IS1251 between vanS and vanH, as reported in the United States; another possessed a novel IS element, designated ISEfa5, located between vanX and van Y. This novel element was found in the genomic DNA of 25 (96%) E. faecium and II (52%) E. faecalis. In stability studies, no IS-mediated changes were detected in the Tn1546-like elements of 25 vancomycin-resistant enterococci (VRE) monitored over 11 months. These results suggest that the occurrence of IS in Brazilian isolates is similar to that reported in American isolates, but that these elements occur rarely within the vanA gene clusters. As patterns of IS carriage did not correlate with the PFGE type of the VRE, the prevalence of IS elements in genomic DNA of VRE is not a useful epidemiological marker. However, the presence of IS-modified Tn1546-like elements, which appear to be rare in Brazil, could be a useful molecular marker in local epidemiological studies to monitor the evolution and horizontal transmission of VanA elements. PMID- 15790296 TI - Long-term clonal lineages within Campylobacter jejuni O:2 strains from different geographical regions and hosts. AB - The genomic diversity of 11 epidemiologically unrelated strains of Campylobacter jejuni (serotype O:2) isolated from different hosts and different geographical regions in Germany over a period of 15 years was studied and results were compared with the reference strain NCTC11168. By flagellin PCR-RFLP typing six fla types were identified, while macrorestriction analysis with three different restriction enzymes revealed almost identical patterns for two human and one bovine strain, even though they were isolated between 1984 and 1996. Interestingly, the PFGE and fla profiles from strain NCTC11168, which was originally isolated in 1977 from an outbreak case in Worcester (UK), were highly similar or even identical to the profiles of these strains. Besides this, mapping of selective genetic markers to the obtained restriction fragments by Southern blot hybridization showed a conserved localization of the investigated sequences in several strains. Our data confirmed considerable degrees of genomic conservation and the occurrence of long-term O:2 serotype-associated clonal lineages in C. jejuni in different geographical regions and hosts. PMID- 15790297 TI - DNA sequence-based tandem repeat analysis of the clfB gene is less discriminatory than spa typing for methicillin-resistant Staphylococcus aureus. AB - Methicillin-resistant Staphylococcus aureus (MRSA) are a major challenge for hospital hygiene. Typing by DNA sequence analysis of the repeat region of the protein A gene (spa) significantly improved typing of MRSA in the hospital setting. However, microevolution of spa repeats in epidemic clones appears to occur at a fairly slow clock rate. Therefore, DNA sequence-based methods providing additional resolution are desirable in some situations. We evaluated the use of the clfB repeat region proposed recently by others as a possible complementation to spa typing. Using epidemic MRSA isolates from two German university hospitals, we show that the clfB repeat region does not offer any additional discriminatory power. PMID- 15790298 TI - Intraocular microsporidiosis due to Encephalitozoon cuniculi in a patient with idiopathic CD4+ T-lymphocytopenia. AB - Encephalitozoon cuniculi was documented to cause disseminated microsporidial infection including an iris tumor and endophthalmitis in an adolescent with idiopathic CD4+ T-lymphocytopenia. The diagnosis was established by microscopic, serologic and molecular methods. E. cuniculi (rabbit strain) was identified from the iris tumor, as well as from urine, conjunctival, corneal, and nasal swabs. Treatment with oral albendazole led to rapid improvement. This case raises the possibility of disseminated microsporidial infection in the context of idiopathic CD4+ T-lymphocytopenia and possibly advanced human immunodeficiency virus (HIV) infection, and above all the possibility of intraocular infection with E. cuniculi in humans. PMID- 15790299 TI - Summary health statistics for U.S. adults: National Health Interview Survey, 1998. AB - OBJECTIVES: This report presents health statistics from the 1998 National Health Interview Survey for the civilian noninstitutionalized adult population, classified by sex, age, race and Hispanic origin, poverty status, and region of residence for chronic condition prevalence, health status and limitations in activity, health care access and utilization, health behaviors, and attitudes toward acquired immunodeficiency syndrome (AIDS). Health statistics by education, income, health insurance coverage, marital status, and place of residence are also presented for health status and limitations in activity, health care access and utilization, health behaviors, and knowledge and attitudes toward AIDS. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized U.S. population. Data are collected during face-to-face interviews with adults present at the time of the interview. Also, the self-reported health information for adults in this report was obtained from one randomly selected adult per family. HIGHLIGHTS: In 1998, 65% of adults 18 years of age and over reported excellent or very good health. However, 17% of adults less than 65 years of age had no health insurance coverage, and 14% of adults did not have a usual place of medical care. Twelve percent of adults had ever been told by a doctor or health professional that they had heart disease and 19% had been told on two or more visits that they had hypertension. Nearly a quarter of adults were current smokers, and 23% were former smokers. Sixty percent did not engage in any leisure-time vigorous physical activity, and based on estimates of body mass index, 35% were overweight and 20% were obese. PMID- 15790300 TI - Drug delivery of aminolevulinic acid from topical formulations intended for photodynamic therapy. AB - Topical photodynamic therapy is used for a variety of malignant and pre-malignant skin disorders, including Bowen's Disease and Superficial Basal Cell Carcinoma. A haem precursor, typically 5-aminolevulinic acid (ALA), acting as a prodrug, is absorbed and converted by the haem biosynthetic pathway to photoactive protoprophyrin IX (PpIX), which accumulates preferentially in rapidly dividing cells. Cell destruction occurs when PpIX is activated by an intense light source of appropriate wavelength. Topical delivery of ALA avoids the prolonged photosensitivity reactions associated with systemic administration of photosensitisers but its clinical utility is influenced by the tissue penetration characteristics of the drug, its ease of application and the stability of the active agent in the applied dose. This review, therefore, focuses on drug delivery applications for topical, ALA-based PDT. Issues considered in detail include physical and chemical enhancement strategies for tissue penetration of ALA and subsequent intracellular accumulation of PpIX, together with formulation strategies and drug delivery design solutions appropriate to various clinical applications. The fundamental aspects of drug diffusion in relation to the physicochemical properties of ALA are reviewed and specific consideration is given to the degradation pathways of ALA in formulated systems that, in turn, influence the design of stable topical formulations. PMID- 15790301 TI - Physical and chemical characterization of iris and choroid melanosomes isolated from newborn and mature cows. AB - Bovine iris and choroid melanosomes at two ages (<1 week and >2 years) were examined by inductively coupled plasma mass spectrometry (ICP-MS), elemental analysis, infrared spectrometry (IR) and X-ray photoelectron spectrometry (XPS). When iris and choroid melanosomes at the same age were compared, the quantification of metal elements by ICP-MS revealed that choroid melanosomes had a higher binding capacity for the carboxylate-binding metal ions (e.g. Na+ K+, Mg2+, Ca2+ and Zn2+). Elemental analysis showed a higher O:N ratio in choroid melanosomes. Both observations suggested that choroid melanosomes have a higher content of carboxylate-containing monomer than iris melanosomes. IR spectrometric analysis showed a red shift (approximately 8 cm(-1)) of the absorption peak of aromatic C=C, C=N and C=O at approximately 1630 cm(-1) in the IR spectrum of iris melanosomes relative to choroid melanosomes. Increased conjugation in the molecular structure of the pigment is proposed to contribute to this peak shift. It is also notable that although the elemental analysis showed different C, N and O contents in the two types of melanosomes, XPS showed almost the same elemental compositions on the surface of two types of iris and choroid melanosomes studied. When the melanosomes from the same tissues at different ages were compared, ICP MS analysis suggested that the number of carboxylate groups in the melanosomes decreased with age. Both elemental analysis and XPS showed that C:N ratio decreased with age, which was proposed to be due to both a decrease in carboxylate groups in mature samples and to the fissure of phenol rings caused by age-associated oxidation. Such age-related oxidative damage diminishes conjugation and is manifested by blue shifts of absorption peaks for aromatic double bonds in the IR spectra of mature melanosomes. XPS analysis showed that the ratio of C-O:C=O decreased with age. These tissue-related and age-related chemical differences between samples affected the optic density and metal binding properties of melanosomes, which are believed to be closely associated with the biological functions of melanins. PMID- 15790302 TI - Rational Peptide-based vaccine design for cancer immunotherapeutic applications. AB - Immune responses to cancer cells can be elicited in vivo by administering synthetic peptides derived from proteins uniquely or overexpressed on tumor cells (tumor associated antigens--TAAs). Peptides derived from TAAs are presented in the context of major histocompatibility complex (MHC) molecules to cytotoxic T cells (CTL), which can recognize and lyze tumor cells. In contrast to peptides derived from an exogenous source (viral or bacterial), tumor peptides bind weakly to MHC class I molecules. The low binding affinity of these peptides makes them poor candidates for vaccination due to the poor immunogenic response produced. In order to enhance antigen recognition and hence immunogenicity, peptide binding affinity for MHC can be initially improved by modifying the "anchor" residues. However, the task at hand is highly unpredictable and minor changes in peptide sequence can alter/abolish the T cell response. Furthermore, despite the wealth of information obtained over the last decade from high resolution X-ray structures of MHC class I in complex with peptides (pMHC) as well as pMHC in complex with T cell receptor (TcR), prediction remains difficult. Nonetheless, peptides represent convenient chemical entities that can be rapidly synthesized in clinical grade for therapeutic applications. Herein, the rationale behind modifying TAAs will be discussed including the synthesis/use of proteolytically tolerant peptides (and peptide mimetics) which incorporate non-natural amino acids, retro-inversion and cyclization to improve bioavailability. PMID- 15790303 TI - Photoaffinity labeling technique for studying DNA replication and DNA repair. AB - The photoaffinity labeling technique is based on UV-light induced crosslinking of proteins to photoreactive DNA. Photoreactive groups can be introduced at different points of DNA (base or phosphate) either at the ends or in inner positions of the DNA chain by combination of enzymatic and chemical synthesis. Varying the structure of photoreactive DNA one can design the intermediates of different stages of DNA replication or DNA repair and apply them to identify the proteins crosslinked to specific positions of the DNA chain in single- or double stranded DNA, partial DNA duplexes, gapped or nicked duplexes or DNA carrying flap structures. A wide range of base-substituted dNTP analogs containing photoreactive groups of different photoreactivity and spacers of various lengths has been synthesized and characterized. Photoreactive dNTP analogs have been shown to be effective substrates of viral, bacterial and eukaryotic DNA polymerases. The efficiencies of crosslinking of DNA bearing various photoreactive dNMP to protein and DNA targets were estimated. New approaches for enzymatic introduction of photoreactive groups onto the 5'-end of oligonucleotides and into the inner positions of DNA chain have been elaborated. Photoreactive DNAs have been successfully used to study enzymes and protein factors of DNA replication and repair in reconstituted systems and cellular/nuclear extracts. Photoaffinity labeling technique was shown to be a prominent tool of proteomics to elucidate structural and functional aspects of protein-DNA interactions. It can be also applied for identification of proteins including unknown ones, which interact with specific DNA intermediates in cellular/nuclear extracts. PMID- 15790305 TI - The medicinal chemistry of botulinum, ricin and anthrax toxins. AB - The potential use of weapons of mass destruction (nuclear, biological or chemical) by terrorist organizations represents a major threat to world peace and safety. Only a limited number of vaccines are available to protect the general population from the medical consequences of these weapons. In addition there are major health concerns associated with a pre-exposure mass vaccination of the general population. To reduce or eliminate the impact of these terrible threats, new drugs must be developed to safely treat individuals exposed to these agents. A review of all therapeutic agents under development for the treatment of the illnesses and injuries that result from exposure to nuclear, biological or chemical warfare agents is beyond the scope of any single article. The intent here is to provide a focused review for medicinal and organic chemists of three widely discussed and easily deployed biological warfare agents, botulinum neurotoxin and ricin toxins and the bacteria Bacillus anthracis. Anthrax will be addressed because of its similarity in both structure and mechanism of catalytic activity with botulinum toxin. The common feature of these three agents is that they exhibit their biological activity via toxin enzymatic hydrolysis of a specific bond in their respective substrate molecules. A brief introduction to the history of each of the biological warfare agents is presented followed by a discussion on the mechanisms of action of each at the molecular level, and a review of current potential inhibitors under investigation. PMID- 15790304 TI - Betulinic acid and its derivatives: a review on their biological properties. AB - Betulinic acid is a naturally occurring pentacyclic triterpenoid and has been shown to exhibit a variety of biological activities including inhibition of human immunodeficiency virus (HIV), antibacterial, antimalarial, antiinflammatory, anthelmintic and antioxidant properties. This article reports a survey of the literature dealing with betulinic acid related biological properties that has appeared from the 1990's to the beginning of 2003. A broad range of medical and pharmaceutical disciplines are covered, including a brief introduction about discovery, phytochemical aspects, organic synthesis, anti-HIV and cytotoxic mechanisms of action. Various structural modifications carried out and their biological and pharmacokinetic profiles are also incorporated. PMID- 15790306 TI - Transcription factor therapeutics: long-shot or lodestone. AB - Many drugs, both approved and in development, exert their effects through transcription factors. The complexity of the biology of transcriptional regulation, however, presents challenges to the effective design of therapies that directly target transcription factors. This review focuses on the different approaches that are currently pursued in therapeutics and drug discovery aimed at targeting transcription factors and signaling molecules, and their areas of effect in transcription factor biology. A relatively recent approach termed "transcription therapy" has been applied for a number of drugs in development, which may potentially provide a new avenue for targeting transcription factors for therapeutic purposes. PMID- 15790307 TI - Combinatorial natural products: from cloning to analysis. AB - Medicinal compounds from plants represent one of the largest and most diverse groups of plant secondary metabolites. The advent of advanced bioinformatics tools and modern genetic technology allowed for manipulation of biosynthetic pathways with the potential of generating novel chemical entities. First, public databases of secondary metabolite related enzymes were interrogated to identify relevant plant genes from vinca rosea (Catharanthus roseus) and other species. Genes of interest were tested after cloning by transfection into tobacco cell cultures using DNA viral vectors. The biosynthetic enzymes coded by these genes were over-expressed in the host. Automated solvent extraction procedure was employed to extract secondary metabolites from plant leaf tissues and transfected tobacco cell culture samples. The composition of the extracts was analyzed by state of the art bioanalytical methods such as high performance liquid chromatography and capillary electrophoresis to monitor changes in secondary metabolite patterns. PMID- 15790308 TI - Prenylated flavonoids: pharmacology and biotechnology. AB - Within the flavonoid class of natural products the prenylated sub-class is quite rich in structural variety and pharmacological activity. In the last twenty years a huge number of new structures has been reported, mostly from Leguminosae and Moraceae, with few coming from other genera. The presence, in different forms, of the isoprenoid chain can lead to impressive changes in biological activity, mostly attributed to an increased affinity for biological membranes and to an improved interaction with proteins. Molecules, such as xanthohumol and sophoraflavanone G, while being very structurally simple, show numerous pharmacological applications and are ideal candidates for SAR aimed to the discovery of new drugs. Only recently the biogenesis of these compounds has been more extensively studied and much attention has been focused on the enzymes involved in the modification and transfer of the prenyl unit. PMID- 15790309 TI - Regulatory and accessory HIV-1 proteins: potential targets for HIV-1 vaccines? AB - The HIV-1 regulatory proteins Tat and Rev and the accessory proteins Vpr, Vpu and Vif are essential for efficient viral replication, and their cytoplasmic production suggests that they should be processed for recognition by cytotoxic T lymphocytes. However, only limited data is available, evaluating the role of immune responses directed against these proteins in natural HIV-1 infection. Recent advances in the methods used for the characterization of HIV-1-specific cellular immune responses, including quantification of antigen-specific IFN-gamma production by ELISpot assay and flow-cytometry-based intracellular cytokine quantification, have allowed for a much more comprehensive assessment of virus specific immune responses. Emerging data show that the regulatory and accessory proteins serve as important targets for HIV-1-specific T cell responses, and multiple CTL epitopes have been identified in functionally important regions of these proteins. Moreover, the use of autologous peptides have allowed for the detection of significantly stronger HIV-1-specific T cell responses in the more variable regulatory and accessory HIV-1 proteins Tat and Vpr. These data indicate that despite the small size of these proteins, regulatory and accessory proteins are targeted by cellular immune responses in natural HIV-1 infection and contribute importantly to the total HIV-1-specific CD8+ T cell response. A multi component vaccine, with the inclusion of these proteins plus structural proteins remains the most promising choice for an effective AIDS vaccine. PMID- 15790310 TI - Physical and functional interactions between members of the tumour suppressor p53 and the Sp families of transcription factors: importance for the regulation of genes involved in cell-cycle arrest and apoptosis. AB - In the present study, we have investigated mechanisms of transcriptional co operation between proteins that belong to the tumour suppressor p53 and Sp (specificity protein) families of transcription factors. Such mechanisms may play an important role in the regulation of genes containing binding sites for both classes of transcription factors in their promoters. Two of these genes were analysed in the present study: the cyclin-dependent kinase inhibitor p21Cip1 gene and the PUMA (p53-up-regulated mediator of apoptosis) gene. We found that Sp1 and Sp3, but not Sp2, co-operate functionally with p53, p73 and p63 for the synergistic transactivation of the p21Cip1 promoter in Drosophila Schneider SL2 cells that lack endogenous Sp factors. We also found that Sp1 strongly transactivated the PUMA promoter synergistically with p53, whereas deletion of the Sp1-binding sites abolished the transactivation by p53. Using p53 mutant forms in GST (glutathione S-transferase) pull-down assays, we found that the C terminal 101 amino acids of p53, which include the oligomerization and regulatory domains of the protein, are required for the physical interactions with Sp1 and Sp3, and that deletion of this region abolished transactivation of the p21Cip1 promoter. Utilizing truncated forms of Sp1, we established that p53 interacted with the two transactivation domains A and B, as well as the DNA-binding domain. Our findings suggest that Sp factors are essential for the cellular responses to p53 activation by genotoxic stress. Understanding in detail how members of the p53 and Sp families of transcription factors interact and work together in the p53-mediated cellular responses may open new horizons in cancer chemotherapy. PMID- 15790312 TI - Fibulin-5 interacts with fibrillin-1 molecules and microfibrils. AB - Fibulin-5 plays an important role in elastic fibre formation in vivo. We have investigated the molecular interactions between fibulin-5 and components of fibrillin-rich microfibrils which form a template for elastin. Fibulin-5 interacted in a dose-dependent manner with a fibrillin-1 N-terminal sequence and with tropoelastin, but not with MAGP-1 (microfibril-associated glycoprotein-1) or decorin. Fibulin-5 did not inhibit interactions between fibrillin-1 N- and C terminal fragments, or fibrillin-1 interactions with tropoelastin. Fibulin-5 may provide a link between tropoelastin and microfibrils in the pericellular space during elastic fibre assembly. PMID- 15790311 TI - Protein tyrosine phosphatase hPTPN20a is targeted to sites of actin polymerization. AB - The human genome encodes 38 classical tyrosine-specific PTPs (protein tyrosine phosphatases). Many PTPs have been shown to regulate fundamental cellular processes and several are mutated in human diseases. We report that the product of the PTPN20 gene at the chromosome locus 10q11.2 is alternatively spliced to generate 16 possible variants of the classical human non-transmembrane PTP 20 (hPTPN20). One of these variants, hPTPN20a, was expressed in a wide range of both normal and transformed cell lines. The catalytic domain of hPTPN20 exhibited catalytic activity towards tyrosyl phosphorylated substrates, confirming that it is a bona fide PTP. In serum-starved COS1 cells, hPTPN20a was targeted to the nucleus and the microtubule network, colocalizing with the microtubule-organizing centre and intracellular membrane compartments, including the endoplasmic reticulum and the Golgi apparatus. Stimulation of cells with epidermal growth factor, osmotic shock, pervanadate, or integrin ligation targeted hPTPN20a to actin-rich structures that included membrane ruffles. The present study identifies hPTPN20a as a novel and widely expressed phosphatase with a dynamic subcellular distribution that is targeted to sites of actin polymerization. PMID- 15790313 TI - Deletions within a CA-repeat-rich region of intron 4 of the human SP-B gene affect mRNA splicing. AB - Length variants within a CA-repeat-rich region of intron 4 of the human SP-B (pulmonary surfactant protein-B) gene are associated with several lung diseases. The hypothesis that SP-B intron 4 affects mRNA splicing was studied. SP-B minigenes containing exons 1-6 with a normal-sized intron 4 (pBi4normal) or intron 4 containing deletions (pBi4del) of 193, 211, 264 or 340 bp were expressed in CHO (Chinese hamster ovary) cells by transient transfection. Two forms of SP-B transcripts, normal and incompletely spliced, were detected. With pBi4normal, normal-sized SP-B mRNA was the predominant form and a very low amount of incompletely spliced mRNA was present, whereas with the pBi4del variants the amount of normal SP-B mRNAs was lower and the amount of incompletely spliced mRNA was relatively high. Reverse transcription-PCR results and sequencing data indicated that the incompletely spliced SP-B RNA contained intron 4 sequence, and this incompletely spliced RNA was also observed in normal lung. Lung cancer tissues with intron 4 deletions exhibited a larger amount of abnormally spliced RNAs compared with normal lung tissue or cancerous tissue with normal-sized intron 4. The results indicate that intron 4 length variants affect SP-B mRNA splicing, and that this may contribute to lung disease. PMID- 15790314 TI - Structural basis for the interaction between human milk oligosaccharides and the bacterial lectin PA-IIL of Pseudomonas aeruginosa. AB - One of the mechanisms contributing to the protection by breast-feeding of the newborn against enteric diseases is related to the ability of human milk oligosaccharides to prevent the attachment of pathogenic bacteria to the duodenual epithelium. Indeed, a variety of fucosylated oligosaccharides, specific to human milk, form part of the innate immune system. In the present study, we demonstrate the specific blocking of PA-IIL, a fucose-binding lectin of the human pathogen Pseudomonas aeruginosa, by milk oligosaccharides. Two fucosylated epitopes, Lewis a and 3-fucosyl-lactose (Lewis x glucose analogue) bind to the lectin with dissociation constants of 2.2x10(-7) M and 3.6x10(-7) M respectively. Thermodynamic studies indicate that these interactions are dominated by enthalpy. The entropy contribution is slightly favourable when binding to fucose and to the highest-affinity ligand, Lewis a. The high-resolution X-ray structures of two complexes of PA-IIL with milk oligosaccharides allow the precise determination of the conformation of a trisaccharide and a pentasaccharide. The different types of interaction between the oligosaccharides and the protein involve not only hydrogen bonding, but also calcium- and water-bridged contacts, allowing a rationalization of the thermodynamic data. This study provides important structural information about compounds that could be of general application in new therapeutic strategies against bacterial infections. PMID- 15790316 TI - The diagnosis and management of Kawasaki disease. AB - Kawasaki disease (KD) is a systemic vasculitis of childhood with a predilection for the coronary arteries. It is the predominant cause of paediatric acquired heart disease in developed countries. The aetiology of KD remains unknown and consequently there is no diagnostic test. The diagnosis is made using a constellation of clinical criteria that in isolation have poor sensitivity and specificity. Early treatment prevents overt coronary artery damage in the majority of children. The long-term effects of childhood KD on later cardiovascular health remain unknown. A recent study showed that treatment of KD in Australia is suboptimal, with late diagnosis occurring in approximately half of the cases and an unacceptably high incidence of acute cardiac involvement. These guidelines highlight the difficulties in the diagnosis of KD and offer some clues that may assist early recognition of this important paediatric disease. They also detail current treatment recommendations and the evidence on which they are based. Increased awareness of the epidemiology and spectrum of the clinical presentation of KD is essential for early recognition and optimal management. PMID- 15790317 TI - Airway gene therapy and cystic fibrosis. AB - Airway disease in cystic fibrosis (CF) is the major cause of death and is presently inadequately treatable, but genetic therapies offer the hope that such life-long disease will be curable, or at least satisfactorily treated. Normal pathogen defences that have evolved on airway surfaces also prevent the various gene vectors now available from producing effective gene transfer. Nevertheless, findings from basic research and human clinical trials are revealing how these barriers might be overcome or circumvented, with benefits to therapeutic efficacy and patient safety. Though progress is slower than expected or desired, the therapeutic rewards will be great when safe and effective gene therapy for CF airway disease becomes a clinical reality. PMID- 15790315 TI - Architecture of bacterial replication initiation complexes: orisomes from four unrelated bacteria. AB - Bacterial chromosome replication is mediated by single initiator protein, DnaA, that interacts specifically with multiple DnaA boxes located within the origin (oriC). We compared the architecture of the DnaA-origin complexes of evolutionarily distantly related eubacteria: two Gram-negative organisms, Escherichia coli and Helicobacter pylori, and two Gram-positive organisms, Mycobacterium tuberculosis and Streptomyces coelicolor. Their origins vary in size (from approx. 200 to 1000 bp) and number of DnaA boxes (from 5 to 19). The results indicate that: (i) different DnaA proteins exhibit various affinities toward single DnaA boxes, (ii) spatial arrangement of two DnaA boxes is crucial for the H. pylori and S. coelicolor DnaA proteins, but not for E. coli and M. tuberculosis proteins, and (iii) the oriC regions are optimally adjusted to their cognate DnaA proteins. The primary functions of multiple DnaA boxes are to determine the positioning and order of assembly of the DnaA molecules. Gradual transition from the sequence-specific binding of the DnaA protein to binding through co-operative protein-protein interactions seems to be a common conserved strategy to generate oligomeric initiator complexes bound to multiple sites within the chromosomal, plasmid and virial origins. PMID- 15790318 TI - Rheumatic fever: the relationship between clinical manifestations and laboratory tests. AB - OBJECTIVES: To determine the relationship between the severity of clinical features of rheumatic fever (RF), and antistreptolysin O titre (ASOT) and/or erythrocyte sedimentation rate (ESR). METHODS: Clinical and laboratory data from 102 children with RF who had been admitted at a university hospital in Tehran between 1992 and 2002 were reviewed retrospectively. In order to categorize the severity of clinical manifestations of disease, patients were divided into three groups. Those with arthritis alone were defined as group A, carditis with or without arthritis as group B and carditis (with or without arthritis) with congestive heart failure as group C. RESULTS: Thirty-one cases were enrolled in group A, 39 in group B, and 32 in group C. We didn't find a significant relationship between the severity of clinical presentation of disease and ASOT (P = 0.89) and ESR (P = 0.24). Seventy-two patients presented with first attack and 30 had recurrences. The frequency of congestive heart failure (CHF) in first attacks was 31.2%vs 68.8% in recurrences (P < 0.0001). No Significant relationship was found between the number of involved valves and ASOT (P = 0.4) or ESR (P = 0.8). CONCLUSION: Variable clinical presentation of disease and increasing intensity of cardiac involvement is not related to the ASOT or ESR levels. PMID- 15790319 TI - Impact of ethnicity upon body composition assessment in Sri Lankan Australian children. AB - OBJECTIVES: Obesity is a disease with excess body fat where health is adversely affected. Therefore it is prudent to make the diagnosis of obesity based on the measure of percentage body fat. Body composition of a group of Australian children of Sri Lankan origin were studied to evaluate the applicability of some bedside techniques in the measurement of percentage body fat. METHODS: Height (H) and weight (W) was measured and BMI (W/H(2)) calculated. Bioelectrical impedance analysis (BIA) was measured using tetra polar technique with an 800 microA current of 50 Hz frequency. Total body water was used as a reference method and was determined by deuterium dilution and fat free mass and hence fat mass (FM) derived using age and gender specific constants. Percentage FM was estimated using four predictive equations, which used BIA and anthropometric measurements. RESULTS: Twenty-seven boys and 15 girls were studied with mean ages being 9.1 years and 9.6 years, respectively. Girls had a significantly higher FM compared to boys. The mean percentage FM of boys (22.9 +/- 8.7%) was higher than the limit for obesity and for girls (29.0 +/- 6.0%) it was just below the cut-off. BMI was comparatively low. All but BIA equation in boys under estimated the percentage FM. The impedance index and weight showed a strong association with total body water (r(2)= 0.96, P < 0.001). Except for BIA in boys all other techniques under diagnosed obesity. CONCLUSIONS: Sri Lankan Australian children appear to have a high percentage of fat with a low BMI and some of the available indirect techniques are not helpful in the assessment of body composition. Therefore ethnic and/or population specific predictive equations have to be developed for the assessment of body composition, especially in a multicultural society using indirect methods such as BIA or anthropometry. PMID- 15790320 TI - Reduce the rads: a quality assurance project on reducing unnecessary chest X-rays in children with asthma. AB - OBJECTIVES: To quantify and then reduce the number of unnecessary chest X-rays (CXR) being performed on children presenting with asthma. METHODS: A retrospective review of case notes of all children, aged 1-15 years, who presented with asthma and had a CXR performed. The setting was two General Hospitals that see all children presenting to an emergency department in the region. The period of review was before and after the development and implementation of a simple guide for staff, with an education programme, outlining when CXR were deemed unnecessary (known asthmatic, primary diagnosis asthma, improving with treatment, pneumothorax not suspected, and not in Intensive Care Unit). RESULTS: In the 12 months prior to the education programme, 466 children presented with asthma: 260 had a CXR, of which 211 (81.1%) were unnecessary. During the 6 month period following implementation of the programme 197 presented with asthma: 72 had a CXR, of which 56 (78%) were deemed unnecessary. However the percentage of all children presenting with asthma who had an unnecessary CXR fell from 45.3% (211/466) to 28.4% (56/197): P = 0.00005. There was also a decrease in the admission rate from 46% before to 31% after the period of education. CONCLUSION: This study determined that an unacceptably high rate of unnecessary CXR was being ordered in children presenting to hospital with asthma. It also showed how a clinically and statistically significant reduction in the overall number of CXR could be achieved, through a simple and easy to implement educational programme. Further measures are needed in addition to ongoing education in order to improve on this achievement. PMID- 15790321 TI - Food-based recommendations to reduce fat intake: an evidence-based approach to the development of a family-focused child weight management programme. AB - OBJECTIVES: To develop food-based recommendations to lower fat and energy intake for use in a family-focussed weight management programme for 6-9 year old children. METHODS: Secondary analysis of the 1995 National Nutrition Survey (NNS95) informed the development of food-based recommendations aiming to reduce fat and energy intake. Each recommendation was used to progressively modify a model 3-day high fat dietary intake with the accumulative effect on energy and nutrient intake of each recommendation assessed. RESULTS: Six to nine-year-olds in the NNS95 consuming 35-45% energy as fat (n= 280) consumed more total energy (mean +/- SD, 8671 +/- 2741 vs. 7571 +/- 2328 kJ/day) than children consuming a 'low fat' (23-27% energy as fat, n= 85) diet (P < 0.002). Food-based recommendations found to be most effective for reducing energy and fat intake included; changing to reduced fat milk, reducing intake of cereal-based and snack foods and replacing juice or soft drink with water. These changes, together with avoiding adding fat to vegetables and using sources of lean meat, reduced energy intake by approximately 10%, total fat intake by approximately 30% and saturated fat intake by 53%. CONCLUSIONS: Modifying six areas of food choices results in a moderate reduction in fat and energy intake. An eating pattern that is consistent with Australian dietary guidelines and uses foods commonly eaten by children is achievable for children aged 6-9 years. These food-based recommendations provide an evidence-based dietary framework for prevention and management of overweight in children. PMID- 15790322 TI - Factors that influence special care nursery admissions to a district hospital in South-western Sydney. AB - OBJECTIVES: To explore the socio-demographic factors and maternal characteristics that influence special care nursery (SCN) admission for infants of more than 34 weeks' gestation. Particularly, this paper aims (i) to estimate the incidence of SCN admission by various mothers' socio-demographic factors; and (ii) to investigate the relationship between SCN admission and mothers' socio-demographic and obstetric and gynaecological factors. METHODS: This was a cohort study of 10,148 pregnant women who accessed the birthing unit within a public district hospital in south-western Sydney in New South Wales, between 1998 and 2001. The main outcome measure was risk factors for SCN admission. RESULTS: The incidence of SCN admission was 11.7%. Multivariate analysis revealed that the risk factors for SCN admission were diabetes, gestational diabetes, high parity, pregnancy induced hypertension, living in suburbs with low education and occupation index, and no private health insurance status. CONCLUSIONS: The results from this large population-based study suggest that, apart from clinical/medical factors, admission to a special care nursery at a District Hospital was significantly affected by maternal insurance status and level of education and occupation. PMID- 15790323 TI - Respiratory illness during winter: a cohort study of urban children from temperate Australia. AB - OBJECTIVE: To examine the epidemiology and burden of respiratory illness during winter in urban children from temperate Australia. METHODS: We conducted a cohort study of healthy Melbourne children, aged from 12 to 71 months. Parents kept a daily respiratory symptom diary and recorded resource use when an influenza-like illness (ILI) occurred. RESULTS: One-hundred and eighteen children had 137 ILI episodes over 12 weeks for a rate of 0.53 ILI episodes per child-month (95% CI 0.44-0.61). Risk factors for ILI included younger age, fewer people residing in the household, structured exposure to other children outside the home, and a higher household income. Episodes had a mean duration of 10.4 days with 64 visits to a general practitioner (46.7 GP visits per 100 episodes), 27 antibiotic courses prescribed (19.7 antibiotic courses per 100 episodes), and three overnight hospitalizations (2.2 admissions per 100 episodes). Parents reported an average of 11.7 h excess time spent caring for a child per episode. CONCLUSIONS: Respiratory illnesses are a common and largely neglected cause of illness in Australian children. Pathogen-specific data are required to better assess the likely impact of available and developing vaccines and other treatment options. PMID- 15790324 TI - Pre-ambulatory infants with head trauma: a three-year retrospective study. AB - OBJECTIVES: (i) To present the severity and documented mechanism of injury and clinical features of all infants less than 6 months old who presented with head trauma to a paediatric emergency department over a 3-year period; (ii) to evaluate the quality of their assessment; and (iii) to present the clinical features of those referred for assessment of possible inflicted injury. METHODS: Demographics, documented mechanism of injury, clinical features and management were retrospectively collected from the case notes. Adequacy of assessment was judged in terms of documentation of the history and examination, investigations performed and nature of any observation period. RESULTS: Two hundred and fifty one cases were analysed. Twenty-seven (11%) had sustained a major injury. Infants reported as dropped by a care giver were significantly more likely to have sustained a major injury. Infants with major injury were more likely to have vomited (P < 0.05), or to have external signs of injury (P < 0.0005). All patients with major injury had symptoms or signs or both. One hundred and thirty four infants had been inadequately assessed. Fourteen infants were referred for possible inflicted injury (confirmed in six cases). CONCLUSION: The mechanism of injury and clinical findings can provide some guide as to the likelihood of an infant having sustained a major injury after head trauma. The assessment of infants with head trauma could be improved if currently available guidelines for children under 2 years old were adopted. However, infant-specific guidelines should be developed. PMID- 15790325 TI - Haemophagocytic lymphohistiocytosis in Malaysian children. AB - OBJECTIVES: To study the clinical presentation, therapy and outcome of children diagnosed with both primary and secondary haemophagocytic lymphohistiocytosis (HLH) at the University of Malaya Medical Centre. METHODS: All patients diagnosed with HLH between 1998 and 2004 were studied. Clinico-pathological data of these patients were prospectively collected and analysed. RESULTS: Thirteen consecutive patients (eight boys) with a median age of 28 months were seen. All patients presented with high-grade unremitting fever and almost all, with hepatosplenomegaly and cytopenias. Neurological manifestations, which ranged from irritability to seizures and coma, were seen in 10 (77%) patients. Other common presenting features include liver dysfunction (46%) and skin rash (38%). All patients were treated using the HLH-94 protocol chemotherapy which consisted of a combination of etoposide, dexamethasone and cyclosporine. Complete response was seen in seven patients while two required bone marrow transplantation and one developed secondary acute myeloid leukaemia. Two patients died before treatment could be commenced. Overall mortality rate in our series was 46%. CONCLUSIONS: Haemophagocytic lymphohistiocytosis is an uncommon disease with a high fatality rate. Due to its protean clinical manifestations, it may be underdiagnosed. Early detection and prompt institution of appropriate therapy is necessary to improve the outcome in affected patients. PMID- 15790326 TI - Pertussis in the neonatal nursery. PMID- 15790327 TI - Anterior abdominal wall defects and biliary obstruction. AB - Three infants with anterior abdominal wall defects (gastroschisis and exomphalos) who presented with obstructive jaundice secondary to biliary obstruction, are described. All three infants had abnormal biliary systems, with mechanical distortion of the biliary tree. Biliary obstruction secondary to structural biliary anomalies should be considered in patients with abdominal wall defects and cholestasis, as prolonged unrelieved biliary obstruction may lead to biliary cirrhosis and portal hypertension. PMID- 15790328 TI - Cyclic bisphosphonate therapy in osteogenesis imperfecta type V. AB - The clinical and radiographic features and management of a young person with recently delineated Osteogenesis Imperfecta Type V is described. A female aged 9 years presented with a history of multiple fractures since 3 years of age and bilateral dislocation of the elbows from infancy. She was commenced on a low dose frequent regimen of cyclic intravenous pamidronate, which resulted in progressive improvement in bone density, reduced fracture frequency and remission of symptoms of osteoporosis. PMID- 15790329 TI - Neonatal cholinesterase inhibitor poisoning: materno-fetal exchange of pesticide. AB - This is a case report of a neonatal cholinesterase inhibitor poisoning after non accidental ingestion of a carbamate containing pesticide by the mother prior to delivery. The mother presented with symptoms of cholinesterase inhibitor poisoning at 40 weeks of gestation. A fetal bradycardia was present. The mother improved after administration of intravenous atropine. Spontaneous labour ensued and a term female infant weighing 3200 g was delivered. The infant had signs of cholinergic stimulation and decreased levels of serum cholinesterase. The neonate presented with a marked tachycardia. The heart rate normalized after administration of atropine. Therefore, we concluded that the tachycardia was due to the cholinesterase inhibitor substance, causing catecholamine release by stimulation of nicotinic receptors. Another potential contributing factor to the tachycardia, although unlikely in our patient, was the atropine administered to the mother prior to delivery. Mother and infant recovered well. We conclude that infants can present with tachycardia associated with cholinesterase inhibitor toxicity. Despite the side-effects intravenous atropine should not be withheld in the treatment of mother and infant. PMID- 15790330 TI - Hair-thread tourniquet syndrome. AB - We report a case of hair-thread tourniquet syndrome involving the labia majora of a 14-year-old autistic child. 'Hair-thread tourniquet' refers to the process whereby a thread of hair leads to the ischaemic strangulation of an appendage or other piece of tissue. Uncommonly recognized, this syndrome has gradually gained recognition since its first description in 1612. As it remains infrequent, many have attributed its aetiology to abuse or socio-cultural practices. We review the published literature on this infrequent but important paediatric condition. PMID- 15790331 TI - Acute meningococcaemia complicated by toxic epidermal necrolysis and acute supurative parotitis. PMID- 15790332 TI - Quality of life in childhood epilepsy: validating the QOLCE. PMID- 15790333 TI - Rapid rehydration. PMID- 15790340 TI - Epithelial cells in the female reproductive tract: a central role as sentinels of immune protection. AB - The continued presence of bacterial and viral antigens in the lumen of the vagina coupled with the periodic presence of antigens in the lumen of the upper reproductive tract provide an ongoing challenge that can compromise female reproductive health and threaten life. Separating underlying tissues from luminal antigens, polarized epithelial cells of the cervix, uterus and Fallopian tubes have evolved to protect against potential pathogens. Once thought to function exclusively by providing a crucial barrier, mucosal epithelial cells are now known to function as sentinels that recognize antigens, respond in ways that lead to bacterial and viral killing, as well as signal to underlying immune cells when pathogenic challenge exceeds their protective capacity. Unique to epithelial cells of the female reproductive tract is the regulatory control of the female sex hormones. Acting both directly and indirectly through underlying stromal cells, estradiol and progesterone regulate epithelial cell innate and adaptive immune functions to protect against potential pathogens while providing an environment that supports an allogeneic fetus. In this article, we will outline how polarized epithelial cells function as the first line of defense against potential pathogens in the female reproductive tract. PMID- 15790341 TI - Regulation and signal transduction of toll-like receptors in human chorioncarcinoma cell lines. AB - PROBLEM: Toll-like receptors (TLRs) are expressed on placental cells. The aim of this study is to analyze signaling components activated in placenta cells after TLR ligand engagement. METHODS OF STUDY: In chorioncarcimoma cell lines the regulation of TLRs was determined by real time polymerase chain reaction as well as by fluorescence-activated cell sorter analysis. Activation of NF-kappaB was determined in a reporter assay system and the activation of the mitogen-activated protein kinase signaling pathways by immunoblot analysis. RESULTS: Both lipopolysaccharide (LPS) and DNA oligonucleotides containing unmethylated CpG motifs (CpG) induced the enhanced expression of TLR2 mRNA as well as a TLR2 surface protein expression. Functionally, incubation of JAR cells with microbial stimuli such as LPS activated NF-kappaB, as well as the phosphorylation of ERK1/2 and p38 MAP kinases and secretion of interleukin-8. CONCLUSION: The functional expression of TLRs on placental cells may play an important role in the initiation of an immune response in the developing fetus. PMID- 15790342 TI - Endometriosis and systemic lupus erythematosus: a comparative evaluation of clinical manifestations and serological autoimmune phenomena. AB - PROBLEM: In view of evidences suggesting association between endometriosis (EM) and systemic lupus erythematosus (SLE), we have performed a comparative evaluation of clinical and humoral immunologic abnormalities in both diseases. METHOD OF STUDY: Forty-five women (18-40 years) with histologically confirmed pelvic EM, 21 healthy-women and 15 female SLE-patients (18-40 years) without surgically confirmed EM were prospectively evaluated. Immunologic investigations were performed by blinded researchers. RESULTS: None of the EM-patients fulfilled criteria for SLE. However, EM-patients presented higher frequencies of arthralgia (62%) and generalized myalgia (18%) superior than normal-controls (24%, P = 0.004/0%, P = 0.048) but comparable with SLE-patients (33%, P = 0.052/27%, P = 0.5). Similarly to SLE (7%), 9% of EM-patients presented fibromyalgia. Antinuclear antibodies (ANA) were detected in 18% of EM-sera, as compared with healthy-women (0%, P = 0.014) and SLE-patients (93%, P = 0.0005). In contrast with SLE, antibodies to dsDNA, Sm and U1RNP were negative in EM-sera. Anti-Ro and anticardiolipin antibodies were more often in SLE (40%, 33%) than in EM-patients (2%, P < 0.001/9%, P = 0.04). Elevated immune-complexes and low total complement were more frequent in SLE (40%, 13%) compared with EM-sera (7%, P = 0.005/0%, P = 0.01). CONCLUSIONS: Our data indicate differences of ANA antigenic specificity and complement consumption between EM and SLE. The high prevalence of generalized musculoskeletal complaints in EM justifies a multidisciplinary approach. PMID- 15790343 TI - A case-control study in IL6 and TGFB1 gene polymorphisms and recurrent spontaneous abortion in southern Brazilian patients. AB - PROBLEM: A high proportion of recurrent spontaneous abortions (RSA) remains unexplained. Cytokine genotyping has been investigated. We studied the relationship between unexplained RSA, IL6 (-174 G-->C) and TGFB1 (+869, T-->C; +915, G-->C) gene polymorphisms. METHOD OF STUDY: The case-control study composed of 57 south Brazilian women, with unexplained RSA and 74 controls carefully matched was performed. Cytokine genotyping was performed by the polymerase chain reaction-sequence specific primer method, using the 'Cytokine Genotyping Tray'. RESULTS: The results showed that the genotypic frequencies did not differ from samples for TGFB1 gene. In relation to IL6 gene polymorphism there was a statistical difference in genotypic distribution between samples (P < or = 0.025). The frequency of the C/C genotype was increased in women with RSA in comparison with the frequency observed in controls: 18% versus 4% (P = 0.01). CONCLUSIONS: This result strengthened the importance of IL6 genotypes in the pathogenesis of RSA of unknown cause in the south Brazilian population. PMID- 15790344 TI - Association of the A/G polymorphism at position 49 in exon 1 of CTLA-4 with the susceptibility to unexplained recurrent spontaneous abortion in the Chinese population. AB - PROBLEM: To investigate whether the A/G polymorphism at position 49 in exon 1 of cytotoxic T lymphocyte antigen-4 (CTLA-4) gene, which delivers a negative signal to T-cell activation, confers the susceptibility to unexplained recurrent spontaneous abortion in the Chinese population. METHOD OF STUDY: A total of 168 patients with unexplained recurrent spontaneous abortion (RSA), who were treated in the Renji Hospital affiliated to the Shanghai Second Medical University, were matched against 117 women with normal pregnancy history. Case-control study to compare the frequency of G/A alleles, AA/AG/GG genotypes and A + (AA + AG) /G+ (GG + AG) phenotypes of CTLA-4 between RSA patients and controls were performed. After amplification of CTLA-4 exon-1 region by polymerase chain reaction (PCR), restriction fragment-length polymorphism (RFLP) was used to detect the polymorphism at position 49 in exon-1 of CTLA-4 gene. Statistical significance was tested by SPSS software. RESULTS: There were dissimilar distributions of G/A alleles, AA/AG/GG genotypes and A+/G+ phenotypes of CTLA-4 between RSA patients and controls. The frequencies of G allele (P = 0.032) and GG genotype (P = 0.011) in RSA patients were significantly higher than those in controls, while the frequencies of AG genotype (P = 0.039) and A + (AA + AG) phenotype in RSA patients were decreased significantly (P = 0.011). CONCLUSIONS: Our findings suggest that A/G polymorphism in exon-1 of CTLA-4 is associated with the immunopathogenesis of RSA, and it confers susceptibility to RSA in Chinese population. PMID- 15790346 TI - Autoimmunity. PMID- 15790345 TI - Antibodies against granulosa luteinized cells and their targets in women attending IVF program. AB - PROBLEM: The study was conducted to investigate the presence and the possible role of anti-granulosa luteinized cells (GLCs) antibodies in infertile patients, attending in vitro fertilization (IVF) cycles, as well as to attempt the identification of putative target antigens. METHOD OF STUDY: A total of 101 sera from infertile women (patient group) and 33 sera from fertile women (control group) were investigated. The patients were divided in subgroups according to: ovarian response to controlled ovarian hyperstimulation (COHS)--poor responders and good responders; fertilization rate--low fertilization rate and high fertilization rate. The relation between embryo transfer (ET) outcome and prevalence of anti-GLC antibodies was investigated too. NONIDET P-40 (NP-40) and whole-GLC lysates, as antigenic materials, were used in enzyme-linked immunosorbent assay (ELISA) and immunoblotting, respectively. RESULTS: Based on the cut-off value of 2 S.D. above the control mean value, the prevalence of patients positive for anti-GLC antibodies was found to be 28.7% (29/101) compared with 9.1% (3/33) in control fertile women (P < 0.05). Anti-GLC antibodies were detected in 54.8% (17/31) of women with low fertilization rate compared with 15.7% (11/70) of women with high fertilization rate (P = 0.003) and it was established that women positive for anti-GLC antibodies had lower fertilization rate, than those negative for anti-GLC antibodies. Immunoblotting has demonstrated a number of GLC proteins with molecular masses (MM) of 110, 70-80, 47 and 37 kDa, frequently reacting with anti-GLC antibodies in patient's sera, as possible targets. CONCLUSION: The presented results on the prevalence and role of anti-GLC antibodies, as well as the data on the target antigens confirmed our hypothesis about the potential involvement of GLC antigens in some cases of immunological infertility in IVF patients. PMID- 15790347 TI - Interferons as pathogenic effectors in autoimmunity. AB - Interferons (IFNs) type-1 (IFN alpha/beta) and type-II (IFN-gamma) are the most pleiotropic molecules in the intricate cytokine network. This dominance arises from three crucial factors: (i) initiation of IFN-alpha/beta and IFN-gamma production at the inception of most innate immune responses, which primes for the ensuing adaptive immune responses, primarily through the sine qua non upregulation of major histocompatibility complex and costimulatory molecules; (ii) magnification of their production and signaling by cross-talk between themselves, and synergistic or antagonistic effects on other cytokines; and (iii) direct or indirect initiation of transcription of hundreds of immunologically relevant genes. Considering that aberrant immune responses against self-molecules seem to depend on the same constituents and pathways as those against exogenous antigens, it follows that IFNs are also major effectors in the pathogenesis of autoimmunity. Here, we review the diverse biological effects of IFNs on the immune system, discuss findings pertaining to the nature of exogenous and endogenous stimuli that might induce IFN production through the engagement of Toll-like receptors, and summarize the detrimental and, in some instances, beneficial effects of IFNs in systemic and organ-specific autoimmune diseases. PMID- 15790348 TI - Toll-like receptors, endogenous ligands, and systemic autoimmune disease. AB - The critical role of Toll-like receptors (TLRs) as mediators of pathogen recognition by the innate immune system is now firmly established. Such recognition results in the initiation of an inflammatory immune response and subsequent instruction of the adaptive immune system, both of which are designed to rid the host of the invading pathogen. More controversial is the potential role of TLRs in the recognition of endogenous ligands and what effect this might have on the consequent development of autoimmune or other chronic sterile inflammatory disorders. An increasing number of studies implicate TLRs as being involved in the immune response to self-molecules that have in some way been altered from their native state or accumulate in non-physiologic sites or amounts, although questions have been raised about possible contaminants in certain of these studies. In this review, we discuss the evidence for endogenous ligand-TLR interactions with particular emphasis on mammalian chromatin, systemic lupus erythematosus, and atherosclerosis. Overall, the data support the general concept of a role for TLRs in the recognition of endogenous ligands. However, the precise details of the interactions and the extent to which they may contribute to the pathogenesis of human disease remain to be clarified. PMID- 15790349 TI - The role of BAFF in immune function and implications for autoimmunity. AB - BAFF [B-cell activating factor belonging to the tumor necrosis factor (TNF) family] is a ligand that is required for peripheral B-cell survival and homeostasis. In addition to mediating B-cell survival, BAFF also regulates expression of certain B-cell-surface proteins, such as CD21/35. BAFF deficiency results in a reduced number of peripheral B cells and a diminished ability to mount robust humoral immune responses. Overexpression of BAFF has been linked to murine and human autoimmunity, and recent data provide clues as to how excess BAFF may allow the emergence of autoreactive B cells. In vivo animal testing with BAFF inhibitors has generated exciting data that support the pathway as a target for modulating B cells. The role of BAFF in B-cell biology, T-cell biology, and autoimmunity is discussed, as well as current efforts to develop BAFF inhibitors for clinical testing in autoimmune disorders. PMID- 15790350 TI - Rheumatoid arthritis. AB - Therapeutic efficacy of depleting B cells or blocking T-cell costimulation in rheumatoid arthritis (RA) has confirmed the critical pathogenic role of adaptive immune responses. Yet, RA preferentially affects elderly individuals, in whom adaptive immunity to exogenous antigens begins to fail. Here, we propose that senescence of the immune system is a risk factor for RA, with chronic inflammation resulting from the accumulation of degenerate T cells that have a low threshold for activation and utilize a spectrum of novel receptors to respond to microenvironmental cues. The process of immunosenescence is accelerated in RA and precedes the onset of disease, the acceleration, in part, being conferred by the HLA-DR4 haplotype. Naive CD4(+) T cells in RA are contracted in diversity and restricted in clonal burst. Senescence of effector CD4(+) T cells is associated with the loss of CD28 and the de novo expression of KIR2DS2, NKG2D, and CX(3)CR1, all of which function as costimulatory molecules and reduce the threshold for T cell activation. The synovial microenvironment promotes chronic persistent immune responses by facilitating ectopic lymphoid neogenesis, such as the formation of aberrant germinal centers. With the propensity to develop complex lymphoid architectures and to provide optimal activation conditions for senescent CD4(+) T cells, the synovium becomes a natural target for pathogenic immune responses in prematurely aged individuals. PMID- 15790351 TI - Pathways to gene identification in rheumatoid arthritis: PTPN22 and beyond. AB - Rheumatoid arthritis (RA), like other autoimmune diseases, has a complex genetic basis. Rapid technical advances in high-throughput genotyping and analysis have now reached a point where genes of low-to-moderate risk can be identified using a variety of study designs, including whole genome association studies. The availability of large, well-characterized populations of cases and controls are critical to the success of these efforts. A functional variant (R620W) of the intracellular protein tyrosine phosphatase N22 (PTPN22) has now been conclusively shown to confer approximately two-fold risk for seropositive RA as well as several other autoimmune disorders. PTPN22 appears to act primarily by setting thresholds for T-cell receptor signaling, and the current data suggest that the PTPN22 620W allele is likely to be a general risk factor for the development of humoral autoimmunity. PTPN22 is expressed widely in hematopoietic cells, but other than in T cells, its role is unknown. These results provide strong evidence for the longstanding hypothesis that common genes underlie different autoimmune phenotypes and emphasize that finding genes of only moderate risk can provide important insights into disease pathogenesis. PMID- 15790352 TI - Genetic lesions in T-cell tolerance and thresholds for autoimmunity. AB - The cause of common organ-specific autoimmune diseases is poorly understood because of genetic and cellular complexity in humans and animals. Recent advances in the understanding of the mechanisms of the defects underlying autoimmune disease in autoimmune polyendocrinopathy syndrome type 1 and non-obese diabetic mice suggest that failures in central tolerance play a key role in predisposition towards organ-specific autoimmunity. The lessons from such rare monogenic autoimmune disorders and well-characterized polygenic traits demonstrate how subtle quantitative trait loci can result in large changes in the susceptibility to autoimmunity. These data allow us to propose a model relating efficiency of thymic deletion to T-cell tolerance and susceptibility to autoimmunity. PMID- 15790353 TI - CTLA4 gene polymorphism and autoimmunity. AB - CD28 and cytotoxic T-lymphocyte antigen-4 (CTLA4) are two receptors that have critical but opposing functions in T-cell stimulation. CD28 promotes a number of T-cell activities, whereas in contrast CTLA4 is an essential inhibitor of T-cell responses. Because of its inhibitory role, CTLA4 is a strong candidate susceptibility gene in autoimmunity and several studies suggest disease associated polymorphisms. In this review, we discuss recent progress in relating CTLA4 polymorphisms to disease susceptibility and consider the putative mechanisms by which CTLA4 may act to inhibit autoimmunity. PMID- 15790354 TI - T-cell tolerance and autoimmunity to systemic and tissue-restricted self antigens. AB - We have used transgenic mouse models to examine the mechanisms of tolerance in CD4(+) T lymphocytes to soluble, systemic and cell-associated, tissue-restricted self-antigens. Anergy to an islet antigen, as a model of a tissue antigen, is dependent on the inhibitory receptor cytotoxic T-lymphocyte antigen-4 (CTLA-4), and tissue-restricted autoimmunity is inhibited by regulatory T lymphocytes. Anergy to a circulating systemic antigen can occur independently of CTLA-4 signals, and it is induced primarily by a block in proximal receptor-initiated signals. CD4(+)CD25(+) regulatory T cells are generated in response to both forms of self-antigens, but the induction is much more efficient with the tissue antigen. Receptor desensitization can be induced by the systemic antigen even in the absence of regulatory T cells, but tolerance can be broken by immunization much more easily if these cells are absent. Deletion of mature T cells is striking with the systemic antigen; there is little evidence to support peripheral deletion as a mechanism of tolerance to the tissue antigen. Thus, both distinct and overlapping mechanisms account for unresponsiveness to different forms of self-antigens. These results establish a foundation for searching for genetic influences and pathogenic mechanisms in organ-specific and systemic autoimmune diseases. PMID- 15790355 TI - The B7/CD28 costimulatory family in autoimmunity. AB - Host defense is dependent on the appropriate induction of immune responses. A central concept in immunology is the ability of the immune system to differentiate foreign from self-antigens. The failure of the immune response to recognize foreign pathogens can result in infection and disease in the host. The inappropriate response of the immune system to self-antigens is equally problematic, leading to autoimmune disease. Central and peripheral tolerance mechanisms control self-reactive T-cell responses and protect peripheral tissues from autoimmune attack. This review examines the roles of B7/CD28 family members, which can augment or antagonize T-cell receptor signaling, in the regulation of central and peripheral T-cell tolerance. We also discuss how B7/CD28 pathways influence both T-cell-intrinsic and -extrinsic mechanisms of regulation. PMID- 15790356 TI - Tumor necrosis factor family members and inflammatory bowel disease. AB - Tumor necrosis factor (TNF) is one of the most potent effector cytokines in the pathogenesis of inflammatory bowel disease (IBD). Previous studies strongly implicate the critical involvement of several TNF family members in human IBD. This review focuses on the recent studies of TNF family members in IBD development. In particular, we discuss the findings about LIGHT (homologous to lymphotoxins, inducible expression, competes with herpes simplex virus glycoprotein D for herpes viral entry mediator, a receptor expressed on T lymphocytes) in the pathogenesis of IBD, and the potential mechanisms by which LIGHT induces IBD. Such mechanisms may also apply to other TNF family members. PMID- 15790357 TI - AIRE and APECED: molecular insights into an autoimmune disease. AB - Mutations in the autoimmune regulator (AIRE) protein are the causative factor in development of the human disease autoimmune polyendocrinopathy-candidiasis ectodermal dystrophy (APECED). In mice, the absence of the analogous protein aire influences ectopic expression of peripheral tissue antigens in thymic medullary epithelial cells (MECs), resulting in the development of an autoimmune disorder similar to APECED and establishing aire/AIRE as an important player in the induction of central tolerance. However, the molecular mechanism of AIRE's function, in particular its ability to specifically control the expression of peripheral tissue antigens in MECs, is still unclear. Here, we review current evidence relating to the molecular mechanism of AIRE. PMID- 15790358 TI - The regulation and activation of lupus-associated B cells. AB - Anti-double-stranded DNA (anti-dsDNA) B cells are regulated in non-autoimmune mice. While some are deleted or undergo receptor editing, a population of anti dsDNA (VH3H9/V lambda 1) B cells that emigrate into the periphery has also been identified. These cells have an altered phenotype relative to normal B cells in that they have a reduced lifespan, appear developmentally arrested, and localize primarily to the T/B-cell interface in the spleen. This phenotype may be the consequence of immature B cells encountering antigen in the absence of T-cell help. When provided with T-cell help, the anti-dsDNA B cells differentiate into antibody-forming cells. In the context of the autoimmune-prone lpr/lpr or gld/gld mutations, the VH3H9/V lambda 1 anti-dsDNA B cells populate the B-cell follicle and by 12 weeks of age produce serum autoantibodies. The early event of anti dsDNA B-cell follicular entry, in the absence of autoantibody production, is dependent upon CD4(+) T cells. We hypothesize that control of autoantibody production in young autoimmune-prone mice may be regulated by the counterbalancing effect of T-regulatory (T(reg)) cells. Consistent with this model, we have demonstrated that T(reg) cells are able to prevent autoantibody production induced by T-cell help. Additional studies are aimed at investigating the mechanisms of this suppression as well as probing the impact of distinct forms of T-cell-dependent and -independent activation on anti-dsDNA B cells. PMID- 15790359 TI - Regulatory T cells and intestinal homeostasis. AB - Murine models of inflammatory bowel disease (IBD) are useful tools for the study of the pathogenesis and regulation of intestinal inflammation. Colitis can be induced in immune-deficient mice following transfer of populations of T cells or following infection with Helicobacter hepaticus and other intestinal pathogens. In these situations, colitis occurs as a result of the absence of a specialized population of regulatory cells, as transfer of CD4(+)CD25(+) T cells prevents disease. Importantly, from a clinical perspective, CD4(+)CD25(+) T cells can also reverse an established colitis. CD4(+)CD25(+) T cells proliferate both in the secondary lymphoid organs and at the site of inflammation, suggesting that regulation occurs both locally and systemically. CD4(+)CD25(+) T cells are not only capable of regulating other T cells but are also capable of suppressing components of the innate immune system. Control of colitis is dependent on the presence of the immunosuppressive cytokines interleukin-10 and transforming growth factor-beta, although their roles are divergent and complex. Regulatory T cells represent one of the host's mechanisms to prevent immune pathology during chronic immune stimulation. Enhancement of regulatory T-cell activity may be useful to control autoreactive T-cell responses and inhibit harmful inflammatory diseases such as asthma and IBD. PMID- 15790360 TI - Regulatory T cells and autoimmune disease. AB - Although T-cell clones bearing T-cell receptors with high affinity for self peptide major histocompatibility complex (MHC) products are generally eliminated in the thymus (recessive tolerance), the peripheral T-cell repertoire remains strongly biased toward self-peptide MHC complexes and includes autoreactive T cells. A search for peripheral T cells that might exert dominant inhibitory effects on autoreactivity has implicated a subpopulation of CD4(+)CD25(+) T cells called regulatory T cells (Tregs). Here, we discuss the role of cytokines and costimulatory molecules in the generation, maintenance, and function of Tregs. We also summarize evidence for the involvement of Tregs in controlling autoimmune diseases, including type 1 diabetes, experimental autoimmune encephalomyelitis, and inflammatory bowel disease. Last, we discuss our recent definition of the potential role of B7 expressed on activated T-effector cells as a target molecule for Treg-dependent suppression. These observations suggest that the engagement of B7 on effector T cells transmits an inhibitory signal that blocks or attenuates effector T-cell function. We restrict our comments to the suppression mediated by cells within the CD4 lineage; the impact of the cells within the CD8 lineage that may suppress via engagement of Qa-1 on effector T cells is not addressed in this review. PMID- 15790361 TI - Multiple sclerosis. AB - Multiple sclerosis (MS) is a complex genetic disease associated with inflammation in the central nervous system (CNS) white matter and is thought to be mediated by autoimmune processes. Clonal expansion of B cells, their antibody products, and T cells, hallmarks of inflammation in the CNS, are found in MS. The association of the disease with major histocompatibility complex genes, the inflammatory white matter infiltrates, similarities with animal models, and the observation that MS can be treated with immunomodulatory and immunosuppressive therapies support the hypothesis that autoimmunity plays a major role in the disease pathology. This review discusses the immunopathology of MS with particular focus given to regulatory T cells and the role of B cells and antibodies, immunomodulatory therapeutics, and finally new directions in MS research, particularly new methods to define the molecular pathology of human disease with high-throughput examination of germline DNA haplotypes, RNA expression, and protein structures that will allow the generation of a new series of hypotheses that can be tested to develop better understandings and therapies for this disease. PMID- 15790363 TI - The good turned ugly: immunopathogenic basis for diabetogenic CD8+ T cells in NOD mice. AB - Type 1 diabetes (T1D) in both humans and nonobese diabetic (NOD) mice is a T-cell mediated autoimmune disease in which the insulin-producing pancreatic islet beta cells are selectively eliminated. As a result, glucose metabolism cannot be regulated unless exogenous insulin is administered. Both the CD4(+) and the CD8(+) T-cell subsets are required for T1D development. Approximately 20 years ago, an association between certain class II major histocompatibility complex (MHC) alleles and susceptibility to T1D was reported. This finding led to enormous interest in the CD4(+) T cells participating in the development of T1D, while the CD8(+) subset was relatively ignored. However, the isolation of beta cell-autoreactive CD8(+) T-cell clones from the islets of NOD mice helped to generate interest in the pathogenic role of this subset, as has accumulating evidence that certain class I MHC alleles are additional risk factors for T1D development in humans. Three distinct diabetogenic CD8(+) T-cell populations have now been characterized in NOD mice. Here, we review recent investigations exploring their selection, activation, trafficking, and antigenic specificities. As CD8(+) T cells are suspected contributors to beta-cell demise in humans, continued exploration of these critical areas could very possibly lead to tangible benefits for T1D patients and at-risk individuals. PMID- 15790362 TI - The stages of type 1A diabetes: 2005. AB - Type 1A diabetes is a chronic autoimmune disease usually preceded by a long prodrome during which autoantibodies to islet autoantigens are present. These antibodies are directed to a variety of antigens, but the best characterized are glutamic acid decarboxylase-65, insulinoma-associated antigen-2, and insulin. We hypothesize that the natural history of type 1A diabetes can be represented by several stages, starting from genetic susceptibility and ending in complete beta cell destruction and overt diabetes. Type 1A diabetes probably results from a balance between genetic susceptibility and environmental influences. In both humans and animal models, the major determinants of the disease are genes within the major histocompatibility complex. The next best-characterized susceptibility locus is the insulin gene, the variable nucleotide tandem repeat locus. This gene affects the expression of insulin in the thymus and thus may play a role in the modulation of tolerance to this molecule. In a subset of genetically susceptible individuals, the activation of autoimmunity may be triggered by environmental factors such as viruses and/or diet. However, no conclusive association has been established between type 1A diabetes and specific environmental triggers. In this review, we provide evidence that insulin has a fundamental role in anti-islet autoimmunity. PMID- 15790364 TI - Protein arrays for studying blood cells and their secreted products. AB - Protein microarrays have been developed and partially validated for studying blood cells, which play a role in many human diseases. Arrays of capture antibodies are commercially available for analyzing cytokines and intracellular signaling proteins. Several academic laboratories have developed antigen microarrays for characterizing autoimmune and allergic diseases, with a goal toward using such arrays to profile antibodies found in blood or other biological fluids. Arrays composed of major histocompatibility complex tetramers have been constructed and validated for analysis of immune responses in mice, paving the way toward studying antigen-specific T-lymphocyte responses. Finally, reverse phase protein lysate microarray technology, first developed for analyzing cancer cells from tissue sections, has now been demonstrated for studying living cells, including knockout cells, cells treated with drugs such as kinase inhibitors, and rare populations of lymphocytes such as regulatory T cells. The goal of this review is to focus on advances in and future uses of arrays of proteins that can be printed on glass microscope slides using traditional microarray robots that are commonly found at academic medical centers. Dissemination of protein array technology will occur in the next decade and will markedly change how immunology research, particularly in the fields of autoimmunity and inflammation, is conducted. PMID- 15790365 TI - Prevalence of oral mucosal lesions in children and youths in the USA. AB - INTRODUCTION: There is a dearth of studies of oral lesions in children and youths using probability samples of a general population. The present paper describes the results of the Third National Health and Nutrition Examination Survey, 1988 1994 (NHANES III), and compares them to those of the National Survey of Oral Health in US Schoolchildren, 1986-1987. METHODS: The NHANES III was a large US study based on a multistage probability sample. Dentist examiners were trained to recognize, classify and record, in a standard manner, the clinical characteristics of each of the 48 conditions of interest using procedures based on the World Health Organization's Guide to Epidemiology and Diagnosis of Oral Mucosal Diseases. RESULTS: Examinations were performed on 10,030 individuals (10.26%) aged between 2 and 17 years, 914 of whom had a total of 976 lesions. The lip was the most frequent site of lesions (30.7%), followed by the dorsum of the tongue (14.7%) and the buccal mucosa (13.6%). Lesions were more prevalent in males (11.76%) than females (8.67%). The most prevalent lesions were lip/cheek bite (1.89%), followed by aphthous stomatitis (1.64%), recurrent herpes labialis (1.42%) and geographic tongue (1.05%). The prevalence of recurrent aphthous stomatitis in the NHANES III child and youth survey was substantially higher than that for adults, while the NHANES III adult estimates for geographic tongue (1.85%; 95% CI 1.42, 2.28) and cheek/lip bite (3.05%; 95% CI 2.36, 3.74) were substantially greater than those for children and youths (0.97% and 2.05%, respectively). CONCLUSION: Reported prevalences for rare conditions in other studies employing more selected samples (especially if standard errors or confidence intervals are not provided) should be interpreted with caution. Studies of adult populations, however valid, may have limited applicability to children. PMID- 15790366 TI - Dental arch morphological and dimensional characteristics in Jordanian children and young adults with beta-thalassaemia major. AB - OBJECTIVE: The aim of this study was to examine the arch dimensions of Jordanian patients with beta-thalassaemia major in comparison with an unaffected control group. METHODS: The sample consisted of 24 patients who suffered from beta thalassaemia major (mean age = 13.9 +/- 3.1 years) and an unaffected control group (mean age = 13.5 +/- 2.9 years) matched for dental age, sex, and incisor and molar relationships. The unaffected control group was randomly selected from four public schools in the Governate of Irbid-Jordan. Alginate impressions were taken of the maxillary and mandibular dental arches of all participants. All measurements of the arch dimensions were made on the casts using an electronic digital sliding calliper. RESULTS: In the mandibular arch, when compared with the patients with thalassaemia, the unaffected control group subjects showed a (statistically) significantly larger incisor width, larger arch depth, and larger left and right anterior arch lengths (1.18, 2.58, and 1.85 and 1.12 mm, respectively). In the maxillary arch, there was a statistically significant difference in the mean incisor width (+/- 2.16 mm), arch depth (+/- 3.14 mm), inter-molar width (+/- 1.21 mm) and in the left anterior arch length (+/- 1.97 mm). The canine widths, premolar widths, left and right posterior arch length, and curve of Spee of both arches showed no statistically significant differences between the two groups. CONCLUSION: When compared to unaffected subjects, patients with beta-thalassaemia major exhibited: a narrower maxilla; a shorter maxilla and mandible; and smaller incisor widths for the maxillary and mandibular arches. PMID- 15790367 TI - Prevalence and impact of oral pain in 8-year-old children in Sri Lanka. AB - OBJECTIVE: The present study aimed to determine the prevalence and impact of oral pain in 8-year-old Sri Lankan children and their parents. DESIGN: A cross sectional study using a multistage cluster sampling technique was carried out in a sample of 614 children attending schools in the Education Division of Badulla, Uva province, Sri Lanka. Data were collected by means of a pre-tested questionnaire that was sent to each of the children and their parents. The children were also given an oral examination. Of the 614 questionnaires which were sent to parents, 30 were not returned and eight had to be excluded from the analysis because the respondents had answered only two questions. Therefore, the present analysis is limited to data from 576 children and their parents. RESULTS: The lifetime prevalences of oral pain, as reported by the children and parents, were 49% and 53%, respectively. According to the children's report, 25% had experienced oral pain in the past 2 months while 31% of the parents reported that their child had experienced oral pain within the same period. Of those children who reported that they had experienced oral pain in the past 2 months, nearly 45% stated that the pain was severe. In 48%, the pain was triggered when biting. 'Presence of cavity/decayed tooth' was cited as the most common cause of oral pain (67%). Overall, 74% of children had experienced a negative impact as a result of the pain whilst 66% of the parents said that the child's pain had had an impact on them. Ethnic group, parental income and level of education, and the caries experience of the child were significantly associated with the reporting of oral pain in the past 2 months. CONCLUSIONS: The prevalence of oral pain was high among these children, and had a considerable impact on both the children and their parents. PMID- 15790369 TI - Oral lichen planus in childhood: a report of three cases. AB - Lichen planus is a common mucocutaneous condition in which the occurrence of oral lesions has been cited as between 0.5 and 1.9% of the adult population. It is rare in childhood. Here we report three cases of children with intra oral lesions of lichen planus. Lichen planus, although reportedly rare in childhood, should be considered in the diagnosis of hyperkeratotic or erosive lesions of the oral mucosa in children. PMID- 15790368 TI - Takayasu's arteritis: what should the dentist know? AB - Takayasu's arteritis is a chronic inflammatory disease that affects large blood vessels, especially the aorta and/or its major branches. The condition presents with segmental lesions adjacent to normal, apparently unaffected, areas. The lesions include stenosis, occlusion, dilatations or aneurysm formations along the path of the affected artery. Because of the severity of the disease and the possibility of cardiovascular complications, patients with Takayasu's arteritis require medical treatment based on immunosuppressive and antihypertensive drugs, as well as regular follow up and surgical intervention in many instances. The aim of this paper was to describe the characteristics of Takayasu's arteritis, to report dental treatment carried out on an affected patient, and to discuss the main implications and care required during routine treatment for children in the dental office. PMID- 15790370 TI - Tooth development included in the multifocal jaw lesions of Langerhans cell histiocytosis. AB - Many cases demonstrating the oral manifestations of Langerhans cell histiocytosis (LCH) have been reported; however, tooth development in jaw lesions has rarely been mentioned. This paper reports the case of a 3-year-old Japanese girl with LCH suffering from multiple osteolytic lesions of the skull and jaw bones. She was referred to our paediatric clinic because of swelling of the mucogingival folds in the upper and lower primary molar regions. The patient responded well to steroid therapy and the osteolytic lesions resolved. There was an accompanying development of permanent tooth germs included in the lesions. Langerhans cell histiocytosis in children usually has a long-term clinical course and paediatric dentists should be involved with oral health care for affected patients. PMID- 15790371 TI - The dental management of a patient with hyperimmunoglobulinemia E syndrome: a case report. AB - Hyperimmunoglobulinemia E recurrent infection syndrome (also known as Job's syndrome) is a rare multi-system primary immunological disorder in which non immunological abnormalities of the dentition, bones and connective tissue are also seen. A previous study has reported the occurrence of dental abnormalities in three-quarters of individuals diagnosed as suffering from this condition. The present authors report the case of a boy whose prolonged retention of the primary dentition was associated with delayed eruption of permanent teeth. They emphasize the need for early intervention in order to help minimize later orthodontic problems. PMID- 15790372 TI - Sturge-Weber syndrome in a 6-year-old girl. AB - Sturge-Weber syndrome is a congenital disorder characterized by vascular facial birthmarks and neurological abnormalities. Oral cavity involvement may occur, and the extent of the vascular abnormality may vary considerably. The present authors report the case of a 6-year-old girl with Sturge-Weber syndrome, focusing on the clinical and radiographic features. Her dental management involved a multidisciplinary team and included orthodontic treatment using removable appliances. PMID- 15790373 TI - Eczema herpeticum: a case report. AB - Primary herpetic gingivostomatitis is a relatively common and well-recognized condition that the dental practitioner may encounter in clinical practice. A primary herpetic infection in a child with atopic dermatitis such as eczema, however, predisposes them to eczema herpeticum. This can be a severe and potentially life-threatening condition. This paper describes one such case, discusses the aetiology, presentation and management of this condition, and highlights the importance of early recognition by the clinician. PMID- 15790374 TI - Noonan syndrome with giant cell lesions. AB - Noonan syndrome is characterised by short stature, unusual facies, congenital heart disease, chest deformity and mild mental retardation. It may be sporadic or inherited as an autosomal dominant trait and occurs between one in 1000-2500. Cherubism is a giant cell lesion of the jaws thought to be transmitted as an autosomal dominant trait. It is usually recognised by age two to four years, follows a variable course, and is not known to be related to other genetic disorders. The purpose of this article is to report a case of multiple giant cell lesions of the mandible that occurred in a patient with phenotypic features of Noonan syndrome. The emerging relationship between these cherubism-like findings and Noonan syndrome will be discussed. PMID- 15790375 TI - Junctional epidermolysis bullosa: a case report. AB - Epidermolysis bullosa (EB) is a group of genetically determined disorders characterized by blistering of the skin and mucosae. There are three major forms- simple, junctional and dystrophic--and each has several varieties. The present case report describes a male child with junctional EB. The aim of the report is to present the dietary situation and the dental status of the child, examples of potential dental and nutritional consequences, and the therapeutic interventions possible for children with this disease. PMID- 15790376 TI - The influence of orthognathic surgery on masticatory performance in retrognathic patients. AB - Surgical correction of retrognathism may influence chewing performance and its underlying mechanisms selection and breakage of food particles. In this study we examined the hypothesis that treatment of this anomaly improves chewing performance. Furthermore, we determined to what extent this change can be attributed to selection and breakage of food particles. Eleven patients were tested before and 1-1.5 years after surgery. To determine chewing performance, eight cubes of 8 mm of a silicone rubber (Optosil) were used as a test food. Selection and breakage were determined in one-chew experiments using three particle sizes. On average, no differences were found for chewing performance, selection or breakage. However, on an individual basis, patients with a poor chewing performance before surgery tended to improve, whereas no improvement was observed for patients with a good chewing performance. The change in chewing performance was mainly due to a change in breakage of the food particles. PMID- 15790377 TI - Is there a greater mandibular movement capacity towards the left? Verification of an observation from 1921. AB - In 1921, the German dentist Hans Wertheim reported that more individuals were able to shift the mandible more towards the left than to the right. This study analyses the deviation from symmetrical mobility of the lower jaw in either direction. Using a millimetre ruler, maximum jaw opening (MJO), maximum left laterotrusion (MLL), and maximum right laterotrusion (MRL) were recorded in 141 healthy individuals and in 141 patients with temporomandibular disorders (TMDs). For both sexes, the mean maximum movements to the left and to the right were greater in the healthy group as compared with the TMD group. Healthy subjects as well as patients were able to move the mandible more to one side. Only a minority had identical values for MLL and MRL. The majority of healthy individuals and TMD patients could move more to the left (P < 0.001). In the healthy group, the mean ratio between MJO and MLL was 5.0, and 5.5 between MJO and MRL. In the TMD group, the corresponding values were 4.6 and 6.1. The mean absolute difference between MLL and MLR (in mm) was 1.24 [95% confidence interval (CI): 0.99; 1.49] among healthy females, and 2.09 (95% CI: 1.52; 2.66) among healthy males. In the TMD group, the corresponding values were 2.62 (95% CI: 2.21; 3.04) and 2.83 (95% CI: 1.67; 4.00), respectively. From the results of our study we conclude that moderate deviations from symmetric movements (mean: 1.2 mm for women, 2.1 mm for men) appear to be the norm even in healthy individuals. PMID- 15790378 TI - Comparison of ultrasonography and magnetic resonance imaging in the evaluation of temporomandibular joint disc displacement. AB - The aim of this work was to evaluate the accuracy and reliability of ultrasonography in the diagnosis of temporomandibular joint (TMJ) disc position abnormalities compared with magnetic resonance imaging (MRI). Participants in this study were 41 consecutive patients with signs and symptoms of temporomandibular disorders. All 82 TMJs were evaluated to detect disc position abnormalities by means of ultrasonography and MRI, performed by blinded operators. The accuracy of ultrasonography was evaluated with respect to MRI. Ultrasonography demonstrated good accuracy in the evaluation of disc position, showing a sensitivity of 65.8% and a specificity of 80.4%, resulting in a positive likelihood ratio of 3.35, a negative likelihood ratio of 0.42, and a diagnostic odds ratio of 7.97. The predictive positive and negatives values were respectively 77.1% and 70.2% and the overall agreement between the two radiological techniques was 73.1%. Ultrasonography proved to be accurate in detecting normal disc position and the presence of abnormalities in disc-condyle relationship but not so useful for the distinction between disc displacement with and without reduction. PMID- 15790379 TI - Type II collagen and aggrecan mRNA expressions in rabbit condyle following disc displacement. AB - The aim of this investigation was to study the remodelling of cartilage in the mandibular condyle following disc displacement (DD) of the temporomandibular joint (TMJ). Forty adult Japanese white rabbits were used in this study. The right joints of 28 of the 40 rabbits had their discs surgically displaced. Four of the 28 were killed at 4 days or 1, 2, 4, 6, 8 and 12 weeks after surgery. The messenger RNA (mRNA) expression levels of aggrecan and type II collagen in cartilages were measured using in situ hybridization techniques. Results showed that aggrecan mRNA expression reduced in the first week after DD. The expression began to recover after 4 weeks and reached a normal level after 6 weeks. Type II collagen mRNA expression reduced from 4 weeks and the expression recovered after 8 weeks. This suggests that the chondrocyte reacting to the displacement of the TMJ disc, alters its matrix gene expression patterns and it is may be the cause of the shape changes of TMJ after DD. PMID- 15790380 TI - Effect of luxation type on pulpal blood flow measurements: a long-term follow-up of luxated permanent maxillary incisors. AB - Laser Doppler flowmetry (LDF) is a non-invasive method to assess pulpal blood flow (PBF). Dental injury has been associated with significant PBF reduction. The purpose of this study was to assess whether (i) the type of luxation trauma may affect PBF measurements and (ii) whether luxation type-related measurements may show short- and long-term changes of PBF values. In 41 trauma patients, 69 maxillary incisor treated by repositioning and splinting, and the respective contralateral homologous tooth were investigated by LDF to assess local PBF values. Perfusion units were recorded in four sessions, on the day of splint removal, and 12, 24 and 36 weeks after splint removal. Statistical analysis consisted of univariate analysis of variance for repeated measurements. For the LDF measurements, the main effect of the variable 'session' was not significant (P = 0.119). However, there was a significant 'session'/'luxation type' interaction (P = 0.000). Analysis of simple session-within-luxation type effects revealed intrusive luxations to be associated with a significant decrease in PBF values (P = 0.000), while subluxations (P = 0.568), lateral luxations (P = 0.980), extrusive luxations (P = 0.910), and avulsions (P = 0.996) showed no significant difference between session-related values. The PBF measurements did not change over time for the contralateral incisors (P = 0.996). The LDF may become useful in the detection of pulpal ischaemic episodes in luxated maxillary incisors after repositioning and splinting. Further studies are warranted to assess the validity of the diagnosis of post-traumatic 'ischaemic episodes' by comparing it with histological tooth pulp changes, and by determining how well it may predict course and response to treatments in clinical trials. PMID- 15790381 TI - Survival rate and fracture strength of maxillary incisors, restored with different kinds of full veneers. AB - This in vitro study evaluated the survival rate and fracture strength of different kinds of ceramic full veneers, fabricated with a new experimental press ceramic (EPC VP2117/TC2, Ivoclar-Vivadent AG, Schaan, Liechtenstein). Eighty, caries-free human maxillary central incisors were used as abutments and were randomly divided into one control group and four test-groups of 16 samples each. In group A, unprepared teeth served as control. In the test groups, four different types of full veneer preparations were performed. In test groups B/C, the preparation was maintained in enamel and the contact point was on the tooth/on the veneer, respectively. In test groups D/E, the preparation was extended into dentin and the contact point was on the tooth/on the veneer, respectively. All veneers were adhesively luted using Variolink II (Ivoclar Vivadent AG). Then, the samples were exposed to the artificial mouth for 1.2 million chewing cycles (49 N). After exposure in the artificial mouth, a survival rate of the abutment teeth of 81-100% was reported among the different groups, but was not significantly different between the groups. However, no failures of the ceramic materials could be recognized. The median fracture strength of group A was 713.3 N, of group B 647.1 N, of group C 594.8 N, of group D 483.8 N and of group E 502.6 N. Among the different groups no significant difference was found. All mean values obtained were within the limits of clinical acceptance, indicating further clinical investigations on full veneers made out of the EPC. PMID- 15790382 TI - Effects of cantilever design and material on stress distribution in fixed partial dentures--a finite element analysis. AB - The purpose of this study was to examine the stress distribution in distal cantilevered fixed partial dentures (FPDs) that are designed with different cantilever morphology and made from different restorative materials. The finite element (FE) method was used to create models of two restoration types; metal ceramic and an all-ceramic FPDs. Both models were designed with distal cantilevers involving the first and second premolars as abutments and cantilever extension involving at the premolar or molar. The width of connector between the cantilever and the primary abutment restoration was 2.25 mm. The load applied during the FE analysis was positioned at the cusp tips of all teeth. The FE analysis of the models revealed that Von Mises stress values with maximum stress concentrations were observed on connectors of distal cantilevers. Stress concentration sites were also observed at the distal cervical area of the second premolar tooth. Models with premolar cantilever extensions restored with all ceramic induced lower Von Mises stress values than metal-ceramic restorations, however models with molar cantilever extensions restored with all-ceramic restorations induced higher Von Misses stress values than metal-ceramic restorations. If the distal cantilever length and restorative material is appropriately chosen, the failure frequency may be reduced. All ceramic can be used as restorative material, when the cantilevers length is not more than the mesiodistal dimension of a premolar tooth and metal-ceramic restorations can be used in longer situations. PMID- 15790383 TI - Influence of marginal bone resorption on stress around an implant--a three dimensional finite element analysis. AB - Average marginal bone resorption of about 1 mm after the first year of functional loading, which is followed by an annual loss of approximately 0.1 mm, has been reported in stable implants. However, finite element analyses on bone stress around implants have been limited to analysing the bone stress in the absence of any bone resorption. Thus, a three-dimensional finite element analysis was performed to compare the bone stresses in a non-resorption model with those in four models with bone resorption of two depths (1.3 and 2.6 mm) and types (horizontal resorption and angular defects). Axial and bucco-lingual forces were separately applied to the center of the superstructure and the maximum equivalent stress was calculated. The main tendencies of bone stress (highest stress concentration around implant neck, higher stresses under bucco-lingual than axial load, as well as in the cortical than cancellous bone) were the same in the non resorption and resorption models. Bone stress distributions were similar in the non-resorption and horizontal resorption models, but differed from those in the angular defect models. Moreover, the changes of the bone stress values with resorption depth differed for the two resorption types. Thus, in FEA, accurate simulation of the marginal bone shape in the implant neck region is advisable. PMID- 15790384 TI - Effects of salicylic-lactic acid conditioner on the shear bond strength of brackets and enamel surfaces. AB - The purpose of this study was to evaluate the effects of salicylic-lactic (SL) acid conditioner on the shear bond strength of brackets. Fluoride releasing (Light-bond) and non-fluoride releasing (Enlight) composite adhesives were used after conditioning with 0.22% salicylic + 9% lactic acid or 34% phosphoric acid. Composite adhesives were light cured with either a halogen light curing (HLC) unit for 30-50 s or a plasma arc curing (PAC) unit for 4 s. The shear bond strength was measured with an Instron. Failure modes of debonded brackets were identified based on adhesive remnants on the bracket and tooth. Salicylic-lactic acid conditioning was found to provide adequate shear bond strength. Groups conditioned with SL acid were debonded mainly at the enamel-resin interface and comparatively clean enamel surface after debonding was observed than those conditioned with phosphoric acid. Using confocal laser scanning microscopic examinations, it was found that demineralization patterns between SL acid and phosphoric acid conditioned groups were not different when the same adhesive was used. The SL acid conditioner did not reduce the demineralization. Light-bond adhesive showed less demineralization than Enlight adhesive. The PAC unit can be recommended as an alterative to the HLC unit because it significantly reduces the irradiation time. PMID- 15790385 TI - Bond strengths of nine current dentine adhesive systems to primary and permanent teeth. AB - The aim of this study was to compare the shear bond strength (SBS) of nine dentine bonding systems (DBS) of different classes to human primary and permanent dentine. Flat dentine occlusal surfaces were produced on human molars (100 primary, 100 permanent) by wet grinding on a 800-grit SiC paper. Nine DBS were applied following the manufacturers' instructions: One total etch multi-step system: Scotchbond Multipurpose Plus (3M/ESPE); Four total etch one-bottle system: Prime &Bond 2.1 (Dentsply), One Step (Bisco), Scotchbond 1 (3M/ESPE), and OptibondSolo Plus (Kerr); Three two-step self-etching primer systems: Clearfil Liner Bond 2 (Kuraray), Clearfil SE Bond (Kuraray), and Prime &Bond NT with NRC (Dentsply); An 'all-in-one' self-etching system: Prompt L-Pop (3M/ESPE). Composite (Z100; 3M/ESPE) cylinders (2 mm diameter, 3 mm high) were polymerized on the treated dentine surfaces and the specimens were stored at 37 degrees C for 24 h prior to testing. Twenty experimental groups were produced and tested. Statistical analysis revealed both a substrate and a bonding system effect. Two adhesive systems (One Step, Prime &Bond NT) had significantly higher bond strengths on permanent than on primary dentine. There was an effect of dentine bonding system on the mode of fracture. Although eight of the 10 DBS tested exhibited higher median SBS values on permanent dentine than on primary dentine, the dependent pairwise comparison identified a significant difference only for two groups. The use of simplified bonding systems does not necessarily result in improved bond strength to primary or to permanent dentine. PMID- 15790386 TI - A new method to measure the polymerization shrinkage kinetics of light cured composites. AB - This study was undertaken to develop a new measurement method to determine the initial dynamic volumetric shrinkage of composite resins during polymerization, and to investigate the effect of curing light intensity on the polymerization shrinkage kinetics. The instrument was basically an electromagnetic balance that was constructed with a force transducer using a position sensitive photo detector (PSPD) and a negative feedback servo amplifier. The volumetric change of composites during polymerization was detected continuously as a buoyancy change in distilled water by means of the Archimedes' principle. Using this new instrument, the dynamic patterns of the polymerization shrinkage of seven commercial composite resins were measured. The polymerization shrinkage of the composites was 1.92 approximately 4.05 volume %. The shrinkage of a packable composite was the lowest, and that of a flowable composite was the highest. The maximum rate of polymerization shrinkage increased with increasing light intensity but the peak shrinkage rate time decreased with increasing light intensity. A strong positive relationship was observed between the square root of the light intensity and the maximum shrinkage rate. The shrinkage rate per unit time, dVol%/dt, showed that the instrument can be a valuable research method for investigating the polymerization reaction kinetics. This new shrinkage-measuring instrument has some advantages that it was insensitive to temperature changes and could measure the dynamic volumetric shrinkage in real time without complicated processes. Therefore, it can be used to characterize the shrinkage kinetics in a wide range of commercial and experimental visible-light-cure materials in relation to their composition and chemistry. PMID- 15790387 TI - Accelerated probabilistic inference of RNA structure evolution. AB - BACKGROUND: Pairwise stochastic context-free grammars (Pair SCFGs) are powerful tools for evolutionary analysis of RNA, including simultaneous RNA sequence alignment and secondary structure prediction, but the associated algorithms are intensive in both CPU and memory usage. The same problem is faced by other RNA alignment-and-folding algorithms based on Sankoff's 1985 algorithm. It is therefore desirable to constrain such algorithms, by pre-processing the sequences and using this first pass to limit the range of structures and/or alignments that can be considered. RESULTS: We demonstrate how flexible classes of constraint can be imposed, greatly reducing the computational costs while maintaining a high quality of structural homology prediction. Any score-attributed context-free grammar (e.g. energy-based scoring schemes, or conditionally normalized Pair SCFGs) is amenable to this treatment. It is now possible to combine independent structural and alignment constraints of unprecedented general flexibility in Pair SCFG alignment algorithms. We outline several applications to the bioinformatics of RNA sequence and structure, including Waterman-Eggert N-best alignments and progressive multiple alignment. We evaluate the performance of the algorithm on test examples from the RFAM database. CONCLUSION: A program, Stemloc, that implements these algorithms for efficient RNA sequence alignment and structure prediction is available under the GNU General Public License. PMID- 15790388 TI - An entropy-based gene selection method for cancer classification using microarray data. AB - BACKGROUND: Accurate diagnosis of cancer subtypes remains a challenging problem. Building classifiers based on gene expression data is a promising approach; yet the selection of non-redundant but relevant genes is difficult. The selected gene set should be small enough to allow diagnosis even in regular clinical laboratories and ideally identify genes involved in cancer-specific regulatory pathways. Here an entropy-based method is proposed that selects genes related to the different cancer classes while at the same time reducing the redundancy among the genes. RESULTS: The present study identifies a subset of features by maximizing the relevance and minimizing the redundancy of the selected genes. A merit called normalized mutual information is employed to measure the relevance and the redundancy of the genes. In order to find a more representative subset of features, an iterative procedure is adopted that incorporates an initial clustering followed by data partitioning and the application of the algorithm to each of the partitions. A leave-one-out approach then selects the most commonly selected genes across all the different runs and the gene selection algorithm is applied again to pare down the list of selected genes until a minimal subset is obtained that gives a satisfactory accuracy of classification. The algorithm was applied to three different data sets and the results obtained were compared to work done by others using the same data sets. CONCLUSION: This study presents an entropy-based iterative algorithm for selecting genes from microarray data that are able to classify various cancer sub-types with high accuracy. In addition, the feature set obtained is very compact, that is, the redundancy between genes is reduced to a large extent. This implies that classifiers can be built with a smaller subset of genes. PMID- 15790389 TI - Smoking and tooth discolouration: findings from a national cross-sectional study. AB - BACKGROUND: Smoking is a risk factor of a number of oral diseases; the extent to which tobacco products influence dental aesthetics has not been widely investigated. The aim of this study was to determine satisfaction with own tooth colour of smokers and non-smokers and to investigate whether smokers have higher levels of self-assessed tooth discolouration compared to non-smokers METHODS: A cross sectional national study was conducted on sample of 6,000 UK adults. A total of 3,384 adults was interviewed. Smoking behaviour was recorded together with satisfaction with own tooth colour. Prevalence of perceived discolouration was measured by asking respondents to match their own tooth colour to one of a set of seven photographs of differing severities of discolouration. RESULTS: Twenty eight percent of smokers reported having moderate and severe levels of tooth discolouration compared to 15% in non-smokers. As well as more often perceiving discolouration smokers were also more likely to be dissatisfied with their own tooth colour compared to non-smokers. CONCLUSION: The study provides further evidence of the negative impact of tobacco smoking on dental aesthetics in the general public. The evidence provided by the study may be of value in short interventions for smoking cessation in the dental setting. PMID- 15790390 TI - Providing visualisation support for the analysis of anatomy ontology data. AB - BACKGROUND: Improvements in technology have been accompanied by the generation of large amounts of complex data. This same technology must be harnessed effectively if the knowledge stored within the data is to be retrieved. Storing data in ontologies aids its management; ontologies serve as controlled vocabularies that promote data exchange and re-use, improving analysis. The Edinburgh Mouse Atlas Project stores the developmental stages of the mouse embryo in anatomy ontologies. This project is looking at the use of visual data overviews for intuitive analysis of the ontology data. RESULTS: A prototype has been developed that visualises the ontologies using directed acyclic graphs in two dimensions, with the ability to study detail in regions of interest in isolation or within the context of the overview. This is followed by the development of a technique that layers individual anatomy ontologies in three-dimensional space, so that relationships across multiple data sets may be mapped using physical links drawn along the third axis. CONCLUSION: Usability evaluations of the applications confirmed advantages in visual analysis of complex data. This project will look next at data input from multiple sources, and continue to develop the techniques presented to provide intuitive identification of relationships that span multiple ontologies. PMID- 15790391 TI - First molecular screening of deafness in the Altai Republic population. AB - BACKGROUND: We studied the molecular basis of NSHL in Republic of Altai (South Siberia, Russia). The Altaians are the indigenous Asian population of the Altai Mountain region considered as a melting-pot and a dispersion center for world wide human expansions in the past. METHODS: A total of 76 patients of Altaian, Russian or mixed ethnicity and 130 Altaian controls were analyzed by PCR-DHPLC and sequencing in the GJB2 gene. The GJB6 deletion and the common non-syndromic deafness-causing mitochondrial mutations were also tested when appropriate. RESULTS: 8.3% of the Altaian chromosomes were carrying GJB2 mutations versus 46.9% of the Russian chromosomes. The 235delC mutation was predominant among Altaians, whereas the 35delG mutation was most prevalent among Russian patients. CONCLUSION: We found an Asian-specific GJB2 diversity among Altaians, and different GJB2 contribution for deafness in the Altaian and Russian patients. The high carrier frequency of 235delC in Altaians (4.6%) is probably defined by gene drift/founder effect in a particular group. The question whether the Altai region could be one of founder sources for the 235delC mutation widespread in Asia is open. PMID- 15790392 TI - Anticipatory nausea in cyclical vomiting. AB - BACKGROUND: Cyclical Vomiting Syndrome (CVS) is characterised by discrete, unexplained episodes of intense nausea and vomiting, and mainly affects children and adolescents. Comprehending Cyclical Vomiting Syndrome requires awareness of the severity of nausea experienced by patients. As a subjective symptom, nausea is easily overlooked, yet is the most distressing symptom for patients and causes many behavioural changes during attacks. CASE PRESENTATION: This first-hand account of one patient's experience of Cyclical Vomiting Syndrome shows how severe nausea contributed to the development of anticipatory nausea and vomiting (ANV), a conditioned response frequently observed in chemotherapy patients. This conditioning apparently worsened the course of the patient's disease. Anticipatory nausea and vomiting has not previously been recognised in Cyclical Vomiting Syndrome, however predictors of its occurrence in oncology patients indicate that it could complicate many cases. CONCLUSION: We suggest a model whereby untreated severe and prolonged nausea provokes anxiety about further cyclical vomiting attacks. This anxiety facilitates conditioning, thus increasing the range of triggers in a self-perpetuating manner. Effective management of the nausea-anxiety feedback loop can reduce the likelihood of anticipatory nausea and vomiting developing in other patients. PMID- 15790393 TI - Cloning and transcriptional analysis of the mouse receptor activity modifying protein-1 gene promoter. AB - BACKGROUND: Receptor activity modifying protein-1 (RAMP-1) is a single transmembrane-domain protein required for the functional expression of calcitonin gene-related peptide (CGRP) receptors. To date, little is known about the molecular mechanism(s) that activate/inhibit RAMP-1 gene expression. Such mechanism(s) are likely to play a major role in modulating the responsiveness of tissues to CGRP. RESULTS: To initiate studies on the transcriptional regulation of the mouse RAMP-1 gene, RAMP-1 transcriptional initiation sites were mapped in a variety of tissues. Analysis of RAMP-1 expression in C2C12 myoblasts demonstrated that RAMP-1 mRNA is expressed at greatest levels in confluent myoblasts verses non-confluent and fused myoblasts. Transfection of confluent C2C12 myoblasts and NIH 3T3 cells with RAMP-1 promoter/luciferase deletion constructs revealed that 4.7 kb of RAMP-1 5' flanking region demonstrated optimal promoter activity while 343 bp of 5' flanking region was defined as a minimal RAMP-1 promoter. In non-RAMP-1 expressing HEK293 cells, constructs containing 4.7 kb to 782 bp of RAMP-1 5' flanking region were transcriptionally inactive. However, deletion of sequences -782 to -343 activated RAMP-1 promoter activity. CONCLUSION: These findings suggest that tissue specificity of RAMP-1 gene expression is mediated by a negative acting transcription factor that represses RAMP-1 gene expression in non-RAMP-1 expressing tissues. This transcription factor is therefore likely to play an important role in modulating the responsiveness of tissues to CGRP. PMID- 15790394 TI - Glycogen synthesis correlates with androgen-dependent growth arrest in prostate cancer. AB - BACKGROUND: Androgen withdrawal in normal prostate or androgen-dependent prostate cancer is associated with the downregulation of several glycolytic enzymes and with reduced glucose uptake. Although glycogen metabolism is known to regulate the intracellular glucose level its involvement in androgen response has not been studied. METHODS: We investigated the effects of androgen on glycogen phosphorylase (GP), glycogen synthase (GS) and on glycogen accumulation in the androgen-receptor (AR) reconstituted PC3 cell line containing either an empty vector (PC3-AR-V) or vector with HPV-E7 (PC3-AR-E7) and the LNCaP cell line. RESULTS: Androgen addition in PC3 cells expressing the AR mimics androgen ablation in androgen-dependent prostate cells. Incubation of PC3-AR-V or PC3-AR E7 cells with the androgen R1881 induced G1 cell cycle arrest within 24 hours and resulted in a gradual cell number reduction over 5 days thereafter, which was accompanied by a 2 to 5 fold increase in glycogen content. 24 hours after androgen-treatment the level of Glucose-6-P (G-6-P) had increased threefold and after 48 hours the GS and GP activities increased twofold. Under this condition inhibition of glycogenolysis with the selective GP inhibitor CP-91149 enhanced the increase in glycogen content and further reduced the cell number. The androgen-dependent LNCaP cells that endogenously express AR responded to androgen withdrawal with growth arrest and increased glycogen content. CP-91149 further increased glycogen content and caused a reduction of cell number. CONCLUSION: Increased glycogenesis is part of the androgen receptor-mediated cellular response and blockage of glycogenolysis by the GP inhibitor CP-91149 further increased glycogenesis. The combined use of a GP inhibitor with hormone therapy may increase the efficacy of hormone treatment by decreasing the survival of prostate cancer cells and thereby reducing the chance of cancer recurrence. PMID- 15790395 TI - Acne and smoking: is there a relationship? AB - BACKGROUND: There are contradictory reports on the relationship between acne vulgaris and cigarette smoking. The objective of this study was to examine the relation between acne and cigarette smoking in a case-control study. METHODS: A questionnaire on smoking habits was offered to 350 patients with acne vulgaris and 350 patients suffering from skin diseases other than acne, aged 15-40 years, attending in a skin clinic in Tehran, Iran. The patients completed the questionnaires anonymously in the waiting room. RESULTS: Two hundred and ninety three patients with acne (response rate 83.7 %) and 301 patients with other skin diseases (response rate 86.0 %) completed the questionnaires. Twelve acne patients (4.1 %) and 27 control patients (9.0 %) were current smokers (odds ratio = 0.43, 95% confidence limits 0.22 - 0.87, p < 0.05). But after adjustment for sex, this difference was not significant (odds ratio: 0.61, 95% CI: 0.30-1.26, p > 0.05, Mantel-Haenszel test). CONCLUSION: An association between acne and cigarette smoking was not found in this study. PMID- 15790396 TI - Paircomp, FamilyRelationsII and Cartwheel: tools for interspecific sequence comparison. AB - BACKGROUND: Comparative sequence analysis is an effective and increasingly common way to identify cis-regulatory regions in animal genomes. RESULTS: We describe three tools for comparative analysis of pairs of BAC-sized genomic regions. Paircomp is a tool that does windowed (ungapped) comparisons of two sequences and reports all matches above a set threshold. FamilyRelationsII is a graphical viewer for comparisons that enables interactive exploration of several different kinds of comparisons. Cartwheel is a Web site and compute-cluster management system used to execute and store comparisons for display by FamilyRelationsII. These tools are specialized for the discovery of cis-regulatory regions in animal genomes. All tools and their source code are freely available at http://family.caltech.edu/. CONCLUSION: These tools have been shown to effectively identify regulatory regions in echinoderms, mammals, and nematodes. PMID- 15790398 TI - Genomic organization, annotation, and ligand-receptor inferences of chicken chemokines and chemokine receptor genes based on comparative genomics. AB - BACKGROUND: Chemokines and their receptors play important roles in host defense, organogenesis, hematopoiesis, and neuronal communication. Forty-two chemokines and 19 cognate receptors have been found in the human genome. Prior to this report, only 11 chicken chemokines and 7 receptors had been reported. The objectives of this study were to systematically identify chicken chemokines and their cognate receptor genes in the chicken genome and to annotate these genes and ligand-receptor binding by a comparative genomics approach. RESULTS: Twenty three chemokine and 14 chemokine receptor genes were identified in the chicken genome. All of the chicken chemokines contained a conserved CC, CXC, CX3C, or XC motif, whereas all the chemokine receptors had seven conserved transmembrane helices, four extracellular domains with a conserved cysteine, and a conserved DRYLAIV sequence in the second intracellular domain. The number of coding exons in these genes and the syntenies are highly conserved between human, mouse, and chicken although the amino acid sequence homologies are generally low between mammalian and chicken chemokines. Chicken genes were named with the systematic nomenclature used in humans and mice based on phylogeny, synteny, and sequence homology. CONCLUSION: The independent nomenclature of chicken chemokines and chemokine receptors suggests that the chicken may have ligand-receptor pairings similar to mammals. All identified chicken chemokines and their cognate receptors were identified in the chicken genome except CCR9, whose ligand was not identified in this study. The organization of these genes suggests that there were a substantial number of these genes present before divergence between aves and mammals and more gene duplications of CC, CXC, CCR, and CXCR subfamilies in mammals than in aves after the divergence. PMID- 15790399 TI - A simplified method for analysis of polyunsaturated fatty acids. AB - BACKGROUND: Analysis of fatty acid composition of biological materials is a common task in lipid research. Conventionally, preparation of samples for fatty acid analysis by gas chromatography involves two separate procedures: lipid extraction and methylation. This conventional method is complicated, tedious and time consuming. Development of a rapid and simple method for lipid analysis is warranted. RESULTS: We simplified the conventional method by combining the extraction and methylation into a single step (omitting the procedure of prior extraction). Various biological samples including cultured cells, animal tissues and human specimens have been tested using the new method. Statistical analysis indicates that the recovery of long chain fatty acids from tissue samples by the simplified method is significantly higher than that by the traditional method, but there is no difference in relative fatty acid composition between the two methods. This simplified method can significantly save time and materials, and reduce the potentials of sample loss and contamination. CONCLUSION: The lipid extraction procedure prior to methylation employed conventionally in lipid analysis can be omitted without affecting the recovery of long chain (>or= 18 C) fatty acids and their composition. The simplified method is rapid, easy-to-use, suitable for analysis of total long chain polyunsaturated fatty acid contents (e.g. n-6 and n-3 fatty acids) in various biological samples, especially when the number of samples to be analyzed is large and/or the specimen size is small. PMID- 15790397 TI - NXT2 is required for embryonic heart development in zebrafish. AB - BACKGROUND: NXT2 is a member of NXT family proteins that are generally involved in exporting nuclear RNA in eukaryotic cells. It is not known if NXT2 has any function in specific biological processes. RESULTS: A zebrafish mutant exhibiting specific heart defects during embryogenesis was generated by animal cloning mediated retroviral insertions. Molecular analysis indicated that the mutant phenotype was caused by a disruption of NXT2. Whole-mount RNA in situ hybridization showed that NXT2 transcripts were clearly detectable in embryonic heart as well as other tissues. Further analysis revealed that expression level of one form of alternative splicing NXT2 mRNA transcripts was significantly reduced, resulting in deficient myocardial cell differentiation and the malformation of cardiac valve at the atrioventricular boundary. The defects could be reproduced by morpholino anti-sense oligo knockdown of NXT2. CONCLUSION: NXT2 has a critical role in maintaining morphogenetic integrity of embryonic heart in vertebrate species. PMID- 15790400 TI - Muscle oxygenation trends after tapering in trained cyclists. AB - BACKGROUND: This study examined muscle deoxygenation trends before and after a 7 day taper using non-invasive near infrared spectroscopy (NIRS). METHODS: Eleven cyclists performed an incremental cycle ergometer test to determine maximal oxygen consumption (VO2max = 4.68 +/- 0.57 L.min-1) prior to the study, and then completed two or three high intensity (85-90% VO2max) taper protocols after being randomly assigned to a taper group: T30 (n = 5), T50 (n = 5), or T80 (n = 5) [30%, 50%, 80% reduction in training volume, respectively]. Physiological measurements were recorded during a simulated 20 km time trials (20TT) performed on a set of wind-loaded rollers. RESULTS AND DISCUSSION: The results showed that the physiological variables of oxygen consumption (VO2), carbon dioxide (VCO2) and heart rate (HR) were not significantly different after tapering, except for a decreased ventilatory equivalent for oxygen (VE/VO2) in T50 (p or = 65) by Fisher exact test and exact Wilcoxon rank-sum test. RESULTS: From March 1991 to March 2002, 180 patients were identified, 100 older than 60 and younger than 65, and 80 aged 65 or older. Febrile neutropenia was more frequent among older patients (p = 0.05). Leukopenia, neutropenia, nausea, cardiac toxicity and thrombophlebitis tended to be more frequent or severe among elderlies, while mucositis tended to be more evident among younger patients, all not significantly. Almost one half (47%) of the older patients receiving concomitant radiotherapy experienced grade 3-4 haematological toxicity. Compliance was similar in the two groups, with 6 cycles administered in 86% and 79%, day-8 chemotherapy omitted at least once in 36% and 39%, dose reduction in 27% and 38%, prolonged treatment duration (> or = 29 weeks) in 10% and 11% and need of G-CSF in 9% and 18%, among younger and older patients, respectively. CONCLUSION: Our data show that, in a highly selected population of patients 65 or more years old, CMF is as feasible as in patients older than 60 and younger than 65, but with a relevant burden of toxicity. We suggest that prospective trials in elderly patients testing less toxic treatment schemes are mandatory before indicating adjuvant chemotherapy to all elderly patients with significant risk of breast cancer recurrence. PMID- 15790418 TI - Prediction of a common structural scaffold for proteasome lid, COP9-signalosome and eIF3 complexes. AB - BACKGROUND: The 'lid' subcomplex of the 26S proteasome and the COP9 signalosome (CSN complex) share a common architecture consisting of six subunits harbouring a so-called PCI domain (proteasome, CSN, eIF3) at their C-terminus, plus two subunits containing MPN domains (Mpr1/Pad1 N-terminal). The translation initiation complex eIF3 also contains PCI- and MPN-domain proteins, but seems to deviate from the 6+2 stoichiometry. Initially, the PCI domain was defined as the region of detectable sequence similarity between the components mentioned above. RESULTS: During an exhaustive bioinformatical analysis of proteasome components, we detected multiple instances of tetratrico-peptide repeats (TPR) in the N terminal region of most PCI proteins, suggesting that their homology is not restricted to the PCI domain. We also detected a previously unrecognized PCI domain in the eIF3 component eIF3k, a protein whose 3D-structure has been determined recently. By using profile-guided alignment techniques, we show that the structural elements found in eIF3k are most likely conserved in all PCI proteins, resulting in a structural model for the canonical PCI domain. CONCLUSION: Our model predicts that the homology domain PCI is not a true domain in the structural sense but rather consists of two subdomains: a C-terminal 'winged helix' domain with a key role in PCI:PCI interaction, preceded by a helical repeat region. The TPR-like repeats detected in the N-terminal region of PCI proteins most likely form an uninterrupted extension of the repeats found within the PCI domain boundaries. This model allows an interpretation of several puzzling experimental results. PMID- 15790419 TI - Effects of first and second generation antihistamines on muscarinic induced mucus gland cell ion transport. AB - BACKGROUND: The first generation antihistamines, such as diphenhydramine, are fairly potent muscarinic antagonists in addition to being H1 selective antihistamines. The antimuscarinic action is often not desirable since it is in part responsible for the drying of secretions in the airways and the sedative effect. We therefore examined a number of antihistamines for antimuscarinic effects on ion transport by mucus gland cells isolated from the airways of swine. Enzymatically isolated airway mucus gland cells were purified utilizing density gradients and grown in culture on porous inserts (Millicell HA) at an air interface. Cells grown in this manner maintain phenotype and polarity. Transport of ions, as short-circuit current measured under voltage-clamp, was measured in response to acetylcholine (ACh) or histamine applied to the serosal side of the gland cell layers. Concentration-response relationships for ACh or histamine were generated in the presence and absence of various drugs. The potencies against muscarinic receptor activation were estimated using the dose-ratio method of Schild. RESULTS: Three known muscarinic antagonists were used to validate the system. Atropine had a pA2 of 9.4 +/- 0.1 (n = 9). 4-DAMP and methoctramine had pA2 values of 8.6 +/- 0.1 and 5.6 +/- 0.1, respectively (n = 12, 11) all consistent with inhibition of an M3 subtype muscarinic receptor. The rank order of potency of the antihistamines against the inhibition of M3 receptors was desloratadine = diphenhydramine > hydroxyzine (pA2; 6.4, 6.2, 4.8, respectively). pA2 values for fexofenadine, loratadine and cetirizine were not determined since they had no effect on the cholinergic response at the highest drug concentrations tested (10, 10 and 100 microM, respectively). The pA2 values for the antihistamines against the histamine response could not be calculated, but the estimates of the rank order of potency were estimated to be desloratadine > cetirizine approximate to hydroxyzine > fexofenadine > loratadine > diphenhydramine. CONCLUSION: The rank order of selectivity for histamine receptors over muscarinic receptors was estimated to be cetirizine approximate to fexofenadine > loratadine > desloratadine > or = hydroxyzine > or = diphenhydramine. PMID- 15790420 TI - The relation between plasma tyrosine concentration and fatigue in primary biliary cirrhosis and primary sclerosing cholangitis. AB - BACKGROUND: In primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) fatigue is a major clinical problem. Abnormal amino acid (AA) patterns have been implicated in the development of fatigue in several non-hepatological conditions but for PBC and PSC no data are available. This study aimed to identify abnormalities in AA patterns and to define their relation with fatigue. METHODS: Plasma concentrations of tyrosine, tryptophan, phenylalanine, valine, leucine and isoleucine were determined in plasma of patients with PBC (n = 45), PSC (n = 27), chronic hepatitis C (n = 22) and healthy controls (n = 73). Fatigue and quality of life were quantified using the Fisk fatigue severity scale, a visual analogue scale and the SF-36. RESULTS: Valine, isoleucine, leucine were significantly decreased in PBC and PSC. Tyrosine and phenylalanine were increased (p < 0.0002) and tryptophan decreased (p < 0.0001) in PBC. In PBC, but not in PSC, a significant inverse relation between tyrosine concentrations and fatigue and quality of life was found. Patients without fatigue and with good quality of life had increased tyrosine concentrations compared to fatigued patients. Multivariate analysis indicated that this relation was independent from disease activity or severity or presence of cirrhosis. CONCLUSION: In patients with PBC and PSC, marked abnormalities in plasma AA patterns occur. Normal tyrosine concentrations, compared to increased concentrations, may be associated with fatigue and diminished quality of life. PMID- 15790421 TI - Ranking the whole MEDLINE database according to a large training set using text indexing. AB - BACKGROUND: The MEDLINE database contains over 12 million references to scientific literature, with about 3/4 of recent articles including an abstract of the publication. Retrieval of entries using queries with keywords is useful for human users that need to obtain small selections. However, particular analyses of the literature or database developments may need the complete ranking of all the references in the MEDLINE database as to their relevance to a topic of interest. This report describes a method that does this ranking using the differences in word content between MEDLINE entries related to a topic and the whole of MEDLINE, in a computational time appropriate for an article search query engine. RESULTS: We tested the capabilities of our system to retrieve MEDLINE references which are relevant to the subject of stem cells. We took advantage of the existing annotation of references with terms from the MeSH hierarchical vocabulary (Medical Subject Headings, developed at the National Library of Medicine). A training set of 81,416 references was constructed by selecting entries annotated with the MeSH term stem cells or some child in its sub tree. Frequencies of all nouns, verbs, and adjectives in the training set were computed and the ratios of word frequencies in the training set to those in the entire MEDLINE were used to score references. Self-consistency of the algorithm, benchmarked with a test set containing the training set and an equal number of references randomly selected from MEDLINE was better using nouns (79%) than adjectives (73%) or verbs (70%). The evaluation of the system with 6,923 references not used for training, containing 204 articles relevant to stem cells according to a human expert, indicated a recall of 65% for a precision of 65%. CONCLUSION: This strategy appears to be useful for predicting the relevance of MEDLINE references to a given concept. The method is simple and can be used with any user-defined training set. Choice of the part of speech of the words used for classification has important effects on performance. Lists of words, scripts, and additional information are available from the web address http://www.ogic.ca/projects/ks2004/. PMID- 15790422 TI - Exercise responsive genes measured in peripheral blood of women with chronic fatigue syndrome and matched control subjects. AB - BACKGROUND: Chronic fatigue syndrome (CFS) is defined by debilitating fatigue that is exacerbated by physical or mental exertion. To search for markers of CFS associated post-exertional fatigue, we measured peripheral blood gene expression profiles of women with CFS and matched controls before and after exercise challenge. RESULTS: Women with CFS and healthy, age-matched, sedentary controls were exercised on a stationary bicycle at 70% of their predicted maximum workload. Blood was obtained before and after the challenge, total RNA was extracted from mononuclear cells, and signal intensity of the labeled cDNA hybridized to a 3800-gene oligonucleotide microarray was measured. We identified differences in gene expression among and between subject groups before and after exercise challenge and evaluated differences in terms of Gene Ontology categories. Exercise-responsive genes differed between CFS patients and controls. These were in genes classified in chromatin and nucleosome assembly, cytoplasmic vesicles, membrane transport, and G protein-coupled receptor ontologies. Differences in ion transport and ion channel activity were evident at baseline and were exaggerated after exercise, as evidenced by greater numbers of differentially expressed genes in these molecular functions. CONCLUSION: These results highlight the potential use of an exercise challenge combined with microarray gene expression analysis in identifying gene ontologies associated with CFS. PMID- 15790425 TI - Screening for novel enzymes from metagenome and SIGEX, as a way to improve it. AB - Metagenomics has been successfully applied to isolate novel biocatalysts from the uncultured microbiota in the environment. Two types of screening have been used to identify clones carrying desired traits from metagenomic libraries: function based screening, and sequence-based screening. Both function- and sequence- based screening have individual advantages and disadvantages, and they have been applied successfully to discover biocatalysts from metagenome. However, both strategies are laborious and tedious because of the low frequency of screening hits. A recent paper introduced a high throughput screening strategy, termed substrate-induced gene-expression screening (SIGEX). SIGEX is designed to select the clones harboring catabolic genes induced by various substrates in concert with fluorescence activated cell sorting (FACS). This method was applied successfully to isolate aromatic hydrocarbon-induced genes from a metagenomic library. Although SIGEX has many limitations, it is expected to provide economic advantages, especially to industry. PMID- 15790424 TI - Two dimensional VOPBA reveals laminin receptor (LAMR1) interaction with dengue virus serotypes 1, 2 and 3. AB - BACKGROUND: The search for the dengue virus receptor has generated many candidates often identified only by molecular mass. The wide host range of the viruses in vitro combined with multiple approaches to identifying the receptor(s) has led to the notion that many receptors or attachment proteins may be involved and that the different dengue virus serotypes may utilize different receptors on the same cells as well as on different cell types. RESULTS: In this study we used sequential extraction of PS Clone D cell monolayers with the detergent beta octylglucopyranoside followed by sodium deoxycholate to prepare a cell membrane rich fraction. We then used 2 dimensional (2D) gel electrophoresis to separate the membrane proteins and applied a modified virus overlay protein binding assay (VOPBA) to show that dengue virus serotypes 1, 2 and 3 all interact with the 37 kDa/67 kDa laminin receptor (LAMR1), a common non-integrin surface protein on many cell types. CONCLUSION: At least 3 of the 4 dengue serotypes interact with the 37 kDa/67 kDa laminin receptor, LAMR1, which may be a common player in dengue virus-cell surface interaction. PMID- 15790423 TI - Effects of Maharishi Amrit Kalash 5 as an Ayurvedic herbal food supplement on immune functions in aged mice. AB - BACKGROUND: Maharishi Amrit Kalash (MAK) 5, one of the Ayurvedic food supplements, belongs to a group of substances known as Rasayana. MAK5 and other Rasayanas are believed to enhance the body's resistance to infections and disease, and enhance longevity. In this study, we determined the effects of administration of MAK5, one of the Ayurvedic food supplements on immune functions in young and old mice. METHODS: Male C3H/He N mice were divided into five groups: two no treatment groups (old control: 22-month-old and young control: 2-month old) and three MAK5 treated groups with differing dose of MAK5. MAK5 was given p.o. at 50 mg/kg, 100 mg/kg or 200 mg/kg per day (3 days/week) for 2 months. RESULTS: We found that glucose consumption of peritoneal macrophages from old mice treated with MAK5 at all doses and incubated for 48 and 72 h were significantly greater than that in the control group. Nitric oxide production of peritoneal macrophages stimulated by lipopolysaccharide (LPS) in old mice treated with MAK5 at all doses was significantly greater than that in the old control group, but not compared to the young control group. Stimulation index (S.I.) in old mice gavaged with MAK5 at all doses was significantly higher than that in the old control group. IL-2 production stimulated by Con A in old mice given MAK5 at all doses was significantly higher than that in the old control group. Production of IFN-gamma stimulated by Con A in old mice given MAK5 at doses of 100 mg/kg and 200 mg/kg were significantly higher than that in the old control group. IL-4 production of splenic lymphocyte stimulated by Con A in old mice given MAK5 at dose levels of 100 and 200 mg/kg were significantly higher than that in the old control group. CONCLUSION: The results suggest that MAK5 suppressed the age associated glucose consumption of peritoneal macrophages and cellular immune function reduction, and that it contributes to the prevention of the immunosenescence. PMID- 15790426 TI - Tyrosine phosphorylation of myosin heavy chain during skeletal muscle differentiation: an integrated bioinformatics approach. AB - BACKGROUND: Previously it has been shown that insulin-mediated tyrosine phosphorylation of myosin heavy chain is concomitant with enhanced association of C-terminal SRC kinase during skeletal muscle differentiation. We sought to identify putative site(s) for this phosphorylation event. RESULTS: A combined bioinformatics approach of motif prediction and evolutionary and structural analyses identified tyrosines163 and 1856 of the skeletal muscle heavy chain as the leading candidate for the sites of insulin-mediated tyrosine phosphorylation. CONCLUSION: Our work is suggestive that tyrosine phosphorylation of myosin heavy chain, whether in skeletal muscle or in platelets, is a significant event that may initiate cytoskeletal reorganization of muscle cells and platelets. Our studies provide a good starting point for further functional analysis of MHC phosphor-signalling events within different cells. PMID- 15790427 TI - Comparative mapping of sequence-based and structure-based protein domains. AB - BACKGROUND: Protein domains have long been an ill-defined concept in biology. They are generally described as autonomous folding units with evolutionary and functional independence. Both structure-based and sequence-based domain definitions have been widely used. But whether these types of models alone can capture all essential features of domains is still an open question. METHODS: Here we provide insight on domain definitions through comparative mapping of two domain classification databases, one sequence-based (Pfam) and the other structure-based (SCOP). A mapping score is defined to indicate the significance of the mapping, and the properties of the mapping matrices are studied. RESULTS: The mapping results show a general agreement between the two databases, as well as many interesting areas of disagreement. In the cases of disagreement, the functional and evolutionary characteristics of the domains are examined to determine which domain definition is biologically more informative. PMID- 15790429 TI - CO(2) pneumoperitoneum or the Bookwalter: choose your access and exposure. PMID- 15790428 TI - Osteo-Promoter Database (OPD) -- promoter analysis in skeletal cells. AB - BACKGROUND: Increasing our knowledge about the complex expression of genes in skeletal tissue will provide a better understanding of the physiology of skeletal cells. The study summarizes transcriptional regulation factors interacting and cooperating at promoter regions that regulate gene expression. Specifically, we analyzed A/T rich elements along the promoter sequences. DESCRIPTION: The Osteo Promoter Database (OPD) is a collection of genes and promoters expressed in skeletal cells. We have compiled a new viewer, OPD, as unique database developed and created as an accessible tool for skeletal promoter sequences. OPD can navigate to identify genes specific to skeletal cDNA databases and promoter analysis sites. OPD offers exclusive access to facilitate a dynamic extraction of promoters' gene-specific analyses in skeletal tissue. The data on promoters included in OPD contains cloned promoters or predicted promoters that were analyzed by bioinformatics tools. OPD offers MAR-analysis, which allocates A/T rich elements along these promoter sequences. CONCLUSION: The analysis leads to a better insight of proteins that bind to DNA, regulate DNA, and function in chromatin remodeling. The OPD is a distinctive tool for understanding the complex function of chromatin remodeling and transcriptional regulation of specific gene expression in skeletal tissue. PMID- 15790430 TI - Analysis of morbidity in patients with endometrial cancer: is there a commitment to offer laparoscopy? AB - OBJECTIVE: Benefits of laparoscopy over laparotomy in patients with endometrial cancer (EC) are well known. As many patients with EC carry co-morbid conditions, surgery is exposing them to increased risk of complications. A review of the patients with EC recruited so far in a clinical trial comparing laparoscopy to laparotomy was performed. The goal was to identify patients carrying specific risk factors for complications, who would most benefit of laparoscopy and be the ideal candidates for this surgical approach. PATIENTS AND METHODS: Between July 1995 and December 2002, 122 patients with uterine cancer entered the study. Sixty three patients were allocated to the laparoscopy (LPS) arm (group A), while 59 were allocated to the laparotomy (LPT) arm (group B). Rate and type of intra-, early and late post-operative complications were prospectively recorded. Risk factors for complications are analyzed to define a group of patients truly benefiting from laparoscopy. RESULTS: Overall, 12 patients out of 122 (9.8%) have experienced intra-operative, 43 patients out of 122 (35.2%) early post-operative and 25 patients out of 122 (20.4%) late post-operative complications. Rate of intra-operative complications was 4.7% in group A (3 patients out of 63) vs. 15.2% in group B (9 patients out of 59), P = 0.082. Early post-operative complications rate was 23.8% in group A (15 out of 63) and 47.4% in group B (28 out of 59), P = 0.011. Rate of late post-operative complications was 7.9% (5 out of 63) in group A vs. 35.5% (21 out of 59), P = 0.001. Univariate analysis shows co-morbid medical conditions, weight >80 kg, Quetelet index >30 and age >65 years to be predictive of complications and, in fact, a subgroup of patients presenting with these characteristics (n = 57, 30 in group A and 27 in group B) has been recognized to accumulate 60% of the overall complications. In these patients, multivariate analysis identifies the surgical technique (LPS vs. LPT) to be the only significant risk factor for complications. CONCLUSION: At least one third of the patients with EC carry serious co-morbidities with an increased surgical risk for complications. For this subgroup of patients, a laparoscopic-vaginal approach significantly reduces the rate of complications and should be the standard of surgical treatment. PMID- 15790431 TI - Intraperitoneal carboplatin-based chemotherapy for epithelial ovarian cancer. AB - OBJECTIVE: Intraperitoneal (IP) chemotherapy is an attractive approach to the treatment of ovarian cancer because the disease remains confined to the peritoneal cavity for a large part of its natural history. In this review article, we discuss the current status and future perspectives of IP chemotherapy for ovarian cancer. METHODS: A systematic review of IP carboplatin-based chemotherapy for ovarian cancer treatment was conducted through evaluation of published manuscripts as well as abstracts from the proceedings of the American Society of Clinical Oncology and the Society of Gynecologic Oncologists. RESULTS: Despite the fact that survival or progression-free survival benefit has been demonstrated by three large randomized trials (Survival Hazard Ratio for GOG104/SWOG 0.76, Relative Risk Reduction of Progression: 0.78 for GOG114, and 0.73 for GOG172), IP chemotherapy has not been accepted as a standard treatment. This appears to be mainly due to the cisplatin-related toxicity and IP catheter failure. Substituting cisplatin with carboplatin may make this approach more tolerable. Recent pharmacological data suggest that carboplatin-based IP chemotherapy not only exposes the peritoneal cavity to higher doses of the agent, but also attains levels of systemic platinum comparable to intravenous (IV) administration. A large retrospective study has demonstrated excellent survival for patients with advanced ovarian cancer who underwent carboplatin-based IP chemotherapy. Low observed toxicity as well as infrequent catheter-related complications was significant findings in this report. CONCLUSION: A large scale phase III randomized trial is now urgently needed to clarify the role of carboplatin-based IP chemotherapy in ovarian cancer. PMID- 15790432 TI - Expression of steroid hormone receptors in BRCA1-associated ovarian carcinomas. AB - OBJECTIVE: BRCA1 mutations predispose to cancer in hormone responsive tissues. A predominance of estrogen receptor (ER)-negative breast cancers in BRCA1 mutation carriers and potential interactions between ERalpha and BRCA1 suggest a link between hormones and BRCA1. However, the expression pattern of ERalpha and other hormone receptors in BRCA1-associated ovarian cancer was unknown. METHODS: Twenty two BRCA1-associated ovarian cancer cases were matched with sporadic cases (no family history of ovarian or breast cancer) for FIGO stage, grade, histologic subtype, and patient age and hormone receptor expression was measured immunohistochemically. RESULTS: ERalpha expression was similar in BRCA1 associated ovarian cancer compared with matched sporadic counterparts, in contrast with previous findings in BRCA1-linked breast cancer. There was also no significant difference in expression of progesterone receptors and androgen receptor between the matched cases in the two groups. However, differences were noted in the relative expression of receptor isotypes, in particular, levels of ERalpha and ERbeta were positively correlated in sporadic tumors but inversely related in BRCA1-associated tumors. CONCLUSION: Similar hormone receptor expression in BRCA1-associated ovarian cancer and matched sporadic counterparts may be further evidence that at least a proportion of sporadic ovarian tumors and BRCA1-associated tumors develop through similar pathways. PMID- 15790433 TI - Activation of PI3K/Akt pathway by PTEN reduction and PIK3CA mRNA amplification contributes to cisplatin resistance in an ovarian cancer cell line. AB - OBJECTIVE: The aim of this study was to understand the role of PIK3CA and PTEN on the resistance of human ovarian cancer cells to cisplatin-induced apoptosis. METHODS: Human ovarian cancer cell OVCAR-3 and cisplatin-resistant subclone OVCAR 3/CDDP cells were used for these studies. The expressions of apoptosis regulating proteins and PI3K/Akt signaling proteins were systematically examined. RESULTS: OVCAR-3/CDDP cells were 4.8-fold more resistant to cisplatin compared to OVCAR-3 cells following 72 h exposure to this drug. This resistance was paralleled with reduced susceptibility to cisplatin-induced apoptosis. Apoptotic proteins were differentially expressed in OVCAR-3/CDDP cells, resulting in the inhibition of Bax translocalization. Cisplatin inhibited Akt phosphorylation and activation in OVCAR-3 cells but not in OVCAR-3/CDDP cells. The specific PI3K inhibitors LY294002 and wortmannin sensitized OVCAR-3/CDDP cells to cisplatin-induced apoptosis and decreased cell viability. A low level of PTEN expression was strongly associated with amplified PIK3CA and PI3K/Akt activities in OVCAR-3/CDDP cells. Small interfering RNA knockdown of PTEN and the expression of active p110alpha resulted in a blockade of apoptosis by cisplatin in OVCAR-3 cells. CONCLUSIONS: These results collectively indicate that the development of resistance in OVCAR-3 cells was derived by increased PIK3CA transcription and reduction of PTEN expression. These alterations conferred cisplatin resistance to cisplatin through the activation of PI3K/Akt and the inhibition of Bax translocation. PMID- 15790434 TI - Is p16(INK4A) expression more useful than human papillomavirus test to determine the outcome of atypical squamous cells of undetermined significance-categorized Pap smear? A comparative analysis using abnormal cervical smears with follow-up biopsies. AB - OBJECTIVE: To correlate high-risk human papillomavirus (HR-HPV) viral load to p16(INK4A) expression in atypical squamous cells of undetermined significance (ASCUS)-categorized Pap smears with follow-up biopsies in order to elucidate their relationships in gynecological pathology. METHODS: We studied 66 ASCUS categorized Pap smears with subsequent follow-up biopsies. HR-HPV viral load was determined by Hybrid Capture II assay from the cervical swab in each ASCUS diagnosed Pap smear. Both smears and biopsies were immunostained with a primary anti-p16 antibody, clone E6H4, and we analyzed the correlations between HR-HPV viral load in each ASCUS-diagnosed Pap smear and p16 expression of smears with follow-up biopsies. RESULTS: Correlation analyses of the corresponding histological diagnoses from 66 ASCUS-diagnosed Pap smears revealed that 21 (32%) cases had only reactive changes and 45 cases showed cervical intraepithelial neoplasia including LSIL (24 cases, 36%) and HSIL or higher (21 cases, 32%). Tests for HR-HPV viral load revealed 17 (26%) negative cases and 49 (74%) positive cases. Immunostaining showed that 26 cases (39%) were negative and 40 (61%) were positive for p16 expression. Comparative analysis of these two tests indicated consistencies as well as discrepancies. They showed significant differences (P < 0.001) between negative p16 expressions of Pap smears with the presence of reactive lesions in follow-up biopsies and HR-HPV viral load. However, no significant difference (P = 0.739 and 0.606) between p16 expression of Pap smears with the presence of LSIL, HSIL or higher in follow-up biopsies and high HR-HPV viral load was found. In addition, there were significant differences (P < 0.001) in specificity and positive predictive value, but no significant differences were found in sensitivity (P = 0.606) and negative predictive value (P = 0.062) between p16 immunostaining and HR-HPV viral load. CONCLUSION: The results indicate that there is a close association between weak or strong p16 expressions in ASCUS-categorized smears with the presence of SILs in follow-up biopsies and positive HR-HPV viral loads. Conversely, there is also a clear association between the lack of p16 expression and the absence of significant lesions in follow-up biopsies, but this is not consistent with a negative HR-HPV viral load. It is concluded that p16 expression is an indicator of pathogenic activity of HR-HPV, which is an objective biomarker for clarification of ASCUS categorized Pap smears in gynecological cytopathology. Furthermore, through comparative analysis, directly visualized p16 immunostaining on smears appears to be a more effective method than HR-HPV viral load for the detection of reactive changes and LSILs from ASCUS-categorized Pap smears. PMID- 15790435 TI - Expression of copper-transporting P-type adenosine triphosphatase (ATP7B) as a prognostic factor in human endometrial carcinoma. AB - OBJECTIVE: Copper-transporting P-type adenosine triphosphate (ATP7B) has been reported to be associated with cisplatin resistance in vitro. However, the clinical significance of this transporter has not previously been addressed in endometrial carcinoma. Our goal was to investigate if ATP7B is expressed in endometrial carcinoma and whether its expression correlates with prognosis. METHODS: We performed immunohistochemical analysis of ATP7B using a monoclonal antibody against ATP7B in 51 endometrial endometrioid adenocarcinomas. 27 of 51 patients were treated with cisplatin-based chemotherapy after surgery. RESULTS: Cytoplasmic staining of tumor cells was observed in 37.3% (19/51 cases) of the analyzed carcinomas and no staining was observed in adjacent non-neoplastic cells. ATP7B positivity in the degree of differentiation of G2 and G3 carcinoma was significantly higher than that of G1 carcinoma (P = 0.019). The patients with ATP7B-positive tumors had a worse prognosis than that with ATP7B-negative tumors in overall survival and disease-free survival, respectively (P < 0.01). CONCLUSIONS: These findings suggest that overexpression of ATP7B expression in endometrial carcinoma is correlated with unfavorable clinical outcome in patients treated with cisplatin-based chemotherapy. ATP7B expression could be considered as a prognostic factor in patients with endometrial carcinoma. PMID- 15790437 TI - Increased expressions of claudin-1 and claudin-7 during the progression of cervical neoplasia. AB - OBJECTIVES: Claudin proteins represent a large family of integral membrane proteins crucial for tight junction formation and function and they have been shown to be expressed differently in various cancers. We investigated whether the expressions of claudin-1 and claudin-7 are associated with the progression of uterine cervical neoplasia. METHODS: We analyzed 89 formalin-fixed paraffin embedded cervical tissues that included 10 normal cervical epithelium, 19 low grade squamous intraepithelial lesion (LSIL), 20 high-grade squamous intraepithelial lesion (HSIL), 20 invasive squamous cell carcinoma (ISCC) without lymph node (LN) metastasis, and 20 ISCC with LN metastasis. The expressions of claudin-1 and claudin-7 were determined by immunohistochemistry. RESULTS: The expressions of claudin-1 and claudin-7 were undetectable in normal cervical squamous epithelium, but had variable staining in the basal layer of normal endocervical glands. The expressions of both proteins, mainly as membranous staining, gradually increased in accordance with the progression from LSIL to HSIL and ISCC (both P values are <0.001) and were detected in all cases of ISCC. CONCLUSIONS: These results suggest that claudin-1 and claudin-7 may play a significant role in tumor progression of cervical neoplasia and may represent useful markers for malignant transformation of cervical squamous cells. Further studies would likely result in the development of novel approaches for early detection and therapy for this disease. PMID- 15790436 TI - p53 and mdm2 as prognostic indicators in patients with epithelial ovarian cancer: a multivariate analysis. AB - OBJECTIVE: The aim of the study was to investigate the prognostic significance of p53 and mdm2 protein expressions in epithelial ovarian cancer and their relationship with the clinicopathological variables. METHODS: Tumor biopsy specimens from 82 patients who were homogenously treated were examined immunohistochemically for expression of p53 and mdm2 proteins. Univariate and multivariate analyses were performed for prognostic factors, and correlations with clinicopathological parameters were examined. RESULTS: Fifty-four percent and 33% of cases stained positive for p53 and mdm2, respectively. p53 expression was associated with serous type, higher grade, positive cytology, residual tumor and stage of the disease. mdm2 expression predicted of chemosensitivity and it was related with higher grade but not with other clinicopathological variables. Significantly poorer survival was seen for those with p53 (P < 0.05) or mdm2 (P < 0.01) positive tumors than those with negative p53 or mdm2 staining. Coexpression of p53 and mdm2 was also related to poor outcome (P < 0.05). Multivariate analysis revealed that FIGO stage, mdm2 expression, response to chemotherapy and optimal cytoreduction were significant independent prognostic and predictive factors of survival. CONCLUSION: Although our findings showed that mdm2 may be used as a prognostic indicator in patients with epithelial ovarian cancer, these results should be supported by more and larger studies. PMID- 15790438 TI - A randomized phase II study of paclitaxel with carboplatin +/- amifostine as first line treatment in advanced ovarian carcinoma. AB - OBJECTIVE: Will amifostine (A) protect against chemotherapy-induced neuro- and myelotoxicity. PATIENTS AND METHODS: Ninety ovarian cancer patients were randomized to receive standard paclitaxel + carboplatin without (PC) or preceded by amifostine 740 mg/m(2) (PC + A). RESULTS: The mean baseline values of hemoglobin, leukocyte, and platelets were slightly lower in the amifostine group, but the mean percentual decrease of these parameters after each treatment cycle showed no difference between both arms. Symptoms of neurotoxicity remained absent in 40% PC vs. 49% PC + A cycles; sensory neurotoxicity grade I occurred in 45% vs. 48% and grade II in 12% PC vs. 2% of PC + A cycles (overall P < 0.001). Nausea grade II was reported in 2% vs. 6% (P = 0.007) and vomiting grade II in 1% of PC vs. 8% PC + A cycles (P < 0.001). Amifostine was temporarily interrupted in five patients due to hypotension, but no dose reductions were indicated. Quality of life questionnaires showed no difference in neurotoxicity scores between both study arms at treatment completion. The median progression-free survival was 16 vs. 22 months (n.s.) for PC and PC + A patients. In a pooled analysis of four randomized studies, amifostine diminished the risk of developing neurotoxicity grade II-III (Odds Ratio 0.3, 95% confidence interval 0.15-0.63, P < 0.05), but had no effect on the risk for bone marrow toxicity. CONCLUSION: Amifostine shows only minor but significant activity in diminishing neurotoxicity without preventing paclitaxel + carboplatin-induced bone marrow toxicity. PMID- 15790439 TI - Molecular cytogenetics of ovarian granulosa cell tumors by comparative genomic hybridization. AB - OBJECTIVE: Patients with stage I granulosa cell tumors (GCTs) may occasionally develop metastasis, which is hard to predict using pathologic criteria. It is interesting to elucidate whether certain chromosomal imbalances (CIs), detected by comparative genomic hybridization (CGH), could be useful prognostic markers. METHODS: CGH was used to identify CI(s) in 37 adult-type GCTs from 36 women. Nonrandom CIs were compared with clinical and pathological features to evaluate their significance as a prognostic marker. RESULTS: Twenty-two (61%) of the 36 primary tumors had CIs. One woman's tumor showed identical CIs to another tumor that occurred in contralateral ovary 2 years later, supporting a metastatic nature. The nonrandom CIs included losses of 22q (31%), 1p33-p36 (6%), 16p13.1 (6%), and 16q (6%) and gains of 14 (25%), 12 (14%), and 7p15-p21 (6%). No tumor exhibited high-level amplification. The associations between each CI and pathological features, including the growth pattern, tumor size, and mitotic activity, were not evident. The only CI repeatedly detected in tumors with metastasis was monosomy 22, which presented in 2 of the 4 cases with metastasis but also in 2 of the 5 cases without recurrence for more than 5 years. CONCLUSIONS: Monosomy 22 was the most common CI in GCTs, which often coexisted with trisomy 14 (in 55% cases). Deletion of 22q seems to be, albeit not very specific, associated with the risk of early metastases of stage I disease. The role of loss-of-function mutation(s) of certain putative tumor suppressor gene(s) on 22q is worthy of further investigations. PMID- 15790440 TI - The role of secondary cytoreductive surgery for recurrent ovarian cancer. AB - OBJECTIVE: The aim of this study was to assess the survival benefit of salvage surgical cytoreduction in patients with recurrent ovarian cancer and compare the surgical outcome with salvage chemotherapy alone. METHODS: Seventy-five patients with recurrent ovarian cancer were reviewed for possible benefits of salvage therapy. Forty-four had salvage surgery and 31 patients had salvage chemotherapy alone for the treatment of gross recurrent disease. All patients had been clinically free of disease more than 6 months from the completion of primary treatment. RESULTS: A macroscopically complete surgical cytoreduction was obtained in 34 (77%) patients. Survival was significantly longer in patients who had salvage surgery compared to those who had salvage chemotherapy alone (P = 0.03). Moreover, survival was significantly longer in patients who were completely cytoreduced compared to those who were not completely cytoreduced and those who were not operated (P = 0.007 and P = 0.005, respectively). CONCLUSIONS: Macroscopically complete surgical cytoreduction significantly improves further survival of recurrent ovarian cancer patients. However, we remain in need to evaluate the debulkability of tumor before surgery to maximize the survival benefit and minimize the number of ineffective surgeries. PMID- 15790441 TI - Histologic subtypes and laterality of primary epithelial ovarian tumors. AB - OBJECTIVES: To determine if the likelihood of bilateral primary ovarian tumors differs by histologic subtype. METHODS: Using data collected by the Surveillance Epidemiology and End Results (SEER) program, the analysis included 22,328 women 25-84 years of age who were diagnosed with a borderline or malignant epithelial ovarian tumor during 1992-2000, categorized as to laterality and histologic subtype. RESULTS: Malignant serous tumors were bilateral in 57.5% of cases. Corresponding figures for mucinous, clear cell, endometrioid and other epithelial tumors were 21.3%, 13.3%, 26.8%, and 35.6%, respectively. Borderline serous tumors were bilateral in 29.8% of the cases compared to only 7.0% of mucinous tumors. The tendency for serous tumors to present as bilateral was consistent across all categories of race, age, and stage. CONCLUSIONS: Serous tumors of the ovary are more commonly bilateral than ovarian tumors of other histologic subtypes. The reasons for this tendency remain to be determined. PMID- 15790442 TI - Results after laparoscopic management of serous borderline tumor of the ovary with peritoneal implants. AB - OBJECTIVE: The aim of this study is to assess the clinical outcomes of laparoscopic treatment of borderline ovarian tumor (BOT) with peritoneal implants. METHODS: Retrospective analysis of patients treated initially and/or for recurrent disease using a laparoscopic approach for a stage II or stage III BOT between January 2001 and January 2004. RESULTS: Nine patients underwent a laparoscopic pure treatment of stage II/III serous borderline tumor. Three of them had a previous history of BOT. Three patients had a stage II and 6 a stage III disease. A conservative management was performed in 7 patients. Laparoscopic treatment of peritoneal implants included: omentectomy (or omental biopsies) in 4 patients and/or large peritoneal resection in 5 patients (pelvic peritoneum in all patients associated with peritonectomies of paracolic gutters in 2 and of the peritoneum of the right diaphragmatic peritoneum in 3). Implants were nonivasive in 8 patients. Each of implant had a size <5 mm. Four patients recurred, 3 of them had a borderline ovarian recurrence after conservative management. Two patients had peritoneal disease found during a second-look surgery (associated with ovarian recurrence in 1). Three spontaneous pregnancies were observed. All patients are alive without evidence of disease with a median time of follow-up of 35 months following the laparoscopic treatment. CONCLUSION: Our series suggests that laparoscopic treatment of patients with BOT associated with small size non invasive implants is feasible and seem to be safe. The main indication of this management consists in young patients treated conservatively to preserve their fertility. PMID- 15790443 TI - Gynecologic cancer treatment and the impact of cancer-related infertility. AB - OBJECTIVE: The objective of this study was to empirically assess the emotional and sexual impact of cancer-related infertility in women with a history of gynecologic cancer. METHOD: Women with a history of gynecologic cancer were approached during their gynecologic oncology clinic appointment; they were provided a description of the study and asked to participate. All participants completed a one-time self-report survey. We present data acquired via the following methods: Center for Epidemiologic Studies-Depression Scale (CES-D), Impact of Events Scale (IES), Modified Inventory of Traumatic Grief (M-ITG), Female Sexual Function Index (FSFI), and the Menopausal Symptom Checklist. RESULTS: The study sample consisted of 20 women, ages 27 to 49 years (mean, approximately 40 years), who had undergone treatment for cervical (40%), ovarian (20%), or uterine (40%) cancer. Forty percent of the sample reported depressive symptoms as measured by the CES-D, with 35% of the women experiencing moderate to severe levels of distress as measured by the IES. The women in this sample experienced dissatisfaction with their overall sex lives (67%), pain during vaginal penetration (62%), and low levels of sexual desire (56%). CONCLUSIONS: The preliminary findings of this study indicated that feelings of depression, grief, stress, and sexual dysfunction are being experienced by women with a history of gynecologic cancer who have lost their fertility as a result of their cancer treatment. PMID- 15790444 TI - The expression of cyclooxygenase-2, VEGF and PGs in CIN and cervical carcinoma. AB - OBJECTIVE: To study the expression of cyclooxygenase-2 (COX-2), vascular endothelial growth factor (VEGF) and prostaglandins (PGs) in cervical tissues of differential pathological types, especially in cervical intraepithelial neoplasia (CIN) and cervical carcinoma, and their possible relationships in carcinogenesis. METHODS: Tissue blocks and blood samples from 20 normal cervix women, 20 cervix inflammation patients, 20 CIN patients and 40 patients with cervical carcinoma, respectively, at our institutions from February 2000 to March 2002 were available for this study. COX-2 mRNA was examined by reverse transcription-polymerase chain reaction (RT-PCR). COX-2 and VEGF proteins were measured by immunohistochemistry using monoclonal antibodies to them. PGs (PGE2, TXB2, 6-k-PGF(1alpha)) were detected by radioimmunoassay (RIA). RESULT: The overall positive expression of COX-2 and the quantity of PGs, especially PGE2 in inflammation, CIN and cervical carcinoma was higher and much higher than that in normal cervix (P < 0.001), There was a close relationship between COX-2 and PGs. The positive expression rate of VEGF in cervical carcinoma was higher than that in normal, inflammatory and CIN cervix, respectively (P < 0.001). VEGF protein was occasionally expressed in CIN cervix (15%). There was no association among COX-2, VEGF and clinicopathological parameters in cervical carcinoma. The expression of COX-2 and VEGF in cases with tumor in diameter more than 4 cm (90.9%, 72.7%) was higher than those with smaller tumor (86.2%, 51.7%). CONCLUSIONS: The COX-2 probably is a gene involved early in carcinogenesis of cervical carcinoma by increase of PGs, and accelerates the progress of tumor by increase of PGs and VEGF. Therefore testing the expression of PGs may be a prognosis marker for clinical diagnosis. PMID- 15790445 TI - The influence of age and co-morbidity on treatment and prognosis of ovarian cancer: a population-based study. AB - OBJECTIVES: With the rising mean age, more patients will have one or more other serious diseases at the time of diagnosis of ovarian cancer (co-morbidity). In this study, the independent effects of age and co-morbidity on the application of treatment guidelines and prognosis were evaluated. METHODS: All patients with epithelial ovarian cancer diagnosed between 1995 and 2001 in the southern part of The Netherlands (N = 1116) were included. RESULTS: The prevalence of co-morbidity increased from 34% of the age group <70 to 63% of the older age group. Eighty three percent of the patients with FIGO stage II or stage III younger than 70 years underwent the advised treatment (combination of surgery and chemotherapy) compared to only 45% of the patients aged 70 or older. In a multivariable analysis age, FIGO stage, presence of co-morbidity, and year of diagnosis seemed to be independent predictors of receiving the advised treatment. In multivariable analyses age 70 + (HR = 1.3, 95% CI = 1.03-1.7) and the use of both surgery and chemotherapy (HR = 0.4, 95% CI = 0.3-0.6, reference is only surgery) were independent prognostic factors for overall survival. CONCLUSIONS: Even in the absence of co-morbidity, standard combination therapy was prescribed significantly less often for elderly patients with FIGO II or III ovarian cancer. Age and combined treatment of surgery and platinum-based chemotherapy were independent prognostic factors. Co-morbidity did not seem to have a prognostic effect. PMID- 15790446 TI - Overexpression of dihydrodiol dehydrogenase is associated with cisplatin-based chemotherapy resistance in ovarian cancer patients. AB - OBJECTIVE: Results of a recent study on human ovarian cancer cell lines indicated that overexpression of dihydrodiol dehydrogenase (DDH) was associated with resistance to cisplatin and disease progression. We examined the relationships between DDH expression and chemotherapy resistance in ovarian cancer patients. METHODS: Using immunohistochemistry, expression of DDH was measured in 41 patients with epithelial ovarian cancers. All patients underwent primary debulking surgery, followed with six cycles of cisplatin-based chemotherapy. Normal ovarian tissues were obtained from patients with benign gynecologic diseases (n = 14). Expression of DDH was confirmed by reverse transcription polymerase chain reaction. The correlation between DDH expression and clinico pathological parameters was analyzed by statistical analysis. Difference of progression-free survivals between different groups was compared by a log-rank test. RESULTS: Eighteen ovarian cancer samples (43.9%) expressed DDH at a moderate to strong level. This marked a significant difference from the negligible expression (1/14, 7.1%) found in the control group (P = 0.02). Of interest, the clear cell adenocarcinoma revealed DDH overexpression (75%) and mucinous adenocarcinoma revealed low DDH expression (16.7%), although DDH expression did not show any significant variation according to different histotypes. DDH overexpression was found in a statistically significantly higher percentage of cisplatin-resistant cases (n = 8/11; 72.7%) than in cisplatin sensitive cases (n = 9/27; 33.3%) (P = 0.037). Using multivariate analysis, only DDH retained as an independent role in predicting a poor chance of response to cisplatin-based treatment. DDH overexpression cases (median 12 months, 95% confidence interval 4-20) demonstrated a shorter progression-free survival than DDH-negative cases (median 28 months, 95% confidence interval 23-33), but this result did not reach the statistical significance (P = 0.1742). In the advance stage, the DDH-positive group has a shorter PFS as compared with DDH-negative group, and this result closely approaches the statistical significance (P = 0.0669). CONCLUSIONS: DDH is expressed in a high percentage of primary ovarian tumors and its expression may be associated with cisplatin-based chemotherapy resistance. The possible prognostic role of DDH in ovarian carcinoma deserves further study. PMID- 15790447 TI - Acute diarrhea during pelvic irradiation: is small-bowel volume effect different in gynecologic patients with prior abdomen operation or not? AB - OBJECTIVE: To evaluate volume effect of small bowel for diarrhea during pelvic irradiation in gynecologic patients with or without prior abdomen operation. METHODS: From January 1996 through December 2003, 759 patients undergoing 4-field pelvic irradiation for cervical or uterine cancer were analyzed. Whole pelvic (WP), modified whole pelvic (MWP), or lower pelvic (LP) irradiation were delivered initially. According to contrast medium within small bowel in simulation films, we categorized the small-bowel volume of full dose related to WP fields as small-volume and large-volume groups. We recorded the severity of diarrhea until 39.6 Gy/22 fractions of pelvic irradiation. The actuarial rates of overall and moderate to severe diarrhea were compared among different groups. RESULTS: Significantly more large-volume distribution (85%) was noted in patients >60 years without prior operation (P < 0.001). Large-volume distribution was 53%, 65%, and 82% in post-operative patients with no diarrhea, mild diarrhea, and moderate to severe diarrhea (P = 0.002), respectively. The corresponding rate was 79%, 77%, and 80% in patients without prior abdomen operation (P = 0.869). In multivariate analysis, prior operation with LP fields (P = 0.005) and prior operation with small volume (P = 0.031) were significantly protective factors for overall diarrhea. The latter was also a protective factor for moderate to severe diarrhea (P = 0.026). Prior operation could diminish overall diarrhea in patients without simultaneous large-field (WP or MWP) and large-volume. Large volume was a significant factor of overall (P = 0.014) and moderate to severe (P = 0.004) diarrhea in large-field patients with operation. The volume effect did not exist in those patients without operation. CONCLUSION: Age and operation can change small-bowel distribution. Prior operation may attenuate diarrhea if irradiated volume of small bowel is small. There is a volume effect in post-operative rather than non-operative patients receiving large-field irradiation. More practical dose-volume evaluation of small bowel may be applied for volume effect in gynecologic patients without prior operation. PMID- 15790449 TI - Lymph node spread in stage III or IV primary peritoneal serous papillary carcinoma. AB - BACKGROUND: The aim of this study was to determine the rates and topography of pelvic and para-aortic nodal involvement in patients with stage III or IV primary peritoneal serous papillary carcinoma (PSPC). METHODS: Retrospective review of 19 women who underwent a systematic bilateral pelvic and para-aortic lymphadenectomy. RESULTS: The overall frequency of lymph node involvement was 63% (12/19). Eighteen patients underwent complete resection of peritoneal disease. Only 4 patients underwent this procedure as part of their initial surgery (before chemotherapy). The frequency of pelvic and para-aortic metastases was 58% (11/19) and 58% (11/19), respectively. When para-aortic nodes were involved, the left para-aortic chain above the level of the inferior mesenteric artery was the site most frequently involved (72%). The event-free survival of the 18 patients without macroscopic disease at the end of debulking surgery was significantly correlated with the nodal status. None of the patients with positive nodes developed recurrent disease in abdominal nodes. CONCLUSIONS: The rate of nodal involvement in patients with PSPC is high. The topography of nodal spread is similar to that of ovarian cancer. Lymphadenectomy has a prognostic value. PMID- 15790448 TI - Extended-field radiotherapy and high-dose-rate brachytherapy with concurrent and adjuvant cisplatin-based chemotherapy for locally advanced cervical cancer: a phase I/II study. AB - OBJECTIVE: The purpose of this prospective study was to investigate the toxicity and efficacy of integrating extended-field para-aortic and pelvic external radiation, high-dose-rate intracavity brachytherapy, and concurrent and adjuvant cisplatin-based chemotherapy for locally advanced cervical cancer. METHOD: A phase I/II study was performed from 1998 to 2003 including sixty-three patients with both clinical FIGO and MRI/CT-based TNM stage IIB-IVA cervical cancer. Patients were treated with extended-field external radiation to the para-aortic and pelvic regions with 45 Gy in 25 fractions, and an additional boost to the gross nodes to 50.4 Gy and the parametrium to 59.4 Gy. Patients also received a high-dose-rate (HDR) intracavity brachytherapy with doses of 22-31 Gy to point A in 4-6 fractions. Concurrently, two cycles of cisplatin (50-80 mg/m(2)) were administered in weeks 1 and 5 during radiotherapy, as well as two cycles of cisplatin (60-80 mg/m(2)) for 1 day and 5-fluorouracil (600-800 mg/m(2)) for 4 days at 1 and 2 months after completion of radiotherapy. The treatment-related acute and late side effects were evaluated using RTOG criteria, and the disease control and survival rate were calculated using the Kaplan-Meier method. The median follow-up interval was 36 months. RESULTS: All sixty-three patients completed the planned extended-field radiotherapy and high-dose-rate brachytherapy with 2 concurrent cycles of cisplatin. Fifty-eight (92%) patients received 2 cycles of the post-radiation adjuvant chemotherapy of cisplatin and 5 fluorouracil. RTOG grade III acute toxicity was gastrointestinal (2%) and hematological (10%). No patient had grade IV acute toxicity. Late grades III-IV morbidity actuarial risk of 6% at 6.5 years primarily involved the injuries to the bowels requiring surgical intervention for intestinal obstruction or fistula formation. Initial sites of recurrence were locoregional failure alone (pelvic and para-aortic regions within the radiation field), 3%; distant metastases only, 8%; and locoregional failure plus distant metastases, 8%. The observed rates at 3 year and 5-year of locoregional control, freedom from distant metastasis, and overall survival were 86% and 86%, 81% and 81%, and 81% and 77%, respectively. CONCLUSION: Incorporating HDR brachytherapy into a regimen including concurrent chemotherapy and extended radiation appears safe and effective. PMID- 15790450 TI - Sensitization of cervical cancer cell lines to low-dose radiation by retinoic acid does not require functional p53. AB - OBJECTIVE: Current therapy for cervical cancer includes radiation therapy. Retinoic acid (RA) can increase the sensitivity of cervical cancer cell lines to radiation. The mechanism of this sensitization may not involve the p53 protein because the human papillomavirus (HPV) E6 protein, which is present in the majority of cervical cancers, promotes p53 degradation. The objective of this study was to determine if p53 is involved in the mechanism of RA radiosensitization. METHOD: The effects of radiation on cervical (SiHa, CC-1, and C33a) and vulvar (SW962) cancer cell lines under various experimental conditions were evaluated using clonogenic, Coulter Counter, electrophoretic mobility shift (EMSA) and a multi-probe RNase protection assay of p53-inducible genes. RESULTS: RA (5 microM 9-cis-RA) radiosensitized the SiHa and CC-1 cell lines that contain HPV-degraded p53, but did not radiosensitize the SW962 cell line, which is HPV negative and contains wild-type p53, nor the C33a cell line, which contains mutant p53 (R273C). Expression of mutant p53 (R273H) in SiHa cells increased the growth rate, but did not prevent RA-induced differentiation or radiosensitization at clinically relevant doses. Inhibition of p53 transactivation with pifithirin alpha did not prevent RA radiosensitization of SiHa at 5 Gy. RA repressed c-fos mRNA expression in control and irradiated SiHa cultures, but did not repress bcl x(L), p53, GADD45, p21, bax, bcl-2, or mcl-1 mRNA expression. CONCLUSIONS: The mechanism of RA radiosensitization does not require functional p53 and may involve c-fos in cervical cancer cell lines. PMID- 15790451 TI - Sentinel node localization should be interpreted with caution in midline vulvar cancer. AB - OBJECTIVE: Carcinomas of the vulva situated on the midline or close to it, are supposed to have a bilateral lymphatic drainage. The aim of this study was to evaluate sentinel node identification in these tumors. METHODS: Between April 2002 and February 2004, 17 patients with operable vulvar cancer situated on, or close to the midline were entered in a prospective study. All patients underwent sentinel node identification with (99m)Tc-labelled nanocolloid (preoperative lymphoscintigraphy and intraoperative use of a handheld probe). Depending on the surgeon, intraoperative blue dye was associated. Radical excision of the tumor and routine bilateral lymphadenectomy were then performed. Sentinel nodes were sent separately for histologic examination. Negative sentinel nodes on hematoxylin/eosine were further examined with immunohistochemistry. RESULTS: One or more sentinel nodes were identified in the 17 patients and in 21 of the 34 groins. In 5 patients, the sentinel nodes were metastatic. There was no false negative (negative sentinel node and metastatic non-sentinel node). In 13 patients, lymphoscintigraphy and then intraoperative identification suggested a unilateral drainage of the tumor with sentinel nodes localized in only one groin. Among these 13 patients, 3 groins with no sentinel node identified contained in fact massively metastatic nodes. CONCLUSION: Unilateral finding of a sentinel node in tumors of the midline does not preclude a metastatic node in the other groin. Lymph node assessment should remain bilateral in these lesions. PMID- 15790452 TI - Snail and Slug are major determinants of ovarian cancer invasiveness at the transcription level. AB - OBJECTIVES: The transcriptional factors Snail and Slug have been reported to be important in cell migration during development and also during tumor metastasis. Their expression and role in ovarian cancer, hitherto unexplored, was examined to understand the molecular events in ovarian cancer metastases since the latter is responsible for the high degree of mortality associated with the disease. METHODS: Ectopic expression of mSnail and mSlug in the epithelial ovarian cancer cell line SKOV3 was carried out and stable clones were selected. These were used to examine specific repression of the adherens, tight and desmosomal junction components by the two transcription factors. Furthermore, functional implications with respect to enhanced migration of cells, tumorigenecity and metastasis were also studied. RESULTS: The ectopic expression of Snail or Slug resulted in epithelial-mesenchymal transition (EMT), enhanced motility, invasiveness and tumorigenecity in the cell line SKOV3. In addressing the mechanism by which Snail and Slug lead to loss of intercellular adhesion, specific repression of adherens junction components (E-cadherin and betacatenin), tight junction components (Occludin and ZO-1) and desmosomal junction components (Dsg2) were observed. Snail suppresses expression of adherens and tight junction components, while Slug suppresses expression of all the three junction components; concertedly, bringing down the intercellular adhesion between cells. Further activation of these transcriptional factors in hypoxic conditions revealed a rapid upregulation of Slug expression as an immediate reaction that probably triggers off a signaling cascade leading to Snail expression. CONCLUSIONS: These results indicate distinct roles of the transcriptional factors Snail and Slug during ovarian cancer metastasis and cell survival through mediation of EMT. PMID- 15790453 TI - A comparison between different postoperative treatment modalities of uterine carcinosarcoma. AB - OBJECTIVE: Uterine carcinosarcomas are highly aggressive neoplasms with no established effective adjuvant therapy. The aim of the present study was to compare between the outcome in three medical institutions in each of which a different postoperative treatment modality was preferred, namely, chemotherapy in one, whole pelvic irradiation (WPI) in another, and sequential treatment (i.e., chemotherapy followed by WPI) in the third. METHODS: The hospital records of all 49 uterine carcinosarcoma patients diagnosed and operated from 1995 to 2003 in the three institutions were reviewed. Non-parametric test was used to compare the median age between the treatment groups. Survival was calculated using the Kaplan Meier method and compared by the log-rank test. Cox proportional hazard regression model was used to assess the effect of treatment type on survival after adjustment for stage. RESULTS: Only about half of the patients (51%) had stage I at diagnosis and the majority of the patients (83.7%) had postoperative adjuvant treatment. The overall 5-year survival of the 41 patients that had postoperative treatment was 49.6%. The highest median survival and 5-year survival rate was observed in the sequential treatment group. Controlling for stage, this treatment modality was associated with a significant decrease in mortality of about 80% when compared to postoperative chemotherapy alone, and a non-significant decrease in mortality of about 50% when compared to WPI alone (HR = 0.20; 95% CI 0.04-0.99, P = 0.049 and HR = 0.50; 95% CI 0.1-2.32, P = 0.4, respectively). CONCLUSIONS: The improved outcome in patients who received postoperative sequential treatment seems to indicate that further exploration of this treatment modality is justified. PMID- 15790454 TI - Adjuvant sequential chemotherapy and radiotherapy in uterine papillary serous carcinoma. AB - PURPOSE: To evaluate the efficacy and toxicity of adjuvant combination of sequential chemotherapy followed by radiotherapy in uterine papillary serous carcinoma (UPSC). METHODS AND MATERIALS: From April 1994 to June 2003, 26 patients (median age 61.7 years, range 46.9-78.4) with UPSC were treated with a platinum-based chemoradiation protocol after definitive surgery. 9 patients were assigned as stage I (35%), 4 were stage II (15%), 11 were stage III (42%), and 2 were stage IV (8%) according to the FIGO staging for gynecological cancers. All patients underwent total hysterectomy, salpingo-oophorectomy, pelvic +/- perioartic lymph nodes dissection/sampling, omentectomy, and peritoneal washing. The adjuvant chemoradiation protocol consists of 4 cycles of platinum-based chemotherapy followed by pelvic irradiation and vaginal vault brachytherapy. In selected stage I patients with no or minimal myometrial invasion, only vault brachytherapy was given after adjuvant chemotherapy. RESULTS: After a median follow-up of 28 months (range 9-113 months), 14 (54%) patients were alive and free of disease. 12 out of these 14 patients were FIGO stage I/II. 9 patients (35%) had died (8 from distant metastases). The Kaplan-Meier 2-year and 5-year survival estimates were 69.5% and 57%, respectively. Only 4 (15%) patients had pelvic recurrence. None of the patients developed local vault recurrence. The treatment was well tolerated, only 1 patient developed congestive cardiac failure from the chemotherapy and 6 patients had grade 2 peripheral neuropathy on follow up. CONCLUSION: In our series of UPSC patients treated with adjuvant chemotherapy followed by radiotherapy, local control can be achieved in a majority of patients. Distant failure remains the major cause of mortality. Further investigations into finding a more effective systemic therapy are required if improvement in outcome for this form of uterine cancer is to be achieved. PMID- 15790455 TI - Plasma kinetics and uptake by the tumor of a cholesterol-rich microemulsion (LDE) associated to etoposide oleate in patients with ovarian carcinoma. AB - OBJECTIVES: Previously, we reported that etoposide oleate associated to a cholesterol-rich microemulsion (LDE) is taken up by malignant cells overexpressing low-density lipoprotein (LDL) receptors. The association is stable, preserves antiproliferative activity of the drug, and reduces toxicity to animals. Here, we determined in patients the plasma kinetics of LDE-etoposide oleate and verified whether the complex concentrates in ovarian carcinomas. METHODS: [(3)H]-etoposide oleate associated to LDE labeled with [(14)C] cholesteryl oleate was intravenously injected into four ovarian carcinoma patients (50 +/- 8.7 years) 24 h before surgery. Blood samples were collected over a 24-h period to determine the radioactivity plasma decay curves, and the plasma fractional clearance rate (FCR) was calculated by compartmental analysis. Specimens of tumors and normal ovaries excised during the surgery were collected for lipid extraction and radioactive counting. RESULTS: FCRs of LDE label and of the drug were similar (0.0985 and 0.1722, respectively, P = 0.2422). [(14)C]-LDE uptake was 4.9 times and [(3)H]-etoposide oleate uptake was 4.1 times greater in the ovarian tumors than in the contralateral normal ovaries (LDE uptake, in cpm/g = 560 +/- 171 and 146 +/- 59; etoposide oleate uptake = 346 +/- 75 and 103 +/- 56, respectively). CONCLUSIONS: Most of the drug is retained in the microemulsion particles until its removal from the circulation and internalization by the cells. In addition, LDE-etoposide oleate has the ability to concentrate in malignant ovarian tissues. Therefore, the complex may be used to direct and concentrate etoposide oleate in ovarian carcinomas. PMID- 15790456 TI - FDG-PET for management of cervical and ovarian cancer. AB - OBJECTIVE: To assess the diagnostic performance of Positron Emission Tomography using fluorodeoxyglucose (FDG-PET) in comparison to conventional imaging modalities in the assessment of patients with cervical and ovarian cancer. METHODS: Studies published between 1966 and 2003 were identified using an OVID search of the MEDLINE database. Inclusion criteria were use of a dedicated scanner, resolution specified, >/=12 human subjects, clinical follow-up >/=6 months or histopathology as reference standard, and sufficient data provided to construct a two-by-two table. Two reviewers independently abstracted data regarding sensitivity and specificity of PET. RESULTS: 25 studies (15 cervical cancer, 10 ovarian cancer) met inclusion criteria for full text review. For cervical cancer, pooled sensitivity and specificity of PET for aortic node metastasis are 0.84 (95% CI 0.68-0.94) and 0.95 (0.89-0.98). Pooled sensitivity and specificity for detection of pelvic node metastasis are: PET, 0.79 (0.65 0.90) and 0.99 (0.96-0.99); MRI, 0.72 (0.53-0.87) and 0.96 (0.92-0.98). Pooled sensitivity for CT is 0.47 (0.21-0.73) (pooled specificity not available). Pooled sensitivity and specificity of PET for recurrent cervical cancer with clinical suspicion are 0.96 (0.87-0.99) and 0.81 (0.58-0.94). For ovarian cancer, pooled sensitivity and specificity to detect recurrence with clinical suspicion are: PET, 0.90 (0.82-0.95) and 0.86 (0.67-0.96); conventional imaging, 0.68 (0.49 0.83) and 0.58 (0.33-0.80); CA-125, 0.81 (0.62-0.92) and 0.83 (0.58-0.96). When conventional imaging and CA-125 are negative, pooled sensitivity and specificity of PET are 0.54 (0.39-0.69) and 0.73 (0.56-0.87), respectively. When CA-125 is rising and conventional imaging is negative, the pooled sensitivity and specificity of PET are 0.96 (0.88-0.99) and 0.80 (0.44-0.97). CONCLUSIONS: There is good evidence that PET is useful for the pre-treatment detection of retroperitoneal nodal metastasis in cervical cancer. There is fair evidence that PET is useful for the detection of recurrent cervical cancer. PET is less useful for the detection of microscopic residual ovarian cancer but has fair sensitivity to detect recurrence in the setting of a rising CA-125 and negative conventional imaging studies. Available studies are limited by low numbers of patients and wide confidence intervals. PMID- 15790457 TI - Vulvar basal cell carcinoma: retrospective study and review of literature. AB - OBJECTIVES: Basal Cell Carcinoma (BCC) is the most common human malignant neoplasm. It does not occur only in anatomical areas commonly exposed to sunlight, but also in relatively protected body sites such as the axillae, the groin and buttocks, and in regions which are entirely protected from the sun, like the vulva. The aim of this study is to assess the impact and clinical features of BCCs in the vulva versus other anatomical sites. METHODS: For this purpose, we retrospectively reviewed all the histological diagnoses of BCCs made between 1995 and 2003 in the Department of Human Pathology and Oncology of the University of Florence, Italy. RESULTS: Out of 3604 cases of BCC, 63 were in the vulva. The average age of patients suffering from the latter was 70.35 years. The average size of the vulvar BCCs was 2.1 cm and 18 (28%) were ulcerated at presentation. CONCLUSIONS: Our study shows that vulvar BCC is not a particularly rare occurrence; in fact it should be suspected whenever lesions that we believe to be inflammatory do not respond to usual treatment. In our experience, mucocutaneous examination of this anatomical area should be performed especially in patients with a history of skin cancer. In case of suspicion, a biopsy is recommended to obtain a preoperative confirmatory diagnosis. PMID- 15790458 TI - Survival benefit for patients with advanced-stage transitional cell carcinomas vs. other subtypes of ovarian carcinoma after chemotherapy with platinum and paclitaxel. AB - OBJECTIVE: Transitional cell carcinoma (TCC) of the ovary is a less well recognized histological type of ovarian carcinoma resembling TCC of the urinary bladder. A better prognosis due to a better chemosensitivity of ovarian TCC has been suggested. It was the aim of the present retrospective study to compare incidence and outcome of patients with TCCs and other subtypes of ovarian carcinoma from a large homogeneous collective of patients with primary advanced stage ovarian carcinoma. METHODS: H and E-stained sections from a total of 302 cases from a prospective randomized, multi-center, phase III study of patients with ovarian cancer, FIGO-stages IIB-IV, comparing cisplatin plus paclitaxel (PT) with paclitaxel plus carboplatin (TC) were available for histological retyping of ovarian carcinomas applying current WHO criteria. Kaplan-Meier survival analysis was performed. RESULTS: 16 of 302 tumors (5.3%) were diagnosed as TCC. Only 1 of the 16 TCCs had been previously diagnosed as such by referring pathologists. TCCs were associated with smaller preoperative extraovarian tumor and with smaller postoperative residual tumor. 5-year survival of patients with TCC was 57% as compared to 31% for patients with ovarian carcinomas of other types (P = 0.03). CONCLUSION: TCC of the ovary seems to be a less well recognized entity. In the current series, TCCs had a significantly better prognosis as compared to all other types of ovarian carcinomas after standardized chemotherapy. A propensity for micronodular rather than macronodular extraovarian spread and better surgical resectability of TCC might contribute to the survival benefit. PMID- 15790459 TI - Paclitaxel, carboplatin, and oral etoposide as initial treatment for advanced ovarian carcinoma: a Minnie Pearl Cancer Research Network phase II trial. AB - PURPOSE: To evaluate the feasibility and toxicity of the combination of paclitaxel, carboplatin, and etoposide in the first-line treatment of patients with stage III or IV adenocarcinoma of the ovary. PATIENTS AND METHODS: Patients entering this trial received paclitaxel 200 mg/m(2), 1 h IV infusion, day 1; carboplatin AUC 6.0 IV, day 1; etoposide 50 mg alternating with 100 mg orally, days 1-10. Patients received 6 courses of treatment, administered at 21-day intervals. All patients were assigned a response category using clinical restaging criteria. The primary endpoints for this trial were progression-free and overall survival. RESULTS: Between October 1996 and April 2001, 52 patients were treated. The overall objective response rate for the 48 evaluable patients was 75%, with 46% complete responses. Amongst the 36 patients with suboptimal disease, median progression-free and overall survivals were 12 and 24 months, respectively. After a median follow-up of 64 months, the median progression-free and overall survival has not been reached for the optimal patients; 5-year progression-free survival is 57% for this group. Treatment-related myelosuppression was common, but myelosuppression-related complications were uncommon, as was grade 3/4 non-hematologic toxicity. No episodes of acute myelogenous leukemia have developed. CONCLUSIONS: The combination of paclitaxel, carboplatin, and oral etoposide was feasible and effective in this patient population. Unlike previous reports, no episodes of acute leukemia were seen following this treatment. Definitive conclusions regarding the benefit of adding etoposide to a taxane/platinum combination will require a comparative trial. PMID- 15790460 TI - Vascular endothelial growth factor (VEGF) up-regulates epidermal growth factor receptor (EGF-R) in cervical cancer in vitro: this action is mediated through HPV E6 in HPV-positive cancers. AB - OBJECTIVES: Epidermal Growth Factor Receptor (EGF-R) up-regulation in cervical cancer cells leads to an increase in cell proliferative Insulin-like Growth Factor II (IGF-II) and Vascular Endothelial Growth Factor (VEGF) and a decrease of the anti-proliferative IGF-binding protein-3 (IGF-BP3). The objectives for this study are: (a) to find if VEGF, in turn, up-regulates EGF-R and down regulates IGF-BP3; (b) to determine if human papilloma virus (HPV-E6) mediates this action of VEGF in HPV-positive cells; and (c) to verify if these effects are reflected in changes in cell proliferation METHODS: We used HPV-positive HeLa (Black), ME-180 and CaSki (Caucasian) and HPV-negative HT-3 (Caucasian) cell lines. (a) Levels of HPV-E6 in the HPV-positive cells were enumerated after treating the cells for 24 h with 20 ng/ml of VEGF using our semi-quantitative immunofluorescent antibody assay. (b) Cellular levels of EGF-R, HPV-E6, IGF-II and IGF-BP3 were enumerated in ME-180 and CaSki cells incubated for 24 h with 5, 10 and 20 ng/ml of VEGF. (c) HPV-negative HT-3 and HPV-positive ME-180 and CaSki cells were incubated with 20 ng/ml VEGF alone or in combination with antibodies to HPV-E6 and EGF-R. HPV-E6 (measured only in HPV-positive cells), EGF-R, IGF-II and IGF-BP3 levels were measured. (d) Cell proliferation was determined using cell proliferation Bradykinine-U colorimetric assay, in HT-3, HeLa and ME-180 cell lines in the presence of VEGF alone and with HPV-E6 antibodies. RESULTS: (a) In all the HPV-positive cell lines, 20 ng/ml VEGF significantly increased (30 50%; P < 0.0001) the HPV-E6. (b) In the ME-180 and CaSki cells, VEGF treatment up regulated EGF-R, IGF-II and HPV-E6 and down-regulated IGF-BP3 in a dose-dependent manner (P < 0.001). (c) These effects of VEGF were eliminated when the HPV positive cells were co-incubated with antibodies to HPV-E6 or EGF-R. In the HPV negative HT-3 cells, VEGF decreased IGF-BP3 while increasing EGF-R and IGF-II levels. Antibodies to EGF-R eliminated these effects (P < 0.0001). (d) Treatment with VEGF resulted in increased cell proliferation in HT-3, HeLa and ME-180 cells; co-incubation with HPV-E6 antibodies abrogated this effect only in the HPV positive cells. CONCLUSIONS: In cervical cancer, VEGF up-regulates EGF-R and down regulates IGF-BP3, thus amplifying the cell proliferative activity of EGF-R. This action of VEGF seems to be mediated, directly through EGF-R or indirectly through HPV-E6 in the HPV-positive cancers, while EGF-R up-regulation appears to play a major role in the HPV-negative cervical cancers. PMID- 15790462 TI - Malignant mixed Mullerian tumor of the fimbriated end of the fallopian tube: origin as an intraepithelial carcinoma. AB - BACKGROUND: A paucity of examples of malignant mixed Mullerian tumors (MMMT) of the fimbriated end of the fallopian tube has been reported. CASE: We report a first case of FIGO Stage IV primary MMMT, heterologous type, in the right fimbria of a 77-year-old woman associated with symptomatic pleural spread who succumbed with recurrent disease 12 months after resection and postoperative paclitaxel and carboplatin chemotherapy. CONCLUSIONS: The identification of intraepithelial carcinoma in this tumor lends support to a role of the epithelial component in fimbrial MMMT histogenesis as seen for MMMT at other anatomic sites. Comparison of the clinical management of these tumors shows prolonged survival of patients whose treatment included postoperative pelvic external radiotherapy. PMID- 15790461 TI - Primary stage I-IIE non-Hodgkin's lymphoma of uterine cervix and upper vagina: evidence for a conservative approach in a study on three patients. AB - BACKGROUND: Non-Hodgkin's Lymphomas (NHL) frequently affect the uterine corpus, cervix, and vagina in cases of advanced disease. However, these organs are rarely the site of origin of this type of neoplasia. Because of the rarity of primary genital tract lymphomas, a standard treatment has not been defined. CASE: Three patients with large B-cell primary Non-Hodgkin's lymphoma of the lower genital tract (vaginal, cervical and cervico-vaginal) presented with bulky lesions and underwent diagnostic evaluation, staging, and chemotherapy with adriamycin containing regimens. All three patients, including two with stage IIE and one with stage IE disease demonstrated complete remission and are alive and well without evidence of disease at 10, 7, and 6 years of follow-up, respectively. CONCLUSIONS: Our observations suggest that young patients with large B-cell lymphomas of lower genital tract stages I-IIE, even with bulky lesions, may benefit from chemotherapy alone as initial treatment. PMID- 15790463 TI - A case of pure-type ovarian squamous cell carcinoma for which combination chemotherapy consisting of paclitaxel and carboplatin was not effective. AB - BACKGROUND: There has been no report on the effectiveness of combination chemotherapy consisting of paclitaxel and carboplatin for pure-type ovarian squamous cell carcinoma (POSCC). CASE: A 56-year-old Japanese woman diagnosed as having pT2c pN1 M0 pure-type ovarian squamous cell carcinoma was treated with combination chemotherapy consisting of paclitaxel and carboplatin every 3 weeks. Four months after the initial chemotherapy, multiple lung tumors appeared and a new tumor in the internal iliac area appeared. She died of disease 12 months after the first treatment. CONCLUSION: Adjuvant chemotherapy consisting of paclitaxel and carboplatin for ovarian squamous cell carcinoma were not effective in this case. PMID- 15790464 TI - Three cases of hemolytic uremic syndrome in ovarian cancer patients treated with combination gemcitabine and pegylated liposomal doxorubicin. AB - BACKGROUND: Hemolytic uremic syndrome (HUS) is a rare coagulation disorder characterized by microangiopathic hemolytic anemia, thrombocytopenia and acute uremia. Reports have described this fatal syndrome in association with cytotoxic agents. To our knowledge, no case reports of HUS in ovarian cancer patients receiving treatment with combination gemcitabine and pegylated liposomal doxorubicin (PLD) have been reported. CASE REPORTS: Three patients with recurrent ovarian carcinoma each developed profound hypertension and peripheral edema while receiving combination gemcitabine and PLD. The first patient had rapid hemolysis, thrombocytopenia, renal failure and respiratory distress. The other patients experienced slowly progressive renal failure and mild hematologic abnormalities. Two of the three patients had favorable outcomes. CONCLUSION: The reported incidence of gemcitabine-induced HUS is rare. Clinicians should suspect HUS if blood pressure elevation or peripheral edema develop. PMID- 15790465 TI - Sigmoid impaction secondary to urinary stones: case report and review of literature. AB - BACKGROUND: This is the first case report of a Miami pouch sigmoid fistula developing passage of urinary stones resulting in the presentation of constipation secondary to impaction. CASE REPORT: A 49-year-old woman who developed a recurrence of invasive squamous cell cervical carcinoma 1 year after pelvis radiation. She then underwent anterior pelvic exenteration and creation of a Miami pouch. Approximately 14 years after the primary radiation therapy and 13 years after the creation of the exenterative procedure, the patient developed a Miami pouch sigmoid fistula. The decision was made at this time to repair the fistula and remove the urinary stones from the sigmoid colon. Postoperatively, the patient remained continent using intermittent catheterization of the pouch and there was no evidence of recurrence of the cancer. CONCLUSION: Conservative management of urinary reservoir complications should always be considered before surgical intervention is attempted. When indicated, surgical management should not be delayed. PMID- 15790466 TI - Yolk sac tumor of the vulva: a case report with long-term disease-free survival. AB - BACKGROUND: Yolk sac tumor (YST) of the vulva is extremely rare. Seven cases of vulvar YST have been reported to the literature. Due to the rarity of tumors, the appropriate choice of treatment may remain unclear. CASE: A 30-year-old woman presented with a 3.5-cm right labial mass. Excisional biopsy showed YST with predominant solid pattern. Three weeks after excision, right inguinal lymph node biopsy revealed metastatic tumor. The serum alpha-fetoprotein (AFP) was not elevated. Cisplatin-based chemotherapy was administered, followed by pelvic and groin irradiation. The patient was free of disease 90 months after the diagnosis. CONCLUSION: Local excision of tumor with adjuvant cisplatin-based chemotherapy can be justified for vulvar YST. Inguinal lymphadenectomy is recommended because metastasis may occur early. Adjuvant radiation therapy may help to control the disease. Tumor size of 5 cm or less may be a favorable prognostic factor. Serum AFP level may not be a sensitive marker for follow-up of vulvar YST. PMID- 15790467 TI - Primary omental gestational choriocarcinoma ascertained by deoxyribonucleic acid polymorphism analysis. AB - BACKGROUND: Abdominal choriocarcinoma is extremely rare. It is important to examine whether the disease is primary or metastatic and gestational or non gestational. CASE: A 26-year-old nulli-gravid woman underwent laparoscopy for presumed ectopic pregnancy. The uterus, ovaries and fallopian tubes surrounded by hemoperitoneum were unremarkable. A hemorrhagic 7-cm-sized tumor was identified on the greater omentum and excised. Histology was consistent with choriocarcinoma. Analysis of human leucocyte antigen (HLA) gene polymorphism on deoxyribonucleic acid (DNA) demonstrated that tumor DNA contained both HLA locus antigens of patient and of her husband. Clinical remission was achieved with six courses of chemotherapy. CONCLUSION: To our knowledge, this is the first reported case of choriocarcinoma that occurred primarily on the omentum ascertained to be of gestational origin by DNA polymorphism analysis. PMID- 15790468 TI - Palliative segmental duodenectomy for bleeding, erosive endometrial cancer. AB - BACKGROUND: Endometrial cancer is the fourth most common malignancy among women. Metastases typically involve the lungs, peritoneal surfaces of the abdominal cavity, and lymph nodes outside the pelvis. CASE: We report an unusually difficult complication of metastatic endometrial cancer occurring in a 57 year old woman with paraaortic nodal recurrence: refractory occult gastrointestinal bleeding. Repeated upper endoscopic evaluation revealed an extrinsic mass eroding into the third portion of the duodenum. Segmental duodenal resection was performed, and the GI tract was reconstructed with a side-to-side duodenojejunostomy. The postoperative course was uneventful, and the patient experienced durable relief from continued bleeding and impending obstruction. CONCLUSION: We review the role of palliative surgery for bleeding complications of advanced cancer and the alternatives for treating invasive extrinsic lesions of the duodenum. PMID- 15790469 TI - Primary ovarian large B-cell lymphoma in patient with juvenile rheumatoid arthritis treated with low dose Methotrexate. AB - BACKGROUND: Primary ovarian lymphoma is an extremely rare disease and limited count reports about it have been reported in the literature. Traditionally, patients with rheumatoid arthritis (RA) have increased risk of nodal and extranodal lymphoid malignancies such as non-Hodgkin's lymphoma (NHL). Recently, some studies have also reported association between patients with juvenile rheumatoid arthritis (JRA) treated with Methotrexate (MTX) and malignant lymphoma developing. CASE: We report a 17-year old JRA patient with primary ovarian diffuse large B-cell non-Hodgkin's lymphoma (NHL). The patient had seronegative (rheumatoid factor negative) poliarticular form of JRA and was receiving low dose weekly Methotrexate (MTX) during the past 2 years. Initial presentation was adnexial mass and chronic pelvic pain. The patient was treated with surgery and combined cytotoxic chemotherapy. CONCLUSION: In conclusion, because of increased lymphoid malignancy risk, ovarian masses in JRA patients should be carefully evaluated. PMID- 15790470 TI - A case of invasive squamous cell carcinoma on the surface of pedunculated cervical leiomyoma presenting an exophytic cervical cancer. AB - BACKGROUND: Cervical cancer on the mucosal surface of a pedunculated cervical leiomyoma has not been reported. CASE: A 54-year-old woman who had a bulky protruding cervical tumor with a history of a vaginal bleeding was diagnosed as having squamous cell cancer of the cervix. She received a total of 45 Gy (1.8 Gy daily using 15MV photons) external pelvic irradiation. However, on the follow-up MRI, the mass showed no response. The pedunculated exophytic mass was excised and the pathologic examination showed a leiomyoma covered by cervical stroma and squamous cell carcinoma. She then received additional intracavitary brachytherapy. CONCLUSION: This is the first case reported in the literature of squamous cell carcinoma on the surface of peduculated cervical leiomyoma that presented as an exophytic cervical tumor. PMID- 15790471 TI - Endometroid adenocarcinoma of the uterus with cardiac metastasis. A case report and six-year follow-up. AB - BACKGROUND: There are few reported cases of cardiac metastasis associated with endometrial cancer (EC) and no reports of long-term survival. We report a case of EC presenting with metastasis to the right ventricle resulting in right heart failure. CASE: A 61-year-old woman presented with a 10-day history of increasing pedal edema. Two-dimensional echocardiography (2DE) revealed a large echogenic mass within the right ventricle. She underwent exploration of the heart with debulking which revealed a metastatic poorly differentiated epithelial tumor. Further work-up with a dilation and curettage revealed a poorly differentiated EC. A total abdominal hysterectomy with pelvic and para-aortic lymhadenectomy was performed with disease confined to the uterus/cervix. She then received cardiac radiation with concurrent cisplatin, followed by pegylated liposomal doxorubicin (PLD) for 1 year and remained disease free through 6.5 years of follow-up. At 6.8 years, she died of a constrictive pericarditis with no evidence of disease. CONCLUSION: While experience with cardiac metastasis for gynecologic malignancy is limited, it appears that multimodality therapy affected a durable complete response however late complications of cardiac radiation ultimately lead to death. PMID- 15790472 TI - Steroid cell tumors, not otherwise specified (NOS), in an accessory ovary: a case report and literature review. AB - BACKGROUND: Accessory ovaries are uncommon congenital abnormalities of the female reproductive tract. Steroid cell tumors at accessory ovaries are proportionately even rare. CASE: This 30-year-old woman had an accessory ovarian tumor attached to the infundibulum of the right fallopian tube which was solid and gross pathologically and microscopically showed the appearances of steroid cell tumor, NOS, a benign entity. CONCLUSION: We report the first case of an accessory ovarian steroid cell tumor, NOS, which attached to the infundibulum of fallopian tube. PMID- 15790473 TI - Borderline endometrioid tumor arising in a paratubal cyst: a case report. AB - BACKGROUND: Paratubal cysts, generally known as hydatid cysts of Morgagni, are small round cysts attached by a pedicle to the fimbriated end of the tube. The following represents the first reported case of an endometrioid tumor of low malignant potential arising in a paratubal cyst. CASE REPORT: A 45-year-old nulliparous female was referred with a complex right adnexal mass on pelvic sonogram. She underwent laparoscopic bilateral ovarian cystectomy and partial right salpingectomy (for a 3-cm torsed paratubal cyst). All frozen-section diagnoses were benign; however, final pathology revealed a borderline tumor of low malignant potential of endometrioid type in the right paratubal cyst. The patient underwent extensive counseling regarding management options and decided to have a laparoscopic right salpingo-oophorectomy. Final pathologic analysis revealed no evidence of persistent borderline tumor. CONCLUSION: Paratubal cysts are very difficult to diagnose with sonography; therefore, their management should be approached as any other adnexal mass. Laparoscopic surgery is an option in the management of adnexal masses; however, rupture or puncture of masses should be avoided when possible to prevent potential tumor dissemination in the event of a malignancy. PMID- 15790474 TI - Optimal cytoreduction after combined resection and radiofrequency ablation of hepatic metastases from recurrent malignant ovarian tumors. AB - BACKGROUND: The role of radiofrequency ablation (RFA) in the treatment of hepatic metastases from recurrent ovarian tumors is undefined. CASE: Three patients with hepatic lesions from recurrent ovarian cancers underwent a combined partial hepatectomy with radiofrequency ablation (RFA) to achieve optimal tumor cytoreduction. Follow-up radiological studies as well as serial tumor markers are consistent with disease-free survival after 39, 13, and 9 months. CONCLUSION: These results demonstrate the feasibility and safety of RFA for metastatic ovarian lesions to the liver in patients previously deemed as poor or non surgical candidates, and suggest the potential for improvement in survival over unresected patients or in patients resected with residual disease. PMID- 15790475 TI - Post-transplant lymphoproliferative disorder of the cervix. AB - BACKGROUND: Post-transplant lymphoproliferative disorder involving the female genital tract is a rare event. CASE: A 67-year-old female status post orthotopic liver transplant 5 years previously for hepatitis B and sarcoidosis presented with vaginal bleeding. Endometrial biopsy revealed a high-grade malignant tumor with immunologic marker studies consistent with lymphoma. The patient underwent an exploratory laparotomy, modified radical hysterectomy, bilateral salpingo oophorectomy, bilateral selective pelvic and para-aortic lymphadenectomy, and omentectomy. Final pathology confirmed monomorphic B-cell post-transplantation lymphoproliferative disorder consistent with non-Hodgkin's B cell lymphoma confined to the endocervix and lower uterine segment. She remains recurrence free after 4 years with no adjuvant therapy. CONCLUSION: Post-transplant complications can present in the female reproductive organs. Gynecologic oncologists need to be aware of this disease process when treating patients for gynecologic symptoms after transplant surgery. PMID- 15790476 TI - Single vaginal ectopic ureter of fetal metanephric duct origin, ipsilateral kidney agenesis, and ipsilateral rudimentary uterine horn of the bicornuate uterus. AB - BACKGROUND: Congenital anomalies arising from the urogenital tracts are common. We experienced two cases of specific urogenital anomalies. CASE: Two Japanese women had a single ectopic ureter opening in the central wall of the vagina, ipsilateral kidney agenesis, and ipsilateral rudimentary uterine horn of the bicornuate uterus. One had squamous cell carcinoma of the uterine cervix, and the other had adenocarcinoma of the vagina arising from the ectopic ureter meatus, which was histologically confirmed as fetal metanephric duct remnant. One had vesico-vaginal fistula postoperatively, resulting from separating the ureter-like duct from the bladder. CONCLUSION: Concomitant occurrence of anomalies suggests the common genetic backgrounds, which are not well known. Clinically, the ectopic vaginal ureter is important as a possible background of vaginal adenocarcinoma, and information on the ureter-like duct course is required during surgery. PMID- 15790477 TI - Metachronous colorectal malignancies: "don't forget the neo vagina". A case report. AB - BACKGROUND: Carcinoma arising in a neovagina is rare. CASE: A patient with an adenocarcinoma arising from a neovagina constructed with use of a sigmoid segment. CONCLUSION: Transplanted intestinal tissue should not be overlooked, especially in patients with an increased risk for developing metachronous malignancies. PMID- 15790478 TI - Pseudo-Meigs syndrome and elevated CA125 associated with struma ovarii. AB - BACKGROUND: Struma ovarii is a rare form of ovarian neoplasm composed entirely and predominantly of thyroid tissue. The association of pseudo-Meigs syndrome, elevation of CA125 and hyperthyroidism to struma ovarii is a rare condition. CASE: We report an unusual presentation of a postmenopausal woman with benign struma ovarii associated with pseudo-Meigs syndrome, hypertiroidism, and elevated CA125 serum level, and a large complex right pelvic mass thereby mimicking an ovarian cancer. CONCLUSIONS: Struma ovarii is a rare cause of ascites, hydrothorax, elevated CA125, and hyperthyroidism. This rare condition should be considered in the differential diagnosis in patients with ascites and pleural effusions but with negative cytology. PMID- 15790479 TI - Neoadjuvant combined chemotherapy followed by external whole pelvic irradiation in two cases of primary extranodal non-Hodgkin's lymphoma of the uterine cervix. AB - BACKGROUND: Primary extranodal lymphomas of the genital tract are rare. CASES: As there is no current consensus in its management, we present two further cases and their treatment with neoadjuvant chemotherapy, followed by radiation therapy. A radical hysterectomy with bilateral pelvic lymphadenectomy was performed after primary treatment in one case. Clinical response was complete in both cases and pathological response was documented in one. CONCLUSIONS: Complete response of these lymphoid neoplasms can be achieved by neoadjuvant chemotherapy followed by external irradiation. PMID- 15790480 TI - Ascites, pleural effusion, and CA 125 elevation in an SLE patient, either a Tjalma syndrome or, due to the migrated Filshie clips, a pseudo-Meigs syndrome. AB - BACKGROUND: The combination ascites, pleural effusion, and elevated CA 125 are usually associated with a malignancy. CASE: A 38-year-old SLE patient consulted her physician for shortness of breath. On clinical examination, she had a tender abdomen and reduced breathing sounds. X-ray and computed tomography of the chest showed pleural effusion. An adjustment of her SLE maintenance therapy was performed. Vaginal ultrasound and computed tomography of the abdomen revealed massive ascites and an intracavitair myoma of 2 cm, but no obvious mass in the pelvis. CA 125 was 887 U/ml. A laparoscopy was performed showing ascites and 2 Filshie clips embedded in the peritoneum of the vesicouterine pouch, but no sign of malignancy. Both clips were removed. The cytology of the aspirated ascites showed sings of acute inflammation. Within 10 weeks, the pleural effusion was resolved and the CA 125 normalized. CONCLUSION: The combination of ascites, pleural effusion, CA 125 elevation, and no tumor in an SLE patient is either a Tjalma syndrome or due to the migrated Filshie clips a pseudo-Meigs syndrome. PMID- 15790481 TI - Umbilical metastasis after laparoscopic retroperitoneal paraaortic lymphadenectomy for cervical cancer: a true port-site metastasis? AB - BACKGROUND: We present a case of umbilical metastasis after laparoscopic retroperitoneal paraaortic lymphadenectomy for cervical cancer. CASE: A 59-year old woman with stage IIIB cervical adenocarcinoma underwent laparoscopic paraaortic lymphadenectomy as well as a conventional laparoscopy to assess the presence of peritoneal carcinomatosis. The pathologic examination revealed metastasis in one paraaortic node. Peritoneal cytology proved negative for malignant cells. Seven months after completion of chemoradiotherapy, the patient presented a 2.5-cm umbilical tumor involving the trocar tract together with recurrence of the cervical mass. Histological examination of the excised umbilical mass showed recurrence of the cervical adenocarcinoma, with strong peritumoral CD31 immunocytochemical expression. CONCLUSION: The peritumoral increase in microvessel density and strong CD31 positivity suggests angiogenesis as a potential factor lead to seeding of tumor cells at the umbilical port. PMID- 15790482 TI - Pediatric radical abdominal trachelectomy for cervical clear cell carcinoma: a novel surgical approach. AB - INTRODUCTION: Clear cell carcinoma of the cervix and vagina is rare in the pediatric population. Many of these tumors are associated with prior intrauterine diethylstilbestrol (DES) exposure. All DES-associated tumors are believed to contain microsatellite instability (MI). Historically, the recommended treatment is radical hysterectomy and pelvic lymphadenectomy, which result in infertility in all cases. Radical abdominal or vaginal trachelectomy and pelvic lymph node dissection is a new technique utilized in adult women with early cervical cancer who wish to retain fertility. This novel approach is also pertinent to the pediatric patient and is described in this report. A molecular analysis is also performed to determine if these are DES-associated tumors. METHODS: Due to the narrow vaginal anatomy in pediatric patients, a vaginal approach is not possible, and an abdominal approach is performed. The resection includes the cervix, upper vagina, parametrium, and paracolpos. Pelvic lymphadenectomy is performed in a similar manner to the adult patient. Matched pairs of normal and tumor DNA from both cases were examined for evidence of MI using a consensus panel of microsatellite markers. RESULTS: Two girls aged 6 and 8 years and without history of DES exposure presented with vaginal bleeding. Vaginoscopy revealed cervical polyp in both cases. Biopsies demonstrated clear cell cancer stage IB1 in both patients. They underwent radical abdominal trachelectomy and bilateral pelvic lymph node dissection along with anastomosis of uterine isthmus to upper vagina. Intraoperative frozen-section analysis confirmed negative uterine and vaginal margins. No adjuvant treatment was given and both girls remain disease free. Neither tumor showed evidence for MI, confirming that these are not DES associated tumors. CONCLUSION: To our knowledge, this is the first report of radical abdominal trachelectomy in the pediatric age group and it is likely to include the youngest patient with clear cell carcinoma of the uterine cervix not associated with DES exposure. This novel approach is feasible and appears safe in the pediatric age group. PMID- 15790483 TI - 16,090: the 2004 estimated U.S. mortality from ovarian cancer. PMID- 15790484 TI - Re: "Cultures of ovarian surface epithelium from women with and without a hereditary predisposition to develop female adnexal carcinoma". PMID- 15790486 TI - Detection of an activating c-kit mutation by real-time PCR in patients with anaphylaxis. AB - A well-characterised gain-of-function point mutation within exon 17 of the c-kit proto-oncogene known as Asp816Val is present in patients with mastocytosis. Activation of mast cells through this receptor primes them for IgE-dependent activation, and patients with mastocytosis are at increased risk of anaphylaxis. We hypothesised that the Asp816Val mutation is associated with a history of anaphylaxis in the general population. A mismatch amplification real-time PCR assay was developed and validated to test for the Asp816Val mutation. Subjects were recruited to four subject groups: normal non-atopics, atopics without anaphylaxis, food-induced anaphylactics and non-food anaphylactics. Blood samples collected from forty subjects were tested for the presence of Asp816Val. Thirteen subjects were found to carry the mutation; normals (2/9), atopics (2/10), food anaphylactics (5/11) and non-food anaphylactics (4/10). Statistical analysis of the data determined that there was no significant difference between the numbers of subjects found to carry the Asp816Val mutation in each of the groups although a trend towards an increased occurrence in anaphylactics was observed. In summary, the hypothesis that the presence of the Asp816Val mutation is linked to the occurrence of anaphylaxis was not supported, but interestingly, we have shown for the first time Asp816Val may occur more frequently than previously reported within the general population. PMID- 15790487 TI - Age-dependent sensitivity of Big Blue transgenic mice to the mutagenicity of N ethyl-N-nitrosourea (ENU) in liver. AB - The incidence of childhood cancer is increasing and recent evidence suggests an association between childhood cancer and environmental exposure to genotoxins. In the present study, the Big Blue transgenic mouse model was used to determine whether specific periods in early life represent windows of vulnerability to mutation induction by genotoxins in mouse liver. Groups of mice were treated with single doses of 120 mg N-ethyl-N-nitrosourea (ENU)/kg body weight or the vehicle either transplacentally to the 18-day-old fetus or at postnatal days (PNDs) 1, 8, 15, 42 or 126; the animals were sacrificed 6 weeks after their treatment. The cII mutation assay was performed to determine the mutant frequencies (MFs) in the livers of the mice. Liver cII MFs for both sexes were dependent on the age at which the animals were treated. Perinatal treatment with ENU (either transplacental treatment to the 18-day-old fetus or i.p. injection at PND 1) induced relatively high MFs. However, ENU treatment at PNDs 8 and 15 resulted in the highest mutation induction. The lowest mutation induction occurred in those animals treated as adults (PND 126). For instance, the cII MF for the PND 8 female group was 646 x 10(-6) while the MF for female adults was only 145 x 10( 6), a more than 4-fold difference. Molecular analysis of the mutants found that A:T-->T:A transversions and A:T-->G:C transitions characterized the pattern of mutations induced by ENU in both the neonate and adult mice, while the predominate type of mutation in the controls was G:C-->A:T. The results indicate that mouse liver is most sensitive to ENU-induced mutation during infancy. This period correlates well with the age-dependent sensitivity to carcinogenicity in mouse liver, suggesting that mutation is an important rate-limiting factor for age-related carcinogenesis. PMID- 15790488 TI - The cell cycle phases of DNA damage and repair initiated by topoisomerase II targeting chemotherapeutic drugs. AB - Although cytostasis and cytotoxicity induced by cancer chemotherapy drugs targeting topoisomerase II (topoII) arise in specific cell cycle phases, it is unknown whether the drug-initiated DNA damage triggering these responses, or the repair (reversal) of this damage, differs between cell cycle phases or between drug classes. Accordingly, we used a flow cytometric alkaline unwinding assay to measure DNA damage (strand breakage (SB)) and SB repair in each cell cycle compartment of human cancer cell lines treated with clinically relevant concentrations of doxorubicin, daunomycin, etoposide, and mitoxantrone. We found that treated HeLa and A549 cells exhibited the greatest SB in G2/M phase, the least in G1 phase, and generally an intermediate amount in S phase. The cell cycle phase specificity of the DNA damage appeared to be predictive of the cell cycle phase of growth arrest. Furthermore, it appeared to be dependent on topoIIalpha expression as the extent of SB did not differ between cell cycle compartments in topoIIalpha-diminished A549(VP)28 cells. HeLa cells were apparently unable to repair doxorubicin-initiated SB. The rate of repair of etoposide-initiated SB in HeLa cells and of mitoxantrone-initiated SB in HeLa and A549 cells was similar in each cell cycle compartment. In A549 cells, the rate of repair of doxorubicin and etoposide-initiated SB differed between cell cycle phases. Overall, these results indicate that the cell cycle phase specificity of cytostasis and cytotoxicity induced in tumor cells by topoII-targeting drugs may be directly related to the cell cycle phase specificity of the drug-initiated DNA damage. Analysis by cell cycle compartment appears to clarify some of the intercellular heterogeneity in the extent of drug-initiated DNA damage and cytotoxicity previously observed in cancer cells analyzed as a single population; this approach might be useful in resolving inconsistent results reported in investigations of tumor cell topoII content versus response to topoII-targeting drugs. PMID- 15790489 TI - Application of a two-stage Syrian hamster embryo cell transformation assay to cigarette smoke particulate matter. AB - The induction of transformation in Syrian hamster embryo (SHE) cells is a multifactorial process, in comparison to endpoints induced in in vitro genotoxicity assays such as Ames, mouse lymphoma and cytogenetics [Y. Berwald, L. Sachs, In vitro cell transformation with chemical carcinogens, Nature (London) 200 (1963) 1182-1184]. Furthermore, a number of non-genotoxic carcinogens and promoters such as clofibrate and diethylhexylphthalate, have been positively identified in this assay, while giving false negative results in traditional genotoxicity assays [H. Yamasaki, J. Ashby, M. Bignami, W. Jongen, K. Linnainmaa, R.F. Newbold, G. Nguyen-Ba, S. Parodi, E. Rivedal, D. Schiffmann, J.W.I.M. Simons, P. Vasseur, Nongenotoxic carcinogens: development of detection methods based on mechanisms: a European project, Mutat. Res. 353 (1996) 47-63]. A high concordance between results obtained in this assay when compared with rodent carcinogenesis bioassays has also been noted [R.J. Isfort, G.A. Kerckaert, R.A. LeBoeuf, Comparison of the standard and reduced pH Syrian hamster embryo (SHE) in vitro cell transformation assays to predict the carcinogenic potential of chemicals, Mutat. Res. 356 (1996) 11-63]. Carcinogenesis is known to be a multistage process, with agents potentially acting at each stage. Specifically, mouse skin painting experiments established that tumour induction could be mechanistically divided into two distinct phases, termed initiation and promotion. Initiation, is defined as the stage at which a normal cell is converted to a latent tumour cell, followed by promotion where the latent tumour cell progresses to a tumour [W.F. Friedwald, P. Rous, The initiating and promoting elements in tumour production: analysis of the effects of tar, benzpyrene and methylcholanthrene on rabbit skin, J. Exp. Med. 80 (1944) 101 125]. A protocol for the pH 6.7 SHE transformation assay has been developed which allows separation of cell transformation process into two phases, potentially analogous to initiation and promotion in vivo. This allows chemicals found to be positive in the traditional SHE cell transformation assay to be further classified as initiators or promoters. Following validation with known initiators, benzo(a)pyrene and N-methyl-N'-nitro-N-nitrosoguanidine and promoters, 12-O-tetradecanoyl-phorbol-13-acetate and phenobarbitone, the two stage model was applied to cigarette smoke particulates which was found to act both at the initiation and promotion stage of cell transformation. PMID- 15790490 TI - Inhibition of urethane-induced genotoxicity and cell proliferation in CYP2E1-null mice. AB - Urethane is a multi-site animal carcinogen and was classified as "reasonably anticipated to be a human carcinogen." Urethane is a fermentation by-product and found at appreciable levels in alcoholic beverages and foods such as bread and cheese. Recent work in this laboratory demonstrated for the first time that CYP2E1 is the principal enzyme responsible for urethane metabolism. The current studies were undertaken to assess the relationships between CYP2E1-mediated metabolism and urethane-induced genotoxicity and cell proliferation as determined by induction of micronucleated erythrocytes (MN) and expression of Ki-67, respectively, using CYP2E1-null and wild-type mice. Urethane was administered at 0 (vehicle), 1, 10, or 100mg/kg/day (p.o.), 5 days/week for 6 weeks. A significant dose-dependent increase in MN was observed in wild-type mice; however, a slight increase was measured in the MN-polychromatic erythrocytes in CYP2E1-null mice treated with 100mg/kg. A significant increase in the expression of Ki-67 was detected in the livers and the lungs (terminal bronchioles, alveoli, and bronchi) of wild-type mice administered 100mg urethane/kg in comparison to controls. In contrast, CYP2E1-null mice administered this dose exhibited negligible alterations in Ki-67 expression in the livers and lungs compared to controls. Interestingly, while Ki-67 expression in the forestomach decreased in wild-type mice, it increased in CYP2E1-null mice. Subsequent comparative metabolism studies demonstrated that total urethane-derived radioactivity in the plasma, liver, and lung was significantly higher in CYP2E1-null versus wild-type mice and un-metabolized urethane constituted greater than 83% of the radioactivity in CYP2E1-null mice. Un-metabolized urethane was not detectable in the plasma, liver, and lung of wild-type mice. In conclusion, these data demonstrated that CYP2E1-mediated metabolism of urethane, presumably via epoxide formation, is necessary for the induction of genotoxicity, and cell proliferation in the liver and lung of wild-type mice. PMID- 15790491 TI - Homologous recombination in plants is temperature and day-length dependent. AB - Homologous recombination (HR) as a strand break repair mechanism was shown to be influenced by various factors. The balance of different vitamins, macro- and microelements, light spectrum, sodium chloride concentration as well as various environmental mutagens were shown to influence the frequency of HR. In this paper we analysed the influence of temperature (4, 22, and 32 degrees C) and day/night duration on the genome stability of plants. We analyzed the HR frequency in transgenic Arabidopsis thaliana plants carrying beta-glucuronidase based homologous recombination substrate. To find the recombination rate (RR), we related the HR frequency to the number of genomes present in plants grown under different conditions. The RR was also standardized to the transcription activity of the transgene. We found RR to be higher in plants grown at suboptimal temperatures (either 4 or 32 degrees C) as compared to plants grown at 22 degrees C. This negatively correlated with the plant metabolic rate and positively correlated with concentration of peroxide produced by plant. In contrast, the RR in plants grown at different day length (8-24 h) was the lowest in plants grown at the longest day (24 h) and highest in the plants grown at the shortest day (8 h). Over 15-fold difference in the RR between plants grown at the shortest and the longest day was observed. Such a difference in recombination rate was primarily due to the higher transgene activity and higher endoreduplication levels in plants grown at longer days. Our data suggests that even "moderate" changes of environmental conditions may have a significant effect on plant genome stability. PMID- 15790492 TI - Induction of petite mutants in yeast Saccharomyces cerevisiae by the anticancer drug dequalinium. AB - Dequalinium (DEQ), a drug with both antimicrobial and anticancer activity, induced the formation of petite (respiration-deficient) mutants in the yeast Saccharomyces cerevisiae. DEQ was found to be approximately 50-fold more potent than ethidium bromide (EB) at inducing petites. Analysis of the DEQ-induced petite mutants indicated a complete loss of mitochondrial DNA (<1 copy/cell). Prior to the loss of mtDNA, DEQ caused cleavage of the mtDNA into a population of fragments 30-40kbp in size suggesting that this drug causes petites by inducing a breakdown of mtDNA. The selective effect of DEQ on yeast mtDNA may underlie the antifungal activity of this chemotherapeutic agent. PMID- 15790493 TI - Genotoxicity and metabolism of the source-water contaminant 1,1-dichloropropene: activation by GSTT1-1 and structure-activity considerations. AB - 1,1-Dichloropropene (1,1-DCPe) is a contaminant of some source waters used to make drinking water. Because of this and the fact that no toxicological data were available for this compound, which is structurally similar to the rodent carcinogen 1,3-dichloropropene (1,3-DCPe), 1,1-DCPe was placed on the Contaminant Candidate List of the US Environmental Protection Agency. Consequently, we have performed a hazard characterization of 1,1-DCPe by evaluating its mutagenicity in the Salmonella assay and its DNA damaging (comet assay) and apoptotic (caspase assay) activities in human lymphoblastoid cells. In Salmonella, 1,1-DCPe was not mutagenic in strains TA98, TA100, TA1535, or TA104 +/-S9 mix. However, it was clearly mutagenic in strain RSJ100, which expresses the rat GSTT1-1 gene. 1,1 DCPe did not induce DNA damage in GSTT1-1-deficient human lymphoblastoid cells, and it induced apoptosis in these cells only at 5 mM. Consistent with its mutagenesis in RSJ100, 1,1-DCPe reacted with glutathione (GSH) in vitro, suggesting an addition-elimination mechanism to account for the detected GSH conjugate. 1,1-DCPe was approximately 5000 times more mutagenic than its ethene congener 1,1-dichloroethylene (1,1-DCE or vinylidene chloride). Neither 1,1-DCE nor 1,3-DCPe showed enhanced mutagenicity in strain RSJ100, indicating a lack of activation of these congeners by GSTT1-1. Thus, 1,1-DCPe is a base-substitution mutagen requiring activation by GSTT1-1, possibly involving the production of a reactive episulfonium ion. This bioactivation mechanism of 1,1-DCPe is different from that of its congeners 1,1-DCE and 1,3-DCPe. The presence of 1,1-DCPe in source waters could pose an ecological or human health risk. Occurrence data for 1,1-DCPe in finished drinking water are needed to estimate human exposure to, and possible health risks from, this mutagenic compound. PMID- 15790494 TI - Mutator activity and specificity of Escherichia coli dnaQ49 allele--effect of umuDC products. AB - The high fidelity of DNA replication in Escherichia coli is ensured by the alpha (DnaE) and epsilon (DnaQ) subunits of DNA polymerase providing insertion fidelity, 3'-->5' exonuclease proofreading activity, and by the dam-directed mismatch repair system. dnaQ49 is a recessive allele that confers a temperature sensitive proofreading phenotype resulting in a high rate of spontaneous mutations and chronic induction of the SOS response. The aim of this study was to analyse the mutational specificity of dnaQ49 in umuDC and DeltaumuDC backgrounds at 28 and 37 degrees C in a system developed by J.H. Miller. We confirmed that the mutator activity of dnaQ49 was negligible at 28 degrees C and fully expressed at 37 degrees C. Of the six possible base pair substitutions, only GC-->AT transitions and GC-->TA and AT-->TA transversions were appreciably increased. However, the most numerous mutations were frameshifts, -1G deletions and +1A insertions. All mutations which increased in response to dnaQ49 damage were to a various extent umuDC-dependent, especially -1G deletions. This type of mutations decreased in CC108dnaQ49DeltaumuDC to 10% of the value found in CC108dnaQ49umuDC+ and increased in the presence of plasmids producing UmuD'C or UmuDC proteins. In the recovery of dnaQ49 mutator activity the plasmid harbouring umuD'C genes was more effective than the one harbouring umuDC. Analysis of mutational specificity of pol III with defective epsilon subunit indicates that continuation of DNA replication is allowed past G:T, C:T, T:T (or C:A, G:A, A:A) mismatches but does not allow for acceptance of T:C, C:C, A:C (or A:G, G:G, T:G) (the underlined base is in the template strand). PMID- 15790495 TI - A novel selectable system for detecting expansion of CAG.CTG repeats in mammalian cells. AB - CAG.CTG repeat expansions cause more than a dozen neurodegenerative diseases in humans. To define the mechanism of repeat instability in mammalian cells we developed a selectable assay to detect expansions of CAG.CTG triplet repeats in Chinese hamster ovary (CHO) cells. We showed previously that long tracts of CAG.CTG repeats, embedded in an intron of the APRT gene, kill expression of the gene, rendering the cells APRT-. By contrast, tracts with fewer than 34 repeats allow sufficient expression to give APRT+ cells. Although it should be possible to use APRT+ cells with short repeats to assay for expansion events by selecting for APRT- cells, we find that APRT+ cells with 31 repeats are not killed by the standard APRT- selection protocol, most likely because they produce too little Aprt to incorporate sufficient 8-azaadenine into their adenine pool. To overcome this problem, we devised a new selection, which increases the proportion of the adenine pool contributed by the salvage pathway by partially inhibiting the de novo pathway. We show that APRT- CHO cells with 61 or 95 CAG.CTG repeats survive this selection, whereas cells with 31 repeats die. Using this selection system, we can select for expansion to as few as 39 repeats. Thus, this assay can monitor expansions across the critical boundary from the longest lengths of normal alleles to the shortest lengths of disease alleles. PMID- 15790496 TI - Frequent retention of heterozygosity for point mutations in p53 and Ikaros in N ethyl-N-nitrosourea-induced mouse thymic lymphomas. AB - In agreement with Knudson's two-hit theory, recent findings indicate that the inactivation of tumor suppressor genes is not only mediated by the loss of function but also by the dominant-negative or gain-of-function activity. The former generally accompanies loss of a wild-type allele whereas in the latter a wild-type allele is retained. N-Ethyl-N-nitrosourea (ENU), which efficiently induces point mutations, reportedly leads to the development of tumors by activating ras oncogenes. Little is known about how ENU affects tumor suppressor genes and, therefore, we examined ENU-induced mutations of p53 and Ikaros in thymic lymphomas and compared these with mutations of Kras. In addition, loss of heterozygosity was examined for chromosome 11 to which both p53 and Ikaros were mapped. The frequency of point mutations in p53 and Ikaros was 30% (8/27) and 19% (5/27), respectively, comparable to that observed in Kras (33%: 9/27). In total, 14 of the 27 thymic lymphomas examined (52%) harbored mutations in at least one of these genes. One Ikaros mutation was located at the splice donor site, generating a novel splice isoform lacking zinc finger 3, Ik (F3del). Interestingly, 90% (10/11) of the tumors with point mutations retained wild-type alleles of p53 and Ikaros. Sequence analysis revealed that the most common nucleic acid substitutions were T>A (4/8) in p53, T>C (4/5) in Ikaros and G>A/T (8/9) in Kras, suggesting that the spectrum of mutations was gene dependent. These results suggest that point mutations in tumor suppressor genes without loss of the wild-type allele play an important role in ENU-induced lymphomagenesis. PMID- 15790497 TI - Visible light (>395 nm) causes micronuclei formation in mammalian cells without generation of cyclobutane pyrimidine dimers. AB - Solar radiation gives rise to DNA damage in mammalian cells not only directly by excitation of DNA, which generates predominantly pyrimidine dimers, but also indirectly by the excitation of endogenous photosensitizers, which causes oxidative DNA modifications. The latter mechanism has a low quantum yield, but it is the only one proceeding in the visible range of the spectrum. To investigate its relevance for the genotoxicity of sunlight, we have analysed the generation of micronuclei associated with the induction of oxidative DNA damage by visible light in melanoma cells and primary human skin fibroblasts. Similar yields of light-induced oxidative DNA base modifications sensitive to the repair glycosylase Fpg (7,8-dihydro-8-oxoguanine and other oxidative purine modifications) were observed in the normal fibroblasts and the malignant melanoma cells of the same donor. When irradiations were carried out at intervals to compensate for a photodecomposition of the endogenous chromophore, a significant generation of micronuclei was observed in both cell types. Cyclobutane pyrimidine dimers could be excluded to be responsible for the micronuclei induction at wavelengths >395 nm. Experiments with a cut-off filter indicate that the ratio of pyrimidine dimers and Fpg-sensitive oxidative modifications in irradiated cells not only reflects the relative contributions of direct and indirect mechanisms, but is also similar to the ratio by which the two mechanisms contribute to the generation of the micronuclei. The results suggest that indirectly generated oxidative DNA modifications can contribute significantly to the adverse effects of sunlight. PMID- 15790498 TI - Involvement of nucleotide excision repair (NER) system in repair of mono ADP ribosylated dG adducts produced by pierisin-1, a cytotoxic protein from cabbage butterfly. AB - Pierisin-1, a cytotoxic protein from the cabbage butterfly (Pieris rapae), induces apoptosis in mammalian cell lines. Binding of its C-terminal region to glycosphingolipid Gb3 and Gb4 receptors on cell membrane is necessary for incorporation into cells, while the N-terminal polypeptide catalyzes transfer of the ADP-ribose moiety of NAD at N2 of dG in DNA. Resulting DNA adducts cause mutation if they are present at low levels. If the DNA damage is more severe, the cells undergo apoptosis. In the present study, we examined the repair system for ADP-ribosylated dG adducts using nucleotide excision repair (NER) mutants of Chinese hamster ovary (CHO) cell lines. Pierisin-1 showed cytotoxic effects in all cases: IC50 values of them were; 650 ng/ml for AA8 (wild), 230 ng/ml for UV5, 190 ng/ml for UV20, 260 ng/ml for UV41, and 240 ng/ml for UV135. Thus, wild-type AA8 proved most resistant to pierisin-1-induced cytotoxicity. When these CHO cell lines were treated with pierisin-1, the adduct levels of ADP-ribosylated dG increased to 2.5-4.8/10(5) nucleotides time-dependently in all cell lines at 12 h. After removal of pierisin-1, the adduct levels remained constant or increased to 4-14/10(5) nucleotides in all NER mutant cells (UV5, UV20, UV41, UV135), while those rapidly decreased to 0.27/10(5) nucleotides in the repair proficient AA8 cells for 24 h. From these results, it is suggested that the NER system is involved in the repair of ADP-ribosylated dG adducts in DNA. PMID- 15790499 TI - HPRT gene alterations in umbilical cord blood T-lymphocytes in newborns of mothers exposed to tobacco smoke during pregnancy. AB - Prenatal exposure to tobacco smoke has been associated with an increased risk of pediatric malignancies, yet the transplacental induction of genetic alterations by tobacco smoke carcinogens and their implication to childhood diseases remain poorly understood. We characterized mutations in the HPRT gene in umbilical cord blood T-lymphocytes of self-reported 103 never-smoking mothers and 104 smoking mothers (54 mothers smoked throughout and 50 mothers quit smoking during pregnancy). The results showed the illegitimate V(D)J recombinase-mediated deletion of HPRT exons 2-3 was the most prominent alteration occurring in 48.2% (26/54) of mutants from neonates of the smoking mothers who smoked during pregnancy, compared with 28.0% (14/50) from those of smoking mothers who quit smoking during pregnancy (p=0.035, Fisher's exact test), 34.9% (36/103) from never-smoking mothers (p=0.08), or 32.7% (50/153) of those of neonates born from the latter two groups of mothers combined (p=0.043). There was no significant difference in the frequency of this deletion between neonates of the never smoking mothers and the smoking mothers who quit smoking during pregnancy (34.9% versus 28.0%, respectively, p=0.39). The results show an increase in illegitimate V(D)J recombinase-mediated deletion of HPRT exons 2-3 in cord blood T-lymphocytes of newborns of mothers who smoked during pregnancy, compared with the group of mothers who did not smoke during pregnancy, implying an increase in illegitimate V(D)J recombinase-mediated alteration, a genetic recombination event associated with childhood malignancies, may be induced in utero during pregnancy by maternal exposure to tobacco smoke-derived genotoxicants. PMID- 15790500 TI - Interaction of hyperbaric oxygen, nitric oxide, and heme oxygenase on DNA strand breaks in vivo. AB - Hyperbaric oxygen (HBO), e.g. pure oxygen breathing at supra-atmospheric pressures, represents a well-suited model for investigating oxidative stress induced DNA damage as well as protective mechanisms. While the induction of heme oxygenase-1 (HO-1) seems to be crucial for this protection against this DNA damage, the role of nitric oxide (NO) remains unclear. HO-1 expression is a major regulator of the inducible NO synthase (iNOS), and therefore we investigated the effect of the interaction between HBO, NO, and HO-1 on DNA damage. Prior to exposure to HBO (3 h at 3 bar ambient pressure) rats randomly received vehicle (HBO alone, 1 mL 0.9% saline, n=8), the NO donor molsidomine (SIN-10, 40 mg/kg, n=8) or the HO-1 blocker tin-mesopophyrin (Sn-MP, 50 micromol/kg, n=8). Additional groups received SIN-10 without exposure to HBO, i.e. breathing air under normobaric conditions for 3h (SIN-10 alone, 40 mg/kg, n=6), vehicle without HBO (negative controls, n=6), and ethylmethanesulfonate without HBO (EMS, 200 mg/kg) (positive controls n=4). Immediately after the 3 h HBO or air breathing period blood was analysed for DNA strand breaks (tail moment in the alkaline comet assay) and nitrite+nitrate (chemoluminescence). Whereas the tail moment was ten-fold higher after EMS than in the negative controls, there was no effect of HBO nor SIN-10 alone. Together with HBO, pretreatment with SIN-10 doubled the tail moment, and Sn-MP increased it by 50%. In contrast to Sn-MP or HBO alone, SIN-10 resulted in a five-fold increase of nitrite+nitrate concentrations. We conclude that both HO-1 blockade and excess NO release promote DNA damage during HBO exposure in vivo. The effect of HO-1 inhibition is probably independent of the regulatory function of HO-1 for iNOS. PMID- 15790501 TI - Can one assess genotoxic and carcinogenic risk of asbestos without mentioning ingested fibres? PMID- 15790503 TI - Immune surveillance and anti-tumor immune responses: an anatomical perspective. AB - The development of adaptive immune responses against infectious agents relies on the initiation of antigen specific immune responses in secondary lymphoid organs and on the migration of effector cells at the site of infection. Similarly, the development of anti-tumor immunity depends on the recognition of tumor-derived antigens by specific lymphocytes in the context of the lymphoid tissues and on the re-localisation of the cells to the site of cell transformation. Here, we will review the preclinical studies, which have defined the spatial and temporal organisation of anti-tumor immunity, and discuss the implications of these findings in active immunotherapy. PMID- 15790504 TI - Immunophenotyping of acute leukaemias. AB - Progress in the management and understanding of acute leukaemia can only be obtained if these diseases are thoroughly investigated, both clinically and with a series of biological tools. This alone has made and still will make possible the identification of prognostic factors and of useful markers for the follow-up of patients in remission. Among the variety of approaches of acute leukaemia definition, immunophenotyping has taken over the past 25 years a predominant and now well-defined place, although room is left for further improvement. In this review, the current state-of-the-art of immunophenotyping of acute leukaemias will be replaced in the context of physiological leukocyte maturation. The recognized classifications and recommended immunophenotyping panels will then be discussed, and the clinical relevance of several key features will be presented. Finally, more recent openings for the use of immunophenotyping will be evoked. PMID- 15790505 TI - A causal link between lymphopenia and autoimmunity. AB - It is well recognized that the composition of the mature T cell population is subject to strict homeostatic control. The TCR repertoire and relative proportions of various T cell subsets are established in the thymus, and continue to be shaped and regulated in the periphery. As the thymic function declines, peripheral homeostatic mechanisms assume increasing importance. Indeed, loss of thymic function does not lead to progressive decline of T cell numbers because peripheral mechanisms ensure that the size of the T cell population is maintained due to proliferation of residual cells. However, our current understanding of the basic mechanisms of 'homeostatic' or lymphopenia-induced proliferation suggests that this drive to maintain population size may be accompanied by loss of TCR diversity and emergence of auto-reactive effector T cells. This prediction is supported by experimental and clinical evidence. This consideration is important because lymphopenia is seen commonly in clinical practice as a consequence of viral infections, or medical treatment of cancer, autoimmunity, and graft rejection. Lymphopenia may be a simple link between viral infections and autoimmunity, and may be one reason for common failure of very potent, but non specific, immunosuppressive drugs in current clinical use. PMID- 15790506 TI - Differential signalling during B-cell maturation. AB - The molecular mechanism by which the antigen receptors (BCR) on B cells can elicit differential maturation state-specific responses is one of the central problems in B-cell differentiation yet to be resolved. Indeed, many of the early signalling events detected following BCR ligation, such as activation of protein tyrosine kinases (PTK), phospholipase C (PLC), phosphoinositide-3-kinase (PI 3K), protein kinase C (PKC) and the RasMAPK (mitogen activating protein kinase) signalling cascades are observed throughout B-cell maturation. However, it is becoming clear that the differential functional responses of these BCR-coupled signals observed during B-cell maturation are dependent on a number of parameters including signal strength and duration, subcellular localisation of the signal, maturation-restricted expression of downstream signalling effector elements/isoforms and modulation of signal by co-receptors. Thus, the combined signature of BCR signalling is likely to dictate the functional response and act as a developmental checkpoint for B-cell maturation. PMID- 15790507 TI - TNF-alpha, TGF-beta and NO relationship in sera from tuberculosis (TB) patients of different severity. AB - Tuberculosis (TB) is the main cause of death by infection diseases worldwide. Considering that NO, TNF-alpha and TGF-beta participate a great deal in TB immunopathogenesis, we wished to analyse whether these mediators showed some relationship with the degree of pulmonary affectation. The sample comprised 29 TB (HIV-), inpatients with mild-moderate (n = 10) or advanced (n = 19) newly diagnosed disease, together with 12 healthy controls HCo. Serum nitrite was assessed by reducing nitrate to nitrite, and further measured by the Griess reaction. Levels of TNF-alpha and TGF-beta were determined by ELISA (R&D Systems). Serum levels of TNF-alpha were significantly higher in the advanced TB cases if compared with HCo, (p < 0.05 ) and from values of Mild-Moderate TB patients (p < 0.05). Serum levels of TGF-beta from advanced TB patients have increased values if compared with Hco (p < 0.005) and Mild-Moderate patients (p < 0.05). These values were also significantly different from Mild-Moderate cases + HCo (p = 0.01) Advanced TB patients had significantly reduced nitrite levels compared with those of Mild-Moderate patients and HCo (p < 0.002). Taken as a whole NO-derived metabolites in TB patients (M-M and Advanced cases) remained lower than values in HCo (p = 0.005) A negative correlation was found when comparing the two cytokines with nitrites(r = -0.44 ).TGF-beta and TNF-alpha were positively correlated (r = 0.44, p < 0.01), 0.44, p < 0.01. In synthesis, the inverse correlation found between both cytokines concentrations and NO levels in TB patients may be viewed as a consequence of a more predominant TGF-beta effect. PMID- 15790508 TI - Engineering of human complement component C3 for catalytic inhibition of complement. AB - As a novel therapeutic approach in complement-mediated pathologies, we recently developed a human C3 derivative capable of obliterating functional complement by a catalytic, non-inhibitory mechanism. In this derivative, the C-terminal region of hC3 was substituted by a 275 amino acid sequence derived from the corresponding sequence of cobra venom factor (CVF), a complement-activating C3b homologue from snake venom. In this study, we replaced shorter C-terminal sequences of hC3 by corresponding CVF sequences to further reduce potential immunogenicity and to identify domains essential for the formation of functionally stable C3 convertases. In one of these derivatives that is still capable of obliterating functional complement in vitro, the non-human portion could be reduced to a small domain located in the C-terminus of different complement proteins. This conserved NTR/C345C motif is known to be involved in assembly of different convertases of the complement system. These results suggest a major role of the C345C domain in the regulation of the half-life of the C3 convertase. Moreover, its overall identity of 96% to human C3 renders this derivative a promising candidate for therapeutic intervention in complement mediated pathologies. PMID- 15790509 TI - T-cell activation is accompanied by an ubiquitination process occurring at the immunological synapse. AB - The immunological synapse (IS) is a specialized signaling area formed at the contact site between T-cells and antigen-presenting cells (APC), where sustained engagement and signaling of TCR and accessory molecules occur. A key feature of T cell antigen recognition is that the process of TCR/peptide-MHC interaction is self-limited by the internalization and degradation of triggered TCR and recruited signaling components. The mechanism of signaling component degradation involves their ubiquitination and targeting for degradation. Yet, the relationship between the ubiquitination process and TCR signaling as well as the cellular localization of TCR-induced ubiquitination are still elusive. In the present work, we visualize for the first time ubiquitination at the TCR signaling area. We show an enrichment of ubiquitin staining in TCR/CD3 caps in T lymphocytes stimulated by anti-CD3 antibodies. Remarkably, we also show the recruitment of the ubiquitin ligase Cbl-b and a significant ubiquitination at the immunological synapse in antigen-stimulated T-cells. Our results identify the immunological synapse as the cellular area where TCR-induced protein ubiquitination occurs. They imply that the synapse is a specialized site where the activation process is not only triggered, but also controlled via ubiquitination of signaling actors. PMID- 15790511 TI - Significance of the detection of HIV-1 gag- and/or pol-CD8/A2 T-lymphocytes in HIV-patients. AB - Cytotoxic T-lymphocytes (CTL) play an important role in the immune system's defense against human immunodeficiency virus (HIV) infection. The functional status of CTL closely relates to the progression of HIV disease. We have validated the characteristics of the assay for HIV-1 gag- and pol-specific CD8/HLA-A2 T-cells from peripheral blood by flow cytometry. Sixty-nine healthy individuals and 38 HIV-patients with HLA-A2 antigen-positive subjects were included in the study. Neither HIV-1 gag- nor pol-specific-CD8/HLA-A2 T-cells were determined in these healthy subjects. HIV-1 gag- and pol-specific-CD8/HLA-A2 T-cells could be detected in HIV-patients. The frequency of specific CTL was 58% (22/38) in the patient group. There was a significantly inverse correlation (p < 0.05) between HIV-1 gag- and pol-specific-CD8/HLA-A2 T-cells and HIV plasma viremia in the patients. CONCLUSION: The HIV-1 gag- or pol-specific-CD8/HLA-A2 T cells assay is sensitive and specific, being able to detect at the single T-cell level. This assay may provide a versatile tool for structured HIV treatment and for monitoring vaccination efficacy. PMID- 15790510 TI - Mega-dose Vitamin C modulates T cell functions in Balb/c mice only when administered during T cell activation. AB - Previously we reported that a mega-dose of Vitamin C enhanced the initial stage of delayed-type hypersensitivity reaction in Balb/c mice. In this study its effects were further evaluated as follows. Mice were administered Vitamin C intraperitoneally at 0.625 mg/day or at 5mg/day for variable days before, during, or after being sensitized with DNFB. T cells were isolated in each group and examined. When stimulated antigen-specifically or non-specifically in vitro, mice showed elevated thymidine uptake and a shift of cytokine secretion profiles toward Th1, i.e., elevated levels IL-2, TNF-alpha, and IFN-gamma, and lowered level of the Th2 cytokine IL-4, only when Vitamin C was administered during sensitization. T cells from those mice administered Vitamin C before sensitization or after challenge showed the same T cell properties as those from PBS-treated mice. Mice were also given 0.625 mg/day of Vitamin C during primary and/or secondary immunizations with KLH and secondary specific antibody titers in sera were measured. The total specific antibody titer was lowered in Vitamin C treated animals whenever treatments were administered, and this was entirely attributed to decreased levels of IgG1 and IgE antibodies. Based on these results, we suggest that an exogenously administered mega-dose of Vitamin C shifts immunity in Balb/c mouse toward Th1 and that these affects occur only when Vitamin C is administered during T cell activation. PMID- 15790512 TI - Toll-like receptor 9 is expressed on follicle-associated epithelia containing M cells in swine Peyer's patches. AB - The precise distribution and expression of Toll-like receptor (TLR) 9 in gut associated lymphoid tissues (GALTs) has not been elucidated. In this study, we investigated the expression pattern of TLR9 in adult and neonatal swine GALTs by real-time quantitative PCR, western blot, confocal laser microscopy and flow cytometric analysis. The swine TLR9 gene was preferentially expressed in adult Peyer's patches (Pps) and mesenteric lymph nodes (MLNs), which contained approximately three times higher TLR9 than the spleen. Other tissues exhibited only weak expression of TLR9. In neonatal swine, elevated expression of TLR9 was detected only in MLNs. We firstly showed that highly expressive (TLR9(+)) cells were formed in Pps and MLNs. In addition, TLR9(+) cells were present not only in immune cells such as dendritic cells and B cells but also in follicle-associated epithelia (FAE) including membranous cells (M cells) in Pps. These results suggest that Pps and MLNs provide the host defense with the ability to respond to a variety of bioactive oligonucleotides (ODNs) from bacteria at a conductive site of initial immune responses. PMID- 15790513 TI - Requirement for ERK activation in sinomenine-induced apoptosis of macrophages. AB - Sinomenine (SN), an immunnosuppressive compound derived from the Chinese medicinal plant Sinomenium acutum, has been used to treat autoimmune diseases effectively. Previous studies show SN can inhibit lymphocytes proliferation and macrophage production of pro-inflammatory factors. However, little is known about the mechanisms by which SN inhibits macrophage functions. In this study, we demonstrated that SN could inhibit the proliferation of murine macrophages RAW264.7 by inducing apoptosis in a dose- and time-dependent manner. We found activation of extracellular signal-regulated protein kinase (ERK) in SN-treated macrophages, and requirement for ERK activation in SN-induced apoptosis of macrophages. Contemporarily, the expression of p27/KIP1, proapoptotic factor Bax increased, and expression of Bcl-2 decreased, which might cooperate to induce apoptosis. Inhibiting ERK activation reduced the increased expression of p27 and Bax, but had no effect on the decreased expression of Bcl-2, suggesting the involvement of ERK activation in the SN-induced increased expression of p27 and Bax. These results demonstrated that SN could induce apoptosis of macrophages through activation of ERK, and ERK activation might partially involve in the increased expression of p27 and Bax in apoptotic macrophages. Therefore, induction of macrophage apoptosis through ERK activation may be one of mechanisms by which SN exhibits its immunosuppressive function. PMID- 15790514 TI - Tumor necrosis factor-alpha promoter polymorphism in Mexican patients with Chagas' disease. AB - The aim of this study was to investigate whether the promoter's polymorphisms at the TNF alpha (TNF-alpha) gene were associated with the genetic susceptibility to Chagas' disease. We analyzed the TNF-alpha (positions -308, and -238) polymorphisms in a sample of 54 serologically positive chagasic individuals and in 169 healthy controls. The patients were divided according to clinical characteristics as asymptomatics (n = 27), and chronic chagasic cardiopathy (CCC) patients (n = 27). The whole group of patients showed increased frequencies of 308 T2 (A) allele when compared to healthy controls (pC = 0.008, OR = 3.03). When the analysis was carried out separately in asymptomatic and CCC patients, increased frequencies of T2 (A) allele and T1T2 (AG) genotype in the group of patients with CCC were found when compared to asymptomatic individuals (pC = 0.0002 and pC = 0.003, respectively) and healthy controls (pC = 4 x 10(-7), OR = 7.02, and pC = 0.0006, OR = 5.29, respectively). The present study demonstrates that Chagas' disease is associated with TNF-alpha polymorphisms in the Mexican population. The TNF-308 T2 allele could be directly involved in the genetic susceptibility to the chronic phase of the disease. PMID- 15790515 TI - Ligation of RARgamma inhibits proliferation of phytohaemagglutinin-stimulated T cells via down-regulating JAK3 protein levels. AB - The mechanisms whereby Vitamin A regulates the immune system are poorly understood. We have shown previously that retinoic acids, the Vitamin A derivatives, promote both apoptosis of neglected thymocytes and the activation induced cell death of peripheral T-cells via ligating the nuclear retinoid receptor (RAR) gamma. In the present study, we found that human peripheral T cells express RARalpha and gamma, but not RARbeta. Increasing concentrations of 9 cis RA inhibited phytohaemagglutinin (PHA)-induced proliferation of T-cells, an effect that could be mimicked only by addition of RARgamma agonists and could be inhibited by an RARgamma antagonist. Interleukin-2 (IL-2) produced is known to mediate PHA-induced proliferation of T lymphocytes. Ligation of RARgamma did not affect the PHA-induced high affinity IL-2 receptor expression, slightly reduced the PHA-induced IL-2 production, but interfered with the IL-2-mediated signal transduction resulting in inhibition of PHA-induced phosphorylation of retinoblastoma protein and of up-regulation of Bcl-2. Janus kinases JAK1 and JAK3 play a determinant role in IL-2-dependent signal transduction. Ligation of RARgamma did not affect the levels of JAK1, but prevented IL-2-induced expression of JAK3 resulting in inhibition of PHA-induced phosphorylation of Stat5 molecules. Our data suggest that the previously observed toxic effect of high concentrations of retinoids on the immune system might be mediated via formation of 9-cis RA, which via ligation of RARgamma not only induces cell death in immature thymocytes, but inhibits proliferation of T-cells as well. PMID- 15790516 TI - IFN-gamma primes macrophages for enhanced TNF-alpha expression in response to stimulatory and non-stimulatory amounts of microparticulate beta-glucan. AB - beta-(1-->3)-D-Glucan is an integral cell wall component of a variety of fungi, plants, and bacteria. Like the prototypic inflammatory mediator lipopolysaccharide (LPS), some beta-(1--> 3)-D-glucan-containing preparations have been shown to induce the production of proinflammatory cytokines by macrophages. In the present study, we have tested a new microparticulate form of beta-(1--> 3)-D-glucan (MG) from Saccharomyces cerevisiae for its ability to induce proinflammatory cytokine secretion in mouse peritoneal macrophages in vitro, and we have examined the effect of IFN-gamma. MG was rapidly phagocytized by peritoneal macrophages, and these MG-treated macrophages upregulated TNF alpha, IL-6, and IL-1beta mRNAs and secreted these proinflammatory cytokines. IFN gamma treatment alone did not induce unstimulated macrophages to produce TNF alpha. However, a 4 h IFN-gamma pretreatment augmented TNF-alpha secretion by peritoneal macrophages subsequently treated with an optimally stimulatory dose of MG. IFN-gamma pretreatment for 2 h followed by thorough washing and a further 2 h incubation without IFN-gamma still resulted in enhanced TNF-alpha production in response to MG, suggesting that IFN-gamma can prime macrophages for a subsequent proinflammatory response. Most interestingly, we found that IFN-gamma pretreatment of peritoneal macrophages enhanced the TNF-alpha response to amounts of MG that were poorly stimulatory or non-stimulatory in the absence of IFN-gamma priming. These data suggest that a synergy between IFN-gamma and beta-glucan may have evolved to lower the threshold of sensitivity of the innate immune response to fungal pathogens. PMID- 15790518 TI - Human immunodeficiency virus gp120 downregulates CD1d cell surface expression. AB - CD1d is an MHC class I-like surface molecule that presents endogenous glycoplipid antigens. The effect of HIV infection on CD1d surface expression has not yet been reported. FACS analysis revealed significantly lower levels of CD1d on CD14(+) monocytes from HIV-infected subjects compared to HIV-infected subjects on HAART and healthy controls. CD1d expression correlated inversely with viral load in infected individuals. CD1d surface expression on human cell lines was downregulated after infection with M-tropic HIV, T-tropic HIV, or after exposure to HIV gp120 in vitro. These data suggest that CD1d-mediated responses are altered during HIV infection and may thus contribute to the global immunodeficiency seen in these patients. PMID- 15790517 TI - Potent activation of antigen-specific T cells by antigen-loaded nanospheres. AB - Polystyrene nanospheres (NS) were found to be efficiently taken up by murine antigen-presenting cells (APC), especially bone marrow-derived dendritic cells (DC), in vitro and in vivo. The efficiency of NS uptake was not affected by the maturation state of DC. Both immature and mature DC had similar ability to take up NS in a dose- and time-dependent manner. Uptake and intracellular localization of NS was clearly demonstrated by confocal laser microscopy, using NS with fluorescence. DC could efficiently take up ovalbumin (OVA), when loaded on the surface of NS (OVA-NS). Consequently, OVA-NS-pulsed DC activated antigen-specific interferon (IFN)-gamma-producing T cells much more strongly than OVA-pulsed DC in vitro. These results suggest that NS can be used as an efficient antigen delivery system to DC for a variety of vaccines, such as an anti-human immunodeficiency virus type 1 vaccine. PMID- 15790519 TI - The synergistic effect of phytohemagglutinin and interferon-gamma on the expression of tumor necrosis factor-alpha from RAW 264.7 cells. AB - Tumor necrosis factor-alpha (TNF-alpha) is a major cytokine of host immune reaction by foreign agents. Phytohemagglutinin (PHA) is a dynamic contributor to mitogenic stimulation and augmentation of host immune defense. Interferon-gamma (IFN-gamma) induces induction of cytokines in macrophages and lymphocytes. The aim of this study was to examine the synergistic effects of PHA plus low dose IFN gamma on TNF-alpha mRNA production, cytosolic levels, and secretion in RAW 264.7 cells. The cells were stimulated with PHA or IFN-gamma using various concentrations for various times. The effects of PHA on TNF-alpha expression appeared in dose- and time-dependent manners. The maximum doses of PHA and IFN gamma to produce them were 300 microg/ml PHA and 10 ng/ml IFN-gamma. The optimum time of PHA for the TNF-alpha mRNA production and release were 6 and 7 h after stimulation, respectively, whereas the time of IFN-gamma on them was achieved at 3 and 8 h. Although the TNF-alpha mRNA production, cytosolic levels, and secretion from the cells were slightly detected under 10 microg/ml PHA and 1 ng/ml IFN-gamma, the combination of PHA (10 microg/ml) and IFN-gamma (1 ng/ml) greatly increased them, indicating the synergistic effect of PHA plus low dose IFN-gamma on TNF-alpha expression. PMID- 15790520 TI - Induction of CD69 antigen expression in peripheral blood mononuclear cells on exposure to silica, but not by asbestos/chrysotile-A. AB - While cases of silicosis are often complicated by various autoimmune disorders, patients with asbestosis develop malignant tumors such as lung cancer and malignant mesothelioma. These differences may derive from different biological effects, particularly on immunological cells, of silica and asbestos. To find differences between silica and asbestos, the early activation antigen, CD69, on T cells was examined because dysregulated and continuous activation of T cells may promote the survival of self-recognizing T cells. After cultivation of peripheral blood mononuclear cells with or without silica or chrysotile-A, an asbestos, only silica induced CD69 expression on the lymphocytes. This induction of CD69 expression was mediated by protein kinase C activation. In addition, cell-cell contact mediated by HLA-DR was more important than soluble factors secreted from silica-phagocytosed cells such as IL-1beta, IL-6, and IL-8, even though IL-6 and IL-8 were produced during the culture of PBMCs with silica and chrysotile-A. It should be examined how these activated, CD69-expressing lymphocytes affect other immune systems as well as alter themselves in terms of cytokine production and cell-cell interaction, leading to autoimmune disorders in silicosis patients. PMID- 15790521 TI - Heat shock up-regulates TLR9 expression in human B cells through activation of ERK and NF-kappaB signal pathways. AB - Toll-like receptors (TLRs) play a critical role in innate immunity and TLR9 is essential for CpG ODN signaling. As "dangerous signal", heat shock may regulate immune response. However, little is known about TLRs expression and signaling after heat shock. In this study, we investigated regulation of TLR9 expression and function in human B cell line RPMI8226 by heat shock. We demonstrated that TLR9 expression was up-regulated remarkably following heat shock. Coincidently, CpG ODN stimulation significantly increased IL-6 production and up-regulated expressions of MHC I, MHC II and CD86 by heat-shocked B cells. Heat shock activated ERK and NF-kappaB signal pathways, and pretreatment of B cells with specific inhibitors of ERK or NF-kappaB signal pathways inhibited heat shock induced up-regulation of TLR9 expression. These results demonstrated that heat shock promotes TLR9 expression and signaling through activation of ERK and NF kappaB signal pathways in B cells, suggesting that heat shock might modulate host immune response by regulating TLR expression. PMID- 15790522 TI - Effect of a novel interleukin-5 receptor antagonist, YM-90709, on antigen-induced eosinophil infiltration into the airway of BDF1 mice. AB - A newly synthesized compound, YM-90709, 2,3-dimethoxy-6,6-dimethyl-5,6 dihydrobenzo[7,8]indolizino[2,3-b]quinoxaline, was previously reported to specifically inhibit the binding of interleukin-5 (IL-5) to its receptor (R) on human eosinophils. In this study, the intravenous injection of YM-90709 inhibited antigen-induced infiltration of eosinophils into the bronchoalveolar lavage fluid (BALF) of BDF1 mice, with an ED(50) value of 0.050mg/kg. Anti-murine IL-5 monoclonal antibody (mAb) also inhibited the infiltration of eosinophils with an ED(50) value of 0.035mg/kg. These results indicate that YM-90709, which is a novel IL-5R antagonist, inhibits antigen-induced eosinophil recruitment into the airway, the same as anti-IL-5 mAb does. PMID- 15790523 TI - The role of soluble interleukin-6 receptor in inflammatory diseases. PMID- 15790525 TI - Pathogenic effects of alpha-synuclein aggregation. AB - Biochemical and genetic evidence point towards alpha-synuclein aggregation as having a pivotal role in the onset and progression of several neurodegenerative disorders, including Parkinson's disease, multiple system atrophy and Lewy body dementia. We review recent data on how alpha-synuclein aggregates may impact on cellular homeostatic mechanisms including cellular transport and degradation and transcriptional regulation. alpha-Synuclein aggregates can exist as several molecular species and their different features are discussed in the context of the methodologies used for their study and the many chemical and physical factors that influence their formation. PMID- 15790526 TI - Synuclein, dopamine and oxidative stress: co-conspirators in Parkinson's disease? AB - The etiology of Parkinson's disease (PD) is presently unknown. The unifying hallmark of disease is depletion of dopamine and loss of nigrostriatal dopamine neurons. Familial and sporadic forms of the disease are described. The familial mutations occur within alpha-synuclein and molecules involved in protein degradation and mitochondrial function. Sporadic PD is thought to involve the interplay of genetic and environmental factors. Despite disparate initiating triggers, a convergent pathobiologic model for this common neurodegenerative disease has been proposed. Likely players have emerged that may form the basis for this common pathway model of disease. In this review, we examine the role of three most implicated PD pathogenic conspirators: synuclein, dopamine and oxidative stress. PMID- 15790527 TI - Parkinson's disease: in vivo assessment of disease progression using positron emission tomography. AB - Over the past two decades, positron emission tomography (PET) has provided valuable insights into the mechanisms of nigrostriatal degeneration in Parkinson's disease (PD). Furthermore, it allows the in vivo assessment of disease progression and the evaluation of treatment interventions. In this review, we shall discuss some of the issues and concerns that arise with the use of PET as a surrogate marker of disease progression in Parkinson's disease. PMID- 15790528 TI - Stem cells may reshape the prospect of Parkinson's disease therapy. AB - The concept of cell replacement to compensate for cell loss and restore functionality has entered several disease entities including neurodegenerative disorders. Recent clinical studies have shown that transplantation of fetal dopaminergic (DA) cells into the brain of Parkinson's disease (PD) patients can reduce disease-associated motor deficits. However, the use of fetal tissue is associated with practical and ethical problems including low efficiency, variability in the clinical outcome and controversy regarding the use of fetuses as donor. An alternative cell resource could be embryonic stem (ES) cells, which can be cultivated in unlimited amounts and which have the potential to differentiate into mature DA cells. Several differentiation protocols have been developed, and some progress has been made in understanding the mechanisms underlying DA specification in ES cell development, but the "holy grail" in this paradigm, which is the production of sufficient amounts of the "right" therapeutic DA cell, has not yet been accomplished. To achieve this goal, several criteria on the transplanted DA cells need to be fulfilled, mainly addressing cell survival, accurate integration in the brain circuitry, normal function, no tumor formation, and no immunogenicity. Here, we summarize the current state of ES cell-derived DA neurogenesis and discuss the aspects involved in generating an optimal cell source for cell replacement in PD. PMID- 15790529 TI - Redox cycling of the herbicide paraquat in microglial cultures. AB - Mechanisms involved in paraquat neurotoxicity that selectively target nigrostriatal dopaminergic neurons remain relatively unknown. In this study, we tested the hypotheses that paraquat exposure leads to the production of reactive oxygen species (ROS) through a process of redox cycling and that microglia represent an important site for the initiation of redox cycling reactions. Addition of paraquat to N9 microglial cultures resulted in a dose- and time dependent release of superoxide radicals. Other agents that share with paraquat the property of redox cycling, i.e., benzyl viologen and diquat, also induced a marked production of superoxide radicals by microglia. The ability of paraquat, benzyl viologen, and diquat to induce superoxide release was correlated to their one-electron reduction potentials and thus their tendency to redox cycle. Nitric oxide synthase and NADPH oxidase were identified as enzymatic sources of electrons that triggered paraquat redox cycling by microglia. Taken together, these data provide evidence in favor of a new mechanism by which microglia could play a role in oxidative injury during neurodegenerative processes. Microglial NOS and NADPH oxidase could promote the generation of ROS via the redox cycling of paraquat-like toxicants. PMID- 15790530 TI - The MPTP model of Parkinson's disease. AB - The biochemical and cellular changes that occur following administration of 1 methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) are remarkably similar to that seen in idiopathic Parkinson's disease (PD). In this review, we detail the molecular activities of this compound from peripheral intoxication through its various biotransformations. In addition, we detail the interplay that occurs between the different cellular compartments (neurons and glia) that eventually consort to kill substantia nigra pars compacta (SNpc) neurons. PMID- 15790531 TI - Metallothionein-mediated neuroprotection in genetically engineered mouse models of Parkinson's disease. AB - Parkinson's disease is characterized by a progressive loss of dopaminergic neurons in the substantia nigra zona compacta, and in other sub-cortical nuclei associated with a widespread occurrence of Lewy bodies. The cause of cell death in Parkinson's disease is still poorly understood, but a defect in mitochondrial oxidative phosphorylation and enhanced oxidative and nitrative stresses have been proposed. We have studied control(wt) (C57B1/6), metallothionein transgenic (MTtrans), metallothionein double gene knock (MTdko), alpha-synuclein knock out (alpha-syn(ko)), alpha-synuclein-metallothionein triple knock out (alpha-syn MTtko), weaver mutant (wv/wv) mice, and Ames dwarf mice to examine the role of peroxynitrite in the etiopathogenesis of Parkinson's disease and aging. Although MTdko mice were genetically susceptible to 1, methyl, 4-phenyl, 1,2,3,6 tetrahydropyridine (MPTP) Parkinsonism, they did not exhibit any overt clinical symptoms of neurodegeneration and gross neuropathological changes as observed in wv/wv mice. Progressive neurodegenerative changes were associated with typical Parkinsonism in wv/wv mice. Neurodegenerative changes in wv/wv mice were observed primarily in the striatum, hippocampus and cerebellum. Various hallmarks of apoptosis including caspase-3, TNFalpha, NFkappaB, metallothioneins (MT-1, 2) and complex-1 nitration were increased; whereas glutathione, complex-1, ATP, and Ser(40)-phosphorylation of tyrosine hydroxylase, and striatal 18F-DOPA uptake were reduced in wv/wv mice as compared to other experimental genotypes. Striatal neurons of wv/wv mice exhibited age-dependent increase in dense cored intra neuronal inclusions, cellular aggregation, proto-oncogenes (c-fos, c-jun, caspase 3, and GAPDH) induction, inter-nucleosomal DNA fragmentation, and neuro apoptosis. MTtrans and alpha-Syn(ko) mice were genetically resistant to MPTP Parkinsonism and Ames dwarf mice possessed significantly higher concentrations of striatal coenzyme Q10 and metallothioneins (MT 1, 2) and lived almost 2.5 times longer as compared to control(wt) mice. A potent peroxynitrite ion generator, 3 morpholinosydnonimine (SIN-1)-induced apoptosis was significantly attenuated in MTtrans fetal stem cells. These data are interpreted to suggest that peroxynitrite ions are involved in the etiopathogenesis of Parkinson's disease, and metallothionein-mediated coenzyme Q10 synthesis may provide neuroprotection. PMID- 15790532 TI - Effects of DJ-1 mutations and polymorphisms on protein stability and subcellular localization. AB - Mutations in the DJ-1 gene are associated with recessive, early onset Parkinson's disease (PD). We reported previously that one of the point mutations, L166P, destabilizes the protein and thus produces an effective knockout of the gene. Here, we have expanded this analysis to include a series of mutations and polymorphisms identified throughout the gene. The M26I point mutation was also unstable, although the effect was not as dramatic as with L166P. Protein levels were rescued in part, but not completely, by proteasome inhibition. Other variants, such as R98Q, were generally stable. We noted that M26I and L166P are both in helical regions near the dimer interface. However, M26I retains the ability to dimerize. We also examined the subcellular localization of DJ-1 and found that most mutations were similar to the wild-type (wt) protein in that a few cells showed mitochondrial staining. However, in all cases, the proportion of cells with mitochondrial DJ-1 staining was increased in oxidative conditions, suggesting that oxidation promotes the mitochondrial localization of DJ-1. PMID- 15790533 TI - Effects of nitration on the structure and aggregation of alpha-synuclein. AB - Substantial evidence suggests that the aggregation of the presynaptic protein alpha-synuclein is a key step in the etiology of Parkinson's disease (PD). Although the molecular mechanisms underlying alpha-synuclein aggregation remain unknown, oxidative stress has been implicated in the pathogenesis of PD. Here, we report the effects of tyrosine nitration on the propensity of human recombinant alpha-synuclein to fibrillate in vitro. The properties of nitrated alpha synuclein were investigated using a variety of biophysical and biochemical techniques, which revealed that nitration led to formation of a partially folded conformation with increased secondary structure relative to the intrinsically disordered structure of the monomer, and to oligomerization at neutral pH. The degree of self-association was concentration-dependent, but at 1 mg/mL, nitrated alpha-synuclein was predominantly an octamer. At low pH, small-angle X-ray scattering data indicated that the nitrated protein was monomeric. alpha Synuclein fibrillation at neutral pH was completely inhibited by nitrotyrosination and is attributed to the formation of stable soluble oligomers. The presence of heparin or metals did not overcome the inhibition; however, the inhibitory effect was eliminated at low pH. The addition of nitrated alpha synuclein inhibited fibrillation of non-modified alpha-synuclein at neutral pH. Potential implications of these findings to the etiology of Parkinson's disease are discussed. PMID- 15790534 TI - Absence of inclusion body formation in the MPTP mouse model of Parkinson's disease. AB - Formation of alpha-synuclein aggregation and Lewy bodies (LBs) are hallmarks of Parkinson's disease (PD) and other related diseases. The dopaminergic neurotoxin, MPTP, replicates many of the pathological signs and motoric features of PD in primates and rodents by selective destruction of dopamine (DA) neurons of the substantia nigra. In this study, groups of adult wild-type C57BL6 mice were treated with MPTP either acutely (20 mg/kg, every 2 h x 4 for 1 day), semi chronically (30 mg/kg/day for 5 days), or chronically (25 mg/kg MPTP with 250 mg/kg probenecid 2 times/week for 5 weeks). Mice brains were collected and processed at various time points for immunohistochemistry and HPLC assays. Our data showed that although there is a significant decrease in DA content and its metabolites and tyrosine hydroxylase immunoreactivity, there is no inclusion body formation following the various MPTP treatment regimens. PMID- 15790535 TI - Rotenone induces oxidative stress and dopaminergic neuron damage in organotypic substantia nigra cultures. AB - Rotenone, a pesticide and complex I inhibitor, causes nigrostriatal degeneration similar to Parkinson disease pathology in a chronic, systemic, in vivo rodent model [M. Alam, W.J. Schmidt, Rotenone destroys dopaminergic neurons and induces parkinsonian symptoms in rats, Behav. Brain Res. 136 (2002) 317-324; R. Betarbet, T.B. Sherer, G. MacKenzie, M. Garcia-Osuna, A.V. Panov, J.T. Greenamyre, Chronic systemic pesticide exposure reproduces features of Parkinson's disease, Nat. Neurosci. 3 (2000) 1301-1306; S.M. Fleming, C. Zhu, P.O. Fernagut, A. Mehta, C.D. DiCarlo, R.L. Seaman, M.F. Chesselet, Behavioral and immunohistochemical effects of chronic intravenous and subcutaneous infusions of varying doses of rotenone, Exp. Neurol. 187 (2004) 418-429; T.B. Sherer, J.H. Kim, R. Betarbet, J.T. Greenamyre, Subcutaneous rotenone exposure causes highly selective dopaminergic degeneration and alpha-synuclein aggregation, Exp. Neurol. 179 (2003) 9-16.]. To better investigate the role of mitochondria and complex I inhibition in chronic, progressive neurodegenerative disease, we developed methods for long-term culture of rodent postnatal midbrain organotypic slices. Chronic complex I inhibition over weeks by low dose (10-50 nM) rotenone in this system lead to dose- and time dependent destruction of substantia nigra pars compacta neuron processes, morphologic changes, some neuronal loss, and decreased tyrosine hydroxylase (TH) protein levels. Chronic complex I inhibition also caused oxidative damage to proteins, measured by protein carbonyl levels. This oxidative damage was blocked by the antioxidant alpha-tocopherol (vitamin E). At the same time, alpha tocopherol also blocked rotenone-induced reductions in TH protein and TH immunohistochemical changes. Thus, oxidative damage is a primary mechanism of mitochondrial toxicity in intact dopaminergic neurons. The organotypic culture system allows close study of this and other interacting mechanisms over a prolonged time period in mature dopaminergic neurons with intact processes, surrounding glia, and synaptic connections. PMID- 15790536 TI - Quantitative proteomic analysis of mitochondrial proteins: relevance to Lewy body formation and Parkinson's disease. AB - The mechanisms underlying Parkinson's disease (PD) and Lewy body (LB) formation, a pathological hallmark of PD, are incompletely understood; however, mitochondrial dysfunction is likely to be at least partially responsible. To study the processes that might be related to nigral neurodegeneration and LB formation, we employed nonbiased quantitative proteomics with isotope-coded affinity tag (ICAT) to compare the mitochondrial protein profiles in the substantia nigra (SN) between controls and mice treated chronically with 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a potent mitochondrial toxicant, and an adjuvant, probenecid (prob), for 5 weeks, which produced selective nigrostriatal neurodegeneration with formation of LB-like cytoplasmic inclusions in the remaining nigral neurons. This method identified a total of more than 300 proteins; of these proteins, more than 100 displayed significant changes in relative abundance in the MPTP/prob-treated mice compared to the controls. We validated one of these proteins, DJ-1, whose mutation has been implicated in familial PD, with Western blot analysis, followed by immunohistochemical studies of its distribution in the SN in relation to cytoplasmic inclusions in mice, as well as in classical LBs in PD patients. The results demonstrated that DJ-1 was not only colocalized with alpha-synuclein in dopaminergic neurons but also to cytoplasmic inclusions in mice treated with MPTP/prob. In addition, DJ-1 was present in the halo but not in the core of classical LBs in patients with PD. Our findings suggested that DJ-1 might play an important role in mitochondrial dysfunction, as well as LB formation in PD. PMID- 15790537 TI - Dopaminergic innervation of forebrain by ventral mesencephalon in organotypic slice co-cultures: effects of GDNF. AB - Numerous studies have verified the ability of glial cell line-derived neurotrophic factor (GDNF) to protect or rescue neurons in models of Parkinson's disease. However, the role of GDNF in the development of dopaminergic (DA) neurons remains unclear. We investigated the hypothesis that GDNF is a target protein for the DA neurons of the mesencephalon forming the nigrostriatal pathway in an in vitro rat model. Organotypic slice cultures were prepared from tissue isolated from postnatal rat pups including but not limited to the substantia nigra (SN), striatum, and cerebral cortex. These cultures were maintained for up to 100 days in vitro. In the absence of exogenous GDNF, DA neurons from the SN grew into the striatum but not the cerebral cortex or hippocampus as determined by immunostaining for tyrosine hydroxylase. The addition of exogenous GDNF increased the survival of DA neurons and also enhanced the number of dopaminergic processes innervating the striatum. GDNF also induced DA innervation of the cerebral cortex but not hippocampus. In conclusion, our studies indicate that the normal pattern of innervation by DA neurons of the mesencephalon can be recapitulated with organotypic co-cultures and that this pattern can be altered by GDNF. PMID- 15790539 TI - Delayed gene therapy of glial cell line-derived neurotrophic factor is efficacious in a rat model of Parkinson's disease. AB - Gene transfer of glial cell line-derived neurotrophic factor (GDNF) in rodent models of Parkinson's disease (PD) has been shown to protect against neurodegeneration either prior to or immediately after neurotoxin-induced lesions; however, the nigrostriatal pathway was largely intact when gene delivery was completed in these models, which may not accurately reflect the clinical situation encountered with Parkinson's patients. In this study, replication incompetent adenoviral vectors encoding the rat GDNF gene were administered into the striatum 4 weeks following 6-hydroxydopamine (6-OHDA) injection in the unilateral striatum, more closely resembling fully developed PD. Apomorphine induced rotational behavior testing was performed every week following 6-OHDA injection. At the 10th week after gene transfer, the striatal dopamine concentrations were measured by HPLC with an electrochemical detector and the number of tyrosine hydroxylase (TH)-positive dopamine neurons in the substantia nigra (SN) was determined by immunohistochemistry. Injection of 6-OHDA into the striatum produced stable increases in rotation, which reached a plateau between 4 and 5 weeks post-injection. The number of TH-positive neuron in the SN and dopamine levels in the striatum was significantly lower in the 6-OHDA group compared to the normal group. Gene transfer of GDNF, but not beta-galactosidase, significantly increased the number of TH-positive neurons and dopamine levels, with a subsequent behavioral recovery between 5 and 10 weeks following GDNF transduction. These findings demonstrate that adenovirus-mediated gene transfer of GDNF is efficacious even in the late stages of 6-OHDA-induced PD rats. They also provide further evidence on the effectiveness of GDNF-based gene therapy for experimental Parkinson's disease. PMID- 15790538 TI - Endogenous brain-derived neurotrophic factor protects dopaminergic nigral neurons against transneuronal degeneration induced by striatal excitotoxic injury. AB - Injury to the central nervous system causes atrophy or death of connecting neurons and can modify the expression of neurotrophic factors. We observed transneuronal upregulation of brain-derived neurotrophic factor (BDNF) expression in the rat ipsilateral substantia nigra pars compacta after a striatal lesion induced by kainate. This effect is developmentally regulated because the enhancement of nigral BDNF expression was only observed when striatal lesion was performed on postnatal day (P) 15 and in adulthood, but not at P7. Interestingly, the lack of regulation of BDNF was coincident with the transynaptic degeneration of nigral neurons after striatal excitotoxic injury. Hence, the number of tyrosine hydroxylase-positive neurons in the substantia nigra pars compacta decreased when the lesion was performed at P7, but not at P15 or at P30. The analysis of the functional significance of this BDNF upregulation was done using trkB-IgG fusion proteins. After striatal injury, blockade of endogenous BDNF by trkB fusion proteins induced an atrophy of the dopaminergic neurons of the pars compacta. The injection of trkB-IgG fusion proteins did not modify the effects of kainate in the substantia nigra pars reticulata. Thus, our results show that BDNF exerts an autocrine/paracrine protective effect selectively on dopaminergic neurons against the loss of trophic support from the target striatum. PMID- 15790540 TI - Valproate pretreatment protects dopaminergic neurons from LPS-induced neurotoxicity in rat primary midbrain cultures: role of microglia. AB - Parkinson's disease is a neurodegenerative disorder characterized by progressive degeneration of dopaminergic (DA) neurons in the substantia nigra. Accumulating evidence supports the notion that neuroinflammation is involved in the pathogenesis of this disease. Valproate (VPA) has long been used for the treatment of seizures and bipolar mood disorder. In vivo and in vitro studies have demonstrated that VPA has neuroprotective and neurotrophic actions. In this study, using primary neuron-glia cultures from rat midbrain, we demonstrated that VPA is a potent neuroprotective agent against lipopolysaccharide (LPS)-induced neurotoxicity. Results showed that pretreatment with 0.6 mM VPA for 48 h robustly attenuated LPS-induced degeneration of dopaminergic neurons as determined by [(3)H] dopamine uptake and counting of the number of TH-ir neurons. The neuroprotective effect of VPA was concentration-dependent and was mediated, at least in part, through a decrease in levels of pro-inflammatory factors released from activated microglia. Specifically, LPS-induced increase in the release of TNFa, NO, and intracellular reactive oxygen species was markedly reduced in cultures pretreated with VPA. These anti-inflammatory effects of VPA were time and concentration-dependent correlated with a decrease in the number of microglia. Thus, our results demonstrate that protracted VPA pretreatment protects dopaminergic neurons from LPS-induced neurotoxicity through a reduction in levels of released pro-inflammatory factors, and further suggest that these anti-inflammatory effects may be contributed by VPA-induced reduction of microglia cell number. Taken together, our study reinforces the view that VPA may have utility in treating Parkinson's disease. PMID- 15790542 TI - Cancer cell death by programmed necrosis? AB - A recent paper by Zong et al. describes how alkylating agents kill cells by a process they term "programmed necrosis," induced by excessive activation of PARP resulting in degradation of cytosolic NAD(+) and inhibition of glycolysis. We argue that it is not obvious whether chemotherapy in patients can induce sufficient NAD(+) loss to affect glycolysis; that the "programmed" nature of the necrosis requires more evidence; and that there are mechanisms making cancer cells hypersensitive to DNA damage other than their high rate of aerobic glycolysis. PMID- 15790543 TI - Individualized chemotherapy treatment for breast cancer: is it necessary? Is it feasible? AB - Individualized selection of chemotherapy regimens based on the molecular characteristics of the cancer would be desirable. However, it is not currently feasible. The development of clinically useful predictors of response to chemotherapy has proven to be difficult. Recently, novel analytical tools particularly transcriptional profiling, have shown promise as potential predictive tests in several small exploratory studies. However, many of the practical challenges of clinical pharmacogenomics are the same that have plagued prognostic and predictive marker research for decades. Perhaps, the most important challenge is to prospectively design and conduct validation trials that demonstrate clinical utility by showing improved patient outcomes with the use of a proposed new test. PMID- 15790541 TI - Environmental enrichment in adulthood eliminates neuronal death in experimental Parkinsonism. AB - Idiopathic Parkinson's disease (PD) affects 2% of adults over 50 years of age. PD patients demonstrate a progressive loss of dopamine neurons in the substantia nigra pars compacta (SNpc). One model that recapitulates the pathology of PD is the administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Here we show that exposure to an enriched environment (EE) (a combination of exercise, social interactions and learning) or exercise alone during adulthood, totally protects against MPTP-induced Parkinsonism. Furthermore, changes in mRNA expression would suggest that increases in glia-derived neurotrophic factors, coupled with a decrease of dopamine-related transporters (e.g. dopamine transporter, DAT; vesicular monoamine transporter, VMAT2), contribute to the observed neuroprotection of dopamine neurons in the nigrostriatal system following MPTP exposure. This non-pharmacological approach presents significant implications for the prevention and/or treatment of PD. PMID- 15790544 TI - Chemotherapeutic potential of the chemopreventive phytoalexin resveratrol. AB - The last couple of decades have seen a tremendous increase in interest in the biological properties of natural products as a means to identify novel small compounds that could have potential in clinical medicine. To that end, flavonoids and flavonoid-like compounds percolate to the top due to their presence in diet constituents and reported beneficial effects on diverse biological processes and disease conditions. As such, the plant polyphenolic antibiotic resveratrol, found in grapes, nuts and wines, has been the focus of many studies aimed at understanding its full range of health beneficial effects. The interest in this compound stems from the earlier observations describing the therapeutic benefits of roots of the oriental medicinal plant from which resveratrol was first isolated. Being a constituent of grapes and wines, the initial work was focused on linking resveratrol to the beneficial cardiovascular effects of moderate wine intake, however, since its reported cancer chemopreventive activity in a murine model of carcinogenesis, there has been a heightened interest in understanding the anti-cancer activity of resveratrol. As a result, a substantial amount of data strongly suggests that resveratrol could affect the process of carcinogenesis through a variety of different mechanisms in different tumor cell types. However, a couple of recent reports provide evidence to the contrary. This critical review attempts to summarize some of these findings and discuss the clinical potential of this compound or its derivatives in the light of the recent conflicting reports. PMID- 15790546 TI - Apoptosis in infectious mononucleosis: its detection with the Beckman Coulter GenS hematology analyzer. AB - This article describes the potential of the Beckman Coulter GenS hematology analyzer for detecting and quantifying the phenomenon of elevated apoptosis found during the incubation at 37 degrees C of samples containing CD45RO+ reactive T lymphocytes from patients with infectious mononucleosis (IM). Visual detection was carried out with the GenS DF1 scattergram (volume/scatter), and the means and SDs of lymphocyte positional parameters (volume, scatter, and conductivity) were recorded from the instrument research screen before and after samples were incubated in an oven for 4 hours. Thirty-one IM samples were analyzed, and the results obtained after sample incubation showed a mean reduction in lymphocyte volume of 34%, a 3.9% increase in scatter, and a 4% increase in conductivity. A new cluster of cells consistent with apoptotic lymphocytes appeared in the GenS DF1 scattergram. May-Gr-Giemsa staining and extended manual counting of the samples carried out at the same time to validate and quantify the apoptosis of the analyzed samples detected a significant increase in apoptotic cells from 1.2% (range, 0%-3%) to 8.37% (range, 1%-39%) after incubation. The same process was used to evaluate a control group of patients, and the possible interference of platelet aggregation and erythroblasts (nucleated red blood cells) in GenS apoptosis quantification was investigated. The final results showed that the appearance of the apoptotic cluster in the GenS DF1 scattergram occurred in 96.8% of the IM cases, although this phenomenon is apparently nonspecific because similar clusters appear in other pathologies, such as viral infections (hepatitis, cytomegalovirus, and acquired immune deficiency syndrome). Additional quantitative studies of apoptosis kinetics indicated that at least 4 hours of incubation are necessary. This finding has directed an investigation of apoptosis inducing drugs (camptothecin, theophylline) to reduce incubation time and thereby enhance the practical application of automated apoptosis detection in the diagnosis of IM and other diseases in which the phenomenon of apoptosis occurs. PMID- 15790545 TI - Lessons from TRAIL-resistance mechanisms in colorectal cancer cells: paving the road to patient-tailored therapy. AB - Colorectal cancer is one of the leading causes of cancer-related deaths worldwide. Intrinsic, as well as acquired, resistance to chemotherapy remains a major problem in the treatment of this disease. It is, therefore, of great importance to develop new, patient-tailored, treatment strategies for colorectal cancer patients. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) acts through the pro-apoptotic DR4 and DR5 receptors in tumor cells without harming normal cells and will soon be tested in clinical trials as a novel anti cancer agent. However, not all human colon cancer cell lines are sensitive to TRAIL due to intrinsic or acquired TRAIL-resistance. This review discusses the mechanisms and modulation of TRAIL-resistance in colon cancer cells. Cell sensitivity to TRAIL can be affected by TRAIL-receptor expression at the cell membrane, DR4/DR5 ratio and functionality of TRAIL-receptors. Additional intracellular factors leading to TRAIL-resistance affect the caspase 8/c-FLIP ratio, such as loss of caspase 8 and caspase 10 due to mutations or gene methylation, CARP-dependent degradation of active caspase 8 and changes in caspase 8 or c-FLIP expression levels. Further downstream in the TRAIL apoptotic pathway, Bax mutations, or increased expression of IAP family members, in particularly XIAP and survivin, also cause resistance. Chemotherapeutic drugs, NSAIDs, interferon-gamma and proteasome inhibitors can overcome TRAIL-resistance by acting on TRAIL-receptor expression or changing the expression of pro- or anti apoptotic proteins. PMID- 15790547 TI - An ATL cell line with an IgH pseudo-rearranged band pattern by southern blotting: a pitfall of genetic diagnosis. AB - Adult T-cell leukemia (ATL), which is a mature T-cell malignancy that develops from human T-cell leukemia virus type-1 (HTLV-1)-infected T-cells, is diagnosed based on morphologic, immunophenotypic, serologic, and genetic characteristics. In particular, Southern blot hybridization (SBH) and polymerase chain reaction analyses for antigen receptor genes and the retrovirus of HTLV-1 provide a diagnostic hallmark for the clonality of leukemic cells and the causative agent of the disease. We report here a case of an ATL cell line, designated as SO4 cells, established from primary ATL cells presenting with an irrational genetic abnormality of the immunoglobulin heavy chain (IgH)-rearranged band in spite of harboring a clonally rearranged T-cell receptor gene and a clonally integrated provirus of HTLV-1 within their genomic DNA. Moreover, fluorescence in situ hybridization analysis using the IgH (14q32) dual-color break-apart probe revealed 3 pair-signals of colocalizing red and green spots, implying 2 intact and 1 amplified 14q32 regions without translocation, where the region contains the IgH gene locus. Although the exact mechanism remains to be elucidated, some alteration of a portion of the amplified 14q32 region seems to have a role in the false-positive band pattern in the SBH. The SO4 cells, in the hematology laboratory, will provide a lesson about the pitfalls of genetic tests for mature T-cell neoplasms and contribute to the genetic elucidation of leukomogenesis as an in vitro model. PMID- 15790548 TI - Anemia of chronic disease: pathophysiology and laboratory diagnosis. AB - Classic iron deficiency (ID) does not represent a challenge for the laboratory and physicians. The anemia that accompanies infection, inflammation, and cancer, commonly termed anemia of chronic disease (ACD), features apparently normal or increased iron stores. However, 20% of these patients have iron-restricted erythropoiesis (functional ID), an imbalance between the iron requirements of the erythroid marrow and the actual iron supply. Functional ID leads to a reduction in red cell hemoglobiniza-tion, causing hypochromic microcytic anemia. The diagnosis of functional ID in real time is based on measuring the hemoglobin content of reticulocytes. An examination of the biochemical markers of iron metabolism demonstrates weaknesses in the diagnosis of functional ID. We developed a diagnostic plot for the assessment of iron status in ACD and the detection of advancing ID in patients with ID, ACD, and the combined state of functional ID and ACD. The plot indicates the correlation between a marker of the iron supply for erythropoiesis (ie, the ratio of the soluble transferrin receptor value to the logarithm of the ferritin value) and the reticulocyte hemoglobin content and functions as a marker of iron demand. The diagnostic plot shows good selectivity for assessing the iron status of disease-specific anemias such as classic ID, end-stage renal failure, cancer-related anemia, and the anemia of infection and inflammation. The therapeutic implications of the diagnostic plot are to differentiate patients who should be administered oral iron supplements, recombinant human erythropoietin (r-HuEPO), or a combination of r-HuEPO and iron. The response of erythropoiesis to r-HuEPO depends on the iron supply and the proliferation of erythropoiesis. The lack of an increase or a decrease in reticulocyte hemoglobin levels indicates a nonresponder to r-HuEPO or functional ID. PMID- 15790549 TI - Performance evaluation of the application of body fluids on the Sysmex XE-2100 series automated hematology analyzer. AB - Body fluid analysis on the Sysmex XE-2100 series automated hematology analyzer was evaluated at 4 hospitals (Baylor University Medical Center, Dallas, TX; St. John's Mercy, St. Louis, MO; Carle Clinic, Urbana, IL; and ACL Laboratories, West Allis, WI, USA). The total nucleated cell and red blood cell (RBC) counts of 493 samples were obtained with the Sysmex XE-2100 automated hematology analyzer and compared with results obtained by manual chamber counting. Seventy-eight samples were not suitable for evaluation because of the presence of clots, crystals, error messages related to white blood cell (WBC) and RBC parameters, and so on. Pearson correlation coefficients for the WBC parameter were 0.99 for cerebrospinal fluid, 0.95 for serous fluid, 0.99 for synovial fluid, and 0.99 for samples of combined body fluids. Ninety-six samples were used to compare RBC counting methods because these samples had RBC counts greater than 0.01 10 6/mL. The Pearson correlation coefficients for the RBC parameter were 0.96 for cerebrospinal fluid, 0.97 for serous fluid, 0.97 for synovial fluid, and 0.97 for samples of combined body fluids. Carryover, precision, and linearity studies also performed for WBC and RBC counts yielded very good results. PMID- 15790550 TI - CD36: a multiligand molecule. AB - CD36 is a multiligand receptor associated with a broad array of physiological processes and involved in markedly diverse disorders, including atherosclerosis, insulin resistance and diabetes, dyslipidemia, tumor angiogenesis, and host defense against Plasmodium falciparum. CD36 deficiency has proved to be common, particularly in ethnic groups such as African Americans and Asians. CD36 is commonly expressed on blasts in acute monocytic leukemia, megakaryoblastic leukemia, and erythroleukemia. The role of CD36 in sickle cell crises and cerebral malaria is debatable. As a receptor for thrombospondin 1, CD36 plays a role in the regulation of angiogenesis, which may be a therapeutic strategy for controlling the dissemination of malignant neoplasms. The future challenge will be to further understand the mechanisms by which CD36 affects these diverse functions and to design therapeutic strategies that can alter the course of the diseases. PMID- 15790551 TI - Virtual microscopy:applications to hematology. AB - Virtual microscopy is the simulation of microscopy over a computer network. A virtual slide is a giant digital image file of a glass slide that can be displayed, panned, zoomed, and focused in a virtual slide viewer on a computer screen. Virtual slides represent a revolutionary advance over glass slides. They are easy to file, store, retrieve, annotate, and mark and can be preserved indefinitely. Furthermore, they are easy to duplicate and distribute and can be integrated into electronic patient records. Large virtual slides can be readily transmitted to users over a standard broadband connection. With the recent introduction of viewers that can focus virtual slides, virtual microscopy can simulate all the functions of real microscopy. Virtual microscopy has significant advantages over real microscopy in education and in proficiency testing. In education, virtual microscopy enables "anytime, anywhere" learning and has been favorably received by students and teachers. In proficiency surveys, all users view the same image, virtual slides are easy to distribute, and the slides do not deteriorate. Potential applications for hematology proficiency surveys include blood and bone marrow morphology, differential cell counts, cytochemistry and immunocytochemistry, detection of malarial parasites, and other tests. Virtual microscopy enables proficiency surveys of critical clinical parameters, such as the bone marrow blast count, and implementation of "locate and identify" exercises. It is conceivable that with the next generation of technological developments, virtual microscopy can be extended to diagnostic applications. Important goals are to minimize slide file size without loss of relevant detail, to establish diagnostic equivalence, and to automate virtual slide capture with high throughput for integration into laboratory information systems. Key factors that will drive implementation include user-friendliness, cost, data storage requirements, and throughput speed. Implementation may have constructive effects on teaching and learning, the peer-to-peer consultative process, and diagnostic accuracy and performance. PMID- 15790553 TI - The ADVIA 2120 hematology system: flow cytometry-based analysis of blood and body fluids in the routine hematology laboratory. AB - The ADVIA 2120 Hematology System was recently released by Bayer HealthCare, Diagnostics Division, as a bench-top analyzer designed for medium- to large volume laboratories. This flow cytometry-based system uses light scatter, differential white blood cell (WBC) lysis, and myeloperoxidase and oxazine 750 staining to provide a complete blood cell count, a WBC differential, and a reticulocyte count. A cyanide-free method is used to measure hemoglobin colorimetrically. The system is automation ready; in addition to its capability for analyzing peripheral blood specimens, the analyzer is also equipped to analyze cerebrospinal fluid samples. In this article we explain the underlying technology of the ADVIA 2120, provide linearity ranges, method-specific reference ranges, and stability data, and describe novel parameters and applications that are unique to the methodology used by this instrument. Finally, we discuss research applications and future directions, such as the use of this hematology analyzer in the determination of fetal lung maturity. PMID- 15790554 TI - Performance evaluation of the ADVIA 2120 hematology analyzer: an international multicenter clinical trial. AB - Automated cell counters are widely used in modern clinical laboratories to provide reliable, fast, and cost-effective complete blood counts (CBCs), white blood cell differentials, and reticulocyte measurements. In addition, some advanced instruments provide novel parameters, such as the hemoglobin content of reticulocytes or the percentage of hypochromic cells, and are capable of analysis of a variety of body fluids. Bayer recently introduced the ADVIA 2120 system as an automation-ready cell counter for mid- to high-volume testing in the clinical laboratory. This instrument, which builds on the established technology of the ADVIA 120 system, operates with a cyanide-free method for hemoglobin measurement, has a new user interface, and can routinely analyze biological fluid samples in addition to blood. We used 749 samples from 6 worldwide trial sites to evaluate the clinical performance of this new device. Accuracy of the ADVIA 2120 system versus its predecessor model, the ADVIA 120 system, was excellent for all CBC and white cell differential parameters and reticulocyte counts (all correlation coefficients except for basophils >0.9). Correlation of the white cell differential with the standard manual method and within-run precision of the ADVIA 2120 system also was very good. Use of the novel cyanide-free method for hemoglobin measurement had no clinically significant impact on hemoglobin results, even in patients with hemoglobinopathies. We concluded that the ADVIA 2120 system has clinically equivalent performance to the ADVIA 120 system. PMID- 15790555 TI - Effects of the blood-collection tube material and long-term storage on platelet activation parameters on the ADVIA 120/2120 hematology system. AB - The ADVIA 120/2120 Hematology System provides a number of novel parameters of platelet activation. Although the effects of anticoagulant in the blood collection tube, short-term storage, and ambient temperature on these parameters have been documented in the literature, the effects of long-term storage and the material of the blood-draw tube have not been investigated. We therefore compared platelet activation parameter values obtained with the ADVIA 120 for 50 paired blood samples collected into BD Medical Systems' Vacutainer K(3)EDTA glass tubes and Vacutainer K(2)EDTA Plus plastic tubes and analyzed within 4 hours and after 24 hours following venipuncture. We found a statistically significant but clinically irrelevant decrease (less than 1%) in mean platelet component (MPC) concentration in plastic compared with glass for blood samples analyzed within 4 hours of draw. Reanalysis of the samples after 24 hours showed a clinically and statistically significant increase in mean platelet volume and significant decreases in MPC, platelet component distribution width, and mean platelet mass. These storage-induced changes were of equal magnitude for both tube types. We conclude that the use of plastic versus glass blood-draw tubes should have no clinically significant effect on the parameters of platelet activation on the ADVIA 120/2120 system and that the changes in platelet activation parameters continue to be present at 24 hours. PMID- 15790556 TI - Comparative study of blood cell staining with wright-giemsa stain, field stain, and a new modified stain. AB - Routine staining of blood films usually entails staining with Wright-Giemsa stain; however, some modifications of the standard Wright-Giemsa stain have been performed. The field stain or dip method is an example of these modifications. The authors created a new modification of Wright-Giemsa stain and performed a study to compare our modified stain with Wright-Giemsa stain and field stain. Three experts in microscopy were used in performing this comparative study. The average grading score from each stain method from all 3 experts was set as a basic parameter for comparison. We found that the field stain, which was created for field work, had a lower grading score in staining than the other 2 staining methods in many studied aspects (analysis of variance test, P < .05). In addition, we can demonstrate that the 3 experts presented similar decisions on the blood smear evaluation. Conclusively, we found that our modified method can give staining results similar to the standard Wright-Giemsa stain. In addition, our modification can decrease the staining time. PMID- 15790558 TI - Functional analysis of the plant disease resistance gene Pto using DNA shuffling. AB - Pto is a serine/threonine kinase that mediates resistance in tomato to strains of Pseudomonas syringae pv. tomato expressing the (a)virulence proteins AvrPto or AvrPtoB. DNA shuffling was used as a combinatorial in vitro genetic approach to dissect the functional regions of Pto. The Pto gene was shuffled with four of its paralogs from a resistant haplotype to create a library of recombinant products that was screened for interaction with AvrPto in yeast. All interacting clones and a representative sample of noninteracting clones were sequenced, and their ability to signal downstream was tested by the elicitation of a hypersensitive response in an AvrPto-dependent or -independent manner in planta. Eight candidate regions important for binding to AvrPto or for downstream signaling were identified by statistical correlations between individual amino acid positions and phenotype. A subset of the regions had previously been identified as important for recognition, confirming the validity of the shuffling approach. Three novel regions important for Pto function were validated by site-directed mutagenesis. Several chimeras and point mutants exhibited a differential interaction with (a)virulence proteins in the AvrPto and VirPphA family, demonstrating distinct binding requirements for different ligands. Additionally, the identification of chimeras that are both constitutively active as well as capable of binding AvrPto indicates that elicitation of downstream signaling does not involve a conformational change that precludes binding of AvrPto, as previously hypothesized. The correlations between phenotypes and variation generated by DNA shuffling paralleled natural variation observed between orthologs of Pto from Lycopersicon spp. PMID- 15790557 TI - Disease-associated mutations inactivate AMP-lysine hydrolase activity of Aprataxin. AB - Ataxia-oculomotor apraxia syndrome 1 is an early onset cerebellar ataxia that results from loss of function mutations in the APTX gene, encoding Aprataxin, which contains three conserved domains. The forkhead-associated domain of Aprataxin mediates protein-protein interactions with molecules that respond to DNA damage, but the cellular phenotype of the disease does not appear to be consistent with a major loss in DNA damage responses. Disease-associated mutations in Aprataxin target a histidine triad domain that is similar to Hint, a universally conserved AMP-lysine hydrolase, or truncate the protein NH2-terminal to a zinc finger. With novel fluorigenic substrates, we demonstrate that Aprataxin possesses an active-site-dependent AMP-lysine and GMP-lysine hydrolase activity that depends additionally on the zinc finger for protein stability and on the forkhead associated domain for enzymatic activity. Alleles carrying any of eight recessive mutations associated with ataxia and oculomotor apraxia encode proteins with huge losses in protein stability and enzymatic activity, consistent with a null phenotype. The mild presentation allele, APTX-K197Q, associated with ataxia but not oculomotor apraxia, encodes a protein with a mild defect in stability and activity, while enzyme encoded by the atypical presentation allele, APTX-R199H, retained substantial function, consistent with altered and not loss of activity. The data suggest that the essential function of Aprataxin is reversal of nucleotidylylated protein modifications, that all three domains contribute to formation of a stable enzyme, and that the in vitro behavior of cloned APTX alleles can score disease-associated mutations. PMID- 15790559 TI - Peptides derived from the reverse transcriptase of human immunodeficiency virus type 1 as novel inhibitors of the viral integrase. AB - Recent studies have shown that the integrase (IN) of HIV-1 is inhibited in vitro by HIV-1 reverse transcriptase (RT). We further investigated the specific protein sequences of RT that were involved in this inhibition by screening a complete library of RT-derived peptides for their inhibition of IN activities. Two 20 residue peptides, peptide 4286, derived from the RT DNA polymerase domain, and the one designated 4321, from the RT ribonuclease H domain, inhibit the enzymatic activities of IN in vitro. The former peptide inhibits all three IN-associated activities (3'-end processing, strand transfer, and disintegration), whereas the latter one inhibits primarily the first two functions. We showed the importance of the sequences and peptide length for the effective inhibition of IN activities. Binding assays of the peptides to IN (with no DNA substrate present) indicated that the two inhibitory peptides (as well as several non-inhibitory peptides) interact directly with IN. Moreover, the isolated catalytic core domain of IN also interacted directly with the two inhibitory peptides. Nevertheless, only peptide 4286 can inhibit the disintegration activity associated with the IN core domain, because this activity is the only one exhibited by this domain. This result was expected from the lack of inhibition of disintegration of full-length IN by peptide 4321. The data and the three-dimensional models presented suggested that the inhibition resulted from steric hindrance of the catalytic domain of IN. This information can substantially facilitate the development of novel drugs against HIV INs and thus contribute to the fight against AIDS. PMID- 15790560 TI - Transcriptional regulation of NF-E2 p45-related factor (NRF2) expression by the aryl hydrocarbon receptor-xenobiotic response element signaling pathway: direct cross-talk between phase I and II drug-metabolizing enzymes. AB - The aryl hydrocarbon receptor (AHR) and NF-E2 p45-related factor (NRF2) are two distinct transcription factors involved in the regulation of drug-metabolizing enzymes. Increasing evidence from several studies implies that AHR and NRF2 have direct links, but the molecular mechanism remains unknown. In this work we demonstrate for the first time that Nrf2 gene transcription is directly modulated by AHR activation. DNA sequence analyses of the mouse Nrf2 promoter revealed one xenobiotic response element (XRE)-like element (XREL1) located at -712 and two additional XRE-like elements located at +755 (XREL2) and +850 (XREL3). Functional analysis using luciferase assay showed that XREL1, XREL2, and XREL3 are all inducible by 2,3,7,8-tetrachlorodibenzo-p-dioxin treatment, with XREL2 being the most potent. The functionality of these XRE-like elements was further confirmed by mutagenesis and gel shift experiments. Finally, we used chromatin immunoprecipitation assay to show a direct binding of AHR to the Nrf2 promoter. Cells with silenced AHR expression using siRNA also lost NRF2 mRNA induction by 2,3,7,8-tetrachlorodibenzo-p-dioxin. These new data position NRF2-antioxidant response element downstream in the AHR-XRE pathway. Moreover, direct regulation of NRF2 by AHR contributes to couple phase I and II enzymes into an integrated system facilitating more effective xenobiotic and carcinogen detoxification. PMID- 15790561 TI - Regulation of N-methyl-D-aspartate receptors by calpain in cortical neurons. AB - The N-methyl-D-aspartate (NMDA) receptor is a cation channel highly permeable to calcium and plays critical roles in governing normal and pathologic functions in neurons. Calcium entry through NMDA receptors (NMDARs) can lead to the activation of the Ca2+-dependent protease, calpain. Here we investigated the involvement of calpain in regulation of NMDAR channel function. After prolonged (5-min) treatment with NMDA or glutamate, the whole-cell NMDAR-mediated current was significantly reduced in both acutely dissociated and cultured cortical pyramidal neurons. The down-regulation of NMDAR current was blocked by bath application of selective calpain inhibitors. Intracellular injection of a specific calpain inhibitory peptide also eliminated the down-regulation of NMDAR current induced by prolonged NMDA treatment. In contrast, dynamin inhibitory peptide had no effect on the depression of NMDAR current, suggesting the lack of involvement of dynamin/clathrin-mediated NMDAR internalization in this process. Immunoblotting analysis showed that the NR2A and NR2B subunits of NMDARs were markedly degraded in cultured cortical neurons treated with glutamate, and the degradation of NR2 subunits was prevented by calpain inhibitors. Taken together, our results suggest that prolonged activation of NMDARs in neurons activates calpain, and activated calpain in turn down-regulates the function of NMDARs, which provides a neuroprotective mechanism against NMDAR overstimulation accompanying ischemia and stroke. PMID- 15790564 TI - Biochemical characterization of the Campylobacter jejuni Cj1294, a novel UDP-4 keto-6-deoxy-GlcNAc aminotransferase that generates UDP-4-amino-4,6-dideoxy GalNAc. AB - Campylobacter jejuni produces multiple glycoproteins whose glycans contain 4 amino 6-deoxy sugars or their derivatives, such as diacetamidobacillosamine or pseudaminic acid. Because the proteoglycans contribute to bacterial virulence and their constitutive sugars are not commonly found in humans, inhibitors developed against the enzymes that are responsible for their biosynthesis could be novel therapeutic targets to fight this important food-borne pathogen. The biosynthesis of diacetamidobacillosamine is anticipated to involve a sugar nucleotide C6 dehydratase, a C4 aminotransferase and an acetyltransferase. We have identified a set of genes (cj1293, cj1294, and cj1298) potentially encoding one of each enzymatic activity, and demonstrated earlier that Cj1293 was a UDP-GlcNAc specific C6 dehydratase. Others have shown that Cj1293 was involved in protein glycosylation. Here, we report on our investigation of the potential activity of Cj1294 as a sugar nucleotide C4 aminotransferase. Our biochemical characterization of overexpressed and purified protein shows that Cj1294 is a pyridoxal phosphate-dependent aminotransferase specific for UDP-4-keto-6-deoxy GlcNAc that uses preferentially glutamic acid as an amino donor. A detailed physicokinetic study of Cj1294 was performed to determine the K(m) of 1.28 +/- 0.2 mm and k(cat) of 11.5 +/- 1.3 min(-1). Also, two residues essential for protein stability and activity, Arg(228) and Lys(181), respectively, were identified by site-directed mutagenesis. Finally, we demonstrated by NMR analysis of purified reaction product that Cj1294 produces UDP-4-amino-4,6-dideoxy-GalNAc. These results indicate that Cj1294 is involved in the biosynthesis of diacetamidofucosamine, a C4 epimer of diacetamidobacillosamine not yet described in C. jejuni proteoglycans, suggesting that the composition of C. jejuni proteoglycans is more variable than anticipated. PMID- 15790563 TI - Neurochondrin negatively regulates CaMKII phosphorylation, and nervous system specific gene disruption results in epileptic seizure. AB - Neurochondrin is a novel cytoplasmic protein and possibly involved in neurite outgrowth, chondrocyte differentiation, and bone metabolism. Our previous trial in disclosing its role by the loss of function in mice failed because of the lethality in utero. In this study, we eliminated the neurochondrin gene expression preferentially in the nervous system by the conditional knockout strategy. Our results showed that neurochondrin is a negative regulator of Ca(2+)/calmodulin-dependent protein kinase II phosphorylation and essential for the spatial learning process but not for the differentiation or neurite outgrowth of the neuron. In addition, the nervous system-specific homozygous gene disruption resulted in epileptic seizure. PMID- 15790562 TI - Lys6-modified ubiquitin inhibits ubiquitin-dependent protein degradation. AB - Ubiquitin plays essential roles in various cellular processes; therefore, it is of keen interest to study the structure-function relationship of ubiquitin itself. We investigated the modification of Lys(6) of ubiquitin and its physiological consequences. Mass spectrometry-based peptide mapping and N terminal sequencing demonstrated that, of the 7 Lys residues in ubiquitin, Lys(6) was the most readily labeled with sulfosuccinimidobiotin. Lys(6)-biotinylated ubiquitin was incorporated into high molecular mass ubiquitin conjugates as efficiently as unmodified ubiquitin. However, Lys(6)-biotinylated ubiquitin inhibited ubiquitin-dependent proteolysis, as conjugates formed with Lys(6) biotinylated ubiquitin were resistant to proteasomal degradation. Ubiquitins with a mutation of Lys(6) had similar phenotypes as Lys(6)-biotinylated ubiquitin. Lys(6) mutant ubiquitins (K6A, K6R, and K6W) also inhibited ATP-dependent proteolysis and caused accumulation of ubiquitin conjugates. Conjugates formed with K6W mutant ubiquitin were also resistant to proteasomal degradation. The dominant-negative effect of Lys(6)-modified ubiquitin was further demonstrated in intact cells. Overexpression of K6W mutant ubiquitin resulted in accumulation of intracellular ubiquitin conjugates, stabilization of typical substrates for ubiquitin-dependent proteolysis, and enhanced susceptibility to oxidative stress. Taken together, these results show that Lys(6)-modified ubiquitin is a potent and specific inhibitor of ubiquitin-mediated protein degradation. PMID- 15790565 TI - Functional characterization of mouse RDH11 as a retinol dehydrogenase involved in dark adaptation in vivo. AB - We previously cloned mouse RDH11 (mRDH11) as a gene regulated by the transcription factor sterol regulatory element-binding proteins and showed that it is a retinol dehydrogenase expressed in non-ocular tissues such as the liver and testis and in the retina (Kasus-Jacobi, A., Ou, J., Bashmakov, Y. K., Shelton, J. M., Richardson, J. A., Goldstein, J. L., and Brown, M. S. (2003) J. Biol. Chem. 278, 32380-32389). It was proposed to function in the recycling of the visual chromophore 11-cis-retinal after photoisomerization by a bleaching light, a pathway referred to as the visual cycle. In this work, we describe our studies on the ocular function of mRDH11. We created a knockout mouse by replacing the mrdh11 coding sequence with the lacZ reporter gene for expression profiling. 5-Bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining demonstrated active transcription of this gene in photoreceptor cells. We show by immunoblot analysis that mRDH11 is associated with retinal membranes purified from a non-outer segment fraction of the retina. No obvious retinal defect was found during development and aging of RDH11-deficient mice. The functional consequences of mRDH11 disruption were investigated by electroretinography. Dark adaptation was delayed by a factor of 2.5-3 compared with wild-type mice. However, the kinetics of 11-cis-retinal recycling during dark adaptation was not affected, suggesting that mRDH11 is not involved in the visual cycle. We propose that mRDH11 disruption affects retinoid metabolism in photoreceptor inner segments and delays the kinetics of dark adaptation through modulation of calcium homeostasis. PMID- 15790566 TI - Cdk5 activator-binding protein C53 regulates apoptosis induced by genotoxic stress via modulating the G2/M DNA damage checkpoint. AB - In response to DNA damage, the cellular decision of life versus death involves an intricate network of multiple factors that play critical roles in regulation of DNA repair, cell cycle, and cell death. DNA damage checkpoint proteins are crucial for maintaining DNA integrity and normal cellular functions, but they may also reduce the effectiveness of cancer treatment. Here we report the involvement of Cdk5 activator p35-binding protein C53 in regulation of apoptosis induced by genotoxic stress through modulating Cdk1-cyclin B1 function. C53 was originally identified as a Cdk5 activator p35-binding protein and a caspase substrate. Importantly, our results demonstrated that C53 deficiency conferred partial resistance to genotoxic agents such as etoposide and x-ray irradiation, whereas ectopic expression of C53 rendered cells susceptible to multiple genotoxins that usually trigger G(2)/M arrest. Furthermore, we found that Cdk1 activity was required for etoposide-induced apoptosis of HeLa cells. Overexpression of C53 promoted Cdk1 activity and nuclear accumulation of cyclin B1, whereas C53 deficiency led to more cytoplasmic retention of cyclin B1, suggesting that C53 acts as a pivotal player in modulating the G(2)/M DNA damage checkpoint. Finally, C53 and cyclin B1 co-localize and associate in vivo, indicating a direct role of C53 in regulating the Cdk1-cyclin B1 complex. Taken together, our results strongly indicate that in response to genotoxic stress, C53 serves as an important regulatory component of the G(2)/M DNA damage checkpoint. By overriding the G(2)/M checkpoint-mediated inhibition of Cdk1-cyclin B1 function, ectopic expression of C53 may represent a novel approach for chemo- and radio sensitization of cancer cells. PMID- 15790567 TI - Chemical synthesis of (S)-4,5-dihydroxy-2,3-pentanedione, a bacterial signal molecule precursor, and validation of its activity in Salmonella typhimurium. AB - We describe an original, short, and convenient chemical synthesis of enantiopure (S)-4,5-dihydroxy-2,3-pentanedione (DPD), starting from commercial methyl (S)-(-) 2,2-dimethyl-1,3-dioxolane-4-carboxylate. DPD is the precursor of autoinducer (AI)-2, the proposed signal for bacterial interspecies communication. AI-2 is synthesized by many bacterial species in three enzymatic steps. The last step, a LuxS-catalyzed reaction, leads to the formation of DPD, which spontaneously cyclizes into AI-2. AI-2-like activity of the synthesized molecule was ascertained by the Vibrio harveyi bioassay. To further validate the biological activity of synthetic DPD and to explore its potential in studying DPD (AI-2) mediated signaling, a Salmonella typhimurium luxS mutant was constructed. Expression of the AI-2 regulated lsr operon can be rescued in this luxS mutant by addition of synthetic DPD or genetic complementation. Biofilm formation by S. typhimurium has been reported to be defective in a luxS mutant, and this was confirmed in this study to test DPD for chemical complementation. However, biofilm formation of the luxS mutant cannot be restored by addition of DPD. In contrast, introduction of luxS under control of its own promoter complemented biofilm formation. Further results demonstrated that biofilm formation of the luxS mutant cannot be restored with luxS under control of the strong nptII promoter. This indicates that altering the intrinsic promoter activity of luxS affects Salmonella biofilm formation. Conclusively, we synthesized biologically active DPD. Using this chemical compound in combination with genetic approaches opens new avenues in studying AI-2-mediated signaling. PMID- 15790568 TI - The role of EF-hand domains and C2 domain in regulation of enzymatic activity of phospholipase Czeta. AB - Sperm-specific phospholipase C-zeta (PLCzeta) induces Ca2+ oscillations and egg activation when injected into mouse eggs. PLCzeta has such a high Ca2+ sensitivity of PLC activity that the enzyme can be active in resting cells at approximately 100 nM Ca2+, suitable for a putative sperm factor to be introduced into the egg at fertilization (Kouchi, Z., Fukami, K., Shikano, T., Oda, S., Nakamura, Y., Takenawa, T., and Miyazaki, S. (2004) J. Biol. Chem. 279, 10408 10412). In the present structure-function analysis, deletion of EF1 and EF2 of the N-terminal four EF-hand domains caused marked reduction of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2)-hydrolyzing activity in vitro and loss of Ca2+ oscillation-inducing activity in mouse eggs after injection of RNA encoding the mutant. However, deletion of EF1 and EF2 or mutation of EF1 or EF2 at the x and z positions of the putative Ca2+-binding loop little affected the Ca2+ sensitivity of the PLC activity, whereas deletion of EF1 to EF3 caused 12-fold elevation of the EC50 of Ca2+ concentration. Thus, EF1 and EF2 are important for the PLCzeta activity, and EF3 is responsible for its high Ca2+ sensitivity. Deletion of four EF-hand domains or the C-terminal C2 domain caused complete loss of PLC activity, indicating that both regions are prerequisites for PLCzeta activity. Screening of interactions between the C2 domain and phosphoinositides revealed that C2 has substantial affinity to PI(3)P and, to the lesser extent, to PI(5)P but not to PI(4,5)P2 or acidic phospholipids. PI(3)P and PI(5)P reduced PLCzeta activity in vitro, suggesting that the interaction could play a role for negative regulation of PLCzeta. PMID- 15790569 TI - Identification of estrogen-responsive genes using a genome-wide analysis of promoter elements for transcription factor binding sites. AB - We developed a pipeline to identify novel genes regulated by the steroid hormone dependent transcription factor, estrogen receptor, through a systematic analysis of upstream regions of all human and mouse genes. We built a data base of putative promoter regions for 23,077 human and 19,984 mouse transcripts from National Center for Biotechnology Information annotation and 8793 human and 6785 mouse promoters from the Data Base of Transcriptional Start Sites. We used this data base of putative promoters to identify potential targets of estrogen receptor by identifying estrogen response elements (EREs) in their promoters. Our program correctly identified EREs in genes known to be regulated by estrogen in addition to several new genes whose putative promoters contained EREs. We validated six genes (KIAA1243, NRIP1, MADH9, NME3, TPD52L, and ABCG2) to be estrogen-responsive in MCF7 cells using reverse transcription PCR. To allow for extensibility of our program in identifying targets of other transcription factors, we have built a Web interface to access our data base and programs. Our Web-based program for Promoter Analysis of Genome, PAGen@UIC, allows a user to identify putative target genes for vertebrate transcription factors through the analysis of their upstream sequences. The interface allows the user to search the human and mouse promoter data bases for potential target genes containing one or more listed transcription factor binding sites (TFBSs) in their upstream elements, using either regular expression-based consensus or position weight matrices. The data base can also be searched for promoters harboring user-defined TFBSs given as a consensus or a position weight matrix. Furthermore, the user can retrieve putative promoter sequences for any given gene together with identified TFBSs located on its promoter. Orthologous promoters are also analyzed to determine conserved elements. PMID- 15790571 TI - Solution structure of the C-terminal domain of TFIIH P44 subunit reveals a novel type of C4C4 ring domain involved in protein-protein interactions. AB - The human general transcription factor TFIIH is involved in both transcription and DNA nucleotide excision repair. Among the 10 subunits of the complex, p44 subunit plays a crucial role in both mechanisms. Its N-terminal domain interacts with the XPD helicase, whereas its C-terminal domain is involved specifically in the promoter escape activity. By mutating an exposed and non-conserved cysteine residue into a serine, we produced a soluble mutant of p44-(321-395) suitable for solution structure determination. The domain adopts a C4C4 RING domain structure with sequential organization of beta-strands that is related to canonical RING domains by a circular permutation of the beta-sheet elements. Analysis of the molecular surface and mutagenesis experiments suggests that the binding of p44 (321-395) to TFIIH p34 subunit is not mediated by electrostatic interactions and, thus, differs from previously reported interaction mechanisms involving RING domains. PMID- 15790570 TI - Activation of p38 has opposing effects on the proliferation and migration of endothelial cells. AB - Pathological conditions such as hypertension and hyperglycemia as well as abrasions following balloon angioplasty all lead to endothelial dysfunction that impacts disease morbidity. These conditions are associated with the elaboration of a variety of cytokines and increases in p38 activity in endothelial cells. However, the relationship between enhanced p38 activity and endothelial cell function remains poorly understood. To investigate the effect of enhanced p38 MAPK activity on endothelial cell function, we expressed an activated mutant of MEK6 (MEK6E), an upstream regulator of p38. Expression of MEK6E activated p38 and resulted in phosphorylation of its downstream substrate, heat shock protein 27 (Hsp27). Activation of p38 was not sufficient to induce apoptosis; however, it did induce p38-dependent cell cycle arrest. MEK6E expression was sufficient to inhibit ERK phosphorylation triggered by growth factors and integrin engagement. MAPK phosphatase-1 (MKP-1) expression was increased upon p38 activation, and expression of a "substrate-trapping" MKP-1 was sufficient to restore ERK activity. Activation of p38 was sufficient to induce cell migration, which was accompanied by alterations in actin architecture characterized by enhanced lamellipodia. Co-expression of a mutant form of Hsp27, lacking all three phosphorylation sites, reversed MEK6E-induced cell migration and altered the cytoskeletal changes induced by p38 activation. Collectively, these results suggest that cellular decisions regarding migration and proliferation are influenced by p38 activity and that prolonged activation of p38 may result in an anti-angiogenic phenotype that contributes to endothelial dysfunction. PMID- 15790572 TI - Engineering and functional immobilization of opioid receptors. AB - Opioid receptors, like many G protein-coupled receptors (GPCRs), are notoriously unstable in detergents. We have now developed a more stable variant of the mu opioid receptor (MOR) and also a method for the immobilization of solubilized, functional opioid receptors on a solid phase (magnetic beads). Starting with the intrinsically more stable kappa-opioid receptor (KOR), we optimized the conditions (i.e. detergents and stabilizing ligands) for receptor extraction from lipid bilayers of HEK293T cells to obtain maximal amounts of functional, immobilized receptor. After immobilization, the ligand binding profile remains the same as observed for the membrane-embedded receptor. For the immobilized wild type mu-opioid receptor, however, no conditions were found under which ligand binding capacity was retained. To solve this problem, we engineered the receptor chimera KKM where the N-terminus and the first transmembrane helix (TM1) of wild type MOR is exchanged for the homologous receptor parts of the wild-type KOR. This hybrid receptor behaves exactly as the wild-type MOR in functional assays. Interestingly, the modified MOR is expressed at six times higher levels than wild type MOR and is similarly stable as wild-type KOR after immobilization. Hence the immobilized MOR, represented by the chimera KKM, is now also amenable for biophysical characterization. These results are encouraging for future stability engineering of GPCRs. PMID- 15790573 TI - Peptide inhibitors of the essential cell division protein FtsA. AB - The revolutionary era of antibiotics has been overwhelmed by the evolutionary capacity of microorganisms such as Pseudomonas aeruginosa to develop resistance to all classes of antibiotics. In the perspective of identifying new antimicrobials using novel strategies, we targeted the essential and highly conserved FtsA protein from the bacterial cell division machinery of P.aeruginosa. In a series of experiments we cloned, overproduced and purified the FtsA and FtsZ proteins. Expression of FtsA into Escherichia coli cells led to its accumulation in inclusion bodies. We developed a protocol permitting the purification and refolding of enzymatically active FtsA hydrolysing ATP. The purified enzyme was used to screen for peptide inhibitors of ATPase activity using phage display. Selective biopanning assays were done and phages were eluted using ATP, a non-hydrolysable ATP analogue and the protein FtsZ known to interact with FtsA in the divisome during the process of bacterial cell division. We identified two consensus peptide sequences interacting with FtsA and a competitive ELISA was used to identify peptides having high affinity for the target protein. Five of the six peptides synthesized showed specific inhibition of ATPase activity of FtsA with IC50 values between 0.7 and 35 mM. Discovery of peptides inhibiting the essential cell division machinery in bacteria is the first step for the future development of antimicrobial agents via peptidomimetism. PMID- 15790574 TI - G protein-coupled receptors show unusual patterns of intrinsic unfolding. AB - Intrinsically unstructured proteins (IUPs) or IUP-like regions often play key roles in controlling processes ranging from transcription to the cell cycle. In silico such proteins can be identified by their sequence properties; they have low hydrophobicity and high net charge. In this study, we applied the FoldIndex (http://bioportal.weizmann.ac.il/fldbin/findex) program to analyze human G protein-coupled receptors and compared them with membrane proteins of known structure and with IUPs. We show that human G protein-coupled receptor (GPCR) extramembranous domains include long (>50 residues) disordered segments, unlike membrane proteins of known structure. The predicted disorder occurred primarily in the N-terminal, C-terminal and third intracellular domain regions: 55, 69 and 56% of the human GPCRs were disordered in these regions, respectively. This increased flexibility may therefore be critical for GPCR function. Surprisingly, however, the kinds of residues used in GPCR unstructured regions were different than in hitherto-identified IUPs. The GPCR third intracellular loop domains contain very high percentages of Arg, Lys and His residues, especially Arg, but the percentage of Glu, Asp and Pro is no higher than in folded proteins. We propose that this has structural and functional consequences. PMID- 15790575 TI - Engineering a pharmacologically superior form of leptin for the treatment of obesity. AB - Leptin plays a central role in the homeostasis of body weight through its regulatory effects on appetite and energy expenditure, yet in trials as a therapeutic agent for the treatment of obesity in humans it has been disappointing. The poor clinical efficacy of leptin results from its short circulating half-life, low potency and poor solubility, necessitating large and frequent doses to obtain even modest clinical benefit. Engineered Fc-leptin immunofusins, consisting of the Fc fragment of an immunoglobulin gamma chain followed by leptin, exhibit improved pharmacological properties with very consistent and potent biological activities. Furthermore, in extending the circulating half-life of the protein in vivo from a few minutes for leptin to many hours for Fc-leptin, these proteins have the potential to reduce drastically the dosage and frequency of administration required to obtain clinical benefit. The results of this study show that the engineered leptin immunofusins described here have significantly enhanced pharmacological properties in comparison with the recombinant leptin that was used in clinical trials. As such, they could represent an important step towards a therapeutically superior form of leptin if the disappointing performance of leptin in early clinical trials was due to its poor pharmacological properties rather than any conceptual weakness in the strategy of using leptin for the treatment of obesity and its related disorders. PMID- 15790576 TI - Favorable scaffolds: proteins with different sequence, structure and function may associate in similar ways. AB - Proteins with similar structures may have different functions. Here, using a non redundant two-chain protein-protein interface dataset containing 103 clusters, we show that this paradigm extends to interfaces. Whereas usually similar interfaces are obtained from globally similar chains, this is not always the case. Remarkably, in some interface clusters, although the interfaces are similar, the overall structures and functions of the chains are different. Hence, our work suggests that different folds may combinatorially assemble to yield similar local interface motifs. The preference of different folds to associate in similar ways illustrates that the paradigm is universal, whether for single chains in folding or for protein-protein association in binding. We analyze and compare the two types of clusters. Type I, with similar interfaces, similar global structures and similar functions, is better packed, less planar, has larger total and non-polar buried surface areas, better complementarity and more backbone-backbone hydrogen bonds than Type II (similar interfaces, different global structures and different functions). The dataset clusters may provide rich data for protein-protein recognition, cellular networks and drug design. In particular, they should be useful in addressing the difficult question of what the favorable ways for proteins to interact are. PMID- 15790577 TI - Bioactive IL7-diphtheria fusion toxin secreted by mammalian cells. AB - A number of targeted cytotoxic agents have been developed that selectively kill malignant or otherwise pathological cells. These engineered proteins consist of a potent cytotoxic element connected to a ligand domain that binds to specific molecules on the surface of the target cell. Several of these agents have shown promise in clinical trials and one is currently administered to patients. A significant technical obstacle that has impeded the development of some of these toxins is the difficulty of preparing certain recombinant proteins in properly folded forms. These fusion proteins have generally been produced in bacteria requiring them to be denatured and renatured in vitro. For some proteins this is an efficient process whereas for others it is not. We describe here a system to produce fusion toxins rapidly and efficiently by engineering mammalian cells to secrete them as properly folded molecules which can be purified in native form from cell culture medium. We have used this system to produce highly active preparations of DAB(389)-IL7, a molecule consisting of the catalytic and transmembrane domains of diphtheria toxin fused to interleukin 7. This system is generalizable and can be used to produce and evaluate rapidly fusion toxins incorporating novel or uncharacterized ligands. PMID- 15790578 TI - Engineering of a thioglycoligase: randomized mutagenesis of the acid-base residue leads to the identification of improved catalysts. AB - Thioglycoligases are recently introduced variants of retaining glycosidases in which the acid-base catalyst has been mutated, rendering them capable of thioglycoside synthesis. The original acid-base mutant of Agrobacterium sp. beta glucosidase (E170A) was previously shown to be an effective thioglycoligase carrying out glycosyltransfer from 2,4-dinitrophenyl glycosides to several different thio sugar acceptors. Here we report the generation of a screen for improved thioglycoligases, randomized mutagenesis of the acid-base catalyst E170 and identification of variants superior to E170A. Furthermore we have established a coupled assay allowing kinetic analysis of isolated variants and found that Abg E170Q is 5-fold faster than Abg E170A when 2,4-dinitrophenyl glucoside is used as donor and 100-fold faster when glucosyl azide is used. To demonstrate its utility, different acceptor and donor sugar combinations were employed to produce thio-linked di- or trisaccharides in high yields, showing the considerable versatility of the system for the synthesis of carbohydrate mimetics. PMID- 15790579 TI - Helix stability and hydrophobicity in the folding mechanism of the bacterial immunity protein Im9. AB - Recent models suggest that the mechanism of protein folding is determined by the balance between the stability of secondary structural elements and the hydrophobicity of the sequence. Here we determine the role of these factors in the folding kinetics of Im9* by altering the secondary structure propensity or hydrophobicity of helices I, II or IV by the substitution of residues at solvent exposed sites. The folding kinetics of each variant were measured at pH 7.0 and 10 degrees C, under which conditions wild-type Im9* folds with two-state kinetics. We show that increasing the helicity of these sequences in regions known to be structured in the folding intermediate of Im7*, switches the folding of Im9* from a two- to three-state mechanism. By contrast, increasing the hydrophobicity of helices I or IV has no effect on the kinetic folding mechanism. Interestingly, however, increasing the hydrophobicity of solvent-exposed residues in helix II stabilizes the folding intermediate and the rate-limiting transition state, consistent with the view that this helix makes significant non-native interactions during folding. The results highlight the generic importance of intermediates in folding and show that such species can be populated by increasing helical propensity or by stabilizing inter-helix contacts through non native interactions. PMID- 15790580 TI - Directed evolution of phosphotriesterase from Pseudomonas diminuta for heterologous expression in Escherichia coli results in stabilization of the metal free state. AB - Phosphotriesterase from Pseudomonas diminuta (PTE) is an extremely efficient metalloenzyme that hydrolyses a variety of compounds including organophosphorus nerve agents. Study of PTE has been hampered by difficulties with efficient expression of the recombinant form of this highly interesting and potentially useful enzyme. We identified a low-level esterolytic activity of PTE and then screened PTE gene libraries for improvements in 2-naphthyl acetate hydrolysis. However, the attempt to evolve this promiscuous esterase activity led to a variant (S5) containing three point mutations that resulted in a 20-fold increase in functional expression. Interestingly, the zinc holoenzyme form of S5 appears to be more sensitive than wild-type PTE to both thermal denaturation and addition of metal chelators. Higher functional expression of the S5 variant seems to lie in a higher stability of the metal-free apoenzyme. The results obtained in this work point out another-and often overlooked-possible determinant of protein expression and purification yields, i.e. the stability of intermediates during protein folding and processing. PMID- 15790581 TI - How to predict response to cardiac resynchronization therapy? PMID- 15790582 TI - Advances in understanding the mechanisms of angina pectoris in cardiac syndrome X. PMID- 15790583 TI - Abnormal cortical pain processing in patients with cardiac syndrome X. AB - AIMS: Previous studies suggested that an enhanced pain sensitivity is present in patients with cardiac syndrome X (SX). We investigated whether SX patients present abnormalities in the electrical cerebral signals generated by pain stimuli. METHODS AND RESULTS: Cortical laser evoked potentials (LEPs) were recorded in 16 SX patients, in 10 patients with refractory angina due to obstructive coronary artery disease (CAD) and in 13 healthy controls. LEPs were recorded during stimulation of chest and right hand dorsum. Three sequences of painful stimuli were applied at each site. Subjective pain rating was assessed by a 0-100 mm visual analogic scale (VAS). Basal LEPs did not differ among groups and there were no differences for most LEP components across the repetitions of stimuli. However, the amplitude of the N2/P2 LEP component, specifically reflecting cortical pain processing, decreased across the three sequences of stimuli in controls and CAD patients, but not in SX patients. Compared with the first sequence, the N2/P2 amplitude during the third sequence of stimuli in the three groups was 77+/-16, 56+/-24, and 99+/-34%, respectively, for chest (P=0.001), and 63+/-31, 72+/-17, and 98+/-46%, respectively, for right hand (P=0.03) stimulation. The changes in VAS pain score across the three sequences paralleled those of N2/P2 amplitude. CONCLUSION: Our data show that in SX patients, central handling of painful stimuli is characterized by inadequate habituation, which might play a role in determining the peculiar clinical characteristics of anginal chest pain of these patients. PMID- 15790584 TI - Applicability and limitations of optimal biomass allocation models: a test of two species from fertile and infertile habitats. AB - BACKGROUND AND AIMS: The practical applicability of optimal biomass allocation models is not clear. Plants may have constraints in the plasticity of their root : leaf ratio that prevent them from regulating their root : leaf ratio in the optimal manner predicted by the models. The aim of this study was to examine the applicability and limitations of optimal biomass allocation models and to test the assumption that regulation of the root : leaf ratio enables maximization of the relative growth rate (RGR). METHODS: Polygonum cuspidatum from an infertile habitat and Chenopodium album from a fertile habitat were grown under a range of nitrogen availabilities. The biomass allocation, leaf nitrogen concentration (LNC), RGR, net assimilation rate (NAR), and leaf area ratio (LAR) of each species were compared with optimal values determined using an optimal biomass allocation model. KEY RESULTS: The root : leaf ratio of C. album was smaller than the optimal ratio in the low-nitrogen treatment, while it was almost optimal in the high-nitrogen treatment. In contrast, the root : leaf ratio of P. cuspidatum was close to the optimum under both high- and low-nitrogen conditions. Owing to the optimal regulation of the root : leaf ratio, C. album in the high-nitrogen treatment and P. cuspidatum in both treatments had LNC and RGR (with its two components, NAR and LAR) close to their optima. However, in the low-nitrogen treatment, the suboptimal root : leaf ratio of C. album led to a smaller LNC than the optimum, which in turn resulted in a smaller NAR than the optimum and RGR than the theoretical maximum RGR. CONCLUSIONS: The applicability of optimal biomass allocation models is fairly high, although constraints in the plasticity of biomass allocation could prevent optimal regulation of the root : leaf ratio in some species. The assumption that regulation of the root : leaf ratio enables maximization of RGR was supported. PMID- 15790585 TI - Genetic variation in remnant populations of Dalbergia nigra (Papilionoideae), an endangered tree from the Brazilian Atlantic Forest. AB - BACKGROUND AND AIMS: Dalbergia nigra, known as Brazilian rosewood, is an endangered tree species restricted to the Brazilian Atlantic Forest and has been intensively logged for five centuries due to its high-quality wood. The objective of the present study was to assess the genetic variation and structure in adults and saplings of the species from a large reserve of the Atlantic Forest, the Rio Doce State Park, and from two small surrounding fragments, one better preserved and another with a high degree of anthropogenic disturbance. METHODS: Analyses of genetic variation and structure were conducted by studying allozyme markers. Seven putative enzymatic loci were resolved, five of them being polymorphic. KEY RESULTS: The mean numbers of alleles per locus (A) were 1.93 and 1.73, while the percentages of polymorphic loci (P) were 93 and 73 % for adults and saplings, respectively. Saplings from the fragment with high anthropogenic disturbance exhibited the lowest values of A and P. The fragment that constitutes a conservation area exhibited genetic variation similar to the population from the large reserve. The observed (H(o)) and expected (H(e)) heterozygosities were not significantly different among the three populations. Only sapling populations showed F(ST) values (divergence among populations) significantly different from zero over all studied loci. The fragment with high anthropogenic disturbance exhibited considerable genetic divergence in relation to the above-cited populations. CONCLUSIONS: The evaluated populations displayed mean levels of genetic variation intermediate to those expected for narrow and widespread species. The results suggest that fragments with similar area and geographical distance from a large protected reserve can exhibit different levels of genetic variation, depending on the degree of anthropogenic disturbance. The considerable genetic variation in the protected fragment points to the importance of adequate conservation of small fragments for the preservation of genetic variation in D. nigra. PMID- 15790586 TI - Molecular variation and fingerprinting of Leucadendron cultivars (Proteaceae) by ISSR markers. AB - BACKGROUND AND AIMS: There are more than 80 species of Leucadendron and most are used as cut flowers. Currently, more than 100 cultivars are used by industry and many of them are interspecific hybrids. The origin of most cultivars is unclear and their genetic diversity and relationships have not been studied. This investigation was carried out to evaluate the genetic variation and relationships among 30 Leucadendron cultivars. METHODS: ISSR markers were applied to determine the genetic variation and to discriminate Leucadendron cultivars. Sixty-four ISSR primers were screened and 25 primers were selected for their ability to produce clear and reproducible patterns of multiple bands. KEY RESULTS: A total of 584 bands of 305-2400 bp were amplified, of which 97 % were polymorphic. A dendrogram generated using the Unweighted Pair Group Method with Arithmetic Average based on a distance measure of total character difference showed that the Leucadendron cultivars clustered into two main groups. Twenty-four of the 30 cultivars can be unequivocally differentiated, but identical profiles were observed for three cultivar pairs, 'Katie's Blush' and 'Silvan Red', 'Highlights' and 'Maui Sunset', and 'Yellow Crest' and 'Yellow Devil'. CONCLUSIONS: ISSR profiling is a powerful method for the identification and molecular classification of Leucadendron cultivars. A fingerprinting key was generated based on the banding patterns produced using two ISSR primers (UBC856 and UBC857). In addition cultivar specific ISSR bands were obtained for 17 of the 30 Leucadendron cultivars tested. PMID- 15790587 TI - Gene-environment interactions between alcohol drinking and the MTHFR C677T polymorphism impact on esophageal cancer risk: results of a case-control study in Japan. AB - Folate takes part in two biological pathways involved in DNA methylation and synthesis, and a potential protective influence of this nutrient chemical against carcinogenicity has been recognized in several sites, including the esophagus. Therefore, the functional polymorphisms in genes encoding folate metabolizing enzymes, MTHFR C677T and MTR A2756G, might be suspected of impacting on esophageal cancer risk. We therefore conducted a matched case-control study of 165 esophageal cancer cases and 495 non-cancer controls to clarify associations among folate intake, MTHFR C677T and MTR A2756G polymorphisms, and esophageal cancer risk. Gene-environment interactions between the two polymorphisms, and drinking and smoking were also evaluated. Folate consumption and MTHFR 677TT were associated with a non-significant tendency for decreased risk while the MTR genotypes did not show any links in themselves; further, when analysis was limited to heavy drinkers, the MTHFR TT genotype significantly decreased esophageal cancer risk [odds ratio (OR) = 0.27, 95% confidence interval (CI), 0.09-0.76]. The OR for the gene-environment interaction between heavy drinking and the 677TT genotype in the case-only design was 0.31 (95% CI, 0.10-0.94), indicating risk with heavy drinking to be 69% decreased in individuals harboring the 677TT genotype. We failed to find any significant interaction between either of the polymorphisms and smoking. PMID- 15790588 TI - Deregulated expression of the PER1, PER2 and PER3 genes in breast cancers. AB - Disruption of circadian rhythm may be a risk factor in the development of breast cancer, but molecular changes in circadian rhythm controlled genes in breast cancer cells are still unexplored. We used immunohistochemical staining, methylation specific PCR and direct sequencing methods to analyze molecular changes in three most important genes, namely PER1, PER2 and PER3, in circadian rhythm in 55 cases of breast cancer of Taiwanese women. Our results reveal disturbances in the expression of the three period (PER) genes in most (>95%) of the breast cancerous cells in comparison with the nearby non-cancerous cells. The PER gene deregulation is not caused by genetic mutations but most probably by methylation of the PER1 or PER2 promoter. Methylation of the PER gene promoters has a strong correlation with c-erbB2 expression (P = 0.017). Since the circadian clock controls expression of cell-cycle related genes, we suggest that disturbances in PER gene expression may result in disruption of the control of the normal circadian clock, thus benefiting the survival of cancer cells and promoting carcinogenesis. Differential expression of circadian genes in non cancerous and cancerous cells may provide a molecular basis for chronotherapy of breast cancer. PMID- 15790589 TI - Chemopreventive properties of apple procyanidins on human colon cancer-derived metastatic SW620 cells and in a rat model of colon carcinogenesis. AB - Apples contain several classes of polyphenols: monomers (catechins, epicatechins) and oligomers/polymers, such as the procyanidins. Our aim was (i) to study anti proliferative mechanisms on human metastatic colon carcinoma (SW620 cells) of apple polyphenol fractions (monomers or procyanidins) and (ii) to evaluate their anti-carcinogenic properties in vivo. Two polyphenol-enriched fractions were isolated from apples. Fraction non-procyanidins contained 73% phenolic monomers and no procyanidins, while fraction procyanidins contained 78% procyanidins and no monomers. Inhibition of SW620 cell growth was only observed with fraction P (IC50 = 45 microg/ml). After a 24-h exposure of cells to fraction P, protein kinase C activity was inhibited by 70% and a significant increase in extracellular signal-regulated kinases 1 and 2 and c-jun N-terminal kinases expression was observed together with the down-regulation of polyamine biosynthesis and the activation of caspase-3. Colon carcinogenesis was induced in rats by intraperitoneal injections of azoxymethane, once a week for 2 weeks. Seven days after the last injection, Wistar rats received fraction P (0.01%) dissolved in drinking water. After 6 weeks of treatment, the colon of rats receiving procyanidins showed a significant (P < 0.01) reduction of the number of preneoplastic lesions when compared with controls receiving water. The total number of hyperproliferative crypts and of aberrant crypt foci was reduced by 50% in rats receiving 0.01% apple procyanidins in their drinking water. Our results show that apple procyanidins alter intracellular signaling pathways, polyamine biosynthesis and trigger apoptosis in tumor cells. These compounds antagonize cancer promotion in vivo. In contrast with absorbable drugs, these natural, non toxic, dietary constituents reach the colon where they are able to exert their antitumor effects. PMID- 15790590 TI - Apoptosis and age-dependant induction of nuclear and mitochondrial etheno-DNA adducts in Long-Evans Cinnamon (LEC) rats: enhanced DNA damage by dietary curcumin upon copper accumulation. AB - Long-Evans Cinnamon (LEC) rats, a model for human Wilson's disease, develop chronic hepatitis and liver tumors owing to accumulation of copper and induced oxidative stress. Lipid peroxidation (LPO)-induced etheno-DNA adducts in nuclear- and mitochondrial-DNA along with apoptosis was measured in LEC rat liver. Levels of etheno-DNA adducts (1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine) increased with age reaching a peak at 8 and 12 weeks in nuclear and mitochondrial DNA, respectively. This is the first demonstration that etheno-DNA adducts are also formed in mitochondrial DNA. Apoptosis was assessed by TUNEL+ cells in liver sections. CD95L RNA expression was also measured by in situ hybridization in the same sections. The highest nuclear DNA adduct levels coincided with a reduced apoptotic rate at 8 weeks. Mitochondrial-DNA adducts peaked at 12 weeks that coincided with the highest apoptotic rate, suggesting a link of etheno-DNA adducts in mitochondrial DNA to apoptosis. The DNA damage in liver was further enhanced and sustained by 0.5% curcumin in the diet. Treatment for 2 weeks elevated etheno-DNA adducts 9- to 25-fold in nuclear DNA and 3- to 4-fold in mitochondrial-DNA, providing a plausible explanation as to why in our earlier study [Frank et al. (2003) Mutat. Res., 523-524, 127-135], curcumin failed to prevent liver tumors in LEC rats. Our results also confirm the reported in vitro DNA damaging potential of curcumin in the presence of copper ions by reactive oxygen species. LPO-induced adduct formation in nuclear and mitochondrial DNA appear as early lesions in LEC rat liver carcinogenesis and are discussed in relation to apoptotic events in the progression of malignant disease. PMID- 15790591 TI - Tobacco components stimulate Akt-dependent proliferation and NFkappaB-dependent survival in lung cancer cells. AB - Retrospective studies have shown that patients with tobacco-related cancers who continue to smoke after their diagnoses have lower response rates and shorter median survival compared with patients who stop smoking. To provide insight into the biologic basis for these clinical observations, we tested whether two tobacco components, nicotine or the tobacco-specific carcinogen, 4-(methylnitrosoamino)-1 (3-pyridyl)-1-butanone (NNK), could activate the Akt pathway and increase lung cancer cell proliferation and survival. Nicotine or NNK, rapidly and potently, activated Akt in non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC) cells. Nicotinic activation of Akt increased phosphorylation of multiple downstream substrates of Akt in a time-dependent manner, including GSK-3, FKHR, tuberin, mTOR and S6K1. Since nicotine or NNK bind to cell surface nicotinic acetylcholine receptors (nAchR), we used RT-PCR to assess expression of nine alpha and three beta nAchR subunits in five NSCLC cell lines and two types of primary lung epithelial cells. NSCLC cells express multiple nAchR subunits in a cell line-specific manner. Agonists of alpha3/alpha4 or alpha7 subunits activated Akt in a time-dependent manner, suggesting that tobacco components utilize these subunits to activate Akt. Cellular outcomes after nicotine or NNK administration were also assessed. Nicotine or NNK increased proliferation of NSCLC cells in an Akt-dependent manner that was closely linked with changes in cyclin D1 expression. Despite similar induction of proliferation, only nicotine decreased apoptosis caused by serum deprivation and/or chemotherapy. Protection conferred by nicotine was NFkappaB-dependent. Collectively, these results identify tobacco component-induced, Akt-dependent proliferation and NFkappaB-dependent survival as cellular processes that could underlie the detrimental effects of smoking in cancer patients. PMID- 15790592 TI - Identification of multiple nuclear export sequences in Fanconi anemia group A protein that contribute to CRM1-dependent nuclear export. AB - The Fanconi anemia (FA) pathway plays an important role in maintaining genomic stability, and defects in this pathway cause cancer susceptibility. The FA proteins have been found to function primarily in a nuclear complex, although a cytoplasmic localization and function for several FA proteins has also been reported. In this study, we investigated the possibility that FANCA, FANCC and FANCG are subjected to active export out of the nucleus. After treatment with leptomycin B, a specific inhibitor of CRM1-mediated nuclear export, the accumulation of epitope-tagged FANCA in the nucleus increased, whereas FANCC was affected to a lesser extent and FANCG showed no response. CRM1-mediated export of FANCA was further confirmed using CRM1 cotransfection, which led to a dramatic relocalization of FANCA to the cytoplasm. Five functional leucine-rich nuclear export sequences (NESs) distributed throughout the FANCA sequence were identified and characterized using an in vivo export assay. Simultaneous inactivation of three of these NESs resulted in a discrete but reproducible increase of FANCA nuclear accumulation. However, these NES mutations did not affect the ability of FANCA to complement the mitomycin C or cisplatin sensitivity of FA-A lymphoblasts. Surprisingly, mutations in the other two NESs resulted in an almost complete relocation of the protein to cytoplasm, suggesting that these motifs overlap with domains that are crucial for nuclear import. Taken together, these findings indicate that FANCA can be actively exported out of the nucleus by CRM1, revealing a new mechanism to regulate the function of the FA protein complex. PMID- 15790593 TI - Comparative evolutionary analysis of VPS33 homologues: genetic and functional insights. AB - VPS33B protein is a homologue of the yeast class C vacuolar protein sorting protein Vps33p that is involved in the biogenesis and function of vacuoles. Vps33p homologues contain a Sec1 domain and belong to the family of Sec1/Munc18 (SM) proteins that regulate fusion of membrane-bound organelles and interact with other vps proteins and also SNARE proteins that execute membrane fusion in all cells. We demonstrated recently that mutations in VPS33B cause ARC syndrome (MIM 208085), a lethal multisystem disease. In contrast, mutations in other Vps33p homologues result in different phenotypes, e.g. a mutation in Drosophila melanogaster car gene causes the carnation eye colour mutant and inactivation of mouse Vps33a causes buff hypopigmentation phenotype. In mammals two Vps33p homologues (e.g. VPS33A and VPS33B in humans) have been identified. As comparative genome analysis can provide novel insights into gene evolution and function, we performed nucleotide and protein sequence comparisons of Vps33 homologues in different species to define their inter-relationships and evolution. In silico analysis (a) identified two homologues of yeast Vps33p in the worm, fly, zebrafish, rodent and human genomes, (b) suggested that Carnation is an orthologue of VPS33A rather than VPS33B and (c) identified conserved candidate functional domains within VPS33B. We have shown previously that wild type VPS33B induced perinuclear clustering of late endosomes and lysosomes in human renal cells. Consistent with the predictions of comparative analysis: (a) VPS33B induced significantly more clustering than VPS33A in a renal cell line, (b) a putative fly VPS33B homologue but not Carnation protein also induced clustering and (c) the ability to induce clustering in renal cells was linked to two evolutionary conserved domains within VPS33B. One domain was present in VPS33B but not VPS33A homologues and the other was one of three regions predicted to form a t-SNARE binding site in VPS33B. In contrast, VPS33A induced significantly more clustering of melanosomes in melanoma cells than VPS33B. These investigations are consistent with the hypothesis that there are two functional classes of Vps33p homologues in all multicellular organisms and that the two classes reflect the evolution of organelle/tissue-specific functions. PMID- 15790594 TI - Association between a complex insertion/deletion polymorphism in NOD1 (CARD4) and susceptibility to inflammatory bowel disease. AB - The identification of the role of genetic variants within NOD2 (CARD15) in Crohn's disease and ulcerative colitis susceptibility highlight the role of the innate immune system in inflammatory bowel disease (IBD) pathogenesis. NOD1 (CARD4) is located on chromosome 7p14.3, in a region of known linkage to IBD and encodes an intracellular bacterial pathogen-associated molecular pattern receptor that is closely related to NOD2. We have identified strong association between haplotypes in the terminal exons of NOD1 and IBD (multi-allelic P = 0.0000003) in a panel of 556 IBD trios. The deletion allele of a complex functional NOD1 indel polymorphism (ND(1) + 32656*1) was significantly associated with early-onset IBD (P = 0.0003) in unrelated cases and controls. ND1 + 32656*1 was also associated with extra-intestinal manifestations of IBD (P = 0.04). These findings in two independent populations provide strong evidence for a role for NOD1 variants in IBD susceptibility and reinforce the role of the innate immune system in IBD pathogenesis. PMID- 15790595 TI - The Parkinson's disease-associated DJ-1 protein is a transcriptional co-activator that protects against neuronal apoptosis. AB - Mutations in the DJ-1 gene cause early-onset autosomal recessive Parkinson's disease (PD), although the role of DJ-1 in the degeneration of dopaminergic neurons is unresolved. Here we show that the major interacting-proteins with DJ-1 in dopaminergic neuronal cells are the nuclear proteins p54nrb and pyrimidine tract-binding protein-associated splicing factor (PSF), two multifunctional regulators of transcription and RNA metabolism. PD-associated DJ-1 mutants exhibit decreased nuclear distribution and increased mitochondrial localization, resulting in diminished co-localization with co-activator p54nrb and repressor PSF. Unlike pathogenic DJ-1 mutants, wild-type DJ-1 acts to inhibit the transcriptional silencing activity of the PSF. In addition, the transcriptional silencer PSF induces neuronal apoptosis, which can be reversed by wild-type DJ-1 but to a lesser extent by PD-associated DJ-1 mutants. DJ-1-specific small interfering RNA sensitizes cells to PSF-induced apoptosis. Both DJ-1 and p54nrb block oxidative stress and mutant alpha-synuclein-induced cell death. Thus, DJ-1 is a neuroprotective transcriptional co-activator that may act in concert with p54nrb and PSF to regulate the expression of a neuroprotective genetic program. Mutations that impair the transcriptional co-activator function of DJ-1 render dopaminergic neurons vulnerable to apoptosis and may contribute to the pathogenesis of PD. PMID- 15790596 TI - Sox9 is sufficient for functional testis development producing fertile male mice in the absence of Sry. AB - In the dominant mouse mutant Odd Sex, XXOds/+ mice develop as phenotypic, sterile males due to male-pattern expression of Sox9 in XXOds/+ embryonic gonads. To test whether SOX9 was sufficient to generate a fully fertile male in the absence of Sry, we constructed an XY(Sry(-))Ods/+ male mouse, in which the male phenotype is controlled autosomally by the Ods mutation. Mice were initially fertile, but progressively lost fertility until 5-6 months when they were sterile with very few germ cells in the testis. XY(Sry-)Ods/+ males also failed to establish the correct male-specific pattern of vascularization at the time of sex determination, which could be correlated to an inability of XY(Sry-),Ods/+ males to fully down-regulate Wnt4 expression in the embryonic gonad. Increasing the amount of SOX9 by producing homozygous XY(Sry-)Ods/Ods males was able to completely rescue the phenotype and restore correct vascular patterning and long term fertility. These data indicate that activation of SOX9 in the gonad is sufficient to trigger all the downstream events needed for the development of a fully fertile male and provide evidence that Sox9 may down-regulate Wnt4 expression in the gonad. PMID- 15790597 TI - Ethnic- and gender-specific association of the nicotinic acetylcholine receptor alpha4 subunit gene (CHRNA4) with nicotine dependence. AB - We tested six single nucleotide polymorphisms (SNPs) in the alpha4 subunit gene (CHRNA4) and four SNPs in the beta2 subunit gene (CHRNB2) of nicotinic acetylcholine receptors (nAChRs) for association with nicotine dependence (ND), which was assessed by smoking quantity (SQ), the heaviness of smoking index (HSI) and the Fagerstrom test for ND (FTND) in 2037 subjects from 602 nuclear families of either European-American (EA) or African-American (AA) ancestry. Analysis of the six SNPs within CHRNA4 demonstrated that in the EA sample SNPs rs2273504 and rs1044396 are significantly associated with the adjusted SQ and FTND score, respectively. In the AA samples, SNPs rs3787137 and rs2236196 are each significantly associated with at least two adjusted ND measures. Association of rs2236196 with the adjusted HSI and FTND scores in the AA samples remained significant after correction for multiple testing. Furthermore, analysis revealed gender- and ethnic-specific associations for several SNPs with ND measures in both ethnic samples; however, only the association of SNP rs2236196 with the three adjusted ND measures remained significant after correcting for multiple testing in the AA female samples. Haplotype analysis of rs2273505-rs2273504 rs2236196 showed significant association after Bonferroni correction of a C-G-G haplotype (53.4%) with three adjusted ND measures in samples from the AA females. A similar analysis for the four SNPs within CHRNB2 did not reveal significant association with the three ND measures. In summary, our findings provide convincing evidence for the involvement of the nAChR alpha4 subunit, but not of the nAChR beta2 subunit, in nicotine addiction. PMID- 15790598 TI - A non-sequence-specific requirement for SMN protein activity: the role of aminoglycosides in inducing elevated SMN protein levels. AB - Spinal muscular atrophy (SMA) is caused by homozygous loss of the survival motor neuron (SMN1) gene. In virtually all SMA patients, a nearly identical copy gene is present, SMN2. SMN2 cannot fully compensate for the loss of SMN1 because the majority of transcripts derived from SMN2 lack a critical exon (exon 7), resulting in a dysfunctional SMN protein. Therefore, the critical distinction between a functional and a dysfunctional SMN protein is the inclusion or the exclusion of the exon 7 encoded peptide. To determine the role of the 16 amino acids encoded by SMN exon 7, a panel of synthetic mutations were transiently expressed in SMA patient fibroblasts and HeLa cells. Consistent with previous reports, the protein encoded by SMN exons 1-6 was primarily restricted to the nucleus. However, a variety of heterologous sequences fused to the C-terminus of SMN exons 1-6 allowed mutant SMN proteins to properly distribute to the cytoplasm and to the nuclear gems. These data demonstrate that the SMN exon 7 sequence is not specifically required, rather this region functions as a non-specific 'tail' that facilitates proper localization. Therefore, a possible means to restore additional activity to the SMNDelta7 protein could be to induce a longer C terminus by suppressing recognition of the native stop codon. To address this possibility, aminoglycosides were examined for their ability to restore detectable levels of SMN protein in SMA patient fibroblasts. Aminoglycosides can suppress the accurate identification of translation termination codons in eukaryotic cells. Consistent with this, treatment of SMA patient fibroblasts with tobramycin and amikacin resulted in a quantitative increase in SMN-positive gems and an overall increase in detectable SMN protein. Taken together, this work describes the role of the critical exon 7 region and identifies a possible alternative approach for therapeutic intervention. PMID- 15790599 TI - Combined oral contraceptives in the treatment of polycystic ovary syndrome. AB - Combined oral contraceptives (COC) are the most often used treatment modality for polycystic ovary syndrome (PCOS). Undisputedly, COC suppress androgen production, thus ameliorating skin androgenic symptoms and improving menstrual dysfunction. On the other hand, there are still many unresolved issues concerning their metabolic effects. COC could decrease insulin sensitivity and deteriorate glucose tolerance, although the negative influence on insulin sensitivity is dependent on other factors (especially obesity) and this need not be expressed in non-obese patients. It is probable that the impairment of glucose tolerance is reversible, as the incidence of diabetes is not increased in past COC users. The effects of COC on the lipid spectrum are dependent on the type of gestagen, but lipid levels usually remain within the reference limits. Combination therapy of COC with weight reduction or insulin sensitizers could further suppress androgen levels and improve metabolic parameters. The establishment of COC after laparoscopic ovarian drilling may further decrease androgen levels. The combination of COC and GnRH analogues is not superior to COC therapy alone. Prospective data about the influence of COC on the risk of diabetes mellitus, coronary artery disease and endometrial cancer in PCOS women are lacking. PMID- 15790600 TI - Endocrine approaches in the therapy of prostate carcinoma. AB - At present, the management of non-organ confined prostate cancer, whether it is a recurrence or metastasis, continues to evolve based on prostate cancer detection using prostate-specific antigen and the development of medications as alternatives for the classical orchiectomy, which induced irreversible implications for quality of life. Diethylstilbestrol therapy was associated with cardiovascular side-effects; GnRH agonists were able to create a castration level, but again considerable side-effects were described. Combination therapies using antiandrogens and GnRH agonists do not improve survival and have additional toxicity. GnRH antagonists, which also suppress FSH, represent the latest class of agents introduced for hormonal treatment, but phase III studies with survival data are not yet available. In spite of all these achievements, hormonal manipulation has resulted in only modest improvements during recent decades and new targets are needed to improve the clinical outcome. Selectively modifying the androgen receptor is currently one of the most promising developments. PMID- 15790601 TI - The role of paf in embryo physiology. AB - Embryo-derived paf (1-o-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is produced by de novo synthesis. This synthesis commences soon after fertilization and persists throughout the preimplantation phase. Paf is produced and released by the embryos of all mammalian species studied to date. Its release from the embryo involves binding to extracellular albumin in a manner that protects paf from enzymatic degradation. Released paf causes a range of alterations in maternal physiology, including platelet activation, changes in oviductal, endometrial and maternal immune function. Paf also acts in an autocrine fashion as a trophic/survival factor for the early embryo. In vitro, supplementation of culture media with paf improves embryo development. Embryo-derived paf's autocrine actions are transduced by 1-o-phosphatidylinositol-3-kinase, which induces characteristic calcium transients within the early embryo. The calcium transients require both the influx of external calcium and release of inositol trisphosphate-dependent internal calcium stores. Buffering these transients compromised embryo development in a manner that was reversed by exogenous paf. Assisted reproductive technologies compromise the production of paf by some embryos and retard the expression of the paf receptor. This deprivation of paf's action is one of the factors limiting the survivability of embryos produced by assisted reproductive technologies. Paf is one of several autocrine and paracrine trophic/survival factors that act on the early embryo. These factors probably act cooperatively and may, to some degree, be mutually redundant. As the earliest released and the best-described embryotrophin, paf provides an important exemplar for understanding the role of ligand-mediated trophic support of the early embryo. PMID- 15790602 TI - Selective progesterone receptor modulators and progesterone antagonists: mechanisms of action and clinical applications. AB - Since the discovery of the antiprogestin mifepristone, hundreds of similar compounds have been synthesized, which can be grouped in a large family of progesterone receptor ligands. This family includes pure agonists such as progesterone itself or progestins and, at the other end of the biological spectrum, pure progesterone receptor antagonists (PA). Selective progesterone receptor modulators (SPRM) have mixed agonist-antagonist properties, and occupy an intermediate position of the spectrum. These compounds have numerous applications in female health care. Mifepristone is used to terminate pregnancy, and as such is commercially available in many countries. The negative abortion related image of mifepristone has clearly limited the involvement of the major pharmaceutical companies in the development of PA and SPRM. Many PA and SPRM display direct antiproliferative effects in the endometrium, although with variable actions which seem product- and dose-dependent. This property justifies their use in the treatment of myomas and endometriosis. PA also suppress late follicular development, block the LH surge and retard endometrial maturation, which renders them potential estrogen-free contraceptive drugs. SPRM such as asoprisnil are not as effective in blocking the LH surge and appear to target the endometrium directly and produce amenorrhoea. Interestingly, clinical data show that treatment with these compounds is not associated with hypo-estrogenism and bone loss. The potential clinical applications of these compounds cover a broad field and are very promising in major public health areas. These include emergency contraception, long-term estrogen-free contraception (administered alone, or in association with a progestin-only pill to improve bleeding patterns), myomas (where they induce a marked reduction in tumour volume and produce amenorrhoea) and endometriosis. Further developments might also include hormone replacement therapy in post-menopausal women, as well as the treatment of hormone-dependent tumours. PMID- 15790603 TI - Life table (survival) analysis to generate cumulative pregnancy rates in assisted reproduction: are we overestimating our success rates? AB - The variability in the numbers of treatment cycles couples may undertake with assisted reproductive technology (ART) and the length of time they may have to wait between successive cycles of treatment make the evaluation of treatment efficacy and prognosis complicated. The cumulative pregnancy rate using the life table method of analysis is being used more frequently to estimate the effectiveness of treatment. Although this approach is valid in some areas of infertility research, its use in ART is not appropriate, because the factors necessary for the analysis (particularly the scale for measuring the passage of time and lack of informative censoring) are not satisfied. Consequently, an overestimation of the effect of treatment is produced that may lead to biased decision making. Although there is no easy solution to this problem, several options for summarizing the outcome data are offered: pregnancy rate per cycle, time-limited analysis using proportions, conservative cycle-based cumulative pregnancy rate and real-time-based cumulative pregnancy rate. In this manner, more realistic information can be generated to counsel patients, evaluate the efficacy of treatments, compare rates among centres and guide the formulation of policies for infertility management and resource allocation. PMID- 15790604 TI - Observational clinical follow-up of oocyte cryopreservation using a slow-freezing method with 1,2-propanediol plus sucrose followed by ICSI. AB - BACKGROUND: The value of oocyte cryopreservation remains controversial. Two major problems exist: poor survival and injury to the oocyte meiotic spindle after freezing and thawing. METHODS: For slow oocyte cryopreservation, we used 1.5 mol/l 1,2-propanediol and 0.3 mol/l sucrose. We waited 3 h after thawing for possible recovery of the meiotic spindles before performing ICSI. RESULTS: Forty three women undergoing IVF or ICSI cycles cryopreserved some or all of their harvested oocytes; of these, 20 thawed their cryopreserved oocytes for personal use and one for donation. The survival rate of oocytes after thawing was 75%, with 67% of oocytes fertilizing normally after ICSI. All 21 cycles (100%) resulted in fertilization and embryo transfers. Seven pregnancies (33%) resulted. Four women delivered five babies with normal karyotypes. Three conceptions are ongoing. Compared to 38 cycles of frozen-thawed embryos at the pronuclear stage in the same period, the percentages of survival, pregnancy and implantation were similar. Additionally, four unmarried women with white blood cell diseases underwent oocyte freezing before preconditioning treatment for haematopoietic stem cell transplantation. CONCLUSIONS: This protocol achieved reproducible success of survival, fertilization and pregnancy for freezing and thawing of human oocytes. The 3 h post-thaw incubation could permit restoration of the meiotic spindles, thus facilitating normal fertilization. PMID- 15790605 TI - Relationships in couples after failed IVF treatment: a prospective follow-up study. AB - BACKGROUND: There are few studies of couples that analyse satisfaction with treatment, adoption plans and relationships in couples after unsuccessful IVF. METHODS: ENRICH marital inventory was used to describe marital dynamics and to gain information about treatment and adoption plans. A specially designed questionnaire was used. Of the 51 couples without previous children who were asked to participate after their first failed IVF cycle, 45 participated. The next stage of the study was carried out when the couples had reached the 6 months point after the first IVF cycle, and the last stage after the couples had been through one to three treatments, 1(1/2) years after the last treatment. RESULTS: The couples displayed a stable relationship from the start as well as 1 year after the last IVF cycle. The vast majority of the couples had decided to go through with an adoption. Seventy-three per cent of the women were interested in more IVF treatment compared to 33% of the men. CONCLUSION: The stresses associated with IVF treatment did not have a negative impact on the couples' appreciation of their relationships during and after the treatment period. After treatment had been completed, the couples seemed to have reoriented themselves toward other solutions to childlessness. PMID- 15790606 TI - Italy enacts new law on medically assisted reproduction. AB - In 2004, the Italian Parliament enacted a law regulating medically assisted reproduction. Although the law recognizes as legal certain assisted reproduction techniques, several other procedures are implicitly or expressly banned: oocyte and sperm donation, using embryos for the scientific research purposes and reproductive cloning. In this article, I outline the new legal framework, pointing out some of the shortcomings of its provisions, such as the failure to define what an 'embryo' is, the contradictions between this law and the law on abortion, the opportunity for Italian couples to circumvent some of the prohibitions by resorting to 'reproductive tourism', and the central role that physicians play in the new legal framework. PMID- 15790607 TI - Randomized clinical trial of a levonorgestrel-releasing intrauterine system and a depot GnRH analogue for the treatment of chronic pelvic pain in women with endometriosis. AB - BACKGROUND: The objective of this multicentre randomized, controlled clinical trial was to compare the efficacy of a levonorgestrel-releasing intrauterine system (LNG-IUS) and a depot-GnRH-analogue in the control of endometriosis related pain over a period of six months. METHODS: Eighty-two women, 18 to 40 years of age (mean 30 years), with endometriosis, dysmenorrhoea and/or CPP, were randomized using a computer-generated system of sealed envelopes into either LNG IUS (n = 39) or GnRH analogue (n = 43) treatment groups at three university centres. Daily scores of endometriosis-associated CPP were evaluated using the Visual Analogue Scale (VAS), daily bleeding score was calculated from bleeding calendars, and improvement in quality of life was evaluated using the Psychological General Well-Being Index Questionnaire (PGWBI). The pain score diary was based on the VAS in which women recorded the occurrence and intensity of pain on a daily basis. A monthly score was calculated from the result of the sum of the daily scores divided by the number of days in each observation period. RESULTS: CPP decreased significantly from the first month throughout the six months of therapy with both forms of treatment and there was no difference between the groups (P > 0.999). In both treatment groups, women with stage III and IV endometriosis showed a more rapid improvement in the VAS pain score than women with stage I and II of the disease (P < 0.002). LNG-IUS users had a higher bleeding score than GnRH-analogue users at all time points of observation with 34% and 71% of patients in the LNG-IUS and GnRH-analogue groups, respectively, reporting no bleeding during the first treatment month, and 70% and 98% reporting no bleeding during the sixth month. No difference was observed between groups with reference to improvement in quality of life. CONCLUSIONS: Both, the LNG-IUS and the GnRH-analogue were effective in the treatment of CPP-associated endometriosis, although no differences were observed between the two treatments. Among the additional advantages of the LNG-IUS is the fact that it does not provoke hypoestrogenism and that it requires only one medical intervention for its introduction every 5 years. This device could therefore become the treatment of choice for CPP-associated endometriosis in women who do not wish to conceive. PMID- 15790608 TI - In vivo gene transfer of lefty leads to implantation failure in mice. AB - BACKGROUND: Endometrium is a unique tissue that is prepared for implantation of blastocyst during each menstrual cycle. In humans, if implantation does not occur or fails, endometrium is shed. METHODS AND RESULTS: We identified ebaf/lefty, as a key cytokine, highly expressed in human endometrium during the non-receptive phase of tissue remodelling. Lefty was increased in the endometria of a number of patients with 'unexplained infertility' during the receptive phase, suggesting dysregulation of lefty as a potential factor contributing to infertility. Here, we showed that induction of a similar state of lefty overexpression in endometrium, by in vivo gene delivery, decreased implantation in pregnant mice. This state of overexpression could be induced by a retroviral vector transducing lefty or by liposome-mediated introduction of a lefty expression vector. Analysis of endometrium showed increased lefty after in vivo gene transfer. CONCLUSION: These findings suggest that induction of a state of lefty overexpression in endometrium leads to reduced implantation. PMID- 15790609 TI - The choice and outcome of the fertility treatment of 38 couples in whom the male partner has a Yq microdeletion. AB - BACKGROUND: Patients with Yq microdeletions may suffer from fertility problems. The purpose of this study was to assess the outcome of the fertility treatment of these patients. METHODS: For 38 patients with Yq microdeletions, data were collected about medical history, karyotype, testicular histopathology and the presence of spermatozoa in the ejaculate or testicular biopsies. RESULTS: Sixteen patients with an azoospermia factor region c (AZFc) deletion had at least one cycle with ICSI. The clinical pregnancy outcome was 22% per embryo transfer. Three babies have been born and two pregnancies are ongoing. Sex selection for female embryos in combination with ICSI in order to prevent the transmission of the fertility problems was discussed with 14 couples; eight were in favour of the selection. In addition, eight couples made the decision to use donor sperm because ICSI was impossible or objectionable and, in this group of patients, a total of nine babies were born. CONCLUSIONS: Despite the improvement of assisted reproductive technology, ICSI could be offered only to patients with an AZFc deletion. Insemination with donor sperm is a potential alternative for other patients. PMID- 15790610 TI - Administration of B-group vitamins reduces circulating homocysteine in polycystic ovarian syndrome patients treated with metformin: a randomized trial. AB - BACKGROUND: The aim of the current study was to assess the effects of B-group vitamins and folic acid administration on serum levels of homocysteine (Hcy) in patients with polycystic ovarian syndrome (PCOS) on short-term metformin treatment. METHODS: Patients were randomly assigned to one of three treatment groups. Group 1 patients (n = 20) received metformin (850 mg twice daily); group 2 patients (n = 20) received metformin (850 mg twice daily) and B-group vitamins (vitamin B1, 250 mg; vitamin B6, 250 mg; vitamin B12, 1000 microg twice daily); and group 3 patients (n = 20) received metformin (850 mg twice daily) and folic acid (174 microg twice daily). In all groups, lipid profiles and plasma total Hcy, vitamin B12, folic acid and glucose levels were recorded at baseline and at 3 months. RESULTS: A 26.5% increase in Hcy levels was seen after 12 weeks of metformin therapy, while 21.17 and 8.33% decreases in Hcy levels were detected when B-group vitamins or folic acid plus metformin were given respectively. There were no statistically significant differences recorded in insulin sensitivity using homeostasis model assessment in the three groups. CONCLUSION: These findings suggest that B-group vitamins and folic acid administration counteract the Hcy-increasing effect seen with metformin therapy. PMID- 15790611 TI - Comparison of outcome of pregnancy after intra-uterine insemination (IUI) and IVF. AB - INTRODUCTION: Pregnancy outcome after IVF has been shown to be worse than after spontaneous conception. There is discussion as to whether this results from the technique itself or the patient characteristics. This study compares pregnancy outcome after IVF and intra-uterine inemination (IUI) in a matched patient group. METHODS: Data were obtained from our IVF and IUI databases (1997-2001). Matching was performed for maternal age, parity and plurality, and 126 IUI pregnancies were compared with 126 IVF pregnancies. Outcome variables were pregnancy duration, birth weight, Caesarean section rates, preterm contraction rates, neonatal intensive care unit admission, Apgar score, blood loss rates and maternal hypertension. RESULTS: None of the analysed parameters was statistically different between the groups. CONCLUSION: This matched case-control study does not show different pregnancy outcomes after IVF and IUI. Since there is no reason to believe that the IUI technique in itself leads to an increased obstetric or neonatal risk, this study suggests that the worse pregnancy outcome after IVF as compared with spontaneous conceptions is due to the specific patient characteristics, rather than to the use of IVF itself. PMID- 15790612 TI - Relationship between urinary estrogen levels before conception and sex ratio at birth in a primate, the gray mouse lemur. AB - BACKGROUND: In primates, including humans, bias of sex ratio at birth has been reported to depend on maternal condition at conception. In a Malagasy primate, the gray mouse lemur, male-biased sex ratio at birth occurred in captive parous females. The male bias was previously demonstrated to be pre-conceptual and independent of the female's nutritional state. It was hypothesized to be related to changes in hormonal state at ovulation. METHODS AND RESULTS: The relationship between urinary estradiol (E2) levels during the follicular phase until estrus and litter production (number and sex of newborns) was assessed in 91 females mated under controlled conditions. Changes in E2 levels prior to ovulation followed the typical primate pattern characterized by a gradual rise during the 10 days preceding the sharp increase at estrus. A clear decline in E2 levels occurred with ageing. Direction of the sex ratio bias was unrelated to E2 levels at ovulation time but was significantly dependent on E2 levels during the follicular phase. Reduced E2 levels prior to estrus led to male-biased litters. CONCLUSION: This study suggests that hormonal stimulation during the follicular phase plays a role in shifting sex ratio at conception through changes in the local environment of the ova. This hypothesis deserves testing by assessing estrogen levels throughout the follicular phase in other primate species including humans. PMID- 15790613 TI - Titanium dioxide: inhalation toxicology and epidemiology. AB - Titanium dioxide (TiO(2)) is manufactured worldwide in large quantities for use in a wide range of applications and is normally considered to be toxicologically inert. Findings of tumours in the lungs of rats exposed chronically to high concentrations of TiO(2), but not in similarly exposed mice or hamsters, suggest that the tumorigenic response may be a rat-specific phenomenon but nonetheless raises concerns for potential human health effects. With the limited toxicological understanding of species differences in response to inhaled TiO(2) and a similarly limited amount of epidemiological information with respect to TiO(2) exposure in the workplace, a consortium of TiO(2) manufacturers in Europe (under the European Chemistry Industry Council; CEFIC) and in North America (under the American Chemistry Council; ACC) initiated a programme of research to investigate inter-species differences as a result of exposure to TiO(2) and to conduct detailed epidemiological surveys of the major manufacturing sites. The toxicology studies exposed rats, mice and hamsters to pigment-grade TiO(2) (PG TiO(2), 0, 10, 50 and 250 mg m(-3)) or ultrafine TiO(2) (UF-TiO(2), 0, 0.5, 2 and 10 mg m(-3)) for 90 days and the lung burdens and tissue responses were evaluated at the end of the exposure period and for up to 1 year after exposure. Results demonstrated clear species differences. Rats and mice had similar lung burdens and clearance rates while hamsters showed high clearance rates. At high lung particle burdens, rats showed a marked progression of histopathological lesions throughout the post-exposure period while mice and hamsters showed minimal initial lesions with recovery apparent during the post-exposure period. Lung neutrophil responses, a sensitive marker of inflammatory changes, reflected the development or recovery of the histopathological lesions. The use of surface area rather than gravimetric lung burden provided closer correlates of the burden to the biological effect across both TiO(2) types. The epidemiological investigations evaluated the mortality statistics at 11 European and 4 US TiO(2) manufacturing plants. They concluded that there was no suggestion of any carcinogenic effect associated with workplace exposure to TiO(2). PMID- 15790614 TI - An experimental study to investigate the feasibility to classify paints according to neurotoxicological risks: occupational air requirement (OAR) and indoor use of alkyd paints. AB - The concept of occupational air requirement (OAR), representing the quantity of air required to dilute the vapor concentration in the work environment resulting from 1 l product to a concentration below the occupational exposure limit (OEL), was considered to have potential to discriminate between paints that can and cannot be used safely. The OAR is a simple algorithm with the concentration of volatile organic compound (VOC) in the paint, a discrete evaporation factor and the neurotoxicological effects-based OEL. Conceptually, OAR categories of paints for construction and maintenance applications could be identified that can be applied manually without exceeding OELs with no appreciable room ventilation. Five painters volunteered in an exposure study aimed at testing the OAR approach in practice. Total exposure to VOC was assessed in 30 experiments during the application of 0.5 l of paint in a defined 'standard indoor paint job'. Fifteen paints were prepared, reflecting differences in solvents (percentage, volatility, toxicity) with a range of OAR levels from 43 to 819 m(3)/l. Exposure was assessed by personal air sampling (PAS). In addition, real-time air monitoring was performed. All tests were conducted at minimum ventilation rate (< or=0.33 h( 1)). PAS results were expressed as percentage of the nominal OEL and ranged from 8 to 93% for high solids and from 38 to 168% for conventional paints. In general, higher VOC contents resulted in higher exposure. High volatile paints showed a statistically significant faster increase of VOC concentration with time compared with paints containing low volatile solvents. A significant relationship between OAR value and exposure was observed (R(2) = 0.73). The experiments indicate that OAR-based classification of paints predicts and discriminates risk levels for exposure to neurotoxic paint-solvents in indoor painting fairly well. PMID- 15790615 TI - Thoracic size-selective sampling of fibres: performance of four types of thoracic sampler in laboratory tests. AB - The counting of fibres on membrane filters could be facilitated by using size selective samplers to exclude coarse particulate and fibres that impede fibre counting. Furthermore, the use of thoracic size selection would also remove the present requirement to discriminate fibres by diameter during counting. However, before thoracic samplers become acceptable for sampling fibres, their performance with fibres needs to be determined. This study examines the performance of four thoracic samplers: the GK2.69 cyclone, a Modified SIMPEDS cyclone, the CATHIA sampler (inertial separation) and the IOM thoracic sampler (porous foam pre selector). The uniformity of sample deposit on the filter samples, which is important when counts are taken on random fields, was examined with two sizes of spherical particles (1 and 10 microm) and a glass fibre aerosol with fibres spanning the aerodynamic size range of the thoracic convention. Counts by optical microscopy examined fields on a set scanning pattern. Hotspots of deposition were detected for one of the thoracic samplers (Modified SIMPEDS with the 10 microm particles and the fibres). These hotspots were attributed to the inertial flow pattern near the port from the cyclone pre-separator. For the other three thoracic samplers, the distribution was similar to that on a cowled sampler, the current standard sampler for fibres. Aerodynamic selection was examined by comparing fibre concentration on thoracic samples with those measured on semi isokinetic samples, using fibre size (and hence calculated aerodynamic diameter) and number data obtained by scanning electron microscope evaluation in four laboratories. The size-selection characteristics of three thoracic samplers (GK2.69, Modified SIMPEDS and CATHIA) appeared very similar to the thoracic convention; there was a slight oversampling (relative to the convention) for d(ae) < 7 microm, but that would not be disadvantageous for comparability with the cowled sampler. Only the IOM thoracic sampler tended to undersample the fibres relative to the thoracic convention. With the data divided into four classes based on fibre length, the size-selection characteristics appeared to be unaffected by fibre length for GK2.69, Modified SIMPEDS and CATHIA. Only the IOM thoracic sampler (with the foam selector) showed slightly lower selection for longer length classes of fibres. These results indicate that the tested samplers follow the thoracic sampling convention for fibres, and may be used to improve the quality and reliability of samples that are taken when there is likely to be significant background dust. PMID- 15790616 TI - Statin use in the secondary prevention of coronary heart disease in primary care: cohort study and comparison of inclusion and outcome with patients in randomised trials. AB - OBJECTIVE: To compare the social and demographic profiles of patients who receive statin treatment after myocardial infarction and patients included in randomised trials. To estimate the effect of statin use in community based patients on subsequent all cause mortality and cardiovascular recurrence, contrasting effects with trial patients. DESIGN: Observational cohort study using a record linkage database. SETTING: Tayside, Scotland (population size and characteristics: about 400,000, mixed urban and rural). SUBJECTS: 4892 patients were discharged from hospital after their first myocardial infarction between January 1993 and December 2001. 2463 (50.3%) were taking statins during an average follow-up of 3.7 years (3.1% in 1993 and 62.9% in 2001). MAIN OUTCOME MEASURES: All cause mortality and recurrence of cardiovascular events. RESULTS: 319 deaths occurred in the statin treated group (age adjusted rate 4.1 per 100 person years, 95% confidence interval 3.2 to 4.9), and 1200 in the statin untreated group (12.7 per 100 person years, 11.1 to 14.3). More older people and women were represented in the population of patients treated with statins than among those recruited into clinical trials (mean age 67.8 v 59.8; women 39.6% v 16.9%, respectively). The effects of statins in routine clinical practice were consistent with, and similar to, those reported in clinical trials (adjusted hazard ratio for all cause mortality 0.69, 95% confidence interval 0.59 to 0.80; adjusted hazard ratio for cardiovascular recurrence 0.82, 0.71 to 0.95). CONCLUSIONS: The community effectiveness of statins in those groups that were not well represented in clinical trials was similar to the efficacy of statins in these trials. PMID- 15790617 TI - Highly active antiretroviral therapy. PMID- 15790618 TI - Nurse led care. PMID- 15790619 TI - Large scale food retail interventions and diet. PMID- 15790620 TI - A theme issue "by, for, and about" Africa. PMID- 15790621 TI - New standards for cardiopulmonary resuscitation. PMID- 15790622 TI - President Bush and Congress intervene in "right to die" case. PMID- 15790623 TI - Government launches plan to overhaul social care services. PMID- 15790624 TI - Doctors who performed late abortion will not be prosecuted. PMID- 15790625 TI - HIV treatment should be free for everyone in Britain, MPs say. PMID- 15790626 TI - Psychological therapy is better than debriefing for PTSD. PMID- 15790628 TI - German scientists demand a ban on tobacco additives. PMID- 15790629 TI - Israel tightens up hospital research procedures. PMID- 15790631 TI - US campaigners challenge abortion law changes. PMID- 15790634 TI - New cases of asthma in children in England and Wales has fallen by two thirds. PMID- 15790636 TI - Private providers must be stopped from skimming off easy cases. PMID- 15790637 TI - Researcher to be sacked after reporting high rates of ADHD. PMID- 15790638 TI - Government will not back down over free personal care for all elderly people. PMID- 15790639 TI - Falling EU population will affect healthcare provision. PMID- 15790640 TI - Changes to India's patent law may deny cheap drugs to millions. PMID- 15790642 TI - Reassuring evidence on competency based selection. PMID- 15790641 TI - A new selection system to recruit general practice registrars: preliminary findings from a validation study. AB - OBJECTIVE: To design and validate a new competency based selection system to recruit general practice registrars, comprising a competency based application form, referees' reports, and an assessment centre. DESIGN: Longitudinal predictive validity study and a matched case comparison. SETTING: South Yorkshire and East Midlands region, United Kingdom, comprising three deaneries. PARTICIPANTS: 46 of 167 doctors were followed up in training after three months in practice, and 20 general practice trainers were selected by using traditional recruitment methods. MAIN OUTCOME MEASURES: Trainer ratings of trainee performance in practice on targeted competencies. RESULTS: Performance ratings of targeted competencies at the assessment centre predicted trainer ratings of performance in the job. Furthermore, those trainees recruited through the new competency based process performed significantly better in the job than those recruited through traditional recruitment processes. CONCLUSION: A new competency based selection process using assessment centres improves the validity of selection of general practice registrars compared with traditional selection techniques. PMID- 15790644 TI - Recent developments in the use of biologics in psoriasis and autoimmune disorders. The role of autoantibodies. PMID- 15790645 TI - Chronic illness and disability. PMID- 15790646 TI - Evidence based diagnostics. PMID- 15790647 TI - Why is the grass greener? PMID- 15790648 TI - Authors of TADS study reply to letter raising concerns. PMID- 15790649 TI - Family history of breast cancer: referral guidelines changed after acceptance of 10 minute consultation. PMID- 15790650 TI - Recent advances in non-invasive cardiology: coronary angiography using computed tomography has been underplayed. PMID- 15790651 TI - Recent advances in non-invasive cardiology: article does not mention echocardiography. PMID- 15790652 TI - Alcohol in the body: elimination of alcohol from blood varies. PMID- 15790653 TI - Four pillars of academic medicine need to be restored. PMID- 15790654 TI - Deficiencies in disaster funding: malaria epidemics are predicted in tsunami regions from El Nino conditions. PMID- 15790655 TI - Deficiencies in disaster funding: disasters less "telegenic" than the tsunami must not be forgotten. PMID- 15790656 TI - Primary care research networks may herald new collaborations. PMID- 15790657 TI - How do developing synapses acquire AMPA receptors? PMID- 15790658 TI - Molecular, cellular and endocrine signalling in the perinatal cardiovascular system: interplay and developmental programming. PMID- 15790659 TI - Lessons from the leptin paradox in cardiac regulation--too much versus too little. PMID- 15790660 TI - Null mutation of myeloperoxidase in mice prevents mechanical activation of neutrophil lysis of muscle cell membranes in vitro and in vivo. AB - Membrane lysis is a common and early defect in muscles experiencing acute injuries or inflammation. Although increased mechanical loading of muscles can induce inflammation and membrane lysis, whether mechanical loads applied to muscle can promote the activation and cytolytic capacity of inflammatory cells and thereby increase muscle damage is unknown. We tested whether mechanical loads applied to mouse muscle cells in vitro can increase membrane lysis, and whether neutrophil-mediated lysis of muscle cells is promoted by mechanical loads applied in vitro and in vivo. Cyclic loads applied to muscle cells for 24 h in vitro produced little muscle cell lysis. Similarly, the addition of neutrophils to muscle cell cultures in the presence of superoxide dismutase (SOD) produced little muscle cell lysis. However, when cyclic mechanical loads were applied to neutrophil-muscle co-cultures in the presence of SOD, there was a synergistic effect on muscle cell lysis, suggesting that mechanical loading activates neutrophil cytotoxicity. However, application of mechanical loads to co-cultures of muscle cells and neutrophils that are null mutants for myeloperoxidase (MPO) showed no mechanical activation of neutrophil cytotoxicity. This indicates that loading promotes neutrophil cytotoxicity via MPO. Activity assays confirmed that mechanical loading of neutrophil-muscle co-cultures significantly increased MPO activity. We further tested whether muscle membrane lysis in vivo was mediated by neutrophils when muscle was subjected to modified loading by using a mouse model of muscle reloading following a period of unloading. We observed that MPO-/ soleus muscles showed a significant 52% reduction in membrane lysis compared to wild-type mice, although the mutation did not decrease inflammatory cell extravasation. Together, these in vitro and in vivo findings show that mechanical loading activates neutrophil-mediated lysis of muscle cells through an MPO dependent pathway. PMID- 15790661 TI - Reversals of anticipatory postural adjustments during voluntary sway in humans. AB - We describe reversals of anticipatory postural adjustments (APAs) with the phase of a voluntary cyclic whole-body sway movement. Subjects (n=9) held a standard load in extended arms and released it by a bilateral shoulder abduction motion in a self-paced manner at different phases of the sway. The load release task was also performed during quiet stance in three positions: in the middle of the sway range and close to its extreme forward and backward positions. Larger APAs were seen during the sway task as compared to quiet stance. Although the direction of postural perturbation associated with the load release was always the same, the direction of the APAs in the leg muscles reversed when the subjects were close to the extreme forward position as compared to the APAs in other phases and during quiet stance. The trunk muscles showed smaller APA modulation at the extreme positions but larger modulation when passing through the middle position, depending on the direction of sway, forward or backward. The phenomenon of APA reversals emphasizes the important role of safety in the generation of postural adjustments associated with voluntary movements. Based on these findings, APAs could be defined as changes in the activity of postural muscles associated with a predictable perturbation that act to provide maximal safety of the postural task component. PMID- 15790662 TI - beta-Adrenoceptor-mediated long-term up-regulation of the release machinery at rat cerebellar GABAergic synapses. AB - Properly regulated interactions among excitatory and inhibitory synapses are critical for brain function. Compared to excitatory synapses, much less is known about the gain control mechanisms at inhibitory synapses. Herein we report a mechanism of noradrenergic long-term potentiation (LTP) at inhibitory synapses following presynaptic beta-adrenoceptor activation. Stimulation of beta adrenoceptors elicited LTP of GABA release from terminals of cerebellar interneurones. This action was dependent on the cAMP/protein kinase A signalling cascade and independent of the beta-adrenoceptor-mediated acceleration of hyperpolarization-activated cyclic nucleotide-gated cation (HCN) channel. Furthermore, the beta-adrenoceptor- and protein kinase A-mediated LTP was triggered by enhancement of the Ca2+ sensitivity of the release machinery and increase in the readily releasable pool. beta-Adrenoceptor activation also accelerated the recruitment of GABA into the releasable pool and enhanced synchronous and asynchronous release of GABA from the presynaptic terminal. Thus, the up-regulation of GABA release machinery mediated by noradrenaline and beta adrenoceptor activation provides a likely mechanism of feedforward inhibition of the cerebellar output neurone Purkinje cell, leading to a profound effect on motor control and learning associated with the cerebellum. PMID- 15790663 TI - Maternal nutrient restriction in sheep: hypertension and decreased nephron number in offspring at 9 months of age. AB - Pregnant ewes were fed either a 50% nutrient-restricted (NR; n= 8) or a control 100% (C; n= 8) diet from day 28 to day 78 of gestation (dGA; term = 150 dGA). Lambs were born naturally, and fed to appetite throughout the study period. At 245 +/- 1 days postnatal age (DPNA), offspring were instrumented for blood pressure measurements, with tissue collection at 270 DPNA. Protein expression was assessed using Western blot, glomerulus number determined via acid maceration and hormone changes by radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). NR lambs had higher mean arterial pressure (MAP; 89.0 +/- 6.6 versus 73.4 +/- 1.6 mmHg; P < 0.05), fewer renal glomeruli (57.8 +/- 23.8 versus 64.6 +/ 19.3 x 10(4); P < 0.05), increased expression of angiotensin converting enzyme (ACE) in the renal cortex (942 +/- 130 versus 464 +/- 60 arbitrary pixel units (apu); P < 0.03), and increased angiotensin II receptor AT2 expression in the renal medulla (63.3 +/- 12.1 versus 19.5 +/- 44.2 x 10(4) apu; P < 0.03). All data are presented as mean +/-S.E.M. The present data indicate that global maternal nutrient restriction (50%) during early to mid-gestation impairs renal nephrogenesis, increases MAP, and alters expression of AT2 and ACE without an associated change in birth weight. These data demonstrate the existence of a critical window of fetal susceptibility during early to mid-gestation that alters kidney development and blood pressure regulation in later life. PMID- 15790664 TI - Occludin as direct target for glucocorticoid-induced improvement of blood-brain barrier properties in a murine in vitro system. AB - Homeostasis of the central nervous system (CNS) microenvironment is essential for its normal function. It is maintained by the blood-brain barrier (BBB) which regulates the transport of molecules from blood into brain and backwards. The integrity of the BBB is compromised in many disorders of the human CNS; therapeutical strategies for several of these diseases include treatment with glucocorticoids, but the molecular basis of how glucocorticoids regulate BBB permeability is not understood. Here, we report the generation and characterization of a murine immortalized brain (cerebral) capillary endothelial (cEND) cell line which expresses the BBB marker occludin at intercellular tight junctions (TJ). Hydrocortisone at physiological concentrations induced upregulation of occludin, accompanied by a threefold enhancement of transendothelial electrical resistance to values up to 1000 Omegacm2. Insulin enhanced the glucocorticoid response. At the molecular level, hydrocortisone induces increase of occludin at protein and mRNA levels by activation of the glucocorticoid receptor (GR) and its binding to putative glucocorticoid responsive elements in the occludin promoter. At the same time, insulin potentiated the ligand-dependent GR transactivation via induction of the GR in this in vitro system. This study thus provides insights into the molecular processes of barrier genesis, and may help to elucidate mechanisms of brain pathology at the microvascular level. PMID- 15790665 TI - Central command and the cutaneous vascular response to isometric exercise in heated humans. AB - Cutaneous vascular conductance (CVC) decreases during isometric handgrip exercise in heat stressed individuals, and we hypothesized that central command is involved in this response. Seven subjects performed 2 min of isometric handgrip exercise (35% of maximal voluntary contraction) followed by postexercise ischaemia in normothermia and during heat stress (increase in internal temperature approximately 1 degrees C). To augment the contribution of central command independent of force generation, on a separate day the protocol was repeated following partial neuromuscular blockade (PNB; i.v. cisatracurium). Forearm skin blood flow was measured by laser-Doppler flowmetry, and CVC was the ratio of skin blood flow to mean arterial pressure. The PNB attenuated force production despite encouragement to attain the same workload. During the heat stress trials, isometric exercise decreased CVC by approximately 12% for both conditions, but did not change CVC in either of the normothermic trials. During isometric exercise in the heat, the increase in mean arterial pressure (MAP) was greater during the control trial relative to the PNB trial (31.0+/-9.8 versus 18.6+/-6.4 mmHg, P<0.01), while the elevation of heart rate tended to be lower (19.4+/-10.4 versus 27.4+/-8.1 b.p.m., P=0.15). During postexercise ischaemia, CVC and MAP returned to pre-exercise levels in the PNB trial but remained reduced in the control trial. These findings suggest that central command, as well as muscle metabo-sensitive afferent stimulation, contributes to forearm cutaneous vascular responses in heat stressed humans. PMID- 15790666 TI - Angiotensin II regulation of ovine fetoplacental artery endothelial functions: interactions with nitric oxide. AB - During normal pregnancy, elevated angiotensin II (Ang II) concentrations in the maternal and fetal circulations are associated with dramatic increases in placental angiogenesis and blood flow. Much is known about a local renin angiotensin system within the uteroplacental vasculature. However, the roles of Ang II in regulating fetoplacental vascular functions are less well defined. In the fetal placenta, the overall in vivo vasoconstrictor responses of the blood vessels to Ang II infusion is thought to be less than that in its maternal counterpart, even though infused Ang II induces vasoconstriction. Recent data from our laboratories suggest that Ang II stimulates cell proliferation and increases endothelial nitric oxide synthase (eNOS) and production of nitric oxide (NO) in ovine fetoplacental artery endothelial cells. These data imply that elevations of the known vasoconstrictor Ang II in the fetal circulation may indeed play a role in the marked increases in fetoplacental angiogenesis and that Ang II-elevated endothelial NO production may partly attenuate Ang II-induced vasoconstriction on vascular smooth muscle. Together with both of these processes, the high levels of Ang II in the fetal circulation may serve to modulate overall fetoplacental vascular resistance. In this article, we review currently available data on the expression of Ang II receptors in the ovine fetal placenta with particular emphasis on the effects of Ang II on ovine fetoplacental endothelium. The potential cellular mechanisms underlying the regulation of Ang II on endothelial growth and vasodilator production are discussed. PMID- 15790667 TI - A1152D mutation of the Na+ channel causes paramyotonia congenita and emphasizes the role of DIII/S4-S5 linker in fast inactivation. AB - Missense mutations in the human skeletal muscle Na+ channel alpha subunit (hSkM1) are responsible for a number of muscle excitability disorders. Among them, paramyotonia congenita (PC) is characterized by episodes of muscle stiffness induced by cold and aggravated by exercise. We have identified a new PC associated mutation, which substitutes aspartic acid for a conserved alanine in the S4-S5 linker of domain III (A1152D). This residue is of particular interest since its homologue in the rat brain type II Na+ channel has been suggested as an essential receptor site for the fast inactivation particle. To identify the biophysical changes induced by the A1152D mutation, we stably expressed hSkM1 mutant or wild-type (WT) channels in HEK293 (human embryonic kidney) cells, and recorded whole-cell Na+ currents with the patch-clamp technique. Experiments were performed both at 21 and 11 degrees C to better understand the sensitivity to cold of paramyotonia. The A1152D mutation disrupted channel fast inactivation. In comparison to the WT, mutant channels inactivated with slower kinetics and displayed a 5 mV depolarizing shift in the voltage dependence of the steady state. The other noticeable defect of A1152D mutant channels was an accelerated rate of deactivation from the inactivated state. Decreasing temperature by 10 degrees C amplified the differences in channel gating kinetics between mutant and WT, and unveiled differences in both the sustained current and channel deactivation from the open state. Overall, cold-exacerbated mutant defects may result in a sufficient excess of Na+ influx to produce repetitive firing and myotonia. In the light of previous reports, our data point to functional as well as phenotypic differences between mutations of conserved S4-S5 residues in domains II and III of the human skeletal muscle Na+ channel. PMID- 15790669 TI - Haplotype analysis of the RAGE gene: identification of a haplotype marker for diabetic nephropathy in type 2 diabetes mellitus. AB - BACKGROUND: Diabetic nephropathy (DN) represents a devastating complication of diabetes. Family clustering, heterogeneity in the onset and progression and results of segregation studies indicate that susceptibility to DN is a complex trait. METHODS: Common single nucleotide polymorphisms in the RAGE (receptor of advanced glycation end-products) gene (-429T/C, -374T/A, G82S, 1704G/T, 2184A/G and 2245G/A) were studied in the association study comprising 605 Caucasian subjects by means of haplotype analysis in order to identify an eventual haplotype marker for DN in type 2 diabetes. Haplotypes were constructed computationally; frequencies were compared among groups of subjects with type 2 diabetes (DM) and DN, diabetics without DN and non-diabetics. Survival analysis was carried out to ascertain whether certain RAGE haplotypes influence onset of DN in type 2 diabetics. RESULTS: Significant differences in haplotype frequencies among DM + DN vs DM non-DN and non-DM groups were found (P = 0.0007 and 0.0013, respectively; permutation test). Frequency of the RAGE(2) haplotype containing minor alleles in positions -429 and 2184 (CTGGGG) in the DN group was significantly higher than in the two control groups (21.7% vs 12.8% and 13.8%, both P(corr)<0.003; two-tail Fisher exact test); odds ratios 1.65 [95% confidence interval (CI): 1.08-2.50; P = 0.020] and 1.79 (95% CI: 1.22-2.62; P = 0.003), respectively. In survival analysis, duration of diabetes until the onset of DN (e.g. appearance of persistent proteinuria) was significantly different among RAGE(2) diplotype groups (P<0.05); median DN-free interval was 9.6 years in RAGE(2) +/+ homozygotes, 15.2 years in +/- heterozygotes and 17.0 years in the -/ combination. CONCLUSIONS: The RAGE(2) haplotype is associated with DN in type 2 diabetics and with earlier DN onset and, thus, can be regarded a marker for DN. PMID- 15790668 TI - Development of cardiovascular function in the horse fetus. AB - In mammals, the mechanisms regulating an increase in fetal arterial blood pressure with advancing gestational age remain unidentified. In all species studied to date, the prepartum increase in fetal plasma cortisol has an important role in the maturation of physiological systems essential for neonatal survival. In the horse, the prepartum elevation in fetal cortisol and arterial blood pressure are delayed relative to other species. Hence, the mechanisms governing the ontogenic increase in arterial blood pressure in the horse fetus may mature much closer to term than in other fetal animals. In the chronically instrumented pony mare and fetus, this study investigated how changes in fetal peripheral vascular resistance, in plasma concentrations of noradrenaline, adrenaline and vasopressin, and in the maternal-to-fetal plasma concentration gradient of oxygen and glucose relate to the ontogenic changes in fetal arterial blood pressure and fetal plasma cortisol concentration as term approaches. The data show that, towards term in the horse fetus, the increase in arterial blood pressure occurs together with reductions in metatarsal vascular resistance, elevations in plasma concentrations of cortisol, vasopressin, adrenaline and noradrenaline, and falls in the fetal : maternal ratio of blood P(a,O(2)) and glucose concentration. Correlation analysis revealed that arterial blood pressure was positively related with plasma concentrations of vasopressin and noradrenaline, but not adrenaline in the fetus, and inversely related to the fetal : maternal ratio of blood P(a,O(2)), but not glucose, concentration. This suggests that increasing vasopressinergic and noradrenergic influences as well as changes in oxygen availability to the fetus and uteroplacental tissues may contribute to the ontogenic increase in fetal arterial blood pressure towards term in the horse. PMID- 15790670 TI - In vitro activity of tigecycline against ampicillin-resistant Haemophilus influenzae isolates. PMID- 15790671 TI - Oxygen accessibility and iron levels are critical factors for the antifungal action of ciclopirox against Candida albicans. AB - OBJECTIVES: Ciclopirox is a topical antifungal agent of the hydroxypyridone class whose mode of action is poorly understood. In order to elucidate the mechanism of action of ciclopirox, we analysed the growth, cellular integrity, biochemical properties, viability and transcriptional profile of the polymorphic yeast Candida albicans following exposure to this antifungal agent. METHODS: Multiple biochemical assays served to identify factors that were critical for antifungal activity and to identify proteins whose activities changed in drug-exposed cells. Genome-wide transcriptional profiling was used to identify genes that were up regulated in response to the cellular effects of the drug. RESULTS: Ciclopirox inhibited growth of C. albicans yeast and hyphal cells in a dose-dependent manner. This effect was reduced (i) by the addition of iron ions or the metabolic inhibitor 2-deoxy-D-glucose to growth media, (ii) in media that lacked glucose, and (iii) for cells that were pre-incubated with hydrogen peroxide or menadione [which caused induction of proteins involved in detoxification of reactive oxygen species (ROS)]. In contrast, cells pre-cultured under poor oxygen conditions (which had decreased activity of proteins involved in ROS detoxification) were more susceptible to ciclopirox. Treatment with ciclopirox did not directly cause cell membrane damage and did not change intracellular levels of ATP. Finally, the transcriptional profiling pattern of drug-treated cells strongly resembled iron limited conditions. CONCLUSIONS: These data indicate that metabolic activity, oxygen accessibility and iron levels are critical parameters in the mode of action of ciclopirox olamine. PMID- 15790672 TI - Anti-proliferative synergy of lysophospholipid analogues and ketoconazole against Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae): cellular and ultrastructural analysis. AB - OBJECTIVES: Investigation of the antiproliferative synergy of the lysophospholipid analogues (LPAs) edelfosine, ilmofosine and miltefosine with the ergosterol biosynthesis inhibitor ketoconazole against Trypanosoma cruzi. METHODS: The effect of LPAs, ketoconazole and their combination was evaluated against epimastigotes and intracellular amastigotes by the parameter IC50 leading to construction of isobolograms, for determination of a synergic effect. For epimastigotes, ultrastructural damage induced by these treatments was evaluated by transmission and scanning electron microscopy. RESULTS: Synergy was confirmed against both epimastigotes and amastigotes of the parasite. Edelfosine or ketoconazole alone induced morphological alterations in the plasma membrane and reservosomes of the parasites, while in combination, they also led to severe mitochondrial damage, formation of autophagic structures and multinucleation. Scanning electron microscopy confirmed the effect at the plasma membrane and also revealed alterations in the shape of the parasites. CONCLUSIONS: Our results describe the synergic anti-proliferative effect of LPAs and ketoconazole against epimastigotes and intracellular amastigotes and suggest that in epimastigotes, plasma membrane, reservosomes and mitochondria are targets of these drugs, possibly by interference with lipid metabolism. PMID- 15790673 TI - Time trends in the treatment and survival of recurrences from colorectal cancer. AB - BACKGROUND: Little is known about the management of recurrences from colorectal cancer at a population level. MATERIALS AND METHODS: Data was obtained from the population-based cancer registry of Cote d'Or (Burgundy, France) over a 28-year period. Univariate and multivariate analyses were performed to analyse trends in treatment and survival for local recurrence and distant metastases. RESULTS: The proportion of patients resected for cure increased from 6.7% (1976-1984) to 23.7% (1994-2003; P <0.001) for distant metastases and from 15.9% to 58.1% (P <0.001) for local recurrence. Age and period of diagnosis were independent factors associated with a resection for cure. Rectal cancer local recurrence was less often resected for cure than colon cancer local recurrence (P=0.05). Long-term survival was observed only after resection for cure: 5-year relative survival rates were 36.1% for local recurrence and 24.0% for distant metastases. In the multivariate analysis, survival decreased with age and increased over time but significantly only over the last study period. Surgical resection and palliative chemotherapy were other determinants of prognosis for distant metastases whereas surgical resection and palliative radiotherapy did influence the prognosis for local recurrence. CONCLUSION: Substantial advances in the management of recurrences have been achieved over time. More effective treatments and mass screening represent promising approaches to decrease this problem. PMID- 15790674 TI - Volumetric preload measurement by thermodilution: a comparison with transoesophageal echocardiography. AB - BACKGROUND: End-diastolic volume indices determined by transpulmonary thermodilution and pulmonary artery thermodilution may give a better estimate of left ventricular preload than pulmonary capillary wedge pressure monitoring. The aim of this study was to compare volume preload monitoring using the two different thermodilution techniques with left ventricular preload assessment by transoesophageal echocardiography (TOE). METHODS: Twenty patients undergoing elective cardiac surgery with preserved left-right ventricular function were studied after induction of anaesthesia. Conventional haemodynamic variables, global end-diastolic volume index using the pulse contour cardiac output (PiCCO) system (GEDVI(PiCCO)), continuous end-diastolic volume index (CEDVI(PAC)) measured by a modified pulmonary artery catheter (PAC), left ventricular end diastolic area index (LVEDAI) using TOE and stroke volume indices (SVI) were recorded before and 20 and 40 min after fluid replacement therapy. Analysis of variance (Bonferroni-Dunn), Bland-Altman analysis and linear regression were performed. RESULTS: GEDVI(PiCCO), CEDVI(PAC), LVEDAI and SVI(PiCCO/PAC) increased significantly after fluid load (P<0.05). An increase >10% for GEDVI(PiCCO) and LVEDAI was observed in 85% and 90% of the patients compared with 45% for CEDVI(PAC). Mean bias (2 SD) between percentage changes (delta) in GEDVI(PiCCO) and deltaLVEDAI was -3.2 (17.6)% and between deltaCEDVI(PAC) and deltaLVEDAI -8.7 (30.0)%. The correlation coefficient (r2) for deltaGEDVI(PiCCO) vs deltaLVEDAI was 0.658 and for deltaCEDVI(PAC) vs deltaLVEDAI 0.161. The relationship between deltaGEDVI(PiCCO) and deltaSVI(PiCCO) was stronger (r2=0.576) than that between deltaCEDVI(PAC) and deltaSVI(PAC) (r2=0.267). CONCLUSION: GEDVI assessed by the PiCCO system gives a better reflection of echocardiographic changes in left ventricular preload, in response to fluid replacement therapy, than CEDVI measured by a modified PAC. PMID- 15790675 TI - Onset of acetaminophen analgesia: comparison of oral and intravenous routes after third molar surgery. AB - BACKGROUND: The purpose of this randomized double-blind study was to compare the efficacy and safety of propacetamol 2 g (an i.v. acetaminophen 1 g formulation) administered as a 2-min bolus injection (n=50) or a 15-min infusion (n=50) with oral acetaminophen 1 g (n=50) or placebo (n=25) for analgesia after third molar surgery in patients with moderate to severe pain after impacted third molar removal. METHODS: All patients were evaluated for efficacy during the initial 6 h period after treatment administration (T(0)) and for safety during the entire week after T(0). RESULTS: The onset of analgesia after propacetamol was shorter (3 min for bolus administration, 5 min for 15-min infusion) than after oral acetaminophen (11 min). Active treatments were significantly better for all parameters (pain relief, pain intensity, patient's global evaluation, duration of analgesia) than placebo (P<0.05). Adverse events were more frequent after propacetamol, especially pain at the injection site. Propacetamol bolus resulted in a much higher incidence of local adverse events than the infusion (propacetamol bolus 90% vs propacetamol infusion 52%) with no clinically significant benefits in terms of analgesic efficacy. CONCLUSION: I.V. propacetamol, administered as a 15-min infusion, is a fast-acting analgesic agent. It is more effective in terms of onset of analgesia than a similar dose of oral acetaminophen. PMID- 15790676 TI - Plasticity of face processing in infancy. AB - Experience plays a crucial role for the normal development of many perceptual and cognitive functions, such as speech perception. For example, between 6 and 10 months of age, the infant's ability to discriminate among native speech sounds improves, whereas the ability to discriminate among foreign speech sounds declines. However, a recent investigation suggests that some experience with non native languages from 9 months of age facilitates the maintenance of this ability at 12 months. Nelson has suggested that the systems underlying face processing may be similarly sculpted by experience with different kinds of faces. In the current investigation, we demonstrate that, in human infants between 6 and 9 months of age, exposure to non-native faces, in this case, faces of Barbary macaques (Macaca sylvanus), facilitates the discrimination of monkey faces, an ability that is otherwise lost around 9 months of age. These data support, and further elucidate, the role of early experience in the development of face processing. PMID- 15790677 TI - Floral asymmetry involves an interplay between TCP and MYB transcription factors in Antirrhinum. AB - To understand how genes control floral asymmetry, we have isolated and analyzed the role of the RADIALIS (RAD) gene in Antirrhinum. We show that the RAD gene encodes a small MYB-like protein that is specifically expressed in the dorsal region of developing flowers. RAD has a single MYB-like domain that is closely related to one of the two MYB-like domains of DIV, a protein that has an antagonistic effect to RAD on floral development. Interactions between RAD and other genes indicate that floral asymmetry depends on the interplay between two pairs of transcription factors. First, a pair of TCP proteins is expressed in dorsal regions of the floral meristem, leading to the activation of RAD in the dorsal domain. The RAD MYB-like protein then antagonizes the related DIV MYB-like protein, preventing DIV activity in dorsal regions. In addition to its role in dorsal regions, RAD acts nonautonomously on lateral regions either directly, through RAD protein movement, or indirectly, through a signaling molecule. PMID- 15790678 TI - Cyclin-dependent kinase 6 associates with the androgen receptor and enhances its transcriptional activity in prostate cancer cells. AB - Cyclin-dependent kinase 6 (CDK6) binds to and is activated by cyclin D1 and thereby enhances the transition of cells through the G1 phase of the cell cycle. The present study indicates that, in human prostate cancer cells, CDK6 can also bind to the androgen receptor (AR) and stimulate its transcriptional activity in the presence of dihydrotestosterone. This effect of CDK6 does not require its kinase activity and is inhibited by cyclin D1 and p16INK4a. The T877A mutant of the AR frequently found in advanced cases of prostate cancer displays an exaggerated stimulation of transcriptional activity by CDK6. Androgen-sensitive LNCaP prostate cancer cells engineered to stably overexpress CDK6 display increased expression of the prostate-specific antigen and enhanced growth in the presence of dihydrotestosterone. CDK6 is present in the chromatin structure of these cells in association with the AR and the promoter region of the prostate specific antigen gene. These findings suggest that CDK6 may play an important role in the development and/or progression of a subset of human prostate cancers by stimulating the activity of the AR. PMID- 15790679 TI - Adult human hematopoietic stem cells produce neurons efficiently in the regenerating chicken embryo spinal cord. AB - Hematopoietic stem cells (HSCs) have been proposed as a potential source of neural cells for use in repairing brain lesions, but previous studies indicate a low rate of neuronal differentiation and have not provided definite evidence of neuronal phenotype. To test the neurogenic potential of human HSCs, we implanted CD34+ HSCs from adult human bone marrow into lesions of the developing spinal cord in the chicken embryo and followed their differentiation by using immunohistochemistry, retrograde labeling, and electrophysiology. We find that human cells derived from the implanted population express the neuronal markers NeuN and MAP2 at substantially higher rates than previously reported. We also find that these cells exhibit neuronal cytoarchitecture, extend axons into the ventral roots or several segments in length within the spinal white matter, are decorated with synaptotagmin+ and GABA+ synaptic terminals, and exhibit active membrane properties and spontaneous synaptic potentials characteristic of functionally integrated neurons. Neuronal differentiation is accompanied by loss of CD34 expression. Careful examination with confocal microscopy reveals no signs of heterokaryons, and human cells never express a chicken-specific antigen, suggesting that fusion with host chicken cells is unlikely. We conclude that the microenvironment in the regenerating spinal cord of the chicken embryo stimulates substantial proportions of adult human HSCs to differentiate into full-fledged neurons. This may open new possibilities for a high-yield production of neurons from a patient's own bone marrow. PMID- 15790680 TI - A long-range attraction between aggregating 3T3 cells mediated by near-infrared light scattering. AB - At what range can a mammalian cell sense the presence of another cell and through what medium? To approach these questions, the formation of aggregates of a 3T3 cell variant (3T3x cells) grown on solid substrates was studied. Each of the aggregates consisted of cells that, at the time of their seeding, were single and located randomly. Yet somehow they seemed to detect each other within a certain range (R(a)) and move together to form aggregates. The article describes a simple assay to measure the value of R(a). When applied to 3T3x cells with altered intensities of near-infrared light scattering (I(sc)) the assay showed that (i) R(a) was much larger than one cell diameter, and (ii) R(a) was directly related to I(sc). The results suggest that near-infrared light scattering by the cells mediate a long-range attraction between them, which does not require physical contact and enables them to detect each other's presence. PMID- 15790681 TI - Foxp3 interacts with nuclear factor of activated T cells and NF-kappa B to repress cytokine gene expression and effector functions of T helper cells. AB - Scurfy mice, which are deficient in a functional Foxp3, exhibit a severe lymphoproliferative disorder and display generalized over-production of cytokines. Here, we show that, among the Foxp transcriptional factor family, which includes Foxp1, Foxp2, and Foxp3, only Foxp3 has the ability to inhibit IL 2, IL-4, and IFN-gamma production by primary T helper cells. We found that Foxp3 physically associates with the Rel family transcription factors, nuclear factor of activated T cells (NFAT) and NF-kappaB, and blocks their ability to induce the endogenous expression of their target genes, including key cytokine genes. More importantly, T cells derived from scurfy mice have a dramatic increase in nuclear factor of activated T cells (NFAT) and NF-kappa B transcriptional activity compared with the T cells derived from WT mice. Furthermore, complementation of Foxp3 in scurfy-derived T cells lowers the NFAT and NF-kappa B transcriptional activity to the physiological level. Finally, we show that myelin proteolipid protein-specific autoreactive T cells transduced with Foxp3 cannot mediate experimental autoimmune encephalomyelitis, providing further support that Foxp3 suppresses the effector function of autoreactive T cells. Foxp3 has already been associated with the generation of CD4(+)CD25+ regulatory T cells; our data additionally demonstrate that Foxp3 suppresses the effector functions of T helper cells by directly inhibiting the activity of two key transcription factors, NFAT and NF-kappa B, which are essential for cytokine gene expression and T cell functions. PMID- 15790683 TI - Effects of topical anesthesia on upper airway resistance during wake-sleep transitions. AB - Deformation of the upper airway (UA) by negative transmural pressure alters the activity of UA mechanoreceptors, causing a reflex increase in UA muscle activity. Topical anesthesia of the UA mucosa, which greatly reduces this reflex response, causes an increase in UA resistance during stage 2 sleep. We hypothesized that topical anesthesia of the UA mucosa would predispose to UA instability at sleep onset and, therefore, examined the effect of UA anesthesia on pharyngeal resistance (Rph) in stage 1 sleep. Eleven normal, healthy volunteers were instrumented to record standard polysomnographic variables, respiratory airflow, and UA pressure at the nasal choanae and the epiglottis. Subjects were permitted to sleep until stable stage 2 sleep was reached and were then awoken. This procedure was repeated three times to obtain reproducible wake-sleep transitions. The UA mucosa was then anesthetized with 10% lidocaine to the oropharynx and laryngopharynx, and the pharyngeal mechanics were studied during the subsequent wake-sleep transition. Three subjects were excluded because of failure to resume sleep postanesthesia. Rph was significantly higher after anesthesia during stage 1 sleep [2.88 +/- 0.77 cmH(2)O.l(-1).s (mean +/- SE)] compared with control (0.95 +/- 0.35 cmH(2)O.l(-1).s; P < 0.05), but there was no difference during wakefulness. Furthermore, there was a significant rise in Rph at wake-to-sleep transitions and a significant fall in Rph at sleep-to-wake transitions after anesthesia (P < 0.05) but not in the control condition. We conclude that sensory receptors in the UA mucosa contribute to the maintenance of UA patency at wake sleep transition in normal humans. PMID- 15790684 TI - Quantification of muscle fiber strain during in vivo repetitive stretch shortening cycles. AB - Muscles subjected to lengthening contractions exhibit evidence of subcellular disruption, arguably a result of fiber strain magnitude. Due to the difficulty associated with measuring fiber strains during lengthening contractions, fiber length estimates have been used to formulate relationships between the magnitude of injury and mechanical measures such as fiber strain. In such protocols, the series compliance is typically minimized by removing the distal tendon and/or preactivating the muscle. These in vitro and in situ experiments do not represent physiological contractions well where fiber strain and muscle strain may be disassociated; thus the mechanisms of in vivo muscle injury remain elusive. The purpose of this paper was to quantify fiber strains during lengthening contractions in vivo and assess the potential role of fiber strain in muscle injury following repetitive stretch-shortening cycles. Using intact New Zealand White rabbit dorsiflexors, fiber strain and joint torque were measured during 50 stretch-shortening cycles. We were able to show that fiber length changes are disassociated from muscle tendon unit length changes and that complex fiber dynamics during these cycles prevent easy estimates of fiber strains. In addition, fiber strains vary, depending on how they are defined, and vary from repetition to repetition, thereby further complicating the potential relationship between muscle injury and fiber strain. We conclude from this study that, during in vivo stretch-shortening cycles, the relationship between fiber strain and muscle injury is complex. This is due, in part, to temporal effects of repeated loading on fiber strain magnitude that may be explained by an increasing compliance of the contractile element as exercise progresses. PMID- 15790682 TI - Dramatic variation of the vomeronasal pheromone receptor gene repertoire among five orders of placental and marsupial mammals. AB - Pheromones are chemicals emitted and sensed by conspecifics to elicit social and sexual responses and are perceived in terrestrial vertebrates primarily by the vomeronasal organ (VNO). Pheromone receptors in the mammalian VNO are encoded by the V1R and V2R gene superfamilies. The V1R superfamily contains 187 and 102 putatively functional genes in the mouse and rat, respectively. To investigate whether this large repertoire size is typical among mammals with functional VNOs, we here describe the V1R repertoires of dog, cow, and opossum based on their draft genome sequences. The dog and cow have only 8 and 32 intact V1R genes, respectively. Thus, the intact V1R repertoire size varies by at least 23-fold among placental mammals with functional VNOs. To our knowledge, this size ratio represents the greatest among-species variation in gene family size of all mammalian gene families. Phylogenetic analysis of placental V1R genes suggests multiple losses of ancestral genes in carnivores and artiodactyls and gains of many new genes by gene duplication in rodents, manifesting massive gene births and deaths. We also identify 49 intact opossum V1R genes and discover independent expansions of the repertoire in placentals and marsupials. We further show a concordance between the V1R repertoire size and the complexity of VNO morphology, suggesting that the latter could indicate the sophistication of pheromone communications within species. In sum, our results demonstrate tremendous diversity and rapid evolution of mammalian V1R gene inventories and caution the generalization of VNO biology from rodents to all mammals. PMID- 15790685 TI - In vivo insulin signaling through PI3-kinase is impaired in skeletal muscle of adult rat offspring exposed to ethanol in utero. AB - It is now known that prenatal ethanol (EtOH) exposure is associated with impaired glucose tolerance and insulin resistance in rat offspring, but the underlying mechanism(s) is not known. To test the hypothesis that in vivo insulin signaling through phosphatidylinositol 3 (PI3)-kinase is reduced in skeletal muscle of adult rat offspring exposed to EtOH in utero, we gave insulin intravenously to these rats and probed steps in the PI3-kinase insulin signaling pathway. After insulin treatment, EtOH-exposed rats had decreased tyrosine phosphorylation of the insulin receptor beta-subunit and of insulin receptor substrate-1 (IRS-1), as well as reduced IRS-1-associated PI3-kinase in the gastrocnemius muscle compared with control rats. There was no significant difference in basal or insulin stimulated Akt activity between EtOH-exposed rats and controls. Insulin stimulated PKC isoform zeta phosphorylation and membrane association were reduced in EtOH-exposed rats compared with controls. Muscle insulin binding and peptide contents of insulin receptor, IRS-1, p85 subunit of PI3-kinase, Akt/PKB, and atypical PKC isoform zeta were not different between EtOH-exposed rats and controls. Thus insulin resistance in rat offspring exposed to EtOH in utero may be explained, at least in part, by impaired insulin signaling through the PI3 kinase pathway in skeletal muscle. PMID- 15790686 TI - Dual effect of deafferentation on contractile characteristics and sarcoplasmic reticulum properties in rat soleus fibers. AB - The neural message is known to play a key role in muscle development and function. We analyzed the specific role of the afferent message on the functional regulation of two subcellular muscle components involved in the contractile mechanism: the contractile proteins and the sarcoplasmic reticulum (SR). Rats were submitted to bilateral deafferentation (DEAF group) by section of the dorsal roots L(3) to L(5) after laminectomy. Experiments were carried out in single skinned fibers of the soleus muscle. The maximal force developed by the contractile proteins was increased in the DEAF group compared with control, despite a decrease in muscle mass by 17%. The tension-pCa relationship was shifted toward lower calcium (Ca(2+)) concentrations. Different functional properties of the SR of DEAF soleus were examined by using caffeine-induced contractions. The caffeine sensitivity of the Ca(2+) release was decreased after deafferentation and ryanodine receptor 1 isoform was expressed at a lower level. The rate of Ca(2+) uptake was only slightly increased. The results underlined the dual effect of the afferent input on the functional regulation of both contractile proteins and SR. PMID- 15790687 TI - Optical monitoring of bubble size and shape in a pulsating bubble surfactometer. AB - The pulsating bubble surfactometer (PBS) is often used for in vitro characterization of exogenous lung surfactant replacements and lung surfactant components. However, the commercially available PBS is not able to dynamically track bubble size and shape. The PBS therefore does not account for bubble growth or elliptical bubble shape that frequently occur during device use. More importantly, the oscillatory volume changes of the pulsating bubble are different than those assumed by the software of the commercial unit. This leads to errors in both surface area and surface tension measurements. We have modified a commercial PBS through the addition of an image-acquisition system, allowing real time determination of bubble size and shape and hence the accurate tracking of surface area and surface tension. Compression-expansion loops obtained with the commercially available PBS software were compared with those provided by the image-analysis system for dipalmitoylphosphatidylcholine, Infasurf, and Tanaka lipids (dipalmitoylphosphatidylcholine-palmitoyloleoylphosphatidyl-glycerol palmitic acid, 68:22:9) at concentrations of 0.1 and 1.0 mg/ml and at frequencies of 1 and 20 cycles/min. Whereas minimum surface tension as determined by the image-analysis system is similar to that measured by the commercially available software, the maximum surface tension and the shapes of the interfacial area surface tension loops are quite different. Differences are attributable to bubble drift, nonsinusoidal volume changes, and variable volume excursions seen with the modified system but neglected by the original system. Image analysis reveals that the extent of loop hysteresis is greatly overestimated by the commercial device and that an apparent, rapid increase in surface tension upon film expansion seen in PBS loops is not observed with the image-analysis system. The modified PBS system reveals new dynamic characteristics of lung surfactant preparations that have not previously been reported. PMID- 15790689 TI - Elastic properties of human Achilles tendon are correlated to muscle strength. AB - The purpose of this study was to investigate whether the mechanical properties of the Achilles tendon were correlated to muscle strength in the triceps surae in humans. Twenty-four men and twelve women exerted maximal voluntary isometric plantar flexion (MVIP) torque. The elongation (DeltaX) and strain of the Achilles tendon (epsilon), the proximal part of which is the composite of the gastrocnemius tendon and the soleus aponeurosis, at MVIP were determined from the displacement of the distal myotendinous junction of the medial gastrocnemius using ultrasonography. The Achilles tendon force at MVIP (F) was calculated from the MVIP torque and the Achilles tendon moment arm. There were no significant differences in either the F-DeltaX or F-epsilon relationships between men and women. DeltaX and epsilon were 9.8 +/- 2.6 mm and 5.3 +/- 1.6%, respectively, and were positively correlated to F (r = 0.39, P < 0.05; r = 0.39, P < 0.05), which meant that subjects with greater muscle strength could store more elastic energy in the tendon. The regression y-intercepts for the F-DeltaX (P < 0.01) and F epsilon (P < 0.05) relationship were significantly positive. These results might indicate that the Achilles tendon was stiffer in subjects with greater muscle strength, which may play a role in reducing the probability of tendon strain injuries. It was suggested that the Achilles tendon of subjects with greater muscle strength did not impair the potential for storing elastic energy in tendons and may be able to deliver the greater force supplied from a stronger muscle more efficiently. Furthermore, the difference in the Achilles tendon mechanical properties between men and women seemed to be correlated to the difference in muscle strength rather than gender. PMID- 15790688 TI - Bubble splitting in bifurcating tubes: a model study of cardiovascular gas emboli transport. AB - The transport of long gas bubbles, suspended in liquid, through symmetric bifurcations, is investigated experimentally and theoretically as a model of cardiovascular gas bubble transport in air embolism and gas embolotherapy. The relevant dimensionless parameters in the models match the corresponding values for arteries and arterioles. The effects of roll angle (the angle the plane of the bifurcation makes with the horizontal), capillary number (a dimensionless indicator of flow), and bubble volume (or length) on the splitting of bubbles as they pass through the bifurcation are examined. Splitting is observed to be more homogenous at higher capillary numbers and lower roll angles. It is shown that, at nonzero roll angles, there is a critical value of the capillary number below which the bubbles do not split and are transported entirely into the upper branch. The value of the critical capillary number increases with roll angle and parent tube diameter. A unique bubble motion is observed at the critical capillary number and for slightly slower flows: the bubble begins to split, the meniscus in the lower branch then moves backward, and finally the entire bubble enters the upper branch. These findings suggest that, in large vessels, emboli tend to be transported upward unless flow is unusually strong but that a more homogeneous distribution of emboli occurs in smaller vessels. This corresponds to previous observations that air emboli tend to lodge in the upper regions of the lungs and suggests that relatively uniform infarction of tumors by gas embolotherapy may be possible. PMID- 15790690 TI - Effects of tidal volume and respiratory frequency on lung lymph flow. AB - Ventilation (V) increases lung lymph flow (Ql), but the separate effects of tidal volume (Vt) and frequency (f) and the role of V-induced changes in edema formation are poorly understood. An isolated, in situ sheep lung preparation was used to examine these effects. In eight sheep with f = 10 min(-1), results obtained during 30-min periods with Vt = 5 or 20 ml/kg were compared with values obtained during bracketed 30-min control periods (Vt = 12.5 ml/kg). Eight other sheep with constant Vt (12.5 ml/kg) were studied at f = 5 or 20 min(-1) and compared with f = 10 min(-1). Three additional groups of six sheep were perfused for 100 min with control V (10 ml/kg, 10 min(-1)). Vt was then kept constant or changed to 20 or 3 ml/kg during a second 100-min period. Increases in Vt or f increased Ql and vice versa, without corresponding effects on the rate of edema formation. For the same change in V, changing Vt had a greater effect on Ql than changing f. The change in Ql caused by an increase in Vt was significantly greater after the accumulation of interstitial edema. The change in Ql caused by a sustained increase in Vt was transient and did not correlate with the rate of edema formation, suggesting that V altered Ql through direct mechanical effects on edema-filled compartments and lymphatic vessels rather than through V-induced changes in fluid filtration. PMID- 15790691 TI - Autonomic dysreflexia during sperm retrieval in spinal cord injury: influence of lesion level and sildenafil citrate. AB - Autonomic dysreflexia (AD) can occur during penile vibratory stimulation in men with spinal cord injury, but this is variable, and the association with lesion level is unclear. The purpose of this study was to characterize the cardiovascular responses to penile vibratory stimulation in men with spinal cord injury. We hypothesized that those with cervical injuries would demonstrate a greater degree of AD compared with men with thoracic injuries. We also questioned whether the rise in blood pressure could be attenuated by sildenafil citrate. Participants were classified as having cervical (n = 8) or thoracic (n = 5) injuries. While in a supine position, subjects were instrumented with an ECG, and arterial blood pressure was determined beat by beat. Subjects reported to the laboratory twice and received an oral dose of sildenafil citrate (25-100 mg) or no medication. Penile vibratory stimulation was performed using a handheld vibrator to the point of ejaculation. At ejaculation during the nonmedicated trials, the cervical group had a significant decrease in heart rate (-5-10 beats/min) and increase in mean arterial blood pressure (+70-90 mmHg) relative to resting conditions, whereas the thoracic group had significant increases in both heart rate (+8-15 beats/min) and mean arterial pressure (+25-30 mmHg). Sildenafil citrate had no effect on the change in heart rate or mean arterial pressure in either group. In summary, men with cervical injuries had more pronounced AD during penile vibratory stimulation than men with thoracic injuries. Administration of sildenafil citrate had no effect on heart rate or blood pressure during penile vibratory stimulation in men with spinal cord injury. PMID- 15790692 TI - Neonatal maternal separation enhances phrenic responses to hypoxia and carotid sinus nerve stimulation in the adult anesthetized rat. AB - In awake animals, our laboratory recently showed that the hypoxic ventilatory response of adult male (but not female) rats previously subjected to neonatal maternal separation (NMS) is 25% greater than controls (Genest SE, Gulemetova R, Laforest S, Drolet G, and Kinkead R. J Physiol 554: 543-557, 2004). To begin mechanistic investigations of the effects of this neonatal stress on respiratory control development, we tested the hypothesis that, in male rats, NMS enhances central integration of carotid body chemoafferent signals. Experiments were performed on two groups of adult male rats. Pups subjected to NMS were placed in a temperature-controlled incubator 3 h/day from postnatal day 3 to postnatal day 12. Control pups were undisturbed. At adulthood (8-10 wk), rats were anesthetized (urethane; 1.6 g/kg), paralyzed, and ventilated with a hyperoxic gas mixture [inspired O2 fraction (Fi(O2)) = 0.5], and phrenic nerve activity was recorded. The first series of experiments aimed to demonstrate that NMS-related enhancement of the inspiratory motor output (phrenic) response to hypoxia occurs in anesthetized animals also. In this series, rats were exposed to moderate, followed by severe, isocapnic hypoxia (Fi(O2) = 0.12 and 0.08, respectively, 5 min each). NMS enhanced both the frequency and amplitude components of the phrenic response to hypoxia relative to controls, thereby validating the use of this approach. In a second series of experiments, NMS increased the amplitude (but not the frequency) response to unilateral carotid sinus nerve stimulation (stimulation frequency range: 0.5-33 Hz). We conclude that enhancement of central integration of carotid body afferent signal contributes to the larger hypoxic ventilatory response observed in NMS rats. PMID- 15790693 TI - Legislative changes. PMID- 15790694 TI - Best practice no 183. Examination of parathyroid gland specimens. AB - The pathological examination of parathyroid glands is an essential component of the evaluation of hyperparathyroidism. Traditionally, this has involved intraoperative frozen sections during bilateral surgical exploration of the neck, to confirm removal of parathyroid tissue. With recent developments in imaging, some diseased glands can be localised preoperatively, enabling removal by minimally invasive, targetted surgery, with or without additional non histological intraoperative procedures to confirm the removal of all hyperfunctioning parathyroid tissue. This article reviews these developments and describes the ideal approach to reporting parathyroid specimens. PMID- 15790696 TI - Bacteraemia prediction in emergency medical admissions: role of C reactive protein. AB - AIM: To define the contribution made by C reactive protein (CRP) measurement to bacteraemia prediction in adults with medical emergencies in the UK. METHODS: This two year cohort study involved 6234 patients admitted as emergency cases to the acute medical or infectious diseases services of the Oxford Radcliffe Hospitals, in whom blood cultures were taken on arrival. The main outcome measures were bacteraemia risk associated with admission CRP concentrations, lymphocyte counts, and neutrophil counts. RESULTS: The quantitative associations between CRP concentration, admission lymphocyte count, and neutrophil count were defined. Risk of bacteraemia rose continuously as the CRP increased: no "cutoff" value was evident. Models examining combinations of CRP, neutrophil count, and lymphocyte count were developed and validated using a split sample technique. CRP contributed to a model including lymphocyte and neutrophil counts, but its effect was small. CRP alone performed no better than either a model combining lymphopenia and neutrophilia, or than lymphopenia alone. CONCLUSIONS: In patients with acute medical emergencies who are suspected of bacteraemia clinically, CRP concentrations, although associated with bacteraemia, have a limited role in bacteraemia prediction. PMID- 15790697 TI - Coxsackievirus B3 sequences in the myocardium of fatal cases in a cluster of acute myocarditis in Greece. AB - AIM: The investigation of three fatal cases during a nationwide cluster of cases of an upper respiratory tract infection (URTI) associated with myocarditis and/or pericarditis in Greece in 2002. METHODS: In the three women who died, necropsies were performed and tissue sections were taken for histological examination, antigen detection by immunohistochemistry and indirect immunofluorescence assay (IFA), amplification of viral genomes by nested reverse transcription polymerase chain reaction (RT-PCR), and sequence analysis. RESULTS: All samples showed histological signs of active myocarditis. Immunohistochemistry revealed the presence of the enterovirus VP1 family of proteins and IFA revealed the presence of coxsackievirus B3 antigen. Nested RT-PCR amplified enteroviral alleles of the 5'-untranslated region which were identical to each other and to the coxsackievirus B3 sequences. CONCLUSIONS: This study provides pathological evidence of enteroviral infection among fatal myocarditis cases in a nationwide URTI cluster of cases associated with myocarditis and/or pericarditis. PMID- 15790695 TI - Expression of cell adhesion molecules in oesophageal carcinoma and its prognostic value. AB - Oesophageal carcinoma remains a disease of poor prognosis. Surgical cure rates are compromised by the fact that most patients are diagnosed at a late stage of disease because of the delayed onset of symptoms, by which time metastases and organ infiltration may have already occurred. Thus, invasion and metastases play a key role in influencing patient survival, and the search for novel treatments may therefore hinge on gaining insight into the mechanisms controlling these processes. It has been established that the initial step in the metastatic cascade is the detachment of tumour cells from the primary tumour via dysregulation of normal cell-cell and cell-matrix interactions. Distinct proteins known as cell adhesion molecules (CAMs) mediate these interactions. In recent years, a plethora of information has contributed to the in depth understanding of these molecules. This review provides a brief description of five families of CAMs (cadherins, integrins, CD44, immunoglobulin superfamily, and selectins) and highlights their altered expression in relation both to prognosis and tumour behaviour in squamous cell carcinoma and adenocarcinoma of the oesophagus. PMID- 15790698 TI - Significant expression of IGFBP2 in breast cancer compared with benign lesions. AB - BACKGROUND/AIM: Insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs) play a role in the normal development of breast tissue, and possibly in breast cancer aetiology. IGFBP2, one of six members of the IGFBP superfamily, acts as regulator of the IGFs and has pleiotropic effects in normal and neoplastic tissues. Because IGFs have mitogenic effects on mammary epithelia, this study investigated IGFBP2 expression in mammary tissues of different benign and malignant entities. METHODS: Immunohistochemistry was used to study correlations between the presence and intensity of IGFBP2 staining and tumour type and grade, in addition to steroid hormone receptor status, in 120 breast specimens. Expression was measured by quantitative colour video image analysis and semiquantitative evaluation, and the measurements correlated well (r = 0.92; p<0.05). RESULTS: Both methods found no significant expression of IGFBP2 in normal glandular cells and hyperplasia (group I). Atypical hyperplasia showed a slightly increased cytoplasmic expression of IGFBP2, and carcinoma in situ showed a distinctive, membrane associated and cytoplasmic expression (group II). Infiltrating carcinomas strongly expressed cytoplasmic IGFBP2 (group III). There were significant differences between group I and II, and between group II and III. There were no significant differences between invasive lobular and invasive ductal carcinoma, or between grades I, II, and III within these entities. There was no significant correlation between IGFBP2 immunostaining and oestrogen or progesterone receptor positivity within the malignant group. CONCLUSIONS: IGFBP2 mitogenic signals of autocrine/paracrine regulatory mechanisms may be responsible for the growth of breast carcinomas and IGFBP2 may be an independent indicator of malignancy. PMID- 15790699 TI - Status of the p53, p16, RB1, and HER-2 genes and chromosomes 3, 7, 9, and 17 in advanced bladder cancer: correlation with adjacent mucosa and pathological parameters. AB - AIMS: To evaluate a panel of well known genetic alterations for frequency of changes in bladder cancer that could be considered genomic instability determinants or adjunctive prognostic predictors. METHODS: Fluorescence in situ hybridisation analysis was performed to evaluate chromosomes 3, 7, 9, and 17 and the 9p21 (p16), 17p13.1 (p53), 13q14 (RB1), and 17q11.2 (HER-2) chromosomal loci in 48 muscle invasive bladder cancer specimens and the adjacent normal mucosa. RESULTS: There were significant differences between the frequency of chromosome 7 monosomy/polysomy and 17 monosomy in the two groups (tumours and adjacent mucosa) (p = 0.004, p = 0.037, and p = 0.015, respectively). There were no differences in the frequency of gene deletions between tumours and the adjacent mucosa. 17q11.2 amplification was found in 14.5% of tumours examined, but not in the non malignant epithelium. Chromosome 3, 7, and 17 monosomy and the RB1 heterozygous deletion were significantly associated with stage T3-4 (p = 0.03, p = 0.04, p = 0.04, and p = 0.03, respectively). CONCLUSIONS: These results demonstrate the importance of chromosomes 3, 7, and 17 and gene alterations in bladder cancer progression, highlighting their usefulness as prognostic markers. Larger studies with longterm follow up of these patients are needed to determine the validity and clinical relevance of these genetic findings, and molecular prognostic markers should be incorporated into phase II and III trials to define their roles in predicting clinical outcome. PMID- 15790700 TI - Detection of BRAF mutations in colorectal tumours and peritoneal washings using a mismatch ligation assay. AB - AIMS: To detect cells bearing BRAF mutations in colorectal tumour samples and peritoneal washings, using a mismatch ligation assay (MLA). METHODS: DNA from 46 colorectal tumours was studied. Part of exon 15 of the BRAF gene was amplified using the polymerase chain reaction, and T-->A mutations at codon 600 were detected using MLA. When a mutation was detected, the same mutation was sought in peritoneal washings from that patient. RESULTS: BRAF mutations were detected in five of the 45 analysable tumour samples. In four cases, this result was confirmed by sequencing analysis. More tumours with BRAF mutations were Dukes' stage C or D rather than A or B (p < 0.02). Dilution experiments revealed that one mutant cell could be detected in 1000 normal cells. Cells with the same BRAF mutation were present in the peritoneal washing taken at the start of the operation in four of the five patients. CONCLUSIONS: MLA is a suitable technique for the detection of BRAF mutations, and allows the detection of small numbers of isolated tumour cells shed from the primary tumour. PMID- 15790701 TI - Inflammation and cytokeratin 7/20 staining of cardiac mucosa in young patients with and without Helicobacter pylori infection. AB - BACKGROUND: Both Helicobacter pylori and gastro-oesophageal reflux disease (GORD) may cause inflammation in cardiac mucosa. Intestinal metaplasia (IM) is found more often in GORD associated inflammation than in inflammation caused by H pylori, especially in young individuals. AIM: To examine morphological differences in chronic inflammation in these two conditions by immunohistochemistry. PATIENTS/METHODS: Tissue blocks from cardiac mucosa of patients <45 years were available as follows: 10 patients with chronic inflammation of cardiac mucosa (carditis) and H pylori gastritis (group 1); 10 patients with (possibly GORD related) carditis, but normal antrum and corpus (group 2); and 10 patients with non-inflamed cardiac mucosa and normal antrum and corpus (group 3). Haematoxylin and eosin staining and immunohistochemical staining for various inflammatory cells were performed for patients in groups 1 and 2 as follows: CD20 (B cells), CD3 (T cells), CD4 (T helper cells), CD8 (T suppressor cells), CD163 (macrophages), CD138 (plasma cells), and CD117 (mast cells). For all patients, cytokeratin 7/20 (CK7/20) staining was performed. RESULTS: No clear differences were seen in the morphology of chronic inflammation between groups 1 and 2. In both, plasma cells were most abundant. CK7/20 staining showed no differences between these groups. CONCLUSION: Helicobacter pylori negative (possibly GORD associated) and H pylori related carditis cannot be distinguished on a morphological basis. The stronger tendency towards IM in the first entity cannot be explained by differences in the type of inflammation. Barrett-type CK7/20 staining seems typical for cardiac mucosa, irrespective of the type of inflammation or presence of IM. PMID- 15790702 TI - IgM colocalises with complement and C reactive protein in infarcted human myocardium. AB - AIMS: Reperfusion of ischaemic myocardium after acute myocardial infarction (AMI) can induce ischaemia/reperfusion (I/R) injury, as a result of local activation of the complement system. C reactive protein (CRP) is involved in this activation. This study analysed the potential role of IgM in complement activation in the infarcted human myocardium. METHODS: Immunochemical analysis was carried out on heart specimens from 59 patients who died from AMI. Serial slides of frozen tissue from the infarction site were stained for IgM, complement factors C3d and C5b-9 (membrane attack complex), and CRP. RESULTS: IgM deposits were found on the plasma membrane, cross striations, and in the cytoplasm of jeopardised cardiomyocytes in infarcts of one to five days duration. IgM depositions were remarkably similar to those of CRP and both complement factors. The relative staining intensities of IgM and CRP varied greatly among patients. CONCLUSIONS: Similar to CRP, IgM targets complement locally to jeopardised cardiomyocytes in the human heart after AMI. Localisation patterns and relative staining intensities suggest that IgM and CRP recognise similar epitopes in the ischaemic heart, but that the relative contribution of each protein to complement activation in the ischaemic myocardium differs among patients. PMID- 15790704 TI - Differences in growth characteristics and elementary body associated cytotoxicity between Chlamydia trachomatis oculogenital serovars D and H and Chlamydia muridarum. AB - AIM: In vitro growth and elementary body (EB) associated cytotoxicity of two Chlamydia trachomatis strains belonging to serovars D and H and C muridarum were compared to identify difference(s) that correlate with virulence variations between these strains in the mouse model of human female genital tract infection, and phenotypic characteristics that could explain human epidemiological data on serovar prevalence and levels of shedding during serovar D and H infection. METHODS: Replication cycle kinetics, inclusion characteristics, and EB associated cytotoxicity were assessed in McCoy cell monolayers using culture, light microscopy, and lactate dehydrogenase release. RESULTS: Over 72 hours, more rapid production and release of inclusion forming units (ifu) allowed C muridarum to initiate two replication rounds, resulting in 4-8 times more ifu/input unit of infection than with serovars D and H. Although C muridarum EBs were significantly more cytotoxic to McCoy cell monolayers than serovar D at moderate and high multiplicity of infection ratios (MOI), serovar H EBs were significantly more cytotoxic than C muridarum, even at the lowest MOI tested. CONCLUSIONS: These phenotypic differences are consistent with the more invasive course and severe pathological outcome of infection in mice infected with C muridarum, providing an objective basis for questioning the appropriateness of C muridarum as a surrogate for the human biovar of C trachomatis in the murine model of female genital tract infection. The differences seen between the human strains could help explain human epidemiological data relating to differences in prevalence and level of shedding that occurs during infection with oculogenital serovars D and H. PMID- 15790703 TI - Genomic instability in radial growth phase melanoma cell lines after ultraviolet irradiation. AB - BACKGROUND/AIMS: Although ultraviolet (UV) irradiation, apoptosis, and genomic instability are all potentially involved in the pathogenesis of melanoma, in vitro studies investigating these changes in the radial growth phase of this neoplasm are still lacking; therefore, this study was designed to investigate these changes. METHOD: An in vitro system consisting of three radial growth phase Wistar melanoma cell lines (WM35, WM3211, and WM1650) was established. Cells were UV irradiated (10 mJ/cm2 for UVB and 6 J/cm2 for UVA), harvested after UV exposure, and evaluated for viability and apoptosis using Trypan blue and terminal deoxynucleotidyl transferase mediated dUTP digoxigenin nick end labelling assays, respectively. Polymerase chain reaction based microsatellite assays were used to examine the cell lines for the presence of microsatellite instability (MSI) using 21 markers at the 1p, 2p, 3p, 4q, 9p, and 17p regions. RESULTS: Exposure to UV initiated progressive cell death associated with pronounced apoptosis, with UVA having a greater effect than UVB. MSI was found in UVB (WM35 and WM3211) and UVA (WM35) irradiated cell lines at 1p, 9p, and 17p, but not in non-irradiated cells. The prevalence of MSI was higher after UVB irradiation (14%) than UVA irradiation (4.7%), and was most frequently found at D1S233. CONCLUSIONS: The ability of erythemogenic UV irradiation to induce both apoptosis and MSI in radial growth phase melanoma cells is suggestive of its role in melanoma pathogenesis. This instability may reflect a hypermutability state, oxidative stress induced DNA damage, replication infidelity, or a combination of these factors. PMID- 15790705 TI - High co-prevalence of genogroup 1 TT virus and human papillomavirus is associated with poor clinical outcome of laryngeal carcinoma. AB - BACKGROUND: The aetiology and factors leading to the progression of laryngeal cancer are still unclear. Although human papillomavirus (HPV) has been suggested to play a role, reports concerning the effect of HPV infection on tumour development are controversial. Recently, transfusion transmitted virus (TTV) was suggested to play a role in certain infections as a causative or coinfecting agent. AIMS: To investigate whether the development and progression of laryngeal squamous cell carcinoma is associated with coinfection with TTV and HPV. METHODS: The prevalence of TTV and HPV was investigated using the polymerase chain reaction in tissue samples from 40 healthy individuals, 10 patients with recurrent papillomatosis, five patients with papillomatosis with malignant transformation, and 25 patients with laryngeal carcinoma. The obtained prevalence data were compared and analysed statistically. RESULTS: In the 11 patients with carcinoma who had metastasis or relapse there was a high rate of coinfection with genogroup 1 TTV and HPV (eight of 11), whereas in the 14 without tumour progression no coinfection was found. Coinfection was associated with significantly lower tumour free survival in patients with carcinoma (p < 0.001). Furthermore, four of five patients who had papillomatosis with malignant transformation were coinfected with genogroup 1 TTV and HPV. CONCLUSIONS: Although the nature of cooperation between HPV and TTV needs to be investigated further, coinfection with genogroup 1 TTV and HPV appears to be associated with poor clinical outcome in laryngeal cancer. PMID- 15790706 TI - Bone marrow biopsy morbidity: review of 2003. AB - BACKGROUND: Although some hazards are recognised, in general, bone marrow aspiration and trephine biopsy are thought to be safe procedures. Until recently, no attempt had been made to quantify any attendant risks. For this reason, documentation of adverse events was begun in 2001, under the auspices of the British Society for Haematology. Three consecutive years have now been surveyed, the results for 2003 being presented here and compared with earlier results. METHODS: Members of the British Society of Haematology were requested to document adverse events associated with diagnostic bone marrow aspirates and trephine biopsies between 1 January and 31 December, 2003. Data were collected early in 2004. RESULTS: In total, 19,259 procedures were reported from 63 hospitals, 13,147 being combined procedures and 6112 aspirates without a trephine biopsy. Sixteen adverse events were reported, representing 0.08% of total reported procedures. The major adverse event was haemorrhage, which comprised 11 of the 16 adverse events. Although infrequent, adverse events were associated with significant morbidity and three were judged as very serious. The major risk factors for haemorrhage, in order of frequency, were diagnosis of a myeloproliferative disorder, aspirin treatment, other putative platelet dysfunctions, and thrombocytopenia. CONCLUSIONS: Adverse events following trephine biopsies and bone marrow aspirates are rare, but nevertheless can have considerable impact on individual patients. PMID- 15790707 TI - Histological features used in the diagnosis of melanoma are frequently found in benign melanocytic naevi. AB - AIMS: The histological features used in the diagnosis of melanoma may be present in benign naevi, but quantitative data are not available. The aim of this study was to establish the real prevalence of such features in naevi. METHODS: Ten dermatopathologists, from nine Italian institutions, studied a series of naevi. Eleven histological parameters currently used in melanoma diagnosis were analysed: asymmetry, poor circumscription, predominance of single melanocytes, irregular confluent nests, suprabasal melanocytes, hair follicle involvement, absence of maturation, cytological atypia, dermal lymphocytic infiltrate, mitoses, and necrosis. RESULTS: Ninety one naevi were examined: 22 junctional, 59 compound, and 10 intradermal. None of the studied parameters was seen in 22 of the benign naevi studied. One or more investigated features were found in 69 naevi. Poor circumscription was found in 49 cases, single melanocytic predominating in 42, asymmetry in 41, irregular confluent nests in 16, cytological atypia in 14, suprabasal melanocytes in seven, and hair follicle involvement in seven; absence of maturation, mitoses and necrosis were not found. CONCLUSIONS: The histological features used for the histological diagnosis of melanoma are often present in benign melanocytic naevi. This suggests a critical, non-mechanical use of them in melanoma diagnosis. PMID- 15790708 TI - Pilot study for the development of a new campylobacter selective medium at 37 degrees C using aztreonam. AB - AIMS: To overcome contamination and temperature inhibition by isolating campylobacter at 37 degrees C. METHODS: The beta lactam antibiotic aztreonam was included in a selective medium because of its inhibitory activity against Gram negative organisms but not against Campylobacter jejuni. Vancomycin and amphotericin were added to inhibit Gram positive bacteria and yeasts. RESULTS: The aztreonam amphotericin vancomycin (AAV) experimental campylobacter selective medium showed growth microaerobically at 37 degrees C of C jejuni, C coli, C lari, C hyointestinalis, C fetus subsp. fetus, and C jejuni subsp. doylei after 24 to 48 hours of incubation. Six campylobacter NCTC strains demonstrated a minimum inhibitory concentration (MIC) > or = 256 mg/litre for vancomycin and aztreonam, whereas C upsaliensis and two "campylobacter-like" strains now reclassified under genus helicobacter--H cinaedi and H fennelliae--had a MIC of 4 mg/litre for vancomycin and aztreonam. In the pilot study (150 samples), AAV medium (37 degrees C) had a higher sensitivity for isolating campylobacters: 14 were isolated on AAV compared with 10 on modified CDA (43 degrees C) over three days, and nine were isolated on AAV medium compared with five on modified CDA (43 degrees C) after 24 hours of incubation. Contamination rates remained low. CONCLUSION: The medium was devised in a pilot study performed between 1990 and 1993; however, this is the first report of AAV medium used as a selective medium capable of growing six campylobacters of pathogenic importance at 37 degrees C. Further studies are indicated. PMID- 15790709 TI - Results of a questionnaire regarding criteria for adequacy of endometrial biopsies. AB - AIMS: Pathologists are faced with increasing numbers of endometrial biopsies containing scant tissue. Anecdotal evidence points to significant variation among pathologists regarding criteria used to assess adequacy, and no standard recommendations exist. An initial audit showing variation in endometrial biopsy adequacy reporting prompted this assessment of the criteria used by specialist gynaecological pathologists for the classification of adequacy. METHODS: A questionnaire regarding criteria used for endometrial biopsy assessment adequacy was sent to members of the British Association of Gynaecological Pathologists and the National Gynaecological Pathology External Quality Assessment Scheme (UK). One hundred and thirty questionnaires were distributed and 61 pathologists responded. RESULTS: The responses showed great variation in criteria used to classify endometrial biopsies as adequate. Most respondents felt it would be useful if criteria were proposed to aid this assessment. CONCLUSIONS: Wide variation exists among specialist gynaecological pathologists regarding what constitutes an adequate endometrial biopsy. The gynaecologist should interpret the biopsy report in the light of clinical, radiological, and hysteroscopic features. The presence of scanty tissue in postmenopausal women with a thin endometrium and no focal lesion is expected, and is not a reason for repeat biopsy. Pathologists should exercise caution before classifying endometrial biopsies as inadequate, because this may have medicolegal and management implications. PMID- 15790711 TI - Rhodococcus equi brain abscess in a patient without HIV. AB - Rhodococcus equi, a Gram positive organism, is a cause of infections in immunocompromised individuals. In humans, it mainly causes disease in those infected with human immunodeficiency virus (HIV), and generally presents as chronic pulmonary infection. It may also cause intracranial infections, which manifest as brain abscesses. This report describes a case of rhodococcus brain and pulmonary infection in a patient who did not have HIV or another disorder of cell mediated immunity. He was treated with intravenous imipenem, vancomycin, and rifampin for eight weeks and recovered from the infection. PMID- 15790710 TI - Strongyloides stercolaris infection mimicking a malignant tumour in a non immunocompromised patient. Diagnosis by bronchoalveolar cytology. AB - Autoinfective strongyloidiasis is often fatal in immunosuppressed patients or in immunocomprised hosts. An interesting case of Strongyloides stercolaris hyperinfection was seen in an immunocompetent patient. This report describes a case of fatal strogyloidiasis in a 79 year old man, who had suffered gastrointestinal discomfort for years, and who presented because of respiratory illness. A chest radiograph showed an irregular mass close to the mediastinum and interstitial infiltrates, but blood eosinophilia was not observed. Cytological examination of the samples obtained from bronchial aspiration and brushing identified several filariform larvae. Thus, cytology was essential for the correct diagnosis in this patient and is a very reliable method to diagnose lung parasitosis. PMID- 15790712 TI - The retroperitoneal surface in distal caecal and proximal ascending colon carcinoma: the Cinderella surgical margin? AB - BACKGROUND: Mesorectal margin tumour involvement is a predictor of local recurrence in rectal carcinoma and an indication for postoperative radiotherapy in suitable patients. However, the prevalence of non-peritonealised surgical margin involvement in ascending colon carcinoma is unknown. AIMS: To test the hypothesis that retroperitoneal surgical margin (RSM) tumour involvement occurs in distal caecal and proximal ascending colon carcinoma. METHODS/RESULTS: One hundred right hemicolectomy specimens, removed for adenocarcinoma of the caecum or proximal ascending colon, were studied. During routine specimen dissection, at least one additional tissue block was taken to include the tumour and the RSM. The tumour distance from the RSM was recorded. RSM tumour involvement was present in seven cases (7%). Direct (non-nodal) RSM tumour involvement (five cases) only occurred in posterior or circumferential tumours. CONCLUSIONS: RSM tumour involvement occurs within a considerable number of distal caecal and proximal ascending colon carcinomas. The rate of RSM tumour involvement identified here is similar to a previously published local recurrence rate of 10% in caecal carcinoma, suggesting that RSM tumour involvement may be a predictor of recurrence in these tumours. Therefore, patients with distal caecal or proximal ascending colon carcinoma and RSM tumour involvement may benefit from postoperative radiotherapy. PMID- 15790713 TI - Immunodetection of SV40 large T antigen in human central nervous system tumours. AB - BACKGROUND/AIMS: DNA sequences from Simian virus 40 (SV40) have been previously isolated from various human tumours of the central nervous system (CNS). This study aimed to investigate a series of tumours of the CNS for the expression of the SV40 large T antigen (Tag), which is an oncogenic protein of the virus. METHODS: A French series of 82 CNS tumours was investigated for Tag expression using a monoclonal antibody and immunohistochemistry. A Tag positive hepatocellular carcinoma cell line from transgenic mice and a kidney biopsy from a patient infected by SV40 were used as positive controls. RESULTS: None of the tumours (20 ependymomas, 20 glioblastomas, 12 oligodendrogliomas, three plexus choroid adenomas, two plexus choroid carcinomas, 15 meningiomas, and 10 medulloblastomas) contained SV40 Tag positive cells. CONCLUSIONS: The lack of SV40 Tag in 82 CNS tumours of various types is at variance with previous studies from different countries, and suggests that the virus may not be an important factor in CNS tumorigenesis, at least in French cases. PMID- 15790714 TI - Candida albicans peritonitis in a patient with Felty's syndrome. AB - A 53 year old man with Felty's syndrome presented with abdominal pain and fever. He underwent a laparotomy after starting broad spectrum antibiotics. An intestinal biopsy showed skip ulcers with fungal hyphae. Peritoneal exudates grew Candida albicans. He was started on intravenous fluconazole and then switched to liposomal amphotericin to which he showed a good clinical response. After one month at home he was readmitted with candidosis and died of a myocardial infarction. PMID- 15790715 TI - Segmental pulmonary veno-occlusive disease secondary to lung cancer. AB - This report describes two cases of segmental pulmonary vein occlusion secondary to lung malignancy in which lung biopsies showed histological features of veno occlusive disease. These are the first cases to be reported in the literature in which such lung parenchymal histological changes are described in association with lung malignancy. PMID- 15790716 TI - Pseudospirochaetosis of the urinary bladder. AB - This report describes an elderly patient with urinary symptoms who showed surface colonisation of the transitional mucosa of the bladder by an unusual haematoxophilic microorganism superficially resembling the "blue fuzz" seen in colonic biopsies showing intestinal spirochaetosis. Special stains showed that the organisms were Gram and Giemsa positive, weakly argyrophilic, and Ziehl Nielsen negative. Immunostains were negative for Helicobacter pylori and electron microscopy revealed curious curved bodies, which were difficult to classify. Therefore, this condition was described as pseudospirochaetosis of the urinary bladder. The urinary symptoms regressed on treatment with ciprofloxacin. The clinicopathological relevance of these findings is discussed in the report. PMID- 15790717 TI - Recurrent severe hyperandrogenism during pregnancy: a case report. AB - This report describes the case of a 28 year old woman with virilisation occurring in two successive pregnancies. Recurrent maternal virilisation is rare (seven previous reports) and this case is unique in its severity. Differential diagnoses include ovarian disease and fetal aromatase deficiency. New techniques to exclude a fetal cause were used in this case. This patient presented during the third trimester of her first pregnancy with rapid onset of hirsuitism, increased musculature, and deepening voice. A blood hormone profile revealed significant hyperandrogenism (testosterone, 72.4 nmol/litre; normal range, 0.5-3.0). She delivered a normal boy and maternal androgen concentrations returned rapidly to normal (testosterone, 0.8 nmol/litre). She presented two years later, during her second pregnancy, with similar symptoms and biochemistry (testosterone, 47.5 nmol/litre). Again, she delivered a healthy normal boy and androgens returned immediately to normal (serum testosterone, 2.0 nmol/litre). Ultrasonography revealed no evidence of ovarian (or adrenal) masses in either pregnancy. Umbilical cord venous blood sampling and placental assays revealed no evidence of fetal aromatase deficiency. Recurrent hyperandrogenism during pregnancy is rare. Ovarian luteoma rarely recurs and hyperreactio luteinalis does not lead to such pronounced androgen concentrations. Therefore, this patient has a unique ovarian condition that could be harmful to offspring and mother. PMID- 15790718 TI - Diagnostic cues for natural killer cell lymphoma: primary nodal presentation and the role of in situ hybridisation for Epstein-Barr virus encoded early small RNA in detecting occult bone marrow involvement. AB - Natural killer (NK) cell lymphomas are rare, and atypical features might lead to diagnostic pitfalls. This report describes an unusual patient in whom lymphoma occurred initially as isolated lymph node involvement, an exceptional presentation of an almost exclusively extranodal disease. Furthermore, during the terminal haemophagocytosis in the bone marrow, lymphoma cells lost the expression of the NK cell marker, CD56, making the histopathological diagnosis of bone marrow involvement difficult. This was resolved by in situ hybridisation for Epstein-Barr virus encoded small RNA, which detected occult bone marrow infiltration. PMID- 15790719 TI - Oncocytic carcinoid tumour of the bladder. PMID- 15790720 TI - Ectopic supernumerary kidney presenting as inguinal hernia. PMID- 15790721 TI - Monckeberg medial calcific sclerosis mimicking malignant calcification pattern at mammography. PMID- 15790722 TI - Endothelial progenitor cells in non-small cell lung cancer. PMID- 15790723 TI - Extramedullary haemopoiesis. PMID- 15790724 TI - Adult-onset growth hormone and insulin-like growth factor I deficiency reduces neoplastic disease, modifies age-related pathology, and increases life span. AB - Disruption of the insulin/IGF-I pathway increases life span in invertebrates. However, effects of decreased IGF-I signaling in mammalian models remain controversial. Using a rodent model with a specific and limited deficiency of GH and IGF-I, we report that GH and IGF-I deficiency throughout life [GH deficiency (GHD)] has no effect on life span compared with normal, heterozygous animals. However, treatment of GHD animals with GH from 4-14 wk of age [adult-onset (AO) GHD] increased median and maximal life span by 14% and 12%, respectively. Analysis of end-of-life pathology indicated that deficiency of these hormones decreased tumor incidence in GHD and AO-GHD animals (18 and 30%, respectively) compared with heterozygous animals and decreased the severity of, and eliminated deaths from, chronic nephropathy. Total disease burden was reduced by 24% in GHD and 16% in AO-GHD animals. Interestingly, the incidence of intracranial hemorrhage increased by 154 and 198% in GHD and AO-GHD animals, respectively, compared with heterozygous animals. Deaths from intracranial hemorrhage in AO-GHD animals were delayed by 14 wk accounting for the increased life span compared with GHD animals. The presence of GH and IGF-I was necessary to maximize reproductive fitness and growth of offspring early in life and to maintain cognitive function and prevent cartilage degeneration later in life. The diverse effects of GH and IGF-I are consistent with a model of antagonistic pleiotropy and suggest that, in response to a deficiency of these hormones, increased life span is derived at the risk of functional impairments and tissue degeneration. PMID- 15790725 TI - Three peroxisome proliferator-activated receptor isotypes from each of two species of marine fish. AB - The cloning and characterization of cDNAs and genes encoding three peroxisome proliferator-activated receptor (PPAR) isotypes from two species of marine fish, the plaice (Pleuronectes platessa) and the gilthead sea bream (Sparus aurata), are reported for the first time. Although differences in the genomic organization of the fish PPAR genes compared with their mammalian counterparts are evident, sequence alignments and phylogenetic comparisons show the fish genes to be homologs of mammalian PPARalpha, PPARbeta/delta, and PPARgamma. Like their mammalian homologs, fish PPARs bind to a variety of natural PPAR response elements (PPREs) present in the promoters of mammalian or piscine genes. In contrast, the mRNA expression pattern of PPARs in the two fish species differs from that observed in other vertebrates. Thus, PPARgamma is expressed more widely in fish tissues than in mammals, whereas PPARalpha and beta are expressed similarly in profile to mammals. Furthermore, nutritional status strongly influences the expression of all three PPAR isotypes in liver, whereas it has no effect on PPAR expression in intestinal and adipose tissues. Fish PPARalpha and beta exhibit an activation profile similar to that of the mammalian PPAR in response to a variety of activators/ligands, whereas PPARgamma is not activated by mammalian PPARgamma-specific ligands. Amino acid residues shown to be critical for ligand binding in mammalian PPARs are not conserved in fish PPARgamma and therefore, together with the distinct tissue expression profile of this receptor, suggest potential differences in the function of PPARgamma in fish compared with mammals. PMID- 15790727 TI - Estrogen and estrogen receptor-{beta} (ER{beta})-selective ligands induce galanin expression within gonadotropin hormone-releasing hormone-immunoreactive neurons in the female rat brain. AB - Among the many factors that integrate the activity of the GnRH neuronal system, estrogens play the most important role. In females, estrogen, in addition to the negative feedback, also exhibits a positive feedback influence upon the activity and output of GnRH neurons to generate the preovulatory LH surge and ovulation. Until recently, the belief has been that the GnRH neurons do not contain estrogen receptors (ERs) and that the action of estrogen upon GnRH neurons is indirect involving several, estrogen-sensitive neurotransmitter and neuromodulator systems that trans-synaptically regulate the activity of the GnRH neurons. Based on our recent findings that GnRH neurons of the female rat coexpress galanin, that galanin is a potent GnRH-releasing peptide, and that ERbeta is present in GnRH neurons, we have evaluated the effect of 17beta-estradiol and two ERbeta selective agonists (WAY-200070, WAY-166818) on the expression of galanin within GnRH neurons. By combining immunocytochemistry for GnRH and in situ hybridization histochemistry for galanin, we demonstrate that 17beta-estradiol (20 mug/kg, sc) stimulates galanin expression within GnRH-immunoreactive neurons in a time dependent manner. A significant increase was observed 2 h after its administration to ovariectomized rats. However, a more robust expression required 3-d treatment regimen. Treatment with the beta-selective ligands resulted in similar observations, although no statistical analysis is available for the 2 hr survival. These observations strongly suggest that estrogen and the ERbeta selective ligands stimulate galanin expression within GnRH neurons via ERbeta, although an indirect mechanism via interneurons still cannot be ruled out. PMID- 15790726 TI - Effects of chronic hyperghrelinemia on puberty onset and pregnancy outcome in the rat. AB - Ghrelin, the endogenous ligand of the GH secretagogue receptor, has been recently involved in a wide array of biological functions, including signaling of energy insufficiency and energy homeostasis. On the basis of the proven reproductive effects of other regulators of energy balance, such as the adipocyte-derived hormone leptin, we hypothesized that systemic ghrelin may participate in the control of key aspects of reproductive function. To test this hypothesis, the effects of daily treatment with ghrelin were assessed in rats, pair-fed with control animals, in two relevant reproductive states, puberty and gestation, which are highly dependent on proper energy stores. Daily sc injection of ghrelin (0.5 nmol/12 h; between postnatal d 33 and 43) significantly decreased serum LH and testosterone levels and partially prevented balano-preputial separation (as an external index of puberty onset) in pubertal male rats. On the contrary, chronic administration of ghrelin to prepubertal females, between postnatal d 23 and 33, failed to induce major changes in serum levels of gonadotropins and estradiol, nor did it modify the timing of puberty, as estimated by the ages at vaginal opening and first estrus. Moreover, females treated with ghrelin at puberty subsequently displayed normal estrous cyclicity and were fertile. Conversely, ghrelin administration (0.5 nmol/12 h) during the first half of pregnancy (d 1-11) resulted in a significant decrease in pregnancy outcome, as estimated by the number of pups born per litter, without changes in the number of successful pregnancies at term or gestational length. Overall, our data indicate that persistently elevated ghrelin levels, as a putative signal for energy insufficiency, may operate as a negative modifier of key reproductive states, such as pregnancy and (male) puberty onset. PMID- 15790728 TI - Extracellular adenosine 5'-triphosphate modulates interleukin-6 production by human thyrocytes through functional purinergic P2 receptors. AB - We investigated the presence of P2 receptors (P2Rs) in human thyrocytes and their possible involvement in the modulation of cytokine release. P2Rs expression was assessed by RT-PCR and, when possible, by immunoblotting. Human primary thyrocytes express the mRNA for the following P2X and P2Y subtypes: P2X(3), P2X(5), P2X(6), P2X(7), and P2Y(1), P2Y(2), P2Y(4), and P2Y(11). Stimulation with extracellular nucleotides of fura-2-loaded thyrocytes triggered an intracellular Ca(2+) signal, suggesting expression of functional receptors. Thyrocytes spontaneously released the proinflammatory cytokine IL-6. The ATP-hydrolyzing enzyme apyrase reduced basal IL-6 release, whereas extracellular ATP dose dependently increased IL-6 secretion. Uridine 5'-triphosphate was also an effective stimulus, whereas benzoyl-ATP was ineffective, suggesting a P2Y- rather than P2X-modulated response. Finally, TSH reduced both the intracellular Ca(2+) ([Ca(2+)](i)) rise and IL-6 release triggered by P2Rs stimulation. In conclusion, we provide functional, pharmacological, and biochemical evidence that human primary thyrocytes express P2YR and P2XR subtypes, coupled to increases in ([Ca(2+)](i)) and secretion of IL-6. P2R-dependent modulation of IL-6 release from human thyrocytes suggests a novel mechanism whereby an inflammatory and/or immune-mediated damage can be initiated and amplified in the thyroid. PMID- 15790729 TI - Role of 14-3-3 eta as a positive regulator of the glucocorticoid receptor transcriptional activation. AB - The glucocorticoid receptor (GR), a member of the nuclear receptor superfamily, mediates the effects of glucocorticoids. It is known that 14-3-3 family proteins interact with GR and regulate its transcriptional activity. They also bind to several molecules and influence many cellular events by altering their subcellular localization and/or acting as a chaperone. Recently, it has been proposed that ligand-activated degradation of GR occurs via the ubiquitin proteasomal degradation pathway and that inhibition of proteasomal activity induces up-regulation of GR and enhances the transcriptional activity of GR. To examine the function of 14-3-3eta in the glucocorticoid-dependent signal pathway, we studied the regulatory role of 14-3-3eta in ligand-induced GR transcriptional activation. 14-3-3eta Enhanced the transcriptional activity of GR, and the levels of GR were higher in cells transfected with the 14-3-3eta expression vector in response to glucocorticoid. The GR level increased in both cytosol and nucleus, and endogenous GR was also elevated by 14-3-3eta in HeLa cells. 14-3-3eta Inhibited ligand-induced down-regulation of GR. Proteasomal inhibition did not induce any synergistic effect on the 14-3-3eta-induced increase in GR in response to glucocorticoid, and inhibition of translation did not block elevation of GR by 14-3-3eta, indicating that 14-3-3eta induces stabilization of GR. These results suggest that 14-3-3eta functions as a positive regulator in the glucocorticoid signal pathway by blocking the degradation of GR and inducing an elevation of GR, thus enhancing the transcriptional activity of GR. PMID- 15790730 TI - Critical role of macrophage migration inhibitory factor activity in experimental autoimmune diabetes. AB - Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine that plays a pivotal role in several immunoinflammatory and autoimmune diseases. In this study we examined the role of MIF in the development of immunoinflammatory diabetes induced in susceptible strains of mice by multiple low doses of streptozotocin. We found that MIF protein was significantly elevated in islet cells during the development of diabetes, and that targeting MIF activity with either neutralizing antibody or the pharmacological inhibitor (S,R)-3-(4 hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester, markedly reduced clinical and histopathological features of the disease, such as hyperglycemia and insulitis. Lymphocytes from mice treated with the MIF inhibitors exhibited reduction of both islet antigen-specific proliferative responses and adhesive cell-cell interactions. Neutralization of MIF also down-regulated the ex vivo secretion of the proinflammatory mediators, TNF-alpha, interferon-gamma, and nitric oxide, while augmenting that of the antiinflammatory cytokine, IL-10. This study provides the first in vivo evidence for a critical role for MIF in the immune-mediated beta-cell destruction in an animal model of human type 1 diabetes mellitus and identifies a new therapeutic strategy for the prevention and treatment of this disease in humans that is based on the selective inhibition of MIF activity. PMID- 15790731 TI - Estrogen modulates endothelial and neuronal nitric oxide synthase expression via an estrogen receptor beta-dependent mechanism in hypothalamic slice cultures. AB - Although it is evident that estrogen has important physiological effects in the brain, the signaling mechanisms mediating these effects remain unclear. We recently showed that estrogen mediates attenuated blood pressure responses to psychological stress in ovariectomized female rats through brain nitric oxide (NO). An area likely to mediate these effects is the hypothalamic paraventricular nucleus (PVN), because here NO exerts inhibitory effects on autonomic output to the periphery. Because little is known about how estrogen acts on the NO system in the PVN, our aim was to study the effects of estrogen on the NO system in the PVN of hypothalamic slices cultures. We show that 17beta-estradiol (E2; 1 nm) increases endothelial NO synthase (eNOS) protein expression and decreases the numbers of neuronal NOS (nNOS)-positive neurons in the PVN after 8 and 24 h, respectively. Using the nonselective estrogen receptor (ER) antagonist, ICI 182,780 (10 nm), we determined that E2-induced changes in NOS expression in the PVN are ER dependent. Using the ERbeta agonist, genistein (0.1 microm), we determined that activation of ERbeta induces increased eNOS expression and a decreased number of nNOS-positive neurons. We used the selective ERalpha agonist, propyl-pyrazole-triol (10 nm), and antagonist, methyl-piperidino-pyrazole (1 microm), to exclude the possibility that ERalpha is involved in the E2-induced increase in eNOS and nNOS in the PVN. These results demonstrate that E2 induces changes in NOS expression in the PVN and that these effects are ERbeta dependent. PMID- 15790732 TI - Vesiculo-erosive oral mucosal disease--management with topical corticosteroids: (1) Fundamental principles and specific agents available. AB - Vesiculo-erosive diseases of the oral mucosa pose a major challenge in oral medicine, because they are chronic, painful, and interfere with the daily activities and quality of life of the patients, including disturbing eating, drinking, talking, and personal relationships. Many are autoimmune diseases, and corticosteroid therapy is currently central to their treatment. These diseases present with inflammation and alterations to epithelial integrity, through cell and/or humoral immunity-mediated attack on epithelial-connective tissue targets. Until recently, despite their serious adverse effects, it was necessary to prescribe systemic corticosteroids to control severe erosive oral diseases. Now, however, many of these diseases can be controlled by high-potency topical corticosteroids, which have proved to be highly efficacious and to cause fewer adverse effects compared with systemic corticosteroids. Nevertheless, although topical corticosteroids are still the most widely used drugs in the practice of oral medicine, the scientific body of evidence for their use in the oral cavity is virtually non-existent, and therefore many of the protocols followed are, of necessity, drawn from experience of their use in a dermatological setting. This review aims to set out the key aspects of the use of topical corticosteroids in oral medicine. The issues covered include the indications and basic rules for their use, the types of corticosteroids, the drug selection, and the specific formulations. PMID- 15790733 TI - Vesiculo-erosive oral mucosal disease--management with topical corticosteroids: (2) Protocols, monitoring of effects and adverse reactions, and the future. AB - Although topical corticosteroids (TCs) are the most widely used drugs in oral medicine, and specifically in the treatment of vesiculo-erosive oral mucosal disease, there are few evidence-based data for the correct use of these drugs. In this review, we outline the most widely used protocols, the most common reasons for treatment failure, and the adverse effects documented in the literature. PMID- 15790734 TI - Distinct patterns of angiogenesis in oral and skin wounds. AB - Clinical observation suggests that oral mucosal wounds heal faster than skin; however, little is known about the site-specific differences. Since fetal skin wounds heal rapidly, but are less vascular than adult wounds, we hypothesized that less robust wound angiogenesis might be observed in healing oral mucosa. This study investigated angiogenesis in equivalent-size oral and skin murine wounds. Change in wound bed vascularity was significantly lower in oral wounds than in skin. Also, vascular endothelial growth factor (VEGF) levels were less in oral than cutaneous wounds. Because keratinocytes are a prominent source of VEGF in wounds, we compared VEGF production by oral and epidermal keratinocytes in vitro. Significantly higher levels of VEGF protein and mRNA were observed in epidermal keratinocytes than in oral keratinocytes after 18 hrs of hypoxia. This study demonstrates distinct angiogenesis patterns in oral and skin wounds and intrinsic site-specific differences in VEGF production by keratinocytes. PMID- 15790735 TI - Localization of am3 using EL congenic mouse strains. AB - EL/Sea mice have 100% incidence of the absence of third molars (M3). Our previous linkage analysis using EL/Sea and MSM/Msf mouse strains provides statistical evidence of a major locus for the absence of M3, designated am3, of EL/Sea at the middle region of chromosome 3. To obtain independent evidence for linkage and more precisely determine the location of the am3 locus, we generated EL/Sea congenic strains for am3 in which the restricted interval of chromosome 3 of EL/Sea was replaced by an MSM/Msf-derived homologue. EL/Sea congenic mice that were either heterozygous or homozygous for the MSM/Msf-derived interval exhibited a significant decrease in the incidence of the absence of third molars, confirming previous genome scan results. These results confine the am3 locus to an approximately 4.4-cM region, and demonstrate that other unmapped genes are also involved in the absence of M3 in EL/Sea mice. PMID- 15790737 TI - Inflammation is more persistent in type 1 diabetic mice. AB - Whether diabetes enhances or diminishes the host response to bacteria has been controversial. To determine how diabetes alters the inflammatory response, we inoculated P. gingivalis into the scalps of mice rendered diabetic with multiple low-dose streptozotocin treatment. On day 1, a moderate to severe inflammatory infiltrate was noted in both the diabetic and normoglycemic mice. After 3 days, the inflammatory infiltrate was significantly higher in the diabetic compared with the control group (P < 0.05). The mRNA expression of chemokines macrophage inflammatory protein-2 and monocyte chemoattractant protein-1 was strongly and similarly induced 3 hrs and 1 day post-inoculation. By day 3, the levels were reduced in normoglycemic mice but remained significantly higher in the diabetic group (P < 0.05). To determine whether persistent inflammation was specific for the streptozotocin-induced diabetic model, we directly compared the expression of TNF-alpha in streptozotocin-induced and db/db diabetic mice, which developed type 2 diabetes. Both exhibited prolonged TNF-alpha expression compared with controls. These results suggest that diabetes alters bacteria-host interactions by prolonging the inflammatory response. PMID- 15790736 TI - Modulation of gingival fibroblast minocycline accumulation by biological mediators. AB - Gingival fibroblasts actively accumulate tetracyclines, thereby enhancing their redistribution from blood to gingiva. Since growth factors and pro-inflammatory cytokines regulate many fibroblast activities, they could potentially enhance fibroblast minocycline accumulation. To test this hypothesis, we treated gingival fibroblast monolayers for 1 or 6 hours with platelet-derived growth factor-BB (PDGF), fibroblast growth factor-2 (FGF), transforming growth factor-beta1 (TGF), or tumor necrosis factor-alpha (TNF). Minocycline uptake was assayed at 37 degrees by a fluorescence method. All 4 factors significantly enhanced minocycline uptake (P < or = 0.008, ANOVA), primarily by increasing the affinity of transport. Treatment for 6 hours with 10 ng/mL FGF, PDGF, TGF, or TNF enhanced fibroblast minocycline uptake by 19% to 25%. Phorbol myristate acetate enhanced fibroblast minocycline uptake by 28%, suggesting that protein kinase C plays a role in up-regulating transport. These effects on transport provide a mechanism by which systemic tetracyclines could be preferentially distributed to gingival wound or inflammatory sites. PMID- 15790738 TI - Parathyroid-hormone-related protein induces expression of receptor activator of NF-{kappa}B ligand in human periodontal ligament cells via a cAMP/protein kinase A-independent pathway. AB - Periodontal ligament (PDL) cells play important roles in root resorption of human deciduous teeth by odontoclasts (osteoclast-like cells). However, it is unclear how PDL cells regulate osteoclastogenesis. We examined the effects of PTHrP, TGF beta, and EGF, which are all secreted by the tooth germ, on tartrate-resistant acid-phosphatase-positive (TRAP+) cell formation using co-cultures of human PDL cells and mouse spleen cells. Only PTHrP promoted TRAP+ cell formation in co cultures. PTHrP induced receptor activator of NF-kappaB ligand (RANKL) mRNA expression and slightly reduced osteoprotegerin (OPG) expression in PDL cells. The cAMP/PKA inhibitors Rp-cAMP, H89, and PKI did not affect PTHrP-induced TRAP+ cell formation. The PKC inhibitor, Ro-32-0432, suppressed RANKL expression in PDL cells and PTHrP-induced TRAP+ cell formation. However, this inhibitor directly modulated the number of osteoclast precursors. Thus, PTHrP induces osteoclastogenesis by increasing the relative expression level of RANKL vs. OPG in PDL cells via a cAMP/PKA-independent pathway. ABBREVIATIONS: PTHrP, parathyroid-hormone-related protein; TGF-beta, transforming growth factor-beta; EGF, epidermal growth factor; RANKL, receptor activator of NF-kappaB ligand; OPG, osteoprotegerin; PDL, periodontal ligament; TRAP, tartrate-resistant acid phosphatase; PKA, protein kinase A; PKC, protein kinase C; MAP, mitogen-activated protein; ERK, extracellular signal-regulated kinase; cAMP, cyclic Adenosine 3'5' Monophosphate. PMID- 15790739 TI - Painful tooth stimulation elevates matrix metalloproteinase-8 levels locally in human gingival crevicular fluid. AB - Recent studies have demonstrated that pulpal pain can induce neurogenic inflammatory reactions in gingiva and the expression of pro-inflammatory neuropeptides in gingival crevicular fluid (GCF). Neuropeptides co-ordinate the activity of immuno-effector cells and may influence the secretion of matrix metalloproteinase (MMP)-8, the major tissue-destructive protease in GCF. With this background, we studied whether experimental pulpal pain can trigger changes in GCF MMP-8 levels. The molecular forms of MMP-8 in the GCF of stimulated and non-stimulated teeth were analyzed by Western immunoblot, and MMP-8 levels by quantitative immunofluorometric assay. Painful stimulation of the upper incisor provoked significant elevations in GCF MMP-8 levels of the stimulated tooth. Western immunoblot revealed elevations in both neutrophil- and mesenchymal-type MMP-8 isoforms. At the same time, the GCF MMP-8 levels of the non-stimulated teeth were not changed. Analysis of these data indicated that pulpal pain can induce local elevations in MMP-8 levels in GCF. PMID- 15790740 TI - Initial subgingival colonization of 'pristine' pockets. AB - The treatment of periodontitis/peri-implantitis involves the reduction/eradication of periopathogens. After therapy, beneficial and pathogenic species recolonize the subgingival area. The dynamics of recolonization and especially the role of the supragingival environment in this process are still not well-understood. This prospective, split-mouth study followed the early colonization of 'pristine' pockets created during implant surgery (16 partially edentulous patients), to record the time needed before a complex subgingival flora could be established with the supragingival area as the single source. Four subgingival plaque samples were taken from shallow and medium pockets around implants (test), and neighboring teeth (undisturbed microbiota as reference) 1, 2, and 4 wks after abutment connection. Checkerboard DNA-DNA hybridization and culture data revealed a complex microbiota (including several pathogenic species) in the pristine pockets within a wk, with a minimal increase in counts up to 4 wks. Analysis of these data demonstrated that, even with the supragingival environment as the single source for colonizing bacteria, a complex subgingival microbiota can develop within 1 wk. PMID- 15790741 TI - Introduction to cumulative meta-analysis in dentistry: lessons learned from undertaking a cumulative meta-analysis in periodontology. AB - Improving health and well-being from the consideration of isolated studies is problematic. Systematic reviews have been developed to address this problem and may include a quantitative data synthesis in the form of a meta-analysis, or a cumulative meta-analysis. The value of systematic reviews depends greatly on the availability and quality of the results of primary research. The objective of the current project was to demonstrate the technique of cumulative meta-analysis in dentistry using data from a previously published systematic review. The process highlights an issue that some trials could not be synthesized due to the lack of reporting of measures of variation. This represents a potential source of bias. Investigators are encouraged to consider their trials as part of an information continuum and to report sufficient detail to permit the trials' incorporation into subsequent syntheses. PMID- 15790742 TI - Continuing etching of an all-in-one adhesive in wet dentin tubules. AB - Self-etch adhesives that etch and prime simultaneously are becoming more acidic. We hypothesized that the degree of acidic monomer conversion at the interface and within the tubules was high enough that the acidic reaction would be very self limiting. Dentin surfaces prepared from extracted, unerupted human third molars were treated with Prompt L-Pop (3M ESPE). The prepared teeth were stored in normal saline, and specimens retrieved at intervals < or = 4 wks were randomly selected for light, scanning electron microscopic and micro-Raman spectroscopic analysis. Morphologic and spectroscopic analyses indicated dentin demineralization and adhesive penetration throughout the demineralized layer and tubules. Increased dentin demineralization and loss of adhesive integrity were noted after aqueous storage. The degree of monomer conversion at the interface was consistently greater than conversion within the tubules. Fluid within the tubules may inhibit monomer conversion. The acidic characteristics of this adhesive may be retained and, thus, continue to affect/demineralize the surrounding dentin. PMID- 15790743 TI - Ultrastructural correlates of in vivo/in vitro bond degradation in self-etch adhesives. AB - The morphologic correlates of bond degradation in self-etching primers have not been fully elucidated. We hypothesized that there is no difference between the mechanism of degradation of self-etching primers in vivo and in vitro. Class I cavities prepared in vivo in 24 caries-free human molars were bonded with Clearfil SE Bond or Clearfil Protect Bond, and restored with resin composites. Eight teeth were extracted after 24 hrs, and the rest after 1 yr. The same protocol was repeated in vitro with extracted molars. Degradation of resin-dentin bonds was assessed by microtensile bond testing and TEM of interfaces after tracer immersion. Both in vivo and in vitro bond strengths decreased with time for SE Bond but not for Protect Bond, with more pronounced water treeing observed in the former adhesive under both aging conditions. There is no difference between the mechanism of degradation of self-etch adhesives in vivo or in vitro. PMID- 15790744 TI - Isocyanato- and methacryloxysilanes promote Bis-GMA adhesion to titanium. AB - In dentistry, adhesion promotion with 3-methacryloyloxypropyltrimethoxysilane is usually sufficient, but its hydrolytic stability is a continuous concern. The hydrolytic stability of an alternative, 3-isocyanatopropyltriethoxysilane, was compared with that of conventional 3-methacryloyloxypropyltrimethoxysilane. Two silanes, both in 0.1 and 1.0 vol-% in ethanol-water, were evaluated in the attachment of an experimental bis-phenol-A-diglycidyldimethacrylate (Bis-GMA) resin to grit-blasted (with two different systems) titanium. Silane hydrolysis was monitored by FTIR spectrometry. Bis-GMA resin was applied and photo polymerized on titanium. The specimens were thermocycled (6000 cycles, 5-55 degrees C). Surface analysis was carried out with scanning electron microscopy. Statistical analysis (ANOVA) showed that the highest shear bond was achieved with 0.1% 3-isocyanatopropyltriethoxysilane (12.5 MPa) with silica-coating, and the lowest with 1.0% 3-methacryloyloxypropyltrimethoxysilane (3.4 MPa) with alumina coating. The silane, its concentration, and the grit-blasting method significantly affected the shear bond strength (p < 0.05). SEM images indicated cohesive failure of bonding, and, in conclusion, 3 isocyanatopropyltriethoxysilane is a potential coupling agent. PMID- 15790745 TI - Discontinuous fiber-reinforced composites above critical length. AB - Micromechanical physics of critical fiber length, describing a minimum filament distance for resin impregnation and stress transfer, has not yet been applied in dental science. As a test of the hypothesis that 9-micron-diameter, 3-mm-long quartz fibers would increase mechanical strength over particulate-filled composites, photocure-resin-pre-impregnated discontinuous reinforcement was incorporated at 35 wt% into 3M Corporation Z100, Kerr Corporation HerculiteXRV, and an experimental photocure paste with increased radiopaque particulate. Fully articulated four-point bend testing per ASTM C 1161-94 for advanced ceramics and Izod impact testing according to a modified unnotched ASTM D 256-00 specification were then performed. All photocure-fiber-reinforced composites demonstrated significant improvements over particulate-filled compounds (p < 0.001) for flexural strength, modulus, work of fracture, strain at maximum load, and Izod toughness, with one exception for the moduli of Z100 and the experimental reinforced paste. The results indicate that inclusion of pre-impregnated fibers above the critical aspect ratio yields major advancements regarding the mechanical properties tested. PMID- 15790746 TI - Dentin erosion simulation by cantilever beam fatigue and pH change. AB - Exposed root surfaces frequently exhibit non-carious notches representing material loss by abrasion, erosion, and/or abfraction. Although a contribution from mechanical stress is often mentioned, no definitive proof exists of a cause effect relationship. To address this, we examined dimensional changes in dentin subjected to cyclic fatigue in two different pH environments. Human dentin cantilever-beams were fatigued under load control in pH = 6 (n = 13) or pH = 7 (n = 13) buffer, with a load ratio (R = minimum load/maximum load) of 0.1 and frequency of 2 Hz, and stresses between 5.5 and 55 MPa. Material loss was measured at high- and low-stress locations before and after cycling. Of the 23 beams, 7 withstood 1,000,000 cycles; others cracked earlier. Mean material loss in high-stress areas was greater than in low-stress areas, and losses were greater at pH = 6 than at pH = 7, suggesting that mechanical stress and lower pH both accelerate erosion of dentin surfaces. PMID- 15790747 TI - Viscoelasticity of dental tissue conditioners during the sol-gel transition. AB - Formation of tissue conditioners is a process of polymer chain entanglements. This study evaluated the influence of composition and structure on dynamic viscoelasticity of concentrated polymer solutions based on poly(ethyl methacrylate) (PEMA) used as tissue conditioners through the sol-gel transition. The hypothesis was that the ethanol content is the most influential factor in determining gelation speed. Rheological parameters were determined with the use of a controlled-stress rheometer. Analysis of variance by orthogonal array L(16)(4(5)) indicated that the strong polar bonding of ethanol (contribution ratio rho = 53.8%; confirming the hypothesis) and molecular weight of polymer powders (rho = 26.7%) had a greater influence on the gelation times of PEMA-based systems than did the molar volume of plasticizers (rho = 9.0%) and concentration of polymers (i.e., powder/liquid ratio) (rho = 4.5%). The results suggest that the gelation of tissue conditioners based on PEMA can be controlled over a wide range by varying the polymer molecular weight, and especially ethanol content. PMID- 15790748 TI - Effect of testosterone on the female anterior cruciate ligament. AB - Injuries to the anterior cruciate ligament (ACL) result in immediate and long term morbidity and expense. Young women are more likely to sustain ACL injuries than men who participate in similar athletic and military activities. Although significant attention has focused on the role that female sex hormones may play in this disparity, it is still unclear whether the female ACL also responds to androgens. The purpose of this study was to determine whether the female ACL was an androgen-responsive tissue. To identify and localize androgen receptors in the female ACL, we used Western blotting and immunofluorescent labeling, respectively, of ACL tissue harvested during surgery from young women (n = 3). We then measured ACL stiffness and assessed total testosterone (T) and free [free androgen index (FAI)] testosterone concentrations, as well as relative estradiol to testosterone ratios (E(2)/T and E(2)/FAI) at three consecutive menstrual stages (n = 20). There were significant rank-order correlations between T (0.48, P = 0.031), FAI (0.44, P = 0.053), E(2)/T (-0.71, P < 0.001), E(2)/FAI (-0.63, P = 0.003), and ACL stiffness near ovulation. With the influences of the other variables controlled, there were significant negative partial rank-order correlations between ACL stiffness and E(2)/T (-0.72, P < 0.001) and E(2)/FAI ( 0.59, P = 0.012). The partial order residuals for T and FAI were not significant. These findings suggest that the female ACL is an androgen-responsive tissue but that T and FAI are not independent predictors of ACL stiffness near ovulation. Instead, the relationship between T, FAI, and ACL stiffness was likely influenced by another hormone or sex hormone binding globulin. PMID- 15790749 TI - Late-night presentation of an auditory stimulus phase delays human circadian rhythms. AB - Although light is considered the primary entrainer of circadian rhythms in humans, nonphotic stimuli, including exercise and melatonin also phase shift the biological clock. Furthermore, in birds and nonhuman mammals, auditory stimuli are effective zeitgebers. This study investigated whether a nonphotic auditory stimulus phase shifts human circadian rhythms. Ten subjects (5 men and 5 women, ages 18-72, mean age +/- SD, 44.7 +/- 21.4 yr) completed two 4-day laboratory sessions in constant dim light (<20 lux). They received two consecutive presentations of either a 2-h auditory or control stimulus from 0100 to 0300 on the second and third nights (presentation order of the stimulus and control was counterbalanced). Core body temperature (CBT) was collected and stored in 2-min bins throughout the study and salivary melatonin was obtained every 30 min from 1900 to 2330 on the baseline and poststimulus/postcontrol nights. Circadian phase of dim light melatonin onset (DLMO) and of CBT minimum, before and after auditory or control presentation was assessed. The auditory stimulus produced significantly larger phase delays of the circadian melatonin (mean +/- SD, -0.89 +/- 0.40 h vs. -0.27 +/- 0.16 h) and CBT (-1.16 +/- 0.69 h vs. -0.44 +/- 0.27 h) rhythms than the control. Phase changes for the two circadian rhythms also positively correlated, indicating direct effects on the biological clock. In addition, the auditory stimulus significantly decreased fatigue compared with the control. This study is the first demonstration of an auditory stimulus phase shifting circadian rhythms in humans, with shifts similar in size and direction to those of other nonphotic stimuli presented during the early subjective night. This novel stimulus may be a useful countermeasure to facilitate circadian adaptation after transmeridian travel or shift work. PMID- 15790751 TI - Depressed Na+-K+-ATPase activity in skeletal muscle at fatigue is correlated with increased Na+-K+-ATPase mRNA expression following intense exercise. AB - We investigated whether depressed muscle Na(+)-K(+)-ATPase activity with exercise reflected a loss of Na(+)-K(+)-ATPase units, the time course of its recovery postexercise, and whether this depressed activity was related to increased Na(+) K(+)-ATPase isoform gene expression. Fifteen subjects performed fatiguing, knee extensor exercise at approximately 40% maximal work output per contraction. A vastus lateralis muscle biopsy was taken at rest, fatigue, 3 h, and 24 h postexercise and analyzed for maximal Na(+)-K(+)-ATPase activity via 3-O methylfluorescein phosphatase (3-O-MFPase) activity, Na(+)-K(+)-ATPase content via [(3)H]ouabain binding sites, and Na(+)-K(+)-ATPase alpha(1)-, alpha(2)-, alpha(3)-, beta(1)-, beta(2)- and beta(3)-isoform mRNA expression by real-time RT PCR. Exercise [352 (SD 267) s] did not affect [(3)H]ouabain binding sites but decreased 3-O-MFPase activity by 10.7 (SD 8)% (P < 0.05), which had recovered by 3 h postexercise, without further change at 24 h. Exercise elevated alpha(1) isoform mRNA by 1.5-fold at fatigue (P < 0.05). This increase was inversely correlated with the percent change in 3-O-MFPase activity from rest to fatigue (%Delta3-O-MFPase(rest-fatigue)) (r = -0.60, P < 0.05). The average postexercise (fatigue, 3 h, 24 h) alpha(1)-isoform mRNA was increased 1.4-fold (P < 0.05) and approached a significant inverse correlation with %Delta3-O-MFPase(rest-fatigue) (r = -0.56, P = 0.08). Exercise elevated alpha(2)-isoform mRNA at fatigue 2.5 fold (P < 0.05), which was inversely correlated with %Delta3-O-MFPase(rest fatigue) (r = -0.60, P = 0.05). The average postexercise alpha(2)-isoform mRNA was increased 2.2-fold (P < 0.05) and was inversely correlated with the %Delta3-O MFPase(rest-fatigue) (r = -0.68, P < 0.05). Nonsignificant correlations were found between %Delta3-O-MFPase(rest-fatigue) and other isoforms. Thus acute exercise transiently decreased Na(+)-K(+)-ATPase activity, which was correlated with increased Na(+)-K(+)-ATPase gene expression. This suggests a possible signal transduction role for depressed muscle Na(+)-K(+)-ATPase activity with exercise. PMID- 15790750 TI - Flash photolysis of caged nitric oxide inhibits proximal tubular fluid reabsorption in free-flow nephron. AB - A nonobstructing optical method was developed to measure proximal tubular fluid reabsorption in rat nephron at 0.25 Hz. The effects of uncaging luminal nitric oxide (NO) on proximal tubular reabsorption were investigated with this method. Proximal fluid reabsorption rate was calculated as the difference of tubular flow measured simultaneously at two locations (0.8-1.8 mm apart) along a convoluted proximal tubule. Tubular flow was estimated on the basis of the propagating velocity of fluorescent dextran pulses in the lumen. Changes in local tubular flow induced by intratubular perfusion were detected simultaneously along the proximal tubule, indicating that local tubular flow can be monitored in multiple sites along a tubule. The estimated tubular reabsorption rate was 5.52 +/- 0.38 nl.min(-1).mm(-1) (n = 20). Flash photolysis of luminal caged NO (potassium nitrosylpentachlororuthenate) was induced with a 30-Hz UV nitrogen-pulsed laser. Release of NO from caged NO into the proximal tubule was confirmed by monitoring intracellular NO concentration using a cell-permeant NO-sensitive fluorescent dye (DAF-FM). Emission of DAF-FM was proportional to the number of laser pulses used for uncaging. Photolysis of luminal caged NO induced a dose-dependent inhibition of proximal tubular reabsorption without activating tubuloglomerular feedback, whereas uncaging of intracellular cGMP in the proximal tubule decreased tubular flow. Coupling of this novel method to measure reabsorption with photolysis of caged signaling molecules provides a new paradigm to study tubular reabsorption with ambient tubular flow. PMID- 15790753 TI - Repeated pregnancies (multiparity) increases venous tone and reduces compliance. AB - In humans, multiparity (repeated pregnancy) is associated with increased risk of cardiovascular disease. In rats, multiparity increases the pressor response to phenylephrine and to acute stress, due in part to changes in tone of the splanchnic arterial vasculature. Given that the venous system also changes during pregnancy, we studied the effects of multiparity on venous tone and compliance. Cardiovascular responses to volume loading (2 ml/100 g body wt), and mean circulatory filling pressure (MCFP, an index of venomotor tone) were measured in conscious, repeatedly bred (RB), and age-matched virgin rats. In addition, passive compliance and venous reactivity of isolated mesenteric veins were measured by pressure myography. There was a greater increase in mean arterial pressure after volume loading in RB rats (+7.2 +/- 2.5 mmHg, n = 8) than virgin rats (-1.4 +/- 1.7 mmHg, n = 7) (P < 0.05). The increase in MCFP in response to norepinephrine (NE) was also greater in RB rats [half maximal effective dose (ED(50)) 3.1 +/- 0.5 nmol.kg(-1).min(-1), n = 6] than virgins (ED(50): 12.1 +/- 2.7 nmol.kg(-1).min(-1), n = 6) (P < 0.05). Pressure-induced changes in passive diameter were lower in isolated mesenteric veins from RB rats (29.3 +/- 1.8 microm/mmHg, n = 6) than from virgins (36.9 +/- 1.3 microm/mmHg, n = 6) (P < 0.05). Venous reactivity to NE in isolated veins was also greater in RB rats (EC(50): 2.68 +/- 0.37x10(-8) M, n = 5) than virgins (EC(50): 4.67 +/- 0.93 x 10( 8) M, n = 8). We conclude that repeated pregnancy induces a long-term reduction in splanchnic venous compliance and augments splanchnic venous reactivity and sympathetic tonic control of total body venous tone. This compromises the ability of the capacitance (venous) system to accommodate volume overloads and to buffer changes in cardiac preload. PMID- 15790752 TI - Cardiovascular response to a group III mGluR agonist in NTS requires NMDA receptors. AB - Previous studies have demonstrated that microinjection of the putative group III metabotropic glutamate receptor (mGluR) agonist, l(+)-2-amino-4-phosphonobutyric acid (L-AP4), into the nucleus tractus solitarius (NTS) produces depressor and sympathoinhibitory responses. These responses are significantly attenuated by a group III mGluR antagonist and may involve ionotropic glutamatergic transmission. Alternatively, a previous report in vitro suggests that preparations of L-AP4 may nonspecifically activate NMDA channels due to glycine contamination (Contractor A, Gereau RW, Green T, and Heinemann SF. Proc Natl Acad Sci USA 95: 8969-8974, 1998). Therefore, the present study tested whether responses to L-AP4 specifically require the N-methyl-D-aspartate (NMDA) receptor and whether they are due to actions at the glycine site on the NMDA channel. To test these possibilities in vivo, we performed unilateral microinjections of L-AP4, glycine, and selective antagonists into the NTS of urethane-anesthetized rats. L-AP4 (10 mM, 30 nl) produced sympathoinhibitory responses that were abolished by the NMDA receptor antagonist 2-amino-5-phosphonovaleric acid (AP-5, 10 mM) but were unaffected by the non-NMDA antagonist 6-nitro-7-sulfamobenzoquinoxaline-2,3-dione (NBQX, 2 mM). Microinjection of glycine (0.02-20 mM) failed to mimic sympathoinhibitory responses to L-AP4, even in the presence of the inhibitory glycine antagonist, strychnine (3 mM). Strychnine blocked pressor and sympathoexcitatory actions of glycine (20 mM) but failed to reveal a sympathoinhibitory component due to presumed activation of NMDA receptors. The results of these experiments suggest that responses to L-AP4 require NMDA receptors and are independent of non-NMDA receptors. Furthermore, although it is possible that glycine contamination or other nonspecific actions are responsible for the sympathoinhibitory actions of L-AP4, our data and data in the literature argue against this possibility. Thus we conclude that responses to L-AP4 in the NTS are mediated by an interaction between group III mGluRs and NMDA receptors. Finally, we also caution that nonselective actions of L-AP4 should be considered in future studies. PMID- 15790754 TI - Use of low concentrations of human IgA anti-tissue transglutaminase to rule out selective IgA deficiency in patients with suspected celiac disease. PMID- 15790755 TI - Identification of ghrelin in human saliva: production by the salivary glands and potential role in proliferation of oral keratinocytes. AB - BACKGROUND: We investigated whether ghrelin is present in human saliva, is produced by salivary glands, and physiologic consequences of these findings. METHODS: Expression of ghrelin and specific receptor mRNA was determined by PCR. Proteins were identified by immunoblotting and size-exclusion fast protein liquid chromatography (FPLC) with consecutive RIA. Specific RIAs were used for quantification of salivary total and bioactive ghrelin. Distribution of ghrelin was investigated by immunohistochemistry in cryosections of the salivary glands. The effect of ghrelin on incorporation of 5-bromo-2'-deoxyuridine as a measure of cell proliferation was investigated in primary oral keratinocytes. RESULTS: Ghrelin is produced by the salivary glands. The hormone was identified in saliva and glands by immunoblotting and by FPLC fractionation of saliva. Immunohistochemistry demonstrated ghrelin distribution in the salivary glands. The receptor was also produced by the glands and by oral keratinocytes and was shown to be functional. Comparison of total ghrelin values for healthy individuals (body mass index, 18-27 kg/m2) showed significantly lower concentrations in saliva than in serum (P < 0.01). The correlation between both matrices was r2 = 0.56 (P < 0.001) with a negative correlation to body mass index (r2 = 0.314; P < 0.01). Bioactive acylated ghrelin was also present in saliva. Incubation of keratinocytes with ghrelin led to significantly increased cell proliferation (P < 0.001). This effect could be completely suppressed by co incubation with NOX-B11 (50 nmol/L), a novel specific inhibitor of acylated ghrelin. CONCLUSIONS: Ghrelin in saliva is produced and released by salivary glands. The effect of ghrelin on oral cell proliferation adds to the pro proliferative action of other salivary growth factors. PMID- 15790756 TI - Liver disease with altered bile acid transport in Niemann-Pick C mice on a high fat, 1% cholesterol diet. AB - Cholestatic hepatitis is frequently found in Niemann-Pick C (NPC) disease. We studied the influence of diet and the low density lipoprotein receptor (LDLR, Ldlr in mice, known to be the source of most of the stored cholesterol) on liver disease in the mouse model of NPC. Npc1-/- mice of both sexes, with or without the Ldlr knockout, were fed a 18% fat, 1% cholesterol ("high-fat") diet and were evaluated by chemical and histological methods. Bile acid transporters [multidrug resistance protein (Mrps) 1-5; Ntcp, Bsep, and OatP1, 2, and 4] were quantitated by real-time RT-PCR. Many mice died prematurely (within 6 wk) with hepatomegaly. Histopathology showed an increase in macrophage and hepatocyte lipids independent of Ldlr genotype. Non-zone-dependent diffuse fibrosis was found in the surviving mice. Serum alanine aminotransferase was elevated in Npc1-/- mice on the regular diet and frequently became markedly elevated with the high-fat diet. Serum cholesterol was increased in the controls but not the Npc1-/- mice on the high fat diet; it was massively increased in the Ldlr-/- mice. Esterified cholesterol was greatly increased by the high-fat diet, independent of Ldlr genotype. gamma Glutamyltransferase was also elevated in Npc1-/- mice, more so on the high-fat diet. Mrps 1-5 were elevated in Npc1-/- liver and became more elevated with the high-fat diet; Ntcp, Bsep, and OatP2 were elevated in Npc1-/- liver and were suppressed by the high-fat diet. In conclusion, Npc1-/- mice on a high-fat diet provide an animal model of NPC cholestatic hepatitis and indicate a role for altered bile acid transport in its pathogenesis. PMID- 15790757 TI - Endocytosis of cholera toxin by human enterocytes is developmentally regulated. AB - Many secretory diarrheas including cholera are more prevalent and fulminant in young infants than in older children and adults. Cholera toxin (CT) elicits a cAMP-dependent chloride secretory response in intestinal epithelia, which accounts for the fundamental pathogenesis of this toxigenic diarrhea. We have previously reported that the action of this bacterial enterotoxin is excessive in immature enterocytes and under developmental regulation. In this study, we tested the hypothesis that enhanced endocytosis by immature human enterocytes may, in part, account for the excessive secretory response to CT noted in the immature intestine and that enterocyte endocytosis of CT is developmentally regulated. To test this hypothesis, we used specific inhibitors to define endocytic pathways in mature and immature cell lines. We showed that internalization of CT in adult enterocytes is less and occurs via the caveolae/raft-mediated pathway in contrast to an enhanced immature human enterocyte CT uptake that occurs via a clathrin pathway. We also present evidence that this clathrin pathway is developmentally regulated as demonstrated by its response to corticosteroids, a known maturation factor that causes a decreased CT endocytosis by this pathway. PMID- 15790758 TI - Crosstalk between NF-kappaB and beta-catenin pathways in bacterial-colonized intestinal epithelial cells. AB - Salmonella-epithelial cell interactions are known to activate the proinflammatory NF-kappaB signaling pathway and have recently been found to also influence the beta-catenin signaling pathway, an important regulator of epithelial cell proliferation and differentiation. Here, using polarized epithelial cell models, we demonstrate that these same bacteria-mediated effects also direct the molecular crosstalk between the NF-kappaB and beta-catenin signaling pathways. Convergence of these two pathways is a result of the direct interaction between the NF-kappaB p50 subunit and beta-catenin. We show that PhoP(c), the avirulent derivative of a wild-type Salmonella strain, attenuates NF-kappaB activity by stabilizing the association of beta-catenin with NF-kappaB. In cell lines expressing constitutively active beta-catenin, IkappaBalpha protein was indirectly stabilized and NF-kappaB activity was repressed after wild-type Salmonella colonization. Accordingly, constitutively active beta-catenin was found to inhibit the secretion of IL-8. Thus our findings strongly suggest that the crosstalk between the beta-catenin and NF-kappaB signaling pathways is an important regulator of intestinal inflammation. PMID- 15790759 TI - Transcriptional regulation of inflammatory mediators secreted by human colonic circular smooth muscle cells. AB - We investigated the transcriptional regulation of secretion of pro- and anti inflammatory mediators by human colonic circular smooth muscle cells (HCCSMC) in response to tumor necrosis factor (TNF)-alpha. Gene chip array analysis indicated that HCCSMC express a specific panel of 11 cytokines, chemokines, and cell adhesion molecules in a time-dependent manner in response to TNF-alpha. The chip array data were supported by quantitative analysis of mRNA and protein expressions of interleukin (IL)-6, IL-8, intercellular adhesion molecule (ICAM)-1 and IL-11. The proinflammatory mediators were expressed early, whereas the anti inflammatory cytokine IL-11 was expressed late after TNF-alpha treatment. The expression of ICAM-1 on HCCSMC increased lymphocyte adhesion to these cells, which was blocked by pretreatment with antibody to ICAM-1. TNF-alpha acted on both R(1) and R(2) receptors to induce the expression of ICAM-1. Pretreatment of HCCSMC with antisense oligonucleotides to p65 nuclear factor-kappaB (NF-kappaB) blocked the expression of ICAM-1, whereas pretreatment with antisense oligonucleotides to p50 NF-kappaB had little effect. The overexpression of p65 NF kappaB enhanced the constitutive expression of ICAM-1, and TNF-alpha treatment had no further effect. The delayed expression of endogenous IL-11 limited the expression of ICAM-1, and pretreatment of HCCSMC with antisense oligonucleotides to IL-11 enhanced it. We conclude that TNF-alpha induces gene expression in HCCSMC for programmed synthesis and release of pro- and anti-inflammatory mediators. PMID- 15790760 TI - Lipopolysaccharide-induced changes in mesenteric afferent sensitivity of rat jejunum in vitro: role of prostaglandins. AB - Bacterial translocation across the intestinal mucosal barrier leads to a macrophage-mediated inflammatory response, visceral hyperalgesia, and ileus. Our aim was to examine how mediators released into mesenteric lymph following LPS treatment influence intestinal afferent sensitivity and the role played by prostanoids in any sensitization. Intestinal lymph was collected from awake rats following treatment with either saline or LPS (5 mg/kg ip). Extracellular multiunit afferent recordings were made from paravascular mesenteric nerve bundles supplying the rat jejunum in vitro following arterial administration of control lymph, LPS lymph, and LPS. Mesenteric afferent discharge increased significantly after LPS lymph compared with control lymph. Peak discharge occurred within 2 min and remained elevated for 5 to 8 min. This response was attenuated by pretreatment with naproxen (10 microM), and restored upon addition of prostaglandin E(2) (5 microM) in the presence of naproxen, but AH6809 (5 microM), an EP(1)/EP(2) receptor(s) antagonist, failed to decrease the magnitude of LPS lymph-induced response. LPS itself also stimulated mesenteric afferent discharge but was unaffected by naproxen. TNF-alpha was significantly increased in LPS lymph compared with control lymph (1,583 +/- 197 vs. 169 +/- 38 pg/ml, P < 0.01) but exogenous TNF-alpha failed to evoke any afferent nerve discharge. We concluded that inflammatory mediators released from the gut into mesenteric lymph during endotoxemia have a profound effect on afferent discharge. These mediators influence afferent firing via the release of local prostaglandins. PMID- 15790761 TI - Reduced intestinal fat absorptive capacity but enhanced susceptibility to diet induced fatty liver in mice heterozygous for ApoB38.9 truncation. AB - Fatty liver is prevalent in apolipoprotein B (apoB)-defective familial hypobetalipoproteinemia (FHBL). Similar to humans, mouse models of FHBL produced by gene targeting (apob(+/38.9)) manifest low plasma cholesterol and increased hepatic triglycerides (TG) even on a chow diet due to impaired hepatic VLDL-TG secretive capacity. Because apoB truncations shorter than apoB48 are expressed in the intestine, we examined whether FHBL mice may have limited capacity for intestinal dietary TG absorption. In addition, we investigated whether FHBL mice are more susceptible to diet-induced hepatic TG accumulation. Fat absorption capacity was impaired in apoB38.9 mice in a gene dose-dependent manner. Relative fractional fat absorption coefficients for apob(+/+), apob(+/38.9), and apob(38.9/38.9) were 1.00, 0.96, and 0.71, respectively. To raise hepatic TG, we fed high-fat (HF) and low-fat (LF) pellets. Hepatic TG level was observed in rank order: HF > LF > chow. On both LF and HF, liver TG level was higher in the apob(+/38.9) than in apob(+/+). Hepatic TG secretion remained impaired in the apob(+/38.9) on the HF diet. Thus the FHBL mice are more susceptible to diet induced fatty liver despite relatively reduced intestinal TG absorption capacity on a HF diet. PMID- 15790762 TI - Effect of mibefradil on sodium and calcium currents. AB - Interstitial cells of Cajal (ICC) generate the electrical slow wave. The ionic conductances that contribute to the slow wave appear to vary among species. In humans, a tetrodotoxin-resistant Na+ current (Na(V)1.5) encoded by SCN5A contributes to the rising phase of the slow wave, whereas T-type Ca2+ currents have been reported from cultured mouse intestine ICC and also from canine colonic ICC. Mibefradil has a higher affinity for T-type over L-type Ca2+ channels, and the drug has been used in the gastrointestinal tract to identify T-type currents. However, the selectivity of mibefradil for T-type Ca2+ channels over ICC and smooth muscle Na+ channels has not been clearly demonstrated. The aim of this study was to determine the effect of mibefradil on T-type and L-type Ca2+ and Na+ currents. Whole cell currents were recorded from HEK-293 cells coexpressing green fluorescent protein with either the rat brain T-type Ca2+ channel alpha(1)3.3b + beta(2), the human intestinal L-type Ca2+ channel subunits alpha(1C) + beta(2), or Na(V)1.5. Mibefradil significantly reduced expressed T-type Ca2+ current at concentrations > or = 0.1 microM (IC(50) = 0.29 microM), L-type Ca2+ current at > 1 microM (IC(50) = 2.7 microM), and Na+ current at > or = 0.3 microM (IC(50) = 0.98 microM). In conclusion, mibefradil inhibits the human intestinal tetrodotoxin-resistant Na+ channel at submicromolar concentrations. Caution must be used in the interpretation of the effects of mibefradil when several ion channel classes are coexpressed. PMID- 15790763 TI - Drebrin E2 is differentially expressed and phosphorylated in parietal cells in the gastric mucosa. AB - Developmentally regulated brain proteins (drebrins) are highly expressed in brain where they may regulate actin filament formation in dendritic spines. Recently, the drebrin E2 isoform was detected in certain epithelial cell types including the gastric parietal cell. In gastric parietal cells, activation of HCl secretion is correlated with actin filament formation and elongation within intracellular canaliculi, which are the sites of acid secretion. The aim of this study was to define the pattern of drebrin expression in gland units in the intact rabbit oxyntic gastric mucosa and to initiate approaches to define the functions of this protein in parietal cells. Drebrin E2 expression was limited entirely or almost entirely to parietal cells and depended upon the localization of parietal cells along the gland axis. Rabbit drebrin E2 was cloned and found to share 86% identity with human drebrin 1a and to possess a number of cross-species conserved protein-protein interaction and phosphorylation consensus sites. Two-dimensional Western blot and phosphoaffinity column analyses confirmed that drebrin is phosphorylated in parietal cells, and several candidate phosphorylation sites were identified by mass spectrometry. Overexpression of epitope-tagged drebrin E2 led to the formation of microspikes and F-actin-rich ring-like structures in cultured parietal cells and suppressed cAMP-dependent acid secretory responses. In Madin-Darby canine kidney cells, coexpression of epitope-tagged drebrin and the Rho family GTPase Cdc42, which induces filopodial extension, produced an additive increase in the length of microspike projections. Coexpression of dominant negative Cdc42 with drebrin E2 did not prevent drebrin-induced microspike formation. These findings suggest that 1) drebrin can induce the formation of F-actin-rich membrane projections by Cdc42-dependent and independent mechanisms; and that 2) drebrin plays an active role in directing the secretagogue-dependent formation of F-actin-rich filaments on the parietal cell canalicular membrane. Finally, the differential distribution of drebrin in parietal cells along the gland axis suggests that drebrin E2 may be an important marker of parietal cell differentiation and functionality. PMID- 15790765 TI - Identification of developmental pluripotency associated 5 expression in human pluripotent stem cells. AB - Pluripotent embryonic germ cells (EGCs) can be derived from the culture of primordial germ cells (PGCs). However, there are no reports of gonocytes, following the stage of PGC development, becoming stem cell lines. To analyze the gene expression differences between PGCs and gonocytes, we performed cDNA subtractive hybridization with mouse gonads containing either of the two cell populations. We confirmed that developmental pluripotency associated 5 (Dppa5), originally found in mouse embryonic stem cells (ESCs) and mouse embryonic carcinoma cells (ECCs), was strongly expressed in mouse PGCs and the expression was rapidly downregulated during germ cell development. A human sequence homologous to Dppa5 was identified by bioinformatics approaches. Interestingly, human Dppa5 was expressed only in human PGCs, human EGCs, and human ESCs and was not detected in human ECCs. Its expression was downregulated during induced differentiation of human ESCs. These findings confirmed that Dppa5 is specifically and differentially expressed in human cells that have pluripotency. The results strongly suggest that Dppa5 may have an important role in stemness in human ESCs and EGCs and also can be used as a marker of pluripotent stem cells. Human pluripotent stem cells may have their own ways to be pluripotent, as opposed to the much uniform mouse stem cells. PMID- 15790766 TI - Portal application of autologous CD133+ bone marrow cells to the liver: a novel concept to support hepatic regeneration. AB - The liver has a large capacity for regeneration after resection. However, below a critical level of future liver remnant volume (FLRV), partial hepatectomy is accompanied by a significant increase of postoperative liver failure. There is accumulating evidence for the contribution of bone marrow stem cells (BMSCs) to participate in liver regeneration. Here we report on three patients subjected to intraportal administration of autologous CD133(+) BMSCs subsequent to portal venous embolization of right liver segments, used to expand left lateral hepatic segments as FLRV. Computerized tomography scan volumetry revealed 2.5-fold increased mean proliferation rates of left lateral segments compared with a group of three consecutive patients treated without application of BMSCs. This early experience with portovenous application of CD133(+) BMSCs could suggest that this novel therapeutic approach bears the potential of enhancing and accelerating hepatic regeneration in a clinical setting. PMID- 15790767 TI - Cripto as a target for improving embryonic stem cell-based therapy in Parkinson's disease. AB - Embryonic stem (ES) cells have been suggested as candidate therapeutic tools for cell replacement therapy in neurodegenerative disorders. However, limitations for the use of these cells lie in our restricted knowledge of the molecular mechanisms involved in their specialized differentiation and in the risk of tumor formation. Recent findings suggest that the EGF-CFC protein Cripto is a key player in the signaling pathways controlling neural induction in ES cells. Here we show that in vitro differentiation of Cripto(-/-) ES cells results in increased dopaminergic differentiation and that, upon transplantation into Parkinsonian rats, they result in behavioral and anatomical recovery with no tumor formation. The use of knockout ES cells that can generate dopamine cells while eliminating tumor risk holds enormous potential for cell replacement therapy in Parkinson's disease. PMID- 15790768 TI - Magnetic cell sorting purification of differentiated embryonic stem cells stably expressing truncated human CD4 as surface marker. AB - Embryonic stem (ES) cells offer great potential in regenerative medicine and tissue engineering. Clinical applications are still hampered by the lack of protocols for gentle, high-yield isolation of specific cell types for transplantation expressing no immunogenic markers. We describe labeling of stably transfected ES cells expressing a human CD4 molecule lacking its intracellular domain (DeltaCD4) under control of the phosphoglycerate kinase promoter for magnetic cell sorting (MACS). To track the labeled ES cells, we fused DeltaCD4 to an intracellular enhanced green fluorescent protein domain (DeltaCD4EGFP). We showed functionality of the membrane-bound fluorescent fusion protein and its suitability for MACS leading to purities greater than 97%. Likewise, expression of DeltaCD4 yielded up to 98.5% positive cells independently of their differentiation state. Purities were not limited by the initial percentage of DeltaCD4(+) cells, ranging from 0.6%-16%. The viability of MACS-selected cells was demonstrated by reaggregation and de novo formation of embryoid bodies developing all three germ layers. Thus, expression of DeltaCD4 in differentiated ES cells may enable rapid, high-yield purification of a desired cell type for tissue engineering and transplantation studies. PMID- 15790769 TI - High-resolution analysis of the subtelomeric regions of human embryonic stem cells. AB - The use of human embryonic stem cells (hESCs) in most applications is dependent on their undifferentiated proliferation in vitro. Recent studies have illustrated the possibility that chromosomal changes may occur in hESCs during in vitro propagation of these cells. However, no studies so far have screened for chromosomal abnormalities in hESCs using high-resolution techniques that can detect alterations on a few base-pair levels. We have used the recently developed multiplex ligation-dependent probe amplification procedure to analyze the possible occurrence of deletions or duplications in the subtelomeric regions of hESCs in early and late passages. In this study we show that no subtelomeric anomalies were detected in any of the nine hESC lines investigated, supporting the conclusion that hESCs, under appropriate conditions, maintain genomic stability during in vitro propagation. PMID- 15790770 TI - Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers. AB - To date, all human embryonic stem cells (hESCs) available for research require unidentified soluble factors secreted from feeder layers to maintain the undifferentiated state and pluripotency. Activation of STAT3 by leukemia inhibitory factor is required to maintain "stemness" in mouse embryonic stem cells, but not in hESCs, suggesting the existence of alternate signaling pathways for self-renewal and pluripotency in human cells. Here we show that activin A is secreted by mouse embryonic feeder layers (mEFs) and that culture medium enriched with activin A is capable of maintaining hESCs in the undifferentiated state for >20 passages without the need for feeder layers, conditioned medium from mEFs, or STAT3 activation. hESCs retained both normal karyotype and markers of undifferentiated cells, including Oct-4, nanog, and TRA-1-60 and remained pluripotent, as shown by the in vivo formation of teratomas. PMID- 15790771 TI - The kinetic status of hematopoietic stem cell subpopulations underlies a differential expression of genes involved in self-renewal, commitment, and engraftment. AB - The gene expression profile of CD34(-) hematopoietic stem cells (HSCs) and the correlations with their biological properties are still poorly understood. To address this issue, we used the DNA microarray technology to compare the expression profiles of different peripheral blood hemopoietic stem/progenitor cell subsets, lineage-negative (Lin(-)) CD34(-), Lin(-)CD34(+), and Lin(+)CD34(+) cells. The analysis of gene categories differentially expressed shows that the expression of CD34 is associated with cell cycle entry and metabolic activation, such as DNA, RNA, and protein synthesis. Moreover, the significant upregulation in CD34(-) cells of pathways inhibiting HSC proliferation induces a strong differential expression of cyclins, cyclin-dependent kinases (CDKs), CDK inhibitors, and growth-arrest genes. According to the expression of their receptors and transducers, interleukin (IL)-10 and IL-17 showed an inhibitory effect on the clonogenic activity of CD34(-) cells. Conversely, CD34(+) cells were sensitive to the mitogenic stimulus of thrombopoietin. Furthermore, CD34(-) cells express preferentially genes related to neural, epithelial, and muscle differentiation. The analysis of transcription factor expression shows that the CD34 induction results in the upregulation of genes related to self-renewal and lineage commitment. The preferential expression in CD34(+) cells of genes supporting the HSC mobilization and homing to the bone marrow, such as chemokine receptors and integrins, gives the molecular basis for the higher engraftment capacity of CD34(+) cells. Thus, the different kinetic status of CD34(-) and CD34(+) cells, detailed by molecular and functional analysis, significantly influences their biological behavior. PMID- 15790772 TI - CD34+ corneal stromal cells are bone marrow-derived and express hemopoietic stem cell markers. AB - Previous studies have suggested that corneal stromal keratocytes express the CD34 antigen. We wished to investigate CD34 antigen expression in normal mouse cornea using dual- and triple-staining techniques. Whole-mount preparations of mouse and rat corneas were examined with confocal microscopy using single, dual, or triple immunostaining to study their morphology, phenotype, and distribution. Single cell suspensions from normal mouse corneas were also prepared and analyzed by flow cytometry. After short-term culture of corneal stromal explants, nonadherent cells were harvested and cytospins were prepared and stained for different markers. Combined staining for F-actin and leukocyte differentiation markers clearly showed that the corneal stroma contains a population of CD45(+) resident bone marrow-derived cells, whereas most cells were CD45-F-actin(+) keratocytes. A significant proportion (two thirds) of CD45(+) cells in the normal corneal stroma expressed CD34(+), whereas no CD45(-) cells (i.e., keratocytes) coexpressed CD34. In addition, CD34(+) cells were CD11c(-) and CD11b(+). Fewer than 10% of the CD34(+) cells also coexpressed Sca-1(+), but no CD34(+) cells coexpressed major histocompatibility complex (MHC) class II(+). In contrast, the remaining population of CD45(+)CD34(-) cells in the corneal stroma expressed CD11b, MHC class II(+) but not CD11c and were found mostly in the anterior and peripheral part of stroma. These cells are in intimate contact with corneal keratocytes, which stained only for F-actin and were negative for all leukocyte markers. Very few CD45(+) cells expressed the B220 marker, suggesting a plasmacytoid dendritic cell phenotype. Flow cytometry analyses confirmed the morphometric data showing that 68% of CD45(+) cells coexpress CD34 and CD11b, whereas 22% are CD11b(+)CD34( ).We conclude that the normal mouse cornea contains two populations of bone marrow-derived leukocytes, both of which are distinct from stromal keratocytes. The larger population resembles CD34(+) hemopoietic stem cells, whereas the smaller population are CD34(-)CD11b(+) MHC class II(+) macrophages. A very small percentage comprises plasmacytoid dendritic cells. PMID- 15790773 TI - Overexpression of telomerase confers growth advantage, stress resistance, and enhanced differentiation of ESCs toward the hematopoietic lineage. AB - Embryonic stem cells (ESCs) are capable of extended self-renewal and maintenance of pluripotency even after many population doublings. This is supported by high levels of telomerase activity and enhanced antioxidant protection in ESCs, both of which are downregulated during differentiation. To examine the role of telomerase for ESC self-renewal and differentiation, we overexpressed the reverse transcriptase subunit (Tert) of murine telomerase in ESCs. Increased telomerase activity enhances the self-renewal ability of the Tert-overexpressing ESCs, improves their resistance to apoptosis, and increases their proliferation. The differentiated progeny of wild-type ESCs express little Tert and show shortening of telomeric overhangs. In contrast, the progeny of Tert-overexpressing ESCs maintain high telomerase activity, as well as the length of G-rich overhangs. In addition, these cells accumulate lower concentrations of peroxides than wild-type cells, implying greater resistance to oxidative stress. Finally, differentiation toward hematopoietic lineages is more efficient as a result of the continued expression of Tert. Microarray analysis revealed that overexpression of Tert altered expression of a variety of genes required for extended self-renewal and lifespan. Our results suggest that telomerase functions as a "survival enzyme" in ESCs and its differentiated progeny by protecting the telomere cap and by influencing the expression patterns of stress response and defense genes. This results in improved proliferation of ESCs and more efficient differentiation, and these results might have profound consequences for stem cell-replacement therapies. PMID- 15790774 TI - JAK2/STAT3 directs cardiomyogenesis within murine embryonic stem cells in vitro. AB - The heart is the first organ to form during development; however, little is known about the mechanisms that control the initial stages of cardiac differentiation. To investigate this process, we used a protein kinase expression screen, in which nonbeating embryonic stem (ES) cells were compared with beating ES cell-derived cardiomyocytes. We found that JAK2 experienced a 70% increase in protein levels within beating areas. Inhibition of JAK2 pharmacologically or by using dominant/negative JAK2 both resulted in diminished beating within embryoid bodies (EBs), whereas gain of function analysis using dominant/positive JAK2 resulted in a significant induction of beating. More important, inhibition of STAT3, a specific target of JAK2, by dominant/negative STAT3 resulted in the virtual complete loss of beating areas. Reverse transcription-polymerase chain reaction and Western analysis of STAT3-inhibited EBs resulted in lack of expression of several cardiac-specific genes, many of which contain within their promoter STAT3 DNA-binding regions. Taken together, the data reveal that the JAK2/STAT3 pathway is essential for initial stages of cardiomyogenesis. PMID- 15790775 TI - Derivation of human embryonic stem cell lines in serum replacement medium using postnatal human fibroblasts as feeder cells. AB - Derivation and culture of human embryonic stem cells (hESCs) without animal derived material would be optimal for cell transplantation. We derived two new hES (HS293 and HS306) and 10 early cell lines using serum replacement (SR) medium instead of conventional fetal calf serum and human foreskin fibroblasts as feeder cells. Line HS293 has been in continuous culture, with a passage time of 5-8 days, since October 2003 and is at passage level 56. Line HS306 has been cultured since February 2004, now at passage 41. The lines express markers of pluripotent hESCs (Oct-4, SSEA-4, TRA-1-60, TRA-1-81, GCTM-2, and alkaline phosphatase). The pluripotency has been shown in embryoid bodies in vitro, and the pluripotency of line 293 has also been shown in vivo by teratoma formation in severe combined immunodeficiency/beige mice. The karyotype of HS293 is 46,XY, and that of HS306 is 46,XX. Ten more early lines have been derived under similar conditions since September 2004. We conclude that hESC lines can be successfully derived using SR medium and postnatal human fibroblasts as feeder cells. This is a step toward xeno-free conditions and facilitates the use of these cells in transplantation. PMID- 15790776 TI - Notch/Delta4 interaction in human embryonic liver CD34+ CD38- cells: positive influence on BFU-E production and LTC-IC potential maintenance. AB - We investigated whether Notch signaling pathways have a role in human developmental hematopoiesis. In situ histochemistry analysis revealed that Notch1, 2, and 4 and Notch ligand (Delta1-4, and Jagged1) proteins were not expressed in the yolk sac blood islands, the para-aortic splanchnopleure, the hematopoietic aortic clusters, and at the early stages of embryonic liver hematopoiesis. Notch1-2, and Delta4 were eventually detected in the embryonic liver, from 34 until 38 days postconception. Fluorescence-activated cell sorter analysis showed that first-trimester embryonic liver CD34(+)CD38(low) cells expressed both Notch1 and Notch2. When these cells were cultured on S17 stroma stably expressing Delta4, a 2.6-fold increase in BFU-E number was observed at day 7, as compared with cultures with control stroma, and this effect was maintained for 2 weeks. Importantly, exposure of these cells to Delta4 under these conditions maintained the original frequency and quality of long-term culture initiating cells (LTC-ICs), while control cultures quickly resulted in the extinction of this LTC-IC potential. Furthermore, short-term exposure of embryonic liver adherent cells to erythropoietin resulted in a dose-dependent increase in Delta4 expression, almost doubling the expression observed with untreated stroma. This suggests that Delta4 has a role in the regulation of hematopoiesis after a hypoxic stress in the fetus. PMID- 15790777 TI - Cycling G1 CD34+/CD38+ cells potentiate the motility and engraftment of quiescent G0 CD34+/CD38-/low severe combined immunodeficiency repopulating cells. AB - The mechanism of human stem cell expansion ex vivo is not fully understood. Furthermore, little is known about the mechanisms of human stem cell homing/repopulation and the role that differentiating progenitor cells may play in these processes. We report that 2- to 3-day in vitro cytokine stimulation of human cord blood CD34(+)-enriched cells induces the production of short-term repopulating, cycling G1 CD34(+)/CD38(+) cells with increased matrix metalloproteinase (MMP)-9 secretion as well as increased migration capacity to the chemokine stromal cell-derived factor-1 (SDF-1) and homing to the bone marrow of irradiated nonobese diabetic severe/combined immunodeficiency (NOD/SCID) mice. These cycling G1 cells enhance SDF-1-mediated in vitro migration and in vivo homing of quiescent G0 CD34(+) cells, which is partially abrogated after inhibition of MMP-2/-9 activity. Moreover, the engraftment potential of quiescent G0 SCID repopulating cells (SRCs) is also increased by the cycling G1 CD34(+)/CD38(+) cells. This effect is significantly abrogated after incubation of cycling G1 cells with a neutralizing anti-CXCR4 antibody. Our data suggest synergistic interactions between accessory cycling G1 CD34(+)/CD38(+) committed progenitor cells and quiescent, primitive G0 CD34(+)/CD38(-/low) SRC/stem cells, the former increasing the motility and engraftment potential of the latter, partly via secretion of MMP-9. PMID- 15790778 TI - Obese diabetic mouse environment differentially affects primitive and monocytic endothelial cell progenitors. AB - Two classes of adult bone marrow-derived endothelial cell (EC) progenitors have been described, primitive hematopoietic stem cell-related cells and monocytic cells. Both differentiate into ECs and promote vascular growth in vivo but have distinct characteristics. Despite the association of obesity and type 2 diabetes with cardiovascular disease, their effects on primitive EC progenitors (prECPs) have not been examined, and the limited data on monocytic EC progenitors are conflicting. We investigated functional parameters of primitive and monocytic EC progenitors from obese diabetic (Lepr(db)) mice. The viability, proliferation, and differentiation of EC progenitors were unaffected in Lepr(db) cell cultures under basal condition. However, Lepr(db)-derived prECPs, but not monocytic EC progenitors, were less able to cope with hypoxia and oxidative stress, conditions likely present when EC progenitors are most needed. Intrinsic prECP dysfunction was also apparent in vivo. Whereas injection of nondiabetic prECPs promoted vascularization of skin wounds, Lepr(db)-derived progenitors inhibited it in nondiabetic mice. Additionally, although treatment with Lepr(db)-derived prECPs did not significantly reduce blood flow restoration to ischemic limbs, it resulted in increased tissue necrosis and autoamputation. Thus, type 2 diabetes coupled with obesity seems to induce intrinsic EC progenitor dysfunction that is exacerbated by stress. prECPs are more affected than monocytic progenitors, exhibiting a reduced ability to survive or proliferate. The proangiogenic phenotype of prECPs also seems to convert to an antiangiogenic phenotype in obese diabetic mice. These data suggest that therapies involving prECPs or stem-like cells in diabetic patients may be inadvisable at this time. PMID- 15790779 TI - Differential gene expression profiling of human umbilical cord blood-derived mesenchymal stem cells by DNA microarray. AB - Mesenchymal stem cells (MSCs) retain both self-renewal and multilineage differentiation capabilities. Despite wide therapeutic potential, many aspects of human MSCs, particularly the molecular parameters to define the stemness, remain largely unknown. Using high-density oligonucleotide micro-arrays, we obtained the differential gene expression profile between a fraction of mononuclear cells of human umbilical cord blood (UCB) and its MSC subpopulation. Of particular interest was a subset of 47 genes preferentially expressed at 50-fold or higher in MSCs, which could be regarded as a molecular foundation of human MSCs. This subset contains numerous genes encoding collagens, other extracellular matrix or related proteins, cytokines or growth factors, and cytoskeleton-associated proteins but very few genes for membrane and nuclear proteins. In addition, a direct comparison of this microarray-generated transcriptome with the published serial analysis of gene expression data suggests that a molecular context of UCB derived MSCs is more or less similar to that of bone marrow-derived cells. Altogether, our results will provide a basis for studies on molecular mechanisms controlling core properties of human MSCs. PMID- 15790780 TI - Human bone marrow mesenchymal stem cells can express insulin and key transcription factors of the endocrine pancreas developmental pathway upon genetic and/or microenvironmental manipulation in vitro. AB - Multipotential stem cells can be selected from the bone marrow by plastic adhesion, expanded, and cultured. They are able to differentiate not only into multiple cell types, including cartilage, bone, adipose and fibrous tissues, and myelosupportive stroma, but also into mesodermal (endothelium), neuroectodermal, or endodermal (hepatocytes) lineages. Our goal was to characterize the multipotential capacities of human mesenchymal stem cells (hMSCs) and to evaluate their ability to differentiate into insulin-secreting cells in vitro. hMSCs were obtained from healthy donors, selected by plastic adhesion, and phenotyped by fluorescence-activated cell sorter and reverse transcription-polymerase chain reaction analysis before and after infection with adenoviruses coding for mouse IPF1, HLXB9, and FOXA2 transcription factors involved early in the endocrine developmental pathway. We found that native hMSCs have a pluripotent phenotype (OCT4 expression and high telomere length) and constitutively express NKX6-1 at a low level but lack all other transcription factors implicated in beta-cell differentiation. In all hMSCs, we detected mRNA of cytokeratin 18 and 19, epithelial markers present in pancreatic ductal cells, whereas proconvertase 1/3 mRNA expression was detected only in some hMSCs. Ectopic expression of IPF1, HLXB9, and FOXA2 with or without islet coculture or islet-conditioned medium results in insulin gene expression. In conclusion, our results demonstrated that in vitro human bone marrow stem cells are able to differentiate into insulin expressing cells by a mechanism involving several transcription factors of the beta-cell developmental pathway when cultured in an appropriate microenvironment. PMID- 15790781 TI - Hematologic differences between African-Americans and whites: the roles of iron deficiency and alpha-thalassemia on hemoglobin levels and mean corpuscular volume. AB - The average results of some laboratory measurements, including the hemoglobin, mean corpuscular volume (MCV), serum transferrin saturation (TS), serum ferritin, and white blood cell count of African-Americans differ from those of whites. Anonymized samples and laboratory data from 1491 African-American and 31 005 white subjects, approximately equally divided between men and women, were analyzed. The hematocrit, hemoglobin, MCV, TS, and white blood cell counts of African-Americans were lower than those of whites; serum ferritin levels were higher. When iron-deficient patients were eliminated from consideration the differences in hematocrit, hemoglobin, and MCV among women were slightly less. The -3.7-kilobase alpha-thalassemia deletion accounted for about one third of the difference in the hemoglobin levels of African-Americans and whites and neither sickle trait nor elevated creatinine levels had an effect. Among all subjects, 19.8% of African-American women would have been classified as "anemic" compared with 5.3% of whites. Among men, the figures were 17.7% and 7.6%. Without iron deficient or thalassemic subjects, the difference had narrowed to 6.1% and 2.77% and to 4.29% and 3.6%, respectively. Physicians need to take into account that the same reference standards for hemoglobin, hematocrit, MCV, and TS and the white blood cell count do not apply to all ethnic groups. PMID- 15790782 TI - Cytochrome P450 2C9 (CYP2C9) and vitamin K epoxide reductase (VKORC1) genotypes as determinants of acenocoumarol sensitivity. AB - The aim of the study is to explore the contribution of genetic factors related either to drug metabolism (cytochrome P450 2C9) or to drug target (vitamin K epoxide reductase) to variability in the response to acenocoumarol among 222 healthy volunteers after a single oral dose. Associations between a pharmacodynamic index (reduction in factor VII activity and international normalized ratio [INR] change) and several genetic polymorphisms (VKORC1: 4931T>C, -4451C>A, -2659G>C, -1877A>G, -1639G>A, 497C>G, 1173C>T, and CYP2C9*3) were investigated using haplotype and univariate analyses. VKORC1 haplotypes were associated with the pharmacologic response, and this association can be explained only by the effect of the -1639G>A polymorphism (or alternatively by 1173C>T, which is in complete association with it). Indeed, it explains about one third of the variability of the pharmacologic response (37% of factor VII decrease and 30% of INR change). Moreover, the previously observed effect of the CYP2C9*3 allele is independent of the VKORC1 gene effect. These 2 polymorphisms account for up to 50% of the interindividual variability. The simple genotyping of 2 single nucleotide polymorphisms (SNPs), VKORC1 -1639G>A or 1173C>T and the CYP2C9*3 polymorphisms, could thus predict a high risk of overdose before initiation of anticoagulation with acenocoumarol, and provide a safer and more individualized anticoagulant therapy. PMID- 15790783 TI - Progressive neurologic dysfunctions 20 years after allogeneic bone marrow transplantation for Chediak-Higashi syndrome. AB - Three patients with Chediak-Higashi syndrome underwent allogeneic bone marrow transplantation between the ages of 2 years 9 months and 7 years. The outcome was uneventful, with sustained mixed chimerism. No subsequent recurrent infections or hemophagocytic syndrome were observed. At the age of 22 to 24 years, these 3 patients developed a neurologic deficit combining difficulty walking, loss of balance, and tremor. Neurologic evaluation demonstrated cerebellar ataxia and signs of peripheral neuropathy. Moderate axon loss and rarefaction of large myelinated fibers were observed on semithin sections of peripheral nerve. Cerebellar atrophy was detected by cerebral magnetic resonance imaging in 2 patients. We also reviewed the very long-term outcome of the other 11 patients with Chediak-Higashi syndrome who had received bone marrow transplants at our center since 1981. All displayed neurologic deficits or low cognitive abilities. PMID- 15790784 TI - ICAM-1 on exosomes from mature dendritic cells is critical for efficient naive T cell priming. AB - Exosomes are secreted vesicles formed in late endocytic compartments. Immature dendritic cells (DCs) secrete exosomes, which transfer functional major histocompatibility complex (MHC)-peptide complexes to other DCs. Since immature and mature DCs induce different functional T-cell responses (ie, tolerance versus priming), we asked whether DC maturation also influenced the priming abilities of their exosomes. We show that exosomes secreted by lipopolysaccharide (LPS) treated mature DCs are 50- to 100-fold more potent to induce antigen-specific T cell activation in vitro than exosomes from immature DCs. In vitro, exosomes from mature DCs transfer to B lymphocytes the ability to prime naive T cells. In vivo, only mature exosomes trigger effector T-cell responses, leading to fast skin graft rejection. Proteomic and biochemical analyses revealed that mature exosomes are enriched in MHC class II, B7.2, intercellular adhesion molecule 1 (ICAM-1), and bear little milk-fat globule-epidermal growth factor-factor VIII (MFG-E8) as compared with immature exosomes. Functional analysis using DC-derived exosomes from knock-out mice showed that MHC class II and ICAM-1 are required for mature exosomes to prime naive T cells, whereas B7.2 and MFG-E8 are dispensable. Therefore, changes in protein composition and priming abilities of exosomes reflect the maturation signals received by DCs. PMID- 15790785 TI - Ligand-induced conformational change in the T-cell receptor associated with productive immune synapses. AB - Triggering of the T-cell receptor (TCR) can produce very different responses, depending on the nature of the major histocompatibility complex/antigen peptide (MHCp) ligand. The molecular mechanisms that permit such fine discrimination are still unknown. We show here that an epitope in the cytoplasmic tail of the TCR CD3epsilon subunit, recognized by antibody APA1/1, is only detected when the TCR is fully activated. Exposure of the APA1/1 epitope is shown to be fast and independent of tyrosine kinase activity and that it takes place even when T cells are stimulated at 0 degrees C. These results suggest that APA1/1 detects a conformational change in the TCR. APA1/1 staining concentrates in a restricted area of the immunologic synapse. Most important, we show that full agonist, but not partial agonist, peptides induce exposure of the APA1/1 epitope, indicating a correlation between the induction of the conformational change in the TCR and full T-cell activation. Finally, the conformational change is shown to occur in T cells that are being stimulated by antigen in vivo. Therefore, these results demonstrate that the TCR undergoes a conformational change on MHCp binding in vitro and in vivo, and they establish a molecular correlate for productive TCR engagement. PMID- 15790786 TI - Activation mutations of human c-KIT resistant to imatinib mesylate are sensitive to the tyrosine kinase inhibitor PKC412. AB - Constitutively activated forms of the transmembrane receptor tyrosine kinase c KIT have been associated with systemic mast cell disease, acute myeloid leukemia, and gastrointestinal stromal tumors. Reports of the resistance of the kinase domain mutation D816V to the adenosine triphosphate (ATP)-competitive kinase inhibitor imatinib mesylate prompted us to characterize 14 c-KIT mutations reported in association with human hematologic malignancies for transforming activity in the murine hematopoietic cell line Ba/F3 and for sensitivity to the tyrosine kinase inhibitor PKC412. Ten of 14 c-KIT mutations conferred interleukin 3 (IL-3)-independent growth. c-KIT D816Y and D816V transformed cells were sensitive to PKC412 despite resistance to imatinib mesylate. In these cells, PKC412, but not imatinib mesylate, inhibited autophosphorylation of c-KIT and activation of downstream effectors signal transducer and transcriptional activator 5 (Stat5) and Stat3. Variable sensitivities to PKC412 or imatinib mesylate were observed among other mutants. These findings suggest that PKC412 may be a useful therapeutic agent for c-KIT-positive malignancies harboring the imatinib mesylate-resistant D816V or D816Y activation mutations. PMID- 15790788 TI - Azidothymidine inhibits NF-kappaB and induces Epstein-Barr virus gene expression in Burkitt lymphoma. AB - The antiviral compound azidothymidine (AZT), alone or in combination with other agents, induces apoptosis in early-passage, Epstein-Barr virus-positive Burkitt lymphoma (EBV+ BL) lines and has clinical activity in EBV+ BL. We report here a mechanism of AZT's antitumor activity. The nuclei of these cells contain activated nuclear factor-kappaB (NF-kappaB) subunits p50, c-Rel, RelB, and p52, but not p65. Treatment of primary EBV+ BL lines with AZT inhibited NF-kappaB within 1 to 2 hours. This was followed by up-regulation of EBV gene expression including viral thymidine kinase (vTK) and apoptosis. Subclones of EBV+ BL cells that demonstrated activated p65 were resistant to AZT. In EBV+ BLs, AZT but not ganciclovir (GCV) was highly phosphorylated to its monophosphate form (AZT-MP). Phosphorylation, as well as apoptosis, was markedly enhanced in the presence of hydroxyurea. AZT inhibits NF-kappaB and up-regulates EBV gene expression in primary EBV+ BLs. AZT with hydroxyurea may represent an inexpensive, targeted regimen for endemic BL. PMID- 15790789 TI - Generation of tumor-specific, HLA class I-restricted human Th1 and Tc1 cells by cell engineering with tumor peptide-specific T-cell receptor genes. AB - Tumor antigen-specific CD4+ and CD8+ T lymphocytes, especially interferon-gamma (IFN-gamma)-producing type-1 helper T (Th1) and type-1 cytotoxic T (Tc1) cells, play a crucial role in tumor eradication. Adoptive transfer using tumor-specific Th1 and Tc1 cells is a promising therapeutic strategy for tumor immunotherapy. However, its clinical application has been hampered because of difficulties in generating tumor-specific Th1 cells from patients with tumors. To overcome this problem, we have developed an efficient method to prepare tumor-specific Th1 and Tc1 cells. T-cell receptor (TCR) alpha and beta genes obtained from an HLA-A24 restricted, Wilms tumor 1 (WT1) peptide-specific Tc clone were lentivirally transduced to polyclonally activated Th1 and Tc1 cells. As expected, TCR gene modified Tc1 cells showed cytotoxicity and IFN-gamma production in response to peptide-loaded lymphoblastoid cell lines, WT1 gene-transduced cells, and freshly isolated leukemia cells expressing both WT1 and HLA-A24. Surprisingly, we further demonstrated that Th1 cells transduced with HLA-class I-restricted TCR genes also showed both cytotoxicity and cytokine production in an HLA-A24-restricted manner. In contrast to gene-modified Tc1 cells, Th1 cells produced high amounts of interleukin-2 (IL-2) in addition to IFN-gamma, which is beneficial for induction of antitumor cellular immunity. Thus, TCR gene-modified HLA-class I-restricted Th1 and Tc1 cells are a powerful strategy for the application to adoptive immunotherapy of human cancer. PMID- 15790787 TI - Differential regulation of the p70 S6 kinase pathway by interferon alpha (IFNalpha) and imatinib mesylate (STI571) in chronic myelogenous leukemia cells. AB - The precise mechanisms by which imatinib mesylate (STI571) and interferon alpha (IFNalpha) exhibit antileukemic effects are not known. We examined the effects of IFNs or imatinib mesylate on signaling pathways regulating initiation of mRNA translation in BCR-ABL-expressing cells. Treatment of IFN-sensitive KT-1 cells with IFNalpha resulted in phosphorylation/activation of mammalian target of rapamycin (mTOR) and downstream activation of p70 S6 kinase. The IFN-activated p70 S6 kinase was found to regulate phosphorylation of S6 ribosomal protein, which regulates translation of mRNAs with oligopyrimidine tracts in the 5' untranslated region. In addition, IFNalpha treatment resulted in an mTOR- and/or phosphatidyl-inositol 3'(PI 3') kinase-dependent phosphorylation of 4E-BP1 repressor of mRNA translation on sites that are required for its deactivation and dissociation from the eukaryotic initiation factor-4E (eIF4E) complex. In contrast to the effects of IFNs, imatinib mesylate suppressed p70 S6 kinase activity, consistent with inhibition of BCR-ABL-mediated activation of the mTOR/p70 S6 kinase pathway. Moreover, the mTOR inhibitor rapamycin enhanced the suppressive effects of imatinib mesylate on primary leukemic granulocyte macrophage-colony-forming unit (CFU-GM) progenitors from patients with chronic myelogenous leukemia (CML). Taken altogether, our data demonstrate that IFNs and imatinib mesylate differentially regulate PI 3' kinase/mTOR-dependent signaling cascades in BCR-ABL-transformed cells, consistent with distinct effects of these agents on pathways regulating mRNA translation. They also support the concept that combined use of imatinib mesylate with mTOR inhibitors may be an appropriate future therapeutic strategy for the treatment of CML. PMID- 15790792 TI - FDA labors over NSAID decisions: panel suggests COX-2 inhibitors stay available. PMID- 15790791 TI - A novel missense mutation in ABCA1 results in altered protein trafficking and reduced phosphatidylserine translocation in a patient with Scott syndrome. AB - Scott syndrome (SS) is a bleeding disorder characterized by a failure to expose phosphatidylserine (PS) to the outer leaflet of the platelet plasma membrane. Because the adenosine triphosphate (ATP)-binding cassette transporter A1 (ABCA1) is implicated in the exofacial translocation of PS, we assessed its role in the pathophysiology of a patient with SS. Substantially reduced levels of ABCA1 mRNA were found in the patient's leukocytes, compared with controls. The SS patient was heterozygous for a novel missense mutation c.6064G>A (ABCA1 R1925Q), absent from unaffected family members and controls. Both mutant and wild-type alleles were reduced in mRNA expression, and no causative mutation for this phenomenon was identified in the ABCA1 gene or its proximal promoter, suggesting a putative second mutation in a trans-acting regulatory gene may also be involved in the disorder in this patient. In vitro expression studies showed impaired trafficking of ABCA1 R1925Q to the plasma membrane. Overexpression of wild-type ABCA1 in SS lymphocytes complemented the Ca2+-dependent PS exposure at the cell surface. These data identify a mutation in ABCA1 that contributes to the defective PS translocation phenotype in our patient with SS. PMID- 15790790 TI - Distinct hematopoietic progenitor compartments are delineated by the expression of aldehyde dehydrogenase and CD34. AB - A broad range of hematopoietic stem cells and progenitors reside within a fraction of umbilical cord blood (UCB) that exhibits low light scatter properties (SSC(lo)) and high expression of aldehyde dehydrogenase (ALDH(br)). Many SSC(lo) ALDH(br) cells coexpress CD34; however, other cells express either ALDH or CD34. To investigate the developmental potential of these cell subsets, purified ALDH(br) CD34+, ALDH(neg) CD34+, and ALDH(br) CD34(neg) UCB cells were characterized within a variety of in vivo and in vitro assays. Primitive progenitors capable of multilineage development were monitored in long- and short term repopulation assays performed on nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice, and in primary and secondary long-term culture assays. These progenitors were highly enriched within the ALDH(br) CD34+ fraction. This cell fraction also enriched short-term myeloid progenitors that were detected in vitro. By comparison, ALDH(neg) CD34+ cells contained few primitive progenitors and had diminished short-term myeloid potential but exhibited enhanced short-term natural killer (NK) cell development in vitro. The ALDH(br) CD34(neg) cells were not efficiently supported by any of the assays used. These studies suggested that in particular the expression of ALDH delineated distinct CD34+ stem cell and progenitor compartments. The differential expression of ALDH may provide a means to explore normal and malignant processes associated with myeloid and lymphoid development. PMID- 15790793 TI - Government answers need for cultural competency resource. PMID- 15790794 TI - Defense and Veterans Affairs move toward patient information sharing. PMID- 15790795 TI - Glaucoma and its treatment: a review. AB - PURPOSE: A review of glaucoma and its treatment is presented. SUMMARY: Glaucoma is a common eye disease that can cause irreversible blindness if left undiagnosed and untreated. Glaucoma is a leading cause of blindness in the United States and other industrialized countries. In most cases, the symptoms of early-stage glaucoma are minimal or nonexistent. There are several different types of glaucoma, and they have been classically divided into the categories of primary or secondary open-angle or angle-closure glaucoma. Secondary forms of glaucoma are caused by various ocular or systemic diseases. Every available treatment to prevent progressive glaucomatous optic neuropathy has potential adverse effects and involves a certain amount of risk and financial expense. Conventional first line treatment of glaucoma usually begins with the use of a topical selective or nonselective beta-blocker or a topical prostaglandin analog. Second-line drugs of choice include alpha-agonists and topical carbonic anhydrase inhibitors. Parasympathomimetic agents, most commonly pilocarpine, are considered third-line treatment options. For patients who do not respond to antiglaucoma medications, laser trabeculoplasty and incisional surgery are further methods that can be used to lower intraocular pressure. The results of clinical trials have reaffirmed the utility of antiglaucoma medications in slowing the progression of the disease. CONCLUSION: Glaucoma is a common eye disease that is usually associated with an elevated intraocular pressure. Treatment options for patients with glaucoma include medications, laser therapy, and incisional surgery. The risks and benefits of each type of treatment must be carefully considered to maximize the treatment's benefits while minimizing adverse effects. PMID- 15790796 TI - Rationale for using serotonergic agents to treat irritable bowel syndrome. AB - PURPOSE: The role of serotonin in gastrointestinal (GI)-tract functioning, the pharmacologic rationale for using serotonergic agents in the treatment of irritable bowel syndrome (IBS), and clinical experience with novel serotonergic agents are described. SUMMARY: IBS is a common multisymptom disorder that is associated with a high socioeconomic burden. The goal of treatment is to provide rapid and sustained global relief of the multiple symptoms of IBS with a single, effective, well-tolerated agent. Traditional treatment options target single symptoms, and many patients are dissatisfied with the level of relief achieved and adverse effects. Research has revealed that serotonin is involved in three major actions in the gut: (1) mediating intestinal motility, (2) mediating intestinal secretion in the GI tract, and (3) modulating perception in the bowels. Serotonin is also a vital link in the brain-gut axis. Alterations in key elements of serotonin signaling have been demonstrated in patients with IBS. Tegaserod, a selective serotonin type 4 (5-HT(4))-receptor partial agonist, is indicated for use in women with IBS whose primary bowel symptom is constipation. Alosetron, a 5-HT(3)-receptor antagonist, is indicated for use in women with severe diarrhea-predominant IBS in whom traditional therapies have failed. The clinical usefulness of several other serotonergic agents for IBS is being investigated. CONCLUSION: The use of serotonergic agents in patients with IBS is based on the critical role that serotonin plays in the maintenance of normal gut function and brain-gut communication. Pharmacologic therapies targeting specific serotonin receptors represent an important step in the management of IBS. PMID- 15790798 TI - Causes of hyperglycemia and hypoglycemia in adult inpatients. AB - PURPOSE: The underlying causes of hyperglycemia and hypoglycemia in adult medical and surgical inpatients were studied. METHODS: Hyperglycemic and hypoglycemic events occurring in adult medical and surgical patients admitted between February and July 2003 to a tertiary care hospital were identified prospectively from automated daily printouts of abnormal blood glucose levels generated by the hospital laboratory. Information on the causes of a random sample of events was ascertained within 24 hours through chart review and provider and patient interviews. Narratives were presented to an expert committee to assess the causes of each event and preventability. RESULTS: Eighteen of 24 hypoglycemic events and 26 of 26 hyperglycemic episodes were considered preventable. Failure to adjust antidiabetic drugs in response to decreases in oral intake and unexpected deviation from normal hospital routine were the most common factors contributing to hypoglycemia. Hyperglycemia was most often associated with an unwillingness of providers to take responsibility for diabetes management and the exclusive use of sliding-scale insulin regimens. CONCLUSION: Hyperglycemia and hypoglycemia in medical and surgical inpatients were mostly related to inadequate prescribing, monitoring, and communication practices. PMID- 15790799 TI - Use of i.v. immune globulin and occurrence of associated acute renal failure and thrombosis. AB - PURPOSE: The use trends of i.v. immune globulin (IGIV) and the frequency of acute renal failure (ARF) and thrombosis at one institution were studied. METHODS: A retrospective chart review on the indications and dosages of IGIV was conducted with inpatients and outpatients at a Veterans Affairs medical center between May 1, 1998, and June 30, 2003. Patients under 18 years of age were excluded from the study. Patient data were obtained through the center's computerized patient record system and included patient demographics, comorbidities, frequency of ARF, concomitant therapies, and IGIV therapy courses. ARF was defined as an increase in serum creatinine of >/=0.5 mg/dL within 10 days from the initiation of IGIV therapy. RESULTS: Forty-six patients were identified as receiving at least one cycle of IGIV therapy between May 1, 1998, and June 30, 2003. The three main indications for IGIV therapy were hypogammaglobulinemia, idiopathic thrombocytopenic purpura (ITP), and chronic inflammatory demyelinating polyneuropathy (CIDP). No new cases of thrombosis were found. Six patients (13%) developed ARF after IGIV administration, but none required dialysis. A greater percentage of patients with ARF were 65 years or older, had chronic renal insufficiency or diabetes mellitus, and used nephrotoxic agents, compared with patients who did not develop ARF. No association was observed between ARF and underlying primary disease. CONCLUSION: A retrospective review of 46 courses of IGIV therapy showed that the therapy was most often used for hematologic, neurologic, and immunologic indications, and the most common diagnoses for which it was used were hypogammaglobulinemia, ITP, and CIDP. ARF developed in 13% of patients, but no case of thrombosis was reported. PMID- 15790800 TI - Patient-assistance programs: assessment of and use by safety-net clinics. AB - PURPOSE: Safety-net clinics' use and assessment of patient-assistance programs (PAPs) were studied. METHODS: A multistate telephone survey was conducted on the basis of issues identified during 10 case-study interviews of safety-net clinics serving primarily uninsured and publicly insured patients. Interviewed were pharmacists and other staff taking primary responsibility for helping patients apply to PAPs. RESULTS: Of 339 survey candidates, 215 provided complete interviews (63% response rate). Ninety-three percent of the completed interviews were with clinics in California, Texas, and Florida. Forty percent of the clinics reported that at least 75% of their patients lacked drug insurance coverage. There was a significant positive relationship between a clinic's likelihood of using PAPs and the percentage of its patients lacking drug coverage. PAPs consumed 12 hours of pharmacist time per month and 99 hours of other staff time per month. Clinics most frequently cited program requirements changing without notice and unrealistic income-documentation rules as potential barriers to PAP use and indicated that consistent eligibility criteria and standardized application procedures were needed. CONCLUSION: A survey of safety-net clinics indicated that PAPs help fill a major gap in health insurance coverage but that consistent eligibility criteria and application procedures are needed. PMID- 15790801 TI - Results of an antimicrobial control program at a university hospital. AB - PURPOSE: The results of the first five years of an ongoing antimicrobial control program are reported. METHODS: In 1998, a multidisciplinary antimicrobial subcommittee of the pharmacy and therapeutics committee of a university hospital was formed and charged with making formulary interventions in an effort to reduce rising antimicrobial resistance rates and drug expenditures. In 1999, a number of measures were implemented for controlling antimicrobial use. Selected antimicrobials with the potential for inappropriate use or whose inappropriate use had been documented were placed in the control of physicians in the infectious diseases (ID) division. Prior approval by an ID physician was required before the pharmacy could dispense these agents. Other key interventions included removal of ceftazidime and cefotaxime from the formulary, restriction of vancomycin and carbapenem use, and replacement of ciprofloxacin with levofloxacin as the sole fluoroquinolone on the formulary. Data regarding antimicrobial use and expenditures between 1998 and 2002 were compared and analyzed. RESULTS: Antimicrobial use was reduced by 80% for third-generation cephalosporins and 15% for vancomycin following the implementation of the new antimicrobial policies. Antimicrobial-resistance patterns for many important gram-negative pathogens, including Pseudomonas aeruginosa, demonstrated a reversal of previous increases. In addition, the rate of methicillin-resistant Staphylococcus aureus decreased by an average of 3% each year from 1999 to 2002. Pharmacy expenditures for all antimicrobials, including antiviral, antifungal, and antibacterial agents, decreased 24.7%, with a cumulative cost saving of 1,401,126 US dollars, without inflation assumptions. CONCLUSION: The implementation of an antimicrobial control program decreased the use of selected antimicrobial agents and resulted in substantial reduction of expenditures for antimicrobials. PMID- 15790802 TI - Patient characteristics associated with outpatient prescriptions for nabumetone and oxaprozin versus celecoxib and rofecoxib. PMID- 15790803 TI - Clinical pharmacy in an inpatient leukemia and bone marrow transplant service. PMID- 15790804 TI - ASHP professional policy recommendations--invitation to comment. PMID- 15790805 TI - A world away. PMID- 15790806 TI - Functional genomic analysis of RNA interference in C. elegans. AB - RNA interference (RNAi) of target genes is triggered by double-stranded RNAs (dsRNAs) processed by conserved nucleases and accessory factors. To identify the genetic components required for RNAi, we performed a genome-wide screen using an engineered RNAi sensor strain of Caenorhabditis elegans. The RNAi screen identified 90 genes. These included Piwi/PAZ proteins, DEAH helicases, RNA binding/processing factors, chromatin-associated factors, DNA recombination proteins, nuclear import/export factors, and 11 known components of the RNAi machinery. We demonstrate that some of these genes are also required for germline and somatic transgene silencing. Moreover, the physical interactions among these potential RNAi factors suggest links to other RNA-dependent gene regulatory pathways. PMID- 15790807 TI - Transcriptional maps of 10 human chromosomes at 5-nucleotide resolution. AB - Sites of transcription of polyadenylated and nonpolyadenylated RNAs for 10 human chromosomes were mapped at 5-base pair resolution in eight cell lines. Unannotated, nonpolyadenylated transcripts comprise the major proportion of the transcriptional output of the human genome. Of all transcribed sequences, 19.4, 43.7, and 36.9% were observed to be polyadenylated, nonpolyadenylated, and bimorphic, respectively. Half of all transcribed sequences are found only in the nucleus and for the most part are unannotated. Overall, the transcribed portions of the human genome are predominantly composed of interlaced networks of both poly A+ and poly A- annotated transcripts and unannotated transcripts of unknown function. This organization has important implications for interpreting genotype phenotype associations, regulation of gene expression, and the definition of a gene. PMID- 15790809 TI - Cancers of the gut and Western ills. PMID- 15790808 TI - ATM activation by DNA double-strand breaks through the Mre11-Rad50-Nbs1 complex. AB - The ataxia-telangiectasia mutated (ATM) kinase signals the presence of DNA double strand breaks in mammalian cells by phosphorylating proteins that initiate cell cycle arrest, apoptosis, and DNA repair. We show that the Mre11-Rad50-Nbs1 (MRN) complex acts as a double-strand break sensor for ATM and recruits ATM to broken DNA molecules. Inactive ATM dimers were activated in vitro with DNA in the presence of MRN, leading to phosphorylation of the downstream cellular targets p53 and Chk2. ATM autophosphorylation was not required for monomerization of ATM by MRN. The unwinding of DNA ends by MRN was essential for ATM stimulation, which is consistent with the central role of single-stranded DNA as an evolutionarily conserved signal for DNA damage. PMID- 15790810 TI - Paleoanthropology. Discoverers charge damage to 'hobbit' specimens. PMID- 15790811 TI - Ethics. Doctors pay a high price for priority. PMID- 15790812 TI - Career transitions. Panel throws lifeline to bio postdocs. PMID- 15790813 TI - Scientific misconduct. Researcher faces prison for fraud in NIH grant applications and papers. PMID- 15790814 TI - Genetics. Talking about a revolution: hidden RNA may fix mutant genes. PMID- 15790815 TI - Paleontology. Tyrannosaurus rex soft tissue raises tantalizing prospects. PMID- 15790816 TI - Astronomy. Alien planets glimmer in the heat. PMID- 15790817 TI - Paleoclimate. Ocean flow amplified, not triggered, climate change. PMID- 15790818 TI - Proteomics. Protein chips map yeast kinase network. PMID- 15790819 TI - Magnetic imaging. Atom-based detector puts new twist on nuclear magnetic resonance. PMID- 15790820 TI - Ecology. Savannah River Lab faces budget ax. PMID- 15790821 TI - Proposition 71. Proposed legislation threatens to slow California stem cell rush. PMID- 15790822 TI - Epidemiology. Mounting evidence indicts fine-particle pollution. PMID- 15790823 TI - Epidemiology. How dirty air hurts the heart. PMID- 15790824 TI - Epidemiology. Regulations spark technology competition. PMID- 15790825 TI - U.S. education research. Can randomized trials answer the question of what works? PMID- 15790826 TI - Astronomy. American astronomers lobby for the next big thing. PMID- 15790827 TI - Infectious diseases. True numbers remain elusive in bird flu outbreak. PMID- 15790828 TI - Abuse of prisoners at Abu Ghraib. PMID- 15790829 TI - Reinventing the wheel in ecology research? PMID- 15790830 TI - A central repository for published plasmids. PMID- 15790831 TI - Ethics: a weapon to counter bioterrorism. PMID- 15790832 TI - Physics. Bose-Einstein condensates interfere and survive. PMID- 15790833 TI - Cell biology. Whither model organism research? PMID- 15790834 TI - Molecular biology. Signal processing in single cells. PMID- 15790835 TI - Atmospheric science. Something in the air. PMID- 15790836 TI - Evolution. The synthesis and evolution of a supermodel. PMID- 15790838 TI - The dynamic gut. PMID- 15790839 TI - What's eating you? PMID- 15790840 TI - A mouthful of microbes. PMID- 15790841 TI - No organ left behind: tales of gut development and evolution. AB - The function of an organ is dependent on its cellular constituents as well as on their assembly into a cohesive unit. The developing gut faces unique challenges as one of the longest and largest organs in the body and also because it is constantly interfacing with external factors through the diet. Its location deep within the body has until recently hampered investigation into its formation. The patterning of the gut along its longitudinal, dorsoventral, left-right, and radial axes is one of the fascinating issues that pertain to the development, function, and homeostasis of this understudied organ. PMID- 15790842 TI - Self-renewal and cancer of the gut: two sides of a coin. AB - The intestinal epithelium follows the paradigms of stem cell biology established for other self-renewing tissues. With a unique topology, it constitutes a two dimensional structure folded into valleys and hills: the proliferative crypts and the differentiated villi. Its unprecedented self-renewal rate appears reflected in a high susceptibility to malignant transformation. The molecular mechanisms that control homeostatic self-renewal and those that underlie colorectal cancer are remarkably symmetrical. Here, we discuss the biology of the intestinal epithelium, emphasizing the roles played by Wnt, bone morphogenic protein, and Notch signaling cascades in epithelial self-renewal and cancer. PMID- 15790843 TI - The gut and energy balance: visceral allies in the obesity wars. AB - In addition to digesting and assimilating nutrients, the intestine and associated visceral organs play a key sensing and signaling role in the physiology of energy homeostasis. The gut, the pancreatic islets of Langerhans, elements in the portal vasculature, and even visceral adipose tissue communicate with the controllers of energy balance in the brain by means of neural and endocrine pathways. Signals reflecting energy stores, recent nutritional state, and other parameters are integrated in the central nervous system, particularly in the hypothalamus, to coordinate energy intake and expenditure. Our understanding of regulatory neural circuits and the signaling molecules that influence them has progressed rapidly, particularly after the discovery of the adipocyte hormone leptin. These discoveries have led to exploration of novel routes for obesity control, some of which involve gut-derived pathways. PMID- 15790844 TI - Host-bacterial mutualism in the human intestine. AB - The distal human intestine represents an anaerobic bioreactor programmed with an enormous population of bacteria, dominated by relatively few divisions that are highly diverse at the strain/subspecies level. This microbiota and its collective genomes (microbiome) provide us with genetic and metabolic attributes we have not been required to evolve on our own, including the ability to harvest otherwise inaccessible nutrients. New studies are revealing how the gut microbiota has coevolved with us and how it manipulates and complements our biology in ways that are mutually beneficial. We are also starting to understand how certain keystone members of the microbiota operate to maintain the stability and functional adaptability of this microbial organ. PMID- 15790845 TI - Immunity, inflammation, and allergy in the gut. AB - The gut immune system has the challenge of responding to pathogens while remaining relatively unresponsive to food antigens and the commensal microflora. In the developed world, this ability appears to be breaking down, with chronic inflammatory diseases of the gut commonplace in the apparent absence of overt infections. In both mouse and man, mutations in genes that control innate immune recognition, adaptive immunity, and epithelial permeability are all associated with gut inflammation. This suggests that perturbing homeostasis between gut antigens and host immunity represents a critical determinant in the development of gut inflammation and allergy. PMID- 15790846 TI - Underwater bipedal locomotion by octopuses in disguise. AB - Here we report bipedal movement with a hydrostatic skeleton. Two species of octopus walk on two alternating arms using a rolling gait and appear to use the remaining six arms for camouflage. Octopus marginatus resembles a coconut, and Octopus (Abdopus) aculeatus, a clump of floating algae. Using underwater video, we analyzed the kinematics of their strides. Each arm was on the sand for more than half of the stride, qualifying this behavior as a form of walking. PMID- 15790847 TI - Widespread parallel evolution in sticklebacks by repeated fixation of Ectodysplasin alleles. AB - Major phenotypic changes evolve in parallel in nature by molecular mechanisms that are largely unknown. Here, we use positional cloning methods to identify the major chromosome locus controlling armor plate patterning in wild threespine sticklebacks. Mapping, sequencing, and transgenic studies show that the Ectodysplasin (EDA) signaling pathway plays a key role in evolutionary change in natural populations and that parallel evolution of stickleback low-plated phenotypes at most freshwater locations around the world has occurred by repeated selection of Eda alleles derived from an ancestral low-plated haplotype that first appeared more than two million years ago. Members of this clade of low plated alleles are present at low frequencies in marine fish, which suggests that standing genetic variation can provide a molecular basis for rapid, parallel evolution of dramatic phenotypic change in nature. PMID- 15790848 TI - Temporal relationships of carbon cycling and ocean circulation at glacial boundaries. AB - Evidence from high-sedimentation-rate South Atlantic deep-sea cores indicates that global and Southern Ocean carbon budget shifts preceded thermohaline circulation changes during the last ice age initiation and termination and that these were preceded by ice-sheet growth and retreat, respectively. No consistent lead-lag relationships are observed during abrupt millennial warming events during the last ice age, allowing for the possibility that ocean circulation triggered some millenial climate changes. At the major glacial-interglacial transitions, the global carbon budget and thermohaline ocean circulation responded sequentially to the climate changes that forced the growth and decline of continental ice sheets. PMID- 15790850 TI - Chemical detection with a single-walled carbon nanotube capacitor. AB - We show that the capacitance of single-walled carbon nanotubes (SWNTs) is highly sensitive to a broad class of chemical vapors and that this transduction mechanism can form the basis for a fast, low-power sorption-based chemical sensor. In the presence of a dilute chemical vapor, molecular adsorbates are polarized by the fringing electric fields radiating from the surface of a SWNT electrode, which causes an increase in its capacitance. We use this effect to construct a high-performance chemical sensor by thinly coating the SWNTs with chemoselective materials that provide a large, class-specific gain to the capacitance response. Such SWNT chemicapacitors are fast, highly sensitive, and completely reversible. PMID- 15790851 TI - Light scattering to determine the relative phase of two Bose-Einstein condensates. AB - We demonstrated an experimental technique based on stimulated light scattering to continuously sample the relative phase of two spatially separated Bose-Einstein condensates of atoms. The phase measurement process created a relative phase between two condensates with no initial phase relation, read out the phase, and monitored the phase evolution. This technique was used to realize interferometry between two trapped Bose-Einstein condensates without need for splitting or recombining the atom cloud. PMID- 15790849 TI - A new population of very high energy gamma-ray sources in the Milky Way. AB - Very high energy gamma-rays probe the long-standing mystery of the origin of cosmic rays. Produced in the interactions of accelerated particles in astrophysical objects, they can be used to image cosmic particle accelerators. A first sensitive survey of the inner part of the Milky Way with the High Energy Stereoscopic System (HESS) reveals a population of eight previously unknown firmly detected sources of very high energy gamma-rays. At least two have no known radio or x-ray counterpart and may be representative of a new class of "dark" nucleonic cosmic ray sources. PMID- 15790852 TI - Cool La Nina during the warmth of the Pliocene? AB - The role of El Nino-Southern Oscillation (ENSO) in greenhouse warming and climate change remains controversial. During the warmth of the early-mid Pliocene, we find evidence for enhanced thermocline tilt and cold upwelling in the equatorial Pacific, consistent with the prevalence of a La Nina-like state, rather than the proposed persistent warm El Nino-like conditions. Our Pliocene paleothermometer supports the idea of a dynamic "ocean thermostat" in which heating of the tropical Pacific leads to a cooling of the east equatorial Pacific and a La Nina like state, analogous to observations of a transient increasing east-west sea surface temperature gradient in the 20th-century tropical Pacific. PMID- 15790853 TI - Soft-tissue vessels and cellular preservation in Tyrannosaurus rex. AB - Soft tissues are preserved within hindlimb elements of Tyrannosaurus rex (Museum of the Rockies specimen 1125). Removal of the mineral phase reveals transparent, flexible, hollow blood vessels containing small round microstructures that can be expressed from the vessels into solution. Some regions of the demineralized bone matrix are highly fibrous, and the matrix possesses elasticity and resilience. Three populations of microstructures have cell-like morphology. Thus, some dinosaurian soft tissues may retain some of their original flexibility, elasticity, and resilience. PMID- 15790854 TI - Glycan foraging in vivo by an intestine-adapted bacterial symbiont. AB - Germ-free mice were maintained on polysaccharide-rich or simple-sugar diets and colonized for 10 days with an organism also found in human guts, Bacteroides thetaiotaomicron, followed by whole-genome transcriptional profiling of bacteria and mass spectrometry of cecal glycans. We found that these bacteria assembled on food particles and mucus, selectively induced outer-membrane polysaccharide binding proteins and glycoside hydrolases, prioritized the consumption of liberated hexose sugars, and revealed a capacity to turn to host mucus glycans when polysaccharides were absent from the diet. This flexible foraging behavior should contribute to ecosystem stability and functional diversity. PMID- 15790855 TI - Introduced predators transform subarctic islands from grassland to tundra. AB - Top predators often have powerful direct effects on prey populations, but whether these direct effects propagate to the base of terrestrial food webs is debated. There are few examples of trophic cascades strong enough to alter the abundance and composition of entire plant communities. We show that the introduction of arctic foxes (Alopex lagopus) to the Aleutian archipelago induced strong shifts in plant productivity and community structure via a previously unknown pathway. By preying on seabirds, foxes reduced nutrient transport from ocean to land, affecting soil fertility and transforming grasslands to dwarf shrub/forb dominated ecosystems. PMID- 15790856 TI - Gene regulation at the single-cell level. AB - The quantitative relation between transcription factor concentrations and the rate of protein production from downstream genes is central to the function of genetic networks. Here we show that this relation, which we call the gene regulation function (GRF), fluctuates dynamically in individual living cells, thereby limiting the accuracy with which transcriptional genetic circuits can transfer signals. Using fluorescent reporter genes and fusion proteins, we characterized the bacteriophage lambda promoter P(R) in Escherichia coli. A novel technique based on binomial errors in protein partitioning enabled calibration of in vivo biochemical parameters in molecular units. We found that protein production rates fluctuate over a time scale of about one cell cycle, while intrinsic noise decays rapidly. Thus, biochemical parameters, noise, and slowly varying cellular states together determine the effective single-cell GRF. These results can form a basis for quantitative modeling of natural gene circuits and for design of synthetic ones. PMID- 15790857 TI - Noise propagation in gene networks. AB - Accurately predicting noise propagation in gene networks is crucial for understanding signal fidelity in natural networks and designing noise-tolerant gene circuits. To quantify how noise propagates through gene networks, we measured expression correlations between genes in single cells. We found that noise in a gene was determined by its intrinsic fluctuations, transmitted noise from upstream genes, and global noise affecting all genes. A model was developed that explains the complex behavior exhibited by the correlations and reveals the dominant noise sources. The model successfully predicts the correlations as the network is systematically perturbed. This approach provides a step toward understanding and manipulating noise propagation in more complex gene networks. PMID- 15790858 TI - RNA-dependent cysteine biosynthesis in archaea. AB - Several methanogenic archaea lack cysteinyl-transfer RNA (tRNA) synthetase (CysRS), the essential enzyme that provides Cys-tRNA(Cys) for translation in most organisms. Partial purification of the corresponding activity from Methanocaldococcus jannaschii indicated that tRNA(Cys) becomes acylated with O phosphoserine (Sep) but not with cysteine. Further analyses identified a class II type O-phosphoseryl-tRNA synthetase (SepRS) and Sep-tRNA:Cys-tRNA synthase (SepCysS). SepRS specifically forms Sep-tRNA(Cys), which is then converted to Cys tRNA(Cys) by SepCysS. Comparative genomic analyses suggest that this pathway, encoded in all organisms lacking CysRS, can also act as the sole route for cysteine biosynthesis. This was proven for Methanococcus maripaludis, where deletion of the SepRS-encoding gene resulted in cysteine auxotrophy. As the conversions of Sep-tRNA to Cys-tRNA or to selenocysteinyl-tRNA are chemically analogous, the catalytic activity of SepCysS provides a means by which both cysteine and selenocysteine may have originally been added to the genetic code. PMID- 15790859 TI - Structural insights into the activity of enhancer-binding proteins. AB - Activators of bacterial sigma54-RNA polymerase holoenzyme are mechanochemical proteins that use adenosine triphosphate (ATP) hydrolysis to activate transcription. We have determined by cryogenic electron microscopy (cryo-EM) a 20 angstrom resolution structure of an activator, phage shock protein F [PspF(1 275)], which is bound to an ATP transition state analog in complex with its basal factor, sigma54. By fitting the crystal structure of PspF(1-275) at 1.75 angstroms into the EM map, we identified two loops involved in binding sigma54. Comparing enhancer-binding structures in different nucleotide states and mutational analysis led us to propose nucleotide-dependent conformational changes that free the loops for association with sigma54. PMID- 15790860 TI - Early lung cancer detection using spiral computed tomography and positron emission tomography. AB - RATIONALE: Lung cancer screening using computed tomography (CT) is effective in detecting lung cancer in early stages. Concerns regarding false-positive rates and unnecessary invasive procedures have been raised. OBJECTIVE: To study the efficiency of a lung cancer protocol using spiral CT and F-18-fluorodeoxyglucose positron emission tomography (FDG-PET). METHODS: High-risk individuals underwent screening with annual spiral CTs. Follow-up CTs were done for noncalcified nodules of 5 mm or greater, and FDG-PET was done for nodules 10 mm or larger or smaller (> 7 mm), growing nodules. RESULTS: A total of 911 individuals completed a baseline CT study and 424 had at least one annual follow-up study. Of the former, 14% had noncalcified nodules of 5 mm or larger, and 3.6% had nodules of 10 mm or larger. Eleven non-small cell lung cancers (NSCLC) and one small cell lung cancer (SCLC) were diagnosed in the baseline study (prevalence rate, 1.32%), and two NSCLCs in the annual study (incidence rate, 0.47%). All NSCLCs (92% of prevalence cancers) were diagnosed in stage I (12 stage IA, 1 stage IB). FDG-PET was helpful for the correct diagnosis in 19 of 25 indeterminate nodules. The sensitivity, specificity, positive predictive value, and negative predictive value of FDG-PET for the diagnosis of malignancy were 69, 91, 90, and 71%, respectively. However, the sensitivity and negative predictive value of the screening algorithm, which included a 3-month follow-up CT for nodules with a negative FDG-PET, was 100%. CONCLUSION: A protocol for early lung cancer detection using spiral CT and FDG-PET is useful and may minimize unnecessary invasive procedures for benign lesions. PMID- 15790861 TI - The role of hyaluronan synthase 3 in ventilator-induced lung injury. AB - We recently found that low-molecular-weight hyaluronan was induced by cyclic stretch in lung fibroblasts and accumulated in lungs from animals with ventilator induced lung injury. The low-molecular-weight hyaluronan produced by stretch increased interleukin-8 production in epithelial cells, and was accompanied by an upregulation of hyaluronan synthase-3 mRNA. We hypothesized that low-molecular weight hyaluronan induced by high VT was dependent on hyaluronan synthase 3, and was associated with ventilator-induced lung injury. Effects of high VT ventilation in C57BL/6 wild-type and hyaluronan synthase-3 knockout mice were compared. Significantly increased neutrophil infiltration, macrophage inflammatory protein-2 production, and lung microvascular leak were found in wild type animals ventilated with high VT. These reactions were significantly reduced in hyaluronan synthase-3 knockout mice, except the capillary leak. Wild-type mice ventilated with high VT were found to have increased low-molecular-weight hyaluronan in lung tissues and concomitant increased expression of hyaluronan synthase-3 mRNA, neither of which was found in hyaluronan synthase-3 knockout mice. We conclude that high VT induced low-molecular-weight hyaluronan production is dependent on de novo synthesis through hyaluronan synthase 3, and plays a role in the inflammatory response of ventilator-induced lung injury. PMID- 15790862 TI - The NIH Public Access Policy and ATS journals. PMID- 15790863 TI - Airway remodeling: therapeutic target or not? PMID- 15790864 TI - Lung allograft ischemic time: crossing the threshold. PMID- 15790865 TI - Interstitial lung disease, lung cancer, lung transplantation, pulmonary vascular disorders, and sleep-disordered breathing in AJRCCM in 2004. PMID- 15790866 TI - Allergic rhinitis, asthma, airway biology, and chronic obstructive pulmonary disease in AJRCCM in 2004. PMID- 15790867 TI - Lesions caused by ricin applied to rabbit eyes. AB - PURPOSE: Ricin, a highly potent toxin from castor beans, is a potential biological weapon that could be dispersed in the air as dust or aerosol. In these forms, ricin, besides being inhaled, could reach unprotected eyes. The present research was performed to ascertain the lesions that the toxin causes when applied to rabbit eyes. METHODS: Ricin was applied to rabbit eyes in solution, in quantities ranging from 1 to 100 mug. Animals were observed until death, when eyes and internal organs were removed and fixed. Sections were stained and examined microscopically. RESULTS: Ricin caused inflammation of the eyes and adnexa, visible both macroscopically and histologically. The damage was greatly reduced by rinsing the eyes with 10% lactose, provided the rinsing was done almost immediately after application of the toxin. Rinsing with phosphate buffered saline (PBS) had no effect. With the highest dosage, congestion of internal organs was also apparent. CONCLUSIONS: Application of ricin to eyes causes local damage, mainly of the inflammatory type. The ineffectiveness of rapid rinsing with PBS and the partial efficacy of rapid rinsing with lactose indicate that the toxin quickly binds to and is taken up by cells. The lesions of internal organs show that ricin applied to the eyes can be absorbed, pass into the circulation, and, at least at some dosages, damage internal organs. PMID- 15790868 TI - Light-induced oxidative stress in choroidal endothelial cells in mice. AB - PURPOSE: Although light-induced oxidative stress in the retina has been extensively reported, little information regarding light-induced oxidative stress in choroidal endothelial cells (CECs) is available. In the current study, light induced DNA oxidation and the activation of nuclear factor-kappaB (NF-kappaB), a major oxidative responsive transcription factor, were investigated in mouse CECs. METHODS: Mice were exposed to green light. Light-induced DNA oxidation in CECs was detected by in situ 8-hydroxy-2-deoxyguanosine (8-oxo-dG) immunolabeling. CECs were isolated from the retinal pigment epithelium (RPE)/choroid by using immunomagnetic beads. The isolated CECs were immunochemically characterized by the expression of endothelial markers, CD31, and P1H12. The quality of total RNA from CECs was assessed by a bioanalyzer and RT-PCR. NF-kappaB activation in situ and in isolated CECs was investigated. RESULTS: After a 3-hour exposure to light, the immunoreactivity to anti-8-oxo-dG antibody or anti-NF-kappaB p65 antibody in CECs in situ was significantly increased when compared with unexposed mice. Isolated CECs expressed CD31 and P1H12. The 28S/18S rRNA ratio of RNA isolated from CECs was 1.5:1. CD31 and von Willebrand Factor (vWF) transcripts were predominantly expressed in the RNA from isolated CECs. IkappaBalpha was more heavily phosphorylated in light-exposed than untreated CECs. IkappaBalpha expression levels were increased fivefold in isolated CECs after exposure to light compared to unexposed control subjects. CONCLUSIONS: Exposure to light induces oxidative stress in CECs in vivo. A method for CEC isolation from the mouse RPE/choroid with preservation of RNA quality has been developed. The results of this study may facilitate the ability to identify CEC-specific genes and gene products that respond to photo-oxidative stress. PMID- 15790870 TI - Hereditary lattice corneal dystrophy is associated with corneal amyloid deposits enclosing C-terminal fragments of keratoepithelin. AB - PURPOSE: To investigate the molecular basis of hereditary lattice corneal dystrophy (LCD) type IIIA associated with corneal amyloid deposits afflicting several members of a four-generation family. METHODS: Histologic, immunohistochemical and biochemical studies were performed on corneal tissue samples obtained after perforating keratoplasty. DNA was extracted from peripheral blood leukocytes. All exons of the keratoepithelin-encoding TGFBI gene were amplified and sequenced. The presence of a mutation was confirmed by digestion of the isolated PCR product with the restriction enzyme AlwNI. RESULTS: The cornea of the index patient (II-1) contained large patchy deposits of amyloid, which were immunoreactive for the C terminus of keratoepithelin. Western blot analysis of the polypeptide chains extracted from the amyloid deposits of paraffin-embedded tissue revealed that these represented mainly fragments of the full-length protein. The smallest fragments were 6.5 and 6.9 kDa. DNA analyses of the TGFBI gene revealed a heterozygous T-->C transition at the second position of codon 540 in exon 12, indicating that replacement of phenylalanine by serine (Phe540Ser) leads to dominant disease. The mutation creates a new restriction site for the enzyme AlwNI. Five of the examined family members carried this mutation. Three of them (aged >/=41 years) had the disease, two family members (aged <20 years) do not yet show any clinical symptoms. An additional inconsequential single-nucleotide polymorphism (T1667C) was found at the third position of the same codon (Phe540Phe) in three unaffected family members. CONCLUSIONS: This is the first report of a single-nucleotide mutation at codon 540 of TGFBI leading to LCD, and the first to demonstrate that the amyloid deposits in LCD contain proteolytic fragments of keratoepithelin. PMID- 15790869 TI - A novel GCAP1 missense mutation (L151F) in a large family with autosomal dominant cone-rod dystrophy (adCORD). AB - PURPOSE: To elucidate the phenotypic and biochemical characteristics of a novel mutation associated with autosomal dominant cone-rod dystrophy (adCORD). METHODS: Twenty-three family members of a CORD pedigree underwent clinical examinations, including visual acuity tests, standardized full-field ERG, and fundus photography. Genomic DNA was screened for mutations in GCAP1 exons using DNA sequencing and single-strand conformational polymorphism (SSCP) analysis. Function and stability of recombinant GCAP1-L151F were tested as a function of [Ca(2+)], and its structure was probed by molecular dynamics. RESULTS: Affected family members experienced dyschromatopsia, hemeralopia, and reduced visual acuity by the second to third decade of life. Electrophysiology revealed a nonrecordable photopic response with later attenuation of the scotopic response. Affected family members harbored a C-->T transition in exon 4 of the GCAP1 gene, resulting in an L151F missense mutation affecting the EF hand motif 4 (EF4). This change was absent in 11 unaffected family members and in 100 unrelated normal subjects. GCAP1-L151F stimulation of photoreceptor guanylate cyclase was not completely inhibited at high physiological [Ca(2+)], consistent with a lowered affinity for Ca(2+)-binding to EF4. CONCLUSIONS: A novel L151F mutation in the EF4 hand domain of GCAP1 is associated with adCORD. The clinical phenotype is characterized by early cone dysfunction and a progressive loss of rod function. The biochemical phenotype is best described as persistent stimulation of photoreceptor guanylate cyclase, representing a gain of function of mutant GCAP1. Although a conservative substitution, molecular dynamics suggests a significant change in Ca(2+)-binding to EF4 and EF2 and changes in the shape of L151F-GCAP1. PMID- 15790871 TI - Vitamins C and e reduce retinal oxidative stress and nitric oxide metabolites and prevent ultrastructural alterations in porcine hypercholesterolemia. AB - PURPOSE: Oxidative stress is thought to be involved in the pathogenesis of age related diseases, such as atherosclerosis and retinal degeneration. The current study was conducted to examine vitreoretinal oxidative status in a model of porcine hypercholesterolemia to identify morphologic alterations and analyze the effect of dietary supplementation with vitamins C and E. METHODS: Adult miniature pigs were fed standard chow, cholesterol-rich chow, or a cholesterol-rich diet supplemented with vitamins C and E. Total cholesterol, triglycerides, high density lipoproteins, lipid peroxidation, and tocopherol were measured in plasma. Lipid peroxidation and nitric oxide (NO) metabolites were measured in vitreous and retinal homogenates. Superoxide anion release in the retinal pigment epithelium (RPE) was analyzed by chemiluminescence. Retinal morphology was studied by transmission electron microscopy. RESULTS: The high-cholesterol group, with increased retinal oxidative stress (P < 0.01) and NO metabolites in the retina (P < 0.05), had increased superoxide anion release (P < 0.05) and showed development of pyknosis, irregular nuclear membranes, and cytoplasmic accumulation of lipids and autophagocytic vacuoles in the RPE cells. Vitamins C and E prevented biochemical changes and most ultrastructural alterations in the RPE. CONCLUSIONS: The results suggest an evolving role for hypercholesterolemia through increased retinal oxidative stress and NO synthesis that could be responsible for retinal ultrastructural alterations. The beneficial effects of vitamins C and E in the retinal tissue further support this hypothesis. PMID- 15790872 TI - Detailed characterization of the lipid composition of detergent-resistant membranes from photoreceptor rod outer segment membranes. AB - PURPOSE: In recent years, detergent-resistant membranes (DRMs) have been isolated in in vitro models of lipid rafts, from photoreceptor outer segments (ROS), and the localization of a specific complement of photoreceptor proteins has been demonstrated. However, surprisingly little is known about the lipid composition of these important membrane domains. The present study provides the first characterization of phospholipids and fatty acids from ROS-derived DRMs. METHODS: Bovine ROS membranes were incubated with 1% Triton X-100 at 4 degrees C and subjected to density gradient centrifugation to isolate DRMs from the parent membranes. Lipids of ROS and DRMs were separated by two-dimensional, thin-layer chromatography and converted to methyl esters, and fatty acids were analyzed by gas chromatography. Proteins of ROS and DRMs were analyzed by SDS-PAGE and Western blot analysis. RESULTS: The DRMs represented 8% and 3%, respectively, of total ROS lipid and protein. In general, DRMs were enriched in saturated fatty acids when compared with ROS membranes. Relative to ROS, DRMs were enriched in free fatty acids (FFAs) and a specific phosphatidylcholine (PC) fraction that was almost devoid of polyunsaturated fatty acids (PUFAs). DRMs contained less phosphatidylethanolamine (PE) and phosphatidylserine (PS). Ceramide (CM) from ROS contained PUFAs but no saturated fatty acids; the converse was true of CM from DRMs. Docosahexaenoic acid was diminished in DRM PS and was not detected in the FFAs, but was equally abundant in ROS and DRM PE. ROS-derived DRMs were dramatically enriched in caveolin-1, contained significant amounts of transducin alpha and c-Src, and were relatively devoid of arrestin. CONCLUSIONS: The relatively saturated lipid environment observed in DRMs is likely to promote the localization of signaling proteins modified with saturated fatty acyl chains. Based on the lipid composition of DRMs, the authors conclude that they would not efficiently support phototransduction. PMID- 15790873 TI - Detecting progression of nuclear sclerosis by using human grading versus semiautomated computer grading. AB - PURPOSE: To assess indices of nuclear sclerosis derived from digitized images made from color (slide) photographs. METHODS: Film-based slit lamp images taken at baseline and at 5- and 10-year follow-up examinations of the Beaver Dam Eye Study cohort were digitized, and optical traces were taken along an axis through the center of the cornea and lens. Four indices of the severity of sclerosis were calculated based on the optical densities. The associations of the original Beaver Dam grades and these indices to age, vision, and change in severity of sclerosis over two subsequent visits were compared. RESULTS: At baseline photographs, the Spearman correlation between age and severity was 0.65 for the original film-based grading (n = 4518 right eyes) and varied between 0.46 and 0.71 for the measures from digitized images. Correlations of the indices to visual acuity were 0.38 for the film-based grading and ranged from 0.32 to 0.38 for the other indices. The authors assume that nuclear sclerosis does not regress and the percentage of regression is a reflection of error in grading. The percentage of regression and progression of sclerosis over 5- and 10-year intervals was determined for each index. After 5 years, 48.2% progressed and 4.9% regressed, using the Beaver Dam grades; progression occurred in 4.9% to 9.9%, and regression occurred in 4.5% to 7.0% for the other indices. After 10 years, 61.9% progressed and 3.2% regressed using the Beaver Dam grades; progression occurred in 8.0% to 19.7%, and regression occurred in 2.6% to 9.7% for the other indices. CONCLUSIONS: Semiautomated grading of the digitized images can be used to process thousands of images with little oversight by a trained grader. Indices of sclerosis that closely parallel human grading in their relationships to age and visual acuity can be easily computed. However, the indices appear to identify significantly less progression of nuclear sclerosis than does human grading. Further development to define a useful metric for identifying severity and progression of nuclear sclerosis is needed. PMID- 15790874 TI - Ocular growth in premature infants conceived by in vitro fertilization versus natural conception. AB - PURPOSE: To evaluate the possible effect of in vitro fertilization (IVF) on early development of the eye in premature infants. METHODS: Biometric and keratometric values, intraocular pressure, and retinal vascular status were assessed in a cohort of 133 premature infants. These values were compared between premature infants conceived by IVF or naturally, and the relationship between these parameters and postconceptional age and weight at examination were evaluated. RESULTS: The sample consisted of 133 premature infants, 62 (46.6%) born by IVF and 71 (53.4%) by natural conception. Postconceptional age at examination was 28 to 46 weeks. In both groups, axial length, anterior chamber depth, and corneal radius correlated with the postconceptional age and weight at examination and followed a linear growth pattern. Lens thickness changed very slightly. The rate of retinal vascularization correlated with the postconceptional age as well. No correlation was found between intraocular pressure and corrected age or weight at examination. There was no difference between the study and control groups in any of the biometric or keratometric parameters or in intraocular pressure, according to two-way analysis of variance. CONCLUSIONS: IVF apparently does not affect early ocular growth, intraocular pressure, changes in corneal curvature, or retinal vascularization in premature infants. These findings may aid ophthalmologists in assessing ocular dimensions in this patient population. PMID- 15790875 TI - Pseudoexfoliation in southern India: the Andhra Pradesh Eye Disease Study. AB - PURPOSE: To report the prevalence of pseudoexfoliation (PXF) and its associations with ocular diseases in a south Indian population. METHODS: This was a population based, cross-sectional epidemiologic study in the south Indian state of Andhra Pradesh (AP). A total of 10,293 subjects of all ages from one urban and three rural areas representative of the population of AP were interviewed and underwent a comprehensive ophthalmic evaluation. PXF was diagnosed on slit lamp biomicroscopy by the presence of white dandruff-like material in the pupillary margin, on the trabecular meshwork, and/or on the anterior lens capsule of one or both eyes. RESULTS: The age-gender-area-adjusted overall prevalence of PXF was 0.69% (95% CI: 0.53-0.86). The prevalence of PXF increased with increasing age: 3.01% (95% CI: 2.45-3.80), in those 40 years of age or older, and 6.28% (95% CI: 4.80-7.76), in those 60 years of age or older. The prevalence of PXF was significantly higher among people whose occupation involved outdoor activities (adjusted odds ratio [OR], 2.14; 95% CI: 1.10-4.16). After adjustment for age, the prevalence of PXF was significantly higher in those with nuclear cataract (adjusted OR, 2.00; 95% CI: 1.13-3.54). PXF was significantly associated with blindness (adjusted OR, 2.19; 95% CI: 1.16-4.13). Fifteen (20.5%; 95% CI: 11.20 29.80) of those with PXF were blind, with age-adjusted relative risk (RR) = 4.25 (95% CI: 4.01-4.51). Unilateral blindness (41.2%; 95% CI: 29.81-52.39) and visual impairment (45.21%; 95% CI: 34.29-57.13) were also more common with PXF. Four subjects (5.5%; 95% CI: 0.27-10.2) of those with PXF had glaucoma. The prevalence of PXF in those with glaucoma was 4.2%; (95% CI: 0.17-8.23). In general linear models, the estimated mean +/- SE of IOP with glaucoma and PXF was 24.14 +/- 1.41 mm Hg and was 18.94 +/- 0.26 mm Hg with glaucoma in the absence of PXF; the difference was statistically significant (P < 0.0001). CONCLUSIONS: The association of PXF with blindness and aging has public health implications for India. This is especially so considering the burden of cataract with aging and the association of PXF with cataract as well as complications of cataract surgery. The diagnosis of PXF may also be important in the management of glaucoma in this population. PMID- 15790876 TI - Common visual defects and peer victimization in children. AB - PURPOSE: To investigate whether wearing glasses, having manifest strabismus, or having a history of wearing an eye patch predisposes preadolescent children to being victimized more frequently at school and whether the impact may be different on boys than on girls. METHODS: Data were examined on 6536 children from the Avon Longitudinal Study of Parents and Children (ALSPAC) based in the United Kingdom. At 7.5 years, the children undertook a detailed eye examination by orthoptists, including a cover test and visual acuity assessment. At 8.5 years, trained psychologists assessed the children's bullying involvement as either victim or perpetrator for overt and relational bullying, in a standard interview. RESULTS: Children currently wearing glasses or with a history of wearing eye patches were 35% to 37% more likely to be victims of physical or verbal bullying, even after adjustment for social class and maternal education. No interactions were found between sex and visual problems in the prediction of bullying. CONCLUSIONS: For those children who require glasses, opticians should be aware of the risks of bullying, and strategies should be developed and discussed that help reduce their vulnerability. PMID- 15790877 TI - Retinal vessel diameters and incident open-angle glaucoma and optic disc changes: the Rotterdam study. AB - PURPOSE: It remains unclear whether reduced retinal blood flow and smaller arterioles, reported to exist in patients with open-angle glaucoma (OAG), are a cause or a consequence of ganglion cell loss. We examined whether baseline retinal vessel diameters were related to incident (i)OAG or incident optic disc changes in a population-based sample. METHODS: In the prospective population based Rotterdam Study, baseline diameters of retinal arterioles and venules (1990 1993) were measured in digitized images of 3469 persons (aged 55 years and older) at risk for OAG. The follow-up examinations took place from 1997 to 1999. iOAG was based on the presence of incident glaucomatous visual field loss and/or incident glaucomatous optic neuropathy. Changes in neuroretinal rim, cup area, or vertical cup-to-disc ratio were calculated with a semiautomated image analyzer in 2782 persons. RESULTS: After a mean follow-up time of 6.5 years, 74 participants had iOAG. At baseline, the mean arteriolar diameter was 147.5 +/- 14.2 microm (SD) and the venular, 222.9 +/- 20.0 microm. Neither arteriolar diameters (odds ratio [OR] per SD decrease: 0.82; 95% confidence interval [CI]: 0.66-1.03) nor venular ones (OR per SD increase: 1.20; 95% CI: 0.95-1.53) were significantly related to iOAG. Baseline retinal vessel diameters did not predict changes in the optic disc. Additional adjustment for cardiovascular risk factors did not alter these results. CONCLUSIONS: The data show that baseline retinal vessel diameters did not influence the risk of iOAG or incident optic disc changes. These data provide no evidence for a retinal vascular role in the pathogenesis of OAG. PMID- 15790878 TI - Tear film dynamics in floppy eyelid syndrome. AB - PURPOSE: Floppy eyelid syndrome (FES) presents nonspecific ocular surface irritation. The hypothesis for the current study was that one contributing factor is the abnormality in tear film dynamics. METHODS: Sixteen patients with FES were consecutively examined. Tear film dynamics were evaluated by kinetic tear interference images, infrared thermometry, water evaporation rate, tear break-up time, and fluorescein clearance test. Data showing evaporation rate and thermometry were compared with those of 10 normal subjects. RESULTS: There was a high correlation between the eye with the worse symptoms and the eyes with the more severe floppy lids (P < 0.01) and with ocular surface evaporation rate (P = 0.02). Except for one patient, all others showed abnormal tear film, with an average tear break-up time of 2.9 +/- 3.7 seconds. Kinetic analysis of tear interference images revealed that lipid spread in a vertical or mixed pattern in 18 eyes (75%) with a delayed spread time (P = 0.0007), indicating that most of the patients had lipid tear deficiency. The ocular skin temperature and water evaporation rate were higher in the FES group (P = 0.0003 and 0.026, respectively). Nearly all patients with FES showed eyelid hyperpigmentation. The ocular surface evaporation rate in the FES group was also higher than that of the normal subjects (P < 0.0001). Multiple regression analysis showed that a vertical pattern of lipid spread had a significant influence on ocular surface evaporation rate (P = 0.003). CONCLUSIONS: Tear film abnormality is prevalent in patients with FES and is characterized by lipid tear deficiency, leading to rapid tear evaporation. The FES lid skin is also characterized by high temperature, high water evaporation rate, and hyperpigmentation. Studies directed to investigating the linkage of lid changes and meibomian gland dysfunction may shed new lights on the pathogenesis of FES. PMID- 15790879 TI - Inhibition of gap junction-mediated intercellular communication by TNF-alpha in cultured human corneal fibroblasts. AB - PURPOSE: Keratocytes are connected to each other by gap junctions, which mediate intercellular communication and contribute to maintenance of corneal homeostasis. The possible effect of tumor necrosis factor (TNF)-alpha, a proinflammatory cytokine, on gap junctional intercellular communication (GJIC) in cultured human corneal fibroblasts was examined. METHODS: GJIC activity was measured by observing the intercellular diffusion of the fluorescent dye Lucifer yellow. The expression of the gap junction protein connexin43 (Cx43) was evaluated by immunofluorescence and immunoblot analyses with a specific monoclonal antibody. The abundance of Cx43 mRNA was determined by quantitative reverse transcription and polymerase chain reaction analysis. RESULTS: TNF-alpha induced a time- and concentration-dependent decrease in GJIC activity in human corneal fibroblasts. Immunofluorescence analysis revealed that TNF-alpha reduced the level of specific staining for Cx43 at sites of contact between adjacent cells. Immunoblot analysis detected four specific Cx43 bands, one corresponding to the nonphosphorylated form of the protein and three corresponding to phosphorylated forms. Exposure of cells to TNF-alpha reduced the relative abundance of the three phosphorylated forms of Cx43. The amount of Cx43 mRNA was not affected by TNF-alpha. CONCLUSIONS: TNF-alpha inhibited GJIC in cultured human corneal fibroblasts, an effect that was possibly mediated by dephosphorylation and consequent degradation of Cx43. The downregulation of GJIC among keratocytes in response to TNF-alpha may contribute to the breakdown of corneal homeostasis during corneal inflammation. PMID- 15790880 TI - CCR5 chemokine receptor mediates recruitment of MHC class II-positive Langerhans cells in the mouse corneal epithelium. AB - PURPOSE: To characterize the chemokines and chemokine receptors that mediate the effect of proinflammatory cytokines, interleukin (IL)-1 and tumor necrosis factor (TNF)-alpha, on the recruitment of MHC class II(+) Langerhans cells (LCs) in the corneal epithelium. METHODS: A standard model for corneal LC recruitment, application of cautery to the central corneal surface was used, and the differential gene expression levels of a panel of chemokines and chemokine receptors were determined by RNase protection assay. Chemokine receptor-knockout mice were used to evaluate the recruitment of MHC class II(+) LCs to the corneal epithelium. To determine the sensitivity of selected chemokines to IL-1 and TNF alpha stimulation, the chemokine gene expression pattern was analyzed after blockade of IL-1 and TNF receptors. RESULTS: CCR1, -2, and -5 were overexpressed in corneas after cauterization. Topical administration of soluble TNF receptor I and IL-1 receptor antagonist, which abrogated corneal LC recruitment, significantly suppressed the gene transcription levels of the ligands of CCR1 and/or -5, regulated on activation normal T-cell expressed and secreted (RANTES), macrophage inflammatory protein (MIP)-1alpha, and MIP-1beta. The recruitment of major histocompatibility complex (MHC) class II(+) LC was significantly suppressed in CCR5(-/-) mice and blockade of RANTES and MIP-1beta, but not in CCR1(-/-), CCR2(-/-)/MIP-1alpha(-/-), or MIP-1alpha(-/-) mice. The evaluation of epithelial CD11c(+) LC cells by confocal microscopy revealed coexpression for CCR5 primarily among B7(-) (CD80(-)/CD86(-)) subsets of these LCs but not among the mature B7(+) subsets of CD11c(+) LCs. CONCLUSIONS: These data suggest that CCR5 plays a critical role in mediating recruitment and mobilization of MHC class II(+) LCs into the corneal epithelium. Targeting CCR5 and its ligands may be a new strategy for modulating immunity. PMID- 15790881 TI - ATP release through connexin hemichannels in corneal endothelial cells. AB - PURPOSE: Intercellular Ca(2+) wave propagation is a distinct form of cell-cell communication. In corneal endothelial cells, intercellular Ca(2+) wave propagation evoked by a point mechanical stimulus (PMS) is partially mediated by adenosine triphosphate (ATP) release and subsequent activation of P2Y receptors. This study was conducted to investigate the possibility that extrajunctional connexons (hemichannels) play a role in ATP release during PMS-induced Ca(2+) wave propagation in bovine corneal endothelial cells (BCECs). METHODS: A Ca(2+) wave was evoked by a PMS applied to a single cell in a monolayer of cultured BCECs. Changes in [Ca(2+)](i) in the mechanically stimulated cell (MS cell) and in the neighboring (NB) cells were visualized by fluorescence imaging using the Ca(2+)-sensitive dye Fluo-4. From these images, the maximum normalized fluorescence (NF), the percentage of responsive cells (%RC), and the total area of cells reached by the Ca(2+) wave (active area [AA], in square micrometers) were calculated. Intercellular dye transfer, generally attributed to gap junctional coupling, was assessed by fluorescence recovery after photobleaching (FRAP) using 6-carboxyfluorescein diacetate. Opening of hemichannels was investigated by measuring cellular uptake of the fluorescent dye Lucifer yellow, which is known to permeate hemichannels. ATP release was measured by luciferin luciferase bioluminescence. RESULTS: Flufenamic acid (FFA; 50 microM) and the connexin mimetic peptide Gap26 (300 microM), known blockers of hemichannels, significantly reduced AA in confluent monolayers as well as in contact-free cells. Neither FFA nor Gap26 affected the FRAP, indicating that reduction in AA of the PMS-induced wave by these agents is not due to a block of gap junction channels. FFA as well as Gap26 inhibited the increase in AA of the wave that was observed when cells were pretreated with the ectonucleotidase inhibitor ARL-67156 (100 microM). These findings suggest that the hemichannel blockers reduce the Ca(2+) wave propagation by inhibiting ATP release. Consistent with this finding, PMS or exposure to Ca(2+)-free solution (a maneuver known to induce the opening of hemichannels) led to ATP release; moreover, the release was inhibited by the hemichannel blockers. The extracellular ATP levels in response to both PMS and extracellular Ca(2+) removal were strongly enhanced by ARL-67156, and this effect was inhibited by FFA as well as by Gap26. Moreover, pretreatment of subconfluent BCEC monolayers with FFA or Gap26 inhibited the uptake of Lucifer yellow induced by removal of extracellular Ca(2+). CONCLUSIONS: Hemichannels contribute to ATP release on mechanical stimulation in BCECs. The released ATP contributes to propagation of the Ca(2+) wave. PMID- 15790882 TI - An involucrin promoter AP1 transcription factor binding site is required for expression of involucrin in the corneal epithelium in vivo. AB - PURPOSE: Cell division of corneal limbal stem cells gives rise to transient amplifying cells that ultimately differentiate to form the multilayered corneal epithelium. The mechanisms that regulate changes in gene expression during this process are not well understood. In the present study, the involucrin gene was used as a model to study this regulation. METHODS: Regulation of human involucrin gene expression and promoter activity was assessed using in vivo transgenic mouse models and cultured primary human corneal epithelial cells. RESULTS: Human involucrin (hINV) is a structural protein that is selectively expressed in differentiating corneal epithelial cells. The results reveal that an activator protein one (AP1) DNA-binding site is essential for appropriate basal and stimulus-dependent hINV promoter activity. Mutation of this site, AP1-5, results in a loss of hINV gene expression in the corneal epithelium in vivo and in cultured corneal epithelial cells. A gel mobility supershift analysis revealed interaction of the AP1 factors, Fra-1 and JunB, with this element. Inhibition of AP1 function with a dominant-negative form of AP1 also inhibited expression. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C activator, increased hINV gene expression, a response that correlates with increased AP1 factor (Fra-1 and JunB) binding to the hINV gene AP1-5 response element. CONCLUSIONS: These findings point to an essential role for AP1 transcription factors, acting through a distal regulatory region AP1-5 element, in the regulation of involucrin gene expression during corneal epithelial cell differentiation. PMID- 15790883 TI - Membrane array characterization of 80 chemokines, cytokines, and growth factors in open- and closed-eye tears: angiogenin and other defense system constituents. AB - PURPOSE: To adapt membrane-bound antibody array (MA) technology to characterize the distribution of a wide range of bioactive trace proteins in reflex (RTF) and open-eye (OTF) and closed-eye (CTF) tear samples. METHODS: Tears were collected by capillary tube and centrifuged. A commercially available standard MA and a custom array were modified to maximize the sensitivity of detection and the signal-to-noise ratio, to assay RTF and individually pooled CTF and OTF samples for 80 chemokines, growth factors, cytokines, and angiogenic modulators. The reliability of data was assessed by Western blot and other methods. RESULTS: Coupling an ultrasensitive chemiluminescence substrate system to an MA and optimizing conditions enhanced the sensitivity several hundredfold, allowing the detection of approximately 40 of the 79 probed proteins on the standard array, most of which were shown to be elevated in CTF. Identified entities include the known constituents epidermal growth factor (EGF), monocyte chemoattractant protein (MCP)-1, IL-8, tissue inhibitor of metalloproteinase (TIMP)-1 and -2, and numerous previously undetected tear components, such as angiogenin (ANG), growth factors, and the CXC and CC chemokines IFN-gamma inducible protein (IP)-10, growth-related oncogene (GRO), epithelial neutrophil-activating protein (ENA)-78, and macrophage inflammatory protein (MIP)-3alpha. Identification of other proteins was hindered by high background on the negative control array. Using a less complex custom array dramatically reduced background and allowed the visualization in CTF of proteins, such as VEGF, that were not detected with the standard array. CONCLUSIONS: MAs are powerful tools for differential screening of tears for large numbers of trace proteins. Analysis allowed the identification of previously undetected proteins that may participate in the host defense system as well as demonstrated the profound change in tear composition associated with prolonged eye closure in a manner reflective of physiological function. PMID- 15790884 TI - Gene expression profile studies of human keratoconus cornea for NEIBank: a novel cornea-expressed gene and the absence of transcripts for aquaporin 5. AB - PURPOSE: To increase the database of genes expressed in human cornea and to gain insights into the molecular basis of keratoconus (KC). METHODS: A cDNA library was constructed from KC corneas harvested at keratoplasty and used for expressed sequence tag (EST) analysis. Data were analyzed using grouping and identification of sequence tags (GRIST). Expression of selected clones was examined by RT-PCR. RESULTS: A total of 7680 clones was sequenced from the 5' end. After bioinformatics analysis, 4090 clusters of clones, each potentially representing individual genes, were identified. Of these, 887 genes were represented by more than one clone. The five most abundant transcripts, represented by >60 clones each, were for keratin-12, TGFBI (BIGH3), decorin, ALDH3, and enolase 1, all known markers for cornea. Many other markers for epithelial, stromal, and endothelial genes were also present. One cluster of six clones came from an apparently novel gene (designated KC6) located on chromosome 18 at p12.3. RT-PCR of RNA from several human tissues detected KC6 transcripts only in cornea. In addition, no clones were observed for the usually prominent corneal epithelial cell marker aquaporin 5 (AQP5), a water channel protein. Semiquantitative RT-PCR confirmed that expression of AQP5 is much lower in KC cornea than in non-KC cornea. CONCLUSIONS: This analysis increases the database of genes expressed in the human cornea and provides insights into KC. KC6 is a novel gene of unknown function that shows cornea-preferred expression, whereas the suppression of transcripts for AQP5 provides the first clear evidence of a molecular defect identified in KC. PMID- 15790885 TI - A modified chondroitin sulfate aldehyde adhesive for sealing corneal incisions. AB - PURPOSE: To compare a modified chondroitin sulfate aldehyde adhesive with standard sutures for sealing corneal incisions. METHODS: A keratome knife was used to create non-self-sealing, uniplanar, 3-mm, clear corneal incisions in enucleated rabbit eyes (n = 18). The wounds were sealed with either a chondroitin sulfate-aldehyde adhesive (n = 8), three 10-0 nylon sutures (n = 5), or one 10-0 nylon suture (n = 5). Wound stability was tested by filling the globes with balanced salt solution through an anterior chamber port and slowly increasing the IOP. The pressure changes were monitored with a digital manometer connected to the anterior chamber, and leak pressure was recorded for each eye. Confocal microscopy was performed on the glued eyes, to document the glue distribution along the wound. RESULTS: The mean leak pressures in the single-suture and three suture subgroups were 26.4 +/- 6.0 and 44.3 +/- 8.2 mm Hg (SD), respectively. The maximum IOP achieved in eyes that received the glue was 104.7 mm Hg with a mean of 101.4 +/- 3.2 mm Hg. None of the eyes in which glue was used showed leakage. At confocal microscopy, the glue was distributed inside the wound edges as a homogeneous thin layer of a less dense signal than that of the stroma. CONCLUSIONS: A novel chondroitin sulfate-aldehyde adhesive was shown to be effective ex vivo for sealing corneal incisions in rabbit eyes and was superior to sutures for this purpose. PMID- 15790886 TI - The inhibitory interaction between human corneal and conjunctival sensory channels. AB - PURPOSE: To explore human corneal and conjunctival sensory channels at suprathreshold level. METHODS: Ten healthy human subjects participated in the study. The Belmonte pneumatic esthesiometer was used to apply mechanical and chemical stimuli to the central cornea and temporal conjunctiva of the left eye. Stimuli were applied in a paired and unpaired way for conjunctival stimulation. A 100-point visual analog scale (VAS) was used to rate the intensity of the stimulus. RESULTS: The magnitudes of the sensation evoked from the conjunctiva were different when using different methods for presenting stimuli to the ocular surface. When stimuli were applied to the conjunctiva alone, the magnitude of the sensation was stronger than when the stimuli were applied in pairs to the cornea and conjunctiva for both mechanical (P = 0.04) and chemical (P = 0.02) stimulation. CONCLUSIONS: The relatively strong discomfort evoked from the cornea appears to suppress partially the relatively weaker conjunctival stimulation. This manifested as the conjunctival sensory transducer function being shallower (less intense sensation) when immediately preceded by corneal stimulation than when the conjunctival sensory transducer functions were measured alone (unpaired). The underlying mechanism could be adaptation or some other inhibitory effect, such as diffuse noxious inhibitory control. At some level therefore, corneal and conjunctival sensory channels are not independent. PMID- 15790887 TI - Accumulation of mitochondrial DNA damage in keratoconus corneas. AB - PURPOSE: To determine whether keratoconus (KC) corneas have more mitochondrial (mt)DNA damage than do normal corneas. METHODS: Thirty-three normal corneas and 34 KC corneas were studied. Immunohistochemistry for mitochondria-encoded cytochrome c oxidase (complex IV) subunit 1 (CO-Iota) and porins was performed. Total DNA was isolated and mtDNA genome amplified by either long-extension polymerase chain reaction (LX-PCR) or short-extension-PCR (SX-PCR). LX-PCR mtDNA was digested with restriction enzymes to confirm full-length mtDNA amplicon. SX PCR mtDNA was probed by Southern blot analysis. The T414G mutation was analyzed by peptide nucleic acid directed clamping PCR. Real-time PCR measured the ratio of mtDNA to nuclear (n)DNA. RESULTS: KC corneas had decreased CO-Iota in areas of corneal thinning. LX-PCR mtDNA digested with restriction enzymes showed expected size bands except for PstI, which showed two additional bands in some KC corneas (2/18). By both LX-PCR (7.4 +/- 3.8 vs. 4.3 +/- 2.7, P < 0.04) and SX-PCR (5.5 +/ 0.55 vs. 2.4 +/- 2.0, P < 0.006), KC corneas had an increased number of smaller sized bands (representing mtDNA deletions/mutations) compared with normal corneas. Southern blot analysis of SX-PCR products confirmed their mtDNA origin. The T414G mutation was not detected in either KC or normal corneas. KC corneas showed a trend of lower mtDNA-to-nDNA ratio (26%, P < 0.7) than did normal corneas. CONCLUSIONS: KC corneas exhibit more mtDNA damage than do normal corneas. The previously reported increased oxidative stress and altered integrity of mtDNA may be related to each other and may be important in KC pathogenesis. PMID- 15790888 TI - Subclinical visual involvement in multiple sclerosis: a study by MRI, VEPs, frequency-doubling perimetry, standard perimetry, and contrast sensitivity. AB - PURPOSE: To evaluate the effectiveness of visual evoked potentials (VEPs), frequency-doubling perimetry (FDP), standard achromatic perimetry (SAP), contrast sensitivity (CS) test, and magnetic resonance imaging (MRI), isolated or in combination, in detecting subclinical impairment of visual function in multiple sclerosis (MS). METHODS: Twenty-two eyes of 11 patients affected by clinically definite MS, without a history of optic neuritis and asymptomatic for visual disturbances, underwent full ophthalmic examination and, in addition, VEPs, FDP, SAP, CS, and MRI. Abnormal results were taken to be as follows: for VEPs, a P100 latency >115 ms; for FDP, abnormal mean deviation (MD) or pattern SD (PSD); for SAP, abnormal MD or PSD; for CS, abnormal CS at one spatial frequency, at least; and for MRI, evidence of at least one demyelinating plaque along the visual pathway. RESULTS: VEPs showed abnormal results in 12 eyes (54.4%), FDP in 11 (50%), SAP in 14 (63.6%), CS in 17 (77.1%), and MRI in 16 (72.7%). In only two (9.1%) eyes of the same patient was no abnormality found. No single test detected all the abnormal eyes. Four (18.2%) eyes had pure optic nerve involvement and the remaining 16 (72.7%) had both pre- and postchiasmal involvement. CONCLUSIONS: In patients affected by clinically definite MS without history of optic neuritis and no visual symptoms, there is a large prevalence of visual pathway involvement that can be diagnosed only by performing multiple tests. The comparison of the tests is also useful to detect the presence of multiple lesions in the same patient. PMID- 15790889 TI - Central corneal thickness correlated with glaucoma damage and rate of progression. AB - PURPOSE: To evaluate whether the amount of glaucomatous optic nerve damage at presentation of the patient and the rate of progression of glaucoma during follow up are related to central corneal thickness. METHODS: The prospective observational clinical study included 861 eyes of 454 white subjects (239 normal eyes of 121 subjects, 250 ocular hypertensive eyes of 118 patients, 372 eyes of 215 patients with chronic open-angle glaucoma). For 567 eyes (304 patients) with ocular hypertension or chronic open-angle glaucoma, follow-up examinations were performed, with a mean follow-up time of 62.7 +/- 33.2 months (median, 60.8; range, 6.2-124.9). All patients underwent qualitative and morphometric evaluation of color stereo optic disc photographs and white-on-white visual field examination. Central corneal thickness was measured by corneal pachymetry. RESULTS: Central corneal thickness correlated significantly (P < 0.001) and positively with the area of the neuroretinal rim and negatively with the loss of visual field. Development or progression of glaucomatous visual field defects detected in 119 (21.0%) eyes was statistically independent of central corneal thickness, in univariate (P = 0.99) and multivariate Cox regression analyses (P = 0.19). CONCLUSIONS: At the time of patient referral, the amount of glaucomatous optic nerve damage correlated significantly with a thin central cornea. Progression of glaucomatous optic nerve neuropathy was independent of central corneal thickness, suggesting that central corneal thickness may not play a major role in the pathogenesis of progressive glaucomatous optic nerve damage. PMID- 15790890 TI - Central corneal thickness and thickness of the lamina cribrosa in human eyes. AB - PURPOSE: Since central corneal thickness may inversely influence the amount and rate of progression of glaucomatous optic nerve damage and because lamina cribrosa thickness may be of importance in susceptibility to glaucoma, it was the purpose of the present study to evaluate whether central corneal thickness is related to lamina cribrosa thickness. METHODS: The histomorphometric study included 111 enucleated nonglaucomatous eyes of 111 white subjects. On anterior posterior histologic sections through the pupil and the central optic disc region, the thickness of the cornea, lamina cribrosa, and peripapillary sclera and the shortest distance between the intraocular space and the cerebrospinal fluid space were measured. Axial length ranged between 20 and 32 mm. RESULTS: Mean central corneal thickness (mean +/- SD: 616.6 +/- 108.3 microm) and mean central lamina cribrosa thickness (378.1 +/- 117.8 microm) were statistically independent of each other (P = 0.15; correlation coefficient, r = 0.14). In a similar manner, lamina cribrosa thickness at the optic disc border was statistically independent of central corneal thickness (P = 0.51; r = 0.06) and peripheral corneal thickness (P = 0.34; r = 0.09). In a parallel way, peripapillary scleral thickness (P = 0.84) and the shortest distance between the prelaminar space and cerebrospinal fluid space (P = 0.10) were statistically independent of central corneal thickness. CONCLUSIONS: In nonglaucomatous human globes, central corneal thickness may not correlate significantly with lamina cribrosa thickness, peripapillary scleral thickness, and shortest distance between intraocular space and cerebrospinal fluid space. Histologic artifact and sectioning methods could partially account for the lack of an association. The study results may suggest clinically that an assumed relationship between central corneal thickness and susceptibility to glaucoma cannot be explained by an anatomic correspondence between corneal thickness and histomorphometry of the optic nerve head. PMID- 15790892 TI - The influence of various substances on the biomechanical behavior of lamina cribrosa and peripapillary sclera. AB - PURPOSE: Changes in the biomechanical properties of the lamina cribrosa (LC) and of the peripapillary sclera (ppSc) may play a role in the pathogenesis of glaucoma. The purpose of this study was to assess the influence of glyceraldehyde, methylglyoxal, and collagenase A on the mechanical properties of the LC and ppSc. METHODS: Two strips of 1-mm width were cut from each of 80 porcine eyes and 24 pairs of enucleated human eyes. One strip contained the LC and the other the adjacent superior ppSc. One half of the strips was divided into groups and treated with 0.5 M glyceraldehyde, 0.5 M methylglyoxal, and 0.1% collagenase A. The other strips served as the control. The stress strain relation was measured in the stress range of 0.02 to 6.0 MPa by a biomaterial tester. RESULTS: Stress values at 20% strain of the human LC changed from 1.97 +/- 1.48 to 3.40 +/- 1.60 MPa after incubation with glyceraldehyde (P = 0.029), from 2.42 +/- 2.22 to 5.46 +/- 1.91 MPa (P = 0.014) after incubation with methylglyoxal, and from 2.43 +/- 1.3 to 1.35 +/- 0.19 MPa after incubation with collagenase A. The stress values of human ppSc without glyceraldehyde were 3.40 +/- 2.59 and 7.45 +/- 4.46 MPa after incubation with glyceraldehyde (P = 0.047), 4.80 +/- 3.05 MPa without methylglyoxal and 16.10 +/- 5.53 MPa (P = 0.001) after incubation with methylglyoxal, 4.14 +/- 2.56 MPa without collagenase A, and 1.97 +/- 0.55 MPa after incubation with collagenase A. At a 20% strain, Young's moduli of the untreated LC were in the range of E = 11.8 to 15.6 MPa and E = 28.5 to 36.0 MPa of the untreated ppSc. CONCLUSIONS: Glyceraldehyde and methylglyoxal increase the stiffness of the LC and of the ppSc in human and in porcine eyes. These substances induce changes in the extracellular matrix according to the Maillard reaction as it occurs during the ageing process or in case of high blood glucose levels. Collagenase reduces the stiffness of the tissues. PMID- 15790893 TI - The effect of age on optic nerve head blood flow. AB - PURPOSE: To examine whether optic nerve head blood flow changes with aging. METHODS: One randomly chosen eye of each of 103 healthy subjects (age range, 22 76 years) was examined by laser Doppler flowmetry. Relative capillary blood flow, velocity, and volume of the moving red blood cells were measured at the temporal and nasal neuroretinal rim of the optic nerve head. For statistical analysis, linear regression analysis and partial correlations were calculated. RESULTS: Velocity increased with age. The Pearson correlation coefficient (R) between age and velocity was 0.49 temporally and 0.56 nasally. The correlation was significantly different from 0 (both P < 0.0005). Volume decreased with increasing age. The Pearson correlation coefficient between age and volume was 0.47 temporally and -0.40 nasally. The correlation was significantly different from 0 (both P < 0.0005). Flow decreased with increasing age. The Pearson correlation coefficient between age and flow was -0.27 temporally and -0.31 nasally. The correlations were significantly different from 0 (P = 0.006 and P = 0.002, respectively). Partial correlation coefficients between the perfusion parameters and age, after correcting for systolic and diastolic blood pressure, heart rate, gender, and IOP, were similar compared to the Pearson correlation coefficients and remained statistically significant. CONCLUSIONS: The perfusion of the optic nerve head is altered with increasing age. The results suggest that the blood supply is reduced in elderly subjects. PMID- 15790891 TI - Comparability of retinal nerve fiber layer thickness measurements of optical coherence tomography instruments. AB - PURPOSE: To compare retinal nerve fiber layer (RNFL) measurements between two ocular coherence tomography (OCT) instruments (OCT 2000 and Stratus OCT; Carl Zeiss Meditec, Dublin, CA) and compare their diagnostic precision. METHODS: One hundred thirty-nine consecutive subjects were imaged (3 x 3.4-mm diameter circular scans) on the same day with each instrument. Thirty-five patients were excluded due to poor-quality images. RNFL thicknesses measured by the two instruments were compared, and receiver operating characteristic (ROC) curves were used to determine diagnostic precision. RESULTS: A randomly selected eye of each of 104 participants (28 with open-angle glaucoma, 40 with suspected glaucoma, and 36 healthy subjects) was analyzed. RNFL thickness measurements generally were thicker with OCT 2000 than with Stratus OCT. The difference in global RNFL thickness between instruments was within 20 microm in 66 (65%) of subjects and within 10 microm (the instrument's limit of resolution) in 25 (25%) subjects. Application of a correction factor to OCT 2000 measurements predicted Stratus OCT RNFL thickness within 10 microm of the observed measurement in 75% of the eyes. For both instruments, highest ROC curve areas (better discrimination between glaucomatous and normal eyes) were found in the inferior sector. Discrimination using global RNFL thickness was better with Stratus OCT than OCT 2000 (P = 0.043). CONCLUSIONS: RNFL thickness measurements measured by OCT 2000 can be approximated to measurements made by Stratus OCT using correction factors calculated by this study. However, there remains considerable variability that exceeds the limits of resolution afforded by the instruments themselves. Therefore comparisons between instruments using these approximations should be interpreted with caution. PMID- 15790895 TI - Molecular analysis of the myocilin gene in Chinese subjects with chronic primary angle closure glaucoma. AB - PURPOSE: Mutations in the myocilin (MYOC) gene have been implicated in juvenile as well as late-onset primary open-angle glaucoma (POAG). Overall, MYOC mutations account for 3% to 5% of cases of POAG worldwide, making it the most significant gene identified so far in glaucoma. Although there are some similarities in the phenotype of POAG and in particular chronic primary angle-closure glaucoma (PACG), little is known about the role of MYOC in the causation of PACG. To address this, the MYOC gene was screened in a cohort of 106 patients with chronic PACG. METHODS: Genomic DNA was extracted from leukocytes of the peripheral blood and exons 1 to 3 of the MYOC gene were PCR amplified and subjected to bidirectional sequencing and analysis. RESULTS: One hundred six patients with chronic PACG of Chinese ethnicity were studied. Sequencing of the MYOC gene in these patients revealed eight sequence variants. Of these, one was a nonsense change, three were missense changes, two were synonymous codon changes, and two were changes in noncoding sequences. These included the Arg46Stop and Thr353Ile mutations, which have been reported in individuals with POAG. However, all the sequence alterations identified have been found in normal Chinese subjects. CONCLUSIONS: The results of this study do not support a role for MYOC mutations in the pathogenesis of chronic PACG in the Chinese. PMID- 15790896 TI - The force required to induce hemivein pulsation is associated with the site of maximum field loss in glaucoma. AB - PURPOSE: To determine the factors associated with central retinal vein pulsation changes in glaucoma and identify any hemiretinal vein pulsation changes and their association with sectoral visual field loss. METHODS: One hundred twenty-six patients with glaucoma and 40 normal subjects had automated perimetry, blood pressure, and intraocular pressure measured. A hemifield sensitivity loss was calculated from the upper and lower halves of each field. Those without spontaneous venous pulsation on the optic disc had an ophthalmodynamometer applied, to measure the minimum ophthalmodynamometric force (ODF) necessary to induce venous pulsation. When ODF was restricted to the hemiveins, the force needed to induce pulsation in each hemivein was measured. RESULTS: Eighty-three patients with glaucoma had no spontaneous venous pulsation. The minimum ODF was strongly correlated with mean deviation (Spearman rank r = -0.475, P < 0.0001). Mixed linear regression analysis showed that mean deviation (P < 0.0001) and pulse blood pressure (P < 0.0001) were significantly associated with minimum ODF. There was a strong association between differences in hemifield sensitivity loss and in hemivein ODF (rank r = 0.369, P < 0.0001, n = 80). Multiple linear regression modeling demonstrated that lower hemivein ODF was independently associated with upper field loss (P = 0.003) and upper hemivein ODF with lower field loss (P < 0.0001). CONCLUSIONS: These venous pulsation findings in glaucoma are independent of blood pressure. The hemifield and hemivein association suggests that the major hemivein change is adjacent to the site of major disc damage. PMID- 15790894 TI - Habituation of retinal ganglion cell activity in response to steady state pattern visual stimuli in normal subjects. AB - PURPOSE: To evaluate autoregulatory changes of retinal ganglion cell (RGC) activity, as measured by the pattern electroretinogram (PERG), when the eye is exposed to a steady state presentation of stimuli that maximize PERG amplitude and blood flow. METHODS: The PERG was recorded from both eyes of 14 normal subjects in response to steady state presentation (4 minutes) of contrast reversing (16.28/s) gratings (1.6 cyc/deg) with different contrast (12%-99%) and mean luminance (40-1.3 cd/m(2)). One temporal period of the stimulus (122.8 ms) was sampled and averaged in packets of 50 sweeps ( approximately 15 seconds each). PERG amplitude and phase were evaluated by Discrete Fourier Transform and displayed as a function of time. Data were fitted with an exponential decay function to evaluate PERG changes with time. RESULTS: For patterns of 99% contrast, the PERG amplitude progressively decreased with time until reaching a plateau approximately 30% lower than the initial amplitude after approximately 2 minutes (habituation). The ratio between initial and plateau amplitude did not change by reducing the stimulus luminance by 1 log unit. However, reducing contrast decreased amplitude habituation. The habituation was abolished at 25% contrast. CONCLUSIONS: Decrease of PERG amplitude with time is consistent with a slow adaptive change of RGC activity in response to high-contrast, steady state stimuli. The authors propose that the initial amplitude represents an index of RGC activity, and the plateau amplitude represents a dynamic equilibrium between RGC activity and the available energy supply. These results are relevant for a better understanding of glaucomatous optic neuropathy. PMID- 15790897 TI - Evaluation of inducible nitric oxide synthase in glaucomatous optic neuropathy and pressure-induced optic nerve damage. AB - PURPOSE: To determine whether inducible nitric oxide synthase (NOS-2) is involved in glaucomatous optic neuropathy. METHODS: Chronic elevation of rat intraocular pressure (IOP) leading to optic nerve damage was induced by episcleral injection of hypertonic saline, which caused sclerosis and blockade of aqueous humor outflow pathways. Expression of NOS-2 in the retina and optic nerve head (ONH) was evaluated by immunohistochemistry, gene array analysis, and quantitative PCR (Q-PCR). Immunohistochemistry was also used to assess the NOS-2 level in the ONH from primary open-angle glaucoma (POAG) and nonglaucomatous human eyes. Finally, an NOS-2 inhibitor, aminoguanidine, administered orally in the drinking water, was tested for its effect on optic nerve injury in rats with ocular hypertension. RESULTS: Chronically elevated IOP in the rat produced optic nerve damage that correlated with pressure change (r(2) = 0.77), but did not increase NOS-2 immunoreactivity in the optic nerve, ONH, or ganglion cell layer. Retinal and ONH NOS-2 mRNA levels did not correlate with either IOP level or severity of optic nerve injury. Similarly, there was no difference in NOS-2 immunoreactivity in the optic nerve or ONH between POAG and nonglaucomatous eyes. Furthermore, aminoguanidine treatment did not affect the development of pressure-induced optic neuropathy in the rat. CONCLUSIONS: As demonstrated by several independent methods, glaucomatous optic neuropathy was not associated with a significant change in the expression of NOS-2 in the retina, ONH, or optic nerve. PMID- 15790898 TI - Relevance vector machine and support vector machine classifier analysis of scanning laser polarimetry retinal nerve fiber layer measurements. AB - PURPOSE: To classify healthy and glaucomatous eyes using relevance vector machine (RVM) and support vector machine (SVM) learning classifiers trained on retinal nerve fiber layer (RNFL) thickness measurements obtained by scanning laser polarimetry (SLP). METHODS: Seventy-two eyes of 72 healthy control subjects (average age = 64.3 +/- 8.8 years, visual field mean deviation = -0.71 +/- 1.2 dB) and 92 eyes of 92 patients with glaucoma (average age = 66.9 +/- 8.9 years, visual field mean deviation = -5.32 +/- 4.0 dB) were imaged with SLP with variable corneal compensation (GDx VCC; Laser Diagnostic Technologies, San Diego, CA). RVM and SVM learning classifiers were trained and tested on SLP-determined RNFL thickness measurements from 14 standard parameters and 64 sectors (approximately 5.6 degrees each) obtained in the circumpapillary area under the instrument-defined measurement ellipse (total 78 parameters). Ten-fold cross validation was used to train and test RVM and SVM classifiers on unique subsets of the full 164-eye data set and areas under the receiver operating characteristic (AUROC) curve for the classification of eyes in the test set were generated. AUROC curve results from RVM and SVM were compared to those for 14 SLP software-generated global and regional RNFL thickness parameters. Also reported was the AUROC curve for the GDx VCC software-generated nerve fiber indicator (NFI). RESULTS: The AUROC curves for RVM and SVM were 0.90 and 0.91, respectively, and increased to 0.93 and 0.94 when the training sets were optimized with sequential forward and backward selection (resulting in reduced dimensional data sets). AUROC curves for optimized RVM and SVM were significantly larger than those for all individual SLP parameters. The AUROC curve for the NFI was 0.87. CONCLUSIONS: Results from RVM and SVM trained on SLP RNFL thickness measurements are similar and provide accurate classification of glaucomatous and healthy eyes. RVM may be preferable to SVM, because it provides a Bayesian derived probability of glaucoma as an output. These results suggest that these machine learning classifiers show good potential for glaucoma diagnosis. PMID- 15790899 TI - Serum deprivation induces apoptotic cell death of transformed rat retinal ganglion cells via mitochondrial signaling pathways. AB - PURPOSE: Apoptosis-related signaling pathways were investigated in a cultured rat retinal ganglion cell (RGC-5) line deprived of growth factors after serum withdrawal from the culture medium. METHODS: RGC-5 cells were subjected to serum deprivation for 2 to 6 days and compared with RGC-5 cells cultured in growth medium containing 10% fetal calf serum. Cell viability was determined by a neutral red dye uptake assay. Apoptosis of RGC-5 cells was established by DNA laddering. The expression of various apoptosis-related genes was investigated by immunoblot analysis, and or reverse transcription polymerase chain reaction (RT PCR) analysis. The redox state of the cell was determined by biochemical methods, including NF-kappaB binding activity by electrophoretic mobility gel shift assays (EMSA) and mitochondrial damage by JC-1 (5,5', 6,6'-tetrachloro 1,1',3,3' tetraethylbenzimidazolyl-carbocyanine iodide) staining, using live cell confocal microscopy and cytosolic release of cytochrome c. RESULTS: Fifty percent cell loss was evident after 2 days of serum deprivation, as demonstrated by neutral red dye uptake assay. This cell loss was due to apoptotic cell death, as established by DNA laddering. The oxidative state of serum-deprived RGC-5 cells was perturbed as suggested by the increase in malonyldialdehyde (MDA) and a decrease in reduced glutathione (GSH) levels in cell lysates. The apoptosis of the RGC-5 cells was associated with the activation of caspase-3, -8, and -9, and increased levels of Bax with corresponding decreases in Bcl-2 levels and NF kappaB (NF-kappaB) binding activity. Serum deprivation was also associated with a loss of mitochondrial function, as revealed by cytosolic release of cytochrome c and JC-1 staining of mitochondria of dying RGC-5 cells. CONCLUSIONS: Taken together, these results indicate that serum withdrawal induces apoptotic cell death in RGC-5 cells via mitochondrial pathways. These studies lead to the speculation that growth factor deprivation arising from blockade of retrograde transport of neurotrophins may involve similar mechanism(s) of retinal ganglion cell death in glaucoma. PMID- 15790900 TI - Effects of the NF-kappaB inhibitor pyrrolidine dithiocarbamate on experimentally induced autoimmune anterior uveitis. AB - PURPOSE: To determine the effect of pyrrolidine dithiocarbamate (PDTC), a nuclear factor (NF)-kappaB inhibitor, on chemokine and chemokine receptor expression and thus elucidate the role of NF-kappaB in the pathogenesis of experimental autoimmune anterior uveitis (EAAU). METHODS: Uveitis was induced in Lewis rats with the injection of melanin-associated antigen into the footpad. PDTC (200 mg/kg and 100 mg/kg) was administered intraperitoneally daily, beginning 1 day after the immunization. The clinical inflammatory activity of the anterior chamber was recorded daily and scored. Immunohistochemical staining and an electrophoretic mobility shift assay assessed the effect of PDTC on NF-kappaB activation in the iris/ciliary body tissues. Gene expression profiles of chemokine and chemokine receptors were semiquantitatively examined by reverse transcriptase-polymerase chain reaction (RT-PCR). Aqueous chemokine levels were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: PDTC significantly attenuated the clinical scores and monocyte/lymphocyte infiltration in rats with EAAU. PDTC effectively inhibited NF-kappaB activation in the iris and ciliary body, and markedly inhibited the expression of chemokine genes, including monocyte chemoattractant protein (MCP)-1, regulated-on-activation normal T-cell expressed and secreted (RANTES), and interleukin (IL)-8 and chemokine receptors genes including CCR2, CCR5, and CXCR3. CONCLUSIONS: Activation of NF-kappaB appears to play an important role in the pathogenesis of EAAU, through transcriptional control of MCP-1, RANTES, and IL-8 gene expression. Blocking NF-kappaB reduces ocular inflammation and may be an effective strategy in the treatment of acute anterior uveitis. PMID- 15790901 TI - Internalization of Pseudomonas aeruginosa is mediated by lipid rafts in contact lens-wearing rabbit and cultured human corneal epithelial cells. AB - PURPOSE: The internalization of Pseudomonas aeruginosa (PA) in nasal and tracheal epithelium has recently been shown to involve the formation of cholesterol- and sphingolipid-rich plasma membrane domains (lipid rafts). The purpose of this study was to investigate the role of lipid rafts in PA internalization by corneal epithelium in vivo, in vitro, and after contact lens wear. METHODS: Lipid raft formation was evaluated in rabbit corneas with and without contact lens wear and a human corneal epithelial (hTCEpi) cell line before and after PA infection with cornea-pathogenic strains by staining with FITC-conjugated cholera toxin beta subunit, known to bind the lipid raft component GM1. Bacterial internalization was assessed by gentamicin survival assay. The role of lipid rafts in PA internalization was evaluated by pretreatment of hTCEpi cells with cholesterol metabolism inhibitors. The interaction of PA with lipid rafts was confirmed by flow cytometry. RESULTS: Contact lens wear in rabbits induced lipid raft formation in occasional surface corneal epithelial cells. Subsequent PA exposure showed preferential binding to lipid raft-forming cells, leading to lipid raft aggregation and PA internalization. A similar sequence of lipid raft formation and PA internalization was also observed in hTCEpi for all PA strains. Internalization of all PA strains was blocked by three cholesterol metabolism inhibitors (P < 0.01). Flow cytometry showed an association of PA with rafts. CONCLUSIONS: These findings demonstrate that contact-lens-mediated PA internalization involves lipid raft formation. Also, hTCEpi cells may be used as an experimental model for studying further the molecular mechanism(s) of PA infection in the corneal epithelium. PMID- 15790902 TI - Cytokine profiles and inflammatory cells during HSV-1-induced acute retinal necrosis. AB - PURPOSE: To investigate infiltrating cells, cytokines, and kinetics of cytokine expression during acute retinal necrosis (ARN) in the uninoculated eye after inoculation of herpes simplex virus (HSV)-1 into the anterior chamber of one eye of BALB/c mice. METHODS: At different time points after inoculation of 2 x 10(4) plaque-forming units (PFU) HSV-1 (KOS strain) or an equivalent volume of Vero cell extract in cell culture medium, the uninoculated eyes were enucleated. RT PCRs for TNFalpha, IFNgamma, and IL-4 and immunohistochemical staining were performed to identify infiltrating cells and cytokines. Cytometric bead array was used to measure the levels of TNFalpha, IFNgamma, and IL-4 protein. RESULTS: CD4(+) T cells, F4/80(+) macrophages, Gr-1(+) polymorphonuclear cells (PMNs), and CD19(+) B cells were detected in the uninoculated eye of virus-infected mice. Furthermore, RPE65(+) retinal pigment epithelial (RPE) cells and activated Muller cells were also detected in the ARN lesion. TNFalpha, IFNgamma, and IL-4 mRNA and protein were upregulated during the evolution of ARN in HSV-1-infected contralateral eyes compared with levels in control subjects. Immunohistochemistry revealed that cytokines were produced by infiltrating cells as well as by resident retinal cells. CONCLUSIONS: The results of these studies support the idea that T cells and cytokines are actively involved in HSV-1 retinitis. They also suggest that PMNs, B cells, and/or macrophages, as well as resident retinal cells, such as RPE and activated Muller cells, also play a role in the pathogenesis of HSV-1 retinitis. PMID- 15790904 TI - Pathogenesis of Staphylococcus in the rabbit anterior chamber. AB - PURPOSE: To investigate the host defense against Staphylococcus in the rabbit anterior chamber. METHODS: The bactericidal activity of rabbit aqueous humor was investigated in vitro. Rabbit anterior chambers were injected with viable Staphylococcus aureus or Staphylococcus epidermidis (1,000 or 500,000 colony forming units [CFU]), killed bacteria, culture supernatants of either organism, or purified S. aureus alpha-toxin. CFU as well as phospholipase (PLA(2)) and myeloperoxidase (MPO) activities of aqueous humor were determined up to 25 hours postinfection (PI). RESULTS: The number of viable S. aureus or S. epidermidis was significantly reduced when incubated with aqueous humor for 30 minutes (P 0.01) reduction in the diameter of the lesion, a nonsignificant increase in VA and SLO-CM, and a still significant increase in FERG and PERG amplitudes. CONCLUSIONS: In myopic CNV eyes, PDT induces an increase, though not significant, in VA and macular sensitivity. These changes may be related to a reduction in the diameter of the lesion, with an improvement in the function of both ganglionic and preganglionic elements of the macular region, as suggested by the improvement in FERG and PERG responses. PMID- 15790916 TI - The interaction of indocyanine green with human retinal pigment epithelium. AB - PURPOSE: This study sought to examine the in vitro interaction of human RPE cells with indocyanine green (ICG). The interaction between ICG and the RPE may have clinical relevance in the interpretation of clinical ICG angiography. METHOD: Cultured primary human RPE cells were incubated with ICG. Infrared fluorescence microscopy was used to detect RPE cell ICG fluorescence. The proportions of cells exhibiting ICG infrared fluorescence were quantified. Separate RPE cell populations were incubated with ouabain for 24 and 72 hours, respectively, before addition of the ICG to examine its effect on the uptake of ICG. The effect of ouabain on cell viability was assessed with trypan blue exclusion. RESULTS: Normal human RPE cells incubated with ICG exhibited strong infrared fluorescence. Exposure to ouabain for 24 hours before incubation with ICG had little effect on cell viability but significantly reduced cellular ICG fluorescence. In contrast, exposure to ouabain for 72 hours reduced cell viability and increased cellular ICG fluorescence. CONCLUSIONS: Cultured human RPE cells take up ICG dye. ICG uptake by RPE cells may involve active transport, as cells incubated with ouabain for 24 hours showed no reduction in cell viability but exhibited reduced infrared fluorescence. The paradoxical increased uptake of ICG into the cells after more prolonged exposure to ouabain may be due to ICG's movement through the damaged cell membrane. Fluorescence due to ICG uptake by RPE has clinical relevance in that it contributes to the fluorescence patterns observed in ICG angiography. PMID- 15790917 TI - Histology of the vitreoretinal interface after indocyanine green staining of the ILM, with illumination using a halogen and xenon light source. AB - PURPOSE: To assess the histology of the retinal surface after staining of the inner limiting membrane (ILM) with indocyanine green (ICG) followed by illumination with halogen or xenon light sources in human donor and porcine eyes. METHODS: Ten eyes of six human donors and six porcine eyes were used in the study. In human donor eyes, the postmortem time varied between 7 and 38 hours, and porcine eyes were evaluated 9 hours after death. In all eyes, the vitreous was removed, and a few drops of 0.5% ICG were poured over the trephined posterior pole and carefully washed out after a period of 1 minute, with balanced salt solution. Then the stained retina was illuminated for 3 minutes with different light sources: a halogen light source of 145-W power or a xenon light source of 50-W power. Adjacent, unstained retina of each eye served as a control to assess postmortem artifacts. In two human and two porcine eyes ICG was applied without illumination. Retinal specimens were evaluated by light and electron microscopy. RESULTS: In human eyes, severe disorganization of the innermost retina and ILM loss were observed after ICG application with subsequent illumination with the halogen light source. After illumination with the xenon light source, there was only slight vacuolization of the innermost retina, with mostly intact Muller cells. The ILM remained in situ in relation to the retinal surface. Intact cellular architecture was found in all specimens after ICG staining without subsequent illumination and control specimens of unstained retina. In porcine eyes, no impact attributable to the light source or ICG alone was noted in this experimental setting. CONCLUSIONS: These findings suggest that adverse effects of ICG at the retinal surface may depend on the light source used during vitrectomy and correlate with the emission spectrum of the different light sources. In addition, care should be taken when comparing results obtained in human eyes and porcine eyes. PMID- 15790918 TI - Downregulation of cone-specific gene expression and degeneration of cone photoreceptors in the Rpe65-/- mouse at early ages. AB - PURPOSE: RPE65 is essential for the generation of 11-cis retinal. Rod photoreceptors in the RPE65-knockout (Rpe65(-/-)) mouse are known to degenerate slowly with age. This study was designed to examine cone photoreceptors and the expression of cone-specific genes in the Rpe65(-/-) mouse. METHODS: Gene expression changes were identified by microarray and confirmed by real-time RT PCR. Cone photoreceptors were stained by peanut agglutinin (PNA) lectin in the flatmounted retina. The 9- or 11-cis retinal was supplied by intraperitoneal injections. RESULTS: The short-wavelength (SWL) cone opsin mRNA was markedly decreased at 2 weeks of age, whereas the decrease in the middle-wavelength (MWL) cone opsin mRNA occurred relatively later in age. In contrast, the rhodopsin mRNA level did not show any significant change at all the ages analyzed. Consistent with the cone opsin changes, the cone transducin alpha-subunit mRNA decreased at both 4 and 8 weeks of age, whereas again the rod transducin alpha-subunit did not show any significant change. Rpe65(-/-) mice showed significant cone loss in both the central and ventral retina between 2 and 3 weeks of age. Administration of 9- or 11-cis retinal to Rpe65(-/-) mice 2 weeks of age increased cone density by twofold in these areas. CONCLUSIONS: In the Rpe65(-/-) mouse, the expression of cone-specific genes is downregulated and is accompanied by cone degeneration at early ages. Early administration of 9- or 11-cis retinal can partially prevent cone loss, suggesting that the absence of 11-cis chromophore may be responsible for the early cone degeneration. PMID- 15790919 TI - Cone and rod dysfunction in fundus albipunctatus with RDH5 mutation: an electrophysiological study. AB - PURPOSE: A prior study showed that some patients with fundus albipunctatus (FA) have severely reduced full-field cone ERGs. The purpose of this study was to investigate the frequency of cone dysfunction and to determine the cause of the reduced full-field cone ERGs in patients with FA and whether the rod system is affected in patients with FA. METHODS: Sixteen consecutive patients with FA (from 1993 to 2003; eight males, eight females; mean age, 25.4 years) with an RDH5 gene mutation were studied. The amplitudes and implicit times of the standard cone ERGs in the patients with FA were compared to those obtained from normal subjects (n = 55). The a-waves of cone ERGs were also elicited by a bright flash and were fitted to a mathematical model of the a-wave. Rod ERG responses were elicited by dim blue flashes after 3 hours of dark adaptation. RESULTS: The amplitude of the b-wave of the cone ERG in the FA group varied considerably from within the normal limits to markedly decreased. Six of 16 patients with FA had b-wave amplitudes that were smaller than the lowest limit of the control subjects. The degree of cone dysfunction tended to be more severe in older patients. The analysis of the cone a-wave demonstrated that R(m) (maximal response amplitude) in the patients with FA with reduced standard cone ERGs was significantly smaller than that in control subjects. Rod ERGs were also reduced in the patients with FA who had reduced cone ERGs. CONCLUSIONS: In patients with FA, 38% had extensive cone dysfunction. The reduced full-field cone ERGs were mainly due to the loss of cone photoreceptors, and the rod system was also affected in some patients. PMID- 15790920 TI - Thresholds for activation of rabbit retinal ganglion cells with relatively large, extracellular microelectrodes. AB - PURPOSE: To investigate the responses of retinal ganglion cells (RGCs) to electrical stimulation, using electrodes comparable in size to those used in human studies investigating the feasibility of an electronic retinal prosthesis. METHODS: Rabbit retinas were stimulated in vitro with current pulses applied to the inner surface with 125- and 500-mum diameter electrodes while the responses of RGCs were recorded extracellularly. RESULTS: Both short-latency (SL; 3-5 ms) and long-latency (LL; >/=9 ms) responses were observed after electrical stimulation within the receptive field of an RGC. With short, 0.1-ms current pulses, the threshold current for the SL cell response was significantly lower than that for the LL cell response. With long (10- to 20-ms) pulses, the threshold currents for the SL and LL cell responses were very similar. The threshold current for the SL cell response increased more steeply than did the LL cell response when the electrode was displaced from the point of lowest electrical threshold, either above or along the surface of the retina. Stimulation of an RGC axon outside of the cell's receptive field produced only an SL response. For 0.1-ms duration pulses, the threshold current for the axonal response was significantly higher than the threshold current for the SL cell response. At pulse durations > 1 ms, the thresholds were very similar. CONCLUSIONS: RGC responses to electrical stimulation depend on the current pulse duration and location of the stimulating electrode. For an epiretinal prosthesis, short-duration current pulses may be preferable since they result in a more localized activation of the retina. PMID- 15790921 TI - CNTF and BDNF have similar effects on retinal ganglion cell survival but differential effects on nitric oxide synthase expression soon after optic nerve injury. AB - PURPOSE: To investigate the effect of ciliary neurotrophic factor (CNTF) and brain-derived neurotrophic factor (BDNF) on retinal ganglion cell (RGC) survival and nitric oxide synthase (NOS) expression in the retina during the early phase of optic nerve (ON) injury, and to examine whether intraperitoneal application of the NOS scavenger nitro-l-arginine (l-NA) could protect the injured RGCs. METHODS: RGCs were retrogradely labeled with granular blue 3 days before the ON was intraorbitally transected. RGC survival was examined 1 week after ON transection and intraocular injection of CNTF and/or BDNF, or 1 to 2 weeks after daily intraperitoneal injection of the NOS inhibitor l-NA. NOS expression was examined by NADPH-diaphorase histochemistry and neuronal NOS (nNOS) immunohistochemistry, and nNOS-positive cells were identified by various staining approaches. RESULTS: Both CNTF and BDNF significantly increased RGC survival 1 week after ON injury. In the ganglion cell layer (GCL), CNTF did not increase the number of NADPH-diaphorase positive ((+)) cells but appeared to reduce the intensity of NADPH-diaphorase staining, whereas BDNF increased the number of NADPH-diaphorase(+) cells and also appeared to enhance the intensity of NADPH diaphorase staining. In the GCL, amacrine cells but not RGCs were nNOS(+). Some macrophages were also nNOS(+). In contrast, no amacrine cells were nNOS(+) in the inner nuclear layer. Daily intraperitoneal injection of l-NA at appropriate concentrations promoted RGC survival for 1 or 2 weeks after ON injury. CONCLUSIONS: Both CNTF and BDNF protected RGCs after ON injury. CNTF and BDNF acted differently on NOS expression in the GCL. Intraperitoneal injections of l NA at appropriate dosages enhance RGC survival. PMID- 15790922 TI - Tissue type plasminogen activator facilitates NMDA-receptor-mediated retinal apoptosis through an independent fibrinolytic cascade. AB - PURPOSE: To investigate the association between apoptosis and the fibrinolytic system in retinal cell damage. METHODS: Tissue type plasminogen activator deficient (tPA(-/-)), urokinase type plasminogen activator-deficient (uPA(-/-)), plasminogen activator inhibitor-1-deficient (PAI-1(-/-)), alpha2 antiplasmin deficient (alpha2 AP(-/-)) mice, and their wild-type counterparts were used. Retinal cell damage was induced by intravitreal injection of the excitotoxin N methyl-d-aspartate (NMDA). The TdT-dUTP terminal nick-end labeling (TUNEL) method was used to examine retinal cell damage. RESULTS: tPA(-/-) mice were resistant to retinal cell damage caused by administration of NMDA, and PAI-1(-/-) mice were more injured than their wild-type. No significant difference was observed between uPA(-/-) or alpha2 AP(-/-) and their wild-type mice. CONCLUSIONS: The results strongly suggest that endogenous tPA, but not uPA acts as a facilitator in NMDA induced retinal cell damage, and that its mechanism may not be associated with cleavage of plasminogen into plasmin in the fibrinolytic cascade. PMID- 15790923 TI - Altered gene expression of angiogenic factors induced by calcium-mediated dissociation of retinal pigment epithelial cells. AB - PURPOSE: To examine the effect of loss of cell-cell contacts on the gene expression of vascular endothelial growth factor (VEGF) and other factors in primary culture of human retinal pigment epithelial (RPE) cells with real-time reverse transcription-PCR. METHODS: The dissociation of postconfluent RPE cells was induced by calcium chelation, low-calcium medium, anti-E-cadherin, and anti-N cadherin antibodies. Total RNA was isolated from the cultured RPE cells and reverse transcribed to cDNA. VEGF was quantified by real-time PCR with a fluorescence detector. VEGF isoforms were differentially measured by specific exon-spanning primers. Besides VEGF, the gene expression levels of some other growth factors were also examined in calcium-mediated dissociation. RESULTS: Disruption of cell-cell contacts of RPE cells was induced by calcium chelation and low-calcium medium, but not by anti-E-cadherin and anti-N-cadherin antibodies. Calcium-mediated dissociation of RPE cells significantly increased the gene expression levels of VEGF. The mRNA levels of VEGF increased by 6.3-fold on treatment with EGTA and by 4.7-fold in the low-calcium medium at 6 hours. Splice variants of VEGF showed the differential pattern of gene expression. Whereas the expression of VEGF(121) and VEGF(165) was upregulated on calcium induced dissociation of RPE cells, that of VEGF(145) and VEGF(189) was unchanged. VEGF(206) was not detected. On calcium-induced dissociation, bFGF, IL-6, matrix metalloproteinase (MMP)-1, and placental growth factor (PlGF) were upregulated, whereas acidic (a)FGF and pigment epithelium-derived factor (PEDF) were both downregulated. CONCLUSIONS: The results show that loss of intercellular contacts promotes increased gene expression of VEGF and other angiogenic factors in human RPE cells. PMID- 15790924 TI - Impaired opsin targeting and cone photoreceptor migration in the retina of mice lacking the cyclic nucleotide-gated channel CNGA3. AB - PURPOSE: To characterize molecular and cellular changes in the mouse retina caused by the genetic deletion of the cone cyclic nucleotide-gated channel (CNG) subunit CNGA3. METHODS: Retinas of wild-type and CNGA3-deficient (CNGA3(-/-)) mice from 9 days up to 22 months of age were analyzed by immunohistochemistry, electron microscopy, and molecular biological methods. RESULTS: CNGA3(-/-) cones failed to transport opsins into outer segments, downregulated various proteins of the phototransduction cascade, and induced apoptotic death. Loss of CNGA3 did not affect the transcription of cone-specific genes. Cone degeneration was evident from the second postnatal week on, and it proceeded significantly faster in the ventral than in the dorsal part of the retina. Ventral cones were completely missing after the third postnatal month, whereas residual dorsal cones were present, even in 22-month-old knockout mice. CNGA3(-/-) cone somata exhibited profoundly delayed migration during postnatal development. At the time of eye opening, most CNGA3(-/-) cones had displaced somata localized close to or in the outer plexiform layer. These cones lacked the characteristic synaptic pedicle, but revealed synapselike contacts to second-order neurons at their somata. At later stages, most of the surviving CNGA3(-/-) cones had correctly located somata and morphologically normal synapses. CONCLUSIONS: The loss of CNGA3 impairs the targeting of cone opsins and the expression of other visual cascade proteins. In addition, CNGA3 appears to be essential for normal postnatal migration of cone somata. After loss of cone outer segment proteins, CNGA3(-/-) cones induce apoptotic cell death. PMID- 15790925 TI - Resensitization of P2Y receptors by growth factor-mediated activation of the phosphatidylinositol-3 kinase in retinal glial cells. AB - PURPOSE: To determine whether activation of receptor tyrosine kinases enhances the responsiveness of purinergic P2Y receptors in Muller glial cells, known to induce Muller cell proliferation. METHODS: The P2Y receptors of primary cultured Muller cells of the guinea pig were desensitized by short (30 seconds to 10 minutes)- and long (24 or 48 hours)-term application of adenosine 5'-triphosphate (ATP). Recordings of ATP-evoked intracellular calcium responses showed whether short-term application of different growth factors evoke a resensitization of the receptors. Coapplication of pharmacologic inhibitors showed whether activation of protein kinases is involved in receptor resensitization. RESULTS: Both short- and long-term incubation with ATP induced a significant P2Y receptor desensitization, which was indicated by a strongly reduced intracellular calcium mobilization and which lasted for at least 48 hours. However, the receptors were significantly resensitized after short-term application of platelet-derived, epidermal, or nerve growth factors. The growth factor-mediated resensitization was dependent on an intact cytoskeleton and on the activation of protein phosphatases and of the phosphatidylinositol-3 kinase (PI3K), but was independent of the activation of protein kinase C, src kinases, or extracellular signal-regulated kinases. CONCLUSIONS: The results show that activation of receptor tyrosine kinases causes, via activation of PI3K and protein phosphatases, a resensitization of P2Y receptors formerly desensitized by agonist application. The growth factor mediated resensitization may underlie the previously observed enhanced responsiveness of P2Y receptors in retinal glial cells in experimental retinal detachment and proliferative vitreoretinopathy and may contribute to the induction of reactive gliosis and Muller cell proliferation. PMID- 15790926 TI - Retinal correspondence of monocular receptive fields in disparity-sensitive complex cells from area V1 in the awake monkey. AB - PURPOSE: To explore the neural mechanisms underlying disparity sensitivity in complex cells of the macaque visual cortex, the relationship between interocular receptive field (RF) positional shift and disparity sensitivity was studied in area V1. METHODS: Single-unit recordings were made from area V1 of awake Macaca mulatta. Monocular RFs were mapped by means of a reverse cross-correlation technique, and their centers were determined after performing a bidimensional Gaussian function fitting. Interocular RF shifts were calculated for both bright and dark stimuli. Similarly, Gabor adjustments were obtained from disparity profiles to bright and dark dynamic random-dot stereograms (RDSs). RESULTS: Twenty-five complex cells were studied. The response profiles to disparity were similar for bright and dark RDSs. Interocular RF positional shift correlated significantly with both the peaks of Gabor fittings of disparity-sensitivity profiles and the peaks of the Gaussian envelopes of these Gabor fittings. Correlation between interocular RF positional shift and the peaks of the Gaussian envelopes was stronger than correlation between interocular RF positional shift and peaks of Gabor fittings. CONCLUSIONS: Interocular shift of monocular RFs is more related to the center of the range of disparities to which the cell is sensitive, than to the preferred disparity of the cell. PMID- 15790928 TI - Oxidative enzymopathies and vascular disease. AB - In the vasculature, reactive oxygen species (ROS) generated by both mitochondrial respiration and enzymatic sources serve as integral components of cellular signaling and homeostatic mechanisms. Because ROS are highly reactive biomolecules, the cellular redox milieu is carefully maintained by small-molecule antioxidants and antioxidant enzymes to prevent the deleterious consequences of ROS excess. When this redox balance is perturbed, because of either increased ROS production or decreased antioxidant capacity, oxidant stress is increased in the vessel wall and, if not offset, vascular dysfunction ensues. A number of heritable polymorphisms of pro-oxidant enzymes, including 5-lipoxygenase, cyclooxygenase-2, nitric oxide synthase-3, and NAD(P)H oxidase, have been identified and found to modulate ROS production and, thereby, the risk of atherothrombotic cardiovascular disease in individuals with these genetic polymorphisms. Similarly, heritable deficiency of the antioxidant enzymes catalase, glutathione peroxidases, glutathione-S-transferases, heme oxygenase, and glucose-6-phosphate dehydrogenase favors ROS accumulation, and has been associated with an increased risk of vascular disease. Individually, each of these polymorphisms imposes a state of uncompensated oxidant stress on the vasculature and collectively comprise the oxidative enzymopathies. PMID- 15790927 TI - Characteristics of the normative database for the Humphrey matrix perimeter. AB - PURPOSE: The Humphrey Matrix (Carl Zeiss Meditec, Dublin CA; Welch-Allyn, Skaneateles, NY) is a high-spatial-resolution perimeter that uses frequency doubling stimuli. It incorporates an efficient test strategy that assumes that age, eccentricity, and test procedure type have only small effects on sensitivity. The results used to create the normative database for the perimeter were examined, to see whether these assumptions were met and to examine the form of the normative data. METHOD: Visual fields were measured (Matrix 30-2, 24-2, 10 2 and Macula patterns) in >275 subjects judged to be normal by a battery of clinical procedures. The right eye was always tested first. RESULTS: Sensitivity decreased by approximately 0.7 dB per age decade across all eccentricities; sensitivity decreased with eccentricity, typically by <5 dB at the most peripheral points tested. Although there was no systematic difference in sensitivity between the 30-2 and 24-2 tests, the Macula test sensitivities were typically 1 dB higher than for the 10-2 test. Sensitivity in the left eye was slightly lower than in the right, with the difference being significantly greater in the temporal visual field. In most test locations, the 95% confidence interval of normal sensitivity was approximately 6 dB below the median sensitivity. CONCLUSIONS: The performance of the test strategy in the Matrix perimeter is appropriately matched to the response characteristics of the normal population. The finding of a spatially nonuniform difference in sensitivity between left and right eyes is attributable to light-adaptation differences between the eyes. This effect is accounted for in the perimeter's normative database. PMID- 15790929 TI - Analysis of Gas6 in human platelets and plasma. AB - OBJECTIVE: Gas6 is a member of the vitamin K-dependent protein family. Gas6 deficient mice were found to be resistant to thrombosis because of defective platelet function. Mouse Gas6 was demonstrated to be present in platelets and found to be involved in platelet aggregation. The aim of this study was to investigate the presence of Gas6 in human platelets and plasma and determine its role in platelet function. METHODS AND RESULTS: The presence of Gas6 in human platelets and plasma was analyzed using sensitive immunologic methods. Mass spectrometry and ELISA were used to identify and quantify Gas6 in plasma. Gas6 was demonstrated to be present in human plasma, at a concentration determined to be 13 to 23 ng/mL (0.16 to 0.28 nM). Furthermore, plasma Gas6 levels were found to be lower in patients administered with warfarin. However, Gas6 was undetectable in human platelets. CONCLUSIONS: This is the first report to identify and quantify Gas6 in human plasma. However, Gas6 protein was not detected in human platelets, suggesting that any potential platelet-specific function could be because of Gas6 from the circulation. These findings open up new directions regarding the role of Gas6 in normal and pathophysiological situations such as inflammation, autoimmune disease, thrombosis and arteriosclerosis. PMID- 15790930 TI - Fenofibric acid, an active form of fenofibrate, increases apolipoprotein A-I mediated high-density lipoprotein biogenesis by enhancing transcription of ATP binding cassette transporter A1 gene in a liver X receptor-dependent manner. AB - OBJECTIVE: Fibrates are widely used drugs to reduce plasma triglyceride and increase high-density lipoprotein. Their active forms, fibric acids, are peroxisome proliferator-activated receptor-alpha activators, but no direct evidence has been demonstrated for their activation of ATP-binding cassette transporter A1 (ABCA1) in relation to clinically used fibrates. We investigated the reaction of fenofibric acid in this regard. METHODS AND RESULTS: Fenofibric acid was examined for the effect of increase of ABCA1 activity. It enhanced ABCA1 gene transcription and its protein level in macrophage cell line cells and fibroblasts and increased apolipoprotein A-I-mediated cellular lipid release, all in a dose-dependent manner. Enhancement of the gene transcription was examined by using a reporter assay system for liver X receptor responsive element (LXRE) and its inactive mutant. The results demonstrated that the effect of fenofibric acid is dependent on active LXRE. CONCLUSIONS: Fenofibric acid increased transcription of ABCA1 gene in a liver X receptor-dependent manner. PMID- 15790931 TI - Elevated interleukin-18 levels are associated with the metabolic syndrome independent of obesity and insulin resistance. AB - OBJECTIVE: Activated innate immunity is thought to be involved in the pathogenesis of metabolic syndrome and type 2 diabetes. Interleukin-18 (IL-18) is a pleiotropic proinflammatory cytokine with important regulatory functions in the innate immune response. We sought to determine whether an elevated IL-18 concentration was a risk predictor for metabolic syndrome in a community population independent of obesity and hyperinsulinemia. METHODS AND RESULTS: A representative general population, aged 27 to 77 years, without clinical diabetes was studied for clinical and biochemical risk factors for metabolic syndrome. Serum IL-18 concentration measured in 955 subjects correlated with metabolic syndrome traits including body mass index (BMI), waist circumference, triglyceride, high-density lipoprotein (inversely), and fasting glucose and insulin levels (all P<0.001). Mean IL-18 levels rose progressively with the increasing number of metabolic risk factors (ANOVA P<0.001). After adjusting for age, gender, BMI, and insulin levels, increasing IL-18 tertiles were associated with an odds ratio for metabolic syndrome of 1.0, 1.42, and 2.28, respectively (P trend=0.007). The graded risk relation was even stronger in nonobese subjects and not attenuated when adjusted for C-reactive protein and IL-6 levels. CONCLUSIONS: Our findings support the hypothesis that activation of IL-18 is involved in the pathogenesis of the metabolic syndrome. PMID- 15790933 TI - Transcriptional regulation of apolipoprotein A5 gene expression by the nuclear receptor RORalpha. AB - OBJECTIVE: The newly identified apolipoprotein A5 (APOA5), selectively expressed in the liver, is a crucial determinant of plasma triglyceride levels. Because elevated plasma triglyceride concentrations constitute an independent risk factor for cardiovascular diseases, it is important to understand how the expression of this gene is regulated. In the present study, we identified the retinoic acid receptor-related orphan receptor-alpha (RORalpha) as a regulator of human APOA5 gene expression. METHODS AND RESULTS: Using electromobility shift assays, we first demonstrated that RORalpha1 and RORalpha4 proteins can bind specifically to a direct repeat 1 site present at the position -272/-260 in the APOA5 gene promoter. In addition, using transient cotransfection experiments in HepG2 and HuH7 cells, we demonstrated that both RORalpha1 and RORalpha4 strongly increase APOA5 promoter transcriptional activity in a dose-dependent manner. Finally, adenoviral overexpression of hRORalpha in HepG2 cells led to enhanced hAPOA5 mRNA accumulation. We show that the homologous region in mouse apoa5 promoter is not functional. Moreover, we show that in staggerer mice, apoa5 gene is not affected by RORalpha. CONCLUSIONS: These findings identify RORalpha1 and RORalpha4 as transcriptional activators of human APOA5 gene expression. These data suggest an additional important physiological role for RORalpha in the regulation of genes involved in lipid homeostasis and probably in the development of atherosclerosis. PMID- 15790932 TI - Polymorphonuclear leukocytes may impair endothelial function: results of crossover randomized study of lipid-lowering therapies. AB - OBJECTIVES: To examine whether polymorphonuclear leukocytes (PMNs) in hypercholesterolemia (HC) are activated to generate large amount of superoxide in vivo and hence impair endothelial function and, if so, whether statins, which possess anti-inflammatory properties, may restore PMN-mediated endothelial dysfunction. METHODS AND RESULTS: At baseline, subjects with HC showed impaired endothelial function (P<0.001), estimated by flow-mediated vasodilation of the brachial artery, and increased susceptibility of low-density lipoprotein (LDL) to oxidation (P<0.0001) compared with control subjects. PMNs obtained from HC produced greater amount of superoxide (P<0.0001), showed higher adhesiveness to cultured endothelial cells (HUVECs) (P<0.0001), and impaired endothelial nitric oxide synthase (eNOS) Ser1177 phosphorylation of HUVECs compared with controls (P<0.001). Crossover administration of fluvastatin or colestimide for 3 months lowered LDL to the same levels (P<0.001 for both). Endothelial function was restored (P<0.0001). LDL oxidation (P<0.0001) and superoxide release from PMNs (P<0.0001) were diminished only in fluvastatin but not in colestimide arm. Fluvastatin attenuated PMN adhesion to HUVECs (P<0.0001) and restored eNOS Ser1177 phosphorylation of HUVECs (P<0.001). CONCLUSIONS: Statins may improve endothelial function at least in part by inactivating neutrophils independently of LDL reduction. Our results raise a novel concept that polymorphonuclear leukocytes may attack endothelia and play a pivotal role in the pathogenesis of atherosclerosis. PMID- 15790934 TI - Statins reduce interleukin-6-induced C-reactive protein in human hepatocytes: new evidence for direct antiinflammatory effects of statins. AB - OBJECTIVE: Besides its predictive role in determining cardiovascular risk, C reactive protein (CRP) may exert direct proatherogenic effects through proinflammatory properties. CRP is mainly produced by hepatocytes in response to interleukin-6 (IL-6) and is then released into the systemic circulation. 3 hydroxy-3-methylglutaryl (HMG)-coenzyme A (CoA) reductase inhibitors, or statins, significantly reduce cardiovascular events and mortality in patients with or without coronary artery disease and reduce plasma CRP levels in humans. However, the mechanism by which statins reduce plasma CRP levels remains unknown. METHODS AND RESULTS: In this study, we report that statins limit both protein and RNA levels of IL-6-induced CRP in human hepatocytes. These effects are reversed by l mevalonate and mimicked by an inhibitor of the geranylgeranyltransferase. IL-6 induced CRP production requires the binding of IL-6 to its cognate receptors, which results in activation and phosphorylation of the transcription factor STAT3. We provide evidence that statins reduce this IL-6-induced phosphorylation of STAT3 in hepatocytes. CONCLUSIONS: These results demonstrate that statins reduce IL-6-induced CRP production directly in hepatocytes via inhibition of protein geranylgeranylation. We further show that statins act via inhibition of STAT3 phosphorylation. These findings furnish new evidence for direct antiinflammatory properties of statins and provide new mechanistic insight into their clinical benefits. PMID- 15790935 TI - Myeloperoxidase and cardiovascular disease. AB - Myeloperoxidase (MPO) is a leukocyte-derived enzyme that catalyzes the formation of a number of reactive oxidant species. In addition to being an integral component of the innate immune response, evidence has emerged that MPO-derived oxidants contribute to tissue damage during inflammation. MPO-catalyzed reactions have been attributed to potentially proatherogenic biological activities throughout the evolution of cardiovascular disease, including during initiation, propagation, and acute complication phases of the atherosclerotic process. As a result, MPO and its downstream inflammatory pathways represent attractive targets for both prognostication and therapeutic intervention in the prophylaxis of atherosclerotic cardiovascular disease. PMID- 15790936 TI - Rho kinase inhibition and vascular protection: support from studies in Bartter and Gitelman syndrome. PMID- 15790937 TI - Association between IL-1beta gene polymorphism and myocardial infarction. PMID- 15790938 TI - Oxidative stress, antioxidants, and cardiovascular disease. PMID- 15790939 TI - Inorganic pyrophosphate: a paracrine regulator of vascular calcification and smooth muscle phenotype. PMID- 15790940 TI - When interleukin-18 conducts, the Preludio sounds the same no matter who plays. PMID- 15790941 TI - Clinical trial registration: a statement from the International Committee of Medical Journal Editors. PMID- 15790942 TI - High rate of early restenosis after carotid eversion endarterectomy in homozygous carriers of the normal mannose-binding lectin genotype. AB - BACKGROUND AND PURPOSE: Mannose-binding lectin (MBL) is thought to influence the pathophysiology of cardiovascular disease by decreasing the risk of advanced atherosclerosis and by contributing to enhanced ischemia reperfusion injury. Thus, we investigated the role of MBL in restenosis after eversion endarterectomy in patients with severe carotid atherosclerosis. METHODS: In a prospective study, 123 patients who underwent carotid endarterectomy were followed-up by carotid duplex scan (CDS) sonography for 14 months. In a retrospective study, we examined 17 patients and 29 patients, respectively, who had or had not at least 50% restenosis 29 months after carotid eversion endarterectomy. MBL genotypes were analyzed by a polymerase chain reaction-based method, and MBL serum concentrations were measured. RESULTS: In the prospective study in the patients homozygous for the normal MBL genotype, CDS values were significantly higher after 14 months of follow-up compared with the values measured 6 weeks after surgery (P<0.001). In contrast, only a slight increase was registered in patients carrying MBL variant alleles. The differences were much more pronounced in female than in male patients. Similar differences were observed when patients with high and low MBL serum concentrations were compared. In the retrospective study, a significant increase in the frequency of MBL variant genotypes was observed in patients not experiencing restenosis compared with the patients with restenosis (P=0.007). CONCLUSIONS: These results indicate that reoccurrence of stenosis after carotid endarterectomy is partially genetically determined and imply that MBL contributes significantly to the pathophysiology of this condition. PMID- 15790943 TI - Perfusion-weighted imaging/diffusion-weighted imaging mismatch on MRI can now be used to select patients for recombinant tissue plasminogen activator beyond 3 hours: con. PMID- 15790944 TI - Perfusion-weighted imaging/diffusion-weighted imaging mismatch on MRI can now be used to select patients for recombinant tissue plasminogen activator beyond 3 hours: pro. PMID- 15790946 TI - Magnesium sulfate in aneurysmal subarachnoid hemorrhage: a randomized controlled trial. AB - BACKGROUND AND PURPOSE: Magnesium reverses cerebral vasospasm and reduces infarct volume after experimental subarachnoid hemorrhage (SAH) in rats. We aimed to assess whether magnesium reduces the frequency of delayed cerebral ischemia (DCI) in patients with aneurysmal SAH. METHODS: Patients were randomized within 4 days after SAH. Magnesium sulfate therapy consisted of a continuous intravenous dose of 64 mmol/L per day, to be started within 4 days after SAH and continued until 14 days after occlusion of the aneurysm. The primary outcome DCI (defined as the occurrence of a new hypodense lesion on computed tomography compatible with clinical features of DCI) was analyzed according to the "on-treatment" principle. For the secondary outcome measures "poor outcome" (Rankin >3) and "excellent outcome" (Rankin 0), we used the "intention-to-treat" principle. RESULTS: A total of 283 patients were randomized. Magnesium treatment reduced the risk of DCI by 34% (hazard ratio, 0.66; 95% CI, 0.38 to 1.14). After 3 months, the risk reduction for poor outcome was 23% (risk ratio, 0.77; 95% CI, 0.54 to 1.09). At that time, 18 patients in the treatment group and 6 in the placebo group had an excellent outcome (risk ratio, 3.4; 95% CI, 1.3 to 8.9). CONCLUSIONS: This study suggests that magnesium reduces DCI and subsequent poor outcome, but the results are not yet definitive. A next step should be a phase III trial to confirm the beneficial effect of magnesium therapy, with poor outcome as primary outcome. PMID- 15790947 TI - Racial variations in location and risk of intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Risk factors for intracerebral hemorrhage (ICH) vary by location. Incidence rates of ICH are known to be higher in American blacks than whites, but how rates may differ by hemorrhage location is unknown. We sought to define incidence rates for different ICH locations in a biracial population. METHODS: All hospitalized patients age > or =20 years with spontaneous ICH were identified in the Greater Cincinnati/Northern Kentucky metropolitan area from May 1998 to July 2001 and August 2002 to April 2003. Incidence rates per 100,000 persons were age, sex, and race adjusted as appropriate to the 2000 US population. Risk ratios (RRs) with 95% CIs were calculated from unadjusted incidence rates. RESULTS: There were 1038 qualifying ICHs. Annual incidence rates per 100,000 persons > or =20 years of age were 48.9 for blacks and 26.6 for whites. Annual incidence rates per 100,000 blacks in lobar, deep cerebral, brain stem, and cerebellar locations were 15.2, 25.7, 5.1, and 2.9, respectively. Annual incidence rates per 100,000 whites in the same locations were 9.4, 13.0, 1.3, and 2.9. The greatest excess risk of ICH in blacks compared with whites was found among young to middle-aged (35 to 54 years) persons with brain stem (RR, 9.8; 95% CI, 4.2 to 23.0) and deep cerebral (RR, 4.5; 3.0 to 6.8) hemorrhage. CONCLUSIONS: The excess risk of ICH in American blacks is largely attributable to higher hemorrhage rates in young and middle-aged persons, particularly for deep cerebral and brain stem locations. Hypertension is the predominant risk factor for hemorrhages in these locations. PMID- 15790948 TI - Measuring accuracy of discharge diagnoses for a region-wide surveillance of hospitalized strokes. AB - BACKGROUND AND PURPOSE: Measuring the impact of stroke through population-based stroke registers is complex and costly. The aim of the present study is to assess the validity of hospital discharge diagnoses (all ages) and to estimate the total number of hospitalized stroke events in the Veneto region (Northeastern Italy, 4,500,000 inhabitants). METHODS: All discharges covering a 1-year period (1999) from Veneto hospitals with International Classification of Diseases, 9th Revision codes 342, 430 to 434, and 436 to 438 were identified. A stratified sample was extracted and submitted to retrospective clinical record review according to the World Health Organization MONItoring trends and determinants in CArdiovascular disease stroke project. Using the positive predictive value (PPV) for validated acute stroke of each code to adjust for inaccuracy of discharge diagnoses, an estimate of hospital strokes was obtained. RESULTS: 4015 admissions were reviewed. Codes 430, 431, 434, and 436 as primary diagnoses had the highest PPV, which sharply decreased in the other diagnostic levels. Code 342 also showed a high PPV. The probability of suspected events meeting the stroke definition increased with age and was highest for patients admitted to neurological wards and for fatal events. Overall 9400 strokes (first-ever and recurrent) were estimated to be hospitalized in 1999, with an attack rate of 208 per 100,000. CONCLUSIONS: Our data indicate that once validation studies are undertaken on a sample of all hospitalized events, hospital discharge records can provide a valuable source of information on the actual burden of strokes on hospital services. PMID- 15790949 TI - Neuroprotective effect of darbepoetin alfa, a novel recombinant erythropoietic protein, in focal cerebral ischemia in rats. AB - BACKGROUND AND PURPOSE: Darbepoetin alfa is a novel erythropoiesis-stimulating protein developed for treating anemia. In animal models, exogenous recombinant human erythropoietin has been reported to be beneficial in treating experimental cerebral ischemia. In this study, we determined whether darbepoetin alfa would protect in a rat model of transient focal cerebral ischemia. METHODS: Rats received 2-hour middle cerebral artery suture-occlusion. The drug (darbepoetin alfa, 10 microg/kg) or vehicle was administered intraperitoneally 2 hours after onset of middle cerebral artery occlusion. Animals were allowed to survive for 3 or 14 days. Behavioral tests were performed sequentially. Infarct volumes and brain swelling were determined. RESULTS: Darbepoetin alfa-treated rats showed improved neuroscores relative to vehicle-treated animals beginning within 1 hour of treatment and persisting throughout the 14-day survival period. Darbepoetin alfa significantly reduced corrected total (cortical + subcortical) infarct volume (56.3+/-20.6 and 110.8+/-6.8 mm3, respectively) and total infarct areas at multiple levels compared with vehicle in the 14-day survival group. Brain swelling was not affected by treatment. CONCLUSIONS: Darbepoetin alfa confers behavioral and histological neuroprotection after focal ischemia in rats. PMID- 15790950 TI - Does the mismatch match the penumbra? Magnetic resonance imaging and positron emission tomography in early ischemic stroke. AB - BACKGROUND AND PURPOSE: In ischemic stroke, diffusion-weighted (DW) and perfusion weighted (PW) magnet resonance imaging (MRI) is used to define the mismatch as the therapeutic target. With positron emission tomography (PET), we characterized the metabolic patterns of tissue compartments identified by MRI and compared the volumes of mismatch to those of PET-defined penumbra. METHODS: In 6 acute (median, 5.2 hours) and 7 chronic (median, 10 days) stroke patients in whom a mismatch was defined by PW/DW MRI, PET was performed (median, 120-minute delay). Cerebral blood flow (CBF), oxygen metabolism (CMRO2), and oxygen extraction fraction (OEF) was determined in the areas of DWI lesion, mismatch, and oligemia. Then, the mismatch volume was compared with the volume of penumbra. RESULTS: DWI lesions showed impaired tissue integrity (low CMRO2 and low OEF). Mismatch areas were viable (normal CMRO2) but showed largely varying OEF. Oligemic areas had metabolic patterns comparable to normal tissue. A mismatch volume was found in all 13 patients. However, only 8 of 13 had a corresponding penumbra volume that covered only a part of the mismatch. CONCLUSIONS: Our comparative PET/MRI study confirmed the current pathophysiological hypothesis for the DWI lesion and for the oligemic areas. However, the mismatch area did not reliably detect elevated OEF and overestimated the penumbra defined by PET. PMID- 15790951 TI - Essential role of a Ca2+-selective, store-operated current (ISOC) in endothelial cell permeability: determinants of the vascular leak site. AB - Store-operated calcium (SOC) entry is sufficient to disrupt the extra-alveolar, but not the alveolar, endothelial cell barrier. Mechanism(s) underlying such insensitivity to transitions in cytosolic calcium ([Ca2+]i) in microvascular endothelial cells are unknown. Depletion of stored Ca2+ activates a larger SOC entry response in extra-alveolar (pulmonary artery; PAECs) than alveolar (pulmonary microvascular; PMVECs) endothelial cells. In vivo permeation studies revealed that Ca2+ store depletion activates similar nonselective cationic conductances in PAECs and PMVECs, while only PAECs possess the calcium-selective, store-operated Ca2+ entry current, I(SOC). Pretreatment with the type 4 phosphodiesterase inhibitor, rolipram, abolished thapsigargin-activated I(SOC) in PAECs, and revealed I(SOC) in PMVECs. Rolipram pretreatment shifted the thapsigargin-induced fluid leak site from extra-alveolar to alveolar vessels in the intact pulmonary circulation. Thus, our results indicate I(SOC) provides a [Ca2+]i source that is needed to disrupt the endothelial cell barrier, and demonstrate that intracellular events controlling I(SOC) activation coordinate the site-specific vascular response to inflammation. PMID- 15790952 TI - Adenoviral gene transfer of mutant phospholamban rescues contractile dysfunction in failing rabbit myocytes with relatively preserved SERCA function. AB - In heart failure (HF) a main factor in reduced contractility is reduced SR Ca2+ content and reversed force-frequency response (FFR), ie, from positive to negative. Our arrhythmogenic rabbit HF model exhibits decreased contractility mainly due to an increase in Na/Ca exchange (NCX) activity (with only modest decrease in SR Ca2+-ATPase (SERCA) function), similar to many end-stage HF patients. Here we test whether phospholamban (PLB) inhibition using a dominant negative mutant PLB adenovirus (K3E/R14E, AdPLB-dn, with beta-galactosidase adenovirus as control) could enhance SERCA function and restore Ca2+ transients and positive FFR in ventricular myocytes from these HF rabbits. HF myocytes infected with AdPLB-dn (versus control) had enhanced Ca2+ transient amplitude (2.0+/-0.1 versus 1.6+/-0.05 F/Fo at 0.5 Hz, P<0.05) and had a positive FFR, whereas acutely isolated HF myocytes or those infected with Adbetagal had negative FFR. Ca2+ transients declined faster in AdPLB-dn versus Adbetagal myocytes (RT50%: 317+/-29 versus 551+/-90 ms at 0.5 Hz, P<0.05) and had an increased SR Ca2+ load (3.5+/-0.3 versus 2.6+/-0.2 F/Fo at 0.5 Hz, P<0.05), indicative of increased SERCA function. Furthermore, this restoration of function was not due to changes in NCX or SERCA expression. Thus, increasing SERCA activity in failing myocytes by AdPLB-dn gene transfer reversed the contractile dysfunction (and restored positive FFR) by increasing SR Ca2+ load. This approach could enhance contractile function in failing hearts of various etiologies, even here where reduced SERCA activity is not the main dysfunction. PMID- 15790953 TI - Targeted disruption of TGF-beta-Smad3 signaling leads to enhanced neointimal hyperplasia with diminished matrix deposition in response to vascular injury. AB - The role of transforming growth factor (TGF)-beta and its signal in atherogenesis is not fully understood. Here, we examined mice lacking Smad3, a major downstream mediator of TGF-beta, to clarify the precise role of Smad3-dependent signaling in vascular response to injury. Femoral arteries were injured in wild-type and Smad3 null (null) male mice on C57Bl/6 background. Histopathological evaluation of the arteries 1 to 3 weeks after the injury revealed significant enhancement of neointimal hyperplasia in null compared with wild-type mice. Transplantation of null bone marrow to wild-type mice did not enhance neointimal thickening, suggesting that vascular cells in situ play a major role in the response. Null intima contained more proliferating smooth muscle cells (SMC) with less amount of collagen compared with wild-type intima. TGF-beta caused significant inhibition of cellular proliferation in wild-type aortic SMC, whereas the growth of null SMC was only weakly inhibited by TGF-beta in vitro, indicating a crucial role of Smad3 in the growth inhibitory function. On the other hand, Smad3-deficiency did not attenuate chemotaxis of SMC toward TGF-beta. TGF-beta increased transcript level of alpha2 type I collagen and tissue inhibitor of metalloproteinases-1, and suppressed expression and activity of matrix metalloproteinases in wild-type SMC. However, these effects of TGF-beta were diminished in null SMC. Our findings altogether show that the loss of Smad3 pathway causes enhanced neointimal hyperplasia on injury through modulation of growth and matrix regulation in vascular SMC. These results indicate a vasculoprotective role of endogenous Smad3 in response to injury. PMID- 15790954 TI - AMP-activated protein kinase signaling stimulates VEGF expression and angiogenesis in skeletal muscle. AB - AMP-activated protein kinase (AMPK) is regulated by various cellular stresses. Vascular endothelial growth factor (VEGF), a key regulator of angiogenesis, is also upregulated by several stress-inducible factors such as hypoxia and stimulation by cytokines and growth factors. Here, we investigated whether AMPK signaling in muscle has a role in regulating VEGF-mediated angiogenic processes. AICAR stimulated VEGF mRNA and protein levels in C2C12 myotube cultures. Transduction with dominant-negative AMPK abolished AICAR-induced VEGF expression at both steady state mRNA and protein levels. AICAR increased VEGF mRNA stability without affecting VEGF promoter activity. AICAR also stimulated p38 mitogen activated protein kinase (p38 MAPK) phosphorylation. Activation of p38 MAPK was suppressed by transduction with dominant-negative AMPK, suggesting that AMPK is upstream of p38 MAPK. The p38 MAPK inhibitor SB203580 blocked AICAR-induced increase in VEGF mRNA and protein levels, indicating that AICAR-mediated VEGF induction is dependent on p38 MAPK signaling. AICAR treatment increased VEGF expression and accelerated angiogenic repair of ischemic hindlimbs in mice in an AMPK-dependent manner. These data indicate that AMPK-p38 MAPK signaling cascade can increase VEGF production in muscle and promote angiogenesis in response to ischemic injury. PMID- 15790955 TI - Liver x receptor agonists inhibit cytokine-induced osteopontin expression in macrophages through interference with activator protein-1 signaling pathways. AB - Osteopontin (OPN) is a proinflammatory cytokine and adhesion molecule implicated in the chemoattraction of monocytes and in cell-mediated immunity. We have recently reported that genetic OPN-deficiency attenuates the development of atherosclerosis in apoE-/- mice identifying OPN as potential target for pharmacological intervention in atherosclerosis. Synthetic agonists for the Liver X Receptor (LXR), members of the nuclear hormone receptor superfamily, prevent the development of atherosclerosis by regulating cholesterol homeostasis and suppressing inflammatory gene expression in macrophages. We demonstrate here that LXR ligands inhibit cytokine-induced OPN expression in macrophages. Two synthetic LXR ligands, T0901317 and GW3965, inhibited TNF-alpha, IL-1beta, INF-gamma and lipopolysaccharide induced OPN mRNA and protein expression in RAW 264.7 macrophages. Transient transfection experiments revealed that LXR ligands suppress cytokine-induced OPN promoter activity. Deletion analysis, heterologous promoter assays, and site-directed mutagenesis identified an activator protein-1 (AP-1) consensus site at -76 relative to the initiation site that supports OPN transcription in macrophages and mediates the effects of LXR ligands to inhibit OPN transcription. Electrophoretic mobility shift and chromatin immunoprecipitation assays indicated that LXR agonists inhibit cytokine-induced c Fos and phospho-c-Jun binding to this AP-1 site. Cytokine-induced c-Fos and phospho-c-Jun protein expression was inhibited by LXR ligands and overexpression of c-Fos and c-Jun reversed the inhibitory effect of LXR ligands on OPN promoter activity in transactivation assays. Finally, treatment of C57BL/6J mice with LXR ligands inhibited OPN expression in peritoneal macrophages indicating that the observed effects of LXR ligands to inhibit OPN expression are applicable in vivo. These observations identify the regulation of macrophage OPN expression as a mechanism whereby LXR ligands may impact macrophage inflammatory responses and atherosclerosis. The full text of this article is available online at http://circres.ahajournals.org. PMID- 15790956 TI - [Ca2+]i reduction increases cellular proliferation and apoptosis in vascular smooth muscle cells: relevance to the ADPKD phenotype. AB - Cardiovascular complications are the leading cause of morbidity and mortality in autosomal dominant polycystic kidney disease. Pkd2+/- vascular smooth muscle cells (VSMCs) have an abnormal phenotype and defective intracellular Ca2+ ([Ca2+]i) regulation. We examined cAMP content in vascular smooth muscles from Pkd2+/- mice because cAMP is elevated in cystic renal epithelial cells. We found cAMP concentration was significantly increased in Pkd2+/- vessels compared with wild-type vessels. Furthermore, reducing the wild-type VSMC [Ca2+]i by Verapamil or BAPTA-AM significantly increased cellular cAMP concentration (mainly by phosphodiesterase [PDE] inhibition), the rate of VSMC proliferation (determined by direct cell counting, 3H-incorporation, FACS analysis of cells entering S phase, and quantitative Western PCNA and ERK1/2 analyses), and the rate of apoptosis (by Hoechst staining, FACS analysis of the Annexin-V positive cells, and quantitative Western Bax, cytochrome c, and activated caspase 9 and 3 analyses). The low [Ca2+]i induced VSMC proliferation was independent of cAMP/B Raf signaling, while that of apoptosis was promoted by cAMP. In summary, Pkd2+/- VSMCs have elevated cAMP levels. This elevation can also be induced by reducing [Ca2+]i in wild-type VSMCs. The [Ca2+]i reduction and cAMP accumulation can cause an increase in both cellular proliferation and apoptosis, resembling Pkd mutant phenotype. PMID- 15790957 TI - Characterization of a novel PKA phosphorylation site, serine-2030, reveals no PKA hyperphosphorylation of the cardiac ryanodine receptor in canine heart failure. AB - Hyperphosphorylation of the cardiac Ca2+ release channel (ryanodine receptor, RyR2) by protein kinase A (PKA) at serine-2808 has been proposed to be a key mechanism responsible for cardiac dysfunction in heart failure (HF). However, the sites of PKA phosphorylation in RyR2 and their phosphorylation status in HF are not well defined. Here we used various approaches to investigate the phosphorylation of RyR2 by PKA. Mutating serine-2808, which was thought to be the only PKA phosphorylation site in RyR2, did not abolish the phosphorylation of RyR2 by PKA. Two-dimensional phosphopeptide mapping revealed two major PKA phosphopeptides, one of which corresponded to the known serine-2808 site. Another, novel, PKA phosphorylation site, serine 2030, was identified by Edman sequencing. Using phospho-specific antibodies, we showed that the novel serine 2030 site was phosphorylated in rat cardiac myocytes stimulated with isoproterenol, but not in unstimulated cells, whereas serine-2808 was considerably phosphorylated before and after isoproterenol treatment. We further showed that serine-2030 was stoichiometrically phosphorylated by PKA, but not by CaMKII, and that mutations of serine-2030 altered neither the FKBP12.6-RyR2 interaction nor the Ca2+ dependence of [3H]ryanodine binding. Moreover, the levels of phosphorylation of RyR2 at serine-2030 and serine-2808 in both failing and non-failing canine hearts were similar. Together, our data indicate that serine-2030 is a major PKA phosphorylation site in RyR2 responding to acute beta adrenergic stimulation, and that RyR2 is not hyperphosphorylated by PKA in canine HF. PMID- 15790958 TI - Homeobox protein Hop functions in the adult cardiac conduction system. AB - Hop is an unusual homeobox gene expressed in the embryonic and adult heart. Hop acts downstream of Nkx2-5 during development, and Nkx2-5 mutations are associated with cardiac conduction system (CCS) defects. Inactivation of Hop in the mouse is lethal in half of the expected null embryos. Here, we show that Hop is expressed strongly in the adult CCS. Hop-/- adult mice display conduction defects below the atrioventricular node (AVN) as determined by invasive electrophysiological testing. These defects are associated with decreased expression of connexin40. Our results suggest that Hop functions in the adult CCS and demonstrate conservation of molecular hierarchies between embryonic myocardium and the specialized conduction tissue of the mature heart. PMID- 15790959 TI - Unmasking hypertension. PMID- 15790960 TI - Pulse pressure and effects of losartan or atenolol in patients with hypertension and left ventricular hypertrophy. AB - In the Losartan Intervention For Endpoint reduction in hypertension (LIFE) study, the primary composite end point of cardiovascular death, stroke, and myocardial infarction was reduced by losartan versus atenolol in patients with hypertension and left ventricular hypertrophy. The objective of this post hoc analysis was to determine the influence of pulse pressure on outcome. Patients were divided into quartiles of baseline pulse pressure. Cox regression, including baseline Framingham risk score as a covariate, was used to compare risk in the quartiles. In the atenolol group, there were significantly higher risks in the highest versus lowest quartile for the composite end point 28% (confidence interval [CI], 2% to 62%; P=0.035), stroke 84% (CI, 32% to 157%; P<0.001), and total mortality 41% (CI, 7% to 84%; P=0.013). Risk for myocardial infarction was 44% higher (CI, 5% to 120%; P=0.089). The risks in the losartan group also increased with increasing quartile, but were lower than in the atenolol group, and differences between the highest and lowest quartiles were not significant: composite end point 12% (CI, -13% to 44%; P>0.2), stroke -5% (CI, -34% to 37%; P>0.2), myocardial infarction 30% (CI, -13% to 94%; P>0.2), and total mortality 32% (CI, 1% to 76%; P=0.062). In patients with hypertension and left ventricular hypertrophy in the LIFE study, there were significantly higher risks, adjusted for the Framingham risk score, for the primary composite end point, stroke, and total mortality in the highest versus lowest quartile of pulse pressure with atenolol-based treatment. The risks in the losartan group also increased with increasing pulse pressure quartile, but were lower than those in the atenolol group, and were not significant. PMID- 15790961 TI - Clinical trial registration: a statement from the International Committee of Medical Journal Editors. PMID- 15790962 TI - The roles of receptor and ligand endocytosis in regulating Notch signaling. AB - Cell-cell signaling is a central process in the formation of multicellular organisms. Notch (N) is the receptor of a conserved signaling pathway that regulates numerous developmental decisions, and the misregulation of N has been linked to various physiological and developmental disorders. The endocytosis of N and its ligands is a key mechanism by which N-mediated cell-cell signaling is developmentally regulated. We review here the recent findings that have highlighted the importance and complexity of this regulation. PMID- 15790963 TI - Stabilization of the retinal vascular network by reciprocal feedback between blood vessels and astrocytes. AB - Development of the retinal vasculature is controlled by a hierarchy of interactions among retinal neurons, astrocytes and blood vessels. Retinal neurons release platelet-derived growth factor (PDGFA) to stimulate proliferation of astrocytes, which in turn stimulate blood vessel growth by secreting vascular endothelial cell growth factor (VEGF). Presumably, there must be counteractive mechanisms for limiting astrocyte proliferation and VEGF production to prevent runaway angiogenesis. Here, we present evidence that the developing vessels provide feedback signals that trigger astrocyte differentiation--marked by cessation of cell division, upregulation of glial fibrillary acidic protein (GFAP) and downregulation of VEGF. We prevented retinal vessel development by raising newborn mice in a high-oxygen atmosphere, which leads, paradoxically, to retinal hypoxia (confirmed by using the oxygen-sensing reagent EF5). The forced absence of vessels caused prolonged astrocyte proliferation and inhibited astrocyte differentiation in vivo. We could reproduce these effects by culturing retinal astrocytes in a low oxygen atmosphere, raising the possibility that blood borne oxygen itself might induce astrocyte differentiation and indirectly prevent further elaboration of the vascular network. PMID- 15790964 TI - Components of the transcriptional Mediator complex are required for asymmetric cell division in C. elegans. AB - Asymmetric cell division is a fundamental process that produces cellular diversity during development. In C. elegans, the Wnt signaling pathway regulates the asymmetric divisions of a number of cells including the T blast cell. We found that the let-19 and dpy-22 mutants have defects in their T-cell lineage, and lineage analyses showed that the defects were caused by disruption in the asymmetry of the T-cell division. We found that let-19 and dpy-22 encode homologs of the human proteins MED13/TRAP240 and MED12/TRAP230, respectively, which are components of the Mediator complex. Mediator is a multi-component complex that can regulate transcription by transducing the signals between activators and RNA polymerase in vitro. We also showed that LET-19 and DPY-22 form a complex in vivo with other components of Mediator, SUR-2/MED23 and LET-425/MED6. In the let-19 and dpy-22 mutants, tlp-1, which is normally expressed asymmetrically between the T-cell daughters through the function of the Wnt pathway, was expressed symmetrically in both daughter cells. Furthermore, we found that the let-19 and dpy-22 mutants were defective in the fusion of the Pn.p cell, a process that is regulated by bar-1/beta-catenin. Ectopic cell fusion in bar-1 mutants was suppressed by the let-19 or dpy-22 mutations, while defective cell fusion in let 19 mutants was suppressed by lin-39/Hox mutations, suggesting that let-19 and dpy 22 repress the transcription of lin-39. These results suggest that LET-19 and DPY 22 in the Mediator complex repress the transcription of Wnt target genes. PMID- 15790965 TI - Functional analysis of the chicken delta1-crystallin enhancer activity in Drosophila reveals remarkable evolutionary conservation between chicken and fly. AB - Functional conservation of enhancers among evolutionarily diverged organisms is a powerful way to identify basic regulatory circuits and key developmental regulators. This is especially applicable to Crystallin genes. Despite unexpected heterogeneity and diversity in their DNA sequences, many studies have revealed that most of the Crystallin genes are regulated by a relatively small set of developmentally important transcription factors. The chicken delta1-crystallin is one of the best-characterized Crystallin genes. Its lens-specific regulation is governed by a 30 bp long DC5 fragment present in the third intron of the gene. DC5 contains PAX6 and SOX2 binding sites, and its activity depends on the cooperative binding of these two transcription factors. To test the idea that Pax6 and Sox2, together with the DC5 enhancer, could form a basic regulatory circuit functional in distantly related animals, we introduced the DC5 fragment into Drosophila and studied its activation pattern and regulation. The results show that the DC5 enhancer is not only active in the compound eye but, remarkably, is specifically active in those cells responsible for Crystallin secretion in Drosophila, i.e. the cone cells. However, regulation of the DC5 enhancer is carried out not by Pax6, but by Pax2 (D-Pax2; shaven--FlyBase) in combination with the Sox2 homologue SoxN. Both proteins (D-PAX2 and SOXN) bind cooperatively to the DC5 fragment and activate the enhancer synergistically. As PAX6 and PAX2 proteins derive from the same ancestor, we propose that during evolution Pax6 function in vertebrate lens development was retained by Pax2 in Drosophila. PMID- 15790966 TI - Cilia-driven fluid flow in the zebrafish pronephros, brain and Kupffer's vesicle is required for normal organogenesis. AB - Cilia, as motile and sensory organelles, have been implicated in normal development, as well as diseases including cystic kidney disease, hydrocephalus and situs inversus. In kidney epithelia, cilia are proposed to be non-motile sensory organelles, while in the mouse node, two cilia populations, motile and non-motile have been proposed to regulate situs. We show that cilia in the zebrafish larval kidney, the spinal cord and Kupffer's vesicle are motile, suggesting that fluid flow is a common feature of each of these organs. Disruption of cilia structure or motility resulted in pronephric cyst formation, hydrocephalus and left-right asymmetry defects. The data show that loss of fluid flow leads to fluid accumulation, which can account for organ distension pathologies in the kidney and brain. In Kupffer's vesicle, loss of flow is associated with loss of left-right patterning, indicating that the 'nodal flow' mechanism of generating situs is conserved in non-mammalian vertebrates. PMID- 15790967 TI - Analysis of xbx genes in C. elegans. AB - Cilia and flagella are widespread eukaryotic subcellular components that are conserved from green algae to mammals. In different organisms they function in cell motility, movement of extracellular fluids and sensory reception. While the function and structural description of cilia and flagella are well established, there are many questions that remain unanswered. In particular, very little is known about the developmental mechanisms by which cilia are generated and shaped and how their components are assembled into functional machineries. To find genes involved in cilia development we used as a search tool a promoter motif, the X box, which participates in the regulation of certain ciliary genes in the nematode Caenorhabditis elegans. By using a genome search approach for X-box promoter motif-containing genes (xbx genes) we identified a list of about 750 xbx genes (candidates). This list comprises some already known ciliary genes as well as new genes, many of which we hypothesize to be important for cilium structure and function. We derived a C. elegans X-box consensus sequence by in vivo expression analysis. We found that xbx gene expression patterns were dependent on particular X-box nucleotide compositions and the distance from the respective gene start. We propose a model where DAF-19, the RFX-type transcription factor binding to the X-box, is responsible for the development of a ciliary module in C. elegans, which includes genes for cilium structure, transport machinery, receptors and other factors. PMID- 15790968 TI - Regulation of chemosensory and GABAergic motor neuron development by the C. elegans Aristaless/Arx homolog alr-1. AB - Mutations in the highly conserved Aristaless-related homeodomain protein ARX have been shown to underlie multiple forms of X-linked mental retardation. Arx knockout mice exhibit thinner cerebral cortices because of decreased neural precursor proliferation, and also exhibit defects in the differentiation and migration of GABAergic interneurons. However, the role of ARX in the observed behavioral and developmental abnormalities is unclear. The regulatory functions of individual homeodomain proteins and the networks in which they act are frequently highly conserved across species, although these networks may be deployed in different developmental contexts. In Drosophila, aristaless mutants exhibit defects in the development of terminal appendages, and Aristaless has been shown to function with the LIM-homeodomain protein LIM1 to regulate leg development. Here, we describe the role of the Aristaless/Arx homolog alr-1 in C. elegans. We show that alr-1 acts in a pathway with the LIM1 ortholog lin-11 to regulate the development of a subset of chemosensory neurons. Moreover, we demonstrate that the differentiation of a GABAergic motoneuron subtype is affected in alr-1 mutants, suggesting parallels with ARX functions in vertebrates. Investigating ALR-1 functions in C. elegans may yield insights into the role of this important protein in neuronal development and the etiology of mental retardation. PMID- 15790969 TI - A subset of oligodendrocytes generated from radial glia in the dorsal spinal cord. AB - Many oligodendrocytes in the spinal cord are derived from a region of the ventral ventricular zone (VZ) that also gives rise to motoneurons. Cell fate specification in this region depends on sonic hedgehog (Shh) from the notochord and floor plate. There have been suggestions of an additional source(s) of oligodendrocytes in the dorsal spinal cord. We revisited this idea by Cre-lox fate-mapping in transgenic mice. We found that a subpopulation of oligodendrocytes is generated from the Dbx1-expressing domain of the VZ, spanning the dorsoventral midline. Dbx-derived oligodendrocytes comprise less than 5% of the total; they are formed late during embryogenesis by transformation of radial glia and settle mainly in the lateral white matter. Development of Dbx-derived oligodendrocytes in vitro can occur independently of Shh but requires FGF signalling. Dbx-expressing precursors also generate astrocytes and interneurons, but do not contribute to the ependymal layer of the postnatal spinal cord. PMID- 15790970 TI - Tbx3 can alter limb position along the rostrocaudal axis of the developing embryo. AB - The limbs of the vertebrate embryo form at precise locations along the body and these positions are fixed across different species. The mechanisms that control this process are not understood. Ectopic expression of Tbx3, a transcriptional repressor that belongs to the Tbx2/3/4/5 subfamily of T-box transcriptional regulators, in the forelimb results in a rostral shift in the position of the limb along the main body axis. By contrast, a transcriptional activator form of Tbx3 shifts the limb to more caudal locations. We also show that dHand and Gli3, genes previously implicated in anteroposterior pre-patterning of the limb-forming region, are also involved in refining the position of the limbs. Our data suggest a new role for Tbx3 in positioning the limb along the main body axis through a genetic interplay between dHand and Gli3. PMID- 15790971 TI - New regulatory interactions and cellular responses in the isthmic organizer region revealed by altering Gbx2 expression. AB - The mouse homeobox gene Gbx2 is first expressed throughout the posterior region of the embryo during gastrulation, and becomes restricted to rhombomeres 1-3 (r1 3) by embryonic day 8.5 (E8.5). Previous studies have shown that r1-3 do not develop in Gbx2 mutants and that there is an early caudal expansion of the midbrain gene Otx2 to the anterior border of r4. Furthermore, expression of Wnt1 and Fgf8, two crucial components of the isthmic organizer, is no longer segregated to adjacent domains in Gbx2 mutants. In this study, we extend the phenotypic analysis of Gbx2 mutants by showing that Gbx2 is not only required for development of r1-3, but also for normal gene expression in r4-6. To determine whether Gbx2 can alter hindbrain development, we generated Hoxb1-Gbx2 (HG) transgenic mice in which Gbx2 is ectopically expressed in r4. We show that Gbx2 is not sufficient to induce r1-3 development in r4. To test whether an Otx2/Gbx2 interface can induce r1-3 development, we introduced the HG transgene onto a Gbx2 null mutant background and recreated a new Otx2/Gbx2 border in the anterior hindbrain. Development of r3, but not r1 and r2, is rescued in Gbx2-/-; HG embryos. In addition, the normal spatial relationship of Wnt1 and Fgf8 is established at the new Otx2/Gbx2 border, demonstrating that an interaction between Otx2 and Gbx2 is sufficient to produce the normal pattern of Wnt1 and Fgf8 expression. However, the expression domains of Fgf8 and Spry1, a downstream target of Fgf8, are greatly reduced in mid/hindbrain junction area of Gbx2-/-; HG embryos and the posterior midbrain is truncated because of abnormal cell death. Interestingly, we show that increased cell death and a partial loss of the midbrain are associated with increased expression of Fgf8 and Spry1 in Gbx2 conditional mutants that lack Gbx2 in r1 after E9.0. These results together suggest that cell survival in the posterior midbrain is positively or negatively regulated by Fgf8, depending on Fgf8 expression level. Our studies provide new insights into the regulatory interactions that maintain isthmic organizer gene expression and the consequences of altered levels of organizer gene expression on cell survival. PMID- 15790972 TI - The Abelson tyrosine kinase, the Trio GEF and Enabled interact with the Netrin receptor Frazzled in Drosophila. AB - The attractive Netrin receptor Frazzled (Fra), and the signaling molecules Abelson tyrosine kinase (Abl), the guanine nucleotide-exchange factor Trio, and the Abl substrate Enabled (Ena), all regulate axon pathfinding at the Drosophila embryonic CNS midline. We detect genetic and/or physical interactions between Fra and these effector molecules that suggest that they act in concert to guide axons across the midline. Mutations in Abl and trio dominantly enhance fra and Netrin mutant CNS phenotypes, and fra;Abl and fra;trio double mutants display a dramatic loss of axons in a majority of commissures. Conversely, heterozygosity for ena reduces the severity of the CNS phenotype in fra, Netrin and trio,Abl mutants. Consistent with an in vivo role for these molecules as effectors of Fra signaling, heterozygosity for Abl, trio or ena reduces the number of axons that inappropriately cross the midline in embryos expressing the chimeric Robo-Fra receptor. Fra interacts physically with Abl and Trio in GST-pulldown assays and in co-immunoprecipitation experiments. In addition, tyrosine phosphorylation of Trio and Fra is elevated in S2 cells when Abl levels are increased. Together, these data suggest that Abl, Trio, Ena and Fra are integrated into a complex signaling network that regulates axon guidance at the CNS midline. PMID- 15790973 TI - The role of Axin2 in calvarial morphogenesis and craniosynostosis. AB - Axin1 and its homolog Axin2/conductin/Axil are negative regulators of the canonical Wnt pathway that suppress signal transduction by promoting degradation of beta-catenin. Mice with deletion of Axin1 exhibit defects in axis determination and brain patterning during early embryonic development. We show that Axin2 is expressed in the osteogenic fronts and periosteum of developing sutures during skull morphogenesis. Targeted disruption of Axin2 in mice induces malformations of skull structures, a phenotype resembling craniosynostosis in humans. In the mutants, premature fusion of cranial sutures occurs at early postnatal stages. To elucidate the mechanism of craniosynostosis, we studied intramembranous ossification in Axin2-null mice. The calvarial osteoblast development is significantly affected by the Axin2 mutation. The Axin2 mutant displays enhanced expansion of osteoprogenitors, accelerated ossification, stimulated expression of osteogenic markers and increases in mineralization. Inactivation of Axin2 promotes osteoblast proliferation and differentiation in vivo and in vitro. Furthermore, as the mammalian skull is formed from cranial skeletogenic mesenchyme, which is derived from mesoderm and neural crest, our data argue for a region-specific effect of Axin2 on neural crest dependent skeletogenesis. The craniofacial anomalies caused by the Axin2 mutation are mediated through activation of beta-catenin signaling, suggesting a novel role for the Wnt pathway in skull morphogenesis. PMID- 15790975 TI - Is big beautiful? The continuing story of ADAM33 and asthma. PMID- 15790974 TI - Defective surfactant secretion in a mouse model of Hermansky-Pudlak syndrome. AB - Hermansky-Pudlak syndrome (HPS) in humans represents a family of disorders of lysosome-related organelle biogenesis associated with severe, progressive pulmonary disease. Human case reports and a mouse model of HPS, the pale ear/pearl mouse (ep/pe), exhibit giant lamellar bodies (GLB) in type II alveolar epithelial cells. We examined surfactant proteins and phospholipid from ep/pe mice to elucidate the process of GLB formation. The 2.8-fold enrichment of tissue phospholipids in ep/pe mice resulted from accumulation from birth through adulthood. Tissue surfactant protein (SP)-B and -C were increased in adult ep/pe mice compared with wild-type mice (WT), whereas SP-A and -D were not different. Large aggregate surfactant (LA) from adult ep/pe mice had decreased phospholipid, SP-B, and SP-C, with no differences in SP-A and -D compared with WT. Although LA from ep/pe animals exhibited an increased total protein-to-total phospholipid ratio compared with WT, surface tension was not compromised. Phospholipid secretion from isolated type II cells showed that basal and stimulated secretion from ep/pe cells were approximately 50% of WT cells. Together, our data indicate that GLB formation is not associated with abnormal trafficking or recycling of surfactant material. Instead, impaired secretion is an important component of GLB formation in ep/pe mice. PMID- 15790976 TI - Nitric oxide, hypoxia, and superoxide: the good, the bad, and the ugly! PMID- 15790977 TI - Symptoms and the early diagnosis of lung cancer. PMID- 15790978 TI - Lung function estimates in idiopathic pulmonary fibrosis: the potential for a simple classification. PMID- 15790980 TI - Contribution of ADAM33 polymorphisms to the population risk of asthma. AB - BACKGROUND: ADAM 33 is the first gene identified as a candidate for asthma by positional cloning techniques, with association studies reaching impressive statistical significance. It has a postulated role in myogenesis, airway modelling, and signalling via protein shedding. Concerns over the methodology of the initial study have led to several attempts at replication, with inconsistent results. METHOD: To clarify the role of ADAM33 in determining the risk of asthma in the general population, new transmission disequilibrium and case-control studies were undertaken followed by a meta-analysis of all existing data. RESULTS: Studies in Icelandic and UK populations revealed no association when taken in isolation. The meta-analysis, however, showed that the F+1 and ST+7 variants were significantly associated with asthma in both types of study. CONCLUSIONS: The additional risk imparted by this variation would account for 50,000 excess asthma cases in the UK alone. This study also demonstrates the size of study required to investigate such hypotheses adequately. PMID- 15790981 TI - Relationship of airway wall thickening to an imbalance between matrix metalloproteinase-9 and its inhibitor in asthma. AB - BACKGROUND: The balance between matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) may be critical in extracellular matrix remodelling, a characteristic of asthmatic airways. An excess of TIMP-1 over MMP 9 has been associated with chronic airflow obstruction but the mechanisms underlying this association remain unknown. Recent computed tomographic (CT) studies indicate that airway wall thickening is associated with chronic airflow obstruction. METHODS: Sputum levels of MMP-9, TIMP-1, and their molar ratio were examined in 26 patients with stable asthma and their relationship with pulmonary function and airway wall thickness, assessed by a validated CT technique which measured wall area corrected by body surface area (WA/BSA), the ratio of WA to outer wall area (WA%), and the absolute wall thickness corrected by radicalBSA of a segmental bronchus (T/ radicalBSA), was examined. RESULTS: Sputum MMP-9 levels were inversely correlated with WA% and TIMP-1 levels were positively correlated with WA/BSA and T/ radicalBSA. The MMP-9/TIMP-1 molar ratio was inversely correlated with WA% and T/ radicalBSA and positively correlated with post bronchodilator values of mid-forced expiratory flow and maximum expiratory flow at the quartile of lung volume. CONCLUSION: Excess TIMP-1 may have a pathogenetic role in airway wall thickening in asthmatic patients which may result in chronic airflow obstruction. PMID- 15790982 TI - Efficacy of low and high dose inhaled corticosteroid in smokers versus non smokers with mild asthma. AB - BACKGROUND: Cigarette smokers with asthma are insensitive to short term inhaled corticosteroid therapy, but efficacy when given for a longer duration at different doses is unknown. METHODS: Ninety five individuals with mild asthma were recruited to a multicentre, randomised, double blind, parallel group study comparing inhaled beclomethasone in doses of 400 microg or 2000 microg daily for 12 weeks in smokers and non-smokers. The primary end point was the change in morning peak expiratory flow (PEF). Secondary end points included evening PEF, use of reliever inhaler, number of asthma exacerbations, spirometric parameters, and asthma control score. RESULTS: After 12 weeks of inhaled beclomethasone there was a considerable difference between the morning PEF measurements of smokers and non-smokers with asthma (-18 (95% CI -35 to -1), adjusted p = 0.035). Among those receiving 400 microg daily there was a difference between the mean (95% CI) morning PEF (l/min) in smokers and non-smokers (-25 (95% CI -45 to -4), adjusted p = 0.019) and in the number of asthma exacerbations (6 v 1 in smokers and non smokers, respectively, p = 0.007). These differences were reduced between smokers and non-smokers receiving 2000 microg inhaled beclomethasone daily. CONCLUSIONS: Compared with non-smokers, smokers with mild persistent asthma are insensitive to the therapeutic effect of low dose inhaled corticosteroid treatment administered for a 12 week period. The disparity of the response between smokers and non smokers appears to be reduced with high dose inhaled corticosteroid. These findings have important implications for the management of individuals with mild asthma who smoke. PMID- 15790983 TI - Altered thoracic gas compression contributes to improvement in spirometry with lung volume reduction surgery. AB - BACKGROUND: Thoracic gas compression (TGC) exerts a negative effect on forced expiratory flow. Lung resistance, effort during a forced expiratory manoeuvre, and absolute lung volume influence TGC. Lung volume reduction surgery (LVRS) reduces lung resistance and absolute lung volume. LVRS may therefore reduce TGC, and such a reduction might explain in part the improvement in forced expiratory flow with the surgery. A study was conducted to determine the effect of LVRS on TGC and the extent to which reduced TGC contributed to an improvement in forced expiratory volume in 1 second (FEV1) following LVRS. METHODS: The effect of LVRS on TGC was studied using prospectively collected lung mechanics data from 27 subjects with severe emphysema. Several parameters including FEV1, expiratory and inspiratory lung resistance (Rle and Rli), and lung volumes were measured at baseline and 6 months after surgery. Effort during the forced manoeuvre was measured using transpulmonary pressure. A novel method was used to estimate FEV1 corrected for the effect of TGC. RESULTS: At baseline the FEV1 corrected for gas compression (NFEV1) was significantly higher than FEV1 (p<0.0001). FEV1 increased significantly from baseline (p<0.005) while NFEV1 did not change following surgery (p>0.15). TGC decreased significantly with LVRS (p<0.05). Rle and maximum transpulmonary pressure (TP(peak)) during the forced manoeuvre significantly predicted the reduction in TGC following the surgery (Rle: p<0.01; TP(peak): p<0.0001; adjusted R2 = 0.68). The improvement in FEV1 was associated with the reduction in TGC after surgery (p<0.0001, adjusted R2 = 0.58). CONCLUSIONS: LVRS decreased TGC by improving expiratory flow limitation. In turn, the reduction in TGC decreased its negative effect on expiratory flow and therefore explained, in part, the improvement in FEV1 with LVRS in this cohort. PMID- 15790984 TI - Oxidative stress and airway inflammation in severe exacerbations of COPD. AB - BACKGROUND: A study was undertaken to assess both oxidative stress and inflammation in the lungs of patients with chronic obstructive pulmonary disease (COPD) during severe and very severe exacerbations compared with those with stable COPD, healthy smokers, and non-smokers. Two sites within the lungs were compared: the large airways (in sputum) and the peripheral airways (by bronchoalveolar lavage (BAL)). METHODS: BAL fluid cell numbers and levels of tumour necrosis factor (TNFalpha) and interleukin (IL)-8 were measured as markers of airway inflammation and glutathione (GSH) levels as a marker of antioxidant status. Nuclear translocation of the pro-inflammatory transcription factors nuclear factor-kappaB (NF-kappaB) and activator protein 1 (AP-1) were also measured by electromobility shift assay in BAL fluid leucocytes and lung biopsy samples. RESULTS: Influx of inflammatory cells into the peripheral airways during exacerbations of COPD was confirmed. Increased IL-8 levels were detected in BAL fluid from patients with stable COPD compared with non-smokers and healthy smokers, with no further increase during exacerbations. In contrast, IL-8 levels in the large airways increased during exacerbations. GSH levels were increased in the BAL fluid of smokers (444%) and patients with stable COPD (235%) compared with non-smokers and were reduced during exacerbations (severe 89.2%; very severe 52.3% compared with stable COPD). NF-kappaB DNA binding in BAL leucocytes was decreased in healthy smokers compared with non-smokers (41.3%, n = 9, p<0.001) but did not differ in COPD patients, whereas AP-1 DNA binding was significantly decreased during exacerbations of COPD. CONCLUSION: There is evidence of increased oxidative stress in the airways of patients with COPD that is increased further in severe and very severe exacerbations of the disease. This is associated with increased neutrophil influx and IL-8 levels during exacerbations. PMID- 15790985 TI - Early onset of effect of salmeterol and fluticasone propionate in chronic obstructive pulmonary disease. AB - BACKGROUND: Combined treatment with inhaled corticosteroids and long acting beta2 agonists is approved for the treatment of chronic obstructive pulmonary disease (COPD), but little is known about the onset of effect of the combination. METHODS: Data were used from 1465 patients with COPD entered into a large 1 year double blind trial with daily measurements of peak expiratory flow (PEF) and symptom scores. RESULTS: PEF was significantly higher after 1 day in patients treated with salmeterol 50 microg twice daily or the salmeterol/fluticasone propionate combination 50/500 microg twice daily than placebo. In patients treated with fluticasone propionate 500 microg twice daily alone, PEF differed from placebo after 2 days. The differences after 2 weeks compared with placebo were 16 l/min (95% confidence interval (CI) 11 to 21), 11 l/min (95% CI 6 to 16), and 27 l/min (95% CI 22 to 33) for salmeterol, fluticasone propionate, and the salmeterol/fluticasone propionate combination, respectively. For all treatments the effect on PEF after 2 weeks was comparable to that seen at the end of the study. The difference between the salmeterol/fluticasone propionate combination and placebo after 2 weeks as a percentage of baseline was similar for PEF and clinic forced expiratory volume in 1 second (FEV1). Differences in breathlessness scores were statistically significant after 1 day for the group treated with salmeterol alone and after 2 days for the combination group. The 2 week change in FEV1 was only partly indicative of a long term response in individual patients. CONCLUSIONS: The effects of salmeterol and fluticasone propionate, alone or in combination, on PEF and breathlessness are seen within days and most of the obtainable effect on these parameters is reached within 2 weeks. PMID- 15790986 TI - Acute hypoxia simultaneously induces the expression of gp91phox and endothelial nitric oxide synthase in the porcine pulmonary artery. AB - BACKGROUND: The effect of hypoxia on the formation of superoxide (O2-), the expression of gp91phox and endothelial NO synthase (eNOS) were studied in pig intact pulmonary artery (PA) segments and PA vascular smooth muscle cells (PAVSMCs) and PA endothelial cells (PAECs). METHODS: Segments and cells were incubated under hypoxic conditions for 2 hours (with or without enzyme inhibitors) and the formation of O2- measured spectrophotometrically. Protein expression was assessed using Western blotting and immunocytochemistry. RESULTS: Hypoxia promoted the formation of O2- in PA segments, PAVSMCs and PAECs, an effect inhibited by diphenylene iodonium and apocynin (NAD[P]H oxidase inhibitors). Hypoxia induced O2- formation was enhanced by inhibition of eNOS and augmented by endotoxin and cytokines and re-oxygenation. Hypoxia also promoted the expression of gp91phox and eNOS. In intact PA segments hypoxia induced the expression of nitrotyrosine and eNOS in the endothelium. CONCLUSIONS: The simultaneous upregulation of NAD[P]H oxidase and eNOS in response to hypoxia in the PA results in the simultaneous formation of O2-, NO, and ONOO-. This may represent either a protective mechanism designed to counter the pro-oxidant effect of hypoxia or a novel pathological mechanism underlying the progression of acute respiratory distress syndrome (ARDS). PMID- 15790987 TI - Is late diagnosis of lung cancer inevitable? Interview study of patients' recollections of symptoms before diagnosis. AB - BACKGROUND: A study was undertaken to explore the pathway to diagnosis among a group of patients recently diagnosed with lung cancer. METHODS: A directed interview study triangulating patients' accounts with hospital and GP records was performed with 22 men and women recently diagnosed with lung cancer at two cancer centres in the south and north of England. The main outcome measures were the symptoms leading up to a diagnosis of lung cancer and patient and GP responses before diagnosis. RESULTS: Patients recalled having new symptoms for many months, typically over the year before their diagnosis, irrespective of their disease stage once diagnosed. Chest symptoms (cough, breathing changes, and pain in the chest) were common, as were systemic symptoms (fatigue/lethargy, weight loss and eating changes). Although symptoms were reported as being marked changes in health, these were not in the main (with the exception of haemoptysis) interpreted as serious by patients at the time and not acted on. Once the trigger for action occurred (the event that took patients to their GP or elsewhere in the healthcare system), events were relatively speedy and were faster for patients who presented via their GP than via other routes. Patients' beliefs about health changes that may indicate lung cancer appeared to have played a part in delay in diagnosis. CONCLUSION: Further investigation of the factors influencing the timing of diagnosis in lung cancer is warranted since it appears that patients did not readily attend GP surgeries with symptoms. Insight into patients' perspectives on their experience before diagnosis may help medical carers to recognise patients with lung cancer more easily so that they can refer them for diagnosis and treatment. Encouragement to present early with signs of lung cancer should be considered alongside other efforts to speed up diagnosis and treatment. PMID- 15790988 TI - Association of tumour necrosis factor alpha variants with the CF pulmonary phenotype. AB - BACKGROUND: The pulmonary phenotype in patients with cystic fibrosis (CF), even in those with the same CF transmembrane conductance regulator (CFTR) genotype, is variable and must therefore be influenced by secondary genetic factors as well as environmental factors. Possible candidate genes that modulate the CF lung phenotype may include proinflammatory cytokines. One such protein is tumour necrosis factor alpha (TNFalpha), a member of the immune system. METHODS: Three polymorphic loci in the promoter (-851c/t, -308g/a, -238g/a) and one polymorphic locus in intron 1 (+691g ins/del) of the TNFalpha gene were typed by a single nucleotide primer extension assay in CF patients and healthy controls. Spirometric data and first age of infection with Pseudomonas aeruginosa were collected retrospectively from patients' medical records. RESULTS: An association was found between the TNFalpha +691g ins/del polymorphic locus and severity of CF lung disease. Patients heterozygous for +691g ins and +691g del were more likely to have better pulmonary function (mean (SD) forced expiratory volume in 1 second (FEV1) 79.7 (12.8)% predicted) than patients homozygous for +691g ins (mean (SD) FEV1 67.5 (23.0)% predicted; p = 0.008, mean difference 12.2%, 95% CI 3.5 to 21.0). Also, patients heterozygous for +691g ins and +691g del were more likely to have an older first age of infection with P aeruginosa (mean (SD) 11.4 (6.0) years) than patients homozygous for +691g ins (mean (SD) 8.3 (4.6) years; p = 0.018, mean difference 3.1 years, 95% CI 0.5 to 5.6). An association was also found with the -851c/t polymorphic locus. In the group of patients with more severe FEV1% predicted, a higher proportion of patients were homozygous for the 851c allele than in the other group of patients (p = 0.04, likelihood ratio chi2, odds ratio = 2.4). CONCLUSION: TNFalpha polymorphisms are associated with the severity of CF lung disease in Czech and Belgian patients with CF. PMID- 15790989 TI - A survey of sexual and reproductive health in men with cystic fibrosis: new challenges for adolescent and adult services. AB - BACKGROUND: Improved survival dramatically alters the consequences of adult co morbidities in men with cystic fibrosis (CF) such as male infertility. Few studies have systematically addressed the impact of sexual and reproductive health issues in these men or considered the implications for healthcare delivery. METHOD: A descriptive cohort study was undertaken using a sexual and reproductive health survey of men from a large adult CF centre, including men with lung transplantation. RESULTS: The mean (SD) age of the 94 men (response rate 75%) was 30.5 (7.6) years. 94% knew that men with CF had reduced fertility. Men first heard about infertility later than desired (p<0.001) and only 53% heard from their preferred source. Men who were told about infertility when older were more likely to be upset than those told earlier (p<0.01). 53% of men had undergone semen analysis: 68% of men who had not been tested wanted semen analysis. 73% believed semen analysis should occur before 18, but the youngest age of testing was 24 years. In adolescence, one in three men had assumed they did not need to use condoms and one in 10 had confused infertility with impotence. 66% of men wanted more information on reproductive options and 84% wanted children. Seventeen men were parents by natural conception (n = 1), micro epididymal sperm aspiration (n = 6), donor sperm (n = 9), and through step children (n = 1). CONCLUSIONS: Men with CF desire more sexual and reproductive health information. Earlier discussion of sexual and reproductive health is indicated in paediatric settings, and semen analysis should be routinely offered. In adult services greater discussion of reproductive health options is indicated. PMID- 15790990 TI - Respiratory symptoms in older people and their association with mortality. AB - BACKGROUND: A study was undertaken to assess the prevalence of reported respiratory symptoms in a population aged over 75 years and their association with mortality. METHODS: A total of 14 458 people aged 75 years and over participating in a trial of health screening of older people in general practice answered questions on three respiratory symptoms: cough, sputum production, and wheeze. The association of symptoms with mortality was examined for all cause and respiratory causes of death taking account of potential confounders. RESULTS: Coughing up phlegm in winter mornings had a prevalence of 27.0% (95% confidence interval (CI) 26.8 to 27.2). Those with this symptom had an adjusted hazard ratio for all cause mortality of 1.35 (95% CI 1.21 to 1.50), p<0.001 and for respiratory specific mortality of 2.01 (95% CI 1.66 to 2.41), p<0.001. Phlegm at any time of the day in winter had a prevalence of 16.5% (95% CI 16.3 to 16.7) with hazard ratios for all cause and respiratory specific mortality of 1.28 (95% CI 1.15 to 1.42) and 2.28 (95% CI 1.92 to 2.70), p<0.001. Wheeze or whistling from the chest had a prevalence of 14.3% (95% CI 14.1 to 14.5) with hazard ratios of 1.45 (95% CI 1.31 to 1.61) and 2.86 (95% CI 2.45 to 3.35), p<0.001. CONCLUSIONS: The prevalence of respiratory symptoms is widespread among elderly people and their presence is a strong predictor of mortality. PMID- 15790993 TI - Pulmonary amoebiasis presenting as superior vena cava syndrome. AB - Pulmonary amoebiasis without liver involvement occurs sporadically as a result of haematogenous spread from a primary site, the colon. The case history is presented of a patient who developed superior vena cava syndrome due to a pulmonary amoebic abscess without liver involvement. He was initially suspected of having a neoplasm but a combination of tests including histological examination of the H&E stained excised tissue, immunofluorescence using anti Entamoeba histolytica antibodies, and serology confirmed the diagnosis of amoebiasis. To our knowledge this is the first description of pulmonary amoebiasis presenting as superior vena cava syndrome. PMID- 15790991 TI - Assessment of progression of COPD: report of a workshop held in Leuven, 11-12 March 2004. AB - Recently performed long term trials have enhanced the insight into the assessment of progression of COPD. The present review focuses on the initial assessment of COPD in general practice and the assessment of disease progression. Several variables may be used to assess this progression, all of which are associated with significant methodological problems. Finding the appropriate mix of outcome measures to capture all aspects of disease progression is a significant challenge. PMID- 15790992 TI - Effect of treatments on the progression of COPD: report of a workshop held in Leuven, 11-12 March 2004. AB - During the last decade several long term studies of interventions in patients with COPD have been published. This review analyses the potential of these interventions to alter the progression of the condition. The only treatment that has unequivocally been shown to reduce the rate of decline in FEV(1) is smoking cessation. Active psychological intervention in combination with pharmacotherapy is required. Other treatments may have an effect on the rate of decline in FEV(1) but this appears to be very small, at most. Several treatments affect the exacerbation rate and therefore might affect the progression of the disease. Further studies are warranted to examine this effect. PMID- 15790994 TI - VEGF in idiopathic ILD. PMID- 15790995 TI - CARD 15 gene mutations in sarcoidosis. PMID- 15790996 TI - Infected pulmonary sequestration caused by Mycobacterium kansasii. PMID- 15790997 TI - Pulmonary adenocarcinoma presenting as a giant pleural mass. PMID- 15790999 TI - N-coumaroyldopamine and N-caffeoyldopamine increase cAMP via beta 2-adrenoceptors in myelocytic U937 cells. AB - N-caffeoyldopamine is a phytochemical found in various plants, including cocoa (Theobroma cacao L.). N-caffeoyldopamine and its natural analogs (N cinnamoyldopamine, N-coumaroyldopamine, N-feruloyldopamine, and N sinapoyldopamine) were synthesized and investigated to determine their potency as beta-adrenoceptor agonists, because they have chemical structural moieties found in beta-adrenoceptor agonists. Among the compounds tested in this study, N coumaroyldopamine and N-caffeoyldopamine were the two most potent compounds, able to increase cAMP at the concentrations < 0.05 microM in U937 cells. The decreasing order of potency was N-coumaroyldopamine > N-caffeoyldopamine > N feruloyldopamine > N-sinapoyldopamine > N-cinnamoyldopamine. Using beta2-specific antagonists (butoxamine and ICI 118551), N-coumaroyldopamine and N caffeoyldopamine were found to increase cAMP via beta2-adrenoceptors in U937 cells. In producing cAMP in U937 cells, N-coumaroyldopamine and N caffeoyldopamine were as potent as several well-known beta2-adrenoceptor agonists (salbutamol, procaterol, and fenoterol). These results indicate that N coumaroyldopamine and N-caffeoyldopamine are potent compounds able to increase cAMP via beta2-adrenoceptors in U937 cells, and may have potential effects on human health. PMID- 15790998 TI - Femtomolar concentrations of dextromethorphan protect mesencephalic dopaminergic neurons from inflammatory damage. AB - Inflammation in the brain has increasingly been recognized to play an important role in the pathogenesis of several neurodegenerative disorders, including Parkinson's disease (PD). Progress in the search for effective therapeutic strategies that can halt this degenerative process remains limited. We previously showed that micromolar concentrations of dextromethorphan (DM), a major ingredient of widely used antitussive remedies, reduced the inflammation-mediated degeneration of dopaminergic neurons through the inhibition of microglial activation. In this study, we report that femto- and micromolar concentrations of DM (both pre- and post-treatment) showed equal efficacy in protecting lipopolysaccharide (LPS) -induced dopaminergic neuron death in midbrain neuron glia cultures. Both concentrations of DM decreased LPS-induced release of nitric oxide, tumor necrosis factor-alpha, prostaglandin E2 and superoxide from microglia in comparable degrees. The important role of superoxide was demonstrated by DM's failure to show a neuroprotective effect in neuron-glia cultures from NADPH oxidase-deficient mice. These results suggest that the neuroprotective effect elicited by femtomolar concentrations of DM is mediated through the inhibition of LPS-induced proinflammatory factors, especially superoxide. These findings suggest a novel therapeutic concept of using "ultra low" drug concentrations for the intervention of inflammation-related neurodegenerative diseases. PMID- 15791000 TI - Regulation of the human atrial myosin light chain 1 promoter by Ca2+-calmodulin dependent signaling pathways. AB - We investigated expression regulation of the human atrial myosin light chain 1 (hALC-1) gene using a cardiomyocyte H9c2 cell line stably transfected with a construct consisting of the human ALC-1 promoter cloned in front of the luciferase gene (H9c2T1). H9c2T1 cells were stimulated with vasopressin, which is known to induce cardiomyocyte hypertrophy and to activate a panel of signaling pathways. Those pathways involved in hALC-1 promoter activity regulation were dissected by using pharmacological inhibitor substances. Stimulation with vasopressin was associated with nuclear NFAT translocation and significantly increased human ALC-1 promoter activity. Inhibition of calcineurin by cyclosporin A blocked the effects of vasopressin on ALC-1 promoter activity to approximately 50%. This suggests that the Ca2+-calmodulin-calcineurin-NFAT pathway is involved in human ALC-1 promoter activation. However, inhibition of multifunctional Ca2+ calmodulin-dependent protein kinases (CaMK) by KN-93 decreased human ALC-1 promoter activity to almost basal levels. CaMK regulation of ALC-1 promoter activity effect could well be mediated by CaMKIV, which accumulated in the nucleus upon vasopressin stimulation. Inhibition of protein kinase C (PKC) isoforms by bisindolylmaleimide had no significant influence on human ALC-1 promoter activity. Thus, our results demonstrate a dominant role of Ca2+ calmodulin-dependent signaling pathways in the regulation of human ALC-1 expression. PMID- 15791001 TI - Platelet-activating factor increases VE-cadherin tyrosine phosphorylation in mouse endothelial cells and its association with the PtdIns3'-kinase. AB - Platelet-activating factor (PAF), a potent inflammatory mediator, is involved in endothelial permeability. This study was designed to characterize PAF receptor (PAF-R) expression and its specific contribution to the modifications of adherens junctions in mouse endothelial cells. We demonstrated that PAF-R was expressed in mouse endothelial cells and was functionally active in stimulating p42/p44 MAPK and phosphatidylinositol 3-kinase (PtdIns3'-kinase)/Akt activities. Treatment of cells with PAF induced a rapid time- and dose-dependent (10(-7) to 10(-10) M) increase in tyrosine phosphorylation of a subset of proteins ranging from 90 to 220 kDa, including the VE-cadherin, the latter effect being prevented by the tyrosine kinase inhibitors herbimycin A and bis-tyrphostin. We demonstrated that PAF promoted formation of multimeric aggregates of VE-cadherin with PtdIns3' kinase, which was also inhibited by herbimycin and bis-tyrphostin. Finally, we show by immunostaining of endothelial cells VE-cadherin that PAF dissociated adherens junctions. The present data provide the first evidence that treatment of endothelial cells with PAF promoted activation of tyrosine kinases and the VE cadherin tyrosine phosphorylation and PtdIns3'-kinase association, which ultimately lead to the dissociation of adherens junctions. Physical association between PtdIns3'-kinase, serving as a docking protein, and VE-cadherin may thus provide an efficient mechanism for amplification and perpetuation of PAF-induced cellular activation. PMID- 15791002 TI - Expression of the Leo1-like domain of replicative senescence down-regulated Leo1 like (RDL) protein promotes senescence of 2BS fibroblasts. AB - Replicative senescence is thought to relate to aging in vivo and tumor suppression. In this report, we isolated a gene and designated it as RDL (replicative senescence down-regulated Leo1-like gene). RDL's expression decreased upon replicative senescence of human diploid 2BS fibroblasts. Overexpression of RDL slightly delayed 2BS fibroblast senescence, whereas suppression of RDL expression imposed no obvious effects on senescence. However, introduction of cDNA fragment encoding the Leo1-like domain of RDLp (Leo) alone shortened the replicative life span of 2BS fibroblasts and promoted several senescent features; the introduction of truncated RDL cDNA fragment resulting from deletion of Leo (RDL-Leo-) significantly prolonged 2BS life span and caused a noticeable delay of these senescent features. We demonstrated that introduction of Leo obviously increased the expression of p16INK4a, p21WAF1, and PTEN, whereas introduction of RDL-Leo- distinctly decreased p16INK4a expression. Taken together, our results suggest that the Leo1-like domain of RDLp is a senescence associated domain that accelerates the senescence of 2BS fibroblasts and that there should be another counteractive domain in the remaining part of RDLp. PMID- 15791003 TI - Aggregated alpha-synuclein activates microglia: a process leading to disease progression in Parkinson's disease. AB - A growing body of evidence indicates that an inflammatory process in the substantia nigra, characterized by activation of resident microglia, likely either initiates or aggravates nigral neurodegeneration in Parkinson's disease (PD). To study the mechanisms by which nigral microglia are activated in PD, the potential role of alpha-synuclein (a major component of Lewy bodies that can cause neurodegeneration when aggregated) in microglial activation was investigated. The results demonstrated that in a primary mesencephalic neuron glia culture system, extracellular aggregated human alpha-synuclein indeed activated microglia; microglial activation enhanced dopaminergic neurodegeneration induced by aggregated alpha-synuclein. Furthermore, microglial enhancement of alpha-synuclein-mediated neurotoxicity depended on phagocytosis of alpha-synuclein and activation of NADPH oxidase with production of reactive oxygen species. These results suggest that nigral neuronal damage, regardless of etiology, may release aggregated alpha-synuclein into substantia nigra, which activates microglia with production of proinflammatory mediators, thereby leading to persistent and progressive nigral neurodegeneration in PD. Finally, NADPH oxidase could be an ideal target for potential pharmaceutical intervention, given that it plays a critical role in alpha-synuclein-mediated microglial activation and associated neurotoxicity. PMID- 15791004 TI - Myostatin propeptide-mediated amelioration of dystrophic pathophysiology. AB - Mutations in myostatin (GDF8) cause marked increases in muscle mass, suggesting that this transforming growth factor-beta (TGF-beta) superfamily member negatively regulates muscle growth. Myostatin blockade therefore offers a strategy for reversing muscle wasting in Duchenne's muscular dystrophy (DMD) without resorting to genetic manipulation. Here, we demonstrate that pharmacological blockade using a myostatin propeptide stabilized by fusion to IgG Fc improved pathophysiology of the mdx mouse model of DMD. Functional benefits evidenced by specific force improvement, exceeded those reported previously using myostatin antibody-mediated blockade. More importantly, use of a propeptide blockade strategy obviates possibilities of anti-idiotypic responses that could potentially limit the effectiveness of antibody-mediated myostatin blockade strategies over time. This study provides a novel pharmacological strategy for treatment of diseases associated with muscle wasting such as DMD and since it uses an endogenous inhibitor of myostatin should help circumvent technical hurdles and toxicity associated with conventional gene or cell based therapies. PMID- 15791005 TI - Microglial NADPH oxidase is a novel target for femtomolar neuroprotection against oxidative stress. AB - Inflammation has been increasingly recognized to contribute to the pathogenesis of Parkinson's disease. Several compounds are neuroprotective at femtomolar concentrations through the inhibition of inflammation. However, the mechanisms mediating femtomolar-acting compounds are poorly understood. Here we show that both gly-gly-phe (GGF), a tri-peptide contained in the dynorphin opioid peptide, and naloxone are neuroprotective at femtomolar concentrations against LPS-induced dopaminergic neurotoxicity through the reduction of microglial activation. Mechanistic studies demonstrated the critical role of NADPH oxidase in the GGF and naloxone inhibition of microglial activation and associated DA neurotoxicity. Pharmacophore analysis of the neuroprotective dynorphin peptides and naloxone revealed common chemical properties (hydrogen bond acceptor, hydrogen bond donor, positive ionizable, hydrophobic) of these femtomolar-acting compounds. These results support a common high-affinity site of action for several femtomolar acting compounds, where NADPH oxidase is the critical mechanism governing neuroprotection, suggesting a novel avenue of anti-inflammatory and neuroprotective therapy. PMID- 15791006 TI - PARG activity mediates intestinal injury induced by splanchnic artery occlusion and reperfusion. AB - Poly (ADP-ribosyl)ation, an early post-translational modification in response to DNA damage, is catalyzed by poly (ADP-ribose) polymerase (PARP-1) and catabolized by poly(ADP-ribose) glycohydrolase (PARG). The aim of this study was to investigate the role of PARG on the modulation of the inflammatory response caused by splanchnic ischemia and reperfusion. SAO shock in rats and wild-type (WT) mice was associated with a significant neutrophil infiltration in the ileum and production of tumor necrosis factor-alpha (TNF-alpha). Reperfused ileum tissue sections from SAO-shocked WT mice and rats showed positive staining for P selectin and ICAM-1 localized mainly in the vascular endothelial cells. Genetic disruption of the PARG gene in mice or pharmacological inhibition of PARG by PARG inhibitors significantly improved the histological status of the reperfused tissues associated with reduced expression of P-selectin and ICAM-1, neutrophil infiltration into the reperfused intestine, and TNF-alpha production. These results suggest that PARG activity modulates the inflammatory response in ischemia/reperfusion and participates in end (target) organ damage under these conditions. PMID- 15791007 TI - Structural basis for high-affinity volatile anesthetic binding in a natural 4 helix bundle protein. AB - Physiologic sites for inhaled anesthetics are presumed to be cavities within transmembrane 4-alpha-helix bundles of neurotransmitter receptors, but confirmation of binding and structural detail of such sites remains elusive. To provide such detail, we screened soluble proteins containing this structural motif, and found only one that exhibited evidence of strong anesthetic binding. Ferritin is a 24-mer of 4-alpha-helix bundles; both halothane and isoflurane bind with K(A) values of approximately 10(5) M(-1), higher than any previously reported inhaled anesthetic-protein interaction. The crystal structures of the halothane/apoferritin and isoflurane/apoferritin complexes were determined at 1.75 A resolution, revealing a common anesthetic binding pocket within an interhelical dimerization interface. The high affinity is explained by several weak polar contacts and an optimal host/guest packing relationship. Neither the acidic protons nor ether oxygen of the anesthetics contribute to the binding interaction. Compared with unliganded apoferritin, the anesthetic produced no detectable alteration of structure or B factors. The remarkably high affinity of the anesthetic/apoferritin complex implies greater selectivity of protein sites than previously thought, and suggests that direct protein actions may underlie effects at lower than surgical levels of anesthetic, including loss of awareness. PMID- 15791008 TI - Thrombolytic (fibrinolytic) drugs and progress in treating cardiovascular disease. PMID- 15791009 TI - Smoke-free bars in Ireland: a runaway success. PMID- 15791010 TI - Australia: epidemiology classes, shredding, and calls to "the garbageman": a day in the life of tobacco folk. PMID- 15791016 TI - How do minimum cigarette price laws affect cigarette prices at the retail level? AB - OBJECTIVES: Half of US states have minimum cigarette price laws that were originally passed to protect small independent retailers from unfair price competition with larger retailers. These laws prohibit cigarettes from being sold below a minimum price that is set by a formula. Many of these laws allow cigarette company promotional incentives offered to retailers, such as buydowns and master-type programmes, to be calculated into the formula. Allowing this provision has the potential to lower the allowable minimum price. This study assesses whether stores in states with minimum price laws have higher cigarette prices and lower rates of retailer participation in cigarette company promotional incentive programmes. DESIGN: Retail cigarette prices and retailer participation in cigarette company incentive programmes in 2001 were compared in eight states with minimum price laws and seven states without them. New York State had the most stringent minimum price law at the time of the study because it excluded promotional incentive programmes in its price setting formula; cigarette prices in New York were compared to all other states included in the study. RESULTS: Cigarette prices were not significantly different in our sample of US states with and without cigarette minimum price laws. Cigarette prices were significantly higher in New York stores than in the 14 other states combined. CONCLUSIONS: Most existing minimum cigarette price laws appear to have little impact on the retail price of cigarettes. This may be because they allow the use of promotional programmes, which are used by manufacturers to reduce cigarette prices. New York's strategy to disallow these types of incentive programmes may result in higher minimum cigarette prices, and should also be explored as a potential policy strategy to control cigarette company marketing practices in stores. Strict cigarette minimum price laws may have the potential to reduce cigarette consumption by decreasing demand through increased cigarette prices and reduced promotional activities at retail outlets. PMID- 15791017 TI - Higher cigarette prices influence cigarette purchase patterns. AB - OBJECTIVE: To examine cigarette purchasing patterns of current smokers and to determine the effects of cigarette price on use of cheaper sources, discount/generic cigarettes, and coupons. BACKGROUND: Higher cigarette prices result in decreased cigarette consumption, but price sensitive smokers may seek lower priced or tax-free cigarette sources, especially if they are readily available. This price avoidance behaviour costs states excise tax money and dampens the health impact of higher cigarette prices. METHODS: Telephone survey data from 3602 US smokers who were originally in the COMMIT (community intervention trial for smoking cessation) study were analysed to assess cigarette purchase patterns, use of discount/generic cigarettes, and use of coupons. RESULTS: 59% reported engaging in a high price avoidance strategy, including 34% who regularly purchase from a low or untaxed venue, 28% who smoke a discount/generic cigarette brand, and 18% who report using cigarette coupons more frequently that they did five years ago. The report of engaging in a price avoidance strategy was associated with living within 40 miles of a state or Indian reservation with lower cigarette excise taxes, higher average cigarette consumption, white, non-Hispanic race/ethnicity, and female sex. CONCLUSION: Data from this study indicate that most smokers are price sensitive and seek out measures to purchase less expensive cigarettes, which may decrease future cessation efforts. PMID- 15791018 TI - The federal initiative to halt the sale of tobacco to children--the Synar Amendment, 1992-2000: lessons learned. AB - BACKGROUND: The Synar Amendment was enacted by the US Congress in 1992 to require states and territories to establish and enforce laws prohibiting the sale of tobacco to minors. OBJECTIVE: To describe state and federal efforts to comply with the Synar mandate. METHODS: State and federal actions were examined for the eight years following enactment. DATA SOURCES: Federal documents from 1992-2003, annual block grant applications from 59 states and territories describing activities during federal fiscal years 1995-2000. MEASURES: Whether applicants made a good faith effort to comply by enacting a law, enforcing it with inspections and penalties, conducting a valid survey and meeting violation rate targets set by the Department of Health and Human Services (DHHS). RESULTS: Between 1996 and 2000, 26 states had made a good faith effort to comply with Synar every year. In 2000, 57 jurisdictions (excluding Maryland and Montana) had established laws without loopholes, 57 conducted a valid survey, and 54 actively enforced their laws. By 2002, violation rates had dropped substantially everywhere but Alaska and a few small territories. No state reached the violation rate goal of 20% without penalising violators. CONCLUSIONS: The Synar Amendment has resulted in the universal adoption of laws prohibiting tobacco sales to minors and almost universal enforcement of those laws, resulting in dramatically reduced violation rates. Implementation was slowed significantly by a lack of good faith effort in many states and by DHHS's decision not to require states to enforce their laws by penalising lawbreakers. PMID- 15791019 TI - If smoking increases absences, does quitting reduce them? AB - OBJECTIVE: This study examined the impact of smoking, quitting, and time since quit on absences from work. METHODS: Data from the nationally representative Tobacco Use Supplements of the 1992/93, 1995/96, and 1998/99 Current Population Surveys were used. The study included full time workers aged between 18-64 years, yielding a sample size of 383 778 workers. A binary indicator of absence due to sickness in the last week was analysed as a function of smoking status including time since quit for former smokers. Extensive demographic variables were included as controls in all models. RESULTS: In initial comparisons between current and former smokers, smoking increased absences, but quitting did not reduce them. However, when length of time since quit was examined, it was discovered that those who quit within the last year, and especially the last three months, had a much greater probability of absences than did current smokers. As the time since quitting increased, absences returned to a rate somewhere between that of never and current smokers. Interactions between health and smoking status significantly improved the fit of the model. CONCLUSIONS: Smokers who quit reduced their absences over time but increase their absences immediately after quitting. Quitting ill may account for some but not all of this short run impact. PMID- 15791020 TI - Educational inequalities in smoking among men and women aged 16 years and older in 11 European countries. AB - OBJECTIVE: To determine those groups who are at increased risk of smoking related diseases, we assessed in which male and female generations smoking was more prevalent among lower educated groups than among the higher educated, in 11 European countries. DESIGN: Cross sectional analysis of data on smoking, covering the year 1998, from a social survey designed for all member states of the European Union. SUBJECTS: Higher and lower educated men and women aged 16 years and older from 11 member states of the European Union. OUTCOME MEASURES: Age standardised prevalence rates by education and prevalence odds ratios of current and ever daily smoking comparing lower educated groups with higher educated groups. RESULTS: A north-south gradient in educational inequalities in current and ever daily smoking was observed for women older than 24 years, showing larger inequalities in the northern countries. Such a gradient was not observed for men. A disadvantage for the lower educated in terms of smoking generally occurred later among women than among men. Indications of inequalities in smoking in the age group 16-24 years were observed for all countries, with the exception of women from Greece and Portugal. CONCLUSIONS: Preventing and reducing smoking among lower educated subgroups should be a priority of policies aiming to reduce inequalities in health in Europe. If steps are not taken to control tobacco use among the lower educated groups specifically, inequalities in lung cancer and other smoking related diseases should be anticipated in all populations of the European Union, and both sexes. PMID- 15791021 TI - Validity of self reports in a cohort of Swedish adolescent smokers and smokeless tobacco (snus) users. AB - OBJECTIVE: To validate self reports of cigarette and smokeless tobacco (snus) use in a prospective cohort of adolescents. DESIGN: A cross sectional analysis of a cohort sub-sample. SETTING: County of Stockholm, Sweden. SUBJECTS: 520 adolescents in the final grade of junior high school (mean age 15.0 years). MAIN OUTCOME MEASURE: Concordance between self reported tobacco use and saliva cotinine concentration. RESULTS: Using a cut point of 5 ng/ml saliva cotinine to discriminate active tobacco use, there was a 98% concordance between self reported non-use in the past month and cotinine concentration. The sensitivity of the questionnaire compared to the saliva cotinine test, used as the gold standard, was 90% and the specificity 93%. One hundred and fifteen out of 520 subjects (22%) reported monthly tobacco use. Among these, 67% (46/69) of the exclusive cigarette smokers, 82% (23/28) of exclusive snus users, and 94% (15/16) of mixed users (cigarettes + snus) had cotinine concentrations above 5 ng/ml. Among subjects reporting daily use 96% (64/67) had saliva cotinine concentrations above the cut point. Exclusive current cigarette users were more likely to be classified discordantly by questionnaire and cotinine test compared to snus users (odds ratio 3.2, 95% confidence interval 1.2 to 8.6). CONCLUSION: This study confirms the reliability of adolescents' self reported tobacco use. In a context of low exposure to environmental tobacco smoke a cut off for saliva cotinine of 5 ng/ml reliably discriminated tobacco users from non-users. Irregular use of tobacco in this age group probably explains the discrepancy between self reported use and cotinine concentrations. PMID- 15791022 TI - The limits of competing interest disclosures. AB - OBJECTIVE: To assess the effectiveness of conflict of interest disclosure policies by comparing a competing interests disclosure statement that met the requirements established by the journal in a 2003 article on health effects of secondhand smoke based on the American Cancer Society CPS-I dataset with internal tobacco industry documents describing financial ties between the tobacco industry and authors of the study. DESIGN: Descriptive analysis of internal tobacco industry documents retrieved from the Legacy Tobacco Documents Library, University of California, San Francisco. RESULTS: Meeting the requirements for financial disclosure established by the journal did not provide the reader with a full picture of the tobacco industry's involvement with the study authors. The tobacco industry documents reveal that the authors had long standing financial and other working relationships with the tobacco industry. CONCLUSION: These findings are another example of how simply requiring authors to disclose financial ties with the tobacco industry may not be adequate to give readers (and reviewers) a full picture of the author's relationship with the tobacco industry. The documents also reveal that the industry funds research to enhance its credibility and endeavours to work with respected scientists to advance its goals. These findings question the adequacy of current journal policies regarding competing interest disclosures and the acceptability of tobacco industry funding for academic research. PMID- 15791023 TI - Emotions for sale: cigarette advertising and women's psychosocial needs. AB - OBJECTIVE: To explore messages of psychosocial needs satisfaction in cigarette advertising targeting women and implications for tobacco control policy. METHODS: Analysis of internal tobacco industry documents and public advertising collections. RESULTS: Tobacco industry market research attempted to identify the psychosocial needs of different groups of women, and cigarette advertising campaigns for brands that women smoke explicitly aimed to position cigarettes as capable of satisfying these needs. Such positioning can be accomplished with advertising that downplays or excludes smoking imagery. As women's needs change with age and over time, advertisements were developed to reflect the needs encountered at different stages in women's lives. Cigarette brands for younger women stressed female camaraderie, self confidence, freedom, and independence; cigarette brands for older women addressed needs for pleasure, relaxation, social acceptability, and escape from daily stresses. CONCLUSIONS: Psychosocial needs satisfaction can be communicated without reference to cigarettes or smoking. This may explain why partial advertising bans are ineffective and comprehensive bans on all forms of tobacco marketing are effective. Counter-advertising should attempt to expose and undermine the needs satisfaction messages of cigarette advertising campaigns directed at women. PMID- 15791024 TI - Tobacco on campus: industry marketing and tobacco control policy among post secondary institutions in Canada. AB - OBJECTIVE: Post-secondary institutions provide a unique opportunity to implement and evaluate leading edge tobacco policies, while influencing a key group of young adults. To date, however, we know little about the tobacco control environment at post-secondary institutions outside the USA. DESIGN: Telephone surveys were conducted with campus informants from 35 post-secondary institutions in Canada to evaluate tobacco control policies and the presence of tobacco marketing on campus. MAIN OUTCOME MEASURES: Tobacco marketing on campus, tobacco control policies, and attitudes towards tobacco control. RESULTS: The findings indicate that tobacco marketing is prevalent among post-secondary institutions in Canada. Every university and half of all colleges surveyed had participated in some form of tobacco marketing in the past year. Among universities, 80% had run a tobacco advertisement in their paper and 18% had hosted a tobacco sponsored nightclub event. Tobacco control policies varied considerably between institutions. Although several campuses had introduced leading edge policies, such as campus wide outdoor smoking restrictions and tobacco sales bans, there is a general lack of awareness of tobacco issues among campus decision makers and fundamental public health measures, such as indoor smoke-free policies, have yet to be introduced in many cases. CONCLUSIONS: Post-secondary institutions in Canada remain tobacco friendly environments. Without increased direction and support from the public health community, post-secondary institutions will continue to lag behind, rather than lead current policy standards. PMID- 15791025 TI - "Pack year" smoking histories: what about patients who use loose tobacco? AB - Life time smoking of ready made cigarettes can be readily quantified as "pack years" smoked, but methods for measuring loose tobacco use are less well established. In this study the frequency of loose tobacco use by 247 hospital in patients was determined; 64% were current or ex-smokers, 41.3% of whom (25.9% of participants) had smoked loose tobacco. A formula was developed for converting loose tobacco use to pack years smoked, based on the weight of tobacco in ready made cigarettes; 12.5 g or half an ounce of loose tobacco was approximately equivalent to one packet of 20 cigarettes. Using a questionnaire it was found that hospital physicians of all grades were able to convert smoking histories of ready made cigarettes, but not loose tobacco, into number of "pack years" smoked. PMID- 15791026 TI - The case for smoker-free workplaces. PMID- 15791027 TI - The smoker-free workplace: the case against. PMID- 15791028 TI - Effects of glucose abnormalities on in-hospital outcome after coronary intervention for acute myocardial infarction. AB - BACKGROUND: The effects of glucose abnormalities on outcomes after percutaneous coronary intervention (PCI) remain unclear. We examined the association between glucose abnormalities and in-hospital outcome in patients undergoing PCI for acute myocardial infarction (AMI). METHODS AND RESULTS: A total of 849 patients with AMI who were admitted within 12 h after symptom onset and underwent emergency PCI were classified according to the presence or absence of admission hyperglycemia, defined as a blood glucose level on admission of >11 mmol/L and whether they had a history of diabetes mellitus: group 1 (n = 504), non-diabetic patients without admission hyperglycemia; group 2 (n = 111), diabetic patients without admission hyperglycemia; group 3 (n = 87), non-diabetic patients with admission hyperglycemia; and group 4 (n = 147), diabetic patients with admission hyperglycemia. Among groups 1, 2, 3 and 4, in-hospital mortality was 2.6, 2.7, 11.5 and 8.8%, respectively (p < 0.01). Multivariate analysis showed that compared with group 1 patients, the odds ratio (95%confidence interval) for in hospital mortality among those in groups 2, 3, and 4 were 0.80 (0.24-2.60, p = 0.708), 2.29 (1.10-5.49, p = 0.039), and 2.14 (1.14-4.69, p = 0.048), respectively. CONCLUSIONS: In-patients undergoing PCI for AMI, admission hyperglycemia, irrespective of the presence or absence of diabetes, is associated with increased in-hospital mortality, whereas diabetes without admission hyperglycemia is not. PMID- 15791029 TI - Late restenosis of the balloon-dilated site: serial angiographic observations beyond 7 years. AB - BACKGROUND: The present retrospective study was performed to assess the long-term (>7 years) fate of stabilized balloon-dilated sites. METHODS AND RESULTS: Between February and April 1986, 171 patients underwent successful percutaneous balloon angioplasty. Early restenosis (<1 year) occurred in 53%, but repeat balloon angioplasty stabilized the balloon-dilated site. The early period was defined as 6 months, late years as 3-5 years and long-term years as 7-12 years. Angiographic evaluation at both early year or late year periods (mean = 4.7 years) and long term (mean = 10.4 years) periods following stabilization was available in 71 patients (94 lesions) with mean age of 61.7+/-8.5 years. Of the 71 patients 69.6% were male. Restenosis occurring after 1 year was defined as late restenosis. The mean diameter stenosis changed from 6 months (50.3+/-12.4%) to late-period (44.2 +/-13.2%; p < 0.05) and long-term period (50.3+/-16.1%; p < 0.001); but the reference vessel diameter did not change significantly. Late restenosis occurred in 28% (3-5 years) and 33% (7-12 years) of 94 lesions, and 13.8% of lesion required repeat target lesion revascularization. During this period, 5.3% of patients (5 lesions) underwent revascularization for new proximal or distal lesions. CONCLUSIONS: Decrease of luminal diameter during the early 6 months, was followed by regression after stabilization of the balloon-dilated site up to 5 years, but luminal re-narrowing occurs again over 7 years after balloon angioplasty. PMID- 15791030 TI - C-reactive protein concentration is more strongly related to metabolic syndrome in women than in men: the Minoh Study. AB - BACKGROUND: The gender differences in the association between C-reactive protein (CRP) and features of the metabolic syndrome (MS) need to be elucidated among Japanese. METHODS AND RESULTS: The study population included 715 men and 988 women aged 40-69 years who were not taking anti-hypertensive, lipid-lowering, hypoglycemic, anti-thrombotic, or non-steroidal anti-inflammation medications, and did not have a past history of cardiovascular disease or CRP concentration >10 mg/L. Except for high-density lipoprotein cholesterol, the unadjusted correlation between CRP and each MS component, including body mass index (BMI), systolic and diastolic blood pressures, triglycerides, fasting glucose, fasting insulin, and uric acid, was greater in women than in men. With adjustment for age, smoking status, and drinking status, the differences in CRP concentrations between those with the MS components of BMI, triglycerides, and uric acid and those without were greater in women than in men. Results of stratified analyses by the number of components of the MS of 0, 1, 2, 3, and > or = 4 revealed that an increase in CRP concentrations was greater in women than men with an increased number of components of the MS (gender interaction, p = 0.005). This tendency was observed in non-smokers, but not in current smokers (gender interaction, p = 0.013 and = 0.513, respectively). CONCLUSIONS: CRP concentrations are closely related to the MS-like state in both sexes, but an increase in CRP concentration associated with risk factor-clustering is more pronounced in women, particularly non-smokers. PMID- 15791031 TI - Natural history of a dilated ascending aorta after aortic valve replacement. AB - BACKGROUND: Little information is available regarding the incidence of aortic dissection or rupture in patients with a dilated ascending aorta after aortic valve replacement (AVR). The present clinical study aimed to demonstrate the incidence of aortic complications after AVR in patients with a dilated ascending aorta and to clarify those risk factors associated with the progression of a dilated ascending aorta or late aortic events. METHODS AND RESULTS: A total of 35 patients with a dilated ascending aorta at the time of AVR were enrolled. A dilated ascending aorta was defined as 40 mm or greater in diameter by preoperative computed tomography or operative findings. The baseline ascending aorta diameter ranged from 40 to 55 mm with a mean of 44.8+/-4.4 mm. There was a high frequency of bicuspid valve disease in patients with a dilated ascending aorta (57%). The mean follow-up interval was 8.1+/-3.5 years (range: 2.3-13). Aortic events occurred in 5 patients (aortic dissection in 1, rupture in 2, reoperation in 2) during the follow-up. One aortic dissection developed at a baseline aortic size of 42 mm, whereas 2 aortic ruptures occurred at baseline aortic sizes of 47 mm and 50 mm. There was no statistically significant univariate association between any of the patient clinical characteristics and late aortic events or ascending aortic progression. CONCLUSION: Although the clinical course of patients with a dilated ascending aorta is unpredictable, aortic events may occur even in patients with a baseline aortic diameter of <50 mm. Therefore, preventive aortic surgery at the time of AVR should be considered to prevent aortic dissection or rupture in patients with an even slightly dilated ascending aorta with a diameter of 40 to 50 mm, unless the patient has a high operative risk or older age. PMID- 15791032 TI - A rapid bedside D-dimer assay (cardiac D-dimer) for screening of clinically suspected acute aortic dissection. AB - BACKGROUND: A rapid laboratory test for diagnosis of acute aortic dissection (AAD) has not been available. We performed this prospective study to determine the utility of a rapid bedside D-dimer (DD) assay for detection of AAD. METHODS AND RESULTS: Patients with suspected AAD were recruited and their DD levels were measured by rapid bedside assay. They were divided into 2 groups according to enhanced computed tomography findings: an AAD group (n = 30) and a non-AAD group (n = 48). The median DD level was higher in the AAD group (1.80 microg/ml) than in the non-AAD group (0.42 microg/ml) (p = 0.000). The rapid bedside DD assay showed 100% sensitivity, 54% specificity, 58% positive predictive value and 100% negative predictive value for detection of AAD with a normal DD level of up to 0.5 microg/ml. The combination of DD level >0.5 microg/ml and systolic blood pressure > or = 180 mmHg showed 86% positive predictive value for detection of AAD. Conclusions We conclude that the rapid bedside DD assay is a highly sensitive method for early exclusion of AAD in patients with chest and/or back pain suggestive of AAD. Acute aortic dissection is highly probable if a rapid DD assay shows the elevated DD level with systolic blood pressure > or = 180 mmHg on admission. PMID- 15791033 TI - Incidence of acute myocardial infarction in Takashima, Shiga, Japan. AB - BACKGROUND: The incidence and mortality from ischemic heart disease (IHD) in Japan seem to be among the lowest of all the industrialized countries, but there are few reliable registers of acute myocardial infarction (AMI). METHODS AND RESULTS: To assess the incidence of AMI in Takashima County, Shiga, Japan, from 1988 to 1998 and compare the data with similar registers in the world, cases of AMI or sudden death presumed from myocardial ischemia were registered. The criteria of AMI were based on the WHO MONICA Projects. The medical records of all the hospitals inside as well as outside the county, the original death records in the health center, and the ambulance records in the county were investigated and 291 cases were registered (190 males, 101 females; average age (mean +/- SD), 69.5+/-12.2). The 28-day and 24-h case fatality was 38.1% and 33.0%, respectively. Age-adjusted annual incidence of AMI per 100,000 population aged between 25 and 74 years were 58.2 for men and 18.0 for women. The incidence of AMI showed a constant trend from 1988 to 1998. CONCLUSION: The results confirmed that Japan has the lowest incidence of AMI among the industrialized countries. PMID- 15791034 TI - Sex difference in the relationship between insulin resistance and corrected QT interval in non-diabetic subjects. AB - BACKGROUND: Men with a prolonged corrected QT (QTc) interval have an increased risk of cardiovascular mortality and sudden death, even in healthy individuals. In addition, prolonged QTc is a predictor of mortality in diabetics. However, the relationship between insulin resistance and QTc is not clarified in non-diabetic healthy people. The present study was performed to observe the association between QTc and insulin resistance in Korean non-diabetic subjects. METHODS AND RESULTS: In the current study there was a total of 874 subjects (520 men, 354 women, mean age: 45.9+/-11.0 years) who underwent a medical check-up at the health promotion center at Kangbuk Samsung Hospital from January 2002 to May 2002. Age, sex, height, body weight, blood pressure, blood cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C), fasting insulin, and fasting glucose levels were measured. The QT intervals were corrected using Bazett's formula (QTc = QT/square root of RR). Homeostasis model assessments (HOMA) were performed to assess the correlation between insulin resistance indices and the QTc interval. The mean QTc interval was significantly longer in females (417+/-24 ms) than in males (402+/-23 ms) (p < 0.001). After adjusting the variables related to the QTc interval, the differences in QTc between men and women were statistically significant (p < 0.001). A significant positive correlation was found between QTc and age, glucose, and blood pressure in male subjects. Female subjects showed positive correlation between QTc and age, glucose, blood pressure, total cholesterol, low-density lipoprotein cholesterol, triglyceride, fasting insulin, and the HOMA index, and a negative correlation between QTc and HDL-C. Multiple regression analysis showed that in men, age (beta = 0.480, p < 0.001) and diastolic blood pressure (beta = 0.280, p < 0.001) were predictors of QTc. In women, age (beta = 0.321, p = 0.008), diastolic blood pressure (beta = 0.324, p = 0.006) and HOMA index (beta = 3.508, p = 0.033) were predictors of QTc. CONCLUSIONS: The present study of Korean healthy subjects shows that QTc was more prolonged in females than in males. In normoglycemic female subjects, insulin resistance was an independent determinant of the prolongation of QTc. PMID- 15791035 TI - Impact of preinterventional arterial remodeling on in-stent neointimal hyperplasia and in-stent restenosis after coronary stent implantation. AB - BACKGROUND: Patterns of arterial remodeling during the course of plaque development have been shown to play an important role in both the progression of de novo atherosclerosis and in the restenotic process following coronary intervention. The aim of the present prospective study was to evaluate the effect of pre-interventional arterial remodeling on in-stent neointimal hyperplasia (NIH) and in-stent restenosis (ISR) after stenting. METHODS AND RESULTS: Pre interventional arterial remodeling was assessed in 85 native coronary lesions by using intravascular ultrasound (IVUS). The remodeling index (RI) was 1.09+/-0.20 in the positive remodeling (PR)/intermediate remodeling (IR) group and 0.84+/ 0.12 in the negative remodeling (NR) group. The plaque plus media cross sectional area (P&M CSA) at pre-intervention and NIH CSA at follow-up in the minimal lumen CSA were significantly larger in the PR/IR group (9.2+/-2.9 mm2 vs 6.2+/-1.8 mm2, 3.3+/-1.2 mm2 vs 1.5+/-0.9 mm2; p = 0.001, p = 0.001, respectively). On 3 dimensional analysis of IVUS images at follow-up, the lumen volume was significantly smaller in the PR/IR group than that in the NR group (62+/-15 mm3 vs 75 +/-20 mm3; p = 0.001), and neointima hyperplasia volume was significantly larger in the PR/IR group than that in the NR group (46+/-15 mm3 vs 26+/-10 mm3; p = 0.001). A significant positive correlation was found between pre interventional RI and follow-up NIH CSA (r = 0.25, p = 0.022). The incidence of ISR and repeat intervention was significantly higher in the PR/IR group (30.8% vs 18.2%, 28.8% vs 15.2%; p = 0.032, 0.035, respectively). CONCLUSION: Measuring pre interventional arterial remodeling patterns by IVUS may be helpful to stratify lesions at high-risk of ISR. PMID- 15791036 TI - Recent decline in hospital mortality among patients with acute myocardial infarction. AB - BACKGROUND: Many patients with acute myocardial infarction will still die after admission. Recent trends in hospital mortality were analyzed to identify aspects that need improvement. METHODS AND RESULTS: A total of 1,247 patients admitted to Kinki University School of Medicine within 24 h of the onset of infarction were analyzed between 1975 and 2001. The percentage of patients discharged with 100% occlusion decreased gradually from 31.3% during 1975-1982 to 2.1% during 1998 2001, while those with 50% stenosis or less gradually increased from 12.5% to 82.5% during the same period (trends: p < 0.01). The cardiac death rate was 17.1% in 1975-1982, and 7.7% in 1998-2001, showing a significant decrease with time (p < 0.01). This decrease was particularly marked among those admitted within 6 h of the onset of infarction. Death due to cardiac rupture decreased significantly with time (p < 0.001). In contrast, the non-cardiac death rate, amounting to 2.2% on average, did not decline. CONCLUSIONS: Cardiac deaths due to acute myocardial infarction have decreased markedly of late. However, patients must be admitted within 6 h of the onset of infarction to benefit from this improvement. More effort should be made to improve the general care of patients in order to reduce the incidence of non-cardiac death. PMID- 15791037 TI - Spontaneous anterograde flow of the infarct artery preserves myocardial perfusion and fatty acid metabolism in patients with anterior acute myocardial infarction. AB - BACKGROUND: It remains unclear whether spontaneous anterograde flow preserves myocardial fatty acid metabolism in patients with acute myocardial infarction (AMI). METHODS AND RESULTS: The present study comprised 129 patients with a first anterior AMI in whom Thrombolysis in Myocardial Infarction (TIMI) 3 flow was obtained on the final angiogram: 28 patients with spontaneous anterograde flow and 101 patients with total occlusion on the initial angiogram. Thallium-201 (201Tl) and iodine-123-beta-methyl-p-iodophenyl penta-decanoic acid (123I-BMIPP) dual-isotope myocardial single-photon emission computed tomography (SPECT) was performed at 6.5+/-4.2 days after onset. The SPECT image was divided into 17 segments, and each segment was graded with scores between 0 and 4 (0, normal uptake; 4, defective). The sum of each score was defined as the total defect score (TDS). TDS values for 201Tl (9.0 +/-7.4 vs 16.8+/-12.2, p < 0.01) and 123I BMIPP (19.3+/-11.6 vs 24.1 +/-10.4, p < 0.05) were significantly lower in patients with spontaneous anterograde flow than in those with total occlusion. CONCLUSIONS: These results suggest that spontaneous anterograde flow of the infarct artery preserves not only myocardial perfusion but also fatty acid metabolism in patients with AMI. 123I-BMIPP SPECT image may underestimate the area at risk especially in patients with spontaneous anterograde flow. PMID- 15791038 TI - Can transthoracic Doppler echocardiography predict the discrepancy between left ventricular end-diastolic pressure and mean pulmonary capillary wedge pressure in patients with heart failure? AB - BACKGROUND: Left ventricular end-diastolic pressure (LVEDP) is difficult to measure continuously; therefore, pulmonary capillary wedge pressure (PCWP) is frequently used instead for hemodynamic monitoring in patients with heart failure. However, a discrepancy between LVEDP and mean PCWP is sometimes observed. METHODS AND RESULTS: To assess the feasibility of evaluating this discrepancy using echo-Doppler indexes, 140 consecutive patients with heart disease were studied. Transthoracic Doppler echocardiography (TTDE) was performed immediately before bilateral-sided cardiac catheterization. We measured peak velocities of early (E: cm/s) and late (A: cm/s) diastolic transmitral flow, and duration of A wave (MAd: ms). We also measured the duration of atrial reversal of pulmonary venous flow (PAd: ms). The difference between PAd and MAd (Deltad = PAd MAd: ms) was calculated. The ratio of E to tissue Doppler-derived peak early diastolic velocity of mitral annulus (Ea: cm/s) was also calculated (E/Ea). There was a good positive correlation between LVEDP and Deltad (r = 0.77, p < 0.001). There was a modest correlation between mean PCWP and E/Ea (r = 0.56, p < 0.001). When patients were classified by Deltad > or = 10 ms and E/Ea < or = 14, elevated LVEDP (> or = 17 mmHg) and normal mean PCWP (< or = 12 mmHg) were predicted with 100% sensitivity and 85% specificity. CONCLUSIONS: Evaluation of the discrepancy between LVEDP and mean PCWP in patients with heart failure was feasible by separately estimating LVEDP by Deltad and mean PCWP by E/Ea using noninvasive TTDE. Early detection of patients with elevated LVEDP and normal mean PCWP may be useful for preventing acute exacerbation of chronic heart failure. PMID- 15791039 TI - Effects of postischemic regional left ventricular diastolic wall motion abnormalities or delayed relaxation following coronary vasospasm on global diastolic function. AB - BACKGROUND: Regional left ventricular (LV) diastolic wall motion abnormalities detected by color kinesis (CK), an echocardiographic technique, may be a more sensitive measure to postischemic damage following coronary spasm than parameters of global diastolic function. METHODS AND RESULTS: Regional LV diastolic wall motion was evaluated by using CK in 18 patients with variant angina on the day following coronary spasm, which was induced by intracoronary acetylcholine. Fractional regional LV cavity area expansion in the short-axis view during the first 30% of the LV filling time, was used to identify postischemic asynchronous diastolic wall motion. Regional delayed relaxation was observed in any of the LV regions in all the patients, who were divided into 2 groups (Group S: 7 patients with single-vessel spasm with regional delayed relaxation in one area. Group M: 11 patients with multivessel spasm or spasm of the proximal left anterior descending branch with regional delayed relaxation in multiple areas). In Group S, no abnormality (0%) was noted in any of the indexes of global diastolic function including the isovolumic relaxation time, the ratio of peak rapid filling to peak atrial filling velocities and the deceleration time. In contrast, in 5 (45%) of the Group M patients, abnormalities were noted in all of those indexes. CONCLUSIONS: Postischemic regional LV-delayed relaxation following coronary vasospasm was detected sensitively by analysis of CK images. The indexes of global LV diastolic function are insensitive to postischemic damage following single vessel spasm, although they are somewhat sensitive following multivessel spasm. PMID- 15791040 TI - Left atrial appendage outflow velocity is superior to conventional criteria for predicting of maintenance of sinus rhythm after simple cryoablation of pulmonary vein orifices. AB - BACKGROUND: We sought to identify factors favoring long-term restoration of sinus rhythm (SR) in patients with atrial fibrillation (AF) who underwent a simple cryoablation of pulmonary vein orifices (PV-cryo) as part of their cardiac surgery. METHODS AND RESULTS: Of 101 patients with AF undergoing PV-cryo, the 71 in SR at discharge were grouped according to whether they maintained or lost SR (group SR, n = 61, and group AF, n = 10) after an average of 2.3 years. Atrial fibrillation present at discharge (n = 30) persisted during follow up. Comparisons were made to identify preoperative predictive factors, including transthoracic and transesophageal echocardiographic parameters. Of patients discharged from the hospital with SR, 92% (46 of 50) of those with AF duration of 3 years or less were in group SR, as were 92% (23 of 25) of those with left atrial dimension of 50 mm or less, and 93% (37 of 40) of those with average peak left atrial appendage outflow velocities (LAA-V) of at least 30 cm/s. Of 25 patients in group SR who had no paroxysmal AF and did not require antiarrhythmic drugs, all had LAA-V over 20 cm/s. Patients in group AF all had LAA-V under 40 cm/s. CONCLUSIONS: Left atrial appendage outflow velocities was the best predictor of whether SR was maintained long-term after PV-cryo. PMID- 15791041 TI - Association of inflammatory marker and highly sensitive C-reactive protein with aerobic exercise capacity, maximum oxygen uptake and insulin resistance in healthy middle-aged volunteers. AB - BACKGROUND: Increased levels of inflammation markers, such as C-reactive protein (CRP) and tumor necrosis factor-alpha , have been found in insulin resistance syndrome. Those with elevated levels of high-sensitive CRP (hs-CRP) are at a higher risk for coronary heart disease. In the present study, we evaluated whether maximum oxygen uptake and insulin resistance are related to hs-CRP for the primary prevention of coronary heart disease. METHODS AND RESULTS: The subjects were 50 subjects who did not have diabetes mellitus. A multi-step treadmill exercise test was performed to obtain the maximum oxygen uptake when assessed by computerized breath-by-breath analysis. As an index of insulin resistance, the homeostasis model insulin resistance index (HOMA-R; fasting glucose x fasting insulin/405) was used. In addition, bodyweight, body mass index, subcutaneous fat thickness, total cholesterol, high-density lipoprotein (HDL) cholesterol, and triglyceride were measured. Multivariate analysis revealed that hs-CRP was significantly correlated with HDL-cholesterol, uric acid, gamma glutamyl transpeptidase and maximum oxygen uptake. The maximum oxygen uptake showed the smallest odds ratio was in terms of the relationship with hs-CRP. CONCLUSIONS: The present study suggests that the development of exercising habits increases the maximum oxygen uptake. Furthermore, an elevated maximum oxygen uptake decreases HOMA-R and reduces the inflammatory marker CRP, suggesting that exercising habit plays an important role in the primary prevention of coronary heart disease. PMID- 15791042 TI - Clinical characteristics and treatment of constrictive pericarditis in Taiwan. AB - BACKGROUND: Constrictive pericarditis is an uncommon disease that prevents the normal diastolic filling of the heart and pericardiectomy is the only satisfactory treatment. METHODS AND RESULTS: The clinical characteristics and treatment of patients who underwent pericardiectomy for constrictive pericarditis (n = 23) were reviewed. Surgery was performed via left anterolateral thoracotomy plus transsternal extension in 3 patients, and median sternotomy in 20 patients. There were 2 deaths, resulting in an overall mortality rate of 8.7%. Of the 23 patients, 8 had Mycobacterium tuberculosis (Tb) infection, 2 had streptococcus infection, 1 had strongyloidiasis (Strongyloides stercoralis) and 1 developed the condition after a myocardial infarction; 2 patients underwent pericardial substitute insertion as post-heart surgery, and 3 patients had connective tissue disorders; 6 patients had idiopathic disease. CONCLUSION: These results show that bacterial infection, especially Tb, is a major etiology of constrictive pericarditis in Taiwan and that median sternotomy is an excellent approach for exposing the heart for pericardiectomy. PMID- 15791043 TI - Oral sildenafil improves primary pulmonary hypertension refractory to epoprostenol. AB - BACKGROUND: Epoprostenol (prostaglandin I(2)) has become recognized as a therapeutic breakthrough that can improve hemodynamics and survival in patients with primary pulmonary hypertension (PPH). However, a significant number of patients have PPH that is refractory to epoprostenol, and lung transplantation has been the only remaining treatment option. METHODS AND RESULTS: The study subjects included 20 consecutive patients with PPH (mean pulmonary arterial pressure: 65+/-15 mmHg) who had received epoprostenol for more than 12 months. The patients were divided into 2 groups; responders and non-responders. In the non-responders, New York Heart Association (NYHA) functional class did not improve and mean right atrial pressure (mRA) increased to 8 mmHg or more, and additional sildenafil, a phosphodiesterase-5 inhibitor, was started. Six patients were included in the non-responders, whose mRA was 9+/-5 mmHg before and significantly increased to 13+/-3 mmHg after epoprostenol administration (p < 0.05). One patient died and the other 5 patients received oral sildenafil. The mRA of 12+/-4 mmHg (value before sildenafil) improved to 8+/-5 mmHg after sildenafil administration. Three patients were classified in the NYHA functional class 4 and improved to class 3, and 2 patients were in class 3 and remained in the same class after the addition of sildenafil. CONCLUSIONS: In patients with severe PPH refractory to epoprostenol treatment, additional oral sildenafil can improve pulmonary hemodynamics and symptoms. The combination therapy of epoprostenol and sildenafil is a new medical treatment to attempt before progressing to lung transplantation for patients with PPH refractory to epoprostenol. PMID- 15791044 TI - Detection of amiodarone-induced pulmonary toxicity in supine and prone positions: high-resolution computed tomography study. AB - BACKGROUND: The aim of the present study was to describe the effectiveness and feasibility of high-resolution computed tomography (HRCT) in patients in supine and prone positions to detect amiodarone-induced pulmonary toxicity (APT). With regard to the possible differential diagnosis, our second goal was to emphasize the clinical value of HRCT with the patients in supine and prone positions compared with other paraclinical tests. METHODS AND RESULTS: Thoracic HRCT taken in both positions for 23 patients who were administrated amiodarone were prospectively evaluated in the current study. High-resolution computed tomography scans obtained with the patient in a prone position were helpful in differentiating dependent opacity from lung disease in 11 out of 23 patients. In another 4 patients, HRCT scans obtained with the patient in a prone position were useful in confirming the presence of subtle ground-glass opacities, considered as APT. Combination of HRCT in supine and prone positions provided a more reproducible method for evaluating the global extent of APT than other paraclinical tests. CONCLUSIONS: High-resolution computed tomography used in prone positions as well as a supine position could be an effective technique for reducing false-positive results in detection of APT and preventing the clinically serious pulmonary adverse effects by amiodanone. PMID- 15791045 TI - Is the chronotropic index applicable to children and adolescents? AB - BACKGROUND: The aim of the present study was to investigate whether the chronotropic index (CRI) is independent of age, sex, stage of exercise testing, functional capacity and resting heart rate in children, as it is in adult populations. METHODS AND RESULTS: A total of 129 normal children and adolescents underwent maximal treadmill exercise testing according to the Bruce protocol. Submaximal chronotropic response was assessed by the CRI at the end of stages 1, 2, 3, and 4 of Bruce treadmill exercise testing. The CRI assessed at each exercise stage was independent of sex, maximal oxygen uptake and resting heart rate. The CRI was independent of age when measured at stages 3 and 4 in boys and at stage 4 in girls, but was weakly correlated with age when measured at stages 1 and 2 in boys and at stages 1, 2 and 3 in girls. There was a significant effect of exercise intensity on the CRI: multiple comparisons revealed that, in girls, CRI assessed at stage 1 was greater than that assessed at stage 4. Also, lower exercise stages resulted in rather large variations in the measurement. CONCLUSION: The results essentially confirm the potential applicability of the CRI for children and adolescents, although care should be taken in evaluating measurements of the CRI in the early stages of exercise testing to avoid misinterpretation of the results. PMID- 15791046 TI - Fluvastatin ameliorates the hyperhomocysteinemia-induced endothelial dysfunction: the antioxidative properties of fluvastatin. AB - BACKGROUND: Hyperhomocysteinemia induces vascular endothelial dysfunction, contributing to a predisposition to the onset and/or progression of atherosclerosis. The major mechanism suggested for the adverse effect of homocysteine on vascular function seems to involve oxidative stress. Thus, we hypothesized that the administration of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor fluvastatin, which is experimentally demonstrated to have antioxidative properties as one of its pleiotropic effects, is a useful strategy for eliminating the detrimental events induced by hyperhomocysteinemia. METHODS AND RESULTS: In diet-induced hyperhomocysteinemic rats, we estimated oxidative stress and assessed endothelium-dependent vasodilatation. Hyperhomocysteinemia induced significant increases in urinary 8-isoprostaglandin F2alpha-III excretion and vascular superoxide generation, and impaired endothelium-dependent vasodilatation. Additional oral administration of the antioxidant fluvastatin or vitamin E, which normalized increased oxidative stress induced by hyperhomocysteinemia, ameliorated endothelial dysfunction. CONCLUSIONS: Hyperhomocysteinemia, even mild to moderate, induces endothelial dysfunction through its oxidative effect. The antioxidant fluvastatin was able to cancel out the oxidative stress induced by hyperhomocysteinemia and ameliorate endothelial dysfunction. Clinical use of fluvastatin might be a potent strategy for eliminating the detrimental events induced by hyperhomocysteinemia as well as hyperlipidemia. In addition to lowering homocysteine by means of folate supplementation, administration of the antioxidants is expected to be a potentially effective anti-homocysteine therapy. PMID- 15791047 TI - The mechanism of in-stent restenosis in radius stent: an experimental porcine study. AB - BACKGROUND: We investigated the mechanism of in-stent restenosis in radius stents in comparison to balloon-expandable stent (NIR stent) in pigs, with a focus on extracellular matrix (ECM). METHODS AND RESULTS: Radius (n = 4) or NIR (n = 4) stents were implanted in the left coronary arteries of miniature pigs. Quantitative coronary ultrasound (QCU) was performed before, immediately after, and at 1 and 4 weeks after the implantation. The stented-coronary arteries were harvested at 4 weeks after the implantation followed by immunohistochemical, histological, reverse transcription-polymerase chain reaction (RT-PCR) and real time PCR studies. In QCU, mean luminal areas at 4 weeks did not differ between both groups, whereas the mean stent area and neointimal area were significantly greater in the radius (p < 0.01). The immunohistochemical study revealed a significantly decreased number of neointimal macrophages and neovascularizations (p < 0.05, p < 0.01, respectively), and a stronger expression of tenascin-C in the radius. The histological study showed a larger ECM area and less neointimal cell density in the radius than in the NIR. The RT-PCR and real-time PCR analysis revealed an enhanced expression of tanascin-C mRNA in the radius than in the NIR. CONCLUSIONS: Increased production of ECM, especially tenascin-C, played a greater role in the neointimal formation in the radius stent than inflammation. PMID- 15791048 TI - Digital radiographic quantification of myocardial blood flow around a transmyocardial laser channel in rabbit hearts. AB - BACKGROUND: A mechanism underlying the benefits of transmyocardial laser revascularization (TMLR) has been presumed to be improvement in perfusion. We evaluated myocardial blood flow around a laser channel using digital radiography combined with a 3H-labeled desmethylimipramine ([3H]DMI) deposition. METHODS AND RESULTS: A laser channel was created in the left ventricular wall using a YAG laser in 6 non-ischemic rabbit hearts. After 8 weeks, [3H]DMI(1.11 MBq) was injected into the left atrium and the TMLR-treated myocardium was sectioned. Another 6 hearts were examined as controls. We measured [3H]DMI density in arbitrary units with digital radiography in the channel remnant, the surrounding area and a remote area. Flow distribution was quantified by the coefficient of variation of flows (CV). The surrounding area had the highest density (p < 0.001) and the lowest CV (p < 0.001), and had higher density (p < 0.001) and lower CV (p < 0.001) than the controls. There was no transmural difference in the density in all domains. The CV increased with depth in the remote area, as well as in controls (p < 0.001), but there was no transmural difference in the surrounding area. CONCLUSIONS: The TMLR increases myocardial blood flow and decreases flow heterogeneity in the surrounding area. The disappearance of transmural difference in flow heterogeneity might indicate the remodeling of microcirculation to improve regional oxygen delivery. PMID- 15791049 TI - Decreased vagal control over heart rate in rats with right-sided congestive heart failure: downregulation of neuronal nitric oxide synthase. AB - BACKGROUND: Parasympathetic drive is attenuated in heart failure, and resulting autonomic imbalance may increase the risk of sudden cardiac death. The anatomic site(s) and molecular mechanisms underlying this parasympathetic withdrawal are unknown. METHODS AND RESULTS: We examined the effects of pre- and post-ganglionic vagal nerve stimulation (VS) and acetylcholine (ACh) application on the heart rate of rats with right-sided congestive heart failure (CHF) induced by monocrotaline. Heart rate reduction in response to pre-ganglionic VS in CHF rats in vivo was significantly less than in controls. The suppression of spontaneous beating of isolated right atria including the whole sinoatrial (SA) node in response to post-ganglionic VS was significantly attenuated in CHF rats as well. In contrast, ACh application to the right atria resulted in a significantly larger suppression of spontaneous beating in CHF rats than controls. Proteins of neuronal nitric oxide synthase (nNOS) in the right atria were significantly decreased, whereas muscarinic (M2) receptor was significantly increased in CHF rats compared with controls. CONCLUSIONS: Both pre-and post-ganglionic vagal nerve functions are diminished in CHF rats, whereas M2 receptor-mediated regulation of the SA node is upregulated. Downregulation of nNOS may be involved in this parasympathetic withdrawal. PMID- 15791050 TI - Surgical case of isolated pulmonary Takayasu's arteritis. AB - Involvement of the pulmonary arteries is common in systemic Takayasu's arteritis, but pulmonary arteritis alone is very rare. A 67-year-old woman had severe right pulmonary artery stenosis and left pulmonary arterial obstruction caused by Takayasu's arteritis. Surgical treatment was successfully performed with extracorporeal circulation, without transection of the ascending aorta and the right pulmonary artery was enlarged with an autologous pericardial patch. PMID- 15791051 TI - Ruptured aneurysm of the subclavian artery in a patient with von Recklinghausen's disease. AB - A vascular manifestation of von Recklinghausen's disease is rare, but may be fatal if it is disrupted. A 39-year-old male with von Recklinghausen's disease presented with a large and tender bruise extending from his left neck to his left shoulder and arm. Computerized axial tomography revealed a ruptured aneurysm of the left subclavian artery. The patient underwent emergency surgery to exclude the aneurysm, and cross-over bypass grafting of both subclavian arteries was performed. We present a brief review of vascular maintestation of von Recklinghausen's disease, as well as a discussion regarding the strategy for treating this rare presentation. PMID- 15791052 TI - 'Gadolinium' as an alternative to iodinated contrast media for X-ray angiography in patients with severe allergy. AB - Some patients cannot undergo coronary angiography (CAG) because of the adverse effects of contrast media. In the present study gadolinium, (gadodiamide hydrate: Gd DTPA-BMA) commonly used for magnetic resonance imaging (MRI), was used as the contrast agent for diagnostic CAG in 3 patients with severe allergy to iodine or iodinated contrast agents. The indications for CAG were recurrent chest pain, evaluation of peri-operative risk of gastric cancer and abdominal aortic aneurysm, and evaluation of graft patency after coronary artery bypass grafting. The procedure was well tolerated by each patient and the amount of the gadolinium based contrast media did not exceed 40 ml, which is twice the volume used for MRI. The images were of fair quality. In patients with allergy to iodinated contrast media, CAG with gadolinium-based contrast media is an alternative technique for evaluating coronary artery disease. PMID- 15791053 TI - Permanent His-bundle pacing after atrioventricular node ablation in a patient with chronic atrial fibrillation and mitral regurgitation. AB - Hemodynamic deterioration because of worsening of mitral regurgitation can occur in a small number of patients undergoing atrioventricular node (AVN) ablation and pacing therapy. Patients with moderate mitral regurgitation before ablation seem prone to this complication. Successful permanent His-bundle pacing after AVN ablation was performed in a patient with chronic atrial fibrillation and moderate mitral regurgitation. Pulmonary capillary wedge pressure V-wave amplitude was markedly diminished and the mitral regurgitation area, calculated from the echocardiogram, was decreased by His-bundle pacing compared with that during right ventricular outflow tract or apical pacing. PMID- 15791056 TI - Examining assumptions about teen mothers: priorities and policies must set example for change. PMID- 15791055 TI - Surviving Social Security: it's 'America's Family Protection Plan.'. PMID- 15791057 TI - More on nurses and torture. PMID- 15791058 TI - Homosexual teens. PMID- 15791060 TI - More on tobacco cessation. PMID- 15791061 TI - More on tobacco cessation. PMID- 15791063 TI - Move over, Joe Camel: makers of alcoholic 'starter drinks' draw criticism. PMID- 15791065 TI - Treating asthma in pregnancy: what's good for women is good for fetuses, too. PMID- 15791067 TI - Emergency contraception for teens and young women: researchers find that it doesn't inhibit contraceptive use or increase rates of sexually transmitted infections. PMID- 15791068 TI - Sexually transmitted disasters? PMID- 15791069 TI - Nurse visits to moms and babies--a very good thing: program for at-risk mothers has long-term benefits. PMID- 15791071 TI - Improving patient care at the bedside: two new initiatives to create safer environments. PMID- 15791073 TI - Mercy for Banda Aceh: U.S. ship to provide full hospital services to tsunami stricken area. PMID- 15791074 TI - Proposed bills would transform nursing workplace and improve care. PMID- 15791075 TI - Reflections: dance with me?: loss and recovery on the farm. PMID- 15791076 TI - Cardiovascular disease in chronic kidney disease: recognizing and reducing the risk of a common CKD comorbidity. PMID- 15791078 TI - The state of the science: focus on IVs and HIV. PMID- 15791080 TI - Death by catheterization?: sudden, unexpected deaths of older adults are often not questioned. PMID- 15791081 TI - Pain control: ketamine: low doses may provide relief for some painful conditions. PMID- 15791082 TI - Diabetes under control: identifying and treating gestational diabetes mellitus: advances in screening and current interventions. PMID- 15791088 TI - A look back: nursing care of typhoid fever: the pivotal role of nurses at the Children's Hospital of Philadelphia between 1895 and 1910: how the past informs the present. PMID- 15791089 TI - When consent isn't informed enough: what's the nurse's role when a patient has given consent but doesn't fully understand the risks? PMID- 15791094 TI - Rebuilding the bridge: health and the environment are major nursing concerns. PMID- 15791091 TI - Testing the fate of APRN practice?: the ANA and other nursing groups meet to discuss a controversial Texas proposal. PMID- 15791096 TI - Anesthesia, amnesia, and the amygdala: reducing the fear of intraoperative awareness. PMID- 15791097 TI - Brain cell damage and S-100B increase after acute lung injury. PMID- 15791098 TI - Effects of skin pigmentation on pulse oximeter accuracy at low saturation. AB - BACKGROUND: It is uncertain whether skin pigmentation affects pulse oximeter accuracy at low HbO2 saturation. METHODS: The accuracy of finger pulse oximeters during stable, plateau levels of arterial oxygen saturation (Sao2) between 60 and 100% were evaluated in 11 subjects with darkly pigmented skin and in 10 with light skin pigmentation. Oximeters tested were the Nellcor N-595 with the OxiMax A probe (Nellcor Inc., Pleasanton, CA), the Novametrix 513 (Novametrix Inc., Wallingford, CT), and the Nonin Onyx (Nonin Inc., Plymouth, MN). Semisupine subjects breathed air-nitrogen-carbon dioxide mixtures through a mouthpiece. A computer used end-tidal oxygen and carbon dioxide concentrations determined by mass spectrometry to estimate breath-by-breath Sao2, from which an operator adjusted inspired gas to rapidly achieve 2- to 3-min stable plateaus of desaturation. Comparisons of oxygen saturation measured by pulse oximetry (Spo2) with Sao2 (by Radiometer OSM3) were used in a multivariate model to determine the interrelation between saturation, skin pigmentation, and oximeter bias (Spo2 - Sao2). RESULTS: At 60-70% Sao2, Spo2 (mean of three oximeters) overestimated Sao2 (bias +/- SD) by 3.56 +/- 2.45% (n = 29) in darkly pigmented subjects, compared with 0.37 +/- 3.20% (n = 58) in lightly pigmented subjects (P < 0.0001). The SD of bias was not greater with dark than light skin. The dark-light skin differences at 60-70% Sao2 were 2.35% (Nonin), 3.38% (Novametrix), and 4.30% (Nellcor). Skin pigment-related differences were significant with Nonin below 70% Sao2, with Novametrix below 90%, and with Nellcor at all ranges. Pigment-related bias increased approximately in proportion to desaturation. CONCLUSIONS: The three tested pulse oximeters overestimated arterial oxygen saturation during hypoxia in dark-skinned individuals. PMID- 15791099 TI - Sevoflurane versus halothane: postoperative maladaptive behavioral changes: a randomized, controlled trial. AB - BACKGROUND: The authors conducted a double-blind, randomized, controlled trial to determine whether the use of sevoflurane in children undergoing anesthesia and surgery results in a higher incidence of postoperative maladaptive behavioral changes as compared with halothane. METHODS: Children and their parents (n = 102) were randomly assigned to either a halothane group (n = 50) or a sevoflurane group (n = 52). The intraoperative anesthetic protocol was strictly controlled, and the postoperative analgesic consumption and pain levels were recorded. The effect of the group assignment on emergence status and maladaptive postoperative behavioral changes was assessed both by validated psychological measures and physiologic instruments (actigraphy) on postoperative days 1-7. Anxiety of the parent and child was also assessed, as was the child's postoperative recovery (Recovery Inventory). RESULTS: There were no group differences in preoperative state anxiety, postoperative analgesic requirements, postoperative pain, or the incidence of emergence delirium (P = not significant). Two-way repeated-measures analysis of variance showed no group differences in the incidence of postoperative maladaptive behaviors (F4,72 = 0.60, P = 0.701) or actigraphic variables such as percent sleep, number of night awakenings, and night awakenings that lasted for more than 5 min (P = not significant). CONCLUSION: The authors found no increased incidence of emergence delirium, maladaptive postoperative behavior changes, or sleep disturbances in children undergoing anesthesia with sevoflurane as compared with halothane. PMID- 15791100 TI - Tranexamic acid reduces intraoperative blood loss in pediatric patients undergoing scoliosis surgery. AB - BACKGROUND: Excessive bleeding often occurs during pediatric scoliosis surgery and is attributed to numerous factors, including accelerated fibrinolysis. The authors hypothesized that administration of tranexamic acid would reduce bleeding and transfusion requirements during scoliosis surgery. METHODS: Forty-four patients scheduled to undergo elective spinal fusion were randomly assigned to receive either 100 mg/kg tranexamic acid before incision followed by an infusion of 10 mg . kg . h during surgery (tranexamic acid group) or 0.9% saline (placebo group). General anesthesia was administered according to a standard protocol. Blood loss, transfusion requirements, coagulation parameters, and complications were assessed. RESULTS: In the tranexamic acid group, blood loss was reduced by 41% compared with placebo (1,230 +/- 535 vs. 2,085 +/- 1,188 ml; P < 0.01). The amount of blood transfused did not differ between groups (615 +/- 460 vs. 940 +/- 718 ml; P = 0.08). Administration of tranexamic acid was a multivariate predictor of blood loss, as was American Society of Anesthesiologists physical status and preoperative platelet count. No apparent adverse drug effects occurred in any patient. CONCLUSION: Intraoperative administration of tranexamic acid significantly reduces blood loss during spinal surgery in children with scoliosis. PMID- 15791101 TI - Tetanic stimulation of the peripheral nerve before transcranial electrical stimulation can enlarge amplitudes of myogenic motor evoked potentials during general anesthesia with neuromuscular blockade. AB - BACKGROUND: Neuromuscular blockade can suppress myogenic motor evoked potentials (MEPs). The authors hypothesized that tetanic stimulation (TS) of the peripheral nerve before transcranial stimulation may enhance myogenic MEPs during neuromuscular blockade. In the current study, the authors evaluated MEP augmentations by TS at different levels of duration, posttetanic interval, neuromuscular blockade, and stimulus intensity. METHODS: Thirty-two patients undergoing propofol-fentanyl-nitrous oxide anesthesia were examined. Train-of five stimulation was delivered to C3-C4, and MEPs were recorded from the abductor hallucis muscle. In study 1, TS with a duration of 1, 3, or 5 s was delivered at 50 Hz to the tibial nerve 1, 3, or 5 s (interval) before transcranial stimulation, and the effects of TS on MEP amplitude were evaluated. In study 2, TS-induced MEP augmentations were evaluated at the neuromuscular blockade level (%T1) of 50% or 5%. In study 3, MEP augmentations by TS at stimulus intensities of 0, 5, 25, and 50 mA were evaluated. RESULTS: The application of TS significantly enlarged the amplitudes of MEPs at the combinations of duration (3, 5 s) and interval (1, 3, 5 s) compared with those without TS. TS-induced MEP augmentations were similarly observed at %T1 of both 50% and 5%. TS-induced MEP augmentations were observed at stimulus intensities of 25 and 50 mA. CONCLUSIONS: The results indicate that TS of the peripheral nerve before transcranial stimulation can enlarge the amplitude of MEPs during general anesthesia with neuromuscular blockade. TS of the peripheral nerve can be intraoperatively applied as a method to augment myogenic MEP responses. PMID- 15791102 TI - Does preoperative coronary angioplasty improve perioperative cardiac outcome? AB - BACKGROUND: Percutaneous coronary intervention (PCI) is performed in patients with coronary artery disease who are undergoing major noncardiac procedures to reduce perioperative cardiac morbidity and mortality. However, the impact of this approach on postoperative outcome remains controversial. METHODS: The authors analyzed a cohort of 1,152 patients after abdominal aortic surgery in which 78 patients underwent PCI. A propensity score analysis was performed. Also, using a logistic regression model, the authors determined variables associated with a severe postoperative coronary event or a death in patients without PCI. Then, in patients with PCI, they compared the expected and observed outcome. RESULTS: Five variables (age > 75 yr, blood transfusion > 3 units, repeated surgery, preoperative hemodialysis, and previous cardiac failure) independently predicted (with 94% correctly classified) a severe postoperative coronary event, and five variables (age > 75 yr, repeated surgery, previously abnormal ST segment/T waves, previous hypertension, and previous cardiac failure) independently predicted (with 97% correctly classified) postoperative death. In the PCI group, the observed percentages of patients with a severe postoperative coronary event (9.0% [95% confidence interval, 4.4-17.4]) or death (5.1% [95% confidence interval, 2.0 12.5]) were not significantly different from the expected percentages (8.2 and 6.9%, respectively). When all patients were pooled together, the odds ratios of PCI were not significant. The propensity score analysis provided a similar conclusion. CONCLUSION: PCI did not seem to limit significantly cardiac risk or death after aortic surgery. PMID- 15791103 TI - Antioxidants reverse reduction of the human hypoxic ventilatory response by subanesthetic isoflurane. AB - BACKGROUND: In subanesthetic concentrations, volatile anesthetics reduce the acute hypoxic response (AHR), presumably by a direct action on the carotid bodies but by an unknown molecular mechanism. To examine a possible involvement of reactive oxygen species or changes in redox state in this inhibiting effect, the authors studied the effect of antioxidants on the isoflurane-induced reduction of the AHR in humans. METHODS: In 10 volunteers, the authors studied the effect of antioxidants (intravenous ascorbic acid and oral alpha-tocopherol) on the reduction by isoflurane (0.12% end-tidal concentration) of the AHR on a 3-min isocapnic hypoxic stimulus (hemoglobin oxygen saturation 86 +/- 4%). All subjects participated in three separate sessions in which the effects of the antioxidants (session 1), placebo (session 2), and sham isoflurane plus antioxidants (session 3) were tested on the (sham) isoflurane-induced effect on the AHR. RESULTS: Isoflurane reduced the acute hypoxic response from 0.82 +/- 0.41 l . min . % to 0.49 +/- 0.23 l . min . % and from 0.89 +/- 0.43 l . min . % to 0.48 +/- 0.28 l . min . % in sessions 1 and 2, respectively (mean +/- SD; P < 0.05 vs. control). This reduction of the AHR was completely reversed by antioxidants (AHR = 0.76 +/- 0.39 l . min . %; not significantly different from control, session 1) but not by placebo in session 2 (AHR = 0.50 +/- 0.30 l . min . %; P < 005 vs. control). Sham isoflurane or antioxidants per se had no effect on the hypoxic response. CONCLUSIONS: The data indicate that isoflurane may depress the AHR by influencing the redox state of oxygen-sensing elements in the carotid bodies. This finding may have clinical implications for patients who are prone to recurrent hypoxic episodes, e.g., due to upper airway obstruction, in the postoperative period when low-dose isoflurane may persist in the body for some time. PMID- 15791104 TI - Does the amygdala mediate anesthetic-induced amnesia? Basolateral amygdala lesions block sevoflurane-induced amnesia. AB - BACKGROUND: Amnesia for aversive events caused by benzodiazepines or propofol depends on the basolateral amygdala (BLA). Whether the amnesia of volatile anesthesia is also mediated through the BLA is unknown. If so, a general principle of anesthetic-induced amnesia may be emerging. Here, using an inhibitory avoidance paradigm, the authors determine whether BLA lesions prevent sevoflurane-induced amnesia. METHODS: Male Sprague-Dawley rats were separated into two groups: sham-operated controls (n = 22) and rats given bilateral N methyl-D-aspartate lesions of the BLA (n = 32). After a 1-week recovery, the rats were randomly assigned to be trained during either air or sevoflurane (0.3% inspired, 0.14 minimum alveolar concentration) exposure. Animals learned to remain in the starting safe compartment of a step-through inhibitory avoidance apparatus for 100 consecutive seconds by administering foot shock (0.3 mA) whenever they entered an adjacent shock compartment. Memory was assessed at 24 h. Longer latencies to enter the shock compartment at 24 h imply better memory. RESULTS: Sham-air (n = 10) animals had a robust memory, with a median retention latency of 507 s (interquartile range, 270-600 s). Sham-sevoflurane (n = 6) animals were amnesic, with a latency of 52 s (27-120 s) (P < 0.01, vs. sham-air). Both the air-exposed (n = 5) and the sevoflurane-exposed (n = 8) animals with BLA lesions showed robust memory, with latencies of 350 s (300-590 s) and 378 s (363 488 s), respectively. The latencies for both did not differ from the performance of the sham-air group and were significantly greater than the latency of the sham sevoflurane group (both P < 0.01). CONCLUSIONS: BLA lesions block sevoflurane induced amnesia. A role for the BLA in mediating anesthetic-induced amnesia may be a general principle of anesthetic action. PMID- 15791106 TI - Endotoxemia-induced lymphocyte apoptosis is augmented by a hyperinsulinemic euglycemic clamp. AB - BACKGROUND: Sepsis and endotoxemia are associated with lymphocyte apoptosis. This has been regarded as harmful, contributing to further immune suppression in already immune-compromised patients. Because normalization of blood glucose improves outcome in critically ill patients, the authors hypothesized that one of the effects of insulin and normoglycemia would be inhibition of lymphocyte apoptosis. Therefore, in this experimental study in pigs, the authors examined the separate and combined effects of acute endotoxemia and a hyperinsulinemic euglycemic clamp (HEC) on lymphocyte apoptosis. METHODS: After 60 min of stabilization, 38 anesthetized and mechanically ventilated pigs (weight, 35-40 kg) were divided (by randomization performed before the experiment) into four groups and were then studied for 570 min. Group 1 received no intervention. Group 2 received a HEC (5 mm p-glucose, insulin infusion rate of 0.6 mU . kg (-1). min( 1)) for 570 min. Group 3 received a lipopolysaccharide infusion for 180 min. Group 4 was given a combination of a HEC and a lipopolysaccharide infusion. After the 570-min study period, the pigs were killed, and tissue was sampled from the spleen and frozen. In four sections of each sample, the apoptosis of B and T lymphocytes were analyzed using stereologic methods: The number of apoptotic B and T cells was estimated by fluorescence immunohistochemistry with anti-active caspase-3 and either anti-CD21 (B lymphocytes) or anti-CD3epsilon (T lymphocytes). The number of apoptotic B and T lymphocytes was then compared using two-way analysis of variance, and the interaction between endotoxemia and the clamp (hyperinsulinemia and euglycemia) was investigated. RESULTS: Endotoxemia induced apoptosis of B (P < 0.001) and T lymphocytes (P = 0.016) in the spleen, and this effect was independent of the clamp. The ratios of apoptotic cells in the spleen tissue of pigs with and without endotoxemia were 2.4 (confidence interval, 1.7-3.4) and 1.6 (confidence interval, 1.1-2.2) for B and T lymphocytes, respectively. Independent of endotoxin infusion, HEC increased the number of apoptotic lymphocytes (P = 0.029 and P = 0.038 for B and T lymphocytes, respectively). The ratios of the number of apoptotic spleen cells in pigs treated and not treated with HEC were 1.5 (confidence interval, 1.0-2.1) and 1.5 (confidence interval, 1.0-2.1) for B and T lymphocytes, respectively. CONCLUSION: In this porcine model, both endotoxemia and a HEC increased the number of apoptotic B and T lymphocytes in the spleen. Contrary to our hypothesis, lymphocyte apoptosis during acute endotoxemia was augmented by a HEC. PMID- 15791105 TI - S-100 protein and neurohistopathologic changes in a porcine model of acute lung injury. AB - BACKGROUND: Survivors of acute respiratory distress syndrome exhibit neuropsychological sequelae that might be attributable to hippocampal damage. The authors sought to determine the effects of hypoxemia in a pig model of acute lung injury on the hippocampal region and the release of S-100 protein in comparison to a control group in which hypoxemia was induced by reducing the inspired oxygen fraction. METHODS: Hypoxemia was induced in 14 female pigs by repeated lung lavages (lung injury group; n = 7) or by reducing the inspired oxygen fraction (hypoxia-only group; n = 7). Hemodynamic variables, gas exchange, and serum concentrations of S-100 protein were measured at baseline, after induction of acute lung injury, and subsequently for 12 h. Animals were euthanized, and the brains were removed for histopathologic examination. RESULTS: Comparable blood gases were seen in both groups. Serum S-100 protein concentrations were comparable for both groups at baseline. At all other time points, S-100 concentrations were significantly higher in the lung injury group. Neuropathologic examination showed basophilic and shrunken neurons of the pyramidal cell layer in the hippocampal CA1 subregion of all pigs in the lung injury group. Few abnormalities were seen in the hypoxia-only group. CONCLUSIONS: The same degree of hypoxemia induced in a lavage model of acute lung injury results in greater brain damage assessed by S-100 protein and histopathologic findings when compared to a group in which hypoxemia at the same degree was induced by reducing the inspired oxygen fraction. This suggests that acute lung injury leads to neuropathologic changes independent of hypoxemia. PMID- 15791107 TI - Transcutaneous fluorescence dilution cardiac output and circulating blood volume during hemorrhagic hypovolemia. AB - BACKGROUND: Cardiac output and circulating blood volume are important parameters for assessing cardiac function in the intensive care setting and during major surgeries. The authors tested in an animal model of hemorrhagic hypovolemia the feasibility of measuring these parameters simultaneously by transcutaneous fluorescence monitoring of an intravenous bolus injection of indocyanine green. METHODS: Fluorescence dilution cardiac output was measured in seven anesthetized rabbits and compared to thermodilution cardiac output. The optical probe used to excite the indocyanine green fluorescence was in contact with the skin above the ear artery. Local heating enhanced blood perfusion of the measurement site. Cardiac output was measured during baseline conditions, during hemorrhagic hypovolemia, and after partial restoration of the blood volume with reinfused blood. Estimates of the circulating blood volume were simultaneously obtained from the analysis of the fluorescence dilution traces. RESULTS: Cardiac output measured by fluorescence dilution (thermodilution) averaged 455 +/- 16 (450 +/- 13) ml/min in baseline conditions and 323 +/- 15 (330 +/- 13) ml/min during hypovolemia. Fluorescence dilution cardiac output was linearly related to thermodilution cardiac output (slope = 1.13 +/- 0.05, ordinate = -50 +/- 19 ml/min, R = 0.92). Interanimal differences explained most of the variance between cardiac output estimates obtained with the two techniques. Circulating blood volume decreased from 204 +/- 5 ml in baseline conditions to 174 +/- 8 ml after bleeding and reflected blood volume changes in this acute bleeding-reinfusion model. CONCLUSIONS: The study extends the applicability of the fluorescence dilution technique for cardiac output measurement to hypovolemic conditions and demonstrates its ability to produce accurate estimates of the circulating blood volume in experimental animals. PMID- 15791108 TI - Classic benzodiazepines modulate the open-close equilibrium in alpha1beta2gamma2L gamma-aminobutyric acid type A receptors. AB - BACKGROUND: Classic benzodiazepine agonists induce their clinical effects by binding to a site on gamma-aminobutyric acid type A (GABAA) receptors and enhancing receptor activity. There are conflicting data regarding whether the benzodiazepine site is allosterically coupled to gamma-aminobutyric acid binding versus the channel open-close (gating) equilibrium. The authors tested the hypothesis that benzodiazepine site ligands modulate alpha1beta2gamma2L GABAA receptor gating both in the absence of orthosteric agonists and when the orthosteric sites are occupied. METHODS: GABAA receptors were recombinantly expressed in Xenopus oocytes and studied using two-microelectrode voltage clamp electrophysiology. To test gating effects in the absence of orthosteric agonist, the authors used spontaneously active GABAA receptors containing a leucine-to threonine mutation at residue 264 on the alpha1 subunit. To examine effects on gating when orthosteric sites were fully occupied, they activated wild-type receptors with high concentrations of a partial agonist, piperidine-4-sulfonic acid. RESULTS: In the absence of orthosteric agonists, the channel activity of alpha1L264Tbeta2gamma2L receptors was increased by diazepam and midazolam and reduced by the inverse benzodiazepine agonist FG7142. Flumazenil displayed very weak agonism and blocked midazolam from further activating mutant channels. In wild-type receptors activated with saturating concentrations of piperidine-4 sulfonic acid, midazolam increased maximal efficacy. CONCLUSIONS: Independent of orthosteric site occupancy, classic benzodiazepines modulate the gating equilibrium in alpha1beta2gamma2L GABAA receptors and are therefore allosteric coagonists. A Monod-Wyman-Changeux coagonist gating model quantitatively predicts these effects, suggesting that benzodiazepines minimally alter orthosteric ligand binding. PMID- 15791109 TI - Differential effects of bupivacaine and ropivacaine enantiomers on intracellular Ca2+ regulation in murine skeletal muscle fibers. AB - BACKGROUND: Increased intracellular Ca concentrations are considered to be a major pathomechanism in local anesthetic myotoxicity. Racemic bupivacaine and S ropivacaine cause Ca release from the sarcoplasmic reticulum of skeletal muscle fibers and simultaneously inhibit Ca reuptake. Examining the optical isomers of both agents, the authors investigated stereoselective effects on muscular Ca regulation to get a closer insight in subcellular mechanisms of local anesthetic myotoxicity. METHODS: R- and S-enantiomers as well as racemic mixtures of both agents were tested in concentrations of 1, 5, 10, and 15 mm. Saponin-skinned muscle fibers from the extensor digitorum longus muscle of BALB/c mice were examined according to a standardized procedure. For the assessment of effects on Ca uptake and release from the sarcoplasmic reticulum, agents were added to the loading solution and the release solution, respectively, and force and Ca transients were monitored. RESULTS: The effects of S-enantiomers on both Ca release and reuptake were significantly more pronounced than those of racemic mixtures and R-enantiomers, respectively. In addition, the effects of racemates were markedly stronger than those of R-enantiomers. With regard to Ca release, the effects of bupivacaine isomers were more pronounced than the isomers of ropivacaine. CONCLUSIONS: These data show that stereoselectivity is involved in alterations of intracellular Ca regulation by bupivacaine and ropivacaine. S enantiomers seem to be more potent than R-enantiomers, with intermediate effects of racemic mixtures. In addition, lipophilicity also seems to determine the extent of Ca release by local anesthetics. PMID- 15791110 TI - Weak polar interactions confer albumin binding site selectivity for haloether anesthetics. AB - BACKGROUND: Enflurane and isoflurane are structural isomers with different anesthetic potencies and side effects. It is not clear whether these differences are produced by differing occupancy of common protein binding sites or by occupancy of different sites, but the very similar molecular properties make the latter possibility unlikely. In this study, the authors examined binding site selectivity of these anesthetics in human serum albumin (HSA). METHODS: Binding of isoflurane and enflurane with HSA was determined with isothermal titration calorimetry. Competition with known ligands (propofol) allowed localization of binding sites within the HSA molecule. Molecular properties of isoflurane and enflurane were calculated. RESULTS: Isoflurane binds HSA with higher affinity but smaller total enthalpy than enflurane. Enthalpogram analysis suggested that isoflurane bound a single site, whereas enflurane bound two. Competition experiments indicated that enflurane and isoflurane share one binding site, which also binds propofol. The additional enflurane site binds propofol but not isoflurane. Increased salt concentration decreased the affinity for isoflurane but not for enflurane. The dipole moment of isoflurane is higher than that of enflurane, and the isoflurane binding site is more polar. CONCLUSION: These data indicate two binding sites of different character for the haloether anesthetics on HSA. One site is more polar and prefers isoflurane, presumably because of its larger dipole. The second site prefers the less polar enflurane. Therefore, weak polar interactions confer considerable selectivity, and differences in drug action may arise from occupancy of different protein sites. PMID- 15791111 TI - Traumatic brain injury and hemorrhagic hypotension suppress neuroprotective gene expression in injured hippocampal neurons. AB - BACKGROUND: After traumatic brain injury, memory dysfunction is due in part to damage to the hippocampus. To study the molecular mechanisms of this selective vulnerability, the authors used laser capture microdissection of neurons stained with Fluoro-Jade to directly compare gene expression in injured (Fluoro-Jade positive) and adjacent uninjured (Fluoro-Jade-negative) rat hippocampal neurons after traumatic brain injury and traumatic brain injury plus hemorrhagic hypotension. METHODS: Twelve isoflurane-anesthetized Sprague-Dawley rats underwent moderate (2.0 atm) fluid percussion traumatic brain injury followed by either normotension or hemorrhagic hypotension. Animals were killed 24 h after injury. Frozen brain sections were double stained with 1% cresyl violet and 0.001% Fluoro-Jade. RNA from 10 Fluoro-Jade-positive neurons and 10 Fluoro-Jade negative neurons, obtained from the hippocampal CA1, CA3, and dentate gyrus subfields using laser capture microdissection, was linearly amplified and analyzed by quantitative ribonuclease protection assay for nine neuroprotective and apoptosis-related genes. RESULTS: In injured CA3 neurons, expression of the neuroprotective genes glutathione peroxidase 1, heme oxygenase 1, and brain derived neurotrophic factor was significantly decreased compared with that of adjacent uninjured neurons. Superimposition of hemorrhagic hypotension was associated with down-regulation of neuroprotective genes in both injured and uninjured neurons of all subregions. Expression of apoptosis-related genes did not vary between injured and uninjured neurons, with or without superimposed hemorrhage. CONCLUSIONS: The authors show, in the first direct comparison of messenger RNA levels in injured and uninjured hippocampal neurons, that injured neurons express lower levels of neuroprotective genes than adjacent uninjured neurons. PMID- 15791112 TI - Randomized, double-blind study of the analgesic efficacy of morphine-6 glucuronide versus morphine sulfate for postoperative pain in major surgery. AB - BACKGROUND: Morphine-6-glucuronide (M6G) has promising preclinical characteristics and encouraging pharmacokinetic features for acute nociceptive pain. Early studies have produced a good safety profile when compared to morphine sulfate, although in surrogate pain models studies, a mixed picture emerged. A study to evaluate the efficacy and safety profile in a clinical setting was designed. METHODS: The authors conducted a double-blind, randomized, dose-finding study of patients scheduled to undergo major joint replacement. One hundred patients of both sexes were included, with 50 patients in each group. A loading dose of 10 mg of study medication was given intravenously at induction of anesthesia, and two further doses were allowed during surgery if required. Bolus doses via a patient-controlled analgesia system were given subcutaneously at 2 mg/dose and set at a 10-min lockout. Assessments of pain intensity and relief were recorded during the 24-h period. RESULTS: There were no statistically significant differences between the treatments for 24-h mean pain intensity. However, pain intensity was significantly higher in the M6G group than in the morphine group at 30 min and 1 h. There was no statistical difference in 24-h mean pain relief or retrospective pain scores at any time point during the 24-h period. The severity of sedation was significantly greater in the morphine group than in the M6G group at 30 min, 1 h, 2 h, and 24 h. Respiratory depression was greater in the morphine group than in the M6G group, and more patients in the morphine group withdrew from the study because of respiratory depression. CONCLUSIONS: Overall, M6G has an analgesic effect similar to that of morphine over the first 24 h postoperatively. However, M6G may be slower onset initially than morphine; therefore, a larger initial dose may be required. PMID- 15791113 TI - Efficacy and safety of single and repeated administration of 1 gram intravenous acetaminophen injection (paracetamol) for pain management after major orthopedic surgery. AB - BACKGROUND: Intravenous acetaminophen injection (paracetamol) is marketed in Europe for the management of acute pain. A repeated-dose, randomized, double blind, placebo-controlled, three-parallel group study was performed to evaluate the analgesic efficacy and safety of intravenous acetaminophen as compared with its prodrug (propacetamol) and placebo. Propacetamol has been available in many European countries for more than 20 yr. METHODS: After orthopedic surgery, patients reporting moderate to severe pain received either 1 g intravenous acetaminophen, 2 g propacetamol, or placebo at 6-h intervals over 24 h. Patients were allowed "rescue" intravenous patient-controlled analgesia morphine. Pain intensity, pain relief, and morphine use were measured at selected intervals. Safety was monitored through adverse event reporting, clinical examination, and laboratory testing. RESULTS: One hundred fifty-one patients (intravenous acetaminophen: 49; propacetamol: 50; placebo: 52) received at least one dose of study medication. The intravenous acetaminophen and propacetamol groups differed significantly from the placebo group regarding pain relief from 15 min to 6 h (P < 0.05) and median time to morphine rescue (intravenous acetaminophen: 3 h; propacetamol: 2.6 h; placebo: 0.8 h). Intravenous acetaminophen and propacetamol significantly reduced morphine consumption over the 24-h period: The total morphine doses received over 24 h were 38.3 +/- 35.1 mg for intravenous acetaminophen, 40.8 +/- 30.2 mg for propacetamol, and 57. 4 +/- 52.3 mg for placebo, corresponding to decreases of -33% (19 mg) and -29% (17 mg) for intravenous acetaminophen and propacetamol, respectively. Drug-related adverse events were reported in 8.2%, 50% (most of them local), and 17.3% of patients treated with intravenous acetaminophen, propacetamol, and placebo, respectively. CONCLUSION: Intravenous acetaminophen, 1 g, administered over a 24-h period in patients with moderate to severe pain after orthopedic surgery provided rapid and effective analgesia and was well tolerated. PMID- 15791114 TI - Spinal glucocorticoid receptors contribute to the development of morphine tolerance in rats. AB - BACKGROUND: Opioid analgesic tolerance is a pharmacologic phenomenon involving the mechanisms of cellular adaptation. Central glucocorticoid receptors (GRs) have been implicated in the cellular mechanism of neuronal plasticity that has many cellular steps in common with the mechanism of opioid tolerance. In a rat model of morphine tolerance, the authors examined the hypothesis that spinal GRs would play a significant role in the development of tolerance to the antinociceptive effect of morphine. METHODS: In experiment 1, each group of rats received the GR antagonist RU38486 (0.5 or 1 microg), the mineralocorticoid receptor antagonist spironolactone (3 microg), or a vehicle, given intrathecally with morphine (10 microg) twice daily for 6 days. In experiment 2, four groups of rats were used, and each group received intrathecally 10 microg morphine plus 5 micromol GR antisense oligodeoxynucleotide, sense oligodeoxynucleotide, mixed base oligodeoxynucleotide, or vehicle. Western blotting was used to examine the expression of GRs within the spinal cord dorsal horn. In experiment 3, the GR agonist dexamethasone (4 microg) was given intrathecally twice daily in combination with 10 microg morphine. For all experiments, the development of morphine antinociceptive tolerance was assessed using the tail-flick test. RESULTS: The development of tolerance to the antinociceptive effect of morphine was substantially attenuated when the GR antagonist RU38486 (1 > 0.5 microg > vehicle) but not spironolactone was coadministered with morphine for 6 days. A single treatment with RU38486 did not affect morphine antinociception, nor did it reverse morphine tolerance on day 7. A similar reduction of morphine tolerance was observed in those rats treated with a GR antisense oligodeoxynucleotide but not a sense or mixed-base oligodeoxynucleotide. The administration of the GR antisense oligodeoxynucleotide also prevented GR up-regulation within the spinal cord dorsal horn. Moreover, the GR agonist dexamethasone facilitated the development of morphine tolerance. CONCLUSIONS: The results indicate an important role of spinal GRs in the cellular mechanisms of morphine tolerance in rats and may have significant implications in clinical opioid therapy. PMID- 15791115 TI - Pulmonary atelectasis: a pathogenic perioperative entity. AB - Atelectasis occurs in the dependent parts of the lungs of most patients who are anesthetized. Development of atelectasis is associated with decreased lung compliance, impairment of oxygenation, increased pulmonary vascular resistance, and development of lung injury. The adverse effects of atelectasis persist into the postoperative period and can impact patient recovery. This review article focuses on the causes, nature, and diagnosis of atelectasis. The authors discuss the effects and implications of atelectasis in the perioperative period and illustrate how preventive measures may impact outcome. In addition, they examine the impact of atelectasis and its prevention in acute lung injury. PMID- 15791116 TI - The Apgar score has survived the test of time. AB - In 1953, Virginia Apgar, M.D. published her proposal for a new method of evaluation of the newborn infant. The avowed purpose of this paper was to establish a simple and clear classification of newborn infants which can be used to compare the results of obstetric practices, types of maternal pain relief and the results of resuscitation. Having considered several objective signs pertaining to the condition of the infant at birth she selected five that could be evaluated and taught to the delivery room personnel without difficulty. These signs were heart rate, respiratory effort, reflex irritability, muscle tone and color. Sixty seconds after the complete birth of the baby a rating of zero, one or two was given to each sign, depending on whether it was absent or present. Virginia Apgar reviewed anesthesia records of 1025 infants born alive at Columbia Presbyterian Medical Center during the period of this report. All had been rated by her method. Infants in poor condition scored 0-2, infants in fair condition scored 3-7, while scores 8-10 were achieved by infants in good condition. The most favorable score 1 min after birth was obtained by infants delivered vaginally with the occiput the presenting part (average 8.4). Newborns delivered by version and breech extraction had the lowest score (average 6.3). Infants delivered by cesarean section were more vigorous (average score 8.0) when spinal was the method of anesthesia versus an average score of 5.0 when general anesthesia was used. Correlating the 60 s score with neonatal mortality, Virginia found that mature infants receiving 0, 1 or 2 scores had a neonatal death rate of 14%; those scoring 3, 4, 5, 6 or 7 had a death rate of 1.1%; and those in the 8 10 score group had a death rate of 0.13%. She concluded that the prognosis of an infant is excellent if he receives one of the upper three scores, and poor if one of the lowest three scores. PMID- 15791117 TI - Postextubation laryngospasm in a patient with spasmodic dysphonia. PMID- 15791118 TI - GW280430A: pharmacodynamics and potential adverse effects. PMID- 15791119 TI - GW280430A. PMID- 15791123 TI - The right thing in the right place: lumbar plexus block in children. PMID- 15791126 TI - A safer and more effective intervention for radiculopathic pain. PMID- 15791124 TI - Of mice and men: should we extrapolate rodent experimental data to the care of human neonates? PMID- 15791127 TI - Avoid excessive sedation during cervical injections. PMID- 15791128 TI - Endotracheal tube damage during head and neck surgeries as a result of Harmonic Scalpel use. PMID- 15791130 TI - Interference between Extraneal peritoneal dialysis and the Accu-Chek blood glucose monitor. PMID- 15791131 TI - Nitroglycerin to facilitate insertion of a labor epidural. PMID- 15791132 TI - Transpharyngeal ultrasonography for cannulation of the internal jugular vein. PMID- 15791135 TI - Gene expression profiling: a new tool for pediatric neurology? PMID- 15791136 TI - The latest on infantile spasms. AB - PURPOSE OF REVIEW: This article reviews the most significant advances in the field of infantile spasm during the past year, with emphasis on best practise for treatment, and on some new etiological genetic and metabolic causes for the spasms, and new advances in the knowledge of tuberous sclerosis. RECENT FINDINGS: Up-to-date information comparing corticotrophin, oral steroids and vigabatrin shows that hormonal treatment is the most effective therapy in the short term. In a recent randomized trial, large doses of prednisolone were as effective as corticotrophin. There are insufficient data to recommend any treatment schedule for infantile spasms. Vigabatrin is the choice for infants with tuberous sclerosis. Visual field defects in (older) children seem to be as common as in adults. In animals, vigabatrin can induce apoptosis of the neurons in the developing brain. New rare factors associated with infantile spasms are mitochondrial diseases, mutations of the Aristales-related homeobax gene and posterior quadrantic dysplasia syndrome. The outcome in children with tuberous sclerosis and infantile spasms is better understood. SUMMARY: The accurate determination of etiology is now becoming increasingly possible. There is still a lack of consensus about the treatment of first choice for infantile spasms. However, recent data show that hormonal treatment is the most effective therapy in the short term. Frequency of visual field defects in children treated with vigabatrin should be studied in addition to the long-term outcome in general. Advances in our understanding of brain maturation, etiologies, mechanisms and genetics underlying catastrophic epilepsy may facilitate more effective pharmacologic interventions. PMID- 15791137 TI - Rett syndrome. AB - PURPOSE OF REVIEW: Nearly 70 reports on Rett syndrome were published in 2004. We have selected 51 articles, including clinical reports, on pathophysiology, genotype-phenotype correlation, and clinical and basic molecular biology studies. These articles explain how mutation of the gene (MECP2) for methyl-CpG-binding protein 2 causes the particular disorders of Rett syndrome, and also induces other neurodevelopmental disorders, clarifying the situation for future studies. RECENT FINDINGS: The role of X-chromosome inactivation has been clarified in animal experiments. New isoforms of MeCP2 have been discovered and its functional characteristics are under research. Understanding of the influence of the MECP2 mutation on other neurodevelopmental disorders has increased. However, there is no apparent progress in neurophysiological studies. SUMMARY: Clinical studies included the pathophysiology of stereotyped movement, and cardiac and respiratory disturbances, and there were four therapeutic trials including one for epilepsy. For genotype-phenotype correlation the role of X-chromosome inactivation was looked at and its basic mechanisms were studied extensively in animals. Characteristics of mutations in the C-terminus and the biological function of the new isoform, exon 1, were introduced. In studies on related neurodevelopmental disorders, a relationship is suggested between the MECP2 gene and autism-related gene, with overlapping pathways, but this is not common to other neurodevelopmental disorders. Developmental studies suggest an important role for MeCP2 in the formation and/or maintenance of synapses, and clarify the molecular biological aspects of Rett syndrome. However, early involvement of the aminergic neurons, suggested as the basic, pathognomonic lesion of Rett syndrome, has unfortunately not been investigated with the MECP2 mutation. PMID- 15791138 TI - Hereditary peripheral neuropathies of childhood. AB - PURPOSE OF REVIEW: The purpose of this review is to assist neurologists, paediatricians and other interested readers in following the expanding volume of information relating to the hereditary peripheral neuropathies of childhood. RECENT FINDINGS: During the last year, an exciting new potential therapy for hereditary sensory and motor neuropathy has been reported, and there has been a rapid expansion in our understanding of the genetic basis of a number of dominantly and recessively inherited neuropathies of childhood, most particularly in forms with pure motor or sensory and autonomic involvement. SUMMARY: Advances in our understanding of the molecular basis of the inherited neuropathies of childhood may provide greater insight into the pathogenesis of these disorders, hopefully identifying new therapeutic strategies for these lifelong conditions. PMID- 15791139 TI - Therapeutic hypothermia for hypoxic-ischaemic encephalopathy in the newborn infant. AB - PURPOSE OF REVIEW: This review examines recent findings from experimental models and clinical trials of induced hypothermia as treatment after cerebral hypoxia ischaemia in term newborn infants. RECENT FINDINGS: Experimental hypothermia inhibits many steps in the biochemical cascade that produces severe brain injury after hypoxia-ischaemia. This is in contrast to pharmacological agents, which tend to target only one step in the process that leads to brain injury. In adult humans hypothermia initiated immediately after cardiac arrest has improved outcomes. Delayed cooling after brain trauma has also been effective in a subgroup of adult patients. Seventy-two hours of selective head cooling with mild systemic hypothermia (rectal temperature 34.5 degrees C) in term infants with hypoxic-ischaemic encephalopathy (HIE) reduced death or disability in the infants with less severe electroencephalographic changes at entry (no benefit in those with advanced electroencephalographic changes). Cooling had no apparent adverse effects. A smaller randomized clinical trial of 48 h whole body cooling (rectal T 33 degrees C) found a reduction in death and neurological impairment. SUMMARY: In term infants with HIE there is emerging evidence that both selective head cooling and whole body cooling are neuroprotective and safe. This is consistent with a wealth of experimental animal data and adult trials. Neuroprotection seems to be lost if cooling is started after 6 h. The challenge now is to complete ongoing trials. If meta-analysis confirms a therapeutic effect, then this may lead to selection criteria and treatment protocols for very early hypothermia in HIE at term. PMID- 15791140 TI - Effect of inflammation on central nervous system development and vulnerability. AB - PURPOSE OF REVIEW: Preterm infants are at high risk for neurological sequelae and cognitive dysfunction. These problems have been attributed to a high occurrence of central nervous system (CNS) lesions, but suboptimal brain development appears to be just as important. In this brief review we present the hypothesis that systemic infection/inflammation can severely interfere with normal CNS function and development. RECENT FINDINGS: We focus on the effects of lipopolysaccharide because it is often used to model the systemic inflammatory response induced by infections. The inflammatory signals are propagated across the intact or ruptured blood-brain barrier to the CNS by proinflammatory cytokines, prostaglandins, or lipopolysaccharide. Subsequently, microglia are triggered to release cytokines, oxygen free radicals and trophic factors, which will influence the CNS in various ways. Cognition, dendritic length and spine density, dopaminergic cells, neurogenesis and glial proliferation will be affected. Furthermore, CNS vulnerability and, in some instances, cerebral anomalies and white matter damage are produced. SUMMARY: Hypothetically, all of these effects on the CNS triggered by inflammation may have severe consequences for the individual's ability to cope with environmental exposures during childhood and adulthood. PMID- 15791141 TI - The hidden causes of surgery-resistant temporal lobe epilepsy: extratemporal or temporal plus? PMID- 15791142 TI - Diagnosis and treatment of mood disorders in persons with epilepsy. AB - PURPOSE OF REVIEW: Epilepsy is a common, disabling neurological disorder associated with increased rates of comorbid psychiatric disorders as compared with the general population. RECENT FINDINGS: Mood disorders, especially major depression, appear to be more prevalent in persons with epilepsy than in those with the other chronic disorders and the general population. Depression may have more influence on quality of life than do cognitive and seizure factors. Although psychological, social, and vocational disabilities contribute to mood dysfunction in epilepsy, functional neuroimaging studies have consistently shown correlation of presence of cerebral abnormalities with increased severity of symptoms of depression. Most persons with epilepsy are not routinely screened for depression, and depression is subsequently treated in only a minority of patients. Although serotonin receptor density is greatest in brain regions commonly associated with epilepsy, such as the mesial temporal and prefrontal areas, no controlled trials have investigated the efficacy of serotonin reuptake inhibitors in persons with epilepsy. Optimal methods to identify and treat depression in epilepsy require substantial further research. SUMMARY: Depression is a common comorbid condition with significant negative effects on health status in persons with epilepsy, but additional understanding of the disorder is needed to improve diagnosis and treatment. PMID- 15791143 TI - Teratogenicity of antiepileptic drugs: state of the art. AB - PURPOSE OF REVIEW: Information on the risks associated with antiepileptic drug (AED) use in pregnancy is necessary for a rational approach to the management of women of childbearing potential who have epilepsy. This reviews addresses recent reports on the risk for birth defects and impaired postnatal development attributable to prenatal exposure to AEDs. RECENT FINDINGS: Some studies have indicated greater risk for birth defects as well as for lower verbal IQ in association with exposure to valproic acid, as compared with some other AEDs. The risk appears to be greater at doses above 800-1000 mg/day; lower doses may not carry higher risks than those associated with other AEDs. This observation is a cause of concern but more data are needed from prospective studies to assess the possible impact of confounding factors in order to clarify whether there is a causal relationship. Large-scale pregnancy registries have been launched and are expected to provide more conclusive comparative data on the most frequently used AEDs in the near future. SUMMARY: Although further studies are needed, it appears reasonable to use valproic acid with caution in women with epilepsy who are planning to become pregnant, and to consider prescribing other equally effective and safer AEDs if they are available. The importance of seizure control during pregnancy must not be neglected, and any attempt to change treatment should be done before conception. Drug withdrawal during pregnancy should be avoided, and patients should be made aware that definitive evidence on the relative safety of AEDs is lacking. PMID- 15791144 TI - The multiple facets of gamma-aminobutyric acid dysfunction in epilepsy. AB - PURPOSE OF REVIEW: The polarity of action of gamma-aminobutyric acid (GABA) changes from inhibition to excitation in the developing brain and in epilepsies. This review deals with recent observations concerning the mechanisms and clinical implications of the shift in GABA's activity from inhibition to excitation. RECENT FINDINGS: GABAergic synapses provide most transmitter-gated inhibition and are the targets of numerous clinically active agents, notably antiepileptic drugs. In a wide range of brain structures and species, GABAergic synapses are excitatory during maturation because of a higher concentration of intracellular chloride. These findings suggest that activation of GABA synapses will excite foetal neurones while inhibiting those of the mother. In epilepsies, recurrent seizures also lead to an accumulation of chloride and an excitatory action of GABA. These observations have major implications for clinical practice and research. They suggest that use of benzodiazepines by pregnant mothers may lead to deleterious consequences when they are taken during the period when GABA is the main excitatory transmitter. Because neuronal activity alters important cell functions, including migration and morphogenesis, aberrant excessive excitation may lead to profound deleterious consequences. SUMMARY: In several physiological and pathological conditions, activation of GABAergic synapses excites neurones instead of producing classical inhibition. This shift, which is due to an intracellular accumulation of chloride, has major consequences for both the operation of networks and the pathogenic effects of epilepsies. This is particularly important in the immature brain, where the excitatory actions of GABA are particularly prominent. PMID- 15791145 TI - Mechanisms, genetics, and pathogenesis of juvenile myoclonic epilepsy. AB - PURPOSE OF REVIEW: This review addresses the mechanisms, genetics and pathogenesis of juvenile myoclonic epilepsy (JME, Janz syndrome). RECENT FINDINGS: Although JME is a well defined clinical syndrome among the idiopathic generalized epilepsies (IGEs), recent studies suggest that JME is distinct from other IGE syndromes and must be considered separately for the purposes of genetic studies. Clinical, morphological and metabolic data suggest a preferential role for frontal regions in this syndrome. However, JME is clinically and genetically heterogeneous. Although several major genes for JME have been identified and pathogenetic mechanisms suggested based on these findings, these genes account for only a small proportion of JME cases, suggesting multifactorial or complex inheritance in most. The roles played by other major genes, susceptibility genes and environmental factors in the pathogenesis of JME remain to be defined. SUMMARY: JME is clinically and genetically heterogeneous and should be considered separately from other IGE syndromes. Proposed mechanisms, such as those involving microdysgenesis or altered neuronal inhibition, may be related to different genetic abnormalities in different patients. Major genes account for relatively few cases, and most cases appear to involve multifactorial or complex inheritance. PMID- 15791146 TI - Photosensitivity and visually induced seizures. AB - PURPOSE OF REVIEW: Interest in visually induced seizures has increased in recent years as a result of the increasing number of precipitants in our modern environment. This review addresses new developments in this field with special attention given to the emergence of new diagnostic, therapeutic and preventive approaches; it also emphasizes the importance of this condition as a public health issue. RECENT FINDINGS: Current evidence indicates the presence of two different mechanisms of photosensitivity, one dependent on luminance changes and the other on wavelength. Both mechanisms may be active in the same patient, although one may be dominant. Magnetoencephalography studies revealed an enhancement in gamma frequency preceding the development of a paroxysmal response as well as underlying uncomfortable visual illusions, suggesting that a loss of control over high-frequency oscillatory processes may be involved in the genesis of both types of phenomenon. The genetics underlying this trait remain to be determined. More precise definition of different phenotypes should help in this search. Recognition of the risks posed by the audiovisual environment for induction of seizures in photosensitive individuals, who may not even be aware of their condition, will prompt further development of guidelines and devices designed to prevent the occurrence of seizures triggered by dangerous video sequences. SUMMARY: Photosensitive epilepsy constitutes a unique benchmark model in which to address important issues in human epileptogenesis. The scope of the health risks posed by the modern audiovisual environment is increasingly being recognized, and further development of guidelines and regulations to control exposure to provocative materials are warranted. PMID- 15791147 TI - Functional magnetic resonance imaging for assessment of language and memory in clinical practice. AB - PURPOSE OF REVIEW: There has been considerable interest in the role that functional magnetic resonance imaging (fMRI) may play in the assessment of patients with epilepsy. This review considers recent progress in this field and the current role of fMRI in the preoperative assessment of language and memory function. RECENT FINDINGS: Many studies have compared fMRI with the intracarotid amytal test for establishing language dominance, with most showing over 90% concordance between the two tests. Atypical dominance is greater in patients with left temporal lobe epilepsy and has been shown to be associated with increased epileptic activity. Preoperative fMRI has been used to predict language deficits following left anterior temporal lobe resection. A variety of paradigms have been used to assess memory function, and novel paradigms have demonstrated robust medial temporal lobe activation. Different patterns of encoding activity have been observed between patients with unilateral temporal lobe epilepsy and control individuals. SUMMARY: The application of fMRI paradigms used in cognitive neuroscience to patients with epilepsy is complicated by several factors. Although fMRI is increasingly being used clinically to establish language dominance, further work is required to localize accurately those specific language functions that are most at risk following surgery. Memory paradigms are not yet validated for use in surgical planning, although methodological and technical advances should make this possible in the near future. Further studies looking at the reorganization of language and memory function after surgery are also required. PMID- 15791148 TI - Sexual hormones and epilepsy: threat and opportunities. AB - PURPOSE OF REVIEW: This article reviews recent developments in our knowledge of the reciprocal interactions between epilepsy and sex hormones and how these interactions may play a role in the pathophysiology and treatment of both. RECENT FINDINGS: Community studies confirm that menstrual disorders are overrepresented among women with epilepsy, especially among women with high seizure frequency and in those on valproate or polytherapy. Reproductive function is not affected in women with epilepsy who discontinued antiepileptic drug therapy during pubertal maturation. While valproate has been implicated as having particularly notable disruptive effects on reproductive function in women with epilepsy (polycystic ovaries and hyperandrogenemia), this was not evident in non-epileptic primates. The role of epilepsy itself is evident from a study that showed that the laterality of unilateral temporolimbic discharges is associated with predictable directional changes in hormonal secretion at all levels of the reproductive neuroendocrine axis. Epilepsy in men is associated with reduced levels of sexual function, bioactive testosterone and sperm. Various antiepileptic drugs may differ in this regard. SUMMARY: Epilepsy and antiepileptic drugs can alter sex hormone levels to promote the development of reproductive endocrine disorders in both women and men. Reproductive endocrine disorders may adversely affect both reproductive function and seizure control. Treatment of epilepsy and selection of antiepileptic drugs may be important to reproductive health in women and men with epilepsy. Sex steroids and their metabolites may also provide treatment for seizures. PMID- 15791149 TI - Progressive hippocampal and extrahippocampal atrophy in drug resistant epilepsy. AB - PURPOSE OF REVIEW: This article reviews recent experimental and clinical evidence for seizure-related progressive brain damage and discusses possible mechanisms of ongoing brain atrophy in epilepsy. RECENT FINDINGS: Experimental data indicate that seizures induce brain plasticity that may result in either damage or protection. Brief seizures or status epilepticus may promote resistance to additional damage but also induce cumulative neuronal loss and increase susceptibility to network synchronization. Some experimental studies indicated that, following the initial damage caused by status epilepticus, further brief seizures may not produce significant continuing neuronal loss and hippocampal atrophy, whereas other studies showed the contrary. There is recent evidence that progressive damage and atrophy may occur after an acute insult but are not directly associated with recurrent seizures. Clinical research data continue to show discrepancies regarding whether ongoing seizures cause progressive atrophy. Some cross-sectional and longitudinal magnetic resonance imaging studies in patients with partial epilepsies have shown progressive hippocampal and extrahippocampal atrophy, the severity of which correlated with the duration of epilepsy, seizure frequency, or lifetime seizure number, whereas others have failed to show a clear association. SUMMARY: Experimental data indicate that epileptogenesis in developing brain may not require significant neuronal loss, which is in keeping with clinical observations that progressive cognitive and behavioural impairment may occur in patients with no detectable brain atrophy. A better understanding of why, when and how progressive brain atrophy occurs will lead to better clinical management, earlier surgical intervention when necessary and, ultimately, prevention of epileptogenesis. PMID- 15791150 TI - Cerebral small vessel diseases: cerebral microangiopathies. AB - PURPOSE OF REVIEW: Small vessel diseases of the brain are still clinically underrecognized whereas their burden is increasing steeply. The most frequent is acquired degenerative small vessel disease. Many hereditary or idiopathic small vessel diseases have also been identified, of which cerebral autosomal-dominant arteriopathy with stroke and ischaemic leukoencephalopathy (CADASIL) is the most prominent. In this review, we will highlight current evidence on pathophysiology and genetics, new imaging tools, and treatment options. RECENT FINDINGS: Recent imaging studies have stressed the disruption of white matter connections in the pathogenesis of cognitive impairment in acquired small vessel disease. Clinical trials suggest a therapeutic benefit of acetylcholinesterase inhibitors. CADASIL is caused by mutations in the Notch3 gene. Current basic research has identified Notch genes to be important for endothelial and smooth muscle cells to form arteries and veins. Diagnosis can now be made reliably by magnetic resonance brain imaging, skin biopsy, or genetic testing. The so-called retinocerebral vasculopathies share the involvement of both retinal, cerebral and cochlear arterioles (e.g. Susac's syndrome). Cerebral amyloid angiopathy, occurring either sporadically or as a cause of various gene mutations, predisposes to lobar haemorrhages caused by rupture of affected small cortical vessels. Other rare cerebral microangiopathies, such as mitochondrial cytopathies, Fabry's disease and toxemic vasculopathy, offer established or new therapeutic options. SUMMARY: Cerebral small vessel diseases are highly variable in their aetiopathogenesis and clinical course. Current pathophysiological insights will help develop better treatment modalities; new imaging tools will provide surrogate markers for monitoring disease progression and treatment effects. PMID- 15791151 TI - Bibliography. Current world literature. Developmental disorders. PMID- 15791153 TI - Preface. PMID- 15791154 TI - Epidemiology of uveitis. PMID- 15791155 TI - Genetics in uveitis. PMID- 15791156 TI - Imaging techniques in the diagnosis and management of uveitis. PMID- 15791158 TI - Ocular tuberculosis: anterior segment. PMID- 15791159 TI - Tubercular posterior uveitis. PMID- 15791160 TI - Herpetic posterior uveitis. PMID- 15791157 TI - Polymerase chain reaction in the diagnosis of uveitis. PMID- 15791161 TI - Retinal vasculitis. PMID- 15791162 TI - Vogt-Koyanagi-Harada disease. PMID- 15791163 TI - Uveitis in children. PMID- 15791164 TI - Management of choroidal neovascularization in uveitis. PMID- 15791165 TI - Anti-tumor necrosis factor alpha antibody in the treatment of Behcet's disease. PMID- 15791166 TI - The diagnosis and management of anterior scleritis. PMID- 15791168 TI - Concerns over antidepressant medications and suicide: what does it mean for ECT? PMID- 15791169 TI - Electroconvulsive therapy in Belgium: a questionnaire study on the practice of electroconvulsive therapy in Flanders and the Brussels Capital region. AB - OBJECTIVES: We sought to review and describe the practice of electroconvulsive therapy (ECT) in Flanders and the Brussels Capital region, Belgium, in 2003. METHODS: A 30-item questionnaire was sent to all institutions that provided psychiatric care. Besides questions on training and general attitude toward ECT, most questions concerned technical aspects, quality, and organization of this treatment modality. RESULTS: All psychiatric departments offering ECT and 58% of the hospitals that do not conduct ECT replied to the questionnaire. Forty-three percent of the departments offering ECT indicated less than 10 treatment sessions per month. Forty-three percent of hospitals set a fixed number of treatments per course. Maintenance ECT and ECT on an outpatient basis were rather rare, except for a few departments. The major indication for ECT was depression (88%) rather than schizophrenia (8%), mania (3%), or other psychiatric disorders (1%). Propofol was the anesthetic most commonly used (74%). Seven departments (30%) used a sine wave device. Bilateral electrode placement was the preferred option in 91% of all departments and both fixed high dose and dose titration strategy were used equally (both 47.8%). CONCLUSION: The present findings demonstrate that the practice of ECT in Flanders and the Brussels Capital region in 2003 generally was not performed according to the available guidelines. ECT is underused in Flanders and the Brussels Capital region compared with other countries, although it is available in both specialized psychiatric facilities and in the psychiatric departments of general hospitals. PMID- 15791170 TI - The apolipoprotein E polymorphism is not associated with response to electroconvulsive therapy in major depressive disorder. AB - The apolipoprotein E (APOE) polymorphism is associated with neurodegenerative diseases. Its role regarding psychiatric disorders is controversial. It has been suggested to affect antidepressant treatment response and response to electroconvulsive therapy (ECT). In the present study, the association between APOE polymorphism and response to ECT in 119 patients with major depressive disorder was investigated. Moreover, a relation between APOE polymorphism and the age of onset of depression as well as the cognitive outcome of ECT was studied. In the whole population, no association was found between APOE polymorphism and response to ECT. However, in nonpsychotic patients, the epsilon2 allele tended to be more frequent in responders than nonresponders. Earlier onset of depression was observed in the patients with epsilon4 allele in late-life depression. There was no association between the APOE genotype and the cognitive change caused by ECT in the population as a whole. In women, however, epsilon2 allele may play a protective and epsilon4 allele a deleterious role in cognition during ECT. PMID- 15791172 TI - Seizure length and clinical outcome in electroconvulsive therapy using methohexital or thiopental. AB - Seizure duration is an extensively studied and controversial indicator of treatment quality in electroconvulsive therapy. Previous research comparing the effect of the barbiturate anesthetics methohexital and thiopental on seizure duration has yielded conflicting results. A recent period of unavailability of methohexital in the United States allowed for retrospective comparison of seizure length as well as clinical improvement in treatment using each agent. Retrospective review was made of 837 treatments administered to 97 patients between January 2, 2002, and May 31, 2003, examining anesthetic, seizure duration, and Global Assessment of Functioning (GAF) scores of inpatients at hospital admission and discharge. Analysis of variance of treatments 2-5 showed no significant effect for anesthetic on seizure duration. Analysis on a treatment by-treatment basis revealed a marginally significant trend toward shorter EEG seizures in the thiopental group at the second treatment (50.5 +/- 23.6 s vs. 61.1 +/- 27.9 s; P = 0.07) and fifth treatment (41.7 +/- 16.9 s vs. 51.8 +/- 24.0 s; P = 0.07). A difference approaching statistical significance revealed shorter convulsion length in the thiopental group at treatment 5 (29.0 +/- 12.3 s vs. 34.8 +/- 12.3 s; P = 0.07). Comparison of GAF score improvement at hospital discharge revealed no significant difference (GAF increase 26.4 +/- 9.4 for methohexital-treated patients vs. 24.8 +/- 12.0 for thiopental-treated patients; t = 1.00, df = 82, P > 0.1). Trends approaching significance in treatments 2 and 5 revealed shorter seizures in the thiopental group. However, data on clinical recovery reveals no greater improvement in the methohexital group. Thus, this study calls further into question the premise that choice of barbiturate anesthetic may affect clinical efficacy. PMID- 15791171 TI - The comparative hemodynamic effects of methohexital and remifentanil in electroconvulsive therapy. AB - Remifentanil is a short acting opioid frequently used to supplement general anesthesia for brief procedures. Narcotic agents are known for their ability to blunt autonomic responses to stimuli such as laryngoscopy and intubation and do not alter seizure threshold. We hypothesized that the combination of remifentanil and methohexital for induction would produce favorable suppression of sympathetic response during electroconvulsive therapy (ECT). With Institutional Review Board approval and informed consent, patients were enrolled in a prospective, randomized, double-blind, crossover study of methohexital alone versus remifentanil with an adjuvant of low-dose methohexital. One hundred ten ECT treatments were evaluated and subjects were treated in an alternating fashion with one of two induction protocols: Methohexital alone in an 80-100 mg IV bolus or remifentanil 500 mcg IV bolus combined with methohexital 40 mg IV. Bilateral ECT was performed in standard fashion and systolic blood pressure and heart rate were recorded throughout the procedure. No significant differences were found in baseline hemodynamic values between the two groups. Heart rate was significantly lower in the remifentanil group versus methohexital group at one minute post induction and just prior to ECT stimulus. Pre-ECT systolic blood pressure was not significantly different between the two groups. Heart rate remained lower in the remifentanil group at all measured timepoints during the treatment and continuously for five minutes after the seizure. Systolic blood pressure was significantly lower at one minute following the end of seizure and five minutes after end of seizure. Remifentanil's short duration of action, favorable side effect profile, potential proconvulsant activity and ability to suppress hemodynamic response make it a potential novel drug for ECT induction. PMID- 15791174 TI - Transcranial magnetic stimulation effects on one-trial learning and response to anxiogenic stimuli in adult male rats. AB - Transcranial magnetic stimulation (TMS) is a relatively new technique for inducing small, localized, and reversible changes in living brain tissue and has been suggested to have antidepressant properties in humans and animal models of depression. Memory function generally has been found to be unaffected by TMS, although some studies have raised the possibility of memory interference from TMS. Additionally, there have been indirect indications that TMS may possess anxiolytic features. This study examines the effects of TMS in animal models of one-trial learning and anxiety. In this study, short-term treatment with TMS compared with identically handled animals not given TMS in adult rats resulted in no significant differences in memory as assessed both by a one-time learning paradigm and by components of an elevated-plus maze task, that TMS does not impair memory as assessed by these tasks. In addition, no changes were found in anxiety-like behavior on the elevated plus maze task. In summary, these findings support previous reports that TMS does not interfere with memory function. There was no evidence of an anxiolytic response from TMS in rats as assessed by the elevated plus maze test. PMID- 15791173 TI - Enhanced attention-dependent auditory processing by electroconvulsive therapy in psychotic depression. AB - There is a lack of knowledge about the effect of electroconvulsive therapy (ECT) on auditory event-related potentials (ERPs) in severe psychotic depression. The aim of this study was to investigate both the effect of ECT on attention dependent ERP (P300) and the correlation of P300 values with depression level. We recorded the auditory ERPs of 23 patients expressing psychotic symptoms and fulfilling the DSM-III-R criteria for treatment-resistant severe major depressive episode before and a week after successful bitemporal ECT. The clinical status was assessed with Hamilton (Ham-D) and Montgomery and Asberg (MADRS) depression scales. ECT was clinically very effective with these scales. On the level of auditory processing, ECT increased P300 amplitude with no significant effect on latency. Small amplitudes over the left hemisphere before treatment were associated with bigger Ham-D-score decrement. ECT produces a significant increase in brain activity at the level of attention-dependent auditory processing in severe depression. The change in electrical responses seems to represent a largely independent variable from the clinical assessment scales, even if the recovery rate was remarkable, because the change in overall symptom scores did not correlate with the P300 changes. PMID- 15791175 TI - Efficacy of electroconvulsive therapy in treatment-resistant bipolar disorder: a case series. AB - The response to electroconvulsive therapy for six bipolar patients after pharmacotherapy failure is discussed. METHODS: Inclusion criteria were as follows: (1) bipolar mood disorder, manic, depressive or mixed episode (DSM-IV); (2) failure of pharmacotherapy, that is, for mania, manic episode unresponsive to at least 2 adequate antimanic trials for 6 weeks; and for bipolar depression, bipolar depressive episode unresponsive to at least 2 adequate antidepressant trials for 8 weeks. The patients underwent 12 bilateral sessions of ECT 3 times per week. Clinical response was considered a reduction of 50% or greater in the Young Mania Rating Scale (YMRS) and in the Hamilton Rating Scale for Depression 21 items (HAMD-21). Final YMRS <6 and HAMD-21 <8 defined remission. RESULTS: Six of the 9 referred patients consented to be submitted to ECT. Four were male, with ages ranging from 29 to 61 years, and their age at onset ranged from 19 to 49 years. Four showed psychotic features. All responded to ECT. PMID- 15791176 TI - Pulmonary embolism after ECT. AB - Electroconvulsive therapy (ECT) is a safe procedure, infrequently associated with life-threatening complications. Pulmonary embolism (PE) as a complication of ECT has been rarely reported. We describe a nonfatal case of PE that developed during ECT in a 50-year-old man with depression, hypertension, and diabetes. He developed symptoms of PE immediately upon awakening from the eighth right unilateral ECT, which was later confirmed by spiral chest computed tomography. We review the literature, discussing risk factors relevant to the pathophysiology of PE and making suggestions about the management of patients with suspicious symptoms. PMID- 15791177 TI - Chronic renal failure, hemodialysis, and electroconvulsive therapy: a case report. AB - We present a case study of a 60-year-old man with history of chronic renal failure on hemodialysis with severe bipolar I depression. Referred for worsening psychotic depression associated with refusal of oral intake and severe hypokinesia unresponsive to several antidepressant regimens, he was treated successfully with a course of bilateral electroconvulsive therapy. The patient experienced a robust response to treatment, with a 34 to 6 reduction in Hamilton Depression Rating Scale score. The potential challenges associated with administering electroconvulsive therapy to this patient population are discussed. PMID- 15791178 TI - Prolonged electroconvulsive therapy seizure in a patient taking ciprofloxacin. AB - Electroconvulsive therapy (ECT) can be used for the treatment of mental disorders, either alone or along with psychotropic agents and/or drugs for other medical conditions. We present in this case report a patient with postpartum depression on ciprofloxacin therapy for a urinary tract infection. The seizure in the first ECT treatment lasted for 30 seconds. While under the care of ECT, the patient was diagnosed as having a urinary tract infection; therefore, ciprofloxacin therapy of 1000 mg/day was initiated. Her second ECT seizure, which was on the third day of ciprofloxacin therapy, was terminated with 3 mg of intravenous midazolam at the 150th second. The ciprofloxacin therapy was discontinued. The patient had not previous history of epilepsy, and the investigation results for the extended seizure were found to be normal. The ECT therapy was restarted 3 days later, and total of 8 treatments were completed, lasting 35-70 seconds. Because the first ECT lasted for 30 seconds and subsequent therapy, which was reinitiated 3 days after the discontinuation of ciprofloxacin, lasted no longer than 70 seconds, the extended seizure in this patient is thought to be related to ciprofloxacin. PMID- 15791179 TI - Family member presence during electroconvulsive therapy: patient rights versus medical culture. PMID- 15791180 TI - Aspiration and postictal agitation after electroconvulsive therapy with propofol but no succinylcholine or atropinic agent. PMID- 15791181 TI - Fall in ECT use in young people in Edinburgh. PMID- 15791182 TI - First-line use of ECT. PMID- 15791183 TI - High-risk electroconvulsive therapy and patient autonomy: a response to Dr. McCall's editorial. PMID- 15791184 TI - Inflammation and depression: further studies are needed. PMID- 15791186 TI - [Surgical treatment of lumbosacral spondylolisthesis with major displacement in children and adolescents: a continuous series of 20 patients with mean 5-year follow-up]. AB - PURPOSE OF THE STUDY: This retrospective analysis involved a continuous series of twenty cases of lumbosacral spondylolisthesis with major displacement treated before maturity. We compared our experience using a non-instrumented lumbosacral fusion technique with the results of other techniques proposed in the literature. MATERIAL AND METHODS: We reviewed the cases of twenty children and adolescents who underwent surgery in our unit. For each case, we recorded the clinical history and course to last follow-up. X-rays were studied and manual measurements taken of the different parameters used to analyze the spine. Data were recorded in a database for statistical analysis. Sixteen of the twenty cases showed spinal deformation causing lumbalgia, generally associated with radiculalgia. Mean age at surgical treatment was 13 years 3 months, range 7 years 2 months to 17 years 6 months. All of the children has Meyerding stage 3 or 4 displacement associated with lumbosacral kyphosis. Surgical treatment followed a period of progressive reduction by traction and suspension in lordosis using a hammock. All twenty patients underwent posterolateral arthordesis using a cancellous graft between L4 and the sacrum. The fusion was performed after fashioning a thoraco-lumbo-pelvic cast including both thighs in the position of reduction. A complementary time for anterior arthodesis was needed for eight patients. RESULTS: The postoperative period was uneventful in twelve patients. Two children developed intestinal obstruction with a peritoneal bridle. Three children had an L5 radicular deficit and three sphincter disorders. All neurological disorders resolved in a few months. At mean postoperative follow-up of 5 years 3 months, the arthrodesis appeared to be fused in 19/20 cases. Only one patient presented a lucent line in the zone of the bone graft suggesting possible fibrous nonunion. Eighteen patients were symptom free and led a normal life. Two patients complained of moderately bothersome lower back pain. DISCUSSION: Many of the children in our series had major lumbosacral dysplasia with a verticalized sacrum, aggravating the lumbosacral kyphosis. This led to an increased pelvic tilt and decreased sacral slope. Progressive preoperative reduction of the lumbosacral kyphosis allowed conducting the lumbosacral fusion under favorable conditions. We did not open the spinal canal and avoided the mid line in order to protect as much as possible posterior spinal stability and preserve all the bone surfaces receiving the posterolateral graft. We reserved indications for complementary anterior lumbosacral arthrodesis to the most exaggerated cases of lumbosacral kyphosis. The therapeutic program is long due to the progressive preoperative reduction and the strict period of immobilization after surgery. In our experience, this approach allows quality lumbosacral fusion with good correction of the lumbosacral kyphosis. Neurological complications remain frequent and can occur during even slow progressive reduction. PMID- 15791188 TI - [Results of rheumatoid wrist surgery (arthrodesis excepted): 16 patients with more than 20 year follow-up]. AB - PURPOSE OF THE STUDY: The absence of a medical treatment capable of successfully arresting joint destruction due to rheumatoid arthritis (RA) leaves a large domain for surgical treatment. The purpose of our work was to determine whether a clinical benefit persists in the long term (more than 20 years) despite aggravation of the radiological lesions, after surgical treatment of rheumatoid arthritis of the wrist. MATERIAL AND METHODS: Sixteen patients with RA (13 women and 3 men, mean age 65 years), were reviewed a mean 24.8 years (range 20-33 years) after wrist surgery. Twenty-four wrists were operated for dorsal synovectomy (n=18) and Swanson radiocarpal implant (n=6). Total arthrodeses were excluded. Clinical, functional and standard and stress x-ray data were collected at last follow-up. RESULTS: Residual pain at last follow-up in wrists which had undergone dorsal synovectomy was scored 3.1/10 on the VAS versus 5.6 preoperatively. Three-quarters of the patients stated they were satisfied with the intervention despite very weak force. Revision surgery was required in eight patients after dorsal synovectomy including three which required resection of the ulnar head, left in place after the first surgery, and three for removal of a silicon implant of the ulnar head. This implant was rapidly abandoned in our unit (as in other units). The radiological status worsened in all wrists over time, despite synovectomy. For the Swanson radiocarpal implant, residual pain was only 0.5/10 versus 6.7 preoperatively. Four implants fractured and four developed radiological signs of siliconitis with not clinical expression. Despite these complications, five of the six patients felt favorably about their intervention and the mean Leclerc function score was 78/100. Flexion-extension was 56 degrees on average. The main complaint was the lack of force. CONCLUSION: There is a discordance between radiological and clinical results, a difference which widens with longer follow-up. A clear improvement in the pain score and the moderate functional demands of these patients are probably the reasons for their satisfaction despite radiological degradation. Many desire more wrist force. Our indications have evolved over time with the development after 1980 of the radiolunar arthrodesis procedures that we associate with dorsal synovectomy even in early-stage patients in order to limit radiological degradation and ulnar translation of the carpus. Swanson radiocarpal implants were completely abandoned in 1987 despite favorable clinical results due to the radiological degradation with bone loss and risk of siliconitis. For Simmen III wrists, total arthrodesis remains the only sure and definitive solution. PMID- 15791187 TI - [Twenty nine shoulder reconstructions after resection of the proximal humerus for neoplasm with mean 7-year follow-up]. AB - PURPOSE OF THE STUDY: Techniques available for shoulder reconstruction after resection of a tumor of the proximal humerus include scapulohumeral arthrodesis, humerus prosthesis with or without an allograft, inverted prostheses, and massive allografts. The purpose of this study was to review clinical and radiological outcomes in a series of 29 patients (20 men and 9 women) who underwent resection reconstruction of the proximal humerus and to establish from these cases a decision making algorithm for therapeutic indications as a function of tumor invasion. MATERIAL AND METHODS: The tumors were 20 chondrosarcomas, five osteosarcomas, two Ewing sarcomas and one malignant hemangiopericytoma. In 17 patients epiphyso-metaphyseal or epiphyso-metaphyso-diaphyseal resection was performed with preservation of the abductor muscles (type S34A or S345A according to the Musculoskeletal Tumor Society classification). For 12 patients epiphyso metaphyseal or epiphyso-metaphyso-diaphyseal resection was performed without preservation of the abductor muscles (type S34B or S345B). Reconstruction was achieved using a centromedullary cemented nail in one patient, scapulohumeral arthrodesis in three, a massive humerus prosthesis in 15, and composite humerus prosthesis in three and an inverted prosthesis in seven. The functional score of the Musculoskeletal Tumor Society (MSTS) and standard x-rays were used to assess outcome. RESULTS: Mean follow-up was 85 months (range 16-300). The mean MSTS score was 88% for inverted prostheses, 76% for composite prostheses, 72.6% for massive prostheses, 75% for scapulohumeral arthrodeses, 67% for massive prostheses, and 80% for cemented centromendullary nail. Five patients died from their malignant disease and one from another cause. Four patients are alive but with active disease after a mean follow-up of 108 months and 19 patients (65.5%) are alive and free of locoregional recurrence or metastasis after a mean 83.5 months. We had 28 complications. Glenohumeral instability was the most frequent (11 cases). DISCUSSION: Resection of the upper portion of the humerus should be performed to achieve cancerologically satisfactory tumor resection and enable shoulder resection, if possible, with preservation of a viable and functional abductor system. The functional outcome after such reconstruction depends on the type of bony resection, but also on the sacrifice of the rotator cuff and the deltoid muscle. In light of our experience and results in the literature, we advocate, despite the small number of cases for the different reconstructions, the following decision-making algorithm after resection of the proximal humerus without joint invasion: when the resection removes the rotator cuff and the deltoid (or the axillary nerve), there are two options: scapulohumeral arthrodesis or massive humerus prosthesis for patients who do not desire a complex therapy with a long postoperative period; when the resection preserves the rotator cuff and/or the deltoid muscle, reconstruction can be achieved with a composite (inverted or not) prosthesis with suture of the cuff tendons. We prefer the inverted composite prosthesis; if the deltoid muscle can be preserved but not the rotator cuff, the composite inverted prosthesis appears to be the most logical solution, but scapulohumeral arthrodesis can be proposed in selected cases. PMID- 15791189 TI - [Reconstruction of acute posterior cruciate ligament tears using a synthetic ligament]. AB - PURPOSE OF THE STUDY: Treatment of recent laxity of the posterior cruciate ligament is not standardized. The purpose of this work was to analyze results of reconstruction with adjunction of a synthetic ligament for major recent isolated or combined laxity of the posterior cruciate ligament (triades, pentades or dislocations). Our hypothesis was that the synthetic ligament acts like a tutor for healing of the torn ligament. MATERIAL AND METHODS: This retrospective analysis included 14 patients (1 woman and 13 men), mean age 27 years. All were competition athletes except one who did not practice sports. Three quarters of the patients were traffic accident victims. The series included three isolated posterior ligament tears, six combined laxities, and five knee dislocations. Average posterior laxity was 24 mm preoperatively. The procedure was performed 7 to 53 days after the accident. Arthroscopic reconstruction was performed for six patients and arthrotomy for eight. All associated lesions were repaired during the same procedure except for two cases (one anterior cruciate ligament and one popliteal tendon). Posterior cruciate ligament repair was achieved with the adjunction of a polyester ligament (LARS) using a one or two strand technique. Patients were reviewed at 36 months mean follow-up (10 - 88 months). The IKDC score was determined. A posterior drawer was measured manually with Telos at 70 degrees. RESULTS: Five stiff knees required either mobilization under anesthesia or arthrolysis. One tear occurred late after the accident during a new trauma. Subjectively, two patients were very satisfied, eight satisfied and three disappointed. Mean knee motion measurements were 6/0/130 degrees . A differential posterior drawer persisted in twelve knees. The Telos measurement of posterior drawer changed from a mean 24 mm to a mean 8 mm. The overall IKDC score was A: 0, B: 7, C: 3, and D: 2. Persistent posterior laxity was the predominant cause of poor scores. Outcome was less satisfactory for all items of posterolateral laxity. There was no difference between the 2- and 4-strand techniques. There were no cases of morbidity (synovitis, spontaneous tear) directly related to the synthetic ligament. DISCUSSION: The gain in posterior laxity was substantial. Results depended on associated lesions, particularly lateral involvement (stiffness, IKDC score) rather than the repair technique. The synthetic ligament appeared to play the role of a tutor: a single strand measuring 6 mm in diameter is sufficient. This technique spares tendon stock and could be proposed for major posterior cruciate ligament laxity. A longer follow-up will be necessary to confirm the durable stability. PMID- 15791190 TI - [Knee kinematics in flexion and extension using an electrogoniometric device: results in nine healthy subjects]. AB - PURPOSE OF THE STUDY: We wanted to develop a gait evaluation protocol using an electrogoniometric device allowing simpler measurements than optoelectronic recordings. MATERIAL AND METHODS: Gait was studied in a group of nine healthy subjects, mean age 58 years, walking on flat ground at three different speeds. We recorded the knee flexion and extension angles and the main temporospatial gait parameters. The principal conditions required for considering gait to be normal were taken from the literature. RESULTS: The subjects exhibited gait cycles which were normal in 83% of the cases for both knees at all three walking speeds. In 17% of the cases, the subjects walked with a flexed knee or without flexion during the weight-bearing phase. Knee motion was symmetrical between the right and left knee in all subjects. Increased walking speed was associated with increases temporospatial parameters and greater flexion and extension angles, particularly during the weight-bearing phase. There was a correlation between the weight-bearing flexion phase and stride length. DISCUSSION: The walking distance was moderate but sufficient to avoid the influence of the start and stop cycles. The electrogoniometers were small and did not perturb walking. Certain temporospatial parameters could not be measured directly but could be estimated. We checked that the results corresponded with data in the literature. Lateralization (right, left dominance) did not influence knee movement. Flexion during weight-bearing could influence gait quality by affecting the balance between gravity and quadriceps muscle force, particularly when considering forces used to progress forward. Knee motion varies with walking speed and depends not only on overall knee motion measured without weight bearing, but probably also on the interaction between joint kinematics, the musculoligamentary system, and the locomotor schema. PMID- 15791191 TI - [Osteolysis in revision total knee arthroplasty: a comparative study of osteolysis with and without metallosis]. AB - PURPOSE OF THE STUDY: Metallic debris from metallosis can compensate for the radiolucency resulting from bony destruction and compromise preoperative diagnosis of osteolysis. We compared the radiological findings in two groups of patients: those with pure osteolysis resulting from polyethylene debris and those with osteolysis associated with metallosis. MATERIAL AND METHODS: This study compared ten total knee arthroplasties free of metallosis at revision (pathology examination revealed only polyethylene debris) with ten total knee arthroplasties with pathologically confirmed metallosis of the synovium and periprosthetic bony tissue. Revisions were performed for loosening or a femoropatellar problem. At revision, the localization and the degree of tibial and/or femoral osteolysis were noted. Preoperative x-rays were analyzed to search for femoral, tibial or patellar osteolysis and were compared with operative findings. On the AP and lateral views, ten osteolysis zones were defined for each knee (four tibia, five femur, and one patella). RESULTS: Ten knees had osteolysis associated with polyethylene debris alone. Tibial osteolysis was found intraoperatively in 16 of the 40 theoretically possible localizations and was recognized on the preoperative x-rays in all 16. Femoral osteolysis was found intraoperatively in 18 of the 50 theoretically possible localizations was only recognized in 6 of the 18. For the ten knees with metallosis, tibial osteolysis was found intraoperatively in 22 of the theoretically possible localizations and was recognized on preoperative x-rays in 10 of the 22. Femoral osteolysis was found intraoperatively in 32 of the 50 theoretically possible localizations but was only recognized preoperatively in 5 of the 32. Comparisons between the two groups showed that femoral osteolysis was significantly more difficult to identify preoperatively than tibial osteolysis irrespective of the type of osteolysis (with or without metallosis). Furthermore, in patients with metallosis, it was significantly more difficult to recognize osteolysis preoperatively in both the tibial and femoral localizations. DISCUSSION: Metallosis can mask the diagnosis of femoral osteolysis and makes it very difficult to recognize tibial osteolysis. Arguments in favor of osteolysis (abnormal skin pigmentation, radio-opaque effusion, abnormal color and density of the joint fluid) are discussed. When the diagnosis of metallosis is established preoperatively, the chances of finding osteolysis intraoperatively are much higher than suggested by the preoperative x rays. PMID- 15791192 TI - [Ilizarov fixation of supramalleolar fractures]. AB - PURPOSE OF THE STUDY: Supramalleolar fractures are generally considered to be a difficult surgical challenge because they occur in a area where the tibia lies superficially with a precarious blood supply to the skin, exposing to the risk of infection and necrosis after internal fixation. These fractures are also situated close to the tibiotalar joint making centromedullary nailing difficult, even with distal locking. The Ilizarov external fixator could be an attractive alternative in this indication. MATERIAL AND METHODS: We report a series of 17 supramalleolar fractures in 17 patients, 14 men and 3 women, treated with the Ilizarov external fixator between 1991 and 2001. Most were traffic accident victims and most had complex fractures resulting from high-energy trauma. There were many associated lesions. Fractures were open in ten patients. The Ilizarov fixator was used as the first intention treatment in seven patients and as a second line treatment in ten. The system allowed early weight bearing in all patients. RESULTS: Tolerance was generally good with a relatively low rate of superficial pin track infections (two cases). There was one case of osteitis which developed in a patient with an open fracture. There were no thromboembolic complications and no nerve involvement. Bone healing was achieved within three months in thirteen patients. There were three cases of late healing which were treated by the ascension technique using a cancellous graft and fibular osteotomy. The overall healing rate with this method was 94%. The one case of nonunion was successfully treated with an inter tibiofibular graft. The overall functional outcome was satisfactory in 76% of the patients, based on the Alho-Klemm criteria. Axial deformation predominated in the frontal plane: three patients had > 10 degrees varus in one case. DISCUSSION: These results could be improved by better operative technique. We advocate installing the patient in the supine position with transcalcaneal traction allowing good restitution of the leg axis. The assembly should be long, including the entire leg segment. PMID- 15791194 TI - [Can metal-on-metal bearings improve the longevity of total hip prostheses?]. AB - PURPOSE OF THE STUDY: In total hip arthroplasty (THA), inter-series comparative clinical results cannot be considered worthy before at least 10 years of average follow-up, as shown in the Swedish Arthroplasty Register experience (1978-1993). Last generation metal-on-metal bearings were introduced in France only nine years ago (1995). To date, using mid term information, data from the literature, and our experience, one could assume that this bearing material has the capacity to improve THA longevity. Three types of information were analyzed: 1) comparative radiographic and EBRA studies published on early migration (<2 years) of acetabular implants; 2) preliminary comparative data on wear and osteolysis at the 8-year maximum follow-up; 3) clinical data on dislocation frequency and in vitro and in vivo observations of bearing separation and sliding. Study no 1: primary stability of these metal-on-metal acetabular cups was better than for polyethylene (PE) cups or alumina liners; study no 2: no detectable wear and less osteolysis were observed to date with metal-on-metal bearings versus their alumina-on-PE counterparts. Study no 3: less dislocation and less head sliding were measured with the metal-on-metal versus alumina-on-PR bearings possibly due to the interfacial forces provided by lubricating fluid (suction fit). CONCLUSION: According to current knowledge, this metal-on-metal bearing still represent, with alumina-on-alumina and highly crosslinked PE, a competitive and comprehensive option to improve THA longevity. The real pending problem concerns the frequency and above all the intensity of metal delayed hypersensitivity reaction and their potential effect on implant loosening. PMID- 15791193 TI - [Prevention of heterotopic ossifications in hip arthroplasty: effectiveness of selective Cox-2 inhibitors (celecoxib) versus ketoprofen]. AB - PURPOSE OF THE STUDY: Peri-prosthetic ossifications are a frequent complication of total hip arthroplasty, which, if effective preventive measures are not taken, occur in 60% of patients. Numerous classic antiinflammatory agents have shown their preventive efficacy. New selective Cox-2 inhibitors offer the possibility of reducing the adverse effects of antiinflammatory drugs but remain to be proven effective in this indication. The purpose of this pilot study was to compare the efficacy of celecoxib versus ketoprofen. MATERIAL AND METHODS: In order to obtain sufficient statistical power to have a 70% chance of detecting a 25% difference between the two treatments with a 5% risk of error, we evaluated the incidence of peri-prosthetic ossifications of the hip in a prospective monocentric series of 52 patients receiving 400 mg celecoxib a day during a week. It was compared with the incidence in a control series of 52 matched patients (same age, gender, diagnosis, operator experience) given 200 mg ketoprofen iv for 48 hr then 300 mg po for 5 days as preventive treatment. Ossifications were studied on the plain AP view of the pelvis at a mean follow-up of more than 11 months (11.4 vs 11.9). The Brooker classification was determined. RESULTS: The percent of patients presenting peri-prosthetic ossifications was equivalent. For the celecoxib group, 60% of the patients were free of ossifications; 28.9% presented stage 1 ossification and 11.1% stage 2 ossification; none of the hip exhibited a higher stage. In the ketoprofen control group, 53.2% of patients were free of ossification, 38.2% had stage 1 ossification, 6.4% stage 2, and 2.1% stage 3; there was no patient with stage 4. Fisher's exact test did not demonstrate a significant difference between the groups (p<0.51). Compared with an older series of patients who were not given preventive treatment, there was a significant reduction in incidence of peri-prosthetic ossification (p=0.014). DISCUSSION: The two study groups were not significantly different for age, gender, or underlying disease. There were an equivalent number of cases of intolerance to treatment in the two groups. CONCLUSION: These findings appear to indicate an equivalent efficacy for celecoxib and ketoprofen for the reduction of peri-prosthetic ossifications. Based on these results, a randomized prospective comparative study can be undertaken without risk of losing effective prevention in one group. This prospective study should enable a more precise evaluation of treatment equivalence and quantify any potential gain in morbidity obtained with celecoxib. PMID- 15791195 TI - [Adaptative femoroplasty in total hip arthroplasty for proximal femur deformity]. AB - Using a illustrative case, we describe a new surgical approach simplifying implantation of a total hip prosthesis for major proximal femur deformity. The technique uses femorotomy and osteotomy to short circuit the deformation with a non-cemented modular femoral stem with shaft anchorage allowing reconstruction of the proximal femur around the prosthesis, i.e. adaptative femoroplasty. PMID- 15791197 TI - Drawings, the universal language of children. The experience within a Department of Pediatric Hematology and Oncology. PMID- 15791198 TI - Alcohol use disorder in adolescents. AB - Alcohol use and abuse by children and adolescents remains a critical problem for modern developed countries. Although tolerance and public policy for alcohol use varies among Western countries, the use of alcohol and other harmful substances is common among adolescents. The use of alcohol can lead to a variety of negative consequences for youth. The risk for alcohol use and abuse, the acquisition of use behaviors, and development into alcohol use disorders and interventions for such problems should be considered in a comprehensive manner that considers neurobiology, development, and the adolescent's environmental ecology. Although the nature and extent of the alcohol problems between countries may vary, approaches to assessment and treatment from the United States may be useful to clinicians in other countries. This paper presents essential background information for the clinician in order to understand the presentation, risk, prevention and treatment of adolescents with alcohol use problems. PMID- 15791199 TI - [The pharmacological manipulation of Botallo's duct in the duct-dependent congenital cardiopathies and in the preterm infants with respiratory distress. A review and personal findings]. AB - Botallo's duct connects the systemic and pulmonary circulation. It can play a crucial hemodynamic role in some cardiac and respiratory diseases of the newborn, and strongly influences the outcome if it remains patent after birth or if it closes rapidly. The recently acquired in-depth knowledge on the genesis of these events have advanced the so called ''pharmacological manipulation of Botallo's duct'', i.e. pharmacological treatment to regulate the opening or closure of the ductus depending on clinical requirements and that, as will be described, is a key issue in managing newborns with severe cardiac and/or respiratory distress. This study illustrates the main underlying mechanisms of duct patency during intrauterine life and its closure after birth, and then describes the clinical conditions of the newborn where Botallo's duct must be kept patent after birth (duct-dependent cardiac malformations) and where its closure must be accelerated (patent duct associated idiopathic respiratory syndrome). It also reports the recent findings on the use of prostaglandins (PGE1) and prostaglandin synthesis inhibitors (indomethacin, ibuprofen) and the potential use of drugs capable of favouring or inhibiting nitric oxide in the duct endothelium. PMID- 15791200 TI - [Efficacy of toxin-A Botulinum for treating intractable bladder hyperactivity in children affected by neuropathic bladder secondary to myelomeningocele: an alternative to enterocystoplasty]. AB - AIM: To verify the efficacy of botulinus toxin A (TB-A) in treating children with neuropathic bladder secondary to myelomeningocele (MMC) with detrusor hyperactivity/low compliance, resistance to pharmacological therapy, and candidates for enterocystoplasty. METHODS: From January 2002 to June 2003, a group of 7 patients was selected (4 females, 3 males, mean age, 9.8 years, age range, 5-17 years) with detrusor hyperactivity, clean intermittent catheterization and resistance to pharmacological therapy. Two patients presented with grade 2-3 monolateral vesicoureteral reflux (VUR). All patients were incontinent despite catheterization. Botulinus toxin A was administered under general or local anesthesia by the injection of 200 IU of toxin diluted in 10 cc of physiologic solution with a metal or a flexible needle (3.7 F/21 GA). The needle was fully inserted into the detrusor muscle in about 20 sites, and 0.5 cc of solution were injected in each site, except the trigonum vesicae. Follow-up included ultrasound examination of the urinary tract and urodynamic studies performed at 6, 12 and 24 weeks and biannually thereafter. Micturition cystography was performed 3 months after the intervention. Urodynamic parameters were leak point pressure (LPP), leak point volume (LPV) and specific volume at 20 cm H2O pressure. The results were analyzed using the Wilcoxon test. RESULTS: A significant increase in LPV (range, 30-108%, mean, 77.6%) and in specific volume at 20 cm H2O pressure (CS 20) was observed in all patients. No significant change in LPP was found. One patient previously treated with the Cohen reimplantation technique experienced transient VUR which resolved spontaneously within 1 month. No major side effects from the injection of TB-A occurred. All patients were hospitalized for 24 hours with catheterization. CONCLUSIONS: The preliminary results in this small sample of patients suggest that the use of TB-A is efficacious in significantly improving urodynamic parameters and urine storage volume at low pressures in patients with neuropathic bladder resistant to pharmacological therapy. PMID- 15791201 TI - [Height growth velocity and orthodontic treatment timing]. AB - The auxologic evaluation, by periodic measurements of the height of children and the comparison of the values obtained with standard curves of growth, is a common practice for the pediatrician providing a precise indication on the general course of the growth and on the presence of possible growth pathologies. In fact, decelerations or pathological accelerations of growth can be a suspicious sign of endocrine, metabolic, genetic, gastric or tumoral diseases. The use of some growth variables and particularly the evaluation of the kinetics of statural height growth, should enter the daily practice of the orthodontist. In fact, the correspondence among peaks of statural and mandibular growth allows the orthodontist to choose the suitable time to intervene with the therapy aimed to stimulate the growth of the jaws. In this paper the authors report 2 orthodontic cases characterized by a malocclusion due to mandibular deficiency, in which the integration of the orthodontic and pediatric competences, through the use of the curves of the speed of growth and other growth variables, has allowed to reach optimal results, having achieved normal skeletal and occlusal relationships in the shorter time. The periodic measurement of height growth velocity represents a simple and safe method that allows to evaluate the kinetics of growth in children, with the remarkable advantage, unlike radiological examinations, to be repeated without any risk for the patient. PMID- 15791202 TI - [Are holidays always safe: immigrants and hepatitis A]. PMID- 15791203 TI - Back to sleep: a longitudinal report from an infant population based study in the Local Health Service 11, Piedmont, Italy. PMID- 15791204 TI - Recoding elements located adjacent to a subset of eukaryal selenocysteine specifying UGA codons. AB - Incorporation of the 21st amino acid, selenocysteine, into proteins is specified in all three domains of life by dynamic translational redefinition of UGA codons. In eukarya and archaea, selenocysteine insertion requires a cis-acting selenocysteine insertion sequence (SECIS) usually located in the 3'UTR of selenoprotein mRNAs. Here we present comparative sequence analysis and experimental data supporting the presence of a second stop codon redefinition element located adjacent to a selenocysteine-encoding UGA codon in the eukaryal gene, SEPN1. This element is sufficient to stimulate high-level (6%) translational redefinition of the SEPN1 UGA codon in human cells. Readthrough levels further increased to 12% when tested in the presence of the SEPN1 3'UTR SECIS. Directed mutagenesis and phylogeny of the sequence context strongly supports the importance of a stem loop starting six nucleotides 3' of the UGA codon. Sequences capable of forming strong RNA structures were also identified 3' adjacent to, or near, selenocysteine-encoding UGA codons in the Sps2, SelH, SelO, and SelT selenoprotein genes. PMID- 15791205 TI - Receptor and viral determinants of SARS-coronavirus adaptation to human ACE2. AB - Human angiotensin-converting enzyme 2 (ACE2) is a functional receptor for SARS coronavirus (SARS-CoV). Here we identify the SARS-CoV spike (S)-protein-binding site on ACE2. We also compare S proteins of SARS-CoV isolated during the 2002 2003 SARS outbreak and during the much less severe 2003-2004 outbreak, and from palm civets, a possible source of SARS-CoV found in humans. All three S proteins bound to and utilized palm-civet ACE2 efficiently, but the latter two S proteins utilized human ACE2 markedly less efficiently than did the S protein obtained during the earlier human outbreak. The lower affinity of these S proteins could be complemented by altering specific residues within the S-protein-binding site of human ACE2 to those of civet ACE2, or by altering S-protein residues 479 and 487 to residues conserved during the 2002-2003 outbreak. Collectively, these data describe molecular interactions important to the adaptation of SARS-CoV to human cells, and provide insight into the severity of the 2002-2003 SARS epidemic. PMID- 15791206 TI - Role that phosphorylation of GSK3 plays in insulin and Wnt signalling defined by knockin analysis. AB - The inactivation of glycogen synthase kinase (GSK)3 has been proposed to play important roles in insulin and Wnt signalling. To define the role that inactivation of GSK3 plays, we generated homozygous knockin mice in which the protein kinase B phosphorylation sites on GSK3alpha (Ser21) and GSK3beta (Ser9) were changed to Ala. The knockin mice were viable and were not diabetic. Using these mice we show that inactivation of GSK3beta rather than GSK3alpha is the major route by which insulin activates muscle glycogen synthase. In contrast, we demonstrate that the activation of muscle glycogen synthase by contraction, the stimulation of muscle glucose uptake by insulin, or the activation of hepatic glycogen synthase by glucose do not require GSK3 phosphorylation on Ser21/Ser9. GSK3 also becomes inhibited in the Wnt-signalling pathway, by a poorly defined mechanism. In GSK3alpha/GSK3beta homozygous knockin cells, Wnt3a induces normal inactivation of GSK3, as judged by the stabilisation of beta-catenin and stimulation of Wnt-dependent transcription. These results establish the function of Ser21/Ser9 phosphorylation in several processes in which GSK3 inactivation has previously been implicated. PMID- 15791207 TI - Structure of P-protein of the glycine cleavage system: implications for nonketotic hyperglycinemia. AB - The crystal structure of the P-protein of the glycine cleavage system from Thermus thermophilus HB8 has been determined. This is the first reported crystal structure of a P-protein, and it reveals that P-proteins do not involve the alpha(2)-type active dimer universally observed in the evolutionarily related pyridoxal 5'-phosphate (PLP)-dependent enzymes. Instead, novel alphabeta-type dimers associate to form an alpha(2)beta(2) tetramer, where the alpha- and beta subunits are structurally similar and appear to have arisen by gene duplication and subsequent divergence with a loss of one active site. The binding of PLP to the apoenzyme induces large open-closed conformational changes, with residues moving up to 13.5 A. The structure of the complex formed by the holoenzyme bound to an inhibitor, (aminooxy)acetate, suggests residues that may be responsible for substrate recognition. The molecular surface around the lipoamide-binding channel shows conservation of positively charged residues, which are possibly involved in complex formation with the H-protein. These results provide insights into the molecular basis of nonketotic hyperglycinemia. PMID- 15791208 TI - Role of curvature and phase transition in lipid sorting and fission of membrane tubules. AB - We have recently developed a minimal system for generating long tubular nanostructures that resemble tubes observed in vivo with biological membranes. Here, we studied membrane tube pulling in ternary mixtures of sphingomyelin, phosphatidylcholine and cholesterol. Two salient results emerged: the lipid composition is significantly different in the tubes and in the vesicles; tube fission is observed when phase separation is generated in the tubes. This shows that lipid sorting may depend critically on both membrane curvature and phase separation. Phase separation also appears to be important for membrane fission in tubes pulled out of giant liposomes or purified Golgi membranes. PMID- 15791210 TI - Endoplasmic reticulum BIK initiates DRP1-regulated remodelling of mitochondrial cristae during apoptosis. AB - The endoplasmic reticulum (ER) can elicit proapoptotic signalling that results in transmission of Ca(2+) to the mitochondria, which in turn stimulates recruitment of the fission enzyme DRP1 to the surface of the organelle. Here, we show that BH3-only BIK activates this pathway at the ER in intact cells, resulting in mitochondrial fragmentation but little release of cytochrome c to the cytosol. The BIK-induced transformations in mitochondria are dynamic in nature and involve DRP1-dependent remodelling and opening of cristae, where the major stores of cytochrome c reside. This novel function for DRP1 is distinct from its recognized role in regulating mitochondrial fission. Selective permeabilization of the outer membrane with digitonin confirmed that BIK stimulation results in mobilization of intramitochondrial cytochrome c. Of note, BIK can cooperate with a weak BH3-only protein that targets mitochondria, such as NOXA, to activate BAX by a mechanism that is independent of DRP1 enzyme activity. When expressed together, BIK and NOXA cause rapid release of mobilized cytochrome c and activation of caspases. PMID- 15791209 TI - Crystal structure of a Cbtx-AChBP complex reveals essential interactions between snake alpha-neurotoxins and nicotinic receptors. AB - The crystal structure of the snake long alpha-neurotoxin, alpha-cobratoxin, bound to the pentameric acetylcholine-binding protein (AChBP) from Lymnaea stagnalis, was solved from good quality density maps despite a 4.2 A overall resolution. The structure unambiguously reveals the positions and orientations of all five three fingered toxin molecules inserted at the AChBP subunit interfaces and the conformational changes associated with toxin binding. AChBP loops C and F that border the ligand-binding pocket move markedly from their original positions to wrap around the tips of the toxin first and second fingers and part of its C terminus, while rearrangements also occur in the toxin fingers. At the interface of the complex, major interactions involve aromatic and aliphatic side chains within the AChBP binding pocket and, at the buried tip of the toxin second finger, conserved Phe and Arg residues that partially mimic a bound agonist molecule. Hence this structure, in revealing a distinctive and unpredicted conformation of the toxin-bound AChBP molecule, provides a lead template resembling a resting state conformation of the nicotinic receptor and for understanding selectivity of curaremimetic alpha-neurotoxins for the various receptor species. PMID- 15791212 TI - Joys of (top-notch) supervision. PMID- 15791211 TI - Interaction of HSP90 to N-WASP leads to activation and protection from proteasome dependent degradation. AB - Neural Wiskott-Aldrich syndrome protein (N-WASP) regulates reorganization of the actin cytoskeleton through activation of the Arp2/3 complex. Here, we show that heat shock protein 90 (HSP90) regulates N-WASP-induced actin polymerization in cooperation with phosphorylation of N-WASP. HSP90 binds directly to N-WASP, but binding alone does not affect the rate of N-WASP/Arp2/3 complex-induced in vitro actin polymerization. An Src family tyrosine kinase, v-Src, phosphorylates and activates N-WASP. HSP90 increases the phosphorylation of N-WASP by v-Src, leading to enhanced N-WASP-dependent actin polymerization. In addition, HSP90 protects phosphorylated and activated N-WASP from proteasome-dependent degradation, resulting in amplification of N-WASP-dependent actin polymerization. Association between HSP90 and N-WASP is increased in proportion to activation of N-WASP by phosphorylation. HSP90 is colocalized and associated with active N-WASP at podosomes in 3Y1/v-Src cells and at growing neurites in PC12 cells, whose actin structures are clearly inhibited by blocking the binding of HSP90 to N-WASP. These findings suggest that HSP90 induces efficient activation of N-WASP downstream of phosphorylation signal by Src family kinases and is critical for N WASP-dependent podosome formation and neurite extension. PMID- 15791213 TI - US launches probe into sales of unapproved transgenic corn. PMID- 15791214 TI - No-confidence vote fails to shift Harvard president. PMID- 15791215 TI - Obesity expert owns up to million-dollar crime. PMID- 15791216 TI - Surfeit of boys could spread AIDS in China's cities. PMID- 15791217 TI - France takes on Google in scanning race. PMID- 15791218 TI - US undervalues foreign researchers, survey reveals. PMID- 15791219 TI - Academies seek better prospects for postdocs. PMID- 15791220 TI - Fake papers hamper plans for nuclear store at Yucca Mountain. PMID- 15791221 TI - Lawsuits and logistics tie up California's stem-cell funds. PMID- 15791223 TI - Condensed matter physics: some like it cold. PMID- 15791224 TI - Palaeoanthropology: looking for the ancestors. PMID- 15791227 TI - Consumer law is used to attack climate findings. PMID- 15791228 TI - Immigration could ease climate-change impact. PMID- 15791234 TI - Genetics: hotheaded healer. PMID- 15791235 TI - Genomics: frontiers of gene function. PMID- 15791236 TI - Earth science: a different kind of foreshock. PMID- 15791238 TI - Condensed-matter physics: lab in a trap. PMID- 15791237 TI - Physiology: do neural signals remodel bone? PMID- 15791239 TI - Biodiversity: gut feeling for yeasts. PMID- 15791241 TI - Nanotechnology: new spin on correlated electrons. PMID- 15791242 TI - Obituary: Hubert Curien (1924-2005). PMID- 15791244 TI - Animal behaviour: elephants are capable of vocal learning. AB - There are a few mammalian species that can modify their vocalizations in response to auditory experience--for example, some marine mammals use vocal imitation for reproductive advertisement, as birds sometimes do. Here we describe two examples of vocal imitation by African savannah elephants, Loxodonta africana, a terrestrial mammal that lives in a complex fission-fusion society. Our findings favour a role for vocal imitation that has already been proposed for primates, birds, bats and marine mammals: it is a useful form of acoustic communication that helps to maintain individual-specific bonds within changing social groupings. PMID- 15791245 TI - Meteorology: dusty ice clouds over Alaska. AB - Particles lofted into the atmosphere by desert dust storms can disperse widely and affect climate directly through aerosol scattering and absorption. They can also affect it indirectly by changing the scattering properties of clouds and, because desert dusts are particularly active ice-forming agents, by affecting the formation and thermodynamic phase of clouds. Here I show that dust storms that occurred in Asia early in 2004 created unusual ice clouds over Alaska at temperatures far warmer than those expected for normal cirrus-cloud formation. PMID- 15791246 TI - Foreshock sequences and short-term earthquake predictability on East Pacific Rise transform faults. AB - East Pacific Rise transform faults are characterized by high slip rates (more than ten centimetres a year), predominantly aseismic slip and maximum earthquake magnitudes of about 6.5. Using recordings from a hydroacoustic array deployed by the National Oceanic and Atmospheric Administration, we show here that East Pacific Rise transform faults also have a low number of aftershocks and high foreshock rates compared to continental strike-slip faults. The high ratio of foreshocks to aftershocks implies that such transform-fault seismicity cannot be explained by seismic triggering models in which there is no fundamental distinction between foreshocks, mainshocks and aftershocks. The foreshock sequences on East Pacific Rise transform faults can be used to predict (retrospectively) earthquakes of magnitude 5.4 or greater, in narrow spatial and temporal windows and with a high probability gain. The predictability of such transform earthquakes is consistent with a model in which slow slip transients trigger earthquakes, enrich their low-frequency radiation and accommodate much of the aseismic plate motion. PMID- 15791247 TI - Full-genome RNAi profiling of early embryogenesis in Caenorhabditis elegans. AB - A key challenge of functional genomics today is to generate well-annotated data sets that can be interpreted across different platforms and technologies. Large scale functional genomics data often fail to connect to standard experimental approaches of gene characterization in individual laboratories. Furthermore, a lack of universal annotation standards for phenotypic data sets makes it difficult to compare different screening approaches. Here we address this problem in a screen designed to identify all genes required for the first two rounds of cell division in the Caenorhabditis elegans embryo. We used RNA-mediated interference to target 98% of all genes predicted in the C. elegans genome in combination with differential interference contrast time-lapse microscopy. Through systematic annotation of the resulting movies, we developed a phenotypic profiling system, which shows high correlation with cellular processes and biochemical pathways, thus enabling us to predict new functions for previously uncharacterized genes. PMID- 15791248 TI - Filamentary structure on the Sun from the magnetic Rayleigh-Taylor instability. AB - Magnetic flux emerges from the solar surface as dark filaments connecting small sunspots with opposite polarities. The regions around the dark filaments are often bright in X-rays and are associated with jets. This implies plasma heating and acceleration, which are important for coronal heating. Previous two dimensional simulations of such regions showed that magnetic reconnection between the coronal magnetic field and the emerging flux produced X-ray jets and flares, but left unresolved the origin of filamentary structure and the intermittent nature of the heating. Here we report three-dimensional simulations of emerging flux showing that the filamentary structure arises spontaneously from the magnetic Rayleigh-Taylor instability, contrary to the previous view that the dark filaments are isolated bundles of magnetic field that rise from the photosphere carrying the dense gas. As a result of the magnetic Rayleigh-Taylor instability, thin current sheets are formed in the emerging flux, and magnetic reconnection occurs between emerging flux and the pre-existing coronal field in a spatially intermittent way. This explains naturally the intermittent nature of coronal heating and the patchy brightenings in solar flares. PMID- 15791249 TI - Spatial quantum noise interferometry in expanding ultracold atom clouds. AB - In a pioneering experiment, Hanbury Brown and Twiss (HBT) demonstrated that noise correlations could be used to probe the properties of a (bosonic) particle source through quantum statistics; the effect relies on quantum interference between possible detection paths for two indistinguishable particles. HBT correlations- together with their fermionic counterparts--find numerous applications, ranging from quantum optics to nuclear and elementary particle physics. Spatial HBT interferometry has been suggested as a means to probe hidden order in strongly correlated phases of ultracold atoms. Here we report such a measurement on the Mott insulator phase of a rubidium Bose gas as it is released from an optical lattice trap. We show that strong periodic quantum correlations exist between density fluctuations in the expanding atom cloud. These spatial correlations reflect the underlying ordering in the lattice, and find a natural interpretation in terms of a multiple-wave HBT interference effect. The method should provide a useful tool for identifying complex quantum phases of ultracold bosonic and fermionic atoms. PMID- 15791250 TI - Orbital Kondo effect in carbon nanotubes. AB - Progress in the fabrication of nanometre-scale electronic devices is opening new opportunities to uncover deeper aspects of the Kondo effect--a characteristic phenomenon in the physics of strongly correlated electrons. Artificial single impurity Kondo systems have been realized in various nanostructures, including semiconductor quantum dots, carbon nanotubes and individual molecules. The Kondo effect is usually regarded as a spin-related phenomenon, namely the coherent exchange of the spin between a localized state and a Fermi sea of delocalized electrons. In principle, however, the role of the spin could be replaced by other degrees of freedom, such as an orbital quantum number. Here we show that the unique electronic structure of carbon nanotubes enables the observation of a purely orbital Kondo effect. We use a magnetic field to tune spin-polarized states into orbital degeneracy and conclude that the orbital quantum number is conserved during tunnelling. When orbital and spin degeneracies are present simultaneously, we observe a strongly enhanced Kondo effect, with a multiple splitting of the Kondo resonance at finite field and predicted to obey a so called SU4 symmetry. PMID- 15791251 TI - Compact, stable and efficient all-fibre gas cells using hollow-core photonic crystal fibres. AB - Gas-phase materials are used in a variety of laser-based applications--for example, in high-precision frequency measurement, quantum optics and nonlinear optics. Their full potential has however not been realized because of the lack of a suitable technology for creating gas cells that can guide light over long lengths in a single transverse mode while still offering a high level of integration in a practical and compact set-up or device. As a result, solid-phase materials are still often favoured, even when their performance compares unfavourably with gas-phase systems. Here we report the development of all-fibre gas cells that meet these challenges. Our structures are based on gas-filled hollow-core photonic crystal fibres, in which we have recently demonstrated substantially enhanced stimulated Raman scattering, and which exhibit high performance, excellent long-term pressure stability and ease of use. To illustrate the practical potential of these structures, we report two different devices: a hydrogen-filled cell for efficient generation of rotational Raman scattering using only quasi-continuous-wave laser pulses; and acetylene-filled cells, which we use for absolute frequency-locking of diode lasers with very high signal-to-noise ratios. The stable performance of these compact gas-phase devices could permit, for example, gas-phase laser devices incorporated in a 'credit card' or even in a laser pointer. PMID- 15791252 TI - Obliquity pacing of the late Pleistocene glacial terminations. AB - The 100,000-year timescale in the glacial/interglacial cycles of the late Pleistocene epoch (the past approximately 700,000 years) is commonly attributed to control by variations in the Earth's orbit. This hypothesis has inspired models that depend on the Earth's obliquity (approximately 40,000 yr; approximately 40 kyr), orbital eccentricity (approximately 100 kyr) and precessional (approximately 20 kyr) fluctuations, with the emphasis usually on eccentricity and precessional forcing. According to a contrasting hypothesis, the glacial cycles arise primarily because of random internal climate variability. Taking these two perspectives together, there are currently more than thirty different models of the seven late-Pleistocene glacial cycles. Here we present a statistical test of the orbital forcing hypothesis, focusing on the rapid deglaciation events known as terminations. According to our analysis, the null hypothesis that glacial terminations are independent of obliquity can be rejected at the 5% significance level, whereas the corresponding null hypotheses for eccentricity and precession cannot be rejected. The simplest inference consistent with the test results is that the ice sheets terminated every second or third obliquity cycle at times of high obliquity, similar to the original proposal by Milankovitch. We also present simple stochastic and deterministic models that describe the timing of the late-Pleistocene glacial terminations purely in terms of obliquity forcing. PMID- 15791253 TI - Two episodes of microbial change coupled with Permo/Triassic faunal mass extinction. AB - Microbial expansion following faunal mass extinctions in Earth history can be studied by petrographic examination of microbialites (microbial crusts) or well preserved organic-walled microbes. However, where preservation is poor, quantification of microbial communities can be problematic. We have circumvented this problem by adopting a lipid biomarker-based approach to evaluate microbial community changes across the Permo/Triassic (P/Tr) boundary at Meishan in South China. We present here a biomarker stratigraphic record showing episodic microbial changes coupled with a high-resolution record of invertebrate mass extinction. Variation in the microbial community structure is characterized by the 2-methylhopane (2-MHP) index (a ratio of the abundance of cyanobacterial biomarkers to more general bacterial biomarkers). Two episodes of faunal mass extinction were each preceded by minima in the 2-MHP index, followed by strong maxima, likely reflecting microbial responses to the catastrophic events that caused the extinction and initiated ecosystem changes. Hence, both cyanobacterial biomarker and invertebrate fossil records provide evidence for two episodes of biotic crisis across the P/Tr boundary. PMID- 15791254 TI - Affinities of 'hyopsodontids' to elephant shrews and a Holarctic origin of Afrotheria. AB - Macroscelideans (elephant shrews or sengis) are small-bodied (25-540 g), cursorial (running) and saltatorial (jumping), insectivorous and omnivorous placental mammals represented by at least 15 extant African species classified in four genera. Macroscelidea is one of several morphologically diverse but predominantly African placental orders classified in the superorder Afrotheria by molecular phylogeneticists. The distribution of modern afrotheres, in combination with a basal position for Afrotheria within Placentalia and molecular divergence time estimates, has been used to link placental diversification with the mid Cretaceous separation of South America and Africa. Morphological phylogenetic analyses do not support Afrotheria and the fossil record favours a northern origin of Placentalia. Here we describe fossil postcrania that provide evidence for a close relationship between North American Palaeocene-Eocene apheliscine 'hyopsodontid' 'condylarths' (early ungulates or hoofed mammals) and extant Macroscelidea. Apheliscine postcranial morphology is consistent with a relationship to other ungulate-like afrotheres (Hyracoidea, Proboscidea) but does not provide support for a monophyletic Afrotheria. As the oldest record of an afrothere clade, identification of macroscelidean relatives in the North American Palaeocene argues against an African origin for Afrotheria, weakening support for linking placental diversification to the break-up of Gondwana. PMID- 15791255 TI - Evidence that sensory traps can evolve into honest signals. AB - Conventional models explaining extreme sexual ornaments propose that these reflect male genetic quality or are arbitrary results of genetic linkage between female preference and the ornament. The chase-away model emphasizes sexual conflict: male signals attract females because they exploit receiver biases. As males gain control of mating decisions, females may experience fitness costs through suboptimal mating rates or post-copulatory exploitation. Elaboration of male signals is expected if females increase their response threshold to resist such exploitation. If ornaments target otherwise adaptive biases such as feeding responses, selection on females might eventually separate sexual and non-sexual responses to the signal. Here we show that the terminal yellow band (TYB) of several Goodeinae species evokes both feeding and sexual responses; sexual responsiveness phylogenetically pre-dates the expression of the TYB in males and is comparable across taxa, yet feeding responsiveness decreases in species with more elaborated TYBs. Displaying a TYB is costly, and thus provides an example where a trait arose as a sensory trap but has evolved into an honest signal. PMID- 15791256 TI - Independent recruitment of a conserved developmental mechanism during leaf evolution. AB - Vascular plants evolved in the Middle to Late Silurian period, about 420 million years ago. The fossil record indicates that these primitive plants had branched stems with sporangia but no leaves. Leaf-like lateral outgrowths subsequently evolved on at least two independent occasions. In extant plants, these events are represented by microphyllous leaves in lycophytes (clubmosses, spikemosses and quillworts) and megaphyllous leaves in euphyllophytes (ferns, gymnosperms and angiosperms). Our current understanding of how leaves develop is restricted to processes that operate during megaphyll formation. Because microphylls and megaphylls evolved independently, different mechanisms might be required for leaf formation. Here we show that this is not so. Gene expression data from a microphyllous lycophyte, phylogenetic analyses, and a cross-species complementation experiment all show that a common developmental mechanism can underpin both microphyll and megaphyll formation. We propose that this mechanism might have operated originally in the context of primitive plant apices to facilitate bifurcation. Recruitment of this pathway to form leaves occurred independently and in parallel in different plant lineages. PMID- 15791257 TI - Recognition of bacterial glycosphingolipids by natural killer T cells. AB - Natural killer T (NKT) cells constitute a highly conserved T lymphocyte subpopulation that has the potential to regulate many types of immune responses through the rapid secretion of cytokines. NKT cells recognize glycolipids presented by CD1d, a class I-like antigen-presenting molecule. They have an invariant T-cell antigen receptor (TCR) alpha-chain, but whether this invariant TCR recognizes microbial antigens is still controversial. Here we show that most mouse and human NKT cells recognize glycosphingolipids from Sphingomonas, Gram negative bacteria that do not contain lipopolysaccharide. NKT cells are activated in vivo after exposure to these bacterial antigens or bacteria, and mice that lack NKT cells have a marked defect in the clearance of Sphingomonas from the liver. These data suggest that NKT cells are T lymphocytes that provide an innate type immune response to certain microorganisms through recognition by their antigen receptor, and that they might be useful in providing protection from bacteria that cannot be detected by pattern recognition receptors such as Toll like receptor 4. PMID- 15791258 TI - Exogenous and endogenous glycolipid antigens activate NKT cells during microbial infections. AB - CD1d-restricted natural killer T (NKT) cells are innate-like lymphocytes that express a conserved T-cell receptor and contribute to host defence against various microbial pathogens. However, their target lipid antigens have remained elusive. Here we report evidence for microbial, antigen-specific activation of NKT cells against Gram-negative, lipopolysaccharide (LPS)-negative alpha Proteobacteria such as Ehrlichia muris and Sphingomonas capsulata. We have identified glycosylceramides from the cell wall of Sphingomonas that serve as direct targets for mouse and human NKT cells, controlling both septic shock reaction and bacterial clearance in infected mice. In contrast, Gram-negative, LPS-positive Salmonella typhimurium activates NKT cells through the recognition of an endogenous lysosomal glycosphingolipid, iGb3, presented by LPS-activated dendritic cells. These findings identify two novel antigenic targets of NKT cells in antimicrobial defence, and show that glycosylceramides are an alternative to LPS for innate recognition of the Gram-negative, LPS-negative bacterial cell wall. PMID- 15791259 TI - Fission yeast Mes1p ensures the onset of meiosis II by blocking degradation of cyclin Cdc13p. AB - Meiosis is a special form of nuclear division to generate eggs, sperm and spores in eukaryotes. Meiosis consists of the first (MI) and the second (MII) meiotic divisions, which occur consecutively. MI is reductional, in which homologous chromosomes derived from parents segregate. MI is supported by an elaborate mechanism involving meiosis-specific cohesin and its protector. MII is equational, in which replicated sister-chromatids separate as in mitosis. MII is generally considered to mimic mitosis in mechanism. However, fission yeast Mes1p is essential for MII but dispensable for mitosis. The mes1-B44 mutant arrests before MII. Transcription of mes1 is low in vegetative cells and boosted in a narrow window between late MI and late MII. The mes1 mRNA undergoes meiosis specific splicing. Here we show that Mes1p is a factor that suppresses the degradation of cyclin Cdc13p at anaphase I. Mes1p binds to Slp1p, an activator of APC/C (anaphase promoting complex/cyclosome), and counteracts its function to engage Cdc13p in proteolysis. Inhibition of APC/C-dependent degradation of Cdc13p by Mes1p was reproduced in a Xenopus egg extract. We therefore propose that Mes1p has a key function in saving a sufficient level of MPF (M-phase-promoting factor) activity required for the execution of MII. PMID- 15791261 TI - Tale of two surveys. PMID- 15791260 TI - Recruitment of Drosophila Polycomb group proteins to chromatin by DSP1. AB - Polycomb and trithorax group (PcG and trxG) proteins maintain silent and active transcriptional states, respectively, throughout development. In Drosophila, PcG and trxG proteins associate with DNA regions named Polycomb and trithorax response elements (PRE and TRE), but the mechanisms of recruitment are unknown. We previously characterized a minimal element from the regulatory region of the Abdominal-B gene, termed Ab-Fab. Ab-Fab contains a PRE and a TRE and is able to maintain repressed or active chromatin states during development. Here we show that the Dorsal switch protein 1 (DSP1), a Drosophila HMGB2 homologue, binds to a sequence present within Ab-Fab and in other characterized PREs. Addition of this motif to an artificial sequence containing Pleiohomeotic and GAGA factor consensus sites is sufficient for PcG protein recruitment in vivo. Mutations that abolish DSP1 binding to Ab-Fab and to a PRE from the engrailed locus lead to loss of PcG protein binding, loss of silencing, and switching of these PREs into constitutive TREs. The binding of DSP1 to PREs is therefore important for the recruitment of PcG proteins. PMID- 15791262 TI - Pity poor postdocs. PMID- 15791264 TI - Recruiters and academia. Hard times for Swedish physician-researchers. PMID- 15791267 TI - Is yeast on its way to evolving tRNA editing? AB - In human mitochondria, genes for tRNA(Tyr) and tRNA(Cys) overlap by a single nucleotide. From polycistronic precursors, a 3'-truncated upstream tRNA(Tyr) is released, missing the overlapping position. A subsequent editing reaction restores this position. Similar mitochondrial tRNA gene overlaps exist in all metazoans, but not in organisms such as yeast or Escherichia coli. Therefore, we asked whether tRNA overlaps are processed in these organisms. Corresponding constructs were introduced and transcripts tested for processing and editing in E. coli and yeast. E. coli produces only one functional tRNA from these precursors, indicating that tRNA overlaps are incompatible with its processing pathway. In contrast, yeast processes overlapping tRNAs similar to human mitochondria, releasing a 3'-truncated upstream tRNA. This tRNA is restored in an editing-like event, although yeast does not carry a corresponding endogenous editing substrate. These findings support the hypothesis of the evolution of editing by recruitment of a pre-existing and promiscuous editing enzyme. PMID- 15791268 TI - The biology and biochemistry of diacylglycerol signalling. Meeting on molecular advances in diacylglycerol signalling. PMID- 15791269 TI - mPER1-mediated nuclear export of mCRY1/2 is an important element in establishing circadian rhythm. AB - Receptor-mediated nucleocytoplasmic transport of clock proteins is an important, conserved element of the core mechanism for circadian rhythmicity. A systematic analysis of the nuclear export characteristics for the different murine period (mPER) and cryptochrome (mCRY) proteins using Xenopus oocytes as an experimental system demonstrates that all three mPER proteins, but neither mCRY1 nor mCRY2, are exported if injected individually. However, nuclear injection of heterodimeric complexes that contain combinations of mPER and mCRY proteins shows that mPER1 serves as an export adaptor for mCRY1 and mCRY2. Functional analysis of dominant-negative mPER1 variants designed either to sequester mPER3 to the cytoplasm or to inhibit nuclear export of mCRY1/2 in synchronized, stably transfected fibroblasts suggests that mPER1-mediated export of mCRY1/2 defines an important new element of the core clock machinery in vertebrates. PMID- 15791272 TI - Home-based neonatal care: summary and applications of the field trial in rural Gadchiroli, India (1993 to 2003). AB - High levels of neonatal mortality and lack of access to neonatal health care are widespread problems in developing countries. A field trial of home-based neonatal care (HBNC) was conducted in rural Gadchiroli, India to develop and test the feasibility of a low-cost approach of delivering primary neonatal care by using the human potential available in villages, and to evaluate its effect on neonatal mortality. In the first half of this article we summarize various aspects of the field trial, presented in the previous 11 articles in this issue of the journal supplement. The background, objectives, study design and interventions in the field trial and the results over 10 years (1993 to 2003) are presented. Based on these results, the hypotheses are tested and conclusions presented. In the second half, we discuss the next questions: can it be replicated? Can this intervention become a part of primary health-care services? What is the cost and the cost effectiveness of HBNC? The limitations of the approach, the settings where HBNC might be relevant and the management pre-requisites for its scaling up are also discussed. The need to develop an integrated approach is emphasized. A case for newborn care in the community is made for achieving equity in health care. PMID- 15791273 TI - Methods and the baseline situation in the field trial of home-based neonatal care in Gadchiroli, India. AB - In this paper, we describe the planning of the field trial and the methods used for collecting baseline health and ethnographic data in a rural field study site. We describe the study hypotheses, specific objectives, study design, sample size estimates, selection of study area, community consent, the organization of study teams, review mechanism, financial support and baseline data collection. Baseline population characteristics and vital statistics are presented. The qualitative information on traditional beliefs and practices prevalent in the study area revealed that parents felt powerless about newborn health and sickness. There was an enormous unmet need to reach the home-delivered neonates and their care-givers with the correct knowledge and health-care practices. PMID- 15791270 TI - Requirements of peptidoglycan structure that allow detection by the Drosophila Toll pathway. AB - The Drosophila immune system is able to discriminate between classes of bacteria. Detection of Gram-positive bacteria involves a complex of two pattern recognition receptors: peptidoglycan recognition protein SA (PGRP-SA) and Gram-negative binding protein 1 (GNBP1). These activate the Toll signalling pathway. To define the cell wall components sensed by the host, we used highly purified peptidoglycan fragments of two principal Gram-positive bacterial pathogens Staphylococcus aureus and Streptococcus pneumoniae. We report that in both peptidoglycans, the minimal structure needed to activate the Toll pathway is a muropeptide dimer and that the free reducing end of the N-acetyl muramic acid residues of the muropeptides is essential for activity. Monomeric muropeptides were inactive and inhibitory in combination with dimers. Finally, peptidoglycan was degraded by the haemolymph of wild-type but not GNBP1 mutant flies. We suggest a model whereby GNBP1 is involved in the hydrolysis of Gram-positive peptidoglycan producing new glycan reducing ends, which are subsequently detected by PGRP-SA. PMID- 15791274 TI - The incidence of morbidities in a cohort of neonates in rural Gadchiroli, India: seasonal and temporal variation and a hypothesis about prevention. AB - BACKGROUND: The incidence of morbidities among home-cared neonates in rural areas has not been studied. OBJECTIVES: To estimate the incidence of various neonatal morbidities and the associated risk of death in home-cared neonates in rural setting. To estimate the variation in the incidence of neonatal morbidities by season and by day of life. To identify the scope for prevention of morbidities and suggest a hypothesis. STUDY DESIGN: A prospective observational study nested in the first year of the field trial in rural Gadchiroli, India. Trained village health workers in 39 villages observed neonates at the time of birth and in subsequent eight home visits up to 28 days. We diagnosed 20 neonatal morbidities by using clinical definitions. The data were analyzed for the incidence, case fatality, and relative risk of death and for the seasonal and day-wise variation in the incidence of morbidities. RESULTS: We observed total 763 neonates in 1 year. The incidence of morbidities was a mean of 2.2 morbidities per neonate. The case fatality in 13 morbidities was >10%. Only 2.6% neonates were seen or treated by a physician, and 0.4% were hospitalized. Hypothermia, fever, upper respiratory symptoms, umbilical and skin infections, and conjunctivitis showed statistically significant seasonal variation. Although the morbidities were concentrated in the first week of life, new cases continued to appear throughout the neonatal period. Various morbidities showed different distribution of incidence during 1 to 28 days. CONCLUSIONS: A large burden of disease occurs in rural home-cared neonates, and many morbidities are associated with high case fatality. Some morbidities show strong seasonal and day-wise variation in incidence, indicating poor care at home. We hypothesize that changes in practices and better home-based care will prevent the seasonal and temporal increase in morbidities. Some morbidities may not be preventable and will need early detection and treatment. Therefore, frequent home visits by a health worker are necessary to identify sick neonates. PMID- 15791275 TI - Why do neonates die in rural Gadchiroli, India? (Part I): primary causes of death assigned by neonatologist based on prospectively observed records. AB - OBJECTIVE: To determine the primary causes of death in home-cared rural neonates by using prospectively kept health records of neonates and a neonatologist's clinical judgment. STUDY DESIGN: In the first year (1995 to 1996) of the field trial in Gadchiroli, India, trained village health workers observed neonates in 39 villages by attending home deliveries and making eight home visits during days 0 to 28. The recorded data were validated in the field by a physician. An independent neonatologist assigned the most probable single primary cause of death based on these recorded data. FINDINGS: A total of 763 neonates were observed, of whom 40 died (NMR 52.4/1000). The primary causes of death were sepsis/pneumonia 21 (52.5%), asphyxia 8 (20%), prematurity <32 weeks 6 (15%), hypothermia 1 (2.5%), and other/not known 4 (10%). Most of the prematurity or asphyxia deaths occurred during the first 3 days of life. All 21 sepsis/pneumonia deaths occurred during days 4 to 28. A similar picture existed in England before the antibiotic era. CONCLUSION: Sepsis/pneumonia is the primary cause in half the deaths in rural neonates cared for at home in Gadchiroli, followed by asphyxia and prematurity. Infections cause a larger proportion of deaths in neonates in the community compared to the reported proportion in hospital-based studies. PMID- 15791276 TI - Background of the field trial of home-based neonatal care in Gadchiroli, India. AB - The field trial of home-based neonatal care was conducted in Gadchiroli, India during 1993 to 1998. Owing to its new approach and the success in reducing newborn mortality in a rural area, it has attracted considerable attention. In this article, we describe the background of the trial -- the situation in 1990, why the problems of neonatal mortality and neonatal infection were selected for research, the area -- Gadchiroli district -- where the study was conducted, and the background work and philosophy of the organization, SEARCH, which conducted the study. This history and background will help readers understand the origins and the context of the field trial and the subsequent research papers in this supplement. We also hope that sharing this will be of use to other researchers and program managers working with communities in developing countries. PMID- 15791277 TI - Why do neonates die in rural Gadchiroli, India? (Part II): estimating population attributable risks and contribution of multiple morbidities for identifying a strategy to prevent deaths. AB - OBJECTIVES: The understanding about why neonates die in rural areas in developing countries is limited. In the first year (1995 to 1996) of the field trial of home based neonatal care in rural Gadchiroli, India, we prospectively observed a cohort of neonates in 39 villages. In Part I of this article, we presented the primary causes of death. The data were further analyzed: To estimate the population attributable risk (PAR) of death for the main causes of neonatal mortality. To evaluate the effect of a multiplicity of morbidities and to identify which morbidity combinations cause neonatal deaths. To develop a hypothesis about how best to reduce neonatal mortality. STUDY DESIGN: We analyzed the observational data by logistic regression to estimate the PAR of death for six major morbidities. The effect of the number of morbidities per neonate on case fatality (CF) was estimated. Then we identified the main combinations of morbidities as the component causes leading to death. We estimated the excess deaths attributable to sepsis. RESULTS: This cohort included 763 neonates among whom 40 neonatal deaths occurred. Six major morbidities were associated with the following proportion of deaths: preterm, 62.5%; sepsis, 60%; intrauterine growth restriction (IUGR), 27.5%; asphyxia, 25%; hypothermia, 22.5%, and feeding problems, 15%. The estimated PARs were: preterm, 0.74; IUGR, 0.55; sepsis, 0.55; asphyxia, 0.35; hypothermia, 0.08, and feeding problems, 0.04. The CF associated with the number of morbidities per neonate was: with no morbidity, 0.3%; one morbidity, 2.1%; two morbidities, 15.3%; three or more morbidities, 41.4% (p<0.001). In all, 82.5% of all deaths occurred in neonates with two or more morbidities. The proportion of total deaths associated with only preterm was 7.5%, and with only IUGR was 2.5%; however, with the main morbidity combinations it was preterm+sepsis, 35%; IUGR+sepsis, 22.5%; preterm+asphyxia, 20%; preterm+hypothermia, 15%; and preterm+feeding problem, 12.5%. The % CF with low birth weight (LBW) <2500 g alone was 5.2% and with infection alone was 1.9%, but with LBW+infection it was 31.9%. The estimated excess deaths caused by sepsis over and above LBW was 44% of the total deaths. CONCLUSIONS: Preterm and IUGR are ubiquitous components, but usually not sufficient to cause death. Most deaths occur due to a combination of preterm or IUGR with other comorbidities. If preterm birth or IUGR cannot be prevented, the strategy should be to ensure neonatal survival by addressing comorbidities, that is, infections, asphyxia, hypothermia, and feeding problems in that order of priority. We hypothesize that the prevention and/or management of neonatal infections will reduce neonatal mortality by 40 to 50%. PMID- 15791278 TI - How to identify neonates at risk of death in rural India: clinical criteria for the risk approach. AB - OBJECTIVE: Majority of neonates in developing countries are born at home and most neonatal deaths occur without receiving medical care. This retrospective analysis was undertaken to develop simple clinical criteria for use in rural community to identify neonates at risk of death. STUDY DESIGN: By analyzing the observational data on two cohorts of neonates in 39 villages in different years of the Gadchiroli field trial, we selected a minimum set of clinical features. We evaluated this set for its sensitivity, specificity and predictive value to detect eventual neonatal death, the primary study outcome. RESULTS: The cohorts included 763 neonates with 40 deaths in 1995 to 1996, a year with minimum interventions, and 1598 neonates with 38 deaths in 1996 to 1998, the years of intensive interventions. On the day of birth, presence of any one of the three: (1) birth weight <2000 g, (2) preterm birth or (3) baby not taking feeds; or, during the rest of neonatal life, mother's report of reduced or stopped sucking by baby, were identified as the predictors of neonatal deaths. The combined set gave a sensitivity of 95%, specificity, 77.3%; predictive value, 18.8%; and the yield, 26.5% in 1995 to 1996 and, respectively, 86.8, 78, 8.8, and 23.5% in 1996 to 1998. The mean lead time gained was 3.4 to 6.6 days. CONCLUSION: Presence of any one of the four predictors will identify with high sensitivity and moderate specificity nearly a quarter of the neonates in rural community as high risk, 3.4 to 6.6 days in advance, for intensive attention at home or referral. PMID- 15791279 TI - Reduced incidence of neonatal morbidities: effect of home-based neonatal care in rural Gadchiroli, India. AB - OBJECTIVE: We found a high burden of morbidities in a cohort of neonates observed in rural Gadchiroli, India. We hypothesised that interventions would reduce the incidence of neonatal morbidities, including the seasonal increase observed in many of them. This article reports the effect of home-based neonatal care on neonatal morbidities in the intervention arm of the field trial by comparing the early vs late periods, and the possible explanation for this effect. METHODS: During 3 years (1995 to 1998), trained village-health-workers (VHWs) in 39 villages prospectively collected data by making home visits during pregnancy, home-delivery and during neonatal period. We estimated the incidence and burden of neonatal morbidities over the 3 years from these data. In the first year, the VHWs made home visits only to observe. From the second year, they assisted mothers in neonatal care and managed the sick neonates at home. Health education of mothers and family members, individually and in group, was added in the third year. We measured the coverage of interventions over the 3 years and evaluated maternal knowledge and practices on 21 indicators in the third year. The effect on 17 morbidities was estimated by comparing the incidence in the first year with the third year. RESULTS: The VHWs observed 763 neonates in the first year, 685 in the second and 913 in the third year. The change in the percent incidence of morbidities was (i) infections, from 61.6 to 27.5 (-55%; p<0.001), (ii) care related morbidities (asphyxia, hypothermia, feeding problems) from 48.2 to 26.3 ( 45%; p<0.001); (iii) low birth weight from 41.9 to 35.2 (-16%; p<0.05); (iv) preterm birth and congenital anomalies remained unchanged. The mean number of morbidities/100 neonates in the 3 years was 228, 170 and 115 (a reduction of 49.6%; p<0.001). These reductions accompanied an increasing percent score of interventions during 3 years: 37.9, 58.4 and 81.3, thus showing a dose-response relationship. In the third year, the proportion of correct maternal knowledge was 78.7% and behaviours was 69.7%. The significant seasonal increase earlier observed in the incidence of five morbidities reduced in the third year. CONCLUSION: The home-based care and health education reduced the incidence and burden of neonatal morbidities by nearly half. The effect was broad, but was especially pronounced on infections, care-related morbidities and on the seasonal increase in morbidities. PMID- 15791280 TI - Is home-based diagnosis and treatment of neonatal sepsis feasible and effective? Seven years of intervention in the Gadchiroli field trial (1996 to 2003). AB - OBJECTIVES: To further evaluate our earlier findings on the feasibility and effectiveness of home-based management of neonatal sepsis by analysing 7 years data (1996 to 2003) from the field trial in Gadchiroli, India. STUDY DESIGN: Neonates in 39 villages were monitored by trained village health workers (VHWs) from 1995 onwards. In 1996, we trained VHWs to diagnose sepsis by using a clinical algorithm and provide domiciliary treatment using intramuscular gentamicin and oral co-trimoxazole. Health records for all neonates were kept by the VHWs, checked by field supervisors, and computerized. Live births and neonatal deaths were recorded by an independent vital statistics collection system. We evaluated the feasibility and effectiveness of this approach. RESULTS: During September 1996 to March 2003, VHWs monitored 93% of all neonates in 39 villages (N=5268). As compared to 552 cases of sepsis diagnosed by computer algorithm, VHWs correctly diagnosed 492 cases (89%). Parents agreed to home-based treatment for the majority of infants (448, 91%), refused treatment in 31 (6.4%) cases, and hospitalized 13 infants (2.6%). VHWs treated 470 neonates with antibiotics, that is, 8.9% of all neonates in community. Of 552 cases diagnosed by computer, VHWs correctly treated 448 (81.2%) and gave unnecessary treatment to 22/470 (4.7%) of treated neonates. The case fatality (CF) was 6.9% in treated cases vs 22% in untreated or 16.6% in the pre-intervention period (p<0.001). Home based treatment resulted in 67.2% reduction in %CF among preterm and a 72% reduction among LBW neonates. CONCLUSIONS: Home-based management of neonates with suspected sepsis is acceptable to most parents, safe, and effective in reducing sepsis case fatality by nearly 60%. With proper selection, training, and supervision of health workers, this method may be applicable in areas in developing countries where access to hospital care is limited. PMID- 15791281 TI - Low birth weight and preterm neonates: can they be managed at home by mother and a trained village health worker? AB - OBJECTIVE: Observations on a cohort of neonates in the preintervention year of the field trial of home-based neonatal care (HBNC) in rural Gadchiroli, India, showed that preterm birth and low birth weight (LBW), <2500 g, constituted the most important risk factors. Owing to a limited access to hospital care, most neonates were managed at home in the subsequent intervention years. The objective of this paper is to evaluate the feasibility and effectiveness of managing LBW and preterm neonates in home setting. DESIGN: We retrospectively analyzed data from the intervention arm (39 villages) in the HBNC trial. Feasibility was assessed by coverage and by quality (19 indicators) of care. Effectiveness was evaluated by change in case fatality (CF) and in the incidence of comorbidities in LBW or preterm neonates by comparing the preintervention year (1995 to 1996) with the intervention years (1996 to 2003). RESULTS: During 1996 to 2003, total 5919 live births occurred in the intervention villages, out of whom 5510 (93%) received HBNC. These included 2015 LBW neonates and 533 preterm neonates, out of whom 97% received only home-based care. The coverage and quality of interventions assessed on 19 indicators was 80.5%. The CF in LBW neonates declined by 58% (from 11.3 to 4.7%, p<0.001), and in preterm neonates, by 69.5% (from 33.3 to 10.2%, p<0.0001). Incidence of the major comorbidities, viz., sepsis, asphyxia, hypothermia and feeding problems, declined significantly. Preterm-LBW neonates without sepsis (270) received only supportive care -- CF in them decreased from 28.2 to 11.5% (p<0.01), and those with sepsis (53) received supportive care and antibiotics -- CF in them decreased from 61 to 13.2% (p<0.005). Supportive care contributed 75% and treatment with antibiotics 25% in the total averted deaths in preterm-LBW neonates. The intrauterine growth restriction (IUGR)-LBW neonates without sepsis (1409) received only supportive care -- the CF was unchanged, and 181 with sepsis received supportive care and antibiotics -- the CF decreased from 18.4 to 8.8% (p<0.05). Treatment with antibiotics explained entire reduction in mortality in IUGR neonates. In total, 55 deaths in LBW neonates were averted by supportive care and 35 by the treatment with antibiotics. CONCLUSIONS: Home-based management of LBW and the preterm neonates is feasible and effective. It remarkably improved survival by preventing comorbidities, by supportive care, and by treating infections. PMID- 15791282 TI - Management of birth asphyxia in home deliveries in rural Gadchiroli: the effect of two types of birth attendants and of resuscitating with mouth-to-mouth, tube mask or bag-mask. AB - OBJECTIVES: To evaluate the effect of home-based neonatal care on birth asphyxia and to compare the effectiveness of two types of workers and three methods of resuscitation in home delivery. STUDY DESIGN: In a field trial of home-based neonatal care in rural Gadchiroli, India, birth asphyxia in home deliveries was managed differently during different phases. Trained traditional birth attendants (TBA) used mouth-to-mouth resuscitation in the baseline years (1993 to 1995). Additional village health workers (VHWs) only observed in 1995 to 1996. In the intervention years (1996 to 2003), they used tube-mask (1996 to 1999) and bag mask (1999 to 2003). The incidence, case fatality (CF) and asphyxia-specific mortality rate (ASMR) during different phases were compared. RESULTS: During the intervention years, 5033 home deliveries occurred. VHWs were present during 84% home deliveries. The incidence of mild birth asphyxia decreased by 60%, from 14% in the observation year (1995 to 1996) to 6% in the intervention years (p<0.0001). The incidence of severe asphyxia did not change significantly, but the CF in neonates with severe asphyxia decreased by 47.5%, from 39 to 20% (p<0.07) and ASMR by 65%, from 11 to 4% (p<0.02). Mouth-to-mouth resuscitation reduced the ASMR by 12%, tube-mask further reduced the CF by 27% and the ASMR by 67%. The bag-mask showed an additional decrease in CF of 39% and in the fresh stillbirth rate of 33% in comparison to tube-mask (not significant). The cost of bag and mask was US dollars 13 per averted death. Oxytocic injection administered by unqualified doctors showed an odds ratio of three for the occurrence of severe asphyxia or fresh stillbirth. CONCLUSIONS: Home-based interventions delivered by a team of TBA and a semiskilled VHW reduced the asphyxia-related neonatal mortality by 65% compared to only TBA. The bag-mask appears to be superior to tube-mask or mouth-to-mouth resuscitation, with an estimated equipment cost of US dollars 13 per death averted. PMID- 15791283 TI - Neonatal and infant mortality in the ten years (1993 to 2003) of the Gadchiroli field trial: effect of home-based neonatal care. AB - OBJECTIVES: To evaluate the effect on neonatal and infant mortality during 10 years (1993 to 2003) in the field trial of home-based neonatal care (HBNC) in Gadchiroli. To estimate the contribution of the individual components in the intervention package on the observed effect. STUDY DESIGN: The field trial of HBNC in Gadchiroli, India, has completed the baseline phase (1993 to 1995), observational phase (1995 to 1996) and the 7 years of intervention (1996 to 2003). We measured the stillbirth rate (SBR), neonatal mortality rate (NMR), perinatal mortality rate (PMR), postneonatal mortality rate (PNMR) and the infant mortality rate (IMR) in the intervention area and the control area. The effect of HBNC on all these rates was estimated by comparing the change from baseline (1993 to 1995) to the last 2 years of intervention (2001 to 2003) in the intervention area vs in the control area. For other estimates, we made a before-after comparison of the rates in the intervention arm in the observation year (1995 to 1996) vs intervention years (1996 to 2003). We evaluated the effect on the cause specific NMRs. By using the changes in the incidence and case fatality (CF) of the four main morbidities, we estimated the contribution of primary prevention and of the management of sick neonates. The proportion of deaths averted by different components of HBNC was estimated. RESULTS: The baseline population in 39 intervention villages was 39,312 and in 47 control villages it was 42,617, and the population characteristics and vital rates were similar. The total number of live births in 10 years (1993 to 2003) were 8811 and 9990, respectively. The NMR in the control area showed an increase from 58 in 1993 to 1995 to 64 in 2001 to 2003. The NMR in the intervention area declined from 62 to 25; the reduction in comparison to the control area was by 44 points (70%, 95% CI 59 to 81%). Early NMR decreased by 24 points (64%) and late NMR by 20 points (80%). The SBR decreased by 16 points (49%) and the PMR by 38 points (56%). The PNMR did not change, and the IMR decreased by 43 points (57%, 95% CI 46 to 68%). All reductions were highly significant (p<0.001) except for SBR it was <0.05. The cause-specific NMR (1995 to 1996 vs 2001 to 2003) for sepsis decreased by 90%, for asphyxia by 53% and for prematurity by 38%. The total reduction in neonatal mortality during intervention (1996 to 2003) was ascribed to sepsis management, 36%; supportive care of low birth weight (LBW) neonates, 34%; asphyxia management, 19%; primary prevention, 7% and management of other illnesses or unexplained, 4%. CONCLUSIONS: The HBNC package in the Gadchiroli field trial reduced the neonatal and perinatal mortality by large margins, and the gains were sustained at the end of the 7 years of intervention and were carried forward as improved survival through the first year of life. Most of the reduction in mortality was ascribed to sickness management, that is, management of sepsis, supportive care of LBW neonates and management of asphyxia, in that order, and a small portion to primary prevention. PMID- 15791284 TI - Gene expression alterations in the non-neoplastic parenchyma adjacent to infiltrating pancreatic ductal adenocarcinoma. AB - The non-neoplastic pancreatic parenchyma adjacent to infiltrating ductal adenocarcinoma demonstrates inflammation, fibrosis, acinar cell loss and small duct-like metaplasia of acinar cells. Similar morphologic changes are also observed in the setting of chronic pancreatitis. In addition, peritumoral acini have been shown to have alterations in gene expression even in the absence of morphological changes. To better understand the pancreatic acinar responses to infiltrating pancreatic ductal adenocarcinoma, we characterized gene expression patterns of pancreatic acinar tissue adjacent to infiltrating pancreatic ductal adenocarcinomas and compared them to gene expression patterns of acinar tissue affected by chronic pancreatitis as well as to those of normal pancreatic acini. Fresh-frozen pancreatic acinar tissue was microdissected from nine patients (three with pancreatic cancer, three with chronic pancreatitis, three with normal pancreata) using laser capture microdissection, and extracted RNA from each microdissection was subjected to two rounds of linear amplification and hybridized to oligonucleotide microarrays. Gene expression patterns were confirmed using quantitative RT-PCR and/or immunohistochemistry. A total of 20 genes was found to be overexpressed in peritumoral acinar tissue compared to normal acinar tissue and to acini affected by chronic pancreatitis. These 20 genes included pancreatitis-associated protein (HIP/PAP), a gene known to be overexpressed in acini adjacent to infiltrating pancreatic cancer, and the gene cartilage glycoprotein-39 (HC gp-39 or TKL-40). Serum HC gp-39 protein levels were significantly higher in patients with pancreatic cancer and in those with chronic pancreatitis than in controls without pancreatic disease. There was no significant difference in the levels of serum HC gp-39 in patients with pancreatic cancer and those with chronic pancreatitis. Our results demonstrate some of the molecular alterations in acinar cells that occur in response to adjacent infiltrating pancreatic ductal adenocarcinoma and reveal that such alterations can provide a rich source of markers of pancreatic cancer. PMID- 15791285 TI - Cardiac chymase: pathophysiological role and therapeutic potential of chymase inhibitors. AB - On release from cardiac mast cells, alpha-chymase converts angiotensin I (Ang I) to Ang II. In addition to Ang II formation, alpha-chymase is capable of activating TGF-beta1 and IL-1beta, forming endothelins consisting of 31 amino acids, degrading endothelin-1, altering lipid metabolism, and degrading the extracellular matrix. Under physiological conditions the role of chymase in the mast cells of the heart is uncertain. In pathological situations, chymase may be secreted and have important effects on the heart. Thus, in animal models of cardiomyopathy, pressure overload, and myocardial infarction, there are increases in both chymase mRNA levels and chymase activity in the heart. In human diseased heart homogenates, alterations in chymase activity have also been reported. These findings have raised the possibility that inhibition of chymase may have a role in the therapy of cardiac disease. The selective chymase inhibitors developed to date include TY-51076, SUN-C8257, BCEAB, NK320, and TEI-E548. These have yet to be tested in humans, but promising results have been obtained in animal models of myocardial infarction, cardiomyopathy, and tachycardia-induced heart failure. It seems likely that orally active inhibitors of chymase could have a place in the treatment of cardiac diseases where injury-induced mast cell degranulation contributes to the pathology. PMID- 15791286 TI - Examining the interaction of apo E and neurotoxicity on a murine model of ALS PDC. AB - Epidemiological studies have shown a positive relationship between cycad flour consumption and the development of the neurodegenerative disorder, amyotrophic lateral sclerosis - parkinsonism - dementia complex (ALS-PDC). Apolipoprotein E (apo E) allele variations have been associated with genetic susceptibility in neurodegenerative diseases, including ALS-PDC. We have studied cycad toxicity in a mouse model of ALS-PDC with a particular interest in its impact on the central nervous system (CNS) in both apo E knock-out (KO) mice and their wild-type (WT) counterparts. Behavioral motor tests, motor neuron counts, and immunohistochemical staining in brain and spinal cord, as well as routine histological examinations on internal organs, were performed to evaluate cycad toxicity. Plasma cholesterol levels were also measured before and during the study. Cycad treatment was associated with higher levels of plasma cholesterol only in apo E KO mice; increased levels of plasma cholesterol did not result in increased athero genesis. Cycad-fed wild-type mice developed progressive behavioral deficits including ALS-PDC-like pathological outcomes, while cycad-fed apo E KO mice were not significantly affected. Cycad-fed wild-type mice had shorter gait length measurements along with higher active caspase-3 levels in the striatum, substantia nigra, primary motor cortex, and spinal cord as compared with corresponding controls. These changes were associated with decreased labeling for glutamate transporter 1B and tyrosine hydroxylase activity levels. No evidence of cycad toxicity was observed in internal organs of either wild-type or apo E KO mice. Our data demonstrate that apo E KO mice are less susceptible to cycad toxicity, suggesting a role for apo E as a possible genetic susceptibility factor for some forms of toxin-induced neurodegeneration. PMID- 15791287 TI - Co-expression and regulation of connexins 36 and 43 in cultured neonatal rat pancreatic islets. AB - Fetal and neonatal pancreatic islets present a lower insulin secretory response as compared with adult islets. Prolonged culturing leads to an improvement of the glucose-induced insulin secretion response in neonatal pancreatic islets that may involve regulation of gap junction mediated cell communication. In this study, we investigated the effect of culturing neonatal islet cells for varying periods of time and with different glucose medium concentrations on the cellular expression of the endocrine pancreatic gap junction associated connexin (Cx) 36 and Cx43. We report here that the 7-d culture induced upregulation of the expression of these junctional proteins in neonatal islets in a time-dependent manner. A correlation was observed between the increased mRNA and protein expression of Cx36 and Cx43 and the increased insulin secretion following islet culturing. In addition, increasing glucose concentration within the culture medium induced a concentration-dependent enhancement of Cx36 islet expression, but not of Cx43 expression in cultured neonatal islets. In conclusion, we suggest that the regulation of gap junctional proteins by culture medium containing factors and glucose may be an important event for the maturation process of beta cells observed at in vitro conditions. PMID- 15791288 TI - The Gurvich waveform has lower defibrillation threshold than the rectilinear waveform and the truncated exponential waveform in the rabbit heart. AB - Implantable cardioverter defibrillator studies have established the superiority of biphasic waveforms over monophasic waveforms. However, external defibrillator studies of biphasic waveforms are not as widespread. Our objective was to compare the defibrillation efficacy of clinically used biphasic waveforms, i.e., truncated exponential, rectilinear, and quasi-sinusoidal (Gurvich) waveforms in a fibrillating heart model. Langendorff-perfused rabbit hearts (n = 10) were stained with a voltage-sensitive fluorescent dye, Di-4-ANEPPS. Transmembrane action potentials were optically mapped from the anterior epicardium. We found that the Gurvich waveform was significantly superior (p < 0.05) to the rectilinear and truncated exponential waveforms. The defibrillation thresholds (mean +/- SE) were as follows: Gurvich, 0.25 +/- 0.01 J; rectilinear-1, 0.34 +/- 0.01 J; rectilinear-2, 0.33 +/- 0.01 J; and truncated exponential, 0.32 +/- 0.02 J. Using optically recorded transmembrane responses, we determined the shock response transfer function, which allowed us to predict the cellular response to waveforms at high accuracy. The passive parallel resistor-capacitor model (RC model) predicted polarization superiority of the Gurvich waveform in the myocardium with a membrane time constant (taum) of less than 2 ms. The finding of a lower defibrillation threshold with the Gurvich waveform in an in vitro model of external defibrillation suggests that the Gurvich waveform may be important for future external defibrillator designs. PMID- 15791289 TI - Reductive effect of body weight in C57BL/6J mice fed a high-fat diet by Saesaengmyung Diet. AB - The Saesaengmyung Diet (SD) is a newly developed dietary product to help control weight. The aim of this study was to evaluate whether SD combined with a high-fat (HF) diet could influence body weight, fat accumulation, and glucose levels in blood. C57BL/6J mice were fed for 8 weeks with a standard diet, an HF diet, and an HF + 10% or HF + 20% SD diet. Body weight was recorded weekly, and plasma levels of total cholesterol, low-density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol, and glucose were analyzed at the end of the study. Weight increases in the 10% or 20% SD group were significantly less than in the HF diet group (p < 0.05). Plasma total cholesterol level significantly decreased by 33.5% in the 10% SD group and 38.8% in the 20% SD group, but the LDL cholesterol, HDL cholesterol, and glucose levels in the SD groups were not significantly changed. Our findings indicate that SD may be beneficial to overweight individuals in the reduction of weight gain induced by an HF diet. PMID- 15791290 TI - The cardioprotective effect of dual metallopeptidase inhibition: respective roles of endogenous kinins and natriuretic peptides. AB - The objective of the present study was to assess the cardioprotective effect of dual NEP-ACE inhibition in relation to endogenous cardiac bradykinin (BK), its active metabolite des-Arg9-BK, endogenous brain natriuretic peptides (BNP), and cGMP. Rats were treated with the dual metallopeptidase inhibitor, omapatrilat, or the ACE inhibitor, ramipril, for 7 d (1 mg.kg(-1).d(-1)). Hearts were then isolated and subjected to a zero-flow ischemia and reperfusion (except controls), in the absence or presence of either a B2-receptor antagonist (Hoe-140), a B1 receptor antagonist (Lys-Leu8-des-Arg9-BK), or the GC-A/GC-B-receptor antagonist (HS-142-1). Chronic omapatrilat and ramipril increased the amount of endogenous BK collected upon reperfusion, but only ramipril increased that of des-Arg9-BK. Only omapatrilat increased both peak BNP and peak cGMP upon reperfusion, those increases being blocked by Hoe-140. Chronic omapatrilat (but not ramipril) decreased the total noradrenaline and lactate dehydrogenase release during the reperfusion period. Importantly, only omapatrilat improved the functional recovery of the ischemic reperfused heart, with a reduced left ventricular end diastolic pressure, and improved developed left ventricular pressure. All cardio protective effects of omapatrilat were blocked by Hoe-140 and by HS-142-1, but not by the B1-receptor antagonist. In conclusion, a chronic treatment with a dual metallopeptidase inhibitor demonstrated a cardioprotective action not observed with an ACE inhibitor in a context of severe ischemia in rat isolated hearts, which was mediated by both endogenous BK and BNP. PMID- 15791291 TI - The low oxygen-carrying capacity of Krebs buffer causes a doubling in ventricular wall thickness in the isolated heart. AB - The buffer-perfused Langendorff heart is significantly vasodilated compared with the in vivo heart. In this study, we employed ultrasound to determine if this vasodilation translated into changes in left ventricular wall thickness (LVWT), and if this effect persisted when these hearts were switched to the "working" mode. To investigate the effects of perfusion pressure, vascular tone, and oxygen availability on cardiac dimensions, we perfused hearts (from male Wistar rats) in the Langendorff mode at 80, 60, and 40 cm H2O pressure, and infused further groups of hearts with either the vasoconstrictor endothelin-1 (ET-1) or the blood substitute FC-43. Buffer perfusion induced a doubling in diastolic LVWT compared with the same hearts in vivo (5.4 +/- 0.2 mm vs. 2.6 +/- 0.2 mm, p < 0.05) that was not reversed by switching hearts to "working" mode. Perfusion pressures of 60 and 40 cm H2O resulted in an increase in diastolic LVWT. ET-1 infusion caused a dose-dependent decrease in diastolic LVWT (6.6 +/- 0.4 to 4.8 +/- 0.4 mm at a concentration of 10(-9) mol/L, p < 0.05), with a concurrent decrease in coronary flow. FC-43 decreased diastolic LVWT from 6.7 +/- 0.5 to 3.8 +/- 0.7 mm (p < 0.05), with coronary flow falling from 16.1 +/- 0.4 to 8.1 +/- 0.4 mL/min (p < 0.05). We conclude that the increased diastolic LVWT observed in buffer-perfused hearts is due to vasodilation induced by the low oxygen-carrying capacity of buffer compared with blood in vivo, and that the inotropic effect of ET-1 in the Langendorff heart may be the result of a reversal of this wall thickening. The implications of these findings are discussed. PMID- 15791292 TI - Effect of BM 17.0744, a PPARalpha ligand, on the metabolism of perfused hearts from control and diabetic mice. AB - Peroxisome proliferator-activated receptor-alpha (PPARalpha) regulates the expression of fatty acid (FA) oxidation genes in liver and heart. Although PPARalpha ligands increased FA oxidation in cultured cardiomyocytes, the cardiac effects of chronic PPARalpha ligand administration in vivo have not been studied. Diabetic db/db mouse hearts exhibit characteristics of a diabetic cardiomyopathy, with altered metabolism and reduced contractile function. A testable hypothesis is that chronic administration of a PPARalpha agonist to db/db mice will normalize cardiac metabolism and improve contractile function. Therefore, a PPARalpha ligand (BM 17.0744) was administered orally to control and type 2 diabetic (db/db) mice (37.9 +/- 2.5 mg/(kg.d) for 8 weeks), and effects on cardiac metabolism and contractile function were assessed. BM 17.0744 reduced plasma glucose in db/db mice, but no change was observed in control mice. FA oxidation was significantly reduced in BM 17.0744 treated db/db hearts with a corresponding increase in glycolysis and glucose oxidation; glucose and FA oxidation in control hearts was unchanged by BM 17.0744. PPARalpha treatment did not alter expression of PPARalpha target genes in either control or diabetic hearts. Therefore, metabolic alterations in hearts from PPARalpha-treated diabetic mice most likely reflect indirect mechanisms related to improvement in diabetic status in vivo. Despite normalization of cardiac metabolism, PPARalpha treatment did not improve cardiac function in diabetic hearts. PMID- 15791293 TI - Repeated daily injections and osmotic pump infusion of isoproterenol cause similar increases in cardiac mass but have different effects on blood pressure. AB - We found in mice that repeated single daily subcutaneous (s.c.) isoproterenol (ISO) injections, like constant infusions using osmotic minipumps, caused increased biventricular mass or weight relative to body weight (VW/BW). We found that 5 (1/d) s.c. injections of 2, 10, or 20 microg/g body weight caused equivalent VW/BW increases as compared with 5-d infusions at 20 microg/(g.d)). While it is often presumed that ISO elicits hypertrophy by a direct effect on the myocytes, growth may also be secondary to systemic hemodynamic effects. The 2 modes of ISO administration had different effects on mean arterial blood pressure (MABP) and heart rate. Using telemetry we observed that single injections of ISO (0, 0.5, 2, and 10 microg/g) were associated with hypotension and tachycardia with a duration but not a magnitude that was dose dependent. MABP dropped rapidly to 60 mm Hg for more than 2 h at the highest dose. Constant s.c. infusion of ISO at 20 microg/(g.d) initially lowered MABP to about 70 mm Hg for 24 h. At 48 h MABP was normal, but rose 10 mm Hg higher than baseline by day 5. Thus, different routes of administration of ISO that cause comparable increases in VW/BW had different effects on MABP. Thus when evaluating mouse models of ISO-induced cardiac hypertrophy, both repeated daily injections or infusions can cause similar increases in VW/BW, but the daily doses that are required are not the same. Furthermore, these different routes of administration have different hemodynamic sequelae and could potentially evoke different cardiac phenotypes. PMID- 15791295 TI - Carborane acids. New "strong yet gentle" acids for organic and inorganic chemistry. AB - Icosahedral carborane anions such as CHB11Cl11- are amongst the least coordinating, most chemically inert anions known. They are also amongst the least basic, so their conjugate acids, H(carborane), are superacids (i.e. stronger than 100% H2SO4). Acidity scale measurements indicate that H(CHB11Cl11) is the strongest pure Bronsted acid presently known, surpassing triflic and fluorosulfuric acid. Nevertheless, it is also an extremely gentle acid--because its conjugate base engages in so little chemistry. Carborane acids separate protic acidity from anion nucleophilicity and destructive oxidative capacity in the conjugate base, to a degree not previously achieved. As a result, many long sought, highly acidic, reactive cations such as protonated benzene (C6H7+), protonated C60(HC60+), tertiary carbocations (R3C+), vinyl cations (R2C=C(+)-R), silylium ions (R3Si+) and discrete hydronium ions (H3O+, H5O2+ etc.) can be readily isolated as carborane salts and characterized at room temperature by X ray crystallography. PMID- 15791294 TI - Primary infection of mice with high titer inoculum respiratory syncytial virus: characterization and response to antiviral therapy. AB - Intranasal infection of BALB/c mice with respiratory syncytial virus (RSV)-A2 (0.5 x 10(8) - 2.0 x 10(8) plaque-forming units, PFU) produced disease characterized by weight loss (2-3 g) and mortality (60%-100%) with the mean day of death ranging from 6-7 d after infection. The extent of RSV disease was inoculum titer-dependent and required a replication competent virus. Lung titers of virus peaked at 0.5-1 x 10(6) PFU/g wet weight. Bronchoalveolar lavage fluid (BALF) levels of IL-1beta, TNF-alpha, INF-gamma IL-12, IL-6, MIP-1alpha, RANTES, and protein were elevated, whereas IL-2, IL-4, IL-5, IL-13, and IL-10 were unchanged. Histological assessment of lungs revealed marked inflammatory pathology characterized by bronchiolitis, vasculitis, and interstitial pneumonia. Whole-body plethysmography revealed significant disease-associated deficits of respiratory function. Therapy with ribavirin administered either by the intranasal, subcutaneous, or oral route significantly reduced disease in a dose dependent manner. Delaying the initiation of therapy resulted in a loss of activity for ribavirin. Synagis administered either intramuscularly as a single dose in prophylaxis or intranasally in prophylaxis, followed by therapy, also significantly reduced disease in a dose-dependent manner. Infection of mice with a high titer inoculum of RSV-A2 resulted in severe and fatal pulmonary disease that was responsive to treatment. This model may be useful to characterize the in vivo activity of experimental therapies for RSV infection. PMID- 15791296 TI - Complex morphology in a simple chemical system. AB - Chemical systems, far from thermodynamic equilibrium, may spontaneously self construct complex structures mimicking biological structures. PMID- 15791297 TI - Manipulating 2D metal-organic networks via ligand control. AB - High-resolution scanning tunneling microscopy has revealed how ligand control can be successfully employed to eliminate isomeric phases and defects in 2D coordination networks that are self-assembled at a surface support by replacing symmetric dicarboxylato linker ligands to dissymmetric carboxylpyridyl linker ligands. PMID- 15791298 TI - Hollow gold and platinum nanoparticles by a transmetallation reaction in an organic solution. AB - Transmetallation reaction between hydrophobized silver nanoparticles with hydrophobized chloroaurate and chloroplatinate ions in chloroform results in the formation of hollow gold and platinum shell nanoparticles respectively. PMID- 15791299 TI - Biomimetic synthesis of (+/-)-9,10-deoxytridachione. AB - A tandem Suzuki-coupling/electrocyclisation reaction sequence was employed for the biomimetic synthesis of (+/-)-9,10-deoxytridachione. PMID- 15791300 TI - A fluorescent pyrophosphate sensor via excimer formation in water. AB - A new fluorescent sensor based on a pyrene/Zn(II)-dpa (dpa = bis(2 pyridylmethyl)amine) conjugate displays excimer emission selective for pyrophosphate over other anions. PMID- 15791301 TI - Remarkably improved complexation of a bisparaquat by formation of a pseudocryptand-based [3]pseudorotaxane. AB - Significant improvement of complexation of a bisparaquat guest was achieved by the formation of a pseudocryptand-based [3]pseudorotaxane. PMID- 15791302 TI - A supramolecular poly[3]pseudorotaxane by self-assembly of a homoditopic cylindrical bis(crown ether) host and a bisparaquat derivative. AB - A supramolecular poly[3]pseudorotaxane was prepared by self-assembly of a homoditopic cylindrical bis(crown ether) host and a bisparaquat derivative in solution by host-guest complexation. PMID- 15791303 TI - Wafer-scale fabrication of periodic polymer attolitre microvial arrays. AB - Wafer-size periodic polymer attolitre microvial arrays of varying depth have been fabricated by templating from spin-coated 2D non-close-packed colloidal crystal polymer nanocomposites. PMID- 15791304 TI - The construction of a supramolecular polymeric rotaxane from bipyridine-ruthenium and cyclodextrin. AB - A luminescent supramolecular link is constructed by a very simple method using bipyridine-ruthenium and cyclodextrin, which displays not only a quasi-linear structure, but also a satisfactory fluorescence emission in both solution and the solid state. PMID- 15791306 TI - Cuprophilic and pi-stacking interactions in the formation of supramolecular stacks from dicoordinate organocopper complexes. AB - The unsupported organocopper pyridine complexes C6F5Cu(py) (2) and [C6F5Cu]2(4,4' bipy) (3) form supramolecular structures that are unprecedented in organocopper chemistry; one-dimensional chains of copper atoms with Cu...Cu distances of 2.8924(3) angstroms in the blue-luminescent complex are likely associated with cuprophilic interactions, whereas multiple perfluoroarene-arene interactions dominate in the supramolecular assembly of 3. PMID- 15791305 TI - Optimized DNA targeting using N,N-bis(2-pyridylmethyl)-beta-alanyl 2'-amino-LNA. AB - Incorporation of N,N-bis(2-pyridylmethyl)-beta-alanyl 2'-amino-LNA (bipyridyl functionalized 2'-amino locked nucleic acid) monomers into DNA strands enables high-affinity targeting of complementary DNA with excellent Watson-Crick selectivity in the presence of divalent metal ions. Positioning of bipyridyl functionalized 2'-amino-LNA monomers in two complementary DNA strands in a "3' end zipper" constitution allows modulation of duplex stability, i.e., a strong stabilizing effect with one equivalent of divalent metal ion per bipyridyl pair, or a strong destabilizing effect with an excess of divalent metal ions. PMID- 15791307 TI - Catalytic enantioselective conjugate addition of dialkylzinc reagents to N substituted-2,3-dehydro-4-piperidones. AB - The first, highly enantioselective, copper/phosphoramidite-catalyzed conjugate addition of dialkylzinc reagents to N-substituted 2,3-dehydro-4-piperidones is described. PMID- 15791308 TI - A cross-linked reverse micelle-encapsulated palladium catalyst. AB - Cross-linked reverse micelle-palladium catalysts are effective and stable cross coupling catalysts; cross-linking is crucial for stability. PMID- 15791309 TI - Nanocoating of natural cellulose fibers with conjugated polymer: hierarchical polypyrrole composite materials. AB - Astonishingly uniform polypyrrole nanocoating on morphologically complex cellulosic substances was achieved without disrupting the hierarchical network structures of individual cellulose fibers by means of polymerization-induced adsorption. PMID- 15791310 TI - Selective recognition of dihydrogen phosphate by receptors bearing pyridyl moieties as hydrogen bond acceptors. AB - Dihydrogen phosphate anion is selectively recognized by amide-based receptors bearing pyridyl moieties as hydrogen bond acceptors in 0.5% DMSO-acetonitrile. PMID- 15791311 TI - Control over the responsive wettability of poly(N-isopropylacrylamide) film in a large extent by introducing an irresponsive molecule. AB - An irresponsive but very hydrophobic molecule--fluoroalkylsilane--can be used to cooperate with PNIPAAm to obtain tunable and enhancedly responsive wettability, and thus largely extend the application scope of PNIPAAm film. PMID- 15791312 TI - Assembling carbon nanotubosomes using an emulsion-inversion technique. AB - Novel micro-capsules (carbon nanotubosomes) have been fabricated by cross-linking shells of amine-functionalized multi-walled carbon nanotubes (MWNTs) produced by their adsorption on water-in-oil emulsion drops followed by an emulsion inversion. PMID- 15791313 TI - Regioselective Si-C bond activation in silicon-bridged ansa-cycloheptatrienyl cyclopentadienyl complexes. AB - On treatment with [Pt(PEt3)3], silicon-bridged ansa-cycloheptatrienyl cyclopentadienyl Ti and V complexes, [1]silatroticenophane and [1]silatrovacenophane, undergo oxidative addition and regioselective insertion of a Pt(PEt3)2 moiety into the silicon-carbon bond to the seven-membered ring; the resulting complexes, [2]platinasilatroticenophane and [2]platinasilatrovacenophane, can be partly used as single-source catalysts for the ring-opening polymerization (ROP) of the original highly strained sandwich molecules. PMID- 15791314 TI - Hydrothermal synthesis of crystalline rectangular titanoniobate particles. AB - Potassium titanoniobate (KTiNbO5) crystalline powders possessing rectangular particle shapes and large surface areas which are prerequisite for high photocatalytic performance have been successfully synthesized by a novel hydrothermal method. PMID- 15791315 TI - Phase-transfer of CdSe@ZnS quantum dots using amphiphilic hyperbranched polyethylenimine. AB - A new, straightforward method for the phase-transfer of CdSe@ZnS quantum dots from non-polar solvents into water and short-chained alcohols using amphiphilic hyperbranched polyethylenimine of different molar weights is suggested and the experimental procedure is discussed as well as the chemical properties of the resulting polymer-derivatised nanocrystals. PMID- 15791316 TI - Improving pulse sequences for 3D DOSY: COSY-IDOSY. AB - An improved pulse sequence for measuring diffusion-ordered COSY spectra is achieved by incorporating the diffusion encoding directly into the evolution and detection periods of a gradient-enhanced COSY sequence, giving improved sensitivity and a 32-fold reduction in minimum experiment time. PMID- 15791317 TI - Controlled synthesis of nanorods/nanorings of a novel Co-Cu complex in microemulsion at room temperature. AB - Novel Co-Cu complex nanorods with diameters of 100-200 nm and nanorings with a ring-diameter of 80 nm were synthesized via a microemulsion method at room temperature. Using this method, the addition of Co(NH3)(3+)6 to aqueous solutions of Cu(ii) in excessive carbonate promotes the assembly of a new highly charged carbonato-copper(ii) anion, [Cu4(OH)(CO3)8]9-. PMID- 15791318 TI - Thermally stable potassium N-heterocyclic carbene complexes with alkoxide ligands, and a polymeric crystal structure with distorted, bridging carbenes. AB - Alkoxide-functionalized N-heterocyclic carbenes (NHCs) stabilise the potassium imidazole-2-ylidene fragment against 1,2 alkyl rearrangement reactions; this allows the first structural characterisation of a potassium NHC complex, which reveals distorted and unusual bonding interactions between the imidazole ring and the potassium cation. PMID- 15791319 TI - New routes to mesoporous silica-based spheres with functionalized surfaces. AB - Mesoporous hybrid silica-based spheres with functionalized surfaces and abundant highly ordered domains are obtained using for the first time a simple synthetic route based on spray-drying processes. PMID- 15791320 TI - Photocatalytic formation of a carbamate through ethanol-assisted carbonylation of p-nitrotoluene. AB - The nitroarene p-nitrotoluene is converted with a selectivity higher than 85% to the corresponding carbamate at room temperature and atmospheric pressure, using photoexcited particles of TiO2 as catalyst and EtOH as carbonylating species. PMID- 15791321 TI - Two-photon uncaging of neurochemicals using inorganic metal complexes. AB - Neuroactive compounds can be photoreleased by means of two-photon excitation using a new kind of transition metal-based caged compound. PMID- 15791322 TI - Highly efficient conjugate reduction of alpha,beta-unsaturated nitriles catalyzed by copper/xanthene-type bisphosphine complexes. AB - Alpha,beta-unsaturated nitriles are chemoselectively reduced to the corresponding saturated nitriles in high yields using a copper-DPEphos or Xantphos complex as catalyst in the presence of polymethylhydrosiloxane (PMHS) as the stoichiometric reducing agent and t-butanol as additive. PMID- 15791323 TI - Tuning of spin crossover equilibrium in catecholatoiron(III) complexes by supporting ligands. AB - Introduction of electron-withdrawing groups on co-ligands effectively raises the spin crossover temperature of catecholatoiron(III) complexes and induces a significant amount of the low spin species in solution even at around room temperature. PMID- 15791324 TI - Killing three birds with one stone--simultaneous operando EPR/UV-vis/Raman spectroscopy for monitoring catalytic reactions. AB - For the first time, three operando methods, namely EPR, UV-vis and laser-Raman spectroscopy have been applied in parallel on the same V/TiO2 catalyst and under identical reaction conditions during oxidative dehydrogenation of propane to elucidate structure-reactivity relationships in status operandi. PMID- 15791325 TI - The novel and efficient direct synthesis of N,O-acetal compounds using a hypervalent iodine(III) reagent: an improved synthetic method for a key intermediate of discorhabdins. AB - The use of hypervalent iodine(III) reagents allowed us to develop the novel and efficient direct synthesis of N,O-acetal compounds via the oxidative fragmentation reaction of alpha-amino acids or alpha-amino alcohols; furthermore, we succeeded in developing an improved synthesis of the key intermediate of discorhabdins. PMID- 15791326 TI - Flame synthesis of calcium-, strontium-, barium fluoride nanoparticles and sodium chloride. AB - Non-oxidic salts such as NaCl, CaF2, SrF2 and BaF2 were synthesised using a flame spray method; optional doping of such fluorides with rare earth elements suggests possible applications in optics. PMID- 15791327 TI - Tubular assembly of amphiphilic rigid macrocycle with flexible dendrons. AB - An amphiphilic rigid macrocycle was shown to self-assemble into tubular aggregates that can solubilize SWNTs in aqueous solution through significant pi pi interactions. PMID- 15791328 TI - Efficient asymmetric synthesis of an azasugar in water. AB - An extremely efficient asymmetric synthesis of a pyrrolidine azasugar was completed in only four steps in water, without the use of protecting groups and in 60% overall yield from a simple, achiral bis-electrophile. PMID- 15791329 TI - One-step template-directed synthesis of multifunctionalised nanoporous silica: on the way to interactive nanomaterials. AB - The preparation of multifunctional mesoporous silica containing a NLO chromophore in the framework (bridged azobenzene phosphonium salts) and mercaptopropyl groups able to stabilize gold(0) nanoparticles in the channel pores was achieved in one step by using the direct liquid crystal templating approach. PMID- 15791330 TI - Highly active horseradish peroxidase immobilized in 1-butyl-3-methylimidazolium tetrafluoroborate room-temperature ionic liquid based sol-gel host materials. AB - Dramatically enhanced activity and excellent thermal stability of horseradish peroxidase were obtained by immobilizing it in a 1-butyl-3-methylimidazolium tetrafluoroborate room-temperature ionic liquid based sol-gel matrix. PMID- 15791331 TI - Unexpectedly superior enantioselectivity for trans-stilbene cis-dihydroxylation over anchored triosmium carbonyl species in confined Al-MCM-41 channels. AB - Superior enantioselectivity in the dihydroxylation of trans-stilbene catalysed by anchored triosmium carbonyl species without using a chiral modifier is observed inside sterically congested MCM-41 channels; this effect is more pronounced through the introduction of surface Al sites into the silicate. PMID- 15791332 TI - Site directed maleimide bifunctional chelators for the M(CO)3+ core (M =(99m)Tc, Re). AB - A series of bifunctional chelates containing a tridentate donor set for complexation of the M(CO)3+ core and a maleimide group for site-specific coupling to peptides and proteins containing free thiol groups has been prepared and their Re(CO)3+ complexes and glutathione conjugates structurally characterized. The flexibility of design allows preparation of ligands suitable for both fluorescence imaging, radioimaging and radiotherapeutic studies of proteins and peptides as well as other biopolymers using site specific conjugation. PMID- 15791333 TI - Laser induced disruption of bacterial spores on a microchip. AB - We report on the development of a laser based spore disruption method. Bacillus globigii spores were mixed with a laser light absorbing matrix and co crystallized into 200-microm-wide and 20-microm-deep nanovials formed in a polydimethylsiloxane (PDMS) target plate. Surface tension effects were exploited to effect up to 125-fold spore enrichment. When the target zones were illuminated at atmospheric pressure with pulsed UV-laser light at fluences below 20 mJ cm(-2) a change in spore morphology was observed within seconds. Post illumination PCR analysis suggests the release of endogenous DNA indicative of spore disruption. For laser fluences above 20 mJ cm(-2), desorption of spores and fragments was also observed even without a matrix being employed. Desorbed material was collected in a PDMS flowcell attached to the target plate during laser illumination. This opens up a route towards the direct extraction of released DNA in an integrated spore disruption-PCR amplification microchip device. PMID- 15791334 TI - A MALDI-chip integrated system with a monitoring window. AB - The integration of a monitoring port along the microfluidic path of a MALDI-chip integrated device is described. Optimization of the microreactor design allows longer reaction and measuring times. The Schiff base reaction between 4-tert butylaniline (1) and 4-tert-butylbenzaldehyde (2) in ethanol was carried out on chip in the MALDI ionization chamber and the formed imine 3 was detected in real time, demonstrating the feasibility of the "monitoring window" approach. This preliminary result opens the way to on-chip kinetic studies by MALDI-MS, by opening multiple monitoring windows along the microchannel. PMID- 15791335 TI - Toward on-chip X-ray analysis. AB - The possibility of performing chemical analysis and structure determinations with the use of X-rays in a microfluidic chip environment is explored. Externally generated radiation, radioisotope irradiation and on-chip generated X-rays were considered as excitation means for the performance of sample analysis with the techniques of X-ray fluorescence and diffraction. The absorption properties of chip-building materials by different radiation sources are reviewed and data on absorption coefficients calculated, upon which recommendations for optimisations with the use of various X-ray sources may be made. The capabilities and limitations of on-chip X-ray analysis are placed in perspective by preliminary experimental results of diffraction, fluorescence and on-chip X-ray generation experiments. PMID- 15791336 TI - Denaturing gradient-based two-dimensional gene mutation scanning in a polymer microfluidic network. AB - An integrated two-dimensional (2-D) DNA separation platform, combining standard gel electrophoresis with temperature gradient gel electrophoresis (TGGE) on a polymer microfluidic chip, is reported. Rather than sequentially sampling DNA fragments eluted from standard gel electrophoresis, size-resolved fragments are simultaneously electrokinetically transferred into an array of orthogonal microchannels and screened for the presence of sequence heterogeneity by TGGE in a parallel and high throughput format. A bulk heater assembly is designed and employed to externally generate a temporal temperature gradient along an array of TGGE channels. Extensive finite element modeling is performed to determine the optimal geometries of the microfluidic network for minimizing analyte band dispersion caused by interconnected channels in the network. A pH-mediated on chip analyte stacking strategy is employed prior to the parallel TGGE separations to further reduce additional band broadening acquired during the electrokinetic transfer of DNA fragments between the first and second separation dimensions. A comprehensive 2-D DNA separation is completed in less than 5 min for positive detection of single-nucleotide polymorphisms in multiplex PCR products that vary in size and sequence. PMID- 15791338 TI - Effects of the cell geometry and operating parameters on the performance of an external contactless conductivity detector for microchip electrophoresis. AB - Quantitative data on the effect of the electrode geometry on the signal strength and the signal-to-noise ratio is given. The measurements are affected by the unavoidable presence of stray capacitance. Best results are achieved for short and narrow electrodes arranged in an antiparallel configuration and separated by a minimal gap, which determines the dimensions of the actual detection volume. Limits of detection between 150 and 250 microg l(-1) and separation efficiencies from 13,000 to 17,000 theoretical plates were achieved for six inorganic cations (NH(4)(+), K(+), Ca(2+), Na(+), Mg(2+)and Li(+)) with electrodes of 1 mm width and a detection gap of 0.5 mm (separation channel length: 7.5 cm) when operating the detector at 20 V(pp) and 500 kHz. The analyses of all major inorganic cations in tap and rain water samples were demonstrated for the first time in microchip electrophoresis with contactless conductivity detection. PMID- 15791337 TI - Human neural stem cell growth and differentiation in a gradient-generating microfluidic device. AB - This paper describes a gradient-generating microfluidic platform for optimizing proliferation and differentiation of neural stem cells (NSCs) in culture. Microfluidic technology has great potential to improve stem cell (SC) cultures, whose promise in cell-based therapies is limited by the inability to precisely control their behavior in culture. Compared to traditional culture tools, microfluidic platforms should provide much greater control over cell microenvironment and rapid optimization of media composition using relatively small numbers of cells. Our platform exposes cells to a concentration gradient of growth factors under continuous flow, thus minimizing autocrine and paracrine signaling. Human NSCs (hNSCs) from the developing cerebral cortex were cultured for more than 1 week in the microfluidic device while constantly exposed to a continuous gradient of a growth factor (GF) mixture containing epidermal growth factor (EGF), fibroblast growth factor 2 (FGF2) and platelet-derived growth factor (PDGF). Proliferation and differentiation of NSCs into astrocytes were monitored by time-lapse microscopy and immunocytochemistry. The NSCs remained healthy throughout the entire culture period, and importantly, proliferated and differentiated in a graded and proportional fashion that varied directly with GF concentration. These concentration-dependent cellular responses were quantitatively similar to those measured in control chambers built into the device and in parallel cultures using traditional 6-well plates. This gradient generating microfluidic platform should be useful for a wide range of basic and applied studies on cultured cells, including SCs. PMID- 15791339 TI - Parallel nanoliter detection of cancer markers using polymer microchips. AB - A general multipurpose microchip technology platform for point-of-care diagnostics has been developed. Real-time nucleic acid sequence-based amplification (NASBA) for detection of artificial human papilloma virus (HPV) 16 sequences and SiHa cell line samples was successfully performed in cyclic olefin copolymer (COC) microchips, incorporating supply channels and parallel reaction channels. Samples were distributed into 10 parallel reaction channels, and signals were simultaneously detected in 80 nl volumes. With a custom-made optical detection unit, the system reached a sensitivity limit of 10(-6) microM for artificial HPV 16 sequences, and 20 cells microl(-1) for the SiHa cell line. This is comparable to the detection limit of conventional readers, and clinical testing of biological samples in polymer microchips using NASBA is therefore possible. PMID- 15791340 TI - Dynamics of capillary flow of blood into a microfluidic channel. AB - In this paper, a novel mathematical approach is devised to analyze the flow of blood from a droplet into a microcapillary channel. Special attention is devoted to estimate the effects of variable hydraulic resistance over different flow regimes, influence of suspended RBC particulates on the non-Newtonian flow characteristics and implications of a dynamically-evolving contact angle. Flow characteristics depicting advancement of the fluid within the microfluidic channel turn out to be typically non-linear, as per relative instantaneous strengths of the capillary forces and viscous resistances. It is found that the greater the 'pseudoplasticity' of the blood, the weaker the retarding shear forces. The driving forces, on the other hand, become stronger with time, on account of a reduction of contact angle with a decrease of blood flow velocity, although this strengthening is less prominent for blood samples with greater 'pseudoplasticity'. It is revealed that RBCs suspended in blood samples have a strong influence on the effective blood viscosity, and consequently, may drive the fluid significantly faster into the microchannel, especially when the characteristic length scales of the suspensions approach the hydraulic radius of the channel. PMID- 15791341 TI - A microfluidic system enabling Raman measurements of the oxygenation cycle in single optically trapped red blood cells. AB - Using a lab-on-a-chip approach we demonstrate the possibility of selecting a single cell with certain properties and following its dynamics after an environmental stimulation in real time using Raman spectroscopy. This is accomplished by combining a micro Raman set-up with optical tweezers and a microfluidic system. The latter gives full control over the media surrounding the cell, and it consists of a pattern of channels and reservoirs defined by electron beam lithography that is moulded into rubber silicon (PDMS). Different buffers can be transported through the channels using electro-osmotic flow, while the resonance Raman response of an optically trapped red blood cell (RBC) is simultaneously registered. This makes it possible to monitor the oxygenation cycle of the cell in real time and to investigate effects like photo-induced chemistry caused by the illumination. The experimental set-up has high potential for in vivo monitoring of cellular drug response using a variety of spectroscopic probes. PMID- 15791342 TI - Highly sensitive signal detection of duplex dye-labelled DNA oligonucleotides in a PDMS microfluidic chip: confocal surface-enhanced Raman spectroscopic study. AB - Rapid and highly sensitive detection of duplex dye-labelled DNA sequences in a PDMS microfluidic channel was investigated using confocal surface enhanced Raman spectroscopy (SERS). This method does not need either an immobilization procedure or a PCR amplification procedure, which are essential for a DNA microarray chip. Furthermore, Raman peaks of each dye-labelled DNA can be easily resolved since they are much narrower than the corresponding broad fluorescence bands. To find the potential applicability of confocal SERS for sensitive bio-detection in a microfluidic channel, the mixture of two different dye-labelled (TAMRA and Cy3) sex determining Y genes, SRY and SPGY1, was adsorbed on silver colloids in the alligator teeth-shaped PDMS microfluidic channel and its SERS signals were measured under flowing conditions. Its major SERS peaks were observable down to the concentration of 10(-11) M. In the present study, we explore the feasibility of confocal SERS for the highly sensitive detection of duplex dye-labelled DNA oligonucleotides in a PDMS microfluidic chip. PMID- 15791343 TI - Simultaneous measurements of the flow velocities in a microchannel by wide/evanescent field illuminations with particle/single molecules. AB - A laser-induced fluorescence imaging method was developed to simultaneously measure flow velocities in the middle and near wall of a channel with particles or single molecules, by selectively switching from the wide field excitation mode to the evanescent wave excitation mode. Fluorescent microbeads with a diameter of 175 nm were used to calibrate the system, and the collisions of microbeads with channel walls were directly observed. The 175 nm microbeads velocities in the main flow and at 275 nm from the bottom of the channel were measured. The measured velocities of particles or single molecules in two positions in a microchannel were consistent with the calculated value based on Poiseuille flow theory when the diameter of a microbead was considered. The errors caused by Brownian diffusion in our measurement were negligible compared to the flow velocity. Single lambda DNA molecules were then used as a flowing tracer to measure the velocities. The velocity can be obtained at a distance of 309.0 +/- 82.6 nm away from bottom surface of the channel. The technique may be potentially useful for studying molecular transportation both in the center and at the bottom of the channel, and interactions between molecules and microchannel surfaces. It is especially important that the technique can be permitted to measure both velocities in the same experiment to eliminate possible experimental inconsistencies. PMID- 15791344 TI - Laser induced fluorescence photobleaching anemometer for microfluidic devices. AB - We have developed a novel, non-intrusive fluid velocity measurement method based on photobleaching of a fluorescent dye for microfluidic devices. The residence time of the fluorescent dye in a laser beam depends on the flow velocity and approximately corresponds to the decaying time of the photobleaching of the dye in the laser beam. The residence time is inversely proportional to the flow velocity. The fluorescence intensity increases with the flow velocity due to the decrease of the residence time. A calibration curve between fluorescence intensity and known flow velocity should be obtained first. The calibration relationship is then used to calculate the flow velocity directly from the measured fluorescence intensity signal. The new method can measure the velocity very quickly and is easy to use. It is demonstrated for both pressure driven flow and electroosmotic flow. PMID- 15791345 TI - Stacking due to ionic transport number mismatch during sample sweeping on microchips. AB - Sample stacking can occur in isoconductive buffer systems as a result of ion transport mismatches that cause changes in buffer conductivity during electrophoresis. Fluorescence imaging was used to examine this effect in the sweeping of hydrophobic dyes with sodium dodecyl sulfate (SDS) on microchips. Imaging revealed the occurrence of a stacking effect in a sodium borate buffer system in which the sample buffer and SDS-containing run buffer had the same initial conductivity. Injected sample plugs were first swept by SDS micelles and the swept band was then stacked at the trailing end of the sample zone. This effect is due to changes in conductivity at both the front and back interfaces of the injected sample plug and can be modeled by moving boundary equations. Maximum signal enhancements of 86-, 160- and 560-fold were obtained for Rhodamine 560, Rhodamine B and Rhodamine 6G, respectively, by the combination of sweeping and stacking within a 1 cm section of microchannel. Based on sample sweeping/stacking and manipulation of the electric field polarity, a method of trapping and concentrating analyte from multiple injections was also demonstrated. PMID- 15791346 TI - A high current density DC magnetohydrodynamic (MHD) micropump. AB - This paper describes the working principle of a DC magnetohydrodynamic (MHD) micropump that can be operated at high DC current densities (J) in 75-microm-deep microfluidic channels without introducing gas bubbles into the pumping channel. The main design feature for current generation is a micromachined frit-like structure that connects the pumping channel to side reservoirs, where platinum electrodes are located. Current densities up to 4000 A m(-2) could be obtained without noticeable Joule heating in the system. The pump performance was studied as a function of current density and magnetic field intensity, as well as buffer ionic strength and pH. Bead velocities of up to 1 mm s(-1) (0.5 microL min(-1)) were observed in buffered solutions using a 0.4 T NdFeB permanent magnet, at an applied current density of 4000 A m(-2). This pump is intended for transport of electrolyte solutions having a relatively high ionic strength (0.5-1 M) in a DC magnetic field environment. The application of this pump for the study of biological samples in a miniaturized total analysis system (microTAS) with integrated NMR detection is foreseen. In the 7 T NMR environment, a minimum 16 fold increase in volumetric flow rate for a given applied current density is expected. PMID- 15791347 TI - Replica multichannel polymer chips with a network of sacrificial channels sealed by adhesive printing method. AB - Replica microchips for capillary array electrophoresis containing 10 separation channels (50 microm width, 50 microm depth and 100 microm pitch) and a network of sacrificial channels (100 microm width and 50 microm depth) were successfully fabricated on a poly(methyl methacrylate) (PMMA) substrate by injection molding. The strategy involved development of moving mask deep X-ray lithography to fabricate an array of channels with inclined channel sidewalls. A slight inclination of channel sidewalls, which can not be fabricated by conventional deep X-ray lithography, is highly required to ensure the release of replicated polymer chips from a mold. Moreover, the sealing of molded PMMA multichannel chips with a PMMA cover film was achieved by a novel bonding technique involving adhesive printing and a network of sacrificial channels. An adhesive printing process enables us to precisely control the thickness of an adhesive layer, and a network of sacrificial channels makes it possible to remove air bubbles and an excess adhesive, which are crucial to achieving perfect sealing of replica PMMA chips with well-defined channel and injection structures. A CCD camera equipped with an image intensifier was used to simultaneously monitor electrophoretic separations in ten micro-channels with laser-induced fluorescence detection. High speed and high-throughput separations of a 100 bp DNA ladder and phi X174 Hae III DNA restriction fragments have been demonstrated using a 10-channel PMMA chip. The current work establishes the feasibility of mass production of PMMA multichannel chips at a cost-effective basis. PMID- 15791348 TI - Long-term cholinergic treatment is not associated with greater risk of weight loss during Alzheimer's disease: data from the French REAL.FR cohort. AB - BACKGROUND: An increased incidence of weight loss has been reported in patients with Alzheimer's disease (AD) treated with higher doses of acetylcholinesterase inhibitors (AChEI) compared with placebo patients in several clinical trials. The proportion of patients losing weight is extremely variable from one study to another and further analysis is necessary to reach a conclusion on the association of weight loss and AChEI. OBJECTIVE: This observational study was designed to investigate the potential effects of AChEI use on weight loss during AD. DESIGN: 486 patients with AD were followed for one year (initial mean age 77.3 +/- 77 years; initial mean MMS score 20.3 +/- 4.2). Comprehensive geriatric and neuropsychological assessment was conducted every 6 months. Cholinergic treatment was recorded at each visit, as well as any concomitant medication for dementia, psychotropic and other medications. We defined clinically significant weight loss as > or = 4% of the subject's initial weight based on the last measured weight. The data were initially evaluated categorically to identify those who had lost > or = 4% of their initial weight or had remained stable. RESULTS: Eighty-nine per cent of AD patients were treated with AchEI during the first year of follow-up. Twenty-one per cent experienced clinically significant weight loss during this period. Weight loss was associated with more rapid deterioration of cognitive function (Delta MMSE -2.62 +/- 3.99 versus -1.72 +/- 3.64, P = 0.014) and loss of independence in instrumental activities of daily living (Delta IADL -1.45 +/- 1.50 versus - 0.88 +/- 1.43, P = 0.002). The frequency of weight loss was similar whether AD patients were treated with AChEI or not (respectively 21.1 and 19.5%, P = 0.81). In multivariate analysis, the risk of weight loss was significantly decreased in patients taking AChEI for more than 3 months compared (OR = 0.25, 95% CI = 0.11-0.56, P = 0.0007). CONCLUSIONS: Our data suggest that long-term cholinergic treatment is not associated with greater risk of weight loss during AD and may be a protective factor. Further analysis is necessary to confirm this relation. It is obvious that the benefit observed might also be partly related to the organised non-pharmacological management provided for our patients, which included a specific care plan for each individual and facilitated response to nutritional problems as they occurred. Global care of AD patients must associate regular pluridisciplinary management with AChEI treatment. PMID- 15791349 TI - Impact of nutritional status on the evolution of Alzheimer's disease and on response to acetylcholinesterase inhibitor treatment. AB - BACKGROUND: Weight loss is frequently observed in patients with Alzheimer's disease (AD), as observed in clinical practice and reported in the literature. However, information on the evolution of nutritional status and its impact on the prognosis of AD is still scarce. OBJECTIVE: Our aim was to determine the impact of nutritional status on the evolution of AD and on the response to treatment with acetylcholinesterase inhibitors (AChEI) by prospective one-year follow-up of AD patients living at home. METHODS: We studied a cohort of 523 patients with Alzheimer's disease referred from 1994 to 2002 to an Alzheimer centre. After diagnosis, they were followed for one year in a prospective observational study in clinical practice. At entry and every 6 months, patients underwent standardised neurocognitive and geriatric evaluation (MMSE, ADAS-cog, IADL, MNA, caregiver burden). These evaluations were accompanied by complete clinical examination, standard paraclinical investigations and recording of treatment received. RESULTS: Of our patients, 25.8% presented at inclusion a risk of undernutrition with an MNA score of 23.5 or less. During follow-up, the number of patients with rapid loss on the MMSE (3 points or more in one year) was higher in subjects who presented a risk of undernutrition at inclusion (53.6%) than in well nourished subjects (43.2%) (P = 0.07). Similarly, increased dependence at one year was more frequent in subjects at risk of undernutrition at inclusion (57.7% versus 44.4%, P = 0.0219). The beneficial effect of AChEI treatment on cognitive function was not influenced by initial nutritional status; on the contrary, among the subjects at risk of undernutrition at inclusion, the risk of rapid loss on the MMSE in one year was decreased in subjects treated during follow-up compared with untreated subjects (43.9% versus 73.1% ; OR = 0.29; 95% CI = 0.10-0.83; P = 0.0219). This relationship was not found in subjects whose initial MNA score was greater than 23.5. CONCLUSION: Our work indicates that AD patients living at home with a caregiver are frequently at risk of undernutrition. Undernourished patients seem to present more rapid aggravation of the disease, but paradoxically, these patients appear to be those who best respond to AChEI treatment. PMID- 15791350 TI - Nutritional status assessment during Alzheimer's disease: results after one year (the REAL French Study Group). AB - OBJECTIVES: Weight loss and malnutrition are frequent and serious complications of Alzheimer's disease. The aim of the present article was to describe the cognitive and behavioural characteristics of the test population within the frame of the PHRC REAL.FR cohort (for Reseau sur la Maladie d'Alzheimer Francais), depending on their nutritional state, and to consider their evolution one year after the original inclusion. METHOD: The study population' stratification was done in three groups according to their Mini Nutritional Assessment (MNA) score: malnutrition group (MNA < 17.5), at risk of malnutrition group (MNA 17.5-23.5), and normal nutritional status group (MNA > or = 23.5). 561 patients were evaluated at inclusion time, 393 at one year. The evaluation included the following scales: Mini Nutritional Assessment (MNA), Mini Mental State Examination (MMSE), Activities Daily Living (ADL), Instrumental Activities Daily Living (IADL), Neuro Psychiatric Inventory (NPI) and Zarit scale (ZARIT). Comparison and descriptive analysis for each MNA group at baseline and at one year has been performed. RESULTS: at baseline, the well-nourished and the malnutrition risk groups are significantly different concerning age, IADL and NPI; the well-nourished and undernutrition groups are different concerning MMSE, NPI and Zarit; the malnutrition risk and undernutrition groups are only different concerning NPI. At one year, the well-nourished and the malnutrition risk and undernutrition groups are different concerning one lonely variable, the NPI, in a significant way. The comparison of the three groups between baseline and one-year evaluation demonstrate for the well-nourished group an aggravation of MMSE, ADL, IADL, NPI, for the malnutrition risk group of MMSE and IADL, and for the undernutrition group of MMSE, IADL and NPI. CONCLUSION: Among the patients suffering from Alzheimer's disease, the most malnutritioned worsen highly on cognitive and functional capacities. Furthermore, the nutritional aggravation seems strongly linked to behavioural disorders aggravation. The improvement of those disorders has therefore to be part of every improvement strategy applied to the nutritional status of the demented patients. PMID- 15791351 TI - Recent data on the natural history of Alzheimer's disease: results from the REAL.FR Study. AB - OBJECTIVES: This paper aims to present the changes observed in the evolution of Alzheimer's disease (AD) in the cohort REAL.FR after one year by taking account new treatments and improved management. METHODS: Four hundred and ninety-eight patients recruited for the REAL.FR study were followed for one year with a standardized case report filled for each patient every 6 months. Changes in the status of these patients were evaluated on various levels: cognitive, functional, behavioural, global, nutritional, social, medical and caregiver burden. Specific treatments were also recorded. RESULTS: A high proportion of patients received specific treatment for AD throughout the year (86%), mainly acetylcholinesterase inhibitors (AChEI) . As expected we observed statistically significant changes in cognitive function (MMS: -1.93 +/- 3.74, p < 0.0001 and ADAS-cog: +2.40 +/- 3.74, p < 0.0001), an overall loss of autonomy (ADL: -0.56 +/- 1.05, p < 0.0001 and IADL: -1.00 +/- 1.46, p < 0.0001), worsening of behavioral disturbances (NPI: +1.85 +/- 14.83, p=0.0047) and a deterioration of general status (CDRSB: +1.63 +/ 2.55, p< 0.0001). Even if the MNA score decreased not significantly, the loss was close to the threshold of significativity (MNA: -0.31 +/- 3.07, p=0.0531). CONCLUSION: We observed a statistically significant change for the worse in most parameters. However, it appears that this deterioration had been relatively slowed by non-pharmacological management and the specific AD treatments. This resulted in stability or improvement of the condition in 63.4% of patient at 1 year. The management proposed (including prescription of AChEI) seemed to have a real impact on the course of the disease during this first year of follow-up. PMID- 15791352 TI - One-year longitudinal evaluation of neuropsychiatric symptoms in Alzheimer's disease. The REAL.FR Study. AB - Behavioral and Psychological Symptoms are major and frequent manifestations of Alzheimer's Disease (AD). The aim of the present study was to evaluate neuropsychiatric symptoms in the PHRC REAL.FR cohort (for Reseau sur la maladie d'Alzheimer Francais) after one year of evolution. Four hundred and eighty two patients with mild and moderate AD were assessed. A majority of them had significant symptoms at inclusion (85.3 % of subjects with mild AD, 89.7% of patients with a moderate AD). Patients with mild AD had a significant increase of the Neuropsychiatric Inventory (NPI) frequency x severity scores for apathy and aberrant motor behavior. Patients with moderate AD had a significant increase of NPI disinhibition, aberrant motor behavior and sleep disorders scores. The variation of NPI total score at one year correlated positively with change in Zarit's caregiver burden score, independently of global cognitive evolution. After one year, a group of 54 patients were institutionalized in nursing home or long term care unit. When compared to non institutionalized patients, the institutionalized group was characterized at base line by a lower MMSE score, a higher Zarit caregiver burden score, and a higher NPI agitation and disinhibition scores. PMID- 15791353 TI - Predictive factors of rate of loss of autonomy in Alzheimer's disease patients. A prospective study of the REAL.FR Cohort. AB - Alzheimer's disease is characterised by a progressive loss of autonomy in activities of daily living. Many patients lose this autonomy rapidly with dramatic consequences for the patients and their relatives, and for health and social services. The aim of this study was to determine, in a large French cohort of community-living Alzheimer patients (REAL.FR), the factors underpinning different rates of loss of autonomy. Six hundred and eighty seven patients were recruited to this French cohort. Autonomy in activities of daily living was estimated with the IADL scale (Lawton). Patients were divided into three groups according to loss of autonomy during the first one-year follow-up period. Patients with a decrease in the IADL score had worse ADAS-cog score (Alzheimer's Disease Assessment Scale, cognitive subscale), Mini-Mental State Examination (MMSE), and Clinical Dementia Rating Scale scores. Up to a point, a marked decrease in the IADL score was less frequent among patients with a better ADAS cog score (p < 0.10, bilateral test). At one-year evaluation, patients with a marked decline in autonomy were characterised by a greater decrease in Mini Mental State score, faster progression of behavioural disturbance, and more hospitalisations. Further prospective studies, using established models, are needed to isolate the factors associated with a high rate of loss of autonomy in activities of daily living in Alzheimer patients. PMID- 15791354 TI - Evolution of blood pressure in patients with Alzheimer's disease: a one year survey of a French Cohort (REAL.FR). AB - OBJECTIVES: To determine the evolution of blood pressure in patients with moderate Alzheimer's disease among a one year longitudinal survey and to evaluate the relationship between blood pressure and cognitive functions. METHODS: In 327 subjects selected from the French research program on Alzheimer's disease (REAL.FR), systolic and diastolic blood pressure (SBP, DBP) were measured at the time of inclusion (M0), after 6 months (M6) and after 12 months (M12). All subjects were assessed to determine both cognitive functions and capabilities in the activities of daily living using validated cognitive scales [Mini Mental State Examination (MMSE), Alzheimer's Disease Assessment Scale--Cognitive part (ADAS-Cog), Instrumental Activities of Daily Living (IADL), Activities of Daily Living (ADL), Clinical Dementia Rating (CDR)], at M0, M6, M12. RESULTS: In this population of patients with moderate Alzheimer's disease, mean age was 78 +/- 7 years and 242 subjects were females (74%). After adjustment for age, gender, body mass index (BMI) and antihypertensive therapy, a significant decrease of blood pressure was observed between M0 and M12, for SBP (139.1 +/- 18 to 136.5 +/- 17 mmHg, p < 0.05) and DBP, (77.6 +/- 12 to 75.8 +/- 10 mmHg , p < 0.05). Demented subjects with the worst cognitive impairment at baseline (tertile1 MMSE, tertile 3 ADAS-Cog, ADL scores between 0 and 4, CDR scores between 10 to 18) showed a larger decrease in SBP and DBP after 12 months. The worst impairment in dementia at baseline was associated with the highest SBP decrease between M0 and M12 (delta SBP tertile 1 MMSE vs tertile 3 MMSE = -5.9 vs + 1.0 mmHg , p < 0.05; Delta SBP tertile 3 ADAS-Cog vs tertile 1 ADAS-Cog = - 5.98 vs + 2.98 mmHg, p < 0.05, Delta SBP ADL 0-4 vs ADL -6 = - 8.7 vs -1.5 mmHg, p < 0.05, delta SBP CDR 10-18 vs CDR 0.5-9.5 = - 6.9 vs -1.7 mmHg, p < 0.05). All these results persisted after adjustment for age, gender and the antihypertensive therapy. Baseline SBP [OR 95% CI = 1.05 (1.02-1.08), BMI [OR 95% CI = 0.88 (0.81-0.95)], ADL score [OR 95% CI = 0.42 (0.22-0.81)] and ADAS-Cog score [OR 95% CI = 1.07 (1.01-1.14)] were significantly associated with the decrease of blood pressure after one year of follow up, independently of age, gender and antihypertensive therapy. In contrast, patients with larger blood pressure decrease (over 10 mmHg reduction of SBP and/or 5 mmHg of DBP) did not demonstrate a more significant worsening of dementia at 12 months in the different scales used. CONCLUSIONS: This study indicates a significant decrease in blood pressure in patients with Alzheimer's disease after one year of follow up, independently of age, gender, BMI and antihypertensive therapy. The largest decrease in blood pressure was observed in patients with the most severe impairment in dementia at baseline, suggesting that blood pressure decrease seems to be mainly a secondary phenomenon in Alzheimer's disorders. PMID- 15791355 TI - Predictive factors of emergency hospitalisation in Alzheimer's patients: results of one-year follow-up in the REAL.FR Cohort. AB - OBJECTIVE: To determine the predictive factors of hospitalisation in patients with Alzheimer's disease followed in the REAL.FR cohort. METHODS: A French multicentre prospective study (REAL.FR) following 516 patients who had mild to moderate AD at inclusion. Analysis of the data after one year of follow-up. RESULTS: 139 (26,9%) of the 516 initial AD patients were hospitalized during the 1 year of follow-up. After bivariate analysis, the principal predictive factors of hospitalisation were high scores on the Reisberg scale (> or = 5: P = 0.0149) and the CDR (1: P = 0.0289; 2 or 3: P = 0.0078); > or = 2 intercurrent diseases (P = 0.00104); > or = 3 other treatments (not including specific treatments for AD) (P = 0.0026); BMI (kg/m2) between 25 and 30 (P = 0.0147); impossibility of single-leg stance (P = 0.02); > or = 1 disabilities on the ADL (P = 0.0009) and > or = 2 disabilities on the IADL (P = 0.0017); use of medical services (P = 0.0236) and of non-medical services (P = 0.0403); delirium or hallucinations (P = 0.0135), depression (P = 0.0014), or disinhibited behaviour (P = 0.0030); the gravity and frequency of behavioural symptoms (NPI freq x grav > or = 11 (median), P = 0.0012); and lastly, a score of > or = 20 for subjective caregiver burden on the Zarit scale (P < 0.0001). Multivariate analysis revealed an association between the risk of hospitalisation and the following variables : the type of centre to which the patient was admitted (neurological, psychiatric or geriatric), impaired orientation on the MMS, BMI, the number of disabilities on the ADL, and caregiver burden as evaluated by the Zarit scale. CONCLUSION: At inclusion, patients with more severe cognitive disorders, poor nutritional status and those who were the most dependent for basic activities of daily living were already at greater risk of hospitalisation. Exhaustion of the informal caregiver was an independent and supplementary predictive factor of hospitalisation. PMID- 15791356 TI - Living alone with Alzheimer's disease: cross-sectional and longitudinal analysis in the REAL.FR Study. AB - OBJECTIVES: The purpose of this study was to examine the characteristics of Alzheimer's disease (AD) patients living alone and to describe the rate of cognitive and functional impairment after a one-year follow-up. DESIGN AND SETTING: In a prospective longitudinal study conducted by the French network on Alzheimer's disease (the REAL.FR study), 677 older community-dwelling AD patients were interviewed and completed questionnaires and evaluation scales every 6 months during a one-year follow-up. MEASUREMENTS: All patients were assessed by trained staff who collected data on neuropsychological status using the Mini Mental State Examination (MMSE), behavioural disturbances with the Neuropsychiatric Inventory (NPI) and nutritional status with the Mini Nutritional Assessment (MNA). Patients were assessed for current mobility and function in activities of daily living (ADL) and instrumental activities of daily living (IADL). RESULTS: At inclusion, 28% of the 677 non-institutionalised individuals with AD lived alone. Those who lived alone were significantly older than those who did not, and among them the percentage of women was significantly higher. Patients living alone were at increased risk of malnutrition and were more likely to have a low income than those living with others. Persons with AD living alone made greater use of health services. Dementia stage evaluated by cognitive impairment (MMSE) and ADL disabilities was similar in both groups. At one-year follow-up, the mortality rate was significantly higher in AD patients living with others. Institutionalisation and hospitalisation rates were similar. CONCLUSION: These results draw attention to the fact that elderly persons with AD living alone are a subpopulation with specific needs which require the development of targeted interventions. Further investigation of the factors associated with the lower mortality rate in AD patients living alone is necessary, and the results of long-term follow-up in this prospective study should shed light on this question. PMID- 15791357 TI - Changes in informal care over one year for elderly persons with Alzheimer's disease. AB - BACKGROUND: Informal caregivers play a major role in all types of assistance for elderly persons with Alzheimer's disease but few longitudinal studies reports change in this role over time. OBJECTIVE: The aim of our research was to describe the objective and subjective burden of informal caregivers of elderly persons with dementia of Alzheimer type, and to follow its evolution during one year. METHODS: A multicentre prospective study of 333 principal caregivers of patients with Alzheimer's disease, followed for one year. RESULTS: At inclusion, the majority of caregivers were involved in most tasks of assistance (activities of daily living, handling money, supervision, organisation of support services) but only 13.4% were involved in basic activities of daily living. As expected, during follow-up, the involvement of caregivers increased and extended to all tasks: whereas at inclusion 11.0% regularly assisted in all tasks, after only 12 months follow-up this figure rose to 28.9% (P < 0.001). Whereas 45.9% of caregivers became involved in carrying out new tasks (mainly organisation of support services and help with basic activities of daily living), 8.9% no longer carried out certain tasks (essentially supervision and organisation of support services), 7.3% replaced one task by another, and for 37.8% there was no change. During the same time, the mean burden experienced by the caregiver showed a very slight change (increasing from a mean score of 21.08 +/- 14.65 to 22.68 +/- 16.45, P = 0.044) with great variation between caregivers. Those who increased their involvement also had a significant increase in caregiver burden. CONCLUSION: While home caregiving appeared to follow an even course, the needs of the care recipient changed rapidly over time and caregivers differed in their response to these changes, indicating that regular follow-up is required. PMID- 15791359 TI - Endoluminal and transluminal surgery: no longer if, but when. PMID- 15791360 TI - Surgical education and training in the new millennium. PMID- 15791362 TI - Laparoscopic drainage of pancreatic pseudocysts. AB - BACKGROUND: Laparoscopic drainage of pancreatic pseudocysts (PPs) has been used in selected cases. The aim of this study is to analyze our results with the laparoscopic technique and to compare them with those of a cohort of patients treated by open surgery during the same time period. PATIENTS AND METHODS: Ten patients underwent laparoscopic drainage of PPs during a 7-year period [laparoscopic group (LG)]. The type of drainage was chosen according to the size and location of the PP. Demography, surgical details, results, and complications were analyzed and contrasted with those of 6 patients who underwent open drainage [open group (OG)]. RESULTS: All patients presented with mature PPs developed after a documented episode of acute pancreatitis. Mean age of the LG was 42 years (six males and four females). In the OG, mean age was 36 years (five males and one female). Etiology of the pancreatitis was alcoholic in eight patients, biliary in five, toxic in two, and associated with systemic lupus erythematous in one. Laparoscopic procedures included Roux-en-Y cystojejunostomy in four patients, extraluminal cystogastrostomy in four, and intraluminal cystogastrostomy in two. There were no conversions. In the OG, cystogastrostomy was performed in three patients and Roux-en-Y cystojejunostomy in three. One patient in the LG developed upper gastrointestinal bleeding the day after surgery that resolved uneventfully, one patient presented a postoperative abscess that required open drainage, and one patient presented a residual pseudocyst that was treated by endoscopy. Morbidity in the OG included a small bowel obstruction secondary to an internal hernia that required reoperation, pneumonia, and a residual pseudocyst that was treated conservatively in one patient each. At a median follow-up of 22 months (range, 1-72) all patients were asymptomatic with no evidence of recurrent disease by computed tomography scan. CONCLUSION: Laparoscopic drainage of PPs is feasible, safe, and effective. Results are similar to those obtained using the open technique. PMID- 15791361 TI - The net immunologic advantage of laparoscopic surgery. AB - The trauma of surgery evokes a variety of physiologic and immunologic alterations that should contribute to host defense. However, an exaggerated response to injury may result in immunosuppression and lead to significant postoperative morbidity and mortality. Laparoscopic surgery may result in less induced surgical trauma than conventional open surgery. Decreased postoperative pain and speedy functional recovery of laparoscopic patients may be attributable to the reduced inflammatory response and minimal immunosuppression. Inflammation, an early protective homeostatic immune response to injury, is characterized by the production of proinflammatory cytokines and by activation of cellular and humoral immune mechanisms. Postoperative levels of the inflammatory cytokines have been consistently lower after laparoscopic procedures, indicating a smaller degree of surgical insult and acute inflammatory reaction. Surgical stress derails the functions of both polymorphonuclear and mononuclear cells, which may lead to an increased risk of postoperative infection. Comparative studies of cellular immunity after laparoscopic and conventional surgery demonstrate immunologic advantage conferred by laparoscopy. Exaggerated activation of peritoneal immunity may lead to a relative local immunosuppression, resulting in ineffective intraperitoneal bacterial clearance and serious postoperative infections. Functions of the peritoneal macrophages are better preserved when laparotomy is avoided. Decreased perioperative stress may be particularly important for oncologic patients. Laparoscopic approaches may result in diminished perioperative tumor dissemination and better cancer outcomes. Although laparoscopy is "minimally invasive," systemic immune responses still are undeniably activated. However, laparoscopic surgery appears to induce a smaller injury, resulting in proportionally decreased immunologic changes. In addition to improved cosmesis and faster functional recovery, a patient undergoing laparoscopic surgery may benefit most from a net immunologic advantage. PMID- 15791363 TI - Laparoscopic distal pancreatectomy. AB - BACKGROUND: Advanced laparoscopic techniques have been adapted to various surgical pathologies, including pancreatic tumors, with the potential benefits of attenuated surgical trauma, faster recovery, and improved cosmesis. Laparoscopic pancreatic surgery is technically demanding, and thus has not yet gained widespread acceptance. The aim of this study was to review our preliminary experience with laparoscopic distal pancreatectomy for benign and malignant pancreatic pathologies. METHODS: A retrospective chart review of consecutive patients with benign and malignant pancreatic tumors who underwent laparoscopic distal pancreatectomy in a university-affiliated department of surgery between 1997 and 2003 was performed. Data relative to demographic and clinical characteristics, indications for surgery, surgical procedure, and postoperative course were recorded. RESULTS: Laparoscopic distal pancreatectomy was attempted for 12 patients with benign (n = 8) and malignant (n = 4) pancreatic tumors and successfully completed laparoscopically in 75%, of these cases. Six early postoperative complications (two abscesses, two instances of diabetes mellitus, two pancreatic leaks) developed in three patients. The spleen was successfully preserved in 58% of the cases. CONCLUSIONS: This preliminary experience suggests that laparoscopic distal pancreatectomy is a feasible and safe procedure with a morbidity rate comparable with that for the conventional open procedure. However, laparoscopic surgery for malignant pancreatic tumors remains controversial. Larger series with longer follow-up periods are necessary to determine the role of laparoscopic surgery in the treatment of pancreatic pathologies. PMID- 15791364 TI - Role of pancreatic duct stenting in the treatment of chronic pancreatitis. AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography and stent placement are relatively new alternatives to surgery for the treatment of chronic pancreatitis. The objective of this study was to determine the efficacy of pancreatic duct stent placement for the treatment of chronic pancreatitis. METHODS: This study included 89 patients treated with pancreatic stents between 1993 and 2002. The patients were contacted via telephone for a personal interview with regard to pain, medication usage, weight loss or gain, and eating patterns. Additionally, medication usage before and after treatment was documented from the Kentucky Cabinet for Health Services' electronic reporting system for narcotic use. RESULTS: Of the 89 patients, 9 were deceased, 5 either refused to interview or could not be contacted, and 75 were interviewed. Significant weight gain exceeding 15 lb after treatment was experienced by 22%, whereas only 4% lost weight. A majority of the patients (68%) noted that they had less severe relapses or no relapses after treatment. The patients reported a decrease in pain level on a 10-point scale from 8.7 to 4.1 (53% decrease) after treatment. A decrease in pain medication usage was reported by 47% of the patients, and 83% considered their treatment successful. The Kentucky All Schedule Prescription Electronic Report (KASPER) was obtained before and after treatment for 55 patients. According to this statewide electronic reporting system, 63% had a documented decrease in narcotic use. CONCLUSION: The findings of this study support the use of pancreatic duct stenting as an option before surgical intervention for these difficult-to-manage patients with chronic pancreatitis. PMID- 15791365 TI - Perductal electrolytic ablation of the porcine pancreas: a minimally invasive option-studies of morbidity and mortality. AB - BACKGROUND: Pancreatic cancer has a dismal prognosis. Few patients are suitable for surgical resection, leaving the majority requiring symptom palliation. Current palliative techniques such as surgical bypass and endoscopic retrograde cholangiopancreatography (ERCP) are imperfect. A novel palliative therapy combining the symptom control of surgical bypass with the minimally invasive nature of ERCP is required. METHODS: Perductal electrolytic ablation of pancreatic tissue, in a porcine model, was performed. There were two survival groups of 2 weeks (n = 4) and 8 weeks (n = 4). Postoperatively, serum biochemistry, amylase and C-reactive protein (CRP) were assessed. Histological examination of the pancreas, lungs, and kidneys was performed to determine the presence of acute pancreatitis or systemic inflammatory response. RESULTS: An immediate transient increase in both amylase and CRP was seen. Although pancreatic histology demonstrated localised necrosis at the electrolytic site at 2 weeks, there was no evidence of generalized pancreatitis or a systemic inflammatory response at either 2 or 8 weeks. CONCLUSIONS: This study suggests that, although there is localized pancreatic necrosis and transient hyperamylasemia, perductal pancreatic electrolytic ablation is safe, with neither generalized pancreatitis nor a systemic inflammatory response, in the medium and long term. Although performed in normal porcine pancreas, because of the absence of a large-animal model of pancreatic cancer, this study suggests that electrolytic pancreatic ablation is safe. This technique may have a role in the palliation of pancreatic cancer, especially if delivered via a minimally, invasive approach, and warrants further investigation. PMID- 15791366 TI - Endoscopic sphincterotomy for prevention of the recurrence of acute biliary pancreatitis in patients with gallbladder in situ: long-term follow-up of 88 patients. AB - BACKGROUND: Endoscopic sphincterotomy without cholecystectomy is a therapeutic option in selected patients after acute biliary pancreatitis. We conducted a prospective evaluation of the long-term effects of sphincterotomy in terms of the need for of subsequent cholecystectomy and the recurrence of gallstone pancreatitis. METHODS: We studied 88 patients with acute biliary pancreatitis and an intact gallbladder who, underwent endoscopic sphincterotomy either because they were high-risk candidates for surgery or because they had refused of cholecystectomy. The median follow-up was 51 months (range, 5-86). RESULTS: Only two patients (2.2%) experienced recurrent pancreatitis. Subsequent cholecystectomy was performed in 10 patients because of acute cholecystitis in eight cases and biliary colic in two cases. Sixty-six patients (75%) remained asymptomatic. CONCLUSIONS: Endoscopic sphincterotomy is a safe and acceptable alternative to cholecystectomy for the prevention of recurring attacks of gallstone pancreatitis. As a result of this procedure, 75% of patients remained free of symptoms in the long term. PMID- 15791367 TI - A comparison of the complication rates between laparoscopic colectomy and laparoscopic low anterior resection. AB - BACKGROUND: This study compared the short-term outcomes, including the complication rate and minimum surgical invasiveness, between patients with colon and rectal carcinomas, who underwent laparoscopic surgery. METHODS: A review evaluated 151 patients who underwent laparoscopic colectomy (Lap-colectomy; n = 120) and laparoscopic low anterior resection (Lap-LAR; n = 31) between July 2001 and December 2003. The short-term outcomes were compared between the two groups. RESULTS: The mean operative time and blood loss were significantly greater in the Lap-LAR group. However, the complication rates and postoperative course between the two approaches were similar, and no anastomotic leakage was observed. There was no significant difference in the serum C-reactive protein level and white blood cell count between the two groups in the early postoperative period. CONCLUSIONS: Lap-LAR for rectal carcinoma can be performed safely without increased morbidity or mortality, and its short-term benefits are comparable with those conferred by Lap-colectomy. PMID- 15791368 TI - Laparoscopic colorectal surgery in obese and nonobese patients: do differences in body mass indices lead to different outcomes? AB - BACKGROUND: The aim of this prospective study was to compare the outcome of laparoscopic colorectal surgery in obese and nonobese patients. METHODS: All patients who underwent laparoscopic surgery for both benign and malignant disease within the past 5 years were entered into the prospective database registry. Body mass index (BMI; kg/m(2)) was used as the objective measure to indicate morbid obesity. Patients with a BMI >30 were defined as obese, and patients with a BMI <30 were defined as nonobese. The parameters analyzed included age, gender, comorbid conditions, diagnosis, procedure, duration of surgery, transfusion requirements, conversion rate, overall morbidity rate including major complications (requiring reoperation), minor complications (conservative treatment) and late-onset complications (postdischarge), stay on intensive case unit, hospitalization, and mortality. For objective evaluation, only laparoscopically completed procedures were analyzed. Statistics included Student's t test and chi-square analysis. Statistical significance was assessed at the 5% level (p < 0. 05 statistically significant). RESULTS: A total of 589 patients were evaluated, including 95 patients in the obese group and 494 patients in the nonobese group. There was no significant difference in conversion rate (7.3% in the obese group vs 9.5% in the nonobese group, p > 0.05) so that the laparoscopic completion rate was 90.5% (n = 86) in the obese and 92.7% (n = 458) in the nonobese group. The rate of females was significantly lower among obese patients (55.8% in the obese group vs 74.2% in the nonobese group, p = 0.001). No significant differences were observed with respect to age, diagnosis, procedure, duration of surgery, and transfusion requirements (p > 0.05). In terms of morbidity, there were no significant differences related to overall complication rates with respect to BMI (23.3% in the obese group vs 24.5% in the nonobese group, p > 0.05). Major complications were more common in the obese group without showing statistical significance (12.8% in the obese group vs 6.6% in the nonobese group, p = 0.078). Conversely, minor complications were more frequently documented in the nonobese group (8.1% in the obese group vs 15.5% in the nonobese group, p = 0.080). In the postoperative course, no differences were documented in terms of return of bowel function, duration of analgesics required, oral feeding, and length of hospitalization (p > 0.05). CONCLUSION: These data indicate that laparoscopic colorectal surgery is feasible and effective in both obese and nonobese patients. Obese patients who are thought to be at increased risk of postoperative morbidity have the similar benefit of laparoscopic surgery as nonobese patients with colorectal disease. PMID- 15791369 TI - Laparoscopic-assisted approach in rectal cancer patients: lessons learned from >200 patients. AB - BACKGROUND: The applicability of laparoscopic surgery in the treatment of colorectal diseases is still controversial. Early reports on laparoscopic assisted colectomy in patients with colon cancer suggested that it minimizes surgical trauma, decreases perioperative complications, and leads to a more rapid recovery. To our knowledge, no previous studies have compared the laparoscopic vs the open approach in rectal cancer. The aim of this paper was to assess the results of laparoscopic techniques in patients with rectal cancer. METHODS: From March 1998 to February 2003, all patients admitted to our unit with adenocarcinoma of the rectum were evaluated for surgery by the laparoscopic approach. RESULTS: A total of 220 patients with a mean age of 67.3 years were included in the study. One hundred thirty patients (59%) were treated with neoadjuvant chemoradiotherapy. In >75% of the patients, a surgical procedure with sphincter preservation was performed. The rate of conversion to the open approach was 20%. Ten patients had intraoperative complications. Fifty-eight patients (26.3%) developed postoperative complications. The length of hospital stay was 6.8 days. The distribution of tumor stages was as follows: stage I, 16.81%; stage II, 33.6%; stage III, 26.36%; stage IV, 19.09%. The mean number of lymph nodes was 13.8. The incidence of local relapse was 5.3%, with a follow-up of 18 months. CONCLUSION: Laparoscopic surgery can be safely performed in patients with adenocarcinoma of the rectum with good short-term results. Randomized controlled trials are needed to confirm these results. PMID- 15791370 TI - 'Fast-track' multimodal rehabilitation program improves outcome after laparoscopic sigmoidectomy: a controlled prospective evaluation. AB - BACKGROUND: Laparoscopic colorectal resection improves patient outcome by reducing pain, postoperative pulmonary dysfunction, gastrointestinal paralysis, and fatigue. A multimodal rehabilitation program ("fast-track") with epidural analgesia, early oral feeding, and enforced mobilization may further improve the excellent results of laparoscopic colorectal resection, enabling early ambulation of these patients. METHODS: Fifty two consecutive patients underwent laparoscopic sigmoidectomy with standardized regular perioperative treatment (standard) or multimodal rehabilitation program ("fast-track"). Outcome measures included pulmonary function, duration of postoperative ileus, pain perception, fatigue, morbidity, and mortality. RESULTS: Twenty nine standard-care patients (19 men and 10 women) and 23 fast-track patients (15 men and eight women) were evaluated. Demographic and operative data were similar for the two groups. On the 1st postoperative day, pulmonary function was improved (p = 0.01) in fast-track patients. Oral feeding was achieved earlier (p < 0.01) and defecation occurred earlier (p < 0.01) in the fast-track group. Visual analogue scale scores for pain were similar for the two groups (p > 0.05), but fatigue was increased in the standard-care group on the 1st (p = 0.06) and 2nd (p < 0.05) postoperative days. Morbidity was not different for the two groups. Fast-track patients were discharged on day 4 (range, 3-6) and standard-care patients on day 7 (range, 4 14) (p < 0.001). CONCLUSION: Multimodal rehabilitation can improve further on the excellent results of laparoscopic sigmoidectomy and decrease the postoperative hospital stay. PMID- 15791371 TI - Laparoscopic resection of gastrointestinal mesenchymal tumors located in the upper stomach. AB - BACKGROUND: The treatment strategy for a gastrointestinal mesenchymal tumor located close to the esophagogastric junction remains controversial. The authors evaluate the criteria indicating that a gastrointestinal mesenchymal tumor is suitable for laparoscopic resection and assess the surgical techniques on the basis of clinical outcomes. METHODS: The criteria specified a tumor more than 2 cm in diameter or a tendency for it to increase in size during the follow-up period. For eight patients in whom the tumor was located within 3 cm of the esophagogastric junction, an intragastric laparoscopic approach was used, whereas for seven patients in whom the tumor was further from the esophagogastric junction, an exogastric approach was used. RESULTS: In all 15 cases, the laparoscopic resection was successful, with no complications. The intragastric group had a mean maximal tumor size of 2.9 cm, a mean operation time of 168 min, and a mean postoperative hospital stay of 8.8 days, whereas these values in the exogastric group, were respectively, 3.9 cm, 121 min (p = 0.0442), and 9.6 days. There were no recurrences in either group during the follow-up period. CONCLUSION: The good clinical outcomes suggest hat the criteria used as an indication for laparoscopic resection and the surgical techniques applied were appropriate for the resection of gastrointestinal mesenchymal tumors. PMID- 15791372 TI - Long-term follow-up study of the Stretta procedure for the treatment of gastroesophageal reflux disease. AB - BACKGROUND: The endoscopic delivery of temperature-controlled radiofrequency energy to the gastroesophageal junction (Stretta procedure) recently has been shown effective for patients with gastroesophageal reflux disease (GERD). However, its effectiveness has been assessed mainly over short periods (6-12 months). This study aimed to evaluate long-term results of the Stretta procedure. METHODS: All patients undergoing the Stretta procedure since August 2000 were prospectively evaluated under an institutional review board-approved protocol. All patients with a follow-up period longer than 18 months were recruited for a 24-h pH study and mailed a follow-up survey, which included the following: Short Form 12 (SF-12) health status questionnaire, GERD-specific quality-of-life questionnaire (QOLRAD), and queries regarding long-term satisfaction and medication use. RESULTS: The Stretta procedure was performed on 82 patients, and 41 patients with a follow-up period longer than 18 months qualified for the study. Follow-up surveys were completed by 36 patients (88%) during a mean follow up period of 27.1 +/- 3.7 months. Of these 36 patients, 30 (83%) were highly satisfied with the procedure and would have it performed again. More than half of the Fifty Stretta patients (56%) had completely discontinued their use of proton pump inhibitors (PPIs), and an additional 31% had reduced their dose significantly. The mean PPI equivalent doses were 37.8 +/- 22.2 mg/day before the Stretta procedure and 11.6 +/- 14.6 mg/day at 27-month follow-up assessment (p = 0.001). According to the patient outcomes for daily PPI use (yes/no), the patients were divided into two groups: responders (n = 20) and nonresponders (n = 16). The responder group scored higher in QOLRAD score (p = 0.0001), SF-12 physical score (p = 0.038), and SF-12 mental score (p = 0.003). In the 24-hour pH study, the responder group demonstrated a significant decrease in distal esophageal acid exposure time (6.4% +/- 1.5% to 3.1% +/- 1.4%; p = 0.0001). CONCLUSION: The Stretta procedure results in a statistical significant long-term decrease in GERD symptoms and PPI use. The treatment effect is durable beyond 2 years, and 56% of patients had discontinued their user of all antisecretory drugs. PMID- 15791373 TI - Minilaparoscopic (needlescopic) cholecystectomy: a study of 1,011 cases. AB - BACKGROUND: The safety and feasibility of minilaparoscopic cholecystectomy has not been documented with a large patient sample. This study reports the results of 1,011 minilaparoscopic cholecystectomies performed in a single institution. METHODS: From November 1997 to May 2002, 1,023 consecutive patients underwent minilaparoscopic cholecystectomy at National Taiwan University Hospital, Taipei, Taiwan. Patients with clinical evidence of common bile duct stones (1 patient) and combined surgery for other purposes (11 patients) were excluded. The operative indication, total operative time, conversion rate, hospital stay, morbidity and mortality of 1,011 patients were reviewed and statistically analyzed. RESULTS: Minilaparoscopic cholecystectomy was performed in 1,009 of 1,011 patients (375 males and 636 female; mean age, 54.8 years; range 13-92 years). The total operative time was 68.8 +/- 31.9 min. The total hospital stay was 2.5 +/- 2 days. One patient (0.10%) underwent conversion to open cholecystectomy because of common hepatic duct laceration. One patient (0.10%) underwent conversion to standard laparoscopic cholecystectomy for control of cystic artery bleeding. Ten patients (0.99%) experienced major complications including intraabdominal abscess (1 patient), bile leakage (5 patients), major bile duct injury (2 patients), bowel injury (1 patient), and postoperative hemorrhage (1 patient). Eleven patients (1.09%) had minor complications including wound infection, incisional herniation, postoperative ileus, and acute urine retention. One patient (0.10%) with bleeding tendency succumbed to postoperative hemorrhage. CONCLUSIONS: Minilaparoscopic cholecystectomy is a technically demanding approach. Our results indicate that this procedure could be performed successfully and safely by experienced surgical teams. PMID- 15791374 TI - Diagnostic laparoscopy for penetrating injuries in the thoracoabdominal region. AB - BACKGROUND: The role of laparoscopy in diagnosis of penetrating abdominal injuries is still controversial. In the present investigation diagnostic laparoscopy was studied in penetrating injuries of the thoracoabdominal region. METHODS: Between March 1998 and June 2003, 43 patients with penetrating thoracoabdominal injuries underwent diagnostic laparoscopy at the Rambam Medical Center. There were 41 males and two females; the average age was 30 years (range, 16-54 years). Thirty-one patients had a lower chest injury, eight patients had an upper abdomen and flank injury, and four patients had combined chest and abdomen injuries. In 11 patients intraperitoneal penetration was diagnosed. In 10 patients the procedure was converted to open laparotomy, and one patient with a small laceration of the right diaphragm opposite the liver was observed without laparotomy. RESULTS: The average operating time for the laparoscopy was 25 min (10-45 min), and 85 min (40-175 min) for laparotomy. Patients who underwent laparoscopy were discharged after an average of 1.6 (1-3) days, while those who underwent laparotomy were discharged after an average of 7.6 (2-15) days. CONCLUSIONS: Laparoscopy is a useful diagnostic tool in penetrating injuries of the chest, thoracoabdominal region, and flank. This procedure is particularly reliable in diaphragmatic tears. Laparoscopy should be considered the procedure of choice for the evaluation of penetrating injuries of the lower chest and upper abdomen for diagnosis of peritoneal penetration. PMID- 15791375 TI - Minimally invasive incisional herniorrhaphy: a review of 208 cases. AB - BACKGROUND: Minimally invasive incisional herniorrhaphy has become an accepted approach for incisional hernia. However, the ideal technique for this procedure is not known. The authors present their technique and personal experience with minimally invasive incisional herniorrhaphy. METHODS: A retrospective review investigated 208 consecutive patients who underwent minimally invasive incisional hernia repair under the supervision of a single surgeon between 1995 and 2002. RESULTS: An intraperitoneal mesh repair was performed in all cases. There were no conversions. The mean operative time was 2.1 h (range, 0.8-4.5 h). The mean length of hospital stay was 2.5 days (range, 0-13 days). There were six complications, including two bowel perforations, and zero mortality. There were three recurrences during the follow-up period, which ranged from 6 to 72 months (median, 24 months). CONCLUSIONS: Minimally invasive incisional herniorrhaphy yielded an acceptable morbidity and recurrence rate during the follow-up period. The outcome compares favorably with that for open incisional hernia repair. Although long-term follow-up evaluation is desirable, the data support the contention that the minimally invasive approach is an appropriate option for incisional hernia. PMID- 15791376 TI - Complete versus assisted thoracoscopic approach: a prospective randomized trial comparing a variety of video-assisted thoracoscopic lobectomy techniques. AB - BACKGROUND: Video-assisted thoracoscopic surgery (VATS) lobectomy does not represent a unified approach, but rather a spectrum of operative techniques ranging from a complete endoscopic thoracotomy to a minithoracotomy. A prospective randomized trial was conducted to compare the differences in these techniques and their results to determine the best of VATS lobectomy for lung cancer. METHODS: This study randomized 39 consecutive patients with clinical stage I lung cancer to undergo either a complete (C-VATS, n = 20) or an assisted (A-VATS, n = 19) VATS approach for pulmonary lobectomy. RESULTS: The operating time was longer (p = 0.002) and blood loss was less (p = 0.004) with C-VATS than with A-VATS. Although there was no significant difference in analgesic use or duration of thoracic drainage between the groups, a shorter hospitalization was observed after C-VATS. Serum peak levels of postoperative inflammatory markers (white blood cell count, C-reactive protein, creatine phosphokinase) were lower with C-VATS and an earlier return to normalization than with A-VATS. CONCLUSION: Various differences exist among the VATS lobectomy techniques, and complete VATS lobectomy as a purely endoscopic surgery may be technically feasible and a satisfactory alternative to the conventional procedure for stage I lung cancer. PMID- 15791377 TI - Carbon dioxide pneumoperitoneum causes severe peritoneal acidosis, unaltered by heating, humidification, or bicarbonate in a porcine model. AB - BACKGROUND: Carbon dioxide (CO(2)) is the most common gas used for insufflation in laparoscopy, but its effects on peritoneal physiology are poorly understood. This study looks at the changes in peritoneal and bowel serosal pH during CO(2) pneumoperitoneum, and whether heating and humidification with or without bicarbonate alters the outcomes. METHODS: Twenty-one pigs divided into four groups as follows: (1) standard (STD) laparoscopy (n = 5); (2) heated and humidified (HH) laparoscopy (n = 6); (3) heated and humidified with bicarbonate (HHBI) laparoscopy (n = 5); and (4) laparotomy (n = 5). Peritoneal pH, bowel serosal pH, and arterial blood gas (ABG) were obtained at 15-min intervals for 3 h. RESULTS: Severe peritoneal acidosis (pH range 6.59-6.74) was observed in all laparoscopy groups, and this was unaltered by heating and humidification or the addition of bicarbonate. Bowel serosal acidosis was observed in all laparoscopy groups with onset of pneumoperitoneum, but it recovered after 45 minutes. No significant changes in peritoneal or bowel serosal pH were observed in the laparotomy group. CONCLUSION: CO(2) pneumoperitoneum resulted in severe peritoneal acidosis that was unaltered by heating and humidification with or without bicarbonate. Alteration in peritoneal pH may conceivably be responsible for providing an environment favorable for tumor-cell implantation during laparoscopy. PMID- 15791378 TI - Effectiveness of esophageal manometry in predicting the outcome of children with primary GER after floppy Nissen-Rossetti wrap. AB - BACKGROUND: Surgical treatment of gastroesophageal reflux (GER) can result in many postoperative problems because of an incorrect indication or an unsuitable fundoplication. Many preoperative tests have been suggested to perform a "tailored fundoplication," but there is no clear evidence as to which is the best. The aim of our study was to define the effectiveness of esophageal manometry in predicting the outcome of children who need fundoplication because of refractory primary gastroesophageal reflux. METHODS: Thirty-two children were included in the study. Patients with gastroenterologic and respiratory symptoms numbered 10 (31%) and 22 (69%), respectively. The preoperative motility pattern was (1) inappropriate relaxations alone in nine patients (28%) and associated with esophageal body dysmotility in one patients (3%); (2) low-pressure lower esophageal sphincter (LES), alone in 13 patients (41%) and associated with body dysmotility in eight (25%). Motility pattern of the esophageal body was abnormal in nine children. Manometric anomalies were absent only in one case. RESULTS: Low pressure of the LES associated with atypical esophageal motility disorders was significantly higher in the gastroenterological group and in the older patients. Low pressure of the LES associated with good esophageal activity was significantly higher in the respiratory group and in the patients younger than 24 months. There was no correlation between motility pattern and outcome, and no statistically significant differences were found between pressure values at the different levels and symptoms, outcome, and age groups. CONCLUSIONS: Esophageal manometry is not mandatory to predict the outcome of patients undergoing laparoscopic Nissen-Rossetti fundoplication for refractory primary GER treatment. PMID- 15791379 TI - The Fenyo-Lindberg scoring system for appendicitis increases positive predictive value in fertile women--a prospective study in 455 patients randomized to either laparoscopic or open appendectomy. AB - BACKGROUND: Suspected appendicitis is one of the most common indications for acute laparotomy or laparoscopy. The negative laparotomy and laparoscopy rates are high, often in the range of 15-30%, and especially high in some groups of patients such as women of child-bearing age and young patients. Different scoring systems have been introduced in order to improve diagnostic accuracy. The aim of the present study was to analyse the outcome of the Fenyo-Lindberg scoring system in a prospectively randomized multicenter trial and to analyze how well the score performed in stratified subgroups. METHODS: The variables of the Fenyo-Lindberg scoring system were collected in a prospective study comparing laparoscopic and open surgery in suspected appendicitis and with four participating centers. None of the hospitals had used the scoring system previously. Since surgeons were unfamiliar with the score, they could not use it as a diagnostic aid. When comparing the score with the clinical outcome, retrospectively, the investigators interpreting the score were blinded regarding the surgical outcome. RESULTS: Positive predictive value (PPV) of the Fenyo-Lindberg score was higher than that of the surgeon's clinical diagnosis in the patient cohort [0.90 vs 0.79 (p < 0.001)]. The score demonstrated an improvement of PPV in women [0.83 vs 0.70 (p < 0.01)]. PPV was increased in women between 15 and 50 years of age. In women aged 15-30 years and 31-50 years PPV increased from 0.69 to 0.82 and 0.68 to 0.86, respectively (p < 0.01). Both the sensitivity (0.77) and the specificity (0.69) of the score were, however, low. CONCLUSION: The Fenyo-Lindberg score is an inexpensive clinical tool that may improve the diagnostic accuracy for acute appendicitis in women of childbearing age, which is a group of patients where the diagnostic accuracy usually is low and where the arsenal of diagnostic tools such as computed tomography is limited because of radiation. The low specificity of the score in women of childbearing age must, however, be kept in mind. PMID- 15791380 TI - Learning curves and impact of psychomotor training on performance in simulated colonoscopy: a randomized trial using a virtual reality endoscopy trainer. AB - BACKGROUND: The aim of this study was to analyze the learning curve for the GI Mentor II endoscope trainer and to determine whether psychomotor training can contribute to an improvement in the performance of virtual colonoscopy. METHODS: To analyze the learning curve, 28 subjects were divided into three groups on the basis of their experience with gastrointestinal (GI) endoscopy: experienced surgeons (group 1, performed > 200 endoscopic procedures, (n = 8)) residents (group 2, performed < 50 endoscopic procedures, (n = 10)); and medical students (group 3, never performed GI endoscopy, (n = 10)). The participants were tested on the GI Mentor II virtual reality simulator 10 consecutive times. Assessment of the learning curve was based on the following three parameters: time used, number of punctured balloons, and number of wall collisions. In the second part of the study, 20 subjects who had never performed GI endoscopy were included. After performing a virtual colonoscopy, they were randomized to a group that received psychomotor training and a control group. Finally, all subjects performed a virtual colonoscopy. Assessment of endoscopic skills during the colonoscopy was based on nine parameters dealing with psychomotor skills. RESULTS: The learning curve for time expended reached a plateau after the second repetition for group 1 (Friedman's test, p < 0.05), after the fifth repetition for group 2 (p < 0.05), and after the seventh repetition for group 3 (p < 0.05). Experienced surgeons did not improve their scores for regarding number of balloons punctured or number of wall collisions (p > 0.05), indicating the absence of a learning curve for these parameters. Group 2 improved their scores up to the fourth and fifth repetitions, respectively (p < 0.05), and group 3 up to the fifth and seventh repetitions, respectively (p < 0.05). Experienced surgeons achieved the best performance, followed by group 2 and then group 3. The surgeons who had received psychomotor training performed the second virtual colonoscopy significantly faster than the control group (Mann-Whitney test, p < 0.001) and made significantly greater improvement in all other parameters as well. CONCLUSIONS: There were different learning curves for surgeons depending on their endoscopic background. The familiarization rate on the simulator was proportional to the endoscopic experience of the surgeons. Psychomotor training had a significant effect on the performance of a simulated colonoscopy. PMID- 15791381 TI - The impact of introducing laparoscopic donor nephrectomy to an established renal transplant program. AB - BACKGROUND: Although the advent of hand-assisted laparoscopic donor nephrectomy (HLDN) has had a positive impact on the donor pool, there is still some concern about its safety. The aim of this study was to assess the impact of a change in surgical access to live-donor nephrectomy on donor-related complication rates, the renal function of the donor, and the graft function of the recipient. METHODS: At our hospital, HLDN was introduced in 1998. Thereafter, we compared 49 consecutive donors undergoing open donor nephrectomy (ODN) between 1987 and 2002 with 57 consecutive donors undergoing HLDN between 1998 and 2002. Donor renal and recipient graft functions were assessed by measuring creatinine levels and urine output, with the addition of warm and cold ischemia time and dialysis requirements in the latter group. Data are presented as means (+/-SD) and analyzed with the Student t-test or Fisher's exact test. RESULTS: The ODN and HLDN donors were comparable for age, gender, body mass index, renovascular anatomy, and preoperative creatinine. Estimated blood loss (370 +/- 280 vs 168 +/ 160 ml, p < 0.0001), time to resumption of oral intake (1.7 +/- 0.5 vs 1.3 +/- 0.7 days, p = 0.01), duration of intravenous narcotic requirements (23 +/- 0.7 vs 1.7 +/- 1.0 days, p < 0.0001), and hospital stay (4.2 +/- 1.4 vs 2.9 +/- 1.3 days, p < 0.0001) were significantly decreased after HLDN. There were no significant differences between ODN and HLDN in operating time (204 +/- 46 vs 202 +/- 49 min), donor-related complication rates (12.2% vs 14%), or donor renal and recipient graft functions. CONCLUSION: The introduction of HLDN to an established renal transplant program led to an improved short-term outcome without any increase in donor-related complication rates or delay in recipient graft function. PMID- 15791382 TI - The Italian Group for LAP-BAND: predictive value of initial body mass index for weight loss after 5 years of follow-up. AB - BACKGROUND: Laparoscopic of the LAP-BAND System placement stage of obesity is a safe operation, but its indication in terms of stage of obesity is controversial. The aim of this study was to evaluate the 5 years stage of obesity results for weight loss in patients with varying preoperative ranges of body mass index (BMI). METHODS: Data were obtained from the Italian Collaborative Study Group for LAP-BAND System (GILB) registry. Detailed information was collected on a specifically created database (MS Access 2000) for patients operated on in Italy from January 1996 to 2003. Patients operated on between January 1996 and December 1997 were allocated to four groups according to preoperative BMI range: 30-39.9 kg/m(2) (group A), 40-49.9 kg/m(2) (group B), 50-59.9 kg/m(2) (group C), and =60 kg/m(2) (group D) percent estimated weight loss respectively. Postoperative complications, mortality, BMI, BMI loss, and (%EWL) were considered in each group. Data are expressed as mean +/- SD, except as otherwise indicated. Statistical analysis was done by means of Fisher's exact test, and p < 0.05 was considered significant. RESULTS: After 5 years from LAP-BAND System surgery, 573 of 3,562 patients were eligible for the study. One hundred fifty-five of 573 (27.0%) were lost to follow-up, 24 of 418 (5.7%) underwent band removal due to complications (gastric pouch dilation, band erosion), eight of 418 (1.9%) were converted to other bariatric procedures, five of 418 (1.2%) died of causes not related to the operation or the band, and 381 of 573 (66.5%) were available for follow-up. Based on 96, 214, 64, and seven patients their preoperative BMI, Were allocated to groups A, B, C, and D, respectively. At time of follow-up mean BMI was 27.5 +/- 5.2 in group A, 31.6 +/- 4.7 in group B, 37.6 +/- 17.3 in group C, and 41.4 +/- 6.9 kg/m(2) in group D. Mean BMI loss was 9.8 +/- 5.4, 12.9 +/- 5.2, 15.8 +/- 8.1, and 23.2 +/- 4.9 kg/m(2), respectively, in groups A, B, C, and D. Mean %EWL at the same time was 54.6 +/- 32.3 in group A, 54.1 +/- 17.2 in group B, 51.6 +/- 35 in group C, and 59.l +/- 17.1 in group D. CONCLUSION: Initial BMI in this series did not correlate with %EWL 5 years after the operation. In fact %EWL was almost the same in each group, independent of preoperative weight. Initial BMI was an accurate indicator of the results obtained 5 years after LAP BAND in group C (50-59.9 kg/m(2)) and D (=60 kg/m(2)) patients, who remained morbidly obese despite their %EWL. PMID- 15791383 TI - Effects of the aldosterone receptor antagonist potassium canrenoate on renal blood flow and urinary output during prolonged increased intraabdominal pressure (IAP) in pigs. AB - BACKGROUND: Increased intraabdominal pressure can be found after major abdominal trauma and necrotizing pancreatitis and is used during laparoscopic surgery. The purpose of this study was to investigate the effect of the aldosterone receptor antagonist (potassium canrenoate) on renal hemodynamics and urinary output in pigs during increased intraabdominal pressure (IAP). METHODS: The IAP was kept at 30 mmHg for 3 h by instillation of Ringer's solution into the peritoneal cavity. Eight animals were treated with potassium canrenoate and eight animals served as controls. Renal blood flow, hormones in femoral artery blood, and the urinary output were measured. RESULTS: The administration of potassium canrenoate was followed by increased aldosterone concentrations in arterial blood, increased blood concentration of potassium, and increased concentration of sodium in the urine, indicating satisfactory inhibition of aldosterone. Potassium canrenoate did not cause changes in cardiac output and arterial pressure. It did not affect the renal vascular resistance that increased at an IAP of 30 mmHg, or the renal blood flow that remained constant during the experiments. The group treated with potassium canrenoate had higher mean urinary output than the controls, but the difference was not significant. CONCLUSION: Increased IAP in pigs is associated with markedly reduced urinary output and increased serum concentrations of aldosterone. Although the urinary output did not increase significantly, the increased sodium concentration in the urine of canrenoate-treated animals suggests that the high blood level of aldosterone contributes to the oliguria under increased IAP. PMID- 15791384 TI - Video-assisted transcervical thymectomy. AB - BACKGROUND: The optimal surgical approach for complete removal of the thymus gland has long been debated. In this report, the excision of the entire gland through a transcervical incision using video-assisted techniques is described. METHODS: Ten patients, including one with thymoma and myasthenia gravis, underwent surgery via the transcervical approach. After standard dissection up to the level of the innominate vein and ligation of the thymic vessels, a laparoscope was inserted into the mediastinum. In the patient with thymoma, the operation was completed by a small incision in the third intercostal space. RESULTS: No perioperative mortality or long-term morbitity was observed. The mean hospital stay was 69.6 h. After a mean follow-up period of 63.8 months, eight patients displayed complete remission, whereas one continued to receive minimal medication. The patient with thymoma showed considerable improvement, but remained on same medical regimen No complications were seen throughout the study. CONCLUSION: Video-assisted thymectomy improves effectiveness of the transcervical approach for thymectomy with a minimum of trauma and excellent results. PMID- 15791386 TI - Laparoscopic adrenalectomy for Conn's syndrome complicated by ipsilateral congenital pelvic kidney. AB - A patient presented with hypertension, hypokalemia, and a 1.2-cm left adrenal tumor. Conn's syndrome was diagnosed, for which laparoscopic adrenalectomy is now the therapy of choice. This case was complicated by an ipsilateral ectopic pelvic kidney. A laparoscopic left adrenalectomy was performed via a lateral transabdominal approach. Without the usual anatomic landmark of the ipsilateral kidney, the left adrenal gland was difficult to identify, so intraoperative ultrasound was used to locate the lesion. Postoperatively, the patient's blood pressure and potassium normalized. This is the first documented report of a laparoscopic adrenalectomy performed for adrenal adenoma with the anatomic disruption of an ipsilateral pelvic kidney. PMID- 15791385 TI - Hemorrhagic adrenal pseudocyst: laparoscopic treatment. AB - BACKGROUND: The incidence of adrenal hemorrhagic pseudocyst is very low. A total of 613 adrenal cysts and 85 hemorrhagic pseudocysts have been reported. A laparoscopically diagnosed and resolved case is presented here, together with the current diagnostic and therapeutic procedures. METHODS: A 40-year-old woman was admitted because of an asymptomatic nonfunctional right adrenal tumor. Right laparoscopic adrenalectomy was performed, 8-cm cyst which found an with thick walls and organized hematic content. The postoperative course was uneventful. Follow-up was 14 months. The pathology was an adrenal hemorrhagic pseudocyst. DISCUSSION: A total of 56% of adrenal cysts are pseudocysts. One third of them have hematic content. They may present as an asymptomatic finding with nonspecific symptoms or as a hormone secreting or complicated tumor. Their vascular etiology is not yet totally accepted. There is a tendency for intracystic bleeding. it is advisable to evaluate the hormonal profile and morphologic characteristics in all cases. Treatment options include needle aspiration, percutaneous drainage, and cyst or gland resection. Laparoscopic excision should be evaluated. PMID- 15791387 TI - Dual endoscopically assisted endoluminal colostomy reversal. PMID- 15791388 TI - A cost-utility analysis of treatment options for inguinal hernia in 1,513,008 adult patients. PMID- 15791389 TI - Isolated bilateral abducent nerve palsy due to a spontaneous left-side dural carotid cavernous fistula Type Barrow C. PMID- 15791390 TI - Risk factors for critical illness polyneuromyopathy. AB - Although numerous clinical, laboratory, and pharmacological variables have been reported as significant risk factors for critical illness polyneuromyopathy (CIPM), there is still no consensus on the aetiology of this condition. Objectives of the study were to assess the clinical and electrophysiological incidence and risk factors for CIPM.A cohort of critically ill patients was observed prospectively for a one-month period and the association between neuromuscular involvement and various potential risk factors was evaluated. Sixty one critically ill patients completed the follow-up (30 women, 31 men, median age 59 years).CIPM development was detected clinically in 17 patients (27.9 %) and electrophysiologically in 35 patients (57.4 %). CIPM was significantly associated with the presence and duration of systemic inflammatory response syndrome and the severity of multiple, respiratory, central nervous, and cardiovascular organ failures. The median duration of mechanical ventilation was significantly longer in patients with CIPM than in those without (16 vs 3 days, p<0.001). Independent predictors of CIPM obtainable within the 1(st) week of critical illness were the admission sequential organ failure assessment score (odds ratio [OR], 1.15; 95% confidence interval [CI], 1.02-1.36), the 1(st) week total sequential organ failure assessment scores (OR, 1.14; 95 % CI, 1.06-1.46) and the 1(st) week duration of systemic inflammatory response syndrome (OR, 1.05; 95% CI, 1.01 1.15). They were able to correctly predict the development of CIPM at the end of the 1(st) week in about 80% of critically ill cases.In conclusion, the presence and duration of systemic inflammatory response syndrome and the severity of multiple and several organ failures are associated with increased risk of the development of CIPM. PMID- 15791391 TI - Microsatellite markers reveal high allelic variation in natural populations of Cryptomeria japonica near refugial areas of the last glacial period. AB - Using 11 microsatellite markers, we investigated the allelic variation and genetic structure of Cryptomeria japonica, across most of its natural distribution. The markers displayed high levels of polymorphism (average gene diversity=0.77, average number of alleles=24.0), in sharp contrast to the lower levels of polymorphism found in allozyme and cleaved amplified polymorphic sequence markers in previous studies. Little genetic differentiation was found among populations (F(ST)=0.028, P<0.001), probably because the species is wind pollinated and long-lived. No clear relationship between Nei's genetic distances and geographical locations of the populations were found using the principal coordinate and unweighted pair-group method with arithmetic averaging analyses. The lack of such trends might be due partly to microsatellite homoplasy arising from mutation blurring the genealogical record. However, there was a trend towards high allelic diversity in five populations (Ashitaka, Ashiu, Oki-Island, Yakushima-Island-1 and -2), which are very close to, or in, refugial areas of the last glacial period as defined by Tsukada based on pollen analysis data and current climatic divisions. We postulate that these refugial populations might have been less affected by genetic drift than the other populations due to their relatively large size. PMID- 15791392 TI - Effects of hypoxic and osmotic stress on the free D-aspartate level in the muscle of blood shell Scapharca broughtonii. AB - Blood shell, Scapharca broughtonii, contains large quantities of free D-aspartate comparable to free L-aspartate in its tissues. When the shell was reared in hypoxic seawater, D-aspartate as well as L-aspartate in the foot muscle decreased rapidly, and their total level became about one-fourth within 24 hr. None of the other amino acids examined showed a similar behavior, but many of them rather increased during the same period. The increase in L-alanine was especially remarkable and was almost equal to the sum of the decrease in aspartate enantiomers. When the shell that had been acclimated to hypoxic seawater for 96 hr was transferred to normoxic seawater, all the amino acid levels mostly returned to the control levels within 96 hr. In contrast to these effects of hypoxic stress, hyperosmotic stress of 150% seawater had no effect on the D- and L-aspartate levels in the same tissue. These results suggest that D-aspartate is involved in anaerobic energy metabolism of this bivalve as well as L-aspartate, whose vital role in anoxia-tolerant bivalves is well known. PMID- 15791393 TI - Repeated-testing of place preference expression for evaluation of anti-craving drug effects. AB - In addiction research, the conditioned place preference (CPP) paradigm is a widely used animal model of conditioned reward. Usually, CPP development is studied, while only few studies examine CPP expression. In the present study, the suitability of a schedule allowing repeated testing of CPP expression was evaluated. Two groups of rats were either conditioned with cocaine or morphine then the repeated-testing-schedule was applied. This schedule consisted of four repeated applications of a sequence of drug- (i.e. cocaine or morphine), saline- and anti-craving-drug- (i.e. acamprosate, naloxone, their joint administration or saline as internal control) tests. Methodologically, the repeated-testing schedule produced stable CPP expression in both groups over 12 subsequent tests. In conclusion, it is suggested as a useful method to study effects of anti craving-drugs on CPP expression, thereby reducing the overall number of experimental animals. The evaluation of the anti-craving-drug effects revealed that neither acamprosate and naloxone given separately nor their combined administration significantly reduced cocaine- or morphine-CPP expression. Thus, we suggest that these anti-craving-drugs are unlikely to be effective for relapse prevention in cocaine- or morphine-addicts. PMID- 15791394 TI - Synthesis and utilization of 13C and 15N backbone-labeled proline: NMR study of synthesized oxytocin with backbone-labeled C-terminal tripeptide amide. AB - The 13C and 15N backbone-labeled proline was prepared using Oppolzer's method based on application of a sultam as chiral auxiliary. This isotopomer was used in the synthesis of the 13C, 15N backbone-labeled C-terminal tripeptide amide fragment of neurohypophyseal hormone oxytocin. Finally, this tripeptide amide was coupled by segment condensation with N-Boc- or N-Fmoc-tocinoic acid, followed by N-deprotection with TFA or piperidine. The labeled oxytocin exhibited biological activity identical with that of natural oxytocin. A detailed 1H, 13C and 15N NMR study confirmed the assigned oxytocin conformation containing a beta-turn in the cyclic part of the molecule, stabilized by H-bond(s) that can be perturbed by the C-terminal tripeptide amide moiety as indicated by comparison of NMR data for both the tocine ring in oxytocin and tocinoic acid. PMID- 15791395 TI - Expression profiling using human tissues in combination with RNA amplification and microarray analysis: assessment of Langerhans cell histiocytosis. AB - Advances in molecular genetics have led to sequencing of the human genome, and expression data is becoming available for many diverse tissues throughout the body, allowing for exciting hypothesis testing of critical concepts such as development, differentiation, homeostasis, and ultimately, disease pathogenesis. At present, an optimal methodology to assess gene expression is to evaluate single cells, either identified physiologically in living preparations, or by immunocytochemical or histochemical procedures in fixed cells in vitro or in vivo. Unfortunately, the quantity of RNA harvested from a single cell is not sufficient for standard RNA extraction methods. Therefore, exponential polymerase chain reaction (PCR) based analyses, and linear RNA amplification including amplified antisense (aRNA) RNA amplification and a newly developed terminal continuation (TC) RNA amplification methodology have been used in combination with microdissection procedures such as laser capture microdissection (LCM) to enable the use of microarray platforms within individual populations of cells obtained from a variety of human tissue sources such as biopsy-derived samples {including Langerhans cell histiocytosis (LCH)} as well as postmortem brain samples for high throughput expression profiling and related downstream genetic analyses. PMID- 15791396 TI - Preparation of new amino acid complex nanoparticles of bismuth and leucine. AB - The new amino acid complex nanoparticles of bismuth and leucine can be prepared very easily by a room temperature solid-solid reaction. The various characterizations indicate that the formula of the complex is BiCl[(CH(3))(2)CHCH(2)CHNH(2)COO](2)1.5H(2)O. The crystal structure of the solid complex belongs to monoclinic system with the lattice parameters: a = 1.6036 nm, b = 1.9903 nm, c = 2.1979 nm and beta=108.06 degrees . The new solid complex is nanoparticles with average size about 80 nm. PMID- 15791397 TI - Paragonimus skrjabini Chen, 1959 (Digenea: Paragonimidae) and related species in eastern Asia: a combined molecular and morphological approach to identification and taxonomy. AB - A molecular and morphometric investigation is reported on the species complex of mammalian lungflukes of which Paragonimus skrjabini Chen, 1959 and P. miyazakii Kamo, Nishida, Hatsushika & Tomimura, 1961 are the best-known examples. This species complex (here called the P. skrjabini complex) is shown to be monophyletic using DNA sequences from the nuclear ITS2 region and the mitochondrial cox1 gene. The latter marker permits the discrimination of populations, some previously named as distinct species, from various geographical locations in China and Japan. Morphometric analysis of a number of variables accords remarkably closely with the molecular results. Main findings are that (1) nominal P.skrjabini from Fujian Province in eastern China is phylogenetically very close to P. miyazakii from Japan. It is proposed that both taxa should be referred to the same subspecies as P. krjabini miyazakii. (2) Populations from Guangdong Province, Guangxi Zhuang Autonomous Region, Sichuan and Hubei should be referred to P. skrjabini skrjabini. Specimens of P. skrjabini from the Yangtze basin in Sichuan and Hubei are remarkably similar to one another according to genetic and morphometric data, but are not so different from Guangdong populations that they should be referred to a different subspecies at this stage. Specimens of P. skrjabini from Yunnan have not been assigned to a subspecies at this stage. (3) Partly following earlier studies, we regard the following as synonyms of P. skrjabini: P. miyazakii (reduced to subspecific status); P. szechuanensis Chung & Tsao, 1962 (probably belongs to P. s. skrjabini); and P. hueitungensis Chung, Hsu, Ho, Kao, Shao, Chiu, Pi, Liu, Ouyang, Shen, Yi & Yao, 1975 (probably belongs to P.s.skrjabini ); P. veocularis (Chen & Li, 1979) (specimens from Fujian regarded as belonging to this species should be referred to P.skrjabini miyazakii: the subspecific status of specimens from the type locality in northern Sichuan is unclear). A number of questions remain unresolved. The name P. hokuoensis Ho & Chung, 1964 was proposed for two individual metacercariae of distinctive appearance from southern Yunnan. DNA sequences from very similar metacercariae from the same locality place this nominal species within, or sister to, the P. skrjabini complex. As yet, nothing is known regarding adult morphology or biology of this taxon and we retain it here as a distinct species. P. heterorchis (Zhou, Pang & Hsiang, 1982) might be a synonym of P. skrjabini: the form of the metacercaria provides evidence against this view and further work is required. P. macrorchis Chen, 1962 has probably been confused with P. skrjabiniin China. Within China, the former probably occurs only on Hainan Island, although P. fukienensis Tang & Tang, 1962 from Fujian Province could be a synonym. PMID- 15791398 TI - A new entomopathogenic nematode, Steinernema robustispiculum n. sp. (Rhabditida: Steinernematidae), from Chumomray National Park in Vietnam. AB - Steinernema robustispiculum n. sp. (Rhabditida: Steinernematidae) was isolated from woodland in Chumomray National Park, Sason, Sathay, Kontum, Vietnam. Its morphology, morphometrics, cross-hybridisation and the ITS-rDNA sequence analysis revealed that S. robustispiculum clearly differs from other known Steinernema spp. As in the cases of S. intermedium (Poinar, 1985), S. robustispiculum has very robust spicules, but it can be distinguished by the longer tail of the infective juvenile, lower E%, shorter spicules, the shape of the spicules, the number of genital papillae in the caudal region and the presence of a mucron on the male tail. S. robustispiculum has a lateral field resembling that of S. sangi Phan, Nguyen & Moens, 2001, but can be distinguished by a higher E%, higher D%, smaller length to width ratio of the spicules and the morphology of both the spicule head (manubrium) and the dorsal lobe of the spicule. The morphometrics of infective juveniles of S. robustispiculum are similar to those of S. monticolum Stock, Choo & Kaya, 1997; these species can be distingusihed by the position of the excretory pore, the smaller length to width ratio of the spicules, and the length and morphology of the spicule head (manubrium). The phylogenetic relationships within Steinernema Travassos, 1927, including the newly sequenced Vietnamese species S. robustispiculum n. sp., S. loci Phan, Nguyen & Moens, 2001, S. thanhi Phan, Nguyen & Moens, 2001 and S. sangi, are presented based on analyses of the ITS-rDNA. The ITS RFLP profiles obtained from 17 different restriction enzymes are also presented. PMID- 15791399 TI - First report of a bucephalid digenean from an apogonid teleost: Prosorhynchoides apogonis n. sp. from Cheilodipterus macrodon on the southern Great Barrier Reef, Australia. AB - Prosorhynchoides apogonis n. sp. (Digenea: Bucephalidae) is described from the intestine of the apogonid Cheilodipterus macrodon on the southern Great Barrier Reef, Australia. The new species is differentiated from other species of Prosorhynchoides Dollfus, 1929 by the configuration of its digestive system, shape and distribution of its vitelline follicles, and the shape and extent of its uterus. This is the first bucephalid to be described from the teleost family Apogonidae, and our records suggest that this species is strongly host-specific, at least to the genus Cheilodipterus. The host family is consistent with the pattern of Prosorhynchoides being reported from a very wide range of piscivorous fish families. As a result of comparisons made in the description of this species, 18 species formerly included in Bucephaloides Hopkins, 1954,Bucephalopsis Diesing, 1855 and Neobucephalopsis Dayal, 1948 are transferred to Prosorhynchoides, 16 as new combinations and two as replacement names to prevent secondary homonymy. PMID- 15791400 TI - Prodajus curviabdominalis n. sp. (Isopoda: Epicaridea: Dajidae), an ectoparasite of mysids, with notes on morphological changes, behaviour and life-cycle. AB - Adults and larvae of a new ectoparasitic isopod, Prodajus curviabdominalis n. sp., are described from the mysid Siriella okadai Ii collected from the Seto Inland Sea, western Japan. The adult female is found within the host marsupium with the cephalon directed posteriorly, whereas the dwarf adult male attaches to the ventral surface of the female pleon. The cryptoniscid larva usually attaches to the second or third abdominal somite of the host, using an oral sucker. Mature adults of the new species are distinguished from all other congeners by: pleon of ovigerous female strongly curved dorsally, with large swellings on ventral side; pereon of ovigerous female narrow; exopods on male uropods present; male pleon short and thick. This is the third record of a member of the Dajidae from Japan. The behaviour of the cryptoniscid larvae of the new species on the host mysid was also observed using a video camera. Larvae moved from the first attachment site, usually the second or third abdominal somite of the host mysid, into the marsupium. When host oostegites were not fully developed, larvae entered beneath the host carapace until her marsupium was fully formed. The host infected by a female P. curviabdominalis moved the oostegites rhythmically, an action which may aid the respiration of the parasite. PMID- 15791401 TI - Clestobothrium gibsoni n. sp. (Cestoda: Bothriocephalidae) from the bullseye grenadier Bathygadus macrops Goode & Bean (Macrouridae) in the Gulf of Mexico. AB - Clestobothrium gibsoni n. sp. is described from the intestine of the bullseye grenadier Bathygadus macrops (Macrouridae) collected in the northeastern Gulf of Mexico. The new species can be distinguished from both C. crassiceps (Rudolphi, 1819), the type-species of the genus, and C. neglectum (Lonnberg, 1893), the second species in the genus, by its unique ovary, which is transversely elongate and tapered on both ends rather than being U-or H-shaped, and by having more testes (60-65 vs 40-50 and 35-45 per proglottid). The new species is also compared to Bothriocephalus acheilognathi Yamaguti, 1934 and B. kivuensis Baer & Fain, 1954, two species with similar scoleces which have previously been assigned to Clestobothrium . The generic diagnosis of Clestobothrium is amended to include species with either anoperculate or operculate eggs. This is the first report of an adult tapeworm from a species of Bathygadus , and the first report of a species of Clestobothrium Luhe, 1899 (Bothriocephalidae) from the Gulf of Mexico. PMID- 15791402 TI - Aggregata andresi n. sp. (Apicomplexa: Aggregatidae) from the ommastrephid squid Martialia hyadesi in the SW Atlantic Ocean and some general remarks on Aggregata spp. in cephalopod hosts. AB - A new species of coccidian, Aggregata andresi, is described from the digestive tract of the flying squid Martialia hyadesi, an ommastrephid squid that lives in cold subantarctic waters in the Southwest Atlantic. Gamogonic and sporogonic stages were observed in the digestive tract of 96.5% studied hosts. Oocysts were ovoid to subspheroid in shape, 170-590 microm in length and 200-530 microm in width. Each oocyst contained approximately 45,000 sporocysts. Sporocysts measured 9.5-10 microm in length and 8-8.5 microm in width. The surface of the sporocyst wall was smooth and the cyst wall thick. Each sporocyst contained three sporozoites measuring 16-20 microm in length and 2-2.5 microm width. A. andresi is the second species of Aggregata to be reported from a nerito-oceanic cephalopod host. PMID- 15791403 TI - Redescription of Argulus multipocula Barnard, 1955 (Crustacea: Branchiura) collected on the west coast of South Africa. AB - Argulus multipocula Barnard, 1955 was originally described from a single female found in a littoral sample from the east coast of South Africa. We present a redescription of this species based on light and scanning electron microscope studies of 25 females collected from the southern mullet Liza richardsonii (Smith) on the west coast of South Africa. PMID- 15791407 TI - Nuclear architecture - on higher ground. A report on the Third Elmau Conference on Nuclear Organization: from Basic Science to Application. PMID- 15791408 TI - New types of mouse centromeric satellite DNAs. AB - Genomic databases do not contain complete sequences of the centromeric regions. We created a pUC19-based library of DNA fragments from isolated chromocentres of interphase nuclei. In this library we have found major satellite (MaSat) and two new satellite sequences - MS3 and MS4. The computer analysis of MS3 and MS4 sequences by alignment, fragment curved state and search for MAR motifs in comparison with the mouse major and minor satellite (MiSat) DNA has shown them to be new satellite fragments. Southern blot of MS3 and MS4 with total DNA digested by restriction enzymes shows the ladder characteristic of satellite DNA. 2.2% of the total DNA consists of MS3, the monomer of which is 150 bp long. The MS4 monomer is 300 bp long and accounts for 1.6% of the total DNA. On metaphase chromosomes MS3 and MS4 are located at the centromeric region. FISH analysis of L929 nuclei during the cell cycle showed relative positions of MaSat, MiSat, MS3, and MS4. All mapped satDNA fragments except MaSat belong to the outer layer of the chromocentres in the G0/G1 phase. MS3 is likely to be involved in the centromere formation. The mouse genome contains at least four satDNA types: AT rich (MaSat and MiSat), and CG-rich (MS3 and MS4). PMID- 15791409 TI - Polymerase chain reaction-based suppression of repetitive sequences in whole chromosome painting probes for FISH. AB - We have developed a method to suppress the PCR amplification of repetitive sequences in whole chromosome painting probes by adding Cot-1 DNA to the amplification mixture. The repetitive sequences in the Cot-1 DNA bind to their homologous sequences in the probe library, prevent the binding of primers, and interfere with extension of the probe sequences, greatly decreasing PCR efficiency selectively across these blocked regions. A second labelling reaction is then done and this product is resuspended in FISH hybridization mixture without further addition of blocking DNA. The hybridization produces little if any non-specific binding on any other chromosomes. We have been able to successfully use this procedure with both human and rat chromosome probes. This technique should be applicable in producing probes for CGH, M-FISH and SKY, as well as reducing the presence of repetitive DNA in genomic libraries. PMID- 15791410 TI - Molecular and cytogenetic characterization of site-specific repetitive DNA sequences in the Chinese soft-shelled turtle (Pelodiscus sinensis, Trionychidae). AB - A novel family of repetitive DNA sequences that are components of constitutive heterochromatin were cloned from BglI-digested genomic DNA of the Chinese soft shelled turtle (Pelodiscus sinensis, Trionychidae), and characterized by filter hybridization and chromosome in-situ hybridization. The BglI-family of repetitive sequences were classified into four types by their genome organization and chromosomal distribution as follows: the repeated sequences located on (1) two pairs of microchromosomes, (2) four pairs of microchromosomes,(3) about half the number of microchromosomes and (4) the interstitial region of the short arm of chromosome 2. The presence of microchromosome-specific repetitive sequences has also been reported in the Struthioniformes and Galliformes, suggesting that turtle chromosomes retain some similarity to the chromosome structure as well as the karyotypes of avian species. PMID- 15791411 TI - Chromosomal polymorphism and comparative painting analysis in the zebra finch. AB - The zebra finch (Taeniopygia guttata) is often studied because of its interesting behaviour and neurobiology. Genetic information on this species has been lacking, making analysis of informative mutants difficult. Here we report on an improved cytological method for preparation of metaphase chromosomes suitable for fluorescent in situ hybridization of adult birds. We found that individual chicken chromosome paints usually hybridized to single zebra finch chromosomes, indicating only minor chromosomal rearrangements since the evolutionary divergence of these two species, and suggesting that the genomic location of chicken genes will predict the location of zebra finch orthologues. Chicken chromosome 1 appears to have split into two macrochromosomes in zebra finches, and chicken chromosome 4 paint hybridizes to a zebra finch macrochromosome and a microchromosome. This pattern was confirmed by mapping the androgen receptor (AR), which is located on chicken chromosome 4 but on a zebra finch microchromosome. We detected a telocentric/submetacentric polymorphism of chromosome 6 in our colony of zebra finches, and found that the polymorphism was inherited in a Mendelian pattern. PMID- 15791412 TI - Plasticity of histone modifications across the invertebrate to vertebrate transition: histone H3 lysine 4 trimethylation in heterochromatin. AB - Histone posttranslational modifications mediate establishment of structurally and functionally distinct chromatin compartments of eukaryotic nuclei. The association of different histone modifications with euchromatic and heterochromatic compartments is relatively conserved in highly divergent model organisms such as Drosophila and mammals. However, some differences between these model systems have been uncovered while limited data are available from organisms nearer the invertebrate-vertebrate transition. We identified a chromatin compartment in both diploid and endocycling cells of the urochordate, Oikopleura dioica, enriched in heterochromatic histone modifications and DNA methylation. Surprisingly, this compartment also contained high levels of histone H3 trimethylated at lysine 4 (H3 Me(3)K4), a modification thus far associated with actively transcribed sequences. Although in Drosophila and mouse cells, H3 Me(3)K4 was prevalently associated with euchromatin, we also detected it in their pericentromeric heterochromatin. We further showed that H3 Me(3)K4 abundance was not necessarily proportional to local levels of transcriptional activity in either euchromatin or heterochromatin. Our data indicate greater plasticity across evolution in the association of histone lysine methylation with functionally distinct chromatin domains than previously thought and suggest that H3 Me(3)K4 participates in additional processes beyond marking transcriptionally active chromatin. PMID- 15791413 TI - Comparison of Dam tagging and chromatin immunoprecipitation as tools for the identification of the binding sites for S. pombe CENP-C. AB - We have established the identity of the Schizosaccharomyces pombe homologue of vertebrate CENP-C and Saccharomyces cerevisiae MIF2p and have used it to compare Dam tagging and chromatin immunoprecipitation (ChiP)as tools for the mapping of protein binding sites on DNA. ChiP shows that S. pombe CENP-C binds to the central core and inner repeats of the S. pombe centromere. It binds weakly, however, to the outer repeats. The binding pattern is thus similar to that of S. pombe CENP-A. Dam-tagged S. pombe CENP-C, however, methylates the entire centromere and 5 kb of flanking DNA. This comparison suggests that Dam tagging is less precise as a tool for mapping DNA binding sites than ChiP. We have also used the Dam tagging technique to address the question of whether there is any CENP-C binding to the ribosomal DNA in S. pombe and find none. PMID- 15791414 TI - Mapping genomic rearrangements in titi monkeys by chromosome flow sorting and multidirectional in-situ hybridization. AB - We developed chromosome painting probes for Callicebus pallescens from flow sorted chromosomes and used multidirectional chromosome painting to investigate the genomic rearrangements in C. cupreus and C. pallescens. Multidirectional painting provides information about chromosomal homologies at the subchromosomal level and rearrangement break points, allowing chromosomes to be used as cladistic markers. Chromosome paints of C. pallescens were hybridized to human metaphases and 43 signals were detected. Then, both human and C. pallescens probes were hybridized to the chromosomes of another titi monkey, C. cupreus. The human chromosome paints detected 45 segments in the haploid karyotype of C. cupreus. We found that all the syntenic associations proposed for the ancestral platyrrhine karyotype are present in C. cupreus and in C. pallescens. The rearrangements differentiating C. pallescens from C. cupreus re one inversion, one fission and three fusions (two tandem and one Robertsonian)that occurred on the C. cupreus lineage. Our results support the hypothesis that karyological evolution in titi monkeys has resulted in reduction in diploid number and that species with higher diploid numbers (with less derived, more ancestral karyotypes)are localized in the centre of the geographic range of the genera, while more derived species appear to occupy the periphery. PMID- 15791415 TI - Populations of a Connecticut River midge structured by geological history and downstream gene flow. AB - Connecticut River midges in the genus Axarus are constrained to semi-isolated populations associated with submerged clay exposures, and gene flow between populations is restricted. I sampled larval midges from fifteen spatially separated sites in the Connecticut River and determined karyotypes for 3111 larvae in two closely related species. In one of these species, an undescribed species referred to as Axarus species varvestris, 5 chromosomal rearrangements were present as polymorphisms. Four of these are paracentric inversions, and the fifth is a terminal insertion or deletion. I used chromosomal rearrangement frequencies to calculate Nei's genetic distance measures for all pairwise population comparisons and I constructed a neighbor-joining phenetic tree for the populations using these distances. Genetic distance between populations of A. sp. varvestris increases with geographic separation, and tree structure indicates that restricted gene flow only occurs in a downstream direction. The effect of unidirectional (downstream) gene flow on population genetic structure in riverine midges is described here for the first time. A genetic break is apparent between populations associated with 12 000-year-old sediments originating in different ancient lakes. The geological break between lakes also coincides with a break in the species composition of the Axarus populations. PMID- 15791419 TI - [Sinusitis. An illness to be taken seriously]. AB - Sinusitis (which should more accurately be called rhinosinusitis because of regular involvement of the nasal cavities) is one of the most widespread illnesses in Europe and the USA, with considerable socioeconomic burden and an increasing prevalence. From a clinical point of view, a distinction is made between acute, acute recurrent and chronic sinusitis. For the patients affected, sinusitis is associated with irritating to agonizing symptoms, which can have a seriously adverse effect on the quality of life. If complications arise, major risks may be involved affecting the orbits and neurocranium. Rational, evidence based diagnosis and therapy is therefore absolutely essential. Guidelines may be helpful, particularly since data available on medication-based treatment, especially of the chronic form, are extremely sparse. PMID- 15791420 TI - [Cortical amaurosis and status epilepticus with acute porphyria]. AB - The most common neurologic manifestations of acute intermittent porphyria (AIP) are autonomic visceral neuropathy, peripheral motor neuropathy, and CNS dysfunctions including seizures and neuropsychiatric disturbances. In rare instances, however, AIP patients have presented with acute cortical blindness. We present a 20-year-old woman who suffered her first attack of AIP. Following 1 week of abdominal pain, she was transferred from a surgical department because of sudden visual loss and deterioration of consciousness. On admission, she developed several generalized seizures. Magnetic resonance imaging showed bilateral DWI lesions occipitally and in the left anterior circulation. Cerebrospinal fluid, MR angiography, and duplex ultrasound were normal. On the following day, sedation and intubation became necessary because of a generalized status epilepticus. Analysis of porphyrinogens in blood, urine and stool showed significantly elevated values. Intravenous therapy with ham-arginate was initiated and antiepileptic therapy was changed to gagabentine. Under this therapeutical regime she remained stable and extubation was possible 48 h later. PMID- 15791418 TI - Sex differences in muscle fatigability and activation patterns of the human quadriceps femoris. AB - The purposes of this study were to determine if the fatigability of the quadriceps femoris varies by biological sex under conditions of normal muscle blood flow and ischemia, and if differences in neuromuscular activation patterns exist. Young men and women (n = 11/group; age 20-39 years) performed a sustained knee extension contraction at 25% of maximal force under conditions of occluded (OCC) and normal muscle blood flow (NON-OCC). Electromyographic (EMG) activity was recorded from the vastus lateralis (VL), rectus femoris (RF), vastus medialis (VM) and biceps femoris (BF) muscles, and analyzed for fatigue-induced changes in the amplitude and burst rate and duration (transient changes in motor unit recruitment) of the signal. Additionally, force fluctuations during the sustained contraction were quantified. Women had a longer time to task failure during the NON-OCC task [214.9 +/- 20.5 vs. 169.1 +/- 20.5 (SE) s] (P = 0.02), but not during the OCC task (179.6 + 19.6 vs. 165.2 +/- 19.6 s). EMG data demonstrated sex differences in the neuromuscular activation pattern of the RF muscle and the collectively averaged QF muscles. During the NON-OCC and OCC tasks women achieved a higher relative activation of the RF at task failure than men (NON-OCC: 40.68 +/- 4.57 vs. 24.49 +/- 4.19%; OCC: 36.80 +/- 5.45 vs. 24.41 +/- 2.12%) (P = 0.02 and 0.05, respectively). Also, during both tasks, they demonstrated a greater relative activation at task failure than men when an average of the VL, VM and RF was considered. Additionally, women exhibited a greater coefficient of variation in force fluctuations during the last-third of the fatiguing NON-OCC task (6.21 +/- 0.567 vs. 4.56 +/- 0.56%) (P = 0.001). No sex differences in EMG burst rate or duration were observed, although there was a trend towards greater EMG burst rate of the RF in association with muscle fatigue in the women (P = 0.09). Interestingly, the only neuromuscular activation variable that displayed a significant relationship with the time to task failure was the average relative EMG of the QF at task failure, and this relationship was observed under both experimental blood flow conditions (NON-OCC: r = 0.47, P = 0.03; OCC: r = 0.44, P = 0.04). These results indicate that sex differences in muscle blood flow and/or muscle metabolism are in part responsible for the female advantage in fatigue resistance. Additionally, these findings suggest that men synergistically recruit the RF compartment to a lesser extent than women in association with muscle fatigue, and that women achieve an overall greater relative activation of the QF at task failure than men. However, the implications of these sex differences in neuromuscular activation patterns during fatiguing muscular contractions on the ability to withstand muscle fatigue (prolonged time to task failure) does not appear to be causally related. PMID- 15791422 TI - Developmental events leading to peltate leaf structure in Tropaeolum majus (Tropaeolaceae) are associated with expression domain changes of a YABBY gene. AB - Peltate leaf architecture has evolved from conventional bifacial leaves many times in flowering plant evolution. Characteristics of peltate leaves, such as the differentiation of a cross zone and of a radially symmetric, margin-less petiole, have also been observed in mutants of genes responsible for adaxial abaxial polarity establishment. This suggests that altered regulation of such genes provided a mechanism for the evolution of peltate leaf structure. Here, we show that evolution of leaf peltation in Tropaeolum majus, a species distantly related to Arabidopsis thaliana, was associated with altered expression of Tropaeolum majus FILAMENTOUS FLOWER (TmFIL), a gene conferring abaxial identity. In situ hybridization indicates that adaxial and abaxial domains are established in early leaf primordia as in species with bifacial leaves. Upon initiation of the cross zone by fusion of the blade margins, localized expansion of TmFIL to the upper leaf side could be seen, indicating a local loss of adaxial leaf identity. The observed changes in expression are consistent with a role of TmFIL in radialization of the petiole and circularization of the leaf blade margin by the cross zone. In addition, expression was observed in segment primordia and during expansion of the bifacial blade, suggesting additional roles for TmFIL in leaf development. PMID- 15791421 TI - Squid is required for efficient posterior localization of oskar mRNA during Drosophila oogenesis. AB - The nuclear-cytoplasmic shuttling heterogeneous nuclear RNA-binding protein (hnRNP) Squid (Sqd) is required during Drosophila melanogaster oogenesis, where it plays a critical role in the regulation of the TGFalpha-like molecule Gurken (Grk). Three Sqd isoforms have been described, SqdA, S and B, and two of these, SqdA and SqdS, differentially function in grk mRNA nuclear export, cytoplasmic transport and translational control during oogenesis. Here, we report that Sqd is also required for the regulation of oskar (osk) mRNA, functioning in the cytoplasmic localization of the osk transcript. In oocytes from sqd females, osk mRNA is not efficiently localized to the posterior pole, but rather accumulates at the anterior cortex. Furthermore, anterior patterning defects observed in embryos from sqd females expressing only the SqdS protein isoform suggest that Sqd may also play a role in the translational regulation of the mislocalized osk mRNA. These findings provide additional support for models of mRNA regulation in which cytoplasmic events, such as localization and translational regulation, are coupled. These results also place Sqd among an emerging class of proteins, including such other members as Bruno (Bru) and Hrb27C/Hrp48, which function in multiple aspects of both grk and osk mRNA regulation during Drosophila oogenesis. PMID- 15791423 TI - Skin tension and cleavage lines (Langer's lines) causing distortion of ante- and postmortem wound morphology. AB - The assessment of individual wounds at autopsy may be complicated by the superimposition of a number of injuries or damage to tissues that occurred after death, either of which has the potential to distort the morphology of the initial injury. Additional factors that may change the shape of wounds are (1) the relationship of the wound to the so-called skin cleavage lines (Langer's lines) and (2) tension placed on the skin. Three autopsy cases are reported to demonstrate once more how wound morphology may be altered by such factors. In case 1, rectangular stab wounds to the base of the neck in a 53-year-old man, which suggested that a square or rectangular tool may have caused the injuries, were altered to more typical knife stab wounds once skin tension had been released at autopsy. The uppermost wounds, however, continued to gape due to the effects of skin cleavage lines. In case 2, slit-like wounds resembling stab wounds in the neck of a 54-year-old woman found in a river were shown to be circular once skin tension had been released. In case 3, the effects of either cleavage lines or skin tension could be demonstrated on excised wounds from a 43 year-old man whose body had also been found in a river; tensile forces easily changed circular into slit-like wounds. Tension and/or skin cleavage lines may transform round skin defects into slit-like wounds resembling knife stab wounds, round out genuine stab wounds and artefactually lengthen stab wounds. These factors must be taken into consideration carefully when wounds are assessed at the death scene prior to autopsy. PMID- 15791424 TI - Dynamics of leaf area and nitrogen in the canopy of an annual herb, Xanthium canadense. AB - We studied leaf area and nitrogen dynamics in the canopy of stands of an annual herb Xanthium canadense, grown at a high (HN)- and a low-nitorgen (LN) availability. Standing leaf area increased continuously through the vegetative growth period in the LN stand, or leveled off in the later stage in the HN stand. When scaled against standing leaf area, both production and loss rates of leaf area increased but with different patterns: the production rate was retarded, while the loss rate was accelerated, implying an upper limit of standing leaf area of the canopy. The rate of leaf-area production was higher in the HN than in the LN stand, which was caused by the higher rate of leaf production per standing leaf area as well as the greater standing leaf area in the HN stand. Although the rate of leaf-area loss was higher in the HN than in the LN stand, it was not significantly different between the two stands when compared at a common standing leaf area, suggesting involvement of light climate in determination of the leaf loss rate. On the other hand, the rate of leaf-area loss was positively correlated with nitrogen demand for leaf area development across the two stands, suggesting that leaf loss was caused by retranslocation of nitrogen for construction of new leaves. A simple simulation model of leaf and nitrogen dynamics in the canopy showed that, at steady state, where the rate of leaf-area loss becomes equal to the production rate, the standing leaf area was still greater in the HN than in the LN stand. Similarly, when the uptake and loss of nitrogen are equilibrated, the standing nitrogen was greater in the HN than in the LN stand. These results suggest that leaf-area production is strongly controlled by nitrogen availability, while both nitrogen and light climate determine leaf-loss rates in the canopy. PMID- 15791425 TI - Induced plant responses to multiple damagers: differential effects on an herbivore and its parasitoid. AB - Herbivore-induced plants responses can affect the preference and performance of herbivores and their natural enemies. These responses may vary depending on the identity and number of herbivore species feeding on the plant so that when herbivores from different guilds feed on plants, the interactions between plants, herbivores, and natural enemies may be disrupted. Tomato plants were damaged either by the caterpillar Spodoptera exigua, or the aphid Macrosiphum euphorbiae, or damaged by both herbivores, or undamaged controls. We measured the preference and performance of S. exigua and its parasitoid Cotesia marginiventris, and activity of proteinase inhibitors (PI) as an indicator of induced resistance. Compared to undamaged plants, caterpillar damage reduced the number of eggs laid by S. exigua adults, reduced growth, consumption, and survival of larval S. exigua and C. marginiventris, and increased activity of PIs 43%; but did not increase attraction of C. marginiventris. While pupal mass of S. exigua was not affected, the pupal mass of C. marginiventris decreased on caterpillar-damaged plants compared to controls. In contrast, plants damaged by aphids were preferred for oviposition by S. exigua, and had increased larval consumption and survival, compared to controls. Aphid feeding did not affect the preference or performance of C. marginiventris, or PI activity, compared to controls. While oviposition was deterred on caterpillar-damaged plants, plants damaged by both herbivores received the same amount of oviposition as controls. The attraction of C. marginiventris to plants damaged by caterpillars and aphids was increased compared to controls. However, plants damaged by both herbivores had similar PI activity, larval growth and survival of S. exigua and C. marginiventris, as plants singly damaged by caterpillars. Overall, the preference component for both the herbivore and parasitoid was more strongly affected by damage due to multiple herbivores than the performance component. PMID- 15791426 TI - Differential effects of lichens, mosses and grasses on respiration and nitrogen mineralization in soils of the New Jersey Pinelands. AB - In the New Jersey Pinelands, severely disturbed areas often do not undergo a rapid succession to forest; rather, a patchy cover of lichens, mosses and grasses persists for decades. We hypothesized that these plant covers affect soil microbial processes in different ways, and that these effects may alter the successional dynamics of the patches. We predicted that the moss and grass covers stimulate soil microbial activity, whereas lichens inhibit it, which may in turn inhibit succession. We collected soil cores from beneath each type of cover plus bare soil within two types of highly disturbed areas--sites subjected to hot wildfires, and areas mined for sand. Organic matter (OM) content, soil respiration and potential N mineralization were measured in the cores. Soils under mosses were similar to those under grasses; they accumulated more OM and produced more mineral N, predominantly in the form of ammonium, than either the bare soils or the soils beneath lichens. Mineralization under lichens, like that of the bare soils but unlike the soils beneath mosses or grasses, was dominated by net nitrification. These patterns were reproduced in experimentally transplanted moss and lichen mats. Mosses appear to create high-nutrient microsites via high rates of OM accumulation and production of ammonium, whereas lichens maintain low-nutrient patches similar to bare soil via low OM accumulation rates and production of mineral N predominantly in the mobile nitrate form. These differences in soil properties may explain the lack of vascular plant invasion in lichen mats, in contrast to the moss-dominated areas. PMID- 15791428 TI - Relative performance of European grapevine moth (Lobesia botrana) on grapes and other hosts. AB - The European grapevine moth, Lobesia botrana is a major grapevine pest, but despite the abundance of vineyards it is a generalist and uses either grapes or alternative species. Given the abundance and predictability of grape, L. botrana could be expected to have evolved towards monophagy. In order to understand why this species remains polyphagous, we hypothesized that larvae reared on rare wild host plants should have higher fitness than those reared on the more abundant grape host. For this, we compared larval performance and several life history traits on three alternative host plants (Daphne gnidium, Olea europaea, Tanacetum vulgare) and three Vitaceae (Vitis vinifera), two cultivars and one wild species (Ampelopsis brevipedunculata), and two control groups raised on either a low or a high nutritive value medium. Alternative hosts are more suitable than Vitaceae for the reproductive performance of L. botrana: larval mortality and development time was reduced, while pupal weight, growth rate, female longevity, female fecundity, duration of laying and mating success were increased. High quality food ingested by larvae promotes higher adult body weight and enhances female reproductive output. This suggests that alternative hosts provide greater nutritional value for L. botrana than Vitaceae. The use of alternative host plants could thus be maintained in the host range because they offer L. botrana a better fitness than on the Vitaceae. This could typically represent an advantage for moths behaving in plant diversity grape landscapes. PMID- 15791427 TI - Genotype x environment interactions, stoichiometric food quality effects, and clonal coexistence in Daphnia pulex. AB - The role of stoichiometric food quality in influencing genotype coexistence and competitive interactions between clones of the freshwater microcrustacean, Daphnia pulex, was examined in controlled laboratory microcosm experiments. Two genetically distinct clones of D. pulex, which show variation in their ribosomal rDNA structure, as well as differences in a number of previously characterized growth-rate-related features (i.e., life-history features), were allowed to compete in two different arenas: (1) batch cultures differing in algal food quality (i.e., high vs. low carbon:phosphorus (C:P ratio) in the green alga, Scenedesmus acutus); (2) continuous flow microcosms receiving different light levels (i.e., photosynthetically active radiation) that affected algal C:P ratios. In experiment 1, a clear genotype x environment interaction was determined with clone 1 out-competing clone 2 under high nutrient (i.e., low food C:P) conditions, while the exact opposite pattern was observed under low nutrient (i.e., high C:P) conditions. In experiment 2, clone 1 dominated over clone 2 under high light (higher C:P) conditions, but clonal coexistence was observed under low light (low C:P) conditions. These results indicate that food (nutrient) quality effects (hitherto an often overlooked factor) may play a role in microevolutionary (genotypic) responses to changing stoichiometric conditions in natural populations. PMID- 15791429 TI - Predation regulation of sedimentary faunal structure: potential effects of a fishery-induced switch in predators in a Newfoundland sub-Arctic fjord. AB - The collapse of the cod fishery in Newfoundland has coincided with marked increases in abundances of snow crab, pandalid shrimp, and other crustaceans that prey on sedimentary infauna. A 3-year sampling program in Bonne Bay, Newfoundland indicates differences in composition and number of these predators in the two main arms of the fjord that coincide with strong differences in benthic community structure. To test whether predation pressure contributes to the observed patterns in sedimentary fauna, exclusion field experiments with full and partial cages were deployed in both arms at 30-m depth and sampled along with ambient sediments at 0-, 4-, and 8-week periods. Predation significantly influenced species composition, abundance and, in some cases, diversity. The most striking changes included increases in the polychaetes Pholoe tecta and Ophelina cylindricaudata in exclusions relative to controls, and concurrent declines in the polychaete Paradoneis lyra and the cumacean Lamphros fuscata. In laboratory experiments, fresh non-disturbed sediment cores from each experimental area were either protected or exposed to snow crab, the most abundant predator in the bay. A snow crab inclusion experiment was also carried out in the field, using cages similar to those used for exclusions. Despite differences in sedimentary faunas in the two arms, both types of experiments detected a predator effect that was very similar to that documented in exclusion experiments. Thus, despite differences in the scales associated with each type of manipulation, our results suggest that crab predation is a significant structuring force in Newfoundland sedimentary communities. Given the historical changes that have occurred in predator composition as a result of cod over-fishing, we hypothesize that broad scale community changes may be taking place in North Atlantic benthic ecosystems. PMID- 15791430 TI - Niche differences in phenology and rooting depth promote coexistence with a dominant C4 bunchgrass. AB - Spatial and temporal niche differentiation are potential mechanisms of plant species' coexistence in many communities, including many grasslands. In a 6-year field experiment, a dominant prairie bunchgrass, Schizachyrium scoparium, excluded species sharing similar phenology and rooting depth, but coexisted with species differing in phenology and rooting depth. We used a series of experimental plots that differed in S. scoparium abundance to measure the effect of S. scoparium on abundances of other species and on soil nitrate concentrations across time and depth. Because we found that S. scoparium most strongly reduced soil nitrate levels at mid-season and at shallow depths, we hypothesized that at higher S. scoparium abundance, species with mid-season phenology and shallow roots would be excluded and that deep-rooted and early season species would be more likely to persist. As predicted, the proportional number and cover of species exploiting different niches than S. scoparium (early season and deep rooted species) increased across the S. scoparium abundance gradient. This work provides novel empirical evidence for the factors that contribute to species coexistence in the field. Our study suggests that spatial and temporal niche differentiation promote species coexistence in these grasslands. PMID- 15791431 TI - Game theory provides no explanation for seed size variation in grasslands. AB - Game theoretical models have been suggested to explain the maintenance of a remarkable variation in seed size across species in most types of vegetation. According to these models, which are based on the existence of a trade-off between seed size and seed number; smaller-seeded species can invade any species mixture due to their numerical advantage, and larger-seeded species can invade any species mixture due to their competitive superiority over smaller-seeded species during recruitment. However, till now, there is very limited evidence for seed size effects on recruitment interactions among different species. An experiment was designed using 16 species in Swedish grasslands, varying 384-fold in seed size. Species were sown pairwise and alone in disturbed versus undisturbed small plots in grazed versus ungrazed grassland, and the resulting recruitment was recorded. Seedling densities in the experimental plots were within the natural range. Both disturbance and grazing had a positive effect on recruitment. Seed size did not affect recruitment except from a tendency that increasing seed size made recruitment less dependent on disturbance. Recruitment of smaller-seeded species was not affected by the presence of larger-seeded species. Larger-seeded species did not generally win in direct contest. These results suggest that game theoretical models do not explain maintenance of seed size variation across species in these grasslands. Alternative explanations for seed size variation are that either small-scale heterogeneity provide conditions favouring a range of different seed sizes or other attributes than seed size effectively determine recruitment. PMID- 15791432 TI - PET imaging of brain with the beta-amyloid probe, [11C]6-OH-BTA-1, in a transgenic mouse model of Alzheimer's disease. AB - PURPOSE: The purpose of this study was to evaluate the capacity of [11C]6-OH-BTA 1 and positron emission tomography (PET) to quantify beta-amyloid (Abeta) plaques in the Tg2576 mouse model of Alzheimer's disease (AD). METHODS: PET imaging was performed with the NIH ATLAS small animal scanner in six elderly transgenic mice (Tg2576; age 22.0+/-1.8 months; 23.6+/-2.6 g) overexpressing a mutated form of human beta-amyloid precursor protein (APP) known to result in the production of Abeta plaques, and in six elderly wild-type litter mates (age 21.8+/-1.6 months; 29.5+/-4.7 g). Dynamic PET scans were performed for 30 min in each mouse under 1% isoflurane inhalation anesthesia after a bolus injection of 13-46 MBq of [11C]6 OH-BTA-1. PET data were reconstructed with 3D OSEM. On the coronal PET image, irregular regions of interest (ROIs) were placed on frontal cortex (FR), parietal cortex (PA), striatum (ST), thalamus (TH), pons (PO), and cerebellum (CE), guided by a mouse stereotaxic atlas. Time-activity curves (TACs) (expressed as percent injected dose per gram normalized to body weight: % ID-kg/g) were obtained for FR, PA, ST, TH, PO, and CE. ROI-to-CE radioactivity ratios were also calculated. Following PET scans, sections of mouse brain prepared from anesthetized and fixative-perfused mice were stained with thioflavin-S. RESULTS: TACs for [11C]6 OH-BTA-1 in all ROIs peaked early (at 30-55 s), with radioactivity washing out quickly thereafter in both transgenic and wild-type mice. Peak uptake in all regions was significantly lower in transgenic mice than in wild-type mice. During the later part of the washout phase (12-30 min), the mean FR/CE and PA/CE ratios were higher in transgenic than in wild-type mice (1.06+/-0.04 vs 0.98+/-0.07, p=0.04; 1.06+/-0.09 vs 0.93+/-0.08 p=0.02) while ST/CE, TH/CE, and PO/CE ratios were not. Ex vivo staining revealed widespread Abeta plaques in cortex, but not in cerebellum of transgenic mice or in any brain regions of wild-type mice. CONCLUSION: Marked reductions in brain uptake of this radioligand in transgenic mice may be due to reduced cerebral blood flow relative to that in wild-type mice. Specific [11C]6-OH-BTA-1 binding to Abeta plaques, if any, is probably very low, as reflected in the small FR/CE and PA/CE ratio differences. FR/CE and PA/CE ratios are considerably higher in AD patients while Abeta plaque densities in 22 month-old transgenic mice may be expected to show essentially the same density as is observed in the AD brain. This implies that the absence of tracer retention in 22-month-old transgenic mice may be due to the smaller number of Abeta plaque binding sites and/or to lower affinity of the binding sites for [11C]6-OH-BTA-1 as compared with AD patients. [11C]6-OH-BTA-1 shows excellent brain uptake in mice. PMID- 15791433 TI - Clinical axillary recurrence after sentinel node biopsy in breast cancer: a follow-up study of 220 patients. AB - PURPOSE: The aim of this study was to evaluate the frequency of false-negative (FN) sentinel node procedures in patients with breast cancer and the subsequent clinical outcome in such patients. METHODS: A total of 325 breast cancer patients underwent sentinel lymph node biopsy at our institution between June 1998 and May 2004. A 2-day protocol was used to localise the sentinel node with the injection of 99mTc-nanocolloid. There were two phases in the study: the learning phase (105 patients) and the application phase (220 patients). In the learning phase, a complete lymphadenectomy was always performed. In the application phase, sentinel nodes were studied intraoperatively and lymphadenectomy was performed when considered warranted by the pathological intraoperative results. RESULTS: The median follow-up duration in the 220 patients studied during the application phase was 21.2 months (range 4-45 months). In this phase a total of 427 sentinel nodes were obtained (range 1-5 per patient, median 1.99), with 66 positive sentinel nodes in 56 patients (26%). The lymphadenectomies performed were also positive in 25% of cases (14 patients). We observed a total of two false-negative sentinel lymph node results (3.45%). One of them was found during the surgical excision of non-sentinel nodes, and the other presented as an axillary recurrence 17 months postoperatively (1.72% clinical false-negative rate). The latter patient died 1 year after the first recurrence. CONCLUSION: After a median follow up of 21.2 months we observed only one clinical recurrence among 220 patients. Our results indicate that adequate local control is achieved by application of the sentinel node protocol. PMID- 15791434 TI - The uptake of 3'-deoxy-3'-[18F]fluorothymidine into L5178Y tumours in vivo is dependent on thymidine kinase 1 protein levels. AB - PURPOSE: The aim of this study was to investigate the role of thymidine kinase 1 (TK1) protein in 3'-deoxy-3'-[18F]fluorothymidine ([18F]FLT) positron emission tomography (PET) studies. METHODS: We investigated the in vivo kinetics of [18F]FLT in TK1+/- and TK1-/- L5178Y mouse lymphoma tumours that express different levels of TK1 protein. RESULTS: [18F]FLT-derived radioactivity, measured by a dedicated small animal PET scanner, increased within the tumours over 60 min. The area under the normalised tumour time-activity curve were significantly higher for the TK1+/- compared with the -/- variant (0.89+/-0.02 vs 0.79+/-0.03 MBq ml(-1) min, P=0.043; n=5 for each tumour type). Ex vivo gamma counting of tissues excised at 60 min p.i. (n=8) also revealed significantly higher tumour [18F]FLT uptake for the TK1+/- variant (6.2+/-0.6 vs 4.6+/-0.4%ID g(-1), P=0.018). The observed differences between the cell lines with respect to [18F]FLT uptake were in keeping with a 48% higher TK1 protein in the TK1+/- tumours versus the -/- variant (P=0.043). On average, there were no differences in ATP levels between the two tumour variants (P=1.00). A positive correlation between [18F]FLT accumulation and TK1 protein levels (r=0.68, P=0.046) was seen. Normalisation of the data for ATP content further improved the correlation (r=0.86, P=0.003). CONCLUSION: This study shows that in vivo [18F]FLT kinetics depend on TK1 protein expression. ATP may be important in realising this effect. Thus, [18F]FLT-PET has the potential to yield specific information on tumour proliferation in diagnostic imaging and therapy monitoring. PMID- 15791435 TI - Pharmacokinetics and biodistribution of 177Lu-labeled multivalent single-chain Fv construct of the pancarcinoma monoclonal antibody CC49. AB - PURPOSE: Lutetium-177 (177Lu) is a radionuclide of interest for radioimmunoimaging (RII) and radioimmunotherapy (RIT) on account of its short half-life (161 h) and the ability to emit both beta and gamma radiation. Single chain Fv (scFv) constructs have shown advancement in cancer diagnosis and therapy due to the pharmacokinetics advantage and seem to be intriguing tools in oncology. The objective of this study was to evaluate the pharmacokinetics and biodistribution characteristics of the 177Lu-labeled tetravalent scFv of CC49 MAb and intact CC49 IgG in vivo. METHODS: Conjugation and labeling conditions of multivalent scFv with 177Lu were optimized without affecting integrity and immunoreactivity. For this purpose, multivalent scFv constructs {dimer, sc(Fv)2; tetramer, [sc(Fv)2]2} of the MAb CC49 were expressed as secretory proteins in Pichia pastoris. The purified scFv constructs and IgG form of CC49 were conjugated with a bifunctional chelating agent, ITCB-DTPA, and labeled with 177Lu. The comparative biodistribution, blood clearance, and tumor-targeting characteristics of 177Lu-labeled tetravalent [sc(Fv)2]2 construct of CC49 MAb and intact CC49 IgG were investigated in the athymic mice bearing LS-174T xenografts. RESULTS: Approximately, 90% of 177Lu incorporation was achieved using ITCB-DTPA chelator, and the labeled immunoconjugates maintained integrity and immunoreactivity. Blood clearance studies demonstrated an alpha half-life (t1/2alpha) of 177Lu-labeled [sc(Fv)2]2 and IgG of CC49 at 4.40 and 9.50 min and a beta half-life (t1/2beta) at 375 and 2,193 min, respectively. At 8 h post administration, the percent of the injected dose accumulated/gram (%ID/g) of the LS-174T tumor was 6.4+/-1.3 and 8.9+/-0.6 for 177Lu-labeled [sc(Fv)2]2 and IgG of CC49, respectively, in the absence of L-lysine. The corresponding values were 8.0+/-0.6 and 8.4+/-1.2 in the presence of L-lysine. Renal accumulation of [sc(Fv)2]2 was significantly (p<0.005) reduced in the presence of L-lysine. CONCLUSION: The results of this study demonstrate that the ITCB-DTPA conjugation and 177Lu-labeling of scFvs are feasible without influencing the antibody characteristics. 177Lu-labeled [sc(Fv)2]2 showed faster clearance and equivalent tumor uptake at 8 h compared with its IgG form, with a markedly reduced renal uptake in the presence of L-lysine. Therefore, 177Lu-labeled [sc(Fv)2]2 may be a potential radiopharmaceutical for the treatment of cancer. PMID- 15791436 TI - Cell death triggered by alpha-emitting 213Bi-immunoconjugates in HSC45-M2 gastric cancer cells is different from apoptotic cell death. AB - PURPOSE: Radioimmunotherapy with alpha-particle-emitting nuclides, such as 213Bi, is a promising concept for the elimination of small tumour nodules or single disseminated tumour cells. The aim of this study was to investigate cellular damage and the mode of cell death triggered by 213Bi-immunoconjugates. METHODS: Human gastric cancer cells (HSC45-M2) expressing d9-E-cadherin were incubated with different levels of activity of 213Bi-d9MAb targeting d9-E-cadherin and 213Bi-d8MAb, which does not bind to d9-E-cadherin. Micronucleated (M) cells, abnormal (A) cells and apoptotic (A) [(MAA)] cells were scored microscopically in the MAA assay following fluorescent staining of nuclei and cytoplasm. Chromosomal aberrations were analysed microscopically following Giemsa staining. The effect of z-VAD-fmk, known to inhibit apoptosis, on the prevention of cell death was investigated following treatment of HSC45-M2 cells with sorbitol as well as 213Bi d9MAb. Activation of caspase 3 after incubation of HSC45-M2 cells with both sorbitol and 213Bi-d9MAb was analysed via Western blotting. RESULTS: Following incubation of HSC45-M2 human gastric cancer cells expressing d9-E-cadherin with 213Bi-d9MAb the number of cells killed increased proportional to the applied activity concentration. Microscopically visible effects of alpha-irradiation of HSC45-M2 cells were formation of micronuclei and severe chromosomal aberrations. Preferential induction of these lesions with specific 213Bi-d9MAb compared with unspecific 213Bi-d8MAb (not targeting d9-E-cadherin) was not observed if the number of floating, i.e. unbound 213Bi-immunoconjugates per cell exceeded 2 x 10(4), most likely due to intense crossfire. In contrast to sorbitol-induced cell death, cell death triggered by 213Bi-immunoconjugates was independent of caspase 3 activation and could not be inhibited by z-VAD-fmk, known to suppress the apoptotic pathway. CONCLUSION: 213Bi-immunoconjugates seem to induce a mode of cell death different from apoptosis in HSC45-M2 cells. PMID- 15791437 TI - Factors of importance for scintigraphic non-visualisation of sentinel nodes in breast cancer. AB - PURPOSE: The aim of this study was to analyse different factors of possible significance for non-visualisation of sentinel nodes (SNs) by preoperative lymphoscintigraphy, in order to enable improvement of the success rate of SN visualisation through modification or alteration of some of the factors. METHODS: Between March 1998 and January 2003 we analysed a series of 442 women with unilateral stage T1 and clinical N0 breast cancer. Lymphoscintigraphy was performed after periareolar or peritumoural injection of 99mTc-albumin nanocolloid, with image acquisition after 2-6 h or 18-24 h. Until January 2001, all patients received around 20 MBq tracer, irrespective of time to operation. From January 2001, patients injected on the day before surgery received at least 100 MBq while patients injected on the day of surgery received around 50 MBq. RESULTS: An SN was visualised in 87% of the patients, and at surgery the SN was detected with the hand-held gamma probe in 42% of the remaining patients. By multiple logistic regression analysis, statistically significant independent variables that increased the risk for non-visualisation were increasing age (p=0.0007), increasing body weight (p=0.0189) and peritumoural injection (p<0.0001). Significant interaction was found for imaging time and injected activity (p=0.0017). CONCLUSION: This study conclusively shows that the risk of unsuccessful SN imaging increases with age and body weight. Our findings suggest that the scintigraphic success rate may be improved by periareolar (rather than peritumoural) injection. Early and late imaging procedures are equally efficient, but if a late imaging procedure is used, activity (adjusted for physical decay) in the patient on day 2 should be more than 10 MBq. PMID- 15791438 TI - Effects of ROI definition and reconstruction method on quantitative outcome and applicability in a response monitoring trial. AB - PURPOSE: Quantitative measurement of tracer uptake in a tumour can be influenced by a number of factors, including the method of defining regions of interest (ROIs) and the reconstruction parameters used. The main purpose of this study was to determine the effects of different ROI methods on quantitative outcome, using two reconstruction methods and the standard uptake value (SUV) as a simple quantitative measure of FDG uptake. METHODS: Four commonly used methods of ROI definition (manual placement, fixed dimensions, threshold based and maximum pixel value) were used to calculate SUV (SUV([MAN]), SUV15 mm, SUV50, SUV75 and SUVmax, respectively) and to generate "metabolic" tumour volumes. Test-retest reproducibility of SUVs and of "metabolic" tumour volumes and the applicability of ROI methods during chemotherapy were assessed. In addition, SUVs calculated on ordered subsets expectation maximisation (OSEM) and filtered back-projection (FBP) images were compared. RESULTS: ROI definition had a direct effect on quantitative outcome. On average, SUV[MAN), SUV15 mm, SUV50 and SUV75, were respectively 48%, 27%, 34% and 15% lower than SUVmax when calculated on OSEM images. No statistically significant differences were found between SUVs calculated on OSEM and FBP reconstructed images. Highest reproducibility was found for SUV15 mm and SUV[MAN] (ICC 0.95 and 0.94, respectively) and for "metabolic" volumes measured with the manual and 50% threshold ROIs (ICC 0.99 for both). Manual, 75% threshold and maximum pixel ROIs could be used throughout therapy, regardless of changes in tumour uptake or geometry. SUVs showed the same trend in relative change in FDG uptake after chemotherapy, irrespective of the ROI method used. CONCLUSION: The method of ROI definition has a direct influence on quantitative outcome. In terms of simplicity, user-independence, reproducibility and general applicability the threshold-based and fixed dimension methods are the best ROI methods. Threshold methods are in addition relatively independent of changes in size and geometry, however, and may therefore be more suitable for response monitoring purposes. PMID- 15791440 TI - Quantitation of dopamine transporter blockade by methylphenidate: first in vivo investigation using [123I]FP-CIT and a dedicated small animal SPECT. AB - PURPOSE: The aim of this study was to investigate the feasibility of assessing dopamine transporter binding after treatment with methylphenidate in the rat using a recently developed high-resolution small animal single-photon emission computed tomograph (TierSPECT) and [123I]FP-CIT. METHODS: [123I]FP-CIT was administered intravenously 1 h after intraperitoneal injection of methylphenidate (10 mg/kg) or vehicle. Animals underwent scanning 2 h after radioligand administration. The striatum was identified by superimposition of [123I]FP-CIT scans with bone metabolism and perfusion scans obtained with 99mTc-DPD and 99mTc tetrofosmin, respectively. As these tracers do not pass the blood-brain barrier, their distribution permits the identification of extracerebral anatomical landmarks such as the orbitae and the harderian glands. The cerebellum was identified by superimposing [123I]FP-CIT scans with images of brain perfusion obtained with 99mTc-HMPAO. RESULTS: Methylphenidate-treated animals and vehicle treated animals yielded striatal equilibrium ratios (V''3) of 0.24+/-0.26 (mean +/- SD) and 1.09+/-0.42, respectively (t test, two-tailed, p<0.0001). Cortical V''3 values amounted to 0.05+/-0.28 (methylphenidate) and 0.3+/-0.39 (saline, p=0.176). This first in vivo study of rat dopamine transporter binding after pre treatment with methylphenidate showed a mean reduction of 78% in striatal [123I]FP-CIT accumulation. CONCLUSION: The results can be interpreted in terms of a pharmacological blockade in the rat striatum and show that in vivo quantitation of dopamine transporter binding is feasible with [123I]FP-CIT and the TierSPECT. This may be of future relevance for in vivo investigations on rat models of attention deficit/hyperactivity disorder. Furthermore, our findings suggest that investigations in other animal models, e.g. of Parkinson's and Huntington's disease, may be feasible using SPECT radioligands and small animal imaging systems. PMID- 15791439 TI - Differences in [99mTc]TRODAT-1 SPECT binding to dopamine transporters in patients with multiple system atrophy and Parkinson's disease. AB - PURPOSE: Multiple system atrophy (MSA), a disorder causing autonomic dysfunction, parkinsonism, and cerebellar dysfunction, is difficult to differentiate from other movement disorders, particularly early in the course of disease. This study evaluated whether [99mTc]TRODAT-1 binding to the dopamine transporter differentiates MSA from other movement disorders. METHODS: Single-photon emission computed tomographic brain scans were acquired in 25 MSA patients, 48 age-matched controls, and 130 PD patients, 3 h after the injection of 740 MBq (20 mCi) of [99mTc]TRODAT-1. Regions of interest (ROIs) were placed manually on subregions of both basal ganglia and distribution volume ratios (DVRs) were calculated. Regional DVRs were compared between study groups in MSA patients. Student's t tests were used to compare MSA patients with other study groups. Spearman correlations were used to compare DVRs with NP measures. RESULTS: Based upon various motor scores, MSA and PD patients had comparable motor impairment, and were significantly impaired compared with controls. Mean DVRs in the basal ganglia of MSA patients were significantly less than those of controls, but generally higher (p<0.05) than in PD patients. In particular, the MSA patients had significantly increased DVRs in the posterior putamen (mean 0.49+/-0.30) compared with PD patients (0.74+/-0.25). CONCLUSION: Movement disorder patients could be differentiated from controls, but MSA and PD patients could not be easily differentiated from each other. As a group, MSA patients had significantly higher mean [99mTc]TRODAT-1 binding, particularly in the posterior putamen, compared with PD patients and significantly lower binding compared with controls. This may reflect different pathophysiological processes of the two neurodegenerative diseases. PMID- 15791441 TI - Incidence of stunned, hibernating and scarred myocardium in ischaemic cardiomyopathy. AB - PURPOSE: Different criteria to identify residual viability in chronically dysfunctioning myocardium in patients with coronary artery disease (CAD) can be derived by the combined assessment of myocardial blood flow (MBF) and glucose utilisation (MRG) using positron emission tomography (PET). The aim of this study was to evaluate, in a large number of patients, the prevalence of these different patterns by purely quantitative means. METHODS: One hundred and sixteen consecutive patients with ischaemic cardiomyopathy (LVEF < or =40%) underwent resting 2D echocardiography to assess regional contractile function (16-segment model). PET with 15O-labelled water (H2 15O) and 18F-fluorodeoxyglucose (FDG) was used to quantify MBF and MRG during hyperinsulinaemic euglycaemic clamp. Dysfunctional segments with normal MBF (> or =0.6 ml min(-1) g(-1)) were classified as stunned, and segments with reduced MBF (<0.6 ml min(-1) g(-1)) as hibernating if MRG was > or =0.25 micromol min(-1) g(-1). Segments with reduced MBF and MRG <0.20 micromol min(-1) g(-1) were classified as transmural scars and segments with reduced MBF and MRG between 0.20 and 0.25 micromol min(-1) g(-1) as non-transmural scars. RESULTS: Eight hundred and thirty-four (46%) segments were dysfunctional. Of these, 601 (72%) were chronically stunned, with 368 (61%) having normal MRG (0.47+/-0.20 micromol min(-1) g(-1)) and 233 (39%) reduced MRG (0.16+/-0.05 micromol min(-1) g(-1)). Seventy-four (9%) segments with reduced MBF had preserved MRG (0.40+/-0.18 micromol min(-1) g(-1)) and were classified as hibernating myocardium. In addition, 15% of segments were classified as transmural and 4% as non-transmural scar. The mean MBF was highest in stunned myocardium (0.95+/-0.32 ml min(-1) g(-1)), intermediate in hibernating myocardium and non-transmural scars (0.47+/-0.09 ml min(-1) g(-1) and 0.48+/-0.08 ml min(-1) g(-1), respectively), and lowest in transmural scars (0.40+/-0.14 ml min(-1) g( 1), P<0.01). MRG was comparable in hibernating and stunned myocardium with preserved MRG (0.40+/-0.19 micromol min(-1) g(-1) vs 0.46+/-0.20 micromol min(-1) g(-1), NS), and lowest in stunned myocardium with reduced MRG and transmural scars. CONCLUSION: Chronic stunning is more prevalent than expected. The degree of MRG reduction in stunned myocardium may disclose segments at higher risk of permanent damage. PMID- 15791447 TI - ECG-triggered high-resolution positron emission tomography: a breakthrough in cardiac molecular imaging of mice. PMID- 15791442 TI - Effects of nicorandil on cardiac sympathetic nerve activity after reperfusion therapy in patients with first anterior acute myocardial infarction. AB - PURPOSE: Ischaemic preconditioning (PC) is a cardioprotective phenomenon in which short periods of myocardial ischaemia result in resistance to decreased contractile dysfunction during a subsequent period of sustained ischaemia. Nicorandil, an ATP-sensitive potassium channel opener, can induce PC effects on sympathetic nerves during myocardial ischaemia. However, its effects on cardiac sympathetic nerve activity (CSNA) and left ventricular remodelling have not been determined. In this study, we sought to determine whether nicorandil administration improves CSNA in patients with acute myocardial infarction (AMI). METHODS: We studied 58 patients with first anterior AMI, who were randomly assigned to receive nicorandil (group A) or isosorbide dinitrate (group B) after primary coronary angioplasty. The nicorandil or isosorbide dinitrate was continuously infused for >48 h. The extent score (ES) was determined from 99mTc pyrophosphate scintigraphy, and the total defect score (TDS) was determined from 201Tl scintigraphy 3-5 days after primary angioplasty. The left ventricular end diastolic volume (LVEDV) and left ventricular ejection fraction (LVEF) were determined by left ventriculography 2 weeks later. The delayed heart/mediastinum count (H/M) ratio, delayed TDS and washout rate (WR) were determined from 123I meta-iodobenzylguanidine (MIBG) images 3 weeks later. The left ventriculography results were re-examined 6 months after treatment. RESULTS: Fifty patients originally enrolled in the trial completed the entire protocol. After treatment, no significant differences were observed in ES or left ventricular parameters between the two groups. However, in group A (n=25), the TDSs determined from 201Tl and 123I-MIBG were significantly lower (26+/-6 vs 30+/-5, P<0.01, and 32+/ 8 vs 40+/-6, P<0.0001, respectively), the H/M ratio significantly higher (1.99+/ 0.16 vs 1.77+/-0.30, P<0.005) and the WR significantly lower (36%+/-8% vs 44%+/ 12%, P<0.005) than in group B (n=25). Moreover, 6 months after treatment, LVEDV and LVEF were better in group A than in group B. CONCLUSION: These findings indicate that nicorandil can have beneficial effects on CSNA and left ventricular remodelling in patients with first anterior AMI. PMID- 15791443 TI - Quantification of disease activity in patients undergoing leucocyte scintigraphy for suspected inflammatory bowel disease. AB - PURPOSE: Whole-body gamma camera counting is an alternative to faecal 111In collection for quantifying disease activity in inflammatory bowel disease (IBD) but requires administration of imaging activities of 111In. The aim of this study was to explore a dedicated whole-body counter which requires 20-fold less activity than gamma camera counting. METHODS: Thirty patients with known or suspected IBD received 99mTc-granulocytes (approximately 200 MBq) and 111In granulocytes (approximately 0.5 MBq). The 99mTc-cells were injected 45 min after the 111In-cells and immediately after a baseline 111In whole-body count. The decay-corrected count at 120 h was expressed as a fraction of baseline to give whole-body 111In retention (WBR). One patient was excluded as the injected cells were non-viable. RESULTS: Median 45-min intravascular 111In recovery was 35% in patients compared with 43% in six normal volunteers (p<0.05). WBR in normals ranged from 91% to 93% and in 11 patients with negative 99mTc imaging from 87% to 96%. Only one of 11 patients with negative imaging had WBR <90% while 11/12 patients with WBR <90% had abnormal imaging. Ten of 13 patients with Crohn's disease (CD) had abnormal imaging. The lowest WBR in these ten was 77% and six had values of >90%. There was a significant association between WBR and 99mTc image grade (Rs=0.73, p<0.002) in 16 patients without CD, but not in 13 patients with CD (Rs=0.54, p>0.05). CONCLUSION: Dedicated whole-body counting is able to quantify disease activity in IBD but may give normal values in active CD. PMID- 15791448 TI - 18F-FDG uptake in bilateral adrenal hyperplasia causing Cushing's syndrome. PMID- 15791449 TI - Modeling the acceleration sensitive neurons in the pigeon optokinetic system. AB - Recent physiological findings revealed that about one-third of motion-sensitive neurons in the pigeon's pretectal nucleus encoded the acceleration of visual motion. Here we propose a microcircuit hypothesis, in which the slow adaptive depressions play a significant role in response generating, to account for the origin of the three important properties of the acceleration-sensitive neurons: the plateau-shaped speed-tuning curves, the opposite-signed after-responses (OSARs) and the acceleration sensitivities. The flat plateau within the speed tuning curves and the OSARs to motion offset observed in experiments are reproduced successfully in simulations, and the simulative responses of the acceleration-sensitive neurons to step changes, ramp changes in stimulus speed and sine wave modulations of stimulus speed are qualitatively consistent with physiological observations. Thus, a biologically plausible substrate for the neurons' classification and the origin of the three properties are provided. PMID- 15791450 TI - AT-rich sequences containing Arabidopsis-type telomere sequence and their chromosomal distribution in Pinus densiflora. AB - Japanese red pine Pinus densiflora has 2 n=24 chromosomes and after FISH detection of Arabidopsis-type (A-type) telomere sequences, many telomere signals were observed on these chromosomes at interstitial and proximal regions in addition to the chromosome ends. These interstitial and proximal signal sites were observed as DAPI-positive bands, suggesting that the interstitial and proximal telomere signal sites are composed of AT-rich highly repetitive sequences. Four DNA clones (PAL810, PAL1114, PAL1539, PAL1742) localized at the interstitial telomere signals were selected from AluI-digested genomic DNA library using colony blot hybridization probed with A-type telomere sequences and characterized using FISH and Southern blot hybridization. The AT-contents of these selected four clones were 60.8-76.3%, and repeat units of the telomere sequence and degenerated telomere sequences were found in their nucleotide sequences. Except for two sites of PAL1114, FISH signals of the four clones co localized with interstitial and proximal A-type telomere sequence signals. FISH signals a showed similar distribution pattern, but the patterns of signal intensity were different among the four clones. PAL810, PAL1539 and PAL 1742 showed similar FISH signal patterns, and the differences were only with respect to the signal intensity of some signal sites. PAL1114 had unique signals that appeared on chromosomes 7 and 10. Based on results of the Southern blot hybridization these four sequences are not arranged tandemly. Our results suggest that the interstitial A-type telomere sequence signal sites were composed of a mixture of several AT-rich repetitive sequences and that these repetitive sequences contained A-type telomere sequences or degenerated A-type telomere sequence repeats. PMID- 15791451 TI - Molecular linkage mapping and phylogeny of the chalcone synthase multigene family in soybean. AB - Chalcone synthase (CHS), the key enzyme in the flavonoid biosynthesis pathway, is encoded by a multigene family, CHS1-CHS8 and dCHS1 in soybean. A tandem repeat of CHS1, CHS3 and CHS4, and dCHS1 that is believed to be located in the vicinity comprises the I locus that suppresses coloration of the seed coat. This study was conducted to determine the location of all CHS members by using PCR-based DNA markers. Primers were constructed based on varietal differences in either the nucleotide sequence of the 5'-upstream region or the first intron of two cultivars, Misuzudaizu, with a yellow seed coat (II), and Moshidou Gong 503, with a brown seed coat (ii). One hundred and fifty recombinant inbred lines that originated from a cross between these two cultivars were used for linkage mapping together with 360 markers. Linkage mapping confirmed that CHS1, CHS3, CHS4, dCHS1, and the I locus are located at the same position in molecular linkage group (MLG) A2. CHS5 was mapped at a distance of 0.3 cM from the gene cluster. CHS2 and CHS6 were located in the middle region of MLGs A1 and K, respectively, while CHS7 and CHS8 were found at the distal end of MLGs D1a and B1, respectively. Phylogenetic analysis indicated that CHS1, CHS3, CHS4, and CHS5 are closely related, suggesting that gene duplication may have occurred repeatedly to form the I locus. In addition, CHS7 and CHS8 located at the distal end and CHS2, CHS6, and CHS members around the I locus located around the middle of the MLG are also related. Ancient tetraploidization and repeated duplication may be responsible for the evolution of the complex genetic loci of the CHS multigene family in soybean. PMID- 15791453 TI - Effects of histone deacetylase inhibitor FR901228 on the expression level of telomerase reverse transcriptase in oral cancer. AB - We speculated whether or not the expression level of telomerase reverse transcriptase (hTERT) would be modulated by agents targeting epigenetics in oral cancer cell lines. Although hTERT is known to be targeted by epigenetic changes, it remains unclear how chemoagents targeting epigenetics work on hTERT transcription. In the present study, the epigenetic effects of the histone deacetylase (HDAC) inhibitor FR901228 on hTERT transcription in oral cancer cell lines were analyzed by RT-PCR. The mRNA expression of hTERT was upregulated after exposure to FR901228 in hTERT-negative Hep2 cells, and even SAS and KB cells expressed high levels of hTERT. Moreover, cotreatment of protein synthesis inhibitor cycloheximide (CHX) resulted in the induction of hTERT transcription by FR901228. This suggests that the induction of hTERT by FR901228 requires de novo protein synthesis to some extent and is more likely a direct than an indirect effect on epigenetic changes such as histone acetylation/deacetylation. We further examined the effect of FR901228 on c-myc protein, which is one of the main hTERT transcription activators. FR901228 repressed c-myc protein only in the absence of CHX, and depended on the enhancement of de novo protein synthesis. Our results indicate that c-myc protein is repressed indirectly by FR901228 but may not contribute to FR901228-induced hTERT transcription. The present study showed that the HDAC inhibitor FR901228 induced the hTERT gene by a complex mechanism that involved transcription factors other than c-myc, in addition to inhibition of histone deacetylation. PMID- 15791452 TI - Analysis of salt-stress-inducible ESTs isolated by PCR-subtraction in salt tolerant rice. AB - To clarify the mechanisms of stress tolerance in rice and to search for rice genes associated with these mechanisms, we analyzed genes induced by a high salinity treatment using the PCR-subtractive hybridization method (PCR subtraction). Seedlings of the salt-tolerant rice cultivar Dee-geo-woo-gen (DGWG) were either treated with 250 mM NaCl for 5 h or left untreated, and PCR subtraction was then performed using the untreated (control) plants as a driver and the NaCl-treated plants as a tester. We obtained 384 clones of tester specific cDNAs as salt-inducible candidates. Northern analysis performed with the cDNA fragments showed that 65 clones had been induced by the NaCl treatment. Sequence analysis and database searching indicated that these clones have homology to proteins functional for detoxification, stress response, and signal transduction in plants. Of these clones, 22% coded for unknown proteins and 12% gave no hits. We selected eight clones from each functional category and analyzed their expression pattern in DGWG. For temporal analysis, seedlings were treated with H(2)O or 250 mM NaCl for 0, 0.5, 1, 2, 5, 10 or 24 h. Different patterns of transcript regulation were found. For the analysis of expression in response to various types of stress and abscisic acid (ABA) treatments, seedlings were treated for 5 h or 10 h with H(2)O, dehydration, cold (4 degrees C), heat (40 degrees C), mannitol, ABA, or wounding. All clones were strongly up-regulated by osmotic stress (dehydration and mannitol) and the ABA treatment. PMID- 15791454 TI - A phase II study of flavopiridol in patients with advanced renal cell carcinoma: results of Southwest Oncology Group Trial 0109. AB - PURPOSE: Flavopiridol is a cyclin-dependent kinase inhibitor that prevents cell cycle progression and tumor growth. In initial phase I studies, encouraging responses were seen in advanced renal cell cancer (RCC). In a phase II study of flavopiridol given as a 72-h continuous infusion every 2 weeks in RCC, a response rate of 6% was seen but with considerable grade 3 or 4 asthenia, diarrhea, and thrombosis. Subsequently, an alternative 1-h bolus schedule was reported to have enhanced tolerability in a phase I trial. We therefore conducted a phase II study of this bolus regimen. METHODS: A total of 38 patients with advanced RCC were entered into this multi-institutional phase II study. Flavopiridol (50 mg/m(2) per day) was administered by bolus intravenous injection daily for three consecutive days, repeated every 3 weeks. RESULTS: Out of 34 eligible patients, one complete response and three partial responses were observed, for an overall response rate of 12% (95% CI 3-27%). Of the 34 patients, 14 (41%) had stable disease (SD). The probability of not failing treatment by 6 months was 21% (95% CI 9-35%). Median overall survival time was 9 months (95% CI 8-18 months). The most common grade 3 or 4 toxicities were diarrhea (35%) and tumor pain (12%) along with anemia, dyspnea, and fatigue (9% each). CONCLUSIONS: Flavopiridol at this dose and schedule is feasible with an acceptable toxicity profile. Flavopiridol has some modest biologic activity against advanced RCC, as evidenced by its single-agent objective response and SD rates. PMID- 15791455 TI - Hyperthermic isolated regional perfusion with CDDP for bone and soft-tissue sarcoma of the lower limb: pharmacokinetics, thermal dose, toxicity, and feasibility. AB - The objectives of this study were to investigate the pharmacokinetics of cisplatin (CDDP) and the thermal dose, toxicity, and feasibility of hyperthermic isolated regional perfusion (HIRP) with CDDP for bone and soft-tissue sarcomas of the lower limb. A total of 43 patients were treated with HIRP using CDDP. The dose of CDDP administered was 62.9+/-11.8 mg/limb (20 mg/m(2) +20 approximately 30 mg). The mean highest CDDP concentration was 56.9 microg/ml as total platinum (tPt) and 49.0 microg/ml as free platinum (fPt). The tPt concentration remained higher than 10 microg/ml. The highest temperature within tumor was 42.3+/-1.4 degrees C on average. The complications of HIRP were grade II toxicity in 30 patients, grade III in 9, and grade IV in 4. The mean necrotic ratio in the resected specimen was 84.5%, and the effect was grade IV (no viable tumor cells) in 13 patients, grade III(>90% necrosis) in 12, grade II (50 to <90%) in 9, and grade I (<50%) in 4. We obtained favorable levels of platinum concentration of the perfusate using a lower CDDP dosage compared with previous studies of HIRP. Considering our results of the pharmacokinetics of CDDP and clinical efficacy, we propose a lower dosage of CDDP for HIRP in the treatment of osteosarcoma. Multimodality treatment of HIRP with preoperative chemotherapy and surgery is a relatively safe and reliable therapeutic option for patients with limb sarcomas, and our method offers excellent local control. PMID- 15791456 TI - American ginseng berry extract and ginsenoside Re attenuate cisplatin-induced kaolin intake in rats. AB - PURPOSE: Cisplatin, a chemotherapeutic agent, causes significant nausea and vomiting. It is postulated that cisplatin-induced oxidant stress may be responsible for these symptoms. We tested whether pretreatment with American ginseng berry extract (AGBE), an herb with potent antioxidant capacity, and one of its active antioxidant constituents, ginsenoside Re, could counter cisplatin induced emesis using a rat pica model. METHODS: In rats, exposure to emetic stimuli such as cisplatin causes significant kaolin intake, a phenomenon called pica. We therefore measured cisplatin-induced kaolin intake as an indicator of the emetic response. Rats were pretreated with vehicle, AGBE (dose range 50-150 mg/kg, IP) or ginsenoside Re (2 and 5 mg/kg, IP). Rats were treated with cisplatin (3 mg/kg, IP) 30 min later. Kaolin intake, food intake, and body weight were measured every 24 h for 120 h. Additionally, the free radical scavenging activity of AGBE was measured in vitro using ESR spectroscopy. RESULTS: A significant dose-response relationship was observed between increasing doses of pretreatment with AGBE and reduction in cisplatin-induced pica. Kaolin intake was maximally attenuated by AGBE at a dose of 100 mg/kg. Food intake also improved significantly at this dose (P<0.05). Pretreatment with ginsenoside Re (5 mg/kg) also decreased kaolin intake (P<0.05). In vitro studies demonstrated a concentration-response relationship between AGBE and its ability to scavenge superoxide and hydroxyl radicals. CONCLUSION: Pretreatment with AGBE and its major constituent, Re, attenuated cisplatin-induced pica, and demonstrated potential for the treatment of chemotherapy-induced nausea and vomiting. Significant recovery of food intake further strengthens the conclusion that AGBE may exert an antinausea/antiemetic effect. PMID- 15791457 TI - Targeting BCL-2 overexpression in various human malignancies through NF-kappaB inhibition by the proteasome inhibitor bortezomib. AB - BACKGROUND: BCL-2 overexpression occurs in many cancer types and is associated with chemoresistance and radioresistance. The mechanisms responsible for its aberrant expression are thought to be transcriptionally mediated but remain unclear. We examined the cell type-specific mechanism of BCL-2 gene transcription in various solid organ malignancies. METHODS: Regulation of BCL-2 gene transcription was examined in seven different human cancer cell lines including two pancreatic (MIA-PaCa-2, PANC-1), two prostate (LNCaP, PC-3), two lung (Calu 1, A549) and one breast (MCF-7) cancer cell line. Cells were treated with inhibitors of phosphatidylinositol-3 kinase (PI3K), MEK/ERK, and p38MAPK. The effect of mutation of a NF-kappaB site in the BCL-2 promoter was determined, as was the effect of inhibition of NF-kappaB function using a 26S proteasome inhibitor (bortezomib) on both BCL-2 transcription and induction of apoptosis. RESULTS: BCL-2 expression varied both between and within tumor types; four of seven cell lines demonstrated high BCL-2 levels (MIA-PaCa-2, PC-3, Calu-1 and MCF 7). No signaling pathway was uniformly responsible for overexpression of BCL-2; however, mutation of the NF-kappaB site decreased BCL-2 promoter activity in all cell lines. Inhibition of NF-kappaB activity decreased BCL-2 protein levels independently of the signaling pathway involved in transcriptional activation of the BCL-2 gene. CONCLUSIONS: Diverse signaling pathways variably regulate BCL-2 gene expression in a cell type-specific fashion. Therapy to decrease BCL-2 levels in various human cancers would be more broadly applicable if targeted to transcriptional activation rather than signal transduction cascades. Finally, the apoptotic efficacy of proteasome inhibition with bortezomib paralleled the ability to inhibit NF-kappaB activity and decrease BCL-2 levels. PMID- 15791458 TI - In vivo antitumor efficacy of 17-DMAG (17-dimethylaminoethylamino-17 demethoxygeldanamycin hydrochloride), a water-soluble geldanamycin derivative. AB - PURPOSE: To describe the preclinical basis for further development of 17-dimethyl aminoethylamino-17-demethoxygeldanamycin hydrochloride (17-DMAG, NSC 707545). METHODS: In vitro proliferation assays, and in vivo model studies in metastatic pancreatic carcinoma and subcutaneous xenograft melanoma and small-cell lung carcinoma models. RESULTS: 17-DMAG emerged from screening studies as a potent geldanamycin analog, with the average concentration inhibiting the growth of the NCI anticancer cell line drug screen by 50% being 0.053 microM. "Head to head" comparison with 17-allylamino-17-demethoxygeldanamycin (17-AAG, NSC 330507) revealed 17-DMAG to possess potent activity against certain cell types, e.g., MDA MB-231 breast carcinoma and HL60-TB leukemia which were relatively insensitive to 17-AAG. Evidence of oral bioavailability of 17-DMAG in a saline-based formulation prompted more detailed examination of its antitumor efficacy in vivo. 17-DMAG inhibited the growth of the AsPC-1 pancreatic carcinoma xenografts growing as intrahepatic metastases at doses of 6.7-10 mg/kg twice daily for 5 days administered orally under conditions where 17-AAG was without activity. 17-DMAG in an aqueous vehicle at 7.5-15 mg/kg per day for 3 days on days 1-3, 8-10 and 13 17, or 1-5 and 8-12 showed evidence of antitumor activity by the parenteral and oral routes in the MEXF 276 and MEXF 989 melanomas and by the parenteral route in the LXFA 629 and LXFS 650 adenocarcinoma and small-cell carcinoma models. The latter activity was comparable to the historical activity of 17-AAG. CONCLUSIONS: Taken together, the in vivo activity of 17-DMAG supports the further development of this water-soluble and potentially orally administrable geldanamycin congener. PMID- 15791459 TI - Properties of recombinant human N1-acetylpolyamine oxidase (hPAO): potential role in determining drug sensitivity. AB - The recent cloning of the mammalian gene coding for N(1)-acetylpolyamine oxidase (PAO) provides the opportunity to directly examine the role of human PAO (hPAO) in polyamine homeostasis as well as its potential role in determining cellular response to antitumor polyamine analogues. To facilitate the study of this enzyme, the production, purification, and characterization of the recombinant hPAO is reported. hPAO oxidizes N(1)-acetylspermidine (K(m)=2.1 microM, K(cat)=15.0 s(-1)) and has very high affinity for N(1)-acetylspermine (K(m)=0.85 microM, K(cat)=31.7 s(-1)). The recombinant hPAO does not efficiently oxidize spermine, thereby demonstrating a significant difference in substrate specificity from the previously described human spermine oxidase PAOh1/SMO. Importantly, hPAO demonstrates the ability to oxidize a subset of antitumor polyamine analogues, suggesting that this oxidase activity could have a significant effect on determining tumor sensitivity to these or similar agents. Transfection of A549 human lung cancer cells with an hPAO-expressing plasmid leads to a profound decrease in sensitivity to those analogues which act as substrates, confirming its potential to alter drug response. One similarity that hPAO shares with human PAOh1/SMO, is that certain oligoamine analogues are potent inhibitors of its oxidase activity. The results of these studies demonstrate how changes in polyamine catabolism may affect drug response. PMID- 15791461 TI - Incidence and risk factors for paclitaxel hypersensitivity during ovarian cancer chemotherapy. AB - Hypersensitivity reaction (HSR) is still a major concern during cancer chemotherapy with paclitaxel. In the present study, we investigated retrospectively the incidence of HSRs to paclitaxel and the risk factors in 105 patients (553 courses) who received adjuvant chemotherapy (paclitaxel and carboplatin) for ovarian cancer. Moderate to severe HSRs that led to cessation or discontinuation of the chemotherapy, including respiratory distress and hypotension, were observed in 14 patients (13.3%) and 16 courses (2.9%), regardless of the use of conventional premedication with glucocorticoid, and histamine H(1) and H(2) antagonists. The incidence of HSRs to paclitaxel in patients with ovarian cancer seemed to be considerably higher than those reported by other investigators in patients with other carcinomas such as non-small-cell lung cancer and breast cancer. Four risk factors were identified: (1) history of mild dermal reactions such as facial flushing and urticaria in previous courses, (2) presence of respiratory dysfunction, (3) obesity (body mass index >25), and (4) postmenopausal at the time of ovariectomy. The incidence of hypersensitivity increased linearly as the number of risk factors increased (r=0.992, P=0.008). It is likely that disappearance of the estrous cycle facilitates the occurrence of HSRs to paclitaxel. PMID- 15791460 TI - Opioid growth factor enhances tumor growth inhibition and increases the survival of paclitaxel-treated mice with squamous cell carcinoma of the head and neck. AB - Paclitaxel is used as a single agent, and in combination with other drugs, as a standard of care in the treatment of squamous cell carcinoma of the head and neck (SCCHN). However, the use of paclitaxel for therapy of SCCHN may be accompanied by serious side effects. Paclitaxel is a known cytotoxic inhibitor of cell proliferation that acts by stabilizing microtubules and inducing apoptosis. Opioid growth factor (OGF), [Met(5)]-enkephalin, is an endogenous peptide that has tonically active inhibitory effects on the growth of SCCHN in vitro and in vivo. OGF action is rapid, reversible, mediated by the nuclear-associated OGF receptor (OGFr), and is not cytotoxic (nor apoptotic related). The present study was designed to examine whether a combination of chemotherapy with paclitaxel and biotherapy with OGF is more effective than either agent alone in inhibiting tumor growth. Moreover, focus was placed on whether there are changes in the side effects known to occur with paclitaxel alone, following this combined therapy. Human SCC-1 cells, derived from a well differentiated SCCHN, were transplanted into athymic mice. The mice were randomized to receive intraperitoneal (i.p.) injections of sterile saline (controls), OGF (10 mg/kg, daily), paclitaxel (8 mg/kg, every other day), or both paclitaxel (8 mg/kg, every other day) and OGF (10 mg/kg, daily) beginning on the day of tumor inoculation. OGF, but not paclitaxel, delayed measurable and visible tumor appearance of mice with SCCHN. Treatment with paclitaxel, but not with other agents, had a marked effect on the body weights. Survival only was reduced in the paclitaxel group, with an average life span of 34.3+/-3.1 days recorded, in comparison to the 50-day survival (date of termination) for all other groups. Beginning after week 4 of tumor inoculation and drug treatment, the tumor weight of the paclitaxel/OGF group was significantly reduced from the control, OGF, and paclitaxel-exposed mice. The OGFr number of the SCCHN tumors was 2.1-fold greater in the animals exposed to OGF or paclitaxel, and elevated 38% in the paclitaxel/OGF group; significant differences from the control group were found for the OGF and paclitaxel groups. These data suggest that combined chemotherapy (i.e., paclitaxel) and biotherapy (OGF) provides a valuable alternative to the standard of care for SCCHN patients. PMID- 15791462 TI - Localization of the human female breast in primary somatosensory cortex. AB - RATIONALE: Despite an extensive body of research on the topography of the primary somatosensory cortex (S1) little is known about the representation of the trunk. AIM: The aim of this study was to determine the representation of the breast in S1 in human females. RESULTS: The representation of the human breast in primary somatosensory cortex was determined in ten healthy female subjects. Non-painful electrical stimulation of the mammilla (Th4 dermatome), groin (L1 dermatome) and the first digit of both sides of the body activated cutaneous receptors and thus elicited somatosensory evoked potentials. The representation of these body parts in primary somatosensory cortex (S1) was determined using neuroelectric source imaging. Equivalent current dipole localizations were overlaid with individual structural magnetic resonance images to account for individual cortical differences. The breast representation was localized between the representation of the groin and the first digit. In the medial-lateral direction the representation of the breast was approximately 15 mm lateral of the longitudinal fissure in the contralateral hemisphere. Source localizations were stable across subjects. However, one subject showed ipsilateral representation of the breast, which might be related to bilateral receptive fields of the ventral body midline representation. This study confirms the Penfield and Rasmussen (1950) invasive data by use of noninvasive source imaging. PMID- 15791463 TI - How does action resist visual illusion? Uncorrected oculomotor information does not account for accurate pointing in peripersonal space. AB - Using spatially identical displays (variants of the Muller-Lyer illusion), we compared the accuracy of spatial verbal judgments with that of saccadic (eye) and pointing (hand) movements. Verbal judgments showed a clear effect of the illusion. The amplitude of the primary saccade from one endpoint of the pattern (at fixation) to the other also showed an effect of the illusion. Conversely, movement amplitudes when pointing from one endpoint (initial finger position) to the other were significantly more accurate than both saccades and verbal responses. In a control experiment in which the viewing conditions between the saccade and pointing experiments were equalized, saccade amplitude was again affected by the illusion. In several studies, systematic biases in conscious spatial judgments have been contrasted with accurate open-loop pointing in peripersonal space. It has been proposed that such seeming dissociations between vision-for-action and vision-for-consciousness might in fact be because of a simple oculomotor strategy: saccade to the target before it disappears, then use the efference copy of the (accurate) saccadic movement to drive pointing. The present data do not support the hypothesis in this simple form. PMID- 15791464 TI - Visual fixation of a landing perch by chickens. AB - Chickens were video recorded while making jumps or flights toward a landing perch, to test hypotheses about visual fixation behaviour. In the first experiment, varying the height above the landing perch of the food container providing the incentive to jump had no effect on head orientation, indicating that the birds were not fixating this object. In the second experiment, hens jumped over six combinations of perch height and distance, and a linear relationship was found at take-off between head orientation and the angular distance of the perch from the horizontal at the eye. This relationship is consistent with fixation of the perch by a linear combination of head and eye rotations, with the head component contributing 73% of the total response. The image of the perch is fixated 20 masculine below that of the bill tip, outside any region of the chicken retina specialised for high acuity vision. Fixation of the perch before jumping must therefore have some function other than inspection with high acuity, such as providing a constraint that enables precise visual control of trajectory and landing manoeuvres. PMID- 15791465 TI - Feeling what you hear: auditory signals can modulate tactile tap perception. AB - We tested whether auditory sequences of beeps can modulate the tactile perception of sequences of taps (two to four taps per sequence) delivered to the index fingertip. In the first experiment, the auditory and tactile sequences were presented simultaneously. The number of beeps delivered in the auditory sequence were either the same as, less than, or more than the number of taps of the simultaneously presented tactile sequence. Though task-irrelevant (subjects were instructed to focus on the tactile stimuli), the auditory stimuli systematically modulated subjects' tactile perception; in other words subjects' responses depended significantly on the number of delivered beeps. Such modulation only occurred when the auditory and tactile stimuli were similar enough. In the second experiment, we tested whether the automatic auditory-tactile integration depends on simultaneity or whether a bias can be evoked when the auditory and tactile sequence are presented in temporal asynchrony. Audition significantly modulated tactile perception when the stimuli were presented simultaneously but this effect gradually disappeared when a temporal asynchrony was introduced between auditory and tactile stimuli. These results show that when provided with auditory and tactile sensory signals that are likely to be generated by the same stimulus, the central nervous system (CNS) tends to automatically integrate these signals. PMID- 15791466 TI - Exteroceptive reflexes in jaw-closing muscle EMG during rhythmic jaw closing and clenching in man. AB - Exteroceptive jaw reflexes might play a role in normal functions of the mouth such as mastication. Until now these reflexes have only been studied under isometric conditions. The aim of this study was to compare exteroceptive reflexes in jaw muscle EMG during the closing phase of rhythmic open-close movements and clenching, at the same jaw gape and with similar muscle EMG. Reflexes consisting of successive waves of decreased and increased muscle activity (the Q, R, S and T waves of the post-stimulus electromyographic complex (PSEC)), evoked by light noxious electrical stimulation of the vermillion border of the lower lip, were recorded from the jaw closing muscles of 17 subjects. Differences between the two tasks occurred in two phases of the PSEC: (1) in an early phase, around the R wave, there was significantly less EMG during jaw closing (mean EMG ratio between jaw-closing and clenching 0.71), and (2) in a late phase, around the transition between the S to the T wave, there was significantly more EMG during jaw closing (mean EMG ratio: 1.40). The decrease in EMG activity around the R wave during jaw closing may be due to a change in reflex sensitivity at an interneuron level. The increase in EMG activity around the transition between the S and T waves during jaw closing might, at least in part, be due to a proprioceptive stretch reflex. This reflex is mediated by muscles spindles that are activated by the deceleration of the jaw evoked by the lip stimulus. The finding of inhibitory reflex mechanisms that predominate more during rhythmic jaw movements than during clenching in an early phase of the PSEC might be related to protecting oral tissues from trauma when the jaw is closing with potentially a large muscle force. In contrast, when food is held between the teeth, a possible inhibitory influence of light noxious stimuli is diminished. PMID- 15791467 TI - Inhibition of the growth of urinary calcium hydrogen phosphate dihydrate crystals with aqueous extracts of Tribulus terrestris and Bergenia ligulata. AB - Urinary type calcium hydrogen phosphate dihydrate (CHPD) or Brushite crystals were grown by the single diffusion gel technique in silica hydro-gels. The gel framework acts as a three dimensional crucible in which the crystal nuclei are delicately held in the position of their formation and nutrients are supplied for their growth. This technique can be utilized as a simplified screening model to study the growth and dissolution of urinary stones in vitro. The action of the putatively litholytic medicinal plants Tribulus terrestris and Bergenia ligulata on the growth of CHPD crystals was studied . The effects of artificial reference urine (ARU) and human urine (HU), along with the plant extracts, are also reported. Attempts were made to understand the role of these inhibitors on urinary crystal formation. HU, ARU, extracts of B. ligulata and T. terrestris exhibit appreciable amounts of inhibition, but B.ligulata and T.terrestris with ARU and HU do not show inhibition at all. PMID- 15791469 TI - Combination of surgery and immunotherapy in metastatic renal cell carcinoma. AB - The treatment of choice for non-disseminated renal cell cancer (RCC) is surgery. However, the 5-year survival rates for all stages do not exceed 60%, even in contemporary series. Further improvement will most likely have to await the development of a more effective systemic therapy and the application of combined treatment modalities to counter the relatively high number of patients presenting with advanced stages. Whereas textbook belief up to the 1990s suggested refraining from surgical antitumor-therapy in the case of metastatic RCC, current strategies clearly advocate debulking tumor nephrectomy in the context of modern immunotherapies. This dramatic change of attitude stemmed from two randomized phase III trials conducted by EORTC and SWOG, including a combined analysis of both studies, in which cytoreductive tumor nephrectomy conveyed a significant survival benefit over immunotherapy alone. Concepts and progress in this field appear to be of major interest for modern oncologic urologists following the advent of immunotherapeutic strategies that require surgical intervention at some stage of the treatment cascade. PMID- 15791468 TI - Treatment of brain metastases in renal cell carcinoma: radiotherapy, radiosurgery, or surgery? AB - Metastases from renal cell carcinoma raise specific therapeutic problems because they are relatively unresponsive to whole brain radiation therapy and tend to bleed. Recently, stereotactically guided high-precision irradiation as a single dose application (radiosurgery) showed promising treatment results for selected patients with brain metastases from renal cell carcinoma. Radiosurgery appears attractive due to its low risk and minimal invasiveness. Multiple lesions can be treated at the same time and retreatments can be performed for local or distant recurrences. PMID- 15791471 TI - Immobilization as a technical possibility for long-term storage of bacterial biosensors. AB - For applications in field experiments, the recombinant strain Salmonella typhimurium TA1535 was immobilized to permit its immediate utilization after long storage periods. Salmonella typhimurium TA1535 cells contain the plasmid that has an inducible SOS promoter fused to a promoterless luxCDABFE operon from Photobacterium leiognathi. The induction of bioluminescence occurs in the presence of the DNA-damaging agent mitomycin C which stimulates the bacterial SOS response. Early stationary phase cells were immobilized at a cell concentration of 10(10) CFU/ml in microtiter plates and stored up to 6 weeks at 4 degrees C in a sealed container. Even after 4 weeks of storage, the bioluminescence kinetics and yield in response to different concentrations of mitomycin C were not significantly different from those of freshly prepared samples. PMID- 15791472 TI - Physiological parameters that affect the transfer of radiocaesium to ruminants. AB - Recently there has been a renewed interest in biological scaling relationships between parameters, such as those between, for example, body mass, dry matter intake and biological half-times of radionuclides that are useful in predicting the transfer of radiocaesium to different animal species, particularly to wild animals. However, there is still a considerable unexplained variability in transfer coefficient estimates between individuals of the same species. This paper discusses the physiological parameters that affect the transfer of radiocaesium to ruminants, and it shows how a better understanding of these parameters may help to reduce the within-species variability in radiocaesium transfer coefficients. In light of the improved understanding during the past 10 15 years of the importance of source-dependent bioavailability on absorption of radiocaesium from the gastrointestinal tract, it is concluded that further studies are required on the effects of feed digestibility and physiological factors on absorption and endogenous faecal excretion of radiocaesium to better understand the variability in transfer coefficients. PMID- 15791470 TI - Skull vault growth in craniosynostosis. AB - BACKGROUND: Since its first description by Virchow, the principle of abnormal skull growth due to restriction of skull growth at the fused sutures, and the realisation by Moss that the sutures at the skull base are equally affected, have been the main intellectual driving forces behind the majority of cranial expansion procedures performed currently in children with craniosynostosis. CURRENT OBSERVATIONS: Despite original impressions that craniosynostosis leads to craniostenosis, many studies have demonstrated that in the majority of patients with craniosynostosis there is normal skull volume in those over the age of 6 months. In Apert syndrome, skull volume is invariably larger than normal. Some studies have shown that intracranial pressure is independent of intracranial volume, and can exist in the presence of normal volume, or indeed after cranial expansion. These observations imply that cranial expansion procedures create a state of artificially increased skull volume, in the quest to improve appearance and function. FUTURE ADVANCES: This creates a new angle of view through which skull growth abnormalities are seen. It is becoming clearer that in most patients with craniosynostosis, there is regional imbalance of skull growth, which co exists with a variety of other equally important factors, such as genetic defects, raised intracranial pressure, venous hypertension, and other brain parenchymal anomalies such as hindbrain hernia or hydrocephalus. It is becoming increasingly obvious that the type of surgical treatment currently practised in most cases is conceptually incorrect. Recent modifications such as the use of springs or distraction do not escape from the underlying philosophy of cranial expansion. With that in mind, it is hoped that advances in the fields of genetics and molecular biology will provide treatments for the cause of craniosynostosis rather than the symptomatic relief that surgery offers currently. CONCLUSION: Until then, there is a need to develop better ways of quantifying regional abnormalities of skull growth. PMID- 15791473 TI - Health-related and work-related aspects associated with sick leave: a comparison of chronically ill and non-chronically ill workers. AB - OBJECTIVES: Our aims were to study to what extent chronically ill workers (CIWs) take more sick leave than non-chronically ill workers (NCIWs) and to explore which health-related and work-related aspects are associated with the sick leave patterns of the two groups. METHODS: A questionnaire on work, health and sick leave was sent to all employees of a university in The Netherlands (response: 49.1%). Analyses were conducted for 444 CIWs and 1,347 NCIWs. Odds ratios (ORs) were calculated to quantify the contribution of being chronically ill to sick leave in general, frequent sick leave, prolonged sick leave, and present sick leave. The contributions of health-related and work-related aspects to sick leave were investigated by multiple logistic regression analyses for both CIWs and NCIWs separately. RESULTS: CIWs showed significantly increased ORs for general, frequent, prolonged and present sick leave when compared with NCIWs. Fatigue, emotional exhaustion and perceived health complaints showed stronger associations with sick leave for both CIWs and NCIWs than various work-related aspects. Workers of 46 years of age and older showed less sick leave than workers under the age of 36. Male respondents and scientific personnel showed less frequent sick leave than the other respondents, and so did respondents working more than 40 h a week, compared with part-timers. The final regression models explained 8% 16% of the variance in sick leave. CONCLUSIONS: CIWs take two to three times more and longer sick leave than NCIWs. Health-related aspects are more strongly associated with sick leave than work-related aspects for both CIWs and NCIWs. Sick leave patterns were, nevertheless, only partly explained by health-related and work-related aspects. In any case, future studies of sick leave should certainly take the presence of chronic disease into account as an important determinant of sick leave. PMID- 15791475 TI - Cervical adult rhabdomyoma presenting as a rapidly growing mass in a patient with diffuse large B-cell non-Hodgkin's lymphoma. AB - BACKGROUND: Adult rhabdomyoma is a rare mesenchymal tumor, which generally grows slowly and is mainly localized in the head and neck area. PATIENT AND METHODS: We report the extraordinary case of a rapidly growing adult rhabdomyoma in a 73-year old man. The patient was treated for diffuse large B-cell non-Hodgkin's lymphoma with CHOP therapy (doxorubicin, cyclophosphamide, vincristine, and prednisone). Comparison of the respective computed tomography scans showed prominent enlargement of 35% in the tumor mass volume on the right side of the neck within 3 months. The tumor was highly suspicious for lymphoma. Surgical resection was performed. RESULTS: Histological examination revealed a tumor which was composed of tightly packed polygonal cells with a PAS-positive granular or vacuolated cytoplasm, occasionally with cross-striations. Immunohistochemically, the cells were positive for desmin, myogenin, Myo-D1, but negative for S-100. Due to these characteristic morphologies, adult rhabdomyoma was diagnosed. CONCLUSION: This is the first report on an adult rhabdomyoma with a proven rapid enlargement. The possible pathomechanisms are discussed. PMID- 15791476 TI - Spatial competition for quality in the market for hospital care. AB - This study analyses the market for secondary health care services when patient choice depends on the quality/distance mix that achieves utility maximization. First, the hospital's equilibrium in a Hotelling spatial competition model under simultaneous quality choices is analyzed to define hospitals' strategic behavior. A first equilibrium outcome is provided, the understanding of which is extremely useful for the policy maker wishing to improve social welfare. Second, patients are assumed to be unable, because of asymmetry of information, to observe the true quality provided. Their decisions reflect the perceived quality, which is affected by bias. Using the mean-variance method, the equilibrium previously found is investigated in a stochastic framework. PMID- 15791474 TI - Work-related disorders of the upper limb in female workers using orbital sanders. AB - OBJECTIVES: To investigate the prevalence of vascular, neurological and musculoskeletal disorders of the upper limb in a group of female workers performing either mechanical or hand sanding in the furniture industry (n=100) and in a control group of female office workers (n=100). A further aim was to study the possible adverse effects of exposure to a combination of vibration and ergonomic risk factors in female workers. METHODS: All female workers underwent a medical interview and a complete physical investigation. The clinical diagnoses of Raynaud's phenomenon and carpal tunnel syndrome (CTS) were made according to international consensus criteria. Hand-transmitted vibration from orbital sanders was measured according to International Organization for Standardization (ISO) recommendations. Daily vibration exposure was assessed in terms of 8-h energy equivalent frequency-weighted acceleration [A(8)] according to the European Directive on physical agents. Ergonomic load on the upper limbs was estimated by means of strain index methodology. RESULTS: There was no significant difference in the prevalence of Raynaud's phenomenon between the furniture workers (4%) and the controls (8%). The prevalence of CTS, peripheral sensorineural disturbances (after exclusion of CTS cases) and upper-limb musculoskeletal complaints was significantly greater in the furniture workers than in the controls. CTS was clinically diagnosed in 19% of the furniture workers and 8% of the controls. A log-binomial regression analysis showed that the occurrence of soft-tissue disorders of the upper limb increased significantly with the increase of both daily vibration exposure and the strain index score. It was estimated that the risk for CTS increased by a factor of 1.30 (95% CI 1.11-1.53) for each unit of increase in A(8) (ms(-2)), and by 1.09 (95% CI 1.02-1.15) for each unit of increase in the strain index score. CONCLUSIONS: Even though the cross-sectional design of this epidemiological study does not allow aetiological considerations, our findings suggest a significant association between occupational use of vibratory tools and soft-tissue disorders in the upper limbs of female workers. Quantitative estimation of vibration exposure and ergonomic stress showed that these physical risk factors seem to contribute in a multiplicative way to the occurrence of chronic nerve and musculoskeletal disorders in female workers operating hand-held vibrating tools in the furniture industry. PMID- 15791477 TI - Prolonged inhibition of nitric oxide synthesis in pregnant rats: effects on blood pressure, fetal growth and litter size. AB - METHODS: In order to investigate the effect of chronic inhibition of nitric oxide synthesis along pregnancy, pregnant rats were given drinking water alone (control group) or drinking water containing nonselective nitric oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME; 15 mg/day/rat equivalent to 50 mg/kg/day; treated group), from postmating days 1 to 18 of pregnancy. On days 1 to 17 of pregnancy, urinary volume, urinary sodium and potassium excretion, and urine protein concentration were measured. Systolic blood pressure (SBP) was recorded daily. On days 6, 11 and 18 of pregnancy the number of sites of implantation, number of embryos, litter size, fetal and placenta weight were determined. RESULTS: Systolic blood pressure (mmHg) increased (p<0.001) on the 2nd day of administration of L-NAME and remained high throughout the experiment. This treatment increased urinary protein excretion and urine volume (p<0.01), with changes in the sodium and potassium excretion rate along the study. On day 6 of gestation in treated group, the number of implantation sites (0.14+/-0.10) significantly decreased (p<0.05) compared with the control group (1.45+/-0.58), but on day 11 of pregnancy the number of embryos was similar in both groups. By day 18 of pregnancy, L-NAME caused a substantial decrease (p<0.001) in litter weight (6.30+/-0.77 to 12.00+/-0.92 g), weight of placenta (3.17+/-0.22 to 4.74+/ 0.21 g) (p<0.001) and litter size (7.95+/-0.59 to 11.95+/-0.45 fetus/litter; p<0.001). Also, treatment with L-NAME caused an important number of fetal resorptions (2.93+/-0.42 No./litter to 0 in control group). CONCLUSION: Thus, treatment of pregnant rats with L-NAME, has an important effect on systolic blood pressure and on the physiology of reproduction, mainly in the third stage of pregnancy. PMID- 15791478 TI - Placenta percreta invading the urinary bladder. AB - INTRODUCTION: Placenta percreta is a rare obstetric complication causing life threatening hemorrhage. CASE REPORT: The case of a woman with a placenta percreta invading the urinary bladder treated by cesarean hysterectomy and partial bladder resection is presented. Overall estimated blood loss was 11,130 ml, and 59 units of various blood products were transfused. CONCLUSION: Obstetricians and urologists should be aware of this rare condition. PMID- 15791480 TI - Tumour necrosis factor-mediated cell death pathways do not contribute to muscle fibre death in dystrophinopathies. AB - There is evidence that apoptotic cell death mechanisms contribute to muscle fibre loss in dystrophinopathies, but little knowledge about the activators of the final degrading caspase cascade in muscle fibre apoptosis. As mitochondria related activation of this caspase cascade, through e.g. APAF-1, could not be proven in dystrophin-deficient muscle, this study searches for other prospective candidates that may directly trigger apoptotic cell degradation by mitochondria independent pathways involving the interaction of tumour necrosis factor-alpha (TNF-alpha) and TRAIL with death receptors and subsequent activation of caspase 8. The expression of TNF-alpha, TNF-R1, TRAIL, NF-(kappa)B and caspase-8 were studied in muscle biopsy specimens from 14 patients with a dystrophinopathy [10 Duchenne muscular dystrophies (DMD), 2 Becker MD, and 2 DMD carriers] by immunohistochemistry and Western blotting. In all types of dystrophinopathies, necrotic muscle fibres undergoing myophagocytosis displayed strong expression of TNF-alpha, TNF-R1, and TRAIL, which, however, was attributed to phagocytosing cells and not to the muscle fibres themselves. There was no up-regulation in normal-shaped or atrophic non-necrotic muscle fibres, or in intact muscle fibre segments adjacent to segmental necrosis and myophagocytosis. The expression profiles of caspase-8 and NF-(kappa)B resembled that of normal control muscle. There were likewise no significant differences in the Western blot analyses between normal control and dystrophin-deficient muscle. Based on these findings, a contribution of TNF-alpha or TRAIL-mediated cell death pathways to muscle fibre apoptosis or necrosis in dystrophinopathies could not be confirmed. PMID- 15791479 TI - Mutation analysis of B-RAF gene in human gliomas. AB - The RAS/RAF/MEK/ERK kinase pathway is pivotal in the transduction of mitogenic stimuli from activated growth factor receptors, which regulates cell proliferation, survival, and differentiation. Up-regulation of this pathway due to RAS mutations is found in approximately 30% of human tumors. Recently, activating mutations of B-RAF were identified in a large proportion of human cancers. Gliomas are the most frequent primary central nervous system tumors and the molecular mechanisms that underlie the development and progression of these tumors are far from being completely understood. The purpose of this study was to clarify the incidence of B-RAF mutations and their possible relation with tumor progression in a series of 82 human gliomas, including 49 astrocytic and 33 oligodendroglial tumors. The analysis of B-RAF hotspot regions, exons 11 and 15, showed presence of B-RAF mutations in only 2 out of 34 (6%) glioblastomas, and absence in the remaining histological types. Both mutations were located in the hotspot residue 600 (V600E) at exon 15, which leads to constitutive B-RAF kinase activity. These data suggest that activating mutations of B-RAF are not a frequent event in gliomas; nevertheless, when present they are associated with high-grade malignant lesions. PMID- 15791481 TI - Acute hemorrhagic leukoencephalitis (Hurst's disease) linked to Epstein-Barr virus infection. AB - A 16-year-old girl presented signs of a common cold in combination with a hemolytic crisis. Within 3 days, she developed reduced consciousness and hemiparesis subsequently followed by coma. CT and MRI scans revealed evidence for raised intracranial pressure and an extensive inflammatory process extending from the brain stem up to the thalamus. The patient died within 3 weeks after onset of first symptoms of intracranial pressure despite maximum intensive care. Neuropathological examination revealed disseminated necrotic lesions and perivascular hemorrhages characteristic for acute hemorrhagic leukoencephalitis (Hurst's disease), mainly of the brain stem, diencephalon and cerebellum. Serological results, in situ hybridization and PCR analysis demonstrated an acute Epstein-Barr virus (EBV) infection of the central nervous system. To our knowledge, this is the first reported case of Hurst's disease linked to EBV. PMID- 15791482 TI - Heterogeneity in distribution of amyloid-positive islets in type-2 diabetic patients. AB - Amyloid-containing (A+) islets are characteristic for type-2 diabetes (T2D), but their abundance seems variable among patients. It is unclear whether the distribution of A+ islets follows a certain pattern or occurs randomly throughout the pancreatic organ. We investigated the topography of A+ islets in eight pancreata of T2D patients and eight sex- and age-matched non-diabetic subjects. Transversal sections of head, body and tail segments were stained with synaptophysin combined with Congo red to map/quantify islet tissue and amyloid. In the eight T2D pancreata, the overall percentage of A+ islets varied from 4% to 85%. Further analysis in body and tail indicated that peripheral regions exhibited higher percentages of A+ islets than central regions (averages of, respectively, 30% and 17%, P<0.05). Non-diabetic control pancreata also exhibited A+ islets, albeit at a 25-fold lower frequency; a tendency towards higher percentage of A+ islets in peripheral versus central regions was also observed. The higher percentage A+ islets in peripheral regions was associated with a higher density and relative islet over exocrine surface area. These observations on heterogeneity in abundance and distribution of A+ islets need consideration when sampling tissue for studies on human islet amyloidosis. The present methodology allows us to further investigate the susceptibility to amyloidosis of islets in peripheral regions of the pancreas. PMID- 15791483 TI - Hydrogen peroxide sensor based on horseradish peroxidase immobilized on a silver nanoparticles/cysteamine/gold electrode. AB - A third-generation hydrogen peroxide biosensor was prepared by immobilizing horseradish peroxidase (HRP) on a gold electrode modified with silver nanoparticles. A freshly-cleaned gold electrode was first immersed in a cysteamine-ethanol solution, and then silver nanoparticles were immobilized on the cysteamine monolayer, and finally HRP was adsorbed onto the surfaces of the silver nanoparticles. This self-assemble process was examined via atomic force microscopy (AFM). The immobilized horseradish peroxidase exhibited an excellent electrocatalytic response toward the reduction of hydrogen peroxide. The linear range of the biosensor was 3.3 microM to 9.4 mM, and the detection limit was estimated to be 0.78 microM. Moreover, the biosensor exhibited a fast response, high sensitivity, good reproducibility, and long-term stability. PMID- 15791484 TI - A multiresidue method using ion-trap gas chromatography-tandem mass spectrometry with or without derivatisation with pentafluorobenzylbromide for the analysis of pesticides in the atmosphere. AB - A multiresidue method using gas chromatography coupled to ion-trap tandem mass spectrometry (MS/MS) was developed for the analysis of 27 pesticides, commonly used in Alsace, in atmospheric samples (particle and gas phases). As pesticides are expected to be present at very low concentrations and in a complex matrix, the analytical method used was both highly selective and sensitive. These two properties were obtained by associating chromatography with ion-trap MS/MS. To develop this method, analysis of electron impact in single MS was first conducted to choose the parent ions of the pesticides studied. Among the 27 pesticides analysed, seven of them require a derivatisation step. This was the case of some ureas (chlorotoluron, diuron and isoproturon), phenoxy acids (2,4 dichlorophenoxyacetic acid, 4-chloro-2-methylphenoxyacetic acid and mecoprop) and of bromoxynil. The derivatisation was performed with success with pentafluorobenzylbromide. Then, a MS/MS method was optimised by parameters such as the radio frequency storage level and the collision-induced dissociation excitation voltage. Finally, a last step enabled the development of two calibrating programs based on the quantification of daughter ions for the 20 pesticides analysed directly (run 1) and for the seven pesticides which needed derivatisation (run 2). With this analytical procedure, the detection limits varied between 2.5 and 1,250 pg m(-3) depending on the compounds studied. This method was tested with success for atmospheric samples collected in Strasbourg (France) during intensive pesticide treatment in 2002. PMID- 15791485 TI - Mesenteric inflammatory myofibroblastic tumor mimicking a necrotized malignant mass in an adult: case report with MR findings. AB - Inflammatory myofibroblastic tumors are lesions that usually affect young adults and children. Numerous extrapulmonary sites of these tumors have been found, but the mesentery is a very unusual location for an inflammatory myofibroblastic tumor and it is extremely rare in an older patient. Inflammatory myofibroblastic tumor of the mesentery is often demonstrated as a large and infiltrative lesion with heterogeneous enhancement. Its diagnosis is recognized as difficult and it often mimics malignant tumor. We report a case of mesenteric inflammatory myofibroblastic tumor mimicking a necrotized malignant mass in a 55-year-old man who was examined with magnetic resonance imaging. To our knowledge, this is the first report of magnetic resonance imaging of mesenteric inflammatory myofibroblastic tumor with extensive central necrosis in an older patient. PMID- 15791486 TI - Radiofrequency ablation of neuroendocrine liver metastases. PMID- 15791487 TI - PCR-based approach for sequencing mitochondrial genomes of decapod crustaceans, with a practical example from kuruma prawn (Marsupenaeus japonicus). AB - An approach for sequencing the entire mitochondrial genomes (mitogenomes) of decapod crustaceans using 79 newly designed and 7 published polymerase chain reaction (PCR) primers is described. The approach comprises the following steps: (1) the entire mitogenome is amplified in 2 or 3 long PCRs; (2) the 86 primers are used in different combinations to amplify contiguous, overlapping short segments of the entire mitogenome with the diluted long PCR products as templates; (3) direct cycle sequencing is conducted using the short PCR products. This strategy allows a more rapid determination of decapod mitogenomic sequences than a traditional method using cloned mitochondrial DNA and primer walking strategy. As a practical example, the mitogenomic sequence for a kuruma prawn Marsupenaeus japonicus (Crustacea: Decapoda), was determined using the PCR-based approach. PMID- 15791488 TI - Isolation and physical mapping of sex-linked AFLP markers in nile tilapia (Oreochromis niloticus L.). AB - Gynogenetically produced XX and YY Nile tilapia (Oreochromis niloticus) and diploid control groups were screened for amplified fragment length polymorphisms (AFLPs) to search for sex-linked or sex-specific markers. Family-level bulked segregant analysis (XX and YY gynogenetic family pools) and individual screening (XX and YY gynogenetics and XX and XY control individuals) identified 3 Y-linked (OniY425, OniY382, OniY227) and one X-linked (OniX420) AFLP markers. OniX420 and OniY425 were shown to be allelic. Single locus polymerase chain reaction assays were developed for these markers. Tight linkage was demonstrated between the AFLP markers and the sex locus within the source families. However, these markers failed to consistently identify sex in unrelated individuals, indicating recombination between the markers and the sex-determining loci. O. niloticus bacterial artificial chromosome clones, containing the AFLP markers, hybridized to the long arm of chromosome 1. This confirmed previous evidence, based on meiotic chromosome pairing and fluorescence in situ hybridization probes obtained through chromosome microdissection, that chromosome pair 1 is the sex chromosomes. PMID- 15791489 TI - First microsatellite loci of red mullet (Mullus barbatus) and their application to genetic structure analysis of Adriatic shared stock. AB - In order to study the genetic structure of the Adriatic shared stock of red mullet (Mullus barbatus), we developed a set of dinucleotide microsatellite markers. A dinucleotide-enriched genomic library was obtained, and 6 polymorphic dinucleotide loci were successfully optimized. The markers showed high expected heterozygosity (from 0.68 to 0.92) and allele number (from 12 to 33); thus they appear to be suitable for detecting genetic differences in the population of red mullet. Four Adriatic samples were subsequently analyzed for microsatellite variation, and the results showed subtle but statistically significant genetic differentiation, indicating that the Adriatic red mullet may group into local, genetically isolated populations. No correlation between geographic distance and genetic differentiation was observed. In addition, the evidence of recent bottlenecks in the Adriatic samples indicates that the observed population subdivision might reflect random local allelic variations, generated by reproductive success, survival rates, or fishing pressure. PMID- 15791491 TI - Population genetic structure of Pacific white shrimp (Litopenaeus vannamei) from Mexico to Panama: microsatellite DNA variation. AB - Genetic variation and population structure of wild white shrimp (Litopenaeus vannamei) from 4 geographic locations from Mexico to Panama were investigated using 5 microsatellite DNA loci. The genetic diversity between populations was indicated by the mean number of alleles per locus and mean observed heterozygosity, which ranged from 7.4 to 8.6 and from 0.241 to 0.388, respectively. Significant departures from Hardy-Weinberg equilibrium were found at most locations at each locus, with the exception Guatemala at Pvan0013, and were caused by high heterozygote deficiencies. Genetic differences between localities were detected by pairwise comparison based on allelic and genotypic frequencies, with the exception of locus Pvan1003. Significant pairwise F (ST) values between locations and total F (ST) showed that the white shrimp population is structured into subpopulations. However, population differentiation does not follow an isolation-by-distance model. Knowledge of the genetic diversity and structure of L.vannamei populations will be of interest for aquaculture and fisheries management to utilize and preserve aquatic biodiversity. PMID- 15791494 TI - [Neuropathic pain: mechanisms, diagnosis and treatment]. AB - Neuropathic pain, resulting from the disturbances of central or peripheral nervous system with mechanisms that we cannot explain yet exactly, is really a troublesome situation both for the patient and the physician. Our knowledge about this difficult to diagnose and treat pain condition has improved a lot with the aid of recent experimental and clinical studies. This review summarizes the underlying mechanisms, common conditions, diagnosis and treatment of neuropathic pain. PMID- 15791490 TI - Isolation and characterization of ornithine decarboxylase gene from flounder (Paralichthys olivaceus). AB - Ornithine decarboxylase (ODC) is a homodimeric enzyme dependent on pyridoxal 5' phosphate. We identified a complementary DNA clone corresponding to ODC from the brain of adult flounder (Paralichthys olivaceus). The flounder ODC cDNA consisted of 2939 bp encoding 272 amino acid residues. The flounder ODC showed 80.3% sequence identity to zebrafish and 70.8% to rat at the amino acid level. Comparison of the structure and nucleotide sequence of the ODC genes revealed that the gene is highly conserved in the flounder, zebrafish, and rat. The presence of ODC mRNA species in brain, kidney, liver, and embryo was confirmed using the reverse transcriptase polymerase chain reaction. The recombinant protein of flounder ODC containing a short histidine tag at the carboxyl terminus was overexpressed in Escherichia coli BL21 (DE3) codon plus using an inducible T7 expression system, and was purified by Ni-NTA affinity chromatography. PMID- 15791492 TI - Molecular analysis of deep-sea hydrothermal vent aerobic methanotrophs by targeting genes of 16S rRNA and particulate methane monooxygenase. AB - Molecular diversity of deep-sea hydrothermal vent aerobic methanotrophs was studied using both 16S ribosomalDNA and pmoA encoding the subunit A of particulate methane monooxygenase (pMOA). Hydrothermal vent plume and chimney samples were collected from back-arc vent at Mid-Okinawa Trough (MOT), Japan, and the Trans-Atlantic Geotraverse (TAG) site along Mid-Atlantic Ridge, respectively. The target genes were amplified by polymerase chain reaction from the bulk DNA using specific primers and cloned. Fifty clones from each clone library were directly sequenced. The 16S rDNA sequences were grouped into 3 operational taxonomic units (OTUs), 2 from MOT and 1 from TAG. Two OTUs (1 MOT and 1 TAG) were located within the branch of type I methanotrophic ?-Proteobacteria. Another MOT OTU formed a unique phylogenetic lineage related to type I methanotrophs. Direct sequencing of 50 clones each from the MOT and TAG samples yielded 17 and 4 operational pmoA units (OPUs), respectively. The phylogenetic tree based on the pMOA amino acid sequences deduced from OPUs formed diverse phylogenetic lineages within the branch of type I methanotrophs, except for the OPU MOT-pmoA-8 related to type X methanotrophs. The deduced pMOA topologies were similar to those of all known pMOA, which may suggest that the pmoA gene is conserved through evolution. Neither the 16S rDNA nor pmoA molecular analysis could detect type II methanotrophs, which suggests the absence of type II methanotrophs in the collected vent samples. PMID- 15791495 TI - Celiac ganglion block. AB - The percutaneus blockade of the celiac plexus is being performed nearly for a century. The aim of performing celiac plexus and splancnic nerve blocks was surgical anesthesia at the beginning. But because of the technical demands and variable results of celiac plexus and splanchnic nerve blocks as a surgical anesthetic, over time, these techniques were supplanted by spinal anesthesia and segmental blockade of the somatic paravertebral nerves. As celiac plexus and splanchnic nerve blocks were falling into disuse for surgical anesthesia, the clinical utility of these techniques was becoming apparent in the new specialty of pain management. Celiac plexus and splancnic nerve blocks are effective in relieving chronic abdominal pain, especially originating from the malignancies of the pancreas, liver, gallbladder, omentum, mesentery, and alimentary tract from the stomach to the transverse portion of the large colon. The relevant anatomy, indications, cotraindications, different application techniques and results of celiac blockade is reviewed in this paper. PMID- 15791496 TI - [Epidural steroid injections and low back pain]. AB - Low-back pain nearly affects all the people in the population; male or female, at any period of their lives and continues with spontaneous regressive attacks. In 10% of the patients, the pain persists despite conservative treatments and causes significant decrements of life quality. For this reason, the patients with chronic low back-pain are applying to pain specialists and treated with different invasive pain therapies. Epidural steroid injections are the most common invasive procedures in the treatment of low-back pain. However, the effects and outcomes are contradictory. Epidural steroid injections should be performed absolutely under fluoroscopic guidance. Pain selection should be made carefully and algorithms should be performed fastidiously. PMID- 15791497 TI - [Challenges in the treatment of pain in children with cancer and tunneling of epidural catheter for long term infusion]. AB - The main differences of cancer pain in children from adults are predominance of tumor related pain, difficulties in pain assessment and rare use of regional blocks. A 2.5 years old girl with severe pain due to pelvic/sacro-cocygeal mass will be presented here. Pain assessment depended on the observations of other patients' parents in the ward, because the child's family was uncooperative. An epidural catheter was placed at T12-L1 level when the conventional methods were unsuccessful and tunneled 5 cm away from the insertion site. Infusion of bupivacain 0.2 mg/kg/hr + fentanyl 0.4 mg/kg/hr from the catheter provided pain relief. Another epidural catheter placed when intravenous morphine infusion (0.02 mg/kg/hr) became inefficient following accidental removal of the initial one. The patient was transferred to another hospital for chemotherapy on the 2nd week. The good interactions between the anesthesiologists accomplished the continuity of pain therapy. Tunneling epidural catheter is an invaluable technique in children who are resistant to conventional pain therapies. PMID- 15791498 TI - [Effect of tramadol on regeneration after experimental sciatic nerve injury]. AB - Abnormal impulses in peripheral nerves play a critical role in neuropathic pain syndromes. The voltage-gated Na+ channels that underlie the action potential are main targets for clinically useful drugs in the pain therapy. Systemic tramadol has been shown to have clinical efficacy against some forms of neuropathic pain. Therefore, we investigated the mechanisms of action of tramadol by an in vitro model by sucrose-gap technique. Tramadol produced concentration-dependent and frequency-dependent decrements in CAP amplitude. Also, injured nerves were more sensitive to tramadol. Tramadol decreased the amplitude of the delayed depolarization and the hyperpolarizing afterpotentials. In conclusion, blocking potencies of small concentration tramadol on the delayed depolarization and hyperpolarizing afterpotential in regeneration period may be contributed for understanding of the action mechanisms of tramadol on neuropathic pain therapy. PMID- 15791499 TI - [The use of transdermal fentanyl in chronic non-malignant pain]. AB - Twelve non-malignant pain patients were followed-up for pain, improvement in the quality of life, satisfaction for pain therapy and side effects for a mean of 95.25 days with the use of transdermal fentanyl (TDF). During this period a mean of 32.04 mg/gr TDF was used. While the score of pain at rest was decreased by 52.6%, mean pain score on movement was decreased by 45.2% (p= 0.002). Quality of sleep improved and impairment of daily living by pain was decreased significantly (p= 0.002). Satisfaction by the pain therapy was 83.3% and the most common side effect was nausea (16.8%). TDF may be a good alternative in the therapy of chronic non-malignant pain if patients were selected carefully. PMID- 15791500 TI - [The relationship of stabbing headaches with migraine attacks]. AB - Temporal and spatial relationships between idiopathic stabbing headache and migraine headache have so far been reported. We aimed to obtain some clues about the stabbing headache pathophysiology by comparing the two types of headache in regard to temporal and spatial relationships as well as precipitating and relieving factors of stabbing headache during migraine attacks. 43 patients who reported temporal relationship between migraine headache and stabs were studied. Localizations of migraine and stabbing headaches overlapped in 38 (88%) of the patients. 34 (79%) had stabs during migraine attacks, while 9 (21%) had stabs with a very close temporal relationship to migraine attacks. During the attacks, while head movements caused stabs in 10 patients (23%); applying pressure to the temples (n=3) and sleeping (n=2) alleviated or abolished stabs. We thought that stabs probably occur during a period in which central pain control mechanisms are weakened. PMID- 15791501 TI - [The effects of clodronate for the pain treatment of bone metastasis due to prostate cancer]. AB - Hormone refractory prostate cancer is dominated by osseous metastases. Bisphosphonates are able to reduce bone resorption. Sixteen hormone refractory prostate cancer patients with related bone metastases were included in the study. Group A consisted of patients who were not treated with bisphosphonates (n=9) and group B consisted of patients who had received bisphosphonates treatment previously, but not receiving currently (n=7). All patients were treated with the same analgesic medications. Clodronate 400 mg; 1200 mg/day (p.o.) was added to the treatment of the patients in group A. Visual Analogue Scale (VAS) scores, consumptions and side effects of analgesics were recorded by two week intervals. Alkaline phosphatase, creatinine and serum Ca++ levels were controlled by 4 week intervals. At the end of the 12th week, the study was ended. In Group A, VAS decreased at the end of the 2nd week but in Group B VAS decreased in the 4th week. VAS decreased 75% in group A and 65.7% in group B and the difference was considered statistically significant (p<0.0001). Clodronate treatment was stopped in 2 patients because of nausea, 7 patients are still being treated with clodronate. We conclude that bisphosphonates treatment of painful osseous metastasis due to hormone refractory prostate cancer results in significant pain decrease. PMID- 15791502 TI - [Low doses of prazosin potantiates the antinociceptive/analgesic effect of amitriptyline]. AB - In this study, we aimed to reveal the interaction between the tricyclic antidepressant amitriptyline and a1-adrenoceptor antagonist prazosin in mice by using an analgesiometric device hot-plate. Amitriptyline (10 mg/kg) has analgesic effect as expected. Neither of the prazosin doses (0.1, 0.2, 0.5, 1 mg/kg) displayed analgesic effect alone. The combination of lower doses of prazosin (0.1, 0.2, 0.5 mg/kg) with amitriptyline (10 mg/kg) potentiated the antinociceptive effect of this drug. However, the relatively higher dose of prazosin (1 mg/kg) did not effect amitriptyline analgesia. Thus we conclude that the antinociceptive effect of amitriptyline is potentiated by low doses of prazosin. PMID- 15791504 TI - Can routine laboratory tests discriminate between severe acute respiratory syndrome and other causes of community-acquired pneumonia? AB - BACKGROUND: The clinical presentation of severe acute respiratory syndrome (SARS) resembles that of other etiologies of community-acquired pneumonia, making diagnosis difficult. Hematological and biochemical abnormalities, particularly lymphopenia, are common in patients with SARS. METHODS: With the use of 2 databases, we compared the ability of the absolute lymphocyte count, absolute neutrophil count, lactate dehydrogenase level, creatine kinase level, alanine aminotransferase level, and serum calcium level at hospital admission to discriminate between cases of SARS and cases of community-acquired pneumonia. The SARS database contained data for 144 patients with SARS from the 2003 Toronto SARS outbreak. The community-acquired pneumonia database contained data for 8044 patients with community-acquired pneumonia from Edmonton, Canada. Patients from the SARS database were matched to patients from the community-acquired pneumonia database according to age, and receiver operating characteristic curves were constructed for each laboratory variable. RESULTS: The areas under the receiver operating characteristic curves (AUCs) demonstrated fair to poor discriminatory ability for all laboratory variables tested except absolute neutrophil count, which had an AUC of 0.80, indicating good discriminatory ability (although there was no cutoff value of the absolute neutrophil count at which reasonable sensitivity or specificity could be obtained). Combinations of any 2 tests did not perform significantly better than did the absolute neutrophil count alone. CONCLUSIONS: Routine laboratory tests, including determination of absolute lymphocyte count, should not be used in the diagnosis of SARS or incorporated into current case definitions of SARS. The role of the absolute neutrophil count in SARS diagnosis is likely limited, but it should be assessed further. PMID- 15791505 TI - Impact of fluoroquinolone prophylaxis on reduced infection-related mortality among patients with neutropenia and hematologic malignancies. AB - BACKGROUND: Fluoroquinolone prophylaxis during neutropenia in patients with cancer has been associated with decreased incidence of gram-negative bacteremia. Bacterial antimicrobial resistance is likely to cause a progressive lack of efficacy of fluoroquinolones, but no convincing evidence from clinicoepidemiologic observations has proved this hypothesis. METHODS: This prospective observational study assessed the impact of discontinuing fluoroquinolone prophylaxis on the incidences of fever and bacteremia and on mortality among patients with neutropenia, after chemotherapy for hematologic malignancies. RESULTS: After a 12-month baseline period of levofloxacin prophylaxis, a period of discontinuation of fluoroquinolone prophylaxis was planned but was stopped prematurely after 9 neutropenic episodes over 3 weeks, because the mortality rate (33.3%) was higher than that with routine fluoroquinolone prophylaxis (2.9%) (odds ratio [OR], 16.6; 95% confidence interval [CI], 3.6-77.2). Fewer patients had gram-negative bacteremia during the baseline period (4.8%; n=15) than during the discontinuation period (44.4%; n=4) (OR, 16.9; 95% CI, 4.1-70.0). After levofloxacin therapy was reintroduced, the incidence of gram-negative bacteremia and the mortality rate were comparable to those during the first period. Escherichia coli isolated during the discontinuation period was susceptible to levofloxacin in vitro, whereas all E. coli isolates isolated during both prophylaxis periods were resistant. Bloodstream infections were caused by a single agent when the patient had received levofloxacin prophylaxis, whereas most cases of gram-negative bacteremia were polymicrobial after discontinuation. CONCLUSIONS: These findings suggest that, despite increasing rates of antimicrobial resistance, levofloxacin prophylaxis during neutropenia may have a beneficial impact on morbidity and infection-related mortality. Continued monitoring of the rate of gram-negative bacteremia is warranted for timely detection of the loss of efficacy of fluoroquinolone prophylaxis. PMID- 15791506 TI - Fluoroquinolone prophylaxis in patients with neutropenia. PMID- 15791507 TI - A prospective, randomized, and comparative study of 3 different methods for the diagnosis of intravascular catheter colonization. AB - BACKGROUND: Demonstration of catheter tip colonization is usually performed by use of Maki's semiquantitative technique, although the superiority of quantitative techniques has been claimed on the basis of their purported ability to detect both endoluminal and exoluminal microorganisms. METHODS: We prospectively compared Maki's semiquantitative technique and the quantitative methods of sonication and vortexing for the detection of colonization of intravascular catheter tips and catheter-related bloodstream infections. All 3 techniques were performed on the tip of each catheter, and the order in which each technique was performed was randomly assigned. RESULTS: Of the 1000 catheter tips that were processed, 329 (32.9%) had positive results for at least 1 of the 3 techniques when a breakpoint of >or=100 colony-forming units (cfu)/catheter segment was used for the quantitative techniques and a breakpoint of >or=15 cfu was used for Maki's technique. Eighty-two of the catheter tips for which results were positive were from patients with catheter-related bloodstream infections. For each technique, the likelihood of detection decreased progressively depending on the order in which the technique was performed (i.e., second vs. first and third vs. second). The likelihood of detection of catheter colonization for each technique, when the technique was performed first and when 2 breakpoints (>or=100 cfu/catheter segment [criterion B] and >or=1000 cfu/catheter segment [criterion A]) were used for the quantitative techniques and a breakpoint of >or=15 cfu was used for Maki's technique, was as follows: 99.1% and 100% for Maki's technique, 95.1% and 92.9% for sonication, and 93.1% and 72.8% for vortexing (for criteria B and A, respectively). No inferiority of Maki's technique could be demonstrated when results were compared according to whether catheter placement was short term (i.e., <7 days) or long term (i.e., >or=7 days), either for the detection of colonization or for the detection of catheter-related bloodstream infections. CONCLUSIONS: According to data from the present study, the quantitative techniques of sonication and vortexing were not superior to Maki's technique under the test conditions used. The greater simplicity of Maki's semiquantitative technique makes it the procedure of choice for routine work in the microbiology laboratory. PMID- 15791508 TI - Clonal groups and the spread of resistance to trimethoprim-sulfamethoxazole in uropathogenic Escherichia coli. AB - BACKGROUND: Antibiotic resistance is increasingly complicating the management of urinary tract infection. We investigated the extent to which a group of Escherichia coli called clonal group A (CGA), which is associated with resistance to trimethoprim-sulfamethoxazole (TMP-SMZ), accounted for TMP-SMZ resistance among a prospectively collected set of uropathogenic and rectal E. coli isolates from a university population in Michigan. METHODS: Resistant and susceptible uropathogenic E. coli isolates (45 each) and 79 randomly selected rectal E. coli isolates were evaluated for CGA status by use of 2 definitions of this group-- the enterobacterial repetitive intergenic consensus sequence 2 (ERIC2)-polymerase chain reaction (PCR) pattern A fingerprint and the C288T single nucleotide polymorphism (SNP) in the fumC gene. We compared virulence gene profiles and molecular mechanisms of resistance to TMP-SMZ between isolates classified as CGA by both approaches to better characterize the relationship between isolates. RESULTS: Of the 45 isolates that exhibited ERIC2-PCR pattern A, one-half (23 of 45) were resistant to TMP-SMZ, and 16 contained the C288T SNP. The pattern A isolates were diverse, exhibiting multiple mechanisms of resistance to TMP-SMZ and various combinations of virulence factors. C288T SNP isolates showed less variation, with 15 of 16 resistant to TMP-SMZ and a 1.8-kb class I integron bearing the dfrA17 gene present in 14 of 15 resistant isolates. Twelve of 16 exhibited the same combination of virulence genes. Pulsed-field gel electrophoresis patterns for these 12 isolates were unique. CONCLUSION: CGA, as defined by the fumC C288T SNP, appears to be distantly clonal but is not an outbreak-related group. The widespread group has likely evolved through lateral transfer of genes conferring virulence and antibiotic resistance. PMID- 15791509 TI - Left ventricular assist device-related infection: treatment and outcome. AB - BACKGROUND: Left ventricular assist device (LVAD) implantation has become an effective treatment option for patients with severe heart failure awaiting transplantation. Significant infection rates have been reported among LVAD recipients. However, few reports have focused specifically on device infection, its treatment, and the impact of LVAD-related infection on clinical outcome. METHODS: Forty-six LVAD-related infections were diagnosed in 38 (50%) of 76 patients who underwent LVAD implantation as a bridge to transplantation. Twenty nine episodes of LVAD-related bloodstream infection (BSI) (including 5 that were cases of LVAD endocarditis) and 17 episodes of local LVAD infection were identified. RESULTS: Diabetes mellitus appeared to increase the risk of BSI among patients with LVAD infection. LVAD-related infection delayed transplantation, as reflected by longer device-support times (a mean duration +/- SEM of 182.8+/-31.1 days, compared with 66.3+/-8.8 days; Por=6 years, during the first 2 years after vaccination (vaccine effectiveness, 95%; 95% CI, 68%-99%; P<.002), and protection remained high during the following 3 years (77%; 95% CI, -364% to 99%; P=.34). For children aged 2-5 years, the estimated effectiveness was positive during the first 2 years (62%; 95% CI, -403% to 97%; P=.47) but was negative during the following period (-74%; 95% CI, -1956% to 85%; P=.66). Among children aged <2 years, there was no evidence of protection. Household crowding and disadvantaged socioeconomic conditions were associated with increased risk of disease. CONCLUSIONS: The polysaccharide vaccine remains a cost-effective option for the short-term protection of school-aged children and adults; however, conjugate vaccines are needed for younger children. PMID- 15791511 TI - Association between Chlamydia pneumoniae and acute myocardial infarction in young men in the United States military: the importance of timing of exposure measurement. AB - BACKGROUND: Several investigators have found that Chlamydia pneumoniae and cytomegalovirus infections may be risk factors for coronary heart disease. However, the data remain controversial. To address this hypothesis, data and specimens were collected from a well-established prospective cohort of active duty personnel from the US military. METHODS: A nested case-control study was conducted with 300 case patients and 300 matched control subjects. Case patients were men (age, 30-50 years) with a medically documented, first-time hospitalization for acute myocardial infarction (MI) and from whom a serum specimen had been drawn >or=1 year before the time of the acute MI. Population based control subjects were chosen from the same cohort and were individually matched by age, race, and time of specimen collection. Evidence of past infections with C. pneumoniae and cytomegalovirus were measured by microimmunofluorescence assay and enzyme-linked immunosorbent assay, respectively. RESULTS: Significant risk was associated with high titer (>or=1 : 64) to C. pneumoniae immunoglobulin A (IgA) (adjusted relative risk [RR(adj)], 1.67; 95% confidence interval [CI], 1.04-2.70). This increased risk was greatest when specimens were collected 1-5 years before the event (RR(adj), 2.11; 95% CI, 1.06-4.21). High titer (>or=1 : 256) to C. pneumoniae immunoglobulin G (IgG) was also associated with an elevated risk (RR(adj), 1.74; 95% CI, 0.90-3.34) after full adjustment for cardiovascular risk factors, whereas no independent risk for acute MI was associated with cytomegalovirus IgG seropositivity. CONCLUSIONS: This study demonstrates a significant association between high titers to C. pneumoniae IgG and IgA and acute MI in a cohort of young men and suggests that recent or chronic active infections could be associated with an increased risk for MI. PMID- 15791512 TI - Chlamydia pneumoniae and atherosclerosis. PMID- 15791513 TI - Safety profile of smallpox vaccine: insights from the laboratory worker smallpox vaccination program. AB - BACKGROUND: The frequency of mild-to-moderate adverse events following smallpox vaccination was not well documented or reported during the pre-eradication era. This report describes the frequency of such symptoms among 936 adult smallpox vaccinees with and without a history of prior smallpox vaccination. METHODS: Diary cards were distributed to 1006 laboratory workers and members of the Centers for Disease Control and Prevention (CDC) smallpox response team who received smallpox vaccination under an investigational new drug protocol during 2001-2002. Vaccinees were requested to complete the diary card daily and return it to the CDC 28 days after vaccination. The proportion of vaccinees reporting symptoms was determined and compared among subgroups. RESULTS: Ninety-three percent of the diary cards were returned. The most common symptom reported was "itching at vaccination site." Primary vaccines reported statistically higher proportions of the following 11 symptoms: joint pain (25% vs. 11%; P=.0011), muscle pain (46% vs. 19%; P<.0001), fatigue (43% vs. 29%; P=.0161), swelling at vaccination site (58% vs. 33%; P<.0001), itching on the body (31% vs. 17%; P=.0048), abdominal pain (11% vs. 2%; P=.0012), swollen or tender lymph nodes (71% vs. 33%; P<.0001), pain at injection site (48% vs. 30%; P=.0018), headache (40% vs. 25%; P=.0088), backache (17% vs. 7%; P=.0090), and fever (temperature, >or=100 degrees F [37.7 degrees C]; 20% vs. 9%; P=.0047). CONCLUSIONS: This analysis suggests that previously unvaccinated persons aged <30 years experienced more symptoms than did previously vaccinated persons. The findings of increased proportions with joint pain, abdominal pain, backache, and difficulty breathing were unexpected. As with recently described cardiac adverse events, these symptoms are suggestive of systemic involvement and warrant further study. PMID- 15791514 TI - Voriconazole for invasive bone aspergillosis: a worldwide experience of 20 cases. AB - BACKGROUND; Bone aspergillosis remains a rare but potentially devastating fungal disease. Although voriconazole is effective for invasive pulmonary aspergillosis, evidence of its efficacy for aspergillosis located in bone is limited. METHODS: We report our experience with voriconazole in 4 cases of invasive bone aspergillosis. In addition, all cases of probable and definite bone aspergillosis from the Pfizer clinical database were reviewed and analyzed to determine the safety and efficacy of voriconazole treatment. Global response was evaluated at the end of therapy on the basis of a composite assessment of overall clinical, radiological, and mycological responses. RESULTS: Twenty patients are described, of whom 18 had definite bone involvement diagnosed (spondylodiskitis in 9, sternum/rib osteomyelitis in 6, and peripheral bone involvement in 5). Of 20 patients, 14 were immunocompromised. Oral or intravenous voriconazole was given as salvage therapy for 18 patients; 2 patients received voriconazole as first line therapy. Median duration of voriconazole treatment was 83.5 days (range, 4 395 days). Global response at end of therapy was satisfactory in 11 (55%) of 20 patients, including complete responses in 4 patients and partial responses in 7 patients; there were no relapses of infection in the 4 patients with complete response to therapy with voriconazole. Treatment was generally well tolerated. CONCLUSIONS: Long-term voriconazole treatment is a new therapeutic option for invasive aspergillosis with bone involvement. PMID- 15791515 TI - Efficacy of thermotherapy to treat cutaneous leishmaniasis caused by Leishmania tropica in Kabul, Afghanistan: a randomized, controlled trial. AB - BACKGROUND: Pentavalent antimony is the agent recommended for treatment of cutaneous leishmaniasis (CL). Its use is problematic, because it is expensive and because of the potential for drug-associated adverse effects during a lengthy and painful treatment course. METHODS: We tested the efficacy of thermotherapy for the treatment of CL due to Leishmania tropica in a randomized, controlled trial in Kabul, Afghanistan. We enrolled 401 patients with a single CL lesion and administered thermotherapy using radio-frequency waves (1 treatment of >or=1 consecutive application at 50 degrees C for 30 s) or sodium stibogluconate (SSG), administered either intralesionally (a total of 5 injections of 2-5 mL every 5-7 days, depending on lesion size) or intramuscularly (20 mg/kg daily for 21 days). RESULTS: Cure, defined as complete reepithelialization at 100 days after treatment initiation, was observed in 75 (69.4%) of 108 patients who received thermotherapy, 70 (75.3%) of 93 patients who received intralesional SSG, and 26 (44.8%) of 58 patients who received intramuscular SSG. The OR for cure with thermotherapy was 2.80 (95% confidence interval [CI], 1.45-5.41), compared with intramuscular SSG treatment (P=.002). No statistically significant difference was observed in the odds of cure in comparison of intralesional SSG and thermotherapy treatments. The OR for cure with intralesional SSG treatment was 3.75 (95% CI, 1.86-7.54), compared with intramuscular SSG treatment (P<.001). The time to cure was significantly shorter in the thermotherapy group (median, 53 days) than in the intralesional SSG or intramuscularly SSG group (median, 75 days and >100 days, respectively; P=.003). CONCLUSIONS: Thermotherapy is an effective, comparatively well-tolerated, and rapid treatment for CL, and it should be considered as an alternative to antimony treatment. PMID- 15791516 TI - Boiling the boil. PMID- 15791517 TI - New considerations in infectious disease outbreaks: the threat of genetically modified microbes. AB - Genetically altered microbes are used widely in the conduct of scientific study and have facilitated the development of new medical therapies, preventive strategies, and diagnostic tools. Unfortunately, such organisms may also cause infectious disease outbreaks as a result of accidental or intentional transmission to humans. The unique microbial properties of genetically altered pathogens and the clinical symptoms exhibited by persons infected with them may impede the usual diagnostic and clinical evaluations or preventive strategies. Practicing physicians and clinicians at microbiology laboratories, who would be the first to observe such infections, must consider the broad clinical possibilities of illnesses caused by deliberately altered microorganisms and the potential difficulty in confirming a diagnosis. PMID- 15791518 TI - Risk of person-to-person transmission of pneumonic plague. AB - Plague has received much attention because it may be used as a weapon by terrorists. Intentionally released aerosols of Yersinia pestis would cause pneumonic plague. In order to prepare for such an event, it is important, particularly for medical personnel and first responders, to form a realistic idea of the risk of person-to-person spread of infection. Historical accounts and contemporary experience show that pneumonic plague is not as contagious as it is commonly believed to be. Persons with plague usually only transmit the infection when the disease is in the endstage, when infected persons cough copious amounts of bloody sputum, and only by means of close contact. Before antibiotics were available for postexposure prophylaxis for contacts, simple protective measures, such as wearing masks and avoiding close contact, were sufficient to interrupt transmission during pneumonic plague outbreaks. In this article, I review the historical literature and anecdotal evidence regarding the risk of transmission, and I discuss possible protective measures. PMID- 15791519 TI - Nitazoxanide: a new thiazolide antiparasitic agent. AB - Nitazoxanide is a new thiazolide antiparasitic agent that shows excellent in vitro activity against a wide variety of protozoa and helminths. It is given by the oral route with good bioavailability and is well tolerated, with primarily mild gastrointestinal side effects. At present, there are no documented drug-drug interactions. Nitazoxanide has been licensed for the treatment of Giardia intestinalis-induced diarrhea in patients >or=1 year of age and Cryptosporidum induced diarrhea in children aged 1-11 years. At present, it is pending licensure for treatment of infection due to Cryptosporidium species in adults and for use in treating immunocompromised hosts. It represents an important addition to the antiparasitic arsenal. PMID- 15791520 TI - Switch from ritonavir to indinavir in combination therapy for HIV-1-infected children. AB - BACKGROUND: Protease inhibitors are an effective component of combination antiretroviral treatment for children infected with human immunodeficiency virus 1 (HIV-1), but tolerance or toxicity issues sometimes require an alternative therapeutic strategy. METHODS: HIV-1-infected children aged 2-17 years received combination therapy with either stavudine plus ritonavir or with zidovudine, lamivudine, and ritonavir as part of a randomized clinical trial. Twenty-one months after the start of the trial, ritonavir in capsule formulation became unavailable. The treatment regimen for 25 children was switched from ritonavir capsules to indinavir capsules (500 mg/m(2) every 8 h). The other study drugs remained unchanged. A matched-pairs analysis was performed to compare the results for these 25 children with the results for 25 matched children whose treatment regimen continued to include ritonavir (in liquid formulation). RESULTS: There were no significant differences in the percentage of children with an HIV-1 RNA load of or =1.54 million years) and transients (with membership times <1.54 million years). During episodes of species loss, there is a preferential disappearance of transients while residents are retained, a pattern referred to as the "seniority rule." The residents define the studied communities. They are associated with early successional stages of vegetation, and transients are associated with later stages. Under stable conditions, early arrivals in succession are "transient" and replaced by competitive later arrivals. The reversed roles of transients and residents in the studied fossil record are explained by assuming high degrees of disturbance. We view the system within the context of nonequilibrium metapopulation theory, in which competitively superior species become transients because of their dependence on ephemeral late successional habitats. PMID- 15791532 TI - Simultaneous Quaternary radiations of three damselfly clades across the Holarctic. AB - If climate change during the Quaternary shaped the macroevolutionary dynamics of a taxon, we expect to see three features in its history: elevated speciation or extinction rates should date to this time, more northerly distributed clades should show greater discontinuities in these rates, and similar signatures of those effects should be evident in the phylogenetic and phylodemographic histories of multiple clades. In accordance with the role of glacial cycles, speciation rates increased in the Holarctic Enallagma damselflies during the Quaternary, with a 4.25x greater increase in a more northerly distributed clade as compared with a more southern clade. Finer-scale phylogenetic analyses of three radiating clades within the northern clade show similar, complex recent histories over the past 250,000 years to produce 17 Nearctic and four Palearctic extant species. All three are marked by nearly synchronous deep splits that date to approximately 250,000 years ago, resulting in speciation in two. This was soon followed by significant demographic expansions in at least two of the three clades. In two, these expansions seem to have preceded the radiations that have given rise to most of the current biodiversity. Each also produced species at the periphery of the clade's range. In spite of clear genetic support for reproductive isolation among almost all species, mtDNA signals of past asymmetric hybridization between species in different clades also suggest a role for the evolution of mate choice in generating reproductive isolation as species recolonized the landscape following deglaciation. These analyses suggest that recent climate fluctuations resulted in radiations driven by similar combinations of speciation processes acting in different lineages. PMID- 15791533 TI - Amino acids in nectar enhance butterfly fecundity: a long-awaited link. AB - Thirty years ago, researchers discovered that flowers pollinated by butterflies are consistently rich in nectar amino acids, and more recent findings have shown that butterflies prefer nectar with high amino acid content. These observations led to speculation that amino acids in nectar enhance butterfly fitness and that butterflies have acted as agents of natural selection on nectar composition. Despite a number of experimental efforts over the years, convincing proof that nectar amino acids affect butterfly fitness has been lacking. Here, we provide the first evidence that amino acids in nectar have a positive effect on fecundity of one butterfly species, supporting the existence of a relationship between nectar preferences and fitness benefits. Map butterflies (Araschnia levana L.) raised under natural larval food conditions laid more eggs when they were fed nectar containing amino acids, whereas nectar amino acids had no effect on the number of eggs laid by butterflies raised on larval food rich in nitrogen. Uptake and utilization of nectar amino acids by map butterflies appear to be compensatory mechanisms enabling them to override impacts of poor larval food. These results provide strong support for the long-standing postulate that nectar amino acids benefit butterflies. PMID- 15791534 TI - Evolution of sexual isolation during secondary contact: genotypic versus phenotypic changes in laboratory populations. AB - We monitored the phenotypic and genotypic changes that occur when two behavioral races come into contact in laboratory populations. Drosophila melanogaster from Zimbabwe and nearby regions (Z type) show strong but asymmetric sexual isolation from their cosmopolitan counterparts (M type). Crosses of Z females and M males do not take place readily when other choices are available. At least 15 loci are known to control Z-type mating preferences and performance. By thoroughly mixing the genomes of the two types in laboratory populations, we artificially created maximum secondary contact. Despite the strength of sexual selection favoring Z type male characters, Z-type behavior is eliminated or greatly diminished in all 12 hybrid populations after only 60 generations. This trend is consistent with the spread of the M-type behavior throughout the world as well as a detailed analysis of fitness components. Surprisingly, in contrast with the phenotypic convergence toward the M-type, genotypic samples broadly covering the genomic regions of mapped behavioral loci show no such trend. The genome appears to be "fine grained," with adjacent loci having different evolutionary dynamics and genealogical histories. PMID- 15791535 TI - Evolution of sexual isolation in laboratory populations: fitness differences between mating types and the associated hybrid incompatibilities. AB - Drosophila melanogaster is known to have two races in the incipient stages of speciation that exhibit strong asymmetric premating isolation: Zimbabwe (Z) and cosmopolitan (M). In a study examining the phenotypic and genotypic evolution after secondary contact, we found that despite strong sexual selection favoring the Z-type behavior, it is the M-type behavior that comes to dominate hybrid populations. This article examines the fitness costs associated with the Z-type behavior. We have discovered that these costs are great enough to explain the failure of the Z-type behavior to prosper. Here we report that Z-type females produce approximately half the number of offspring that M-type females produce. Furthermore, crosses between populations have revealed that Z-type females mated to M-type males have approximately 20% fewer offspring than the reciprocal crosses because of an inability of M-type sperm to successfully fertilize Z-type eggs. Hybrid crosses also exhibit much-reduced numbers of viable offspring in addition to reduced hybrid male fertility. These fitness effects suggest that multiple mechanisms of postmating isolation have evolved concurrently with the divergence in behavior. PMID- 15791536 TI - A critical review of twenty years' use of the resource-ratio theory. AB - A model of species interactions based on their use of shared resources was proposed in 1972 by Robert MacArthur and later expanded in an article (1980) and a book (1982) by David Tilman. This "resource-ratio theory" has been used to make a number of testable predictions about competition and community patterns. We reviewed 1,333 papers that cite Tilman's two publications to determine whether predictions of the resource-ratio theory have been adequately tested and to summarize their general conclusions. Most of the citations do not directly test the theory: only 26 studies provide well-designed tests of one or more predictions, resulting in 42 individual tests of predictions. Most of these tests were conducted in the laboratory or experimental microcosms and used primary producers in freshwater systems. Overall, the predictions of the resource-ratio theory were supported 75% of the time. One of the primary predictions of the model, that species dominance varies with the ratio of resource availabilities, was supported by 13 of 16 tests, but most other predictions have been insufficiently tested. We suggest that more experimental work in a variety of natural systems is seriously needed, especially studies designed to test predictions related to resource supply and consumption rates. PMID- 15791537 TI - The impact of directed versus random movement on population dynamics and biodiversity patterns. AB - An improved understanding of dispersal behavior is needed to predict how populations and communities respond to habitat fragmentation. Most spatial dynamic theory concentrates on random dispersal, in which movement rates depend neither on the state of an individual nor its environment and movement directions are unbiased. We examine the neglected dispersal component of directed movement in which dispersal is a conditional and directional response of individuals to varying environmental conditions. Specifically, we assume that individuals bias their movements along local gradients in fitness. Random movers, unable to track heterogeneous environmental conditions, face source-sink dynamics, which can result in deterministic extinction or increase their vulnerability to stochastic extinction. Directed movers track environmental conditions closely. In fluctuating environments, random movers "spread their bets" across patches, while directed movers invest offspring in habitats currently enjoying propitious conditions. The autocorrelation in the environment determines each strategy's success. Random movers permeate entire landscapes, but directed movers are more geographically constrained. Local information constraints limit the ranges of directed movers and introduce a role for historical contingency in determining their ultimate distribution. These geographic differences have implications for biodiversity. Random movement maintains biodiversity through local coexistence, but directed movement favors a spatial partitioning of species. PMID- 15791538 TI - Host specialization and geographic localization of avian malaria parasites: a regional analysis in the Lesser Antilles. AB - We recovered 26 genetically distinct avian malaria parasite lineages, based on cytochrome b sequences, from a broad survey of terrestrial avifauna of the Lesser Antilles. Here we describe their distributions across host species within a regional biogeographic context. Most parasite lineages were recovered from a few closely related host species. Specialization on one host species and distribution across many hosts were both rare. Geographic patterns of parasite lineages indicated limited dispersal and frequent local extinction. The central islands of the archipelago share similar parasite lineages and patterns of infection. However, the peripheral islands harbor well-differentiated parasite communities, indicating long periods of isolation. Nonetheless, 20 of 26 parasite lineages were recovered from at least one of three other geographic regions, the Greater Antilles, North America, and South America, suggesting rapid dispersal relative to rate of differentiation. Six parasite lineages were restricted to the Lesser Antilles, primarily to endemic host species. Host differences between populations of the same parasite lineage suggest that host preference may evolve more rapidly than mitochondrial gene sequences. Taken together, distributions of avian malarial parasites reveal evidence of coevolution, host switching, extinction, and periodic recolonization events resulting in ecologically dynamic as well as evolutionarily stable patterns of infection. PMID- 15791539 TI - Constraint and competition in assemblages: a cross-continental and modeling approach for ants. AB - The mechanisms leading to structure in local assemblages are controversial. On the one hand, assemblage structure is thought to be the outcome of local interactions determined by the properties of species and their responses to the local environment. Alternatively, this structure has been shown to be an emergent property of assemblages of identical individuals or of random sampling of a regional assemblage. In ants at baits, a combination of environmental stress and interspecific competition is widely held to lead to a unimodal relationship between the abundance of dominant ants and species richness. It is thought that in comparatively adverse environments, both abundance and richness are low. As habitats become more favorable, abundance increases until the abundance of dominant ants is so high that they exclude those that are subordinate and so depress richness. Here we demonstrate empirically that this relationship is remarkably similar across three continents. Using a null model approach, we then show that the ascending part of the relationship is largely constrained to take this form not simply as a consequence of stress but also as a result of the shape of abundance frequency distributions. While the form of the species-abundance frequency distribution can also produce the descending part of the relationship, interspecific competition might lead to it too. Scatter about the relationship, which is generally not discussed in the literature, may well be a consequence of resource availability and environmental patchiness. Our results draw attention to the significance of regional processes in structuring ant assemblages. PMID- 15791540 TI - Common garden and natural selection experiments support ecotypic differentiation in the Dominican anole (Anolis oculatus). AB - The theory behind ecotypic differentiation and ecological speciation assumes a predominant role for natural selection working on characteristics with genetic variance, but experimental support for these assumptions is limited. Lesser Antillean anoles show marked ecotypic variation within islands and the potential for ecological speciation. Common garden rearing experiments on the Dominican anole (Anolis oculatus) suggest that the characters showing geographic variation have genetic variance and are not primarily determined by environmental plasticity. Replicated natural selection experiments using large-scale enclosures show that translocated montane samples experience significant (multivariate) directional selection in both wet and dry seasons in both males and females. The targets of selection appear to be spread among the various character systems. An experiment on 12 geographically segregated populations along a coastal xeric montane rainforest gradient (four replicate enclosures) clearly showed that the magnitude of the directional selection intensity is positively related to the position along this gradient. The results of the common garden and natural selection experiments support the interpretation that the geographic differentiation is primarily driven by natural selection and are compatible with the potential for ecological speciation in this system. PMID- 15791541 TI - Insular carnivore biogeography: island area and mammalian optimal body size. AB - Patterns of size variation in insular mammals have been used to support the claim that mammals have a single optimal body size. This hypothesis enjoys wide support, despite having been questioned on both theoretical and empirical grounds. It is claimed that species of optimal size maintain the highest population densities. Therefore these species are thought to inhabit the smallest islands, where larger and smaller species are generally absent. We sought such a pattern by testing how area affects the body sizes of the largest and smallest carnivore species on islands. Using data on carnivores from 322 islands, we found that the sizes of carnivores on small islands tend to be close to the order's mode. Furthermore, we found that the size distribution of carnivore species that inhabit islands resembles that of those whose range is entirely continental. We conclude that insular carnivores provide no support for theories proposing a single optimal size, and we suspect such theories are also flawed on theoretical grounds. PMID- 15791542 TI - Increased level of linkage disequilibrium in rural compared with urban communities: a factor to consider in association-study design. AB - Few studies have investigated genetic differentiation within nonisolate European populations, despite the initiation of large national sample collections such as U.K. Biobank. Here, we used short tandem repeat markers to explore fine-scale genetic structure and to examine the extent of linkage disequilibrium (LD) within national subpopulations. We studied 955 unrelated individuals of local ancestry from nine Scottish rural regions and the urban center of Edinburgh, as well as 96 unrelated individuals from the general U.K. population. Despite little overall differentiation on the basis of allele frequencies, there were clear differences among subpopulations in the extent of pairwise LD, measured between a subset of X linked markers, that reflected presumed differences in the depths of the underlying genealogies within these subpopulations. Therefore, there are strategic advantages in studying rural subpopulations, in terms of increased power and reduced cost, that are lost by sampling across regions or within urban populations. Similar rural-urban contrasts are likely to exist in many other populations with stable rural subpopulations, which could influence the design of genetic association studies and national biobank data collections. PMID- 15791545 TI - Causes of telengiectasias, reticular veins, and varicose veins. AB - Study of surgical specimens and direct observation by angioscopy has revealed that the varicose venous wall, the valvular annulus, and the valves themselves undergo profound changes. Morphologic investigations have shown dilation of the valve annulus, bulging valve leaflets, commissural dilation, leaflet stretching, and eventually complete destruction of the valves. The venous wall has been seen to undergo changes of thickening in some segments and thinning in others. Our investigations show that inflammation and subsequent remodeling of the venous valves and wall are the fundamental mechanisms underlying the observed lesions. Hemodynamic forces, such as blood pressure changes in the wall and sheer stress, as well as varying planes of laminar and turbulent flow, induce activation of leukocytes and endothelial cells. Integrins appear to act as intermediaries and expression of adhesion molecules has been observed. Breakdown of extracellular matrix of the media and adventitia through activation of matrix metalloproteases (MMP) has been observed. In particular, expressions of MMP-1, MMP-2, MMP-9, and tissue inhibitor of metalloproteinase have been studied. Telangiectasias, reticular veins, and true varicose veins appear to be a consequence of the changes induced by venous hypertension and sheer stress. PMID- 15791546 TI - Duplex evaluation of venous insufficiency. AB - Duplex ultrasound is the most useful examination for the evaluation of venous valvular incompetence. Multi-frequency 4 to 7-MHz linear array transducers are typically used for this assessment of superficial and deep reflux. The examination is done with the patient standing and manual compression maneuvers are used to initiate reflux. Automatic rapid inflation and deflation cuffs may be used when a standard stimulus is needed. Cutoff values for reflux have been defined. Perforating veins must be identified and flow direction during compression recorded. When ulcers are present, duplex ultrasound is used to investigate veins of the ulcerated legs. Venous outflow obstruction is also studied by duplex ultrasound and chronic changes in deep and superficial veins following deep venous thrombosis noted. The main drawback in evaluation of chronic obstruction is inability to quantify hemodynamic significance. Anatomic variations in superficial and deep veins are common and their identification is necessary. Reporting results of duplex ultrasound studies must take into consideration the proper classification of venous disease as well as the new anatomic terms that have been accepted. PMID- 15791543 TI - Saami and Berbers--an unexpected mitochondrial DNA link. AB - The sequencing of entire human mitochondrial DNAs belonging to haplogroup U reveals that this clade arose shortly after the "out of Africa" exit and rapidly radiated into numerous regionally distinct subclades. Intriguingly, the Saami of Scandinavia and the Berbers of North Africa were found to share an extremely young branch, aged merely approximately 9,000 years. This unexpected finding not only confirms that the Franco-Cantabrian refuge area of southwestern Europe was the source of late-glacial expansions of hunter-gatherers that repopulated northern Europe after the Last Glacial Maximum but also reveals a direct maternal link between those European hunter-gatherer populations and the Berbers. PMID- 15791547 TI - The long saphenous vein: to strip or not to strip? AB - As 80% of varicose veins stem from incompetence of the great saphenous system, surgeons must choose between simple high ligation of the saphenofemoral junction with avulsion of varicosities or the same procedure done with additional stripping of the long saphenous vein. Many surgeons strip the long saphenous vein by using variations of Keller's 1905 report of inversion stripping. Others may use other techniques. When surgical procedures for varicose veins are compared, they are often measured by the recurrence rate. This, of course, is difficult to define. Comparisons of saphenous vein stripping versus ligation favor the results achieved by stripping but patient satisfaction appears to be equal and while stripping appears to give better satisfaction with regard to quality of life scores, in fact, the risk of nerve damage and subsequent litigation is quite real when the operation of stripping is done. PMID- 15791548 TI - Saphenous ablation: what are the choices, laser or RF energy. AB - Endovenous ablation has been reported to be safe and effective in eliminating the proximal portion of the great saphenous vein from the venous circulation, with faster recovery and better cosmetic results than surgical stripping. However, the definition of a successful outcome in the literature has not been uniform. As in a successful stripping procedure, complete elimination of at least the proximal portion of the great saphenous vein should also be the standard for these endovenous ablation procedures. Our experience with over 1,400 endovenous ablation procedures, of which 1,150 were radiofrequency and over 250 were laser procedures, has allowed evaluation and comparison of these two techniques. And while we have not seen as high success rates as in published reports (especially with laser ablation), we have still concluded that both radiofrequency and laser techniques to destroy the saphenous vein are safe and effective. Patient acceptance is overwhelmingly better than stripping. Physicians performing these techniques should embrace a commitment to addressing all sites of venous insufficiency in a patient, not just the proximal great saphenous vein. Without this level of commitment, one will be left with poor results and a dissatisfied patient. PMID- 15791549 TI - Saphenous ablation: sclerosant or sclerofoam? AB - During the last half of the 20th century, sclerotherapy as a major treatment of varicose veins came and went. At first, it was widely heralded as a substitute for surgery but after a prospective randomized study by Hobbs, interest in sclerotherapy waned. Just before the turn of the 21st century, Cabrera published his experience with foamed sclerosant in patients with great saphenous varices and arteriovenous malformations. Cabrera designed his treatment with the specific aim of obliterating the saphenous trunks. His technique consisted of filling the great saphenous vein in the thigh or the small saphenous vein in the calf with foamed sclerosant injected under ultrasound control. His initial report on long term follow-up revealed that the results were at least comparable to surgery and perhaps somewhat better and his results have been confirmed by others. Investigations into treatment of small vein varices, including telangiectasias, has resulted in the finding that foam results in a 20% improved appearance compared to liquid sclerosant. The most popular sclerosants currently used as foams are polidocanol and sodium tetradecyl sulfate and of the many techniques used in making foams, the technique of Tessari has proven most popular. No randomized clinical trial comparing this technique to surgery has been published; however, the clinical series reports indicate that 80% to 90% of saphenous trunks remain occluded after 3 years when treated by foam sclerotherapy. Complications are seldom encountered but significant skin darkening and superficial thrombophlebitis are common. Temporary vision changes have occurred after foam and liquid sclerotherapy, with foam, these are always transient. Although the long-term efficacy of foamed sclerotherapy treatment is unlikely to be established for years, a number of phlebologists have taken up the practice because it has the advantage of not requiring general or regional anesthesia and takes much less time than equivalent surgical techniques. PMID- 15791550 TI - Foam sclerotherapy: an overview of use for primary venous insufficiency. AB - Ultrasound-guided sclerotherapy with foam is able to suppress blood flow in almost all types of varicose veins. The technique is efficient, simple, safe, inexpensive, and easily repeatable, although it requires training and skill. It should be considered as the best primary treatment for most varicose patients. Evaluation of long-term results, with regard to comparison with other techniques, should take into account all advantages and drawbacks and evaluate cost effectiveness from a life-long point of view. PMID- 15791551 TI - Causes of severe chronic venous insufficiency. AB - A large number of adults in this country have some form of chronic venous insufficiency and a significant percentage of these have venous ulcers. The past decade has refined understanding of leukocyte-mediated injury and has elucidated the role of inflammatory processes in the dermal pathology of chronic venous insufficiency. Understanding of these pathologic cellular functions and molecular regulation of these processes is increasing. PMID- 15791552 TI - Compression therapy for chronic venous insufficiency. AB - Compression therapy provides a means to treat venous stasis, venous hypertension, and venous edema. Different methods of compression therapy have been described periodically over the last 2,000 years. In addition to static compression, specialized compression pumps have been developed and a technique of massage called manual lymphatic drainage has emerged to treat primary and secondary lymphedema. Objectives of compression therapy are to reduce the swollen limb to minimum size, maintain that size, and allow the patient to participate in the care of his limb whenever possible. Reduction therapy is achieved by limb elevation, compression pumps as necessary, and compression wraps. Maintenance therapy largely consists of compression wraps or compression stockings. Nonelastic devices have found a place in treating severe lymphedema but it should be emphasized that periodic follow-up must be done during maintenance therapy so that adjunctive maintenance measures can be added as needed. PMID- 15791553 TI - Superficial vein surgery and SEPS for chronic venous insufficiency. AB - Venous insufficiency in its severe forms leads to skin changes which, in turn may be treated by surgical therapy. Interventions are directed towards correction of the underlying abnormal venous physiology. This involves removal of varicose veins and ablation of incompetent axial veins and relevant perforating veins. In performing ablation of saphenous vein reflux, techniques include high ligation with stripping, radiofrequency ablation, endovenous laser therapy, and foam sclerotherapy. Incompetent perforator interruption can be accomplished surgically by subfascial endoscopic perforator surgery (SEPS) or controlled sclerotherapy using ultrasound. A variety of techniques have emerged to manage the varicose veins themselves. Surgical treatment of chronic venous insufficiency with high ligation in the groin and inversion stripping of the great saphenous vein to the knee combined with stab avulsion of varicose veins continues to be the standard in treatment of varicose veins. There are few comparisons of sclerotherapy of perforating veins with SEPS, but SEPS has become the most popular of surgical options. PMID- 15791554 TI - Severe chronic venous insufficiency: primary treatment with sclerofoam. AB - Venous insufficiency, for practical purposes, can be divided into primary venous insufficiency and chronic venous insufficiency. The latter is characterized by advanced skin changes of hyperpigmentation, edema, ulceration, scarring from healed ulcers or open ulcerations. These are summarized in the CEAP classification as Classes 4, 5 and 6. Pretreatment evaluation is done with a standing ultrasound reflux examination. Thorough mapping of the extremity reflux is desirable. Physiologic tests of venous function, such as plethysmography, are unnecessary. Treatment is directed at closing refluxing axial veins as well as controlling those perforating veins with outward flow. Varicose veins contribute to axial reflux and must be obliterated. Arterial occlusive disease may complicate venous ulceration in as many as 15% of cases. Initial treatment of severe chronic venous insufficiency is usually carried out by controlling the edema with elastic bandaging or nonelastic support, such as the Unna boot or the CircAid dressing. Surgical intervention has been successful but the advent of foam sclerotherapy has proven to be an attractive alternative to surgery and has added a new tool for the treatment of severe chronic venous insufficiency. In this preliminary experience, the results are quite satisfactory and the technique has been shown to be effective, pain-free, inexpensive, with very little morbidity. Guidelines for obtaining sclerosants for use in foam sclerotherapy legally are provided. PMID- 15791556 TI - Introduction: Adequacy of dialysis: problems and challenges. PMID- 15791558 TI - Dialysis dose as a determinant of adequacy. AB - The intent-to-treat analyses of all patients in the HEMO trial suggested that increases in dose of dialysis as measured by urea Kt/V were of marginal or no benefit when dialysis was provided in a 3 times/wk schedule. The as-treated analysis in the HEMO trial pointed to markedly increased mortality when the delivered dose decreased even slightly below the targeted dose, evidence of a dose-targeting bias. The intent-to-treat HEMO study results suggested a potential interaction between sex and the dose-mortality relationship, and this also has been found in some cross-sectional studies, the cause of which remains unexplained. Whether dialysis dose should continue to be targeted based on urea distribution volume (V), or targeted to a body size measure that is a lower power of body weight (such as body surface area), remains an open question. The lack of benefit of increasing the dialysis dose in a 3 times/wk setting is more understandable if one looks at measures of equivalent continuous solute removal, such as the standard Kt/V. Differences in standard Kt/V in the 2 dose arms of the HEMO trial, for example, were only about 15%. Without going into removal of very large solutes (eg, beta-2-microglobulin), which is discussed elsewhere in this issue, or protein-bound uremic solutes, the only way to provide significantly more dialysis dose may be to move to more frequent dialysis schedules and/or to very long session lengths. Here, benefit may be related as much to better control of salt and water balance as to better removal of uremic toxins. PMID- 15791557 TI - Dialysis outcomes as a measure of adequacy of dialysis. AB - Truly adequate dialysis would restore patients to full health, with functional status and length of life indistinguishable from others of the same age, sex, and race without chronic kidney disease. We are far from achieving such outcomes, however, in part because of the dearth of available evidence on which areas of care should be emphasized to get the greatest clinical and psychosocial benefits at the most affordable costs. A clear understanding of the strengths and limitations of currently available evidence can help guide researchers and clinicians in this field, and likely will lead to increasing emphasis on identification and management of comorbid conditions and a focus on preventative medicine. Optimal dialysis will be accomplished only when normal kidney functions are mimicked by artificial devices to a much greater extent than is currently the case. PMID- 15791559 TI - Dialyzer membranes as determinants of the adequacy of dialysis. AB - Hemodialysis membranes have undergone a gradual but substantial evolution over the past few decades. Classification of modern dialyzer membranes by chemical composition bears little relationship to their functional characteristics. The fundamental properties that determine the capacity of the membrane to remove solutes and fluids are its surface area, thickness, pore size, pore density, and potential to adsorb proteins. Dialyzer membrane performance is characterized clinically by its efficiency, defined as the potential to remove urea and presented as the mass-transfer area coefficient (KoA) and ultrafiltration coefficient (K(uf) ),defined as the potential to remove water adjusted for the transmembrane pressure. The parameter K(uf) usually, but not invariably, correlates with the membrane permeability, defined as the potential to remove middle molecules, with beta2-microglobulin being the currently popular marker. The sieving coefficient reflects the membrane potential to transport solutes by convection and is particularly useful for hemofiltration. Enhancing solute clearance is accomplished clinically by increasing blood and dialysate flow rates, strategies that also are applicable to middle molecules for highly permeable membranes. Novel designs of dialyzers include the optimization of fluid flow path geometry and increasing the membrane pore selectivity for solutes by using nanotechnology. PMID- 15791560 TI - Dialysis session length ("t") as a determinant of the adequacy of dialysis. AB - Several studies have shown an association between the hemodialysis session length (the t of Kt or Kt/V) and favorable outcomes for patients on maintenance hemodialysis. In a single randomized controlled trial that systematically varied hemodialysis session length, shorter session length was associated with an increased risk for morbidity and mortality, independent of the time-averaged concentration of urea. Observational studies of dialysis session length have yielded conflicting results, although virtually all studies have confounded hemodialysis session length with hemodialysis efficiency or dose. Limited observational data from nocturnal hemodialysis programs more strongly suggest an independent beneficial effect of longer session length. In aggregate, data on the effects of hemodialysis session length are inconclusive. Future studies should evaluate hemodialysis session length independent of efficiency, and should consider the evaluation of dose by using other clearance parameters and the adequacy of ultrafiltration in addition to solute kinetics. PMID- 15791561 TI - Vascular access as a determinant of adequacy of dialysis. AB - Vascular accesses consist of permanent arteriovenous (AV) accesses (autogenous fistulas and synthetic grafts) and venous accesses (central venous catheters [CVCs]). AV accesses have fewer complications than venous accesses, and are therefore the preferred hemodialysis access. An important additional issue is whether the type of access influences adequacy of dialysis (i.e. Kt/V). Key limiting factors in delivering adequate Kt/V are blood pump speed (Q(B) ), access recirculation, and treatment time. In general, AV accesses support higher Q(B)S with less negative inflow arterial pressures than CVCs. Well-functioning AV accesses are also less likely to exhibit recirculation. Nevertheless, recirculation commonly develops when AV accesses (usually grafts) develop stenosis with decreased access blood flow. Although extension of treatment time can offset the effects of reduced Q(B) and recirculation, this is often impractical and poorly accepted by patients. In conclusion, AV accesses are superior to venous accesses because they are less prone to complications and are more likely to deliver prescribed Kt/V within prescribed treatment time. PMID- 15791562 TI - Dialysis water as a determinant of the adequacy of dialysis. AB - Hemodialysis patients are exposed to large volumes of water in the form of dialysate. Contaminants from the dialysate may cross the dialyzer membrane into the blood and have the potential to compromise the adequacy of dialysis. Several chemicals found commonly in drinking water have long been known to be toxic to hemodialysis patients. More recently, it has become apparent that even low levels of bacterial products in dialysate may adversely impact dialysis adequacy through their ability to stimulate an inflammatory response. Minimum levels of water and dialysate quality have been recommended to protect patients from chemical and microbiologic contaminants. Complying with these recommendations requires an appropriately designed water purification and distribution system, combined with a surveillance program designed to maintain dialysate quality. PMID- 15791563 TI - Adequacy of dialysis: the patient's role and patient concerns. AB - The patient's role in adequacy of hemodialysis is demanding and complex. It requires meticulous attention to initiating, accepting, and maintaining extraordinary behavioral change. This includes the following: (1) major alteration of dietary habits, often contrary to a patient's familial and cultural customs; (2) compliance with a new, voluminous medication routine, often straining personal finances; (3) reallocation of time for transportation, treatment, and partial recovery, frequently consuming a minimum of 6 to 8 hours 3 days each week; (4) psychologic adjustment to unaccustomed chronic dependency on, and accountability to, an array of variably experienced and competent renal care staff; (5) skills, seldom taught, required to communicate clearly and regularly with overworked medical professionals who are often much younger with less life experience; and (6) additional commitment to compensating for the physical fatigue that routinely accompanies hemodialysis. Reasonable behavioral modification in these 6 categories is likely to increase the chances of a patient fulfilling his role in adequacy of dialysis. Some patients, however committed the staff have been in assisting them, may show little interest in dialysis adequacy and the patient's role. Other patients periodically may fail in their role unless the renal care team recognizes the patient as an individual who is included as an important team member. The patient requires consistent and repeated education about their disease, treatment, and risks and benefits of adherence. The unique, unnatural requirements of adequate chronic hemodialysis require this patient support from the renal staff, enhanced by continuous sensitive attention, empathy, and persuasion. This will help the patient achieve success in their role. PMID- 15791564 TI - Adequacy of dialysis in acute renal failure. AB - Renal replacement therapy (RRT) is currently the mainstay of management for patients with acute renal failure (ARF). Adequacy of dialysis in the setting of renal failure is defined poorly and encompasses multiple domains of clinical and biochemical outcomes. Multiple operational factors influence the delivery of adequate dialysis. No current standards exist for RRT for ARF; current RRT practices for ARF generally have been extrapolated from end-stage renal disease (ESRD) literature. The heterogeneity of patient population, variation in RRT practices, and differences in outcomes studied have made it difficult to define or study adequate dialysis in ARF or its impact on clinical outcomes. PMID- 15791565 TI - beta-catenin and its regulatory network. PMID- 15791566 TI - Staging and grading of chronic gastritis. AB - Chronic gastritis is an inflammatory condition of the gastric mucosa that may include structural alterations of the glandular compartment. The semiquantitative scoring systems advocated in the Sydney Systems and the subsequent Atrophy Club Guidelines remain essential for the recognition of the spectrum of the lesions detectable in gastric inflammatory disease. Most practicing pathologists, however, find them too cumbersome to use in their routine diagnostic activities. In this article, we propose a reporting system for chronic gastritis in staging and grading. Staging would convey information on the topography and extension of the gastric atrophic changes, whereas grading should represent the semiquantitative assessment of the combined severity of both mononuclear and granulocytic inflammation. This system could offer gastroenterologists a more immediate perception of the overall condition of the gastric mucosa while also providing useful information about gastric cancer risk. PMID- 15791567 TI - Abnormal beta-catenin expression in oral cancer with no gene mutation: correlation with expression of cyclin D1 and epidermal growth factor receptor, Ki 67 labeling index, and clinicopathological features. AB - Beta-Catenin not only acts as a regulator of E-cadherin-mediated cell-cell adhesion but also plays an important role in Wnt signaling. To assess the prevalence of Wnt signaling, we examined beta-catenin mutation and its immunohistochemical protein expression in oral cancers. The results were linked with expression of cyclin D1, one of the target genes of Wnt signaling, expression of epidermal growth factor receptor (EGFR) relevant to beta-catenin tyrosine phosphorylation, Ki-67 labeling index, clinicopathological features, and survival. In the analysis based on membranous expression of beta-catenin, 75 (68.2%) of 110 cases showed a reduced membranous pattern, and the remaining 35 (31.8%) had a preserved membranous pattern similar to that in oral epithelium. In the analysis of another category of beta-catenin expression, a cytoplasmic/nuclear pattern was observed in 21 (19.1%) of the 110 tumors. Most (19/21, 90.5%) of these tumors had a concomitant reduction of membranous expression of beta-catenin. The reduced membranous or cytoplasmic/nuclear pattern of beta-catenin was significantly associated with an invasive growth pattern, EGFR expression, an increased Ki-67 labeling index, and shorter survival but not with cyclin D1 expression. Mutational analyses of beta-catenin were performed for 39 cases, including the 21 tumors with a cytoplasmic/nuclear pattern, but no mutations in the beta-catenin gene exon 3 were detected in these samples. Our data indicate that altered expression of beta-catenin may play an important role in tumor progression through increased proliferation and invasiveness under EGFR activation. However, mutations of beta-catenin do not appear to be responsible for tumor development and abnormal expression of beta-catenin in oral cancers. PMID- 15791568 TI - Expression and mutational analysis of tyrosine kinase receptors c-kit, PDGFRalpha, and PDGFRbeta in ovarian cancers. AB - Most women with epithelial ovarian cancer are diagnosed with advanced disease. Despite surgery and initial tumor reduction by standard chemotherapy, the tumors frequently recur and the patients eventually die of their disease. New drugs that inhibit tyrosine kinase receptors (TKRs) are being investigated for treatment and this study was undertaken to determine the expression and mutational state for 3 TKRs (c-kit, platelet-derived growth factor receptor [PDGFR] alpha, and PDGFR beta) in ovarian cancer. Tissue arrays containing 84 epithelial ovarian tumors were studied by immunohistochemistry with antibodies specific for c-kit, PDGFR alpha, and PDGFR beta. Immunoreactivity was detected in 78% of the tumor to at least one TKR. PDGFR alpha was expressed in the largest percentage of ovarian tumors (58%) whereas 29% expressed PDGFR beta. Two commercial antibodies against c-kit were studied and 33% of the tumors stained with one but only 6% were interpreted as positive with the second antibody. Activation of TKRs may occur through mutations but, by sequence analysis, no mutations were detected in 6 ovarian tumors with elevated immunoreactivity for each of the TKRs (c-kit, PDGFR alpha, and PDGFR beta). Tyrosine kinase receptors could also be activated through autocrine or paracrine stimulation of receptor by its ligand. Of 43 (35%) tumors tested for both c-kit receptor and its ligand (stem cell factor), 15 expressed both proteins indicating the possibility that this autocrine stimulation feedback loop is a factor in the growth of some ovarian cancers. This study demonstrates that PDGFR alpha, PDGFR beta, and c-kit are expressed in a high percentage of epithelial ovarian cancers suggesting that tyrosine kinase inhibitors may be useful in the treatment of these tumors. PMID- 15791569 TI - Assessment of the HER2 status in breast cancer by fluorescence in situ hybridization: a technical review with interpretive guidelines. AB - Diagnostic assays for HER2 in breast cancer provide important prognostic information and independently help guide management by identifying patients who are the most likely to benefit from Herceptin-targeted therapy. The biological events underlying HER2 -driven breast cancer that can be assessed in routine clinical specimens include the evaluation of gene amplification by fluorescence in situ hybridization (FISH), enhanced messenger RNA expression by real-time polymerase chain reaction, and the assessment of protein overexpression at the tumor cell membrane by immunohistochemistry (IHC). Immunohistochemistry and FISH methodologies have the advantage of being morphologically driven, allowing for correlations between HER2 expression and morphologic features. However, each has important advantages and disadvantages, which are discussed in detail. Although immunohistochemistry is familiar and readily accommodated in most surgical pathology laboratories, increasing demands for FISH testing in the clinical setting will require greater familiarity with the technical aspects of FISH assays and their interpretation by the greater laboratory community. In this review, we provide an overview of FISH testing for HER2 in breast cancer, with an emphasis on technical considerations, interpretive guidelines, scoring criteria, and quality control. The development of automated platforms for hybridization, image analysis for signal enumeration, and experience with FISH interpretation should broaden the availability of this technology for clinical diagnostic testing. PMID- 15791570 TI - C-kit expression in renal oncocytomas and chromophobe renal cell carcinomas. AB - C- kit encodes the membrane-bound tyrosine kinase KIT, whose expression has been identified in several types of human neoplasms. Recently, KIT has been reported to be a marker for chromophobe renal cell carcinoma (RCC) and renal angiomyolipoma. However, expression of this molecule has not been adequately studied in other renal tumors, particularly oncocytoma, which may morphologically resemble chromophobe RCC. In this study, we analyzed c- kit messenger RNA (mRNA) levels in 17 chromophobe RCCs and 20 renal oncocytomas obtained from complementary DNA (cDNA) microarrays. Furthermore, comprehensive immunohistochemical analysis of KIT protein using a monoclonal antibody was performed in 226 renal tumors including chromophobe RCC (n=40), oncocytoma (n=41), clear-cell RCC (n=40), renal angiomyolipoma (n=29), and papillary RCC (n=21) on tissue microarrays (TMAs) and was compared with immunostaining results from 25 chromophobe RCCs and 30 oncocytomas using standard sections. The staining intensity was semiquantitatively graded on a 3-tier scoring system. All chromophobe RCCs and oncocytomas showed significant overexpression of c- kit mRNA. The average increase of mRNA compared with normal kidney tissue was 7.4 fold for chromophobe RCCs and 7.4-fold for oncocytomas. Immunohistochemical expression of KIT was found in most chromophobe RCCs (95% in TMAs and 96% in conventional sections) and oncocytomas (88% in TMAs and 100% in conventional sections) but was infrequently observed in renal angiomyolipomas (17%), papillary RCCs (5%), and clear-cell RCCs (3%). Furthermore, the average KIT immunoreactivity in TMAs was stronger in chromophobe RCC (1.93) and oncocytoma (2.07) than in other subtypes of renal tumors tested, including angiomyolipomas (0.17), papillary RCCs (0.05), and clear-cell RCCs (0.03). In conclusion, we found a significant elevation of c- kit mRNA by cDNA expression microarrays and overexpression of KIT protein by immunohistochemistry not only in chromophobe RCCs but also in oncocytomas. In contrast, immunohistochemical expression of KIT was not detected in most other types of renal cell tumors evaluated. The differential expression of c- kit in these renal tumors may have diagnostic and therapeutic implications. PMID- 15791571 TI - Islands of squamous epithelium and their surrounding mucosa in columnar-lined esophagus: a pathognomonic feature of Barrett's esophagus? AB - We examined 23 patients with columnar-lined esophagus (2 with long segment and 21 with short segment) endoscopically. After staining with Lugol's iodine solution, squamous islands were identified in the columnar mucosa in 18 (78%) of the 23 patients. When biopsy specimens were obtained from these islands, esophageal glands proper or their ducts were seen in 12 (67%) of the 18 cases. Because the identification of esophageal glands proper is a definite indicator that a piece of biopsy tissue is of esophageal origin, a diagnosis of columnar-lined esophagus could be made purely on the basis of the histological findings in these biopsy specimens of squamous islands. This was the case in 12 (52%) of the 23 patients examined in this study. Staining with Lugol's iodine solution, with subsequent biopsy of stained squamous islands, may assist in the diagnosis of short-segment columnar-lined esophagus. We also conclude, on the basis of our study, that columnar metaplasia of the esophagus cannot develop by direct outgrowth of epithelium from the ducts of esophageal glands proper. In addition, intestinal metaplasia was not always observed in the columnar mucosa around the duct orifices. From a practical point of view, biopsy specimens from columnar-lined mucosa of the esophagus do not always show intestinal metaplasia (specialized intestinal metaplasia). PMID- 15791572 TI - CK20 and CK7 protein expression in colorectal cancer: demonstration of the utility of a population-based tissue microarray. AB - The ability to use archival tissue to test externally valid hypotheses of carcinogenesis is dependent on the availability of population-based samples of cancer tissue. Tissue microarrays (TMAs) provide an efficient format for developing population-based samples of tissue. A TMA was constructed consisting of archival tissue from patients diagnosed with invasive colorectal cancer in the state of Hawaii in 1995. The population representativeness of the TMA was evaluated by comparing patient and clinical characteristics of TMA cases to that of all cases of colorectal carcinoma diagnosed statewide in 1995. Cytokeratin 20 (CK20) and cytokeratin 7 (CK7) immunohistochemistry was used to validate the utility of the TMA, and the expression of these proteins was correlated with patient and tumor characteristics. The TMA comprised tissue specimens from 286 patients representing 47% of all invasive cases diagnosed statewide in 1995. TMA cases were comparable to all invasive colorectal cases statewide with respect to age, sex, race/ethnicity, anatomic site, and survival. There were some differences between TMA cases and all cases with respect to tumor stage, histological classification, and treatment. There were significant differences in the relative expression of CK20 and CK7 proteins between malignant and normal tissues and by tumor stage. Advanced cancers were more likely to have CK20+/cytokeratin 7+ (CK7+) profiles than early-stage cancers, which were predominantly CK20+/cytokeratin 7- (CK7-). CK7+ expression was not correlated with anatomic location of carcinomas. This well-characterized TMA offers a powerful tool for testing hypotheses regarding colorectal carcinogenesis, including the identification of potential markers of neoplastic development and progression. PMID- 15791573 TI - Lymphomatosis cerebri as a cause of white matter dementia. AB - Primary central nervous system lymphoma most often presents as a solitary, isolated lesion in immunocompetent patients. Rarely, the disease presents as a diffuse, infiltrating condition without formation of a cohesive mass, a pattern called lymphomatosis cerebri. We present 3 immunocompetent individuals who developed rapidly progressive dementia. Magnetic resonance imaging features mimicked other disorders of white matter and prompted preoperative diagnoses of Binswanger's disease (subcortical ischemic vascular dementia), unknown leukoencephalopathy, viral infection, or infiltrating glioma. Neuropathologic examination at biopsy (Poon T, Matoso I, Tchertkoff V, Weitzner I Jr, Gade M. CT features of primary cerebral lymphoma in AIDS and non-AIDS patients. J Comput Assist Tomogr . 1989;13:6-9) and autopsy (Schwaighofer BW, Hesselink JR, Press GA, Wolf RL, Healy ME, Berthoty DP. Primary intracranial CNS lymphoma: MR manifestations. Am J Neuroradiol . 1993;10:725-9) demonstrated nonnecrotic, diffusely infiltrating, large-cell B-cell lymphoma of white matter, with relative sparing of gray matter, and without significant leptomeningeal involvement or bulky periventricular disease at autopsy. Microglial and astrocytic reactions, but only subtle myelin pallor, were evident as individual tumor cells permeated the entire brain and spinal cord, albeit with considerable variation in cell density. Individual tumor cells could be identified from the optic nerve to spinal cord, documenting the "whole-brain" nature of the disease. CD20 immunostaining was necessary to fully appreciate the extent of individual lymphoma cell percolation through the white matter. The neurobehavioral deficits manifested by these patients demonstrate that lymphomatosis cerebri is an additional neoplastic cause of white matter dementia and can be added to the growing list of disorders responsible for this syndrome. PMID- 15791574 TI - Granular cell tumor of the penis: clinicopathologic evaluation of 9 cases. AB - The occurrence of granular cell tumor (GCT) in penile tissue is very rare, with only 9 examples reported to date in the English-language literature. Herein, we describe the clinicopathologic and immunohistochemical findings in 9 additional cases. The patients ranged in age from 20 to 60 years (mean, 42 years; median, 40 years) at time of diagnosis. All penile tumors were solitary and arose in the dermis of the penile shaft (n=4), prepuce (n=3), and corona (n=2). A patient had a history of multiple cutaneous GCTs. Duration of symptoms before surgery ranged from 5 days to 2 years with the presence of an asymptomatic nodule representing the most common tumor-related complaint (n=8). The lesions ranged in size from 0.6 to 2.5 cm (mean, 1.5 cm; median, 1.5 cm). Microscopically, the tumors were moderate to highly cellular and were composed of oval to polygonal-shaped cells with abundant coarsely granular eosinophilic cytoplasm. Tumor cells grew in infiltrating nests, cords, and trabeculae and showed neural (n=2) and vessel wall (n=1) invasion or formed a relatively well-marginated solid nodule. Bland cytological features with only rare cells showing nucleomegaly (n=7) or spindling (n=3) were exhibited by 8 tumors. A tumor demonstrated diffuse nuclear atypia and was classified as "atypical." Mitotic activity ranged from 0 to 8 mitoses (mean, 1.4 mitoses) per 50 high-powered fields with no atypical division figures identified. All tumors tested showed moderate to strong immunohistochemical expression of S100 protein (n=6) and low-affinity nerve growth factor receptor (n=5), which was useful for detecting small deposits of tumor and helpful in evaluating surgical margins. Focal tumor cell immunoreactivity was observed for calretinin (4/6 cases) and glial fibrillary acidic protein (1/6 cases). All patients underwent simple (local) excision of their tumor. Complete follow-up data (mean, 21 years; interval range, 0.5-28 years) were available for 6 patients. No patient experienced recurrence or metastatic spread of tumor although surgical margins were microscopically involved by tumor in 5 cases. Benign GCT involving superficial soft tissue of the penis can be adequately managed by a simple excision. Patients with microscopically involved surgical margins can be clinically followed without immediate additional surgery. PMID- 15791575 TI - Glomus tumor of renal pelvis: a case report and review of the literature. AB - Glomus tumors are uncommon benign perivascular neoplasms that have rarely been described outside of their usual peripheral soft tissue sites. We report a unique case of glomus tumor of the renal pelvis in a 53-year-old woman who presented with microscopic hematuria associated with obstruction of the ureteropelvic junction and marked hydronephrosis. At initial gross examination, the tumor mimicked a urothelial carcinoma. PMID- 15791576 TI - Immunohistochemical, in situ hybridization, and ultrastructural localization of SARS-associated coronavirus in lung of a fatal case of severe acute respiratory syndrome in Taiwan. AB - This article describes the pathological studies of fatal severe acute respiratory syndrome (SARS) in a 73-year-old man during an outbreak of SARS in Taiwan, 2003. Eight days before onset of symptoms, he visited a municipal hospital that was later identified as the epicenter of a large outbreak of SARS. On admission to National Taiwan University Hospital in Taipei, the patient experienced chest tightness, progressive dyspnea, and low-grade fever. His condition rapidly deteriorated with increasing respiratory difficulty, and he died 7 days after admission. The most prominent histopathologic finding was diffuse alveolar damage of the lung. Immunohistochemical and in situ hybridization assays demonstrated evidence of SARS-associated coronavirus (SARS-CoV) infection in various respiratory epithelial cells, predominantly type II pneumocytes, and in alveolar macrophages in the lung. Electron microscopic examination also revealed coronavirus particles in the pneumocytes, and their identity was confirmed as SARS-CoV by immunogold labeling electron microscopy. This report is the first to describe the cellular localization of SARS-CoV in human lung tissue by using a combination of immunohistochemistry, double-stain immunohistochemistry, in situ hybridization, electron microscopy, and immunogold labeling electron microscopy. These techniques represent valuable laboratory diagnostic modalities and provide insights into the pathogenesis of this emerging infection. PMID- 15791577 TI - Breast epithelial cells in dermal angiolymphatic spaces: a manifestation of benign mechanical transport. AB - Evidence suggesting that breast epithelial cells may reach axillary lymph nodes by benign mechanical transport (BMT), rather than metastatic means, has been recently reported. We report a case of a patient with ductal carcinoma in situ of the breast, who had displaced epithelial elements in the mastectomy specimen, dermal angiolymphatic spaces, and a sentinel lymph node. We attribute the epithelial aggregates in the dermal angiolymphatic spaces and a single cell in the sentinel lymph node to BMT, based on the clinicopathological findings of the case. We, therefore, suggest that the effects of BMT be considered in the differential diagnosis of epithelial aggregates in dermal angiolymphatic spaces in the appropriate clinicopathological setting. PMID- 15791578 TI - Immunocytochemical localization of MAPKAPK-2 and Hsp25 in the rat temporomandibular joint. AB - One series of our research has shown an intense expression of immunoreaction for heat shock protein 25 (Hsp25) in various cellular elements in the rat temporomandibular joint (TMJ). This protein is the major substrate of mitogen activated protein kinase-activated protein kinase-2 (MAPKAPK-2), which mediates an intracellular stress-activated signaling pathway to stimulate cytosolic actin reorganization under various stresses. The present study was undertaken to examine the localization of MAPKAPK-2 in the rat TMJ by immunocytochemical techniques. Furthermore, confocal microscopy with double staining was employed to demonstrate the colocalization of MAPKAPK-2 and Hsp25. Immunocytochemistry for MAPKAPK-2 showed an intense immunoreaction in the cytoplasm of the synovial lining cells, the endothelial cells, and the fibroblasts in the synovial membrane of the rat TMJ. Double immunostaining under a confocal microscope succeeded in demonstrating the colocalization of MAPKAPK-2 and Hsp25 immunoreactions in the cytoplasm of fibroblastic type B synoviocytes in the TMJ. On the other hand, the macrophage-like type A-cells expressed MAPKAPK-2 immunoreactions but lacked Hsp25 immunoreactivity. The cells in the articular disk and the chondrocytes in the maturative and hypertrophic layer of the mandibular cartilage also showed intense immunoreactions for MAPKAPK-2 and Hsp25. In addition to cytoplasmic localization, MAPKAPK-2 immunoreactions were found in the nucleus of some synovial lining cells, cells in the articular disk, and chondrocytes. Current observations imply the presence of the phosphorylation of Hsp25 via activated MAPKAPK-2 in the cytoplasm. MAPKAPK-2 and Hsp25 possibly participate in the induction of cytoskeletal changes to the various cellular elements in rat TMJ under normal conditions. PMID- 15791579 TI - Senescence-associated HSP60 expression in normal human skin fibroblasts. AB - Normal mammalian fibroblasts cultured in vitro undergo a limited number of divisions before entering a senescent phase in which they can be maintained for long periods but cannot be induced to divide. Senescent cells become unresponsive to growth-promoting signals and exhibit senescent cell morphology with flattened and enlarged cell shape. Several chaperones have a direct effect on cellular senescence. HSP60 has been largely studied in our laboratories and it has been associated with uncontrolled cell proliferation in tumor cells. Since senescence is firmly regulated during cell cycle progression, we wanted to investigate HSP60 protein level during cellular senescence. Our data show that HSP60 increases during the initial stage of senescence and that it is localized in cellular compartments, resembling mitochondria. An increase in HSP60 protein amount is associated with a cell cycle slow-down and it may have a role in cell cycle progression. PMID- 15791580 TI - Evidence for frequency-dependent arterial damage in vibrated rat tails. AB - The effects of single 4-hr bouts of continuous 30, 60, 120, and 800 Hz tail vibration (49 m/sec2, root mean squared) were compared to assess frequency amplitude-related structural damage of the ventral caudal artery. Amplitudes were 3.9, 0.98, 0.24, and 0.0055 mm, respectively. Vibrated, sham-vibrated, and normal arteries were processed for light and electron microscopy. The Curry rat tail model of hand-arm vibration (Curry et al. Muscle Nerve 2002;25:527-534) proved well-suited for testing multiple frequencies. NFATc3 immunostaining, an early marker of cell damage, increased in smooth muscle and endothelial cells after 30, 60, and 120 Hz but not 800 Hz. Increased vacuolization, which is indicative of smooth muscle contraction, occurred for all frequencies except 800 Hz. Vacuoles increased in both endothelial and smooth muscle cells after 60 and 120 Hz. Only 30 Hz showed pronounced smooth muscle cell vacuolization along the internal and external elastic membranes, suggesting stretch-mediated contraction from the large amplitude shear stress. Discontinuities in toluidine blue staining of the internal elastic membrane (IEM) increased for all frequencies, indicating vibration-induced structural weakening of this structure. Patches of missing IEM and overlying endothelium occurred in approximately 5% of arteries after 60, 120, and 800 Hz. The pattern of damage after 800 Hz suggests that the IEM is disrupted because it resonates at this frequency. Vibration acceleration stress and smooth muscle contraction appear to be the major contributors to arterial damage. The pattern of vibration-induced arterial damage of smooth muscle and endothelial cells is frequency-amplitude-dependent. PMID- 15791581 TI - Distribution of extracellular signal-regulated kinase 1- and 2-activated neurons in the rat periaqueductal gray matter after noxious stimulation. AB - The periaqueductal gray matter (PAG), the midbrain region made up of neuronal columns encircling the cerebral aqueduct, plays a key role in nociception. As the extracellular signal-regulated kinases (ERKs) 1 and 2 are activated after noxious stimulation, we analyzed the distribution of ERK-activated neurons in the PAG after visceral noxious stimulation. Ether- and urethane-anesthetized rats received an intraperitoneal injection of acetic acid or were left untreated and were perfused after 2 hr. Serial sections immunoreacted with an antibody selective for the activated ERKs. Significant ERK activation occurred only in the ether-anesthetized noxious stimulated rats. In these rats, we evaluated the number of ERK-activated neurons and their density as the ratio of the number of immunolabeled neurons to the extension of the region where they were located. ERK activated neurons were more numerous in the lateral (LPAG) and ventrolateral (VLPAG) columns, but without significant differences. No ERK activation was seen in neurons of the most rostral PAG. The ERK-activated neurons were significantly denser at the intermediate level of the PAG. At the caudal level, they were denser in the LPAG and VLPAG columns, and in the DPAG column at the intermediate and rostral level. These findings suggest that noxious stimulation activates ERKs in neurons involved in the different functional activities related to nociception, overlapping in the PAG columns, and strengthens the role of PAG in integration. PMID- 15791582 TI - Ultrastructural and immunocytochemical studies of the viscacha (Lagostomus maximus maximus) pituitary pars tuberalis. AB - The hypophyseal pars tuberalis (PT) has been the focus of numerous studies attempting to understand its physiological role in the reproductive regulation and modulation by the neuroendocrine system. Ultrastructural studies of the PT in a number of species have shown that it consists of a well-developed hypophyseal area with important secretory activity, demonstrated by the abundance of secretory granules in the cytoplasm and the marked blood irrigation. This article describes ultrastructural and immunocytochemical aspects of the PT in viscachas captured in their habitat. The cell types identified were PT-specific cells, agranulated cells, and Folliculostellate cells. PT-specific cells are divided into type I and II. Type I cells have cytoplasms with secretory granules of 150 500 nm diameter. The secretory granules of type II PT-specific cells are 65-200 nm in diameter. Both cellular types exhibit numerous nerve endings on the plasmatic membranes. Agranulated cells exhibit nuclei with lax chromatin, mitochondria, phagosomes, scarce Golgi complex, and rough endoplasmic reticulum. Folliculostellate cells exhibit an irregularly shaped and moderately condensed nucleus. All the described cellular types exhibit deposits of cytoplasmic glycogen. The immunocytochemical study revealed the presence of cells immunostained for LH-beta and FSH-beta in the PT caudal zone. ACTH was only detected in the zona tuberalis. No staining was observed with antiprolactin, anti TSH-beta, and anti-GH sera. Folliculostellate cells exhibited staining with anti S-100. The results demonstrate that the viscacha PT is a hypophyseal zone with specific cellular types, which exhibits evident secretory activity. The presence of nerve endings suggests neural control of the function of PT cells. PMID- 15791583 TI - Synthesis of alveolar bone Sharpey's fibers during experimental tooth movement in the rat. AB - There is little information concerning the effects of tooth movement on the relative synthesis of bone matrix and Sharpey's fiber collagenous proteins. The purpose of this study was to investigate this situation using radioautographic techniques. The maxillary first molar tooth in rats was tipped toward the midline using an appliance and the animals were injected with 3H-proline after 3 days and sacrificed 24 hr later. Maxillae were sectioned and silver grain proportional areas (grain density/5,000 microm2) evaluated over Sharpey's fibers and adjacent alveolar bone matrix using computerized densitometry and histomorphometric techniques. These data were compared to a group of untreated animals by Fisher's exact test. At depository surfaces of experimental tissues, the silver grain proportional area over bone matrix was significantly greater than over Sharpey's fibers (P<0.05) and control bone matrix (P<0.01). The silver grain proportional area over Sharpey's fibers was not different between the groups. At resorptive surfaces, the silver grain proportional area over both bone matrix and Sharpey's fibers was significantly greater in experimental tissues compared to controls (P<0.01). Thus, movements of adjacent teeth affect both the quantity and ratios of collagenous protein incorporation into Sharpey's fibers and adjacent alveolar bone, which is dependent on the intensity and characteristics of the force. PMID- 15791584 TI - Structure and biomechanical properties of the trachea of the striped dolphin Stenella coeruleoalba: evidence for evolutionary adaptations to diving. AB - This study analyzes the structure and mechanical properties of the trachea of the striped dolphin Stenella coeruleoalba, one of the most common cetacean species. The cetacean trachea is made up of closed or semiclosed cartilaginous rings without a paries membranaceus. Our results indicate that the inner lining of the trachea contains erectile tissue in which several venous lacunae permeate the mucosa. We also observed and described the presence of peripheral neurons containing nitric oxide along the rim of the venous lacunae. Data obtained from compression and tensile tests and comparison with the pig and goat tracheas indicate a higher stiffness and a different, higher breaking point for the dolphin trachea. On the whole, our data suggest that the trachea of the striped dolphin possesses structural properties that allow rapid filling with blood, possibly in relation to dive activities, and also allow modifications due to increased pressure and immediate return to the original shape without risks of permanent bending or rupture, as would happen in a terrestrial mammal. As the organ undergoes intense pressure difference during descent to optimal foraging depth and subsequent rapid ascent to surface, especially in deep dives of hundreds of meters, the specific structural and biomechanical peculiarities of the trachea of the striped dolphin may represent an evolutionary adaptation to life in the water and to diving. PMID- 15791585 TI - Development of the subchondral bone layer of the medial coronoid process of the canine ulna. AB - The medial coronoid process (MCP) of the ulna takes part in the weight-bearing function of the elbow in quadrupedal animals. In this study, the timing of development of a solid subchondral bone layer (SBL) of the MCP in the dog is investigated, as this might be important in the pathogenesis of the fractured medial coronoid process, a common disease in young dogs of larger breeds. The SBL is considered to make an important contribution to the strength of the MCP. In this study, the SBL is visualized at the humeral articular side (H-side) and in the radial notch (RN) via three-dimensional reconstructions of micro-CT scans (34 mum voxel size) in nine young golden retrievers. After micro-CT scanning, the area was investigated histologically. Gradually, the appearance of the SBL on the H-side changes from a trabecular aspect to an even surface with gaps and finally to a completely even surface. The surface in the RN is still rough at 24 weeks after birth, although some consolidation has occurred. Initially, the enchondral ossification activity, as observed in the histological sections, is high, but later, when the intertrabecular spaces are filled in with calcified cartilage and bone, activity is less evident. Some vessels penetrated the SBL, but it is unclear if they account for all the gaps in the surface seen in micro-CT. In addition, the formation of a cortical structure of the proximal ulnar shaft could be visualized. The bony cortex is already even at the mediocaudal side of the proximal ulna 4 weeks after birth, but remains trabecular at the dorsal side until 6 weeks later. We hypothesize that the observed differences in the formation of an even SBL or cortex can be explained by mechanical factors. A smooth cortical layer has an even thickness and will be stronger than a cortex with varying thickness. In the MCP, compressive forces exerted by the humerus may be responsible for the early smoothening on the H-side. In the proximal ulna, the resistance to bending in the sagittal plane will depend mainly on the caudal surface of the ulna (and dorsal surface of the radius), surfaces furthest away from the neutral axis. At least the caudal ulna smoothened first, thus providing stiffness against bending at an early age. PMID- 15791586 TI - Small GTPase RhoA is required for ooplasmic segregation and spindle rotation, but not for spindle organization and chromosome separation during mouse oocyte maturation, fertilization, and early cleavage. AB - RhoA, a small GTPase, plays versatile roles in many aspects of cell function such as stress fiber formation, cytokinesis, and cell polarization. In this study, we investigated the subcellular localization of RhoA and its possible roles during oocyte maturation and fertilization. RhoA was localized in the cytoplasm of eggs from the germinal vesicle (GV) stage to 2-cell stage, especially concentrating in the midbody of telophase spindle when oocyte extruded PB1 and PB2. The RhoA kinases (ROCKs) specific inhibitor Y-27632 blocked GV breakdown (GVBD) and first polar body extrusion, but did not affect apparatus formation and anaphase/telophase I entry. Anti-RhoA antibody microinjection into the oocytes showed similar results. RhoA inhibitor caused abnormal organization of microfilaments, failure of spindle rotation, PB2 extrusion as well as cleavage furrow formation, while sister chromatid separation was not affected. Microinjection of RhoA antibody also blocked PB2 emission. Our findings indicate that RhoA, by regulating microfilament organization, regulates several important events including GVBD, polar body emission, spindle rotation, and cleavage. PMID- 15791587 TI - Ejaculates from the common marmoset (Callithrix jacchus) contain semenogelin and beta-microseminoprotein but not prostate-specific antigen. AB - Human seminal plasma contains high concentrations of prostatic acid phosphatase (PAP), prostate-specific antigen (PSA), beta-microseminoprotein (MSP), semenogelin I (SgI), and semenogelin II (SgII), whereas only PAP and MSP are present in rodents. In order to gain a better understanding of the evolution and function of semen proteins, we have studied ejaculates from the common marmoset (Callithrix jacchus)-a New World monkey. Semen samples were analyzed with SDS PAGE, Western blotting, and isoelectric focusing. Under reducing conditions the dominating protein components appear as heterogeneous material of 55-70 kDa and distinct protein bands of 85, 17, 16, and 15 kDa. The heterogeneous material contains glycosylated material detected by an antiserum recognizing both human SgI and SgII. Southern blotting indicates that the common marmoset has genes for both SgI and SgII. There are several marmoset MSP genes, but the strong immunoreactivity against one 15 kDa semen component with pI 7.3 suggests preferential expression of one gene in the prostate. Expression of two other genes cannot be excluded as indicated by weak reaction to isoforms with pI 6.6 and 4.9. Unexpectedly, PSA was not detected by either immunological methods or activity measurements. This is in agreement with results from Southern blotting suggesting that the common marmoset might not have a PSA gene. Thus, in this study we have shown that semen coagulum proteins are present in marmoset seminal plasma, but the lack of PSA precludes a similar liquefaction as of human semen. PMID- 15791588 TI - Concomitant anomalies in 100 children with unilateral multicystic kidney. AB - OBJECTIVES: To determine the incidence and type of associated urogenital anomalies in children with a unilateral multicystic kidney and to assess in children with nephrectomy the additional diagnostic value of cystoscopy and, in girls, of colposcopy. METHODS: This was a follow-up study of 100 fetuses with antenatally detected unilateral multicystic kidneys. After ultrasound confirmation of the diagnosis within a few days after birth voiding cystourethrography and isotope scan were performed in 83 of the surviving children to exclude vesicoureteral reflux and to establish renal function. Eighty one children underwent nephrectomy and, prior to surgery, all underwent cystoscopy and girls also underwent colposcopy. RESULTS: Seventy-five children had one or more additional urogenital anomalies: 39 had anomalies of the contralateral kidney, 40 had anomalies of the ipsilateral kidney and 30 had one or more anomalies of the lower urogenital tract. With cystoscopy 54 anomalies of the genitourinary tract were detected in 48 children and with colposcopy three anomalies were detected in 35 girls. Eighty-one children had a nephrectomy or heminephrectomy and 33 of them needed other urological intervention. Thirteen fetuses died (mostly from agenesis of the contralateral kidney) and six infants had no surgery at all. CONCLUSION: Children with a unilateral multicystic kidney are at considerable risk of having other urogenital anomalies. When cystoscopy and colposcopy are added to routine investigations the rate of detection of anomalies is 75%, twice that reported in the literature. PMID- 15791589 TI - Asymmetric division of spindle microtubules and microfilaments during bovine meiosis from metaphase I to metaphase III. AB - The kinetics of spindle and chromosomes during bovine oocyte meiosis from meiosis I to meiosis III is described. The results of this study showed that (1) oocytes began to extrude the first polar body (Pb1) at the early anaphase I stage and the Pb1 totally separated from the mother cell only when oocytes reach the MII stage; (2) the morphology of the spindle changed from barrel-shaped at the metaphase stage to cylinder-shaped at early anaphase, and then to a thin, long triangle shaped cone at late anaphase and telophase stages; (3) chromosome morphology went from an individual visible stage at metaphase to a less defined chromatin state during anaphase and telophase stages, and then back to visible individual chromosomes at the next metaphase; (4) chromatin that connected with the floor of the cone became the polar bodies and expelled, and almost all of the microtubules (MTs) and microfilaments (MFs) composing the spindles moved towards and contributed to the polar bodies; and (5) the size of the metaphase I (MI) spindle was larger than the metaphase II (MII) and metaphase III (MIII) spindles. The MII spindle, however, is more barrel-shaped than the MI spindle. This study suggests that spindle MTs and MFs during bovine oocyte meiosis are asymmetrically divided into the polar bodies. PMID- 15791590 TI - Configurations of germinal vesicle (GV) chromatin in the goat differ from those of other species. AB - Configuration of germinal vesicle (GV) chromatin has been studied and found correlated with the developmental competence of oocytes in several mammalian species. A common feature in the configuration of GV chromatin in the species studied so far is that the diffuse chromatin (the so called "NSN" pattern) condenses into a perinucleolar ring (the so called "SN" configuration) with follicular growth. However, no study has been published on the configuration of GV chromatin in the goat. Nor is it known whether the perinucleolar ring of condensed chromatin (CC) in an oocyte represents a step toward final maturation or atresia. Changes in configurations of GV chromatin and RNA synthesis during goat oocyte growth, atresia and maturation in vivo and in vitro were investigated in this study. Based on both the size of nucleoli and the degree of chromatin condensation, the GV chromatin of goat oocytes was classified into GV1 characterized by large nucleoli and diffuse chromatin, GV2 with medium-sized nucleoli and condensed net-like (GV2n) or clumped (GV2c) chromatin, GV3 with small nucleoli and net-like (GV3n) or clumped (GV3c) chromatin, and GV4 with no nucleolus but clumped chromatin. The results showed that (i) the configurations of GV chromatin in the goat differ from those of other species in that the chromatin did not condense into a perinucleolar ring; (ii) most of the goat oocytes are synchronized at the GV3n configuration before GVBD; (iii) the GVn pattern might represent a healthy state, but the GVc an atretic state; (iv) in both goats and mice, the GC-specific (Chromomycin A3, CMA3) and the AT-specific (Hoechst 33342) fluorochromes followed the same pattern of distribution in GV chromatin; (v) the nucleolar size decreased significantly with oocyte growth and maturation in vivo and in vitro; and (vi) goat oocytes began GVBD at 8 hr and had completed it by 20 hr after onset of estrus. The peculiar configuration of GV chromatin of goat oocytes can be a useful model for studies of morphological and functional changes of different nuclear compartments during the cell cycle and cell differentiation, and the functional differentiation between GV3n and GV3c might be used for reference to the question whether the "SN" configuration in other species inclines toward ovulation or atresia. PMID- 15791591 TI - Simulation of intrafollicular conditions prevents GVBD in bovine oocytes: a better alternative to affect their developmental capacity after two-step culture. AB - To increase the developmental competence of bovine oocytes isolated from small, medium, and large follicles (2-3, 3-4, and 4-6 mm in diameter, respectively), we tried to modify the conditions for their in vitro culture. The first step involved conditions maintaining at least for 48 hr a reversible inhibition of the germinal vesicle breakdown (GVBD) and the second step stimulated the resumption of meiosis and completion of nuclear and cytoplasmic maturation during the subsequent 20-22 hr of culture. The effectiveness of this model depended mainly on the medium composition (reduced NaHCO3, substitution of serum with serum albumin, addition of antioxidants (curcumin), increased viscosity by agar, the reduction of oxygen concentration (within 6%-8%), the reduction of the proportion between the number of cumulus-oocyte complexes (COCs), and the reduction of the amount of a medium (within 6-7 mul per COC) to amplify the GVBD-inhibitory effect of oocyte surrounding granulosa cells. The COCs were incubated in clumps of 6-7 COCs. The effectiveness and reversibility of GVBD inhibition depended also on the duration of COCs isolation. The full reversibility of the GV block was controlled morphologically and also by measuring histone H1 and MAP kinase activities. The two-step versus one-step (24 hr) maturation technique was evaluated by the percentage of total and hatched blastocysts at day 9. When compared with one-step maturation, the two-step culture showed a slightly increased proportion of total and hatched blastocysts developed from growing follicles, mainly from the smallest category (13.9% vs. 7.1% and 9.2% vs. 3.3% for total blastocysts and hatched, respectively). However, the two-step culture of oocytes from large regressing follicles substantially reduced the blastocyst yield (9.7% vs. 39.1% and 4.9% vs. 26.7% for total blastocysts and hatched, respectively). The transfer of ten blastocysts (developed after two-step culture) to ten recipients resulted in seven pregnancies. PMID- 15791592 TI - Role of the sperm proteasome during fertilization and gamete interaction in the mouse. AB - In this work, we have investigated the role of the sperm proteasome during in vitro fertilization (IVF) and gamete interaction in the mouse. Proteasome activity was measured in extract and intact sperm using a specific substrate. In addition, sperm were treated with specific proteasome inhibitors and evaluated during IVF, binding to the zona pellucida, and progesterone- and zona pellucida induced acrosome reactions. In other experiments, sperm membrane proteins were obtained resuspending them in Triton X-114, shaking vigorously and let standing by 4 hr. Soluble sperm proteins were partitioned in the aqueous phase and sperm membrane proteins in the detergent phase. In both phases, proteasome activity was measured. Labeling of cell surface sperm proteins was carried out with the cell impermeable NHS-LC biotin, extracted with Triton X-114, and mixing with avidin agarose beads. Nonpermeabilized sperm were incubated with an anti-proteasome monoclonal antibody and evaluated by indirect immunofluorescence. The results indicate that sperm extracts as well as intact sperm had proteasome activity; the sperm proteasome was involved in IVF, specifically during sperm-zona pellucida binding and the acrosome reaction; soluble sperm membrane proteins exhibited proteasome activity; biotin experiments indicated the presence of proteasomes on the sperm surface, which was corroborated by indirect immunofluorescence experiments. All these observations indicate that the mouse sperm proteasome participates in the binding to the zona pellucida and the acrosome reaction and that there is a pool of proteasomes located on the sperm head. PMID- 15791593 TI - Experimental demonstration that mammalian oocytes are not selective towards X- or Y-bearing sperm. AB - Mammalian oocytes are thought to be neutral as for X- or Y-bearing sperm selection is concerned, and penetration of an oocyte by an X- or a Y-bearing sperm is considered a random event. This assumption is mainly based on a posteriori evidences of a nearly equal sex ratio at birth, but it has never been experimentally demonstrated. We have designed a simple experiment, which allowed the penetration of an oocyte by more than one sperm and the further sexing by PCR of each single pronucleus present within the ooplasm. For the first time, we provide experimental evidence that mammalian oocytes do not play a selecting role since a single oocyte may be simultaneously fertilised by both X- and Y-bearing sperm. PMID- 15791594 TI - Transcriptional profiling using a novel cDNA array identifies differential gene expression during porcine embryo elongation. AB - A novel porcine cDNA array, containing 1,015 PCR products selected for embryonic expression, was used for transcriptional profiling of conceptuses at four stages of peri-implantation development. Total conceptus RNA from small spherical, large spherical, tubular, and filamentous stages was amplified, converted to cDNA, and hybridized to membranes. Initially, normalized signal intensities obtained using cDNA from total RNA or from amplified RNA were compared. Uniform distribution of P-values associated with t-tests conducted for each gene indicated no evidence that amplification introduced bias. Analysis of data obtained by using amplified targets and the novel array identified genes differentially expressed across stages. Such genes were identified by testing for significant stage effects in gene-specific mixed models. A total of nine genes were declared differentially expressed. Six of the nine genes had P-values less than 0.001, and a false discovery rate of approximately 17% was associated with this significance threshold. Two out of six genes were significant when using the Bonferroni method to control the probability of one or more false positives. The other three genes had P-values between 0.001 and 0.01 and exhibited differences greater than twofold between stages. All four genes selected for confirmation (steroidogenic acute regulatory protein, interleukin 1 beta, transforming growth factor beta 3, and thymosin beta 10) were shown to be differentially expressed by using quantitative real time RT-PCR. Our study shows that RNA amplification is useful for transcriptional profiling with limiting porcine embryonic RNA, and that this novel targeted array can detect differential gene expression during trophoblastic elongation. Finally, our results contribute to an increased understanding of the temporal patterns of expression of known genes controlling conceptus development, as well as identify novel genes also differentially regulated during implantation. PMID- 15791595 TI - Rho mediates cytokinesis and epiboly via ROCK in zebrafish. AB - To study the regulation of embryonic development by Rho, we microinjected Clostridium botulinum C3-exoenzyme (C3) into zebrafish embryos. We found that C3 inhibited cytokinesis during early cleavages. C3 inhibition appeared to be specific on RhoA, since the constitutively active RhoA could partially rescued the C3-induced defects. Distributions of actin and the cleavage furrow associated beta-catenin were disrupted by C3. Belbbistatin, a myosin II inhibitor, also caused blastomeres disintegration. It suggested that Rho mediates cytokinesis via cleavage furrow protein assembly and actomyosin ring constriction. Furthermore, C3 blocked cellular movements during epiboly and gastrulation as evident by the impairment on no tail and goosecoid expression in blastoderm front runner cells and the dorsal lip of blastopore, respectively. Y-27632, an antagonist of Rho associated kinase (ROK/ROCK), had the similar inhibitory effects on zebrafish development as the C3 treatments. Taken together, these results suggest that Rho mediates cleavage furrow protein assembly during cytokinesis and cellular migration during epiboly and gastrulation via a ROK/ROCK-dependent pathway. PMID- 15791596 TI - Baculovirus expressed C-terminal fragment of bonnet monkey (Macaca radiata) zona pellucida glycoprotein-3 inhibits ZP3-mediated induction of acrosomal exocytosis. AB - Zona pellucida glycoprotein-3 (ZP3) has been postulated as the primary sperm receptor in various mammalian species including bonnet monkey (Macaca radiata). However, information on the domain responsible for its binding to spermatozoa is inadequate. In the present study, bonnet monkey ZP3 (bmZP3), corresponding to amino acid (aa) residues 223-348 [bmZP3(223-348)] has been cloned and expressed using baculovirus expression system. SDS-PAGE and Western blot analysis of the purified renatured recombinant protein revealed it as a closely spaced doublet of approximately 25 kDa. Lectin-binding studies documented the presence of both O- as well as N-linked glycans. The biotinylated r-bmZP3(223-348) binds to the acrosomal region of the capacitated spermatozoa but fails to bind to the acrosome reacted spermatozoa as investigated by immunofluorescence studies. In ELISA, nonbiotinylated r-bmZP3(223-348) and baculovirus expressed r-bmZP3, devoid of signal sequence and transmembrane-like domain [r-bmZP3(23-348)] competitively inhibit its binding to the capacitated spermatozoa. Interestingly, binding of biotinylated r-bmZP3(23-348) to the capacitated sperm is also inhibited by nonbiotinylated r-bmZP3(223-348). In contrast to r-bmZP3(23-348), r-bmZP3(223 348) failed to induce acrosomal exocytosis in the capacitated sperm. Interestingly, it competitively inhibits the acrosomal exocytosis induced by r bmZP3(23-348). These studies, for the first time, identify a domain of ZP3 capable of binding to capacitated spermatozoa and inhibiting ZP3-mediated induction of acrosomal exocytosis furthering our understanding of mammalian fertilization. PMID- 15791597 TI - In vivo analysis of phagocytosis of apoptotic cells by testicular Sertoli cells. AB - Sertoli cells, a somatic cell type present within the seminiferous tubules of testes, are responsible for the phagocytic elimination of apoptotic spermatogenic cells. We here established an in vivo assay system that enables us to quantitatively analyze Sertoli cell phagocytosis of apoptotic cells in testes of live mice. Apoptotic cells were injected into the seminiferous tubules of spermatogenic cell-depleted mice, and the occurrence of phagocytosis by Sertoli cells was examined by histochemically analyzing testis sections or dispersed testicular cells. We reproducibly observed similar levels of phagocytosis in either examination, and the ratio of Sertoli cells that engulfed injected apoptotic cells was almost the same between the two examinations. These results indicated that a quantitative in vivo assay system was established using the seminiferous tubules of live mice as 'test tubes.' We then determined the requirements for Sertoli cell phagocytosis of apoptotic cells using this assay. For this purpose, apoptotic cells were injected together with various phagocytosis inhibitors, and the extent of phagocytosis by Sertoli cells was determined. The results revealed that Sertoli cells phagocytose apoptotic cells in a manner dependent on class B scavenger receptor type I (SR-BI) of Sertoli cells and phosphatidylserine exposed at the surface of target cells, as previously observed in vitro using primary cultures of dispersed rat testicular cells. Furthermore, the amount of SR-BI in Sertoli cells increased after injection of apoptotic cells into the seminiferous tubules, suggesting a positive feedback regulation of the expression of this phagocytosis receptor. PMID- 15791598 TI - Identification of rab12 as a vesicle-associated small GTPase highly expressed in Sertoli cells of rat testis. AB - We examined the expression and the localization of a small GTPase, rab12, in rat testis. Northern blot analysis showed that 2.3 kb transcript of rab12 was expressed in rat testis. RT-PCR analysis indicated constant expression of rab12 throughout testis development. Immunohistochemical studies revealed that rab12 protein was highly expressed in Sertoli cells in the seminiferous tubules, while both spermatogenic germ cells and interstitial cells exhibited faint or no immunosignal for rab12. The expression pattern of rab12 in Sertoli cells varied between the tubules: its immunostaining appeared as a wheel-like pattern at stage I approximately III and as a luminal staining pattern at stage IV approximately VI, whereas the immunostaining signals were only rudimentary detected at stage VIII and thereafter (approximately stage XIV). The diversified staining pattern of rab12 in the tubules seemed to reflect either the different shape of Sertoli cells during the cycle of the seminiferous epithelium or the variant expression levels of rab12 in Sertoli cells at each stage of the tubules. In cultured rat Sertoli cells and normal rat kidney (NRK) cells, rab12 was found to be associated with small vesicles distributed throughout the cytoplasm, but not with the Golgi apparatus. When overexpressed in NRK cells, rab12-associated small vesicles were not only distributed throughout the cytoplasm but also accumulated in the perinuclear cytoplasm around centrosome. We interrupt these data as a potential role of rab12 in acceleration of vesicular transport from the cell periphery to the perinuclear centrosome region. PMID- 15791599 TI - Simple and efficient production of mice derived from embryonic stem cells aggregated with tetraploid embryos. AB - Six newly derived hybrid mouse embryonic stem (ES) cell lines and two inbred ES cell lines were tested for their ability to produce completely ES cell-derived mice by aggregation of ES cells with tetraploid embryos. Forty-five ES cell tetraploid pups were generated from six hybrid ES cell lines and no pups from two inbred ES cell lines. These pups were found to have increased embryonic and placental weights than control mice. Twenty-two pups survived to adulthood and produced normal offsprings, and the other 23 pups died of several reasons including respiratory distress, abdomen ulcer-like symptoms, and foster failure. The 22 adult ES cell-tetraploid mice were completely ES cell-derived as judged by coat color and germline transmission, only two of them was found to have tetraploid component in liver, blood, and lung as analyzed by microsatellite loci. Our data suggested that genetic heterozygosity is a crucial factor for postnatal survival of ES cell-tetraploid mice, and tetraploid embryo aggregation using hybrid ES cells is a simple and efficient procedure for immediate generation of targeted mouse mutants from genetically modified ES cell clones, in contrast to the standard protocol, which involves the production of chimeras and several breeding steps. PMID- 15791600 TI - Fragmentation and development of preimplantation porcine embryos derived by parthenogenetic activation and nuclear transfer. AB - Fragmentation occurs during early developmental stages of electrically activated oocytes and nuclear transfer (NT) embryos. It might contribute to the low developmental rate of porcine NT embryos. The present study was conducted to investigate whether the addition of sugars such as sorbitol or sucrose suppresses fragmentation and supports the development of electrically activated oocytes and NT embryos. The activated oocytes were cultured in Porcine Zygote Medium-3 (PZM 3) supplemented with sorbitol or sucrose for 2 days after electric activation, and then cultured in the PZM-3 for the remaining 4 days. The osmolarities of PZM 3, PZM-3 supplemented with 0.05 or 0.1 M sorbitol, and PZM-3 with 0.05 M sucrose were 269 +/- 6.31, 316 +/- 3.13, 362 +/- 4.37, and 315 +/- 5.03 mOsm, respectively. When parthenogentically activated oocytes were cultured in PZM-3 supplemented with 0.05 M sorbitol or sucrose for the first 2 days and then cultured in PZM-3 without sugar, a significantly higher (P < 0.05) cleavage rate and blastocyst rate were observed. Interestingly, addition of sugar to PZM-3 for 2 days reduced the fragmentation rate compared to PZM-3 without sugar. In NT embryos, sugar addition into PZM-3 increased the fusion rate (84.2% +/- 6.07 vs. 95.1% +/- 2.52), cleavage rate (67.6% +/- 5.80 vs. 77.3% +/- 3.03), and developmental rate to the blastocyst stage (10.2% +/- 0.79 vs. 19.4% +/- 1.77). There was no significant difference between treatments for the number of the blastocysts. In addition the fragmentation rate was reduced compared to PZM-3 without sorbitol (26.1 +/- 4.30 vs. 14.5 +/- 1.74). In conclusion, increasing the osmolarity of PZM-3 through addition of either sorbitol or sucrose for 48 hr increased the cleavage and developmental rate to the blastocyst stage by reducing the fragmentation rate through increasing osmolarity. PMID- 15791601 TI - Lack of RNA-DNA oligonucleotide (chimeraplast) mutagenic activity in mouse embryos. AB - There are numerous reports of the use of RNA-DNA oligonucleotides (chimeraplasts) to correct point mutations in vitro and in vivo, including the human apolipoprotein E gene (ApoE). Despite the absence of selection for targeting, high efficiency conversion has been reported. Although mainly used to revert deleterious mutations for gene therapy applications, successful use of this approach would have the potential to greatly facilitate the production of defined mutations in mice and other species. We have attempted to create a point mutation in the mouse ApoE gene by microinjection of chimeraplast into the pronuclei of 1 cell mouse eggs. Following transfer of microinjected eggs we analysed 139 E12.5 embryos, but obtained no evidence for successful conversion. PMID- 15791602 TI - Expression of beta adrenergic receptors in mouse oocytes and preimplantation embryos. AB - Accumulating evidence indicates the role of endogenous catecholamines in mammalian embryogenesis. We searched public databases containing nucleotide sequences derived from mouse preimplantation cDNA libraries and found a partial sequence homology between a cDNA clone from mouse blastocysts and the mouse beta 2-adrenergic receptor sequence. No significant sequence homology was found for other mouse adrenergic and dopamine receptors. Using RT-PCR, we showed that beta 2-adrenoceptor is transcribed not only at blastocyst stage but also at earlier stages of preimplantation development as well as in oocytes. Moreover, we demonstrated that transcripts encoding both isoforms of the beta 3-adrenoceptor (beta 3a- and beta 3b-) are expressed in mouse oocytes and preimplantation embryos as well. We did not detect the beta 1-adrenoceptor transcript either in oocytes or in preimplantation embryos. Using an antibody against the mouse beta 2 adrenergic receptor, we showed that the receptor protein is expressed in oocytes and preimplantation embryos; in blastocysts, the immufluorescence labeling was stronger in the inner cell mass than in throphectodermal cells. The cell number of the in vitro cultured mouse preimplantation embryos exposed to isoproterenol (a potent beta adrenoceptor agonist) was lower than in control embryos, suggesting that activation of beta adrenergic receptors by appropriate agonist concentration can influence cell proliferation in mouse pre-implantation embryos. Thus, our results indicate that beta adrenergic receptors are expressed in mouse oocytes and preimplantation embryos and that ligands for the receptors can affect the mouse embryo even in the very early stages of development. PMID- 15791603 TI - A randomized crossover trial to examine administration techniques related to the Bristol female lower urinary tract symptom (BFLUTS) questionnaire. AB - AIMS: To investigate whether two methods of assessing lower urinary tract symptoms, interview-assisted standardized questionnaires, and self-completed standardized questionnaires, were comparable. METHODS: Women referred to a tertiary urogynecology urodynamic clinic with lower urinary tract symptoms were recruited. The psychometrically robust Bristol Female Lower Urinary Tract Symptoms (BFLUTS) questionnaire was used to assess urinary symptoms. A randomized crossover design for questionnaire completion was used: Group A-an initial interview-assisted questionnaire at the urodynamic clinic with a follow-up postal questionnaire; or Group B-an initial pre-out-patient postal questionnaire followed up by a second interview-assisted questionnaire whilst attending a scheduled urodynamic clinic. RESULTS: A total of 114 women were randomized for initial questionnaire administration method to achieve 72 completed sets (63%). Total scores were higher on self-completion compared to interview, this difference was not significant [t = -1.951, P > 0.05]. No item had perfect agreement between postal and interview methods and generally, interview-assisted questions had lower scores, indicating better health status. The items with the worst agreement (slight or moderate) were those concerned with sexual health, with women giving lower scores, exaggerating health on interview. For example bother of vaginal discomfort (kappa(w) = 0.09), compared to question assessing frequency (kappa(w) = 0.65). CONCLUSIONS: This data suggests women respond differently to the BFLUTS questionnaire when data is collected by self-completion in the woman's home compared to interviews. No individual question item had perfect agreement between the two methods of administration. Overall, individual question items were scored higher on self-completion than interview. PMID- 15791604 TI - Real-time image equalization for coronary X-ray angiography. AB - Coronary angiograms, which provide detailed images of contrast-filled coronary arteries, also show other large structures such as the diaphragm, spine and adjacent lung field. A real-time image processing method to attenuate these unwanted features is presented. Side-by-side comparisons of images selected from cine runs before and after processing show that the arteries in the processed images can be visualized more easily due to their higher contrast as other structures are made less prominent. It is also shown experimentally that this method allows more quantitative comparisons of the contrast of vessels in different parts of an image. PMID- 15791605 TI - The crab, the turkey and a malignant tale from the year of the rooster. PMID- 15791606 TI - Acupuncture for nocturnal enuresis in children: a systematic review and exploration of rationale. AB - OBJECTIVES: This review identified reports of acupuncture for childhood nocturnal enuresis, with the aim of ascertaining whether acupuncture is efficacious and or better than standard therapy for treating enuresis. MATERIALS AND METHODS: Studies of children of either gender <18 years of age who received acupuncture treatment for nocturnal enuresis, were considered. The primary outcome measure was change in the mean number of wet episodes following treatment. Electronic searching was supplemented by hand searching of western medicine and traditional Chinese medicine (TCM) journals along with English language alternative medicine journals. Trials were assessed for quality and sources of bias. Meta-analysis was performed and the overall weighted odds ratio (OR) and associated 95% confidence interval (CI) were computed using the fixed effect model; the Forest plot was used to demonstrate results. RESULTS: Two hundred six abstracts were identified, of which 11 studies were eligible for data extraction. All the trials were of low methodological quality. There was some evidence that acupuncture is useful for nocturnal enuresis when used in conjunction with other treatment that may also include a different form of acupuncture (OR 3.98, CI: 2.2-7.2). When one form of acupuncture is compared with another there was marked heterogeneity, implying that some forms of acupuncture are effective. CONCLUSION: This review provides tentative evidence for the efficacy of acupuncture for the treatment of childhood nocturnal enuresis. Due to the low methodological quality of studies, evidence to identify, which parameters of acupuncture work best, is lacking. More rigorous trials are clearly warranted. PMID- 15791607 TI - Intermittent disconjugate gaze: a novel finding in nonalcoholic steatohepatitis and cryptogenic cirrhosis. PMID- 15791609 TI - Percutaneous ablation for hepatocellular carcinoma. PMID- 15791608 TI - Impact of hepatitis C on health related quality of life: a systematic review and quantitative assessment. AB - Hepatitis C virus (HCV) diminishes health related quality of life (HRQOL), and it is now common to measure HRQOL in clinical trials. We sought to summarize the HRQOL data in HCV, and to establish the minimally clinically important difference (MCID) in HRQOL scores in HCV. We performed a systematic review to identify relevant studies, and converted HRQOL data from each study into clinically interpretable statistics. An expert panel used a modified Delphi technique to estimate the MCID in HCV. We found that patients with HCV scored lower than controls across all scales of the SF-36. Patients achieving sustained virological response (SVR) scored higher across all scales versus patients without SVR, especially in the physical health domains. HRQOL differences did not correspond with differences in liver histology or ALT levels. Based upon the published data, the expert panel concluded that the SF-36 vitality scale was most relevant in patients with HCV, and generated a mean MCID of 4.2 points on this scale. In conclusion, patients with HCV have a clinically significant decrement in HRQOL versus controls, and physical HRQOL improves in patients achieving SVR but not in those without SVR. The data further suggest that traditional outcomes fail to capture the full spectrum of illness related to chronic HCV. A difference of 4.2 points on the SF-36 vitality scale can be used as an estimate of the MCID in HCV, and this value may be used as the basis for power calculations in clinical trials evaluating HRQOL. Supplementary material for this article can be found on the HEPATOLOGY website (http://www.interscience.wiley.com/jpages/0270 9139/suppmat/index.html). PMID- 15791611 TI - Persistence of antibody after hepatitis B vaccine and the question of boosters. PMID- 15791613 TI - Progression of liver fibrosis in women infected with hepatitis C: long-term benefit of estrogen exposure. PMID- 15791615 TI - Abnormal concentrations of esterified carnitine in bile: a feature of pediatric acute liver failure with poor prognosis. AB - The etiology of acute liver failure in children is unknown in a large number of cases. Defects in fatty acid oxidation have been shown to lead to severe liver injury. This retrospective analysis examined the bile acylcarnitine profiles of 27 children with acute liver failure who underwent liver transplantation or died. Results were compared with 758 postmortem samples from individuals without acute liver failure. Cumulative amounts of free carnitine, medium- or long-chain species in excess of the 95th percentile of the control group were considered abnormal. Fourteen samples had normal profiles. Three had markedly elevated concentrations of free carnitine, whereas ten showed elevations in medium- or long-chain species. The relative risk of death was 2.86 (95% confidence interval, 1.08-7.54, P = .01) in the 10 children with elevated concentrations of medium- or long-chain species compared with those with normal analyses. Overall, medium- and long-chain acylcarnitines were increased in those patients who died compared with survivors, (dead vs. alive; medium-chain, 187 +/- 74 vs. 32 +/- 12 micromol/L, P = .008; long-chain, 146 +/- 74 vs. 15 +/- 8 micromol/L, mean +/- standard error of the mean, P = .018). These studies describe biliary free and esterified carnitine profiles in children with acute liver failure. In conclusion, the findings raise the hypothesis that abnormalities in fatty acid oxidation may predispose to a worse outcome in acute liver failure. PMID- 15791616 TI - Is HFE involved in increased hepcidin expression and hypoferremia in inflammation and anemia of chronic disease? PMID- 15791617 TI - Regulatory T cells contribute to the impaired immune response in patients with chronic hepatitis B virus infection. AB - Chronic hepatitis B virus (HBV) infection is characterized by a weak immune response to HBV. Regulatory T cells (T(reg)) can suppress the function of effector T cells and may thus be key players in this impaired immune response. Changes in the functionality or number of T(reg) could explain the decreased antiviral response in chronic HBV patients. To investigate the role of T(reg) in chronic HBV infection, we compared the proportional frequency and functionality of T(reg) in peripheral blood of 50 chronic HBV patients, 23 healthy controls, and 9 individuals with a resolved HBV infection. A higher percentage of T(reg), defined as CD4, CD25, CD45RO, and cytotoxic T-lymphocyte-associated antigen 4 positive cells, was detected within the population of CD4(+) cells in peripheral blood of chronic HBV patients compared with healthy controls and individuals with a resolved HBV infection. Accordingly, chronic HBV patients displayed a higher FoxP3 messenger RNA level than healthy controls. Depletion of CD25(+) cells from peripheral blood mononuclear cells (PBMC) of chronic HBV patients resulted in an enhanced proliferation after stimulation with HBV core antigen. Reconstitution of these depleted PBMC with CD4(+)CD25(+) T(reg) resulted in a dose-dependent reduction of both HBV-specific proliferation and interferon gamma production. In conclusion, chronic HBV patients harbor an increased percentage of T(reg) in peripheral blood compared with controls. T(reg) have an immunosuppressive effect on HBV-specific T helper cells. The presence of HBV-specific T(reg) could contribute to an inadequate immune response against the virus, leading to chronic infection. PMID- 15791618 TI - Two common PFIC2 mutations are associated with the impaired membrane trafficking of BSEP/ABCB11. AB - Progressive familial intrahepatic cholestasis type 2 (PFIC2) is caused by a mutation in the bile salt export pump (BSEP/ABCB11) gene. However, the mechanisms for the deficiency in the function of two mutations (E297G and D482G), which are frequently found in European patients, have not yet been identified. In the present study, we examined the transport activity and cellular localization of these two mutants in human embryonic kidney 293 and Madin-Darby canine kidney II cells, respectively. Introduction of E297G and D482G mutations into the human BSEP gene by site-directed mutagenesis resulted in a significant reduction in the BSEP expression level, which was associated with impaired membrane trafficking. Most of the D482G BSEP and some of the E297G BSEP underwent only core glycosylation and appeared to be predominantly located in the endoplasmic reticulum. The inhibition of proteasome function by MG132 resulted in the cellular accumulation of the core glycosylation form of the two mutants. In contrast, transport studies for taurocholate and glycocholate with membrane vesicles isolated from complementary DNA-transfected cells indicated that both mutations did not significantly affect the transport function of BSEP per se. In conclusion, E297G and D482G mutations result in impaired membrane trafficking, whereas the transport functions of these mutants remain largely unchanged. PMID- 15791619 TI - Leptin reverses nonalcoholic steatohepatitis in patients with severe lipodystrophy. AB - Severe lipodystrophy is characterized by diminished adipose tissue and hypoleptinemia, leading to ectopic triglyceride accumulation. In the liver, this is associated with steatosis, potentially leading to nonalcoholic steatohepatitis (NASH). We investigated the prevalence of NASH and the effect of leptin replacement in these patients. Ten patients with either generalized lipodystrophy (8 patients) or Dunnigan's partial lipodystrophy (2 patients) were included in this analysis. Paired liver biopsy specimens were obtained at baseline and after treatment with recombinant methionyl human leptin (r-metHuLeptin), mean duration 6.6 months. The extents of portal and parenchymal inflammation, steatosis, ballooning, presence of Mallory bodies, and fibrosis in liver biopsy specimens were scored using a previously validated system developed to assess NASH activity. Histological disease activity was defined as the sum of ballooning, steatosis, and parenchymal inflammation scores. We concurrently tested serum triglycerides and aminotransferases and estimations of liver volume and fat content by magnetic resonance imaging. Eight of 10 patients met histological criteria for NASH at baseline. After treatment with r-metHuLeptin, repeat histological examinations showed significant improvements in steatosis (P = .006) and ballooning injury (P = .005), with a reduction of mean NASH activity by 60% (P = .002). Fibrosis was unchanged. Significant reductions were seen in mean serum triglycerides (1206-->226 mg/dL, P = .002), glucose (220-->144 mg/dL, P = .02), insulin (46.4-->24.8 muIU/mL, P = .004), ALT (54-->24 U/L, P = .02), AST (47-->22 U/L, P = .046), liver volume (3209-->2391 cm(3), P = .007), and liver fat content (31-->11%, P = .006). In conclusion, r-metHuLeptin therapy significantly reduced triglycerides, transaminases, hepatomegaly, and liver fat content. These reductions were associated with significant reductions in steatosis and the hepatocellular ballooning injury seen in NASH. PMID- 15791620 TI - Compromised lymphocytes infiltrate hepatocellular carcinoma: the role of T regulatory cells. AB - Hepatocellular carcinoma (HCC) has a poor prognosis with limited therapeutic options. We propose that local immune responses in patients with HCC are held in check by tumor-infiltrating CD4(+)CD25(+) T-regulatory lymphocytes (T(reg) cells), which suppress the activity and proliferation of effector CD4(+) and CD8(+) T cells. The phenotype and cell cycle status of tumor-infiltrating lymphocytes (TILs) in HCC were analyzed via immunohistochemistry of sections from patients undergoing surgery for HCC and via flow cytometry of peripheral blood mononuclear cells and TILs isolated from patients with HCC. Circulating and tumor infiltrating T-cell function and activation status were assessed via proliferation and flow cytometry. More than 96% of TILs were quiescent as measured via Mcm-2 or Ki-67 expression, while less than 10% of CD8(+) T cells expressed perforin or granzyme B. CD4(+)CD25(+) T(reg) cells comprised 8.7% (1.4 13.8) of TILs and always exceeded the proportion in distant nontumor tissue (2.4% [1.5-5.6]; P = .014). T(reg) cells isolated from HCC suppressed proliferation of autologous circulating CD4(+)CD25(-) cells and perforin expression and proliferation of autologous CD8(+) T cells. The proportion of circulating T(reg) cells in patients with HCC was similar in healthy controls (7.2% [1.2-23.3] and 9.2% [1.6-30.2], respectively), but the proportion of circulating T(reg) cells that were also transforming growth factor beta1(+) was elevated in HCC compared with controls (55.5% [8.2-73.9] and 2.0% [0-4.9], respectively; P = .003). In conclusion, TILs are compromised and contain a subpopulation of suppressive CD4(+)CD25(+)Foxp3(+) T(reg) cells. Functional deletion of tumor-infiltrating T(reg) cells could enhance tumor-specific immunotherapy. PMID- 15791621 TI - The clinical development of percutaneous heart valve technology: a position statement of the Society of Thoracic Surgeons (STS), the American Association for Thoracic Surgery (AATS), and the Society of Cardiovascular Angiography and Intervention (SCAI). PMID- 15791622 TI - Mycotic aneurysm of left anterior descending artery after sirolimus-eluting stent implantation: a case report. AB - We report a case of mycotic aneurysms due to Staphylococcus aureus infection in the left anterior descending coronary artery in a 56-year-old male after implantation of a sirolimus-eluting stent. This is an unreported complication of a drug-eluting stent. PMID- 15791623 TI - Mild renal pyelectasis in the second trimester: determination of cut-off levels for postnatal referral. AB - OBJECTIVE: To establish guidelines for postnatal referral of fetuses presenting with mild pyelectasis in the second trimester of pregnancy. METHODS: In a retrospective study, 87 fetuses with a renal pelvis anteroposterior (RPAP) diameter of > or = 4 mm and < or = 10 mm before 28 weeks of gestation were included. All patients had a third-trimester scan and fetuses with an RPAP diameter of > or = 10 mm at that stage were referred for postnatal assessment. The family practitioner of all infants with an RPAP of < 10 mm in the third trimester was contacted for follow-up information. The RPAP diameter most predictive of renal pathology was determined with receiver-operating characteristics (ROC) curve analysis for both the first and second scans. RESULTS: In 36 of 87 infants, 49 abnormal kidneys were diagnosed. Seven infants required surgery on eight renal tracts. The ROC curves of the first scan, second scan and differences between scans resulted in an area under the curve of 0.60, 0.87 and 0.85, respectively. The sensitivities and specificities for a cut-off level of 8, 9 and 10 mm at the second scan were 80%, 71% and 61% and 79%, 90% and 93%, respectively. At a cut-off level of 10 mm, only cases of insignificant minimal dilatation and a case of vesicoureteric reflux (VUR) requiring surgery were not detected. CONCLUSION: After establishing a diagnosis of mild pyelectasis before 28 weeks, a second scan is mandatory to determine which infants need postnatal evaluation. A cut-off level of 8 mm has a low specificity but includes most cases of pathology. A cut-off level of 10 mm detects most significant pathology; however, VUR may not be detected. PMID- 15791624 TI - Bilateral lesion in the lateral columns and complete urinary retention: association with the spinal cord descending pathway for micturition. PMID- 15791626 TI - Screening test model using duration of labor for the detection of postpartum urinary retention. AB - AIMS: To construct a screening test model for postpartum urinary retention (PUR), using the duration of labor, which was shown to be the risk factor for PUR. METHODS: A group of 691 consecutive patients were recruited at postpartum day 1. Of the 691 patients, 101 (14.6%) had PUR. The 691 patients were computer randomized into two groups, 'A' and 'B.' A receiver operating characteristic (ROC) curve was constructed for Group A to determine the optimum cutoff value for screening PUR using the duration of labor. The cutoff value was then applied to Group B to determine the screening test characteristics of the duration of labor: sensitivity, specificity, predictive values, and likelihood ratios. RESULTS: An ROC curve with an area under curve (AUC) of 0.63 (95% CI 0.57-0.69, P < 0.0001) was constructed. A range of cutoff values (with specificity ranging from 0.60 to 0.99) for the duration of labor was determined and the test characteristics computed. According to the test characteristics the optimal cutoff value for the duration of labor was 700 min. This cutoff value for the duration of labor has a specificity of 0.95, negative predicative value of 0.86, and likelihood ratio for a positive test of 0.88. CONCLUSION: A screening test model using the duration of labor can be constructed to predict PUR. PMID- 15791625 TI - Morphological and biochemical characterization of a human liver in a uPA-SCID mouse chimera. AB - A small animal model harboring a functional human liver cell xenograft would be a useful tool to study human liver cell biology, drug metabolism, and infections with hepatotropic viruses. Here we describe the repopulation, organization, and function of human hepatocytes in a mouse recipient and the infections with hepatitis B virus (HBV) and hepatitis C virus (HCV) of the transplanted cells. Homozygous urokinase plasminogen activator (uPA)-SCID mice underwent transplantation with primary human hepatocytes, and at different times animals were bled and sacrificed to analyze plasma and liver tissue, respectively. The plasma of mice that were successfully transplanted contained albumin and an additional 21 human proteins. Liver histology showed progressive and massive replacement of diseased mouse tissue by human hepatocytes. These cells were accumulating glycogen but appeared otherwise normal and showed no signs of damage or death. They formed functional bile canaliculi that connected to mouse canaliculi. Besides mature hepatocytes, human hepatic progenitor cells that were differentiating into mature hepatocytes could be identified within liver parenchyma. Infection of chimeric mice with HBV or HCV resulted in an active infection that did not alter the liver function and architecture. Electron microscopy showed the presence of viral and subviral structures in HBV infected hepatocytes. In conclusion, human hepatocytes repopulate the uPA(+/+)-SCID mouse liver in a very organized fashion with preservation of normal cell function. The presence of human hepatic progenitor cells in these chimeric animals necessitates a critical review of the observations and conclusions made in experiments with isolated "mature" hepatocytes. Supplementary material for this article can be found on the HEPATOLOGY website (http://www.interscience.wiley.com/jpages/0270 9139/suppmat/index.html). PMID- 15791627 TI - Detrusor mitochondrial lipid peroxidation and superoxide dismutase activity in partial bladder outlet obstruction of rabbits. AB - OBJECTIVES: Bladder outlet obstruction has shown to damage detrusor mitochondria with impaired detrusor contractility. One likely cause for mitochondrial injury is reactive oxygen species (ROS)-induced damages, including lipid peroxidation injury. We designed this study to examine this hypothesis. MATERIALS AND METHODS: Placing a silicon ring around the bladder neck of male New Zealand rabbits induced bladder outlet obstruction. The bladders were removed 3 (N = 6), 7 (N = 6), and 14 days (N = 8) later. Sham operated animals (N = 6 for each time period) served as the controls. Contractile function of the bladder was assessed by the response of the detrusor strips to bethanechol and field stimulation. Detrusor mitochondrial superoxide dismutase (SOD) activity and mitochondrial content of malondialdehyde (MDA) were determined. Detrusor contents of phosphocreatine and adenine triphosphate (ATP) were assayed. RESULTS: (1) Outlet obstruction induced an increase in the bladder weight and a decrease in the contractile function; (2) mitochondrial SOD activity significantly elevated in every time period of the obstruction, indicating a persistently increased ROS generation; (3) detrusor MDA level increased in 3-day obstruction animals. It returned to the control level in 7- and 14-day groups; (4) phosphocreatine content was significantly reduced in every time period of the obstruction; (5) ATP content was significantly decreased in 3- and 7-day groups; while 14-day obstruction group contained similar level as the sham-operated group. CONCLUSIONS: This study shows that bladder outlet obstruction increases generation of ROS and enhances lipid peroxidation of detrusor mitochondria. The resulted mitochondrial damages might sustain, leading to persistently depressed energy production and impaired detrusor contractility. PMID- 15791628 TI - Correlation of abdominal leak point pressure with objective incontinence severity in men with post-radical prostatectomy stress incontinence. AB - AIMS: To determine if the abdominal leak point pressure (ALPP) correlates with objective incontinence severity in patients suffering from post-prostatectomy stress incontinence. METHODS: Twenty-nine men were evaluated for urinary incontinence after radical prostatectomy with videourodynamics and a 24-hr pad test. ALPP was determined with and without a 7-French urodynamics catheter and the lowest value was accepted. Six patients with urgency incontinence associated with detrusor overactivity or decreased bladder compliance were excluded leaving 23 patients for analysis. The relationship between the variables of ALPP, 24-hr pad weight, age and time from prostatectomy were examined with Pearson correlation. RESULTS: The mean age was 66.4 years (SD +/- 7.9, range: 45-81) and the median time from radical prostatectomy was 23 months (IQR = 14-64, range: 9 204). The mean ALPP was 92.8 cm H(2)O (SD +/- 42.4 cm H(2)O) and the mean pad weight was 279.1 g (SD +/- 238.3 g). There was only a weak inverse correlation between ALPP and 24-hr pad weight which was not statistically significant (r = - 0.191, P = 0.38). Age and time from prostatectomy did not significantly correlate with ALPP (r = - 0.122, P = 0.58 and r = - 0.23, P = 0.29, respectively). CONCLUSIONS: ALPP did not correlate significantly with the 24-hr pad test in patients with post-prostatectomy stress incontinence. This suggests that in this patient subset, the ALPP is a relatively poor predictor of incontinence severity and, therefore, has limited clinical value in the urodynamic evaluation of post prostatectomy incontinence. The urodynamic assessment of these patients should focus on the presence or absence of stress incontinence and on the presence of associated bladder dysfunction. PMID- 15791629 TI - Novel method of generating prostate-specific Cre-LoxP gene switching via intraductal delivery of adenovirus. AB - BACKGROUND: In order to facilitate elucidation of oncogene or tumor suppressor gene function on initiation and progression of prostate cancer, it would be advantageous to develop an effective method to generate spatially and temporally controlled gene modification in murine prostates. METHODS: Adenovirus expressing Cre-recombinase (Adeno-Cre) was intraductally injected into the prostate of ROSA26 reporter mice. Immmunohistochemical and X-gal staining were performed on prostate tissue sections harvested from mice at various time points following viral injection to confirm expression and activity of Cre-recombinase, respectively. RESULTS: Adenovirus was intraductally delivered to the anterior lobe of the mouse prostate. Using this method of intraductal injection, we were able to precisely obtain Adeno-Cre infection to a majority of epithelial cells but not in the stromal cells or other organs. We further demonstrated that Adeno Cre infected epithelial cells not only expressed Cre-recombinase enzyme but more importantly, Cre-recombinase activity was revealed through positive X-gal staining in Rosa26 reporter mice, thus, confirming epithelial-specific Cre-loxP recombination in Adeno-Cre infected prostate tissue sections. CONCLUSIONS: This novel method of direct genetic delivery into adult murine prostates could provide an alternative to the more expensive and time-consuming transgenic/knockout approaches. The latter also have other limitations such as the availability of cell-type specific or temporally-regulated promoters, and the complication of genetic background differences, which can potentially be complemented by the technology we describe here. PMID- 15791630 TI - Contractile properties of the developing fetal sheep bladder. AB - AIM: To characterise the developmental changes to the normal bladder by examining the in vitro contractile properties of the fetal sheep detrusor smooth muscle bladder at different gestational ages. METHODS: Three groups of normally developing fetal sheep bladders were investigated: (1) 65-70 days gestation (2nd trimester); (2) 105-110 days (3rd trimester); (3) 135-140 days (term). Isometric contractions in isolated detrusor strips were measured during either electrical field stimulation (EFS) or exposure to agonists: carbachol, alpha-beta methylene ATP (ABMA) and KCl. RESULTS: There was a significant increase of absolute force elicited by EFS with fetal age. Contractures elicited by carbachol, ABMA and KCl increased between Groups 1 and 2, but not thereafter. The proportional increase of contractions elicited by carbachol and ABMA was also greater between Groups 1 and 2, than for EFS and KCl. CONCLUSIONS: Fetal development between 65 and 140 days in the sheep is associated with increased contractile activation. The data are consistent with an increase of muscle development in the earlier stages (65 110 days). In the later stage (110-140 days) muscle development is complete but functional innervation of the tissue continues. PMID- 15791631 TI - Rho-kinase inhibition suppresses bladder hyperactivity in spontaneously hypertensive rats. AB - AIMS: Spontaneously hypertensive rats (SHR) exhibit overactive bladder (OAB) symptoms and have an up-regulated calcium sensitizing RhoA/Rho-kinase pathway in their vascular smooth muscle tissues. This study examined the role of RhoA/Rho kinase pathway in bladder hyperactivity by evaluating the effect of a specific Rho-kinase inhibitor (Y-27632) on SHR bladder function. METHODS: Adult male SHR (n = 9) and their normotensive controls (Wistar-Kyoto; WKY) (n = 8) were anesthetized and the carotid artery cannulated for blood pressure monitoring. A catheter was fixed into the bladder dome and connected to a pressure transducer and an infusion pump. After equilibration, systemic and bladder pressure were recorded. Continuous filling cystometrograms (CMGs) were performed and threshold pressure (TP), peak pressure (PP), and number of voids and non-voiding contractions (NVCs) per unit time recorded. Each SHR then received Y-27632, 10 mumol intra-arterially. After 10 min, CMG was repeated and the same measurements recorded. Bladder tissues were evaluated immunohistochemically (IHC) for RhoA protein expression. RESULTS: SHR exhibited significantly higher number of voids and NVCs than normotensive WKY rats (P < 0.05). In SHR, Y-27362 administration significantly decreased the number of voids (29%, from 0.83 +/- 0.3 to 0.63 +/- 0.17 voids/min) and NVCs (61%, from 1.8 +/- 0.54 to 0.64 +/- 0.167 NVC/min). IHC showed significantly higher RhoA protein expression in SHR bladder tissues. CONCLUSIONS: Overexpression of RhoA may play a role in hypertension-related OAB. Inhibition of Rho-kinase activity with Y-27632 produced a significant suppression of bladder overactivity. Identification of Rho-kinase isoforms that are bladder tissue specific and their selective inhibitors may help to disassociate the unwanted hypotensive effects of this approach. PMID- 15791632 TI - Urgency of micturition and detrusor contractility in men with prostatic obstruction and overactive bladders. AB - AIMS: In men with prostatic obstruction and detrusor overactivity (DO), to ascertain whether urgency of micturition affects bladder contractility. MATERIALS AND METHODS: We urodynamically assessed five groups of 20 men each who had bladder outflow obstruction (BOO) from benign prostatic enlargement-Groups 1 (with no DO and no urgency), 2 (with DO and no urgency), 3A (with DO and moderate urgency), 3B (with DO and severe urgency), and 4 (with DO, severe urgency and chronic ischemic cerebral lesions). Urgency was graded as moderate or severe by the ability to avert an urgent void at cystometry for > or =2 or <2 min, respectively. BOO was assessed by the "Abrams-Griffiths number" (AG) and bladder contractility by the parameters PIP and WF(max). RESULTS: AG did not differ significantly in Groups 2, 3A, and 3B, proved higher in such groups than in Group 1, and was nearly the same in Groups 1 and 4. PIP and WF(max) were significantly higher in Groups 2, 3A, and 3B than in Groups 1 and 4, had the highest levels in Group 3B, and did not differ significantly in Groups 1-4 and 2-3A. CONCLUSIONS: In DO patients with prostatic obstruction there seems to be a DO-related facilitation of bladder contractility. In the same patients, severe urgency of micturition might over-amplify (i.e., enhance a DO-related facilitation of) bladder contractility, provided there are no neurogenic (chronic ischemic cerebral) lesions. PMID- 15791634 TI - Effects of serotonin 5-HT1A agonist in advanced Parkinson's disease. AB - Intermittent stimulation of striatal dopaminergic receptors seems to contribute to motor dysfunction in advanced Parkinson's disease (PD). With severe dopaminergic denervation, exogenous levodopa is largely decarboxylated to dopamine in serotonergic terminals. If 5-HT1A autoreceptors regulate dopamine as well as serotonin release, in parkinsonian patients inhibition of striatal serotonergic neuron firing might help maintain more physiological intrasynaptic dopamine concentrations and thus ameliorate motor fluctuations and dyskinesias. To evaluate this hypothesis, effects of a selective 5-HT1A agonist, sarizotan, given orally at 2 and 5 mg twice daily to 18 relatively advanced parkinsonian patients, were compared with baseline placebo function during a 3-week, double blind, placebo-controlled, proof-of-concept study. Sarizotan alone or with intravenous levodopa had no effect on parkinsonian severity. But at safe and tolerable doses, sarizotan coadministration reduced levodopa-induced dyskinesias and prolonged its antiparkinsonian response (P < or = 0.05). Under the conditions of this study, our findings suggest that 5-HT1A receptor stimulation in levodopa treated parkinsonian patients can modulate striatal dopaminergic function and that 5-HT1A agonists may be useful as levodopa adjuvants in the treatment of PD. PMID- 15791633 TI - Pelvic floor muscle training is effective in women with urinary incontinence after stroke: a randomised, controlled and blinded study. AB - AIMS: The aim of this study was to evaluate the effect of Pelvic Floor Muscle Training (PFMT) in women with urinary incontinence (UI) after ischemic stroke. MATERIALS AND METHODS: Three hundred and thirty-nine medical records of stroke patients were searched. Twenty-six subjects were randomised to a Treatment Group (14 subjects) or a Control Group (12 subjects) in a single blinded, randomised study. The intervention included 12 weeks of standardised PFMT. The outcome measures were: (1) diary recording the frequency of voiding, the number of incontinence episodes and used pads; (2) 24-hr home pad test; and (3) vaginal palpation of pelvic floor muscle evaluating function, strength, static and dynamic endurance. RESULTS: Twenty-four subjects with urge, stress and mixed stress/urge incontinence, completed the study, 12 in each group. A significant improvement in frequency of voiding in daytime (Treatment Group/Control Group: 7/8 at pre-test, 6/9 at post-test (median values), P=0.018), 24-hr pad test (Treatment Group/Control Group: 8/12 to 2/8 g P=0.013) and dynamic endurance of pelvic floor muscle (Treatment Group/Control Group: 11/20 to 20/8 contractions of Pelvic Floor Muscle, P=0.028) was demonstrated in the Treatment Group compared to the Control Group. A significant improvement in frequency of voiding in daytime (decreased from seven to six, P=0.036), pelvic floor muscle function (P=0.034), strength (P=0.046), static endurance increased from 9 to 30 sec (P=0.028) and dynamic endurance increased from 11 to 20 contractions (P=0.020) was also demonstrated within the Treatment Group, but not in the Control Group. CONCLUSION: PFMT had a significant effect in women with UI after stroke measured by diaries, pad tests and vaginal palpation. PMID- 15791635 TI - Factors influencing self-perceived disease severity in women with stress urinary incontinence combined with or without urge incontinence. AB - AIMS: The aim of this study was to assess the impact of patient-perceived disease severity (PPDS) on the quality of life (QoL) of women with urinary incontinence (UI) and to identify factors predicting PPDS. METHODS: A total of 109 women (mean age 54.9; range 31-77) with stress UI combined with or without urge UI were included in the primary analyses. The incontinence quality of life (I-QoL) devised during the course of this study was used to assess the QOL impact of UI. RESULTS: PPDS of women with UI increased as I-QoL scores decreased (P<0.001). When analyzed by patient characteristics and objective test results, PPDS increased only with the number of episodes (P=0.005) and pad test weight increased (P=0.010). By multivariate regression analysis, patients who complained of UI "three to four times a day or more" had 6.4-fold higher risk (P=0.027) of perceiving that their symptoms were more severe than those who complained of a UI "one to two times per week or less." Patients with a pad test weight of >25 g had a 4.7-fold higher risk of perceiving their symptoms were more severe than those with a pad test weight of <15 g. CONCLUSIONS: Our results suggest that the frequency of UI episodes and the volume of urine loss are associated with PPDS. In addition, the I-QoL scores deteriorated significantly as the PPDS of incontinence increased. Thus, PPDS may impact on the QoL of women with stress UI combined with or without urge UI. PMID- 15791636 TI - Cytomics goes 3D: toward tissomics. PMID- 15791637 TI - Homoserine-derived cyclic sulfamidate as chiral educt for the diversity-oriented synthesis of lactam-bridged dipeptides. AB - Introduction of structural constraint into peptides is an effective way for studying their conformation-activity relationships. Conformationally restrained dipeptidyl lactams, important building blocks for the synthesis of peptidomimetics, have now been synthesized from N-[9-(9-phenylfluorenyl)]-L aspartic acid alpha-cumyl beta-methyl diester as an inexpensive chiral educt. After selective reduction of the beta-methyl ester with diisobutylaluminum hydride (DIBAL-H), homoserine was treated with thionyl chloride, imidazole, and triethylamine to give sulfamidites. Diastereoisomers were separated by chromatography and oxidation of the major sulfamidite (2R,4S)- with catalytic ruthenium trichloride afforded sulfamidate. A series of gamma-lactam-bridged dipeptides was then obtained by ring opening of sulfamidate cumyl ester with a series of amino esters, selective cumyl ester removal, and lactam formation. The resulting dipeptidyl lactams possessed aliphatic, aromatic, amino, thioether, and carboxylate side chains. A gamma-lactam analog of Pro-Leu-Gly-NH2 (PLG), was synthesized to illustrate the potential for using this approach in the synthesis of biologically active peptide mimics. PMID- 15791640 TI - LIM-homeodomain genes as territory markers in the brainstem of adult and developing Xenopus laevis. AB - We investigated expression patterns of the LIM-homeodomain (LIM-hd) genes x-Lhx1, x-Lhx2, x-Lhx5, and x-Lhx9 in the brainstem of Xenopus laevis during larval development and in the adult. The two groups of paralogous genes, x-Lhx1/x-Lhx5 and x-Lhx2/x-Lhx9, showed fundamentally different expression patterns, being expressed in ventral versus dorsal territories of the midbrain and hindbrain, respectively. Indeed, prominent expression of x-Lhx1/5 was found in the mesencephalic tegmentum and the hindbrain reticular formation, whereas conspicuous x-Lhx2/9 expression was observed in the torus semicircularis and isthmic nucleus. A few shared expression domains for the two pairs of paralogs included the optic tectum and the anterodorsal and pedunculopontine nuclei. In each structure, expression of the two paralogs was almost identical, indicating that the regulation of their expression in this part of the brain has evolved slightly since gene duplication occurred. Notable exceptions included the expression of x-Lhx1 but not x-Lhx5 in the Purkinje cells and the expression of x Lhx9 but not x-Lhx2 in the lateral line nucleus. The analysis of LIM-hd expression patterns throughout development allowed the origin of given structures in early embryos to be traced back. x-Lhx1 and x-Lhx5 were relevant to locate the cerebellar anlage and to follow morphogenesis of the cerebellar plate and cerebellar nuclei. They also highlighted the rhombomeric organization of the hindbrain. On the other hand, x-Lhx2 and x-Lhx9 showed a dynamic spatiotemporal pattern relative to tectal development and layering, and x-Lhx9 was useful to trace back the origin of the isthmus in early development. PMID- 15791641 TI - Expression of soluble guanylyl cyclase in rat cerebral cortex during postnatal development. AB - Soluble guanylyl cyclase (sGC), the principle "receptor" for nitric oxide (NO), catalyzes the formation of cyclic guanosine monophosphate (cGMP), an intracellular second messenger. Studies in invertebrates have shown that the NO/cGMP pathway is involved in several aspects of neural development, including neuronal migration, dendritic and axonal outgrowth, and synaptogenesis. In vitro studies suggest a developmental role also in mammals. To investigate whether the NO/cGMP pathway might mediate these processes in vivo, we performed immunohistochemistry for sGC on sections from postnatal rat cerebral cortex. Early in postnatal development, migrating neurons in the cortical plate were immunonegative, whereas neurons deeper in the cortex that had completed migration were immunopositive. At the subcellular level, sGC preferentially stained dendrites rather than axons, but, at postnatal day 1 (PND1), sGC was found in a large fraction of axonal growth cones, especially those oriented toward the pial surface. At PND10-20 (the period of maximal synaptogenesis), sGC immunostaining was located mainly in dendritic shafts and was only occasionally associated with spines or axon terminals. These results support a role for the NO/cGMP pathway in dendritic development but argue against a major role in neuronal migration and synaptogenesis. PMID- 15791642 TI - Pathway sampling by regenerating peripheral axons. AB - A century ago, Ramon y Cajal described the generalized response of regenerating peripheral axons to their environment. By using mice that express fluorescent proteins in their axons, we are now able to quantify the response of individual axons to nerve transection and repair. Sciatic nerves from nonexpressing mice were grafted into those expressing a yellow variant of green fluorescent protein, then examined at 5, 7, or 10 days after repair. Regeneration was found to be a staggered process, with only 25% of axons crossing the repair in the first week. In the setting of Wallerian degeneration, this stagger will expose growth cones to an evolving menu of molecular cues upon which to base pathway decisions. Many axons arborize, allowing them to interact simultaneously with several pathways. Arborization could serve as the anatomical substrate for specificity generation through collateral pruning. Axons often travel laterally across the face of the distal stump before choosing a pathway. As a result, the average unbranched axon has access to over 100 distal Schwann cell tubes. This extensive access, however, does not ensure correct matching of axon and end organ, suggesting that pathway choice is made on the basis of factors other than end organ identity. These observations explain the failure of refined surgical techniques to restore normal function after nerve injury. The apparent wandering of axons across the repair defies surgical control and mandates a biological approach to reuniting severed axons with appropriate distal pathways. PMID- 15791643 TI - Development of the projection from the nucleus of the brachium of the inferior colliculus to the superior colliculus in the ferret. AB - Neurons in the deeper layers of the superior colliculus (SC) have spatially tuned receptive fields that are arranged to form a map of auditory space. The spatial tuning of these neurons emerges gradually in an experience-dependent manner after the onset of hearing, but the relative contributions of peripheral and central factors in this process of maturation are unknown. We have studied the postnatal development of the projection to the ferret SC from the nucleus of the brachium of the inferior colliculus (nBIC), its main source of auditory input, to determine whether the emergence of auditory map topography can be attributed to anatomical rewiring of this projection. The pattern of retrograde labeling produced by injections of fluorescent microspheres in the SC on postnatal day (P) 0 and just after the age of hearing onset (P29), showed that the nBIC-SC projection is topographically organized in the rostrocaudal axis, along which sound azimuth is represented, from birth. Injections of biotinylated dextran amine-fluorescein into the nBIC at different ages (P30, 60, and 90) labeled axons with numerous terminals and en passant boutons throughout the deeper layers of the SC. This labeling covered the entire mediolateral extent of the SC, but, in keeping with the pattern of retrograde labeling following microsphere injections in the SC, was more restricted rostrocaudally. No systematic changes were observed with age. The stability of the nBIC-SC projection over this period suggests that developmental changes in auditory spatial tuning involve other processes, rather than a gross refinement of the projection from the nBIC. PMID- 15791644 TI - Connexin36 mediates gap junctional coupling of alpha-ganglion cells in mouse retina. AB - Alpha-ganglion cells are present in all vertebrate retinae and are subdivided into ON and OFF types according to their level of dendritic ramification within the inner plexiform layer. They have large dendritic fields and usually a good responsiveness to moving stimuli. They were the first ganglion cells in which tracer coupling was observed, suggesting the presence of gap junctions composed of unknown connexins. Here we show that ON-alpha-ganglion cells in the mouse retina are coupled to amacrine cells, whereas OFF-alpha-ganglion cells are coupled to other OFF-alpha-ganglion cells and to amacrine cells. These tracer coupling patterns were completely absent in mice deficient in connexin36 (Cx36). The expression of Cx36 protein in alpha-ganglion cells but not in coupled amacrine cells was confirmed in mice in which the Cx36 coding DNA was replaced by the lacZ reporter gene. The dendritic localization and the distribution pattern of Cx36 patches, analyzed in mice in which the enhanced green fluorescent protein (EGFP) was linked to the C-terminal region of the Cx36 protein, revealed a rather small number of fluorescent plaques and different patterns for ON- and OFF-alpha ganglion cells. Furthermore, tracer coupling between OFF-alpha-ganglion cells could be inhibited by quinine, a gap junctional blocker with a slight preference for gap junctions formed by Cx36. These data strongly suggest that Cx36 gap junction channels are functional not only in interneurons but also in output neurons of the retina and are responsible for distinct coupling patterns of ganglion cells. PMID- 15791646 TI - Short-term physiologic effects of mechanical flow sorting and the Becton Dickinson cell concentrator in cultures of the marine phytoflagellata Emiliania huxleyi and Micromonas pusilla. AB - BACKGROUND: In contrast to large, high-efficiency cytometers, mechanically sorting benchtop instruments provide a feasible alternative for shipboard cell sorting of oceanic microbial communities. However, sorting efficiency of these instruments is constrained by their maximum sorting rate of approximately 300 cells/s and by constant dilution of sorted samples by sheath flow. These factors often render too low sorted cell concentrations for postsorting experiments of oceanic phytoplankton populations of low natural abundance. A Cell Concentrator module has been marketed to overcome these dilution effects. Postsorting experiments also have to consider potential physiologic effects of cell sorting. Short-term physiologic effects on phytoplankton photosynthetic rates and esterase activities by mechanical flow sorting and cell concentration and on the efficiency of the Cell Concentrator module are evaluated. METHODS: Increasing numbers of the oceanic phytoflagellates Micromonas pusilla and Emiliania huxleyi were sorted and concentrated, and recovery in the concentrated samples was compared with the sorted-only samples (concentration rate) and the total number of sorted cells (recovery rate). Photosynthetic rates and metabolic activities of sorted and sorted/concentrated cells were compared with unsorted cells. Photosynthetic rates were estimated from 14CO2 uptake experiments and metabolic activity quantified cytometrically after cleavage of fluorescein diacetate. RESULTS: Irrespective of the total number of sorted cells, concentration rates between concentrated and sorted cells remained mostly below 10-fold and did not increase with the number of concentrated cells. Recovery rates in the concentrated samples amounted to fewer than 10% of total sorted cells, except for forceful resuspension attempts in the Concentrator insert (25-44%), which might be unsuitable for delicate species. Cell sorting resulted in a 24-49% decrease in photosynthetic rates. Metabolic activity within metabolically active cells was not affected by cell sorting, but the share of metabolically active cells decreased by 32-37%. Cell concentration did not affect metabolic activity or the fraction of active cells but did increase photosynthetic rate several-fold compared with unsorted cells. CONCLUSION: Low recovery of concentrated cells, probably due to cell adhesion to the filer bottom of the Concentrator insert, render the Cell Concentrator of limited use to overcome dilution problems of mechanical flow sorting, particularly when results are extrapolated to natural, low-abundance populations. Severe changes in photosynthetic rates also render concentrated cells suspicious for subsequent physiologic experiments. Mechanical sorting alone also exhibited significant physiologic effects on sorted cells, some of which might not be temporary. Comparable effects between mechanical sorting and droplet sorting as previously reported confirm that physiologic effects might be caused predominantly by shear stress and laser exposure during cytometric analysis rather than the sorting process. Sufficient recovery time must be allowed before postsorting experiments, but potential changes in cell physiology from the natural conditions during postsorting recovery must be considered. PMID- 15791645 TI - Architecture and neurocytology of monkey cingulate gyrus. AB - Human functional imaging and neurocytology have produced important revisions to the organization of the cingulate gyrus and demonstrate four structure/function regions: anterior, midcingulate (MCC), posterior (PCC), and retrosplenial. This study evaluates the brain of a rhesus and 11 cynomolgus monkeys with Nissl staining and immunohistochemistry for neuron-specific nuclear binding protein, intermediate neurofilament proteins, and parvalbumin. The MCC region was identified along with its two subdivisions (a24' and p24'). The transition between areas 24 and 23 does not involve a simple increase in the number of neurons in layer IV but includes an increase in neuron density in layer Va of p24', a dysgranular layer IV in area 23d, granular area 23, with a neuron-dense layer Va and area 31. Each area on the dorsal bank of the cingulate gyrus has an extension around the fundus of the cingulate sulcus (f 24c, f 24c', f 24d, f 23c), whereas most cortex on the dorsal bank is composed of frontal motor areas. The PCC is composed of a dysgranular area 23d, area 23c in the caudal cingulate sulcus, a dorsal cingulate gyral area 23a/b, and a ventral area 23a/b. Finally, a dysgranular transition zone includes both area 23d and retrosplenial area 30. The distribution of areas was plotted onto flat maps to show the extent of each and their relationships to the vertical plane at the anterior commissure, corpus callosum, and cingulate sulcus. This major revision of the architectural organization of monkey cingulate cortex provides a new context for connection studies and for devising models of neuron diseases. PMID- 15791647 TI - Involvement of protein kinase PKN1 in G2/M delay caused by arsenite. AB - PKN1 is a serine/threonine protein kinase that has been reported to mediate cellular response to stress. We show here that in response to arsenite exposure, PKN1 kinase activity was stimulated, which was associated with increased binding of PKN1 to Cdc25C and delayed mitotic entry. A role for PKN1 in mediating arsenite-induced G(2)/M delay was supported by the finding that expression of a constitutively active form of PKN1 (PKN1AF3) in HeLa cells delayed the mitotic entry of cell cycle. Further experiments indicate that PKN1 directly phosphorylated serine 216 (Ser216) in Cdc25C, which then facilitated association between Cdc25C and 14-3-3. Significantly, expression of a phosphorylation mutant of Cdc25C (S216A) partially abrogated the cell-cycle arrest in response to arsenite. Together, our results suggest that PKN1 mediates arsenite-induced delay of the G(2)/M transition by binding to and phoshorylating Cdc25C. PMID- 15791648 TI - Functional consequences of mutations in a putative Akt phosphorylation motif of B raf in human cancers. AB - Mutations in the B-raf gene have been reported in a number of human cancers, including melanoma and lung cancer. More than 80% of the reported B-raf mutations were V599E; however, non-V599E mutations have been frequently found in non-small cell lung cancers as compared with melanoma. Some non-V599E mutations have been found surrounding Thr439, which is thought likely to be one of the three Akt phosphorylation sites in the B-raf protein. However, as a previous report indicated that Thr439 was not phosphorylated by Akt, the functional consequences of these mutations have been unclear. Here, we examined the effects of cancer related B-raf mutations surrounding Thr439 on the activation of the mitogen activated protein/ extracellular signal-regulated kinase kinase (MEK)/extracellular signal-regulated kinase (Erk) pathway and the transformation of NIH 3T3 fibroblasts. Among the three reported mutations (K438Q, K438T, and T439P) found in non-small cell lung carcinoma and melanoma, none elevated the activity of the MEK/Erk cascade as determined by in vitro kinase assays, immunoblots using antibody specific for phosphorylated Erk, or Elk1-dependent reporter assays. The inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt signaling by LY294002 increased the Erk activation induced by the mutant B-raf proteins, as well as by wild-type B-raf. Furthermore, the B-raf mutants did not have increased NIH 3T3-transforming activities, as determined by colony-formation assays. These results suggest that the B-raf mutations surrounding Thr439 found in human cancers are unlikely to contribute to increased oncogenic properties of B-raf. PMID- 15791650 TI - Dys-regulation of clusterin in human hepatoma is not associated with tumorigenesis but is secondary to cell response to external tresses. AB - It becomes feasible to perform genome-wide differential gene or protein expression in the post genome era. However, little has been addressed on the effects of external stresses and microenvironment alterations on the outcomes of gene and protein expression. To identify downregulated genes during hepatoma development, we combined the cDNA representational difference analysis (RDA) and reverse Northern blot analysis identifying eight genes. Of interest, the expression of the clusterin gene was either down or upregulated in 8 and 7 out of the 20 hepatoma tissues, respectively. Further analysis revealed that its expression was independent of patients' age, gender, causes of liver disease, tumor size, tumor histological stage, or clinical outcome, but was strongly associated with the methods of hepatectomy procedures. In vitro studies disclosed that the clusterin mRNA was increased twofold in early exponential phase of cell proliferation followed by downregulation in the subsequent quiescence phase, whereas it was rapidly increased up to twelvefold upon UV-induced apoptosis. These results suggest that dys-regulation of the clusterin gene in human hepatoma was most likely due to cellular responses to external stresses especially during the procedures for sample collection rather than any correlation to hepatoma development or progression. Our findings that external stresses or microenvironmental changes could greatly affect gene or protein expression offer a general caution to all the studies conducted via genomic and proteomic approaches. PMID- 15791649 TI - Differential inhibition of UVB-induced AP-1 and NF-kappaB transactivation by components of the jun bZIP domain. AB - Potential targets for chemoprevention of nonmelanoma skin cancer include UV induced nuclear factor kappaB (NF-kappaB) and activator protein-1 (AP-1) activation in keratinocytes. Inhibition of both these ultraviolet light B (UVB) induced transcription factors has been shown with the dominant-negative c-jun mutant, TAM67; however, its mechanism of action has not yet been determined. Here we demonstrated that transient transfection of a mouse keratinocyte cell line (308) with a dominant-negative phosphorylation mutant of c-Jun before exposure to 250 J/m(2) UVB inhibits transactivation mediated by both AP-1 and NF-kappaB transcription factors to levels below those of UVB exposed controls. Through the utilization of immunoprecipitation techniques, protein-protein interactions between NF-kappaB family members IkappaBalpha, IkappaBbeta, p50, and p65 (Rel-A) were identified with an Xpress tagged dominant-negative c-Jun (TAM67) protein. Expression of the leucine zipper domain of the TAM67 protein inhibited UVB induced NF-kappaB transactivation but not AP-1 transactivation. Expression of the bZIP domain of the TAM67 protein was able to inhibit transactivation mediated by both transcription factors. These data demonstrate that TAM67 is able to inhibit two significant UVB-induced molecular targets AP-1 and NF-kappaB, and that the inhibition of these two transcription factor families is potentially due to protein-protein interactions between different regions of the dominant-negative c Jun protein. PMID- 15791651 TI - Hibiscus polyphenol-rich extract induces apoptosis in human gastric carcinoma cells via p53 phosphorylation and p38 MAPK/FasL cascade pathway. AB - In view of the continuing need for effective anticancer agents, and the association of diet with reduced cancer risk, edible plants are increasingly being considered as sources of anticancer drugs. Hibiscus sabdariffa Linne (Malvaceae), an attractive plant believed to be native to Africa, is cultivated in the Sudan and Eastern Taiwan. Polyphenols had been demonstrated previously to possess antioxidative and antitumor promoting effects. In this study, investigations were conducted to examine the mechanism of the anticancer activity of H. sabdariffa L., Hibiscus polyphenol-rich extracts (HPE). Using HPLC assay, HPE was demonstrated to contain various polyphenols. HPE induced cell death of eight kinds of cell lines in a concentration-dependent manner. Among them human gastric carcinoma (AGS) cells were the most susceptible to HPE (0.95 mg/mL HPE inhibited its growth by 50%). Our results revealed that AGS cells underwent DNA fragmentation, and had an increase in the distribution of hypodiploid phase (apoptotic peak, 52.36%) after a 24-h treatment with HPE (2.0 mg/mL). This effect of HPE in AGS cells might be mediated via p53 signaling and p38 MAPK/FasL cascade pathway, as demonstrated by an increase in the phosphorylation of p53 and the usage of a specific p38 inhibitor, SB203580. Thus, our data present the first evidence of HPE as an apoptosis inducer in AGS cells and these findings may open interesting perspectives to the strategy in human gastric cancer treatment. PMID- 15791652 TI - Comparison of laser beam intensity profiles produced by photodynamic therapy (PDT) and transpupillary thermotherapy (TTT) lasers. AB - BACKGROUND AND OBJECTIVES: For photodynamic therapy (PDT) or transpupillary thermotherapy (TTT) lasers, long irradiation time (typically 1 minute or longer) is used and a large area of retina is treated. Consequently, the power stability but also the light distribution within the laser beam plays a major role. This study aimed to evaluate beam intensity profiles produced by several PDT and TTT lasers. STUDY DESIGN/MATERIALS AND METHODS: A beam profile analyzer (Cohu 4812 camera connected to a LPA-300PC, Spiricon, Logan, UT) was used to compare the beam profiles of PDT lasers: OPAL (Lumenis, USA); ACTIVIS (Quantel Medical, France), VISULAS (Zeiss, Germany). Spots of 2, 3, 4, and 5 mm were tested with each laser. Similarly, TTT lasers: OCULIGHT SLx (Iridex, CA) and IRIDIS trade mark (Quantel Medical, France) were evaluated with 2 and 3 mm spot diameter and power ranging from 200 to 1,000 mW. RESULTS: PDT lasers: OPAL had a "top hat" and homogeneous profile whatever the spot size. Numerous micro-spikes and micro nadirs of power were observed with the ACTIVIS and the VISULAS. TTT lasers: for the IRIDIS the beam shape was rather gaussian, but the homogeneity was reduced by micro-spikes of power. With the OCULIGHT Slx the beam shape was rather top hat and only few micro-spikes or micro-nadirs of power could be disclosed. DISCUSSION: The literature tends to prove that the shape and homogeneity of the beam profile could play a role on the efficacy of the treatment. CONCLUSION: Since PDT and TTT lasers display different beam profiles, this parameter should be carefully evaluated when performing clinical evaluations of PDT or TTT treatments. PMID- 15791653 TI - Morphological and 12S rRNA gene comparison of two Branchiostoma species in Xiamen waters. AB - Until now all amphioxus living in Xiamen waters have been regarded as Branchiostoma belcheri without any suspicion. However, a study based on Cyt b gene sequence comparisons of Branchiostoma belcheri (Oucuo in Xiamen waters) and Branchiostoma belcheri tsingtauense (Japanese waters) showed unexpected high divergence suggesting that the taxonomic status of Xiamen amphioxus should be reevaluated. In order to clarify this issue, we collected the animals from two sampling sites (Oucuo and Huangcuo in Xiamen waters), and compared their morphologies (meristic and non-meristic) as well as the complete sequences of their 12S rRNA genes. The samples could be distinguished by six of the non meristic traits and five of the meristic traits. Moreover, the genetic distance based on 12S rRNA gene between Oucuo and Huangcuo amphioxus is 21.13%, but that between Huangcuo and Japanese amphioxus is only 0.56%. Our results suggest that original subspecies B. belcheri tsingtauense should be elevated to species level, becoming B. tsingtauense. Therefore, two species of genus Branchiostoma are living in Xiamen waters. One is the original species B. belcheri (Oucuo) and the other is B. tsingtauense (Huangcuo). PMID- 15791654 TI - Unemployment, labor force composition and sickness absence: a panel data study. AB - Sickness absence tends to be negatively correlated with unemployment rates. In addition to pure health effects, this may be due to moral hazard behavior by workers who are fully insured against income loss during sickness and to physicians who meet demand for medical certificates. Alternatively, it may reflect changes in the composition of the labor force, with more sickness-prone workers entering the labor force in upturns. A panel of Norwegian register data is used to analyze long-term sickness absences. The unemployment rate is shown to be negatively associated with the probability of absence, and with the number of days of sick leave. Restricting the sample to workers who are present in the whole sample period, the negative relationship between absence and unemployment becomes clearer. This indicates that procyclical variations in sickness absence are caused by established workers and not by the composition of the labor force. PMID- 15791656 TI - Cysteinylation of maternal plasma albumin and its association with intrauterine growth restriction. AB - OBJECTIVES: We investigated cysteinylation of maternal plasma albumin in an observational study of intrauterine growth restriction (IUGR). High-risk pregnancies and uteroplacental insufficiency (UPI) have been associated with elevated levels of homocysteine, and, in oxidizing environments, homocysteine is converted to cysteine, resulting in cysteinylation of proteins. METHODS: The study population included pregnancies with IUGR (n = 12) and uncomplicated pregnancies as controls (n = 8). In all cases, cysteinylation of maternal plasma albumin was measured in a blinded fashion using liquid chromatography coupled with electrospray ionization mass spectrometry. RESULTS: Markedly elevated maternal plasma levels of cysteinylated albumin were detected in pregnancies with IUGR (44.7 +/- 14.8% of total albumin) compared to those in normal pregnancies (20.9 +/- 6.1%). As a result, native albumin decreased from 52.5 +/- 6.5% of total albumin in normal pregnancies to 30.1 +/- 13.3% in IUGR pregnancies. CONCLUSIONS: We hypothesize that sustained oxidative stress present in UPI is reflected by high levels of maternal cysteinylated albumin and may be a factor in the etiology of IUGR. Results of this preliminary study suggest that measurement of maternal plasma cysteinylated albumin may be useful for monitoring pregnancies associated with UPI and for detection of IUGR. PMID- 15791657 TI - Magnetic resonance image findings of placental lake: report of two cases. AB - Two cases with a large cystic mass within the placenta are reported. By ultrasonography, it was found that both women had a subchorionic hypoechoic lesion (11.0 x 4.8 x 4.0 cm and 6.6 x 3.7 x 2.2 cm, respectively) at 24 and 35 weeks of gestation, respectively. In both cases, turbulent blood flow generated by a pulsatile jet flow (pulse rate; 40 to 60 beats per minute) into the cystic lesion seen on real-time imaging and lesions being low intensity on T1-weighted and isointensity on T2-weighted magnetic resonance image suggested that they contained fresh maternal blood. In both cases, the sonolucency of the lesions did not change until cesarean deliveries of females, both of whom were small-for gestational-age infants (1940 g at 37 weeks and 2195 g at 37 weeks, respectively). Biochemical analysis of the fluid in the cystic lesion sampled during the cesarean section in the latter case confirmed that the fluid had originated from the maternal blood. These lesions histologically corresponded to large avillous areas surrounded by normal villi. Thus, a huge placental lake was diagnosed in both cases. PMID- 15791658 TI - Second-trimester maternal serum screening for Down syndrome in in vitro fertilization pregnancies. AB - OBJECTIVES: To examine whether second-trimester maternal serum triple marker screening results differ between in vitro fertilization (IVF) pregnancies and naturally conceived pregnancies. METHODS: Second-trimester maternal serum triple marker screening results from 88 IVF pregnancies were compared with 596 naturally conceived pregnancies (controls). Controls were matched to each IVF pregnancy by maternal age, gestational age and date of blood collection. All pregnancies in the study were known to have normal outcome. Multiple of the median (MoM) levels of human chorionic gonadotrophin (hCG), unconjugated estriol (uE3) and alpha fetoprotein (AFP), and the false-positive rate for Down syndrome were compared between the two groups. RESULTS: No statistically significant differences in analyte levels or in Down syndrome false-positive rate were observed between the IVF and naturally conceived pregnancies. CONCLUSION: IVF patients can be counselled about maternal serum triple marker screening in the same manner as patients with naturally conceived pregnancies. There is no evidence to support the general use of analyte correction factors in the interpretation of second trimester maternal serum screen results in IVF pregnancies. PMID- 15791659 TI - Prenatally detected trisomy 20 mosaicism. AB - BACKGROUND: Trisomy 20 is one of the more common mosaic trisomies detected on amniocentesis and presents with a normal outcome in over 90% of reported cases. Trisomic cells are almost never confirmed in newborn blood and are only rarely found in other fetal or placental samples. Nonetheless, some abnormal outcomes have been reported, including unexplained fetal demise, intrauterine growth restriction, and multiple congenital anomalies. Because of the lack of molecular studies on such cases, it is unknown whether the origin of trisomy or presence of uniparental disomy (UPD) could have some influence on outcome. METHODS: We present data on six cases of trisomy mosaicism, two detected by chorionic villous sampling (CVS) and four by amniocentesis (AF), submitted to our laboratory for molecular studies. RESULTS AND CONCLUSIONS: A meiotic origin of the trisomy could be confirmed in only one of these cases. In addition, uniparental disomy was excluded in all four cases for which parents were available for testing. The four cases with low levels of trisomy in amniotic fluid (0%, 10%, 11%, and 12%) were associated with a normal outcome. The remaining two cases of trisomy 20 had high levels of trisomy in amniotic fluid (96% and 58%) and had abnormal outcomes (developmental delay in one and stillbirth with IUGR and severe oligohydramnios in the other). Including previously published cases, there is a clear association with the level of trisomy and outcome, with only 4% abnormal outcomes when <40% trisomic cells were detected. Higher levels of trisomy were also observed in male fetuses as compared to female fetuses (p = 0.01); however, there were no sex differences in frequency of abnormal outcomes. PMID- 15791660 TI - The association of echogenic fetal lungs with trisomy 21. AB - OBJECTIVE: To evaluate the association between echogenic appearing lungs on ultrasonographic examination of the fetus and aneuploidy. METHODS: Cases in which echogenic lungs were prospectively identified on ultrasonographic examination were collected. Other abnormal ultrasonographic findings were documented, as were patient data pertinent to the risk of aneuploidy, such as maternal age and results of serum screening. The chromosomal complement of each identified case is reported. RESULTS: Five fetuses were identified with diffusely echogenic lungs on ultrasound examination over a two-year period. In three of the five cases, there were other ultrasound abnormalities, although in only one of these was a major congenital abnormality detected. In all three of these cases, the fetal chromosomal complement showed trisomy 21. In 1 of the 2 cases in which the echogenic lungs were an isolated finding, the fetus also was found to have trisomy 21. CONCLUSIONS: Bilateral and diffusely echogenic lungs appear to have an association with fetal trisomy 21. PMID- 15791661 TI - Triploidy identified through second-trimester serum screening. AB - OBJECTIVES: To describe the maternal serum marker patterns of triploid pregnancies and estimate the second-trimester prevalence of triploidy. METHODS: Forty-two cases of triploidy were identified in six serum screening programmes, five in the United Kingdom, one in Canada. This study describes the serum marker patterns, serum screening results for Down syndrome, trisomy 18 and open neural tube defects, and maternal age of these triploidy cases. The risk cut-off levels were > or = 1 in 250 for Down syndrome, > or =2.5 MoMs alpha-fetoprotein for open neural tube defects and > or =1:100 for trisomy 18 screening. The estimated second-trimester prevalence of triploidy was based on 22 triploidy cases ascertained in 599 934 pregnancies from three routine screening programmes, which attempted complete ascertainment of aneuploidy cases. RESULTS: The observed second-trimester rate of triploidy was 0.37 per 10 000 fetuses. Two different serum marker patterns were seen in triploid pregnancies, distinguished from each other by typically very high or very low levels of total hCG or free beta-hCG. The median maternal ages were respectively 33 years for triploidy with human chorionic gonadotrophin levels < 1.0 MoM, and 26 years for those with hCG levels > or =1.0 MoM. Fifty-seven percent of the pregnancies with a triploid fetus had a risk estimate > or =1:100 for trisomy 18 alone, 10% had an alpha-fetoprotein > or =2.5 MoM, 5% were screen positive for Down syndrome alone, and 19% had an increased risk or positive results for more than one anomaly. CONCLUSION: The simultaneous use of maternal serum tests designed to screen prenatally for Down syndrome, neural tube defects, and an increased risk of trisomy 18 resulted in a screen-positive result for 90% of pregnancies with triploidy. PMID- 15791662 TI - Prenatal diagnosis of a cystic type IV sacrococcygeal teratoma mimicking a cloacal anomaly: contribution of MR. AB - OBJECTIVE: To report a case of pelvic midline cystic mass in a female fetus without visibility of the rectum and which is not a cloacal anomaly. METHODS: Ultrasound (US) and magnetic resonance imaging (MRI) were performed respectively at 27 and 27.5 weeks' gestation and the findings of these examinations were compared with post-mortem examination after termination of pregnancy (TOP). RESULTS: US showed a pelvic midline cystic mass in a female fetus with mild enlargement of the left ureter and renal pelvis. MRI did not show the T1 hypersignal of meconium in the rectum. The analysis of MR findings were suggestive of the diagnosis of a sacrococcygeal teratoma. Pregnancy was terminated because of the possibility of severe sphincterial disorders and the fetopathological examination confirmed this diagnosis. CONCLUSION: This observation illustrates the diagnostic problems generated by the detection of a pelvic midline cystic mass in a female fetus. The absence of visibility of the T1 hypersignal of the meconium in the rectum is traditionally supposed to be highly suggestive of a cloaca, but may also be explained by the emptiness of the rectum, compressed by the mass. PMID- 15791663 TI - Lack of correlation between elevated maternal serum hCG during second-trimester biochemical screening and fetal congenital anomaly. AB - OBJECTIVE: Isolated elevations in midtrimester maternal serum human chorionic gonadotrophin concentrations (MShCG) have been reported to be associated with a substantially increased likelihood of fetal congenital malformations. The reported malformations included a wide range of organ systems, originating at different embryologic developmental stages. The purpose of our study was to determine the significance of an isolated elevated MShCG (>2.5 MoM) in midtrimester for the detection of fetal structural anomalies in a large population. METHODS: Among 10,144 women who underwent a biochemical triple screen at 15 to 18 weeks' gestation, 463 patients, who had an elevated MShCG, but normal alpha-fetoprotein (AFP) and unconjugated estriol (uE3) levels, were identified. Patients with an integrated calculated Down syndrome risk above 1:250 were excluded. Only nonsmokers, at ages <35 years, without a history of prior fetal anomalies were included. The control group consisted of 463 patients with normal serum analyte concentrations and Down syndrome risks below 1:250, who were matched for maternal age and date of biochemical screen. All patients underwent a detailed genetic sonogram in which an anatomic survey and multiple 'soft markers' for aneuploidy were looked for. Newborns were examined by a senior pediatrician trained in dysmorphology. RESULTS: MShCG levels were 3.18 +/- 0.72 versus 0.99 +/ 0.43 MoM (p < 0.0001) in study and control groups respectively. Sonography revealed 8 versus 6 cases of major congenital anomalies among the 463 patients of their respective groups, and 39 versus 36 sonographic 'soft markers' for aneuploidy. Fetal karyotyping and neonatal examination for dysmorphology revealed 6 chromosomal anomalies (4 Down syndrome; 2 Turner syndrome) among the 8 major malformations in the study group, but none in the controls (p < 0.0001). Three of the 39 fetuses with 'soft markers' and elevated MShCG were found to have trisomy 21. CONCLUSION: Isolated elevation of MShCG does not confer an increased risk of fetal congenital anomalies other than chromosomal abnormalities. However, elevated MShCG levels in combination with sonographic 'soft markers' for aneuploidy were associated with a high incidence of chromosomal anomalies, despite a normal biochemical triple screen risk estimate. PMID- 15791664 TI - Prenatal diagnosis of megacystis-microcolon-intestinal hypoperistalsis syndrome: contribution of amniotic fluid digestive enzyme assay and fetal urinalysis. AB - OBJECTIVES: Megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS) is a usually lethal disease during the first year of life. There is no specific ultrasound prenatal diagnosis and no identified genetic locus. The value of amniotic fluid digestive enzyme assay and fetal urine biochemistry in the prediction of MMIHS was analysed. METHODS: Retrospective study of 14 MMIHS cases. Amniotic fluid digestive enzymes and fetal urine biochemistry were compared in MMIHS and megabladder (63 and 264 cases respectively). RESULTS: Abnormal amniotic fluid digestive enzyme profile (vomiting of bile or digestive secretion leakage) was observed in 8/10 MMIHS cases. These profiles were observed in 7/63 controls; 80% sensitivity (95%CI = 55%-100%); 89% specificity (95%CI = 81%-96%). Fetal urinalysis was normal in 12/12 MMIHS cases except high calcium (>0.6 mmol/l). This profile was observed in 33/264 megabladder control cases; 100% sensitivity; 98.7% specificity (95%CI = 83.5%-91.5%). CONCLUSION: For the first time, we propose a prenatal diagnosis of MMIHS based on amniotic fluid digestive enzyme assay and on fetal urinalysis. PMID- 15791665 TI - Ectrodactyly-ectodermal dysplasia-clefting (EEC) syndrome presenting with a large nephrogenic cyst, severe oligohydramnios and hydrops fetalis: a case report and review of the literature. AB - OBJECTIVES: To report a case of EEC syndrome with a large nephrogenic cyst detected by prenatal ultrasonography. METHODS: Prenatal ultrasonographic detection, genetic counselling, termination of pregnancy, radiographic study, autopsy and ultrastructural study of scalp hair. The literature on EEC syndrome with genitourinary anomalies and prenatal diagnosis was also reviewed. RESULTS: A 6-cm cyst in the right side of abdominal cavity was detected in a fetus of 24 weeks' gestational age with severe oligohydramnios and hydrops fetalis. The autopsy revealed bilateral renal dysplasia with a large nephrogenic cyst at the right side and markedly hypoplastic urinary bladder and pulmonary hypoplasia. The fetus also had ectrodactyly and syndactyly of hands and feet and ectodermal dysplasia and left cleft lip and palate. Ultrastructure of hair from scanning electron microscopy revealed no obvious abnormality. CONCLUSION: This is a rare case of fetal malformation partly detected prenatally. In order to detect this syndrome, careful ultrasound search for acral anomalies should be performed in cases with renal malformation or obstructive uropathy. PMID- 15791666 TI - Developing a relativities approach to valuing the prevention of non-fatal work related accidents and ill health. AB - The aim of the current explorative study is to define and test a process for the valuation of the benefits associated with the prevention of non-fatal work related accidents and ill health. A relativities approach is adopted, and monetary values for the prevention of three forms of work-related illness are estimated. The approach involves describing relevant attributes of alternative events (accidents or occurrences of ill health), their causes, the characteristics of the relevant working population, and the number of events that are avoidable, and asking respondents to make pair wise choices between alternatives options for prevention. Indirect monetary valuations are obtained against a peg event for which a reliable valuation exists (road deaths).A series of discussion groups were held to identify relevant factors affecting potential valuations and to test the presentation of information. The predicted magnitude of responses for three-case study events (and road deaths) was estimated in a pilot study. These preliminary stages informed the final survey instrument that described five attributes in addition to a statement of the event and occupation, and the likely intervention effect, which was administered by post. Based on a small sample, the results show that virtually all respondents passed the inserted consistency test. The median respondent altered their choice according to the number of events avoided for all three comparisons, such that the estimated valuations appear sensible. Potential amendments are suggested, but the general relativities approach warrants further investigation for the valuation of non fatal work-related accidents and ill health. PMID- 15791667 TI - Comparison of two techniques for transcervical cell sampling performed in the same study population. AB - OBJECTIVES: The aim of this study was to evaluate and compare the presence of fetal cells in transcervical cell (TCC) samples collected in the first trimester of pregnancy by two different procedures [mucus collection and intrauterine lavage (IUL)], performed consecutively in the same subjects scheduled for elective termination of pregnancy (TOP). METHODS: A total of 126 mucus/IUL sample pairs were retrieved from pregnant women immediately before TOP at a gestational age ranging from 7 to 12 weeks; at termination, samples of placental tissue were collected in all cases. All mucus samples were analysed by a polymerase chain reaction (PCR) assay and, in a subset of experiments involving 56 specimens, also by fluorescence in situ hybridization (FISH) procedure. IULs were divided in two aliquots, one for PCR analysis and one for the preparation of FISH slides. All placental tissue samples obtained at termination were analysed by FISH for fetal sexing. The PCR assay for fetal sex determination was performed by using, in a multiplex reaction, primers for SRY (Y chromosome sex-determining region, 738 bp) and HUMARA (human androgen receptor on the X chromosome, 280 bp) genes. The FISH analysis was carried out using direct-labelled commercial probes for X chromosome alpha-satellite (DXZ1, Xp11.1-q11.1, spectrum green) and Y chromosome alpha satellite (DYZ3, Yp11.1-q11.1, spectrum orange) regions. RESULTS: In samples from known male pregnancies (n = 67), full concordance between IUL and mucus results could be found in 11 cases (16.4%); in 41 cases, Y chromosome material was detected by FISH (n = 2), by PCR (n = 5) or both (n = 34) in IUL samples, but not in the corresponding mucus samples. Y chromosome material was not documented in 10 mucus/IUL sample pairs. In 5 cases, the FISH (n = 2), the PCR (n = 1) or both (n = 2) failed to detect Y chromosome material in IULs, which was detected, however, by PCR in the corresponding mucus samples. Overall, correct sex prediction was achieved in 55/67 IULs (82%) and in 16/67 (23.9%) mucus samples from male pregnancies. In samples from known female pregnancies (n = 56), full concordance between results of IUL/mucus pairs and those on placental samples could be found in 53 cases (94.6%); in 3 cases, Y chromosome material was documented by PCR in mucus samples, but not in the corresponding IULs. Correct sex prediction was therefore achieved in 56/56 IULs (100%) and in 53/56 (94.6%) mucus samples from female pregnancies. CONCLUSION: This study provides evidence that, among TCC sampling techniques, IUL, but not mucus collection, can yield fetal cells in a constant and reliable fashion, which is a basic prerequisite for possible clinical usage. This suggestion had already emerged from some previous investigations but, owing to the study design, differences in study populations can no longer be used to explain the very different and sometimes-conflicting results reported in earlier studies. PMID- 15791668 TI - Prenatal diagnosis of trisomy 4p: a new locus for holoprosencephaly? AB - Trisomy of the short arm of chromosome 4 is a well-known syndrome, and several observations have been made in the last 30 years. Herein, we report a new observation of trisomy 4p in a fetus with a semi-lobar holoprosencephaly (HPE), dysmorphic features and multiple malformations. The diagnosis of HPE was made, at 33 weeks' gestation, on the fetus of a healthy G1P0 woman. Amniocentesis was performed for chromosome analysis and additional material was found on a chromosome 22. The couple elected to terminate the pregnancy and fetal examination was realized. Conventional and molecular cytogenetic studies were performed on the fetus and the parents, which showed that the additional material found on one chromosome 22 corresponded to the short arm of chromosome 4 and therefore led us to establish a diagnosis of trisomy 4p inherited from the malsegregation of a paternal translocation t(4;22)(q12;q11.1). The etiology of HPE is very heterogeneous; it includes non-genetic factors such as maternal diabetes and genetic causes. HPE cases have been described in association with many chromosomal anomalies, trisomy 13 being the most frequent. However, to our knowledge, HPE has never been previously reported in association with a trisomy involving solely the short arm of chromosome 4. PMID- 15791669 TI - Inequality decomposition and geographic targeting with applications to China and Vietnam. AB - How far are income-related inequalities in the health sector due to gaps between poor and less poor areas, rather than due to differences between poor and less poor people within areas? This note sets out a method for answering this question, and illustrates it with two empirical examples. The disproportionate accrual of health subsidies to Vietnam's better-off is found to be largely due to the fact that richer provinces have larger per capita subsidies, while pro-rich inequalities in health insurance coverage in rural China are found to be largely due to the fact that better-off villages have been more successful at preventing the collapse of their insurance schemes. PMID- 15791670 TI - Influence of laser wavelength and pulse duration on gas bubble formation in blood filled glass capillaries. AB - BACKGROUND AND OBJECTIVES: Hypervascular skin lesions (HVSL) are treated with medical lasers characterized by a variety of parameters such as wavelength lambda, pulse duration t(p), and radiant exposure E that can be adjusted for different pathology and blood vessel size. Treatment parameters have been optimized assuming constant optical properties of blood during laser photocoagulation. However, recent studies suggest that this assumption may not always be true. Our objective was to quantify thermally induced changes in blood that occur during irradiation using standard laser parameters. STUDY DESIGN/MATERIALS AND METHODS: Glass capillary tubes (diameter D = 100, 200, and 337 microm) filled with fresh or hemolyzed rabbit blood were irradiated once at lambda = 585, 595, or 600 nm, t(p) = 1.5 milliseconds; and also at lambda = 585 nm, t(p) = 0.45 milliseconds. E was increased until blood ablation caused formation of permanent gas bubbles. In a corroborative study, human blood was heated at 50 degrees C and absorbance spectra were measured as a function of time. RESULTS: Threshold radiant exposure, E(thresh), for gas bubble formation was found not to depend on lambda, which might be surprising in view of the 10 fold lower absorption coefficient at 600 nm as compared to 585 nm. The spectroscopic study revealed heat-induced changes in blood constituent composition of hemoglobins (Hb) from initially 100% oxyhemoglobin (HbO2) to deoxyhemoglobin (HHb) and, ultimately, methemoglobin (metHb) as the major constituent. Model calculations of E(thresh)(lambda,D) based on changing constituent blood composition during heating with milliseconds lasers were found to correlate with experimental results. CONCLUSIONS: For laser treatment of HVSL it appears that lambda is of secondary importance and that the choice of t(p) is a more important factor. PMID- 15791673 TI - Placental mesenchymal dysplasia associated with fetal aneuploidy. AB - OBJECTIVES: To describe three cases of placental mesenchymal dysplasia (PMD) associated with abnormal karyotype and review the cases reported in the literature. METHODS: The cases were retrieved from the files of three different institutions. A search of the English language literature was performed using Medline database. RESULTS: Placental abnormalities suggestive of molar changes were seen on the prenatal ultrasound scans. Histologically, the cases had large, hydropic stem villi with myxomatous stroma, cistern formation and 'chorangiomatoid' changes. The placental and fetal karyotypes identified were trisomy 13 (47,XX,t(1:13)(q32;q32)+ 13); Klinefelter syndrome (47,XXY) and triploidy (69,XXX). Including these 3 cases, of 66 reported cases, 51 (78%) were female and 14 (22%) male (ratio 3.6:1); the karyotype was normal in 32/36 (89%) and abnormal in 4/36 (11%); Beckwith-Wiedemann syndrome was confirmed or suspected in 15/66 (23%). Excluding termination of pregnancies, intrauterine death occurred in 18/54 (33%) cases. CONCLUSION: Molar ultrasonographic appearances associated with increased maternal serum alpha-fetoprotein but normal, or slightly elevated, levels of ss human Chorionic Gonadotrophin should raise the clinical suspicion of PMD. The diagnosis of this condition should not be disregarded when an abnormal fetus and/or an abnormal karyotype are demonstrated. PMID- 15791672 TI - Protein synthesis-dependent LTP in isolated dendrites of CA1 pyramidal cells. AB - Local dendritic protein synthesis provides a mechanism by which the effects of gene expression can be spatially restricted within synaptodendritic compartments of neurons. In the present study, we show that long-term potentiation (LTP), induced by two strong tetanic trains, can be produced in isolated CA1 apical dendrites and that new protein synthesis is critical for this dendritic LTP. LTP in isolated dendrites is also blocked by rapamycin, an inhibitor of growth related protein synthesis, similar to that observed for LTP in the intact hippocampal slice. These results show that increased dendritic translation of proteins is a critical molecular mechanism for hippocampal LTP. PMID- 15791671 TI - Helicobacter pylori in patients can be killed by visible light. AB - BACKGROUND: Helicobacter pylori colonizes the mucus layer of the human stomach and may cause peptic ulcer and adenocarcinoma. Novel antimicrobial approaches are sought due to the occurrence of antibiotic resistance and consequent treatment failure. We report here that H. pylori is susceptible to inactivation by blue light. STUDY DESIGN/MATERIALS AND METHODS: A controlled, prospective, blinded, trial of endoscopically delivered blue light to eradicate H. pylori in regions of the gastric antrum, in 10 patients between the ages of 21 and 80 who tested positive for H. pylori. Light (405 nm) (40 J/cm2) was delivered to a 1-cm diameter spot in the gastric antrum via optical fiber passed through the endoscope and weighed biopsies were taken from treated and control spots and colonies quantitatively cultured. RESULTS: Blue light killed 5 logs of bacteria in vitro. The mean reduction in H. pylori colonies per gram tissue between treated and control spots was 91% (7.4+/-4.8 x 10(6) vs. 8.1+/-1.9 x 10(7), two tailed P < 0.0001). Some patients had reductions approaching 99%. No differences were observed on histological examination of light-treated and control gastric tissue. CONCLUSION: Blue light phototherapy may represent a novel approach to eradication of H. pylori, particularly, in patients who have failed standard antibiotic treatment. PMID- 15791674 TI - GP reimbursement and visiting behaviour in Ireland. AB - In Ireland, approximately 30% of the population receive free GP services (medical card patients) while the remainder (private patients) must pay for each visit. In 1989, the manner in which GPs were reimbursed by the State for their medical card patients was changed from fee-for-service to capitation while private patients continued to pay on a fee-for-service basis. Concerns about supplier-induced demand were in part responsible for this policy change. The purpose of this paper is to examine the extent to which the utilisation of GP services is influenced by the reimbursement system facing GPs, by comparing visiting rates for the two groups before and after this change. Using a difference-in-differences approach on pooled micro-data from 1987, 1995 and 2000, we find that medical card eligibility has a consistently positive and significant effect on the utilisation of GP services. However, the differential in visiting rates between medical card patients and others did not narrow between 1987 and 1995 or 2000, as might have been anticipated if supplier-induced demand played a major role prior to the change in reimbursement system. PMID- 15791675 TI - The effect of Taiwan's tax-induced increases in cigarette prices on brand switching and the consumption of cigarettes. AB - The effect of raising cigarette taxes to reduce smoking has been the subject of several studies, which often treat the price of cigarettes as an exogenous factor given to smokers who respond to it by adjusting their smoking behavior. However, cigarette prices vary with brand and quality, and smokers can and do switch to lower-priced brands to reduce the impact of the tax on the cost of cigarettes as they try to consume the same number of cigarettes as they had before a tax hike. Using data from a two-year follow-up interview survey conducted before and after a new cigarette tax scheme was imposed in Taiwan in 2002, this study examines three behavioral changes smokers may make to respond to tax-induced cigarette price increase: brand-switching, amount consumed, and amount spent on smoking. These changes were studied in relation to smoker income, before-tax cigarette price, level of addiction, exposure to advertizing, and consumer loyalty. We found that smokers, depending upon exposure to advertizing, level of consumer loyalty and initial price of cigarettes, switched brands to maintain current smoking habits and control costs. We also found that the initial amount smoked and level of addiction, not price, at least not at the current levels in Taiwan, determined whether a smoker reduced the number of cigarettes he consumed. PMID- 15791676 TI - AIDS education, condom demand, and the sexual activity of American youth. AB - This paper examines the effects of AIDS education at school and at home on the sexual behavior of American youth. Multinomial logit equations of the probabilities of abstinence, sexual intercourse with a condom, and intercourse without a condom are estimated using data from the Youth Risk Behavior Supplement of the 1992 National Health Interview Survey. We find no significant effects of AIDS education on the probability of abstinence, but we do find that AIDS education significantly raises the likelihood of condom-protected relative to unprotected intercourse. These results indicate that risk-altering and risk revealing AIDS education dominate any utility-altering effects favoring intercourse over abstinence. We also find that young women are influenced by AIDS education to a greater extent than young men. Overall, our results suggest that educating young people about AIDS does not promote sex and encourages safer sex, reducing the likelihood of HIV transmission and lowering the subsequent social costs. PMID- 15791677 TI - Integration of imaging and pathological studies in Meckel-Gruber syndrome. PMID- 15791678 TI - Tobacco industry price-subsidizing promotions may overcome the downward pressure of higher prices on initiation of regular smoking. AB - Real cigarette prices in the US increased from the early 1980s to early 1990s. Holding all else equal, adolescent initiation of regular smoking should have declined during this period. Using national population-based surveys (n = 336 343) conducted in the 1990s, we present trends (early 1960s to mid-1990s) in the initiation of regular smoking among 14-17-year-old adolescents and 18-21-year-old young adults. We also present trends in consumer-price-index-adjusted cigarette price and tobacco-industry expenditures for price-subsidizing promotions. We relate price and price-subsidizing tobacco industry expenditures to trends in initiation in the two age groups, using autoregressive integrated moving average models (ARIMA). From the model results, we conclude that price-subsidizing promotions may provide the tobacco industry with an effective way to segment the market. That is, they effectively offer lower prices to population subgroups that are more price-sensitive (e.g. young smokers not yet addicted), countering the depressing effect of general price increases on smoking. Thus, we find that the relationship of cigarette price to smoking behavior is more complex than previously described. PMID- 15791679 TI - A propensity score approach to estimating the cost-effectiveness of medical therapies from observational data. AB - Health summary measures are commonly used by policy makers to help make decisions on the allocation of societal resources for competing medical treatments. The net monetary benefit is a health summary measure that overcomes the statistical limitations of a popular measure namely, the cost-effectiveness ratio. We introduce a linear model framework to estimate propensity score adjusted net monetary benefit. This method provides less biased estimates in the presence of significant differences in baseline measures and demographic characteristics between treatment groups in quasi-randomized or observational studies. Simulation studies were conducted to better understand the utility of propensity score adjusted estimates of net monetary benefits when important covariates are unobserved. The results indicated that the propensity score adjusted net monetary benefit provides a robust measure of cost-effectiveness in the presence of hidden bias. The methods are illustrated using data from SEER-Medicare for the treatment of bladder cancer. PMID- 15791680 TI - Recurrent fetal pyelectasis in a family with fetuses associated with partial trisomy 10q (10q24.1-->qter). PMID- 15791681 TI - Prenatal diagnosis of concomitant alobar holoprosencephaly and caudal regression syndrome associated with maternal diabetes. PMID- 15791682 TI - Long-term psychological consequences of pregnancy termination for fetal abnormality: a cross-sectional study. AB - OBJECTIVE: We examined women's long-term psychological well-being after termination of pregnancy (TOP) for fetal anomaly in order to identify risk factors for psychological morbidity. METHODS: A cross-sectional study was performed in 254 women, 2 to 7 years after TOP for fetal anomaly before 24 weeks of gestation. We used standardised questionnaires to investigate grief, posttraumatic symptoms, and psychological and somatic complaints. RESULTS: Women generally adapted well to grief. However, a substantial number of the participants (17.3%) showed pathological scores for posttraumatic stress. Low educated women and women who had experienced little support from their partners had the most unfavourable psychological outcome. Advanced gestational age at TOP was associated with higher levels of grief, and posttraumatic stress symptoms and long-term psychological morbidity was rare in TOP before 14 completed weeks of gestation. Higher levels of grief and doubt were found if the fetal anomaly was presumably compatible with life. CONCLUSION: Termination of pregnancy for fetal anomaly is associated with long-lasting consequences for a substantial number of women. Clinically relevant determinants are gestational age, perceived partner support, and educational level. PMID- 15791683 TI - Long term post-natal follow-up after transabdominal chorion villus sampling compared to amniocentesis. PMID- 15791684 TI - A panel data study of physicians' labor supply: the case of Norway. AB - Physicians are key personnel in a sector which is important due to its size as well as the quality of service it provides. We estimate the labor supply of physicians employed at hospitals in Norway, using personnel register data merged with other public records. A dynamic labor supply equation is estimated using a sample of 1303 male physicians observed over the period 1993-1997. The methods of estimation are GMM and system GMM. We reject the static model in favor of a dynamic model and obtain short run wage elasticities around 0.3. This is higher than previously estimated for physicians, in particular for those who are not self-employed. PMID- 15791685 TI - New studies on the Murchison meteorite shed light on the pre-RNA world. PMID- 15791686 TI - GPCR antitarget modeling: pharmacophore models for biogenic amine binding GPCRs to avoid GPCR-mediated side effects. AB - G protein-coupled receptors (GPCRs) form a large protein family that plays an important role in many physiological and pathophysiological processes. However, the central role that the biogenic amine binding GPCRs and their ligands play in cell signaling poses a risk in new drug candidates that reveal side affinities towards these receptor sites. These candidates have the potential to interfere with the physiological signaling processes and to cause undesired effects in preclinical or clinical studies. Here, we present 3D cross-chemotype pharmacophore models for three biogenic amine antitargets: the alpha(1A) adrenergic, the 5-HT(2A) serotonin, and the D2 dopamine receptors. These pharmacophores describe the key chemical features present within these biogenic amine antagonists and rationalize the biogenic amine side affinities found for numerous new drug candidates. First applications of the alpha(1A) adrenergic receptor model reveal that these in silico tools can be used to guide the chemical optimization towards development candidates with fewer alpha(1A) mediated side effects (for example, orthostatic hypotension) and, thus, with an improved clinical safety profile. PMID- 15791687 TI - A homologue of the Mycobacterium tuberculosis PapA5 protein, rif-orf20, is an acetyltransferase involved in the biosynthesis of antitubercular drug rifamycin B by Amycolatopsis mediterranei S699. PMID- 15791688 TI - Protein-detecting microarrays: current accomplishments and requirements. AB - The sequencing of the human genome has been successfully completed and offers the chance of obtaining a large amount of valuable information for understanding complex cellular events simply and rapidly in a single experiment. Interestingly, in addressing these proteomic studies, the importance of protein-detecting microarray technology is increasing. In the coming few years, microarray technology will become a significantly promising and indispensable research/diagnostic tool from just a speculative technology. It is clear that the protein-detecting microarray is supported by three independent but strongly related technologies (surface chemistry, detection methods, and capture agents). Firstly, a variety of surface-modification methodologies are now widely available and offer site-specific immobilization of capture agents onto surfaces in such a way as to keep the native conformation and activity. Secondly, sensitive and parallel detection apparatuses are being developed to provide highly engineered microarray platforms for simultaneous data acquisition. Lastly, in the development of capture agents, antibodies are now probably the most prominent capture agents for analyzing protein abundances. Alternative scaffolds, such as phage-displayed antibody and protein fragments, which provide the advantage of increasing diversity of proteinic capture agents, however, are under development. An approach involving recombinant proteins fused with affinity tag(s) and coupled with a highly engineered surface chemistry will provide simple production protocols and specific orientations of capture agents on the microarray formats. Peptides and other small molecules can be employed in screening highly potent ligands as well as in measuring enzymatic activities. Protein-detecting microarrays supported by the three key technologies should contribute in accelerating diagnostic/biological research and drug discovery. PMID- 15791689 TI - New insights for pursuing high relaxivity MRI agents from modelling the binding interaction of Gd(III) chelates to HSA. PMID- 15791690 TI - Development of an electrospray ionization mass spectrometric method for the quantification of theophylline in horse serum. AB - A rapid and selective method has been developed for the determination of theophylline in horse serum by LC-ESI/MS/MS. The analytical method includes a protein precipitation extraction for sample preparation, liquid chromatography separation technique and ionspray tandem mass spectrometry. The drug was extracted from serum using a protein precipitation with acetonitrile and the supernatants were analyzed using an LC-ESI/MS/MS instrument. The chromatography was performed using a 50 x 2.1 mm C(8) analytical column and an isocratic mobile phase composes of 60:40 acetonitrile-0.5% formic acid in water with a flow rate fixed at 350 microL/min. A linear (weighted 1/concentration) relationship was used to perform the calibration over an analytical range of 0.1-20 ppm. The intra batch precision and accuracy at LLOQ, medium and high concentration were 11.7, 6.9 and 5.4% and 95.8, 107.8 and 95.8%, respectively, and the inter-batch precision and accuracy at LLOQ, medium and high concentration were 10.4, 7.9 and 7.3% and 97.3, 105.2 and 95.9%, respectively. This LC-ESI/MS/MS method for the determination of theophylline in horse serum has been proved to within generally accepted criteria used for bioanalytical assay and was used successfully during clinical investigation. PMID- 15791691 TI - Interaction of the nuclear localizing cytolytic granule serine protease granzyme B with importin alpha or beta: modulation by the serpin inhibitor PI-9. AB - Conditional on perforin-dependent delivery to the nucleus of target cells, the cytolytic granule serine protease granzyme B (GrB) plays a central role in eliciting the nuclear events of apoptosis, as shown by the fact that reducing GrB nuclear entry prevents nuclear apoptosis. Apart from a requirement for cytosolic factors and lack of dependence on the guanine-nucleotide-binding protein Ran, little is known regarding the nuclear import pathway of GrB. In this study we use quantitative yeast two-hybrid and direct binding assays to show that GrB can be recognized independently by either of the nuclear import receptor family members importin (IMP) alpha and beta1, but that these proteins either alone or in combination cannot replace exogenous cytosol to reconstitute GrB nuclear import in vitro. Whereas antibodies to IMP(alpha) inhibit transport, indicating that IMP(alpha) is required for GrB nuclear import, those to IMP(beta) enhance transport, implying that IMP(beta) inhibits GrB nuclear import; consistent with this, the addition of recombinant IMP(beta) but not IMP(alpha) reduces maximal nuclear accumulation in the presence of cytosol. Intriguingly, complexation of GrB with its specific serpin inhibitor PI-9 was found to prevent recognition by IMP(beta) but not by IMP(alpha), and eliminate the apparent requirement for IMP(alpha) for nuclear import. We conclude that GrB nuclear import exhibits complex regulation by IMPs; that heterodimerization with PI-9 can modulate the interaction has implications for protection against apoptosis. PMID- 15791692 TI - Hematosalpinx: an unusual complication after medical abortion with oral mifepristone and misoprostol. PMID- 15791693 TI - Severe ductal constriction in the third-trimester fetus following maternal self medication with nimesulide. AB - OBJECTIVE: This was a multicenter series of 10 cases of constriction of the fetal ductus arteriosus occurring after maternal ingestion of one or two oral doses of nimesulide, which led to emergency delivery in four cases. METHODS: The computerized databases of three referral centers were searched for all cases of ductal constriction detected during the last 8 years in association with maternal ingestion of nimesulide. Ten cases were retrieved and represent the study population. The following variables were analyzed: gestational age at diagnosis, reason for nimesulide ingestion, total nimesulide dose, time interval between drug intake and ultrasound examination, indication for echocardiography, presence of right ventricular hypertrophy, presence of tricuspid valve regurgitation, ductal systolic and diastolic peak velocities, ductal pulsatility index, signs of fetal heart failure, pregnancy and feto-neonatal outcome, and ductal status at neonatal echocardiography. RESULTS: Nimesulide ingestion was the result of self medication in all cases. The mean gestational age at diagnosis was 36.3 weeks and the mean time interval between nimesulide intake and echocardiographic examination was 2.9 days. The five cases with ductal peak velocities > 2.5 m/s showed right ventricular dysfunction with significant tricuspid regurgitation. At echocardiography performed immediately after birth, the ductus appeared closed in all cases and two neonates showed persistent pulmonary hypertension requiring intermittent positive pressure ventilation and nitric oxide therapy for 1 week. CONCLUSIONS: Although the true incidence of the adverse effect of nimesulide ingestion during pregnancy cannot be deduced, we believe that patients and their clinicians should be alerted to the possibility that clinically significant ductal constriction may follow maternal self-medication with just one or two oral doses of nimesulide, at least if the drug is taken near term. PMID- 15791694 TI - Fetal cerebral ventriculomegaly: outcome in 176 cases. AB - OBJECTIVE: To evaluate the outcome of fetuses affected by different degrees of ventriculomegaly. METHODS: We studied 176 fetuses with ventriculomegaly and evaluated the pregnancy outcome and the neurodevelopmental outcome at age > or = 24 months. The population was divided into three groups according to ventricular width: A (mild ventriculomegaly, 10 to 12 mm); B (moderate, 12.1 to 14.9 mm) and C (severe, > or = 15 mm). RESULTS: Ventriculomegaly was more often an isolated finding in Group A (44/75; 58.7%) than in Group B (10/41; 24.4%) and Group C (24/60; 40%). When the ventriculomegaly was an isolated finding, 97.7% of fetuses with mild, 80% with moderate and 33.3% of those with severe dilatation were alive at > or = 24 months. The neurodevelopmental outcome was normal in 93% of Group A, 75% of Group B and 62.5% of Group C. CONCLUSIONS: Our results suggest that the definition of borderline ventriculomegaly should be limited to ventricular width below 12 mm. Cases with measurements above this value are more often associated with malformations and have a normal neurodevelopmental outcome less frequently. PMID- 15791695 TI - Psychological outcome in women undergoing termination of pregnancy for ultrasound detected fetal anomaly in the first and second trimesters: a pilot study. AB - OBJECTIVE: To ascertain and compare psychological morbidity following first- and second-trimester termination for fetal anomaly. METHODS: This was a cohort study of 30 women aged 20-40 years in a north London teaching hospital, 14 of whom had had a first-trimester termination and 16 a second-trimester termination for fetal anomaly. The main outcome measures were questionnaire data (General Health Questionnaire-28, Beck Depression Inventory, Perinatal Grief Scale, Impact of Event Scale (IES)) at 6 weeks, 6 months and 12 months after termination. RESULTS: There were high levels of psychological distress in both groups at each time point, and for the combined group the mean total scores on the IES remained above the cut-off for the entire study period. Those having second-trimester terminations had a significantly higher level of post-traumatic stress symptomatology 6 weeks after termination (14/16 vs. 6/14; odds ratio = 9.3; 95% CI, 1.5-57.7). CONCLUSIONS: Psychological morbidity following termination of pregnancy for fetal anomaly is prevalent and persistent. Our data suggest that in the short term (as assessed at a 6-week follow-up), second-trimester termination may be more stressful compared with first-trimester termination. PMID- 15791696 TI - Barriers to interferon-alpha therapy are higher in intravenous drug users than in other patients with acute hepatitis C. AB - BACKGROUND/AIMS: Treatment with interferon-alpha (IFN-alpha) may eradicate HCV in most acute hepatitis C patients, thus preventing chronic hepatitis and avoiding less efficacious combination therapy. METHODS: In a prospective study, we evaluated the impact of barriers to successful start and completion of treatment of acute and subacute (<12 months from infection) hepatitis C with pegylated IFN alpha2b, 1.5 microg/kg, QW, for 24 weeks. RESULTS: Out of 27 patients (22 were active intravenous drug users [IVDU]), 5 cleared HCV spontaneously. Antiviral treatment was indicated in 22 patients: six refused therapy for fear of side effects, whereas two others were lost to observation. Eight patients completed the treatment or received >80% of the scheduled drug: seven (88%) were sustained virological responders 24 weeks after the end of treatment. Six patients (all IVDU) stopped prematurely due to side effects: only one had a sustained virological response. Based on an intent-to-treat analysis, and considering all 14 patients in whom at least one dose of drug was administered, only 8 (57%) became sustained virological responders. CONCLUSIONS: Treatment of acute hepatitis C with pegylated IFN-alpha is highly beneficial, but its effectiveness is affected by a poor rate of acceptance and/or adherence to currently available regimens, especially in IVDU and women. PMID- 15791697 TI - Etanercept as an adjuvant to interferon and ribavirin in treatment-naive patients with chronic hepatitis C virus infection: a phase 2 randomized, double-blind, placebo-controlled study. AB - BACKGROUND/AIMS: Current therapies for patients with chronic hepatitis C virus (HCV) do not achieve sustained viral clearance in most patients, and are associated with severe toxic effects. Our aim was to investigate the efficacy and safety of etanercept as adjuvant to interferon and ribavirin in treatment-naive patients with HCV. METHODS: Double-blind, randomized, placebo controlled trial. Fifty patients with chronic HCV were randomly assigned to receive interferon alfa 2b and ribavirin with either etanercept or placebo for 24 weeks. The main outcome measure was the absence of HCV RNA at 24 weeks, the on treatment response at the end of the etanercept randomization period. RESULTS: At 24 weeks, HCV RNA was absent in 63% (12/19) etanercept patients compared to 32% (8/25) placebo patients (P=0.04). In addition, patients receiving etanercept had lower frequency of most adverse events categories compared to placebo. CONCLUSIONS: Etanercept given for 24 weeks as adjuvant therapy to interferon and ribavirin significantly improved virologic response at the end of the etanercept randomization period among patients with HCV, and was associated with decreased incidence of most adverse effects associated with interferon and ribavirin. PMID- 15791699 TI - [Recommendations for behavior in collaboration between the pharmaceutical industry with physicians]. PMID- 15791698 TI - [Psychosocial aspects of fertility disorders]. AB - The number of couples experiencing unwanted childlessness will in all likelihood continue to grow. Thus, ever more couples are undergoing IVF treatment; in Germany, this is, however, successful in only 13.9% of attempts. An unfulfilled wish for a child can have both negative emotional effects on individual partner and consequences for the couple's relationship. Women in particular suffer from the psychological stress that can be caused by infertility; they are more anxious, depressed, and have a decreased self-esteem than their partners. The desire to counteract these emotional strains and to enhance the quality of life is increasing and accordingly requests for counseling services are on the rise. As is the case in so many other psychosocial counseling services offered, there are shortcomings in the information available and a threshold of fear and dread of stigmatization by others persist. Studies have shown that various psychological treatments can often contribute to reducing stress but they do rarely increase the possibility of pregnancy. PMID- 15791700 TI - [Legal particulars in treatment of minor patients]. AB - According to current jurisdiction any intervention of the physical integrity of the body, even medical treatment and the administration of medication, constitutes a physical injury. The legal authority for this primarily comes from the consent of the patient. The problem for the medical doctor is whose consent is necessary when a patient is a minor. According to the jurisdiction and the literature, the consent of a minor to medical treatment is a legally binding decision not dependent on reaching the age of majority. Nevertheless, according to the overwhelming opinion it can be assumed that minors under the age of 14 years old are not yet capable of consent. The authority for consent lies with the parents or guardians. Because children are normally only accompanied by one of the parents when visiting a doctor, in routine practice the 3-stage theory developed by the Federal High Court pays a deciding role. According to this theory, for routine cases the doctor can assume that one of the parents is basically a representative of the other; for more complicated or severe cases the doctor must satisfy himself that this is true. For patients over 14 years old, the doctor must ascertain whether the patient is capable of consent. Even though a 15-year-old patient for example, can possess the necessary power of judgement and therefore be considered capable of consenting to routine measures or simple interventions, such as taking a blood sample, the yardstick for higher-risk operations, even routine ones is much higher. PMID- 15791701 TI - [Therapeutic approaches to acute pulmonary embolism]. PMID- 15791702 TI - Palifermin and chemotherapy-induced oral mucositis. PMID- 15791703 TI - Palifermin and chemotherapy-induced oral mucositis. PMID- 15791704 TI - Care of the potential organ donor. PMID- 15791705 TI - Cancer of the ovary. PMID- 15791706 TI - Perspectives. States scramble for ways to cover working uninsured. PMID- 15791707 TI - [To Chantal and Lucette]. PMID- 15791708 TI - There is a great shortage of midwives. PMID- 15791709 TI - The 'on-call' debate. PMID- 15791710 TI - Kidney, liver transplant study for people with HIV. AB - A study of liver and kidney transplants for persons with HIV, at 19 U.S. transplant centers, is open to new patients. Those who may need a transplant later might benefit by getting into the system in advance to avoid delays. PMID- 15791711 TI - HIV "new strain" story: for more information. AB - AIDS community groups have brought together background documents, explanatory writeups, and other information about the media stories that resulted from what is still a single, ambiguous case. PMID- 15791712 TI - Early Medicaid treatment: bipartisan bill in Senate with 32 cosponsors, could cut HIV deaths on Medicaid in half. AB - A bill to allow states to treat HIV early under Medicaid, instead of waiting for disabling illness, could prevent half of the HIV deaths in that program. PMID- 15791713 TI - Tat inhibitors, a new approach: interview with Olaf Kutsch, Ph.D. Interview by John S. James. AB - An important potential target for antiretrovirals is the HIV protein Tat (produced by the virus and essential for infection, but is not used by the human body). Years ago, a Tat inhibitor worked well in the laboratory but failed in patients. Modern biotechnology may have shown why--and how to screen for drugs more likely to work. PMID- 15791714 TI - If the condom breaks: new U.S. guidelines for non-occupational exposure to HIV. AB - Finally there are U.S.-government guidelines for prevention of non-occupational HIV exposure, for example after rape or accident. PMID- 15791715 TI - Global good news: many more treated. AB - A combination of financing from rich countries, determination and effort by local governments, effective teamwork, and successful scale-up of treatment access despite obstacles, has doubled the number of people receiving treatment in sub Saharan Africa (and also in East, South, and Southeast Asia) in six months. PMID- 15791717 TI - Epidemic continues to stabilize, except for black females, MSMs. New testing, prevention efforts being tried. AB - The CDC reports that recent data show a stable number of HIV diagnoses in 2003, although diagnoses among men who have sex with men have risen. PMID- 15791716 TI - FDA advisory on nevirapine. AB - The FDA summarized existing warnings against ongoing nevirapine treatment in women with a CD4 count over 250, due to a greatly increased risk of serious liver toxicity. (The warnings do not apply to single-dose nevirapine, which does not cause this problem.) PMID- 15791718 TI - CDC's prevention initiative shows no testing increase. Various demonstration projects funded. AB - Officials with the CDC expect to see some results from the HIV Prevention Initiative in 2004, although the data remain to be analyzed and the data from the 2003 HIV Prevention Initiative in 2003 showed it was having no impact, but officials are hopeful 2004 data will be different. PMID- 15791719 TI - Repeated testing advised for at-risk MSM groups. Some may need testing more than once a year. PMID- 15791720 TI - Social network enlists community recruiters. Targeted testing has produced 8% positive rate. AB - The Center for Multicultural Wellness and Prevention in Orlando, FL, has identified 14 HIV-positive cases out of 173 people tested in its first year of a demonstration project targeting black and Hispanic populations. PMID- 15791722 TI - Have tents, will travel in San Francisco. Testing facility travels to clients. AB - The San Francisco Tenderloin area has an estimated 1,200 people who are HIV positive and not aware of it. So a group is offering rapid HIV testing in a climate-controlled tent to encourage participation. PMID- 15791721 TI - Harlem center focuses on prevention for positives. Risk-reduction strategies are main focus. AB - Harlem, NY, organization recognizes people who engage in high-risk activities won't change behaviors overnight, so it focuses on reducing risk behaviors one step at a time. PMID- 15791723 TI - Rapid HIV testing popular with Chicago CBO clients. Testing has found 2% positive rate. AB - A Chicago organization provides rapid HIV testing in non-clinical settings to tap into hard-to-reach high-risk populations, including homeless youths and others. PMID- 15791724 TI - Rural HIV service center needs a creative approach. HIV testing has been done in karaoke bar. AB - A Southwest Louisiana group has found some creative solutions to the problems HIV staff have with trying to identify high-risk rural populations for HIV testing and counseling. PMID- 15791725 TI - World health community focuses on problems of women, HIV and violence for 2005. Women comprise nearly half of epidemic. PMID- 15791726 TI - Region shows how an effective program works. Mbeya region has declining trend of HIV. PMID- 15791727 TI - Medicare and Medicaid programs; fire safety requirements for certain health care facilities; amendment. Interim final rule with comment period. AB - This interim final rule with comment period adopts the substance of the April 15, 2004 temporary interim amendment (TIA) 00-1 (101), Alcohol Based Hand Rub Solutions, an amendment to the 2000 edition of the Life Safety Code, published by the National Fire Protection Association (NFPA). This amendment will allow certain health care facilities to place alcohol-based hand rub dispensers in egress corridors under specified conditions. This interim final rule with comment period also requires that nursing facilities install smoke detectors in resident rooms and public areas if they do not have a sprinkler system installed throughout the facility or a hard-wired smoke detection system in those areas. PMID- 15791729 TI - Memorial Hermann creates standalone patient communications system. AB - This standalone patient communications system can be customized by individual clinics. It also offers patients a series of message types, each of which can be routed to a different person at the clinics. And, though it required a security infrastructure, it didn't have to be integrated with backend systems. PMID- 15791728 TI - Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. AB - Prokaryotes contain short DN repeats known as CRISPR, recognizable by the regular spacing existing between the recurring units. They represent the most widely distributed family of repeats among prokaryotic genomes suggesting a biological function. The origin of the intervening sequences, at present unknown, could provide clues about their biological activities. Here we show that CRISPR spacers derive from preexisting sequences, either chromosomal or within transmissible genetic elements such as bacteriophages and conjugative plasmids. Remarkably, these extrachromosomal elements fail to infect the specific spacer-carrier strain, implying a relationship between CRISPR and immunity against targeted DNA. Bacteriophages and conjugative plasmids are involved in prokaryotic population control, evolution, and pathogenicity. All these biological traits could be influenced by the presence of specific spacers. CRISPR loci can be visualized as mosaics of a repeated unit, separated by sequences at some time present elsewhere in the cell. PMID- 15791730 TI - Real-time studies on microalgae under microgravity. AB - Using remote sensing technique, we investigated real-time Nostoc sphaeroides Kutz (Cyanobacterium) in Closed System under microgravity by SHENZHOU-2 spacecraft in January 2001. The experiments had 1 g centrifuges in space for control and ground control group experiments were also carried out in the same equipments and under the same controlled condition. The data about the population growth of Nostoc sp. of experiments and temperature changes of system were got from spacecraft every minute. From the data, we can find that population growth of Nostoc sp. in microgravity group was higher than that of other groups in space or on ground, even though both the control 1 g group in space and 1 g group on ground indicated same increasing characteristics in experiments. The growth rate of 1.4 g group (centrifuged group on ground) was also promoted during experiment. The temperature changes of systems are also affected by gravity and light. Some aspects about those differences were discussed. From the discussion of these results during experiment, it can be found that gravity is the major factor to lead to these changes. PMID- 15791731 TI - The roles of humans and robots in exploring the solar system. AB - Historically, advocates of solar system exploration have disagreed over whether program goals could be entirely satisfied by robotic missions. Scientists tend to argue that robotic exploration is most cost-effective. However, the human space program has a great deal of support in the general public, thereby enabling the scientific element of exploration to be larger than it might be as a stand-alone activity. A comprehensive strategy of exploration needs a strong robotic component complementing and supporting human missions. Robots are needed for precursor missions, for crew support on planetary surfaces, and for probing dangerous environments. Robotic field assistants can provide mobility, access to scientific sites, data acquisition, visualization of the environment, precision operations, sample acquisition and analysis, and expertise to human explorers. As long as space exploration depends on public funds, space exploration must include an appropriate mix of human and robotic activity. PMID- 15791732 TI - Overview of the Martian radiation environment experiment. AB - Space radiation presents a hazard to astronauts, particularly those journeying outside the protective influence of the geomagnetosphere. Crews on future missions to Mars will be exposed to the harsh radiation environment of deep space during the transit between Earth and Mars. Once on Mars, they will encounter radiation that is only slightly reduced, compared to free space, by the thin Martian atmosphere. NASA is obliged to minimize, where possible, the radiation exposures received by astronauts. Thus, as a precursor to eventual human exploration, it is necessary to measure the Martian radiation environment in detail. The MARIE experiment, aboard the 2001 Mars Odyssey spacecraft, is returning the first data that bear directly on this problem. Here we provide an overview of the experiment, including introductory material on space radiation and radiation dosimetry, a description of the detector, model predictions of the radiation environment at Mars, and preliminary dose-rate data obtained at Mars. PMID- 15791733 TI - Heavy ion observations by MARIE in cruise phase and Mars orbit. AB - The charged particle spectrum for nuclei from protons to neon, (charge Z=10) was observed during the cruise phase and orbit around Mars by the MARIE charged particle spectrometer on the Odyssey spacecraft. The cruise data were taken between April 23, 2001 and mid-August 2001. The Mars orbit data were taken March 5, 2002 through May 2002 and are scheduled to continue until August 2004. Charge peaks are clearly separated for charges up to Z=10. Especially prominent are the carbon and oxygen peaks, with boron and nitrogen also clearly visible. Although heavy ions are much less abundant than protons in the cosmic ray environment, it is important to determine their abundances because their ionization energy losses (proportional to Z2) are far more dangerous to humans and to instruments. Thus the higher charged nuclei make a significant contribution to dose and dose equivalent received in space. Results of the charged particle spectrum measurements will be reported. PMID- 15791734 TI - Solar particle events observed at Mars: dosimetry measurements and model calculations. AB - During the period from March 13, 2002 to mid-September, 2002, six solar particle events (SPE) were observed by the MARIE instrument onboard the Odyssey Spacecraft in Martian Orbit. These events were observed also by the GOES 8 satellite in Earth orbit, and thus represent the first time that the same SPE have been observed at these separate locations. The characteristics of these SPE are examined, given that the active regions of the solar disc from which the event originated can usually be identified. The dose rates at Martian orbit are calculated, both for the galactic and solar components of the ionizing particle radiation environment. The dose rates due to galactic cosmic rays (GCR) agree well with the HZETRN model calculations. PMID- 15791735 TI - A space radiation shielding model of the Martian radiation environment experiment (MARIE). AB - The 2001 Mars Odyssey spacecraft was launched towards Mars on April 7, 2001. Onboard the spacecraft is the Martian radiation environment experiment (MARIE), which is designed to measure the background radiation environment due to galactic cosmic rays (GCR) and solar protons in the 20-500 MeV/n energy range. We present an approach for developing a space radiation-shielding model of the spacecraft that includes the MARIE instrument in the current mapping phase orientation. A discussion is presented describing the development and methodology used to construct the shielding model. For a given GCR model environment, using the current MARIE shielding model and the high-energy particle transport codes, dose rate values are compared with MARIE measurements during the early mapping phase in Mars orbit. The results show good agreement between the model calculations and the MARIE measurements as presented for the March 2002 dataset. PMID- 15791736 TI - Findings of scientific misconduct. PMID- 15791737 TI - John Robert Vane: 1927-2004. PMID- 15791738 TI - Chemical meningitis after cerebral aneurysm treatment using two second-generation aneurysm coils: report of two cases. AB - OBJECTIVE AND IMPORTANCE: In the quest for effective and durable endovascular aneurysm treatment, second-generation aneurysm coils endeavor to increase the biological healing response to the implanted material. We report two cases of large cerebral aneurysms treated concurrently with both available second generation aneurysm coils and the subsequent development of symptomatic nonbacterial meningitis. CLINICAL PRESENTATION: Two previously healthy patients underwent endovascular treatment for large (>=2 cm) cerebral aneurysms. Both aneurysms were treated using multiple Hydrogel coils (MicroVention, Inc., Aliso Viejo, CA) and Matrix coils (Boston Scientific/Target, Fremont, CA). Careful aseptic technique was observed throughout each procedure, and prophylactic intravenous antibiotics were administered during the perioperative period to both patients. Treatment proceeded uneventfully in both cases with excellent aneurysm occlusion and no immediate postoperative neurological deficits. INTERVENTION: In both cases, the patients were discharged from hospital but quickly were readmitted with stigmata of meningitis. Imaging demonstrated durable occlusion of the aneurysms in both patients and also abnormalities indicative of perianeurysmal and diffuse intracranial inflammatory response. Complete septic workup failed to identify an organism in either patient. Both patients responded to treatment with corticosteroid medication used to modulate the inflammatory response induced by the coil implants. CONCLUSION: Second-generation aneurysm coils were developed to promote more durable occlusion of cerebral aneurysms by promoting more complete volumetric aneurysm occlusion or by eliciting a more prolific inflammatory response. The concurrent use of Hydrogel and Matrix coil systems in large aneurysms may cause an exuberant inflammatory response with both local and systemic manifestations. Although vigilant evaluation and treatment for presumptive bacterial meningitis is required in all such cases, patients respond to immunomodulatory therapy with corticosteroids. More information to understand better the interaction of Hydrogel and Matrix coils is needed. PMID- 15791739 TI - Progressive growth of a giant dolichoectatic vertebrobasilar artery aneurysm after complete Hunterian occlusion of the posterior circulation: case report. AB - OBJECTIVE AND IMPORTANCE: Dolichoectatic vertebrobasilar artery aneurysms are often extremely difficult, if not impossible, to treat with microneurosurgical clip reconstruction. As such, a Hunterian strategy via vertebral or basilar artery sacrifice is often used. We have encountered a patient in whom deliberate bilateral vertebral artery sacrifice was insufficient to avoid progressive expansion of a giant dolichoectatic vertebrobasilar artery aneurysm. On the basis of a review of the literature, we are unaware of another reported case. CLINICAL PRESENTATION: A 60-year-old man presented with signs and symptoms of brainstem compression from a large fusiform aneurysm involving the distal dominant vertebral and proximal basilar arteries. Results of angiographic evaluation were highly characteristic of underlying dolichoectasia. INTERVENTION: The patient was treated initially with staged bilateral vertebral artery occlusion and adjunctive posterior circulation revascularization. After this therapy failed, he underwent a trapping procedure and aneurysm deflation. CONCLUSION: Unclippable aneurysms of the vertebrobasilar system are formidable lesions. They are not uniformly treatable by direct surgical reconstruction, and their growth is not consistently stabilized by the implementation of a complete Hunterian strategy. Future developments related to the use of endovascular stent technology may offer a more successful treatment approach for patients with these complex cerebrovascular lesions. PMID- 15791740 TI - Traumatic venous varix causing sciatic neuropathy: case report. AB - OBJECTIVE AND IMPORTANCE: Sciatic neuropathy rarely presents in nonpenetrating trauma because of protection of the nerve by the pelvis, the gluteal muscles, and the tissues in the posterior thigh. We present the case of a patient who fell and subsequently developed a traumatic venous varix of the inferior gluteal vein that caused compression sciatic neuropathy. CLINICAL PRESENTATION: Seven days after a fall onto her right buttock, the patient developed a painful burning paresthesia in her leg and numbness on the dorsum of her foot. Numerous studies ruled out lumbar spine pathological abnormalities as the cause of the pain. Conventional magnetic resonance imaging revealed a lesion adjacent to the sciatic nerve. Gradient echo and two-dimensional time-of-flight magnetic resonance imaging sequences confirmed this to be a vascular lesion originating from the inferior gluteal vein and compressing the sciatic nerve. INTERVENTION: Operative resection obliterated the venous varix, thereby relieving the patient's pain and neurological deficit. CONCLUSION: No case of a traumatic venous varix of the inferior gluteal vein compressing the sciatic nerve has been reported to date. Surgical resection was successful in obliterating the lesion and relieving the symptoms. PMID- 15791741 TI - Temporary balloon occlusion of the cavernous carotid artery for removal of an orbital and intracranial foreign body: case report. AB - OBJECTIVE AND IMPORTANCE: We describe the first reported use of temporary balloon occlusion of the cavernous internal carotid artery for controlled removal of a foreign object from the cavernous sinus. This endovascular approach may be an alternative to craniotomy in highly selected cases. CLINICAL PRESENTATION: A 34 year-old incarcerated male attempted suicide by stabbing the earpiece of his glasses through his right orbit into the intracranial compartment. He presented with complete ophthalmoplegia. The earpiece traversed the cavernous sinus, penetrating its posterior wall to enter the perimesencephalic cistern and cerebellum. Angiography demonstrated a small direct carotid-cavernous fistula. INTERVENTION: Removal of the foreign body was performed under general anesthesia in the angiography suite with the operating room on standby. Nondetachable and detachable balloons were inflated in the cavernous carotid artery to provide vascular control while the foreign body was withdrawn from the cranium at the orbit. Follow-up angiographic runs with the balloons deflated revealed minimal arteriovenous shunting, which disappeared on subsequent studies. The balloons were removed. The patient remained neurologically stable with his baseline right ophthalmoplegia and V1-V2 hemianesthesia. At the 6-week follow-up, the patient remained clinically stable with no evidence of carotid-cavernous fistula or interval abscess formation. CONCLUSION: Endovascular temporary balloon occlusion of the cavernous carotid artery provides immediate control of the vessel (with an option of permanent carotid sacrifice), allowing removal of a foreign body without craniotomy in appropriate cases. PMID- 15791742 TI - Three-dimensionally reconstructed magnetic source imaging and neuronavigation in pediatric epilepsy: technical note. AB - OBJECTIVE: To determine the role of reconstructing three-dimensional magnetic source imaging (MSI) data on cortical resections for children undergoing epilepsy surgery using neuronavigation. METHODS: Magnetoencephalographic recordings were analyzed in 16 children under 18 years of age with intractable epilepsy. The data were transferred to the neuronavigation workstation for intraoperative localization of MSI spike sources in selected patients. With the aid of neuronavigation, the MSI spike sources were resected. Intraoperative electrocorticography was then used to survey the surrounding field for residual epileptiform activity. RESULTS: MSI spike sources were obtained in 13 of 16 patients. MSI spike sources localized the cortical and subcortical discharges before intraoperative electrocorticography in nine patients and before extraoperative subdural grid electroencephalographic monitoring in four patients. The localization of MSI spikes sources was characterized by clustered spike sources in 10 patients. By use of neuronavigation, the clustered spike sources were correlated to the interictal zone indicated by intraoperative electrocorticography in six patients and to the ictal onset zone shown on extraoperative subdural grid electroencephalography in three patients. Cortical excision of the spike cluster focus was then performed in these six patients. The technique used here to resect MSI spike source clusters that correlate with the ictal onset zone by invasive subdural grid monitoring is illustrated in one patient who underwent cortical resection for epilepsy surgery. CONCLUSION: Three dimensional reconstruction of MSI data linked to neuronavigation is a promising technique to facilitate resections around eloquent cortex in children with epilepsy. PMID- 15791743 TI - Preoperative localization of the carotid bifurcation for carotid endarterectomy using magnetic resonance angiography without tomography: technical note. AB - OBJECTIVE: Carotid endarterectomy without angiography is becoming increasingly common. Magnetic resonance imaging accurately demonstrates vascular anatomy and the degree of stenosis but may not localize the cervical carotid bifurcation for incision planning without tomography. We report a method for preoperative localization of the bifurcation using the relatively constant axial coplanar relationship between the horizontal petrous (C2) segment of the internal carotid artery and the external auditory canal (EAC). METHODS: The vertical distance between the midpoint of the C2 segment and the cervical carotid bifurcation was measured on magnetic resonance angiography images in five patients. Before surgery, a transverse incision was planned the same distance below the EAC. After exposure, the distance from the EAC to the bifurcation was measured with calipers. The distances from the anterior, posterior, superior, and inferior wound margins during retraction were also measured. RESULTS: The mean vertical distance between the C2 segment and the bifurcation was 7.2 cm on preoperative imaging and 7.6 cm between the EAC and bifurcation using intraoperative measurement (difference, 0.4 cm). The average distances between the midportion of the bifurcation and the wound margins after retractor placement were 2.9, 2.4, 2.8, and 3.5 cm for anterior, posterior, superior, and inferior measurements, respectively. CONCLUSION: The C2 segment-cervical carotid bifurcation distance may be used to estimate accurately the vertical distance from the EAC to the bifurcation. When coupled with a transverse incision for carotid endarterectomy, the use of this preoperative measurement produces a target well centered within the operative field. PMID- 15791744 TI - Seeding of a cavernous angioma with Mycoplasma hominis: case report. PMID- 15791745 TI - Operative techniques and instrumentation for neurosurgery. PMID- 15791747 TI - Idiopathic intracranial hypertension. PMID- 15791746 TI - Risk of postoperative hemorrhage after intracranial surgery after early nadroparin administration: results of a prospective study. PMID- 15791748 TI - Novel neuroscience M.D.-Ph.D. programs in Germany: a chance for neurosurgical research? PMID- 15791749 TI - An experimental comparison of telephone and personal health interview surveys. PMID- 15791750 TI - Chapter I. Study design. PMID- 15791751 TI - Chapter II. Differences between the telephone and personal interview data. PMID- 15791752 TI - Chapter III. Experimental interviewing techniques. PMID- 15791753 TI - Chapter IV. The effects of respondent rules on health survey reports. PMID- 15791754 TI - Chapter V. A comparison of CATI and non-CATI questionnaires. AB - It appears likely that a greater proportion of surveys will use the telephone as a medium of sampling and data collection in the future. As software developments proceed and computer hardware costs shrink, many of these will use CATI systems. This experiment can be used as a benchmark for the transition to CATI because it provides documentation on potential problems with changing modes. For most criteria, there are only small differences between CATI and non-CATI interviewing in this project. The criteria include response rates, reactions of the interviewer and respondent, and most health statistics of interest. There are, however, some exceptions to this finding of equivalence between methods. The first exception is the result that the average number of minutes per CATI interview exceeded that for non-CATI interviews. There also is some evidence that the interviewer variability estimates tend to be lower in CATI than non-CATI. Finally, there is evidence of lower skip error problems in the CATI interviews. The first of these results affects survey costs; the second and third, survey error. The first may be a function of software or hardware choice and thus can be addressed in new CATI designs. The second and third will be of benefit to all CATI systems in the future. PMID- 15791755 TI - Chapter VI. Measurement of interviewer errors in the SRC Telephone Survey. AB - The most important finding of this section is that unusually low levels of interviewer effects were measured in the telephone survey. This result may be due to the stringent controls on interviewer behavior that were introduced in this study but were absent in past studies. These low interviewer variances inhibited attempts to explain interviewer variability on the health variables. Because there was little interviewer variability, the correlates among the monitoring data were weak, and the findings did not exhibit consistency over variables. PMID- 15791756 TI - Chapter VII. Nonsampling bias and variance in the SRC Telephone Survey data. AB - This small exercise was a check on the nature of the effects of the experimental interviewing behaviors, one that attempted to dismiss the possibility that the increases in reporting were coming at the expense of greater interviewer variance. The findings presented are limited by the small number of statistics examined, but they suggest that the possibility that the experimental behaviors merely move errors from bias terms to variance terms is unlikely to be experienced. It is also unlikely that the experimental effects do not uniformly decrease interviewer variance and they appear to be reducing response bias. The measurement of the net effect of the treatments on mean square error requires estimates of the relative size of total response variance composed both of interviewer and respondent variability and response bias associated with the experimental procedures. PMID- 15791757 TI - Current estimates from the National Health Interview Survey, 1994. PMID- 15791758 TI - Summary health statistics for U.S. adults: National Health Interview Survey, 2001. AB - OBJECTIVES: This report presents health statistics from the 2001 National Health Interview Survey for the civilian noninstitutionalized adult population classified by sex, age, race and Hispanic origin, poverty status, and region of residence for chronic condition prevalence, health status and functional limitations, health care access and utilization, health behaviors, and human immunodeficiency virus (HIV) testing. Also, health statistics by education, income, health insurance coverage, marital status, and place of residence are presented for health status and limitations in activity, health care access and utilization, health behaviors, and knowledge and attitudes toward HIV. SOURCE OF DATA: The National Health Interview Survey is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics and is representative of the civilian noninstitutionalized U.S. population. Sociodemographic data are collected for everyone in the family during face-to-face interviews with adults present at the time of the interview. The health information for adults in this report was obtained from one randomly selected adult per family. HIGHLIGHTS: In 2001, 64% of adults 18 years of age and over reported excellent or very good health. Fifty-eight percent of adults never participated in any type of vigorous leisure-time physical activity, and 13% of adults did not have a usual place of health care. Twelve percent of adults had been told by a doctor or health professional that they had heart disease, and 21% had been told on two or more visits that they had hypertension. Nearly a quarter of all adults were current smokers, and 22% were former smokers. Based on estimates of body mass index, 36% of adults were overweight and 23% were obese. PMID- 15791759 TI - Inpatient utilization of short-stay hospitals by diagnosis United States, 1978. AB - Statistics are presented on the utilization of non-Federal short-stay hospitals based on data abstracted in the National Hospital Discharge Survey from a national sample of hospital records of discharged inpatients. The number of discharges, discharge rates, and average length of stay are shown for the classes and categories of first-listed diagnoses, by demographic characteristics of inpatients discharged and by geographic region and size of the hospitals. For these patient and hospital characteristics, this report also presents the number and percent distribution of all-listed diagnoses (up to five diagnoses per patient) reported for inpatients discharged. PMID- 15791760 TI - Early childhood chronic illness: comparability of maternal reports and medical records. AB - OBJECTIVES: The research reported here seeks to quantitatively assess the comparability of two widely used sources of information on child health: maternal reports and medical records. The analysis provides a comparison of how well maternal reports and medical provider data agree on 15 types of chronic health conditions, ranging from specific illnesses such as asthma or sickle cell anemia to broader categories such as chronic heart or orthopedic conditions, to impairments such as vision, hearing, or speech problems. METHODS: This study uses data on a nationally representative sample of 6201 preschool aged children whose mothers participated in the 1991 Longitudinal Followup (LF) to the 1988 National Maternal and Infant Health Survey (NMIHS) and whose identified health care providers supplied medical visit data for the children. The LF and NMIHS were conducted by the National Center for Health Statistics (NCHS), Centers for Disease Control and Prevention (CDC), in collaboration with a number of other agencies of the Federal Government. In the 1991 LF survey, the mothers' questionnaire included questions on chronic and acute health conditions, accidents requiring medical attention, and hospital admissions for their children that had been included in the 1988 NMIHS. The mothers were also asked to grant NCHS permission to request medical records from the children's medical providers. RESULTS: Kappa statistics reveal poor agreement for 12 of 15 conditions studied. Weighted prevalence estimates vary widely across sources. For some conditions, despite apparently similar prevalence estimates from the two sources, mothers' reports and medical records identified very different groups of children as "cases." Concurrence rates were lower for children from poor, less educated, and Hispanic families. Reasons for and implications of these findings are also discussed. PMID- 15791761 TI - Compensating for provider nonresponse using response propensities to form adjustment cells: the National Immunization Survey. AB - OBJECTIVES: The National Immunization Survey (NIS) uses two phases of data collection to obtain vaccination information from a sample of young children: a random-digit-dialing (RDD) survey for identifying households with children 19-35 months of age, followed by a mail survey for obtaining provider-reported vaccination histories about these children. Provider-reported vaccination histories are used to estimate vaccination coverage rates. In 1998, provider reported vaccination histories were not obtained for 32.9% of children with a completed RDD interview. This report describes the statistical methods adopted in 1998 to reduce the bias in vaccination coverage estimates that could result from "vaccination history nonresponse," that is, differences between children for whom provider data was obtained and those for whom it was not obtained. METHODS: In the methods adopted in 1998, children with completed NIS RDD interviews are grouped into adjustment cells defined by their propensity to have adequate provider data. Sampling weights of children with adequate provider data are divided by the cell-specific weighted response rate to allow these children to represent all children in the cell. RESULTS: Using an "optimal" number of cells, the overall extent of bias reduction was 0.5%, suggesting that provider nonresponse bias was small. Authoritative literature suggests using five cells. No statistically significant differences were observed in vaccination coverage estimates when comparing estimates based on the "optimal" number of cells with five cells. Thus, five adjustment cells are used to reduce provider nonresponse bias in the NIS vaccination coverage estimates. No substantively important differences were observed between estimates based on the methodology used prior to 1998 and the methodology adopted in 1998. PMID- 15791762 TI - Summary health statistics for U.S. children: National Health Interview Survey, 2001. AB - OBJECTIVES: This report presents statistics from the 2001 National Health Interview Survey (NHIS) on selected health measures for children under 18 years of age, classified by sex, age, race, Hispanic origin, family structure, parent's education, family income, poverty status, health insurance coverage, residence, region, and health status. The topics covered are asthma, allergies, learning disability, Attention Deficit Hyperactivity Disorder (ADHD), prescription medication, respondent-assessed health status, school-loss days, usual place of health care, time since last contact with a health care professional, unmet dental need, time since last dental contact, and selected measures of health care access. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention, National Center for Health Statistics, and is representative of the civilian noninstitutionalized population of the United States. Data are collected during face-to-face interviews with adults present at the interview. Information about children is collected for one randomly selected child per family in face-to-face interviews with an adult proxy respondent familiar with the child's health. SELECTED HIGHLIGHTS: In 2001, most U.S. children under 18 years of age enjoyed excellent or very good health (84%). However, 10% had no health insurance coverage, and 5% had no usual place of health care. Thirteen percent of children had ever been diagnosed with asthma. Eight percent of children 3-17 years of age had a learning disability, and 6% of children had ADHD. Lastly, 11% of children in single-mother families had two or more visits to an emergency room in the past year compared with 6% of children in two-parent families. PMID- 15791763 TI - Summary health statistics for U.S. adults: National Health Interview Survey, 2002. AB - OBJECTIVES: This report presents health statistics from the 2002 National Health Interview Survey (NHIS) for the civilian noninstitutionalized adult population, classified by sex, age, race and Hispanic origin, education, income, poverty status, health insurance coverage, marital status, place of residence, and region of residence for chronic condition prevalence, health status, functional limitations, health care access and utilization, health behaviors, and human immunodeficiency virus testing. The presentation of percentages and percent distributions in both age-adjusted and unadjusted versions is new this year. SOURCE OF DATA: The NHIS is a household, multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics. In 2002, data were collected for 31,044 adults for the Sample Adult questionnaire. The conditional response rate was 84.4%, and the final response rate was 74.4%. The health information for adults in this report was obtained from one randomly selected adult per family. HIGHLIGHTS: In 2002, 62% of adults 18 years of age or over reported excellent or very good health. Fifty-nine percent of adults never participated in any type of vigorous leisure-time physical activity, and 14% of adults did not have a usual place of health care. Eleven percent of adults had been told by a doctor or health professional that they had heart disease, and 21% had been told on two or more visits that they had hypertension. Twenty-two percent of all adults were current smokers, and 23% were former smokers. Based on their body mass index, 35% of adults were overweight, and 23% were obese. PMID- 15791764 TI - Introduction: Communicating critical test results. AB - The need for improving the timeliness of the communication of critical test results is reflected in a Joint Commission National Patient Safety Goal and is the basis for this special issue of the Journal. PMID- 15791765 TI - Doing better with critical test results. AB - Health care should achieve the goals that no critical test result is lost and that all such results are managed with a speed appropriate to their urgency. PMID- 15791766 TI - Communicating critical test results: safe practice recommendations. AB - BACKGROUND: Massachusetts hospitals have collaborated in a patient safety initiative conducted by the Massachusetts Coalition for the Prevention of Medical Errors and the Massachusetts Hospital Association which is aimed at improving the ability to communicate critical test results in a timely and reliable way to the clinician who can take action. Solutions to this problem would address enhancing communication, teamwork, and information transfer, all fundamental system factors linked to patient safety. DEVELOPING THE SAFE PRACTICE RECOMMENDATIONS AND THE "STARTER SET": A Coalition-convened Consensus Group defined critical test results as values/interpretations for which reporting delays can result in serious adverse outcomes for patients. The scope included laboratory, cardiology, radiology, and other diagnostic tests in inpatient, emergency, and ambulatory settings. The Consensus Group developed Safe Practice Recommendations to promote successful communication of results, and a "starter set" of test results sufficiently abnormal to be widely agreed to be considered "critical." DISSEMINATION: The recommendations and the starter set of test results were disseminated in a statewide collaborative open to all Massachusetts hospitals. Hospitals' team members tested changes and shared successful strategies that improved the reliability of communicating critical test results. An evaluation of the results of this collaborative is underway. PMID- 15791767 TI - Issues and initiatives in the testing process in primary care physician offices. AB - BACKGROUND: Errors occur frequently in management of the testing process in primary care physicians' offices. These errors may result in significant harm to patients and lead to inefficient practice. Important issues are summarized for primary care clinicians and their offices toconsider in improving the management of the testing processes. METHODS: To identify published efforts to improve management of the testing process, a literature search was performed and the references from the identified articles were checked for additional studies. Descriptive studies, expert opinion pieces, and controlled trials were all included. Unpublished results of ongoing studies in laboratory testing errors in primary care practice are presented. RESULTS: A conceptual model of the testing process was developed, with identified general and specific errors that occur in the testing process. On the basis largely of descriptive studies, ways are described to reduce testing process errors and the harm resulting from these errors. CONCLUSIONS: Standardization of processes, computerized test tracking systems (especially those embedded in electronic medical records), and attention to human factors issues are likely to reduce errors and harm. These ideas need confirmation in well-designed randomized trials and quality improvement initiatives. PMID- 15791768 TI - Failure to recognize and act on abnormal test results: the case of screening bone densitometry. AB - BACKGROUND: Failure to follow up on abnormal test results is common. A model was developed to capture the reasons why providers did not take action on abnormal test results. METHODS: A systematic review of the medical literature was conducted to identify why providers did not follow up on test results. The reasons were then synthesized to develop an operational model. The model was tested by reviewing electronic medical records of consecutive patients diagnosed with osteoporosis through a dual-energy x-ray absorptiometry (DXA) scan to determine whether: (1) the scan results had been reeviewed; (2) therapy was recommended; (3) the scan results were not reviewed and why this occurred. RESULTS: Of the 48 newly diagnosed osteoporosis patients, 16 did not receive a recommendation to begin treatment. There was no evidence that the scan results wrere reviewed in 11 of the 16 cases (23% of all abnormal scans); the scan results of an additional 5 patients were reviewed but no treatment was recommended. DISCUSSION AND CONCLUSIONS: A clinically significant ercentage of DXA scan results went unrecognized. As a long-term solution, direct patient notification could theoretically reduce the burden on providers, activate and empower patients, and create a back-up system for ensuring that patients are notified of their test results. PMID- 15791769 TI - Using a healthcare matrix to assess patient care in terms of aims for improvement and core competencies. AB - BACKGROUND: In 2001, the Institute of Medicine (IOM) recommended six Aims f or Improvement; the dimensions of quality describe a health care system that is safe, timely, effective, efficient, equitable, and patient centered. In 1999, the Accreditation Council of Graduate Medical Education (ACGME) adopted six core competencies that physicians in training must master if they are to provide quality care. A Healthcare Matrix was developed that links the IOM aims for improvement and the six ACGME Core Competencies. The matrix provides a blueprint to help residents to learn the core competencies in patient care, and to help faculty to link mastery of the competencies with improvement in quality of care. HEALTHCARE MATRIX: The Healthcare Matrix is a conceptual framework that projects an episode of care as an interaction between quality outcomes and the skills, knowledge, and attitudes (core competencies) necessary to affect those outcomes. For example, an anesthesiology resident used the Healthcare Matrix for a complex 18-hour episode of care with a life-threatening situation. ONGOING WORK AND RESEARCH AGENDA: Collecting and analyzing a series of matrices provides the foundation for systematic change in patient care and medical education and a rich source of data for operational and improvement research. PMID- 15791770 TI - Diagnostic errors in medicine: a case of neglect. AB - BACKGROUND: Medical diagnoses that are wrong, missed, or delayed make up a large fraction of all medical errors and cause substantial suffering and injury. Compared with other types of medical error, however, diagnostic errors receive little attention-a major factor in perpetuating unacceptable rates of diagnostic error. Diagnostic errors are fundamentally obscure, health care organizations have not viewed them as a system problem, and physicians responsible for making medical decisions seldom perceive their own error rates as problematic. The safety of modem health care can be improved if these three issues are understood and addressed. SOLUTIONS: Opportunities to improve the visibility of diagnostic errors are evident. Diagnostic error needs to be included in the normal spectrum of quality assurance surveillance and review. The system properties that contribute to diagnostic errors need to be systematically identified and addressed, including issues related to reliable diagnostic testing processes. Even for cases entirely dependent on the skill of the clinician for accurate diagnosis, health care organizations could minimize errors by using system-level interventions to aid the clinician, such as second readings of key diagnostic tests and providing resources for clinical decision support. Physicians need to improve their calibration by getting feedback on the diagnoses they make. Finally, clinicians need to learn about overconfidence and other innate cognitive tendencies that detract from optimal reasoning and learning. CONCLUSION: Clinicians and their health care organizations need to take active steps to discover, analyze, and prevent diagnostic errors. PMID- 15791771 TI - Medication errors: experience of the United States Pharmacopeia (USP). AB - This article in this new department introduces USP's error-reporting programs and efforts to help health care practitioners reduce the occurrence and severity of medication errors. PMID- 15791772 TI - Summary health statistics for the U.S. population: National Health Interview Survey, 2002. AB - OBJECTIVES: This report presents both age-adjusted and unadjusted health statistics from the 2002 National Health Interview Survey (NHIS) for the civilian noninstitutionalized population of the United States, classified by age, sex, race and Hispanic or Latino origin, family income, poverty status, education, place of residence, region of residence, and, where appropriate, health insurance coverage. The topics covered are health status and limitations in activities, special education or early intervention services, injuries and poisonings, health care access and utilization, and health insurance coverage. SOURCE OF DATA: The NHIS is a household, multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics. In 2002, household interviews were completed for 93,386 persons living in 36,161 households, reflecting a household response rate of 89.6%. SELECTED HIGHLIGHTS: Nearly 7 in 10 persons were in excellent or very good health in 2002. About 34 million persons (12%) were limited in their usual activities due to one or more chronic health conditions, and about 4 million persons (2%) required the help of another person with activities of daily living. About 6% of children received special education or early intervention services. Among persons under age 65 years, about 40 million (17%) did not have any health insurance coverage. The most common reason for lacking health insurance was cost, followed by a change in employment. PMID- 15791773 TI - [Vascular access guidelines for hemodialysis]. AB - Quality of vascular access (VA) has a remarkable influence in hemodialysis patients outcomes. Dysfunction of VA represents a capital cause of morbi mortality of these patients as well an increase in economical. Spanish Society of Neprhology, aware of the problem, has decided to carry out a revision of the issue with the aim of providing help in comprehension and treatment related with VA problems, and achieving an homogenization of practices in three mayor aspects: to increase arteriovenous fistula utilization as first vascular access, to increment vascular access monitoring practice and rationalise central catheters use. We present a consensus document elaborated by a multidisciplinar group composed by nephrologists, vascular surgeons, interventional radiologysts, infectious diseases specialists and nephrological nurses. Along six chapters that cover patient education, creation of VA, care, monitoring, complications and central catheters, we present the state of the art and propose guidelines for the best practice, according different evidence based degrees, with the intention to provide help at the professionals in order to make aproppiate decissions. Several quality standars are also included. PMID- 15791774 TI - [Pharmacogenomics]. PMID- 15791775 TI - [Drug therapy and pharmacogenomics--hopes and facts]. AB - Recent achievements in molecular genetics, in particular the characterization of a large number of non-synonymous single-nucleotide polymorphisms (ns-SNPs) have led to a substantial accumulation of knowledge about the mechanisms of interindividual variability in drug response. The concept of elucidating drug reactions by genetic analysis is termed "pharmacogenomics". Pharmacogenomics is set to provide a relevant contribution to our understanding of adverse drug reactions and non-responsiveness to drug therapy and may help realize the vision of providing individualized therapies. The greatest challenges for the field of pharmacogenomics relate to the issues of genotype-phenotype- and genotype environment- interactions, optimal selection of study designs, predictivity and ethical aspects. Although its current impact on the routine of medicine is minimal, it is likely that pharmacogenomics will provide important contributions to the practice of drug therapy. Pharmacogenetic concepts may prove important for approximately 20% of therapeutic drugs. PMID- 15791776 TI - [Therapeutic implications of ACE-gene polymorphism]. AB - The serum concentration of the angiotensin converting enzyme (ACE) depends on the individual genotype. Thus, ACE levels might be higher (DD-genotype), intermediate (ID) or low (II). Pre-therapeutic determination of the genotype might therefore optimize therapy with ACE-inhibitors or angiotensin II blockers. However, although numerous interventional studies have been performed they are hampered by a low number of included patients and have often yielded differing results. Thus, in order to overcome current controversies larger multicenter studies are warranted before recommending a general testing for genotype prior to initiation of therapy which interferes with the rennin-angiotensin-aldosterone system. PMID- 15791777 TI - [Therapeutic implications of polymorphisms in cytochromes and drug transporters]. AB - There is great heterogeneity in the way individuals respond to drug therapy. Reasons for this variability include pathophysiological or environmental factors, drug interactions or genetic influences. Among those influences are polymorphisms in drug-metabolizing enzymes, such as Cytochrom-P-450 (CYP) 2D6, CYP2C9 or CYP2C19 or genetic variants in enzymes coding for drug transporters, such as P Glycoprotein. Polymorphisms might cause changes in drug pharmacokinetics and consequently drug efficacy after administration of recommended standard drug doses. Reduced enzyme activity can either result in a higher percentage of drug side-effects, or an augmented drug response due to increased target site concentrations. Conversely, intensified catalytic enzymatic activity might lead to subtherapeutic drug concentrations. In the future, genetic screening by genotyping before the initiation of pharmacotherapy might help to identify responders or non-responders and might it offer individualized therapies to select patient populations. PMID- 15791778 TI - ["Applied kinesiology" in medicine and dentistry--a critical review]. AB - The "Applied Kinesiology" evolved in the USA is increasingly being used in Central Europe. In this review the development of the method and its practical application in medicine and in dentistry are elucidated. Furthermore the propagation of the method by the International College of Applied Kinesiology (= ICAK) and the "Touch for Health" (=TFH) is described. School medicine's criticism of "Applied Kinesiology" as well as the methodological replies from "Applied Kinesiologists" are outlined. It is important to realise that there is to date a lack of evidence for the effectiveness, validity and reliability of "Applied Kinesiology". The following requirements are thus vital: "Applied Kinesiologists" must develop clear criteria for single subgroups of "Applied Kinesiology", prove the effectiveness of their methods, and explain their findings in agreement with current medical knowledge. PMID- 15791779 TI - [A body mass index related scale for reconstructive breast reduction]. AB - PURPOSE: Breast reduction is a highly emotional theme and bears conflicting interest groups: 1) women who are suffering from symptomatic macromastia and therefore would wish to have their breast reduction paid by the insurers, irrespective of the amount of resection weight, 2) the insurance companies, who are ready to cover only really medically indicated operations and due to a lack of objective parameters often apply the very strict, arbitrary criterium for a minimum resection weight of 500 g per breast and 3) the surgeons who try to provide a fair, scientific basis for the differentiation between cosmetic and reconstructive indications for breast reductions for the sake of both the patients and the insurance parties. Concerned about such a generalizing rule we undertook a retrospective review of our patients' charts with both, cosmetic and reconstructive indications to judge the available, more-level minimum resection weight standards and see wether they were appropriate to use, or to provide an objective and measurable guideline for a scaled amount of breast reduction beyond the 500 g-resection-rule, adapted to the individual woman's body proportions. METHODS: 136 women could be included in the study. The resection weight was recorded and correlated to various parameters of the body proportions such as weight, height, the body mass index (BMI) and the body surface. The results were compared to the available minimum resection weight rules. RESULTS: The resection weight ranged from 55 to 1530 g (mean 450 g +/- 266 g, median 406 g). Overweight was present in 36% of all patients, whereas obesity was present in 7.5% of women. The mean BMI was 25.1 kg/m2. Of the twenty-four patients (18%), who were classified a priori as having a cosmetic indication, 4 (18%) had more than 500 g breast tissue resected bilaterally. On the other hand, in 55% of reconstructive patients less than the predicted 500 g of breast tissue had been resected. From all examined parameters the BMI had the highest correlation to the resected mean breast tissue (r = 0.64, p = < 0.001). DISCUSSION: Our retrospective review thus showed that with an arbitrary 500 g breast resection-rule all women beyond the mean values for weight and height were clearly put at a disadvantage. Also not completely solving this problem are the already available, more objective guidelines for graded minimum resection weight recommendations, which have relied on the body weight or the body surface area, parameters that both had a much lower correlation to the resected breast tissue in the patient group than the BMI. We therefore suggest using the BMI as the basis for a graded, more-level weight resection standard for reconstructive breast reductions. This algorithm is related solely to objectifying data and thus avoids biases from empirically derived data or hardly quantifiable breast (or obesity)-related pain syndromes, and respects all the different body builds of women. PMID- 15791780 TI - [Modern ceramic-on-ceramic bearings in total hip arthroplasty]. AB - OBJECTIVE: Ceramic bearings in total hip arthroplasty were already employed three decades ago, but due to high failure rates, their application decreased. However, improved material and design have led to their regaining popularity. The present study evaluates the short-term results of a modern ceramic-on-ceramic bearing in total hip prosthesis. METHODS: 229 total hip arthroplasties, alpha Cera Fit, were implanted in 227 patients over a 12-month period. A follow-up, at a minimum of 36 months, was performed. In addition to demographics, the Harris Hip-Score was used to calculate the clinical results and radiological control was also carried out. RESULTS: The mean age at the time of surgery was 64.6 (+/- 11.7) years. There was no case of deep infection, four patients had superficial wound healing disorders. 197/227 patients with 199 total hip arthroplasties were available for follow-up, with a mean follow-up time of 42 (+/- 2.9) months. The mean Harris Hip-Score was 94.3 (+/- 6.8). Radiological evaluation did not reveal any signs of lysis or loosening. Massive periarticular calcification was seen in three patients, with one that needed surgical debridement. CONCLUSION: The presented modern ceramic-on ceramic bearings in total hip arthroplasty show excellent clinical and radiological short-term results. The material-related complications reported in former publications were no longer seen. PMID- 15791781 TI - Whose life is it anyway--the dying patient's or the nurse's? PMID- 15791782 TI - Documentation of individualized patient care: a qualitative metasynthesis. AB - The aim of this study was to increase understanding of how individual patient care and the ethical principles prescribed for nursing care are implemented in nursing documentation. The method used was a metasynthesis of the results of 14 qualitative research reports. The results indicate that individualized patient care is not visible in nurses' documentation of care. It seems that nurses describe their tasks more frequently than patients' experiences of their care. The results also show that the structure of nursing documentation and the forms or manner of recording presupposed by the organization may prevent individual recording of patient care. In order to obtain visibility for good patient-centred and ethical nursing care, an effort should be made to influence how the content of nursing care is documented and made an essential part of individual patient care. If the content of this documentation does not give an accurate picture of care, patients' right to receive good nursing care may not be realized. PMID- 15791783 TI - Meeting ethical challenges in acute nursing care as narrated by registered nurses. AB - Five registered nurses were interviewed as part of a comprehensive investigation by five researchers into the narratives of five enrolled nurses (study 1, published in Nursing Ethics 2004), five registered nurses (study 2) and 10 patients (study 3) describing their experiences in an acute care ward at one university hospital in Sweden. The project was developed at the Centre for Nursing Science at Orebro University Hospital. The ward in question was opened in 1997 and provides care for a period of up to three days, during which time a decision has to be made regarding further care elsewhere or a return home. The registered nurses were interviewed concerning their experience of being in ethically difficult care situations in their work. Interpretation of the theme 'ethical problems' was left to the interviewees to reflect upon. A phenomenological hermeneutic method (inspired by the French philosopher Paul Ricoeur) was used in all three studies. The most prominent feature revealed was the enormous responsibility present. When discussing their responsibility, their working environment and their own reactions such as stress and conscience, the registered nurses focused on the patients and the possible negative consequences for them, and showed what was at stake for the patients themselves. The nurses demonstrated both directly and indirectly what they consider to be good nursing practices. They therefore demand very high standards of themselves in their interactions with their patients. They create demands on themselves that they believe to be identical to those expected by patients. PMID- 15791784 TI - Patients with cancer: their approaches to participation in treatment plan decisions. AB - The aim of this study was to explore experiences of participation in treatment planning decisions from the perspective of patients recently treated for colorectal cancer. Ten patients were purposively selected and interviewed. Constant comparative analysis, the core concept of grounded theory, was used. The dimensions were developed and organized into the main theme of 'compliant participation in serious decisions', which was composed of the two variations: complying with participation; and complying without participation. Complying with participation was characterized by feelings of self-confidence and self competence and by open dialogue between the participants, significant others and the physician. Complying without participation was characterized by participants' feelings of uncertainty and distress, and of being rushed into submitting to decisions without having time to reflect on the information provided or the opportunity to influence the treatment and care process. To participate (or choosing not to participate) builds on open and affirming dialogue, information and knowledge about the illness. Patient participation in treatment and care decision making is interpreted as a health promoting way of coping with illness. PMID- 15791785 TI - Does privacy matter? Former patients discuss their perceptions of privacy in shared hospital rooms. AB - As a relative concept, privacy is difficult to define in universal terms. In the New Zealand setting recent legislation aims to protect patients' privacy but anecdotal evidence suggests that these policies are not well understood by some providers and recipients of health care. This qualitative study set out to identify some of the issues by exploring former patients' perceptions of privacy in shared hospital rooms. The findings suggest a conditional acceptance of a loss of privacy in an environment dictated by architectural structure and by fiscal and time constraints. Participants indicated an awareness that personal information could be overheard and that their preference for a choice of setting for serious discussions was desirable. Some enjoyed the support offered in shared rooms, while, for others, overhearing another person's health issues caused unnecessary distress. The participants suggested that knowing they could be overheard constrained information disclosure. This withholding of information has implications for health professionals' ability to diagnose and treat patients appropriately. PMID- 15791786 TI - Teaching health care ethics: why we should teach nursing and medical students together. AB - This article argues that teaching medical and nursing students health care ethics in an interdisciplinary setting is beneficial for them. Doing so produces an education that is theoretically more consistent with the goals of health care ethics, can help to reduce moral stress and burnout, and can improve patient care. Based on a literature review, theoretical arguments and individual observation, this article will show that the benefits of interdisciplinary education, specifically in ethics, outweigh the difficulties many schools may have in developing such courses. PMID- 15791787 TI - Ethics, government and sexual health: insights from Foucault. AB - The work of Michel Foucault, the French philosopher who was interested in power relationships, has resonated with many nurses who seek a radically analytical view of nursing practice. The purpose of this article is to explore 'ethics' through a Foucauldian lens, in a conceptual and methodological sense. The intention is to provide a useful framework that will help researchers critically to explore aspects of nursing practice that relate to the construction of the self, morality and identity, be that nurse or patient related. The fundamentals of the research method of genealogy and the methods of ethics are reviewed. Using an example taken from the sexual health practice area, advice is given on how to structure data collection, incorporate interview data, avoid discourse determinism and measure resistance. PMID- 15791788 TI - What should HIV/AIDS be called in Malawi? AB - HIV/AIDS is the leading cause of morbidity and mortality in the southern African country of Malawi. At the largest referral health facility in Blantyre, the Queen Elizabeth Central Hospital, the majority of patients hospitalized in medical wards and up to a third of those in the maternity unit are infected with HIV. Many patients in the surgical wards also have HIV/AIDS. Health professionals in Blantyre, however, often choose not to write down the diagnosis of HIV or AIDS; rather, they prefer to use 'SGOT', 'ELISA' and 'spot test' to represent the HIV test, while 'immunosuppression', '?CD4 disease' and 'ARC' are preferred instead of 'AIDS'. It is possible that health professionals' belief that mentioning HIV and/or AIDS will harm patients is encouraging them to use these euphemisms. The use of less than exact terms to label HIV and AIDS may not be without cost. For instance, future attempts to conduct retrospective case study research may be hampered by this practice, which is not in accordance with the international classification of diseases. It is suggested that, although stigmatization and discrimination could be important driving factors in the use of cryptic language, it may be more worthy to fight discrimination and stigmatization head-on, rather than create avenues where these reactions may be perpetuated. PMID- 15791789 TI - War, nursing and morality. PMID- 15791790 TI - A call for education. PMID- 15791791 TI - Medical ethics at Ege University Faculty of Medicine. PMID- 15791792 TI - Evaluation of bromate ions level introduced by sodium hypochlorite during post disinfection of drinking water. AB - During the last ten years, interest concerning the occurrence of bromate in drinking water has grown due to its potential carcinogenicity and the new regulations. One source of bromate in finished water is due to its presence in the sodium hypochlorite solutions used for the disinfection of water. In fact, the brine solutions used for the production of sodium hypochlorite contain bromide ions in varying degrees that subsequently generate a certain quantity of bromate ions. Bromate concentrations ranging from 82 to 857 mg l(-1) (0.5-7.4 mg BrO3-/ g Cl2) have been found in commercial solutions of sodium hypochlorite used by Societe Anonyme Gestion des Eaux de Paris (SAGEP), a company that produces drinking water for Paris, France. In addition, the chlorine concentration of the hypochlorite solution can decrease during storage, consequently the added amount of bromate increases for a given applied dose of chlorine. PMID- 15791793 TI - Influence of cadmium on the performance of an activated SBR sludge treatment. AB - High concentration of heavy metals is a toxic factor for most microorganisms. As a result, such metals give rise to severe disruption in wastewater treatment operations. It has been noticed, however, that biological systems may adapt to heavy metals when their concentration is limited. In fact, such systems can even produce some degree of metal elimination provided that an optimal exposure tempo is guaranteed. On the other hand, irreversible damage may occur when metal concentration is sufficiently high. It is the aim of the present study to report on the inhibiting effects due to Cadmium on the activated sludge treatment method. To that end, different Cd concentrations were tested (ranging from 0.25 to 14 mg l(-1)) in a Sequential Biological Reactor, using synthetic wastewater without recirculation. The feed water providing 325 mg l(-1) COD and 30 mg l(-1) NH4+ -N. The process was screened by checking elimination percentages for COD and ammonia, as well as the evolution of ciliate protozoa. In addition, research was carried out on Cd assimilation by the activated sludge system by means of measuring in the mixed liquor and in the effluent. Results showed a marked decrease in purification efficiency when Cd-concentration rose over 14 mg l(-1), in which case COD elimination dropped from 98% to 41% while ammonia elimination went down from 98% to 32%. The Sludge Biotic Index decreases to 0 at Cd concentrations above 10 mg l(-1). PMID- 15791794 TI - Effect of some operational parameters on the decolorization of textile effluents and dye solutions by ozonation. AB - Treatment of three wastewaters and two acid dye solutions by ozonation with varying ozone dose, initial pH, and initial dye concentration were examined. Ozonation of all the wastewaters were shown to be pseudo-first-order reaction with respect to color, chemical oxygen demand (COD) and dissolved organic carbon (DOC). Increasing ozone dose increased the removal of color, COD, and DOC, as well as decreased ozone consumption ratio, and increased apparent rate constants. Ozonation was more successful at decolorization than the removal of COD and DOC or augmentation of biodegradability. Increasing initial dye concentration increased the ozone consumption and decreased the rate constant. Color removal efficiencies for different ozone dose application were in the range of 60-91% for sample A (dye bath effluent), and 70-96% for sample B (plant effluent). For sample A, COD and DOC removal efficiencies at several ozone dose applications were between 13-22 % and 8-13%, respectively. For sample B, they were between 11 33% and 6-15% respectively. Increasing pH could expand the efficiency of ozone slightly. Ozone consumption per unit color, COD and DOC removal at any time was found to bealmost the same whereas the applied ozone dose was different at a constant pH. Ozonation increased the biochemical oxygen demand (BOD,) of sample A and B 1.22 and 1.37 times by consuming 300 and 225 mg ozone, respectively. Ozonation of acid dyes was a pseudo-first order reaction with respect to dye. Increases in dye concentration raised specific ozone consumption. Specific ozone consumption for Acid Red 183 (AR-183) dye solution with 50 mg l(-1) of concentration rose from 0.32 to 0.72 mg-O3 per mg dye decomposed as the dye concentration was increased to 500 mg l(-1). PMID- 15791795 TI - A driving cycle for vehicle emissions estimation in the metropolitan area of Mexico City. AB - A driving cycle derived from driving behavior and real traffic conditions in Mexico City (MC) is proposed. Data acquisition was carried out over diverse MC routes, representing travel under congested and uncongested conditions, using the chase-car approach. Thirteen different on-road patterns, including the four main access roads to MC, trips in both directions and different timetables, a total of 108 trips spanning 1044 km were evaluated in this study. The MC cycle lasts 1360 seconds with a distance of 8.8 km and average speed of 23.4 km h(-1). Both maximum speed (73.6 km h(-1)) and maximum acceleration (2.22 km h(-1)s(-1)) are lower than those of the new vehicles certification employed in Mexico ,FTP-75 cycle., that is, the MC cycle exhibits less cruising time and more transient events than the FTP cycle. A total of 30 light duty gasoline vehicles were classified into different technological groups and tested in an FTP-75 and MC driving cycles in order to compare their emission factors A potential concern is that in Mexico manufacturers design vehicles to meet the emission standards in the FTP, but emission levels increase significantly in a more representative cycle of present driving patterns in the Metropolitan Area of Mexico City (MAMC). The use of a more representative cycle during certification testing, would provide an incentive for vehicle manufacturers to design emissions control systems to remain effective during operation modes that are not currently represented in the official test procedures used in the certification process. Based on the results of the study, the use of MC cycle, which better represents current day driving patterns during testing of vehicle fleets in emissions laboratories, would improve the accuracy of emissions factors used in the MAMC emissions inventories. PMID- 15791796 TI - Effective diffusion coefficients of glucose in artificial biofilms. AB - A technique using horizontal attenuated total reflection (HATR) by Fourier transform infrared (FTIR) spectrometry was demonstrated for the determination of effective diffusion coefficient (De) of an inert solute in biofilm. Glucose was the selected solute, and agarose (1%) hydrogel films containing various concentrations of activated sludge biomass were used to simulate the biofilm. The agarose films were formed on the surface of an internal reflection crystal in contact with a bulk solution containing 0.5M glucose. Glucose molecules diffused through the film by concentration gradient, and the glucose concentration at the biofilm-crystal interface was measured over time by HATR-FTIR. Based on the glucose concentration and film thickness, the De of glucose was calculated according to the Fick's Law. Results showed that De of glucose in the biomass free agarose films averaged 6.46 +/- 0.21 x 10(6) cm(2) s(-1), which is about 5% lower than the average reported De of glucose in water. The De of glucose decreased with increasing biomass concentration in the artificial biofilm. For the agarose films containing 0.45%, 0.90% and 1.80% of biomass, the De of glucose were lowered to 6.38 +/- 0.22 x 10(-6), 6.08 +/- 0.23 x 10(-6) and 5.62 +/- 0.17 x 10(4) cm(2) s(-1), respectively. PMID- 15791797 TI - Respirometry for assessing the biodegradation of petroleum hydrocarbons. AB - The respiration method using the Micro-Oxymax respirometer was applied to evaluate the bioremediation potential of hydrocarbon-contaminated soils in two biopiles at the oil refinery in Czechowice-Dziedzice, Poland. In biopiles 1 and 2, two different technologies, i.e., enhanced (engineered) bioremediation and monitored natural attenuation (MNA) were used, respectively. In biopiles 1 and 2, the bioremediation process lasted 6 years and 8 months, respectively. The biodegradation of petroleum hydrocarbons was evaluated on the basis of CO2 production and O2 uptake. The CO2 production and O2 consumption rates during hydrocarbon biodegradation were calculated from the slopes of cumulative curve linear regressions. The results confirmed the hydrocarbon biodegradation process in both biopiles. However, in biopile 2 the process was more effective compared to biopile 1. In biopile 2, the O2 consumption and CO2 production means were 3.37 and 2.4 milliliters per kilogram of soil (dry weight) per minute, respectively. Whereas, in biopile 1, the O2 consumption and CO2 production means were 1.52 and 1.07 milliliters per kilogram of soil (dry weight) per minute, respectively. The mean biodegradation rate for biopile 2 was two times higher--67 mg hydrocarbons kg d.w.(-1)day(-1) compared with biopile 1, where the mean was 30 mg hydrocarbons kg d.w.(-1)day(-l). The results were correlated with petroleum hydrocarbon concentrations and microbial activity measured by dehydrogenase assay. PMID- 15791798 TI - Control of trichloroethylene emissions from sparging systems by horizontal bio- and chemo- barriers. AB - The scope of this study was to develop a continuous system to clean-up a trichloroethylene (TCE) contaminated gas stream, where biotic and abiotic removal mechanisms are undertaken sequentially simulating the horizontal bio- and chemo barriers proposed for the in-situ remediation of the contaminated sites. The bio- and chemo-barriers were simulated by using glass columns packed with granular anaerobic mixed culture and Fe(0) filings, respectively. The effect of gas residence time, which is adjusted by the gas flowrate, on the TCE removal efficiency of the reactor system was investigated. TCE removal efficiency of over 90% was achieved at gas residence times above 1hr. Furthermore, the effluent of reactor system contained only ethane and ethylene, which are non-toxic by products of TCE reduction reactions, along with trace amounts of TCE. PMID- 15791799 TI - The effect of nutrient deficiency on removal of organic solvents from textile manufacturing wastewater during activated sludge treatment. AB - Textile manufacturing wastewater is often deficient in nitrogen and phosphorus and contains hazardous solvents, including methyl ethyl ketone (MEK), methyl isobutyl ketone (MIBK), toluene (TOL), and xylenes (XYL). The objectives of this study were to evaluate the effectiveness of a short-term batch assay for predicting when a nutrient deficient condition exists in textile wastewater activated sludge, and to determine if nutrient deficiency affects biodegradation of MEK, MIBK, TOL,and p-XYL to a greater or lesser extent than bulk soluble chemical oxygen demand (sCOD). Addition of N + P significantly improved sCOD removal during treatment of textile wastewater in laboratory-scale sequencing batch reactors (SBRs). Batch tests using mixed liquor suspended solids (MLSS) from the SBRs correctly predicted the nutrient deficiency in the reactors that received unamended wastewater. During batch tests in sealed containers (to prevent volatilization) when N + P were added, the solvents biodegraded faster and to a greater extent than the bulk wastewater sCOD. MEK and MIBK were also completely consumed in MLSS from the SBR that received unamended wastewater, indicating that a shortage of nutrients did not significantly impact biodegradation of these ketones. However, nutrient deficient conditions significantly decreased the rate of TOL and p-XYL biodegradation. The difference in biodegradability of the ketones and monoaromatics under nutrient deficient conditions may be related to loss of plasmids required for aerobic catabolism of TOL and p-XYL. These results demonstrate that N + P addition to nutrient deficient textile wastewater improves bulk sCOD removal and also significantly improves the biodegradability of TOL and p-XYL, thereby reducing the amount released to the atmosphere by volatilization. PMID- 15791800 TI - EDTA leaching of copper from contaminated soils: experimental study and transport model application for parameters estimation. AB - EDTA column's extraction were performed on four artificially contaminated soils, followed by the percolation of pure water to ensure the removal of EDTA entrapped in soil. The investigated soils were characterized by a content of organic matter ranging from 1% to 25%. A mathematical model was developed for copper leaching from the soils. The correlation of experimental data shows that while the EDTA transport was not affected by the soil organic fraction, the simulated transport of Cu-EDTA complexes in solution exhibited a delay effect due to the chemical adsorption of such complexes onto the soil organic matter. The distribution coefficient (Kd), between the solid and liquid phases showed a logarithmic increase when the organic content of soil increased. The column's hydrodynamic characteristics (alphaL) and the kinetic coefficients (k) of chelant extraction were estimated independently for each soil. A correlation of kinetic coefficients data versus soil's organic content was also proposed: experimental data showed, in the investigated range, a logarithmic dependence of k on the organic content of soil. PMID- 15791801 TI - SBR performance with an excessive long sludge age. AB - In the UK water industry, the sequencing batch reactor (SBR) has been seen as a technically-economically advantageous option as an alternative to the continuous flow activated sludge process (ASP). The technology has enjoyed a booming period over the last decade, with many installations now operational. These installations appear to have achieved various degrees of success, but not without operational and performance issues. This paper reviews the typical design practices of SBR, and focuses on one particular SBR supplied by Biobubble Ltd within the spectrum of various SBR technologies. Biobubble SBR is designed as a true SBR with discrete phases of fill, aerate, settle and decant. It often guarantees an effluent quality of 15: 10: 2 (TSS : BOD : AmmN) and a low sludge production. Its main operating parameters are distinct in comparison with continuous flow ASP, and indeed other 'mainstream' SBRs. Differentiation can be seen on plant sizing, sequence control and equipment configuration. Studies have been carried out by Anglian Water Services Ltd to gain a better understanding of this type of SBRs. It has been found that the reliable performance is due to the long hydraulic retention time (HRT) of about 70 hours (aerated), which provide high,tolerance against shock loadings. The good settlement is a result of a long settling phase of 1.5 hours and a low fill time ratio (FTR) of about 0.05, creating a feast/famine condition similar to a plug-flow ASP. The low sludge yield of 0.33 kgSS x kgBOD(-1) is a result of a long sludge retention time (SRT) of about 200 days, which enables aerobic sludge digestion in the SBRs. PMID- 15791802 TI - Influence of lepidocrocite (gamma-FeOOH) on Escherichia coli cultivability in drinking water. AB - A washed suspension of the bacteria Escherichia coli, pre-grown on a complex culture medium, was stored in sterilized drinking water for 21 days at 25 degrees C in glass flasks in order to assess the effect of iron corrosion products on the persistenceof the bacteria in drinking water. Four conditions were tested: aerobic with 50 mM lepidocrocite (gamma-FeOOH, an insoluble iron corrosion product), anaerobic with 50 mM lepidocrocite, aerobic without lepidocrocite and anaerobic without lepidocrocite. The survival of E. coli was monitored by their cultivability and their membrane integrity (propidium iodide staining). When the samples were not supplemented with the iron oxide, the cultivability and cell integrity of the bacteria were dramatically altered: from the 10(7) initially added, only 10 CFU ml(-1) remained after 21 days; 90% of the cells exhibited membrane alteration after 2 weeks of storage. In contrast, bacteria with lepidocrocite preserved their cultivability and integrity over the 21 days of storage. In the presence of di-oxygen and without iron oxide, the alteration of cell cultivability was more pronounced than that in anaerobic conditions, suggesting that oxidative stress was part of the phenomenon. When the cells were pre-grown in a growth medium supplemented by a large excess of an easily available form of iron (ferric-citrate), the cells stored a higher amount of iron and persisted one week longer in the iron-free drinking water than cells pre grown in the standard growth medium. Therefore, in an oligotrophic environment like drinking water, E. coli cells can find the ability to survive a long time through the presence of iron corrosion products. The necessity of controlling the corrosiveness of drinking water for sanitary reasons is therefore emphasized by this study. PMID- 15791803 TI - Phosphate elimination and recovery from wastewater by active filtration using crushed gas concrete. AB - Laboratory long-term upstream fixed-bed experiments were carried out to investigate the efficiency of phosphorus elimination with the effluent of a biological sewage treatment plant using crushed gas concrete. The development of the pH-value in the column outflow as well as the reaction kinetics was investigated. Furthermore, the phosphorus yield was balanced for phosphorus recovery and the calcium phosphate compounds generated were specified by mineralogical analysis methods. These activities were followed by a study with respect to the suitability of the material as raw material for the phosphate industry. PMID- 15791804 TI - Genes: philosophical analyses put to the test. AB - This paper describes one complete and one ongoing empirical study in which philosophical analyses of the concept of the gene were operationalized and tested against questionnaire data obtained from working biologists to determine whether and when biologists conceive genes in the ways suggested. These studies throw light on how different gene concepts contribute to biological research. Their aim is not to arrive at one or more correct 'definitions' of the gene, but rather to map out the variation in the gene concept and to explore its causes and its effects. PMID- 15791805 TI - What concept analysis in philosophy of science should be (and why competing philosophical analyses of gene concepts cannot be tested by polling scientists). AB - What should philosophers of science accomplish when they analyze scientific concepts and interpret scientific knowledge? What is concept analysis if it is not a description of the way scientists actually think? I investigate these questions by using Hans Reichenbach's account of the descriptive, critical, and advisory tasks of philosophy of science to examine Karola Stotz and Paul Griffiths' idea that poll-based methodologies can test philosophical analyses of scientific concepts. Using Reichenbach's account as a point of departure, I argue that philosophy of science should identify and clarify epistemic virtues and describe scientific knowledge in relation to these virtues. The role of concept analysis is to articulate scientific concepts in ways that help reveal epistemic virtues and limitations of particular sciences. This means an analysis of the gene concept(s) should help clarify the explanatory power and limitations of gene based explanations, and should help account for the investigative utility and biases of gene-centered sciences. I argue that a philosophical analysis of gene concept(s) that helps achieve these critical aims should not be rejected on the basis of poll-based studies even if such studies could show that professional biologists don't actually use gene terminology in precise ways corresponding to the philosophical analysis. PMID- 15791807 TI - Alternative splicing, the gene concept, and evolution. AB - Alternative splicing allows for the production of many gene products from a single coding sequence. I introduce the concept of alternative splicing via some examples. I then discuss some current hypotheses about the explanatory role of alternative splicing, including the claim that splicing is a significant contributor to the difference in complexity between the human genome and proteosome. Hypotheses such as these bring into question our working concepts of the gene. I examine several gene concepts introduced to cope with processes such as alternative splicing. Next I introduce some hypotheses about the evolution of mechanisms alternative splicing in higher organisms. I conclude that attention to alternative splicing reveals that we adopt an attitude that developmental theorizing must inform evolutionary theorizing and vice versa. PMID- 15791806 TI - Molecular epigenesis, molecular pleiotropy, and molecular gene definitions. AB - Recent work on gene concepts has been influenced by recognition of the extent to which RNA transcripts from a given DNA sequence yield different products in different cellular environments. These transcripts are altered in many ways and yield many products based, somehow, on the sequence of nucleotides in the DNA. I focus on alternative splicing of RNA transcripts (which often yields distinct proteins from the same raw transcript) and on 'gene sharing', in which a single gene produces distinct proteins with the exact same amino acid sequence. These are instances of molecular pleiotropy, in which distinct molecules are derived from a single putative gene. In such cases the cellular and external environments play major roles in determining which protein is produced. Where there is molecular pleiotropy, alternative gene concepts are naturally deployed; molecular epigenesis (revision of sequence-based information by altering molecular conformations or by action of non-informational molecules) plays a major role in orderly development. These results show that gene concepts in molecular biology do, and should, have both structural and functional components. They also show the need for a plurality of gene concepts and reveal fundamental difficulties in stabilizing gene concepts solely by reference to nucleotide sequence. PMID- 15791808 TI - Long live the genome! So should the gene. AB - Developments in the sequencing of whole genomes and in simultaneously surveying many thousands of transcription and translation products of specific cells have ushered in a conceptual revolution in genetics that rationally introduces top down, holistic analyses. This emphasized the futility of attempts to reduce genes to structurally discrete entities along the genome, and the need to return to Johannsen's definition of a gene as 'something' that refers to an invariant entity of inheritance and development. We may view genes either as generic terms for units of inheritance whose referents are pragmatic ad hoc and context dependent, or as (epistemologically) representing entities of cell functions. It is cellular functions that determine the structural referents along the DNA. Structures that happened to secure specific functions that were essential for or conducive to the survival of cells were selected for. With natural selection being the etiological background of genes as functions, genes obtain again their theoretical role as intervening variables, abstractive variables that purely 'summarize' characters. The importance of DNA sequences is that of all possible phenotypes these are the most basic ones from which we can read off the genotype directly. PMID- 15791809 TI - [Cancer at the esophagogastric junction]. AB - The definition of cancer at the esophagogastric junction has been discussed for over 30 years. Although at present, two classifications advocated by Nishi or Siewert have been used, these classifications are still controversial from the viewpoints of anatomy, physiology, histology, diagnosis and treatment. We should analyze a large number of patients based on the same definition of cancer at the esophagogastric junction. Adjustment of the Japanese rules for cancer of the esophagus and stomach is necessary, and a treatment strategy should be established for cancer at the esophagogastric junction. PMID- 15791810 TI - [Myelodysplastic syndrome]. AB - Myelodysplastic syndrome (MDS) is a heterogenous syndrome which has features of refractory anemia and preleukemic state due to the ineffective hematopoiesis of bone marrow cells. MDS is classified by the FAB classification and/or WHO classification, and has an International Prognostic Scoring System (IPSS) for evaluation of the prognosis. Available therapies consist of a variety of modalities from supportive therapy to hematopoietic stem cell transplantation, and are selected according to the condition of each patient. PMID- 15791811 TI - [Current and new therapeutic strategies in acute myeloid leukemia]. AB - Seventy to 80% of patients with acute myeloid leukemia (AML) achieve complete remission (CR) by chemotherapy, but more than 50% of them then relapse. Phase III clinical trials in the treatment of patients with previously untreated AML and acute promyelocytic leukemia (APL) are ongoing in Japan (JALSG AML 201, APL 204). And continuous efforts are being made to improve the efficacy of chemotherapy. We discussed six topics in the treatment of AML. (1) To determine whether adding the MDR-1 modulator to chemotherapy provided clinical benefits to patients with AML and high-risk myelodysplastic syndrome (MDS), a phase III randomized study was performed using PSC 833. CR rates and overall survival (OS) were not improved by using PSC 833 compared to chemotherapy alone. (2) A large randomized study selectively focused on the G-CSF priming was performed. Among patients in this study attaining CR, the probability of relapse was reduced when they had been assigned to treatment with G-CSF along with induction chemotherapy. The benefit of chemotherapy-sensitization by G-CSF was particularly evident among the intermediate-risk. (3) Fludarabine in addition to Ara-C increases the accumulation of Ara-CTP, which is responsible for the cytotoxic effect in leukemic blasts. In a randomized phase III trial, patients with high-risk MDS or patients with AML were randomized to receive 2 induction courses consisting of Ara-C and G-CSF during and after chemotherapy with or without fludarabine (FLAG versus AG). Although Ara-CTP accumulation in leukemic cells after FLAG was enhanced, the clinical outcome in terms of CR rate, OS, event-free survival, and disease-free survival was not significantly improved by combining fludarabine with Ara-C. (4) Calicheamicin-conjugated humanized anti-CD 33 mouse monoclonal antibody, mylotarg, has recently been introduced. In combined phase II studies of 277 patients with CD 33-positive AML in their first relapse, the overall response rate was 26%. (5) Arsenic trioxide (ATO) has been established as a highly effective therapy for patients with APL, even for those with disease refractory to ATRA. ATO was recently approved in Japan. (6) There has been great interest in developing FLT 3 inhibitors because of the high frequency and poor prognosis of AML patients with mutant FLT 3. Some compounds are currently under development. PMID- 15791812 TI - [Imatinib therapy for patients with chronic myelogenous leukemia]. AB - Chronic myelogenous leukemia (CML) is a clonal hematopoietic disorder caused by the reciprocal translocation between chromosome 9 and 22. As a result of this translocation, a novel fusion gene, BCR-ABL, is created on Philadelphia (Ph) chromosome, and the constitutive activity of the BCR-ABL protein tyrosine kinase plays a critical role in the disease pathogenesis. Imatinib mesylate, a selective BCR-ABL tyrosine kinase inhibitor, was first given to a patient with CML in June 1998. Since then, it has continued to demonstrate remarkable efficacy in treating patients with CML. Based upon the results of early phase I and II studies, a phase III study (IRIS Study) that was randomized to first-line imatinib (400 mg/day) or to standard treatment with interferon+low-dose Ara-C, was conducted on 1,106 patients newly diagnosed (within 6 months) with chronic-phase CML. After median follow-up of 30 months, imatinib showed significantly superior tolerability, hematologic and cytogenetic responses (major cytogenetic response, 90%; complete cytogenetic response, 82%), and overall survival (95% without censoring allo-HSCT). Although imatinib is the first-line therapy and has changed the paradigm of CML treatment strategy, questions remain as to the meaning of cytogenetic and molecular response, curability, optimal dose, and relation with allo-HSCT. PMID- 15791814 TI - [Progress in therapeutic strategy for malignant lymphoma]. AB - Dramatic progress in therapeutic strategy for malignant lymphoma has been made during recent few years because of the development of new drugs or new therapeutic modalities such as rituximab, purine analogue, purged autologous PBSCT or allogeneic PBSCT by the reduced intensity technique. Rituximab in particular changed the golden standard therapy for previously untreated patients with diffuse large B-cell lymphoma from CHOP therapy to rituximab-CHOP (R-CHOP) therapy in all risk groups. In follicular lymphoma with no treatment strategies associated with curative potential and median survival in the range of 8 to 13 years, prolonged progression-free survival has been reported by maintenance-use of rituximab, R-CHOP therapy, purine analog, in vivo purged auto-PBSCT by rituximab, or reduced intensity stem-cell transplantation (RIST), although no curable survival benefit has yet been demonstrated by any strategies. Short courses (4 courses) of ABVD followed by involved field irradiation therapy (IFRT) for localized early-stage Hodgkin lymphoma (HL), and full courses (6-8 courses) of ABVD (d) for advanced stage HL are the golden standard therapy for HL, respectively. Clinical trials of new therapies with more efficacy and less toxicity have been undertaken. PMID- 15791813 TI - [Multiple myeloma--recent advances in diagnosis and treatment]. AB - Multiple myeloma (MM) is a systemic malignancy of pathologic plasma cells that is treatable with various chemotherapeutic agents and irradiation, but rarely curable. The mean age of affected patients is mid-60s. Recent clinical studies have confirmed that high-dose chemotherapy with stem cell transplantation is the standard therapy for MM. Furthermore, novel therapeutic agents, such as thalidomide and other immunomodulatory agents have been extensively investigated. Recent advances in the diagnosis and treatment were reviewed and summarized. PMID- 15791815 TI - [Evaluation of pre-operative administration of TS-1 in patients with advanced gastric or colorectal cancer--estimation of pathological effectiveness for postoperative adjuvant chemotherapy with TS-1]. AB - We performed surgical resections in 6 cases of advanced gastric cancer and 4 cases of colorectal cancer after preoperatively treating them with TS-1 at a daily dose of 80-100 mg/body for 2 weeks, and evaluated whether one can estimate their sensitivity to TS-1 by a pathological examination. Case 1 of type 3 advanced gastric cancer underwent surgery after one week interval following oral administration of TS-1 at a daily dose of 80 mg/body for 2 weeks. Surprisingly, the pathological examination revealed complete disappearance of cancer cells in the resected stomach and no cancer cells in the regional lymphnodes, judged grade 3 in pathological effectiveness. Case 2 of type 2 advanced gastric cancer was treated with TS-1 at a daily dose of 100 mg/body for 2 weeks and underwent surgery after a three-week interval due to the complication of pneumonia. The pathological effectiveness was judged grade 2 in the resected stomach, and no cancer cells were detected in the regional lymphnodes. In both cases, the postoperative course was uneventful, and no adverse effects were detected. In these cases, their high sensitivity to TS-1 was clearly confirmed, and now they have been treated with TS-1 for the postoperative adjuvant chemotherapy, and have undergone regular check-ups at our outpatient clinic in good condition. Recently, we performed the same protocol in 6 cases of advanced gastric cancer including these 2 cases and also in 4 cases of advanced colorectal cancer. This protocol was found useful for evaluating the pathological effect by TS-1. We consider the protocol quite useful and helpful in determining a suitable regimen for postoperative adjuvant chemotherapy. PMID- 15791816 TI - [Study of three-weekly docetaxel and weekly trastuzumab treatment in HER 2 overexpressing metastatic breast cancer patients]. AB - Docetaxel and trastuzumab can be considered to be active drugs for HER 2 overexpressing metastatic breast cancer (MBC). This study was conducted to determine the activity of combination therapy with docetaxel and trastuzumab in MBC patients (pts) by assessing the response rate (RR), time to progression (TTP) and safety. We administered the combination of docetaxel 70 mg/m2 every 3 weeks and trastuzumab using a 4 mg/kg loading dose and thereafter 2 mg/kg weekly. One cycle was three weeks. Between March 2002 and May 2003, 40 pts with HER 2 positive (3+by immunohistochemistry 39, FISH+1) MBC were enrolled in this study, and 39 pts proved eligible. The overall RR was 72% (28/39) [95%CI 55.1%-85.0%], with 6 CR, 22 PR, 7 SD, 1 PD and 3 NE. The median follow-up time was 14.3 months, while the TTP was 6.5 months (range, 0.6-19.8), median OS has not yet been reached. The number of pts assessable for safety was 40. Hematological Grade 3-4 toxicities were leukopenia 87.5% (35/40) and neutropenia 82.5% (33/40). Non hematological Grade 3 toxicities were weight gain in 2 pts, and anorexia, neuropathy, fever and rash in one pt each. The combination of docetaxel and trastuzumab was a well-tolerated and very active regimen for the treatment of pts with HER 2-overexpressing MBC. PMID- 15791817 TI - [Intrathoracic infusion with a combination of low-dose minocycline, OK-432 and cisplatin for malignant pleural effusion]. AB - We investigated the effectiveness and complications of intrathoracic infusion with a combination of cisplatin, OK-432, and minocycline for malignant pleural effusion. All patients were hospitalized with chest tube drainage of pleural effusion until the daily drainage volume was less than 100 ml. Twenty-five mg of minocycline, 1 to 3 KE of OK-432, and 5 to 10 mg of cisplatin were instilled into the pleural space. The administration was repeated until drainage effusion disappeared. Therapeutic effect was evaluated according to the following criteria: (1) excellent, no fluid reaccumulation for at least 4 weeks as determined by chest radiogram and clinical evaluation; (2) effective, fluid reaccumulation less than 50% of original effusion with no need of thoracentesis for symptomatic relief within 4 weeks after treatment; and (3) failure, reaccumulation of more than 50% of the original effusion requiring thoracentesis to relieve symptoms within 4 weeks of treatment. Twelve patients with malignant effusion received the combination treatment; 11 patients had primary lung cancer and one had metastatic lung tumor from cancer of the rectum. In all cases, the histology or cytology revealed adenocarcinoma. Eleven of the 12 patients had an excellent response with relief of clinical symptoms. The remaining case failed to show any improvement. Complications such as local pain, fever, nausea, and vomiting were mild and transient. We conclude that combination administration of low-dose minocycline, OK-432, and cisplatin into the thoracic cavity for malignant effusion is an effective alternative treatment with the potential for improvement of the general condition and reduced morbidity. PMID- 15791818 TI - Cost-effectiveness of letrozole versus tamoxifen as first-line hormonal therapy in treating postmenopausal women with advanced breast cancer in Japan. AB - The objective of this study is to evaluate the cost-effectiveness of letrozole compared with tamoxifen as first-line therapy in post-menopausal women with advanced breast cancer in Japan. A Markov analytical model was developed to estimate life-year (LY) expectancies, using key transition probabilities obtained from the results of a multinational phase III trial, a literature review and a Japanese medical expert panel. Direct medical costs were estimated, from the payer's perspective, using the expected resource utilization provided by the expert panel, the medical fee table and drug tariff under the national health insurance system. The expected overall life-years (LYs) obtained were 3.68 years for letrozole arm and 3.09 years for tamoxifen arm, showing incremental LYs of 0.59 years in patients receiving letrozole. The total expected costs were 3,644,588 yen (33,133 US dollars) for letrozole arm and 3,322,111 yen (30,201 US dollars) for tamoxifen arm, resulting in a mean incremental cost-effectiveness ratio (ICER) of 546,571 yen (4,969 US dollars) per life-year gained, while the 5 th percentile of ICER showed letrozole dominating tamoxifen and the 95th percentile was 2,310,593 yen (21,005 US dollars). The results suggest that letrozole is a clinically beneficial and cost-effective treatment option when compared with tamoxifen in first-line therapy for advanced breast cancer in Japan. PMID- 15791819 TI - [Weekly paclitaxel administration and intraabdominal CBDCA injection possibly beneficial treatment for recurrent breast cancer associated with metastatic ovarian cancer and peritoneal dissemination after operation--a case report]. AB - A 46-year-old woman who had received mastectomy for breast cancer 6 years earlier complained of abdominal distension. Computed tomography and ultrasonography revealed massive ascites and ovarian swelling of both sides. She was diagnosed as having primary ovarian cancer and peritoneal dissemination, and underwent a total hysterectomy as well as ovarectomy on both sides. After surgery, she received a sequential chemotherapy, ie, intraabdominal injection of carboplatin (CBDCA 300 mg/m2) and div administration of paclitaxel (PTX 180 mg/m2) as a standard regimen for advanced ovarian cancer. However, detailed histological examinations showed that the ovarian cancer had metastasized from her breast cancer. It is well-known that breast cancer easily metastasizes to the bone, liver, pleura and lymph node, but rarely to the ovarium or peritoneum when chemotherapy is conducted. Therefore, no standard therapy has been established for breast cancer metastasizing to the ovarium. Our patient received 4 cycles of weekly administration of PTX (PTX 80 mg/m2, 3 consecutive weeks, 1-week break), followed by oral administration of doxifluridine+anastrozole on an outpatient basis. No evidence of recurrence of breast cancer has been noted 1 year after surgery. This result suggests that weekly administration of PTX and intraabdominal injection of CBDCA might be beneficial in the treatment of recurrent breast cancer associated with metastatic ovarian cancer and peritoneal dissemination after operation. PMID- 15791820 TI - [Effective reduction of elevated tumor markers by exemestane--a 12-year follow-up of a case of bone metastatic breast cancer]. AB - A 75-year-old woman with low back pain had attended a local orthopedic clinic since around December 1990, but no remission had been achieved. The patient also had a gait disturbance, and visited our hospital for detailed examinations in May 1992. Bone scintigraphy revealed metastatic tumors of L 1 and L 2. Histopathological findings and tumor marker measurements led to a diagnosis of primary breast cancer. The patient was treated with TAM (20 mg/day) and 5-FU (150 mg/day). The tumor marker levels showed repeated cycles of slight increases and decreases. Acute elevations of the tumor markers occurring on January 24, 1997, and January 5, 2001, were successfully treated with MPA (800 mg/day) and 5'-DFUR (600 mg/day), but continuous administration of these drugs was difficult because of their adverse effects. A significant increase in the tumor marker levels (CA15 3 600 U/ml, CEA 197 ng/ml) was again observed on December 2, 2003. The patient showed no favorable response to the combination of MPA and 5'-DFUR but had persistent back pain. Exemestane given at 25 mg/day markedly improved both clinical symptoms and tumor marker levels. The results indicate that exemestane would be an effective hormone therapy for metastatic breast cancer. PMID- 15791821 TI - [A case of thymic carcinoma responding to combination chemotherapy with nedaplatin, etoposide, and ifosfamide]. AB - We described a case of thymic carcinoma that responded remarkably to combined chemotherapy with etoposide (ETP), ifosfamide (IFO) and nedaplatin. A 68-year-old woman was admitted to our hospital because of hoarseness and dysphasia. Chest computed tomographic (CT) scans showed an anterior mediastinal tumor. Using CT guided needle biopsy, the diagnosis was squamous cell type of thymic carcinoma. We gave her modified VIP therapy with ETP, IFO and nedaplatin. The tumor contraction response was evident after 3 courses of treatment. This case suggested that nedaplatin may be one of the promising agents for thymic carcinoma chemotherapy. PMID- 15791822 TI - [Evaluation of low-dose FP therapy as adjuvant therapy for gastric cancer]. AB - In the present study, we demonstrate the result of low-dose FP treatment as an adjuvant therapy for 30 patients of stage II or more progressive gastric cancer. 5-FU and CDDP were injected intravenously for 10 days from day 1 through day 5, and day 8 through 12 for 2 weeks at a dose of 250 mg/body and 10 mg/body, or for 14 days at a dose of 250 mg/m2/day and 5 mg/m2/day, respectively. Patients were excluded if they received less than 80% of the respective doses in a course of treatment by the protocol. This constituted a course of chemotherapy, which was repeated every 4 weeks. Grade 3 neutropenia was observed in one case. Other toxicities were anorexia, nausea, weight loss, diarrhea, general fatigue and elevation of serum creatinine, but they were not so severe. The two-year survival rate was 100% in cur A cases, 85% in cur B, and 0% in cur C. The median survival time of the cur C patients was 10 months. These results indicate that low-dose FP therapy is safe and recommendable for cur A and cur B patients. However, other treatment methods such as sequential chemotherapy are needed for cur C gastric cancer patients. PMID- 15791823 TI - [A case of non-curatively resected scirrhous gastric cancer successfully treated over 2 years with weekly administration of paclitaxel]. AB - We report a patient with far-advanced gastric cancer treated by weekly administration of paclitaxel (TXL) over 2 years. The patient was a 66-year-old female with peritoneal metastasis and remarkable lymph node metastasis of scirrhous gastric cancer. She underwent a non-curative resection with total gastrectomy and splenectomy in May 2002. Postoperative chemotherapy with TS-1 (80 mg/body) was performed. Due to grade 4 neutropenia and grade 2 anorexia, this treatment could not be continued. Three months after surgery, the tumor marker (CA19-9) had elevated to an abnormal level. Alternatively, TXL was administered at a weekly dose of 70 mg/m2 for 3 weeks followed by 6 weeks rest from September 2002. The tumor marker (CA19-9) gradually decreased to the normal level. Because of the long rest interval, 10 courses of treatment could be continued, and the patient has been alive over 2 years with the cancer controlled. There have been few effective chemotherapies for gastric cancer with peritoneal metastasis. Weekly paclitaxel therapy is considered to be effective for the treatment of advanced scirrhous gastric cancer with peritoneal metastasis. PMID- 15791824 TI - [Three cases of multiple colorectal liver metastases responding to hepatic arterial infusion chemotherapy undergoing liver resection]. AB - We encountered three cases of multiple liver metastases of colorectal cancer treated with hepatic resection after hepatic arterial infusion. Case 1: A 55-year old female underwent sigmoidectomy for sigmoid colon cancer and six liver metastases. After responding postoperatively to hepatic arterial infusion chemotherapy (HIA), she underwent resection of liver metastases. Case 2: A 66 year-old man underwent colectomy and hepatic resection for cecal colon cancer and liver metastasis. Multiple liver metastases appeared six months after the operation. HIA resulted in a complete response. Liver metastases recurred twice and liver resection was performed each time. Case 3: A 52-year-old female underwent partial resection of colon and liver for ascending colon cancer and multiple liver metastases. After she responded postoperatively to HIA and systemic chemotherapy, the patient underwent resection of liver metastases. Case 1 has lung metastases at this writing. The other two patients are alive and well, and have been free from recurrence as of 4 years after operation. PMID- 15791825 TI - [A case of bilateral multiple liver metastases with distant lymph node metastases due to rectal cancer successfully treated with hepatic arterial infusion (HAI) of levofolinate (l-LV)/5-fluorouracil (5-FU) and radiotherapy]. AB - The patient was a 43-year-old male with bilateral multiple liver metastases, who had undergone high anterior resection for rectal cancer (ss, n 0, P 0, H 3, M ( ), stage IV). Hepatic arterial infusion (HAI) of low-dose CDDP (10 mg/body) and 5 FU (250 mg/body), 5 times a week, was ineffective for the liver metastases. Consequently, HAI of levofolinate (425 mg/body) and 5-FU (1,000 mg/body), once a week, was attempted. All metastatic liver tumors diminished apparently with calcification after the treatment (PR). Tumor marker (CA19-9 and CEA) levels decreased to less than one-tenth of the pretreatment levels and stabilized for approximately seven months. Mediastinal lymph node metastases, paraaortic lymph node metastases and tumor thrombus in the inferior vena cava were successfully treated with systemic chemotherapy using levofolinate and 5-FU and/or radiotherapy. Although the liver and lung metastases showed rapid growth, the patient died 2 years after the diagnosis of liver metastases. The liver metastases were well controlled for about 20 months. It is important to select interdisciplinary therapies according to the site of the metastases due to rectal cancer. PMID- 15791826 TI - [Induction of CPT-11 in a patient on hemodialysis with metastatic rectal cancer]. AB - We report a case on hemodialysis with metastatic rectal cancer who was introduced to CPT-11. Although the expected pharmacokinetics was shown 24 hours post dialysis with the infusion of dose-reduced CPT-11, grade 4 neutropenia was observed. Considering the chronic renal failure status with latent lower function of multiple organs, the dose escalation method was recommended while watching the pharmacokinetics. CPT-11 is not only the key compound for metastatic colorectal cancer, but is also effective with several other cancers. It is important for cancer patients with chronic renal failure that the feasibility and efficacy of CPT-11 should be determined by future study. PMID- 15791827 TI - [Effect of steroid on antiemetic for side effect of anticancer chemotherapy]. AB - The side effect of anticancer agents such as nausea and vomiting frequently interrupt chemotherapy. To reduce these side effects, 5-hydroxytryptamine (5-HT3) receptor antagonist or metoclopramide is administered combined with steroid. In this study, we examined the effect of 5-HT3 receptor antagonist on the frequency of nausea and vomiting in a male cancer patient treated with/without steroid. This patient in his sixties had esophageal cancer (stage IV). He was administered nedaplatin 100 mg/day for 1 day and then 5-fluorouracil (5-FU) 750 mg/day for 5 days combined with radiotherapy (60 Gy) as one cycle of this chemotherapy. In the first cycle, 5-HT3 receptor antagonist was administered, and in the second, the antagonist was administered after treatment with steroid. The blood levels of total bilirubin, GOT, GPT, BUN, Cre, Na, K and Cl were stable normally during both cycles of the chemotherapy, indicating that the hepatopathy and nephropathy which cause nausea and vomiting did not occur in these periods. The frequency and period of the nausea and vomiting were one-third decreased, respectively, by the combination of 5-HT3 receptor antagonist and steroid. PMID- 15791828 TI - [Limited surgery for early gastric cancer using lymphatic basin dissection--a sure method of sentinel node biopsy for gastric cancer]. AB - The results of our sentinel node biopsy for early-stage gastric cancer had a high predictive value for nodal metastasis, with a sensitivity of 85% (34/40) and an accuracy of 98% (259/265). Therefore, sentinel node biopsy is expected to be decided as an adaptation of limited surgery for early gastric cancer. However, there were two serious problems in sentinel node biopsy for gastric cancer; one was the great difficulty of sentinel node detection and biopsy in the operative field, and the other was the possibility of false negative cases in the frozen section diagnosis. To solve these problems, we developed lymphatic basin dissection, a new technique suitable for sentinel node biopsy of gastric cancer. The lymphatic basin is an own lymphatic components dyed in blue in the dye method. In the lymphatic basin dissection method, sentinel nodes are detected and harvested at the back table after en bloc dissection of the lymphatic basins. Lymphatic basin dissection is an excellent method because of the certainty in sentinel node biopsy, and the sure back-up dissection. Lymphatic basin dissection and following limited surgery was performed on 143 patients for early-stage gastric cancer in our hospital. No patient had a recurrence of gastric cancer; 9 patients died of other diseases, and the other 134 survived. PMID- 15791829 TI - [Diagnostic significance of tumor markers for gynecologic malignancies]. AB - Gynecologic malignancies include ovarian cancer, uterine cervical cancer, endometrial cancer, and trophoblastic neoplasms. With ovarian tumors, due to their location within the abdominal cavity, it is difficult to make a preoperative pathological diagnosis of cancer without laparotomy. From this point of view, the use of tumor markers that consist of carbohydrate antigens, such as CA 125, in addition to diagnostic imaging are useful in the diagnosis of ovarian cancer. SCC antigen, a marker for squamous cell carcinoma, is clinically useful in the management of advanced cervical cancer. At present, there are no useful tumor markers for endometrial cancer that exhibit both high sensitivity and specificity, although CA 125 is often used in clinical practice. Finally, human chorionic gonadotropin (hCG) serves as an ideal tumor marker for trophoblastic disease; however, the incidence of trophoblastic neoplasms has decreased dramatically with the incorporation of strict clinical management of post-molar disease as well as with the overall decrease in the number of pregnancies. PMID- 15791830 TI - Biographical sketch of academician Qiu Fazu. PMID- 15791831 TI - Investigation on the effects of soluble programmed death-1 (sPD-1) enhancing anti tumor immune response. AB - By using semi-quantitative RT-PCR method, it was found that PD-L1 mRNA but not PD L2 mRNA was expressed in H22 hepatoma cells and both PD-L1 and PD-L2 mRNAs were expressed in tumor tissues of tumor-bearing mice and upregulated as compared with muscle tissues in normal mice and H22 hepatoma cells. PD-L1 and PD-L2 were also expressed on the surface of the activated T cells. The soluble recombinant sPD-1 expressed from the constructed eukaryotic expression vector could enhance the lysis of tumor cells by lymphocytes stimulated specifically with antigen. The expresssion of sPD-1 by local gene therapy on the inoculation site of H22 hepatoma cells could inhibit the growth of tumor. The results of this study indicate that expression of soluble receptor of negative costimulatory molecules could reduce the inhibitory effect on T cells in tumor microenvironment and enhance the cytotoxicity of T cells on tumor cells. This possibly provides a new method of improving efficacy of tumor gene therapy. PMID- 15791832 TI - Prokaryotic expression and preparation of polyantibody of human histydyl-tRNA synthetase related gene. AB - The aim of this study was to express and purify human histydyl-tRNA synthetase related gene and to prepare its polyantibody. The open reading frame was amplified by PCR, and then recombined into prokaryotic expression vector pQE30 and transformed into E. coli M15 for expression. The expressed products were induced by IPTG after the reconstructed pQE30 was transferred into M15. After purified by Ni affinity chromatography, the product was identified to be a single band by SDS-PAGE. The rabbits were inoculated with purified products. High-titer polyantibody was successfully prepared. Highly-purified expression product and prepared polyantibody may provide a good basis for further study. PMID- 15791833 TI - Carboxyl terminus truncated human papillomavirus type 58 L1 protein maintains its bioactivity and ability to form virus-like particles. AB - To prepare carboxyl terminus truncated human papillomavirus type 58 L1 (HPV58L1) protein and evaluate its ability to form virus-like particles, the baculovirus and Sf-9 insect cells was used to express HPV58L1 protein, and pFastBac-Htb containing HPV58L1 gene sequence of carboxyl terminus truncation was generated. Then Sf-9 cells were infected with recombinant baculovirus. After being cultured, the post-infected cells expressing--HPV58L1 protein were harvested and analyzed by SDS-PAGE and Western blot. The ProBond purification system was used for protein purification. The bio-activity of purified protein was identified by mouse erythrocyte hemagglutination assay, and the VLP formation was examined with transmission electron microscope. Our results showed that the recombinant baculovirus was generated and the Sf-9 cells was infected with the recombinant baculovirus, and after collecting, total cellular proteins were extracted. Truncated HPV58L1 protein with MW 58KD was revealed by SDS-PAGE and confirmed by Western blot. The purified L1 proteins under native condition could cause mouse erythrocytes to agglutinate and form VLP. It is concluded that HPV58L1 protein with carboxyl terminus truncation could be efficiently expressed. In baculovirus Sf-9 cells expression system, the purified protein could self-assemble into virions in vitro, and induce agglutination of mouse erythrocytes, indicating that carboxyl terminus truncation does not interfere with the bioactivity of HPV58L1 protein. PMID- 15791834 TI - Effect of interleukin-1beta on the variation of adenylyl cyclase expression in rats with seizures induced by L-glutamate. AB - To explore the mechanism of interleukin-1beta (IL-1beta) in the onset of seizure and the effect of IL-1beta on the expression of adenylyl cyclase (AC) in rats with seizure induced by L-glutamate. Experimental rats were first injected with IL-1beta and then L-glutamate (a dose under the threshold) was injected into the right lateral ventricle. The rats were sacrificed 4 h after the onset of epileptic activity and examined for changes in behavior, immunohistochemistry and compared with those with seizure induced by L-glutamate alone. It was found that the expression of AC in hippocampal and neocortex of rats with seizure induced by IL-1beta and L-glutamate were stronger than that of control group (P<0.05), without significant difference found between the L-glutamate group and IL-1beta plus L-glutamate group in the expression of AC, the latent period and the severity of seizure. When IL-ra were given (i.c.v.) first, there was no epileptic activity and the expression of AC did not increase. There were no differences in the expression of AC of rats with IL-1ra and that of control rats. But when 2 methyl-2-(carboxycyclopropyl) glycine (MCCG) was given (i.c.v.) first, the strongest expression of AC, the shortest latent period and the the most serious seizure activities were observed. The results indicated that IL-1beta could facilitate the onset of epilepsy induced by L-glutamate through IL-1R, metabotropic glutamate receptors might work with IL-1R and the increased expression of AC might be involved in the process. PMID- 15791835 TI - Experimental study of effect of corticosterone on primary cultured hippocampal neurons and their Ca2+/CaMKII expression. AB - To explore the effect of different concentrations of corticosterone (CORT) on primary cultured hippocampal neurons and their Ca2+/CaMK II expression and possible mechanism, the changes of hippocampal neurons were observed in terms of morphology, activity of cells, cell death, concentrations of cytosolic free calcium, and the expression of CaMK II by using MTT assay, flow cytometry, fluorescent labeling of Fura-2/AM and Western blotting after 10(-7), 10(-6) and 10(-5) mol/L of CORT was added to culture medium, The evident effect of 10(-6) and 10(-5) mol/L of CORT on the morphology of hippocampal neuron was found. Compared with control neurons, the activity of the cells was markedly decreased and [Ca2+]i increased in the neurons treated with 10(-6) and 10(-5) mol/L of CORT, but no change was observed in the neuron treated with 10(-7) mol/L of CORT. The death was either by way of apoptosis or necrosis in the cells treated with 10(-6) and 10(-5) mol/L of CORT respectively. The correlation analysis showed that a reverse correlation existed between [Ca2+]i and the expression of CaMK II. Either apoptosis or necrosis occurs in the hippocampal neurons treated with CORT. The increased hippocampal [Ca2+]i is both the result of CORT impairing the hippocampal neurons and the cause of the apoptosis of hippocampal neurons and the decreased CaMK II expression. PMID- 15791836 TI - The comparative study on ultrastructure and immunohistochemistry in AFP negative and positive hepatocellular carcinoma. AB - SUMMARY: To comparatively investigate ultrastructural characteristics and expressions of AFP (alpha-fetoprotein) and Tn (Thomsen-Friedenreich-related antigen) protein in AFP negative (AFP-) and AFP positive (AFP+) primary hepatocellular carcinoma. Fourty-three cases of AFP- and AFP+ hepatocellular carcinoma (HCC) tissues and five cases of normal liver tissues were divided into three groups: control group (normal liver tissue, n=5); AFP+ HCC group (the serum AFP level was higher than 10 ng/ml, n = 22); AFP- HCC group (the serum AFP level was lower than 10 ng/ml, n=21). The ultrastructural morphology was studied by transmission electron microscopy, the expressions of AFP and Tn protein were detected by immunohistochemistry and cell image analysis. 1. The immunohistochemical study showed that (1) the expression intensity and positive rate of Tn protein in AFP- HCC group were markedly higher than that in AFP+ HCC group (P<0.01); (2) The expression intensity of AFP in AFP- HCC group was lower than that in AFP+ HCC group (P<0.01). 2. The transmission electron microscopy demonstrated that some AFP- HCC cells linked closely with each other, others dispersed loosely just as cultured cells, the remarkable morphologic features in AFP- HCC cells were simple organelles, but they were abundant in the free polyribosomes. In AFP+ HCC group, all the HCC cells linked closely together and were rich organelles in their cytoplasm, especially the rough endoplasmic reticula. In addition, mitochondria and Golgi complex were obviously observed. (1) The AFP and Tn protein had discrepancy distribution in AFP- and AFP+ HCC tissues, Tn protein may be one of the early diagnostic indicators in AFP- HCC; (2) The synthetic locations of the AFP and Tn protein were different in hepatocarcinoma cells by ultrastructural observation. PMID- 15791837 TI - Effects of 3,4-dihydroxyacetophenone on cytosolic calcium in pulmonary artery endothelial and smooth muscle cells during acute hypoxia. AB - The effects of 3, 4-Dihydroxyacetophenone (3, 4-DHAP) on cytosolic free calcium [Ca2+]i in pulmonary artery endothelia (PAECs) and smooth muscle cells (PASMCs) during acute hypoxia were studied. Porcine pulmonary artery endothelial and smooth muscle cells (PASMCs) were cultured primarily, and they were divided into 4 groups: groups incubated under normoxia or hypoxia and those with or without treatment with 3,4-DHAP. The [Ca2+]i of both PAECs and PASMCs was measured by determining the fluorescence of fura 2 AM on spetrofluorometer. Our results showed that hypoxia caused significant elevation of [Ca2+]i, in both PAECs and PASMCs, 3,4-DHAP could attenuate the hypoxic elevation of [Ca2+]i only in PASMCs but not in PAECs. It is concluded that 3,4-DHAP decreases the hypoxic elevation of [Ca2+]i in PASMCs. This might contribute to its inhibitory effect on hypoxic pulmonary vasoconstriction. PMID- 15791838 TI - The application of surface electromyography in the assessment of ergonomic risk factors associated with manual lifting tasks. AB - The purpose of this study was to evaluate the ergonomic risk factors associated with manual lifting tasks using surface electromyography (EMG). 13 volunteers lifted loads of 6 and 13 kg at two speeds and at two horizontal distances in 3 different postures and three boxes of different sizes, from floor to knuckle height, performing 72 lifting tasks. For each lift, the surface electromyography signals from the erector spinae muscles, bilaterally at T10 and L3, was recorded. The ergonomic risk factors associated with manual lifting tasks were evaluated by comparing the average amplitude of EMG signals from the erector spinae muscles. The EMG average amplitude for lifting the load of 13 kg was 14.3% greater than that for lifting the load of 6 kg (t=-10.93, P<0.01). The EMG average amplitude at the site of L3 was 10.3% greater than that at the site of T10 (t=-7.98, P<0.01). The EMG average amplitude when performing "fast" lift was 5.9% greater than the "slow" lift (t=-4.63, P<0.01). The posture of lifting affected the EMG average amplitude. It was lowest with semi-squat posture and greatest with squat posture (F=27.76, P<0.01). The result of multiple stepwise regression analysis showed that the loads of lifting, the size of box, horizontal distance, posture of lifting, the site of the spine subjected to force, lifting speed were the factors affecting the EMG average amplitude. The most significant factor was the loads of lifting, followed by the site of the spine subjected to force and the lifting speed in terms of risk. The ergonomic risk factors associated with manual lifting tasks includes the loads, posture, lifting speed, horizontal distance, the site of the spine subjected to force etc. The results of signal amplitude of EMG from the erector spinae muscles showed that semi-squat posture is the best posture for lifting tasks. PMID- 15791839 TI - Effects of rAAV-CD151 and rAAV-antiCD151 on the migration of human tongue squamous carcinoma cell line Tca8113. AB - This study was designed to determine the effects of the recombinant adeno associated virus vector containing sense CD151 gene (rAAV-CD151) and antisense CD151 gene (rAAV-antiCD151) on the migration of Tca8113 cell. Functional fragment of CD151 gene was amplified by RT-PCR, and inserted into the vector pAAV in the sense direction and antisense direction, respectively. The rAAV-CD151 and rAAV antiCD151 were produced and the titers were determined by dot blot. The CD151, at protein level, was detected by Western blot. The Transwell chamber was used to detect the effects of the rAAV-CD151 and rAAV-antiCD151 on the tumor cell migration. The titers of the rAAV-CD151 and rAAV-antiCD151 were 2 x 10(11) pfu/ml and 1.0 x 10(11) pfu/ml, respectively. The expression of CD151 was increased by 108% in the cells transfected with rAAV-CD151 and decreased by 79% in the cells transfected with rAAV-antiCD151, as compared with non-transfected cells, respectively. The number of the migrating cells was significantly increased in the cells transfected with rAAV-CD151 (93.56 +/- 11.59) and decreased in the cells transfected with rAAV-antiCD151 (24.00 +/- 4.36) as compared with non transfected and rAAV-GFP transfected cells (53.00 +/- 6.56 and 46.00 +/- 7.00, P<0.05). It is an important molecular mechanism of the tumor metastasis that the overexpression of CD151 promotes the migration of the tumor cells. The rAAV antiCD151 is a novel tool, which can reduce the expression of CD151 and inhibit the migration of the tumor cells, and brings us a new approach of anti-sene gene therapy targeted at CD151 in human carcinoma. PMID- 15791840 TI - Study on the relationship between heat shock protein 70 and toll-like receptor-4 of monocytes. AB - To explore the relation between human heat shock protein 70 (hsp70) and TLR4 in human monocytes in vitro, human monocytes were stimulated with various concentrations of HSP70, and TNF-alpha production in supernatants was measured by ELISA. Pre-incubated with or without anti-TLR4 mAb, and stimulated with hsp70 (5.0 microg/ml), NF-kappaB p65 of human monocytes in different time points were detected by immunohistochemistry and monocyte surface expression of TLR4 was measured by flow cytometry. After the human monocytes were pre-incubated with various concentrations of anti-TLR4 and stimulated with hsp70 (5.0 microg/ml), TNF-alpha production in supernatants was measured. The results showed that hsp70 enhanced NF-kappaB activation, which was clearly inhibited by anti-TLR4, with the positive cell ratios being 67.44%, 39.17%, 31.56% and 28.05 %, respectively. TLR4 was rapidly down-regulated in the presence of hsp70. MFI of TLR4 on monocytes in different time points were 87.77 +/- 5.38, 78.16 +/- 6.01 and 45.17 +/- 4.97 (P<0.05), 26.98 +/- 5.83 (P<0.01), respectively. Moreover, hsp70-induced TNF alpha production by human monocytes was inhibited by anti-TLR4. It is suggested that TLR4 is involved in the hsp70-mediated activation of innate immunity. PMID- 15791841 TI - Effects of leptin on expression of acyl-coenzymea: cholesterol acyltransferases-1 in cultured human monocyte-macrophages. AB - To investigate the effects of leptin on expression of acyl-coenzymeA: cholesterol acyl-transferases-1 (ACAT-1) in monocyte-macrophage differentiation, human monocytic cells (THP-1) were cultured in RPMI 1640 and made to differentiate into macrophages under the incubation with phorbol myristate acetate (PMA) for 48 h. The cells were divided into 4 groups according to different intervention factors as follows: MCs cultured in RPM11640 medium with 10% FBS for 48 h served as MC group (control group), MCs cultured in medium with serum-free RPM11640 containing 5% BSA, 100 nmol/L PMA for 48 h as MP group, MCs cultured in RPMI1640 medium with 10% FBS, 10 micromol/ml leptin for 48 h as leptin-MC group, and MCs cultured in medium with serum-free RPMI1640 containing 5% BSA. 100 nmol/L PMA, and 10 micromol/ml leptin for 48 h as leptin-MP group. Immunocytochemistry, reverse transcription polymerase chain reaction (RT-PCR) and Western blot were performed, respectively, to observe the effects of leptin on expression of ACAT-1 in the monocyte-macrophage differentiation. Our results showed that expression of ACAT-1 protein and mRNA in MP-group is two times that in MC-group (P<0.05), and the expression of ACAT-1 protein and mRNA increased by up to 4 folds in leptin-MP group-as compared with that of MC group (P<0.01). Thus, our results support the idea that expression of ACAT-1 increases more in cultured human macrophages than in monocytes, and leptin can significantly promote ACAT-1 expression. It was concluded that high expression of ACAT-1 may accelerate the development of human atherogenesis, and leptin might participate in atherogenesis by increasing expression of ACAT-1. PMID- 15791842 TI - Angiogenic potency of bone marrow stromal cells improved by ex vivo hypoxia prestimulation. AB - To study the angiogenic potency of hypoxia-prestimulated bone marrow stromal cells (BMSCs) when transplanted into acute myocardial infarction models of rats. BMSCs were cultured under hypoxia condition for 24 h. Their expression of VEGF was investigated. The rat acute myocardial infarction models were made by coronary artery ligation and divided into 3 groups at random. In normoxia group, twice-passaged BMSCs were labeled with Bromodeoxyuridine (BrdU) and then implanted into the infarction regions and ischemic border of the recipients in 4 weeks. The rats in hypoxia group were implanted with hypoxia-prestimulated BMSCs. In control group, the model rats received only DMEM medium injection. Six-weeks after AMI, the infarction regions were examined to identify the angiogenesis and the expression of the VEGF. Our results showed that viable cells labeled with BrdU could be identified in the host hearts. The infarction regions in normoxia and hypoxia groups had a greater capillary density and increased VEGF expression than the regions in control group. The capillary density and VEGF expression in hypoxia group were higher than in normoxia group. It is concluded that the enhanced expression of VEGF in BMSCs could be induced by ex vivo hypoxia stimulation. BMSCs implantation promoted the angiogenesis in myocardial infarction tissue via supplying exogenic VEGF. Angiogenic potency of bone marrow stromal cells was improved by ex vivo hypoxia prestimulation though the enhanced VEGF expression. PMID- 15791843 TI - Effect of N-tosyl-L-phenylalanylchloromethyl ketone on tumor necrosis factor alpha -induced NF-kappaB activation and apoptosis in U937 cell line. AB - To investigate the effect of N-tosyl-L-phenylalanylchloromethyl ketone (TPCK) on tumor necrosis factor-alpha-induced NF-kappaB activation and apoptosis in U937 cell line, changes and subcellular localization of NF-kappaB/p65 and IkappaB alpha were observed by fluorescencemicroscopy and expression and degradation of IkappaB-alpha by flow cytometry. The apoptosis of U937 cells was measured by flow cytometry and electrophoresis of DNA. Immunolfluorescence assay showed that NF kappaB/p65, IkappaB-alpha only localized in cytoplasm. After TNF-alpha stimulation, p65 was localized only in nuclei, and IkappaB-alpha was only localized in cytoplasm and decreased. The changes of TNF-alpha stimulation were specifically inhibited by TPCK. Flow cytometry also revealed the downregulation of IkappaB-alpha protein during TNF-alpha-induced apoptosis and the down regulation was specifically inhibited by TPCK. Flow cytometry also showed the apoptosis of U937 cells after TNF-alpha induction. DNA ladder can be detected in cells treated by TNF-alpha. It is concluded that degradation of IkappaB-alpha protein and NF-kappaB/p65 translocation occur during TNF-alpha-induced apoptosis of U937 cells, suggesting the activation of NF-kappaB TPCK-sensitive protease plays an important role in the degradation of IkappaB-alpha protein induced by TNF-alpha in U937 cells. TPCK sensitive protease also plays an important role in the apoptosis of U937 cells induced by TNF-alpha. PMID- 15791845 TI - Effects of glucocorticoid on IL-13-induced Muc5ac expression in airways of mice. AB - To study the effects of glucocorticoid on the IL-13-induced Muc5ac expression in airways of mice, and investigate its role in mucus secretion of airways, 24 pathogen-free BALB/c mice were randomly divided into 3 groups. IL-13 group received an nasal instillation of 100 microg of recombinant murine IL-13 solution on days 1, 3 and 5. In dexamethasone group, dexamethasone (0.5 mg/kg) was administered intraperitoneally 24 h before and 1 h before the first instillation of IL-13 and on 4 consecutive days (day 0 to day 5, 6 consecutive days in total), while control group was not treated with IL-13 or dexamethasone. Bronchoalveolar lavage fluid (BALF) was collected and eosinophils were counted, and expression of Muc5ac mRNA and protein in lungs were detected by reverse transcription polymerase chain reaction (RT-PCR) technology and immunohistochemical assay respectively. Our results showed that the number of mice, with positve Muc5ac protein expression, expression of Muc5ac mRNA and eosinophils in BALF after IL-13 treatment were all significantly higher than that of control group (all P<0.01). Despite eosinophils reduced (P<0.01), the number of mice with positive Muc5ac protein expression, expression of Muc5ac mRNA afterdexamethasone treatment didn't decreas significantly as compared with that of IL-13 group. It is concluded that IL-13 can up-regulate the expression of Muc5ac mRNA and protein, which may play a pivotal role in the mucus overproduction of airways. Dexamethasone can suppress IL-13-induced eosinophilic infiltration in lung but can't inhibit the mucus overproduction. PMID- 15791844 TI - Regulation of histone acetylation and apoptosis by trichostatin in HL-60 cells. AB - In order to examine the strong anticancer action and low toxicity of Trichostatin A (TSA), the effect of TSA was examined on the growth inhibition, acetylation of histone H3 and apoptosis in HL-60 cells by employing MTT, immunocytochemical techniques, and Annexin-V-FITC/ PI assay. Our results showed that TSA could inhibit proliferation of HL- 60 cells in a time- and dose-dependent manner, and the IC50 at the 36th h was 100 ng/ml. The apoptosis-inducing effect of TSA on HL 60 cells was also time- and dose-dependent. But it didn't demonstrate apparent apoptosis induction in NPBMNCs within specific dose and time range. Both of the acetylation of histone H3 in HL-60 cells and NPBMNCs increased significantly (P<0.05) after treated with 100 ng/ml TSA for 4 h. However, there was no significant differences between the two groups (P>0.05). It is concluded that TSA can inhibit growth and induce apoptosis of HL-60 cells in a time- and dose dependent manner, and is able to selectively induce apoptosis in HL-60 cells but does not respond in NPBMNCs under the same conditions. The difference of TSA between HL-60 cells and NPBMNCs can't be explained by the regulation of histone acetylation. PMID- 15791846 TI - An integrated technique for identification of differential genes expressed in patients with cancer. AB - To develop a method for identification of differential gene expression between different cell populations, several convenient techniques of molecular biology, including subtractive hybridization, suppression PCR, T/A cloning and sequencing, were used to identify genes expressed differentially in CD45+ and CD45- cells isolated from U266 cell line of multiple myeloma. Our results showed that the levels of abundant genes scale down 20 times through subtractive hybridization. Plasmid DNA from CD45- cell clones was hybridized with forward or backward cDNA probes synthesized from CD45- and CD45- cells, respectively. A few of differentially expressed genes reconfirmed by RT-PCR were identified from 500 expressed clones of CD45+ cells. It is concluded that a strategy for gene expression identification developed from conventional molecular biological methods can be used in different laboratories. PMID- 15791848 TI - KLF6mRNA expression in primary hepatocellular carcinoma. AB - To investigate the expression of KLF6mRNA in primary hepatocellular carcinoma (HCC), nomal liver tissues and the tissues adjacent to the cancers, reverse transcription polymerase chain reaction (RT-PCR) was employed to investigate the expression of the KLF6 gene in HCC, the corresponding adjacent non-cancerous tissues and normal liver tissue. Our results showed that an amplified fragment of 427 bp DNA was detected in 18 of 19 (94.7%) adjacent non-cancerous tissues and normal liver tissue, and in 12 (85.7%) of 14 HCC. There were no significant differences in the levels of KLF6 mRNA between normal liver and liver tumors (P>0.05). It is concluded that KLF6 mRNA is generally expressed in HCC. PMID- 15791847 TI - Methodological studies on plasma endotoxin level and endotoxin inactivation capacity. AB - To establish stable methods for detecting plasma endotoxin level and endotoxin inactivation capacity in a normal population and general surgical patients and evaluate their perioperative changes. 50 healthy people and 50 patients receiving gastrointestinal operation were enrolled, their plasma endotoxin levels and plasma endotoxin inactivation capacity were assayed. Our results showed that plasma endotoxin levels were 0.044 +/- 0.009 EU/ml in the normal population and 0.044 +/- 0.023 EU/ml in the preoperative patients. Endotoxin level peaked 3 h after the operation (0.223 +/- 0.041 EU/ml), and then decreased rapidly on the first day after the operation (0.134 +/- 0.164 EU/ml). Endotoxin inactivation capacity also had the same time course as endotoxin level. Systemic inflammatory response syndrome and infection induced another elevation in the time course. It is concluded that establishing the endotoxin standard curve by using pyrogenic free water is better than by using plasma. Plasma endotoxin inactivation capacity can be used as an indirect indicator of postoperative immune depression. Plasma endotoxin level and endotoxin inactivation capacity peaked shortly after operation, indicating surgical stress is closely related with the changes. PMID- 15791849 TI - Expression and clinical significance of p27kip1 protein in primary liver cancer. AB - To investigate the expression and clinical significance of p27kip1 protein in primary liver cancer, the expression of p27kip1 protein and the relationship with clinicopathological factors were studied in primary liver cancer by using SABC immunohistochemical staining in specimens of 40 cases of primary liver cancer and 20 cases of liver cirrihosis. Our results showed that positive expression rate of p27kip1 protein in primary liver cancer was 37.5% (15/40), which was lower than that in benign lesion of liver 80.0% (16/20, P<0.01). The expression level of p27kip1 protein in primary liver cancer showed significant differences in tumor size, Edmonson histological grade, portal invasion, lymph node metastasis, TNM stage (P<0.05, for all), but not significantly correlated with patient's age and histological types. Log rank test showed that the p27kip1 expression was significantly related with prognosis of the patients (P<0.05), and the prognosis of the patients with p27kip1 positive expression was markedly better than that of those with p27kip1 negative expression. It is concluded that the expression of p27kip1 was significantly related clinicopathological factors of primary liver cancer. p27kip1 protein may be used as a novel tumor marker for primary liver cancer. PMID- 15791850 TI - Analysis of etiology and drug resistance of biliary infections. AB - The bile was collected from fro patients with biliary infections, with the bacterium isolated to study the sensitivity of each kind of the bacterium to several antibiotics in common use. Except G- bacterium, we also found some kinds of G+ bacterium in infection bile. G- bacterium were not sensitive to Clindamycin, G+ bacterium were sensitive to Ciprofloxacin. Escherichia coli, Xanthomonas maltophilia, Enterobacter cloacae, Pseudomonas aeruginosa were sensitive to Ampicillin. G+ bacterium were not sensitive to Azactam. Enterococcus faecalis, Enterococcus faecium, Enterobacter cloacae were not sensitive to Ceftazidime. Enterococcus faecalis, Staphylococcus coagulase negative, Staphylococcus epidermidis, Pseudomonas aeruginosa were not sensitive to Ceftriaxone Sodium. We didn't found any bacterium resistance Imipenem. The possibility of the existence of G+ bacterium as well as drug resistance should be considered n patients with biliary infections. The value of susceptibility test should be respected to avoid drug abuse of antibiotics. PMID- 15791851 TI - Expression and implication of hypoxia inducible factor-1alpha in prostate neoplasm. AB - To study the expression of hypoxia inducible factor-1alpha (HIF-1alpha) protein in prostate cancer (Pca) and its biological significance, the expression of HIF 1alpha was assayed by means of immunohistochemical technique in 42 prostate cancer, 12 prostatic intraepithelial neoplasm (PIN) and 9 normal prostate tissue (NP) specimens. Western blot was used to examine the expression of HIF-1alpha in prostate cancer cell line (PC-3M) induced by different oxygen tension. HIF-1alpha expression was positive in 33 Pca and 9 PIN specimens, and the positive rate of HIF-1alpha was higher in distant metastasis patients than in patients without metastasis of prostate cancer (P<0.05), while there was no expression of HIF 1alpha in NP. The level of HIF-1alpha in PC-3M significantly increased with the decrease of oxygen tension (P<0.01). Overexpression of HIF-1alpha is the preliminary event of the formation of Pca, which may induce carcinoma into malignant phenotype. Thus it may serve as an early diagnosis marker and the novel target for Pca treatment. PMID- 15791852 TI - Expression of vascular endothelial growth factor C and its correlation with lymph node metastasis in colorectal carcinoma. AB - To study the expression of vascular endothelial growth factor C (VEGF-C) in colorectal carcinoma and its relationship with lymph node metastasis, the expression of VEGF-C protein in colorectal carcinoma tissues obtained from 94 patients who underwent radical resection was immunohistochemically detected. Meanwhile, the expression of VEGF-C mRNA in 4 colorectal carcinoma cell lines was examined by reverse transcription polymerase chain reaction (RT-PCR). VEGF-C protein was found to be expressed in 53.2% of patients. The expression was more frequently detected in tumors with lymph node metastasis than in those without metastasis (P<0.01), and there was significant correlation between its expression and lymphatic invasion, TNM stage (P<0.01). However, no significant correlation was found between its expression and the age, gender, tumor location, depth of invasion and vascular invasion. 2 of the 4 colorectal carcinoma cell lines, including LoVo and LoVo-5FU, expressed VEGF-C mRNA. The expression of VEGF-C is closely related to lymph node metastasis, and it might take part in the tumor lymphangiogenesis. PMID- 15791853 TI - Ad/CMV- hTGF-beta1 treats rabbit intervertebral discs degeneration in vivo. AB - To investigate therapeutic efficiency of Ad/CMV- hTGF-beta1 gene for rabbit intervertebral disc degeneration model. 60 Japanese white rabbits were selected to form the 1.5-L6 Anterior-Lateral-Anulus-Fibrosus-Incision-Induced model in order to simulate human intervertebral disc degeneration. 36 rabbits, whose corresponding intervertebral discs were injected with 20 microl (10 x 10(6) pfu) of Ad/CMV- hTGF-beta1 gene, constituted the therapy group, 12 were injected with 20 microl (10 x 10(6) pfu)of Ad/CMV-LacZ gene as comparison group, while 12 were only injected with equivalent capacity of saline for empty comparison group, 3 weeks after injection, examples were taken for investigation of HE staining, MRI, Western Blotting and immunohistochemical research TGF-beta1. Wide distribution of TGF-beta1 was detected by immunohistochemical research in the degenerated annulus fibrosus after injection. Western Blotting research showed significant increase of TGF-beta1 content in intervertebral discs treated with TGF-beta1 gene than comparison groups. MRI signal transformed from low to comparatively high and that intervertebral disc pathological degree improved. Ad/CMV- hTGF-beta1 gene transfection is a potential method to increase TGF-beta1 content and reverse intervertebral disc degeneration. PMID- 15791854 TI - Effect of thiopental sodium on the release of glutamate and gamma-aminobutyric acid from rats prefrontal cortical synaptosomes. AB - To investigate the effect of thiopental sodium on the release of glutamate and gamma-aminobutyric acid (GABA) from synaptosomes in the prefrontal cortex, synaptosomes were made, the spontaneous release and the evoked release by 30 mmol/L KCl or 20 micromol/L veratridine of glutamate and GABA were performed under various concentrations of thiopental sodium (10-300 micromol/L), glutamate and GABA concentrations were determined by reversed-phase high-performance liquid chromatography. Our results showed that spontaneous release and evoked release of glutamate were significantly inhibited by 30 micromol/L, 100 micromol/L and 300 micromol/L thiopental sodium, IC50 of thiopental sodium was 25.8 +/- 2.3 micromol/L for the spontaneous release, 23.4 +/- 2.4 micromol/L for KCl-evoked release, and 24.3 +/- 1.8 micromol/L for veratridine-evoked release. But GABA spontaneous release and evoked release were unaffected. The study showed that thiopental sodium with clinically related concentrations could inhibit the release of glutamate, but had no effect on the release of GABA from rats prefrontal cortical synaptosomes. PMID- 15791855 TI - Effects of isoflurane on the actions of neuromuscular blockers on the muscle nicotine acetylcholine receptors. AB - In this study, we tested the hypothesis that volatile anesthetic enhancement of muscle relaxation is the result of combined drug effects on the nicotinic acetylcholine receptors. The poly A m RNA from muscle by isolation were microinjected into Xenopus oocytes for receptor expression. Concentration-effect curves for the inhibition of Ach-induced currents were established for vecuronium, rocuranium, and isoflurane. Subsequently, inhibitory effects of NDMRs were studied in the presence of the isoflurane at a concentration equivalent to half the concentration producing a 50% inhibition alone. All tested drugs produced rapid and readily reversible concentration-dependent inhibition. The 50% inhibitory concentration values were 889 micromol/L (95% CI: 711-1214 micromol). 33.4 micromol (95% CI: 27.1-41.7 nmol) and 9.2 nmol (95% CI: 7.9-12.3 nmol) for isoflurane. rocuranium and vecuronium, respectively. Coapplication of isoflurane significantly enhanced the inhibitory effects of rocuranium and vecuronium, and it was especially so at low concentration of NMDRs. Isoflurane increases the potency of NDMRs, possibly by enhancing antagonist affinity at the receptor site. PMID- 15791856 TI - Quantitative assessment of right ventricular systolic function by the analysis of right ventricular contrast time-intensity curve. AB - To study reliability and reliable indices of quantitative assessment of right ventricular systolic function by time-intensity curve (TIC) with right ventricular contrast, 5% sonicated human albumin was injected intravenously at a does of 0.08 ml/kg into 10 dogs at baseline status and cardiac insufficiency. Apical four-chamber view was observed for washin and washout of contrast agent from right ventricle. The parameters of TIC were obtained by curve fitting. The differences of parameters were analyzed in different states of cardiac functions. Among the parameters derived from TIC, the time constant (k) was decreased significantly with decline of cardiac function (P<0.001). But half-time of decent of peak intensity (HT) and mean-transit-time (MTT) of washout were increased significantly (P<0.001). The k was strongly related to cardiac output of right ventricle (CO) and ejection fraction (EF) of left ventricle and fractional shortening (FS) of left ventricle. Right ventricular systolic function could be assessed reliably by the parameters derived from TIC with right ventricular contrast echocardiography. The k, HT and MTT are reliable indices for quantitative assessment of right ventricular systolic function. PMID- 15791857 TI - [Comparison of gradient echo and echo planar imaging measurements for the calculation of regional blood volume maps]. AB - The purpose of this study was to evaluate relative regional blood volume (rCBV) in meningioma and compare the utility of reconstruction using both gradient-echo sequence and echo-planar-imaging (EPI) sequence. Eighteen patients with meningiomas were studied on a Siemens 1.5-T scanner. During the gradient-echo sequence (n=12) and EPI sequence (n=6), a bolus (0.2 mmol/kg) of Gd-DTPA was injected mechanically with a flow rate of 5 ml/second. Image processing of dynamic data was performed on a pixel-by-pixel basis. The ratio of tumor rCBV/gray matter rCBV was 3.01 +/- 1.18 (3.07 +/- 1.39 in gradient sequence and 2.84 +/- 0.94 in EPI sequence). The ratio of gray matter/white matter as the reference tissue had a mean of 2.79 +/- 0.76 using the FLASH sequence, and a mean of 3.04 +/- 1.31 using EPI. These differences were not statistically signifcant (P>0.5, t-test). According to the ratio of tumor rCBV/gray matter rCBV, a mean value, 14.5 ml/100 g, of rCBV in meningiomas was calculated. Compared with gray matter, increased inhomogeneous rCBV was observed in meningioma. Based on the two different sequences, no bias can be observed in our rCBV reconstruction. PMID- 15791858 TI - Dynamic changes of the CT perfusion parameters in the embolic model of cerebral ischemia. AB - To study the dynamic changes of CT perfusion parameters during the first 12 h in the embolic cerebral ischemia models. Local cerebral ischemia model were established in 7 New Zealand white rabbits. All CT scans were performed with a GE Lightspeed 16 multislice CT. Following the baseline scan, further CT perfusion scans were performed at the same locations 20 min, 1-6 h and 8, 10 and 12 h after the embolus delivery. Maps of all parameters were obtained by CT perfusion software at each time point. The brains, taken 12 h after the scan, were sliced corresponding to the positions of the CT slices and stained by 2,3,5 triphenyltetrazolium chloride (TTC). On the basis of the TTC results, the ischemic sides were divided into 3 regions: core, penumbra and the relatively normal region. The changes of all parameters were then divided into 3 stages. In the first two hours (the first stage), the CBV dropped more remarkably in the core than in the penumbra but rose slightly in the relatively normal region while the CBF decreased and MTT, TTP extended in all regions to varying degrees. In the 2nd-5th h (the second stage), all the parameters fluctuated slightly around a certain level. In the 5th-12th h (the third stage), the CBV and CBF dropped, and MTT and TTP were prolonged or shortened slightly in the core and penumbra though much notably in the former while the CBV, CBF rose and MTT, TTP were shortened remarkably in the relatively normal region. We experimentally demonstrated that the location and extent of cerebral ischemia could be accurately assessed by CT perfusion imaging. The pathophysiology of the ischemia could be reflected by the CT perfusion to varying degrees. PMID- 15791859 TI - Use of PEI-coated magnetic iron oxide nanoparticles as gene vectors. AB - To evaluate the feasibility of using polyethyleneimine (PEI) coated magnetic iron.oxide nanoparticles (polyMAG-1000) as gene vectors. The surface characteristics of the nanoparticles were observed with scanning electron microscopy. The ability of the nanoparticles to combine with and protect DNA was investigated at different PH values after polyMAG-1000 and DNA were combined in different ratios. The nanoparticles were tested as gene vectors with in vitro transfection models. Under the scanning electron microscope the nanoparticles were about 100 nm in diameter. The nanoparticles could bind and condense DNA under acid, neutral and alkaline conditions, and they could transfer genes into cells and express green fluorescent proteins (GFP). The transfection efficiency was highest (51%) when the ratio of nanoparticles to DNA was 1:1 (v:w). In that ratio, the difference in transfection efficiency was marked depending on whether a magnetic field was present or not: about 10% when it was absent but 51% when it was present. The magnetic iron oxide nanoparticles coated with PEI may potentially be used as gene vectors. PMID- 15791860 TI - Expression of resistin mRNA in adipose tissue of rat model with polycystic ovarian syndrome and its implication. AB - To investigate the relationship between the insulin resistance (IR) of polycystic ovarian syndrome (PCOS) rat model induced by dehydroeplandrosterone (DHEA) and hormonal changes in the ovarium and the resistin mRNA levels in adipose tissue, 21-day-old female SD rats were divided into two groups in pairs. The rats in group 1 were injected daily (s.c.) with DHEA for up to 20 days and the rats in group 2 injected with oil at the same time. Ovarian weight, serum insulin levels and sex hormone levels in rat blood of both groups were determined. Oral glucose tolerance tests, light microscopic and electronic microscopic examination were performed. The levels of resistin mRNA in adipose tissue were measured by reverse transcription-polymerase chain reaction (RT-PCR). Our results showed that the ovarian weight in group 1 was greater than that in group 2 (P<0.05). The ovaria in group 1 showed multiple follicular cysts, The serum testeosterone and etrasdiol concentration were significantly higher in group 1 than those in group 2 (P<0.001 and P<0.05 respectively), so as the fasting serum glucose (P<0.001) and fasting serum insulin (P<0.05). The value of 1/FINS x FGC was significantly higher in group 1 than that in group 2 (P<0.001). Moreover, the resistin mRNA level of white adipose tissue in the DHEA-induced group was significantly higher than that in the control rats (P<0.05). It is concluded that the DHEA-induced PCOS rat models were similar to those of the patients with PCOS, and the IR was observed. Resistin secreted by adipose tissue may mediate IR in PCOS, and it is likely involved in the pathogenesis and development of PCOS. PMID- 15791861 TI - Transcription and translation of Dickkopf-1 in endometrium of pregnant mice during the peri-implantation period. AB - To study the expression of Dickkopf-1 (DKK-1) in endometrium of pregnant mice during the peri-implantation period and the role of DKK-1 during the embryo implantation in mice. Immunohistochemical technique was employed to determine the location of DKK-1 protein in endometrium, and semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) was utilized to determine the levels of DKK-1 mRNA. Our results showed that the expressions of DKK1 mRNA and protein were higher in experimental groups than in control group (P<0.01) and it increased significantly on day 3 and reached its peak on day 4, and then decreased gradually on day 5-7. The levels of DKK-1 mRNA and protein on day 4 was significantly higher than those of other groups (P<0.01). It is concluded that DKK-1 probably plays an important role in signal transudation of embryo implantation and its high expression indicates the opening of implantation window. PMID- 15791863 TI - Immunohistochemical analysis of TNF-alpha and HSP-60 in women with tubal factor infertility associated with Chlamydia trachomatis. AB - To explore the roles of tumor necrosis factor-alpha (TNF-alpha) and heat shock protein 60 (HSP-60) in women with tubal factor infertility (TFI) associated with Chlamydia trachomatis, and to determine the mechanisms of fallopian adhesions in Chlamydia trachomatis (CT) infections, the expressions of TNF-alpha and HSP-60 were quantitatively determined in 60 cases of TFI and 30 controls by immunohistochemical technique. The patients with TFI were further divided into group A and group B according to the CT-DNA of cervical specimens of PCR. The quantitative analysis was conducted by employing computerized image analysis system. It is found that the expressions of TNF-alpha and HSP-60 were much higher in TFI patients than those of controls. Among CT-HSP responders, a stronger expression was correlated with more severe salpingeal pathology. It is concluded that TNF-alpha and HSP-60 play very important roles in fallopian tube adhesion and occlusion in TFI due to CT infection. PMID- 15791862 TI - The correlation between prognosis of HCMV infection and zinc in mature women. AB - The aim of the study was to investigate the correlation between prognosis of human cytomegalovirus (HCMV) infection and zinc in mature women, in order to explore the effect of serum zinc on HCMV infection. 900 mature woman were examined for zinc level and serum HCMV specific antibodies (IgG and IgM). 63 HCMV IgM positive cases were divided into 3 subgroups according to their serum zinc level, and HCMV-IgM was observed for conversion after treatment with a Chinese traditional medicine (Jinyebadu). Our results showed that the mean concentration of zinc in serum was significantly lower in HCMV-IgM(+) group than that in HCMV IgM(-) group (P<0.001). A positive correlation was found between the response of mature females with HCMV infection to the treatment and the levels of serum zinc (P<0.001). The response of mature women with HCMV is poor when the serum zinc is lower than 0.70. PMID- 15791864 TI - Expression of tissue transglutaminase in cultured bovine trabecluar meshwork cells. AB - To study whether cultured bovine trabecluar meshwork cells (BTMC) are capable of expressing tTG in protein and at mRNA level, BTMC were cultured in vitro and passaged three times, then the cells were transferred onto or cultured on sterile cover or submitted to isolation of RNA with Trizol, and the expression of tTG was detected by immunohistochemical technique and reverse transcription polymerase chain reaction (RT-PCR) respectively. Our results showed that tTG immunostaining was positive in the cytoplasm and rarely in the nucleus of cultured BTMC. No immunostaining was seen in the negative control. Moreover, a single RT-PCR amplified product whose sequence and size were in accordance with our known parameters was obtained. The expression of tTG in cultured BTMC was confirmed in protein and at mRNA level. BMTC is available more readily for the investigation of the relationship between tTG and primary open-angle glaucoma. PMID- 15791865 TI - Expression of hypoxia inducible factor-1alpha and its relationship to apoptosis and proliferation in human laryngeal squamous cell carcinoma. AB - To investigate the expression of hypoxia inducible factor-1 alpha (HIF-1alpha) and its relationship to apoptosis and proliferation in laryngeal squamous cell carcinoma (LSCC), immunohistochemical method was used to detect the expression of HIF-1alpha and PCNA. Tunnel technique was used to detect in situ cell apoptosis in LSCC. Our results showed that the expression of HIF-1alpha was related to the clinical stages of cancer and lymph node metastasis (P<0.05). The relationship between HIF-1alpha and PCNA was statistically significant (P<0.05) and no relationship was found between HIF-1alpha and apoptosis (P>0.05) It is concluded that HIF-1alpha plays a role in the carcinogenesis of laryngeal carcinoma and is correlated with proliferation, but bears no relationship with the apoptosis of tumor cells in LSCC. PMID- 15791866 TI - The expression of toll-like receptor 2 and 4 mRNA in local tissues of model of oropharyngeal candidiasis in mice. AB - To investigate the expression of Toll-like receptor (TLR) 2 and 4 mRNA in local tissues of model of oropharyngeal candidiasis in mice and to explore the potential role of TLR2 and TLR4 in earlier period of immune response, a murine model of oropharyngeal candidiasis inoculated by cotton wool balls saturated with Candida albicans was established. Mice were sacrificed at the indicated time points and the oropharyngeal tissues were excised. The expression of TLR2 and TLR4 mRNA was detected by RT-PCR. The results showed that low level of TLR2/4 mRNA could be detected in oropharyngeal tissues, but they were markedly up regulated 6 h after inoculation, peaking after 12-24 h. Tissue TLR4 mRNA was gradually down-regulated 24-48 h, while TLR2 mRNA levels remained high up to the 72nd h. These data suggested that oropharyngeal infection of Candida albicans could result in up-regulation of TLR2/4 mRNA expression in local tissues, which might play important roles in earlier period of immune response. PMID- 15791867 TI - Clinical identification of common species of dermatophytes by PCR and PCR-RFLP. AB - To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme Hinc II. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans. From the restriction profiles of Hinc II, 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCR-RFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase II gene is rapid and efficient. PMID- 15791868 TI - Fatigue and doctors. PMID- 15791869 TI - Modernising Medical Careers or 'modernising clinical careers'? PMID- 15791870 TI - Implications of the new curriculum for medical trainers. AB - The increasing complexity of the role of trainer, highlighted in the expectations of the Modernising Medical Careers curriculum, currently out for consultation, exposes inadequacies in the current system. Changes in the supervisory structure are required and further training is essential if consultants are to fulfil this role adequately. PMID- 15791871 TI - Modernising Medical Careers: effect on NHS service delivery. AB - Modernising Medical Careers will be introduced from August 2005 and will significantly change the system of education and training for doctors in the UK. The impact of these changes on NHS service delivery needs to be fully understood and is discussed in this article. PMID- 15791872 TI - The Postgraduate Medical Education and Training Board. AB - The Postgraduate Medical Education and Training Board (PMETB) is an independent regulatory body which sets standards, approves, quality assures and evaluates postgraduate medical education and training. The PMETB is currently preparing to take over as the single competent authority for postgraduate medical education and training in September 2005. PMID- 15791873 TI - Parenteral nutrition: multidisciplinary management. AB - Parenteral nutrition may be needed to give nutritional support to patients with severe acute (temporary or reversible) or chronic intestinal failure. Parenteral nutrition needs to be given only by health workers trained in its use otherwise life-threatening complications (especially sepsis) may occur. PMID- 15791874 TI - Paediatric sepsis and multiple organ failure. AB - Mortality from paediatric sepsis and multiple organ failure has fallen over the last half century, but both remain significant causes of death and morbidity among children and infants in the intensive care unit. This article provides a basic guide for front-line clinicians involved in paediatric resuscitation, addressing nomenclature, epidemiology and suggestions for early treatment. PMID- 15791875 TI - Does prolapse surgery address the symptoms that concern women? AB - Clinicians are constantly being challenged to improve services and the care provided for individual patients. This article looks at the current approach to the management of uterovaginal prolapse in women and explores new developments in the assessment of prolapse and related pelvic floor disorders. PMID- 15791876 TI - Understanding psoriatic arthritis. AB - Previously, psoriatic arthritis was considered an uncommon, generally benign variant of ankylosing spondylitis or rheumatoid arthritis, with few therapeutic options. It is now recognized as a distinct, potentially serious disorder that can be effectively managed with a number of new agents. PMID- 15791877 TI - PRHO views on the usefulness of a pilot ward simulation exercise. AB - There are concerns that pre-registration house officers (PRHOs) lack the expertise to initially manage acutely-ill patients. Simulation can support them in this role by providing a safe yet authentic setting. This paper shares PRHOs' views of a ward simulation exercise. The positive feedback has resulted in further development of the exercise to support PRHOs who have given cause for concern in their practice. PMID- 15791878 TI - Boerhaave's syndrome secondary to epigastric hernia. PMID- 15791879 TI - Cavitating lung disease in ankylosing spondylitis may have other causes. PMID- 15791880 TI - Importance of haemodynamic monitoring in cardiac tamponade. PMID- 15791881 TI - Bladder extrophy. PMID- 15791883 TI - Antibiotic prescribing as a risk factor for MRSA. AB - There is accumulating evidence to support the role of specific antibiotic classes as risk factors for selecting meticillin-resistant Staphylococcus aureus (MRSA). Crucially, however, controlled studies of antimicrobial prescribing interventions are lacking. The evidence for the association between antimicrobial prescribing and expansion of MRSA is reviewed here. PMID- 15791882 TI - Secondary syphilis: a multisystem disease not to be forgotten. PMID- 15791884 TI - Sub-specialization in general surgery--the end of the 'general' surgeon? PMID- 15791885 TI - Managing acute cardiac patients in and out of the hospital setting. PMID- 15791886 TI - Case report: Wegener's granulomatosis presents as pulmonary infection. PMID- 15791887 TI - Diagnosis and treatment of group A pharyngitis strep. PMID- 15791888 TI - Dietary supplement use in children: concerns of efficacy and safety. PMID- 15791889 TI - Mildly elevated liver transaminase levels in the asymptomatic patient. AB - Mild elevations in liver chemistry tests such as alanine transaminase and aspartate transaminase can reveal serious underlying conditions or have transient and benign etiologies. Potential causes of liver transaminase elevations include viral hepatitis, alcohol use, medication use, steatosis or steatohepatitis, and cirrhosis. The history should be thorough, with special attention given to the use of medications, vitamins, herbs, drugs, and alcohol; family history; and any history of blood-product transfusions. Other common health conditions, such as diabetes, heart disease, and thyroid disease, can cause or augment liver transaminase elevations. The recent American Gastroenterological Association guideline regarding the evaluation and management of abnormal liver chemistry tests proposes a practical, algorithmic approach when the history and physical examination do not reveal the cause. In addition to liver chemistries, an initial serologic evaluation includes a prothrombin time; albumin; complete blood count with platelets; hepatitis A, B, and C serologies; and iron studies. Depending on the etiology, management strategies may include cessation of alcohol use, attention to medications, control of diabetes, and modification of lifestyle factors such as obesity. If elevations persist after an appropriate period of observation, further testing may include ultrasonography and other serum studies. In some cases, biopsy may be indicated. PMID- 15791890 TI - Treatment of vertigo. AB - Vertigo is the illusion of motion, usually rotational motion. As patients age, vertigo becomes an increasingly common presenting complaint. The most common causes of this condition are benign paroxysmal positional vertigo, acute vestibular neuronitis or labyrinthitis, Meniere's disease, migraine, and anxiety disorders. Less common causes include vertebrobasilar ischemia and retrocochlear tumors. The distinction between peripheral and central vertigo usually can be made clinically and guides management decisions. Most patients with vertigo do not require extensive diagnostic testing and can be treated in the primary care setting. Benign paroxysmal positional vertigo usually improves with a canalith repositioning procedure. Acute vestibular neuronitis or labyrinthitis improves with initial stabilizing measures and a vestibular suppressant medication, followed by vestibular rehabilitation exercises. Meniere's disease often responds to the combination of a low-salt diet and diuretics. Vertiginous migraine headaches generally improve with dietary changes, a tricyclic antidepressant, and a beta blocker or calcium channel blocker. Vertigo associated with anxiety usually responds to a selective serotonin reuptake inhibitor. PMID- 15791891 TI - Treatment of cholesterol abnormalities. AB - Cardiovascular disease and its subset coronary heart disease are leading causes of morbidity and mortality in the United States and worldwide. In general, higher levels of low-density lipoprotein cholesterol are associated with an increased risk of coronary heart disease, myocardial infarction, and stroke. Reducing dietary fat can improve total cholesterol levels, but consequent reductions in cardiovascular outcomes are not well documented. The Mediterranean diet is the only dietary intervention associated with a reduction in all-cause mortality. Treatment with cholesterol-lowering medications decreases the rate of cardiovascular events, but a reduction in all-cause mortality with these agents has been found only in patients with pre-existing coronary heart disease. Drug treatment in patients with a history of heart disease and average-to-high cholesterol levels can decrease the risk for stroke. In patients with peripheral vascular disease, treatment of elevated cholesterol levels may slow disease progression. PMID- 15791892 TI - Urinalysis: a comprehensive review. AB - A complete urinalysis includes physical, chemical, and microscopic examinations. Midstream clean collection is acceptable in most situations, but the specimen should be examined within two hours of collection. Cloudy urine often is a result of precipitated phosphate crystals in alkaline urine, but pyuria also can be the cause. A strong odor may be the result of a concentrated specimen rather than a urinary tract infection. Dipstick urinalysis is convenient, but false-positive and false-negative results can occur. Specific gravity provides a reliable assessment of the patient's hydration status. Microhematuria has a range of causes, from benign to life threatening. Glomerular, renal, and urologic causes of microhematuria often can be differentiated by other elements of the urinalysis. Although transient proteinuria typically is a benign condition, persistent proteinuria requires further work-up. Uncomplicated urinary tract infections diagnosed by positive leukocyte esterase and nitrite tests can be treated without culture. PMID- 15791893 TI - Evaluating the patient with a knee injury. PMID- 15791894 TI - An ulcerating foot lesion. PMID- 15791895 TI - Summary health statistics for the U.S. population: National Health Interview Survey, 2001. AB - OBJECTIVES: This report presents health statistics from the 2001 National Health Interview Survey for the civilian noninstitutionalized population of the United States, classified by age, sex, race and Hispanic or Latino origin, family income, poverty status, education, place of residence, region of residence, and, where appropriate, health insurance coverage. The topics covered are health status and limitations in activities, special education or early intervention services, injuries and poisonings, health care access and utilization, and health insurance coverage. SOURCE OF DATA: The NHIS is a household, multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control's National Center for Health Statistics. Household interviews were completed for 100,760 persons living in 38,932 households, reflecting a household response rate of 89%. SELECTED HIGHLIGHTS: Nearly 7 in 10 persons were in excellent or very good health in 2001. About 33 million persons (12%) were limited in their usual activities due to one or more chronic health conditions, and about 4 million persons (2%) required the help of another person with activities of daily living. Persons with the least education and the lowest incomes were the most likely to be limited in their ability to work. About 6% of children received special education or early intervention services. The three leading causes of medically attended injury and poisoning episodes were falls, transportation, and overexertion. Among persons under age 65 years, about 39 million (16%) did not have any health insurance coverage. The most common reason for lacking health insurance was cost, followed by a change in employment. PMID- 15791896 TI - Health behaviors of adults: United States, 1999-2001. AB - This report presents prevalence estimates for key indicators of alcohol use, cigarette smoking, leisure-time physical activity, and body weight status among U.S. adults, using data from the 1999-2001 National Health Interview Surveys (NHIS). The NHIS is conducted annually by the Centers for Disease Control and Prevention's National Center for Health Statistics. Estimates are shown for several sociodemographic subgroups for both sexes combined and for men and women separately. Subgroups are compared in terms of prevalence of healthy and unhealthy behaviors. Methods Data for the U.S. civilian noninstitutionalized population were collected using computer-assisted personal interviews (CAPI). Questions on health behaviors were asked in the Sample Adult component of the basic core questionnaire. All data were self-reported. This report is based on a total of 96,501 completed interviews with sample adults aged 18 years and over, representing an overall sample adult response rate of 71.8% for the 3 years combined. Statistics were age adjusted to the 2000 U.S. standard population. Results Overall, 6 in 10 U.S. adults were current drinkers in 1999-2001; about 1 in 4 adults (23.1%) were lifetime abstainers. About 1 in 4 adults (23.1%) were current smokers and over one-half of adults (54.3%) had never smoked cigarettes. About 6 in 10 adults engaged in at least some leisure-time physical activity with about 3 in 10 regularly engaging in such activities. About 6 in 10 adults were overweight or obese (BMI > or = 25), with 4 in 10 adults having a healthy weight. PMID- 15791897 TI - Summary health statistics for U.S. children: National Health Interview Survey, 2002. AB - OBJECTIVES: This report presents both age-adjusted and unadjusted statistics from the 2002 National Health Interview Survey on selected health measures for children under 18 years of age, classified by sex, age, race, Hispanic origin, family structure, parent's education, family income, poverty status, health insurance coverage, place of residence, region, and current health status. The topics covered are asthma, allergies, learning disability, Attention Deficit Hyperactivity Disorder (ADHD), use of prescription medication, respondent assessed health status, school-loss days, usual place of health care, time since last contact with a health care professional, unmet dental needs, time since last dental contact, and selected measures of health care access. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics and is representative of the civilian noninstitutionalized population of the United States. Data are collected during face-to-face interviews with adults present at the time of interview. Information about children is collected for one randomly selected child per family in face-to face interviews with an adult proxy respondent familiar with the child's health. SELECTED HIGHLIGHTS: In 2002, most U.S. children under 18 years of age had excellent or very good health (84%). However, 10% of children had no health insurance coverage, and 5% of children had no usual place of health care. Twelve percent of children had ever been diagnosed with asthma. An estimated 8% of children 3-17 years of age had a learning disability, and an estimated 7% of children had ADHD. PMID- 15791898 TI - [A review of epidemiological studies about pulse wave velocity for prevention of cardiovascular disease]. AB - Early arteriosclerosis can be prevented by medication and life style changes. However, late cardiac disease, called cardiovascular disease, could be fatal or have severe sequelae. Therefore, it is important to develop a method for early diagnosis. Since no symptoms become evident until endothelial/vascular dysfunction, a simple and sensitive diagnostic method using a biological parameter needs to be developed for prevention and medical treatment. Pulse wave velocity (PWV) is a noninvasive and simple measure for evaluating arterial wall stiffness and several studies have indicated this is a good marker for vascular damage, although not all results were consistent. This review concerns the following three aspects: 1. PWV as a marker of vascular damage 2. PWV as a prognostic predictor 3. Brachial-ankle PWV (baPWV) Blood pressure is strongly related with PWV, along with BMI, fasting blood glucose, and cholesterol. However, no relation was rated cigarette smoking. While many reports have shown that rise of PWV can serve as a prognostic predictor of disease, the situation with baPWV remains unclear. It is to be expected that improvements in methods for PWV will greatly contribute to primary and secondary prevention in the future. PMID- 15791899 TI - [Evaluation of service screening for breast cancer by clinical breast examination using regional cancer registry data]. AB - MATERIALS: (1) Screening test: The study utilized data from a regional cancer screening center from April 1, 1997, to December 31, 1998. A total of 51,700 women underwent screening at this center during this period. All incident cases of breast cancer in these women were derived from the files of the Yamagata Prefectural Cancer registry and sensitivity, specificity and positive predictive value were then estimated. False negative cases were defined as occurring in women who tested negative but were registered as having breast cancer in the cancer registry within 12 months after screening. (2) Effectiveness of screening: The target population comprised female cases with breast cancer registered in the Yamagata Prefectural Cancer Registry from January 1, 1989 to December 31, 1998. During this period, a total of 2,746 cases were registered. Survival probabilities for breast cancer cases were estimated according to method of detection using Kaplan-Meier method. Overall survival probability of the screen detected group was compared with the not screen-detected group using the log-rank test. Point estimates of 5- and 9-year survival rates between groups were compared using z statistics. RESULTS: (1) Screening detected 27 breast cancer cases, while 31 false negative cases were identified. Sensitivity, specificity and positive predictive value were 46.6% (95% confidence interval (95% CI), 33.3 60.1%), 97.3% (95% CI, 97.2-97.5%) and 1.9% (95% CI, 1.3-2.8%), respectively. (2) Overall survival rate for the screen-detected group was significantly higher than that for the not screen-detected group (P<0.001), with 11.6% (95% CI, 8.0-15.2%) and 13.1% (95% CI: 5.3-20.8%) differences in 5-year and 9-year survival probabilities, respectively. CONCLUSION: Cancer screening by clinical breast examination in Yamagata has improved survival from breast cancer for females. However, the screening sensitivity is insufficient for effective mass screening purposes. PMID- 15791900 TI - [An estimation of the prevalence of excessive daytime sleepiness based on age and sex distribution of epworth sleepiness scale scores: a population based survey]. AB - OBJECTIVES: To describe the distribution of subjective daytime sleepiness among local residents using the Epworth Sleepiness Scale (ESS) and estimate the prevalence of excessive daytime sleepiness (EDS) in a general Japanese adult population. METHODS: Subjects consisted of all residents aged 20 years and over from a self-governing body of approximately 10,000 people located in the Hokkaido region. Questionnaires, which included a Japanese version of the ESS, were distributed and later collected by a health promoter who visited subjects' residences between October and December 2000. Subjects of the analysis were restricted to those who answered five items or more out of a total of eight items. Means and standard deviations of the ESS global score were calculated for each age group by gender. And differences with reference to sex and age were examined by ANOVA. The prevalence of EDS in the Japanese general population was estimated from this study's results by direct age adjustment using the Japanese census figures for 2000. Factors related to EDS were also examined. RESULTS: A total of 5,327 residents responded to the survey (86.0%). Of these, 4,412 (71.2%) were entered for analysis. The average (+/- standard deviation) ESS global score were with 5.18 +/- 3.75 (males 5.25 +/- 3.89, females 5.12 +/- 3.75). Differing significantly age, regardless of sex (P<0001). From the overall prevalence of EDS of 9.2% (males 9.6%, females 8.8%), the estimated prevalence in the Japanese general population was to be 8.9% (males 9.3%, females 8.4%). EDS was found to be related to age, sleep duration under six hours and to snoring (P=0.002, P=0.008 and P<0.001). CONCLUSION: This study provided baseline data for the distribution by sex and age group of subjective daytime sleepiness in a community using the ESS and was used to generate the first ever estimated prevalence of EDS in the general Japanese adult population. The finding should prove useful for clinicians and researchers interested in (1) screening for daytime sleepiness, (2) evaluating changes between pre and post-interventions, (3) comparing epidemiological findings across studies, and lastly, (4) making of health policy. Daytime sleepiness estimated by ESS differs with the sex and age, possibly related to bio-medical and socio-medical factors. Further research is need to detail this problem. PMID- 15791901 TI - [Social rehabilitation experiences of people with a history of Hansen's disease: interviews of readmitted residents in a sanatorium]. AB - PURPOSE: This study focused on the reflections of people with a history of Hansen's disease who once experienced social life and then returned to their sanatorium again. The purpose was to clarify what they think about their experiences of social life outside of institutions. METHOD: Study participants were people with a history of Hansen's disease who had experienced social life outside of the sanatorium and are now living in a national sanatorium with good ADL. The study was based on a semi-structured interview, the data being categorized with the focus on "situations or topics affect by Hansen's disease". Thirteen people agreed to participate in the study. RESULTS AND DISCUSSION: Six categories were abstracted: "maintaining a good condition," "anxiety about relapse or illness," "medical service," "coping with Hansen's disease in social life," "relationships with others," and "means of livelihood." Hansen's disease affected their lives in 2 aspects; one was "sawagu (recurrence of the disease)" and the other "kakusu (concealing the disease)." "Sawagu" was related to "anxiety of the disease" and "intention of maintaining a good condition." "Kakusu" affected the way of "coping with the disease in social life" and "the relationship with others." "Medical service" was important for both "sawagu" and "kakusu." Patients were visiting hospitals and taking medicine to avoid "sawagu," while trying to "kakusu" when they consulted with physicians. Similarly, both "sawagu" and "kakusu" interacted with each other when the participants needed to work and earn a living. All participants looked back at their social life as a "good experience" because they were satisfied with the sense of accomplishment or fulfillment the experience had given them. CONCLUSION: Participants of the study looked back at their social life as a "good experience." Hansen's disease affected their social life in 2 aspects; "sawagu" and "kakusu." It is necessary to alleviate difficulties with both of these so that people with stigmatized disease such as Hansen's disease can maintain normal lives in society. PMID- 15791902 TI - [How has medical inspection by Public Health Center been accepted for medical care institutions?]. PMID- 15791903 TI - [A nationwide survey of trends in prospective payment system for psychiatric inpatient services]. PMID- 15791905 TI - [A comparative study of the physical conditions of elderly people with care needs receiving rehabilitation services from a nurse or a physiotherapist from a visiting nurse service station]. AB - PURPOSE: The purpose of this study was to examine the roles of rehabilitation by a nurse, comparing the attributes of utilization of care services and physical conditions of elderly people receiving rehabilitation services from a nurse or a physiotherapist. METHOD: Two hundred and fifty four care receivers at the Saiseikai visiting nurse service station, Shiga Prefecture, were interviewed by a nurse or a physiotherapist. They were divided into two groups: 1) receivers of rehabilitation services by a nurse (RRSN group), and 2) receivers of rehabilitation services by a physiotherapist (RRSP group). The subjects were matched for gender and age, and 36 participants for each of the two groups were included in the analysis. Level of dementia, activities of daily living (ADL; Barthel Index), instrumental activities of daily living (IADL), the Glasgow Coma Scale (GCS) and use of visiting services were assessed in the interview. Analysis of variance and the chi2 test were used to compare values for the two groups. RESULTS: Level of dementia in the RRSN group was significantly severe than in the RRSP group (P<0.05). The number of patients who are required to be treated with visiting medical services was significantly more in the RRSN group (P<0.05). Physical status in the RRSN group was significantly lower for ADL, GCS than in the RRSP group. IADL of males in the RRSN group was significantly lower. CONCLUSIONS: Thus, we conclude that it is important for nurses to make opportunities to visit elderly people with physiotherapists to assess their physical conditions. PMID- 15791904 TI - [Memory complaints among community-dwelling elderly in Japan: comprehensive health examination for the community elderly for prevention of the geriatric syndrome and a bed-ridden state ("Otasha-kenshin") part III]. AB - PURPOSE: Previous studies have indicated that memory complaints may predict cognitive decline and dementia among the elderly. The present study was therefore conducted to clarify memory complaint characteristics among elderly dwelling in an urban community in Japan. METHOD: The participants analyzed in the present study were 453 men and 385 women aged 70 to 84 years living in an urban Japanese community. Data on problems related to memory complaints, cognitive decline (below 24 points on Mini-Mental State Examination), depression (measured by Mini International Neuropsychiatric Interview), hearing and vision problems, I-ADL (measured by TMIG Index of Competence), self-rated health, age, sex, and years of education were collected at a comprehensive mass health examination for the elderly ("Otasha-kenshin"). RESULTS AND DISCUSSION: Twenty-seven percent of male respondents and 32% of female respondents reported having current trouble remembering things (reported as "frequently" or "sometimes"). We collected specific descriptions of the memory complaint difficulties the subjects were experiencing. A quarter of the responses indicated problems with forgetting persons' names, a fifth with forgetting where things had been left, 15% with leaving things behind, and a quarter with prospective memory failure. The results of multivariate logistic regression analysis to explore correlates showed that in men self-rated health and cognitive decline, and in women hearing problems and self-rated health were significantly and independently related to the memory complaint. These findings suggest that in addition to cognitive decline, self rated health and hearing problems may influence memory complaints, and that memory complaints in men may be a reliable, simple indicator of cognitive decline. We now need to carry out a longitudinal study to clarify predictive validity. PMID- 15791906 TI - [Risk factors for presence of complications of the mechanical ventilatory assistance in the newborn]. AB - BACKGROUND: The mechanical ventilatory assistance (MVA) is a procedure that is used very often in the neonatal intensive care units but its use its linked to a lot of complications. The objective of this study was to determine the risk factors for the presence of complications of the MVA in the newborns infants. MATERIAL AND METHODS: One hundred thirty five medical records from January 98 to June 2000 of newborns that had been discharged by amelioration or death were reviewed retrospectively. All of them had been received MVA; twenty medical record were excluded. They were divided in two groups: group A, neonates that had presented complications during MVA, composed of 40 patients (cases) and B, neonates with MVA, but that had not presented complications due to the procedure of 75 patients (controls). In group A there were 39 preterm neonates and in B, 58 preterm neonates, the rest were term neonates. It was used descriptive and inferential statistics. Odds ratio (OR) and multivariate analysis were used to study risk factors with confidence interval (CI) of 95%. Statistical significance was considered at p < 0.05. RESULTS: There was significative difference in birth weight. Gestational age and Apgar score between the two groups in favor of group B. There was significative difference too in inspiratory oxygen fraction (IOF), the peak inspiratory pressure (PIP) and cycles in favor of group A (before complications occur). The significative risk factors in the bivariate analysis for the presence complications were: birth weight of 1,500 g or less, 36 weeks of gestational age or less, total time of ventilation of seven days or more , IOF of 1 (100%), cycles of 60 per minute or more and peak inspiratory pressure (PIP) of 30 cm of water or more in the first days of life (all with p < 0.05). The combination of prematurity plus respiratory distress syndrome (RDS) as admission diagnosis reached significance for the presence of complications with an OR of 7.34 and CI of 95% from 1.91 to 23.05, p = 0.001 and the multivariate analysis reached significance for the presence of complications: low birth weight, total time of ventilation of seven days or more, IOF of 1 (100%), cycles of 60 per minute or more, and the combination of prematurity plus RDS as admission diagnosis all with p < 0.05. The mortality was associated significatively to the complications with p < 0.009. The more frequent complications were pneumonia, pulmonary interstitial emphysema and bronchopulmonary dysplasia. CONCLUSIONS: We concluded that newborn patients critically ill, with low birth weight, low gestational age, with RDS, who need more ventilatory support will be more liable to have complications and, as consequence, the mortality will be increased. For this reason it is essential a close monitoring of this group of patients for not over supporting them in IOF, PIP and cycles, thus avoiding as far as possible, the complications of the MVA. PMID- 15791907 TI - [Centipede (Scolopendra sp.) envenomation in a rural village of semi-arid region from Falcon State, Venezuela]. AB - INTRODUCTION: Centipedes are invertebrate animals belonging to Phylum Arthropoda, Class Chilopoda, with a first pair of front legs that has been modified to form large, poisonous fangs connected to venomous glands. Likely, due to relatively benignity of the clinical manifestations, in Venezuela studies and research on centipede envenomation are virtually scarse. AIM: The objective of this work was to study, biological, clinical, epidemiological and treatment profiles of envenomation caused by centipedes from Rio Seco town, semi-arid area of Falcon State, Venezuelan north-western region. METHODS: Between August 2001 and July 2002, patients who referred centipede attack were attended in ambulatory emergency. These were clinically examined, and by means of anamnesis the site and time of bite were recorded. The centipede was collected for identification. Ringer solution and non steroid analgesic plus hidrocortisone (500 mg, one doses) were administered parenterally and intravenously, respectively. In one case with abscess, the patient received intravenously antibiotic therapy for 3 days plus orally for 4 days (500 mg every 6 hours). RESULTS: It was registered 17 cases bitten by Scolopendra sp., aged 3-52 years, 9 females and 8 males, with an estimated Incidence Density (ID) of 1.34/100 persons/year. The majority of the accidents prevailed in the dry season (100%), inside the home (70.59%), during the night (52.94%) and on the hands (29.41%). Most clinical manifestations were local: hyperthermia (100%), pain (64.7%) and paresthesias (52.94%). The majority of the patients resolved symptoms quickly between 2-24 hours after cessation of the treatment (analgesic + hydrocortisone). CONCLUSION: Due to be expected an ID between 1.28 x 10(-12) and 0.010 (X = 0.0021)/100 persons/year, thus envenomation rate at Rio Seco, Falcon State, Venezuela, can be considered of "very high" or epidemic. Sinanthropic and domestic habits of scolopendromorphae centipedes increase the frequency of stings. Only symptomatic therapeutical treatment was required. PMID- 15791908 TI - [Cystic disease of the biliary tract. Three decades of institutional experence]. AB - INTRODUCTION: Cystic disease of biliary tract (CDBT) characterizes by the presence of sacular expansions of the biliary tree. It is an uncommon disease associated with high morbidity and malignant transformation. More than 60% of patients are women and can be diagnosed in the adult life. OBJECTIVE: To evaluate the results obtained during last three decades in the management of CDBT in the adult patient. PATIENTS AND METHODS: All the patients with CDBT treated from 1970 to 2002 were included. Demographic data, clinical picture, boarding diagnosis, classification, treatment, evolution and survival were analyzed. RESULTS: 34 patients. Twenty eight (82%) women and 6 (18%) men with a mean age of 33 years (range 13-84). The most frequent symptoms were abdominal pain, nausea-vomit and jaundice. Cholangiography was made in all cases. All the types described by Todani were documented. Twenty-seven patients (80%) were surgically treated. The mean follow-up was 84 months (range 1-408 months). Fifteen patients (44.1%) were readmitted and 9 (26.4%) had a reoperation. Three (9%) died with malignant transformation. The global survival was 91.1% to 12 months. CONCLUSIONS: In the adult patient, diagnosis of CDBT requires a high level of suspicion and its confirmation depends on the image studies. The CDBT diagnosis considers an indication of surgical treatment. Complete resection of the biliary tract with Roux en-Y hepato-jejunal anastomosis have less rate of mechanical complications, hospitalary readmissions and surgical reintervention. PMID- 15791909 TI - [Discrepancy in attitudes toward diabetes between physicians and diabetic patients attending the private, social security or state medical services]. AB - OBJECTIVE: One of the biggest challenges of the Mexican Health System is to get results of the treatment goals in diabetic patients, which can be frustrated by physician and patient's attitudes. The present study evaluates divergence of attitude of health providers and diabetic patients according to health sectors: private, social security or State medical services. MATERIAL AND METHODS: They contributed 255 physicians and 255 diabetic patients in ambulatory care from three health sectors: private, social security and welfare (employees and patients were exclusive providers and patients of corresponding institutions). The Diabetes Attitude Scale 3 (DAS-3) was applied, a self-administered instrument used to compare attitudes between health care professionals and patients. Discrepancies were analyzed with t-tests, chi2, one-way ANOVA and post hoc multiple comparisons for observed means based on the Tukey test. RESULTS: Physician and patient's attitudes differed in all scales except for psychosocial impact of diabetes, which was independent of the health sector. Attitudes were not alike with respect to severity of the disease, value of tight control of glucose levels and patient autonomy, with no distinction of health sector. Discrepancy of need for special training was the only attitude that varied by health sector (F = 4.1; p = 0.02); it was similar for the private and social security sector but different for the welfare sector. CONCLUSIONS: All health sectors should recognize attitude discrepancies between doctors and patients to favor collaborative strategies that benefit a better control of the disease. PMID- 15791910 TI - Comparison of available oxygen before and after red blood cell transfusion in critically-ill premature newborn infants of 28-36 weeks gestational age, to asses its possible usefulness in clinical practice. AB - BACKGROUND: Wardrop's formula to estimate available oxygen has been used to assess non-critically ill pre-term newborn infants (PTNIs) of 28-32 weeks gestational age who may need red blood cell transfusion ( RBCT ). The use of this formula has not been analyzed in critically-ill PTNIs. The objective of this study was to compare available oxygen levels before and after RBCTs in critically ill PTNIs of 28-36 weeks gestational age, and to assess is potential usefulness. MATERIAL AND METHODS: 113 clinical charts of critically-ill PTNIs with at least one RBCT performed between January 1998 and August 2001 were retrospectively reviewed. Hemoglobin concentrations, available oxygen levels and clinical signs of anemia (tachycardia, polypnea, pallor, failure to gain weight) were measured 3 days before, at the moment and 24-72 hours after the RBCT. Descriptive and inferential statistics were used to analyze the results. Statistical significance was considered when p < 0.05. RESULTS: Mean available oxygen levels were 7.99 +/- 1 mL/dL at the moment of the first RBCT (n = 113), 7.91 +/- 0.91 mL/dL at the second (n = 48), and 8.14 +/- 0.83 mL/dL at the third RBCT (n = 22). Available oxygen was significantly higher 24-72 hours after than before all RBCTs (p < 0.01). The mean available oxygen value at the moment of the first, second and third RBCTs was always near 8 mL/dL. CONCLUSIONS: Available oxygen is a good parameter to assess the need for RBCTs in critically-ill PTNIs of 28-36 weeks gestational age. Available oxygen values < 8mL/dL may be another parameter to consider as a criterion for RBCT in these patients. PMID- 15791911 TI - [A tetracycline-inducible gene system allows for regulated production of proteins with therapeutic potential]. AB - The adoption of a bacterial system of control of genic expression with tetracycline, combined with the advances in the identification of regulatory sequences and mechanisms of expression of eukaryotic genes, has increased the versatility and effectiveness of techniques to reintroduce and to make genes function in cells and in superior organisms by being able to control the activity of the transgene in a temporary or reversible manner. This approach has also facilitated making detailed studies of different cellular processes and diseases; for example, the study of the function of oncogenes and other genes involved in the formation and progression of cancer, the generation of cellular models for recombinant protein production with therapeutic purposes, the ex-vivo genetic manipulation of extracted cells from patients and returned to them for gene therapy procedures, as well as the modulation of transgenes to revert hereditary suffering in animal models. PMID- 15791912 TI - [Free amino acids in the amniotic fluid and maternal plasma in different periods of the normal pregnancy]. AB - OBJECTIVE: To describe changes in the concentration of total free amino acids (TFAA) in amniotic fluid (AF) and in maternal plasma (MP) at different periods of pregnancy, to analyze their differences, and to correlate the composition of the aminograms (AMG). METHODS: A total of 19 free AA. were measured by chromatography in the AF and MP from 18 women with uncomplicated singleton pregnancies, 6 at 16 weeks of gestation (wg), 6 at 28 wg and, 6 at 36 wg. Differences in the TFAA concentration at each gestational age, essential/non-essential, and polar/non polar AA ratios were calculated. The correlation between the AMG in AF and in MP at each gestational age, was also estimated. RESULTS: In the AF there was a reduction in the concentration of TFAA throughout pregnancy (p = 0.023) whereas, in MP the TFAA concentration did not change. TFAA concentration was lower in AF than in MP at 36 weeks of gestation (p = 0.001). The composition of the AMG in MP did not change during pregnancy with a correlation coefficient between 16 and 28 wg of r = 0.98 (95% confidence interval [95%CI] 0.94-0.99), between 16 and 36 wg: r = 0.95 (95% CI 0.86-0.99) and between 28 and 36 wg: r = 0.93 (95% CI 0.82 0.97). The ratio of essential/non-essential and polar/non-polar AA was also unchanged. In AF, the composition of the AMG changed during pregnancy with a correlation between 16 and 28 wg: r = 0.83 (95% CI 0.59-0.93), and between 16 and 36 wg: r = 0.78 (95% CI 0.49-0.91). This was due to a reduction in the basic AA and an increment in the acid and neutral AA. The correlation between the AMG in AF and in MP at 16 wg was: r = 0.94 (95% CI 0.84-0.96), at 28 wg: r = 0.82 (95% CI 0.57-0.92) and, at 36 wg: r = 0.71 (95% CI 95% 0.37-0.88). CONCLUSION: During pregnancy, TFAA and the AMG in MP remained stable. In AF, TFAA were reduced and the AMG changed towards the end of the pregnancy. PMID- 15791914 TI - [Prostate cancer]. PMID- 15791913 TI - [Insulin resistance inpolycystic ovary syndrome]. PMID- 15791915 TI - [Evolution of knowledge on blood and its movement. 1st Part. Integration of circulatory doctrine. Iatrophysics of blood]. AB - Under the light of the evolution of human knowledge, we present a short historic outline of the integrating steps of the blood circulation doctrine and its components, a starting point for ulterior studies on the function of blood in the economy of an organism. The different steps of the process leading to the integration and diffusion of the blood circulation doctrine are described. These go from the first descriptions of the heart structures by authors of the ancient world to the careful anatomical observations of Renaissance scientists, such as Leonardo da Vinci. Towards the middle of the sixteenth century, the first well known descriptions of pulmonary circulation were made independently by Miguel Servet and Realdo Colombo. Later, William Harvey, based on the experimental quantitative method according to Galileo's thought, described and studied systemic circulation. The vicissitudes encountered by the circulation doctrine are also disclosed. At the beginning, this concept was rejected by the traditional milieu--such as the Sorbonne of Paris--, but was accepted by the young and innovative researchers in both England and Continental Europe. In the second half of the seventeenth century and the first half of the eighteenth, the blood circulation doctrine was finally accepted and spread throughout Europe and the American continent. Furthermore, this doctrine inspired the medical practice of intravenous injections and blood transfusions. Its evolution, as generally occurs in the evolution of scientific thinking, allowed to reject some believes deeply rooted in the doctrine's constituent structure, i.e., the Renaissance spell of circularity. PMID- 15791916 TI - Developmental screening. AB - An estimated 5 to 10% of the pediatric population has a developmental disability. The current strategy to identify these children is through developmental surveillance, a continuous procedure in which the health professional observes the infant, takes a developmental history, and elicits any concerns that the caregiver might have. However, identification of delayed children is ineffective when based solely on routine surveillance. A necessary adjunct is developmental screening: the process of systematically identifying children with suspected delay who need further assessment. Screening tests greatly improve the rate of identification. With the advent of intervention programs and the support of organizations such as the American Academy of Pediatrics, the topic of developmental screening is a timely and essential one. This review aims to describe the properties of screening tests, to evaluate the available tools for developmental screening while providing a representative sample of the currently available developmental tests, and, finally, to evaluate the efficacy of intervention programs, a needed prerequisite to justify screening. PMID- 15791917 TI - Melatonin excretion in normal children and in tuberous sclerosis complex with sleep disorder responsive to melatonin. AB - To determine normal melatonin excretion patterns in healthy children without sleep disorder and to compare these with those of patients with tuberous sclerosis complex and sleep disorder responsive to exogenous melatonin, we measured 6-sulfatoxymelatonin excretion in 21 healthy children and in 7 patients with tuberous sclerosis complex and sleep disorder responsive to melatonin (a 5 mg oral dose increasing total sleep time). Total excretion, cosinor percentage, and acrophase time of 6-sulfatoxymelatonin excretion were estimated. In normal children, total 6-sulfatoxymelatonin excretion was range 11.1 to 40.2 microg (mean 19.0 microg, SD 7.4 microg); cosinor percentage rhythm range was 52.9% to 100% (mean 87%, median 94%); and acrophase time range was 23 hours, 54 minutes to 10 hours, 42 minutes (mean 5 hours, 54 minutes; median 4 hours, 12 minutes). Fifth and 95th percentiles were 11.1 to 29.0 microg, 57.8% to 99.9%, and 2 hours, 1 minute to 10 hours, 4 minutes. In tuberous sclerosis, normal patterns of melatonin excretion were seen in responders. Circadian patterns of melatonin excretion were similar in children and adults. We propose that exogenous melatonin can act by a simple sedative action. PMID- 15791918 TI - Board certification in child neurology and neurology: cohort study. AB - This study followed the progress of a cohort of child neurology and neurology graduates who sought certification by the American Board of Psychiatry and Neurology (ABPN). The 211 candidates were recent graduates when they initiated the certification process in October 1994, and 92% of them have achieved certification. First-attempt performances on the Part I and Part II examinations were related; 62% of those certified passed both on their first attempts. Of those who were certified, 24% also achieved certification in at least one additional ABPN specialty (psychiatry) or subspecialty (clinical neurophysiology, neurodevelopmental disabilities, and pain medicine). Overall, it was estimated that 82% of the 1994 child neurology and neurology graduates sought ABPN certification, and of these, 82% were successful. PMID- 15791919 TI - Epilepsy, electroencephalographic abnormalities, and regression in children with autism. AB - The association of epilepsy and autism is recognized, and it has been reported at a percentage that varies between 8 and 42%, depending on age and diagnostic criteria. One third of autistic children undergo a regression of language and behavior between 2 and 3 years, and epileptiform abnormalities and epilepsy can be concomitant in an undetermined percentage of them. The aim of this study was to investigate the prevalence of epilepsy and paroxysmal abnormalities in a group of children with autism and to determine the percentage of regression course in this group. Forty-six patients with autism (mean age 7.8 +/- 2.7 years; 34 boys and 12 girls) were consecutively examined, and clinical evaluation, assessment, and electroencephalographic (EEG) recordings were performed in all of them. Thirty-five percent showed paroxysmal abnormalities and epilepsy, 22% had only paroxysmal abnormalities without seizures, and 13% of the children suffered from epilepsy. Sixty-five percent had a normal EEG. No difference in regression rate was observed between patients with paroxysmal abnormalities and epilepsy and those with a normal EEG and without seizures. In the study group, the prevalence of epilepsy was in the low range of individuals with autism, and different types of epilepsy were observed. Autism with regression was not influenced by paroxysmal abnormalities and epilepsy. PMID- 15791920 TI - Experience in the use of the ketogenic diet as early therapy. AB - The ketogenic diet has traditionally been considered an anticonvulsant therapy of last resort, despite excellent efficacy and limited side effects. We hypothesized that the ketogenic diet would have similar results in patients with new-onset epilepsy. A retrospective study was conducted of patients started on the ketogenic diet since 1994. Thirteen of 460 (2.8%) patients were started on the ketogenic diet as early (zero or one prior anticonvulsant) therapy for seizures. Of those remaining on the diet, 60% (6 of 10) had a > 90% seizure reduction at 6 months and 100% (6 of 6) had a > 90% reduction at 12 months. Patients with infantile spasms were as likely to achieve > 50% seizure reduction at 6 months as patients with other seizure types (75% vs 60%; P = .6). The ketogenic diet can be a valuable therapy before epilepsy becomes intractable. In the 13 patients reported, efficacy without side effects was achieved similarly to that with patients with intractable epilepsy. PMID- 15791921 TI - Macrocephaly, corpus callosum morphology, and autism. AB - Although the cause of autism is undetermined, a general consensus has been that some type of early aberrant neural development underlies the disorder. Given the increased prevalence of macrocephaly in autism, one theory of abnormal neural development implicates early brain growth resulting in larger brain and head size in autism. Surface area measurements of the midsagittal section of the corpus callosum can be used as an index of neural development and white-matter integrity because the corpus callosum is the major white-matter structure that interconnects the two cerebral hemispheres. The purpose of this study was to obtain corpus callosum surface area, shape, and contour in a sample of non mentally retarded autistic subjects with macrocephaly (n = 12) and compare them with those of matched (n = 8), typically developing control subjects with benign macrocephaly. No significant differences were found in surface area, shape, or contour between groups, nor did corpus callosum surface area relate to measures of IQ or picture vocabulary. These findings suggest no unique difference in overall regional corpus callosum surface area in autism with macrocephaly. PMID- 15791922 TI - Health service use in Rett syndrome. AB - Rett syndrome is a severe neurodevelopmental disorder affecting girls, caused by mutations in the MECP2 gene. There are no population-based data on the extent and determinants of health service use in this disorder. The population-based registry, the Australian Rett Syndrome database, was the source of phenotype data. This also contains data from mutation screening and X-inactivation studies. Data on retrospective (n = 152) and prospective (n = 162) health service use were collected in 2000 from a questionnaire and a calendar study, respectively. Health service use was highest in younger cases (P = .001) and lowest in cases with milder phenotypes (P < .001). Random X-inactivation was associated with service use (P = .02). Maternal education, phenotype, and individual mutations were determinants of health service use. The use of a retrospective and prospective data set enabled accurate assessment of service use in Rett syndrome. Both genetic and sociodemographic determinants of health service use were identified, with important implications for the optimal and equitable management of children with Rett syndrome. PMID- 15791923 TI - Neuroprotective effects of brain-derived neurotrophic factor (BDNF) on hearing in experimental pneumococcal meningitis. AB - Bacterial meningitis is still one of the most common causes of acquired profound sensorineural deafness in children despite antibiotic treatment. We investigated the neuroprotective effects of brain-derived neurotrophic factor on hearing function in experimental bacterial meningitis. We implanted stainless steel tubes into both cerebral ventricles of Sprague-Dawley rats aged 21 days. Bacterial meningitis was induced by inoculating a strain of serotype III Streptococcus pneumoniae into the cisterna magna. Six micrograms per day of brain-derived neurotrophic factor (groups 1 and 3) or albumin (groups 2 and 4) was injected into the cerebral ventricles 24 hours after or before infection, respectively, for a duration of 7 days. Additionally, all rats received antibiotic subcutaneous treatment starting 24 hours after infection for 7 days. Brainstem auditory evoked potentials were recorded 24 hours before and 24 hours after infection and after 7 days of treatment with brain-derived neurotrophic factor or placebo and antibiotics, respectively, to determine hearing threshold. Our results showed that the hearing thresholds of animals in each group increased significantly 24 hours after infection compared with the results recorded 24 hours before infection (P < .01). After 7 days of treatment with brain-derived neurotrophic factor, brainstem auditory evoked potential responses recurred in 16 ears when stimulated at 75 dB hearing level in groups 1 and 3. Their hearing thresholds significantly decreased compared with the control group 2 (P < .05) and group 4 (P < .01). However, 13 of 14 ears absent brainstem auditory evoked potential responses could still not be identified at 75 dB hearing level in control groups 2 and 4. The improvement of the hearing thresholds in group 3 (treated before infection) was greater than that of group 1 (treated after infection) (P < .05), but there was no significant difference found between the control groups before and after infection (P > .05). Our study supports the hypothesis that the administration of exogenous brain-derived neurotrophic factor can be effective in preventing or treating hearing loss following bacterial meningitis. PMID- 15791924 TI - Magnetic resonance imaging findings and novel mutations in GM1 gangliosidosis. AB - Two unrelated children and their siblings of Arab origin were diagnosed as having GM1 gangliosidosis on the basis of clinical features and markedly low levels of beta-galactosidase. The T2-weighted magnetic resonance images of the brain revealed certain characteristic features, including delayed myelination and abnormal appearance of the subcortical white matter, internal capsule, and basal ganglia. Their mutation analysis showed two novel mutations, which have not been described in an Arabic population. PMID- 15791925 TI - Frequency and nature of cerebellar injury in the extremely premature survivor with cerebral palsy. AB - We report on the frequency and variable nature of magnetic resonance imaging documented injury to the cerebellum in children with cerebral palsy who had survived a birth with a weight under 1000 g and/or a gestational age under 28 weeks. Thirty of 67 patients who had magnetic resonance images were found to have injury to the cerebellum. Those with cerebellar injury were much more likely to be microcephalic and to be unable to walk or talk. They did not demonstrate a greater frequency of observed injury to the cerebrum. From a larger collection of children with known cerebellar injury and cerebral palsy who had a history of being extremely premature, we found that 35 of 47 patients had prominent injury to the inferior cerebellum, suggesting infarction, whereas the remainder demonstrated varying degrees of cerebellar atrophy with or without asymmetry and four also had enlarged 4th ventricles. Injury to the cerebellum in the extremely premature survivor who has cerebral palsy is common and associated with a more adverse clinical picture. The etiology of this injury is obscure. PMID- 15791926 TI - Residual motor control and cortical representations of function following hemispherectomy: effects of etiology. AB - Fifteen posthemispherectomy children were examined to assess residual motor function of the paretic side using the 74-point Fugl-Meyer Assessment of Motor Recovery scale. The degree of residual motor control differed for upper and lower extremities, with hand function being most severely impaired. Posthemispherectomy motor outcomes also differed as a function of etiology: cortical dysplasia, perinatal infarct, and Rasmussen's encephalitis. Children whose intractable seizures resulted from perinatal middle cerebral artery stroke demonstrated the most spared motor function. To detect cortical areas that represented motor control of the hemiparetic side, we focused on voluntary control of the affected lower extremity. Seven of our patients were able to carry out a foot dorsiflexion paradigm during functional magnetic resonance imaging, and these results were compared with activations found in normal controls. All children showed activations in the sensorimotor network ipsilateral to the affected side. The perinatal infarct group demonstrated greater activity in the cingulate cortex, whereas the Rasmussen's encephalitis group had significant activations in the insula, suggesting etiology-specific differences in reorganization. These findings are discussed in the framework of our understanding of mechanisms of cortical plasticity in the injured brain and its relevance to neurologic rehabilitation. We suggest that imaging techniques are important tools in identifying cortical regions underlying functional reorganization. Furthermore, detection of such areas might become a basis for specific training promoting the optimal reorganization of cortical networks to enhance motor control. PMID- 15791927 TI - Neurogenic arthrogryposis multiplex congenita and velopharyngeal incompetence associated with chromosome 22q11.2 deletion. AB - We report a case of neurogenic arthrogryposis multiplex congenita and velopharyngeal incompetence in association with a chromosome 22q11.2 deletion in a 5-month-old boy, the only child of a non-consanguineous couple without relevant antecedents. Specifically, polymerase chain reaction amplification of microsatellite markers revealed a noninherited microdeletion in position D22S306. This phenotype has not been reported previously in association with chromosome 22q11.2 deletions, and these findings raise the possibility that at least some cases of neurogenic arthrogryposis multiplex congenita might be due to genetic defects of this type. PMID- 15791929 TI - Two siblings with familial idiopathic scoliosis with conjugate gaze palsy. AB - Familial idiopathic scoliosis with conjugate gaze palsy is a rare autosomal recessive disorder characterized by progressive scoliosis and congenital absence of conjugate horizontal eye movement. In this article, two siblings who had scoliosis and conjugate gaze palsy are presented regarding clinical and laboratory findings. PMID- 15791928 TI - Effect of melatonin dosage on sleep disorder in tuberous sclerosis complex. AB - We report a randomized, double-blind, controlled, crossover trial investigating the response to oral melatonin using two dose regimens in patients with sleep disorders associated with tuberous sclerosis complex. Eight outpatients with tuberous sclerosis complex and sleep disorder received either 5 or 10 mg of melatonin. Sleep latency, total sleep time, number of awakenings, and seizure frequency were recorded in sleep and seizure diaries. No evidence of a dose effect between 5 and 10 mg was seen with respect to any outcome measure. (The 5 mg results are given first: sleep latency, 86 and 76 minutes; total sleep time, 8 hours, 57 minutes and 9 hours, 4 minutes; and sleep fragmentation, 0.8 and 1.0). This study might have missed a small beneficial effect of 10 mg melatonin. We propose that an initial trial of 5 mg melatonin is worth considering in patients with tuberous sclerosis complex and sleep disorder. PMID- 15791930 TI - Four-month-old infant with focal segmental glomerulosclerosis and mitochondrial DNA deletion. AB - Mitochondrial cytopathies are a group of heterogeneous disorders characterized by multisystem involvement. Renal involvement in mitochondrial cytopathies is usually manifested as tubular dysfunction owing to impaired energy metabolism; however, a few cases with glomerular changes have also been reported. Herein we report the case of a 4-month-old Turkish girl with a mitochondrial DNA deletion and focal segmental glomerulosclerosis. PMID- 15791931 TI - Extraction of primary and secondary metabolites. AB - Several new methods besides the usual organic solvent extraction have been developed over the last few years for the extraction of primary and secondary metabolites. These are: alcohol extraction with various biocompatible solvents, recovery of carboxylic acids and antibiotics with reactive extraction, dissociation extraction, aqueous two-phase extraction, and supercritical and near critical fluid extraction. Extraction and re-extraction processes are integrated into a single step by emulsion liquid membrane and solid supported liquid membrane extractions. These extraction processes are discussed and compared in this review, along with extraction with reversed micelles, and reactive extraction with the formation of a third phase at the organic-aqueous interface. PMID- 15791932 TI - Biochemical reaction engineering and process development in anaerobic wastewater treatment. AB - Developments in production technology have frequently resulted in the concentrated local accumulation of highly organic-laden wastewaters. Anaerobic wastewater treatment, in industrial applications, constitutes an advanced method of synthesis by which inexpensive substrates are converted into valuable disproportionate products. A critical discussion of certain fundamental principles of biochemical reaction engineering relevant to the anaerobic mode of operation is made here, with special emphasis on the roles of thermodynamics, kinetics, mass and heat transfer, reactor design, biomass retention and recycling. The applications of the anaerobic processes are discussed, introducing the principles of an upflow anaerobic sludge bed reactor and a fixed-bed loop reactor. The merits of staging reactor systems are presented using selected examples based on two decades of research in the field of anaerobic fermentation and wastewater treatment at the Forschungszentrum Julich (Julich Research Center, Germany). Wastewater treatment is an industrial process associated with one of the largest levels of mass throughput known, and for this reason it provides a major impetus to further developments in bioprocess technology in general. PMID- 15791933 TI - The oxygen mass transfer, carbon dioxide inhibition, heat removal, and the energy and cost efficiencies of high pressure fermentation. AB - This study focuses on the enhancement of oxygen transfer in aerobic fermentation processes through the use of the reactor headspace pressure as a process variable. Our investigation was performed using an example 5 m3 fermentor, and by applying the mass transfer (sorption) characteristics obtained from measurements with Pseudomonas putida CA-3 in a mineral medium. By applying reactor pressures up to 11 bar (10 bar overpressure) the oxygen transfer capacities of stirred tank reactors can theoretically be increased to values above 1 kmol m(-3) h(-1), although the problem of heat removal limits that value to levels of 0.7 kmol m( 3) h(-1). However, this is still sufficient for the oxygenation of most aerobic fermentation processes, even at very high cell densities. Possible inhibition by the carbon dioxide partial pressure at elevated total pressures is discussed. It is shown that increasing reactor pressure enhances the energy efficiency, defined as the ratio of the oxygen transfer capacity to the total power consumption of the reactor system. The cost efficiency of the oxygen mass transfer, defined as the ratio of the oxygen transfer capacity to the overall cost of the system (overall purchase cost divided by a project lifetime of ten years, plus total energy cost) also increases with reactor pressure up to 5.6 bar. PMID- 15791934 TI - Biochemical engineering aspects of expanded bed adsorption. AB - The economic feasibility of a biotechnological production process is directly linked to the number and efficiency of the processes required during purification in order to reach the requested product specifications. Expanded bed adsorption is an integrated process that combines solid-liquid separation and product recovery into a single unit operation. This approach suggests an increased overall yield, lower requirements for capital investments and consumables and most importantly a reduced process time. The success of an integrative process is, however, closely linked to a detailed understanding of the biochemical principles involved and the constraints arising from feedstock complexity. This paper reviews the hydrodynamic principles that expanded bed adsorption is based on, discusses current developments in resin and column design, and finally presents a methodology for targeted process development in expanded bed adsorption. PMID- 15791935 TI - Parallel reactor systems for bioprocess development. AB - Controlled parallel bioreactor systems allow fed-batch operation at early stages of process development. The characteristics of shaken bioreactors operated in parallel (shake flask, microtiter plate), sparged bioreactors (small-scale bubble column) and stirred bioreactors (stirred-tank, stirred column) are briefly summarized. Parallel fed-batch operation is achieved with an intermittent feeding and pH-control system for up to 16 bioreactors operated in parallel on a scale of 100 ml. Examples of the scale-up and scale-down of pH-controlled microbial fed batch processes demonstrate that controlled parallel reactor systems can result in more effective bioprocess development. Future developments are also outlined, including units of 48 parallel stirred-tank reactors with individual pH- and pO2 controls and automation as well as liquid handling system, operated on a scale of ml. PMID- 15791936 TI - From stationary to instationary metabolic flux analysis. AB - Metabolic flux analysis using 13C labeled substrates is an important tool for metabolic engineering. Although it has now been evolving for more than ten years, metabolic flux analysis has still not reached the limits of its application. Current developments aim at extending to instationary industrial production conditions like batch and fed-batch fermentations on the one hand, and at miniaturization and high throughput flux analysis on the other. In both cases, reducing the duration of the labeling experiment is of major interest. For that reason, there is now interest in the instationary transient of the 13C enrichment during a labeling experiment. This paper presents some recent developments in the field of instationary metabolic flux analysis and discusses some critical aspects and limitations using some simulation examples. PMID- 15791937 TI - Applying metabolic profiling techniques for stimulus-response experiments: chances and pitfalls. AB - So far it is mainly transcriptome and proteome analysis that has been applied to elucidate the correlation between genotype and phenotype although thorough metabolome studies can provide substantial information on the control of the metabolism at the biochemical level. Stimulus-response experiments, i.e. the investigation of metabolism dynamics after a glucose pulse (pulse experiment), can be used to study the in vivo enzyme kinetics offering insight into underlying reaction mechanisms. Usually, this requires rapid cell quenching combined with cell inactivation to'freeze' the microbial metabolism response at a definite time lag after pulse stimulation. To access the 'frozen' metabolic reply, adequate analytical methods are needed to measure intracellular metabolite concentrations in the cell extract. As shown in the introductory review part, stimulus-response experiments were usually applied to study central metabolism dynamics in wildtype strains. Our own results, presented in the second part of the contribution, indicate that stimulus-response experiments should also be applied to analyse pathway dynamics in anabolic routes. Using the example of the aromatic amino acid pathway, an LC-MS/MS technique is presented that allows the quantification of intracellular pools of central metabolism as well as of the aromatic amino acid pathway. Based on the analytical approach metabolic profiling is performed to monitor the metabolism dynamics after a glucose pulse experiment allowing the conclusion that pulse stimulation is transmitted to the anabolic pathway of interest. PMID- 15791938 TI - Technical application of biological principles in asymmetric catalysis. AB - The production of enantiopure compounds is becoming increasingly important to the chemical and biotechnological industries. Bioorganic transformations look set to meet this demand due to their inherently regio- and stereoselective natures. In this sense, biosynthesis needs to be viewed as "chemistry by nature". Biological principles that have been optimized over thousands of years experience a new renaissance when used for technical asymmetric catalysis; however, to be able to use them, we need an appropriate technology: reaction engineering. Indeed, various biological principles are already being applied in technical asymmetric synthesis without the scientific community at large being aware of this. PMID- 15791939 TI - Cofactor regeneration at the lab scale. AB - Progress made in lab-scale applications of various coenzyme regeneration systems over the last two decades has mainly focused on the applications of NAD+/NADH- and NADP+/NADPH-dependent oxidoreductase reactions. In situ regeneration systems for these reactions, as well as whole cell, enzymatic, electro-enzymatic, chemical, and photochemical reactions are presented, including details about their efficiency and novelty. The progress of enzyme reaction engineering is also reported. PMID- 15791940 TI - Enzyme-catalyzed regio- and enantioselective ketone reductions. AB - In this chapter, examples are given of the application of highly reactive prochiral ketones as substrates for enzymatic reductions. 3,5-Dioxocarboxylates are polyketide-like compounds that can be used to synthesize all of the possible stereoisomers of the corresponding 1,3-diols by means of regio- and enantioselective enzymatic reduction. The results obtained from an investigation into the usefulness of the resulting hydroxyl ketones and 1,3-diols in organic synthesis led to the development of non-natural functionalized ynones as a starting material for the enzymatic route to enantiopure propargylic alcohols. A broad variety of substituted acetylenic ketones can be reduced enantioselectively by the oxidoreductases Lactobacillus brevis ADH (LBADH), Candida parapsilosis carbonyl reductase (CPCR), horse liver ADH (HLADH) and Thermoanaerobium brockii ADH (TBADH). The resulting propargylic alcohols can be obtained in either enantiomeric form, since (R)- and (S)-specific oxidoreductases can be applied. By varying the size of the substituents, the enantiomeric excess can be tuned, or the enantioselectivity can even be reversed. The obtained highly functionalized enantiopure alcohols are synthetically flexible chiral building blocks that offer new synthetic strategies for target- and diversity-oriented synthesis. PMID- 15791941 TI - Membrane reactors at Degussa. AB - The review covers the development of membrane reactor technologies at Degussa for the synthesis of fine chemicals. The operation of fed-batch or continuous biocatalytic processes in the enzyme membrane reactor (EMR) is well established at Degussa. Degussa has experience of running EMRs from laboratory gram scale up to a production scale of several hundreds of tons per year. The transfer of the enzyme membrane reactor from biocatalysis to chemical catalysis in the chemzyme membrane reactor (CMR) is discussed. Various homogeneous catalysts have been investigated in the CMR, and the scope and limitation of this new technique is discussed. PMID- 15791942 TI - Facts and theories of induced organ regeneration. AB - Induced organ regeneration is de novo synthesis of a physiological, or nearly physiological, organ at the same anatomical site as the organ that is being replaced. Regeneration of skin, peripheral nerves and the conjunctiva, described in this chapter, have been accomplished using biologically active scaffolds (regeneration templates) seeded with epithelial cells; devices for regeneration of the first two organs are in clinical use. There is substantial empirical evidence that templates induce regeneration by blocking contraction, the major mechanism for closure of severe wounds in adults. Templates appear to function by interfering with normal myofibroblast function well as by acting as temporary configurational guides for synthesis of new stroma that resembles that of the organ under replacement. The combined evidence supports a theory which predicts that selective blocking of the adult healing response uncovers the latent fetal response to injury and leads to organ regeneration. An independent theory suggests that loss of regenerative potential during the mammalian fetal-adult transition is associated with simultaneous acquisition of individual immunocompetence. PMID- 15791943 TI - Regenerative capacity and the developing immune system. AB - Many components of the vertebrate immune system have evolved with dual, interrelated functions of both protecting injured tissues from infection and providing for tissue maintenance and repair of injuries. The capacity for organ regeneration, prominent among invertebrates and certain phylogenically primitive vertebrates, is poorly developed in mammals. We have proposed that evolution of the mammalian immune system has produced inflammatory cellular interactions at sites of injury which have optimized tissue defense and facilitated tissue repair, but that these improvements included concomitant loss of regenerative capacity. This chapter briefly reviews work in two regenerating systems: scar free repair of fetal mammalian skin and regeneration of amputated limbs in larval frogs. In both organs the potential to regenerate anatomically and functionally complete new structures is lost gradually during ontogeny and this loss coincides with development of an immune system producing an inflammatory response in injured tissues. Failure of organ regeneration has long been associated with scarring or fibrosis and this phenomenon is a direct result of inflammatory interactions of immune cells and fibroblasts at sites of injury. Several aspects of immunity related to repair are reviewed, including the importance of antigen presenting cells and lymphocytes, relevant cytokines and growth factors released by these and other cells, immune functions of extracellular matrix components, and immunological functions of fibroblasts. Skin repair in various transgenic mouse models has been especially informative. Further study of immune mechanisms associated with the loss of regenerative capacity in the skin and amphibian limb will be useful for efforts to promote mammalian organ regeneration. PMID- 15791944 TI - Vertebrate limb regeneration. AB - In this chapter, we have touched upon some of the key processes of vertebrate limb regeneration from the formation of the wound epithelium to pattern formation, to provide a picture of the many complex and intricate facets of this system. Our synthesis incorporates recent advances in molecular biology, which has revealed some important factors related to the initiation, induction and patterning in limb regeneration. PMID- 15791945 TI - Mammalian fetal organ regeneration. AB - The developing fetus has the remarkable ability to heal dermal skin wounds by regenerating normal epidermis and dermis with restoration of the extracellular matrix architecture, strength, and function. The biology responsible for scarless wound healing in skin is a paradigm for ideal tissue repair. This regenerative capacity is lost in late gestation when fetal wounds heal with fibrosis and scar. Early in gestation, fetal skin is developing at a rapid pace in a unique environment. Investigation of normal skin embryogenesis and comparison between early scarless and late scarring fetal wounds has revealed distinct differences in inflammatory response, cellular mediators, wound contraction, cytokines, growth factors, and extracellular matrix modulators. The knowledge gained from comparative observational studies has served as a base for experimental interventions in animal models to induce or ameliorate scar. Although much progress has been made over the past decade, the mechanism of fetal wound healing remains largely unknown and attempts to mimic the scarless wound phenotype have not been completely successful. Identification of more key genes involved in skin regeneration may have implications in adult skin wounds and repair in other organ systems. PMID- 15791946 TI - Liver regeneration. AB - This review summarizes the functional aspects, cellular kinetics and molecular mechanisms related to liver regeneration. Liver regeneration is a model of regenerative growth of tissues in adult animals. Rapid biochemical and gene expression changes following initiation of regeneration are mediated by specific stimuli, including growth factors and cytokines. The whole process involves multiple feedback loops between same or different types of cells. The end result is restoration of hepatic mass and preservation of the normal histology of the liver. All of this complex phenomenology occurs while the liver continues to provide all the metabolic support required to sustain life of the organism. PMID- 15791947 TI - Stem cells in CNS and cardiac regeneration. AB - The central nervous system (CNS) and the heart muscle regenerate poorly after injury, yet evidence is mounting that both harbor cells capable of rebuilding neural and cardiac tissue. The reason for the poor regenerative response of CNS tissue and myocardium must therefore lie in the nature of the injury environment, which promotes fibrosis over regeneration. Strategies for regenerating these tissues thus rely on overcoming the fibrotic response by filling lesions with tissue-specific regeneration-competent cells that replace or rescue dying cells, or by activating endogenous regeneration-competent cells that do likewise. There has also been considerable excitement about the possibility of transplanting bone marrow cells into CNS or cardiac lesions to repair them, because bone marrow cells have been reported to be pluripotent. In this chapter, contemporary evidence for the existence of regeneration-competent cells in the CNS and heart is discussed, as well as attempts to use these cells and bone marrow cells to reconstitute new tissue. PMID- 15791948 TI - Island grafts: a model for studying skin regeneration in isolation from other processes. AB - This chapter introduces the concept of an island graft, in which large full thickness skin wounds are created with a small treatment area in the center. The island graft is a simple method for demonstrating the effect of epithelial derivatives on wound regeneration without using sophisticated biologic or genetic markers. In the absence of disease, mammalian wounds of the dermis close by scarring and contraction. The island graft permits observation of the test material within the wound without interference from the periphery for several days until wound contraction and epithelialization of the surrounding skin encroach upon the study area. PMID- 15791950 TI - Poliovirus, pathogenesis of poliomyelitis, and apoptosis. AB - Poliovirus (PV) is the causal agent of paralytic poliomyelitis, an acute disease of the central nervous system (CNS) resulting in flaccid paralysis. The development of new animal and cell models has allowed the key steps of the pathogenesis of poliomyelitis to be investigated at the molecular level. In particular, it has been shown that PV-induced apoptosis is an important component of the tissue injury in the CNS of infected mice, which leads to paralysis. In this review the molecular biology of PV and the pathogenesis of poliomyelitis are briefly described, and then several models of PV-induced apoptosis are considered; the role of the cellular receptor of PV, CD155, in the modulation of apoptosis is also addressed. PMID- 15791949 TI - Mechanisms of apoptosis during reovirus infection. AB - Reovirus infection has proven to be an excellent experimental system for studying mechanisms of virus-induced pathogenesis. Reoviruses induce apoptosis in a wide variety of cultured cells in vitro and in target tissues in vivo, including the heart and central nervous system. In vivo, viral infection, tissue injury, and apoptosis colocalize, suggesting that apoptosis is a critical mechanism by which disease is triggered in the host. This review examines the mechanisms of reovirus induced apoptosis and investigates the possibility that inhibition of apoptosis may provide a novel strategy for limiting virus-induced tissue damage following infection. PMID- 15791951 TI - Neuronal cell death in alphavirus encephalomyelitis. AB - Alphaviruses are mosquito-borne, enveloped, plus-strand RNA viruses that cause a spectrum of diseases in humans that include fever, rash, arthritis, meningitis, and encephalomyelitis. Sindbis virus (SINV) is the prototype alphavirus, causes encephalomyelitis in mice, and provides a model system for studying the pathogenesis of alphavirus-induced neurological disease. Major target cells for SINV infection in the central nervous system (CNS) are neurons, and both host and viral factors determine the fate of infected neurons. Young animals are most susceptible to fatal disease. This correlates with the ability of SINV to induce apoptosis in immature neurons. In vitro, apoptotic death of neuroblastoma cells can be induced by fusion of the virus envelope with the endosomal membrane and does not require infectious virus. This fusion process activates acid sphingomyelinase that cleaves sphingomyelin to release ceramide, an initiator of apoptosis. Within an hour, poly(ADP-ribose) polymerase is activated, and this is followed by release of cytochrome c and activation of effector caspases. SINV induced cell death can be delayed or prevented by treatment with antioxidants or caspase inhibitors and by intracellular expression of Bcl-2, Beclin-1, or protease inhibitors. Older animals survive infection unless infected with a neurovirulent strain of SINV. In these mice, anterior horn motor neurons die by a primarily necrotic process that is influenced by excitotoxic amino acids and inflammation, whereas hippocampal neurons can be either apoptotic or necrotic. Death also occurs in uninfected neurons in the vicinity of infected neurons and can be delayed or prevented by treatment with glutamate receptor antagonists. PMID- 15791952 TI - HSV-induced apoptosis in herpes encephalitis. AB - HSV triggers and blocks apoptosis in cell type-specific fashion. This review discusses present understanding of the role of apoptosis and signaling cascades in neuronal pathogenesis and survival and summarizes present findings relating to the modulation of these strictly balanced processes by HSV infection. Underscored are the findings that HSV-1, but not HSV-2, triggers apoptosis in CNS neurons and causes encephalitis in adult subjects. Mechanisms responsible for the different outcomes of infection with the two HSV serotypes are described, including the contribution of viral antiapoptotic genes, notably the HSV-2 gene ICP10PK. Implications for the potential use of HSV vectors in future therapeutic developments are discussed. PMID- 15791953 TI - The role of apoptosis in defense against baculovirus infection in insects. AB - The baculoviruses make up a large, diverse family of DNA viruses that have evolved a number of fascinating mechanisms to manipulate their insect hosts. One of these is the ability to regulate apoptosis during infection by expressing proteins that can inhibit caspase activation and/or activity, including the caspase inhibitor P35 and its relatives, and the inhibitor of apoptosis (IAP) proteins. Experimental manipulations of the expression of these antiapoptotic genes, either by genetic deletions or by RNAi, have shed light on the effectiveness of apoptosis in combating baculovirus infection. The results of these experiments indicate that apoptosis can be an extremely powerful response to baculovirus infection, reducing viral replication, infectivity, and the ability of the virus to spread within the insect host even if a successful infection is established. Apoptosis is especially effective when it is combined with other innate antiviral defenses, which are largely unexplored in insects to date. PMID- 15791954 TI - The role of host cell death in Salmonella infections. AB - Salmonella enterica is an important enteric pathogen of humans and a variety of domestic and wild animals. Infection is initiated in the intestinal tract, and severe disease produces widespread destruction of the intestinal mucosa. Salmonella strains can also disseminate from the intestine and produce serious, sometimes fatal infections with considerable cytopathology in a number of systemic organs. A combination of bacterial genetic and cell biology studies have shown that Salmonella uses specific virulence mechanisms to induce host cell death during infection. Salmonella produces one set of virulence proteins to promote invasion of the intestine and a different set to mediate systemic disease. Significantly, each set of virulence factors mediates a distinct mechanism of host cell death. The Salmonella pathogenicity island-1 (SPI-1) locus encodes a type III protein secretion system (TTSS) that delivers effector proteins required for intestinal invasion and the production of enteritis. The SPI-1 effector SipB activates caspase-1 in macrophages, releasing IL-1beta and IL 18 and inducing rapid cell death by a mechanism that has features of both apoptosis and necrosis. Caspase-1 is required for Salmonella to infect Peyer's patches and disseminate to systemic tissues in mice. Progressive Salmonella infection in mice requires the SPI-2 TTSS and associated effector proteins as well as the SpvB cytotoxin. Apoptosis of macrophages in the liver is found during systemic infection. In cell culture, Salmonella strains induce delayed apoptosis dependent on SPI-2 function in macrophages from a variety of sources. This delayed apoptosis also requires activation of TLR4 on macrophages by the bacterial LPS. Downstream activation of kinase pathways leads to balanced pro- and antiapoptotic regulatory factors in the cell. NF-kappaB and p38 mitogen activated protein kinase (MAPK) are particularly important for the induction of antiapoptotic factors, whereas the kinase PKR is required for bacterial-induced apoptosis. The Salmonella SPI-2 TTSS is essential for altering the balance in favor of apoptosis during intracellular infection, but the effectors involved remain poorly characterized. The SpvB cytotoxin has been shown to play a role in apoptosis in human macrophages by depolymerizing the actin cytoskeleton. A model for the role of bacteria-induced host cell death in Salmonella pathogenesis is proposed. In the intestine, the Salmonella SPI-1 TTSS and SipB mediate macrophage death by caspase-1 activation, which also releases IL-1beta and IL-18, promoting inflammation and subsequent phagocytosis by incoming macrophages and leading to dissemination to systemic tissues. Intracellular secretion of virulence effector proteins by the SPI-2 TTSS facilitates growth of Salmonella in these macrophages and the delayed onset of apoptosis in extraintestinal tissues. These infected, apoptotic cells are targeted for engulfment by incoming macrophages, thus perpetuating the cycle of cell-to-cell spread that is the hallmark of systemic Salmonella infection. PMID- 15791955 TI - Role of macrophage apoptosis in the pathogenesis of Yersinia. AB - Yersinia species that are pathogenic for humans (Yersinia pestis, Yersinia pseudotuberculosis, and Yersinia enterocolitica) induce apoptosis in macrophages. Yersinia-induced apoptosis utilizes the mitochondrial pathway and is executed by activation of caspase cascades. The mechanism of Yersinia-induced apoptosis in macrophages has two essential components. One component is the innate immune response of macrophages to the pathogen, which leads to the activation of a survival response and a death response. Recognition of the bacterial cell envelope component lipopolysaccharide by Toll-like receptor 4 (TLR4) constitutes an important part of the innate immune response to the pathogen. The second essential component is YopJ, a protein secreted into Yersinia-infected macrophages via a bacterial type III secretion system, which selectively shuts down the survival pathway. In the absence of the survival pathway, the death pathway is executed, and Yersinia-infected macrophages undergo apoptosis. In this review, we introduce the basic features of Yersinia pathogenesis, summarize our current understanding of Yersinia-induced apoptosis, and discuss the role of apoptosis during Yersinia infection. PMID- 15791956 TI - Entamoeba histolytica activates host cell caspases during contact-dependent cell killing. AB - Entamoeba histolytica is a human intestinal parasite that causes amoebic colitis as well as liver abscesses. Host tissues are damaged through a three-step process involving adherence, contact-dependent cytolysis, and phagocytosis. These three processes all contribute to the pathogenicity of this parasite. Adherence is provided by the Gal/GalNAc adherence lectin. Host cells are lysed in a contact dependent fashion. There is evidence that suggests that this contact-dependent killing involves the induction of the host cell's apoptotic machinery. Phagocytosis can then occur, consistent with metazoan apoptotic clearance. PMID- 15791957 TI - Interactions between malaria and mosquitoes: the role of apoptosis in parasite establishment and vector response to infection. AB - Malaria parasites of the genus Plasmodium are transmitted from host to host by mosquitoes. Sexual reproduction occurs in the blood meal and the resultant motile zygote, the ookinete, migrates through the midgut epithelium and transforms to an oocyst under the basal lamina. After sporogony, sporozoites are released into the mosquito haemocoel and invade the salivary gland before injection when next the mosquito feeds on a host. Interactions between parasite and vector occur at all stages of the establishment and development of the parasite and some of these result in the death of parasite and host cells by apoptosis. Infection-induced programmed cell death occurs in patches of follicular epithelial cells in the ovary, resulting in follicle resorption and thus a reduction in egg production. We argue that fecundity reduction will result in a change in resource partitioning that may benefit the parasite. Apoptosis also occurs in cells of the midgut epithelium that have been invaded by the parasite and are subsequently expelled into the midgut. In addition, the parasite itself dies by a process of programmed cell death (PCD) in the lumen of the midgut before invasion has occurred. Caspase-like activity has been detected in the cytoplasm of the ookinetes, despite the absence of genes homologous to caspases in the genome of this, or any, unicellular eukaryote. The putative involvement of other cysteine proteases in ancient apoptotic pathways is discussed. Potential signal pathways for induction of apoptosis in the host and parasite are reviewed and we consider the evidence that nitric oxide may play a role in this induction. Finally, we consider the hypothesis that death of some parasites in the midgut will limit infection and thus prevent vector death before the parasites have developed into mature sporozoites. PMID- 15791958 TI - Apoptosis and its modulation during infection with Toxoplasma gondii: molecular mechanisms and role in pathogenesis. AB - Infection with the obligate intracellular protozoan Toxoplasma gondii leads to lifelong persistence of the parasite in its mammalian hosts including humans. Apoptosis plays crucial roles in the interaction between the host and the parasite. This includes innate and adaptive defense mechanisms to restrict intracellular parasite replication as well as regulatory functions to modulate the host's immune response. Not surprisingly, however, T. gondii also extensively modifies apoptosis of its own host cell or of uninfected bystander cells. After infection, apoptosis is triggered in T lymphocytes and other leukocytes, thereby leading to suppressed immune responses to the parasite. T cell apoptosis may be largely mediated by Fas engagement but also occurs independently of Fas under certain conditions. Depending on the magnitude of T cell apoptosis, it is either associated with unrestricted parasite replication and severe pathology or facilitates a stable parasite-host-interaction. However, T. gondii has also evolved strategies to inhibit host cell apoptosis. Apoptosis is blocked by indirect mechanisms in uninfected bystander cells, thereby modulating the inflammatory response to the parasite. In contrast, inhibition of apoptosis in infected host cells by direct interference with apoptosis-signaling cascades is thought to facilitate the intracellular development of T. gondii. Blockade of apoptosis by intracellular parasites may be achieved by different means including interference with the caspase cascade, increased expression of antiapoptotic molecules by infected host cells, and a decreased activity of the poly(ADP ribose) polymerase. The intriguing dual activity of T. gondii to both promote and inhibit apoptosis requires a tight regulation to promote a stable parasite host interaction and establishment of persistent toxoplasmosis. PMID- 15791959 TI - Modulation of the immune response in the nervous system by rabies virus. AB - Rabies virus (RABV) is a pathogen well-adapted to the nervous system, where it infects neurons. RABV is transmitted by the bite of an infected animal. It enters the nervous system via a motor neuron through the neuromuscular junction, or via a sensory nerve through nerve spindles. It then travels from one neuron to the next, along the spinal cord to the brain and the salivary glands. The virions are then excreted in the saliva of the animal and can be transmitted to another host by bite. Thus preservation of neuronal network integrity is crucial for the virus to be transmitted. Successful invasion of the nervous system by RABV seems to be the result of a subversive strategy based on the survival of infected neurons. This strategy includes protection against virus-mediated apoptosis and destruction of T cells that invade the CNS in response to infection. PMID- 15791960 TI - Apoptotic cells at the crossroads of tolerance and immunity. AB - Clearance of apoptotic cells by phagocytes can result in either anti-inflammatory and immunosuppressive effects or prostimulatory consequences through presentation of cell-associated antigens to T cells. The differences in outcome are due to the conditions under which apoptosis is induced, the type of phagocytic cell, the nature of the receptors involved in apoptotic cell capture, and the milieu in which phagocytosis of apoptotic cells takes place. Preferential ligation of specific receptors on professional antigen-presenting cells (dendritic cells) has been proposed to induce potentially tolerogenic signals. On the other hand, dendritic cells can efficiently process and present antigens from pathogen infected apoptotic cells to T cells. In this review, we discuss how apoptotic cells manipulate immunity through interactions with dendritic cells. PMID- 15791961 TI - Maintenance of certification and competency. PMID- 15791962 TI - Robotic systems and surgical education. AB - This experimental study aimed at evaluating the efficiency of robots in the learning of surgical techniques. We recruited 40 surgeons, divided them into 2 groups of 20, each of which used the robotic system. The first group consisted of experienced physicians, and the second group comprised physicians in training. Each surgeon was allowed to use the da Vinci robotic system for 30 minutes twice in the span of 24 hours. The practice time period was divided into 15 minutes for tying and placement of sutures and 15 minutes for incisions and vascular suturing. We recorded the times required for the performances, and a statistically significant outcome was obtained. With variance analysis (ANOVA), it has been shown that the time needed to perform the exercises depends in a statistically significant way on the kind of test to be performed (P<0.01), the experience of the surgeon (P<0.001), and the kind of operation (P<0.025). Robotic systems can be an optimal tool both for residents and experienced surgeons, for learning of basic surgical tasks and for perfection of clinical skills. The use of the system has great potential in surgical training, offering a reduction in the learning period, enabling checking for errors, and allowing an evaluation of the capabilities obtained. Final goals are a drastic reduction in the learning curve, a better technique, with a significant reduction in surgical errors and complications, with greater safety for the patient. PMID- 15791963 TI - Total laparoscopic hysterectomy utilizing a robotic surgical system. AB - OBJECTIVES: To describe the use of a robotic surgical system for total laparoscopic hysterectomy. METHODS: We report a series of laparoscopic hysterectomies performed using the da Vinci Robotic Surgical System. Participants were women eligible for hysterectomy by standard laparoscopy. Operative times and complications are reported. RESULTS: We completed 10 total laparoscopic hysterectomies between November 2001 and December 2002 with the use of the da Vinci Robotic Surgical System. Operative results were similar to those of standard laparoscopic hysterectomy. Operative time varied from 2 hours 28 minutes to 4 hours 37 minutes. Blood loss varied from 25 mL to 350 mL. Uterine weights varied from 49 g to 227 g. A cystotomy occurred in a patient with a history of a prior cystotomy unrelated to the robotic system. CONCLUSION: Total laparoscopic hysterectomy is a complex surgical procedure requiring advanced laparoscopic skills. Tasks like lysis of adhesions, suturing, and knot tying were enhanced with the robotic surgical system, thus providing unique advantages over existing standard laparoscopy. Total laparoscopic hysterectomy can be performed using robotic surgical systems. PMID- 15791964 TI - Fertility considerations in laparoscopic treatment of infiltrative bowel endometriosis. AB - OBJECTIVE: The purpose of this study was to examine our experience with laparoscopic and laparoscopically assisted management of bowel endometriosis and to recommend treatment approaches, considering patient goals for both pain mitigation or fertility, or both. METHODS: The medical records of 187 women treated laparoscopically for intestinal endometriosis were reviewed retrospectively for presenting symptoms, methods of surgical treatment, complications, and efficacy of treating pain and infertility. The extent of resection was determined by the severity of the endometriotic lesion, tempered by the patient's fertility goals. RESULTS: The most common patient complaint preceding surgery was pelvic pain. In addition, 58 (31%) patients experienced impaired fertility. Of the patients available for long-term follow-up, 152 (85%) reported complete or significant long-term pain relief. Complete pain relief in the immediate postoperative period was significantly more likely with partial bowel resection compared with shaving only, 92% vs 80%, respectively, P<0.04. The least invasive procedure, shaving, was associated with a significantly lower complication rate, 6%, compared with 23% for disc excision (P<0.007) and 38% for segmental resection (P<0.001), and higher pregnancy rates. The incidence of pregnancy in patients with a history of infertility was 34% during the follow-up period. PMID- 15791966 TI - Randomized comparison between two microlaparoscopic techniques for partial salpingectomy. AB - OBJECTIVE: We compared 2 techniques for performing a partial salpingectomy by using microlaparoscopy and either bipolar coagulation or loop ligation. METHODS: A 3-mm transumbilical laparoscope with secondary midline port sites midway and suprapubically was used to perform a partial salpingectomy in 109 women desiring permanent sterilization. Each patient was randomly assigned to undergo a tubal resection either after Pomeroy ligation (n= 54) or after bipolar coagulation with Kleppinger forceps (n=55). Postoperative pain, as assessed using a 10-point visual analog scale, was the primary comparison endpoint. RESULTS: No technical difficulties with either technique required conversion to a minilaparotomy. The mean time to remove both tubal segments was not different between techniques (7 minutes, 21 seconds; range, 4 minutes, 25 seconds to 15 minutes, 43 seconds). Each segment (mean, 1.6 cm; range, 0.8 to 3.5 cm) was confirmed in the operating room, then histologically. Postoperative pain at 6 hours was scored similarly (median, ligation 4.6, coagulation 4.0 of 10). Outpatient recovery was the same, unless pelvic pain required overnight observation (ligation, 4 patients; coagulation, 2 patients). CONCLUSION: Partial salpingectomy, using microlaparoscopy with either bipolar coagulation or loop ligation, was performed with comparable ease, confirmation of the removed tube, and similar postoperative discomfort. PMID- 15791965 TI - Interstitial cystitis and endometriosis in patients with chronic pelvic pain: The "Evil Twins" syndrome. AB - OBJECTIVE: To determine the prevalence of interstitial cystitis and endometriosis in patients with chronic pelvic pain. METHODS: A prospective analysis was conducted in 178 women with CPP who presented with bladder base/anterior vaginal wall and/or uterine tenderness, with or without irritative voiding symptoms. The Potassium Sensitivity Test was used to assess bladder epithelial dysfunction. Patients were evaluated with concurrent laparoscopy and cystoscopy with hydrodistention. RESULTS: Laparoscopic findings among the 178 patients with chronic pelvic pain supported a diagnosis of endometriosis in 134 (75%) patients, and cystoscopy confirmed a diagnosis of interstitial cystitis in 159 (89%) patients. Both interstitial cystitis and endometriosis were diagnosed in 115 patients (65%). The Potassium Sensitivity Test was positive in 146 (82%) patients, with 140 (96%) of these patients diagnosed with interstitial cystitis and 105 (72%) with endometriosis. CONCLUSIONS: Results of this prospective study show that interstitial cystitis and endometriosis may frequently coexist in patients with chronic pelvic pain. A positive Potassium Sensitivity Test accurately predicted the presence of interstitial cystitis in 96% of these patients with chronic pelvic pain, as confirmed by cystoscopic hydrodistention. It is necessary to consider the diagnosis of endometriosis and interstitial cystitis concurrently in the evaluation of patients with chronic pelvic pain to avoid unnecessary delay in identifying either condition. PMID- 15791967 TI - Laparoscopic nonanatomic hepatic resection employing the LigaSure device. AB - BACKGROUND: Advancements in technology have allowed laparoscopic surgery to expand into advanced procedures such as liver resection; however, the value and safety of laparoscopic liver surgery is still a topic for debate. This study was designed to evaluate the feasibility and outcome of laparoscopic nonanatomic hepatectomy using the LigaSure device in a swine model. METHODS: Nonanatomic hepatic lobe resection was performed in 3 groups comparing the open finger fracture method, the open method with the LigaSure device, and the laparoscopic method with the LigaSure device. The cut surfaces of the liver were evaluated for bleeding and biliary leakage at the time of the operation. The animals were inspected 48 hours later for hemorrhage and evaluated with cholescintigraphy (hepatobiliary iminodiacetic acid [HIDA] scan) for biliary leakage, in addition to histological evaluation of liver specimens. RESULTS: No hemorrhage or biliary leakage was noted in the groups where the LigaSure device was used, whereas 1 animal from the open finger-fracture method sustained a bile leak observed on HIDA scan. The operative blood loss was considerably less in the groups where the LigaSure device was used, and the shortest operative time was observed in the laparoscopic group. CONCLUSIONS: The LigaSure device can be safely and effectively used to perform a laparoscopic nonanatomic hepatectomy. PMID- 15791968 TI - An innovative bipolar instrument for laparoscopic surgery. AB - OBJECTIVE: Bipolar electrosurgery is an excellent method for obtaining hemostasis at laparoscopy. The present study describes and evaluates a bipolar device that can be more versatile and cost effective in advanced operative procedures than the traditional instrumentation. METHODS: This was a retrospective, case controlled analysis of bipolar instrumentation with a design classification of II 2. A single surgeon in a private practice setting performed all procedures. Sixteen patients, matched for age and pathology were evaluated by videotape review to determine the comparative efficiency of the BiCOAG bipolar dissector/grasper versus traditional Kleppinger bipolar forceps. Efficiency here is defined as comparative operating times in each group. RESULTS: The number of instrument changes per case was counted because this appeared to be the only variable other than time that differentiated the 2 groups. The BiCOAG bipolar dissector/grasper device group had 4 times fewer instrument changes and significantly decreased operating room times when compared with that of the Kleppinger forceps group. Because cost per unit of operating room time was a constant figure, the decrease in cost that resulted due to the decrease in operating time is considered a cost-effective measure. CONCLUSIONS: The BiCOAG bipolar dissector/grasper is a cost-effective, efficient instrument for use in operative laparoscopic procedures. PMID- 15791970 TI - Laparoscopic incisional hernia repair in obese patients. AB - BACKGROUND AND OBJECTIVES: Laparoscopic incisional hernia repair is coming to the forefront as a preferred method of repair due to the advantages offered by minimally invasive techniques. To evaluate safety and feasibility of this approach in obese patients when performed by a general surgeon trained in basic laparoscopy with no prior experience in this technique, we reviewed our early experience in the first 18 patients. METHODS: All patients with incisional hernias presenting to a single surgeon from 2000 to 2002 were offered laparoscopic repair. Patients were informed about the limited experience of the surgeon in this particular field. Those who consented were repaired laparoscopically using a standard 4-port technique, one 12-mm port and three 5-mm ports. All patients with body mass index > or =30 were included in this review. A retrospective review of the data included demographics, operative time, blood loss, hospital stay, postoperative complications, and patient satisfaction. RESULTS: Nineteen laparoscopic repairs were completed in 18 patients. No conversions to open repair were necessary. All patients were females except for 2. All hernia sacs were left in place, some of which were empty while others required extensive lysis of adhesions to release sac contents. Mean fascial defect was 102.5 cm2. One defect was closed primarily without mesh, while the rest were closed using Composix mesh in 1 and Dual Plus Gore-Tex mesh in the rest. Three patients were discharged from the recovery room. Mean follow-up was 24 months. No wound or mesh infections occurred. Eight patients had no complications. Eight patients had asymptomatic seromas. Two patients had hematomas; none of them required drainage. One patient had nonspecific dizziness. One patient presented with bowel obstruction secondary to early recurrence (within a week). The repair was salvaged laparoscopically. Upon evaluation by telephone calls, all patients indicated extreme satisfaction with the results. CONCLUSIONS: A general surgeon with training in basic laparoscopy can safely perform laparoscopic incisional hernia repair on obese patients with minimal complications. The procedure requires a short leaning curve of no more than 3 cases and few extra materials to be feasible at any hospital in the US. Patient satisfaction with this technique is certainly gratifying. PMID- 15791969 TI - Endoscope-assisted inguinal hernia repair. AB - BACKGROUND: Since the advent of laparoscopic inguinal hernia repair, the procedure has invited numerous controversies, and although the procedure has some definitive advantages, no definitive indications for its use have been formulated. The objective of this study was to investigate a novel method for inguinal hernia repair (through a small 2 cm to 2.5 cm) single skin incision that combines the time-tested fundamentals of Lichtenstein's tension-free repair with the advantages of laparoscopic assistance. METHODS: The study was conducted as a randomized, controlled trial over a 1-year period and included 50 patients. Only patients with simple reducible hernias without associated comorbid conditions were included. The patients were randomized into 2 groups of 25 patients each. One group underwent conventional tension-free meshplasty, while the other group underwent the repair through a single 2-cm to 2.5-cm skin incision with laparoscopic assistance. This repair was carried out with the help of an indigenously designed steel retractor, 10-mm laparoscope, and conventional instruments; the mesh was fixed with the help of endotacks. Univariate analysis of variance techniques using SPSS 7.5 software was used for data analysis. RESULTS: Two groups were compared for time taken for the procedure, size of skin incision, postoperative pain, complications, return to work, and cosmetic appearance. The results showed a significant decrease in postoperative pain and an earlier return to work, along with much improved cosmesis for the new procedure. CONCLUSIONS: Although the study was conducted with a limited number of patients and a very short follow-up, it is worth considering this method over laparoscopic and conventional techniques, especially in reducible hernias. PMID- 15791971 TI - Laparoscopic "Dome-down" cholecystectomy with the LCS-5 Harmonic scalpel. AB - OBJECTIVE: Misidentification of ductal anatomy and electrocautery injuries are complications associated with laparoscopic cholecystectomy (LC). Dome-down LC creates a 360-degree view of the gallbladder-cystic duct junction, reducing the risk for anatomy misidentification. In addition, ultrasonic instrumentation eliminates the risk for electrocautery injuries. This study assessed the feasibility and safety of dome-down LC combined with ultrasound technology. METHODS: Patients with noncancerous gallbladder disease were enrolled consecutively. Gallbladders were classified by clarity (Class I to IV) of anatomy and pathology (acute, chronic, or acalculous). The gallbladder was dissected from the gallbladder bed using a dome-down technique, and the cystic artery was coagulated and transected with the LCS-5 Harmonic scalpel (Ethicon Endo-Surgery Inc., Cincinnati, Ohio). The cystic duct was ligated with 2-polydioxanone Endoloops size 2-0 and sharply divided, leaving one Endoloop on the cystic duct stump. RESULTS: LC was successfully completed in 105 patients (mean age, 44 years; range, 18 to 91 years) in whom the anatomy was classified as Class I in 30 (29%) patients, Class II in 42 (38%), Class III in 25 (24%), and Class IV in 8 (8%). Gallbladder dissection time ranged from 8 to 42 minutes (mean, 18 min). The operating room time ranged from 32 to 128 minutes (mean, 55 min). Two gallbladder perforations occurred, but no complications were associated with the extrahepatic biliary tree, viscera, or major blood vessels. Elective conversion occurred in 8 (7.6%) patients due to poor visualization of anatomy because of inflammation and adhesions. Patient blood loss was minimal in all cases. No postoperative complications were observed after a 6-month follow-up. CONCLUSION: Dome-down laparoscopic cholecystectomy with the LCS-5 Harmonic scalpel decreases the potential for misidentification of ductal anatomy, has minimal complications, and eliminates electrocautery risks. Conversion is related to poor visualization of anatomy due to inflammation and adhesions. PMID- 15791972 TI - Laparoscopic colectomy in the management of large, sessile, transformed colorectal polyps. AB - BACKGROUND: Colonic polyps are usually removed endoscopically. Surgical intervention is reserved for large, inaccessible colorectal polyps that have underdone malignant transformation. Laparoscopic management of colonic polyps has gained a well-defined role. METHODS: Since 1993, 650 laparoscopic colectomies have been performed in our department. Twenty-eight patients with large, sessile, polyps that have undergone malignant transformation underwent elective laparoscopic colectomy. Operative procedures included 14 sigmoidectomies, 10 low anterior recto-sigmoid resections, 3 right colectomies, and 1 left colectomy. Central ligation of vessels and lymph node dissection were preformed in all patients. RESULTS: The main outcome measures include conversion rate (11%), morbidity (11%), and mortality (3.5%). The mean return of bowel function was 3.1 days, liquid intake 1.4 days, solid food intake 2.5 days, and mean hospital stay 8 days. The mean specimen length was 23 cm, and the mean number of retrieved lymph nodes was 15. Malignancy according to Dukes classification was in situ, n=4; A, n=15; B, n=4; C, n=4; and D, n=1. During follow-up, 2 patients developed liver metastases. CONCLUSION: Laparoscopic colectomy is a technically feasible and effective method for treating large colorectal polyps that have undergone malignant transformation. PMID- 15791973 TI - Factors associated with complications of open versus laparoscopic sigmoid resection for diverticulitis. AB - BACKGROUND: This study critically reviews sigmoid colon resection for diverticulitis comparing open and laparoscopic techniques. METHODS: We conducted a retrospective review of all open and laparoscopic cases of diverticulitis between 1992 and 2001. Data analyzed included the following: indications for operation, postoperative complications, and incidence of laparoscopic conversion to laparotomy. Major and minor complications were analyzed in relation to patients' preoperative diagnosis, age, presence or absence of splenic flexure mobilization, length of stay, and laparoscopic sigmoid resection versus open sigmoid resection. RESULTS: Over a 10-year period, 166 resections for diverticulitis were performed including 126 open cases and 40 laparoscopic cases. No significant differences existed in patient characteristics between the groups. Major complications occurred in 14% of patients, and the laparoscopic conversion rate was 20%. The presence of abscess, fistula, or stricture preoperatively was associated with a higher complication rate only in patients > or =50 years old undergoing open sigmoid resection. The length of stay between patients undergoing laparoscopic resection was significantly less than in patients having open resection. CONCLUSION: Advanced laparoscopic sigmoid resection is an alternative to open sigmoid resection in patients with diverticulitis and its complications. Open sigmoid resection in patients >50 years may have a higher complication rate in complicated diverticulitis when compared with laparoscopic sigmoid resection (all patient ages) and open sigmoid resection (patients <50 years old). Regarding complications, no difference existed between the length of stay in patients with open vs. laparoscopic resection. PMID- 15791974 TI - Indications for the laparoscopic treatment of gastroesophageal reflux disease. AB - BACKGROUND: The aim of the study was to evaluate the indications and surgical techniques for the treatment of gastroesophageal reflux disease. METHODS: From 1998 through 2000, we performed gastroesophageal reflux surgery on 12 patients. Preoperative studies revealed third-degree esophagitis in most patients with no functional alterations of the esophagus itself. The patients underwent a laparoscopic Nissen-Rossetti fundoplication. No conversions to open laparotomy were necessary. The mean operative time was 180 minutes. RESULTS: No intraoperative or immediate postoperative complications occurred. The mean postoperative stay was 6.2 days. Transitory postoperative dysphagia was noted in 8 patients. In 5 patients, it was mild and regressed after 2 weeks; in 3 cases, it was severe and regressed over 2 months. CONCLUSION: A laparoscopic surgical approach is a satisfactory method for correcting gastroesophageal reflux disease. The efficacy of medical therapy has been well established. However, in the young person who may be required to take medication for many years or in those persons who are intolerant of standard medical therapy for gastroesophageal reflux disease, a surgical intervention is a satisfactory option. The success of medical therapy can be used as a predictive criterion of the success of laparoscopic Nissen-Rosetti fundoplication when normal motility of the esophageal corpus is present. PMID- 15791975 TI - A new technique for laparoscopic anterior resection for rectal endometriosis. AB - BACKGROUND: Anterior rectal resection is sometimes necessary to treat deeply infiltrating rectovaginal endometriosis. We describe a completely laparoscopic approach as a new way of excising rectal endometriosis that can be used without opening any part of the rectum. This avoids opening the abdomen or any risk of fecal spillage. METHODS: The patient received preoperative oral bowel preparation. Ureteric stents (6 F) were inserted cystoscopically. The peritoneum in the ovarian fossae was opened lateral to any disease and the rectum reflected off the back of the cervix, leaving any endometriosis on the front of the rectum. The pelvic peritoneum was reflected medially, below the level of the ureters. The mesorectum was then dissected off a 6-cm length of rectum by using a Harmonic scalpel. A circular end-to-end anastomosis instrument was passed anally until the outline of the anvil was visible, inside the colon, above the diseased rectum. The anvil was detached and held by a soft grasper before the rectum was then divided above and below the disease using a laparoscopic stapling device. The tip of the anvil was pushed through the proximal end of the colon allowing reanastomosis of the rectal stump. CONCLUSION: The patient was discharged after 5 days without complications. PMID- 15791976 TI - Synchronous rectovaginal, urinary bladder, and pulmonary endometriosis. AB - BACKGROUND: Extragenital endometriosis is an uncommon condition that can affect almost any organ system and tissue in the human body. Disease involving multiple distant sites is extremely uncommon. METHODS: We report a rare case of synchronous rectovaginal, urinary bladder, and pulmonary endometriosis. We performed a Medline literature search using keywords "endom etriosis," "recto vaginal," "pulmonary," "bladder," "ureteral," "bowel," "extrapelvic," and "extragenital" and were unable to find any prior case reports of such findings. A 31-year-old female presented with catamenial dysuria of 1-year duration, pleurisy associated with spontaneous pneumothoraces of 7 months' duration and a long standing history of pelvic pain. A multispecialty team with experience in endoscopic techniques was assembled, consisting of a thoracic, a urologic, and a gynecologic surgeon. Video-assisted thoracoscopic surgery with fulguration of all visible pleural endometriosis and pleurodesis was performed, followed by laparoscopic segmental bladder wall endometrioma excision and resection of rectovaginal endometriosis. Twelve months after surgery and without additional hormonal treatment, the patient is symptom free. CONCLUSION: Extragenital endometriosis may coexist in multiple sites. A high index of suspicion aids in the diagnosis. A multidisciplinary approach in a tertiary center, followed by appropriate surgical eradication of visible disease, can successfully treat endometriosis even in such extreme cases. PMID- 15791977 TI - Robot-assisted laparoscopic partial nephrectomy. AB - The indications for nephron-sparing surgery and for minimally invasive surgery are continually expanding. Nephron-sparing surgery, also known as partial nephrectomy, presents a challenge to the minimally invasive surgeon. Herein, we describe our technique of robot-assisted laparoscopic partial nephrectomy. This approach may have potential advantages of including easier excision and suturing. Moderate training is required. PMID- 15791978 TI - Portal vein thrombosis: an unusual complication of laparoscopic cholecystectomy. AB - BACKGROUND: Complications following laparoscopic cholecystectomy are encountered infrequently due to increasing proficiency in laparoscopic surgery. The occurrence of portal venous thrombosis following laparoscopic cholecystectomy has not been previously described and forms the basis of this report. METHODS: A healthy, 32-year-old, female on oral contraceptives underwent an uneventful laparoscopic cholecystectomy for symptomatic gallbladder disease. Sequential compression devices and mini-dose unfractionated heparin were used before the procedure. The patient was discharged home on the first postoperative day without complaints. She returned 1 week later with nausea, bloating, and diffuse abdominal pain. RESULTS: Ultrasonography of the abdomen revealed thrombosis of the portal vein not seen in the preoperative ultrasound and the superior mesenteric vein. Computer tomography of the abdomen and pelvis on the same day confirmed this finding and showed a wedge-shaped infarction of the right lobe of the liver. The patient was anticoagulated with intravenous heparin. An extensive coagulation workup revealed elevation of the Immunoglobulin G anticardiolipin antibody. A percutaneous transhepatic portal vein thrombectomy was performed. A postprocedure duplex ultrasound of the abdomen demonstrated recannalization of the portal venous system with no flow voids. Anticoagulation therapy was continued, and the patient was discharged home with resolution of her ileus. She was maintained on a therapeutic dose of warfarin. CONCLUSIONS: This case demonstrates an unusual complication of laparoscopic cholecystectomy. It may have resulted from the use of oral contraceptives, elevation of the Immunoglobulin G anticardiolipin antibody, unrecognized trauma, and was accentuated by the pneumoperitoneum generated for the performance of the laparoscopic cholecystectomy. Our case report provides insight and poses questions regarding necessary perioperative measures for thromboprophylaxis in young females on oral contraceptives undergoing elective laparoscopic abdominal surgery. PMID- 15791979 TI - Gastric perforation secondary to ingestion of a sharp foreign body. AB - Foreign body ingestions are commonly seen by pediatricians and emergency room personnel. The majority of foreign bodies transit through the gastrointestinal system without any complications. Perforations are uncommon and usually involve peritonitis. We present a case of gastric perforation secondary to ingestion of a sharp metallic pin. Our patient did not demonstrate any of the classic findings described in the literature. His complaints, physical examination, and radiological findings all pointed to acalculous cholecystitis or biliary dyskinesia as the source of pain. Laparoscopy, however, provided the definitive diagnosis. Gastroduodenal perforation secondary to foreign body ingestion should be included in the differential diagnosis of these atypical cases of abdominal pain, especially if such history can be elicited from the patient. PMID- 15791980 TI - Laparoscopic ventral hernia repair using a two (5-mm) port technique. AB - OBJECTIVE: High recurrence rates have been documented after primary repair of incisional hernias. Laparoscopic ventral and incisional hernia repairs have been performed with very low rates of recurrence. We have modified the standard technique of laparoscopic repair in patients with small incisional and ventral hernias. The purpose of this study was to document the technique utilizing only two 5-mm ports and demonstrate that it is safe, effective, and feasible. METHODS: Three patients with small incisional or ventral hernias were examined. The standard laparoscopic ventral hernia repair technique was modified as follows: two 5-mm ports were inserted on opposite sides of the defect. The defects ranged from 2.5 cm to 4 cm in size. Expanded polytetrafluoroethylene mesh (DualMesh, WL Gore, Flagstaff, AZ) was used to cover the hernia defect, overlapping the defect margins circumferentially by 3 cm. The mesh diameter ranged from 8.5 cm to 10 cm. The mesh was inserted through a 5-mm skin incision site and affixed into position with transfascial sutures and spiral tacks. RESULTS: The operative time ranged from 53 minutes to 57 minutes. All patients were discharged home the day of surgery and reported minimal postoperative pain. Follow-up ranged from 6 months to 1 year; all patients were doing well without recurrence. CONCLUSION: Laparoscopic repair of ventral or incisional hernias can be performed using only two 5-mm ports. This technique can be done on an outpatient basis in a safe, timely fashion. PMID- 15791981 TI - Retroperitoneal pyelolithotomy for management of renal calculi. AB - OBJECTIVE: We evaluated the role of retroperitoneoscopic pyelolithotomy in the management of renal calculi. METHODS: Fifty-six cases (male, 27; female, 29) of solitary or multiple renal calculi were evaluated in the study. There were 46 patients with a single calculus, 4 patients with a staghorn calculus, and 6 with a caliceal calculus. Retroperitoneoscopic pyelolithotomy was carried out after creating a retroperitoneal space with the balloon dissection method. Pneumoretroperitoneum was maintained by carbon dioxide insufflation. RESULTS: Stone clearance was achieved in all cases barring 2 cases of caliceal calculi that were converted to the open procedure. The postoperative hospital stay averaged 4 days. Patients were ambulatory within 24 hours and back to work within 7 days on average. Complications encountered were peritoneal rent, subcutaneous emphysema, and superficial wound infection. The postoperative analgesic requirement averaged 100 mg of diclofenac (2 tablets). CONCLUSIONS: Retroperitoneoscopic pyelolithotomy is a safe, simple, and effective minimally invasive procedure and is a feasible option that can be recommended for management of renal calculi. PMID- 15791982 TI - Early lap-band erosion associated with colonic inflammation: a case report and literature review. AB - INTRODUCTION: Laparoscopic adjustable gastric banding is an effective and safe surgical modality for the treatment of morbid obesity. Erosion of the band into the stomach has been reported. No reports are available on erosion of the Lap Band following diverticulitis of the colon. CASE REPORT: A 31-year-old female with a body mass index (BMI) of 52 underwent an uneventful laparoscopic Lap-Band placement. Postoperative contrast study revealed good positioning of the band and no evidence of leakage. The patient's recovery was uneventful except for an elevated temperature of 101.5 degrees F that was attributed to her atelectasis. She had lost 52 lbs. and remained asymptomatic for 3 months. Following this period of successful weight loss, she presented with complaints of abdominal pain for 3 days associated with diarrhea of 7 days' duration. A Gastrografin contrast study showed no evidence of a leak or band slippage but erosion was suspected. Upper endoscopy confirmed erosion of the band into the stomach. Computed tomography (CT) of the abdomen revealed thickening of the sigmoid and descending colon with mesenteric fat stranding consistent with diverticulitis. Laparoscopic removal of the Lap-Band system was performed. CONCLUSION: We postulate that colonic diverticulitis could have been a precipitating factor in the development of band erosion. Intraabdominal sepsis resulting in subacute infection of the Lab Band system may be the underlying factor. PMID- 15791984 TI - Analysis and physics of laparoscopic intracorporeal square-knot tying. AB - Square knots are often used in open surgery to approximate tissue borders or tie off tubular structures like vessels or ducts. Three common methods are used for surgical square-knot tying: one-hand tying, two-hand tying, and the instrument tying technique. Two types of suture placements are studied in both the open and laparoscopic surgical fields. The first called equal length has suture segment ends placed at equal distances from the tying site. The second called unequal length has one suture end further away from the tying site than the other. Laparoscopic intracorporeal square-knot tying maneuvers are analyzed herein. Mechanical analysis of square-knot tying movements reveals that regardless of location or method used in construction, all square knots consist of 2 half knots. For study purposes, these sets of movements are identified in laparoscopy as maneuver A and maneuver B. Further breakout of these maneuvers reveals that they consist of 5 motions. This study reveals that 16 different ways exist to place a square knot by means of the laparoscopic intracorporeal technique. It is likely that difficulty mastering this essential skill is not just the result of poor instrumentation, improper port placement, or the limitations of a 2 dimensional video image. It may also be attributed to mixing up the different square-knot tying techniques during random practice exercises. This is possible if the surgeon is ignorant of the technical variations present in what most people consider a simple task. PMID- 15791983 TI - Laparoscopic multifunctional instruments: design and testing of initial prototypes. AB - BACKGROUND: Advances in minimally invasive surgical techniques will require new types of instrument end-effectors for smaller, longer, and flexible instruments. These include a new class of multifunctional instruments capable of performing more than 1 task with a single set of working jaws. Furthermore, it is desired that multifunctional instruments be designed to provide improved dexterity compared with that in currently commercially available instruments. METHODS: Three prototypes of multifunctional laparoscopic surgical instruments are described: (1) a mechanical scissors-grasper, (2) a mechanical scissors grasperarticulator, and (3) a compliant mechanism scissors-grasper. Methods of baseline analysis, design methods and considerations, and subjective evaluations of interim prototypes are presented. RESULTS: The 3 prototypes demonstrate promising early results. However, based on subjective evaluation, these prototypes do not perform individual functions as well as basic disposable single function laparoscopic instruments do. CONCLUSIONS: The concept of multifunctionality and increased end-effector dexterity is achievable as demonstrated by the prototypes presented. Further work is required to refine, simplify, and improve the multifunctional instruments to a point where they may be useful as surgical tools. PMID- 15791985 TI - In situ determination of refractive index and size of Bacillus spores by light transmission. AB - Light-extinction measurements in the wavelength range of 400 to 1000 nm are performed in situ on Bacillus subtilis endospores during heat-shock-induced activation. Simultaneous information on particle size and refractive indices during activation is calculated from the transmission spectra by use of the Gaussian ray approximation of anomalous diffraction theory. During activation the refractive index of the core decreases from 1.51 to 1.39, and the size increases from 0.38 to 0.6 microm. PMID- 15791986 TI - Switchable holographic gratings formed in polymer-dispersed liquid-crystal cells by use of a He-Ne laser. AB - We report on the formation of switchable holographic transmission gratings in polymer-dispersed liquid-crystal cells by use of the 633-nm wavelength of a He-Ne laser. We present results for the structure, diffraction efficiency, and switching characteristics of the gratings. PMID- 15791987 TI - Radiating and nonradiating dispersion-managed solitons. AB - Using spectral analysis, we present the salient features of the radiating behavior of optical solitons in dispersion-managed fiber systems. Depending on the map strength of the system, we find a rich variety of dynamic states that includes weak, moderate, and strong radiation states. We establish the existence of a critical map strength at which a Gaussian-shaped light pulse with a large initial dressing undergoes highly stable nonradiative propagation over transoceanic distances. PMID- 15791988 TI - Nanostructured optical fiber with surface-enhanced Raman scattering functionality. AB - A powerful method for the production of reproducible surface-enhanced Raman scattering (SERS) substrates is described based on the scaling properties of glass rods when drawn into fibers. The fabrication process involves chemically eroding the cleaved tips of drawn silica imaging fibers and then coating them with silver. For an appropriate choice of final diameter the drawn and eroded tips show clearly defined and regular triangular formations on a scale of approximately 80 nm. The favorable SERS properties of these structures have been demonstrated by the observation of enhancement factors of approximately 10(6). PMID- 15791989 TI - Two-loop-based low-coherence multiplexing fiber-optic sensor network with a Michelson optical path demodulator. AB - A two-loop-based low-coherence multiplexing fiber-optic sensor network is proposed and demonstrated. It greatly extends the multiplexing capacity of this kind of sensor. A practical implementation of this scheme uses a popular amplified spontaneous emission light source and standard single-mode fiber, which are commonly used in the communications industry. The sensor's two-loop topology is completely passive. Absolute length measurements can be obtained for each sensing fiber segment for use in measuring the quasi-distribution strain or temperature. For large-scale smart structures this technique not only extends the multiplexing potential but also provides a redundancy for the sensing system. This means that the two-loop sensor network can break down at several points and the sensing system will still work even if some of the embedded two-loop sensors have been destroyed. The robustness of the nine-sensor two-loop sensing network is investigated and discussed. PMID- 15791990 TI - Parallel demodulation system and signal-processing method for extrinsic Fabry Perot interferometer and fiber Bragg grating sensors. AB - A parallel demodulation system for extrinsic Fabry-Perot interferometer (EFPI) and fiber Bragg grating (FBG) sensors is presented that is based on a Michelson interferometer and combines the methods of low-coherence interference and Fourier transform spectrum. Signals from EFPI and FBG sensors are obtained simultaneously by scanning one arm of a Michelson interferometer, and an algorithm model is established to process the signals and retrieve both the wavelength of the FBG and the cavity length of the EFPI at the same time, which are then used to determine the strain and temperature. PMID- 15791991 TI - High-temperature-resistant chemical composition Bragg gratings in Er3+-doped optical fiber. AB - Chemical composition gratings (CCGs), unlike standard fiber Bragg gratings (FBGs), do not suffer a significant decrease in reflectance or an irreversible wavelength shift when they are exposed to elevated temperatures. To date, the growth of CCGs has been related to the fluorine content of the fibers in which they are written. It is shown that FBGs with high thermal stability, resembling CCGs, can be fabricated in Er3+-doped optical fibers that do not contain any fluorine. PMID- 15791992 TI - Holographic recording with reduced intermodulation noise in periodically poled lithium niobate. AB - We show that the use of periodically poled lithium niobate doped with Fe and Y ensures a considerable improvement in the quality of reconstructed images compared with the use of single-domain crystals. This improvement is due to inhibition of intermodulation noise and elimination of optical damage. PMID- 15791993 TI - Three-dimensional display with a long viewing distance by use of integral photography. AB - We developed a technique of three-dimensional (3-D) display for distant viewing of a 3-D image without the need for special glasses. The photobased integral photography (IP) method allows precise 3-D images to be displayed at long viewing distances without any influence from deviated or distorted lenses in a lens array. We calculate elemental images from a referential viewing area for each lens and project the corresponding result images to photographic film through each lens. We succeed in creating an image display that appears to have three dimensionality even when viewed from a distance, with an image depth of 5.7 m or more in front of the display and 3.5 m or more behind the display. To the best of our knowledge, the long-distance IP display presented here is technically unique because it is the first report of generation of an image with such a long viewing distance. PMID- 15791994 TI - Marangoni forces created by surface plasmon decay. AB - We present optical microfluidic manipulation of silicone oil and glycerol via surface tension driven forces sustained by surface plasmon deexcitation energy. The phonon energy associated with the decaying optically excited surface plasmons in a thin gold foil creates thermal gradients capable of actuating fluid flows. Spectral dependence of the plasmon decay length and control of optical beam characteristics are shown to provide a means for further manipulation. PMID- 15791995 TI - Multiwavelength fiber ring lasing by use of a semiconductor optical amplifier. AB - A multiwavelength fiber ring laser obtained by use of a semiconductor optical amplifier (SOA) with a simple laser cavity configuration is reported. A Fabry Perot filter was used in the fiber laser ring cavity to achieve more than 50 simultaneous wavelength lasing oscillations with a frequency separation of 50 GHz. The resulting stable broadband multiwavelength lasing operation was attributed to broadband and flat gain of the SOA, which has a gain flatness of 0.8 dB for more than 20 nm. The laser has a total output power of -3 dBm and a signal-to-spontaneous-noise ratio of 30 dB. PMID- 15791996 TI - Monitoring the thermally induced structural transitions of collagen by use of second-harmonic generation microscopy. AB - The thermal disruption of collagen I in rat tail tendon is investigated with second-harmonic generation (SHG) microscopy. We investigate its effects on SHG images and intensity in the temperature range 25 degrees-60 degrees C. We find that the SHG signal decreases rapidly starting at 45 degrees C. However, SHG imaging reveals that breakage of collagen fibers is not evident until 57 degrees C and worsens with increasing temperature. At 57 degrees C, structures of both molten and fibrous collagen exist, and the disruption of collagen appears to be complete at 60 degrees C. Our results suggest that, in addition to intensity measurement, SHG imaging is necessary for monitoring details of thermally induced changes in collagen structures in biomedical applications. PMID- 15791997 TI - In vivo dark-field reflection-mode photoacoustic microscopy. AB - Reflection-mode photoacoustic microscopy with dark-field laser pulse illumination and high-numerical-aperture ultrasonic detection is designed and implemented in noninvasively imaged blood vessels in the skin in vivo. Dark-field optical illumination minimizes the interference caused by strong photoacoustic signals from superficial structures. A high-numerical-aperture acoustic lens provides high lateral resolution, 45-120 microm in this system. A broadband ultrasonic detection system provides high axial resolution, estimated to be approximately 15 microm. The optical illumination and ultrasonic detection are in a coaxial confocal configuration for optimal image quality. The system is capable of imaging optical-absorption contrast as deep as 3 mm in biological tissue. PMID- 15791998 TI - Photothermal image flow cytometry in vivo. AB - The capability of photothermal (PT) microscopy to image moving, unlabeled cells in real time in vivo is demonstrated in a study of circulating red and white blood cells in blood and lymph microvessels of rat mesentery. Potential applications of this optical tool, called PT flow cytometry, are discussed. PMID- 15791999 TI - High diffraction efficiency and a quasi-permanent grating in photorefractive nematic liquid crystal at low temperature. AB - At a temperature lower than the melting point of pure pentyl-cyanobiphenyl liquid crystal (5CB) we studied the photorefractive effect of 5CB heavily doped with fullerene (C60) and obtained a diffraction efficiency of the photorefractive grating of more than 40%, which is more than twice as high as the best data reported to our knowledge. A quasi-permanent grating was observed with a diffraction efficiency of 11.2% after the grating was kept in the dark for a month. PMID- 15792000 TI - Tandem chirped quasi-phase-matching grating optical parametric amplifier design for simultaneous group delay and gain control. AB - We present a broadband optical parametric amplifier design using tapered gain and tandem chirped quasi-phase-matching gratings to obtain flat gain and group-delay spectra suitable for applications such as ultrashort-pulse amplification and fiber-optic communication systems. Although a tapered-gain amplifier consisting of a single chirped grating can provide constant gain over a wide frequency range, it cannot be used to control the group delay across the spectrum. We propose controlling both the gain and the group delay profiles using a two-stage amplifier configuration, in which the idler of the first is used as the input signal of the second. PMID- 15792001 TI - Soliton spiraling in optically induced rotating Bessel lattices. AB - We address soliton spiraling in optical lattices induced by multiple coherent Bessel beams and show that the dynamic nature of such lattices makes it possible for them to drag different soliton structures, setting them into rotation. We can control the rotation rate by varying the topological charges of lattice-inducing Bessel beams. PMID- 15792002 TI - Terahertz generation with tandem seeded optical parametric generators. AB - A simple difference frequency generation (DFG) scheme based on two seeded optical parametric generators is presented as a tunable terahertz (THz) source. Using the nonlinear optical crystal 4-dimethylamino-N-methyl-4-stilbazolium-tosylate (DAST) as the DFG crystal, our system has demonstrated continuous and seamless tunable operation from 1.6 to 4.5 THz. The output bandwidth of the THz source is 2.4 GHz. The utility of the source over this spectral range is demonstrated by measuring a high-resolution transmission spectrum of water vapor in air. PMID- 15792003 TI - Performance of nonlinear photonic crystal devices at high bit rates. AB - I investigate theoretically the performance limits for all-optical switching devices incorporating photonic crystals made of nonlinear materials operating at high bit rates. I compare two switching techniques--shifting of the photonic bandgap and the more traditional interferometric method enhanced by slow wave propagation in photonic crystals-and show that the latter always has an advantage. I also demonstrate that the benefits provided by the photonic crystal in the interferometer are still severely limited by the dispersion and become significant only in materials combining high nonlinearity, high index contrast, and high damage threshold. PMID- 15792004 TI - Iodine stabilization of a diode laser in the optical communication band. AB - The iodine molecule has frequently been used as a frequency reference from the green to the near-infrared wavelength region (500-900 nm). We describe the frequency locking of the second-harmonic signal of a 197.2-THz (1520.25-nm) distributed-feedback diode laser to the absorption lines of the iodine hyperfine structure; a frequency jitter below 0.1 MHz was achieved at a 300-ms time constant. This scheme provides a simple, compact, and high-performance frequency reference in the optical communication band. PMID- 15792005 TI - Simplex optimization method for illumination design. AB - A modified simplex optimization method is developed for the design of illumination systems. The simplex method is a judicious choice for illumination optimization because of its robustness and convergence properties. To optimize the simplex method, its four parameters are adjusted dependent on the dimensionality of the space to converge with fewer iterations. This work is presented for the end game, when the optimizer is converging on a local optimum rather than searching for it. Up to a 37% reduction in the number of computations is realized. An example using a compound parabolic concentrator is compared between the standard and the modified simplex methods, providing over 22% improvement in the end game. PMID- 15792006 TI - Silicon microlens structures fabricated by scanning-probe gray-scale oxidation. AB - We report on the micromachining of silicon microlens structures by use of scanning-probe gray-scale anodic oxidation along with dry anisotropic etching. Convex, concave, and arbitrarily shaped silicon microlenses with diameters as small as 2 microm are demonstrated. We also confirm the high fidelity of pattern transfer between the probe-induced oxides and the etched silicon microlens structures. Besides the flexibility, the important features of scanning-probe gray-scale anodic oxidation are small pixel size and pitch (of the order of tens of nanometers), an unlimited number of gray-scale levels, and the possibility of creating arbitrarily designed microlens structures with exquisite precision and resolution. With this approach, refractive, diffractive, and hybrid microlens arrays can be developed to create innovative optical components. PMID- 15792007 TI - Two-photon fourth-order polarimetery. AB - We demonstrate a fourth-order polarimeter that employs two-photon detection to measure the fourth-order polarization coherency matrix (FOCM) of an optical signal. We measure the FOCM of picosecond pulses subjected to polarization mode dispersion (PMD). The fourth-order polarimeter distinguishes different PMD states even when the state of polarization, including the degree of polarization, is constant. PMID- 15792008 TI - Generalization of the rainbow Airy theory to nonuniform spheres. AB - Rainbow techniques permit measurement of refractive indices, and hence the temperatures of liquid droplets through determination of the absolute angular position of a rainbow interference image in space. The Airy theory, which is commonly used to explain the rainbow effect, permits the determination of a unique refractive-index value, even in the presence of nonuniformities in the droplet. An extension of this theory to spheres that exhibit internal refractive index gradients is proposed. The case of burning droplets is considered as an example of such spheres, and the results obtained are successfully compared with those presented in the literature. PMID- 15792009 TI - Radiation produced by femtosecond laser-plasma interaction during dielectric breakdown. AB - Optical breakdown by femtosecond and nanosecond laser pulses in transparent dielectrics produces an ionized region of dense plasma confined within the bulk of the material. This ionized region is responsible for broadband radiation that accompanies the breakdown process. Spectroscopic measurements of the accompanying light have been used to show that, depending on the laser parameters, the spectra may originate from plasma-induced second-harmonic generation, supercontinuum generation, or thermal emission by the plasma. By monitoring the emission from the ionized region, one can ascertain the predominant breakdown mechanism and the morphology of the damage region. PMID- 15792010 TI - Disentangling the volume effect through intensity-difference spectra: application to laser-induced dissociation of H2+. AB - An intensity-difference spectrum method is developed to disentangle the intensity volume effect inherent in focused laser beam interaction with gas-phase matter. This method is applicable to a Gaussian beam of constant axial intensity, which keeps the exact contribution from a predetermined intensity range and eliminates the contributions from lower intensities. We apply this method to the angularly resolved kinetic energy release spectrum of laser-induced dissociation of H2+. The difference spectrum at higher intensities is found to be dominated by the bond-softening process, and the distribution shifts to lower energy and becomes narrower with increasing intensity. PMID- 15792011 TI - Femtosecond-laser-based synthesis of ultrastable microwave signals from optical frequency references. AB - We use femtosecond laser frequency combs to convert optical frequency references to the microwave domain, where we demonstrate the synthesis of 10-GHz signals having a fractional frequency instability of < or =3.5 x 10(-15) at a 1-s averaging time, limited by the optical reference. The residual instability and phase noise of the femtosecond-laser-based frequency synthesizers are 6.5 x 10( 16) at 1 s and -98 dBc/Hz at a 1-Hz offset from the 10-GHz carrier, respectively. The timing jitter of the microwave signals is 3.3 fs. PMID- 15792012 TI - 40-gHz, 100-fs stimulated-Brillouin-scattering-free pulse generation by combining a mode-locked laser diode and a dispersion-decreasing fiber. AB - A 40-GHz, 100-fs pulse train was successfully generated by soliton compression of a mode-locked laser diode (MLLD) pulse with a dispersion-decreasing fiber. The MLLD had a longitudinal mode linewidth as broad as 60 MHz, which made it possible to suppress stimulated Brillouin scattering and achieve stable, ultrahigh-speed pulse compression without applying external frequency modulation. PMID- 15792013 TI - Effect of spherical aberration and surface waves on propagation of lens-coupled terahertz pulses. AB - Spatiotemporal measurements of a near-single-cycle terahertz pulse emitted from a photoconductive switch terahertz (THz) source show the effects of spherical aberration and surface waves on the pulse shape. The measured phase front has a swallow-tail shape described by catastrophe theory that contributes to the concentric ring structure of THz beam profiles. A time-of-flight model shows that the pulse shape is due to propagation along a cusp caustic and enhancement of the wings of the swallow-tail pulse is caused by surface waves. PMID- 15792014 TI - [The effect of vitamin, macro and microelement supplementation on markers of iron status in blood of pregnant women]. AB - Interest in assessing iron status in the population of women during their child bearing age has grown recently both in terms of detecting iron stores and evaluating whether they are sufficient to meet increasing needs during pregnancy. The fact that iron deficiency anaemia often develops in pregnancy indicates that that iron stores and dietary iron are insufficient to requirements and iron supplementation is needed. The aim of the present study was to determine the effect of vitamin, macro and microelement supplementation on iron status of pregnant women and of their newborns. It is shown that serum total iron concentration in the group of women without supplementation decreased in the III trimester to 60% of that observed after 12 weeks of pregnancy (p < 0.0001). In supplemented group the decrease was not statistically significant. Increased value of transferin in III trimester in comparison to the I trimester by 38% (p < 0.0001) and by 55% (p < 0.0001) respectively in non supplemented and supplemented groups were found. It was accompanied by significant increase of total iron binding capacity (TIBC) (p < 0.0001). In both studied groups the level of ferritin was lower in the III than in the I trimester. In serum of women without supplementation, ferritin concentration decreased by 35% (p < 0.003); whereas in supplemented group it was statistically insignificant. In serum of umbilical cord blood concentration of total iron, transferin and ferritin were similar in newborns of both groups of mothers. The above results suggested that vitamin, macro and microelement supplementation of pregnant women improved iron status in mothers and in their children. It seems that administration Vibovitmama formula can be recommended as a dietary supplement in pregnancy in order to prevent subclinical deficiency of iron. PMID- 15792015 TI - [The effect of vitamin-mineral supplementation on vitamins D, A (beta-carotene) and E concentration in blood of matched maternal-cord pairs]. AB - The aim of the present study was to investigate the effect of Vibovitmama supplementation on status of fat soluble vitamins D, A (beta-carotene) and E in blood of pregnant women and in umbilical cord blood of their children. In group taking Vibovitmama serum concentration of vitamin D, was higher by 10% than in the placebo group was observed. However it was within the normal range and was not statistically significant. Supplementation affected vitamin D concentration in serum of cord blood. It was higher by 20% in the group taking vitamin preparation than in group taking placebo and was statistically significant (p < 0.05). Retinol and beta-carotene in plasma increased during pregnancy by 15% and 25% (p < 0.002) respectively in placebo and supplemented group. In umbilical cord blood in the group taking Vibovitmama, concentration of vitamin A was 1.52 micromol/L and of beta-carotene 1.86 micromol/L versus 1.45 micromol/L and 1.79 micro/L found in group of placebo. In plasma level, vitamin E was higher in late pregnancy than in the I trimester (by about 40%) and was not modified by vitamins supplementation. In erythrocytes the concentration of tocopherol increased by 18% in placebo and by 25% in group taking Vibovitmama. In the plasma of cord blood level of vitamin E was not modified by supplementation, whereas in red blood cells it was slightly increased (by 10%) but not statistically significant. Our results suggest that vitamin-mineral supplementation of pregnant women improved status of fat soluble vitamins in mothers and their children. PMID- 15792016 TI - [The effect of vitamin-mineral supplementation on the level of MDA and activity of glutathione peroxidase and superoxide dismutase in blood of matched maternal cord pairs]. AB - The aim of the present study was to investigate the effect of VIBOVITmama (Polfa Kutno S.A.) supplementation on lipid peroxidation (MDA) and activity of superoxide dismutase (SOD) and selenium-dependent glutathione peroxidase (GPx) in blood of pregnant women and in umbilical cord blood of their children. It is shown, that MDA concentration in plasma increased during pregnancy in both studied groups; however in vitamin-mineral C supplemented women it was lower (p < 0.05) than in placebo (p < 0.005). In erythrocytes MDA level was within the same range in both groups, but in women taking vibovit it was (in late pregnancy) lower by 10% than in trimester I. Activity of GPx in red blood cells decreased during pregnancy. In placebo the fall was two-fold more intensive (16%) than in the supplemented group (p < 0.005). Activity of erythrocytes SOD slightly increased (5%) in placebo, whereas in the group taking the formula it decreased (4%) but not statistically significant. MDA level in plasma of cord blood was not modified by supplementation, but in red blood cells was lowered by 20% (97.3 nmol/gHb-supplemented vs 116.3 nmol/gHb-placebo) which was observed. In erythrocytes of umbilical cord blood activity of GPx was higher in newborns of women taking vibovit (31.9 U/gHb) than in the control group (29.7 U/gHb), whereas activity of SOD was similar. Our result suggested that vitamin-mineral supplementation of pregnant women improved antioxidant status in mothers and their children. It seems that in order to prevent oxidative damage VIBOVITmama formula can be recommended as a supplement diet in pregnant women. PMID- 15792017 TI - [Effect of vitamin-mineral supplementation on the status of some microelements in pregnant women]. AB - We studied the effect of VIBOVITmama (Polfa Kutno S.A.) supplementation on the course of pregnancy duration, delivery and puerperium and also the status of zinc, copper and selenium in the blood of matched maternal-cord pairs. Healthy pregnant women (n=138) were divided by a double blinded trial into a test group taking vitamin and mineral supplementation containing 15 mg of zinc, 2 mg of copper and 20 microg of selenium and a control group taking placebo. Course of pregnancy, delivery and puerperium were analyzed, as well as concentration of bioelements in the blood serum of pregnant women in the I, II, and II trimester and in umbilical cord blood of their children. In the study group we noted fewer by 1.45% cases of pregnancy induced hypertension and a much higher rate of natural deliveries (75%) in comparison to the control group (53%). During pregnancy in the supplemented group a 1% increase of zinc serum concentration was found. In the control group zinc concentration decreased by 7% (p < 0.005). Insignificant differences in serum concentration of copper and selenium was observed between the study group and controls. In umbilical cord blood the differences in the concentration of the above microelements were also insignificant. The results allow for the statement that VIBOVITmama supplementation stabilizes the zinc, copper and selenium levels in blood of pregnant women without the risk of overdosing, especially with reference to selenium. PMID- 15792018 TI - [Dietary assessment of pregnant women--vitamin and mineral supplementation- justified or not?]. AB - The aim of the study was to assess the dietary intake of energy and nutrients of pregnant women in the Outpatient Clinic of Obstetrics and Gynecology of National Research Institute of Mother and Child in Warsaw. The nutrient intake, the average daily food rations and their nutritional value were evaluated on the basis of recall and record of 7-days menus. The average daily food intake was compared to the recommendations for the first and second half of pregnancy. It was found, that 44% of women changed dietary habits during pregnancy. The nutritional value of their daily food intake was closer to recommendations in the first half of pregnancy than in the second half. The food intake of pregnant women with supplementation (S-group) and taking placebo (NS-group) were also compared. In the first half of pregnancy the average food intake of women from S group and NS-group was similar. In the second half of pregnancy the nutrient intake of S-group women was closer to recommendations than in the second group. However the deficiency of certain B vitamins and microelements (calcium, iron, zinc, magnesium) occurred in both groups. The intake of vitamin A, sodium and phosphorus exceeded the recommendations. The results of the study support the need of supplementation in the pregnant women diets with microelements and vitamins. PMID- 15792019 TI - [Intake of pharmacologic supplements of vitamins and minerals during pregnancy. Survey conducted in Krakow]. AB - The aim of this study was to describe pharmacological supplements during pregnancy in the sample of 406 nonsmoking women, residents of Krakow. Multivitamins' supplements were taken by 79.1%, folic acid by 41.4%, magnesium by 23.9% and iron only by 14.5% of the study group. Our results confirm that vitamin and mineral supplements, ferrum, folic acid and magnesium in particular, are inadequate in comparison to recommended intakes. Women with lower education (secondary school or lower) failed to supplement diet with multivitamins (OR = 5.74; 95% CI: 1.41-23.5) and folic acid (OR = 2.11; 95% CI: 1.22-3.66). Otherwise, health problems during the previous pregnancies and nulliparity have lowered pharmacological folic acid supplements (OR = 0.45; 95% CI: 0.21-0.97 and OR = 0.57; 95% CI: 0.32-0.99, respectively). Results of our study show that diet assessed for pregnancy period does not meet nutritional requirements. Pharmacological supplementary intake of vitamins and minerals is also insufficient. It is advisable to popularize education concerning necessity and usefulness of vitamin and microelement supplementation during medical examinations in the childbearing age. PMID- 15792020 TI - [Auditory brainstem responses to logon and tone burst 1000 Hz frequency stimuli in infants--own experience]. AB - The use of frequency-specific stimuli in recording auditory brainstem responses (ABR) allows for hearing objectivity concerning the given frequency. Non-linear intensity growth (logon) and linear intensity growth (tone burst), 1000 Hz frequency short tone stimuli are most commonly used. Differences between hearing threshold values and other ABR measures in recordings obtained after stimulation with broad-band click and logon were analysed in a group of high-risk infants. 71 infants aged 1 to 9 months (36 girls and 35 boys) were included in the group. Broadband envelope shaped 2000-4000 Hz repeated 5 times per second clicks, acquisition and analyse time 12.5 ms and Gauss shaped 1000 Hz logon and tone bursts repeated 4 times per second, acquisition and analyse time 20.0 ms were used as stimuli. Click induced mean threshold values were the same after stimulation with logon, tone bursts and clicks. Wave 1, III and V latencies decreased with stimulus frequency. Mean amplitudes of waves I, III and V obtained after logon stimulation were slightly higher than after tone burst stimulation. PMID- 15792021 TI - [End-tidal sevoflurane concentrations for laryngeal mask airway insertion and tracheal intubation in children]. AB - BACKGROUND: Sevoflurane, a new inhalation anesthetic agent characterized by a low blood-gas partition coefficient and nonpungent odor, appears suitable as an induction agent for children. This study was conducted to determine the sevoflurane concentration required for tracheal intubation and insertion laryngeal mask in children. METHODS: Thirty patients, aged 3-7 years, scheduled for surgery during general anesthesia were randomly assigned into two groups: Group I (n=15)--tracheal intubation (IT), Group II (n=15)--laryngeal mask insertion (LMI). After the predetermined end-tidal concentration of the sevoflurane, tracheal intubation or laryngeal mask insertion was attempted without neuromuscular relaxants. RESULTS: Sevoflurane MAC-LMI was 1.53 +/- 0.23%, sevoflurane MAC-TI was 2.35 +/- 0.31%, significantly greater than MAC-LMI. CONCLUSIONS: Laryngeal mask insertion can be performed at a lesser sevoflurane concentration than that required for tracheal intubation. PMID- 15792022 TI - [Concentration of cadmium in breast cancer tissue of women living in the Wielkopoiska region]. AB - The research was carried out concerning the contents of cadmium in breast cancer tissues in 52 women aged 35-81 years (x = 67.62 +/- 12.43) living in the Wielkopolska region. Particular concentrations of this metal in the range of 0.01 1.08 mg/g wet wt. (x = 0.13 +/- 0.18) were found in all the studied specimens. The obtained results were assessed regarding the women's place of residence- Kalisz, Leszno, Pila or Poznan voivodships--and a particular living area: an industrialised town, other towns or villages of the Wielkopolska region. A statistically significant difference in the content of cadmium in breast cancer tissues was found in women inhabiting the Poznan and Pila voivodships. Moreover, marked differences were observed between the average cadmium concentrations in the breast cancer tissue samples in women living in the remaining voivodships and the types of living areas. PMID- 15792023 TI - [The influence of L-arginine on secretion of nitric oxide and haemodynamic function of isolated rat's heart subjected to cold cardioplegic ischemia and reperfusion]. AB - BACKGROUND: L-arginine is a precursor of nitric oxide (NO), which is formed endogenously as a result of reaction catalyzed by synthase NO (NOS). Administration of L-arginine to cardioplegic solution St. Thomas Hospital N 2 or used during reperfusion of heart can facilitate the return of left ventricle function after hypothermic ischaemia through increase of release of nitric oxide and preservation of endothelial cells functions. Positive effects of these researches could have extremely beneficial application in clinical practice, improving results of cardiosurgical operations conducted in extracorporeal circulation. OBJECTIVE: How addition of L-arginine to cardioplegic solution or during reperfusion influences secretion of nitric oxide, coronary flow and haemodynamic function of heart. METHODS: The research was conducted on isolated heart model of rats (Wistar inbreeding). The research was carried out in order: 20-minutes initial perfusion, 30-minutes cold cardioplegic arrest with or without addition of L-arginine and reperfusion with Krebs-Henseleit's solution with or without addition of L-arginine. RESULTS: Addition of L-arginine in the preischemic period and during reperfusion stimulates increase of nitric oxide secretion, what positively correlates with increasing of coronary flow. Addition of L-arginine to cardioplegic solution significantly increases coronary flow and cardiac output what improves postischaemic haemodynamic function of heart. PMID- 15792024 TI - [Informed consent for angiography and endovascular treatment]. AB - The aim of this paper was to evaluate the methods of passing information concerning angiography and endovascular treatment from doctors to patients. Patients undergoing modern procedures are especially prone to anxiety or even panic caused by the lack of sufficient information about the planned procedure. There are many reasons for such phenomena. One of them is extremely fast improvement of new, sophisticated technology and lack of humane attitude to patients. Authors examined 60 patients before and after angiography and endovascular treatment. Most of them (92%) knew nothing about the procedures. 30% of the patients gained information from other patients. After angiography and endovascular treatment most of patients assessed the procedures as less dangerous than expected. Our questionnaire is attached to this paper. PMID- 15792025 TI - [Imaging of dialysis fistulae]. AB - Vascular access malfunctions can be diagnosed by means of imaging and non-imaging methods. The former are: ultrasound (US), colour Doppler ultrasonography, arteriography and phlebography. Radiological imaging allows for direct assessment of the morphology and function of the hemodialysis fistula, and shows precisely the kind and location of lesions, that may lead to access failure. The aim of the study was to present the algorithm of methods employed in evaluation of hemodialysis fistulae, that had been developed by the authors. From March 1995 to July 2001, 1020 colour Doppler ultrasonography examinations were performed, followed by 250 angiographies of hemodialysis fistulae. In the studied group there were 170 well functioning hemo-dialysis fistulae. They were evaluated in order to establish normal blood flow parameters in the fistula. In 135 patients with symptoms of fistula failure US revealed the presence of pathological lesions, such as thrombosis or aneurysm of the venous branch. In 193 cases the presence of stenosis was stated. Three hundred eleven hemodialysis fistulae were sonographically evaluated before the patient was referred to surgical (81) or intravascular (230) treatment. Post-operative assessment was performed in 125 cases. In 231 cases, angiography confirmed the results of US. In 19 cases the degree of fistula stenosis was underestimated in colour Doppler sonography. Colour Doppler sonography is a method of choice in evaluation of arteriovenous hemodialysis fistulae. Abnormal results of physical examinations, dialysis parameters, as well as patients complaints associated with the fistula are indications to perform an ultrasound examination. Angiography is required in case of dubious US findings or as the first step in endovascular treatment. PMID- 15792026 TI - [Flow cytometric analysis of type 1 and type 2 subsets of CD4+ and CD8+ cells in intermittent allergic rhinitis and chronic urticaria--methodological aspects]. AB - Human immune cells, included T helper cells and T cytotoxic/suppressor cells, may be divided into two distinct subtypes according to the profile of secreted cytokines. The allergic disorders are characterised by type 2 lymphocyte response, while the autoimmune diseases are associated with type 1 response. The aim of this study was to assess the usefulness of flow cytometric analysis of intracellular expression of cytokines typical for type 2 (IL-4) and type 1 response (IFN-gamma) in helper and cytotoxic/suppressor T cells in patients with chronic urticaria or intermittent allergic rhinitis. 10 subjects (6 male) with intermittent allergic rhinitis (a group ALER) and 6 subjects (1 male) with chronic urticaria (a group PP) were included into the study. The intracellular expression of IL-4 and IFN-gamma in CD4+ or CD8+ cells after stimulation with ionomycin/PMA was estimated by flow cytometer (FACSCalibur, Becton Dickinson) and serum levels of both cytokines were assessed with ELISA (R & D Systems) in all subjects. The percentage of CD4+ cell producing IFN-gamma was statistically higher in the PP group than in the ALER group, and the percentage of IL-4 producing CD8+ cells was also higher, although non significantly. The intracellular expression of IL-4 was comparable in both cell populations in the two examined groups. The surface expression of CD4 diminished after stimulation with PMA in all subjects. No correlations between serum level and intracellular expression of the examined cytokines were observed. CONCLUSION: A tendency to the predominance of type 1 response of T lymphocytes was observed in chronic urticaria compared to intermittent allergic rhinitis. PMID- 15792027 TI - [Risk assessment of patients and preparation for abdominal aortic aneurysm repair -the effect of collaboration among the internist, anaesthesiologist and surgeon]. AB - Abdominal aortic aneurysm (AAA) is a common cardiovascular disease of increasing incidence. The complex clinical assessment with regard to indications for surgical treatment is necessary for each patient with AAA diagnosis. Taking into consideration the high postoperative complication risk and mortality rate among these patients, the general evaluation especially the function of cardiovascular system together with surgical and anaesthesiological management are required. Proper cooperation with specialists of internal medicine and often with cardiologist is essential for reduction of perioperative risk as a consequence of treatment, that improves the function of the most important organs, in the best possible manner. The engagement of specialists in internal medicine, anaesthesiology and surgery enables the patient to carry through the operation in safe conditions. We describe perioperative risk factors and basic principles of patient evaluation and preparation for AAA reconstructive surgery. PMID- 15792028 TI - [Mucous membrane involvement in the course of autoimmune blistering skin diseases and drug eruptions. Differential diagnosis]. AB - In the following paper the most common drug eruptions and auto-immune, blistering skin diseases in which mucous lesions precede skin lesions development are discussed. Non-characteristic clinical pictures of mucous lesions result in diagnostic difficulties of these diseases, and prognosis is dependent on prompt and precise diagnosis. The aim of the work was to describe clinical features of mucous membranes lesions and point to differential diagnosis. PMID- 15792029 TI - [Patent foramen ovale--demographic, clinical and echocardiographic characteristics]. AB - Patent foramen ovale (PFO) represents the most common persistent abnormality of fetal origin. The aim of the study was to analyze the incidence and characteristics of PFO and to assess PFO as the risk factor of paradoxical embolism. 945 consecutive pts (442F and 503M aged 13-85, mean 53.02 +/- 14.42 years) in whom TEE was performed and flow through PFO was detected in basal conditions (colour-Doppler) and/or during provocation (Valsalva/cough manoeuvre) with contrast (saline infusion) at the period 1998-2002 was retrospectively analyzed. 183 pt had a history of systemic embolisation (SE) or transient ischemic attack (TIA). PFO was diagnosed [PFO(+)] in 57 pts (6%)-22F, 35M aged 14 77 years, mean 52.44 +/- 14.84 years. 888 pts without PFO [PFO(-)] created a control group. Two subgroups were analyzed in the study group: xPFO(+) - 28 pts with PFO as the only abnormality, yPFO(+) - 29 pts with PFO and other heart/aortic disease. Similarly, in the control group two subgroups were analysed: xPFO(-) - pts with normal TEE and negative history of heart/aortic disease and yPFO(-) - pts without PFO but with other heart/aortic disease. Aneurysmal formation of interatrial septum was detected in 24 pts, and in 6 pts it coexisted with PFO. The intensity of shunt was significantly higher in yPFO(+) than in xPFO(+) group. In 5 pts permanent shunt through PFO at the basal stage was observed (4 pts with significant regurgitation of the atrio-ventricular valves and one patient with tricuspid stenosis). Percentage of pts with the history of SE/TIA was significantly higher in group xPFO(+) than in xPFO(-) (p < 0.01). CONCLUSIONS: (1) PFO is a risk factor of SE/TIA only in group xPFO(+). (2) Flow through PFO in group xPFO(+) is generally minimal. (3) In case of coexisting heart pathology PFO may provoke significant right-to-left or even left-to-right shunt. PMID- 15792031 TI - Perhaps just a bad dream. PMID- 15792030 TI - [Laryngeal mask airway after failed intubation. Case report]. AB - The case of a 58-year old patient, who nearly experienced cardiac arrest due to growing oxygen deficiency, was presented. The arrest was brought about by the impossibility to perform endotracheal intubation. A laryngeal mask successfully replaced an intubation tube and enabled artificial lung ventilation and restoration of correct blood oxygenation. PMID- 15792032 TI - Body handlers after terrorism in Oklahoma City: predictors of posttraumatic stress and other symptoms. AB - Posttraumatic stress and depressive symptoms were assessed in 51 body handlers after Oklahoma City's 1995 terrorist bombing. Although many handlers were inexperienced and knew someone killed, symptoms were low postdisaster and decreased significantly after 1 year. Higher symptomatology and seeking mental health treatment correlated with increases in alcohol use and new physical problems but not with demographics, exposure, or experience. Four respondents with the highest posttraumatic stress symptoms at both time points reported high physical and alcohol use problems and mental health treatment use, suggesting that these should be carefully assessed in body handlers postdisaster. Coping techniques are described, as well as possible reasons for unexpected resilience in the majority. PMID- 15792033 TI - Exposure, threat appraisal, and lost confidence as predictors of PTSD symptoms following September 11, 2001. AB - Six months after September 11, 2001 (9/11), 124 New York City workers participated in a self-report study of symptoms of posttraumatic stress disorder (PTSD). Although direct exposure to the terrorist attacks of 9/11 was limited, estimates of the prevalence of current PTSD in this mostly ethnic minority population ranged from 7.8% to 21.2%, as measured by the PTSD Checklist (F. W. Weathers, B. T. Litz, D. S. Herman, J. A. Huska, & T. M. Keane, 1993). Consistent with the study hypotheses, direct exposure to the attacks of 9/11, worries about future terrorist attacks (threat appraisal), and reduced confidence in self after 9/11 each predicted symptoms of PTSD, even after controlling for symptoms of anxiety and depression. These results support the idea that a traumatic event's meaning is associated with PTSD symptoms. Gender was not a significant predictor of symptoms, once other demographic variables were controlled. Most respondents who met the criteria for current PTSD had not sought therapy or counseling. PMID- 15792034 TI - Preparing for bioterrorism at the state level: report of an informal survey. AB - Members of 18 states' departments of mental health were interviewed about their plans for managing the psychosocial impacts of a bioterrorism event. Questions were developed from recommendations of an international conference on planning for bioterrorism ("Planning for bioterrorism," 2000). Information derived from the survey highlights the need for, and the importance of, mental health consultation to the state's planning process. Familiarity with the unique psychological and behavioral consequences of a bioterrorism event in contrast to natural disasters is essential. Realistic training scenarios that incorporate likely psychosocial impacts and appropriate mental health response must be developed. PMID- 15792035 TI - The long-term mental health effects of nuclear trauma in recent Russian immigrants in the United States. AB - The long-term mental health sequelae (15 years) of the 1986 Chernobyl nuclear disaster in the former Soviet Union (FSU) were assessed in survivors migrated to the United States, as part of an ongoing study. Using a retrospective, cross sectional design with standardized Russian mental health measures, a sample of 261 Russians residing in the New York tristate area were tested for multiple domains of mental health functioning. Multivariate analyses pointed to aspects of the accident and its negative repercussions as reliable predictors of current psychological distress. Russians who had lived closer to the disaster, and had greater exposure to it, currently experience higher levels of anxiety (p < .0004) and posttraumatic reactions (p < .03) than those who had lived at a further distance. These findings are consistent with reports of long-term sequelae in Chernobyl survivors migrated to Israel (Remennick, 2002), as well as those remaining in the FSU (Havenaar et al., 1997). Implications of long-term traumatic sequelae, especially in the complex perimigration context, are discussed. PMID- 15792036 TI - Exposure to human tragedy, empathy, and trauma in ambulance paramedics. AB - Paramedics are exposed to events involving human pain and suffering on a daily basis, many of which are the result of violence perpetrated by 1 individual on another. For the most part, these emergency workers have learned to deal with such events and take them in stride. At times, however, certain circumstances lead workers to develop an emotional connection with the victim or his or her family. When this occurs, paramedics report increased symptoms of traumatic stress. Aspects that can trigger this connection include the victim's alienation from others, profound loss, or the abuse of an innocent child. One of the coping strategies described in these circumstances is to manage the events on a cognitive and technical level while maintaining an emotional distance. Although such a strategy may be protective, it may also have long-term negative effects in terms of interpersonal relationships. This mixed-methods study attempts to better understand factors that lead to higher levels of distress among paramedics within the theoretical framework of emotional and cognitive empathy. PMID- 15792037 TI - Developing and maintaining collaboration in systems of care for children and youths with emotional and behavioral disabilities and their families. AB - Many communities have implemented systems of care in an effort to better coordinate and integrate mental health and other social services for children and youths, while simultaneously managing existing funding sources more effectively. Systems of care represent a fundamentally different way of delivering mental health services and accordingly require new approaches for both developing and sustaining collaboration. This article examines obstacles to collaboration and addresses key factors required to build and sustain collaboration. PMID- 15792038 TI - When is it appropriate to focus on protection in interventions for adolescents? AB - Assertions about the appropriateness of targeting risk or protective factors in interventions for adolescents must be qualified in terms of the outcomes and populations examined in studies and in terms of how risk and protection are measured. The cross-sectional and longitudinal relationships between protection and aggressive behavior found for 388 high-risk adolescents (81.7% male; 50.8% African American/other; average age, 14.5) with serious emotional disturbances in this study validated an intervention focus on protective factors. PMID- 15792039 TI - Behavioral change trajectories of partial hospitalization children. AB - Forecasting treatment outcomes is essential to improving service delivery. Hierarchical linear modeling was used to construct behavioral change trajectories and help predict treatment responsiveness for 97 partial hospitalization children. Overall, children evidenced statistically significant improvement in psychiatric symptomatology. Younger age, female gender, number of previous out-of home placements, and lower IQ scores were associated with increased psychopathology and poorer treatment response. Properly configured day treatment can provide an effective organizing platform for implementing complex treatment regimes. PMID- 15792040 TI - The relationship of parenting stress to adjustment among mothers in prison. AB - Incarcerated mothers at a maximum security prison (N = 362) participated in a study of the relationship between parenting stress, measured by a modification of the R. R. Abidin (1995) Parenting Stress Index, and adjustment, assessed by the L. Derogatis (1993) Brief Symptom Inventory and institutional records of misconduct. Stress associated with limited contact with children was related to higher levels of anxiety, depression, and somatization. Stress concerning visitation was significantly related to anxiety. Stress concerning competence as a parent was associated with elevated anxiety and depressive symptoms as well as with increased institutional misconduct. Incarcerated women experience considerable distress related to parenting, manifest in psychological and behavioral adjustment. PMID- 15792041 TI - Exploring the relationship between child welfare intervention and juvenile corrections involvement. AB - This study of school-age children reported for maltreatment investigated whether child welfare services moderated the risk of later juvenile corrections entry and what factors explained juvenile corrections entry among children who received child welfare services. Non-White children who received in-home child welfare services had a lower risk of juvenile corrections than those receiving no services. Among children who received child welfare services, those also receiving mental health services were at higher risk of juvenile corrections entry. PMID- 15792042 TI - Maternal phenylketonuria: a case study suggesting the use of prenatal psychotherapy to help control phenylalanine levels. AB - Maternal phenylketonuria (MPKU) is an increasingly common concern for health professionals involved in prenatal care. This single-participant study investigated the efficacy of prenatal psychotherapy in improving metabolic control, an important consideration in MPKU pregnancies. Results indicated a robust and positive relationship between psychotherapy and metabolic control. Levels of negative mood and levels of phenylalanine control were also strongly and positively correlated. Implications for treating MPKU are discussed, and prenatal psychotherapy is forwarded as a means of improving birth outcomes in this population. PMID- 15792043 TI - Errorless compliance training: building a cooperative relationship between parents with brain injury and their oppositional children. AB - Errorless compliance training, a noncoercive intervention for improving child compliance, was evaluated in a multiple baseline across-subjects design with 12 oppositional children (ages 2-7 years) and their brain-injured parents, who suffered from cognitive deficits, impulsivity, and/or emotional instability. Generalized and durable increases were observed in child compliance after treatment. Pre/post improvements were also noted on a measure of parent self esteem. Errorless compliance training may be well suited to intervention with parents with cognitive and behavioral impairments that decrease their potential to benefit from traditional parenting approaches. PMID- 15792044 TI - Privatizing medicaid-funded mental health services: trading old political challenges for new ones. AB - States have aggressively pursued privatizing the management of Medicaid-funded mental health services. Although privatized managed care addresses many concerns, it brings several challenges. This article evaluates the impact of privatization on Medicaid-funded mental health services and highlights several contracting issues that should be considered to ensure high-quality mental health care. PMID- 15792045 TI - Health care costs of seriously mentally ill patients enrolled in enhanced treatment. AB - Patients with psychosis (N = 866) were recruited into enhanced or standard Veterans Administration (VA) treatment. Enhanced programs, previously shown to be more effective, were less costly than VA standard care. Adjusted costs fell from $32,000-$55,000 (for the 1st year) to $20,000-$36,000 (for the 4th year). Costs were associated positively with schizophrenia, living in the Northeast region of the United States, and poorer baseline functioning. PMID- 15792046 TI - Bosnian refugees and the stressors of exile: a narrative study. AB - The authors used semistructured interviews to examine exile-related stressors affecting a sample of 28 adult Bosnian refugees in Chicago. The interviews covered 3 areas: life in prewar Bosnia, the journey of exile, and, most centrally, life in Chicago. Primary sources of exile-related distress included social isolation and the loss of community, separation from family members, the loss of important life projects, a lack of environmental mastery, poverty and related stressors such as inadequate housing, and the loss of valued social roles. The implications of these findings for mental health interventions with refugees are considered, and the value of narrative methods in research with refugee communities is discussed. PMID- 15792047 TI - A retrospective study of Arab American mental health clients: trauma and the Iraqi refugees. AB - The purpose of this study was to clarify the mental health needs of Iraqi immigrants who arrived in the United States in the 1990s after the Persian Gulf War. The records of 375 clients were examined at a clinic that serves Arab Americans. More posttraumatic stress disorder and health problems were found in Iraqi refugees than in other clients. Results suggest the need for further research on immigrants with traumatic histories to facilitate effective treatments. PMID- 15792048 TI - The effect of cross-cultural living on personality: assimilation and accommodation among Taiwanese young adults in the United States. AB - Cross-cultural living represents a transition that entails significant psychological disequilibration, to which individuals may respond with assimilation (retention of existing schemas) or accommodation (creation of new schemas; J. Block, 1982). Using the 3 California Psychological Inventory vector scales, the author assessed the personality of 97 Taiwanese young adults studying in the United States prearrival and at 1 and 2 years postarrival. The study found support for both assimilation (stability) and accommodation (change) in personality secondary to cross-cultural living. PMID- 15792049 TI - Journey of family members of homicide victims: a qualitative study of their posthomicide experience. AB - Family members of homicide victims are a neglected population whose posthomicide experience is shaped by the social milieu and primacy of the state's agenda for justice. Themes derived from interviews with 14 families about their lived experience are analyzed and explored through the lens of social constructionist theory, and implications for clinical practice and policy are presented. PMID- 15792050 TI - Predictors of neglect within low-SES families: the importance of substance abuse. AB - A case-control design (N = 203) was used to explore the relative ability of substance abuse, depression, social support, and negative life events to predict neglect status among low-socioeconomic-status families with and without substantiated neglect. Substance abuse emerged as the strongest predictor of neglect status as well as of parental disposition and adequacy of home environment. PMID- 15792051 TI - Gender, perceived parental monitoring, and behavioral adjustment: influences on adolescent alcohol use. AB - This study examined relationships among gender, perceived parental monitoring, externalizing behaviors, and adolescent alcohol use in a 2-wave longitudinal study. Females reported more perceived parental monitoring and less alcohol use than males. Perceived maternal and paternal monitoring were related to less alcohol use over time. Increases in alcohol use were associated with increases in externalizing behaviors over time. Perceived maternal monitoring mediated the relationship between gender and youth drinking and was also a significant predictor of changes in adolescent drinking over time. Perceived paternal monitoring was not a predictor of adolescent drinking in the context of perceived maternal monitoring and externalizing behaviors. The importance of gender in implementation of prevention programs is discussed. PMID- 15792052 TI - The relationship between older and newer parenting categories and child behavior. AB - The relationship between parenting and child behavior was assessed with 15 mothers and 15 fathers of 2- to 3-year-old children. Higher maternal overreactivity was found to be related to more maternal and paternal attributions of child externalizing problems. Self-perception of more paternal nurturance, fewer demands for age-inappropriate responsibility and autonomy, and less laxness were related to fewer paternal attributions of child behavior problems. New constructs for the descriptive categories positive parenting and negative parenting are proposed. These constructs allow one to strongly predict attributions of child behavior. Additionally, each specific behavior forming these new categories is related to specific attributions of child behavior. These specific behaviors and constructs could refine the focus of parent-training programs. PMID- 15792053 TI - Parental emotions following the birth of the first child: gender differences in depressive symptoms. AB - This study addresses psychological issues that put first-time parents at risk for depressive disorders. In a community sample of 107 men and their wives, employment variables, marital quality, and personality attributes (fear of abandonment and loneliness) significantly explained 43% of the variance in men's, and almost 25% in women's, depressive symptoms. Men reported more symptoms than women. Men with heightened fear of loneliness were particularly at risk for depressive symptoms after birth of the first child. PMID- 15792054 TI - Bereavement of spouse caregivers of cancer patients. AB - This study assessed bereavement aspects among 69 widowed spouses (mean age = 61.1; 36.2% men) of deceased cancer patients. They completed questionnaires related to psychological distress as measured by the Brief Symptom Inventory (L. R. Derogatis, 1975a), psychosocial adjustment as measured by the Psychosocial Adjustment to Illness Scale (L. R. Derogatis, 1975b), grief as measured by the Texas Revised Inventory of Grief (T. Faschingbauer, S. Zisook, & R. Devaul, 1987), and communication with spouse (D. H. Olson, D. G. Fournier, & T. M. Druckman, 1982). The findings indicated that women, older people, and past grief were variables that contributed to levels of current distress and grief. The widowed sample as a group showed high levels of distress, suggesting marked vulnerability and the need for planned intervention. PMID- 15792055 TI - Child abuse potential in at-risk African American mothers: the role of life experience variables. AB - The effects of life experience variables on child abuse potential were evaluated in 95 prospectively recruited African American mothers 1 month postpartum. All participants had used opiates or cocaine during pregnancy and were considered high risk for child maltreatment. Abuse potential was assessed with the Child Abuse Potential Inventory, a widely used and psychometrically sound measure with demonstrated predictive validity. Conceptually relevant measures of life experiences and demographic data were obtained from hospital records, maternal self-report, and clinician-administered interviews. Hierarchical multiple regression analysis indicated that exposure to violence in the family of origin and lifetime use of illicit substances made significant independent contributions to the prediction of child physical abuse potential, even after maternal age and current socioeconomic status were controlled for. PMID- 15792056 TI - Prozac and crime: who is the victim? AB - Prozac has been cited in more medication defense criminal cases in the United States than has any other psychotropic drug. In the majority of these cases, defendants are arguing that they are the victims of the drug. Defendants assert that they are victimized by their own involuntary intoxication or that of witnesses and crime victims who have been adversely influenced by Prozac. This article reviews 12 criminal cases in the United States in which Prozac victimization is a salient theme, and it calls for mental health professional organizations to intervene in a growing legal conundrum. PMID- 15792057 TI - The myth of sameness among Latino men and their machismo. AB - This study examined the construct of machismo in relationship to measures of machismo, masculinity, and gender role identity. One hundred forty-eight Latino men with an average age of 36, primarily Mexican American and Puerto Rican, participated. Results indicate that machismo can be characterized as a multidimensional construct, and cluster analyses found that traditional definitions of machismo as authoritarian, emotionally restrictive, and controlling represented only about 10% of the classified Latinos. Most of the sample identified with more emotionally responsive, collaborative, and flexible masculinity styles. Five identity dimensions identified were Contemporary Masculinity, Machismo, Traditional Machismo, Conflicted/Compassionate Machismo, and Contemporary Machismo. Implications include the need to change stereotypes of machismo to be more congruent with the variation in Latino male identity. PMID- 15792059 TI - Disorganized reasoning in Holocaust survivors. AB - In 2 related studies of nonclinical Israeli samples, the long-term sequelae of traumatic Holocaust experiences were investigated from an attachment perspective. In each study, Holocaust survivors were compared with participants who had not experienced the Holocaust, and their attachment style and state of mind with regard to past and present attachment experiences as well as their state of mind regarding unresolved loss were assessed. In both studies, the Holocaust groups were found to be significantly more inclined to show disoriented thought processes around trauma than were the groups without Holocaust background. From an attachment perspective, the authors showed that even after 50 years, traumatic traces of Holocaust experiences are present in the survivors. PMID- 15792058 TI - Nutrition and brain development: social policy implications. AB - Undernutrition among young children is widespread in the United States and has a detrimental impact on brain development. This article explores the risks associated with undernutrition and the potential for recovery when diet and the environment improve. Three policy implications are discussed: (a) increasing access to federal food programs, (b) promoting breastfeeding, and (c) working toward reducing child poverty. PMID- 15792060 TI - Parent-child therapy and maternal projections: tripartite psychotherapy--a new look. AB - Tripartite psychotherapy--where the mother and child are treated together by the therapist--is underutilized as a valuable form of child psychotherapy. Here, a preliminary discussion is based on clinical observations within a brief historical context. Illustrative case examples are offered to exemplify what actually occurs between the mother, child, and therapist in session. This treatment approach is traditionally applied to work with babies and very young children. A basic premise of classical tripartite therapy holds that the presence of the infant in treatment evokes strong transference phenomena in the parent, thus offering direct and immediate intervention within the mother-child relationship. Here, the author stretches the boundaries to extend the age range to older, latency-age children who, within the attachment relationship, may be enacting negative maternal projections. PMID- 15792061 TI - Understanding efficacy in psychotherapy: an ethnomethodological perspective on the therapeutic alliance. AB - In recent years there has been considerable interest in the discovery of a universal paradigm to explain how psychotherapy works. One potential paradigm, known as the therapeutic alliance, is highly correlated with positive patient change. However, the phenomenology and clinical significance of the alliance remains ambiguous. Using a sociological subdiscipline known as ethnomethodology, the author advances a novel perspective on the therapeutic alliance that promises to resolve this ambiguity. PMID- 15792062 TI - Substance use and mental disorder diagnostic profiles in a sample of long-term self-help agency users. AB - This study examined the prevalence of substance use and mental disorder profiles in a sample of mental health self-help attendees. Findings demonstrate that the rates of these disorders are equivalent to or exceed those found in clinical and community samples. Implications for the role that self-help agencies play in the lives of vulnerable individuals, as well as the justification for increased funding from drug and alcohol treatment sources, are discussed. PMID- 15792063 TI - Factors in decisions to seek help from self-help and co-located community mental health agencies. AB - This study examined 673 new users of co-located mental health self-help agencies run by consumers and community mental health agencies to evaluate the relative importance of predisposing, enabling, and need factors in site selection. Although need dominated help site choice, clients' attitudes toward the helpfulness of mental health treatment and their fears of coerced or inadequate care played an important role in setting choice, a choice indicative of a more complex motivational dynamic in help seeking. PMID- 15792064 TI - An empirical typology of youth with severe emotional disturbances. AB - Cluster analysis was used to identify groups of children and adolescents with severe emotional disturbances on the basis of 8 child risk factor questions gathered at intake. The sample of 171 youth was divided into 5 clusters: (a) physically abused, (b) suicide attempt risk, (c) delinquent risk, (d) low risk, and (e) sexually abused. Differences were noted among clusters on a variety of measures of functioning, and cluster groups were different in terms of diagnosis and demographic variables (age, gender, and number of arrests). Youth in Clusters 2 and 3 demonstrated lower functioning on various measures, with Cluster 2 yielding the poorest functioning. Cluster analysis does not supplant diagnosis but may be a useful supplement in identifying meaningful subgroups of youth. PMID- 15792065 TI - Effects of posttraumatic stress disorder and child sexual abuse on self-efficacy development. AB - The symptoms of child sexual abuse and posttraumatic stress disorder (PTSD) affect a child's self-efficacy. A child's self-efficacy beliefs impact the course and treatment of PTSD, because perceived self-efficacy plays a mediating role in children's ability to cope with trauma. Self-efficacy research indicates that emotional competence can be learned and may provide treatment for PTSD that provides symptom reduction as well as a means of substituting problem-solving coping skills for emotion-focused coping skills. PMID- 15792066 TI - Examining the therapeutic utility of restraints and seclusion with children and youth: the role of theory and research in practice. AB - Restraining and secluding children living in residential facilities, though controversial, are commonly practiced by staff to manage disruptive behavior. In an effort to address some of the ongoing issues about the practice, this article examined the reasons for their use, their theoretical underpinnings, and extant empirical evidence in support of the theories. It is suggested that the current array of theories be updated and comprehensive research programs be encouraged to assess their clinical utility with various populations in a variety of settings. PMID- 15792067 TI - Maintaining connections in children's grief narratives in popular film. AB - Children's grief narratives in popular films were examined for their portrayal of connection-maintaining strategies with the deceased. Comparisons were made between strategies found in actual parentally bereaved children and in child characters in films. Implications of these filmed grief narratives for models of grieving and for practice are discussed. PMID- 15792068 TI - Transracial adoptees in the media: 1986--1996. AB - Media reports of interviews with 93 adults transracially adopted as children were content analyzed (percentage agreement = 80%-100%, Cohen's kappas = .61-1.00). Most interviews (63%-88%) involved adoptees who were African American or biracial; in college, graduate school, or employed in professional or managerial occupations; 20-29 years old; and raised in the East or Midwest, with siblings who were the adoptive parents' biological children, other transracial adoptees, or a combination. Forty substantive themes were categorized as adoptive family relationships, direct outcomes, ethnicity and ethnic identification, general adoption issues, and racism experiences. Limitations of media-collected data and the influence of media reports on public opinion and public policies emphasize the importance of scientifically studying controversial phenomena such as transracial adoption. PMID- 15792069 TI - Family ecology and HIV sexual risk behaviors among African American and Puerto Rican adolescent males. AB - This study examined the relationship between family ecology and HIV sexual risk behavior among African American and Puerto Rican adolescent males. Family, psychosocial, and HIV risk factors were assessed in 171 African American and 187 Puerto Rican adolescent males. Findings suggest that family ecology, culture, and gender role variables may differentially affect HIV sexual risk behaviors within these groups. PMID- 15792070 TI - Summary health statistics for U.S. adults: National Health Interview Survey, 2000. AB - OBJECTIVES: This report presents health statistics from the 2000 National Health Interview Survey for the civilian noninstitutionalized adult population, classified by sex, age, race and Hispanic origin, poverty status, and region of residence for chronic condition prevalence, health status and limitations in activity, health care access and utilization, health behaviors, and human immunodeficiency virus (HIV) testing. Also, health statistics by education, income, health insurance coverage, marital status, and place of residence are presented for health status and limitations in activity, health care access and utilization, health behaviors, and knowledge and attitudes toward HIV. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the National Center for Health Statistics, Centers for Disease Control and Prevention, and is representative of the civilian noninstitutionalized U.S. population. Sociodemographic data are collected for everyone in the family during face-to-face interviews with adults present at the time of the interview. The health information for adults in this report was obtained from one randomly selected adult per family. HIGHLIGHTS: In 2000, 64% of adults 18 years of age and over reported excellent or very good health. Eighteen percent of adults under 65 years of age had no health insurance coverage, and 14% of adults did not have a usual place of health care. Eleven percent of adults had ever been told by a doctor or health professional that they had heart disease, and 20% had been told on two or more visits that they had hypertension. Nearly a quarter of adults were current smokers, and 22% were former smokers. Sixty percent did not engage in any leisure-time vigorous physical activity, and, based on estimates of body mass index, 35% were overweight and 21% were obese. PMID- 15792071 TI - Natural causes of death. PMID- 15792072 TI - Beijing morning. PMID- 15792074 TI - Demon, quack, scientist, or saint. Depictions of doctoring in the operatic literature. PMID- 15792073 TI - Cortisone and the burning cross. The story of Percy Julian. PMID- 15792075 TI - Scenes from childhood. PMID- 15792076 TI - We must turn our disease treatment centers into true health centers. PMID- 15792077 TI - Coffee, doughnuts, and mortality. PMID- 15792078 TI - Dead men walking. Reflections on Cotard's syndrome and homelessness. PMID- 15792079 TI - Rectal fumigation. A core rewarming practice from the past. PMID- 15792080 TI - An unlikely muse. The Bellevue Literary Review arises from a legendary city hospital. PMID- 15792081 TI - The physician at the movies: Seabiscuit. PMID- 15792082 TI - The physician at the movies: Master and Commander. PMID- 15792084 TI - Moral leverage won't work! PMID- 15792085 TI - A cheer for the NRMP. PMID- 15792086 TI - Worth the fight. PMID- 15792087 TI - "Spirit" and "soul". PMID- 15792088 TI - More about Fildes. PMID- 15792089 TI - Dog defense. PMID- 15792090 TI - Racial, cultural and ethnic disparities in health and mental health: toward theory and research at community levels. AB - We need to further develop theoretically informed, empirically grounded bodies of knowledge on how community structures, norms, and processes operate in local communities and how they affect human well-being, especially in culturally diverse and ethnic minority communities. Scholars pursuing this agenda will benefit from "population thinking" which focuses attention on human aggregates and on equilibrium processes and social and community mechanisms of change. They will benefit too from considering a promising body of theory and research on social capital. Community level knowledge has profound intervention-related implications at policy levels: such knowledge figured prominently in initiatives associated with 2.5 million residents leaving poverty neighborhoods during the decade 1990-2000. It continues to figure in crucial policy debates and provides a knowledge base on which future federal reports, like 2001s Mental Health: Culture, Race, and Ethnicity-A Supplement to Mental Health: A Report of the Surgeon General, can draw. PMID- 15792091 TI - Community psychology: where do we go from here? AB - Following a developmental analogy, community psychology may be experiencing a "mid-life crisis" as it enters "middle age." The field needs to determine where to go from here. This paper argues that the field should attempt to expand. Expansion can best be accomplished by celebrating the diversity of orientations within the field (e.g., those emphasizing prevention, empowerment, and the ecological perspective) and the wide range of human problems of interest to community psychologists. To promote expansion of the field, community psychologists need to seek out relationships with diverse groups, such as the international community, those working in applied settings, ethnic minorities, and students and early-career professionals. PMID- 15792092 TI - The "big tent" of community psychology: reactions to Paul Toro's 2004 presidential address. AB - This article summarizes discussions that took place following Paul Toro's presidential address for the Society for Community Research and Action (SCRA). Toro argued that community psychology and SCRA are "big tents." He suggested growth through increasing the involvement of ethnic minorities, community practitioners, international colleagues, students and early career professionals and individuals from allied disciplines. Themes in the discussions included: the founding values of community psychology, diversity, and social action. Those present felt that the field was facing a 'mid-life crisis' and needed to re establish its identity. Newer professionals looked to their seasoned colleagues to help define and establish that identity, and involve them in the process. Respondents were supportive of increased involvement of individuals from oppressed groups and practitioners in SCRA. Less attention was paid to increasing the role of international colleagues, professionals from allied disciplines, and members of the community. PMID- 15792093 TI - A tribute to James Gordon Kelly. AB - During the June 2001, eighth biennial meeting of the Society for Community Research and Action in Atlanta, a wide variety of community psychologists across generations attended a tribute in honor of James Gordon Kelly. What follows is an attempt to capture the spirit of the afternoon tribute as expressed through remarks made by colleagues and readings of letters sent by those unable to attend. The wide range of individuals represented here attests to the many additive ways in which Jim has cared about the field of community psychology and has contributed to its essence. Three additional invited contributions are included wherein Dick Reppucci, Rhona Weinstein, and Julian Rappaport reflect on the influence of Jim on their own career and on the development of the field. PMID- 15792094 TI - Role of stressful life events in homelessness: an intragroup analysis. AB - The present paper describes the results of an analysis of individual differences using multi-dimensional strategies-in stressful life event data collected from a representative sample of homeless people (N = 289) in Madrid, Spain, which revealed the existence of three subgroups within the sample. Each subgroup can be defined by the following differentiating characteristics: The first cluster (n = 124) was characterized by economic problems; the second (n = 80) by health problems, alcohol abuse, and death of one or both parents; and the third cluster (n = 50) by an accumulation of stressful life in childhood events and alcohol abuse. In order to define the subgroups, the article also examines the differences found in other variables such as total duration of homelessness, mental and physical health status, and social support availability, among others. The existence of such subgroups within the homeless population emphasizes the importance of designing different interventions for each of these groups, adapted to their diverse needs. PMID- 15792095 TI - A multi-level analysis of community coordinating councils. AB - Coordinating councils are frequently formed to promote a coordinated response to complex social issues (e.g., community health, violence prevention). Little is known, to date, however, about council effectiveness. Prior research on council effectiveness and the correlates of effectiveness have typically relied on case studies. While case studies provide rich descriptive information about the functioning and quality of councils, studies of multiple settings are central to understanding what factors explain variability in perceived effectiveness across councils. Further, the relatively few studies that have examined multiple settings often ignore the multi-level nature of studying councils by failing to disaggregate individual and council level effects. To examine councils' perceived effectiveness in attaining their goals and the correlates of council effectiveness across settings, data were gathered from members (N = 511) and leaders (N = 41) of 43 domestic violence coordinating councils regarding their perceived climate, membership, structure, and accomplishments. Study findings suggest that councils were not perceived as uniformly effective. Councils were more likely to be rated as effective by members and leaders when they were characterized by an inclusive climate and diverse active membership. PMID- 15792096 TI - Contextual competence: multiple manifestations among urban adolescents. AB - The authors develop and validate multidimensional and contextual profiles of competence among low-income, urban, middle adolescents (N = 560). The assessment of contextual competence was based on youth self-reports of involvement, performance, and relationship quality in the peer, school, athletic, employment, religious, and cultural contexts. A principal components analysis of these engagement indices revealed the six expected components with the addition of a component labeled "self-in-context." To identify holistic, multidimensional profiles of contextual competence, scores along the seven domains were cluster analyzed. Nine clusters emerged, each representing a distinct constellation of youth experience. Profiles were associated with demographic variables and youth adjustment. Profiles reflecting high engagement in two or more contexts predicted higher self-esteem and lower depression. In contrast, profiles marked by high engagement in the contexts of athletics or employment predicted more serious delinquency. The authors discuss the implications of these findings for future research and intervention. PMID- 15792097 TI - A tale of two cities: replication of a study on the acculturation and adaptation of immigrant adolescents from the former Soviet Union in a different community context. AB - While a great deal of research has been conducted to understand acculturation and its relationship to adaptation in the new country, surprisingly little attention has been paid to the ways in which the characteristics of the local community impact these processes. The present study addresses this gap in the literature by exploring the potential role of community differences in the acculturation and adaptation processes of 269 refugee and immigrant adolescents from the former Soviet Union who resettled in two different community contexts. Specifically, a prior study on acculturation and adjustment among high school students (D. Birman, E. J. Trickett, & A. Vinokurov, 2002) was replicated with the same emigre population in a contrasting community within the same state. The contrast between these communities allowed us to test hypotheses emerging from an ecological perspective concerning (1) patterns of acculturation, (2) levels of discrimination and its effect on acculturative outcomes, and (3) community differences in the relationship between acculturation and outcomes. In addition to the focus on community differences, the study also employs a multidimensional measure of acculturation and assesses acculturation to both American and Russian culture. Furthermore, adaptation is assessed across different life domains; including peer relationships, family relationships, school adaptation, and psychological adaptation. Findings support the general ecological perspective, suggesting the importance of studying acculturation and adaptation as a reflexive process in which culture and context are very much intertwined. PMID- 15792098 TI - First copulation increases longevity and fecundity of Histiostoma feroniarum (Acari: Astigmata: Acaridida) females. AB - I investigated the influence of insemination at different life stages on female fitness in the mite Histiostoma feroniarum. In this species, males guard immature females at the tritonymph stage to inseminate them immediately after the last moulting. Four groups of females were studied (1) females inseminated 'naturally', i.e. mating occurred immediately after guarding, and then the male was removed (IF/0M), (2) 'naturally' inseminated females, where after insemination the male was replaced by two additional males (IF/2M), (3) virgin females reared without males (VF/0M) and (4) mature, virgin females to which two virgin males were added 3 days after last moulting (VF/2DM). The results show that females inseminated 'naturally' (IF/0M) have higher longevity and fecundity than either virgin (VF/0M) or late-inseminated females (VF/2DM). Furthermore, longevity and fecundity of the former (IF/0M) was also greater than that of females 'naturally' inseminated and subsequently exposed to males (IF/2M). One may suggest that seminal fluids have a positive effect on female fitness. When delayed insemination occurs, such positive effect may not be observed due to a change in features of the sperm access system. Harassment may explain decreased longevity and fecundity of females inseminated 'naturally' compared to females that received additional males. PMID- 15792099 TI - Influence of diet on life table parameters of Iphiseius degenerans (Acari: Phytoseiidae). AB - Abstract. Studies on the reproduction, longevity and life table parameters of Iphiseius degenerans (Berlese) were carried out under laboratory conditions of 25 +/- 1 degree C, 75 +/- 5% RH and 16L:8D h. As food sources for the predatory mite, Ricinus communis L. pollen, all stages of the spider mite Tetrranchus urticae Koch, Frankliniella occidentalis (Pergande) larvae, and Ephestia kuehniella Zeller eggs were selected. All diets were accepted as food by the adult mites. Female longevity ranged from 29.5 to 42.4 days, the highest value was recorded on a diet of Ephestia eggs. The highest percentage of females escaping the experimental arena was observed on the diet consisting of thrips larvae. The highest oviposition rate (1.9 eggs/female.day) was recorded when the predator was fed on spider mites on an artificial substrate. For other diets, oviposition rates ranged from 1.0 to 1.3 eggs/female.day. The intrinsic rate of natural increase (r(m)) of I. degenerans varied between 0.015 and 0.142 females/female x day. The diet consisting of castor bean pollen resulted in the highest population growth whereas the diet on spider mites brushed off onto a bean leaf arena resulted in the slowest population growth. This can be explained by the inability of the predator to cope with the webbing of T. urticae, and the high escape rate of the progeny when reared on spider mites. The percentage of females in the offspring ranged from 40 to 73%. PMID- 15792100 TI - Biology and phenology of the eriophyid mite, Floracarus perrepae, on its native host in Australia, Old World climbing fern, Lygodium microphyllum. AB - The biology and phenology of the eriophyid mite, Floracarus perrepae Knihinicki and Boczek, a potential biological control agent of Lygoditon microphyllun (Cav.) R. Br., was studied in its native range--Queensland, Australia. F. perrepae forms leaf roll galls on the subpinnae of L. mnicrophyllum. It has a simple biology, with females and males produced throughout the year. The population was female biased at 10.5 to 1. The immature development time was 8.9 +/- 0.1 and 7.0 +/- 0.1 days; adult longevity was 30.6 +/- 1.6 and 19.4 +/- 1.2 days and mean fecundity per female was 54.5 +/- 3.2 and 38.5 +/- 1.6 eggs at 21 and 26 degrees C, all respectively. Field studies showed that the mite was active year round, with populations peaking when temperatures were cool and soil moisture levels were highest. Two species of predatory mites, Tarsonemus sp. and a species of Tydeidae, along with the pathogen Hirsutella thompsonii, had significant effects on all life stages of F. perrepae. Despite high levels of predators and the pathogen, F. perrepae caused consistent damage to L. microphyllum at all the field sites over the entire 2 years of the study. PMID- 15792101 TI - Effects of azadirachtin on the two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae). AB - Effects of sublethal azadirachtin exposure to the biological performance of Tetranychius urticae Koch was studied under laboratory conditions. Bioassay was used to asses the effect of different concentrations of azadirachtin on longevity, fecundity, fertility, and offspring development. Azadirachtin (64 and 128 ppm) affected fecundity and mortality but had no effect on fertility and offspring development. A subsequent life-table study with 80 ppm of azadirachtin found that the compound caused a reduction of 50% in survival to adult stage. The peak of reproduction was reached at 5 days causing a decrease in the mean fecundity to almost eight times than of untreated females. The net reproductive rate (R0), the intrinsic rate of increase (r(m)), and the finite rate of increase (lambda) of treated females were lower. Treatment showed a negative value of r(m), resulting in a declining population. These results suggest that azadirachtin could be incorporated in Integrated Pest Management (IPM) programmes of T. urticae. PMID- 15792102 TI - Application of ITS-2 sequences as markers for identification and phylogenetic inference within the genus Geomylichus (Acari: Listrophoridae). AB - Phylogenetic relationships among 18 samples from nine nominal species of Geomylichus (Acari: Listrophoridae), were analyzed based on internal transcribed spacer 2 (ITS 2) sequences, Prolistrophorus sp. was used as an outgroup. Maximum likelihood and neighbor-joining analyses indicated that Geomylichus is formed by at least three internal clades. Genetic distance among Prolistrophorus and Geomylichus was 27.8-35.5%, and among species, populations, and infrapopulations of Geomylichus it was 0.4-21.8%, 0.8-2.4%, and 0.8-1.2%, respectively. Differences between the results of this analysis with conventional taxonomy of the group, as well as the overlapping of genetic distances among species, populations, and infrapopulations, suggest that the definition of species within Geomylichus deserves reconsideration. For example, the recognition of inconsistencies among G. texanus populations associated to different host species, resulting from the molecular analysis, led us to collect new morphological evidence now suggesting that this taxon represents more than one species. PMID- 15792103 TI - Observations on the honey bee tracheal mite Acarapis woodi (Acari: Tarsonemidae) using low-temperature scanning electron microscopy. AB - Observations were made of cryo-preserved honey bee tracheal mites Acarapis woodi (Rennie) using scanning electron microscopy. We describe various new morphological attributes of A. woodi based on the ability of the cryo-technique to capture live mites in natural positions and observe the Low-Temperature Scanning Electron Microscopy (LT-SEM) photographs under a 3-D viewer. Most striking was the observation that each leg has the ability to independently twist its segments with the ambulacrum rotating a minimum of 180 degrees during locomotion; this is a more sophisticated form of locomotion than has been proposed for the Acari. Adult daughter mites are known to be the dispersal instar moving from the tracheal tube to the thoracic hairs of the bee and then transferring to a new bee. We hypothesize that adult tarsal claws and setae on the legs play a role in attachment to hairs during dispersal. However, our evidence is that none of the life stases use their tarsal claws within the tracheal tubes. Larvae were observed to be 'freely moving' within the tracheal system, their tarsal claws rendered inoperative due to an enlarged swollen pulvillar pad. The solenidia of leg I are now known to have striations and the famulus is bifurcated. The bifurcated famulus, solenidial striations, and segmentation of leg IV of females may have taxonomic implications in the family Tarsonemidae. The body and leg setae of adults appear to be used as a tactile tool to sense the amount of space within the tracheal tubes; most of the setae are oriented distally and may help the mite to measure the space or radius of the tracheal tubes. The modified caudal region of the male revealed remnants of the h1 and h2 setae and a smooth clean surface, void of a film, supporting that pharate nymphs are not attached in this species. PMID- 15792104 TI - Ectoparasitic mite Hemipteroseius adleri (Acari: Otopheidomenidae) on the red firebug Pyrrhocoris apterus (Heteroptera: Pyrrhocoridae). PMID- 15792106 TI - Showing some skin. PMID- 15792105 TI - Susceptibility of Amblyomma variegatum ticks to acaricides in Ghana. AB - Abstract. The susceptibility of unfed and fed stages of larvae, nymphs and adult females of Amblyomma variegatum ticks were tested using Shaw's filter paper dip method against four acaricides; chlorfenvinphos and dioxathion, chlorfenvinphos, gamma benzene hexachloride and amitraz at four different concentrations including the recommended dose rates. Based on their lethal concentrations (LC50 & LC90) chlorfenvinphos and dioxathion combined and chlorfenvinphos alone placed first and second, respectively, in all stages except at the unfed nymphal stage where gamma benzene hexachloride topped with a LC50 of 0.001629, while chlorfenvinphos and dioxathion combined and chlorfenvinphos alone had LC50 of 0.001794 and 0.002258, respectively. Amitraz appeared to have a quick knock-down effect on larvae and nymphs but at the recommended dose rate, showed no mortality of the ticks at that stage. However, at a concentration of 0.040%, amitraz showed a 100% inhibition of oviposition and hatching of laid eggs. Gamma benzene hexachloride produced only 66% inhibition of oviposition while chlorfenvinphos and dioxathion combined and chlorfenvinphos alone produced 100% inhibition of oviposition at their recommended dose rates. Fed nymphs were more susceptible than the unfed nymphs. Eggs laid by engorged female ovipositing ticks, applied with gamma benzene hexachloride, hatched. PMID- 15792107 TI - Pediatric dermatology. AB - My goal is that the content of this issue be relevant and useful to your clinical practice, whether it be a clinical pearl in diagnosis, a revelation regarding therapy, or an increased comfort level with dermatologic triage. I hope you have as much fun learning as I had putting this issue together with my respected colleagues. PMID- 15792108 TI - A 4-month-old girl with hemihypertrophy. PMID- 15792109 TI - A 5-year-old girl with a swollen, erythematous foot. PMID- 15792110 TI - Hemangiomas: new insights and classification. AB - Infantile hemangiomas are the most common tumors of infancy, but less common vascular tumors also can affect young infants and children. In most cases, the diagnosis of IH can be made clinically, but imaging studies and even biopsy may be required in less-than-typical cases. With a careful history and physical examination focusing on the timing, location, and type of hemangioma, as well as extracutaneous signs and symptoms, the general pediatrician will know when to be concerned about a high risk vascular tumor and proceed with referral or further evaluations. PMID- 15792111 TI - Tinea capitis: a treatment update. AB - Currently, many experts consider griseofulvin to be the drug of choice for tinea capitis. It is FDA approved for this indication, highly efficacious, and has an excellent long-term safety record. Nonetheless, there is now ample evidence documenting the efficacy and safety of other antifungal agents. Terbinafine, itraconazole, and fluconazole have been used off-label in the United States and United Kingdom for tinea capitis. Several studies have shown that short-term terbinafine, itraconazole, or fluconazole each are comparable in efficacy and safety to griseofulvin. High-dose griseofulvin is still the first-line therapy for tinea capitis in our practice, but a large-scale, multicenter trial of higher dose terbinafine is now ongoing, and positive efficacy and safety results from that study may lead to a change in our standard of care. Terbinafine, itraconazole, or fluconazole currently are used in patients who have either failed griseofulvin or developed adverse reactions to this medication. Families must be informed that these other antifungal agents are not FDA-approved for this indication when they are used. Guidelines for therapy with each of these agents are summarized in Table 5. In addition, the adjuvant use of antifungal shampoos is recommended for all patients in order to decrease the viability of fungal spores present on the hair, as well as for all household contacts to prevent infection or eliminate the carrier state. PMID- 15792112 TI - A treatment approach for atopic dermatitis. PMID- 15792113 TI - How and when to treat molluscum contagiosum and warts in children. AB - Warts and molluscum contagiosum are common skin diseases in children and are usually self-limiting. The decision of whether to treat children with molluscum or warts should be individualized to the patient and his or her family. Considerations include how symptomatic the lesions are, the extent and duration of disease, the ability of the child and the parents to tolerate and comply with treatment recommendations, and any underlying medical conditions (Table, see page 219). Recurrences of molluscum contagiosum and especially warts are common, and realistic expectations regarding the potential for treatment failure and recurrence should be discussed with the child and his or her family prior to initiating any therapy. As pediatric practitioners, we all remain acutely aware of our patients' physical and psychological development and the potential for any intervention to influence this development. Although various treatment modalities now exist for the treatment of these viral diseases, any intervention should be balanced against these considerations. PMID- 15792114 TI - Connective tissue disease in children. AB - As our understanding of connective tissue disease expands, so too does our therapeutic armamentarium. We have learned that autoimmunity triggers inflammation through unchecked, proliferative cell-mediated inflammation. By targeting this arm of the cytokine cascade, it may be possible to arrest further progression. Several biologic agents, such as etanercept, alefacept, infliximab, efaluzimab, and, recently, adalimumab, have come to market for adult psoriasis and are now undergoing trials for juvenile SLE, psoriasis, and psoriatic arthritis. Of note, etanercept has been used successfully in juvenile rheumatoid arthritis for more than 10 years. These agents target cell-mediated inflammation through various mechanisms and hold great promise for the treatment of many of the disease states discussed above. Moreover, the biologics carry an improved side-effect profile not seen with traditional agents such as corticosteroids and will be central in the evolution of targeted therapeutics for these complex immunologic diseases. PMID- 15792115 TI - Future directions in pediatric dermatology. PMID- 15792116 TI - FAS and FAS-L expression by tumor cells and lymphocytes in breast carcinomas and their lymph node metastases. AB - FAS receptor (FAS, CD95) and FAS ligand (FAS-L, CD95-L) are complementary members of a particular apoptotic pathway that plays a major role in immune regulation. The activation of FAS-L may trigger cytotoxic mechanisms leading to the death of FAS-expressing cells. Tumor cells and tumor-infiltrating lymphocytes (TIL) may express FAS and FAS-L in various proportions, and their interplay may affect tumor behavior. In the present study, we explored the expression of FAS and FAS-L in 28 mammary carcinomas (19 ductal and 9 lobular) and in their lymph node metastases. The expression of these mediators in immunostained sections was graded and evaluated comparatively between normal and neoplastic mammary epithelium, between tumor cells and TILs, and between mammary carcinoma cells and their lymph node metastases. We demonstrated the coexpression of FAS and FAS-L by breast carcinoma cells and TIL, with FAS expressed more strongly by normal epithelial cells and TIL than tumor cells. FAS-L was better stained on tumor cells than on TIL. There was equal or greater expression of FAS and FAS-L in the primary tumors and their TIL than in the metastatic counterparts. Comparing the expression of FAS with that of FAS-L, we recorded FAS equal or stronger than FAS L in the primary mammary tumors and the reversal of their expression, FAS-L greater than FAS in the lymph node metastases. These results are consistent with reports of studies with other tumors, suggesting that the upregulated FAS-L indicates an increased ability of tumor cells to induce apoptosis in TIL and in the normal tissues invaded. However, it is understood that the FAS/FAS-L system, although essential for apoptosis, is only a contributing factor to the complex process of tumor invasion and antitumor defense. PMID- 15792117 TI - Expression analysis of cystatin C and M in laser-capture microdissectioned human breast cancer cells--a preliminary study. AB - Cathepsins B and L, implicated in the progression of malignant tumors, are regulated by a family of endogenous inhibitors referred to as the cystatins. Cystatin M was identified by differential display as down-regulated gene in metastatic breast cancer cells. However, this finding has yet to be confirmed in clinical breast cancer specimens. Our objective is to examine the expression levels of cystatins C, M, and cathepsins B and L mRNA in breast cancer cells isolated by laser capture microdissection. The mRNA and protein levels of cathepsin B, L, and cystatin C and M in breast cancer specimens were determined utilizing laser capture microdissection/RT-PCR, Western blotting, and immunohistochemical methods. Expression levels of either cystatin M or C were not significantly different between lymph node-positive and -negative breast carcinomas. Increased expression levels of both cystatin M and C correlated significantly with larger tumor size. Cystatin M mRNA was detected by in situ hybridization in both primary and metastatic breast cancer cells. Our findings are at variance with a previous report proposing a metastasis suppressive function for cystatin M. Therefore, additional studies in a larger series with adequate follow-up are necessary to elucidate the biologic significance of cystatin M expression in breast cancer metastasis. PMID- 15792118 TI - Combined application of RT-PCR and immunohistochemistry on paraffin embedded sentinel lymph nodes of prostate cancer patients. AB - The detection of tumor cells in the sentinel lymph node (SLN) is of great importance for the prognosis of cancer patients. At present, immunohistochemistry and RT-PCR for tumor marker expression are the most sensitive techniques available for this analysis. However, so far, most RT-PCR-based analyses of SLNs have been performed on fresh material, excluding a direct comparison with the (immuno)histologic results. In our view, this does not entirely aid routine diagnosis. We established an efficient method for RNA extraction and RT-PCR from paraffin sections of SLNs from prostate cancer patients and compared the results with the (immuno)histologic data of adjacent sections. Amplifiable RNA was obtained from 133 SLNs of 68 prostate cancer patients. Correlation of PSA specific RT-PCR with (immuno)histologic findings showed a positive and negative predictive value of 83% and 100%, respectively, for the prostate cancer patients investigated. Four of 12 patients with biochemical relapse, but without (immuno)histologically detectable tumor cells were RT-PCR-positive for PSA. We found that single sections of paraffin-embedded SLNs are suitable for routinely performed RT-PCR. Combined with (immuno)histology, PSA-specific RT-PCR is a revealing supplementary technique for the detection of tumor cells in SLNs of prostate cancer patients. PMID- 15792119 TI - Usefulness of immunohistochemistry for recognizing metastatic colorectal adenocarcinoma in infarcted lymph nodes. AB - Lymph node infarction is a spontaneous coagulative necrosis of the affected lymph node and is frequently associated with concurrent and subsequent malignant lymphoma. However, this phenomenon appears to be rarely associated with metastatic carcinomas. Here, we report on the histopathologic and immunohistologic findings of three cases showing lymph node infarction in the regional lymph node associated with metastatic colorectal adenocarcinoma. Histologically, coagulative necrosis of metastatic carcinoma was surrounded by a thick rim of granuloma consisting of histiocytes with or without epithelioid features, foamy cells, and a small number of lymphocytes. The immunohistochemical study of the coagulative necrosis demonstrated that cytokeratins (AEI/AE3 and CAM5.2) and carcinoembryonic antigen (CEA) were well preserved in all three cases. However, compared with viable tumor tissues, only a few tumor cells were positive for epithelial membrane antigen. Using formalin-fixed and paraffin embedded tissues, immunostaining for cytokeratins and CEA of the lymph node containing necrotic carcinoma may provide clinically valuable information. PMID- 15792120 TI - Correlation of donor-derived keratinocytes and severity of graft-versus-host disease (GVHD) in epidermis. AB - Recent reports suggest that bone marrow might serve as a source of skin progenitor cells. Graft-versus-host disease (GVHD) is a systemic disease that involves the skin, the liver, and the gastrointestinal tract, and contributes to transplant-related morbidity and mortality. To evaluate whether donor-derived bone marrow cells participated in its pathophysiology, we correlated the severity of GVHD in skin from sex-mismatched recipients with the percentage of donor derived keratinocytes using fluorescence in situ hybridization (FISH) for detecting the Y chromosome. Y-positive signals were observed in female epidermis in both keratinocytes and lymphocytes. These results support the notion that donor-derived hematopoietic stem cells may contain pluripotent stem cells. Furthermore, there was a strong correlation between the frequency of Y-positive keratinocytes and the histopathologic grade of GVHD (p = 0.004), as well as with the number of Y-positive lymphocytes infiltrating the epidermis (p = 0.005), suggesting a role for donor-derived keratinocytes in the pathophysiology of GVHD. However, no clusters of Y-positive keratinocytes were found in the epidermis, and no correlation was observed between the number of Y-positive keratinocytes and time course of GVHD after transplantation. We hypothesize that donor-derived stem cells may play a role in the regeneration of damaged keratinocytes in GVHD. PMID- 15792121 TI - Loss of maspin is a helpful prognosticator in colorectal cancer: a tissue microarray analysis. AB - Mammary serpin (Maspin) belongs to the serine protease inhibitor (serpin) superfamily and has been identified as a tumor suppressor. In addition, a p53 dependent regulatory pathway of maspin in human cancer has been indicated. The role of maspin in the tumorigenesis and progression of colorectal cancer is still unclear. Therefore, the first aim of the present study was to determine the prognostic value of maspin protein expression for the recurrence-free and overall survival of patients with colorectal carcinoma undergoing left- or right-sided colectomy. Secondly, maspin expression was correlated with p53 protein expression to gain additional information about a possible regulatory influence of the wild type p53 protein on maspin; it was also correlated with further patient and tumor characteristics (age, sex, TNM, disease stage, tumor localization, and grading). An immunohistochemical study was performed on 280 carcinoma specimens using the tissue microarray technique. In addition, 80 colorectal adenomas and 60 tumor free tissues were investigated. Maspin protein expression was detectable in 88 100% of the adenomas and non-tumorous tissues and in 193 out of 280 carcinoma patients (69%; maspin-positive). After a median follow-up of 102 months (23-140 months), the median recurrence-free survival was 80 months for maspin-positive cases (M +) and 42 months for maspin-negative cases (M-) (p = 0.02). The median long-term survival was 98 months for M + and 57 months for M- (p = 0.03). After 5 years, M + and M- patients had a total survival of 69% and 38%, and, after 10 years, 45% and 9%, respectively. Mutant type p53 expression was detectable in 178 colorectal carcinomas (64%). Mt p53 was positive in 91 out of 193 M + (47%) compared with 87 of 87 M- (100%, p<0.001). This study showed that loss of maspin protein correlates with p53 protein activity, with a higher likelihood for the development of tumor relapse, and with a decreased recurrence-free and overall survival in colorectal carcinomas. The determination of the immunohistochemical expression status of maspin might be a helpful independent prognosticator and an applicable tool for the development of therapeutic strategies for patients with this disease. PMID- 15792122 TI - P53, bcl-2 and Ki-67 expression in adenoid cystic carcinoma of the palate. A clinico-pathologic study of 21 cases with long-term follow-up. AB - Adenoid cystic carcinoma (ACC) is an indolent tumor that pursues a protracted clinical course with recurrences and late metastases. The aim of this study was to investigate immunohistochemically the expression of p53, bcl-2 protein, and Ki 67 in 21 cases of ACC of the palate, all with a minimum of 10 years and a maximum of 22 years of clinical follow-up. These results were also analyzed with regard to different clinical prognoses of the histologic subtypes of ACC. High expression of p53 and bcl-2 was noted in 19 out of 21 ACC cases (90%), in which most tumor cells (from 66% to 99%) proved to be immunopositive. A relation to the histologic types, clinical staging, and survival was not found. Therefore, the high immunoreactivity against these oncoproteins in the same tumor cells suggests that these two oncogenes may be involved since the early stage of carcinogenesis. Loss of function of the p53 protein combined with bcl-2 upregulation might give the tumor cells a double growth advantage, because uncontrolled proliferation is combined with a reduced cell death rate. The interaction with other oncogenes may then trigger a multistep process able to promote tumor progression. The low labeling index Ki-67 was detected in nine out of 21 cases (42%), with a low percentage of tumor cells (from 3% to 15%) being positive, whereas the remaining 12 cases were negative. We found no relation to the histologic types, clinical staging, and survival; however, the low proliferation rate could explain the natural course of tumor. PMID- 15792123 TI - Does Cytokeratin7/20 immunoreactivity help to distinguish Barrett's esophagus from gastric intestinal metaplasia? Results of a prospective study of 75 patients. AB - Barrett's esophagus is a recognized risk factor for the development of esophageal dysplasia and carcinoma. Unfortunately, gastric incomplete intestinal metaplasia arising in Short Segment Barrett's esophagus can be indistinguishable histologically on hematoxylin/eosin stains. Distinct patterns of CK 7 and CK 20 immunohistochemical expression have been demonstrated to be both highly sensitive and specific for Barrett's esophagus, but have not been found in gastric metaplasia. The aim of our study was to test whether immunostaining with CK 7/20 helps to distinguish between Barrett's epithelium and gastric incomplete metaplasia. Cases of long segment Barrett's esophagus, short segment Barrett's esophagus, and cases with a normal gastroesophageal junction, as well as specimens with gastric antral intestninal metaplasia, were examined: three patterns were defined. Barrett's pattern (superficial CK 20 staining; superficial and crypt CK 7 staining); gastric pattern (superficial and crypt staining of both markers); other patterns (different from Barrett and gastric types). Seventy-five patients were enrolled in this study, 26 with long segment Barrett's esophagus, 21 with short segment esophagus, 13 with intestinal metaplasia of the cardia, and 18 with antral intestinal metaplasia. The Barrett pattern showed a high specificity of 97%, but a sensitivity of only 30% in patients with short segment Barrett esophagus. Our results do not confirm the hypothesis that CK 7/20 immunostaining can be used for a reliable differentiation between incomplete intestinal metaplasia and Barrett's epithelium. PMID- 15792124 TI - Bone marrow fat-cell enlargement in early steroid-induced osteonecrosis--a histomorphometric study of autopsy cases. AB - Some animal studies and magnetic resonance imaging studies suggest that there may exist a relationship between abnormal lipid metabolisms and osteonecrosis. The purpose of this study was to examine the size of bone marrow fat cells in the early osteonecrosis femoral head using autopsy specimens. We compared the size of bone marrow fat cells in the viable areas in the following three autopsy groups: the early osteonecrosis group (4 femoral heads); the steroid-administered group (without osteonecrosis) (n = 10), and the normal group (n = 19). In addition, after adjusting for age and sex, the size of bone marrow fat cells was compared using multiple regression analysis. The size of bone marrow fat cells was significantly larger in the early osteonecrosis group (84.7+/-5.5 microm) than in both the steroid-administered group (75.3+/-4.3 microm) and the normal group (76.3+/-4.9 microm) (p<0.01 and p<0.05, respectively). After adjusting for age and sex, the size of bone marrow fat cells in the early osteonecrosis group was significantly larger as compared with the other groups. This study suggests that in steroid-induced osteonecrosis, the size of bone marrow fat cells increases significantly at an early stage. PMID- 15792125 TI - Diffuse pulmonary hamartoma: a case report. AB - Pulmonary hamartoma usually occurs as a benign, well-circumscribed single nodule in the lung parenchyma. We report on a unique case of hamartoma that extended along the bronchial tree, formed endobronchial polypoid lesions, and expanded into the lung parenchyma. The patient, a 48-year-old man, was admitted with dyspnea and chest pain. A transbronchial biopsy was performed on a tumorous lesion that was diagnosed histologically as a hamartoma. As this lesion was found to be growing diffusely along the bronchial tree, a left pneumonectomy was performed. Gross examination showed that yellowish soft tissue had surrounded the bronchial tree and extended into the lung parenchyma. Histologically, the lesion contained mainly mature adipose tissue, cartilage, and muscle tissue with a minor component of short spindle cells in a myxomatous matrix. The patient was diagnosed as having diffuse pulmonary hamartoma. PMID- 15792126 TI - Chief cell hyperplasia with structural and nuclear atypia: a variant of fundic gland polyp. AB - A case of an unusual variant of fundic gland polyp (FGP) composed of chief cell hyperplasia with structural and nuclear atypia in an 87-year-old woman is presented. Gastrointestinal endoscopy revealed a sessile polyp in the cardia/ corpus transition zone and a polypoid lesion in the fundus. Histologically, the polyp in the cardia/corpus showed a typical appearance of FGP, while that in fundus demonstrated a tumorous lesion composed of irregular branched tubules with nuclear stratification. Despite the structural distortion and nuclear atypia, mitotic figures were absent and MIB-1 positive cells were less than 3%. Immunohistochemically, the cytoplasms of the tubules were negative for gastric mucin and Muc-5AC glycoprotein, but mostly positive for pepsinogen-I, indicating that the proliferated glands consisted mainly of chief cells, not mucous cells. Parietal cells were occasionally found in the glands. At the periphery of the lesion, microcysts composed of parietal cells, chief cells, and mucous cells had developed. Altogether, the polyp in the fundus was diagnosed as an unusual variant of FGP with chief cell hyperplasia. This FGP should be differentiated from tubular adenocarcinoma. Proliferation of chief cells with occasional parietal cells is critical for the differential diagnosis. PMID- 15792127 TI - Composite glandular-endocrine cell carcinoma of the stomach. Report of two cases with goblet cell carcinoid component. AB - Composite glandular-endocrine cell carcinoma (CGECC) is recognized as a special type of gastric tumor composed of ordinary adenocarcinoma and neuroendocrine tumors. Goblet cell carcinoid (GCC) is a well-established type of appendiceal carcinoid, but the GCC component has not been well delineated in CGECC of the stomach. We report on two gastric CGECCs with a GCC component, analyzing the histologic components by immunohistochemistry. On initial biopsy, both cases were diagnosed as signet-ring cell carcinoma. However, the resected tumors consisted of three components: signet-ring cell carcinoma, GCC, and glandular adenocarcinoma. Although some signet-ring carcinoma cells and goblet carcinoid cells were indistinguishable by hematoxylin and eosin staining, E-cadherin immunostaining disclosed a definitive difference regarding the staining pattern in these cells. Both patients are well, with no recurrent tumor for about 10 years of follow-up. CGECC with a GCC component may have been confused with conventional adenocarcinoma with signet-ring cells. In cases of advanced signet ring cell carcinoma with good prognosis, the possibility of such CGECC has to be considered. PMID- 15792128 TI - Ductal carcinoma in situ-like structures in metastatic breast carcinoma. AB - In this report, we present two cases of axillary lymph node metastatic breast carcinoma with features mimicking ductal carcinoma in situ (DCIS): one was of the comedo-like type and the other was suggestive of the micropapillary type. In the first case, the primary tumor presented DCIS of the comedo type; however, in the second case, the primary tumor consisted only of the invasive ductal component. Immunohistochemistry against smooth muscle actin, S100-protein, CK5/6, CD10, P63, and 34betaE12 did not identify myoepithelial cells either in DICS of the first primary tumor or in both metastases. These features probably do not represent the true DCISs, but only mimic them. This observation suggests that a proportion of "primary DCIS" may constitute an invasive pseudo-DCIS carcinoma, and immunohistochemical identification of myoepithelial cells may be helpful in such cases. PMID- 15792129 TI - Myxoma of the renal capsule. AB - Myxomas are uncommon soft-tissue neoplasms, which are extremely rare in the kidney, with only five cases documented in the intraparenchymal location. However, renal capsular myxoma has not yet been reported. We describe a unique case of a clinically detected renal myxoma arising in the capsule. A 37-year-old man receiving treatment for epididymitis sought medical assistance for infertility. A radiological examination incidentally discovered a right renal tumor. The mass intruded into the perirenal tissue and measured 6 cm in major diameter. The resected kidney contained a well-circumscribed gelatinous capsular tumor. It was composed of sparse, bland, slender, spindle-shaped cells scattered in large amounts of basophilic interstitial mucoid material. The tumor cells showed diffuse immunoreactivity for vimentin. Occasional cells stained for alpha smooth muscle actin and calponin. Reactivity was negative for S100 protein, epithelial membrane antigen, pancytokeratin, neurofilament protein, and h caldesmon antibodies. Ultrastructural examination revealed fibroblast-like cells with long thin cytoplasmic processes, prominent rough endoplasmic reticulum, a well-developed Golgi complex, and secretory vesicles. No basal lamina was identified around the tumor cells. The differential diagnosis includes many other benign and malignant soft-tissue lesions exhibiting prominent secondary myxoid features. It is important to consider a renal capsular myxoma when examining lesions at this anatomic site to avoid misdiagnoses and to ensure that the patient receives appropriate treatment and prognostic information. PMID- 15792130 TI - Maxillary African histoplasmosis: unusual diagnostic problems of an unusual presentation. AB - Among bone lesions of African histoplasmosis, those affecting the jaw are relatively rare and concern, with other facial involvements, particularly infants and adolescent patients with an usual uncompromised immunologic status. As clinical and radiologic features are not specific, the differential diagnosis to other mandibular diseases is difficult. We report on a case of African histoplasmosis that involved the right mandibula of a 17-year-old Congolese boy with a persistent and fungiform cutaneous ulceration. As mycologic tests had not been carried out initially, the disease was histologically diagnosed on the basis of the presence of numerous intra-cytoplasmic large yeasts in a granulomatous lesion containing giant cells. As it is impossible to confront the histologic diagnosis with mycologic tests in such a situation, the problems of the differential diagnosis to other deep fungus infections and to some yeast-like foreign body-granulomas encountered at the microscopical level underline the importance of culturing organisms from lesions to confirm the histologic diagnosis. It is worth considering this pathology at least for three reasons: it usually mimicks a malignant jaw tumor; it may constitute a migrant pathology; and prognosis is commonly favorable with amphotericin B treatment. PMID- 15792131 TI - Unusual recurrence in ureters and maxillary sinus of Langerhans cell histiocytosis involving mastoid bone. AB - We present a case of Langerhans cell histiocytosis (LCH) diagnosed in the mastoid bone. The tumor recurred in the ureter and maxillary sinus mucosa two years later. The diagnosis of LCH was based on morphology and immunohistochemistry. Involvement of the ureter and the maxillary sinus in LCH is extremely rare. To the best of our knowledge, this is the first case of LCH affecting the mastoid bone in a 16-year-old boy and recurring later in the ureter and maxillary sinus mucosa. PMID- 15792132 TI - Renal epithelioid angiomyolipoma--a close mimic of renal cell carcinoma. Report of a case and review of the literature. AB - The epithelioid variant of angiomyolipoma (EAML) is a rare tumor of unpredictable behavior that is composed of epithelioid, spindle, and giant cells and contains no or only a minimal amount of lipomatous tissue. The picture can lead to an erroneous diagnosis of renal cell carcinoma or sarcoma. We report on a case of EAML in the kidney of a 47-year-old female without any signs of tuberous sclerosis and review the literature. Grossly, a well-demarcated, spheroid, largely hemorrhagic tumor measuring 4.2 cm in diameter occupied the central third of the kidney. Histologically, it was solid, highly cellular, with occasional microcysts, composed of medium to large epithelioid cells with clear or oxyphilic cytoplasm, short spindle cells, and numerous giant multinucleated cells. After extensive sampling, adult-appearing fat tissue was found to present as rare foci of microscopic dimensions. Immunohistochemically, the tumor cells showed positive reactions with antibodies against HMB-45, melan A, CD-68, muscle-specific actin, and, rarely, smooth muscle actin. Cytokeratins and epithelial membrane antigen were negative. The EAML is a variant growing in a carcinoma-like pattern that can lead to an erroneous diagnosis of renal cell carcinoma. An extensive sampling and HMB-45 and CD-68 positivity combined with cytokeratin negativity are of paramount importance for the correct diagnosis. As a sporadic renal tumor it followed a benign course in most of the reported cases. PMID- 15792133 TI - Renal variant of Anderson-Fabry disease and bilateral renal cell carcinoma. AB - Anderson-Fabry disease (AFd) is an X-linked metabolic disease with clinical manifestations secondary to accumulation of glycosphingolipids in various tissues. We report the first case in which a patient with renal variant of AFd and chronic renal failure developed bilateral conventional renal cell carcinoma. His metabolic disorder was diagnosed only after histopathologic study of the kidney specimen resected because of the tumoral lesion. There is no clear etiologic relation between the metabolic and neoplastic disease. As AFd is not common or well known and its clinical manifestations tend to be nonspecific, the disorder is often unrecognized, misdiagnosed, or diagnosed late in life. The pathologist should be aware of this disorder when evaluating a kidney specimen from patients with chronic renal failure of unknown cause. PMID- 15792134 TI - Intramyofiber erythrocytes in skeletal muscle. AB - The authors describe the serendipitous discovery of clustered erythrocytes within myocytes in a biopsy obtained during repair of a torn tendon. Their perusal of the literature failed to uncover an analogous observation. PMID- 15792135 TI - Benign mixed epithelial and stromal tumor of the kidney (MEST) with cytogenetic alteration. AB - Benign mixed epithelial and stromal tumor of the kidney (MEST) is a new, rare entity. These tumors are composed of two components: a stromal and an epithelial one. Clinical outcome is usually good, no specific cytogenetic alterations have been described up to now. We describe for the first time a case with translocation t (1; 19). PMID- 15792136 TI - Hyperacute rejection in heart allografts: case studies by J. Kemnitz et al. (Pathol. Res. Pract. 187: 23-29, 1991). PMID- 15792137 TI - [Non-ischemic neurological manifestations in patients with primary antiphospholipid syndrome]. AB - Ischemic disorders of brain blood circulation caused by brain artery thrombosis due to antiphospholipids-induced anticoagulopathy are main neurological appearances of primary antiphospholipid syndrome (PAPS). A number of neurological disorders in patients with PAPS are the result of primary involvement of the brain and peripheral nervous system. We analyzed the spectrum of neurological non ischemic PAPS manifestations in 125 patients (102 female, 23 male, mean age--37.5 +/- 11.3 years) with definite PAPS. These manifestations included headache (67%), epileptic seizures (23%), chorea (15%), optic neuropathy (9%), peripheral neuropathy (6%), multiple sclerosis like syndrome (MSLS) (8%), acute psychosis (2%), myasthenic syndrome (1%), non-vascular parkinsonism (1%). In the development of non-ischemic PAPS manifestations, antiphospholipids as well as other antibodies produced as a result of immune disregulation (antibodies to acetylcholine receptors in myasthenic syndrome, antineuronal antibodies in MSLS) may have pathogenic significance. In some cases a role of infection involved in PAPS manifestation cannot be ruled out. PMID- 15792138 TI - [Autonomic disorders in dementia with Lewy bodies]. AB - Fifty patients diagnosed with dementia with Lewy bodies (DLB) according to criteria of I. MacKleith et al (2000), mean age 71.1 +/- 6.1 years, have been studied. Severity of dimentia was evaluated using a battery of neuropsychological scales and tests. A state of autonomic nervous system function was determined during clinical examination, the results of which were used as a basis for the Scale of autonomic disorders severity, and by instrumental methods. Most of patients with DLB had autonomic disorders, such as orthostatic hypotension, heat intolerability, dryness of cutaneous epithelium and mouth mucous membranes, urination disorders. The study of reactivity of autonomic nervous system revealed a predominance of parasympathetic reactions. The authors believe that autonomic disorders are typical for DLB and are the result of affection of the peripheral nerves, stem and spinal autonomic structures. PMID- 15792139 TI - [Psychopathology of self-consciousness in acute alcoholic hallucinosis]. AB - To study self-consciousness changes in initial, manifesting and final stages of acute alcoholic hallucinosis, 60 patients have been examined. In 51 of them, the disease developed on the background of pseudo-dipsomania or constant hard drinking and in 9--of abstinent state. Initial stage of acute alcoholic hallucinosis was characterized by negative type of self-consciousness reaction. In manifesting stage the following variants of self-consciousness were detected: disturbation (vital, somatophysical, autopsychic), suppression (harmonic, dissociated), depersonalization and hyperactivity. In final stage, there were disactualization, nosognosia (adequate-, hypo- and hypernosognosia, anosognosia), partial and total reintegration of self-consciousness. These disorders of self consciousness appear to be of prognostic and diagnostic value. PMID- 15792140 TI - [Semax in prevention of disease progress and development of exacerbations in patients with cerebrovascular insufficiency]. AB - One hundred and eighty-seven patients with different stages of cerebrovascular insufficiency (CI) have been examined. A diagnosis of CI was based on the results of neurological and neuropsychological study, ultrasonic dopplerography, rheo- and encephalography, electrocardiography, brain MRI and eyegrounds examination. Neurological scales were used for neurological status assessment and further data processing. The study aimed at evaluation of tolerability and clinical efficacy of the medication and complications in CI course. Semax treatment resulted in significant clinical improvement, stabilization of the disease progress and reduced a risk of stroke and transitory ischemic attacks in the disease course. The drug is featured by minor percent of side-effects and is well tolerated by patients, including those of older age groups. PMID- 15792141 TI - [An effect of antioxidant drugs on symptoms of sensomotor polyneuropathy and affective disorders in patients with diabetes mellitus]. AB - An effect of 3-oxypiridine and succinic acid derivates--emoxipine, reamberin and mexidole--on severity of clinical symptoms of distal sensomotor polyneuropathy, anxiety and depression was studied in 120 patients with diabetes mellitus with syndrome of diabetic foot. These medications were used in addition to basic antidiabetic therapy. After 14 days of the treatment an equally pronounced decrease of integral index of "negative" neurological symptoms and reduction of anxiety-depressive disorders were detected. PMID- 15792142 TI - [Efficacy of pronoran in age-related memory impairment]. AB - Thirty patients aged 45-62 years (mean 54.8 +/- 3.7) have been studied. Pronoran (piribedil) was used for treatment of age-related memory impairment in dosage 50 mg per day during 30 and 90 days (2 groups of 15 patients). Psychophysiological examination included a study of sensomotor activity (time for visual-motor reactions--simple and complex) and functional lability of visual analyzer as well as evaluation of cognitive processes--memory and attention. Patients were examined before and after therapy of different duration. Pronoran exerted a positive effect on psychophysiological state of elderly people, improving memory and attention and increasing the velocity of psychomotor reactions and lability of nervous processes. The longer was the therapy, the more improved were above traits. Along with cogniotropic and mnemotoric (an influence on operative memory) effects of pronoran, its non-specific activating impact is described. PMID- 15792143 TI - [Clinical, tomographic and immunogenetic study of patients with infantile cerebral palsy]. AB - A neurovisual and immunogenetic study of patients with different forms of cerebral palsy was conducted. Morphological peculiarities of each form were described. A frequent combination of pathology of cerebrospinal fluid spaces and periventricular area with disruption of neuronal migration and development of brain mass and volume was found. HLA-typing revealed a significant association of the disease with antigen B13. An association of cerebral palsy with particular genetically determined vulnerability of fetal brain to lesions disrupting genetic program for neuroontogenesis is suggested. PMID- 15792144 TI - [Neuroendocrine aspect of women epilepsy in last two decades publications]. PMID- 15792145 TI - [An experience of topamax usage as monotherapy in different forms of epilepsy in outpatient clinics]. PMID- 15792146 TI - [An effect of iodine deficiency on neuromental development: a modern view on the problem]. PMID- 15792147 TI - [Immune lesions of the nervous system in some forms of rheumatic disorders]. PMID- 15792148 TI - [A new quality of the therapy of anxiety and depression--escitalopram]. PMID- 15792149 TI - [Epidemiology and war: time for substantive humanitarian action]. PMID- 15792150 TI - [Waste disposal, triangles and 'hot' areas]. PMID- 15792151 TI - [Environment and health for children]. PMID- 15792152 TI - [The hot summer of Campania is not yet over]. PMID- 15792153 TI - [Cause-specific mortality in an area of Campania with numerous waste disposal sites]. AB - OBJECTIVE: To investigate cause-specific mortality in an area of Campania region, in the surroundings of Naples, characterized by many toxic waste dumping grounds sites and by widespread burning of urban wastes. DESIGN: The study area was characterized by examining the spatial distribution of waste disposal sites and toxic waste dumping grounds, using a geographic information system (GIS). Mortality (1986-2000) was studied in the three municipalities of Giugliano in Campania, Qualiano and Villaricca, encompassing a population of about 150,000 inhabitants. Mortality rates of the population resident in the Campania region were used in order to generate expected figures. OUTCOMES: Causes of death of a priori interest where those previously associated to residence in the neighbourhood of (toxic) waste sites, including lung cancer, bladder cancer, leukemia and liver cancer. RESULTS: Overall 39 waste sites, 27 of which characterized by the likely presence of toxic wastes, were identified in the area of interest. A good agreement was found between two independent surveys of the Regional Environmental Protection Agency and of the environmentalist association Legambiente. Cancer mortality was significantly increased, with special reference to malignant neoplasm of lung, pleura, larynx, bladder, liver and brain. Circulatory diseases were also significantly in excess and diabetes showed some increases. CONCLUSIONS: Mortality statistics provide preliminary evidence of the disease load in the area. Mapping waste dumping grounds provides information for defining high risk areas. Improvements in exposure assessment together with the use of a range of health data (hospital discharge cards, malformation notifications, observations of general practitioners) will contribute to second generation studies aimed at inferring causal relationships. PMID- 15792154 TI - [Time trends in infectious disease mortality in Italy: 1969-1999]. AB - OBJECTIVE: To describe the global impact of infectious diseases on mortality in Italy from 1969 to 1999. DESIGN: Statistical analysis of routinely collected mortality data, using a revised definition of infectious causes of death based on target organs. SETTING: The present paper summarizes time trends of infectious disease mortality widely discussed in the Atlas "30 Anni di Malattie Infettive in Italia: Atlante di Mortalita". MAIN OUTCOME MEASURES: Age standardized mortality rates (/100,000); standardized mortality ratios (SMR); percentage of deaths attributable to infectious diseases. RESULTS: Apart from HIV infection and AIDS, infectious diseases were responsible of 1.7% of the overall mortality that occurred in Italy in the study period: 57.5% of such deaths were not included in the ICD8 and ICD9 codes for infectious diseases. The mortality for all infectious diseases showed a very strong downward trend up to 1994, (with a 6-fold decline). Thereafter, a slight increase in deaths for septicaemias, heart infections and hepatitis was recorded. Over time, an increasing proportions of deaths due to infections occurred in the elderly (i. e., > or = 65 years of age), from 48.1% in 1969-1979 to 77.3% in 1990-1999. Mortality rates were consistently higher in men than in women, and showed a substantial geographic heterogeneity. In newborns, from 1969 thru 1999 mortality rates declined 10-fold all over the country, but an inverse north-south geographic gradient persisted during the whole study period. The spread of HIV infection and AIDS epidemic in the first '80s dramatically interrupted the downward trend in infectious disease mortality outlined above. Between 1993 and 1996, HIV/AIDS was the main cause of death among Italian men aged 30 to 39 years. CONCLUSIONS: This statistical analysis allowed to better quantify the impact of infectious diseases on overall mortality in Italy. Observed time trends were in accordance to the picture recorded in other western Countries, whereas the higher newborn mortality in southern Italy reflects the persistence of geographical inequalities in the health care organization. PMID- 15792155 TI - [Agricultural pesticide-related poisonings in Italy: cases reported to the Poison Control Centre of Milan in 2000-2001]. AB - OBJECTIVE: Aim of the present study is to provide a preliminary description of agricultural pesticide-related poisonings in Italy. DESIGN: The study focuses on the cases with at least one sign or symptom due to exposure to agricultural pesticides occurred in 2000-2001 and referred to the Poison control centre of Milan, the Italian centre more frequently consulted nationwide. RESULTS: During the study period, 872 agricultural pesticide-related poisonings were identified. Most of the observed cases occurred in Sicily (n=199), Lombardy (n=100), Apulia (n=99), Veneto (n=75), Emilia-Romagna (n=67) and Campania (n=59). About 86% of the observed cases were unintentional and about 76% of them were men. Children aged 10 or less accounted for about 6% of the total. The poisonings occurred more frequently at home (about 38%) than at workplace (about 24%). The chemical classes of agents more frequently reported were: organophosphates (233 cases), copper and sulphur compounds (140 cases), carbamates (126 cases) and pyrethrins/pyrethroids (102 cases). The top six pesticides associated with poisonings were methomyl (97 cases), dimethoate (59 cases), copper sulfate (58 cases), glyphosate (53 cases), sulphur compounds other than copper sulfate (47 cases), and paraquat dichloride (41 cases). CONCLUSION: The data here presented provide an initial support to identify priorities for prevention in agricultural settings. PMID- 15792156 TI - [Common mental disorders in primary care: prevalence and determinants of a general health questionnaire as a screening tool]. AB - OBJECTIVES: This study intends to evaluate the prevalence of common mental disorders in a population residing in Florence cared for by general practitioners and to assess the determinants of classification and recognition of common mental disorders of the General Health Questionnaire with 12 items as screening tool. (GHQ-12). PARTICIPANTS: The study includes a total of 676 patients received in the periods from June to September 1999 and from January to February 2000 in out patient treatment by 13 general practitioners during a 2 week sampling in the consulting room of each doctor. METHODS: Every patient filled in a tool set composed of socio-demographic form, GHQ-12, Social Readjustment Rating Scale (SRRS). To evaluate the diagnostic reliability of GHQ-12, a subgroup of 158 patients underwent The Mini International Neuropsychiatric Interview (MINI). RESULTS: Among the 676 patients contacted the estimated prevalence (Probable Prevalence) of common mental disorders was of 43.6%. The factors connected to the GHQ-12 high-scorers are gender, number of consultations and life events evaluated through the SRRS. The school level results as protective factor. CONCLUSIONS: This study pointed out the importance of psycho-social factors in determining common mental disorders in the general practice, moreover the prevalence of such disorders is high. PMID- 15792157 TI - [Promoting physical activity: going to school by the Piedibus (walking school bus)]. AB - The Local Health Service of Lecco has proposed the project called Piedibus (Walking school bus), the organisation of safe pedestrian pathways from home to school, in order to promote the increase of physical activity among children. This article describes how the project has been planned and made operational. It provides some results about the community participation and the satisfaction of students, teachers and accompanying volunteers. The project organisation is centered on community participation with large institutional involvement. At the moment (October 2004), independently of weather conditions, 12 Piedibus lines, with more than 500 pupils and 90 accompanying volunteers, are running daily in the Lecco Province. For the next school year 2004-2005 we estimate that the number of lines and users involved will double. The opinion of children, teachers and volunteers has been in general very favourable toward the project: 91.4% of participating students stated they liked the initiative very much and 87.4% among them prefer to go to school by the Piedibus than by any other means. Teacher and volunteers outlined the social value of the project and the increase in physical activity as positive aspects of the project. The overall impact at a province level is outlined by the Media coverage with over 60 articles and numerous radio and television broadcasts. PMID- 15792159 TI - [Case-control evaluation of screening efficacy]. PMID- 15792158 TI - Occupational injuries in Greece (1938-1955): history of medicine and descriptive epidemiology. AB - AIM: To describe the longitudinal trends in the rates of total and fatal occupational accidents in Greece during 1938-1955. MATERIAL AND METHODS: Information on occupational injuries have been provided from the yearly reports of the Organization of Social Insurances (1938-1955) and on population data from the tables of National Statistic Agency. Bio-statistical analysis was performed by the use of SPSS software and Stat-Calc of Epi Info. RESULTS: The evolution of the longitudinal trend of occupational accidents has revealed a biphasic character, with a decreasing trend during 1938-1945 and an increasing trend during 1946-1955. The phenomenon was obvious in both sexes and in all age groups. On the contrary fatal occupational injuries increased across the period 1938-1945 and subsequently decreased. These temporal trends can be interpreted on the light of the important reduction in the level of economic activity during the second world war and the subsequent gradual recovery in the post war period. CONCLUSION: The biphasic characteristics of the occupational accidents longitudinal trend seems to be influenced by historical factors. Important lessons were learnt from the period of war. The decrease of the rate of total occupational accidents does not necessary reflect a satisfactory level of safety at work. The level of the economic activity, the efficiency of the registration and prevention agencies play a role. In addition, the rate of fatal injuries has a critical role in benchmarking national occupational health performance. PMID- 15792160 TI - Magnetic resonance imaging of the infrapatellar fat pad: correlation with patellar articular cartilage abnormalities. AB - We tried to demonstrate an association between magnetic resonance imaging findings of the Hoffa fat pad and patellar chondropathy. Parallely, we checked the correlation between the diagnosis of patellar chondropathy on magnetic resonance imaging and during arthroscopy. Our study is a retrospective review of the records of 135 patients who underwent an arthroscopy and MRI study at our institution between October 1997 and January 2001. Magnetic resonance images of the Hoffa fat pad were interpreted and typewritten arthroscopy reports were recorded. A patellar chondropathy assessed during arthroscopy was present in 64 of 135 patients. Twenty of them had abnormal signal intensity in Hoffa fat pad with a sensitivity of the magnetic resonance imaging findings of 31% and a specificity of 73%. We didn't find any significant association in the different correlations between signal abnormalities of Hoffa fat pad and patellar chondropathy. On the other hand, there was a significant association between the results of patellar chondropathy on magnetic resonance imaging findings and during arthroscopy. No significant association was shown between the MRI findings of Hoffa fat pad and the patellar chondropathy. PMID- 15792161 TI - Post-cardiac catheterization femoral fistula corrected by ultrasound-guided compression. AB - We present a case of post-cardiac catheterization femoral fistula diagnosed by color and pulsed Doppler sonography and treated by ultrasound-guided compression. We avail ourselves of the case to review the echo Doppler semiology of the different types of arteriovenous fistulas with an emphasis on the functional analysis of the dialysis accesses. We also stress the therapeutic options in front of undesirable shunts. PMID- 15792162 TI - Health screening with CT: prospective assessment of radiation dose and associated detriment. AB - The use of computed tomography (CT) for screening of targeted diseases is gaining much interest in the international medical community. An important aspect in the justification of screening with such a high dose examination is the radiation dose to the patient. The objective of this study was to perform a prospective assessment of the radiation dose and associated risk for deleterious effects from a CT screening programme. The excess lifetime risk of fatal cancer for various screening strategies was quantified by estimating the effective dose and by using age dependent mortality risk factors. The accrued data shows that the excess mortality risk due to screening with CT could be substantial. It ranges from 0.01% up to a few percent, strongly depending on the type and method of screening. Consequently, radiation dose and associate risk should be included as fundamental parameters for outlining and deciding a screening approach with CT. Recent technical developments, such as tube current modulation, are promising tools for dose reduction within the constraint of desired image quality. PMID- 15792163 TI - Mri findings in primary non-Hodgkin's lymphoma of the liver. AB - Primary lymphoma of the liver is a very rare malignancy. Most often, these lesions consist of diffuse large B-cell non-Hodgkin's lymphoma that occurs mostly in immunodeficient patients. To prove the primary nature of a hepatic lesion, a systemic lymphoproliferative disease should be ruled out. Secondary liver involvement during Hodgkin's and non-Hodgkin's lymphoma is frequent. In advanced cases the incidence varies from 25% to 50%. The present case describes the MRI features of a primary lymphoma of the liver presenting as a solitary nodule. The primary lymphoma presents as a T2-hyperintense homogeneous nodule, with a signal intensity comparable with the signal intensity of the spleen. Signal intensity is comparable on in and out of phase imaging. The nodule is slightly T1-hypointense and doesn't show any arterial contrast uptake. In the late venous phase a slight increase in signal intensity is noted. Two and a half minutes after the administration of contrast agent, the lesion is iso-attenuating with the liver parenchyma. This case is rare because of the concomitant presence of heterozygous sickle cell anaemia and the presence of Gamna-Gandy bodies in the splenic parenchyma. It remains uncertain whether the presence of the Gamna-Gandy bodies is associated with the liver lymphoma or with the underlying sickle cell anaemia, or with a combination of both. PMID- 15792165 TI - [Radiology versus medical imaging]. AB - In an increasing number of hospitals in Belgium, the radiologists are changing the names from their department from "radiology" to "medical imaging". The reasons behind this change remain unclear and it is certainly not a good move in a time where turf battles will increase. PMID- 15792164 TI - Small bowel obstruction secondary to disseminated candidiasis in an immunocompromised patient: radiologic-pathologic correlation. AB - Immunosuppression for therapeutic reasons (e.g. post transplantation, post chemotherapy), as well as pathologic immunodeficiency due to certain pathologic conditions (e.g. AIDS, leukemia), is increasingly encountered in daily medical practice. As a result, the concomitant risk for opportunistic infections is higher and immunocompromised patients may present with uncommon clinical and radiologic conditions. We report on a case of a 33-year-old immunocompromised woman with a history of recurrent T-cell lymphoblastic lymphoma, which presented with abdominal pain. Computed tomography (CT) images demonstrated significant small bowel dilatation, wall thickening, and high-density intestinal content, with a focal point of transition in the pelvis. Extensive fungal enteritis due to Candida Albicans with partial small bowel obstruction was found on autopsy. PMID- 15792167 TI - Benign bone tumours of the spine. AB - A wide variety of primary bone tumours can involve the spine. The imaging features of these lesions are often characteristic. We present an overview of the primary benign bone tumours of the spine. The role of plain film, CT and MRI is discussed. PMID- 15792166 TI - Ankylosing spondylitis: what remains of the standard radiography anno 2004? AB - In this review, the radiographic features of ankylosing spondylitis of the axial skeleton will be discussed shortly. Three pathologic processes, including inflammation, bony repair and ossification occurring consecutively or simultaneously, will contribute to the radiographic picture of ankylosing spondylitis. Typical target sites at which these processes take place are the synovial joints, discovertebral joints and ligamentous attachments or entheses of the axial skeleton. PMID- 15792168 TI - Congenital abnormalities of the osseous spine: a radiological approach. AB - The spine may act as a useful window to the diagnosis of many congenital malformations syndromes and skeletal dysplasias. However, radiological identification of these syndromes remains a difficult task, because there are so many syndromes and dysplasias to remember. Moreover, many spinal abnormalities are non-specific and there is much overlap between different genetic and congenital disorders. Consequently, many radiologists cringe when these topics are discussed. The purpose of this short review is to provide the general radiologist a workable primer for systematic analysis of spinal abnormalities encountered in genetic disorders, which may be helpful in (differential) diagnosis. PMID- 15792169 TI - Ruptured pulmonary hydatid cyst. PMID- 15792170 TI - Spontaneous remission of an osteoid osteoma of the axis. PMID- 15792171 TI - Balloon valvotomy during pregnancy. PMID- 15792172 TI - What is the role of balloon dilatation for severe aortic stenosis during pregnancy? AB - BACKGROUND AND AIM OF THE STUDY: Severe aortic stenosis in pregnancy creates several challenges for the physician. In recent years, balloon valvuloplasty has become more widely used, though the indications for its use in this setting are unclear. A review of the available evidence is presented, and a suggested management strategy illustrated. METHODS AND RESULTS: Available literature on the subject was reviewed via Medline search and reference lists from the identified articles. Particular attention was paid to prediction of risk, management options and outcome. The data suggest the importance of early symptoms in determining management, as there is a high risk of complications if left untreated. This group should be considered for valvuloplasty, whereas asymptomatic patients are at low risk, and can be managed expectantly. This is illustrated with two contrasting cases from the authors' practice: the symptomatic patient underwent aortic balloon valvuloplasty as a palliative procedure, using transesophageal and minimal fluoroscopic guidance, with good medium-term results. Both patients required aortic valve replacement in the medium to long-term. CONCLUSION: The use of aortic balloon valvuloplasty in pregnancy is useful as a palliative procedure, allowing deferral of valve replacement until after birth. Echocardiographic features alone are not enough to decide on management, and symptoms play a vital role in determining risk. The use of transesophageal echocardiography during the procedure significantly reduces fluoroscopy time. PMID- 15792173 TI - Valve replacement in women of childbearing age: influences on mother, fetus and neonate. AB - BACKGROUND AND AIM OF THE STUDY: Currently, there is no clear consensus on the optimal type of cardiac valve prosthesis that should be placed in women of childbearing age. The risks of reoperative surgery for bioprosthetic valves must be weighed against those of anticoagulation therapy required for mechanical valves. Bioprosthetic valves placed in women of childbearing age are not necessarily superior to mechanical valves in terms of maternal or fetal outcomes. METHODS: A retrospective analysis was performed of early and late outcome in 103 women of childbearing age (mean age 28.3 +/- 5.1 years; range: 18-35 years) who underwent mechanical (n = 63) or biological (n = 40) valve replacement between January 1982 and July 2002. RESULTS: Eleven of the 40 women (28%) with biological valves had 19 pregnancies (13 vaginal births; three caesarean sections). Twelve of the 63 women (19%) with mechanical valves had 37 pregnancies (nine vaginal births, four caesarean sections). All of the women with mechanical valves became pregnant while receiving warfarin, and were subsequently placed on heparin or enoxaparin. No bleeding complications or birth defects were observed in either group. The incidence of miscarriages (32% (12/37) versus 11% (2/19), p = 0.09) and therapeutic abortions (32.4% (12/37) versus 5.3% (1/19), p = 0.06) were greater in the group with mechanical valves than with bioprosthetic valves. Freedom from reoperation at five and 10 years for biological versus mechanical valves was 79% versus 90%, and 38% versus 82%, respectively (p < 0.01), with no reoperative mortality. CONCLUSION: Biological valves are recommended in women of childbearing age, despite the fact that no birth defects were seen in children born to women with mechanical valves on warfarin, because the teratogenic effects of warfarin may be masked by the high rate of miscarriages and therapeutic abortions in this group. PMID- 15792174 TI - Pediatric mitral valve replacement: incremental risk factors impacting survival and reintervention. AB - BACKGROUND AND AIM OF THE STUDY: Predictors of survival and reintervention in children undergoing mitral valve replacement (MVR) were assessed. METHODS: A total of 37 children underwent MVR, with a mean prosthesis size of 22.5 mm. RESULTS: Six patients died during the study. Actuarial and event-free survivals (including operative mortality) at 10 and 20 years were 85 and 85%, and 55 and 37%, respectively. The linearized rate of bleeding was 0.67% per patient-year (pt yr). There were no instances of thromboembolism, endocarditis or structural valve dysfunction. Six patients required reintervention for somatic outgrowth at a mean postoperative interval of 104 months. Freedom from reintervention at five, 10 and 20 years was 92, 73 and 48%, respectively. There were no deaths or complications following reintervention. The mean follow up was 96 months (range: 3 to 276 months); total cumulative follow up was 296 pt-yr. Multivariate predictors of mortality were age < 6 months, NYHA class IV, congestive cardiac failure, internal orifice diameter (IOD) < 14 mm and effective orifice area (EOA) < 1.5 cm2. Predictors for reintervention were age < 6 months (p = 0.0021), predicted annulus size < 16 mm (p < 0.0001), IOD < 18 mm (p = 0.05), IOD index (IODI) > 50 mm/m2 (p = 0.007), EOA < 2.5 cm2 (p = 0.05), EOA index (EOAI) > 7 cm2/m2 (p < 0.0001) and prosthesis size-body surface area (BSA) index (SI) > 69 mm/m2 (p = 0.007). CONCLUSION: Early mortality occurs in neonates and infants with associated lesions, particularly atrioventricular septal defect. Risk factors for mortality include age, NYHA class, and small IOD and EOA. Risk factors for reintervention include age < 6 months, small predicted annulus size, IOD and EOA, IODI > 50 mm/m2, EOAI > 7 cm2/m2 and SI > 69 mm/m2. PMID- 15792175 TI - Mechanical aortic valve replacement: long-term outcomes in children. AB - BACKGROUND AND AIM OF THE STUDY: Early and late outcomes following mechanical aortic valve replacement (AVR) in children were analyzed. METHODS: Between January 1980 and December 2003, 55 patients underwent mechanical AVR at the authors' institution. Of these patients, 12 had aortic regurgitation (AR), 31 had aortic stenosis (AS), and 12 had mixed disease. Preoperatively, 25 patients (45.5%) were in NYHA classes III and IV. Among patients, 22 had a previous valvotomy and 19 had associated cardiac defects. Isolated AVR was performed in 37 cases. Twelve patients required root enlargement procedures, two had aortic root replacement, three had a double valve replacement, and one patient had a concurrent mitral annuloplasty. The mean prosthesis size was 22.6 mm (range: 16 31 mm). Mean follow up was 12.28 +/- 6 years (range: 1-23 years); total follow up was 665 patient-years (pt-yr). RESULTS: There was one late death. Actuarial survival at 20 years was 98 +/- 2%. Event-free survival at one, five and 20 years was 96 +/- 3%, 92 +/- 4% and 88 +/- 5%, respectively. Four patients required reoperation (two for valve outgrowth, one each for paravalvular leak and prosthetic valve endocarditis (PVE)). Freedom from reintervention at one, five and 20 years was 98 +/- 2%, 96 +/- 3% and 92 +/- 4%, respectively. There was one event of anticoagulation-related hemorrhage. Freedom from anticoagulant-related hemorrhage at 20 years was 98 +/- 2%, and freedom from PVE at five and 20 years was 98 +/- 2% and 96 +/- 3%, respectively. There were no instances of thromboembolism or structural valve dysfunction. Linearized rates of bleeding and endocarditis were 0.15 and 0.3% per pt-yr, respectively. At follow up, 54 children were in NYHA classes I or II. CONCLUSION: Mechanical AVR, with aortic root enlargement if necessary, is associated with low mortality and morbidity, and is an excellent treatment option in children. Late embolic and hemorrhagic complications are infrequent in the current era. PMID- 15792176 TI - Seventeen-year clinical results of 1,037 Mitroflow pericardial heart valve prostheses in the aortic position. AB - BACKGROUND AND AIM OF THE STUDY: It is generally believed that the pericardial tissue valve is associated with a greater propensity to early structural failure and subsequent valve-related complications. However, a modified valve design during recent years has considerably improved the clinical durability of this bioprosthesis. METHODS: Between April 1986 and December 2003, single Mitroflow pericardial valve prostheses were inserted in the aortic position in 1,037 patients (mean age 72.4 +/- 8.4 years). Coronary artery bypass grafting (CABG) was performed in 373 patients (36%). Preoperative echocardiographic findings of structural valve deterioration (SVD) were confirmed at reoperation. Total follow up was 5,022 patient-years (pt-yr) (mean 4.85 +/- 4.7 years) and was 99.7% complete (three patients lost). RESULTS: Early (30-day) mortality was 2.9% for elective surgery; 434 patients (41.9%) died during the follow up period. Actuarial survival at 17 years was 23.2 +/- 2%. Actuarial freedom from valve related mortality at 17 years was 92.0 +/- 2.2%, whereas the linearized mortality rate was 0.4%/pt-yr. Linearized valve-related complications occurred at a rate of 3.6%/pt-yr (n = 122). SVD of Mitroflow bioprostheses occurred at a rate of 1.0%/pt-yr, and caused stenosis (36.7%), insufficiency (20.4%) and combined stenosis and insufficiency (42.9%). Actual freedom from SVD at 17 years was 95.3 +/- 0.7% for all patients, 96.2 +/- 0.6% for those aged > 60 years, and 99.0 +/- 0.4% for those aged > 70 years. CONCLUSION: The Mitroflow pericardial bioprosthesis, in the aortic position, provides an excellent clinical performance and a low rate of valve-related morbidity, mortality and reoperation for up to 17 years. The prosthesis also demonstrates satisfactory hemodynamic performance, and is therefore recommended for patients aged > or = 70 years and for those aged 60 69 years with comorbidities. PMID- 15792177 TI - Reduction of the QT interval dispersion after aortic valve replacement reflects changes in electrical function rather than structural remodeling. AB - BACKGROUND AND AIM OF THE STUDY: Factors related to changes of QT dispersion (QTd) after aortic valve replacement (AVR) in patients with aortic stenosis were analyzed. METHODS: The prospective group comprised 121 consecutive patients (45 women, 76 men; mean age 58 +/- 11 years; range: 24-77 years) with significant aortic valve stenosis. Data (clinical, echocardiographic and electrocardiographic) were collected before and at least 16 months after AVR. QTd was measured in the standard ECG. RESULTS: Before AVR, the mean QTd was 60 +/- 24 ms (QT(max) 424 +/- 40 ms). QTd was > 50 ms in 68% of patients, and > 70 ms in 30%. During postoperative follow up the mean QTd was 54 +/- 19 ms (QT(max) 368 +/ 36 ms) for all patients, and was > 50 ms in 58% of cases and > 70 ms in 13%. Postoperatively, QTd was decreased to < 70 ms in 27% of patients with a normalized left ventricular mass index (LVMI), and in 27% of those without any clinically significant reduction in left ventricular (LV) hypertrophy. In the multivariate analysis, QTd reduction was weakly related to the reduction in LV wall thickness (p = 0.09) and LVMI (p = 0.05). The reduction in QTd was more related to changes in T-wave amplitude in lead V5 (p = 0.004). CONCLUSION: Following AVR for aortic stenosis, a decrease in QTd was observed, notably among patients with QTd > 70 ms. This reduction was only weakly related to the degree of reduction in cardiac hypertrophy, but a more important relationship was observed with changes in T-wave amplitude. These findings suggest that a reduction in QTd after AVR is reflective of changes in electrical function rather than structural remodeling. PMID- 15792178 TI - Acoustic diagnosis of aortic stenosis. AB - BACKGROUND AND AIM OF THE STUDY: Phonocardiography is a promising non-invasive diagnostic tool for the assessment of aortic stenosis (AS), and time-frequency representation is a potential tool to extract information from the phonocardiogram (PCG) signal. The study aim was to develop an acoustical method to predict the severity of AS. METHODS: Normalized continuous wavelet transform (NCWT) and fast Fourier Transform (FFT) were used to perform a spectral analysis of the PCG signal. A multi-peak detection algorithm was developed to determine the dominant frequency (DF) of systolic murmurs (SM). The spectral ratio of the SM, integration of the NCWT of SM (SI), and combined information of SM and second heart sound, were also calculated. RESULTS: The DF correlated best with the hemodynamic data: r = -0.72 with aortic valve (AV) area; r = 0.63 with maximal blood velocity through the AV; and r = 0.57 with mean pressure gradient across the AV. Based on DF and SI data, the study subjects (n = 59) were classified into three categories: severe AS; moderate AS; and other cases. The acoustical and echo classifications were in agreement in 50 subjects (85%). CONCLUSION: The acoustical method developed cannot predict accurately the severity of AS, but is valuable when conducting a screening classification before an invasive method is used. PMID- 15792179 TI - Anterior mitral basal 'stay' chords are essential for left ventricular geometry and function. AB - BACKGROUND AND AIM OF THE STUDY: Among the anterior mitral basal chords, two particularly strong and thick stay chords (SC) remain under tension during the entire cardiac cycle. Collagen fibers of the anterior mitral leaflet (AML) are oriented from insertion of the SC on the AML to the fibrous trigones (FT), suggesting that local stress is directed from the papillary muscles (PM) over the SC and AML to the FT, maintaining left ventricular (LV) geometry. METHODS: Sonomicrometry crystals were implanted in sheep at the LV apex (A), the anterior (AW) and septal (SW) LV wall, the PM tips (M1 and M2), the SC insertion into the AML (S1 and S2), the posterior (PMA) and lateral (P1 and P2) mitral annulus, the FT (T1 and T2), the tips of the anterior (AL) and posterior (PL) mitral leaflets, and the base of the aortic right coronary sinus (RCS). Changes in distances, areas, and volume were time-related to aortic flow and LV and ascending aorta pressures. Recordings were taken at baseline and after transection of the SC. RESULTS: After transection of the SC, the systolic distance from M1-T1 increased by +0.96 +/- 0.41 mm (p < 0.05) and from M2-T2 by +0.97 +/- 0.42 mm (p < 0.05). The LV length increased at T1-A by +1.14 +/- 0.60 mm (p < 0.05) and at T2-A by +0.97 +/- 0.37 mm (p < 0.05). The aortomitral angle narrowed at end-systole by 3.26 +/- 0.85 degrees (p < 0.05). Transection of the SC reduced dP/dt by -11.20 +/- 5.29% (p < 0.05), maximum aortic flow by -16.89 +/- 7.86% (p < 0.05), and maximum pressure-volume ratio by -10.83 +/- 3.36% (p < 0.05). CONCLUSION: Transection of the anterior mitral SC did not result in mitral regurgitation but induced significant changes in LV geometry, including narrowing of the aortomitral angle and subsequent deterioration of LV function. The SC are essential for maintaining normal LV geometry and function. PMID- 15792180 TI - Influence of different echocardiographic imaging modes on the assessment of anterior mitral leaflet thickness. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to compare fundamental imaging (FI) measurements with harmonic imaging (HI) measurements and surgical measurements (SM) in the assessment of mitral anterior leaflet thickness. METHODS: Forty-three patients scheduled to undergo mitral valve replacement were included. Before surgery, routine echocardiography was performed in all patients using an instrument fitted with a 2.5 MHz broadband transducer. The anterior mitral leaflet was measured with different echocardiographic imaging modalities, and also surgically. During FI, the transducer transmission frequencies were 2.5 MHz and 2 MHz, while transmission frequencies of 1.7 MHz and 1.5 MHz were used during HI. Surgical measurements were taken immediately after surgery from a mitral valve specimen that was removed intact. RESULTS: Mitral anterior leaflet thickness measurements derived from FI at 2 MHz transmission frequency (3.8 +/- 1.1 mm, p = 0.020), HI at 1.7 MHz (4.4 +/- 1.2 mm, p < 0.001), and HI at 1.5 MHz (4.5 +/- 1.2 mm, p < 0.001) were significantly larger than those made surgically (3.3 +/- 0.6 mm). However, no significant differences were seen between thickness measurements derived from FI at 2.5 MHz transmission frequency and SM (3.7 +/- 1 mm versus 3.3 +/- 0.6 mm, p = 0.063). Mitral anterior leaflet thickness was greater with HI than with FI (1.7 MHz versus 2.5 MHz, p < 0.001; 1.7 MHz versus 2 MHz, p < 0.005; 1.5 MHz versus 2.5 MHz, p < 0.001; 1.5 MHz versus 2 MHz; p < 0.002). The closest measurement to SM was that obtained at a transmission frequency of 2.5 MHz (mean difference 0.3 +/- 0.6 mm; SEE 0.1 mm). CONCLUSION: Mitral anterior leaflet thickness measurements made with HI appear to be greater than those made with either FI or SM. PMID- 15792181 TI - A valve replacement technique for heavily calcified mitral valve and annulus. AB - Severe mitral annular calcification is frequently encountered in patients with chronic renal failure requiring dialysis. The procedures are described for the implantation of mitral prostheses in two dialysis patients with mitral stenosis who exhibited heavily calcified mitral annuli. In both cases the calcifications extended around the entire mitral annulus as well as the valve leaflets, papillary muscle, atrial wall and ventricular myocardium, making it impossible to secure prosthetic valves to the mitral annuli. Before implantation, the calcification was partially debrided ultrasonically, and an 'atrioventricular channel' created. A mechanical valve with a polyester fabric collar was fixed on the atrial side of the calcified annulus using double sutures and gelatin resorcin-formol glue. Perivalvular leakage, obstruction of valve opening or tortuous movements of prosthetic valve ring were not observed, and neither patient developed heart failure after surgery. This technique represents a potentially useful approach in the surgical treatment of patients with severely calcified mitral annuli. PMID- 15792182 TI - Decellularization of bovine pericardium for tissue-engineering by targeted removal of xenoantigens. AB - BACKGROUND AND AIM OF STUDY: Assessment of decellularization of xenogeneic biological scaffolds for tissue engineering has relied primarily on histological cellularity, though this may not ensure the removal of known xenogeneic antigens such as galactose-alpha1,3-galactose (alpha-gal) and MHC I. METHODS: Bovine pericardium (BP) underwent standard (Std) decellularization consisting of hypotonic lysis and treatment with DNAse/RNAse. In addition to Std decellularization, tissues were treated for 24 h with either 0.5% Triton X-100, 0.5% sodium deoxycholate (SD), 0.1% sodium dodecyl sulfate (SDS), alpha galactosidase (5 U/ml) or phospholipase (PL) A2 (150 U/ml). Tissues underwent a 96-h washout under gentle agitation at 27 degrees C, and then evaluated by light microscopy for % cellularity, and by immunohistochemistry and Western blot for alpha-gal, bovine MHC I and smooth muscle alpha-actin. RESULTS: Standard treatment of BP resulted in only partial removal histological cellularity and persistence of alpha-gal, MHC I and alpha-actin. Adding SD treatment resulted in apparent acellularity, but persistence of xenogeneic antigens. Only the addition of SDS resulted in complete histological acellularity and removal of xenogeneic antigens. Treatment with alpha-galactosidase selectively removed alpha-gal from BP. CONCLUSION: Histological cellularity is not an adequate end-point for assuring removal of antigenicity from xenogeneic biological scaffolds. However, known xenogeneic antigens can be targeted for removal by novel decellularization treatments such as alpha-galactosidase. PMID- 15792183 TI - Localization and pattern of expression of extracellular matrix components in human heart valves. AB - BACKGROUND AND AIM OF THE STUDY: The pattern of expression and distribution of extracellular matrix (ECM) components in human cardiac leaflets was analyzed. Additionally, interstitial cells (ICs) from the four different leaflets were isolated and studied. METHODS: Immunohistochemistry and immunocytochemistry were used for localization, and flow cytometric analysis to quantify the expression of specific markers on these ICs; the synthesis and expression of ECM components was assessed. RESULTS: Elastin was found predominantly on the inflow layer, but fine fibers were also present in the central and outflow layers. Collagen I was predominantly on the outflow layer but permeated throughout the leaflets. Collagen III was expressed ubiquitously. Proteoglycan expression was throughout the leaflet, but was predominant in the central layer. Fibronectin and vitronectin were expressed strongly in the inflow layer, moderately in the central layer, and weakly in the outflow layer. Biglycan expression was ubiquitous, with strong filamentous strands in the central layer. Keratan sulfate and decorin were ubiquitous. Chondroitin-4-sulfate and chondroitin-6-sulfate were strongly expressed in the outer layers, and laminin was restricted to the basal lamina of the endothelial cells. Cultured ICs showed synthesis and expression of various ECM components. CONCLUSION: This study of the pattern of expression of ECM components may provide a basis for a fingerprint on which to base future valve alternatives. The results provide useful information for valve tissue engineering and an understanding of the structural basis of some sophisticated functions of the valves. PMID- 15792184 TI - Biocompatibility and recellularization potential of an acellular porcine heart valve matrix. AB - BACKGROUND AND AIM OF THE STUDY: Tissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. The study aim was to investigate the biocompatibility and recellularization potential of an acellular porcine valve matrix. METHODS: Acellular porcine valve matrix contact and extract cytotoxicity was tested against porcine fibroblasts and smooth muscle cells (SMC). Porcine cells were incubated with decellularized aortic valve leaflets and aortic wall, and then assessed for changes in morphology and contact inhibition of growth. Soluble tissue extracts were prepared from decellularized leaflets and aortic wall, and assessed for their effect on the viability of cultured porcine cells. Acellular leaflets were seeded with either fibroblasts or SMC at 1 x 10(3) to 1 x 10(6) cells/cm2 for 24 h, or 5 x 10(4) cells/cm2 for 1-4 weeks. Cell attachment onto, and migration into, the acellular matrix was assessed by scanning electron microscopy and histology. RESULTS: No contact inhibition of growth, or changes in fibroblast or SMC morphology, were observed following contact with the acellular valve matrix. No soluble extract cytotoxicity was found. Intermediate cell-seeding densities (2.5 x 10(4) to 7.5 x 10(4) cells/cm2) of both cell types produced confluent cell attachment; at the lowest concentration (1 x 10(3) cells/cm2) cell attachment was sparse, and at the highest (1 x 10(6) cells/cm2) it was multilayered. The SMC migrated throughout the leaflet matrix over four weeks, but there was no fibroblast migration into the matrix. CONCLUSION: The absence of contact and extract cytotoxicity indicated that the acellular valve matrix was biocompatible in vitro. The failure of porcine fibroblasts to grow on, or infiltrate into, the matrix suggested that the SMC may be the preferred cell type for future leaflet recellularization studies in the development of a tissue-engineered heart valve replacement. PMID- 15792185 TI - Primary valvular surgery in octogenarians: perioperative outcome. AB - BACKGROUND AND AIM OF THE STUDY: Cardiac surgery in octogenarians is now performed routinely, and generally results in a good improvement in functional capacity. The study aim was to evaluate operative mortality and to identify preoperative and postoperative risk factors of mortality. METHODS: A total of 200 consecutive patients (79 males, 121 females; mean age 83 years; range: 80-90 years) who underwent valvular surgery at the authors' institution between 1991 and 2002 was reviewed. Among patients, 154 underwent aortic valve replacement (77.0%), 35 mitral surgery (17.5%), and 11 aortic and mitral valve surgery (5.5%). Forty-five patients (22.5%) had concomitant myocardial revascularization, and 23 (11.5%) were operated on in an emergency setting. Preoperative risk factors studied included endocarditis (2.0%), ventricular dysfunction (22.5%), pulmonary hypertension (33.5%), renal dysfunction (31.5%), chronic obstructive pulmonary disease (7.0%) and arteriopathy (9.0%). The mean EuroSCORE, which was used to assess predicted operative risk, was 9.1; the score was < 9 in 104 patients (52%) and > 9 in 96 (48%). RESULTS: Hospital mortality was 7% (n = 14). There were no significant preoperative risk factors of mortality. Postoperative complications occurred in 115 patients (57.5%), including low cardiac output (16.0%), supraventricular arrhythmia (29.5%), pulmonary complications (9.5%), gastrointestinal ischemia (2.0%), wound infection (2.0%) and surgical re exploration (5.5%). Low cardiac output (p < 0.001), gastrointestinal ischemia (p = 0.03) and surgical reexploration (p = 0.004) were significant risk factors of mortality. CONCLUSION: Valvular surgery in octogenarians is a safe and low-risk procedure. The present data tended to show that the EuroSCORE overestimates mortality in this group of patients. PMID- 15792186 TI - Does EuroSCORE predict length of stay and specific postoperative complications after heart valve surgery? AB - BACKGROUND AND AIM OF THE STUDY: EuroSCORE is the most rigorously evaluated scoring system in cardiac surgery. The study aim was to evaluate the performance of EuroSCORE in the prediction of in-hospital postoperative length of stay and specific major postoperative complications after heart valve surgery. METHODS: Data obtained from 1,105 consecutive patients who underwent isolated or combined heart valve surgery were collected prospectively. The EuroSCORE model (standard and logistic) was used to predict in-hospital mortality, prolonged length of stay (> 20 days) and major postoperative complications. A C statistic (receiver operating characteristic curve) was used to test discrimination of the EuroSCORE. Calibration of the model was assessed by the Hosmer-Lemeshow goodness-of-fit statistic. RESULTS: In-hospital mortality was 6.3%, and 21.7% of patients had one or more major complication. EuroSCORE showed very good discriminatory ability in predicting postoperative renal failure (C statistic: 0.78) and good discriminatory ability in predicting in-hospital mortality (C statistic: 0.72), prolonged length of stay (C statistic: 0.71), stroke over 24 h (C statistic: 0.73), gastrointestinal complications (C statistic: 0.73) and respiratory failure (C statistic: 0.71). There were no differences in terms of the discriminatory ability between standard and logistic EuroSCORE. The standard EuroSCORE model showed good calibration in predicting these outcomes (Hosmer-Lemeshow: p > 0.05). The logistic EuroSCORE model showed good calibration, except for prolonged length of stay and respiratory failure. CONCLUSION: EuroSCORE can be used to predict not only in-hospital mortality, for which it was originally designed, but also prolonged length of stay and specific postoperative complications such as renal failure, stroke over 24 h, gastrointestinal complications and respiratory failure within the whole context of heart valve surgery. These outcomes can be predicted accurately using the standard EuroSCORE, which is very easily calculated. PMID- 15792187 TI - Contribution of selected serum inflammatory mediators to the progression of chronic rheumatic valve disease, subsequent valve calcification and NYHA functional class. AB - BACKGROUND AND AIM OF THE STUDY: The mechanism of the underlying principle of the progression of chronic rheumatic valve disease (RVD) and subsequent valve calcification are yet not clearly understood. The study aim was to determine whether serum markers of inflammation impact on the severity of chronic RVD, subsequent valve calcification and NYHA functional class. METHODS: The study group comprised 92 patients (27 males, 65 females; mean age 40 +/- 14 years) with RVD; the control group included 50 age- and gender-matched subjects without echocardiographic signs of RVD. All patients underwent echocardiographic of rheumatic valve severity, valve calcification and NYHA functional class. Levels of cytokines (interleukin-6 (IL-6), interleukin-2 receptor (IL-2R), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha)) and serum inflammatory markers (fibrinogen, high-sensitive C-reactive protein (hs-CRP)) were measured in all subjects. RESULTS: Plasma levels of IL-6, IL-8, IL-2R, TNF-alpha and hs-CRP were significantly higher in patients with RVD than in controls (p < 0.001). Significant correlations were identified between mitral score and fibrinogen (p = 0.002), IL-6 (p = 0.007), TNF-alpha (p < 0.001) and hs-CRP levels (p < 0.001). Fibrinogen, hs-CRP, IL-6, TNF-alpha and IL-2R levels correlated with functional class severity, while IL-6 and TNF-a levels correlated strongly with valve calcification (p < 0.001). CONCLUSION: The chronic phase of RVD is associated with ongoing serum inflammatory mediators which correlate strongly with the severity of valve involvement, valve scarring, subsequent valve calcification and decreasing functional status. Future research in this area should focus on whether anti-inflammatory drugs might reduce progression, morbidity and mortality in patients with chronic RVD. PMID- 15792188 TI - Geometrical model of the pulmonary root. AB - BACKGROUND AND AIM OF THE STUDY: The renaissance of the Ross procedure has attracted much attention to the clinical anatomy of the pulmonary valve. Within the reviewed literature which emphasizes the morphology of the pulmonary root, no detailed anatomy has been reported. The study aim was to describe the cardiosurgical-orientated anatomy of the pulmonary valve. METHODS: Morphometric measurements were made on 20 fixed normal pulmonary roots. The roots were fixed under diastolic pressure (5 mmHg) with closed leaflets. The distances between the commissures, and between the intervalvular triangles, were measured. The heights of the sinuses and of the intervalvular triangles were also defined. RESULTS: The mean (+/- SD) of each parameter was calculated. The distances of the right, left and anterior sinus commissures were approximately similar, as were the distances of the intervalvular triangles at the root base. The mean height of the left coronary sinus was maximal, followed by the right and anterior pulmonary sinuses. The left intervalvular triangle was highest, followed by the posterior and right intervalvular triangles. The difference between the height of the left sinus and right intervalvular triangle indicated a tilt angle of 16.26 degrees between the root base and sinotubular junction. CONCLUSION: Based on these measured parameters, a spatial geometrical model was constructed which clearly described the asymmetrical structure of the pulmonary root. PMID- 15792189 TI - Replacement of mechanical mitral valve prosthesis due to patient intolerance of clicking noise: case report. AB - A 55-year-old male underwent combined coronary artery bypass grafting and mitral valve replacement with a No. 31 St. Jude Medical mechanical prosthetic valve. Over the next few months, the patient experienced severe difficulty with the 'clicking' noise of the mechanical valve, to the extent that it affected his daily activities. At approximately four months after the first operation, and at the patient's insistence, he underwent redo sternotomy and replacement of the mechanical mitral valve with a Mosaic porcine bio-prosthesis, with an uneventful recovery. While this case represents an unusual indication for valve replacement, it illustrates the importance of considering non-traditional factors when choosing the optimal prosthetic valve. PMID- 15792190 TI - Tricuspid valve repair for biopsy-induced regurgitation in a heart transplant recipient. AB - Tricuspid regurgitation (TR) is a frequent complication after heart transplantation. The etiology of TR is multifactorial, but biopsy-induced flail leaflet is one of the most important mechanisms. A 61-year-old woman underwent heart transplant, but experienced several rejection episodes which required multiple surveillance endomyocardial biopsies. At three months after transplant, she required tricuspid valve repair due to symptomatic severe TR. The anterior leaflet was flail, with rupture of primary and secondary chordae. Valve repair was performed with a triple leaflet edge-to-edge technique. The procedure consisted of suture fixation of the prolapsed anterior leaflet joining to the septal and posterior leaflets, and placement of a 30-mm annuloplasty ring. The patient was uneventfully discharged home on day 7 with trivial TR. At a four years post-transplant evaluation, she was in NYHA functional class I, with preserved ventricular function and trivial TR. She has been followed closely because of post-transplant coronary artery disease. PMID- 15792191 TI - Mitral valve replacement in a patient with severe thrombophilic status and direct antiglobulin test positivity: case report. AB - The case is reported of a patient with a symptomatic thrombophilic status and a positivity for both direct and indirect antiglobulin tests who underwent mitral valve replacement. Because of the antiglobulin test positivity, the patient was enrolled in a preoperative autologous blood donation program in order to avoid exposure to allogenic blood. The operation was successful, and the postoperative course uneventful; the patient was discharged on postoperative day 6 and prescribed oral anticoagulation. This case suggests that a safe valve replacement can be carried out even in the presence of a symptomatic thrombophilic preoperative status. PMID- 15792192 TI - Fatal Burkholderia cepacia early prosthetic valve endocarditis: a very rare case and a review of the literature. AB - A 58-year-old woman who underwent mitral valve replacement and tricuspid valve repair developed severe acute systemic illness four weeks after surgery. Serial blood cultures grew Burkholderia cepacia at four to five days after incubation. Transthoracic echocardiography had confirmed a diagnosis of prosthetic valve endocarditis. The patient did not respond to empirical or culture-sensitive antibiotic therapy of endocarditis. Trimethoprim-sulfamethoxazole (TMP-SMX) therapy was only commenced at three weeks after the start of fever because of delayed presentation. Every effort should be made to identify this organism in the laboratory. When clinically indicated, early presumptive antibiotic therapy with TMP-SMX should be started. PMID- 15792193 TI - In response to: Chambers J, Rimington H, Rajani R, Hodson F, Blauth C. Hemodynamic performance on exercise: comparison of a stentless and stented biological aortic valve replacement. J Heart Valve Dis 2004;13:729-733. PMID- 15792194 TI - A novel device for the non-invasive measurement of free hemoglobin in blood bags. AB - The production of red blood cell concentrates from human donors is a very expensive procedure and human resources are in short supply. Under perfect storage conditions at a temperature of 2-6 degrees C, a blood bag must be used within 35-49 days (in Germany). Visual inspection of the bag for apparent hemolysis by a blood bank physician is a crucial but subjective quality control assessment. Since an interruption of the cold chain cannot be definitely ruled out, bags are often disposed of prematurely for safety reasons. There is currently no method of testing a closed blood bag with respect to hemolysis for its suitability to be used in a transfusion. The proposed optical measuring device is a hemoglobin sensor which determines the free hemoglobin in standard erythrocyte concentrates without opening the bag. The optical measurements are done on the flexible tube connected to the main bag. The optical measurements were evaluated using standard hemoglobin solutions with an accuracy of 0.005 g/dL. These investigations show that in the future each blood bag can be tested non-invasively for its content of free hemoglobin. This will contribute to decreasing the wastage rate of red blood cell concentrates. PMID- 15792195 TI - [Methods of resolution for haptic assistance during catheterization]. AB - During catheterization navigation within the patient is mainly dependent on a live x-ray image on the screen. Although methods for 3D visualisation and remote navigation of the catheter are discussed and tested still precise positioning is merely the result of intense training and a high skill and level of training of the performing surgeon. This article refers to a system which can be considered as an add-on for existing procedures of catheterization. It compromises of a miniaturised force sensor located at the tip of guide-wires whose prototype is shown here. The measured forces will be presented to the surgeon amplified by an external actuator described in this article. As a result a haptic perception of the forces between the tip of the guide-wire and the vessels walls will be available and enable the surgeon to gain an impression which is comparable to palpation of living vessels from the inside PMID- 15792196 TI - Cutaneous microcirculation is not altered by a weak 50 Hz magnetic field. AB - Electromagnetic hypersensitivity (EH) is an increasing problem in modern industrial societies. As crawling sensations are frequently mentioned by EH patients alterations in cutaneous microcirculation possibly linked to exposure to magnetic fields might be involved in the development of such sensations and further dysesthesias. In seven healthy volunteers and in three persons convinced to suffer from EH the microcirculation of the right thumb was determined by laser Doppler-flowmetry (LDF) during exposure to circularly polarized 50 Hz magnetic flux densities of 96 mT. During field exposure the LDF values remained constant. The LDF ratio "field on/field off" was found to be 1.03 +/- 0.03. In contrast, reactive hyperemia and hyperventilation caused significant changes in the LDF values of volunteers as well as of EH patients. Following arterial congestion of the forearm microcirculation of the thumb was clearly increased during reperfusion, and the LDF values were elevated up to 2.02 +/- 0.36. 10 deep breaths caused a significant decrease in the LDF values up to 0.63 +/- 0.18. In conclusion, reactive hyperemia and hyperventilation caused clear alterations of cutaneous microcirculation, whereas, 50 Hz magnetic fields had no influence on cutaneous microcirculation. PMID- 15792197 TI - [Semiautomatic procedure for the investigation of synchronized activity of EEG and heart rate--examination of preterm births]. AB - Recordings of the electroencephalogram (EEG) and of the heart rate variability (HRV) of preterm neonates can give important information on the actual state of the nervous system. Both signals, EEG and HRV, are affected by parameters such as gestational age, stage of maturation and behavioral state. This work describes a method for automatic detection of slow wave EEG-bursts and a tool to average changes in the EEG and the corresponding heart rate. The detection is based on the hjorth activity (HA), calculated from the EEG. HA spikes (HAS) are identified by the determination of the beginning and end of existing spikes. HAS maxima and the time between two consecutive HAS are the basis for the triggering of the bursts. EEG power and time synchronized HR changes are averaged with a time window length of 20 s. Resultant, HR increase and duration are determined. These parameters, obtained by the automatic detection, proved to be comparable to the results of an expert. PMID- 15792198 TI - Characterization of mode II-wear particles and cytokine response in a human macrophage-like cell culture. AB - Informations about wear particles in metallosis (mode II wear) and their effects in vitro and in vivo are limited. The aim of this study was to characterize wear particles obtained intraoperatively and to analyse their effects on cytokine response in an established human macrophage-like cell culture model. METHOD: Wear particles were obtained intraoperatively from four patients with metallosis resulting from CrCoMo/PE/TiAIV-implants (mode II wear) (3 knee, 1 hip prosthesis). After purification, particles were characterized regarding to their composition and size (particle size analyser, electron microscopy, edx-analysis, histological slices). The effects of particles on the release of cytokines (PDGF, IL-1beta, IL-8, TNF alpha) were determined in an established human macrophage like cell culture system by ELISA-assays. RESULTS: The metal wear particles consisted of TiAIV with a mean size of 0.1 +/- 0.15 microm, independent of the prosthesis location. CrCoMo particles could not be detected. In the cell culture model 1456 x 10(8) particles per 1 x 10(6) macrophages released maximum amounts of TNFalpha (8-fold) and IL-8 and IL-1beta (5-fold) while the survival rate of the cells was more than 90 percent. A particle-dependent increase of PDGF-levels could not be detected. CONCLUSION: As already shown for mode I wear particles (contact between primary bearing surfaces), also mode II wear particles cause release of bone resorbing cytokines in a macrophage-like cell culture model. Because their local and systemic effects in vivo are still not completely understood, we recommend a complete removal of wear particles in cases of metallosis to avoid possible immunological reactions of the body as well as periprosthetic osteolysis. PMID- 15792200 TI - Eponyms: errors in clinical practice and scientific writing. AB - Eponymous terms are in daily use in medicine. This system of nomenclature which simply commemorates a person is inconvenient, poses difficulties to students and leads to frequent mistakes in scientific writings. Nevertheless it can be helpful in completely describing a multi-symptomatic medical condition or a complex surgical procedure which otherwise would not be neatly encapsulated in a reasonably convenient word or phrase. We used Finkelstein's test as an example to demonstrate that the use of such nomenclature in clinical practice and scientific writing creates inaccuracies. We contacted 62 consultant orthopaedic surgeons and 47 specialist orthopaedic registrars of whom 53 consultant and 39 registrars responded. Three different descriptions of Finkelstein's test were used as described in current literature. Only 10 (10.7%) surgeons recognised the correct method as described by Finkelstein and 83 (89.3%) were unable to do so. The results shows that a statistically significant proportion of surgeons uses the test (p < 0.0001) but fails to identify the correct method (p < 0.0001). We also found that Finkelstein's test was inaccurately described in literature since Leao's incorrect description in 1958 (quoting Eichhoff's manoeuvre) and the mistake persisted for over 50 years before it could be accredited. Such mistakes are frequent not only in hand surgery but in other sections of medicine as well. We conclude that in the modern era of evidence based medicine, use of such trivial nomenclature should be avoided. Efforts should be made to introduce proper descriptive nomenclature, by devising criteria systems which would be easy to use and not fictitious. PMID- 15792199 TI - Cementless Vektor-titan stem in total hip arthroplasty. AB - INTRODUCTION: Cementless THR is a well established, and a widely accepted optimal procedure for younger patients. The cementless Vektor-Titan stem is made of Ti6AI7Nb, has got the shape of a three-dimensional cone, and an optimal proximal anchoring property. MATERIALS AND METHODS: The aim of this prospective study was to scrutinise the outcome of 250 Vektor-Titan stems in cementless THRs with an average follow-up time of 3.0 years (Min: 1, Max: 6). The average age of the patients including 148 women and 102 men was calculated with 54.6 years (Min: 22.5, Max: 77.7). RESULTS: The score according to Merle d'Aubigne improved from preoperative 9.3 (Min: 7, Max: 13) to postoperative 17.0 (Min: 14, Max: 18). Distal cortical hypertrophy and proximal atrophy was detected in 4 cases. Single atrophy of the proximal femur was found in additional 3 cases. Progressive radiolucent lines in zone 1 and 7 according to Gruen were observed in one case. Postoperative local and general complications were seen as two subfascial hematomas, two single dislocations, two recurrent dislocations of the hip prosthesis, 6 lesions of the sciatic nerve (one persisting), two deep venous thrombosis, two pneumonias, and one lethal pulmonary embolism. A stable proximal fixation was achieved in 242 of 250 cases (96.8%). CONCLUSION: The results of this study using the Vektor-Titan stem in cementless total hip arthroplasty showed that the principle of proximal fixation was optimized. Long term follow-up studies are needed to confirm these good results. PMID- 15792201 TI - Pyomyositis. A literature review. AB - Pyomyositis is a primary bacterial infection of the skeletal muscles. It is known as tropical myositis as most cases have been reported in patients living in tropical areas and, until recently, it was considered unusual in the temperate zone. The condition is usually diagnosed late, and for this reason it is followed by an increased morbidity and sometimes a significant mortality rate. A review of the literature indicates a possibly rising incidence in non-tropical areas. This condition should be considered in patients with muscle pain, fever, leucocytosis, elevated ESR and CRP, in tropical and non-tropical countries, in previously healthy patients as well as in patients suffering other health problems, particularly HIV infection. An MRI investigation should be performed early as it is the imaging technique of choice during the first stage of the disease. A case of a primary iliopsoas pyomyositis diagnosed during the initial stage using MRI is presented as an illustration. PMID- 15792202 TI - Plate fixation of middle-third fractures of the clavicle in the semi-professional athlete. AB - From 1995 to 2003, 39 semi-professional athletes (34 men and 5 women) aged 17-34 years (mountain bike racers, soccer players, swimmers and cyclists) with a displaced fracture of the middle-third of the clavicle were treated operatively using rigid plate fixation within one week after the initial trauma. In 90% of the athletes, radiographic union was achieved after 12 weeks. At 6 weeks post operatively the mean Constant score was 88 points, the mean visual analogue pain score was 3 out of 10 and the average time for return to sports was 45 days. Seventy-five percent of the patients were very satisfied with the end-result and 95% would chose to have the same operation again. In 5% of the cases the end result was unsatisfactory. The most frequent postoperative complication was wound infection (18% of the cases). Nevertheless, this did not affect the outcome. Other postoperative complications in our athletic population included refracture (5%), transient neurological complications (7%) and non-union (5%), which were equally low in the literature. These data suggest that rigid plate fixation of middle-third clavicle fractures gives good results in the semi-professional athlete and may result in early return to sports activity, at the expense of a significant risk for complications, which would not be considered acceptable in patients with lower functional demands. PMID- 15792203 TI - A radiological study of the intraprosthetic movements of the bipolar shoulder replacement in rheumatoid arthritis. AB - A radiological study of the intraprosthetic movements of a bipolar shoulder replacement was undertaken in 25 shoulders in 20 patients with rheumatoid arthritis and rotator cuff arthropathy. No significant difference was found between intraprosthetic movement and shell - glenoid movement. Some shoulders were found to exhibit paradoxical movement at the intraprosthetic interface. Repeating the measurements after an interval of three years in a subgroup of 12 shoulders showed a significant reduction in intraprosthetic movement over this time interval. In conclusion, movement of the bipolar shoulder prosthesis in shoulders affected by rheumatoid arthritis was preserved up to eight years from operation. Intraprosthetic movement was independent from shell - glenoid movement and paradoxical to glenohumeral movement. PMID- 15792204 TI - Anastomosis between the median and ulnar nerve in the forearm. An anatomic study and literature review. AB - Anastomosis between the median and ulnar nerve in the forearm has been shown to be of clinical significance. We aimed to determine the presence of median to ulnar nerve communications in the forearm of the Greek population by anatomical studies. At the same time we defined the types and patterns of the anastomoses found and compared them to those reported in similar studies that were retrieved after a wide review of the literature. One hundred and sixty three forearms from 100 cadavers (53 males, 47 females, 25-91 years old) were carefully dissected to observe median and ulnar nerve communication. The anastomosis was found in 10 cadavers; it was bilateral in 4 and unilateral in 6, on the right side in four and on the left side in two. It occurred in 7 of the 53 male cadavers (14%) and in 3 of the 47 females (6.5%). Overall, the anastomosis was found in 14 of the 163 forearms (8.6%). No case of ulnar to median nerve anastomosis in the forearm was found in anatomical examination. PMID- 15792205 TI - Treatment of distal radial fractures with grafting and K-wiring. AB - We present a series of 10 distal radius fractures treated with either bone substitute or xenograft, and Kapandji's technique (intra-focal threaded Kirschner wires which lever on the graft before penetrating the opposite diaphyseal cortex). The Kirschner wires are buried. Long-term follow-up (one to five years, mean 2.8 years) shows very little secondary displacement (up to 3 mm loss of radial height on the antero-posterior views and none on the sagittal views), excellent tolerability and integration of substitute and good function. The above operative technique has the advantage to allow immediate mobilisation of the wrist. PMID- 15792206 TI - Epidemiology and outcome of complex pelvic injury. AB - Soft tissue injuries associated with pelvic fractures are often responsible for compromised haemodynamics. The objective of this study was to clarify what parameters determine patient outcome. In a cohort study, all patients with a pelvic fracture treated between 1991 and 2001 at a Level I trauma center were analysed for associated intrapelvic injuries, classification, severity of trauma, type of intervention and outcome. Of 552 patients with a pelvic fracture who entered the study, 15.5% presented with associated intrapelvic injuries secondary to the fracture (group I). A subgroup of patients with lacerations of branches of the iliac artery was identified as being at high risk for lethal outcome; they represented 4.3% of all patients with pelvic fracture (group II). The overall mortality reached 4.4%; it increased in group I to 15.5%, and in group II to 33.3%. In the subgroup with pelvic arterial haemorrhage (group II), the severity of injury, the proportion of multiple injured patients, the prevalence of unstable fractures and the incidence of sepsis were significantly increased. The only predictive factor for outcome was the amount of blood transfused, suggesting that fast elimination of the bleeding source decides about patient survival. PMID- 15792207 TI - Comparison of outcomes following uncemented hemiarthroplasty and dynamic hip screw in the treatment of displaced subcapital hip fractures in patients aged greater than 70 years. AB - As health care costs increase, evaluating treatment methods in femoral neck fractures to determine the most effective treatment paradigm will become increasingly important. The current study compared two methods of treatment in similar cohorts of displaced femoral neck fractures. One hundred and twenty two patients were randomly assigned to two groups: In Group A, 62 patients were treated with a hemiarthroplasty. In group B, 60 patients were treated with dynamic screw fixation. Patients were evaluated at a minimum 3 year follow-up. Using the Matta functional hip score, 42% of group A and 70% of group B had good to excellent results. This difference was significant (p = 0.004). A significant agreement between physician assessment using the Matta score, and patient perception of outcome using the SF-36 scale was demonstrated (r = 0.64). No statistical difference between groups for revision surgery existed. Both physician based and patient based outcome scores favour retention and internal fixation of the femoral head in this cohort of patients at a short-term follow up. PMID- 15792208 TI - Wound infection in the management of hip fractures: a comparison between low molecular weight heparin and mechanical prophylaxis. AB - A review of 205 consecutive patients was carried out to determine the association of low molecular weight heparin (LMWH) and the development of wound infection in patients having undergone surgery for a femoral neck fracture: 114 patients treated with LMW Heparin (Group A) and 91 patients with mechanical prophylaxis (Group B). The wounds were assessed using the ASEPSIS Score. Deep vein thrombosis (DVT) and pulmonary embolism (PE) were also noted. Twenty-two patients (19%) in group A developed infection; 9 patients (8%) showed severe infection. Eight patients (8%) in group B developed infection; one patient (1%) showed severe infection. The differences between these two groups regarding infection (p < 0.034) and severity of the infection (p < 0.001) were statistically significant. None of the patients developed PE; however 9 patients were diagnosed with a DVT. Based on these findings, it appears that the use of LMWH for DVT prophylaxis may increase the likelihood of developing a severe wound infection. PMID- 15792209 TI - Component position following total hip arthroplasty through a miniinvasive posterolateral approach. AB - Implant malposition and poor fixation are potential risks of compromising long term results after total hip arthroplasty performed with a minimally invasive technique. Between September 2000 and February 2002, 120 cemented primary total hip arthroplasties were performed at the authors' institutions in patients with primary osteoarthritis of the hip and with BMI lower than 35. In 60 of these cases selected at random, a posterolateral incision no longer than 10 cm was used. The other 60 THA's were performed through a standard posterolateral approach. The inclination and anteversion of the cup and the position of the femoral stem were assessed on radiographs and statistically evaluated. In the miniinvasive group, the average inclination angle of the cup was 42.3 degrees (range: 36 to 52 degrees) and the anteversion angle 13.6 degrees (range: 6 to 29 degrees). The coronal alignment of the femoral component was within 3 degrees of neutral in 54 cases (90.0%). Following conventional implantation in the other group, the average cup inclination angle was 42.4 degrees (range: 35 to 50 degrees) and the anteversion angle 13.6 degrees (range: 8 to 24 degrees. A total of 53 stems (88.3 %) were implanted optimally. Statistical analysis found no significant difference between the two groups regarding components position. These findings suggest that using a smaller posterolateral incision as was done in this study does not introduce a potential risk of compromising long-term results. PMID- 15792210 TI - Assessment of femoral component migration in total hip arthroplasty: digital measurements compared to RSA. AB - The aim of this study was to determine the accuracy of the software system "Dusseldorf Migration Analysis - Femoral Component Analysis" (DMA-FCA) in measuring stem migration in total hip arthroplasty (THA) on digitised anteroposterior radiographs of the pelvis. Bony and implant landmarks on two consecutive radiographs were used for measurements of subsidence and varus-valgus tilt. The accuracy of the method was determined by reference to radiostereometric measurements (RSA). Using specific comparability limits, comparability analysis of radiographs with respect to femoral positioning is possible with DMA. DMA-FCA and RSA measurements were performed after cementless THR in a population of 60 patients aged 38 to 69 years. With a Cronbach's alpha-index of 0.89 and 0.99 for subsidence and 0.90 and 0.98 for classic varus-valgus-tilt, the intraobserver and interobserver reliability for the DMA-FCA-method was calculated as good. Using RSA as reference method, the accuracy of DMA-FCA was calculated to be 2.51 mm for subsidence and 2.49 degrees for varus-valgus-tilt (95% confidence interval). Without comparison to RSA, DMA measured 1.94 mm for subsidence and 2.35 degrees for varus-valgus-tilt. Based on a comparison with RSA, our results show lower accuracy for DMA-FCA than for EBRA-FCA, but DMA-FCA is easier to use in everyday clinical practice. It is hoped that the use of digital measuring methods such as DMA will become standard for long-term observation and will be integrated into clinical routine in the context of quality assurance of THR. PMID- 15792211 TI - Arthrodesis of the first metatarsophalangeal joint using convex and concave drills. A report on 50 cases. AB - The authors report their results after arthrodesis of the first metatarsophalangeal joint (MP) performed in 50 cases including 18 rheumatoid feet, 18 major hallux valgus deformities and 14 recurrent deformities. Preparation of the bone ends for arthrodesis was achieved using convex and concave drills and fixation was done with an axial screw. The patients were evaluated after an average of 12 months, using Kitaoka's score. Eighty per cent were very satisfied, 16% were satisfied and 4% were dissatisfied. Ninety six percent reported no pain over the MP joint of the hallux, and none complained of metatarsalgia. Return to activities was normal in 80% of cases. Radiographs showed non-union in six cases; they did not show interphalangeal osteoarthritis in any instance. PMID- 15792212 TI - Instrumented in situ posterolateral fusion for low-grade lytic spondylolisthesis in adults. AB - Lytic olisthesis is an accepted cause of low back pain and may also be associated with leg pain. When conservative treatment fails, operative management with fusion and if necessary decompression is considered. Most reports regarding surgical management in the literature relate to children or adolescents. This paper reviews 75 adult cases treated by in situ pedicular fixation and posterolateral fusion with simultaneous decompression in 55 cases. The mean follow-up was 72 months. Ninety-six percent of the cases had a satisfactory clinical outcome, and this correlates closely with a solid fusion. This approach therefore appears to be safe and successful in the treatment of symptomatic low grade lytic spondylolisthesis in adults. PMID- 15792213 TI - The removal of percutaneous Kirschner wires used in the stabilisation of fractures in children. AB - Kirschner wires are commonly used to stabilise fractures in children. There is a wide range of practice across the United Kingdom with regard to their removal, with some units routinely readmitting children for removal of wires under a general anaesthetic and others leaving the wires long and extracting them in the outpatient clinic. In a prospective study performed at The Barts and Royal London National Health Service Trust between November 1997 and December 1999, the acceptability and problems associated with the latter approach were assessed. A total of 203 percutaneous Kirschner wires were used to stabilise 119 fractures following closed reduction in 113 children. The majority of the fractures were in the upper limb and the children ranged in age from five months to fifteen years and five months. The Kirschner wires were subsequently removed in clinic without anaesthetic. Details of wire placement and complications associated with wire use and their removal were recorded. The most common complication seen was overgranulation at the entry point of the wire (5.4%). In two cases, wires had migrated below the skin surface, requiring local anaesthetic infiltration to enable retrieval. Using a visual analogue pain score, the mean pain score immediately after wire removal was 2.8 out of a maximum of 10, reducing to 0.9 after one minute. It is concluded that K-wires can be removed safely without anaesthetic in the outpatient clinic and that this is tolerated well by children. PMID- 15792214 TI - Cyclic administration of pamidronate to treat osteoporosis in children with cerebral palsy or a neuromuscular disorder: a clinical study. AB - Eighteen osteoporotic (Z-score at or below 2.5) nonambulatory children with cerebral palsy or a neuromuscular disorder received cyclic intravenous administration of pamidronate. One year after treatment, bone densitometry showed an improvement in all patients: a mean increase of 13% +/- 15% standard deviation (SD) (p < 10(-5)) on the global data, and a mean increase of 27% +/- 15% (p < 10( 5)) on the most significant area for each individual patient. Clinical improvement was found in all patients, with a decrease in pain on manipulation, and no new fractures. No major adverse effects were reported. Cyclic intravenous administration of pamidronate is a useful tool in the treatment of osteoporosis in these children. PMID- 15792215 TI - Treatment of recurrent dislocation associated with impingement after revision total hip arthroplasty. AB - The authors report a case of total hip replacement in which, following several revision operations ending up in a Girdlestone arthroplasty, a patient presented recurrent episodes of posterior dislocation of his total hip prosthesis after revision using an antiprotrusio cage. The main causes for dislocation were anterior impingement of the neck against the anterior wall of the antiprotrusio cage and slackness of the periarticular musculature, due to shortening and reduced femoral offset. In order to avoid further major reconstruction and minimise the risk of infection recurrence, a limited revision operation was performed to address the factors responsible for the instability: the anterior wall of the antiprotrusio cage was removed and an extension module was added to the femoral component. Good stability of the arthroplasty was achieved with this limited procedure. PMID- 15792216 TI - Recurrent torticollis secondary to Langerhans cell histiocytosis: a case report. AB - Torticollis is a common clinical sign encountered by pediatricians and orthopaedic surgeons in a wide spectrum of childhood conditions ranging from benign to life-threatening. We report the case of a child with recurrent torticollis caused by Langerhans Cell Histiocytosis (LCH). The patient was a 1 year-old boy with recurrent torticollis, followed by a painless swelling over the right temporal bone. The diagnosis was confirmed by an open biopsy of the calvarial lesion. As LCH is a very rare cause of torticollis it was not considered in the initial differential by the primary care physicians and the diagnosis was delayed about 4 months. The patient received chemotherapy with steroids and etoposide for 52 weeks. He showed complete regression of the sign and imaging tests at the end of treatment were normal. No relapse of symptoms occurred during a follow-up period of 2 years. The rarity of this disease as well as the site and form of presentation are emphasised to alert physicians for an early diagnostic evaluation, which is important to prevent neurological lesions and other late complications. PMID- 15792217 TI - Chronic rupture of the pectoralis major muscle: report of two cases. AB - Rupture of the pectoralis major muscle is relatively rare. The authors present two cases of chronic rupture in which direct suturing was not possible; reconstruction was possible using a bone-patellar tendon autograft. They conclude that this surgical technique may be useful in some cases of chronic ruptures of the pectoralis major muscle in which direct tendon suturing is impossible. PMID- 15792218 TI - Traumatic anterior dislocation of the hip joint with posterior acetabular wall fracture. AB - The incidence of traumatic hip dislocation has increased in recent years as a result of high-energy trauma. Anterior hip dislocation forms less than 10-15% of all traumatic hip dislocations. Only a few case reports describe anterior dislocation along with acetabular fractures. The acetabular fracture involved the anterior wall or column in all such cases. We describe a rare case in which anterior superior dislocation of the hip was associated with a large fracture fragment of theposterior acetabular rim and adjacent wall. PMID- 15792219 TI - Extraskeletal osteochondroma of the thigh: a case report. AB - A case of histopathologically proven extraskeletal osteochondroma of the thigh is presented along with its radiographic, CT and MRI findings. This is the first such case reported, to the best of our knowledge. The diagnosis of extraskeletal osteochondroma should be considered when a discrete ossified mass is localised in the soft tissue. PMID- 15792220 TI - Infection of a total knee prosthesis by Candida glabrata: a case report. PMID- 15792221 TI - Primary haemangiopericytoma of the tibia. Treatment by en bloc resection and bone transportation. A case report. PMID- 15792222 TI - Statistical analysis of diversification with species traits. AB - Testing whether some species traits have a significant effect on diversification rates is central in the assessment of macroevolutionary theories. However, we still lack a powerful method to tackle this objective. I present a new method for the statistical analysis of diversification with species traits. The required data are observations of the traits on recent species, the phylogenetic tree of these species, and reconstructions of ancestral values of the traits. Several traits, either continuous or discrete, and in some cases their interactions, can be analyzed simultaneously. The parameters are estimated by the method of maximum likelihood. The statistical significance of the effects in a model can be tested with likelihood ratio tests. A simulation study showed that past random extinction events do not affect the Type I error rate of the tests, whereas statistical power is decreased, though some power is still kept if the effect of the simulated trait on speciation is strong. The use of the method is illustrated by the analysis of published data on primates. The analysis of these data showed that the apparent overall positive relationship between body mass and species diversity is actually an artifact due to a clade-specific effect. Within each clade the effect of body mass on speciation rate was in fact negative. The present method allows to take both effects (clade and body mass) into account simultaneously. PMID- 15792223 TI - Evolution of migration under kin selection and local adaptation. AB - We present here a stochastic two-locus, two-habitat model for the evolution of migration with local adaptation and kin selection. One locus determines the migration rate while the other causes local adaptation. We show that the opposing forces of kin competition and local adaptation can lead to the existence of one or two convergence stable migration rates, notably depending on the recombination rate between the two loci. We show that linkage between migration and local adaptation loci has two antagonist effects: when linkage is tight, cost of local adaptation increases, leading to smaller equilibrium migration rates. However, when linkage is tighter, the population structure at the migration locus tends to be very high because of the indirect selection, and thus equilibrium migration rates increases. This result, qualitatively different from results obtained with other models of migration evolution, indicates that ignoring drift or the detail of the genetic architecture may lead to incorrect conclusions. PMID- 15792224 TI - Gene tree distributions under the coalescent process. AB - Under the coalescent model for population divergence, lineage sorting can cause considerable variability in gene trees generated from any given species tree. In this paper, we derive a method for computing the distribution of gene tree topologies given a bifurcating species tree for trees with an arbitrary number of taxa in the case that there is one gene sampled per species. Applications for gene tree distributions include determining exact probabilities of topological equivalence between gene trees and species trees and inferring species trees from multiple datasets. In addition, we examine the shapes of gene tree distributions and their sensitivity to changes in branch lengths, species tree shape, and tree size. The method for computing gene tree distributions is implemented in the computer program COAL. PMID- 15792225 TI - Constrained camouflage facilitates the evolution of conspicuous warning coloration. AB - The initial evolution of aposematic and mimetic antipredator signals is thought to be paradoxical because such coloration is expected to increase the risk of predation before reaching a stage when predators associate it effectively with a defense. We propose, however, that constraints associated with the alternative strategy, cryptic coloration, may facilitate the evolution of antipredator signals and thus provide a solution for the apparent paradox. We tested this hypothesis first using an evolutionary simulation to study the effect of a constraint due to habitat heterogeneity, and second using a phylogenetic comparison of the Lepidoptera to investigate the effect of a constraint due to prey motility. In the evolutionary simulation, antipredator warning coloration had an increased probability to invade the prey population when the evolution of camouflage was constrained by visual difference between microhabitats. The comparative study was done between day-active lepidopteran taxa, in which camouflage is constrained by motility, and night-active taxa, which rest during the day and are thus able to rely on camouflage. We compared each of seven phylogenetically independent day-active groups with a closely related nocturnal group and found that antipredator signals have evolved at least once in all the diurnal groups but in none of their nocturnal matches. Both studies lend support to our idea that constraints on crypsis may favor the evolution of antipredator warning signals. PMID- 15792226 TI - Loss of gametophytic self-incompatibility with evolution of inbreeding depression. AB - Gametophytic self-incompatibility (SI) in plants is a widespread mechanism preventing self-fertilization and the ensuing inbreeding depression, but it often evolves to self-compatibility. We analyze genetic mechanisms for the breakdown of gametophytic SI, incorporating a dynamic model for the evolution of inbreeding depression allowing for partial purging of nearly recessive lethal mutations by selfing, and accounting for pollen limitation and sheltered load linked to the S locus. We consider two mechanisms for the breakdown of gametophytic SI: a nonfunctional S-allele and an unlinked modifier locus that inactivates the S locus. We show that, under a wide range of conditions, self-compatible alleles can invade a self-incompatible population. Conditions for invasion are always less stringent for a nonfunctional S-allele than for a modifier locus. The spread of self-compatible genotypes is favored by extremely high or low selfing rates, a small number of S-alleles, and pollen limitation. Observed parameter values suggest that the maintenance of gametophytic SI is caused by a combination of high inbreeding depression in self-incompatible populations coupled with intermediate selfing rates of the self-compatible genotypes and sheltered load linked to the S-locus. PMID- 15792227 TI - Plant genetic determinants of arthropod community structure and diversity. AB - To test the hypothesis that genes have extended phenotypes on the community, we quantified how genetic differences among cottonwoods affect the diversity, abundance, and composition of the dependent arthropod community. Over two years, five major patterns were observed in both field and common-garden studies that focused on two species of cottonwoods and their naturally occurring F1 and backcross hybrids (collectively referred to as four different cross types). We did not find overall significant differences in arthropod species richness or abundance among cottonwood cross types. We found significant differences in arthropod community composition among all cross types except backcross and narrowleaf cottonwoods. Thus, even though we found similar richness among cross types, the species that composed the community were significantly different. Using vector analysis, we found that the shift in arthropod community composition was correlated with percent Fremont alleles in the host plant, which suggests that the arthropod community responds to the underlying genetic differences among trees. We found 13 arthropod species representing different trophic levels that were significant indicators of the four different cross types. Even though arthropod communities changed in species composition from one year to the next, the overall patterns of community differences remained remarkably stable, suggesting that the genetic differences among cross types exert a strong organizing influence on the arthropod community. Together, these results support the extended phenotype concept. Few studies have observationally and experimentally shown that entire arthropod communities can be structured by genetic differences in their host plants. These findings contribute to the developing field of community genetics and suggest a strategy for conserving arthropod diversity by promoting genetic diversity in their host plants. PMID- 15792228 TI - Host-parasite and genotype-by-environment interactions: temperature modifies potential for selection by a sterilizing pathogen. AB - Parasite-mediated selection is potentially of great importance in modulating genetic diversity. Genetic variation for resistance, the fuel for natural selection, appears to be common in host-parasite interactions, but responses to selection are rarely observed. In the present study, we tested whether environmental variation could mediate infection and determine evolutionary outcomes. Temperature was shown to dramatically alter the potential for parasite mediated selection in two independent laboratory infection experiments at four temperatures. The bacterial parasite, Pasteuria ramosa, was extremely virulent at 20 degrees C and 25 degrees C, sterilizing its host, Daphnia magna, so that females often never produced a single brood. However, at 10 degrees C and 15 degrees C, the host-parasite interaction was much more benign, as nearly all females produced broods before becoming sterile. This association between virulence and temperature alone could stabilize coexistence and lead to the maintenance of diversity, because it would weaken parasite-mediated selection during parts of the season. Additionally, highly significant genotype-by environment interactions were found, with changes in clone rank order for infection rates at different temperatures. Our results clearly show that the outcome of parasite-mediated selection in this system is strongly context dependent. PMID- 15792229 TI - Quantitative trait loci affecting delta13C and response to differential water availibility in Arabidopsis thaliana. AB - Phenotypic plasticity is an important response mechanism of plants to environmental heterogeneity. Here, we explored the genetic basis of plastic responses of Arabidopsis thaliana to water deficit by experimentally mapping quantitative trait loci (QTL) in two recombinant inbred populations (Cvi x Ler and Ler x Col). We detected genetic variation and significant genotype-by environment interactions for many traits related to water use. We also mapped 26 QTL, including six for carbon isotope composition (delta13C). Negative genetic correlations between fruit length and fruit production as well as between flowering time and branch production were corroborated by QTL colocalization, suggesting these correlations are due to pleiotropy or physical linkage. Water limited plants were more apically dominant with greater root:shoot ratios and higher delta13C (higher water-use efficiency) when compared to well-watered plants. Many of the QTL effects for these traits interacted significantly with the irrigation treatment, suggesting that the observed phenotypic plasticity is genetically based. We specifically searched for epistatic (QTL-QTL) interactions using a two-dimensional genome scan, which allowed us to detect epistasis regardless of additive genetic effects. We found several significant QTL-QTL interactions including three that exhibited environmental dependence. These results provide preliminary evidence for proposed genetic mechanisms underlying phenotypic plasticity. PMID- 15792230 TI - Conspecific sperm precedence in two species of tropical sea urchins. AB - Conspecific sperm precedence occurs when females are exposed to sperm from males of multiple species, but preferentially use sperm of a conspecific. Conspecific sperm precedence and its mechanisms have been documented widely in terrestrial species, in which complex female behaviors or reproductive tract morphologies can allow many opportunities for female choice and sperm competition, however, the opportunity for conspecific sperm precedence in free spawning marine invertebrates has been largely ignored. Two sea urchin species, Echinometra oblonga and E. sp. C, have high levels of interspecific fertilization in no choice lab crosses, but no natural hybrids have been found. We performed competitive fertilization assays to test for conspecific sperm precedence and found that eggs of both species showed a marked preference for conspecific sperm when fertilized with heterospecific sperm mixtures. Strong rejection of heterospecific sperm would not have been predicted from no-choice assays and helps explain the lack of natural hybrids. We also found significant variation in hybridization success among crosses. Conspecific sperm precedence in free spawning invertebrates shows that the simple surfaces of eggs and sperm provide ample opportunity for egg choice and sperm competition even in the absence of intricate behavior or complex reproductive morphologies. PMID- 15792231 TI - Male-by-female interactions influence fertilization success and mediate the benefits of polyandry in the sea urchin Heliocidaris erythrogramma. AB - Numerous studies have reported that females benefit from mating with multiple males (polyandry) by minimizing the probability of fertilization by genetically incompatible sperm. Few, however, have directly attributed variation in female reproductive success to the fertilizing capacity of sperm. In this study we report on two experiments that investigated the benefits of polyandry and the interacting effects of males and females at fertilization in the free-spawning Australian sea urchin Heliocidaris erythrogramma. In the first experiment we used a paired (split clutch) experimental design and compared fertilization rates within female egg clutches under polyandry (eggs exposed to the sperm from two males simultaneously) and monandry (eggs from the same female exposed to sperm from each of the same two males separately). Our analysis revealed a significant fertilization benefit of polyandry and strong interacting effects of males and females at fertilization. Further analysis of these data strongly suggested that the higher rates of fertilization in the polyandry treatment were due to an overrepresentation of fertilizations due to the most compatible male. To further explore the interacting effects of males and females at fertilization we performed a second factorial experiment in which four males were crossed with two females (in all eight combinations). In addition to confirming that fertilization success is influenced by male x female interactions, this latter experiment revealed that both sexes contributed significant variance to the observed patterns of fertilization. Taken together, these findings highlight the importance of male x female interactions at fertilization and suggest that polyandry will enable females to reduce the cost of fertilization by incompatible gametes. PMID- 15792232 TI - Fine scale endemism on coral reefs: archipelagic differentiation in turbinid gastropods. AB - The perceived wide geographic range of organisms in the sea, facilitated by ready dispersal of waterborne dispersal stages, is a challenge for hypotheses of marine speciation but a boon to efforts of marine conservation. Wide species ranges are especially striking in the reef-rich Indo-west Pacific, the largest and most diverse marine biogeographic region, extending across half the planet. The insular marine biota of the tropical Pacific is characterized by wide-ranging species and provides the most striking examples of long distance dispersal, with endemism largely confined to the most remote island groups. Here we show that the gastropod Astralium "rhodostomum" has developed endemic clades on almost every Pacific archipelago sampled, a pattern unprecedented in marine biogeography, and reminiscent of the terrestrial biota of oceanic islands. Mitochondrial DNA sequences indicate that this species-complex is comprised of at least 30 geographically isolated clades, separated by as little as 180 km. Evidence suggests that such fine scale endemism and high diversity is not exceptional, but likely characterizes a substantial fraction of the reef biota. These results imply that (1) marine speciation can regularly occur over much finer spatial scales than generally accepted, (2) the diversity of coral reefs is even higher than suggested by morphology-based estimates, and (3) conservation efforts need to focus at the archipelagic level in the sea as on land. PMID- 15792233 TI - Rates of divergence in gene expression profiles of primates, mice, and flies: stabilizing selection and variability among functional categories. AB - The extent to which natural selection shapes phenotypic variation has long been a matter of debate among those studying organic evolution. We studied the patterns of gene expression polymorphism and divergence in several datasets that ranged from comparisons between two very closely related laboratory strains of mice to comparisons across a considerably longer time scale, such as between humans and chimpanzees, two species of mice, and two species of Drosophila. The results were analyzed and interpreted in view of neutral models of phenotypic evolution. Our analyses used a number of metrics to show that most mRNA levels are evolutionary stable, changing little across the range of taxonomic distances compared. This implies that, overall, widespread stabilizing selection on transcription levels has prevented greater evolutionary changes in mRNA levels. Nevertheless, the range of rates of divergence is large with highly significant differences in the rate and patterns of transcription divergence across functional classes defined on the basis of the gene ontology annotation (primates and mice datasets) or on the basis of the pattern of sex-biased gene expression (Drosophila). Moreover, rates of divergence of sex-biased genes in the contrast between Drosophila species show a distinct pattern from that observed in the contrast between populations of D. melanogaster. Hence, we discuss the time scale of the changes observed and its consequences for the relationship between variation in gene expression within and between species. Finally, we argue that differences in mRNA levels of the magnitudes observed herein could be explained by a remarkably small number of generations of directional selection. PMID- 15792234 TI - Sexual selection, genetic architecture, and the condition dependence of body shape in the sexually dimorphic fly Prochyliza xanthostoma (Piophilidae). AB - The hypothesis that sexual selection drives the evolution of condition dependence is not firmly supported by empirical evidence, and the process remains poorly understood. First, even though sexual competition typically involves multiple traits, studies usually compare a single sexual trait with a single "control" trait, ignoring variation among sexual traits and raising the possibility of sampling bias. Second, few studies have addressed the genetic basis of condition dependence. Third, even though condition dependence is thought to result from a form of sex-specific epistasis, the evolution of condition dependence has never been considered in relation to intralocus sexual conflict. We argue that condition dependence may weaken intersexual genetic correlations and facilitate the evolution of sexual dimorphism. To address these questions, we manipulated an environmental factor affecting condition (larval diet) and examined its effects on four sexual and four nonsexual traits in Prochyliza xanthostoma adults. As predicted by theory, the strength of condition dependence increased with degree of exaggeration among male traits. Body shape was more condition dependent in males than in females and, perhaps as a result, genetic and environmental effects on body shape were congruent in males, but not in females. However, of the four male sexual traits, only head length was significantly larger in high-condition males after controlling for body size. Strong condition dependence was associated with reduced intersexual genetic correlation. However, homologous male and female traits exhibited correlated responses to condition, suggesting an intersexual genetic correlation for condition dependence itself. Our findings support the role of sexual selection in the evolution of condition dependence, but reveal considerable variation in condition dependence among sexual traits. It is not clear whether the evolution of condition dependence has mitigated or exacerbated intralocus sexual conflict in this species. PMID- 15792235 TI - Polyandry promotes enhanced offspring survival in decorated crickets. AB - Although female multiple mating is ubiquitous in insects, its adaptive significance remains poorly understood. Benefits to multiple mating can accrue via direct material benefits, indirect genetic benefits, or both. We investigated the effects of multiple mating in the decorated cricket, Gryllodes sigillatus, by simultaneously varying the number of times that females mated and the number of different males with which they mated, measuring aspects of female fecundity and elements of offspring performance and viability. Multiple matings resulted in enhanced female fitness relative to single matings when females mated with different partners, but not when females mated repeatedly with the same male. Specifically, polyandrous females produced significantly more offspring surviving to reproductive maturity than did monogamous females mating once or mating repeatedly with the same male. These results suggest that the benefit females gain from multiple mating is influenced primarily by genetic and not material benefits. PMID- 15792236 TI - The transition to social inbred mating systems in spiders: role of inbreeding tolerance in a subsocial predecessor. AB - The social spiders are unusual among cooperatively breeding animals in being highly inbred. In contrast, most other social organisms are outbred owing to inbreeding avoidance mechanisms. The social spiders appear to originate from solitary subsocial ancestors, implying a transition from outbreeding to inbreeding mating systems. Such a transition may be constrained by inbreeding avoidance tactics or fitness loss due to inbreeding depression. We examined whether the mating system of a subsocial spider, in a genus with three social congeners, is likely to facilitate or hinder the transition to inbreeding social systems. Populations of subsocial Stegodyphus lineatus are substructured and spiders occur in patches, which may consist of kin groups. We investigated whether male mating dispersal prevents matings within kin groups in natural populations. Approximately half of the marked males that were recovered made short moves (< 5m) and mated within their natal patch. This potential for inbreeding was counterbalanced by a relatively high proportion of immigrant males. In mating experiments, we tested whether inbreeding actually results in lower offspring fitness. Two levels of inbreeding were tested: full sibling versus non-sib matings and matings of individuals within and between naturally occurring patches of spiders. Neither full siblings nor patch mates were discriminated against as mates. Sibling matings had no effect on direct fitness traits such as fecundity, hatching success, time to hatching and survival of the offspring, but negatively affected offspring growth rates and adult body size of both males and females. Neither direct nor indirect fitness measures differed significantly between within patch and between-patch pairs. We tested the relatedness between patch mates and nonpatch mates using DNA fingerprinting (TE AFLP). Kinship explained 30% of the genetic variation among patches, confirming that patches are often composed of kin. Overall, we found limited male dispersal, lack of kin discrimination, and tolerance to low levels of inbreeding. These results suggest a history of inbreeding which may reduce the frequency of deleterious recessive alleles in the population and promote the evolution of inbreeding tolerance. It is likely that the lack of inbreeding avoidance in subsocial predecessors has facilitated the transition to regular inbreeding social systems. PMID- 15792237 TI - Countergradient variation in the sexual coloration of guppies (Poecilia reticulata): drosopterin synthesis balances carotenoid availability. AB - Trinidad guppies (Poecilia reticulata) are distributed along an environmental gradient in carotenoid availability that limits the carotenoid content of the orange spots of males. The amount of synthetic red pteridines (drosopterins) in the orange spots covaries with the carotenoid content, such that the ratio of the two types of pigments is roughly conserved across streams. Carotenoids and drosopterins have different spectral properties and thus the ratio of the two types of pigments affects the shape of the orange spot reflectance spectrum. Geographic conservation of the carotenoid:drosopterin ratio suggests that males may be under selection to maintain a particular hue. We tested this hypothesis by comparing the pigmentation and coloration of guppies from six streams in the field to that of second-generation descendants of the same populations raised on three dietary carotenoid levels in the laboratory. The results show clearly that the geographic variation in drosopterin production is largely genetic and that the hue of the orange spots is conserved among populations in the field, relative to the laboratory diet groups. This is a countergradient pattern because genetic differences between populations in drosopterin production mask the effect of carotenoid availability on the hue of the orange spots. The potential for countergradient sexual selection to contribute to reproductive isolation between populations is discussed. PMID- 15792238 TI - Growth rate correlates to individual heterozygosity in the European eel, Anguilla anguilla L. AB - Heterozygosity-fitness correlations (HFCs) have been reported in populations of many species. We provide evidence for a positive correlation between genetic variability and growth rate at 12 allozyme loci in a catadromous marine fish species, the European eel (Anguilla anguilla L.). More heterozygous individuals show a significantly higher length and weight increase and an above average condition index in comparison with more homozygous individuals. To a lesser extent, six microsatellite loci show a similar pattern, with positive but not significant correlations between heterozygosity and growth rate. The HFCs observed could be explained by an effect of either direct allozyme over-dominance or associative overdominance. Selection affecting some of the allozyme loci would explain the greater strength of the HFCs found at allozymes in comparison with microsatellites and the lack of correlation between MLH at allozymes and MLH at microsatellites. Associative overdominance (where allozyme loci are merely acting as neutral markers of closely linked fitness loci) might provide an explanation for the HFCs if we consider that allozyme loci have a higher chance than microsatellites to be in linkage disequilibrium with fitness loci. PMID- 15792239 TI - Ecological adaptation and species recognition drives vocal evolution in neotropical suboscine birds. AB - Given that evolutionary divergence in mating signals leads to reproductive isolation in numerous animal taxa, understanding what drives signal divergence is fundamental to our understanding of speciation. Mating signals are thought to diverge via several processes, including (1) as a by-product of morphological adaptation, (2) through direct adaptation to the signaling environment, or (3) to facilitate species recognition. According to the first two hypotheses, birdsongs diversify in different foraging niches and habitats as a product of selection for optimal morphology and efficient sound transmission, respectively. According to the third hypothesis, they diversify as a result of selection against maladaptive hybridization. In this study I test all three hypotheses by examining the influence of morphology, acoustic environment, and the presence of closely related congeners on song structure in 163 species of antbird (Thamnophilidae). Unlike oscine passerines, these Neotropical suboscines make ideal subjects because they develop their songs without learning. In other words, patterns of vocal divergence are not complicated by cultural evolution. In support of the morphological adaptation hypothesis, body mass correlates with the acoustic frequency of songs, and bill size with temporal patterning. These relationships were robust, even when controlling for phylogenetic inertia using independent contrasts, suggesting that there has been correlated evolution between morphological and acoustic traits. The results also support the acoustic adaptation hypothesis: birds which habitually sing in the understory and canopy produce higher-pitched songs than those that sing in the midstory, suggesting that song structure is related to the sound transmission properties of different habitat strata. Finally, the songs of sympatric pairs of closely related species are more divergent than those of allopatric pairs, as predicted by the species recognition hypothesis. To my knowledge, these data provide the first direct evidence that species recognition and ecological adaptation operate in tandem, and that the interplay between these factors drives the evolution of mating signals in suboscine birds. PMID- 15792240 TI - The probability of parallel evolution. AB - How often will natural selection drive parallel evolution at the DNA sequence level? More precisely, what is the probability that selection will cause two populations that live in identical environments to substitute the same beneficial mutation? Here I show that, under fairly general conditions, the answer is simple: if a wild-type sequence can mutate to n different beneficial mutations, replicate populations will on average fix the same mutation with probability P = 2/(n + 1). This probability, which is derived using extreme value theory, is independent of most biological details, including the length of the gene in question and the precise distribution of fitness effects among alleles. I conclude that the probability of parallel evolution under natural selection is nearly twice as large as that under neutrality. PMID- 15792241 TI - The role of Haldane's rule in sex allocation. AB - Sex allocation theory predicts that parents should bias their reproductive investments toward the offspring sex generating the greatest fitness return. When females are the heterogametic sex (e.g., ZW in butterflies, some lizards, and birds), production of daughters is associated with an increased risk of offspring inviability due to the expression of paternal, detrimental recessives on the Z chromosome. Thus, daughters should primarily be produced when mating with partners of high genetic quality. When female sand lizards (Lacerta agilis) mate with genetically superior males, exhibiting high MHC Class I polymorphism, offspring sex ratios are biased towards daughters, possibly due to recruitment of more Z-carrying oocytes when females have assessed the genetic quality of their partners. If our study has general applicability across taxa, it predicts taxon specific sex allocation effects depending on which sex is the heterogametic one. PMID- 15792242 TI - Rapid and repeated origin of insular gigantism and dwarfism in Australian tiger snakes. AB - It is a well-known phenomenon that islands can support populations of gigantic or dwarf forms of mainland conspecifics, but the variety of explanatory hypotheses for this phenomenon have been difficult to disentangle. The highly venomous Australian tiger snakes (genus Notechis) represent a well-known and extreme example of insular body size variation. They are of special interest because there are multiple populations of dwarfs and giants and the age of the islands and thus the age of the tiger snake populations are known from detailed sea level studies. Most are 5000-7000 years old and all are less than 10,000 years old. Here we discriminate between two competing hypotheses with a molecular phylogeography dataset comprising approximately 4800 bp of mtDNA and demonstrate that populations of island dwarfs and giants have evolved five times independently. In each case the closest relatives of the giant or dwarf populations are mainland tiger snakes, and in four of the five cases, the closest relatives are also the most geographically proximate mainland tiger snakes. Moreover, these body size shifts have evolved extremely rapidly and this is reflected in the genetic divergence between island body size variants and mainland snakes. Within south eastern Australia, where populations of island giants, populations of island dwarfs, and mainland tiger snakes all occur, the maximum genetic divergence is only 0.38%. Dwarf tiger snakes are restricted to prey items that are much smaller than the prey items of mainland tiger snakes and giant tiger snakes are restricted to seasonally available prey items that are up three times larger than the prey items of mainland tiger snakes. We support the hypotheses that these body size shifts are due to strong selection imposed by the size of available prey items, rather than shared evolutionary history, and our results are consistent with the notion that adaptive plasticity also has played an important role in body size shifts. We suggest that plasticity displayed early on in the occupation of these new islands provided the flexibility necessary as the island's available prey items became more depauperate, but once the size range of available prey items was reduced, strong natural selection followed by genetic assimilation worked to optimize snake body size. The rate of body size divergence in haldanes is similar for dwarfs (h(g) = 0.0010) and giants (h(g) = 0.0020-0.0025) and is in line with other studies of rapid evolution. Our data provide strong evidence for rapid and repeated morphological divergence in the wild due to similar selective pressures acting in different directions. PMID- 15792243 TI - Geographical variation in the rate of evolution: effect of available energy or fluctuating environment? AB - In a recent paper, Wright et al. (2003) argue for the hypothesis that greater biologically available energy elevates the rate of molecular evolution. However, their results are also consistent with alternative hypotheses that invoke either environmentally driven variation in effective population sizes, or natural selection, or both. The available energy gradient cited by Wright et al. is linearly correlated with temperature fluctuations, and the observed rate heterogeneity could be a consequence of this environmental variability. The distribution of phylogenetic branch lengths alone is insufficient to distinguish between the hypotheses, and complementary approaches are suggested. PMID- 15792244 TI - Increased rates of molecular evolution in an equatorial plant clade: an effect of environment or phylogenetic nonindependence? AB - A recent study of environmental effects on rates of molecular evolution in the plant subgenus Mearnsia shows that species occurring in more equatorial latitudes have higher rates of substitution in rDNA sequences as compared to their more southerly congeners (Wright et al. 2003). However, we believe that the statistical approach employed by Wright et al. (2003) insufficiently accounts for the phylogenetic nonindependence of the species examined, given that all six equatorial species of Mearnsia form a clade. To distinguish between the effect of latitude and that of phylogenetic nonindependence, we have employed a variety of comparative approaches that use independent contrasts to test for an effect of environment across this entire subgenus. We find very little evidence for an effect of latitude on rate of molecular evolution using these approaches and believe that the shared evolutionary history of the clade is a plausible explanation of the apparent rate difference between equatorial and subequatorial Mearnsia species. PMID- 15792245 TI - Influence of the pore structure on the properties of silica based reversed phase packings for LC. AB - This paper describes the preparation and investigation of new, highly loaded, monomeric, silica based, reversed phase C18 and C30 packings. The influence of pore structure and endcapping on the properties of C18 and C30 packings is described. Using hydrothermal procedures, silicas with predictable pore size (9.3 25.5 nm) and surface area have been prepared. Silylation with long chain silanes substantially alters the pore structure of the silica: pore size and pore volume decrease. A new parameter, the volumetric surface coverage [mm3 x m(-2)] has been introduced. This parameter--calculated from on-column measured porosity data- indicates the pore volume portion occupied by the hydrocarbon chains. Endcapping does not significantly change the pore structure of the bonded phases. The reduced retentions (reduced with respect to unit area: [k/m2])--a good measure for comparing the retention behaviour of packings with different surface areas- are similar for most of the phases, demonstrating good accessibility of the pores for the solutes. Slightly lower retentions were found on the endcapped than on the non-endcapped phases for probes with dense pi-electron system (e.g. polyaromatic hydrocarbons) demonstrating the contribution of silanophilic interactions to the retention. The phases had been successfully used for various demanding separations, e.g. for the separation of flavonoids, carotenoids, resveratrol, and tocopherol isomers, fullerenes, and anions. PMID- 15792246 TI - Direct determination of five fluoroquinolones in chicken whole blood and in veterinary drugs by HPLC. AB - A direct, accurate, and sensitive chromatographic analytical method for the quantitative determination of five fluoroquinolones (enoxacin, ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin) in chicken whole blood is proposed in the present study. For quantitative determination lamotrigine was used as internal standard at a concentration of 20 ng/microL. The developed method was successfully applied to the determination of enrofloxacin, as the main component of commercially available veterinary drugs. Fluoroquinolone antibiotics were separated on an Inertsil (250 x 4 mm) C8, 5 microm, analytical column, at ambient temperature. The mobile phase consisted of a mixture of citric acid (0.4 mol L( 1))-CH3OH-CH3CN (87:9:4% v/v) leading to retention times less than 14 min, at a flow rate 1.4 mL min(-1). UV detection at 275 nm provided limits of detection of 2 ng/mL per 20 microL injected volume for enoxacin, norfloxacin, and ciprofloxacin, 0.4 ng/mL for ofloxacin, and 4 ng/mL for enrofloxacin. Preparation of chicken blood samples is based on the deproteinization with acetonitrile while the pharmaceutical drug was simply diluted with water. Peaks of examined analytes in real samples were identified by means of a photodiode array detector. The method was validated in terms of within-day (n=6) precision and accuracy after chicken whole blood sample deproteinization by CH3CN. Using 50 microL of chicken blood sample, recovery rates at fortification levels of 40, 60, and 80 ng ranged from 86.7% to 103.7%. The applicability of the method was evaluated using real samples from chicken under fluoroquinolone treatment. PMID- 15792247 TI - Quantitative determination of antalarmin, a novel corticotropin-releasing hormone receptor-1 antagonist, in canine plasma by HPLC-MS. AB - A simple and rapid method was developed for the quantitation of antalarmin from plasma using high performance liquid chromatography coupled with electrospray ionization mass spectrometry (ESI/MS). Separation of antalarmin from interfering compounds was achieved using reversed phase chromatography on a C-8 micro-column with an isocratic mobile phase comprised of 80% acetonitrile, 20% water, and 5 mM triethylamine. Detection by ESI/MS was accomplished in positive ion mode using single ion monitoring of the protonated molecular ions of antalarmin and its 13C2 isotopimer. The area ratio of the integrated peaks of interest in the extracted ion chromatogram was used for quantitation. The lower limit of detection was 1 picogram (pg) and the quantitation showed a linear response up to 4 nanograms loaded on column. To achieve acceptable accuracy at or around the limit of quantitation of 20 pg, a 1/x weighting was applied to the calibration data. Accuracy and precision variation for intra and inter-day validation were below the acceptable limit (15%) for pharmacokinetic studies. PMID- 15792248 TI - Determination of organic acids in aerosol particles from a coniferous forest by liquid chromatography-mass spectrometry. AB - LC-MS methods with use of ion-trap and time-of-flight mass spectrometers were developed for the determination of organic acids in aerosol samples collected by a high-volume sampler in a Finnish coniferous forest. Comparison was made of the composition of samples collected during atmospheric formation of new aerosol particles and on days when this formation did not occur. A dynamic sonication assisted solvent extraction system was developed for fast and quantitative extraction of the filter samples. Several organic acids, including pinonic acid, pinic acid, and homologous series of n-alkanoic acids, n-alkenoic acids, and aliphatic dicarboxylic acids, were identified. In samples collected between 08:00 and 16:00 hours the concentration of pinonic acid ranged from 0.5 ng m(-3) to 3.7 ng m(-3) and that of pinic acid from 0.2 ng m(-3) to 1.5 ng m(-3). For most of the compounds identified, the trends in concentration could be explained by the differences in temperature during collection. However, concentrations of short chain n-alkanoic acids were clearly higher on the days when new aerosol particle formation occurred. PMID- 15792249 TI - Statistical-overlap theory for elliptical zones of high aspect ratio in comprehensive two-dimensional separations. AB - The low prediction by statistical-overlap theory of the numbers of singlets and peaks in two-dimensional separations containing zones represented by either circles of small number or eccentric ellipses of any number is shown to result from use of probability expressions for unbound spaces of infinite extent. An exact theory is derived for the probability of singlet formation in a reduced two dimensional space of unit length, width, and area. The probability is a weighted sum of the probabilities of singlet formation in the interior, edge, and corner regions of the space, which depend only on saturation. The weighting factors are the fractions of area associated with each region and depend on the number of zones, the aspect ratio, the saturation, and the ellipse's spatial orientation. The average numbers of doublets, triplets, and peaks in the space are approximated by combining these results with Roach's equations describing the clustering of circles in an unbound two-dimensional space. Simulations show that theory predicts the number of singlets, doublets, triplets, and peaks, when the number of zones is 25 or more, the aspect ratio is 100 or less, and the saturation is 2 or less. The relationship is derived between the aspect ratios of ellipses in the reduced space and actual separation space. Calculations are presented for comprehensive two-dimensional gas chromatography. PMID- 15792250 TI - Simultaneous determination of noradrenaline and dopamine in Portulaca oleracea L. by capillary zone electrophoresis. AB - A simple and rapid capillary electrophoretic method was developed for the simultaneous determination of noradrenaline (NA) and dopamine (DA) in Portulaca oleracea L. The buffer solution used in this method was 40 mM tris (hydroxymethyl) aminomethane (Tris)-H3PO4 at pH 2.00 containing 15% methanol. The effects of pH value, organic modifier, and applied voltage were investigated. The linear ranges of NA and DA were 0.5-100 microg/mL (r=0.9952) and 6.25-200 microg/mL (r=0.9992), respectively. The relative standard deviations of the corrected peak area were 6.73% and 4.26%, respectively. NA and DA in Portulaca oleracea L. were simultaneous determined successfully within 5.6 min. In this way, the contents of NA and DA in different parts (stem, leaves, and seeds) of P. oleracea L. and in different extracts of leaves with different solvents (distilled water, 50% methanol, and methanol) were studied. PMID- 15792251 TI - Qualitative and quantitative analysis of active flavonoids in Flos Lonicerae by capillary zone electrophoresis coupled with solid-phase extraction. AB - Flavonoids are an important bioactive group in the commonly used herbal medicine Flos Lonicerae. A new method of capillary zone electrophoresis (CZE) coupled with solid-phase extraction (SPE) was developed for simultaneous assay of flavonoid aglycones and glycosides in Flos Lonicerae. Optimum CZE separation was achieved with a background electrolyte (BGE) solution consisting of 80 mM boric acid and 20 mM phosphate acid, adjusted to pH 8.1, with 15% acetonitrile (v/v) added, and applying a separation voltage of 28 kV. The SPE method was used for pretreating the complex matrix of botanical materials and good reproducibility was obtained when avicularin was used as internal standard. Linearity of the method was excellent with correlation coefficients (r2) in the range of 0.9995-0.9999 and detection limits were lower than 0.6 microg/mL for the four flavonoids. The obtained recoveries varied between 93 to 104% while the relative standard deviations (RSDs) were below 4.4% (n=3). The developed CZE method was successfully used for the separation of eight flavonoids and the quantification of the four flavonoids in five species of Flos Lonicerae. PMID- 15792252 TI - Size fractionation of marine sediments by pinched inlet gravitational split-flow thin fractionation and the study of size dependent PCDD/Fs concentrations from different bay areas. AB - Pinched inlet gravitational split-flow thin fractionation (PI-GSF) has been applied to the continuous size fractionation of marine sediments in order to study the difference in sediment size distribution and the concentration of PCDD/Fs contained in different particle sizes. A PI-GSF channel, known to improve the separation efficiency by reducing the sample inlet thickness, was utilized to fractionate sediments collected from three different bay areas (Geoje, Ulsan, and Pohang) in Korea into 5 different sub-populations (<2.0, 2.0-5.0, 5.0-10, 10-20, 20-63 microm in diameter). The sorted sediment fractions from PI-GSF were examined using electron microscopy to obtain size distribution and the results showed a variation in particle size distribution between bay areas. When the collected particle fractions were examined for size dependent levels of PCDD/Fs, the concentrations of total PCDD/Fs were shown to be much greater for samples collected close to heavy industry complexes than sediments from bay areas without major industry. PMID- 15792253 TI - The full evaporation technique: a promising alternative for residual solvents analysis in solid samples. AB - The static headspace technique is the most common approach to residual solvent analysis in pharmaceutical and environmental matrices. This paper presents an alternative tool where the volatile impurities are released from the matrix by working directly on a small amount of sample at a high equilibration temperature: the so-called Full Evaporation Technique (FET). The capability of this method was evaluated on a mixture of nine solvents, which belong to Class 3 in the classification of the European Pharmacopoeia Agency for residual solvents, at various levels of concentration: ethanol, acetone, 2-propanol, methyl acetate, 2 butanone, ethyl acetate, tetrahydrofuran, 2-methyl-1-propanol, 1-butanol. Data on linearity, accuracy, precision and sensitivity are reported. Use of an internal standard proved to be necessary when using such a method. The method is then successfully applied to the analysis of solvent traces in permethylated beta cyclodextrin. PMID- 15792254 TI - Optimization of microwave-assisted extraction for alizarin and purpurin in Rubiaceae plants and its comparison with conventional extraction methods. AB - A simple and rapid microwave-assisted extraction (MAE) procedure was developed and optimized for two common color pigments, alizarin and purpurin, in various samples of Rubiaceae plants. Several variables that can potentially affect the extraction efficiency, namely temperature, methanol concentration in the extractant mixture, time, and solvent volume were optimized by means of a central composite design approach. The results suggest that temperature and methanol concentration in the solvent mixture are statistically the most significant factors. The separation and quantitative determination of the pigments was carried out in less than 6 min by a developed high-performance liquid chromatographic method with UV detection at 250 nm. Under optimum operating conditions, MAE showed significantly higher recoveries than those obtained by the conventional extraction methods (ultrasonic and reflux extraction), ranging from 84 to 94%. In addition, a drastic reduction of the extraction time (20 min versus 6 h) and solvent consumption (20 versus 100 mL) was achieved with a reproducibility (RSDs < 10%) comparable with that provided by the reflux extraction as a reference method. PMID- 15792255 TI - Aide memoire--the role of the German medical establishment in the Holocaust: a retrospective on the 60th Anniversary of the Liberation of Auschwitz. PMID- 15792256 TI - High frequency of pre-diabetes, undiagnosed diabetes and metabolic syndrome among overweight Arabs in Israel. AB - BACKGROUND: Increased insulin resistance, which is associated with obesity, is believed to underlie the development of metabolic syndrome. It is also known to increase the risk for the development of glucose intolerance and type 2 diabetes. Both conditions are recognized as causing a high rate of cardiovascular morbidity and mortality. OBJECTIVES: To assess the prevalence of metabolic syndrome and different glucose intolerance states in healthy, overweight Arab individuals attending a primary healthcare clinic in Israel. METHODS: We randomly recruited 95 subjects attending a primary healthcare clinic who were healthy, overweight (body mass index > 27) and above the age of 40. Medical and family history was obtained and anthropometric parameters were measured. Blood chemistry and oral glucose tolerance test were performed after overnight fasting. RESULTS: Twenty seven percent of the subjects tested had undiagnosed type 2 diabetes according to WHO criteria, 42% had impaired fasting glucose and/or impaired glucose tolerance and only 31% had a normal OGTT. Metabolic syndrome was found in 48% according to criteria of the U.S. National Cholesterol Education Program, with direct correlation of this condition with BMI and insulin resistance calculated by homeostasis model assessment. Subjects with metabolic syndrome had a higher risk for abnormality in glucose metabolism, and the more metabolic syndrome components the subject had the higher was the risk for abnormal glucose metabolism. Metabolic syndrome predicted the result of OGTT with 0.67 sensitivity and 0.78 specificity. When combined with IFG, sensitivity was 0.83 and specificity 0.86 for predicting the OGTT result. CONCLUSIONS: According to our initial evaluation approximately 70% of the overweight Arab population in Israel has either metabolic syndrome or abnormal glucose metabolism, indicating that they are at high risk to develop type 2 diabetes and cardiovascular morbidity and mortality. This population is likely to benefit from an intervention program. PMID- 15792257 TI - Susceptibility of methicillin-resistant Staphylococcus aureus to vancomycin, teicoplanin, linezolid, pristinamycin and other antibiotics. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus is a major nosocomial pathogen worldwide. Vancomycin is the traditional drug of choice, but decreasing susceptibility to vancomycin and other glycopeptides has been reported since 1996. OBJECTIVES: To test the in vitro activity of linezolid (oxazolidinone) and other antimicrobial agents against MRSA isolates recovered from hospitalized patients. METHODS: We tested 150 MRSA isolates recovered from hospitalized patients. The minimal inhibitory concentration of vancomycin, teicoplanin, pristinamycin (quinupristin-dalforistin) and linezolid was determined by the Etest method. Susceptibility to other antibiotics was tested by the disk diffusion method. RESULTS: All isolates were sensitive to vancomycin, teicoplanin, pristinamycin, and linezolid. The MIC90 was 2.0 microg/ml for vancomycin and teicoplanin (range 0.5-2.0 microg/ml and 0.125-2.0 microg/ml, respectively), and 0.5 microg/ml for pristinamycin and linezolid (range 0.125 0.75 microg/ml and 0.125-0.5 microg/ml, respectively). Of the other antibiotics, fusidic acid showed the best in vitro activity, with 96.7% susceptibility, associated with trimethoprim/sulfamethoxazole (85.8%) and minocycline (84%). Penicillin was associated with the lowest susceptibility (1.3%), associated with ofloxacin (3%) and erythromycin (14%). An increase in the minimal inhibitory concentration value of vancomycin was associated with a significant decrease in resistance to TMP-SMZ (P < 0.01) and an apparent increase in resistance to other antibiotics. CONCLUSION: The excellent in vitro activity of linezolid and its reported in vivo effectiveness renders it an important therapeutic alternative to vancomycin in the treatment of MRSA infection. PMID- 15792259 TI - Intrapartum prophylaxis of group B streptococcal disease in Israel: guidelines and practice. AB - BACKGROUND: The incidence of perinatal, early-onset Group B streptococcal sepsis is very low in Israel and there are no local guidelines for prevention of the disease. OBJECTIVES: To determine to what extent the current Centers for Disease Control guidelines are practiced in Israel, the reasons for their adoption or rejection, and the need for local official guidelines. METHODS: A telephone questionnaire was conducted of all 27 delivery units in Israel. Answers were obtained from 26, either from the clinical director or the senior obstetrician in charge at the time of the interview. RESULTS: Only in 2 of the 26 delivery units (8%) are the CDC guidelines adhered to exactly; in 6 units they are deliberately rejected, and in 8 units they are not practiced although they are allegedly implemented. Thus, the CDC guidelines are not practiced in 14 delivery units (54%). Medico-legal consideration is the sole or major reason for adopting these guidelines in 80% (16/20) of the delivery units where they are seemingly implemented. In the majority of these units (18/20) there is readiness to abandon current practice, should local guidelines differ from those of the CDC, provided that local guidelines are issued by an authoritative source. CONCLUSION: CDC guidelines are either deliberately rejected or incorrectly practiced in most Israeli delivery units. The medico-legal argument is one of the main reasons for practicing these guidelines. Since the CDC guidelines probably do not apply in Israel, official local guidelines are urgently needed. PMID- 15792258 TI - Ventilatory support during training improves training benefit in severe chronic airway obstruction. AB - BACKGROUND: One mechanism that may limit training effect in chronic obstructive pulmonary disease is the ventilatory limitation and associated dyspnea. OBJECTIVES: To minimize ventilatory limitation during training of patients with severe COPD by applying bi-level positive pressure ventilation during training in order to augment training intensity (and effect). METHODS: The study group comprised 19 patients (18 males, 1 female) with a mean age of 64 +/- 9 years. Mean forced expiratory volume in 1 second was 32 +/- 4% of predicted, and all were ventilatory-limited (exercise breathing reserve 3 +/- 9 L/min, normal > 15 L/min). The patients were randomized: 9 were assigned to training with BiPAP and 10 to standard training. All were trained on a treadmill for 2 months, twice a week, 45 minutes each time, at maximal tolerated load. Incremental maximal unsupported exercise test was performed before and at the end of the training period. RESULTS: BiPAP resulted in an increment of 94 +/- 53% in training speed during these 2 months, as compared to 41 +/- 19% increment in the control group (P < 0.005). Training with BiPAP yielded an average increase in maximal oxygen uptake of 23 +/- 16% (P < 0.005), anaerobic threshold of 11 +/- 12% (P < 0.05) and peak O2 pulse of 20 +/- 19% (P < 0.05), while peak exercise lactate concentration was not higher after training. Interestingly, in the BiPAP group, peak exercise ventilation was also 17 +/- 20% higher after training (P < 0.05). Furthermore, contrary to our expectation, at any given work rate, ventilation (and tidal volume) in the BiPAP group was higher in the post-training test as compared to the pre-training test, and the end tidal partial pressure of CO2 at 55 watts was lower, 40 +/- 4 and 38 +/- 4 mmHg respectively (P < 0.05). No improvement in exercise capacity was observed after this short training period in the control group. CONCLUSION: Pressure-supported ventilation during training is feasible in patients with severe COPD and it augments the training effect. The improved exercise tolerance was associated with higher ventilatory response and therefore lower P(ET)CO2 at equal work rates after training. PMID- 15792260 TI - Coccygectomy for intractable coccygodynia. AB - BACKGROUND: Coccygectomy is an uncommon procedure that many surgeons are reluctant to perform due to its proximity to the anus and the risk of rectal perforation and infection. OBJECTIVES: To evaluate the diagnostic accuracy and outcome of coccygectomy. METHODS: We retrospectively reviewed the operative results in nine patients (seven females and two males) who underwent coccygectomy for coccygodynia in the last 5 years following failure of conservative treatment. RESULTS: The outcome of the procedure was excellent in five patients, good in one patient and poor in two patients. CONCLUSIONS: It is mandatory to perform bone scanning in every patient with coccygodynia and before coccygectomy in order to rule out the presence of malignancy. Coccygectomy is recommended for patients with isolated coccygodynia. PMID- 15792261 TI - Torsade de pointes in patients on chronic amiodarone treatment: contributing factors and drug interactions. AB - BACKGROUND: Torsade de pointes is rarely associated with chronic amiodarone treatment, despite the effect of amiodarone on QT interval prolongation. OBJECTIVE: To identify risk factors and associated conditions that may cause TdP in patients on chronic amiodarone treatment. METHODS: We reviewed the data of six consecutive patients on chronic amiodarone treatment who were admitted to the intensive cardiac care unit due to syncope and TdP. RESULTS: The patients' median age was 73.5 years, and five were women. Concomitantly, loratadine was given to two patients and trazodone to one patient. Associated and attributing conditions to the development of TdP were hypokalemia in three patients, drug-induced bradycardia in one and reduced left ventricular function in four. CONCLUSIONS: TdP associated with chronic amiodarone treatment may occur when amiodarone is co administered with drugs that may potentially prolong QT interval. Additional risk factors for amiodarone-associated TdP include female gender, hypokalemia, reduced left ventricular function and bradycardia. PMID- 15792262 TI - Initial experience with a mechanical ventilation weaning unit. AB - BACKGROUND: While increasing numbers of patients require prolonged mechanical ventilation, resources for weaning are either limited (ICU beds) or inadequate (general wards). OBJECTIVES: To report on our initial experience over a 7 month period with an eight-bed mechanical ventilation weaning unit. METHODS: Sixty-nine patients requiring MV for > 10 days were admitted to the unit (nurse:patient ratio 1:4). Data collected included reason for MV, duration of hospital stay, and MVWU course. Outcome results (successful weaning and mortality) were compared to those in historic controls (patients ventilated in the general wards over a 4 month period prior to the MVWU; n = 100). RESULTS: The mean age of the patients was 68 +/- 16.6 years and hospital stay prior to MVWU admission 28.6 +/- 24.2 days (range 10-72). The main reasons for MV included acute exacerbation of chronic obstructive pulmonary disease (31%) and recent pneumonia (28%). Mean MVWU stay was 13.5 +/- 15.7 days (range 1-72 days). Thirty-four patients (49%) underwent tracheostomy. Fourteen patients required admission to the ICU due to deterioration in their status. Twenty-nine patients (42%) were successfully weaned and discharged to the wards. A further 20 patients were transferred to the chronic ventilation unit of a regional geriatric rehabilitation hospital, where 5 were subsequently weaned and 15 required prolonged ventilation. Compared to controls (matched for age and reason for mechanical ventilation), more MVWU patients underwent successful weaning (49% vs. 12%, P < 0.001) and their mortality rate (n = 12) was significantly lower (17% vs. 88%, P < 0.001). CONCLUSION: The higher level of care possible in a MVWU may result in a significantly improved rate of weaning and lower mortality. The assessment of long-term outcome in patients discharged to pulmonary rehabilitation centers requires further investigation. PMID- 15792263 TI - Donor recruitment and selection for adult-to-adult living donor liver transplantation in urgent and elective circumstances. AB - BACKGROUND: Adult-to-adult living donor liver transplantation is becoming an alternative to cadaveric transplantation in urgent and elective settings. Donor selection crucially affects donor safety and recipient outcome. OBJECTIVE: To present our algorithm of urgent and elective donor selection. METHODS: Urgent selection is expeditious and protocol-based. Elective selection permits a comprehensive process. Both include medical, psychosocial and surgical-anatomic evaluations. Liver volumes and vascular anatomy are evaluated with computerized tomographic angiography. Informed consent is obtained after painstaking explanations. Independent institutional committees review and approve all cases. RESULTS: Between July 2003 and June 2004 we evaluated 43 potential live donors for 12 potential recipients (fulminant hepatic failure, n = 5; chronic end-stage liver disease, n = 6; primary graft non-function, n = 1). Thirty-three candidates (76%) were excluded due to blood type incompatibility (n = 14, 42%), incompatible anatomy (n = 8, 24%)--including problematic volume distribution (n = 2) or vascular anatomy (n = 6)--psychosocial issues (n = 4, 12%), or medical co morbidity (n = 7, 22%). Five recipients (FHF, n = 4; chronic ESLD, n = 1) were successfully transplanted from living donors. In the acute setting, two patients (FHF, PGNF) died in the absence of an appropriate donor (cadaveric or living donor). In the elective group, one patient died of unexpected variceal bleeding and one received a cadaveric graft just before the planned living donor transplantation was performed. One candidate was transplanted overseas and two cases are scheduled. The ratio of compatibility for donation was 34% (10/29) for blood type-compatible candidates. CONCLUSIONS: Donor selection for living donor liver transplantation is a complex, labor-intensive multidisciplinary process. Most exclusions are due to blood type incompatibility or anatomic details. Psychosocial aspects of these donations warrant special attention. PMID- 15792264 TI - The role of primary care physicians in the Israel Defense Forces: a self perception study. AB - BACKGROUND: The health system of the medical corps of the Israel Defense Force is based primarily upon primary healthcare. In recent years, health management organizations have considered the primary care physician responsible for assessing the overall health needs of the patient and, accordingly, introduced the term "gatekeeper." OBJECTIVES: To describe and analyze how PCPs in the IDF view their roles as primary care providers and to characterize how they perceive the quality of the medical care that they provide. METHODS: We conducted a survey using a questionnaire that was mailed or faxed to a representative sample of PCPs. The questionnaire included demographic background, professional background, statements on self-perception issues, and ranking of roles as a PCP in the IDF. RESULTS: Statements concerning commitment to the patient were ranked higher than statements concerning commitment to the military organization. Most physicians perceive the quality of the medical care service that they provide as high; they also stated that they do not receive adequate continuous medical education. CONCLUSIONS: Our survey shows that PCPs in the IDF, like civilian family physicians, perceive their primary obligation as serving the needs of their patients but are yet to take on the full role of "gatekeepers" in the IDF's healthcare system. We conclude that the Medical Corps should implement appropriate steps to ensure that PCPs are prepared to take on a more prominent role as "gatekeepers" and providers of high quality primary medical care. PMID- 15792265 TI - The role of apoptosis in preeclampsia. AB - Altered apoptosis is involved in each step of the pathogenesis of preeclampsia. While deficient apoptosis may induce a maternal immune response against the fetus, enhanced apoptosis may interfere with the process of placentation, placental ischemia and subsequently, systemic endothelial damage. Thus, treatment modalities to inhibit or acclerate apoptosis cannot be employed in early pregnancy for prevention. Later in pregnancy, it would make sense to test treatment modalities that inhibit hypoxia-mediated apoptosis in patients in whom early testing, such as abnormal Doppler of the uterine artery, indicates the initiation of preeclampsia. This, however, has first to be tested in animal models. Since multiple different mechanisms, rather than a single factor, could contribute to the development of apoptosis, further studies to clarify the signaling mechanisms of apoptosis in preeclampsia should be conducted before any investigational treatment modalities are employed. PMID- 15792266 TI - Pharmacologic prophylaxis against nerve agent poisoning. AB - Nerve agent poisoning is characterized by the rapid progression of toxic signs, including hypersecretions, tremor, convulsions and profound brain damage. In the political arena of today's world, the threat of nerve agent use against military troops has prompted armies to search for prophylactic protection. The two main strategies for prophylaxis include biological scavengers that can bind or cleave nerve agents before they react with acetylcholinesterase, and antidotes as prophylactic treatment. Pyridostigmine is the current pretreatment for nerve agent poisoning and is in use by most of the armed forces in Western countries. However, since pyridostigmine barely crosses the blood-brain barrier it provides no protection against nerve agent-induced central injury. Pyridostigmine is ineffective when administered without post-exposure treatment adjuncts. Therefore, other directions for prophylactic treatment should be explored. These include combinations of carbamates (reversible AChE inhibitors) and central anticholinergics or NMDA receptor antagonists, benzodiazepines or partial agonists for benzodiazepine receptor, and other central AChE inhibitors approved for Alzheimer's disease. The transdermal route is an alternative way for delivering the prophylactic agent. Administration of prophylaxis can be extended also for civilian use during wartime. PMID- 15792267 TI - Neurologic aspects of neonatal hypoglycemia. AB - Profound neonatal hypoglycemia is one of the leading causes of brain injury. Hypoglycemic encephalopathy is caused by lack of glucose availability to brain cells. Although sharing a similar pathogenesis with hypoxic-ischemic encephalopathy, hypoglycemic brain insult has distinctive metabolic, brain imaging, electroencephalographic and histopathologic findings. PMID- 15792268 TI - Pita and Cola--a devastating mixture? PMID- 15792269 TI - Methicillin-resistant Staphylococcus aureus: past, present, and--too much of a- future. PMID- 15792270 TI - Training programs for patients with severe chronic obstructive pulmonary disease. PMID- 15792271 TI - Lymphomatoid granulomatosis: a diagnostic challenge. PMID- 15792272 TI - A returned traveler with dengue fever and visual impairment. PMID- 15792273 TI - Unusual presentations of infective endocarditis. PMID- 15792274 TI - Chronic lateral compression of the pelvis. PMID- 15792275 TI - Testicular adrenal rest tumors. PMID- 15792276 TI - Development of an AIDS vaccine: a daunting epidemiological challenge. PMID- 15792277 TI - Ten lessons to be learned from the withdrawal of Vioxx (rofecoxib). PMID- 15792278 TI - A comparison of response rate and time according to the survey methods used: a randomized controlled trial. AB - The purpose of this study was to evaluate the effects of recorded and standard deliveries, and stamped and franked return envelops in a postal survey of Korean Vietnam veterans. Nine hundred veterans were randomly divided into four subgroups. A randomized controlled trial was conducted for two mailing strategies. The 52 study subjects to whom mail was undeliverable and 36 additional study subjects found not to be residing at the listed addresses were excluded from the study. The 699 (86%) surveys were returned within 39 business days after the first mailing. The response rate for the recorded delivery (88%) was significantly higher than that of the standard delivery (82%)(p = 0.03), and the response rate of the stamped return envelops (88%) was higher than that of the franked return envelops (85%)(p = 0.27). The replies for the recorded and standard deliveries arrived an average of 10.2 and 9.9 business days, respectively, after the first mailing (p = 0.60). The average times of the responses for the stamped and franked return envelops were 9.8 and 10.4 business days, respectively (p = 0.25). Recorded deliveries significantly increased the response rate compared to standard deliveries, and stamped return envelops slightly increased the response rate a little compared to franked return envelops. The timing of response of recorded deliveries was similar to that of standard deliveries but the volume of response of recorded deliveries was higher than that of standard deliveries. PMID- 15792279 TI - Demands on web survey tools for epidemiological research. AB - In countries where the Internet access is high, a web-based questionnaire could save time and money compared to printed questionnaires, mainly by eliminating the two steps of transferring answers from printed to a digital data set and manually completing missing and impossible answers. However, many of the features wanted for conducting large epidemiological studies are not available in many web survey systems. Here we describe design issues the investigator needs to be aware of when using web-based questionnaires in epidemiological research. PMID- 15792280 TI - Socioeconomic deprivation and waist circumference in men and women: The Scottish MONICA surveys 1989--1995. AB - Socioeconomic deprivation and waist circumference were measured in three Scottish MONICA cross-sectional surveys of 2233 men and 2516 women aged 25-64 years in 1989-1995. Means of waist circumference, waist/hip ratio (WHR) and body mass index (BMI) increased with level of deprivation (measured by the Carstairs index) more significantly in women than in men, and more significantly in non-smokers than in current-smokers. Their obesity cases defined by conventional cut-points showed similar patterns of relation to deprivation. There appeared to be more obviously consistent and significant increases in the prevalence of large waist circumference with deprivation for both sexes than in the prevalence of WHR and BMI above the 90th centile. Also there was a more significant trend of increase in waist circumference over time than there was in WHR and BMI for both sexes. Residual case-control analysis, controlling for height, showed a 'dose-response' relationship between deprivation and waist circumference. Compared to the most affluent (the first tertile of the Carstairs score), odds ratio for men in the middle group (the second tertile) adjusted for age, survey year and smoking status was 1.37 (95%CI 1.10-1.70) and in the most deprived (the third tertile) 1.46 (1.17-1.82); and for women 1.22 (0.99-1.50) and 1.81 (1.47-2.23). The study suggests that large waist circumference, increasingly prevalent, is directly related to socioeconomic deprivation, and greater attention should be paid to increasing girth in the socially deprived. PMID- 15792281 TI - Alcohol drinking and colorectal cancer: a population-based prospective cohort study in China. AB - STUDY OBJECTIVE: To asses the association between alcohol consumption and the risk of colorectal cancer (CRC) in the Chinese population. DESIGN: A population based prospective cohort study was initiated from the colorectal cancer screening population in Jiashan County in 1989-1990. The drinking habits of individuals were investigated with demographic information. SETTING: A cohort study was followed-up from 1st May 1990 to 1st January 2001 and censored at the date of diagnosis of CRC, at death from any causes, or at 1st January 2001, whichever came first, and the person-time was computed. PARTICIPANTS: Two hundred and forty two CRC patients were diagnosed during the study period and 64,100 individuals finished the follow-up. RESULTS: The distribution of sex, smoking status, occupation, education level and marital status were all significantly different among different drinking habits at baseline. When the above factors were adjusted, no significant association was observed between alcohol consumption and the risk of CRC. Exclusion of individuals diagnosed cancer less than 1 year after the examination date did not alter the strength of an alcohol-CRC relationship. Further analysis in sex strata also did not show a significant relationship. CONCLUSIONS: Alcohol drinking may not be associated with a higher risk of CRC in the Chinese population. PMID- 15792282 TI - Moderate physical activity and prostate cancer risk: a case-control study in China. AB - A case-control study was conducted to investigate the association between physical activity and prostate cancer risk in China. A total of 130 recent histopathologically confirmed cancer cases and 274 controls were interviewed. Information collected included type, intensity and duration of various physical activities. The adjusted odds ratios of prostate cancer risk for physical activity variables were obtained from multivariate logistic regression models. Results showed that moderate physical activity was inversely related to the prostate cancer risk, with adjusted odds ratio being 0.20 (95% confidence interval 0.07-0.62) for the upper vs. the lower quartiles of weekly metabolic equivalent task-hours. The dose-response relationship was also significant (p = 0.015). The findings suggest that moderate physical activity may be protective against prostate cancer for Chinese men. PMID- 15792283 TI - The smoker's paradox and the real risk of smoking. AB - No satisfactory explanations have been offered for the smoker's paradox, the greater short-term survival of smokers after a myocardial infarction nor for the large variations in the coronary risk rate for smoking ranging between 1 and 5.9. These discrepancies as well as the smoker's paradox may be caused by different baseline characteristics of smokers and nonsmokers, whereas the usually quoted coronary risk of 2 is derived from studies based on the assumption of equal baseline characteristics. As neither this assumption nor the possibility of unequal starting conditions have been tested, we examined the main cardiovascular risk factors in smoking and nonsmoking boys as near as possible to baseline, at the age of fourteen. This age appeared to be best suited, because boys starting to smoke early are most likely to become regular and heavy smokers. Of 336 boys, 39 had smoked 8.3+/-6.0 cigarettes/day for 15.5+/-11.2 months. Compared to nonsmokers, boys who started to smoke early had lower LDL cholesterol and alpha2 antiplasmin, greater handgrip strength, vital capacity and forced expiratory volume, better perfomance on bicycle ergometry and higher testosterone. The differences in total cholesterol, LDL cholesterol, vital capacity, handgrip strength, testosterone and alpha2-antiplasmin persisted after adjustment for age, body mass, and testosterone. In addition, the differences in perfomance on bicycle ergometry and forced expiratory volume persisted after adjustment for age. These favourable baseline characteristics of those starting to smoke early can explain the smoker's paradox. In addition, they suggest that the individual coronary risk in smokers is considerably higher than 2, because the assumption of equal baseline characteristics of smokers and nonsmokers cannot be upheld. PMID- 15792284 TI - Long-term monitoring of the mortality trend of Huntington's disease in Austria. AB - Recent increasing incidence and prevalence rates of Huntington's disease (HD), a fatal neurodegenerative disorder, prompted us to investigate the epidemiological dynamic of HD in Austria during the period 1970--2001. Our study demonstrated a stable HD mortality rate throughout Austria of 0.125 per 100,000 individuals during the investigated period. The median age at death from HD was 56.5 years for both sexes and remained stable during the entire period observed, indicating no prolonged survival. An above average mortality rate from HD was observed in the north-eastern parts of Austria. Our data reflect the epidemiology of HD prior to and after the availability of genetic testing and provide a solid baseline for future investigations on the epidemiology of HD. PMID- 15792285 TI - Evaluation of non-response bias in mental health determinants and outcomes in a large sample of pre-adolescents. AB - Since non-response may jeopardize the validity of studies, comprehensive assessment of non-response is a prerequisite for proper interpretation of study findings. Recently, the baseline assessment of the TRacking Adolescents' Individual Lives Survey (TRAILS), a prospective cohort study among Dutch pre adolescents, was completed. The aim of this report is to examine non-response bias by comparing responders and non-responders regarding mental health determinants (e.g., demographics and cognitive performance) and outcomes, as well as associations between the two. Furthermore, we examine whether extended efforts to recruit participants contribute to the prevention or reduction of non-response bias. Thanks to various recruitment procedures, the initial response rate of 66% increased to a final rate of 76%. The extended efforts to recruit participants prevented non-response bias in the prevalence rates of psychopathology. Although non-responders differed from responders with respect to several individual characteristics, no significant differences were found regarding associations between these characteristics and psychopathology. We conclude that TRAILS provides a solid basis to improve our understanding of the development of mental health during adolescence. PMID- 15792287 TI - Growing recognition of the importance of service user involvement in mental health service planning and evaluation. AB - Service user involvement in the planning and provision of mental health services has been growing over the last two decades, especially in countries and areas where institutional service provision has been changed to a community-orientated model of care. However, the material involvement of service users in mental health research is still in its infancy. The aim of this paper is to attempt to place these developments in a conceptual context, to summarise the ethics-based and evidence-based reasons why it has to be considered as necessary, and to illustrate some of the emerging evidence which shows the advantages to be gained from it. In particular the results of recent studies are briefly reported, showing that outcomes data rated by service users in some cases are more important than those rated by staff. The reduction in patient-rated unmet needs in the social domain was the strongest predictor of an increase in subjective quality of life. The importance of including service user preferences within the content of the research questions is exemplified by the results of a recent study that showed that joint crisis plans can significantly reduce the use of compulsory admission during crises and by a review that demonstrated that the use of an explicit service user perspective produced distinctive insights into the long-term effects of Electro-Convulsive Therapy (ECT). PMID- 15792288 TI - Developing the theoretical basis for service user/survivor-led research and equal involvement in research. AB - AIMS AND METHODS: This article, written from a service user/survivor perspective, explores a hypothesis which seeks to offer a more systematic basis for the full and equal involvement of mental health service users/survivors in both the research process and research structures more generally. The hypothesis challenges traditional emphasis on positivist assumptions about the priority of values of 'distance', 'neutrality' and 'objectivity' (which it argues discriminate against service users and their experiential knowledge). It explores instead the idea that 'the shorter the distance between direct experience and its interpretation, then the less likely resulting knowledge is to be inaccurate, unreliable and distorted.' RESULTS AND CONCLUSIONS: The proposal discusses ways in which such (objective and subjective) distance may be reduced, to improve the quality of research, to enable more equal involvement of service users and their direct experience and to make it possible for non-service user researchers to work alongside service users on more equal terms. PMID- 15792286 TI - Exposure to ambient air pollution and prenatal and early childhood health effects. AB - Over the last years, concern for the possible influence of exposure to air pollutants in children during gestation or the first years of life has grown; exposure levels which may be reached nowadays in our dwellings and in our streets. In the present study evidence over the possible impact of ambient air pollution on the foetus and the infants (i.e.: less than 1 year) published during the last decade, 1994--2003, are revised. Studies on infant mortality and exposure to particles show an outstanding consistence in the magnitude of the effects, despite the different designs used. As a whole, data show that an increase in 10 microg/m3 of particle concentration (measured as PM10) is associated with to about 5% increase in post-neonatal mortality for all causes and around 22% for post-neonatal mortality for respiratory diseases. Regarding damage in foetal health, although results are not always consistent, most studies show associations with exposure to air pollution during pregnancy. However, the precise mechanisms of action of air pollutants on adverse reproductive results are still unknown, so is the period of exposure most relevant during pregnancy and the specific pollutant which may represent a higher risk. Follow-up studies evaluating personal exposure to different air pollutants are required, allowing for the adequate evaluation of the impact of each pollutant in different periods of pregnancy, as well as providing hypotheses on their possible mechanisms of action. PMID- 15792289 TI - User/consumer involvement in mental health service delivery. AB - The involvement of mental health service users in service delivery is a new and growing phenomenon. Such involvement is complex, given the history of paternalism in the mental health system, the power differential between service providers and service users, and the very differing views each group holds on multiple issues. Unless such differences are addressed, there can be no meaningful involvement. Service user involvement needs to apply to all aspects of the service delivery system, including professional training, service design, delivery, evaluation, and research. User/survivors, and their organizations, have developed a body of experience and knowledge that needs to be recognized and respected. Unless there are multiple opportunities for ongoing and open dialogue on these many difficult issues, real user involvement will not occur. PMID- 15792290 TI - All about PSY DREAM. Psychiatric Drug Registration, Evaluation & All-Inclusive Monitoring. AB - AIMS: In this article the author--board-member of the European Network of (ex-) Users and Survivors of Psychiatry (ENUSP)--explains, what is needed to guarantee a minimal level of involvement of users and survivors of psychiatry into issues relating to psychiatric drugs. METHODS: He reflects demands of their organisations, and compares these demands with the current involvement level. Considering the concrete circumstances in psychiatry, he reflects the risks and dangers of the administered drugs--especially the widely used neuroleptics--for example enhanced breast cancer risk in women, suicidal effects, receptor changes, tardive dyskinesia and other toxic reactions. RESULTS: Considering the unique situation of psychiatric patients to receive treatment in general without informed consent and in a violent way or through bullying and threat, he argues for to provide their involvement in all aspects of psychiatric drug issues- especially registration and monitoring, for example by their involvement in creating and running a suicide register. And he argues for involvement in ethics committees, licensing processes and providing guidelines and decision making about effectiveness and reimbursement of costs. CONCLUSIONS: As first steps towards these aims he proposes independent and user-controlled research, independent and user-controlled education and independent and user-controlled information about the effects of psychiatric drugs. PMID- 15792291 TI - Attuned practice: a service user study of specialist child and adolescent mental health, UK. AB - AIMS: Best practice emphasises user involvement. This exploratory study addresses the views of teenage clients and their parents on service delivery in a specialist Child and Adolescent Mental Health Service (CAMHS) serving a population of 250,000. It aims to explore some of the complexities inherent in children's services when parents are integral to modes of treatment. METHODS: Twenty-seven teenage clients from specialist CAMHS were recruited with their parents (n=30). All were white British, 11 boys and 16 girls, from a range of socioeconomic backgrounds. Focus groups were employed using a series of structured interactive technique to elicit information, preceded by home visits. Analysis of interview data followed standard approaches to qualitative data analysis. Descriptive statistics were generated from both home interview data and focus groups. RESULTS: Three themes emerged: the core values implicated in establishing a therapeutic alliance; the style of therapy and mode of practice (i.e. its inclusiveness of different family members). PRACTICE IMPLICATIONS: Core therapeutic skills are of fundamental importance. Our paper supplements a model of organisational user involvement with a model of therapeutic user involvement for use in negotiating mode of practice. CONCLUSIONS: This exploratory study was a collaboration between service users, researchers and health professionals exploring three important themes of therapy and the complexities inherent in children's services. The process of eliciting views was therapeutic in itself leading to the formation of a parent-led self-help group. The design can be replicated in other specialist CAMHS to achieve attuned practice. PMID- 15792292 TI - Deliberate self-injury. A consumer-therapist co-run group. A choice or a necessity? AB - AIMS: This aim of this paper is to discuss a consumer-therapist co-run pilot group on self-injury held at an American college. Deliberate self-injury has come to be a common phenomenon in schools and colleges. However the treatment and understanding of self-injury still remains a challenge. Most people who self injure remain hidden in society and do not seek therapy, due to the shame-filled stereotypes and misconceptions that surround self-injury. Group therapy has been discussed as ineffective by many in the mental health field while individual treatment has been controversial for decades. METHOD: The paper will discuss how this group treatment was informed by consumer empowerment philosophy and how consumer and therapist facilitators worked together to turn it into a healing experience. RESULTS: This group was indeed challenging but confirmed that consumer-run programs are able to function as a catalyst to facilitate healing. It confirmed that breaking away from traditional models that allocate all power and hope for change to professionals might not only be an important step but, in cases such as treatment of self-injury, a necessity. PMID- 15792293 TI - Who's mind is it anyway? AB - As UK Health Minister John Bowis knew the challenges, the progress and the disasters of mental health provision and policy. He learned from service users, carers and practitioners that reality was usually different from theory. He came to believe a new structure was necessary to give patients a one-stop shop of care and treatment, that patients should be more involved in decisions that affect them and that stigma must be tackled by education and by wider understanding of services and crisis management. He now campaigns in the European Parliament and in many parts of the world for mental health to be higher on national agendas and to be based on effective and humane policies that ensure the appropriate treatment is provided and provided with dignity. PMID- 15792294 TI - Mental health services in Central and Eastern Europe: current state and continuing concerns. AB - AIMS: This paper attempts to examine the current state of mental health services in Central and Eastern Europe. METHODS: Selected review of available literature has been done with emphasis on information complied by World Health Organization (WHO). RESULTS: The magnitude and burden of mental disorders is high in Europe, the mental health services are inadequate in most Central and Eastern European countries and human rights situation is unsatisfactory. However, there are some positive recent developments worth noting. These include increased attention to human rights, cooperation and collaboration at sub-regional level and emergence of family and consumer associations. CONCLUSIONS: A concerted and systematic attempt needs to be made to respond to the challenge of providing adequate and human rights-based mental health services in Central and Eastern European countries. The framework developed by WHO can assist the countries in their initiatives aimed at improving mental health systems. PMID- 15792295 TI - Summary health statistics for U.S. children: National Health Interview Survey, 2003. AB - OBJECTIVES: This report presents both age-adjusted and unadjusted statistics from the 2003 National Health Interview Survey (NHIS) on selected health measures for children under 18 years of age, classified by sex, age, race, Hispanic origin, family structure, parent's education, family income, poverty status, health insurance coverage, place of residence, region, and current health status. The topics covered are asthma, allergies, learning disability, Attention Deficit Hyperactivity Disorder (ADHD), use of prescription medication, respondent assessed health status, school-loss days, usual place of health care, time since last contact with a health care professional, unmet dental needs, time since last dental contact, and selected measures of health care access. SOURCE OF DATA: The NHIS is a multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics and is representative of the civilian noninstitutionalized population of the United States. Data are collected for all family members during face-to-face interviews with adults present at the time of interview. Additional information about children is collected for one randomly selected child per family in face-to-face interviews with an adult proxy respondent familiar with the child's health. SELECTED HIGHLIGHTS: In 2003, most U.S. children under 18 years of age had excellent or very good health (83%). However, 10% of children had no health insurance coverage, and 5% of children had no usual place of health care. Thirteen percent of children had ever been diagnosed with asthma. An estimated 8% of children 3-17 years of age had a learning disability, and an estimated 6% of children had ADHD. PMID- 15792296 TI - Adsorption of arsenate and arsenite by iron-treated activated carbon and zeolites: effects of pH, temperature, and ionic strength. AB - Naturally occurring arsenic contaminates groundwater in many countries, including the United States, at levels greater than 10 microg l(-1), the current WHO guideline value, increasing the risk of skin, lung, bladder, and kidney cancer in millions of people. Arsenic toxicity is dependent on its chemical form; arsenite is more toxic due to its higher affinity for protein than arsenate. This study supports worldwide research efforts to obtain drinking water with arsenic levels below 10 microg l(-1). Batch adsorption kinetic and isotherm studies were conducted to compare and evaluate iron-treated adsorbents for arsenate and arsenite removal from aqueous media. Two iron treatments were investigated as well as the effects of varied pH, temperature, and ionic strength increases on adsorption effectiveness. Adsorbent materials such as activated carbon and naturally occurring zeolites (clinoptilolite and chabazite) were selected because of their relative low cost and because the zeolites are potential point-of-use materials for mitigating arsenic contaminated groundwater. Molecular sieves, Faujasite (13X) and Linde type A (5A) were selected because they provide a basis for comparison with previous studies and represent well-characterized materials. Iron-treated activated carbon and chabazite showed the most promise as low-cost arsenic adsorbents; activated carbon removed approximately 60% of arsenate and arsenite while chabazite removed approximately 50% of arsenate and 30% of arsenite. Modeling arsenate and arsenite adsorption by these adsorbents using the Langmuir and Freundlich isotherm expressions determined the adsorbents' capacity for arsenic removal from aqueous media. Arsenate removal by iron-treated activated carbon and clinoptilolite best fit the Langmuir model. Arsenate removal by iron-treated chabazite and arsenite removal by activated carbon, chabazite, and clinoptilolite best fit the Freundlich model. Applications of iron-modified activated carbon for effective arsenate removal would require pH values between 7 and 11, chabazite between 4 and 5, and clinoptilolite between 3 and 7. Arsenite removal by iron-modified activated carbon would require pH values between 7 and 11, chabazite between 7 and 10, and clinoptilolite between 4 and 11. Increasing temperature improved adsorption performance for activated carbon and the zeolites. Increasing ionic strength improved performance of iron-treated activated carbon and zeolites. PMID- 15792297 TI - Advanced oxidation degradation of dichlorobenzene in water by the UV/H2O2 process. AB - Through UV/H2O2 photocatalytic advanced oxidation technique the performance, kinetics, pathway, and mechanism of ortho-dichlorobenzene (O-DCB) in water and the effect parameters of degradating reaction were studied systematically. The results showed that the reaction of photocatalytic degradation of O-DCB was fast and accorded well with the pseudo-first-order kinetics. The roles of some parameters, such as the pH, the initial concentrations of O-DCB, H2O2, and some anions in reaction solution, were examined in detail. It was found that weak acidic or neutral environment was favorable to the degradation reaction and some anions slowed down the photocatalytic degradation rate. Input amount of H2O2 possessed the best value under specified condition. The intermediates of O-DCB degradation were also tentatively identified 2,3-dichlorophenol, 3,4 dichlorophenol, formic acid, acetic acid, and oxalic acid mainly adopting GC/MS and ion chromatogram (IC) techniques. According to this, the pathway and mechanism of UV/H2O2 photocatalytic degradation of O-DCB were deduced and led. PMID- 15792298 TI - Removal of earthy-musty odorants in drinking water by powdered activated carbon. AB - The removal of four earthy-musty compounds, 2-methylisoborneol (MIB), 2,4,6 trichloroanisole (TCA), 2-isopropyl-3-methoxy-pyrazine (IPMP), and 2-isobutyl-3 methoxy-pyrazine (IBMP), by powdered activated carbon (PAC) were investigated under the simulative condition of Xicun water plant, Guangzhou. The adsorption kinetics of odor compounds by two PACs show that main removal of odor compounds occurs within contact time of 1 h. Compared with the wood-based PAC, the coat based PAC evidently improves the removal efficiency of poorly adsorbed compounds like MIB. The effects on removal efficiency such as optimum PAC dosage, initial concentration of the organics, chlorine residual, background organics, and changes of water quality were investigated. The percent removal of trace odorants at any given time for a particular carbon dosage is irrelative to the initial concentration of the odor compounds. Adsorptive capacity of PAC for target compounds is reduced by chlorine residual and background organics. Characteristics of raw water have vast influence on the removal of target compounds by PAC. PMID- 15792299 TI - Indicator organisms associated with stormwater suspended particles and estuarine sediment. AB - This study examined the fate of indicator organisms (fecal coliform, E. coli, and enterococci) from urban stormwater runoff associated with estuarine sediments and stormwater suspended particles. The research found a significant increase of fecal coliform, E. coli, and enterococci in the estuarine sediments at study sites following a given stormwater event. The sedimentation mechanism was linked to an increase in indicator organisms in the estuarine sediments. These indicator organisms attached to stormwater suspended particles, which then settled to the bottom sediment. A higher percentage of attachment onto stormwater suspended particles was observed for E. coli as compared to enterococci and fecal coliform. Among the three organisms, enterococci preferentially attached to the particles with diameters from 10 microm to 30 microm, while fecal coliform and E. coli had a broader distribution. Estuarine sediments were found to prolong survival of indicator organisms for at least seven days prior to reduction to background levels. PMID- 15792300 TI - Kinetics of wet air oxidation (WAO) of alcaloide factory wastewater. AB - Wet air oxidation (WAO) of Afyon alcaloide factory wastewater, a typical high strength industrial wastewater, was carried out. The process was performed in a specifically designed titanium bubble reactor at temperatures in the range of 140 160 degrees C. The kinetics of WAO of alcaloide factory wastewater was modeled by assuming two distinct steps. The rates of destruction were measured with respect to reduction in COD. The oxidation reaction was found to be first order with respect to COD concentration and also second order with respect to oxygen concentration in both steps. The values of activation energies were found to be in the range of 4.93 x 10(4)-7.85 x 10(4) kJ/kmol. PMID- 15792301 TI - Adsorption-desorption characteristics of cadmium in variable charge soils. AB - Cadmium (Cd) has received considerable attention because of its association with various human health problems. The behavior of adsorption-desorption of Cd at contaminated levels in two variable charge soils were investigated. The red soil (RAR) developed on the Arenaceous rock (clayey, mixed siliceous thermic typic Dystrochrept) adsorbed more Cd2+ than the red soil (REQ) derived from the Quaternary red earths (clayey, kaolinitic thermic plinthite Aquult). The characteristics of Cd adsorption could be described by the Freundlich equation (r2 = 0.997 and 0.989, respectively, for the RAR and REQ) and the simple Langmuir adsorption equation (r2 = 0.985 and 0.977, respectively, for the RAR and REQ). The maximum adsorption values (Xm) that were obtained from the simple Langmuir model were 36.23 mmol Cd2+ kg(-1) soil and 31.15 mmol Cd2+ kg(-1) soil, respectively for the RAR and REQ. Adsorption of Cd2+ decreased soil pH by 1.28 unit for the RAR soil and 1.23 unit for the REQ soil at the highest loading. The distribution coefficient (kd) of Cd in the soil decreased exponentially with increasing Cd2+ loading. The adsorption of cadmium in the two variable charge soils was characterized by a rapid process that lasted approximately 15 min, followed by a slower but longer period. 85.5% and 79.4% of the added Cd were adsorbed within two hours by the RAR and REQ soil, respectively. More Cd2+ was adsorbed at 10 degrees C than at 25 degrees C or 40 degrees C. After five successive desorptions with 0.01 mol L(-1) NaNO3 solution, 53.3% of the total adsorbed Cd2+ in the RAR soil was desorbed and the corresponding value of the REQ soil was 46.5%, indicating that the RAR soil had a lower affinity for Cd2+ than the REQ soil at the same Cd2+ loading. PMID- 15792303 TI - Copper phytoavailability and uptake by Elsholtzia splendens from contaminated soil as affected by soil amendments. AB - Pot and field experiments were conducted to evaluate bioavailability of Cu in contaminated paddy soil (PS) and phytoremediation potential by Elsholtzia splendens as affected by soil amendments. The results from pot experiment showed that organic manure (M) applied to the PS not only remarkably raised the H2O exchangeable Cu, which were mainly due to the increased exchangeable and organic fractions of Cu in the PS by M, but also stimulated plant growth and Cu accumulation in E. splendens. At M application rate of 5.0%, shoot Cu concentration in the plant increased by four times grown on the PS, so as to the elevated shoot Cu accumulation by three times as compared to the control. In the field trial, soil amendments by M and furnace slag (F), and soil preparations like soil capping (S) and soil discing (D) were performed in the PS. Soil capping and discing considerably declined total Cu in the PS. Application of M solely or together with F enhanced plant growth and increased H2O exchangeable Cu levels in the soil. The increased extractability of Cu in the rhizosphere of E. splendens was noted, which may have mainly attributed to the rhizospheric acidification and chelation by dissolved organic matter (DOM), thus resulting in elevating Cu uptake and accumulation by E. splendens. Amendments with organic manure plus furnace slag (MF) to the PS caused the highest exactable Cu with saturated H2O in the rhizospheric soil of E. splendens after they were grown for 170 days in the PS, thus achieving 1.74 kg Cu ha(-1) removal from the contaminated soil by the whole plant of E. splendens at one season, which is higher than those of the other soil treatments. The results indicated that application of organic manure at a proper rate could enhance Cu bioavailability and increase effectiveness of Cu phytoextraction from the contaminated soil by the metal-tolerant and accumulating plant species (E. splendens). PMID- 15792302 TI - Phytoextraction of Zn and Cu from sewage sludge and impact on agronomic characteristics. AB - The presence of elevated concentrations of heavy metals limits the usage of sewage sludge as a fertilizer and soil amendment. Experiments were carried out to examine the extent to which seven plant species phytoextracted Zn and Cu from dewatered sludge. The hyperaccumulators Thlaspi caerulescens and Sedum alfredii showed the greatest removal of Zn, while shoots and tubers of two species of Alocasia showed the greatest Cu removal. Cultivation of plants in the sludge resulted in significant decreases in total Zn and changes in the partitioning of Zn between soil pools. However, Cu levels were largely unchanged and remained associated predominantly with the organic matter pool. Agronomic characteristics of the sludge material, such as pH, organic matter content, and nitrogen, phosphorus, and potassium concentrations, did not change significantly during the four-month growth period, indicating that subsequent crops could be sustained by this material. These results suggest that Zn can be phytoextracted from sludge material, provided the rate of metal uptake exceeds the rate of mobilization to the exchangeable fraction. Since there was no appreciable accumulation of Zn and Cu in seeds of Zea mays in this study, some tissues from sludge-grown plants could potentially be used as animal fodder. PMID- 15792304 TI - Biosorption of rare earth metal ion on aerobic granules. AB - Aerobic granules are microbial aggregates possessing excellent settling ability and high-porosity structure. In this study, aerobic granules as a novel type of biosorbent were used for cerium's removal from aqueous solution simulating the polluted industrial wastewater. Batch trials were conducted at different initial cerium ion and granule concentration. Biosorption kinetics was also studied. The biosorption conformed to a first-order kinetics model. The results showed that the biosorption ability of aerobic granules was related to both initial cerium ion and granule concentration. The maximum biosorption capacity of cerium by aerobic granules was 357 mg g(-1) granules. The aerobic granules were settled down by gravity from the aqueous solution in one minute after the biosorption experiments. Thus, the post-separation of the conventional suspended biosorbents from the treated effluent could be ignored in the aerobic granule-based biosorption process. All the results confirmed the technical feasibility of the biosorption process by aerobic granules. PMID- 15792305 TI - Evaluation of toxic properties of industrial wastewater using on-line respirometry. AB - An on-line respirometric biosensor using activated sludge microorganisms was used in this work for the assessment of toxic properties of industrial effluents and for the evaluation of their effect on the performance of a municipal wastewater treatment plant. Continuous monitoring of combined industrial discharges was carried out for a period of six months, and increased toxicity values were observed during the evening times and the weekends, possibly due to the discharge of partially treated wastewaters from certain processing units. Furthermore, no direct correlation was observed between toxicity values and organic content, indicating that simply monitoring organic load, as it usually happens in most wastewater treatment plants, does not help in predicting the toxic properties of the effluent. Therefore, the installation and operation of an on-line respirometer is required, in combination with chemical analysis, for the protection of the operation of a municipal wastewater treatment plant from toxic shocks. The assessment of toxicity of wastewaters from certain industrial plants showed that bioluminescence inhibition on Vibrio fischeri was generally higher than the respirometric inhibition on activated sludge microorganisms. It was concluded that for the evaluation of potential toxicity of influents on a wastewater treatment plant, activated sludge microorganisms should be preferred as test species. PMID- 15792306 TI - A laboratory and pilot study of thermally enhanced soil vapor extraction method for the removal of semi-volatile organic contaminants. AB - Laboratory and field pilot study was conducted to evaluate the effectiveness of thermally enhanced soil vapor extraction (SVE) system for the removal of semi volatile organic contaminants (SVOCs) from soils. Several parametric studies were conducted to evaluate the effect of temperature and air flow rate on the removal of SVOCs. Column studies near ambient temperature show very little efficiency for SVOCs removal. The removal rates of SVOCs are highly dependent on temperature. Moving fronts in the effluent concentrations of contaminants are observed, according to the order of volatility of contaminant. This result is attributed to continuous changes in the liquid composition toward less volatile fractions, thus increasing the mole fraction of the less volatile fractions remaining in the liquid mixture. It is also found that increased air flow rate results in high removal rate of contaminants. However, too high air flow rate brings about the mass transfer limitation on the volatilization of the contaminants. Results from steady-state column studies show that mass transfer limitation is reduced at high temperature, possibly due to the increase in the diffusivity of contaminant in the liquid and gas phase. A field pilot study of a hot air injection method for the remediation of diesel-contaminated soil has shown advantages over the conventional SVE. Within 30 days of test operation, TPH concentrations at unsaturated soils were dramatically reduced, corresponding to more than 95% removal. However, temperature profiles and the removal rates of SVOCs near groundwater level indicate that moisture content would be a limiting factor in the real application of hot air injection. PMID- 15792307 TI - Concentration of selected metals in muscle of various fish species. AB - The purpose of this study was to assess concentration of selected metals (Fe, Mn, Zn, Cu, Ni, Co, Cr, Pb, Cd, and Hg) in the muscle of four common Slovak fish species (chub--Leuciscus cephalus, barbel--Barbus barbus, roach--Rutilus rutilus, and perch--Perca fluviatilis). Furthermore, correlations among the selected metals and order of metal accumulation in the fish muscle were determined. An electrofishing technique was used for collecting the fish from the Nitra River (Slovakia) in September 2003. Concentrations of selected metals were measured using an atomic absorption spectrophotometer Pye Unicam SP9. The concentrations of metals (mg/kg wet weight basis) ranged as follows: Fe 3.41-15.14; Mn 0.20 0.81; Zn 3.51-15.64; Cu 0.25-0.78; Ni 0.07-0.25; Co 0.05-0.19; Cr 0.11-0.42; Pb 0.20-5.81; Cd 0.06-0.56, and Hg 1.35-6.52. Significant correlations (P < 0.05) between Fe-Cu, Fe-Ni, Fe-Cr, Mn-Ni, Mn-Cr, Mn-Cr, Cu-Ni, and Ni-Cr were observed. The level of lead exceeded the maximum allowable concentration in Slovakia by Codex Alimentarius for safe human consumption (0.2 mg/kg) in the majority of samples (97.2%). Content of Cd (0.23 +/- 0.13 mg/kg wet weight) and Hg (2.85 +/- 1.22 mg/kg wet weight) in the fish muscle exceeded maximum allowed levels in all samples. On average, the order of metal concentrations in the fish muscle was: Fe > Zn > Hg > Pb > Cu > Mn > Cd > Cr > Ni > Co. PMID- 15792308 TI - Results 28 months following transpupillary thermotherapy for classic and occult choroidal neovascularization in patients with age-related macular degeneration. AB - BACKGROUND AND OBJECTIVE: To report the outcome of patients 28 months following treatment with transpupillary thermotherapy (TTT) for classic and occult choroidal neovascularization (CNV) secondary to age-related macular degeneration. PATIENTS AND METHODS: A nonrandomized pilot trial of 36 eyes of 33 patients was performed. Eyes with angiographically defined CNV, 11 predominantly classical and 25 predominantly occult, were treated with large spot diode laser (810 nm) TTT for 1 minute, the end point being no or minimal visible change. Outcome was assessed with best-corrected LogMAR visual acuity, clinical examination, and fluorescein angiography. RESULTS: Patients were observed for a mean of 28.7 months (range, 18 to 40 months). The mean change in LogMAR visual acuity for predominantly classic membranes was -1.91 (standard deviation [SD] = 4.3) and 5 of 11 (45.5%) eyes had a loss of 3 or more LogMAR lines. Predominantly classic membranes were closed in 9 of 11 eyes and stabilized in 2 of 11 eyes. The mean change in LogMAR visual acuity for predominantly occult membranes was -1.48 (SD = 6.3) and 10 of 25 (40%) patients had a loss of 3 lines or more. Predominantly occult CNV was stabilized in 25 of 25 cases, and recurrence developed in 2 of 25 cases; one of the latter developed classic CNV. CONCLUSIONS: The medium-term results for patients treated with TTT for both occult and classic CNV show good stability, with little visual loss and few recurrences. These data confirm the original findings of this study. PMID- 15792309 TI - Thrombocytopenia and the risks of intraocular surgery. AB - BACKGROUND AND OBJECTIVE: In thrombocytopenia, a hemorrhagic diathesis is usually present due to a low platelet count and has been related to the development of cataracts and retinopathy. The concomitant administration of nonsteroidal anti inflammatory drugs can increase the hemorrhagic diathesis. The purpose of this study was to investigate the impact of thrombocytopenia during and after intraocular surgery. PATIENTS AND METHODS: A retrospective study of medical files of patients who had undergone cataract and glaucoma filtering surgery and were diagnosed as having thrombocytopenia between 1994--1995 and 1998--1999 was conducted. Eight patients with a total of 11 surgical procedures were included in this study. RESULTS: Hemorrhagic complications occurred in 2 of the 11 procedures, for an incidence of 18%. These 2 cases are described in detail. CONCLUSIONS: The current study confirms that thrombocytopenia is a significant risk factor for perioperative bleeding in ocular surgery. A routine questionnaire should be completed before ocular surgery and a complete blood cell count obtained in suspect cases. Consultation with a hematologist is warranted in cases of thrombocytopenia. PMID- 15792310 TI - Comparison of phacoemulsification and planned extracapsular cataract extraction in combined pars plana vitrectomy and posterior chamber intraocular lens implantation. AB - BACKGROUND AND OBJECTIVE: To compare two kinds of cataract removal methods combined with pars plana vitrectomy and posterior chamber intraocular lens (PC IOL) implantation. PATIENTS AND METHODS: Cataract removal was performed by either phacoemulsification or extracapsular cataract extraction (ECCE) and followed by pars plana vitrectomy and PC-IOL implantation. Between May 1996 and June 2001, 31 and 22 patients with cataract and vitreoretinal disease were treated by phacoemulsification and ECCE, respectively, combined with pars plana vitrectomy and PC-IOL implantation. Preoperative demographic data and postoperative vision, astigmatism change, and complications were analyzed. RESULTS: The mean age of patients in the phacoemulsification and ECCE groups was 62.5 and 63.4 years, respectively. Diabetes mellitus was the most common underlying disease resulting in vitreous opacity. Vision improved in 87.1% of the phacoemulsification group and 59.1% of the ECCE group. The change in astigmatism after surgery was 0.92+/ 1.08 D (P = .001) in the ECCE group and 0.25+/-0.74 D (P = .087) in the phacoemulsification group. There were fewer postoperative complications, including recurrent vitreous hemorrhage, increased intraocular pressure, and iris changes in the phacoemulsification group than in the ECCE group. CONCLUSION: Phacoemulsification and ECCE combined with pars plana vitrectomy and PC-IOL implantation are both effective surgical methods to achieve better and more rapid visual rehabilitation for patients with combined cataract and vitreoretinal disease. In this study, phacoemulsification combined with pars plana vitrectomy and PC-IOL implantation resulted in greater improvement in vision, less astigmatism change, and fewer postoperative complications. PMID- 15792312 TI - Automated irrigation/aspiration before phacoemulsification in eyes with white cataracts. AB - BACKGROUND AND OBJECTIVE: A white mature cataract still presents a significant challenge for the cataract surgeon. Because of the reduced visibility and increased intracapsular pressure in such cases, it is difficult to consistently achieve controlled continuous curvilinear capsulorhexis. Several simple procedures for treatment of eyes with white mature cataract are described. PATIENTS AND METHODS: Seven consecutive patients (7 eyes) with white cataracts were enrolled for cataract surgery from July 2002 to February 2003. A small, central anterior capsular opening was created after carefully filling the anterior chamber with viscoelastic agents. Automated irrigation/aspiration by lowering of a connected bottle of balanced salt solution was used to aspirate the liquefied milky lens contents and decrease intracapsular pressure. RESULTS: All patients obtained controlled capsulorhexis, safe cataract phacoemulsification, and in-the-bag intraocular lens implantation. CONCLUSION: Automated irrigation/aspiration before phacoemulsification is safe and effective for eyes with white cataracts. PMID- 15792311 TI - Unilateral and bilateral lateral rectus recession in exotropia. AB - BACKGROUND AND OBJECTIVE: To evaluate the surgical results obtained by unilateral and bilateral lateral rectus recession for the correction of exotropia. PATIENTS AND METHODS: The charts of all patients with exotropia who were operated on at the Goldschleger Eye Institute at Sheba Medical Center during an 11-year period were retrospectively reviewed. Study participants all underwent a complete orthoptic and ocular examination. Twenty-five patients with moderate-angle exotropia underwent unilateral lateral rectus recession (group 1) and 38 patients with large-angle exotropia underwent bilateral lateral rectus recession (group 2). The angle of exotropia was measured by the prism and cover test. Moderate exotropia was defined as 25 prism diopters (PD) and large-angle exotropia as greater than 25 PD. RESULTS: The mean age at the time of the surgery was 10.0+/ 5.2 years in group 1 and 8.5+/-8.0 years in group 2. The mean preoperative exotropia was 16.1+/-5.7 PD in group 1 and 29.6+/-14.4 PD in group 2. A mean postoperative exodeviation of 4.2+/-5.4 PD was found in group 1 and 5.8+/-13.6 PD in group 2. The success rate (deviation of < 10 PD) was 84% in group 1 and 74% in group 2. There was no incomitance in group 1. CONCLUSION: Unilateral lateral rectus recession is an effective surgical method for correcting moderate-angle exotropia with results similar to bilateral lateral rectus recession for larger exotropia angles. PMID- 15792313 TI - Regional anesthesia with monitored anesthesia care for surgical repair of selected open globe injuries. AB - BACKGROUND AND OBJECTIVE: To investigate clinical features and visual acuity outcomes associated with the use of regional anesthesia with monitored anesthesia care (RA/MAC) versus general anesthesia for open globe injuries in adult reparable eyes. PATIENTS AND METHODS: Retrospective, consecutive case series including all adults with open globe injuries repaired at Bascom Palmer Eye Institute between 2000 and 2003. RESULTS: The study included 238 eyes of 238 patients with a median age of 41 years. RA/MAC was employed in 141 (59%) eyes and general anesthesia in 97 (41%) eyes. Patients who had RA/MAC were significantly (P< .001) more likely to have anterior wound location (64% corneal/limbal vs 50%), shorter wound length (6.5 vs 12 mm), and formed anterior chamber (82% vs 56%), and were significantly less likely to have an afferent pupillary defect (12% vs 43%). The improvement from presenting visual acuity to each postoperative follow-up interval was similar for the two groups. CONCLUSIONS: RA/MAC is a reasonable alternative to general anesthesia for selected patients with open globe injuries. PMID- 15792314 TI - Ultrasound biomicroscopy features of iris and ciliary body melanomas before and after brachytherapy. AB - BACKGROUND AND OBJECTIVE: To determine ultrasound biomicroscopy (UBM) features of iris and ciliary body melanomas before and after brachytherapy. PATIENTS AND METHODS: Four uveal anterior melanoma cases undergoing brachytherapy were retrospectively studied. All cases were examined by UBM prior to treatment and repeatedly after treatment. RESULTS: Before brachytherapy, UBM examination showed a solid mass in the iris, ciliary body, or both in all four cases and allowed its characterization, sizing, and positioning. Two cases had a pigmented scleral lesion corresponding to the tumor location, but UBM did not detect any scleral infiltration at those sites. After brachytherapy, all lesions showed progressive decrease in size and progressive attenuation of their limits. Internal reflectivity was variable. Complications related to brachytherapy were demonstrated, including cataract, peripheral anterior synechiae at tumor location, and secondary scleral thinning. CONCLUSION: UBM played an important role as a complementary diagnostic method for anterior uveal melanomas, particularly ciliary body melanomas, but also allowed therapeutic planning (brachytherapy or surgery) and follow-up after treatment. PMID- 15792315 TI - Wide-field handheld high frequency ultrasonography using a variable high frequency ultrasound system. AB - Wide-field handheld high frequency ultrasonography using a variable high frequency ultrasound system is an imaging modality in the frequency range of 35 to 50 MHz. Using a water bath immersion technique, high-resolution digital movies obtained from a variety of anterior segment ocular conditions were particularly helpful in establishing the diagnosis in each case. Images taken with the wide field limbus-to-limbus mode provided a comprehensive picture of large and extensive lesions, whereas the focal mode was for small to medium lesions or specific areas of interest. PMID- 15792316 TI - Optic nerve measurements by 3D ultrasound-based coronal "C-scan" imaging. AB - Twenty-three normal eyes were examined with the Intensity Profiling technique of 3D I-Scan and 52 normal eyes with the Automated technique of OTI-Scan 1000 ultrasound systems (Ophthalmic Technologies Inc., Toronto, Ontario, Canada). With the eye looking straight, the probe was applied on the temporal sclera. Scanning generated 3D image files. Coronal optic nerve measurements were obtained 3 mm behind the globe. The mean optic nerve sheath diameter was 4.8 mm (standard deviation = 0.6; range, 3.9 to 5.9 mm) with the Intensity Profiling technique, and 5.4 mm (standard deviation = 0.4; range, 4.4 to 6.0 mm) with the Automated technique. 3D ultrasound imaging can be used to obtain optic nerve measurements in vivo. Both the Intensity Profiling and the Automated techniques yielded measurements similar to current magnetic resonance imaging and computed tomography scan reports. Coronal "C-scan" sectioning can be used as a screening tool to measure optic nerve diameter prior to magnetic resonance imaging or computed tomography. PMID- 15792317 TI - Optical coherence tomography appearance of "drusenoid" pigment epithelial detachment. AB - A case of bilateral "drusenoid" pigment epithelial detachment that was studied with fundus fluorescein angiography and optical coherence tomography is described. Fundus fluorescein angiography depicted staining of the drusen without any sign of pooling of the dye, whereas optical coherence tomography displayed detachment of the retinal pigment epithelium. These findings remained unchanged during a 10-month period. PMID- 15792318 TI - Late isolated brain metastasis following enucleation for choroidal melanoma. AB - Brain metastases from choroidal melanoma are rare and usually have a grave prognosis. A case of successfully treated late isolated brain metastasis from choroidal melanoma is described. A 35-year-old man presented with epileptic seizures of recent origin, 9 years following enucleation for choroidal melanoma. Imaging studies revealed a lesion of the right frontal lobe that was surgically removed. Results of pathologic examination were compatible with metastatic choroidal melanoma. The patient is asymptomatic 5 years postoperatively. Late isolated brain metastases from uveal melanoma may be treatable by local resection. Close, lifelong follow-up is required to diagnose and aggressively treat metastatic disease. PMID- 15792319 TI - Evolution of a retinoma case in 21 years. AB - A 35-year-old man was observed during a 21-year period with fundus color photography, fluorescein angiography, and B-scan ultrasonography for a presumed retinoma detected in his left eye during routine ophthalmoscopy in 1982. The patient was a carrier of the 13q14 retinoblastoma gene. His retinal lesion remained stable during the follow-up period without signs of malignant transformation. Retinomas are most commonly observed in patients with retinoblastoma and their relatives. In this case, none of the patient's parents, siblings, or children had evidence of a retinal tumor. Photographic documentation of the nonprogressive nature of this presumed retinoma is provided and demonstrates the absence of growth. PMID- 15792320 TI - HLA-B27-associated uveitis with a chorioretinitis manifestation. AB - A 29-year-old man who had been treated for acute anterior uveitis in a local medical office and observed for 1 month presented complaining of distorted vision in his left eye for 1 week. On ophthalmic examination, the anterior segment was relatively quiet with few cells. A posterior segment examination revealed cystoid macular edema and multiple splinter retinal hemorrhages. Results of all laboratory and imaging studies were negative, except for a positive HLA-B27 haplotype. Fluorescein angiography revealed massive leakage in the mid and late phase, consistent with chorioretinitis. Periocular corticosteroid injections and oral prednisolone were administered. The patient responded to the treatment well with subsequent resolution of chorioretinitis 2 months later. Although rare, chorioretinitis can occur in the setting of uveitis associated with HLA-B27 and seems to respond well to corticosteroid treatment. PMID- 15792321 TI - Indocyanine green angiography findings in sympathetic ophthalmia. AB - A 78-year-old woman with a recurrent retinal detachment in the right eye presented 2 years after her last surgery with clinical and fluorescein angiographic evidence of sympathetic ophthalmia in the left eye. Fluorescein angiography showed discrete multifocal areas of central hypofluorescence with a hyperfluorescent ring. After immunosuppressive therapy, fluorescein angiography displayed multiple hypofluorescent spots without the hyperfluorescent collar. Prior to systemic and periocular corticosteroid therapy, indocyanine green angiography (ICGA) revealed multifocal hypo-fluorescent spots that became more prominent as the study progressed. The early stages of the posttreatment ICGA appeared normal, but the hypofluorescent spots reappeared in the late stage of ICGA. ICGA is a useful diagnostic adjunct to fluorescein angiography and clinical examination in helping to secure the diagnosis and monitor the treatment progress of sympathetic ophthalmia. PMID- 15792322 TI - Management of glaucoma drainage implant tube kink and obstruction with Pars Plana Clip. AB - Increased intraocular pressure after glaucoma drainage implant surgery may be caused by obstruction of the tube. A case of obstruction of an Ahmed glaucoma valve tube after pars plana insertion due to kinking of the tube was treated with a Pars Plana Clip (New World Medical, Rancho Cucamonga, CA). PMID- 15792323 TI - Visualization of residual perfluorocarbon liquid using intravitreal triamcinolone acetonide. AB - The visualization of transparent perfluorocarbon liquid (PFCL) using triamcinolone acetonide is described. Intravitreal injection of triamcinolone acetonide enabled visualization of residual PFCL intraoperatively. In addition, it was shown that triamcinolone acetonide could visualize PFCL in an in vitro preparation of balanced salt solution. This in vitro experiment confirmed that triamcinolone acetonide also could be adsorbed by PFCL outside the vitreous. Triamcinolone acetonide was helpful to visualize transparent PFCL both in vivo and in vitro, and may be useful at the end of vitrectomy to completely remove residual PFCL from the eye. PMID- 15792324 TI - Intraocular segment of the central retinal vein. PMID- 15792325 TI - Impact of Iowa case management on provisions of social support for substance abuse clients. AB - Substance abuse can have a devastating effect on the social support networks of individuals. This article describes the impact of a comprehensive model of case management on substance abuse treatment clients'perceptions of social support. Social support was defined using a well-documented typology identifying six provisions for social support. The Iowa Case Management Project (ICMP) for Rural Drug Abuse was a randomized clinical trial using a longitudinal design. Clients were recruited from the residential treatment program of one rural Midwestern substance abuse treatment agency and were randomly assigned to one of three experimental conditions that received case management according to a strengths based model or to a control group. Client perceptions of social support were measured up to 12 months after treatment initiation. Results indicated that case management positively impacted overall perceptions of social support subsequent to treatment admission, as well as perceptions of support in two of the six provisions of support. Reassurances of worth, which is the recognition of one's competence, skills, and value by others, and attachment, which reflects close emotional bonds with others, were both significantly improved for clients who received case management. However, the effectiveness of case management in improving social support appears to be limited to clients who are married or have significant partners. PMID- 15792326 TI - Domestic violence: a review for case managers of regulations, women's center services, and referral questions. AB - The need for knowledge about domestic violence is woven into the case management's aim to strike a balance between quality and cost and its goal to coordinate care with community providers. The purpose of this article is to provide information for case managers on the current regulations that influence service delivery to victims of domestic battery, to provide an understanding of services provided by women's centers, and to understand referral questions that may support referrals to domestic violence services. PMID- 15792327 TI - Evaluation of perinatal care management programs: an integrated review. AB - Nursing care management has become a popular method to integrate health care systems with goals of decreasing costs and improving quality. As high-risk pregnancies and newborn intensive care unit (NBICU) costs generate some of the highest costs in health care, care management has been a strategy introduced in perinatal medicine to accomplish the same goals. Consistent with other areas of nursing, perinatal care management currently has no agreed upon model of practice or method of evaluating how and whether the goals have been achieved. The purpose of this project was to evaluate various perinatal care management programs found in the literature. Electronic and manual searches of current data were performed locating 31 relevant articles. From these articles, nine met the inclusion criteria and were reviewed. While some care management programs were able to decrease costs and improve outcomes, other programs did not. There are possibly two reasons for the varied results in the literature. The first is that there are differences in program designs, decreasing the likelihood of identifying specific interventions that can make a difference. Second, thus far researchers have used ineffective study designs in evaluating care management programs. More research needs to be completed before a conclusion can be drawn whether perinatal care management can decrease costs while improving quality. PMID- 15792328 TI - Long-term care insurance: care management practice issues. AB - There are numerous articles about the pros and cons of long-term care insurance. However, for many people it is a solution for funding long-term care when they are in need of these services. The provision of appropriate care management to individuals, providers, and insurers results in a "win-win" for everyone. The article discusses the issues surrounding long-term care financing and delivery from the perspective of the groups mentioned previously. PMID- 15792329 TI - Vision and hearing loss in older adults: "Double trouble". AB - Recent studies indicate that by age seventy, 21% of the people living in the United States have both vision and hearing loss. Dual sensory loss in the elderly has a significant effect on an individual's ability to socialize, communicate with others, and live independently. This article addresses the issues faced by older individuals who are hard of hearing and blind or visually impaired. Common causes and behavioral signs of hearing and vision loss are discussed. An emphasis is placed on the functional implications of the dual sensory impairment and possible accommodations and communication strategies are outlined PMID- 15792330 TI - Healing presence. AB - Medicine is the art and science of restoring and preserving health through the healing process. The science of medicine lies in evidenced based practices and the technological achievements of the last century, which enable practitioners to preserve the physical body. A growing number of research articles and writings have also appeared addressing the spiritual dimension of healing (the art of medicine), which often ignored creates a gap in the medical care of the patient. The spiritual aspects of medical care are explored based on the experience of the doctor-patient encounter. Three case studies of various clinical settings are presented to begin to look at the spiritual aspect of healing and better define its incorporation in the encounters with patients. The author reflects on the skills and concepts that begin to define the spiritual dimension of care: communication (listening, speaking), connection (space, safety, and sacredness), and communion through which healing can occur. As health care professionals we can be a "healing" (Hopeful, Empathetic, Attentive, Loving, Intentional, Nourishing, Grateful) presence taking care of the physical, mental and spiritual needs of our patients in our encounters with them, bringing them back to health and wholeness. PMID- 15792331 TI - Culture and long-term care: the bath as social service in Japan. AB - A central feature of Japan's approach to community-based care of the elderly, including long-term home health care, is the emphasis on providing bath facilities. For mobile elders, senior centers typically provide a public bathing facility in which people can enjoy a relaxing soak along with friends who also visit the centers. In terms of in-home long-term care, visiting bath services are provided to assist family care providers with the difflcult task of bathing a frail or disabled elder--a task made more problematic as a result of the Japanese style of bathing. I argue that the bath, as social service, is a culturally shaped solution to a specific problem of elder care that arises in the Japanese context as a result of the importance of the bath in everyday life for Japanese. While the services may be considered specific to Japan, some aspects of bathing services, particularly the mobile bath service, may also have applicability in the United States. PMID- 15792332 TI - Your ACHE partners at the local level. ACHE Regents and chapters are here to serve you. PMID- 15792333 TI - Interview with ACHE's incoming chairman. PMID- 15792334 TI - Improving ICU care: it takes a team. PMID- 15792335 TI - Take the lead or take your chances: engaging physicians in pay-for-performance. PMID- 15792337 TI - Organizational dynamics. The truth about creating a coaching culture. PMID- 15792336 TI - Organizational dynamics. Encouraging work/life balance. PMID- 15792338 TI - Recruitment and retention. Five questions to ask new hires. PMID- 15792339 TI - Making decisions about neonatal life support. Respecting the views of all concerned requires coordinated communication among all parties. PMID- 15792340 TI - Maximizing return on marketing resources. A data-driven process focuses marketing efforts on the right service lines. PMID- 15792341 TI - The right level of care at the right location. A Texas hospital addresses pediatric ED overutilization. PMID- 15792342 TI - Malpractice reform and the 109th Congress. The possibility for sweeping medical malpractice reform remains dim. PMID- 15792343 TI - Hitting the job search wall. Patience and persistence will help you reenergize your search. PMID- 15792344 TI - Physicians on the board. Understanding their delicate position can help you maximize physician trustee contributions. PMID- 15792345 TI - Adopting cross-industry best practices. A provider organization's case study demonstrates measurable results. PMID- 15792346 TI - Getting the most from your capital investments. A high ROI requires that you invest in staff education. PMID- 15792347 TI - Advancement to Diplomate or Fellow. Chapters can encourage members to take the next step in ACHE. PMID- 15792348 TI - Reduction of myocardial infarct size by doxycycline: a role for plasmin inhibition. AB - Myocardial ischemia-reperfusion (I/R) is associated with the activation of matrix metalloproteinases (MMPs) and serine proteases. We hypothesized that activation of MMPs and the serine protease plasmin contribute to early cardiac myocyte death following I/R and that broad-spectrum protease inhibition with doxycycline (DOX) preserves myocyte viability. Rats treated daily with or without DOX beginning 48 h prior to experimentation were subjected to 30 min of coronary occlusion and 2 days of reperfusion. DOX pre-treatment reduced infarct size by 37%. DOX attenuated increases in MMP-9 and plasmin levels as determined by gelatin zymography and immunoblot, respectively. Neutrophil extravasation was unaltered by DOX as assessed by myeloperoxidase (MPO) activity. To examine the contribution of MMP-9 and plasmin to myocyte injury, cultures of neonatal rat ventricular myocytes (NRVMs) were treated for 48 h with 83 kDa MMP-9 or plasminogen in the presence or absence of DOX. MMP-9 treatment did not affect myocyte viability. Plasminogen treatment led to increased plasmin activity, resulting in loss of beta1-integrin, NRVM detachment and apoptosis. DOX co-treatment inhibited plasmin activity and preserved NRVM attachment, whereas co-treatment with the broad spectrum MMP inhibitor GM6001 had no effect. These results indicate that plasmin causes disruption of myocyte attachment and viability independently of MMP activation in vitro and that inhibition of plasmin by DOX may reduce I/R-induced myocyte death in vivo through the inhibition of plasmin. PMID- 15792349 TI - Induction of caspase-independent apoptosis in H9c2 cardiomyocytes by adriamycin treatment. AB - The cardiotoxicity of adriamycin limits its clinical use as a powerful drug for solid tumors and malignant hematological disease. Although the precise mechanism by which it causes cardiac damage is not yet known, it has been suggested that apoptosis is the principal process in adriamycin-induced cardiomyopathy, which involves DNA fragmentation, cytochrome C release, and caspase activation. However, there has been no direct evidence for the critical involvement of caspase-3 in adriamycin-induced apoptosis. To determine the requirements for the activation of caspase-3 in adriamycin-treated cardiac cells, the effect of a caspase inhibitor on the survival of and apoptotic changes in H9c2 cells was examined. Exposure of H9c2 cells to adriamycin resulted in a time- and dose dependent cell death, and the cleavage of pro-caspase-3 and of the nuclear protein poly (ADP'ribose) polymerase (PARP). However, neither the reduction of cell viability nor the characteristic morphological changes induced by adriamycin were prevented by pretreatment with the general caspase inhibitor z-VAD.FMK. In contrast, caspase inhibition effectively blocked the apoptosis induced by H202 in H9c2 cells, as determined by an MTT assay or microscopy. We also observed that p53 expression was increased by adriamycin, and this increase was not affected by the inhibition of caspase activity, suggesting a role for p53 in adriamycin induced caspase-independent apoptosis in cardiac toxicity. PMID- 15792350 TI - p53 and cell-cycle-regulated protein expression in small intestinal cells after fast-neutron irradiation in mice. AB - The involvement of the p53 gene in apoptosis of many cell types towards y radiation is well established. However, little information is available on the relationship between p53 status and cells' ability to undergo apoptosis following exposure to fast neutrons. The aim of this study was to characterize the apoptotic pathway traveled by neutrons in mouse intestinal crypt cells. Each mouse received whole body doses of 0.25-8 Gy fast neutrons and were sacrificed 0, 4, 6, 12, 24, 48, and 72 h, respectively, after irradiation. Apoptosis of crypt cells and expression of p53, cyclin A, cyclin B, cyclin D, and cyclin E were measured. The apoptosis in crypt cells was maximal at 4 and 6 h after irradiation, showing a gradual decline at 24 h. The highest frequency of apoptosis was seen at a 1 Gy dose and then declined gradually beyond a 2 Gy dose with high levels of damage. In immunoblot analysis, apoptosis was confirmed to be dependent on p53 function after fast-neutron irradiation. In addition, cyclin B1, cyclin D, and cyclin E were overexpressed in intestinal cells after fast-neutron irradiation and their immunoreactivities were increased strongly in round and oval cells of laminar propria in villi core and crypts. The results of the current study suggest that apoptosis in crypt cells shows a time- and dose dependent increase after fast-neutron irradiation. In addition, fast-neutron induced apoptosis in mouse intestinal crypt cells appears to be related to the increase in functional p53 proteins to a level sufficient to initiate apoptosis and up-regulation of cell-cycle-regulated proteins, which may lead to resistance to DNA damage through cell cycle arrest, is involved deeply in protection of gastrointestinal cells after low doses of fast-neutron irradiation. PMID- 15792351 TI - The protective role of DL-alpha-lipoic acid in the oxidative vulnerability triggered by Abeta-amyloid vaccination in mice. AB - Recent reports indicate that beta-amyloid peptide (Abeta) vaccine based therapy for Alzheimer's disease (AD) may be on the horizon. There are however, concerns about the safety of this approach. Immunization with Abeta has several disadvantages, because it crosses the blood brain barrier and cause inflammation and neurotoxicity. The present work is aimed to study the protective effective of alpha-lipoic acid (LA) in the oxidative vulnerability of beta-amyloid in plasma, liver, spleen and brain, when Abeta fibrils are given intraperitoneally in inflammation induced mice. Result shows that reactive oxygen species (ROS) in the astrocytes of inflammation induced mice along with Abeta (IA) has shown 2.5-fold increase when compared with LA treated mice. The increased level of lipid peroxidase (LPO) (p < 0.05) and decreased antioxidant status (p < 0.05) were observed in the plasma, liver, spleen and brain of LA induced mice when compared with LA treated mice. Data shows that there were no significant changes observed between the control and LA treated mice. Our biochemical and histological results highlight that significant oxidative vulnerability was observed in IA treated mice, which was prevented by LA therapy. Our findings suggest that the antioxidant effect of LA when induced with Abeta may serve as a potent therapeutic tool for inflammatory AD models. PMID- 15792352 TI - Hierarchical change in antioxidant enzyme gene expression and activity in acute cardiac rejection: role of inducible nitric oxide synthase. AB - Reactive oxygen and nitrogen may mediate inflammation injury, but the status of the antioxidant defense system that might influence this process is unknown. In the present study, we examined the expression profile of the antioxidant enzymes, manganese superoxide dismutase (MnSOD), catalase and glutathione peroxidase (GPX) in acutely rejecting cardiac allografts and the potential role of inducible nitric oxide synthase (iNOS) in modulating antioxidant gene expression and activity. Donor hearts from Lewis (isograft) or Wistar-Furth (allograft) rats were transplanted into Lewis recipient rats. A subset of the allografts received L-N6-(1-imino-ethyl) lysine (L-NIL), a specific iNOS inhibitor, beginning the day of surgery until the day of harvesting. Catalase and glutathione peroxidase (GPX) protein levels were significantly decreased by postoperative day 4 (POD4) and postoperative day 5 (POD5), respectively, in allografts compared to isografts. While CuZn superoxide dismutase (CuZn SOD) levels were unchanged, there was a 50% decrease in MnSOD protein in allografts at postoperative day 6 (POD6). The sequential loss in antioxidant protein levels was not due to transcriptional regulation since there was no change in RNA levels for any of the genes tested. L NIL did not alter catalase protein; however, the loss of MnSOD protein at POD6 was prevented by L-NIL. Consistent with a decrease in antioxidant protein levels, there was a sequential loss in enzyme activity for MnSOD, catalase and GPX. L-NIL however, restored MnSOD and GPX activities but not catalase activity. Treatment with CsA restored both protein and enzyme activities of GPX and MnSOD but not catalase. These results indicate that the loss in MnSOD and GPX protein and activity in allografts occurs via an iNOS-dependent mechanism whereas the decrease in catalase appears to be iNOS-independent. This suggests a differential role for iNOS in regulating post-translational modification of individual antioxidant enzymes in acute cardiac transplantation. PMID- 15792354 TI - Protein kinase C isozymes in hypertension and hypertrophy: insight from SHHF rat hearts. AB - Chronic hypertension results in cardiac hypertrophy and may lead to congestive heart failure. The protein kinase C (PKC) family has been identified as a signaling component promoting cardiac hypertrophy. We hypothesized that PKC activation may play a role mediating hypertrophy in the spontaneously hypertensive heart failure (SHHF) rat heart. Six-month-old SHHF and normotensive control Wistar Furth (WF) rats were used. Hypertension and cardiac hypertrophy were confirmed in SHHF rats. PKC expression and activation were analyzed by Western blots using isozyme-specific antibodies. Compared to WF, untreated SHHF rats had increased phospho-active alpha (10-fold), delta (4-fold), and epsilon (3 fold) isozyme expression. Furthermore, we analyzed the effect of an angiotensin II type 1 receptor blocker (ARB) and hydralazine (Hy) on PKC regulation in SHHF rat left ventricle (LV). Both the ARB and Hy normalized LV blood pressure, but only the ARB reduced heart mass. Neither treatment affected PKC expression or activity. Our data show differential activation of PKC in the hypertensive, hypertrophic SHHF rat heart. Regression of hypertrophy elicited by an ARB in this model occurred independently of changes in the expression and activity of the PKC isoforms examined. PMID- 15792353 TI - Compartmentation of energy metabolism in atrial myocardium of patients undergoing cardiac surgery. AB - The parameters of oxidative phosphorylation and its interaction with creatine kinase (CK)- and adenylate kinase (AK)-phosphotransfer networks in situ were studied in skinned atrial fibers from 59 patients undergoing coronary artery bypass surgery, valve replacement/correction and atrial septal defect correction. In atria, the mitochondrial CK and AK are effectively coupled to oxidative phosphorylation, the MM-CK is coupled to ATPases and there exists a direct transfer of adenine nucleotides between mitochondria and ATPases. Elimination of cytoplasmic ADP with exogenous pyruvate kinase was not associated with a blockade of the stimulatory effects of creatine and AMP on respiration, neither could it abolish the coupling of MM-CK to ATPases and direct transfer of adenine nucleotides. Thus, atrial energy metabolism is compartmentalized so that mitochondria form functional complexes with adjacent ATPases. These complexes isolate a part of cellular adenine nucleotides from their cytoplasmic pool for participating in energy transfer via CK- and AK-networks, and/or direct exchange. Compared to atria in sinus rhythm, the fibrillating atria were larger and exhibited increased succinate-dependent respiration relative to glutamate dependent respiration and augmented proton leak. Thus, alterations in mitochondrial oxidative phosphorylation may contribute to pathogenesis of atrial fibrillation. PMID- 15792355 TI - Energy metabolism in the granulation tissue of diabetic rats during cutaneous wound healing. AB - The skin cells chiefly depend on carbohydrate metabolism for their energy requirement during cutaneous wound healing. Since the glucose metabolism is greatly hampered in diabetes and this might affect wound repair process. This prompted us to investigate the intermediate steps of energy metabolism by measuring enzyme activities in the wound tissues of normal and streptozotocin induced diabetic rats following excision-type of cutaneous injury. The activities of key regulatory enzymes namely hexokinase (HK), phosphofructokinase (PFK), lactate dehydrogenase (LDH), citrate synthase (CS) and glucose-6 phosphate dehydrogenase (G6PD) have been monitored in the granulation tissues of normal and diabetic rats at different time points (2, 7, 14 and 21 days) of postwounding. Interestingly, a significant alteration in all these enzyme activities was observed in diabetic rats. The activity of PFK was increased but HK, LDH and CS showed a decreased activity in the wound tissue of diabetics as compared to normal rats. However G6PD exhibited an elevated activity only at early stage of healing in diabetic rats. Thus, the results suggest that significant alterations in the activities of energy metabolizing enzymes in the wound tissue of diabetic rats may affect the energy availability for cellular activity needed for repair process and this may perhaps be one of the factor responsible for impaired healing in these subjects. PMID- 15792356 TI - Role of MMP-2 in inhibiting Na+ dependent Ca2+ uptake by H2O2 in microsomes isolated from pulmonary smooth muscle. AB - Treatment of microsomes (preferentially enriched with endoplasmic reticulum) isolated from bovine pulmonary artery smooth muscle tissue with H2O2 (1 mM) markedly stimulated matrix metalloproteinase activity and also inhibited Na+ dependent Ca2+ uptake. Electron micrograph revealed that H2O2 (1 mM) does not cause any damage to the microsomes. MMP-2 and TIMP-2 were determined to be the ambient protease and corresponding antiprotease of the microsomes. Pretreatment with vitamin E (1 mM) and TIMP-2 (50 microg/ml) reversed the effect produced by H2O2 (1 mM) on Na+ dependent Ca2+ uptake in the microsomes. However, H2O2 (1 mM) caused changes in MMP-2 activity and Na+ dependent Ca2+ uptake were not reversed upon pretreatment of the microsomes with a low concentration of 5 microg/ml of TIMP-2 which otherwise reversed MMP-2 (1 microg/ml) mediated increase in 14C gelatin degradation and inhibition of Na+ dependent Ca2+ uptake. Combined treatment of the microsomes with a low dose of MMP-2 (0.5 microg/ml) and H2O2 (0.5 mM) inhibited Na+ dependent Ca2+ uptake in the microsomes compared to the respective low dose of either of them. Direct treatment of TIMP-2 (5 microg/ml) with H2O2 (1 mM) abolished the inhibitory effect of the inhibitor on 14C gelatinolytic activity elicited by 1 microg/ml of MMP-2. Thus, one of the mechanisms by which H2O2 activates MMP-2 could be due to inactivation of TIMP-2 by the oxidant. The resulting activation of MMP-2 subsequently inhibits Na+ dependent Ca2+ uptake in the microsomes. PMID- 15792357 TI - Hypoxia and low glucose differentially augments TRAIL-induced apoptotic death. AB - Tumor microenvironment, which is characterized by hypoxia, low-glucose concentrations, high-lactate concentrations, low-extracellular pH, can alter the therapeutic response in tumors. In this study, we investigated whether hypoxia affects TRAIL-induced apoptotic death. When human prostate adenocarcinoma DU-145 cells were treated with 50 ng/mL TRAIL or hypoxia for 4 h, the survival was 45.7 and 32.5%, respectively. The combination of TRAIL and hypoxia synergistically increased cell death. Similar results were observed in human prostate adenocarcinoma LNCaP cells. Western blot analysis showed that the hypoxia augmented TRAIL-induced PARP cleavage as well as the activation of caspase-8 and caspase-3, but not caspase-9. Unlike hypoxia, low glucose promoted caspase-9 activation during TRAIL treatment. These results suggest that hypoxia or low glucose-augmented TRAIL cytotoxicity is mediated through the mitochondria independent pathway or -dependent pathway, respectively. PMID- 15792358 TI - Protein expression pattern of CDK11(p58) during testicular development in the mouse. AB - Protein kinases are important signalling molecules critical for normal cell growth and development. CDK11(p58) is a p34(cdc2) related protein kinase, and plays an important role in normal cell cycle progression. In this study, we mainly characterized the protein expression of CDK11(p58) during postnatal development in mouse testes and examined the cellular localization of CDK11(p58) and cyclinD3, which was associated with CDK11(p58) in mammalian cells. Western blot analysis revealed that CDK11(p58) was present in the early stages of development. It gradually increased and reached a peak in adult testes. The protein expression of CDK11(p58) was further analysed by immunohistochemistry due to its developmentally regulated expression. The variable immunostaining patterns of CDK11(p58) were visualized during different developmental periods and, in adult mouse, different stages of seminiferous tubules. CDK11(p58) expression was detected in proliferating germ cells in the early stages of developing testes. In adult testes, the protein was expressed in pachytene primary spermatocytes from stage VII to XI of spermatogenesis and in postmeiotic spermatids in all stages at different levels. The colocalization of CDK11(p58) and cyclinD3 in the adult testis was revealed by immunofluorescence analysis. PMID- 15792359 TI - The catalytically active secretory phospholipase A2 type IIA is involved in restenosis development after PTCA in human coronary arteries and generation of atherogenic LDL. AB - Secretory phospholipase A2 type IIA (sPLA2) may actively contribute to atherogenesis, acting either within the arterial wall or in plasma. Proinflammatory eicosanoids and lysophospholipids, generated through hydrolysis of cell membrane phospho-lipids by sPLA2, initiate and prolong the inflammatory process. In the present study we examined the possible involvement of sPLA2 in development of restenosis in patients undergoing percutaneous transluminal coronary angioplasty (PTCA). We also investigated whether serum sPLA2 could catalyze accumulation of lysophosphatidylcholine (LPC) in LDL. Concentrations and catalytic activities of sPLA2 were measured in blood serum of 49 consenting patients immediately before, 1-7 and 180 days after PTCA. All patients had repeat angiograms at 180-day follow-up. Restenosis was registered in 19 patients. Accumulation of LPC in LDL was evaluated by thin-layer chromatography after incubation of blood serum with LDL. Serum sPLA2 concentrations increased in all study patients by day 1 post-PTCA, but the increase was significantly greater and more protracted in patients who developed restenosis. Catalytic activities increased significantly 6 days post-PTCA in patients who developed restenosis, whereas for patients without restenosis there was no change in serum sPLA2 activity throughout the study period in spite of the sPLA2 presence in blood. Incubation of blood serum (6 days post-PTCA) with LDL resulted in accumulation of LPC only for those patients who subsequently developed restenosis. Manoalide, a specific inhibitor of sPLA2, completely blocked the LPC accumulation. The data indicate that elevated serum sPLA2 activity after PTCA is associated with restenosis development and may be involved in atherogenic modification of LDL in blood serum. PMID- 15792360 TI - Assessing the cellular transmembrane electrical potential difference on the hepatic uptake of palmitate. AB - Understanding the driving forces for the hepatic uptake of endogenous and exogenous substrates in isolated cells and organs is fundamental to describing the underlying hepatic physiology/pharmacology. In this study we investigated whether uptake of plasma protein-bound [3H]-palmitate across the hepatocyte wall is governed by the transmembrane electrical potential difference (PD). Uptake was studied in isolated hepatocytes and isolated perfused rat livers (IPL). Protein binding and vasoactive properties of the different perfusates were determined using in vitro heptane/buffer partitioning studies and the multiple indicator dilution (MID) technique in the IPL, respectively. Altering hepatocyte PD by perfusate ion substitution resulted in either a substantial depolarization (-14 +/- 1 mV, n = 12, mean +/- S.E., substituting choline for Na+) or hyperpolarization (-46 +/- 3 mV, n = 12, mean +/- S.E., substituting nitrate for Cl-). Perfusate ion substitution also affected the equilibrium binding constant for the palmitate-albumin complex. IPL studies suggested that, other than with gluconate buffer, hepatic [3H]-palmitate extraction was not affected by the buffer used, implying PD was not a determinant of extraction. [3H]-Palmitate extraction was much lower (p < 0.05) when gluconate was substituted for Cl- ion. This work contrasts with that for the extraction of [3H]-alanine where hepatic extraction fraction was significantly reduced during depolarization. Changing the albumin concentration did not affect hepatocyte PD, and [3H]-palmitate clearance into isolated hepatocytes was not affected by the buffers used. MID studies with vascular and extravascular references revealed that, with the gluconate substituted buffer, the extravascular volume possibly increased the diffusional path length thus explaining reduced [3H]-palmitate extraction fraction in the IPL. PMID- 15792361 TI - Antimicrobial effects of antioxidants with and without clarithromycin on Helicobacter pylori. AB - Increasing resistance to currently used antimicrobials has resulted in the evaluation of other agents that have antimicrobial activity against Helicobacter pylori. H. pylori American Type Culture Collection (ATCC) strain 49503 (a toxin producing strain known to be associated with gastric cancer) was grown, a cell suspension prepared in 2 mL PBS and diluted 10-fold. One hundred microL of this cell suspension was added to vitamin C 0.5%, vitamin E 0.5%, garcinol 100 microg/mL, Protykin (containing 50% trans-resveratrol) 100 microg/mL and garcinol + Protykin 100 microg/mL in Lennox broth, and incubated for 16 h under microaerophilic conditions. Three replicates of 10 microL from each 10(-7) dilution tube were plated, colonies were counted after 16 h, and growth of H. pylori was confirmed by the CLO test. These colony counts were compared to control cultures without the addition of any antioxidants. The experiments were then repeated with the addition of 15 microg/mL of clarithromycin to experimental and control samples. Enhanced killing of H. pylori by 37.6% was noted when vitamin C was added, which increased to 66% when clarithromycin was added, compared to controls (p < 0.05). With garcinol and Protykin alone there was 91.4 and 87% killing of H. pylori, respectively, while a combination of garcinol + Protykin resulted in 90.8% killing compared to controls (p < 0.05). When clarithromycin was added, there was 76.3% increased killing with garcinol alone, 55.3% with Protykin alone, and 73.7% with garcinol + Protykin compared to controls (containing clarithromycin) (p < 0.05). Vitamin E had no effect on H. pylori growth compared to controls. We conclude from this study that some antioxidants such as vitamin C, garcinol and Protykin, but not vitamin E, may have potential as antimicrobial agents against H. pylori. PMID- 15792362 TI - Effects of dietary selenium on post-ischemic expression of antioxidant mRNA. AB - Cardiac ischemia reperfusion leads to oxidative stress and poor physiological recovery. Selenium deficiency down-regulates thioredoxin reductase (Txnrd) and glutathione peroxidase (Gpx) activity, impairing recovery from ischemia reperfusion. Furthermore, selenium supplementation has been shown to be cardioprotective and lessens oxidative stress in reperfused rat hearts. In this study we have investigated the role of selenium in the mRNA expression of these, and related antioxidant proteins, post ischemia-reperfusion. Male rats were fed varying doses of selenium for five weeks. Hearts were isolated and perfused using the Langendorff method with 22.5 min of global ischemia and 45 min reperfusion. RNA was extracted for quantitative real-time PCR analysis of glutathione peroxidase (Gpx)-1 and 4, glutathione reductase (Gsr), thioredoxin peroxidase-2 (Prdx2), thioredoxin (Txn) and thioredoxin reductase (Txnrd)-1 and 2 gene expression. Selenium deficiency produced significant reductions in Gpx-1, Gpx-4, Prdx2, Txnrd-1 and Txnrd-2 expression. Conversely, selenium supplementation of 1000 microg/kg significantly up-regulated Gpx-1, Gpx-4, Txn, Txnrd-1 and Txnrd-2 transcription. Our results show selenium modulates the cardiac mRNA expression of thioredoxin and glutathione related enzymes post ischemia-reperfusion, and impacts on tolerance to ischemia-reperfusion. PMID- 15792363 TI - Effect of chronic administration of lignan complex isolated from flaxseed on the hemopoietic system. AB - Lignan complex has been isolated from flaxseed. It has been shown to reduce serum lipids and the extent of hypercholesterolemic atherosclerosis. However, it is not known whether the chronic use of lignan complex has any adverse effects on the hemopoietic system. The effects of lignan complex (40 mg/kg body wt orally daily for 2 months) on the red blood cells (RBC) count, mean corpuscular volume (MCV), red cell distribution width (RDW), hematocrit (Hct), hemoglobin (Hb), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and counts of white blood cell (WBC), granulocytes, lymphocytes, monocytes and platelet, and platelet volume were investigated in normo- and hypercholesterolemic rabbits. The results show that lignan complex had no adverse effects of counts of RBC, WBC, granulocytes, lymphocytes, monocytes and platelet in both the normo- and hyper-cholesterolemic rabbits. The values for MCV, RDW, Hct, Hb, MCH, MCHC, and platelet volume were similar in lignan complex-treated or untreated normo- and hypercholesterolemic rabbits. It is concluded that chronic use of lignan complex had no adverse effects on the hemopoietic system. PMID- 15792364 TI - Effects of chronic quercetin treatment in experimental renovascular hypertension. AB - The aims of the present study were to analyse the effects of an oral daily dose (10 mg/kg) of the dietary flavonoid quercetin for five weeks in two-kidney, one clip (2K1C) Goldblatt (GB) hypertensive rats. The evolution of systolic blood pressure was followed by weekly measurements, and morphological variables, proteinuria, plasma nitrates plus nitrites (NOx) and thiobarbituric acid reactive substances (TBARS), liver oxidative stress markers and endothelial function were determined at the end of the experimental period. Quercetin treatment reduced systolic blood pressure of GB rats, producing no effect in control animals. It also reduced cardiac hypertrophy and proteinuria developed in GB hypertensive rats. Decreased endothelium-dependent relaxation to acetylcholine of aortic rings from GB rats was improved by chronic quercetin treatment, as well as increased endothelium-dependent vasoconstrictor response to acetylcholine and overproduction of TXB2 by aortic vessels of GB rats, being without effect in normotensive animals. Increased plasma NOx and TBARS, and decreased liver total glutathione (GSH) levels and glutathione peroxidase (GPX) activity were observed in GB hypertensive rats compared to the control animals. Normalisation of plasma NOx and TBARS concentrations and improvement of the antioxidant defences system in liver accompanied the antihypertensive effect of quercetin. We conclude that chronic oral treatment with quercetin shows both antihypertensive and antioxidant effects in this model of renovascular hypertension. PMID- 15792365 TI - Paradoxical effects of hypoxia-mimicking divalent cobalt ions in human endothelial cells in vitro. AB - Divalent cobalt ions (Co2+) induce the expression of hypoxia responsive genes and are often used in cell biology to mimic hypoxia. In this in vitro study we compared the effects of hypoxia and Co2+ on human endothelial cells and examined processes that are stimulated in hypoxia in vivo (proliferation and angiogenesis). We analyzed the expression of the hypoxia-inducible factor-1alpha (HIF-1alpha) under different hypoxic conditions (3% and nearly 0% O2) and Co2+ concentrations (0.01-0.7 mM). As in hypoxia, the amount of HIF-1alpha protein was enhanced by exposure to Co2+ (did not correlate with mRNA amount). however, contrary to the results of hypoxia, in vitro-angiogenesis was inhibited after exposure to even low Co2+-concentrations (> or =0.01 mM). This led to the conclusion that although hypoxia signaling after Co2+ -exposure took place, further yet unknown Co2+ -induced event(s) must have occurred. PMID- 15792366 TI - Role of Salmonella surface components in immunomodulation of inflammatory mediators. AB - Salmonella enterica serovar Typhimurium and its surface components were assessed for their inflammatory potential by footpad oedema test using plethysmometer. Inflammation was found to be the highest when outer membrane proteins (OMPs) were used as inflammagen followed by lipid associated protein-lipopolysaccharide complex (LAP-LPS) and lipopolysaccharides (LPS). Inflammation produced by OMPs was found to be comparable to that by carrageenan (a known positive inflammagen). However, injection of L-histidine (an antioxidant) prior to administration of carrageenan or Salmonella enterica serovar Typhimurium inhibited the inflammation, which indicated the involvement of oxidants during inflammatory response. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and nitric oxide (NO) production by peritoneal macrophages from infected mice exhibited a significant increase as compared to those of the immunized mice. In contrast, glutathione production was found to be the maximum in the macrophages taken from OMPs-immunized mice followed by LAP-LPS and LPS alone. The biochemical studies correlated well with histopathological studies of intestinal tissue of animals from various groups. Based upon these parameters, inflammation seems to be modulated by OMPs and LAP-LPS, which may be because of the protein moieties present in the components. Hence, immunization with protein moieties having L histidine or L-histidine-like structures may suggest an alternative to the potential therapeutic values of anti-inflammatory drugs. Thus the results of this study form the basis for evaluating these antigens (either alone or in combination with polysaccharides) for preventive intervention rather than therapeutic. PMID- 15792368 TI - 5alpha-reductase in human embryonic kidney cell line HEK293: evidence for type II enzyme expression and activity. AB - Human 5alpha-reductase catalyses the last step in androgen biosynthesis, namely the reduction of testosterone (T) to the more potent androgen dihydrotestosterone (DHT). The enzyme is therefore considered to be an important drug target for androgen related diseases such as benign prostatic hyperplasia and prostate cancer. The present study displays evidence that the human embryonic kidney cell line HEK293 which is frequently used in recombinant target protein expression contains an endogenous 5alpha-reductase type II activity. After an incubation of 24 h 1 x 10(6) HEK293 cells converted 23% of the substrate 4-androstene-3,17 dione (7.5 nM) to the product 5alpha-androstane-3,17-dione. Reverse transcription polymerase chain reaction was carried out to identify the mRNA of the isoform responsible for the 5alpha-reductase activity. Only with type II specific primers a fragment with the predicted size was amplified, while with type I specific primers no band could be observed. An antiserum against human 5alpha-reductase type II was raised by immunizing a rabbit with a hemocyanin-conjugated peptide corresponding to amino acid 29 to 44 of the type II enzyme. Western blot analysis of different fractions of a HEK293 homogenate performed with this antiserum detected a band at 45 kDa in the nuclear and microsomal fraction corresponding to 5alpha-reductase type II protein. PMID- 15792370 TI - Alternative splicing generates a CaM kinase IIbeta isoform in myocardium that targets the sarcoplasmic reticulum through a putative alphaKAP and regulates GAPDH. AB - We report the isolation of a full length cDNA from cardiac muscle that encodes a approximately 73 kDa calcium/calmodulin (CaM) dependent kinase IIbeta isoform (CaMKIIbeta(C)) that was generated by alternative splicing of the CaMKIIbeta gene. Antipeptide antibodies raised to specific regions of the kinase identified a 73 kDa kinase polypeptide in cardiac SR. Anti-alpha kinase anchoring protein (alphaKAP) antibodies identified a 25 kDa polypeptide in cardiac SR and RT-PCR followed by sequence analysis confirmed the presence of a full length alphaKAP encoding transcript in myocardium. Protein interaction assays revealed that the 73 kDa CaMKIIbeta(C) binds GAPDH to modulate the production of NADH in a Ca2+/CaM dependent reaction. The presence of a CaMKIIbeta isoform that can target the SR presumably via its membrane anchor alphaKAP defines a previously unrecognized Ca2+/CaM regulatory system in myocardium. PMID- 15792371 TI - Effect of streptozotocin-induced diabetes on daily expression of per2 and dbp in the heart and liver and melatonin rhythm in the pineal gland of Wistar rat. AB - The circadian system is a flexible framework allowing a proper adjustment of physiological functions to the regularly changing environment. Pathways that are used to synchronize components of the circadian system have been shown to be susceptible to pathophysiological conditions. In our study, we investigated effects of streptozotocin (STZ)-induced diabetes mellitus on function of the circadian system at the level of melatonin synthesis and expression of per2 and dbp in the heart and liver in 8-week-old Wistar rats. Rhythmic pattern of clock gene per2 and transcription factor dbp in controls and STZ-treated animals was determined. Streptozotocin administration had a more substantial effect on per2 expression in the liver than in the heart. Pronounced phase advance in the rhythm of dbp expression in both the liver and the heart was observed. The melatonin rhythm reflecting the phase of the master clock was not affected by STZ application. Changes in per2 and dbp expression in the heart and liver imply alterations in input pathway or peripheral oscillators with possible consequences on function of analysed organs. PMID- 15792372 TI - [Bcl-2 in ovarian carcinoma: a clinicopathologic, immunohistochemical and molecular study]. AB - We investigated the immunohistochemical espression of bcl-2 and the genetic assessment of the bcl-2 gene in relation to responsiveness to first line chemotherapy and to the clinical outcome in advanced ovarian carcinoma patients. We have compared 17 patients, with FIGO stage III C, ovarian serous carcinomas, G3, living with no evident disease five years after primary surgical treatment; to 19 patients who had died of progression of disease no later than two years after primary surgical treatment. The correlation of bcl-2 expression with the survival and the clinical responsiveness to chemotherapy, were analysed with the logistic regression. We observed a bcl-2 expression in tumor cells in 25% of the cases. Molecular genetic analysis of the bcl-2 gene was performed for all the bcl 2 immunohistochimical positive cases. No traslocation t(14;18)(q32;q21) of the gene bcl-2 were found. The bcl-2 over-expression was found to be a significant independent predictor of responsiveness to chemotherapy (p = 0.04), but it was not correlated with the overall survival of the ovarian cancer patients. The prognostic value of bcl-2 espression may help in the management of ovarian cancer patients permitting the selection of more aggressive first line chemotherapy. In addition, the knowledge of the molecular mechanism, which is responsible of the over-expression of bcl-2, may help in the understanding of mechanisms responsible for chemoresistance. Further studies in this area will help clarify this therapeutic possibility. PMID- 15792369 TI - Interaction of exercise and adenosine receptor agonist and antagonist on rat heart antioxidant defense system. AB - This study investigated the interactive effects of acute exercise and adenosine receptor agonist and antagonist on antioxidant enzyme activities, glutathione and lipid peroxidation in the heart of the rat. Male Fisher-344 rats were divided into six groups and treated as follows: (1) saline control; (2) acute exercise (100% VO2max); (3) R-Phenyl isopropyl adenosine (R-PIA) (3.46 micromol/kg, i.p.); (4) theophylline (1.70 micromol/kg, i.p.) plus acute exercise; (5) theophylline plus R-PIA; and (6) theophylline. Animals were sacrificed 1 h after treatments; hearts were isolated and analyzed. The results show that acute exercise as well as adenosine receptor agonist R-PIA significantly enhanced cardiac superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione reductase (GR) activity by 36-135% and 16-51%, respectively. Adenosine receptor agonist R-PIA significantly decreased cardiac GSSG concentration and enhanced GSH/GSSG ratio by 22 and 30%, respectively. Whereas theophylline treatment blocked the activation of antioxidant enzyme activities enhanced by acute exercise and R-PIA. Theophylline treatment significantly increased lipid peroxidation by 43% in the heart of exercised rats. The study concluded that the adenosine receptors are involved in the upregulation of cardiac antioxidant defense system and attenuation of lipid peroxidation due to acute exercise in rats. PMID- 15792373 TI - [Tumoral calcinosis in a Sardinian patient mimicking hydatid disease: a case report]. AB - The aim of this study is to report on a case of tumoral calcinosis (TC) mimicking hydatid cyst, which was diagnosed in a 51-year-old Caucasian woman of Sardinian origin. This lady presented with two symmetrical enlarging masses of soft tissue in the hips. The CT findings were suggestive of hydatid cysts. Grossly the two lesions appeared as unencapsulated firm, rubbery masses extending into the surrounding muscles. On cut surface the two specimens were multiloculated and contained either calcareous material or milky fluid. At histology the main findings were represented by multiple foci of amorphous calcified material bordered by proliferating macrophages, fibroblasts and multinucleated giant cells, separated by fibrous septa. The following characteristics are worth of note: the presentation in adulthood (late onset); the Caucasian race of the patient; the radiological appearances indicative of hydatid cyst; the aggressive clinicopathologic course, with multifocal involvement, rapid growth, infiltrative pattern. Histologically the TC exhibited the typical pattern of stage II according to Slavin's classification of disease. PMID- 15792375 TI - [Where is telepathology headed?]. PMID- 15792367 TI - Apoptosis and oncosis in acute coronary syndromes: assessment and implications. AB - The rational design of therapeutic interventions for protection of ischemic myocardium from ultimate death requires an understanding of the mechanistic basis of cardiomyocyte (CM) cell death, its timing and the tools for its quantification. Until recently, CM cell death following ischemia and/or reperfusion was considered to involve necrosis or 'accidental cell death' from very early on. Collective evidence over the past decade indicates that early CM cell death after myocardial ischemia and post-ischemic reperfusion involves apoptosis with cell shrinkage and drop-out, and/or oncosis with cell swelling followed by necrosis. This paradigm shift suggests that different approaches for cardioprotection are required. Oncologists, pathologists, anatomists and basic scientists who have studied apoptosis over the last three decades separated physiological apoptosis from inappropriate apoptosis in pathological states. Until recently, cardiologists resisted the concepts of CM apoptosis and regeneration. Cumulative evidence indicating that apoptosis in the heart may occur in different cell types, spread from one cell type to another, and occur in bursts, may have profound implications for therapies aimed at protection of ischemic myocardium by targeting CM apoptosis in acute coronary syndromes. This review focuses on a critique of the methods used for the assessment of CM apoptosis and the implications of CM apoptosis in acute coronary syndromes. PMID- 15792374 TI - Osteosarcoma with rhabdomyosarcomatous component or so-called malignant mesenchymoma of bone. AB - BACKGROUND: Primary malignant mesenchymoma of the bone is a rare neoplasm consisting of two or more unrelated malignant mesenchymal components. The literature reports fewer than 20 cases, most of which were composed of osteosarcoma and liposarcoma. OBSERVATION: We report an exceedingly rare case of primary malignant mesenchymoma of bone composed of rhabdomyosarcoma, osteosarcoma, and a minor chondrosarcoma component, arising in the right proximal humerus of a 15-year-old girl. The rhabdomyosarcomatous component was present in the initial biopsy and persisted in surgical specimen following chemotherapy. CONCLUSION: Effect of chemotherapy is enigmatic since rhabdomyosarcomatous component could appear, persist or disappear after chemotherapy according to literature. PMID- 15792377 TI - Body, mind and spirit. what doctors need to know about the Scottish Health Department's spirituality initiative. PMID- 15792376 TI - Tumoral, quasitumoral and pseudotumoral lesions of the superficial and somatic soft tissue: new entities and new variants of old entities recorded during the last 25 years. Part V: excerpta III. PMID- 15792378 TI - Update on the management of variceal bleeding. PMID- 15792379 TI - Metal-on-metal hip resurfacing. PMID- 15792380 TI - Prospective observational survey of the utilisation of anaesthetists and the outcome following cardiac arrest calls. AB - BACKGROUND AND AIMS: We wanted to determine whether the practice of routinely sending an anaesthetist to cardiac arrests is common within Scotland. We also wished to evaluate the interventions performed by our intensive care anaesthetist when responding to cardiac arrest calls. METHODS: We performed a telephone survey of the 26 Scottish hospitals with an intensive care unit. We conducted a prospective observational survey over a period of six months in one Scottish teaching hospital. Structured interviews with the anaesthetist who responded to the cardiac arrest call were undertaken. RESULTS: Routine attendance of an anaesthetist at cardiac arrests occurs in 25 of the 26 hospitals surveyed. We analysed 68 of 73 arrest calls. In 28 calls (41%) there was no requirement for anaesthetic intervention. In 40 (59%) the anaesthetist intervened. The interventions were for cardiac arrest procedures in 33 cases and ventilatory failure in the remaining 7 cases. One patient survived to hospital discharge: a mortality of 98%. CONCLUSIONS: Patients who remain in cardiac arrest upon the arrival of the anaesthetist have a very high mortality. The practice of routinely sending an anaesthetist to cardiac arrest calls is not justified. PMID- 15792381 TI - Temporal scanner thermometry: a new method of core temperature estimation in ICU patients. AB - BACKGROUND AND AIMS: Temperature measurement is a routine task of patient care, with considerable clinical impact, especially in the ICU. This study was conducted to evaluate the accuracy and variability of the Temporal Artery Thermometer (TAT) in ICU-patients. Therefore, a convenience sample of 57 adult patients, with indwelling pulmonary artery catheters (PAC) in a 40-bed intensive care unit in a university teaching hospital was used. METHODS: The study design was a prospective, descriptive comparative analysis. Body temperature was thereby measured simultaneously with the TAT and the Axillary Thermometer (AT), and was compared with the temperature recording of the PAC. The use of vasoactive medication was recorded. RESULTS AND CONCLUSIONS: Mean temperature of all measurements was: PAC: 37.1 degrees C (SD: 0.87), TAT 37.0 degrees C (SD: 0.68) and axillary thermometer: 36.6 degrees C (SD: 0.94). The measurements of the TAT and the PAC were not significantly different (man differnce: 0.14 degrees C; SD: 0.51; p = 0.33); whereas the measurements of the PAC and the AT differed significantly (mean difference: 0.46 degerees C; SD: 0.39; p < 0.001). Mean diference in PAC versus TAT analyses, between patients with vasopressor therapy (0.12 degrees C; SD: 0.55), and without vasopressor therapy (0.19 degrees C; SD: 0.48) was not statistically significant (p = 0.47). CONCLUSION: We can conclude that the temporal scanner has a relatively good reliability with an acceptable accuracy and variability in patients with normothermia. The results are comparable to those of the AT, but they do not seem to be sufficient to prove any substantial benefit compared to rectal, oral or bladder thermometry. PMID- 15792382 TI - Quality of life in Parkinson's disease: movement disorders clinic vs general medical clinic--a comparative study. AB - OBJECTIVES: to determine the effect of attending a movement disorders (MD) clinic on quality of life (QOL) outcomes for patients with Parkinson's disease (PD). METHODS: Postal questionnaire study of forty-two patients with Parkinson's disease attending either a movement disorders clinic or more conventional general medical clinic were selected consecutively to complete the Parkinson's Disease Quality of Life Questionnaire (PDQL). All patients were diagnosed by a consultant physician with an interest in Parkinson's disease (S.B.R.) and had attended either the movement disorders clinic or the general medical clinic on at least three occasions. Questionnaires were completed independently of the examiners and returned by post. RESULTS: Mean PDQL score was 124.1 [5.16] in the movement disorders clinic and 95.9 [5.86] in the general medical clinic. Analysis of covariance revealed that those subjects attending the MD clinic reported a significantly higher QOL than those subjects in general medical care (F(1,39)= 161.98, P < 0.001). CONCLUSION: These data indicate that the quality of life of respondents attending the movement disorders clinic is significantly higher than those attending the general medical clinic. PMID- 15792383 TI - The Adults with Incapacity (Scotland) Act--Who knows? Who cares? AB - The Adults with Incapacity (Scotland) Act 2000 provides new guidelines on obtaining consent for adults who are incapable of providing informed consent. This article surveys current practice of health workers when obtaining consent from such individuals, and the results are compared to the Incapacity Act and local guidelines. The survey results suggest that practice of the guidelines is variable and often legally inadequate. AIMS: To quantify awareness and practical application of the Adults with Incapacity (Scotland) Act in surgical wards in a central Glasgow teaching hospital, 2 years after the Act's introduction. METHODS: An interviewer led anonymous questionnaire was presented to all degree educated staff encountered on random visits to acute and elective surgical and orthopaedic wards in a split site teaching hospital. RESULTS: 17 out of 50 staff approached (34%) had not heard of the Adults with Incapacity Act. There was a significantly higher level of awareness amongst staff based on acute wards (85%) when compared with those from elective wards (54%) (p < 0.05, chi-square) There was a complete absence offormal training in using the Act and knowledge was sometimes inaccurate or incorrect. Most staff that had heard of the Act could suggest causes of incapacity, but some included physical disability. There was a persistence of the belief that a relative can consentfor an incapable adult patient, and this was expressed by some of the staff despite knowledge of the Act. CONCLUSIONS: This study shows that a significant number of ward staff have no knowledge of the Act; and understanding is variable amongst staff who have heard of it. The Adults with Incapacity (Scotland) Act 2000 is a unique piece of legislation to protect the interests of incapable patients, and doctors treating them; but is of no use if medical and nursing staff are unaware of its existence. PMID- 15792384 TI - Surgical treatment of in-growing toenails performed by senior house officers: are they good enough? AB - BACKGROUND: Although the surgical treatment of in-growing toenails is a common procedure, the success of ingrown toenail surgery is extremely variable and recurrences frequently impair the quality of lift of those who have this condition. In most hospitals this procedure is commonly performed by junior surgical trainees who may have little experience. AIM: We proposed to find out if the success of this procedure is operator-dependent by comparing the results of standard of toenail surgery performed by basic surgical trainees (BSTs) in our hospitals with already published data. METHODS: A retrospective analysis of nail bed ablation surgery performed by BSTs under local anaesthesia over a 15-month period in a district general hospital was conducted. RESULTS: 106 phenol ablations and 46 germinal matrix excisions were prformed. Symptomatic recurrence rates 12 months following the procedure were 5.7% for phenol ablation and 4.3% for germinal matrix excisions. CONCLUSION: Our results are comparable to published data, and we conclude that toenail ablation surgery can be just as successfully performed by junior surgeons after relatively little training in the procedure. PMID- 15792385 TI - Can alcohol intoxication be excluded as the cause of confusion following head injury? AB - OBJECTIVE: To assess whether there is a breath alcohol level (BrAC) below which confusion in the head injured patient should not be attributed solely to the acute effects of alcohol. METHOD: Based in the Accident and Emergency Ward in Glasgow Royal Infirmary, a prospective observational study was carried out over a five month period. Patients admitted to the ward were recruited for the study if they had a primary diagnosis of head injury. The outcome measures recorded and analysed were sequential 2 hrly BrAC readings (mg/L) and Glasgow Coma Scale findings (Eye opening, motor and verbal responses). The relationship between these was investigated, which revealed additional relevant factors affecting level of consciousness. RESULTS: The breath alcohol analyser was found to be a useful non-invasive, quick and easy to use tool. The results obtained were consistent with the expected pattern of reducing BrAC levels over a 6 hour period. Within this group of patients, a poor correlation was found between each of the three responses of the Glasgow Coma Scale and BrAC readings. For those patients who remained confused, when their BrAC reading was less than 1 mg/L, other causes of a lowered level of consciousness were identified. CONCLUSION: Confusion in the head injured patient with a BrAC of less than 1 mg/L, should alert one to the likelihood of causes other than alcohol intoxication. PMID- 15792386 TI - The urinary 5-hydroxyindole acetic acid and plasma nitric oxide levels in irritable bowel syndrome: a preliminary study. AB - BACKGROUND AND AIMS: Postprandial increase of 5-hydroxytryptamine (5-HT) has been implicated in irritable bowel syndrome (IBS). There is evidence that nitric oxide (NO) may act as a mediator of 5-HT-evoked secretions in the colon. Our aim is to investigate the role of urinary 5-hydroxyindole acetic acid (5-HIAA) and plasma NO levels (with diarrhoea) in IBS patients. METHODS: Nineteen (with constipation) IBS patients (group 1), 22 IBS patients (group 2) and 18 healthy controls (group 3) were included in the study. The diagnosis of IBS was made according to the Rome I Criteria. The urine was collected for determination of 5-HIAA and venous blood was collected from each subject for the measurement of plasma NO levels. RESULTS: The levels of urinary 5-HIAA mmol/day and plasma NO mmol/l of group 1 (22.4 +/- 2.2 and 29.4 +/- 2 respectively) were significantly higher than group 3 (14.2 +/- 2.3 and 21.3 +/- 2.1 respectively) (p = 0.036 and p = 0.019 respectively). The NO level of group 1 was also significantly higher than group 2 (21.8 +/- 1.9) (p = 0.021). The 5-HIAA level of group 1 was higher than group 2 (15.2 +/- 2.1) and the difference was marginally significant (p = 0.055). There was no difference between group 2 and group 3 with respect to 5-HIAA and NO levels. CONCLUSIONS: The results of this preliminary study lend support to the involvement of 5-HT in some symptomatology of diarrhoea predominant IBS. Furthermore, NO may be one of the effector mediators of the 5-HT-induced symptoms in these patients. PMID- 15792387 TI - Weight loss and fatigue in an elderly patient. AB - We report a recent clinical case of an elderly patient who presented with weight loss and fatigue and who was diagnosed with hyperthyroidism. Some time later, he was discovered to have underlying oesophageal lymphoma. The latter diagnosis was believed to contribute, at least in part, to his symptoms at the time of initial presentation. The case illustrates the potential difficulties associated with establishing the diagnosis of oesophageal lymphoma. Furthermore, symptoms of weight loss and fatigue are commonly encountered in general medical patients. This case reminded us that even when a diagnosis of hyperthyroidism has been established, these symptoms may be attributable to other pathology. PMID- 15792388 TI - Metastatic carcinoma presenting as cranial nerve palsy. AB - Adult onset, multiple oculo-motor nerve palsies are usually due to neoplasia and trauma. We report two cases whose initial presenting sign was oculo-motor cranial nerve palsy and were later found to have metastatic carcinoma. PMID- 15792389 TI - Primary hyperparathyroidsm, amyloid and multiple myeloma: an unusual association. AB - We report the case of a 76-year-old woman with a diagnosis of Primary Hyperparathyroidsm and Systemic Amyloidosis, in whom subsequent investigations revealed the presence of Multiple Myeloma. We discuss the relationship between these conditions and the implications for management. PMID- 15792390 TI - Hyaline membrane and neonatal radiology--Ian Donald's first venture into imaging research. AB - While he was working at the Royal Postgraduate Medical School, Ian Donald (later Regius Professor of Midwifery, University of Glasgow and a pioneer of diagnostic ultrasound) collaborated with Albert Claireaux and Robert Steiner on histological and radiological studies of hyaline membrane disease. In 1953, Donald and Steiner published thefirst radiological study of a series of cases. The success of this research stimulated Donald's interest in imaging technologies. PMID- 15792391 TI - [Research methods in dentistry 9. Follow-up of permucosal implants in an edentate mandible using panoramic radiography]. AB - Panoramic radiographs are frequently used for routine follow-up of mandibular implants. The objective of this study was to determine whether measurement on a panoramic radiograph of the vertical dimension of the mandible near by an implant, using the known implant length as a reference, is a reliable method. In 11 patients, 2 permucosal implants were placed in the anterior part of the edentulous mandible. During the first year after implantation, 2 panoramic radiographs and 2 sets of standardized oblique lateral cephalometric radiographs were made. Oblique lateral cephalometric radiographs are the golden standard for measuring the vertical dimension of an edentulous mandible. The length of the implants and the vertical dimension of the mandible dorsally to the implants were measured on all oblique lateral cephalometric radiographs. The measured and known implant length were used to calculate the image enlargement factor. This factor was used to calculate the real vertical dimension of the mandible. The same measurement procedures were performed on the panoramic radiographs. Using a paired t-test, the calculated values of the vertical dimensions of the mandibles found on panoramic radiographs were compared with the calculated values found on oblique lateral cephalometric radiographs. No statistically significant differences were found. It was concluded that under the described circumstances, panoramic radiographs can be used for reliable measurement of the vertical dimension of the mandible near by permucosal implants. PMID- 15792392 TI - [Antibiotic prophylaxis for patients with joint prostheses]. AB - In The Netherlands only patients with reduced defence mechanism are considered for antibiotic prophylaxis, and only in case of a total hip replacement. The extend to which the American indications will be applied in Europe, is at this moment unknown. It becomes more and more clear that the risk from providing prophylaxis is greater than the risk of a joint infection. PMID- 15792393 TI - [Guidelines for the treatment of highly anxious dental patients]. AB - Treatment of extreme or pathological forms of dental anxiety requires specific skills. This paper is an attempt to formulate a number of treatment strategies, based on the available literature and effectiveness studies. It is suggested that based upon the assessment information three types of problem areas can be identified, namely, 1. a phobia of specific dental procedures or situations; 2. long overdue requiring acute dental treatment; and 3. intrusive or interfering psychiatric symptoms. In this article these problem situations are reviewed and several appropriate treatment options are discussed. PMID- 15792394 TI - [(Pain) complaints that are "not understood" after implant surgery. Diagnosis with CT/DentaScan]. AB - Combining clinical examination and panoramic, periapical or lateral cephalometric radiographs is generally sufficient in the pre- and post-operative investigation of most implant patients. Sometimes there is a need for more information needing computer tomography (CT) as an additional diagnostic tool. The obtained CT data can be manipulated and reconstructed with software programs such as DentaScan. The resulting cross sections, that can be given in true proportions, are perpendicular to the course of the jaw. Interpretation of these processed images is much easier but not always more accurate than the original scans. Pre operatively, CT/DentaScans can be used not only for the determination of bone quantity and quality, but also for the location of relevant anatomical structures. Post-operatively, the scans can be valuable in the diagnosis of unexplained problems that may be related to the placed implants. Three case reports illustrate the use and indication of CT/DentaScans in solving implant related maxillary pathology or neuralgic pain. PMID- 15792395 TI - Interpretation of absorption rate data for inhaled fluticasone propionate obtained in compartmental pharmacokinetic modeling. AB - OBJECTIVE: Reports characterizing the pharmacokinetics of inhaled fluticasone propionate (FP) using compartmental approaches have suggested that the absorption of FP into the systemic circulation is rapid with a half-life of approximately 10 min. We believe that this is a classical case of misassignment of the pharmacokinetic parameter estimates, a problem often encountered while modeling pharmacokinetic data. The objective of this study was to illustrate and analyze this problem using actual blood level data of FP obtained in 14 healthy subjects. MATERIALS AND METHODS: Serum concentration-time data of FP were obtained from a double-blind, randomized study involving single and multiple twice-daily inhalations of 500 microg via a dry powder device, Diskus. The profiles were fitted using one- and two-compartment pharmacokinetic models with first order absorption. Various permutations of the resulting exponential rate constants were analyzed to determine the combination that was most consistent with the underlying physical process. RESULTS: The two-compartment body model with first order absorption gave excellent fits for the observed FP concentrations after both single and multiple dosing. Even though peak levels were reached relatively early (30 - 90 min) after inhalation, the combination that most appropriately described the underlying process was alpha > Ka > beta, i.e. slow absorption, rapid distribution and slower elimination kinetics. The absorption, distribution and elimination half-lives resulted to be 3.8 h, 9.9 min and 13.6 h, respectively, consistent with the high lipophilicity and sustained dissolution characteristics observed in vitro. CONCLUSIONS: Analysis of FP pharmacokinetics after inhalation represents a classical case of potential misassignment of the exponential rate constants, which if ignored, could lead to erroneous interpretations regarding the underlying process. The study also elucidates the pitfall of using t(max) to calculate absorption rate. PMID- 15792396 TI - Effect of ketoconazole on the pharmacokinetics and safety of telithromycin and clarithromycin in older subjects with renal impairment. AB - OBJECTIVE: The objective of this study was to determine the effect of multiple impairments in drug elimination on the pharmacokinetics and pharmacodynamics (effect on QTc interval), using clarithromycin as a comparator. METHODS: Thirty two subjects aged > or = 60 years with renal impairment who were otherwise medically stable were recruited into this parallel-group study. Following stratification according to creatinine clearance (CL(CR)), subjects were randomized to a five-day treatment with ketoconazole (400 mg once daily) alone, or a five-day treatment with ketoconazole (400 mg once daily) and telithromycin (800 mg once daily) given concomitantly or a five-day treatment with ketoconazole (400 mg once daily) and clarithromycin (500 mg twice daily) given concomitantly. Steady-state pharmacokinetics and safety, including serial electrocardiograms, were assessed. RESULTS: In subjects with CL(CR) 30 - 80 ml/min, the mean maximal telithromycin concentration at steady state (C(max),ss) was 3.6 mg/l and the steady state area under the plasma concentration-time curve from time zero to 24 hours (AUC(0-24 h) ss) was 33.4 mg x h/l. The mean C(max), ss and AUC(0-12 h)ss for clarithromycin were 6.2 mg/l and 56.1 mg x h/l, respectively. The increases in telithromycin C(max) ss and AUC(0-24 h) ss compared to corresponding data for healthy young subjects were 1.6- and 2.7-fold, respectively, whereas corresponding increases for clarithromycin were 2.2- and 3.3-fold, respectively. In the telithromycin plus ketoconazole group deltaQTc values were equal or < 60 ms. All QTc values were equal or < 450 ms in males and equal or < 470 ms in females. CONCLUSIONS: The increase in telithromycin plasma concentrations during ketoconazole-mediated inhibition of CYP3A4 in subjects aged 60 years or older with renal impairment was similar to that for clarithromycin under the same conditions. Telithromycin was well tolerated and produced no clinically significant prolongations in the QTc interval. PMID- 15792397 TI - Pharmacokinetics of scopolamine in serum and subcutaneous adipose tissue in healthy volunteers. AB - OBJECTIVE: The objective was to develop a microdialysis set-up to measure the concentration-time course of scopolamine in the interstitium of subcutaneous adipose tissue. MATERIALS AND METHODS: Six healthy male volunteers were eligible for data analysis. Subjects received 0.5 mg scopolamine as a 15-minute intravenous infusion. Microdialysis samples from interstitial space fluid of subcutaneous adipose tissue and blood samples were taken at predefined intervals over a period of 360 minutes. Scopolamine concentrations were measured by liquid chromatography-tandem mass spectrometry (LC-MS-MS). RESULTS: High inter individual variability was observed in all pharmacokinetic parameters. The mean peak serum concentration (C(max)) of 6.5 +/- 3.9 ng/ml (data in mean +/- SD) was attained after 15 +/- 3 minutes (t(max)), whereas in dialysate, a mean peak concentration of 2.7 +/- 1.7 ng/ml was measured after 27 +/- 8 minutes. The ratio of the area under the concentration versus time curve from 0-360 min for interstitium (AUC(interstitium 0-360 min0) to the AUC for serum (AUC(serum 0-360 min)) was 0.96 +/- 0.7. The elimination half-life of scopolamine was 121 +/- 85 minutes in serum and 166 +/- 117 minutes in dialysate. Values for total clearance and volume of distribution in serum were 99.1 +/- 35.0 1/h and 188 +/- 76 1, respectively. CONCLUSIONS: In the present study, we were able to define a microdialysis set-up, which allows for the measurement of scopolamine concentrations in target tissues. In addition, we demonstrated that the concentrations of scopolamine in subcutaneous adipose tissue resemble closely the concentration-time course in serum of healthy volunteers. PMID- 15792399 TI - Frequency distribution of dextromethorphan O-demethylation in a Greek population. AB - OBJECTIVE: To determine the CYP2D6 phenotype in a Greek population by using dextromethorphan (DM) as a probe drug. METHODS: DM (30 mg) was given orally to 102 unrelated Greek subjects and 8-hour urine samples were collected. Concentrations of DM and its metabolite dextrorphan (DX) were determined using a validated HPLC assay. Metabolic molar ratio (MR) of DM to free DX in log form was used as an in vivo index of metabolic status. RESULTS: The frequency distribution histogram of MR was bimodal. An antimode of 0.25 for the mean log MR was determined using probit analysis. Seven of 102 subjects (6.9%) were poor metabolizers (PMs). CONCLUSION: The PM frequency of CYP2D6 in Greek subjects was similar to other Caucasian populations. PMID- 15792398 TI - Pharmacokinetics of ibuprofen sodium dihydrate and gastrointestinal tolerability of short-term treatment with a novel, rapidly absorbed formulation. AB - OBJECTIVE: This paper describes four studies investigating the dissolution, plasma pharmacokinetics and safety of a novel, fast-acting ibuprofen formulation, ibuprofen sodium dihydrate. MATERIAL AND METHOD: Four separate studies investigated: the in vitro dissolution rates of ibuprofen sodium dihydrate (at pH 1.2, 3.5 and 7.2); the bioavailability of ibuprofen sodium dihydrate (in two pharmacokinetic studies; combined n = 38) compared with conventional ibuprofen, ibuprofen lysinate, ibuprofen arginate and ibuprofen liquagels (all 2 x 200 mg ibuprofen); and the gastroduodenal tolerance of ibuprofen sodium dihydrate and ibuprofen arginate (both 2 x 200 mg ibuprofen t.i.d.) in an endoscopy safety study, where endoscopy was performed at baseline and at the end of each treatment period using a five-point scale to assess the integrity of the gastric and duodenal mucosa. RESULTS: Ibuprofen sodium dihydrate dissolved significantly more rapidly at pH 1.2, 3.5 and 7.2 than conventional ibuprofen, ibuprofen lysinate and ibuprofen liquagels. Ibuprofen sodium dihydrate had similar C(max) to ibuprofen lysinate and ibuprofen liquagels and significantly higher Cmax than conventional ibuprofen (p = 0.002). The mean plasma concentration for ibuprofen sodium dihydrate was significantly higher than for conventional ibuprofen (p = 0.028) 10 minutes post-dose and the t(max) for ibuprofen sodium dihydrate was reached significantly earlier than for conventional ibuprofen (p = 0.018). All three formulations were bioequivalent according to the acceptable boundaries (90% confidence intervals). No statistically significant difference was observed between the ibuprofen formulations in terms of adverse events and specifically with respect to hemorrhagic scores; 41 (46.0%) adverse events (AEs) occurred after administration of ibuprofen sodium dihydrate, and 46 (52.9%) after ibuprofen arginate. One occurrence of an invasive ulcer was observed after administration of ibuprofen arginate. CONCLUSIONS: The new formulation of ibuprofen sodium dihydrate dissolves quickly in vitro, has the same extent of absorption as other fast-acting ibuprofen formulations, and is absorbed into plasma more rapidly than conventional ibuprofen. In addition, the present studies suggest that the tolerability and safety profile of ibuprofen sodium dihydrate is comparable to existing ibuprofen formulations. PMID- 15792400 TI - Assessment of the bioequivalence of two nelfinavir tablet formulations under fed and fasted conditions in healthy subjects. AB - OBJECTIVES: This study was designed to assess the bioequivalence between the commercial 250 mg nelfinavir tablet and the new 625 mg nelfinavir tablet (Roche) which was developed to reduce the daily pill burden for patients from 10 to 4 tablets in a nelfinavir 1250 mg twice daily regimen. METHODS: A total of 52 healthy male subjects were enrolled in this randomized four-period crossover study to receive single oral doses of 1250 mg nelfinavir administered as five commercial 250 mg tablets (reference formulation) and as two new 625 mg tablets (test formulation). Each of the two formulations were taken after an overnight fast and immediately after intake of a standard breakfast (820 kcal) on separate occasions. Blood samples were collected pre-dose and at appropriate intervals after drug administration. Plasma concentrations of nelfinavir and its main metabolite M8 were assayed by a validated LC-MS/ MS assay and the pharmacokinetics of nelfinavir and M8 were derived using standard non compartmental analysis. RESULTS: The primary parameters for bioequivalence testing were the logarithmically transformed AUC(0-inf) and C(max) of nelfinavir taken from 50 subjects who completed all four treatments. Bioequivalence was accepted if the 90% confidence interval (CI) was contained entirely in the equivalence region (80%, 125%). In the fed state, this criterion was met for AUC (effect ratio = 95%; CI = 87%, 103%) and Cmax (effect ratio = 101%; CI = 94%, 109%) and bioequivalence of the two treatments could be concluded. In the fasted state, AUC clearly failed to meet the bioequivalence criteria (effect ratio = 73%; CI = 59%, 90%) and Cmax was borderline outside the lower acceptance region (effect ratio = 97%; CI = 79.6%, 118%). Therefore, bioequivalence could not be concluded under fasted condition. Food increased the systemic exposure to nelfinavir (as reflected by comparison of the logarithmically transformed AUC(0 inf) values under fed and fasted conditions) by six- and eight-fold after dosing with the 250 mg and the 625 mg tablet, respectively. CONCLUSIONS: Bioequivalence of the new 625 mg nelfinavir tablet relative to the commercial 250 mg tablet, at a dose of 1250 mg, was confirmed in the fed state but not under fasted conditions. As nelfinavir is recommended to be taken with food, the new tablet is well-suited to decrease the daily pill burden for patients on a nelfinavir twice daily regimen and to enhance patient's compliance and adherence. PMID- 15792401 TI - Use of health services for disorders of the female reproductive system: United States, 1977-78. AB - Using data from the National Ambulatory Medical Care Survey and the National Hospital Discharge Survey, statistics are presented on visits to office-based physicians for care and treatment of female reproductive disorders and on patients discharged from short-stay hospitals with related surgery. Visits are described in terms of patient, physician, and clinical characteristics. Breast and gynecological surgery is shown in terms of associated diagnoses and is charted for 1970-78. PMID- 15792402 TI - Self-assembled peptide amphiphile nanofibers conjugated to MRI contrast agents. AB - Self-assembled peptide amphiphile nanofibers have been investigated for their potential use as in vivo scaffolds for tissue engineering and drug delivery applications. We report here the synthesis of magnetic resonance (MR) active peptide amphiphile molecules that self-assemble into spherical and fiber-like nanostructures, enhancing T(1) relaxation time. This new class of MR contrast agents can potentially be used to combine high-resolution three-dimensional MR fate mapping of tissue-engineered scaffolds with targeting of specific cellular receptors. PMID- 15792403 TI - High-density silver nanoparticle film with temperature-controllable interparticle spacing for a tunable surface enhanced Raman scattering substrate. AB - The formation of high-density silver nanoparticles and a novel method to precisely control the spacing between nanoparticles by temperature are demonstrated for a tunable surface enhanced Raman scattering substrates. The high density nanoparticle thin film is accomplished by self-assembling through the Langmuir-Blodgett (LB) technique on a water surface and transferring the particle monolayer to a temperature-responsive polymer membrane. The temperature responsive polymer membrane allows producing a dynamic surface enhanced Raman scattering substrate. The plasmon peak of the silver nanoparticle film red shifts up to 110 nm with increasing temperature. The high-density particle film serves as an excellent substrate for surface-enhanced Raman spectroscopy (SERS), and the scattering signal enhancement factor can be dynamically tuned by the thermally activated SERS substrate. The SERS spectra of Rhodamine 6G on a high-density silver particle film at various temperatures is characterized to demonstrate the tunable plasmon coupling between high-density nanoparticles. PMID- 15792404 TI - Rapid and reproducible fabrication of carbon nanotube AFM probes by dielectrophoresis. PMID- 15792405 TI - Dynamic shadow mask technique: a universal tool for nanoscience. AB - A comprehensive instrument, designed for fabricating nanostructures by evaporation through a dynamic shadow mask in ultrahigh vacuum, is described. The versatility and performance of the instrument is demonstrated through a series of examples, allowing for applications that are impossible to achieve with traditional nanopatterning methods. Clean nanostructures or entire devices made of different materials and on various substrates can be fabricated. The technique is compatible with fundamental surface science and can be easily interfaced with other fabrication and characterization techniques. PMID- 15792406 TI - Biocatalytic growth of Au nanoparticles: from mechanistic aspects to biosensors design. AB - The H(2)O(2)-mediated enlargement of Au nanoparticles (NPs) and the growth mechanism are described. In addition to the deposition of gold on the NP faces, the formation of nanocrystalline clusters at the intersection of the faces is observed. The detachment of the latter nanoclusters provides additional seeds for the deposition of gold. The biocatalyzed generation of H(2)O(2) in the presence of O(2)/glucose and glucose oxidase enabled the development of an optical biosensor for glucose. PMID- 15792407 TI - Nanoscale operation of a living cell using an atomic force microscope with a nanoneedle. AB - We have developed a tool for performing surgical operations on living cells at nanoscale resolution using atomic force microscopy (AFM) and a modified AFM tip. The AFM tips are sharpened to ultrathin needles of 200-300 nm in diameter using focused ion beam etching. Force-distance curves obtained by AFM using the needles indicated that the needles penetrated the cell membrane following indentation to a depth of 1-2 microm. The force increase during the indentation process was found to be consistent with application of the Hertz model. A three-dimensional image generated by laser scanning confocal microscopy directly revealed that the needle penetrated both the cellular and nuclear membranes to reach the nucleus. This technique enables the extended application of AFM to analyses and surgery of living cells. PMID- 15792408 TI - Edge-spreading lithography: use of patterned photoresist structures to direct the spreading of alkanethiols on gold. AB - Edge-spreading lithography (ESL) has been extended to fabricate gold structures with different geometries and feature sizes on silicon substrates. In the present variant of ESL, we used photoresist structures patterned on a thin film of gold to transport alkanethiol molecules from an elastomeric stamp to the gold surface where they formed a self-assembled monolayer (SAM) along the edge of each resist feature. The emerging SAM could expand laterally on the gold via reactive spreading, during which the footprints of the resist structures were enlarged in the same fashion. Upon removal of the photoresist, selective etching of gold revealed those regions that were protected by the SAM, yielding accurate outlines of the resist features on the substrate. The width of resultant gold structures was determined by the distance over which the monolayer expanded during spreading, and could be conveniently controlled well below 200 nm by varying the contact time and/or the concentration of alkanethiol in the ink. PMID- 15792409 TI - Dual-luminophore-doped silica nanoparticles for multiplexed signaling. AB - We have synthesized dual-luminophore-doped silica nanoparticles for multiplexed signaling in bioanalysis. Two luminophores, Tris(2,2' bipyridyl)osmium(II)bis(hexafluorophosphate) (OsBpy) and Tris(2,2' bipyridyl)dichlororuthenium(II)hexahydrate (RuBpy), were simultaneously entrapped inside silica nanoparticles at precisely controlled ratios, with desirable sizes and required surface functionality. Single-wavelength excitation with dual emission endows the nanoparticles with optical encoding capability for rapid and high-throughput multiplexed detection. The nanoparticles can be prepared with sizes ranging from a few nanometers to a few hundred nanometers, with specific ratios of luminescence intensities at two well-resolved wavelengths and with excellent reproducibility. These nanoparticles also possess unique properties of high signal amplification, excellent photostability, and easy surface bioconjugation for highly sensitive measurements when used as signaling markers. A simplified ligand binding system using avidin-biotin and an application extension to immunoassays have been explored, demonstrating the potential use of these easily obtainable bioconjugated nanoparticles for multiplexed signaling and bioassays. PMID- 15792410 TI - Amphiphilic network as nanoreactor for enzymes in organic solvents. AB - Enzymes are powerful biocatalysts that work naturally in water but are also active in organic solvents. Here, we present a nanophase-separated amphiphilic network, where an enzyme is entrapped into its hydrophilic domains. A substrate that diffuses into the other, hydrophobic, phase of such a network can access the biocatalyst via the extremely large interface. Entrapped horseradish peroxidase and chloroperoxidase showed dramatically increased activity and operational stability compared to the native enzymes. PMID- 15792411 TI - Composite organic-inorganic nanoparticles (COINs) with chemically encoded optical signatures. AB - To obtain a coding system for multiplex detection, we have developed a method to synthesize a new type of nanomaterial called composite organic-inorganic nanoparticles (COINs). The method allows the incorporation of a broad range of organic compounds into COINs to produce surface enhanced Raman scattering (SERS) like spectra that are richer in variety than fluorescence-based signatures. Preliminary data suggest that COINs can be used as Raman tags for multiplex and ultrasensitive detection of biomolecules. PMID- 15792412 TI - Tuning the bipolar conductance of an alkali-doped C(60) layer sandwiched between two tunneling barriers. AB - Resonant tunneling through a C(60) monolayer doped with single Na, K, Rb, and Cs atoms was measured between the tip of a scanning tunneling microscope and a NiAl(110) substrate. By supporting the monolayer on a thin aluminum oxide film grown on the substrate, a double barrier tunnel junction is formed, consisting of the vacuum and oxide. This geometry enables conductance through an electronic state of the alkali-C(60) complex at both positive and negative sample bias. The positions of the conductance peaks can be varied by tuning the vacuum barrier. An opposite variation is found for Na and K as compared to Rb and Cs, suggesting the influence of bonding on nanoscale transport. PMID- 15792413 TI - Effect of local environment on molecular conduction: isolated molecule versus self-assembled monolayer. AB - Developing a fundamental understanding of molecular conduction in different device environments is essential to the advance of molecular electronics. We show through a quantitative comparison of two types of junctions with the same molecule - one based on an isolated individual molecule and the other on a self assembled monolayer - that intrinsic differences in the conduction per molecule as large as several orders of magnitude can exist simply as a function of the presence or absence of neighboring molecules. This behavior can be understood on the basis of thermal and electrostatic effects that depend critically on the local molecular environment. These results will help to unify data obtained from disparate device structures and to provide an improved basis for designing future molecular electronic devices. PMID- 15792414 TI - Nanoscale conjugated-polymer light-emitting diodes. AB - We use e-beam lithography to pattern an indium tin oxide (ITO) electrode to create arrays of conjugated-polymer LEDs, each of which has a hole-injecting contact limited to 100 nm in diameter. Using optical microscopy, we estimate that the electroluminescence from a 100 nm diameter LED comes from a region characterized by a diameter of approximately 170 nm. This apparent broadening occurs due to current spreading within a PEDOT:PSS layer which was included to aid hole injection. PMID- 15792415 TI - Probing the conformation of physisorbed molecules at the atomic scale using STM manipulation. AB - We use scanning tunneling microscope (STM) manipulation and density functional theory calculation to investigate the structural properties of individual sexiphenyl molecules physisorbed on a Ag(111) surface at 6 K. The molecule surface atomic registry is precisely determined by using atomic markers and a sexiphenyl functionalized tip. The calculations confirm the alternating twist of the sexiphenyl pi-rings on Ag(111). The pi-ring torsional angle, 11.4 degrees, is directly determined from the geometry of STM manipulation. This innovative experiment opens up a novel application of STM manipulation to probe the properties of "physisorbed" species on surfaces at the atomic level. PMID- 15792416 TI - Two-dimensional self-assembly into multicomponent hydrogen-bonded nanostructures. AB - By means of scanning tunneling microscopy, we have explored the two-dimensional self-assembly of functional bicomponent hydrogen-bonding dye systems, leading to well-defined patterns, different from those of the individual components, and providing design rules to immobilize multicomponent systems at the liquid-solid interface. PMID- 15792417 TI - Directed spatial organization of zinc oxide nanorods. AB - The ability to precisely place nanomaterials at predetermined locations is necessary for realizing applications using these new materials. Using an organic template, we demonstrate directed growth of zinc oxide (ZnO) nanorods on silver films from aqueous solution. Spatial organization of ZnO nanorods in prescribed arbitrary patterns was achieved, with unprecedented control in selectivity, crystal orientation, and nucleation density. Surprisingly, we found that caboxylate endgroups of omega-alkanethiol molecules strongly inhibit ZnO nucleation. The mechanism for this observed selectivity is discussed. PMID- 15792418 TI - Quantum growth of magnetic nanoplatelets of Co on Si with high blocking temperature. AB - Self-organized Co nanoplatelets with a singular height, quantized lateral sizes, and unique shape and orientation have been fabricated on a template consisting of ordered Al nanocluster arrays on Si(111)-7 x 7 surfaces. Despite their small volume (a few nm(3)), these nanomagnets exhibit an unusually high blocking temperature (>100 K). The perpendicular direction for easy magnetization, the high blocking temperature, the size tunability, and the epitaxial growth on Si substrates make these nanomagnets important for applications in information technology. PMID- 15792419 TI - Conductive atomic force microscope nanopatterning of hydrogen-passivated silicon in inert organic solvents. AB - Ambient liquid phase atomic force microscope (AFM) techniques for nanopatterning organic molecules on silicon through direct Si-C bonds rely on reactions that are in direct competition with spurious oxidation. We study the effectiveness of an inert hydrophobic organic solvent at suppressing oxidation of hydrogen-passivated silicon under ambient conditions. Nanometer scale features were fabricated on an H:Si(111) substrate using a conductive AFM in hexadecane. The patterned features show chemical and kinetic behavior consistent with field induced oxidation (FIO) in air. The mechanism for FIO in hexadecane is discussed. PMID- 15792420 TI - Electrostatic trapping of double-stranded DNA by using cadmium hydroxide nanostrands. AB - Cadmium ions (Cd(2+)) spontaneously grew into cadmium hydroxide nanostrands at the proper pH range in water, and the nanostrands efficiently captured negatively charged DNA. The DNA chain adsorbed parallel to the nanostrand due to the numerous positive charges on the extremely long and thin structure of 1.9 nm, giving weakly gelled white precipitates. By the filtration, more than 95% of DNA was separated from the dilute solution with a concentration less than 40 ng/mL. The nanostrands quickly disappeared after adding aqueous EDTA, and DNA was released quantitatively. These characteristics gave a novel separation technique for short DNA fragments without the use of organic solvents. PMID- 15792421 TI - Thrombogenicity and blood coagulation of a microcatheter prepared from carbon nanotube-nylon-based composite. AB - Here we describe the fabrication of nanocomposite-based microcatheter through melt extrusion utilizing high-purity carbon nanotubes as filler and nylon as matrix. Homogeneous dispersion of carbon nanotubes in nylon can provide novel microcatheters with enhanced mechanical properties (easier to handle) and extremely reduced thrombogenicity or antithrombotic property. The present microcatheter with high performance is highly promising for medical applications. PMID- 15792422 TI - Alternative nucleic acid analogues for programmable assembly: hybridization of LNA to PNA. AB - Complementary locked nucleic acid (LNA) and peptide nucleic acid (PNA) hexamers bind to each other with significantly higher affinity than each binds to DNA, and with far greater affinity than DNA binds to complementary DNA. The hybridization is highly specific with a single mismatch causing decreases in T(m) values ranging from 12 (G/T) to 30 degrees C (A/A). Importantly, the hybridization of an LNA oligomer to a PNA oligomer is unaffected by the ionic strength of the buffer. These properties make the LNA/PNA pair an attractive candidate as a replacement for DNA in programmable assembly. PMID- 15792423 TI - Bright and stable core-shell fluorescent silica nanoparticles. AB - A class of highly fluorescent and photostable core-shell nanoparticles from a modified Stober synthesis in the size range of 20-30 nm is described. These nanoparticles are monodisperse in solution, 20 times brighter, and more photostable than their constituent fluorophore, and are amenable to specific labeling of biological macromolecules for bioimaging experiments. The photophysical characteristics of the encapsulated fluorophores differ from their solution properties. This raises the possibility of tuning nanoparticle structure toward enhanced radiative properties, making them an attractive material platform for a diverse range of applications. PMID- 15792424 TI - Nanophotonic crescent moon structures with sharp edge for ultrasensitive biomolecular detection by local electromagnetic field enhancement effect. AB - We present novel gold nanophotonic crescent moon structures with a sub-10 nm sharp edge, which can enhance local electromagnetic field at the edge area. The formation of unconventional nanophotonic crescent moon structure is accomplished by using a sacrificial nanosphere template and conventional thin film deposition method, which allows an effective batch nanofabrication and precise controls of nanostructure shapes. Unique multiple scattering peaks are observed in a single gold nanocrescent moon with dark-field white light illumination. A 785 nm near infrared (NIR) diode laser was used as the excitation source to induce the amplified scattering field on the sharp edge of the single gold nanocrescent moon. The Raman scattering spectrum of Rhodamine 6G molecules adsorbed on the single gold nanocrescent moon are characterized, and the Raman enhancement factor of single gold nanocrescent moon is estimated larger than 10(10), which suggests the potential applications of gold nanocrescent moons in ultrasensitive biomolecular detection and cellular imaging using surface enhanced Raman spectroscopy. PMID- 15792425 TI - Hysteresis, switching, and negative differential resistance in molecular junctions: a polaron model. AB - Within a simple mean-field model (self-consistent Hartree approximation) we discuss the possibility of polaron formation on a molecular wire as a mechanism for negative differential resistance (NDR), switching, and/or hysteresis in the I V characteristic of molecular junctions. This mechanism differs from earlier proposed mechanisms of charging and conformational change. The polaron model captures the essential physics and provides qualitative correspondence with experimental data. The importance of active redox centers in the molecule is indicated. PMID- 15792426 TI - Electrochemical ostwald ripening of colloidal ag particles on conductive substrates. AB - Thermally evaporated silver nanoparticles on conducting substrates spontaneously evolve in size when immersed in pure water. The process was studied using scanning electron microscopy (SEM), energy-dispersive X-ray analysis (EDX), and optical absorption spectroscopy. The particles are proposed to reform through an electrochemical Ostwald ripening mechanism driven by the size dependence of the work function and standard electrode potential. We also discuss prior literature experiments where this process appears to occur. Our results show the sensitivity of the electrochemical properties of metallic nanoparticles at relatively large sizes (approximately 50 nm). PMID- 15792427 TI - Individual single-walled carbon nanotubes as nanoelectrodes for electrochemistry. AB - We demonstrate the use of individual single-walled carbon nanotubes (SWNTs) as nanoelectrodes for electrochemistry. SWNTs were contacted by nanolithography, and cyclic voltammetry was performed in aqueous solutions. Interestingly, metallic and semiconducting SWNTs yielded similar steady-state voltammetric curves. We clarify this behavior through a model that considers the electronic structure of the SWNTs. Interfacial electron transfer to the SWNTs is observed to be very fast but can nonetheless be resolved due to the nanometer critical dimension of SWNTs. These studies demonstrate the potential of using a SWNT as a model carbon nanoelectrode for electrochemistry. PMID- 15792428 TI - Patterning microsphere surfaces by templating colloidal crystals. AB - By using the upper single or double layers in colloidal crystals as masks during Au vapor deposition, various Au patterns have been successfully constructed on the surfaces of the lower spheres. The dimension and geometry of the Au patterns obtained are dependent on the orientation of the colloidal crystal templates. Our patterning procedure is independent of the curvature and chemical composition of the surfaces, which definitely pave a promising way to pattern highly curved surfaces. PMID- 15792429 TI - Sub-20 nm short channel carbon nanotube transistors. AB - Carbon nanotube field-effect transistors with sub-20 nm long channels and on/off current ratios of >10(6) are demonstrated. Individual single-walled carbon nanotubes with diameters ranging from 0.7 to 1.1 nm grown from structured catalytic islands using chemical vapor deposition at 700 degrees C form the channels. Electron beam lithography and a combination of HSQ, calix[6]arene, and PMMA e-beam resists were used to structure the short channels and source and drain regions. The nanotube transistors display on-currents in excess of 15 microA for drain-source biases of only 0.4 V. PMID- 15792430 TI - Synthesis of peapods using substrate-grown SWNTs and DWNTs: an enabling step toward peapod devices. AB - We report the successful synthesis of nanoscale peapods from single-walled and double-walled nanotubes grown by chemical vapor deposition (CVD) on substrates with windows etched into free-standing silicon nitride membranes. CVD-grown nanotubes were oxidized in air, then filled with C(60) molecules from the vapor phase. Observed variation in nanotube oxidation and C(60) packing with nanotube diameter agreed with theoretical expectations. Windowed samples provide several important advantages for property measurements of peapods and other nanomaterials. Individual nanostructures can be followed through processing steps, and a single nanostructure can be inspected by high-resolution TEM and subsequently contacted with nanoscale electrodes using electron beam lithography. PMID- 15792432 TI - Controlled multistep purification of single-walled carbon nanotubes. AB - A controlled and scalable multistep purification method has been developed to remove iron impurity and nonnanotube carbon materials from raw single-walled carbon nanotubes (SWNTs) produced in the HiPco (high-pressure CO) process. In this study, iron nanoparticles, coated by carbon, are exposed and oxidized by multiple step oxidation at increasing temperatures. To avoid catalytic oxidation by iron oxide of carbon nanotubes, the exposed and oxidized iron oxide is deactivated by reaction with C(2)H(2)F(4) or SF(6). The iron fluorides are removed by a Soxhlet extraction with a 6 M HCl solution. The purity and quality of each sample were determined by thermogravimetric analysis (TGA), Raman spectrometry, ultraviolet-visible-near-IR (UV-vis-near-IR) spectrometry, fluorescence spectrometry, and transmission electron microscope (TEM) spectroscopy. The purity and yield of SWNTs are improved due to reduced catalytic activity of the iron oxide. Greater iron oxide removal also resulted from oxidation at higher temperatures. PMID- 15792431 TI - Biological behavior of hat-stacked carbon nanofibers in the subcutaneous tissue in rats. AB - The tissue response to hat-stacked carbon nanofibers (H-CNFs) was evaluated. H CNFs were implanted in the subcutaneous tissue of rats. Histological and ultrastructural investigations were carried out by transmission electron microscopy. Although many macrophages and foreign body giant cells were seen around H-CNFs, no severe inflammatory response such as necrosis was observed. Some H-CNFs were observed in lysosomal vacuoles of phagocytes. These results showed that H-CNFs were not strong prophlogistic substances and were englobed in vivo. PMID- 15792433 TI - Designing van der Waals Forces between Nanocolloids. AB - van der Waals (VDW) dispersion forces are often calculated between colloidal particles by combining the Dzyaloshinskii-Lifshitz-Pitaevskii (DLP) theory with the Derjaguin approximation; however, several limitations prevent using this method for nanocolloids. Here we use the Axilrod-Teller-Muto 3-body formulation to predict VDW forces between spherical, cubic, and core-shell nanoparticles in a vacuum. Results suggest heuristics for "designing" nanocolloids to have improved stability. PMID- 15792434 TI - Self-assembly of metallic nanowires from aqueous solution. AB - Conducting nanowires have been found to self-assemble between microelectrodes from palladium acetate solution on applying an alternating electric field of relatively high intensity and frequency. The wires grew spontaneously along the direction of the electric field and have high uniformity and conductivity. The finding challenges current metallic nanowire methodologies and could accelerate achievement of the direct assembly of nanoscale electronic structures and systems. PMID- 15792435 TI - Cross-linked polymer replica of a nanoimprint mold at 30 nm half-pitch. AB - This letter reports the demonstration of a photocurable polymer process for replicating the master mold for nanoimprint lithography. The cross-linked polymer mold was fabricated directly with high fidelity from a master by imprinting and photocuring a low viscosity liquid prepolymer film spun onto a substrate. The surface of the cross-linked polymer mold can be treated using an O(2) plasma, and then vapor primed with a low surface energy mold release layer for repeatable imprinting. The imprinting results demonstrated that the cross-linked polymer mold could be faithfully used for both thermal and photocurable nanoimprint lithography. PMID- 15792437 TI - Composite nanowire-based probes for magnetic resonance force microscopy. AB - We present a nanowire-based methodology for the fabrication of ultrahigh sensitivity and resolution probes for atomic resolution magnetic resonance force microscopy (MRFM). The fabrication technique combines electrochemical deposition of multifunctional metals into nanoporous polycarbonate membranes and chemically selective electroless deposition of optical nanoreflector onto the nanowire. The completed composite nanowire structure contains all the required elements for an ultrahigh sensitivity and resolution MRFM sensor with (a) a magnetic nanowire segment providing atomic resolution magnetic field imaging gradients as well as large force gradients for high sensitivity, (b) a noble metal enhanced nanowire segment providing efficient scattering cross-section from a sub-wavelength source for optical readout of nanowire vibration, and (c) a nonmagnetic/nonplasmonic nanowire segment providing the cantilever structure for mechanical detection of magnetic resonance. PMID- 15792436 TI - Novel electrochemical biosensing platform using self-assembled peptide nanotubes. AB - Here we describe a novel electrochemical biosensing platform based on biocompatible, well-ordered, self-assembled diphenylalanine peptide nanotubes. Voltammetric and time-based amperometric techniques were applied to demonstrate the ability of the peptide nanotubes to improve the electrochemical parameters of graphite electrodes. The findings clearly show that this novel class of peptide nanotubes provides an attractive component for future electroanalytical devices. PMID- 15792438 TI - Enhanced photocleavage of water using titania nanotube arrays. AB - In this study highly ordered titania nanotube arrays of variable wall thickness are used to photocleave water under ultraviolet irradiation. We demonstrate that the wall thickness and length of the nanotubes can be controlled via anodization bath temperature. We find that the nanotube wall thickness is a key parameter influencing the magnitude of the photoanodic response and the overall efficiency of the water-splitting reaction. For 22 nm inner pore diameter nanotube arrays, those fabricated in a 5 degrees C anodization bath, 224 nm length and 34 nm wall thickness produced a photoanodic response that was thrice that of a nanotube array fabricated in a 50 degrees C anodization bath, 120 nm length and 9 nm wall thickness. At high anodic polarization, above 1 V, the quantum efficiency under 337 nm illumination was greater than 90%. For the 5 degrees C anodization bath samples (22 nm pore-diameter, 34 nm wall thickness), upon 320-400 nm illumination at an intensity of 100 mW/cm(2), hydrogen gas was generated at the power-time normalized rate of 960 micromol/h W (24 mL/h W) at an overall conversion efficiency of 6.8%. To the best of our knowledge, this hydrogen generation rate is the highest reported for a titania-based photoelectrochemical cell. PMID- 15792439 TI - Vacancy formation process in carbon nanotubes: first-principles approach. AB - The electronic and structural properties of a single-walled carbon nanotube (SWNT) under mechanical deformation are studied using first-principles calculations based on the density functional theory. A force is applied over one particular C-atom with enough strength to break the chemical bonds between the atom and its nearest neighbors, leading to a final configuration represented by one tube with a vacancy and an isolated C-atom inside the tube. Our investigation demonstrates that there is a tendency that the first bond to break is the one most parallel possible to the tube axis and, after, the remaining two other bonds are broken. The analysis of the electronic charge densities, just before and after the bonds breaking, helps to elucidate how the vacancy is formed on an atom by-atom basis. In particular, for tubes with a diameter around 11 angstroms, it is shown that the chemical bonds start to break only when the externally applied force is of the order of 14 nN and it is independent of the chirality. The formation energies for the vacancies created using this process are almost independent of the chirality, otherwise the bonds broken and the reconstruction are dependent. PMID- 15792441 TI - Single-crystal 55Mn NMR spectra of two Mn12 single-molecule magnets. AB - The initial application is reported of single-crystal 55Mn NMR spectroscopy, and associated orientation dependence studies, to single-molecule magnets (SMMs). The studies were performed on two members of the Mn12 family of SMMs, [Mn12O12(O2CMe)16(H2O)4].2MeCO2H.4H2O (Mn12-Ac) and [Mn12O12(O2CCH2Br)16(H2O)4].4CH)Cl) (Mn12-BrAc). Single-crystal spectra give a dramatic improvement in the spectral resolution over oriented powder spectra, allowing the clear observation of quadrupolar splittings, the determination of quadrupole coupling parameters (e2qQ), and an assessment of the symmetry-lowering perturbation of the core of Mn12-Ac by hydrogen-bonding interactions with lattice solvate molecules of crystallization. The results emphasize the utility of single crystal NMR studies to probe the cores of these magnetic molecules. PMID- 15792442 TI - A Schiff base expanded porphyrin macrocycle that acts as a versatile binucleating ligand for late first-row transition metals. AB - The coordination chemistry of the Schiff base polypyrrolic octaaza macrocycle 1 toward late first-row transition metals was investigated. Binuclear complexes with the divalent cations Ni(II), Cu(II), and Zn(II) and with the monovalent cation Cu(I) were prepared and characterized. Air oxidation of the Cu(I) ions in the latter complex to their divalent oxidation state resulted in a change in the coordination mode relative to the macrocycle. PMID- 15792443 TI - Inclusion of ferrocene in a cyclodextrin-functionalized layered metal hydroxide: a new organometallic--organic-LDH nanohybrid. AB - Cyclodextrin cavities have been grafted into a layered metal hydroxide to create hydrophobic nanopockets within the galleries of the inorganic solid. Neutral ferrocene molecules can be included within the grafted cavities by partitioning from a polar solvent to generate a new organometallic-organic-inorganic hybrid. The included ferrocene has been characterized by electronic and Raman spectroscopy. The capability of the cyclodextrin-functionalized solid to separate hydrophobic and hydrophilic derivatives of ferrocene is demonstrated. PMID- 15792444 TI - A tris(mu-hydroxy)tricopper(II) complex as a model of the native intermediate in laccase and its relationship to a binuclear analogue. AB - The reaction of a copper(I) complex with a sterically demanding secondary diamine ligand and O2 yields a tris(mu-hydroxy)tricopper(II) complex. This complex is a structural model of the proposed native intermediate in multicopper oxidases, with interesting structural, magnetic, and solution properties. PMID- 15792446 TI - 2-Phosphinophenolate complexes: formation and crystal structure of a novel trinuclear mu-O nickel(II)-tris(P(intersection)O-chelate). AB - A novel linear trinuclear mu-O-bridging 2-phosphinophenolate nickel(II) complex with fac-tris(P(intersection)O- chelates) in the terminal positions and the three oxygen atoms each facing the central nickel(II) cation was synthesized and structurally characterized by X-ray crystallography. To the best of our knowledge, this is the first example of an octahedral Ni(II) tris(P(intersection)O- chelate). PMID- 15792445 TI - beta-Hematin (hemozoin) mediated decompostion of polyunsaturated fatty acids to 4 hydroxy-2-nonenal. AB - beta-Hematin is an important heme metabolite of malarial infection. Its role as an agent mediating the formation of the reactive electrophile 4-hydroxynonenal (HNE) from polyunsaturated fatty acids was investigated. In vitro formation of HNE was found to be facilitated by the presence of hemozoin in a concentration dependent fashion. The reactivity of HNE derived from reaction with beta-hematin was confirmed through its ability to form protein adducts on myoglobin. PMID- 15792447 TI - Selected-control synthesis of metal phosphonate nanoparticles and nanorods. AB - By surfactant-assisted methods, nanoscale Co(O3PC6H5).H2O species of different morphologies, namely, nanoparticles and nanorods, have been successfully synthesized and characterized. Upon removal of the organic part of the compound, peculiar Co2P2O7 porous nanorods formed. PMID- 15792448 TI - Tritopic (cascade) and ditopic complexes of halides with an azacryptand. AB - Structural aspects of the binding of halides in the octaaza cryptand (1,4,11,14,17,24,29,36-octa-azapentacyclo[12.12.12.2(6,9).2(19,22).2(31,34)] tetratetraconta-6(43),7,9(44),19(41),20,22(42),31(39),32,34(40)-nonaene, N(CH2CH2NHCH2-p-xylyl-CH2NHCH2CH2)3N), L1, were examined for fluoride, chloride, and bromide. Crystallographic results for two different fluoride complexes indicated cascade-like coordination, with two fluoride ions inside the tren-based cavity, bridged by a water molecule. In the two different chloride structures, a single chloride and a water molecule occupied the cavity. The bromide structure contained two crystallographically independent cationic cryptands. Unit A consisted of a bromide on one side of the cavity and three disordered water molecules situated between the cryptand arms on the other side. Unit B also had a bromide inside the cavity at one side, but a single molecule of water was centered at the other side of the cavity. Association constants for the three ions, determined by NMR titrations in aqueous solution at pH 5, revealed log Ka=3.15(5), 3.37(3), and 3.34(4) for fluoride, chloride, and bromide, respectively. PMID- 15792450 TI - Reactions of laser-ablated uranium atoms with H2O in excess argon: a matrix infrared and relativistic DFT investigation of uranium oxyhydrides. AB - Laser-ablated U atoms react with H2O during condensation in excess argon. Infrared absorptions at 1416.3, 1377.1, and 859.4 cm(-1) are assigned to symmetric H-U-H, antisymmetric H-U-H, and U=O stretching vibrations of the primary reaction product H(2)UO. Uranium monoxide, UO, also formed in the reaction, inserts into H2O to produce HUO(OH), which absorbs at 1370.5, 834.3, and 575.7 cm(-1). The HUO(OH) uranium(IV) product undergoes ultraviolet photoisomerization to a more stable H2UO2 uranium(VI) molecule, which absorbs at 1406.4 and 885.9 cm(-1). Several of these species, particularly H2UO2, appear to form weak Ar-coordinated complexes. The predicted vibrational frequencies, relative absorption intensities, and isotopic shifts from relativistic DFT calculations are in good agreement with observed spectra, which further supports the identification of novel uranium oxyhydrides from matrix infrared spectra. PMID- 15792449 TI - Oxidations of NADH analogues by cis-[RuIV(bpy)2(py)(O)]2+ occur by hydrogen-atom transfer rather than by hydride transfer. AB - Oxidations of the NADH analogues 10-methyl-9,10-dihydroacridine (AcrH2) and N benzyl 1,4-dihydronicotinamide (BNAH) by cis-[RuIV(bpy)2(py)(O)]2+ (RuIVO2+) have been studied to probe the preferences for hydrogen-atom transfer vs hydride transfer mechanisms for the C-H bond oxidation. 1H NMR spectra of completed reactions of AcrH2 and RuIVO2+, after more than approximately 20 min, reveal the predominant products to be 10-methylacridone (AcrO) and cis [RuII(bpy)2(py)(MeCN)]2+. Over the first few seconds of the reaction, however, as monitored by stopped-flow optical spectroscopy, the 10-methylacridinium cation (AcrH+) is observed. AcrH+ is the product of net hydride removal from AcrH2, but hydride transfer cannot be the dominant pathway because AcrH+ is formed in only 40-50% yield and its subsequent oxidation to AcrO is relatively slow. Kinetic studies show that the reaction is first order in both RuIVO2+ and AcrH2, with k = (5.7 +/- 0.3) x 10(3) M(-1) s(-1) at 25 degrees C, DeltaH(double dagger) = 5.3 +/ 0.3 kcal mol(-1) and DeltaS(double dagger) = -23 +/- 1 cal mol(-1) K(-1). A large kinetic isotope effect is observed, kAcrH2/kAcrD2 = 12 +/- 1. The kinetics of this reaction are significantly affected by O2. The rate constants for the oxidations of AcrH2 and BNAH correlate well with those for a series of hydrocarbon C-H bond oxidations by RuIVO2+. The data indicate a mechanism of initial hydrogen-atom abstraction. The acridinyl radical, AcrH*, then rapidly reacts by electron transfer (to give AcrH+) or by C-O bond formation (leading to AcrO). Thermochemical analyses show that H* and H- transfer from AcrH2 to RuIVO2+ are comparably exoergic: DeltaG degrees = -10 +/- 2 kcal mol(-1) (H*) and -6 +/- 5 kcal mol(-1) (H-). That a hydrogen-atom transfer is preferred kinetically suggests that this mechanism has an equal or lower intrinsic barrier than a hydride transfer pathway. PMID- 15792451 TI - Synthesis, crystal structures, and magnetism of cobalt coordination polymers based on dicyanamide and pyrazine-dioxide derivatives. AB - Three coordination polymers of Co(II) with dicyanamide (dca) were obtained by adding coligands of 2,5-dimethylpyrazine-dioxide (2,5-dmpdo), 2,3,5 trimethylpyrazine-dioxide (2,3,5-tmpdo), or 2,3,5,6-tetramethylpyrazine-dioxide (2,3,5,6-tmpdo) to the binary system of Co-dca. Co2(dca)(4)(2,5-dmpdo)2 (1) crystallizes in the triclinic space group P with a=7.4962(2), b=9.0364(2), c = 10.4783(4) A, alpha = 72.567(1), beta = 72.557(1), gamma = 68.814(2) degrees , V = 616.61(3) A(3), Z = 1, and R1 = 0.0345. [Co3(dca)6(H2O)4].2(2,3,5-tmpdo) (2) is in the monoclinic space group C2/c with a = 29.477(1), b = 7.3735(2), c = 17.4631(7) A, beta = 93.652(1) degrees , V = 3787.9(2) A(3), Z = 4, and R1 = 0.0532. [Co(dca)2(H2O)2].2(2,3,5,6-tmpdo) (3) is in the monoclinic space group P2(1)/c with a = 9.4739(3), b = 11.3876(3), c = 12.1778(3) A, beta = 98.967(1) degrees , V = 1297.74(6) A(3), Z = 4, and R1 = 0.0481. 1 contains an unusual (4,4) Co-dca layer, representing a rare example of metal-dca coordination polymers with mixing 1,3-mu(2)- and 1,5-mu2-dca bridges. The Co-dca (4, 4) layers are connected by 2,5-dmpdo to give an alpha-Po-type network, which displays antiferromagnetic ordering below 10.8 K. 2 is a unique 3D framework composed of 2D twinned 1,5-mu2-dca bridged layers, which are connected by Co-H2O-Co linkages. The noncoordinated 2,3,5-tmpdo units and the coordination water molecules form hydrogen-bonded chains that thread the framework. 3 has alternating stacks of usual Co-dca (4, 4) layers through 1,5-mu2-dca bridges and organic layers of noncoordinated 2,3,5,6-tmpdo. Weak antiferromagnetic and weak ferromagnetic coupling was observed in 2 and 3, respectively. PMID- 15792452 TI - Tb4FeGe8 grown in liquid gallium: trans-cis chains from the distortion of a planar Ge square net. AB - The ternary germanide Tb4FeGe8 was obtained from Ga flux reactions. The crystal structure studied with single-crystal X-ray diffraction revealed the existence of an orthorhombic average substructure (Cmcm, Z=1) with cell parameters a = 4.1118(14) A, b=15.844(5) A, and c=3.9885(15) A. The refinement [I > 2sigma(I)] converged to final residuals R1/wR2 = 0.0363/0.0893. The average structure (CeNiSi2-type) consists of a 3D [Fe1/4Ge2] framework where Ge atoms form a square net and Fe atoms reside alternatively above and below it with only 1/4 occupation probability. X-ray and electron diffraction studies showed a modulation in the Ge net. The modulated structure was refined based on a 4-fold monoclinic supercell (P2(1)/n) with parameters a = 5.7315(11) A, b = 15.842(3) A, c = 11.438(2) A, and beta = 91.724(4) degrees with R1/wR2 = 0.0643/0.1735 and uncovered a severe distortion of the Ge square net. The Ge atoms are displaced to form an array of cis-trans chains. The Ge-Ge distances within these chains are distinctively bonding, whereas those between the chains are nonbonding. Results of the electronic structure calculations and magnetic measurements are also reported. The structural distortions found in Tb4FeGe8 cast a doubt onto the correctness of many of the reported REM1-xGe2 disordered compounds and call for reinvestigation. PMID- 15792453 TI - Complex magnetic ordering in Eu3InP3: a new rare earth metal zintl compound. AB - Eu3InP3 has been prepared as large single crystals with an indium flux reaction. The structure of the new compound is isotypic to Sr3InP3 and crystallizes in the orthorhombic space group Pnma with unit cell dimensions of a = 12.6517(15) A, b = 4.2683(5) A, and c = 13.5643(14) A (Z = 4, T = 140 K, R1 = 0.0404, wR2 = 0.0971 for all data). The structure consists of one-dimensional chains of corner-shared distorted [InP2P2/2]6- tetrahedra separated by rows of Eu2+ ions. Two of the three crystallographically distinct europium sites have a short Eu(1)-Eu(2) distance of 3.5954(7) A, which yields Eu-Eu dimers. The Eu-P bond distances range from 2.974(2) to 3.166(2) A. The temperature dependence of the conductivity indicates that Eu3InP3 is a small band gap semiconductor. Both magnetization and Eu-151 Mossbauer spectral measurements indicate that the europium in Eu3InP3 is divalent and that at least two magnetic transitions occur. Magnetization studies reveal magnetic transitions at 14, 10.4, and approximately 5 K. These transitions are also observed in heat capacity studies of Eu3InP3. The Mossbauer spectra indicate that the two europium sites are ordered at 12 K and that all three europium sites are ordered at 8 K. PMID- 15792454 TI - Rhenium tricarbonyl core complexes of thymidine and uridine derivatives. AB - Thymidine and uridine were modified at the C2' and C5' ribose positions to form amine analogues of the nucleosides (1 and 4). Direct amination with NaBH(OAc)3 in DCE with the appropriate aldehydes yielded 1-{5-[(bis(pyridin-2 ylmethyl)amino)methyl]-4-hydroxytetrahydrofuran-2-yl}-5-methyl-1H-pyrimidine-2,4 dione (L1), 1-{5-[(bis(quinolin-2-ylmethyl)amino)methyl]-4-hydroxytetrahydrofuran 2-yl}-5-methyl-1H-pyrimidine-2,4-dione (L2), and 1-[3-(bis(pyridin-2 ylmethyl)amino)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-1H-pyrimidine 2,4-dione (L5), while standard coupling procedures of 1 and 4 with 5-(bis(pyridin 2-ylmethyl)amino)pentanoic acid (2) and 5-(bis(quinolin-2 ylmethyl)amino)pentanoic acid (3) in the presence of HOBT-EDCI in DMF provided a second novel series of bifunctional chelators: 5-(bis(pyridin-2 ylmethyl)amino)pentanoic acid [(3-hydroxy-5-(5-methyl-4-oxo-3,4-dihydro-2H pyrimidin-1-yl)tetrahydrofuran-2-yl)methyl] amide (L3), 5-(bis(quinolin-2 ylmethyl)amino)pentanoic acid [(3-hydroxy-5-(5-methyl-4-oxo-3,4-dihydro-2H pyrimidin-1-yl)tetrahydrofuran-2-yl)methyl] amide (L4), 5-(bis(pyridin-2 ylmethyl)amino)pentanoic acid [2-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-4 hydroxy-5-(hydroxymethyl)tetrahydrofuran-3-yl] amide (L6), and 5-(bis(quinolin-2 ylmethyl)amino)pentanoic acid [2-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-4 hydroxy-5-(hydroxymethyl)tetrahydrofuran-3-yl] amide (L7). The rhenium tricarbonyl complexes of L1-L4, L6, and L7, [Re(CO)3(LX)]Br (X=1-4, 6, 7: compounds 5-10, respectively), have been prepared by reacting the appropriate ligand with [NEt4][Re(CO)3Br3] in methanol. The ligands and their rhenium complexes were obtained in good yields and characterized by common spectroscopic techniques including 1D and 2D NMR, HRMS, IR, cyclic voltammetry, UV, and luminescence spectroscopy and X-ray crystallography. The crystal structure of complex 6.0.5NaPF6 displays a facial geometry of the carbonyl ligands. The nitrogen donors of the tridentate ligand complete the distorted octahedral spheres of the complex. Crystal data: monoclinic, C2, a = 24.618(3) A, b = 11.4787(11) A, c = 15.5902(15) A, beta = 112.422(4) degrees , Z = 4, D(calc) = 1.562 g/cm3. PMID- 15792455 TI - Structural trends and chemical bonding in Te-doped silicon clathrates. AB - The recently discovered tellurium-doped silicon clathrates, Te7+xSi20-x and Te16Si38, both low- and high-temperature forms (cubic and rhombohedral, respectively), exhibit original structures that are all derived from the parent type I clathrate G8Si46 (G = guest atom). The similarities and differences between the structures of these compounds and that of the parent one are analyzed and discussed on the basis of charge distribution and chemical bonding considerations. Because of the particular character of the Te atom, these compounds appear to be at the border between the clathrate and polytelluride families. PMID- 15792456 TI - Variation of oxo-transfer reactivity of (nitro)cobalt picket fence porphyrin with oxygen-donating ligands. AB - Derivatives of (nitro)cobalt picket fence porphyrin with oxygen-donating ligands have been prepared in solution and in the solid state. Crystal structures of two of these derivatives, (H2O)CoTpivPP(NO2) and (CH3OH)CoTpivPP(NO2), have been determined. The ethanol complex (C2H5OH)Co(TPP)(NO2) has been obtained and spectrally characterized using sublimed layers methodology. The formation constant and the DeltaH degrees value of the association reaction with ethanol have been determined by FTIR measurements in CCl4 solution. Catalytic oxygen activation and oxo-transfer reactions of these derivatives have been assessed in solution. Correlations between the oxo-transfer reactivity, thermodynamics, and characteristics of the nitro ligand show that although calculated and observed ONO vibrational spectra and bond lengths suggest activation of the NO2 ligand and enhanced oxo-transfer reactions as seen in the analogous five-coordinate complexes, density functional theory calculations support that thermodynamics limits oxo-atom transfer reactions in these six-coordinate systems. PMID- 15792457 TI - Lead(II) thiocyanate complexes with bibracchial lariat ethers: an X-ray and DFT study. AB - Compounds of formula [Pb(L2)(NCS)2] (1) and [Pb(L4)(SCN)2] (2) (where L2 is the lariat crown ether N,N'-bis(3-aminobenzyl)-4,13-diaza-18-crown-6 and L4 is the Schiff-base lariat crown ether N,N'-bis(3-(salicylaldimino)benzyl)-4,13-diaza-18 crown-6) were isolated and structurally characterized by X-ray diffraction analyses. The X-ray crystal structures of both compounds show the metal ion coordinated to the six donor atoms of the crown moiety, leaving the corresponding pendant arms uncoordinated. The coordination sphere of lead(II) is completed by two thiocyanate groups that coordinate either through their nitrogen (1) or sulfur (2) atoms. The organic receptor adopts a syn conformation in 1, while in 2 it shows an anti conformation. To rationalize these unexpected different conformations of the L2 and L4 receptors in compounds 1 and 2, as well as the different binding modes found for the thiocyanate ligands, we have carried out theoretical calculations at the DFT (B3LYP) level. These calculations predict the syn conformation being the most stable in both 1 and 2 complexes. So, the anti conformation found for 2 in the solid state is tentatively attributed to the presence of intermolecular pi-pi interactions between phenol rings, for which the dihedral angle between the least-squares planes of both rings amounts to 2.6 degrees and the distance between the center of both rings is 3.766 A. On the other hand, the analysis of the electronic structure has revealed that the Pb ligand bonds present highly ionic character in this family of compounds. They also suggest a greater transfer of electron density from the NCS- ligands when they coordinate through the less electronegative S atom. The Pb-SCN covalent bond formation mainly occurs due to an effective overlap of the occupied 3p z orbitals of the S atoms and the unoccupied 6p z AO of the Pb atom, while the Pb-NCS bonding interaction is primarily due to the overlap of the 6s and 7s AO of Pb with sp(1.10) hybrids of the N donor atoms. Our electronic structure calculations can rationalize the different coordination of the thiocyanate groups in compounds 1 and 2: the simultaneous formation of two Pb-SCN bonds is more favorable for S Pb-S angles close to 180 degrees , for which the overlap between the occupied 3p z orbitals of the S atoms and the unoccupied 6 pz AO of the Pb atom is maximized. PMID- 15792459 TI - Performance of nonrelativistic and quasi-relativistic hybrid DFT for the prediction of electric and magnetic hyperfine parameters in 57Fe Mossbauer spectra. AB - 57Fe electric and magnetic hyperfine parameters were calculated for a series of 10 iron model complexes, covering a wide range of oxidation and spin states. Employing the B3LYP hybrid method, results from nonrelativistic density functional theory (DFT) and quasi-relativistic DFT within the zero-order regular approximation (ZORA) were compared. Electron densities at the iron nuclei were calculated and correlated with experimental isomer shifts. It was shown that the fit parameters do not depend on a specific training set of iron complexes and are, therefore, more universal than might be expected. The nonrelativistic and quasi-relativistic electron densities gave fit parameters of similar quality; the ZORA densities are only shifted by a factor of 1.32, upward in the direction of the four-component Dirac-Fock value. From a correlation of calculated electric field gradients and experimental quadrupole splittings, the value of the 57Fe nuclear quadrupole moment was redetermined to a value of 0.16 barn, in good agreement with other studies. The ZORA approach gave no additional improvement of the calculated quadrupole splittings in comparison to the nonrelativistic approach. The comparison of the calculated and measured 57Fe isotropic hyperfine coupling constants (hfcc's) revealed that both the ZORA approach and the inclusion of spin-orbit contributions lead to better agreement between theory and experiment in comparison to the nonrelativistic results. For all iron complexes with small spin-orbit contributions (high-spin ferric and ferryl systems), a distinct underestimation of the isotropic hfcc's was found. Scaling factors of 1.81 (nonrelativistic DFT) and 1.69 (ZORA) are suggested. The calculated 57Fe isotropic hfcc's of the remaining model systems (low-spin ferric and high-spin ferrous systems) contain 10-50% second-order contributions and were found to be in reasonable agreement with the experimental results. This is assumed to be the consequence of error cancellation because g-tensor calculations for these systems are of poor quality with the existing DFT approaches. Excellent agreement between theory and experiment was found for the 57Fe anisotropic hfcc's. Finally, all of the obtained fit parameters were used for an application study of the [Fe(H2O)6]3+ ion. The calculated spectroscopic data are in good agreement with the Mossbauer and electron paramagnetic resonance results discussed in detail in a forthcoming paper. PMID- 15792458 TI - Solid-supported synthesis of polymerizable lanthanide-ion chelating lipids for protein detection. AB - Usually, lipids are synthesized employing solution-phase organic synthesis techniques. Though successful, the purifications can be difficult to accomplish due to the amphiphilic nature of the molecules. Herein, we demonstrate the advantages of a solid-phase approach for preparing a variety of metal-chelating lipids. A number of saturated and polymerizable metal-chelating lipids were prepared using this methodology. This approach requires one chromatographic purification after cleaving the lipids from the solid support. We also demonstrate that the resulting polymerized liposomes (containing Eu3+) possess the appropriate luminescence properties for the qualitative and quantitative determination of proteins. PMID- 15792460 TI - Theoretical investigations of uranyl-ligand bonding: four- and five-coordinate uranyl cyanide, isocyanide, carbonyl, and hydroxide complexes. AB - The coordination and bonding of equatorial hydroxide, carbonyl, cyanide (CN-), and isocyanide (NC-) ligands with uranyl dication, [UO2]2+, has been studied using density functional theory with relativistic effective core potentials. Good agreement is seen between experimental and calculated geometries of [UO2(OH)4]2-. Newly predicted ground-state structures of [UO2(OH)5]3-, [UO2(CO)4]2+, [UO2(CO)5]2+, [UO2(CN)4]2-, [UO2(CN)5]3-, [UO2(NC)4]2-, and [UO2(NC)5]3- are reported. Four-coordinate uranyl isocyanide complexes are the predicted gas-phase species while five-coordinate uranyl cyanide complexes are energetically favorable in aqueous solution. Small energy differences between cyanide and isocyanide complexes indicate the energetic feasibility of mixed cyanide and isocyanide complexes. A D2d uranyl tetrahydroxide is the dominant gas-phase and aqueous species, but formation of uranyl carbonyl complexes is seen to be exothermic in the gas-phase and endothermic in aqueous solution. PMID- 15792461 TI - New d0 transition metal iodates: synthesis, structure, and characterization of BaTi(IO3)6, LaTiO(IO3)5, Ba2VO2(IO3)4.(IO3), K2MoO2(IO3)4, and BaMoO2(IO3)4.H2O. AB - Five new d0 transition metal iodates, BaTi(IO3)6, LaTiO(IO3)5, Ba2VO2(IO3)4.(IO3), K2MoO2(IO3)4, and BaMoO2(IO3)4.H2O, have been synthesized by hydrothermal methods using Ba(OH)2.8H2O, La2O3, K2CO3, TiO2, V2O5, MoO3, and HIO3 as reagents. The structures of these compounds were determined by single-crystal X-ray diffraction. All of the reported materials have zero-dimensional or pseudo one-dimensional crystal structures composed of MO6 (M = Ti4+, V5+, or Mo6+) octahedra connected to IO3 polyhedra. Infrared and Raman spectroscopy, thermogravimetric analysis, and UV-vis diffuse reflectance spectroscopy are also presented. Crystal data: BaTi(IO3)6, trigonal, space group R-3 (No. 148), with a = b = 11.4711(10) A, c = 11.1465(17) A, V = 1270.2(2) A3, and Z = 3; LaTiO(IO3)5, monoclinic, space group P2(1)/n (No. 14), with a = 7.4798(10) A, b = 18.065(2) A, c = 10.4843(14) A, beta = 91.742(2) degrees , V = 1416.0(3) A3, and Z = 4; Ba2VO2(IO3)4.(IO3), monoclinic, space group P2(1)/c (No. 14), with a = 7.5012(9) A, b = 33.032(4) A, c = 7.2150(9) A, beta = 116.612(2) degrees , V = 1598.3(3) A3, and Z = 4; K2MoO2(IO3)4, monoclinic, space group C2/c (No. 15), with a = 12.959(2) A, b = 6.0793(9) A, c = 17.748(3) A, beta = 102.410(4) degrees , V = 1365.5(4) A3, and Z = 4; BaMoO2(IO3)4.H(2)O, monoclinic, space group P2(1)/n (No. 14), with a = 13.3368(17) A, b = 5.6846(7) A, c = 18.405(2) A, beta = 103.636(2) degrees , V = 1356.0(3) A3, and Z = 4. PMID- 15792462 TI - Structure and physical properties of ternary rare-earth cobalt bismuth intermetallics (RE)12Co5Bi (RE = Y, Gd, Tb, Dy, Ho, Er, Tm). AB - The rare-earth intermetallic compounds (RE)12Co5Bi (RE = Y, Gd, Tb, Dy, Ho, Er, Tm) were prepared by arc-melting and annealing at 600 degrees C. These compounds extend the previously known (RE)6M2+xX1-x (M = Co, Ni; X = Ga, In, Sn, Pb) series with the Ho6Co2Ga-type structure to X = Bi. The crystal structure of Ho12Co5Bi was refined by the Rietveld method from powder X-ray diffraction data obtained using synchrotron radiation (Pearson symbol oI36, orthorhombic, space group Immm, Z = 2, a = 9.37598(14) A, b = 9.37871(14) A, c = 9.85465(13) A). Unlike other Ho6Co2Ga-type compounds, the 2a site in Ho12Co5Bi is exclusively occupied by Co atoms. Four-probe electrical resistivity measurements on sintered polycrystalline samples of (RE)12Co5Bi indicated metallic behavior. Magnetic measurements revealed behavior ranging from frequency-dependent maxima in the ac susceptibility for Y12Co5Bi to possible ferrimagnetic ordering for Gd12Co5Bi to antiferromagnetic ordering with metamagnetic transitions for the remaining compounds. As confirmed by band structure calculations using Y12Co5Bi as a model compound, Y-Y and Y-Co interactions are the most important bonding components, but matrix effects are likely responsible for anomalously short Co-Co contacts in the structure. PMID- 15792463 TI - Structure and properties of the thorium vanadyl tellurate, Th(VO2)2(TeO6)(H2O)2. AB - The hydrothermal reaction of Th(NO3)4.xH2O with V2O5 and H6TeO6 at 200 degrees C under autogenously generated pressure results in the formation of Th(VO2)2(TeO6)(H2O)2 as a pure phase. The single-crystal X-ray data indicate that Th(VO2)2(TeO6)(H2O)2 possesses a three-dimensional structure constructed from ThO9 tricapped trigonal prisms, VO5 distorted square pyramids, VO4 distorted tetrahedra, and TeO6 distorted octahedra. Both of the vanadium polyhedra contain VO2+ vanadyl units with two short V=O bond distances. The tellurate octahedron is tetragonally distorted and utilizes all of its oxygen atoms to bond to adjacent metal centers, sharing edges with ThO9 and VO5 units, and corners with two ThO9, one VO5, and two VO4 polyhedra. Crystallographic data: Th(VO2)2(TeO6)(H2O)2, orthorhombic, space group Pbca, a = 12.6921(7), b = 11.5593(7), c = 13.0950(8) A, Z = 8 (T = 193 K). The UV-vis diffuse reflectance spectrum of Th(VO2)2(TeO6)(H2O)2 shows vanadyl-based charge-transfer absorption features. Th(VO2)2(TeO6)(H2O)2 decomposes primarily to Th(VO3)4 when heated at 600 degrees C in air. PMID- 15792464 TI - Synthesis and characterization of hydrogen-bonded assemblies of W6S8L6 clusters. AB - Ligand-exchange reactions involving octahedral W6S8 clusters and a family of pyridine-based ligands (isonicotinic acid, isonicotinamide, 4-hydroxypyridine, 4 aminopyridine, 4-pyridineacetamide) have been explored with the goal of preparing compounds that crystallize in hydrogen-bonded arrays. Two new compounds, W6S8(4 pyridineacetamide)6.DMF.4-pyridineacetamide (1) and W6S8(4-aminopyridine)6.4DMF (2), were isolated and characterized by single-crystal X-ray diffraction. Both compounds crystallize in the P2(1)/c space group with a = 16.461(1), b = 33.08(2), c = 13.165(10) A, beta = 103.270(15) degrees for 1 and a = 13.8988(5), b = 13.2791(5), c = 15.6293(6) A, beta = 108.5410(10) degrees for 2. Each compound was further characterized by 1H NMR spectroscopy, elemental (CHN) analysis, and thermogravimetric analysis. Examination of the structures shows that 1 forms a three-dimensional hydrogen-bonded network in which each 4 pyridineacetamide ligand interacts with ligands on neighboring clusters or with the free ligand of crystallization. This is the first hydrogen-bonded network formed from W6S8 clusters. In 2, the amino groups act as hydrogen-bond donors toward DMF molecules of crystallization, but an extended array is not formed. In addition, the binding strengths of these pyridine-based ligands to the W6S8 cluster were studied through quantitative 1H NMR studies of ligand-exchange reactions. A qualitative relationship was found between ligand binding strengths and Hammett substituent constants for this group of ligands. PMID- 15792465 TI - Photophysical, spectroscopic, and computational studies of a series of Re(I) tricarbonyl complexes containing 2,6-dimethylphenylisocyanide and 5- and 6 derivatized phenanthroline ligands. AB - The ligand 2,6-dimethylphenylisocyanide (CNx) forms six complexes of the formula [Re(CO)3(CNx)(L)]+, where L = 1,10-phenanthroline (1), 5-chloro-1,10 phenanthroline (2), 5-nitro-1,10-phenanthroline (3), 5-methyl-1,10-phenanthroline (4), 5,6-dimethyl-1,10-phenanthroline (5), and 1,10-phenanthrolinopyrrole (6). The lowest-energy absorption peaks of the complexes red-shift in the order 1 < 2 < 3 < 4 < 5 < 6. The time-dependent density functional theory (TDDFT) and conductor-like polarizable continuum model (CPCM) computed singlet excited states in ethanol deviate by 1000 cm(-1) or less from the experimental UV-vis peaks. The complexes undergo reversible reductions and irreversible oxidations. The electronic energy gap increases in the order 3 < 2 < 1 < 4 < 5 < 6, which is the order of increasing electron-donating power of the phen substituents. The reduction potentials linearly correlate with the B3LYP calculated LUMO energies for 1-6. The complexes emit at room temperature and at 77 K except 3, which emits only at 77 K. The calculated (3)MLLCT energies are within 1100 cm(-1) from the experimental emission energies at 77 K. The 77 K emission curve-fitting analysis results agree with the computational assignment of the emitting state as 3MLLCT for 1-5 and 3LC for 6. The experimental 77 K emission energies and the calculated 3MLLCT state energies increase in the order 6 < 5, 3 < 2 < 4, 1. The 77 K emission lifetimes increase upon addition of substituents from 65 micros for 1 to 171 micros for 2, to 230 micros for 4 and 5, and to 322 micros for 3. The emission quantum yields at room temperature in solution are 0.77, 0.78, 0.83, 0.56, and 0.11 for complexes 1, 2, 4, 5, and 6, respectively. PMID- 15792467 TI - Behavior of tri(n-butyl)ammonium bis[citrato(3-)-O1,O3,O6]silicate in aqueous solution: analysis of a sol-gel process by small-angle neutron scattering. AB - The racemic hexacoordinate silicon(IV) complex tri(n-butyl)ammonium bis[citrato(3 )-O1,O3,O6]silicate (1) was synthesized by treatment of Si(OMe)4 with 2 molar equiv of citric acid and 2 molar equiv of N(n-Bu)3. The corresponding germanium analogue, tri(n-butyl)ammonium bis[citrato(3-)-O1,O3,O6]germanate (5; structurally characterized by single-crystal X-ray diffraction), was obtained analogously, starting from Ge(OMe)4. Upon dissolution in water, the lambda6Si silicate dianion of 1 hydrolyzes spontaneously (formation of Si(OH)4 and citric acid), whereas the lambda6Ge-germanate dianion of 5 was found to be stable in water. Aqueous "solutions" of 1, with concentrations that are significantly higher than the saturation concentration of Si(OH)4, look absolutely clear over a period of several weeks; however, in reality, these solutions are sols with very small particles that slowly grow with time and finally form a gel that precipitates. This sol-gel process was monitored by small-angle neutron scattering (SANS). For reasons of comparison, an aqueous solution of the hydrolytically stable germanium compound 5 was also studied by the SANS technique. PMID- 15792466 TI - Efficient plasmid DNA cleavage by a mononuclear copper(II) complex. AB - The Cu(II) complex of the ligand all-cis-2,4,6-triamino-1,3,5 trihydroxycyclohexane (TACI) is a very efficient catalyst of the cleavage of plasmid DNA in the absence of any added cofactor. The maximum rate of degradation of the supercoiled plasmid DNA form, obtained at pH 8.1 and 37 degrees C, in the presence of 48 microM TACI.Cu(II), is 2.3 x 10(-3) s(-1), corresponding to a half life time of only 5 min for the cleavage of form I (supercoiled) to form II (relaxed circular). The dependence of the rate of plasmid DNA cleavage from the TACI.Cu(II) complex concentration follows an unusual and very narrow bell-like profile, which suggests an high DNA affinity of the complexes but also a great tendency to form unreactive dimers. The reactivity of the TACI.Cu(II) complexes is not affected by the presence of several scavengers for reactive oxygen species or when measured under anaerobic conditions. Moreover, no degradation of the radical reporter Rhodamine B is observed in the presence of such complexes. These results are consistent with the operation of a prevailing hydrolytic pathway under the normal conditions used, although the failure to obtain enzymatic religation of the linearized DNA does not allow one to rule out the occurrence of a nonhydrolytic oxygen-independent cleavage. A concurrent oxidative mechanism becomes competitive upon addition of reductants or in the presence of high levels of molecular oxygen: under such conditions, in fact, a remarkable increase in the rate of DNA cleavage is observed. PMID- 15792468 TI - Architecture of supramolecular metal complexes for photocatalytic CO2 reduction: ruthenium-rhenium bi- and tetranuclear complexes. AB - We study the electrochemical, spectroscopic, and photocatalytic properties of a series of Ru(II)-Re(I) binuclear complexes linked by bridging ligands 1,3-bis(4' methyl-[2,2']bipyridinyl-4-yl)propan-2-ol (bpyC3bpy) and 4-methyl-4' [1,10]phenanthroline-[5,6-d]imidazol-2-yl)bipyridine (mfibpy) and a tetranuclear complex in which three [Re(CO)3Cl] moieties are coordinated to the central Ru using the bpyC3bpy ligands. In the bpyC3bpy binuclear complexes, 4,4'-dimethyl 2,2'-bipyridine (dmb) and 4,4'-bis(trifluoromethyl)-2,2'-bipyridine ({CF3}2bpy), as well as 2,2'-bipyridine (bpy), were used as peripheral ligands on the Ru moiety. Greatly improved photocatalytic activities were obtained only in the cases of [Ru{bpyC3bpyRe(CO)3Cl}3]2+ (RuRe3) and the binuclear complex [(dmb)2Ru(bpyC3bpy)Re(CO)3Cl]2+ (d2Ru-Re), while photocatalytic responses were extended further into the visible region. The excited state of ruthenium in all Ru-Re complexes was efficiently quenched by 1-benzyl-1,4-dihydronicotinamide (BNAH). Following reductive quenching in the case of d2Ru-Re, generation of the one-electron-reduced (OER) species, for which the added electron resides on the Ru-bound bpy end of the bridging ligand bpyC3bpy, was confirmed by transient absorption spectroscopy. The reduced Re moiety was produced via a relatively slow intramolecular electron transfer, from the reduced Ru-bound bpy to the Re site, occurring at an exchange rate (DeltaG approximately 0). Electron transfer need not be rapid, since the rate-determining process is reduction of CO2 with the OER species of the Re site. Comparison of these results with those for other bimetallic systems gives us more general architectural pointers for constructing supramolecular photocatalysts for CO2 reduction. PMID- 15792469 TI - Synthesis and structural characterization of novel neutral hexacoordinate silicon(IV) complexes with SiO2N4 skeletons containing cyanato-N or thiocyanato-N ligands. AB - A series of novel hexacoordinate silicon(IV) complexes with an SiO2N4 skeleton (compounds (OC-6-12)-3, (OC-6-12)-4, (OC-6-12)-5, (OC-6-12)-6, and (OC-6-2'2)-7) were synthesized, starting from Si(NCO)4 or Si(NCS)4. These compounds contain (i) two bidentate O,N-chelate ligands (or one tetradentate O,N,N,O-chelate ligand) derived from 4-aminopent-3-en-2-ones of the formula type Me-C(NRH)=CH-C(O)-Me (R = organyl) and (ii) two monodentate cyanato-N or thiocyanato-N ligands. Formally, the bidentate singly negatively charged O,N-chelate ligands (tetradentate 2-fold negatively charged O,N,N,O-chelate ligand) behave as ligands of the imino-enolato type. In addition, the adduct trans-8 was synthesized by reaction of Si(NCS)4 with 2 molar equiv of Me-C(Ni-PrH)=CH-C(O)-Me. This hexacoordinate silicon(IV) complex contains (i) four monodentate thiocyanato-N ligands and (ii) two neutral monodentate ligands of the iminio-enolato type. All compounds synthesized were structurally characterized by single-crystal X-ray diffraction and solid-state and solution NMR spectroscopy. To get more information about the stereochemistry of these compounds, the experimental investigations were complemented by computational studies. PMID- 15792470 TI - Solubility, complex formation, and redox reactions in the Tl2O3-HCN/CN(-)-H2O system. Crystal structures of the cyano compounds Tl(CN)3.H2O, Na[Tl(CN)4].3H2O, K[Tl(CN)4], and Tl(I)[Tl(III)(CN)4] and of Tl(I)2C2O4. AB - Thallium(III) oxide can be dissolved in water in the presence of strongly complexing cyanide ions. Tl(III) is leached from its oxide both by aqueous solutions of hydrogen cyanide and by alkali-metal cyanides. The dominating cyano complex of thallium(III) obtained by dissolution of Tl2O3 in HCN is [Tl(CN)3(aq)] as shown by 205Tl NMR. The Tl(CN)3 species has been selectively extracted into diethyl ether from aqueous solution with the ratio CN-/Tl(III) = 3. When aqueous solutions of the MCN (M = Na+, K+) salts are used to dissolve thallium(III) oxide, the equilibrium in liquid phase is fully shifted to the [Tl(CN)4]- complex. The Tl(CN)3 and Tl(CN)4- species have for the first time been synthesized in the solid state as Tl(CN)3.H2O (1), M[Tl(CN)4] (M = Tl (2) and K (3)), and Na[Tl(CN)4].3H2O (4) salts, and their structures have been determined by single-crystal X-ray diffraction. In the crystal structure of 1, the thallium(III) ion has a trigonal bipyramidal coordination with three cyanide ions in the equatorial plane, while an oxygen atom of the water molecule and a nitrogen atom from a cyanide ligand, attached to a neighboring thallium complex, form a linear O-Tl-N fragment. In the three compounds of the tetracyano thallium(III) complex, 2-4, the [Tl(CN)4]- unit has a distorted tetrahedral geometry. Along with the acidic leaching (enhanced by Tl(III)-CN- complex formation), an effective reductive dissolution of the thallium(III) oxide can also take place in the Tl2O3-HCN-H2O system yielding thallium(I), while hydrogen cyanide is oxidized to cyanogen. The latter is hydrolyzed in aqueous solution giving rise to a number of products including (CONH2)2, NCO-, and NH4+ detected by 14N NMR. The crystalline compounds, Tl(I)[Tl(III)(CN)4], Tl(I)2C2O4, and (CONH2)2, have been obtained as products of the redox reactions in the system. PMID- 15792471 TI - Structural, magnetic, and electrical characterization of new polycrystalline phases of nickel- and platinum-doped [(DT-TTF)n][Au(mnt)2] (n = 1, 2). AB - Doping of spin-ladder systems by isostructural paramagnetic complexes was attempted. Despite the close isostructural nature of the pure (DT-TTF)2[M(mnt)2] (M = Au, Ni, Pt) end-members, which present a ladder structure, doping of the spin-ladder (DT-TTF)2[Au(mnt)2] with either 5% or 25% [M(mnt)2]- (M = Ni, Pt) generates two (metrically) new phases. Their markedly different crystal structures have been determined using laboratory X-ray powder diffraction data. (DT-TTF)2[Au0.75Ni0.25(mnt)2] consists of a mixed-valence compound (of triclinic symmetry), which was only detected, pure or in a mixture of phases, when [Ni(mnt)2]- was used as a dopant. Differently, the stoichiometric 1:1 [DT TTF][Au0.75Pt0.25(mnt)2] monoclinic phase was found when [Pt(mnt)2]- (in 5% and 25%) was employed as the doping agent. Remarkably, only in the 5% Pt doping experiment, the major component of the mixture was the ladder structure compound (DT-TTF)2[Au(mnt)2] doped with minor amounts of Pt. This 5% Pt-doped specimen shows an EPR signal (g = 2.0115, DeltaHpp = 114 G at 300 K) wider than the pure compound (DT-TTF)2[Au(mnt)2], denoting exchange between the donor spins and Pt(mnt)2- centers. The electrical transport properties of the 5% Pt-doped composition at high temperatures are comparable to those of (DT-TTF)2[Au(mnt)2] with room-temperature conductivity sigma300K = 13 S/cm and thermopower S300K = 46 microV/K, with a sharp transition at 223 K similar to that previously observed in the Cu analogue at 235 K. PMID- 15792472 TI - Mechanistic insights from reactions between copper(II)-phenoxyl complexes and substrates with activated C-H bonds. AB - The reactivities of two copper(II)-phenoxyl analogues of the oxidized, active form of the metalloenzyme galactose oxidase, [1tBu2]+ and [2tBu2]+, have been studied using the substrates benzyl alcohol and 9,10-dihydroanthracene, for a total of four reactions. The reaction stoichiometries in all cases show a 2:1 ratio of oxidant to benzaldehyde or anthracene product, indicating that [1tBu2]+ and [2tBu2]+ behave ultimately as only one-electron oxidants, but the reaction kinetics each indicate that only a single copper(II)-phenoxyl complex is involved in the rate-determining step. For each substrate, rate laws indicate that [1tBu2]+ and [2tBu2]+ react by different mechanisms: one proceeds by a simple bimolecular reaction, while the other first enters into a substrate-binding equilibrium before subsequently reacting by an intramolecular reaction. The reactions proceeding by the latter mechanism have faster overall rates, which correlates to a lower entropic barrier for the substrate-binding mechanism. Correlation of the reaction rates with the C-H bond dissociation energies of substrates as well as significant deuterium kinetic isotope effects indicates that the rate-determining steps involve hydrogen atom abstraction from the activated C-H bonds. A variable-temperature study (268-308 K) of the nonclassical KIE of the [1tBu2]+/benzyl alcohol reaction (kH/kD = 15 at 298 K) failed to show evidence for quantum tunneling. The rapid sequence by which a second 1 equiv of copper(II)-phenoxyl oxidant completes the reaction after the rate- and product determining hydrogen atom abstraction step cannot be probed kinetically. Comparisons are made to the reactivities of other copper(II)-phenoxyl complexes reported in the literature and to galactose oxidase itself. PMID- 15792473 TI - Copper hydroxydiphosphate with a one-dimensional arrangement of copper polyhedra: Cu3[P2O6OH]2. AB - A new copper hydroxydiphosphate Cu3(P2O6OH)2 was synthesized, by soft chemistry. The crystal structure was solved ab initio from X-ray powder diffraction data in the triclinic space group P. The structure is built up from [Cu3O10]infinity zigzag chains linked by P2O6(OH) groups to form a tridimensional framework. The [Cu3O10]infinity chains consist of edge-sharing polyhedra. The structure contains two sorts of copper polyhedra: one CuO6 octahedron and two CuO5 pyramids. Magnetization measurements confirm the presence of divalent copper and suggest antiferromagnetic interactions at low temperature. PMID- 15792474 TI - Reactivity of rhenium(V) oxo Schiff base complexes with phosphine ligands: rearrangement and reduction reactions. AB - The symmetric rhenium(V) oxo Schiff base complexes trans-[ReO(OH2)(acac2en)]Cl and trans-[ReOCl(acac2pn)], where acac2en and acac2pn are the tetradentate Schiff base ligands N,N'-ethylenebis(acetylacetone) diimine and N,N' propylenebis(acetylacetone) diimine, respectively, were reacted with monodentate phosphine ligands to yield one of two unique cationic phosphine complexes depending on the ligand backbone length (en vs pn) and the identity of the phosphine ligand. Reduction of the Re(V) oxo core to Re(III) resulted on reaction of trans-[ReO(OH2)(acac2en)]Cl with triphenylphosphine or diethylphenylphosphine to yield a single reduced, disubstituted product of the general type trans [Re(III)(PR3)2(acac2en)]+. Rather unexpectedly, a similar reaction with the stronger reducing agent triethylphosphine yielded the intramolecularly rearranged, asymmetric cis-[Re(V)O(PEt3)(acac2en)]+ complex. Reactions of trans [Re(V)O(acac2pn)Cl] with the same phosphine ligands yielded only the rearranged asymmetric cis-[Re(V)O(PR3)(acac2pn)]+ complexes in quantitative yield. The compounds were characterized using standard spectroscopic methods, elemental analyses, cyclic voltammetry, and single-crystal X-ray diffraction. The crystallographic data for the structures reported are as follows: trans [Re(III)(PPh3)2(acac2en)]PF6 (H48C48N2O2P2Re.PF6), 1, triclinic (P), a = 18.8261(12) A, b = 16.2517(10) A, c = 15.4556(10) A, alpha = 95.522(1) degrees , beta = 97.130(1) degrees , gamma = 91.350(1) degrees , V = 4667.4(5) A(3), Z = 4; trans-[Re(III)(PEt2Ph)2(acac2en)]PF6 (H48C32N2O2P2Re.PF6), 2, orthorhombic (Pccn), a = 10.4753(6) A, b =18.4315(10) A, c = 18.9245(11) A, V = 3653.9(4) A3, Z = 4; cis-[Re(V)O(PEt3)(acac2en)]PF6 (H33C18N2O3PRe.1.25PF6, 3, monoclinic (C2/c), a = 39.8194(15) A, b = 13.6187(5) A, c = 20.1777(8) A, beta = 107.7730(10) degrees , V = 10419.9(7) A3, Z = 16; cis-[Re(V)O(PPh3)(acac2pn)]PF6 (H35C31N2O3PRe.PF6), 4, triclinic (P), a = 10.3094(10) A, b =12.1196(12) A, c = 14.8146(15) A, alpha = 105.939(2) degrees , beta = 105.383(2) degrees , gamma = 93.525(2) degrees , V = 1698.0(3) A3, Z = 2; cis-[Re(V)O(PEt2Ph)(acac2pn)]PF6 (H35C23N2O3PRe.PF6), 5, monoclinic (P2(1)/n), a = 18.1183(18) A, b = 11.580(1) A, c = 28.519(3) A, beta = 101.861(2) degrees , V = 5855.9(10) A(3), Z = 4. PMID- 15792475 TI - Magneto-structural correlations: synthesis of a family of end-on azido-bridged manganese(II) dinuclear compounds with S = 5 spin ground state. AB - The preparation of a series of multidentate pyridyl-imine ligands, L1-L3, and their reactivity with the Mn(II)/N3- system is described (L1 = [N,N-bis(pyridine 2-yl)benzylidene]ethane-1,2-diamine; L2 = [N,N-bis(pyridine-2 yl)benzylidene]propane-1,3-diamine, and L3 = [N,N-bis(pyridine-2 yl)benzylidene]butane-1,4-diamine). Complexes comprising dinuclear end-on bis(mu azido)-bridged manganese(II) units of formulas [Mn2(L1)2(N3)4][Mn2(L1)2(N3)2(CH3OH)2](ClO4)2 (two cocrystallized dinuclear units, 1.2), [Mn2(L2)2(N3)2](ClO4)2 (3), and [Mn2(L3)2(N3)2](ClO4)2 (4) have been synthesized. The crystal structures of complexes 1-4 as well as their magnetic properties are presented. Each manganese atom of cocrystallized complexes in compound 1.2 is heptacoordinated, displaying Mn-N-Mn angles, theta, of 102.53(12) and 101.70(12) degrees and Mn...Mn distances of 3.5091(7) and 3.4680(7) A. On the other hand, each manganese center in compounds 3 and 4 is located within an octahedral coordination environment, the complexes displaying theta angles of 104.29(11) and 103.60(18) degrees , respectively, and Mn...Mn vectors of 3.5371(7) and 3.5338(10) A, respectively. Magnetic susceptibility studies revealed the presence of intramolecular ferromagnetic superexchange, yielding an S = 5 spin ground state in all complexes. Fitting of the experimental data led to coupling constants, intermolecular exchange values, and g factors (in the J/zJ'/g format) of 0.77 cm(-1)/0.01 cm(-1)/2.20 (1.2), 2.04 cm(-1)/0.01 cm(-1)/1.99 (3), and 1.75 cm(-1)/-0.05 cm(-1)/2.04 (4), respectively (using H = -2JS1S2 as the convention for the Heisenberg spin-Hamiltonian). These results are consistent with predictions from recent DFT calculations performed on end-on bis(mu-N3-) bridged Mn(II) dinuclear complexes. A plot of experimental J vs theta, including data from the only preexisting compound of this kind, reveals a linear relationship, which could be the first evidence of a possible magneto-structural correlation between these two parameters. PMID- 15792476 TI - Structures of bisphosphonate metal complexes: zinc and cadmium complexes of clodronate and its partial ester derivatives. AB - Four new zinc and cadmium bisphosphonates [{NaZn(Cl2CP2O6H)(H2O)5}]n (1), [{Cd2(Cl2CP2O6)(H2O)4}.H2O]n (2), [{Zn(Cl2CP2O6Pri2)(H2O)3}.H2O]n (3), and [{Cd2(Cl2CP2O6Pri2)2(MeOH)2(H2O)2}.H2O]2 (4) have been prepared and their crystal structures determined by single-crystal X-ray diffractometry. Two bisphosphonate ligands were used: clodronate, (dichloromethylene)bis(phosphonate) and its symmetrical P,P'-diisopropyl ester derivative. The structure of the Zn complex 1 is three-dimensional, consisting of one-dimensional Zn-clodronate chains connected to the three-dimensional network by Na+ ions. The structure of Cd complex 2 consists of double layers, and a unique bond was found between the Cd2+ cation and a Cl atom of clodronate. Zn complex 3 consists of one-dimensional chains, but the binding of the bisphosphonate ligands is unique: in 3 the bisphosphonate ligand is only bidentate. Compound 4 is a tetramer, and hydrogen bonds hold the tetramers together, forming a layered structure. PMID- 15792477 TI - Analysis of the spin exchange interactions and the ordered magnetic structures of lithium transition metal phosphates LiMPO4 (M = Mn, Fe, Co, Ni) with the olivine structure. AB - The olivine-type compounds LiMPO4 (M = Mn, Fe, Co, Ni) consist of MO4 layers made up of corner-sharing MO6 octahedra of high-spin M2+ ions. To gain insight into the magnetic properties of these phosphates, their spin exchange interactions were estimated by spin dimer analysis using tight binding calculations and by electronic band structure analysis using first principles density functional theory calculations. Three spin exchange interactions were found to be important for LiMPO4, namely, the intralayer superexchange J1, the intralayer super superexchange Jb along the b-direction, and the interlayer super-superexchange J2 along the b-direction. The magnetic ground state of LiMPO4 was determined in terms of these spin exchange interactions by calculating the total spin exchange interaction energy under the classical spin approximation. In the spin lattice of LiMPO4, the two-dimensional antiferromagnetic planes defined by the spin exchange J1 are antiferromagnetically coupled by the spin exchange J2, in agreement with available experimental data. PMID- 15792478 TI - Synthesis, characterization, physicochemical, and photophysical studies of redox switchable NIR dye derived from a ruthenium-dioxolene-porphyrin system. AB - Newly synthesized semi-quinone derivatives of the ruthenium polypyridyl, covalently linked to a porphyrin core, show very high epsilon values (59,000 83,500 M(-1)cm(-1)) for the absorption band in the near infrared (NIR) region of the spectrum. Further, complexes 1-4 show an interesting reversible electrochromic behavior as a function of the redox state of the coordinated dioxolene functionality, and a switching phenomenon between bleaching and the restoration of the NIR peak could be achieved electrochemically. Thus, complexes 1-4 could be ideal candidate materials for NIR-active electrochromic devices. Ultrafast studies on 1 and its mononuclear components, 5-(3,4-dihydroxyphenyl) 10,15,20-triphenyl-21H,23H-porphyrin (H2L1) and Ru(bpy)2(bsq)+, reveal that there is no electron or energy transfer from the porphyrin to the Ru(bpy)2sq+ (bpy is 2,2'-bipyridine and sq is the deprotonated species of a substituted semi-quinone fragment) fragment or vice versa in 1. The observed decrease in the luminescence quantum yield for 1 compared to that of H2L1 can be ascribed to the increased nonradiative pathway due to higher vibronic coupling because of the direct linkage of the metal center to the porphyrin moiety. PMID- 15792479 TI - Hydrothermal syntheses and crystal structures of two novel, hybrid materials based on secondary transition-metal-incorporated polyoxovanadate cluster backbones: [Cd(dien)2]2[(dien)CdAs8V13O41(H2O)].4H2O and [Cd(en)2]2[(en)2Cd2As8V12O40]. AB - Two new cadmium-substituted polyoxovanadates of [Cd(dien)2]2[(dien)CdAs8V13O41(H2O)].4H2O and [Cd(en)2]2[(en)2Cd2As8V12O40] (2), where dien is diethylenetriamine and en is ethylenediamine, have been hydrothermally synthesized and structurally characterized by elemental analyses, IR spectra, EPR spectra, XRD spectra, TG analyses, magnetic properties, and X-ray crystallography. Single-crystal X-ray diffraction analyses reveal that compounds 1 and 2 possess a discrete and 1D structure, respectively. The former contains an inorganic-organic hybrid monocadmium-substituted polyoxovanadate cluster unit that is derived from the well-known {As8V14O42} cluster, whereas the latter is composed of an inorganic-organic hybrid bicadmium-substituted polyoxovanadate cluster unit that is derived from the well-known {As8V12O40} cluster. PMID- 15792480 TI - Cluster oxalate complexes [M3(mu3-Q)(mu2-Q2)3(C2O4)3]2- and [Mo3(mu3-Q)(mu2 Q)3(C2O4)3(H2O)3]2- (M = Mo, W; Q = S, Se): mechanochemical synthesis and crystal structure. AB - Mechanochemical reaction of cluster coordination polymers 1infinity[M3Q7Br4] (M = Mo, W; Q = S, Se) with solid K2C2O4 leads to cluster core excision with the formation of anionic complexes [M3Q7(C2O4)3]2-. Extraction of the reaction mixture with water followed by crystallization gives crystalline K2[M3Q7(C2O4)3].0.5KBr.nH2O (M = Mo, Q = S, n = 3 (1); M = Mo, Q = Se, n = 4 (2); M = W, Q = S, n = 5 (3)). Cs2[Mo3S7(C2O4)3].0.5CsCl.3.5H2O (4) and (Et4N)1.5H0.5K{[Mo3S7(C2O4)3]Br}.2H2O (5) were also prepared. Close Q...Br contacts result in the formation of ionic triples {[M3Q7(C2O4)3](2)Br}5- in 1-4 and the 1:1 adduct {[Mo3S7(C2O4)3]Br}3- in 5. Treatment of 1 or 2 with PPh(3) leads to chalcogen abstraction with the formation of [Mo3(mu3-Q)(mu2 Q)3(C2O4)3(H2O)3]2-, isolated as (Ph4P)2[Mo3(mu3-S)(mu2-S)3(C2O4)3(H2O)3].11H2O (6) and (Ph4P2[Mo3(mu3-Se)(mu2-Se)3(C2O4)3(H2O)3].8.5H2O.0.5C2H5OH (7). All compounds were characterized by X-ray structure analysis. IR, Raman, electronic, and 77Se NMR spectra are also reported. Thermal decomposition of 1-3 was studied by thermogravimetry. PMID- 15792481 TI - An investigation on the role of the nature of sulfonate ancillary ligands on the strength and concentration dependence of the second-order NLO responses in CHCl3 of Zn(II) complexes with 4,4'-trans-NC5H4CH=CHC6H4NMe2 and 4,4'-trans,trans NC5H4CH=CH)2C6H4NMe2. AB - To evidentiate the role of the nature of sulfonate ancillary ligands on the value of the quadratic hyperpolarizability of Zn(II) complexes with stilbazole-like ligands, the second-order nonlinear optical (NLO) properties of [ZnY(2)(4,4' trans-NC5H4CH=CHC6H4NMe2)2] complexes (Y = CF3SO3, CH3SO3, or p-CH3C6H4SO3) are investigated. By working at relatively high concentrations (>3 x 10(-4) M), the positive effect of the triflate ligand remains unique while, with nonfluorinated sulfonate ligands, the second-order NLO response is comparable to that of the related complexes with acetate or trifluoroacetate as ancillary ligands. However, at dilutions higher than 10(-4) M, all of the sulfonate complexes reach huge quadratic hyperpolarizabilities because of solvolysis with the formation of cationic species such as [ZnY(4,4'-trans-NC5H4CH=CHC6H4NMe2)2]+, characterized by a large second-order NLO response. This view is supported by careful conductivity measurements. The same behavior occurs if 4,4'-trans-NC5H4CH=CHC6H4NMe2 is substituted by 4,4'-trans,trans-NC5H4(CH=CH)2C6H4NMe2. PMID- 15792482 TI - Synthesis, structural characterization, and photophysical properties of palladium and platinum(II) complexes containing 7,8-benzoquinolinate and various phosphine ligands. AB - A series of mononuclear cyclometalated benzo[h]quinolinate platinum and palladium(II) complexes with phosphine ligands, namely, [M(bzq)ClL] (L=PPh2H, Pt 1, Pd 2; PPh2CCPh, Pt 3, Pd 4), [Pt(bzq)(PPh2H)(PPh2CCPh)]ClO4 5, [Pt(bzq)(PPh2C(Ph)=C(H)PPh2)]ClO4 6, and [Pt(bzq)(CCPh)(PPh2CCPh)] (7a, 7b), were synthesized. The X-ray crystal structures of 1, 6.CH3COCH3.1/2CH3(CH2)4CH3, and 7b.CH3COCH3 have been determined. In 1, the metalated carbon atom and the P atom are mutually cis, whereas in 7b they are trans located. For complex 6, C and N are crystallographically indistinguishable. Reaction of [Pt(bzq)(mu-Cl)]2 with PPh2H and excess of NEt3 leads to the phosphide-bridge platinum dimer [Pt(bzq)(mu PPh2)]2 8 (X-ray). Moderate pi-pi intermolecular interactions and no evident Pt Pt interactions are found in 1, 7b, and in 8. All of the complexes exhibit absorption bands at high energy due to the intraligand transitions (1IL pi --> pi) and absorptions at lower energy which are attributed to MLCT (5d) pi --> pi (CLambdaN) transition. Platinum complexes show strong luminescence in both solid state and frozen solutions. The influence of the coligands on the photophysics of the platinum complexes has been examined by absorption and emission spectroscopy. PMID- 15792483 TI - Ligand-field excited states of metal hexacarbonyls. AB - Over 35 years ago, the low-lying bands in the absorption spectra of metal hexacarbonyls were assigned to ligand-field (LF) excitations. Recent time dependent density functional theory (TDDFT) calculations on M(CO)6 (M = Cr, Mo, W) are not in accord with this interpretation. Here we extend TDDFT calculations to the isoelectronic series V(CO)6-, Cr(CO)6, and Mn(CO)6+. By analyzing the trends in the energies of the various electronic excitations, we are able to fully assign the spectra of the complexes. In particular, we demonstrate that the LF excitation 1A1g -->1T1g is observed at 4.12 eV in the Mn(CO)6+ spectrum, but all LF features in the spectra of V(CO)6- and Cr(CO)6 are obscured by intense metal-to-ligand charge-transfer absorptions. Our results suggest that use of B3LYP as the exchange-correlation functional and inclusion of solvation effects through a continuum solvation model lead to the most accurate calculated transition energies. PMID- 15792484 TI - Synthesis and X-ray and neutron structures of Zr{(mu-H)2BC8H14}4. AB - The complex Zr(9-BBN)4 [9-BBN = (mu-H)2BC8H14] has been synthesized via the reaction of K(9-BBN) with ZrCl4 in diethyl ether. The structure of the title compound has been determined by X-ray and neutron single-crystal diffraction techniques. Each 9-BBN ligand is coordinated to the Zr atom via two B-H-Zr bridges, and these metal-ligand bonding interactions are further augmented by three prominent C-H...Zr agostic interactions. Average molecular parameters derived from the neutron analysis: Zr-H = 2.051(8) A, B-H = 1.286(7) A, Zr...B = 2.409(6) A, Zr-H-B = 87.7(4) degrees , H-Zr-H = 58.9(3) degrees . The Zr...H distances corresponding to the three C-H...Zr agostic interactions are 2.424(7), 2.663(8), and 2.551(7) A. The fourth potential C-H...Zr interaction has a Zr...H distance [3.146(7) A] that is too long to be considered in the agostic range. Single-crystal X-ray diffraction data were collected on an Enraf-Nonius Kappa CCD diffraction system, and neutron diffraction data were collected on the quasi-Laue diffractometer VIVALDI at the Institut Laue-Langevin; the final agreement factor for the neutron analysis is 6.52% for 2557 reflections with I > 2sigma(I). PMID- 15792485 TI - Oxidation of triarylphosphines and aryl methyl sulfides with hydrogen peroxide catalyzed by dioxovanadium(V) ion. AB - Although neither vanadium(V) ions nor hydrogen peroxide efficiently oxidize the title substrates, they do so in combination, with vanadium(V) as the catalyst in acidic aqueous acetonitrile. The kinetic data show that, of the two peroxovanadium species present, OV(O2)+ and OV(O2)2-, only the latter reacts at a detectable rate. This unanticipated result can be attributed to the weaker O-O and V-O bonds in the diperoxo complex. The rate constants for both series of substrates follow the Hammett correlation, with rhoP = -1.35 and rhoS = -0.83. To analyze properly the kinetic data for the Ar3P compounds, account must be taken of the protonation to Ar3PH+ in acidic solution. In retrospect, our earlier study [Abu-Omar, M. M.; Espenson, J. H. J. Am. Chem. Soc. 1995, 117, 272-280] of phosphine oxidation catalyzed by MeReO3 failed to do so, and the reaction constant must be corrected from the originally reported value to -1.56. PMID- 15792486 TI - Electrochemically informed synthesis and characterization of salts of the [Pt2(mu kappaAs,kappaC-C6H3-5-Me-2-AsPh2)4]+ lantern complex containing a Pt-Pt bond of order 1/2. AB - Detailed electrochemical studies in dichloromethane (0.1 M Bu4NPF6) on the oxidation of the half-lantern [Pt2(kappa2As,C-C6H3-5-Me-2-AsPh2)2(mu kappaAs,kappaC-C6H3-5-Me-2-AsPh2)2] (1) and full-lantern [Pt2(mu-kappaAs,kappaC C6H3-5-Me-2-AsPh2)4] (2) complexes reveal the presence of an exceptionally stable dinuclear Pt cation 2+. Thus, oxidation of 1 occurs on the voltammetric time scale via a ladder-square scheme to give 2+, whereas 2 is directly converted to 2+. Electrochemically informed chemical synthesis enabled the isolation of solid [2+][BF4-] to be achieved. Single-crystal X-ray structural analysis showed that 2+ also has a lantern structure but with a shorter separation between the Pt centers [2.7069(3) A (2+), 2.8955(4) A (2)]. EPR spectra of 2+ provide unequivocal evidence for axial symmetry of the complex and are noteworthy because of an exceptionally large, nearly isotropic hyperfine coupling constant of about 0.1 cm(-1). Spectroscopic data support the conclusion that the unpaired electron in the 2+ cation is distributed equally between the two Pt nuclei and imply that oxidation of 2 to 2+ leads to the establishment of the metal-to-metal hemibond. Results of extended Huckel molecular orbital and density functional calculations on 2 and 2+ lead to the conclusions that s, p, dz2 mixing of orbitals contributes to the large EPR Pt hyperfine coupling and also that the structural adjustments that occur upon removal of an electron from 2 are driven by the metal-metal bonding character present in 2+. PMID- 15792487 TI - Chemistry of ruthenium(II) monohydride and dihydride complexes containing pyridyl donor ligands including catalytic ketone H2-hydrogenation. AB - In this study we determine the changes to the properties of dihydride catalysts for ketone H2-hydrogenation by successively replacing the amine donors in the known dach complex RuH2(PPh3)2(dach) (2a), dach = 1,2-(R,R)-diaminocyclohexane, with one pyridyl group in the corresponding 2-(aminomethyl)pyridine (ampy) complexes RuH2(PPh3)2(ampy) (2b) and with two pyridyl groups in the complexes RuH2(PPh3)2(bipy) (2c) and RuH2(PPh3)2(phen) (2d). The ruthenium monohydride complex, (OC-6-54)-RuHCl(PPh3)2(ampy), (1b with Cl trans to H) was prepared by the addition of 1 equiv of ampy to RuHCl(PPh3)3 in THF. Treatment of the monohydride complex with K[BH(sec-Bu)3] in THF or KOtBu/H2 in toluene resulted in the formation of a mixture of at least two isomers of the highly reactive, air sensitive ruthenium dihydride complex 2b. One is the cis dihydride (OC-6-14)-2b or more simply c,t-2b with trans PPh3 groups and another is the cis dihydride c,c 2b (OC-6-42) that has PPh3 trans to H and PPh3 trans to N(pyridyl). The isomer c,c-2b slowly converts to c,t-2b in solution. The reaction of 1b with KOtBu under Ar results in the formation of a mixture that includes a complex with an imino ligand HN=CH-2-py while the same reaction under H2 leads to c,c-2b and then c,t 2b. The dach complex c,t-2a, reacts with ampy, 2,2'-bipyridine (bipy), and 1,10 phenanthroline (phen) in refluxing THF to form the substituted cis-dihydride complexes c,t-2b, (OC-6-13)-RuH2(PPh3)2(bipy) (c,t-2c with trans PPh3 groups) and (OC-6-13)-RuH2(PPh3)2(phen), c,t-2d, respectively. The dihydrides containing amino groups and cis-PPh3 groups, i.e., c,c-2a or c,c-2b, are active precatalysts for the H2-hydrogenation of acetophenone (neat or in benzene) under mild reaction conditions, whereas those with trans-PPh3 groups, c,t-2a and c,t-2b are much less active. The combination of ampy complex 1b and KOtBu also provides a catalyst in benzene that is more active than the corresponding dach system. The complexes without amino groups c,t-2c and c,t-2d are air-stable and inactive as hydrogenation catalysts under comparable conditions. The mechanism of hydrogenation of ketones catalyzed by isomers of 2a,b is thought to be similar and to proceed via a trans-dihydride complex, t,c-2a or t,c-2b, and an amido complex, neither of which are directly observed for the ampy complexes. The dihydride complex c,t-2b reacts with formic acid to give (OC-6-45) RuH(OCHO)(PPh3)2(ampy), 3b, with formate trans to hydride. The structures of 1b, c,t-2b, c,t-2c, and 3b have been determined by single-crystal X-ray diffraction. PMID- 15792488 TI - d-Electron-induced negative magnetoresistance of a pi-d interaction system based on a brominated-TTF donor. AB - A new pi-d interaction system (EDT-TTFBr2)2FeBr4 (EDT-TTFBr2 = 4,5-dibromo-4',5' ethylenedithiotetrathiafulvalene) and its nonmagnetic anion analogue (EDT TTFBr2)2GaBr4 based on a brominated TTF-type organic donor are investigated. The salts featured by quasi-1D pi-electronic systems are metallic with metal insulator transitions taking place at about 20 and 70 K for the FeBr4- and GaBr4- salts, respectively, where the low-temperature insulating state is associated with charge ordering or a Mott insulator followed by an antiferromagnetic transition at lower temperatures. The FeBr4- salt is featured with an antiferromagnetic transition of the anion d spins at a Neel temperature (TN) = 11 K, which is significantly high despite its long anion-anion Br-Br contact, suggesting the importance of the pi-d interaction in the magnetism. The surprisingly strong pi-d interaction, ca. -22.3 K estimated from the magnetization curve, evidences the usefulness of the chemical modification of the donor molecule with bromine substitution to achieve strong intermolecular interaction. The antiferromagnetic state of the anion d spins affects the transport of the conducting pi electrons through the strong pi-d interaction, as evidenced by the presence of a resistivity anomaly of the FeBr4- salt at TN. Below TN, the FeBr4- salt shows negative magnetoresistance that reaches -23% at the highest magnetic field investigated (B=15 T), whereas only a small positive magnetoresistance is observed in the pi-electron-only GaBr4- salt. The mechanism of the negative magnetoresistance is explained by the stabilization of the insulating state of the pi electrons by the periodic magnetic potential of the anion d spins in the FeBr4- salt, which is modified by applying the external magnetic field. PMID- 15792489 TI - Kinetics and mechanism of the mercury(II)-assisted hydrolysis of methyl iodide. AB - The kinetics and mechanism of the reaction of aqueous Hg(II) with methyl iodide have been investigated. The overall reaction is best described as Hg(II)-assisted hydrolysis, resulting in quantitative formation of methanol and, in the presence of excess methyl iodide, ultimately, HgI2 via the intermediate HgI+. The kinetics are biexponential when methyl iodide is in excess. At 25 degrees C, the acceleration provided by Hg2+ is 7.5 times greater than that caused by HgI+, while assistance of hydrolysis was not observed for HgI2. Thus, the reactions are not catalytic in Hg(II). The kinetics are consistent with an SN2-M+ mechanism involving electrophilic attack at iodide. As expected, methylation of mercury is not a reaction pathway; traces of methylmercury(II) are artifacts of the extraction/preconcentration procedure used for methylmercury analysis. PMID- 15792491 TI - Cyclic polyvanadates incorporating template transition metal cationic species: synthesis and structures of hexavanadate [PdV6O18]4-, octavanadate [Cu2V8O24]4-, and decavanadate [Ni4V10O30(OH)2(H2O)6]4-. AB - Three types of heteropolyvanadates, [(C2H5)4N]4[PdV6O18] (1), [(C2H5)4N]4[Cu2V8O24] (2), and [(C6H5)4P]4[Ni4V10O30(OH)2(H2O)6] (3), were synthesized through the reaction between the [VO3]- anion and metal template cations of Pd(II), Cu(II), and Ni(II). The X-ray crystal structures of 1 (a = 29.952(4) A, b = 12.911(2) A, and c = 13.678(2) A, orthorhombic, space group Pca2(1) with Z = 4), 2 (a = 13.740(1) A, b = 22.488(2) A, c = 18.505(2) A, and beta= 94.058(2) degrees , monoclinic, space group P2(1)/n with Z = 4), and 3 (a = 12.333(2) A, b = 16.208(4) A, c = 16.516(3) A, alpha = 112.438(3) degrees , beta = 94.735(3) degrees , and gamma = 104.749(3) degrees , triclinic, space group P with Z = 1) demonstrate that the metal cationic species induced cyclic [VO3](n-)n (n = 6, 8, 10) ring formation and the cations are incorporated in the rings themselves. In the metal inclusion products, the cyclic vanadates act as macrocyclic ligands, in which the metal cationic species act as the templates. The cyclic vanadate is composed of tetrahedral VO4 units that share corners and incorporates a metal cationic species in the center of the molecules. The bowl shaped complex 1 includes a Pd2+ cation that is coordinated by the oxygen donors of a boatlike hexavanadate ring. The diamagnetic complex 1 was characterized via 51V and 17O NMR spectroscopy. Complex 2 involves an octavanadate ring and two Cu2+, which are located on both sides of the mean plane as defined by the eight oxygen atoms that bridge the vanadium atoms. In the case of complex 3, the di-mu hydroxo-bridged Ni2+ dimer with capped Ni2+ aqua ions is formed by hydrolysis to form the decavanadate ring, in which two of the tetrahedral vanadate units are not bonded to the Ni2+ core but supported by hydrogen bonds through the aqua ligand in the capped Ni2+ cation. Complexes 1-3 in solution were clearly identified by their characteristic isotope patterns using ESI-MS studies. PMID- 15792490 TI - Mononuclear and dinuclear complexes of isoeilatin. AB - This work describes the synthesis and characterization of mononuclear and dinuclear Ru(II) and Os(II) complexes based on the symmetrical bridging ligand isoeilatin (1). The crystal structure of 1.[HCl]2 consists of layers of tightly pi-stacked molecules of the biprotonated isoeilatin. The mononuclear complexes [Ru(bpy)2(ieil)]2+ (2(2+)) and [Os(bpy)2(ieil)]2+ (3(2+)) form discrete dimers in solution held together by face-selective pi-stacking interactions via the isoeilatin ligand. Coordination of a second metal fragment does not hinder the pi stacking completely, as demonstrated by the concentration dependence of the 1H NMR spectra of the dinuclear complexes [{Ru(bpy)2}2{mu-ieil}]4+ (4(4+)), [{Os(bpy)2}2{mu-ieil}]4+ (5(4+)), and [{Ru(bpy)2}{mu-ieil}{Os(bpy)2}]4+ (6(4+)) and supported by the solid-state structure of meso-4.[Cl]4. The bridging isoeilatin ligand conserves its planarity even upon coordination of a second metal fragment, as demonstrated in the solid-state structures of meso-4.[Cl]4, meso-4.[PF6]4, and meso-5.[PF6]4. All of the dinuclear complexes exhibit a preference (3/2-3/1) for the formation of the heterochiral as opposed to the homochiral diastereoisomer. Absorption spectra of the mononuclear complexes feature a low-lying dpi(M) --> pi*iel MLCT band around 600 nm that shifts to beyond 700 nm upon coordination of a second metal fragment. Cyclic and square wave voltammetry measurements of the complexes exhibit two isoeilatin-based reduction waves that are substantially anodically shifted compared to [M(bpy)3]2+ (M = Ru, Os). Luminescence spectra, quantum yields, and lifetime measurements at room temperature and at 77 K demonstrate that the complexes exhibit 3MLCT emission that occurs in the IR region between 950 and 1300 nm. Both the electrochemical and photophysical data are consistent with the low-lying pi orbital of the isoeilatin ligand. The dinuclear complexes exhibit two reversible, well-resolved, metal-centered oxidation waves, despite the chemical equivalence of the two metal centers, indicating a significant metal-metal interaction mediated by the bridging isoeilatin ligand. PMID- 15792492 TI - Coordination modes between copper(II) and N-acetylneuraminic (sialic) acid from a 2D-simulation analysis of EPR spectra. Implications for copper mediation of sialoglycoconjugate chemistry relevant to human biology. AB - The equilibrium distribution of species formed between Cu(II) and N acetylneuraminic (sialic) acid (I, LH) at 298 K has been determined using a two dimensional (2D) simulation analysis of electron paramagnetic resonance (EPR) spectra. In acidic solutions (pH values < 4), the major species present are Cu(2+), [CuL]+ [logbeta = 1.64(4)], and [CuL2] [logbeta = 2.77(5)]. At intermediate pH values (4.0 < pH < 7.5), [CuL2H-1]- [logbeta = -2.72(7)] and two isomers of [CuLH-1] [logbeta (overall) = -3.37(2)] are present. At alkaline pH values (7.5 < pH < 11), the major species present is [CuL2H-2]2-, modeled as three isomers with unique giso and Aiso values [logbeta (overall) = -8.68(3)]. Two further species ([CuLH-3]2- and [CuL2H-3]3-) appear at pH values > 11. It is proposed that [CuL]+ most likely features I coordinated via the deprotonated carboxylic acid group (O1) and the endocyclic oxygen atom (OR) forming a five membered chelate ring. Select Cu(II)-I species of the form [CuLH-1] may feature I acting as a dianionic tridentate chelate, via oxygen atoms derived from O1, OR, and one deprotonated hydroxy group (O7 or O8) from the glycerol tail. Alternatively, I may coordinate Cu(II) in a bidentate fashion as the tert-2 hydroxycarboxylato (O1,O2) dianion. Spectra predicted for Cu(II)-I complexes in which I is coordinated in either a O1,OR {I1-} or O1,O2 {I2-} bidentate fashion {e.g., [CuL]+ (O1,O R), [CuL2] (bis-O1,O R), [CuLH-1] (isomer: O1, O2), [CuL2H-1] (O1, O R; O1, O2), and [CuL2H-2]2- (isomer: bis-O1, O2)} have "irregular" EPR spectra that are ascribed to the existence of Cu(II)-I(monomer) <==> Cu(II) I(polymer) equilibria. The formation of polymeric Cu(II)-I species will be favored in these complexes because the glycerol-derived hydroxyl groups at the complex periphery (O, 7O, 8O9) are available for further Cu(II) binding. The presence of polymeric Cu(II)-I species is supported by EPR spectral data from solutions of Cu(II) and the homopolymer of I, colominic acid (Ipoly). Conversely, spectra predicted for Cu(II)-I complexes where I is coordinated in a {I2-} tridentate {e.g., [CuLH-1] (isomer: O1, O R, O7, or O8) and [CuL2H-2]2- (isomer: bis-O1,O R,O7, or O8)} or tetradentate fashion {I3-} {e.g., [CuLH-3]2- (O1, O R, O, 8O9)} are typical for mononuclear tetragonally elongated Cu(II) octahedra. In this latter series of complexes, the tendency toward the formation of polymeric Cu(II)-I analogues is small because the polydentate I effectively wraps up the mononuclear Cu(II) center. This work shows that Cu(II) could potentially mediate the chemistry of sialoglycoconjugate-containing proteins in human biology, such as the sialylated amyloid precursor protein of relevance to Alzheimer's disease. PMID- 15792493 TI - Flexible ligands and structural diversity: isomerism in Cd(NO3)2 coordination polymers. AB - The ligands 1,4-bis(2-pyridylmethylsulfanylmethyl)benzene (L1) and 2,5-bis(2 pyridylmethylsulfanylmethyl)pyrazine (L2) were treated with Cd(NO3)2.4H2O in metal-to-ligand ratios of 1:1 and 2:1, respectively; L2 was also treated with CdCl2.2.5H2O in a 2:1 ratio. All products were found to be coordination polymers. The crystal structures of {[Cd(L1)(NO3)2].CH2Cl2}infinity (1a), {[Cd(L1)(NO3)2].4/3CH3CN}infinity (1b), {[Cd2(L2)(NO3)4].2CH3CN}infinity (2.2CH3CN), and {[Cd2(L2)Cl4].2CH2Cl2}infinity (3.2CH2Cl2) were determined. Compounds 1a and 1b were found to be conformational supramolecular isomers. The structure of 1b displayed topological isomerism with two isomeric polymer chains, 1b(1) and 1b(2), in the one crystal forming a single supramolecular array. The structure of 2.2CH(3)CN showed Cd2(L2) units linked together by nitrates bridging between the Cd(II) centers in a mode previously not seen in Cd(II) compounds. The overall structure of 3.2CH2Cl2 was found to be similar to that of 2.2CH3CN despite the presence of different anions and solvent molecules. Powder X-ray diffraction was used to investigate the nature of bulk preparations of compounds 1-3. PMID- 15792494 TI - Synthesis, structure, and large optical limiting effect of the first coordination polymeric cluster based on an {I@[AgI(inh)]6} hexagram block. AB - In this paper, treatment of N-(isonicotinoyl)-N'-nicotinoylhydrazine (inh) and AgI with excess KI afforded a unique coordination polymeric cluster {[AgI(inh)]6(KI)}n (1) with the hexagram cluster units centered by mu6-I. In the polymer these hexagram units are parallel to the ab plane and are linked by separated K+ centers through inh. Polymer 1 represents the first example of coinstantaneous cation-anion-induced supramolecular self-assembly with nanoscale inner cavities. The polymer's third-order nonlinear optical (NLO) properties were determined by the Z-scan technique in DMF solution. The results show that the polymer has strong third-order optical nonlinearities. The nonlinear absorptive index a2 and refractive index n2 are calculated to be 1.044 x 10(-9) mW(-1) and 2.827 x 10(-11) esu, respectively. The values are comparable to those of the reported cluster polymers. The optical limiting experiments show that the present cluster exhibits a large optical limiting capacity. The value of the limiting threshold was measured as 0.53 J cm(-2) from the optical limiting experimental data. This value is three times better than 1.6 J cm(-2) of C60. This paper also gives a summary and comparison on the optical limiting properties of oligomeric and polymeric clusters. PMID- 15792495 TI - New oxorhenium(V) compound for catalyzed oxygen atom transfer from picoline N oxide to triarylphosphines. AB - The synthesis and characterization of a new oxorhenium(V) compound is reported; it is [MeReO(edt)(bpym)], 8, where edt = 1,2-ethanedithiolate and bpym = 2,2' bipyrimidine. Compound 8 was characterized by NMR spectroscopy and single-crystal X-ray analysis. It exists as a six-coordinate Re(V) compound comparable to the previously known [MeReO(edt)(bpy)] and [MeReO(mtp)(bpy)]. Compound 8 catalyzes the oxygen-atom-transfer reaction PicO + PZ3 --> Pic + Z3PO, whereas the other two do not. The kinetics of this reaction with catalyst 8 follows the rate law d[PicO]/dt = k[8][PicO]/(1 + c[PZ3]). With different phosphines, the rate law has the same k value, 4.17 L mol(-1) s(-1), but different c values. For tritolylphosphine, c = 67.5 L mol(-1) in benzene at 25 degrees C. A mechanism has been proposed to account for these findings. The data establish that an open coordination site on rhenium is necessary for oxygen-atom-transfer reactions. PMID- 15792496 TI - Interplay between DNA replication and recombination in prokaryotes. AB - The processes of DNA replication and recombination are intertwined at many different levels. In diverse systems, extensive DNA replication can be triggered by genetic recombination, with assembly of a replication complex onto a D-loop recombination intermediate. This and related pathways of replisome assembly allow the completion of DNA replication when forks initiated at a conventional replication origin fail before completing replication of the genome. In addition, the repair of double-strand breaks or gaps by homologous recombination requires at least limited DNA replication to replace the missing information. An intricate interplay between replication and recombination is also evident during the termination of bacterial DNA replication and during the induction of the bacterial SOS response to DNA damage. PMID- 15792498 TI - Psychometric properties of the Flemish translation of the NEECHAM Confusion Scale. AB - BACKGROUND: Determination of a patient's cognitive status by use of a valid and reliable screening instrument is of major importance as early recognition and accurate diagnosis of delirium is necessary for effective management. This study determined the reliability, validity and diagnostic value of the Flemish translation of the NEECHAM Confusion Scale. METHODS: A sample of 54 elderly hip fracture patients with a mean age of 80.9 years (SD = 7.85) were included. To test the psychometric properties of the NEECHAM Confusion Scale, performance on the NEECHAM was compared to the Confusion Assessment Method (CAM) and the Mini Mental State Examination (MMSE), by using aggregated data based on 5 data collection measurement points (repeated measures). The CAM and MMSE served as gold standards. RESULTS: The alpha coefficient for the total NEECHAM score was high (0.88). Principal components analysis yielded a two-component solution accounting for 70.8% of the total variance. High correlations were found between the total NEECHAM scores and total MMSE (0.75) and total CAM severity scores ( 0.73), respectively. Diagnostic values using the CAM algorithm as gold standard showed 76.9% sensitivity, 64.6% specificity, 13.5% positive and 97.5% negative predictive values, respectively. CONCLUSION: This validation of the Flemish version of the NEECHAM Confusion Scale adds to previous evidence suggesting that this scale holds promise as a valuable screening instrument for delirium in clinical practice. Further validation studies in diverse clinical populations; however, are needed. PMID- 15792500 TI - In memorium: Herman p. Schwan [1915-2005]. AB - Herman P. Schwan [1915-2005] was a distinguished scientist and engineer, and a founding father of the field of biomedical engineering. A man of integrity, Schwan influenced the lives of many, including his wife and children, and his many students and colleagues. Active in science until nearly the end of his life, he will be very much missed by his family and many colleagues. PMID- 15792499 TI - Coronary flow reserve in stress-echo lab. From pathophysiologic toy to diagnostic tool. AB - The assessment of coronary flow reserve by transthoracic echocardiography has recently been introduced into clinical practice with gratifying results for the diagnosis of left anterior descending artery disease simultaneously reported by several independent laboratories. This technological novelty is changing the practice of stress echo for 3 main reasons. First, adding coronary flow reserve to regional wall motion allows us to have - in the same sitting - high specificity (regional wall motion) and a high sensitivity (coronary flow reserve) diagnostic marker, with an obvious improvement in overall diagnostic accuracy. Second, the technicalities of coronary flow reserve shift the balance of stress choice in favour of vasodilators, which are a more robust hyperemic stress and are substantially easier to perform with dual imaging than dobutamine or exercise. Third, the coronary flow reserve adds a quantitative support to the exquisitely qualitative assessment of wall motion analysis, thereby facilitating the communication of stress echo results to the cardiological world outside the echo lab. The next challenges involve the need to expand the exploration of coronary flow reserve to the right and circumflex coronary artery and to prove the additional prognostic value - if any - of coronary flow reserve over regional wall motion analysis, which remains the cornerstone of clinically-driven diagnosis in the stress echo lab. PMID- 15792502 TI - Prediction of cerebral vasospasm in patients presenting with aneurysmal subarachnoid hemorrhage: a review. AB - OBJECTIVE: Cerebral vasospasm is a devastating medical complication of aneurysmal subarachnoid hemorrhage (SAH). It is associated with high morbidity and mortality rates, even after the aneurysm has been treated. A substantial amount of experimental and clinical research has been conducted in an effort to predict and prevent its occurrence. This research has contributed to significant advances in the understanding of the mechanisms leading to cerebral vasospasm. The ability to accurately and consistently predict the onset of cerebral vasospasm, however, has been challenging. This topic review describes the various methodologies and approaches that have been studied in an effort to predict the occurrence of cerebral vasospasm in patients presenting with SAH. METHODS: The English-language literature on the prediction of cerebral vasospasm after aneurysmal SAH was reviewed using the MEDLINE PubMed (1966-present) database. RESULTS: The risk factors, diagnostic imaging, bedside monitoring approaches, and pathological markers that have been evaluated to predict the occurrence of cerebral vasospasm after SAH are presented. CONCLUSION: To date, a large blood burden is the only consistently demonstrated risk factor for the prediction of cerebral vasospasm after SAH. Because vasospasm is such a multifactorial problem, attempts to predict its occurrence will probably require several different approaches and methodologies, as is done at present. Future improvements in the prevention of cerebral vasospasm from aneurysmal SAH will most likely require advances in our understanding of its pathophysiology and our ability to predict its onset. PMID- 15792501 TI - A qualitative study to identify community structures for management of severe malaria: a basis for introducing rectal artesunate in the under five years children in Nakonde District of Zambia. AB - BACKGROUND: Malaria is a serious illness among children aged 5 years and below in Zambia, which carries with it many adverse effects including anemia and high parasites exposure that lead to infant and childhood mortality. Due to poor accessibility to modern health facilities, malaria is normally managed at home using indigenous and cosmopolitan medicines. In view of problems and implications associated with management of severe malaria at home, rectal artesunate is being proposed as a first aid drug to slow down multiplication of parasites in children before accessing appropriate treatment. METHODS: A qualitative study using standardised in-depth and Focus Group Discussions (FGDs) guides to collect information from four (4) villages in Nakonde district, was conducted between February and March 2004. The guides were administered on 29 key informants living in the community and those whose children were admitted in the health facility. Participants in the 12 FGDs came from the 4 participating villages. Participants and key informants were fathers, younger and older mothers including grandmothers and other influential people at household level. Others were traditional healers, headmen, village secretaries, traditional birth attendants, church leaders and blacksmiths. FGDs and interview transcriptions were coded to identify common themes that were related to recognition, classification and naming of malaria illness, care-seeking behaviour and community treatment practices for severe malaria. RESULTS: Parental prior knowledge of the disease was important as the majority of informants (23 out of 29) and participants (69 out of 97) mentioned four combined symptoms that were used to recognise severe malaria. The symptoms were excessive body hotness, convulsions, vomiting yellow things and bulging of the fontanelle. On the other hand, all informants mentioned two or more of symptoms associated with severe malaria. In all 12 FGDs, participants reported that treatment of severe malaria commenced with the family and moved into the community as the illness progressed. Although treatment of severe diarrheal effects, were common among the Winamwanga, no rectal medicines to treat severe malaria were identified. Apart from the anti-malarial fansidar, which was mentioned by 23 in IDIs and 40 in FGDs, participants and informants also frequently mentioned indigenous medicines provided by healers and other respectable herbalists for repelling evil spirits, once a child had severe malaria. Mothers were the important arms for administration of ant-malarial drugs in the villages. Referrals began with healers to CHWs, where no CHWs existed healers directly referred sick children to the health facility. CONCLUSION: Our findings showed that there is a precedent for rectal application of traditional medicine for childhood illness. Therefore rectal artesunate may be a well received intervention in Nakonde District, provided effective sensitisation, to mothers and CHWs is given which will strengthen the health care delivery system at community level. PMID- 15792503 TI - Familial aggregation of both aortic and cerebral aneurysms: evidence for a common genetic basis in a subset of families. AB - OBJECTIVE: Although previous reports have described patients with both cerebral and aortic aneurysms, any association was believed to be coincidental. In this study, we provide evidence that aortic and cerebral aneurysm formation may share a common genetic predisposition in some families. METHODS: A prospective enrollment of consecutive patients treated for saccular cerebral aneurysm by a single surgeon was constructed. Medical and family histories were obtained. Familial syndromes were identified when two or more first-degree relatives had diagnoses of cerebral or aortic aneurysm. Pedigrees were constructed, and asymptomatic relatives were screened. RESULTS: In 2.5 years, 274 patients were enrolled from 322 eligible patients (85%). A family history of aortic aneurysm was noted in 29 patients (10.5%). These patients were older, included more males, and were overwhelmingly Caucasian. Statistically significant demographic differences were noted when these families were compared with families affected by cerebral aneurysm only. A total of 110 affected family members were identified (average, 3.8 per family; range, 2-12); 59 had cerebral aneurysms, and 51 had aortic aneurysms. In some families, the association may have been coincidental. But in several families, pedigree analyses showed an inheritance pattern likely to represent autosomal dominance with variable penetrance. In addition, both cerebral and aortic phenotypes could be inherited from a parent with an aortic aneurysm, further evidence for a common genetic basis. CONCLUSION: This study, which represents the largest and most complete characterization of families affected by both cerebral and aortic aneurysms, provides evidence that a single gene defect may lead to the development of either lesion. PMID- 15792504 TI - Clinical features of aneurysms of the posterior cerebral artery: a 15-year experience with 21 cases. AB - OBJECTIVE: To investigate the characteristic clinical behavior and develop guidelines for the clinical management of posterior cerebral artery aneurysms, we reviewed 21 cases treated during a 15-year period. METHODS: Between 1988 and 2002, we treated 21 patients (10 male, 11 female; mean age, 49.8 yr) with posterior cerebral artery aneurysms at Kumamoto University Hospital and its affiliated hospitals. Data relating to the clinical, radiological, and surgical approaches were analyzed. RESULTS: There were 20 saccular aneurysms, 6 of which were giant or large aneurysms, and 1 dissecting aneurysm. Of the 21 patients, 15 presented with subarachnoid hemorrhage; in 3 patients, an unruptured saccular aneurysm was found incidentally during an evaluation for other cerebral pathological conditions. Two patients presented with progressive homonymous hemianopsia because of the mass effect of the aneurysm, and 1 patient experienced the sudden onset of homonymous hemianopsia because of thrombosis of the aneurysm and afferent artery. Open or endovascular surgery was performed in 19 patients: 14 made a good recovery, 2 had a moderate disability because of angiospasm or infarction after aneurysm trapping, 1 had a severe disability because of angiospasm and cerebral contusion, and 2 died because of severe angiospasm. Of 2 conservatively treated patients, 1 made a good recovery but the other died as a result of rebleeding. CONCLUSION: Posterior cerebral artery aneurysms have specific clinical characteristics compared with aneurysms located elsewhere. The aneurysmal site and size and the surgical technique did not significantly influence treatment outcomes. PMID- 15792505 TI - Factors associated with neurological outcome and lesion progression in traumatic subarachnoid hemorrhage patients. AB - OBJECTIVE: Traumatic subarachnoid hemorrhage (tSAH) is a frequent finding after closed-head injuries, and its presence is a powerful factor associated with poor outcome. The exact mechanism linking tSAH and an adverse outcome is poorly understood. The aim of this study was to identify the factors that may predict outcomes and changes in the computed tomographic (CT) scans of lesions in a selected population of tSAH patients. METHODS: We evaluated 141 patients admitted consecutively from January 1, 1997, to January 31, 1999, with a CT diagnosis of tSAH. The admission and "worst" CT scans were recorded. CT scan changes were reported as "significant CT progression" (changes in the Marshall classification) or "any CT progression." The amount of subarachnoid blood was recorded using a modified Fisher classification. Outcome was assessed at 6 months after injury with the Glasgow Outcome Scale. RESULTS: Twenty-eight patients (19.9%) had an unfavorable Glasgow Outcome Scale outcome. In the univariate analysis, prognosis was significantly related to age, admission Glasgow Coma Scale score, Marshall CT classification score at admission and on the worst CT scan, amount of tSAH, and volume of the associated brain contusions. From multivariate analysis, the only factors independently related to outcome were the Glasgow Coma Scale score (P < 0.01) and size of the tSAH at admission (P < 0.001). Thirty-four patients (24.1%) had significant CT lesion progression, and 66 patients (46.8%) had some lesion progression. Patients having significant progression of the lesion had a higher risk of an unfavorable outcome (32 versus 10%; P = 0.004). Unadjusted factors predicting CT progression were the Glasgow Coma Scale score at admission, the Marshall classification at admission, the amount of subarachnoid blood, and the presence or volume of associated brain contusions at admission. Independent factors associated with significant CT progression were the amount of tSAH (P < 0.001) and the presence or volume of brain contusions at admission (P < 0.001). CONCLUSION: The outcome of patients with tSAH at admission is related in a logistic regression analysis to the admission Glasgow Coma Scale score and to the amount of subarachnoid blood. These patients also have a significant risk of CT progression. The amount of subarachnoid blood and the presence of associated parenchymal damage are powerful independent factors associated with CT progression, thus linking poor outcomes and CT changes. PMID- 15792506 TI - Factors associated with outcome after hemicraniectomy for large middle cerebral artery territory infarction. AB - OBJECTIVE: Large or malignant middle cerebral artery infarction is associated with a high mortality rate. Hemicraniectomy reduces intracranial pressure and has been demonstrated to increase survival. Questions remain regarding selection of patients for surgery and functional outcomes. METHODS: We retrospectively reviewed the charts, operative reports, and images of patients who underwent hemicraniectomy for cerebral infarction at our institution between 1990 and 2002. Stroke volume and craniectomy area image analyses were performed with National Institutes of Health Image J software. Short-term outcome was measured with the Glasgow Outcome Scale. The requirement for second surgical decompression was also used as a measure of short-term outcome. Outcome 1 year after stroke was assessed with mailed self-report questionnaires to patients or providers with answers yielding information pertinent to the Barthel Index for physical disability, the Reintegration to Normal Living Index, and the Zung Depression Index. Univariate and multivariate logistic regression analyses were used to consider factors associated with outcome. RESULTS: Thirty-eight patients with large hemispheric infarcts (average volume, 407 cm3) were treated with hemicraniectomy during the study period. Thirty-two patients survived more than 1 year after surgery. Twenty patients responded to a questionnaire scoring late physical disability (Barthel Index), quality of life (Reintegration to Normal Living Index), and depression (Zung Depression Index). The average Barthel Index score was 67. Barthel Index score and ability to walk were strongly correlated with age but not time to surgery, volume of infarction, or craniectomy size. Patients exhibited moderate to severe decrements in quality of life and increased incidence of depression across all age groups. Reoperation, an indicator for early operative failure, was required in six patients. These patients were significantly younger and had significantly higher volumes of cerebral infarction. CONCLUSION: Hemicraniectomy is life-saving treatment for large middle cerebral artery infarction. Good functional outcomes can be obtained but are less likely in older patients. Younger patients with large-volume strokes may benefit from multiple decompressions, if necessary. PMID- 15792507 TI - Clinical experience with the intraparenchymal intracranial pressure monitoring Codman MicroSensor system. AB - OBJECTIVE: Our main objective was to study the reliability of the Codman MicroSensor (CMS), used for intracranial pressure (ICP) measurements, as it is used in a clinical setting. In particular, the drift from zero was studied. METHODS: The investigation is a prospective study of 128 patients with a need for neurointensive care who have been treated for various diseases. The patients received an intraparenchymal CMS device, and the zero drift was measured at explantation of the sensor. In another 22 patients, the ICP was recorded simultaneously from a ventriculostomy and a CMS, and the values were compared. The general data of complications and pitfalls are collected from close to 1000 CMS devices implanted. RESULTS: The CMS was used, on average, 7.2 +/- 0.4 days per patient. The total time of ICP measurement was 20,040 hours, resulting in at least 7.2 x 10(7) measuring values displayed. The drift from zero was 0.9 +/- 0.2 mm Hg, and no correlation with duration of use was found (P = 0.9, r = 0.002). There was a good correlation between ICP measured by CMS and by ventriculostomy (P < 0.0001, r = 0.79). The average ICP measured with the ventriculostomy was 18.3 +/- 0.3 mm Hg, and with the CMS, it was 19.0 +/- 0.2 mm Hg. A few minor hematomas were identified, and no infections directly connected to the device were observed. Some pitfalls in handling and problems during magnetic resonance imaging investigations are discussed. CONCLUSION: In our hands, the CMS device is reliable and easy to use. The ICP recordings are stable over time, and there is only a minor zero drift. The device is today our standard method for ICP measurements. PMID- 15792508 TI - Radiosurgery for nonfunctioning pituitary adenomas. AB - OBJECTIVE: We evaluated the effectiveness of gamma knife radiosurgery in the treatment of nonfunctioning pituitary adenomas. METHODS: Between January 1994 and December 1999, we treated 34 patients with nonfunctioning pituitary adenomas. Thirty-one of these patients were followed for more than 30 months. Their mean age was 52.9 years. All patients underwent resection before radiosurgery. In four patients, treatment was performed with staged radiosurgery. The treatment volume was 0.7 to 36.2 cm3 (median, 2.5 cm3). The treatment dose ranged from 8 to 20 Gy (median, 14.0 Gy) to the tumor margin. In 15 patients (48.4%), the tumor either compressed or was attached to the optic apparatus. The maximum dose to the optic apparatus was from 2 to 11 Gy (median, 8 Gy). RESULTS: Patients were followed for 30 to 108 months (median, 59.8 mo). The tumor size decreased in 18 patients (58.1%), remained unchanged in 9 patients (29.0%), and increased in four patients (12.9%). The 5-year actual tumor growth control rate was 93%. Among patients with tumor growth, two cases were secondary to cyst formation. Two patients (6.5%) required adrenal and thyroid hormonal replacement during the follow-up period after radiosurgery because of radiation-induced endocrinopathy. None of the patients sustained new cranial nerve deficits, which included optic neuropathy. CONCLUSION: In this series, radiosurgery had a high tumor growth control rate during the long-term follow-up period. Furthermore, we observed a low morbidity rate, with endocrinopathies and optic neuropathies. This low rate included even patients in whom the tumor compressed or was attached to the optic apparatus. We emphasize the necessity of long-term follow-up to evaluate late complications. PMID- 15792509 TI - Surgical outcome in rheumatoid Ranawat Class IIIb myelopathy. AB - OBJECTIVE: Rheumatoid arthritis frequently affects the craniovertebral junction (CVJ) and may lead to severe neck pain, quadriparesis, and respiratory dysfunction. Surgery in rheumatoid nonambulatory (Ranawat Class IIIb) patients carries a significant risk. This study presents the surgical outcome of Class IIIb patients with CVJ rheumatoid myelopathy and reviews the literature. METHODS: One hundred twelve consecutive patients with rheumatoid cervical myelopathy underwent surgical decompression and stabilization. Thirty-two of the patients (mean age, 66.81 +/- 10.25 yr) with CVJ rheumatoid arthritis were in Class IIIb, and all had atlantoaxial subluxation. A halo brace was applied before surgery and continued during surgery. Eleven patients with reducible atlantoaxial subluxation underwent direct posterior fusion. Twenty-one patients with fixed atlantoaxial subluxation underwent transoral decompression and then posterior fusion while they were under anesthesia. RESULTS: At a mean follow-up of 39 months, four patients improved to Class II and 14 improved to Class IIIa, whereas six remained in Class IIIb. Neck pain was relieved in all patients. There was one perioperative death after transoral surgery (posterior fusion not done), and seven other patients died subsequently of causes unrelated to surgery. The morbidity of surgery included construct failure, cerebrospinal fluid leak, superficial wound or graft donor site infection, transient dysphagia, and lung infection. CONCLUSION: A large subset of patients with CVJ rheumatoid myelopathy may reach Class IIIb. These patients have unique management considerations. Surgery (despite high morbidity) often remains the best therapeutic option available to them. Improvement of even one grade in their Ranawat score from Class IIIb to Class IIIa brought about by surgery confers on them a significant benefit in terms of their quality of life and survival. PMID- 15792510 TI - Correlation of cerebrospinal fluid flow dynamics and headache in Chiari I malformation. AB - OBJECTIVE: The management of patients with a Chiari I malformation who present with headaches alone remains unclear. We studied the cerebrospinal fluid (CSF) flow dynamics of Chiari I malformation patients presenting with headaches alone so as to identify headache types that are associated with CSF flow obstruction versus those that may be unrelated to Chiari I malformations. METHODS: Preoperative cine phase-contrast magnetic resonance imaging of the craniocervical junction was prospectively performed in 33 patients presenting with headaches alone and a Chiari I malformation (tonsillar ectopia >5 mm below the foramen magnum). Headaches were classified as frontal, occipital, or generalized. CSF flow dynamics were then prospectively compared with presenting symptomatology. A subgroup of 17 patients underwent surgical decompression of the Chiari I malformations. RESULTS: Patients with frontal or generalized headaches were 10 fold less likely to demonstrate obstructed CSF flow (odds ratio, 0.10; 95% confidence interval, 0.02-0.52) and 8-fold less likely to have tonsillar descent greater than 7 mm (odds ratio, 0.12; 95% confidence interval, 0.03-0.62) compared with patients with occipital headaches. Adjusting for degree of tonsillar herniation in multivariate analysis, frontal and generalized headaches remained independently associated with nonobstructed CSF flow pathological findings, whereas occipital headaches remained associated with obstructed CSF flow independent of tonsil location (odds ratio, 5.84; 95% confidence interval, 1.01 34.28). In the surgical group, all patients with obstructed CSF flow did well compared with the group with normal flow, regardless of headache location. CONCLUSION: Regardless of the degree of tonsillar ectopia, occipital headaches were strongly associated with hindbrain CSF flow abnormalities, whereas frontal and generalized headaches were not. Normal magnetic resonance imaging-cine CSF flow in the setting of a Chiari I malformation and frontal headaches alone suggests that frontal headaches are not pathologically or causatively associated with the Chiari I malformation in the vast majority of patients. Frontal headaches with obstructed flow may respond to surgery. PMID- 15792511 TI - Risk factors for hemorrhage during microelectrode-guided deep brain stimulator implantation for movement disorders. AB - OBJECTIVE: Although hemorrhage is a well-known complication of microelectrode guided deep brain stimulation (DBS) surgery, risk factors for the development of hemorrhage have not been well defined. We analyzed the risk factors for symptomatic and asymptomatic hemorrhage in a large series of DBS implantations into the subthalamic nucleus, ventrolateral thalamus, and internal globus pallidus. METHODS: All DBS procedures performed by a single surgeon at our institution between June 1998 and May 2004 were included in this study. All patients had postoperative imaging (magnetic resonance imaging or computed tomography) 4 to 24 hours after surgery. Hematomas were noted and scored as symptomatic or asymptomatic. Statistical correlation of factors affecting risk of hematoma formation was performed by use of logistic regression analysis. RESULTS: The total number of lead implantations was 481. There were 6 symptomatic hematomas and 10 asymptomatic hematomas. Three of the symptomatic hematomas resulted in permanent new neurological deficit. The risk of hematoma (of any type) per lead implantation was 3.3%, whereas the risk of permanent deficit from hematoma was 0.6%. Patients who developed hematomas had a slightly greater number of microelectrode recording penetrations than patients who did not have hematomas, but this difference did not reach statistical significance. There was not a statistically significant relationship between risk of hematoma and patient age or diagnosis. There was a significant effect of brain target (P = 0.001), with only 1 hemorrhage detected after thalamic DBS. CONCLUSION: DBS is generally safe, with only 0.6% of implantations associated with permanent neurological deficit. The incremental risk of successive serial microelectrode penetrations is small. PMID- 15792512 TI - Surgical site infections after pediatric intracranial surgery for craniofacial malformations: frequency and risk factors. AB - OBJECTIVE: Surgical site infections (SSIs) after pediatric intracranial surgery for craniofacial malformations are relatively common and potentially serious nosocomial infections. Despite this, few studies have been published on this topic. We performed a retrospective study to determine the incidence of and factors associated with SSIs after intracranial surgery for craniofacial malformations at a single multidisciplinary craniofacial center during a 6-year period. METHODS: Retrospective chart review was performed on 254 patients who underwent 281 intracranial procedures during a 6-year period. Patients with SSIs (cases) were compared with those without SSIs (controls). National Nosocomial Infection Surveillance System criteria were used to identify cases of SSI. RESULTS: SSIs occurred in 9 (3.2%) of 281 intracranial procedures that took place during the study period. Factors associated with an SSI included a complicated diagnosis comprising a diagnosis of syndromic craniosynostosis, frontonasal dysplasia, or oblique facial cleft (odds ratio [OR], 13.0; 95% confidence interval [CI], 2.6-64.4); duration of surgery longer than 426 minutes (OR, 12.1; 95% CI, 2.4-59.9); closure of skin under tension (OR, 12.5; 95% CI, 3.0-52.6); use of bovine pericardium (OR and 95% CI undefined); more than four surgeons present during surgery (OR, 6.3; 95% CI, 1.2-32); pediatric intensive care unit stay longer than 2 days (OR, 10.8; 95% CI, 2.2-53.3); and use of a ventilator after surgery (OR, 4.8; 95% CI, 1.2-18.6). CONCLUSION: In this study, the presence of a complicated diagnosis and a number of other factors were associated with an SSI after pediatric intracranial surgery for craniofacial malformations. We speculate that a complicated diagnosis may be a marker for other factors that contribute to the risk of an SSI. Future studies investigating SSIs after intracranial surgery should consider these factors. PMID- 15792513 TI - Phase II clinical trial of moderate hypothermia after severe traumatic brain injury in children. AB - OBJECTIVE: To determine whether moderate hypothermia (HYPO) (32-33 degrees C) begun in the early period after severe traumatic brain injury (TBI) and maintained for 48 hours is safe compared with normothermia (NORM) (36.5-37.5 degrees C). METHODS: After severe (Glasgow Coma Scale score < or =8) nonpenetrating TBI, 48 children less than 13 years of age admitted within 6 hours of injury were randomized after stratification by age to moderate HYPO (32-33 degrees C) treatment in conjunction with standardized head injury management versus NORM in a multicenter trial. An additional 27 patients were entered into a parallel single-institution trial of excluded patients because of late transfer or consent (delayed in transfer >6 h but within 24 h of admission), unknown time of injury (e.g., child abuse), and adolescence (e.g., aged 13-18 yr). Assessments of safety included mortality, infection, coagulopathy, arrhythmias, and hemorrhage as well as ability to maintain target temperature, mean intracranial pressure (ICP), and percent time of ICP less than 20 mm Hg during the cooling and subsequent rewarming phases. Additionally, assessments of neurocognitive outcomes were obtained at 3 and 6 months of follow-up. RESULTS: Moderate HYPO after severe TBI in children was found to be safe relative to standard management and NORM in children of all ages and in children with delay of initiation of treatment up to 24 hours. Although there was decreased mortality in HYPO in both studies, there was an increased potential for arrhythmias with HYPO, although they were manageable with fluid administration or rewarming. Additionally, there was a reduction in mean ICP during the first 72 hours after injury in both studies, although rebound ICP elevations in HYPO compared with those in NORM were noted for up to 10 to 12 hours after rewarming. Although functional outcome at 3 or 6 months did not differ between treatment groups, functional outcome tended to improve from the 3- to 6-month cognitive assessment in HYPO compared with NORM, although the sample size was too small for any definitive conclusions. CONCLUSION: HYPO is likely a safe therapeutic intervention for children after severe TBI up to 24 hours after injury. Further studies are necessary and warranted to determine its effect on functional outcome and intracranial hypertension. PMID- 15792514 TI - Assessment of the learning curve for lumbar microendoscopic discectomy. AB - OBJECTIVE: An understanding of the learning curve of a new surgical procedure is essential for its safe clinical integration, teaching, and assessment. This knowledge is currently deficient for lumbar microendoscopic discectomy (MED). The present article aims to profile the learning curve for MED of an individual surgeon in a hospital not previously exposed to this procedure. METHODS: The first 35 cases of MED for posterolateral lumbar disc prolapse causing radiculopathy performed at the Princess Alexandra Hospital, Brisbane, Australia, were studied prospectively. The learning curve was assessed using surgery time, conversion rate, complication rate, surgeon "comfort," and key learning steps. RESULTS: The duration of surgical operating time decreased over the course of the study, initially rapidly and then more gradually. There were three conversions to open discectomy in the first 7 cases and none in the next 28 cases. The complexity of cases increased over the series, and the complication rate decreased. The asymptote of the learning curve seems to be approximately 30 cases. The specific learning tasks of MED include lateral lamina radiology, scope vision, visuospatial orientation, smaller field of view, angle of approach and tube position, and care and handling of endoscope equipment. CONCLUSION: A learning curve for MED has been demonstrated. Further assessment of this curve for a population of surgeons is necessary before a clinical assessment of open discectomy versus MED can be embarked upon. PMID- 15792515 TI - Monitoring the expression profiles of integrins and adhesion/growth-regulatory galectins in adamantinomatous craniopharyngiomas: their ability to regulate tumor adhesiveness to surrounding tissue and their contribution to prognosis. AB - OBJECTIVE: The purpose of this study was to identify biological markers that may be involved in the adhesiveness of craniopharyngiomas to optical chiasms and/or pituitary stalks. METHODS: We determined the complete pattern of integrin expression in three craniopharyngiomas by means of a complementary deoxyribonucleic acid microarray. We quantitatively determined the levels of immunohistochemical expression of the different integrins in a series of 37 cases and the pattern of immunohistochemical expression of 10 extracellular matrix components (acting as integrin ligands) in 7 optical chiasms and 11 pituitary stalks. We also quantitatively (computer-assisted microscopy) determined the levels of immunohistochemical expression of galectin-1, -3, -4, -7, and -8 in 50 adamantinomatous craniopharyngiomas. RESULTS: The present study shows that at both the ribonucleic acid and protein levels, adamantinomatous craniopharyngiomas express the alpha2, alpha6, alpha(v), beta1, beta5, and beta8 integrin subunits, whereas optical chiasms and pituitary stalks express vitronectin, thrombospondin, and various forms of collagens. CONCLUSION: Our data suggest that at least part of the adhesiveness of craniopharyngiomas to the surrounding tissue, such as optical chiasms and pituitary stalks, could be explained by the interactions between alpha(2beta1) integrin expressed by craniopharyngiomas and collagens on the one hand, and vitronectin expressed by the surrounding tissue on the other hand. In addition, a Cox regression analysis has revealed that the levels of galectin-4 contribute significant information toward the delay in recurrence independently of surgical status. PMID- 15792516 TI - Radiosurgery for cranial base chordomas and chondrosarcomas. AB - OBJECTIVE: To evaluate the efficacy and toxicity of radiosurgery in the treatment of cranial base chordoma and chondrosarcoma. METHODS: We reviewed 29 patients with cranial base chordoma (n = 25) or chondrosarcoma (n = 4) who underwent stereotactic radiosurgery between September 1990 and December 2002. The median patient age was 45 years (range, 10-81 yr). Nineteen patients also had radiation therapy before or in conjunction with radiosurgery (median dose, 50.4 Gy). The median tumor volume was 14.4 cm3 (range, 0.6-65.1 cm3). The median tumor margin dose was 15 Gy (range, 10-20 Gy); the median maximum radiation dose was 30 Gy (range, 20-40 Gy). Median clinical and imaging follow-up periods were 4.8 and 4.5 years, respectively. RESULTS: Seven chordoma patients (28%) had tumor progression (in-field, n = 3; out-of-field, n = 4), whereas 18 had stable disease or tumor shrinkage. No patient with a chondroid chordoma had tumor enlargement. The actuarial tumor control rates were 89 and 32% at 2 and 5 years, respectively. All 4 patients with chondrosarcoma had tumor control. Clinically, 7 patients (24%) had improvement of pretreatment symptoms, 16 (55%) remained stable, and 6 (21%) worsened. Three patients with tumor progression died. Ten patients (34%) had radiation-related complications. Complications included cranial nerve deficits (n = 6), radiation necrosis (n = 5), and pituitary dysfunction (n = 3). Patients having radiosurgery alone had no toxicity. CONCLUSION: Cranial base chordomas and chondrosarcomas remain a formidable management challenge. Radiosurgery as an adjunct to surgical resection provides in-field tumor control for some patients, but radiation-related complications are relatively high, especially when radiosurgery is combined with fractionated radiation therapy. PMID- 15792517 TI - Evaluation of the stability of small ruptured aneurysms with a small neck after embolization with Guglielmi detachable coils: correlation between coil packing ratio and coil compaction. AB - OBJECTIVE: Because it is difficult to predict the compaction of Guglielmi detachable coils (GDCs) after endovascular surgery for aneurysms, we studied the relationship between the coil packing ratio and compaction. Here, we propose a simple method for the preoperative estimation of coil compaction. Using follow-up angiograms, we studied the timing and degree of coil compaction in small terminal and side-wall aneurysms with narrow necks. METHODS: We studied 62 patients with acute ruptured intracranial aneurysms that were small (<10 mm), had a small neck (<4 mm), and were coil embolized with GDC-10s. The aneurysmal volume was calculated using the equation V = 4/3pi(a/2) x (b/2) x (c/2), where a, b, and c are the aneurysmal height, length, and width in millimeters, respectively. The coil volume was calculated using the equation V = pi(p/2)2 x l x 10, where p represents the GDC-10 coil diameter (0.25 mm) and l is the coil length. We recorded the maximum prospective coil length, L, as that corresponding with the volume of packed coils occupying 30% of the aneurysmal volume. Therefore, L was calculated as L (cm) = 0.3 x a x b x c, and the coil packing ratio was expressed as packed coil length/L x 100. Angiographic follow-up studies were generally performed at 3 months and 1 and 2 years after endovascular surgery. We considered coil compaction exceeding 2 mm as major compaction and recorded minor compaction when it was less than 2 mm of the empty reappeared space in the embolized aneurysm. Aneurysmal location was recorded as terminal or side wall. RESULTS: Of the 62 patients, 16 (25.8%) manifested angiographic coil compaction (10 minor and 6 major compactions); the mean coil packing ratio was 51.9 +/- 13.4%. The mean coil packing ratio in the other 46 patients was 80.5 +/- 20.2%, and the difference was statistically significant (P < 0.01). In all 6 patients with major compaction, the mean packing ratio was less than 50% and all underwent re embolization after a mean of 24.9 +/- 1.1 months. The 10 patients with minor compaction were conservatively treated, and the degree of compaction did not change during a mean period of 24 months. We detected 93.8% of the compactions within 12 months of coil placement. The aneurysm was of the terminal type in 5 of the 6 patients with major coil compaction. CONCLUSION: In patients who underwent embolization with GDC-10s of aneurysms that were small and had a small neck, the optimal coil packing ratio could be identified with the formula 0.3 x a x b x c. The probability of coil compaction was significantly higher when the coil packing ratio was less than 50%. To detect coil compaction after embolization, follow-up angiograms must be examined regularly for at least 12 months. To detect major coil compaction in patients with terminal type aneurysms, angiographic follow-up should not be shorter than 24 months. PMID- 15792518 TI - Calcium alginate gel as a biocompatible material for endovascular arteriovenous malformation embolization: six-month results in an animal model. AB - OBJECTIVE: We sought to expand our assessment of calcium alginate as an embolic agent in an animal model of a cerebral arteriovenous malformation (AVM). The objective of this study was to assess the long-term biocompatibility and stability of calcium alginate in AVM swine models that survived from 1 to 6 months. METHODS: The swine model included a carotid-jugular anastomosis to redirect flow to the rete mirabile (RM), thereby simulating flow to an AVM. Alginate and the reactive component, calcium chloride, were injected from double lumen or concentric-tube microcatheters to form an occlusion of the RM feeding vessel and the inferior portion of the RM. RESULTS: Angiography and histology verified complete occlusion of the RM feeding vessel for up to 6 months in eight of nine swine. Blood flow remained open to the superior portion of the RM and the circle of Willis. No evidence of downstream calcium alginate gel was seen in the follow-up angiograms or the histological preparations of the circle of Willis. A minor bioactive response to the alginate gel was noted at 1 month, yet no degenerative or inflammatory response was seen. At 6 months, there was moderate fibrous tissue around the alginate, which further sealed off flow to the embolized areas of the RM. CONCLUSION: Over a period of 6 months, calcium alginate was an effective endovascular occlusion material that blocked blood flow to the inferior portion of the RM. The chronic AVM model verified the long-term stability and biocompatibility of calcium alginate. PMID- 15792519 TI - Expression of telomeric repeat binding factor-1 in astroglial brain tumors. AB - OBJECTIVE: In human somatic cells, telomeres shorten with successive cell divisions, resulting in progressive genomic instability, altered gene expression, and cell death. Recently, telomere-specific deoxyribonucleic acid-binding proteins, such as telomeric repeat binding factor-1 (TRF1), have been proposed as candidates for the role of molecules regulating telomerase activity, and they have been suggested to play key roles in the maintenance of telomere function. The present study was designed to assess TRF1 expression in human astroglial brain tumors and to speculate on the clinical implications of its expression. METHODS: Twenty flash-frozen surgical specimens obtained from adult patients who underwent craniotomy for microsurgical tumor resection, histologically verified as World Health Organization Grade II to IV astrocytomas, were used. Expression of TRF1 in astrocytomas of different grades was studied by means of both immunohistochemical and Western blotting analysis. The correlation between the extent of TRF1 expression and histological grading, performance status, and length of survival of patients underwent statistical analyses. RESULTS: TRF1 was expressed in all tumor samples. The level of its expression was variable, decreasing from low-grade through high-grade astrocytomas (P = 0.0032). TRF1 expression correlated with the patient's length of survival (P < 0.001) and performance status (P < 0.001) and proved to be an independent indicator of length of survival. CONCLUSION: Our findings suggest that the loss of TRF1 expression capability, as a result of down-regulation of TRF1 expression in malignant gliomas cells, may play a role in the malignant progression of astroglial brain tumors. PMID- 15792520 TI - The cerebellum modulates rodent cortical motor output after repetitive somatosensory stimulation. AB - OBJECTIVE: To analyze the possible role of the cerebellum in the modulation of cortical motor output associated with repetitive electrical stimulation of the sciatic nerve in the rat. METHODS: A sustained somatosensory stimulation induces an increase in the intensity of the response of the rodent motor cortex. Wistar rats were anesthetized for surgical preparation using a continuous infusion of chloral hydrate. We analyzed the response evoked by electrical stimulation of the right motor cortex before (basal condition) and after peripheral electrical stimulation of the left sciatic nerve in rats with no cerebellar intervention (n = 6), and in control rats with Ringer's infusion via a microdialysis probe (n = 8) implanted in the left cerebellar nuclei. In addition, we investigated the effects of 1) the administration of ethanol (20 mmol/L) in the left cerebellar nuclei (n = 5); 2) the administration of tetrodotoxin (10 micromol/L), a sodium channel blocker, in the left cerebellar nuclei (n = 5); 3) electrical stimulation by deep cerebellar stimulation (frequency 100 Hz) on the left side (n = 5); or 4) electrical stimulation of the cerebellar nuclei on the right side (100 Hz; n = 6). For peripheral stimulation, all of the animals received 1 hour of electrical stimulation. Trains of stimulation consisted of five stimuli (duration of 1 stimulus, 1 ms) at a rate of 10 Hz. During stimulation of the motor cortex, peak to-peak amplitudes in responses of the left calf muscle were analyzed. Motor threshold was defined as the lowest intensity eliciting at least 5 of 10 evoked responses with an amplitude greater than 20 muV. The intensity used was 130% of the motor threshold. RESULTS: In the basal condition (before repetitive stimulation), amplitudes of motor responses were similar in the six groups of rats (P = 0.40). In rats without cerebellar intervention, peripheral electrical stimulation was associated with an increase of motor response to 147.4 +/- 8.5% of baseline (P < 0.001). In rats with Ringer's infusion, the motor response increased to 141.6 +/- 7.9% of baseline (P < 0.001). The administration of ethanol in the cerebellum prevented the enhancement of the response ipsilaterally. The mean +/- standard deviation (SD) of motor responses was 105.7 +/- 6.2% of baseline measurements after stimulation of the sciatic nerve (P = 0.36). The same observation was made after the infusion of tetrodotoxin (mean +/- SD of motor responses: 107.1 +/- 7.4% after peripheral stimulation [P = 0.19] and after electrical stimulation of the cerebellum on the left side [mean +/- SD of motor responses, 104.3 +/- 8.5% after peripheral stimulation, P = 0.40]). However, electrical stimulation of cerebellar nuclei on the right side did not impair the modulation of cortical motor output by sciatic nerve stimulation (mean +/- SD of motor responses, 148.4 +/- 5.8% after peripheral stimulation, P < 0.001). CONCLUSION: Until now, the increase of motor output after peripheral nerve stimulation has been considered as a plasticity directly and solely dependent on cortical structures. We demonstrate that the cerebellum plays a key role in this form of neural plasticity. PMID- 15792521 TI - Erythropoietin and erythropoietin receptor expression after experimental spinal cord injury encourages therapy by exogenous erythropoietin. AB - OBJECTIVE: Erythropoietin (EPO) is a pleiotropic cytokine originally identified for its role in erythropoiesis. Recent studies have demonstrated that EPO and its receptor (EPO-R) are expressed in the central nervous system, where EPO exerts neuroprotective functions. Because the expression of the EPO and EPO-R network is poorly investigated in the central nervous system, the aim of the present study was to investigate whether the resident EPO and EPO-R network is activated in the injured nervous system. METHODS: A well-standardized model of compressive spinal cord injury in rats was used. EPO and EPO-R expression was determined by immunohistochemical analysis at 8 hours and at 2, 8, and 14 days in the spinal cord of injured and noninjured rats. RESULTS: In noninjured spinal cord, weak immunohistochemical expression of EPO and EPO-R was observed in neuronal and glial cells as well as in endothelial and ependymal cells. In injured rats, a marked increase of expression of EPO and EPO-R was observed in neurons, vascular endothelium, and glial cells at 8 hours after injury, peaking at 8 days, after which it gradually decreased. Two weeks after injury, EPO immunoreactivity was scarcely detected in neurons, whereas glial cells and vascular endothelium expressed strong EPO-R immunoreactivity. CONCLUSION: These observations suggest that the local EPO and EPO-R system is markedly engaged in the early stages after nervous tissue injury. The reduction in EPO immunoexpression and the increase in EPO-R staining strongly support the possible usefulness of a therapeutic approach based on exogenous EPO administration. PMID- 15792523 TI - Clinicoradiological review: symptomatic superior cerebellar artery aneurysm imitating cerebellopontine angle tumor. PMID- 15792522 TI - Effects of antithrombin III on myeloperoxidase activity, superoxide dismutase activity, and malondialdehyde levels and histopathological findings after spinal cord injury in the rat. AB - OBJECTIVE: Among the many possible mechanisms of the secondary spinal cord injury (SCI), microcirculatory disturbances as a result of activated leukocyte-induced endothelial cell injury is important because it is potentially treatable and reversible. Currently, clinically available pharmacological agents for treatment of acute SCI do not inhibit neutrophil activation. The effect of antithrombin III (AT-III) on neutrophil activation was studied in rats with SCI produced with an aneurysm clip on the T2-T7 segments. METHODS: Forty rats were randomly allocated to four groups. Group I (10 rats) was killed to provide normal spinal cord tissue for testing. Group II (10 rats) underwent a six-segment laminectomy for the effects of total laminectomy to be determined. In Group III, 10 rats underwent a six-segment laminectomy and SCI was produced by extradural compression of the exposed cord. The same procedures were performed in 10 rats in Group IV, but they also received one (250 IU/kg) intraperitoneal injection of AT-III immediately after the injury and a second dose 24 hours later. The animals from Groups II through IV were killed 48 hours after the trauma. The effect of AT-III on the myeloperoxidase activity, superoxide dismutase activity, and malondialdehyde levels and histopathological findings were studied. RESULTS: Myeloperoxidase activity, superoxide dismutase activity, and malondialdehyde levels were significantly lower and there was less histopathological damage in the AT-III treatment group than in the trauma group. CONCLUSION: The results demonstrate that AT-III treatment may reduce secondary structural changes in damaged rat spinal cord tissue by inhibiting leukocyte activation. PMID- 15792524 TI - Neurosurgical oncology at the university of Texas M. D. Anderson Cancer Center: its genesis and evolution. AB - The practice of neurosurgery at The University of Texas M. D. Anderson Cancer Center began in 1944 with one neurosurgeon among the 11 physicians present in a makeshift 16-room outpatient clinic at a temporary location. Neurosurgical oncology evolved as the hospital did, first as a neurosurgery service in 1951, then as the Section of Neurosurgery within the Department of Head and Neck Surgery in 1979, and finally, as the Department of Neurosurgery in 1990. Although M. D. Anderson is now one of the largest institutions in the world devoted exclusively to cancer patient care, research, education, and prevention, it has an unusual history, which is reviewed in terms of the institution's origin in 1941, its development under three presidents, and its fostering of neurosurgical oncology. We chronicle the growth and development of the department from 1990 to 2003 and describe the unique opportunities it presents for surgical innovation, for clinical and basic research, for training residents and fellows, and for multidisciplinary collaboration in neurosurgical oncology. PMID- 15792525 TI - Development of stereotactic neurosurgery in China. AB - As a brief reflection on the development of stereotactic neurosurgery, this article reviews its founding and early history in China. PMID- 15792526 TI - Herophilus of Chalcedon: a pioneer in neuroscience. AB - A review of the history of ancient medicine reveals that most of the knowledge is concentrated in the studies of a few scientists. The best-known names include Hippocrates, Rufus of Ephesus, Celsus, and Galen. The survival of their works throughout the ages has been the most important factor contributing to their popularity. However, there are other scientists who made great contributions to science, but whose writings have been lost or destroyed over the course of time. As a result, their names are not as well known as those of others and the value of their contributions is not appreciated. With the improvement of communication technology in the past 50 years, links between the studies of ancient science can be made more effectively and scientists who have remained hidden under the shade of time have begun, after thousands of years, to receive the appreciation they deserve. In the field of neuroscience, the historical record focuses on Galen of Pergamon. But, when his marvelous works are carefully studied, it is interesting to note two names he frequently referenced: Herophilus (335-280 BC) and Erasistratus (310-250 BC). These two scientists were the first to place scientific value on the dissection of the human body. Herophilus is considered the father of scientific anatomy, and Erasistratus was the first experimental physiologist. Attracted by the prospect of material advancement and eminent students, both migrated from their homes in Asia Minor to Alexandria. The works of Herophilus and Erasistratus have been lost entirely, but some details of their teachings may be recovered from the writings of Galen. In this study, we focus on Herophilus, a master of ancient medicine, whose important discoveries about the human body formed the basis for positive science and the foundation for neuroscience. PMID- 15792527 TI - Treatment of phenylketonuria-associated tremor with deep brain stimulation: case report. AB - OBJECTIVE AND IMPORTANCE: Phenylketonuria (PKU) is an inborn error of metabolism that causes severe neurological impairment, despite dietary treatment. We present a case of PKU-induced cerebellar tremor treated with deep brain stimulation. There have been no previously reported cases of a patient with a PKU tremor treated with deep brain stimulation. CLINICAL PRESENTATION: A 36-year-old male patient with PKU presented with signs of cerebellar disease including dysmetria, resting tremor, and intention tremor in the left upper extremity. INTERVENTION: A deep brain stimulation electrode was placed in the ventral intermediate nucleus of the right thalamus. CONCLUSION: Immediately after surgery, the patient had nearly complete resolution of intention tremor in the left arm. His resting tremor in the left hand was also greatly improved. The 30-month follow-up examination revealed maintenance of the immediate postoperative improvement. PMID- 15792528 TI - Intramedullary ectopic choroid plexus: report of a rare case. AB - OBJECTIVE AND IMPORTANCE: Intracranial cysts containing an ectopic choroid plexus or choroid plexus-like tissue have seldom been described in the literature. However, there has been no report of a spinal intramedullary cyst containing an ectopic choroid plexus. This is the first case report in the available literature of an ectopic choroid plexus tissue in the spinal cord. CLINICAL PRESENTATION: A 30-year-old man presented with complaints of progressive descending weakness of both upper limbs and increasing stiffness of the lower limbs along with numbness of all four limbs without a history of any bowel or bladder disturbances. Examination was suggestive of a C5-T2 intramedullary lesion. Magnetic resonance imaging revealed a C6-T2 intramedullary cystic lesion along with a small anterosuperiorly placed lesion enhancing with contrast. INTERVENTION: A C6-T2 laminotomy and exploration of the intramedullary cyst and gross total microsurgical excision of the reddish vascular frond-like structure resembling the choroid plexus were performed along with a syringostomy. A laminoplasty with miniplates and screws was performed. Histopathological and immunohistochemical studies revealed a normal choroid plexus. The patient has been followed for 1 year and has demonstrated symptomatic improvement. CONCLUSION: Although there have been rare case reports of drop metastasis of choroid plexus papillomas in the spine, this is the first case report in the available literature of normal but ectopic choroid plexus tissue in the spinal cord. PMID- 15792529 TI - Bilateral reversible deafness after surgery for unilateral epidermoid tumor: an unusual complication: case report. AB - OBJECTIVE AND IMPORTANCE: We present an unusual complication of bilateral hearing loss after surgery for unilateral epidermoid tumor situated at the right pontocerebellar junction. The neurosurgeon must know the possible causes of this catastrophic complication and should take every possible measure to avoid it. CLINICAL PRESENTATION: A 43-year-old male patient presented with progressive hearing loss in the right ear. A radiological examination demonstrated a mass lesion at the right pontocerebellar junction. A pure tone audiogram revealed sensorineural hearing impairment of the right ear. A brainstem auditory evoked potential test confirmed this with reduced amplitudes and a prolonged I-V interval. INTERVENTION: The patient was operated on via a right paramedian suboccipital craniectomy. The tumor was completely excised, and all of the cranial nerves were preserved during the operation. Postoperatively, the patient developed total bilateral hearing loss, which improved significantly over time. CONCLUSION: Controlled cerebrospinal fluid drainage during exposure and tumor excision may reduce the possibility of shift and traction on the neural structures and help to prevent ischemic injury. Frequent irrigation of the surgical field and minimizing the spilling of the tumor contents may reduce the risk of chemical irritation. The use of corticosteroids may also help during the postoperative period. PMID- 15792530 TI - Hemothorax as a complication after anterior cervical discectomy: case report. AB - OBJECTIVE AND IMPORTANCE: Anterior cervical discectomy is one of the most frequently performed surgical procedures of the cervical spine. CLINICAL PRESENTATION: We describe a patient with a postoperative hemothorax as a complication after an anterior cervical discectomy. INTERVENTION: The patient was intubated, and reoperation and bronchoscopy were performed. CONCLUSION: To the best of our knowledge, such a complication has not been described before in the English-language literature. We conclude that hemothorax is a rare, although serious, complication after anterior cervical discectomy and should be included in the differential diagnosis for patients with respiratory failure after an anterior cervical discectomy. PMID- 15792531 TI - Decompressive hemicraniectomy for acute disseminated encephalomyelitis: case report. AB - OBJECTIVE AND IMPORTANCE: Acute disseminated encephalomyelitis (ADEM) varies widely in symptoms and severity. Some cases are associated with massive life threatening cerebral edema refractory to conventional medical management. CLINICAL PRESENTATION: A 51-year-old woman with ADEM who developed severe brain swelling and herniation despite aggressive medical management is described. INTERVENTION: A decompressive hemicraniectomy and durotomy led to rapid improvement and an excellent outcome. CONCLUSION: This case report reinforces the place of this procedure in the armamentarium of treatment options for patients with medically refractory brain swelling and elevated intracranial pressure caused by ADEM. The potential for an increase in the incidence of ADEM with more frequent smallpox vaccinations emphasizes the significance of redefining the full range of management options for this treatable disease. PMID- 15792532 TI - The mouse chemerin receptor gene, mcmklr1, utilizes alternative promoters for transcription and is regulated by all-trans retinoic acid. AB - CMKLR1 (chemoattractant-like receptor 1) is a G-protein-coupled receptor implicated in cartilage and bone development and is expressed in organs like the parathyroid gland, brain, and lung. The receptor is also expressed in dendritic cells and in macrophages where it acts as a co-receptor for entry of HIV/SIV isolates into human CD4(+) cells. Recently, a protein named "chemerin" (also known as TIG2) was isolated from human inflammatory fluids and hemofiltrate and found to be the endogenous ligand for CMKLR1. We have previously described the genomic organization of the cmklr1 gene and characterized its promoter in mouse neuroblastoma NB4 1A3 cells. In the present study we identify a second transcript, cmklr1b, in mouse microglia BV2 cells. Cmklr1b is transcribed from an alternative promoter with a transcription start site located 6780 bp downstream of the previously identified exon 1 (cmklr1a). The cmklr1b promoter lacks a TATA box but contains two CCAAT boxes in opposite directions. 5' Deletion analysis of the promoter region in BV2 cells using a luciferase reporter gene assay indicates two regions, between 623-755 bp and 56-125 bp upstream of transcription start site, to be important for promoter function. The proximal promoter region includes both CCAAT boxes, and site-directed mutagenesis separately within these elements revealed that only the forward CCAAT element was important for transcription. Although the forward CCAAT element is essential for transcription electrophoretic mobility shift and super-shift assays demonstrated that both CCAAT elements actually bind nuclear proteins from BV2 cells and identified the binding factor as NFY. Real-time reverse transcriptase-PCR experiments of cmklr1b expression in all-trans retinoic acid (ATRA)- stimulated BV2 cells showed strong up-regulation of receptor transcript. Luciferase reporter gene assay of the promoter in ATRA-stimulated BV2 cells confirmed that transcriptional activity of the cmklr1b promoter is increased by ATRA. However, deletion analysis could not identify an ATRA-responsive element within the promoter region suggesting that gene activation is likely to occur through alternative mechanisms. The results emphasise a possible role of cmklr1 in bone modelling. PMID- 15792533 TI - Identification of three duplicated Spin genes in medaka (Oryzias latipes). AB - Gene and genomic duplications are very important and frequent events in fish evolution, and the divergence of duplicated genes in sequences and functions is a focus of research on gene evolution. Here, we report the identification and characterization of three duplicated Spindlin (Spin) genes from medaka (Oryzias latipes): OlSpinA, OlSpinB, and OlSpinC. Molecular cloning, genomic DNA Blast analysis and phylogenetic relationship analysis demonstrated that the three duplicated OlSpin genes should belong to gene duplication. Furthermore, Western blot analysis revealed significant expression differences of the three OlSpins among different tissues and during embryogenesis in medaka, and suggested that sequence and functional divergence might have occurred in evolution among them. PMID- 15792534 TI - Genotyping of hepatitis B virus in Malaysia based on the nucleotide sequence of preS and S genes. AB - Hepatitis B virus (HBV) has been classified into eight genotypes, designated A-H. These genotypes are known to have distinct geographic distributions. The clinical importance of genotype-related differences in the pathogenicity of HBV has been revealed recently. In Malaysia, the current distribution of HBV remains unclear. The aim of this study was to determine the genotypes and subtypes of HBV by using PCR, followed by DNA sequencing, as well as to analyse the mutations in the immunodominant region of preS and S proteins. The S gene sequence was determined from HBV DNA of four apparently healthy blood donors' sera and three sera from asymptomatic chronic hepatitis B carriers. Of this batch of sera, the preS gene sequence was obtained from HBV DNA from three out of the four blood donors and two out of the three chronic carriers. Due to insufficient sera, we had to resort to using sera from another blood donor to make up for the sixth DNA sequence of the preS gene. Based on the comparative analysis of the preS sequences with the reported sequences in the GenBank database, HBV DNA from two normal carriers was classified as genotype C. Genotype B was assigned to HBV from one blood donor and two hepatitis B chronic carriers, whereas HBV of one chronic carrier was of genotype D. Based on the S gene sequences, HBV from three blood donors was of genotype C, that of one blood donor and one chronic carrier was of genotype B, and the remaining, of genotype D. In the five cases where both preS and S gene sequences were determined, the genotypes assigned based on either the preS or S gene sequences were in concordance. The nature of the deduced amino acid (aa) sequences at positions 125, 127, 134, 143, 159, 161 and 168 of the S gene enabled the classification of these sequences into subtypes, namely, adrq+, adw2 and ayw2. The clustering of our DNA sequences into genotype groups corresponded to their respective subtype, that is, adw2 in genotype B, adrq in genotype C and ayw in genotype D. Analysis of the point mutations revealed that five of the sequences contained aa substitutions at immunodominant epitopes involved in B or/and T cell recognition. In conclusion, despite the low numbers of samples studied, due to budget constraints, these data are still worthwhile reporting, as it is important for the control of HBV infections. In addition, the genotype and mutational data obtained in this study may be useful for designing new treatment regimes for HBV patients. PMID- 15792535 TI - Systematic variation of interfacial phase reactivity in dental nanocomposites. AB - This study was designed to determine the effect of varying the chemistry of the interfacial phase on critical composite properties in dental nanocomposite materials. Silica nanoparticles were silanized with varying ratios of 3 methacryloxypropyltrimethoxysilane (MPTMS) and octyltrimethoxysilane (OTMS) while keeping the total amount of silane constant at 10% by mass fraction relative to the mass of filler. The silanized nanoparticles were mixed into a dimethacrylate resin (60% filler by mass fraction). The mechanical properties of the uncured pastes were assessed by compression testing between parallel plates. The composites were photo-cured and tested by biaxial flexure and three-point bend flexure testing. Fracture surfaces were analyzed by field-emission scanning electron microscopy (FE-SEM). At maximized filler mass fractions, the workabilities of the uncured pastes were better maintained as the fraction of OTMS in the interphase increased relative to MPTMS. The flexure strengths and moduli of the MPTMS silanized and dual silanized composites were similar but decreased as OTMS mass fractions in the silane mixture increased to 7.5% and 10%. FE-SEM images revealed evidence for phase separation in the composites containing silica silanized with high fractions of OTMS. Among the potential practical benefits of dual silanized nanoparticles are the improved workability of composite pastes with higher filler loadings that should lead to higher modulus composites with lower polymerization shrinkage. PMID- 15792536 TI - Evidence of chemical bonding to hydroxyapatite by phosphoric acid esters. AB - Phosphoric acid esters (PAEs) have been used as a self-etching primer for composite-to-enamel bonding in adhesive dentistry. However, the chemical mechanism of their interactions with hydroxyapatite (HA) is not clear. In the present study, HA particles were mixed with Resulcin AquaPrime (Merz Co.) priming agent that contains a mixture of PAEs, and dried. The primer, HA and the mixture of both were analyzed by FTIR. Primed and untreated HA discs were analyzed with attenuated total reflectance (ATR). After AquaPrime+D(2)O (1:1) was mixed with HA or Ca(OD)(2) for 48 h, the primer and both mixtures were analyzed with (31)P NMR in D(2)O. The solid and the liquid separated from both mixtures were analyzed with (31)P NMR in CDCl(3). The primer's characteristic bands (nu(CO), nu(CC)) were found on the primer-subtracted mixture's spectrum and the primed HA disc. (31)P NMR data indicated that the reactions of PAEs with HA produced PAEs-HA complexes, and were not a simple acid-base reaction like those with Ca(OD)(2), either in liquid or in solid. It is concluded that phosphoric acid esters can decalcify and adhere to hydroxyapatite simultaneously. PMID- 15792537 TI - Surface characterization of silver-doped bioactive glass. AB - A bioactive glass belonging to the system SiO(2)-CaO-Na(2)O was doped with silver ions by ion exchange in molten salts as well as in aqueous solution. The ion exchange in the solution was done to check if it is possible to prepare an antimicrobial material using a low silver content. The doped glass was characterized by means of X-ray diffraction, SEM observation, EDS analysis, bioactivity test (soaking in a simulated body fluid), leaching test (GFAAS analyses) and cytotoxicity test. It is demonstrated that these surface silver doped glasses maintain, or even improve, the bioactivity of the starting glass. The measured quantity of released silver into simulated body fluid compares those reported in literature for the antibacterial activity and the non-cytotoxic effect of silver. Cytotoxicity tests were carried out to understand the effect of the doped surfaces on osteogenic cell adhesion and proliferation. PMID- 15792538 TI - Synthesis of a poly(L-lysine)-calcium phosphate hybrid on titanium surfaces for enhanced bioactivity. AB - Titanium has been a successful implant material owing to its excellent strength to weight ratio, toughness, and bioinert oxide surface. Significant progress has been made on the improvement of titanium's bioactivity by coating its oxide surface with calcium phosphates and bioactive molecules. Here, we report on the coating of titanium with a poly(L-lysine)-calcium phosphate hybrid material with a nanoscale texture. This hybrid coating was grown by first nucleating seed crystals of calcium phosphate, directly on the Ti surface and then exposing this surface to solutions containing Ca(2+), PO(4)(3-), and poly(L-lysine). The resultant hybrid coating was characterized by electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray photoelectron spectroscopy, and elemental analysis. This material contained 14% by weight poly(L-lysine), and this organic component decreased greatly the dimensions of the surface features, thus enhancing surface area relative to the inorganic control. The highly textured hybrid material was more susceptible than the control to acidic and enzymatic degradation. The amino acid cysteine was covalently linked to the hybrid material, demonstrating the potential of this coating for further functionalization. These hybrid coatings may prove useful in enhancing the bioactivity of titanium. PMID- 15792539 TI - TEGDMA induces mitochondrial damage and oxidative stress in human gingival fibroblasts. AB - Free monomers including triethylene glycol dimethacrylate (TEGDMA) are released by resin composite. Recent studies in vitro have demonstrated that TEGDMA induced GSH depletion and production of radical oxygen species (ROS) in human gingival fibroblasts (HGF) but the exact mechanism of these events remains unclear. Our purpose is to investigate the origin of ROS production. TEGDMA induces a rapid (within 30 min) and drastic depletion of ATP concomitant with the GSH depletion. After 3h incubation, TEGDMA induced an increase of lipid peroxidation associated with LDH leakage. Our data also showed that TEGDMA produced damage at mitochondrial level. This is demonstrated by the collapse of mitochondrial membrane potential (MMP) in HGF treated with TEGDMA. The protective effect of carbonylcyanide m-chlorophenylhydrazone (CCCP), an uncoupler of oxidative phosphorylation on lipid peroxidation and LDH leakage suggests that mitochondria can be implicated in these events. Trolox, a soluble derivative of Tocopherol, weakly prevents ATP but not GSH depletion and totally protects the cells against lipid peroxidation, MMP collapse and cell death. Thus, the present results suggest that TEGDMA induces lipid peroxidation and mitochondrial damage, which contribute to cell death. PMID- 15792540 TI - The response of normal human osteoblasts to anionic polysaccharide polyelectrolyte complexes. AB - Polyelectrolyte complexes (PEC) were prepared from chitosan as the polycation and several synthesized functional anion polysaccharides, and their effects on cell attachment, morphology, proliferation and differentiation were estimated using normal human osteoblasts (NHOst). After a 1-week incubation, PEC made from polysaccharides having carboxyl groups as polyanions showed low viability of NHOst on it although the NHOst on it showed an enhancement in their differentiation level. On the other hand, NHOst on PEC made from sulfated or phosphated polysaccharides showed similar attachment and morphology to those on the collagen-coated dish. When the number of NHOst was estimated after 1 week, the number on the PEC was ranged from 70% to 130% of those on the collagen-coated dish, indicating few effects of these PEC on cell proliferation. In addition, NHOst on PEC films made from sulfated polysaccharides differentiated to a level very similar to that observed on the collagen-coated dish, indicating that these PEC films maintain the normal potential of NHOst to both proliferate and differentiate. Measurement of gap junctional intercellular communication of NHOst on PEC revealed that PEC did not inhibit communication, suggesting that PEC films have few effects on cell homeostasis. Thus, PEC made from the sulfated polysaccharide may be a useful material as a new scaffold for bone regeneration. PMID- 15792541 TI - Repair of an intercalated long bone defect with a synthetic biodegradable bone inducing implant. AB - Recombinant human bone morphogenetic protein (rhBMP)-2 in a block copolymer composed of poly-D,L-lactic acid with randomly inserted p-dioxanone and polyethylene glycol (PLA-DX-PEG) as a carrier and porous beta-tricalcium phosphate (beta-TCP) blocks were used to generate a new fully absorbable osteogenic biomaterial. The bone regenerability of the rhBMP-2/PLA-DX-PEG/beta TCP composite was studied in a critical-sized rabbit bone defect model. In an initial study, a composite of PLA-DX-PEG (250 mg) and beta-TCP (300 mg) loaded with or without rhBMP2 (50 microg) was implanted into a 1.5 cm intercalated bone defect created in a rabbit femur. Defects were assessed by biweekly radiography until 8 weeks postoperatively. The bony union of the defect was recognized only in the BMP-loaded group. To obtain further data on biomechanical and remodeling properties, another BMP-loaded composites group was made and observed up to 24 weeks. All defects were completely repaired without residual traces of implants. Anatomical and mechanical properties of the repaired bone examined by histology, 3-dimensional CT (3D-CT) and mechanical testing were essentially equivalent to the nonoperated-on femur at 24 weeks. These experimental results indicate that fully absorbable rhBMP-2/PLA-DX-PEG/beta-TCP is a promising composite having osteogenicity efficient enough for repairing large bone defects. PMID- 15792542 TI - The effect of the surface modification of titanium using a recombinant fragment of fibronectin and vitronectin on cell behavior. AB - The surface of titanium implants is in direct contact with host tissue and plays a critical role in determining biocompatibility. Fibronectin (FN) and vitronectin (VN) are major cell adhesive proteins found in the extracellular matrix (ECM) of various tissues, and in circulating blood. The aim of this study was to evaluate the engineered biomimetic surface of titanium by using recombinant fragment of FN(8-10) and VN(NTD) that contains the binding site for integrins. MC3T3-E1 cells seeded upon the FN(8-10)-coated titanium showed a marked increase in cell adhesion, proliferation, and differentiation over VN(NTD)-coated titanium. In addition, we confirmed that the surface properties of titanium prefer for FN(8 10) over VN(NTD) (p<0.05) in protein adhesion. These results suggest that the FN(8-10)-modified titanium surface can be used to improve the osseointegration of titanium implants by enhancing bone formation. PMID- 15792544 TI - Influence of insulin immobilization to thermoresponsive culture surfaces on cell proliferation and thermally induced cell detachment. AB - Temperature-responsive culture dishes immobilized with insulin have been fabricated and studied to shorten cell culture periods by facilitating more rapid cell proliferation. Cells are recovered as contiguous cell sheets simply by temperature changes. Functionalized culture dishes were prepared by previously reported electron beam grafting copolymerization of N-isopropylacrylamide (IPAAm) with its carboxylate-derivatized analog, 2-carboxyisopropylacrylamide (CIPAAm), having similar molecular structure to IPAAm but with carboxylate side chains to tissue culture polystyrene dishes. Insulin was then immobilized onto culture dishes through standard amide bond formation with CIPAAm carboxylate groups. Adhesion and proliferation of bovine carotid artery endothelial cells (ECs) were examined on these insulin-immobilized dishes. Insulin immobilization was shown to promote cell proliferation in serum-supplemented medium. Increasing the grafted CIPAAm content on the tissue culture surfaces reduces cell adhesion and proliferation, even though these surfaces contained increased amounts of immobilized insulin. This result implies that a discrete balance exists between the amount of CIPAAm-free carboxylate groups and immobilized insulin for optimum cell proliferative stimulation. Cells grown on the insulin-immobilized surfaces can be recovered as contiguous cell monolayers simply by lowering culture temperature, without need for exogenous enzyme or calcium chelator additions. In conclusion, insulin-modified thermoresponsive culture dishes may prove useful for advanced cell culture and tissue engineering applications since they facilitate cell proliferation, and cultured cells can be recovered as viable contiguous monolayers by merely reducing culture temperature. PMID- 15792543 TI - Multilineage differentiation of human mesenchymal stem cells in a three dimensional nanofibrous scaffold. AB - Functional engineering of musculoskeletal tissues generally involves the use of differentiated or progenitor cells seeded with specific growth factors in biomaterial scaffolds. Ideally, the scaffold should be a functional and structural biomimetic of the native extracellular matrix and support multiple tissue morphogenesis. We have previously shown that electrospun, three dimensional nanofibrous scaffolds that morphologically resemble collagen fibrils are capable of promoting favorable biological responses from seeded cells, indicative of their potential application for tissue engineering. In this study, we tested a three-dimensional nanofibrous scaffold fabricated from poly(epsilon caprolactone) (PCL) for its ability to support and maintain multilineage differentiation of bone marrow-derived human mesenchymal stem cells (hMSCs) in vitro. hMSCs were seeded onto pre-fabricated nanofibrous scaffolds, and were induced to differentiate along adipogenic, chondrogenic, or osteogenic lineages by culturing in specific differentiation media. Histological and scanning electron microscopy observations, gene expression analysis, and immunohistochemical detection of lineage-specific marker molecules confirmed the formation of three-dimensional constructs containing cells differentiated into the specified cell types. These results suggest that the PCL-based nanofibrous scaffold is a promising candidate scaffold for cell-based, multiphasic tissue engineering. PMID- 15792545 TI - Cytocompatibility of self-assembled beta-hairpin peptide hydrogel surfaces. AB - MAX1 is a 20 amino acid peptide that undergoes triggered self-assembly to form a rigid hydrogel. When dissolved in aqueous solutions, this peptide exists in an ensemble of random coil conformations rendering it fully soluble. The addition of an exogenous stimulus results in peptide folding into beta-hairpin conformation. This folded structure undergoes rapid assembly into a highly crosslinked hydrogel network. DMEM cell culture media is one stimulus able to initiate folding and consequent self-assembly of MAX1. The cytocompatibility of this gel towards NIH 3T3 murine fibroblasts is demonstrated. Gels were shown to be non-toxic to the fibroblast cells. MAX1 hydrogels also foster the ability of the cells to attach to the hydrogel scaffold in the absence or presence of serum proteins. Additionally MAX1 hydrogels were able to support fibroblast proliferation to confluency with little effect on the rheological properties of the scaffold. MAX1 hydrogels meet the preliminary mechanical and cytocompatibiltiy requirements of a tissue engineering scaffold. PMID- 15792546 TI - Tissue engineered cartilage on collagen and PHBV matrices. AB - Cartilage engineering is a very novel approach to tissue repair through use of implants. Matrices of collagen containing calcium phosphate (CaP-Gelfix), and matrices of poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) were produced to create a cartilage via tissue engineering. The matrices were characterized by scanning electron microscopy (SEM) and electron diffraction spectroscopy (EDS). Porosity and void volume analysis were carried out to characterize the matrices. Chondrocytes were isolated from the proximal humerus of 22 week-old male, adult, local albino rabbits. For cell type characterization, Type II collagen was measured by Western Blot analysis. The foams were seeded with 1x10(6) chondrocytes and histological examinations were carried out to assess cell-matrix interaction. Macroscopic examination showed that PHBV (with or without chondrocytes) maintained its integrity for 21 days, while CaP-Gelfix was deformed and degraded within 15 days. Cell-containing and cell-free matrices were implanted into full thickness cartilage defects (4.5 mm in diameter and 4 mm in depth) at the patellar groove on the right and left knees of eight rabbits, respectively. In vivo results at 8 and 20 weeks with chondrocyte seeded PHBV matrices presented early cartilage formation resembling normal articular cartilage and revealed minimal foreign body reaction. In CaP-Gelfix matrices, fibrocartilage formation and bone invasion was noted in 20 weeks. Cells maintained their phenotype in both matrices. PHBV had better healing response than CaP-Gelfix. Both matrices were effective in cartilage regeneration. These matrices have great potential for use in the repair of joint cartilage defects. PMID- 15792547 TI - The enhancement of osteoblast growth and differentiation in vitro on a peptide hydrogel-polyHIPE polymer hybrid material. AB - The objective of this study was to investigate the effect of combining two biomaterials on osteoblast proliferation, differentiation and mineralised matrix formation in vitro. The first biomaterial has a well-defined architecture and is known as PolyHIPE polymer (PHP). The second biomaterial is a biologically inspired self-assembling peptide hydrogel (RAD16-I, also called PuraMatrix) that produces a nanoscale environment similar to native extracellular matrix (ECM). Our work investigates the effect of combining RAD16-I with two types of PHP (HA (Hydroxyapatite)-PHP and H (Hydrophobic)-PHP) and evaluates effects on osteoblast growth and differentiation. Results demonstrated successful incorporation of RAD16-I into both types of PHP. Osteoblasts were observed to form multicellular layers on the combined biomaterial surface and also within the scaffold. Dynamic cell seeding and culturing techniques were compared to static seeding methods and produced a more even distribution of cells throughout the constructs. Cells were found to penetrate the scaffold to a maximum depth of 3 mm after 35 days in culture. There was a significant increase in cell number in H-PHP constructs coated with RAD16-I compared to H-PHP alone. Our results show that RAD16-I enhances osteoblast differentiation and indicates that the incorporation of this peptide provides a more permissive environment for osteoblast growth. We have developed a microcellular polymer containing a nanoscale environment to enhance cell: biomaterial interactions and promote osteoblast growth in vitro. PMID- 15792548 TI - The effect on osteoblast function of colocalized RGD and PHSRN epitopes on PEG surfaces. AB - Poly(ethylene glycol) hydrogels were synthesized with pendant peptide functionalities to examine the influence of synergistic peptide sequences on osteoblast adhesion, spreading, and function. Specifically, acrylated monomers were prepared that contained the peptide sequence, Arg-Gly Asp (RGD), as well as monomers with RGD plus its synergy site, Pro-His-Ser-Arg-Asn (PHSRN), linked via a polyglycine sequence to recapitulate the native spacing of fibronectin. The colocalized RGD-PHSRN sequence improved osteoblast adhesion, spreading, and focal contact formation when compared to RGD alone. In addition, proliferation, metabolic activity, and levels of alkaline phosphatase production, a common marker for osteoblast function, were statistically higher for the colocalized peptide sequences at 1 day, 1 week, and 2 weeks, when compared to control surfaces. Interestingly, increases were not observed in all areas of cell function, as extracellular matrix (ECM) production was the lowest on gels functionalized with the colocalized peptide sequence. This result was attributed to strong receptor-ligand interactions initiating signal transduction cascades that down-regulate ECM production. PMID- 15792549 TI - Stimulation of osteoblast responses to biomimetic nanocomposites of gelatin hydroxyapatite for tissue engineering scaffolds. AB - Collagen-derived gelatin/hydroxyapatite (HA) nanocomposites were biomimetically synthesized for hard tissue engineering scaffold. In vitro osteoblastic cellular responses to the nanocomposites were assessed in comparison with those conventionally mixed gelatin-HA composites. A three-dimensional culture method involving floating cells in a culture medium was introduced to assist in the initial attachment of the cells to the scaffolds, and the proliferation and differentiation behaviors of the cells were examined. The osteoblastic MG63 cells attached to the nanocomposites to a significantly higher degree and subsequently proliferated more. The alkaline phosphatase (ALP) activity and osteocalcin produced by the cells were significantly higher on the nanocomposite scaffolds than on the conventional composite scaffolds. These improved cellular responses on the nanocomposites are considered to result from the increased ionic release and serum protein adsorption on the nanocomposites, which was derived from the different structural and morphological characteristics, i.e., the nanocomposite scaffolds retained less-crystallized and smaller-sized apatite crystals and a more well-developed pore configuration than the conventional ones. Based on these findings, the biomimetically synthesized nanocomposite scaffolds are believed to be potentially useful in hard tissue regeneration and tissue engineering fields. PMID- 15792550 TI - Bone tissue engineering on amorphous carbonated apatite and crystalline octacalcium phosphate-coated titanium discs. AB - Poor fixation of bone replacement implants, e.g. the artificial hip, in implantation sites with inferior bone quality and quantity may be overcome by the use of implants coated with a cultured living bone equivalent. In this study, we tested, respectively, amorphous carbonated apatite (CA)- and crystalline octacalcium phosphate (OCP)-coated discs for their use in bone tissue engineering. Subcultured rat bone marrow cells were seeded on the substrates and after 7 days of culture, the implants were subcutaneously implanted in nude mice for 4 weeks. After 7 days of culture, the cells had formed a continuous multi layer that covered the entire surface of the substrates. The amount of cells was visually higher on the crystalline OCP-coated discs compared to the amorphous CA coated discs. Furthermore, the amorphous CA-coated discs exhibited a visually higher amount of mineralized extracellular matrix compared to the crystalline OCP coated discs. After 4 weeks of implantation, clear de novo bone formation was observed on all discs with cultured cells. The newly formed bone on the crystalline OCP-coated discs was more organized and revealed a significantly higher volume compared to the amorphous CA-coated discs. The percentage of bone contact with the discs was also significantly higher on the OCP-coated discs. Overall, the results suggest that a crystalline OCP coating is more suitable for bone tissue engineering than an amorphous CA coating. PMID- 15792551 TI - Thrombospondin 1 as possible key factor in the hemocompatibility of endocoronary prostheses. AB - Intracoronary stenting has markedly improved the patency of native coronary arteries after percutaneous transluminal coronary angioplasty (PTCA). Advances in stent technology and design, including drug releasing stents, have contributed to reduce the long-term restenosis rate. However, stenosis caused by neointimal hyperplasia, vascular remodeling and thrombosis is still a major problem after endocoronary stent procedures. This study focuses on differential gene expression of circulating peripheral blood cells after 90 min exposure to stents to search for initially activated cellular pathways, which may foster restenosis. Fresh human whole blood (1 IU heparin/ml), taken from non-medicated healthy volunteers, was incubated under flow conditions in an in vitro closed-loop stent-testing model (modified Chandler-Loop). Differential gene expression compared to resting conditions and to the experimental controls was investigated by a DNA-microarray technique encoding for over 17,000 genes simultaneously. As expected, a large variety of genes showed differential gene expression. Interestingly, Thrombospondin 1 (TSP-1), which plays a key role in initial immune defense, was found to be the most markedly up-regulated gene. We propose TSP-1 expression as an early indicator for the activation of immune responses following intracoronary stenting. After clarifying the participation of TSP-1 in vivo, future studies will therefore focus on TSP-1 as a potential prognostic factor, which may also help to develop and control new surface materials with an improved biocompatibility. PMID- 15792553 TI - Sustained release of human growth hormone from in situ forming hydrogels using self-assembly of fluoroalkyl-ended poly(ethylene glycol). AB - Poly(ethylene glycol)s modified with fluorocarbon end groups are capable of in situ transition from an injectable liquid to a viscoelastic hydrogel by hydrophobic interaction of the end groups; this class of materials is useful for a variety of biomedical applications, including sustained protein release. The hydrogel state can be transformed into an injectable state by the addition of a toxicologically acceptable organic solvent, such as N-methyl pyrrolidone; after injection, this solution quickly returns to a gel state by diffusion of the water miscible organic solvent into the surrounding environment. In vitro characterization of sustained release of human growth hormone (hGH) using this injectable depot shows that hGH remains stable inside the hydrogel formed, and demonstrates more than 2 weeks of prolonged release of hGH complexed with Zn(2+) ions without protein aggregation or initial burst. PMID- 15792552 TI - Effectiveness of hydroxyapatite-vancomycin bone cement in the treatment of Staphylococcus aureus induced chronic osteomyelitis. AB - In the field of local application of antimicrobials, a number of novel drugs and/or new drug delivery systems have been developed in recent years. The present study aimed to investigate hydroxyapatite cement (HAC) as a carrier for vancomycin in the treatment of chronic osteomyelitis due to Staphylococcus aureus strains with various mechanisms of resistance. The release of vancomycin from standard test cylinders was determined in vitro and the efficacy of the delivery system was measured in vivo using a rabbit model of chronic osteomyelitis. First, powdered HAC was mixed with vancomycin at 80, 160 and 240 mg/g. After hardening, formed cylinders were eluted in phosphate buffer and antibiotic release was measured by agar diffusion. High levels of release (1512+/-318 to 1937+/-336 microg/ml) were obtained for 12 to 20 days depending on the dosage of vancomycin. Additionally, bone infection was induced in the tibia of 30 New Zealand white rabbits by injecting either a methicillin-resistant S. aureus strain (MRSA) or a S. aureus strain with a small colony variant (SCV) phenotype. After 3 weeks (chronic infection), all animals were treated by debridement. Moreover, group 1 (challenged with SCVs) and group 2 (challenged with MRSA) were treated by filling the marrow with HAC alone, whereas in groups 3 (SCVs) and 4 (MRSA) the marrow was filled with HAC/vancomycin (160 mg/g). After 6 weeks all animals were sacrificed. At 3 weeks, pathogens were detected in 24 of 30 animals. All swabs of the control groups, positive for S. aureus on day 21, were also positive on day 42 and S. aureus strains recovered were shown to be clonal to the strains used for induction of osteomyelitis. By contrast, no growth was found in the treatment group following 7 days of incubation in BHI bouillon. HAC/vancomycin-treated animals showed no histological evidence of infection on day 42. In the other groups, different stages of chronic osteomyelitis were found histologically. No local or systemic side effects due to HAC or vancomycin were seen. HAC is an effective carrier material for antibiotic compounds even in refractory infections due to MRSA or S. aureus SCVs. PMID- 15792554 TI - [Nerve stimulation, nerve stimulator and the reasoned multiple nerve stimulation technique...]. PMID- 15792555 TI - [The neurostimulator for brachial plexus blockade by the axillary approach: a metaanalysis on its efficacy to increase the success rate]. AB - OBJECTIVE: To evaluate the effectiveness of the neurostimulator to improve the success rate of a brachial plexus blockade by the axillary approach. STUDY DESIGN: Metaanalysis. METHODS: Ten prospective randomized studies comparing a neurostimulating technique (NST) of brachial plexus blockade performed by the axillary approach to another technique were kept for analysis. A successful block was defined as blockade of the four following nerves: radial, median, ulnar and musculocutaneous. Data were entered on an intention to treat basis and were analyzed with a random model. A p<0.05 was considered significant. RESULTS: Data were heterogenous (p < 0.001) and the Cochran-Mantel-Haenzel test showed that the neurostimulator does not improve the success rate of brachial plexus blocks performed by the axillary approach when all available studies are considered as a whole (p = 1). For the subgroup of studies where four nerves were stimulated however, the NST decreases the incidence of failed block by 25% (95% CI = 7-42%) (p < 0.0001). There were 12 patients with systemic signs of local anesthetics toxicity when a NST was used compared to 28 when it was not (p = 0.04), relative risk of 0.7 (95% CI= 0.6-0.8). CONCLUSION: This study shows that the neurostimulator improves the success rate of brachial plexus blocks performed by the axillary approach only when three nerves or more are stimulated and its use decreases the incidence of systemic local anaesthetics toxicity. PMID- 15792556 TI - [Regionalization of obstetric services and anaesthetic workload in a tertiary level perinatal unit]. AB - OBJECTIVE: Evaluation of anaesthetic workload associated with care of high-risk pregnant women (i.e. patients transferred to a referral perinatal center). STUDY DESIGN: A case-control prospective study. METHODS: During a three-month period, 63 women with in-utero transfer and their control counterparts (63 normal pregnancies) were prospectively followed until discharge. RESULTS: At inclusion, high-risk patients (n = 63) were more frequently scored ASA 2 or more (21 vs 0%, p < 0.05). ASA score had increased at delivery in 8% of high-risk women and in none of controls. ICU admission (4 vs none), Caesarean delivery (57 vs 11%), preanaesthetic visit at night (41 vs 0%) and general anaesthesia (27 % versus none) were all more frequent in high-risk parturients (p < 0.05). CONCLUSION: Comparison of the workload associated with high-risk patients and normal pregnant women confirms the need for increased anaesthetic staffing in referral perinatal centers and provides a basis for better understanding the distribution of anaesthetic requirements in each perinatal network. PMID- 15792557 TI - [Unilateral spinal anaesthesia in elderly patient for hip trauma: a pilot study]. AB - INTRODUCTION: Fractured neck of femur is encountered more frequently as life expectancy increases. All anaesthetic techniques aim to avoid deleterious fall in arterial blood pressure. The haemodynamic effects of hypobaric unilateral spinal anaesthesia have been rarely assessed in patients over 80 year's old. This study aims to do that in a prospective manner. PATIENTS AND METHODS: Twenty-five patients were enrolled. Following a preload with HES 240/0.576 % (Hesteril) (5 ml/kg) and the administration of an iliofascial block, patients were placed in the lateral decubitus position, operative side uppermost. 3.5 ml of 0.12% hypobaric bupivacaine was administered intrathecally at a rate of 0.25 ml/second. Patients were kept in position for 15 minutes. Systolic, mean and diastolic arterial pressures, heart rate, SaO2 % and ephedrine consumption were recorded at five minutes intervals as was the rate of onset, height and duration of sensory and motor block and extent of bilateralization. Patient and surgeon satisfaction scores were also recorded. RESULTS: No significant changes in systolic, mean and diastolic pressures, or SaO2 % occurred. Median onset times of sensory and motor block were 8+/-5 and 16+/-7 minutes on the operative side and 30+/-15 and 36+/-15 minutes on the contralateral side in those with bilateralization, respectively. The maximum height of sensory block was T6 for sept patients, T8 for huit patients and T8-T10 for the remainder. Mean duration of sensory and motor block was 134+/-26 and 119+/-24 minutes on the operative side and 100+/-26 and 98+/-25 minutes on the contralateral side, respectively. In 12 patients (48%) bilateralization of their block occurred. Patients and surgeons rated the technique highly. CONCLUSION: Hypobaric unilateral spinal anaesthesia is a simple technique, produces satisfactory operative conditions and induces very little haemodynamic change in the elderly population. PMID- 15792558 TI - [Differential modulation of TLR2 and TLR4-induced TNF production by murin haemorrhagic shock]. AB - OBJECTIVE: To investigate the influence of haemorrhagic shock in mice on ex vivo TNF production by whole blood cells (WBC) stimulated through Toll-like receptors (TLR) 4 and 2. STUDY DESIGN AND ANIMALS: Experimental study using BALB/c male mice. METHODS: Haemorrhage (0,026+/-0,003 ml/g) by transparietal cardiac puncture under general anaesthesia. Measurement of left intraventricular pressure through a direct subcostal cardiac puncture. Possible restitution of shed blood volume (SBV) in retroorbital venous plexus, 60 minutes following haemorrhage. Lethal exsanguination 120 minutes following general anaesthesia (Control group), cardiac puncture (Sham group), blood sample (Haemorrhage group), or 60 minutes following SBV retransfusion (SBV group). Cultures (24 hours) of whole blood from the exsanguination, alone or with Escherichia coli endotoxin (LPS, TLR 4) or with heat-killed Staphylococcus aureus Cowan (SAC, TLR 2). Assessment of TNF levels in the cultures supernatant (Elisa). RESULTS: Hemorrhage (approximately 30% of calculated blood volume) resulted in arterial hypotension (-50%) which was reversed by SBV retransfusion. TNF production by LPS-stimulated WBC was reduced by haemorrhage (approximately -50%) with or without SBV retransfusion. TNF production by SAC-stimulated WBC remained unchanged. CONCLUSION: The reduction of proinflammatory cytokines production by WBC stimulated with pathogen-associated molecular patterns is not a generalized phenomenon following murin haemorrhagic shock. It depends on the used stimulus and studied signalling pathways. PMID- 15792559 TI - [Interest of mannequin based simulator to evaluate anaesthesia residents]. AB - OBJECTIVE: The aim of this study was to test simulator validity to evaluate the ability of anaesthesia residents to solve two simulated scenarios. STUDY DESIGN: Monocentre, prospective, randomized study. POPULATION: Anaesthesia residents. METHODS: All anaesthesia residents were invited to participate into the study but were free to decline to take part. The authors developed grading forms to evaluate preoperative preparation of anaesthesia room and two simulated scenarios which had been previously validated. All residents were evaluated on the preoperative preparation of anaesthesia room. A randomization was performed to select half of the residents to be tested on one of the simulated scenario. Two experienced anaesthesiologists scored the residents' performance. At the end of the simulated session, residents rated the realism of the scenarios. RESULTS: Among 72 training residents in our institution, 48 participated with 24 beginning and 24 advanced residents. Median scores were similar between beginning (first and second year) and advanced residenced (third and fourth year) for the preoperative preparation of anaesthesia room (17 vs 17 for a maximal score of 25) while scores tended to be higher in advanced residents for simulated scenarios (scenario 1 [34 vs 19 for a maximal score of 55; p = 0.0009], scenario 2 [17 vs 13 for a maximal score of 45; p = 0.58]). However, numerous management errors were observed and some of them did not improve with training. Anaesthesia residents rated the simulator scenarios as realistic. CONCLUSION: This study suggests that mannequin-based simulator appears as a reliable and valid tool to test the performance of anaesthesia residents during critical situations. PMID- 15792561 TI - [Extensive mesenteric venous thrombosis treatment by regional thrombolysis]. AB - Two cases of mesenteric venous thrombosis with portal extension are reported. The first patient was treated right away by local intra-arterial thrombolysis, the second one benefited from local venous thrombolysis immediately after intestinal resection. No significant complication was observed. PMID- 15792560 TI - [Important hypercalcaemia due to hyperparathyroidism induced by lithium]. AB - Lithium treatment, which is still extensively used in bipolar affective disorders, may give rise to hypercalcaemia induced by hyperparathyroidism. We present a patient of 50-year-old treated with lithium for 19 years for bipolar illness and who developed an important hypercalcaemia. After symptomatic treatment of the hypercalcaemia and extrarenal dialysis the clinical evolution was favorable but measurements of serum calcium and parathormon showed that he had developed hyperparathyroidism. Neck exploration was performed and parathyroid adenomas, which had been detected by scintigraphy was removed. The lithium treatment expose to many side effects. Among other biologically and clinically important effects of lithium the possible induction of hyperparathyroidism was first suggested in 1973. Since, 1973, since about forty case reports have been described. Few cross-sectional studies show a relationship of lithium to hyperparathyroidism. Unusual metabolic features are associated with hyperparathyroidism and long-term lithium treatment: low urinary calcium excretion, normal urinary cyclic AMP excretion. The mechanism probably results from lithium linking with the calcium receptor on the parathyroid and then stimulating PTH secretion. The cessation of lithium therapy does not lead to normocalocaemia and a parathyroidectomy is usually indicated. PMID- 15792562 TI - [Epidemiology of catheter-related infections in intensive care unit]. AB - Catheter-related infections remain an important cause of nosocomial infection in the ICU. They include colonization of the device, exit-site infection and catheter-related bloodstream infection with or without bacteraemia. Data from clinical studies and surveillance networks should be compared cautiously due to important methodological differences and wide variations of device-utilization ratio between units or countries. In France, two regional networks (C-CLIN Paris Nord and C-CLIN Sud-Est) produced comparable and reproducible results. Colonization represents five-six cases per 1000 catheter-days and bacteraemia represents one case per 1000 catheter-days. Incidence rates from North American studies are usually four to five times higher. Numerous risk factors have been identified. Some of them could be used to stratify patients according to risk of catheter-related infection and to allow more valid comparison between ICU's performances. Participation of French ICUs to the recent national surveillance networks (REA RAISIN and REACAT RAISIN) should be encouraged. PMID- 15792563 TI - [Catheter-related infections: microbiology]. AB - Coagulase negative staphylococci, Staphylococcus aureus and Pseudomonas sp. are the most frequent microorganisms responsible for catheter-related infections. A relative frequency of microorganisms varies according to the countries, microenvironment and outbreaks of multiresistant bacterias. Infections due to fungi, S. aureus and Pseudomonas sp. are associated with the more severe complications. Recent data suggest that chlorhexidine, either used for cutaneous antisepsis or for catheter impregnation decreases infections due to gram positive cocci. Ecological data should be taken into account when deciding a probabilistic treatment in case of suspicion of catheter-related infection. PMID- 15792564 TI - [On what grounds assessing the diagnosis of catheter-related infection?]. AB - Clinical assessment of catheter-related infection is difficult. Microbiological techniques have not always been clearly evaluated. Indirect microbiological techniques, leaving the catheter on site are reserved to well tolerated infections. In case of catheter withdrawal, quantitative culture of the endovascular tip of the catheter should be preferred. PMID- 15792565 TI - [Catheter-related infections: how to reduce the risk?]. AB - Subcutaneous tunnelling of short-term catheters and the choice of site of catheter insertion are the two factors which influence the risk of catheter related infection. Catheters inserted into a femoral vein have been associated with a higher risk of infection than those inserted into a subclavian vein. However, a meta-analysis comparing the internal jugular and subclavian sites did not report any difference. Regarding tunnelling, two randomized studies showed a benefit in terms of infection rates for the jugular and femoral sites when the blood sampling was not performed via the catheters. For the subclavian site tunnelling does not seem to bring any advantage compared to conventional insertion. PMID- 15792566 TI - [Clinical use of antiseptic and antibiotic impregnated catheters]. AB - In order to decrease the risks of colonisation and infection of central venous catheters, their impregnation has been suggested using anti-infectious agents (antiseptic, antibiotic, silver). Many methodological flaws and contradictory results have been reported in the published trials. The use of impregnated catheter should only be discussed when the issues of anatomic site and type of insertion of the catheter have been adequately addressed. PMID- 15792567 TI - [Should we use catheter replacement with the aid of wire introducer?]. AB - Two types of catheter replacement with the help of wire introducer are reported: systematic scheduled replacement and replacement in case of suspicion of catheter related infection. Guidelines do not recommend systematic scheduled replacement of the catheters. In case of suspicion of catheter infection, French consensus guidelines allow the use of wire introducer in the absence of local risk and of signs of severity. The American guidelines do not recommend the catheter change over guidewire in this setting. PMID- 15792568 TI - [Is there any microbiologically-based specificity of the management of catheter related infections?]. AB - After microbiological identification, several points are to be taken into account: microbiological results, clinical signs of severity, catheter withdrawal or not. French and American recommendations are very different for coagulase negative staphylococci. Catheter withdrawal and antibiotic treatment are recommended by all the authors for coagulase positive staphylococci infections. There is no a clear guideline or studies on Enterobacteriaceae or Pseudomonas infections. Catheter removal and antifungal treatment are recommended for Candida infection. PMID- 15792569 TI - [Management of catheter-related infections: factors of decision in treatment]. AB - Whenever a catheter related infection is suspected, two main issues must be addressed: should the catheter be removed and should an empiric antibiotic treatment be prescribed? These questions are still under debate. Some differences are found between French and American guidelines. Main factors leading to decision about these questions are: severity of the sepsis, microorganisms involved and kind of patients we are dealing with. PMID- 15792570 TI - [Lingual haematoma after thrombolysis for acute myocardial infarction]. PMID- 15792571 TI - [Tear of a disposable laryngeal mask airway]. PMID- 15792572 TI - [Neuraxial anaesthesia and hereditary motor and sensory neuropathies]. PMID- 15792573 TI - [Paracervical block for hysteroscopy in patient with spinal muscular atrophy]. PMID- 15792574 TI - [Paracervical block]. PMID- 15792575 TI - [Updating of the 12th consensus conference of the Societe de Reanimation de langue francaise (SRLF): catheter related infections in the intensive care unit]. AB - Concerning catheter-related infections, many studies have been conducted until the last consensus conference of the Societede reanimation de langue francaise (SRLF) in 1994. This text is the synthesis of the considerable amount of work performed by the experts of the society to review recent studies. The experts' texts as well as extensive bibliography are available at http://www.srlf.org. PMID- 15792576 TI - [Report on the 18th meeting of the club d'anesthesie-reanimation en ORL (Carorl), Poitiers, November 26, 2004]. PMID- 15792577 TI - Effect of prefermentation on denitrifying phosphorus removal in slaughterhouse wastewater. AB - An anaerobic-anoxic sequencing batch reactor (A2 SBR) coupled with a fixed-bed nitrification reactor for simultaneous carbon, nitrogen and phosphorus removal was evaluated using slaughterhouse wastewater. Whereas the treatment could not be successfully carried out on the raw wastewater, the process showed very good nutrient removal performances after prefermentation. The removals of COD, N-NH4 and P-PO4 achieved were 99%, 85% and 99%, respectively. The increase in volatile fatty acid (VFA) and phosphate concentrations in the effluent after prefermentation may explain the high levels of biological carbon, nitrogen and phosphorus removal observed. A simple prefermentation is, therefore, necessary but sufficient to ensure good performances of the denitrifying enhanced biological phosphorus removal (EBPR) process. PMID- 15792578 TI - Improving seedling vigour of indigenous medicinal plants with smoke. AB - The application of smoke and aqueous smoke solutions stimulates seed germination in a number of plant species. This study highlights the effects of aerosol smoke and smoke solutions on the germination and seedling vigour of three South African indigenous medicinal plants Albuca pachychlamys, Merwilla natalensis and Tulbaghia violacea. The vigour index of one-week-old seedlings of all three species examined was increased with the application of dry smoke and smoke extract dilutions, as compared to control treatments. Seedlings of A. pachychlamys germinated with smoke solutions showed a significant (p0.05) gain in bulb and leaf mass (27.9 and 197.6 mg respectively) compared to untreated seedlings (9.9 and 124.7 mg respectively) when grown in vitro for 75 days. The leaf mass of smoke solution-treated seedlings of T. violacea was significantly (p0.05) higher (120.4 mg) than that of untreated seedlings (47.6 mg). Subsequently, the height of seedlings in both species was also significantly (p0.05) greater. Seedlings germinated in water and then transferred to smoke solutions (1:2000) showed enhancement of some of the growth parameters studied. Albuca pachychlamys and T. violacea seeds exposed to aerosol smoke exhibited higher seedling survival percentages than from non-smoked seeds, while no significant effect was observed for M. natalensis seedlings. This investigation shows that the application of smoke technology can be adopted to produce high vigour seedlings. PMID- 15792579 TI - Biostabilization-biodrying of municipal solid waste by inverting air-flow. AB - The process of biodrying could be a good solution for municipal solid waste management, allowing the production of fuel with an interesting energy content. Previous work (Adani, F., Baido, D., Calcaterra, E., Genevini, P.L., 2002. The influence of biomass temperature on biostabilization-biodrying of municipal solid waste. Bioresource Technology 83 (3), 173-179) has indicated that appropriate management of the processing parameters (air-flow rate and biomass temperatures) could achieve biomass drying in very short times (8-9 days). However, the data of that work also evidenced that if the conditions do not consider pile turning, and the air-flow is always from one direction, temperature gradients arise within the biomass, resulting in a lack of homogeneity in the moisture and energy content of the final product. Therefore, a new laboratory study was conducted on municipal solid waste biodrying-biostabilization in an effort to obtain homogeneous final products. Our proposal to solve this lack of homogeneity is to periodically invert the air-flow direction. Thus, in line with a previous study, two trials, A and B, were carried out, dividing the biomass into three layers to study temperature and moisture gradients throughout the process, and a third trial (C) simulating air-flow inversion at regular intervals was introduced. The results suggest that the daily inversion of air-flow eliminates marked temperature differences and leads to a homogeneous final product. PMID- 15792580 TI - Management of Meloidogyne incognita on tomato by root-dip treatment in culture filtrate of the blue-green alga, Microcoleus vaginatus. AB - The nematicidal potential of culture filtrates of the blue-green alga, Microcoleus vaginatus (Cyanobacterium) was tested against Meloidogyne incognita on tomato in pots under greenhouse conditions. Prior to the transplantation of tomato seedling, roots were dipped in different concentrations (0.2%, 0.5%, 1%, 2%, 10%, 50% and 100%) of culture filtrate of M. vaginatus for 30 min. Root-dip treatment reduced the root galling and final population of M. incognita and increased vegetative growth of plants and root-mass production compared with the control. The beneficial effect of root-dip treatment increased with the increase in the concentration of culture filtrate. Root galling and final nematode populations were reduced by 65.9% and 97.5%, respectively when treated at the highest concentration. PMID- 15792581 TI - Physicochemical characterisation of residual hemicelluloses isolated with cyanamide-activated hydrogen peroxide from organosolv pre-treated wheat straw. AB - Seven residual hemicellulosic preparations (19.6-45.0% of the original hemicelluloses) were extracted from wheat straw pre-treated with various organic solvents using 1.8% H2O2-0.18% cyanamide at 50 degrees C and pH 10.0 for 4 h. Their chemical compositions and physicochemical properties were determined using GC, HPLC, GPC, FT-IR and 13NMR spectroscopy. The results indicated that all the residual hemicellulosic preparations were heteropolysaccharides containing xylose, glucose, arabinose, galactose, mannose, rhamnose and 4-O-methyl-alpha-D glucopyranosyluronic acid. The predominant monosaccharide was xylose, ranging between 67.7% and 81.9% of the total neutral sugars, composed mainly of L-arabino (4-O-methyl-D-glucurono)-D-xylan. The content of contaminant lignin in the isolated residual hemicelluloses was 2.89-5.31%. The Mw values of the two residual hemicellulosic preparations H6 and H7 (42,710 and 44,080 g mol-1, respectively) obtained from the aqueous-alcohol pre-treated straw were much higher than those of H1-H5 (12,980-15,950 g mol-1) extracted from the organic acid pre-treated straw. PMID- 15792583 TI - Degradation of lignin in pulp mill wastewaters by white-rot fungi on biofilm. AB - An investigation was conducted to explore the lignin-degrading capacity of attached-growth white-rot fungi. Five white-rot fungi, Phanerochaete chrysosporium, Pleurotus ostreatus, Lentinus edodes, Trametes versicolor and S22, grown on a porous plastic media, were individually used to treat black liquor from a pulp and paper mill. Over 71% of lignin and 48% of chemical oxygen demand (COD) were removed from the wastewater. Several factors, including pH, concentrations of carbon, nitrogen and trace elements in wastewater, all had significant effects on the degradation of lignin and the removal of COD. Three white-rot fungi, P. chrysosporium, P. ostreatus and S22, showed high capacity for lignin degradation at pH 9.0-11.0. The addition of 1 g l-1 glucose and 0.2 g l-1 ammonium tartrate was beneficial for the degradation of lignin by the white-rot fungi studied. PMID- 15792582 TI - A study of boron adsorption onto activated sludge. AB - Boron adsorption onto activated sludge was investigated using bench-scale reactors under simulated wastewater treatment conditions. Two experiments, continuous flow and batch, were performed. Boron concentrations were determined by means of inductively coupled plasma mass spectrometry. The results of the continuous-flow experiment indicated that a small amount of boron accumulated on the activated sludge and its concentration in the sludge depended on the nature of the biota in the sludge. Freundlich and Langmuir isotherm plots generated using the data from the batch experiment indicated that boron was adsorbed onto rather than absorbed into the sludge. The Freundlich constants, k and 1/n, were determined to be 26 mg/kg and 0.87. These values indicate that activated sludge has a limited capacity for boron adsorption and thus utilization of the excess sludge for farmland may not be toxic to plant at least boron concern. PMID- 15792584 TI - Activated carbon from char obtained from vacuum pyrolysis of teak sawdust: pore structure development and characterization. AB - The preparation of activated carbon from vacuum pyrolysis char of teak sawdust was studied and the results are presented in this paper. The effects of process variables such as temperature and activation time on the pore structure of activated carbons were studied. The activated carbon prepared from char obtained by vacuum pyrolysis has higher surface area and pore volume than that from atmospheric pyrolysis char. The BET surface area and pore volume of activated carbon prepared from vacuum pyrolysis char were 1150 m2/g and 0.43 cm3/g, respectively. PMID- 15792585 TI - Characterization of a cellulase-free, neutral xylanase from Thermomyces lanuginosus CBS 288.54 and its biobleaching effect on wheat straw pulp. AB - A xylanase purified from the thermophilic fungus Thermomyces lanuginosus CBS 288.54 was characterized and its potential application in wheat straw pulp biobleaching was evaluated. Xylanase was purified 33.6-fold to homogeneity with a recovery yield of 21.5%. It appeared as a single protein band on SDS-PAGE gel with a molecular mass of approx. 26.2 kDa. The purified xylanase had a neutral optimum pH ranging from pH 7.0 to pH 7.5, and it was also stable over pH 6.5 10.0. The optimal temperature of the xylanase was 70-75 degrees C and it was stable up to 65 degrees C. The purified xylanase was found to be not glycosylated. The xylanase was highly specific towards xylan, but did not exhibit other enzyme activity. Apparent Km values of the xylanase for birchwood, beechwood, soluble oat-spelt and insoluble oat-spelt xylans were 4.0, 4.7, 2.0 and 23.4 mg ml-1, respectively. The potential application of the xylanase was further evaluated in biobleaching of wheat straw pulp. The brightness of bleached pulps from the xylanase pretreated wheat straw pulp was 1.8-7.79% ISO higher than that of the control, and showed slightly lower tensile index and breaking length than the control. Although chlorine consumption was reduced by 28.3% during bleaching, the xylanase pretreated pulp (15 U g-1 pulp) still maintained its brightness at the control level. Besides, pretreatment of pulp with the xylanase was also effective at an alkaline pH as high as pH 10.0. PMID- 15792586 TI - Optimization of medium composition for alkali-stable xylanase production by Aspergillus fischeri Fxn 1 in solid-state fermentation using central composite rotary design. AB - Response surface methodology and central composite rotary design (CCRD) was employed to optimize a fermentation medium for the production of alkali-stable cellulase-free xylanase by Aspergillus fischeri in solid-state fermentation at pH 9.0 with wheat bran as substrate. The four variables involved in this study were sodium nitrite, potassium dihydrogen phosphate, magnesium sulphate and yeast extract. The statistical analysis of the results showed that, in the range studied, only sodium nitrite had a significant effect on xylanase production. The optimized medium containing (in g/l) NaNO(2)-7.0, K2HPO(4)-1.0, MgSO(4)-0.5 and yeast extract-5.0 resulted in 1.9-fold increased level of alkali-stable xylanase (1024 U/g wheat bran) production compared to initial level (540 U/g) after 72 h of fermentation, whereas its value predicted by the quadratic model was 931 U/g. The level of protease activity was considerably decreased in optimized medium, thus helping to preserve the xylanase activity and demonstrating another advantage of applying statistical experimental design. PMID- 15792587 TI - Concurrent sorption of Zn(II), Cu(II) and Co(II) by Oscillatoria angustissima as a function of pH in binary and ternary metal solutions. AB - This paper reports biosorption of Zn(II), Cu(II) and Co(II) onto O. angustissima biomass from single, binary and ternary metal solutions, as a function of pH and metal concentrations via Central Composite Design generated by statistical software package Design Expert 6.0. The experimental design revealed that metal interactions could be best studied at lower pH range i.e. 4.0-5.0, which facilitates adequate availability of all the metal ions. The sorption capacities for single metal decreased in the order Zn(II)>Co(II)>Cu(II). In absence of any interfering metals, at pH 4.0 and an initial metal concentration of 0.5 mM in the solution, the adsorption capacities were 0.33 mmol/g Zn(II), 0.26 mmol/g Co(II) and 0.12 mmol/g Cu(II). In a binary system, copper inhibited both Zn(II) and Co(II) sorption but the extent of inhibition of former was greater than the latter; sorption values being 0.14 mmol/g Zn(II) and 0.27 mmol/g Co(II) at initial Zn(II) and Co(II) concentration of 1.5 mM each, pH 4.0 and 1mM Cu(II) as the interfering metal. Zn(II) and Co(II) were equally antagonistic to each others sorption; Zn(II) and Co(II) sorption being 0.23 and 0.24 mmol/g, respectively, at initial metal concentration of 1.5 mM each, pH 4.0 and 1mM interfering metal concentration. In contrast, Cu(II) sorption remained almost unaffected at lower concentrations of the competing metals. Thus, in binary system inhibition dominance observed was Cu(II)>Zn(II), Cu(II)>Co(II) and Zn(II) approximately Co(II), due to this the biosorbent exhibited net preference/affinity for Cu(II) sorption over Zn(II) or Co(II). Hence, the affinity series showed a trend of Cu(II)>Co(II)>Zn(II). In a ternary system, increasing Co(II) concentration exhibited protection against the inhibitory effect of Cu(II) on Zn(II) sorption. On the other hand, the inhibitory effect of Zn(II) and Cu(II) on Co(II) sorption was additive. The model equation for metal interactions was found to be valid within the design space. PMID- 15792588 TI - Growth and productivity assessment of Casuarina glauca Sieb. ex. Spreng on sodic soil sites. AB - Casuarina glauca is an introduced tree species to India. It is one the few species that can grow successfully in salt effected sites. C. glauca was investigated for biomass production at highly sodic soil sites (pH 8.6-10.5); it was found to be promising in terms of growth and productivity. Average height of plants in an 8-yr-old trial of Casuarina glauca was 1033.3+/-270 cm, however diameter at breast height (dbh) remained at 8.59+/-2.0 cm with a basal area of 6.68+/-1.6 cm2 per plant. High plant survival and establishment (75%) indicate its potential for afforestation of degraded soil sites. Linear regression equations (Y=a+bx) were developed to predict biomass of standing stocks defining relationships between growth parameters (x) as independent variable and productivity (Y) as dependent variable. Both uni-factor equations based on one independent variable (height or diameter alone), and multifactors involving both height and diameter together (d2h) were derived to predict biomass of different plant components. Coefficient of correlation and regression coefficients were found to be highly significant (p<0.001) in all the equations irrespective of independent variable such as plant height, diameter or both (d2h). There were marginal differences in r2 value (0.78-0.8) among equations derived by using d2 or d2h as independent variables. Height alone had relatively poor functional correlation with yield (r2=0.45). Accordingly, uni-factor linear equations with diameter (d2) were used for computing stand biomass with reasonably good accuracy. At the age of 8 years, stand productivity was 68.2 t ha-1 (oven dry biomass) out of which relatively a very high proportion (80.3%) of biomass was allocated to stem wood (54.8 t ha-1). Both branch wood (8.4 t ha-1) and leaves (5 t ha-1) contributed marginally. Casuarina glauca can be recommended as a promising species for biomass production on sodic soil sites as is evident from its performance. PMID- 15792589 TI - Comparison of different microbial biomass and activity measurement methods in metal-contaminated soils. AB - The aims of this study were: (1) to compare different microbial methods of detecting the effects of heavy metals on the functioning of the soil ecosystem; and (2) to evaluate the effect of incubation on microbial biomass and microbial activity in soils that were not pre-incubated after sampling in order to determine their suitability for measuring the effects of heavy metals on the soil microbial ecosystem. The microbial biomass methods (included: biomass C, N and ninhydrin-N by fumigation-extraction (FE); substrate-induced respiration (SIR); soil ATP content and microbial activity as evolved CO2-C and arginine ammonification. All were tested in soils from the Woburn Market Garden Experiment. Due to past sludge application the soils contained, Zn, Cu or Ni at around current European Union upper limits and Cd at up to three times the limit. The amount of microbial biomass in metal-contaminated soils was about half of that found in soils from the experiment that received uncontaminated organic manure or inorganic fertilizer. The amount of biomass measured by FE and soil ATP content in incubated soils showed little change over 20 days incubation. However, SIR measurements were statistically affected over the first few days of incubation. The rates of arginine ammonification were higher in this order: farmyard manure (FYM)>inorganic fertilizer>sewage-sludge throughout the incubation. However, the evolved CO2-C rates were not significantly different among the treatments. Discriminant analysis confirmed smaller amounts of biomass in the metal-contaminated soils than in the other treatments. Linked properties, such as relationships between biomass and soil organic matter, or biomass specific respiration rates, may provide "internal control" which may help overcome problems of establishing suitable control, or comparative measurements, when moving from experimental to natural environments. PMID- 15792590 TI - Effect of antibiotics on growth and laccase production from Cyathus bulleri and Pycnoporus cinnabarinus. AB - The effect of nine different antibiotics (chloramphenicol, ampicillin trihydrate, kanamycin A monosulfate, neomycin sulfate, erythromycin, thiostrepton, tetracycline, apramycin sulfate and streptomycin sulfate) on growth and laccase production from Cyathus bulleri and Pycnoporus cinnabarinus has been investigated. All the antibiotics tested at a concentration of 200 mg/l affected the fungal growth, release of protein and laccase production to different extent. Inhibition in fungal growth was found to be positively correlated with increase in laccase production. Interestingly, apramycin sulfate inhibited biomass production (14.9-26.2%), nevertheless, it stimulated maximum laccase production (18.2 U/ml) in both the fungi. Increasing concentrations of apramycin sulfate enhanced laccase production from P. cinnabarinus but not from C. bulleri. PMID- 15792591 TI - Soybean milk residue ensiled with peanut hulls: fermentation acids, cell wall composition, and silage utilization by mixed ruminal microorganisms. AB - Preservation of soybean milk residue (SMR) by ensiling with peanut hulls (PEH) and subsequent utilization of silage by mixed ruminal microorganisms were investigated. Treatments were combinations of SMR with PEH at the following ratios: 100:0, 78:22, 71:29, and 60:40 (fresh weight basis). After eight weeks of ensiling, silage lactic acid, crude protein, ether extract, and non-fiber carbohydrates were highest when SMR was ensiled alone and reduced as amounts of SMR decreased. Similar trends were observed for silage in vitro dry matter digestibility, and gas and volatile fatty acid production by ruminal microorganisms. Conversely, silage pH, dry matter, neutral detergent fiber, acid detergent fiber, cellulose, and lignin increased accordingly. The ensiling treatment appeared to alter silage cell wall composition. In particular, silage treated with PEH at the low level (78:22) resulted in reduced fiber contents and lignification. The silage (SMR:PEH=78:22) had enhanced efficiency of both silage fermentation and in vitro ruminal fermentation pattern. PMID- 15792592 TI - The chemical composition of the uropygial gland secretion of rock dove Columba livia. AB - The uropygial gland is a holocrine secretory gland of birds. The lipid and the waxy sebum that the gland secretes is coated on the beak and transferred to the plumage in preening. The composition of the gland secretions from birds of different species has been determined, but little is known about the lipids of the secretion of the gland of the rock dove Columba livia. The amount of secretion, the total lipid content and the fatty acids composition of the secretion of C. livia captured in the nonbreeding season was reported. The mean amount of the secretion within the gland was 30 mg; the mean lipid content of the secretion was 0.385 mg/mg of secretion, which was equivalent to approximately 38% of the secretion. The weight of the secretion relative to gland weight was 32%. If we assume that the amount of the gland secretion constitutes a valid parameter to determine the degree of the gland development, our results indicate that the physiological role of the gland does not depend upon gland mass (GW); the rock dove in particular has a small gland, but its secretion represented 32% of the gland's mass. The composition of the lipids extracted from the gland secretion consisted of C14 to C20 fatty acids, most of them were unsaturated. The secretion of the gland contained approximately 59% of unsaturated fatty acids with a prevalence of oleic acid (37%) and a low content of linoleic (6%) and arachidonic acids (7%). The saturated long chain fatty acids were mainly 14:0, 16:0 and 18:0 in a percentage of approximately 34%. No sexual differences were found in any of the evaluated parameters. PMID- 15792593 TI - Erythrocyte senescence and haematological changes induced by starvation in the neotropical fish traira, Hoplias malabaricus (Characiformes, Erythrinidae). AB - Adult specimens of traira (Hoplias malabaricus Bloch) were subjected to long-term starvation (30 to 240 days) and re-fed for 30 days after 90 and 240 days of food deprivation. Counting of immature erythrocytes in peripheral blood showed that erythropoiesis decreased significantly during the first 30 days of food deprivation. The results suggest that a process of senescence takes place in the pre-existent red blood cells and that the cells are not replaced during starvation. After 240 days of starvation, H. malabaricus had a significantly reduced number of red blood cells, causing changes in hematocrit and blood indices (mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration). Furthermore, during this period, the fish presented leukopenia (lymphocytopenia) and thrombocytopenia. After re-feeding, the number of leukocytes and thrombocytes recovered, but the red blood cell number remained reduced and there was a significant increase in abnormal red cell nuclei. PMID- 15792594 TI - Cardiovascular changes and catecholamine release following anaesthesia in Chinook salmon (Oncorhynchus tshawytscha) and snapper (Pagrus auratus). AB - We investigated recovery from anaesthesia in Chinook salmon (Oncorhynchus tshawytscha) with and without surgery. Fish either underwent light sedation on exposure to 60 ppm AQUI-S or surgical depth anaesthesia with 120 ppm AQUI-S. Surgical depth anaesthesia experiments were replicated using New Zealand snapper (Pagrus auratus). During light sedation, there was no evidence of catecholamine release in salmon despite changes in heart rate and blood pressure. Following surgical anaesthesia both salmon and snapper released high concentrations of catecholamines into the circulation. Plasma half-life of adrenaline in salmon was 9.3+/-0.7 min (n = 7) and in snapper was 4.4+/-3.3 min (n = 7). There was no further release of catecholamines, despite attempts by both species to escape their enclosures. Though clearance of the catecholamines was rapid, the cardiovascular effects of anaesthesia were prolonged. Dorsal aortic blood pressure (P(DA)) and heart rate (HR) were high following anaesthesia, falling by 60 min in the 60 ppm exposed salmon but remaining high in the 120 ppm group. Following anaesthesia ventral aorta blood pressure (P(VA)) in snapper was positively correlated with HR, as was P(DA) and haematocrit in salmon. Recovery of cardiovascular control processes is prolonged in recovery from anaesthesia if the fish become hypoxic. PMID- 15792595 TI - Energy metabolism and nutrient oxidation in young pigs and rats during feeding, starvation and re-feeding. AB - The investigation included individual measurements of energy metabolism and oxidation of nutrients in 12 castrated male pigs (Sus scrofa) (20-40 kg) and 12 male rats (Rattus norvegicus) (65-105 g). Measurements were carried out in 5-6 days balance periods with ad libitum feeding, followed by 3-4 days of starvation and 4 days of re-feeding. O2 consumption and CO2 production were measured by open air-circuit respiration units. In the feeding period, protein retention in relation to metabolic live mass (kg(0.75)) was identical for pigs and rats, while there was a tendency of a higher fat retention in pigs than in rats. A substantial part of digested carbohydrate was not oxidized, but transferred to fat metabolism without significant differences (P > 0.05) between pigs and rats (18% vs. 22%). During starvation, nitrogen excretion in urine decreased to 226 mg/kg(0.75) in pigs and to 429 mg/kg(0.75) in rats, indicating a lower rate of body protein degradation in pigs. Heat production was reduced to 592 and 338 kJ/kg(0.75), while the contribution of heat from oxidation of protein (OXP), carbohydrate (OXCHO) and fat (OXF) showed the same pattern for pigs and rats during all periods. Heat production during feeding and re-feeding was covered by OXP+OXCHO with no OXF and reversibly after 2 days of starvation by OXP+OXF with no OXCHO. The rat may be a suitable model for pigs regarding general patterns of quantitative nutrient partition, but any direct application of results measured with rats to pigs shall be taken cautiously, keeping in mind that modern pigs have been selected for a high growth rate and protein deposition which has not been the case for the laboratory rat. PMID- 15792596 TI - Reduction of the Pa(CO2) set point during hyperthermic exercise in the sheep. AB - In animals that rely on the respiratory system for both gas exchange and heat loss, exercise can generate conflict between chemoregulation and thermoregulation. We hypothesized that in panting animals, hypocapnia during hyperthermic exercise reflects a reduction in the arterial CO2 tension (Pa(CO2)) set point. To test this hypothesis, five sheep were subjected to tracheal insufflations of CO2 or air (control) at 3-4 L min(-1) in 3 min bouts at 5 min intervals over 31 min of exercise. During exercise, rectal temperature and minute ventilation (V(E)) rose continuously while Pa(CO2) fell from 35.4+/-3.1 to 18.6+/ 2.9 Torr and 34.3+/-2.4 to 18.7+/-1.5 Torr in air and CO2 trials, respectively. Air insufflations did not affect V(E) or Pa(CO2). V(E) increased during CO2 insufflations via a shift to higher tidal volume and lower frequency. CO2 insufflations also increased Pa(CO2), although not above the pre-exercise level. Within 5 min after each CO2 insufflation, Pa(CO2) had decreased to match that following the equivalent air insufflation. These results are consistent with a reduced Pa(CO2) set point or an increased gain of the Pa(CO2) regulatory system during hyperthermic exercise. Either change in the control of Pa(CO2) could facilitate respiratory evaporative heat loss by mitigating homeostatic conflict. PMID- 15792597 TI - Physiological stress responses of two species of coral trout (Plectropomus leopardus and Plectropomus maculatus). AB - The physiological responses of two species of coral trout (Plectropomus maculatus and Plectropomus leopardus) to capture, shallow water and low salinity stressors were investigated. The responses of P. maculatus and P. leopardus to capture stress were characterised by rapid and transient increases in glucose, haemoglobin, haematocrit and lactate, as well as an equally dramatic but delayed increase in cortisol levels that persisted for at least 72 h. The magnitude and duration of the response to capture stress was very similar in both species. In contrast, the levels of cortisol, glucose, lactate, haemoglobin and haematocrit were generally elevated sooner and to higher levels in P. maculatus than in P. leopardus after exposure to shallow water stress. Coral trout exposed to reduced salinity showed minimal changes in cortisol, glucose, lactate, haemoglobin and haematocrit, but such changes were not characteristic of a non-specific response to stress. Thus, the physiological stress responses of coral trouts are species specific and dependent on the nature of the stressor. This observation probably reflects different cortical processes in the brains of P. maculatus and P. leopardus-a result that may be related to the differential variability of the respective environments in which the two species habit. PMID- 15792598 TI - A test of the thermal-stress and the cost-of-burrowing hypotheses among populations of the subterranean rodent Spalacopus cyanus. AB - Subterranean mammals show lower than-allometrically expected-basal metabolic rates (BMR), and several competing hypotheses were suggested to explain how physical microenvironmental conditions and underground life affect subterranean mammalian energetics. Two of these are the thermal-stress and the cost-of burrowing hypotheses. The thermal-stress hypothesis posits that a lower mass independent BMR reduces overheating in burrows where convective and evaporative heat loss is low, whereas the cost-of-burrowing hypothesis states that a lower mass-independent BMR may compensate for the extremely high energy expenditure of digging during foraging activity. In this paper we tested both hypotheses at an intraspecific level. We compared seven populations of the subterranean rodent Spalacopus cyanus or cururo from different geographic localities with contrasting habitat conditions. We measured BMR and digging metabolic rate (DMR) through open flow respirometry. Our results support neither the thermal-stress nor the cost-of burrowing hypothesis. Cururos from habitats with contrasting climatic and soil conditions exhibited similar BMR and DMR when measured under similar semi-natural conditions. It is possible that S. cyanus originated in Andean locations where it adapted to relatively hard soils. Later, when populations moved into coastal areas characterized by softer soils, they may have retained the original adaptation without further phenotypic changes. PMID- 15792599 TI - Metabolic responses of matrinxa (Brycon cephalus) to dietary protein level. AB - Effects of increasing dietary protein were studied in matrinxa, (Brycon cephalus), an omnivorous teleost from the Amazon Basin in Brazil. Four isocaloric diets were formulated to contain 20%, 27%, 34% and 41% of crude protein (CP). Plasma glucose levels were significantly increased while triacylglycerols were significantly reduced at 41% of CP. Free fatty acids were significantly reduced at each level of rising CP. Plasma amino acids and ammonium followed the dietary CP increase. Liver glycogen and amino acids were reduced; liver glucose and lactate were constant, and ammonium increased with the CP in diets. Muscle glycogen and pyruvate decreased, protein did not change, while lactate and free amino acids increased. Kidney glycogen proportionally rose with the increase of CP from 20-41%. Pyruvate and lactate augmented irregularly from 20-41% CP. A gluconeogenic profile was observed in the kidney; the liver worked as regulator of body glucose. Increase of dietary CP and decrease of carbohydrates pushed muscle and liver catabolism of fat and sugar stores to satisfy energetical demands. CP contents above 34% were not recommended for B. cephalus, if the carbohydrate does not compensate the metabolical demands. PMID- 15792600 TI - Inhibitory effects of the androgenic gland on ovarian development in the mud crab Scylla paramamosain. AB - Isolation and characterization of androgenic hormone in decapod crustaceans depend on an effective bioassay of its action. In the present study, the effect of androgenic gland on ovarian development in the mud crab Scylla paramamosain was investigated with a view to develop a bioassay for androgenic hormone. Ovarian regression with degeneration of oocytes occurred in some female crabs implanted with androgenic gland in vivo. In vitro incubation of ovarian tissues at secondary vitellogenesis in extract of androgenic gland resulted in a significant decrease in amino acid uptake by the tissues. We propose that this inhibitory effect could be established as an effective bioassay for the isolation of androgenic hormone in the mud crab. PMID- 15792601 TI - The acute hypoxic ventilatory response: testing the adaptive significance in human populations. AB - The acute Hypoxic Ventilatory Response (HVR) is an important component of human hypoxia tolerance, hence presumably physiological adaptation to high altitude. We measured the isocapnic HVR (L min(-1) %(-1)) in two genetically divergent low altitude southern African populations. The HVR does not differ between African Xhosas (X) and Caucasians (C) (X:-0.34+/-0.36; C:-0.42+/-0.33; P > 0.34), but breathing patterns do. Among all Xhosa subjects, size-independent tidal volume was smaller (X: 0.75+/-0.20; C: 1.11+/-0.32 L; P < 0.01), breathing frequency higher (X: 22.2+/-5.7; C: 14.3+/-4.2 breaths min(-1); P < 0.01) and hypoxic oxygen saturation lower than among Caucasians (X: 78.4+/-4.7%; C: 81.7+/-4.7%; P < 0.05). The results remained significant if subjects from Xhosa and Caucasian groups were matched for gender, body mass index and menstrual cycle phase in the case of females. The latter also employed distinct breathing patterns between populations in normoxia. High repeatability (intra-class correlation coefficient) of the HVR in both populations (0.77-0.87) demonstrates that one of the prerequisites for natural selection, consistent between-individual variation, is met. Finally, we explore possible relationships between inter-population genetic distances and HVR differences among Xhosa, European, Aymara Amerindians, Tibetan and Chinese populations. Inter-population differences in the HVR are not attributable to genetic distance (Mantel Z-test, P = 0.59). The results of this study add novel support for the hypothesis that differences in the HVR, should they be found between other human populations, may reflect adaptation to hypoxia rather than genetic divergence through time. PMID- 15792602 TI - The physiological and behavioral effects of carbon dioxide on Drosophila melanogaster larvae. AB - Adult and larval insects are rapidly anesthetized by carbon dioxide (CO2); however, the mechanisms have not been addressed. In this study, we use larval Drosophila to investigate the actions of CO2 to explain the behavioral effects of rapid immobilization and cardiac arrest with acute exposure to CO2. To determine if the central nervous system (CNS) is required, studies were performed with and without the CNS. The effects of low pH induced by exposure to CO2 were also examined. An acidic saline increases the heart rate in contrast to saline containing CO2. Synaptic transmission at the skeletal neuromuscular junction (NMJ) is blocked by CO2 but not by low pH. The site of action is postsynaptic by a decreased sensitivity to glutamate, the neurotransmitter at Drosophila NMJs. The CNS remains active in synaptic transmission when exposed to CO2 which is in contrast to the synapses at the NMJ. In summary, the effects of CO2 are directly mediated on the heart to stop it and at skeletal NMJs by a reduced sensitivity to glutamate, the released neurotransmitter, from the motor nerve terminals. The rapid behavioral and physiological effects cannot be accounted for by action on the CNS within the larvae nor by a pH effect indirectly induced by CO2. The glutamate receptors in the D. melanogaster preparation are similar in function to ionotropic glutamate receptors in vertebrates which could account for the observational phenomena of CO2 not yet explained mechanistically in vertebrates. PMID- 15792603 TI - Differential effects of temperature and maturity stage on hepatic estrogen receptor characteristics of Atlantic salmon. AB - In order to determine if elevated temperature during vitellogenesis had a detrimental effect on hepatic estrogen receptors of Atlantic salmon (Salmo salar), 3H-estradiol saturation binding analysis, using one- and two-site binding models, was carried out on extracts of hepatic cytosols from fish held at 14, 18 or 22 degrees C over the austral period of peak vitellogenesis (February to April). With one-site binding analysis, no temperature related difference in either receptor affinity (Kd) or number (Bmax) was found at each sampling point, but there was an apparent decrease in both affinity and number at each temperature over the period of the study. However, some analyses, notably at 22 degrees C during February, were best described using a two-site binding model. At this temperature and time, there was a clear separation of binding affinity into high and low components (Kd = 0.67+/-S.E. 0.05 and 20+/-S.E. 5.6 nM, respectively) (n = 4), which suggests that February was a critical time of temperature related hepatic sensitivity to estrogen. These results support those of other studies where we found that February was also a sensitive time with respect to temperature impairment of in vitro follicular estrogen synthesis, and the greatest period of in vivo temperature sensitivity. PMID- 15792604 TI - 4NQO oral carcinogenesis: animal models, molecular markers and future expectations. PMID- 15792605 TI - Idiopathic midline destructive disease: fact or fiction. AB - The differential diagnosis of a progressive destructive lesion of the midface and upper airway region includes both neoplastic and non-neoplastic entities; of these, the majority of cases prove to be either Wegener's granulomatosis or lymphoma. Historically, these sorts of necrotizing midfacial lesions were diagnosed clinically, and as a consequence a variety of overlapping categories of disease sprang up. As pathologic examination of biopsy material became both more widespread and (particularly in the last several years) more sophisticated, many lesions previously thought to be of mysterious origins have proven to be examples of lymphoma (in particular, sinonasal natural killer cell or T cell [NK/T] lymphomas). At present, the evaluation of a patient with a progressive destructive process involving the midface region should include imaging studies (to delineate the extent of disease) as well as biopsy (with sampling of lesional tissue for application of sophisticated testing--including immunohistochemical studies, flow cytometry, or molecular studies as necessary--to exclude the possibility of a NK/T cell lymphoma). There remain occasional patients whose necrotizing midfacial lesions continue to be difficult to classify despite the application of extensive testing; such patients are sometimes described as suffering from the nebulous entity of "idiopathic midline destructive disease". While it remains to be seen whether such patients will ultimately be assigned to other diagnostic groups (as, for example, occult toxic injuries--as in the case of cocaine abusers who are not forthcoming with regard to their drug usage), it seems likely that "idiopathic midline destructive disease" is a diagnostic term of questionable validity which should be used only with extreme reticence in modern practice. PMID- 15792606 TI - S100A2, a potential marker for early recurrence in early-stage oral cancer. AB - Early-stage oral cancer patients may have distinct clinical outcomes and respond differently to the same treatment. Up to now, there is still no individual marker to identify such patients with poor outcome. Down-regulation of a tumor suppressor gene, S100A2, in oral cancer cells was identified by mRNA profiling analysis then confirmed by RT-PCR and Southern blotting. The expression of nuclear S100A2 protein examined by immunohistochemistry was not significantly associated with any patient characteristic among the 70 early-stage oral squamous cell carcinoma (SCC) patients. Intriguingly, the loss of nuclear S100A2 positivity was significantly associated with shorter disease-free survival (p=0.019) while having no effect on the overall survival of these patients. Cox regression analysis with backward elimination identified S100A2 (p=0.006), tobacco smoking (p=0.013), and betel quid chewing (p=0.019) as independent predictors of disease-free survival. This is a first study to demonstrate that loss of nuclear S100A2 may serve as an independent prognostic marker for early stage oral cancer patients at high risk of recurrence. A more aggressive treatment modality and intensive follow-up may be recommended for the patients with reduced expression of S100A2 in tumor cell nuclei. PMID- 15792607 TI - Severe comorbidity negatively influences prognosis in patients with oral and oropharyngeal cancer after surgical treatment with microvascular reconstruction. AB - The aim of the study was to investigate the possible impact of comorbidity on survival of patients undergoing composite resection and microvascular reconstruction for oral/oropharyngeal cancer. Patient, tumour and treatment data were recorded. Comorbidity was graded by the Adult Comorbidity Evaluation (ACE 27) test. Survival and statistics were calculated. Comorbidity score ACE-27 grade 2 was present in 47% of patients, for ACE-27 grade 3 this was 13%. The median follow-up was 50 (3-87) months. Thirty-eight patients died, 32 developed a recurrence. Comorbidity score ACE-27 grade 3 turned out to be a clear predictor for overall survival (p<0.05). For ACE-27 grade 3 (n=13) 5-years survival was 29%, for ACE-27 grade 2 (n=87) this was 64%. No multivariate influences on the effects of comorbidity were found. Improved knowledge of the effect of comorbidity on survival may lead to better patient selection and counselling for major surgery and microvascular reconstruction. PMID- 15792608 TI - Genetic portrait of mild and severe lingual dysplasia. AB - Squamous cell carcinoma is the most frequent malignant tumor of the oral cavity and often arises from premalignant lesions. Traditional methods used by the pathologist are subjective and lack the sensitivity to predict accurately which precancers may progress with time. Therefore, it is important to search for markers that may identify progression of premalignant lesions. Microarray technology can be use with this aim. Here, we define the genetic expression profile of lingual dysplasia (DS) progression. By using cDNA microarray containing 19.2K clones and a baseline of 11 normal tissues, we compared 5 mild and 4 severe DS. We identified 270 genes differentially expressed in normal tissue vs. mild DS (i.e. 161 up- and 109 down-regulated) and 181 genes differentially expressed in mild vs. severe DS (i.e. 63 up- and 118 down regulated). The described genes cover a broad range of functional activities: (a) anti-oxidative, (b) DNA-repair, (c) inflammatory response, (d) cell adhesion/mobility, (e) extracellular matrix depolymerization, and (f) cell-cycle regulation. The data reported better define DS progression and can help in classifying premalignant lesions. PMID- 15792609 TI - TNFalpha played a role in induction of Akt and MAPK signals in ameloblastoma. AB - Tumour necrosis factor alpha (TNFalpha) is known crucial in inducing cell survival, proliferation, differentiation, and apoptosis. In the present study, we found that TNFalpha as well as its receptors, TNFR1 (TNF Receptor 1) and TNFR2, were clearly expressed in ameloblastoma tissues and AM-1 cells. By stimulation of TNFalpha in AM-1 cells, the phosphorylation of Akt (Ser473) and p44/42 mitogen activated protein kinase (MAPK) (Thr202/Tyr204) was markedly increased in TNFalpha concentration and time dependent manner. Pretreatment with U0126, mitogen-activated extracellular-regulated kinase (MEK) 1/2 inhibitor, prior to TNFalpha stimulation, specifically inhibited TNFalpha-induced phosphorylation of p44/42 MAPK (Thr202/Tyr204) in AM-1 cells. Meanwhile, pretreatment with LY294002, phosphatidylinositol-3-OH kinase (PI3K) inhibitor, could inhibit both TNFalpha induced phosphorylation of Akt (Ser473) and p44/42 MAPK (Thr202/Tyr204). These results suggested that TNFalpha is expressed in ameloblastoma and it can induce Akt and p44/42 MAPK activation through PI3K, which later might induce cell survival and proliferation in ameloblastoma. PMID- 15792610 TI - Ginkgo biloba extract (EGb 761) induces apoptosis by the activation of caspase-3 in oral cavity cancer cells. AB - We have investigated whether Ginkgo biloba extract (EGb 761) induces apoptosis of oral cavity cancer cells and attempted to characterize the apoptotic pathway activated by EGb 761. The inhibition of SCC 1483 oral cavity cancer cells proliferation was noted from 250 micro/ml of EGb 761. Apoptosis was observed after 24 h of incubation with 250 microg/ml EGb 761 and occurred in a time- and dose-dependent manner. Apoptosis was confirmed by DNA fragmentation and PARP cleavage. Co-treatment with the caspase inhibitor (z-VAD-fmk) inhibited apoptosis and PARP cleavage induced by EGb 761. Caspase-3 activity was upregulated by EGb 761 but reduced to the control level by co-treating with z-VAD-fmk. In summary, EGb 761 induces apoptosis of oral cavity cancer cells and caspase-3 is activated in this apoptosis. Therefore, EGb 761 may be considered as a possible chemopreventive agent against oral cavity cancer. PMID- 15792612 TI - The idiosyncratic relationship between diagnostic delay and stage of oral squamous cell carcinoma. AB - Approximately half of patients with oral cancer present with advanced lesions when five-year survival rates are as low as 20%, however, diagnostic delay has repeatedly been found to be unrelated to the stage of oral cancer at diagnosis. The aim of this study was to investigate why diagnostic delay is frequently unrelated to the stage of oral cancer at diagnosis. The odds ratio (OR) of early versus advanced oral squamous cell carcinoma was calculated for 245 consecutive patients with untreated oral squamous cell carcinoma. Being female (OR=0.40; 0.19 0.80; p<0.05) and married (OR=0.27; 0.11-0.66; p<0.01) was predictive of early stage disease. Being non-white (OR=5.42; 1.66-17.67; p<0.01) was predictive of advanced stage disease. There is no evidence that these patient demographics are associated with faster growing tumours, yet it is possible that those who are male, unmarried and non-white may not notice an oral lesion until it is advanced in stage. It is concluded that differences in tumour biology are not a likely explanation for the absence of a delay-stage relationship but instead, a proportion of tumours may be silent until advanced in stage. PMID- 15792611 TI - Early evaluation of the response to radiotherapy of patients with squamous cell carcinoma of the head and neck using 18FDG-PET. AB - The aim was to evaluate the efficacy of positron emission tomography (PET) with 2 [F-18]fluoro-2-deoxy-d-glucose (FDG) in early discrimination of response to definitive radiotherapy (RT) in patients with squamous cell carcinoma of the head and neck (SCCHN). Twenty-four patients who underwent FDG-PET scans before and after radiotherapy for nondisseminated SCCHN at the Asan Medical Center between August 2001 and September 2002 were prospectively evaluated. Initial FDG-PET scans were performed within 1 month before RT, and follow-up FDG-PET scans were performed 1 month after completion of RT. FDG-PET images were analyzed by standard uptake value (SUV). All patients were followed for more than 6 months. Pretreatment SUV ranged from 3.4 to 14.0 (median, 6.0), while posttreatment SUV ranged from ground level to 7.7 (median, 2.0). In evaluating residual tumors in these SCCHN patients, the overall sensitivity of FDG-PET was 100%, while its overall specificity was 87%. FDG-PET is effective in evaluating the response to radiation in patients with SCCHN. Timing the follow-up FDG-PET scan 1 month after completion of RT was not too rapid for evaluating the response to radiation. PMID- 15792613 TI - Development or existence of non-metastatic lymph nodes in the previously operated neck: a good prognostic sign for oromaxillary squamous cell carcinoma? AB - The aim of this study is to clarify the clinicopathological features of patients having non-metastatic lymph nodes in the previously operated neck for oromaxillary squamous cell carcinoma (OSCC). The clinicopathological factors of 9 such patients were comparatively analyzed with those of 78 other patients who underwent neck dissection (ND). The following observations were elicited from the data: (1) These 9 patients were alive and without OSCC for periods ranging from 1 year to 15 years and 9 months since their initial cancer treatments. (2) The interval between ND and clinical and/or imaging recognition of newly developed lymph nodes with suspicion of recurrence was significantly longer in these 9 patients. (3) The initially removed lymph nodes tended to be in a less advanced stage of disease. (4) Retrospectively, discrimination of non-metastatic lymph nodes from metastatic nodes was difficult in only one patient. (5) Each of the extirpated lymph nodes from 7 of the 9 patients showed a varying grade of reactive lymphadenitis on histopathology. In conclusion, the occurrence of benign hyperplastic lymph nodes in the previously operated neck region suggested favorable prognosis, though the immunological mechanism is not understood. PMID- 15792614 TI - Heterogeneity, histological features and DNA ploidy in oral carcinoma by image based analysis. AB - Oral squamous carcinomas appear heterogeneous on DNA ploidy analysis. However, this may be partly a result of sample dilution or the detection limit of techniques. The aim of this study was to determine whether oral squamous carcinomas are heterogeneous for ploidy status using image-based ploidy analysis and to determine whether ploidy status correlates with histological parameters. Multiple samples from 42 oral squamous carcinomas were analysed for DNA ploidy using an image-based system and scored for histological parameters. 22 were uniformly aneuploid, 1 uniformly tetraploid and 3 uniformly diploid. 16 appeared heterogeneous but only 8 appeared to be genuinely heterogeneous when minor ploidy histogram peaks were taken into account. Ploidy was closely related to nuclear pleomorphism but not differentiation. Sample variation, detection limits and diagnostic criteria account for much of the ploidy heterogeneity observed. Confident diagnosis of diploid status in an oral squamous cell carcinoma requires a minimum of 5 samples. PMID- 15792615 TI - Antiviral activity of Cidofovir on a naturally human papillomavirus-16 infected squamous cell carcinoma of the head and neck (SCCHN) cell line improves radiation sensitivity. AB - High risk HPV types 16 and 18 are associated with cervical cancer and squamous cell carcinoma of the head and neck (SCCHN). Cidofovir is an antiviral drug used to treat HPV-induced laryngeal papillomatosis and other viral infections, with initial reports suggesting activity in cervical carcinoma cells. We investigated the effects of Cidofovir on a naturally HPV-16-transformed SCCHN cell line (UPCI:SCC090), in comparison with a cervical carcinoma cell line (CasKi) of similar viral characteristics, to evaluate its therapeutic potential. HPV-16 gene transcription was only marginally reduced, and the antiviral and p53 restorative effects were modest in SCC90 cells. However, combination with irradiation enhanced the effects of Cidofovir treatment on these cells. Several days of treatment were required for this effect, which may limit its clinical applicability. Future therapies for HPV-associated tumors may include intralesional antiviral therapy in combination with radiation therapy, but optimization for clinical utility is needed. PMID- 15792616 TI - Cystic lymph node metastases of head and neck squamous cell carcinoma: pitfalls and controversies. AB - Lymph node metastases of squamous cell carcinomas in the neck can appear cystic. Without a clinically apparent primary tumour they can classically be considered to originate in a branchial cleft. We report two cases of cystic squamous cell carcinoma with histopathologic criteria of branchial cleft carcinoma. After a review of the literature, diagnostic strategies, histopathologic features, and therapeutic options for this very controversial clinical situation are discussed. We conclude by suggesting that Martin's criteria, largely used to differentiate between a cystic metastasis of a squamous cell carcinoma and a very theoretical branchial cleft carcinoma should be abandoned for lack of applicability. PMID- 15792618 TI - Age-dependent effects on biochemical variables and toxicity induced by cyclic peptide toxin microcystin-LR in mice. AB - Microcystins are naturally occurring hepatotoxins produced by certain strains of Microcystis aeruginosa and microcystin-LR is the most toxic among the 60 microcystin variants isolated so far. These toxins have been implicated in both human and livestock mortality. In the present study we evaluated the age dependent hepatotoxic effects of microcystin-LR (MC-LR) in mice after intraperitoneal and oral route of exposure. For acute toxicity studies by intraperitoneal route, 1 LD(50) dose of MC-LR (43.0 microg/kg) was administered to 6- to 36-week-old mice. Results showed that time to death in toxin treated animals decreased with age of mice. In comparison to control mice, treated animals of all age groups showed significant increases in liver body mass index and increases in serum enzymes (lactate dehydrogenase, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transpeptidase, sorbitol dehydrogenase). For acute oral toxicity studies, 1 LD(50) of microcystin-LR containing extracts (3.5 g of MCE/kg) was administered to 6- and 36-week-old mice. The effects on biochemical variables were similar to intraperitoneal route of exposure. Significant age-dependent effects that were observed in microcystin treated animals by intraperitoneal and oral routes of exposure include: time to death, hepatic lipid peroxidation, glutathione depletion and DNA fragmentation. The age-dependent effects observed in some of the biochemical variables may be due to difference in the amount of microcystin-LR up take and also the age dependent ability to detoxify the toxin in mice. PMID- 15792617 TI - Cisplatin as a radiosensitizer in the treatment of locally advanced head and neck cancer. PMID- 15792619 TI - Effect of mercuric chloride on electrical parameters and anion fluxes in the toad skin. AB - The amphibian skin, widely used for studying the transepithelial passage of electrolytes, exhibits anion pathways relatively specific for Cl(-). We studied the effect of HgCl(2), 1.0 x 10(-4) M on its electrical parameters and unidirectional anion fluxes. In the presence of Cl(-), the transepithelial conductance (G) of the isolated skin of the Bufo arenarum toad increased considerably following exposure to HgCl(2), whereas short-circuit current (SCC)- reflecting transepithelial Na(+) transport-underwent only slight stimulation. Following the blockade of Na(+) intake by amiloride, 1.0 x 10(-4) M, the removal of Cl(-) from the solution bathing the epidermal border of the skin brought about a decrease in G, and gave rise to a gradient-induced SCC (SCCg) consistent with transepithelial passage of Cl(-) along its gradient. Addition of mercaptoethanol, 5.0 x 10(-3) M to the bath containing Hg(2+) fully reversed these effects. The increase in G was accompanied by an increase in the unidirectional (epidermal to dermal) fluxes of (36)Cl(-) and (131)I(-), and a decrease in the passage of (99m)TcO(4)(-). These results show the effects of HgCl(2) to be similar to those of theophylline, although exhibiting a different selectivity. Our data suggest that anion passage following exposure to HgCl(2) is, like that stimulated by theophylline, predominantly if not exclusively transcellular, and does not involve a significant opening of the tight junctions. PMID- 15792620 TI - Hyper-osmoregulatory capacity of the Chinese mitten crab (Eriocheir sinensis) exposed to cadmium; acclimation during chronic exposure. AB - The aim of this study was to evaluate the effects of waterborne cadmium on hyper osmoregulatory capacity of the Chinese mitten crab Eriocheir sinensis acclimated to freshwater. For this purpose, crabs were submitted to acute (0.5 mg Cd L(-1) for 1, 2 or 3 days), chronic (10 or 50 microg Cd L(-1) for 30 days) or chronic, immediately followed by acute, exposure. While no effect was observed after 1 or 2 days, hemolymph osmolality, Na(+) and Cl(-) concentrations were significantly reduced after 3 days of acute exposure. Under this latter condition, the respiratory anterior gill ultrastructure, Na(+)/K(+)-ATPase and cytochrome c oxidase activities were significantly impaired. In contrast, the osmoregulatory posterior gill was unaffected for all treatments. As a consequence, we suggest that the observed hyper-osmoregulatory capacity impairment is the result of increased dissipative flow of ions and/or water through anterior gills. In contrast to acute exposure, chronic exposure did not induce any observable effect. However, crabs submitted to a known deleterious acute condition (0.5 mg Cd L(-1) for 3 days) directly after chronic exposure to 50 microg Cd L(-1) for 30 days showed normal hyper-osmoregulatory capacity with no change in gill Na(+)/K(+)-ATPase activity, and only little disturbance of anterior gill ultrastructure. These results demonstrate that a chronic cadmium exposure can induce acclimation mechanisms related to osmoregulation in this euryhaline decapod crustacean. PMID- 15792621 TI - Tissue-specific cadmium accumulation and metallothionein-like protein levels during acclimation process in the Chinese crab Eriocheir sinensis. AB - Aquatic organisms chronically exposed to cadmium can increase their resistance to a subsequent elevated exposure. In order to investigate mechanisms involved in acclimation process in the Chinese crab Eriocheir sinensis, we compared Cd level as well as metallothionein-like protein (MTLP) content in different tissues after direct acute exposure (i.e. 500 microg Cd L(-1) for 3 days), and after acute following chronic (i.e. 10 or 50 microg Cd L(-1) for 30 days) exposure. Cadmium accumulation occurred in the following order: anterior gill>hepatopancreas>posterior gill>carapace>hemolymph>muscle. As high concentrations as 188 microg Cd g(-1) w.w. were reported in anterior gills and seem to reach a saturation level. In these gills, the highest MTLP induction was observed after a direct acute exposure, for which a correlation with Cd content occurred. However, the Cd-binding potential by MTLPs was exceeded for any exposure condition. In hepatopancreas, the highest Cd level was reported for crabs acclimated during 30 days to 50 microg Cd L(-1) before challenging with an acute exposure. Moreover, we showed that MTLPs were induced during the acclimation process. In this organ, MTLPs are theoretically sufficient to bind all Cd. These results suggest that during a chronic exposure to 50 microg Cd L( 1), Chinese crabs acquire the capacity to hold more cadmium in hepatopancreas where it can be sequestrated by MTLPs. On the contrary, MTLP induction seems to be a rapid response to acute exposure in anterior gill, but is not sufficient to sequester all Cd. Other sequestration and/or detoxification mechanisms must take place in anterior gill to cope with high Cd levels. PMID- 15792622 TI - Gender differences in antioxidant capacity of rat tissues determined by 2,2' azinobis (3-ethylbenzothiazoline 6-sulfonate; ABTS) and ferric reducing antioxidant power (FRAP) assays. AB - Differences in susceptibility to oxidative stress between males and females have been postulated. Several methods have been developed to assess the total antioxidant capacity of human serum or plasma, but just recently some of them were employed for measurement of antioxidant capacity of tissues. In this study, we measured and compared antioxidant capacity of heart, kidney, liver and brain tissues of male and female rats. Antioxidant capacity was determined using 2,2' azinobis (3-ethylbenzothiazoline 6-sulfonate; ABTS) and ferric reducing antioxidant power (FRAP) assays. In the same samples, lipid peroxidation products of these tissues were analysed using thiobarbituric acid reactive substances (TBARS) assays. Antioxidant capacity of heart, kidney and liver tissues was higher in female than male rats for both FRAP and ABTS assays. We found positive correlation between FRAP and ABTS values for all tested tissues. FRAP and ABTS proved to be comparable, simple and quick methods for antioxidant capacity scanning in tissues. TBARS levels differed only for brain tissue, being higher in males. These results indicate stronger defense against oxidative damage in females for all observed tissues. These finding may account for the longer lifespan of females. PMID- 15792623 TI - Compensatory proteolytic responses to dietary proteinase inhibitors in the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae). AB - Increasing levels of inhibitors that target cysteine and/or serine proteinases were fed to Tribolium castaneum larvae, and the properties of digestive proteinases were compared in vitro. Cysteine proteinases were the major digestive proteinase class in control larvae, and serine proteinase activity was minor. Dietary serine proteinase inhibitors had minimal effects on either the developmental time or proteolytic activity of T. castaneum larvae. However, when larvae ingested cysteine proteinase inhibitors, there was a dramatic shift from primarily cysteine proteinases to serine proteinases in the proteinase profile of the midgut. Moreover, a combination of cysteine and serine proteinase inhibitors in the diet prevented this shift from cysteine proteinase-based digestion to serine proteinase-based digestion, and there was a corresponding substantial retardation in growth. These data suggest that the synergistic inhibitory effect of a combination of cysteine and serine proteinase inhibitors in the diet of T. castaneum larvae on midgut proteolytic activity and beetle developmental time is achieved through the prevention of the adaptive proteolytic response to overcome the activity of either type of inhibitor. PMID- 15792624 TI - Adaptations of the antioxidant system in erythrocytes of trained adult rats: impact of intermittent hypobaric-hypoxia at two altitudes. AB - We have investigated the effects of daily exposure to intermittent hypobaric hypoxia to two simulated altitudes (5700 m and 6300 m) in adult male rats that had been regularly swim trained in normoxia at sea level prior to exposures. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) along with the oxidative stress (OS) indices, malondialdehyde (MDA) and protein carbonyl content were measured in erythrocytes and their membranes. Hemoglobin increased in the trained animals exposed to 5700 m and in untrained rats exposed to 6300 m. Osmotic fragility in terms of hemolysis increased in altitude exposed animals. SOD increased in those exposed to 6300 m, while CAT increased in trained rats exposed to 5700 m and to 6300 m unlike in untrained rats where CAT increased only at 6300 m. GSH-Px showed varying degrees of elevation in all animals exposed to both altitudes. Erythrocyte membranes showed significant elevations in malondialdehyde (MDA) at 6300 m, while elevated protein carbonyls were noticeable at both altitudes in whole cells and membranes. These results suggest a positively associated elevation in protein oxidation with altitude in trained rats. At 5700 m, animals were less stressed, unlike at 6300 m, as seen from the magnitude of elevations in the OS indices and from the responses of the antioxidant enzymes. PMID- 15792625 TI - Molecular characterization and expression of the gene encoding aspartate aminotransferase from the Pacific oyster Crassostrea gigas exposed to environmental stressors. AB - A partial cDNA encoding cytosolic aspartate aminotransferase (AST) (EC 2.6.1.1) was isolated from a Crassostrea gigas digestive gland library. This sequence was used to design specific primers to amplify the AST genomic sequence. We obtained a complete gene, 5054 bp in length, encoding cytosolic AST and containing a 404 amino acid open reading frame. Phylogenetic analysis showed that C. gigas AST sequence constitutes a branch distinct from homologous sequences from other invertebrate groups. We also investigated AST mRNA expression in different tissues of oysters exposed to hydrocarbons, pesticides, hypoxia and hypo-salinity stress. The results showed that AST expression responds to hydrocarbon exposure, hypoxia and salinity stress, but not to pesticide exposure in an organ and time specific manner. Use of AST as a potential molecular biomarker for monitoring of disturbed ecosystems is discussed. PMID- 15792626 TI - Exposure of tilapia pituitary cells to saponins: insight into their mechanism of action. AB - Cell permeation and durable effects of triterpenoidal saponin preparations from soybean (SbS), Quillaja saponaria Molina (QsS) and Gypsophila paniculata (GypS), were studied. A concentration-dependent change in hemolysis rates was observed when cells were incubated with QsS or GypS, but not with SbS. Dose dependence was also observed for the leakage of lactate dehydrogenase (LDH; MW 142,000) and of Luteinizing Hormone (LH; MW 35,000) from tilapia pituitary dispersed cells. Exposure of pituitary fragments to a combination of GnRH and GypS or QsS, resulted in a significantly high release of LH. GypS were shown to be more potent in inducing hemolysis of human RBC's and LH release from tilapia pituitary fragments. Interestingly, tilapia pituitary fragments treated with QsS were able to secrete LH in a characteristic manner, in response to a second Gonadotropin Releasing Hormone (GnRH) pulse, while fragments exposed to GypS did not respond to the second hormone pulse. The rapid recovery of pituitary fragments after the removal of QsS, may suggest a rearrangement of membranes rather than pore formation as the mechanism of action of QsS. Understanding the structural features underlying the reversible rearrangement of membranes and the lack of hemolysing activity by specific saponins may lead to the development of novel bioactive drugs. PMID- 15792627 TI - Effects of nonylphenol on hepatic testosterone metabolism and the expression of acute phase proteins in winter flounder (Pleuronectes americanus): comparison to the effects of Saint John's Wort. AB - 4-Nonylphenol (4-NP), a major by-product of alkylphenol ethoxylates, is used in several industries and as a consequence is quite common in rivers, estuaries and other aquatic environments that receive sewage discharges or are near offshore oil platforms. 4-NP is an environmental estrogen that also binds human and rodent Pregnane X-receptor (PXR), the orphan nuclear receptor that controls the expression of several detoxication genes in mammals, including several CYP3A and CYP2B family members. These P450s preferentially hydroxylate testosterone in the 6beta- and 16beta-positions, respectively. In this study, the effects of 4-NP on testosterone metabolism and hepatic CYP3A induction were compared to the effects of St. John's Wort (SJW), a well established mammalian PXR agonist, in winter flounder. Male winter flounder (Pleuronectes americanus) were injected with 100 mg/kg/day 4-NP or 500 mg/kg/day SJW or both (S and N) every 24 h. Forty-eight hours after the initial injections, flounder were euthanized. Western blots and testosterone 6beta-hydroxylation indicated that CYP3A was increased 50% by 4-NP, but was not affected by SJW. Testosterone 16beta-hydroxylase activity was also significantly increased in flounder treated with 4-NP (2.8 x), but not with SJW. This is not consistent with our hypothesis that both SJW and 4-NP would induce CYP3A. Subtractive hybridization was performed between control and 4-NP treated hepatic mRNA samples to isolate differentially expressed genes. Subtractive hybridization indicated that several acute phase proteins were altered by 4-NP. Quantitative real-time PCR (Q-PCR) confirmed 4-NP altered the expression of complement components C8b, cathepsin L, C-type lectin domain, FK506 binding protein 2 precursor (FKBP2) and an EST (expressed sequence tag). SJW and 4-NP treated flounder demonstrated similar induction profiles for the EST, cathepsin L and FKBP2, suggesting that SJW was at a sufficient dose to alter gene expression but not induce P450s. In conclusion, testosterone hydroxylase activity and Western blots indicate that SJW did not activate detoxication pathways in a similar manner to 4-NP. PMID- 15792628 TI - Hematology patterns of migrating European eels and the role of EVEX virus. AB - We show that European eels infected with the rhabdovirus EVEX (Eel Virus European X) virus, developed hemorrhage and anemia during simulated migration in large swim tunnels, and died after 1000-1500 km. In contrast, virus-negative animals swam 5500 km, the estimated distance to the spawning ground of the European eel in the Sargasso Sea. Virus-positive eels showed a decline in hematocrit, which was related to the swim distance. Virus-negative eels showed a slightly increased hematocrit. Observed changes in plasma lactate dehydrogenase (LDH), total protein and aspartate aminotransferase (AAT) are indicative of a serious viral infection. Based on these observations, we conclude that eel virus infections may adversely affect the spawning migration of eels, and could be a contributing factor to the worldwide decline of eel. PMID- 15792629 TI - Metabolic changes in Japanese medaka (Oryzias latipes) during embryogenesis and hypoxia as determined by in vivo 31P NMR. AB - In vivo (31)P nuclear magnetic resonance spectroscopy (NMR) was used to determine phosphometabolite changes in medaka (Oryzias latipes) during embryogenesis and hypoxia. NMR data were acquired using a flow-through NMR tube perfusion system designed to both deliver oxygenated water to embryos and accommodate a hypoxic challenge. Measurements of embryogenesis at 12- and 24-h intervals throughout 8 days of development (n = 3 per time point, 900 embryos per replicate) and during acute hypoxia (n = 6, 900 embryos at Iwamatsu stage 37 per replicate) were performed via NMR, and replicate samples (n = 4, 250 embryos each) were flash frozen for HPLC analysis. The hypoxic challenge experiment consisted of data acquisition with recirculating water (pre-hypoxic control period; 1 h), without recirculating water (hypoxic challenge; 1 h), then again with recirculating water (recovery period; 1.3 h). Concentrations of ATP, phosphocreatine (PCr), orthophosphate (P(i)), phosphomonoesters (PME), phosphodiesters (PDE), and intracellular pH (pH(i)) were determined by NMR, and ATP, ADP, AMP, GTP, GDP, and PCr were also determined via HPLC. During embryogenesis, [ATP] and [PCr] as determined by HPLC increased from 1-day post fertilization (DPF) levels of 0.93+/ 0.08 and 2.48+/-0.21 micromol/mg (dry tissue), respectively, to 7.24+/-0.77 and 15.66+/-1.08 micromol/mg, respectively, by day 8. [ATP] and [PCr] measured by both NMR and HPLC fluctuated over 1-3 DPF, then increased significantly (p<0.05) over 3-8 DPF, while [PME] and [PDE] decreased (p<0.05) throughout embryogenesis. NMR and HPLC measurements revealed 1-3, 4-5, and 6-8 DPF as periods of embryogenesis significantly different from each other (p<0.05), and representing important transitions in metabolism and growth. During hypoxic challenge, [ATP] and [PCr] declined (p<0.05), [PME] and [PDE] decreased slightly, and [P(i)] increased (p<0.05). All phosphometabolites returned to pre-hypoxia concentrations during recovery. The pH(i) decreased (p<0.05) from 7.10+/-0.03 to 6.94+/-0.03 as a result of hypoxia, and failed to return to pre-hypoxic levels within the 1.3-h recovery phase. Results demonstrate the utility of in vivo (31)P NMR to detect significant alterations in phosphorylated nucleotides and phosphometabolites at specific developmental stages during medaka development and that late-stage medaka utilize PCr to generate ATP under hypoxic conditions. PMID- 15792630 TI - Hydroxyl radical production and oxidative damage induced by cadmium and naphthalene in liver of Carassius auratus. AB - Freshwater goldfish (Carassius auratus) were exposed to cadmium (Cd) from 0 to 5 mg/L, and naphthalene (NAP) from 0 to 50 mg/L. Twenty-four hours after the exposure, reactive oxygen species (ROS) was trapped by phenyl-tert-butyl nitrone and detected by electron paramagnetic resonance (EPR). Protein carbonyl (PCO) and lipid peroxidation (LPO) content were determined. The activities of superoxide dismutase (SOD) and catalase (CAT) were also measured. The EPR spectra signals were characterized by prominent six-line spectra, which were defined as hydroxyl radical ((.)OH). As compared to the control group, Cd and NAP significantly induced (.)OH production marked by the intensity of the prominent spectra at higher concentrations. Both xenobiotics also increased LPO content and PCO content, depending on the concentrations. Either LPO or PCO content showed significant relation with (.)OH production. Cd increased the activity of SOD and decreased that of CAT at 5 mg/L, and NAP increased the activities of SOD and CAT at 5 mg/L. The results clearly indicated that these two structurally different non-redox cycling xenobiotics could induce (.)OH generation and result in oxidative damage in liver of C. auratus, and these effects were concentration dependent. PMID- 15792631 TI - Distinct effects of 4-nonylphenol and estrogen-17 beta on expression of estrogen receptor alpha gene in smolting sockeye salmon. AB - Xenoestrogens such as 4-nonylphenol (4-NP) have been shown to affect the parr smolt transformation, but their mechanisms of action are not known. We therefore examined effects of 4-NP and estradiol-17beta (E2) on expression of estrogen receptor (ER) alpha gene in the liver, gill, pituitary and brain of sockeye salmon to elucidate molecular mechanisms of 4-NP and E2 and developmental differences in response during smolting. Fish were treated twice within a week with 4-NP (15 and 150 mg/kg BW), E2 (2 mg/kg BW) or only vehicle at three stages of smolting, pre-smolting in March, early smolting in April and late smolting in May. The absolute amounts of ERalpha mRNA were determined by real-time PCR. The basal amounts of ERalpha mRNA peaked in April in the liver, gill and pituitary. In March, E2 extensively increased the amounts in the liver, while 4-NP had no effects at this stage. In contrast, 4-NP (but not E2) decreased liver ERalpha mRNA in April. 4-NP also decreased the amount of ERalpha mRNA in the gill in April. In the pituitary, 4-NP increased ERalpha mRNA in March but decreased it in May. There were no significant effects in the brain. Changes in basal ERalpha mRNA observed in this study indicate that estrogen responsiveness of tissues may change during salmon smolting. Furthermore, 4-NP and E2 have different effects on expression of ERalpha gene in the liver and gill during smolting, and the response is dependent on smolt stage. PMID- 15792632 TI - Responses of hybrid striped bass to waterborne and dietary copper in freshwater and saltwater. AB - Mechanisms of copper toxicity and consequences of exposure vary due to uptake route and ionoregulatory status. The goal of this research was to develop a model fish system to assess the influence of different Cu exposure routes (waterborne or dietary) on bioavailability, uptake, and effects in hybrid striped bass (Morone chrysops x Morone saxatilis) acclimated to fresh- or saltwater. Initially, hybrid striped bass were exposed to dietary Cu concentrations of 571, 785, and 1013 mug Cu/g, along with a control (approximately 5 microg Cu/g), for 14 days in saltwater. Intestinal and liver Cu accumulated in a dose-dependent manner in fish exposed to increasing levels of dietary Cu. Chronic (42 days) experiments were then conducted to determine sub-lethal effects of aqueous, dietary, and combined aqueous and dietary Cu exposures to both freshwater- and saltwater-acclimated hybrid striped bass. Growth and Cu accumulation in the gill, intestine, and liver were measured. Although no significant effects were observed in fish exposed to waterborne Cu, those exposed through the diet accumulated significant liver and intestinal Cu but showed no significant change in growth. Overall, these results suggest that at the levels tested, exposure to elevated waterborne Cu did not cause significant long-term tissue Cu accumulation, whereas dietary Cu exposure caused significant liver and intestinal Cu accumulation in hybrid striped bass which was comparable in both freshwater and saltwater (15 g/L). PMID- 15792633 TI - Carbonic anhydrase metabolism is a key factor in the toxicity of CO2 and COS but not CS2 toward the flour beetle Tribolium castaneum [Coleoptera: Tenebrionidae]. AB - The analogues carbon dioxide (CO(2)), carbonyl sulfide (COS) and carbon disulfide (CS(2)) have been useful as substrate probes for enzyme activities. Here we explored the affinity of the enzyme carbonic anhydrase for its natural substrate CO(2), as well as COS and CS(2) (1) by in vitro kinetic metabolism studies using pure enzyme and (2) through mortality bioassay of insects exposed to toxic levels of each of the gases during carbonic anhydrase inhibition. Hydrolysis of COS to form hydrogen sulfide was catalysed rapidly showing parameters K(m) 1.86 mM and K(cat) 41 s(-1) at 25 degrees C; however, the specificity constant (K(cat)/K(m)) was 4000-fold lower than the reported value for carbonic anhydrase-catalysed hydration of CO(2). Carbonic anhydrase-mediated CS(2) metabolism was a further 65,000-fold lower than COS. Both results demonstrate the deactivating effect toward the enzyme of sulfur substitution for oxygen in the molecule. We also investigated the role of carbonic anhydrases in CO(2), COS and CS(2) toxicity using a specific inhibitor, acetazolamide, administered to Tribolium castaneum (Herbst) larvae via the diet. CO(2) toxicity was greatly enhanced by up to seven fold in acetazolamide-treated larvae indicating that carbonic anhydrases are a key protective enzyme in elevated CO(2) concentrations. Conversely, mortality was reduced by up to 12-fold in acetazolamide-treated larvae exposed to COS due to reduced formation of toxic hydrogen sulfide. CS(2) toxicity was unaffected by acetazolamide. These results show that carbonic anhydrase has a key role in toxicity of the substrates CO(2) and COS but not CS(2), despite minor differences in chemical formulae. PMID- 15792635 TI - Anti-opsonic properties of staphylokinase. AB - Recently we described a novel bacteriophage-encoded pathogenicity island in Staphylococcus aureus that harbors a number of virulence factors that are all involved in the evasion of innate immunity. Here we describe a mechanism by which staphylokinase (SAK), frequently present on this pathogenicity island, interferes with innate immune defenses: SAK is anti-opsonic. By activating human plasminogen (PLG) into plasmin (PL) at the bacterial surface, it creates bacterium-bound serine protease activity that leads to degradation of two major opsonins: human immunoglobulin G (IgG) and human C3b. Incubation of opsonized bacteria with PLG and SAK resulted in removal of anti-staphylococcal IgGs and C3b from the bacterial surface. In phagocytosis assays this proved to be a very efficient mechanism to reduce the opsonic activity of human IgG and serum. The fact that SAK activates human PLG at the bacterial surface and removes IgG as well as C3b makes this protein a unique anti-opsonic molecule. PMID- 15792634 TI - Gene structure and promoter analysis of the human GJB6 gene encoding connexin 30. AB - Connexins (Cx) are the protein subunits of gap junctions, which play an important role in cell-to-cell communication. We characterized the genomic structure of the human GJB6 gene, encoding connexin 30 (C x 30), and showed that it differs from most connexin-encoding genes. GJB6 presents six different exons, some of which can be alternatively spliced. We also mapped a basal promoter sequence active in a human keratinocyte cell line which responds to the activation of the EGF receptor. One of the non-encoding exons of GJB6, which has been described in brain C x 30 cDNA, was not found in cDNA obtained from human keratinocytes, suggesting tissue-specific splicing. PMID- 15792636 TI - Interferons (IFNs) are key cytokines in both innate and adaptive antiviral immune responses--and viruses counteract IFN action. AB - Interferons (IFNs) are transcriptionally regulated cytokines and key players in the innate antiviral immune response. Upon recognition of a virus, or its molecular patterns, by the Toll-like receptors of dendritic cells (DCs), high levels of IFNs are expressed by these cells. This in turn stimulates DC maturation and the subsequent expression of proinflammatory cytokines and costimulatory molecules, leading to the transition to an adaptive antiviral immunity. Conversely, viruses have developed diverse strategies to counterattack host defenses in order to generate their progenies. PMID- 15792637 TI - A nose-only apparatus for airborne delivery of Mycobacterium tuberculosis to mice: calibration of biological parameters. AB - Mycobacterium tuberculosis is the main cause of tuberculosis and is still a public health concern worldwide. This mycobacterium is transmitted through aerosols from human beings suffering from pulmonary tuberculosis to susceptible persons. To study this natural route of infection, we designed a new nose-only aerosol apparatus--system of aerosolisation of microorganisms (SAM)--in a carefully designed biohazard facility. For safety reasons, Mycobacterium smegmatis was first used to calibrate several parameters, such as inoculum density, atmospheric conditions (i.e. hygrometry) and particle size distribution. We present evidence that our apparatus is totally adapted to airborne delivery; the particle size of generated aerosol ranges from 1 to 7 microm, which is ideal for an infection by inhalation. We found that 99% of generated particles (<7 microm) could be retained by the respiratory tract, and among these particles, 62 79% (<3.3 microm) were able to reach pulmonary compartments. The next step was to simultaneously challenge 48 mice with M. tuberculosis in a highly reproducible way. We showed that a moderate dose (4 log10 colony-forming units (CFU) per mice) of M. tuberculosis was capable of causing progressive lung pathology and death in mice 30 days post-aerosolisation. Therefore, our apparatus, once calibrated, is easy to handle, safe, and can be used with any pathogen, which is spread by aerosol. PMID- 15792638 TI - Presence of fibronectin-binding protein gene prtF2 in invasive group A streptococci in tropical Australia is associated with increased internalisation efficiency. AB - The fibronectin-binding proteins (FnBPs) PrtF1 and PrtF2 are considered to be major group A streptococcal virulence factors, mediating adherence to and internalisation of host cells. The present study investigated an association between the presence of prtF1 and prtF2 genes and internalisation efficiency in group A streptococci (GAS) isolated from patients with invasive disease. Of the 80 isolates tested, 58 (73%) had prtF1 and 71 (89%) possessed prtF2. Three isolates (4%) had neither gene, seven (9%) had prtF1 only, 19 (24%) had prtF2 only and 51 isolates (64%) had both prtF1 and prtF2. prtF2-positive isolates internalised up to three times more efficiently than isolates that had prtF1 alone (P<0.001), and 1.5-fold better than isolates that had neither gene. No significant association was found between internalisation efficiency and presence of the prtF1 gene. Analysis of the fibronectin-binding repeat domain (FBRD) of prtF2 revealed that this gene can contain 2, 3, 4 or 5 repeat regions and that five repeat regions conferred very high internalisation efficiency in invasive GAS isolates. PMID- 15792639 TI - Acute hepatitis E of a man who consumed wild boar meat prior to the onset of illness in Nagasaki, Japan. AB - Zoonotic infection of hepatitis E virus (HEV) has been suggested. To date, pigs, deer, and wild boar have been implicated as reservoirs of HEV in Japan. However, it is not known to what extent zoonotic transmission of HEV play roles of causing HEV transmission. In the present report, we describe a case of acute hepatitis E in which a transmission of HEV by a zoonotic transmission is strongly suggested. The patient had eaten grilled wild boar meat 59 days prior to onset of acute hepatitis. Although the meat was not stored, one of the two people who ate boar meat with the patient at the same time showed high levels of HEV-IgM and -IgG and normal levels of liver enzymes, suggesting a subclinical infection of HEV. Accumulating evidence suggests that eating wild boars is associated with a high risk of acquiring hepatitis E infection. PMID- 15792640 TI - Clinical comparison of branched-chain amino acid (l-Leucine, l-Isoleucine, l Valine) granules and oral nutrition for hepatic insufficiency in patients with decompensated liver cirrhosis (LIV-EN study). AB - This multicenter study compared the effects of branched-chain amino acid granules (Livact((R)) Granules, LIV) and an enteral nutrient for chronic hepatic failure (Aminoleban((R)) EN, EN) on serum albumin in patients with decompensated liver cirrhosis. This study enrolled "patients with decompensated liver cirrhosis associated with hepatic encephalopathy who were suffering from hypoalbuminemia in spite of adequate food intake," a condition for which both drugs are indicated. Enrolled patients were randomized to the two groups according to the central registration method. This study continued for 24 weeks. Selected foods were supplied to each patient in principle so that caloric and protein intakes were standardized between the two groups. A total of 281 patients were enrolled. LIV was not inferior to EN concerning the primary efficacy endpoint changes in serum albumin. PMID- 15792641 TI - Risk factors of delay in restoration of hepatic reserve capacity and local recurrence after radiofrequency ablation therapy for hepatocellular carcinoma (HCC). AB - BACKGROUND: : Effects of radiofrequency ablation (RFA) on hepatic reserve capacity have not been evaluated thoroughly thus far. The aim of our study is to evaluate the factors that influence to hepatic reserve capacity and local recurrence after RFA. PATIENTS AND METHODS: : We studied a total of 243 patients (310 nodules). The study parameters included rates of local recurrence after undergoing RFA as well as factors for delaying recovery of post-RFA albumin levels (age, gender, the presence and absence of anti-HCV antibody, platelet count, GPT level, prothrombin time (PT), Child-Pugh grade, pre-RFA albumin level, alpha-fetoprotein (AFP) level, the number of nodules, tumor size in diameter, hepatic blood control and the coagulation volume after undergoing RFA. RESULTS: : Rates of local recurrence were 6.5%, 10.4% and 12.0% at 1, 2 and 3 years after undergoing RFA, respectively. In the hepatic reserve capacity studies, it took 1.9 and 5.8 months for 50% and 80% of cases, respectively, to restore serum albumin levels to pre-treatment levels after undergoing RFA. There were significant differences in the time of delay in restoration of serum albumin levels with regard to two factors: patients' age >/=60 years (p=0.0351) and tumor size over 3.5cm in diameter (p<0.001). CONCLUSION: : We cosidered that RFA is a safe and efficacious modality, but thorough attention was necessary for such patients with tumor size over 3.5cm in diameter before undergoing RFA, especially in elderly patients. PMID- 15792642 TI - The World Health Organization initiative on implantation research. AB - Recognizing that the currently available contraceptive options represent a limited choice for women, contraceptive research and development programs have identified the process of implantation as a promising area for investigation. Contraceptive methods that prevent implantation and can be taken on a once-a month basis appear to be an attractive option for many women in a variety of settings. A recently completed 5-year, international, collaborative research programme - conducted by the World Health Organization with financial support from the Rockefeller Foundation - has identified a number of promising leads for once-a-month method development and the more promising of these leads are being actively followed up in collaboration with the pharmaceutical industry. PMID- 15792643 TI - Intraovarian actions of anti-angiogenic agents disrupt periovulatory events during the menstrual cycle in monkeys. AB - To determine if anti-angiogenic agents disrupt primate ovarian function, vehicle or a general angiostatic compound (TNP-470), specific antagonists of vascular endothelial growth factor (soluble VEGF receptor-1, sVEGFR-1; anti-VEGF monoclonal antibody, VEGF Ab) and/or an angiopoietin antagonist (Ang-2) were administered to rhesus monkeys: (1) locally via injection into the preovulatory follicle at midcycle or the developing corpus luteum at the midluteal phase; or (2) systemically via subcutaneous injection in the early follicular phase or at midcycle during the natural menstrual cycle. Compared to controls, intrafollicular injection of TNP-470 or sVEGFR-1 decreased circulating progesterone (P) levels in the subsequent luteal phase. Treatment with sVEGFR-1, but not TNP-470, also decreased the incidence of ovulation. Intrafollicular injection of Ang-2 also prevented ovulation, as well as any functional luteal phase. In the absence of elevated P, serum estradiol levels rose to peak levels 11-12 days post-Ang-2 treatment, at which time another large antral follicle was observed on the contralateral (noninjected) ovary. Intraluteal and systemic injection of VEGF antagonists alone or with Ang-2 had minimal effects. Thus, anti angiogenic factors can act locally in the primate follicle to disrupt the gametogenic (oocyte release) and endocrine (steroid) functions of the ovary. However, further studies are needed to optimize delivery of angiogenic agents before they can be meaningfully evaluated as possible contraceptive agents. PMID- 15792644 TI - Involvement of molecules related to angiogenesis, proteolysis and apoptosis in implantation in rhesus monkey and mouse. AB - We have established the well-defined cycling, pseudo-pregnant and pregnant rhesus monkey models, and used these to analyze expression of the common molecules specifically related to angiogenesis, apoptosis or proteolysis, such as vascular endothelial growth factor (VEGF) and its receptors KDR, flt-1, flt-4 and flk-1, basic fibroblast growth factor (bFGF) and its receptors Flg, transforming growth factor-alpha and beta1 (TGF-a/beta1), and TGF-beta1 receptor type I (TbetaR-I) and type II (TbetaR-II), as well as steroidogenic acute regulatory protein (StAR), tissue type plasminogen activator/urokinase plasminogen activator/plasminogen activator inhibitor type 1 (tPA/uPA/PAI-1) and matrix matalloproteinase type 1, -3/tissue inhibitor matalloproteinase type 1, -2, -3 (MMP-1, -3/TIMP-1, -2, -3), Fas/FasL, BcL-2/Bax, in the corpus luteum (CL), in the functional layer of the endometrium and in the materno-fetal boundary of the implantation site. We have demonstrated that: expression of these molecules in the monkey CL, endometrium and materno-fetal boundary of the implantation site is correlated well with CL functional and vascular development and with the processes involved in the establishment of the implantation window as well as with the early stages of placentation. A coordinated increase in tPA and its inhibitor PAI-1 expression in the monkey and rat CL may be instrumental in initiating luteal regression in both species, and correlated well with the timing of the closure of the implantation window, whereas high uPA activity in the CL is important for the early formation of the CL and for maintaining its function which is closely correlated to the period of establishment of the implantation window. Apoptosis, proteolysis and angiogenesis occur in the CL and in the endometrium during the time of establishment of the implantation window, as well as in the materno-fetal boundary of the implantation site at the early stages of placentation. It seems that these processes occur in these tissues in a coordinated and time- and cell-dependent manner, and are reliant on each other. Based on these observations, we have designed experiments to test the actions of some related available compounds on mouse implantation, used alone or in combination. The preliminary data showed that the compounds which could effectively affect apoptosis, angiogenesis or proteolysis in the implantation site were capable of effectively inhibiting implantation by acting on the endometrium and/or on the CL. Furthermore, the combined use of these compounds produced an obvious additive effect on inhibiting implantation. This finding suggested this may be a good approach for developing an anti-implantation agent. PMID- 15792645 TI - Novel antiangiogenic agents for use in contraception. AB - Angiogenesis and vascular development are fundamental to the development of a receptive endometrium that permits implantation. The underlying hypothesis of this project is that implantation in primates and in humans is dependent on vascular remodeling in the endometrium and that the identification of agents that can disrupt this process prior to embryo attachment will lead to the development of new post coital contraceptives. To identify suitable targets for postcoital contraception, we studied the expression in endometrium of the vascular endothelial growth factor (VEGF) and angiopoietin families of angiogenic regulators. We produced a neutralizing antibody to VEGF-A, and this was shown to inhibit implantation in rhesus monkeys, apparently through direct antagonism of the action of VEGF-A in the endometrium. This demonstrated 'proof of principle' that agents antagonizing molecules that regulate angiogenesis can be developed as contraceptive agents. A second objective was to identify new contraceptive targets. We have developed microarrays to compare receptive endometrium with endometrium-rendered nonreceptive by a number of experimental strategies. We have identified over 100 RNA transcripts that are acutely regulated by administration of the antiprogestin RU486 to women, and 20 transcripts altered by antagonizing the action of VEGF-A in endometrium. These transcripts represent new potential targets for development of novel postcoital contraceptives. PMID- 15792646 TI - Identification of novel endometrial targets for contraception. AB - Successful embryo implantation is a critical step in establishing pregnancy and requires appropriate preparation of the endometrium to provide a transient state of "uterine receptivity." The most essential of the molecular events determining receptivity may therefore provide potential targets for postcoital contraception. Using the mouse as a model, we identified molecules specifically regulated in the endometrium at very early implantation: these were monoclonal nonspecific suppressor factor beta (MNSFbeta), splicing factor SC35, a novel protease of the HtrA family, termed HtrA3, calcium-binding protein (CaBP)-d9k (calbindin d9k) and proprotein convertase 6 (PC6). All of these molecules were also expressed in human endometrium, with the exception of CaBP-d9k, which was represented by the functionally similar CaBP-d28k. Appropriate spatial and temporal expressions of mRNA and protein were demonstrated for all five candidate molecules in mouse and primate (human and rhesus monkey) endometrium during the menstrual cycle and early pregnancy. Functional studies in mice established that blocking production of the CaBPs and PC6 within the endometrium completely prevented implantation and thus provided proof of principle that these molecules are potential contraceptive targets. PMID- 15792647 TI - Evaluation of fusogenic trophoblast surface epitopes as targets for immune contraception. AB - Syncytial trophoblast fusion is an essential step in the process of implantation. This project is aimed at the immunological inhibition of syncytial trophoblast fusion as a novel approach to contraception. Fusion-inhibiting recombinant antibodies were generated and used together with autoantibodies from patients with repetitive in vitro fertilization (IVF) failure that were shown to inhibit syncytial fusion and are expected to inhibit implantation, to generate anti idiotypic peptides. These peptides mimic trophoblast epitopes essential for syncytial fusion and are, therefore, considered specific immunogens for the generation of antibodies that will inhibit implantation. To verify their physiological role in humans, 300 anti-idiotypic peptides were tested for their binding capacity to patient autoantibodies associated with repetitive IVF failure, habitual abortion and preeclampsia. Of these, only three peptides were found to selectively bind to autoantibodies of patients with repetitive IVF failure and were considered safe and efficient enough for evaluation in preclinical and clinical studies required for the development of immune contraceptives. When used as immunogens, these peptides are expected to elicit an antibody response inhibiting syncytial fusion and thus implantation. Furthermore, the action of these antibodies needs to be restricted to the stage of syncytium formation at the time of implantation so as not to cause complications of pregnancy in those cases where they fail to have a contraceptive effect. To exclude potential side effects on other systems, toxicological experiments in animals are in progress. PMID- 15792648 TI - Target-oriented anti-implantation approaches for pregnancy interception: experiences in the rhesus monkey model. AB - Blastocyst implantation is a critical process in the establishment of pregnancy in eutherian mammals and requires a harmonious symbiosis between the developing conceptus and the differentiating maternal uterus. A better understanding of this symbiotic relationship will provide novel approaches and interventions for realizing anti-implantation strategies for effective fertility regulation and reproductive health care management. We have been using the rhesus monkey (Macaca mulatta) as a nonhuman primate model to this end. In the present study, the process of progesterone-mediated regulation of endometrial receptivity for blastocyst implantation has been targeted by the use of mifepristone as an emergency contraceptive agent. Furthermore, based on cell-specific, temporal and spatial distribution of vasotropic cytokines and mediators in the "receptive" and periimplantation periods, the pregnancy interceptive potentials of (a) monoclonal antibody (MAb) to leukemia inhibitory factor (LIF); (b) inhibitors of nitric oxide synthase [e.g., N6-nitro-l-arginine (l-NAME) and aminoguanidine]; and (c) MAb to vascular endothelial growth factor (VEGF) were examined. LIF is a progesterone-responsive pleiotropic cytokine that functions as a proinflammatory cytokine, together with interleukins 1 and 6, during the process of implantation placentation in primates, and its immunoneutralization with MAb resulted in inhibition (p<.04) of pregnancy establishment in the rhesus monkey. However, timed administration of l-NAME or aminoguanidine failed to inhibit blastocyst implantation in a significant manner. Also, no synergistic antinidatory action of antiprogestin combined with l-NAME was detected in the rhesus monkey. The application of MAb to VEGF during the periimplantation period, on the other hand, led to significant (p<.04) prevention of pregnancy without influencing steroid hormone levels in the circulation. Our data lend support to the hypothesis that VEGF is essential for pregnancy establishment and that trophoblast-derived VEGF, acting via its specific receptors Flt-1 and KDR, is necessary for blastocyst implantation. The use of cDNA-based expression arrays followed by differential display analysis has provided preliminary understanding of the nature of gene cluster networks operative in the receptive endometrium of potential conception cycles in the rhesus monkey. This knowledge may, in the future, lead to further innovative anti-implantation strategies for targeted pregnancy interception. PMID- 15792649 TI - Religion and medicine. PMID- 15792650 TI - Rationale for decreasing the number of days of the hormone-free interval with use of low-dose oral contraceptive formulations. PMID- 15792651 TI - Unintended pregnancies with the etonogestrel implant (Implanon): a case series from postmarketing experience in Australia. AB - This paper describes a case series of over 200 unintended pregnancies associated with the etonogestrel implant, Implanon. These cases have been reported to the Australian Adverse Drug Reactions Advisory Committee during the first 3 years of marketing in this country. Of 218 cases included, 45 had insufficient data to assess the reason for contraceptive failure and 46 women were determined to have been already pregnant prior to Implanon insertion. Of the remaining 127 cases, the most common reason for unintended pregnancy was failure to insert the implant in 84 women. Other reasons included incorrect timing of insertion (19 cases), expulsion of Implanon (3 cases) and interaction with hepatic enzyme-inducing medicines (8 cases). The remaining 13 cases were classified as product/method failures once other reasons had been excluded. Using the 204,486 Implanon devices subsidized in this period to estimate the population exposed and the 218 pregnancies reported, the approximate failure rate in postmarketing use was 1 in 1000 insertions. These findings (and reports to medical indemnity insurers) have resulted in the development of guidelines and training for doctors inserting Implanon in Australia. PMID- 15792652 TI - Hormonal assessment of women submitted to tubal ligation. AB - Sixteen fertile women aged 30-35 years with regular menstruations were studied before and 6 months after tubal ligation (TL). The diagnosis of ovulation was based on clinical ultrasonographic and hormonal parameters [follicle-stimulating hormone (FSH), luteinizing hormone (LH), E2 and P4]. The menstrual pattern did not vary before and after surgery. Follicular or luteal phase FSH and E2 levels did not differ between the pre- and post-TL period. Luteinizing hormone levels on days -2 and 0 of the follicular phase were significantly higher during the post TL period, while no difference was observed for the luteal phase. P4 levels during the follicular phase did not differ between the two periods, except for day -4, while they were lower during the post-TL luteal phase. The results of the study suggest that in the present patient group, no modifications in the menstrual pattern could be observed 6 months after TL, and TL appears not to interfere with ovulation. Luteinizing hormone levels showed an increase during the ovulatory period after TL, and progesterone secretion decreased during the post-TL luteal phase. PMID- 15792654 TI - Errors in the gross cumulative discontinuation rates for the GyneFix 200 intrauterine system. PMID- 15792655 TI - A reply to D. Wildemeersch's erring reply. Three-year efficacy and acceptability of the GyneFix 200 intrauterine system. PMID- 15792656 TI - In-situ partitioning of butyltin compounds in estuarine sediments. AB - The in-situ solid/pore-water partitioning of tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MBT) was determined for an estuarine sediment profile collected from a commercial marina. Total butyltin levels exceeded sediment quality guideline values, and were 220-8750 microg/kg for TBT, 150-5450 microg/kg for DBT and 130-4250 microg/kg for MBT. Pore-water butyltin concentrations ranged from 0.05 to 2.35 microg/l for TBT, 0.07-3.25 microg/l for DBT, and 0.05-0.53 microg/l for MBT. The partitioning of butyltin compounds between the sediment solid-phase and pore-water was described by an organic carbon normalised distribution ratio (D(OC)). The observed D(OC) values were similar for TBT, DBT and MBT, and were 10(5)-10(6) l/kg. Values for the Butyltin Degradation Index (BDI) were larger than 1 at depths greater than 10 cm below the sediment/water-column interface. This indicates that substantial TBT degradation has occurred in the sediments, and suggests that natural attenuation may be a viable sediment remediation strategy. PMID- 15792657 TI - PM10-bound polycyclic aromatic hydrocarbons (PAHs) in the Greater Area of Athens, Greece. AB - Polycyclic aromatic hydrocarbons (PAHs) associated to ambient PM10 were determined at four sites within the Greater Athens Area (GAA), Greece, during the period May 2001-June 2002. Daily average PM10 samples were collected using reference samplers installed in two urban locations (Maroussi, MAR and Aristotelous, ARI), a mixed urban-industrial location (Elefsina, ELE), and a background location (Thracomacedones, THR). Spatial and temporal variation of ambient PAH levels and possible relationships with conventional air pollutants and meteorological parameters were investigated. Moreover, source identification was attempted using diagnostic ratios. The PAH concentrations observed in this study, even in downtown Athens, were towards the lowest end of the range of values reported for other European urban locations. The mean concentration of the well known carcinogen B[alpha]Py was at all four sites below the proposed value of 1 ng m-3 (annual average assessment threshold). Automobile traffic was identified as the major PAH source in the GAA exhibiting greatest contribution in the urban sites. PMID- 15792658 TI - The hydrochemistry of surface waters in andesitic volcanic area, Norikura volcano, central Japan. AB - Many hydrochemical studies have been reported on surface waters and a major contributor of the water chemistry is considered to be rock dissolution. However, the extent of rock dissolution effect on surface water chemistry has still not been fully understood. The purpose of this research is to clarify the effect to give better understanding on the nature of surface water chemistry. Natural surface water samples were collected from a typical andesitic volcanic area, and major chemicals were investigated in terms of water-rock interaction. The analytical results were subjected to multivariate statistical techniques to understand the geographical distribution and to extract geochemical potential factors affecting the chemical concentration of waters. High correlation between the major chemicals and the altitude of sampling points was found among non geothermal waters. The mole ratios of summation Sigmacations/Si were significantly high in meteoric waters (2.5), low in the surface waters from the summit area of the volcano (0.4), and the median in the surface waters at the mountainside region (1-2) and in geothermal waters (1-2). The basement rocks around the observation areas showed a relatively low ratio (0.4). On the basis of the water chemical compositions, these chemical variation were quantitatively interpreted by the incongruent dissolution reactions for primary minerals in the basement rocks followed by the formation of secondary minerals. The variations of Na+/(Ca2+ + Na+) ratio took positive correlation with [Si] for the non-geothermal waters, and high linear correlation was observed between log(a(Ca2+)/a2(H+)) and log(a(Na+)/a(H+)). These relations were interpreted by an ion-exchange reaction. The factor analysis (FA) gave two major factors to describe the chemical behaviors mentioned above. The contribution proportionate for each factor was calculated as; 65% was the incongruent rock dissolution, and 16% was the ion exchange reaction. PMID- 15792659 TI - Conceptual and numerical model of uranium(VI) reductive immobilization in fractured subsurface sediments. AB - A conceptual model and numerical simulations of bacterial U(VI) reduction in fractured subsurface sediments were developed to assess the potential feasibility of biomineralization at the fracture/matrix interface as a mechanism for immobilization of uranium in structured subsurface media. The model envisions flow of anaerobic groundwater, with or without acetate as an electron donor for stimulation of U(VI) reduction by dissimilatory metal-reducing bacteria (DMRB), within mobile macropores along a one-dimensional flow path. As the groundwater moves along the flow path, U(VI) trapped in the immobile mesopore and micropore domains (the sediment matrix) becomes desorbed and transferred to the mobile macropores (fractures) via a first-order exchange mechanism. By allowing bacterial U(VI) reduction to occur in the mesopore domain (assumed to account for 12% of total sediment pore volume) according to experimentally-determined kinetic parameters and an assumed DMRB abundance of 10(7) cells per cm3 bulk sediment (equivalent to 4 mg of cells per dm3 bulk sediment), the concentration of U(VI) in the macropore domain was reduced ca. 10-fold compared to that predicted in the absence of mesopore DMRB activity after a 6-month simulation period. The results suggest that input of soluble electron donors over a period of years could lead to a major redistribution of uranium in fractured subsurface sediments, converting potentially mobile sorbed U(VI) to an insoluble reduced phase (i.e. uraninite) in the mesopore domain that is expected to be permanently immobile under sustained anaerobic conditions. PMID- 15792660 TI - Comparison of the complexation potential of extracellular polymeric substances (EPS), extracted from activated sludges and produced by pure bacteria strains, for cadmium, lead and nickel. AB - This paper provides information on the metal complexation potential of extracellular polymeric substances (EPS), extracted from activated sludges and from eight pure cultures of bacteria isolated from the same activated sludge. The EPS extracted from pure bacteria cultures are mainly composed of proteins and low quantities of polysaccharides and uronic acids in comparison with EPS extracted from activated sludges. The EPS studied present two apparent pK(a) and the IR spectra show the presence of the same functional groups on all the EPS studied. The ability of EPS to complex Cd, Pb and Ni, was studied at pH 7 with Chau and Ruzic's models using polarography titration. All of the EPS exhibited a greater ability to complex Pb than Ni, Cd showing the weakest affinity overall. The EPS extracted from the pure cultures of bacteria were less able to complex the metals than that extracted from activated sludges. Literature data, IR data and EPS phosphorous content, supported by the EPS pK(a), revealed that carboxylic and phosphoric groups may play a major role in binding to metals at pH 7. This study underlines the importance of metal exposure in order for bacteria to secrete or modify EPS. After exposure, the EPS then exhibit the greatest capacity to bind metal in order to protect bacteria from harmful effects of heavy metals. PMID- 15792661 TI - Sorption of naphthalene derivatives to soils from a long-term field experiment. AB - The influence of long-term farming practices on the soil's behaviour to adsorb hydrophobic organic compounds (HOCs) over long times was investigated. Adsorption of five naphthalene derivatives (naphthalene, 1-naphthol, 1-naphthylamine, 1 hydroxy-2-naphthoic acid, 1,4-naphthoquinone) was examined on soils with varying amounts and origins of soil organic matter obtained after amendment with different organic materials over more than 40 years. Soil organic matter, pore sizes and aggregate stability were significantly altered influencing the adsorption behaviour of the soils. Samples of soil amended with peat having an organic carbon content of 3.4% sorbed naphthalene derivatives stronger than the soil treated with sewage sludge (2.6% C(org)). All other treatments, calcium nitrate, plots without nitrogen fertilizers, grassland, animal manure, green manure and the fallowed soil sorbed less and no significant difference was found between them although the organic carbon content ranged from 1.0% to 2.6%. Thus, a decrease of the carbon content of a soil does not necessarily imply a reduction of sorption capacities for hydrophobic compounds such as naphthalene derivatives. Furthermore, the importance of protonation of HOCs for the adsorption on soil surfaces was shown. Different polarities of electronic structures of HOCs distinctly influence their adsorption behaviour. PMID- 15792662 TI - Use of 15N-depleted artificial compost in bound residue studies. AB - Association of bound residues to soil humic matter may be accomplished by different binding mechanisms such as sequestration in hydrophobic interiors of the organic material or covalent linkage to the organic matter. The structures and chemical environments of compounds can be observed by NMR spectroscopy. We applied 15N-NMR spectroscopy to study the soil-bound residues of 15N-labeled simazine. As the 15N-isotope has a low sensitivity and natural abundance 15N-NMR experiments require long measurement times and often result in low signal-to noise (S/N) ratios. Therefore, in addition to the use of 15N-labeled simazine, 15N-depleted compost was used to reduce the amount of background signal and enhance the sensitivity. The compost was produced from maize and wheat plants grown on sand with 15N-depleted NH4NO3 as sole nitrogen source. The plants were freeze-dried, ground and mixed with sand for composting. After a composting period of 224 days analysis of the compost revealed a 15N-content of 267 ppm as opposed to a natural abundance of 3650 ppm. Characterization of this artificial compost produced parameter values similar to those of a natural compost. The 13C NMR-spectra of the humic and fulvic acids during different stages of maturity showed that there was a shift from single-bond functional groups to more complex double-bond and aromatic structures. Experiments with this compost showed an increased signal intensity. The improved sensitivity made it possible to obtain interpretable NMR signals in contrast to experiments with 15N-simazine on native soil where no signals were detectable. The data indicated that the bound residues of simazine are composed of metabolites resulting from N-dealkylation and triazine ring destruction. Silylation of the bound residues showed a very strong binding of the residues to the matrix as only a small fraction could be solubilized. PMID- 15792663 TI - Effect of pH on the aggregation of a gray humic acid in colloidal and solid states. AB - Gray humic acids have a marked colloidal character, a large number of surface functional groups, and are subject to aggregation phenomena. They are able to complex soluble pollutants, and initiate flocculation processes as a function of environmental conditions. The aim of this work is to study the aggregation of a gray humic acid, which is stable in colloidal dispersion, by means of photon correlation spectroscopy, and molecular modeling. The effect of this aggregation in the solid state is also studied by means of N2 (to 77 K) and CO2 (to 273 K) adsorption isotherms, as well as FT-IR absorption. The variation of the colloid's zeta potential and size, with pH, reflects the ionization of the carboxylic and phenolic acidic groups, and a linear dependence of size on zeta potential. The decrease in the size of the colloids seems to be more affected by the ionization of the phenolic acid groups, than by that of the carboxylic acid groups, which is likely because in the case of the ionized carboxylic groups the humic colloids are still capable of generating H-bonds. In the solid state, aggregation effects are illustrated by a decrease in surface area, and a disappearance of certain micropores, with increasing pH. These features are likely due to an inhibition of aggregation in the colloidal state as a consequence of the increase in charge that results from ionization of the acidic groups, and also to an increased hindrance to H-bond formation, due to the loss of protons during the above mentioned ionization process. PMID- 15792664 TI - Levels of total suspended particulate matter and major trace elements in Kosovo: a source identification and apportionment study. AB - Concentration levels of total suspended particles (TSP) and 27 major, minor and trace elemental components were determined at four sites in Kosovo through a 1 year survey (January-December 2002). Ambient concentrations were evaluated in comparison to limit values. The origin of elemental TSP constituents was investigated by calculating enrichment factors and diagnostic ratios. Multivariate statistics, such as hierarchical cluster analysis and factor analysis, were also employed to identify emission sources. A multivariate statistical receptor model (Absolute Principal Component Analysis, APCA) was applied to quantify source contributions. Soil dust, cement production, vehicular emissions, brake wear, and fuel combustion were identified as major sources with variable contributions at the four sampling sites. PMID- 15792665 TI - Measurement of phase-distributed nitrophenols in Rome ambient air. AB - Atmospheric nitrophenols (NPs) were determined both in the gas and particle phases by combining the annular denuder sampling technique with GC-MS analysis. The phase distribution of six mono-NPs identified in air sampled in downtown Rome was assessed. 4-NP, 3-methyl-4-NP and 2,6-dimethyl-4-NP were found for more than 75% in the particle phase, whilst 2-NP, 4-methyl-2-NP and 5-methyl-2-NP predominated in the gas phase. Concentration levels lower than 20 ng m-3 were observed for all NPs, among which 4-NP (17.8+/-5.6 ng m-3) and 2-NP (10.4+/-4.2 ng m-3) were the most abundant congeners in the two phases, respectively. PMID- 15792666 TI - Photodegradation of phosphonates in water. AB - Phosphonates are widely used as chelating agents, e.g., in water cooling systems, in bleaching baths or as scale inhibitors in deflocculation agents. They are considered to be difficult to degrade and produce aminomethylphosphonic acid (AMPA) as a metabolite. As the fate of phosphonates in the environment is not very well known the present work aims at simulating the time dependent photodegradation of four selected phosphonates: nitrilotris-methylenephosphonic acid (NTMP), ethylenediamine-tetra-methylenephosphonic acid (EDTMP), diethylenetriaminepenta-methylenephosphonic acid (DTPMP) and hexaethylenediamine tetra-methylenephosphonic acid (HDTMP), at concentrations of 1 mg/l (i.e. 3.2 microM NTMP, 2.3 microM EDTMP, 1.7 microM DTPMP and 2.0 microM HDTMP) irradiated by a middle pressure mercury lamp emitting between 190 and 600 nm. The influence of iron under different pH ranges (3, 5-6 and 10) are tested. The degradation of phosphonates is measured by the release of orthophosphates (PO4-P) and aminomethylphosphonic acid (AMPA). This study shows that phosphonates are substances that undergo UV light conversion, which is enhanced in the presence of iron. The half-life without iron is between 15 and 35 min at pH 3, between 10 and 35 min at pH 5-6 and between 50 and 75 min at pH 10. The half-life in the presence of 3.6 microM iron is between 5 and 10 min at pH 3, between 5 and 15 min at pH 5-6 and between 35 and 60 min at pH 10. The individual substances do not significantly influence the reaction rates whereas the presence of iron and the pH have significant effects. The total conversion of phosphonates after 90 min is 75-100% for pH values of 3 and 5-6 and 55-75% for a pH of 10 dependent on the presence of iron. In the environment longer degradation times are to be expected since natural light is weaker by a factor between 125 and 300 in the UVB, a factor between 3 and 8 in the UVA and of the same intensity in the visible range than the light in our study. Although orthophosphates are the major products, AMPA is also shown to be a by-product of the photodegradation of phosphonates that is later converted into orthophosphate. PMID- 15792667 TI - Potential assessment of the "support vector machine" method in forecasting ambient air pollutant trends. AB - Monitoring and forecasting of air quality parameters are popular and important topics of atmospheric and environmental research today due to the health impact caused by exposing to air pollutants existing in urban air. The accurate models for air pollutant prediction are needed because such models would allow forecasting and diagnosing potential compliance or non-compliance in both short- and long-term aspects. Artificial neural networks (ANN) are regarded as reliable and cost-effective method to achieve such tasks and have produced some promising results to date. Although ANN has addressed more attentions to environmental researchers, its inherent drawbacks, e.g., local minima, over-fitting training, poor generalization performance, determination of the appropriate network architecture, etc., impede the practical application of ANN. Support vector machine (SVM), a novel type of learning machine based on statistical learning theory, can be used for regression and time series prediction and have been reported to perform well by some promising results. The work presented in this paper aims to examine the feasibility of applying SVM to predict air pollutant levels in advancing time series based on the monitored air pollutant database in Hong Kong downtown area. At the same time, the functional characteristics of SVM are investigated in the study. The experimental comparisons between the SVM model and the classical radial basis function (RBF) network demonstrate that the SVM is superior to the conventional RBF network in predicting air quality parameters with different time series and of better generalization performance than the RBF model. PMID- 15792668 TI - An evaluation of electrode insertion techniques for measurement of redox potential in estuarine sediments. AB - E(h) measurements by electrodes are commonly used to characterize redox status of sediments in freshwater, marine and estuarine studies, due to the relative ease and rapidity of data collection. In our studies of fine-grained estuarine seabeds, we observed that E(h) values measured in intact sediment cores were influenced by different electrode insertion techniques. Sediment E(h) measurements generated via lateral insertion of platinum electrodes through silicone-filled ports in acrylic cores were systematically more positive (on the order of 10-100 mV) than profiles generated via vertical insertion of platinum electrodes downward through the sediment-water interface of the same cores. A review of the literature indicated that while researchers routinely use both insertion techniques to measure E(h), no discrepancy in output has previously been reported. We discuss the results of three experiments conducted to determine if the discrepancy in output was caused by electrode poisoning by sulfides during the stepwise vertical insertion technique, or was caused by contact of the electrode with the silicone plug during the lateral insertion technique. We conclude that contact between the platinum surface of the electrode and the silicone plug biases the E(h) measurements, resulting in erroneously positive E(h) values. Insertion of electrodes into sediment through silicone plugs produced E(h) values that were an average of 105.6 mV (+/-10.4 SE) more positive than values generated upon electrode insertion directly into sediment. Thus, we recommend against using an insertion technique where the platinum electrode remains in contact with the silicone plug, as this method results in misclassification of sediment redox state and estimated depth of the redoxcline. PMID- 15792669 TI - An alternative method for predicting organic solute volatilization rates under gas and liquid turbulence. AB - A method for predicting organic compound volatilization rates under turbulent liquid and gas conditions is developed. The reference compounds are classified according to their physico-chemical properties. The mass transfer coefficient (K(OL)) ratios for organic solutes to the reference compounds are constant for a wide range of environmental conditions, including liquid or air turbulence, or both at once. The obtained results indicate that when the environmental conditions are the same the determination of the volatilization rates is strongly dependent on the solute properties and the chemical structure. The presented method can more effectively estimate the volatilization rates of the solutes than the traditional one under various environmental conditions especially for low volatility solutes. The advantages and disadvantages of the traditional method are also discussed. PMID- 15792670 TI - Effects of dissolved organic carbon on sorption of 3,4-dichloroaniline and 4 bromoaniline in a calcareous soil. AB - To evaluate the effects of dissolved organic carbon on sorption of 3,4 dichloroaniline (3,4-DCA) and 4-bromoaniline (4-BA) on soils, batch sorption experiments were carried out. The soil used was a typical calcareous soil from south-eastern Spain. Two different types of dissolved organic carbon were used, that is, dissolved organic carbon extracts from a commercial peat (DOC-PE) and high-purity tannic acid (DOC-TA). The experiments were carried out in a 0.01 M CaCl2 aqueous medium at 25 degrees C. The results obtained from the sorption experiments show that the presence of both DOC-PE and DOC-TA, over a concentration range of 15-100 mg l-1, produced in all cases, an increase in the amount of 3,4-DCA and 4-BA adsorbed on the soil studied. PMID- 15792671 TI - Evaluating factors that influence microbial phenanthrene biodegradation rates by regression with categorical variables. AB - To advance the accuracy of bioremediation measurements, it is useful before specific experiments to attribute or estimate the influence of both experimental as well as field conditions on the expected magnitudes of microbial degradation rate coefficients. This paper analyzes the numerical contribution, or influence, of categories of conditions, such as bacterial adaptive state, electron acceptor type, mixing, generalized sorption conditions, and biodegradation temperature, on published phenanthrene biodegradation rates as an example of our regression approach. A fundamental microbial degradation rate equation is transformed to an additive model, then using multiple linear regression on published data, coefficients (of categorical variables) and a linear model are presented that estimate first-order biodegradation rate coefficients to within a factor of 3. Numerical estimates of how much bacterial adaptive state and presence of a sorption phase, the two most statistically significant factors, alter the phenanthrene biodegradation rate are presented. The influence of some measurement or field conditions, for example, the influence of oxygen reduction versus optimal nitrate reduction, cannot be distinguished statistically given the available data and range. The regression model is tested using conditions from newly published papers to estimate a priori the expected rate, which compares very favorably to measurements reported in the papers. Due to limited published data and range for extreme cases, the current coefficients do not apply to degradation of very aged phenanthrene nor very low concentrations of electron acceptors. As estimating tools, however, the coefficients themselves and the regression approach have very beneficial roles in design of experiments for both laboratory and field settings. Our method can be applied to other PAHs as sufficient data become available. PMID- 15792672 TI - Modeling interactions at the tributyltin-kaolinite interface. AB - Tributyltin (TBT) is a common pollutant in natural environments and the interaction with mineral surfaces largely determine its solubility, speciation, bioavailability, and transport in aqueous systems. The present work aimed at quantifying the TBT adsorption using kaolinite and a kaolinite-rich sediment as sorbent materials. Experiments were conducted under conditions that are important from an environmental perspective. Proton adsorption data were determined for kaolinite as a function of pH and electrolyte concentrations to ascertain intrinsic acidity constants and site density values. The pH(zpc) of kaolinite sample KGa was determined at 4.9 by surface titration. The generalized diffused double layer model (DLM) was used to quantify both, proton and TBT adsorption. Following intrinsic acidity and TBT binding constants resulting from the TBT/kaolinite system were used: >SOH ==> >SO- + H+, logK = -5.4; >SOH + H+ ==> SOH2+, logK = 4.6; >XNa + H ==> XH + Na+, logK(X/H+) = -1.1; >SO- + TBT+ ==> >SOTBT, logK = 3.5; >XNa + TBT+ ==> Na+, logK(X/TBT) = 1.0. All surface-active variable charge sites on kaolinite, namely >AlOH and >SiOH are grouped and collectively refer to as >SOH in this paper. >XNa refers to ion exchange sites. Modeling of TBT adsorption onto kaolinite was conducted distinguishing selective (high affinity) sites (>S(S)OH) in addition to non-selective sites (>SOH). The inclusion of >S(S)OH was essential in order to quantify TBT adsorption successfully, while the inclusion of >XNa was optional. The reduction of surface coverage values by a 10-fold TBT adsorption modeling is in agreement with the results of molecular model calculations of the system. Parameters calculated for the monophase kaolinite were subsequently used to quantify the TBT adsorption onto kaolinite-rich sediment. PMID- 15792673 TI - Historical perspective of factor XI. PMID- 15792674 TI - Optimisation of the prothrombin complex concentrate dose for warfarin reversal. PMID- 15792675 TI - Optimal dose of prothrombin complex concentrate for acute reversal of oral anticoagulation. AB - We investigated optimal dose of prothrombin complex concentrate (PCC) for acute reversal of oral anticoagulation in patients with major hemorrhagic complications or who required invasive procedures. We also checked how rapidly international normalized ratio (INR) was reversed after PCC administration. INR was measured before and 10-60 min after administration of PCC with or without vitamin K in 42 patients (men 28, women 14, median age of 70 years old) who had received warfarin but required rapid reversal of INR because of a hemorrhagic complication or medical procedure. The amount of PCC administered was 200 IU in six patients, 500 IU in 30, 1000 IU in 3, and 1500 IU in the other 3. Additional administration of PCC was performed when the correction of INR was inadequate. In 10 of the 42 cases, INR was measured serially, before, 10 and 60 min and 12-24 h after the administration of PCC and vitamin K. In the six patients who received PCC of 200 IU, INR values of 3.34 median (range 2.06 to 5.08) decreased to 1.85 (range 1.23 to 2.43) significantly (Wilcoxon's rank sum test, p=0.028), but in three patients (50%), INR values were still above 2.0 after the administration. In 30 patients treated with PCC of 500 IU, values decreased from 2.49 median (range 1.54 to 10.00) to 1.19 (range 0.87 to 1.55) significantly (p<0.0001). The corrected INR values were below 1.5 in 25 of 26 patients (96%) who had initial INR values from 2.0 to 4.9. In four patients with initial INR of 5.0 or more, the reversed INR was below 1.5 in one (25%), between 1.5 and 2.0 in two (50%), and above 2.0 in one (25%) who had additional administration of 500 IU PCC lowering INR from 2.01 to 1.48. Values of INR in the six patients receiving 1000 IU or 1500 IU, INR decreased from 2.33 median (range 1.96 to 4.00) to 0.96 (range 0.87 to 1.24, p=0.028). In the 10 patients with serial measurement, INR changed from 2.67 median (range 2.05 to 10.00) to 1.17 (range 0.99 to 1.60) 10 min after the administration. The INR values remained stable 60 min and 12-24 h after the PCC administration. The 500 IU of PCC is likely to be optimal dose of PCC for emergent reversal of INR in patients requiring rapid correction of INR below 5.0, but to be inadequate dose in patients with INR of 5.0 or more. PCC administration with vitamin K may finish reversing INR rapidly within 10 min and keep the reversed INR values for 12-24 h. PMID- 15792676 TI - Coagulation status and complications of pregnancy. AB - INTRODUCTION: There is much interest in the relationship between coagulation status and complications of pregnancy. The thrombelastograph (TEG) has been proposed as a useful, inexpensive tool to screen for patients with hypercoagulable states. MATERIALS AND METHODS: We investigated 588 unselected pregnant women at booking, obtaining blood samples for TEG and thrombophilia investigation. Pregnancy outcome data was recorded. RESULTS: We found significant correlations between TEG parameters and the Prothrombin time (PT) and Activated Partial Thromboplastin time (APTT) (p<0.01) and with plasma Antithrombin level (p<0.01). There was no correlation between TEG and other thrombophilic defects (protein C, protein S, Factor V Leiden mutation, Prothrombin G20210A mutation, MTHFR C677T mutation and Lupus Anticoagulant). There was a significant association of TEG parameters with mid-trimester loss (MTL) but not with other adverse pregnancy outcomes. CONCLUSIONS: The correlation between TEG and PT, APTT and antithrombin level supports its value in providing a global measure of haemostasis. Coagulation status at booking is associated with increased risk of MTL but not with complications occurring later in pregnancy. PMID- 15792677 TI - Atorvastatin reduces thrombin generation after percutaneous coronary intervention independent of soluble tissue factor. AB - INTRODUCTION: Statins were previously shown to suppress cellular tissue factor (TF) in vitro. Here, we investigated the effect of atorvastatin on the TF-pathway and thrombin generation after coronary angioplasty and stenting in vivo. MATERIALS AND METHODS: A cohort of 30 patients with coronary artery disease (CAD) was randomised to treatment with either none (n=10), 10 mg (n=10) or 80 mg (n=10) atorvastatin per day for the postinterventional period of 6 months starting the day before percutaneous coronary intervention (PCI). Fasting blood samples were collected on admission and after 6 weeks and 6 months of statin therapy to determine sTF, free tissue factor pathway inhibitor (TFPI) and prothrombin fragment F1.2 by immunoassay. RESULTS: Soluble TF (sTF) significantly correlated with thrombin generation as measured by prothrombin fragment F1.2 at baseline. This correlation was lost 6 weeks and 6 months after initiation of statin therapy. In vivo, F1.2 was significantly lowered after 6 months of statin therapy by both, low dose (0 vs. 10 mg: 1.3+/-0.3 vs. 0.7+/-0.2 ng/ml; P<0.05) and high dose (0 vs. 80 mg: 1.2+/-0.3 vs. 0.6+/-0.2 ng/ml; P=0.01) atorvastatin compared to control. However, sTF and free TFPI did not change significantly with atorvastatin therapy when compared to baseline or control. CONCLUSIONS: Our results demonstrate reduced in vivo generation of thrombin six months after percutaneous coronary intervention and statin therapy independent of sTF and free TFPI. PMID- 15792678 TI - Incidence of heparin-PF4 complex antibody formation and heparin-induced thrombocytopenia in acute coronary syndrome. AB - A multicenter prospective study on the rate of seroconversion of antibodies to heparin-PF4 complexes (heparin-induced thrombocytopenia [HIT] antibodies) during and after heparin treatment for 4 weeks was carried out in Japanese patients with acute coronary syndrome (ACS). A total of 254 ACS patients treated with heparin were enrolled consecutively from 12 facilities of cardiology. Two patients with preexisting HIT antibodies were excluded from the analysis. The total seroconversion rate for four weeks during and after heparin treatment was 8.7% (n=22, 95% confidence interval [CI]: 5.9-13.1), including values of 3.2% (n=8) at the end of heparin infusion and 5.5% (n=14) at 4 weeks. Among 22 seroconverted patients, four developed HIT and two of the four had the complication of thrombosis. The incidence of HIT was 1.6% (n=4, 95% CI: 0.04-3.1). The risk for thromboembolic development was higher in the seroconverted patients (odds ratio, 17.4, 95% CI: 5.2-58.4, p<0.0001) than nonconverted patients. An analysis of factors affecting the seroconversion rate was carried out. The seroconversion rate for ACS patients who underwent percutaneous coronary intervention (PCI; n=163) was 12.3%, significantly higher than the 2.3% in patients who did not undergo PCI (n=89), leading to an odds ratio of 6.1 (95% CI: 1.4-26.7, p=0.009). A significant odds ratio was obtained for each factor affecting the seroconversion: 3.5 (95% CI: 1.3-9.9, p=0.014) for more than 5 days of heparin infusion, 3.0 (95% CI: 1.2-7.6, p=0.035) for a thrombotic history and 2.7 (95% CI: 1.1-6.8, p=0.039) for hyperlipidemia. No other factor, including age or diabetes mellitus, contributed to the seroconversion. Therefore, PCI, duration of heparin treatment and thrombotic history facilitated the seroconversion in ACS patients. PCI patients treated for more than 5 days with heparin showed a maximal seroconversion rate of 18.3% (95% CI: 13.8-22.2). This high rate in PCI patients did not interact with age, type of underlying disease of unstable angina or myocardial infarction or thrombotic history. In conclusion, ACS patients demonstrating seroconversion are at risk of thromboembolic development due to the likelihood of immunomediated endothelial dysfunction. The increase in the rate of seroconversion in ACS patients would be affected by factors such as PCI with mechanical stress, longer duration of heparin treatment, thrombotic history and presence of hyperlipidemia. If PCI is undertaken with heparin anticoagulation for more than 5 days, seroconversion would easily occur, and the seroconverted patients could subsequently suffer from HIT. PMID- 15792679 TI - Levels of the adipocyte-derived plasma protein, adiponectin, have a close relationship with atheroma. AB - INTRODUCTION: Inflammation is a key process in atherosclerotic formation. Structural changes in the carotid arterial wall including detection of focal plaques are measured as the intima-media thickness (IMT) providing an index of atheroma. Coronary arterial plaques may be considered as vascular structural changes. Distensibility of the arteries can be assessed by functional changes in pulse wave velocity (PWV) providing an index of sclerosis. Adiponectin has potential antiatherosclerotic properties. We hypothesized that adiponectin was associated with atherosclerotic vascular changes involved in inflammation. MATERIALS AND METHODS: We enrolled 142 patients with coronary artery disease (CAD) and 108 control patients, matched for age, sex, and body mass index (BMI) with CAD patients. We investigated the relationship between adiponectin, C reactive protein (CRP), and atherosclerotic vascular changes. RESULTS: CRP (p=0.0009), high-density lipoprotein cholesterol (HDL-C; p=0.02), and IMTmax (p=0.02) were determinants of adiponectin independent of glucose intolerance (p=0.0001), BMI (p=0.002), and CAD (p=0.03), all of which have been significantly associated with adiponectin (r=0.38). Adiponectin was not correlated with PWV. CRP, glucose intolerance, and HDL-C that correlated with adiponectin were inversely correlated with IMTmax and CAD. CRP was negatively correlated with HDL C (r=-0.24, p=0.0002) and positively correlated with glucose intolerance (r=0.15, p=0.01). CONCLUSIONS: Adiponectin has a close relationship with CRP, IMTmax, CAD, HDL-C, and other established risk factors. CRP, glucose intolerance, and HDL-C are common mediators between adiponectin and atheromatous vascular changes, which are contrary to each other. The exacerbation of atherogenesis may be involved in a decrease of adiponectin through abnormal glyco- and lipid-metabolism by promoting inflammation. PMID- 15792680 TI - Activated protein C resistance and low molecular weight lipoprotein (a): dual pathogens for atherothrombosis? PMID- 15792681 TI - Inhibitory effects of sulphated flavonoids isolated from Flaveria bidentis on platelet aggregation. AB - Flaveria bidentis is a plant species that has as major constituents sulphated flavonoids in the highest degree of sulphatation. Among them, quercetin 3,7,3',4' tetrasulphate (QTS) and quercetin 3-acetyl-7,3',4'-trisulphate (ATS) are the most important constituents. Both showed anticoagulant properties. The objective of the present study was to evaluate the effects of these flavonoids on human platelet aggregation in comparison with the well-known inhibitor quercetin (Qc) by using several agonists. Platelet-rich plasma (PRP) or washed human platelets (WP) were incubated with different concentrations of the flavonoids to be tested (1 to 1000 microM, final concentration), and the platelet aggregation was induced by using adenosine 5'-diphosphate (ADP), epinephrine (EP), collagen, arachidonic acid (AA) and ristocetin as agonists. QTS (500 microM) and Qc (250 microM) markedly inhibited platelet aggregation with all the aggregant agents, except ristocetin, whereas ATS (1000 microM) showed only slight antiplatelet effects. In addition, QTS and Qc antagonized the aggregation of PRP or WP induced by U-46619, a mimetic thromboxane A2 (TxA2) receptor agonist. Challenged with collagen or arachidonic acid, the thromboxane B2 (TxB2) formation was also inhibited by the flavonoids, mainly by QTS and Qc, in WP. These results demonstrate that QTS and in minor extension ATS induce a deleterious effect on the production of TxA2, as judged by TxB2 formation, in stimulated WP and a marked interference on the TxA2 receptor according to the profile of inhibition of the agonist-induced platelet aggregation when using ADP, EP, AA and collagen and confirmed with U-46619. PMID- 15792682 TI - A defined peptide that inhibits the formation of the glycoprotein IIb and IIIa complex. AB - Collagen-platelet interaction plays an important role in hemostasis and pathological thrombosis. The proposed mechanism of the interaction was the activation of platelets-->releasing of contents from granules-->aggregation. The common end point is the platelets and fibrin aggregates. Platelet glycoprotein (GP) IIb/IIIa (the alphaIIbbeta3 integrin) complexes serve as a receptor for the binding of fibrinogen to form firmed aggregates. Blockading of GP IIb/IIIa has been proposed to prevent platelet aggregation independent of the substance(s) responsible for activating the platelets. The development of various forms of GP IIb/IIIa inhibitor has resulted in the inhibition of platelet aggregation, although studies of alphaIIbbeta3 receptor function and various GP IIb/IIIa inhibitors have demonstrated the potential for these agents to produce effects on other aspects of platelet function as well as having nonplatelet effects. This study investigated platelet inhibition provided by blocking the GP IIb/IIIa complex formation by using a peptide derived from the GP IIIa molecule. The peptide inhibits both types I and III collagen-induced platelet aggregation in a dose-dependent manner. The defined peptide interferes with the formation of the GP IIb/IIIa complex by inhibiting the binding of FITC-PAC-1 onto ADP-, type I collagen-, and type III collagen-activated platelets. However, P-selectin secretion is not affected by the peptide. In addition, the peptide is not interfering with the binding of FITC-PAC-1 to platelets that were preincubated with indomethacin. Results from this study may suggest that the defined peptide is an effective agent to block the interaction of types I and III collagen with platelets. PMID- 15792683 TI - Effect of subcutaneous administration of dalteparin on lupus anticoagulant assays. AB - INTRODUCTION: Treatment with unfractionated heparin (UH) is known to affect screening tests for lupus anticoagulant (LA). False positive test results are common because confirmatory steps lack sufficient specificity to distinguish between LA and the presence of heparin. In this study, we wanted to see if therapeutic levels of low-molecular weight heparin (LMWH) may cause false positive tests for LA or alter the LA test results in LA-positive patients. We also wanted to evaluate the need to include heparin-neutralizing agents in the reagents. MATERIALS AND METHODS: Six healthy subjects without LA and six LA positive patients were given 100 IU/kg dalteparin subcutaneously (s.c.). Samples for three in-house and two commercially available LA tests were taken before and 4 h after the injection. LA test results were calculated as normalized screening/confirm ratios or as recommended by the manufacturers. RESULTS: With both healthy subjects and LA patients, only small and clinically unimportant differences in mean clotting times and final test results were seen 4 h after subcutaneous dalteparin injections, at anti-FXa activities within the therapeutic range. CONCLUSIONS: Our study with dalteparin suggests that LMWH therapy with plasma concentrations within the therapeutic range does not cause false positive tests for LA when normalized screening/confirm ratios are applied; nor do test results for LA-positive patients seem to be significantly altered. Heparin neutralizing agents did not influence test performance. PMID- 15792684 TI - Coagulation Inhibitor Potential: a study of assay variables. AB - INTRODUCTION: The Coagulation Inhibitor Potential (CIP) assay is a further development of the Overall Haemostatic Potential (OHP) by He et al. which monitors fibrin aggregation in plasma activated by thrombin, tissue-type plasminogen activator (t-PA) and CaCl2. CIP reflects the ability of enhanced coagulation inhibitors, antithrombin (AT) and protein C to counteract slow in vitro coagulation. MATERIALS AND METHODS: Plasma from 23 persons with different types of severe thrombophilic conditions were compared to normal controls. Pentasaccharide and Protac were added to enhance inhibition of coagulation. The relative effect of enhanced inhibition was calculated. RESULTS: The assay performed equally well with tissue factor (TF) and thrombin as triggers of coagulation (P<0.0001). Protac and rabbit thrombomodulin (TM) both demonstrated an anticoagulant effect, but only when pentasaccharide was present. Rabbit TM reduced fibrinolysis and Protac was used in the standard procedure. Pentasaccharide and Protac had a synergistic effect resulting in a clearly reduced coagulation in the normal controls, but less in persons with thrombophilia. Receiver operator characteristic (ROC) analysis indicated that at 100% sensitivity, specificity was 78% with and without t-PA. CONCLUSIONS: The improvement of the CIP by omitting t-PA allows a more rapid testing. There is a need for evaluating the CIP assay in larger patient groups. PMID- 15792686 TI - First report of a FVII-deficient Indian patient carrying double heterozygous mutations in the FVII gene. PMID- 15792685 TI - Evaluation of two experimental venous thrombosis models in the rat. PMID- 15792687 TI - Toll-like receptors on platelets: the key for disseminated intravascular coagulation in sepsis? PMID- 15792688 TI - Parasitic manipulation: where are we and where should we go? PMID- 15792689 TI - Parasitic manipulation: should we go anywhere? PMID- 15792690 TI - Parasitic manipulation: further considerations. PMID- 15792691 TI - Manipulations: variations on the themes of signalling and exaptation. PMID- 15792692 TI - Trophic transmission of parasites and host behavior modification. PMID- 15792693 TI - Parasite manipulation of host behavior: mechanisms, ecology, and future directions. PMID- 15792694 TI - Studying adaptive changes in the behaviour of infected hosts: a long and winding road. PMID- 15792695 TI - Parasitic manipulation: going beyond behaviour. PMID- 15792696 TI - Parasite manipulation: stretching the concepts. PMID- 15792697 TI - Do all parasites manipulate their hosts? PMID- 15792698 TI - The true cost of host manipulation by parasites. PMID- 15792699 TI - Parasite-host conflicts: winners and losers or negotiated settlements? PMID- 15792700 TI - Parasitic manipulation: a theoretical framework may help. PMID- 15792701 TI - The proteomics: a new prospect for studying parasitic manipulation. PMID- 15792702 TI - Parasitic manipulation of host life history and sexual behaviour. PMID- 15792703 TI - Conflict between co-occurring parasites as a confounding factor in manipulation studies? PMID- 15792704 TI - Parasitic manipulation: a social context. PMID- 15792705 TI - The extended manipulation: role of parasitism in cleaning symbiosis. PMID- 15792706 TI - Malaria as a manipulator. PMID- 15792707 TI - Parasitic manipulation: where else should we go? PMID- 15792708 TI - Look what the cat dragged in: do parasites contribute to human cultural diversity? PMID- 15792711 TI - An in situ silver cationization method for hydrocarbon mass spectrometry. AB - We have developed a novel cationization method for the analysis of long-chain hydrocarbons via UV laser desorption mass spectrometry. In this technique we electrospray a thin coating of AgNO3 over a sample and perform UV laser desorption to produce Ag+ cationization of sample molecules. Use of this technique in our microscope/TOF-MS allows us to determine the spatial distribution of the species we detect in the sample. We demonstrate 8-mu spatial resolution, and submicron resolution is possible in principle. In mixed samples containing aromatic and aliphatic compounds, the aromatic compounds ionize directly and do not form adducts, and thus give single peaks as opposed to doublets from silver cations. This enables distinction between aromatic and aliphatic compounds that are in the same sample. PMID- 15792709 TI - Manipulation: expansion of the paradigm. PMID- 15792712 TI - A minimum thickness gate valve with integrated ion optics for mass spectrometry. AB - A minimum thickness gate valve design for mass spectrometry is described in detail. The ion optics required to transmit ions from the source to the ICR cell are integrated into the design to minimize fringe field effects on the ions as they travel through the gate valve. The total thickness of the complete gate valve assembly is 1.03 in. (26.2 mm) with a maximum fringe field distance of 0.065 in. (1.7 mm). The gate valve is able to maintain a vacuum of <10(-10) mbar at the ICR cell when the source is vented to atmosphere and the estimated ion transfer efficiency is >95%. PMID- 15792713 TI - The relative influences of acidity and polarity on responsiveness of small organic molecules to analysis with negative ion electrospray ionization mass spectrometry (ESI-MS). AB - The purpose of the work presented here was to evaluate the influence of solution composition and analyte characteristics on responsiveness to analysis with negative ion electrospray ionization mass spectrometry. The responses of a series of structurally diverse acidic molecules were compared in various solvents. Response was generally observed to be higher in methanol than acetonitrile and response for all analytes was poorer when water was mixed with the organic solvent. A positive correlation between negative ion ESI-MS response and log P was observed when either acetonitrile or methanol was used as the electrospray solvent. This result was expected because analytes with significant nonpolar character should be particularly responsive to ESI-MS analysis due to their higher affinity for electrospray droplet surfaces. It was also predicted that highly acidic analytes would be most responsive to analysis with negative ion ESI MS due to their tendency to form negative ions. However, for the analytes studied here, acidity was found not to have a consistent influence on ESI-MS response. Many of the highly acidic molecules were quite polar and, consequently, were poorly responsive. Furthermore, the deprotonated molecular ion was detected for a number of molecules with very high pKa values, which would not be expected to form negative ions in the bulk solution. Ultimately, these results indicate that acidity is not a conclusive parameter for prediction of the relative magnitudes of negative ion ESI-MS response among a diverse series of analytes. Analyte polarity does; however, appear to be useful for this purpose. PMID- 15792714 TI - Bacterial analysis by MALDI-TOF mass spectrometry: an inter-laboratory comparison. AB - Bacterial analysis by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry has been demonstrated in numerous laboratories, and a few attempts have been made to compare results from different laboratories on the same organism. It has been difficult to understand the causes behind the observed differences between laboratories when different instruments, matrices, solvents, etc. are used. In order to establish this technique as a useful tool for bacterial identification, additional efforts in standardizing the methods by which MALDI mass spectra are obtained and comparisons of spectra from different instruments with different operators are needed. Presented here is an extension of our previous single-laboratory reproducibility study with three different laboratories in a controlled experiment with aliquots of the same bacterial culture, matrix stock solution, and calibrant standards. Using automated spectral collection of whole-cell bacteria and automated data processing and analysis algorithms, fingerprints from three different laboratories were constructed and compared. Nine of the ions appeared reproducibly within all three laboratories, with additional unique ions observed within each of the laboratories. An initial evaluation of the ability to use a fingerprint generated within one laboratory for bacterial identification of a sample from another laboratory is presented, and strategies for improving identification rates between laboratories is discussed. PMID- 15792715 TI - Improved mass accuracy for tandem mass spectrometry. AB - With the emergence of top-down proteomics, the ability to achieve high mass measurement accuracy on tandem MS/MS data will be beneficial for protein identification and characterization. (FT-ICR) Fourier transform ion cyclotron resonance mass spectrometers are the ideal instruments to perform these experiments with their ability to provide high resolution and mass accuracy. A major limitation to mass measurement accuracy in FT-ICR instruments arises from the occurrence of space charge effects. These space charge effects shift the cyclotron frequency of the ions, which compromises the mass measurement accuracy. While several methods have been developed that correct these space charge effects, they have limitations when applied to MS/MS experiments. It has already been shown that additional information inherent in electrospray spectra can be used for improved mass measurement accuracy with the use of a computer algorithm called DeCAL (deconvolution of Coulombic affected linearity). This paper highlights a new application of the strategy for improved mass accuracy in tandem mass analysis. The results show a significant improvement in mass measurement accuracy on complex electron capture dissociation spectra of proteins. We also demonstrate how the improvement in mass accuracy can increase the confidence in protein identification from the fragment masses of proteins acquired in MS/MS experiments. PMID- 15792716 TI - Microwave-assisted acid hydrolysis of proteins combined with liquid chromatography MALDI MS/MS for protein identification. AB - Simple and efficient digestion of proteins, particularly hydrophobic membrane proteins, is of significance for comprehensive proteome analysis using the bottom up approach. We report a microwave-assisted acid hydrolysis (MAAH) method for rapid protein degradation for peptide mass mapping and tandem mass spectrometric analysis of peptides for protein identification. It uses 25% trifluoroacetic acid (TFA) aqueous solution to dissolve or suspend proteins, followed by microwave irradiation for 10 min. This detergent-free method generates peptide mixtures that can be directly analyzed by liquid chromatography (LC) matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) without the need of extensive sample cleanup. LC-MALDI MS/MS analysis of the hydrolysate from 5 microg of a model transmembrane protein, bacteriorhodopsin, resulted in almost complete sequence coverage by the peptides detected, including the identification of two posttranslational modification sites. Cleavage of peptide bonds inside all seven transmembrane domains took place, generating peptides of sizes amenable to MS/MS to determine possible sequence errors or modifications within these domains. Cleavage specificity, such as glycine residue cleavage, was observed. Terminal peptides were found to be present in relatively high abundance in the hydrolysate, particularly when low concentrations of proteins were used for MAAH. It was shown that these peptides could still be detected from MAAH of bacteriorhodopsin at a protein concentration of 1 ng/microl or 37 fmol/microl. To evaluate the general applicability of this method, it was applied to identify proteins from a membrane protein enriched fraction of cell lysates of human breast cancer cell line MCF7. With one-dimensional LC-MALDI MS/MS, a total of 119 proteins, including 41 membrane-associated or membrane proteins containing one to 12 transmembrane domains, were identified by MS/MS database searching based on matches of at least two peptides to a protein. PMID- 15792717 TI - Optimization of a MALDI TOF-TOF mass spectrometer for intact protein analysis. AB - A MALDI TOF-TOF instrument was optimized and evaluated for intact protein analysis by tandem mass spectrometry. Ion source voltages and delay times were adjusted to affect an up to a 10-fold improvement in fragment ion yield compared to data obtained using default settings employed in peptide analysis. For large peptides (3-4.5 kDa), up to 90% of all possible b- and y-fragment ions were observed, which provides sufficient information for de novo sequencing and unambiguous protein identification. Product ion signals associated with preferential cleavages C-terminal to aspartic acid and glutamic acid residues and N-terminal to proline residues became dominant with increased protein molecular weight. Matrix effects were also evaluated and, among the eight matrices examined, alpha-cyano-4-hydroxycinnamic acid (CHCA) was found to produce the best intact protein tandem mass spectra for proteins up to 12 kDa. Optimized performance yielded detection limits of 50-125 fmol for proteins of 4 and 12 kDa, respectively. This improved performance has yielded an instrument with potential to be a useful tool in proteomic investigations via analysis of intact proteins. PMID- 15792718 TI - Structural characterization of cardiolipin by tandem quadrupole and multiple stage quadrupole ion-trap mass spectrometry with electrospray ionization. AB - We report negative-ion electrospray tandem mass spectrometric methods for structural characterization of cardiolipin (CL), a four-acyl-chain phospholipid containing two distinct phosphatidyl moieties, of which structural assignment of the fatty acid residues attached to the glycerol backbones performed by low energy CAD tandem mass spectrometry has not been previously described. The low energy MS2-spectra of the [M - H]- and [M - 2H]2- ions obtained with ion-trap or with tandem quadrupole instrument combined with ion-trap MS3-spectra or with source CAD product-ion spectra provide complete structural information for CL characterization. The MS2-spectra of the [M - H]- ions contain two sets of prominent fragment ions that comprise a phosphatidic acid, a dehydrated phosphatidylglycerol, and a (phosphatidic acid + 136) anion. The substantial differences in the abundances of the two distinct phosphatidic anions observed in the MS2-spectra of the [M -H]- ions lead to the assignment of the phosphatidyl moieties attached to the 1' or 3' position of central glycerol. Upon further collisional dissociation, the MS3-spectra of the phosphatidic anions provide information to identify the fatty acyl substituents and their position in the glycerol backbone. The MS2-spectra of the [M - 2H]2- ions obtained with TSQ or ITMS contain complementary information to confirm structural assignment. The applications of the above methods in the differentiation of cardiolipin isomers and in the identification of complex cardiolipin species consisting of multiple molecular structures are also demonstrated. PMID- 15792720 TI - Fragmentations and reactions of the organophosphate insecticide Diazinon and its oxygen analog Diazoxon studied by electrospray ionization ion trap mass spectrometry. AB - The fragmentations and reactions of Diazinon and related compounds have been studied by electrospray ionization ion trap mass spectrometry. Several novel fragmentation and rearrangements have been observed, including an intramolecular thiono-thiolo rearrangement. The stability, in the gas-phase, of the protomers of 2-isopropyl-4-methyl-6-pyrimidinol has been demonstrated. The complexity of the gas phase ion processes observed suggest that, at present, caution should be exercised in using this approach for the analysis of environmental and other samples until our understanding of these processes increases considerably. PMID- 15792719 TI - Statistical evaluation of electrospray tandem mass spectra for optimized peptide fragmentation. AB - Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has become the method of choice for the analysis of complex peptide mixtures. It combines the separation power of nanoflow LC with highly specific sequence analysis, allowing automated peptide sequencing with high resolution and throughput. For peptide fragmentation, the current experimental setup uses predefined parameters based on the mass-to-charge ratio of the individual precursor. Suitable parameters are typically established by empirical evaluation of fragment spectra of individual peptides used as standards. As a result, nonoptimal fragment spectra are obtained if peptides show fragmentation behavior different from these standards, which often result in the loss of sequence-specific fragment ion information. Here we describe a statistical approach for the systematic evaluation of the quality of individual peptide fragment spectra based on the calculation of their arithmetic mean and standard deviation. The method utilizes the dependence of these parameters on the difference in electric potential across the collision cell to determine the value that results in maximum information content. We show that the method is applicable to fragment spectra generated from a variety of multiply charged tryptic peptides, over a wide concentration range, and on different types of mass analyzers. We also show how this novel approach can be used to define optimized collision energy settings over a wide mass-to-charge range. PMID- 15792723 TI - Solvent-free MALDI-MS for the analysis of biological samples via a mini-ball mill approach. AB - An efficient, low sample load mini-ball mill (MBM) sample preparation procedure was developed for solvent-free MALDI analysis of peptides and proteins. Picomole sample amounts can be handled conveniently, with 30 s grinding times being sufficient. Matrix purity and molar analyte/matrix ratios are not as critical as with methods employing solvent. Ammonium salt is employed for protonation of the peptide and suppression of sodiation. This strategy allows for peptide mapping and other biochemical manipulations to be performed prior to MBM sample preparation and mass analysis. The analysis of bovine serum albumin (66 kDa) yielded good results, indicating that higher molecular weight proteins are accessible. A semi-solvent-free strategy by the MBM sample preparation method is also described. PMID- 15792724 TI - Characterization of an N-acylated glucagon-like peptide-1 derivative by electron capture dissociation. AB - An N-acylated glucagon-like peptide 1 derivative was characterized by Fourier transform ion cyclotron resonance mass spectrometry. Both electron capture dissociation (ECD) and sustained off-resonance irradiation collisionally activated dissociation (SORI-CAD) were employed. While ECD revealed full sequence coverage, site of modification, branching point, structure of the palmitoylated modification, SORI-CAD produced less complete and more ambiguous information attributable to facile losses of the fatty acid group from both parent and fragments. Thus, ECD showed a superior characterization performance over SORI-CAD in analysis of N-acylated polypeptides. PMID- 15792725 TI - Elucidation of the presence and location of t-Boc protecting groups in amines and dipeptides using on-column H/D exchange HPLC/ESI/MS. AB - High performance liquid chromatography/mass spectrometry (HPLC/MS) has become a widely used technique for routine analysis of pharmaceutical compounds. The constant search for new drugs requires the development of time-efficient methods that can be employed in high-throughput screening of combinatorial libraries of a variety of compounds, including amines and peptides. Conventional HPLC/MS is a powerful technique that can easily be automated and is suitable for comprehensive screening purposes. However, the unequivocal determination of the presence and location of important carbamoyl protecting groups of amines is often elusive because of their inherent instability under MS conditions. In this study, the use of on-column H/D exchange HPLC/ESI/MS for structure elucidation of t-Boc protecting groups which can often not be detected by MS because of facile McLafferty rearrangement has been examined. We demonstrate that employing a deuterated mobile phase in HPLC/MS analysis provides a convenient tool for the determination of the absence or presence of t-Boc protecting groups in amines and peptides. PMID- 15792726 TI - In situ formation of C-glycosides during electrospray ionization tandem mass spectrometry of a series of synthetic amphiphilic cholesteryl polyethoxy neoglycolipids containing N-acetyl-D-glucosamine. AB - In this communication, the structural analysis of six synthetic O-Linked amphiphilic cholesteryl polyethoxy neoglycolipids containing N-acetyl-D glucosamine was performed by electrospray ionization mass spectrometry in the positive ion mode, with a QqTOF-MS/MS hybrid instrument. The MS/MS analyses provided evidence for the "in situ" formation, in the collision cell of the tandem mass spectrometer, of an unexpected and unique [C-glycoside]+ product ion, resulting from an ion-molecule reaction between the N-acetyl-D-glucosamine oxonium ion and the neutral cholesta-3,5-diene molecule. Quasi MS3 analysis of this ion resulted in the dissociation of the precursor [C-glycoside]+ ion, which produced the expected third generation N-acetyl-D-glucosamine oxonium and the protonated cholesta-3,5-diene product ions. PMID- 15792727 TI - Screening and sequencing of complex sialylated and sulfated glycosphingolipid mixtures by negative ion electrospray Fourier transform ion cyclotron resonance mass spectrometry. AB - A protocol for negative ion nanoelectrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (-)nanoESI-FTICR MS, investigation of complex biological mixtures consisting of sialylated or sulfated glycosphingolipids (GSL) expressing high heterogeneity in the ceramide portion is described. Different instrumental and solvent conditions were explored and optimized to promote efficient ionization, reduce the in-source fragmentation and consequently enhance the detection of intact molecular species from complex mixtures. Using the novel optimized (-)nanoESI-FTICR MS protocol, a reliable and detailed compositional fingerprint of the polysialylated ganglioside mixture isolated from human brain was obtained. Sustained off-resonance irradiation collision-induced dissociation mass spectrometry (SORI-CID MS2) was introduced for the first time for structural elucidation of polysialylated gangliosides. Under well-defined conditions, an informative fragmentation pattern of the trisialylated ganglioside GT1 was obtained. The compositional mapping of a complex mixture of sulfated glucuronic acid containing neolacto-series GSLs extracted from bovine Cauda equina provided hard evidence upon previously described components and new structures not identified before by any other analytical method. Negative ion nanoESI-FTICR MS at 9.4 T is shown here to represent a valuable method in glycolipidomics, allowing a high resolution and mass accuracy detection of major and minor GSL glycoforms and identification of known and novel biologically relevant structures. PMID- 15792728 TI - Microwave-assisted enzyme-catalyzed reactions in various solvent systems. AB - The work describes the accelerated enzymatic digestion of several proteins in various solvent systems under microwave irradiation. The tryptic fragments of the proteins were analyzed by matrix-assisted laser desorption/ionization mass spectrometry. Under the influence of rapid microwave heating, these enzymatic reactions can proceed in a solvent such as chloroform, which, under traditional digestion conditions, renders the enzyme inactive. The digestion efficiencies and sequence coverages were increased when the trypsin digestions occurred in acetonitrile-, methanol- and chloroform-containing solutions that were heated under microwave irradiation for 10 min using a commercial microwave applicator. The percentage of the protein digested under microwave irradiation increased with the relative acetonitrile content, but decreased as the methanol content was increased. These observations suggest that acetonitrile does not deactivate the enzyme during the irradiation period; in contrast, methanol does deactivate it. In all cases, the digestion efficiencies under microwave irradiation exceed those under conventional conditions. PMID- 15792729 TI - Binding of gaseous Fe(III)-heme cation to model biological molecules: direct association and ligand transfer reactions. AB - The binding of a variety of ligands with Fe(III)-heme(+) ion, prosthetic group of heme proteins, has been studied in the gas phase by ESI-FT-ICR mass spectrometry. The ligands have been selected among substrate molecules of heme proteins (e.g., NO, nitroso compounds) or among model compounds acting for the functional groups that are present in the protein backbone (e.g., amines, thioethers, nitriles, ketones, amides, etc.). Both the kinetic and the thermodynamic features of the addition reactions are reported. Fe(III)-heme(+) ions react faster with lone pair donor ligands as the reaction becomes increasingly thermodynamically favored (higher heme cation basicity of the ligand, HCB, namely -DeltaG degrees for the ligand addition reaction). In turn HCBs correlate in general with the gas phase basicity toward the proton of the various ligands. A ligand addition equilibrium is established with weaker ligands, methanol, acetonitrile and acetone, yielding absolute HCB values, whereas ligand transfer equilibriums allowed a scale of relative (and absolute) HCBs to be constructed. NO displays exceptional binding properties towards Fe(III)-heme(+), unrelated to the low gas phase basicity toward the proton of this molecule, which is clearly the basis for the paramount role of heme proteins in NO binding and regulation. PMID- 15792730 TI - Combination antibiotic therapy for Pseudomonas aeruginosa bacteraemia. PMID- 15792732 TI - Mass antibiotics for trachoma and the Allee effect. PMID- 15792733 TI - Visceral leishmaniasis--an opportunistic infection in HIV-infected patients. PMID- 15792738 TI - Mechanisms of action of newer antibiotics for Gram-positive pathogens. AB - Certain Gram-positive bacteria, including meticillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and quinolone-resistant Streptococcus pneumoniae have achieved the status of "superbugs", in that there are few or no antibiotics available for therapy against these pathogens. Only a few classes of novel antibiotics have been introduced in the past 40 years, and all since 1999, including the streptogramin combination quinupristin/dalfopristin (Synercid), the oxazolidinone linezolid, and the lipopeptide daptomycin. This review discusses the mechanisms of antibiotic action against Gram-positive pathogens, and resistance counter-mechanisms developed by Gram-positive bacteria, with emphasis on the newer agents. PMID- 15792739 TI - Natural history and pathophysiology of Q fever. AB - Q fever is a zoonosis caused by Coxiella burnetii. Infection with C burnetii can be acute or chronic, and exhibits a wide spectrum of clinical manifestations. The extreme infectivity of the bacterium results in large outbreaks and makes it a potential bioweapon. In the past decade, the complete genome sequencing of C burnetii, the exploration of bacterial interactions with the host, and the description of the natural history of the disease in human beings and in experimental models have all added to our knowledge about this fascinating disease. Advances in understanding the pathophysiology and natural history of Q fever are reviewed. PMID- 15792741 TI - The challenges were legion. AB - The search for the cause of the large outbreak of severe pneumonia that affected attendees of a convention of war veterans in Philadelphia in 1976 was complicated by a number of factors. The 2-10 day incubation period meant that cases were dispersed across Pennsylvania at time of onset, and matters were further complicated by the lack of a centralised record of attendance of the convention. Known methods for culture, serology, tissue staining, and toxicology gave negative results. The reliance on a clinical case definition, and the need to coordinate local, state, and federal jurisdictions both hindered efforts. Intense public scrutiny, anxiety, and promotion of implausible theories, researchers' memories of prior unsolved major investigations, and the inherent difficulty of documenting airborne spread also had an impact. The novel bacterial agent Legionella pneumophila was identified and its spread defined by persistent application of basic epidemiological and laboratory principles, including the willingness to re-examine one's assumptions. PMID- 15792740 TI - Hepatitis C virus, cryoglobulinaemia, and vasculitis: immune complex relations. AB - Several viruses are involved in the development of systemic vasculitides. Hepatitis C virus (HCV) has been shown to be closely related to mixed cryoglobulinaemia, an immune complex-mediated vasculitis. HCV particles and non enveloped nucleocapsid protein participate in the formation of immune complexes. Once formed, immune complexes precipitate in many organs, including the skin, kidneys, and peripheral nerve fibres. Viral proteins confer peculiar physical and chemical properties on cryoimmunoglobulins. Since expansion of rheumatoid factor synthesising B cells is the biological hallmark of mixed cryoglobulinaemia, it may be that the combination of rheumatoid factor activity and cryoprecipitability is responsible for the vasculitis. B-cell clonal expansion occurs primarily in the liver and correlates with a high intrahepatic viral load, pointing to a major role for HCV in the emergence and maintenance of B-cell clonalities. Recognition of HCV as an aetiological factor in most cryoglobulinaemic vasculitides has dramatically changed the approach to their treatment. Emphasis, in fact, is now placed on abatement of the viral load and deletion of B-cell clonalities. PMID- 15792742 TI - Considerations for a WHO European strategy on health-care-associated infection, surveillance, and control. AB - Health-care-associated infection (HAI) is a major issue of patient safety with a substantial impact on morbidity, mortality, and use of additional resources worldwide. In April 2004, the WHO Regional Office for Europe organised the first international consultation to address the issue of HAI in eastern and central Europe. The main objectives of the consultation were to identify the primary needs and obstacles for the prevention and control of HAI at country level, to design the essential components of an international strategy to effectively address the issue of HAI, and to identify specific priorities and recommendations for interventions by the WHO and other international institutions. An update on HAI activities and related networks throughout Europe, together with the outcome of the meeting, are presented, with special emphasis on future considerations for a European WHO strategy on HAI prevention. PMID- 15792743 TI - Thoracic aortic graft infection after disseminated tuberculosis. PMID- 15792744 TI - Fever, backache, and groin pain. PMID- 15792745 TI - On the quest for Six Sigma. AB - A review of patient safety from a surgical perspective with emphasis on erosion of hierarchy, human factors, and an institutional implementation of multidisciplinary team training to create highly effective dynamic teams. Suggestions include further opportunities to enhance patient safety in surgical patients. PMID- 15792746 TI - On the uniqueness of surgery. AB - BACKGROUND: As the divergence between medicine and surgery increases, the author traces its beginnings not to the recent past, but as far back as the Middle Ages and the Renaissance. Even at that time, physicians rarely came in physical contact with the patients and made their diagnoses from looking at flasks of urine from the patients. Skilled physicians allegedly could make approximately 1000 diagnoses from examining a flask of urine. On the other hand, barber surgeons were relatively unskilled and unrewarded, at a much lower class in society, and laid hands on the patient by draining abscesses, cauterizing, dealing with bladder stones, etc. METHODS: Despite the Quincentenary of the Royal College of Surgeons of Edinburgh this year and the rendering of the first guild to the barber surgeons, this schism between medical and surgical practice has remained. The author attempts to fit this schism into medicine today by addressing continuity of care and the 80-hour workweek, stating that this schism is not a new development but has existed for many years. CONCLUSIONS: The proposal is made that medicine and surgery should work together as much as they can. However, if surgical patients are not well served by the lack of emphasis on continuity of care brought forth by the leadership of medicine in this country, who the author claims of intentionally distancing themselves from patient care, then surgery may need to go its own way. The patient comes first. PMID- 15792747 TI - Special address to the Midwest Surgical Association on surgical outcome analysis and patient safety. PMID- 15792749 TI - Outcome of severe acute pancreatitis. AB - BACKGROUND: The treatment of severe acute pancreatitis has been evolving from routine operative management to nonoperative care for patients without evidence of pancreatic infection. METHODS: Retrospective chart review of patients with severe acute pancreatitis at a single institution during a 9-year period. RESULTS: Sixty consecutive patients had severe pancreatitis. Forty-two had pancreatic necrosis on computed axial tomography (13 infected and 29 sterile). Patients with infected necrosis and 8 with sterile necrosis had operative debridement; the remaining patients were managed without operation (n = 39). The overall mortality was 15%. Mortality was directly related to the Acute Physiology and Chronic Health Examination II and Marshall organ failure scores (P <0.001). Patients who died had a greater incidence of nosocomial infection. CONCLUSIONS: Patients with infected pancreatic necrosis require early operative debridement, whereas those with sterile necrosis or severe pancreatitis without necrosis can usually be managed safely without surgery. PMID- 15792748 TI - Randomized controlled trial comparing the controlled rehabilitation with early ambulation and diet pathway versus the controlled rehabilitation with early ambulation and diet with preemptive epidural anesthesia/analgesia after laparotomy and intestinal resection. AB - BACKGROUND: Multimodal postoperative care regimens accelerate recovery after abdominal surgery. The benefit of thoracic epidural (TE) analgesia over patient controlled analgesia (PCA) remains unproven when used with a fast-track postoperative care plan. METHODS: Fifty-six patients undergoing major intestinal resection, and on a fast-track postoperative care plan, were randomized to preemptive TE or PCA. Patients were evaluated at standard time points for pain score, quality of life (Short Form-36), and complications. Oral analgesia was substituted for TE and PCA on the second postoperative day. Discharge criteria were identical for both groups. RESULTS: Six patients (20.6%) had a failed epidural. There was no difference in length of stay (5.8 versus 6.2 days, TE versus PCA, P = .55), total length of stay (including readmissions), pain scores, quality of life, complications, or hospital costs at any time point. CONCLUSION: TE offers no advantage over PCA for patients undergoing major intestinal resections who are on a fast-track postoperative care plan using PCA. PMID- 15792750 TI - Increased preoperative platelet count is associated with decreased survival after resection for adenocarcinoma of the pancreas. AB - BACKGROUND: Platelets are thought to participate in tumor metastasis. However, the relationship between platelet count and prognosis in pancreatic cancer remains unresolved. METHODS: A chart review of patients undergoing resection for pancreatic adenocarcinoma was undertaken. Demographic, perioperative, and outcome data were collected. Kaplan-Meier survival and Cox regression analyses were used to determine the impact of preoperative platelet count on survival. RESULTS: Between June 1995 and March 2003, 109 patients (63% male) with a median age of 68 years (range 42 to 85 years) underwent resection for pancreatic cancer. Univariate analysis demonstrated that platelet count, lymph node or margin status, and histology were associated with survival. In multivariate analysis, the association between increased platelet count and poor survival maintained significance. CONCLUSIONS: Increased preoperative platelet count is associated with adverse survival outcome in patients undergoing resection for pancreatic cancer. Antiplatelet medications warrant further study in an attempt to improve survival in these patients. PMID- 15792751 TI - Inguinal neurectomy for inguinal nerve entrapment: an experience with 100 patients. AB - BACKGROUND: Inguinal nerve entrapment is a debilitating postoperative problem. PATIENTS AND METHODS: One hundred patients were treated for inguinal nerve entrapment, including 52 men and 48 women with an average age of 45 +/- 14 years. Most patients had inguinal hernia repairs or Pfannenstiel incisions. Mesh was found in 27% of patients. Symptoms included pain (100%), radiation of pain to the thigh and/or genital area (59%), and postural pain (59%). Diagnosis was made by physical examination, postural maneuvering, and inguinal nerve block. Proximal nerve resection was followed by Mersiline (Ethicon, Inc., Somerville, NJ) ligature and absolute alcohol or phenol application to prevent neuromas. RESULTS: Five percent of patients had minor complications. There was abnormal nerve histopathology in 18%. Total pain relief was attained in 72% of patients, partial relief in 25%, and no relief in 3%. Two patients complained of numbness postoperatively. Multifactorial analysis showed recurrent hernia repair as a significant predictive factor. PMID- 15792752 TI - Assessing the competencies in general surgery residency training. AB - BACKGROUND: Accreditation Council for Graduate Medical Education (ACGME) competencies have increased the focus on education. A 1-year study of observational assessments was conducted. METHODS: "Point of Observation" evaluations were completed by faculty for postgraduate year (PGY) I and II surgery residents. Resident procedures and patient office visits were rated in 9 categories, using a Likert scale. Interns were expected to perform at a novice level (0-30%) and PGY II residents at an advanced level, with improvements expected. RESULTS: PGY I and II residents showed overall improved scores (12% and 6%, respectively) over 1 year. Intern improvements in 9 categories were linear, ranging from 0% to 48%. PGY II resident scores were more variable, with improvements noted in 6 categories ranging from 10% to 30%. Three categories declined in scores, ranging from 2% to 18%. CONCLUSIONS: Competencies bring emphasis to education and measurable outcomes. Early efforts have proven valuable in identifying curriculum and learning needs. PMID- 15792753 TI - Early tracheostomy versus late tracheostomy in the surgical intensive care unit. AB - BACKGROUND: This study's purpose was to determine if early tracheostomy (ET) of severely injured patients reduces days of ventilatory support, the frequency of ventilator-associated pneumonia (VAP), and surgical intensive care unit (SICU) length of stay (LOS). METHODS: This 2-year retrospective review included 185 SICU patients with acute injuries requiring mechanical ventilation and tracheostomy. ET was defined as 7 days or less, and late tracheostomy (LT) as more than 7 days. RESULTS: The incidence of VAP was significantly higher in the LT group, relative to the ET group (42.3% vs. 27.2%, respectively; P <.05). Acute Physiology and Chronic Health Evaluation II scores, hospital and SICU LOS, and the number of ventilator days were significantly higher in the LT group. CONCLUSIONS: In patients who required prolonged mechanical ventilation, there was significant decreased incidence of VAP, less ventilator time, and lower ICU LOS when tracheostomy was performed within 7 days after admission to the SICU. PMID- 15792754 TI - Lysyl oxidase expression in bronchogenic carcinoma. AB - BACKGROUND: Lysyl oxidase catalyzes a key step in the cross-linking of collagen and elastin in the extracellular matrix. Recent studies have documented differential lysyl oxidase expression in the stromal reaction to colon, breast, prostate, and lung cancer. The present study was undertaken to test the hypothesis that lysyl oxidase mRNA and protein expression decrease with advancing tumor stage in patients with bronchogenic carcinoma. METHODS: Tumor specimens were obtained from 17 patients undergoing resection for bronchogenic carcinoma. Real-time polymerase chain reaction was used to determine steady-state lysyl oxidase mRNA expression, and protein expression was qualitatively assessed by immunohistochemistry. RESULTS: Real-time polymerase chain reaction studies documented a 3.4-fold graded decrease in lysyl oxidase mRNA levels as tumors progressed from stage I to IV. Similar qualitative changes in lysyl oxidase protein expression were demonstrated by immunohistochemistry. CONCLUSIONS: These results support the hypothesis that variations in lysyl oxidase expression may correlate with the invasive and metastatic potential of bronchogenic carcinoma. PMID- 15792755 TI - Killing tumor cells: the effect of photodynamic therapy using mono-L-aspartyl chlorine and NS-398. AB - BACKGROUND: Photodynamic therapy (PDT) is a useful treatment for malignant tumors. PDT involves the administration of a photosensitive drug that is selected by neoplastic tissues and their vasculature. One such photosensitizer is mono-l aspartyl chlorine e6 (NPe6). Recent evidence suggests that the presence of the cyclooxygenase-2 (COX-2) inhibitor NS-398 may potentiate the effect of photosensitizing agents. This study was designed to determine if the addition of NS-398 to NPe6-induced PDT in single or fractionated dosing would result in greater tumor kill. METHODS: Colon-38 tumor was subcutaneously implanted into both flanks of mice and allowed to grow to 0.5 to 1.0 cm. Mice were randomly allocated to 5 groups: (1) single dose of NPe6; (2) fractionated dose of NPe6; (3) NS-398 only; (4) single dose of NPe6 + NS-398; and (5) fractionated dose of NPe6 + NS-398. The left flank was shielded from exposure to irradiation. Tumor size was measured before initiation of PDT and at the time of sacrifice. RESULTS: The initial tumor weights of both flanks were not significantly different between all groups. Tumor weights at the time of death after PDT using NPe6 were significantly less than their paired tumors in the untreated flanks (P <0.0001). Tumor weights in the treated flanks were significantly less in the group receiving the fractionated dosing of NPe6 as compared to the single dose of NPe6 (P = 0.0037). NS-398 plus the single dose of NPe6 significantly decreased tumor weight in the PDT-treated flank (P = 0.035) at a level equivalent to that observed with fractionated dosing of the photosensitizer in the absence of NS 398. NS-398 did not significantly further decrease tumor weight in the group that received the fractionated dose of NPe6. CONCLUSIONS: Fractionated dosing of NPe6 demonstrated the best tumor kill. However, NS-398 did not potentiate the effect of PDT using fractionated dosing of NPe6. While PDT using the single NPe6 dose significantly decreased tumor weight, the addition of NS-398 potentiated the killing effect. PMID- 15792756 TI - Intraoperative parathyroid hormone levels in thyroid surgery are predictive of postoperative hypoparathyroidism and need for vitamin D supplementation. AB - BACKGROUND: Intraoperative parathyroid hormone (ioPTH) levels are not monitored routinely in thyroid surgery, although they are used widely during parathyroidectomy as an indicator of parathyroid gland function. This prospective study evaluated the occurrence of hypoparathyroidism after thyroid surgery and the use of ioPTH levels to predict the need for postoperative vitamin D supplementation. METHODS: Seventy-two patients underwent thyroidectomy or neck dissection by 1 surgeon. Forty-five patients had a total thyroidectomy, 16 patients had a hemithyroidectomy, 9 patients had a completion thyroidectomy, and 2 patients had a neck dissection alone for recurrent thyroid cancer. ioPTH and serum calcium (SCa) levels were obtained during the course of surgery and 1 month after surgery. Levels from these time points were compared, and correlated with the need for vitamin D supplementation at the 1-month follow-up evaluation using the Fisher exact test. RESULTS: Of the 72 patients, 14 had an ioPTH level less than 10 pg/mL at closure. At the 1-month evaluation, 11 of these 14 patients required vitamin D supplementation because of persistent hypoparathyroidism or hypocalcemia (P <.001). The remaining 3 of the 14 patients with ioPTH levels less than 10 pg/mL at closure did not require vitamin D supplementation at the 1-month evaluation because they were asymptomatic and their PTH and SCa levels had normalized. None of the 58 patients with an ioPTH level greater than 10 pg/mL at closure needed vitamin D supplementation at the 1-month follow-up evaluation. CONCLUSIONS: An ioPTH level less than 10 pg/mL at closure is a strong predictor of hypoparathyroidism after thyroid surgery. Patients with ioPTH levels less than 10 pg/mL at closure should be placed on vitamin D supplementation after surgery to anticipate decreased parathyroid gland function and to avoid symptomatic hypocalcemia. PMID- 15792757 TI - A comparison of corrected serum calcium levels to ionized calcium levels among critically ill surgical patients. AB - BACKGROUND: Aberrations in calcium homeostasis are common in critically ill patients. The proper method to evaluate this issue in surgical patients has not been completely defined. METHODS: Medical records of patients admitted to a university-affiliated, tertiary-care surgical intensive care unit were retrospectively reviewed. Calcium status was evaluated by ionized levels and as a function of serum calcium levels corrected for albumin aberrations. RESULTS: Corrected serum calcium values failed to accurately classify calcium status in 38% of cases. The sensitivity and specificity of the corrected serum calcium formula to evaluate hypocalcemia were 53% and 85%, respectively. Corrected serum values underestimated the prevalence of hypocalcemia and overestimated the prevalence of normocalcemia. No factors were able to discern which patients could be evaluated by corrected serum calcium levels. CONCLUSIONS: Calcium homeostasis should be evaluated by ionized calcium levels rather than as a function of serum calcium and albumin levels. PMID- 15792758 TI - Reinforcement of subcuticular continuous suture closure with surgical adhesive strips and gum mastic: Is there any additional strength provided? AB - BACKGROUND: This study aimed to compare the burst strength of suture closure versus the use of suture and strip together. METHODS: On cadavers, 50 skin incisions were closed as follows: group 1--subcuticular continuous suture; group 2--same suturing with placement of strips; group 3--same as group 2 except gum mastic was applied prior to strips; group 4--strips alone; and group 5--strips with gum mastic application. The separation forces were measured using a tensilometer. RESULTS: The mean separation forces were as follows: group 1, 14.17 kg; group 2, 14.37 kg; group 3, 15.39 kg; group 4, 1.52 kg; and group 5, 3.85 kg. There were no statistically significant differences between groups 1, 2, and 3. When compared with group 4, group 5 required markedly more force to separate the wound. CONCLUSIONS: Strip reinforcement with/without gum mastic did not provide any additional strength when sutures were used. Gum mastic increased the adherence of strips and this was important when strips were the only means of wound closure. PMID- 15792759 TI - Influence of fibrin glue on seroma formation after breast surgery. AB - BACKGROUND: This study was designed to determine the effectiveness of Hemaseel APR fibrin sealant versus conventional drain placement in the prevention of seromas after breast procedures. METHODS: A prospective, randomized, controlled study of subjects who were randomized into control (drain) and experimental (fibrin) groups was conducted. RESULTS: Analysis of 82 patients showed similarly matched groups. Seroma formation rate was 45.5% in the control group and 36.8% in the fibrin glue group (P = 0.43). The rate of wound complications was similar. Aspirate volumes were significantly greater in the fibrin glue group. Drain placement saved patients >366 US dollars over fibrin glue. CONCLUSIONS: Although use of fibrin sealant resulted in a nonsignificant decrease in seroma formation rate compared with that of drain placement, the higher cost involved, cumbersome technique, and higher aspirate volumes tend to indicate that there is no advantage to using fibrin glue over drain placement with the technique described. PMID- 15792760 TI - Establishing transfusion needs in burn patients. AB - BACKGROUND: Despite blood-conservation techniques, hemorrhage during burn excision remains substantial. It is difficult to predict the blood loss that will occur per operation and how many units the patient will require during surgery. This may result in high cross-match-to-transfusion ratios (CMTRs). METHODS: A retrospective chart review from 2001 to 2003 was performed. All adult patients with >20% total body surface area burns who underwent surgery were included in the study. Variables examined were centimeters excised, estimated blood loss, packed red blood cells transfused during surgery, preoperative and postoperative hematocrit, and CMTR. RESULTS: There were 273 operations. The average estimated blood loss was 820 mL, and; the median posttransfusion hematocrit was 27. Based on the area excised and units transfused, a ratio of packed red blood cells/cm excised was determined. A total of 1.78 U blood were transfused/1000 cm(2) excised to keep hematocrit between 25 and 31; P = 0.02. CONCLUSIONS: Estimation of excision area can predict transfusion need, which at our institution yields a low CMTR. PMID- 15792761 TI - Epidural versus intravenous pain control in elderly patients with rib fractures. AB - BACKGROUND: Epidural catheters are used in older patients with rib fractures to improve outcome. We reviewed the efficacy of epidural analgesia (EA) compared with intravenous narcotics (IVN) in this population. METHODS: Rib fracture patients >55 years old admitted to our level I trauma center from 1999 through 2002 were reviewed for demographics, Injury Severity Score (ISS), Abbreviated Injury Score for chest, length of stay, cardiopulmonary comorbidities, complications, and type of analgesia. RESULTS: There were 187 patients: 72 men and 115 women. The mean age was 77 years. For ISS <9, length of stay for EA patients was 12 +/- 5 days versus 5 +/- 4 days for IVN patients (P < 0.001). Complications occurred in 9 of 10 EA patients versus 21 of 52 IVN patients (P < 0.001). No difference was noted in length of stay for patients with ISS > or =9. Complications in the high ISS group occurred in 29 of 43 EA patients versus 37 of 82 IVN patients (P <0.05). Stratification of patients based on low versus high Abbreviated Injury Score for chest yielded similar results. CONCLUSIONS: EA is associated with prolonged length of stay and increased complications in elderly patients, particularly those with less significant injuries, regardless of cardiopulmonary comorbidities. EA for elderly patients with rib fractures should be prospectively re-evaluated. PMID- 15792763 TI - Complications arising from splenic embolization after blunt splenic trauma. AB - BACKGROUND: Nonoperative management (NOM) of splenic trauma is now the standard in stable trauma patients. Splenic artery embolization (SAE) is an increasingly used adjunct to NOM. We examined complications arising from SAE. METHODS: Patients admitted to a level I trauma center with splenic trauma over a 26-month period were identified. Management method, operative or nonoperative, was noted. SAE patients were analyzed in detail. RESULTS: There were 284 splenic trauma admissions. Ninety-three patients underwent operative management, and 191 received NOM. Fifteen patients (7.8%) underwent SAE. Embolization was proximal in 10, distal in 1, and combined in 4 patients. No NOM failures occurred. Major complications (27%) included splenic bleeding, splenic infarction, splenic abscess, and contrast-induced renal insufficiency. Minor complications of fever, pleural effusions, and coil migration occurred in 53% of patients. No relationship between SAE location and the presence of complications was noted. CONCLUSIONS: SAE is an effective and safe procedure. Both major and minor complications can arise after SAE. PMID- 15792762 TI - Recombinant factor VIIa for the treatment of severe postoperative and traumatic hemorrhage. AB - BACKGROUND: The aim of this study was to determine the dose of recombinant factor VIIa (rFVIIa) that has been used in our institution to successfully control hemorrhage in trauma and postoperative patients. METHODS: This was an 8-month retrospective cohort study of 13 patients with acute hemorrhage and no known history of coagulopathic disorders. RESULTS: Administration of factor VIIa resulted in the cessation of life-threatening hemorrhage at dosages approximately one half those recommended for the management of hemophilia. After administration, there was a significant decrease in the total blood-product transfusion requirement (P <0.05). CONCLUSIONS: The use of factor VIIa in patients with life-threatening hemorrhage is a safe and effective therapeutic modality when used as an adjunct to standard interventions for control of severe hemorrhage. Lower-dose regimens were as successful as higher-dose regimens previously reported. The results of this respective study of 13 patients suggests that recombinant factor VIIa therapy for control of life-threatening hemorrhage as an adjunct to standard interventions can be successful at doses <90 mg/kg. PMID- 15792764 TI - Ductal lavage in the high-risk patient. AB - BACKGROUND: [corrected] This analysis was conducted in a single-surgeon clinical practice to evaluate the utility and practicality of performing ductal lavage in a population determined to be at high risk for breast cancer. METHODS: One hundred twenty patients with negative mammograms and/or negative breast examinations had nipple aspiration and ductal lavage performed by a single surgeon. All were at high risk either according to Gail risk score, a previous breast carcinoma, or nipple discharge. RESULTS: One hundred twenty patients underwent nipple aspiration. Thirty-two did not undergo lavage: 29 had no fluid, and 3 had unsuccessful cannulation. Eighty-eight underwent lavage: 15 had insufficient epithelial content, 51 had benign cytology, and 22 had abnormal cells. Of the 22, 20 had mild atypia, 1 had marked atypia, and 1 had malignant cells. CONCLUSIONS: Ductal lavage can be done in a surgical practice and can stratify risk for the individual patient. This is important to both patient and surgeon in formulating a treatment plan based on objective cytologic criteria. PMID- 15792765 TI - Is operative management effective in treatment of perforated typhoid? AB - BACKGROUND: Salmonella typhi infection continues to be a significant problem worldwide. Patients suffering from "typhoid" in endemic regions such as West Africa often present late in the course of the disease with symptoms of malnutrition and peritonitis. Clinical peritonitis in these patients is invariably associated with perforation of the terminal ileum and purulent peritonitis. Operative intervention and its success have not been well documented. The purpose of this article is to review the experience of 1 hospital in West Africa and the efficacy of operative management of perforated intestine from typhoid. METHODS: A retrospective review of all patients admitted between January and October of 2003 to the Carolyn Kempton Memorial Hospital in Togo, West Africa, with the diagnosis of typhoid. Demographic data including age, sex, number of perforations, and outcome were obtained. All patients were treated with ampicillin, gentamycin, and flagyl or chloramphenicol as the sole antibiotic therapy along with operative management. RESULTS: One hundred ninety-one patients with typhoid were admitted. One hundred twelve patients underwent laparotomy for perforation, 18 of whom died from persistent typhoid septicemia for a mortality rate of 16%. Reoperative management was used in some patients who did not respond immediately. CONCLUSIONS: Primary repair of typhoid perforation is a safe and effective treatment. Patients with persistent septicemia after laparotomy most likely have resistant Salmonella typhi infection. Availability of second-line antibiotic therapy in these third world countries would likely improve outcomes. Prospective studies on appropriate antibiotic therapy along with operative management in endemic areas are necessary until resources are available for preventative measures. PMID- 15792766 TI - Spontaneous retroperitoneal hemorrhage. AB - BACKGROUND: We evaluated patients with spontaneous retroperitoneal hemorrhage for reliable predictors of early diagnosis and improved outcomes. METHODS: A retrospective chart review was done to determine patient demographic and laboratory findings, presenting symptoms, time to diagnosis, anticoagulant and/or antiplatelet agent use, transfusions, and patient outcome. RESULTS: One hundred nineteen patients were identified; 14 (12%) died (mean age 77 +/- 9 years vs. 74 +/- 10 years for survivors [P = 0.235]). All nonsurvivors were on anticoagulants: 8 of 89 (9%) were on heparin or warfarin alone, and 6 of 23 (26% [P = 0.028]) were on a combined anticoagulant-antiplatelet regimen. Symptom onset to computed axial tomography (CAT) scan averaged 1.3 +/- 1.3 days for nonsurvivors versus 1.5 +/- 1.9 days for survivors (P = 0.778). Hemoglobin was 9.07 +/- 3.35 for nonsurvivors versus 9.60 +/- 2.07 for survivors (P = 0.435). Eighty-eight patients were transfused, and 10 died; 31 patients had no transfusion, and 4 of these died (P = 0.821). CONCLUSIONS: A high index of clinical suspicion is necessary for diagnosis of spontaneous retroperitoneal hemorrhage because these patients present with a variety of symptoms. Prospective studies are necessary to determine whether earlier diagnosis combined with aggressive resuscitation can impact the high mortality rate seen in these patients. PMID- 15792767 TI - The era of ultrasonography during laparoscopic cholecystectomy. AB - BACKGROUND: The use of ultrasound cholangiography during cholecystectomy has been well described. This study was undertaken to assess the use of the umbilical port exclusively for ultrasound and to assess its employment on the use of fluoroscopy resources. In addition, we also looked at the increased use of ultrasound from 2000 to 2004. METHODS: The use of imaging techniques during all cholecystectomies was analyzed from January 2000 to July 2001 for one surgeon and compared with that surgeon's present use from January 2004 to June 2004. Patient demographics, intraoperative finding, and postoperative results were reviewed. RESULTS: During the first study period, ultrasound was used in 29% of 189 laparoscopic cholecystectomies. During 2004, ultrasound was used in 77% of 66 laparoscopic cholecystectomies. Throughout both periods, fluoroscopy was only used during 6 laparoscopic common bile duct explorations (2.4% of all cases). There were no false-positive or -negative ultrasounds, and there were no bile duct injuries. CONCLUSIONS: As experience with ultrasound cholangiography increases, there is little indication for fluoroscopic cholangiography except for rare questions concerning anatomy and during therapeutic maneuvers for common bile duct stones. PMID- 15792768 TI - Open Roux-en-Y gastric bypass in 925 patients without mortality. AB - BACKGROUND: Open Roux-en-Y gastric bypass has become the gold standard for bariatric surgery. Safety has always been a concern with this elective surgery, especially among the general public. With increasing numbers of bariatric surgeries being performed, the public eye is once again focused on safety and outcomes for these patients. METHODS: Nine hundred twenty-five consecutive open Roux-en-Y gastric bypass patients were reviewed. Charts were retrospectively reviewed for early complications, late complications, and resolution of medical comorbidities. RESULTS: There were no deaths in this study group. The average body mass index (BMI) was 51. Eight leaks at the anastomosis occurred with no reoperations. Hypertension resolved in 70% and diabetes mellitus resolved in 58% of patients. CONCLUSIONS: Open Roux-en-Y gastric bypass is a safe operation, even with increasing numbers of surgeries being performed. Major complications are low and improvement of medical comorbidities is significant. A multidisciplinary team approach helps to improve care and clinical outcomes. PMID- 15792769 TI - Stricture dilation after laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: In surgical treatment of morbid obesity, maintaining a restrictive anastomosis is key to long-range success. However, laparoscopic Roux-en-Y gastric bypass (LRYGB) may result in gastrojejunal (GJ) stricture, requiring treatment in up to 27% of patients. METHODS: This is a retrospective review of the outcome of 223 consecutive LRYGB patients. Patients developing stricture received standard endoscopic balloon dilation by the same surgeon. Stricture and nonstricture groups were compared for excess body weight loss (EBWL) at 1, 3, 6, and 12 months. RESULTS: GJ stricture requiring dilation occurred in 38 patients (17%). After dilation all patients were relieved of stricture symptoms and none required revision. By 12 months, patients with stricture had an EBWL of 86% compared with nonstrictured patients at 75%. CONCLUSION: Endoscopic balloon dilation is a safe and effective treatment option for GJ stricture. Improved weight loss occurred for patients with stricture requiring dilation. PMID- 15792770 TI - Safety and efficacy of superselective angioembolization in control of lower gastrointestinal hemorrhage. AB - BACKGROUND: We evaluated the safety and efficacy of angioembolization to control lower gastrointestinal hemorrhage. METHODS: Retrospective chart review of patients undergoing angiography for lower gastrointestinal hemorrhage from January 2000 to December 2002. RESULTS: Seventy-seven patients with lower gastrointestinal hemorrhage underwent mesenteric angiography. Angioembolization was performed in 11 patients. Sixty-six patients were not embolized; 47 of these were treated medically and 19 surgically. Mortality rate was not significantly different in patients treated surgically (3 of 19, 16%) versus those managed medically (6 of 47, 13%; P = 0.746). Of the 11 patients who were embolized, 10 had immediate cessation of hemorrhage, 7 had gastrointestinal ischemia, and 6 died (55%). Overall mortality in non-embolized patients was 9 of 66 (14%; P = 0.002 versus mortality in embolized patients). CONCLUSIONS: Angioembolization, though effective at controlling hemorrhage, is associated with ischemic complications and a high mortality rate. Our data support surgical or medical management for lower gastrointestinal hemorrhage. PMID- 15792771 TI - Cancer biology and necrotic changes in metastatic lymph nodes and survival of colon cancer patients. AB - BACKGROUND: Identifying factors that can contribute to a better understanding of tumor progression in stage III colon cancer patients continues to be an important task. Necrotic changes in metastatic lymph nodes have not been previously analyzed in English literature. METHODS: The study included 48 consecutive colon and rectosigmoid cancer patients with stage III disease who underwent radical surgery. After reviewing the diagnostic slides, a pathologist developed a scale describing the extent of necrotic changes. Results were evaluated using Kaplan Meier method and log-rank test. RESULTS: Thirty-four (70%) patients had necrotic changes in metastatic lymph nodes. Patients with necrotic changes in metastatic lymph nodes had more risk factors than patients without necrosis. The 5-year survival rate for patients with necrotic changes in metastatic lymph nodes was 85% and for patients without necrosis was 50% (P = 0.02). CONCLUSIONS: The survival of patients with necrotic changes in metastatic lymph nodes was higher (P = 0.02). These necrotic changes can help us to understand body-tumor relations. PMID- 15792773 TI - Retrofascial mesh repair of ventral incisional hernias. AB - BACKGROUND: Recurrence rates after ventral incisional hernia repair are reported to be as high as 33% and are associated with considerable morbidity and lost time. The purpose of this study was to determine if retrofascial mesh placement reduces the incidence of recurrence as well as the severity of wound infections. METHODS: A prospective database covering the period from January 1995 to June 2003 was maintained. All patients underwent a standardized technique by a single surgeon. Polypropylene mesh was placed between the fascia and the peritoneum with the fascia closed over the mesh. RESULTS: There were 150 patients (126 women, 24 men) with a mean age of 55 years. Their average weight was 88 kg, with an average body mass index of 32. Sixty-three (42%) of the hernias were recurrences of a previous repair. The average size of the hernia was 8 x 14 cm. There was 1 postoperative mortality. There was a 9% postoperative infection rate with 2 patients (1%) requiring mesh removal. Long-term follow-up evaluation has revealed 3 recurrences (2%) and 3 readmissions for bowel obstruction with 1 patient requiring surgical release. There were no fistulas noted. CONCLUSIONS: Incisional hernia repair with mesh placed in the retrofascial position decreases both the risk for recurrence and the severity of wound infection without significant problems from bowel obstruction or enteric fistula. PMID- 15792772 TI - Life-threatening tertiary hyperparathyroidism in the critically ill. AB - BACKGROUND: Tertiary hyperparathyroidism typically occurs in patients who have recovered from renal failure after renal transplantation. This report describes a syndrome of tertiary hyperparathyroidism after recovery from multiple organ failure (MOF) with acute oliguric renal failure (AORF). METHODS: Six patients with MOF including AORF are presented. Increased parathyroid hormone (PTH) levels were documented as early as 3 weeks after injury or septic insult and remained increased in some patients for several weeks. RESULTS: The resultant increase in calcium levels led to recurrent bouts of bradycardia, often leading to asystole requiring cardiopulmonary resuscitation. Hypercalcemic-induced bradycardia was refractory to hydration, loop diuresis, atropine, and external pacing. Definitive treatment requires bisphosphonate therapy, which must be repeated until organ function has returned to normal. CONCLUSIONS: A new syndrome of life-threatening tertiary hyperparathyroidism is described in patients with critical illness. This syndrome probably is being overlooked frequently in critical care units. Early diagnosis and prophylactic treatment with bisphosphonate may preclude the life threatening cardiac arrhythmias. PMID- 15792774 TI - Does the width of the gastric tube matter in esophageal reconstruction? PMID- 15792775 TI - Anti-proliferating effect of iptakalim, a novel KATP channel opener, in cultured rabbit pulmonary arterial smooth muscle cells. AB - ATP-sensitive potassium (K(ATP)) channels of pulmonary arterial smooth muscle cells (SMCs) have been implicated in pulmonary hypertension. Iptakalim, designed and synthesized by ourselves, is a newly selective K(ATP) channel opener. Here, we explored the effects of iptakalim on the rise of cytoplasmic free Ca(2+) concentration ([Ca(2+)](cyt)) induced by endothelin-1 (ET-1) and on the proliferation of cultured rabbit pulmonary arterial SMCs. The results showed that iptakalim inhibited the [Ca(2+)](cyt) increase. The enhanced [(3)H]thymidine incorporation was inhibited and the transition of cells from static phase (G(0)/G(1)) to DNA synthesis (S) and mitotic phase (G(2)/M) was held back by iptakalim in a concentration-dependent manner. Glyburide abolished the inhibitory effect of iptakalim. In conclusion, we have shown that iptakalim had an inhibitory effect on [Ca(2+)](cyt) increase and the proliferation of pulmonary arterial SMCs induced by endothelin-1 through activation of K(ATP) channels. These findings suggest that iptakalim might be a promising candidate for the treatment of pulmonary arterial remodeling in pulmonary hypertension. PMID- 15792776 TI - Induction of human leukemia HL-60 cell differentiation via a PKC/ERK pathway by helenalin, a pseudoguainolide sesquiterpene lactone. AB - Helenalin, a cell-permeable pseudoguainolide sesquiterpene lactone, is a potent anti-inflammatory agent that inhibits nuclear factor-kappa B (NF-kappa B) DNA binding activity. In this report, we investigated the effect of helenalin on cellular differentiation in the human promyelocytic leukemia HL-60 cell culture system. Helenalin by itself markedly induced HL-60 cell differentiation in a concentration-dependent manner. Cytofluorometric analysis and cell morphologic studies indicated that helenalin induced cell differentiation predominantly into granulocytes. Protein kinase C (PKC) and extracellular signal-regulated kinase (ERK) inhibitors significantly inhibited HL-60 cell differentiation induced by helenalin, while p38 mitogen-activated protein kinase (MAPK) inhibitors did not. Moreover, helenalin enhanced PKC activity and protein level of PKC beta I and PKC beta II isoforms, and also increased the level of pERK in a concentration dependent manner. In addition, the enhanced levels of cell differentiation closely correlated with the decreased levels of NF-kappa B binding activity by helenalin. These results indicate that PKC, ERK, and NF-kappa B may be involved in HL-60 cell differentiation induced by helenalin. PMID- 15792777 TI - Effects of seven drugs of abuse on action potential repolarisation in sheep cardiac Purkinje fibres. AB - Seven drugs of abuse have been examined for effects on the action potential in sheep isolated cardiac Purkinje fibres. Phencyclidine (5 microM) induced a significant increase (30.7%) in action potential duration at 90% repolarisation (APD(90)). Similarly, 10 microM 3,4-methylenedioxymethamphetamine (MDMA, 'Ecstasy') induced a significant increase in APD(90) of 12.1%. Although Delta(9) tetrahydrocannabinol (0.1 microM) induced a small, but statistically significant, 4.8% increase in APD(90), no effects were observed at 0.01 or 1 microM. Cocaethylene (10 microM) induced a significant shortening of APD(90) (-23.8%). Cocaine (up to 1 microM), (+)-methamphetamine ('Speed'; up to 5 microM), and the heroin metabolite, morphine (up to 5 microM), had no statistically significant effects. The possible significance of these findings is discussed in the context of other recognised cardiac effects of the tested drugs. PMID- 15792778 TI - Direct, pleiotropic protective effect of cyclosporin A against simulated ischemia induced injury in isolated cardiomyocytes. AB - Cyclosporin A is an immunosuppressor that prolongs graft survival but its use is limited by cardiotoxicity. The effects of cyclosporin A on several functional and biological characteristics were thus evaluated in rat cardiomyocytes in normal conditions and in a substrate-free, hypoxia-reoxygenation model of ischemia reperfusion. Cyclosporin A (100 and 1000 ng/ml) did not induce cardiocytotoxicity in basal conditions. Simulated ischemia gradually decreased and then blocked the spontaneous electromechanical activity. Cyclosporin A at 100 and 1000 ng/ml permitted the maintenance of electromechanical functions that were abolished in control cells. Cyclosporin A also improved the post-"ischemic" functional recovery. Cyclosporin A reduced the "ischemia"-induced lactate dehydrogenase and troponine I releases and the successive rises in heat shock protein mRNA observed after "ischemia" and reoxygenation. Moreover, cyclosporin A improved the resumption of the mitochondrial function. To conclude, cyclosporin A displayed a direct, pleiotropic protection of isolated cardiomyocytes against physiological, metabolic, structural and stress signaling changes induced by ischemia reperfusion mimicked in vitro. PMID- 15792779 TI - Estrogen mediated inhibition of dopamine transport in the striatum: regulation by G alpha i/o. AB - In the current study, the interaction between estrogen priming and dopamine D2 receptor activation on dopamine uptake in the striatum of ovariectomized female rats was investigated. Basal ADP-[(32)P(i)]ribosylation of G(i/o) was examined in synaptosomal membranes prepared from ovariectomized, estrogen primed or N-p (isothiocyanatophenethyl) spiperone (NIPS) treated rats. [(32)P(i)]-incorporation was significantly increased (141%) in tissue from NIPS treated animals but attenuated (57%) in tissue from estrogen primed animals. Dopamine uptake kinetics were measured in vivo following manipulation of the heterotrimeric G-protein by pertussis toxin (0.5 microg, 48 h). Pertussis toxin significantly inhibited dopamine uptake at all concentrations of dopamine examined. Co-treatment with estrogen and pertussis toxin resulted in a further attenuation of dopamine transport at high but not low dopamine concentrations. These data are consistent with an estrogen mediated alteration of G-protein activity and support the hypothesis that estrogen may alter transporter activity through a modulation of dopamine D2 autoreceptor/G alpha(i/o) protein coupling. PMID- 15792780 TI - In vivo neuroprotective effects of the novel imidazolyl nitrone free-radical scavenger (Z)-alpha-[2-thiazol-2-yl)imidazol-4-yl]-N-tert-butylnitrone (S34176). AB - Herein, we report an extensive investigation of the neuroprotective effects of the compound (Z)-alpha-[2-thiazol-2-yl)imidazol-4-yl]-N-tert-butylnitrone (S34176) and the prototypic nitrone alpha-phenyl-N-tert-butylnitrone (PBN), in different in vivo paradigms of neuronal degeneration. Administration of S34176 (75 mg/kg i.p.) 30 min before transient (10 min) global ischaemia in Wistar rats significantly prevented delayed neuronal cell death in the hippocampal CA1 area 7 days post-ischaemia (24% vs. 73% in ischaemia control; P<0.05) whereas PBN was inactive under similar conditions. Furthermore, oral administration of S34176 (30 mg/kg) 60 min before and during (1 x 30 mg/kg p.o.) 6 days post-ischaemia, in combination with an acute post-ischaemia sub-protective dose (3 x 10 mg/kg i.p.) of the glutamate receptor antagonist, 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo benzo[f]quinoxaline-7-sulfonamide (NBQX), resulted in an increased neuroprotective action (29% cell loss in drug-treated vs. 84% in ischaemia control P<0.001) compared to either compound alone. S34176 (20 mg/kg i.p.) also partially prevented kainic acid-induced neuronal cell death at 7 days post exposure in the CA1 (41% in drug-treated vs. 74% for kainate-treated controls; P<0.01) and CA3 hippocampal region (22% vs. 53%; P<0.01). Under similar conditions, S34176 administered orally (40 mg/kg) produced a more marked protection against kainate-induced neuronal cell loss in the CA1 (13% in drug treated vs. 82%; P<0.001) and CA3 areas (10% vs. 52%; P<0.001). Sub-chronic oral administration of S34176 (10 mg/kg) also partially reduced kainate-induced hippocampal cell death in the CA1 (53% vs. 77%; P<0.01) and CA3 (23% vs. 53%; P<0.01) areas. Dopamine depletion in the striatum of C57BL/6 mice induced by systemic D-methamphetamine injection was significantly reduced by S34176 (40+/-5% vs. 11.5+/-8%; P<0.001) (150 mg/kg i.p.) whereas PBN was inactive under similar conditions. S34176 represents a new centrally acting nitrone-based radical scavenger with neuroprotective properties in in vivo models of delayed neuronal cell death, and supports the therapeutic potential of this class of compound for the treatment of cerebral pathologies implicating chronic neurodegeneration. PMID- 15792781 TI - Interaction between inhibitors of inducible nitric oxide synthase and cyclooxygenase in Brewer's yeast induced pyrexia in mice: an isobolographic study. AB - We studied the interaction of S-methylisothiourea (a selective inducible nitric oxide synthase inhibitor) with rofecoxib (selective cyclooxygenase-2 inhibitor) and mefenamic acid (non-selective cyclooxygenase inhibitor) in Brewer's yeast induced pyrexia in mice by isobolographic analysis. Each drug was effective in reducing pyrexia when used alone. Log-dose-response curves of all the three drugs did not show any significant departure from parallelism indicating thereby, a common mode of antipyretic action. However, rofecoxib exhibited significantly higher potency than S-methylisothiourea. Isobolographic analysis of combination of S-methylisothiourea with rofecoxib and mefenamic acid revealed additive interaction. Experimental ED(50) of the combinations was not significantly different from theoretical additive ED(50) of the corresponding drug combination, that substantiated the additive nature of interaction between inducible nitric oxide synthase and cyclooxygenase in Brewer's yeast-induced fever in mice. Results suggest involvement of a mediator that is subservient to both inducible nitric oxide synthase and cyclooxygenase-2 enzyme activities. For further investigation, peroxynitrite ion may be considered to be the putative mediator. PMID- 15792782 TI - Nicotine and epibatidine alter differently nomifensine-elevated dopamine output in the rat dorsal and ventral striatum. AB - We studied the effects of nicotine and epibatidine given in combination with dopamine uptake inhibitor, nomifensine, on striatal extracellular dopamine and its metabolites by using brain microdialysis in freely moving rats. Nomifensine (3 mg/kg) elevated extracellular dopamine in the caudate-putamen, and clearly more in the nucleus accumbens. In the caudate-putamen, nicotine (0.5 mg/kg) and epibatidine (0.6 microg/kg but not 3.0 microg/kg) enhanced nomifensine's effect on dopamine. The effect of nomifensine on accumbal dopamine was enhanced by nicotine, but inhibited by epibatidine at 0.6 microg/kg. The larger dose of epibatidine had no effect. Thus, the effects of the smaller epibatidine dose (0.6 microg/kg) on the dopamine output in the caudate-putamen but not in the accumbens resemble those of nicotine 0.5 mg/kg. Discrepancies in the effects of epibatidine and nicotine are most probably due to differences in their affinities to nicotinic receptor subtypes regulating dopamine release. Further, different responses to low concentrations of epibatidine between the brain areas suggest that there are differences in the nicotinic regulation of nigrostriatal and mesolimbic dopaminergic pathways. PMID- 15792783 TI - Memory facilitation by methylene blue: dose-dependent effect on behavior and brain oxygen consumption. AB - Methylene blue administered post-training improves memory retention in avoidance and appetitive tasks, and restores spatial memory impaired by an inhibitor of cytochrome oxidase. Methylene blue may improve memory retention by increasing brain oxygen utilization. We investigated which doses improve memory without nonspecific behavioral effects, and whether methylene blue enhances brain oxygen consumption. Different doses were evaluated 24 h after administration in wheel running, feeding, open field habituation and object recognition tests. The 1-10 mg/kg methylene blue-treated rats were not different from saline-treated rats in locomotion or feeding behavior. The 50-100 mg/kg doses decreased running wheel behavior. The 4 mg/kg dose improved behavioral habituation and object memory recognition. Dose-dependent effects of methylene blue on brain oxygen consumption revealed that low concentrations increased brain oxygen consumption in vitro and 24 h after in vivo administration. Therefore, methylene blue doses that increase brain oxygen consumption also facilitate memory retention. PMID- 15792784 TI - Post-training intrahippocampal infusion of the COX-2 inhibitor celecoxib impaired spatial memory retention in rats. AB - In this study, we investigated the effects of intrahippocampal infusion of indomethacin as a non-selective cyclooxygenase inhibitor and celecoxib as a selective cyclooxygenase-2 inhibitor on spatial memory in the Morris water maze. Rats were trained for 3 days; each day included two blocks, and each block contained 4 trials. Tests were performed 48 h after surgery. Bilateral intrahippocampal infusion of indomethacin (0.01, 0.1, or 1 M) did not show any significant effect on spatial memory retention at these concentrations in rats. We also examined effects of infusion of celecoxib (0.02, 0.06, or 0.1 M) on memory retention. Bilateral infusion of 0.1 M celecoxib significantly altered escape latency and traveled distance in rats. These results strongly suggest that cyclooxygenase-2 is involved in spatial memory retention. PMID- 15792785 TI - Involvement of nitric oxide in both central and peripheral haemodynamic effect of D/L-nebivolol and its enantiomers in rats. AB - The cardiovascular profile of the racemate D/L-nebivolol and its enantiomers administered by intravenous (i.v.) or by intracerebroventricular (i.c.v.) route was investigated in anaesthetized normotensive rats. D/L-Nebivolol (0.1-0.5 mg/kg) induced a dose-related reduction in blood pressure when administered by i.c.v. route. These hypotensive effects were more marked as compared to those achieved by peripheral administration of D/L-nebivolol (0.1-1 mg/kg i.v.). Both enantiomers contributed to the hypotensive effect of D/L-nebivolol by i.c.v. route, while the effects of the drug on blood pressure by i.v. route were due to the d-enantiomer. The bradycardic effect of the racemic form given i.v. was dose related and, at the highest dose (1 mg/kg), was more pronounced as compared to i.c.v. route. D-Nebivolol was responsible for chronotropic effects by both the i.v. and i.c.v. route, although by i.c.v. route L-nebivolol also induced a reduction in heart rate. The nitric oxide synthase inhibitor N(omega)-nitro-L arginine methyl ester (L-NAME) administered at 5 mg/kg i.v. bolus + 0.1 mg/kg/min infusion or at 2.5 mg/kg i.c.v. counteracted the effects of D/L-nebivolol (either 1 mg/kg i.v. or 0.5 mg/kg i.c.v.) on blood pressure, while it did not inhibit the cardiovascular changes induced by isoprenaline (300 ng/kg i.v.) or calcitonin gene-related peptide (CGRP; 400 ng/kg i.v.). In addition, i.c.v. effects of D/L nebivolol on blood pressure and heart rate were not affected by pre-treatment with atropine (2 mg/kg i.v.). The present findings demonstrate that D/L-nebivolol produced haemodynamic changes following both peripheral and central administration; these latter findings are mainly due to its L-enantiomer and these effects involve the L-arginine/nitric oxide pathway. PMID- 15792786 TI - Effects of KR-32570, a new Na+/H+ exchanger inhibitor, on functional and metabolic impairments produced by global ischemia and reperfusion in the perfused rat heart. AB - The present study was performed to evaluate the cardioprotective effects of [5-(2 methoxy-5-chloro-5-phenyl)furan-2-ylcarbonyl]guanidine (KR-32570) on ischemia/reperfusion-induced mechanical and metabolic dysfunction in isolated rat hearts. In addition, the effects of KR-32570 on the Na(+)/H(+)-exchanger (NHE) and lipid peroxidation were also evaluated. KR-32570 strongly inhibited the recovery from acidosis induced by an NH(4)Cl prepulse in PS120 fibroblast cells expressing the human NHE-1 isoform (IC(50): 0.05 and 1.16 microM for KR-32570 and cariporide, respectively). In isolated perfused rat hearts subjected to 30-min ischemia/30-min reperfusion, KR-32570 (1-10 microM) significantly and concentration dependently improved cardiac contractile function and severe contracture in conjunction with causing a marked reduction in lactate dehydrogenase release. Additionally, it (1-10 microM) significantly increased the content of ATP, creatine phosphate and glycogen as well as decreased the tissue lactate content in heart homogenates following ischemia and reperfusion. KR-32570 (1-10 microM) significantly decreased the concentration of 8-iso-prostaglandin F(2 alpha), a reliable marker for oxidant stress, in perfusates from rat hearts subjected to ischemia and reperfusion. In separate experiments, KR-32570 significantly lowered the concentration of malondialdehyde in rat liver homogenate and inhibited Cu(2+)-induced peroxidation of low-density lipoprotein. Taken together, these results suggest that KR-32570 possesses potent cardioprotective effects in perfused rat hearts, and its effects may be mediated by inhibition of NHE-1, preservation of high-energy phosphates, and inhibition of lipid peroxidation. PMID- 15792787 TI - Morphine modulates inducible nitric oxide synthase expression and reduces pulmonary oedema induced by alpha-naphthylthiourea. AB - This study was designed to investigate the possible participation of morphine in pulmonary oedema induced by alpha-naphthylthiourea (ANTU), which is a well-known noxious chemical agent in the lung. Injection of ANTU (15 mg/kg i.p.) produced pulmonary oedema as indicated by an increase in lung weight/body weight ratio and pleural effusion reaching a maximum within 4 h in rat. Administration of morphine prior to ANTU significantly inhibited to pulmonary oedema with a dose-dependent manner. The protective effect of morphine is prevented by peripheral opioid receptor antagonist, naloxone methiodide. ANTU-treated rats were shown positive by inducible nitric oxide synthase immunohistochemical staining. There was no staining in the control group. On the other hand, the degree of staining was markedly reduced in tissue sections by morphine. These results suggest that previous administration of subcutaneous morphine has preventive effect on ANTU induced pulmonary inflammatory reaction and its effect mediated via peripheral opioid receptors. Application of naloxone with ANTU has no effect on the lung parameters indicating that endogenous opioids do not modulate ANTU-induced damage. PMID- 15792788 TI - Subconjunctivally administered celecoxib-PLGA microparticles sustain retinal drug levels and alleviate diabetes-induced oxidative stress in a rat model. AB - We have previously reported that repeated oral doses of celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, reduced diabetes-induced retinal vascular endothelial growth factor (VEGF) expression [Ayalasomayajula, S.P., Kompella, U.B., 2003. Celecoxib, a selective cyclooxygenase-2 inhibitor, inhibits retinal vascular endothelial growth factor expression and vascular leakage in a streptozotocin-induced diabetic rat model. Eur J Pharmacol 458, 283-289] and that retinal celecoxib delivery can be improved by several-fold following subconjunctival administration [Ayalasomayajula, S.P., Kompella, U.B., 2004. Retinal delivery of celecoxib is several-fold higher following subconjunctival administration compared to systemic administration. Pharm Res 21, 1797-1804]. The objective of the current study was to determine whether polymeric microparticles of celecoxib sustain retinal drug levels following subconjunctival administration and alleviate diabetes-induced oxidative stress in a streptozotocin-induced diabetic rat model. Biodegradable poly (lactide-co-glycolide) (PLGA; 85:15) microparticles of celecoxib were prepared using solvent evaporation method and characterized for their size, morphology, encapsulation efficiencies, and in vitro release. The celecoxib-PLGA microparticles or solution containing 75 microg of celecoxib was administered subconjunctivally to one eye (ipsilateral) of Sprague Dawley rats and drug levels in the retina, vitreous, lens, and cornea of ipsilateral and contralateral eyes were determined on 1, 7, and 14 days using high-performance liquid chromatography (HPLC). The effect of subconjunctivally administered celecoxib-PLGA microparticles on oxidative stress in day 14 diabetic rat retinas was determined by measuring the retinal glutathione (reduced (GSH) and oxidized (GSSG)), thiobarbituric acid reactive substances, and 4 hydroxynonenal levels using spectrofluorometric and colorimetric methods. Solvent evaporation method produced spherical celecoxib-PLGA microparticles with mean diameters of 3.9+/-0.6 microm and 68.5% loading efficiency. These microparticles sustained celecoxib release during the 49-day in vitro release study. Subconjunctivally administered celecoxib-PLGA microparticles sustained retinal and other ocular tissue drug levels during the 14-day study in rats. No detectable celecoxib levels were observed in the contralateral eye. The celecoxib PLGA microparticles significantly inhibited the diabetes-induced increases in thiobarbituric acid reactive substance (P=0.012) and 4-hydroxynonenal levels (P=0.029). The particles also inhibited the GSH depletion and the increase in GSSH/GSH ratio associated with diabetes but the effects were not statistically significant (P=0.12). Thus, following subconjunctival administration, celecoxib PLGA microparticles sustained retinal celecoxib delivery and inhibited diabetes induced retinal oxidative damage, indicating their potential usefulness in treating diabetes-induced retinal abnormalities. PMID- 15792789 TI - Differential respiratory effects of [Dmt1]DALDA and morphine in mice. AB - H-Dmt-D-Arg-Phe-Lys-NH(2) ([Dmt(1)]DALDA, dDAL), a highly selective mu-opioid peptide, produces potent analgesia without respiratory depression after intrathecal administration. Despite carrying 3+ net charge, dDAL is also a potent analgesic after systemic administration. We compared the respiratory effects of dDAL and morphine after subcutaneous administration in mice using whole body plethysmography. Analgesic doses of 3 and 10 times ED(50) were examined. Both drugs dose-dependently decreased respiratory frequency and minute volume in room air. Tidal volume was increased by the lower dose of morphine, while it was decreased by the higher dose of dDAL. The decrease in minute volume by dDAL and morphine was completely reversed by naloxone. No difference in ventilatory response to CO(2) was observed between dDAL and morphine at three times ED(50). Ventilatory response to hypoxia was significantly diminished by dDAL compared to morphine and saline, and this effect of dDAL was naloxone-irreversible. Thus dDAL likely reduces the sensitivity of the peripheral chemoreflex loop through a non opioid action. PMID- 15792790 TI - Anti-exudative effects of opioid receptor agonists in a rat model of carrageenan induced acute inflammation of the paw. AB - We evaluated the anti-exudative effects (Evan's blue) of mu-, delta- and kappa opioid receptor agonists in a rat model of carrageenan-induced acute inflammation. The contribution of different components was assessed after the administration of: cyclosporine A, capsaicin, 6-hydroxydopamine, compound 48/80, and specific histamine-receptor antagonists. The results show that the mu-opioid receptor agonists morphine and fentanyl and the delta-opioid receptor agonists DPDPE (enkephalin, [D-Pen(2,5)]) and SNC 80 ((+)-4-[(alpha R)-alpha((2S,5R)-4 allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N diethylbenzamide) decrease plasma extravasation in a dose-dependent manner, with a biphasic response. The effects were reversed by specific antagonists, and are predominantly mediated by peripheral opioid receptors. The integrity of sensory and sympathetic fibres is essential for the anti-exudative effects of fentanyl and DPDPE. Histamine and functional histamine H(2) and H(3) receptors are required for morphine and fentanyl (but not DPDPE) inhibition of plasma extravasation, suggesting different mechanism for mu- and delta-opioid receptor agonists. The present findings implicate multiple sites and mechanisms in the anti-exudative effects of exogenous opioids. PMID- 15792793 TI - Are splicing mutations the most frequent cause of hereditary disease? AB - Disease-causing point mutations are assumed to act predominantly through subsequent individual changes in the amino acid sequence that impair the normal function of proteins. However, point mutations can have a more dramatic effect by altering the splicing pattern of the gene. Here, we describe an approach to estimate the overall importance of splicing mutations. This approach takes into account the complete set of genes known to be involved in disease and suggest that, contrary to current assumptions, many mutations causing disease may actually be affecting the splicing pattern of the genes. PMID- 15792791 TI - Evaluating the prophylactic potential of the phtalimide derivative LASSBio 552 on allergen-evoked inflammation in rats. AB - A previous study showed that the novel tetrazolephtalimide derivative LASSBio 552 (2-4-[3-(1H-1,2,3,4-tetraazol-5-yl)propoxy]phenethyl-1,3-isoindolinedione) prevents LTD(4)-evoked tracheal contraction. This led us to examine the putative anti-inflammatory effect of LASSBio 552 in comparison with the leukotriene CysLT(1) receptor antagonist zafirlukast using a model of allergic pleurisy in rats. Treatment with either LASSBio 552 (24-96 micromol/kg, i.p.) or zafirlukast (9-72 micromol/kg, i.p.), 1 h before challenge, inhibited eosinophil and mononuclear cell influx into the pleural cavity 24 h post-challenge, but failed to alter the increased levels of eotaxin, plasma leakage, mast cell degranulation and neutrophil infiltration noted 6 h post-challenge. CD4(+) T cell recruitment 24 h post-challenge was also sensitive to LASSBio 552. This treatment failed to alter cysteinyl leukotriene production at 6 h, but clearly inhibited the phenomenon 24 h and 48 h post-challenge. In in vitro settings LASSBio 552 inhibited allergen-evoked cysteinyl leukotriene generation from isolated mast cells, while histamine release remained unchanged. It also slightly inhibited cysteinyl leukotriene production by eosinophils and mononuclear cells triggered by Ca(+2) ionophore A23187. A leukotriene CysLT(1) receptor transfected cell based assay revealed that LASSBio 552 did not prevent LTD(4)-evoked Ca(+2) influx, indicating that it was not a leukotriene CysLT(1) receptor antagonist. These findings indicate that LASSBio 552 is able to inhibit eosinophil influx triggered by allergen chalenge in a mechanism at least partially associated with suppression of CD4(+) T cell influx and cysteinyl leukotriene production. PMID- 15792794 TI - ERK-dependent induction of TNFalpha expression by the environmental contaminant benzo(a)pyrene in primary human macrophages. AB - Polycyclic aromatic hydrocarbons (PAHs) such as benzo(a)pyrene (BP) are toxic environmental contaminants known to enhance production of pro-inflammatory cytokines such as IL-1beta. The present study was designed in order to determine whether TNFalpha, another cytokine acting in inflammation, may also constitute a target for these chemicals. Both TNFalpha mRNA and TNFalpha secretion levels were found to be enhanced in human BP-treated macrophages. Dioxin, a contaminant activating the aryl hydrocarbon receptor (AhR) like PAHs, was also shown to increase TNFalpha expression. BP-mediated TNFalpha induction was however not suppressed by AhR antagonists, making unlikely the involvement of the typical AhR signalling pathway. BP-exposure of macrophages did not enhance NF-kappaB DNA binding activity, but it activated the MAP kinase ERK1/2. In addition, the use of chemical inhibitors of extracellular signal-regulated protein kinase (ERK) activation fully abrogated induction of TNFalpha production in BP-treated macrophages. These data likely indicate that PAHs enhance TNFalpha expression in human macrophages through an ERK-related mechanism. Such a regulation may contribute to confer pro-inflammatory properties to these widely-distributed environmental contaminants. PMID- 15792795 TI - Similarities and differences in the transcriptional regulation of the leptin gene promoter in gastric and adipose cells. AB - The stomach was reported to synthesize and secrete leptin mainly in the gastric lumen. Gastric leptin release is markedly increased after food intake, by vagal cholinergic stimulation and by cholecystokinin and secretin. Here we show that human gastric MKN-74 cells produce leptin that increases upon challenge with cholecystokinin, insulin, glucocorticoids and all-trans retinoic acid through activation of the leptin gene promoter. In addition, we demonstrate that forskolin and BRL37344 which increased cAMP levels, fail to affect the activity of leptin gene promoter in MKN74 expressing beta(3)-adrenoceptor cells but, induce a 2-fold decrease in this activity in adipose 3T3-L1 cells. These data described for the first time, similarities and more interestingly, differences in the regulation of the leptin gene promoter in gastric cells as compared to adipocytes. PMID- 15792796 TI - Isolation and structural characterization of epilancin 15X, a novel lantibiotic from a clinical strain of Staphylococcus epidermidis. AB - The potential application of lantibiotics as food-preserving agents and more recently as antibiotics has strongly increased the interest in these antibacterial peptides. Here, we report the elucidation of the primary and three dimensional structures of the novel lantibiotic epilancin 15X from Staphylococcus epidermidis using high-resolution nuclear magnetic resonance spectroscopy and tandem mass spectrometry. The molecule contains ten post-translationally modified amino acids, three lanthionine ring structures and a hydroxy-propionyl N-terminal moiety. The primary and tertiary structure and the distribution of positive charges are closely similar to the previously identified lantibiotic epilancin K7, most likely indicative of a common mode of action. PMID- 15792797 TI - Regulation of divalent metal transporter expression in human intestinal epithelial cells following exposure to non-haem iron. AB - A number of regulatory factors including dietary iron levels can dramatically alter the expression of the intestinal iron transporter DMT1. Here we show that Caco-2 cells exposed to iron for 4h exhibited a significant decrease in plasma membrane DMT1 protein, though total cellular DMT1 levels were unaltered. Following biotinylation of cell surface proteins, there was a significant increase in intracellular biotin-labelled DMT1 in iron-exposed cells. Furthermore, iron-treatment increased levels of DMT1 co-localised with LAMP1, suggesting that the initial response of intestinal epithelial cells to iron involves internalisation and targeting of DMT1 transporter protein towards a late endosomal/lysosomal compartment. PMID- 15792798 TI - Mitochondrial localization of Arabidopsis thaliana Isu Fe-S scaffold proteins. AB - Isu are scaffold proteins involved in iron-sulfur cluster biogenesis and playing a key role in yeast mitochondria and Escherichia coli. In this work, we have characterized the Arabidopsis thaliana Isu gene family. AtIsu1,2,3 genes encode polypeptides closely related to their bacterial and eukaryotic counterparts. AtIsu expression in a Saccharomyces cerevisiae Deltaisu1Deltanfu1 thermosensitive mutant led to the growth restoration of this strain at 37 degrees C. Using Isu GFP fusions expressed in leaf protoplasts and immunodetection in organelle extracts, we have shown that Arabidopsis Isu proteins are located only into mitochondria, supporting the existence of an Isu-independent Fe-S assembly machinery in plant plastids. PMID- 15792799 TI - All three J-domain proteins of the Escherichia coli DnaK chaperone machinery are DNA binding proteins. AB - DnaJ, DjlA and CbpA are the J-domain proteins of DnaK, the major Hsp70 of Escherichia coli. CbpA was originally discovered as a DNA binding protein. Here, we show that DNA binding is a property of DnaJ and DjlA as well. Of special interest in this respect is DjlA, as this cytoplasmic protein is membrane bound and, as shown here, its affinity for DNA is extremely high. The finding that all the three J-proteins of DnaK are DNA binding proteins sheds new light on the cellular activity of these proteins. PMID- 15792800 TI - Selection for TRAIL resistance results in melanoma cells with high proliferative potential. AB - To better understand the outcome of the interaction between TNF-related apoptosis inducing factor (TRAIL) and tumor cells, we studied TRAIL-resistant melanoma cells resulting from prolonged exposure to TRAIL and found that they had higher proliferative activity than the parental cells both in vitro and in vivo. This was associated with reduced p53 and p21 expression and increased activation of Erk1/2 and Akt. Accelerated proliferation was not due to TRAIL-mediated signaling but appeared to be the result of selection of previously existing, characteristically distinct cells. Moreover, responses of p53 to stimulation in the TRAIL-resistant cells appeared to be impaired. PMID- 15792801 TI - Hepsin activates pro-hepatocyte growth factor and is inhibited by hepatocyte growth factor activator inhibitor-1B (HAI-1B) and HAI-2. AB - Hepsin, a type II transmembrane serine protease, is highly upregulated in prostate cancer and promotes tumor progression and metastasis. We generated a soluble form of hepsin comprising the entire extracellular domain to show that it efficiently converts single-chain hepatocyte growth factor (pro-HGF) into biologically active two-chain HGF. Hepsin activity was potently inhibited by soluble forms of the bi-Kunitz domain inhibitors HAI-1B (IC(50) 21.1+/-2.7 nM) and HAI-2 (IC(50) 1.3+/-0.3 nM). Enzymatic assays with HAI-1B Kunitz domain mutants (R260A and K401A) further demonstrated that inhibition was due to Kunitz domain-1. The results suggest a functional link between hepsin and the HGF/Met pathway, which may contribute to tumor progression. PMID- 15792803 TI - Evidence for major structural changes in subunit C of the vacuolar ATPase due to nucleotide binding. AB - The ability of subunit C of eukaryotic V-ATPases to bind ADP and ATP is demonstrated by photoaffinity labeling and fluorescence correlation spectroscopy (FCS). Quantitation of the photoaffinity and the FCS data indicate that the ATP analogues bind more weakly to subunit C than the ADP-analogues. Site-directed mutagenesis and N-terminal sequencing of subunit C from Arabidopsis (VHA-C) and yeast (Vma5p) have been used to map the C-terminal region of subunit C as the nucleotide-binding site. Tryptophan fluorescence quenching and decreased susceptibility to tryptic digestion of subunit C after binding of different nucleotides provides evidence for structural changes in this subunit caused by nucleotide-binding. PMID- 15792802 TI - Extracellular HSP70 binding to surface receptors present on antigen presenting cells and endothelial/epithelial cells. AB - Extracellular HSP70 has been found to participate in both innate and adaptive immune responses. However, little is known about the molecular mechanisms that mediate this process. Previous reports suggest that HSP70 interacts with antigen presenting cells (APC) through a plethora of surface receptors. In this study, we have examined the relative binding of potential HSP70 receptors and found high affinity binding to LOX-1 but not other structures with a role in HSP70-APC interactions such as LRP/CD91, CD40, TLR2, TLR4 or another c-type lectin family member (DC-SIGN) closely related to LOX-1. In addition to APC, HSP70 can avidly bind to non-APC cell lines, especially those from epithelial or endothelial background. PMID- 15792804 TI - Structural basis of gating of CNG channels. AB - Cyclic nucleotide-gated (CNG) ion channels, underlying sensory transduction in vertebrate photoreceptors and olfactory sensory neurons, require cyclic nucleotides to open. Here, we present structural models of the tetrameric CNG channel pore from bovine rod in both open and closed states, as obtained by combining homology modeling-based techniques, experimentally derived spatial constraints and structural patterns present in the PDB database. Gating is initiated by an anticlockwise rotation of the N-terminal region of the C-linker, which is then, transmitted through the S6 transmembrane helices to the P-helix, and in turn from this to the pore lumen, which opens up from 2 to 5A thus allowing for ion permeation. The approach, here presented, is expected to provide a general methodology for model ion channels and their gating when structural templates are available and an extensive electrophysiological analysis has been performed. PMID- 15792805 TI - SKN1, a novel plant defensin-sensitivity gene in Saccharomyces cerevisiae, is implicated in sphingolipid biosynthesis. AB - The antifungal plant defensin DmAMP1 interacts with the fungal sphingolipid mannosyl diinositolphosphoryl ceramide (M(IP)(2)C) and induces fungal growth inhibition. We have identified SKN1, besides the M(IP)(2)C-biosynthesis gene IPT1, as a novel DmAMP1-sensitivity gene in Saccharomyces cerevisiae. SKN1 was previously shown to be a KRE6 homologue, which is involved in beta-1,6-glucan biosynthesis. We demonstrate that a Deltaskn1 mutant lacks M(IP)(2)C. Interestingly, overexpression of either IPT1 or SKN1 complemented the skn1 mutation, conferred sensitivity to DmAMP1, and resulted in M(IP)(2)C levels comparable to the wild type. These results show that SKN1, together with IPT1, is involved in sphingolipid biosynthesis in S. cerevisiae. PMID- 15792806 TI - Cold exposure differently influences mitochondrial energy efficiency in rat liver and skeletal muscle. AB - This study deals with mitochondrial energy efficiency in liver and skeletal muscle mitochondria in 15 days cold exposed rats. Cold exposure strongly increases the sensitivity to uncoupling by added palmitate of skeletal muscle but not liver mitochondria, while mitochondrial energy coupling in the absence of fatty acids is only slightly affected by cold in liver and skeletal muscle. In addition, uncoupling protein 3 content does not follow changes in skeletal muscle mitochondrial coupling. It is therefore concluded that skeletal muscle could play a direct thermogenic role based on fatty acid-induced mild uncoupling of mitochondrial oxidative phosphorylation. PMID- 15792807 TI - In vitro activity of C-20 methyltransferase, BchU, involved in bacteriochlorophyll c biosynthetic pathway in green sulfur bacteria. AB - The activity of a methyltransferase, BchU, which catalyzes methylation at the C 20 position of chlorin ring in the biosynthetic pathway of bacteriochlorophyll c, was investigated in vitro. The bchU gene derived from the photosynthetic green sulfur bacterium, Chlorobium tepidum, was overexpressed in Escherichia coli as a His-tagged protein (His(6)-BchU), and the enzyme was purified. In the presence of S-adenosylmethionine, His(6)-BchU methylated zinc bacteriopheophorbide d at the C 20 position to give zinc bacteriopheophorbide c. Metal-free bacteriopheophorbide d could not be methylated by the BchU, indicating that the central metal in the chlorin should be required for the recognition by the BchU. PMID- 15792808 TI - Multiple-mutation at a potential ligand-binding region decreased allergenicity of a mite allergen Der f 2 without disrupting global structure. AB - We assessed the effect of multiple-mutations within one IgE-binding area on allergenicity of Der f 2. The triple-mutant of Der f 2, P34/95/99A, exhibited the most significant reduction of allergenicity and circular dichroism analysis showed that the global structure of Der f 2 was maintained in P34/95/99A. These results indicate that such a strategy is effective when designing allergen vaccines, which achieve less allergenicity for a broad population of patients without disrupting the global structure. Structurally, Der f 2 is a member of the MD-2 related lipid-recognition proteins. The sites for the triple-mutation located on the characteristically charged entrance of a cavity and corresponded to the regions critical to ligand-binding in the Niemann-Pick type 2 disease protein and MD-2. PMID- 15792810 TI - Actin microridges characterized by laser scanning confocal and atomic force microscopy. AB - We have characterized the cell surface of zebrafish stratified epithelium using a combined approach of light and atomic force microscopy under conditions which simulate wound healing. Microridges rise on average 100 nm above the surface of living epithelial cells, which correlate to bundles of cytochalasin B-insensitive actin filaments. Time-lapse microscopy revealed the bundles to form a highly dynamic network on the cell surface, in which bundles and junctions were severed and annealed on a time scale of minutes. Atomic force microscopy topographs further indicated that actin bundle junctions identified were of two types: overlaps and integrated end to side T- and Y-junctions. The surface bundle network is found only on the topmost cell layer of the explant, and never on individual locomoting cells. Possible functions of these actin bundles include cell compartmentalization of the cell surface, resistance to mechanical stress, and F-actin storage. PMID- 15792809 TI - Amyloid-beta peptides induce cell proliferation and macrophage colony-stimulating factor expression via the PI3-kinase/Akt pathway in cultured Ra2 microglial cells. AB - Alzheimer's disease is characterized by numerous amyloid-beta peptide (Abeta) plaques surrounded by microglia. Here we report that Abeta induces the proliferation of the mouse microglial cell line Ra2 by increasing the expression of macrophage colony-stimulating factor (M-CSF). We examined signal cascades for Abeta-induced M-CSF mRNA expression. The induction of M-CSF was blocked by a phosphatidylinositol 3 kinase (PI3-kinase) inhibitor (LY294002), a Src family tyrosine kinase inhibitor (PP1) and an Akt inhibitor. Electrophoretic mobility shift assays showed that Abeta enhanced NF-kappaB binding activity to the NF kappaB site of the mouse M-CSF promoter, which was blocked by LY294002. These results indicate that Abeta induces M-CSF mRNA expression via the PI3 kinase/Akt/NF-kappaB pathway. PMID- 15792811 TI - Serine utilization in mouse liver: influence of caloric restriction and aging. AB - The influence of caloric restriction (CR) on the activities of hepatic serine metabolizing enzymes in young (3 months) and old (30 months) mice was studied. Serine dehydratase (SDH) activity increased markedly with age in both diet groups and in old mice was higher in the CR group. No effects of CR were observed in the young. Serine:pyruvate transaminase (SPT) and glycerate kinase activities were unaffected by age and diet. However, glycerate dehydrogenase activity was decreased in old CR mice but not in young CR. The results of this study show that long-term CR influenced serine utilization only in the pathway catalyzed by SDH. This suggests that in mouse liver this pathway is critical for serine utilization in gluconeogenesis, while the SPT pathway plays a minor role. The increase in SDH activity with long-term CR is consistent with sustained increase in gluconeogenesis. PMID- 15792812 TI - Endopolyphosphatase in Saccharomyces cerevisiae undergoes post-translational activations to produce short-chain polyphosphates. AB - Endopolyphosphatase (Ppn), responsible for cleavage of long chain inorganic polyphosphate (poly P) of several hundred residues to generate progressively shorter chains, has been identified in mammalian cells and purified from Saccharomyces cerevisiae. Disruption of the encoding gene, PHM5, in S. cerevisiae resulted in a mutant that showed limited growth and failure to survive in a minimal medium. The limited digestion products of the yeast enzyme Ppn1 judged to be P(3) and P(60) have now, with the homogeneous enzyme and improved separation methods, been demonstrated to be P(i) and P(3). Ppn1, a homotetramer of a 35-kDa subunit, is of vacuolar origin and requires protease activation of a 78 kDa (674 aa) precursor polypeptide (prePpn1). The protease-processed Ppn1 has been purified 3800-fold to homogeneity and the protease cleavage sites determined. Both termini of prePpn1 and the post-translational modification of N glycosylations are essential for the protease-mediated maturation of Ppn1. PMID- 15792813 TI - Alzheimer's disease and vascular brain lesions. PMID- 15792814 TI - Alzheimer's disease with and without cerebral infarcts. AB - OBJECTIVE: To compare the clinical and pathological features of Alzheimer's disease (AD) patients with and without associated cerebral infarcts (CI). METHODS: The consecutive records of 57 prospectively studied demented patients fulfilling the CERAD criteria for the pathological diagnosis of AD were reviewed. Cases with cortical Lewy bodies were excluded. CI were found in 22 cases (39%) (AD+CI group): large infarcts (5), lacunes (13) and/or hippocampal sclerosis (4), and were absent in 35 cases (AD group). Microscopic infarcts, cribiform change, amyloid angiopathy, and white matter rarefaction were not considered in this classification, but were quantified. Cortical atrophy, neurofibrillary tangle and senile plaque (diffuse and neuritic) load were also measured. Pathological evaluation was independent of clinical information. Clinical and pathological data were compared between both groups. RESULTS: AD+CI cases were significantly older, more commonly female, less educated, and more often had blue collar occupations, sleep disturbances, frontal release signs, and EEG spikes than AD cases. Other differences found (acute/subacute onset, behavioral disturbances, and leukoaraiosis on CT scan) disappeared after controlling for age. The frequency of known vascular risk factors and focal motor and sensory signs did not differ between the groups, which showed remarkable clinical similarity overall. The only significant differences on pathological exam were hippocampal microinfarcts and white matter lesions, although there was a trend for lower neurodegenerative lesion load in the AD+CI group. The ischemic lesions were located in temporal lobe in 50% of AD+CI patients; these cases had a significantly lower neocortical neurodegenerative lesion load than those with CI in other sites. CONCLUSIONS: The presence of CI in AD increases significantly with age, but has scarce influence on the clinical features, and cannot be predicted from common vascular risk factors. In spite of a trend, there are no major differences in neurodegenerative lesion load between AD and AD+CI groups, except when CI are located in the temporal lobe (including hippocampus), suggesting that this location may be important in the physiopathology of mixed vascular and AD dementia. PMID- 15792815 TI - Amelioration by mecobalamin of subclinical carpal tunnel syndrome involving unaffected limbs in stroke patients. AB - Our previous study showed that overuse of the nonparetic hand and wrist of the nonparetic side following stroke result in significantly more abnormal on the nonparetic side than on the hemiparetic side in terms of electrophysiologic indices of median nerve function. The purpose of this study was to evaluate the effects of the orally administered mecobalamin, an analogue of vitamin B12, for carpal tunnel syndrome (CTS) in the nonparetic side in patients following stroke. In a randomized open label and prospective study of stroke patients, 67 received of 1500 mug mecobalamin daily for 2 years, and the remaining 68 (untreated group) did not. At baseline, sensory nerve conduction velocity, motor nerve conduction velocity, sensory nerve action potentials (SNAP) at the wrist, palm-to-wrist distal sensory latency, palm-to-wrist SNAP, motor nerve conduction velocity compound motor action potentials, and distal motor latency of median nerve were significantly more abnormal on the nonparetic side than on the hemiparetic side or in controls. Before the treatment 21 patients (31%) of untreated and 20 patients (30%) of treated group met electrophysiologic criteria for CTS. Sensory impairment of the nonparetic side had lessened in the treated group. After 2 years, all electrophysiologic indices of nonparetic side were significantly improved in the treated group compared with those in the untreated group. The improvement from baseline of electrophysiologic parameters in sensory nerve in the treated group was greater than the improvement measured in motor nerve. There were no side effects. Oral mecobalamin treatment is a safe and potentially beneficial therapy for CTS in stroke patients. PMID- 15792816 TI - Sensitivity and specificity of electrodiagnostic criteria for CIDP using ROC curves: comparison to patients with diabetic and MGUS associated neuropathies. AB - INTRODUCTION: Electrodiagnostic testing plays a key role in the characterization of neuropathies. To this end, sets of electrodiagnostic criteria have been proposed to define chronic inflammatory demyelinating polyneuropathy (CIDP). These criteria sets differ because of the number of data points within the sets, the number of required abnormal measures within a criterion, and the cutoff value of each measure. OBJECTIVE: To evaluate the sensitivity and specificity of the published criteria for defining CIDP in comparison to diabetic polyneuropathy (DMPN) and monoclonal gammopathy of undetermined significance associated neuropathies (MGUS-PN). DESIGN/METHODS: Electrodiagnostic studies of 21 patients with biopsy proven CIDP, 35 patients with MGUS-PN, and 82 patients with DMPN were analyzed. Data were compared against 4 different published criteria sets. Receiver operator characteristic (ROC) curves were used to determine the ideal threshold values for individual electrodiagnostic parameters. RESULTS/CONCLUSION: None of the currently published criteria sets could adequately separate CIDP from DMPN and MGUS-PN. Analysis of our data using ROC curves shows that the best discrimination was achieved using the following criteria: (1) FWL>145%ULN in one nerve or 110%ULN in two nerves in separate nerve roots. (2) Motor CV<90%LLN in four nerves with at least one nerve <70%LLN. Furthermore, proximal-to-distal amplitude ratio alone is not adequate to define conduction block. Although electrodiagnostic studies are important for evaluating CIDP, these studies by themselves cannot be used to define this neuropathy. PMID- 15792817 TI - Predicting quality of life in multiple sclerosis: accounting for physical disability, fatigue, cognition, mood disorder, personality, and behavior change. AB - Health-related quality of life (HQOL) is poor in multiple sclerosis (MS) but the clinical precipitants of the problem are not well understood. Previous correlative studies demonstrated relationships between various clinical parameters and diminished HQOL in MS. Unfortunately, these studies failed to account for multiple predictors in the same analysis. We endeavored to determine what clinical parameters account for most variance in predicting HQOL, and employability, while accounting for disease course, physical disability, fatigue, cognition, mood disorder, personality, and behavior disorder. In 120 MS patients, we measured HQOL (MS Quality of Life-54) and vocational status (employed vs. disabled) and then conducted detailed clinical testing. Data were analyzed by linear and logistic regression methods. MS patients reported lower HQOL (p<0.001) and were more likely to be disabled (45% of patients vs. 0 controls). Physical HQOL was predicted by fatigue, depression, and physical disability. Mental HQOL was associated with only depression and fatigue. In contrast, vocational status was predicted by three cognitive tests, conscientiousness, and disease duration (p<0.05). Thus, for the first time, we predicted HQOL in MS while accounting for measures from these many clinical domains. We conclude that self-report HQOL indices are most strongly predicted by measures of depression, whereas vocational status is predicted primarily by objective measures of cognitive function. The findings highlight core clinical problems that merit early identification and further research regarding the development of effective treatment. PMID- 15792818 TI - Comparison of age of onset and development of motor complications between smokers and non-smokers in Parkinson's disease. AB - BACKGROUND: There is growing evidence from case-control and from cohort studies that smoking is inversely related to the risk of developing Parkinson's disease (PD). However, it is still controversial if PD starts at an older age in ever smoking patients compared to never-smoking ones. PATIENTS AND METHODS: The present retrospective study compares in a large series of 512 out-patients, collected over the last 24 years, the age of onset of the complaints, the age at which PD was diagnosed and the start of levodopa treatment between ever- and never-smokers. Also, the occurrence of long-term side-effects of the drug was evaluated. 184 PD patients with a history of smoking were compared with 328 who had never smoked. The subgroups with and without a family history of PD were analysed separately. RESULTS: In the overall ever-smoking group, as well as in the subgroup without a family history, the onset of the disease and the time of the diagnosis of PD and the time at which levodopa was started occurred at an older age than in the never-smoking group. This difference could not be demonstrated in the patients with a family history, due to the low number of cases and the lack of statistical power. Although the follow-up period was the same in both study groups, motor fluctuations and dyskinesia were more frequent and appeared earlier after levodopa treatment in the non-smoking compared to the ever-smoking PD patients. Only for cognitive impairment there was a non significant trend in the smoking group. CONCLUSION: The present study confirms the protective action of smoking on PD and also suggests some modulating effect of smoking on the dopaminergic system. PMID- 15792819 TI - Serum uric acid levels and leukocyte nitric oxide production in multiple sclerosis patients outside relapses. AB - BACKGROUND: A number of studies found that patients with multiple sclerosis (MS) have low serum levels of uric acid. It is unclear whether this represents a primary deficit or secondary effect. Uric acid is a scavenger of peroxynitrite, which is the product of nitric oxide (NO) and superoxide. Because peripheral blood leukocyte NO production and NO metabolites in serum are raised in MS patients, associations might be expected between serum uric acid levels and peripheral NO production. METHODS: Serum levels of uric acid and NO production by peripheral blood leukocytes were measured in 60 patients with MS without a relapse in the past 3 months, and 30 age- and sex-matched healthy controls. Uric acid was determined with the uricase PAP method, and NO production was assayed by measuring nitrite concentration in supernatants of lysed leukocytes. RESULTS: Serum uric acid levels were not different between MS patients and controls. Compared to controls, patients with MS had significantly higher peripheral blood leukocytes nitrite concentrations (p<0.001). There was no correlation between leukocyte nitrite concentration and serum uric acid levels. CONCLUSIONS: Our findings suggest that in MS patients there is no primary deficit in serum uric acid. NO production by peripheral blood leukocytes is increased, but there is no association with serum uric acid levels. PMID- 15792820 TI - Videofluoroscopic assessment of swallowing function in patients with myasthenia gravis. AB - Swallowing function in myasthenia gravis (MG) was investigated by videofluoroscopy (VF). A total of 23 VF examinations were performed on 11 MG patients at various time points over the exacerbation and remission stages of disease. The assessment parameters on VF examination are set as follows: bolus transport from the mouth to the pharynx, bolus holding in the oral cavity, velopharyngeal seal, tongue base movement, pharyngeal constriction, laryngeal elevation, upper esophageal sphincter (UES) opening, and bolus stasis at the pyriform sinus (PS). Aspiration was also assessed on VF examination. Aspiration was seen in 34.8%, and half of these cases involved silent aspiration. Three of four cases that showed silent aspiration went on to experience aspiration pneumonia during the follow-up term. By Spearman's rank correlation, disturbance of laryngeal elevation was significantly correlated with aspiration (p=0.001), and incomplete UES opening was not significantly, but tended to be, correlated with aspiration (p=0.067). Although other parameters in the oral and pharyngeal phase on VF examination, such as bolus transport from the oral cavity to the pharynx, pharyngeal constriction, or stasis at the PS, were remarkably disturbed (in more than 50% of the examinations), those parameters were not good indicators for aspiration. When a disturbance of laryngeal elevation is found at a bedside clinical test, we recommend performing precise swallowing evaluation, such as VF, Fiberoptic Endoscopic Evaluation of Swallowing (FEES), and/or scintigraphic assessment of swallowing, for MG patients, to detect silent aspiration. PMID- 15792821 TI - Biochemical and behavioural phenotyping of a mouse model for GAMT deficiency. AB - Deficiency of guanidinoacetate N-methyltransferase (GAMT) is the first described creatine (CT) deficiency syndrome in man, biochemically characterized by accumulation of guanidinoacetic acid (GAA) and depletion of CT. Patients exhibit severe developmental and muscular problems. We created a mouse model for GAMT deficiency, which exerts biochemical changes comparable with those found in human GAMT-deficient subjects. CT and creatinine (CTN) levels are significantly decreased and GAA is increased in knockout (KO) mice. In patients, other guanidino compounds (GCs) appear to be altered as well, which may also contribute to the symptomatology. Extensive evaluation of GCs levels in the GAMT mouse model was therefore considered appropriate. Concentrations of 13 GCs in plasma, 24-h urine, brain and muscle of GAMT mice were measured. We also report on the detailed behavioural characterization of this model for GAMT deficiency. Besides an increase of GAA and a decrease of CT and CTN in plasma, 24-h urine, brain and muscle of KO mice, we observed a significant increase of other GCs in brain and muscle that was sometimes reflected in plasma and/or urine. KO mice displayed mild cognitive impairment. In general, it could be concluded that the GAMT mouse model is very useful for biochemical research of GAMT deficiency, but shows only a mild cognitive deficit. PMID- 15792822 TI - Cerebral PET imaging and histological evidence of transglutaminase inhibitor cystamine induced neuroprotection in transgenic R6/2 mouse model of Huntington's disease. AB - To investigate efficacy of cystamine induced neuroprotection, we conducted PET imaging studies of cerebral glucose metabolism with [(18)F]FDG (2-deoxy-2 [(18)F]fluoro-d-glucose) and striatal dopamine D2 receptor function with [(11)C]raclopride in R6/2 transgenic Huntington mice. In the control mice, exponentially decreasing glucose utilization was observed in the striatum N(str) [SUV]=(41.75+/-11.80)(58,str)*exp(-(0.041+/-0.007)*t [days]); cortex N(cort) [SUV]=24.14+/-3.66)(58,cort)*exp(-(0.043+/-0.007)*t [days]); and cerebellum N(cer) [SUV]=(34.97+/-10.58)(58,cer)*exp(-(0.037+/-0.008)*t [days]) as a function of age starting at 58 days. Given that the underlying degeneration rate in the cystamine treated mice is similar to that observed in control animals, the protection coefficient (beta) calculated from the equation N(t)=N(58)*exp(-(1 beta)*k*t) was 0.133+/-0.035 for the striatum; 0.122+/-0.028 for the cortex and 0.224+/-00.042 for the cerebellum with a dose of 100 mg/kg. The 50 mg/kg cystamine dose provided significant protection only for the striatum and only minor protection was obtained using lower doses. Striatal binding potential of [(11)C]raclopride was 1.059+/-0.030 in the control mice, and enhanced in the cystamine treated animals in a dose dependent manner up to 1.245+/-0.063 using the 100 mg/kg dose. Histological analysis confirmed cystamine induced neuroprotection of striatal and cortical neurons and Nissl staining revealed that formation of cellular inclusions was reversed in a dose dependent manner. Cerebral imaging and histological evidence support the use of cystamine as a neuroprotective agent for Huntington's disease (HD) pathology. PMID- 15792823 TI - Transcranial Doppler sonography evaluation of anterior cerebral artery hypoplasia or aplasia. AB - BACKGROUND AND PURPOSE: Transcranial Doppler (TCD) sonography is useful to evaluate intracranial arteries, however, interpretation of the TCD results in anterior cerebral artery (ACA) is difficult because of hypoplasia or aplasia. We try to define useful TCD indices and cut-off values to determine the variations of ACA. METHODS: Consecutive patients who underwent TCD and magnetic resonance angiography (MRA) were included. Patients with cerebrovascular abnormality or inadequate temporal windows were excluded. ACA status was classified as normal (NL), hypoplasia (HP), and aplasia (AP) according to MRA. TCD indices of mean flow velocity (MFV), pulsatility index (PI), ACA/middle cerebral artery (MCA) flow velocity ratio (ACA/MCA FVR), and asymmetry index (AI) of ACA were blindly compared with MRA between three groups. RESULTS: Two hundred and forty-one patients were included, and 193 patients (80%) were classified as NL, 34 (14%) as HP and 14 (6%) as AP. MFV was significantly lower in HP and AP (p<0.001), however, PI and ACA/MCA FVR were not different. AI was significantly different between NL and HP (21.5% vs. 50.4%), NL and AP (21.5% vs. 105.2%) (p<0.001). CONCLUSIONS: MFV of ACA should be interpreted with caution for its frequent anatomical variations. AI is useful to differentiate hypoplasia and aplasia from normal ACA with optimal criteria. PMID- 15792824 TI - Thiopurine methyltransferase (TPMT) heterozygosity and enzyme activity as predictive tests for the development of azathioprine-related adverse events. AB - Thiopurine methyltransferase (TPMT) is a key enzyme in azathioprine metabolism mediating both immunosuppression and cytotoxicity. TPMT activity may be influenced by a mutation in the TPMT gene resulting in individual differences in azathioprine metabolism. Individuals heterozygous for TPMT mutations or with low TPMT activity may be susceptible to azathioprine toxicity. We evaluate TPMT genotyping and TPMT enzyme activity as predictive tests for developing azathioprine-related adverse events. Neurological patients (n=129) observed whilst taking azathioprine therapy were genotyped for the TPMT*2, *3A and *3C mutations; TPMT enzyme activity was analysed in 92 of these patients. Ethnic appropriate controls (Black, Mixed-Ancestry and White) were genotyped (n=465) and of these controls TPMT activity was also measured in 115. Azathioprine-related adverse events developed in 21.7% of the patients; early (within 1 week) events included gastrointestinal symptoms (n=8/28). Haematological toxicity, hepatotoxicity, arthralgia, rashes and pancreatitis developing between 4 and 240 weeks. Genotyping showed that only four of 28 cases who developed adverse events, were heterozygous for TPMT*3A or *3C. Heterozygous patients developed either haematological or hepatic toxicity. In an ethnically heterogeneous society TPMT enzyme activity proved difficult to interpret as measurements amongst controls showed significant ethnic variation (p=1 x 10(-6)); cut-points between "low" and "normal" TPMT activity correlated with indigenous African genetic ancestry. Ethnic appropriate cut-points were determined but due to the ambiguity in interpreting TPMT enzyme results in heterogeneous societies, we favour genotyping as a predictive test. The positive predictive value of genotyping was low, but the likelihood ratio for developing either haematological or hepatotoxicity by identifying TPMT heterozygosity, was 9.75. In our patient population this translates into an improvement from a pre-test probability of developing haematological or hepatotoxicity of 11%, to a post-test level of 50%. Heterozygous patients may then be targeted for a more "tailored" increase in dosing and regular laboratory monitoring. PMID- 15792825 TI - Putaminal hemorrhage disrupts thalamocortical projection to secondary somatosensory cortex: case report. AB - Putaminal hemorrhage presenting pure sensory stroke is rare. We describe a case of left putaminal hemorrhage presenting contralateral hemisensory disturbance without hemiparesis. A 52-year-old man developed analgesia and thermoanesthesia in the right half of his body, but deep sensation was relatively well preserved. Neuroradiological and somatosensory evoked potential findings suggested that thalamocortical sensory pathways to the secondary somatosensory cortex (S2) were involved, whereas those to the primary somatosensory cortex (S1) were spared. In experimental animals, spinothalamic projections from the thalamic nucleus input directly to S2. In humans, thalamocortical pathways are still a subject of debate, but results of recent functional imaging studies suggest that the pathway of pain inputs directly to S2 and that of tactile sensation to S2 via S1. Our findings support these reports. PMID- 15792826 TI - Low-dose tacrolimus for two cases of myasthenia gravis with invasive thymoma that relapsed shortly after thymectomy. AB - Two patients with myasthenia gravis (Ossermann IIb) involving invasive thymoma who underwent extensive thymectomy manifested myasthenic crisis shortly after the procedure; however, both patients were treated with intravenous immunoglobulin and recovered from myasthenic crisis that had been deteriorating for about 1 week. Subsequently, the patients were administered a low-dose of tacrolimus (3 mg/day) in addition to prednisolone. Several months later, tacrolimus continued to control fluctuations of myasthenic symptoms and maintained remission in these patients. The serum titer of anti-Ach-receptor antibodies decreased in parallel with clinical improvement due to tacrolimus, and we accordingly reduced the dosage of prednisolone. Tacrolimus is a new immunosuppressive agent acting through the selective inhibition of helper-T-cell activation that can be reduced dosage of steroids and can maintain remission of myasthenia gravis with invasive thymoma. PMID- 15792827 TI - Chronic inflammatory demyelinating polyneuropathy in a patient with hyperIgEaemia. AB - We herein report the case of a 46 year old man with chronic inflammatory demyelinating polyneuropathy (CIDP) with hyperIgEaemia. The patient presented with bilateral weakness, generalized hyporeflexia, and mild paresthesia of the fingers of both hands. Nerve conduction studies revealed multiple sites of motor conduction block in the absence of sensory abnormalities. Muscle strength increased, as did compound muscle action potential (CMAP) amplitude immediately after the intravenous infusion of immunoglobulin (IVIg). Serum IgE levels also fluctuated in parallel with his relapsing-remitting clinical course. We propose that pure motor CIDP may be immune mediated and suggest that IgE-mediated allergy may be one potential cause of this condition. PMID- 15792828 TI - High-dose, frequently administered interferon beta. PMID- 15792830 TI - Re: Short-term brain atrophy changes in relapsing-remitting multiple sclerosis. PMID- 15792832 TI - Insulin and nitric oxide stimulates glucose transport in human placenta. AB - The present work examines whether insulin and NO can act as regulators of glucose transport in placenta. Glucose uptake (2-deoxy D-[(3)H]glucose) was measured in the absence (control or basal values) and in the presence of insulin (1200 microU/ml) or SNP (20 microM) in isolated perfused cotyledons and tissue slices preparations of human placenta. Both insulin and NO significantly increased glucose uptake by 20 and 27 per cent, respectively. Insulin decreased the Km of glucose transport from 42.5 +/- 2.69 to 35.1 +/- 2.58 mM. The stimulatory effect of SNP was mimicked by 8-CPT-cGMP and was completely blocked by the guanylate cyclase inhibitor, ODQ (10 microM). ODQ and the NOS inhibitor, L-NAME (100 microM), decreased basal glucose uptake but did not affect insulin-stimulated glucose transport. Taken together, these findings indicate that insulin and NO stimulate glucose uptake in human placenta and suggest that both potential regulators of glucose transport use different signaling pathways. PMID- 15792833 TI - Long-term treatment with the Na+-glucose cotransporter inhibitor T-1095 causes sustained improvement in hyperglycemia and prevents diabetic neuropathy in Goto Kakizaki Rats. AB - We examined the effects of T-1095, an orally active inhibitor of Na(+)-glucose cotransporter (SGLT), on the development and severity of diabetes in Goto Kakizaki (GK) rat, a spontaneous, non-obese model of type 2 diabetes. T-1095 was administered as dietary admixture (0.1% w/w) beginning at 7 weeks of age for 32 weeks. Untreated male GK rats were hyperglycemic compared with Wistar rats. Throughout the study, T-1095 treatment significantly decreased both blood glucose and hemoglobin A(1C) levels in the GK rats. The concomitant increase of urinary glucose excretion indicated that the hypoglycemic action of T-1095 is derived from the enhancement of urinary glucose disposal. Although food intake was not changed in the T-1095-treated rats, the body weight gain was retarded. T-1095 treatment partially ameliorated oral glucose tolerance but not the impaired glucose-induced insulin secretion. Homeostasis model assessment (HOMA) indicated the existence of insulin resistance in GK rats and a significant restoration by T 1095-treatment. There was a reduction of the thermal response in tail-flick testing following long-term hyperglycemia (diabetic neuropathy). Treatment of T 1095 significantly prevented the development of diabetic neuropathy in male GK rats. Sustained improvement of hyperglycemia and prevention of diabetic neuropathy by the T-1095-treatment provide further support the use of SGLT inhibitors for the treatment of diabetes. PMID- 15792834 TI - Inhibitory effect of vitamin E on proinflammatory cytokines-and endotoxin-induced nitric oxide release in alveolar macrophages. AB - Nitric oxide is thought to be an important modulator of various functions in normal and inflamed airways. In the present study, we evaluated the effects of high vitamin E (250 mg and 1250 mg alpha-tocopheryl acetate (TA)/kg diet/10 days) on nitric oxide (NO(.)) release by alveolar macrophages (AMs) in response to lipopolysaccharide (LPS), interleukin-1beta (IL-1beta) and tumor necrosis factor (TNF-alpha). LPS and IL-1beta treatment (1-10 microg/ml) enhanced NO(.) release in AMs from control animals fed on 50 mg vitamin E/kg diet in a concentration dependent manner. However, this enhancement of NO(.) was attenuated in the AMs of animals fed with 250 mg or 1250 mg vitamin E/kg diet. TNF-alpha had no effect in eliciting the release of NO(.) in AMs obtained either from control or from hyper vitamin E fed animals. Further, LPS (1-10 microg/ml) enhanced the inducible nitric oxide synthase (iNOS) activity of AMs of control group and TA-fed animals almost to equal extent. Similarly, LPS-induced formation of N-nitrosamine (N nitroso-L-[(14)C]-proline) in AMs of control and TA-supplemented animals were not different statistically. On the other hand, in vitro addition of vitamin E (200 microM) in AMs of control animals, when triggered with 10 microg LPS/ml, caused a significant decrease in N-nitroso-L-[(14)C]-proline formation. It seems that high doses of TA in diet may play a role in reducing the lipopolysaccharide and proinflammatory cytokines-induced NO(.)-mediated damage by AMs. PMID- 15792835 TI - Regulation of aminopeptidase N (EC 3.4.11.2; APN; CD13) by interferon-gamma on the HL-60 cell line. AB - Membrane-bound peptidases play important roles in the regulation of local concentrations of various signalling peptides such as the growth factors, hormones, chemokines and cytokines. That is accomplished by means of their enzyme activity. Recently, membrane-bound peptidases have also been shown to act as receptors, receiving signals from as yet undefined ligands and transducing them into the cell interior. By using either or both of these mechanisms, peptidases interact with fundamental cellular functions: growth, differentiation, activation and death. This study addressed the effects of a T-cell derived cytokine, interferon-gamma (IFN-gamma) on the activity of aminopeptidase N (APN), an ectoenzyme processing several signal peptides. Cells of a myelo-monocytic cell line HL-60 were used as a model system, and APN was assayed at the levels of mRNA, its membrane marker CD13, and the enzyme activity. Regulation of CD13/APN by IFN-gamma was found at all three levels. The direction of regulation was time dependent: an initial down-regulation seen 24 and 48 hrs after the onset of treatment with IFN-gamma was replaced by an up-regulation after 72 and/or 96 hrs. Up-regulation of CD13/APN observed after 96 hrs was preceded by an up-regulation of APN mRNA reaching its maximum after 72 hrs. The IFN-gamma-induced regulation of APN was due to membrane aminopeptidase N, since it could be completely abrogated by an APN blocking antibody WM-15. The delayed up-regulation of CD13/APN (observed after 72 and/or 96 hrs), required de novo protein synthesis as it could be abrogated by cycloheximide, an inhibitor of protein synthesis. Possible role of endogenous (IFN-gamma-induced) TGF-beta in mediating CD13/APN up regulation could be excluded, since no TGF-beta was found in supernatants of IFN gamma treated HL-60 cells. Thus, our data show regulation of CD13/APN on cells of myelo-monocytic origin by a T-cell derived cytokine, IFN-gamma. A similar mechanism might play a role in inflammation. PMID- 15792836 TI - Osteoblastic activity and estrogenic response in the regenerating scale of goldfish, a good model of osteogenesis. AB - Osteogenesis in the teleost was morphologically observed using regenerating scales of goldfish. Histological observations indicated that osteoblasts around the regenerating scales on days 7 to 10 were greater in size and number than those at other stages. Therefore, further experiments were carried out to examine the activity of osteoblasts in the regenerating period. To quantify their osteoblastic activities, scales on the left side of the body were taken, and the regenerating scales were then used to measure the activities of alkaline phosphatase (ALP), a marker of osteoblasts, on days 7, 10, and 15. The ontogenic scales on the right side of the body were also collected and used to measure ALP activity on the same days. Osteoblasts at all stages of regenerating scales were more active than those in the remaining ontogenic scales. The regenerating scales on day 10 had the highest activity. Furthermore, we found that estrogen receptor (ER) mRNA was expressed in the regenerating scales because estrogen participates in osteoblastic growth and differentiation in mammals. Therefore, using a scale culture system reported previously, the estrogenic response was examined in the ontogenic and regenerating scales on day 10. The reactivity was much higher in regenerating scales, although estrogen treatment significantly activated the osteoblastic activities in both scales. We are the first to demonstrate that ER is expressed in regenerating scales and that estrogen participates in osteogenesis as it does in mammalian bone. Our findings strongly suggest that regenerating scales can be used as a model of osteogenesis in vertebrates. PMID- 15792837 TI - A Src family inhibitor (PP1) potentiates tumor-suppressive effect of connexin 32 gene in renal cancer cells. AB - Connexin (Cx) genes exert negative growth effects on tumor cells with certain cell specificity. We have recently reported that Cx32 acts as a tumor suppressor gene in renal cancer cells due to the inhibition of Src-dependent signaling. In line with the previous study, here we examined if a Src family inhibitor (PP1) could potentiate tumor-suppressive effect of Cx32 in Caki-2 cell from human renal cell carcinoma. In order to clarify the potentialization of PP1, using Cx32 transfected Caki-2 cells and mock-transfected Caki-2 cells, we estimated difference in cytotoxic effect of PP1 on the two cell clones in vitro as well as in vivo. PP1 showed more cytotoxic effect on Caki-2 cells having Cx32 positive expression than that of Cx32 negative expression at lower doses. This potentialization was also observed in xenograft model of nude mice. The potentialization of the effect mainly depended on the induction of apoptosis but not the control of cell growth. In conjugation with this event, the reduction of anti-apoptotic molecules (Bcl-2 and Bcl-xL) was caused by the combination of Cx32 expression and PP1 treatment in Caki-2 cells. These results suggest that PP1 potentiates tumor-suppressive effect of connexin 32 gene in renal cancer cells through the reduction of anti-apoptotic molecules. PMID- 15792838 TI - Effects of different fractions of a red wine non-alcoholic extract on ischemia reperfusion injury. AB - We have recently demonstrated the cardioprotective effects of a non-alcoholic extract of Argentinian red wine (RWE) on ischemia-reperfusion injury. The aim of the present study was to assess the relative contribution of four phenolic fractions separated from RWE by liquid/liquid extraction with solvents of decreasing hydrophobicity, to the myocardial protection achieved by the original extract. Isovolumic perfused rat hearts treated with each fraction 10 min before ischemia and the first 10 min of reperfusion were submitted to a 20-min global ischemic period followed by 30 min of reperfusion. The treatment with the fraction rich in polymeric proanthocyanidins (fraction IV = aqueous residue) significantly improved the postischemic recovery of left ventricular developed pressure (LVDP) and +dP/dt (max) (111 +/- 5% and 117 +/- 6% vs 61 +/- 4%, 62 +/- 5% , respectively, detected in control hearts) and abolished the increase of left ventricular end diastolic pressure (LVEDP) (8 +/- 2 mmHg vs 42 +/- 4 mmHg in untreated hearts). However, the fraction rich in anthocyanins (III: butanol) elicited a cardioprotective action weaker than the original extract. On the other hand, the representative of either resveratrol or flavan-3-ols and flavonols (fractions I and II) failed to induce this type of response. LDH release and TBARS concentration were significantly lowered after treatment with fraction IV alone. These data show that the fraction rich in polymeric proanthocyanidins exerts a protective effect against myocardial alterations derived from ischemia and reperfusion comparable to the original RWE. This beneficial effect can be correlated to the ability of that fraction to attenuate the degree of lipid peroxidation. PMID- 15792839 TI - Sex differences in expression of calcium-handling proteins and beta-adrenergic receptors in rat heart ventricle. AB - Human studies reveal sex differences in myocardial function as well as in the incidence and manifestation of heart disease. Myocellular Ca(2+) cycling regulates normal contractile function; whereas cardiac dysfunction in heart failure has been associated with alterations in Ca(2+)-handling proteins. Beta adrenergic receptor (beta-AR) signaling regulates activity of several Ca(2+) handling proteins and alterations in beta-AR signaling are associated with heart disease. This study examines sex differences in expression of beta(1)-AR, beta(2) AR, and Ca(2+)-handling proteins including: L-type calcium channel (Ca(v)1.2) , ryanodine calcium-release channels (RyR), sarcoplasmic reticular Ca(2+) ATPase (SERCA2), phospholamban (PLB) and Na(+)-Ca(2+) exchange protein (NCX) in healthy hearts from male and female Sprague-Dawley rats. Protein levels were examined using Western blot analysis. Abundance of mRNA was determined by real time RT-PCR normalized to abundance of GAPDH mRNA. Contraction parameters were measured in right ventricular papillary muscle in the presence and absence of isoproterenol. Results demonstrate that female ventricle has significantly higher levels of Ca(v)1.2, RyR, and NCX protein compared to males. Messenger RNA abundance for RyR, and NCX protein was significantly higher in females whereas Ca(v)1.2 mRNA was higher in males. No differences were detected in beta-ARs, SERCA2 or PLB. Female right papillary muscle had a faster maximal rate of force development and decline (+/- dF/dt). There were no sex differences in response to isoproterenol. Results show significant sex differences in expression of key ventricular Ca(2+) handling proteins that are associated with small functional differences in +/- dF/dt. Further studies will determine whether differences in the abundance of these key proteins play a role in sex disparities in the incidence and manifestation of heart disease. PMID- 15792840 TI - Sympathetic innervation of ciliary muscle and oculomotor function in emmetropic and myopic young adults. AB - PURPOSE: Evidence exists for an additional inhibitory accommodative control system mediated by the sympathetic branch of the autonomic nervous system (ANS). This work aims to show the relative prevalence of sympathetic inhibition in young emmetropic and myopic adults, and to evaluate the effect of sympathetic facility on accommodative and oculomotor function. METHODS: Profiling of ciliary muscle innervation was carried out in 58 young adult subjects (30 emmetropes, 14 early onset myopes, 14 late onset myopes) by examining post-task open-loop accommodation responses, recorded continuously by a modified open-view infrared optometer. Measurements of amplitude of accommodation, tonic accommodation, accommodative lag at near, AC/A ratio, and heterophoria at distance and near were made to establish a profile of oculomotor function. RESULTS: Evidence of sympathetic inhibitory facility in ciliary smooth muscle was observed in 27% of emmetropes, 21% of early-onset myopes and 29% of late-onset myopes. Twenty-six percent of all subjects demonstrated access to sympathetic facility. Closed-loop oculomotor function did not differ significantly between subjects with sympathetic facility, and those with sympathetic deficit. CONCLUSIONS: Emmetropic and myopic groups cannot be distinguished in terms of the relative proportions having access to sympathetic inhibition. Presence of sympathetic innervation does not have a significant effect on accommodative function under closed-loop viewing conditions. PMID- 15792841 TI - Retinal properties and potential of the adult mammalian ciliary epithelium stem cells. AB - The ciliary epithelium (CE) in the adult mammalian eye harbors a mitotic quiescent population of neural stem cells. Here we have compared the cellular and molecular properties of CE stem cells and populations of retinal progenitors that define the early and late stages of histogenesis. The CE stem cells and retinal progenitors proliferate in the presence of mitogens and share the expression of universal neural and retinal progenitor markers. However, the expression of the majority of retinal progenitor markers (e.g., Chx10) is transient in the former when compared to the latter, in vitro. They are similar to early than late retinal progenitors in their proliferative response to FGF2 and/or EGF. Analysis of the differentiation potential of CE stem cells shows that they are capable of generating both early (e.g., retinal ganglion cells) and late (e.g., rod photoreceptors) born retinal neurons. However, under identical differentiation conditions, i.e., in the presence of 1% FBS, they generate more early-born retinal neurons than late-born retinal neurons showing a preference for generating early retinal neurons. Transcription profiling of these cells and retinal progenitors demonstrate that they share approximately 80% of the expressed genes. The CE stem cells have more unique genes in common with early retinal progenitors than late retinal progenitors. Both proliferative/differential potential and transcription profiles suggest that CE stem cells may be a residual population of stem cells of optic neuroepithelium, representing a stage antecedent to retinal progenitors. PMID- 15792842 TI - Ocular compensation for alternating myopic and hyperopic defocus. AB - During development, the eye grows under visual feedback control, as shown by its compensating for defocus imposed by spectacle lenses. Under normal conditions the sign and magnitude of defocus vary with viewing distance, accommodative status and other factors. To explore how periods of myopic and hyperopic defocus are integrated over time we presented rapidly alternating episodes of myopic and hyperopic defocus by sequentially illuminating a nearby scrim and the wall beyond it to chick eyes wearing lenses that put the far point between the two surfaces. We found that equal periods of myopic and hyperopic defocus generally led to compensatory hyperopia, showing that myopic defocus had a disproportionate effect. Furthermore, the degree of hyperopia depended on the frequency of alternation: low frequencies (1 cycle/30 min) resulted in more hyperopia, whereas at high frequencies (1 cycle/s) the myopic and hyperopic defocus nearly cancelled each other. If similar temporal integration effects apply to humans, they may help explain why brief accommodation events may not influence lens-compensation and why a child's total reading time may be a poor predictor of myopic progression. PMID- 15792843 TI - Absolute travel distance from optic flow. AB - Optic flow fields provide rich information about the observer's self-motion. Besides estimation of the direction of self-motion human observers are also able to discriminate the travel distances of two self-motion simulations. Recent studies have shown that observers estimate the simulated ego velocity of the self motion simulation and integrate it over time. Thus, observers use a 3-D percept of the ego motion through the environment. In the present work we ask if human observers are able to use this 3-D percept of the motion simulation to build up an internal representation of travel distance and indicate it in a static scene. We visually simulated self-motion in different virtual environments and asked subjects to indicate the perceived distances in terms of static virtual intervals on the ground. The results show that human observers possess a static distance gauge, but that they undershoot the travel distances for short motion simulations. In further experiments we changed the modality of the distance indication but the undershoot in distance estimation remained. This suggests that the undershoot is linked to the perception of the optic flow field. PMID- 15792844 TI - Parafoveal-on-foveal effects on eye movements in text reading: does an extra space make a difference? AB - Schiepers proposed that in text reading, the currently fixated word and the next word are processed in parallel but with a time delay of 90 ms per degree of eccentricity. In his model, the benefit of seeing the upcoming word is due to the fact that the parafoveal information from fixation n is combined with the foveal information from fixation n+1 to boost word recognition, at least when the fixation on word n is of an optimal duration (between 210 and 270 ms). We tested this assumption by adding an extra blank space between the foveal and the parafoveal word. According to the model, this should result in a 30 ms longer processing time for the foveal word. However, reading time was shorter for a word followed by a double space than for a word followed by a single space. An effect of parafoveal word length was also observed with a longer word in the parafoveal leading to shorter fixation times on the foveal word. Implications of these low level parafoveal-on-foveal effects are discussed. PMID- 15792845 TI - At first sight: a high-level pop out effect for faces. AB - To determine the nature of face perception, several studies used the visual search paradigm, whereby subjects detect an odd target among distractors. When detection reaction time is set-size independent, the odd element is said to "pop out", reflecting a basic mechanism or map for the relevant feature. A number of previous studies suggested that schematic faces do not pop out. We show that natural face stimuli do pop out among assorted non-face objects. Animal faces, on the other hand, do not pop out from among the same assorted non-face objects. In addition, search for a face among distractors of another object category is easier than the reverse search, and face search is mediated by holistic face characteristics, rather than by face parts. Our results indicate that the association of pop out with elementary features and lower cortical areas may be incorrect. Instead, face search, and indeed all feature search, may reflect high level activity with generalization over spatial and other property details. PMID- 15792846 TI - Laminar cortical dynamics of 3D surface perception: stratification, transparency, and neon color spreading. AB - The 3D LAMINART neural model is developed to explain how the visual cortex gives rise to 3D percepts of stratification, transparency, and neon color spreading in response to 2D pictures and 3D scenes. Such percepts are sensitive to whether contiguous image regions have the same contrast polarity and ocularity. The model predicts how like-polarity competition at V1 simple cells in layer 4 may cause these percepts when it interacts with other boundary and surface processes in V1, V2, and V4. The model also explains how: the Metelli Rules cause transparent percepts, bistable transparency percepts arise, and attention influences transparency reversal. PMID- 15792847 TI - Spectral and directional reflectance of the fovea in diabetes mellitus: photoreceptor integrity, macular pigment and lens. AB - Our aim was to assess the integrity of the photoreceptors in the fovea, and to measure the optical density of the macular pigment and the eye lens in patients with diabetes mellitus, and to compare the results with those of a group of healthy subjects. The directional and spectral properties of the light reflected from a 1.9 deg field centered on the fovea were measured simultaneously, in a single one second flash, with the Foveal Reflection Analyzer. The directional characteristics, i.e., the optical Stiles-Crawford effect, provided information on the integrity of the foveal photoreceptors. Model analysis of the spectral reflectance yielded optical densities of the macular pigment and the lens. The amplitude of the directional reflectance in diabetic eyes was significantly lower compared to controls (P<0.001). This indicates that the integrity of the photoreceptors in the fovea was altered in diabetics. Surprisingly, the directionality (a measure for the peakedness) was similar in diabetics and controls (P=0.3). The density of macular pigment was not different from that in controls (P=0.3). The optical density of the lens increased with age in both groups, but the rate of increase was larger in the diabetics (P<0.05). Possibly, the lens optical density increasing at a higher rate with age reflects changes preceding cataract formation. PMID- 15792848 TI - Alteration of the perceived path of a non-pursued target during smooth pursuit: analysis by a neural network model. AB - During pursuit of a circularly moving target, the perceived movement of a second circularly moving target is altered. The perceived movement of the non-pursued target is different from both its real movement path and its retinal path. In the present paper this phenomenon is studied using a physiologically based neural network model. Simulation results were compared to psychophysical findings in human subjects. Model simulations enabled us to suggest an explanation for this phenomenon in terms of underlying physiological mechanisms and to estimate the contribution of the efferent eye-movement signal to the perceptual process. PMID- 15792849 TI - Spatio-temporal requirements for direction selectivity in area 18 and PMLS complex cells. AB - The spatio-temporal requirements for direction selectivity were studied in two extrastriate motion processing areas in the cat, area 18 and the posteromedial lateral suprasylvian cortex (PMLS). Direction, velocity and pixel size of random pixel arrays (RPA) were adjusted for each neuron and direction selectivity was measured as a function of step size and delay for a given optimal velocity. A subset of direction selective complex cells in area 18 was tuned to intermediate step size and delay combinations rather than the smoothest motion (band-pass cells). Other area 18 complex cells responded best to the smallest value of step size and delay (low-pass cells). Tuning varied with the pixel size of the RPA. Cells with tuning for smaller pixels favoured a preference for non-smooth motion. Area 18 cells with lower spatial resolution showed larger optimal and maximal step sizes. For a subset of the cells in area 18, we measured direction selectivity for extensive step-delay combinations, covering multiple velocities. Results showed that most cells were tuned to narrow range of step-delay combinations, and that the optimal step size was independent of temporal delay. Direction selective complex cells in PMLS were tuned to larger pixel sizes than those in area 18, although the distributions did overlap. In contrast to area 18, PMLS cells preferred the smoothest motion, irrespective of RPA pixel size. PMID- 15792850 TI - Thought-action fusion: review of the literature and future directions. AB - Thought-action fusion (TAF) is the tendency for individuals to assume that certain thoughts either imply the immorality of their character or increase the likelihood of catastrophic events. The burgeoning literature on TAF is reviewed. It is not clear whether TAF refers to a specific appraisal style, a more enduring belief, or a combination of both. Inconsistent definitions of magical thinking have hindered better understanding of the relationship between TAF and magical thinking. Much work remains to be done to improve assessment and measurement of TAF. TAF is associated with tendencies towards obsessive-compulsive disorder (OCD) and may contribute to its symptoms. However, the literature investigating TAF and other variables implicated in OCD remains inconclusive. It is suggested that TAF is not specific to OCD, but also prevalent in other anxiety disorders. TAF appears to be moderately related to depressive symptoms and a similar bias may contribute to preoccupations in eating disorders. TAF is also associated with the presence of psychological disorders in children and adolescents. Educational and cognitive therapy approaches to reduce TAF and consequent symptoms are discussed, and suggestions for further research made. PMID- 15792851 TI - Voluntary hyperventilation in the treatment of panic disorder--functions of hyperventilation, their implications for breathing training, and recommendations for standardization. AB - Hyperventilation has numerous theoretical and empirical links to anxiety and panic. Voluntary hyperventilation (VH) tests have been applied experimentally to understand psychological and physiological mechanisms that produce and maintain anxiety, and therapeutically in the treatment of anxiety disorders. From the theoretical perspective of hyperventilation theories of anxiety, VH is useful diagnostically to the clinician and educationally to the patient. From the theoretical perspective of cognitive-behavior therapy, VH is a way to expose patients with panic disorder to sensations associated with panic and to activate catastrophic cognitions that need restructuring. Here we review panic disorder treatment studies using breathing training that have included VH. We differentiate the roles of VH in diagnosis, education about symptoms, training of breathing strategies, interoceptive exposure, and outcome measurement--discussing methodological issues specific to these roles and VH test reliability and validity. We propose how VH procedures might be standardized in future studies. PMID- 15792853 TI - Potential roles of parental self-efficacy in parent and child adjustment: a review. AB - This review examines the potential roles of parental self-efficacy (PSE) in parent and child adjustment and the role of parental cognitions in understanding behaviors and emotions within families. The areas in this review include parental competence and psychological functioning, as well as child behaviors, socio emotional adjustment, school achievement, and maltreatment. There is strong evidence linking PSE to parental competence, and more modest linkage to parental psychological functioning. Some findings suggest that PSE impacts child adjustment directly but also indirectly via parenting practices and behaviors. Although the role of PSE likely varies across parents, children, and cultural contextual factors, its influence cannot be overlooked as a possible predictor of parental competence and child functioning, or perhaps an indicator of risk. PSE may also be an appropriate target for prevention and intervention efforts. Limitations in the literature include measurement problems, variability in conceptualizations and definitions of the construct, and the lack of research exploring causality. Future research should focus on clarifying the measurement of PSE, studying potential bias in self-report of PSE, and utilizing experimental and longitudinal designs to untangle the issues of causal direction and potential transactional processes. PMID- 15792854 TI - Psychosocial impact of parental cancer in adulthood: a conceptual and empirical review. AB - This article reviews recent literature on the psychosocial aspects of parental cancer in adulthood. Overall, studies have shown that a sizable minority of adult children of cancer patients experience psychological distress in terms of anxiety, depression, and posttraumatic stress symptoms. Most research to date has focused on the first-degree female relatives of breast cancer patients, many of whom are daughters. Although distress reactions in this population are well documented, restrictive assumptions reflecting a vulnerability/deficit model of women limit scientific progress. In addition, the critical role of grown offspring caregivers of cancer patients deserves further investigation. Future research should examine the relations between intrapsychic and contextual factors that may influence psychological adjustment to parental cancer. PMID- 15792855 TI - Thermal denaturation of the BRCT tandem repeat region of human tumour suppressor gene product BRCA1. AB - Reduced stability of the tandem BRCT domains of human BReast CAncer 1 (BRCA1) due to missense mutations may be critical for loss of function in DNA repair and damage-induced checkpoint control. In the present thermal denaturation study of the BRCA1 BRCT region, high-precision differential scanning calorimetry (DSC) and circular dichroism (CD) spectroscopy provide evidence for the existence of a denatured state that is structurally very similar to the native. Consistency between theoretical structure-based estimates of the enthalpy (DeltaH) and heat capacity change (DeltaCp) and the calorimetric results is obtained when considering partial thermal unfolding contained in the region of the conserved hydrophobic pocket formed at the interface of the two BRCT repeats. The structural integrity of this region has been shown to be crucial for the interaction of BRCA1 with phosphorylated peptides. In addition, cancer-causing missense mutations located at the inter-BRCT-repeat interface have been linked to the destabilization of the tandem BRCT structure. PMID- 15792856 TI - NMR spectroscopic, mass spectroscopic, X-ray crystallographic, and theoretical studies of molecular mechanics of natural products: farformolide B and sesamin. AB - Two natural products, farformolide B and sesamin were isolated from Farfugium japonicum and Cinnamomum kanehirae, respectively. The structures of the two natural products, including their relative stereochemistry, were elucidated using spectroscopic data and theoretical calculations. The molecule 1 (farformolide B) is newly recognized by X-ray crystallography. The two compounds were also investigated by a theoretical analysis using the B3LYP/6-31G* method of the Gaussian 03 package program. The theoretical results were supplemented by experimental data to determine the optimal geometric structures of the two compounds. The calculated molecular mechanics were found to compare well with the experimental data. Several important thermodynamic properties of the two products, including ionization potentials, highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) energies, energy gaps, heat of formation, atomization energies, and vibration frequencies, were also calculated. The study also provided a good understanding of the stereochemical structure and thermodynamic properties of the two molecules. PMID- 15792857 TI - Electrochemical studies of danthron and the DNA-danthron interaction. AB - Danthron is an important natural occurring component in laxative drugs. In this paper, electrochemical investigation of danthron and its interaction with DNA is reported. Via the electrochemical approach assisted by ultraviolet-visible (UV Vis) spectroscopy, we have proved that danthron intercalates into DNA strands forming some nonelectroactive complexes, which results in the decrease of redox peak currents of danthron. In addition, the decrease of the peak currents is proportional to the concentration of DNA. The difference between the interaction of danthron with double-stranded DNA (dsDNA) and with single-stranded DNA (ssDNA) has also been studied. This character implies the potential of danthron to discriminate dsDNA and ssDNA. PMID- 15792852 TI - Distinctions between bipolar and unipolar depression. AB - This is a review of the studies comparing unipolar and bipolar depression, with focus on the course, symptomatology, neurobiology, and psychosocial literatures. These are reviewed with one question in mind: does the evidence support diagnosing bipolar and unipolar depressions as the same disorder or different? The current nomenclature of bipolar and unipolar disorders has resulted in research that compares these disorders as a whole, without considering depression separately from mania within bipolar disorder. Future research should investigate two broad categories of depression and mania as separate disease processes that are highly comorbid. PMID- 15792858 TI - Spectroscopic markers of the T<-->R quaternary transition in human hemoglobin. AB - In this work, we use a sol-gel protocol to trap and compare the R and T quaternary states of both the deoxygenated (deoxyHb) and carbonmonoxide (HbCO) derivatives of human hemoglobin. The near infrared optical absorption band III and the infrared CO stretching band are used to detect the effect of quaternary structure on the spectral properties of deoxyHb and HbCO; comparison with myoglobin allows for an assessment of tertiary and quaternary contributions to the measured band shifts. The R<-->T transition is shown to cause a blue shift of the band III by approximately 35 cm(-1) for deoxyHb and a red shift of the CO stretching band by only approximately 0.3 cm(-1) for HbCO. This clearly shows that quaternary structure changes are transmitted to the heme pocket and that effects on deoxyHb are much larger than on HbCO, at least as far as the band energies are concerned. Experiments performed in the ample temperature interval of 300-10K show that the above quaternary structure effects are "static" and do not influence the dynamic properties of the heme pocket, at least as probed by the temperature dependence of band III and of the CO stretching band. The availability of quaternary structure sensitive spectroscopic markers and the quantitative measurement of the quaternary structure contribution to band shifts will be of considerable help in the analysis of flash-photolysis experiments on hemoglobin. Moreover, it will enable one to characterize the dynamic properties of functionally relevant hemoglobin intermediates and to study the kinetics of both the T-->R and R-->T quaternary transitions through time-resolved spectroscopy. PMID- 15792859 TI - Dimethyl-pepep: a DNA probe in two-photon excitation cellular imaging. AB - Dimethyl-pepep (D-pepep), a newly developed and very efficient two-photon absorber, has been tested here for two-photon excitation (TPE) cellular imaging. The spectral characteristics of the dye following one-photon excitation (OPE) and TPE (excitation and emission spectra, fluorescence lifetime, molecular brightness, saturation intensity) are reported. In vitro interaction studies with biomolecules show that dimethyl-pepep has a large affinity for DNA. A comparison with a widely used DNA stainer, 4-6-diamidino-2-phenylindole (DAPI) bound to DNA shows that the D-pepep brightness is one order of magnitude higher than that of DAPI, making this dye suitable for microscopy and imaging applications. TPE images taken from double-stained yeast Saccharomyces cerevisiae cells have revealed that D-pepep localizes mainly in the nucleus, similarly to DAPI, and in mitochondria, although to a minor extent. Preliminary tests have shown that the dye cellular toxicity is negligible. PMID- 15792860 TI - Ligand-linked stability of mutants of the C-domain of calmodulin. AB - There is a necessary energetic linkage between ligand binding and stability in biological molecules. The critical glutamate in Site 4 was mutated to create two mutants of the C-domain of calmodulin yielding E140D and E140Q. These proteins were stably folded in the absence of calcium, but had dramatically impaired binding of calcium. We determined the stability of the mutant proteins in the absence and presence of calcium using urea-induced unfolding monitored by circular dichroism (CD) spectroscopy. These calcium-dependent unfolding curves were fit to models that allowed for linkage of stability to binding of a single calcium ion to the native and unfolded states. Simultaneous analysis of the unfolding profiles for each mutant yielded estimates for calcium-binding constants that were consistent with results from direct titrations monitored by fluorescence. Binding to the unfolded state was not an important energetic contributor to the ligand-linked stability of these mutants. PMID- 15792861 TI - Quantification of lipid bilayer effective microviscosity and fluidity effect induced by propofol. AB - Electron spin resonance (ESR) spectroscopy with nitroxide spin probes was used as a method to probe the liposome microenvironments. The effective microviscosities have been determined from the calibration of the ESR spectra of the probes in solvent mixtures of known viscosities. In the first time, by measuring ESR order parameter (S) and correlation time (tau(c)) of stearic spin probes, we have been able to quantify the value of effective microviscosity at different depths inside the liposome membrane. At room temperature, local microviscosities measured in dimyristoyl-l-alpha phosphatidylcholine (DMPC) liposome membrane at the different depths of 7.8, 16.95, and 27.7 A were 222.53, 64.09, and 62.56 cP, respectively. In the gel state (10 degrees C), those microviscosity values increased to 472.56, 370.61, and 243.37 cP. In a second time, we have applied this technique to determine the modifications in membrane microviscosity induced by 2,6-diisopropyl phenol (propofol; PPF), an anaesthetic agent extensively used in clinical practice. Propofol is characterized by a unique phenolic structure, absent in the other conventional anaesthetics. Indeed, given its lipophilic property, propofol is presumed to penetrate into and interact with membrane lipids and hence to induce changes in membrane fluidity. Incorporation of propofol into dimyristoyl-l alpha phosphatidylcholine liposomes above the phase-transition temperature (23.9 degrees C) did not change microviscosity. At 10 degrees C, an increase of propofol concentration from 0 to 1.0 x 10(-2) M for a constant lipid concentration mainly induced a decrease in microviscosity. This fluidity effect of propofol has been qualitatively confirmed using merocyanine 540 (MC540) as lipid packing probe. Above 10(-2) M propofol, no further decrease in microviscosity was observed, and the microviscosity at the studied depths (7.8, 16.95, and 27.7 A) amounted 260.21, 123.87, and 102.27 cP, respectively. The concentration 10(-2) M was identified as the saturation limit of propofol in dimyristoyl-l-alpha phosphatidylcholine liposomes. PMID- 15792862 TI - Solvent accessibility in native and isolated domain environments: general features and implications to interface predictability. AB - A non-redundant database of 4536 structural domains, comprising more than 790,000 residues, has been used for the calculation of their solvent accessibility in the native protein environment and then in the isolated domain environment. Nearly 140,000 (18%) residues showed a change in accessible surface area in the above two conditions. General features of this change under these two circumstances have been pointed out. Propensities of these interfacing amino acid residues have been calculated and their variation for different secondary structure types has been analyzed. Actual amount of surface area lost by different secondary structures is higher in the case of helix and strands compared to coil and other conformations. Overall change in surface area in hydrophobic and uncharged residues is higher than that in charged residues. An attempt has been made to know the predictability of interface residues from sequence environments. This analysis and prediction results have significant implications towards determining interacting residues in proteins and for the prediction of protein-protein, protein-ligand, protein-DNA and similar interactions. PMID- 15792863 TI - Tryptophan interactions with glycerol/water and trehalose/sucrose cryosolvents: infrared and fluorescence spectroscopy and ab initio calculations. AB - In order to correlate how the solvent affects emission properties of tryptophan, the fluorescence and phosphorescence emission spectra of tryptophan and indole model compounds were compared for solid sugar glass (trehalose/sucrose) matrix and glycerol/water solution and under the same conditions, these matrices were examined by infrared spectroscopy. Temperature was varied from 290 to 12 K. In sugar glass, the fluorescence and phosphorescence emission spectra are constant over this temperature range and the fluorescence remains red shifted; these results are consistent with the static interaction of OH groups with tryptophan in the sugar glass. In sugar glass containing water, the water retains mobility over the entire temperature range as indicated by the HOH infrared bending frequency. The fluorescence of tryptophan in glycerol/water shifts to the blue as temperature decreases and the frequency change of the absorption of the HOH bend mode is larger than in the sugar glass. These results suggest rearrangement of glycerol and water molecules over the entire temperature change. Shifts in the fluorescence emission maximum of indole and tryptophan were relatively larger than shifts for the phosphorescence emission-as expected for the relatively smaller excited triplet state dipole for tryptophan. The fluorescence emission of tryptophan in glycerol/water at low temperature has maxima at 312, 313, and 316 nm at pH 1.4, 7.0, and 10.6, respectively. The spectral shifts are interpreted to be an indication of a charge, or Stark phenomena, effect on the excited state molecule, as supported by ab initio calculations. To check whether the amino acid remains charged over the temperature range, the infrared spectrum of alanine was monitored over the entire range of temperature. The ratio of infrared absorption characteristic of carboxylate/carbonyl was constant in glycerol/water and sugar glass, which indicates that the charge was retained. Tryptophan buried in proteins, namely calcium parvalbumin from cod and aldolase from rabbit, showed temperature profiles of the fluorescence spectra that were largely independent of the solvent (glycerol/water or sugar glass) and temperature whereas the fluorescence and phosphorescence yields were dependent. The results demonstrate how the rich information found in tryptophan luminescence can provide information on the dipolar nature and dynamics of the matrix. PMID- 15792864 TI - Walker-Warburg syndrome and limb girdle muscular dystrophy; two sides of the same coin. PMID- 15792865 TI - An autosomal recessive limb girdle muscular dystrophy (LGMD2) with mild mental retardation is allelic to Walker-Warburg syndrome (WWS) caused by a mutation in the POMT1 gene. AB - Mutations of the protein O-mannosyltransferase (POMT1) gene affect glycosylation of alpha-dystroglycan, leading to Walker-Warburg syndrome, a lethal disorder in early life with severe congenital muscular dystrophy, and brain and eye malformations. Recently, we described a novel form of recessive limb girdle muscular dystrophy with mild mental retardation, associated with an abnormal alpha-dystroglycan pattern in the muscle, suggesting a glycosylation defect. Here, we present evidence that this distinct phenotype results from a common mutation (A200P) in the POMT1 gene. Our findings further expand the phenotype of glycosylation disorders linked to POMT1 mutations. Furthermore, the A200P mutation is part of a conserved core haplotype, indicating an ancestral founder mutation. PMID- 15792866 TI - Mitochondrial encephalomyopathies: an update. AB - A genetic classification of the mitochondrial encephalomyopathies includes disorders due to defects of mitochondrial DNA (mtDNA) and disorders due to defects of nuclear DNA (nDNA). Recent progress in mtDNA-related diseases includes: (i) new pathogenic mutations in protein-coding genes, especially those encoding subunits of complex I (ND genes); (ii) the pathogenic nature of homoplasmic mutations, whose expression is regulated by environmental and genetic factors; (iii) increasing interest in the functional and pathophysiological role of haplotypes. Advances in mendelian mitochondrial diseases include: (i) new mutations in genes for complex I subunits; (ii) identification of new mutant ancillary proteins associated with complex IV and complex V deficiencies; (iii) better molecular understanding of disorders due to faulty intergenomic communication, which are associated with multiple mtDNA deletions, mtDNA depletion, or defects of mtDNA translation; (iv) the pathogenic role of alterations of the inner mitochondrial membrane phospholipid components, especially cardiolipin; (v) the emerging importance of defects in mitochondrial motility, fission, or fusion. PMID- 15792867 TI - Temperament and character in patients with classical myotonic dystrophy type 1 (DM-1). AB - This study was designed to investigate personality in classical Myotonic Dystrophy (DM-1). Forty-six patients with DM-1 (25 women and 21 men), 31 healthy controls and 37 subjects in a contrast group, consisting of patients with other muscle disorders (spinal muscular atrophy, facioscapulohumeral dystrophy and limb girdle muscular dystrophy), completed the Temperament and Character Inventory (TCI) (Cloninger, 1994). We aimed to establish whether CTG triplet repeat size correlated with ratings of personality dimensions in the TCI. The DM-1 patients scored significantly higher on the TCI dimension Harm avoidance and lower on Persistence, Self-directedness and Cooperativeness. Signs of a personality disorder were found in 20% of the DM-1 patients. No correlation was found between the number of CTG repeats and scores in the TCI. This study indicates deviant personality in classical DM-1 regarding temperament and character, both in comparison to healthy controls and to patients with other muscle disorders with no known brain disorder. PMID- 15792868 TI - Morphological, clinical and genetic aspects in a family with a novel LAMP-2 gene mutation (Danon disease). AB - A family with several cases of severe cardiomyopathy and moderate myopathy is described, affecting two brothers and their cousin as well as their mothers. One boy died of sudden cardiac arrest at 17 years of age. The two brothers were treated with an implantable defibrillator and their mother died suddenly at 40 years of age. Muscle biopsy in males showed vacuolar myopathy in two cases, and no abnormality on standard staining in the third case. Cardiac biopsies showed hypertrophic and vacuolated fibres. Complete absence of LAMP-2 was demonstrated by immunohistochemistry on the vacuolated skeletal and cardiac muscle, but also on the morphologically normal skeletal muscle. Sequencing of LAMP-2 gene showed a novel S157X mutation in exon 4. Danon disease is a rare and potentially lethal cause of hypertrophic cardiomyopathy. Diagnosis can be made by immunohistochemistry performed on cardiac or muscle biopsy, and confirmed by genetic analysis, which also allows for easy family screening and counselling. PMID- 15792869 TI - Early onset myopathy with a novel mutation in the Selenoprotein N gene (SEPN1). AB - Mutations in SEPN1 have been associated with three autosomal recessive congenital myopathies, including rigid spine muscular dystrophy, multiminicore disease and desmin-related myopathy with Mallory body-like inclusions. These disorders constitute the SEPN1 related myopathies (SEPN-RM). On the basis of clinical and laboratory features compatible with SEPN-RM, we performed mutation analysis of SEPN1 in 11 unrelated patients and found one case with pathogenic mutations. He showed early onset axial muscle weakness and developed scoliosis with respiratory insufficiency. Muscle biopsy showed increased variability of fiber size and slight, focal increase of connective tissue. A few fibers showed mini-core changes. SEPN1 mutation analysis revealed that the patient was a compound heterozygote: a previously described insertion (713-714 insA), and a novel nonsense mutation (R439stop). PMID- 15792870 TI - Muscle MRI in Ullrich congenital muscular dystrophy and Bethlem myopathy. AB - The aim of this study was to evaluate the spectrum of muscle involvement on Magnetic Resonance Imaging (MRI) in patients with collagen VI related disorders. Nineteen patients with genetically confirmed collagen VI related disorders, 10 with Bethlem myopathy and 9 with Ullrich congenital muscular dystrophy (CMD), had muscle MRI of their legs using T1 sequences through calves and thighs. In patients with Bethlem myopathy the vasti muscles appeared to be the most frequently and most strikingly affected thigh muscles, with a rim of abnormal signal at the periphery of each muscle and relative sparing of the central part. Another frequent finding was the presence of a peculiar involvement of the rectus femoris with a central area of abnormal signal within the muscle. Patients with Ullrich CMD had a more diffuse involvement of the thigh muscles with relative sparing of sartorius, gracilis and adductor longus. In 8 of the 9 patients with Ullrich CMD, we also observed the peripheral rim of the vastus lateralis and the central area in the rectus femoris observed in patients with Bethlem myopathy. At calf level the results were more variable but a significant proportion of patients with both Bethlem myopathy (8/10) and Ullrich CMD (6/9) showed a rim of abnormal signal at the periphery of soleus and gastrocnemii. Bethlem myopathy and Ullrich CMD patients have distinct patterns of muscle involvement on MRI with some overlap between the two forms. Our results suggest that muscle MR may be used, as an additional tool, to identify patients with collagen VI related disorders. This information is even more important in the patients with a typical Ullrich CMD clinical phenotype but with normal collagen expression of VI in muscle and/or skin. PMID- 15792871 TI - A novel ANT1 gene mutation with probable germline mosaicism in autosomal dominant progressive external ophthalmoplegia. AB - Only four different mutations in the adenine nucleotide translocator 1 (ANT1) gene have been found in families with progressive external ophthalmoplegia (PEO). We report a novel heterozygous C to A transversion at nucleotide 269 in the ANT1 gene in a German family with PEO, predicted to convert a highly conserved alanine at codon 90 to aspartic acid. The mutation was identified in three siblings with PEO, one of them additionally suffered from schizoaffective disorder. Microsatellite analysis showed that the mutation was dominant and inherited from the mother who did not carry the mutation in blood, indicating germ-line mosaicism. PMID- 15792872 TI - The genes encoding for D4Z4 binding proteins HMGB2, YY1, NCL, and MYOD1 are excluded as candidate genes for FSHD1B. AB - Facioscapulohumeral muscular dystrophy is a disease of skeletal muscle, with symptoms including both facial and shoulder girdle weakness and progression to involve the pelvic girdle and extremities in the majority of cases. For most cases of FSHD, the molecular basis of the disease can be identified as a partial deletion of the D4Z4 repeat array on the end of the long arm of chromosome 4. However, in up to 5% of FSHD families there is no linkage to 4q35. These cases are designated as FSHD1B. Proteins have been identified that bind to the D4Z4 repeats of chromosome 4q35. The genes encoding D4Z4 binding proteins YY1, HMGB2, NCL, and MYOD1 were investigated as candidate genes for FSHD1B. Coding sequences and promoter region were analyzed for HMBG2 and no sequence variations were detected. For YY1, all five exons were analyzed and a polymorphism was detected in both the unaffected and affected populations. In nucleolin (NCL), several SNPs were identified, including a SNP causing the non-synonymous change P515H; however, all polymorphisms either occurred in control samples or were previously reported. A novel polymorphism was also detected in MYOD1, but did not represent a disease-specific variation. These results suggest that HMBG2, YY1, NCL, and MYOD1 are unlikely to represent the genes responsible for FSHD in these families. PMID- 15792873 TI - 129th ENMC International Workshop: Clinical Trials for Chronic Inflammatory Demyelinating Polyradiculoneuropathy and Multifocal Motor Neuropathy, 27th October 2004, Schiphol airport, The Netherlands. PMID- 15792874 TI - Photoinduced electron transfer between cytochrome c and a novel 1,4,5,8 naphthalenetetracarboxylic diimide with amphiphilic character. AB - N-dodecyl-N'-(2-phosphonoethyl)-1,4,5,8-naphthalenetetracarboxylic diimide (DNDI) is a novel naphthalenic diimide with amphiphilic character. DNDI was synthesized through the sequential reaction of 1,4,5,8-naphthalenetetracarboxylic dianhydride, first with dodecylamine and then with 2-aminoethylphosphonic acid. Fluorescence measurements showed that DNDI forms excimers in water at sufficiently high concentrations. The fluorescence quantum yield of DNDI in diluted solutions is sensitive to the polarity of the microenvironment, decreasing as going from water to less polar solvents. This property allowed to monitor the incorporation of DNDI into cetyl trimethyl ammonium bromide (CTAB) micelles, with a binding constant of 1.2x10(4) M-1. UV irradiation (365 nm) of solutions containing DNDI and the redox protein cytochrome c (cyt c) resulted in the reduction of the heme iron from the Fe(III) to the Fe(II) state, a reaction that was inhibited by the incorporation of DNDI into CTAB micelles. DNDI formed host-guest complexes with alpha-cyclodextrin (alpha-CD) through the inclusion of the dodecyl group, resulting in an increased aqueous solubility of the compound. PMID- 15792875 TI - Ab initio model study on acetylcholinesterase catalysis: potential energy surfaces of the proton transfer reactions. AB - Ab initio molecular orbital (MO) and hybrid density functional theory (DFT) calculations have been applied to the initial step of the acylation reaction catalyzed by acetylcholinesterase (AChE), which is the nucleophiric addition of Ser200 in catalytic triads to a neurotransmitter acetylcholine (ACh). We focus our attention mainly on the effects of oxyanion hole and Glu327 on the potential energy surfaces (PESs) for the proton transfer reactions in the catalytic triad Ser200-His440-Glu327. The activation barrier for the addition reaction of Ser200 to ACh was calculated to be 23.4 kcal/mol at the B3LYP/6-31G(d)//HF/3-21G(d) level of theory. The barrier height under the existence of oxyanion hole, namely, Ser200-His440-Glu327-ACh-(oxyanion hole) system, decreased significantly to 14.2 kcal/mol, which is in reasonable agreement with recent experimental value (12.0 kcal/mol). Removal of Glu327 from the catalytic triad caused destabilization of both energy of transition state for the reaction and tetrahedral intermediate (product). PESs calculated for the proton transfer reactions showed that the first proton transfer process is the most important in the stabilization of tetrahedral intermediate complex. The mechanism of addition reaction of ACh was discussed on the basis of theoretical results. PMID- 15792876 TI - Evaluation of phototoxic and photoallergic potentials of 13 compounds by different in vitro and in vivo methods. AB - Phototoxic side effects of pharmaceutical and cosmetic products are of increasing concern for patients, dermatologists and the chemical industry. Moreover, the need of new chemicals and drugs puts pressure on pre-clinical test methods for side effects, especially interactive adverse-effects with UV-light. So, the predictive potential of different established test methods, which are used regularly in our departments in order to detect the phototoxic potential of chemicals, were analyzed. Namely the fibroblast 3T3 test, the photo hen's egg test, a guinea pig test for measuring acute photoreactions, and a modified Local Lymph Node Assay, the Integrated Model for the Differentiation of Skin Reactions. Various agents with different photoreactive potential were tested: quinolones like Bay y 3118, ciprofloxacin, enoxacin, lomefloxacin, moxifloxacin, ofloxacin, sparfloxacin, as well as promethazine, chlorpromazine, 8-methoxypsoralen and olaquindox serving as control. Special emphasis was taken to evaluate the capability of the employed test procedures to predict phototoxic side effects in patients. Following our results, both in vitro assays were useful tools to detect photoirritancy while the photoallergic potentials of tested compounds were exclusively detected by an in vivo assay. As long as no in vitro model for photoallergy is available, the UV-IMDS should be considered to evaluate photoallergic properties of a supposed photoreactive agent. PMID- 15792877 TI - Changes in glutathione and xanthophyll cycle pigments in the high light-stressed lichens Umbilicaria antarctica and Lasallia pustulata. AB - Hydrated thalli of two lichen species--Umbilicaria antarctica and Lasallia pustulata--were exposed to high light (1800 micromol m-2s-1) for 30 min. High light exposure led to a decrease of total glutathione in both species, while de epoxidation state of xanthophyll cycle pigments and non-photochemical quenching increased. In the subsequent recovery, the values of de-epoxidation state of xanthophyll cycle pigments decreased towards initial values. Glutathione (GSH) was resynthetised slowly. In conclusion, zeaxanthin-related protection is probably more involved than GSH-related protection in short-term response to high light stress in U. antarctica and L. pustulata. Faster recovery from photoinhibition in L. pustulata than U. antarctica is mainly due to faster conversion of zeaxanthin to violaxanthin and larger GSH pool of former species. PMID- 15792878 TI - The effect of green laser light irradiation on whole blood platelets. AB - BACKGROUND: Laser light irradiation is assumed to have biostimulating effect in various cell types. However, there is still a lack of information concerning response of blood platelets to laser light irradiation. METHODS: In our study we used flow cytometry to monitor the effect of a green Nd-YAG laser (532 nm, 30 mW) irradiation on platelet activation and the expression of activated GPIIbIIIa glycoprotein complex (fibrinogen receptor) of whole blood platelets stained with fluorolabelled monoclonal antibody PAC-1. Also the formation of platelet microparticles and aggregates in a population of whole blood platelets following such irradiation was evaluated. RESULTS: Effects of laser light on platelet activation and reactivity were significant over a wide range of applied energies (p<0.01). While low and medium laser light energies (18 and 54 J) increased platelet activation, the irradiation with a high-energy laser light (108 J) resulted in depressed platelet reactivity and attenuated platelet response to activators. In addition, laser light irradiation had significant influence on the formation of platelet microparticles in either resting (p<0.05) or ADP-activated (p<0.05) platelets, while no significant effect was observed in collagen activated platelets. On the other hand, laser light irradiation significantly increased the formation of platelet aggregates both in resting (p<0.01) and agonists-activated (p<0.05) platelets. CONCLUSIONS: Our results clearly point that the laser light irradiation of blood platelets can trigger signal transduction, leading to platelet activation, as well as the gradual loss of natural platelet reactivity and platelets' ability to respond to activating agents. PMID- 15792879 TI - Effect of albumin on the photodynamic inactivation of microorganisms by a cationic porphyrin. AB - BACKGROUND: Photodynamic inactivation (PDI) employs visible light and a photosensitizer to inactivate cells. The technique is currently clinically used for the treatment of several malignancies. However, the PDI of microorganisms still remains in the research phase. PURPOSE: To study the effect of human blood plasma and human serum albumin (HSA) on the PDI of Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans. METHODS: PDI experiments were performed using white light (30 mW cm-2) and the cationic 5-phenyl-10,15,20 tris(N-methyl-4-pyridyl)porphyrin chloride (TriP[4]) as photosensitizer. RESULTS: The microorganisms could be successfully photoinactivated by TriP[4] when suspended in phosphate buffered saline (PBS). In this medium, P. aeruginosa was the most resistant microorganism. Changing the suspending medium from PBS to human blood plasma reduced the PDI of all three microorganisms. In human blood plasma C. albicans was the most resistant microorganism. The same results were obtained with 4.5% and 7% HSA/PBS suspensions. CONCLUSIONS: Albumin inhibits the PDI of S. aureus, P. aeruginosa and C. albicans in a dose dependent manner. However, our results are encouraging towards the potential future application of PDI for the treatment of superficial wound infections caused by S. aureus, P. aeruginosa and C. albicans. PMID- 15792880 TI - Photoaddition of thienocoumarin derivatives to DNA: stoichiometry and kinetics of binding. AB - Photoreaction of the 6,9-dimethyl-4-methoxymethyl-2H-thieno[3,2-g]-1-benzopyran-2 one (compound I) and 4-acetoxymethyl-6,9-dimethyl-2H-thieno[3,2-g]-1-benzopyran-2 one (compound II) to DNA was studied. The quantitative evaluation of the photobound molecules was performed by means of inductively coupled plasma atomic emission spectrometry (ICP-AES), exploiting the presence of the sulphur atom inside the tricyclic chromophore. The concurrent estimation of the phosphorus atom, present exclusively in the macromolecule, allowed possible intercalation sites to be identified and their involvement in the photoaddition reaction to be determined. The development of a kinetic model made it possible to discriminate and evaluate the single kinetic events that constitute the overall photoaddition process of I and II to DNA. PMID- 15792881 TI - Ultrafast surface solvation dynamics and functionality of an enzyme alpha chymotrypsin upon interfacial binding to a cationic micelle. AB - In this contribution we report studies on enzymatic activity of alpha chymotrypsin (CHT) upon complexation with cationic cetyltrimethylammonium bromide (CTAB) micelle. With picosecond time resolution, we examined solvation dynamics at the interface of CHT-micelle complex, and rigidity of the binding. We have used 5-(dimethyl amino) naphthalene-1-sulfonyl chloride (dansyl chloride; DC) that is covalently attached to the enzyme at the surface sites. The solvation processes at the surface of CHT in buffer solution are found to be mostly in the sub-50 ps time scale. However, at the interface the solvation correlation function decays with time constant 150 ps (65%) and 500 ps (35%), which is significantly different from those found at the enzyme and micellar surfaces. The binding structure of the enzyme-micelle complex was examined by local orientational motion of the probe DC and compared with the case without micelle. The orientational dynamics of the probe DC in the complex reveals a structural perturbation at the surface sites of CHT upon complexation, consistent with other reported structural studies. We also found possible entanglement of charge transfer dynamics of the probe DC on the measured solvation processes by using time-resolved area normalized emission spectroscopic technique. The interfacial solvation process and complex rigidity elucidate the strong recognition mechanism between CHT and the micelle, which is important to understand the biological function of CHT upon complexation with the micelle. PMID- 15792882 TI - Enhanced ALA-induced fluorescence in hyperparathyroidism. AB - INTRODUCTION: Intraoperative localization of parathyroid glands can be challenging especially in minimally invasive surgery. Fluorescence diagnosis using the photosensitizer aminolevulinic acid (ALA) has been described to identify normal parathyroid glands during experimental bilateral neck exploration. The present study evaluated fluorescence differences between hyperplastic and normal parathyroid glands as a precondition for a clinical application of the technique. MATERIALS AND METHODS: Polycystic kidney disease (PKD) rats with hyperparathyroidism due to hyperplastic parathyroid glands and Wistar rats with normal parathyroid glands were photosensitized by peritoneal lavage with ALA solution. After surgical exposure of thyroid and parathyroid glands the operative site was observed under blue light conditions using the d light system to assess fluorescence characteristics of each tissue. Fluorescence intensities of parathyroid glands and surrounding thyroid tissue were measured by spectrometry. Parathyroid hormone in serum of the rats was determined by enzyme linked immunosorbent assay (ELISA). RESULTS: Observation of the exposed thyroid site showed a subjectively stronger red fluorescence of the parathyroid glands in the PKD rats in comparison to the Wistar rats, whereas thyroid tissue appeared equally fluorescent. In the PKD animals, spectrometric fluorescence intensity was 10 times higher in the parathyroid glands than in the thyroid gland, whereas in the Wistar rats the ratio was 3.2:1. Fluorescence intensity in the parathyroid glands was more than twice in the PKD rats than in the Wistar rats, however slightly lower in the thyroid tissue. ELISA confirmed the pathophysiological change of a hyperparathyroidism with significantly increased serum levels of parathyroid hormone in the PKD rats. DISCUSSION: Hyperparathyroidism enhances ALA induced fluorescence of the parathyroid glands. A combined surgical fluorescence strategy may justify a unilateral, minimally invasive approach in selected patients and serve to improve the capability of the surgeon to safely and efficiently manage parathyroid diseases. PMID- 15792883 TI - Electrophysiological studies on human pain perception. AB - OBJECTIVE: We reviewed the recent progress in electrophysiological studies using electroencephalography (EEG), magnetoencephalography (MEG) and repetitive transcranial magnetic stimulation (rTMS) on human pain perception. METHODS: For recording activities following A delta fiber stimulation relating to first pain, several kinds of lasers such as CO2, Tm:YAG and argon lasers are now widely used. The activity is frequently termed laser evoked potential (LEP), and we reviewed previous basic and clinical reports on LEP. We also introduced our new method, epidermal stimulation (ES), which is useful for recording brain activities by the signals ascending through A delta fibers. For recording activities following C fiber stimulation relating to second pain, several methods have been used but weak CO2 laser stimuli applied to tiny areas of the skin were recently used. RESULTS: EEG and MEG findings following C fiber stimulation were similar to those following A delta fiber stimulation except for a longer latency. Finally, we reviewed the effect of rTMS on acute pain perception. rTMS alleviated acute pain induced by intracutaneous injection of capsaicin, which activated C fibers, but it enhanced acute pain induced by laser stimulation, which activated A delta fibers. CONCLUSIONS: One promising approach in the near future is to analyze the change of a frequency band. This method will probably be used for evaluation of continuous tonic pain such as cancer pain, which evoked response studies cannot evaluate. PMID- 15792884 TI - Subthreshold rTMS over pre-motor cortex has no effect on tics in patients with Gilles de la Tourette syndrome. AB - OBJECTIVE: A previous study showed no effect of 1Hz repetitive transcranial magnetic stimulation (rTMS) on tics in Gilles de la Tourette Syndrome (GTS). We modified the rTMS protocol in order to investigate some of the possible methodological reasons for the negative outcome in that study. METHODS: In a single blinded placebo-controlled cross-over study in five GTS patients without obsessive compulsive disorder we probed whether longer trains (1800 stimuli) of 1 Hz pre-motor cortex rTMS at 80% of active motor threshold and application to both hemispheres can improve tics in GTS. This was measured with the Yale Global Tic severity rating scale, the MOVES self-rating scale and video analysis. RESULTS: We found no significant effect of either left pre-motor cortex stimulation alone, or left pre-motor followed by right pre-motor cortex stimulation. CONCLUSIONS: These results suggest that the rTMS protocol used in this study is not useful for the treatment of tics in GTS. SIGNIFICANCE: rTMS protocols need to be modified substantially in order to explore their potential for the treatment of tics in GTS. PMID- 15792885 TI - Paradoxical modulation of tendon tap reflex during voluntary contraction in Parkinson's disease. AB - OBJECTIVE: Inadequate supraspinal modulation of spinal motor control mechanisms such as alpha-gamma coactivation is supposed to cause difficulty in maintaining proper voluntary contraction in Parkinson's disease (PD). METHODS: Subjects were 42 patients with PD and 20 normal volunteers. Soleus H-reflex and tendon tap reflex (T-reflex) were recorded. The maximal reflexes (H(max) and T(max)) at rest were recorded first. Next, the stimulus intensities were fixed to obtain a reflex size of around 25% of M(max) at rest for both H- and T-reflexes, and the reflexes were recorded at rest, during tonic plantarflexion (TPF), and at the onset of plantarflexion. RESULTS: H(max) at rest was 55% and T(max) 30% in normal subjects, while they were 36 and 31%, respectively, in PD. The size ratio of T(max) and H(max) at rest in PD was larger than normal. In PD, the size of H reflex increased with TPF as in normal subjects, but T-reflex decreased. These changes in T-reflex were correlated with the grade of rigidity, bradykinesia, and time for 10 m gait. H-reflex had no such correlations. CONCLUSIONS: T-reflex was abnormally modulated in PD especially during tonic contraction. SIGNIFICANCE: Inappropriate supraspinal modulation of the spinal reflex pathways disturbs motor performance in PD. PMID- 15792886 TI - Transcranial magnetic stimulation of deep brain regions: evidence for efficacy of the H-coil. AB - OBJECTIVE: Standard coils used in research and the clinic for noninvasive magnetic stimulation of the human brain are not capable of stimulating deep brain regions directly. As the fields induced by these coils decrease rapidly as a function of depth, only very high intensities would allow functional stimulation of deep brain regions and such intensities would lead to undesirable side effects. We have designed a coil based on numerical simulations and phantom brain measurements that allows stimulation of deeper brain regions, termed the Hesed coil (H-coil). In the present study we tested the efficacy and some safety aspects of the H-coil on healthy volunteers. METHODS: The H-coil was compared to a regular figure-8 coil in 6 healthy volunteers by measuring thresholds for activation of the abductor pollicis brevis (APB) representation in the motor cortex as a function of distance from each of the coils. RESULTS: The rate of decrease in the coil intensity as a function of distance is markedly slower for the H-coil. The motor cortex could be activated by the H-coil at a distance of 5.5 cm compared to 2 cm with the figure-8 coil. CONCLUSIONS: The present study indicate that the H-coil is likely to have the ability of deep brain stimulation and without the need of increasing the intensity to extreme levels that would cause a much greater stimulation in cortical regions. SIGNIFICANCE: The ability of non-invasive deep brain stimulation potentially opens a wide range of both research and therapeutic applications. PMID- 15792887 TI - The speed of sit-to-stand can be modulated in Parkinson's disease. AB - OBJECTIVE: To evaluate whether Parkinsonian patients could modify the speed of a sit-to-stand (STS) task to the same extent as that of healthy subjects. METHODS: Twenty Parkinsonian patients and 20 control subjects were instructed to stand up at a natural and fast speed. Kinematic data and kinetic data were recorded. RESULTS: Parkinsonian patients were significantly slower than healthy individuals during STS at a natural speed. When required to perform STS task at a fast speed, these patients could increase both peak horizontal and vertical velocities of the task, by significantly increasing hip and ankle dorsiflexion torques and the rate of torque production, just as the control subjects did. In fact, no difference was found for the percentage changes in both peak velocities and movement time between the two groups, though similar between-group differences during STS at a natural speed still existed at a fast speed. CONCLUSIONS: Parkinsonian patients had problems in generating adequate lower limb joint torques and in the rate of torque production when performing STS at a natural speed. However, these patients were capable of increasing the speed of their STS with the same percentage changes as those of healthy subjects. SIGNIFICANCE: The capability of Parkinsonian patients for increasing movement speed gives new insights to rehabilitation strategy. PMID- 15792888 TI - Lateralization of pupillary light reflex parameters. AB - OBJECTIVE: The aim of this study was to determine differing reactions of the left and right eyes with regard to pupillary light reflex (PLR) parameters. METHODS: All together 90 healthy subjects were included. In the first test series, 34 subjects were investigated on both eyes (left eye was tested first, three tests per day and one reliability test). In the second test series, 32 subjects were studied while changing the beginning side. In the last test series, 29 subjects were investigated 12 times each within 1h (beginning side changed, without spoken advice). Infrared pupillometry was used to study pupil diameter, latency time, relative amplitude, contraction/dilation velocity, and pupil redilation time. RESULTS: The study demonstrated significant differences of PLR parameters between both eyes. In contrast to the pupil diameter of the left eye the parasympathetically-dominated right eye was not influenced by vocal instructions or by changing the beginning side. CONCLUSIONS: PLR parameters might indicate functional lateralization of autonomic function in the central nervous system. High sensitivity of the procedure (arousal due to spoken advice, time of day) is advantageous for various psychophysiological investigations. SIGNIFICANCE: Differences between both eyes might point towards cortical lateralization of central autonomic function. PMID- 15792889 TI - Evaluation of commercially available electrodes and gels for recording of slow EEG potentials. AB - OBJECTIVE: To test the applicability of different types of commercially available electrodes and electrode gels or pastes for recording of slow EEG potentials. METHODS: Experiments were carried out on six types of reusable electrodes (silver, tin and gold cup electrodes, sintered silver-silver chloride (Ag|AgCl), platinum, stainless steel), six disposable Ag|AgCl electrode models, and nine gels or pastes. We studied the parameters, which are critical in slow-potential recording, such as polarization, initial and long-term stability and low frequency noise. RESULTS: The best results were obtained with the reusable sintered Ag|AgCl electrodes. The six disposable Ag|AgCl electrode models also proved to have appropriate electrical properties. Other types of reusable electrodes suffered from diverse degrees of polarization, baseline drift, low frequency noise, high resistance, and changes in properties due to wear and tear. Seven out of nine gels or pastes contained a significant amount of chloride, which is a prerequisite for DC stability of Ag|AgCl electrodes, whereas the absolute concentration of chloride had little effect. CONCLUSIONS: Direct current (DC) coupled recording of EEG is critically dependent on the choice of electrode and gel. SIGNIFICANCE: Our results provide rigorous criteria for choosing DC stable electrodes and gels for DC-coupled or long time-constant AC-coupled recordings of slow EEG potentials. PMID- 15792890 TI - Modulations of the visual N1 component of event-related potentials by central and peripheral cueing. AB - OBJECTIVE: The aim of this study is to investigate the effects of the central and peripheral cueing on N1 component of the event-related potentials (ERPs) and the time course of these effects. METHODS: ERPs were recorded while participants performed a discrimination task on the height of target bars, which were presented after informative-central, informative-peripheral or uninformative peripheral cues with stimulus onset asynchronies (SOAs) of 100, 300, 500 or 700 ms. RESULTS: Peripheral cues (informative and uninformative) elicited stronger effects of cue validity on N1 300 ms after cue onset, whereas central cues led to a more sustained validity effect on N1, with later stronger effects, at 500 ms SOA. CONCLUSIONS: The present data showed that central and peripheral cues affected to the level of processing reflected by the N1 component, but there were differences in the time course of these effects. Attentional orienting in response to central cueing resulted in a sustained validity effect on N1, relative to the more transitory activation of the process reflected by the N1 validity effect in this peripheral cueing task. SIGNIFICANCE: This study provides a detailed within-subject analysis of the time course of the effects of central and peripheral cueing on N1. PMID- 15792891 TI - Laser-evoked potentials: normative values. AB - OBJECTIVE: Laser-evoked potentials (LEPs) currently represent the most reliable and widely agreed method of investigating the A delta-fibre pathways. Many studies dealt with the usefulness of LEPs in peripheral and central nervous system diseases. We aimed at gaining normative values for LEP data. METHODS: Using a CO2 laser stimulator we recorded LEPs after face, hand, and foot stimulation in 100 normal subjects. We measured the perceptive threshold, latency and amplitude of the main vertex components, and their side-to-side differences. We also studied the correlations between LEP data and age and body height, as well as gender differences. RESULTS: Laser perceptive threshold increased and LEP amplitude decreased from face to foot (P<0.0001). The latency of hand and foot LEPs correlated significantly with body height (P<0.0001). The amplitude, though not the latency, correlated with age (P<0.0001). LEP data did not significantly differ between genders (P>0.1). CONCLUSIONS: This study provides normative values for the main LEP data and their absolute and side-to-side limits, highlighting the physiological differences related to, body height, age, gender and stimulation site. SIGNIFICANCE: Our data may help to improve the clinical reliability of LEPs as a diagnostic tool. PMID- 15792892 TI - Origins of the somatic N20 and high-frequency oscillations evoked by trigeminal stimulation in the piglets. AB - OBJECTIVE: In humans, the somatic evoked potentials (SEPs) and magnetic fields (SEFs) elicited by peripheral nerve stimulation contain high-frequency oscillations (HFOs) around 600 Hz superimposed on the initial cortical response N20. Responses elicited by snout stimulation in the swine also contain similar HFOs during the rising phase of the porcine N20. This study examined the generators of the N20 and HFOs in the swine. METHODS: We recorded intracortical SEPs and multi-unit activities in the sulcal area of the primary somatosensory cortex (SI) simultaneously with SEFs. The laminar profiles of the potential and current-source-density (CSD) were analyzed. RESULTS: The CSD analysis revealed that the N20 was produced by two dipolar generators, both directed toward the cortical surface. After the arrival of the initial thalamocortical volley in layer IV, the sink of the first generator shifted toward shallower layers II-III with a velocity of 0.109+/-0.038 m/s (mean+/-SD). The sink of the second generator moved to layer V. The initial thalamocortical axonal component of the HFO was produced by repolarizing current with the sink in layer IV. The CSD laminar profile of the postsynaptic component was very similar to the profile of intracortical N20. The current sink within each cycle of HFO propagated upward with a velocity of 0.633+/-0.189 m/s, indicating backpropagation. CONCLUSIONS: We propose that the N20 is generated by two sets of excitatory neurons which also produce the HFOs. Although the loci of synaptic inputs are unknown, these neurons appear to fire initially in the soma and produce backpropagating spikes toward distal apical dendrites. SIGNIFICANCE: These conclusions relate the N20 to the HFO and provide a new explanation of how the current underlying the N20 is invariantly directed toward superficial layers across species. PMID- 15792893 TI - Contralateral and ipsilateral responses in primary somatosensory cortex following electrical median nerve stimulation--an fMRI study. AB - OBJECTIVE: Ten healthy adult subjects were examined using functional magnetic resonance imaging (fMRI) to investigate responses in the contralateral and ipsilateral primary somatosensory cortex (SI) following electrical stimulation of the median nerve. METHODS: The right and left median nerves were stimulated alternately at the wrist in the different sessions. First, the location of the response in contralateral SI was identified following median nerve stimulation, and then, a spherical search volume with a 10mm radius centered on the region of the contralateral response was determined. Whether or not fMRI activation occurred within this sphere following ipsilateral stimulation was examined using a 3T MR imager. RESULTS: A response in contralateral SI was observed in 8 of the 10 subjects in right and left hemisphere. Responses in ipsilateral SI were observed in 6 of 8 subjects in right hemisphere, and the region of the response tended to be posterior to the contralateral region. On the other hand, in left hemisphere, the ipsilateral responses were found in three. CONCLUSIONS: In the present study, not only contralateral SI but also ipsilateral SI was activated following median nerve. The location of the ipsilateral activation was significantly more posterior than the contralateral one in right hemisphere. SIGNIFICANCE: The region of activation in ipsilateral SI was located in the posterior portion of post central gyrus, corresponding to around BA2 and 5 in human. PMID- 15792894 TI - Proprioceptive event related potentials: gating and task effects. AB - OBJECTIVE: The integration of proprioception with vision, touch or audition is considered basic to the developmental formation of perceptions, conceptual objects and the creation of cognitive schemes. Thus, mapping of proprioceptive information processing is important in cognitive research. A stimulus of a brisk change of weight on a hand held load elicit a proprioceptive evoked potential (PEP). Here this is used to examine early and late information processing related to weight discrimination by event related potentials (ERP). METHODS: A gating paradigm having 1s between the proprioceptive stimuli of 100 g weight increase was recorded in 12 runs of 40 pairs and an 1:4 oddball task of discriminating between 40 and 100 g weight increase was both recorded in 24 healthy men. The subjects were stratified in 3 groups according to their discrimination errors. RESULTS: The proprioceptive event related potential (PERP) consisted of a contralateral parietal P60, frontal N70, midline P100, initial contralateral later widespread N160, vertex P200, parietal N290 and target related widespread P360 and posterior N500. The target related components were augmented in the best performers, while the bad performers had delayed P60 and attenuated N70. The amplitudes of N160, P200 and N290 were unrelated to performance. Gating was seen as attenuation of P100, N160 and P200 amplitude. CONCLUSIONS: The proprioceptive stimulus feature processing seem to be accomplished in the first 100 ms, while later components are modified by context as expected from previous findings in the somatosensory modality. SIGNIFICANCE: The PERP could be a useful research tool in the investigation of bodily information processing in neuropsychiatric disorders. PMID- 15792895 TI - Gamma rhythms are not integral to EEG spindle phenomena. AB - OBJECTIVE: Gamma rhythms (30-100 Hz) have been shown to be associated with spindling activity induced by picrotoxin. To determine if gamma power is unique to picrotoxin spindles or is an integral part of physiological and pathological spindling activity we analysed and compared the strength and brain distribution of gamma EEG power during 4 spindling activities in the rat. METHODS: The electroencephalogram (EEG) was recorded from rats with chronically implanted electrodes during natural sleep, barbiturate anaesthesia, during naturally occurring absence epilepsy spike and wave discharges and following the systemic application of picrotoxin. Spectral analysis was applied off-line to compare the strength and brain distribution of gamma EEG power during the 4 spindling activities. RESULTS: Each spindle type contained significantly different levels of gamma power. Gamma power was significantly increased over background levels during picrotoxin spindles, slightly increased during absence spindles, slightly decreased during sleep spindles and significantly suppressed during barbiturate spindles CONCLUSIONS: Changes in the power of gamma frequencies during spindle types suggest that gamma frequencies are neither the cause of nor an integral part of a spindle. They appear to be correlated with levels of consciousness and may contribute to the process of epileptogenesis. SIGNIFICANCE: The findings are consistent with high frequency EEG activity being related to seizure-tendency. PMID- 15792896 TI - Progressive aging does not alter the interaction between autonomic cardiac activity and delta EEG power. AB - OBJECTIVE: We tested the hypothesis that the reductions of the changes in the respective influence of the cardiac sympathetic and vagal activity control and delta EEG activity with aging alter the interactions between the heart rate variability (HRV) and the delta sleep EEG power band. METHODS: A polysomnography was performed on 16 healthy young men and 19 healthy middle-aged men across the first 3 NREM-REM cycles. Spectral analysis was applied to electrocardiogram and electroencephalogram recordings. High Frequency (HF(nu)) of HRV as well as the maximum of cross-spectrum, coherency, gain and phase shifts between HF(nu) and delta sleep EEG power band were compared between both groups. RESULTS: Young men experienced more deep sleep than middle-aged men (P<0.001). In middle-aged subjects, HF(nu) was lower than the HF(nu) of their younger counterparts (P<0.001), but they showed similar increases during NREM sleep and similar decreases during REM sleep as the young subjects. Cross-spectrum values, coherency, gain and phase shifts between HF(nu) and delta were identical between the two groups. Modifications in HF(nu) show parallel changes and precede changes in delta EEG band by a similar leads of 11+/-6min in young men and 9+/-7 min in middle-aged men (P=0.23). CONCLUSIONS: Reduced changes in the respective influence of the cardiac sympathetic and vagal activity and delta EEG activity with progressive aging do not alter the relationship and phase difference between changes in the relative predominant cardiac vagal activity and delta power in middle-aged men. SIGNIFICANCE: Interaction between the cardiac sympathetic and vagal activity with delta EEG activity is maintained in middle-aged men. PMID- 15792897 TI - Effects of eyelid closure, blinks, and eye movements on the electroencephalogram. AB - OBJECTIVE: To characterize the effects of the eyeball and eyelid positions during eyeblinks on electroencephalographic (EEG) potentials. METHODS: Movements of the upper eyelids and eyes were measured in two healthy subjects using the magnetic search coil technique during horizontal and vertical eye rotations, eyeblinks, and lid closure. Corresponding signal changes were recorded simultaneously on the electroencephalogram (EEG). RESULTS: Spontaneous blinks produced small eye movements directed down and inward, whereas slow or forced blinks were associated with delayed upward eye rotations (i.e. Bell's phenomenon); both types of blinks caused positive EEG potentials with bifrontal distribution maximum at Fp1 and Fp2. CONCLUSIONS: In prior reports, these positive EEG artifacts have been attributed to upward eyeball rotation during blinks-Bell's phenomenon. By contrast, our findings indicate that movements of the eyelid contribute to a greater extent to these EEG potentials than do upward eyeball rotations. SIGNIFICANCE: Care is required in attributing EEG artifacts to movements of either eyeball or eyelid, since our findings suggest that they both contribute to these potentials. PMID- 15792898 TI - Anterior limbic alpha-like activity: a low resolution electromagnetic tomography study with lorazepam challenge. AB - OBJECTIVE: To verify findings of an independently regulated anterior limbic alpha band source. METHODS: In a randomised cross-over study, the spontaneous EEG was recorded in nine healthy subjects after i.v. lorazepam or placebo. Intracerebral current densities within classical frequency bands were estimated with low resolution electromagnetic tomography [LORETA] and compared between groups with t statistical parametric mapping [SPM[t]]. A region-of-interest [ROI] based method was used to compare frontal and occipital alpha band activity changes. RESULTS: Irrespective of treatment group, local maxima of alpha band power were localised both in the occipital lobe, Brodman area [BA] 18, and in the anterior cingulate cortex [ACC], BA 32. Statistical parametric mapping showed reduced parieto occipital, but unaltered frontal alpha band power after lorazepam. This result was confirmed by ROI-based comparison of BA 18 and BA 32. CONCLUSIONS: There was an anterior limbic maximum of alpha band activity which, unlike occipital alpha, was not suppressed by lorazepam. SIGNIFICANCE: The well-known anterior alpha band components may originate from a narrowly circumscribed source, located in the ACC. Frontal and occipital alpha band activities appear to be independently regulated. PMID- 15792899 TI - FVEPs in Creutzfeldt-Jacob disease: waveforms and interaction with the periodic EEG pattern assessed by single sweep analysis. AB - OBJECTIVE: To characterise flash visual evoked potentials (FVEPs) in 20 patients with Creutzfeldt-Jacob disease (CJD), and assess the relationships between spontaneous EEG patterns and the responses to individual stimuli. METHODS: We analysed the shape and time course of periodic sharp wave complexes (PSWCs) and responses to 1 Hz flashes. In nine patients, we applied an algorithm based on an autoregressive model with exogenous input (ARX) to estimate responses to individual random flashes and their interaction with PSWCs. RESULTS: The FVEPs included P1 and N1 components in all patients, and the P2 peak in 18. Eight patients showed giant FVEPs (N1-P2>60 V), all of whom had an MM polymorphism in codon 129 of the prion protein gene; in seven cases, the presence of giant FVEPs correlated with a prominent and almost continuous periodic EEG pattern. Giant N1 P2 abnormally spread on the anterior scalp regions, and had a different waveform distribution from that of the PSWCs. In five patients with a normal or slightly enlarged average N1-P2 amplitude, single sweep (ARX) analysis revealed a period of relative refractoriness following individual PSWCs. In four patients with 'giant' FVEPs, the individual responses occurred regardless of the interval between the stimulus and previous PSWC, but their amplitude had an inverse relationship with the interval length. CONCLUSIONS: Giant responses to flash stimuli are a common finding in CJD patients (40% of our cases). Single sweep ARX analysis showed that PSWCs were followed by a period of partial refractoriness, which prevented most of the individual responses to flashes, but not giant FVEPs. The association between prominent spontaneous paroxysms and giant FVEPs suggests that both are due to a common hyperexcitable change favouring neuronal synchronisation. SIGNIFICANCE: Our data contribute to clarifying the debated problem of the occurrence of giant FVEPs in CJD and their relationships with the spontaneous periodic EEG pattern. PMID- 15792900 TI - Comparison of clonidine to sleep deprivation in the potential to induce spike or sharp-wave activity. AB - OBJECTIVE: This study aimed to investigate previously observed side effects, i.e. increased epileptic activity during clonidine medication. The safety and effectiveness of clonidine as spike inducing agent compared to sleep deprivation were tested. METHODS: Patients suffering from drug-resistant localization related epilepsy took part in 3 magnetoencephalography (MEG) sessions. One session was recorded without any activating measures. The other two sessions were either performed after sleep deprivation or after medication with clonidine. Target parameter was the number of spikes or sharp-waves during a 30 min recording period. RESULTS: About 67% of the patients showed increased spike activity after clonidine, whereas sleep deprivation increased the number of spikes in 33%, and 29% of the patients did not show any activation at all. Clonidine was most effective in temporal lobe epilepsy, when the focus was located in the right hemisphere, and when clonidine serum concentrations were ranging between 0.6 and 1.0 ng/ml. CONCLUSIONS: This study showed for the first time that clonidine can be considered an effective spike or sharp-wave inducing drug that is superior to the potency of sleep deprivation. SIGNIFICANCE: The administration of clonidine increases the probability of recording ictal and interictal epileptic activity during limited acquisition time. PMID- 15792901 TI - Independent component analysis of generalized spike-and-wave discharges: primary versus secondary bilateral synchrony. AB - OBJECTIVE: To differentiate between primary and secondary bilateral synchrony (PBS and SBS, respectively) in generalized spike-and-wave discharges (GSWD), we applied independent component analysis (ICA) to GSWD, and analyzed the characteristic patterns of independent components. METHODS: EEGs from 19 patients with GSWD (9 PBS patients, 10 SBS patients) were studied. Thirty GSWD epochs were selected and concatenated to construct an EEG data matrix that was subjected to ICA. Selected independent components were localized by mapping them on a spherical model of the head by means of brain electrical source analysis (BESA) to define dipole sources. RESULTS: Epileptic components of GSWD were clearly separated by the ICA algorithm in all patients. Between one and three components per patient were responsible for GSWD. In PBS patients, 70.6% of the independent components had dipole sources within the dorsolateral frontal region, and 56.5% of the independent components in SBS patients were within the medial frontal region (P=0.012). The orientation of all of the independent components in PBS patients was radial, whereas 47.8% of the independent components in SBS patients were tangential; this difference was statistically significant (P=0.001). CONCLUSIONS: Spatiotemporal decomposition of GSWD by using ICA might be helpful for differentiating SBS from PBS. SIGNIFICANCE: The localization of the dipole sources of the independent components may provide insight into the pathophysiological origins of GSWD. PMID- 15792902 TI - Assessing cortical functional connectivity by linear inverse estimation and directed transfer function: simulations and application to real data. AB - OBJECTIVE: To test a technique called Directed Transfer Function (DTF) for the estimation of human cortical connectivity, by means of simulation study and human study, using high resolution EEG recordings related to finger movements. METHODS: The method of the Directed Transfer Function (DTF) is a frequency-domain approach, based on a multivariate autoregressive modeling of time series and on the concept of Granger causality. Since the spreading of the potential from the cortex to the sensors makes it difficult to infer the relation between the spatial patterns on the sensor space and those on the cortical sites, we propose the use of the DTF method on cortical signals estimated from high resolution EEG recordings, which exhibit a higher spatial resolution than conventional cerebral electromagnetic measures. The simulation study was followed by an analysis of variance (ANOVA) of the results obtained for different levels of Signal to Noise Ratio (SNR) and temporal length, as they have been systematically imposed on simulated signals. The whole methodology was then applied to high resolution EEG data recorded during a visually paced finger movement. RESULTS: The statistical analysis performed returns that during simulations, DTF is able to estimate correctly the imposed connectivity patterns under reasonable operative conditions, i.e. when data exhibit a SNR of at least 3 and a length of at least 75 s of non-consecutive recordings at 64 Hz of sampling rate, equivalent, more generally, to 4800 data samples. CONCLUSIONS: Functional connectivity patterns of cortical activity can be effectively estimated under general conditions met in any practical EEG recordings, by combining high resolution EEG techniques, linear inverse estimation and the DTF method. SIGNIFICANCE: The estimation of cortical connectivity can be performed not only with hemodynamic measurements, by using functional MRI recordings, but also with modern EEG recordings treated with advanced computational techniques. PMID- 15792903 TI - Detection of EEG background abnormalities in epilepsy by a new spectral index. AB - OBJECTIVE: In epileptic disorders, EEG background activity is disorganized in or near the epileptogenic focus and spectral EEG analysis (SEA) can provide useful information about the focus. We tried to develop a new spectral index from basic spectral parameters to detect the epileptic abnormalities at EEG background activity. METHODS: A new spectral EEG index, epileptic abnormality index (EAI), was constructed from frequency band power and power asymmetry parameters. Within the index, parameters were weighted due to both conventional EEG knowledge and their power in discrimination healthy subjects from patients. EEG background activity from 99 epileptic patients and 146 healthy subjects was examined both by EAI and by a conventional SEA method, by using z-scoring statistic. Each test results were compared with visual EEG interpretation of subjects. RESULTS: In patient groups, EAI was most successful in lateralization of epileptic abnormalities. It was also helpful in discrimination of epileptic patients from normals in the case where visual EEG interpretation was 'normal'. CONCLUSIONS: EAI depends on basic spectral parameters and it combines statistical methods and clinical knowledge about EEG. It increases the analysis capacity of SEA in evaluation of EEG background activity. SIGNIFICANCE: EAI is a new and useful approach in detection of EEG background abnormalities in epilepsy and its logical base can also be used in the detection of brain electrical activity abnormalities other than epileptic disorders. PMID- 15792904 TI - Evaluation of the complexity of motor unit potentials in anal sphincter electromyography. AB - OBJECTIVE: Motor unit potential (MUP) morphology can be quantified using parameters describing electrophysiological size and shape (complexity). Traditionally, MUP complexity has been estimated using parameters number of phases and turns. Recently, 'irregularity coefficient' (IR), measuring the length of the MUP curve normalized with its amplitude, has been introduced. The aim of this study was to evaluate IR in the external anal sphincter muscle. METHODS: Sensitivity was examined in 61 patients with chronic cauda equina lesions, and specificity in 75 controls using a standard concentric EMG needle and EMG system with multi-MUP analysis. RESULTS: When evaluated separately the sensitivity of IR was 16% lower, and specificity 5% higher compared to number of turns, with both differences decreasing to only 2 and 1%, respectively, when each of these parameters was added to MUP area and duration. CONCLUSIONS: Our present results suggest that IR provides a similar diagnostic usefulness, but a more appropriate description of MUP complexity compared to traditional MUP parameters. SIGNIFICANCE: IR seems to be suitable to complement one of MUP parameters measuring electrophysiological MUP size in the future quantitative EMG. PMID- 15792905 TI - The SIIR index--a non-linear combination of waveform size and irregularity parameters for classification of motor unit potentials. AB - OBJECTIVE: In clinical electromyography, motor unit potential (MUP) size is used to indicate the myo- or neurogenic origin of the pathological process, whereas the irregularity of MUP shape is considered not useful in this respect. The aim of the present study was to find a method to improve classification of MUPs using a combined analysis of the size and irregularity of the potential. METHODS: The records with at least 20% of irregular potentials from 14 healthy subjects and 34 patients were analysed. Principal Component Analysis (PCA) was applied in the analysis of data to find a linear function combining the size and the irregularity of a MUP. RESULTS: The linear combination of size and irregularity measures, i.e. size index (SI) and irregularity coefficient (IR) obtained using Principal Component Analysis (PCA) did not increase the discriminating ability in comparison to SI. We have defined a non-linear function of the SI and IR called SIIR, which modifies SI values for irregular potentials only. Its discriminating ability is higher than that of SI. CONCLUSIONS: By combined assessment of the size of a MUP and its irregularity, the differentiation between neurogenic and myopathic potentials could be considerably improved. SIGNIFICANCE: The proposed coefficient SIIR seems to be more discriminating than others presently used. When applied in clinical studies it may be useful in the evaluation of confusing, atypical motor unit potentials. PMID- 15792906 TI - Electrophysiological features of patients with POEMS syndrome. AB - OBJECTIVE: Polyneuropathy, organomegaly, endocrinopathy, M protein, and skin changes (POEMS) syndrome is a rare cause of polyneuropathy. Although the polyneuropathy component is essential for the diagnosis of the disease, the pattern of associated electrodiagnostic abnormalities has not been characterized in detail. The purpose of this study was to elucidate the features of nerve conduction abnormalities in POEMS syndrome. METHODS: We reviewed the medical records and nerve conduction studies (NCS) of 12 consecutive patients with POEMS. RESULTS: A total of 68 motor and 46 sensory nerves were examined. Compound muscle action potentials (CMAPs) and sensory nerve action potentials (SNAPs) were not elicited in lower limbs more frequently compared with in upper limbs (P<0.05, in both motor and sensory nerves), and the CMAP amplitude was more attenuated in lower limbs than in upper limbs (P<0.05). Abnormal conduction slowing was frequently observed in motor (95%) and sensory (76%) nerves. Distal motor latencies were abnormally prolonged in 75% of the motor nerves, and terminal latency indices were significantly higher in patients than in normal controls (P<0.05). CONCLUSIONS: NCS in POEMS syndrome showed characteristic patterns, in which conduction abnormalities were more frequently and severely affected in the lower limbs, and more predominantly in the intermediate nerve segments than in the distal portions. SIGNIFICANCE: The recognition of these characteristic patterns may be helpful for the early diagnosis of polyneuropathy in POEMS syndrome. PMID- 15792907 TI - A reappraisal of various methods for measuring motor nerve refractory period in humans. AB - OBJECTIVE: To compare various techniques of stimulation and methods of analysis to estimate absolute (ARP) and relative (RRP) refractory periods in motor nerve trunks of humans. METHODS: Double collision (DC) technique and two types of paired pulse (PP) technique, with test stimulation of supramaximal (PP(supra)) or submaximal (PP(sub)) intensity, were applied to 32 healthy subjects. The ulnar nerve was stimulated either at a single site (wrist) for the PP techniques or at two sites (wrist and elbow) for the DC technique, with various distal interstimuli intervals (ISIs). The elicited compound muscle action potentials (CMAPs) were recorded from the abductor digitorum minimi muscle. The DC technique provided estimates of minimal and maximal ARPs, whereas maximal RRP values were obtained with the PP techniques. Data were analyzed using three methods: a visual reading of the raw ISI-CMAP curves and two computer-aided analyses of the regression curve fitting the ISI-CMAP plots. Pain induced by each technique was assessed on a 0-10 visual analogue scale. A test-retest study was performed with the PP techniques in 12 subjects. RESULTS: RP estimates varied with both the stimulation technique and the analysis method. The DC technique was more painful than the PP techniques, but provided shorter and more accurate ARP values, whereas the PP(sub) technique provided longer, but valid RRP values. Computer aided methods of data analysis gave the lowest coefficients of test-retest variation. CONCLUSIONS: Compared to the PP techniques, the DC technique allowed the evaluation of the whole distribution of ARP estimates, not distorted by muscle fiber RPs. For RRP estimation, the PP(sub) technique can be preferred to the PP(supra) technique. Finally, computer-aided methods are preferable to analyze ISI-CMAP curves. SIGNIFICANCE: The distribution of RP estimates can be easily and reliably assessed in whole motor nerve trunks of humans, providing valuable information to assess peripheral nerve excitability. PMID- 15792908 TI - Age and gender effects in EEG coherence: II. Boys with attention deficit/hyperactivity disorder. AB - OBJECTIVE: This study investigated intra-hemispheric and inter-hemispheric EEG coherences as a function of age in boys with different subtypes of Attention Deficit/Hyperactivity Disorder (AD/HD), in comparison with a control group of normal boys. METHODS: Three groups of 40 children (AD/HD combined type, AD/HD inattentive type, and normal controls) participated. Each group contained 8 males in each 1 year band from 8 to 12 years. EEG was recorded from 21 sites during an eyes-closed resting condition. Wave-shape coherence was calculated for 8 intra hemispheric electrode pairs (4 in each hemisphere), and 8 inter-hemispheric electrode pairs, within each of the delta, theta, alpha and beta bands. RESULTS: Developmental effects in intra-hemispheric coherences at shorter and longer inter electrode distances generally supported Thatcher's two-compartment model. Control boys showed evidence of development in longer-range inter-hemispheric coherences which was not apparent in AD/HD boys. Boys with AD/HD of the combined type showed qualitatively different anomalies than boys with AD/HD of the inattentive type. CONCLUSIONS: EEG coherences in normal boys of this age range develop systematically with age in a non-linear fashion. Boys with AD/HD do not show this development. They display coherence anomalies which differ in nature between DSM IV subtypes, suggesting differences which are not relatable to simple symptom severity. SIGNIFICANCE: The data reported here indicate differences in the development of EEG coherences in boys with AD/HD, and point to differences between the AD/HD subtypes which may help understanding of the disorder. PMID- 15792909 TI - Auditory mismatch negativity is a good predictor of awakening in comatose patients: a fast and reliable procedure. PMID- 15792910 TI - Acute repetitive pilomotor seizures. PMID- 15792912 TI - Importance of biologic follow-ons: experience with EPO. AB - The importance of recombinant human erythropoietin (epoetin) therapy has been clearly demonstrated in patients with anemia due to chronic kidney disease. The use of biopharmaceuticals to replace endogenous proteins, which may be inadequately low, carries the risk of stimulating the immune system to develop autoantibodies. Although these proteins are designed to closely mimic the endogenous proteins, they may have potential immunogenic properties. Erythropoietin produced by recombinant DNA technology is the most successful and efficacious agent for treating anemia. It was initially used in treating anemia in chronic kidney disease patients. Pure red cell aplasia (PRCA) ensuing from production of neutralizing anti-erythropoietin antibodies occurred very rarely with epoetin treatment. This agent was initially administered intravenously, but the mode of administration was progressively altered to subcutaneous without apparent increase in immune reaction. However, between 1998 and 2001, a sharp increase in the number of PRCA cases was seen. PRCA had been a very rare complication until this time. All of these patients had high affinity neutralizing anti-erythropoietin antibodies. This observation was made primarily in cases where one brand of epoetin, Eprex, was administered subcutaneously to patients with chronic kidney disease treated outside the United States, although a small number of cases among chronic kidney disease patients treated solely with epoetin beta were also identified. The marked increase in the number of Eprex cases was attributed to a change in the stabilizers, storage, and route of administration of Eprex to patients with chronic kidney disease. Since then changes have been made to the route of administration, storage, and handling of Eprex, and more recently to the rubber stoppers used in the prefilled syringes. Eprex was administered intravenously to chronic kidney disease patients and, with improved storage and handling, there was a subsequent dramatic reduction in the number of cases. More recently, the rubber stoppers have been replaced by Teflon coated ones to prevent interactions with stabilizers and release of chemicals that have adjuvant properties. However, this concern is still relevant in the new generation of epoetin agents and generic formulations of epoetins. Epoetin treatment for anemia requires regular follow-up of hemoglobin levels but also of reticulocyte counts in chronic kidney disease patients. PMID- 15792913 TI - Recombinant human erythropoietin in cancer-related anemia: an evidence-based review. AB - Anemia is a common complication of cancer or anticancer therapy, with a significant negative impact on the functional status of patients and their quality of life (QOL). Recombinant human erythropoietin (EPO) was developed in the 1980's and was initially developed for the treatment of anemia associated with chronic renal failure. Subsequently, randomized, placebo-controlled clinical trials in the oncology setting were performed in the early 1990's. These studies demonstrated in cancer patients that EPO could increase the levels of hemoglobin (Hb), decrease the need for red blood cell transfusions, and also suggested that these outcomes were associated with improved QOL metrics as reported by the patients themselves. Based on the results of these studies, EPO was granted regulatory approval to be used for the treatment of anemia in patients with non myeloid malignancies where anemia was due to the effect of concomitantly administered chemotherapy. To further expand the findings of the registration studies, and to develop more complete QOL data, three similarly designed open label community-based studies were performed, enrolling a total of approximately 7000 patients. These studies consistently demonstrated in the supportive care of cancer patients receiving chemotherapy that the use of EPO could induce increases in the levels of Hb and that these correlated with patient-reported improvements in QOL metrics. The correlation between an improvement in Hb and QOL has also been confirmed in a larger randomized, placebo-controlled trial. Careful analyses of data from this study also helped support earlier findings that EPO could be effective for patients with varying degrees of anemia, and that increasing and maintaining Hb levels close to the physiologically normal levels resulted in the optimal improvements in QOL. While further investigations of EPO as a mechanism to improve the antineoplastic efficacy of chemoradiotherapy have not yet been positive, the overall experience with this agent remains very favorable after extensive studies and long-term clinical use in oncology. PMID- 15792914 TI - Phase III clinical trials with darbepoetin: implications for clinicians. AB - Anemia occurring in patients with renal failure on dialysis has been shown to both increase transfusion requirements and decrease functional status and quality of life. As a result, treatment of these patients with an erythropoietic agent such as epoetin alfa or darbepoetin alfa has become established as an essential part of optimal patient care. Anemia resulting from cancer or chemotherapy is a common problem in oncology, and has been shown similarly to increase transfusion risk and decrease patient reported outcomes. For reasons that are not clear, but may include either frequently encountered slow response or nonresponse and higher doses and hence higher costs, treatment of anemia in oncology has not become a standard treatment for all patients. Erythropoietic agents (epoetin alfa and darbepoetin alfa) have been used to improve anemia-related fatigue and quality of life in cancer patients. Epoetin is administered 1 to 3 times per week whereas darbepoetin, with up to a 3-fold greater half-life, is given once every 1 or 2 weeks. Recent data demonstrate that darbepoetin can be administered as infrequently as every 3 weeks with comparable erythropoietic efficacy. Several studies have been carried out to determine the optimum schedule of dosing to obtain maximum patient benefit. Following treatment with darbepoetin, antibodies to darbepoetin were not detected, no unusual adverse event was seen with darbepoetin, and the mean increase in hemoglobin remained unchanged when the dosing interval was increased from 1 to 2 weeks. The safety and efficacy of darbepoetin was also determined in patients with solid tumors receiving chemotherapy; the lowest clinically effective dose was determined to be 3.0 and 5.0 microg/kg every 2 weeks with a 66 and 84% response, respectively. No additional benefit was seen with higher doses. A multicenter study evaluated the safety and efficacy of darbepoetin administered either weekly, every 3 weeks, or every 4 weeks in anemic patients with cancer who were not receiving chemotherapy or radiotherapy. The results indicated that with weekly dosing, 70% of patients showed an increase in hemoglobin. The dose of 4.5 microg/kg/week resulted in 100% hematopoietic response. In a randomized, active control, pilot trial, front loading darbepoetin followed by lower doses or less frequent doses also appears to be efficacious and may decrease the time to response. At the present time, darbepoetin improves anemia, reduces requirements for transfusion, and improves the quality of life for these patients. To compare efficacy of these erythropoietic agents it is important to analyze the data using intent to treat rather than analysis of patients who complete the study. This way a true evaluation of the change of hemoglobin can be assessed corresponding both to dose and an improvement in the QOL. PMID- 15792915 TI - Anemia and the elderly: clinical considerations. AB - Anemia is a common problem in the management of elderly patients. Anemia in the elderly is linked to an increase in morbidity and mortality, and serves as an independent variable associated with poor outcomes. In the elderly population, anemia is a risk factor for cardiovascular health and early death, contributes to fatigue, and negatively impacts on cognitive function, physical function, and the quality of life and serves as a marker of increased vulnerability. A greater understanding of pathophysiology of anemia in the setting of chronic disease has provided insights into the rationale for the potential clinical application of erythropoietic agents in the treatment of anemia. The impact of erythropoietic therapy, as it relates to specific afflictions affecting the older age patient, including diminished exercise tolerance, frailty, loss of functional capacity, immobility, depression, increased care needs and cardiovascular fitness, remains to be determined and is worthy of further investigation. PMID- 15792916 TI - The blue cross blue shield assessment technology review: summary of findings. AB - Epoetin (EPO; erythropoietin) treatment offers an attractive but costly alternative to red blood cell transfusion for managing anemia associated with cancer therapy. The goal of this paper is to review the findings of the 2001 Blue Cross/Blue Shield Association Technology Evaluation Center overview on EPO use in oncology, which served the foundation for the 2002 ASH/ASCO clinical guideline on this subject. The BC/BS review had two major aims: (1) to quantify the effects of EPO on the likelihood of transfusion and on quality of life in patients with cancer treatment-related anemia, and (2) to evaluate whether outcomes are superior when EPO treatment is initiated at higher hemoglobin thresholds. Two independent reviewers followed a prospective protocol for identifying relevant studies published between 1985 and 1998. Outcomes data were combined with the use of a random-effects meta-analysis model. Double-blind, randomized controlled trials that minimized patient exclusions were defined as higher quality for sensitivity analysis; randomized, but unblinded trials and trials with excessive exclusions were included in the meta-analysis, but were defined as lower quality. Twenty-two trials (n=1927) met inclusion criteria, and 12 (n=1390) could be combined for estimation of odds of transfusion. EPO decreased the percentage of patients transfused by 9-45% in adults with mean baseline hemoglobin concentrations of 10g/dL or less (7 trials; n=1080), by 7-47% in those with hemoglobin concentrations greater than 10g/dL, but less than 12g/dL (7 trials; n=431), and by 7-39% in those with hemoglobin concentrations of 12g/dL or higher (5 trials; n=308). In a sensitivity analysis, the combined odds ratio for transfusion in EPO-treated patients as compared with controls was 0.45 (95% confidence interval [CI]=0.33-0.62) in higher quality studies and 0.14 (95% CI=0.06-0.31) in lower quality studies. The number of treated patients needed to prevent one transfusion is 4.4 for all studies, 5.2 for higher quality studies, and 2.6 for lower quality studies. Only studies with mean baseline hemoglobin concentrations of 10g/dL or less reported statistically significant effects of EPO treatment on quality of life; quality-of-life data were insufficient for meta analysis. No studies addressed the effects of EPO on anemia-related symptoms. The review concluded that EPO reduced the odds of transfusion for cancer patients undergoing therapy. Evidence was insufficient to determine whether initiating EPO earlier spares more patients from transfusion or results in better quality of life than waiting until hemoglobin concentrations decline to nearly 10g/dL. An update of this review, which includes information on quality of life, transfusion requirment, survival and tumor response, for first and second-generation epoetin products is ongoing. PMID- 15792917 TI - The ASH/ASCO clinical guidelines on the use of erythropoietin. AB - The only combined effort between the American Society of Hematology (ASH) and the American Society of Clinical Oncology (ASCO) in practice guideline development was in studying the use of erythropoietin in chemotherapy-related anemia and in myelodysplasia. This process began with an application to the Agency for Health Care Policy and Research (AHCPR, now called the Agency for Healthcare Research and Quality, or AHRQ) jointly from ASH and ASCO. As part of a competitive review process, the topic was selected by the Agency for funding. Subsequently, as part of the Agency's Evidence-based Practice program, funding to conduct the review was awarded to the Evidence-based Practice Center of the Blue Cross/Blue Shield Technology Evaluation Center, which performed an extensive literature review using principals of evidence-based medicine and with input from representatives of ASH and ASCO. That evidence-based review was then shared with an ASH and ASCO guidelines committee funded by both organizations and made up of members from both organizations. The guideline was developed over a period of 21/2 years, culminating in simultaneous publication in both Blood and The Journal of Clinical Oncology in October 2002. This field is in flux and much of the guideline discussed potential future directions for research. Projected research topics include whether a hemoglobin level of 11g/dl as a clinical trigger point has clinical benefit; whether increasing hemoglobin levels greater than 12g/dL has clinical benefit; what role iron supplementation plays in erythropoietin treated patients; weekly versus more frequent dosing; and cost benefit analyses. Quality of life considerations are important, but the practice guideline committee felt that there was not enough data in this area to view it with the same importance as avoidance of transfusion or rises in hemoglobin value as therapeutic goals for patients being treated with erythropoietin. PMID- 15792918 TI - Evidence-based medicine: can it be applied to stimulation of erythropoiesis for patients with malignancy? AB - Health care decision-making is affected by the values of patients and providers, available resources, and information substantiating effectiveness (or efficacy) of a particular therapy. A number of factors contribute to our growing need for evidence-based decision-making. Our aging population generally requires greater medical attention in a system with limited resources devoted to health care. Technologic advances have produced an ever-expanding range of expensive treatment options. Patients, and their providers, expect early access to these emerging therapies. Patients also expect care of universally high quality, even as respected authorities suggest there exists a substantial gap between these expectations and the care actually delivered. Evidence based decision-making offers the opportunity to use medical evidence to reduce uncertainty regarding research information and improve the value of health care delivered. Evidence based medicine (EBM) is the 'conscientious, explicit and judicious use of current best evidence in making decisions about the care of individual patients.' It combines clinical judgment and experience, best available scientific evidence, and patient preferences to improve medical decision-making. Though often incorrectly maligned as cookbook medicine that dismisses any research findings that do not derive from randomized clinical trials, it provides clinicians, health care systems, payers and policymakers with tools to appropriately evaluate the research evidence that substantiates various therapies and integrate that evidence with clinical expertise and patient's values in medical decision-making. The erythropoietic stimulants epoetin and darbepoetin have shown efficacy in improving anemia of chronic renal failure and following chemotherapy for many patients. In some studies these agents have also improved health-related quality of life. Unfortunately, only 60-80% of patients treated with erythropoietic stimulants respond, and like many emerging therapies, they are quite expensive. Likewise, there is substantial variation in their usage, suggesting both inappropriate use and non-usage. Reimbursement coverage decisions for erythropoietic stimulants have been hampered by the lack of high-quality evidence in certain applications. Physicians are increasingly turning to evidence-based medicine resources (guidelines, systematic reviews) to inform their decisions regarding application of new therapies. Evidence-based medicine offers health care decision-makers the opportunity for quality improvement, efficient resource allocation and utilization, informed policy-making and reimbursement, and identification of future research priorities. Judicious use of erythropoietic stimulants guided by evidence-based decision-making should ensure treatment of patients who can be reasonably expected to benefit with appropriate dose regimens, while preserving valuable health care resources in those situations where patients are not expected to derive significant health care benefit. PMID- 15792919 TI - Effectiveness of erythropoietin in the treatment of patients with malignancies: methods and preliminary results of a Cochrane review. AB - Cancer and cancer therapy-associated anemia may have an impact on tumor response and overall survival. Additionally, anemia represents an important economic factor. Therefore, therapeutic alternatives such as erythropoietin (EPO) and red blood cell transfusions have to be evaluated systematically. The effectiveness of recombinant human EPO to prevent or alleviate anemia in patients with malignant disease was determined. Randomized controlled trials comparing prophylaxis or treatment of anemia with EPO plus red blood cell transfusion (RBCT) or RBCT only in patients with malignant disease undergoing antineoplastic therapy were included. The endpoints needed for RBCT were hematological response (hemoglobin increase of 2g/dL or hematocrit increase of 6%), tumor response, and overall survival. Medical databases (Cochrane Library, MEDLINE, EMBASE) and conference proceedings were searched (1985-2001). Full-text and abstract publications were included as well as unpublished data. Data extraction and quality assessment were done in duplicate. All authors were contacted to obtain missing data. Out of 33 eligible studies, 27 trials with 3,287 randomized patients were included. PMID- 15792920 TI - Erythropoietin use in oncology: a summary of the evidence and practice guidelines comparing efforts of the Cochrane Review group and Blue Cross/Blue Shield to set up the ASCO/ASH guidelines. AB - Clinical research evidence on outcomes of using epoetin (EPO) to treat or prevent anemia in oncology has recently been systematically synthesized to provide a scientific foundation for developing and implementing clinical practice guidelines. Two groups have distinguished themselves by their meticulous research methods, the Blue Cross and Blue Shield Association Technology Evaluation Center (BCBSA TEC) and the Cochrane Review Group (CRG), and have summarized existing research evidence on the role of EPO in anemia associated with cancer treatment. An ASH/ASCO (American Society of Hematology/American Society of Clinical Oncology) panel has used the BCBSA TEC review to develop practice guidelines on the use of EPO in patients with cancer. The ASH/ASCO guideline panel identified eight important clinical circumstances for which use of EPO in oncology might be considered and used the BCBSA TEC evidence review to formulate evidence-based guidelines that support use of EPO. Both BCBSA TEC and CRG found solid evidence exists to show that EPO improves hemoglobin levels and reduce the risk for transfusion. The ASH/ASCO panel concluded that best empirical evidence exists to support the use of EPO to correct anemia due to chemotherapy if Hgb95% of the cases, EPREX had been administered subcutaneous to persons with chronic kidney disease (CKD) and anemia for a mean of nine months prior to diagnosis of PRCA. For 80% of persons with the syndrome, reversal of antibody production and recovery of reticulocytes occurred with discontinuation of epoetin and treatment with immunosuppressive agents. Patients with anemia of CKD who developed neutralizing anti-erythropoietin antibodies and pure red cell aplasia during treatment with epoetin have been identified in a number of countries. In non-US countries, switching renal dialysis patients from subcutaneous to intravenous administration of epoetin alpha and improved handling of the drug appear to have been successful strategies for reducing the occurrence of this toxicity. The decrease in cases occurred coincident with these varied changes, although it is difficult to prove causality. PRCA is a rare, but important side effect of epoetin therapy. PMID- 15792925 TI - Angiotensin II-induced hypertension in mice caused an increase in insulin secretion. AB - OBJECTIVE AND DESIGN: Insulin action was determined in a mouse model of human hypertension via chronic angiotensin II administration followed by a glucose tolerance test. METHODS: Angiotensin II or saline was infused systemically into mice via osmotic pump for 2 or 4 weeks. In angiotensin II-treated mice versus saline controls we compared blood pressure, blood glucose, and serum insulin concentrations during an intravenous glucose tolerance test and assessed glucose transport and insulin signaling in muscle. RESULTS: Blood pressure increased at 2 and 4 weeks following angiotensin II treatment. Mice treated with angiotensin II for 4 weeks cleared a glucose bolus faster than mice treated with saline despite similar basal serum insulin concentrations. Upon glucose administration, the increase in serum insulin was greater in angiotensin II-treated mice, 38.8+/-6.5 pmol/l, compared to saline-treated mice, 21.8+/-2.9 pmol/l, but only at 4 weeks of angiotensin II treatment while no difference was observed at 2 weeks of angiotensin II administration. At 4 weeks of angiotensin II treatment, insulin signaling in the liver and in the skeletal muscle was not affected, since both the number of insulin receptors and phosphorylation of Akt were unchanged. Also at 4 weeks of angiotensin II treatment, ex vivo soleus muscle did not exhibit any change in basal and insulin-stimulated glucose uptake. CONCLUSIONS: This study suggests that long-term angiotensin II treatment for 4 weeks enhances glucose stimulated insulin secretion in mice. Angiotensin II-induced hyperinsulinemia may play a role in the development of insulin resistance in patients with hypertension. PMID- 15792926 TI - Effects of early perturbation of the renin-angiotensin system on cardiovascular remodeling in spontaneously hypertensive rats. AB - RATIONALE: The goal of this study was to analyze cardiovascular (CV) remodeling in early, short-term CAP treated SHR and their offspring. METHODS: We treated SHR with Captopril (CAP, 100 mg/kg) from in utero to 1 month of age (OCAP). Some of these rats were mated at 3-4 months of age and we used their offspring (2nd G). Controls were untreated SHR, normotensive Wistar Kyoto rats (WKY) and SHR maintained on CAP (SCAP). At 12-14 months of age, rats were cannulated for mean arterial blood pressure (MAP) measurements. An image analysis system was used to quantitate changes in cardiac and vascular (wall-to-lumen ratios, w/l) morphology and fibrosis. RESULTS: Early, short-term CAP treatment prevented the full expression of hypertension in treated rats and their offspring. MAPs were: SHR (180+/-2.2 mm Hg); WKY 125+/-3 mm Hg); SCAP 112+/-2.5mm Hg; OCAP 138+/-2.3 mm Hg; and 2nd G (145+/-2.0 mm Hg). There were significant decreases in heart weight/body weight ratios, large and small vessel morphology, and interstitial and perivascular fibrosis in CAP-treated animals and their offspring in comparison to untreated SHR. CONCLUSIONS: The CV protective properties of early, short-term CAP treatment were not solely due to a reduction in MAP. Although MAP was higher in OCAP and 2nd G, CV structure resembled that found in WKY and SCAP. The effects of our early treatment appear to be due to chronic blockade of the renin-angiotensin system and its effects on growth of CV tissues and the development of fibrosis. PMID- 15792927 TI - Expression, localisation and function of ACE and chymase in normal and atherosclerotic human coronary arteries. AB - The expression, localisation and function of enzymes responsible for the local formation of angiotensin II in atherosclerotic and non-atherosclerotic human coronary arteries were studied. Human epicardial coronary arteries expressed mRNA for both ACE and chymase. Immunohistochemical studies revealed that ACE was localised to the vascular endothelium, and to a lesser extent the medial smooth muscle cells, in both large and small arteries. Chymase was detected in both types of vessel but was shown to be associated with mast cells. The contractions to angiotensin I in large arteries were inhibited only by a combination of quinaprilat and soyabean trypsin inhibitor. In the intramyocardial arteries the response to angiotensin I was markedly inhibited in the presence of chymostatin. These findings demonstrate that a dual pathway for the synthesis of angiotensin II is active in diseased and non-diseased coronary arteries. PMID- 15792928 TI - Angiotensin II type 2 receptor-mediated vasodilation. Focus on bradykinin, NO and endothelium-derived hyperpolarizing factor(s). AB - Angiotensin (Ang) II type 1 (AT(1)) receptors account for the majority of the cardiovascular effects Ang II, including vasoconstriction and growth stimulation. Recent evidence, mainly obtained in animals, suggests that Ang II type 2 (AT(2)) receptors counteract some or all of these effects. This review summarizes the current knowledge on the vasodilator effects induced by AT(2) receptors in humans and animals, focussing not only on the mediators of this effect, but also on the modulatory role of age, gender, and endothelial function. It is concluded that AT(2) receptor-mediated vasodilation most likely depends on the bradykinin bradykinin type 2 (B(2)) receptor-NO-cGMP pathway, although evidence for a direct link between AT(2) and B(2) receptors is currently lacking. If indeed B(2) receptors are involved, this would imply that, in addition to NO, also the wide range of non-NO 'endothelium-derived hyperpolarizing factors' (EDHFs) that is released following B(2) receptor activation (e.g., K(+), cytochrome P450 products from arachidonic acid, H(2)O(2) and S-nitrososothiols), could contribute to AT(2) receptor-induced vasodilation. PMID- 15792929 TI - The use of angiotensin-converting enzyme inhibitors in patients undergoing coronary artery bypass graft surgery. AB - Angiotensin-converting enzyme inhibitors have been shown to prolong survival, decrease infarct size, and improve ventricular function in patients who have congestive heart failure, and in those who have sustained a myocardial infarction. This review will summarize the evidence to show that angiotensin converting enzyme inhibitors can also benefit patients undergoing coronary artery bypass graft surgery by minimizing perioperative ischemia, and reducing long-term cardiovascular events. PMID- 15792930 TI - Clinical impact of vasomotor function assessment and the role of ACE-inhibitors and statins. AB - Impaired endothelial function is recognised as one of the earliest events of atherogenesis. Endothelium-dependent vasomotion has been the principal method to assess endothelial function. In this article, we will discuss the clinical value of the different techniques to evaluate endothelium-dependent vasomotion. To date, there seems not to be a simple and reliably endothelial function test to identify asymptomatic subjects at increased risk for cardiovascular disease in clinical practice. Recent studies indicate that pharmacological interventions, in particular with ACE-inhibitors and statins, might improve endothelial function. However, there is no solid evidence that improvement of endothelial function is a necessity for the observed reduction in cardiovascular events by these compounds. Overall, at this moment, there is no place in clinical practice for the use of endothelial function as a method for risk assessment or target of pharmacological interventions. PMID- 15792932 TI - Simulation of minimally invasive vascular interventions for training purposes. AB - OBJECTIVE: To master the skills required to perform minimally invasive vascular interventions, proper training is essential. A computer simulation environment has been developed to provide such training. The simulation is based on an algorithm specifically developed to simulate the motion of a guide wire--the main instrument used during these interventions--in the human vasculature. In this paper, the design and model of the computer simulation environment is described and first results obtained with phantom and patient data are presented. MATERIALS AND METHODS: To simulate minimally invasive vascular interventions, a discrete representation of a guide wire is used which allows modeling of guide wires with different physical properties. An algorithm for simulating the propagation of a guide wire within a vascular system, on the basis of the principle of minimization of energy, has been developed. Both longitudinal translation and rotation are incorporated as possibilities for manipulating the guide wire. The simulation is based on quasi-static mechanics. Two types of energy are introduced: internal energy related to the bending of the guide wire, and external energy resulting from the elastic deformation of the vessel wall. RESULTS: A series of experiments were performed on phantom and patient data. Simulation results are qualitatively compared with 3D rotational angiography data. CONCLUSIONS: The results indicate plausible behavior of the simulation. PMID- 15792933 TI - 3D finite element simulation of Cotrel-Dubousset correction. AB - The Cotrel-Dubousset (CD) scoliosis surgery was simulated for 10 patients with idiopathic scoliosis using a 3D finite element model (FEM) of the patient's entire spine. The geometry of the FEM was extracted from a 3D stereo-radiographic reconstruction, and mechanical properties were personalized using lateral bending films. Finally, each step of the CD correction was simulated and results were compared with the post-operative 3D stereo-radiographic reconstruction. The whole procedure was applied for 10 patients, and quantitative comparison was performed between post-operative spine configuration and predicted configuration. For all patients, mean differences between post-operative measurements and predicted values of vertebral rotation were estimated at 5 degrees (max: 13 degrees) and those for linear position at 6 mm (max: 12 mm). Furthermore, intermediate steps of surgery simulation were consistent with the literature. Then, for one scoliotic patient, the model was used to investigate three alternative surgical strategies. It was found that a one-level change in the instrumentation limit may have a significant effect on spine alignment and correction. PMID- 15792934 TI - C-arm-based mobile computed tomography: a comparison with established imaging on the basis of simulated treatments of talus neck fractures in a cadaveric study. AB - OBJECTIVE: To analyse the image quality and diagnostic effectiveness of a new C arm-based 3D imaging method (C-arm-CT) for intraoperative evaluation of screw osteosyntheses adjacent to a peripheral joint. MATERIALS AND METHODS: Insertion of screws into four cadaveric specimens simulated the surgical treatment of talus neck fractures. Ten orthopedic surgeons and 10 radiologists evaluated X-ray, C arm fluoroscopy, C-arm-CT and CT images. RESULTS: The best image quality was obtained with X-rays (p < 0.001), followed by C-arm fluoroscopy (2D) and CT, with the C-arm-CT (3D) being rated lowest (p < 0.001). The most correct diagnoses were obtained with CT and C-arm-CT (with no statistical difference between them), while C-arm-fluoroscopy was inferior (p < 0.001) and X-rays were the worst (p < 0.05). CONCLUSIONS: Even if the image quality of C-arm-CT is definitely inferior to that of CT, screw misplacements can be reliably detected using C-arm-CT. As compared to the current standard procedures (intraoperative fluoroscopy and postoperative radiography), C-arm-CT performed better. C-arm-CT is ideally suited to the intraoperative diagnosis of high-contrast inquiries like bone fragments and OS material, especially at the extremities. Coupling of the new 3D imaging to existing navigation systems is possible. C-arm-CT will support the further development and implementation of open and minimally invasive surgical procedures. PMID- 15792935 TI - The use of computerized navigation in the treatment of gunshot and shrapnel injury. AB - OBJECTIVE: Trauma surgeons encounter numerous penetrating injuries nowadays. In some cases, missiles causing infection, pain and discomfort, or those retained within joints, bursae and other strategic sites, must be removed. This paper describes an innovative high-tech modality for use in the immediate removal of shrapnel and bullets from strategic anatomical sites. METHODS: Surgical computerized navigation based on real-time acquisition of fluoroscopic data was employed. Several fluoroscopic images of the required anatomical site were obtained. The accurate spatial location of the foreign object could be seen on the images displayed on the computer screen. No further fluoroscopic radiation was necessary. During surgery, the infra-red camera tracked the position of a surgical probe on the patient's anatomy and continuously updated its three dimensional position simultaneously on all displayed images until the missile's location was reached. RESULTS: The use of percutaneous fluoroscopic navigation to remove retained metal objects, including bullets and shrapnel, has proved itself in 12 cases as an accurate measure involving reduced exposure to radiation. In contrast to CT- or MRI-based navigation, computerized fluoroscopic navigation does not require long preliminary preparation. Thus, it is highly efficient in the treatment of acute trauma victims. CONCLUSIONS: The use of this accurate technique in complex and dangerous situations where the foreign body is located in proximity to blood vessels, nerves and narrow 'safe-zones', is promising. This innovative technique reduces surgical time and radiation exposure. In our experience, it has rendered percutaneous missile removal much safer, even in hazardous situations. PMID- 15792936 TI - NEUROGATE: a new MR-compatible device for realizing minimally invasive treatment of intracerebral tumors. AB - The authors report on the handling and the practicability of a newly developed MR compatible device, the NEUROGATE (Daum GmbH, Germany), which allows precise planning, simulation and control of stereotactic biopsy in patients with suspect intracranial lesions, and which allows minimally invasive maneuvers to be performed in a comfortable way. Twenty-eight patients were examined stereotactically in the Signa SP interventional 0.5 Tesla MRI (General Electric Medical Systems, USA), including 15 patients with malignant intracerebral tumors and poor general medical conditions (8 gliomas, 7 metastases) who were treated by laser-induced interstitial thermotherapy (LITT) after definite intraoperative neuropathological diagnosis. As a special stereotactic holding device, the NEUROGATE was favored as a reliable tool for stereotaxy and minimally invasive procedures. PMID- 15792937 TI - 3D reconstruction of the proximal femur with low-dose digital stereoradiography. AB - OBJECTIVE: Accurate three-dimensional (3D) geometry of the proximal femur may be helpful for fracture risk evaluation, as well as for planning and assisting surgical procedures. The purpose of this study was to apply and validate a stereoradiographic 3D reconstruction method on the proximal femur from radiographic contours identified on bi-planar radiographs. MATERIALS AND METHODS: Twenty-five excised non-pathologic proximal femurs were investigated using a low dose digital radiographic device. Three-dimensional personalized models were reconstructed using the Non-Stereo Corresponding Contours (NSCC) algorithm. Three dimensional CT-scan reconstructions were defined as geometric references for the comparison protocol, in order to assess the accuracy and reproducibility of the personalized 3D stereoradiographic reconstructions. In addition, the reliability of a set of 3D parameters obtained from stereoradiographic models was evaluated. RESULTS: This study demonstrated the validity of the NSCC method when applied to the proximal femur, with good results for accuracy (mean error = 0.7 mm) and reproducibility (Wilcoxon test: p > 0.28). Moreover, a precision study for the set of 3D parameters yielded a coefficient of variation lower than 5%. CONCLUSIONS: Once this approach has been validated in vivo, it should find multiple applications in therapeutic fields (e.g., for surgical planning, computer assisted surgery, etc.), as well as in diagnostic contexts (e.g., equilibrium studies or osteoporosis fracture risk assessment). PMID- 15792938 TI - Assessment of video tracking usability for training simulators. AB - OBJECTIVE: A simulator was developed to mimic commercial CAS systems in implementing most tasks required to carry out a surgical operation. As tracking systems are generally expensive components, an alternative solution based on low cost video-based tracking was used. Video tracking accuracy was assessed to determine whether or not this kind of approach was suitable for use in the training domain. Ultimately, video-based tracking should enable sufficiently accurate registration between a bony model and its virtual 3D representation. MATERIALS AND METHODS: Video tracking was assessed using two types of camera. For each one, common accuracy tests were realized as a series of 10 trials at ranges of 0.5-1.0 m from the camera lens. The pointer used as a digitizer was equipped with tracked video markers. Three sizes of marker were evaluated to estimate the impact of marker size on accuracy. RESULTS: For the better of the two cameras tested, results were encouraging. Results are presented as rounded whole-number values in millimeters. The noise test gave accuracies of 2 mm for the 80-mm marker, 3 mm for the 60-mm marker and 5 mm for the 40-mm marker. Relative accuracies, as evaluated on a grid of equally spaced dots, were 4 mm with the 80 mm marker, 7 mm with the 60-mm marker and 12 mm with the 40-mm marker. A pivoting test around the pointer tip gave 3 mm of accuracy for the 80-mm marker, 5 mm for the 60-mm marker and 11 mm for the 40-mm marker. An additional pivoting test was completed on increasing the distance of the marker from the pointer tip, giving accuracies of 5 mm for the 80-mm marker, 6 mm for the 60-mm marker and 13 mm for the 40-mm marker. The registration test gave accuracies of 8 mm for the 80-mm marker, 9 mm for the 60-mm marker and 11 mm for the 40-mm marker. CONCLUSIONS: The video-based approach offers sufficient accuracy to achieve registration in the domain of CAS training. PMID- 15792939 TI - Evaluation of deformable models for femoral neck surgery. AB - OBJECTIVE: Using fluoroscopic images alone, it is difficult to guarantee that screws are positioned within the femoral head and neck. This study evaluates whether the introduction of deformable 3D models limiting the planning and navigation space is a helpful approach to minimizing the incidence of misplaced screws, thereby enhancing patient safety. BACKGROUND: Even though a screw may appear to lie within the femoral head and neck on fluoroscopic images, this may not, in fact, be the case. This is a particular problem for interventions such as fixation of a slipped femoral head or osteosynthesis of the femoral neck, where screws must be set close to the cortical bone without penetrating the joint or injuring the cortex of the femoral neck. METHODS: A system was developed which permits computer-based planning and navigation of screws for femoral neck fracture fixation based on fluoroscopic images. Different approaches were employed which either a) make use of a deformable model adapted to the femoral head/neck, constraining the screw positions within this model; or b) allow the user to position the screws with or without geometrical constraints on the X-rays while maintaining parallelism of the screws. All designs were evaluated and compared by 7 test users using integral projection X-rays calculated from the CT dataset. Results were checked using a 3D model of the bone, also calculated from the CT dataset. RESULTS: Positioning screws using the deformable model resulted in a significantly smaller distribution of screw tip locations and no penetrations into the hip joint, in contrast to the other approaches where up to 11% of screws were misplaced. CONCLUSIONS: Constraining the planning and navigation space by means of a deformable model allows better control of screw positioning and thus increases the chances of a successful intervention. In particular, CAS systems allowing for virtual fluoroscopy should consider supporting this planning approach. PMID- 15792940 TI - Computer-enhanced insertion of the Oxford unicompartmental arthroplasty: a fluoroguide technique. AB - OBJECTIVE: To develop an accurate and reproducible technique for inserting Oxford Unicompartmental arthroplasties. MATERIALS AND METHODS: A protocol was developed to accurately position the femoral component of the Oxford Unicompartmental arthroplasty using computer-enhanced techniques. A dynamic reference body (DRB) is inserted into the femur, then antero-posterior (AP) and lateral fluoroscopic images are taken of the proximal femur. These images are digitized and entered into 3D space using a registered fluoroscopy machine. The femoral guide from the Oxford system is then placed on the medial femoral condyle in the standard manner. Holes are drilled directly towards the center of the femoral head using a registered drill. The femoral cuts are then made using the standard instruments from the Oxford set. RESULTS: This procedure was carried out on 15 sawbone femurs. Component position was determined by measuring radiographs after a trial component had been inserted. The average lateral error was 1.2 degrees (standard deviation [SD] = 1.207), compared to 4.1 degrees (SD = 2.875) with the standard technique. The average AP error was 2.5 degrees (SD = 1.767), compared to 5.1 degrees (SD = 2.219) with the standard technique. The differences in accuracy in both planes were statistically significant (p = 0.002 for AP, 0.001 for lateral). CONCLUSION: Using a Fluoroguide-assisted technique, we were able to insert the femoral component more precisely than when using the standard technique as described by the manufacturer. PMID- 15792943 TI - Endocannabinoids and food intake: newborn suckling and appetite regulation in adulthood. AB - The appetite-stimulating effects of the cannabis plant (Cannabis sativa) have been known since ancient times, and appear to be effected through the incentive and rewarding properties of foods. Investigations into the biological basis of the multiple effects of cannabis have yielded important breakthroughs in recent years: the discovery of two cannabinoid receptors in brain and peripheral organ systems, and endogenous ligands (endocannabinoids) for these receptors. These advances have greatly increased our understanding of how appetite is regulated through these endocannabinoid receptor systems. The presence of endocannabinoids in the developing brain and in maternal milk have led to evidence for a critical role for CB1 receptors in oral motor control of suckling during neonatal development. The endocannabinoids appear to regulate energy balance and food intake at four functional levels within the brain and periphery: (i) limbic system (for hedonic evaluation of foods), (ii) hypothalamus and hindbrain (integrative functions), (iii) intestinal system, and (iv) adipose tissue. At each of these levels, the endocannabinoid system interacts with a number of better known molecules involved in appetite and weight regulation, including leptin, ghrelin, and the melanocortins. Therapeutically, appetite stimulation by cannabinoids has been studied for several decades, particularly in relation to cachexia and malnutrition associated with cancer, acquired immunodeficiency syndrome, or anorexia nervosa. The recent advances in cannabinoid pharmacology may lead to improved treatments for these conditions or, conversely, for combating excessive appetite and body weight, such as CB1 receptor antagonists as antiobesity medications. In conclusion, the exciting progress in the understanding of how the endocannabinoid CB receptor systems influence appetite and body weight is stimulating the development of therapeutic orexigenic and anorectic agents. Furthermore, the role of cannabinoid CB1 receptor activation for milk suckling in newborns may open new doors toward understanding nonorganic failure-to-thrive in infants, who display growth failure without known organic cause. PMID- 15792944 TI - Gamma-tocotrienol inhibits neoplastic mammary epithelial cell proliferation by decreasing Akt and nuclear factor kappaB activity. AB - Tocotrienols, a subgroup within the vitamin E family of compounds, have been shown to display potent anticancer activity and inhibit preneoplastic and neoplastic mammary epithelial cell proliferation at treatment doses that have little or no effect on normal cell growth and function. However, the specific intracellular mechanisms mediating the antiproliferative effects of tocotrienols are presently unknown. Because Akt and nuclear factor kappaB (NFkappaB) are intimately involved in mammary tumor cell proliferation and survival, studies were conducted to determine the effects of gamma-tocotrienol on Akt and NFkappaB activity in neoplastic +SA mammary epithelial cells in vitro. Treatment with 0-8 microM gamma-tocotrienol for 0-3 days caused a dose-responsive inhibition in +SA cell growth and mitotic activity, as determined by MTT colorimetric assay and proliferating cell nuclear antigen immunocytochemical staining, respectively. Studies also showed that treatment with 4 microM gamma-tocotrienol, a dose that inhibited +SA cell growth by more than 50% compared with that of untreated control cells, decreased intracellular levels of activated phosphotidylinositol 3 kinase-dependent kinase (PI3K)-dependent kinase 1 (phospho-PDK-1) and Akt, and reduced phospho-Akt kinase activity. Furthermore, these effects were not found to be associated with an increase in either phosphatase and tensin homologue deleted from chromosome 10 (PTEN) or protein phosphatase type 2A phosphatase activity. In addition, gamma-tocotrienol treatment was shown to decrease NFkappaB transcriptional activity, apparently by suppressing the activation of IkappaB kinase-alpha/beta, an enzyme associated with inducing NFkappaB activation. In summary, these findings demonstrate that the antiproliferative effects of gamma tocotrienol result, at least in part, from a reduction in Akt and NFkappaB activity in neoplastic +SA mammary epithelial cells. PMID- 15792946 TI - Serum leptin in nonpregnant and pregnant women and in old and new world nonhuman primates. AB - Leptin is a hormone that is produced during mammalian pregnancy in the placental trophoblast and other tissues, including! fetal and maternal adipocytes. Synthesis of the polypeptide and the presence of its specific receptors throughout the human maternal fetoplacental unit suggest direct effects on conceptus growth and development. However, both the physiologic roles of leptin and the mechanisms regulating leptin synthesis in human pregnancy differ from those in laboratory and domestic species, necessitating the development of non human primate research models. Therefore, we compared serum leptin concentrations in nonpregnant and pregnant women with those in both old world nonhuman primates (i.e., baboon, rhesus monkey, cynomolgus monkey) and new world nonhuman primates (i.e., squirrel monkey, titi monkey). As expected, maternal leptin levels were elevated in human and baboon pregnancies (P < 0.05 and P < 0.001, respectively). Levels in both species of old world monkeys were also greatly enhanced (P < 0.001). Although maternal serum concentrations were slightly elevated compared to nonpregnant levels in both species of new world monkeys, overall concentrations were dramatically lower than for either old world primates or humans. Results provide comparisons of serum leptin concentrations in pregnant and nonpregnant humans and baboons with those in both old and new world monkeys and further characterize these nonhuman primates as models for the investigation of leptin dynamics in pregnancy. PMID- 15792947 TI - Effects of chondroitin and glucosamine sulfate in a dietary bar formulation on inflammation, interleukin-1beta, matrix metalloprotease-9, and cartilage damage in arthritis. AB - This study examined the effects of chondroitin sulfate (CS) alone and CS plus glucosamine sulfate (GS) in a dietary bar formulation on inflammatory parameters of adjuvant-induced arthritis and on the synthesis of interleukin-1beta (IL 1beta) and matrix metalloprotease-9 (MMP-9). Following 25 days pretreatment with dietary bars containing either CS alone, CS plus GS, or neither CS nor GS, rats were either sham injected or injected with Freund's complete adjuvant into the tail vein. Rats were fed their respective bars for another 17 days after inoculation. Parameters of disease examined included clinical score (combination of joint temperature, edema, hyperalgesia, and standing and walking limb function), incidence of disease, levels of IL-1beta in the serum and paw joints, levels of MMP-9 in the paw joints, paw joint histology, and joint cartilage thickness. Treatment with CS plus GS, but not CS alone, significantly reduced clinical scores, incidences of disease, joint temperatures, and joint and serum IL-1beta levels. Treatment with CS alone and CS plus GS inhibited the production of edema and prevented raised levels of joint MMP-9 associated with arthritis. Similarly, CS alone and CS plus GS treatment also prevented the development of cartilage damage associated with arthritis. Combination CS plus GS treatment in a dietary bar formulation ameliorates clinical, inflammatory, and histologic parameters of adjuvant-induced arthritis. The benefits of CS and GS in combination are more pronounced than those of CS alone. The reduction of arthritic disease by CS plus GS is associated with a reduction of IL-1beta and MMP-9 synthesis. PMID- 15792945 TI - Differential effects of cGMP produced by soluble and particulate guanylyl cyclase on mouse ventricular myocytes. AB - Particulate guanylyl cyclase (pGC) and soluble guanylyl cyclase (sGC) are cGMP generation systems distributed in different intracellular locations. Our aim was to test the hypothesis that the functional effects of cGMP produced by pGC and sGC on contraction and Ca2+ transients would differ in ventricular myocytes. We measured myocyte shortening from adult mice using a video edge-detector and investigated the functional changes after stimulating pGC with C-type natriuretic peptide (CNP; 10(-8) M and 10(-7) M) or sGC with S-nitroso-N-acetyl-penicillamine (SNAP; nitric oxide donor; 10(-6) M and 10(-5) M). Significant concentration dependent decreases in percentage shortening (PCS), maximal rate of shortening (RSmax), and relaxation (RRmax) were produced by CNP. To a similar degree, SNAP concentration-dependently reduced PCS, RSmax, and RRmax. The addition of Rp-8-[(4 chlorophenyl)thio]-cGMPS triethylamine (cGMP-dependent protein kinase inhibitor; 5 x 10(-6) M) or erythro-9-(2-hydroxy-3-nonyl) adenine (cGMP-stimulated cAMP phosphodiesterase inhibitor; 10(-5) M) reduced the responses induced by CNP or SNAP, suggesting that their actions were through cGMP-mediated pathways. While SNAP significantly increased intracellular cGMP concentration by 57%, CNP had little effect on cGMP production. We also found that CNP markedly decreased the amplitude of Ca2+ transients while SNAP had little effect, suggesting the cGMP generated by sGC may decrease myofilament Ca2+ sensitivity. The small amount of cGMP generated by pGC had a major effect in reducing Ca2+ level. This study suggested the existence of compartmentalization for cGMP in ventricular myocytes. PMID- 15792948 TI - The role of human telomerase catalytic subunit mRNA expression in cervical dysplasias. AB - Telomerase activity and human telomerase reverse transcriptase (hTERT) mRNA expression were investigated in cervical specimens and were correlated with cytologic findings and the presence of human papilloma virus (HPV) infection. Telomerase activity was evaluated by the telomeric repeat protocol assay and hTERT mRNA expression was evaluated by reverse transcriptase polymerase chain reaction (PCR). HPV DNA was detected by PCR, as well as restriction endonuclease digestion. HPV DNA was detected in all 82 specimens with abnormal cytologic findings and in 4 of 34 normal samples. Low-grade squamous intraepithelial lesions (LGSILs) were present in 74 of 82 specimens (90.2%) and high-grade squamous intraepithelial lesions (HGSILs) were present in 8 of 82 (9.75%) specimens. Seven of the eight HGSIL (87.5%) and 26 of 74 LGSIL (35.1%) specimens were hTERT positive, whereas all normal specimens were hTERT mRNA negative. Telomerase activity was detected in 21 of 74 (28.4%) LGSIL/atypical squamous epithelial cells of undetermined significance (ASCUS) and in five of eight (62.5%) HGSIL samples. A correlation was observed among telomerase activity, hTERT mRNA expression, and high-risk HPV infection in HGSIL samples (P < 0.001). High-risk HPV infection assessment showed 75% sensitivity and 72.2% specificity for HGSILs. Telomerase activity assessment in cervical smears showed sensitivity and negative predictive value (NPV) for HGSILs 62.5% and 96.7%, whereas specificity and positive predictive value (PPV) were 80.5% and 19.2%, respectively. hTERT mRNA expression assessment showed 87.5% sensitivity and 98.7% NPV for HGSILs, whereas specificity and PPV were 76% and 21.2%, respectively. Based on the above-described telomerase assessment values, it is suggested that the telomerase system might not be an appropriate diagnostic marker for cytology, given that the final evaluation must rely on a combination of all available test assessment data, clinical diagnosis, as well as the follow-up of all LGSIL samples that were positive for telomerase activation. PMID- 15792949 TI - Exogenous administration of Substance P enhances wound healing in a novel skin injury model. AB - Soft tissue injury accounts for approximately 44% of all wounds in both the military and civilian populations. Following injury to soft tissue, Substance P (SP) and other neuropeptides are released by cutaneous neurons and modulate the function of immunocompetent and inflammatory cells, as well as epithelial and endothelial cells. The interaction between these components of the nervous system and multiple target cells affecting cutaneous repair is of increasing interest. In this report, we describe the effects of SP on wound repair in a novel, laser induced, skin-wound model. Gross and histologic examination of laser-induced injury revealed that exogenously administered SP affects wound healing via neurite outgrowth, in addition to adhesion molecule and neurokinin-1 receptor involvement in vivo. All SP effects were decreased by pretreatment with Spantide II, an SP antagonist. The elucidation of SP-mediating mechanisms is crucial to firmly establishing the involvement and interaction of the peripheral nervous system and the immune system in cutaneous repair. Findings presented here suggest that SP participates in the complex network of mediators involved in cutaneous inflammation and wound healing. PMID- 15792950 TI - Protective role of urinary trypsin inhibitor in acute lung injury induced by lipopolysaccharide. AB - Urinary trypsin inhibitor (UTI), a serine protease inhibitor, has been widely used as a drug for patients with acute inflammatory disorders such as disseminated intravascular coagulation, shock, and pancreatitis. However, direct contribution of UTI to inflammatory diseases has not been established. The present study analyzed acute inflammatory lung injury induced by lipopolysaccharide (LPS) in UTI-deficient (-/-) mice and corresponding wild-type (WT) mice. UTI (-/-) and WT mice were treated intratracheally with vehicle or LPS (125 mug/kg). The cellular profile of bronchoalveolar lavage fluid, lung water content, histology, and expression of proinflammatory molecules in the lung were evaluated. After LPS challenge, both genotypes of mice revealed neutrophilic lung inflammation and pulmonary edema. UTI (-/-) mice, however, showed more prominent infiltration of inflammatory cells and edema than WT mice. After LPS challenge in both genotypes of mice, the lung levels of mRNA and/or protein expression of interleukin-1beta, macrophage inflammatory protein-1alpha, macrophage chemoattractant protein-1, keratinocyte chemoattractant, and intercellular adhesion molecule-1 (ICAM-1) were elevated in both groups, but to a greater extent in UTI (-/-) mice than in WT mice. These results suggest that UTI protects against acute lung injury induced by bacterial endotoxin, at least partly, through the inhibition of the enhanced local expression of proinflammatory cytokines, chemokines, and ICAM-1. PMID- 15792951 TI - Switching human telomerase on and off with hPOT1 protein in vitro. AB - POT1 (protection of telomeres 1) protein binds the G-rich single-stranded telomeric DNA at the ends of chromosomes. In human cells hPOT1 is involved in telomere length regulation, but the mechanism of this regulation remains unknown. Examination of the high-resolution crystal structure of the hPOT1-TTAGGGTTAG complex suggested that it would not be extended by telomerase, a hypothesis that we confirm by in vitro assays with recombinant telomerase. On the other hand, when hPOT1 is bound at a position one telomeric repeat before the 3'-end, leaving an 8-nucleotide 3'-tail, the complex is extended with improved activity and processivity. Thus, depending on its location relative to the DNA 3'-end, hPOT1 can either inhibit telomerase action or form a preferred substrate for telomerase. We propose that another factor catalyzes the interconversion of these states in vivo. PMID- 15792952 TI - Mechanistic studies on a novel, highly potent gold-phosphole inhibitor of human glutathione reductase. AB - The homodimeric flavoprotein glutathione reductase (GR) is a central player of cellular redox metabolism, connecting NADPH to the large pool of redox-active thiols. In this work, the inhibition of human GR by a novel gold-phosphole inhibitor (GoPI) has been studied in vitro. Two modes of inhibition are observed, reversible inhibition that is competitive with GSSG followed by irreversible inhibition. When approximately 1 nm GoPI is incubated with NADPH-reduced GR (1.4 nm) the enzyme becomes 50% inhibited. This appears to be the most potent stable inhibitor of human GR to date. Analyzing the monophasic oxidative half-reaction of reduced GR with GSSG at pH 6.9 revealed a K(d)((app)) for GSSG of 63 microm, and a k((obs)max) of 106 s(-1) at 4 degrees C. The reversible inhibition by the gold-phosphole complex [{1-phenyl-2,5-di(2-pyridyl)phosphole}AuCl] involves formation of a complex at the GSSG-binding site of GR (K(d) = 0.46 microm) followed by nucleophilic attack of an active site cysteine residue that leads to covalent modification and complete inactivation of the enzyme. Data from titration spectra, molecular modeling, stopped-flow, and steady-state kinetics support this theory. In addition, covalent binding of the inhibitor to human GR was demonstrated by mass spectrometry. The extraordinary properties of the compound and its derivatives might be exploited for cell biological studies or medical applications, e.g. as an anti-tumor or antiparasitic drug. Preliminary experiments with glioblastoma cells cultured in vitro indicate an anti proliferative effect of the inhibitor in the lower micromolar range. PMID- 15792953 TI - Cyanobacterial non-mevalonate pathway: (E)-4-hydroxy-3-methylbut-2-enyl diphosphate synthase interacts with ferredoxin in Thermosynechococcus elongatus BP-1. AB - (E)-4-Hydroxy-3-methylbut-2-enyl diphosphate synthase (GcpE), which catalyzes the conversion of 2-C-methyl-D-erythritol cyclodiphosphate (MEcPP) into (E)-4-hydroxy 3-methylbut-2-enyl diphosphate (HMBPP), is an essential enzyme of the non mevalonate (2-C-methyl-D-erythritol-4-phosphate (MEP)) pathway for isoprenoid biosynthesis. The terminal steps of the MEP pathway are still not fully understood, although this pathway is necessary for survival in various organisms such as cyanobacteria, plastids of algae and higher plants, and the apicoplast of human malaria parasites. To determine the efficient redox partner for thermophilic cyanobacterial GcpE, We have expressed the gcpE and petF genes in Escherichia coli and studied the protein-protein interaction of GcpE protein with ferredoxin I (PetF) from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1. Recombinant GcpE protein was purified by an N-terminal His(6) tag and reconstituted as a [4Fe-4S](2+) metalloprotein. GcpE was shown to interact strongly with PetF via the bacterial two-hybrid system designed to detect protein protein interactions. Moreover, a direct protein-protein interaction between PetF and GcpE was confirmed in an in vitro glutathione S-transferase (GST) pull-down assay. To investigate electron transfer activity from PetF to GcpE, we also constructed a NADPH-dependent reducing shuttle system with purified recombinant ferredoxin-NADP(+) oxidoreductase (PetH) and PetF. The result demonstrated that PetF has the ability to transfer electrons to GcpE. Thus, the combined data provide the first evidence that GcpE is a ferredoxin-dependent enzyme in T. elongatus BP-1. PMID- 15792954 TI - Properties of the permeability transition pore in mitochondria devoid of Cyclophilin D. AB - We have studied the properties of the permeability transition pore (PTP) in mitochondria from the liver of mice where the Ppif gene encoding for mitochondrial Cyclophilin D (CyP-D) had been inactivated. Mitochondria from Ppif /- mice had no CyP-D and displayed a striking desensitization of the PTP to Ca2+, in that pore opening required about twice the Ca2+ load necessary to open the pore in strain-matched, wild-type mitochondria. Mitochondria lacking CyP-D were insensitive to Cyclosporin A (CsA), which increased the Ca2+ retention capacity only in mitochondria from wild-type mice. The PTP response to ubiquinone 0, depolarization, pH, adenine nucleotides, and thiol oxidants was similar in mitochondria from wild-type and Ppif-/- mice. These experiments demonstrate that (i) the PTP can form and open in the absence of CyP-D, (ii) that CyP-D represents the target for PTP inhibition by CsA, and (iii) that CyP-D modulates the sensitivity of the PTP to Ca2+ but not its regulation by the proton electrochemical gradient, adenine nucleotides, and oxidative stress. These results have major implications for our current understanding of the PTP and its modulation in vitro and in vivo. PMID- 15792955 TI - Coq3 and Coq4 define a polypeptide complex in yeast mitochondria for the biosynthesis of coenzyme Q. AB - Coenzyme Q (Q) is a redox active lipid essential for aerobic respiration in eukaryotes. In Saccharomyces cerevisiae at least eight mitochondrial polypeptides, designated Coq1-Coq8, are required for Q biosynthesis. Here we present physical evidence for a coenzyme Q-biosynthetic polypeptide complex in isolated mitochondria. Separation of digitonin-solubilized mitochondrial extracts in one- and two-dimensional Blue Native PAGE analyses shows that Coq3 and Coq4 polypeptides co-migrate as high molecular mass complexes. Similarly, gel filtration chromatography shows that Coq1p, Coq3p, Coq4p, Coq5p, and Coq6p elute in fractions higher than expected for their respective subunit molecular masses. Coq3p, Coq4p, and Coq6p coelute with an apparent molecular mass exceeding 700 kDa. Coq3 O-methyltransferase activity, a surrogate for Q biosynthesis and complex activity, also elutes at this high molecular mass. We have determined the quinone content in lipid extracts of gel filtration fractions by liquid chromatography-tandem mass spectrometry and find that demethoxy-Q(6) is enriched in fractions with Coq3p. Co-precipitation of biotinylated-Coq3 and Coq4 polypeptide from digitonin-solubilized mitochondrial extracts shows their physical association. This study identifies Coq3p and Coq4p as defining members of a Q-biosynthetic Coq polypeptide complex. PMID- 15792956 TI - Silibinin up-regulates DNA-protein kinase-dependent p53 activation to enhance UVB induced apoptosis in mouse epithelial JB6 cells. AB - In the present study, we employed a well established JB6 mouse epithelial cell model to define the molecular mechanism of efficacy of a naturally occurring flavonoid silibinin against ultraviolet B (UVB)-induced skin tumorigenesis. UVB exposure of cells caused a moderate phosphorylation of ERK1/2 and Akt and a stronger phosphorylation of p53 at Ser(15), which was enhanced markedly by silibinin pretreatment. Kinase activity of ERK1/2 for Elk-1 and Akt for glycogen synthase kinase-3beta was also potently enhanced by silibinin pretreatment. Furthermore, silibinin increased the UVB-induced level of cleaved caspase 3 as well as apoptotic cells. Based on these observations, next we investigated the role of upstream kinases, ATM/ATR and DNA-PK, which act as sensors for UVB induced DNA damage and transduce signals leading to DNA repair or apoptosis. Whereas UVB strongly activated ATM as observed by Ser(1981) phosphorylation, it was not affected by silibinin pretreatment. However, pretreatment of cells with the DNA-protein kinase (PK) inhibitor LY294002 strongly reversed silibinin enhanced Akt-Ser(473) and p53-Ser(15) as well as ERK1/2 phosphorylation together with a dose-dependent decrease in cleaved caspase 3 and apoptosis (p < 0.05). In addition, silibinin pretreatment strongly enhanced H2A.X-Ser(139) phosphorylation and DNA-PK-associated kinase activity as well as the physical interaction of p53 with DNA-PK; pretreatment of cells with LY294002 but not caffeine abolished the silibinin-caused increase in both DNA-PK activation and p53-Ser(15) phosphorylations. Together, these findings suggest that silibinin preferentially activates the DNA-PK-p53 pathway for apoptosis in response to UVB-induced DNA damage, and that this could be a predominant mechanism of silibinin efficacy against UVB-induced skin cancer. PMID- 15792957 TI - Activation of the rat renin promoter by HOXD10.PBX1b.PREP1, Ets-1, and the intracellular domain of notch. AB - Renin gene expression is subject to complex developmental and tissue-specific regulation. A comparison of the promoter sequences of the human, rat, and mouse renin genes has revealed a highly conserved sequence homologous to the DNA recognition sequence for CBF1 (CSL/RBP-Jkappa/Su(H)/LAG1/RBPSUH). Electrophoretic mobility shift assays document that As4.1 cell nuclear protein complex binding to the putative rat renin CBF1-binding site (-175 to -168 bp) contains CBF1. Transient transfection analyses in COS-7 cells further document that a CBF1-VP16 fusion protein and the intracellular domain of Notch1 robustly activate a promoter containing multiple copies of the rat renin CBF1-binding site. An Ets binding site (-143 to -138 bp) has also been identified in the rat renin promoter by sequence comparisons and electrophoretic mobility shift assays. Transcription factor Ets-1 is capable of activating the rat renin promoter through the Ets binding site. Mutation of the CBF-binding site significantly increases transcriptional activity of the rat renin promoter in Calu-6 and COS-7 cells but not in As4.1 cells, whereas mutation of the Ets-binding site reduces promoter activity of the rat renin gene in all three cell lines. Finally, we show that the intracellular domain of Notch1, Ets-1, and HOXD10.PBX1b.PREP1 activate the rat renin promoter cooperatively in COS-7 cells. These results strongly suggest that the renin gene is a downstream target of the Notch signaling pathway. PMID- 15792959 TI - Molecular characterization and origin of novel bipartite cold-regulated ice recrystallization inhibition proteins from cereals. AB - To understand the molecular basis of freezing tolerance in plants, several low temperature-responsive genes have been identified from wheat. Among these are two genes named TaIRI-1 and TaIRI-2 (Triticum aestivum ice recrystallization inhibition) that are up-regulated during cold acclimation in freezing-tolerant species. Phytohormones involved in pathogen defense pathways (jasmonic acid and ethylene) induce the expression of one of the two genes. The encoded proteins are novel in that they have a bipartite structure that has never been reported for antifreeze proteins. Their N-terminal part shows similarity with the leucine-rich repeat-containing regions present in the receptor domain of receptor-like protein kinases, and their C-terminus is homologous to the ice-binding domain of some antifreeze proteins. The recombinant TaIRI-1 protein inhibits the growth of ice crystals, confirming its function as an ice recrystallization inhibition protein. The TaIRI genes were found only in the species belonging to the Pooideae subfamily of cereals. Comparative genomic analysis suggested that molecular evolutionary events took place in the genome of freezing-tolerant cereals to give rise to these genes with putative novel functions. These apparent adaptive DNA rearrangement events could be part of the molecular mechanisms that ensure the survival of hardy cereals in the harsh freezing environments. PMID- 15792960 TI - RNAi-mediated silencing of OsGEN-L (OsGEN-like), a new member of the RAD2/XPG nuclease family, causes male sterility by defect of microspore development in rice. AB - We have cloned a new member of the RAD2/XPG nuclease family, OsGEN-L (OsGEN like), from rice (Oryza sativa L.). OsGEN-L possesses two domains, the N- and I regions, that are conserved in the RAD2/XPG nuclease family. Database searches and phylogenetic analyses revealed that OsGEN-L belongs to class 4 of the RAD2/XPG nuclease family, and OsGEN-L homologs were found in animals and higher plants. To elucidate the function of OsGEN-L, we generated rice OsGEN-L-RNAi transgenic plants in which OsGEN-L expression was silenced. Most of the OsGEN-L RNAi plants displayed low fertility, and some of them were male-sterile. OsGEN-L RNAi plants lacked mature pollen, resulting from a defect in early microspore development. A OsGEN-L-green fluorescent protein (GFP) fusion protein was localized in the nucleus, and the OsGEN-L promoter was specifically active in the anthers. Furthermore, a recombinant OsGEN-L protein possessed flap endonuclease activity and both single-stranded and double-stranded DNA-binding activities. Our results suggest that OsGEN-L plays an essential role in DNA metabolism required for early microspore development in rice. PMID- 15792961 TI - Peroxisomal localization of a myosin XI isoform in Arabidopsis thaliana. AB - The genome of Arabidopsis thaliana contains 13 myosin XI isoforms. Here we prepared a specific antibody against a peptide that mimics a unique C-terminal region from the myosin XI isoform, MYA2. The resulting antibody was used to demonstrate that MYA2 in Arabidopsis protein extracts co-sedimented with actin filaments and dissociated from the filaments with ATP treatment. Immunolocalization studies showed that MYA2 co-localized predominantly with actin filaments in clustered punctuate dots in leaf epidermal cells, root hair cells and suspension-cultured cells. In a transgenic plant in which peroxisomes are labeled with green fluorescent protein, some MYA2 signals were localized on peroxisomes in an actin-dependent manner. We propose that the peroxisome is one of the cargos translocated by MYA2 on actin filaments. PMID- 15792962 TI - Linkage disequilibrium fine mapping and haplotype association analysis of the tau gene in progressive supranuclear palsy and corticobasal degeneration. AB - BACKGROUND: The haplotype H1 of the tau gene, MAPT, is highly associated with progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). OBJECTIVE: To investigate the pathogenic basis of this association. METHODS: Detailed linkage disequilibrium and common haplotype structure of MAPT were examined in 27 CEPH trios using validated HapMap genotype data for 24 single nucleotide polymorphisms (SNPs) spanning MAPT. RESULTS: Multiple variants of the H1 haplotype were resolved, reflecting a far greater diversity of MAPT than can be explained by the H1 and H2 clades alone. Based on this, six haplotype tagging SNPs (htSNPs) that capture 95% of the common haplotype diversity were used to genotype well characterised PSP and CBD case-control cohorts. In addition to strong association with PSP and CBD of individual SNPs, two common haplotypes derived from these htSNPs were identified that are highly associated with PSP: the sole H2 derived haplotype was underrepresented and one of the common H1 derived haplotypes was highly associated, with a similar trend observed in CBD. There were powerful and highly significant associations with PSP and CBD of haplotypes formed by three H1 specific SNPs. This made it possible to define a candidate region of at least approximately 56 kb, spanning sequences from upstream of MAPT exon 1 to intron 9. On the H1 haplotype background, these could harbour the pathogenic variants. CONCLUSIONS: The findings support the pathological evidence that underlying variations in MAPT could contribute to disease pathogenesis by subtle effects on gene expression and/or splicing. They also form the basis for the investigation of the possible genetic role of MAPT in Parkinson's disease and other tauopathies, including Alzheimer's disease. PMID- 15792963 TI - Cyclic GMP-specific phosphodiesterase 5 regulates growth and apoptosis in pulmonary endothelial cells. AB - Sustained increases in intracellular cGMP concentrations ([cGMP]i) inhibit cell growth and induce apoptosis. We now report that a cGMP-specific phosphodiesterase, PDE5, plays a dominant role in regulating [cGMP]i transitions that inhibit cell growth and control susceptibility to apoptosis in pulmonary endothelium. Atrial natriuretic peptide (ANP) activates guanylyl cyclase A/B and induces a rapid [cGMP]i rise 2-5 min after its application, in both pulmonary arterial endothelial cells (PAECs) and pulmonary microvascular endothelial cells (PMVECs). However, increased [cGMP]i in PAECs is transient and decays within 10 min due to cytosolic PDE5 hydrolytic activity. Increased [cGMP]i in PMVECs is sustained for >3 h due to the absence of PDE5. Indeed, at any ANP concentration, the sustained (30 min) [cGMP]i rise is greater in PMVECs than in PAECs, unless PAECs are also treated with the PDE5 inhibitor zaprinast. Using RT-PCR, Western blot analysis, immunoprecipitation, and DEAE chromatography, we resolved the expression and activity of PDE 5A1/A2 only in PAECs. Similarly, PDE5 expression was restricted to extra-alveolar endothelium in vivo. ANP induced growth inhibition and apoptosis in PMVECs, but similar effects were not seen in PAECs unless ANP treatment was combined with zaprinast. ANP blocked the VEGF-induced proliferation and migration in PMVECs. Collectively, these data suggest that PDE5 regulated [cGMP]i controls endothelial cell growth and apoptosis, representing a mechanism of heterogeneity between two endothelial phenotypes. PMID- 15792964 TI - Protection against bleomycin-induced lung injury by IL-18 in mice. AB - The role of interleukin (IL)-18 in the protection from interstitial pneumonia and pulmonary fibrosis induced by bleomycin (BLM) was investigated by comparing the severity of BLM-induced lung injuries between wild-type and C57BL/6 mice with a targeted knockout mutation of the IL-18 gene (IL-18-/- mice). IL-18-/- mice showed much worse lung injuries than wild-type mice, as assessed by the survival rate, histological images, and leukocyte infiltration in the bronchoalveolar lavage fluid and myeloperoxidase activity. In wild-type mice, administration of IL-18 before BLM instillation resulted in suppression of lung injuries, increases in the hydroxyproline content, and decreases in the granulocyte-macrophage colony stimulating factor content in the lung. Preadministration of IL-18 also resulted in prevention of the reduction of the lung IL-10 content caused by BLM-induced damage of alveolar epithelial. BLM instillation suppressed superoxide dismutase (SOD) activity in IL-18-/- mice to a greater extent than in wild-type mice. Pretreatment of IL-18 augmented Mn-containing superoxide dismutase (Mn-SOD) messenger RNA expression and SOD activity in the lung and prevented the reduction of SOD activity caused by BLM in both wild-type and IL-18-/- mice. These results suggest that IL-18 plays a protective role against BLM-induced lung injuries by upregulating a defensive molecule, Mn-SOD. PMID- 15792965 TI - Activation of the aryl hydrocarbon receptor increases pulmonary neutrophilia and diminishes host resistance to influenza A virus. AB - Unlike their role in bacterial infection, less is known about the role of neutrophils during pulmonary viral infection. Exposure to pollutant 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD, dioxin) results in excess neutrophils in the lungs of mice infected with influenza A virus. TCDD is the most potent agonist for the aryl hydrocarbon receptor (AhR), and exposure to AhR ligands has been correlated with exacerbated inflammatory lung diseases. However, knowledge of the effects of AhR agonists on neutrophils is limited. Likewise, the factors regulating neutrophil responses during respiratory viral infections are not well characterized. To address these knowledge gaps, we determined the in vivo levels of KC, MIP-1alpha, MIP-2, LIX, IL-6, and C5a in infected mouse lungs. Our data show that these neutrophil chemoattractants are generally produced transiently in the lung within 12-24 h of infection. We also report that expression of CD11a, CD11b, CD49d, CD31, and CD38 is increased on pulmonary neutrophils in response to influenza virus. Using AhR-deficient mice, we demonstrate that excess neutrophilia in the lung is mediated by activation of the AhR and that this enhanced neutrophilia correlates directly with decreased survival in TCDD-exposed mice. Although AhR activation results in more neutrophils in the lungs, we show that this is not mediated by deregulation in levels of common neutrophil chemoattractants, expression of adhesion molecules on pulmonary neutrophils, or delayed death of neutrophils. Likewise, exposure to TCDD did not enhance pulmonary neutrophil function. This study provides an important first step in elucidating the mechanisms by which AhR agonists exacerbate pulmonary inflammatory responses. PMID- 15792966 TI - Dopamine type 2 receptor expression and function in rodent sensory neurons projecting to the airways. AB - Agonists of the dopamine receptors have been demonstrated to have bronchodilatory properties in pathologically constricted airways. The mechanism by which these agonists induce bronchodilatation is thought to involve airway sensory nerves. In this study, the expression and function of dopamine D(2) receptor were examined in sensory ganglia supplying the airways. Neuronal dopamine D(2) receptor mRNA expression was demonstrated by single-cell RT-PCR following laser-assisted microdissection. The projection of the neurons to the airways was confirmed by retrograde neuronal labeling. In functional studies, dopamine D(2) receptor agonists (AR-C65116AB and ropinirole) inhibited intraneuronal calcium mobilization in rat capsaicin-sensitive primary sensory neurons and capsaicin induced plasma extravasation in the rat trachea. Our results provide support to the hypothesis that dopamine D(2) receptor activation inhibits neurogenic inflammation and proinflammatory reflex responses. PMID- 15792967 TI - Complex stability of single proteins explored by forced unfolding experiments. AB - In the last decade atomic force microscopy has been used to measure the mechanical stability of single proteins. These force spectroscopy experiments have shown that many water-soluble and membrane proteins unfold via one or more intermediates. Recently, Li and co-workers found a linear correlation between the unfolding force of the native state and the intermediate in fibronectin, which they suggested indicated the presence of a molecular memory or multiple unfolding pathways (1). Here, we apply two independent methods in combination with Monte Carlo simulations to analyze the unfolding of alpha-helices E and D of bacteriorhodopsin (BR). We show that correlation analysis of unfolding forces is very sensitive to errors in force calibration of the instrument. In contrast, a comparison of relative forces provides a robust measure for the stability of unfolding intermediates. The proposed approach detects three energetically different states of alpha-helices E and D in trimeric BR. These states are not observed for monomeric BR and indicate that substantial information is hidden in forced unfolding experiments of single proteins. PMID- 15792968 TI - Coupling field theory with continuum mechanics: a simulation of domain formation in giant unilamellar vesicles. AB - Domain formation is modeled on the surface of giant unilamellar vesicles using a Landau field theory model for phase coexistence coupled to elastic deformation mechanics (e.g., membrane curvature). Smooth particle applied mechanics, a form of smoothed particle continuum mechanics, is used to solve either the time dependent Landau-Ginzburg or Cahn-Hilliard free-energy models for the composition dynamics. At the same time, the underlying elastic membrane is modeled using smooth particle applied mechanics, resulting in a unified computational scheme capable of treating the response of the composition fields to arbitrary deformations of the vesicle and vice versa. The results indicate that curvature coupling, along with the field theory model for composition free energy, gives domain formations that are correlated with surface defects on the vesicle. In the case that external deformations are included, the domain structures are seen to respond to such deformations. The present simulation capability provides a significant step forward toward the simulation of realistic cellular membrane processes. PMID- 15792969 TI - A consistent experimental and modeling approach to light-scattering studies of protein-protein interactions in solution. AB - The osmotic second virial coefficient, B(2), obtained by light scattering from protein solutions has two principal components: the Donnan contribution and a contribution due to protein-protein interactions in the limit of infinite dilution. The Donnan contribution accounts for electroneutrality in a multicomponent solution of (poly)electrolytes. The importance of distinguishing this ideal contribution to B(2) is emphasized, thereby allowing us to model the interaction part of B(2) by molecular computations. The model for protein-protein interactions that we use here extends earlier work (Neal et al., 1998) by accounting for long-range electrostatic interactions and the specific hydration of the protein by strongly associated water molecules. Our model predictions are compared with measurements of B(2) for lysozyme at 25 degrees C over pH from 5.0 to 9.0, and 7-60 mM ionic strength. We find that B(2) is positive at all solution conditions and decreases with increasing ionic strength, as expected, whereas the interaction part of B(2) is negative at all conditions and becomes progressively less negative with increasing ionic strength. Although long-range electrostatic interactions dominate this contribution, particularly at low ionic strength, short-range electrostatic/dispersion interactions with specific hydration are essential for an accurate description of B(2) derived from experiment. PMID- 15792971 TI - Fluctuation correlation spectroscopy with a laser-scanning microscope: exploiting the hidden time structure. AB - Images obtained with a laser-scanning microscope contain a time structure that can be exploited to measure fast dynamics of molecules in solution and in cells. The spatial correlation approach provides a simple algorithm to extract this information. We describe the analysis used to process laser-scanning images of solutions and cells to obtain molecular diffusion constant in the microsecond to second timescale. PMID- 15792970 TI - Two-photon cross-correlation analysis of intracellular reactions with variable stoichiometry. AB - We successfully demonstrate the effectiveness of two-photon fluorescence cross correlation spectroscopy (TPCCS) to study the complex binding stoichiometry of calmodulin (CaM) and Ca(2+)/CaM-dependent protein kinase II (CaMKII). Practical considerations are made for developing an intracellular cross-correlation assay, including characterization of the fluorescent molecules involved, calibration procedures of the setup, and optimal measurement conditions. Potential pitfalls and artifacts are discussed, and the complex stoichiometry of the molecular system is accounted for by a new experimental and theoretical framework for TPCCS. Our tailored model accommodates up to 12 red-labeled CaMs binding to a single green-labeled dodecameric CaMKII holoenzyme and accounts for the probability distributions of bound ligand as well as the respective changes in fluorescence emission upon binding. The model was experimentally demonstrated both in solution and in living cells by analyzing the binding of Alexa 633(C2)CaM to eGFP-CaMKII under different biochemical conditions known to induce the basal, activated, and autophosphorylated forms of the enzyme. Key binding parameters, such as binding degree, concentrations of reactants, and binding affinities, were determined under varying conditions with certain assumptions. TPCCS thus offers the unique ability to test our biochemical understanding of protein dynamics in the intracellular milieu. PMID- 15792972 TI - Thermodynamic stability of a kappaI immunoglobulin light chain: relevance to multiple myeloma. AB - Immunoglobulin light chains have two similar domains, each with a hydrophobic core surrounded by beta-sheet layers, and a highly conserved disulfide bond. Differential scanning calorimetry and circular dichroism were used to study the folding and stability of MM-kappaI, an Ig LC of kappaI subtype purified from the urine of a multiple myeloma patient. The complete primary structure of MM-kappaI was determined by Edman sequence analysis and mass spectrometry. The protein was found to contain a cysteinyl post-translational modification at Cys(214). Protein stability and conformation of MM-kappaI as a function of temperature or denaturant conditions at pH 7.4 and 4.8 were investigated. At pH 4.8, calorimetry demonstrated that MM-kappaI undergoes an incomplete, cooperative, partially reversible thermal unfolding with increased unfolding temperature and calorimetric enthalpy as compared to pH 7.4. Secondary and tertiary structural analyses provided evidence to support the presence of unfolding intermediates. Chemical denaturation resulted in more extensive protein unfolding. The stability of MM-kappaI was reduced and protein unfolding was irreversible at pH 4.8, thus suggesting that different pathways are utilized in thermal and chemical unfolding. PMID- 15792973 TI - Oligomerization endows enormous stability to soybean agglutinin: a comparison of the stability of monomer and tetramer of soybean agglutinin. AB - Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. This protein shows a very high degree of stability when compared to the other proteins of the same family. In a previous work, it was shown that the unusual stability of the protein is due to a high degree of subunit interactions. In this study we present the thermodynamic parameters for the stability of soybean agglutinin monomer. The monomeric species is found at pH 2 and below which it is most populated at pH 1.9, as evident from size-exclusion chromatographic and dynamic light scattering studies. The analyses of circular dichroism and fluorescence spectroscopy suggest that the monomer is well folded, and that it has certain characteristic features when compared to its tetrameric counterpart. The conformational stabilities of the tetramer and the monomer at the temperature of their maximum stabilities (310 K) are 59.2 kcal/mol and 9.8 kcal/mol, respectively, indicating that oligomerization contributes significantly to the stability of the native molecule. Also, the T(g) difference for the two forms of the protein is approximately 40 K, whereas the difference in DeltaC(p) is only 1.6 kcal/mol/K. This suggests that the major hydrophobic core is present in the monomer itself, and that oligomerization involves mainly ionic interactions. PMID- 15792974 TI - Ionic determinants of functional reentry in a 2-D model of human atrial cells during simulated chronic atrial fibrillation. AB - Recent studies suggest that atrial fibrillation (AF) is maintained by fibrillatory conduction emanating from a small number of high-frequency reentrant sources (rotors). Our goal was to study the ionic correlates of a rotor during simulated chronic AF conditions. We utilized a two-dimensional (2-D), homogeneous, isotropic sheet (5 x 5 cm(2)) of human atrial cells to create a chronic AF substrate, which was able to sustain a stable rotor (dominant frequency approximately 5.7 Hz, rosette-like tip meander approximately 2.6 cm). Doubling the magnitude of the inward rectifier K(+) current (I(K1)) increased rotor frequency ( approximately 8.4 Hz), and reduced tip meander (approximately 1.7 cm). This rotor stabilization was due to a shortening of the action potential duration and an enhanced cardiac excitability. The latter was caused by a hyperpolarization of the diastolic membrane potential, which increased the availability of the Na(+) current (I(Na)). The rotor was terminated by reducing the maximum conductance (by 90%) of the atrial-specific ultrarapid delayed rectifier K(+) current (I(Kur)), or the transient outward K(+) current (I(to)), but not the fast or slow delayed rectifier K(+) currents (I(Kr)/I(Ks)). Importantly, blockade of I(Kur)/I(to) prolonged the atrial action potential at the plateau, but not at the terminal phase of repolarization, which led to random tip meander and wavebreak, resulting in rotor termination. Altering the rectification profile of I(K1) also slowed down or abolished reentrant activity. In combination, these simulation results provide novel insights into the ionic bases of a sustained rotor in a 2-D chronic AF substrate. PMID- 15792975 TI - Effect of cell electroporation on the conductivity of a cell suspension. AB - An increased permeability of a cell membrane during the application of high voltage pulses results in increased transmembrane transport of molecules that otherwise cannot enter the cell. Increased permeability of a cell membrane is accompanied by increased membrane conductivity; thus, by measuring electric conductivity the extent of permeabilized tissue could be monitored in real time. In this article the effect of cell electroporation caused by high-voltage pulses on the conductivity of a cell suspension was studied by current-voltage measurements during and impedance measurement before and after the pulse application. At the same time the percentage of permeabilized and survived cells was determined and the extent of osmotic swelling measured. For a train of eight pulses a transient increase in conductivity of a cell suspension was obtained above permeabilization threshold in low- and high-conductive medium with complete relaxation in <1 s. Total conductivity changes and impedance measurements showed substantial changes in conductivity due to the ion efflux in low-conductive medium and colloid-osmotic swelling in both media. Our results show that by measuring electric conductivity during the pulses we can detect limit permeabilization threshold but not directly permeabilization level, whereas impedance measurements in seconds after the pulse application are not suitable. PMID- 15792976 TI - InsP3 signaling induces pulse-modulated Ca2+ signals in the nucleus of airway epithelial ciliated cells. AB - The phenomenology of nuclear Ca(2+) dynamics has experienced important progress revealing the broad range of cellular processes that it regulates. Although several agonists can mobilize Ca(2+) from storage in the nuclear envelope (NE) to the intranuclear compartment (INC), the mechanisms of Ca(2+) signaling in the nucleus still remain uncertain. Here we report that the NE/INC complex can function as an inositol-1,4,5-trisphosphate (InsP(3))-controlled Ca(2+) oscillator. Thin optical sectioning combined with fluorescent labeling of Ca(2+) probes show in cultured airway epithelial ciliated cells that ATP can trigger periodic oscillations of Ca(2+) in the NE ([Ca(2+)](NE)) and corresponding pulses of Ca(2+) release to the INC. Identical results were obtained in InsP(3) stimulated isolated nuclei of these cells. Our data show that [Ca(2+)](NE) oscillations and Ca(2+) release to the INC result from the interplay between the Ca(2+)/K(+) ion-exchange properties of the intralumenal polyanionic matrix of the NE and two Ca(2+)-sensitive ion channels-an InsP(3)-receptor-Ca(2+) channel and an apamin-sensitive K(+) channel. A similar Ca(2+) signaling system operating under the same functional protocol and molecular hardware controls Ca(2+) oscillations and release in/to the endoplasmic reticulum/cytosol and in/to the granule/cytosol complexes in airway and mast cells. These observations suggest that these intracellular organelles share a remarkably conserved mechanism of InsP(3)-controlled frequency-encoded Ca(2+) signaling. PMID- 15792977 TI - Elastic lever-arm model for myosin V. AB - We present a mechanochemical model for myosin V, a two-headed processive motor protein. We derive the properties of a dimer from those of an individual head, which we model both with a four-state cycle (detached; attached with ADP.Pi; attached with ADP; and attached without nucleotide) and alternatively with a five state cycle (where the powerstroke is not tightly coupled to the phosphate release). In each state the lever arm leaves the head at a different, but fixed, angle. The lever arm itself is described as an elastic rod. The chemical cycles of both heads are coordinated exclusively by the mechanical connection between the two lever arms. The model explains head coordination by showing that the lead head only binds to actin after the powerstroke in the trail head and that it only undergoes its powerstroke after the trail head unbinds from actin. Both models (four- and five-state) reproduce the observed hand-over-hand motion and fit the measured force-velocity relations. The main difference between the two models concerns the load dependence of the run length, which is much weaker in the five state model. We show how systematic processivity measurement under varying conditions could be used to distinguish between both models and to determine the kinetic parameters. PMID- 15792979 TI - Role of the endothelium on arterial vasomotion. AB - It is well-known that cyclic variations of the vascular diameter, a phenomenon called vasomotion, are induced by synchronous calcium oscillations of smooth muscle cells (SMCs). However, the role of the endothelium on vasomotion is unclear. Some experimental studies claim that the endothelium is necessary for synchronization and vasomotion, whereas others report rhythmic contractions in the absence of an intact endothelium. Moreover, endothelium-derived factors have been shown to abolish vasomotion by desynchronizing the calcium signals in SMCs. By modeling the calcium dynamics of a population of SMCs coupled to a population of endothelial cells, we analyze the effects of an SMC vasoconstrictor stimulation on endothelial cells and the feedback of endothelium-derived factors. Our results show that the endothelium essentially decreases the SMCs calcium level and may move the SMCs from a steady state to an oscillatory domain, and vice versa. In the oscillatory domain, a population of coupled SMCs exhibits synchronous calcium oscillations. Outside the oscillatory domain, the coupled SMCs present only irregular calcium flashings arising from noise modeling stochastic opening of channels. Our findings provide explanations for the published contradictory experimental observations. PMID- 15792978 TI - Effects of sucrose on the internal dynamics of azurin. AB - Sucrose is a natural osmolyte accumulated in cells of organisms as they adapt to environmental stresses. In vitro, sucrose increases protein stability and forces partially unfolded structures to refold. Its effects on the native fold structure and dynamics are not fully established. This study, utilizing Trp phosphorescence spectroscopy, examined the influence of molar concentrations of sucrose on the flexibility of metal-free azurin from Pseudomonas aeruginosa. In addition, by means of specific mutants of the test protein, namely I7S, F110S, and C3A/C26A, that altered its thermodynamic stability, its intrinsic flexibility, and the extent of internal hydration, this investigation sought to identify possible correlations between these features of protein structure and the influence of the osmolyte on protein dynamics. Alterations of structural fluctuations were assessed by both the intrinsic phosphorescence lifetime (tau), which reports on local structure about the triplet probe, and the acrylamide bimolecular quenching rate constant (k(q)) that is a measure of the average acrylamide diffusion coefficient through the macromolecule. From the modulation of tau and k(q) across a wide temperature range and up to a concentration of 2M sucrose, it is concluded that sucrose attenuates structural fluctuations principally when macromolecules are internally hydrated and thermally expanded. Preliminary tests with trehalose and xylitol suggest that the effects of sucrose are general of the polyol class of osmolytes. PMID- 15792980 TI - Persistence length of titin from rabbit skeletal muscles measured with scattering and microrheology techniques. AB - The persistence length of titin from rabbit skeletal muscles was measured using a combination of static and dynamic light scattering, and neutron small angle scattering. Values of persistence length in the range 9-16 nm were found for titin-II, which corresponds to mainly physiologically inelastic A-band part of the protein, and for a proteolytic fragment with 100-nm contour length from the physiologically elastic I-band part. The ratio of the hydrodynamic radius to the static radius of gyration indicates that the proteins obey Gaussian statistics typical of a flexible polymer in a -solvent. Furthermore, measurements of the flexibility as a function of temperature demonstrate that titin-II and the I-band titin fragment experience a similar denaturation process; unfolding begins at 318 K and proceeds in two stages: an initial gradual 50% change in persistence length is followed by a sharp unwinding transition at 338 K. Complementary microrheology (video particle tracking) measurements indicate that the viscoelasticity in dilute solution behaves according to the Flory/Fox model, providing a value of the radius of gyration for titin-II (63 +/- 1 nm) in agreement with static light scattering and small angle neutron scattering results. PMID- 15792982 TI - Kinetics and thermodynamics of association of a fluorescent lysophospholipid derivative with lipid bilayers in liquid-ordered and liquid-disordered phases. AB - We have measured the rates of insertion into, desorption from, and spontaneous interlayer translocation (flip-flop) of the fluorescent lysophospholipid derivative NBD-lyso-1-myristoylphosphatidylethanolamine in l(d) and l(o) phase lipid bilayer membranes. The lipid bilayers, studied as LUV, were prepared from pure 1-palmitoyl-2-oleoylphosphatidylcholine, in the l(d) phase; and from two Chol-containing binary lipid mixtures, 1-palmitoyl-2-oleoylphosphatidylcholine and Chol (molar ratio of 1:1) and SpM and Chol (molar ratio of 6:4), both in the l(o) phase. Insertion, desorption, and translocation rate constants and equilibrium constants for association of the amphiphile monomer with the lipid bilayers were measured between 15 degrees C and 35 degrees C, and the standard free energies, enthalpies, and entropies, as well as the activation energies for these processes were derived from these data. The equilibrium partition coefficients for partitioning of the amphiphile between the aqueous phase and the different membrane phases were also derived, and an estimation was made of hypothetical partition coefficients and the respective energetic parameters for partitioning between the different lipid phases if these were to coexist in the same membrane. We show that, contrary to general belief, the association of NBD lysoMPE with lipid bilayers is not a diffusion-controlled process, the rate limiting step in insertion being the formation of a free area in the membrane surface of an adequate size for insertion to occur. PMID- 15792981 TI - Domain formation and stability in complex lipid bilayers as reported by cholestatrienol. AB - In this study, we used cholestatrienol (CTL) as a fluorescent reporter molecule to study sterol-rich L(o) domains in complex lipid bilayers. CTL is a fluorescent cholesterol analog that mimics the behavior of cholesterol well. The ability of 12SLPC to quench the fluorescence of cholestatrienol gives a measure of the amount of sterol included in L(o) domains in mixed lipid membranes. The stability of sterol-rich domains formed in complex lipid mixtures containing saturated sphingomyelins, phosphatidylcholines, or galactosylceramide as potential domain forming lipids were studied. The amount of sterol associated with sterol-rich domains seemed to always increase with increasing temperature. The quenching efficiency was highly dependent on the domain-forming lipid present in complex lipid mixtures. Sphingomyelins formed stable sterol-enriched domains and were able to shield CTL from quenching better than the other lipids included in this study. The saturated phosphatidylcholines also formed sterol-rich domains, but the quenching efficiency in membranes with these was higher than with sphingomyelins and the domains melted at lower temperatures. PGalCer was not able to form sterol-enriched domains. However, we found that PGalCer stabilized sterol rich domains formed in PSM-containing bilayers. Using a fluorescent ceramide analog, we also demonstrated that N-palmitoyl-ceramide displaced the sterol from sphingolipid-rich domains in mixed bilayer membranes. PMID- 15792983 TI - Correlation between vesicle quantal size and fusion pore release in chromaffin cell exocytosis. AB - A significant number of exocytosis events recorded with amperometry demonstrate a prespike feature termed a "foot" and this foot has been correlated with messengers released via a transitory fusion pore before full exocytosis. We have compared amperometric spikes with a foot with spikes without a foot at chromaffin cells and found that the probability of detecting a distinct foot event is correlated to the amount of catecholamine released. The mean charge of the spikes with a foot was found to be twice that of the spikes without a foot, and the frequency of spikes displaying a foot was zero for small spikes increasing to approximately 50% for large spikes. It is hypothesized that in chromaffin cells, where the dense core is believed to nearly fill the vesicle, the expanding core is a controlling factor in opening the fusion pore, that prefusion of two smaller vesicles leads to excess membrane, and that this slows pore expansion leading to an increased observation of events with a foot. Clearly, the physicochemical properties of vesicles are key factors in the control of the dynamics of release through the fusion pore and the high and variable frequency of this release makes it highly significant. PMID- 15792984 TI - Ultrafast excited state dynamics of the protonated Schiff base of all-trans retinal in solvents. AB - We present a comparative study of the ultrafast photophysics of all-trans retinal in the protonated Schiff base form in solvents with different polarities and viscosities. Steady-state spectra of retinal in the protonated Schiff base form show large absorption-emission Stokes shifts (6500-8100 cm(-1)) for both polar and nonpolar solvents. Using a broadband fluorescence up-conversion experiment, the relaxation kinetics of fluorescence is investigated with 120 fs time resolution. The time-zero spectra already exhibit a Stokes-shift of approximately 6000 cm(-1), indicating depopulation of the Franck-Condon region in < or =100 fs. We attribute it to relaxation along skeletal stretching. A dramatic spectral narrowing is observed on a 150 fs timescale, which we assign to relaxation from the S(2) to the S(1) state. Along with the direct excitation of S(1), this relaxation populates different quasistationary states in S(1), as suggested from the existence of three distinct fluorescence decay times with different decay associated spectra. A 0.5-0.65 ps decay component is observed, which may reflect the direct repopulation of the ground state, in line with the small isomerization yield in solvents. Two longer decay components are observed and are attributed to torsional motion leading to photo-isomerization. The various decay channels show little or no dependence with respect to the viscosity or dielectric constant of the solvents. This suggests that in the protein, the bond selectivity of isomerization is mainly governed by steric effects. PMID- 15792985 TI - Chronic inflammatory pain leads to increased blood-brain barrier permeability and tight junction protein alterations. AB - The blood-brain barrier (BBB) maintains brain homeostasis by limiting entry of substances to the central nervous system through interaction of transmembrane and intracellular proteins that make up endothelial cell tight junctions (TJs). Recently it was shown that the BBB can be modulated by disease pathologies including inflammatory pain. This study examined the effects of chronic inflammatory pain on the functional and molecular integrity of the BBB. Inflammatory pain was induced by injection of complete Freund's adjuvant (CFA) into the right plantar hindpaw in female Sprague-Dawley rats under halothane anesthesia; control animals were injected with saline. Edema and hyperalgesia were assessed by plethysmography and infrared paw-withdrawal latency. At 72 h postinjection, significant edema formation and hyperalgesia were noted in the CFA treated rats. Examination of permeability of the BBB by in situ perfusion of [14C]sucrose while rats were under pentobarbital anesthesia demonstrated that CFA treatment significantly increased brain sucrose uptake. Western blot analysis of BBB TJ proteins showed no change in expression of zonula occludens-1 (an accessory protein) or actin (a cytoskeletal protein) with CFA treatment. Expression of the transmembrane TJ proteins occludin and claudin-3 and -5 significantly changed with CFA treatment with a 60% decrease in occludin, a 450% increase in claudin-3, and a 615% increase in claudin-5 expression. This study demonstrates that during chronic inflammatory pain, alterations in BBB function are associated with changes in specific transmembrane TJ proteins. PMID- 15792986 TI - Moderate endurance training prevents doxorubicin-induced in vivo mitochondriopathy and reduces the development of cardiac apoptosis. AB - The objective of this work was to test the hypothesis that endurance training may be protective against in vivo doxorubicin (DOX)-induced cardiomyopathy through mitochondria-mediated mechanisms. Forty adult (6-8 wk old) male Wistar rats were randomly divided into four groups (n = 10/group): nontrained, nontrained + DOX treatment (20 mg/kg), trained (14 wk of endurance treadmill running, 60-90 min/day), and trained + DOX treatment. Mitochondrial respiration, calcium tolerance, oxidative damage, heat shock proteins (HSPs), antioxidant enzyme activity, and apoptosis markers were evaluated. DOX induces mitochondrial respiratory dysfunction, oxidative damage, and histopathological lesions and triggers apoptosis (P < 0.05, n = 10). However, training limited the decrease in state 3 respiration, respiratory control ratio (RCR), uncoupled respiration, aconitase activity, and protein sulfhydryl content caused by DOX treatment and prevented the increased sensitivity to calcium in nontrained + DOX-treated rats (P < 0.05, n = 10). Moreover, training inhibited the DOX-induced increase in mitochondrial protein carbonyl groups, malondialdehyde, Bax, Bax-to-Bcl-2 ratio, and tissue caspase-3 activity (P < 0.05, n = 10). Training also increased by approximately 2-fold the expression of mitochondrial HSP-60 and tissue HSP-70 (P < 0.05, n = 10) and by approximately 1.5-fold the activity of mitochondrial and cytosolic forms of SOD (P < 0.05, n = 10). We conclude that endurance training protects heart mitochondrial respiratory function from the toxic effects of DOX, probably by improving mitochondrial and cell defense systems and reducing cell oxidative stress. In addition, endurance training limited the DOX-triggered apoptosis. PMID- 15792987 TI - Power output is linearly related to MyHC content in rat skinned myocytes and isolated working hearts. AB - The amount of work the heart can perform during ejection is governed by the inherent contractile properties of individual myocytes. One way to alter contractile properties is to alter contractile proteins such as myosin heavy chain (MyHC), which is known to demonstrate isoform plasticity in response to disease states. The purpose of this study was to examine myocyte functionality over the complete range of MyHC expression in heart, from 100% alpha-MyHC to 100% beta-MyHC, using euthyroid and hypothyroid rats. Peak power output in skinned cardiac myocytes decreased as a nearly linear function of beta-MyHC expression during maximal (r2 = 0.85, n = 44 myocyte preparations) and submaximal (r2 = 0.82, n = 31 myocyte preparations) Ca2+ activation. To determine whether single myocyte function translated to the level of the whole heart, power output was measured in working heart preparations expressing varied ratios of MyHC. Left ventricular power output of isolated working heart preparations also decreased as a linear function of increasing beta-MyHC expression (r2 = 0.82, n = 34 myocyte preparations). These results demonstrate that power output is highly dependent on MyHC expression in single myocytes, and this translates to the performance of working left ventricles. PMID- 15792988 TI - RV filling modulates LV function by direct ventricular interaction during mechanical ventilation. AB - During mechanical ventilation, phasic changes in systemic venous return modulate right ventricular output but may also affect left ventricular function by direct ventricular interaction. In 13 anesthetized, closed-chest, normal dogs, we measured inferior vena cava flow and left and right ventricular dimensions and output during mechanical ventilation, during an inspiratory hold, and (during apnea) vena caval constriction and abdominal compression. During a single ventilation cycle preceded by apnea, positive pressure inspiration decreased caval flow and right ventricular dimension; the transseptal pressure gradient increased, the septum shifted rightward, reflecting an increased left ventricular volume (the anteroposterior diameter did not change); and stroke volume increased. The opposite occurred during expiration. Similarly, the maneuvers that decreased venous return shifted the septum rightward, and left ventricular volume and stroke volume increased. Increased venous return had opposite effects. Changes in left ventricular function caused by changes in venous return alone were similar to those during mechanical ventilation except for minor quantitative differences. We conclude that phasic changes in systemic venous return during mechanical ventilation modulate left ventricular function by direct ventricular interaction. PMID- 15792989 TI - Mechanisms of superiority of ascending ramp waveforms: new insights into mechanisms of shock-induced vulnerability and defibrillation. AB - Monophasic ascending ramp (AR) and descending ramp (DR) waveforms are known to have significantly different defibrillation thresholds. We hypothesized that this difference arises due to differences in mechanisms of arrhythmia induction for the two waveforms. Rabbit hearts (n = 10) were Langendorff perfused, and AR and DR waveforms (7, 20, and 40 ms) were randomly delivered from two line electrodes placed 10 mm apart on the anterior ventricular epicardium. We optically mapped cellular responses to shocks of various strengths (5, 10, and 20 V/cm) and coupling intervals (CIs; 120, 180, and 300 ms). Optical mapping revealed that maximum virtual electrode polarization (VEP) was reached at significantly different times for AR and DR of the same duration (P < 0.05) for all tested CIs. As a result, VEP for AR were stronger than for DR at the end of the shock. Postshock break excitation resulting from AR generated faster propagation and typically could not form reentry. In contrast, partially dissipated VEP resulting from DR generated slower propagation; the wavefront was able to propagate into deexcited tissue and thus formed a shock-induced reentry circuit. Therefore, for the same delivered energy, AR was less proarrhythmic compared with DR. An active bidomain model was used to confirm the electrophysiological results. The VEP hypothesis explains differences in vulnerability associated with monophasic AR and DR waveforms and, by extension, the superior defibrillation efficacy of the AR waveform compared with the DR waveform. PMID- 15792990 TI - Reduced molecular expression of K(+) channel proteins in vascular smooth muscle from rats made hypertensive with N{omega}-nitro-L-arginine. AB - Smooth muscle membrane potential (E(m)) depends on K(+) channels, and arteries from rats made hypertensive with N(omega)-nitro-l-arginine (LHR) are depolarized compared with control. We hypothesized that decreased K(+) channel function, due to decreased K(+) channel protein expression, underlies E(m) depolarization. Furthermore, K(+) channel blockers should move control E(m) (-46 +/- 1 mV) toward that in LHR (-37 +/- 2 mV) and normalize contraction. The E(m) vs. K(+) relationship was less steep in LHR (23 +/- 2 vs. 28 +/- 1 mV/log K(+) concentration), and contractile sensitivity to K(+) was increased (EC(50) = 37 +/ 1 vs. 23 +/- 1 mM). Iberiotoxin (10 nM), an inhibitor of large-conductance Ca(2+)-activated K(+) (BK(Ca)) channels, depolarized control and LHR E(m) to -35 +/- 1 and -30 +/- 2 mV, respectively; however, effects on K(+) sensitivity were more profound in LHR (EC(50) = 25 +/- 2 vs. 15 +/- 3 mM). The voltage-dependent K(+) (K(V)) channel blocker 4-aminopyridine (3 mM) depolarized control E(m) to the level of LHR (-28 +/- 1 vs. -28 +/- 1 mV); however, effects on K(+) sensitivity were greater in LHR (EC(50) = 17 +/- 4 vs. 4 +/- 4 mM). Western blots revealed reduced BK(Ca) and K(V)1.5 channel expression in LHR arteries. The findings suggest that diminished expression of K(+) channels contributes to depolarization and enhanced contractile sensitivity. These conclusions are supported by direct electrophysiological assessment of BK(Ca) and K(V) channel function in control and LHR smooth muscle cells. PMID- 15792992 TI - Analysis of blood flow in the entire coronary arterial tree. AB - A hemodynamic analysis of coronary blood flow must be based on the measured branching pattern and vascular geometry of the coronary vasculature. We recently developed a computer reconstruction of the entire coronary arterial tree of the porcine heart based on previously measured morphometric data. In the present study, we carried out an analysis of blood flow distribution through a network of millions of vessels that includes the entire coronary arterial tree down to the first capillary branch. The pressure and flow are computed throughout the coronary arterial tree based on conservation of mass and momentum and appropriate pressure boundary conditions. We found a power law relationship between the diameter and flow of each vessel branch. The exponent is approximately 2.2, which deviates from Murray's prediction of 3.0. Furthermore, we found the total arterial equivalent resistance to be 0.93, 0.77, and 1.28 mmHg.ml(-1).s(-1).g(-1) for the right coronary artery, left anterior descending coronary artery, and left circumflex artery, respectively. The significance of the present study is that it yields a predictive model that incorporates some of the factors controlling coronary blood flow. The model of normal hearts will serve as a physiological reference state. Pathological states can then be studied in relation to changes in model parameters that alter coronary perfusion. PMID- 15792991 TI - Protein kinase C-zeta inhibition exerts cardioprotective effects in ischemia reperfusion injury. AB - Ischemia followed by reperfusion (I/R) in the presence of polymorphonuclear leukocytes (PMNs) results in marked cardiac contractile dysfunction. A cell permeable PKC-zeta peptide inhibitor was used to test the hypothesis that PKC zeta inhibition could attenuate PMN-induced cardiac contractile dysfunction by suppression of superoxide production from PMNs and increase nitric oxide (NO) release from vascular endothelium. The effects of the PKC-zeta peptide inhibitor were examined in isolated ischemic (20 min) and reperfused (45 min) rat hearts reperfused with PMNs. The PKC-zeta inhibitor (2.5 or 5 microM, n = 6) significantly attenuated PMN-induced cardiac dysfunction compared with I/R hearts (n = 6) receiving PMNs alone in left ventricular developed pressure (LVDP) and the maximal rate of LVDP (+dP/dt(max)) cardiac function indexes (P < 0.01), and these cardioprotective effects were blocked by the NO synthase inhibitor, N(G) nitro-L-arginine methyl ester (50 microM). Furthermore, the PKC-zeta inhibitor significantly increased endothelial NO release 47 +/- 2% (2.5 microM, P < 0.05) and 54 +/- 5% (5 microM, P < 0.01) over basal values from the rat aorta and significantly inhibited superoxide release from phorbol-12-myristate-13-acetate stimulated rat PMNs by 33 +/- 12% (2.5 microM) and 40 +/- 8% (5 microM) (P < 0.01). The PKC-zeta inhibitor significantly attenuated PMN infiltration into the myocardium by 46-48 +/- 4% (P < 0.01) at 2.5 and 5 microM, respectively. In conclusion, these results suggest that the PKC-zeta peptide inhibitor attenuates PMN-induced post-I/R cardiac contractile dysfunction by increasing endothelial NO release and by inhibiting superoxide release from PMNs thereby attenuating PMN infiltration into I/R myocardium. PMID- 15792993 TI - Biaxial elastic material properties of porcine coronary media and adventitia. AB - The importance of mechanical stresses and strains has become well recognized in vascular physiology and pathology. To compute the stress and strain on the various components of the vessel wall, we must know the constitutive equations for the different layers of the vessel wall. The objective of the present study is to determine the constitutive equation of the coronary artery treated as a two layer composite: intima-media and adventitial layers. Twelve hearts were obtained from a local slaughterhouse, and the right coronary artery and left anterior descending artery were dissected free from the myocardium. The vessel wall was initially mechanically tested biaxially (inflation and axial extension) as a whole (intact wall) and subsequently as intima-media or adventitial layer. A Fung type exponential strain energy function was used to curve fit the experimental data for the intact wall and individual layers for the right coronary artery and left anterior descending artery. Two methods were used for the determination of material constants, including the Marquardt-Levenberg nonlinear least squares method and the genetic algorithm method. Our results show that there were no statistically significant differences in the material constants obtained from the two methods and that either set of elastic constants results in good fit of the data. Furthermore, at an in vivo value of axial stretch ratio, we find that the stiffness is as follows: intima-media > intact > adventitia. These results underscore the composite nature of coronary arteries with different material properties in each layer. The present results are necessary for analysis of coronary artery mechanics and to provide a fundamental understanding of vessel physiology. PMID- 15792994 TI - Insulin resistance in spontaneously hypertensive rats is associated with endothelial dysfunction characterized by imbalance between NO and ET-1 production. AB - Insulin stimulates production of NO in vascular endothelium via activation of phosphatidylinositol (PI) 3-kinase, Akt, and endothelial NO synthase. We hypothesized that insulin resistance may cause imbalance between endothelial vasodilators and vasoconstrictors (e.g., NO and ET-1), leading to hypertension. Twelve-week-old male spontaneously hypertensive rats (SHR) were hypertensive and insulin resistant compared with control Wistar-Kyoto (WKY) rats (systolic blood pressure 202 +/- 11 vs. 132 +/- 10 mmHg; fasting plasma insulin 5 +/- 1 vs. 0.9 +/- 0.1 ng/ml; P < 0.001). In WKY rats, insulin stimulated dose-dependent relaxation of mesenteric arteries precontracted with norepinephrine (NE) ex vivo. This depended on intact endothelium and was blocked by genistein, wortmannin, or N(omega)-nitro-l-arginine methyl ester (inhibitors of tyrosine kinase, PI3 kinase, and NO synthases, respectively). Vasodilation in response to insulin (but not ACh) was impaired by 20% in SHR (vs. WKY, P < 0.005). Preincubation of arteries with insulin significantly reduced the contractile effect of NE by 20% in WKY but not SHR rats. In SHR, the effect of insulin to reduce NE-mediated vasoconstriction became evident when insulin pretreatment was accompanied by ET-1 receptor blockade (BQ-123, BQ-788). Similar results were observed during treatment with the MEK inhibitor PD-98059. In addition, insulin-stimulated secretion of ET-1 from primary endothelial cells was significantly reduced by pretreatment of cells with PD-98059 (but not wortmannin). We conclude that insulin resistance in SHR is accompanied by endothelial dysfunction in mesenteric vessels with impaired PI3-kinase-dependent NO production and enhanced MAPK dependent ET-1 secretion. These results may reflect pathophysiology in other vascular beds that directly contribute to elevated peripheral vascular resistance and hypertension. PMID- 15792995 TI - Inhibition of platelet functions and thrombosis through selective or nonselective inhibition of the platelet P2 receptors with increasing doses of NF449 [4,4',4'',4'''-(carbonylbis(imino-5,1,3-benzenetriylbis-(carbonylimino)))tetrakis benzene-1,3-disulfonic acid octasodium salt]. AB - Our aim was to determine whether the newly described P2X1 antagonist NF449 [4,4',4'',4'''-(carbonylbis(imino-5,1,3-benzenetriylbis(carbonylimino)))tetrakis benzene-1,3-disulfonic acid octasodium salt] could selectively antagonize the platelet P2X1 receptor and how it affected platelet function. NF449 inhibited alpha,beta-methyleneadenosine 5'-triphosphate-induced shape change (IC50 = 83 +/- 13 nM; n = 3) and calcium influx (pA2 = 7.2 +/- 0.1; n = 3) (pIC50 = 6.95) in washed human platelets treated with apyrase to prevent desensitization of the P2X1 receptor. NF449 also antagonized the calcium rise mediated by the P2Y1 receptor, but with lower potency (IC50 = 5.8 +/- 2.2 microM; n = 3). In contrast, it was a very weak antagonist of the P2Y12-mediated inhibition of adenylyl cyclase activity. Selective blockade of the P2X1 receptor with NF449 led to reduced collagen-induced aggregation, confirming a role of this receptor in platelet activation induced by collagen. Intravenous injection of 10 mg/kg NF449 into mice resulted in selective inhibition of the P2X1 receptor and decreased intravascular platelet aggregation in a model of systemic thromboembolism (35 +/- 4 versus 51 +/- 3%) (P = 0.0061; n = 10) but without prolongation of the bleeding time (106 +/- 16 versus 78 +/- 7 s; n = 10) (N.S.; P = 0.1209). At a higher dose (50 mg/kg), NF449 inhibited the three platelet P2 receptors. This led to a further reduction in platelet consumption compared with mice injected with saline (13 +/- 4 versus 42 +/- 3%) (P = 0.0002; n = 5). NF449 also reduced dose dependently the size of thrombi formed after laser-induced injury of mesenteric arterioles. Overall, our results indicate that NF449 constitutes a new tool to investigate the functions of the P2X1 receptor and could be a starting compound in the search for new antithrombotic drugs targeting the platelet P2 receptors. PMID- 15792996 TI - BAY 41-2272 [5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3 yl]pyrimidin-4-ylamine]-induced dilation in ovine pulmonary artery: role of sodium pump. AB - The mechanisms of relaxation to nitric oxide (NO)-independent soluble guanylyl cyclase (sGC) activator BAY 41-2272 [5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H pyrazolo[3,4-b]pyridine-3-yl]pyrimidin-4-ylamine] were investigated in isolated ovine pulmonary artery. BAY 41-2272 (1 nM-10 microM) produced concentration dependent relaxation of endothelium-denuded pulmonary artery rings (pD2 = 6.82 +/ 0.16; Emax = 92.30 +/- 2.31%; n = 8), precontracted with 1 microM 5 hydroxytryptamine (serotonin). 1-H-[1,2,4]Oxadiazole[4,3-a]quinoxalin-1-one (ODQ; 10 microM), an inhibitor of sGC, partially inhibited (Emax = 57.10 +/- 3.10%; n = 6) the relaxation response of BAY 41-2272. In comparison with ODQ, sodium pump inhibitor ouabain (1 microM) produced a greater decrease in the vasodilator response of BAY 41-2272 (Emax = 20.17 +/- 4.55%; n = 6). K+-free solution also attenuated (Emax = 39.97 +/- 3.52%; n = 6) BAY 41-2272-induced relaxation. ODQ (10 microM) plus 1 microM ouabain abolished the relaxant response of BAY 41-2272 (Emax = 12.09 +/- 3.76%, n = 6 versus vehicle control dimethyl sulfoxide; Emax = 15.83 +/- 1.72%, n = 6). KT-5823 [1-oxo-9.12-epoxy-1H-diindolo[1,2,3-fg:3',2',1' kl]pyrrolo[3,4-I][1,6]benzodiazocine-10-carboxylic acid methyl ester (2 microM), a specific inhibitor of protein kinase G had no effect on 10 microM ODQ insensitive relaxation evoked by BAY 41-2272. BAY 41-2272 (10 microM) inhibited Ca2+-induced contractions in K+-depolarized preparations. BAY 41-2272 (10 microM) caused about a 14-fold increase in the intracellular cGMP over the basal level, which was completely inhibited by 10 microM ODQ. BAY 41-2272 (0.1, 1.0, and 10 microM) significantly (P < 0.05) increased ouabain-sensitive 86Rb uptake in a concentration-dependent manner. BAY 41-2272 (10 microM) also stimulated sarcolemmal Na+-K+-ATPase activity. However, 10 microM ODQ had no significant effect on either basal or BAY 41-2272-stimulated 86Rb uptake/Na+-K+-ATPase activities. In conclusion, this study provides the first evidence of sodium pump stimulation by BAY 41-2272 independent of cGMP as an additional mechanism to sGC activation in relaxation of ovine pulmonary artery. PMID- 15792997 TI - Interactions between delta and mu opioid agonists in assays of schedule controlled responding, thermal nociception, drug self-administration, and drug versus food choice in rhesus monkeys: studies with SNC80 [(+)-4-[(alphaR)-alpha ((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N diethylbenzamide] and heroin. AB - Interactions between delta and mu opioid agonists in rhesus monkeys vary as a function of the behavioral endpoint. The present study compared interactions between the delta agonist SNC80 [(+)-4-[(alphaR)-alpha-((2S,5R)-4-allyl-2,5 dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide] and the mu agonist heroin in assays of schedule-controlled responding, thermal nociception, and drug self-administration. Both SNC80 (ED50 = 0.43 mg/kg) and heroin (ED50 = 0.088 mg/kg) produced a dose-dependent and complete suppression of response rates in the assay of schedule-controlled responding. Heroin also produced thermal antinociception (ED(5 degrees C) = 0.18 mg/kg) and maintained drug self administration under both a fixed ratio schedule [dose-effect curve peak at 0.0032 mg/kg/injection (inj)] and under a food versus heroin concurrent-choice schedule (ED50 = 0.013 mg/kg/inj), whereas SNC80 did not produce thermal antinociception or maintain self-administration. Fixed ratio mixtures of SNC80 and heroin (1.6:1, 4.7:1, and 14:1 SNC80/heroin) produced additive effects in the assay of schedule-controlled responding and superadditive effects in the assay of thermal nociception. Also, SNC80 did not enhance the reinforcing effects of heroin, indicating that mixtures of SNC80 and heroin produced additive or infra additive reinforcing effects. These results provide additional evidence to suggest that delta/mu interactions depend on the experimental endpoint and further suggest that delta agonists may selectively enhance the antinociceptive effects of mu agonists while either not affecting or decreasing the sedative and reinforcing effects of mu agonists. PMID- 15792998 TI - Estimating medium- and long-term trends in malaria transmission by using serological markers of malaria exposure. AB - The implementation and evaluation of malaria control programs would be greatly facilitated by new tools for the rapid assessment of malaria transmission intensity. Because acquisition and maintenance of antimalarial antibodies depend on exposure to malaria infection, such antibodies might be used as proxy measures of transmission intensity. We have compared the prevalence of IgG antibodies with three Plasmodium falciparum asexual stage antigens in individuals of all ages living at varying altitudes encompassing a range of transmission intensities from hyper- to hypoendemic in northeastern Tanzania, with alternative measures of transmission intensity. The prevalence of antibodies to merozoite surface protein 1(19) was significantly more closely correlated with altitude than either point prevalence malaria parasitemia or single measures of hemoglobin concentration. Analysis of age-specific seroprevalence rates enabled differentiation of recent (seasonal) changes in transmission intensity from longer-term transmission trends and, using a mathematical model of the annual rate of seroconversion, estimation of the longevity of the antibody response. Thus, serological tools allow us to detect variations in malaria transmission over time. Such tools will be invaluable for monitoring trends in malaria endemicity and the effectiveness of malaria control programs. PMID- 15792999 TI - Commitment toward the natural T (iNKT) cell lineage occurs at the CD4+8+ stage of thymic ontogeny. AB - T lineage commitment occurs in a discrete, stage-specific manner during thymic ontogeny. Intrathymic precursor transfer experiments and the identification of CD4(+)8+ double-positive (DP), V alpha 14J alpha 18 natural T (iNKT) cells suggest that commitment to this lineage might occur at the DP stage. Nevertheless, this matter remains contentious because others failed to detect V alpha 14J alpha 18-positive iNKT cells that are CD4(+)8+. In resolution to this issue, we demonstrate that retinoic acid receptor-related orphan receptor gamma (ROR gamma)0/0 thymi, which accumulate immature single-positive (ISP) thymocytes that precede the DP stage, do not rearrange V alpha 14-to-J alpha 18 gene segments, suggesting that this event occurs at a post-ISP stage. Mixed radiation bone marrow chimeras revealed that RORgamma functions in an iNKT cell lineage specific manner. Further, introgression of a Bcl-x(L) transgene into ROR gamma(0/0) mice, which promotes survival and permits secondary rearrangements of distal V alpha and J alpha gene segments at the DP stage, rescues V alpha 14-to-J alpha 18 recombination. Similarly, introgression of a rearranged V alpha 14J alpha 18 transgene into ROR gamma(0/0) mice results in functional iNKT cells. Thus, our data support the "T cell receptor-instructive (mainstream precursor) model" of iNKT cell lineage specification where V alpha 14-to-J alpha 18 rearrangement, positive selection, and iNKT cell lineage commitment occur at or after the DP stage of ontogeny. PMID- 15793000 TI - Immunological role of neuronal receptor vanilloid receptor 1 expressed on dendritic cells. AB - Capsaicin (CP), the pungent component of chili pepper, acts on sensory neurons to convey the sensation of pain. The CP receptor, vanilloid receptor 1 (VR1), has been shown to be highly expressed by nociceptive neurons in dorsal root and trigeminal ganglia. We demonstrate here that the dendritic cell (DC), a key cell type of the vertebrate immune system, expresses VR1. Engagement of VR1 on immature DCs such as by treatment with CP leads to maturation of DCs as measured by up-regulation of antigen-presenting and costimulatory molecules. This effect is present in DCs of VR1+/+ but not VR1-/- mice. In VR1+/+ mice, this effect is inhibited by the VR1 antagonist capsazepine. Further, intradermal administration of CP leads to migration of DCs to the draining lymph nodes in VR1+/+ but not VR1 /- mice. These data demonstrate a powerful influence of a neuroactive ligand on a central aspect of immune function and a commonality of mechanistic pathways between neural and immune functions. PMID- 15793001 TI - Varicelloviruses avoid T cell recognition by UL49.5-mediated inactivation of the transporter associated with antigen processing. AB - Detection and elimination of virus-infected cells by cytotoxic T lymphocytes depends on recognition of virus-derived peptides presented by MHC class I molecules. A critical step in this process is the translocation of peptides from the cytoplasm into the endoplasmic reticulum by the transporter associated with antigen processing (TAP). Here, we identified the bovine herpesvirus 1-encoded UL49.5 protein as a potent inhibitor of TAP. The expression of UL49.5 results in down-regulation of MHC class I molecules at the cell surface and inhibits detection and lysis of the cells by cytotoxic T lymphocytes. UL49.5 homologs encoded by two other varicelloviruses, pseudorabies-virus and equine herpesvirus 1, also block TAP. Homologs of UL49.5 are widely present in herpesviruses, acting as interaction partners for glycoprotein M, but in several varicelloviruses UL49.5 has uniquely evolved additional functions that mediate its participation in TAP inhibition. Inactivation of TAP by UL49.5 involves two events: inhibition of peptide transport through a conformational arrest of the transporter and degradation of TAP by proteasomes. UL49.5 is degraded along with TAP via a reaction that requires the cytoplasmic tail of UL49.5. Thus, UL49.5 represents a unique immune evasion protein that inactivates TAP through a unique two-tiered process. PMID- 15793002 TI - Phosphatidylinositol phosphate kinase type I gamma regulates dynamics of large dense-core vesicle fusion. AB - Phosphatidylinositol-4,5-bisphosphate was proposed to be an important regulator of large dense-core vesicle exocytosis from neuroendocrine tissues. Here, we have examined the kinetics of secretion in chromaffin cells from mice lacking phosphatidylinositol phosphate kinase type I gamma, the major neuronal phosphatidylinositol-4-phosphate 5-kinase. Absence of this enzyme caused a reduction of the readily releasable vesicle pool and its refilling rate, with a small increase in morphologically docked vesicles, indicating a defect in vesicle priming. Furthermore, amperometry revealed a delay in fusion pore expansion. These results provide direct genetic evidence for a key role of phosphatidylinositol-4,5-bisphosphate synthesis in the regulation of large dense core vesicle fusion dynamics. PMID- 15793003 TI - A protein folding pathway with multiple folding intermediates at atomic resolution. AB - Using native-state hydrogen-exchange-directed protein engineering and multidimensional NMR, we determined the high-resolution structure (rms deviation, 1.1 angstroms) for an intermediate of the four-helix bundle protein: Rd apocytochrome b562. The intermediate has the N-terminal helix and a part of the C terminal helix unfolded. In earlier studies, we also solved the structures of two other folding intermediates for the same protein: one with the N-terminal helix alone unfolded and the other with a reorganized hydrophobic core. Together, these structures provide a description of a protein folding pathway with multiple intermediates at atomic resolution. The two general features for the intermediates are (i) native-like backbone topology and (ii) nonnative side-chain interactions. These results have implications for important issues in protein folding studies, including large-scale conformation search, -value analysis, and computer simulations. PMID- 15793004 TI - Detection of chiral sum frequency generation vibrational spectra of proteins and peptides at interfaces in situ. AB - In this work, we demonstrate the feasibility to collect off-electronic resonance chiral sum frequency generation (SFG) vibrational spectra from interfacial proteins and peptides at the solid/liquid interface in situ. It is difficult to directly detect a chiral SFG vibrational spectrum from interfacial fibrinogen molecules. By adopting an interference enhancement method, such a chiral SFG vibrational spectrum can be deduced from interference spectra between the normal achiral spectrum and the chiral spectrum. We found that the chiral SFG vibrational spectrum of interfacial fibrinogen was mainly contributed by the beta sheet structure. For a beta-sheet peptide tachyplesin I, which may be quite ordered at the solid/liquid interface, chiral SFG vibrational spectra can be collected directly. We believe that these chiral signals are mainly contributed by electric dipole contributions, which can dominate the chiroptical responses of uniaxial systems. For the first time, to our knowledge, this work indicates that the off-electronic resonance SFG technique is sensitive enough to collect chiral SFG vibrational spectra of interfacial proteins and peptides, providing more structural information to elucidate interfacial protein and peptide structures. PMID- 15793005 TI - LPS induces the interaction of a transcription factor, LPS-induced TNF-alpha factor, and STAT6(B) with effects on multiple cytokines. AB - TNF-alpha is a pivotal cytokine whose overproduction can be lethal. Previously, we identified a transcription factor, LPS-induced TNF-alpha factor (LITAF), that regulates TNF-alpha transcription. We now report the discovery and characterization of a regulatory cofactor that we call signal transducer and activator of transcription (STAT) 6(B) because of its considerable homology to STAT6 [here referred to as STAT6(A)]. The STAT6(B) gene expression was found to be activated by LPS. Furthermore, we show that cotransfection of STAT6(B) and LITAF induces an interaction between the two proteins, consequently forming a complex that subsequently translocates into the nucleus and up-regulates the transcription of cytokines. The effect of the complex on a panel of cytokines was tested. In addition, the specific role of LITAF in this complex was established with experiments, including RNA interference technology. Overall, these findings describe roles for LITAF, STAT6(B), and the LITAF-STAT6(B) complex in the regulation of inflammatory cytokines in response to LPS stimulation in mammalian cells. PMID- 15793006 TI - Three distinct kinetic groupings of the synaptotagmin family: candidate sensors for rapid and delayed exocytosis. AB - Synaptotagmins (syts) are a family of membrane proteins present on a variety of intracellular organelles. In vertebrates, 16 isoforms of syt have been identified. The most abundant isoform, syt I, appears to function as a Ca2+ sensor that triggers the rapid exocytosis of synaptic vesicles from neurons. The functions of the remaining syt isoforms are less well understood. The cytoplasmic domain of syt I binds membranes in response to Ca2+, and this interaction has been proposed to play a key role in secretion. Here, we tested the Ca(2+) triggered membrane-binding activity of the cytoplasmic domains of syts I-XII; eight isoforms tightly bound to liposomes that contained phosphatidylserine as a function of the concentration of Ca2+. We then compared the disassembly kinetics of Ca2+.syt.membrane complexes upon rapid mixing with excess Ca2+ chelator and found that syts can be classified into three distinct kinetic groups. syts I, II, and III constitute the fast group; syts V, VI, IX, and X make up the medium group; and syt VII exhibits the slowest kinetics of disassembly. Thus, isoforms of syt, which have much slower disassembly kinetics than does syt I, might function as Ca2+ sensors for asynchronous release, which occurs after Ca2+ domains have collapsed. We also compared the temperature dependence of Ca2+.syt.membrane assembly and disassembly reactions by using squid and rat syt I. These results indicate that syts have diverged to release Ca2+ and membranes with distinct kinetics. PMID- 15793007 TI - The role of binding site cluster strength in Bicoid-dependent patterning in Drosophila. AB - The maternal morphogen Bicoid (Bcd) is distributed in an embryonic gradient that is critical for patterning the anterior-posterior (AP) body plan in Drosophila. Previous work identified several target genes that respond directly to Bcd dependent activation. Positioning of these targets along the AP axis is thought to be controlled by cis-regulatory modules (CRMs) that contain clusters of Bcd binding sites of different "strengths." Here we use a combination of Bcd-site cluster analysis and evolutionary conservation to predict Bcd-dependent CRMs. We tested 14 predicted CRMs by in vivo reporter gene assays; 11 show Bcd-dependent activation, which brings the total number of known Bcd target elements to 21. Some CRMs drive expression patterns that are restricted to the most anterior part of the embryo, whereas others extend into middle and posterior regions. However, we do not detect a strong correlation between AP position of target gene expression and the strength of Bcd site clusters alone. Rather, we find that binding sites for other activators, including Hunchback and Caudal correlate with CRM expression in middle and posterior body regions. Also, many Bcd-dependent CRMs contain clusters of sites for the gap protein Kruppel, which may limit the posterior extent of activation by the Bcd gradient. We propose that the key design principle in AP patterning is the differential integration of positive and negative transcriptional information at the level of individual CRMs for each target gene. PMID- 15793008 TI - Sites of proteolytic processing and noncovalent association of the distal C terminal domain of CaV1.1 channels in skeletal muscle. AB - In skeletal muscle cells, voltage-dependent potentiation of Ca2+ channel activity requires phosphorylation by cAMP-dependent protein kinase (PKA) anchored via an A kinase anchoring protein (AKAP15), and the most rapid sites of phosphorylation are located in the C-terminal domain. Surprisingly, the site of interaction of the complex of PKA and AKAP15 with the alpha1-subunit of Ca(V)1.1 channels lies in the distal C terminus, which is cleaved from the remainder of the channel by in vivo proteolytic processing. Here we report that the distal C terminus is noncovalently associated with the remainder of the channel via an interaction with a site in the proximal C-terminal domain when expressed as a separate protein in mammalian nonmuscle cells. Deletion mapping of the C terminus of the alpha1-subunit using the yeast two-hybrid assay revealed that a distal C-terminal peptide containing amino acids 1802-1841 specifically interacts with a region in the proximal C terminus containing amino acid residues 1556-1612. Analysis of the purified alpha1-subunit of Ca(V)1.1 channels from skeletal muscle by saturation sequencing of the intracellular peptides by tandem mass spectrometry identified the site of proteolytic processing as alanine 1664. Our results support the conclusion that a noncovalently associated complex of the alpha1-subunit truncated at A1664 with the proteolytically cleaved distal C-terminal domain, AKAP15, and PKA is the primary physiological form of Ca(V)1.1 channels in skeletal muscle cells. PMID- 15793009 TI - Cortical, thalamic, and hypothalamic responses to cooling and warming the skin in awake humans: a positron-emission tomography study. AB - Thermoregulatory mechanisms are remarkably efficient, ensuring minimal temperature variation within the core of the human body under physiological conditions. Diverse afferent and efferent neural pathways contribute to the monitoring of core and skin temperature, generation of heat, and control of thermal exchange with the external environment. We have investigated the cortical, thalamic, and hypothalamic responses to cooling and warming by using positron-emission tomography activation imaging of subjects clad in a water perfused suit, which enabled rapid change of their skin-surface temperature. Human brain regions that respond to changes in skin temperature have been identified in the somatosensory cortex, insula, anterior cingulate, thalamus, and hypothalamus, with evidence that the hypothalamic response codes for the direction of temperature change. We conclude that signals from thermosensors in the skin providing crucial afferent information to the brain are integrated with signals from central thermosensors, resulting in thermoregulatory responses that maintain core temperature within a remarkably narrow range. PMID- 15793010 TI - Attention-driven discrete sampling of motion perception. AB - In movies or on TV, a wheel can seem to rotate backwards, due to the temporal subsampling inherent in the recording process (the wagon wheel illusion). Surprisingly, this effect has also been reported under continuous light, suggesting that our visual system, too, might sample motion in discrete "snapshots." Recently, these results and their interpretation have been challenged. Here, we investigate the continuous wagon wheel illusion as a form of bistable percept. We observe a strong temporal frequency dependence: the illusion is maximal at alternation rates around 10 Hz but shows no spatial frequency dependence. We introduce an objective method, based on unbalanced counterphase gratings, for measuring this phenomenon and demonstrate that the effect critically depends on attention: the continuous wagon wheel illusion was almost abolished in the absence of focused attention. A motion-energy model, coupled with attention-dependent temporal subsampling of the perceptual stream at rates between 10 and 20 Hz, can quantitatively account for the observed data. PMID- 15793011 TI - What community-based education can teach tomorrow's doctors. PMID- 15793012 TI - Community-engaged scholarship: is faculty work in communities a true academic enterprise? AB - Since Ernest Boyer's landmark 1990 report, Scholarship Reconsidered: Priorities of the Professoriate, leaders in higher education, including academic medicine, have advocated that faculty members apply their expertise in new and creative ways in partnership with communities. Such community engagement can take many forms, including community-based teaching, research, clinical care, and service. There continues to be a gap, however, between the rhetoric of this idea and the reality of how promotion and tenure actually work in health professions schools. The Commission on Community-Engaged Scholarship in the Health Professions was established in October 2003 with funding from the W.K. Kellogg Foundation to take a leadership role in creating a more supportive culture and reward system for community-engaged faculty in the nation's health professions schools. The authors prepared this article to inform the commission's deliberations and to stimulate discussion among educators in the health professions. The authors define the work that faculty engage in with communities, consider whether all work by faculty in community-based settings is actually scholarship, and propose a framework for documenting and assessing community-engaged scholarship for promotion and tenure decisions. They conclude with recommendations for change in academic health centers and health professions schools. PMID- 15793013 TI - Recognizing the value of community involvement by AHC faculty: a case study. AB - Physicians seek connections to their communities. Some health care and academic leaders believe that facilitating the creation of more such community connections is one way to reverse the trend of waning social and political legitimacy for the U.S. medical profession. For academic health centers (AHCs), such connections can maintain local and state support crucial to their long-term success. Multiple barriers exist to such involvement, especially for physicians in AHCs, where work done beyond direct patient care, administration, and research rarely contributes to the tenure and promotion process. The authors present a case study to show how one department in an AHC, beginning in the late 1990s, has been overcoming these barriers to incorporate the scholarship of community engagement into its mission and structure. The case study incorporates theoretical underpinnings to crystallize the following lessons that the department has learned so far: (1) If academic departments wish community service to be a central part of their mission, they need ways to institutionalize community engagement within organizational structures. (2) Community engagement can be scholarly. (3) If faculty members are to be recognized for their service activities, measures are necessary to determine what constitutes "excellence" and "scholarship" in community service. (4) Scholarship of community engagement goes beyond performing service activities in the community. PMID- 15793014 TI - How a community-based organization and an academic health center are creating an effective partnership for training and service. AB - Community-academic partnerships in the training of doctors offer unique learning opportunities of great importance. Such partnerships can induce a paradigm shift such that physicians view community as a teaching resource and partner rather than as a passive recipient of services or solely as a placement site. The authors describe a model of a community-academic partnership in New York City, begun in 1995, in which, for training and service, pediatric residents are integrally involved in a community-based program. Principles adapted from the Community-Campus Partnerships for Health's principles of partnership provide a framework for portraying the essential elements of developing and maintaining the partnership. The authors explain the clashes that may arise between partners and show how the principles of partnership guide partnership members in working and learning within a setting that by its nature entails conflict and inequality. This report is based on the knowledge gained from the structured reflections of both members of this partnership: the residency program at a large academic health center and the community-based social service organization. Such partnerships provide the training ground for the development of physicians who understand the social and cultural determinants of health and constructively use community agencies' input in promoting child health and well-being. Within this framework, community-based organizations are not solely service providers but become educators of physicians-in-training who, with new knowledge gained through the partnership, more effectively contribute to the overall health of the communities they serve. PMID- 15793015 TI - A historical overview of interdisciplinary family health: a community-based interprofessional health professions course. AB - The Interdisciplinary Family Health course at the University of Florida Health Science Center is a course for beginning health profession students designed to teach core values, such as community-based family health, health promotion and disease prevention, and teamwork in the context of home visits. In addition, the course provides a valuable service to volunteer families by helping them identify useful community resources, and by formulating wellness care plans for prevention of illness and stabilization of chronic illness. In this article, the authors describe the historical development of the course, which began as a grant supported pilot course for 20 medical students in 1996. After several additional grants helped fund an expansion involving other colleges, the course was given institutional support in 2001 and currently includes over 400 students and 70 faculty from four colleges working to improve the health status of over 150 local volunteer families. The theoretical constructs and objectives of the course were developed collaboratively by dedicated faculty from five Health Science Center colleges over seven years. In addition to benefiting the community and students, the course has encouraged an atmosphere of collaboration among faculty and colleges that has been a tangible benefit to the academic health center. The development and continuing support of this course demonstrates that barriers to such efforts can be overcome by dedicated faculty and administration. PMID- 15793016 TI - Teaching professionalism within a community context: perspectives from a national demonstration project. AB - Most medical schools now include some component of professionalism in their curriculum, ranging from "white coat" ceremonies to didactic and small-group, case-based discussions. Often this format does not provide a context for the course content nor does it necessarily make the curricular themes relevant to population groups and communities most vulnerable to the inequities and injustices present in health care. The authors describe a community-based professionalism curriculum for preclinical and clinical year medical students and report evaluation data from three years (2001-2003) of this national demonstration project. The curriculum emphasized four themes: service, community, advocacy, and ethical behavior and was based on a service-learning pedagogy applied within community-based organizations. As part of the program evaluation, 95 students from 33 medical schools between the years 2001 and 2003 (response rate: 84.8%) completed an anonymous questionnaire. When asked what did they learn about professionalism that they did not learn (or expect to learn) in their medical school curriculum, the most common themes were (1) factors and influences affecting professional behavior, with many specifically citing pharmaceutical companies and insurance carriers (46.3%); (2) the role and importance of physician advocacy on behalf of their patients (37.9%); and (3) issues specific to the needs of vulnerable and disadvantaged populations (20.0%). This project demonstrates that community-based experiences can provide unique and relevant learning in a professionalism curriculum that can complement existing medical school-based efforts. PMID- 15793017 TI - Informal mentoring between faculty and medical students. AB - Mentoring skills are valuable assets for academic medicine faculty, who help shape the professionalism of the next generation of physicians. Mentors are role models who also act as guides for students' personal and professional development over time. Mentors can be instrumental in conveying explicit academic knowledge required to master curriculum content. Importantly, they can enhance implicit knowledge about the "hidden curriculum" of professionalism, ethics, values and the art of medicine not learned from texts. In many cases, mentors also provide emotional support and encouragement. The relationship benefits mentors as well, through greater productivity, career satisfaction, and personal gratification. Maximizing the satisfaction and productivity of such relationships entails self awareness, focus, mutual respect, and explicit communication about the relationship. In this article, the authors describe the development of optimal mentoring relationships, emphasizing the importance of experience and flexibility in working with beginning to advanced students of different learning styles, genders, and races. Concrete advice for mentor "do's and don'ts"is offered, with case examples illustrating key concepts. PMID- 15793018 TI - The importance of anatomy in health professions education and the shortage of qualified educators. AB - The current shortage of faculty qualified to teach anatomy in U.S. medical schools is reversible. Sufficient numbers of individuals are in the pipeline to provide a future cadre of well-trained faculty members educating students in gross anatomy. The challenge is to realign departmental, institutional, and federal training grant priorities and resources, creating incentives for graduate students, postdoctoral fellows, and faculty members to stay the course and become the teachers needed to educate the next generation of health professionals. These strategies include (but are not limited to) team-teaching gross anatomy, thereby distributing the time commitments of a laboratory-based course more widely within a department; funds made available from the administration of medical schools to allow postdoctoral fellows to participate in teaching and providing compensation for the research activities; using "mission-based budgeting" to specifically compensate for faculty teaching time; and, finally, re-instituting federally funded training grants that solved this same teaching crisis in the not-too distant past. PMID- 15793019 TI - A course on the transition to adult care of patients with childhood-onset chronic illnesses. AB - Children with special health care needs born today have a 90% chance of surviving into adulthood, making their transition to adult systems of care an issue that will affect almost all physicians. However, many adult generalists and specialists are not familiar with the management of chronic diseases that begin in childhood. While the public health system has made transition to appropriate adult care a priority, and many specialty organizations have endorsed this concept, there are no published studies addressing how the concept of transition can be taught to medical students or residents. The authors describe a one-week course for medical students, begun in 2001 at their institution, that addresses the transition for youth with special health care needs, emphasizing patient and family-centered care, cultural competence, and decision making in end-of-life issues. Cystic fibrosis, a common genetic disease with increasing life expectancy, is used as the model for the course. Involvement of interdisciplinary faculty, interviews with youth with special health care needs and family caregivers, readings from academic and nonacademic literature, and group discussions are presented as teaching methods. Key insights based on experience with the course are the need to include the voices of patients and families, the use of faculty from various professions and specialties to model interdisciplinary care, and the insight that problems specific to transition offer into contemporary health care financing. Future studies should measure the impact of such courses on students' knowledge of transition issues, and determine essential information required for physicians in practice. PMID- 15793020 TI - Corridors. PMID- 15793021 TI - The emergence of academies of educational excellence: a survey of U.S. medical schools. AB - PURPOSE: To identify existing organizations that recognize faculty members' excellence as educators (Academies) in the United States, and describe the organizations' characteristics. METHOD: A 31-item questionnaire inquiring about Academies or equivalent programs was sent to deans of medical education at all 125 U.S. medical schools in February of 2003. Variables examined were general Academy characteristics such as membership selection criteria, goals, benefits of membership, and budget, as well as, estimates of prestige of membership and influence on recruiting new and current faculty to educational activities. RESULTS: Twenty of 97 (21%) respondents reported an implemented Academy or equivalent program (eight begun prior to 2000 and 11 subsequently). Most Academies (75%) did not "cap" membership size, and most (65%) offered lifetime membership. Budgets ranged from 0 dollars to more than 100,000 dollars per year. Full-time faculty status (100%) and involvement in direct undergraduate teaching (95%) affected eligibility the most. Nominations for membership most often came from department or section chairs (89%) and from peers (74%), and learners were involved in the final selection process at 18 of the Academies. Benefits of membership included networking/collaboration, school-wide recognition, and mentoring for educational skills development. The benefit of protected time was offered at only three institutions and was associated with having a larger budget. Respondents believe Academies positively influence faculty participation in educational activities. CONCLUSIONS: Academies are formal organizations recognizing faculty contributions to medical education, and they are increasing in number. They offer important benefits to faculty members and the educational mission of an academic medical center. PMID- 15793022 TI - Measuring professionalism: a review of studies with instruments reported in the literature between 1982 and 2002. AB - PURPOSE: To describe the measurement properties of instruments reported in the literature that faculty might use to measure professionalism in medical students and residents. METHOD: The authors reviewed studies published between 1982 and 2002 that had been located using Medline and four other databases. A national panel of 12 experts in measurement and research in medical education extracted data from research reports using a structured critique form. RESULTS: A total of 134 empirical studies related to the concept of professionalism were identified. The content of 114 involved specific elements of professionalism, such as ethics, humanism, and multiculturalism, or associated phenomena in the educational environment such as abuse and cheating. Few studies addressed professionalism as a comprehensive construct (11 studies) or as a distinct facet of clinical competence (nine studies). The purpose of 109 studies was research or program evaluation, rather than summative or formative assessment. Sixty five used self administered instruments with no independent observation of the participants' professional behavior. Evidence of reliability was reported in 62 studies. Although content validity was reported in 86 studies, only 34 provided strong evidence. Evidence of concurrent or predictive validity was provided in 43 and 16 studies, respectively. CONCLUSIONS: There are few well-documented studies of instruments that can be used to measure professionalism in formative or summative evaluation. When evaluating the tools described in published research it is essential for faculty to look critically for evidence related to the three fundamental measurement properties of content validity, reliability, and practicality. PMID- 15793023 TI - "Once when i was on call...," theory versus reality in training for professionalism. AB - PURPOSE: To identify the degree to which interns' reported experiences with professional and unprofessional behavior converge and/or diverge with ideal professional behavior proposed by the physician community. METHOD: Interns at Wayne State University's residency programs in internal medicine, family medicine, and transitional medicine responded to essay questions about their experience with professional and unprofessional behavior as part of a curriculum on professionalism. Responses were coded for whether they reflected each of the principles and responsibilities outlined in a major publication on physician professionalism. Content analysis included the frequencies with which the interns' essays reflected each principle or responsibility. Additionally, a thematic analysis revealed themes of professional behavior that emerged from the essays. RESULTS: Interns' experiences with professional and unprofessional behavior most frequently converged with ideal behavior proposed by the physician community in categories involving interpersonal interactions with patients. Interns infrequently reported experiences involving behavior related to systems or sociopolitical issues. CONCLUSIONS: Interns' essays reflect their concern with interpersonal interactions with patients, but they are either less exposed to or less interested in describing behavior regarding systems or sociopolitical issues. This may be due to their stage of training or to the emphasis placed on interpersonal rather than systems or sociopolitical issues during training. The authors recommend future proposals of ideal professional behavior be revised periodically to reflect current experiences of practicing physicians, trainees, other health care providers and patients. Greater educational emphasis should be placed on the systems and sociopolitical environment in which trainees practice. PMID- 15793024 TI - Factors affecting resident performance: development of a theoretical model and a focused literature review. AB - PURPOSE: The clinical performances of physicians have come under scrutiny as greater public attention is paid to the quality of health care. However, determinants of physician performance have not been well elucidated. The authors sought to develop a theoretical model of physician performance, and explored the literature about factors affecting resident performance. METHOD: Using expert consensus panel, in 2002-03 the authors developed a hypothesis-generating model of resident performance. The developed model had three input factors (individual resident factors, health care infrastructure, and medical education infrastructure), intermediate process measures (knowledge, skills, attitudes, habits), and final health outcomes (affecting patient, community and population). The authors used factors from the model to focus a PubMed search (1967-2002) for all original articles related to the factors of individual resident performance. RESULTS: The authors found 52 original studies that examined factors of an individual resident's performance. They describe each study's measurement instrument, study design, major findings, and limitations. Studies were categorized into five domains: learning styles/personality, social/financial factors, practice preferences, personal health, and response to job environment. Few studies examined intermediate or final performance outcomes. Most were single institution, cross-sectional, and survey-based studies. CONCLUSIONS: Attempting to understand resident performance without understanding factors that influence performance is analogous to examining patient adherence to medication regimens without understanding the individual patient and his or her environment. Based on a systematic review of the literature, the authors found few discrete associations between the factors of individual resident and the resident's actual job performance. Additionally, they identify and discuss major gaps in the educational literature. PMID- 15793025 TI - Use of an electronic medical record to profile the continuity clinic experiences of primary care residents. AB - PURPOSE: To assess the variation in outpatient educational experiences among residents in a primary care internal medicine residency program. METHOD: The authors conducted an observational study within a primary care residency program in Boston, Massachusetts, involving eight primary care residents in 2001-02. A data management system was created that uses information on clinical experiences collected from an electronic medical record (EMR). The EMR records clinical information from patient encounters in resident continuity clinics, including patient demographics and diagnostic codes entered by the residents. Primary and secondary diagnosis from the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) were assigned to clinical categories (e.g., cardiology, orthopedics) and these assignments were programmed into a spreadsheet that could take the diagnostic data directly from the EMR downloads and produce reports using only the primary diagnosis, using all diagnoses, or using the assigned clinical categories. RESULTS: The EMR download showed 2,468 patient encounters for the eight residents in the primary care program. The most common clinical encounters were for health maintenance (17%), cardiology (14%), and women's health (10%). In contrast, rheumatology (0.5%) and hematology/oncology (2%) encounters were less frequent. There were substantial variations among residents in terms of distribution of encounter diagnoses, and the age and gender of patients seen in continuity clinic. CONCLUSIONS: Abstracting data from an EMR represents a feasible method for assessing programmatic and individual learner experiences in the outpatient setting. Such information may help target curricular adjustments to ensure an appropriate diversity and depth of clinical training. PMID- 15793026 TI - The psychometric properties of five scoring methods applied to the script concordance test. AB - PURPOSE: The Script Concordance Test (SCT) is designed to measure cognitive ability related to successful clinical decision making. An SCT's usefulness for medical education depends on establishing its construct validity. The SCT's present construct relates examinee's scores to experts' response patterns, which does not require a single-best-answer format. Because medical education assessments do require a single best answer, the authors compared the psychometric properties of two aggregate scoring methods with three single-best answer scoring methods for an SCT. METHOD: A nephrology SCT was developed and administered to 85 examinees. Examinees' scores derived from a key developed using eight experts and a traditional aggregate scoring method on a five-point Likert-based scale were compared with four alternate scoring methods (one method eliminated the multipoint Likert-type scale and three eliminated the Likert-type scale and employed single-best-answer scoring). RESULTS: Two of the four alternate scoring methods performed as well as the traditional Likert-type aggregate scoring method. Scores from all five methods were highly intercorrelated. In addition, each method produced scores similarly correlated with level of experience, and none exhibited an intermediate effect. CONCLUSIONS: Single-best-answer scoring with three answer choices produced results similar to aggregate scoring on a Likert-type scale. Because SCT items appear to assess an examinee's understanding of the interrelatedness of medical knowledge, single best-answer scoring on an SCT may be valid as an educational assessment. More research is needed to assess differential validity compared with multiple-choice question exams and the predictive validity related to clinical performance. PMID- 15793028 TI - An introduction to foreign health care. PMID- 15793027 TI - Does students' exposure to gender discrimination and sexual harassment in medical school affect specialty choice and residency program selection? AB - PURPOSE: To examine the role of gender discrimination and sexual harassment in medical students' choice of specialty and residency program. METHOD: Anonymous, self-administered questionnaires were distributed in 1997 to fourth-year students enrolled in 14 public and private U.S. medical schools. In addition to reporting the frequency of gender discrimination and sexual harassment encountered during preclinical coursework, core clerkships, elective clerkships, and residency selection, students assessed the impact of these exposures (none, a little, some, quite a bit, the deciding factor) on their specialty choices and rankings of residency programs. RESULTS: A total of 1,314 (69%) useable questionnaires were returned. Large percentages of men (83.2%) and women (92.8%) experienced, observed, or heard about at least one incident of gender discrimination and sexual harassment during medical school, although more women reported such behavior across all training contexts. Compared with men, significantly (p /= 3.1 mm to group A (n = 27) and nitrite > > chloride. These currents showed little sensitivity to pH or the nature of the counter cation in the oocyte bath solution. One-half maximal currents were induced by nitrate concentrations between 1 to 3 mm. No apparent transport of organic anions was observed. Voltage clamp records of an ortholog of GmN70 from Lotus japonicus (LjN70; K. Szczyglowski, P. Kapranov, D. Hamburger, F.J. de Bruijn [1998] Plant Mol Biol 37: 651-661) also showed anion currents with a similar selectivity profile. Overall, these findings suggest that GmN70 and LjN70 are inorganic anion transporters of the symbiosome membrane with enhanced preference for nitrate. These transport activities may aid in regulation of ion and membrane potential homeostasis, possibly in response to external nitrate concentrations that are known to regulate the symbiosis. PMID- 15793075 TI - Effect of fluid ingestion on neuromuscular function during prolonged cycling exercise. AB - OBJECTIVES: To investigate the effects of fluid ingestion on neuromuscular function during prolonged cycling exercise. METHODS: Eight well trained subjects exercised for 180 minutes in a moderate environment at a workload requiring approximately 60% maximal oxygen uptake. Two conditions, fluid (F) and no fluid (NF) ingestion, were investigated. RESULTS: During maximal voluntary isometric contraction (MVC), prolonged cycling exercise reduced (p<0.05) the maximal force generating capacity of quadriceps muscles (after three hours of cycling) and root mean square (RMS) values (after two hours of cycling) with no difference between the two conditions despite greater body weight loss (p<0.05) in NF. The mean power frequency (MPF) for vastus lateralis muscle was reduced (p<0.05) and the rate of force development (RFD) was increased (p<0.05) only during NF. During cycling exercise, integrated electromyographic activity and perceived exertion were increased in both conditions (p<0.05) with no significant effect of fluid ingestion. CONCLUSIONS: The results suggest that fluid ingestion did not prevent the previously reported decrease in maximal force with exercise duration, but seems to have a positive effect on some indicators of neuromuscular fatigue such as mean power frequency and rate of force development during maximal voluntary contraction. Further investigations are needed to assess the effect of change in hydration on neural mechanisms linked to the development of muscular fatigue during prolonged exercise. PMID- 15793076 TI - Drug use in English professional football. AB - OBJECTIVES: To examine several issues related to drug use in English professional football. More particularly the project sought to gather data on: players' use of permitted supplements (mineral and vitamin pills and creatine); whether they sought advice, and if so from whom, about their use of supplements; their experience of and attitudes towards drug testing; their views on the extent of the use of banned performance enhancing and recreational drugs in football; and their personal knowledge of players who used such drugs. METHODS: With the cooperation of the Professional Footballers Association (PFA), reply paid postal questionnaires were delivered to the home addresses of all 2863 members of the PFA. A total of 706 questionnaires were returned, a response rate of just under 25%. RESULTS: Many players use supplements, although almost one in five players does so without seeking qualified professional advice from anyone within the club. Blood tests are rarely used to monitor the health of players. One third of players had not been tested for drugs within the preceding two years, and 60% felt that they were unlikely to be tested in the next year. The use of performance enhancing drugs appears to be rare, although recreational drugs are commonly used by professional footballers: 6% of respondents indicated that they personally knew players who used performance enhancing drugs, and 45% of players knew players who used recreational drugs. CONCLUSIONS: There is a need to ensure that footballers are given appropriate advice about the use of supplements in order to minimise the risk of using supplements that may be contaminated with banned substances. Footballers are tested for drugs less often than many other elite athletes. This needs to be addressed. The relatively high level of recreational drug use is not reflected in the number of positive tests. This suggests that many players who use recreational drugs avoid detection. It also raises doubts about the ability of the drug testing programme to detect the use of performance enhancing drugs. PMID- 15793077 TI - The repeatability and criterion related validity of the 20 m multistage fitness test as a predictor of maximal oxygen uptake in active young men. AB - OBJECTIVE: To investigate the repeatability and criterion related validity of the 20 m multistage fitness test (MFT) for predicting maximal oxygen uptake (Vo(2max)) in active young men. METHODS: Data were gathered from two phases using 30 subjects (x+/-s; age = 21.8+/-3.6 years, mass = 76.9+/-10.7 kg, stature = 1.76+/-0.05 m). MFT repeatability was investigated in phase 1 where 21 subjects performed the test twice. The MFT criterion validity to predict Vo(2max) was investigated in phase 2 where 30 subjects performed a continuous incremental laboratory test to volitional exhaustion to determine Vo(2max) and the MFT. RESULTS: Phase 1 showed non-significant bias between the two applications of the MFT (x(diff)+/-s(diff) = -0.4+/-1.4 ml kg(-1) min(-1); t = -1.37, p = 0.190) with 95% limits of agreement (LoA) +/-2.7 ml kg(-1) min(-1) and heteroscedasticity 0.223 (p = 0.330). Log transformation of these data reduced heteroscedasticity to 0.056 (p = 0.808) with bias -0.007+/-0.025 (t = -1.35, p = 0.190) and LoA+/ 0.049. Antilogs gave a mean bias on the ratio scale of 0.993 and random error (ratio limits) x/divided by 1.050. Phase 2 showed that the MFT significantly underpredicted Vo(2max) (x(diff)+/-s(diff) = 1.8+/-3.2 ml kg(-1) min(-1); t = 3.10, p = 0.004). LoA were +/-6.3 ml kg(-1) min(-1) and heteroscedasticity 0.084 (p = 0.658). Log transformation reduced heteroscedasticity to -0.045 (p = 0.814) with LoA+/-0.110. The significant systematic bias was not eliminated (x(diff)+/ s(diff) = 0.033+/-0.056; t = 3.20, p = 0.003). Antilogs gave a mean bias of 1.034 with random error x/divided by 1.116. CONCLUSIONS: These findings lend support to previous investigations of the MFT by identifying that in the population assessed it provides results that are repeatable but it routinely underestimates Vo(2max) when compared to laboratory determinations. Unlike previous findings, however, these results show that when applying an arguably more appropriate analysis method, the MFT does not provide valid predictions of Vo(2max). PMID- 15793078 TI - Motivation and satisfaction among polyclinic volunteers at the 2002 Winter Olympic and Paralympic Games. AB - BACKGROUND: The Olympic and Paralympic Games rely heavily on volunteers to provide many essential services, including medical care of athletes. OBJECTIVE: This preliminary investigation sought to characterise the motivational influences and factors responsible for the satisfaction of Olympic and Paralympic healthcare volunteers. METHODS: The 2002 Winter Games polyclinic healthcare volunteers were asked to complete a questionnaire designed to elicit information about their motives for volunteering and the factors that contributed to their satisfaction with their volunteer experience. RESULTS: There was no significant difference in the motivation or satisfaction summary scores based on event worked. There was a strong positive correlation between motivation and satisfaction. Physician respondents had a lower mean motivation score than did non-physician volunteers. CONCLUSIONS: There were no significant motivational differences between Olympic and Paralympic volunteers, but there were several differences noted between physician and non-physician volunteers. The 2002 polyclinic volunteers appear to have been motivated by a complex process best described as "enlightened self interest," and all were generally well satisfied with their experience. These results may assist organisers of future Games in selecting appropriately motivated volunteer personnel and creating rewarding work environments for them. PMID- 15793080 TI - Methods for injury surveillance in international cricket. AB - BACKGROUND: The varying methods of cricket injury surveillance have made direct comparison of published studies in this field impossible. METHODS: A consensus regarding definitions and methods to calculate injury rates in cricket was sought between researchers in this field. This was arrived at through a variety of face to face meetings, email communication, and draft reviews between researchers from six of the major cricket playing nations. RESULTS: It is recommended that a cricket injury is defined as any injury or other medical condition that either (a) prevents a player from being fully available for selection for a major match or (b) during a major match, causes a player to be unable to bat, bowl, or keep wicket when required by either the rules or the team's captain. Recommended definitions for injury incidence (for matches, training sessions, and seasons) and injury prevalence are also provided. It is proposed that match injury incidence is calculated using a denominator based on a standard time estimated for player exposure in matches, for the purposes of simplicity. This will allow all injury surveillance systems, including those with limited resources, to make calculations according to a standard definition. CONCLUSION: The consensus statement presented provides a standard which, if followed, allows meaningful comparison of injury surveillance data from different countries and time periods, which will assist in the possible identification of risk factors for injury in cricket. PMID- 15793079 TI - Neck injuries presenting to emergency departments in the United States from 1990 to 1999 for ice hockey, soccer, and American football. AB - OBJECTIVE: To examine the number and rate of neck injuries in the community as a whole for ice hockey, soccer, and American football by analysing data from patients presenting to emergency departments in the United States from 1990 to 1999. METHODS: Data compiled for the US Consumer Product Safety Commission were used to generate estimates for the total number of neck injuries and the more specific diagnoses of neck fractures, dislocations, contusions, sprains, strains, and lacerations occurring nationally from 1990 to 1999. These data were combined with yearly participation figures to generate rates of injury presenting to emergency departments for each sport. RESULTS: There were an estimated 5038 neck injuries from ice hockey, 19,341 from soccer, and 114 706 from American football. These could be broken down as follows: 4964 contusions, sprains, or strains from ice hockey, 17,927 from soccer, and 104 483 from football; 105 neck fractures or dislocations from ice hockey, 214 from soccer, and 1588 from football; 199 neck lacerations for ice hockey, 0 for soccer, and 621 for football. The rates for total neck injuries and combined neck contusions, sprains, or strains were higher for football than for ice hockey or soccer in all years for which data were available. CONCLUSION: The rate of neck injury in the United States was higher in football than in ice hockey or soccer in the time period studied. PMID- 15793081 TI - Yada Yada Yada or carpe somnum. PMID- 15793082 TI - Exercises for improving bone strength. PMID- 15793084 TI - A review of the stroke volume response to upright exercise in healthy subjects. AB - Traditionally, it has been accepted that, during incremental exercise, stroke volume plateaus at 40% of Vo(2)max. However, recent research has documented that stroke volume progressively increases to Vo(2)max in both trained and untrained subjects. The stroke volume response to incremental exercise to Vo(2)max may be influenced by training status, age, and sex. For endurance trained subjects, the proposed mechanisms for the progressive increase in stroke volume to Vo(2)max are enhanced diastolic filling, enhanced contractility, larger blood volume, and decreased cardiac afterload. For untrained subjects, it has been proposed that continued increases in stroke volume may result from a naturally occurring high blood volume. However, additional research is needed to evaluate the importance of blood volume, or other mechanisms, that influence the stroke volume response to exercise in untrained subjects. PMID- 15793085 TI - Summary and agreement statement of the 2nd International Conference on Concussion in Sport, Prague 2004. AB - In November 2001, the 1st International Symposium on Concussion in Sport was held in Vienna, Austria to provide recommendations for the improvement of safety and health of athletes who suffer concussive injuries in ice hockey, football (soccer), and other sports. The 2nd International Symposium on Concussion in Sport was organised by the same group and held in Prague, Czech Republic in November 2004. It resulted in a revision and update of the Vienna consensus recommendations, which are presented here. PMID- 15793087 TI - Sweat rate and fluid turnover in American football players compared with runners in a hot and humid environment. AB - OBJECTIVES: To determine sweat rate (SwR) and fluid requirements for American footballers practicing in a hot, humid environment compared with cross country runners in the same conditions. METHODS: Fifteen subjects, 10 footballers and five runners, participated. On the 4th and 8th day of preseason two a day practices, SwR during exercise was determined in both morning and afternoon practices/runs from the change in body mass adjusted for fluids consumed and urine produced. Unpaired t tests were used to determine differences between groups. RESULTS: Overall SwR measured in litres/h was higher in the footballers than the cross country runners (2.14 (0.53) v 1.77 (0.4); p<0.01). Total sweat loss in both morning (4.83 (1.2) v 1.56 (0.39) litres) and afternoon (4.8 (1.2) v 1.97 (0.28) litres) practices/runs, and daily sweat losses (9.4 (2.2) v 3.53 (0.54) litres) were higher in the footballers (p<0.0001). The footballers consumed larger volumes of fluid during both morning and afternoon practices/runs (23.9 (8.9) v 5.5 (3.1) ml/min and 23.5 (7.3) v 13.6 (5.6) ml/min; p<0.01). For complete hydration, the necessary daily fluid consumption calculated as 130% of daily sweat loss in the footballers was 12.2 (2.9) litres compared with 4.6 (0.7) litres in the runners (p<0.0001). Calculated 24 hour fluid requirements in the footballers ranged from 8.8 to 19 litres. CONCLUSIONS: The American footballers had a high SwR with large total daily sweat losses. Consuming large volumes of hypotonic fluid may promote sodium dilution. Recommendations for fluid and electrolyte replacement must be carefully considered and monitored in footballers to promote safe hydration and avoid hyponatraemia. PMID- 15793089 TI - Injuries in women's professional soccer. AB - OBJECTIVE: The injury data from the first two seasons of the Women's United Soccer Association (WUSA) were analysed to determine the injury incidence, anatomic location of injuries, and relation of player position. METHODS: Injury data on 202 players from eight teams during the first two seasons of the WUSA were prospectively collected and analysed. RESULTS: A total of 173 injuries occurred in 110 players with an overall injury incidence rate of 1.93 injuries per 1000 player hours. The incidence of injury during practice and games was 1.17 and 12.63 per 1000 player hours, respectively. Of the injuries 82% were acute and 16% were chronic. Most of the injuries (60%) were located in the lower extremities. Strains (30.7%), sprains (19.1%), contusions (16.2%), and fractures (11.6%) were the most common diagnoses and the knee (31.8%) and head (10.9%) were the most common sites of injury. Anterior cruciate ligament (ACL) injuries accounted for 4.6% of all injuries and the incidence of ACL tears was 0.09 per 1000 player hours (practice 0.04, game 0.90). Midfielders suffered the most injuries (p<0.007). CONCLUSION: We conclude that the injury incidence in the WUSA is lower than the 6.2 injuries per 1000 player hours found in the corresponding male professional league (Major League Soccer); however, knee injuries predominate even in these elite female athletes. PMID- 15793092 TI - A controlled study on batted ball speed and available pitcher reaction time in slowpitch softball. AB - OBJECTIVES: To investigate safety risks in slowpitch softball by conducting laboratory and experimental studies on the performance of high tech softball bats with polyurethane softballs. To compare the results with the recommended safety standards. METHODS: ASTM standard compression testing of seven softball models was conducted. Using these seven softball models, bat/ball impact testing was performed using seven adult male softball players and six high tech softball bat models to determine mean batted ball speeds. Over 500 bat/ball impact measurements were recorded and analysed. Available pitcher reaction time was calculated from the mean batted ball speed measurements. RESULTS: According to the United States Specialty Sports Association and the Amateur Softball Association, the maximum initial batted ball speed should be 137.2 km/h, which corresponds to a minimum pitcher reaction time of 0.420 second. These experiments produced mean batted ball speeds of 134.0-159.7 km/h, which correspond to available pitcher reaction times of 0.409-0.361 second. CONCLUSION: The use of high tech softball bats with polyurethane softballs can result in batted ball speeds that exceed the recommended safety limits, which correspond to decreased available pitcher reaction times. PMID- 15793091 TI - Men, muscles, and body image: comparisons of competitive bodybuilders, weight trainers, and athletically active controls. AB - OBJECTIVES: To investigate body image and psychosocial adjustment among competitive bodybuilders, non-competitive weight trainers, and athletically active men. METHODS: Participants were 40 men in each of the three groups who were assessed on body composition and multiple facets of body image evaluation, investment and anxiety, eating attitudes, and social self esteem. RESULTS: Relative to the other two groups, competitive bodybuilders had greater body mass due to fat-free body mass. Although groups did not differ in their situational body image discomfort, competitive bodybuilders and weight trainers had a more positive global appearance evaluation and were more psychologically invested in their physical appearance. Compared with active controls, men in both weightlifting groups were more satisfied with their upper torso and muscle tone. Competitive bodybuilders reported more mid torso satisfaction than the other two groups. Competitive bodybuilders also wished to be significantly heavier than controls did and reported higher social self esteem but greater eating disturbance. CONCLUSIONS: The findings suggest that competitive bodybuilders as a group are not more "muscle dysmorphic" than either non-competitive weight trainers or physically active men who do not train with weights. PMID- 15793093 TI - Head injuries in youth soccer players presenting to the emergency department. AB - BACKGROUND: There has been recent concern about neuropsychological injuries experienced by soccer players, particularly related to the purposeful heading of the ball. There are few population based analyses examining whether this is a legitimate concern. OBJECTIVES: To explore, using an existing injury surveillance system, one of many parts of this issue: acute injuries requiring emergency medical care experienced by youth soccer players. METHODS: Descriptive epidemiological analysis of emergency department injury surveillance data (1996 2001) for youths aged 10-24 years from the Kingston sites of the Canadian Hospital Injury Reporting and Prevention Program. RESULTS: A total of 1714 cases of soccer injury were identified (mean 286 a year); 235 (13.7%) involved diagnoses of injuries to the head. Leading mechanical factors resulting in head injury were contact with other players or persons (153/235; 65.1%) and balls (62/235; 26.4%). Heading was reported in 4/62 (6%) of the ball contact injuries, and attempted heading was reported in 15/153 (9.8%) of the cases involving person to person contact. Unspecified head to head contact between players was reported in 39 cases. CONCLUSIONS: Minor head injuries that result in emergency medical treatment do not happen often in youth soccer, and very few can be attributed to the purposeful heading of the ball. Player contact injuries appear to be a more important injury control concern. This study informs one of many aspects of the soccer heading injury debate. PMID- 15793094 TI - Effects of montelukast on airway narrowing from eucapnic voluntary hyperventilation and cold air exercise. AB - BACKGROUND: Exercise induced bronchoconstriction (EIB) is common in elite athletes. Eucapnic voluntary hyperventilation (EVH) is a laboratory test recommended for the identification of EIB in athletes, secondary to a field exercise challenge. Montelukast attenuates EIB, but its protective effect against airway narrowing from EVH has not been investigated. OBJECTIVE: To examine the effectiveness of montelukast after exercise and after EVH. METHODS: A randomised, placebo controlled, double blind, crossover study was performed with 11 physically active EIB positive subjects (eight men, three women; mean (SD) age 22.8 (6.8) years). Six hours before each of the following challenges 10 mg montelukast or placebo was ingested: (a) a six minute, cold air (-3 degrees C) maximal effort work accumulation cycle ergometer exercise; (b) EVH, breathing 5% CO(2) compressed air at 85% maximal voluntary ventilation for six minutes. Spirometry was performed before and 5, 10, and 15 minutes after the challenge. At least 48 hours was observed between challenges. RESULTS: No differences in forced expiratory volume in one second (FEV(1)) were found after the two challenges. Exercise and EVH resulted in falls in FEV(1) of 22.4 (18.0) and 25.6 (16.8) respectively. Falls in FEV(1) after montelukast were less than after placebo (10.6 (10.6) and 14.3 (11.3) after exercise and EVH respectively; p<0.05). Montelukast provided protection against bronchoconstriction (59% and 53%; p<0.05) for eight exercising subjects and 10 EVH subjects; no protection was afforded for three exercising and one EVH challenged subject. CONCLUSIONS: Both exercise and EVH were potent stimuli of airway narrowing. A single dose of montelukast provided reasonable protection in attenuating bronchoconstriction from either exercise or EVH. The similar protection by montelukast suggests that EVH is a suitable laboratory surrogate for EIB evaluation. PMID- 15793095 TI - An assessment of injuries in college cheerleading: distribution, frequency, and associated factors. AB - BACKGROUND: Over 50% of all catastrophic injuries in women's sport occur in cheerleading, but unlike other sports, no central tracking system exists. OBJECTIVE: To obtain, describe, and compare cheerleading injury data and associated factors. METHODS: Cheerleaders from randomly chosen division IA universities completed surveys designed to acquire basic information and data on injury frequency, type, and location, practice frequency and duration, and related factors. RESULTS: Participants (n = 440) were aged 18-23 (mean (SD) 20.2 (1.8)) with 6.6 (2.2) years of experience. Most respondents (78%) reported having suffered one or more career injury. Of those injured, 39.7% reported an injury within the preceding year. Respondents sustained 1.0 (0.91) injuries during the preceding year with 1.8 (2.2) days lost. Ankles (44.9%) and wrist/hand (19.3%) were the most commonly injured. Practice frequency and duration were 205 (61.5) days a year (range 80-300) and 2.8 (0.7) hours (range 1.5-4) respectively. Training included stretching (99.7%), endurance activities (87.1%), and weight training (92.9%). CONCLUSION: Guidelines and policy governing cheerleading should be developed according to mandatory injury reporting similar to that currently used in other sports. PMID- 15793096 TI - Modafinil in sports: ethical considerations. AB - Performance enhancing agents are prohibited in athletic competition so that only athletic skills can determine outcomes. Modafinil is a novel non-addicting psychostimulant approved for treatment of narcolepsy. Does its use, especially for medical indications, violate the Olympic Movement Anti-Doping Code? This is discussed with reference to a current high profile case. PMID- 15793098 TI - Accumulation and oriented transport of ampicillin in Caco-2 cells from its pivaloyloxymethylester prodrug, pivampicillin. AB - Pivampicillin (PIVA), an acyloxymethylester of ampicillin, is thought to enhance the oral bioavailability of ampicillin because of its greater lipophilicity compared to that of ampicillin. The fate of PIVA in intestinal cells and the exact location of its conversion into ampicillin have, however, never been unambiguously established. Polarized Caco-2 cells have been used to examine the handling of PIVA and the release of ampicillin from PIVA by the intestinal epithelium. Experiments were limited to 3 h. Cells incubated with PIVA (apical pole) showed a fast accumulation of ampicillin and transport toward the basolateral medium, whereas PIVA itself was only poorly accumulated and transported. Cells incubated with free ampicillin accumulated and transported only minimal amounts of this drug. Release of ampicillin from cells incubated with PIVA was unaffected by PEPT1 and OCTN2 inhibitors but was sharply decreased after ATP depletion or addition of bis(4-nitrophenyl)-phosphate (BNPP; an esterase inhibitor). PIVA incubated with Caco-2 lysates released free ampicillin, and this release was inhibited by BNPP. Efflux studies showed that the ampicillin that accumulated in cells after incubation with PIVA was preferentially transported out of the cells through the basolateral pole. This efflux was decreased by multidrug resistance-associated protein (MRP) inhibitors (probenecid, MK-571) and by ATP depletion. A phthalimidomethylester of ampicillin that resists cellular esterases failed to cause any significant release (cell lysate) or transport (polarized Caco-2 cells) of ampicillin. These results show that when PIVA is given to Caco-2 cells from their apical pole, ampicillin is released intracellularly and that ampicillin is thereafter preferentially effluxed into the basolateral medium through an MRP-like transporter. PMID- 15793099 TI - Evidence for multiple-antibiotic resistance in Campylobacter jejuni not mediated by CmeB or CmeF. AB - An efflux system, CmeABC, in Campylobacter jejuni was previously described, and a second efflux system, CmeDEF, has now been identified. The substrates of CmeDEF include ampicillin, ethidium bromide, acridine, sodium dodecyl sulfate (SDS), deoxycholate, triclosan, and cetrimide, but not ciprofloxacin or erythromycin. C. jejuni NCTC11168 and two efflux pump knockout strains, cmeB::Kan(r) and cmeF::Kan(r), were exposed to 0.5 to 1 microg of ciprofloxacin/ml in agar plates. All mutants arising from NCTC11168 were resistant to ciprofloxacin but not to other agents and contained a mutation resulting in the replacement of threonine 86 with isoleucine in the quinolone resistance-determining region of GyrA. Mutants with two distinct phenotypes were selected from the efflux pump knockout strains. Mutants with the first phenotype were resistant to ciprofloxacin only and had the same substitution within GyrA as the NCTC11168-derived mutants. Irrespective of the parent strain, mutants with the second phenotype were resistant to ciprofloxacin, chloramphenicol, tetracycline, ethidium bromide, acridine orange, and SDS and had no mutation in gyrA. These mutants expressed levels of the efflux pump genes cmeB and cmeF and the major outer membrane protein gene porA similar to those expressed by the respective parent strains. No mutations were detected in cmeF or cmeB. Accumulation assays revealed that the mutants accumulated lower concentrations of drug. These data suggest the involvement of a non-CmeB or -CmeF efflux pump or reduced uptake conferring multiple-antibiotic resistance, which can be selected after exposure to a fluoroquinolone. PMID- 15793097 TI - Molecular detection of the macrolide efflux gene: to discriminate or not to discriminate between mef(A) and mef(E). PMID- 15793100 TI - In vitro susceptibility testing of Borrelia burgdorferi sensu lato isolates cultured from patients with erythema migrans before and after antimicrobial chemotherapy. AB - Clinical treatment failures have been reported to occur in early Lyme borreliosis (LB) for many suitable antimicrobial agents. Investigations of possible resistance mechanisms of the Borrelia burgdorferi complex must analyze clinical isolates obtained from LB patients, despite their receiving antibiotic treatment. Here, borrelial isolates obtained from five patients with erythema migrans (EM) before the start of antibiotic therapy and again after the conclusion of treatment were investigated. The 10 isolates were characterized by restriction fragment length polymorphism analysis and plasmid profile analysis and subjected to susceptibility testing against a variety of antimicrobial agents including those used for initial chemotherapy. Four out of five patients were infected by the same genospecies (Borrelia afzelii, n = 3; Borrelia garinii, n = 1) at the site of the EM lesion before and after antimicrobial therapy. In one patient the genospecies of the initial isolate (B. afzelii) differed from that of the follow up isolate (B. garinii). No significant changes in the in vitro susceptibilities became obvious for corresponding clinical isolates before the start and after the conclusion of antimicrobial therapy. This holds true for the antimicrobial agents used for specific chemotherapy of the patients, as well as for any of the additional agents tested in vitro. Our study substantiates borrelial persistence in some EM patients at the site of the infectious lesion despite antibiotic treatment over a reasonable time period. Borrelial persistence, however, was not caused by increasing MICs or minimal borreliacidal concentrations in these isolates. Therefore, resistance mechanisms other than acquired resistance to antimicrobial agents should be considered in patients with LB resistant to treatment. PMID- 15793101 TI - Glycerol monolaurate inhibits virulence factor production in Bacillus anthracis. AB - Anthrax, caused by Bacillus anthracis, has been brought to the public's attention because of the 2001 bioterrorism attacks. However, anthrax is a disease that poses agricultural threats in the United States as well as human populations in Europe, China, Africa, and Australia. Glycerol monolaurate (GML) is a compound that has been shown to inhibit exotoxin production by Staphylococcus aureus and other gram-positive bacteria. Here, we study the effects of GML on growth and toxin production in B. anthracis. The Sterne strain of B. anthracis was grown to post-exponential phase with 0-, 10-, 15-, or 20-microg/ml concentrations of GML and then assayed quantitatively for protective antigen (PA) and lethal factor (LF). After 8 h, GML at concentrations greater than 20 microg/ml was bacteriostatic to growth of the organism. However, a 10-microg/ml concentration of GML was not growth inhibitory, but amounts of PA and LF made were greatly reduced. This effect was not global for all proteins when total secreted protein from culture fluids was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Through quantitative reverse transcription-PCR assays, this toxin-inhibitory effect was shown to occur at the transcriptional level, since amounts of mRNA for pagA (PA), lef (LF), and cya (edema factor) were reduced. Surprisingly, mRNA levels of atxA, a regulator of exotoxin gene expression, rose in the presence of GML. These data will be useful in developing therapeutic tools to treat anthrax disease, whether in animals or humans. These results also suggest that mechanisms of virulence regulation exist independent of atxA. PMID- 15793102 TI - Pseudomonas aeruginosa bloodstream infection: importance of appropriate initial antimicrobial treatment. AB - Pseudomonas aeruginosa bloodstream infection is a serious infection with significant patient mortality and health-care costs. Nevertheless, the relationship between initial appropriate antimicrobial treatment and clinical outcomes is not well established. This study was a retrospective cohort analysis employing automated patient medical records and the pharmacy database at Barnes Jewish Hospital. Three hundred five patients with P. aeruginosa bloodstream infection were identified over a 6-year period (January 1997 through December 2002). Sixty-four (21.0%) patients died during hospitalization. Hospital mortality was statistically greater for patients receiving inappropriate initial antimicrobial treatment (n = 75) compared to appropriate initial treatment (n = 230) (30.7% versus 17.8%; P = 0.018). Multiple logistic regression analysis identified inappropriate initial antimicrobial treatment (adjusted odds ratio [AOR], 2.04; 95% confidence interval [CI], 1.42 to 2.92; P = 0.048), respiratory failure (AOR, 5.18; 95% CI, 3.30 to 8.13; P < 0.001), and circulatory shock (AOR, 4.00; 95% CI, 2.71 to 5.91; P < 0.001) as independent determinants of hospital mortality. Appropriate initial antimicrobial treatment was administered statistically more often among patients receiving empirical combination antimicrobial treatment for gram-negative bacteria compared to empirical monotherapy (79.4% versus 65.5%; P = 0.011). Inappropriate initial empirical antimicrobial treatment is associated with greater hospital mortality among patients with P. aeruginosa bloodstream infection. Inappropriate antimicrobial treatment of P. aeruginosa bloodstream infections may be minimized by increased use of combination antimicrobial treatment until susceptibility results become known. PMID- 15793104 TI - bla(CTX-M) genes in clinical Salmonella isolates recovered from humans in England and Wales from 1992 to 2003. AB - Cefotaximases (CTX-M) are a rapidly growing class A beta-lactamase family that has been found among a wide range of clinical bacteria. One hundred and six isolates were selected from 278,308 Salmonella isolates based on resistance to ampicillin and cephalosporins and subjected to further characterization. Fourteen isolates were bla(CTX-M) PCR positive, and cefotaxime MICs for these isolates were > or = 16 mg/liter. Furthermore, sequence analysis revealed the presence of type CTX-M9, -15, or -17 to -18. All 14 isolates presented different PFGE restriction profiles, although six Salmonella enterica serotype Virchow isolates formed a tight cluster. The bla(CTX-M) genetic determinants were present in transferable plasmids of approximately 63, 105, and >148 kb. Plasmid restriction analysis showed that both horizontal transfer of similar plasmids among different clones and transfer of genes between different plasmids were likely mechanisms involved in the spread of bla(CTX-M) genes. We have found that CTX-M enzymes have emerged in community-acquired infections both linked to foreign travel and domestically acquired. This is the first report of a CTX-M enzyme in Salmonella in the United Kingdom. Also, it represents the first report of a bla(CTX-M) gene in Salmonella enterica serotype Stanley and a bla(CTX-M-15) gene in Salmonella enterica serotypes Anatum, Enteritidis, and Typhimurium. PMID- 15793103 TI - Reduced azole susceptibility in genotype 3 Candida dubliniensis isolates associated with increased CdCDR1 and CdCDR2 expression. AB - Candida dubliniensis is a recently identified yeast species primarily associated with oral carriage and infection in individuals infected with the human immunodeficiency virus. The species can be divided into at least four genotypes on the basis of the nucleotide sequence of the internal transcribed spacer region of the rRNA operon. Previous studies have shown that a small number of clinical isolates belonging to genotype 1 are resistant to the commonly used antifungal drug fluconazole. The aim of the present study was to investigate the molecular mechanisms responsible for reduced susceptibility to azole drugs in C. dubliniensis genotype 3 isolates obtained from a patient with fluconazole recalcitrant oral candidiasis. Four isolates from a single clinical sample, one susceptible, the other three exhibiting reduced susceptibilities to fluconazole, itraconazole, ketoconazole, voriconazole, and posaconazole, were examined. Results showed that reduced susceptibility to azole drugs was associated with an increase in the expression of the multidrug transporters CdCDR1 and CdCDR2 which correlated with reduced intracellular accumulation of radiolabeled fluconazole and an increase in the activity of energy-dependent efflux mechanisms. In contrast to observations made in previous studies, overexpression of the multidrug transporter CdMDR1 was not observed. Despite a thorough investigation of all commonly encountered mechanisms of azole resistance, no other mechanism could be associated with reduced susceptibility to azole drugs in the clinical isolates studied. This is the first report of CdCDR2 involvement in azole resistance in C. dubliniensis. PMID- 15793105 TI - Is in vitro antibiotic combination more effective than single-drug therapy against anthrax? AB - Antibiotic combinations are used to enhance antibacterial efficacy and to prevent the development of resistance. We have tested a possible synergistic effect of several antibacterial combinations on Bacillus anthracis. The in vitro activities of antibiotic combinations against two strains of B. anthracis, strain Sterne and the Russian anthrax vaccine strain STi, were tested by the fractional inhibitory concentration (FIC) method, derived from the MICs of the agents in combination, and by measuring the rate of bacterial killing over time by several antibiotic combinations. The FIC results showed that synergism against both B. anthracis strains was observed only with the combination of rifampin and clindamycin. The telithromycin-amoxicillin combination showed synergism against strain Sterne only. All other combinations were either indifferent or antagonistic. The results of the bacterial time-kill study demonstrated indifferent effects for all combinations. These in vitro results demonstrate the difficulties in obtaining synergistic combinations of antibiotics against B. anthracis. PMID- 15793106 TI - Activity of Hoechst 33258 against Pneumocystis carinii f. sp. muris, Candida albicans, and Candida dubliniensis. AB - Hoechst 33258 is a compound that binds nucleic acids. We report that Hoechst 33258 exhibits antimicrobial activity against Pneumocystis carinii f. sp. muris in a mouse model for P. carinii pneumonia and against Candida albicans and Candida dubliniensis in vitro. Relative to saline treatment, a 14-day, daily treatment of mice with 37.5 mg of Hoechst 33258/kg of body weight after inoculation with P. carinii reduced by about 100-fold the number of P. carinii organisms detected by either PCR or by microscopy after silver staining. For comparison, treatment based on a dose of 15 to 20 mg of the trimethoprim component in trimethoprim-sulfamethoxazole/kg reduced the number of P. carinii by about fourfold. In vitro inhibition of P. carinii group I intron splicing was observed with a 50% inhibitory concentration (IC50) of 30 microM in 2 or 4 mM Mg2+, suggesting RNA as a possible target. However, Hoechst 33258 inhibits growth of Candida strains with and without group I introns. IC50s ranged from 1 to 9 microM for strains with group I introns and were 12 and 32 microM for two strains without group I introns. These studies demonstrate that compounds that bind fungal nucleic acids have the potential to be developed as new therapeutics for Pneumocystis and possibly other fungi, especially if they could be directed to structures that are not present in mammalian cells, such as self-splicing introns. PMID- 15793107 TI - Pharmacokinetic and maximum tolerated dose study of micafungin in combination with fluconazole versus fluconazole alone for prophylaxis of fungal infections in adult patients undergoing a bone marrow or peripheral stem cell transplant. AB - In this dose escalation study, 74 adult cancer patients undergoing bone marrow or peripheral blood stem cell transplantation received fluconazole (400 mg/day) and either normal saline (control) (12 subjects) or micafungin (12.5 to 200 mg/day) (62 subjects) for up to 4 weeks. The maximum tolerated dose (MTD) of micafungin was not reached, based on the development of Southwest Oncology Group criteria for grade 3 toxicity; drug-related toxicities were rare. Commonly occurring adverse events considered related to micafungin were headache (6.8%), arthralgia (6.8%), hypophosphatemia (4.1%), insomnia (4.1%), maculopapular rash (4.1%), and rash (4.1%). Pharmacokinetic profiles for micafungin on days 1 and 7 were similar. The mean half-life was approximately 13 h, with little variance after repeated or increasing doses. Mean maximum concentrations of the drug in serum and areas under the concentration-time curve from 0 to 24 h were approximately proportional to dose. There was no clinical or kinetic evidence of interaction between micafungin and fluconazole. Five of 12 patients (42%) in the control group and 14 of 62 (23%) in the micafungin-plus-fluconazole groups had a suspected fungal infection during treatment which resulted in empirical treatment with amphotericin B. The combination of micafungin and fluconazole was found to be safe in this high-risk patient population. The MTD of micafungin was not reached even at doses up to 200 mg/day for 4 weeks. The pharmacokinetic profile of micafungin in adult cancer patients with blood or marrow transplants is consistent with the profile in healthy volunteers, and the area under the curve is proportional to dose. PMID- 15793108 TI - Comparison of the pharmacodynamics of meropenem in patients with ventilator associated pneumonia following administration by 3-hour infusion or bolus injection. AB - The time that concentrations in serum are above the MIC (T>MIC) is the pharmacokinetic/pharmacodynamic parameter correlating with the therapeutic efficacy of beta-lactam antibiotics. The aim of this study was to demonstrate the T>MIC of meropenem when administered by a 3-h infusion compared with that when administered by bolus injection. The study was conducted with nine patients with ventilator-associated pneumonia. Each subject received meropenem in three regimens consecutively: (i) bolus injection of 1 g every 8 h for 24 h; (ii) 3-h infusion of 1 g every 8 h for 24 h; and (iii) 3-h infusion of 2 g every 8 h for 24 h. Following bolus injection, the percentages of the T>MICs of 16, 8, 4, and 1 microg/ml were 28.33% +/- 11.67%, 45.89% +/- 22.90%, 57.00% +/- 24.82%, and 74.67% +/- 17.94% of an 8-h interval, respectively. For the 3-h infusion of 1 g of meropenem, the percentages of the T>MICs of 16, 8, 4, and 1 microg/ml were 37.78% +/- 20.57%, 58.11% +/- 24.38%, 72.67% +/- 21.97%, and 93.56% +/- 6.84% of an 8-h interval, respectively. For the 3-h infusion of 2 g of meropenem, the percentages of the T>MICs of 16, 8, 4, and 1 microg/ml were 57.89% +/- 24.26%, 72.89% +/- 22.40%, 85.56% +/- 16.42%, and 98.56% +/- 3.28% of an 8-h interval, respectively. In conclusion, a 3-h infusion resulted in greater T>MICs than those after a bolus injection. For the treatment of infections caused by pathogens with intermediate resistance, a 3-h infusion of 2 g of meropenem every 8 h can provide concentrations in serum above the MIC of 16 microg/ml for almost 60% of an 8-h interval. PMID- 15793109 TI - Bactericidal and antiendotoxic properties of short cationic peptides derived from a snake venom Lys49 phospholipase A2. AB - The activities of short synthetic, nonhemolytic peptides derived from the C terminal region of myotoxin II, a catalytically inactive phospholipase A2 homologue present in the venom of the snake Bothrops asper, have been shown to reproduce the bactericidal activity of the parent protein. They combine cationic and hydrophobic-aromatic amino acids, thus functionally resembling the antimicrobial peptides of innate defenses. This study evaluated the antimicrobial and antiendotoxic properties of a 13-mer derivative peptide of the C-terminal sequence from positions 115 to 129 of myotoxin II, named pEM-2. This peptide (KKWRWWLKALAKK) showed bactericidal activity against both gram-positive and gram negative bacteria. In comparison to previously described peptide variants derived from myotoxin II, the toxicity of pEM-2 toward eukaryotic cells in culture was significantly reduced, being similar to that of lactoferricin B but lower than that of polymyxin B. The all-D enantiomer of pEM-2 [pEM-2 (D)] retained the same bactericidal potency of its L-enantiomeric counterpart, but it showed an enhanced ability to counteract the lethal activity of an intraperitoneal lipopolysaccharide challenge in mice, which correlated with a significant reduction of the serum tumor necrosis factor alpha levels triggered by this endotoxin. Lethality induced by intraperitoneal infection of mice with Escherichia coli or Salmonella enterica serovar Typhimurium was reduced by the administration of pEM-2 (D). These results demonstrate that phospholipase A2 derived peptides may have the potential to counteract microbial infections and encourage further evaluations of their actions in vivo. PMID- 15793111 TI - Successive emergence of Enterobacter aerogenes strains resistant to imipenem and colistin in a patient. AB - Enterobacter aerogenes is an agent of hospital-acquired infection that exhibits a remarkable resistance to beta-lactam antibiotics during therapy. Five successive isolates of E. aerogenes infecting a patient and exhibiting a multiresistance phenotype to beta-lactam antibiotics and fluoroquinolones were investigated. Among these clinical strains, four presented resistant phenotypes during successive imipenem and colistin treatments. The involved resistance mechanisms exhibited by the successive isolates were associated with alterations of the outer membrane that caused a porin decrease and lipopolysaccharide modifications. PMID- 15793110 TI - Development of a cell-based high-throughput specificity screen using a hepatitis C virus-bovine viral diarrhea virus dual replicon assay. AB - The hepatitis C virus (HCV) replicon is a unique system for the development of a high-throughput screen (HTS), since the analysis of inhibitors requires the quantification of a decrease in a steady-state level of HCV RNA. HCV replicon replication is dependent on host cell factors, and any toxic effects may have a significant impact on HCV replicon replication. Therefore, determining the antiviral specificity of compounds presents a challenge for the identification of specific HCV inhibitors. Here we report the development of an HCV/bovine viral diarrhea virus (BVDV) dual replicon assay suitable for HTS to address these issues. The HCV reporter enzyme is the endogenous NS3 protease contained within the HCV genome, while the BVDV reporter enzyme is a luciferase enzyme engineered into the BVDV genome. The HTS uses a mixture of HCV and BVDV replicon cell lines placed in the same well of a 96-well plate and isolated in the same cell backgrounds (Huh-7). The format consists of three separate but compatible assays: the first quantitates the amount of cytotoxicity based upon the conversion of Alamar blue dye via cellular enzymes, while the second indirectly quantitates HCV replicon replication through measurement of the amount of NS3 protease activity present. The final assay measures the amount of luciferase activity present from the BVDV replicon cells, as an indicator of the specificity of the test compounds. This HCV/BVDV dual replicon assay provides a reliable format to determine the potency and specificity of HCV replicon inhibitors. PMID- 15793112 TI - Topical caspofungin for treatment of keratitis caused by Candida albicans in a rabbit model. AB - Candida albicans is the most frequent cause of fungal keratitis in temperate regions. Caspofungin has potent activity against Candida spp. in a variety of clinical settings. Little is known, however, about its activity against fungal keratitis. We compared the efficacy of topical caspofungin with that of topical amphotericin B (AMB) in a rabbit model of experimental keratomycosis. Keratitis was induced with a standardized inoculum of Candida albicans (SC 5314) placed on the debrided cornea. Twenty-four hours after infection, animals were randomly assigned to treatment with 0.15% caspofungin, 0.5% caspofungin, 0.15% AMB, and a saline control (n = 12 rabbits in each group). For the first 12 h, treatment was repeated every 30 min and, after a 12-h pause, was resumed at hourly intervals for another 12 h. The animals were examined and killed 12 h after administration of the last dose. Treatment effects were evaluated by clinical assessment, fungal culture, and histopathology. Drug treatment significantly reduced corneal fungal recovery from 3.78 log10 CFU in saline-treated animals to 2.97, 1.76, and 1.18 log10 CFU in animals treated with 0.15% caspofungin, 0.5% caspofungin, and 0.15% AMB, respectively. By histopathology, the mean hyphal density was significantly lower in the corneas of treated animals than in those of the controls; there was no difference in hyphal densities between the different treatment groups. The depth of corneal invasion was not significantly reduced by the antifungal treatments. By clinical assessment, keratitis progressed in animals treated with saline, whereas disease progression was inhibited by all drug treatment regimens. In our rabbit model, 0.5% caspofungin was as effective as 0.15% AMB for the topical treatment of Candida keratitis. The potential clinical efficacy of caspofungin awaits further investigation. PMID- 15793113 TI - Testing of the in vitro susceptibilities of Madurella mycetomatis to six antifungal agents by using the Sensititre system in comparison with a viability based 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5- [(phenylamino)carbonyl]-2H tetrazolium hydroxide (XTT) assay and a modified NCCLS method. AB - The in vitro susceptibilities of 36 clinical isolates of Madurella mycetomatis, the prime agent of eumycetoma in Africa, to ketoconazole, itraconazole, fluconazole, voriconazole, amphotericin B, and flucytosine were determined by the Sensititre YeastOne system. This system appeared to be a rapid and easy test, and by use of hyphal suspensions it generated results comparable to those of a modified NCCLS method. After 10 days of incubation, the antifungal activities of ketoconazole (MIC at which 90% of isolates were inhibited [MIC90], 0.125 microg/ml), itraconazole (MIC90, 0.064 microg/ml), and voriconazole (MIC90, 0.125 microg/ml) appeared superior to those of fluconazole (MIC90, 128 microg/ml) and amphotericin B (MIC90, 1 microg/ml), with MICs in the clinically relevant range. All isolates were resistant to flucytosine (all MICs above 64 microg/ml). Based on the relatively broad range of MICs obtained for the antifungal agents, routine testing of M. mycetomatis isolates for susceptibility to antifungal agents seems to be relevant to adequate therapeutic management. PMID- 15793114 TI - Efficacy of caspofungin against central nervous system Aspergillus fumigatus infection in mice determined by TaqMan PCR and CFU methods. AB - We have reported previously that prolonged caspofungin (CAS) dosing enhances survival in a murine model of central nervous system aspergillosis. In this study we determined by quantitative PCR (qPCR) and CFU enumeration whether CAS could reduce fungal burdens, prior to the deaths of untreated animals, and also assessed progressive infection in untreated mice. Mice were infected intracranially and treated for 4 days with CAS (1, 5, or 10 mg/kg of body weight/day) or amphotericin B (AMB) (3 mg/kg/day) starting 1 day postinfection. Fungal burdens in brains and kidneys of untreated controls were determined on days 1, 3, and 5 to assess progressive infection; burdens in treated animals were determined on day 5. qPCR showed higher burdens than CFU enumeration in all comparisons. In untreated animals, qPCR showed transiently increased burdens in brains, while CFU enumeration showed a decrease. qPCR showed increased burdens in kidneys, but CFU enumeration did not. Neither method indicated drug efficacy in the brain. Both methods showed AMB efficacy in the kidneys, and qPCR demonstrated CAS efficacy at all doses. Spearman correlations of qPCR and CFU determination results showed a significant correlation for most untreated groups; results correlated well for kidneys (P < or = 0.03) but not for brains in treated mice. Regression analyses of qPCR and CFU groups indicated different slopes for progressive infection in untreated animals but the same slopes for CAS dose response efficacy. qPCR appeared to better reflect the progression of untreated infection. The lack of demonstration of efficacy in the brain suggests that longer dosing is necessary to cause burden reduction. These results also suggest that, when there is drug efficacy in a therapeutic study, either method appears to be useful for determining Aspergillus fumigatus burdens. PMID- 15793115 TI - Azithromycin exhibits bactericidal effects on Pseudomonas aeruginosa through interaction with the outer membrane. AB - The aim of the present study was to elucidate the effect of the macrolide antibiotic azithromycin on Pseudomonas aeruginosa. We studied the susceptibility to azithromycin in P. aeruginosa PAO1 using a killing assay. PAO1 cells at the exponential growth phase were resistant to azithromycin. In contrast, PAO1 cells at the stationary growth phase were sensitive to azithromycin. The divalent cations Mg2+ and Ca2+ inhibited this activity, suggesting that the action of azithromycin is mediated by interaction with the outer membranes of the cells, since the divalent cations exist between adjacent lipopolysaccharides (LPSs) and stabilize the outer membrane. The divalent cation chelator EDTA behaved in a manner resembling that of azithromycin; EDTA killed more PAO1 in the stationary growth phase than in the exponential growth phase. A 1-N-phenylnaphthylamine assay showed that azithromycin interacted with the outer membrane of P. aeruginosa PAO1 and increased its permeability while Mg2+ and Ca2+ antagonized this action. Our results indicate that azithromycin directly interacts with the outer membrane of P. aeruginosa PAO1 by displacement of divalent cations from their binding sites on LPS. This action explains, at least in part, the effectiveness of sub-MICs of macrolide antibiotics in pseudomonal chronic airway infection. PMID- 15793116 TI - Development of cell-based assays for in vitro characterization of hepatitis C virus NS3/4A protease inhibitors. AB - A recombinant vaccinia virus, expressing the NS3-to-NS5 region of the N clone of hepatitis C virus (HCV), was generated and utilized both in a gel-based assay and in an enzyme-linked immunosorbent assay (ELISA) to evaluate the pyrrolidine-5,5 trans-lactams, a series of inhibitors of the HCV NS3/4A protease. The absolute levels of processed, mature HCV nonstructural proteins in this system were found to decrease in the presence of the trans-lactams. Monitoring of this reduction enabled end points and 50% inhibitory concentrations to be calculated in order to rank the active compounds according to potency. These compounds had no effect on the transcription or translation of the NS3-5 polyprotein at concentrations shown to inhibit NS3/4A protease, and they were shown to be specific inhibitors of this protease. The ELISA, originally developed using the vaccinia virus expression system, was modified to utilize Huh-7 cells containing an HCV replicon. Results with this assay correlated well with those obtained with the recombinant vaccinia virus assays. These results demonstrate the utility of these assays for the characterization of NS3/4A protease inhibitors. In addition, inhibitors of other viral targets, such as polymerase and helicase, can be evaluated in the context of the replicon ELISA. PMID- 15793117 TI - Phototargeting oral black-pigmented bacteria. AB - We have found that broadband light (380 to 520 nm) rapidly and selectively kills oral black-pigmented bacteria (BPB) in pure cultures and in dental plaque samples obtained from human subjects with chronic periodontitis. We hypothesize that this killing effect is a result of light excitation of their endogenous porphyrins. Cultures of Prevotella intermedia and P. nigrescens were killed by 4.2 J/cm2, whereas P. melaninogenica required 21 J/cm2. Exposure to light with a fluence of 42 J/cm2 produced 99% killing of P. gingivalis. High-performance liquid chromatography demonstrated the presence of various amounts of different porphyrin molecules in BPB. The amounts of endogenous porphyrin in BPB were 267 (P. intermedia), 47 (P. nigrescens), 41 (P. melaninogenica), and 2.2 (P. gingivalis) ng/mg. Analysis of bacteria in dental plaque samples by DNA-DNA hybridization for 40 taxa before and after phototherapy showed that the growth of the four BPB was decreased by 2 and 3 times after irradiation at energy fluences of 4.2 and 21 J/cm2, respectively, whereas the growth of the remaining 36 microorganisms was decreased by 1.5 times at both energy fluences. The present study suggests that intraoral light exposure may be used to control BPB growth and possibly benefit patients with periodontal disease. PMID- 15793118 TI - Differential expression of cytokines and chemokines in human monocytes induced by lipid formulations of amphotericin B. AB - The immunomodulatory effects of liposomal amphotericin B (LAMB), amphotericin B lipid complex (ABLC), and amphotericin B colloidal dispersion (ABCD) on mRNA and protein profiles of five cytokines and chemokines expressed by human monocyte enriched mononuclear leukocytes (MNCs) were comprehensively evaluated by semiquantitative reverse transcription-PCR and enzyme-linked immunosorbent assays; they were compared to those of deoxycholate amphotericin B (DAMB). mRNAs of interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL-1ra), tumor necrosis factor alpha (TNF-alpha), monocyte chemotactic protein 1 (MCP-1), and macrophage inflammatory protein 1beta (MIP-1beta) were assessed after treatment of MNCs with each drug for 0.5, 2, 6, and 22 h. The cytokine protein profiles were obtained after incubation of MNCs with the drugs for 2 h (TNF-alpha) or 6 h (all the others). In the mRNA studies, DAMB resulted in an early increase of inflammatory cytokines or chemokines IL-1beta, TNF-alpha, MCP-1, and MIP-1beta (2 to 6 h) and in a late increase of anti-inflammatory IL-1ra (22 h). ABCD showed a general similar trend of inflammatory gene up-regulation. LAMB and ABLC decreased or did not affect IL-1beta and TNF-alpha, whereas ABLC additionally decreased MIP 1beta. In protein measurement studies, DAMB and ABCD up-regulated production of IL-1beta (P < 0.05), decreased the IL-1ra/IL-1beta ratio, and up-regulated the production of MCP-1 and MIP-1beta. In comparison, LAMB and ABLC down-regulated or did not affect the production of these cytokines/chemokines compared to untreated MNCs; furthermore, ABLC tended to increase the IL-1ra/IL-1beta ratio. These studies demonstrate that amphotericin B formulations differentially affect gene expression and release of an array of proinflammatory and anti-inflammatory cytokines that potentially may explain the differences in infusion-related reactions and dose-dependent nephrotoxicity as well as modulation of the host immune response to invasive fungal infections. PMID- 15793119 TI - Susceptibility to antibiotics and beta-lactamase induction in murein hydrolase mutants of Escherichia coli. AB - The antibiotic susceptibilities and capabilities to induce beta-lactamases were studied in multiple Escherichia coli murein (peptidoglycan) hydrolase mutants. E. coli mutants lacking either three amidases, three amidases and one lytic transglycosylase, or six lytic transglycosylases showed higher levels of susceptibility to bacitracin, erythromycin, gallidermin, and vancomycin than the wild type. Mutant cells without three amidases lost viability in the presence of vancomycin and gallidermin, whereas the wild type was resistant to both antibiotics. Beta-lactamase induction was studied after introduction of a plasmid carrying the ampC and ampR genes. Upon addition of cefoxitin to the growth medium, the wild type as well as a mutant lacking all known amidases and DD endopeptidases induced beta-lactamase, whereas a mutant lacking all known lytic transglycosylases was unable to induce beta-lactamase, showing that lytic transglycosylase activity is essential for beta-lactamase induction. Consequently, cells lacking lytic transglycosylase activity lysed in the presence of penicillin, despite the presence of the inducible beta-lactamase system. We discuss the potential of murein hydrolase inhibitors for antibiotic therapy. PMID- 15793120 TI - Scintillation proximity assay for inhibitors of Escherichia coli MurG and, optionally, MraY. AB - MurG and MraY, essential enzymes involved in the synthesis of bacterial peptidoglycan, are difficult to assay because the substrates are lipidic and hard to prepare in large quantities. Based on the use of Escherichia coli membranes lacking PBP1b, we report a high-throughput method to measure the activity of MurG and, optionally, MraY as well. In these membranes, incubation with the two peptidoglycan sugar precursors results in accumulation of lipid II rather than the peptidoglycan produced by wild-type membranes. MurG was assayed by addition of UDP-[3H]N-acetylglucosamine to membranes in which lipid I was preformed by incubation with UDP-N-acetyl-muramylpentapeptide, and the product was captured by wheat germ agglutinin scintillation proximity assay beads. In a modification of the assay, the activity of MraY was coupled to that of MurG by addition of both sugar precursors together in a single step. This allows simultaneous detection of inhibitors of either enzyme. Both assays could be performed using wild-type membranes by addition of the transglycosylase inhibitor moenomycin. Nisin and vancomycin inhibited the MurG reaction; the MraY-MurG assay was inhibited by tunicamycin as well. Inhibitors of other enzymes of peptidoglycan synthesis- penicillin G, moenomycin, and bacitracin--had no effect. Surprisingly, however, the beta-lactam cephalosporin C inhibited both the MurG and MraY-MurG assays, indicating a secondary mechanism by which this drug inhibits bacterial growth. In addition, it inhibited NADH dehydrogenase in membranes, a hitherto-unreported activity. These assays can be used to screen for novel antibacterial agents. PMID- 15793121 TI - Silencing of glycopeptide resistance in Enterococcus faecalis BM4405 by novobiocin. AB - Enterococcus faecalis BM4405-1, a susceptible derivative of the VanE-type vancomycin-resistant E. faecalis strain BM4405, was obtained after growth in the presence of novobiocin, an inhibitor of the GyrB subunit of DNA gyrase. In contrast to findings for BM4405, UDP-MurNAc-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala (pentapeptide[D-Ala]) was the only peptidoglycan precursor found in BM4405-1, and no VanXY(E) D,D-peptidase or VanT serine racemase activities were detected in that strain, even after induction by subinhibitory concentrations of vancomycin. Sequencing of the vanE operon of BM4405-1 revealed two mutations leading to substitutions in VanE (D200N) and in the C-terminal amino acid of VanR(E) (Y225F). Cloning of the vanE, vanXY(E), and vanT(E) genes of BM4405-1 into the susceptible E. faecalis strain JH2-2 conferred resistance to vancomycin, indicating that the mutation in vanE was not responsible for susceptibility. Transcriptional analysis of the vanE operon in BM4405 by quantitative reverse transcription-PCR indicated that novobiocin did not affect the expression level of the vanE operon. Sequencing of the gyrB gene of BM4405-1 revealed a mutation responsible for substitution of a residue (K337Y) required for ATPase activity and thus implicated in DNA supercoiling. Cloning of the gyrB gene of BM4405 restored vancomycin resistance to BM4405-1. Taken together, these data suggest that alteration of DNA supercoiling following a mutation in GyrB was responsible for lack of expression of the vanE operon and thus for vancomycin susceptibility in BM4405-1. PMID- 15793122 TI - Compensatory adaptation to the loss of biological fitness associated with acquisition of fusidic acid resistance in Staphylococcus aureus. AB - Recent studies have shown that individual amino acid exchanges within elongation factor G (EF-G) cause fusidic acid resistance in Staphylococcus aureus. The data from the present study illustrate that the fusidic acid resistance-mediating amino acid substitutions P406L and H457Y are associated with a marked impairment of the biological fitness of S. aureus. In particular, strains producing EF-G derivatives with these mutations showed reduced growth, decreased plasma coagulase activity, and an impaired capability to compete with the isogenic wild type strain. Second-site mutations within EF-G, such as A67T and S416F, that have been encountered in clinical fusidic acid-resistant isolates containing the amino acid exchanges P406L and H457Y, respectively, were shown not to contribute to resistance. Furthermore, the substitution A67T had no impact on the biological fitness in vitro. The exchange S416F, however, was found to function as a fitness compensating mutation in S. aureus carrying the substitution H457Y in EF-G. In conclusion, the data presented in this report provide evidence at the molecular level that the deleterious effects of fusidic acid resistance-mediating exchanges within EF-G of S. aureus can be reduced considerably by specific compensating mutations in this target protein. This compensatory adaptation most likely plays a significant role in the stabilization of resistant bacteria within a given population. PMID- 15793123 TI - Acquisition of resistance to carbapenems in multidrug-resistant clinical strains of Acinetobacter baumannii: natural insertional inactivation of a gene encoding a member of a novel family of beta-barrel outer membrane proteins. AB - The outer membrane proteins responsible for the influx of carbapenem beta-lactam antibiotics in the nonfermentative gram-negative pathogen Acinetobacter baumannii are still poorly characterized. Resistance to both imipenem and meropenem in multidrug-resistant clinical strains of A. baumannii is associated with the loss of a heat-modifiable 29-kDa outer membrane protein, designated CarO. The chromosomal locus containing the carO gene was cloned and characterized from different clinical isolates. Only one carO copy, present in a single transcriptional unit, was found in the A. baumannii genome. The carO gene encodes a polypeptide of 247 amino acid residues with a typical N-terminal signal sequence and a predicted transmembrane beta-barrel topology. Its absence from different carbapenem-resistant clinical isolates of A. baumannii resulted from the disruption of carO by distinct insertion elements. The overall data thus support the notion that CarO participates in the influx of carbapenem antibiotics in A. baumannii. Moreover, database searches identified the presence of carO homologs only in species of the genera Acinetobacter, Moraxella, and Psychrobacter, disclosing the existence of a novel family of outer membrane proteins restricted to the family Moraxellaceae of the class gamma Proteobacteria. PMID- 15793124 TI - Topoisomerase I amino acid substitutions, Gly185Arg and Asp325Glu, confer camptothecin resistance in Leishmania donovani. AB - The antitumor compound camptothecin (CPT) is also recognized for its specific activity against Leishmania donovani topoisomerase I (Topo-I). In consequence, defining CPT resistance mechanisms represents an important strategic tool in the acquisition of a better understanding of its mode of action. In the present study, we selected a single highly resistant L. donovani strain termed LdRCPT.160 by stepwise exposure to CPT. Gene sequencing revealed two single nucleotide mutations in the LdRCPT.160 LdTOP1A gene, resulting in two amino acid substitutions (Gly185Arg and Asp325Glu) in the protein. Moreover, these two substitutions observed in the LdTOP1A protein were correlated with a decreased Topo-I DNA relaxation activity in these resistant parasites. Nevertheless, there was no change in the LdTOP1A gene expression level. Interestingly, transfection studies of the LdRCPT.160 LdTOP1A gene in its wild-type counterpart showed that it induced CPT resistance. Site-directed mutagenesis studies demonstrated that, despite a substantial level of resistance conferred by the Gly185Arg and Asp325Glu substitutions separately, both were essential to reach a high resistance phenotype. Of interest, the amino acid substitutions observed in LdRCPT.160 LdTOP1A protein occurred near the amino acids previously predicted to interact with CPT, providing new insight into the mechanism of CPT molecular action. PMID- 15793125 TI - In vitro and in vivo activities of macrolide derivatives against Mycobacterium tuberculosis. AB - Existing macrolides have never shown definitive clinical efficacy in tuberculosis. Recent reports suggest that ribosome methylation is involved in macrolide resistance in Mycobacterium tuberculosis, a mechanism that newer macrolides have been designed to overcome in gram-positive bacteria. Therefore, selected macrolides and ketolides (descladinose) with substitutions at positions 9, 11,12, and 6 were assessed for activity against M. tuberculosis, and those with MICs of < or = 4 microM were evaluated for cytotoxicity to Vero cells and J774A.1 macrophages. Several compounds with 9-oxime substitutions or aryl substitutions at position 6 or on 11,12 carbamates or carbazates demonstrated submicromolar MICs. For the three macrolide-ketolide pairs, macrolides demonstrated superior activity. Four compounds with low MICs and low cytotoxicity also effected significant reductions in CFU in infected macrophages. Active compounds were assessed for tolerance and the ability to reduce CFU in the lungs of BALB/c mice in an aerosol infection model. A substituted 11,12 carbazate macrolide demonstrated significant dose-dependent inhibition of M. tuberculosis growth in mice, with a 10- to 20-fold reduction of CFU in lung tissue. Structure activity relationships, some of which are unique to M. tuberculosis, suggest several synthetic directions for further improvement of antituberculosis activity. This class appears promising for yielding a clinically useful agent for tuberculosis. PMID- 15793126 TI - Molecular analysis of isoniazid-resistant Mycobacterium tuberculosis isolates from England and Wales reveals the phylogenetic significance of the ahpC -46A polymorphism. AB - The present study investigated the prevalence and diagnostic potential of the most commonly reported mutations associated with isoniazid resistance, katG 315Thr, katG 315Asn, inhA -15T, inhA -8A, and the oxyR-ahpC intergenic region, in a population sample of 202 isoniazid-resistant Mycobacterium tuberculosis isolates and 176 randomly selected fully sensitive isolates from England and Wales identified by using a directed oligonucleotide array and limited DNA sequencing. The strains were recovered from patients originating from 29 countries; 41 isolates were multidrug resistant. Mutations affecting katG 315, the inhA promoter, and the oxyR-ahpC intergenic region were found in 62.7, 21.9, and 30% of 169 genotypically distinct isoniazid-resistant isolates, respectively, whereas they were found in 0, 0, and 8% of susceptible strains, respectively. The frequency of mutation at each locus was unrelated to the resistance profile or previous antituberculous drug therapy. The commonest mutation in the oxyR-ahpC intergenic region, ahpC -46A, was present in 23.7% of isoniazid-resistant isolates and 7.5% of susceptible isolates. This proved to be a phylogenetic marker for a subgroup of M. tuberculosis strains originating on the Indian subcontinent, which shared IS6110-based restriction fragment length polymorphism and spoligotype features with the Delhi strain and Central Asian strain CAS1; and this marker is strongly associated with isoniazid resistance and the katG 315Thr mutation. In total, 82.8% of unrelated isoniazid-resistant isolates could be identified by analysis of just two loci: katG 315 and the inhA promoter. Analysis of the oxyR-ahpC intergenic region, although phylogenetically interesting, does not contribute significantly to further identification of isoniazid-resistant isolates. PMID- 15793127 TI - Effect of mupirocin treatment on nasal, pharyngeal, and perineal carriage of Staphylococcus aureus in healthy adults. AB - Nasal carriage of Staphylococcus aureus is an important risk factor for S. aureus infections. Mupirocin nasal ointment is presently the treatment of choice for decolonizing the anterior nares. However, recent clinical trials show limited benefit from mupirocin prophylaxis in preventing nosocomial S. aureus infections, probably due to (re)colonization from extranasal carriage sites. Therefore, we studied the effectiveness of mupirocin nasal ointment treatment on the dynamics of S. aureus nasal and extranasal carriage. Twenty noncarriers, 26 intermittent carriers, and 16 persistent carriers had nasal, throat, and perineum samples taken 1 day before and 5 weeks after mupirocin treatment (twice daily for 5 days) and assessed for growth of S. aureus. The identities of cultured strains were assessed by restriction fragment length polymorphisms of the coagulase and protein A genes. The overall carriage rate (either nasal, pharyngeal, or perineal carrier or a combination) was significantly reduced after mupirocin treatment from 30 to 17 carriers (P = 0.003). Of the 17 carriers, 10 (60%) were still colonized with their old strain, 6 (35%) were colonized with an exogenous strain, and 1 (5%) was colonized with both. Two noncarriers became carriers after treatment. The acquisition of exogenous strains after mupirocin treatment is a common phenomenon. The finding warrants the use of mupirocin only in proven carriers for decolonization purposes. Mupirocin is effective overall in decolonizing nasal carriers but less effective in decolonizing extranasal sites. PMID- 15793128 TI - Comparative antimicrobial characterization of LBM415 (NVP PDF-713), a new peptide deformylase inhibitor of clinical importance. AB - LBM415 (NVP PDF-713) is the first member of the peptide deformylase (PDF) inhibitor class being developed for clinical trials as a parenteral and oral agent for treatment of community-acquired respiratory tract disease and serious infections caused by antimicrobial-resistant gram-positive cocci. In this study susceptibility testing results from 1,306 recent clinical isolates selected to over-represent resistance trends among the species were summarized. All staphylococci (153 strains; MIC at which 90% of isolates were inhibited [MIC90], 2 microg/ml), Streptococcus pneumoniae (170 strains; MIC90, 1 microg/ml), other streptococci (150 strains; MIC90, 1 microg/ml), enterococci (104 strains; MIC90, 4 microg/ml), Moraxella catarrhalis (103 strains; MIC90, 0.5 microg/ml), and Legionella pneumophila (50 strains; MIC90, 0.12 microg/ml) were inhibited at < or = 8 microg of LBM415/ml, as were 97% of Haemophilus influenzae isolates (300 strains; MIC90, 4 to 8 microg/ml). Among other bacterial groups, 100% of gram positive and -negative anaerobes, including 22 Bacteroides spp. strains (31 strains total; MIC90, 1 microg/ml), were inhibited by < or = 4 microg/ml, whereas Enterobacteriaceae (112 strains) and most nonfermentative bacilli (107 strains) were not inhibited at readily achievable concentrations. The compound was found to have a dominantly bacteriostatic action, and spontaneous single-step mutational rates occurred at low levels (10(-6) to <10(-8)). Drug interaction studies failed to identify any class-specific synergistic interactions, nor were antagonistic interactions observed. Variations in broth and agar MIC test conditions demonstrated that, whereas the agar-based method trended towards a 1 log2 dilution-higher MIC than the broth method and was inoculum dependent, other variations in incubation environment, medium supplements, pH, or calcium concentration had little influence on LBM415 MIC results. Use of the efflux inhibitor phe-arg-beta-naphthylamide showed an average of 1 log2 dilution decrease in H. influenzae MICs, demonstrating the contribution of efflux pumps in influencing susceptibility to PDF inhibitors. The in vitro activity of LBM415 against targeted bacterial species, including resistant subsets, and other laboratory characteristics of this novel compound demonstrate the potential of PDF inhibitors as a new class of antimicrobial agents. PMID- 15793129 TI - A Helicobacter pylori TolC efflux pump confers resistance to metronidazole. AB - In Helicobacter pylori, the contribution of efflux proteins to antibiotic resistance is not well established. As translocases that act in parallel may have overlapping substrate specificities, the loss of function of one such translocase may be compensated for by that of another translocase with no effect on susceptibilities to antibiotics. The genome of H. pylori 26695 was assessed for the presence of putative translocases and outer membrane efflux or TolC-like proteins which could interact to form efflux systems involved in drug resistance. Twenty-seven translocases were identified, of which HP1184 was the sole representative of the multidrug and toxic compound extrusion family of translocases and which could thus have a unique substrate specificity. In addition, four TolC-like proteins (HP0605, HP0971, HP1327, and HP1489) were identified. Thus, it is feasible that inactivation of a TolC-like protein would affect the functions of multiple translocases. We aimed to determine whether efflux systems contribute to antimicrobial susceptibility by evaluation of the susceptibility profiles of an HP1184-knockout mutant, four mutants in which one of the four TolC homologs was inactivated, as well as a mutant in which both HP0605 and HP0971 were inactivated. The HP1184- and HP1489-knockout mutants both showed increased susceptibilities to ethidium bromide, while the HP0605-knockout mutant exhibited increased susceptibilities to novobiocin and sodium deoxycholate. The HP0605 and HP0971 double-knockout mutant was also more susceptible to metronidazole, in addition to being susceptible to novobiocin and sodium deoxycholate. Thus, active efflux is an eminent means of resistance to antimicrobials in H. pylori and resembles the situation in other bacteria. PMID- 15793130 TI - Phenotypic tolerance: antibiotic enrichment of noninherited resistance in bacterial populations. AB - When growing bacteria are exposed to bactericidal concentrations of antibiotics, the sensitivity of the bacteria to the antibiotic commonly decreases with time, and substantial fractions of the bacteria survive. Using Escherichia coli CAB1 and antibiotics of five different classes (ampicillin, ciprofloxacin, rifampin, streptomycin, and tetracycline), we examine the details of this phenomenon and, with the aid of mathematical models, develop and explore the properties and predictions of three hypotheses that can account for this phenomenon: (i) antibiotic decay, (ii) inherited resistance, and (iii) phenotypic tolerance. Our experiments cause us to reject the first two hypotheses and provide evidence that this phenomenon can be accounted for by the antibiotic-mediated enrichment of subpopulations physiologically tolerant to but genetically susceptible to these antibiotics, phenotypic tolerance. We demonstrate that tolerant subpopulations generated by exposure to one concentration of an antibiotic are also tolerant to higher concentrations of the same antibiotic and can be tolerant to antibiotics of the other four types. Using a mathematical model, we explore the effects of phenotypic tolerance to the microbiological outcome of antibiotic treatment and demonstrate, a priori, that it can have a profound effect on the rate of clearance of the bacteria and under some conditions can prevent clearance that would be achieved in the absence of tolerance. PMID- 15793131 TI - Cloning, sequencing, and characterization of the SdeAB multidrug efflux pump of Serratia marcescens. AB - Serratia marcescens is an important nosocomial agent known for causing various infections in immunocompromised individuals. Resistance of this organism to a broad spectrum of antibiotics makes the treatment of infections very difficult. This study was undertaken to identify multidrug resistance efflux pumps in S. marcescens. Three mutant strains of S. marcescens were isolated in vitro by the serial passaging of a wild-type strain in culture medium supplemented with ciprofloxacin, norfloxacin, or ofloxacin. Fluoroquinolone accumulation assays were performed to detect the presence of a proton gradient-dependent efflux mechanism. Two of the mutant strains were found to be effluxing norfloxacin, ciprofloxacin, and ofloxacin, while the third was found to efflux only ofloxacin. A genomic library of S. marcescens wild-type strain UOC-67 was constructed and screened for RND pump-encoding genes by using DNA probes for two putative RND pump-encoding genes. Two different loci were identified: sdeAB, encoding an MFP and an RND pump, and sdeCDE, encoding an MFP and two different RND pumps. Northern blot analysis revealed overexpression of sdeB in two mutant strains effluxing fluoroquinolones. Analysis of the sdeAB and sdeCDE loci in Escherichia coli strain AG102MB, deficient in the RND pump (AcrB), revealed that gene products of sdeAB are responsible for the efflux of a diverse range of substrates that includes ciprofloxacin, norfloxacin, ofloxacin, chloramphenicol, sodium dodecyl sulfate, ethidium bromide, and n-hexane, while those of sdeCDE did not result in any change in susceptibilities to any of these agents. PMID- 15793132 TI - Efficacy of ciprofloxacin-releasing bioabsorbable osteoconductive bone defect filler for treatment of experimental osteomyelitis due to Staphylococcus aureus. AB - The concept of local antibiotic delivery via biodegradable bone defect fillers with multifunctional properties for the treatment of bone infections is highly appealing. Fillers can be used to obliterate surgical dead space and to provide targeted local bactericidal concentrations in tissue for extended periods. Eventually, the osteoconductive component of the filler could guide the healing of the bone defect. The present experimental study was carried out to test this concept in a localized Staphylococcus aureus osteomyelitis model in the rabbit (n = 31). A metaphyseal defect of the tibia was filled with a block of bone cement, followed by insertion of a bacterial inoculum. After removal of the bone cement and surgical debridement at 2 weeks, the defect was filled with a ciprofloxacin containing (7.6% +/- 0.1%, by weight) composite (treated-infection group) or with a composite without antibiotic (sham-treated group). Both a positive control group (untreated-infection group) and a negative control group were also produced. The treatment response, monitored by positron emission tomography (PET) with fluorine-18-labeled fluorodeoxyglucose ([18F]FDG) at 3 and 6 weeks, showed rapidly decreasing amounts of [18F]FDG uptake in the treated-infection group (P = 0.001 compared with the results for the untreated-infection group at 6 weeks). The bacteriological analysis confirmed the eradication of the bone pathogen in the treated-infection group. However, three animals had culture-positive soft tissue infections. All animals in the sham-treated and untreated-infection groups had culture-positive bone infections with typical radiographic changes of osteomyelitis. Histomorphometry, peripheral quantitative computed tomography, and backscattered electron imaging of scanning electron microscopy images verified the osteoconductive properties of the bioactive glass microspheres within the composite. The median bone ciprofloxacin concentrations were 1.2 and 2.1 microg/g at two anatomic locations of the tibia. This is the first report to show the value of [18F]FDG PET for quantitative monitoring of the treatment response in bone infections. The collaborative results of bacteriologic and [18F-FDG] PET studies showed that use of the multifunctional composite was successful for eradication of the S. aureus pathogen from bone. PMID- 15793133 TI - Comparative safety evaluation of the candidate vaginal microbicide C31G. AB - C31G is currently the focus of clinical trials designed to evaluate this agent as a microbicidal and spermicidal agent. In the following studies, the in vivo safety of C31G was assessed with a Swiss Webster mouse model of cervicovaginal toxicity and correlated with results from in vitro cytotoxicity experiments and published clinical observations. A single exposure of unformulated 1% C31G resulted in mild-to-moderate epithelial disruption and inflammation at 2 and 4 h postapplication. The columnar epithelium of the cervix was the primary site of damage, while no perturbation of the vaginal mucosa was observed. In contrast, application of unformulated 1.7% C31G resulted in greater levels of inflammation in the cervical epithelium at 2 h postapplication and severe epithelial disruption that persisted to 8 h postapplication. Application of a nonionic aqueous gel formulation containing 1% C31G resulted in no apparent cervicovaginal toxicity at any time point evaluated. However, formulation of 1.7% C31G did not substantially reduce the toxicity associated with unformulated C31G at that concentration. These observations correlate with findings gathered during a recent clinical trial, in which once-daily applications resulted in no adverse events in women receiving the formulation containing 1% C31G, compared to moderate-to-severe adverse events in 30% of women receiving the 1.7% C31G formulation. The Swiss Webster mouse model was able to effectively discriminate between concentrations and formulations of C31G that produced distinct clinical effects in human trials. The Swiss Webster animal model may be a highly valuable tool for preclinical evaluation of candidate vaginal microbicides. PMID- 15793134 TI - Treatment with benznidazole during the chronic phase of experimental Chagas' disease decreases cardiac alterations. AB - Chagas' disease, caused by Trypanosoma cruzi infection, is one of the main causes of death due to heart failure in Latin American countries. Benznidazole, the chemotherapeutic agent most often used for the treatment of chagasic patients, is highly toxic and has limited efficacy, especially in the chronic phase of the disease. In the present study we used a mouse model of chronic Chagas' disease to investigate the effects of benznidazole treatment during the chronic phase on disease progression. The hearts of benznidazole-treated mice had decreased parasitism and myocarditis compared to the hearts of untreated chagasic mice. Both groups of Trypanosoma cruzi-infected mice had significant alterations in their electrocardiograms compared to those of the healthy mice. However, untreated mice had significantly higher cardiac conduction disturbances than benznidazole-treated mice, including intraventricular conduction disturbances, atrioventricular blocks, and extrasystoles. The levels of antibodies against T. cruzi antigens (epimastigote extract, P2beta, and trans-sialidase) as well as antibodies against peptides of the second extracellular loops of beta1-adrenergic and M2-muscarinic cardiac receptors were also lower in the sera from benznidazole treated mice than in the sera from untreated mice. These results demonstrate that treatment with benznidazole in the chronic phase of infection prevents the development of severe chronic cardiomyopathy, despite the lack of complete parasite eradication. In addition, our data highlight the role of parasite persistence in the development of chronic Chagas' disease and reinforce the importance of T. cruzi elimination in order to decrease or prevent the development of severe chagasic cardiomyopathy. PMID- 15793135 TI - Expression of ccaR, encoding the positive activator of cephamycin C and clavulanic acid production in Streptomyces clavuligerus, is dependent on bldG. AB - In Streptomyces coelicolor, bldG encodes a putative anti-anti-sigma factor that regulates both aerial hypha formation and antibiotic production, and a downstream transcriptionally linked open reading frame (orf3) encodes a putative anti-sigma factor protein. A cloned DNA fragment from Streptomyces clavuligerus contained an open reading frame that encoded a protein showing 92% identity to the S. coelicolor BldG protein and 91% identity to the BldG ortholog in Streptomyces avermitilis. Sequencing of the region downstream of bldG in S. clavuligerus revealed the presence of an open reading frame encoding a protein showing 72 and 69% identity to the ORF3 proteins in S. coelicolor and S. avermitilis, respectively. Northern analysis indicated that, as in S. coelicolor, the S. clavuligerus bldG gene is expressed as both a monocistronic and a polycistronic transcript, the latter including the downstream orf3 gene. High-resolution S1 nuclease mapping of S. clavuligerus bldG transcripts revealed the presence of three bldG-specific promoters, and analysis of expression of a bldGp-egfp reporter indicated that the bldG promoter is active at various stages of development and in both substrate and aerial hyphae. A bldG null mutant was defective in both morphological differentiation and in the production of secondary metabolites, such as cephamycin C, clavulanic acid, and the 5S clavams. This inability to produce cephamycin C and clavulanic acid was due to the absence of the CcaR transcriptional regulator, which controls the expression of biosynthetic genes for both secondary metabolites as well as the expression of a second regulator of clavulanic acid biosynthesis, ClaR. This makes bldG the first regulatory protein identified in S. clavuligerus that functions upstream of CcaR and ClaR in a regulatory cascade to control secondary metabolite production. PMID- 15793137 TI - Molecular mechanisms by which rRNA mutations confer resistance to clindamycin. AB - The mechanisms by which rRNA mutations confer clindamycin resistance were examined in Mycobacterium smegmatis strains containing homogeneous populations of ribosomes with base substitutions at nucleotides A2058 and A2059. Computer graphic predictions based on structural studies correlate with the resistance phenotypes for six of seven strains with unique rRNA mutations. PMID- 15793136 TI - Photodynamic effects of novel XF porphyrin derivatives on prokaryotic and eukaryotic cells. AB - The worldwide rise in the rates of antibiotic resistance of bacteria underlines the need for alternative antibacterial agents. A promising approach to the killing of gram-positive antibiotic-resistant bacteria of the skin uses light in combination with a photosensitizer to induce a phototoxic reaction. Different concentrations (0 to 100 microM) of porphyrin-based photosensitizers (CTP1, XF70, and XF73) and different incubation times (5 min, 1 h, and 4 h) were used to determine phototoxicity against two methicillin-resistant Staphylococcus aureus strains, one methicillin-sensitive S. aureus strain, one methicillin-resistant Staphylococcus epidermidis strain, one Escherichia coli strain, and human keratinocytes and fibroblasts. Incubation with 0.005 microM XF70 or XF73, followed by illumination, yielded a 3-log10 (> or = 99.9%) decrease in the viable cell numbers of all staphylococcal strains, indicating that the XF drugs have high degrees of potency against gram-positive bacteria and also that the activities of these novel drugs are independent of the antibiotic resistance pattern of the staphylococci examined. CTP1 was less potent against the staphylococci under the same conditions. At 0.005 microM, XF70 and XF73 demonstrated no toxicity toward fibroblasts or keratinocytes. No inactivation of E. coli was detected at this concentration. XF73 was confirmed to act via a reactive oxygen species from the results of studies with sodium azide (a quencher of singlet oxygen), which reduced the killing of both eukaryotic and prokaryotic cells. When a quencher of superoxide anion and the hydroxyl radical was used, cell killing was not inhibited. These results demonstrate that the porphyrin based photosensitizers had concentration-dependent differences in their efficacies of killing of methicillin-resistant staphylococcal strains via reactive oxygen species without harming eukaryotic cells at the same concentrations. PMID- 15793138 TI - Comparative efficacies of TAK-187, a long-lasting ergosterol biosynthesis inhibitor, and benznidazole in preventing cardiac damage in a murine model of Chagas' disease. AB - We carried out a comparative study of benznidazole and TAK-187, a long-lasting ergosterol biosynthesis inhibitor, with a murine model of Chagas' disease. The results indicated that TAK-187 was more effective than benznidazole in preventing Trypanosoma cruzi-induced cardiac damage in experimental animals. PMID- 15793139 TI - Penicillin-binding proteins in the pathogenic intestinal spirochete Brachyspira pilosicoli. AB - Penicillin-binding proteins (PBPs) of slightly different molecular masses (94, 62 or 68, 42 or 50, 25, and 22 kDa) were identified in one human and two porcine Brachyspira pilosicoli strains. Identification of PBPs of B. pilosicoli provides a basis for characterization of the genes encoding these proteins among pathogenic intestinal spirochetes of humans and animals. PMID- 15793140 TI - Novel TEM-type extended-spectrum beta-lactamase, TEM-134, in a Citrobacter koseri clinical isolate. AB - A new natural TEM derivative with extended-spectrum beta-lactamase activity, TEM 134, was identified in a ceftazidime-resistant clinical isolate of Citrobacter koseri. Compared to TEM-1, TEM-134 contains the following mutations: Q39K, E104K, R164H, and G238S. The bla(TEM-134) gene was not transferable by conjugation and, apparently, was chromosomally encoded. Expression studies with Escherichia coli revealed efficient cefotaximase and ceftazidimase activity for TEM-134. PMID- 15793141 TI - CTX-M-10 linked to a phage-related element is widely disseminated among Enterobacteriaceae in a Spanish hospital. AB - CTX-M-10 has been widely disseminated among multiple clones of several species of Enterobacteriaceae, harboring seemingly different plasmids, for over a decade in Ramon y Cajal University Hospital, Madrid, Spain. Cloning and sequencing of a 12.2-kb DNA fragment from plasmid pRYCE21 from Klebsiella pneumoniae strain KP4aC revealed a novel phage-related element immediately upstream of bla(CTX-M-10) conserved among different CTX-M-10-producing strains. This is the first report showing an extended-spectrum-beta-lactamase gene linked to a phage-related element. PMID- 15793142 TI - Occurrence of CTX-M-3, CTX-M-15, CTX-M-14, and CTX-M-9 extended-spectrum beta lactamases in Enterobacteriaceae clinical isolates in Korea. AB - Among 603 isolates of Enterobacteriaceae collected between June and November 2003 from three university hospitals within Korea, bla(CTX-M-3), bla(CTX-M-15), bla(CTX-M-14), and bla(CTX-M-9) were detected in 41 isolates of species from five different genera of Enterobacteriaceae, Escherichia coli, Klebsiella pneumoniae, Citrobacter freundii, Enterobacter spp., and Serratia marcescens. PMID- 15793143 TI - Excision of nucleoside analogs from DNA by p53 protein, a potential cellular mechanism of resistance to inhibitors of human immunodeficiency virus type 1 reverse transcriptase. AB - We investigated the ability of p53 in cytoplasm to excise nucleoside analogs (NAs). A decrease in incorporation of NAs by human immunodeficiency virus type 1 reverse transcriptase and their excision from DNA by p53, provided by the cytoplasmic fraction of LCC2 cells, suggest that p53 in cytoplasm may act as an external proofreader for NA incorporation. PMID- 15793145 TI - In vitro effects of spectinomycin and ceftriaxone alone or in combination with other antibiotics against Chlamydia trachomatis. AB - The in vitro effects of spectinomycin and ceftriaxone, alone or in combination with erythromycin, ofloxacin, and doxycycline, against Chlamydia trachomatis were investigated by the checkerboard method and compared by Ridit (reference identical unit) analysis. A combination of spectinomycin with erythromycin or doxycycline was found to be more effective than that of ceftriaxone. PMID- 15793144 TI - Early selection of a new UL97 mutant with a severe defect of ganciclovir phosphorylation after valaciclovir prophylaxis and short-term ganciclovir therapy in a renal transplant recipient. AB - We describe the emergence of a new ganciclovir resistance mutation in the UL97 gene of human cytomegalovirus, deletion of codon 601, after valaciclovir and short-term ganciclovir therapy following kidney transplantation. Its role in ganciclovir resistance was supported by decreased ganciclovir phosphorylation in a recombinant vaccinia virus system. PMID- 15793146 TI - Different rifampin sensitivities of Escherichia coli and Mycobacterium tuberculosis RNA polymerases are not explained by the difference in the beta subunit rifampin regions I and II. AB - Mycobacterium tuberculosis RNA polymerase is 1,000-fold more sensitive to rifampin than Escherichia coli RNA polymerase. Chimeric E. coli RNA polymerase in which the beta-subunit segment encompassing rifampin regions I and II (amino acids [aa] 463 through 590) was replaced with the corresponding region from M. tuberculosis (aa 382 through 509) did not show an increased sensitivity to the antibiotic. Thus, the difference in amino acid sequence between the rifampin regions I and II of the two species does not account for the difference in rifampin sensitivity of the two polymerases. PMID- 15793147 TI - Capsule homology does not increase the frequency of transformation of linked penicillin binding proteins PBP 1a and PBP 2x in Streptococcus pneumoniae. AB - Penicillin resistance is mainly confined to a limited number of Streptococcus pneumoniae serotypes. Given linkage between the capsular biosynthesis locus and two penicillin binding proteins, we tested whether capsule homology increases transformation rates of penicillin resistance. Transformation rates in homologous donor-recipient pairs were no higher than expected, falsifying this hypothesis. PMID- 15793148 TI - Synergistic activities of fluconazole and voriconazole with terbinafine against four Candida species determined by checkerboard, time-kill, and Etest methods. AB - The in vitro activities of fluconazole or voriconazole plus terbinafine were evaluated against 20 Candida isolates by the checkerboard, time-kill, and Etest methods. Synergism (C. albicans, C. glabrata, and C. tropicalis) and indifference (C. krusei) were observed. Correlation among methods was good. The Etest is a suitable method to determine drug interactions. PMID- 15793149 TI - Antifungal activity of flocculosin, a novel glycolipid isolated from Pseudozyma flocculosa. AB - Flocculosin, a glycolipid isolated from the yeast-like fungus Pseudozyma flocculosa, was investigated for in vitro antifungal activity. The compound displayed antifungal properties against several pathogenic yeasts. Synergistic activity was observed between flocculosin and amphotericin B, and no significant cytotoxicity was demonstrated when tested against human cell lines. PMID- 15793150 TI - Impact of clarithromycin resistance on eradication of Helicobacter pylori in infected adults. AB - The outcome of Helicobacter pylori infection was analyzed in 114 dyspeptic patients treated with triple-drug therapy including clarithromycin. Clarithromycin resistance (in 20.2% of our isolates) was mainly caused by an A2142G mutation in the 23S rRNA gene of H. pylori. H. pylori eradication was obtained in all patients with clarithromycin-susceptible isolates but not in any patients with clarithromycin-resistant isolates (P = 0.0001). Therefore, it would be useful to conduct H. pylori antimicrobial susceptibility testing of the first gastric biopsy culture before choosing the first three drugs for therapy of infected patients. PMID- 15793151 TI - Sensititre YeastOne caspofungin susceptibility testing of Candida clinical isolates: correlation with results of NCCLS M27-A2 multicenter study. AB - The ability of Sensititre YeastOne to discriminate isolates with reduced caspofungin susceptibility was determined against 36 Candida spp. (6 with a known FKS1 mutation). Results were compared with those of M27-A2. The MIC endpoint was 100% growth inhibition. Overall agreement (+/-2 log2) was 87.16%. Sensititre YeastOne detected strains with reduced caspofungin susceptibility. PMID- 15793153 TI - Antibiotic tolerance induced by lactoferrin in clinical Pseudomonas aeruginosa isolates from cystic fibrosis patients. AB - Lactoferrin-induced cell depolarization and a delayed tobramycin-killing effect on Pseudomonas aeruginosa cells were correlated. This antibiotic tolerance effect (ATE) reflects the ability of a defense protein to modify the activity of an antibiotic as a result of its modulatory effect on bacterial physiology. P. aeruginosa isolates from cystic fibrosis patients showed higher ATE values (< or = 6-fold) than other clinical strains. PMID- 15793152 TI - Cloning, characterization, and expression analysis of hepcidin gene from red sea bream (Chrysophrys major). AB - A cDNA encoding hepcidin was isolated from a library of cDNA from spleen of red sea bream (Chrysophrys major) by expressed sequence tag analysis. The expression of the hepcidin mRNA in various tissues was examined. Challenge of red sea bream with Escherichia coli DH5alpha elevated hepcidin mRNA levels in spleen, gill, liver, and intestine. PMID- 15793154 TI - Secretion of proinflammatory cytokines and chemokines during amphotericin B exposure is mediated by coactivation of toll-like receptors 1 and 2. AB - Amphotericin B (AmB) is a ligand of toll-like receptor 2 (TLR2). Here, we demonstrate the participation of TLR1 in AmB-induced cell activation that led to the secretion of tumor necrosis factor alpha, interleukin 6 (IL-6), and IL-8. Hence, TLR2-TLR1 coactivation serves as the underlying mechanism for the proinflammatory toxicities associated with AmB. PMID- 15793155 TI - Pharmacokinetics of oral doxycycline during combination treatment of severe falciparum malaria. AB - The pharmacokinetics of oral doxycycline administered at 200 mg every 24 h were investigated in 17 patients recovering from severe Plasmodium falciparum malaria. The data suggest that the doses of doxycycline currently recommended (circa 3.5 mg/kg of body weight daily) may not be optimal. PMID- 15793156 TI - Role of P-glycoprotein in distribution of nelfinavir across the blood-mammary tissue barrier and blood-brain barrier. AB - As a first approach in understanding the possible efficacy and toxicity of human immunodeficiency virus protease inhibitors during breast feeding, the milk-to plasma ratio of nelfinavir was determined in lactating rats. The milk-to-plasma ratio of nelfinavir was determined to be 0.56 +/- 0.10 (means +/- standard deviations). Western blotting indicated that P-glycoprotein is expressed in rat mammary and brain tissue; however, the multidrug-resistant modulator GF120918 showed a significant effect only at the blood-brain barrier and not at the mammary-epithelial tissue barrier. PMID- 15793157 TI - Pharmacokinetic profile of tigecycline in serum and skin blister fluid of healthy subjects after multiple intravenous administrations. AB - The pharmacokinetics of tigecycline, when given as a 100-mg loading dose followed by 50 mg every 12 h, were determined in serum and blister fluid. The peak tigecycline concentration and half-life in serum were greater than those in blister fluid. Tigecycline penetrates into blister fluid well, with a mean penetration rate of 74%. PMID- 15793159 TI - Presence of tetracycline resistance determinants and susceptibility to tigecycline and minocycline. AB - No relation between the presence of tetracycline resistance determinants tet(A) to tet(E) and the MICs of tigecycline was observed for Enterobacteriaceae, although tetracycline-susceptible isolates were more susceptible overall to tigecycline, whereas the presence of tet(M) in Staphylococcus aureus was associated with higher MICs of minocycline. PMID- 15793158 TI - Mutant prevention concentrations of ABT-492, levofloxacin, moxifloxacin, and gatifloxacin against three common respiratory pathogens. AB - The purpose of this study was to compare the mutant prevention concentration (MPC) of ABT-492 to those of levofloxacin, moxifloxacin, and gatifloxacin against Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. The fluoroquinolones had comparable mutation selection windows, which is the ratio of MPC/MIC, for all isolates. PMID- 15793160 TI - Functional characterization of OXA-57, a class D beta-lactamase from Burkholderia pseudomallei. AB - Class D beta-lactamase OXA-57 was identified in a range of isolates of Burkholderia pseudomallei and Burkholderia thailandensis. Comparative kinetic analyses of wild-type and mutant forms of B. pseudomallei OXA-57 are reported. Implications of these data for beta-lactam resistance and the proposed role of Ser-104 in beta-lactam hydrolysis are discussed. PMID- 15793161 TI - Efficacy and safety of generic amphotericin B in experimental pulmonary aspergillosis. AB - The recent shortage of the brand name drug Fungizone has necessitated a change to generic formulations of amphotericin B deoxycholate. Clinical trials cannot be conducted in a timely manner to provide data on the safety and efficacy of these formulations. We therefore compared generic amphotericin B and Fungizone for activity and safety in the treatment of experimental invasive pulmonary aspergillosis (IPA) in persistently neutropenic rabbits. Fungizone and generic amphotericin B are similar in efficacy, pharmacokinetics, and safety in the treatment of experimental IPA. PMID- 15793162 TI - Genetic basis of antibiotic resistance in clinical isolates of Streptococcus gallolyticus (Streptococcus bovis). AB - Among 128 Streptococcus gallolyticus (Streptococcus bovis) isolates, 77.7% were resistant to tetracyclines and contained tet(M) and/or tet(L) and/or tet(O). A total of 59.4% had macrolide resistance and contained erm(B) and, rarely, mef(A). Among the one-third of isolates highly resistant to kanamycin and streptomycin, most harbored aphA3 and aad-6 genes. PMID- 15793163 TI - Levofloxacin and ciprofloxacin decrease procainamide and N-acetylprocainamide renal clearances. AB - Ten healthy adults participated in a randomized, crossover drug interaction study testing procainamide only, procainamide plus levofloxacin, and procainamide plus ciprofloxacin. During levofloxacin therapy, most procainamide and N acetylprocainamide (NAPA) pharmacokinetic parameters, including decreased renal clearances and renal clearance/creatinine clearance ratios, changed (P < 0.05). During ciprofloxacin treatment, only procainamide and NAPA renal clearances decreased significantly. PMID- 15793164 TI - In vitro and in vivo cytotoxicities and antileishmanial activities of thymol and hemisynthetic derivatives. AB - The in vitro and in vivo antileishmanial and cytotoxic activities of thymol and structural derivatives in comparison to those of Glucantime were studied. The results showed here suggest that thymol and hemisynthetic derivatives have promising antileishmanial potential and could be considered as new lead structures in the search for novel antileishmanial drugs. PMID- 15793165 TI - Effects of age and sex on single-dose pharmacokinetics of tigecycline in healthy subjects. AB - The pharmacokinetics of tigecycline was evaluated in 46 healthy young and elderly men and women. Except for the volumes of distribution at steady state (approximately 350 liters in women versus 500 liters in men), there were no significant differences in tigecycline pharmacokinetic parameters. Based on pharmacokinetics, no dosage adjustment is warranted based on age or sex. PMID- 15793166 TI - Alteration in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV in Quinolone-resistant Shigella dysenteriae serotype 1 clinical isolates from Kolkata, India. PMID- 15793167 TI - Rollback of Salmonella enterica serotype Typhi resistance to chloramphenicol and other antimicrobials in Kolkata, India. PMID- 15793169 TI - Nucleotide sequence of IS1678, an insertion sequence in the vanA cluster of enterococci. PMID- 15793168 TI - Emergence of daptomycin resistance in Enterococcus faecium during daptomycin therapy. PMID- 15793170 TI - Detection of VIM-2 metallo-beta-lactamase in Pseudomonas aeruginosa from Germany. PMID- 15793171 TI - Effect of a pharmaceutical care program on vascular risk factors in type 2 diabetes: the Fremantle Diabetes Study. AB - OBJECTIVE: To examine the effect of a 12-month pharmaceutical care (PC) program on vascular risk in type 2 diabetes. RESEARCH DESIGN AND METHODS: We recruited 198 community-based patients randomized to PC or usual care. PC patients had face to-face goal-directed medication and lifestyle counseling at baseline and at 6 and 12 months plus 6-weekly telephone assessments and provision of other educational material. Clinical, biochemical, and medication-related data were sent regularly to each patient's physician(s). The main outcome measure was change in HbA(1c). A diabetes-specific risk engine was used to estimate changes in 10-year coronary heart disease (CHD) and stroke risk in patients without a history of cardiovascular disease. RESULTS: At total of 180 patients (91%) completed the study. Mean (95% CI) reductions were greater in PC case subjects (n = 92) than control subjects (n = 88) for HbA(1c) (-0.5% [95% CI -0.7 to -0.3] vs. 0 [-0.2 to 0.2]) and systolic (-14 mmHg [-19 to -9] vs. -7 [-11 to -2]) and diastolic (-5 mmHg [-8 to -3] vs. -2 [-4 to 1]) blood pressure (P < or = 0.043). The improvement in HbA(1c) persisted after adjustment for baseline value and demographic and treatment-specific variables. The median (interquartile range) 10 year estimated risk of a first CHD event decreased in the PC case subjects (25.1% [15.6-36.2] to 20.3 [14.6-30.2]; n = 42, P = 0.002) but not in the control subjects (26.1% [17.2-39.4] vs. 26.4 [16.7-38.0]; n = 52, P = 0.17). CONCLUSIONS: A 12-month PC program in type 2 diabetes reduced glycemia and blood pressure. Pharmacist involvement contributed to improvement in HbA(1c) independently of pharmacotherapeutic changes. PC could prove a valuable component of community based multidisciplinary diabetes care. PMID- 15793173 TI - Generalizability and persistence of a multifaceted intervention for improving quality of care for rural patients with type 2 diabetes. AB - OBJECTIVE: Most quality improvement efforts for type 2 diabetes have neglected cardiovascular risk factors and are limited by a lack of information about generalizability across settings or persistence of effect over time. RESEARCH DESIGN AND METHODS: We previously reported 6-month results of a controlled study of an intervention that improved cardiovascular risk factors for rural patients with type 2 diabetes. We subsequently provided the identical intervention to the control region after the main study was completed. The primary outcome was 10% improvement in systolic blood pressure, total cholesterol, or HbA(1c). We compared the previously reported 6-month effect of the original intervention with the effect of the crossed-over intervention to the former control region and remeasured outcomes in the original intervention region 12 months later. RESULTS: Our analysis included 200 original intervention and 181 crossed-over intervention subjects. The age of the population was 62.4 +/- 12.4 years (mean +/- SD), and 54.3% were women. A similar proportion of patients in the crossed-over intervention group achieved improvement in the primary composite outcome compared with the original intervention group (38 vs. 44%, respectively; P = 0.29). In adjusted analyses, we observed less improvement in blood pressure (adjusted odds ratio 0.40 [95% CI 0.17-0.75]) but greater improvements in total cholesterol (1.86 [0.93-3.7]) with the crossed-over intervention compared with the original intervention. We observed sustained improvements in total cholesterol and HbA(1c) levels in the original intervention group, whereas previous large gains in control of blood pressure diminished over time. CONCLUSIONS: We found that our intervention was generalizable across settings, and its effect persisted over time. Nevertheless, without ongoing intervention or reinforcement, we noted some loss of the original benefits that had accrued. Future translational work should incorporate interventions such as ours into ongoing systems of rural care. PMID- 15793172 TI - Leisure time physical activity is associated with poor glycemic control in type 1 diabetic women: the FinnDiane study. AB - OBJECTIVE: We studied the association between leisure time physical activity (LTPA) and glycemic control, insulin dose, and estimated glucose disposal rate (eGDR) in type 1 diabetes. RESEARCH DESIGN AND METHODS: This is a cross-sectional study of 1,030 type 1 diabetic patients participating in the Finnish Diabetic Nephropathy Study, a nationwide multicenter study. LTPA was assessed by a validated 12-month questionnaire and expressed in metabolic equivalent (MET) units. Patients were grouped as sedentary (LTPA <10 MET h/week, n = 247), moderately active (LTPA 10-40 MET h/week, n = 568), and active (LTPA >40 MET h/week, n = 215). Outcome measures were HbA(1c), insulin dose, and eGDR (estimate of insulin sensitivity based on waist-to-hip ratio, hypertension, and HbA(1c)). RESULTS: LTPA correlated with HbA(1c) in women (r = -0.12, P = 0.007) but not in men (r = -0.03, P = 0.592). Sedentary women had higher HbA(1c) than moderately active and active women: 8.8 +/- 1.4% vs. 8.3 +/- 1.4% vs. 8.3 +/- 1.4% (P = 0.004), whereas HbA(1c) in men was 8.4 +/- 1.3% vs. 8.2 +/- 1.4% vs. 8.2 +/- 1.3% (P = 0.774), respectively. In men, insulin doses were 0.74 +/- 0.21 vs. 0.71 +/- 0.20 vs. 0.68 +/- 0.23 IU . kg(-1) . 24 h(-1) (P = 0.003). In both sexes, sedentary patients had lower eGDRs than active patients [median (interquartile range) 5.5 (4.0-8.2) vs. 6.8 (4.7-8.8) vs. 6.7 (4.6-8.6) mg . kg(-1) . min(-1); P < 0.01 for sedentary vs. others]. Age, obesity, smoking, insulin dose, social class, diabetic nephropathy, or cardiovascular disease did not explain the results. CONCLUSIONS: Low levels of LTPA were associated with poor glycemic control in type 1 diabetic women. Men seem to use less insulin when physically active. Increased LTPA levels were associated with increased estimated insulin sensitivity. Longitudinal studies are needed to further clarify the effects of LTPA on type 1 diabetes. PMID- 15793174 TI - Impact of fasting plasma glucose levels on gastric cancer incidence in a general Japanese population: the Hisayama study. AB - OBJECTIVE: Several studies have shown associations between diabetes and various types of cancer other than gastric cancer. The aim of this cohort study was to evaluate the impact of fasting plasma glucose (FPG) levels on gastric cancer occurrence. RESEARCH DESIGN AND METHODS: A total of 2,466 Japanese subjects aged > or =40 years were stratified into three groups according to FPG tertiles (<5.3 mmol/l, low FPG; 5.3-5.8 mmol/l, modest FPG; >5.8 mmol/l, high FPG) and followed up prospectively for 9 years. RESULTS: During the follow-up, 66 subjects experienced gastric cancer. In men, the age-adjusted incidences were significantly higher in the modest-FPG (7.0 per 1,000 person-years, P < 0.05) and high-FPG (7.2, P < 0.05) groups than in the low-FPG group (2.2). In women, the high-FPG group also had a significantly higher age-adjusted incidence of gastric cancer compared with the low-FPG group (2.5 vs. 0.8, P < 0.05). The multivariate analysis with Cox's proportional hazards model revealed that the risks of gastric cancer in the modest-FPG (relative risk [RR] 2.3 [95% CI 1.1-5.0]) and high-FPG (3.1 [1.5-6.4]) groups were significantly higher than that in the low-FPG group, even after adjusting for other comprehensive risk factors, including Helicobacter pylori status, smoking, and dietary factors. However, this FPG-cancer association was observed only among H. pylori-seropositive subjects. CONCLUSIONS: Our findings suggest that a modest increase in FPG is a risk factor for gastric cancer and that hyperglycemia is a possible cofactor increasing the risk posed by Helicobacter pylori infection. PMID- 15793175 TI - The prevalence of diabetes and impaired glucose tolerance in Sivas, Central Anatolia, Turkey. AB - OBJECTIVE: The aim of this study was to determine type 2 diabetes, impaired glucose tolerance (IGT), and impaired fasting glucose (IFG) prevalence in Sivas, Turkey. RESEARCH DESIGN AND METHODS: This cross-sectional study was conducted in the city center of Sivas. The study population of 771 subjects was selected by the cluster sampling method from 115,998 individuals aged > or =30 years. Participants with fasting venous plasma glucose concentrations <100 mg/dl were classified as "normal." Diabetes was diagnosed in participants if they had fasting blood glucose levels > or =126 mg/dl. An oral glucose tolerance test (OGTT) was performed in subjects with fasting blood glucose levels > or =100 mg/dl and <126 mg/dl. RESULTS: According to the fasting blood glucose levels of the 771 subjects, 44 (5.7%) had diabetes. OGTTs were performed in 80 (10.4%) subjects. According to OGTT results, there were 5 subjects with diabetes, 20 subjects with IGT (2.6%), and 55 subjects with IFG (7.1%). The combined prevalence of IFG and IGT was 9.7%. After OGTT, the total number of diabetic subjects was determined to be 49 (6.4%). Twenty-four (3.1%) of the subjects had a previous diagnosis of diabetes. Multivariate analyses showed that age, sex, hypertension, cigarette smoking, obesity, and family history of diabetes were risk factors for type 2 diabetes (P < 0.05). CONCLUSIONS: Diabetes incidence increases with changes in dietary habits and lifestyle. Education is particularly important for public health, as the community may then have required knowledge about the disease and its risk factors. PMID- 15793176 TI - Physical activity, cardiovascular risk factors, and mortality among Finnish adults with diabetes. AB - OBJECTIVE: The aim of this study was to examine both single and joint associations of physical activity and conventional cardiovascular risk factors with total and cardiovascular mortality among patients with diabetes. RESEARCH DESIGN AND METHODS: We prospectively followed 3,708 Finnish patients with type 2 diabetes aged 25-74 years. Physical activity, smoking status, blood pressure, height, weight, and serum cholesterol level were determined at baseline. Cox proportional hazard models were used to estimate single and joint effects of physical activity and other cardiovascular risk factors on the risk of mortality. RESULTS: During a mean follow-up of 18.7 years, 1,423 deaths were recorded, 906 of which were due to cardiovascular disease. Moderate or high levels of physical activity were associated with decreased total and cardiovascular mortality, whereas higher levels of BMI and blood pressure and current smoking were associated with increased total and cardiovascular mortality. High serum cholesterol levels also increased cardiovascular mortality. The protective effect of physical activity was consistent in diabetic patients with any levels of BMI, blood pressure, total cholesterol, and smoking. CONCLUSIONS: A moderate or high level of physical activity was associated with a reduced risk of total and cardiovascular mortality among patients with type 2 diabetes. The favorable association of physical activity with longevity was observed regardless of the levels of BMI, blood pressure, total cholesterol, and smoking. PMID- 15793177 TI - Use of HLA typing in diagnosing celiac disease in patients with type 1 diabetes. AB - OBJECTIVE: This study examines the use of HLA typing for the diagnosis of celiac disease in a group of Australians with type 1 diabetes. RESEARCH DESIGN AND METHODS: Subjects included 131 sequential patients with type 1 diabetes (mean age 17 years [range 10-37]), 77 patients with biopsy-proven celiac disease (mean age 52 years [range 12-84]), and 162 healthy control subjects (mean age 17 years [range 2 months to 56 years]). Subjects were prospectively screened for celiac disease using endomysial antibodies (EMAs), tissue transglutaminase antibodies (TTGAs), and celiac disease-specific HLA typing. RESULTS: Celiac disease was diagnosed in 11 subjects after an intestinal biopsy (prevalence 8.4%). There was 95% agreement between TTGA and EMA for positive results and 100% for negative results. There was no significant difference for HLA DQ2 and DR4 among patients with type 1 diabetes with or without celiac disease. CONCLUSIONS: The prevalence of celiac disease among patients with type 1 diabetes is higher than previously estimated in Australia. TTGA is a valuable diagnostic tool that can be used for screening celiac disease in patients with type 1 diabetes. HLA typing should not be used in the diagnosis of celiac disease in patients with type 1 diabetes because of the similarities of HLA types between patients with type 1 diabetes and those with celiac disease. PMID- 15793178 TI - The relation between hyperglycemia and outcomes in 2,471 patients admitted to the hospital with community-acquired pneumonia. AB - OBJECTIVE: To examine whether hyperglycemia at the time of presentation was associated with outcomes in patients admitted to non-intensive care settings with community-acquired pneumonia (CAP). RESEARCH DESIGN AND METHODS: Prospective cohort study of consecutive patients admitted to six hospitals between 15 November 2000 and 14 November 2002. RESULTS: Of the 2,471 patients in this study (median age 75 years), 279 (11%) had serum glucose at presentation >11 mmol/l: 178 of the 401 patients (44%) with a prior diagnosis of diabetes and 101 of the 2,070 patients (5%) without a history of diabetes. Of patients hospitalized with CAP, 9% died and 23% suffered an in-hospital complication. Compared with those with values < or =11 mmol/l, patients with an admission glucose >11 mmol/l had an increased risk of death (13 vs. 9%, P = 0.03) and in-hospital complications (29 vs. 22%, P = 0.01). Compared with those patients with admission glucose < or =6.1 mmol/l, the mortality risk was 73% higher (95% CI 12-168%) and the in-hospital complication risk was 52% higher (12-108%) in patients with admission glucose >11 mmol/l. Even after adjustment for factors in the Pneumonia Severity Index, hyperglycemia on admission remained significantly associated with subsequent adverse outcomes: for each 1-mmol/l increase, risk of in-hospital complications increased 3% (0.2-6%). CONCLUSIONS: Hyperglycemia on admission is independently associated with adverse outcomes in patients with CAP, with the increased risks evident at lower glucose levels than previously reported. PMID- 15793179 TI - The relationship between knowledge of recent HbA1c values and diabetes care understanding and self-management. AB - OBJECTIVE: Knowledge of one's actual and target health outcomes (such as HbA(1c) values) is hypothesized to be a prerequisite for effective patient involvement in managing chronic diseases such as diabetes. We examined 1) the frequency and correlates of knowing one's most recent HbA(1c) test result and 2) whether knowing one's HbA(1c) value is associated with a more accurate assessment of diabetes control and better diabetes self-care understanding, self-efficacy, and behaviors related to glycemic control. RESEARCH DESIGN AND METHODS: We conducted a cross-sectional survey of a sample of 686 U.S. adults with type 2 diabetes in five health systems who had HbA(1c) checked in the previous 6 months. Independent variables included patient characteristics, health care provider communication, and health system type. We examined bivariate and multivariate associations between each variable and the respondents' knowledge of their last HbA(1c) values and assessed whether knowledge of HbA(1c) was associated with key diabetes care attitudes and behaviors. RESULTS: Of the respondents, 66% reported that they did not know their last HbA(1c) value and only 25% accurately reported that value. In multivariate analyses, more years of formal education and high evaluations of provider thoroughness of communication were independently associated with HbA(1c) knowledge. Respondents who knew their last HbA(1c) value had higher odds of accurately assessing their diabetes control (adjusted odds ratio 1.59, 95% CI 1.05-2.42) and better reported understanding of their diabetes care (P < 0.001). HbA(1c) knowledge was not associated with respondents' diabetes care self efficacy or reported self-management behaviors. CONCLUSIONS: Respondents who knew their HbA(1c) values reported better diabetes care understanding and assessment of their glycemic control than those who did not. Knowledge of one's HbA(1c) level alone, however, was not sufficient to translate increased understanding of diabetes care into the increased confidence and motivation necessary to improve patients' diabetes self-management. Strategies to provide information to patients must be combined with other behavioral strategies to motivate and help patients effectively manage their diabetes. PMID- 15793180 TI - Prevalence and correlates of depressive symptoms among rural older African Americans, Native Americans, and whites with diabetes. AB - OBJECTIVE: Depression is associated with morbidity, mortality, and decreased quality of life and is a well-established complication among people with diabetes. Little is known about the prevalence and correlates of depressive symptoms among older adults living in rural communities, particularly among ethnic minority groups, who are at increased risk of developing diabetes and complications. RESEARCH DESIGN AND METHODS: Data were analyzed from the ELDER (Evaluating Long-term Diabetes Self-management Among Elder Rural Adults) diabetes study in which face-to-face interviews were conducted with 696 older (> or =65 years of age) African-American, Native American, and white men and women in two rural counties in central North Carolina. RESULTS: Using a criterion of > or =9 on a modified CES-D (Center for Epidemiologic Study of Depression) scale, 15.8% of the sample had depressive symptoms. In bivariate analyses, depressive symptomatology was more common among women and individuals who were unmarried and had less than a high school education, fewer financial resources, more chronic conditions, more prescription medications, and lower physical functioning. In multivariate analyses, sex, education, living arrangement, BMI, number of prescription medications, number of chronic conditions, and physical functioning remained significant. CONCLUSIONS: These results show that older rural adults with diabetes are at high risk for depressive symptoms, regardless of their ethnic group, and that certain demographic and health characteristics are important factors in this association. These findings add to the limited body of knowledge of comorbid depression in this population. Greater attention should be paid to diagnosing and treating this condition by those who provide care to these populations. PMID- 15793181 TI - Depression symptoms and antidepressant medicine use in Diabetes Prevention Program participants. AB - OBJECTIVE: To assess depression markers (symptoms and antidepressant medicine use) in Diabetes Prevention Program (DPP) participants and to determine whether changes in depression markers during the course of the study were associated with treatment arm, weight change, physical activity level, or participant demographic characteristics. RESEARCH DESIGN AND METHODS: DPP participants (n = 3,187) in three treatment arms (intensive lifestyle, metformin, and placebo) completed the Beck Depression Inventory (BDI) and reported on use of antidepressant medicines at randomization and subsequently at each annual visit (average duration in study 3.2 years). RESULTS: On study entry, 10.3% of participants had BDI scores > or =11, which was used as a threshold for mild depression, 5.7% took antidepressant medicines, and 0.9% had both depression markers. During the DPP, the proportion of participants with elevated BDI scores declined (from 10.3% at baseline to 8.4% at year 3), while the proportion taking antidepressant medicines increased (from 5.7% at baseline to 8.7% at year 3), leaving the proportion with either marker unchanged. These time trends were not significantly associated with the DPP treatment arm. Depression markers throughout the study were associated with some participant demographic factors, adjusted for other factors. Men were less likely to have elevated depression scores and less likely to use antidepressant medicine at baseline (9.0% of men and 17.9% of women had at least one marker of depression) and throughout the study (P <0.0001). Those with more education were less likely to have elevated symptom scores (P = 0.0007) but more likely to be taking antidepressant medicine (P = 0.002). Non-Hispanic white participants were less likely than African Americans to have BDI scores > or =11 (P = 0.03), but white participants were more likely to be taking antidepressant medicine than any other racial/ethnic group (P <0.0001). CONCLUSIONS: DPP participation was not associated with changes in levels of depression. Countervailing trends in the proportion of DPP participants with elevated depression symptoms and the proportion taking antidepressant medicine resulted in no significant change in the proportion with either marker. The finding that those taking antidepressant medicine often do not have elevated depression symptoms indicates the value of assessing both markers when estimating overall depression rates. PMID- 15793182 TI - Estimation of glomerular filtration rate in diabetic subjects: Cockcroft formula or modification of Diet in Renal Disease study equation? AB - OBJECTIVE: The Cockcroft-Gault formula is recommended for the evaluation of renal function in diabetic patients. The more recent Modification of Diet in Renal Disease (MDRD) study equation seems more accurate, but it has not been validated in diabetic patients. This study compares the two methods. RESEARCH DESIGN AND METHODS: In 160 diabetic patients, we compared the Cockcroft-Gault formula and MDRD equation estimations to glomerular filtration rates (GFRs) measured by an isotopic method ((51)Cr-EDTA) by correlation studies and a Bland-Altman procedure. Their accuracy for the diagnosis of moderately (GFR <60 ml . min(-1) . 1.73 m(-2)) or severely (GFR <30 ml . min(-1) . 1.73 m(-2)) impaired renal function were compared with receiver operating characteristic (ROC) curves. RESULTS: Both the Cockcroft-Gault formula (r = 0.74; P < 0.0001) and MDRD equation (r = 0.81; P < 0.0001) were well correlated with isotopic GFR. The Bland Altman procedure revealed a bias for the MDRD equation, which was not the case for the Cockcroft-Gault formula. Analysis of ROC curves showed that the MDRD equation had a better maximal accuracy for the diagnosis of moderate (areas under the curve [AUCs] 0.868 for the Cockcroft-Gault formula and 0.927 for the MDRD equation; P = 0.012) and severe renal failure (AUC 0.883 for the Cockcroft-Gault formula and 0.962 for the MDRD equation; P = 0.0001). In the 87 patients with renal insufficiency, the MDRD equation estimation was better correlated with isotopic GFR (Cockcroft-Gault formula r = 0.57; the MDRD equation r = 0.78; P < 0.01), and it was not biased as evaluated by the Bland-Altman procedure. CONCLUSIONS: Although both equations have imperfections, the MDRD equation is more accurate for the diagnosis and stratification of renal failure in diabetic patients. PMID- 15793183 TI - Rosiglitazone improves postprandial triglyceride and free fatty acid metabolism in type 2 diabetes. AB - OBJECTIVE: Increased postprandial lipemia is part of diabetic dyslipidemia and is associated with accelerated atherosclerosis. We investigated the effects of the peroxisome proliferator-activated receptor-gamma agonist rosiglitazone on postprandial lipemia in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: A randomized, 8-week, crossover, placebo-controlled, double-blind trial was performed in which rosiglitazone at 4 mg was administrated twice daily in 19 patients with type 2 diabetes. Standardized 6-h oral fat-loading tests were performed after each treatment period. Postprandial curves were calculated as the total area under the curve (AUC) and the incremental area under the curve (dAUC). RESULTS: Rosiglitazone did not change fasting plasma triglycerides compared with placebo (1.97 +/- 0.22 vs. 1.88 +/- 0.20 mmol/l, respectively) but decreased postprandial triglyceride levels, leading to significantly lower triglyceride dAUC (-37%, P < 0.05), without changing total triglyceride AUC. Significant postprandial triglyceride reductions in the chylomicron fraction (Svedberg flotation rate [Sf] >400) were achieved with rosiglitazone, which resulted in a significant lower triglyceride AUC (-22%) in this fraction. The postprandial triglyceride increase in VLDL1 (Sf 60-400) was also lower after rosiglitazone ( 27%), but this did not result in a significant lower triglyceride AUC. In VLDL2 (Sf 20-60), there were no significant differences in triglyceride AUC and triglyceride dAUC between rosiglitazone and placebo. Rosiglitazone decreased free fatty acid (FFA) AUC (-12%) and FFA dAUC (-18%) compared with placebo. CONCLUSIONS: Rosiglitazone improves the metabolism of large triglyceride-rich lipoproteins and decreases postprandial FFA concentrations in type 2 diabetes. This may have clinical implications, as these effects may contribute to cardiovascular risk reduction. PMID- 15793184 TI - Autoantibody "subspecificity" in type 1 diabetes: risk for organ-specific autoimmunity clusters in distinct groups. AB - OBJECTIVE: Autoimmune thyroid disease (AIT), celiac disease, and Addison's disease are characterized by the presence of autoantibodies: thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) in AIT, tissue transglutaminase antibody (TTGAb) in celiac disease, and 21-hydroxylase antibody (21-OHAb) in Addison's disease. The objective of this study was to define the prevalence of these autoantibodies and clinical disease in a population with type 1 diabetes. RESEARCH DESIGN AND METHODS: We screened 814 individuals with type 1 diabetes for TPOAb, TGAb, TTGAb, and 21-OHAb. Clinical disease was defined by chart review. Factors related to the presence of autoimmunity and clinical disease including age at onset of type 1 diabetes, duration of diabetes, age at screening, sex, and the presence of autoantibodies were reviewed. RESULTS: The most common autoantibodies expressed were TPOAb and/or TGAb (29%), followed by TTGAb (10.1%) and 21-OHAb (1.6%). Specific HLA DR/DQ genotypes were associated with the highest risk for expression of 21-OHAb (DRB1*0404-DQ8, DR3-DQ2) and TTGAb (DR3-DQ2- DR3-DQ2). The expression of thyroid autoantibodies was related to 21-OHAb but not to TTGAb. The presence of autoantibodies was associated with and predictive of disease. CONCLUSIONS: In this large cohort of individuals with type 1 diabetes, the expression of organ-specific autoantibodies was very high. The grouping of autoantibody expression suggests common factors contributing to the clustering. PMID- 15793185 TI - Effect of maternal use of antiretroviral agents on serum insulin levels of the newborn infant. AB - OBJECTIVE: The aim of this study was to investigate the effect of antiretroviral drugs on neonatal serum insulin levels. RESEARCH DESIGN AND METHODS: A prospective study was conducted on 57 pregnant women divided into three groups: the zidovudine (ZDV) group, HIV-infected women taking ZDV (n = 20); the triple treatment group, HIV-infected women taking triple antiretroviral agents ZDV + lamivudine + nelfinavir (n = 25); and the control group, pregnant women considered normal from a clinical and laboratory standpoint (n = 12). Blood was collected from the umbilical cord of newborn infants upon delivery for measurement of insulin level. The insulin measurements were performed in duplicate by radioimmunoassay. RESULTS: Demographic and anthropometric data were homogeneous, and pregnant women with a personal and family history of diabetes were excluded. There was no difference between groups regarding glycemia in the newborn. Median newborn insulin doses were 2.9, 4.8, and 6.5 muU/ml for the triple treatment, ZDV, and control groups, respectively (P < 0.05). CONCLUSIONS: Use of triple therapy during pregnancy induced a significant decrease in serum levels of neonatal insulin compared with the control group. Active surveillance of short- and long-term adverse events is imperative to issue a definitive statement regarding the impact that use of protease inhibitors during pregnancy will have on infant metabolism. PMID- 15793186 TI - Proinsulin concentration is an independent predictor of all-cause and cardiovascular mortality: an 11-year follow-up of the Hoorn Study. AB - OBJECTIVE: High proinsulin concentration may be a better predictor for cardiovascular disease (CVD) mortality than insulin concentration. Previous observations may have been confounded by glucose tolerance status or lack of precision because of high intraindividual variability. We investigated the longitudinal relation of means of duplicate measurements of insulin and proinsulin with all-cause and CVD mortality in a population-based cohort taking glucose tolerance status into account. RESEARCH DESIGN AND METHODS: Fasting and post-75-g glucose-load (2-h) glucose, insulin, and proinsulin values were determined in duplicate on separate days in 277 participants with normal glucose metabolism, 208 participants with impaired glucose metabolism, and 119 newly detected patients with type 2 diabetes of the Hoorn Study. Insulin resistance and beta-cell function were estimated by homeostasis model assessment (HOMA-IR and HOMA-B, respectively), and the fasting proinsulin-to-insulin ratio was calculated. Subjects were followed with respect to mortality until January 2003. RESULTS: Fasting proinsulin levels were significantly associated with all-cause and CVD mortality. The hazard ratios (HRs) per increase in interquartile range adjusted for age and sex were 1.21 (95% CI 1.04-1.42) for all-cause mortality and 1.33 (1.06-1.66) for CVD mortality. Adjustment for glucose tolerance status and HOMA-IR did not substantially change the associations. CONCLUSIONS: Fasting proinsulin was associated with all-cause and CVD mortality, independent of glucose tolerance status and insulin resistance and largely independent of other CVD risk factors. Proinsulin might play a role in the relationship between insulin resistance and CVD. PMID- 15793187 TI - Coronary artery disease is common in nonuremic, asymptomatic type 1 diabetic islet transplant candidates. AB - OBJECTIVE: Coronary artery disease (CAD) is the most common cause of death in patients with type 1 diabetes. Asymptomatic CAD is common in uremic diabetic patients, but its prevalence in nonuremic type 1 diabetic patients is unknown. The prevalence of CAD was determined by coronary angiography and the performance of noninvasive cardiac investigation evaluated in type 1 diabetic islet transplant (ITX) candidates with preserved renal function. RESEARCH DESIGN AND METHODS: A total of 60 consecutive type 1 diabetic ITX candidates (average age 46 years [mean 24-64], 23 men, and 47% ever smokers) underwent coronary angiography, electrocardiographic stress testing (EST), and myocardial perfusion imaging (MPI) in a prospective cohort study. CAD was indicated on angiography by the presence of stenoses >50%. Models to predict CAD were examined by logistic regression. RESULTS: Most subjects (53 of 60) had no history or symptoms of CAD; 23 (43%) of these asymptomatic subjects had stenoses >50%. CAD was associated with age, duration of diabetes, hypertension, and smoking. Although specific, EST and MPI were not sensitive as predictors of CAD on angiography (specificity 0.97 and 0.93, sensitivity 0.17 and 0.04, respectively) but helped identify two of three subjects requiring revascularization. EST and MPI did not enhance logistic regression models. A clinical algorithm to identify low-risk subjects who may not require angiography was highly sensitive but was applicable only to a minority (n = 8, sensitivity 1.0, specificity 0.27, negative predictive value 1.0). CONCLUSIONS: Nonuremic type 1 diabetic patients with hypoglycemic unawareness and/or metabolic lability referred for ITX are at high risk for asymptomatic CAD despite negative noninvasive investigations. Aggressive management of cardiovascular risk factors and further investigation into optimal cardiac risk stratification in type 1 diabetes are warranted. PMID- 15793188 TI - Metabolic syndrome and risk of restenosis in patients undergoing percutaneous coronary intervention. AB - OBJECTIVE: Patients with metabolic syndrome have increased risk of cardiovascular events. The number of patients with metabolic syndrome is rapidly increasing, and these patients often need revascularization. However, only limited data are available on the effect of metabolic syndrome on restenosis in patients undergoing percutaneous coronary intervention (PCI). RESEARCH DESIGN AND METHODS: To assess the role of metabolic syndrome in the development of restenosis, we performed an analysis in a population of patients from the GENetic DEterminants of Restenosis (GENDER) study. The GENDER project, a multicenter prospective study, included consecutive patients after successful PCI and was designed to study the predictive value of various genetic and other risk factors for subsequent clinical restenosis, defined as target vessel revascularization (TVR) or combined end point of death, myocardial infarction, and TVR. This subpopulation of GENDER consisted of 901 patients, 448 of whom (49.7%) had metabolic syndrome. RESULTS: On multivariable Cox regression analysis, controlling for age, sex, previous myocardial infarction, stent length, current smoking, and statin therapy, there was no association between increased risk of TVR (hazard ratio 1.03 [95% CI 0.68-1.57]) or the combined end point (1.05 [0.71 1.55]) and the presence of metabolic syndrome. CONCLUSIONS: This study demonstrates that metabolic syndrome is not associated with TVR or the combined end point after PCI. Furthermore, accumulating characteristics of metabolic syndrome were neither associated with increased risk of TVR nor with the combined end point. Therefore, PCI has equal beneficial results in patients with or without metabolic syndrome. This is important information in light of the pandemic proportion of metabolic syndrome that the medical community will face. PMID- 15793189 TI - The metabolic syndrome and concentrations of C-reactive protein among U.S. youth. AB - OBJECTIVE: Adults with the metabolic syndrome show biochemical evidence of low grade inflammation. We sought to examine whether this is true among U.S. youth with the metabolic syndrome. RESEARCH DESIGN AND METHODS: We used data from 1,366 participants aged 12-17 years from the National Health and Nutrition Examination Survey 1999-2000. A modification of the definition of the metabolic syndrome proposed by the Third Report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults was used. C-reactive protein (CRP) was measured by latex-enhanced nephelometry. RESULTS: Mean and median concentrations of CRP were higher among participants who had the metabolic syndrome (mean 3.8 mg/l, geometric mean 1.8 mg/l) than among those who did not (mean 1.4 mg/l, geometric mean 0.4 mg/l). The percentage of participants with a concentration of CRP >3.0 mg/l was 38.4% among those with the metabolic syndrome and 10.3% among those without the syndrome (P = 0.007). Of the five components of the syndrome, only abdominal obesity was significantly and independently associated with log-transformed concentrations of CRP in multiple linear regression analysis. CONCLUSIONS: Our results show that a large percentage of children and adolescents with the metabolic syndrome have elevated concentrations of CRP. Whether the elevated concentrations of CRP among children and adolescents who have the metabolic syndrome predict future adverse health events remains to be determined. PMID- 15793190 TI - The metabolic syndrome in older individuals: prevalence and prediction of cardiovascular events: the Cardiovascular Health Study. AB - OBJECTIVE: The prevalence of the metabolic syndrome, a potent risk factor for cardiovascular diseases (CVDs), has not been adequately explored in older individuals. Moreover, two sets of criteria have been proposed for the definition of metabolic syndrome, one by the World Health Organization (WHO) and one by the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATPIII). We therefore investigated the prevalence of this syndrome in a subgroup of older participants from the Cardiovascular Health Study (CHS) who were free of CVD at baseline. We also compared the prognostic significance of the two definitions of the metabolic syndrome. RESEARCH DESIGN AND METHODS: A total of 2,175 subjects from the CHS who were free of CVD at baseline and not taking antihypertensive or lipid-lowering medications were studied. Prevalence of the metabolic syndrome was assessed with both the WHO and ATPIII criteria. The incidence of coronary or cerebrovascular disease was ascertained during a median follow-up time of 4.1 years. RESULTS: Prevalence of the metabolic syndrome was 28.1% by ATPIII criteria and 21.0% by WHO criteria. The two sets of criteria provided concordant classification for 80.6% of participants. Multivariate Cox propotional hazard models showed that the metabolic syndrome defined with the ATPIII criteria, but not with the WHO criteria, was an independent predictor of coronary or cerebrovascular events and was associated with a 38% increased risk (hazard ratio 1.38 [95% CI 1.06-1.79], P < 0.01). CONCLUSIONS: Prevalence of the metabolic syndrome in older individuals is approximately 21-28% (depending on the definition used). The two sets of criteria have 80% concordance in classifying subjects. As defined by the ATPIII criteria, the metabolic syndrome yields independent prognostic information, even after adjusting for traditional cardiovascular risk factors and the individual domains of the metabolic syndrome. PMID- 15793191 TI - Impact of intensive lifestyle and metformin therapy on cardiovascular disease risk factors in the diabetes prevention program. AB - OBJECTIVE: The Diabetes Prevention Program demonstrated the ability to delay or prevent type 2 diabetes in participants with impaired glucose tolerance (IGT). Participants with IGT are at high risk for cardiovascular disease (CVD), with a marked increase in the number and severity of CVD risk factors. We prospectively assessed the impact of our interventions on hypertension, dyslipidemia, and CVD events. RESEARCH DESIGN AND METHODS: The study group consisted of 3,234 individuals with IGT randomly assigned to receive intensive lifestyle intervention, metformin, or placebo. Annual assessment of blood pressure, lipids, electrocardiogram, and CVD events was undertaken. RESULTS: Hypertension was present in 30% of participants at study entry and then increased in the placebo and metformin groups, although it significantly decreased with intensive lifestyle intervention. Triglyceride levels fell in all treatment groups, but fell significantly more with intensive lifestyle intervention. Total cholesterol and LDL cholesterol levels were similar among treatment groups. Intensive lifestyle intervention significantly increased the HDL cholesterol level and reduced the cumulative incidence of the proatherogenic LDL phenotype B. At 3 years of follow-up, the use for pharmacologic therapy to achieve established goals in the intensive lifestyle group was 27-28% less for hypertension and 25% less for hyperlipidemia compared with placebo and metformin groups. Over an average of 3 years, 89 CVD events from 64 participants were positively adjudicated studywide, with no differences among treatment groups. CONCLUSIONS: Lifestyle intervention improves CVD risk factor status compared with placebo and metformin therapy. Although no differences in CVD events were noted after 3 years, achieved risk factor modifications suggest that longer intervention may reduce CVD event rates. PMID- 15793192 TI - Cardiorespiratory fitness attenuates metabolic risk independent of abdominal subcutaneous and visceral fat in men. AB - OBJECTIVE: Moderate to high levels of cardiorespiratory fitness (CRF) are associated with a lower risk of the metabolic syndrome and all-cause mortality. Unknown is whether CRF attenuates health risk for a given level of abdominal visceral fat, subcutaneous fat, and/or waist circumference. RESEARCH DESIGN AND METHODS: The sample studied comprised 297 apparently healthy men with available computed tomography or magnetic resonance imaging scans of the abdomen, metabolic data, and maximal treadmill exercise test results. Men were categorized into low CRF (20%, n = 56), moderate-CRF (40%, n = 94), and high-CRF (40%, n = 147) groups based on age and exercise test results. All analyses were adjusted for age. RESULTS: For a given level of waist circumference, visceral fat, or subcutaneous fat, the high-CRF group had lower triglyceride levels (P < 0.05) and higher HDL cholesterol levels than the low- or moderate-CRF groups. There was a significant group interaction (P < 0.01) for blood pressure, indicating that the increase in blood pressure per unit increase in visceral fat or waist circumference was greater in men in the low-CRF group compared with the high-CRF group. The relative risks of having the metabolic syndrome were 1.8 (95% CI 1.0-3.1) and 1.6 (0.9-2.7) times higher in the low- and moderate-CRF groups, respectively, compared with the high-CRF group after adjusting for age, visceral fat, and subcutaneous fat (P for trend = 0.06). CONCLUSIONS: High levels of CRF are associated with a substantial reduction in health risk for a given level of visceral and subcutaneous fat. PMID- 15793193 TI - Predictors of changes in glucose tolerance status in obese youth. AB - OBJECTIVE: Type 2 diabetes in obese youth is an emerging problem. The metabolic and anthropometric predictors of change in glucose tolerance status in obese youth are unknown. RESEARCH DESIGN AND METHODS: A total of 117 obese children and adolescents were studied by performing an oral glucose tolerance test (OGTT) at baseline and after approximately 2 years. Data from both OGTTs and changes in weight were examined to identify youth at highest risk for developing diabetes and the factors that have the strongest impact on glucose tolerance. RESULTS: Eighty-four subjects had normal glucose tolerance (NGT) and 33 impaired glucose tolerance (IGT) at baseline. Eight subjects (all of whom had IGT at baseline) developed type 2 diabetes, whereas 15 subjects with IGT reverted to NGT. In this cohort, severe obesity, impaired glucose tolerance, and African-American background emerged as the best predictors of developing type 2 diabetes, whereas fasting glucose, insulin, and C-peptide were nonpredictive. Changes in insulin sensitivity, strongly related to weight change, had a significant impact on the 2 h glucose level on the follow-up study. CONCLUSIONS: Severely obese children and adolescents with IGT, particularly of African-American descent, are at very high risk for developing type 2 diabetes over a short period of time. Parameters derived from an OGTT and not fasting samples can serve as predictors of changes in glucose tolerance. PMID- 15793194 TI - Increased urinary albumin excretion, insulin resistance, and related cardiovascular risk factors in patients with type 2 diabetes: evidence of a sex specific association. AB - OBJECTIVE: While the relevant role of insulin resistance in the pathogenesis of increased urinary albumin excretion (UAE) is well established in type 1 diabetes, its contribution in type 2 diabetes is controversial. Our aim was to investigate whether insulin resistance was associated with increased UAE in a large cohort of patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 363 men and 349 women, aged 61 +/- 9 years, with a disease duration of 11 +/- 9 years and HbA(1c) levels of 8.6 +/- 2.0% were included. Insulin resistance was derived from the homeostasis model assessment of insulin resistance (HOMA(IR)), and UAE was derived from the albumin-to-creatinine ratio (ACR) defined as increased if the value was > or =2.5 mg/mmol in men and > or =3.5 mg/mmol in women. ACR was correlated with HOMA(IR) (r = 0.15, P = 0.0001), independently of age, disease duration, blood pressure, HbA(1c), triglycerides, waist circumference, and smoking. RESULTS: When the two sexes were investigated separately, a significant correlation between ACR and HOMA(IR) was reached in men (n = 363; r = 0.21, P = 0.0001) but not women (n = 349; r = 0.08, P = 0.14), suggesting that insulin resistance and sex may interact (P for interaction = 0.04) in determining UAE. When men were subgrouped into quartiles of HOMA(IR), those of the third and fourth quartile (i.e., the most insulin resistant) were at higher risk to have increased ACR than patients of the first quartile (third quartile: odds ratio 2.2 [95% CI 1.2-4.2], P = 0.01) (fourth quartile: 4.1 [2.2-7.9], P = 0.00002). Finally, ACR was significantly higher in men with two or more insulin resistance related cardiovascular risk factors (i.e., abdominal obesity, dyslipidemia, and arterial hypertension) than in men with fewer than two insulin resistance-related cardiovascular risk factors (0.90 [0.2-115.1] vs. 1.56 [0.1-1367.6], respectively, P = 0.005). CONCLUSIONS: In type 2 diabetic patients, increased UAE is strongly associated with insulin resistance and related cardiovascular risk factors. This association seems to be stronger in men than in women. PMID- 15793195 TI - Maternal efforts to prevent type 1 diabetes in at-risk children. AB - OBJECTIVE: The aim of this study was to assess maternal diabetes prevention efforts aimed at children identified as at risk through newborn genetic screening. RESEARCH DESIGN AND METHODS: A total of 192 mothers of children identified as at risk for type 1 diabetes through newborn genetic screening were administered a structured interview 3.6 +/- 0.8 years after risk notification. The interview assessed possible diabetes prevention behaviors across six domains: health surveillance, diet, physical activity, illness prevention, medications, and stress reduction. A mother's cognitive (diabetes risk perception and perceived control), affective (anxiety), and coping responses to the child's at risk status were assessed. RESULTS: A total of 67% of mothers reported one or more diabetes prevention behaviors. Monitoring behaviors (e.g., watching for signs of diabetes and checking blood glucose) were the most common, reported in 59%, followed by modifications in the child's diet in 34% and physical activity in 14%. Potentially harmful prevention behaviors (e.g., limiting contact with other children, delaying immunizations, and giving medications including insulin) were rare. Mothers who engaged in diabetes prevention behaviors reported higher diabetes risk perception, greater anxiety, and more use of certain coping styles. Infants of these mothers were more likely to have a first-degree relative with diabetes. CONCLUSIONS: In the absence of known methods of preventing type 1 diabetes, most mothers of at-risk children report diabetes prevention behaviors. Such behaviors must be more carefully assessed to ensure accurate interpretation of data obtained from natural history studies and prevention trials. PMID- 15793196 TI - Renal status among patients using metformin in a primary care setting. PMID- 15793197 TI - Type 1 diabetes and autism: is there a link? PMID- 15793198 TI - Low incidence of type 1 diabetes in Iran. PMID- 15793199 TI - Revisiting the total contact cast: maximizing off-loading by wound isolation. PMID- 15793200 TI - Lipoprotein(a) as a risk factor for cardiovascular mortality in type 2 diabetic patients: a 10-year follow-up study. PMID- 15793202 TI - Left ventricular mass in patients with type 2 diabetes is independently associated with central but not peripheral pulse pressure. PMID- 15793201 TI - SDF-1 genotype influences insulin-dependent mobilization of adult progenitor cells in type 2 diabetes. PMID- 15793203 TI - Hepatitis C virus infection and human pancreatic beta-cell dysfunction. PMID- 15793204 TI - Effect of sibutramine on weight management and metabolic control in type 2 diabetes: a meta-analysis of clinical studies. AB - OBJECTIVE: The aim of this study was to provide a comprehensive meta-analysis of randomized controlled clinical studies on the effects of sibutramine on weight loss and glycemic control in obese subjects with type 2 diabetes. RESEARCH DESIGN AND METHODS: Controlled clinical trials assessing the effect sizes of sibutramine on weight loss effects on glycemia in obese subjects with type 2 diabetes were identified and reviewed using the Cochrane Library, Medline, EMBASE, and a manual search. RESULTS: Eight placebo-controlled, double-blind, randomized trials of sibutramine were included. After sibutramine treatment, the decrease in body weight and waist circumference was significantly greater than in the placebo group. Fasting blood glucose and HbA(1c) significantly decreased after sibutramine treatment. Treatment benefits were seen in plasma triglycerides and HDL, without significant variations in serum total and LDL cholesterol. No differences in systolic blood pressure between the sibutramine and the placebo groups were seen, while recording of diastolic blood pressure and heart rate showed that sibutramine produced a small increase relative to placebo. CONCLUSIONS: A pharmacological approach in a weight management program for patients with type 2 diabetes may be helpful in glycemic control and in the management of other risk factors. Sibutramine may help improve glucose control because it is conducive to weight loss. The reviewed data on the effect of sibutramine further enforce the recommendations that weight management may be the most important therapeutic task for most obese subjects with type 2 diabetes. PMID- 15793205 TI - Reduced hypoglycemia risk with insulin glargine: a meta-analysis comparing insulin glargine with human NPH insulin in type 2 diabetes. AB - OBJECTIVE: Insulin glargine (LANTUS) is a once-daily basal insulin analog with a smooth 24-h time-action profile that provides effective glycemic control with reduced hypoglycemia risk (particularly nocturnal) compared with NPH insulin in patients with type 2 diabetes. A recent "treat-to-target" study has shown that more patients on insulin glargine reached HbA(1c) levels < or =7.0% without confirmed nocturnal hypoglycemia compared with NPH insulin. We further assessed the risk for hypoglycemia in a meta-analysis of controlled trials of a similar design for insulin glargine versus once- or twice-daily NPH insulin in adults with type 2 diabetes. RESEARCH DESIGN AND METHODS: All studies were 24-28 weeks long, except one 52-week study, for which interim 20-week data were used. RESULTS: Patient demographics were similar between the insulin glargine (n = 1,142) and NPH insulin (n = 1,162) groups. The proportion of patients achieving target HbA(1c) (< or =7.0%) was similar between insulin glargine-and NPH insulin treated patients (30.8 and 32.1%, respectively). There was a consistent significant reduction of hypoglycemia risk associated with insulin glargine, compared with NPH insulin, in terms of overall symptomatic (11%; P = 0.0006) and nocturnal (26%; P < 0.0001) hypoglycemia. Most notably, the risk of severe hypoglycemia and severe nocturnal hypoglycemia were reduced with insulin glargine by 46% (P = 0.0442) and 59% (P = 0.0231), respectively. CONCLUSIONS: These results confirmed that insulin glargine given once daily reduces the risk of hypoglycemia compared with NPH insulin, which can facilitate more aggressive insulin treatment to a HbA(1c) target of < or =7.0% in patients with type 2 diabetes. PMID- 15793206 TI - Diabetic neuropathies: a statement by the American Diabetes Association. PMID- 15793207 TI - Diabetic retinopathy and neuropathy. PMID- 15793208 TI - Primary prevention of cardiovascular disease in pre-diabetes: the glass is half full and half empty. PMID- 15793210 TI - Counterpoint: Inpatient glucose management: a premature call to arms? PMID- 15793209 TI - Point: Inpatient glucose management: the emperor finally has clothes. PMID- 15793211 TI - Spontaneous diabetic myonecrosis. PMID- 15793212 TI - The role of platelet-activating factor in pathogenesis of type 1 diabetes. PMID- 15793213 TI - Potentially important contribution of dextrose used as diluent to hyperglycemia in hospitalized patients. PMID- 15793214 TI - Use of insulin glargine during the first weeks of pregnancy in five type 1 diabetic women. PMID- 15793215 TI - Successful treatment of insulin allergy in a 1-year-old infant with neonatal diabetes by lispro and glargine insulin. PMID- 15793216 TI - Charcot neuroarthropathy of the wrist in type 1 diabetes. PMID- 15793217 TI - Lipids and glucose in type 2 diabetes: what about the beta-cell and the mitochondria?: response to Boden and Laakso. PMID- 15793219 TI - Adjuvant post-operative chemoradiotherapy in head and neck cancer: a standard of care? PMID- 15793220 TI - FDA drug approval summary: azacitidine (5-azacytidine, Vidaza) for injectable suspension. AB - On May 19, 2004, azacitidine (5-azacytidine; Vidaza(trade mark); Pharmion Corporation, Boulder, CO, http://www.pharmion.com) for injectable suspension received regular approval by the U.S. Food and Drug Administration (FDA) for the treatment of all subtypes of myelodysplastic syndrome (MDS). This report summarizes the basis for this approval. Effectiveness was demonstrated in one randomized, controlled trial comparing azacitidine administered s.c. with best supportive care (observation group) and in two single-arm studies, one in which azacitidine was administered s.c. and in the other in which it was administered i.v. The dose of azacitidine, 75 mg/m2/day for 7 days every 28 days, was the same in all three studies. In the randomized trial, study participants were well matched with respect to age, sex, race, performance status, MDS subtype, and use of transfusion during the 3 months before study entry. Patients in the observation arm were permitted by protocol to cross over to azacitidine treatment if their disease progressed according to prespecified criteria. During the course of the study, more than half of the patients in the observation arm did cross over to the azacitidine treatment arm. The primary efficacy end point was the overall response rate. Response consisted of complete or partial normalization of blood cell counts and of bone marrow morphology. The response rate in the azacitidine arm was about 16%; there were no responses in the observation arm. The response rates in the two single-arm studies were similar (13% and 19%). The responses were sustained, with median durations of 11 months and 17 months respectively. Responding patients who were transfusion dependent at study entry lost the need for transfusions. In addition, about 19% of patients had less than partial responses (termed improvement), and two-thirds of them became transfusion independent. Common adverse events associated with azacitidine treatment were gastrointestinal (nausea, vomiting, diarrhea, constipation, and anorexia), hematologic (neutropenia, thrombocytopenia), fevers, rigors, ecchymoses, petechiae, injection site events, arthralgia, headache, and dizziness. Liver function abnormalities occurred in 16% of patients with intercurrent hepatobiliary disorders and in two patients with previously diagnosed liver cirrhosis. Renal failure occurred in patients during sepsis and hypotension. There were no deaths attributed to azacitidine. Azacitidine, the first drug approved by the U.S. FDA for MDS, has a favorable safety profile and provides a clinical benefit of eliminating transfusion dependence and complete or partial normalization of blood counts and bone marrow blast percentages in responding patients. PMID- 15793221 TI - Phase II trial of oral rubitecan in previously treated pancreatic cancer patients. AB - BACKGROUND: Additional systemic treatments for locally advanced or metastatic pancreatic cancer are needed, as current treatment options produce only modest survival benefits. Rubitecan (Orathecin; Supergen Inc., Dublin, CA, http://www.supergen.com) is an orally active camptothecin derivative with demonstrated responses in patients with pancreatic cancer in early clinical trials. This phase II, open-label trial was developed to assess the safety and efficacy of rubitecan in patients with locally advanced or metastatic pancreatic cancer refractory to conventional chemotherapy. METHODS: Fifty-eight patients with failed or relapsed advanced pancreatic cancer after receiving at least one prior chemotherapy regimen were enrolled to receive eight consecutive weeks of treatment with rubitecan at a dose of 1.5 mg/m2 orally on five consecutive days per week, followed by 2 days off therapy, repeatedly. The primary end point was response rate. Time to progression, overall survival, changes in CA19-9 levels, and the composite measure of clinical benefit response were evaluated as secondary end points. RESULTS: Among 43 patients with measurable disease, 7% (3/43) achieved partial responses and 16% (7/43) had disease stabilization for an overall response and disease stabilization rate of 23%. All responses were confirmed by independent radiology review. Median survival was longer in responding patients than in the overall study cohort (10 months versus 3 months). Gastrointestinal and hematologic toxicities were the most commonly reported adverse events. CONCLUSION: Oral rubitecan produced responses and was well tolerated by heavily pretreated patients with refractory pancreatic cancer. The overall risk-benefit profile of oral rubitecan appears promising, supporting further evaluation in phase III trials in patients with refractory and chemotherapy-naive pancreatic cancer. PMID- 15793222 TI - VEGF-targeted therapy in metastatic renal cell carcinoma. AB - PURPOSE: To review the biology of renal cell carcinoma (RCC) and the clinical results of vascular endothelial growth factor (VEGF) blockade in metastatic RCC. METHODS: A review of relevant published literature regarding VEGF, von Hippel Lindau (VHL) gene inactivation, and VEGF overexpression in RCC was performed. Further, a review of the mechanism, toxicity, and clinical development of VEGF targeted therapy in metastatic RCC was undertaken. RESULTS: VHL tumor suppressor gene inactivation is observed in the majority of clear cell RCC cases, leading to VEGF overexpression. Therapy with agents directed against the VEGF protein or the VEGF receptor have demonstrated initial clinical activity in metastatic RCC. CONCLUSIONS: Therapeutic targeting of VEGF in RCC has strong biologic rationale. Substantial clinical activity has been reported in initial clinical trials with VEGF-targeting agents. Further investigation is needed to optimally use these agents for maximal clinical benefit. PMID- 15793223 TI - Never too old? Age should not be a barrier to enrollment in cancer clinical trials. AB - Throughout Europe and the U.S., over 60% of the total incidence of cancer occurs in the elderly (> or =65 years) population, a patient group that requires particular consideration when making treatment decisions due to a number of factors. Despite this, elderly patients are generally under-represented in clinical trials such that study data should be interpreted with caution because results in younger cancer patients may not always extrapolate to the typical elderly cancer patient. Reports suggest that elderly cancer patients represent around 22% of patients enrolled in phase II clinical studies. Barriers to the accrual of elderly patients to clinical trials include lack of appropriate trials, high burden of comorbidity, study-imposed restrictions, and attitudes of physicians. There is a belief that elderly patients may be unable to tolerate various cancer therapies, which may result in this patient population being excluded from prospective trials. However, clinical data demonstrate that age alone is not a sufficient reason to withhold treatment. Lack of clinical trial data and the associated lack of evidence-based guidelines for elderly patients mean physicians have little to guide them, with the result that patients may not receive the optimal therapy. As clinical trials are the primary method of evaluating the efficacy and safety of adjuvant and palliative cancer therapies, trials that specifically target the elderly cancer patient are required to adequately assess the risks and benefits of treatment in this vulnerable population. This review aims to assess the clinical reality and clinical trial age mismatch to evaluate implications for elderly cancer patients and to identify how this situation may be addressed. Possible reasons for the disparity, and the resulting clinical consequences, are also considered. PMID- 15793224 TI - Pegylated liposomal doxorubicin: optimizing the dosing schedule in ovarian cancer. AB - The need for effective, well-tolerated, and convenient therapies for patients with advanced ovarian cancer has led researchers to continually refine chemotherapeutic regimens to balance efficacy with safety and tolerability in order to maintain or improve patient quality of life. In this article, we review current strategies for the optimal dosing of pegylated liposomal doxorubicin (DOXIL; Tibotec Therapeutics, a division of Ortho Biotech Products, L.P., Bridgewater, NJ, http://www.tibotec.com; Caelyx, Schering-Plough Corporation, Kenilworth, NJ, http://www.sch-plough.com) in relapsed ovarian cancer. Pegylated liposomal doxorubicin has demonstrated efficacy in the treatment of recurrent/resistant ovarian cancer in several clinical trials utilizing a dose of 50 mg/m2 every 4 weeks. The most common adverse events associated with pegylated liposomal doxorubicin treatment in these studies-hand-foot syndrome (HFS, also known as palmar-plantar erythrodysesthesia) and stomatitis-are schedule and dose dependent, respectively, and do not typically lead to discontinuation of therapy. Several phase II and retrospective studies support the use of pegylated liposomal doxorubicin 40 mg/m2 every 4 weeks (dose intensity of 10 mg/m2 weekly) to optimize clinical efficacy and minimize the occurrence of schedule- and dose related adverse events in patients with recurrent/relapsed ovarian cancer. Further reductions in dose intensity are necessary for use in combined chemotherapy regimens. Antitumor activity was maintained, with reduced incidences of HFS and stomatitis. Given the chronic course of ovarian cancer, the improved tolerability profile of pegylated liposomal doxorubicin 40 mg/m2 combined with a convenient once-monthly dosing schedule may translate into an improved quality of life for patients with ovarian cancer. PMID- 15793225 TI - Chemoradiation after surgery for high-risk head and neck cancer patients: how strong is the evidence? AB - Patients with locally advanced, operable head and neck squamous cell carcinoma (HNSCC) are known to be at high risk of treatment failure, ranging from local regrowth to lymphatic spread to systemic dissemination. Attacking specifically each of these patterns of failure implies the use of a multimodal approach. Throughout the past two decades the management of stages III/IV HNSCC remained a matter of debate, especially with regards to treatment intensity and sequencing. Surgery and/or radiotherapy were the mainstay of local-regional treatment in patients with locally advanced disease, but treatment outcome often remained disappointing. In the hope of improving the prognosis after radical surgery, cisplatin-based combinations have been administered before surgery, in the interval between surgery and radiotherapy, or after radiotherapy. Until very recently these combinations, at best, decreased systemic failures without having a real impact on local outcome or survival. Indeed, until the mid-1990s, most trials that had tested postoperative combinations of chemotherapy and radiotherapy did not show any significant benefit. In 2004 level I evidence was established with the publication of the results of two large-scale, independent but similar trials conducted in Europe and the U.S. Both studies demonstrated that, compared with postoperative irradiation alone, adjuvant concurrent chemoradiation was more efficacious in terms of local-regional control and disease-free survival. With the publication of these two trials the evidence demonstrating the potential value of concurrent postoperative chemoradiotherapy in high-risk operable head and neck cancer is strong; however, additional studies and comparative analysis of the selection criteria and treatment outcomes across these two trials will be needed to gain a more accurate assessment of benefit and risk levels in specific patients with operable, locally advanced disease. PMID- 15793226 TI - Increased hemoglobin levels and improved quality-of-life assessments during epoetin alfa treatment in anemic cancer patients: results of a prospective, multicenter German trial. AB - This prospective, open-label, multicenter study was undertaken to determine the safety and efficacy of epoetin alfa in increasing hemoglobin levels and improving quality of life (QOL), specifically fatigue, in cancer patients receiving chemotherapy with or without radiotherapy (n=702). Epoetin alfa, 10,000 IU three times a week s.c. for 8-18 weeks, increased the mean hemoglobin level relative to baseline (1.0 +/- 1.5 g/dl by week 4 and > or =1.7 g/dl from week 10 through the end of the trial), with 63.4% of patients experiencing > or =2 g/dl increases in hemoglobin above baseline at some time during the study. Fatigue is an important component of QOL. Physicians, nurses, and patients independently assessed patient fatigue level on a linear-analogue scale. Although all three groups reported improvements in patient fatigue over the course of the study (p <.0001), the magnitude of fatigue ratings and their relationship to tumor response and to hemoglobin level varied by group. Overall, epoetin alfa was well tolerated and effective in improving hemoglobin levels and decreasing fatigue in patients undergoing chemotherapy. PMID- 15793227 TI - The molecular perspective: L-asparaginase. PMID- 15793228 TI - A burning question: does an adipokine-induced activation of the immune system mediate the effect of overnutrition on type 2 diabetes? AB - There is growing support for the hypothesis that obesity is an inflammatory condition leading to chronic activation of the innate immune system, which ultimately causes progressive impairment of glucose tolerance. Experimental studies in animals and evidence from prospective and longitudinal studies in humans are consistent with an etiologic role of subclinical inflammation in the pathogenesis of type 2 diabetes, primarily as a mediator of obesity-induced insulin resistance. However, the exact chain of molecular events linking overnutrition, activation of the innate immune system, and impairment of insulin signaling in peripheral tissues remains incompletely understood. Notwithstanding this limitation, treating the underlying subclinical inflammation may constitute a novel approach to prevention and/or treatment of type 2 diabetes. PMID- 15793229 TI - Long-term AICAR administration and exercise prevents diabetes in ZDF rats. AB - Lifestyle interventions including exercise programs are cornerstones in the prevention of obesity-related diabetes. The AMP-activated protein kinase (AMPK) has been proposed to be responsible for many of the beneficial effects of exercise on glucose and lipid metabolism. The effects of long-term exercise training or 5-aminoimidazole-4-carboxamide-1-beta-d-riboruranoside (AICAR) treatment, both known AMPK activators, on the development of diabetes in male Zucker diabetic fatty (ZDF) rats were examined. Five-week-old, pre-diabetic ZDF rats underwent daily treadmill running or AICAR treatment over an 8-week period and were compared with an untreated group. In contrast to the untreated, both the exercised and AICAR-treated rats did not develop hyperglycemia during the intervention period. Whole-body insulin sensitivity, as assessed by a hyperinsulinemic-euglycemic clamp at the end of the intervention period, was markedly increased in the exercised and AICAR-treated animals compared with the untreated ZDF rats (P < 0.01). In addition, pancreatic beta-cell morphology was almost normal in the exercised and AICAR-treated animals, indicating that chronic AMPK activation in vivo might preserve beta-cell function. Our results suggest that activation of AMPK may represent a therapeutic approach to improve insulin action and prevent a decrease in beta-cell function associated with type 2 diabetes. PMID- 15793230 TI - Altered hepatic and muscle substrate utilization provoked by GLUT4 ablation. AB - Studies were conducted to explore altered substrate utilization and metabolism in GLUT4 null mice. Liver fatty acid synthase mRNA and fatty acid synthesis rates were dramatically increased in GLUT4 null mice compared with control mice and were supported by increased rates of the pentose phosphate pathway oxidative phase and sterol regulatory binding protein mRNA expression. Increased GLUT2 protein content, glucokinase mRNA, and glucose-6-phosphate in GLUT4 null mice may provide substrate for the enhanced fatty acid synthesis. Increased fatty acid synthesis, however, did not lead to hepatic triglyceride accumulation in GLUT4 null mice because of increased hepatic triglyceride secretion rates. GLUT4 null mice rapidly cleared orally administered olive oil, had reduced serum triglyceride concentrations in the fed and the fasted state, and increased skeletal muscle lipoprotein lipase when compared with controls. Oleate oxidation rates were increased in GLUT4 null skeletal muscle in association with mitochondrial hyperplasia/hypertrophy. This study demonstrated that GLUT4 null mice had increased hepatic glucose uptake and conversion into triglyceride for subsequent use by muscle. The ability of GLUT4 null mice to alter hepatic carbohydrate and lipid metabolism to provide proper nutrients for peripheral tissues may explain (in part) their ability to resist diabetes when fed a normal diet. PMID- 15793231 TI - Modular kinetic analysis of the adenine nucleotide translocator-mediated effects of palmitoyl-CoA on the oxidative phosphorylation in isolated rat liver mitochondria. AB - To test whether long-chain fatty acyl-CoA esters link obesity with type 2 diabetes through inhibition of the mitochondrial adenine nucleotide translocator, we applied a system-biology approach, dual modular kinetic analysis, with mitochondrial membrane potential (Deltapsi) and the fraction of matrix ATP as intermediates. We found that 5 mumol/l palmitoyl-CoA inhibited adenine nucleotide translocator, without direct effect on other components of oxidative phosphorylation. Indirect effects depended on how oxidative phosphorylation was regulated. When the electron donor and phosphate acceptor were in excess, and the mitochondrial "work" flux was allowed to vary, palmitoyl-CoA decreased phosphorylation flux by 38% and the fraction of ATP in the medium by 39%. Deltapsi increased by 15 mV, and the fraction of matrix ATP increased by 46%. Palmitoyl-CoA had a stronger effect when the flux through the mitochondrial electron transfer chain was maintained constant: Deltapsi increased by 27 mV, and the fraction of matrix ATP increased 2.6 times. When oxidative phosphorylation flux was kept constant by adjusting the rate using hexokinase, Deltapsi and the fraction of ATP were not affected. Palmitoyl-CoA increased the extramitochondrial AMP concentration significantly. The effects of palmitoyl-CoA in our model system support the proposed mechanism linking obesity and type 2 diabetes through an effect on adenine nucleotide translocator. PMID- 15793232 TI - Acute induction of gene expression in brain and liver by insulin-induced hypoglycemia. AB - The robust neuroendocrine counterregulatory responses induced by hypoglycemia protect the brain by restoring plasma glucose, but little is known about molecular responses to hypoglycemia that may also be neuroprotective. To clarify these mechanisms, we examined gene expression in hypothalamus, cortex, and liver 3 h after induction of mild hypoglycemia by a single injection of insulin, using cDNA microarray analysis and quantitative real-time PCR. Real-time PCR corroborated the induction of six genes (angiotensinogen, GLUT-1, inhibitor of kappaB, inhibitor of DNA binding 1 [ID-1], Ubp41, and mitogen-activated protein kinase phosphatase-1 [MKP-1]) by insulin-induced hypoglycemia in the hypothalamus: five of these six genes in cortex and three (GLUT-1, angiotensinogen, and MKP-1) in liver. The induction was due to hypoglycemia and not hyperinsulinemia, since fasting (characterized by low insulin and glucose) also induced these genes. Four of these genes (angiotensinogen, GLUT-1, ID-1, and MKP-1) have been implicated in enhancement of glucose availability, which could plausibly serve a neuroprotective role during acute hypoglycemia but, if persistent, could also cause glucose-sensing mechanisms to overestimate plasma glucose levels, potentially causing hypoglycemia-induced counterregulatory failure. Although using cDNA microarrays with more genes, or microdissection, would presumably reveal further responses to hypoglycemia, these hypoglycemia induced genes represent useful markers to assess molecular mechanisms mediating cellular responses to hypoglycemia. PMID- 15793233 TI - S-nitrosation of the insulin receptor, insulin receptor substrate 1, and protein kinase B/Akt: a novel mechanism of insulin resistance. AB - Evidence demonstrates that exogenous nitric oxide (NO) and the NO produced by inducible nitric oxide synthase (iNOS) can induce insulin resistance in muscle. Here, we investigated whether this insulin resistance could be mediated by S nitrosation of proteins involved in early steps of the insulin signal transduction pathway. Exogenous NO donated by S-nitrosoglutathione (GSNO) induced in vitro and in vivo S-nitrosation of the insulin receptor beta subunit (IRbeta) and protein kinase B/Akt (Akt) and reduced their kinase activity in muscle. Insulin receptor substrate (IRS)-1 was also rapidly S-nitrosated, and its expression was reduced after chronic GSNO treatment. In two distinct models of insulin resistance associated with enhanced iNOS expression-diet-induced obesity and the ob/ob diabetic mice-we observed enhanced S-nitrosation of IRbeta/IRS-1 and Akt in muscle. Reversal of S-nitrosation of these proteins by reducing iNOS expression yielded an improvement in insulin action in both animal models. Thus, S-nitrosation of proteins involved in insulin signal transduction is a novel molecular mechanism of iNOS-induced insulin resistance. PMID- 15793235 TI - Chronic hyperglycemia enhances PEPCK gene expression and hepatocellular glucose production via elevated liver activating protein/liver inhibitory protein ratio. AB - Acute hyperglycemia normally suppresses hepatic glucose production (HGP) and gluconeogenic gene expression. Conversely, chronic hyperglycemia is accompanied by progressive increases in basal HGP and is a major contributor to hyperglycemia in both type 1 and type 2 diabetes by mechanisms that are poorly understood. The aim of this study was to investigate the molecular mechanisms whereby hyperglycemia contributes to excessive gluconeogenesis in Fao hepatoma cells. Increasing glucose from 5 to 20 mmol/l resulted in loss of glucose inhibition of PEPCK gene expression after 12 h. Furthermore, 24 h of incubation with 20 mmol/l glucose increased cAMP-stimulated PEPCK mRNA by approximately 40% (P < 0.05) and similarly increased glucose production. Although total CCAAT/enhancer-binding protein beta (C/EBPbeta) protein levels were suppressed, 20 mmol/l glucose increased the liver activating protein (LAP; an active isoform of C/EBPbeta)/liver inhibitory protein (LIP; an inhibitory isoform of C/EBPbeta) ratio significantly. Chromatin immunoprecipitation studies of the endogenous PEPCK gene demonstrated an increased association of LAP with the cAMP response element of the promoter. Using transient transfection to manipulate the LAP/LIP ratio, we also demonstrate a direct relationship between this ratio and PEPCK promoter activity. An increased LAP/LIP ratio not only enhanced cAMP- and dexamethasone-induced PEPCK gene expression but also impaired the repressive effect of insulin. These results demonstrate that sustained hyperglycemia diminishes the inhibitory effect of glucose and insulin on PEPCK expression and enhances hormone-stimulated PEPCK gene expression and hepatocellular glucose production. Because prolonged hyperglycemia increases the LAP/LIP ratio and can potentiate hormone induction of PEPCK transcription, our results suggest that a hyperglycemia-driven increased LAP/LIP ratio may be a critical molecular event in the pathogenesis of increased HGP in diabetes. PMID- 15793234 TI - Endoplasmic reticulum stress-induced apoptosis is partly mediated by reduced insulin signaling through phosphatidylinositol 3-kinase/Akt and increased glycogen synthase kinase-3beta in mouse insulinoma cells. AB - An imbalance between the rate of protein synthesis and folding capacity of the endoplasmic reticulum (ER) results in stress that has been increasingly implicated in pancreatic islet beta-cell apoptosis and diabetes. Because insulin/IGF/Akt signaling has been implicated in beta-cell survival, we sought to determine whether this pathway is involved in ER stress-induced apoptosis. Mouse insulinoma cells treated with pharmacological agents commonly used to induce ER stress exhibited apoptosis within 48 h. ER stress-induced apoptosis was inhibited by cotreatment of the cells with IGF-1. Stable cell lines were created by small interfering RNA (siRNA) with graded reduction of insulin receptor expression, and these cells had enhanced susceptibility to ER stress-induced apoptosis and reduced levels of phospho-glycogen synthase kinase 3beta (GSK3beta). In control cells, ER stress-induced apoptosis was associated with a reduction in phospho-Akt and phospho-GSK3beta. To further assess the role of GSK3beta in ER stress-induced apoptosis, stable cell lines were created by siRNA with up to 80% reduction in GSK3beta expression. These cells were found to resist ER stress-induced apoptosis. These results illustrate that ER stress-induced apoptosis is mediated at least in part by signaling through the phosphatidylinositol 3 kinase/Akt/GSK3beta pathway and that GSK3beta represents a novel target for agents to promote beta-cell survival. PMID- 15793236 TI - Insulin, C-peptide, and leptin concentrations predict increased visceral adiposity at 5- and 10-year follow-ups in nondiabetic Japanese Americans. AB - We prospectively examined the relationship between leptin and markers of insulin resistance and secretion and future visceral adipose tissue accumulation. In this study, 518 nondiabetic Japanese-American men and women underwent the following measurements at baseline and at 5- and 10-year follow-ups: plasma glucose and insulin measured after an overnight fast and during a 75-g oral glucose tolerance test, insulin secretion ratio (ISR) [(30-min insulin - fasting insulin)/30-min glucose], fasting C-peptide levels, plasma leptin (baseline only), and fat areas (intra-abdominal and subcutaneous) measured by computed tomography. Predictors of future intra-abdominal fat (IAF) were determined using multiple linear regression. Fasting insulin and C-peptide levels at baseline were significantly associated with IAF area at 5 years (coefficient = 0.041, P = 0.001 and coefficient = 1.283, P < 0.001, respectively) and 10 years (coefficient = 0.031, P = 0.020 and coefficient = 0.221, P = 0.035, respectively). ISR was not significantly associated with IAF at 5 or 10 years. Leptin level at baseline was positively associated with IAF at 5 years (coefficient = 0.055, P = 0.002) and 10 years (coefficient = 0.059, P = 0.003). In conclusion, higher levels of fasting insulin, C-peptide, and circulating leptin level predicted visceral fat accumulation independent from subcutaneous fat accumulation in nondiabetic Japanese-American men and women in both short-term (5 years) and long-term (10 years) follow-up. PMID- 15793237 TI - Deletion of the angiotensin type 2 receptor (AT2R) reduces adipose cell size and protects from diet-induced obesity and insulin resistance. AB - The renin-angiotensin system with its active metabolite angiotensin (Ang) II has been related not only to hypertension but also to obesity and insulin resistance. Recent evidence obtained in vitro suggests that the type 2 Ang II receptor (AT2R) mediates the trophic action of Ang II on adipocyte differentiation and lipogenesis. We used AT2R(y/-) mice to delineate a potential role of AT2R in adipose tissue development and metabolism. AT2R(y/-) mice had a normal adiposity but displayed a striking adipose tissue phenotype characterized by small adipocytes and an increase in cell number. In muscle, the expression of several genes involved in lipid metabolism, including fatty acid translocase, uncoupling protein-3, peroxisome proliferator-activated receptors (alpha, delta), and carnitine palmitoyl transferase-1, was increased in AT2R-deficient mice. In response to high-fat feeding, these mice were protected against obesity and obesity-related glucose intolerance, as assessed by glucose tolerance tests. Moreover, lipid oxidation assessed by indirect calorimetry was higher in AT2R deficient mice than in wild-type mice, irrespective of the diet. This suggests that AT2R-dependent signaling exerts a direct or indirect negative control on lipid utilization in muscles. These data support the idea that AT2R-dependent Ang II signaling increases adipose cell mass and glucose intolerance and thus could participate to the deleterious effects of a high-fat diet. PMID- 15793238 TI - Androgen receptor null male mice develop late-onset obesity caused by decreased energy expenditure and lipolytic activity but show normal insulin sensitivity with high adiponectin secretion. AB - Androgen receptor (AR) null male mice (AR(L-/Y)) revealed late-onset obesity, which was confirmed by computed tomography-based body composition analysis. AR(L /Y) mice were euphagic compared with the wild-type male (AR(X/Y)) controls, but they were also less dynamic and consumed less oxygen. Transcript profiling indicated that AR(L-/Y) mice had lower transcripts for the thermogenetic uncoupling protein 1, which was subsequently found to be ligand-dependently activated by AR. We also found enhanced secretion of adiponectin, which is insulin sensitizing, from adipose tissue and a relatively lower expression of peroxisome proliferator-activated receptor-gamma in white adipose tissue in comparison to AR(X/Y) mice. Both factors might explain why the overall insulin sensitivity of AR(L-/Y) mice remained intact, despite their apparent obesity. The results revealed that AR plays important roles in male metabolism by affecting the energy balance, and it is negative to both adiposity and insulin sensitivity. PMID- 15793239 TI - PDX-1/VP16 fusion protein, together with NeuroD or Ngn3, markedly induces insulin gene transcription and ameliorates glucose tolerance. AB - Diabetes is the most prevalent and serious metabolic disease, and the number of diabetic patients worldwide is increasing. The reduction of insulin biosynthesis in pancreatic beta-cells is closely associated with the onset and progression of diabetes, and thus it is important to search for ways to induce insulin-producing cells in non-beta-cells. In this study, we showed that a modified form of the pancreatic and duodenal homeobox factor 1 (PDX-1) carrying the VP16 transcriptional activation domain (PDX-1/VP16) markedly increases insulin biosynthesis and induces various pancreas-related factors in the liver, especially in the presence of NeuroD or neurogenin 3 (Ngn3). Furthermore, in streptozotocin-induced diabetic mice, PDX-1/VP16 overexpression, together with NeuroD or Ngn3, drastically ameliorated glucose tolerance. Thus PDX-1/VP16 expression, together with NeuroD or Ngn3, markedly induces insulin gene transcription and ameliorates glucose tolerance. This approach warrants further investigation and may have utility in the treatment of diabetes. PMID- 15793241 TI - Reversal of diabetes in pancreatectomized pigs after transplantation of neonatal porcine islets. AB - Neonatal porcine islets (NPIs) are able to grow and to reverse hyperglycemia after transplantation in immunoincompetent mice. The aim of this study was to demonstrate the feasibility of allogeneic NPI grafts to achieve normoglycemia in a pancreatectomized diabetic pig. NPIs were isolated from pancreases of 1- to 3 day-old pigs, cultured, and then transplanted via the portal vein into the liver of totally pancreatectomized pigs (mean body weight, 20.8 kg). Each pig received NPIs consisting of 3.1 +/- 0.3 x 10(6) beta-cells/kg (12,476 +/- 1,146 islet equivalent/kg). The six pigs that were given cyclosporine and sirolimus achieved normoglycemia by day 14 without insulin therapy. Three pigs died of surgical complications shortly after transplantation, whereas the other three remained insulin independent up to day 69. Of seven nonimmunosuppressed recipients, four pigs became normoglycemic by day 14 without insulin treatment, with two of the animals remaining normoglycemic long term. Well-preserved insulin-positive cells were found in the graft at the end of follow-up with a significant increase in insulin content in long-term survivors of both groups. This study demonstrates for the first time that allogeneic NPIs can reverse hyperglycemia in totally pancreatectomized diabetic pigs. PMID- 15793240 TI - Adipocyte-specific glucocorticoid inactivation protects against diet-induced obesity. AB - Local glucocorticoid (GC) action depends on intracellular GC metabolism by 11beta hydroxysteroid dehydrogenases (11betaHSDs). 11betaHSD1 activates GCs, while 11betaHSD2 inactivates GCs. Adipocyte-specific amplification of GCs through transgenic overexpression of 11betaHSD1 produces visceral obesity and the metabolic syndrome in mice. To determine whether adipocyte-specific inactivation of GCs protects against this phenotype, we created a transgenic model in which human 11betaHSD2 is expressed under the control of the murine adipocyte fatty acid binding protein (aP2) promoter (aP2-h11betaHSD2). Transgenic mice have increased 11betaHSD2 expression and activity exclusively in adipose tissue, with the highest levels in subcutaneous adipose tissue, while systemic indexes of GC exposure are unchanged. Transgenic mice resist weight gain on high-fat diet due to reduced fat mass accumulation. This improved energy balance is associated with decreased food intake, increased energy expenditure, and improved glucose tolerance and insulin sensitivity. Adipose tissue gene expression in transgenic mice is characterized by decreased expression of leptin and resistin and increased expression of adiponectin, peroxisome proliferator-activated receptor gamma, and uncoupling protein 2. These data suggest that reduction of active GCs exclusively in adipose tissue is an important determinant of a favorable metabolic phenotype with respect to energy homeostasis and the metabolic syndrome. PMID- 15793242 TI - In vivo control of diabetogenic T-cells by regulatory CD4+CD25+ T-cells expressing Foxp3. AB - To understand the ability of regulatory T-cells to control diabetes development in clinically relevant situations, we established a new model of accelerated diabetes in young DP-BB rats by transferring purified T-cells from DR-BB rats made acutely diabetic. Transfer of 3, 5, 10, or 23 million pure in vitro activated T-cells accelerated diabetes onset in >90% of the recipients, with the degree of acceleration being dosage dependent. Cotransfer of unfractionated leukocytes from healthy donors prevented diabetes. Full protection was achieved when protective cells were transferred 3-4 days before diabetogenic cells, whereas transfer 2 days before conferred only partial protection. Protection resided in the CD4(+) fraction, as purified CD4(+) T-cells prevented the accelerated diabetes. When CD25(+) cells were depleted from these cells before they were transferred, their ability to prevent diabetes was impaired. In contrast, two million CD4(+)CD25(+) cells (expressing Foxp3) prevented the accelerated diabetes when transferred both before and simultaneously with the diabetogenic T-cells. In addition, 2 million CD4(+)CD25(+) T-cells prevented spontaneous diabetes, even when given to rats age 42 days, whereas 20 million CD4(+)CD25(-) cells (with low Foxp3 expression) were far less effective. We thus demonstrated that CD4(+)CD25(+) cells exhibit powerful regulatory potential in rat diabetes. PMID- 15793243 TI - Achieving permanent survival of islet xenografts by independent manipulation of direct and indirect T-cell responses. AB - Recent success in pancreatic islet allotransplantation has raised expectations but has equally highlighted the acute shortage of donor tissue. The use of xenogeneic tissue would help to address this shortage; however, strong cellular immunity limits the application of this approach. T-cell responses to xenogeneic tissues involve recognition of intact species-mismatched major histocompatibility complex (MHC) molecules, the direct pathway, and xenogeneic proteins presented as peptides by responder-type MHC molecules, the indirect pathway. In this study, we exploited the species difference to selectively and sequentially inhibit direct and indirect xenoresponses after transplantation of porcine islets into mice. Selective inhibition of the direct response was achieved using porcine CTLA4-Ig, which binds preferentially to pig versus mouse B7 molecules. Selective inhibition of the indirect response was achieved using murine CTLA4-Ig, which binds preferentially to mouse B7 molecules. Administration of porcine CTLA4-Ig alone caused modest prolongation of islet survival. Injection of murine CTLA4-Ig alone had a minimal effect. However, the injection of the porcine fusion protein early and the murine homolog late after grafting led to permanent survival of the porcine islets, in the absence of any other immunosuppression. These results suggest that a similar approach could have clinical utility in porcine islet xenotransplantation. PMID- 15793244 TI - Distinct effects of glucose-dependent insulinotropic polypeptide and glucagon like peptide-1 on insulin secretion and gut motility. AB - Glucose-induced insulin secretion from pancreatic beta-cells depends critically on ATP-sensitive K(+) channel (K(ATP) channel) activity, but it is not known whether K(ATP) channels are involved in the potentiation of insulin secretion by glucose-dependent insulinotropic polypeptide (GIP). In mice lacking K(ATP) channels (Kir6.2(-/-) mice), we found that pretreatment with GIP in vivo failed to blunt the rise in blood glucose levels after oral glucose load. In Kir6.2(-/-) mice, potentiation of insulin secretion by GIP in vivo was markedly attenuated, indicating that K(ATP) channels are essential in the insulinotropic effect of GIP. In contrast, pretreatment with glucagon-like peptide-1 (GLP-1) in Kir6.2(-/ ) mice potentiated insulin secretion and blunted the rise in blood glucose levels. We also found that GLP-1 inhibited gut motility whereas GIP did not. Perfusion experiments of Kir6.2(-/-) mice revealed severely impaired potentiation of insulin secretion by 1 nmol/l GIP and substantial potentiation by 1 nmol/l GLP 1. Although both GIP and GLP-1 increase the intracellular cAMP concentration and potentiate insulin secretion, these results demonstrate that the GLP-1 and GIP signaling pathways involve the K(ATP) channel differently. PMID- 15793247 TI - Inducible nitric oxide synthase has divergent effects on vascular and metabolic function in obesity. AB - Previous studies have suggested an involvement of inducible nitric oxide synthase (iNOS) in obesity, but the relation, if any, between this and mechanisms underlying endothelial dysfunction in obesity is unknown. We studied mice fed an obesogenic high-fat or standard diet for up to 8 weeks. Obesity was associated with elevated blood pressure; resistance to the glucoregulatory actions of insulin; resistance to the vascular actions of insulin, assessed as the reduction in phenylephrine constrictor response of aortic rings after insulin preincubation (lean -21.7 +/- 11.5 vs. obese 18.2 +/- 15.5%; P < 0.05); and evidence of reactive oxygen species (ROS)-dependent vasodilatation in response to acetylcholine in aortic rings (change in maximal relaxation to acetylcholine after exposure to catalase: lean -2.1 +/- 6.0 vs. obese -15.0 +/- 3.8%; P = 0.04). Obese mice had increased expression of iNOS in aorta, with evidence of increased vascular NO production, assessed as the increase in maximal constriction to phenylephrine after iNOS inhibition with 1400W (lean -3.5 +/- 9.1 vs. obese 42.1 +/- 11.2%; P < 0.001). To further address the role of iNOS in obesity-induced vascular and metabolic dysfunction, we studied the effect of a high-fat diet in iNOS knockout mice (iNOS KO). Obese iNOS KO mice were protected against the development of resistance to insulin's glucoregulatory and vascular effects (insulin-dependent reduction in maximal phenylephrine response: obese wild-type 11.2 +/- 15.0 vs. obese iNOS KO -20.0 +/- 7.7%; P = 0.02). However, obese iNOS KO mice remained hypertensive (124.0 +/- 0.7 vs. 114.9 +/- 0.5 mmHg; P < 0.01) and had evidence of increased vascular ROS production. Although these data support iNOS as a target to protect against the adverse effects of obesity on glucoregulation and vascular insulin resistance, iNOS inhibition does not prevent the development of raised blood pressure or oxidative stress. PMID- 15793246 TI - Pancreatic beta-cell failure and diabetes in mice with a deletion mutation of the endoplasmic reticulum molecular chaperone gene P58IPK. AB - The endoplasmic reticulum (ER) transmits apoptotic signals in the pancreas during ER stress, implicating ER stress-mediated apoptosis in the development of diabetes. P58(IPK) (DNAJC3) is induced during ER stress and functions as a negative feedback component to inhibit eIF-2alpha signaling and attenuate the later phases of the ER stress response. To gain insight into a more comprehensive role of P58(IPK) function, we generated deletion mutant mice that showed a gradual onset of glucosuria and hyperglycemia associated with increasing apoptosis of pancreatic islet cells. Lack of P58(IPK) had no apparent effect on the functional integrity of viable beta-cells. A set of genes associated with apoptosis showed altered expression in pancreatic islets from P58(IPK)-null mice, further substantiating the apoptosis phenotype. The data provide in vivo evidence to support the concept that P58(IPK) functions as a signal for the downregulation of ER-associated proteins involved in the initial ER stress response, thus preventing excessive cell loss by degradation pathways. Insulin deficiency associated with the absence of P58(IPK) mimics beta-cell failure associated with type 1 and late-stage type 2 diabetes. P58(IPK) function and activity may therefore provide a novel area of investigation into ER-mediated mechanistic and therapeutic approaches for diabetes. PMID- 15793245 TI - Neuronatin, a downstream target of BETA2/NeuroD1 in the pancreas, is involved in glucose-mediated insulin secretion. AB - BETA2 (NeuroD1) is a member of the basic helix-loop-helix transcription factor family. BETA2 plays an important role in the development of the pancreas and the nervous system. Using microarray technology, we identified neuronatin (Nnat) as differentially expressed between wild-type (WT) and knockout (KO) pancreatic RNA from embryonic day 14 (e14.5). NNAT is a member of the proteolipid family of amphipathic polypeptides and is believed to be involved in ion channel transport or channel modulation. Northern blot and in situ hybridization analysis of WT and KO samples confirmed the downregulation of Nnat in pancreas of mutant BETA2 embryos. Chromatin immunoprecipitation and gel shift assays were performed and demonstrated the presence of BETA2 on the Nnat promoter, thus confirming the direct transcriptional regulation of Nnat by BETA2. To assess NNAT potential function, we performed knockdown studies by siRNA in NIT cells and observed a reduction in the ability of the NIT cells to respond to glucose. These results suggest for the first time an important role for NNAT in insulin secretion and for proper beta-cell function. PMID- 15793248 TI - Serum- and glucocorticoid-inducible kinase 1 (SGK1) mediates glucocorticoid induced inhibition of insulin secretion. AB - Glucocorticoid excess predisposes to the development of diabetes, at least in part through impairment of insulin secretion. The underlying mechanism has remained elusive. We show here that dexamethasone upregulates transcription and expression of the serum- and glucocorticoid-inducible kinase 1 (SGK1) in insulin secreting cells, an effect reversed by mifepristone (RU486), an antagonist of the nuclear glucocorticoid receptor. When coexpressed in Xenopus oocytes, SGK1 increases the activity of voltage-gated K(+) channel K(v)1.5. In INS-1 cells, dexamethasone stimulates the transcription of K(v)1.5, increases the repolarizing outward current, reduces peak values of [Ca(2+)](i) oscillations, and decreases glucose-induced insulin release. The latter effect is reversed by K(+) channel blockers 4-aminopyridine and tetraethylammonium and by a more selective K(v)1.5 channel inhibitor MSD-D. Dexamethasone also increases expression of K(v)1.5 in mouse islets and reduces glucose-induced insulin secretion, an effect reversed by MSD-D. In islets isolated from wild-type but not SGK1 knockout mice, dexamethasone significantly blunted glucose-, forskolin-, and phorbol myristic acid-induced insulin release. In conclusion, dexamethasone stimulates the transcription of SGK1, which in turn upregulates the activity of voltage-gated K(+) channels. Increased K(+) channel activity reduces Ca(2+) entry through voltage-gated Ca(2+) channels and insulin release. PMID- 15793249 TI - The role of amylin and glucagon in the dampening of glycemic excursions in children with type 1 diabetes. AB - Postprandial hyperglycemia and preprandial hypoglycemia contribute to poor glycemic control in type 1 diabetes. We hypothesized that postprandial glycemic excursions could be normalized in type 1 diabetes by suppressing glucagon with pramlintide acetate in the immediate postprandial period and supplementing glucagon in the late postprandial period. A total of 11 control subjects were compared with 8 type 1 diabetic subjects on insulin pump therapy, using the usual insulin bolus-to-carbohydrate ratio during a standard liquid meal. Type 1 diabetic subjects were then randomized to two open-labeled studies. On one occasion, type 1 diabetic subjects received a 60% increase in the insulin bolus to-carbohydrate ratio with minidose glucagon rescue injections, and on the other occasion type 1 diabetic subjects received 30-45 microg pramlintide with their usual insulin bolus-to-carbohydrate ratio. Glucose, glucagon, amylin (pramlintide), and insulin concentrations were measured for 420 min. The plasma glucose area under the curve (AUC) for 0-420 min was lower in control versus type 1 diabetic subjects (316 +/- 5 vs. 929 +/- 18 mg x h(-1) x dl(-1), P < 0.0001). Pramlintide, but not an increase in insulin, reduced immediate postprandial hyperglycemia (AUC(0-180 min) 470 +/- 43 vs. 434 +/- 48 mg x h(-1) x dl(-1), P < 0.01). Pramlintide administration suppressed glucagon (P < 0.02), and glucagon injections prevented late hypoglycemia with increased insulin. In summary, in type 1 diabetes, glucagon modulation with pramlintide as an adjunct to insulin therapy may prove beneficial in controlling postmeal glycemic swings. PMID- 15793250 TI - Skeletal muscle lipid accumulation in type 2 diabetes may involve the liver X receptor pathway. AB - Liver X receptors (LXRs) are important regulators of cholesterol and lipid metabolism and are also involved in glucose metabolism. However, the functional role of LXRs in human skeletal muscle is at present unknown. This study demonstrates that chronic ligand activation of LXRs by a synthetic LXR agonist increases the uptake, distribution into complex cellular lipids, and oxidation of palmitate as well as the uptake and oxidation of glucose in cultured human skeletal muscle cells. Furthermore, the effect of the LXR agonist was additive to acute effects of insulin on palmitate uptake and metabolism. Consistently, activation of LXRs induced the expression of relevant genes: fatty acid translocase (CD36/FAT), glucose transporters (GLUT1 and -4), sterol regulatory element-binding protein-1c, peroxisome proliferator-activated receptor-gamma, carnitine palmitoyltransferase-1, and uncoupling protein 2 and 3. Interestingly, in response to activation of LXRs, myotubes from patients with type 2 diabetes showed an elevated uptake and incorporation of palmitate into complex lipids but an absence of palmitate oxidation to CO(2). These results provide evidence for a functional role of LXRs in both lipid and glucose metabolism and energy uncoupling in human myotubes. Furthermore, these data suggest that increased intramyocellular lipid content in type 2 diabetic patients may involve an altered response to activation of components in the LXR pathway. PMID- 15793251 TI - Loss of heparan N-sulfotransferase in diabetic liver: role of angiotensin II. AB - The basis for accelerated atherosclerosis in diabetes is unclear. Diabetes is associated with loss of heparan sulfate (HS) from the liver, which may impede lipoprotein clearance and thereby worsen atherosclerosis. To study hepatic HS loss in diabetes, we examined regulation of HS N-deacetylase/N-sulfotransferase-1 (NDST), a key enzyme in hepatic HS biosynthesis. Hepatic NDST mRNA, protein, and enzymatic activity were suppressed by >50% 2 weeks after induction of type 1 diabetes in rats. Treatment of diabetic rats with enalapril, an ACE inhibitor, had no effect on hyperglycemia or hepatic NDST mRNA levels, yet increased hepatic NDST protein and enzymatic activity. Similar results were obtained in diabetic animals treated with losartan, which blocks the type 1 receptor for angiotensin II (AngII). Consistent with these findings, diabetic livers exhibited increased ACE expression, and addition of AngII to cultured hepatoma cells reduced NDST activity and protein. We conclude that diabetes substantially suppresses hepatic NDST mRNA, protein, and enzymatic activity. AngII contributes to suppression of NDST protein and enzymatic activity, whereas mRNA suppression occurs independently. Suppression of hepatic NDST may contribute to diabetic dyslipidemia, and stimulation of NDST activity by AngII inhibitors may provide cardiovascular protection. PMID- 15793252 TI - Metformin prevents glucose-induced protein kinase C-beta2 activation in human umbilical vein endothelial cells through an antioxidant mechanism. AB - Hyperglycemia determines the vascular complications of diabetes through different mechanisms: one of these is excessive activation of the isoform beta2 of protein kinase C (PKC-beta2). Metformin, a widely used antidiabetic agent, is associated with decreased cardiovascular mortality in obese type 2 diabetic patients. Therefore, we assessed the role of metformin in glucose-induced activation of PKC beta2 and determined the mechanism of its effect in human umbilical venous endothelial cells grown to either normo- (5 mmol/l) or hyperglycemia (10 mmol/l) and moderately and acutely exposed to 25 mmol/l glucose. We studied PKC-beta2 activation by developing adenovirally expressed chimeras encoding fusion protein between green fluorescent protein (GFP) and conventional beta2 isoform (PKC-beta2 GFP). Glucose (25 mmol/l) induced the translocation of PKC-beta2-GFP from the cytosol to the membrane in cells grown to hyperglycemia but not in those grown in normal glucose medium. Metformin (20 micromol/l) prevented hyperglycemia-induced PKC-beta2-GFP translocation. We also assessed oxidative stress under the same conditions with a 4-((9-acridine-carbonyl)amino)-2,2,6,6-tetramethylpiperidin oxyl,free radical (TEMPO-9-AC) fluorescent probe. We observed significantly increased radical oxygen species production in cells grown in hyperglycemia medium, and this effect was abolished by metformin. We show that in endothelial cells, metformin inhibits hyperglycemia-induced PKC-beta2 translocation because of a direct antioxidant effect. Our data substantiate the findings of previous large intervention studies on the beneficial effect of this drug in type 2 diabetic patients. PMID- 15793253 TI - The calcium-dependent protease calpain causes endothelial dysfunction in type 2 diabetes. AB - Cardiovascular complications are the leading cause of morbidity and mortality in diabetic patients. Endothelial dysfunction with impaired endothelial nitric oxide (NO) synthase (eNOS) activity is a widely accepted cause of diabetic vasculopathy. The mechanisms of endothelial dysfunction in diabetes remain elusive, thus limiting effective therapeutic interventions. We report novel evidence demonstrating that the calcium-dependent protease calpain causes endothelial dysfunction and vascular inflammation in the microcirculation of the ZDF (Zucker diabetic fatty) rat, a genetic rat model of type 2 diabetes. We found evidence of increased calpain activity and leukocyte trafficking in the microcirculation of ZDF rats. Inhibition of calpain activity significantly attenuated leukocyte-endothelium interactions in the vasculature of ZDF rats. Expression of cell adhesion molecules in the vascular endothelium of ZDF rats was consistently increased, and it was suppressed by calpain inhibition. In vivo measurement of endothelial NO availability demonstrated a 60% decrease in NO levels in the microcirculation of diabetic rats, which was also prevented by calpain inhibition. Immunoprecipitation studies revealed calpain-dependent loss of association between eNOS and the regulatory protein heat shock protein 90. Collectively, these data provide evidence for a novel mechanism of endothelial dysfunction and vascular inflammation in diabetes. Calpains may represent a new molecular target for the prevention and treatment of diabetic vascular complications. PMID- 15793254 TI - Nonhuman primate models for diabetic ocular neovascularization using AAV2 mediated overexpression of vascular endothelial growth factor. AB - Neovascularization leads to blindness in numerous ocular diseases, including diabetic retinopathy, age-related macular degeneration, retinopathy of prematurity, and sickle cell disease. More effective and stable treatments for ocular neovascularization are needed, yet there are major limitations in the present animal models. To develop primate models of diabetic retinopathy and choroidal neovascularization, rhesus monkeys were injected subretinally or intravitreally with an adeno-associated virus (AAV)-2 vector carrying the cDNA encoding human vascular endothelial growth factor (VEGF). Overexpression of VEGF was measured by intraocular fluid sampling over time. Neovascularization was evaluated by ophthalmoscopy through angiography, optical coherence tomography, and ultimately histopathology. Overexpression of VEGF through AAV2 results in rapid development of features of diabetic retinopathy or macular edema, depending on the targeted cell type/mode of production of VEGF and diffusion of VEGF. Nonhuman primate models will be useful in testing long-term safety and efficacy of novel therapeutic agents for blinding neovascular diseases. PMID- 15793255 TI - Prevention of type 2 diabetes with troglitazone in the Diabetes Prevention Program. AB - The Diabetes Prevention Program (DPP) was a randomized clinical trial of prevention of type 2 diabetes in high-risk people. Troglitazone, an insulin sensitizing agent, was used initially but was discontinued during the trial. Troglitazone therapy was compared with other DPP interventions, considering both the short-term "in-trial" results and the longer-term results after troglitazone were discontinued. From 1996 to 1998, participants were randomly assigned to treatment with metformin (n = 587), troglitazone (n = 585), double placebo (n = 582), or intensive lifestyle intervention (ILS) (n = 589). Because of concern regarding its liver toxicity, the troglitazone arm was discontinued in June 1998, after which follow-up of all participants continued. During the mean 0.9 year (range 0.5-1.5 years) of troglitazone treatment, the diabetes incidence rate was 3.0 cases/100 person-years, compared with 12.0, 6.7, and 5.1 cases/100 person years in the placebo, metformin, and ILS participants (P < 0.001, troglitazone vs. placebo; P = 0.02, troglitazone vs. metformin; P = 0.18, troglitazone vs. ILS). This effect of troglitazone was in part due to improved insulin sensitivity with maintenance of insulin secretion. During the 3 years after troglitazone withdrawal, the diabetes incidence rate was almost identical to that of the placebo group. Troglitazone, therefore, markedly reduced the incidence of diabetes during its limited period of use, but this action did not persist. Whether other thiazolidinedione drugs used for longer periods can safely prevent diabetes remains to be determined. PMID- 15793257 TI - Inhalation of insulin (Exubera) is associated with augmented disposal of portally infused glucose in dogs. AB - The results of the present study, using the conscious beagle dog, demonstrate that inhaled insulin (INH; Exubera) provides better glycemic control during an intraportal glucose load than identical insulin levels induced by insulin (Humulin) infusion into the inferior vena cava (IVC). In the INH group (n = 13), portal glucose infusion caused arterial plasma glucose to rise transiently (152 +/- 9 mg/dl), before it returned to baseline (65 min) for the next 2 h. Net hepatic glucose uptake was minimal, whereas nonhepatic uptake rose to 12.5 +/- 0.5 mg x kg(-1) x min(-1) (65 min). In the IVC group (n = 9), arterial glucose rose rapidly (172 +/- 6 mg/dl) and transiently fell to 135 +/- 13 mg/dl (65 min) before returning to 165 +/- 15 mg/dl (125 min). Plasma glucose excursions and hepatic glucose uptake were much greater in the IVC group, whereas nonhepatic uptake was markedly less (8.6 +/- 0.9 mg x kg(-1) x min(-1); 65 min). Insulin kinetics and areas under the curve were identical in both groups. These data suggest that inhalation of Exubera results in a unique action on nonhepatic glucose clearance. PMID- 15793256 TI - Direct activation of glucose transport in primary human myotubes after activation of peroxisome proliferator-activated receptor delta. AB - Activators of peroxisome proliferator-activated receptor (PPAR)gamma have been studied intensively for their insulin-sensitizing properties and antidiabetic effects. Recently, a specific PPARdelta activator (GW501516) was reported to attenuate plasma glucose and insulin levels when administered to genetically obese ob/ob mice. This study was performed to determine whether specific activation of PPARdelta has direct effects on insulin action in skeletal muscle. Specific activation of PPARdelta using two pharmacological agonists (GW501516 and GW0742) increased glucose uptake independently of insulin in differentiated C2C12 myotubes. In cultured primary human skeletal myotubes, GW501516 increased glucose uptake independently of insulin and enhanced subsequent insulin stimulation. PPARdelta agonists increased the respective phosphorylation and expression of AMP activated protein kinase 1.9-fold (P < 0.05) and 1.8-fold (P < 0.05), of extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase (MAPK) 2.2-fold (P < 0.05) and 1.7-fold (P < 0.05), and of p38 MAPK 1.2-fold (P < 0.05) and 1.4-fold (P < 0.05). Basal and insulin-stimulated protein kinase B/Akt was unaltered in cells preexposed to PPARdelta agonists. Preincubation of myotubes with the p38 MAPK inhibitor SB203580 reduced insulin- and PPARdelta-mediated increase in glucose uptake, whereas the mitogen-activated protein kinase kinase inhibitor PD98059 was without effect. PPARdelta agonists reduced mRNA expression of PPARdelta, sterol regulatory element binding protein (SREBP)-1a, and SREBP-1c (P < 0.05). In contrast, mRNA expression of PPARgamma, PPARgamma coactivator 1, GLUT1, and GLUT4 was unaltered. Our results provide evidence to suggest that PPARdelta agonists increase glucose metabolism and promote gene regulatory responses in cultured human skeletal muscle. Moreover, we provide biological validation of PPARdelta as a potential target for antidiabetic therapy. PMID- 15793258 TI - Genetic variations in the gene encoding ELMO1 are associated with susceptibility to diabetic nephropathy. AB - To search for a gene(s) conferring susceptibility to diabetic nephropathy (DN), we genotyped over 80,000 gene-based single nucleotide polymorphisms (SNPs) in Japanese patients and identified that the engulfment and cell motility 1 gene (ELMO1) was a likely candidate for conferring susceptibility to DN, in view of the significant association of an SNP in this gene with the disease (intron 18+9170, GG vs. GA+AA, chi(2) = 19.9, P = 0.000008; odds ratio 2.67, 95% CI 1.71 4.16). In situ hybridization (ISH) using the kidney of normal and diabetic mice revealed that ELMO1 expression was weakly detectable mainly in tubular and glomerular epithelial cells in normal mouse kidney and was clearly elevated in the kidney of diabetic mice. Subsequent in vitro analysis revealed that ELMO1 expression was elevated in cells cultured under high glucose conditions (25 mmol/l) compared with cells cultured under normal glucose conditions (5.5 mmol/l). Furthermore, we identified that the expression of extracellular matrix protein genes, such as type 1 collagen and fibronectin, were increased in cells that overexpress ELMO1, whereas the expression of matrix metalloproteinases was decreased. These results indicate that ELMO1 is a novel candidate gene that both confers susceptibility to DN and plays an important role in the development and progression of this disease. PMID- 15793259 TI - The exon 1 Cys7Gly polymorphism within the betacellulin gene is associated with type 2 diabetes in African Americans. AB - In vitro and in vivo studies suggest a role for betacellulin in islet neogenesis and regeneration. Since abnormalities in beta-cell function play a role in the development of type 2 diabetes, a mutation in the betacellulin gene could potentially contribute to the development of type 2 diabetes. Using RT-PCR, we initially determined that betacellulin was expressed in 9- to 24-week-old human fetal pancreas. We then screened the betacellulin gene for mutations in subjects with type 2 diabetes and identified seven polymorphisms in segments encompassing the 5' untranslated region (G-233C, A-226G), exon 1 (TGC19GGC, Cys7Gly), exon 2 (CTC130TTC, Leu44Phe), exon 4 (TTG370ATG, Leu124Met), intron 2 (T-31C), and intron 4 (C-4T). These polymorphisms were genotyped in an expanded set of diabetic case and control subjects. Among African Americans (n = 334), the frequency of the Gly7 allele in exon 1 was 31.9% in diabetic case subjects compared with 45.1% in nondiabetic control subjects (P = 0.0004). Allele frequencies for the other polymorphisms did not differ significantly between African-American case and control subjects. Additionally, there were no significant differences in allele frequencies between case and control subjects among the Caucasian sample (n = 426) for any of the seven polymorphisms, including the Gly7 variant. Further studies will be needed to understand the different roles that betacellulin polymorphisms play in susceptibility to type 2 diabetes in Caucasians and African Americans. PMID- 15793260 TI - Genetic analysis of HNF4A polymorphisms in Caucasian-American type 2 diabetes. AB - Hepatocyte nuclear factor 4alpha (HNF4A), the gene for the maturity-onset diabetes of the young type 1 monogenic form of type 2 diabetes, is within the type 2 diabetes-linked region on chromosome 20q12-q13.1 and, consequently, is a positional candidate gene for type 2 diabetes in the general population. Previous studies have identified only a few rare coding mutations. However, recent studies suggest that single nucleotide polymorphisms (SNPs) located near the P2 (beta cell) promoter of HNF4A are associated with diabetes susceptibility. In this study, we evaluated 23 SNPs spanning 111 kb including the HNF4A gene for association with type 2 diabetes in a collection of Caucasian type 2 diabetic patients with end-stage renal disease (n = 300) and control subjects (n = 310). None of the individual SNPs were associated with type 2 diabetes in this collection of case subjects (P values ranging from 0.06 to 0.99). However, haplotype analysis identifies significant differences between haplotype frequencies in type 2 diabetic case and control subjects (P = 0.013 to P < 0.001), with two uncommon "risk" haplotypes (2.4 and 2.2% of chromosomes) and two uncommon "protective" haplotypes (7.1 and 5.0% of chromosomes) accounting for the evidence of association. Our results suggest that type 2 diabetes linked to 20q12 13 is a heterogeneous disease in which different populations may have different type 2 diabetes susceptibility loci. PMID- 15793261 TI - Ultrafine mapping of SNPs from mouse strains C57BL/6J, DBA/2J, and C57BLKS/J for loci contributing to diabetes and atherosclerosis susceptibility. AB - The inbred mouse strain C57BLKS/J (BKS) carrying a mutation of the leptin receptor lepr(-/-) (BKS-db) is a classic mouse model of type 2 diabetes. While BKS was originally presumed to be a substrain of C57BL/6J (B6), it has become apparent that its genome contains introgressed regions from a DBA/2 (DBA)-like strain and perhaps other unidentified sources. It has been hypothesized that the strikingly enhanced diabetes susceptibility of BKS-db compared with B6-db is conferred by this introgressed DNA. Using high-density single nucleotide polymorphisms, we have mapped the DBA and other contaminating DNA regions present in BKS. Thus, approximately 70% of its genome appears to derive from B6, with approximately 20% from DBA and another 9% from an unidentified donor. Comparison with 56 diverse inbred strains suggests that this donor may be a less common inbred strain or an outbred or wild strain. Using expression data from a B6 x DBA cross, we identified differentially regulated genes between these two strains. Those cis-regulated genes located on DBA-like blocks in BKS constitute primary candidates for genes contributing to diabetes susceptibility in the BKS-db strain. To further prioritize these candidates, we identified those cis-acting expression quantitative trait loci whose expression significantly correlates with diabetes-related phenotypes. PMID- 15793262 TI - An autosomal genome-wide scan for loci linked to pre-diabetic phenotypes in nondiabetic Chinese subjects from the Stanford Asia-Pacific Program of Hypertension and Insulin Resistance Family Study. AB - Type 2 diabetes is a complex disease involving both genetic and environmental components. Abnormalities in insulin secretion and insulin action usually precede the development of type 2 diabetes and can serve as good quantitative measures for genetic mapping. We therefore undertook an autosomal genomic search to locate the quantitative trait locus (QTL) linked to these traits in 1,365 nondiabetic Chinese subjects from 411 nuclear families. Residuals of these log-transformed quantitative traits were analyzed in multipoint linkage analysis using a variance components approach. The most significant QTL for fasting insulin, which coincides with the QTL for homeostasis model assessment of insulin resistance, was located at 37 cM on chromosome 20, with a maximum empirical logarithm of odds (LOD) score of 3.01 (empirical P = 0.00006) when adjusted for age, sex, BMI, antihypertensive medications, recruitment centers, and environmental factors. In the same region, a QTL for fasting glucose was identified at 51 cM, with an empirical LOD score of 2.03 (empirical P = 0.0012). There were other loci with maximum empirical LOD scores >or=1.29 located on chromosomes 1q, 2p, 5q, 7p, 9q, 10p, 14q, 18q, and 19q for different diabetes-related traits. These loci may harbor genes that regulate glucose homeostasis either independently or via interactions of the genes within these regions. PMID- 15793263 TI - ENPP1/PC-1 K121Q polymorphism and genetic susceptibility to type 2 diabetes. AB - Genetic susceptibility modulates the impact of obesity on risk for type 2 diabetes. The present study evaluates the role of ENPP1 K121Q polymorphism in prediction of type 2 diabetes in three populations that differ in susceptibility to diabetes and environmental exposure. The three cohorts included 679 nonmigrant South Asians living in Chennai, India (223 with type 2 diabetes); 1,083 migrant South Asians living in Dallas, Texas (121 with type 2 diabetes); and 858 nonmigrant Caucasians living in Dallas, Texas (141 with type 2 diabetes). Patients with type 2 diabetes were included in these cohorts if they had diabetes onset before the age of 60 years. The prevalence of subjects carrying the polymorphic ENPP1 121Q allele was 25% in the nondiabetic group and 34% in the diabetic group of South Asians living in Chennai (P = 0.01). The prevalence in the nondiabetic and diabetic groups were 33 and 45% (P = 0.01) for the South Asians living in Dallas and 26 and 39% (P = 0.003) for the Caucasians. Although further replication studies are necessary to test the validity of the described genotype-phenotype relationship, our study supports the hypothesis that ENPP1 121Q predicts genetic susceptibility to type 2 diabetes in both South Asians and Caucasians. PMID- 15793264 TI - A single nucleotide polymorphism in MGEA5 encoding O-GlcNAc-selective N-acetyl beta-D glucosaminidase is associated with type 2 diabetes in Mexican Americans. AB - Excess O-glycosylation of proteins by O-linked beta-N-acetylglucosamine (O GlcNAc) may be involved in the pathogenesis of type 2 diabetes. The enzyme O GlcNAc-selective N-acetyl-beta-d glucosaminidase (O-GlcNAcase) encoded by MGEA5 on 10q24.1-q24.3 reverses this modification by catalyzing the removal of O GlcNAc. We have previously reported the linkage of type 2 diabetes and age at diabetes onset to an overlapping region on chromosome 10q in the San Antonio Family Diabetes Study (SAFADS). In this study, we investigated menangioma expressed antigen-5 (MGEA5) as a positional candidate gene. Twenty-four single nucleotide polymorphisms (SNPs), identified by sequencing 44 SAFADS subjects, were genotyped in 436 individuals from 27 families whose data were used in the original linkage report. Association tests indicated significant association of a novel SNP with the traits diabetes (P = 0.0128, relative risk = 2.77) and age at diabetes onset (P = 0.0017). The associated SNP is located in intron 10, which contains an alternate stop codon and may lead to decreased expression of the 130 kDa isoform, the isoform predicted to contain the O-GlcNAcase activity. We investigated whether this variant was responsible for the original linkage signal. The variance attributed to this SNP accounted for approximately 25% of the logarithm of odds. These results suggest that this variant within the MGEA5 gene may increase diabetes risk in Mexican Americans. PMID- 15793265 TI - Variation in the gene for muscle-specific AMP deaminase is associated with insulin clearance, a highly heritable trait. AB - The rising prevalence of the insulin resistance syndrome in our society necessitates a better understanding of the genetic determinants of all aspects of insulin action and metabolism. We evaluated the heritability of insulin sensitivity and the metabolic clearance rate of insulin (MCRI) as quantified by the euglycemic-hyperinsulinemic clamp in 403 Mexican Americans. We tested the candidate gene AMP deaminase 1 (AMPD1) for association with insulin-related traits because it codes for an enzyme that has the potential to influence multiple aspects of insulin pharmacodynamics. By converting AMP to inosine monophosphate, AMPD1 plays a major role in regulating cellular AMP levels; AMP activates AMP kinase, an enzyme that modulates cellular energy and insulin action. We determined that nine AMPD1 single nucleotide polymorphisms (SNPs) defined two haplotype blocks. Insulin clearance was found to have a higher heritability (h(2) = 0.58) than fasting insulin (h(2) = 0.38) or insulin sensitivity (h(2) = 0.44). The MCRI was associated with AMPD1 SNPs and haplotypes. Insulin clearance is a highly heritable trait, and specific haplotypes within the AMPD1 gene, which encodes a skeletal muscle-specific protein, are associated with variation in insulin clearance. We postulated that the processes of insulin action and insulin clearance in skeletal muscle are highly regulated and that AMPD1 function may play an important role in these phenomena. PMID- 15793266 TI - Association of the diabetes gene calpain-10 with subclinical atherosclerosis: the Mexican-American Coronary Artery Disease Study. AB - The powerful relation between atherosclerosis and diabetes may have a common genetic basis. However, few genes predisposing to both have been identified. Calpain-10 (CAPN10) was the first gene for type 2 diabetes identified by positional cloning, wherein a combination of haplotypes conferred increased risk of diabetes. We sought to determine whether CAPN10 influences subclinical atherosclerosis. Among nondiabetic subjects from 85 Mexican-American families with a history of coronary artery disease, subclinical atherosclerosis was assessed by common carotid artery intima-media thickness (IMT), insulin sensitivity was assessed by hyperinsulinemic-euglycemic clamp, and insulin secretion was estimated by the oral glucose tolerance test. These phenotypes were tested for association with CAPN10 haplotypes. Haplotype 1112 (of single nucleotide polymorphisms [SNPs] 44, 43, 56, and 63) was associated with increased IMT, while haplotype 1221 was associated with decreased IMT. The 112/121 haplotype combination (of SNPs 43, 56, and 63), originally found to confer increased risk for diabetes, was associated with the largest IMT in our study population. CAPN10 was also associated with both insulin sensitivity and insulin secretion. Covariate analysis suggested that CAPN10 affects IMT independently of these diabetes-related phenotypes. The fact that the diabetes gene CAPN10 also influences the risk for atherosclerosis shows that inherited factors may underlie the frequent co-occurrence of these two conditions. PMID- 15793267 TI - The rat diabetes susceptibility locus Iddm4 and at least one additional gene are required for autoimmune diabetes induced by viral infection. AB - BBDR rats develop autoimmune diabetes only after challenge with environmental perturbants. These perturbants include polyinosinic:polycytidylic acid (poly I:C, a ligand of toll-like receptor 3), agents that deplete regulatory T-cell (Treg) populations, and a non-beta-cell cytopathic parvovirus (Kilham rat virus [KRV]). The dominant diabetes susceptibility locus Iddm4 is required for diabetes induced by treatment with poly I:C plus Treg depletion. Iddm4 is penetrant in congenic heterozygous rats on the resistant WF background and is 79% sensitive and 80% specific as a predictor of induced diabetes. Surprisingly, an analysis of 190 (BBDR x WF)F2 rats treated with KRV after brief exposure to poly I:C revealed that the BBDR-origin allele of Iddm4 is necessary but not entirely sufficient for diabetes expression. A genome scan identified a locus on chromosome 17, designated Iddm20, that is also required for susceptibility to diabetes after exposure to KRV and poly I:C (logarithm of odds score 3.7). These data suggest that the expression of autoimmune diabetes is a complex process that requires both major histocompatibility complex genes that confer susceptibility and additional genes such as Iddm4 and Iddm20 that operate only in the context of specific environmental perturbants, amplifying the immune response and the rate of disease progression. PMID- 15793269 TI - Is puberty an accelerator of type 1 diabetes in IL6-174CC females? AB - The pubertal peak in onset of type 1 diabetes occurs earlier in girls than boys. We postulated that this sex difference might be mediated in part by estrogen or by genes regulated by estrogen, such as the interleukin-6 (IL6) gene. Previous studies concerning the role of an estrogen-sensitive single nucleotide polymorphism (SNP) in the IL6 promoter in type 1 diabetes have proved contradictory. We therefore selected a large, genetically homogenous population based cohort, analyzed by age at onset and sex, to test the hypothesis that the IL6-174G>C SNP affects age at onset of type 1 diabetes in females but not in males. We found that the IL6-174CC genotype was significantly less frequent in females diagnosed after than in those diagnosed before the age of 10 years (19 vs. 13%, P = 0.016). No genotype difference was observed in males stratified for age at onset. Among children diagnosed after age 10, the median age of onset was 11.9 years (intraquartile range 10.7-14.6) in 34 girls homozygous for IL6-174C compared with 13.2 years (11.6-15.4) in 229 girls with other genotypes and 13.5 years (12.0-15.6) in 339 males with any IL6-174 genotype (P = 0.012). These data support the hypothesis that pubertal changes may contribute to accelerated onset of type 1 diabetes in genetically susceptible females. This phenomenon may be orchestrated by the action of estrogen on the IL6 promoter. PMID- 15793268 TI - Genetic variation at the ACE gene is associated with persistent microalbuminuria and severe nephropathy in type 1 diabetes: the DCCT/EDIC Genetics Study. AB - The development and progression of microvascular complications have been extensively documented in a cohort of type 1 diabetic subjects enrolled in the Diabetes Control and Complications Trial (DCCT) and followed in the Epidemiology of Diabetes Interventions and Complications (EDIC) study. We describe the association of genetic variation in the ACE gene in 1,365 DCCT/EDIC subjects with incident persistent microalbuminuria (n = 312) and severe nephropathy (n = 115). We studied three markers (rs1800764, insertion/deletion, and rs9896208) in the ACE gene that allowed us to capture genetic variation in the common haplotypes occurring at frequencies of >5% in Caucasians. Compared with the more frequent genotype (D/I) for the insertion/deletion polymorphism, in multivariate models, the I/I genotype conferred a lower risk for persistent microalbuminuria (hazard ratio [HR] 0.62 [95% CI 0.43-0.89], P = 0.009) and severe nephropathy (0.56 [0.32 0.96], P = 0.033). Variation at the two other markers, rs1800764 and rs9896208, were also associated with these renal outcomes. In addition, homozygosity for the common haplotype TIC (which corresponded to the T, insertion, and C alleles at the three markers, rs1800764, insertion/deletion, and rs9896208, respectively) versus the CDT/TIC haplotype pair was associated with lower risk for development of persistent microalbuminuria (HR 0.49 [0.32-0.75], P = 0.0009) and severe nephropathy (0.41 [0.22-0.78], P = 0.006). Our findings in the DCCT/EDIC cohort provide strong evidence that genetic variation at the ACE gene is associated with the development of nephropathy in patients with type 1 diabetes. PMID- 15793270 TI - Theorizing about systems: an ecological task for patient safety research. PMID- 15793271 TI - Assisting older adults to communicate their postoperative pain. AB - An intervention assisting older adults to communicate their pain was tested in a posttest-only experiment. Thirty-eight preoperative older adults were randomly assigned to a communication group watching a videotape about communicating and managing postoperative pain or a comparison group watching a videotape about managing postoperative pain only. Pain was measured on Postoperative Days 1 and 2, and 1 and 7 days after hospital discharge by a data collector blind to the condition. The communication group reported greater pain relief and less pain interference on Postoperative Day 1. The comparison group reported greater pain relief on Postoperative Day 2 after attaining a pain interference level similar to the pain communication group. The pain communication intervention had modest effects for reducing pain interference with activities on Postoperative Day 1. Greater pain relief might be achieved when older adults and their health care providers are more knowledgeable about both pain communication and pain management. PMID- 15793272 TI - Cognitive-behavioral treatment combined with asthma education for adults with asthma and coexisting panic disorder. AB - This study tested the efficacy of a nurse-administered 8-week group treatment program for adults with asthma suffering from coexisting panic disorder. The program consisted of cognitive behavioral treatment (CBT) for panic disorder combined with asthma education (AE). Forty-eight women with a confirmed diagnosis of asthma and panic disorder were randomly allocated to a treatment condition (n=25) and a wait-list control condition (n=23). Twenty-five participants--15 in the treatment group and 10 in the wait-list control group--completed treatment. Repeated measures ANOVA procedures were used to compare the groups on panic and asthma outcomes at posttreatment and 6-month follow-up. The results demonstrate that the CBT-AE program is capable of producing substantial and durable antipanic and antianxiety treatment effects and led to substantial but nonsustained improvement in morning peak-flow expiratory rate and asthma-related quality of life. Implications of these findings for this clinical population are addressed. PMID- 15793273 TI - The role of spirituality in holistic weight management. AB - This descriptive, feasibility study was designed to determine how weight management patients defined spirituality and its connection with weight management. Relationships among spirituality assessment, spiritual well-being, self-esteem, and quality of life were explored. This study arose from clinical observations of possible relationships among patients' weight management failures, negative beliefs about self, and spiritual distress. Participants were 34 of 104 adult potential participants from a holistic weight management clinical practice. Survey data were analyzed using qualitative content analysis and quantitative linear regression analyses. Participants readily defined spirituality; significant linear relationships were found: Total spiritual well being explained approximately 47% of the variance for self-esteem and existential spiritual well-being accounted for approximately 68% of the variance for self esteem. Similarly, existential spiritual well-being explained approximately 35% of the variance of quality of life. For this convenience sample, spiritual well being was significantly related to self-esteem and quality of life. PMID- 15793274 TI - Happiness as related to gender and health in early adolescents. AB - The purposes of this study were to examine gender differences in happiness; to examine the relationship between happiness and several health variables, such as perceived health status, clinical health, and wellness, in early adolescents; and to examine further these relationships for boys and girls separately. The final sample consisted of 151 early adolescent boys and girls who were in the seventh and eighth grades of an urban middle school. Students responded to instruments measuring happiness, perceived health status, clinical health, and wellness. Results indicated there were no gender differences in happiness between boys and girls. Statistically significant positive correlations were found between happiness and the health-related variables for the entire sample, with different patterns of relationships when boys and girls were analyzed separately. The application of findings to nursing is discussed. PMID- 15793275 TI - Testing videotape education for heart failure. AB - This pilot study tested a videotape intervention designed to improve patient self management of heart failure (HF). Content of the video series (produced professionally under a federal grant) is based on national, scientifically validated guidelines for HF home management. Outcomes tested were HF knowledge, symptom reporting, and functional status. Participants were 10 newly diagnosed HF patients (mean age 67). After viewing the tapes, data indicated participants had a clinically relevant improvement in HF knowledge, and improved or maintained HF health status. None were rehospitalized during the 60-day follow-up period. One patient contacted his/her physician to report weight gain, as prompted by the videotapes. The cost data indicated that patients paid $177 out of pocket monthly for medications and all were low income. These results indicate the need for further testing of the videotape as a potentially cost-effective method of teaching about HF self-management and daily home self-monitoring. PMID- 15793276 TI - Tyrosine phosphorylation and endothelial cell barrier regulation. PMID- 15793277 TI - Mechanogenomic control of DNA exposure and sequestration. PMID- 15793278 TI - Transgenic sickle mice are markedly sensitive to renal ischemia-reperfusion injury. AB - Ischemic injury is invoked as a mechanism contributing to end-organ damage and other complications of sickle cell disease (SCD). However, the intrinsic sensitivity of tissues in SCD to ischemic insults has never been addressed. We examined the effect of renal ischemia in a transgenic mouse expressing human sickle hemoglobin. Twenty-four hours after bilateral, total renal artery occlusion for 15 minutes, transgenic sickle mice exhibited worse renal function and more marked histological injury. With bilateral renal ischemia of greater duration (22.5 minutes), and after 6 hours, transgenic sickle mice exhibited massive vascular congestion, sickling of red blood cells, more marked histological injury in the kidney, and more prominent congestion in the capillary beds in the lungs and heart. Additionally, serum amyloid P-component, the murine homologue of C-reactive protein, was markedly increased in transgenic sickle mice as compared to wild-type mice. Twenty-four hours after bilateral renal ischemia for 22.5 minutes, transgenic sickle mice exhibited 28% mortality, with no mortality observed in any other group. With bilateral renal ischemia of short or long duration, renal expression of caspase-3 was most prominent in transgenic sickle mice subjected to ischemia. Thus, renal ischemia in this murine model induces more severe renal injury and extrarenal complications. We conclude that tissues in SCD exhibit heightened vascular congestion and sensitivity to ischemia and that clinically apparent or silent episodes of ischemia may contribute to the complications of SCD. PMID- 15793279 TI - Plasma protein haptoglobin modulates renal iron loading. AB - Haptoglobin is the plasma protein with the highest binding affinity for hemoglobin. The strength of hemoglobin binding and the existence of a specific receptor for the haptoglobin-hemoglobin complex in the monocyte/macrophage system clearly suggest that haptoglobin may have a crucial role in heme-iron recovery. We used haptoglobin-null mice to evaluate the impact of haptoglobin gene inactivation on iron metabolism. Haptoglobin deficiency led to increased deposition of hemoglobin in proximal tubules of the kidney instead of the liver and the spleen as occurred in wild-type mice. This difference in organ distribution of hemoglobin in haptoglobin-deficient mice resulted in abnormal iron deposits in proximal tubules during aging. Moreover, iron also accumulated in proximal tubules after renal ischemia-reperfusion injury or after an acute plasma heme-protein overload caused by muscle injury, without affecting morphological and functional parameters of renal damage. These data demonstrate that haptoglobin crucially prevents glomerular filtration of hemoglobin and, consequently, renal iron loading during aging and following acute plasma heme protein overload. PMID- 15793280 TI - A hot new twist to hair biology: involvement of vanilloid receptor-1 (VR1/TRPV1) signaling in human hair growth control. AB - The vanilloid receptor-1 (VR1, or transient receptor potential vanilloid-1 receptor, TRPV1) is activated by capsaicin, the key ingredient of hot peppers. TRPV1 was originally described on sensory neurons as a central integrator of various nociceptive stimuli. However, several human skin cell populations are also now recognized to express TRPV1, but with unknown function. Exploiting the human hair follicle (HF) as a prototypic epithelial-mesenchymal interaction system, we have characterized the HF expression of TRPV1 in situ and have examined TRPV1 signaling in organ-cultured human scalp HF and outer root sheath (ORS) keratinocytes in vitro. TRPV1 immunoreactivity was confined to distinct epithelial compartments of the human HF, mainly to the ORS and hair matrix. In organ culture, TRPV1 activation by capsaicin resulted in a dose-dependent and TRPV1-specific inhibition of hair shaft elongation, suppression of proliferation, induction of apoptosis, premature HF regression (catagen), and up-regulation of intrafollicular transforming growth factor-beta(2). Cultured human ORS keratinocytes also expressed functional TRPV1, whose stimulation inhibited proliferation, induced apoptosis, elevated intracellular calcium concentration, up-regulated known endogenous hair growth inhibitors (interleukin-1beta, transforming growth factor-beta(2)), and down-regulated known hair growth promoters (hepatocyte growth factor, insulin-like growth factor-I, stem cell factor). These findings strongly support TRPV1 as a significant novel player in human hair growth control, underscore the physiological importance of TRPV1 in human skin beyond nociception, and identify TRPV1 as a promising, novel target for pharmacological manipulations of epithelial growth disorders. PMID- 15793281 TI - Heparanase regulates murine hair growth. AB - Heparanase is an endoglycosidase that cleaves heparan sulfate, the main polysaccharide component of the extracellular matrix. Heparan sulfate moieties are responsible for the extracellular matrix barrier function, as well as for sequestration of heparin-binding growth factors in the extracellular matrix. Degradation of heparan sulfate by heparanase enables cell movement through extracellular barriers and releases growth factors from extracellular matrix depots, making them bioavailable. Here, we demonstrate a highly coordinated temporospatial pattern of heparanase expression and enzymatic activity during hair follicle cycling. This pattern paralleled the route and timing of follicular stem cell progeny migration and reconstitution of the lower part of the follicle, which is a prerequisite for hair shaft formation. By monitoring in vivo activation of luciferase reporter gene driven by heparanase promoter, we observed activation of heparanase gene transcription at a specific stage of the hair cycle. Heparanase was produced by rat vibrissa bulge keratinocytes, closely related to a follicular stem cell population. Heparanase contributed to the ability of the bulge-derived keratinocytes to migrate through the extracellular matrix barrier in vitro. In heparanase-overexpressing transgenic mice, increased levels of heparanase enhanced active hair growth and enabled faster hair recovery after chemotherapy-induced alopecia. Collectively, our results identify heparanase as an important regulator of hair growth and suggest that cellular mechanisms of its action involve facilitation of follicular stem cell progeny migration and release of extracellular matrix-resident, heparin-bound growth factors, thus regulating hair cycle. PMID- 15793283 TI - Growth inhibition and apoptosis in liver myofibroblasts promoted by hepatocyte growth factor leads to resolution from liver cirrhosis. AB - Liver cirrhosis is characterized by hepatic dysfunction with extensive accumulation of fibrous tissue in the liver. In response to chronic hepatic injury, hepatic portal myofibroblasts and activated hepatic stellate cells (HSCs) play a role in liver fibrosis. Although administration or gene expression of hepatocyte growth factor (HGF) leads to improvement in hepatic fibrosis/cirrhosis, the related mechanisms are not fully understood. We investigated mechanisms involved in resolution from liver cirrhosis by HGF, focusing on growth regulation and apoptosis in portal myofibroblasts. Cultured rat HSCs could not proliferate, were withdrawn after passage, and were replaced by proliferating portal myofibroblasts during the passages. In quiescent HSCs, c Met receptor expression was undetected whereas c-Met receptor expression was detected in activated HSCs and liver myofibroblasts expressing alpha-smooth muscle actin (alpha-SMA), suggesting that activated HSCs and portal myofibroblasts are targets of HGF. For cultured rat portal myofibroblasts, HGF counteracted phosphorylation of extracellular signal-regulated kinase (Erk) 1/2 and mitogenic stimulus induced by platelet-derived growth factor, induced c-jun N terminal kinase (JNK) 1 phosphorylation, and promoted apoptotic cell death. In the dimethylnitrosamine rat model of liver cirrhosis, administration of HGF suppressed proliferation while promoting apoptosis of alpha-SMA-positive cells in the liver, events that were associated with reduced hepatic expressions of alpha SMA and histological resolution from liver cirrhosis. Growth inhibition and enhanced apoptosis in portal myofibroblasts by HGF are newly identified mechanisms aiding resolution from liver fibrosis/cirrhosis by HGF. PMID- 15793282 TI - Amphiregulin and epidermal hyperplasia: amphiregulin is required to maintain the psoriatic phenotype of human skin grafts on severe combined immunodeficient mice. AB - Overexpression of amphiregulin has been shown to induce psoriasiform changes in the skin of transgenic mice shortly after birth. Therefore, amphiregulin has been suggested as a target for anti-psoriatic therapy. To test this theory, a humanized monoclonal antibody capable of neutralizing human amphiregulin was examined for anti-proliferative effects in the human skin-severe combined immunodeficient (SCID) mouse transplant model. The anti-amphiregulin antibody reduced epidermal thickness of transplanted psoriatic skin and also inhibited the hyperplastic response that developed in nonpsoriatic skin after transplantation. The same antibody also suppressed keratinocyte proliferation in monolayer culture in a dose-dependent manner. Under the same conditions in which keratinocyte proliferation was inhibited, the antibody had little effect on proliferation of human dermal fibroblasts and no effect on type I procollagen production by these cells. Taken together, these data indicate an important role for amphiregulin in psoriatic hyperplasia and suggest that inhibition of amphiregulin activity could be an efficacious therapeutic strategy for psoriasis. These data also suggest that the hyperplastic response occurring in nonpsoriatic human skin on transplantation to the SCID mouse is mediated, in large part, by amphiregulin. PMID- 15793285 TI - Hepatic gene expression in protoporphyic Fech mice is associated with cholestatic injury but not a marked depletion of the heme regulatory pool. AB - BALB/c Fech(m1Pas) mice have a mutated ferrochelatase gene resulting in protoporphyria that models the hepatic injury occurring sporadically in human erythropoietic protoporphyria. We used this mouse model to study the development of the injury and to compare the dysfunction of heme synthesis with hepatic gene expression of liver metabolism, oxidative stress, and cellular injury/inflammation. From an early age expression of total cytochrome P450 and many of its isoforms was significantly lower than in wild-type mice. However, despite massive accumulation of protoporphyrin in the liver, expression of the main genes controlling heme synthesis and catabolism (Alas1 and Hmox1, respectively) were only modestly affected even in the presence of the cytochrome P450-inducing CAR agonist 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene. In contrast, in BALB/c mice exhibiting griseofulvin-induced hepatic protoporphyria with induction and destruction of cytochrome P450, both Alas1 and Hmox1 genes were markedly up-regulated. Other expression profiles in BALB/c Fech(m1Pas) mice identified roles for oxidative mechanisms in liver injury while modulated gene expression of hepatocyte transport proteins and cholesterol and bile acid synthesis illustrated the development of cholestasis. Subsequent inflammation and cirrhosis were also shown by the up-regulation of cytokine, cell cycling, and procollagen genes. Thus, gene expression profiles studied in Fech(m1Pas) mice may provide candidates for human polymorphisms that explain the sporadic hepatic consequences of erythropoietic protoporphyria. PMID- 15793284 TI - Transforming growth factor-beta and platelet-derived growth factor signal via c Jun N-terminal kinase-dependent Smad2/3 phosphorylation in rat hepatic stellate cells after acute liver injury. AB - After liver injury, transforming growth factor-beta (TGF-beta) and platelet derived growth factor (PDGF) regulate the activation of hepatic stellate cells (HSCs) and tissue remodeling. Mechanisms of PDGF signaling in the TGF-beta triggered cascade are not completely understood. TGF-beta signaling involves phosphorylation of Smad2 and Smad3 at linker and C-terminal regions. Using antibodies to distinguish Smad2/3 phosphorylated at linker regions from those phosphorylated at C-terminal regions, we investigated Smad2/3-mediated signaling in rat liver injured by CCl(4) administration and in cultured HSCs. In acute liver injury, Smad2/3 were transiently phosphorylated at both regions. Although linker-phosphorylated Smad2 remained in the cytoplasm of alpha-smooth muscle actin-immunoreactive mesenchymal cells adjacent to necrotic hepatocytes in centrilobular areas, linker-phosphorylated Smad3 accumulated in the nuclei. c-Jun N-terminal kinase (JNK) in the activated HSCs directly phosphorylated Smad2/3 at linker regions. Co-treatment of primary cultured HSCs with TGF-beta and PDGF activated the JNK pathway, subsequently inducing endogenous linker phosphorylation of Smad2/3. The JNK pathway may be involved in migration of resident HSCs within the space of Disse to the sites of tissue damage because the JNK inhibitor SP600125 inhibited HSC migration induced by TGF-beta and PDGF signals. Moreover, treatment of HSCs with both TGF-beta and PDGF increased transcriptional activity of plasminogen activator inhibitor-1 through linker phosphorylation of Smad3. In conclusion, TGF-beta and PDGF activate HSCs by transmitting their signals through JNK-mediated Smad2/3 phosphorylation at linker regions, both in vivo and in vitro. PMID- 15793287 TI - BRAF mutations in aberrant crypt foci and hyperplastic polyposis. AB - Patients with hyperplastic polyposis have multiple hyperplastic polyps (HPs) and increased risk of colorectal carcinomas. Aberrant crypt foci (ACF) are postulated to be the earliest precursor lesions in colorectal carcinogenesis. We evaluated BRAF mutations by DNA sequencing in 53 ACF from patients with sporadic colorectal carcinomas and familial adenomatous polyposis, in 18 sporadic HPs from patients with resected colorectal cancer, and in 70 HPs, 4 serrated adenomas, 3 admixed hyperplastic-adenomatous polyps, 10 tubular adenomas, and 6 carcinomas from 17 patients with multiple/large HPs and/or hyperplastic polyposis. BRAF mutation status was compared with clinicopathological features and other genetic alterations by marginal logistic regression. BRAF mutation was present in only 2% of ACF and 6% of sporadic HPs. In contrast, BRAF mutation was present in 43% of HPs (P = 0.01 versus sporadic HPs), 75% of serrated adenomas, 33% of admixed hyperplastic-adenomatous polyps, 30% of tubular adenomas, and 33% of carcinomas from patients with multiple/large HPs and/or hyperplastic polyposis. BRAF mutation status in patients with multiple/large HPs and/or hyperplastic polyposis correlated with HPs from the same patient (odds ratio, 5.8; P = 0.0002) but associated with younger age (odds ratio, 0.83; P = 0.006 compared to older age), with a large HP (odds ratio, 22.5; P = 0.01 compared with patients with multiple HPs), with location of HPs in the right colon (odds ratio, 3.0; P = 0.03), and with methylation of the p16 gene and the MINT31 locus [odds ratio, 12.2 (P = 0.0001) and 4.4 (P = 0.02), respectively]. Our study shows that BRAF mutation status is heterogeneous among patients with multiple/large HPs and/or hyperplastic polyposis, suggesting differences in pathogenesis of HPs that indicate subsets within this phenotype. PMID- 15793286 TI - Altered epithelial cell lineage allocation and global expansion of the crypt epithelial stem cell population are associated with ileitis in SAMP1/YitFc mice. AB - Crohn's disease is characterized by cycles of mucosal injury and ulceration followed by epithelial regeneration and restoration of normal epithelial function. In this study, we examined whether ileitis in SAMP1/YitFc mice, a recombinant-inbred line that spontaneously develops ileitis resembling human Crohn's disease, was associated with alterations in normal patterns of epithelial differentiation or changes in epithelial regeneration after experimental injury. Increased numbers of Paneth, goblet, and intermediate cells were present focally in the ileum of SAMP1/YitFc mice by 4 weeks of age, before any histological evidence of acute or chronic inflammation. This increase in secretory cells became more pronounced at sites of ileitis with increasing age and inflammation. Additionally, there was mispositioning of Paneth and intermediate cells along the crypt-to-villus unit. A concomitant reduction in the number of absorptive enterocytes was observed. In contrast to the ileal-specific changes in lineage allocation, crypt stem cell numbers began to increase in both the ileum and proximal jejunum at the onset of inflammation in SAMP1/YitFc mice. These data suggest that the alterations in epithelial cell differentiation and increases in the size of the crypt stem cell population observed in SAMP1/YitFc mice are regulated by distinct mechanisms. We speculate that these epithelial alterations may play a role in the pathogenesis of ileitis in this murine model of Crohn's disease. PMID- 15793288 TI - Transdifferentiation of mature rat hepatocytes into bile duct-like cells in vitro. AB - We investigated the mechanism of phenotypic plasticity of hepatocytes in a three dimensional organoid culture system, in which hepatocytic spheroids were embedded within a collagen gel matrix. Hepatocytes expressed several bile duct markers including cytokeratin (CK) 19 soon after culture and underwent branching morphogenesis within the matrix in the presence of insulin and epidermal growth factor. Cultured hepatocytes did not express Delta-like, a specific marker for oval cells and hepatoblasts. Furthermore, hepatocytes isolated from c-kit mutant rats (Ws/Ws), which are defective in proliferation of oval cells, showed essentially the same phenotypic changes as those isolated from control rats. The bile duct-like differentiation of hepatocytes was associated with increased expression of Jagged1, Jagged2, Notch1, and several Notch target genes. CK19 expression and branching morphogenesis were inhibited by dexamethasone, a mitogen activated protein kinase kinase 1 (MEK1) inhibitor (PD98059), and a phosphatidyl inositol 3-kinase inhibitor (LY294002). After being cultured for more than 3 weeks within the gels, hepatocytes transformed into ductular structures surrounded by basement membranes. Our results suggest that hepatocytes might have the potential to transdifferentiate into bile duct-like cells without acquiring a stem-like phenotype and that this is mediated through specific protein tyrosine phosphorylation pathways. PMID- 15793290 TI - The contribution of the Fas/FasL apoptotic pathway in ulcer formation during Leishmania major-induced cutaneous Leishmaniasis. AB - Cutaneous leishmaniasis (CL), caused by the intracellular protozoan Leishmania major, is characterized by lesion formation and ulceration at the site of infection. The mechanism of ulcer formation during CL is not fully understood. The expression of Fas and FasL and the levels of apoptosis in skin biopsies and in restimulated blood mononuclear cells from patients with 1 to 7 months of L. major-induced CL were analyzed using immunohistochemistry and fluorescence activated cell sorting analysis. The levels of soluble Fas and FasL were also analyzed by enzyme-linked immunosorbent assay. A substantial number of apoptotic keratinocytes were observed mainly in the superficial epidermis of morphologically active and healing CL skin samples. Fas expression was increased on epidermis in active CL, whereas Fas expression was similar in healing and healthy epidermis. FasL-expressing macrophages and T cells were found in subepidermal infiltrate, mainly in active disease. When CL peripheral blood mononuclear cells were restimulated with L. major, Fas was up-regulated on effector T cells, and high levels of sFasL were secreted. Supernatants from restimulated cultures induced apoptosis in human keratinocytes (HaCaT), possibly through Fas/FasL interactions. Our results indicate that FasL-expressing effector T cells and macrophages may act to induce apoptosis and ulcer formation in Fas expressing keratinocytes during L. major infection. PMID- 15793289 TI - Role of lipopolysaccharide and cecal ligation and puncture on blood coagulation and inflammation in sensitive and resistant mice models. AB - The hemostatic system is severely disturbed during endotoxemia, leading to a hypercoagulable state. However, it remains uncertain to what extent hypercoagulability is the critical factor in determining the clinical course rather than just the consequence of a severe systemic inflammatory response. To answer this question, we evaluated the evolution of hemostatic and inflammatory markers, as well as histological features, in mice sensitive and resistant to two models of endotoxemia: lipopolysaccharide-injection and cecal ligation puncture. Genetic (knockout mice) and pharmacological (PJ34) blockade of the nuclear enzyme PARP-1 was used to achieve resistance to the endotoxemia. In both models, endotoxemia resulted in antithrombin deficiency, decreased platelets, and fibrin deposition in organs, which were similar in all groups of mice. By contrast, proinflammatory mediators, inflammatory cell infiltration (especially that mediated by mononuclear cells), and organ degeneration were more intense in sensitive animals. Further studies supported a negative role for the triggering of the coagulation cascade in the mortality associated with the endotoxic shock. Hirudin had a minor effect on cell infiltration and organ damage, despite causing a potent inhibition of fibrin deposition. On the other hand, a sublethal dose of lipopolysaccharide yielded significant fibrin deposition but weak activation of the inflammatory response. Our results suggest that activation of coagulation by endotoxemia is severe and independent of the inflammatory response. However, such activation may act with fibrin deposition to have a minor influence on survival in sepsis. PMID- 15793291 TI - Contribution of transmembrane tumor necrosis factor to host defense against Mycobacterium bovis bacillus Calmette-guerin and Mycobacterium tuberculosis infections. AB - To study the specific role of transmembrane tumor necrosis factor (TmTNF) in host defense mechanisms against bacillus Calmette-Guerin (BCG) and Mycobacterium tuberculosis infections, we compared the immune responses of TNF/lymphotoxin (LT) alpha(-/-) mice expressing a noncleavable transgenic TmTNF (TmTNF tg) to those of TNF/LT-alpha(-/-) and wild-type mice. Susceptibility of TNF/LT-alpha(-/-) mice to BCG infection was associated with impaired induction of systemic RANTES but not of monocyte chemoattractant protein 1 (MCP-1), the development of excessive local and systemic Th1-type immune responses, and a substantially reduced inducible nitric oxide synthase (iNOS) activity. Resistance of TmTNF tg mice to BCG infection was associated with efficient activation of iNOS in granulomas and with the regulated release of local and systemic chemokines and Th1-type cytokines. However, M. tuberculosis infection of TmTNF tg mice resulted in longer survival and enhanced resistance compared to TNF/LT-alpha(-/-) mice but higher sensitivity than wild-type mice. TmTNF tg mice exhibited reduced pulmonary iNOS expression and showed an exacerbated cellular infiltration in the lungs despite a modest bacillary content. Our data thus indicate a role for TmTNF in host defense against mycobacteria by contributing to induction and regulation of Th1-type cytokine and chemokine expression leading to development of bactericidal granulomas expressing iNOS, which critically determines susceptibility versus resistance of the host to mycobacterial infections. PMID- 15793292 TI - Distal myopathy with rimmed vacuoles: impaired O-glycan formation in muscular glycoproteins. AB - Distal myopathy with rimmed vacuoles (DMRV), is an autosomal recessive disorder with early adult onset, displays distal dominant muscular involvement and is characterized by the presence of numerous rimmed vacuoles in the affected muscle fibers. The pathophysiology of DMRV has not been clarified yet, although the responsible gene was identified as that encoding UDP-N-acetylglucosamine 2 epimerase/N-acetylmannosamine kinase involved in the biosynthesis of sialic acids. To identify defective carbohydrate moieties of muscular glycoproteins from DMRV patients, frozen skeletal muscle sections from seven patients with DMRV, as well as normal and pathological controls, were treated with or without sialidase or N-glycosidase F followed by lectin staining and lectin blotting analysis. The sialic acid contents of the O-glycans in the skeletal muscle specimens from the DMRV patients were also measured. We found that Arachis hypogaea agglutinin (PNA) lectin reacted strongly with sarcolemmal glycoproteins in the DMRV patients but not with those in control subjects. alpha-Dystroglycan from the DMRV patients strongly associated with PNA lectin, although that from controls did not. The sialic acid level of the O-glycans in the DMRV muscular glycoproteins with molecular weights of 30 to 200 kd was reduced to 60 to 80% of the control level. The results show that impaired sialyl O-glycan formation in muscular glycoproteins, including alpha-dystroglycan, occurs in DMRV. PMID- 15793293 TI - Interleukin-15 administration improves diaphragm muscle pathology and function in dystrophic mdx mice. AB - Interleukin (IL)-15, a cytokine expressed in skeletal muscle, has been shown to have muscle anabolic effects in vitro and to slow muscle wasting in rats with cancer cachexia. Whether IL-15 has therapeutic potential for diseases such as Duchenne muscular dystrophy (DMD) is unknown. We examined whether IL-15 administration could ameliorate the dystrophic pathology in the diaphragm muscle of the mdx mouse, an animal model for DMD. Four weeks of IL-15 treatment improved diaphragm strength, a highly significant finding because respiratory function is a mortality predictor in DMD. Enhanced diaphragm function was associated with increased muscle fiber cross-sectional area and decreased collagen infiltration. IL-15 administration was not associated with changes in T-cell populations or alterations in specific components of the ubiquitin proteasome pathway. To determine the effects of IL-15 on myofiber regeneration, muscles of IL-15-treated and untreated wild-type mice were injured myotoxically, and their functional recovery was assessed. IL-15 had a mild anabolic effect, increasing fiber cross sectional area after 2 and 6 days but not after 10 days. Our findings demonstrate that IL-15 administration improves the pathophysiology of dystrophic muscle and highlight a possible therapeutic role for IL-15 in the treatment of neuromuscular disorders especially in which muscle wasting is indicated. PMID- 15793294 TI - tPA receptors and the fibrinolytic response in multiple sclerosis lesions. AB - Axonal damage in multiple sclerosis (MS) lesions is associated with failure of fibrinolysis because of the inhibition of the plasminogen activator system. Plasma membrane receptors for tissue plasminogen activator (tPA) and plasminogen concentrate proteolytic activity on the cell surface and provide protection from inhibitors that in turn may locally enhance the fibrinolytic response. Therefore, we have investigated expression of two of these receptors in MS lesions, annexin II tetramer (AIIt) and low-density lipoprotein receptor-related protein (LRP). In acute MS lesions both AIIt and LRP were immunolocalized on macrophages and astrocytes while LRP was additionally found on neuronal cells in cortical gray matter. Western blot analysis confirmed a significant increase in AIIt in MS lesions and in a proportion of normal-appearing white matter samples, with a highly significant correlation between annexin II levels and factors associated with impeded fibrinolysis, such as plasminogen activator inhibitor-1. Immunoblotting analysis of plasmin(ogen) revealed increased levels of lysine plasminogen in samples expressing high AIIt protein levels. Our results suggest that limited availability of tPA in MS lesions because of formation of tPA plasminogen activator inhibitor-1 complexes reduces capability of tPA receptors to generate plasmin, which further diminishes fibrinolytic capacity in active MS lesions and possibly leads to axonal damage. PMID- 15793295 TI - Insulin-like growth factor II is involved in the proliferation control of medulloblastoma and its cerebellar precursor cells. AB - Medulloblastomas (MBs), the most frequent malignant brain tumors of childhood, presumably originate from cerebellar neural precursor cells. An essential fetal mitogen involved in the pathogenesis of different embryonal tumors is insulin like growth factor II (IGF-II). We screened human MB biopsies of the classic (CMB) and desmoplastic (DMB) variants for IGF2 transcripts of the four IGF2 promoters. We found IGF2 transcription from the imprinted promoter P3 to be significantly increased in the desmoplastic variant compared to the classic subgroup. This was not a result of loss of imprinting of IGF2 in desmoplastic tumors. We next examined the interaction of IGF-II and Sonic hedgehog (Shh), which serves as a critical mitogen for cerebellar granule cell precursors (GCPs) in the external granule cell layer from which DMBs are believed to originate. Mutations of genes encoding components of the Shh-Patched signaling pathway occur in approximately 50% of DMBs. To analyze the effects of IGF-II on Hedgehog signaling, we cultured murine GCP and human MB cells in the presence of Shh and Igf-II. In GCPs, a synergistic effect of Shh and Igf-II on proliferation and gli1 and cyclin D1 mRNA expression was found. Igf-II, but not Shh, induced phosphorylation of Akt and its downstream target Gsk-3beta. In six of nine human MB cell lines IGF-II displayed a growth-promoting effect that was mediated mainly through the IGF-I receptor. Together, our data point to an important role of IGF II for the proliferation control of both cerebellar neural precursors and MB cells. PMID- 15793296 TI - Microglia lacking E Prostanoid Receptor subtype 2 have enhanced Abeta phagocytosis yet lack Abeta-activated neurotoxicity. AB - Experimental therapies for Alzheimer's disease (AD) are focused on enhanced clearance of neurotoxic Abeta peptides from brain. Microglia can be neuroprotective by phagocytosing Abeta; however, this comes at the cost of activated innate immunity that causes paracrine damage to neurons. Here, we show that ablation of E prostanoid receptor subtype 2 (EP2) significantly increased microglial-mediated clearance of Abeta peptides from AD brain sections and enhanced microglial Abeta phagocytosis in cell culture. The enhanced phagocytosis was PKC-dependent and was associated with elevated microglial secretion of the chemoattractant chemokines, macrophage inflammatory protein-1alpha and macrophage chemoattractant protein-1. This suggested that microglial activation is negatively regulated by EP2 signaling through suppression of prophagocytic cytokine secretion. However, despite this enhancement of Abeta phagocytosis, lack of EP2 completely suppressed Abeta-activated microglia-mediated paracrine neurotoxicity. These data demonstrate that blockade of microglial EP2 is a highly desirable mechanism for AD therapy that can maximize neuroprotective actions while minimizing bystander damage to neurons. PMID- 15793297 TI - Matrix metalloproteinase activity and osteoclasts in experimental prostate cancer bone metastasis tissue. AB - Previously, we and others showed that broad spectrum pharmaceutical inhibition of matrix metalloproteinase (MMP) activity reduces intraosseous tumor burden and bone degradation in animal models of bone metastasis. Herein, we used specific assays to measure net enzymatic activities of individual MMPs during colonization of bone by prostate cancer cells. PC3 cells were injected into the marrow of human fetal femurs previously implanted in SCID mice. Net MMP-9 activity in bone tissues peaked 2 weeks after injection, coinciding with a wave of osteoclast recruitment. In contrast, MMP-2 and MT1-MMP activity did not change. In vitro, co culture of PC3 cells with bone tissue led to activation of pro-MMP-9 and increases in secreted net MMP-9 activity. Activation of pro-MMP-9 was prevented by metalloprotease inhibitors but not by inhibitors of other classes of proteases. Ribozyme suppression of MMP-9 expression in PC3 cells did not affect pro-MMP-9 activation or net MMP-9 activity and did not affect the phenotype of bone tumors. siRNA targeting of MMP-9 expression in preosteoclasts in vitro demonstrated that tumor-induced preosteoclast motility was dependent on MMP-9 expression. These data suggest that osteoclast-derived MMP-9 may represent a potential therapeutic target in bone metastasis and provide a rationale for the development of MMP-9-specific inhibitors. PMID- 15793298 TI - Chromatin organization measured by AluI restriction enzyme changes with malignancy and is regulated by the extracellular matrix and the cytoskeleton. AB - Given that expression of many genes changes when cells become malignant or are placed in different microenvironments, we asked whether these changes were accompanied by global reorganization of chromatin. We reasoned that sequestration or exposure of chromatin-sensitive sites to restriction enzymes could be used to detect this reorganization. We found that AluI-sensitive sites of nonmalignant cells were relatively more exposed compared to their malignant counterparts in cultured cells and human tumor samples. Changes in exposure and sequestration of AluI-sensitive sites in normal fibroblasts versus fibrosarcoma or those transfected with oncogenes, nonmalignant breast cells versus carcinomas and poorly metastatic versus highly invasive melanoma were shown to be independent of the cell cycle and may be influenced by proteins rich in disulfide bonds. Remarkably, regardless of degree of malignancy, AluI-sensitive sites became profoundly sequestered when cells were incubated with laminin, Matrigel, or a circular RGD peptide (RGD-C), but became exposed when cells were placed on collagen I or in serum-containing medium. Disruption of the actin cytoskeleton led to exposure, whereas disruption of microtubules or intermediate filaments exerted a sequestering effect. Thus, AluI-sensitive sites are more sequestered with increasing malignant behavior, but the sequestration and exposure of these sites is exquisitely sensitive to information conferred to the cell by molecules and biomechanical forces that regulate cellular and tissue architecture. PMID- 15793299 TI - Gene expression analysis of immune-mediated arrest of tumorigenesis in a transgenic mouse model of HER-2/neu-positive basal-like mammary carcinoma. AB - We previously showed that a vaccine combining interleukin 12 and allogeneic p185(neu)-positive mammary carcinoma cells completely prevented multifocal mammary carcinogenesis in HER-2/neu transgenic mice. To identify the molecular events responsible for effective tumor prevention and to define the tumor gene expression signature, we used microarrays to analyze the expression profile of mammary tissue of untreated transgenic mice and of vaccine-treated, tumor-free mice at different time points. Mammary tissue from vaccinated mice displayed a gene expression profile different from that of untreated, tumor-bearing mice but similar to that of normal/hyperplastic mammary gland. Comparison of treated and untreated mice at 15 weeks of age revealed up-regulation of genes encoding antibodies, chemokines, gamma-interferon-induced genes and inflammatory molecules, and down-regulation of early genes induced by tumor development. The gene expression signature of HER-2/neu-transformed tumor cells showed modulation of genes promoting proliferation, angiogenesis, migration, invasion, and metastasis and inhibiting apoptosis and immune response. Meta-analysis of microarray data on human breast cancer showed that the signature of tumors arising in murine HER-2/neu transgenic model correctly classified human HER-2/neu expressing tumors and normal breast tissue. Moreover murine and human HER-2/neu positive tumors share the signature of basal-like breast cancers. This gene expression analysis reveals the immune events associated with prevention of tumor development and shows that HER-2/neu transgenic mice represent a good model of a poor-prognosis group of human breast tumors. PMID- 15793300 TI - Analysis of interleukin-27 (EBI3/p28) expression in Epstein-Barr virus- and human T-cell leukemia virus type 1-associated lymphomas: heterogeneous expression of EBI3 subunit by tumoral cells. AB - Interleukin (IL)-27 is a novel heterodimeric cytokine of the IL-12 family that is composed of two subunits, Epstein-Barr virus (EBV)-induced gene 3 (EBI3) and p28. EBI3 is expressed at high levels in EBV-transformed B-cell lines and is induced in vitro by the EBV oncogene LMP1 in a nuclear factor (NF)-kappaB-dependent manner. We show here that EBI3 expression is up-regulated in human T-cell leukemia virus type 1 (HTLV-1)-infected cell lines and IL-2-dependent leukemic cells from adult T-cell leukemia/lymphoma (ATL) patients, compared to normal activated T cells. EBI3 expression was decreased in HTLV-1-transformed cells after treatment with the NF-kappaB inhibitor BAY11-7082 and was induced in Jurkat cells by expression of HTLV-1 wild-type Tax oncoprotein, but not by the Tax mutant M22, which is defective for NF-kappaB activation. In situ analysis of EBI3 and p28 expression in Hodgkin's lymphomas (HLs), in various EBV-associated lymphoproliferative disorders (LPDs) (including post-transplant LPDs and nasal type NK/T-cell lymphomas), and in ATL showed that EBI3 was expressed by neoplastic cells in all cases of HL and of LMP1-positive EBV-associated LPD, at variable levels in ATL cases, but rarely in control T-cell lymphomas. In contrast, in all lymphomas tested, no or few tumoral cells expressed p28. Consistent with these data, no significant p28 or IL-27 expression was detected in HL-derived cell lines, or in EBV- or HTLV-1-transformed cell lines. This selective overexpression of EBI3 by transformed cells suggests that EBI3 may play a role, independently from its association to p28, in regulating anti-viral or anti-tumoral immune responses. PMID- 15793301 TI - Genomic amplification of the human telomerase gene (TERC) in pap smears predicts the development of cervical cancer. AB - Invasive cervical carcinomas almost invariably carry extra copies of chromosome arm 3q, resulting in a gain of the human telomerase gene (TERC). This provided the rationale for the development of a multicolor fluorescence in situ hybridization (FISH) probe set as a diagnostic tool for the direct detection of TERC gains in Pap smears. We previously used this probe set to show that cervical intraepithelial neoplasia (CIN) 2 and CIN3 lesions could be distinguished from normal samples, atypical squamous cell of undetermined significance (ASCUS) and CIN1, with a sensitivity and specificity exceeding 90%, independent of the cytomorphological assessment. In the current study, we explored whether gain of 3q and amplification of TERC could predict progression from CIN1/CIN2 to CIN3 and invasive carcinoma. We applied our probe set to a series of 59 previously stained Pap smears for which repeat Pap smears and clinical follow-up were available. The samples included CIN1/CIN2 lesions that progressed to CIN3 (progressors), CIN1/CIN2 lesions that regressed spontaneously (regressors), and normal Pap smears from women who subsequently developed CIN3 or cervical cancer. Here, we show that progressors displayed a gain of 3q whereas none of the regressors showed this genetic aberration. These data suggest that 3q gain is required for the transition from CIN1/CIN2 to CIN3 and that it predicts progression. Of note, 3q gain was found in 33% of cytologically normal Pap smears from women who were diagnosed with CIN3 or invasive cervical carcinoma after a short latency. The sensitivity of our test for predicting progression from CIN1/CIN2 to CIN3 was 100% and the specificity, ie, the prediction of regression, was 70%. We conclude that the detection of 3q gain and amplification of TERC in routinely collected Pap smears can assist in identifying low-grade lesions with a high progression risk and in decreasing false-negative cytological screenings. PMID- 15793302 TI - Inactivation of p21WAF1/cip1 enhances intestinal tumor formation in Muc2-/- mice. AB - In the Apc1638(+/-) mouse model of intestinal tumorigenesis, targeted inactivation of the cyclin-dependent kinase inhibitor p21(WAF1/cip1) is highly effective in enhancing Apc-initiated tumor formation in the intestine. Because p21(WAF1/cip1) plays a critical role in regulating intestinal cell proliferation, maturation, and tumorigenesis, we examined whether its inactivation would enhance tumor formation in a different mouse model of colon cancer. Therefore, we mated p21(-/-) mice with mice carrying a genetic deficiency of the Muc2 gene, which encodes the major gastrointestinal mucin. Muc2(-/-) mice develop tumors in the small and large intestine and the rectum, but in contrast to tumors in Apc1638(+/ ) mice, this does not involve increased expression or nuclear localization of beta-catenin. We found that inactivation of p21(WAF1/cip1) significantly increased the frequency and size of intestinal tumors in Muc2 knockout mice and also led to development of more invasive adenocarcinomas. This enhanced tumorigenesis significantly decreased mouse life span. Further, inactivation of p21(WAF1/cip1) increased cell proliferation, decreased apoptosis, and decreased intestinal trefoil factor expression in the mucosa of both the small and large intestine. Surprisingly, reduced expression of p27(kip1) was also observed in the Muc2(-/-), p21(+/-), and p21(-/-) mice. In contrast, the expression of c-myc was significantly elevated. Thus, p21 modulates the formation of tumors whose initiation does (Apc) or does not (Muc2) involve altered beta-catenin-Tcf4 signaling, but which may converge on common elements downstream of this signaling pathway. PMID- 15793303 TI - Receptor protein tyrosine phosphatase micro regulates the paracellular pathway in human lung microvascular endothelia. AB - The pulmonary vascular endothelial paracellular pathway and zonula adherens (ZA) integrity are regulated, in part, through protein tyrosine phosphorylation. ZA associated protein tyrosine phosphatase (PTP)s are thought to counterregulate tyrosine phosphorylation events within the ZA multiprotein complex. One such receptor PTP, PTPmu, is highly expressed in lung tissue and is almost exclusively restricted to the endothelium. We therefore studied whether PTPmu, in pulmonary vascular endothelia, associates with and/or regulates both the tyrosine phosphorylation state of vascular endothelial (VE)-cadherin and the paracellular pathway. PTPmu was expressed in postconfluent human pulmonary artery and lung microvascular endothelial cells (ECs) where it was almost exclusively restricted to EC-EC boundaries. In human lung microvascular ECs, knockdown of PTPmu through RNA interference dramatically impaired barrier function. In immortalized human microvascular ECs, overexpression of wild-type PTPmu enhanced barrier function. PTPmu-VE-cadherin interactions were demonstrated through reciprocal co immunoprecipitation assays and co-localization with double-label fluorescence microscopy. When glutathione S-transferase-PTPmu was incubated with purified recombinant VE-cadherin, and when glutathione S-transferase-VE-cadherin was incubated with purified recombinant PTPmu, PTPmu directly bound to VE-cadherin. Overexpression of wild-type PTPmu decreased tyrosine phosphorylation of VE cadherin. Therefore, PTPmu is expressed in human pulmonary vascular endothelia where it directly binds to VE-cadherin and regulates both the tyrosine phosphorylation state of VE-cadherin and barrier integrity. PMID- 15793304 TI - Angiogenetic signaling through hypoxia: HMGB1: an angiogenetic switch molecule. AB - The initiation of angiogenesis, called the angiogenetic switch, is a crucial early step in tumor progression and propagation, ensuring an adequate oxygen supply. The rapid growth of tumors is accompanied by a reduced microvessel density, resulting in chronic hypoxia that often leads to necrotic areas within the tumor. These hypoxic and necrotic regions exhibit increased expression of angiogenetic growth factors, eg, vascular endothelial growth factor, and may also attract macrophages, which are known to produce a number of potent angiogenetic cytokines and growth factors. A group of molecules that may act as mediators of angiogenesis are the so-called high-mobility group proteins. Recent studies showed that HMGB1, known as an architectural chromatin-binding protein, can be extracellularly released by passive diffusion from necrotic cells and activated macrophages. To examine the angiogenetic effects of HMGB1 on endothelial cells an in vitro spheroid model was used. The results of the endothelial-sprouting assay clearly show that exogenous HMGB1 induced endothelial cell migration and sprouting in vitro in a dose-dependent manner. Thus, this is the first report showing strong evidence for HMGB1-induced sprouting of endothelial cells. PMID- 15793305 TI - Macrophage conditioned medium induces the expression of C-reactive protein in human aortic endothelial cells: potential for paracrine/autocrine effects. AB - C-reactive protein (CRP) is a risk marker for cardiovascular events in apparently healthy persons. Cogent data show that, aside from the liver, CRP is produced in atherosclerotic lesions, kidney, neurons, and alveolar macrophages. Because several proatherogenic effects of CRP have been documented in endothelial cells, we examined human aortic endothelial cells (HAEC) for CRP production. We detected the presence of CRP mRNA by RT-PCR and in situ hybridization, intracellular protein by Western blot and secreted protein by ELISA. Coincubation with the cytokines interleukin (IL)-1, IL-6, and tumor necrosis factor alone and in combination showed that the most potent agonist for CRP production from HAEC is the combination of IL-1 and IL-6 (P < 0.05). To mimic the in vivo situation, we examined whether vascular smooth muscle cell (VSMC) and/or macrophage conditioned media (MCM) could augment CRP production by HAEC. While VSMC-conditioned media had no effect, incubation with MCM resulted in a significant twofold increase in the synthesis of both intracellular and secreted CRP (P < 0.05). The effect of MCM could be reversed by inhibiting both IL-1 and IL-6. Thus, stimulated synthesis and secretion of CRP by cells in the atherosclerotic lesion by paracrine/autocrine loops could result in local concentrations of CRP far in excess of plasma concentrations and could contribute to proinflammatory, proatherogenic effects. PMID- 15793307 TI - Visualization of single Escherichia coli FtsZ filament dynamics with atomic force microscopy. AB - FtsZ, the prokaryotic homologue of tubulin, is an essential cell division protein. In the cell, it localizes at the center, forming a ring that constricts during division. In vitro, it binds and hydrolyzes GTP and polymerizes in a GTP dependent manner. We have used atomic force microscopy to study the structure and dynamics of FtsZ polymer assembly on a mica surface under buffer solution. The polymers were highly dynamic and flexible, and they continuously rearranged over the surface. End-to-end joining of filaments and depolymerization from internal zones were observed, suggesting that fragmentation and reannealing may contribute significantly to the dynamics of FtsZ assembly. The shape evolution of the restructured polymers manifested a strong inherent tendency to curve. Polymers formed in the presence of non-hydrolyzable nucleotide analogues or in the presence of GDP and AlF(3) were structurally similar but showed a slower dynamic behavior. These results provide experimental evidence supporting the model of single-strand polymerization plus cyclization recently proposed to explain the hydrodynamic behavior of the polymers in solution. PMID- 15793306 TI - Caveolae participate in tumor necrosis factor receptor 1 signaling and internalization in a human endothelial cell line. AB - Caveolae are abundant in endothelial cells (ECs) in situ but markedly diminished in cultured cells, making it difficult to assess their role in cytokine signaling. We report here that the human EC line EA.hy926 retains an abundant caveolar system in culture. Tumor necrosis factor (TNF) receptor 1 (TNFR1/CD120a) was enriched in caveolae and co-immunoprecipitated with caveolin-1 from caveolae isolated from these cells. To further investigate the role(s) of caveolae in TNF signaling in ECs, cells were treated with methyl-beta-cyclodextrin to disrupt caveolae. Methyl-beta-cyclodextrin did not alter total cell surface expression of TNFR1 or TNF-induced degradation of IkappaBalpha, a measure of nuclear factor kappaB activation, but it did inhibit TNF-induced phosphorylation of Akt, a measure of phosphatidylinositol-3 kinase activation. Serum-induced phosphorylation of AKT was unaffected. Treatment with TNF induced disappearance of TNFR1 from caveolae and dissociation from caveolin-1 within 5 minutes. In contrast to transferrin receptor, internalized TNFR1 did not co-localize with clathrin, except possibly in the Golgi, at any time point examined. By 60 minutes of treatment with TNF, TNFR1 appeared in endosomes. We conclude that caveolae function in ECs to allow TNFR1 to activate phosphatidylinositol-3 kinase and Akt, perhaps through receptor cross talk, and that ligand-induced internalization and trafficking of TNFR1 to endosomes may originate directly from this compartment. PMID- 15793309 TI - Local atrial natriuretic peptide signaling prevents hypertensive cardiac hypertrophy in endothelial nitric-oxide synthase-deficient mice. AB - The crucial functions of atrial natriuretic peptide (ANP) and endothelial nitric oxide/NO in the regulation of arterial blood pressure have been emphasized by the hypertensive phenotype of mice with systemic inactivation of either the guanylyl cyclase-A receptor for ANP (GC-A-/-) or endothelial nitric-oxide synthase (eNOS-/ ). Intriguingly, similar levels of arterial hypertension are accompanied by marked cardiac hypertrophy in GC-A-/-, but not in eNOS-/-, mice, suggesting that changes in local pathways regulating cardiac growth accelerate cardiac hypertrophy in the former and protect the heart of the latter. Our recent observations in mice with conditional, cardiomyocyte-restricted GC-A deletion demonstrated that ANP locally inhibits cardiomyocyte growth. Abolition of these local, protective effects may enhance the cardiac hypertrophic response of GC-A-/ mice to persistent increases in hemodynamic load. Notably, eNOS-/- mice exhibit markedly increased cardiac ANP levels, suggesting that increased activation of cardiac GC-A can prevent hypertensive heart disease. To test this hypothesis, we generated mice with systemic inactivation of eNOS and cardiomyocyte-restricted deletion of GC-A by crossing eNOS-/- and cardiomyocyte-restricted GC-A-deficient mice. Cardiac deletion of GC-A did not affect arterial hypertension but significantly exacerbated cardiac hypertrophy and fibrosis in eNOS-/- mice. This was accompanied by marked cardiac activation of both the mitogen-activated protein kinase (MAPK) ERK 1/2 and the phosphatase calcineurin. Our observations suggest that local ANP/GC-A/cyclic GMP signaling counter-regulates MAPK/ERK- and calcineurin/nuclear factor of activated T cells-dependent pathways of cardiac myocyte growth in hypertensive eNOS-/- mice. PMID- 15793308 TI - Hedgehog signaling induces cardiomyogenesis in P19 cells. AB - Sonic Hedgehog (Shh) is a critical signaling factor for a variety of developmental pathways during embryogenesis, including the specification of left right asymmetry in the heart. Mice that lack Hedgehog signaling show a delay in the induction of cardiomyogenesis, as indicated by a delayed expression of Nkx2 5. To further examine a role for Shh in cardiomyogenesis, clonal populations of P19 cells that stably express Shh, termed P19(Shh) cells, were isolated. In monolayer P19(Shh) cultures the Shh pathway was functional as shown by the up regulation of Ptc1 and Gli1 expression, but no cardiac muscle markers were activated. However, Shh expression induced cardiomyogenesis following cellular aggregation, resulting in the expression of factors expressed in cardiac muscle including GATA-4, MEF2C, and Nkx2-5. Furthermore, aggregated P19 cell lines expressing Gli2 or Meox1 also up-regulated the expression of cardiac muscle factors, leading to cardiomyogenesis. Meox1 up-regulated the expression of Gli1 and Gli2 and, thus, can modify the Shh signaling pathway. Finally, Shh, Gli2, and Meox1 all up-regulated BMP-4 expression, implying that activation of the Hedgehog pathway can regulate bone morphogenetic protein signals. Taken together, we propose a model in which Shh, functioning via Gli1/2, can specify mesodermal cells into the cardiac muscle lineage. PMID- 15793310 TI - Lipopolysaccharide improves cardiomyocyte survival and function after serum deprivation. AB - Toll-like receptor-4 (TLR4) and its signaling molecule interleukin-1 receptor associated kinase (IRAK-1) play an important role in host defense and tissue inflammation. Intriguingly, systemic administration of lipopolysaccharide (LPS), the agonist for TLR4, confers a cardio-protective effect against ischemic injury. However, the mechanisms leading to the cardiac protection remain largely unknown. The present study was designed to investigate the role of TLR4 activation by LPS in protecting cardiomyocytes (CM) against apoptosis in an in vitro model of ischemia and to explore the downstream mechanisms leading to the protective effect. Incubation with LPS led to activation of IRAK-1 and protected CMs against serum deprivation (SD)-induced apoptosis as demonstrated by DNA laddering, histone-DNA fragment enzyme-linked immunosorbent assay, and activation of caspase 3. Phosphatidylinositol 3-kinase/Akt, extracellular signal-regulated kinase 1/2, and IkappaB kinase beta appear to contribute to the anti-apoptotic effect of LPS since the specific inhibitors, wortmannin, PD98059, and dominant negative IKKbeta transgene expression reversed the LPS effect. To assess whether LPS improves CM function, we examined intracellular Ca(2+) transients and cell shortening in single adult rat CMs. SD for 6 h dramatically inhibited Ca(2+) transients and CM contractility. LPS at 500 ng/ml significantly improved the [Ca(2+)](i) transients and enhanced contractility in control CMs as well as in CMs subjected to SD. Importantly, transient ischemia led to rapid activation of IRAK-1 in cultured CMs and in adult rat myocardium. Adenovirus-mediated transgene expression of IRAK-1 but not its kinase-deficient mutant IRAK-1(K239S) protected CMs against SD induced apoptosis. Taken together, these data suggest an important role of TLR4 signaling via IRAK-1 in protecting against SD-induced apoptosis. PMID- 15793311 TI - Divergent distribution in vascular and avascular mammalian retinae links neuroglobin to cellular respiration. AB - The visual function of the vertebrate retina relies on sufficient supply with oxygen. Neuroglobin is a respiratory protein thought to play an essential role in oxygen homeostasis of neuronal cells. For further understanding of its function, we compared the distribution of neuroglobin and mitochondria in both vascular and avascular mammalian retinae. In the vascular retinae of mouse and rat, oxygen is supplied by the outer choroidal, deep retinal, and inner capillaries. We show that in this type of retina, mitochondria are concentrated in the inner segments of photoreceptor cells, the outer and the inner plexiform layers, and the ganglion cell layer. These are the same regions in which oxygen consumption takes place and in which neuroglobin is present at high levels. In the avascular retina of guinea pig the deep retinal and inner capillaries are absent. Therefore, only the inner segments of the photoreceptors adjacent to choroidal capillaries display an oxidative metabolism. We demonstrate that in the retina of guinea pigs both neuroglobin and mitochondria are restricted to this layer. Our results clearly demonstrate an association of neuroglobin and mitochondria, thus supporting the hypothesis that neuroglobin is a respiratory protein that supplies oxygen to the respiratory chain. PMID- 15793312 TI - Clostridium difficile: an update. AB - Clostridium difficile causes a spectrum of diarrheal illness with the potential for major medical consequences. Although most cases respond quickly to treatment, C. difficile colitis may be severe and life threatening. Recurrent disease represents a particularly challenging problem. PMID- 15793313 TI - Behavioral counseling interventions for primary prevention of coronary heart disease in individuals with type-2 diabetes. AB - Coronary heart disease (CHD) is the leading cause of death in diabetes. CHD risk can be altered by lifestyle modification. Behavioral counseling interventions are effective at changing behavior. This article discusses effective counseling techniques in primary care for people with diabetes. PMID- 15793314 TI - Thyroid hormones therapy: a review of their effects in the treatment of psychiatric and medical conditions. AB - Although thyroid hormones are indicated for hypothyroidism, their therapeutic benefit in treating other medical and psychiatric conditions has not been adequately studied. We review the use of thyroid hormones in treating psychiatric and medical conditions in the euthyroid patients. PMID- 15793315 TI - A woman's body and her vascular system are unique. How should we care for them? AB - Women must now gain the phenomenal care justified by new biologic evidence and a comprehensive understanding of fundamental differences from men-that go well beyond secondary sexual characteristics. PMID- 15793316 TI - Recurrent Clostridium difficile colitis. AB - Clostridium difficile is the most common cause of nosocomial infectious diarrhea that is usually treated adequately with standard treatment of metronidazole or vancomycin. Relapse or recurrent infection can occur in certain patients and this can be very difficult to treat. PMID- 15793317 TI - Current concepts and future directions in the battle against obesity. AB - Obesity is a significant unsolved public health problem. Future treatments of obesity will involve application of neuroendocrine concepts that are reviewed in this paper. PMID- 15793318 TI - Acute pelvic pain: evaluation and management. AB - Acute pelvic pain in women is often a diagnostic dilemma. Obstetrical, gynecological, urological or gastrointestinal causes must be considered. Stabilization, immediate therapy and early consultation are often indicated. If no etiology is found, conservative management with frequent re-evaluation is adequate. PMID- 15793320 TI - Musculoskeletal examination: a complete review. AB - The musculoskeletal system provides the most obvious visual clues to a person's health. Problems in the musculoskeletal system can be intrinsic muscle problems or referred from organs. The history and physical examination, which are important in assessing the health of this system, are reviewed. Dynamic examination for assessing the functioning of the system are needed and are outlined. The aging of muscles and how it affects muscle function is discussed. PMID- 15793319 TI - Diagnosing occupational lung disease: a practical guide to the occupational pulmonary history for the primary care practitioner. AB - A well-executed occupational pulmonary history should be part of the evaluation of workers presenting with respiratory illnesses or symptoms. In this article, we review the scope of occupational lung disease and detail the essential elements of the occupational pulmonary history. PMID- 15793321 TI - Sleep-disordered breathing: implications for the pathophysiology and management of cardiovascular disease. AB - Sleep apnea is increasingly associated with risk of cardiovascular disease, including arrhythmia, heart failure, stroke, ischemic heart disease, and hypertension. Diagnosis and treatment of sleep apnea and the implications for cardiovascular disease are discussed. PMID- 15793322 TI - Chronic pelvic pain in women: evaluation and management in primary care. AB - Chronic pelvic pain (CPP) in women is a common disorder, affecting as many as 15% of adult women, and often provides both a diagnostic and therapeutic challenge. Pain in CPP may originate directly from pelvic organs, or may be referred from more distant tissue sites. A comprehensive medical history and physical examination should include special attention to gynecological, urological, gastrointestinal, psychiatric, myofascial, and neuromuscular systems. The effective management of CPP may involve comprehensive evaluations by specialists, psychologists, and multiple office visits. Physicians should address CPP as a chronic disease. Combining lifestyle modification with other traditional treatments produces better outcomes. Laboratory tests, transvaginal ultrasound, and laparoscopy may identify serious disease or provide significant reassurance to patient. Specific surgical procedures for various conditions and pain relief measures are beneficial in selected patients. A sensitive physician who is willing to spend adequate time and coordinate care with specialists can markedly diminish the suffering of these patients. PMID- 15793323 TI - Practical approach to the management of chronic pain. AB - Chronic pain is a common and disabling complaint in most primary care practices. This article offers an explanation for the physiological basis of persistent chronic pain, provides practical guidelines to help distinguish among common pain syndromes, and recommends treatment strategies. PMID- 15793324 TI - The comprehensive therapy of diabetic foot ulcers. AB - Foot wounds are a common complication of diabetes mellitus, and a significant number of them lead to amputation. Fortunately, the majority of diabetic foot wounds are preventable. In this article, we review the different factors leading to ulceration and outline the essentials of a thorough evaluation. Finally, we present the various management strategies available to the treating physician. PMID- 15793325 TI - Disorders of flushing. AB - Disorders of flushing encompass a broad spectrum of diverse acquired and inherited conditions. Chemical mediators involved in the flushing response are incompletely understood. Flushing episodes rarely can be associated with significant morbidity and mortality. The goal of the physician is to separate benign from potentially life-threatening conditions. Accurate diagnosis requires a thorough history and physical examination emphasizing the age of the patient, temporal association of flushing with occupation, environmental, stress, food, or drug exposure, and the duration of the episode. In some cases, despite a thorough evaluation, the etiology for flushing remains unknown. Understanding the distinct mechanisms that lead to flushing helps provide a rational approach to treatment. PMID- 15793327 TI - Cultural influences on pain management in the elderly. AB - Cultural influences are often more important in the elderly and help explain the influence of nonbiological factors underlying pain appraisal, expression, and folk treatments. Five main psychological theories that underlie these cultural influences help direct supportive interventions. PMID- 15793326 TI - Protection of human subjects in clinical research: the pitfalls in clinical research. AB - Clinical trials are an important part of medical research and must be done with good conduct and intention. Using a suitable approach is part of research practice, which favors protection of research subjects. Good Clinical Practice is being promulgated as a universal ethical approach toward protection of the human subject's rights. Appropriate scientific design, good performance, better analyses, informed consent, and credible scientific data with the confidentiality of the research subject all contribute to Good Clinical Practices. The first and possibly the most important step toward protection of the research subject is to appropriately obtain an informed consent, which identifies respect, beneficence, and justice for the enrolled research subjects. PMID- 15793328 TI - Women and alcohol. AB - Primary care providers are uniquely positioned to educate women about alcohol use and misuse. Early identification and intervention can significantly limit the adverse consequences of alcohol abuse. Physicians must recognize the risk factors, presentation, and treatment issues relevant to women. PMID- 15793329 TI - From the pen to the patient: minimising medication errors. PMID- 15793331 TI - Irrational drug use in India: a prescription survey from Goa. AB - BACKGROUND: There is concern regarding the irrational production, prescription and use of drugs in India. This study aimed to describe the quality of prescriptions by medical practitioners, including both the layout of the prescription and the type and number of drugs prescribed. MATERIALS AND METHODS: A survey of all prescriptions dispensed at a busy pharmacy in the state of Goa, India, was carried out over a consecutive seven-day period. Each prescription was rated on the basis of a priori and pilot-tested variable list. The prescriptions by private practitioners were compared with those from practitioners in the public healthcare system. RESULTS: Nine hundred and ninety prescriptions were collected. The majority (83.9%) were from private practitioners. The quality of the layout of the prescriptions was unsatisfactory: information to identify the practitioner was incomplete in more than a third of the prescriptions and information to identify the patient was incomplete in more than half. Clarity of written instructions on how to take the medicines was unsatisfactory in the majority of prescriptions. Polypharmacy was the norm, with more than half (52.7%) the prescriptions containing at least 3 medicines. Forty per cent of prescriptions included a vitamin or tonic preparation and a quarter of the prescriptions included an antibiotic and an analgesic. Over 90% of prescriptions contained only branded medicines. Private practitioners prescribed significantly greater number of medicines and were more likely to prescribe vitamins and antibiotics, and branded medicines. DISCUSSION: This study confirms that the quality of prescriptions, both in terms of layout and the content of the drugs prescribed, is inadequate. There is a need to standardize the format of prescriptions in India so that all essential information is included. There is a need to strengthen an independent mechanism for continuing professional development of practitioners to ensure that patients are always given evidence based, cost-effective treatments. PMID- 15793332 TI - Confusing brand names: nightmare of medical profession. AB - OBJECTIVE: India has more than 20,000 registered pharmaceutical manufacturers. Consequently, there is a flood of brand names to choose from. We conducted this study to analyse and sort out the multitudinous brand names thronging the Indian market, and identified those that could create a possible confusion. MATERIALS AND METHODS: Recent issues of drug formularies like Indian Drug Review, Drug Index, and Monthly Index of Medical Specialities-India were checked and all the brand names given were included. Some other brand names that are available with the pharmacists but are not included in these indexes were also included in the study for analysis. OBSERVATIONS: Potentially confusing brand names were sorted out and categorised according to the severity of damage they can cause if misinterpreted by the pharmacist or the patient. Subgroups were made according to the brand name, the generic name, and the manufacturers of the drug. CONCLUSION: Several brand names are strikingly identical, similar looking (orthographic), or similar sounding (phonological). Preventing this possible confusion is not the work of any one person involved. We describe the role of prescribing doctors, dispensing pharmacists, consumer patients, and the manufacturing companies to prevent "wrong prescribing" due to similarities in brand names. PMID- 15793333 TI - Conversion from laparoscopic to open cholecystectomy: multivariate analysis of preoperative risk factors. AB - BACKGROUND: Laparoscopic cholecystectomy has become the gold standard in the treatment of symptomatic cholelithiasis. Some patients require conversion to open surgery and several preoperative variables have been identified as risk factors that are helpful in predicting the probability of conversion. However, there is a need to devise a risk-scoring system based on the identified risk factors to (a) predict the risk of conversion preoperatively for selected patients, (b) prepare the patient psychologically, (c) arrange operating schedules accordingly, and (d) minimize the procedure-related cost and help overcome financial constraints, which is a significant problem in developing countries. AIM: This study was aimed to evaluate preoperative risk factors for conversion from laparoscopic to open cholecystectomy in our setting. SETTINGS AND DESIGNS: A case control study of patients who underwent laparoscopic surgery from January 1997 to December 2001 was conducted at the Aga Khan University Hospital, Karachi, Pakistan. MATERIALS AND METHODS: All those patients who were converted to open surgery (n = 73) were enrolled as cases. Two controls who had successful laparoscopic surgery (n = 146) were matched with each case for operating surgeon and closest date of surgery. STATISTICAL ANALYSIS USED: Descriptive statistics were computed and, univariate and multivariate analysis was done through multiple logistic regression. RESULTS: The final multivariate model identified two risk factors for conversion: ultrasonographic signs of inflammation (adjusted odds ratio [aOR] = 8.5; 95% confidence interval [CI]: 3.3, 21.9) and age > 60 years (aOR = 8.1; 95% CI: 2.9, 22.2) after adjusting for physical signs, alkaline phosphatase and BMI levels. CONCLUSION: Preoperative risk factors evaluated by the present study confirm the likelihood of conversion. Recognition of these factors is important for understanding the characteristics of patients at a higher risk of conversion. PMID- 15793334 TI - Percutaneous endoscopic gastrostomy: 30-day mortality trends and risk factors. AB - BACKGROUND AND AIMS: 30-day Percutaneous endoscopic gastrostomy (PEG) mortality of 8% (1992). Recent concerns suggest that mortality may have increased, prompting a comparison of current practice with that reported earlier. MATERIALS AND METHODS: Data regarding PEG insertion with relation to case mix, complications, 30-day mortality and associated risk factors, in 2002, in a British University Hospital was compared with that in 1992. Logistic regression analysis was used to determine factors independently predictive of 30-day mortality. RESULTS: In 2002, 112 patients (70% males, mean age 67.5 years; 1992: 63.6 years) underwent PEG. The 30-day mortality increased significantly from 8% (1992) to 22% (2002), P= 0.03. During this time, PEG insertion rate increased ten fold, however, procedure-related mortality decreased from 2% to nil. In terms of percentage, the indications for PEG in 1992 and 2002 respectively were: cerebrovascular disease (33/25), head and neck tumours (16/24), motor neuron disease (27/11, P= 0.01). The proportion of PEGs for non-evidence-based indications increased from 16% in 1992 to 31% in 2002, P= 0.048. The number of PEGs placed radiologically increased (0/17, P= 0.02). Radiological patients received less antibiotic prophylaxis (P< 0.001) and had more PEG site infections than standard placement, P= 0.04. Multivariate analysis identified nil by mouth > or = 7 days or 11.4 (CI 3.2-41.7), albumin< or = 30 g/L or 12 (2.2-66.7) and > 1 cardiac factor or 5.1 (1.02-25.6) as independent predictors of 30-day mortality. CONCLUSIONS: The ten-fold rise in the PEG insertion rate has been accompanied by a three-fold rise in 30-day mortality. This may reflect a lowered threshold of PEG insertion. The risk factors identified may help decision-making in cases where the risk-benefit relationship is not clear-cut. PMID- 15793335 TI - Sino-nasal epithelial tumours: a pathological study of 69 cases. AB - BACKGROUND: Epithelial neoplasms are uncommon lesions affecting the sino-nasal tract. There are hardly any reports in the Indian literature. AIM: To study the incidence, mode of presentation and histological types of sino-nasal epithelial tumours in the surgical pathology material. SETTING AND DESIGN: Retrospective retrieval of all sino-nasal tumours and analysis of epithelial tumours. MATERIALS AND METHODS: All sino-nasal epithelial tumours, biopsied or surgically excised over a period of ten years, were studied. The tumours were classified as benign or malignant. The histology was correlated with the clinical presentation and investigations. RESULTS: In ten years, there were 120 sino-nasal tumours representing 0.14% of all the surgical specimens received. Sixty-nine epithelial tumours (59.2%) outnumbered the non-epithelial tumours and were diagnosed on the basis of histopathology. Twenty were benign and 49 malignant; occurring predominantly in males. Benign lesions included four squamous papillomas and 16 inverted papillomas, with recurrence in three inverted papillomas (21%). Squamous cell carcinomas were the commonest among malignant tumours and four of these were associated with inverted or cylindrical cell papilloma. The second most frequent malignant tumour was adenoid cystic carcinoma with eight cases. Other rare types included the variants of squamous carcinoma, adenocarcinomas of the non-enteric type, muco-epidermoid carcinoma and undifferentiated carcinomas. CONCLUSION: Sino nasal epithelial tumours are rare lesions, with male preponderance. Inverted papillomas and squamous cell carcinomas are the most frequent neoplasms. PMID- 15793336 TI - Granular cell tumour of the ampulla of Vater. AB - Granular cell tumour (GCT) is a rare soft tissue neoplasm commonly encountered in the head and neck region, skin and subcutaneous tissue. GCT of the biliary system is most commonly reported in African-American females and usually presents as abdominal pain and obstructive jaundice. The neoplasm constitutes less than 10% of all benign tumours of the extra-hepatic biliary tree. No case of GCT involving the ampulla of Vater has been reported in the literature to date. We report a case of benign GCT involving the ampullary region in a 44-year-old Ethiopian male. Preoperative diagnosis was available on ERCP and deep biopsy. The patient was managed by debulking resection and biliary-enteric bypass and is symptom-free with no evidence of tumour progression after a follow-up of one year. PMID- 15793337 TI - Nasal dermoid with intracranial extension. AB - Nasal dermoids are rare congenital anomalies, which usually present in early childhood as a midline nasal mass that requires surgical management. It is important to rule out any intracranial extension before subjecting the patient to surgery. We describe a case of nasal dermoid with intracranial extension. The embryology of the formation of the nasal dermoid, cyst and sinus, with the role of CT scan and MRI in diagnosing intracranial extension, and its management are discussed. PMID- 15793338 TI - Chest wall metastases from unknown primary hepatocellular carcinoma. AB - Metastases of hepatocellular carcinoma (HCC) to the bones are common but bone metastases of hepatocellular carcinoma in the presence of a normal liver are an uncommon entity. A 50-year-old male patient presented with a rapidly growing tumour on the sternum. Biopsy of the lesion showed metastatic sternal tumour from a primary hepatocellular carcinoma. Radiological evaluation however, failed to detect a primary lesion in the liver. Bone metastases of hepatocellular carcinoma localized to the chest wall in the presence of a normal liver are scarcely reported as anecdotal case reports in the literature. PMID- 15793339 TI - Primary biliary cirrhosis complicated by transverse myelitis in a patient without Sjogren's syndrome. AB - Transverse myelitis is an acute inflammatory process, affecting one or more segments of the spinal cord. Its association with primary biliary cirrhosis has been documented in only four cases--all along with Sjogren's syndrome. Herein, we report for the first time, a patient who developed recurrent acute transverse myelitis in association with primary biliary cirrhosis without any clinical or histological indication of Sjogren's syndrome. A 42-year-old woman with primary biliary cirrhosis developed acute onset quadriparesis and urinary retention. Diagnostic evaluation excluded the presence of Sjogren's syndrome, other autoimmune syndromes, infections and multiple sclerosis. Magnetic resonance imaging of the spinal cord disclosed signal intensity abnormalities from C1 to T2 after gadolinium enhancement. As diagnosis of acute transverse myelitis was prominent, the patient was treated with intravenous methylprednisolone. The patient had a fair outcome despite an early recurrence of the symptoms after treatment withdrawal. PMID- 15793340 TI - Extra-medullary haematopoiesis in beta-thalassaemia. PMID- 15793341 TI - Solitary nevus lipomatosus cutaneous superficialis. PMID- 15793342 TI - A giant left atrium. PMID- 15793343 TI - An elderly lady in shock. PMID- 15793344 TI - Current perspectives on the management of gastroparesis. AB - Gastroparesis is a condition of abnormal gastric motility characterised by delayed gastric emptying in the absence of mechanical outlet obstruction. It is seen commonly in people with diabetes but is idiopathic in a third of patients. Symptoms include nausea and vomiting, post-prandial fullness and early satiety, and abdominal bloating and discomfort. Investigations fall into three categories: gastric emptying studies, intraluminal pressure measurements and recording of gastric myoelectrical activity. Nuclear scintigraphy is considered the gold standard for diagnosing and quantifying delayed gastric emptying. Treatment options include diet and behavioural changes, prokinetic drugs and surgical interventions. New advances in drug therapy and gastric electrical stimulation techniques hold considerable promise. PMID- 15793345 TI - Crossing to safety: transforming healthcare organizations for patient safety. AB - The current healthcare system is not designed to ensure better patient safety. In addition, healthcare is simultaneously becoming increasingly complex and increasingly fragmented. Medical knowledge and technology are expanding at an incredible rate, making it difficult for the healthcare providers to keep pace with advancing knowledge. Patients' needs are changing too: shifting from the diagnosis and treatment of a single, acute problem to the long-term management of multiple, interrelated chronic conditions. Our systems of care are not keeping up with these changes and, consequently, patients are experiencing unnecessary risk. Improving patient safety requires a transformation in how we currently care for patients. Healthcare organizations must adopt a new paradigm of care that holds patient safety as a core value and practice. To achieve this aim, healthcare organizations should build and maintain a culture of patient safety, provide leadership for patient safety that establishes a blame-free environment, proactively survey and monitor for adverse events, continually engineer patient safety into healthcare processes, and provide information and communication technologies to support patient safety. PMID- 15793346 TI - Insulin glargine: a long acting insulin analog. AB - The main goal of treatment of diabetes mellitus (DM) is to maintain long term near normoglycemia. Insulin therapy plays a pivotal role in the management of DM. Most insulin preparations and insulin delivery systems, do not mimic the physiological insulin secretion in the body, leading to impaired metabolic control and increased hypoglycemic attacks. Insulin glargine is newer, long acting insulin analog with duration of action of 24 hr. It practically does not show any peak over its duration of action. In various clinical trials, it has shown comparable/better efficacy than the currently used insulin replacement therapies with no increased side effects. In the current scenario, though it is difficult to achieve an ideal replacement therapy, insulin glargine is definitely a positive step in that direction. PMID- 15793347 TI - Bilateral post traumatic anterior shoulder dislocation. PMID- 15793348 TI - Vascular complication following lipid free propofol injection. PMID- 15793349 TI - Ileal metastases from oesophageal carcinoma causing intestinal obstruction. PMID- 15793350 TI - Severe autonomic dysfunction as a presenting feature of Wilson's disease. PMID- 15793351 TI - Yet another cause for drug-induced pulmonary fibrosis. PMID- 15793352 TI - Parasitic infections in HIV-infected patients who visited Seoul National University Hospital during the period 1995-2003. AB - The prevalence of parasitic infections was investigated in human immunodeficiency virus (HIV)-infected patients (n = 105) who visited Seoul National University Hospital, Seoul, Korea, during the period from 1995 to 2003. Fecal samples were collected from 67 patients for intestinal parasite examinations, and sputum or bronchoalveolar lavage samples from 60 patients for examination of Pneumocystis carinii. Both samples were obtained from 22 patients. Thirty-three (31.4%) of the 105 were found to have parasitic infections; Cryptosporidium parvum (10.5%; 7/67), Isospora belli (7.5%; 5/67), Clonorchis sinensis (3.0%; 2/67), Giardia lamblia (1.5%; 1/67), Gymnophalloides seoi (1.5%; 1/67), and Pneumocystis carinii (28.3%; 17/60). The hospital records of the 11 intestinal parasite-infected patients showed that all suffered from diarrhea. This study shows that parasitic infections are important clinical complications in HIV-infected patients in the Republic of Korea. PMID- 15793353 TI - Evaluation of taxonomic validity of four species of Acanthamoeba: A. divionensis, A. paradivionensis, A. mauritaniensis, and A. rhysodes, inferred from molecular analyses. AB - The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of 0.2-1.1% with each other, but more than 2% difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group II on the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes. PMID- 15793354 TI - Diagnostic value of a dot immunobinding assay for human pulmonary hydatidosis. AB - The diagnosis of human hydatidosis is primarily made using radiological and serological methods. Radiological methods are generally of low specificity and serological methods lack sensitivity, especially for pulmonary disease. In this study the capabilities of a new rapid test, the hydatid antigen dot immunobinding assay (HADIA), which was developed for the diagnosis of pulmonary hydatidosis, were studied and compared with another immunodiagnostic method, indirect hemagglutination (IHA). The study subjects included 18 patients, 9 women, 9 men; range 7 to 63 years; mean 30 years, with surgically proven pulmonary hydatidosis, a control group comprised of 14 patients; viral respiratory infections (1), cirrhosis (2), connective tissue disease (2), taeniasis (3), and 6 healthy donors. We found that the HA-DIA test had a sensitivity of 67% and specificity of 100%, and that the IHA test had a sensitivity of 50% and specificity of 100%. We conclude that HA-DIA is a simple, rapid, low cost assay that does not require instrumentation and has a higher sensitivity than IHA for the diagnosis of pulmonary hydatidosis. PMID- 15793355 TI - Protective efficacy of vaccination with Neospora caninum multiple recombinant antigens against experimental Neospora caninum infection. AB - Protective efficacy of vaccination with Neospora caninum multiple recombinant antigens against N. caninum infection was evaluated in vitro and in vivo. Two major immunodominant surface antigens (NcSAG1 and NcSRS2) and two dense granule proteins (NcDG1 and NcDG2) of N. caninum tachyzoites were expressed in E. coli, respectively. An in vitro neutralization assay using polyclonal antisera raised against each recombinant antigen showed inhibitory effects on the invasion of N. caninum tachyzoites into host cells. Separate groups of gerbils were immunized with the purified recombinant proteins singly or in combinations and animals were then challenged with N. caninum. Following these experimental challenges, the protective efficacy of each vaccination was determined by assessing animal survival rate. All experimental groups showed protective effects of different degrees against experimental infection. The highest protection efficacy was observed for combined vaccination with NcSRS2 and NcDG1. Our results indicate that combined vaccination with the N. caninum recombinant antigens, NcSRS2 and NcDG1, induces the highest protective effect against N. caninum infection in vitro and in vivo. PMID- 15793356 TI - Genetic characteristics of the Korean isolate KI-1 of Toxoplasma gondii. AB - Toxoplasma gondii tachyzoites were isolated from an ocular patient in the Republic of Korea and maintained in the laboratory (designated KI-1). In the present study, its genotype was determined by analyzing dense granule antigen 6 (GRA6) gene and surface antigen 2 (SAG2) gene as typing markers. Digestion of the amplification products of GRA6 and of the 5o and 3o ends of SAG2, respectively, with Mse I, Sau3A I, and Hha I, revealed that KI-1 is included in the genotype I, which includes the worldwide virulent RH strain. In addition, when the whole sequences of the coding regions of SAG1, rhoptry antigen 1 (ROP1), and GRA8 genes of KI-1 were compared with those of RH, minor nucleotide polymorphisms and amino acid substitutions were identified. These results show that KI-1 is a new geographical strain of T. gondii that can be included in the genotype I. PMID- 15793357 TI - Degranulation of human eosinophils induced by Paragonimus westermani-secreted protease. AB - Eosinophil degranulation is considered to be a key effector function for the killing of helminthic worms and tissue inflammation at worm-infected lesion sites. However, relatively little data are available with regard to eosinophil response after stimulation with worm-secreted products which contain a large quantity of cysteine proteases. In this study, we attempted to determine whether the degranulation of human eosinophils could be induced by the direct stimulation of the excretory-secretory products (ESP) of Paragonimus westermani, which causes pulmonary paragonimiasis in human beings. Incubation of eosinophils for 3 hr with Paragonimus-secreted products resulted in marked degranulation, as evidenced by the release of eosinophil-derived neurotoxin (EDN) in the culture supernatants. Moreover, superoxide anion was produced by eosinophils after stimulation of the ESP. The ESP-induced EDN release was found to be significantly inhibited when the ESP was pretreated with protease inhibitor cocktail or the cysteine protease inhibitor, E-64. These findings suggest that human eosinophils become degranulated in response to P. westermani-secreted proteases, which may contribute to in vivo tissue inflammation around the worms. PMID- 15793358 TI - Electroconvulsive therapy (ECT) in a patient with a dual-chamber sensing, VDDR pacemaker. AB - BACKGROUND: Because previous case reports involved old pacemaker models, it is unclear whether electroconvulsive therapy (ECT) may interfere with modern pacemaker function. Intracardiac voltages during ECT have not been measured previously. METHOD: We present a 77-year-old man with a dual-chamber sensing VDDR pacemaker who was treated for a major depression with ECT. His pacemaker function was closely monitored throughout the ECT sessions. RESULTS: An intracardiac electrogram showed low noise signals around 2 mV during ECT, which caused oversensing in the atrium. The intracardiac electrogram and pacemaker function normalized immediately after ECT. No arrhythmias or pacemaker malfunction was detected during and after several ECT sessions. CONCLUSION: The presence of a pacemaker should not be a contraindication to receiving ECT. There was no need to convert the pacemaker to a fixed rate pacing or deactivate the pacemaker before ECT. However, careful monitoring for potential pacemaker interference by medical staff experienced in pacemaker management is advised during the first ECT sessions for patients with pacemakers. PMID- 15793359 TI - Use of the serologic testing algorithm for recent HIV seroconversion (STARHS) to identify recently acquired HIV infections in men with early syphilis in Los Angeles County. AB - BACKGROUND: Syphilis outbreaks among men who have sex with men (MSM) in the United States, many of whom are HIV infected, have prompted increased concern for HIV transmission. METHODS: To identify whether men are acquiring HIV concomitantly or within the critical period of syphilis infection, banked Treponema pallidum particle agglutination-positive serum specimens from men with early syphilis infection were screened for HIV-1 antibody. Samples that were positive for HIV antibody were then tested with a less sensitive (LS) HIV-1 antibody enzyme immunoassay (serologic testing algorithm for recent HIV seroconversion [STARHS]) to identify HIV infections that occurred on average within the previous 6 months. RESULTS: Of the 212 specimens banked from men with early syphilis, 74 (35%) were HIV-positive. Of these, 15 tested non-reactive by the LS assay. Twelve of these 15 were considered to be recent infections by the LS assay and testing history. Eleven (92%) of the recent infections were among MSM. One man had primary syphilis, 6 (50%) had secondary syphilis, and 5 (42%) had early latent syphilis. Eight men (67%) reported sex with anonymous partners, and 3 (25%) reported consistent condom use. The estimated HIV incidence was 17% per year (95% confidence interval [CI]: 12%-22%) among all men with early syphilis, and it was 26% per year (95% CI: 91%-33%) among MSM. CONCLUSIONS: Syphilis epidemics in MSM may be contributing to HIV incidence in this population. The STARHS can be applied as a surveillance tool to assess HIV incidence in various at-risk populations, but further studies are necessary for validation. PMID- 15793360 TI - Functional analysis of naturally occurring mutations in the open reading frame of CCR5 in HIV-infected Chinese patients and healthy controls. AB - We studied polymorphism of the HIV coreceptor CC chemokine receptor (CCR) 5 in 1099 Chinese adults residing in Hong Kong, including 785 HIV-negative healthy donors and 314 HIV-positive patients. Ten mutants in the CCR5 open reading frame were identified, 7 of which were nonsynonymous. The frequencies of these alleles did not show a significant difference between HIV patients and healthy controls. G106R, Delta32, R223Q, 299(FS), and S336I were cloned from prevalent mutant genes, and their effects on HIV infection were analyzed by a series of in vitro experiments to determine their transcription levels, expression levels, conformational changes, and HIV coreceptor function. R223Q is the most prevalent CCR5 mutant in ethnic Chinese, with a frequency of 0.046, which does not affect HIV infection in vitro, however. The S336I mutant also does not affect its transcription, expression, or HIV coreceptor function. Similar to 299(FS), the mutant G106R located in the third transmembrane domain results in diminished HIV coreceptor function in vitro through conformation changes in ECL2. PMID- 15793361 TI - Active simian immunodeficiency virus (strain smmPGm) infection in macaque central nervous system correlates with neurologic disease. AB - Simian immunodeficiency virus strain smmPGm can induce neuropathology in macaques and is a model for the development of human HIV-related brain injury. For quantitative studies of proviral presence and expression in the central nervous system (CNS), we inoculated 8 macaques intravenously with the virus. Three animals were necropsied 2 to 4 weeks after development of infection, and we obtained lymphoid tissue biopsies from 5 animals before 5 weeks after infection. Peak plasma viral loads averaged 10 viral RNA Eq/mL at week 2, whereas cerebrospinal fluid viral loads peaked at 10 viral RNA Eq/mL. The proviral DNA loads and viral gag mRNA expression in tissues were quantified by real-time polymerase chain reaction. Two animals developed neurologic disease characterized by meningoencephalitis and meningitis. Proviral DNA levels in CNS tissues of these animals at necropsy revealed 10 and 10 copies/microg of DNA, respectively, whereas viral RNA expression in the CNS reached 100 to 1000 times higher levels than those seen in early necropsies. In sharp contrast, in 2 animals necropsied at later times without CNS disease, virus mRNA expression was not detected in any CNS tissue. Our results are consistent with the hypothesis that active virus expression in the CNS is strongly correlated with neurologic disease and that the event occurs at variable periods after infection. PMID- 15793362 TI - The unexpected movement of the HIV epidemic in the Southeastern United States: transmission among college students. AB - BACKGROUND: Approximately 16 million people are enrolled in institutions of higher learning in the United States. However, college students have not been perceived as at high risk for HIV infection. In early 2003, acute HIV infection was diagnosed in 2 men attending college in North Carolina. We describe an epidemiologic investigation of newly diagnosed HIV infection in men attending college in North Carolina. METHODS: We reviewed state surveillance records examining new HIV diagnoses in men 18-30 years old between January 1, 2000 and December 31, 2003, living in 69 North Carolina counties. Risk behavior and demographic information for HIV-infected men enrolled in college were compared with HIV-infected male nonenrollees. RESULTS: Of the 735 records available for review, 84 (11%) were college men. Eighty-seven percent of college men were African American and 92% were men who have sex with men (MSM) or men who have sex with men and women (MSM/W). Compared with noncollege men, college men were more likely to be African American (odds ratio 3.70, 95% CI = 1.86-7.54), to report meeting sex partners at bars or dance clubs (odds ratio 3.01, 95% CI = 1.77-5.10) or on the Internet/chat lines (odds ratio 4.95, 95% CI = 2.53-9.64), or to report use of "ecstasy" or club drugs (odds ratio 4.51, 95% CI = 1.15-15.40). Newly diagnosed HIV infection was found in men in 37 colleges located in North Carolina or surrounding states and a sexual partner network investigation linked 21 colleges, 61 students, and 8 partners of students. CONCLUSION: We describe an epidemic of HIV infection occurring in North Carolina college students, primarily involving African American MSM and MSM/W. College students represent an at-risk, accessible population, which deserves further HIV prevention interventions. PMID- 15793363 TI - Cytomegalovirus viremia, mortality, and end-organ disease among patients with AIDS receiving potent antiretroviral therapies. AB - OBJECTIVE: To determine the association of cytomegalovirus (CMV) viremia with CMV disease and death in patients with AIDS. DESIGN AND SETTING: Prospective, observational cohort study conducted at a university hospital. METHODS: A cohort of 190 subjects with AIDS who were CMV seropositive and had no history or evidence of CMV disease were longitudinally evaluated for signs and symptoms of CMV disease and CMV viremia with plasma CMV DNA polymerase chain reaction (PCR) and whole blood CMV hybrid capture. RESULTS: A total of 187 subjects had at least 1 study visit following entry. At baseline, the median CD4 cell count and plasma HIV RNA level were 110/microL (range = 3-620/microL) and 47,973 copies/mL (<30- >750,000 copies/mL), respectively. Highly active antiretroviral therapy (HAART) use increased from 87.5% during the 1st study year to 98.5% by the end of the study. During a median follow-up of 334 days, 16% (30) of the subjects died and 2 (6%) developed CMV disease. No deaths were attributable to CMV disease; 4 subjects who died developed CMV prior to death. Baseline HIV viral load and final CD4 cell count were significantly and independently associated with mortality. Detectable plasma CMV DNA PCR was an independent predictor of death even after adjusting for HIV RNA level and CD4 cell count prior to death (P = 0.038). In contrast, whole blood CMV hybrid capture did not predict mortality. The CMV assays neither collectively nor individually were found to be associated with the few cases of CMV disease. CONCLUSIONS: In patients with AIDS and seropositive for CMV, detection of CMV viremia with plasma CMV DNA PCR was predictive of death and provided additional prognostic information on the risk of all cause-mortality beyond that obtained with CD4 cell count and HIV viral load testing alone. Detection of CMV viremia by plasma with CMV DNA PCR in patients with AIDS, particularly those with low CD4 cell counts, provides additional rationale for optimization of antiretroviral therapy and consideration for preemptive anti-CMV therapy. PMID- 15793365 TI - A randomized controlled trial of the value of phenotypic testing in addition to genotypic testing for HIV drug resistance: evaluation of resistance assays (ERA) trial investigators. AB - OBJECTIVES: To assess the clinical utility of phenotypic resistance testing in addition to genotypic resistance testing among HIV-1-infected patients experiencing virologic failure and with limited therapeutic options. DESIGN: Multicenter randomized trial. METHODS: Patients were eligible if a decision had been made to switch antiretroviral therapy, the most recent HIV-1 RNA plasma viral load (VL) exceeded 2000 copies/mL, and the clinician was unable to select a potent regimen of 3 or more drugs without access to a resistance test. Subjects were randomized to genotypic resistance testing alone (G arm) or to genotypic plus phenotypic testing (G + P arm). Patients had access to resistance testing at any time during follow-up (minimum of 1 year) according to the original allocation. The primary end point was change in plasma VL from baseline at 12 months. RESULTS: Three hundred eleven patients were recruited between February 2000 and July 2001. At baseline, mean VL and CD4 count were 4.23 log10 copies/mL and 275 cells/mm, respectively, and subjects had previous exposure to a mean of 7.7 antiretroviral drugs. There was no appreciable difference between the study arms in the drug regimens prescribed after randomization. Mean reduction in VL load at 12 months was similar in the 2 arms (G: 1.37 log10 reduction, G + P: 1.28 log10 reduction; P = 0.77), as was the proportion of subjects with VL <50 copies/mL (G: 35%, G + P: 27%). CONCLUSION: The study did not demonstrate added value of phenotypic resistance testing in conjunction with genotypic resistance testing in patients with limited therapeutic options. PMID- 15793364 TI - French national sentinel survey of antiretroviral drug resistance in patients with HIV-1 primary infection and in antiretroviral-naive chronically infected patients in 2001-2002. AB - OBJECTIVE: To survey the frequency of genotypic antiretroviral resistance and the spread of non-B subtypes in patients with primary HIV-1 infection (2001-2002) and in treatment-naive chronically HIV-1-infected patients (2001). METHODS: Plasma samples from 303 patients with acute HIV-1 infection (Primo study) and 363 treatment-naive patients with chronic HIV-1 infection (Odyssee study) were tested for genotypic resistance. Resistance mutations were identified from the International AIDS Society Resistance Testing-USA panel and resistant viruses were defined according to the French Agence Nationale de Recherches sur le SIDA (ANRS) resistance algorithm. RESULTS: In the Primo study, 14% of the patients had viruses with resistance mutations and 12% of patients had viruses with mutations conferring resistance to least 1 antiretroviral drug. Thirty patients had viruses with mutations to at least 1 antiretroviral drug in a single pharmacologic class. Six patients were infected by viruses resistant to 2 or 3 classes of drugs. In the Odyssee study, the prevalence of reverse transcript (RT) associated and major protease inhibitor-associated mutations was 6.1% (95% CI: 3.6-8.6). Six patients had viruses resistant to at least 1 antiretroviral drug and 3 patients had viruses resistant to 2 classes of antiretroviral drugs. Twenty-four percent of acutely infected patients harbored non-B subtype strains (19% in 1999-2000) and 33.2% of chronically infected patients (10% in 1998; P < 0.0001). CONCLUSION: In France, the frequency of HIV-1 resistance in untreated patients was not significantly higher in 2001-2002 than in previous surveys while the prevalence of non-B subtypes is increasing. PMID- 15793366 TI - Long-term neuropsychiatric disorders on efavirenz-based approaches: quality of life, psychologic issues, and adherence. AB - BACKGROUND: Efavirenz has been associated with neuropsychiatric disorders, although little is known about its long-term toxicity. OBJECTIVE: To assess neuropsychiatric disorders and their relation to efavirenz plasma levels as well as quality of life, psychologic status, and adherence in HIV-infected patients on long-term efavirenz-based antiretroviral therapy. METHODS: Cross-sectional study comparing 60 patients on an efavirenz-based approach (EFV group) and 60 patients on a protease inhibitor-containing regimen (PI group) for at least 1 year. Adverse events, efavirenz plasma levels, quality of life, psychologic status, and adherence were assessed. RESULTS: The mean time on treatment was 91.1 +/- 39.5 weeks in the EFV group and 119.9 +/- 67.4 weeks in the PI group. Mild dizziness, sadness, mood changes, irritability, lightheadedness, nervousness, impaired concentration, abnormal dreams, and somnolence were reported more frequently in the EFV group than in the PI group (P < 0.05). Forty-nine of 60 patients presented with therapeutic efavirenz plasma levels (range: 1.0-4.0 mg/L). Efavirenz plasma levels were similar in subjects with and without neuropsychiatric disorders. No significant differences were found between the EFV group and the PI group regarding quality of life and psychologic status. Sixty percent of patients in the EFV group and 55% in the PI group reported adherence >/=95%. CONCLUSIONS: Mild and clinically tolerable neuropsychiatric disorders may persist in patients after a mean of 2 years using an efavirenz-based approach. Quality of life and psychologic status remained good in both study groups. Interventions to enhance long-term adherence should be applied in clinical practice. PMID- 15793368 TI - Potential cost-effectiveness of maternal and infant antiretroviral interventions to prevent mother-to-child transmission during breast-feeding. AB - INTRODUCTION: One-third of maternal-to-child HIV transmission occurs during breast-feeding (BF). Several trials are currently evaluating the efficacy of postpartum antiretrovirals to reduce BF transmission. METHODS: This study used Markov modeling to define the circumstances under which the following interventions would be cost-effective: BF for 6 months with daily infant nevirapine (NVP) prophylaxis; maternal combination antiretroviral therapy (ART) during pregnancy and for 6 months of BF; and maternal combination ART only for women who meet CD4 criteria. Each was compared to: BF for 12 months; BF for 6 months; and formula feeding for 12 months. Strategies were evaluated for a hypothetical cohort of 40,000 pregnant women in sub-Saharan Africa, in the context of available voluntary counseling and testing in antenatal care. Model estimates were derived from the literature and local sources. Sensitivity analyses were performed on uncertain estimates. The perspective used was that of a government health district. RESULTS: Using base case estimates, BF for 6 months was the economically preferred strategy: it cost 806,995 dollars and generated 446,208 quality-adjusted life-years (QALYs). Providing daily infant NVP cost an additional 93,638 dollars and generated 1183 additional QALYs, but its incremental cost-effectiveness ratio (ICER) of 79 dollars/QALY exceeded the standard willingness to pay (64 dollars/QALY) for most resource-poor settings. Maternal combination ART was potentially very effective but too costly for most resource-poor settings (ICER: 87 dollars/QALY). In order for daily infant NVP during BF to be preferred, it must have >/=44% relative efficacy or cost /=1 CD4 cell count or viral load done >/=90 days after starting therapy were evaluated. RESULTS: Three hundred twenty-one patients (49% women, 66% ARV naive, median age = 38 years, median CD4 cell count = 79 cells/mm, and median viral load = 249,489 copies/mL) attended the HIV clinic. Two hundred sixty-three (82%) patients returned at least once after the initial visit, of whom 54 (21%) had an interruption in therapy for >1 year. One hundred thirty-five patients were in care in 2002, 69 were known to have died (9 of whom died in 2002), and 68 were lost to follow-up. The probability of remaining alive and in care at 1 year was 0.56 (95% confidence interval [CI]: 0.50-0.61), 0.46 (95% CI: 0.41-0.51) at 2 years, 0.40 (95% CI: 0.34-0.45) at 3 years, and 0.35 (95% CI: 0.29-0.41) at 4 years. In an on treatment analysis, the median CD4 cell count increase during year 1 was +55 cells/mm, +112 cells/mm during year 2, +142 cells/mm during year 3, and +131 cells/mm during year 4. The median log viral load change from baseline during year 1 was -1.4 copies/mL, -1.32 copies/mL during year 2, -1.9 copies/mL during year 3, and -1.51 copies/mL during year 4. CONCLUSIONS: This fee-for-service HIV clinic providing ARV treatment has successfully operated and managed patients for more than 4 years. Those who survived and remained on therapy derived long-term virologic and immunologic responses to ARV drugs in a manner similar to that observed in industrialized countries. Strategies to reduce the financial burden and other barriers to uninterrupted care as well as incentives to increase such practice models should be further explored in the African context. PMID- 15793371 TI - Correlation of a brief perceived stress measure with nonadherence to antiretroviral therapy over time. AB - Adherence to antiretroviral medications has proven to be a challenge for individuals diagnosed with HIV infection. Nonadherence can lead to treatment failure, HIV resistance, and poor health outcomes. Many published studies have described factors associated with poor adherence, yet few have presented validated scales that could practically be applied in treatment settings to identify individuals at higher risk of nonadherence. This article explores the relationship between a revised version of the Perceived Stress Scale and nonadherence to antiretroviral therapy. The scale consists of the following items: How often in the past month have you felt that you were unable to control the important things in your life; confident in your ability to handle your personal problems; that things were going your way; and difficulties were piling up so high you could not handle them? Response options were "never or rarely," "sometimes," "often," and "mostly or always." In multivariate analysis, clients who scored in the highest quartile of perceived stress were more than twice as likely to be nonadherent at baseline and follow-up 1 and more than 5 times as likely to be nonadherent at follow-up 2 than clients in the lowest quartile of perceived stress scores. PMID- 15793370 TI - The effect of RANTES chemokine genetic variants on early HIV-1 plasma RNA among African American injection drug users. AB - HIV-1 plasma RNA is a prognostic indicator of HIV-1, and increased levels of HIV 1 plasma RNA are associated with rapid progression to AIDS. Because chemokines and chemokine receptors are involved in the binding and entry of HIV-1, possible effects of host genetics on viral RNA levels should be visible in early infection. HIV-1 plasma RNA was measured within 2 years of seroconversion in 198 seroincident injection drug users followed in the AIDS Link to Intravenous Experience cohort. Genetic variants were identified in the chemokine receptors (CCR2, CCR5, and CCR5 promoter) and the chemokine RANTES using TaqMan and restriction fragment length polymorphism assays. Linear regression of RANTES haplotypes on early HIV-1 plasma RNA identified individuals homozygous for the RANTES R1 haplotype as having a lower viral load by almost one-half log10 unit compared with those bearing non-RANTES R1 haplotypes (-0.43, 95% confidence interval: -0.74, -0.12). Genetic variants in RANTES may downregulate RANTES gene expression and increase early HIV-1 plasma RNA. Because RANTES is a critical chemokine and competitively inhibits HIV-1 by binding to its receptor CCR5, treatment to enhance RANTES expression may assist in delaying the progression of AIDS by decreasing the initial viral load. PMID- 15793372 TI - Mortality during the first 24 months after delivery in relation to CD4 T lymphocyte levels and viral load in a cohort of breast-feeding HIV-1-infected women in Dar es Salaam, Tanzania. AB - The objective of this study was to analyze the mortality during the first 24 months after delivery in relation to CD4 T-lymphocyte levels and viral load at enrollment (36 weeks of gestation) in a cohort of HIV-1-seropositive breast feeding women at the Dar es Salaam site of the multicenter Petra trial (a mother to-child HIV-1 transmission intervention trial using antiretroviral therapy). Antiretroviral treatment was not available in this setting apart from the short treatment given within the trial around delivery to prevent mother-to-child transmission of HIV. T-lymphocyte subsets were determined by flow cytometry. Plasma HIV-1 RNA was quantified by the Amplicor HIV-1 RNA Monitor v 1.5 assay. Mortality after delivery was analyzed using the life-table technique and Cox regression. The analysis included 266 mothers. The CD4 cell counts at enrollment were <200 cells/mm in 14.5% of the mothers. The viral load at enrollment was >100,000 RNA copies/mL in 33.6% of the mothers. The mortality 24 months after delivery was 6.7% (95% CI = 3.1-10.1%). The mortality 24 months after delivery was 29.9% (95% CI = 13.1-46.9%) for mothers with <200 CD4 cells/mm at enrollment, 3.3% (95% CI = 0-6.6%) for mothers with 200-499 CD4 cells/mm, 2.9% (95% CI = 0 7.1%) for mothers with >500 CD4 cells/mm (P = 0.0000), 15.0% (95% CI = 6.6-23.4%) for mothers with viral load >100,000 copies/mL at enrollment, and 2.8% (95% CI = 0-5.6%) for mothers with viral load <100,000 copies/mL (P = 0.0000). In the multivariate analysis CD4 cell counts and viral load were both independent risk factors for mortality (P < 0.001 and P = 0.004, respectively). In conclusion, the mortality was high among women with severe immunosuppression or high viral load at enrollment, but not in the rest of the women. CD4 lymphocyte count in late pregnancy was a better predictor of death within 2 years than was viral load. The results support the World Health Organization recommendation to initiate antiretroviral treatment in resource-limited settings in HIV-1-infected adults with CD4 cell counts <200/mm and show that this is appropriate also among perinatal women. PMID- 15793373 TI - Unrecognized HIV infection, risk behaviors, and perceptions of risk among young men who have sex with men: opportunities for advancing HIV prevention in the third decade of HIV/AIDS. AB - This study evaluated the magnitude and distribution of unrecognized HIV infection among young men who have sex with men (MSM) and of those with unrecognized infection, the prevalence and correlates of unprotected anal intercourse (UAI), perceived low risk for infection, and delayed HIV testing. MSM aged 15-29 years were approached, interviewed, counseled, and tested for HIV at 263 randomly sampled venues in 6 US cities from 1994-2000. Of 5649 MSM participants, 573 (10%) tested positive for HIV. Of these, 91% of black, 69% of Hispanic, and 60% of white MSM (77% overall) were unaware of their infection. The 439 MSM with unrecognized infection reported a total of 2253 male sex partners in the previous 6 months; 51% had UAI; 59% perceived that they were at low risk for being infected; and 55% had not tested in the previous year. The HIV epidemic among MSM in the United States continues unabated, in part, because many young HIV-infected MSM are unaware of their infection and unknowingly expose their partners to HIV. To advance HIV prevention in the third decade of HIV/AIDS, prevention programs must reduce unrecognized infection among young MSM by increasing the demand for and availability of HIV testing services. PMID- 15793374 TI - Dried blood spots improve access to HIV diagnosis and care for infants in low resource settings. AB - Effective health care delivery to the majority of perinatally exposed infants worldwide, including those enrolled in prevention of mother-to-child transmission programs, is hampered by lack of access to an HIV diagnosis in infancy. Dried blood spot collection from young infants for centralized HIV polymerase chain reaction (PCR) testing is attainable in low-resource settings, provided PCR methodology suitable for routine laboratory service is available. The accuracy of the Roche Amplicor HIV-1 DNA test version 1.5 (Branchburg, NJ) performed on dried blood spots collected prospectively on ordinary Whatman filter paper from a cohort of 300 6-week-old infants born to HIV-infected women in Johannesburg, South Africa, was assessed. Anonymous analysis of the blood spots using a unique DNA extraction procedure was performed in a routine diagnostic laboratory and the results compared with HIV DNA and RNA PCR liquid blood tests at age 6 weeks, and the HIV status of the infant. Dried blood spots were available for 288 infants (96%) of whom 25 (8.7%) were HIV infected. The Roche Amplicor assay yielded a sensitivity of 100% and a specificity of 99.6%. HIV DNA PCR tests on dried blood spots have the potential to improve health care delivery to HIV-affected children in low-resource settings right now. PMID- 15793375 TI - Behavior assessment of blood donors facing the risk of HIV infection, Abidjan, Cote D'Ivoire, 2001-2002. AB - Despite precautions taken to guarantee blood safety, in the National Blood Transfusion Center (CNTS) of Abidjan, about 30 regular blood donors are detected with HIV seroconversion each year, two-thirds of them men. A survey through face to-face interviews was carried out at the CNTS of Abidjan from September 2001 to March 2002 among HIV-positive and HIV-negative regular blood donors, informed about their serologic status. HIV-negative regular blood donors informed about their serologic status since a median time of 67 months (n = 50) disclosed more risky behaviors such as multiple sexual partners (68%) than HIV-positive blood donors informed about their status (n = 112) since a median time of 35 months (41%) (P < 0.001). Condoms were systematically used by 17% of HIV-negative blood donors and 55% of HIV-positive blood donors (P < 0.001). Enhanced counseling and awareness could reduce in the future the number of cases of seroconversion among regular blood donors and improve their subsequent behavior. Blood donors who have unprotected sex with partners of unknown HIV serologic status and especially with casual partners are strongly exposed to HIV transmission and should be discouraged to continue giving blood, after adequate counseling. PMID- 15793376 TI - Wasting during pregnancy increases the risk of mother-to-child HIV-1 transmission. AB - OBJECTIVES: To examine whether wasting during pregnancy, as measured by weight loss and low weight gain, is associated with increased mother-to-child transmission (MTCT) of HIV-1. METHODS: This was a cohort study in Dar es Salaam, Tanzania, among 957 HIV-1-infected pregnant women. Weight was measured at the first prenatal visit and every month thereafter until delivery. Weight loss was defined as a weekly rate of weight gain 0 and /=167 g/wk, weight loss during pregnancy was related to higher risk of intrauterine MTCT (adjusted relative risk [RR] = 2.32, 95% CI = 1.23-4.36, P = 0.009), HIV positive at birth or fetal death (RR = 2.13, 95% CI = 1.40-3.24, P = 0.0004), and HIV positive at birth or early neonatal death (RR = 1.96, 95% CI = 1.26-3.07, P = 0.003). The rate of weight gain during the 3rd trimester was inversely related to the risk of intrapartum/early breast-feeding transmission (adjusted P value, test for trend = 0.05). CONCLUSIONS: Weight loss during pregnancy increases the risk of early MTCT. Identifying causes of wasting during pregnancy may provide clues for new strategies to prevent MTCT. PMID- 15793377 TI - Efficacy of tenofovir as intensification of zidovudine/lamivudine/abacavir fixed dose combination in the treatment of HIV-positive patients. PMID- 15793378 TI - Cancer prevention and early diagnosis in HIV-positive individuals. PMID- 15793379 TI - The crisis in endodontic education: current perspectives and strategies for change. AB - The current status of dental education in the United States and Canada with respect to faculty recruitment and retention is reviewed; poor salaries and lack of interest are major reasons for dwindling numbers. The pros and cons of an academic career and what the American Association of Endodontists' (AAE) and other constituencies have done to help alleviate the problem are presented. The results of an on-line survey to full-time endodontic educators on their perspectives on education are presented; personal fulfillment, sharing knowledge, giving back to the profession, and passion for teaching were reasons for teaching whereas income disparity, lack of mentoring, and high demands of the job were reasons against an academic career. The final phase of the paper proposes various strategies on how the AAE and its membership, dental schools, and the American Board of Endodontics (ABE) can further work together to alleviate the critical problems facing endodontic education. PMID- 15793380 TI - A 5 yr clinical investigation of second mesiobuccal canals in endodontically treated and retreated maxillary molars. AB - An examination of 5616 endodontically treated and retreated maxillary first and second molars was made in an attempt to determine the percentage of MB2 canals that could be located routinely, and evaluate if there were any significant differences between initial treatments and retreatments. The teeth examined were 3578 first molars and 2038 second molars treated consecutively over a 5-yr period by six endodontists. Overall the MB2 canal was found in 2133 (60%) first molars, and 712 (35%) second molars. The incidence of a MB2 canal in first molar retreatments was 66% compared to a 58% incidence in initial treatments. Whereas in second molars the retreatment incidence was 40% compared to 34% in initial treatments. The significant difference in the incidence of a MB2 canal between initial treatments and retreatments suggests that failure to find and treat existing MB2 canals will decrease the long-term prognosis. PMID- 15793381 TI - A comparison of articaine and lidocaine for inferior alveolar nerve blocks. AB - The purpose of this prospective, randomized, double-blind study was to compare the degree of pulpal anesthesia obtained with 4% articaine with 1:100,000 epinephrine and 2% lidocaine with 1:100,000 epinephrine in inferior alveolar nerve blocks. Using a crossover design, inferior alveolar nerve blocks were randomly administered, in a double-blind manner, using 4% articaine with 1:100,000 epinephrine and 2% lidocaine with 1:100,000 epinephrine, at two separate appointments, to 57 subjects. A pulp tester was used to test for anesthesia, in 4-min cycles for 60 min, of the molars, premolars, central, and lateral incisors. Anesthesia was considered successful when two consecutive 80 readings were obtained within 15 min and the 80 reading was continuously sustained for 60 min. Using the articaine solution, successful pulpal anesthesia ranged from 4 to 54% from the central incisor to the second molar. Using the lidocaine solution, successful pulpal anesthesia ranged from 2 to 48%. There was no significant difference (p > 0.05) between the articaine and lidocaine solutions. We concluded that 4% articaine with 1:100,000 epinephrine was similar to 2% lidocaine with 1:100,000 epinephrine in inferior alveolar nerve blocks. PMID- 15793382 TI - Determining the optimal obturation length: a meta-analysis of literature. AB - The purpose was to aid in determining termination of instrumentation and obturation. A meta-analysis was conducted as to success/failure of different obturation lengths. Inclusion criteria were (a) minimum follow-up of 2 yr, (b) data on obturation length, (c) definition of success/failure, (d) available data on success/failure, (e) radiographic evaluation. Correlations were made as to success/failure as related to length of obturation from the apex. When comparing group A (obturated 0-1 mm from apex) versus group C (obturated past apex) using the DerSimonian and Laird estimates, group A showed a marginally better (p < 0.10) success rate than group C by 28.8%. Group A had better success than group B (obturated >1 mm short); the difference was insignificant. The results were similar after controlling for study quality using a single random effects regression model. In conclusion, the meta-analysis indicated that a better success rate is achieved when treatment includes obturation short of the apex. PMID- 15793383 TI - Preliminary evaluation of BMP-2 expression and histological characteristics during apexification with calcium hydroxide and mineral trioxide aggregate. AB - Histological characteristics and bone morphogenetic protein-2 (BMP-2) expression were evaluated in apexification procedures using calcium hydroxide [Ca (OH)2] and mineral trioxide aggregate (MTA). In three of four quadrants of the mouth, monkey (Macaca fascicularis) teeth were infected by pulpal exposure to saliva. After 90 days, each group was treated with Ca (OH)2 or MTA. Infected, untreated teeth were used as negative controls and normally developing teeth were used as positive controls. Areas within the apical region of each sample were scored based upon histological findings and BMP-2 expression. Overall, BMP-2 expression in all areas was similar in the Ca (OH)2, MTA, and infection/no treatment groups, which tended to be more immunoreactive for BMP-2 than the untreated group. Use of MTA in combination with Ca (OH)2 may initiate regeneration of the periodontium more quickly than either material used alone, but further studies involving MTA use in apical closure are indicated. PMID- 15793384 TI - Coronal leakage of sealer-only backfill: an in vitro evaluation. AB - We tested the hypothesis that AH Plus and Roeko Seal Automix (RSA) sealers alone are no less effective in preventing coronal microleakage than gutta-percha compacted with sealer. Freshly prepared sheep incisor root canals were obturated with warm gutta-percha alone, AHPlus or RSA alone, or warm gutta-percha with AH Plus or RSA (n = 20 each group). Coronal leakage was assessed under vacuum conditions with Indian ink and tooth clearing. Sealer-only backfills with AH Plus and RSA yielded significantly more dye-free canals than backfills of gutta-percha alone or with sealer (p < 0.001). Warm gutta-percha with or without sealer kept no more canals sealer free than the positive control. Mean dye penetration was 0.92% of canal length for AH Plus and RSA backfills, 27.42% for gutta-percha only backfills, 26.47% for gutta-percha with RSA and 13.92% for gutta-percha with AH Plus. Sealer only backfills allowed significantly less leakage than those including warm gutta-percha (p < 0.001). Sealer-only backfills may be a viable alternative to traditional gutta-percha and sealer compaction methods. PMID- 15793385 TI - A comparison of the effects of two canal preparation techniques on root fracture susceptibility and fracture pattern. AB - The purpose of the study was to determine whether rotary nickel-titanium (NiTi) canal preparation strengthens roots, and whether the fracture pattern can be predicted by finite element analysis (FEA) models. From a fracture mechanics viewpoint, structural defects, cracks or canal irregularities are likely to play a major role in fracture susceptibility of the roots, because stresses can be exponentially amplified at these sites. By incorporating defects into a smooth round canal using rotary NiTi, theoretically the roots could be strengthened. 25 teeth were prepared using hand files and another 25 using rotary NiTi. After obturation, all teeth were subject to loading until fracture; load and patterns were recorded. Four FEA models were created from fractured roots. No significant difference of fracture load between the two techniques was found. Mesio-distal fracture occurred more often in the rotary NiTi group. Stress patterns in three of the four FEA models correlated well with the observed fracture patterns. PMID- 15793386 TI - Effect of root canal size and external root surface morphology on fracture susceptibility and pattern: a finite element analysis. AB - The aim of the study was to determine the extent to which canal size, radius of curvature and proximal root concavity influence fracture susceptibility and pattern. A standardized cross-section of the mid-root region of a mandibular incisor was created by averaging the dimensions of 10 extracted teeth, and then the basic finite element analysis (FEA) model was created. By varying canal diameter, shape, and proximal concavity, these factors could be examined for roles in fracture susceptibility and pattern. The factors all interact in influencing fracture susceptibility and pattern, with dentin thickness not the only determining factor. The removal of dentin does not always result in an increased fracture susceptibility. PMID- 15793387 TI - Shear bond strength of three resin based sealers to dentin with and without the smear layer. AB - Bond strength of root canal sealers to dentin is an important property for the integrity of the sealings of root-canals. The purpose of this study was to test shear bond strength of three endodontic sealers (Diaket, AH Plus and Endo-REZ). The coronal two thirds of ninety extracted human third molars were removed. The smear layer of the exposed dentin surfaces were removed using 17% EDTA followed by 5.25% NaOCl and the teeth were randomly divided into two groups (n = 45). Group 1 was kept as control and in group 2, uniform smear layer was created using waterproof polishing papers. Three-mm long sections of polyethylene tubing were filled with freshly mixed sealer and placed on the dentin surfaces for conducting a shear bond strength test. The data was calculated as MPa and was statistically analyzed using a two way ANOVA, Kruskal-Wallis and Mann-Whitney U tests. A significant difference was found among the bond strength of the sealers, smear layer, and control groups. AH Plus sealer showed the highest bond strength in smear layer removed surfaces (p < 0.05). Pretreatment with EDTA/NaOCl affected bond strength of AH Plus. AH Plus had the highest bond to dentin with or without smear layer. PMID- 15793388 TI - Root surface temperature rises in vitro during root canal obturation using hybrid and microseal techniques. AB - The aim of this in vitro study was to measure the temperature rise on the outer surface of roots during filling with hybrid technique and Microseal. Twenty extracted human maxillary and mandibular premolars with a single canal were randomly divided into two groups of 10 teeth each. In the first group, the teeth were filled with hybrid technique (thermomechanical compaction with Engine Plugger used following lateral condensation of the apical part of the canal), the second group was filled using Microseal. After root canal obturation the filling material was removed and the obturation procedure was repeated. A total of 20 obturations in each group were performed. Temperature changes were measured using a thermal imaging camera. The temperature of certain regions of the mesial surface was analyzed and the highest temperature values were recorded. Statistical analysis was performed using the Mann-Whitney U test. The mean increase of temperature during the hybrid technique with Engine Plugger was 23.8 degrees C, while during Microseal it was significantly lower (p = 0.000001) at 5.5 degrees C. The temperature rise generated by Microseal was below the critical level and should not damage supporting structures, however, the hybrid technique generated a relatively high temperature rise that may cause periodontal tissue damage. PMID- 15793389 TI - Analysis of heat generation using ultrasonic vibration for post removal. AB - This study measured the temperature of the root surface and post during the application of ultrasonic vibration to cemented posts to simulate post removal procedure. Root canal therapy was performed on ten extracted maxillary incisors. A stainless steel Parapost was cemented into each prepared post space. Ultrasonic vibration was applied to the post and temperatures were recorded at the coronal post and the cervical root surface. Data were analyzed with ANOVA using the independent variables of (a) time of ultrasonic application (15, 30, 45 and 60 s) and 2) location (post and root surface). Greater temperature increase was observed at the post (52.6 degrees C, SD 11.1; 82.6 degrees C, SD 20.1; 111.0 degrees C, SD 29.1; 125.3 degrees C, SD 33.2) compared to the root surface (9.5 degrees C, SD 4.6; 17.5 degrees C, SD 4.8; 25.4 degrees C, SD 7.3; 32.2 degrees C, SD 8.1) for each time period, P < 0.001. Ultrasonic application to the post for longer than 15 s generates high temperature on the root surface. PMID- 15793390 TI - Comparison of two different direct digital radiography systems for the ability to detect artificially prepared periapical lesions. AB - The purpose of this study was to compare Schick CDR and Trophy RVGui direct digital radiography (DDR) systems for the ability to detect periapical lesions in human cadaver mandibles. Digital radiographs were exposed of teeth with normal periapical areas and of teeth with artificially prepared periapical lesions using both DDR systems. Three examiners independently viewed the images at two different time periods and estimated which bony state was present. The resulting data were subjected to statistical analysis using a two-way ANOVA. Interexaminer variability was statistically analyzed using Spearman's rho. There was no significant difference in the level of accuracy between the two different DDR systems at either observation period. There was a statistically significant high level of agreement between examiners (p < 0.01). In conclusion, there was no significant difference in the accuracy of detecting artificially prepared periapical lesions between Schick CDR and Trophy RVGui DDR systems. PMID- 15793391 TI - The cytotoxicity of RoekoSeal and AH plus compared during different setting periods. AB - The aim of this study was to evaluate the cytotoxicity of the root canal sealers RoekoSeal Automix (RSA) and AH Plus in vitro on human cervical carcinoma (HeLa) cells and mouse skin fibroblasts (L929). The sealers were covered with a 3 x 10(4) cell suspension 1 h, 24 h, 48 h, 7 days and 1 month after mixing. Four samples and respective controls without sealer were prepared. After 5 days of incubation, the number of cells was determined using an electronic counter; the number of viable cells was determined under light microscopy following the addition of nigrosin dye. The experiment was repeated twice for each sealer and each period. AH Plus was significantly more cytotoxic after 1 h, 24 h, and 48 h, compared to the 7 day and 1 month setting period on both cell lines. RoekoSeal had no cytotoxic effect on either cell line at any setting time. PMID- 15793392 TI - Root canal treatment in a mandibular second premolar with three root canals. AB - According to the endodontic literature, the possibility of three canals in mandibular second premolars is quite small. When three or more canals are present separation of the root canals has only been described at mid-root level. The present case describes root canal treatment of a root canal aberration at the level of the apical third (apical root canal separation starting at a length of 20 mm) where a combined use of the fiberoptic endoscope and the operating microscope was necessary to locate the canal orifices and to enter these orifices. The present report emphasizes the need of color contrast to enhance visualization. PMID- 15793394 TI - Efficacy of the KTP laser in the treatment of middle ear cholesteatoma. AB - OBJECTIVE: To evaluate whether ancillary use of the KTP (potassium titanyl phosphate) laser can diminish the rate of residual disease in intact canal wall cholesteatoma surgery. STUDY DESIGN: Prospective controlled parallel group study. SETTING: Two neighboring district general hospitals, one with a KTP laser and one without. PATIENTS: All patients undergoing staged intact canal wall surgery for cholesteatoma. INTERVENTION: Staged intact canal wall surgery for cholesteatoma. MAIN OUTCOME MEASURE: Presence or absence of residual disease at the second stage operation, performed at least 12 months after the first operation. RESULTS: Thirty-three patients underwent treatment without and 36 underwent treatment with the laser. Ten patients without laser treatment had residual disease, whereas one patient had residual disease after laser treatment (p = 0.003). After adjustment using logistic regression, treatment still has a significant effect on outcome (p = 0.013). The number needed to treat is four. DISCUSSION: Ancillary use of the KTP laser in cholesteatoma surgery is a treatment that significantly improves complete removal of disease. PMID- 15793395 TI - Otologic effects of topical mitomycin C: phase I-evaluation of ototoxicity. AB - HYPOTHESIS: To determine ototoxicity of topical mitomycin C when placed in the middle ear at varying concentrations. BACKGROUND: Despite meticulous surgical technique and diligent postoperative care, some patients develop excessive scar and granulation tissue in the middle ear or mastoid cavity. Poor wound healing may result in infection, tympanic membrane perforation, or conductive hearing loss, which may necessitate further surgery. Use of topical mitomycin C in the ear may be beneficial in reducing scar and granulation tissue formation. This phase of the study was developed to determine the safety of topical mitomycin C in the rat model relative to ototoxicity. METHODS: Twelve Sprague-Dawley rats were evaluated with auditory brainstem response testing before and after treatments. Topical mitomycin C was injected in the middle ear of the right ear of eight animals. Varying concentrations of 0.125 to 0.5 mg/ml were used. Saline was injected in the left ear of each animal to serve as a control. Four separate animals were evaluated with placement of topical mitomycin C on Gelfoam into the middle ear. In two animals, Gelfoam was placed in the middle ear for 1 minute and then removed. In two animals, Gelfoam was placed in the middle ear and left in place. Auditory brainstem response testing was performed at 4 weeks and at 8 weeks. RESULTS: Using a high concentration of mitomycin C (>0.25 mg/ml) resulted in ototoxicity, with an increase in the auditory brainstem response threshold at 4 weeks and at 8 weeks. At low concentrations (<0.20 mg/ml), no change in auditory brainstem response threshold was noted. Animals treated with Gelfoam soaked in mitomycin C showed no change in auditory brainstem response threshold. CONCLUSION: The results of this study indicate that topical mitomycin C on Gelfoam applied in the middle ear appears safe when low concentrations are used, even in the rat, which has a higher susceptibility to gentamycin toxicity than humans. Higher concentrations may lead to ototoxicity based on changes in Wave V on auditory brainstem response. This treatment may prove to be an important option for patients suffering from chronic granulation tissue or scar tissue in the external or middle ear. PMID- 15793396 TI - Otologic disease in turner syndrome. AB - OBJECTIVE: This study involved the assessment of the otologic and audiologic characteristics of a group of patients with Turner syndrome. STUDY DESIGN: Prospective study consisting of a questionnaire (77 of 123 responders) and an otologic and audiologic evaluation in patients with Turner syndrome (41 participants). SETTING: Tertiary academic medical setting. PATIENTS: Children, adolescents, and adults with Turner syndrome (median age, 24 yr). INTERVENTIONS: Otomicroscopy, audiometry, immittance measurements, and diagnostic imaging. RESULTS: Otologic disease is an important characteristic in Turner syndrome. Sixty-six percent of the patients studied via the questionnaire reported a history of chronic or recurrent middle ear disease. Analysis of audiometric data in 40 patients tested reveals an equal amount of normal ears (38.8%) and pure sensorineural ears (38.8%), each constituting approximately one-third of the patient population. Pure conductive losses represent only one-fifth (21.3%) of auditory abnormality encountered. CONCLUSIONS: Careful follow-up during early childhood of children with Turner syndrome is necessary to detect middle ear disease and prevent sequelae. However, long-term periodic review is mandatory even after resolution of middle ear disease to detect sensorineural hearing loss. PMID- 15793397 TI - Pathology and pathophysiology of idiopathic sudden sensorineural hearing loss. AB - BACKGROUND: The cause and pathogenesis of idiopathic sudden sensorineural hearing loss remain unknown. Proposed theories include vascular occlusion, membrane breaks, and viral cochleitis. AIMS: To describe the temporal bone histopathology in 17 ears (aged 45-94 yr) with idiopathic sudden sensorineural hearing loss in our temporal bone collection and to discuss the implications of the histopathologic findings with respect to the pathophysiology of idiopathic sudden sensorineural hearing loss. METHODS: Standard light microscopy using hematoxylin and eosin-stained sections was used to assess the otologic abnormalities. RESULTS: Hearing had recovered in two ears and no histologic correlates were found for the hearing loss in both ears. In the remaining 15 ears, the predominant abnormalities were as follows: 1) loss of hair cells and supporting cells of the organ of Corti (with or without atrophy of the tectorial membrane, stria vascularis, spiral limbus, and cochlear neurons) (13 ears); 2) loss of the tectorial membrane, supporting cells, and stria vascularis (1 ear); and 3) loss of cochlear neurons only (1 ear). Evidence of a possible vascular cause for the idiopathic sudden sensorineural hearing loss was observed in only one ear. No membrane breaks were observed in any ear. Only 1 of the 17 temporal bones was acquired acutely during idiopathic sudden sensorineural hearing loss, and this ear did not demonstrate any leukocytic invasion, hypervascularity, or hemorrhage within the labyrinth, as might be expected with a viral cochleitis. DISCUSSION: The temporal bone findings do not support the concept of membrane breaks, perilymphatic fistulae, or vascular occlusion as common causes for idiopathic sudden sensorineural hearing loss. The finding in our one case acquired acutely during idiopathic sudden sensorineural hearing loss as well as other clinical and experimental observations do not strongly support the theory of viral cochleitis. CONCLUSION: We put forth the hypothesis that idiopathic sudden sensorineural hearing loss may be the result of pathologic activation of cellular stress pathways involving nuclear factor-kappaB within the cochlea. PMID- 15793398 TI - Autoimmune-mediated sympathetic hearing loss: a case report. AB - BACKGROUND: Damage to one inner ear is occasionally followed by contralateral sensorineural hearing loss. This has been defined as sympathetic hearing loss. HYPOTHESIS: It is hypothesized that autoimmunity can play a role in the pathogenesis of sympathetic hearing loss. METHODS: A male patient who developed right-sided sympathetic hearing loss at 20 years of age, 11 years after deafness of the left ear caused by a temporal bone fracture, is described. The patient's serum was analyzed for the presence of autoantibodies against inner ear tissues by immunocytochemistry and Western blotting using rat inner ear tissues. The patient's serum was tested specifically for antibodies against heat shock protein 70 by immunodot blot. The presence of autoantibodies known to play a role in systemic autoimmune disease was also examined. RESULTS: Immunocytochemistry on rat temporal bone sections demonstrated autoantibodies in the patient's serum specifically targeted against cochlear outer hair cells. No reactivity of the patient's serum was observed with control tissues including kidney, brain, and liver. Western blotting using homogenized rat cochlear tissues showed that the patient's serum reacted with a 25- and 27-kDa protein. No reactivity was observed with heat shock protein 70 in the immunodot blot analysis. The patient's serum did not contain autoantibodies against antinuclear antibodies, double-stranded DNA, antineutrophil cytoplasmic antibodies, basement membrane, reticulin, intestinal mucosa, muscle, collagen, or mitochondria. CONCLUSION: Observations indicate that this patient suffered sympathetic hearing loss caused by organospecific autoimmunity directed to cochlear outer hair cells. PMID- 15793399 TI - Pulsatile tinnitus and spontaneous cerebrospinal fluid rhinorrhea: indicators of benign intracranial hypertension syndrome. PMID- 15793400 TI - The inhibitory effect of intravenous lidocaine infusion on tinnitus after translabyrinthine removal of vestibular schwannoma: a double-blind, placebo controlled, crossover study. AB - OBJECTIVE: Intravenous infusion of lidocaine has previously been demonstrated to have a transient inhibitory effect on tinnitus in 60% of individuals. The site of action has variously been proposed as the cochlea, the cochlea nerve, and the central auditory pathways. To determine whether a central site of action exists, this study investigated the effect of intravenous infusion of lidocaine in individuals with tinnitus who had previously undergone translabyrinthine excision of a vestibular schwannoma, which involves division of the cochlear nerve. STUDY DESIGN: Double-blind, placebo-controlled, crossover study. SETTING: University hospital. PATIENTS: Patients who had undergone translabyrinthine removal of a unilateral, sporadic, and histologically proven vestibular schwannoma in the last decade and who had reported postoperative tinnitus at follow-up were identified from a departmental database. Sixteen patients participated (12 men and 4 women). The mean age (+/- standard deviation) of the patients was 58 +/- 8.6 years, and the meantime since operation was 24.3 +/- 7.3 months. INTERVENTION: Solutions of 2% lidocaine hydrochloride and sodium chloride 0.9% were prepared in identical randomized vials. The volume required for 1.5 ml/kg body weight lidocaine was calculated, and this volume was given over 5 minutes for either vial. Blood pressure, pulse oximetry, and cardiac monitoring were set up and performed throughout the infusions. All investigators were blinded. OUTCOME MEASURES: Patient-completed visual analogue scale measures of tinnitus intensity, pitch, and distress, performed before infusion, 5 minutes after infusion onset, and 20 minutes after infusion onset. RESULTS: A significant difference (Wilcoxon signed rank test, p < 0.05) between placebo and lidocaine infusion conditions was demonstrated for change in visual analogue scale estimates (preinfusion versus 5 min postinfusion) of tinnitus loudness (p = 0.036), pitch (p = 0.026), and distress (p = 0.04). No significant difference between placebo and lidocaine infusion conditions was demonstrated for change in visual analogue scale estimates (preinfusion versus 20 min postinfusion) of tinnitus loudness (p = 0.066), pitch (p = 0.173), and distress (p = 0.058). The indication is of a short lasting inhibitory effect on tinnitus of lidocaine infusion compared with saline placebo in patients who have undergone translabyrinthine excision of a vestibular schwannoma. CONCLUSION: Intravenous infusion of lidocaine has a statistically significant inhibitory effect on tinnitus in patients who have previously undergone translabyrinthine removal of a vestibular schwannoma. The site of action of lidocaine in this instance must be in the central auditory pathway, as the cochlear and vestibular nerves are sectioned during surgery, and this finding has important implications for the task of identifying other agents that will have a similar tinnitus-inhibiting effect. PMID- 15793401 TI - Double posterior labyrinthotomy technique: results in three Med-El patients with common cavity. AB - HYPOTHESIS: This study reports on the use of the double posterior labyrinthotomy surgical technique and a custom-designed electrode to ensure better positioning of stimulating electrodes within the common cavity and thus demonstrate suitable outcomes in patients. BACKGROUND: Cochlear implantation has proven beneficial for numerous children with congenital malformations of the inner ear. Several studies show good auditory perception outcomes in children with common cavity. However, there have been risks involved with surgical techniques used in the actual implantation. These include possible aberrant facial nerve and the strong potential for a cerebrospinal fluid gusher. Improved surgical techniques and electrode design could allow for better electrode contact and avoid electrode placement in the internal auditory meatus. METHOD: The double posterior labyrinthotomy technique was carried out in three cases using a custom made MED EL COMBI 40+ electrode. RESULTS: Surgery was carried out with no complications and is no more technically demanding than other standard surgical approaches. The speech processor program remains stable over time, and auditory perception results are similar to those obtained from children with no cochlear abnormalities. CONCLUSION: These results demonstrate the success of the double posterior labyrinthotomy approach with modified cochlear implant, and this could be recommended as the procedure of choice in children presenting to an implant team with a common cavity. PMID- 15793403 TI - Quality-of-life benefit from cochlear implantation in the elderly. AB - OBJECTIVE: To compare the audiologic results of geriatric patients receiving cochlear implants with younger age groups and to evaluate the quality of life after cochlear implantation in the geriatric population by means of validated quality-of-life questionnaires. STUDY DESIGN: Cross-sectional study involving 89 postlingually deafened cochlear implant subjects. SETTING: Tertiary referral center. PATIENTS: A total of 89 postlingually deafened patients were included in the study, among which were 25 patients who were aged 70 years or older. INTERVENTIONS: All patients received a cochlear implant. Subjects were implanted with either the Laura, Nucleus 24, or Med-el Combi 40+ cochlear implant systems implementing the SPEAK, ACE, CIS, or CIS+ coding strategies. MEAN OUTCOME MEASURES: Speech recognition was determined by means of phonetically balanced monosyllabic word lists. The Hearing Handicap Inventory for Adults, the Glasgow Benefit Inventory, and the scale for the prediction of hearing disability in sensorineural hearing loss were used to quantify the quality of life. RESULTS: Mean audiologic performance for the three groups increased significantly after implantation (p < 0.001). Postoperative audiologic performance of the geriatric population led to useful hearing, but these scores were significantly lower than for the younger age groups (p = 0.002). However, the quality-of-life outcomes for the geriatric group were similar to those of the younger age groups (p = 0.411 for the Hearing Handicap Inventory for Adults; p = 0.886 for the Glasgow Benefit Inventory). CONCLUSION: The results of this study prove that cochlear implantation in the elderly provides improvements in quality of life and speech understanding, similar to those for younger adult cochlear implant recipients. PMID- 15793402 TI - Device nonuse among adult cochlear implant recipients. AB - OBJECTIVE: To examine the causes and prevalence of previous and current device nonuse among adults who have received cochlear implants. STUDY DESIGN: Retrospective case review. SETTING: Adult tertiary referral center for cochlear implantation. METHODS: Two hundred fourteen consecutively implanted adult patients. The length of implant use ranged from 1 month to 14 years. MAIN OUTCOME MEASURE: A period of 4 consecutive weeks of nonuse of cochlear implant, including both obligatory and elective nonuse. RESULTS: Twenty-nine adults (13.6% of implantees) were identified as having at some stage not used their device for a period of more than 4 consecutive weeks. The main reason was device failure (n = 11). Ten adults are current nonusers (4.7% of implantees). Reasons include surgical complication necessitating explantation (n = 3), comorbid illness (n = 3), elective nonuse (n = 2), audiologic complication (n = 1), and device failure (n = 1). CONCLUSION: The overall prevalence of device nonuse was noted to increase slowly with time. The role of psychologic factors in contributing to the decision of an individual to elect to opt out of device use remains unproven. PMID- 15793404 TI - Chronic Pseudomonas infections of cochlear implants. AB - OBJECTIVE: To discuss chronic, refractory Pseudomonas infections of cochlear implants and their management. DESIGN: Retrospective case series. SETTING: Two university-based cochlear implant programs. PATIENTS: Twenty-eight-year-old (Case 1) and 4-year-old (Case 2), different devices. INTERVENTIONS: Medical and surgical management. MAIN OUTCOME MEASURES: Clinical course. RESULTS: Both patients had delayed presentations, 4 months and 3 years postimplantation, respectively, with fluctuating scalp edema and pain resistant to multiple courses of oral antibiotics. Infections began as localized granulation and progressed to complete encasement of both devices with rubbery, poorly vascularized tissue. In each case, two different strains of multiresistant Pseudomonas aeruginosa were cultured. Infections progressed despite local debridement and targeted antipseudomonal antibiotic coverage, and sensitive organisms continued to appear in cultures of refractory granulation tissue. Both patients underwent partial explantation, with the electrode array left in the cochlea, then received 2 to 3 more months of further medical therapy and observation and then were reimplanted successfully with new devices. Both have shown excellent performance and no sign of recurrent infection. CONCLUSIONS: Infections of cochlear implants are uncommon, and cases of successful conservative management without device explantation have been reported. However, our experience and the implanted device literature suggest that chronic Pseudomonas infections may represent a distinct clinical entity, likely to fail protracted therapy and ultimately require device removal. Fortunately, successful reimplantation is possible. PMID- 15793405 TI - Epidural hematoma after cochlear implantation in a 2.5-year-old boy. AB - OBJECTIVE: Report a case of an epidural hematoma after cochlear implantation in a 2.5-year-old boy, the diagnostic and therapeutical emergency management, as well as the postinterventional course and rehabilitation of the child. STUDY DESIGN: Retrospective case review. PATIENT: Two and a half-year-old boy, suffering from early onset, profound sensorineural hearing loss had been diagnosed at an age of 1.5 years, which had been more severe on the right side initially, but had progressed to bilateral deafness. INTERVENTION AND COMPLICATION: Cochlear implantation on the left side, followed up by an extensive epidural hematoma, causing intracranial compression with a midline shift of 15 mm to the right side. This initial complication was followed up by arterial infarction not only of the middle but also the posterior as well as the anterior cerebral artery, most likely caused by incarceration through the tentorium. RESULTS: After immediate neurosurgical intervention and intensive care treatment including low-dose anticoagulation and high doses of corticosteroids, the child recuperated completely within 3 weeks without any remaining neurologic deficits. CONCLUSION: In cochlear implantation, especially if placement of the implant housing is associated with considerable bone work, epidural hematoma has to be considered as an extremely rare, but life-threatening, complication. PMID- 15793406 TI - Simultaneous cochlear implantation and translabyrinthine removal of vestibular schwannoma in an only hearing ear: report of two cases (neurofibromatosis type 2 and unilateral vestibular schwannoma). AB - OBJECTIVE: The goal is to confirm the effectiveness of cochlear implantation performed at the time of surgery for tumor removal, using a translabyrinthine approach. STUDY DESIGN: This is a retrospective case review of two cases of vestibular schwannoma (VS) in the only hearing ear in which a cochlear implant (CI) was placed simultaneous to removal of VS through a modified enlarged translabyrinthine approach. The aim of the study is to confirm the effectiveness in terms of hearing rehabilitation. SETTING: The study involved a tertiary referral center in a hospital setting. PATIENTS: The study includes a report of two cases: a case of a Neurofibromatosis Type 2 (NF2) patient who received a MEDEL COMBI 40+ CI after a translabyrinthine surgery for removal of a large (4.0 cm) cystic VS; and a patient with a 1.2 cm VS in the only hearing ear who was submitted to the same strategy and operation, using a Nucleus 24 Contour CI. INTERVENTIONS: The whole strategy used in the two mentioned cases is outlined, including decision making, surgical interventions, and hearing rehabilitation programs. MAIN OUTCOME MEASURES: Surgical and audiometric outcome of two patients who underwent VS resection through a translabyrinthine approach and simultaneous cochlear implantation are shown. RESULTS: Postoperative results demonstrate that a CI could be successful after a translabyrinthine surgery for VS, with hearing performances similar to the best postlingual implanted deaf adults of other origin. CONCLUSION: The present cases demonstrate that cochlear implantation can be successful after a translabyrinthine approach for VS, regardless of the tumor size, the kind of patient (NF2, unilateral VS), and the type of implant. The results also are suggestive that cochlear implantation is more successful if done concurrent with surgery for tumor removal and before hearing is completely lost. PMID- 15793408 TI - The effects of mass loading the ossicles with a floating mass transducer on middle ear transfer function. AB - HYPOTHESIS: The aim of this study was to measure the mass loading effect of an active middle-ear implant (the Vibrant Soundbridge) in cadaver temporal bones. BACKGROUND: Implantable middle ear hearing devices such as Vibrant Soundbridge have been used as an alternative to conventional hearing aids for the rehabilitation of sensorineural hearing loss. Other than the obvious disadvantage of requiring implantation middle ear surgery, it also applies a direct weight on the ossicular chain which, in turn, may have an impact on residual hearing. Previous studies have shown that applying a mass directly on the ossicular chain has a damping effect on its response to sound. However, little has been done to investigate the magnitude and the frequency characteristics of the mass loading effect in devices such as the Vibrant Soundbridge. METHODS: Five fresh cadaver temporal bones were used. The stapes displacement was measured using laser Doppler vibrometry before and after the placement of a Vibrant Soundbridge floating mass transducer. The effects of mass and attachment site were compared with the unloaded response. Measurements were obtained at frequencies between 0.1 and 10 kHz and at acoustic input levels of 100 dB sound pressure level. Each temporal bone acted as its own control. RESULTS: Placement of the floating mass transducer caused a reduction of the stapes displacement. There were variations between the bones. The change of the stapes displacement varied from 0 dB to 28 dB. The effect was more prominent at frequencies above 1,000 Hz. Placing the floating mass transducer close to the incudostapedial joint reduced the mass loading effect. CONCLUSION: The floating mass transducer produces a measurable reduction of the stapes displacement in the temporal bone model. The effect is more prominent at high frequencies. PMID- 15793407 TI - Experience with the SOUNDTEC implantable hearing aid. AB - OBJECTIVE: To assess the efficacy, morbidity, and patient satisfaction of the SOUNDTEC semi-implantable hearing aid. STUDY DESIGN: Retrospective case review. SETTING: Two tertiary referral centers (the Silverstein Institute and the Atkins Institute). PATIENTS: Sixty-four (four bilateral placements) patients with bilateral moderately severe sensorineural hearing loss: 47 patients had previously worn hearing aids. INTERVENTIONS: After separating the incudostapedial joint, a magnet encased in a titanium canister with a ring was introduced onto the stapes neck. Gelfoam or adipose tissue was used to stabilize the magnet. After 3 months, the external processor was fitted. MAIN OUTCOME MEASURES: At 1 month, audiometric testing was performed and functional gain was assessed. Patient acceptance and implant performance were measured by a visual analogue questionnaire. RESULTS: The device produced an average functional gain of 26 dB. Fifty-five percent of patients complained of magnet movement; this was eliminated in 80% of Silverstein Institute patients when the external processor was worn and fat was used to stabilize the magnet. CONCLUSION: The SOUNDTEC direct device is well tolerated in the majority of patients, with a significant increase in functional gain. Magnet instability and noise were the most frequent complaints and improved with processor placement and anchoring the magnet with fat. This electromagnetic semi-implantable hearing aid confers greater functional gain over conventional hearing aids and reduces occlusion effect and feedback. PMID- 15793409 TI - Ferromagnetic movements of middle ear implants and stapes prostheses in a 3-T magnetic resonance field. AB - HYPOTHESIS: A 3-T magnetic resonance field may cause motion or displacement of middle ear implants not seen in studies with 1.5-T magnets. BACKGROUND: Previous publications have described the safety limitations of some otologic implants in 1.5-T magnetic resonance fields. Several company-wide recalls of implants were issued. No studies to date have been reported for otologic implants within a 3-T magnetic resonance field, nor have there been comparisons with a 1.5-T field strength. METHODS: Eighteen commonly used middle ear implants and prostheses were selected. In Part 1, the prostheses were placed in Petri dishes and exposed to a 3-T magnetic resonance field. In Part 2, the particular prostheses that showed movement in Part 1 were placed into their intended use positions within temporal bone laboratory specimens and exposed to a 3-T field. Both parts were repeated in a 1.5-T field. RESULTS: In Part 1, three prostheses moved dramatically from their start positions when exposed to the 3-T magnetic resonance field. In Part 2, the three particular prostheses that showed movement in Part 1 showed no gross displacement or movement from their start positions within the temporal bone laboratory specimens. No implants moved in the 1.5-T field in either Part 1 or Part 2. CONCLUSION: Certain stapes prostheses move dramatically in Petri dishes in 3-T fields. When placed into temporal bone laboratory specimens, the same prostheses show no signs of movement from the surgical site in a 3-T field, and it appears that the surgical position holds the implants firmly in place. Results of published 1.5-T field studies should not be used directly for safety recommendations in a 3-T magnetic resonance. Heat, voltage induction, and vibration during exposure to the magnetic resonance fields should be considered as additional possible safety issues. Preference should be given to platinum and titanium implants in manufacturing processes and surgical selection. PMID- 15793410 TI - Neurotologic skull base surgery in pediatric patients. AB - OBJECTIVE: Innovations in diagnosis, surgical techniques, and perioperative care have dramatically improved outcomes in lateral skull base procedures in recent years. There is a belief, however, that children with skull base tumors have yet to benefit from these technological and procedural advances. The purpose of this study is to provide a clinical review of neurotologic skull base surgery in the pediatric population. STUDY DESIGN: Retrospective case review. SETTING: Private practice tertiary referral center. PATIENTS: Eighty-nine pediatric patients undergoing 115 neurotologic procedures for lateral skull base tumors from July 1992 to September 2003. MAIN OUTCOME MEASURES: Initial clinical presentation, tumor type, pre- and postoperative hearing and facial nerve status, treatment course, complications, and functional outcomes. RESULTS: The majority of tumors in this series were vestibular schwannomas, and 65 patients were diagnosed with neurofibromatosis Type 2. Surgical approaches included 70 middle fossa, 40 translabyrinthine, 2 transcochlear, 2 infratemporal fossa, and 1 retrosigmoid craniotomy. Complete tumor removal was accomplished in the majority of cases (97%), with good preservation of facial nerve function (House-Brackmann Grade I or II) in 80% of patients. In patients undergoing middle fossa surgery for hearing preservation, measurable hearing was preserved in 61.4% of cases. The incidence of complications was low. CONCLUSION: With advances in diagnostic procedures and use of current neurotologic techniques, pediatric patients may undergo successful treatment of lateral skull base tumors, with good functional outcomes and minimal morbidity. PMID- 15793411 TI - Hereditary bilateral conductive hearing loss caused by total loss of ossicles: a report of familial expansile osteolysis. AB - OBJECTIVE: The objective of this study was to report on three members of a family with familial expansile osteolysis; the important point about these patients was that none of them had middle-ear ossicles. STUDY DESIGN AND SUBJECTS: A retrospective case review including three cases with familial expansile osteolysis. SETTING: Department of Otolaryngology in a tertiary referral center. INTERVENTIONS: Each patient underwent computerized tomography of the temporal bone in the coronal view, audiometric and tympanometric evaluations, biochemical investigation, whole body isotope scans by Tc-99 mMDP and X-ray. Also the patients' pedigree was studied. Two of the patients had exploratory middle-ear surgery as well. RESULTS: The temporal-bone computed-tomography scan in the coronal view of all three patients and also exploratory middle-ear surgery, which was done on two of the patients, showed no ossicles in the middle ear of either ear in all three cases. This feature hadn't been reported in previous studies. Hearing loss was revealed in the medical histories since childhood. Audiometry indicated mild to moderate conductive and mixed hearing loss and also an AD-type tympanogram pattern along with an absence of acoustic reflexes in both ears of the cases. Both serum alkaline phosphatase and hydroxyproline levels were elevated. There was an increase in uptake and activity at multiple foci of the whole skeleton. No improvement in hearing thresholds was obtained after reconstruction of the middle ear. CONCLUSION: The total absence of middle-ear ossicles can probably be regarded as a new symptom in some patients with familial expansile osteolysis. Common ossiculoplasty for improving the hearing thresholds in this condition may be unsuccessful; therefore, both surgeons and patients must be completely aware of the contingent undesirable results. PMID- 15793412 TI - Sudden sensorineural hearing loss associated with inner ear anomaly. AB - OBJECTIVE: This study was conducted to evaluate the frequency of inner ear anomaly in patients with sudden sensorineural hearing loss and in control subjects. STUDY DESIGN: Retrospective case review. SETTING: A tertiary referral center. PATIENTS AND INTERVENTION: We evaluated 366 patients (165 men and 201 women; age range, 3-91 yr) with sudden sensorineural hearing loss and 228 control subjects without sensorineural hearing loss using magnetic resonance imaging. Three hundred fifty-six patients had unilateral and 10 patients had bilateral sudden sensorineural hearing loss. RESULTS: Eleven (2.9%) of 376 ears with sudden sensorineural hearing loss had inner ear anomaly. Nine patients (2.5%) had inner ear anomaly associated with sudden sensorineural hearing loss, but none of the 228 control subjects had the anomaly. The current study demonstrated that the frequency of inner ear anomaly in patients with sudden sensorineural hearing loss was significantly higher than in control subjects. CONCLUSION: Our study reveals that inner ear anomaly may be associated with sudden sensorineural hearing loss in 2.5% of patients. PMID- 15793413 TI - Inlay butterfly cartilage tympanoplasty in children. AB - OBJECTIVE: To assess the results of inlay butterfly cartilage tympanoplasty in children. STUDY DESIGN: Before-and-after trial; follow-up duration, 26.6 +/- 19.9 months (mean +/- standard deviation). SETTING: Tertiary referral center. PATIENTS: Fifty-nine pediatric cases of tympanic membrane perforation. INTERVENTION: Inlay butterfly cartilage tympanoplasty was performed under general anesthesia according to the technique originally described by Eavey and modified by Lubianca-Neto (i.e., without any associated split-thickness skin graft). MAIN OUTCOME MEASURES: Percentage of perforation closures, surgical complications, preoperative and postoperative puretone hearing thresholds; the results of inlay butterfly cartilage tympanoplasty were compared with those obtained in a retrospective series of 29 underlay fascia temporalis myringoplasties. RESULTS: The 71% "take rate" of inlay butterfly cartilage tympanoplasty was not significantly different from the 83% take rate obtained with underlay fascia temporalis tympanoplasty (p = 0.23, chi test). The anatomic results were improved when the graft diameter was at least 2 mm larger than the size of the perforation (81% take rate) (p = 0.009, chi test). No iatrogenic cholesteatoma was observed. Pure-tone hearing thresholds were improved at 0.5, 1, and 2 kHz, and stable at 4 kHz. Hearing levels were not different from those obtained with underlay fascia temporalis tympanoplasty. CONCLUSION: Inlay butterfly cartilage tympanoplasty is a safe, efficient, time-saving, and easy technique of tympanoplasty in children. Anatomic results may be improved by associating a split-thickness skin graft and/or by trimming a tragal graft much larger than the size of the perforation. PMID- 15793414 TI - Accuracy of tympanometric middle ear pressure determination: the role of direction and rate of pressure change with a fast, modern tympanometer. AB - HYPOTHESIS: Modern tympanometers run at higher rates of pressure change than older tympanometers, which increases the inaccuracy of determining the middle ear pressure. BACKGROUND: Tympanometric middle ear pressure may be susceptible to both the direction as well as the rate of pressure change, which is reflected by two different pressure peaks in a bidirectional recording. The resulting peak pressure difference results in an inaccuracy, which can amount to 25 daPa in older instruments with slower rates of pressure change. However, modern instruments often apply much faster rates, which may increase the peak pressure difference and thus the inaccuracy of middle ear pressure. METHODS: Middle ear pressure was measured for a negative and positive direction of pressure change at four different rates (50, 100, 200, and 400 daPa/s) in 38 normal adults. The peak pressure difference was calculated by the middle ear pressure determined in positive minus negative direction. RESULTS: The mean peak pressure differences ranged between 10 and 12 daPa (standard deviation = 8-11) in the four groups and were independent of the rate of pressure change (p = 0.321). CONCLUSION: The peak pressure differences found by the current tympanometer were consistently small for all rates of pressure change and were thus independent of the rate. This means that high rates can be used without decreasing accuracy, and the mean error is only 5 to 6 daPa, corresponding to the intrinsic hysteresis of the middle ear system. PMID- 15793415 TI - Repositioning maneuver for the treatment of the apogeotropic variant of horizontal canal benign paroxysmal positional vertigo. AB - OBJECTIVE: The purpose of this study was to determine the effectiveness of a new physical maneuver in the treatment of the apogeotropic variant of horizontal canal benign paroxysmal positional vertigo. STUDY DESIGN: Case review. SETTING: Outpatient clinic. PATIENTS: The diagnosis of apogeotropic horizontal canal benign paroxysmal positional vertigo was based on the history of recurrent sudden crisis of vertigo associated with bursts of horizontal apogeotropic paroxysmal nystagmus provoked by turning the head from the supine to either lateral position. The patients were three men and five women ranging in age from 31 to 73 years (average, 49.2 yr). INTERVENTIONS: All patients were treated with a repositioning maneuver based on the hypothesis that the syndrome is caused by the presence of free-floating dense particles inside the endolymph of the anterior arm of the horizontal canal. The maneuver favors their shifting into the posterior arm of the canal. Patients were reexamined immediately after the treatment and underwent Gufoni's liberatory maneuver for the geotropic variant of horizontal canal benign paroxysmal positional vertigo. MAIN OUTCOME MEASURE: The treatment outcome was considered as responsive when, after one repositioning maneuver, nystagmus shifted from apogeotropic to geotropic. RESULTS: The repositioning maneuver resulted in a transformation from the apogeotropic variant into a geotropic variant of horizontal canal benign paroxysmal positional vertigo in all patients. CONCLUSION: This maneuver represents a simple and effective approach to the treatment of the apogeotropic variant of horizontal canal benign paroxysmal positional vertigo. It favors the shifting of the canaliths from the anterior into the posterior arm of the horizontal canal from where they can migrate into the utricle with Gufoni's maneuver. PMID- 15793416 TI - Vestibular schwannoma in the only hearing ear: cochlear implant or auditory brainstem implant? AB - OBJECTIVE: To explore the dilemma faced by neurotologists confronted with the patient who develops a vestibular schwannoma in the only hearing ear, the other having been deaf from birth, and to consider the choice between auditory rehabilitation using a cochlear implant (CI) on the congenitally deaf side and an auditory brainstem implant (ABI) on the tumor side. STUDY DESIGN: A record review of two patients born deaf in one ear and who developed a vestibular schwannoma in the contra lateral ear, who then received a CI in the congenitally deaf ear. SETTING: Tertiary referral center with special experience in vestibular schwannoma surgery, neurofibromatosis type 2 management, and cochlear implantation. RESULTS: Neither patient was a good CI user. At 1 year postimplant, they both scored 0% on abbreviated words and 0% and 7%, respectively, on Bench Kowal Bamford sentences. They scored 54% and 57%, respectively, on City University of New York sentences with lip-reading, and both had fair access to environmental sound scoring at 45% each. CONCLUSIONS: The results from cochlear implantation in the congenitally deaf ear in these patients were poor and suggest that stimulus deprivation in the early stages of the maturation of the auditory pathways is important even for a unilateral hearing loss. Unfortunately, the factors that predict a good ABI result are not known. In these circumstances, the authors advocate the insertion of an ABI at the time of tumor removal, retaining the option of CI in the congenitally deaf ear in the event of a poor outcome with the ABI. PMID- 15793417 TI - Effects of vestibulo-ocular reflex exercises on vestibular compensation after vestibular schwannoma surgery. AB - OBJECTIVE: To assess vestibular function in a large group of vestibular schwannoma patients so that we could determine whether simple vestibular exercises speed vestibular dysfunction recovery after tumor removal surgery. STUDY DESIGN: A prospective investigation of the vestibular dysfunction experienced by patients in the first 12 weeks after surgery. SETTING: Vestibular investigation unit at a tertiary referral institution. PATIENTS: Sixty-five patients with identified vestibular schwannoma referred for preoperative vestibular investigations. Thirty-two men and 33 women, with a mean age 51 years (range, 24-77 yr). INTERVENTIONS: There were 27 control patients, 30 exercise patients, and 8 patients that had balance physiotherapy. Exercise patients began simple vestibulo-ocular reflex gaze stabilization exercises 3 days after surgery. MAIN OUTCOME MEASURES: Postoperative vestibular function testing was performed at 2 to 3, 6 to 7, and 10 to 12 weeks after surgery. Objective measurements of vestibular compensation status were as follows: spontaneous nystagmus and sinusoidal harmonic acceleration asymmetry and gain values. Dizziness Handicap Inventory questionnaires were used to assess subjective perceptions. RESULTS: The main findings were reduced dispersion in vestibulo-ocular reflex asymmetry at 2 to 3 weeks, reduced mean in asymmetry at 6 to 7 weeks, less dizziness/imbalance according to the Dizziness Handicap Inventory questionnaire, and that preoperative caloric tests did not predict postoperative severity of vestibular systems. CONCLUSION: This large study provided unique evidence that a program of simple vestibular exercises and education can speed the rate of compensation after vestibular schwannoma surgery. PMID- 15793418 TI - Prediction of long-term facial nerve outcomes with intraoperative nerve monitoring. AB - OBJECTIVE: Evaluate the utility of a statistical equation using two independent intraoperative monitoring parameters in predicting long-term facial nerve function. STUDY DESIGN: Retrospective case review. SETTING: Tertiary care hospital. PATIENTS: Sixty patients undergoing resection of vestibular schwannomas with intraoperative facial nerve monitoring at a single institution. INTERVENTION: All patients underwent microsurgical resection of vestibular schwannomas with the use of intraoperative cranial nerve monitoring. MAIN OUTCOME MEASURE: Final facial nerve outcome measured using the House-Brackmann scale at least 6 months after microsurgical resection. RESULTS: Five out of 60 (8.3%) patients demonstrated significant long-term weakness (i.e., House-Brackmann grade III or worse). Intraoperative monitoring parameters (proximal stimulation threshold, proximal-to-distal response amplitude ratio) were accurate in predicting increased risk of long-term facial nerve dysfunction when used in a logistic regression model. A Student's t test confirmed the equation result was statistically significant in differentiating long-term facial nerve outcomes. CONCLUSION: Patients with immediate weakness are at higher risk of having long term poor facial nerve function. The use of intraoperative monitoring parameters was reliable in predicting facial nerve outcomes. Patients with permanent facial nerve dysfunction often require rehabilitative procedures. The ability to predict facial nerve outcomes with intraoperative monitoring may allow early rehabilitative procedures to improve quality of life and prevent ocular complications. PMID- 15793419 TI - Intraoperative electrophysiologic identification of the nervus intermedius. AB - OBJECTIVE: Although enormous attention has been directed to the localization and preservation of the facial nerve in acoustic neuroma surgery, the nervus intermedius has largely been ignored. In this article, we describe a method for intraoperative electrophysiologic identification of the nervus intermedius. STUDY DESIGN: Retrospective case review. SETTING: University hospital (tertiary care center). PATIENTS: Thirty-three patients who underwent intraoperative facial nerve monitoring for various cerebellopontine angle procedures. Recording electrodes were placed in the orbicularis oculi and orbicularis oris muscles. A constant-voltage stimulator was used to stimulate both the facial nerve and the nervus intermedius. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Electrophysiologic response after stimulation of the nervus intermedius. RESULTS: Stimulation of the nervus intermedius produced long-latency, low-amplitude response recorded only on the orbicularis oris channel. The response had a mean threshold 0.4 V, a mean latency of 11.1 ms, and a mean amplitude of 11.1 microV, all significantly different from responses to stimulation the facial nerve. CONCLUSION: Knowledge of electrophysiologic features of nervus intermedius stimulation can help protect the facial nerve during cerebellopontine angle surgery. The surgeon must recognize that stimulation of the nervus intermedius can cause electromyographic activity in the facial nerve monitoring channels, but the main trunk of the facial nerve may lie in entirely different location in the cerebellopontine angle. PMID- 15793420 TI - Objective assessment of the perineural vascular plexus as a landmark for the horizontal part of the facial nerve in middle ear and mastoid surgery. AB - OBJECTIVE: To objectively evaluate the usefulness and the reliability of the perineural vascular plexus as a landmark for identification of the facial nerve in surgery for chronic squamous otitis media. STUDY DESIGN: Prospective case series. SETTING: Tertiary otologic center. PATIENTS: Seventy consecutive patients requiring surgery for cholesteatomatous otitis media. INTERVENTION: Use of a semiquantitative grading system intraoperatively to assess the utility and ease of using the perineural facial plexus as the pointer to the facial nerve. MAIN OUTCOME MEASURES: Description of the perineural vascular plexus and assessment of the reproducibility of the grading system. RESULTS: In 82.5% of patients, the plexus was used as the sole pointer to the level of the facial nerve, with other landmarks being used in the remaining 17.5%. A very prominent vessel was used to identify the nerve in 82.5%, therefore being classified as Grade 1. Multiple small vessels were seen in 15.8% (Grade 2), and in 1.5% the vessel plexus was thin and difficult to identify. The average measure intraclass correlation was 0.75 (95% confidence interval, 0.57-0.85), indicating excellent reproducibility of the system. CONCLUSION: We believe that the perineural vascular plexus is a dependable and reliable landmark for the level of the facial nerve in surgery for chronic otitis media. PMID- 15793421 TI - A comparison of surgical techniques used in dynamic reanimation of the paralyzed face. AB - OBJECTIVES: To compare the outcomes of three surgical techniques used in the rehabilitation of the paralyzed face. STUDY DESIGN: Retrospective study. SETTING: University-based tertiary referral center. PATIENTS: Between 1976 and 2000, rehabilitative facial nerve surgery was performed on 70 adult patients with varying underlying diseases. INTERVENTION: Three methods of facial nerve rehabilitative surgery were performed: end-to-end anastomosis, cable nerve graft interposition, and classic faciohypoglossal transposition. MAIN OUTCOME MEASURES: The House-Brackmann grade was scored at 6, 12, 24, and 36 months by the two senior authors. A favorable outcome was defined as House-Brackmann Grade 59 years) age groups. Sagittal oblique images were obtained at 1-mm intervals through the midface, perpendicular to the nasolabial fold. Quantitative analyses of the cheek fat pad overlying the levator labii superioris and zygomaticus major muscles were performed separately. RESULTS: For both mimetic muscles, there were no significant differences between young and old subjects in muscle length, thickness, or volume from muscle origin to nasolabial fold. In addition, there were no significant differences between age groups in fatty infiltration of the muscles. The volumes of the medial and lateral aspects of the cheek fat pad were significantly greater in old than in young subjects (p < 0.05). In young subjects the greatest distribution of fat pad volume was found in the middle third of the cheek mass. There was a significant reduction in that portion of the fat pad distributed in the upper third and a further reduction in the lower third (p < 0.01). In old subjects, because of the increased distribution of fat in the upper third of the cheek fat pad, there was no significant difference in volume between the upper and middle thirds of the cheek fat pad. The percentage increase in fat in the upper third relative to the remainder of the midface was significantly greater in old compared with young subjects (p < 0.01). In contrast, the volume of the lower third of the midface was not significantly different between young and old subjects. CONCLUSIONS: The present study indicates that ptosis alone does not account for the changes observed in the aging midface. Selective hypertrophy of the upper portion of the cheek fat pad was also observed. The mimetic muscles, on the other hand, showed no significant differences with aging. To attain maximum precision in facial rejuvenation, these data suggest that after suspension of the ptotic cheek fat pad, each patient should be evaluated for excess bulk in the upper portion of the cheek fat pad. If excess bulk is present, patients may benefit further from selective reduction directed to the upper portion of the cheek fat pad, remaining superficial to the mimetic muscles of the face. PMID- 15793459 TI - A prospective study of anti-aging topical therapies using a quantitative method of assessment. AB - BACKGROUND: In the treatment of photoaged skin, glycolic acid works by removing superficial portions of the epidermis and stimulating dermis regeneration. Vitamins A, C, and E should stimulate collagen production and antioxidants should prevent free radical damage and skin aging. However, the effectiveness of different therapies has often relied on subjective methods of assessment. Histologic analysis has seldom been used because of the drawback of permanent scarring. In the literature, the use of a quantitative method for the assessment of facial rejuvenation has been described: the silicone replica technique. The authors' aim was to promote and recommend the use of this technique and, in particular, to test the effect of glycolic acid and multivitamin- and antioxidant based products on skin texture. METHODS: The authors performed a prospective, randomized, double-blind, controlled study on 30 women treated topically in the outer canthal region (crow's-feet area). Patients were divided into three groups (groups A, B, and C); each group consisted of five patients between the ages of 31 and 40 years and five patients between the ages of 41 and 50 years. Group A was treated by glycolic acid application, initially at home for 2 weeks, followed by a higher concentration administered in the office weekly for six applications. Group B was treated by topical application at home of a multivitamin product daily for 3 months. Group C was treated with a cream base (placebo) for 3 months and represented the control group. Skin areas under treatment were photographed and reproduced by the silicone replica technique at baseline and at the end of treatment. This technique reproduces exactly the skin's texture. Digital images were obtained from skin replicas and analyzed by specific software for different parameters: roughness, microsulcus number, and width. Pretreatment and posttreatment values were compared using the Wilcoxon signed-rank test. RESULTS: In group A, microsulcus number and width were statistically decreased, but roughness was not. In groups B and C, parameters were not statistically modified. CONCLUSIONS: The silicone replica technique allowed a quantitative analysis of results obtained with different topical therapies. In particular, it confirmed the efficacy of glycolic acid in skin rejuvenation. PMID- 15793462 TI - Endoscopic correction of eyebrow asymmetry in associated forehead soft-tissue injuries. AB - Supported by anatomical principles, a subgaleal endoscopically assisted selective myotomy of the forehead and glabellar muscles was used to achieve eyebrow symmetry in patients with unilateral facial nerve palsy and associated forehead soft-tissue injuries. Selective myotomy equalizes the agonist-antagonist muscle relationship on both sides of the forehead, thereby allowing the untouched muscle to exert its activity without opponent restriction. Selective myotomy allows for the management of the muscles' group action as well as individual manipulation of each muscle in accordance with its participation in the eyebrow deformity. PMID- 15793461 TI - The "pixie" ear deformity following face lift surgery revisited. AB - BACKGROUND: The "pixie" ear deformity can be recognized by its "stuck on" or "pulled" appearance, which is caused by the extrinsic pull of the medial cheek and jawline skin flaps at the earlobe attachment point, the otobasion inferius. The tension results in migration of the otobasion inferius from a posterior cephalad position to an anterior caudal position. Although this deformity has been described clinically, it has yet to be objectively defined. METHODS: Recently, the two components of the earlobe, the attached cephalic segment (intertragal to otobasion inferius distance) and the free caudal segment (otobasion inferius to subaurale distance), were shown to be essential in evaluating for earlobe ptosis and pseudoptosis. These two components can be used to designate an objective criterion for the pixie ear deformity. The deformity, as defined by the authors' parameters, was assessed in 44 patients who had undergone rhytidectomy. A simple and accurate surgical treatment is demonstrated by a cadaver dissection and a clinical case. RESULTS: The deformity can be defined as an increase in the attached cephalic segment (intertragal to otobasion inferius distance) and a decrease in the free caudal segment (otobasion inferius to subaurale distance) to 0 mm following rhytidectomy. The incidence of pixie ear deformity was 5.7 percent in the authors' series of patients. CONCLUSIONS: A medially based triangular excision over the attached cephalic segment is presented as a simple and accurate surgical treatment of pixie ear deformity. A more accurate and objective designation may allow for improved detection, avoidance, and treatment of this deformity. PMID- 15793463 TI - Vertical mammaplasty. AB - Current criticisms regarding vertical mammaplasty include problems with poor immediate postoperative appearance, nipple-areola complex malposition, and excessive lower pole length. These problems can be avoided by proper patient selection, by utilizing correct concepts of skin design, and by observing correct glandular resection and closure concepts. Vertical mammaplasty also can result in other problems, such as hypertrophic circumareolar scars and lower pole deformities, including notching, boxy shape, infra-areolar depression, and flatness. These problems are also largely avoidable by using correct technique. Several basic concepts described previously have not proven necessary to achieve good results. Abandoning some of these principles has contributed to the ability to establish an aesthetically ideal breast shape intraoperatively as well as to a decrease in morbidity. This includes eliminating liposuction as a major integral component of the procedure, eliminating suturing the gland to the pectoralis muscle, not undermining the lower pole skin, and avoiding overly wide skin resection and tight wound closure that produces significant lower pole distortion in the early postoperative period. An important concept that has proven reliable is to use a "closed" design that does not predetermine the areolar opening whenever circumstances permit. When this is not possible, a modification that utilizes the smallest possible circumference as an open design is better than a large "mosque." These alternatives allow greater flexibility in determining final nipple position and also reduce the risk of hypertrophic circumareolar scars. Important glandular resection concepts include creating pillars that are attached to both the skin and the chest wall; making them of adequate dimension to avoid postoperative lower pole shape problems, such as flattening; resecting closer to the skin lateral to the pillars to avoid a boxy breast shape; and using a drain both to assist in accurately determining the endpoint of resection and to avoid postoperative seromas. Key closure concepts include approximation of the superior surfaces of the pillars at their base to maintain vertical height and thereby prevent lower pole flattening; approximation of the inferior surfaces of the pillars to the base of the breast to prevent notching; and proper management of the vertical incision by restricting the purse-string suture effect to only the inferior portion of the incision, where there may be skin excess present. Inclusion of these concepts leads to predictable and improved aesthetic results in vertical mammaplasty. This allows full realization of the purported advantages of vertical mammaplasty and allows this method to be utilized with a level of confidence similar to that seen with inverted-T techniques. PMID- 15793466 TI - Brody's article on "the perfect breast". PMID- 15793467 TI - Brody's article on "the perfect breast". PMID- 15793468 TI - Brody's article on "the perfect breast". PMID- 15793469 TI - Brody's article on "the perfect breast". PMID- 15793471 TI - A simple device for cleaning the hair after scalp surgery. PMID- 15793472 TI - Rate of occurrence of dexamethasone-induced cleft palate affected by uterine environment in the mouse. PMID- 15793473 TI - Ambulatory cleft lip and palate surgery by tumescent technique. PMID- 15793474 TI - Cleft palate and congenital alveolar synechiae syndrome. PMID- 15793475 TI - Median palatine cyst: report of an unusual entity. PMID- 15793476 TI - TRAM flap after laparoscopic hernia repair. PMID- 15793477 TI - The posterior interosseous nerve as a donor source for nerve grafting: a concept revisited. PMID- 15793478 TI - A very severe hand pathology with a possible diagnosis of onychogryposis. PMID- 15793479 TI - Repair of fingertip amputations with local digital flaps and perionychial composite grafts from the amputated part. PMID- 15793480 TI - Vascular leiomyoma of the hand. PMID- 15793482 TI - A tendon approximator. PMID- 15793483 TI - Management of deep palmar burns with reverse prefabricated radial fascial flaps. PMID- 15793484 TI - Successful split-thickness skin grafting in a contaminated wound with an enterocutaneous fistula. PMID- 15793485 TI - Estimation of volumetric indices of skin microvascular blood flow with laser Doppler flowmetry. PMID- 15793486 TI - Congenital hypertrophy of the abductor digiti minimi muscle of the foot. PMID- 15793487 TI - Thoracic aortic endografting. PMID- 15793488 TI - Traumatic aortic ruptures. AB - Most patients with a traumatic aortic rupture (TAR) were involved in high velocity motor vehicle accidents. Initial management of these patients should be according to the guidelines of the advanced trauma life support group (ATLS). Patients with a suspected TAR are preferably managed by controlled hypotension. TAR can be diagnosed by spiral CT angiography, additional angiography is not needed. Although results from conventional surgery have improved over the years, results from endovascular grafting are better with reduced mortality and paraplegia rates. Acute open surgery has become the second choice and patients are preferably managed by endovascular treatment. The endovascular graft can be placed with a minimal invasive procedure and delay in treatment because of accompanying injuries is avoided. Patients with a TAR treated by an endovascular graft should be kept under surveillance. For optimal patient care level I trauma centers should have thoracic endovascular grafts available for direct use. PMID- 15793489 TI - Endograft repair of acute aortic dissection. Promises and challenges. AB - Acute dissection is a uniquely complex, relatively common, and frequently lethal aortic catastrophe. Historically, surgical treatment has been reserved for cases with complications including rupture; the results have been less than optimal because of excessive morbidity and mortality. This is the main reason why conservative management emerged as the standard of care for management of acute type B aortic dissection (TBAD). While more patients would appear to survive with a conservative treatment strategy, the outcome in terms of 30-day mortality (20%) and occurrence of late complications - such as enlarging aneurysms of the thoracic false lumen (30-40%) - leaves (again) much to be desired. Stent-graft endovascular repair has emerged as a very promising, less invasive treatment option. These devices, when used appropriately, can achieve the important therapeutic goals of entry-site coverage, depressurization of the false lumen, and expansion of the compressed true lumen - overcoming ischemic (malperfusion) manifestations. The early results of stent-graft repair of TBAD are encouraging, and even exciting, but much more work needs to be done in various critical areas surrounding this condition. Thoracic endograft technology has lagged behind its abdominal counterpart. The design of acute dissection-specific devices is imperative, reflecting the significant differences between TBAD and degenerative thoracic aortic aneurysm disease. Needs for this and other important developments notwithstanding, it is generally acknowledged that stent-graft intervention does represent an important advance in the treatment of patients with TBAD. Well designed, controlled clinical trials will be necessary to elucidate the relative value of several endovascular thoracic strategies. PMID- 15793490 TI - Device-specific outcomes with endografts for thoracic aortic aneurysms. AB - For the endovascular therapy of abdominal aortic aneurysm an increasing number of publications deal with the subject of device-related outcome and aneurysm sac behaviour. The present study was designed to investigate whether, and to what extent, aneurysm sac behaviour of thoracic aortic aneurysms and perioperative outcome is influenced by the different types of stent grafts. Bibliographic search and analysis was performed. A systematic MEDLINE search on thoracic aortic aneurysms produced a total of 2,884 references with abstracts between 1990 and 2004. Restricted by using the additional terms of ''descending'' and ''stent graft'' the result condensed to 152 papers. Each reference was analyzed according to predetermined criteria. The majority of articles were published in the English language. Two publications summarized the results of nation-wide or continental registers of endoluminally treated patients with descending aortic dissections or thoracic aortic disease. Several series were partial and/or updated reports from the same authors and/or on the same patients. A total of 48 centres reported about the endovascular treatment of more than 3,000 patients with pathological thoracic aortic entities (Approximately 1,650 degenerative thoracic aortic aneurysms, Approximately 1,000 aortic dissections, Approximately 400 miscellaneous aortic pathologies). The heterogeneity among the series precluded any relevant comparison. Only 4 centres have presented study groups of more than 100 treated patients, but 3 series contained several treated aortic pathologies. Without a differentiation to the underlying pathology, the bibliographic research documented a rate of technical success in a range of 76% to 100%. Summarizing the data of par-procedural and in-hospital mortality the bibliographic research documented a mortality rate of 6.8% for all reported procedures. The rate of paraplegia for all procedures was 1.7%. In conclusion the presented bibliographic search and analysis demonstrated the technical feasibility of the endoluminal stent grafting of a great variety of thoracic aortic pathologies. The short-term results are promising, but data of mid-term results are rare and long-term results are missing. Moreover, the question about the influence of different types of stent grafts on the outcome can not be answered. PMID- 15793491 TI - Stentgrafting of the thoracic aorta-complications. AB - Endovascular therapy (EVT) of thoracic aortic pathologies meanwhile is an established procedure with favourable midterm results in high risk patients. Different stent fabrications are available with defined flexibility, radial attachment force, metallic stent components and membrane porosities. Recent approval of the TAG Excluder (Gore) by the Food and Drug Administration (FDA) was an important step. Endoleaks, paraplegia, stroke and retrograde dissections are the main specific complications. Type I endoleak incidence rates are related to morphological case complexity; primary frequency rates of 0-20% are reported in the literature with 0-5% secondary incidence. Creating an appropriate proximal neck -- if necessary by supra-aortic branch remodelling -- and deliberate left subclavian artery overstenting is the key mechanism to avoid proximal endoleaks. Paraplegia rates are reportedly low with EVT in the range of 0-5%. Risk situations are: cases of rupture with compromised blood pressure, cases with a history of abdominal aortic aneurysm (AAA) exclusion, cases who require total overstenting of the descending thoracic aorta. The role of cerebrospinal fluid (CSF) drainage in EVT is not defined. Stroke as consequence of embolizing material from central endoaortic manipulations is almost in the same range as paraplegia, when morphologies in the distal arch are attacked. Retrograde dissection is reported not only after treatment of type B dissection but also after aneurysms. Rigid bare springs and ballooning in cases of type B dissection seem to be involved. In recent reports mortality in elective cases varies between 1.5% and 6.5%. All these data are promising but the proof of longterm durability is still lacking. Further development will show whether or not these preliminary data will translate into longterm success. PMID- 15793492 TI - Fenestrated and branched endovascular techniques for thoraco-abdominal aneurysm repair. AB - Since 1991, endovascular aortic aneurysm repair (EVAR) has been established as an alternative for open surgical repair of aortic aneurysms. One of the main limitations for EVAR is the need for a sufficient sealing zone below or above vital aortic side branches. Recently, efforts have been made to overcome these limitations by incorporating fenestrations or branches in customized stent grafts. This paper reviews the technical and clinical possibilities, as well as the results with fenestrated and branched stent-grafts. All these techniques can be classified into 6 groups, including abdominal fenestrated, abdominal branched, thoraco-abdominal fenestrated, thoraco-abdominal branched, thoracic fenestrated, and thoracic branched stent-grafting. The only well-elaborated technique at this moment is abdominal fenestrated stent-grafting. Currently, branched stent grafting must be regarded as experimental, but advancements are taking place rapidly. It is anticipated that wider adoption will take place in the near future. New developments include the further use of reinforced fenestrations, indwelling catheters and flexor sheaths, as well as the use of new type branches. PMID- 15793494 TI - Stent-graft treatment of complete acute aortic transection complicated by intussusception and pseudo-coartaction. AB - Aim of the study was to validate the use of endoluminal stent-graft treatment as an alternative to conventional surgery in patients affected by blunt chest trauma and aortic disruption with multiple associated lesions. We report the case of a young female admitted with diagnosis of descending thoracic aortic transection and multiple traumas following a car accident. Spiral computed tomography revealed circular disruption of thoracic aorta immediately after isthmus region with intussusception of leaflets and pseudo-coartation. Doppler analysis showed a 70 mmHg transaortic gradient. The hemodynamic evaluation confirmed the existence of severe transaortic gradient. A Gore-TAG endoprosthesis (26 x 100 mm) was selected. Intraoper-ative transesophageal echocardiography assessment was performed to monitoring the entire procedure. The final arteriogram showed an optimal sealing at proximal and distal site without endoleak with complete readjustment of intimal leaflets to the aortic wall and disappearance of transaortic gradient related to the pseudo-coartation. No complication was observed in the early postoperative and patient was discharged one month later once complete rehabilitation of associated lesion was obtained. Computed tomography scan performed before discharge revealed persistency of patent lumen of aorta with fibrosis of readjusted circumferential intimal flap. In conclusion endovascular repair of complete aortic transection may result safe and effective particularly in patients with extensive associated injuries. Indeed the severity of coexisting non-aortic lesions could be adversely affected by conventional surgery in consideration of high surgical morbidity due to open thoracotomy. Stent-graft repair allows the patient to timely undergo medical or surgical management of associated lesions and a prompt rehabilitation with shorter hospital stay. PMID- 15793493 TI - Great vessels transposition and aortic arch exclusion. AB - AIM: We describe our experience in endovascular repair of Thoracic Aortic Aneurysms and Dissections (TAAD) involving the aortic arch in high risk patients (HRP). METHODS: Twenty-nine patients presented with TAAD involving the aortic arch and were treated by endovascular exclusion. Pathologies were as follows: atherosclerotic aneurysms of the descending thoracic aorta in 15 cases, acute Stanford type A dissections in 6 cases, Stanford type B dissections in 7 cases (1 acute), and 1 false aneurysm of the ascending aorta. Total-arch transpositions of all supra-aortic vessels (aortic debranching) to the ascending aorta were done in 11 cases throught median sternotomy. We performed carotido-carotid bypass (hemi arch transposition) in 16 patients by cervicotomy. Secondary to surgical transpositions, we placed endovascular stentgrafts in all but 2 patients for final exclusion, the 2 remaining being planned for later exclusion. The Talent, Excluder, TAG and Zenith endografts were used in 12, 3, 1 and 4 cases respectively. Banding technique was associated in some cases. RESULTS: All surgical transpositions were successful although 1 led to a minor stroke (1/29=3.5%), which worsened to major stroke after endovascular exclusion. Endovascular procedures were performed in all but one case (26/27=96.3%). Two patients (2/26=7.7%) died from catheterization related complications after endovascular exclusion (iliac rupture and left ventricle perforation). One patient had a delayed minor stroke (1/26=3.8%). Recirculation was found in 13.3% (2/15) of aneurysms and 27.3% of thoracic false channels. During a mean follow-up of 15.7 months (13 days to 45.5 months), 1 patient (1/26=3.8%) who had preoperative chronic pulmonary failure died at 6 months from respiratory worsening. We observed one case (3.8%) of unilateral limb palsy unrelated to cerebral ischemia, which we successfully treated by cerebrospinal fluid (CSF) drainage. No stent-related complication was seen. One new type 1 endoleak appeared at 12 months on an aneurysm, which resolved after stentgraft extension. Three thoracic dissection false channels remained patent during follow-up, of which one was retrograde originating distally in the descending aorta. CONCLUSIONS: Secondary endovascular exclusion of thoracic aortic diseases involving the arch in HRP is made feasible thanks to the preliminary aortic debranching. Total-arch transposition may be of greater interest in case of proximal neck length uncertainty and potential embolization from the aortic arch. Mid-term results are good although patients must be followed carefully to detect aortic recirculation and enlargement. PMID- 15793495 TI - Endovascular stent graft repair of mycotic aneurysms of the thoracic aorta. AB - Traditional open surgical repair for mycotic aneurysms of the thoracic aorta is associated with significant morbidity and mortality. Endovascular repair has been reported as an alternative treatment in patients with mycotic thoracic aneurysms. This article reports our experience of endovascular stent graft placement in three patients with mycotic aneurysms of the thoracic aorta and compares the results with similar reports on the literature. PMID- 15793496 TI - Retrievable inferior vena cava filters: early clinical experience. AB - AIM: Multiple-trauma patients often have injuries that prevent the use of anticoagulant or sequential compression device prophylaxis. Temporary inferior vena cava filters (IVCFs) offer protection against pulmonary embolism (PE) during the early, highest-risk perioperative and immediate injury period, while avoiding potential long-term sequelae of a permanent IVCF. The objective of this study was to evaluate the efficacy of prophylactic, temporary IVCF placement at the intensive care unit (ICU) bedside under real-time intravascular ultrasound (IVUS) guidance in multiple-trauma patients. METHODS: One hundred and three multiple trauma patients between July 1, 2002, and July 1, 2004, under-went placement of Gunther-Tulip (n=38), Recovery (n=30) or OptEase (n=35) retrievable IVCFs under real-time IVUS guidance. The mean+/-SD injury severity score of the patients was 27.7 (+/-2.2). All patients had abdominal X-rays to verify filter location. Before IVCF retrieval, all patients underwent femoral vein color-flow ultrasonography to rule out deep vein thrombosis (DVT) and pre and postprocedure vena-cavography for possible IVCF thrombus entrapment and postretrieval IVC injury. RESULTS: Twenty-four patients died of their injuries; no deaths were related to IVCF placement. One PE occurred during follow-up after filter retrieval, and 2 insertion site femoral vein DVT occurred. As verified by abdominal X-rays, 97.1% (100/103) of IVCFs were placed without complications at the L2-3 level. Filter-related complications included 3 groin hematomas (2.9%) and 3 IVCFs misplaced in the right iliac vein early in our experience; these filters were uneventfully retrieved and replaced in the IVC within 24 h. Forty four patients underwent uneventful retrieval of IVCFs after DVT or PE anticoagulation prophylaxis was initiated. Thirty-five filters were not removed, including 32 because severity of injury prevented DVT or PE prophylaxis and 3 because of thrombus trapped with the filter. CONCLUSIONS: Prophylactic, temporary IVCFs placed at the ICU bedside under IVUS guidance in multiple-trauma patients serves as an effective bridge to anticoagulation until venous thromboembolism prophylaxis can be initiated. Further investigation of this bedside technique and the role of temporary IVCFs in these patients is warranted. PMID- 15793497 TI - Elective popliteal aneurysms: does venous availability has an impact on indications? AB - AIM: The aim of this retrospective study was to evaluate the patency and limb salvage rates after prosthetics or venous bypasses for asymptomatic or mildly symptomatic popliteal aneurysms, in order to determine if small uncomplicated aneurysms (caliber <300%) should be operated or periodically controlled when a venous conduit is not available. METHODS: During a 18 years period, 100 popliteal aneurysms, including 85 asymptomatic and 15 associated with intermittent claudication, were operated on: group I consisted of 80 venous bypasses, and group II consisted of 20 prosthetic bypasses. RESULTS: Demographics and risks factors were similar in both groups. Local complications were more frequent in group I (17% vs 10% p=NS). Early vascular complications were rare in both groups. Late arterial complications were more frequent in group II. Primary patency, assisted primary patency, and secondary patency rates at 2 years were 94.3%, 97.3% and 98.7% in group I, and 61.5%, 89% and 88.4% in group II (p<0.05). In contrast, the limbs salvage rates were not significantly different for each type of graft (98.7% vs 100% p=NS). CONCLUSIONS: Our data shows that aneurysms treated with a prosthetic graft are at higher risk of late occlusion than those operated with a vein graft. This should be taken into account when facing a small uncomplicated aneurysm without available venous conduit. The presence of a suitable vein should be checked before deciding to operate a small uncomplicated popliteal aneurysm. PMID- 15793498 TI - Venous bypass of complicated stent placement for recurrent extracranial carotid occlusive disease. AB - Aim of the study was to to demonstrate a useful solution to carotid angioplasty and stent complications. A 67 year old male had uncomplicated left carotid endarterectomies in 1985 and 1986. A left distal common carotid angioplasty and stent in 1999 was complicated by stenosis. In 2000 a left common carotid bypass from the lower common carotid to the distal internal carotid well above the stent was performed, yielding retrograde filling of the external carotid, distal antegrade filling of the internal carotid, and widely patent vessels in subsequent Doppler studies. He is currently doing well clinically with no recurrent stenosis. In conclusion a second redo carotid operation was deferred in favor of angioplasty and stent, which had complications. Despite prior operations the only difficulty with the reoperation was obtaining control of the distal internal carotid above the stent. The strategy demonstrated here will be useful to correct complications of carotid angioplasty and stenting. PMID- 15793499 TI - Spontaneous iliac arteriovenous fistula: recognition and management. PMID- 15793500 TI - True aneurysm of an ilio-femoral saphenous vein graft. PMID- 15793501 TI - Rupture of a superficial femoral artery aneurysm following proximal and distal ligation and a by-pass procedure: a word of caution. PMID- 15793502 TI - A huge true aneurysm of the saphenous vein patch for anastomotic angioplasty: report of a case. PMID- 15793503 TI - Cutaneous lupus erythematosus induced by infliximab. PMID- 15793504 TI - Dermatoepidemiology. AB - Dermatoepidemiology is an important emerging discipline in dermatology. This article reviews clinical and analytic epidemiology pertinent to reading, interpreting, and critically examining the literature, and presents an overview of evidence-based dermatology as a starting point for further study. PMID- 15793505 TI - Adult-onset atopic dermatitis. AB - Adult-onset atopic dermatitis (AD) is a recently introduced subgroup of AD. Apart from the most typical flexural lichenified/exudative pattern in adults, patients may also have nontypical morphology and localization. The aim of this retrospective study was to find the frequency of nontypical morphology and localization in adult-onset AD and to evaluate the accuracy of United Kingdom Working Party's criteria in detecting those cases. Among 376 patients consecutively diagnosed with AD according to criteria of a previous study, 63 patients (34 women and 29 men) (16.8%) with onset of AD after the age of 18 years were allocated to the adult-onset group. A total of 7 patients (11.1%) had nonflexural involvement with nummular (6.3%), prurigo-like (3.2%), or follicular (1.6%) patterns that could not be attributed to contact sensitivities. A total of 14 patients (22.2%) did not fulfill the United Kingdom Working Party's criteria. It was interesting that United Kingdom Working Party's criteria did not cover the same patients as did the previous study's criteria in nearly one fourth of the cases. PMID- 15793506 TI - The effect of topical tretinoin on tissue strength and skin components in a murine incisional wound model. AB - BACKGROUND: Vitamin A derivatives modulate the inflammatory phase in wound healing. Retinoic acid can restore decreased tensile strength and collagen content in steroid- or diabetes-impaired wounds. It is hypothesized that retinoic acid can lead to accelerated healing with improved breaking strength in unimpaired incisional wounds. MATERIALS AND METHODS: Skin incisions were made in 45 CD-1 mice. The sutured wounds were treated once daily with topical all-trans retinoic acid 0.1% (n = 15), vehicle ointment (n = 15), or left untreated (n = 15). Skin biopsies at 1-, 2-, and 3-week intervals were examined using hematoxylin and eosin (H&E), Masson's trichrome, and immunoperoxidase staining methods. Wound breaking strength was determined by biomechanical analysis. RESULTS: Incisions treated with retinoic acid exhibited a significantly reduced breaking strength at week 1 when compared to the vehicle and control group. Histologic examination showed a prolonged inflammatory reaction with abundant deposition of granulation tissue. Despite an increased fibroplastic proliferation in the tretinoin-treated wounds, the production of collagen was diminished. CONCLUSIONS: Topical retinoic acid does not enhance the healing of unimpaired incisional wounds. The inadequate tensile strength in the early phase of the healing process is possibly the result of an increased dermal inflammatory response and the decreased collagen content. Although these adverse effects disappeared by 3 weeks postwounding, we found no discernible benefit of supplemental retinoic acid in unimpaired wounds. PMID- 15793507 TI - Use of complementary medicine among outpatients with dermatologic conditions within Yorkshire and South Wales, United Kingdom. AB - BACKGROUND: The use of complementary and alternative medicine (CAM) is increasing throughout the Western world, particularly by patients with chronic disease. In 1999, 20% of the United Kingdom population reported that they had used CAM in the preceding 12 months. OBJECTIVES: We sought to investigate the use of CAM by outpatients with general dermatologic conditions in 3 United Kingdom cities: Leeds (North England), and Cardiff and Swansea (South Wales). METHODS: Independently constructed, anonymous, self-directed questionnaires were designed and distributed to 400 consecutive outpatients with dermatologic conditions in Leeds and 637 consecutive outpatients with dermatologic conditions in Cardiff and Swansea. RESULTS: In Leeds 302/400 questionnaires were fully completed with 39% patients having used CAM. In South Wales 415/637 questionnaires were fully completed with 34% having used CAM. Of these, 45% of patients in Leeds and 50% of patients in South Wales were using CAM to treat their dermatologic condition. The most popular treatments in Leeds, Cardiff, and Swansea were herbal medicine and homeopathy. LIMITATIONS: These results were obtained from outpatients with dermatologic conditions in the United Kingdom and may not be applicable to other populations. CONCLUSIONS: More than one third of outpatients with general dermatologic conditions in two geographically distant regions are using CAM. More than 45% of these patients used CAM to treat their dermatologic condition. PMID- 15793508 TI - Persistent erythema and edema of the midthird and upper aspect of the face (morbus morbihan): evidence of hidden immunologic contact urticaria and impaired lymphatic drainage. AB - BACKGROUND: A persistent erythema and edema of the midthird and upper aspect of the face, which bears some resemblance to Melkersson-Rosenthal syndrome and rosaceous lymphedema, has been characterized as morbus morbihan (MM) by French dermatologists. The disease of yet unknown cause starts with recurrent facial edema of short duration, which ultimately leads to persistent swelling after a period of weeks or months. METHODS: We recruited 6 patients with MM and acquired their history, routine blood tests, and individual UV light tolerability. To check for contact allergies the allergen patch test and the open epicutaneous patch test were performed. To objectify the skin conditions laser Doppler flowmetry and 20-MHz ultrasound were used. Five patients with similar symptoms, but with definitely transient facial erythema and edema caused by proven contact urticaria on cosmetics served as a comparison group. RESULTS: In all patients, routine blood tests and UVA/UVB light tests showed no pathologic results. Observations of 6 patients with MM revealed the common feature of a clinically relevant immunologic contact urticaria caused by various cosmetic ingredients, which could be diagnosed in all of them. Delayed resorption of the acute edema and prolonged inflammation were shown by laser Doppler flowmetry and 20-MHz ultrasound in the affected skin areas in patients with MM after induction of immunologic contact urticaria by a cosmetic ingredient. Strict avoidance of cosmetics yielded a remarkable clinical benefit in the follow-up examinations. CONCLUSIONS: We conclude that recurrent and possibly subclinical inflammation caused by immunologic contact urticaria in conjunction with a locally pre existing lowered lymphatic drainage plays a crucial role in the evolution of MM. PMID- 15793509 TI - Clinical spectrum and virologic characteristics of anal intraepithelial neoplasia in HIV infection. AB - BACKGROUND: Anal intraepithelial neoplasia (AIN) represents a precursor lesion of invasive squamous cell carcinoma with a clear association to high-risk human papillomavirus (HPV) types. HIV infection is strongly associated with a higher prevalence of genital HPV infection, a higher incidence of AIN, and, consecutively, an increased risk for anal cancer. OBJECTIVE: The aim of this study was to determine the clinical spectrum of AIN and lesional HPV colonization in a cohort of homosexual men who were HIV positive and had a history of receptive anal intercourse. METHODS: In all, 103 men who were HIV-1 positive were screened by using clinical, proctologic, cytologic, histologic, and HPV DNA testing. RESULTS: Of all patients, 86% had anal HPV infection at their first visit. HPV-16 (53%), HPV-18 (27%), HPV-58 (22%), and HPV-83 (22%) were the most frequently found HPV types. AIN was diagnosed in 20 of the 103 patients (19.4%). High-risk HPV types were present in all AIN cases with up to 7 different high risk and up to 5 different low-risk types per lesion. Histologically, 7 (35%), 7 (35%), and 6 (30%) of the patients had AIN grade I, II, or III, respectively. Four different types of clinical presentation could be distinguished in the 20 patients with AIN: bowenoid (1 case, 5%); erythroplakic (2 cases, 10%); verrucous (6 cases, 30%); and leukoplakic (11 cases, 55%). All verrucous lesions were graded as high-grade intraepithelial lesions in cytology, whereas 6 of the 11 leukoplakic lesions (55%) were low grade. All verrucous AIN carried at least 4 different HPV types, always including HPV-16, and the mean number of HPV types was higher in verrucous lesions than in leukoplakic lesions (5.5 vs 3.8, respectively). CONCLUSION: These data confirm the high incidence and prevalence of AIN in patients who are HPV positive with HIV infection. Four different clinical types of AIN can be distinguished that might have prognostic implications. Standardized screening programs for anal cancer prevention and treatment protocols for AIN in patients infected with HIV must be implemented. PMID- 15793510 TI - Assessing the role of race in quantitative measures of skin pigmentation and clinical assessments of photosensitivity. AB - BACKGROUND: Given the increasing demographic diversity in the United States, clarifying relationships between race, color, ethnicity, and disease processes is critical. OBJECTIVES: We sought to examine the correlation between objective measures of skin pigmentation, racial identification, and physician-diagnosed and self-reported skin phototypes. METHODS: A total of 558 participants (76 nonwhite) were evaluated. A subset underwent spectrometric readings and digital photography of the upper aspect of the inner arm. Self-identified race was compared with 7 measures of pigmentation. RESULTS: Race correlates best with physician-diagnosed skin phototype (r = 0.55, P < .01), whereas self-reported skin phototype, spectrometry, and colorimetry correlate poorly with race (r = 0.28, < 0.40, and r > -0.31, respectively, P < .01). Associations between race and subjective measures strengthen among patients with darker skin. CONCLUSION: Objective measures of pigmentation fail to correlate well with race, whereas race correlates moderately with physician-diagnosed skin phototype. Including objective methods of analyzing skin color may reduce subjective influences of race in assessing photosensitivity and potential risk for skin cancer. PMID- 15793511 TI - Kaposiform hemangioendothelioma without Kasabach-Merritt phenomenon. AB - Kasabach-Merritt phenomenon is a serious coagulopathy associated with kaposiform hemangioendothelioma (KHE), tufted angioma, and possibly other vascular neoplasms. KHE presenting in the absence of Kasabach-Merritt phenomenon is rare, although tufted angioma frequently occurs without thrombocytopenia. We retrospectively reviewed 10 cases of KHE without Kasabach-Merritt phenomenon. The tumors appeared as soft tissue masses with the overlying skin being either normal, erythematous, or violaceous. There were no radiologic or microscopic differences in noncoagulopathic KHE as compared with coagulopathic KHE. Evidence of platelet trapping and hemosiderin deposition was seen histologically, despite normal serum platelet levels. All KHE were less than 8 cm in diameter, suggesting that tumors that grow no larger than this size are less likely to trap platelets in sufficient quantity to cause thrombocytopenia. Our series confirms that KHE appears with a wide spectrum of behavior and response to treatment. The decision as to whether or not to treat a noncoagulopathic KHE should be based on the size and location of the tumor and the possible side effects of therapy. PMID- 15793512 TI - Vaccination-induced cutaneous pseudolymphoma. AB - BACKGROUND: Although mild early cutaneous transient reactions to vaccinations are common, late-onset chronic lesions have been scarcely reported. We report herein a series of 9 patients presenting with cutaneous and subcutaneous pseudolymphoma. OBSERVATIONS: Nine patients presenting with late-onset, chronic skin lesions occurring at the site of antihepatitis B (8 cases) and antihepatitis A (one case) vaccination were reported. Histopathologic and immunohistochemic studies, and molecular analysis of clonality of skin biopsy specimens, were performed. Furthermore, the presence of vaccine products was investigated in skin lesions by using histochemical, microanalytic, and electronic microscopy techniques. RESULTS: Histopathologic studies showed dermal and hypodermal lymphocytic follicular infiltrates with germinal center formation. The center of follicles was mostly composed of B cells without atypia, whereas CD4+ T cells were predominant at the periphery. Molecular analysis of clonality revealed a polyclonal pattern of B-cell and T-cell subsets. Aluminium deposits were evidenced in all cases by using histochemical staining in all cases, and by microanalysis and ultrastructural studies in one case. Associated manifestations were vitiligo (one case) and chronic fatigue with myalgia (two cases). CONCLUSION: Cutaneous lymphoid hyperplasia is a potential adverse effect of vaccinations including aluminium hydroxide as an adjuvant. Further prospective studies are warranted to evaluate the incidence of this complication in the immunized population. PMID- 15793513 TI - Surgical repair of temple defects after Mohs micrographic surgery. AB - BACKGROUND: It is typically recommended that linear surgical closures follow a relaxed skin tension line (RSTL). In the temple, these lines generally run parallel to the orbital rim. However, closures parallel to RSTLs are not feasible for many medium and large surgical defects because of anatomic constraints. OBJECTIVES: We sought to describe our method of repairing temple defects and assess the cosmetic outcome of temple defects repaired perpendicular to the RSTLs. METHODS: We performed a two-phase study of temple closures. In the first phase, a retrospective analysis was performed to assess the defect size and type of closure used for 99 consecutive temple defects. In the second phase, 27 of 86 patients who previously underwent linear surgical repairs in the temple that were designed perpendicular to the RSTLs were evaluated 12 to 24 months postoperatively to assess the cosmetic outcome. Results In the first phase of the study, there was an inverse correlation between the width of the postoperative defect and the ability to close the defect parallel to the RSTLs. The percentage of defects that could be repaired parallel to the RSTLs for defects up to 1 cm in width, 1.1 to 2 cm in width, and greater than 2 cm in width was 46%, 28%, and 8%, respectively. In the second phase of the study, 27 patients with temple defects repaired perpendicular to the RSTLs were evaluated an average of 1.7 years after operation. Most scars (83%) were clinically invisible or barely visible. There were no cases with residual standing cones (dog-ears) or hypertrophic scars. CONCLUSION: The vast majority of temple defects can be repaired in a linear fashion. Smaller defects (width < 1 cm) can be repaired parallel to the RSTL. Those that are larger (width 1-3 cm) or more medially located can be repaired perpendicular to the RSTL, using loose cheek skin for the closure. Very large defects (width > 3-5 cm) or those with limited cheek mobility may require flaps or skin grafts for closure. PMID- 15793514 TI - Primary cicatricial alopecia: histopathologic findings do not distinguish clinical variants. AB - BACKGROUND: Primary cicatricial alopecias encompass a group of disorders characterized by permanent destruction of the hair follicle. The varied clinical features and differences in terminology have led to difficulties in defining consistent clinicopathologic correlation. OBJECTIVE: We sought clinicopathologic correlation of 6 clinically distinct primary cicatricial alopecias: lichen planopilaris, frontal fibrosing alopecia, pseudopelade (Brocq), central centrifugal alopecia, folliculitis decalvans, and tufted folliculitis. METHODS: We conducted prospective and blinded histopathologic evaluation of clinically typical primary cicatricial alopecias. Biopsy specimens were taken from early affected scalp lesions and paired with samples from clinically unaffected areas in the same patient. RESULTS: The lymphocytic and neutrophilic groups were readily distinguished histologically. However, within the two groups clinically distinct primary cicatricial alopecias could not be distinguished with current histopathologic techniques. CONCLUSION: A descriptive, standardized histopathologic reporting of follicular architecture, type, location, and extent of the inflammatory infiltrate, and presence or absence of sebaceous glands, may be of greatest value in guiding the treatment of patients with primary cicatricial alopecias. PMID- 15793515 TI - Ossifying fibromyxoid tumor of the skin: a report of 2 cases with light microscopic, immunohistochemical, and electron microscopic characterization. AB - Ossifying fibromyxoid tumor is a rare fibro-osseous neoplasm occurring predominantly in the subcutaneous tissue and/or skeletal muscle of the extremities. The immunohistochemical and ultrastructural studies in reported cases favor neural or Schwannian origin. To date, ossifying fibromyxoid tumor presenting as a primary skin tumor has not been described in the literature. Two cases of primary cutaneous ossifying fibromyxoid tumor with light microscopic, immunohistochemical, and ultrastructural characterization are presented. Our observations also support neural differentiation of these tumors. PMID- 15793516 TI - Microscopic leprosy skin lesions in primary neuritic leprosy. AB - The histologic diagnosis of primary neuritic leprosy (PNL) remains a public health care concern, especially when nerve biopsies cannot be performed. As such, some authors emphasize the importance of performing a skin biopsy of a hypoesthetic area even without clinically visible lesions. In this study, an attempt was made to define the histologic changes in the sensory altered skin of 42 clinically diagnosed PNL patients. Histologic alterations caused by leprosy were seen in 31% of these patients: 6 were classified as borderline tuberculoid and 7 as indeterminate. In addition, 33% showed mild, non-specific, mononuclear cell infiltrates around the blood vessels within the papillary and reticular dermis that probably reflected an early inflammatory reaction to Mycobacterium leprae infection. Only 36% of those biopsied had no significant lesions. Our results suggested that, while not all PNL patients are similar, histologic skin examination can contribute to early leprosy detection and commencement of adequate treatment. PMID- 15793517 TI - Cutaneous alternariosis in transplant recipients: clinicopathologic review of 9 cases. AB - OBJECTIVES: We sought to evaluate and review the clinical and histopathologic features of cutaneous infections caused by the environmental opportunistic fungus Alternaria observed in transplant recipients. METHODS: We conducted a retrospective study of cases of cutaneous alternariosis in transplant recipients given a diagnosis in 3 hospitals in Catalonia, Spain, between 1991 and 2001. The clinical and evolution features were reviewed. A panel of histopathologic features was evaluated by two independent observers in all cutaneous biopsy specimens. RESULTS: In all, 9 transplant recipients (8 men and 1 woman) presenting opportunistic cutaneous alternariosis were studied. The patients were 4 renal, 2 cardiac, 1 liver, and 2 lung transplant recipients. All patients were treated with different immunosuppressive therapeutic regimes. The lesions were solitary (3 patients) or multiple grouped (6 patients): papules (4 patients), plaques (5 patients), inflammatory nodules (2 patients), and recurrent cellulitis with secondary ulceration (1 patient), mainly located on the lower extremities. No extracutaneous involvement was detected. A previous traumatic event was recorded in two patients. A total of 12 cutaneous biopsy specimens were reviewed. Biopsy specimens from early lesions (<3 months evolution) were often characterized by the presence of epidermal changes (3/6 pseudoepitheliomatous hyperplasia; 50%), a diffuse dermal mixed inflammatory infiltrate of lymphocytes, plasma cells, histiocytes, neutrophils, and giant cells, and rare and focal granuloma formation. Dermal abscess or necrotizing folliculitis was occasionally noted. In biopsy specimens from more advanced lesions (>3 months evolution), the presence of a granulomatous inflammatory infiltrate was a constant feature. Suppurative granulomas (2/6; 33%) and sarcoidlike granulomas (2/6; 33%) were noted. In all biopsy specimens, fungal structures with a typical round-to-oval, thick refractile wall were identified. CONCLUSION: Different clinical and histopathologic patterns can be noted in cutaneous alternariosis. Clinically the lesions manifest as solitary or grouped papules, plaques, or nodules mainly involving the lower extremities. Histologically, a relationship between the evolution of the cutaneous lesions and granuloma formation is detected. An increased awareness regarding the clinical and histopathologic features of cutaneous alternariosis in transplant recipients is important to achieve early detection and treatment. PMID- 15793519 TI - Biologic treatments for psoriasis. PMID- 15793518 TI - Narrowband UVB phototherapy in skin conditions beyond psoriasis. AB - BACKGROUND: Narrowband (NB) UVB phototherapy has been proven to be clearly more effective than broadband UVB and safer and/or more practicable than psoralen-UVA in the management of psoriasis. However, the role of NB UVB seems to be less clear in the management of skin conditions beyond psoriasis. OBJECTIVES: We sought to give an update on clinical experiences in NB UVB of nonpsoriatic skin conditions, and to establish its current position within the spectrum of competing photo(chemo)therapeutic options. METHODS: The computerized bibliographic database PubMed, without time limits, and other sources were screened for clinical trials on NB UVB. Included were research articles of randomized controlled trials, open prospective studies, and retrospective observations on NB UVB in skin disorders other than psoriasis. RESULTS: A total of 28 articles met our eligibility criteria including 6 randomized controlled studies, 16 open prospective studies, and 6 retrospective observations. NB UVB is effective in patients with chronic atopic dermatitis (AD) (n = 719) and generalized vitiligo (n = 305) and appears to have some advantages over competing photo(chemo)therapeutic regimens. NB UVB also seems to be effective in patients with polymorphic light eruption (n = 25), early stages of cutaneous T-cell lymphoma (n = 108), chronic urticaria (n = 88), lichen planus (n = 15), pruritus associated with polycythemia vera (n = 10), seborrheic dermatitis (n = 18), actinic prurigo (n = 6), and acquired perforating dermatosis (n = 5). The quality of evidence determined for the aforementioned diagnoses ranged from high to moderate to very low. CONCLUSIONS: The best currently available data on NB UVB in nonpsoriatic conditions exist for AD and generalized vitiligo. In view of its efficacy, benefit/risk profile, and costs, NB UVB may be considered the first line photo(chemo)therapeutic option for moderately severe AD and widespread vitiligo. In the treatment of most other nonpsoriatic conditions, NB UVB appears to be effective, but current data allow no definitive conclusions as to whether NB UVB should be preferred to competing photo(chemo)therapeutic options such as UVA1 and psoralen-UVA regimens. Because NB UVB may have a wider indication spectrum, including AD, vitiligo, and early-stage T-cell lymphoma, and appears to be equally effective or even more effective than broadband UVB, a switch from broadband UVB to NB UVB seems to be justified. PMID- 15793520 TI - Panniculitis. PMID- 15793521 TI - Surgical pearl: safe splinting positions for skin grafts on the hand and wrist. PMID- 15793522 TI - Medical pearl: errors in itch. PMID- 15793523 TI - Surgical pearl: dermabrasion for the correction of incomplete cleft earlobe. PMID- 15793524 TI - Surgical pearl: the insulated Hyfrecator needle: a no-cost surgical safety tip. PMID- 15793525 TI - Treatment of atopic dermatitis with herbal combination of Eleutherococcus, Achillea millefolium, and Lamium album has no advantage over placebo: a double blind, placebo-controlled, randomized trial. AB - OBJECTIVE: To test the efficacy of tri-herbal combination on atopic dermatitis in a randomized, placebo-controlled trial. METHODS: Forty-nine patients were included for 2 weeks of treatment. Patients were followed until week 8. RESULTS: The response to the real medication was significant in objective and subjective parameters. Patients maintained partial remission until the end of follow-up. The placebo-treated group had a similar response without a significant difference. CONCLUSIONS: Treatment with tri-herbal combination for atopic dermatitis does not differ from treatment with placebo. PMID- 15793526 TI - Papular mycosis fungoides: a new clinical variant of early mycosis fungoides. AB - BACKGROUND: Mycosis fungoides (MF) is the most common type of cutaneous T-cell lymphoma. In early stages of the disease many different clinicopathologic variants have been observed. OBSERVATION: We report 6 patients with early manifestations of MF characterized by the sole presence of papules which, unlike the papules of lymphomatoid papulosis, did not show a tendency for spontaneous resolution. Histologic examination confirmed the diagnosis of MF in all cases. Immunohistochemical staining for CD30 was negative in all cases. Follow-up data showed the nonaggressive behavior of the disease, confirming that the lesions were not manifestations of advanced MF. CONCLUSION: Papular MF is a new variant of early MF characterized by the presence of papules in the absence of more conventional early lesions (patches) of the disease. This variant should be added to the long list of clinicopathologic subtypes of MF. PMID- 15793527 TI - No long-term changes in psoriasis severity and quality of life following climate therapy. AB - This paper describes disease severity and quality of life of 286 patients who underwent climate therapy. The lowest disease severity and better life quality occurred 2 weeks after therapy. However, by 4 and 8 months after therapy, all measures had returned to approximately the same level as before treatment. PMID- 15793528 TI - Leprosy masquerading as lupus. PMID- 15793529 TI - Simultaneous chronic cutaneous infection with Mycobacterium marinum and Nocardia asteroides. PMID- 15793530 TI - Soft tissue infection caused by Gemella morbillorum in two intravenous drug users. PMID- 15793531 TI - Mycoplasma pneumoniae infection associated with bullous erythema multiforme. PMID- 15793532 TI - A case of poikiloderma vasculare atrophicans, a rare variant of cutaneous T-cell lymphoma, responding to extracorporeal photopheresis. PMID- 15793533 TI - T/natural killer-cell lymphomas. PMID- 15793534 TI - Bone marrow clonal T-cell population in lymphomatoid papulosis. PMID- 15793535 TI - Acrokeratosis paraneoplastica (Bazex syndrome) preceeding the diagnosis of metastatic squamous cell carcinoma of the esophagus. PMID- 15793536 TI - Interstitial granulomatous drug reaction caused by Chinese herbal medication. PMID- 15793537 TI - Long-term complete remission of cutaneous melanoma metastases in association with a folk remedy. PMID- 15793538 TI - Vitiligo-like hypopigmentation associated with imiquimod treatment of genital warts. PMID- 15793539 TI - Infiltrative squamous cell carcinoma on the scalp after treatment with 5% imiquimod cream. PMID- 15793540 TI - A case of mucocutaneous Behcet's disease responding to etanercept. PMID- 15793541 TI - Crusted scabies in a patient being treated with infliximab for juvenile rheumatoid arthritis. PMID- 15793542 TI - Tacrolimus ointment in the treatment of dirty neck lesions in atopic dermatitis. PMID- 15793543 TI - Treatment of pyostomatitis vegetans with topical tacrolimus. PMID- 15793544 TI - Three cases of Bowen's disease on the lower abdomen associated with high-risk types 16, 33, and 59 of human papillomavirus. PMID- 15793545 TI - Nevus sebaceus with psammomatous calcified spherules in the apocrine glands. PMID- 15793546 TI - Comel-Netherton syndrome in brothers and expression of cytokeratins. PMID- 15793547 TI - Painful subungal dyskeratotic tumors in incontinentia pigmenti. PMID- 15793548 TI - Necrobiotic xanthogranuloma: isolated skeletal muscle involvement and unusual changes. PMID- 15793549 TI - Multiple glomangiomas: potential for confusion with blue rubber bleb nevus syndrome. PMID- 15793550 TI - Acute generalized exanthematous pustulosis caused by piperacillin/tazobactam. PMID- 15793551 TI - What determines whether patients with skin conditions visit dermatologists? PMID- 15793552 TI - False firstedness: the dilemma of priority claims in biomedical publications. PMID- 15793553 TI - A dictionary of dermatology should serve to clarify terminology but not be yet another textbook of skin diseases! PMID- 15793554 TI - Remission and time of resolution of nail psoriasis during infliximab therapy. PMID- 15793555 TI - Randomized controlled trial of pimecrolimus 1% cream for treatment of facial acne vulgaris. PMID- 15793556 TI - Bioremediation of crude oil-polluted soil--effect of poultry droppings and natural rubber processing sludge application on biodegradation of petroleum hydrocarbons. AB - Laboratory bioremediation experiments were carried out on crude oil-polluted soil samples by applying various amounts of poultry droppings and natural rubber processing sludge as nutrient supplements at 29 degrees and using slurry-phase and solid-phase biodegradation techniques. Changes in the total hydrocarbon content of the soil were determined using a spectrophotometric technique as a function of time. It was found that the extent of crude oil degradation in untreated soil samples was markedly lower (by up to 100%) than in the soil samples treated with nutrient supplements. Hydrocarbon degradation efficiency was higher in the slurry-phase than in the soil-phase technique. PMID- 15793557 TI - Effects of organophosphorous pesticides used in china on various mammalian cells. AB - Organophosphorous pesticides are currently widely used in China to help boost agricultural production. However, these pesticides pose various threats to organisms, including humans, and are thus a cause of concern. Five organophosphorous pesticides, monocrotophos, omethoate, parathion-methyl, phoxim and dichlorvos, were examined for their effects on mammalian cell lines to determine their potential impact on physiological functions in vivo. Results show an increased proliferation of MCF-7 cells treated with 0.2 microM monocrotophos or 0.4 microM omethoate, suggesting that these compounds can induce breast cancer cell proliferation at relatively low concentrations. Murine primary spleen cells markedly decreased in number starting at a pesticide concentration of 0.01 microM; no cytotoxicity was observed below 0.001 microM. BALB/c3T3 murine fibroblasts treated with 0.25 microM monocrotophos showed enhanced DNA synthesis, while those treated with the other pesticides showed results similar to that of the control. The different pesticides reduced the acetylcholinesterase (AChE) activity of the rat neuronal cell line PC12 in a dose-dependent manner up to 100 microM. Parathion-methyl and phoxim showed acute toxicity at 0.01 microM. Finally, phoxim and parathion-methyl significantly reduced the transepithelial electrical resistance (TEER) of human intestinal Caco-2 cells, indicating that these pesticides can disrupt the tight-junction permeability of cell monolayers. These in vitro assays, which are rapid, reproducible, simple and inexpensive, clearly show the effects of organophosphorous pesticides on mammalian cells and suggest the potential impact of these pesticides on organisms in vivo. PMID- 15793558 TI - Simulation of the toxicokinetics of trichloroethylene, methylene chloride, styrene and n-hexane by a toxicokinetics/toxicodynamics model using experimental data. AB - The toxicokinetics/toxicodynamics (TKTD) model simulates the toxicokinetics of a chemical based on physiological data such as blood flow, tissue partition coefficients and metabolism. In this study, Andersen and Clewell's TKTD model was used with seven compartments and ten differential equations for calculating chemical balances in the compartments (Andersen and Clewell 1996, Workshop on physiologically-based pharmacokinetic/pharmacodynamic modeling and risk assessment, Aug. 5-16 at Colorado State University, U.S.A) . Using this model, the authors attempted to simulate the behavior of four chemicals: trichloroethylene, methylene chloride, styrene and n-hexane, and the results were evaluated. Simulations of the behavior of trichloroethylene taken in via inhalation and oral exposure routes were also done. The differences between simulations and measurements are due to the differences between the absorption rates of the exposure routes. By changing the absorption rates, the simulation showed agreement with the measured values. The simulations of the other three chemicals showed good results. Thus, this model is useful for simulating the behavior of chemicals for preliminary toxicity assessment. PMID- 15793559 TI - Effect of soil acidification on the growth of Korean pine (Pinus koraiensis) seedlings in a granite-derived forest soil. AB - The growth of pine trees has diminished in recent years in industrial areas of Korea. Soil acidification is believed to be responsible. To study its effects, we grew seedlings of three-year-old Korean pine in brown forest soil derived from granite, which had been treated with an acid solution, for 182 days. The anion mol ratio in the solution was SO4(2-):NO3-:Cl-=5:3:2, which is the average in the total precipitation in Korea; six H+ ion concentrations in the soil were studied (0 (control), 10, 30, 60 and 90 mmol H+.kg-1). With increasing amounts of H+ added to the soil, the concentrations of Ca, Mg, K, Al and Mn increased, especially below a soil pH of 3.8. The concentrations of Ca, Mg and K in pine needles and stems increased with increasing H+ added to the soil, whereas their concentrations in the root decreased. Conversely, the concentration of N and P in each organ of the pine plant was higher in all treatments than in controls. Also, the concentrations of Al and Mn increased significantly in all organs of the plant with increasing H+. We also estimated the effect of deliberate soil acidification on tree growth, using the molar ratio (Ca+Mg+K)/Al as an indicator of soil acidification. A strong positive correlation was found between the total dry mass (TDM) of seedlings and the (Ca+Mg+K)/Al molar ratio calculated from the concentrations of water-soluble elements in soil (r=0.99, p<0.001). When the (Ca+Mg+K)/Al molar ratio reached 1.0, the relative TDM had fallen to 40%. These results show that deliberate soil acidification reduces the growth of the Korean pine less than it does the Red pine, which has been the dominant species in Korea. PMID- 15793560 TI - Methods of analysis for chemicals that disrupt cellular signaling pathways: risk assessment for potential endocrine disruptors. AB - Here we present a basic concept and several examples of methods of analysis for chemicals that disrupt cellular signaling pathways, in view of risk assessment for potential endocrine disrupting chemicals (EDCs). The key cellular signaling pathways include 1) ER/coactivator interaction, 2) AR translocation into the nucleus, 3) ER/NO/sGC/cGMP, 4) ER/Akt, 5) ER/Src, 6)ER/Src/Grb2, and 7) ER/Ca2+/CaM/CaMK pathways. These were visualized in relevant live cells using newly developed fluorescent and bioluminescent probes. Changes in cellular signals were thereby observed in nongenomic pathways of steroid hormones upon treatment of the target cells with steroid hormones and related chemicals. This method of analysis appears to be a rational approach to high-throughput prescreening (HTPS) of biohazardous chemicals, EDCs, in particular. Also described was the screening of gene expression by serial analysis of gene expression and gene chips upon applying EDCs to breast cancer cells, mouse livers, and human neuroblastoma NB-1 cells. PMID- 15793561 TI - A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. AB - Myeloproliferative disorders are clonal haematopoietic stem cell malignancies characterized by independency or hypersensitivity of haematopoietic progenitors to numerous cytokines. The molecular basis of most myeloproliferative disorders is unknown. On the basis of the model of chronic myeloid leukaemia, it is expected that a constitutive tyrosine kinase activity could be at the origin of these diseases. Polycythaemia vera is an acquired myeloproliferative disorder, characterized by the presence of polycythaemia diversely associated with thrombocytosis, leukocytosis and splenomegaly. Polycythaemia vera progenitors are hypersensitive to erythropoietin and other cytokines. Here, we describe a clonal and recurrent mutation in the JH2 pseudo-kinase domain of the Janus kinase 2 (JAK2) gene in most (> 80%) polycythaemia vera patients. The mutation, a valine to-phenylalanine substitution at amino acid position 617, leads to constitutive tyrosine phosphorylation activity that promotes cytokine hypersensitivity and induces erythrocytosis in a mouse model. As this mutation is also found in other myeloproliferative disorders, this unique mutation will permit a new molecular classification of these disorders and novel therapeutical approaches. PMID- 15793562 TI - Peak SIV replication in resting memory CD4+ T cells depletes gut lamina propria CD4+ T cells. AB - In early simian immunodeficiency virus (SIV) and human immunodeficiency virus-1 (HIV-1) infections, gut-associated lymphatic tissue (GALT), the largest component of the lymphoid organ system, is a principal site of both virus production and depletion of primarily lamina propria memory CD4+ T cells; that is, CD4 expressing T cells that previously encountered antigens and microbes and homed to the lamina propria of GALT. Here, we show that peak virus production in gut tissues of SIV-infected rhesus macaques coincides with peak numbers of infected memory CD4+ T cells. Surprisingly, most of the initially infected memory cells were not, as expected, activated but were instead immunophenotypically 'resting' cells that, unlike truly resting cells, but like the first cells mainly infected at other mucosal sites and peripheral lymph nodes, are capable of supporting virus production. In addition to inducing immune activation and thereby providing activated CD4+ T-cell targets to sustain infection, virus production also triggered an immunopathologically limiting Fas-Fas-ligand-mediated apoptotic pathway in lamina propria CD4+ T cells, resulting in their preferential ablation. Thus, SIV exploits a large, resident population of resting memory CD4+ T cells in GALT to produce peak levels of virus that directly (through lytic infection) and indirectly (through apoptosis of infected and uninfected cells) deplete CD4+ T cells in the effector arm of GALT. The scale of this CD4+ T-cell depletion has adverse effects on the immune system of the host, underscoring the importance of developing countermeasures to SIV that are effective before infection of GALT. PMID- 15793563 TI - Massive infection and loss of memory CD4+ T cells in multiple tissues during acute SIV infection. AB - It has recently been established that both acute human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) infections are accompanied by a dramatic and selective loss of memory CD4+ T cells predominantly from the mucosal surfaces. The mechanism underlying this depletion of memory CD4+ T cells (that is, T-helper cells specific to previously encountered pathogens) has not been defined. Using highly sensitive, quantitative polymerase chain reaction together with precise sorting of different subsets of CD4+ T cells in various tissues, we show that this loss is explained by a massive infection of memory CD4+ T cells by the virus. Specifically, 30-60% of CD4+ memory T cells throughout the body are infected by SIV at the peak of infection, and most of these infected cells disappear within four days. Furthermore, our data demonstrate that the depletion of memory CD4+ T cells occurs to a similar extent in all tissues. As a consequence, over one-half of all memory CD4+ T cells in SIV-infected macaques are destroyed directly by viral infection during the acute phase-an insult that certainly heralds subsequent immunodeficiency. Our findings point to the importance of reducing the cell-associated viral load during acute infection through therapeutic or vaccination strategies. PMID- 15793564 TI - Golgi-localized GAP for Cdc42 functions downstream of ARF1 to control Arp2/3 complex and F-actin dynamics. AB - The small GTP-binding ADP-ribosylation factor 1 (ARF1) acts as a master regulator of Golgi structure and function through the recruitment and activation of various downstream effectors. It has been proposed that members of the Rho family of small GTPases also control Golgi function in coordination with ARF1, possibly through the regulation of Arp2/3 complex and actin polymerization on Golgi membranes. Here, we identify ARHGAP10--a novel Rho GTPase-activating protein (Rho GAP) that is recruited to Golgi membranes through binding to GTP-ARF1. We show that ARHGAP10 functions preferentially as a GAP for Cdc42 and regulates the Arp2/3 complex and F-actin dynamics at the Golgi through the control of Cdc42 activity. Our results establish a role for ARHGAP10 in Golgi structure and function at the crossroads between ARF1 and Cdc42 signalling pathways. PMID- 15793565 TI - Stat3-induced apoptosis requires a molecular switch in PI(3)K subunit composition. AB - Physiological apoptosis is induced by a switch from survival to death signalling. Dysregulation of this process is frequently associated with cancer. A powerful model for this apoptotic switch is mammary gland involution, during which redundant milk-producing epithelial cells undergo apoptosis. Signal transducer and activator of transcription 3 (Stat3) is an essential mediator of this switch but the mechanism has not yet been defined. Stat3-dependent cell death during involution can be blocked by activation of Akt/protein kinase B (PKB), a downstream effector of the phosphoinositide-3-OH kinase (PI(3)K) pathway. Here we show that expression of the PI(3)K regulatory subunits p55alpha and p50alpha is induced by Stat3 during involution. In the absence of Stat3 in vivo, upregulation of p55alpha and p50alpha is abrogated, levels of activated Akt are sustained and apoptosis is prevented. Chromatin immunoprecipitation assays show that Stat3 binds directly to the p55alpha and p50alpha promoters in vivo. Overexpression of either p55alpha or p50alpha reduces levels of activated Akt. We propose a novel mechanism in which Stat3 regulates apoptosis by inducing expression of distinct PI(3)K regulatory subunits to downregulate PI(3)K-Akt-mediated survival signalling. PMID- 15793566 TI - CSN facilitates Cullin-RING ubiquitin ligase function by counteracting autocatalytic adapter instability. AB - The COP9 signalosome (CSN) is known to bind cullin-RING ubiquitin ligases (CRLs) and to promote their activity in vivo. The mechanism of this stimulation has remained enigmatic because CSN's intrinsic and associated enzymatic activities paradoxically inhibit CRL activity in vitro. Reconciling this paradox, we show here that Csn5-catalysed cullin (Cul) deneddylation and Ubp12-mediated deubiquitination cooperate in maintaining the stability of labile substrate adapters, thus facilitating CRL function. Various fission-yeast csn and ubp12 deletion mutants have lower levels of the Cul3p adapter Btb3p. This decrease is due to increased autocatalytic, Cul3p-dependent, ubiquitination and the subsequent degradation of Btb3p. The CSN-Ubp12p pathway also maintains the stability of the Cul1p adapter Pop1p, a mechanism required for the efficient destruction of its cognate substrate Rum1p. Emphasizing the physiological importance of this mechanism, we found that the dispensable csn5 and ubp12 genes become essential for viability when adapter recruitment to Cul1p is compromised. Our data suggest that maintenance of adapter stability is a general mechanism of CRL control by the CSN. PMID- 15793567 TI - SMC5 and SMC6 genes are required for the segregation of repetitive chromosome regions. AB - Structure chromosome (SMC) proteins organize the core of cohesin, condensin and Smc5-Smc6 complexes. The Smc5-Smc6 complex is required for DNA repair, as well as having another essential but enigmatic function. Here, we generated conditional mutants of SMC5 and SMC6 in budding yeast, in which the essential function was affected. We show that mutant smc5-6 and smc6-9 cells undergo an aberrant mitosis in which chromosome segregation of repetitive regions is impaired; this leads to DNA damage and RAD9-dependent activation of the Rad53 protein kinase. Consistent with a requirement for the segregation of repetitive regions, Smc5 and Smc6 proteins are enriched at ribosomal DNA (rDNA) and at some telomeres. We show that, following Smc5-Smc6 inactivation, metaphase-arrested cells show increased levels of X-shaped DNA (Holliday junctions) at the rDNA locus. Furthermore, deletion of RAD52 partially suppresses the temperature sensitivity of smc5-6 and smc6-9 mutants. We also present evidence showing that the rDNA segregation defects of smc5/smc6 mutants are mechanistically different from those previously observed for condensin mutants. These results point towards a role for the Smc5 Smc6 complex in preventing the formation of sister chromatid junctions, thereby ensuring the correct partitioning of chromosomes during anaphase. PMID- 15793568 TI - Spinophilin regulates Ca2+ signalling by binding the N-terminal domain of RGS2 and the third intracellular loop of G-protein-coupled receptors. AB - Signalling by G proteins is controlled by the regulator of G-protein signalling (RGS) proteins that accelerate the GTPase activity of Galpha subunits and act in a G-protein-coupled receptor (GPCR)-specific manner. The conserved RGS domain accelerates the G subunit GTPase activity, whereas the variable amino-terminal domain participates in GPCR recognition. How receptor recognition is achieved is not known. Here, we show that the scaffold protein spinophilin (SPL), which binds the third intracellular loop (3iL) of several GPCRs, binds the N-terminal domain of RGS2. SPL also binds RGS1, RGS4, RGS16 and GAIP. When expressed in Xenopus laevis oocytes, SPL markedly increased inhibition of alpha-adrenergic receptor (alphaAR) Ca2+ signalling by RGS2. Notably, the constitutively active mutant alphaAR(A293E) (the mutation being in the 3iL) did not bind SPL and was relatively resistant to inhibition by RGS2. Use of betaAR-alphaAR chimaeras identified the 288REKKAA293 sequence as essential for the binding of SPL and inhibition of Ca2+ signalling by RGS2. Furthermore, alphaAR-evoked Ca2+ signalling is less sensitive to inhibition by SPL in rgs2-/- cells and less sensitive to inhibition by RGS2 in spl-/- cells. These findings provide a general mechanism by which RGS proteins recognize GPCRs to confer signalling specificity. PMID- 15793569 TI - Regulation of PTEN by Rho small GTPases. AB - PTEN (phosphatase and tensin homologue) is a phosphatase that dephosphorylates both protein and phosphoinositide substrates. It is mutated in a variety of human tumours and has important roles in a diverse range of biological processes, including cell migration and chemotaxis. PTEN's intracellular localization and presumably activity are regulated by chemoattractants in Dictyostelium and mouse neutrophils. However, the mechanisms for its regulation remain elusive. Here we show that RhoA and Cdc42, members of the Rho family of small GTPases, regulate the intracellular localization of PTEN in leukocytes and human transfected embryonic kidney cells. In addition, active RhoA is able to stimulate the phospholipid phosphatase activity of PTEN in human embryonic kidney cells and leukocytes, and this regulation seems to require RhoA's downstream effector, RhoA associated kinase (Rock). Furthermore, we have identified key residues on PTEN that are required for its regulation by the small GTPase, and show that small GTPase-mediated regulation of PTEN has a significant role in the regulation of chemotaxis. PMID- 15793570 TI - An alpha4 integrin-paxillin-Arf-GAP complex restricts Rac activation to the leading edge of migrating cells. AB - Formation of a stable lamellipodium at the front of migrating cells requires localization of Rac activation to the leading edge. Restriction of alpha4 integrin phosphorylation to the leading edge limits the interaction of alpha4 with paxillin to the sides and rear of a migrating cell. The alpha4-paxillin complex inhibits stable lamellipodia, thus confining lamellipod formation to the cell anterior. Here we report that binding of paxillin to the alpha4 integrin subunit inhibits adhesion-dependent lamellipodium formation by blocking Rac activation. The paxillin LD4 domain mediates this reduction in Rac activity by recruiting an ADP-ribosylation factor GTPase-activating protein (Arf-GAP) that decreases Arf activity, thereby inhibiting Rac. Finally, the localized formation of the alpha4-paxillin-Arf-GAP complex mediates the polarization of Rac activity and promotes directional cell migration. These findings establish a mechanism for the spatial localization of Rac activity to enhance cell migration. PMID- 15793571 TI - Prediction and validation of the distinct dynamics of transient and sustained ERK activation. AB - To elucidate the hidden dynamics of extracellular-signal-regulated kinase (ERK) signalling networks, we developed a simulation model of ERK signalling networks by constraining in silico dynamics based on in vivo dynamics in PC12 cells. We predicted and validated that transient ERK activation depends on rapid increases of epidermal growth factor and nerve growth factor (NGF) but not on their final concentrations, whereas sustained ERK activation depends on the final concentration of NGF but not on the temporal rate of increase. These ERK dynamics depend on Ras and Rap1 dynamics, the inactivation processes of which are growth factor-dependent and -independent, respectively. Therefore, the Ras and Rap1 systems capture the temporal rate and concentration of growth factors, and encode these distinct physical properties into transient and sustained ERK activation, respectively. PMID- 15793572 TI - Spin chirality on a two-dimensional frustrated lattice. AB - The collective behaviour of interacting magnetic moments can be strongly influenced by the topology of the underlying lattice. In geometrically frustrated spin systems, interesting chiral correlations may develop that are related to the spin arrangement on triangular plaquettes. We report a study of the spin chirality on a two-dimensional geometrically frustrated lattice. Our new chemical synthesis methods allow us to produce large single-crystal samples of KFe3(OH)6(SO4)2, an ideal Kagome lattice antiferromagnet. Combined thermodynamic and neutron scattering measurements reveal that the phase transition to the ordered ground-state is unusual. At low temperatures, application of a magnetic field induces a transition between states with different non-trivial spin textures. PMID- 15793573 TI - Improving the nutritional value of Golden Rice through increased pro-vitamin A content. AB - "Golden Rice" is a variety of rice engineered to produce beta-carotene (pro vitamin A) to help combat vitamin A deficiency, and it has been predicted that its contribution to alleviating vitamin A deficiency would be substantially improved through even higher beta-carotene content. We hypothesized that the daffodil gene encoding phytoene synthase (psy), one of the two genes used to develop Golden Rice, was the limiting step in beta-carotene accumulation. Through systematic testing of other plant psys, we identified a psy from maize that substantially increased carotenoid accumulation in a model plant system. We went on to develop "Golden Rice 2" introducing this psy in combination with the Erwinia uredovora carotene desaturase (crtI) used to generate the original Golden Rice. We observed an increase in total carotenoids of up to 23-fold (maximum 37 microg/g) compared to the original Golden Rice and a preferential accumulation of beta-carotene. PMID- 15793574 TI - Infrared spectroscopic imaging for histopathologic recognition. AB - The process of histopathology, comprising tissue staining and morphological pattern recognition, has remained largely unchanged for over 140 years. Although it is integral to clinical and research activities, histopathologic recognition remains a time-consuming, subjective process to which only limited statistical confidence can be assigned because of inherent operator variability. Although immunohistochemical approaches allow limited molecular detection, significant challenges remain in using them for quantitative, automated pathology. Vibrational spectroscopic approaches, by contrast, directly provide nonperturbing molecular descriptors, but a practical spectroscopic protocol for histopathology is lacking. Here we couple high-throughput Fourier transform infrared (FTIR) spectroscopic imaging of tissue microarrays with statistical pattern recognition of spectra indicative of endogenous molecular composition and demonstrate histopathologic characterization of prostatic tissue. This automated histologic segmentation is applied to routine archival tissue samples, incorporates well defined tests of statistical significance and eliminates any requirement for dyes or molecular probes. Finally, we differentiate benign from malignant prostatic epithelium by spectroscopic analyses. PMID- 15793575 TI - TIM-1 induces T cell activation and inhibits the development of peripheral tolerance. AB - We have examined the function of TIM-1, encoded by a gene identified as an 'atopy susceptibility gene' (Havcr1*), and demonstrate here that TIM-1 is a molecule that costimulates T cell activation. TIM-1 was expressed on CD4(+) T cells after activation and its expression was sustained preferentially in T helper type 2 (T(H)2) but not T(H)1 cells. In vitro stimulation of CD4(+) T cells with a TIM-1 specific monoclonal antibody and T cell receptor ligation enhanced T cell proliferation; in T(H)2 cells, such costimulation greatly enhanced synthesis of interleukin 4 but not interferon-gamma. In vivo, the use of antibody to TIM-1 plus antigen substantially increased production of both interleukin 4 and interferon-gamma in unpolarized T cells, prevented the development of respiratory tolerance, and increased pulmonary inflammation. Our studies suggest that immunotherapies that regulate TIM-1 function may downmodulate allergic inflammatory diseases. PMID- 15793576 TI - TIM-4 is the ligand for TIM-1, and the TIM-1-TIM-4 interaction regulates T cell proliferation. AB - The newly identified TIM family of proteins is associated with regulation of T helper type 1 (T(H)1) and T(H)2 immune responses. TIM-1 is genetically linked to asthma and is a receptor for hepatitis A virus, but the endogenous ligand of TIM 1 is not known. Here we show that TIM-4, which is expressed by antigen-presenting cells, is the ligand for TIM-1. In vivo administration of either soluble TIM-1 immunoglobulin (TIM-1-Ig) fusion protein or TIM-4-Ig fusion protein resulted in hyperproliferation of T cells, and TIM-4-Ig costimulated T cell proliferation mediated by CD3 and CD28 in vitro. These data suggest that the TIM-1-TIM-4 interaction is involved in regulating T cell proliferation. PMID- 15793577 TI - The subthalamic nucleus exerts opposite control on cocaine and 'natural' rewards. AB - A challenge in treating drug addicts is preventing their pathological motivation for the drug without impairing their general affective state toward natural reinforcers. Here we have shown that discrete lesions of the subthalamic nucleus greatly decreased the motivation of rats for cocaine while increasing it for food reward. The subthalamic nucleus, a key structure controlling basal ganglia outputs, is therefore able to oppositely modulate the effect of 'natural' rewards and drugs of abuse on behavior. Modulating the activity of the subthalamic nucleus might prove to be a new target for the treatment of cocaine addiction. PMID- 15793578 TI - Flexible strategies for sensory integration during motor planning. AB - When planning target-directed reaching movements, human subjects combine visual and proprioceptive feedback to form two estimates of the arm's position: one to plan the reach direction, and another to convert that direction into a motor command. These position estimates are based on the same sensory signals but rely on different combinations of visual and proprioceptive input, suggesting that the brain weights sensory inputs differently depending on the computation being performed. Here we show that the relative weighting of vision and proprioception depends both on the sensory modality of the target and on the information content of the visual feedback, and that these factors affect the two stages of planning independently. The observed diversity of weightings demonstrates the flexibility of sensory integration and suggests a unifying principle by which the brain chooses sensory inputs so as to minimize errors arising from the transformation of sensory signals between coordinate frames. PMID- 15793580 TI - A chimeric human-cat fusion protein blocks cat-induced allergy. AB - Animal allergens are an important cause of asthma and allergic rhinitis. We designed and tested a chimeric human-cat fusion protein composed of a truncated human IgG Fcgamma1 and the major cat allergen Fel d1, as a proof of concept for a new approach to allergy immunotherapy. This Fcgamma-Fel d1 protein induced dose dependent inhibition of Fel d1-driven IgE-mediated histamine release from cat allergic donors' basophils and sensitized human cord blood-derived mast cells. Such inhibition was associated with altered Syk and ERK signaling. The Fcgamma Fel d1 protein also blocked in vivo reactivity in FcepsilonRIalpha transgenic mice passively sensitized with human IgE antibody to cat and in Balb/c mice actively sensitized against Fel d1. The Fcgamma-Fel d1 protein alone did not induce mediator release. Chimeric human Fcgamma-allergen fusion proteins may provide a new therapeutic platform for the immune-based therapy of allergic disease. PMID- 15793579 TI - High cholesterol level is essential for myelin membrane growth. AB - Cholesterol in the mammalian brain is a risk factor for certain neurodegenerative diseases, raising the question of its normal function. In the mature brain, the highest cholesterol content is found in myelin. We therefore created mice that lack the ability to synthesize cholesterol in myelin-forming oligodendrocytes. Mutant oligodendrocytes survived, but CNS myelination was severely perturbed, and mutant mice showed ataxia and tremor. CNS myelination continued at a reduced rate for many months, and during this period, the cholesterol-deficient oligodendrocytes actively enriched cholesterol and assembled myelin with >70% of the cholesterol content of wild-type myelin. This shows that cholesterol is an indispensable component of myelin membranes and that cholesterol availability in oligodendrocytes is a rate-limiting factor for brain maturation. PMID- 15793581 TI - Kielin/chordin-like protein, a novel enhancer of BMP signaling, attenuates renal fibrotic disease. AB - The bone morphogenetic proteins (BMPs) profoundly affect embryonic development, differentiation and disease. BMP signaling is suppressed by cysteine-rich domain proteins, such as chordin, that sequester ligands from the BMP receptor. We describe a novel protein, KCP, with 18 cysteine-rich domains. Unlike chordin, KCP enhances BMP signaling in a paracrine manner. Smad1-dependent transcription and phosphorylated Smad1 (P-Smad1) levels are increased, as KCP binds to BMP7 and enhances binding to the type I receptor. In vivo, Kcp(-/-) mice are viable and fertile. Because BMPs have a pivotal role in renal disease, we examined the phenotype of Kcp(-/-) mice in two different models of renal injury. Kcp(-/-) animals show reduced levels of P-Smad1, are more susceptible to developing renal interstitial fibrosis, are more sensitive to tubular injury and show substantial pathology after recovery. The data indicate an important role for KCP in attenuating the pathology of renal fibrotic disease. PMID- 15793582 TI - Calmodulin kinase II inhibition protects against structural heart disease. AB - Beta-adrenergic receptor (betaAR) stimulation increases cytosolic Ca(2+) to physiologically augment cardiac contraction, whereas excessive betaAR activation causes adverse cardiac remodeling, including myocardial hypertrophy, dilation and dysfunction, in individuals with myocardial infarction. The Ca(2+)-calmodulin dependent protein kinase II (CaMKII) is a recently identified downstream element of the betaAR-initiated signaling cascade that is linked to pathological myocardial remodeling and to regulation of key proteins involved in cardiac excitation-contraction coupling. We developed a genetic mouse model of cardiac CaMKII inhibition to test the role of CaMKII in betaAR signaling in vivo. Here we show CaMKII inhibition substantially prevented maladaptive remodeling from excessive betaAR stimulation and myocardial infarction, and induced balanced changes in excitation-contraction coupling that preserved baseline and betaAR stimulated physiological increases in cardiac function. These findings mark CaMKII as a determinant of clinically important heart disease phenotypes, and suggest CaMKII inhibition can be a highly selective approach for targeting adverse myocardial remodeling linked to betaAR signaling. PMID- 15793583 TI - Apolipoprotein M is required for prebeta-HDL formation and cholesterol efflux to HDL and protects against atherosclerosis. AB - High-density lipoproteins (HDLs) are considered antiatherogenic because they mediate reverse cholesterol transport from the periphery to the liver for excretion and degradation. Here we show that mice deficient in apolipoprotein M (apoM), a component of the HDL particle, accumulated cholesterol in large HDL particles (HDL(1)) while the conversion of HDL to prebeta-HDL was impaired. Accordingly, apoM-deficient mice lacked prebeta-HDL, a subclass of lipid-poor apolipoproteins that serves as a key acceptor of peripheral cellular cholesterol. This deficiency led to a markedly reduced cholesterol efflux from macrophages to apoM-deficient HDL compared to normal HDL in vitro. Overexpression of apoM in Ldlr(-/-) mice protected against atherosclerosis when the mice were challenged with a cholesterol-enriched diet, showing that apoM is important for the formation of prebeta-HDL and cholesterol efflux to HDL, and thereby inhibits formation of atherosclerotic lesions. PMID- 15793584 TI - A highly conserved 6S RNA structure is required for regulation of transcription. AB - 6S RNA, a highly abundant noncoding RNA, regulates transcription through interaction with RNA polymerase in Escherichia coli. Computer searches identified 6S RNAs widely among gamma-proteobacteria. Biochemical approaches were required to identify more divergent 6S RNAs. Two Bacillus subtilis RNAs were found to interact with the housekeeping form of RNA polymerase, thereby establishing them as 6S RNAs. A third B. subtilis RNA was discovered with distinct RNA polymerase binding activity. Phylogenetic comparison and analysis of mutant RNAs revealed that a conserved secondary structure containing a single-stranded central bulge within a highly double-stranded molecule was essential for 6S RNA function in vivo and in vitro. Reconstitution experiments established the marked specificity of 6S RNA interactions for sigma(70)-RNA polymerase, as well as the ability of 6S RNA to directly inhibit transcription. These data highlight the critical importance of structural characteristics for 6S RNA activity. PMID- 15793586 TI - A centrosomal mechanism involving CDK5RAP2 and CENPJ controls brain size. AB - Autosomal recessive primary microcephaly is a potential model in which to research genes involved in human brain growth. We show that two forms of the disorder result from homozygous mutations in the genes CDK5RAP2 and CENPJ. We found neuroepithelial expression of the genes during prenatal neurogenesis and protein localization to the spindle poles of mitotic cells, suggesting that a centrosomal mechanism controls neuron number in the developing mammalian brain. PMID- 15793585 TI - Insights into hRPA32 C-terminal domain--mediated assembly of the simian virus 40 replisome. AB - Simian virus 40 (SV40) provides a model system for the study of eukaryotic DNA replication, in which the viral protein, large T antigen (Tag), marshals human proteins to replicate the viral minichromosome. SV40 replication requires interaction of Tag with the host single-stranded DNA-binding protein, replication protein A (hRPA). The C-terminal domain of the hRPA32 subunit (RPA32C) facilitates initiation of replication, but whether it interacts with Tag is not known. Affinity chromatography and NMR revealed physical interaction between hRPA32C and the Tag origin DNA-binding domain, and a structural model of the complex was determined. Point mutations were then designed to reverse charges in the binding sites, resulting in substantially reduced binding affinity. Corresponding mutations introduced into intact hRPA impaired initiation of replication and primosome activity, implying that this interaction has a critical role in assembly and progression of the SV40 replisome. PMID- 15793587 TI - Chfr is required for tumor suppression and Aurora A regulation. AB - Tumorigenesis is a consequence of loss of tumor suppressors and activation of oncogenes. Expression of the mitotic checkpoint protein Chfr is lost in 20-50% of primary tumors and tumor cell lines. To explore whether downregulation of Chfr contributes directly to tumorigenesis, we generated Chfr knockout mice. Chfr deficient mice are cancer-prone, develop spontaneous tumors and have increased skin tumor incidence after treatment with dimethylbenz(a)anthracene. Chfr deficiency leads to chromosomal instability in embryonic fibroblasts and regulates the mitotic kinase Aurora A, which is frequently upregulated in a variety of tumors. Chfr physically interacts with Aurora A and ubiquitinates Aurora A both in vitro and in vivo. Collectively, our data suggest that Chfr is a tumor suppressor and ensures chromosomal stability by controlling the expression levels of key mitotic proteins such as Aurora A. PMID- 15793588 TI - Genome-wide strategies for detecting multiple loci that influence complex diseases. AB - After nearly 10 years of intense academic and commercial research effort, large genome-wide association studies for common complex diseases are now imminent. Although these conditions involve a complex relationship between genotype and phenotype, including interactions between unlinked loci, the prevailing strategies for analysis of such studies focus on the locus-by-locus paradigm. Here we consider analytical methods that explicitly look for statistical interactions between loci. We show first that they are computationally feasible, even for studies of hundreds of thousands of loci, and second that even with a conservative correction for multiple testing, they can be more powerful than traditional analyses under a range of models for interlocus interactions. We also show that plausible variations across populations in allele frequencies among interacting loci can markedly affect the power to detect their marginal effects, which may account in part for the well-known difficulties in replicating association results. These results suggest that searching for interactions among genetic loci can be fruitfully incorporated into analysis strategies for genome wide association studies. PMID- 15793589 TI - Resveratrol rescues mutant polyglutamine cytotoxicity in nematode and mammalian neurons. AB - We report that Sir2 activation through increased sir-2.1 dosage or treatment with the sirtuin activator resveratrol specifically rescued early neuronal dysfunction phenotypes induced by mutant polyglutamines in transgenic Caenorhabditis elegans. These effects are dependent on daf-16 (Forkhead). Additionally, resveratrol rescued mutant polyglutamine-specific cell death in neuronal cells derived from HdhQ111 knock-in mice. We conclude that Sir2 activation may protect against mutant polyglutamines. PMID- 15793590 TI - Guide to receptors and channels, 1st edition (2005 revision). PMID- 15793601 TI - Right bundle branch block in acute myocardial infarction treated by primary coronary angioplasty and stenting. AB - Patients with right bundle branch block (RBBB) in acute myocardial infarction (AMI) have a significantly higher mortality rate even with the advent of thrombolytic therapy. This study was undertaken to assess the impact of primary percutaneous transluminal coronary angioplasty (PTCA) and stenting on the outcome of patients with RBBB in AMI. A total of 600 patients with AMI who underwent primary PTCA and stenting (rate: 61%) < 12 hours of onset were studied. A 12-lead ECG was obtained at least every 6 hours. Serial creatine kinase was measured, and left ventricular ejection fraction was obtained during the hospital stay. Among 600 patients with AMI, 94 patients (15.7%) had RBBB; it was persistent in 31 (33%) and transient in 63 (67%). In-hospital mortality rate was 7.3% in patients without RBBB, 7.9% in transient RBBB, and 25.8% in persistent RBBB (p < 0.02). The incidence of heart failure was 26.5% in those without RBBB, 34.9% in transient RBBB, and 58.1% in persistent RBBB (p < 0.001). There was no significant difference among these 3 groups in ventricular arrhythmias and complete atrioventricular block. Peak creatine kinase was 3,214+/-2,293 U/L in those without RBBB, 4,558+/-3,316 U/L in transient RBBB (p < 0.001), and 5,635+/ 3,920 U/L in persistent RBBB (p < 0.001). Left ventricular ejection fraction was 50+/-11% in those without RBBB, 47+/-11% in transient RBBB (p < 0.05), and 42+/ 13% in persistent RBBB (p < 0.001). Patients with AMI treated by primary PTCA and stenting had an increased incidence of transient RBBB, especially following reperfusion therapy, although the clinical outcome was similar to that of those without RBBB. In contrast, there was no satisfactory improvement in clinical outcomes in those with persistent RBBB. PMID- 15793600 TI - Management of cardiogenic shock due to acute coronary syndromes. AB - Despite advances in the treatment of patients with acute coronary syndromes, there has been no significant decrease in the incidence of cardiogenic shock, while its mortality remains frustratingly high. Shock is a progressive state of hypotension (systolic blood pressure < 90 mm Hg) lasting at least 30 minutes, which leads to systemic hypoperfusion. It is more common in patients with ST segment elevation myocardial infarction than in patients with other acute coronary syndromes. Revascularization is associated with better outcomes than intensive medical therapy, especially in patients < 75 years of age with cardiogenic shock. Adjunctive therapies include inotropes, vasopressor therapy, intra-aortic balloon pump counterpulsation, and IIb/IIIa blockade to prevent no reflow phenomenon during primary percutaneous transluminal coronary angioplasty. Other adjunctive therapies which are investigated are improved mechanical support devices, and as medical therapy for myocyte protection nicorandil, glucose/insulin/potassium infusions and direct inhibition of Na+/H+ exchanger. PMID- 15793602 TI - Quantitative angiographic analysis of right ventricular global function in normal incidences and in patients with coronary artery disease. AB - At present right ventriculography data cannot be accurately estimated owing to the absence of software for quantitative analysis of the right ventricle (RV) volumes and function. The aim of this study was to use existing software for left ventriculography analysis to estimate right ventricular volumes and function in patients without coronary lesions and in those with coronary artery disease (CAD). Thirty-two patients without significant lesions of coronary arteries and 20 patients with CAD were examined with left ventriculography and right atriography. Each examination was performed in 2 projections: 30 degrees right anterior oblique (RAO 30 degrees) and 60 degrees left anterior oblique (LAO 60 degrees) projections. Correction factor (CF) was obtained by finding the ratio between stroke volumes of the left and right ventricles. The mean CF was 0.9243+/ 0.2887 for patients without CAD and 0.8758+/-0.2232 for patients with CAD. Such calculation is the easiest and quite accurate method of determining RV volumes and function by using existing software of quantitative angiographic analysis. PMID- 15793603 TI - Effect of direct stent implantation on QTc dispersion. AB - The aim of this study was to evaluate whether direct stenting is superior to conventional stent implantation technique with respect to QTc dispersion in prospectively selected patients with simple lesion morphology and class II stable angina undergoing elective coronary stenting. One hundred thirty-four consecutive patients were divided into 2 groups based on the stenting technique used: the direct stenting without predilation group, group I (n = 64), and the stenting with predilation group, group II (n = 70). All patients had single-vessel disease. The primary end point of the study was the QTc dispersion at the 24th hour and at the first month after the procedure and the secondary end point of the study was the major clinical events (MCEs) rate in the hospital period and up to 1 month. Baseline maximum QTc, minimum QTc, and QTc dispersion were not different between the 2 groups. QTc dispersion decreased from 47+/-8 msec before stent implantation to 41+/-11 msec at 24 hours and 37+/-7 msec 1 month after angioplasty in group I (p < 0.006 and p < 0.01, respectively), whereas QTc dispersion decreased from 49+/-9 msec before stent implantation to 46+/-8 msec at 24 hours and 42+/-10 msec 1 month after angioplasty in group II (p < 0.03 and p < 0.01, respectively). Compared with group II, the decrease in QTc dispersion was significantly greater at the 24th hour and at the first month after the procedure in group I (p < 0.003 and p < 0.001, respectively). There was a decreased trend toward MCE rate in group I in relation to that of group II, but the statistical difference was not significant. Direct stenting is a feasible and safe technique. It is superior to conventional stenting technique in decreasing the QTc dispersion at the 24th hour and at the first month after the procedure, whereas it is equivalent to single-vessel conventional stent implantation technique with respect to MCEs rate in the short-term period. PMID- 15793605 TI - Clinical, echocardiographic, and hemodynamic characteristics of rheumatic mitral valve stenosis and atrial fibrillation. AB - Chronic atrial fibrillation (AF) is associated with an increased frequency of embolic events and negative impact on cardiac function, and therefore, an increased morbidity and mortality risk in patients with rheumatic mitral valve stenosis (RMS). In the present study, the clinical, 2-D and Doppler echocardiographic, and left-and right-heart hemodynamic data were evaluated for 92 patients (68 women) with RMS and AF and compared with data from 118 patients (88 women) with RMS with sinus rhythm. The clinical, echocardiographic, and hemodynamic evaluations were performed within 1 to 7 days of each other. Patients with AF were older (45.7+/-13.4 vs 38.6+/-12.0 years, p < 0.01) and had a longer symptomatic period (108.2+/-117.9 vs 50.6+/-53.1 months, p < 0.01) compared with those with sinus rhythm. Most of the patients with AF were in NYHA functional capacity 3-4 (74% vs 19%), whereas most of the patients with sinus rhythm were in NYHA functional capacity 2. Patients with AF had a higher mitral valve score based on morphologic features ranging from 4 to 16 depending on the severity of disease (8.3+/-2.1 vs 6.5+/-1.9, p < 0.01) and greater left ventricular end diastolic diameter (LVEDD) (52.3+/-8.7 vs 47.7+/-8.7 mm, p < 0.02), and end systolic diameter (LVESD) (34.4+/-7.5 vs 30.9+/-7.5 mm, p < 0.01). Organic tricuspid valve involvement was diagnosed more frequently in patients with AF (61% vs 32%, p < 0.01). Mild mitral regurgitation was also more frequent in patients with AF (71%vs 51%, p < 0.03). The mitral valve area was similar in patients with and without AF (1.30+/-0.39 vs 1.39+/-0.41 cm2, p > 0.05). Mean diastolic mitral valve gradient and pulmonary artery pressure did not differ in patients with and without AF. Right atrial pressures were higher in patients with AF (7.6+/-3.3 vs 6.3+/-1.9 mm Hg, p < 0.02). The authors suggest that (1) AF occurred in older patients, who had a longer disease process and more serious symptoms; (2) hemodynamic derangements (mitral valve gradient, pulmonary artery pressure) did not differ in patients with and without AF; (3) greater mitral valve score, more tricuspid valve involvement, higher LVEDD, which are suggestive of greater rheumatic activity process were more frequently seen in patients with AF than in those without AF. These findings support the opinion that AF is a marker of widespread rheumatic damage in patients with RMS. PMID- 15793604 TI - Safety and efficacy of enalapril in multivalvular heart disease with significant mitral stenosis--SCOPE-MS. AB - Angiotensin-converting enzyme inhibitors (ACEI) are often used in preventing and treating heart failure due to regurgitant valve disease. The majority of patients with symptomatic rheumatic heart disease (RHD) have significant mitral stenosis (MS) and are denied ACEI therapy, because of the fear of hypotension in the presence of fixed obstruction. The authors assessed the safety and efficacy of ACEI in 109 consecutive patients with RHD and with significant mitral stenosis (mitral valve orifice, MVO < 1.5 cm2)and with NYHA class III or IV heart failure symptoms. Mean age was 33.1+/-12 years, systolic blood pressure (BP) was 111+/ 10, and diastolic BP was 73+/-8 mm Hg. MS was significant in 100 patients with mitral regurgitation in 46, aortic regurgitation in 19, and pulmonary hypertension in 60 patients. After initial stabilization, enalapril 2.5 mg bid was started in hospital and titrated up to 10 mg bid over 2 weeks. NYHA status, Borg score, and 6-minute walk test were assessed at baseline, and at 1, 2, and 4 weeks. Seventy-nine of the 100 patients who completed the study had severe MS (MVO < 1.0 cm2). Enalapril was well tolerated by all study patients without hypotension or worsening of symptoms. NYHA class (3.2+/-0.5 baseline vs 2.3+/-0.5 at 4 weeks, p < 0.01) Borg Dyspnea Index (7.6+/-1.3 vs 5.6+/-1.3, p < 0.01), and 6-minute walk distance (226+/-106 vs 299+/-127 m, p < 0.01) improved significantly with enalapril. Patients with associated regurgitant lesions showed more improvement in exercise capacity (120+/-93 vs 39+/-56 m, p < 0.001). Enalapril was well tolerated in patients with RHD with moderate and severe MS. Irrespective of the valve pathology, enalapril improved functional status and exercise capacity with maximum benefit in patients with concomitant regurgitant valvular heart disease. PMID- 15793607 TI - Matrix metalloproteinases and diabetic vascular complications. AB - Diabetes mellitus (DM) is associated with an increased incidence of cardiovascular events and microvascular complications. These complications contribute to the morbidity and mortality associated with DM. There is increasing evidence supporting a role for matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of matrix metalloproteinases - TIMPs) in the atherosclerotic process. However, the relationship between MMPs/TIMPs and diabetic angiopathy is less well defined. Hyperglycemia directly or indirectly (eg, via oxidative stress or advanced glycation products) increases MMP expression and activity. These changes are associated with histologic alterations in large vessels. On the other hand, low proteolytic activity of MMPs contributes to diabetic nephropathy. Within atherosclerotic plaques an imbalance between MMPs and TIMPs may induce matrix degradation, resulting in an increased risk of plaque rupture. Furthermore, because MMPs enhance blood coagulability, MMPs and TIMPs may play a role in acute thrombotic occlusion of vessels and consequent cardiovascular events. Some drugs can inhibit MMP activity. However, the precise mechanisms involved are still not defined. Further research is required to demonstrate the causative relationship between MMPs/TIMPs and diabetic atherosclerosis. It also remains to be established if the long-term administration of MMP inhibitors can prevent acute cardiovascular events. PMID- 15793606 TI - HR, 0-(beta-hydroxyethyl)-rutosides; (Venoruton): rapid relief of signs/symptoms in chronic venous insufficiency and microangiopathy: a prospective, controlled study. AB - The aim of this independent study was to demonstrate the rapidity of the clinical action of HR 0-(beta-hydroxyethyl)-rutosides, Venoruton (Novartis Consumer Health) in patients with chronic venous insufficiency (CVI). Two groups of patients with venous hypertension and microangiopathy were treated with HR (1 or 2 g/day, for 8 weeks). Twelve patients (age 56.4; range 44-66; M:F = 6:6) were included in group 1 (1 g/day) (moderate CVI and microangiopathy); 10 patients (age 57.4; range 42-67; M:F = 5:5) in group 2 (2 g/day) with more severe CVI and microangiopathy. Average ambulatory venous pressure (AVP) was 58.6 (range 50-65) with a refilling time (RT) shorter than 10 seconds. There were no significant differences in AVP and RT between the 2 groups, but the duration of the disease was longer in group 2: 3.5 years (SD 2.0) in group 1 and 6.4 years (SD 3.3) in group 2. All included subjects completed the study and no dropouts were observed. In both dose groups there was a progressive decrease in laser Doppler resting flux (RF), indicating improvement in microangiopathy and a significant decrease in capillary filtration (RAS) associated with a significant improvement in analogue scale line score (ASLS) and edema. Although the effect in the 2 g dose group was more rapid on the microcirculatory parameters with a significant effect on RF and RAS after 4 days (effect of 1 g per day after 8 days and 6 days, respectively), there was no difference in the time to onset of a significant clinical improvement (ie, the ASLS and the edema score): 4 days in both groups. Venous microangiopathy and edema were improved by the treatment with HR within a few days. The effects were visible with both dosages, in both severity groups. PMID- 15793608 TI - Effect of cholesterol lowering and cardiovascular risk factors on the progression of aortoiliac arteriosclerosis: a quantitative cineangiography study. AB - The post-Coronary Artery Bypass Graft (Post-CABG) trial has shown that aggressive compared to moderate lowering of low-density lipoprotein cholesterol (LDL-C) delayed the progression of obstructive disease in aortocoronary saphenous vein grafts and in the left main coronary artery. Patients had been allocated to high and low-dose lovastatin therapy for a 4-5 year period. The present study evaluated the effect of LDL-C lowering and the role of cardiovascular risk factors on the progression of arteriosclerosis in the distal abdominal aorta and common iliac arteries. From one of the participating centers of the post-CABG trial, 145 patients who had adequate imaging of the aortoiliac arteries at baseline and follow-up were included. Angiographic outcomes, presumed to reflect progression of arteriosclerosis and obtained from lumen diameter (LD) measurements using quantitative cineangiography, were as follows: significant decrease of the minimum lumen diameter (LD) and increase of the maximum LD, percent lumen stenosis, and percent lumen dilatation. These outcomes were not significantly less frequent in patients randomly allocated to aggressive compared to moderate LDL-C lowering. Of 9 cardiovascular risk factors, only 2 were significantly related to progression of aortoiliac arteriosclerosis. Current smoking predicted both percent lumen stenosis increase and, to a lesser degree, percent lumen dilatation increase (p = 0.010 and p = 0.055, respectively). Abnormally high body mass index (BMI > or = 25 kg/m2) correlated with percent lumen dilatation increase (p = 0.006). Aggressive compared to moderate LDL-C lowering did not prevent or delay the progression of aortoiliac arteriosclerosis. Smoking predicted both lumen narrowing and dilatation presumably caused by arteriosclerosis. Abnormally high BMI, reflecting overweight or obesity, was strongly associated with vessel dilatation. PMID- 15793609 TI - Fractions of total plasma homocysteine in patients with ischemic stroke before the age of 55 years. AB - The mechanism responsible for the association between elevated circulating homocysteine levels and ischemic stroke remains unclear. Therefore, the authors assessed total plasma homocysteine (tHcy) and its fractions (free [fHcy] and protein-bound [bHcy] homocysteine) in patients with ischemic stroke before the age of 55 years. Fifty patients (23 men, mean age 46.8+/-7.6 years) with ischemic stroke or transient ischemic attacks, with symptoms lasting < 72 hours were enrolled. In this group: 32 (64%) patients had hypertension; 12 (24%), ischemic heart disease (IHD); and 20 (40%), type 2 diabetes mellitus (DM). The control group consisted of 30 matched healthy individuals (17 men, mean age 44.6+/-6.2 years). The tHcy, fHcy, and bHcy levels were determined by high-performance liquid chromatography. tHcy and its fractions did not differ significantly between patients and controls. However, stroke patients with hypertension had significantly higher concentrations of tHcy and bHcy compared to stroke patients without hypertension (tHcy 13.0+/-3.3 vs 10.7 +/-3.2 micromol/L, p < 0.05; bHcy 9.7+/-2.6 vs 7.8+/-2.3 micromol/L, p < 0.01, respectively); fHcy was borderline significant: 3.1 (1.5-6.5) vs 2.5 (1.8-5.3) micromol/L, p = 0.05. The presence of IHD, DM, hyperlipoproteinemia, clinical subtypes of stroke, smoking, and family history of stroke did not influence these parameters. In the group of 50 patients, tHcy correlated with mean systolic blood pressure (BP) (r = 0.3, p < 0.05) and bHcy correlated with mean systolic and mean diastolic BP (r = 0.3, p < 0.05). These findings suggest an association between hypertension and redox status of Hcy in patients with ischemic stroke before the age of 55 years. This observation supports the hypothesis that elevated BP may contribute to Hcy related vascular injury. PMID- 15793610 TI - Provocation of microvessel spasm by low-dose acetylcholine in patients with suspected coronary artery disease--two case reports. AB - Endothelial dysfunction plays an important role in the pathogenesis of cardiac syndrome X, and intracoronary low-dose acetylcholine infusion is a widely used diagnostic modality for studying the coronary artery endothelial function. The authors herein report 2 cases of cardiac syndrome X with coronary artery endothelial dysfunction and microvessel spasm. The findings of non-invasive testing were positive for ischemia. Coronary angiograms appeared entirely normal in both cases. However, the intracoronary infusion of low-dose (1.5-15 microg/minute) acetylcholine demonstrated an impairment of the coronary blood flow response and consequently provoked an ST-segment elevation in an electrocardiogram. The coronary angiograms showed no spasm in the epicardial arteries. These patients are thus suggested to have cardiac syndrome X with microvessel spasms associated with coronary artery endothelial dysfunction. PMID- 15793611 TI - Hyperthyroidism and complete atrioventricular block--a report of 2 cases with electrophysiologic assessment. AB - Although cardiovascular manifestations in thyroid disorders are frequently encountered in clinical practice, atrioventricular (AV) conduction disorders, especially in hyperthyroidism, are rare. There are some proposed mechanisms for AV blocks in hyperthyroidism but the exact mechanism is still unknown. The authors report 2 cases with thyroid function disorders and complete AV block, and the electrophysiologic characteristics of these 2 patients, and they review and speculate on similar reported cases. PMID- 15793612 TI - Radiofrequency catheter ablation of accessory pathway in a patient with Ebstein's anomaly and atrial septal defect--a case report. AB - The authors report the case of a 57-year-old woman with Ebstein's anomaly and atrial septal defect. She was referred to their hospital for treatment of refractory paroxysmal wide QRS tachycardia. Her 12-leads ECG in sinus rhythm showed ventricular preexcitation of type B Wolff-Parkinson-White syndrome. In a baseline electrophysiological study, a wide QRS tachycardia with right bundle branch block configurations was induced. This tachycardia was orthodromic atrioventricular reciprocating tachycardia with a right inferior accessory pathway. Radiofrequency current was successfully delivered at the inferior site of the atrialized right ventricle. Radiofrequency catheter ablation seems to be useful for supraventricular tachycardia in patients with Ebstein's anomaly and atrial septal defect. PMID- 15793613 TI - Severe aortic regurgitation in von Recklinghausen's disease--a case report. AB - A 71-year-old man with von Recklinghausen's disease presented with NYHA class II exertional dyspnea and easy fatigability since about 20 days. Echocardiography and angiography demonstrated dilatation of aortic root, ascending aorta, and left ventricle, together with severe aortic regurgitation. Because of very few cases of cardiovascular abnormalities having been reported with von Recklinghausen's disease, it is logical to concur that this is simply co-existence, whereas other possible mechanisms are endothelial dysfunction or sympathetic denervation. PMID- 15793614 TI - Thrombocytopenia, immunoglobulin treatment, and acute myocardial infarction--a case report. AB - Platelets play a pivotal role in the pathophysiology of the acute coronary syndromes, and platelet inhibition is a cornerstone in the management of these patients. Patients with profound thrombocytopenia who present with an acute coronary syndrome present a difficult challenge. The authors report a patient with immune thrombocytopenic purpura who presented with acute myocardial infarction despite a very low platelet count and who sustained recurrent infarction after receiving immune globulin treatment. The best management of thrombocytopenic patients with acute coronary syndromes is uncertain, but extreme caution is needed before efforts are made to raise the platelet count in order to allow conventional treatment. PMID- 15793615 TI - Large saccular aneurysm in a coronary arterial fistula--a case report. AB - The authors present a case of a large saccular aneurysm in a coronary artery fistula originating from the proximal left anterior descending coronary artery, draining into the main trunk of the pulmonary artery. The diagnosis was made by 3 dimensional computed tomography and coronary arteriography. Congenital coronary artery fistula is not uncommon. With a saccular aneurysm, however, it is very rare. A ruptured aneurysm will induce sudden death if surgical repair is not done. PMID- 15793616 TI - GRAFT infection of thoracic aorta due to group C beta-hemolytic streptococcus--a case report. AB - A fatal case of late-onset graft infection of the thoracic aorta due to group C beta-hemolytic streptococcus is described. A 37-year-old male patient, who had a history of total aortic arch replacement for acute aortic dissection 8 years before, was admitted to the department. He suffered from toxic shock syndrome, disseminated intravascular coagulation (DIC), and acute renal failure. Group C beta-hemolytic streptococcus was detected from his blood; however, echocardiography, computed tomography (CT), and magnetic resonance imaging (MRI) failed to detect the focus of the infection. In spite of intensive care, including antibiotic therapy, artificial ventilation, and continuous hemodiafiltration, he died on the 18th day of hospitalization. Autopsy revealed that a small abscess was present at the proximal anastomotic segments of the patient's graft. A bite inflicted by his dog, 14 days before admission, was suspected to be the source of this bacterium. A rare case of graft infection of thoracic aorta in terms of causative organism, long period from graft replacement to graft infection, and site of infection is presented and discussed. PMID- 15793617 TI - Isolation and characterization of alachlor-degrading actinomycetes from soil. AB - Alachlor (2-cloro-N-(methoxymethyl)-N-(2,6-diethylphenyl)-acetamide) is an extremely toxic and highly mobile herbicide that is widely used for pre-emergence control of grasses and weeds in many commercial crops in Brazil. In order to select soil actinomycetes able to degrade this herbicide, fifty-three actinomycete strains were isolated from soil treated with alachlor using selective conditions and subjected to in vitro degradation assays. Sixteen isolates were shown to be tolerant to high concentrations of the herbicide (up to 720 mg L(-1)), and six of these were able to grow and degrade >/= 50 alachlor (72 mg L(-1)) in mineral salts medium. Morphological and phylogenetic analysis enabled the assignment of the alachlor-degrading strains to the genus Streptomyces. Strain LS151 was related to the type strains of Streptomyces capoamus/Streptomyces galbus, whereas strains LS143 and LS153 were related to Streptomyces bikiniensis. The remaining strains, LS166, LS177 and LS182, were similar in morphological features and recovered in a single cluster based on 16S rDNA sequence analysis, but shown to be distinct on the basis of genomic fingerprint data (rep-PCR). Though a definitive taxonomic assignment of alachlor degrading strains was not possible, these data indicate that ability to degrade this pesticide was detected in different Streptomyces taxa. PMID- 15793618 TI - Expression of PHA polymerase genes of Pseudomonas putida in Escherichia coli and its effect on PHA formation. AB - Poly-3-hydroxyalkanoates (PHAs) are synthesized by many bacteria as intracellular storage material. The final step in PHA biosynthesis is catalyzed by two PHA polymerases (phaC) in Pseudomonas putida. The expression of these two phaC genes (phaC1 and phaC2)was studied in Escherichia coli, either under control of the native promoter or under control of an external promoter. It was found that the two phaC genes are not expressed in E. coli without an external promoter. During heterologous expression of phaC from Plac on a high copy number plasmid, a rapid reduction of the number of colony forming units was observed, especially for phaC2. It appears that the plasmid instability was partially caused by high-level production of PHA polymerase. Subsequently, tightly regulated phaC2 expression systems on a low copy number vector were applied in E. coli. This resulted in PHA yields of over 20 of total cell dry weight, which was 2 fold higher than that obtained from the system where phaC2 is present on a high copy number vector. In addition, the PHA monomer composition differed when different gene expression systems or different phaC genes were applied. PMID- 15793619 TI - Cryptococcus allantoinivorans sp.nov., an anamorphic basidiomycetous yeast (Tremellales) physiologically resembling other species of the Cryptococcus laurentii complex that degrade polysaccharides and C2 compounds. AB - A novel Cryptococcus species is proposed to accommodate a yeast strain (CBS 9604) able to assimilate allantoin as sole carbon source, a characteristic very uncommon among yeasts. By traditional methods, the strain could not be distinguished from Cryptococcus laurentii, but nucleotide sequences of the D1D2 region of the large subunit (26S) and of the ITS region of ribosomal DNA showed relationship to the Bulleromyces clade of the genus Cryptococcus (order Tremellales) with some Tremella spp. as the closest relatives. A traditional morphological and physiological description of the strain is given. Data on the assimilation of some C2 compounds and polysaccharides are provided and compared with those of other type strains of novel species of the C. laurentii complex. PMID- 15793620 TI - Fungal diversity from western redcedar fences and their resistance to beta thujaplicin. AB - The work reported here investigated the fungal community inhabiting western redcedar fence material with a focus on species colonizing wood below the surface, of which little is known. From seven pieces of fence material, twenty three different fungal species were isolated and characterized using both traditional morphology and molecular identification methods. The species identified included thirteen ascomycetous and ten basidiomycetous fungi. Isolates were tested for their resistance to beta-thujaplicin - one of the principle fungicidal agents of western redcedar heartwood extractives. Generally, ascomycetous fungi exhibited greater resistance to beta-thujaplicin than basidiomycetous fungi. Interestingly, three ascomycetous and two basidiomycetous species frequently isolated had high tolerance to this compound. These species could be candidate 'pioneer' species that invade and detoxify western redcedar extractives, paving the way for colonization by decay fungi. PMID- 15793621 TI - NAD deamidation "a new reaction" by an enzyme from Aspergillus terreus DSM 826. AB - NAD deamidation is a non-previously recognized reaction. This reaction has been found to be catalyzed by extracts of Aspergillus terreus DSM 826. Conversion of NAD to the biosynthetic intermediate, deamido NAD, by these extracts, at the optimum pH and temperature did not exceed about 55 of the amount of the substrate added. Completion of the reaction was achieved when the extracts were pre-heated at 50 degrees C for 15 min in absence of the substrate. In a very similar manner, the extracts catalyzed hydrolytic cleavage of the amide linkages of different biomolecules such as nicotinamide, nicotinamide riboside, nicotinamide mononucleotide, L-glutamine, L-asparagine and acetamide. Polyacrylamide was also deamidated under the same conditions. In addition, complete dephosphorylation of the dinucleotide molecule was also effected by the same extracts. Separation of the NAD deamidating enzyme from the NAD dephosphorylating enzyme was achieved on using either DEAE - Sephadex A-25 or Sephadex G-200 column chromatography. The obtained phosphohydrolase-free-deamidase showed optimum activity at pH 8 of 0.1 M phosphate buffer and 50 degrees C. It exhibited broad substrate specificity and hyperbolic substrate saturation kinetics. It was isosterically inhibited by the product of its activity and this inhibition was prevented by heating the extracts at 50 degrees C for 15 min. Its activity was not affected in presence of sodium fluoride, partially inhibited in presence of magnesium chloride and was retained in the freezer for some months. PMID- 15793622 TI - Uncovering the first double brimmed hat-shaped ascospores in Ambrosiozyma platypodis Van der Walt. AB - Using transmission electron microscopy with glutaraldehyde and osmium tetroxide as chemical fixatives, hat-shaped ascospores with two brims each were uncovered in the yeast Ambrosiozyma platypodis. This is the first report on such structures. PMID- 15793623 TI - Drug therapy in pediatric cardiology and the role of digoxin in pediatric congestive heart failure. PMID- 15793624 TI - Posterior descending coronary artery flow reserve assessment by Doppler echocardiography in children with and without congenital heart defect: comparison with invasive technique. AB - To evaluate whether transthoracic Doppler echocardiography can reliably measure coronary flow velocity and coronary flow velocity reserve (CFVR) in the posterior descending coronary artery (PD) in children, we examined 32 patients who had congenital heart disease (ventricular septal defect in 10, tetralogy of Fallot in 6, tricuspid atresia in 3, double-outlet right ventricle in 2, patent ductus arteriosus in 2, and aortic valve stenosis in 2) and 7 patients who had a history of Kawasaki disease without stenosis or aneurysm formation of the coronary artery. Average peak flow velocity (APV) in the PD was measured by transthoracic Doppler echocardiography at the time of intracoronary Doppler study. CFVR was defined as the ratio of hyperemic to basal APV. Clear envelopes of basal and hyperemic APV in the PD were obtained in 23 of 32 patients by transthoracic Doppler echocardiography. APV obtained from transthoracic Doppler echocardiography correlated highly with that from the Doppler guidewire method (r=0.91). The mean difference between transthoracic Doppler echocardiography and the Doppler guidewire method was 0.1+/-2.9. There was an excellent correlation between transthoracic Doppler echocardiography and the Doppler guidewire method for the measurements of CFVR (r=0.84). The mean difference between transthoracic Doppler echocardiography and Doppler guidewire was -0.016+/-0.198. Noninvasive measurement of coronary flow velocity and CFVR in the PD using transthoracic Doppler echocardiography accurately reflects invasive measurement of coronary flow velocity and CFVR by the Doppler guidewire method in pediatric patients with various heart diseases. PMID- 15793626 TI - Transcatheter closure of coronary-to-pulmonary fistula by nonconventional coils in a patient with tetralogy of Fallot. AB - We report a case in which a coronary-to-pulmonary fistula was successfully occluded in a patient with tetralogy of Fallot by employing the cut floppy tips of coronary angioplasty guidewires using a new technique. PMID- 15793625 TI - Late aortic root dilatation in tetralogy of Fallot may be prevented by early repair in infancy. AB - The objectives of this study were to examine the relative contributions of development and hemodynamics in aortic root dilatation of tetralogy of Fallot, to assess the impact of systemic to pulmonary artery shunt on aortic annular size, and to seek any relationship between the timing of corrective surgery and subsequent aortic root size. We performed a retrospective analytical study at a tertiary referral center of M-mode and two-dimensionol aortic root measurements in children with tetralogy of Fallot prior to any surgical or palliative intervention, after insertion of a surgical shunt, and on intermediate and long term follow-up post-repair. The main outcome measures were aortic root diameter prior to correction, after palliative shunt insertion, and after definitive repair. The study found that the aortic root is enlarged in unrepaired tetralogy of Fallot irrespective of age, increased in the first 3-6 months after palliative shunt surgery, and persisted into adulthood in those repaired after 1 year of age. Age-corrected aortic root dimensions normalized by mid-childhood in those who were repaired in infancy. Preexisting aortic root dilatation normalizes by 7 years of age in patients with tetralogy of Fallot who have been repaired in infancy, whereas it persists into adulthood in the group repaired post-infancy. PMID- 15793627 TI - Infected pseudoaneurysm of a right modified Blalock-Taussig shunt. PMID- 15793631 TI - Perioperative topical nitrate preserves sphincter function in patients undergoing transanal stapled anastomosis. PMID- 15793629 TI - Postconditioning--A new link in nature's armor against myocardial ischemia reperfusion injury. AB - Reperfusion injury is a complex process involving several cell types (endothelial cells, neutrophils, and cardiomyocytes), soluble proinflammatory mediators, oxidants, ionic and metabolic dyshomeostasis, and cellular and molecular signals. These participants in the pathobiology of reperfusion injury are not mutually exclusive. Some of these events take place during the very early moments of reperfusion, while others, seemingly triggered in part by the early events, are activated within a later timeframe. Postconditioning is a series of brief mechanical interruptions of reperfusion following a specific prescribed algorithm applied at the very onset of reperfusion. This algorithm lasts only from 1 to 3 minutes depending on species. Although associated with re-occlusion of the coronary artery or re-imposition of hypoxia in cell culture, the reference to ischemia has been dropped. Postconditioning has been observed to reduce infarct size and apoptosis as the "end games" in myocardial therapeutics; salvage of infarct size was similar to that achieved by the gold standard of protection, ischemic preconditioning. The cardioprotection was also associated with a reduction in: endothelial cell activation and dysfunction, tissue superoxide anion generation, neutrophil activation and accumulation in reperfused myocardium, microvascular injury, tissue edema, intracellular and mitochondrial calcium accumulation. Postconditioning sets in motion triggers and signals that are functionally related to reduced cell death. Adenosine has been implicated in the cardioprotection of postconditioning, as has e-NOS, nitric oxide and guanylyl cyclase, opening of K(ATP) channels and closing of the mitochondrial permeability transition pore. Cardioprotection by postconditioning has also been associated with the activation of intracellular survival pathways such as ERK1/2 and PI3 kinase - Akt pathways. Other pathways have yet to be identified. Although many of the pathways involved in postconditioning have also been identified in ischemic preconditioning, some may not be involved in preconditioning (ERK1/2). The timing of action of these pathways and other mediators of protection in postconditioning differs from that of preconditioning. In contrast to preconditioning, which requires a foreknowledge of the ischemic event, postconditioning can be applied at the onset of reperfusion at the point of clinical service, i.e. angioplasty, cardiac surgery, transplantation. PMID- 15793632 TI - Probiotic therapy to prevent pouchitis onset. PMID- 15793633 TI - Sphincter defects are not associated with long-term incontinence following ileal pouch-anal anastomosis. AB - INTRODUCTION: Frequent loose stools test the integrity of sphincter function in patients undergoing ileal pouch-anal anastomosis. The authors hypothesized that women with anal sphincter defects were more likely to experience incontinence episodes than women with intact sphincter muscles following ileal pouch-anal anastomosis. METHODS: From 1996 to 1998, 42 women with a mean age of 42 (range, 22-63) years were prospectively evaluated by anorectal manometry and endoanal ultrasound before pouch surgery. Forty women underwent a stapled ileal pouch-anal anastomosis and two underwent a handsewn anastomosis. All patients considered themselves continent of stool before the procedure. A postoperative survey including the Cleveland Clinic Florida scale, Fecal Incontinence Severity Index, and Fecal Incontinence Quality of Life scale was sent to study participants. RESULTS: Nineteen women with an obstetrical history had significant sphincter defects associated with significant lower mean resting pressure, mean squeeze pressure, and shorter anal canal length (3 vs. 3.7 cm, P = 0.0007). Thirty-five women (83 percent) responded resulting in a mean follow-up of 62 (range, 49-72) months. Fourteen responders (mean age, 46 years) had sphincter defects but no significant difference was found in Cleveland Clinic Florida scale, Fecal Incontinence Severity Index, or Fecal Incontinence Quality of Life scale scores when compared with those without defects. CONCLUSION: Although almost all women reported episodes of seepage, marked sphincter defects associated with low anal pressures and shorter anal canal length did not affect anal function following pouch surgery. This study supports the findings that continent women with significant sphincter defects on ultrasound evaluation may be considered for restorative proctocolectomy. PMID- 15793634 TI - Genetic testing for germline mutations of the APC gene in patients with apparently sporadic desmoid tumors but a family history of colorectal carcinoma. AB - PURPOSE: Desmoid tumors, also known as aggressive fibromatosis, occur with an incidence of 10 to 15 percent in patients affected by familial adenomatous polyposis, an autosomal inherited disease caused by germline mutations in the APC gene. However, sporadic forms with no hereditary background exist. The aim of this study was to find out whether there are APC germline mutations in apparently sporadic desmoid tumor patients without clinical or familial signs of familial adenomatous polyposis but with a family history of colorectal carcinoma in at least one family member. METHODS: Genomic DNA and mRNA were isolated from peripheral blood leukocytes of index patients of eight nonrelated families. Mutation screening was performed using reverse transcriptase polymerase chain reaction-based protein truncation test for APC exons 1-14. The large APC exon 15 was scrutinized by the protein truncation test of four overlapping genomic fragments. Additionally, genomic DNA from five desmoid tumors was analyzed for loss of heterozygosity at D5S346 close to the APC locus. RESULTS: No translational stop mutations typical for familial adenomatous polyposis could be found in the APC gene in any of the analyzed blood samples from the desmoid tumor patients. Additionally, no loss of heterozygosity at D5S346 was found in four of five desmoids; one tumor was not informative. CONCLUSIONS: These results may suggest that patients with sporadic desmoids and no clinical signs of familial adenomatous polyposis detected on careful examination, esophagogastroduodenoscopy, and complete colonoscopy do not need to be tested routinely for germline mutations of the APC gene. However, as large studies dealing with this problem are absent, it might be more time and cost effective to perform an APC mutational analysis instead. PMID- 15793635 TI - Recurrence rates after abdominal surgery for complete rectal prolapse: a multicenter pooled analysis of 643 individual patient data. AB - PURPOSE: This study was designed to determine what impact surgical technique, means of access, and method of rectopexy have on recurrence rates following abdominal surgery for full-thickness rectal prolapse. METHODS: Consecutive individual patient data on age, gender, surgical technique (mobilization-only, mobilization-resection-pexy, or mobilization-pexy), means of access (open or laparoscopic), rectopexy method (suture or mesh), follow-up length, and recurrences were collected from 15 centers performing abdominal surgery for full thickness rectal prolapse between 1979 and 2001. Recurrence was defined as the presence of full-thickness rectal prolapse after abdominal surgery. Chi-squared test and Cox proportional hazards regression analysis were used to assess statistical heterogeneity. Recurrence-free curves were generated and compared using the Kaplan-Meier method and log-rank test, respectively. RESULTS: Abdominal surgery consisted of mobilization-only (n = 46), mobilization-resection-pexy (n = 130), or mobilization-pexy (n = 467). There were 643 patients. After excluding center 8, there was homogeneity on recurrence rates among the centers with recurrences (n = 8) for age (hazards ratio, 0.6; 95 percent confidence interval, 0.2-1.7; P = 0.405), gender (hazards ratio, 0.6; 95 percent confidence interval, 0.1-2.3; P = 0.519), and center (hazards ratio, 0.3; 95 percent confidence interval, 0.1-1.5; P = 0.142). However, there was heterogeneity between centers with (n = 8) and without recurrences (n = 6) for gender (P = 0.0003), surgical technique (P < 0.0001), means of access (P = 0.01), and rectopexy method (P < 0.0001). The median length of follow-up of individual centers varied from 4 to 127 months (P < 0.0001). There were 38 recurrences at a median follow-up of 43 (range, 1-235) months. The pooled one-, five-, and ten-year recurrence rates were 1.06, 6.61, and 28.9 percent, respectively. Age, gender, surgical technique, means of access, and rectopexy method had no impact on recurrence rates. CONCLUSIONS: Although this study is likely underpowered, the impact of mobilization-only on recurrence rates was similar to that of other surgical techniques. PMID- 15793637 TI - Laparotomy for acute colorectal conditions in moribund patients: is it worthwhile? AB - INTRODUCTION: Colorectal surgeons are sometimes asked to operate on moribund patients, classified by the American Society of Anesthesiology as Class 5- patients who are likely to die with or without surgery. There is a concern that surgery may precipitate the patient's demise; however, there are few data concerning outcomes in this group. It is, therefore, difficult to counsel patients and their families about the advisability of surgery. This study was performed to show what sort of results can be expected from operating on American Society of Anesthesiology Class 5 patients and to see whether there are any factors that can be used to predict outcome within this American Society of Anesthesiology class. METHODS: Since March 1989, a prospective database of all surgeries has been maintained. Included in the data is the American Society of Anesthesiology Classification. All patients classified as 5 were identified and their history reviewed. The 30-day postoperative survival was the primary end point of the study. Also abstracted were data on comorbidity, indication for colorectal surgery, surgery performed, length of postoperative hospital stay, estimated intraoperative blood loss, and duration of anesthesia. RESULTS: There were 4,163 surgeries recorded, including 2,040 (49 percent) laparotomies. There were 21 laparotomies in American Society of Anesthesiology Class 5 patients (1 percent of all laparotomies). Eighteen surgeries were performed as emergencies, and three were urgent. Nine patients survived 30 days or longer (41 percent), whereas 12 did not. Seven patients lived to be discharged from the hospital. CONCLUSION: Expeditious, safe surgery may save moribund patients with an acute colorectal problem. PMID- 15793636 TI - Aberrant methylation of HLTF, SOCS-1, and CDH13 genes is shown in colorectal cancers without lymph node metastasis. AB - PURPOSE AND METHODS: In this study, we examined the combined methylation status of HLTF, SOCS-1, and CDH13 in 61 resected primary colorectal cancers using methylation-specific polymerase chain reaction and correlated the number of methylated genes with the clinicopathologic features of affected patients. RESULTS: We found a significant difference in lymph node metastasis (P = 0.020) when we compared the number of methylated genes in colorectal cancers with lymph node metastasis to those without it. CONCLUSIONS: Colorectal cancers without lymph node metastasis frequently exhibited the aberrant methylation of HLTF , SOCS-1 , and CDH13 genes. PMID- 15793638 TI - Is laparoscopic colectomy applicable to patients with body mass index >30? A case matched comparative study with open colectomy. AB - PURPOSE: The benefits of early postoperative recovery, reduced postoperative pain, pulmonary dysfunction, and hospitalization after laparoscopic colectomy may improve outcome over open colectomy in obese patients. This case-matched study compares outcomes after open and laparoscopic colectomy. METHODS: A total of 94 laparoscopic colectomy patients with a body mass index >30 (Jan 1999-June 2003) were identified from a prospective database and matched to open colectomy cases for age, gender, body mass index, American Society of Anesthesiologists class, procedure, indication, and date of surgery. Operating time, length of stay, conversion, intraoperative and postoperative complications, reoperation, 30-day readmission rate, and costs were compared. Data are presented as means +/- standard deviations, and appropriate statistical tests were used. RESULTS: The two groups were matched for age (P = 0.06), gender (P = 1), American Society of Anesthesiologists class (P = 0.2), body mass index (P = 0.4), indication for surgery (P = 1), and procedure (P = 1). By using intention-to-treat-type analysis, there was no difference in median operating time (100 vs. 110 (mean, 123 vs. 112) minutes; P = 0.1), complications (21 vs. 24 percent; P = 0.74), readmission (17 vs. 10.6 percent; P = 0.3), reoperation rates (6.4 vs. 4.3 percent; P = 0.75), or direct costs (median, US. 3,368 dollars vs. US 3,552 dollars; mean, US 4,003 dollars vs. US 4,037 dollars; P = 0.14) between laparoscopic colectomy or open colectomy; however, the median length of stay (3 vs. 5.5 (mean, 3.8 vs. 5.8) days; P = 0.0001) was significantly shorter after laparoscopic colectomy. Twenty-eight patients required conversion for adhesions (n = 11), bleeding (n = 3), obesity-hindering vision or dissection (n = 9), large phlegmon or tumor (n = 4), and ureteric injury (n = 1). The mean operating time for conversions was 142 minutes and length of stay was 6.4 days. Compared with laparoscopically completed cases, the median length of stay (5 vs. 2 (mean, 6.4 vs. 2.8) days; P = 0.0001) and median operating times (150 vs. 95 (mean, 142 vs. 115) minutes; P = 0.02) were significantly higher in the converted group, but there was no difference in the complication (P = 0.8), readmission (P = 1), or reoperation (P = 0.7) rates. Compared with open colectomy, the operating time (P = 0.02) was significantly higher in the converted group but there were no significant differences in the length of stay (P = 0.18), complication (P = 1), readmission (P = 0.35), or reoperative (P = 1) rates. CONCLUSIONS: Laparoscopic colectomy can be performed safely in obese patients, with shorter postoperative recovery than that with open colectomy. Although obesity is associated with a high conversion rate, outcome in these converted cases is comparable to the matched open cases. PMID- 15793639 TI - Prospective evaluation of factors predicting difficulty and pain during sedation free colonoscopy. AB - PURPOSE: Colonoscopy can be painful for patients and difficult for colonoscopists; however, it is hard to predict how painful or difficult the examination will be. This study was designed to identify factors that predict pain and difficulty during sedation-free colonoscopy. METHODS: A total of 848 consecutive sedation-free colonoscopies were evaluated in a prospective manner. Factors were recorded, including patient pain, intubation time, demographic data, history of abdominal surgery, bowel preparation status, diverticular disease, bowel habits, anxiety level, and number of previous colonoscopies. These factors were analyzed to determine their association with pain and difficulty. RESULTS: Almost all colonoscopies (845/848; 99.6 percent) were successful. Univariate analyses showed that lower body mass index, younger age, female gender, anxiety level, first time, intubation time, preparation status, previous hysterectomy, and previous gynecologic surgery were predictors of patient pain, and lower body mass index, female gender, anxiety level, preparation status, previous hysterectomy, previous gynecologic surgery, and constipation were predictors of difficulty of intubation. Multivariate logistic regression analyses revealed that lower body mass index, younger age, intubation time, preparation status, previous hysterectomy, and antispasmodic agent use were predictors of patient pain, and lower body mass index, female gender, constipation, preparation status, and previous hysterectomy were predictors of difficulty of intubation. CONCLUSIONS: By use of intubation time and patient pain, several patient characteristics were identified that may predict technical difficulty and pain associated with the procedure. These findings have implications for the practice and teaching of colonoscopy. PMID- 15793640 TI - Intersphincteric resection in very low rectal cancer. PMID- 15793641 TI - Curative resection of T1 colorectal carcinoma: risk of lymph node metastasis and long-term prognosis. AB - PURPOSE: The features of T1 colorectal adenocarcinoma and the risk determination of lymph node metastasis were reviewed. Prognostic factors were assessed to verify whether the risk of lymph node metastasis would influence the long-term prognosis. METHODS: Patients undergoing curative resection of T1 colorectal adenocarcinoma at the Taipei Veterans General Hospital from December 1969 to August 2002 were retrospectively studied. Patients with synchronous colorectal cancer, distant metastasis, familiar adenomatous polyposis, or inflammatory bowel disease were excluded. The associations between lymph node metastasis and clinicopathologic variables were evaluated univariately using chi-squared test, Fisher's exact test, or Student's t -test, and multivariately using logistic regression. Univariate analysis by the log-rank test and multivariate analysis by Cox regression hazards model determined the factors influencing the overall survival. RESULTS: A total of 159 patients were included. Sixteen patients (10.1 percent) had lymph node metastasis. The risk of lymph node metastasis included histologic grade (P = 0.005), lymphatic vessel invasion (P = 0.023), inflammation around cancer (P = 0.049), and budding at the invasive front of tumor (P = 0.022). Age (P = 0.001) and number of total sampling lymph nodes (P < 0.0001) were found to be the factors influencing the overall survival. CONCLUSIONS: Variables that predict lymph node metastasis in surgically resected T1 colorectal carcinoma may not impact the long-term prognosis. PMID- 15793642 TI - Management of anal canal cancer. AB - PURPOSE: Chemoradiotherapy has replaced radical surgery as the initial treatment of choice for anal canal cancer. The roles of these therapeutic modalities are discussed and recommendations on management of anal canal cancer are made based on currently available evidence. Areas for further studies also are identified. METHODS: Literature on management of anal canal cancer from January 1970 to July 2003 obtained via MEDLINE was reviewed. Reports on anal margin cancers were excluded. RESULTS: Randomized, prospective, Phase 3 trials in Europe and the United States showed that chemoradiotherapy with 5-fluorouracil and mitomycin C was superior in local control, colostomy-free rate, progression-free survival, and cancer-specific survival compared with radiation alone. In larger tumors, the addition of mitomycin C to radiotherapy and 5-fluorouracil improves local control, colostomy-free, and disease-free survival but is associated with more acute hematologic toxicity. Chemoradiotherapy, including Cisplatin and 5 fluorouracil, appeared to be equal or superior to surgery as salvage therapy in patients with residual disease six weeks after initial nonsurgical treatment. CONCLUSIONS: To improve treatment outcomes and reduce treatment-related toxicities, further studies are required to elucidate the optimal drug combination and doses, optimal radiation field, total dose, and fraction sizes. Randomized, multicenter trials are needed to define the treatment protocol that provides the highest rate of sphincter preservation with acceptable toxicity. Few studies addressed the treatment of metastatic disease, which remains a major cause of mortality. PMID- 15793643 TI - Magnetic resonance imaging of the rectum during distension. AB - PURPOSE: A knowledge of the relationships between the rectum and its surrounding structures during distention may improve our understanding of the results of studies assessing rectal sensory-motor responses to distention. This magnetic resonance imaging study was designed to assess the shape of the rectum and the degree of distention at which the surrounding structures are compressed. METHODS: Nine healthy patients underwent magnetic resonance imaging of the rectum under resting conditions and after the inflation of a plastic bag to volumes of 50, 100, 150, 200, and 250 ml. The thickness of the rectovesical space was assessed as a measure of the compression of the perirectal structures, and the perception of sensations were recorded. RESULTS: The shape of the rectum changed from being quasicylindrical at distention volumes of <100 ml to bean-shaped at larger volumes. The thickness of the rectovesical space at a distention volume of 50 ml was the same as when the bag was not inflated, but it progressively decreased until the difference became statistically significant at distention volumes of > or = 200 ml, corresponding to a mean +/- standard deviation rectal radius of 2.66 +/- 0.37 cm. Statistically significant compression of the rectovesical space was recorded when the sensations of gas, desire to defecate, and urgency were perceived. CONCLUSIONS: The shape of the rectum changes during distention; it significantly compresses the extrarectal structures in the tested range of distention that induces non-painful sensations. Magnetic resonance imaging is a useful means of assessing the morphologic changes in the rectum during distention. PMID- 15793644 TI - Does hemodynamic instability predict positive technetium-labeled red blood cell scintigraphy in patients with acute lower gastrointestinal bleeding? A review of 50 patients. AB - PURPOSE: Technetium-99m-labeled red blood cell scintigraphy, commonly used in the evaluation of acute lower gastrointestinal hemorrhage, often fails to demonstrate a source of bleeding. It would be helpful to characterize a subset of patients more likely to have a positive scan. This study was undertaken to determine whether hemodynamic instability can predict tagged red blood cell scan positivity. METHODS: The records of 50 consecutive patients who underwent tagged red blood cell scanning for the evaluation of acute lower gastrointestinal bleeding were reviewed retrospectively. RESULTS: Patients presenting with a heart rate >100 beats per minute or a systolic blood pressure <100 mmHg up to 24 hours before undergoing tagged red blood cell scanning were considered hemodynamically unstable. Thirteen of 21 unstable patients (62 percent) had positive scans, whereas only 6 of 29 stable patients (21 percent) had positive scintigraphy (odds ratio, 6; 95 percent confidence interval, 1.79-22.1). CONCLUSIONS: Hemodynamic instability in the setting of acute lower gastrointestinal bleeding may be a predictor of positive tagged red blood cell scanning. Incorporating this into the diagnostic algorithm used to evaluate patients with acute lower gastrointestinal bleeding may allow physicians to reserve red blood cell scintigraphy for patients who have demonstrated transient hemodynamic compromise. PMID- 15793645 TI - Surgical salvage of recurrent rectal cancer after transanal excision. AB - PURPOSE: This study examines surgical salvage of locally recurrent rectal cancer following transanal excision of early tumors. METHODS: Through retrospective review of a colorectal database we identified 50 patients who underwent attempted surgical salvage for local recurrence following initial transanal excision of T1 or T2 rectal cancer. Eight patients had resectable synchronous distant disease. Clinicopathologic variables were associated with extent of surgery required for salvage and outcome. RESULTS: Salvage procedures included abdominoperineal resection (31), low anterior resection (11), total pelvic exenteration (4), and transanal excision (3). One patient had unresectable disease at exploration, requiring diverting ostomy. Of the 49 patients who underwent successful salvage, 27 (55 percent) required an extended pelvic dissection with en bloc resection of one or more of the following structures: pelvic sidewall and autonomic nerves (18); coccyx or portion of sacrum (6); prostate (5); seminal vesicle (5); bladder (4); portion of the vagina (3); ureter (2); ovary (1); and uterus (1). Complete pathologic resection (R0) was accomplished in 47 of 49 patients. Of the eight patients with distant and local recurrence, two underwent synchronous resection and six had delayed metastasectomy. With a median follow-up of 33 months, 29 patients had recurred or died of disease at the time of this analysis. Five-year disease-specific survival was 53 percent. Factors predictive of survival included evidence of any mucosal recurrence on endoscopy, low presalvage carcinoembryonic antigen, and absence of poor pathologic features (lymphovascular and perineural invasion). Patients who required an extended pelvic resection had a worse survival rate. CONCLUSION: Pelvic recurrence following transanal excision of early rectal cancer is often locally advanced, requiring an extended pelvic dissection with en bloc resection of adjacent pelvic organs to achieve salvage. The long-term outcome in patients undergoing resection is less than expected, considering the early stage of their initial disease. When contemplating local excision for early rectal cancer, the risk of local recurrence, the extent and morbidity of surgery required for salvage, and the modest cure rate following salvage should be considered. PMID- 15793646 TI - Neoadjuvant imatinib in gastrointestinal stromal tumor of the rectum: report of a case. AB - Gastrointestinal stromal tumors are rare tumors of the gastrointestinal tract. Gastrointestinal stromal tumors involving the rectum are uncommon. We describe a case of a 43-year-old female with a gastrointestinal stromal tumor of the rectum who declined abdominoperineal resection. Neoadjuvant treatment with imatinib decreased her tumor size, permitting sphincter-sparing transanal excision. She had no evidence of disease for 24 months postoperatively until she recurred with lung metastases. Microdissection genotyping of the recurrent lesion revealed a deletion in exon 11. Further mutational analysis showed that her metastatic lesion was concordant with her primary rectal lesion, suggesting that systemic micrometastasis was previously present at initial diagnosis. Deletion in exon 11 predicts for response with imatinib treatment and is associated with a longer event-free and overall survival. Current studies are underway that may help us optimize the treatment for patients with gastrointestinal stromal tumors. PMID- 15793647 TI - Sacral nerve stimulation for fecal incontinence following surgery for rectal prolapse repair: a multicenter study. AB - PURPOSE: A proportion of patients have fecal incontinence secondary to a full thickness rectal prolapse that fails to resolve following prolapse repair. This multicenter, prospective study assessed the use of sacral nerve stimulation for this indication. METHODS: Patients had to have more than or equal to four days with fecal incontinence per 21-day period more than one year after surgery. They had to have failed conservative treatment and have an intact external anal sphincter. RESULTS: Four female patients aged 42, 54, 68, and 65 years met the inclusion criteria. Three of the four patients had had more than one operation for recurrent full-thickness rectal prolapse before sacral nerve stimulation, one of whom had undergone a further operation for recurrence following stimulation. One patient had undergone one operation for prolapse repair. The preoperative duration of symptoms was ten, eight, three, and nine years, respectively. Although patients had an intact external anal sphincter, one patient had a fragmented internal anal sphincter. The frequency of fecal incontinent episodes changed from 11, 24.7, 5, and 8 per week at baseline to 0, 1.5, 5.5, and 1 per week at latest follow-up. Ability to defer defecation was also improved in two of three patients who had this documented. Fecal incontinence-specific quality of life assessment showed an improvement in all four domains. CONCLUSION: Sacral nerve stimulation should be considered for patients with ongoing fecal incontinence following full-thickness rectal prolapse repair if they prove resistant to conservative treatment. PMID- 15793648 TI - Stapled hemorrhoidopexy vs. Harmonic Scalpel hemorrhoidectomy: a randomized trial. AB - PURPOSE: A randomized trial was undertaken to evaluate and compare stapled hemorrhoidopexy with excisional hemorrhoidectomy in which the Harmonic Scalpel was used. METHODS: Patients with Grade III hemorrhoids who were employed during the trial period were recruited and randomized into two groups: (1) Harmonic Scalpel hemorrhoidectomy, and (2) stapled hemorrhoidopexy. All operations were performed by a single surgeon. In the stapled group, the doughnut obtained was sent for histopathologic examination to determine whether smooth muscles were included in the specimen. Operative data and complications were recorded, and patients were followed up through a structured pro forma protocol. An independent assessor was assigned to obtain postoperative pain scores and satisfaction scores at six-month follow-up. Patients were also administered a simple questionnaire at follow-up to assess continence functions. RESULTS: Over a 20-month period, 88 patients were recruited. The two groups were matched for age and gender distribution. No significant difference was identified between the two groups in terms of operation time, blood loss, day of first bowel movement after surgery, and complication rates. Despite a similar parenteral and oral analgesic requirement, the stapled group had a significantly better pain score (P = 0.002); these patients also had a significantly shorter length of stay (P = 0.02), and on average resumed work nine days earlier than the group treated with the Harmonic Scalpel (6.7 vs. 15.6, P = 0.002). Although 88 percent of doughnuts obtained in the stapled group contained some smooth muscle fibers, no association was found between smooth muscle incorporation and postoperative continence function, and as a whole the continence outcomes of the stapled group were similar to those after Harmonic Scalpel hemorrhoidectomy. Finally, at six-month follow-up, patients who underwent the stapled procedure had significantly better satisfaction scores (P = 0.001). CONCLUSION: Stapled hemorrhoidopexy is a safe and effective procedure for Grade III hemorrhoidal disease. Patients derive greater short-term benefits of reduced pain, shorter length of stay, and earlier resumption to work. Long-term follow-up is necessary to determine whether these initial results are lasting. PMID- 15793649 TI - Manometric effect of topical glyceryl trinitrate and its impact on chronic anal fissure healing. AB - INTRODUCTION: The duration of physiologic action of topical glyceryl trinitrate in the management of anal fissure has been the source of some controversy. This study was designed to assess the manometric effect of glyceryl trinitrate on internal sphincter resting tone with continuous monitoring. METHODS: Twenty-seven patients with a chronic anal fissure were assessed with fissure, pain, bleeding, and continence scores. Twenty-two were randomized to 1 cm of topical 0.2 percent glyceryl trinitrate paste, applied to the lower anal canal. Five patients were randomized to 1 cm of water-soluble lubricating jelly to the lower anal canal. Continuous stationary six radial channel water perfusion anorectal manometry was performed for 5 minutes before treatment and then for a further 30 minutes. The 22 glyceryl trinitrate patients were then advised to apply topical 0.2 percent glyceryl trinitrate, three times daily, for eight weeks. Twenty-four hours after completing treatment, all baseline assessments were repeated. The lubricant jelly cohort was discharged from the study after the initial assessment. RESULTS: During the initial manometric assessment, 21 glyceryl trinitrate patients (95 percent) had 20 percent or more reduction in mean and maximum anal resting pressure after treatment. However, there was no statistical difference at 20 minutes compared with 0 minutes (P > 0.1). After eight-week treatment, 16 patients (73 percent) reported symptom resolution and 15 (67 percent) were found to be healed on examination. Clinical healing and resolution of symptoms positively correlated with a higher pretreatment maximum anal resting pressure in the mid anal canal (P < 0.0001), lower fissure score (P < 0.0001), and greater percentage reduction of the maximum resting pressure after application of glyceryl trinitrate (P < 0.001). The mean and maximum anal resting pressure at Week 8 was not significantly different from the baseline values (P > 0.05). During continuous manometry, the anal resting pressure did not significantly change in the patients treated with lubricating jelly. CONCLUSIONS: In those patients with a lower fissure score, a higher mid anal canal anal resting pressure, and a greater resting pressure reduction after glyceryl trinitrate application, a favorable clinical outcome can be expected with glyceryl trinitrate treatment. However, because the physiologic response has resolved in fewer than 20 minutes, the dosing regime should be reassessed. PMID- 15793651 TI - Profound hypothermia with alpha-stat pH management during open-heart surgery is associated with choreoathetosis. AB - A retrospective chart review was conducted to examine risk factors associated with the occurrence of choreoathetosis, a rare but significant complication of open-heart surgery in children. Ten children were identified as having developed choreoathetosis after cardiac surgery. Their charts were reviewed and compared with 33 age- and diagnosis-matched controls who underwent open-heart surgery during the same time period. Children with choreoathetosis reached lower rectal and esophageal temperatures (p = 0.0018 for both) and spent a greater portion of total bypass time at lower rectal and esophageal temperatures (p < 0.001 for both). Duration of cooling below 20 degrees C esophageal temperature and PaCO2 at the end of the cooling period were significant predictors of choreoathetosis (p = 0.023 and p = 0.0497, respectively) in a logistic regression model, and a greater fraction of choreoathetosis patients had prior developmental delays (p = 0.017). No difference was found in the age at surgery, duration of bypass, aortic cross clamp time, arterial pH, PaCO2 or mean arterial pressure. The combination of extended exposure to profound hypothermia and alpha-stat pH management strategy and preexisting developmental delay are associated with the development of choreoathetosis following open-heart surgery 61 in children. PMID- 15793652 TI - Effect of age on diurnal changes of 24-hour QT interval variability. AB - QT interval on the surface electrocardiogram (ECG) reflects the time for repolarization of myocardium and prolongation of QTc is strongly associated with sudden cardiac death. Studies using novel techniques on beat-to-beat QT interval variability (QTV) have shown that it is influenced by the autonomic nervous system and is a predictor of sudden cardiac death. In this study, we examined the awake and sleep changes in QTV in 39 normal adults (mean age, 35 years) and 10 children (mean age, 11 years) using 24-hour ECG records. We obtained eight 5 minute segments of ECG sampled at 1000 Hz from the 24-hour records. Our results show that there is a diurnal variation of QTvm, detrended QT interval variance corrected for mean QT, and QTvi, an index of QTvm divided by heart rate variability corrected for mean heart rate. There was a significant increase in mean QT during sleep, whereas there was a significant decrease in QTvm and QTvi. QTvi significantly increased during the early morning hours. There were significant but modest correlations between the average 24-hour and awake QTvi and age (p < 0.01). There were also decreased low-frequency and high-frequency powers of QT during sleep. Coherence between heart rate and QT interval fluctuations in the range of 0-0.5 Hz, and especially in the high-frequency range (0.15-0.5 Hz), was significantly lower in adults than in children (p < 0.0002). These findings demonstrate diurnal fluctuations in ventricular repolarization lability. We speculate that these effects may relate to changes in cardiac autonomic function and may contribute to the well-known diurnal variation in the incidence of ventricular arrhythmias. PMID- 15793653 TI - Cardiac troponin T: a marker in the diagnosis of acute myocarditis in children. AB - This study was conducted to assess the use of serum cardiac troponin T (cTnT) level as a noninvasive indicator to diagnose acute myocarditis in children. Noninvasive conventional methods often fail to diagnose myocarditis, A median cTnT level of 0.088 ng/ml (0.04-3.11) was reported in pediatric patients with acute myocarditis in our previous study. Hence, we attempted to determine the cutfoff level of cTnT to diagnose acute myocarditis in children. Pediatric patients with clinically suspected myocarditis or dilated cardiomyopathy (DCM) and a control group were recruited. History, physical examination, elctrocardiogram, chest roentgenogram, echocardiogram, cTnT level, and/or endomyocardial biopsy and clinical course were studied. The gold standard to diagnose acute myocarditis was endomyocardial biopsy proved according to the Dallas criteria and/or recovery from cardiovascular problems within 6 months of follow-up. Forty-three patients were admitted due to cardiovascular problems from primary myocardial dysfunction. Twenty-four patients were diagnosed as acute myocarditis (group 1), 19 were idiopathic chronic DCM (group 2), and 21 patients had moderate to large ventricular septal defect and congestive heart failure (group 3). Median cTnT level was statistically higher in (group 1) compared to groups 2 and 3. Ejection fraction (EF) and left ventricular end diastolic dimension (LVEDd) z score of acute myocarditis were 38.5% (range, 21-67) and 1.3 (range, -0.8-3.0), respectively, which were significantly better than DCM [28.0% (range, 17-45) and 6.0 (range, 2.0-10.0)]. The cutoff point of cTnT level to diagnose acute myocarditis was 0.052 ng/ml (sensitivity, 71%; specificity, 86%). cTnT level, EF, and LVEDd z score did not predict short-term outcomes of patients. In acute myocarditis, cTnT level and EF were significantly higher and LVEDd z score was significantly lower than in DCM. However, the three parameters had no significant effect on outcomes of the patients. Our data show that cardiac a cTnT level of 0.052 ng/ml is an appropriate cutoff point for the diagnosis of acute myocarditis. PMID- 15793654 TI - Patent foramen ovale morphology and impact on percutaneous device closure. AB - We sought to evaluate the effects of atrial septal and patent foramen ovale (PFO) morphology on the efficacy of transcatheter closure. We performed a retrospective analysis of all patients who underwent PFO device closure from January 1997 to January 2002. Forty-seven patients underwent percutaneous closure of a PFO with a median age of 45 years (range, 8-75) and weight of 76 kg (range 28-115). The septal morphology was flat in 33 and aneurysmal in 14 patients. The PFO morphology was a simple flap in 20 and complex in 27 patients. Complex morphologies included long-tunnel PFO (n = 15), coexistent small atrial septal defect (ASD) (n = 5), and aneurysmal septum without a tunnel or ASD (n = 7). Nonstretched PFO diameters were significantly smaller than stretched (4.8 +/- 1.1 mm vs 11.6 +/- 3.8 mm, p < 0.01). Median device size: stretched diameter ratio (DS:SD) was 3.7:1 (range, 2.2-9.1). The DS:SD ratio was significantly higher in patients with complex PFO (mean, 3.9:1 vs 2.6:1; p < 0.05). Device placement was successful in all patients. Five patients required transeptal puncture of the foraminal flap in long-tunnel PFOs. Effective closure on follow-up was achieved in 45 patients (95%). Of the 2 patients with residual shunts, 1 had a complex PFO and the other a simple PFO. Two patients (5%) experienced recurrent neurologic symptoms after device closure despite having no residual shunt by echocardiography. Complex PFO did not increase risk of residual shunt or recurrent neurologic symptoms after device closure. PMID- 15793655 TI - Echocardiographic and electrocardiographic findings of cardiomyopathy in Duchenne and Becker-Kiener muscular dystrophies. AB - We assessed the cardiac findings in Duchenne muscular dystrophy (DMD) and Becker Kiener muscular dystrophy (BMD) patients in the large outpatient group of our single-center institution. The analysis included the revision of patient records (retrospectively) and current echocardiography, electrocardiogram (ECG), and Holter ECG data (prospectively). Reduced left ventricular fraction shortening (FS) < 25% was found in 24% of all patients (50 DMD, 12.1 +/- 4.7 years: 20 BMD, 17.1 +/- 8.5 years). Median age of onset of FS < 25% was 16.8 +/- 1.0 in DMD and 30.4 +/- 3.4 in BMD; (p < 0.05). Holter ECG in DMD demonstrated sinus tachycardia in 26% deprivation of circadian rhythm in 31%, and reduced heart rate variability in 51%. For these reasons, we recommend carrying out echocardiography annually in DMD and BMD > 10 years. Because the rate of disturbances in the cardiac autonomic nervous system is high in DMD, we also recommend Holter ECG annually. Further investigation should be conducted to determine if treatment with ss-blockers and ACE inhibitors in combination reduces cardiac autonomic nervous imbalance, thus improving patient outcome in DMD. PMID- 15793657 TI - Use of tissue plasminogen activator for femoral artery thrombosis following transcatheter coil occlusion of patent ductus arteriosus. AB - Femoral artery thrombosis is an uncommon but potentially serious complication following pediatric cardiac catheterization. Management options include heparin infusion, thrombolytic therapy, and surgical thrombectomy. The use of thrombolytic agents following coil occlusion of shunts, collaterals, and patent ductus arteriosus (PDA) may be successful in resolving the femoral arterial thrombosis but may also reopen the device-occluded vessel. We report the successful use of tissue plasminogen activator for management of femoral artery thrombosis in a child following transcatheter PDA coil occlusion in which the PDA remained occluded. PMID- 15793656 TI - High association of congenital heart disease with indirect inguinal hernia. AB - We evaluated 103 patients with indirect inguinal hernia (IIH) for the association of congenital heart disease by echocardiography. Congenital cardiac abnormalities were recognized in 32% of patients with inguinal hernia, which is significantly higher than that reported in a population-based study in Turkey and other population-based studies. Ventricular septal defect and valvular anomalies are the most frequently detected malformations. Our findings suggest that screening for congenital hearth disease is necessary in children with indirect inguinal hernia. PMID- 15793659 TI - Advantages of oxygen inhalation therapy for postoperative pulmonary hypertension. AB - Total correction was performed in a case of complete transposition of the great arteries (TGA) with severe pulmonary vascular obstructive disease (PVOD). Although severe pulmonary hypertension remained after surgery, oxygenation was continued for 15 months, which included a shift to at-home oxygen inhalation therapy (HOT). Cardiac catheterization 15 months after surgery demonstrated that pulmonary hypertension was greatly improved. For patients in whom the palliative Mustard operation is considered due to severe PVOD on the basis of lung biopsy diagnosis, total correction of TGA is possible by employing HOT after surgery. PMID- 15793658 TI - Improvement of left ventricular function after changing the pacing site in a child with isolated congenital complete atrioventricular block and dilated cardiomyopathy. AB - We report a case of isolated congenital complete atrioventricular block with left ventricular dysfunction after pacemaker implantation that improved after the pacing site was changed. During the neonatal period, a pacemaker wire was implanted on the right ventricular epicardium and pacing was initiated. Decreased ejection fraction and a perfusion defect around the septum on myocardial scintigraphy were observed during follow-up. Induced left bundle branch block was thought to be causing interventricular asynchrony, and the pacing site was change to the left ventricular epicardium. Ejection fraction improved and the perfusion defect resolved. Lead relocation may be useful for left ventricular dysfunction that develops during right ventricular pacing. PMID- 15793660 TI - Fenestration in extracardiac conduits in children after modified Fontan operation by implantation of stent grafts. AB - Three patients (ages 3, 5, and 8 years) with various forms of functionally univentricular heart lesions received a total cavopulmonary connection with an extracardiac conduit as a final reconstructive procedure. Failure of the Fontan circulation occurred immediately after surgery because of spontaneous closure of surgical fenestrations in two children and absent fenestration in one. As an emergency procedure, in all patients the conduit was perforated by transcatheter intervention in order to create a connection to the anatomical right atrium. Following balloon dilatation of the perforated conduit, in all three patients covered stent grafts were placed in the newly created defect to attain a reliable communication. Patency of the fenestration was demonstrated by angiogram and any leakage was ruled out. Cardiac output improved and severe pleural effusion and ascites subsided. Right-to-left shunt could be demonstrated by echocardiography at follow-up after 7 months (median) in all three patients. Oxygen saturation remained stable between 85 and 90%. These preliminary results suggest that stent graft fenestration can serve as a valuable tool in failing Fontan circulation, particularly in patients with an extracardiac conduit. Covered stents have the potential to reduce the acute risk of bleeding and they help to prevent early spontaneous closure of the newly created fenestration. PMID- 15793663 TI - Time intervals (3' or 5') between dose steps can influence methacholine challenge test. AB - Bronchial hyperresponsiveness (BHR) is a common feature in the majority of asthmatic subjects and methacholine is the most frequently used agent for the test. The influence of 3 or 5 min time intervals between doses steps in a double methacholine challenge test (MCH-3' or MCH-5') was investigated. Using the MCH-3' challenge, 52 intermittent asthmatics were classified as having moderate (BHR-M; 18 subjects), mild (BHR-m; 19 subjects), or bordeline (BHR-B; 15 subjects) BHR. The cumulative dose and the PD20FEV(1) were higher for MCH-5' compared with MCH 3' in BHR-m (p < 0.05) and BHR-B (p < 0.05) but not in the BHR-M group. Also the dose response slopes, FEV(1)% decline/cumulative methacholine dose, calculated for the two challenge tests were statistically different only in BHR-m (p < 0.05) and BHR-B (p < 0.01). At MCH-5', there were 16 subjects with BHR-M, 18 with BHR m, 12 with BHR-B and 6 subjects with normal reactivity. Results may suggest that in the group of BHR-m and BHR-B subjects, at MCH-5' compared with MCH-3', the cumulative effect of the administered drug, quickly metabolized by cholinesterase, is not complete, thus leading to an incorrect estimation of bronchial hyperresponsiveness degree. It is hoped that time interval between doses be standardized to ensure maximum comparability within and between subjects in challenge tests. PMID- 15793661 TI - Post-hoc diagnosis of congenital long QT syndrome in patients with tetralogy of Fallot. AB - This report describes two patients diagnosed with congenital long QT syndrome after surgical repair of tetralogy of Fallot. Despite the fact that both patients had preoperative electrocardiograms demonstrating QT prolongation, neither was diagnosed until long after their surgeries, when they or their relatives presented with symptoms of long QT syndrome. A brief discussion highlights the reasons why long QT syndrome may be overlooked in patients with structural heart defects and the clinical importance of identifying these patients preoperatively. PMID- 15793664 TI - The role of pleural fluid-serum gradient of tumor necrosis factor-alpha concentration in discrimination between complicated and uncomplicated parapneumonic effusion. AB - In a previous preliminary study an excess of tumor necrosis factor-alpha (TNF) was found in pleural fluid of patients with complicated parapneumonic effusion (CPPE), and its levels in pleural fluid of these patients were shown to be significantly higher than those in patients with uncomplicated parapneumonic effusion (UCPPE). This larger population study was undertaken to investigate, for the first time, the role of pleural fluid-serum gradient of TNF (TNFgradient) in discrimination between UCPPE and CPPE. Using a commercially available high sensitivity ELISA kit, levels of TNF were measured in serum and pleural fluid of 51 patients with UCPPE and 30 patients with nonempyemic CPPE. The mean +/- SEM values of serum TNF (TNFserum), pleural fluid TNF (TNFpf), and TNFgradient in the UCPPE group were 6.65 +/- 0.48 pg/mL, 10.85 +/- 0.74 pg/mL, and 4.2 +/- 0.38 pg/mL respectively, and in the CPPE group they were 7.59 +/- 0.87 pg/mL, 54.02 +/ 5.43 pg/mL, and 46.43 +/- 5.34 pg/mL, respectively. While no significant difference was found between the two groups regarding levels of TNFserum (p = 0.31), a highly significant difference between these two groups was found regarding levels of TNFpf and TNFgradient (p < 0.0001 for both variables). A significant correlation was found between levels of TNFserum and levels of TNFpf in the UCPPE group (r = 0.89, p < 0.0001), but not in the CPPE group (r = 0.18, p < 0.33). TNFgradient at an optimal cut-off level of 9.0 pg/mL was found to be a good marker for discrimination between UCPPE and CPPE (sensitivity, 96.7%, specificity, 98%, accuracy, 97.5%, and p < 0.0001). In conclusion, levels of TNFpf but not TNFserum are significantly higher in CPPEs than those in UCPPEs where TNFgradient at an optimal cut-off level of 9.0 pg/mL is a good marker for discrimination between UCPPE and CPPE. PMID- 15793665 TI - Role of serum vascular endothelial growth factor in the prediction of angiogenesis and prognosis for non-small cell lung cancer. AB - Although vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP)-9 are involved in angiogenesis of various cancers, clinical utility of preoperative serum concentration of these molecules in non-small cell lung cancer (NSCLC) has not yet been elucidated. In this study, we determined the concentration of VEGF, MMP-9 and various other tumor markers in serum prior to surgery and evaluated the results compared with intratumoral vasculature to isolate a valuable marker in determining the prediction of angiogenesis in NSCLC. Among these molecules and serum tumor markers, circulating serum VEGF was identified to markedly correlate with microvessel density (MVD) of the resected tumor specimens. Moreover, overall survival of patients with low VEGF levels (< or =326 ng/ml) was significantly greater than that of patients with high VEGF levels (>326 ng/ml), while patients with low MMP-9 levels (< or =189 ng/ml) and those with high MMP-9 levels (>189 ng/ml) revealed similar overall survival. Conclusively, preoperative concentration of serum VEGF may be the most valuable marker in the prediction of intratumoral angiogenesis and prognosis of patients with NSCLC. PMID- 15793666 TI - Pilot phase II study of gemcitabine and vinorelbine in patients with recurrent or refractory small cell lung cancer. AB - The aim of this phase II study was to evaluate toxicity, response, time to progression, and overall survival in patients with recurrent or progressive small cell lung cancer (SCLC) receiving the combination of gemcitabine and vinorelbine. This two stages Simon design trial was stopped after 17 patients were enrolled and response rate did not reach a level to justify continuation to the second stage. One patient had received three previous chemotherapy combinations, 5 had two prior chemotherapy regimens, and the remaining 11 had been treated with one prior line of therapy. Their median age was 62 years (35-72). Vinorelbine was administered at 25 mg/m(2) followed by gemcitabine 1000 mg/m(2), on days 1 and 8, every 3 weeks. Sixteen patients were evaluable for toxicity and 11 for response. Grade 3 neutropenia was seen in 5 patients (33%) and 3 patients had grade 4 neutropenia (20%). Three patients had grade 3 thrombocytopenia. No grade 4 non hematological toxicities were seen. A total of 85 infusions were given, with 1 patient (6%) obtaining a partial response (PR) for a duration of 148 days. Three additional patients had stable disease (19%), but only in 1 patient for longer than 24 weeks. The median time to progression was 47 days for all patients (range 25-196). After a median follow-up of 184 days, 13 patients died (76%). Median overall survival was 164 days. The combination of gemcitabine and vinorelbine has limited activity in relapsed SCLC. PMID- 15793667 TI - The influence of gastroesophageal reflux disease and its treatment on asthmatic cough. AB - Gastroesophageal reflux is known to cause chronic cough and is also implicated in worsening of asthma. We conducted a prospective study to examine the clinical significance of gastroesophageal reflux disease (GERD) in asthmatic patients with chronic cough to analyze the temporal relationship between reflux events and coughing and to assess the effect of esomeprazole treatment on respiratory symptoms and lung function in these patients. Asthmatic patients (126) with chronic dry cough were studied. Diagnosis of GERD was based on typical symptoms and the effectiveness of therapeutic test or on pH monitoring. Patients without GERD (negative pH results) consisted of the control group. The results of pH monitoring showed that 64% of cough episodes were related to acid reflux and in 91% of reflux events preceded coughing. Esomeprazole treatment (40 mg/day for 3 months) not only diminished GERD symptoms but also improved asthma outcome Baseline FEV(1) and PEF values increased significantly together with a decrease in symptom scores and the use of rescue medication. In most patients included in the extended part of the study for another 3 months, the dose of inhaled steroids could be reduced with sustained GERD therapy. Our data showing that reflux events preceded coughing in most cases and that treatment of GERD resulted in an improvement in different outcome measures of asthma suggest that GERD worsens asthma, and its treatment is of clinical importance to effectively manage these patients. PMID- 15793668 TI - Pharmacological study of oyster mushroom (Pleurotus ostreatus) extract on isolated guinea pig trachea smooth muscle. AB - Mushroom farm workers suffer from respiratory symptoms during the farming of mushrooms. The objective of this study was to analyze the effects of oyster mushroom (Pleurotus ostreatus) extract (OME) on isolated guinea pig tracheal smooth muscle in vitro. Isolated guinea pig tracheal tissue from 27 nonsensitized guinea pigs were studied. The OME was obtained from indoor mushroom growing fields and prepared as a 1:10 w/v aqueous solution. Dose-related contractions of nonsensitized guinea pig trachea were demonstrated using these extracts. The OME contained significant quantities of bacterial components (eg., endotoxin: 43,072.92 EU/mg). Parallel, pharmacological studies were performed by pre treating the tissues with mediator-modifying agents including atropine, indomethacin, pyrilamine, BPB, acivicin, NDGA, captopril, TMB8 and capsaicin. Atropine consistently and strikingly reduced the contractile effects of this extract. These observations suggest an interaction of the OME with parasympathetic nerves or more directly with muscarinic receptors. Pretreatment with TMB8 (inhibitor of intracellular calcium mobilization) also significantly blocked the constrictor effect of OME, indicating a role of calcium mobilization in the constricting effect of OME. Inhibition of contraction by blocking of other mediators was less effective and varied depending on the drug. We conclude that OME causes a dose-related airway smooth muscle constriction by nonimmunological mechanisms involving a variety of airway mediators and possibly cholinergic receptors. This effect is not dependent on pre-sensitization of the guinea pigs. PMID- 15793669 TI - Executive function improvement upon remission of recurrent unipolar depression. AB - The aim of the study was to investigate the improvement of executive function measures upon recovery from unipolar depression. Thirty patients who suffered from recurrent major unipolar depression were retested with regard to their executive function approximately two years after an initial baseline examination. At baseline, patients were depressed (average 17-item HAM-D score 21.8), at retesting they were partially or totally recovered (average HAM-D score 8.2). There was a significant positive association between improvement on the HAM-D and improvement of executive function. In those with complete recovery, overall executive function and most examined executive function measures were no longer different from the baseline performance of healthy controls (with the possible exception of semantic fluency and Stroop Colour-word). In conclusion, recovery from major unipolar depression was accompanied by a recovery of many aspects of executive function to a normal level. Our findings support previous studies that have shown that neuropsychological impairment associated with long-standing depressive symptomatology is reversible (i. e. state-related) in recurrent unipolar depression. PMID- 15793670 TI - Determinants of plasma adiponectin levels in patients with anorexia nervosa examined before and after weight gain. AB - OBJECTIVE: To examine the determinants of adiponectin levels (i) in 23 women with anorexia nervosa (mean BMI 15.0 +/- 1.2) and 43 healthy normal weight females (mean BMI 22.3 +/- 2.3; cross-sectional design) as well as (ii) after six and twelve weeks of weight gain in subgroups of 18 and 11 anorectic patients (mean weight gain 5.8 kg; longitudinal design). Plasma adiponectin and leptin concentrations were measured and their relationships to body composition (fat mass by bioelectrical impedance analysis and anthropometrics), different hormones and metabolic parameters (insulin, ACTH, cortisol, glucose, FFA, lipid profile) were investigated. RESULTS: In anorectic patients, adiponectin levels were higher (+29 %) and leptin levels were lower (-75 %) than in control subjects. There was a high variance in adiponectin levels in patients ranging from 2.6 to 18 nM. Combining patients and controls, an inverse linear correlation was observed between adiponectin levels and fat mass (r = -0.36, p < 0.05), while a positive exponential relation was found between leptin levels and fat mass (r = 0.82, p < 0.001). In anorectic patients, there were no significant correlations between adiponectin and hormonal or metabolic parameters. Weight gain resulted in increasing leptin (+0.17 +/- 0.12 nM; p < 0.001) and a nonsignificant decrease in adiponectin concentrations (-1.12 +/- 2.51 nM). Changes in leptin levels were mainly explained by a gain in fat mass (r = 0.85, p < 0.001). In contrast, changes in adiponectin levels were closely linked to initial adiponectin levels (r = -0.84, p < 0.001) but not to changes in fat mass or BMI. CONCLUSION: Cross sectionally serum adiponectin concentration followed a linear inverse function with fat mass when patients and controls were combined. Longitudinally gain in fat mass was not associated with changes in adiponectin levels suggesting other yet unidentified influences on adiponectin secretion in anorexia nervosa. PMID- 15793672 TI - [Position on choice of agents among TNF inhibitors]. PMID- 15793673 TI - [Recommendations of the German Society of Rheumatology on therapy of Raynaud syndrome and acral ulcerations]. PMID- 15793674 TI - [Recommendations of the German Society of Rheumatology on therapy of pulmonary hypertension in patients with autoimmune diseases]. PMID- 15793675 TI - [Treatment with leflunomide in rheumatoid arthritis]. PMID- 15793677 TI - [Lupus in Germany: analysis within the German lupus self-help organization (LULA)]. AB - During the last few decades, the prognosis for patients with systemic lupus erythematosus (LE) has changed from high early mortality to a more chronic longterm course. Although the prevalence of LE has been estimated at 20 50/100,000, data concerning the situation of LE patients in Germany are sparse. Since 2001, a documentation within the German Lupus Self-Help Organisation scheduled for a period of 10 years (LULA) has been recording at the patient level the actual status and the long-term course of a large group of LE patients. A questionnaire adapted from the German rheumatological database is updated once a year and sent to all members.In 2001, 1033 members participated in the documentation. Of these, 92.2% were women (mean age 45.8 years) with a mean disease duration of 9.9 years. 37.6% were employed, and 24.5% were on early retirement. 50.2% rated their overall health status as "not so good" or "poor". Most were receiving treatment with [hydroxy-]chloroquine (35.2%) or azathioprine (21.9%), while 67.9% were receiving corticosteroids. The most frequent comorbidities reported were hypertension (33%), scarring skin disease (24.4%), osteoarthritis (25.2%), osteoporosis (24%), psychiatric disorders/depression (22.9%) and chronic renal disease (22%). Thromboembolic events were reported in 18.5%, myocardial infarction in 2.3% and stroke in 4.8% of cases. Concerning their main contact person for health care, 63.6% specified the rheumatologist. In comparison with other cohort studies and in particular with the German rheumatological database, the data provided exclusively by patients are feasible. Concerning the severity of their disease, their treatment and their global assessment of health status, LULA participants are comparable with other LE patients and can be seen as representative of LE patients in Germany. Further assessment especially of long-term data are needed to obtain additional insights into the burden of the disease and the need for special medical care. PMID- 15793679 TI - [Is therapy with calcium and vitamin D adequate in glucocorticoid induced and inflammation related osteoporosis?]. PMID- 15793676 TI - [Intravenous immunoglobulins in chronic idiopathic myositis]. PMID- 15793678 TI - [Medicinal vasoactive therapy of microcirculation disorders in rheumatoid arthritis]. PMID- 15793680 TI - [Letter query to Zeitschrift fur Rheumatologie by Dr. med. Ilka Schwarze FA of Internal Medicine and Rheumatology Center of Internal Medicine--Leipzig University Clinic]. PMID- 15793684 TI - Behavioural characteristics and autistic features in individuals with Cohen Syndrome. AB - Diagnostic criteria for Cohen Syndrome are based largely on physical characteristics, and systematic information about behaviour and social functioning is limited. Typically, individuals with this condition are described as being very sociable and as showing low rates of pathology. However, recent studies have indicated that behavioural difficulties may occur more frequently than previously suggested and that autistic features may be relatively common. The present investigation of 45 individuals with Cohen Syndrome (age 4-48 years) found that, although 57% of the sample were reported as showing some behavioural disturbance, problems related mainly to anxiety and social interactions; marked anti-social behaviours were rare. Twenty-two individuals met criteria for autism on standardised diagnostic assessments, although the "autistic profile" was somewhat atypical. The implications of these findings for our understanding of Cohen Syndrome are discussed. PMID- 15793685 TI - What distinguishes autism spectrum disorders from other developmental disorders before the age of four years? AB - The increasing recognition of the benefits of early intervention for children with autism spectrum disorder (ASD) stresses the importance of early identification of children who might benefit from those programs. However, in the early years of life it may be difficult to distinguish children with ASD from children with other developmental disorders. The aim of the present study was to identify behavioural patterns that could facilitate this differentiation. Prior to diagnostic assessment, 2- and 3-year-old children (n=30), all referred to a clinic for "possible autism", were observed in a semi-structured play interaction, and their parents were interviewed about the children's early development from 0 to 24 months. Following diagnostic assessment, the 17 children fulfilling the ICD-10 criteria for ASD were compared to the 13 children diagnosed with other developmental disorders (outside the autism spectrum). On the basis of parent reports only a few distinguishing signs of ASD were found before 24 months of age. On the basis of professional observations in a semi-structured play interaction several distinguishing signs were found for the 2- and 3-year-olds; smiles in response, responds to name, follows pointing, looks to "read" faces, initiates requesting verbal and nonverbal behaviours, and functional play. PMID- 15793686 TI - The Strange Stories test--a replication study of children and adolescents with Asperger syndrome. AB - The aim of the present study was to assess the ability of 21 children and adolescents with Asperger syndrome (AS) of normal intelligence to infer mental states in a story context using Happe's Strange Stories test. The participants in the AS group were compared with an age-matched control group (N=20) of normally developing children and adolescents on a test of social understanding. The test material comprised social communication such as Pretence, Joke, Lie, White Lie, Figure of Speech, Misunderstanding, Persuasion, Irony, Double Bluff and Contrary Emotions, Appearance/Reality and Forgetting. As compared to the controls, the participants in the AS group performed less well on these tasks, and answered fewer correct mental state inferences, but performed well on a physical state control task. This study supports the main finding of earlier studies, showing that even individuals with AS of normal intelligence have problems in using mental state terms context-appropriately when tested on the Strange Stories test. PMID- 15793689 TI - A systematic review of age-related errors in children's memories for voiding cystourethrograms (VCUG). PMID- 15793687 TI - Stability and prediction of schizophrenia from adolescence to adulthood. AB - Stability of schizophrenia diagnosis from adolescence to adulthood, antecedents of schizophrenia, and differences in developmental and behavioural histories between subjects with early onset schizophrenia (EOS) and with adult onset schizophrenia (AOS) were investigated in 145 adult subjects diagnosed with mental disorders in adolescence and re-diagnosed on the basis of medical records according to DSM-IV.A very high diagnostic stability schizophrenia was demonstrated at the 28-year follow-up. Several factors, including neurological adversities, delayed language development, low IQ, and congenital functional disability, differentiated significantly between schizophrenic subjects and non schizophrenic subjects. Histories of concussion, physical abuse, parental divorce, and unstable familial context differentiated significantly between EOS and AOS subjects. Our findings support earlier evidence of schizophrenia being a chronic disorder with high diagnostic stability, and confirm the importance of neurological adversities, delayed language development, and low IQ as factors predictive of schizophrenia. Exploration of four case histories of AOS subjects delineates "pre-schizophrenic warning cluster" where combination of neurological adversities, temperamental problems, antisocial behaviour, preference for solitary play, and unstable family system constitute main factors. PMID- 15793688 TI - A risk and protective factors framework for understanding youth's externalizing problem behavior in two different cultural settings. AB - OBJECTIVE: The main goal of the study was to analyze youth's externalizing problem behaviors based on a risk and protective factors framework in two different cultural settings. METHOD: Data were collected among secondary school students in Szeged, Hungary (N=1240) and in Birmingham, AL, USA (N=1538). The self-administered questionnaires contained items on youth's externalizing problem behaviors as well as risk and protective factors. RESULTS: In both samples, first year students in secondary (high) schools and boys reported greater levels of problem behaviors. Multiple regression analyses revealed that substance use, gang membership and low academic achievement were consistent risk factors and associated with higher reporting levels of problem behaviors in both cultures. Parental monitoring served as an important protective factor in both samples, while school protective factors were only significant for American students. CONCLUSION: Findings draw attention to similar structures of certain risk and protective factors of youth's externalizing problem behaviors in different cultural settings. While there are universal risk factors (e. g., substance use, gang membership and low academic achievement), parental monitoring seems to be a universal protective factor against youth's externalizing problem behavior. An important difference is that the school domain seems to act as more important protection for American youth. PMID- 15793690 TI - Depression, anxiety, and obsessionality in long-term recovered patients with adolescent-onset anorexia nervosa. AB - Anorexia nervosa (AN) is frequently associated with symptoms of depression, anxiety, and obsessive-compulsive behavior which also develop secondary to semistarvation. It is less certain if these symptoms persist after recovery. A few studies have already reported on high prevalence rates of anxious, depressive, and obsessive features in long-term recovered patients with AN, but several of these so called "long-term" recovered patients had only maintained weight restoration for six to twelve months. The aim of this study was to determine whether depressive, anxious, and obsessive-compulsive symptoms persist in truly long-term recovered patients (BMI 20.3+/-2.5 kg/m(2)) who no longer had any eating disorder symptoms (including weight phobia) for at least 3 years. Seventeen subjects of an AN sample (n=39) previously described in a 10-year follow-up met our strict criteria of at least 3 years of complete recovery of AN. In comparison to 39 age-, sex-, and occupation-matched healthy subjects without a history of psychiatric or eating disorder, long-term recovered patients had higher levels of depressive (p=0.002), anxious (p=0.006), and obsessive compulsive (p=0.015) features but did not differ with regard to psychiatric morbidity and psychosocial adaptation. In conclusion, depressive, anxious, and obsessive-compulsive symptoms may be personality traits in subjects with former adolescent anorexia nervosa. PMID- 15793693 TI - [The diagnostic validity of screening tests and laboratory markers in alcohol use disorders]. AB - OBJECTIVE: To determine the validity of screening tests and markers suggested in the early detection of alcohol use disorders. METHOD: Male patients aged 20-65 were recruited from a psychiatry clinic. They had been consuming alcohol at least once a week. They were not drug users and had no other diseases that affected the markers mentioned in this study. According to the DSM-IV diagnostic criteria, the diagnostic validity of screening tests [Michigan Alcoholism Screening Test (MAST), CAGE (Cut down, Annoyed, Guilty, Eye-opener), Alcohol Use Disorders Identification Test (AUDIT)] and markers [Mean corpuscular volume (MCV), aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyltransferase (GGT), carbohydrate-deficient transferrin (CDT)] was investigated in alcohol dependent patients (n=29), alcohol abusers (n=28) and social drinkers (n=28). RESULTS: For the diagnosis of dependence, the sensitivity and specificity of the tests were respectively 69% and 100% for GGT, 41% and 93% for CDT, 100% and 57% for MAST, 97% and 86% for CAGE, and 100% and 75% for AUDIT. For the diagnosis of abuse, the sensitivity and specificity of the tests were respectively 32% and 100% for GGT, 32% and 93% for CDT, 86% and 57% for MAST, 75% and 85.7% for CAGE, and 89% and 75% for AUDIT. The widest area under the ROC curve belonged to GGT as a marker and AUDIT as a screening test. CONCLUSION: According to the results of this study, the marker that has the highest validity is GGT, while AUDIT and CAGE are the best screening tests. PMID- 15793694 TI - [Quality of life in patients with obsessive-compulsive disorder: relations with cognitive functions and clinical symptoms]. AB - OBJECTIVE: To compare quality of life in patients with obsessive-compulsive disorder with that in healthy subjects and to relate quality of life to cognitive functions and the severity of clinical symptoms. METHODS: Twenty-three patients who met DSM-IV criteria for obsessive compulsive disorder and 22 healthy subjects were included in the study. Quality of life (Turkish Quality of Life Scale-Brief Form) and cognitive functions were investigated in all subjects. In the patient group the relation of quality of life to the cognitive functions and to the severity of clinical symptoms and in the control group the relation of quality of life to the cognitive functions was investigated. RESULTS: The comparison of quality of life between the patient and control groups showed a significant difference (F= 2.60, p= 0.04). The significant differences between the two groups in psychological and social scores were responsible for the overall significant difference. The scores of quality of life were correlated with the scores of the cognitive tests (Trail Making Test, Auditory Consonant Trigram Test and Digit Span Test) and the severity of obsessive-compulsive symptoms. CONCLUSION: The present study revealed that quality of life is lower in patients with obsessive compulsive disorder than in healthy subjects and is related to cognitive functions and the severity of obsessive-compulsive symptoms. PMID- 15793695 TI - [Family functioning and psychosocial characteristics in children with attention deficit hyperactivity disorder with comorbid oppositional defiant disorder or conduct disorder]. AB - OBJECTIVE: To compare the parental sociodemographic characteristics, prenatal and postnatal developmental variables, IQ and behavioral disturbances as well as family functioning and current psychiatric disorders in the parents of children with attention deficit hyperactivity disorder (ADHD) and the parents of children with ADHD and comorbid oppositional defiant disorder (ODD) or conduct disorder (CD). METHOD: The sample consists of 92 children in the 6-11 age range, diagnosed with ADHD and ADHD with comorbid ODD/CD using DSM-IV diagnostic criteria. Parents completed the Child Behavior Checklist (CBCL) 4-18 and the Family Assessment Device (FAD) and were interviewed for current psychiatric treatment and alcohol consumption. RESULTS: 69.6% of the sample was diagnosed with ADHD and 30.4% with ADHD + ODD/CD. There were no differences between the two groups with respect to age, intelligence, characteristics of the neonatal period, age of walking and age of speech. Children with ADHD and comorbid ODD/CD had high CBCL subscale scores except for the social withdrawal and sexual problems subscales. Maternal depression and paternal drinking problems were high in the ADHD+ODD/CD group. The families of children with ADHD+ODD/CD also scored high at the level of 'unhealthy functioning' in the Roles and Behaviour Control subscales of the FAD. CONCLUSION: The treatment of children diagnosed with ADHD with comorbid ODD / CD should include parental treatment and intervention addressing parental skills. PMID- 15793696 TI - [Suicide probability: an assessment terms of reasons for living, hopelessness and loneliness]. AB - PURPOSE: The purpose of the current study is to specify the reasons that hold people clinging to life; to investigate their suicide probability, and to describe the relationship of these variables with other variables such as hopelessness and loneliness, taking age, education and other socioeconomic variables into consideration. METHOD: The subjects were 683, randomly chosen adolescents and adults between the ages of 15-65, residing in Ankara and Izmir. The assessment instruments were Reasons for Living Inventory, Suicide Probability Scale, UCLA Loneliness Scale, and Beck Hopelessness Scale. The data were analyzed using the SPSS for Windows 10.00. RESULTS: The analyses revealed that the group aged between 15-25 years reported fewer reasons for living, higher suicide probability, more hopelessness and loneliness, compared to older ages. Moreover, women reported more reasons for living, along with less loneliness and hopelessness. The regression analyses pointed out that age, education level, hopelessness, loneliness and reasons for living are predictive variables for suicide probability. CONCLUSION: As it was previously reported the current study also revealed that age is an important variable to be taken into consideration when suicide probability is being determined. In addition, was also found to be an important variable, at least for this Country. In parallel with the results of the studies in the relevant literature, reasons for living, hopelessness, and loneliness were found to be significant predictors of suicide probability. PMID- 15793697 TI - [A reliability study of the Turkish version of the mornings-evenings questionnaire]. AB - PURPOSE: To investigate the reliability of the Turkish version of the self assessment mornings-evenings questionnaire (MEQ). METHODS: A Turkish version of the MEQ, which was originally designed by Horne and Ostberg (1976), was given to 618 (291 women and 327 men) volunteers, aged 18 to 57 (23.0+/-5.4 years). Participants were recruited from the university student population and staff who were not night-shift workers. Exclusion criteria included sleep disorders and a history of medical and psychiatric diagnoses. The participants completed the questionnaire twice, 15-20 days apart. The reliability was assessed by Cronbach's and Pearson's r coefficient. RESULTS: MEQ scores were 51.9+/-9.0 (range 22-73) and 52.4+/-9.5 (range 21-72) for the 1st and 2nd applications, respectively, and did not differ significantly between the two applications. Of the 618 subjects interviewed, 149 were self-reported as morning types at the 1st application and 169 as morning types at the 2nd application; 79 as evening types at the 1st application and 87 as evening types at the 2nd application; and 390 as intermediate types at the 1st application and 362 as intermediate types at the 2nd application. Cronbach's a was 0.785 and 0.812 for the 1st and 2nd applications, respectively, and the test-retest reliability coefficient was 0.84 (p< 0.0001). DISCUSSION: The Turkish version of the MEQ shows high reliability. Nevertheless, further evaluation using different subject groups such as the elderly, athletes and shift workers is required to confirm the reliability and validity of the Turkish version. PMID- 15793698 TI - [Impulsivity and mental disorders]. AB - OBJECTIVE: Although there are several studies revealing the role of impulsivity in mental disorders, debates in the literature about how to define and determine impulsivity continue. Being a measurable feature of behaviour, impulsivity is defined as the failure to resist a drive or stimulus or as a personality dimension as the inability to resist the desire to harm one's self or others. Besides this negative definition, however, impulsivity is also defined as independent dysfunctional and functional concepts having both positive and negative consequences. It can be a psychopathological structural part of many mental disorders, but it can be a characteristic of normal behaviour as well. Although not defined in detail in DSM-IV, it is mentioned as a diagnostic criterion in several mental disorders like impulse control disorders (pathological gambling, intermittent explosive disorder, pyromania, kleptomania and trichotillomania), impulsive aggressive disorders of personality (borderline, antisocial, histrionic and narcissistic), manic episodes of bipolar disorder, attention deficit hyperactivity disorder (ADHD), neurological disorders with behavioural disinhibition and substance abuse. Studies have revealed impulsivity to be more common in subjects with conduct disorders, attention deficit hyperactivity disorder, disorders of personality, substance and alcohol abuse, psychotic disorders, bipolar disorders, eating disorders and dementia compared to healthy subjects in control groups. The aim of the present paper is to define impulsivity as a concept and symptom, to discuss its relation to mental disorders, to show the biological basis of this relationship and to review the literature relating to this issue. PMID- 15793699 TI - [Creutzfeldt-Jakob disease: a case that initiated with psychiatric symptoms]. AB - Creutzfeldt-Jakob disease (CJD) is a rapidly progressive prion disease that causes deficits involving movement, cognition, and mental status. The clinical heterogeneity of the disease can make diagnosis difficult. Thorough neurologic, cognitive, and psychiatric examinations are necessary for observing its clinical features. In this case report we describe a 62-year-old male patient who was initially followed with a diagnosis of depression and later was diagnosed with CJD. The patient had a one-year history of anhedonia, loss of interest, social withdrawal, anxiety and decrease in speech and was given paroxetin 20 mg/day with a diagnosis of depression. During follow up, neurological symptoms including ataxia and rigidity became evident and dementia and akinetic mutism developed in a rapidly progressive course. Although electroencephalography (EEG) and magnetic resonance imaging (MRI) revealed nonspecific findings initially, typical findings for CJD were seen during the follow up. The positive 14-3-3 protein in CSF supported the diagnosis. The aim of this report is to emphasize the fact that CJD may present with different psychiatric symptoms and can be initially misdiagnosed. CJD should be considered in the differential diagnosis of patients who have focal neurological signs in addition to psychiatric symptoms. Repeated neurological examinations, EEG and cranial MRI may help in the diagnosis of these patients. PMID- 15793700 TI - [Charles Bonnet syndrome: a case presentation]. AB - Charles Bonnet syndrome comprises the triad of visual hallucinations, visual sensory deprivation, and preserved cognitive status. This paper discusses a case diagnosed as Charles Bonnet syndrome, involving visual hallucinations secondary to bilateral primary optic atrophy. An 80-year-old female with normal cognitive functions in the presence of primary optic atrophy and visual hallucinations was diagnosed with Charles Bonnet syndrome. The patient, having had poor vision since childhood, had lost it totally in the last year. Her vision had not improved following cataract operations in both eyes 6 months previously. Her vision was at the level of hand movements. In biomicroscopic examination, bilateral pseudoaphakia was found. Since fundus examination showed bilateral primary optic atrophy in the presence of visual hallucinations, a psychiatric consultation was requested. In her psychiatric examination, she had had hallucinations for the last two years, first elementary and then complex in character. Her cognitive functions were normal with no pathology in her neurologic examination. Routine investigations and neuroradiologic examinations were normal. She had no past history of any personal or familial psychiatric or systemic physical disorder. She was given olanzapine 5 mg daily and was followed up. This syndrome, defined as visual hallucinations in the presence of preserved cognitive functions and deprived vision, requires further research. PMID- 15793701 TI - The Epsin 4 gene on chromosome 5q, which encodes the clathrin-associated protein enthoprotin, is involved in the genetic susceptibility to schizophrenia. AB - Chromosome 5q33 is a region that has previously shown good evidence of linkage to schizophrenia, with four LOD scores >3.00 in independent linkage studies. We studied 450 unrelated white English, Irish, Welsh, and Scottish research subjects with schizophrenia and 450 ancestrally matched supernormal controls. Four adjacent markers at the 5' end of the Epsin 4 gene showed significant evidence of linkage disequilibrium with schizophrenia. These included two microsatellite markers, D5S1403 (P=.01) and AAAT11 (P=.009), and two single-nucleotide polymorphism markers within the Epsin 4 gene, rs10046055 (P=.007) and rs254664 (P=.01). A series of different two- and three-marker haplotypes were also significantly associated with schizophrenia, as confirmed with a permutation test (HapA, P=.004; HapB, P=.0005; HapC, P=.007; and HapD, P=.01). The Epsin 4 gene encodes the clathrin-associated protein enthoprotin, which has a role in transport and stability of neurotransmitter vesicles at the synapses and within neurons. A genetically determined abnormality in the structure, function, or expression of enthoprotin is likely to be responsible for genetic susceptibility to a subtype of schizophrenia on chromosome 5q33.3. PMID- 15793703 TI - The dual origin of the Malagasy in Island Southeast Asia and East Africa: evidence from maternal and paternal lineages. AB - Linguistic and archaeological evidence about the origins of the Malagasy, the indigenous peoples of Madagascar, points to mixed African and Indonesian ancestry. By contrast, genetic evidence about the origins of the Malagasy has hitherto remained partial and imprecise. We defined 26 Y-chromosomal lineages by typing 44 Y-chromosomal polymorphisms in 362 males from four different ethnic groups from Madagascar and 10 potential ancestral populations in Island Southeast Asia and the Pacific. We also compared mitochondrial sequence diversity in the Malagasy with a manually curated database of 19,371 hypervariable segment I sequences, incorporating both published and unpublished data. We could attribute every maternal and paternal lineage found in the Malagasy to a likely geographic origin. Here, we demonstrate approximately equal African and Indonesian contributions to both paternal and maternal Malagasy lineages. The most likely origin of the Asia-derived paternal lineages found in the Malagasy is Borneo. This agrees strikingly with the linguistic evidence that the languages spoken around the Barito River in southern Borneo are the closest extant relatives of Malagasy languages. As a result of their equally balanced admixed ancestry, the Malagasy may represent an ideal population in which to identify loci underlying complex traits of both anthropological and medical interest. PMID- 15793704 TI - Isolated imperforate anus in monozygotic twins: case report and implications. AB - The authors report a case of isolated imperforate anus with perineal fistula in monozygotic twins. Only 4 other well-documented cases of monozygotic twins concordant for isolated anorectal malformation have been reported in the literature. In all these cases the defect occurred below the levator ani muscle. This case confirms previous reports on the incidence of low anorectal anomalies in monozygotic twins where the defect was isolated. A review of the literature relating to this condition in twins and siblings points toward low anomalies and more severe malformations having different genetic backgrounds. PMID- 15793705 TI - Esophageal stenosis in infants: unusual complication after Pseudomonas aeruginosa sepsis. AB - The authors report 2 cases of 4- and 5-month-old male infants who had esophageal stenosis after Pseudomonas aeruginosa sepsis. These 2 babies were immunocompetent. A focal narrowing of esophagus at the level of C7-T1 was found on esophagogram. PMID- 15793706 TI - Surgical management for posterior laryngeal cleft developing subglottic airway obstruction. AB - A 3-year-old boy associated with Opitz syndrome including hypospadias and anorectal anomaly was transferred to Osaka University Hospital with orotracheal intubation because of respiratory distress after respiratory infection. He had laryngeal cleft diagnosed in the neonatal period because of symptoms including stridor and aspiration. After 2 attempts at extubation, tracheostomy was performed. Fiberscopic examination demonstrated obstruction of the airway from the vocal cord to subglottic space by inspired esophageal redundant mucosa. There was no improvement observed for 3 months. He underwent anterior repair of cleft, anterior laryngotracheoplasty with costal cartilage graft, closure of tracheostomy, and endoscopic Nissen fundoplication. The postoperative course during the subsequent year was uneventful without feeding or speaking disorders. Subglottic airway obstruction caused by inspired redundant mucosa is a rare complication in laryngeal cleft. Anterior laryngotracheoplasty and Nissen fundoplication should be considered for this complication, in addition to the usual procedure for cleft repair. PMID- 15793702 TI - Diversity and function of mutations in p450 oxidoreductase in patients with Antley-Bixler syndrome and disordered steroidogenesis. AB - P450 oxidoreductase (POR) is the obligatory flavoprotein intermediate that transfers electrons from reduced nicotinamide adenine dinucleotide phosphate (NADPH) to all microsomal cytochrome P450 enzymes. Although mouse Por gene ablation causes embryonic lethality, POR missense mutations cause disordered steroidogenesis, ambiguous genitalia, and Antley-Bixler syndrome (ABS), which has also been attributed to fibroblast growth factor receptor 2 (FGFR2) mutations. We sequenced the POR gene and FGFR2 exons 8 and 10 in 32 individuals with ABS and/or hormonal findings that suggested POR deficiency. POR and FGFR2 mutations segregated completely. Fifteen patients carried POR mutations on both alleles, 4 carried mutations on only one allele, 10 carried FGFR2 or FGFR3 mutations, and 3 patients carried no mutations. The 34 affected POR alleles included 10 with A287P (all from whites) and 7 with R457H (four Japanese, one African, two whites); 17 of the 34 alleles carried 16 "private" mutations, including 9 missense and 7 frameshift mutations. These 11 missense mutations, plus 10 others found in databases or reported elsewhere, were recreated by site-directed mutagenesis and were assessed by four assays: reduction of cytochrome c, oxidation of NADPH, support of 17alpha-hydroxylase activity, and support of 17,20 lyase using human P450c17. Assays that were based on cytochrome c, which is not a physiologic substrate for POR, correlated poorly with clinical phenotype, but assays that were based on POR's support of catalysis by P450c17--the enzyme most closely associated with the hormonal phenotype--provided an excellent genotype/phenotype correlation. Our large survey of patients with ABS shows that individuals with an ABS-like phenotype and normal steroidogenesis have FGFR mutations, whereas those with ambiguous genitalia and disordered steroidogenesis should be recognized as having a distinct new disease: POR deficiency. PMID- 15793707 TI - Assessment of obstructive apnea by using polysomnography and surgical treatment in patients with Beckwith-Wiedemann syndrome. AB - PURPOSE: Obstructive apnea is sometimes seen in patients with Beckwith-Wiedemann syndrome. The cause of apnea is not limited to macroglossia, and the surgical indication for obstructive apnea has not yet been established. The authors performed polysomnography for the assessment of apnea. METHOD: Overnight polysomnograms were obtained in 2 patients who developed obstructive apnea after 1-stage repair for omphalocele. CASE 1: Apnea index (AI), defined as apneic events per hour, indicated 17.3, and SpO2 below 95% occupied 80% of the total sleep time. Computed tomography and magnetic resonance imaging indicated obstruction of the airway between macroglossia and the hypopharynx. Central tongue resection and the division of the frenulum linguae for associated ankyloglossia were performed 97 days after birth. One month after surgery, apneic events disappeared and SpO2 below 95% occupied only 1% of the total sleep time. CASE 2: Obstructive AI indicated 28.1. Division of the frenulum linguae and anterior glossopexy were performed 55 days after birth. Postoperative polysomnogram indicated a marked reduction of AI. CONCLUSIONS: These results indicated that polysomnography was useful for evaluating obstructive apnea and that advancement of the tongue by division of the frenulum linguae may be recommended for the treatment of obstructive apnea in patients with Beckwith Wiedemann syndrome. PMID- 15793708 TI - Focal nodular hyperplasia of the liver: usefulness of superparamagnetic iron oxide-enhanced magnetic resonance imaging. AB - This report describes the use of superparamagnetic iron oxide-enhanced magnetic resonance imaging to diagnose fibronodular hyperplasia of the liver in a 12-year old girl. PMID- 15793709 TI - Obstruction of the proximal jejunum by an anomalous congenital band--a case report. AB - A case of proximal jejunal obstruction by a congenital band is reported. The band ran from the antimesenteric wall of the proximal jejunum just distal of the Treitz's ligament to the root of the mesentery. Lysis of the band and enterotomy were performed to make sure no intrinsic obstruction cured the patient. No recurrence was found after 3 months of follow-up. To the best of the authors' knowledge, this is the second case with an anomalous congenital band causing proximal jejunal obstruction reported in the English-language literature. PMID- 15793710 TI - Serial transverse enteroplasty as primary therapy for neonates with proximal jejunal atresia. AB - Small bowel atresia is associated with a large size discrepancy between the proximal and distal segments of bowel that has traditionally been managed by resection of the dilated segment, tapering enteroplasty, or plication. Longitudinal intestinal lengthening is rarely performed at the time of the initial operation. Many patients with small bowel atresia also have a short length of residual small intestine secondary to in utero resorption. The authors present the clinical application of the novel intestinal lengthening procedure, the serial transverse enteroplasty, in a neonate with proximal jejunal atresia and suggest that it become part of the armamentarium for surgeons treating patients with this anomaly. PMID- 15793711 TI - Living-donor liver transplantation for situs inversus: 2 case reports. AB - Two cases of living-donor liver transplantation performed in patients with situs inversus are reported. The authors discuss the operative management for a situs inversus recipient to undergo liver transplantation. PMID- 15793712 TI - Unusual case of epigastric heteropagus twinning. AB - Asymmetrical conjoined twins or heteropagus twins are extremely rare. They are characterized by an incomplete component (parasite) that is normally smaller and dependent on the host (autosite). In cases of an epigastric heteropagus twin, the insertion occurs in the epigastrium. There are few reports of epigastric heteropagus twinning in the English-language literature. The authors report an extremely rare case of epigastric heteropagus twinning in which the parasite presented with head, thorax, and a rudimentary heart. PMID- 15793713 TI - Application of memory metallic stents to urinary tract disorders in pediatric patients. AB - The use of memory metallic stents for the urinary tract in pediatric patients has not been reported. The authors report on 2 patients with urinary tract disorders who were successfully treated with a memory metallic stent. A thermoexpandable, nickel-titanium alloy stent was placed at the urethroureteral junction of a 4 year-old boy with ureteral stenosis associated with cloacal exstrophy for 18 months and at the urethra of a 2-year-old girl with ischuria after a repair of cloacal anomaly for 6 months. Temporary insertion of a memory metallic stent is a safe and effective alternative for organic stricture or functional obstruction of the urinary tract in pediatric patients. PMID- 15793714 TI - Remission induced by interferon alfa in a patient with massive osteolysis and extension of lymph-hemangiomatosis: a severe case of Gorham-Stout syndrome. AB - The treatment of massive osteolysis with lymphangioma and/or hemangioma (Gorham Stout syndrome) has been controversial. The authors report on a patient with multiple massive osteolyses and extensive lymph-hemangiomatosis whose lesions were reduced by interferon alfa therapy. A 2-year-old girl had complained of left chylothorax. Thoracoscopy showed an increase in small lymphatic vessels in the chest wall. The chylothorax was improved by coagulation of the lymphatic vessels. Later, multiple massive osteolyses appeared in the left 11th and 12th ribs, the TH10-L3 vertebrae, and the right femur. There were also hemangiomas in the liver and spleen, a tumor lesion in the left lower chest wall, and hemangiomatous change on the skin surface of the left back. The left lung had only a minimal air content. After OK-432 was injected into the femur and chest wall lesions, the femur lesion disappeared. Then, as right chylothorax appeared, OK-432 was injected into the right pulmonary cavity. The chylothorax disappeared, but pericardial effusion appeared. After steroid pulse therapy, pericardial effusion disappeared. During these treatments, the 7th to 10th ribs disappeared from the x ray and scoliosis developed. One month later, a cloudy fluid collection in the right lung was found on computed tomography. Interferon alfa and steroid pulse therapy were started. Interferon alfa (1,500,000 units) was subcutaneously administered daily for 2 months and was gradually reduced and maintained at 1,500,000 unit/wk. Steroids were also reduced and maintained at 5 mg/d of predonine. Later, the progress of osteolysis and the extension of lymph hemangiomatosis stopped. Ten months later, hemangioma in the back disappeared, and the 7th to 10th ribs, which had disappeared, reappeared. The interferon alfa therapy was stopped 14 months after it was administered. The patient's condition has been stable for 10 months since then. At this time, computed tomography shows regression of the hemangiomatous lesion in the back. The authors clinically diagnosed the patient as having Gorham-Stout syndrome with extension of lymph hemangiomatosis. Interferon alfa with or without steroid therapy should be a choice for patients with extension lesions. PMID- 15793715 TI - An exceptional combined malformation: duplication of the urinary and intestinal tracts and the vulva (04-80CR). AB - The authors reported the surgical treatment of a 2-year-old girl with complete duplication of the kidney, ureter, bladder, urethra, and the vulva, associated with intestinal duplication and complete duplication of the appendix and colon. Congenital intestinal malrotation also concurred. After a series of preoperative examinations, exploratory operation including reduction of intestinal malrotation, fusion of bladder and colon, obliteration of the duplicated urethra, resection of the intestinal and appendiceal duplications, and cosmetic repair of the vulva was performed. Incontinence of urine and stool disappeared, and she had been followed up for 10 months until this report. Repeat voiding cystourethrography performed recently demonstrated a fused bladder and the disappearance of the duplicated urethra. PMID- 15793716 TI - Usefulness of selective contrast echocardiography and selective scintigraphy for the evaluation of pulmonary arteriovenous fistula in a patient with systemic arterial supply to a normal lung. AB - To determine a surgical procedure for the correction of a systemic arterial supply to a normal lung, it is important to evaluate whether the patient has a pulmonary arteriovenous fistula. In this article, the authors report for the first time a patient in which selective contrast echocardiography and selective perfusion scintigraphy from an abnormal artery clearly demonstrated and quantified the pulmonary arteriovenous fistula. The authors believe that these methods are useful in determining the appropriate operative procedure for this condition. PMID- 15793717 TI - Torsion of an ectopic liver in a young child. AB - A large ectopic liver nodule connected to the greater omentum had undergone infarction and separation because of torsion of its connecting stalk and was the cause of acute abdominal pain and elevated liver enzymes in a young female child. At laparotomy for a suspected perforated appendicitis, a hemorrhagic peritoneal fluid was found, together with an intraperitoneal solid mass lying freely in the right iliac fossa. The definitive diagnosis was obtained by histopathology. This is the first known such reported case. PMID- 15793718 TI - Insulinlike growth factor I gene expression is increased in the fetal lung after tracheal ligation. AB - BACKGROUND/PURPOSE: The mortality and morbidity in congenital diaphragmatic hernia are mainly caused by pulmonary hypoplasia. To improve clinical results, further methods inducing lung growth may have to be used. The aim of this report was to evaluate the expression of insulinlike growth factor I (IGF-I), estrogen receptor alpha, estrogen receptor beta, growth hormone receptor, and thioredoxin in a rat model of hypoplastic, hyperplastic, and normal fetal lungs to improve understanding of lung growth. METHODS: Hypoplastic diaphragmatic hernia lungs were created by giving nitrofen by gavage to pregnant rats on day 9.5. Hyperplastic lungs were achieved by intrauterine tracheal ligation of rat fetuses on day 19. All lungs were harvested on gestational day 21. Total nucleic acids were extracted by proteinase K digestion and extraction in phenol/chloroform. The total nucleic acids mixture was hybridized with radioactively labeled RNA probes, and the radioactivity of the hybrids was compared with the respective standard curve of known amounts of in vitro synthesized mRNA. Immunohistochemistry staining was performed for IGF-I. RESULTS: The IGF-I mRNA was significantly (P < .01) higher in hyperplastic lungs compared with control and hypoplastic lungs. The latter 2 did not differ. No difference was found between the other mRNA levels in the study groups. CONCLUSIONS: IGF-I is involved in the accelerated lung growth seen after intrauterine tracheal ligation. PMID- 15793719 TI - Altered chemokine response in an animal model of multiple organ dysfunction syndrome induced by zymosan. AB - PURPOSE: The aim of this study was to determine the monocyte chemoattractant protein-1 (MCP-1) response over time in an animal model of multiple organ dysfunction syndrome (MODS). METHODS: On day 0, rats were randomized to receive an intraperitoneal injection of zymosan at a dose of 1 mg/g of body weight (n = 36) or vehicle (n = 9). Serum, peritoneal lavage (PL) fluid, and bronchoalveolar lavage (BAL) fluid were collected from 3 rats in the control group and 6 to 7 rats in the zymosan group at days 1, 5, and 12. Monocyte chemoattractant protein 1 concentrations were determined by enzyme-linked immunosorbent assay. RESULTS: The authors observed a 47% mortality in the zymosan-treated rats. Monocyte chemoattractant protein-1 levels were unchanged in the serum, PL, and BAL of control animals. Both serum and PL MCP-1 were significantly higher in zymosan treated rats on days 1 (P < .01) and 5 (P < .05) when compared with controls. By day 12, no difference between the 2 groups was observed. No significant difference was noted in BAL MCP-1. CONCLUSIONS: Chemokines are increased systemically and locally during MODS. The fact that MCP-1 is significantly higher early in the course of MODS may suggest that this chemokine is important in the early inflammatory changes that lead to MODS later in the course of this illness. PMID- 15793720 TI - Ischemia/reperfusion: a clinically relevant model of intestinal injury yielding systemic inflammation. AB - BACKGROUND/PURPOSE: Multisystem organ failure (MSOF) is a major cause of morbidity and mortality in the critically ill patient. Animal models of endotoxin induced sepsis were used to develop therapeutic regimens, which thus far have failed in clinical trials. Because multiple etiologies of MSOF affect the intestine, the authors hypothesized that during sepsis the gut may act as a possible trigger of the inflammatory cascade. As ischemia and reperfusion of the small intestine disrupts gut barrier function, thereby activating systemic inflammatory responses, the authors evaluated a murine model of ischemia/reperfusion to investigate these systemic responses to local mucosal and epithelial injury. METHODS: C57BL/10 and Balb/c mice underwent variable amounts of gut ischemia by superior mesenteric artery occlusion. Animals were evaluated for survival as well as gross and microscopic intestinal damage. RESULTS: Maximal ischemic damage occurred in the distal jejunum and proximal ileum. More severe epithelial damage and transmural inflammation were observed in C57BL/10 mice, which correlated with a higher mortality. CONCLUSIONS: This model mimics what is observed clinically with intestinal injury resulting from a progressive ischemic insult with eventual systemic manifestations. This reproducible model of systemic inflammation elicits variable responses from genetically different animals, the results of which may lead to a better understanding of MSOF. PMID- 15793721 TI - Radionuclear transit to assess sites of delay in large bowel transit in children with chronic idiopathic constipation. AB - PURPOSE: Chronic idiopathic constipation (CIC) with soiling in children may result from slow colonic transit (SCT) or anorectal dysfunction and/or psychological problems known as functional fecal retention (FFR). Evidence is accumulating that SCT and FFR need different treatments, but they are poorly distinguished by solid marker studies. The authors used radionuclear transit scintigraphy to categorize children with CIC as having either FFR or SCT. METHODS: Children (N = 101) with CIC (and soiling) who were referred for further investigation after failure of standard treatments (diet, laxatives) received radiolabeled colloid orally, and scintillation images were collected at 0 to 2, 6, 24, 30 and 48 hours (total radiation dosage = 2 standard x-rays). Radioactivity in 6 regions (precolonic, ascending, transverse, descending, rectosigmoid, and evacuated feces) was measured, and the median position (geometric center) of radioactivity at each time was determined. RESULTS: In children, meals normally reach the cecum at 6 hours and are evacuated in 30 to 58 hours. Fifty patients had retention of radioactivity in the proximal colon at 48 hours, indicating SCT. Analysis of the images and the geometric center showed that passage through the ascending colon and transverse colon was delayed in SCT. In 24 patients, radioactivity was passed by 30 hours, indicating normal transit or possible FFR. Twenty-two patients had retention in the rectum, indicating definite FFR. Five studies were borderline. CONCLUSIONS: Radionuclear transit scintigraphy is useful for categorizing patients with CIC as having either FFR or SCT, allowing for different treatments. Radionuclear transit scintigraphy provides more detail and greater sensitivity than solid marker studies in diagnosing CIC. Radionuclear transit scintigraphy showed that half of our patients had SCT. PMID- 15793722 TI - Does maternal nicotine exposure during gestation increase the injury severity of small intestine in the newborn rats subjected to experimental necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: The aim of this study was to evaluate the effects of maternal nicotine exposure during gestation on injury severity of small intestine in the newborn rats subjected to hypoxia-reoxygenation and cold stress. METHODS: A total of 21 Sprague-Dawley pregnant rats were divided into 3 equal groups. The groups were labeled as group 1, control group; group 2, hypoxia-reoxygenation group; and group 3, nicotine-hypoxia-reoxygenation group. The rats of group 3 were exposed to nicotine via subcuticular injection for the last week of gestation (2 mg/kg/d). Newborn rats were collected immediately after birth to prevent suckling of maternal milk (40 rat pups in group 1, 43 rat pups in group 2, and 41 rat pups in group 3). Litters in groups 2 and 3 were stressed twice daily with asphyxia followed by cold (4 degrees C for 10 minutes) stress to induce hypoxic intestinal injury which is relevant to human necrotizing enterocolitis. Breathing 100% CO2 for 10 minutes in a chamber followed by 10-minute 100% O2 breathing was the asphyxia model repeated twice daily. After hypoxia-reoxygenation and cold stress, newborn rats were returned to their mother's cages. This protocol was repeated for the following 2 days, and the rat pups were decapitated on the third day. Using this protocol of asphyxia and cold stress, all of neonatal rats developed clinical and pathological signs of hypoxia-induced intestinal injury. The entire gastrointestinal tract was removed and examined macroscopically. A 2-cm section of distal ileum from each animal was taken for histopathological and biochemical examinations. Histological changes in ileal architecture were scored and graded from 1 to 5. The remaining intestinal tissues of the animals were used for lipid peroxidation analysis. RESULTS: Typical signs of hypoxia-induced intestinal injury were observed in the 2 experimental groups (groups 2 and 3) macroscopically. There were more grades 3 and 4 injuries in group 3 (P < .05). The malondialdehyde levels were elevated in groups 2 and 3 (P < .001). The malondialdehyde levels of the group 3 were also significantly higher than group 2 (P < .01). CONCLUSIONS: Maternal nicotine exposure during gestation results in higher grade histological injury in newborn rats subjected to hypoxia reoxygenation and cold stress. PMID- 15793723 TI - Effect of sex and sex hormones on structural intestinal adaptation after massive small bowel resection in rats. AB - PURPOSE: The gonadal steroids play a major role in the regulation of many functions. The purpose of the current study was to evaluate the effect of sex and sex hormones on intestinal adaptation in a rat model of short bowel syndrome (SBS). METHODS: In the first experiment, male and female Sprague-Dawley rats underwent bowel transection and re-anastomosis (sham group) or 75% small bowel resection and anastomosis (SBS group). Relative changes in parameters of intestinal adaptation (overall bowel and mucosal weight, mucosal DNA and protein, villus height, and crypt depth) were measured on day 15 and were compared with respect to sex. In the second experiment, male rats were divided into 4 experimental groups: SBS rats, SBS castrated rats, SBS castrated rats treated with testosterone, and SBS castrated rats treated with estradiol. Parameters of intestinal adaptation were compared with respect to hormonal treatment. Statistical significance was determined by Student's t test and analysis of variance with P < .05 considered significant. RESULTS: Sex had minimal effects on intestinal adaptation. Both male and female rats showed a comparable increase in all parameters of intestinal adaptation. In the second experiment, castration led to significant decrease in bowel and mucosal weight, mucosal DNA and protein in both jejunum and ileum compared with SBS animals. Castrated rats also had lower jejunal villus height and crypt depth compared with SBS animals. Testosterone attenuated this negative effect of castration on bowel regrowth. Rats treated with testosterone showed a significant increase in bowel and mucosal weight, mucosal protein in both jejunum and ileum, mucosal DNA, villus height, and crypt depth in jejunum compared with castrated nontreated animals. Treatment with estradiol after resection and castration had minimal effect on bowel regrowth. CONCLUSIONS: Bowel regrowth after massive small bowel resection is not sex related. Depletion of androgens by castration inhibited intestinal adaptation. Testosterone has shown a strong stimulating effect on bowel regrowth. PMID- 15793724 TI - The vacuum chest wall lifter: an innovative, nonsurgical addition to the management of pectus excavatum. AB - PURPOSE: This report describes the authors' experience using a vacuum to pull the abnormal chest wall outward in patients with pectus excavatum. METHODS: A suction cup was used to create a vacuum at the chest wall. A patient-activated hand pump was used to reduce pressure up to 15% below atmospheric pressure. The device was used by 60 patients (56 males, 4 females), aged 6.1 to 34.9 years (median, 14.8 years), for a minimum of 30 minutes, twice a day, up to 5 hours per day (median, 90 minutes). Patient progress was documented using photography, radiography, and plaster casts of the defect. In 14 children this method was used during the Nuss procedure to enlarge the retrosternal space for safer passage of the introducer. RESULTS: Follow-up occurred between 2 and 18 months (median, 10 months). Computed tomographic scans showed that the device lifted the sternum and ribs within 1 to 2 minutes; this was confirmed thoracoscopically during the Nuss procedure. The suction cup enlarged the retrosternal space for safer passage of the introducer. Initially, the sternum sank back after few minutes. After 1 month, an elevation of 1 cm was noted in 85% of the patients. After 5 months, the sternum was lifted to a normal level in 12 patients (20%) when evaluated immediately after using the suction cup. All patients exhibited moderate subcutaneous hematoma, although the skin was not injured. One patient suffered from transient paresthesis in the right arm and leg. Two patients experienced orthostatic disturbances during the first application of the suction cup. There were no other complications. DISCUSSION: In patients with pectus excavatum, application of a vacuum effectively pulled the depressed anterior chest wall forward. The initial results proved dramatic, although it is not yet known how much time is required for long term correction. CONCLUSIONS: This vacuum method holds promise as a valuable adjunct treatment in both surgical and nonsurgical correction of pectus excavatum. PMID- 15793726 TI - Trimetazidine reduces the degree of fibrosis in alkali burns of the esophagus. AB - PURPOSE: The aim of this study was to investigate the efficacy of trimetazidine (TMZ), an antioxidant agent, on the prevention of stricture development after esophageal caustic injuries in rat. METHODS: Thirty rats were divided into 3 equal groups. A standard esophageal caustic burn was produced by application of 37.5% NaOH for a period of 90 seconds followed by water rinse. Group A (sham) animals were uninjured. Group B rats were injured but untreated. Group C rats were injured and received TMZ (5 mg/kg/d) via intraperitoneal route. Efficacy of the treatment was assessed in 28 days by measuring stenosis index and histopathologic damage score and by determining tissue hydroxyproline content. RESULTS: The stenosis index in the TMZ-treated group was significantly lower than the untreated group, similarly in the sham laparotomy group (stenosis index: 0.34 +/- 0.10, 0.94 +/- 0.21, 0.38 +/- 0.05, respectively; P < .05). The hydroxyproline level (microgram per milligram of wet tissue) was significantly lower in the TMZ-treated group compared with untreated group, similarly in the sham laparotomy group (1.06 +/- 0.14, 1.33 +/- 0.08, 0.68 +/- 0.15 microg/mg wet tissue, respectively; P < .05). In the untreated group, histopathologic damage score was significantly higher than TMZ-treated group (P < .05). CONCLUSIONS: Trimetazidine reduces the degree of fibrosis and ameliorates histopathologic damage in experimental model of corrosive esophagitis in rats. PMID- 15793725 TI - The use of a retrievable self-expanding stent in treating childhood benign esophageal strictures. AB - BACKGROUND/PURPOSE: Esophageal stenting is a popular form of treatment of esophageal strictures in adults but is not widely used in children. The aim of the current study was to investigate whether esophageal stents could be used safely and effectively in the treatment of esophageal stenosis in children. METHODS: Covered retrievable expandable nitinol stents were placed in 8 children with corrosive esophageal stenosis. The stents were removed 1 to 4 weeks after insertion. RESULTS: The stents were placed in all patients without complications and were later removed successfully. After stent placement, all patients could take solid food without dysphagia. Stent migration occurred in one patient and so the insertion procedure was repeated to reposition the stent. During the 3-month follow-up period after stent removal, all children could eat satisfactorily. After 6 months, 2 children required balloon dilation (3 times in one and 5 times in the other). The dysphagia score improved in all patients. CONCLUSIONS: The use of the covered retrievable expandable stent is an effective and safe method in treating childhood corrosive esophageal stenosis. PMID- 15793727 TI - Does antegrade cerebral perfusion protect the brain during deep hypothermic circulatory arrest? AB - PURPOSE: This study compares cerebral protection using no cerebroplegia and using antegrade cerebroplegia with variable flow rates during deep hypothermic circulatory arrest (DHCA). METHODS: Twenty healthy neonatal piglets (2.5-3.8 kg) underwent 60 minutes of DHCA. No cerebroplegia was used in group 1 (n = 5). Cold (16 degrees C) antegrade cerebral perfusate was administered through the innominate artery at 10 mL/kg per minute in group 2 (n = 5), at 25 mL/kg per minute in group 3 (n = 5), and at 50 mL/kg per minute in group 4 (n = 5). Venous samples for lactate, pyruvate, S-100B protein, and creatine kinase BB (CKBB) were drawn from the jugular vein before and after discontinuation of cardiopulmonary bypass--lactate at 5 minutes postbypass, pyruvate at 5 minutes postbypass, S-100B protein at 30 minutes postbypass, and CKBB at 6 hours postbypass. Piglets were killed 6 hours postbypass and their brains were harvested for histological/immunologic studies. Extent of damage was assessed using a semiquantitative score of 0 to 4 based on a validated method. RESULTS: Evidence for significant apoptosis and necrosis was apparent in all groups. The mean H&E score was 2.2 for group 1, 2.3 for group 2, 2.5 for group 3, and 2.3 for group 4. The mean terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling score was 1.0 for group 1, 1.2 for group 2, 1.7 for group 3, and 0.8 for group 4. Pathological changes were not greater in the piglets that did not have antegrade cerebral perfusion. Serum lactate, pyruvate, S-100B protein, and CKBB did not distinguish between perfusion strategies. CONCLUSIONS: In neonates, unmodified antegrade cerebral perfusion at flow rates of 10, 25, and 50 mL/kg per minute during DHCA does not provide additional protection of the brain as determined by histology, immunology, serum lactate, pyruvate, S-100B protein, and CKBB. PMID- 15793728 TI - Definitive surgical management of antenatally diagnosed exomphalos. AB - BACKGROUND/PURPOSE: The management of exomphalos is controversial with many centers in the United Kingdom and elsewhere advocating a conservative nonsurgical approach for the larger examples. Nevertheless, this approach is not without problems or complication. The aim of the study was to ascertain the outcome of all infants with an antenatally diagnosed exomphalos treated recently at our institution using a policy of aggressive abdominal wall closure. METHODS: This is a retrospective review of all infants with exomphalos treated from January 1995 to September 2002. RESULTS: There were 35 infants, all of whom underwent surgery. These were separated into 3 groups: group A (all exomphalos minor) underwent primary closure (n = 11), group B (exomphalos major) underwent primary closure (n = 13), and group C (exomphalos major) underwent staged closure involving a silo (n = 11). Infants in group C had a lower birth weight (P = .05) and were less mature (P = .06). They required longer periods of ventilation (P < .001), a longer hospital stay (P = .001), and a longer period to achieve full enteral feeds (P < .001). Overall survival was 34 (97%) of 35 infants. One premature infant who was born with a ruptured exomphalos sac (birth weight, 862 g) died of nonsurgical complications (sepsis and respiratory failure) early after the creation of a silo. CONCLUSIONS: An aggressive surgical approach in infants with exomphalos is a safe option resulting in effective abdominal wall closure. This requires a skilled multidisciplinary approach and possibly greater resources than other options. PMID- 15793729 TI - Gastroschisis: ward reduction compared with traditional reduction under general anesthesia. AB - BACKGROUND/PURPOSE: In gastroschisis it is proposed that gut reduction may be achieved without intubation or general anesthesia (GA) through ward reduction. The authors aimed to determine if ward reduction decreased morbidity and duration of treatment. METHODS: Infants born from January 1, 1995, to December 31, 2001, with gastroschisis were managed with either reduction under GA in the operating theatre (OT group)--up to September 1999, or ward reduction (when eligible) in the neonatal unit without GA/ventilation (ward reduction [WR] group)--from September 1999. RESULTS: Of the 37 infants, 31 were eligible for ward reduction 15 from the OT group, 16 from the WR group. All infants in the OT group had at least 1 episode of ventilation and 1 GA: 62% of infants in the WR group avoided ventilation (P = .0002) and 81% avoided GA (P < .0001). Infants who had ward reduction had significantly shorter durations of ventilation and oxygen therapy. Septicemia occurred in 31% of the WR group and 7% of the OT group (P = .17). Infants who had ward reduction left intensive care 16 days earlier (P = .02) and tended to reach full enteral feeds 8 days sooner (P = .06) and be discharged from hospital 15 days earlier (P = .05). CONCLUSIONS: Infants who had ward reduction do better in terms of avoiding GA/ventilation, establishing feeds, and going home earlier. A randomized, controlled trial comparing the 2 approaches is feasible, safe, and worthwhile. PMID- 15793731 TI - Increasing prevalence of gastroschisis in Utah. AB - BACKGROUND: Recent studies provide conflicting information about gastroschisis prevalence trends. The authors proposed that prevalence of gastroschisis in live births has increased in Utah and that characteristics of these infants would provide clinically useful information about treatment and outcomes. METHODS: Primary Children's Medical Center (PCMC) is the sole pediatric surgical referral hospital for Utah. The authors used both pediatric surgical and neonatal databases to identify gastroschisis cases at PCMC from 1971 through 2002. Only infants whose mothers had a primary residence in Utah were included. Individual charts were reviewed for infant characteristics for cases from 1998 through 2002. Utah Vital Statistics Reports were used to determine live birth rates and general infant and maternal characteristics. RESULTS: Gastroschisis prevalence increased from 0.36 to 3.92 cases per 10,000 live births over 31 years (P < .001). Young maternal age, primigravida status, and tobacco use were associated risk factors. Using the time required to achieve full enteric feedings at targeted volume and caloric density as a measurement of outcome, we found no association between delivery mode or surgical closure type (primary or secondary) and time to full feedings. Higher birth weight was associated with decreased time to full feedings (P = .03). CONCLUSIONS: Gastroschisis prevalence has increased 10-fold over the past 3 decades in Utah. PMID- 15793730 TI - Perinatal management of gastroschisis: analysis of a newly established clinical pathway. AB - PURPOSE: The authors developed a clinical pathway for optimal management after antenatal diagnosis of gastroschisis. This is the outcomes analysis of our first 30 consecutive patients. METHOD: Antenatal counseling was provided for all families with in-utero diagnosis of gastroschisis. Bowel dilatation, thickness, motility, amniotic fluid volume, and fetal development were followed by ultrasonography every 4 weeks. Babies were delivered by cesarean section between 36 and 38 weeks gestation if the lungs were mature or earlier for bowel complications. Gastroschisis repair was scheduled 90 minutes after birth. Primary repair was attempted in all through the abdominal wall defect without an additional incision, resulting in an umbilicus with no abdominal scar. RESULTS: Primary repair was achieved in 83%. Babies needed assisted ventilation for 3 days, reached full feeds by 19 days, and were discharged by 24 days (all medians). There were 3 (10%) deaths, all after staged repair. CONCLUSIONS: Our new protocol of both scheduled elective cesarean section and early gastroschisis repair resulted in a higher proportion of primary repair, shorter duration of mechanical ventilation, earlier full feeds, and shorter length of stay. There was no increase in mortality or morbidity. The primary-repair babies had no mortality and had excellent cosmesis. PMID- 15793732 TI - Immunoglobulin deposits in liver tissue from infants with biliary atresia and the correlation to cytomegalovirus infection. AB - PURPOSE: The aim of this report was to study the amount and distribution of immunoglobulin deposits in liver biopsies from infants with biliary atresia (BA) and correlate the results to the cytomegalovirus (CMV) infection status. METHODS: Stored liver biopsies from 18 patients with BA and from 6 control patients without liver disease were immunohistochemically stained to detect IgG and IgM deposits. The intensity of the immunoglobulin staining was evaluated by a semiquantitative scoring scale. Ongoing CMV infection was defined as the detection of CMV-IgM in serum and/or the isolation of CMV in the urine and was noted in 9 of the patients with BA. RESULTS: When analyzing the immunoglobulin deposits on the hepatocellular canalicular membrane the intensity score for IgM deposits was significantly higher in biopsies from patients with BA infected with CMV than in those without. No canalicular staining was detected in control biopsies. CONCLUSIONS: The results support the possibility that immunologic mechanisms are of importance in the pathogenesis of BA and that a CMV infection may trigger such mechanisms. PMID- 15793733 TI - Modified Soave procedure through the posterior sagittal approach for Hirschsprung's disease. AB - PURPOSE: The objective of this study is to describe the surgical technique and clinical results of a modified Soave procedure through the posterior sagittal approach (PSAP) for Hirschsprung's disease. METHODS: The patient was placed in a prone jackknife position. The rectum was approached via PSAP keeping the external sphincter intact. The rectum was isolated and then divided approximately 2 cm from the dentate line. The rectum and sigmoid were freed up to the normal segment. The aganglionic and ganglionic segments were confirmed intraoperatively by frozen biopsy. The rectal mucosectomy was started 0.5 to 1 cm proximal to the dentate line, leaving 1 cm of the muscular cuff. The colon was then pulled through the anus. The aganglionic segment and transitional zone were removed. Coloanal anastomosis was fashioned 0.5 to 1 cm proximal to the dentate line. RESULTS: Ninety-six patients were operated on from January 2000 to July 2002, using the described technique. Patient ages ranged from 2 to 36 months (median age, 8(1/2) months). The aganglionic segment was located in the rectum in 56 patients and in the sigmoid colon in 40 patients. The average length of bowel resected was 20 cm (range, 10-35 cm). Combined laparotomy was performed in 2 patients and combined laparoscopy surgery in 1 infant. There were no operative or postoperative deaths. Anastomotic leakage occurred in 2 patients. Spontaneous defecation occurred in all patients. CONCLUSIONS: Posterior sagittal approach provides excellent exposure of the operative field and decreases the major complications seen with the abdominal approach. The modified Soave procedure can easily be performed with PSAP. PMID- 15793734 TI - Fecoflowmetric profiles in postoperative patients with Hirschsprung's disease. AB - PURPOSE: The objective of this study is to evaluate the anorectal function from the viewpoint of fecoflowmetry in postoperative patients with Hirschsprung's disease (HD). METHODS: This study evaluated 23 long-term follow-up patients who had undergone a radical operation for HD. Their mean age was 11 years. The types of HD included rectosigmoid colon type, 18 cases, and entire colon type, 5 cases. An anorectal manometric study was performed before fecoflowmetry. After normal saline solution was administrated as an imitation stool into the rectal cavity under pressure monitoring, the patients defecated on a fecoflowmeter. After discussing the maximum defecation flow (Flow-max), fecoflow pattern (FFP), tolerance rate (TR), anal canal pressure (AP), and Kelly's clinical scores (Kelly Scores), the significant parameters were identified to elucidate the anorectal activity. RESULTS: (1) A close relationship was observed between the FFP and Kelly-Scores (P = .0027). (2) Flow-max, TR, and AP in patients with good Kelly Scores were significantly higher than those in patients with fair Kelly-Scores (P < .05). (3) The Flow-max accurately reflected the TR, Kelly-Scores, and AP. Flow max >45 mL per second, TR >70%, or AP >30 mm Hg was statistically regarded as a borderline level of fecal continence (P < .002). CONCLUSIONS: The Flow-max and FFP are considered to be useful parameters for postoperative patients with HD. PMID- 15793735 TI - Long-term outcomes of individualized biofeedback training based on the underlying dysfunction for patients with imperforate anus. AB - BACKGROUND: The aim of this study was to design an individualized biofeedback program based on the underlying dysfunctions and to assess the short-term and long-term clinical and quality-of-life outcomes for patients with imperforate anus. METHODS: Thirty-one patients were treated using an individualized biofeedback program according to their different dysfunctions after having completed the assessment of the anorectal function. Many special biofeedback training methods that consist of strengthening the perianal muscles biofeedback, sensory discrimination biofeedback, synchronizing external sphincter contraction biofeedback, and improving defecation dynamic biofeedback were selected to make up the individualized biofeedback training program. The clinical scores and quality-of-life scores were assessed in the short-term and long-term follow-up. RESULTS: The clinical scores and quality-of-life scores of patients before training, after biofeedback training, at short-term follow-up, and at long-term follow-up were 3.2 +/- 1.1 and 8.7 +/- 1.1, 5.2 +/- 0.6 and 11.0 +/- 0.8, 5.1 +/- 0.6 and 10.8 +/- 0.9, and 4.6 +/- 0.7 and 10.1 +/- 0.7, respectively. After biofeedback training, the strength of the perianal muscles were increased significantly, the abnormal rectal threshold sensation and defecation dynamic reverted to normal in all patients, and 82% patients who have no external anal sphincter reflex acquired the new reflex. In the short-term follow-up, 12 (57%) patients maintained the clinical outcome, and only 9 (43%) patients have regressions slightly. At long-term follow-up, 7 (33%) patients maintained the clinical outcome very well, and 14 (67%) patients had regressed. Although the clinical and quality-of-life scores have decreased slightly at follow-up, they were still significantly increased compared with those before biofeedback training. CONCLUSIONS: The individualized biofeedback is more suitable for improvement of the clinical outcome and the quality of life, and maintained a good clinical outcome and quality of life on the short-term and long-term follow ups. PMID- 15793736 TI - Laparoscopic management of Meckel's diverticulum in children. AB - BACKGROUND: Rectal bleeding, recurrent abdominal pain, nausea, and vomiting in children could present a diagnostic as well as therapeutic challenge. Meckel's diverticulum (MD) is one of the causes. The objective of the current study was to evaluate the feasibility and outcome of laparoscopic management of MD. METHODS: The clinical data of 33 children admitted with rectal bleeding and/or recurrent abdominal pain with no identifiable cause were reviewed over a period of 8 years. There were 23 boys and 10 girls with a mean age of 5.12 +/- 2 years (range, 3-12 years). In 21 cases, MD was an incidental finding on laparoscopic appendectomy and symptomatic in 12 cases. Patients with rectal bleeding were subjected to upper gastrointestinal endoscopy; colonoscopy, and technetium Tc 99m-labeled pertechnetate scan (MS). All patients were subjected to routine laboratory investigations and diagnostic laparoscopy. RESULTS: Of the 1200 appendectomies, incidental MD was found in 21 (1.9%) patients and symptomatic in 12 cases. Upper gastrointestinal endoscopy and colonoscopy did not show a bleeding source in 7 patients presented with bleeding per rectum. Four cases showed a positive MS uptake. Of these, 3 were found on laparoscopy to have an MD. Three cases showed a negative scan. Of these, 2 had an MD. In 5 cases with recurrent abdominal pain nausea, vomiting, and abdominal distention, diagnostic laparoscopy revealed Meckel's diverticulitis in 3 cases and intussusception secondary to MD in 1 case. Laparoscopic Meckel's diverticulectomy and laparoscopic-assisted Meckel's diverticulectomy was done for 18 and 12 cases, respectively. Ectopic gastric mucosa was present in 13 cases (44%). CONCLUSIONS: Laparoscopy is safe, cost effective, and efficient for the diagnosis and definitive treatment of MD. Compared with conventional laparotomy, it has the advantage of precise operative diagnosis, less traumatic access, fewer intraoperative and postoperative complications, and shorter recovery period. PMID- 15793737 TI - Are prophylactic antibiotics necessary with clean intermittent catheterization? A randomized controlled trial. AB - BACKGROUND/PURPOSE: Clean intermittent catheterization has been an established practice for more than 3 decades. The validity of antibiotic prophylaxis has been questioned although not tested. METHODS: Eighty-five patients were recruited into a randomized controlled trial. The randomization involved the placement into 1 of 2 groups: (A) continuing antibiotics or (B) discontinuing antibiotics. The trial would last 4 months with the outcome being a confirmed urinary tract infection. All groups were matched for age, sex, and pathology. RESULTS: The incidence of urinary tract infections was significantly increased in the group who continued to use antibiotics (n = 20) when compared with the group who discontinued prophylaxis (n = 3). The common infecting organism was Escherichia coli. CONCLUSIONS: The use of prophylactic antibiotics for children who intermittently catheterize may not be necessary. The use of prophylactic antibiotics may result in increased rates of infection because of the development of resistant organisms. PMID- 15793738 TI - Neonatal gastrointestinal stromal tumor. Report of a case and review of literature. AB - Gastrointestinal stromal tumor (GIST) is very rare in infancy. Most of the reported cases in the literature are in adults; some are in children but there are a few reported cases in the literature. The present case is a 6-day-old female neonate presenting with lethargy, poor feeding, constipation, abdominal distention, and rectal bleeding. She was operated on with the impression of intestinal obstruction, and right hemicolectomy was performed on her. Surgical specimen showed a well-defined and round 3-cm mass in the cecal area. Diagnosis was made by histologic and immunohistochemical studies which showed a GIST. The tumor showed positive vimentin and c-kit but negative for all other markers (desmin, actin, S100, NSE, and CD-34). So the case was an undifferentiated GIST. After 1 year of follow-up the patient was completely normal. PMID- 15793739 TI - The technique of laparoscopic retroperitoneal splenopexy for symptomatic wandering spleen in childhood. AB - BACKGROUND: Wandering spleen is an uncommon diagnosis, difficult to prove by standard investigations. The authors report a new method for laparoscopic splenopexy in children using a balloon-dilated retroperitoneal pouch. METHODS: From 3 accesses, the spleen is mobilized and displaced into a retroperitoneal pouch dilated to the double splenic volume. The pouch is dilated by a self-made balloon via a further intercostal access and narrowed by sutures incorporating the cranial and caudal edge of the gastrosplenic ligament. RESULTS: The peritoneal pouch contracts around the retroperitoneal spleen resulting in a firm fixation of the organ. This technique was successful in a 9-year-old girl with a 5-year history of severe recurrent abdominal pain. CONCLUSIONS: Laparoscopic retroperitoneal pouch splenopexy is a safe and effective procedure for symptomatic wandering spleen precluding the use of foreign materials in this age group. PMID- 15793740 TI - A case of neonatal neuroblastoma mimicking Altman type III sacrococcygeal teratoma. AB - The authors report a case of neonatal neuroblastoma mimicking Altman type III sacrococcygeal teratoma. A newborn male infant delivered after a normal pregnancy was found to have an extremely large sacrococcygeal mass. Imaging studies strongly suggested Altman type III sacrococcygeal teratoma. On the 10th day after the birth, the tumor was incompletely removed at surgery. Histopathologic examination of the tumor showed neuroblastoma. The tumor disappeared completely after chemotherapy. One year after diagnosis, no local recurrence or metastasis had been detected. To the authors' knowledge, this is the first case of neonatal neuroblastoma mimicking Altman type III sacrococcygeal teratoma. PMID- 15793741 TI - A case of incarcerated femoral hernia in an infant. AB - The authors describe the case of an incarcerated femoral hernia in an infant. A 4 month-old female infant was hospitalized with an incarcerated right groin hernia. Attempts to reduce the mass were unsuccessful, so an emergency surgery was performed. During surgery, the hernial sac seemed to protrude under the inguinal ligament. The authors opened the hernial sac and observed a dark red small intestine. The authors did not resect the intestine because its color gradually improved after the constriction was released. The hernial sac passed under the inguinal ligament, and the infant was diagnosed with incarcerated femoral hernia. PMID- 15793742 TI - Paravesical suture granuloma simulating a local recurrence of the immature sacrococcygeal teratoma. AB - It has been reported that paravesical suture granuloma after inguinal hernia repair may simulate a bladder, pelvic, or urachal malignancy in adults, and therefore, these lesions often pose a diagnostic and therapeutic dilemma. However, such a case has not been reported in the pediatric age group to date. The authors report a paravesical suture granuloma after left inguinal hernia repair in a child that mimics a local recurrence of the totally excised immature sacrococcygeal teratoma. PMID- 15793743 TI - When is a pneumothorax not a pneumothorax? AB - The authors report on a 13-year-old boy who, after exercise, had respiratory distress and left upper quadrant abdominal pain. Initially, a mistaken diagnosis of pneumothorax was made, and a chest tube was inserted. A nasogastric tube was then visualized on chest x-ray in the left hemithorax. He underwent a laparotomy and had herniation of spleen, stomach, and large and small bowel in the left pleural space passing through a traumatic defect in the hemidiaphragm. The laparoscopic Nissen fundoplication 3 years prior was felt to have contributed. A timely and correct diagnosis is essential to avoid the sequelae associated with these injuries and with inappropriate tube thoracostomy. PMID- 15793744 TI - Life-threatening airway obstruction caused by mediastinal germinoma in a 9-year old girl. AB - The authors report a case of a 9-year-old girl with a mediastinal germinoma that required emergent surgical extirpation because of tracheal compression and asphyxia. The tumor was successfully debulked under general anesthesia with the capability of extracorporeal circulation immediately available. Postoperatively, the tumor responded to adjuvant chemotherapy and irradiation and the patient survived without evidence of recurrence. PMID- 15793745 TI - Isolated cervical esophageal duplication: a rarity. AB - Duplication of alimentary canal is a well-known entity seen commonly in the abdomen and the thorax. Esophageal duplications are the second most common ones after the ideal duplications, but those in the neck are rarer-only 8 cases have been seen earlier. Cervical esophageal duplication in its isolated form causing torticollis and other chronic pressure symptoms has not been reported previously. Here, the authors present a 3-year-old male patient who had cystic duplication of the esophagus in the neck associated with the cervical vertebral defect and Horner syndrome. The authors hereby discuss the various embryological theories of etiopathogenesis of this malformation in brief. PMID- 15793746 TI - Cervical spondylodiscitis from an ingested pin: a case report. AB - In the pediatric literature, only 1 case of cervical spondylodiscitis from an ingested foreign body is reported and this was caused by a blunt radiolucent foreign body. The authors now describe a unique case of a 13-year-old teenaged boy who presented with neck pain 6 days after accidental ingestion of a sewing pin. Uncomplicated removal of this pin was followed in 36 days by the development of cervical spondylodiscitis that failed conservative management and required surgical debridement and arthrodesis. Physicians should be aware of the possibility of this complication in any patient that presents with neck pain after foreign body ingestion. PMID- 15793747 TI - Inflammatory pseudotumor of the ureter. AB - Inflammatory myofibroblastic pseudotumors are rare solid tumors found in most soft tissue locations although mainly in the lung. Their etiology is uncertain, and they are generally considered benign although some have a potential for recurrence and dissemination. Recent studies have suggested, however, that some of these tumors are in fact neoplastic processes that harbor chromosomal aberrations similar to those seen in certain lymphomas. The authors report a case of inflammatory pseudotumor of the ureter in a child and discuss recent reports. PMID- 15793768 TI - [ATM gene mutations in Chinese patients with ataxia telangiectasia]. AB - OBJECTIVE: To investigate the mutation characteristics of ATM gene in Chinese patients with ataxia-telangiectasia (AT). METHODS: Mutation of ATM gene was screened by polymerase chain reaction, reverse transcription-polymerase chain reaction, polyacrylamide gel electrophoresis combined with DNA direct sequencing in two Chinese AT patients. RESULTS: A missense mutation of 1346(G>C) in exon 11, which was a homozygotic mutation, was identified in one patient; a nonsense mutation of 610 (G>T) in exon 6 combined with a missense mutation of 6679 (C>T) in exon 47, which was a compound heterozygotic mutation, were identified in the other patient. They were co-segregated with the disease and were localized within the functional domain of ATM gene. CONCLUSION: Totally three novel ATM gene mutations were identified in two Chinese AT patients. PMID- 15793769 TI - [Mutations analysis in a pedigree with maternally inherited sensorineural hearing loss]. AB - OBJECTIVE: To analyze the mutations in a pedigree with maternally inherited sensorineural hearing loss, and to investigate whether 235delC heterozygote mutation in gap junction protein beta 2 (GJB2) gene modulates the severity of hearing loss associated with the A1555G mitochondrial mutation. METHODS: The PCR products were digested with the Alw26 I restriction enzyme, followed by direct sequencing to detect the mitochondrial mutations in 72 members of a core pedigree of an extensive family with matrilineal nonsyndromic deafness; 235delC mutation of the GJB2 gene was screened in this family by using the Apa I restriction enzyme and direct sequencing. RESULTS: The A1555G mutation of the mitochondrial DNA was present in all 27 members of maternal line, out of them, 21 members had phenotype of deafness (77.8%), with a high penetrance. Only three maternal line members of 72 members possessed 235delC heterozygote mutations, and the three had different phenotypes. CONCLUSION: The A1555G homozygous mutation of mitochondrial DNA is the susceptive etiological factor of nonsyndromic deafness in this family, but in the study of this pedigree, the 235delC heterozygous mutation in GJB2 gene may not aggravate the symptoms of hearing loss associated with the A1555G mitochondrial mutation. PMID- 15793770 TI - [Identification of a novel B variant allele in a Chinese Han individual with B subgroup]. AB - OBJECTIVE: To study the molecular genetic background of B subtype in Chinese Han population and identify novel allele at the ABO locus. METHODS: Ten samples from randomly selected blood donors of normal B phenotype used as control, and six samples from individuals diagnosed as B subgroup by serological tests were genotyped by sequence specific primer polymerase chain reaction and direct DNA sequencing at the exons 6 and 7 of ABO gene. The exons 6 and 7 and the intervening intron 6 of the B allele from each B subgroup sample were analyzed by cloning and haplotype sequencing. RESULTS: A novel B variant allele was identified in two individuals whose blood samples were diagnosed as belonging to Bx subgroup and Bw subgroup respectively, the novel B allele being different from the allele B101 by single 695T>C missense mutation in exon 7. A family with the individual possessed Bx subgroup was studied. Among 22 family members tested, seven family members were found to carry the novel B variant allele. No novel point mutation at the exons 6 and 7 of ABO gene were detected in the ten control samples and the other four samples with B subgroup. CONCLUSION: The present authors define this allele as a novel B variant allele. The mutation of this novel allele, in which the nucleotide alters from T to C at position of 695 in exon 7 and hence results in an amino acid change from Leu to Pro, is expected to diminish the enzyme's activity. It indicates that the alteration of amino acid at the position of 232 is critical to the activity of glycosyltransferases. PMID- 15793771 TI - [Identification of novel mutations in the phenylalanine hydroxylase gene of classical phenylketonuria]. AB - OBJECTIVE: To investigate the mutations of phenylalanine hydroxylase (PAH) gene in the cases of classical phenylketonuria (PKU). METHODS: The exons 3-12 of the PAH gene in 32 PKU patients from Inner Mongolia were studied by using PCR-single strand conformation polymorphism technique and DNA direct sequencing. RESULTS: Fourteen point mutations were identified. The frequencies of mutations were R243Q (12/64), Y356X (6/64), Y204C (5/64), R261Q (2/64), Y161S (2/64), R252Q (1/64), R111X (2/64), D282G (1/64), S303P (1/64), G239D (1/64), R413P (1/64), IVS7nt+2 (2/64), IVS4nt+3 (1/64) and IVS9nt+34 (2/64). Two novel mutations IVS4nt+3 (G>C) and IVS9nt+34 (G>A) were first found. The S303P (T>C) and D282G (A>G) were first documented in Chinese PAH gene. CONCLUSION: This study demonstrated the variety of the mutation type PAH gene of PKU in Inner Mongolia population, and confirmed that R243Q, Y356X, Y204C were the hot spots of PAH gene mutation. PMID- 15793772 TI - [C721T mutation of the alpha 1,3 galactosyltransferase gene responsible for Bw subgroup]. AB - OBJECTIVE: To gain an insight into the molecular genetic basis of Bw subgroup of ABO blood group system. METHODS: Three Bw phenotypes were confirmed by standard serological techniques. The enhancer, promoter and exons 1-7 including flanking introns of ABO gene were amplified and directly sequenced after PCR amplified fragments being purified by gel. Exons 6 and 7 were also sequenced after pcDNA3.1 (-) vector transformation. The sequence specific primer-polymerase chain reaction was performed to confirm the mutations detected by sequencing in this study. RESULTS: Genotypes of three individuals were Bw/O by direct sequencing, there were G deletion heterozygous at position 261 and C/T heterozygous at position 721. A normal O allele was confirmed by cloning sequencing and 721 C>T mutation of the alpha 1, 3 galactosyltransferase (B allele) gene was also observed, which caused amino acid 241 Arg>Trp substitution. This mutation was not detected in 140 random samples by PCR-SSP. CONCLUSION: The mutation of 721C>T in the alpha 1, 3 galactosyltransferase gene may be one of the molecular genetic bases of Bw phenotype. PMID- 15793773 TI - [Construction of DNMT1 siRNA stable expressing vector and evaluation of its silenced efficiency in blocking gene expression]. AB - OBJECTIVE: To construct the specific stable expression and high efficiency small interfering RNA(siRNA) expression vector that can block DNMT1 gene function. METHODS: Using vector-based RNA interference technique, the authors constructed a vector to transcribe functional short interfering RNA (RNAi). After transfection by lipofectmine (TM) reagent, the treated cells were selected by G418. The expression levels of RNA and protein of DNMT1 were analyzed by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting. The status of methylation of E-cadherin was analyzed by methylation-specific PCR(MSP). RESULTS: The expression level of endogenous DNMT1 mRNA in transfected SMMC-7721 cell lines with DNMT1 RNAi construct was 43% less than that in control cell 7721 pSU cell lines. The protein level in the former was about 10% less than that in the latter. The efficiency of the siRNA of DNMT1 was found to be higher than 90%. Demethylation of promoter of E-cadherin was obtained due to the inhibition of DNMT1. CONCLUSION: DNMT1 siRNA stable expressing vector was obtained by gene recombined technology. There was no complete sameness between the levels of protein and RNA in gene silenced cell lines. The efficiency of the siRNA should be confirmed by Western-blotting. PMID- 15793774 TI - [Construction and expression of siRNA expression constructs targeting epidermal growth factor receptor]. AB - OBJECTIVE: To construct the small interfering RNA (siRNA) expression constructs targeting epidermal growth factor receptor(EGFR) and express them in TJ905 human malignant cells. METHODS: Two target sequences from Receptor L domain and catalytic domain were selected to create two expression constructs using psiRNA NeoG2. Furthermore, the siRNA constructs were transfected into TJ905 cells as mediated by Lipofectamin. Meanwhile, an antisense EGFR construct p-anti-hEGFR was set as control. Immunofluorescence and Western blot were performed to detect EGFR expression. RESULTS: With the successful construction of the two siRNA expression plasmids and the stable transfection to TJ905 cells, the expression of EGFR was down-regulated to 90% and 92% respectively, but to 82% in the anti-sense EGFR group. CONCLUSION: The siRNA expression constructs targeting EGFR could specifically inhibit EGFR expression, and should be a new strategy in glioma gene therapy targeting EGFR. PMID- 15793775 TI - [The association of S447X and Hind III polymorphism in the lipoprotein lipase gene with dyslipidemia of the metabolic syndrome in patients with essential hypertension]. AB - OBJECTIVE: To assess the association of S447X mutation and Hind III polymorphism in the lipoprotein lipase gene with dyslipidemia of the metabolic syndrome in patients with essential hypertension. METHODS: A total of 983 patients were randomly selected from those with hypertension (diagnosed in the Community-based Comprehensive Studies on Prevention and Control of Hypertension Project in China) and those not treated with anti-hypertensive medications for at least in 2 weeks immediately before blood collection. Among them were 389 subjects with dyslipidemia and 594 subjects without dyslipidemia. The definition of dyslipidemia in patients with hypertension was used only when triglyceride or HDL cholesterol was at abnormal level. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to determine Ser447stop mutation and Hind III polymorphism in LPL gene. RESULTS: Linkage disequilibrium between the two sites was observed, with three major haplotypes identified: H+S, H-S, and H-X. The LPL gene S447X mutation and H-X haplotype were significantly associated with dyslipidemia (OR=0.547, 95%CI: 0.348-0.859 for S447X mutation; OR=0.537, 95%CI: 0.328-0.880 for H-X haplotype) in male, both by themselves and after adjustment for age, body mass index, smoking, alcohol intake, systolic blood pressure, diastolic blood pressure, education and serum glucose. The LPL H- carriers and H S haplotype were significantly associated with dyslipidemia (OR=0.575, 95%CI: 0.358-0.923) in female after multivariate adjustment. Moreover, compared with the H+S haplotype, the H-X haplotypes were associated with significantly lower TG and Log (TG/HDL-C) levels in both men and women, and with higher HDL-C levels in women; whereas no significant difference was observed between the H-S and H+S haplotype. Compared with the H-S haplotype, the H-X haplotypes had significant effect on the HDL-C levels in women. CONCLUSION: The LPL H-X haplotype was one of the protective factors of dyslipidemia of metabolic syndrome in hypertensive patients. It is significantly associated with low triglyceride, log triglyceride to-HDL-cholesterol ratio and high HDL-cholesterol levels. S447X mutation does not explain all the effect associated with the Hind III polymorphism, although the effect on serum lipids associated with the H-X haplotype appeared to be mainly mediated by the S447X mutation. It is possible that some functional mutations in the LPL gene besides the S447X mutation are in linkage disequilibrium with the Hind III polymorphism. PMID- 15793776 TI - [In silicon cloning of HV126, a novel human gene related to multi drug resistance in leukemia]. AB - OBJECTIVE: To find the novel gene related to the multi-drug resistance in leukemia and explore the molecular mechanism of multi-drug resistance. METHODS: The subtracted HL-60/VCR cDNA library was generated through the suppression subtractive hybridization using the wild HL-60 cells' cDNA as target and HL-60/ ATRA cells' as driver. A novel expression sequence tag (EST) sequence, which differentially expressed in HL-60/ ATRA cell, was screened by cDNA chip. Then a novel human gene, HV126 was assembled by the EST assembly tools. Bioinformatical databases and softwares were used to analyze and predict its function. Reverse transcription-PCR (RT-PCR) was used to detect the expression of HV-126 gene in leukemia cells before and after chemotherapy. RESULTS: The full open reading frames (ORFs) of the novel EST assembled by overlapping dbEST sequences included a 1991 bp nucleic sequence, which was named HV126. The deduced amino acid sequence consisted of 365 amino acids. The sequence of the novel gene exhibited 43% homology to a known gene, which is a possible member of the death domain flood family implicated in apoptosis and inflammation. The expression of HV126 was proved to be related to the drug sensitivity in leukemia cells by RT-PCR. CONCLUSION: HV126, the novel gene, might have roles in regulating multi-drug resistance in leukemia. Further laboratory research should be done on cloning and making clear the gene function. PMID- 15793778 TI - [Genetic polymorphisms of fifteen short tandem repeat loci in Chengdu Han population]. AB - OBJECTIVE: To acquire the population genetic data of fifteen short tandem repeat (STR) loci in Chengdu Han population. METHODS: A total of 210 EDTA-blood specimens were collected from the unrelated individuals in Chengdu Han population. The DNA samples were extracted with Chelex method and amplified by multiplex PCR technique. The PCR products were analyzed by an automatic genetic analyzer; the relative fragment's lengths of PCR products were calculated by gene scan analysis software and afterward genotyped by genotype software. RESULTS: Fifteen STR loci of the 210 samples showed a successful result of genotyping. The heterozygosities of the fifteen STR loci in Chengdu Han population were found to be 0.529-0.881; the combined exclusion probability and discrimination power for the fifteen STR loci in Chengdu Han population were determined to be 0.999998 and 7.3 x 10 (-17); respectively. CONCLUSION: The distinct genotype of fifteen STR loci and the sex of sample could be unveiled just through PCR and electrophoresis once, and a higher measured value could be obtained for both the combined discrimination power and the exclusion probability; the fifteen STR loci can meet the needs of the parentage testing and personal identification in forensic medicine. PMID- 15793777 TI - [The apolipoprotein E-CI-CII gene cluster polymorphisms and coronary artery disease]. AB - OBJECTIVE: To investigate associations between the apolipoprotein E-CI-CII gene cluster polymorphisms and coronary artery disease (CAD). METHODS: apoE genotypes were identified by multiplex amplification refractory mutation system (multi ARMS) and the polymorphisms of both apoCI and apoCII genes were detected by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 203 cases of CAD and 365 controls. Pairwise linkage disequilibrium coefficients (D, D') were estimated by the LINKAGE program. RESULTS: The frequencies of apoE E3/4 genotype (0.259) and epsilon4 (0.139) in CAD group were significantly higher than that in control group (0.125, 0.069), (P<0.05). The significant difference was also found for the apoCI locus, the frequencies of H2 allele were 0. 205 in the CAD and 0.113 in the control. Linkage disequilibrium coefficient D' was 0.672 (P<0.01) between apoE and apoCI genes. Significant differences for a deficit of epsilon3-H1-T1 and excess of epsilon4-H2-T1 were found in the CAD by estimation of the haplotype frequencies. After adjustment for possible confounding factors, the multivariate Logistic analysis showed a significant interaction among epsilon4, H2 and smoking, OR value was 18.3 (95%CI:2.35-150.81, P<0.05), attributable proportions of interaction (API) was 57.3%, it was a multiplicative model. An additive model was shown among epsilon4, H2 and bibulosity; the odds ratio (OR) (95%CI) and API of their interaction were 12.7(2.8-58.6, P<0.05) and 43.5%, respectively. CONCLUSION: The results suggested that both apoE and apoCI on chromosome 19 were the susceptibility loci for CAD, their linkage disequilibrium should be responsible for the development of CAD. Smoking and bibulosity can significantly increase the risk of CAD. PMID- 15793779 TI - [No evidence for genetic association between alpha-2 macroglobulin I1000V polymorphism and sporadic Alzheimer's disease in two independent Chinese populations]. AB - OBJECTIVE: Alpha-2 macroglobulin (alpha2M) is a proteinase inhibitor found in association with senile plaques in Alzheimer's disease (AD). Also alpha2M has been implicated in several pathophysiological processes in AD. In view of the recent contradictory reports on the relationship between AD and a common polymorphism I1000V in A2M gene, the present authors studied a relatively large sample, determined the genotype of the I1000V polymorphism in A2M gene in sporadic AD patients and age-matched controls with normal cognition, and examined the possible association of the polymorphism with AD. METHODS: Genotypes of A2M and apolipoprotein E (apoE) were detected by polymerase chain reaction combined with restriction fragment length polymorphism in 257 patients and 242 controls in Guangzhou, and 112 patients and 113 controls in Chengdu. RESULTS: The 1000Val allele frequencies in the merged AD and control groups were 7.7% and 8.7%, respectively. The differences of allelic and genotypic frequencies between the patients and control subjects were not statistically significant, even after stratification by apoE epsilon4 status or by age-of-onset of the disease. CONCLUSION: The results of this study revealed no association between the I1000V polymorphism of A2M and Chinese sporadic AD in Guangzhou and Chengdu. PMID- 15793780 TI - [Rapid detection of three common deletional alpha thalassemias in Chinese by single-tube multiplex PCR]. AB - OBJECTIVE: To develop a simple, rapid, accurate, and cost-effective single-0tube multiplex polymerase chain reaction (PCR) assay, which could be used for molecular screening and prenatal diagnosis, for detection of three commonest deletional alpha-thalassemias (-- (SEA), -alpha (3.7) and -alpha (4.2)) in Chinese population. METHODS: Four groups of primers were designed on the basis of gap-PCR, and the PCR reaction condition was optimized systematically with the purpose of amplifying effectively specific DNA fragments that are indicative of the respective genotypes of these three deletional alpha thalassemias. In addition, a pair of primers was designed to amplify LIS1 3' untranslated region (UTR) fragment for use as a separate control for amplification running. A total of 72 blood and prenatal archival DNA samples with various known alpha thalassemia genes or normal alpha globin gene sequence that had been confirmed by Southern blotting analysis or DNA sequencing were collected to test the specificity of this assay by blind analysis. In addition, DNA samples from nine couples at high risk of alpha thalassemia were also analyzed to evaluate the reliability of this technique in prenatal implementation. RESULTS: Homozygote, heterozygote and double heterozygote of the three commonest deletional alpha thalassemias were well detected simultaneously by this established method. For normal allele, a 2.4 kb amplified band as a systematic control and an alpha (2) gene-specific amplicon of 1.8 kb were produced. Besides the two amplified fragments of normal allele, it was found that a 1.3 kb, a 2.0 kb or a 1.6 kb amplified band could be simultaneously shown for representing --(SEA), -alpha (3.7) and -alpha (4.2) alleles, respectively, in the heterozygous states. In a blind test, this technique accurately detected 100% of the DNA samples previously characterized by Southern blotting or DNA sequencing, and it was successfully applied to prenatal diagnosis of alpha thalassemia in nine at-risk families. CONCLUSION: The single-tube multiplex PCR protocol presented in this study is easy-to-handle, rapid, reliable and is cost-effective for detecting --(SEA), alpha (3.7) and -alpha (4.2) chromosomes, and it is suitable for large-scale population screening and for rapid molecular genotyping in clinics. PMID- 15793781 TI - [Analysis of polymorphic markers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry]. AB - With the development of genomics and the accomplishments of human genomic sequencing, polymorphic markers and their analytic approaches are more and more important, and much attention has been paid to the fact that the analysis of single nucleotide polymorphisms utilizing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) is a high throughput approach. MALDI-TOF MS can also mini-sequence and genotype short tandem repeat. The approaches to analyzing single nucleotide polymorphisms are primer oligonucleotide base extension, ligase reaction, peptide nucleotide acid, invader assay, and so on. PMID- 15793782 TI - [Studies on PANK2 gene mutations in Chinese patients with Hallervorden-Spatz syndrome]. AB - OBJECTIVE: To study pantothenate kinase 2 (PANK2) gene mutations in Chinese patients with Hallervorden-Spatz syndrome (HSS). METHODS: PANK2 gene mutations were detected by PCR, DNA sequence analyses, restriction enzyme digestion and PCR single strand conformation polymorphism in 5 patients, 3 unaffected family members and 51 unrelated healthy persons. RESULTS: Novel compound heterozygous PANK2 gene mutations, A803G and T1172A, in exons 3 and 5, respectively, were found in one patient. At the same time, 3 types of single nucleotide polymorphisms, -38 t>a in 5'-UTR, IVS1+42 c>a and G77C in exon 1, were confirmed; among them, -38 t>a, IVS1+42 c>a, were first reported. CONCLUSION: PANK2 gene mutations can cause HSS in Chinese patients. PMID- 15793783 TI - [A recurrent rhodopsin gene missense mutation in a Chinese family with autosomal dominant retinitis pigmentosa]. AB - OBJECTIVE: To detect mutation in the rhodopsin gene (RHO) in a Chinese family with autosomal dominant retinitis pigmentosa (ADRP). METHODS: A total of 25 family members from a Chinese family were investigated. All the subjects were examined clinically by direct funduscopy, perimetry and vision test. Evaluation of the proband included electroretinography (ERG). Genomic DNA was extracted using standard method. The complete coding regions of RHO were amplified by polymerase chain reaction (PCR) and the PCR products were subjected to automatic DNA sequencing. RESULTS: 512 C>T (P171L), a recurrent missense mutation was detected in the proband. All 12 affected subjects in the family were heterozygous for the mutation. The affected individuals had night blindness at the age of 5-6 years. They had relatively severe impairment of visual acuity and suffered a gradual loss of peripheral visual field at the age of 20-30 years. And they went blind at the age of 40-50 years. Rod and cone ERG were not detectable in the proband. CONCLUSION: A recurrent missense mutation, 512C>T (P171L), was detected in a Chinese family with ADRP. PMID- 15793784 TI - [Screening 21-hydroxylase deficiency carriers in androgen excess women of Chinese Han nationality]. AB - OBJECTIVE: To gain a primary understanding of the prevalence of 21-hydroxylase deficiency(21-OHD) heterozygote (carrier) among androgen excess women of Chinese Han nationality, compare the molecular genetic changes therein revealed with the results of adrenocorticotropic hormone (ACTH) stimulating test, and assess the carriers' phenotype-genotype correlation. METHODS: Eighty-two androgen excess cases and 14 healthy women underwent ACTH stimulating test during the follicular phase. Molecular genetic analysis of CYP21 for 9 common mutations was performed with the method of amplification-created restriction sites. RESULTS: In androgen excess group, the basal level of F0 (P<0.01), as well as basal 17-OHP0 and the ACTH stimulated concentrations of 17-OHP60 were much higher than controls (P<0.01), and there was no obvious discrepancy in F60 (P>0.05). The net increase of 17-OHP and the ratio of net increase of 17-OHP to net increase of F were also higher than controls (P<0.01). No CYP21 gene mutations were found in control group. Four patients of the androgen excess group were identified as heterozygous carriers of CYP21 mutations. The ACTH stimulating test results from gene normal patients and from carriers overlapped to a certain extent. CONCLUSION: Among 82 patients of Chinese Han nationality androgen excess women, 4.9% were 21-OHD heterozygous. The response of serum 17-OHP is not useful for predicting CYP21 gene mutation carrier status. Genotyping is the most reliable method to detect carrier. PMID- 15793785 TI - [Mitochondrial DNA 3243, 3316 point mutations and type 2 diabetes mellitus]. AB - OBJECTIVE: To investigate the prevalence of mitochondrial DNA (mtDNA) mutation at position 3243(A/G) and 3316(G/A) in Chinese patients with type 2 diabetes mellitus. METHODS: Two hundred and twenty-five unrelated Chinese patients with type 2 diabetes and 195 nondiabetic control individuals without family history of diabetes in Yunnan were examined. The presence of mt 3243 and 3316 mutations was determined by polymerase chain reaction amplification and restriction fragment length polymorphism (PCR/RFLP). Finally, mutant mtDNA was confirmed by DNA sequencing. RESULTS: The mitochondrial DNA mutation at position 3316 was found in 5 of 225 (2.22%) patients with type 2 diabetes, and this mutation was found in 2 of 195 (1.03%) control individuals. There was no significant difference in respect to the frequency of the mutation between patients and controls. The mitochondrial DNA mutation at position 3243 was not found in any of the patients and controls. CONCLUSION: The results suggest that the prevalence of the mitochondrial tRNA (Leu(UUR))gene at position 3243(A/G) mutation is so low that it may not be a major cause of type 2 diabetes mellitus in patients of Yunnan, China, and the mitochondrial ND1 gene at position 3316(G/A) mutation may be a polymorphism unrelated to diabetes in Chinese. The role of other genetic, environmental and intrauterine factors needs further investigation. PMID- 15793786 TI - [Inherited dysfibrinogenemia caused by Arg275His in the beta chain of fibrinogen]. AB - OBJECTIVE: To analyze the phenotype and genotype of a family with inherited dysfibrinogenemia. METHODS: Laboratory tests including activated particle thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), and the activity of protein C (PC), protein S(PS) and antithrombin (AT) were conducted in the proband and 4 family members. The activity and antigen of fibrinogen in plasma were measured by functional and immunoturbidimetry assay, respectively. All the exons and exon-intron boundaries of the three fibrinogen genes were analyzed by direct sequencing. RESULTS: The proband had normal APTT and PT, but prolonged TT. Her plasma fibrinogen levels were extremely reduced, which was also found in her mother. The sequencing results of the proband revealed heterozygous g.5678 G>A in the exon 8 of FGG gene originating from her mother, which caused Arg275His missense mutation. CONCLUSION: Dysfibrinogenemia in the family is caused by Arg275His in the beta chain of fibrinogen and it is the first report on a Chinese family with inherited dysfibrinogenemia. PMID- 15793787 TI - [Influence of angiotensin-converting enzyme gene polymorphism on the anti proteinuria efficacy of angiotensin-converting enzyme inhibitor in Han nationality of southern Sichuan province in China]. AB - OBJECTIVE: To investigate the association between angiotensin-converting enzyme (ACE) gene polymorphism and the reducing urinary protein efficacy of angiotensin converting enzyme inhibitor (ACEI) in patients with primary chronic glomerulonephritis in Han nationality of southern Sichuan province. METHODS: Ninety-nine primary glomerulonephritis patients with urinary protein were enrolled in this study. They were treated with benazepril for at least 3 months. The ACE gene insertion/deletion(I/D) polymorphisms in intron 16 were determined by PCR. A comparison of the reducing urinary protein efficacy of benazepril was made between the patients with different ACE genotypes. RESULTS: Urinary protein excretion was significantly higher in patients with ACE DD genotype than that in patients with II genotype. After benazepril treatment for 3 months, the rates of urinary protein decline were observed. The rates of reduction of proteinuria in patients with DD genotype and ID genotype were obviously higher than that in patients with II genotype(P<0.05). CONCLUSION: Benazepril could decline the rate of urinary protein excretion in patients with primary chronic glomerulonephritis, and significant relationship was observed between ACE gene polymorphism and the reducing urinary protein efficacy of ACEI. PMID- 15793788 TI - [Angiotensin converting enzyme gene and exercise-induced silent myocardial ischemia in type 2 diabetes mellitus]. AB - OBJECTIVE: To investigate the relationship between angiotensin converting enzyme (ACE) gene and exercise-induced silent myocardial ischemia (SI) in patients with type 2 diabetes mellitus. METHODS: One hundred and eight patients suffering from type 2 diabetes mellitus with normal rest electrocardiograph and 50 healthy individuals were selected randomly. SI was diagnosed with treadmill exercise test and ACE genotypes were detected with PCR. RESULTS: (1) The control group and type 2 diabetes mellitus group had similar distribution of ACE genotypes and alleles (P>0.05). Compared with the non-SI group, the SI group had significantly higher ACE D allele prevalence (Chi-square=4.501, P<0.05); however, the two groups had similar prevalence of ACE genotypes (P>0.05). (2) There were no significant differences in clinical characteristics and serum lipoproteins among the three ACE genotypes (II, DD,ID) of type 2 diabetes mellitus (P>0.05). (3) The prevalence of SI in DD group was found to be 68.2%, which was significantly higher than that in II genotype group (39.5%, Chi-square=4.593, P<0.05). CONCLUSION: ACE D allele increases the risk of SI in type 2 diabetes mellitus. PMID- 15793789 TI - [Rapid prenatal detection of Down syndrome by homologous gene quantitative PCR]. AB - OBJECTIVE: To evaluate the use of homologous gene quantitative PCR (HGQ-PCR) as a method for non-invasive diagnosis of Down syndrome and for prevention of the birth of Down syndrome children. METHODS: HGQ-PCR, which can directly detect the additional copy of chromosome 21 by comparing simultaneously amplified two highly homologous genes, i.e. the human liver-type phosphofructokinase located on chromosome 21 critical region of Down syndrome (PFKL-CH21) and the human muscle type phosphofructokinase located on chromosome 1 (PFKM-CH1), was performed in 38 clinically diagnosed Down syndrome patients and 178 normal controls. RESULTS: The ratios of PFKM-CH1/PFKL-CH21 products were 1.40 +/- 0.367 (mean +/- SD) and 0.46 +/- 0.21 (mean +/- SD) for disomy 21 and trisomy 21, respectively. The difference between these two groups was statistically significant (P<0.001). CONCLUSION: This approach has proven to be a practical and direct method for the detection of trisomy 21 and may also be applied to the detection of the extra piece of 21q involved in translocation-type of Down syndrome. PMID- 15793791 TI - [Relationship between the microsatellite polymorphism of CYP11 alpha gene and the pathogenesis of hyperandrogenism of polycystic ovary syndrome in Chinese]. AB - OBJECTIVE: To explore the association between the microsatellite polymorphism in the promoter region of CYP11 alpha gene and hyperandrogenism of polycystic ovary syndrome (PCOS). METHODS: Eighty-six cases of PCOS and 50 normal women as controls were studied. Polymerase chain reaction and electrophoresis on polyacrylamide gel were employed to detect the polymorphism of CYP11 alpha gene and its frequency distribution. At the same time, the relationships of CYP11 alpha alleles to serum testosterone levels in PCOS were compared. RESULTS: Four different CYP11 alpha (tttta)n alleles were identified, corresponding to 4, 6, 8 and 9 repeat-units alleles. The frequency distribution profiles were 0.17, 0.31, 0.39, 0.13 and 0.22, 0.35, 0.33, 0.10 in PCOS group and control group respectively, showing no statistically significant difference between the two groups. There were no correlations between the polymorphism of CYP11 alpha gene and the serum testosterone levels of PCOS patients. CONCLUSION: Micro-satellite polymorphism (tttta)n of gene CYP11 alpha exists in Chinese women and the polymorphism does not relate to the pathogenesis of hyperandrogenism in women with PCOS. PMID- 15793792 TI - [5, 10-methylenetetrahydrofolate reductase polymorphism in three nationalities of Guizhou in China]. AB - OBJECTIVE: To study the genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) among the Han, Buyi and Miao populations in Guizhou and to provide genetic data for establishment of the genetic polymorphism bank of Guizhou Minorities. METHODS: The technique of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotype and allele frequencies at two mononucleotide sites (677 and 1298) of MTHFR among the Han population in Libo county, the Buyi population in Libo county and the Miao population in Leishan county. RESULTS: At the site of 677, the T allele frequencies were found to be 22.8%, 16.1%, 10.6%, for the Han, Buyi, Miao populations respectively. At the site of 1298, the C allele frequencies were 28.9%, 39.1%, 48.7% for the Han, Buyi, Miao populations respectively. The frequencies for the combined heterozygote of 677CT/1298AC were 16.66%, 22.7%, 11.1% for the three populations respectively. Moreover, one case with combined homozygote of 677TT/1298CC was seen in the Miao population. CONCLUSION: The polymorphisms of the two mononucleotide sites (677 and 1298) of MTHFR are diverse in different populations. The C allele frequencies at the site of MTHFR 1298 of the Miao population in Leishan county and the Buyi population in Libo county are high, and the C allele frequency in the Miao population is higher than those hitherto reported in literature. PMID- 15793790 TI - [The clinical significance of interphase fluorescence in situ hybridization monitoring chimeric status after sex-mismatched allogeneic hematopoietic stem cell transplantation for leukemia]. AB - OBJECTIVE: To explore the association between chimerism, minimal residual disease (MRD) and relapse after sex-mismatched allogeneic hematopoietic stem cell transplantation (allo-HSCT) for leukemia. METHODS: Fifty-seven patients with leukemia received allogeneic hematopoietic stem cell grafts from HLA-matched or partially matched, but sex-mismatched donors. Chimeric status and MRD were detected by dual-color interphase fluorescence in situ hybridization (I-FISH) using X/Y sex chromosome centromere DNA probe and bcr/abl dual fusion DNA probe, respectively, at different time points after transplantation. SPSS software was used to analyse the correlation between chimeric status, MRD and relapse. RESULTS: In comparison with karyotype analysis, I-FISH was of higher sensitivity in detecting sex chromosome and bcr/abl fusion gene. Chimeric status was negatively correlated with MRD (r=-0.9690, P<0.01). In the early times of transplantation (within 3 months), mixed chimerism had higher relapse rate than did complete chimerism. Chimeric status and MRD were correlated with leukemic relapse (r=-8240, P<0.01; r=-0.9040, P<0.01). The decrease in chimeric status occurred before leukemic relapse in hematology. CONCLUSION: I-FISH is a more specific and sensitive test for monitoring MRD after transplantation. The clinical value of sex chromosome is identical to that of the special tumor gene for monitoring MRD after transplantation. Chimeric status is negatively correlated with MRD. Chimeric status and MRD are associated with leukemic relapse. The decrease in chimeric status is considered a mark of leukemic relapse after transplantation. PMID- 15793793 TI - [Study of genetic polymorphism of 6 short tandem repeat loci in Nongqu Mongolian of inner Mongolia Autonomous Region in China]. AB - OBJECTIVE: To get the genotype and allele frequency distribution of 6 short tandem repeat (STR) loci VWA, FGA, PENTAE, D6S1043, D2S1772, D7S3048 in NongQu Mongolia of China. METHODS: Two hundred and ninety-three unrelated individuals from Nongqu Mongolian were investigated. Polymerase chain reaction and polyacrylamide gel electrophoresis were used. RESULTS: Eighty alleles and 335 genotypes were detected, with frequencies ranging from 0.0017 to 0.2828. All the 6 loci met Hardy-Weinberg equilibrium. The statistical analysis of 6 STR loci showed the heterozygosity (H) >/= 0.7945, the discrimination power (DP) >/= 0.9160, the probability of paternity exclusion (PPE) >/= 0.5919, and the polymorphic information content (PIC) >/= 0.7617. CONCLUSION: These results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic forensic medical application. PMID- 15793794 TI - [Distribution of apoE polymorphism in Chinese Yunnan Dehong Dai ethnic group]. AB - OBJECTIVE: To investigate the distribution of apolipoprotein E (apo E) polymorphism in Chinese Dehong Dai and Kunming Han ethnic groups. METHODS: After the process of extracting genomic DNA from 171 Chinese Dehong Dai and 71 Chinese Kunming Han subjects, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to amplify the fourth exon of apoE, which contains site 112 and 158 in amino acid sequence, and the apoE polymorphism was detected. RESULTS: The genotyping frequencies of apolipoproteinE epsilon 2/2, epsilon 2/3, epsilon 2/4, epsilon3/3, epsilon3/4, epsilon4/4 in Dehong Dai were 0.006, 0.111, 0.006, 0.789, 0.088 and 0.000 respectively, while the genotyping frequencies of apoE epsilon2/2, epsilon2/3, epsilon2/4, epsilon3/3, epsilon3/4, epsilon4/4 in Kunming Han were 0.000, 0.169, 0.014, 0.718, 0.099, 0.000, respectively (P>0.05). The allelic frequencies of apoE epsilon2, epsilon3 and4 in Dehong Dai were 0.064, 0.889, 0.047 respectively, while the allelic frequencies of apoE epsilon2, epsilon3 and epsilon4 in Kunming Han were 0.092, 0.852, 0.056, respectively (P>0.05). CONCLUSION: The results revealed that either the frequency of genotype or that of allele of apoE gene polymorphism showed statistical difference between the ethnic populations. Compared with the data from other nationalities in China, the frequency of epsilon2 allele in Dehong Dai was significantly lower than that in Zhuang nationality (P<0.01); the frequency of epsilon3 allele in Dehong Dai was significantly higher than that in Chaoxian, Hui, Mongolian,Zhuang nationalities (P<0.05) and Uygur nationality (P<0.01); the frequency of epsilon4 allele in Dehong Dai was significantly lower than that in Olunchun nationality (P<0.05) and Uygur, Owenk nationalities (P<0.01). By comparison with the data from the populations of other races, the distribution of apoE gene polymorphism in Dehong Dai population is similar to that in Japanese population (P>0.05), but different from those in Singapore,European and American populations. PMID- 15793795 TI - [Determination of HLA-A, -B allele polymorphism in the Luoba nationality living in Tibet Autonomous Region in China]. AB - OBJECTIVE: To investigate the HLA-A, -B allele polymorphism in the Luoba ethnic population. METHODS: HLA-A, -B DNA types in 92 healthy individuals of Luoba nationality in the Linzhi area, Tibet Autonomous Region, were investigated by polymerase chain reaction-sequence specific oligo-nucleotide (PCR-SSO). RESULTS: Ten alleles at HLA-A locus, and 19 alleles at HLA-B locus in Luoba ethnic group were detected. Of the 10 HLA-A alleles detected, the three most common alleles were HLA-A*11(allele frequency: 36.40%), -A*02 (25.50%), -A*24 (23.90%), and they covered 85.80% of the total HLA-A alleles detected from the Luoba ethnic group. Of the 19 HLA-B alleles detected, the three most common alleles were HLA-B*40 (27.20%), -B*15 (11.40%) and -B*38(10.90%), and they covered 49.50% of the total B alleles detected in the Luoba ethnic group. CONCLUSION: The distribution of HLA A, -B allele polymorphism in the Luoba nationality is distinctive, but some of the gene distribution in the Luoba group is nearer to that in the Tibetan group. These are consistent with the results of ethnological, historical and sociological researches. PMID- 15793796 TI - Optimization and characterization of a sphingomyelin/cholesterol liposome formulation of vinorelbine with promising antitumor activity. AB - Vinorelbine (VRL) is a particularly lipophilic member of the vinca alkaloids which, as a class of drugs, exhibit improved cytotoxicity and therapeutic activity through increased duration of exposure. Here, we describe and optimize a sphingomyelin/cholesterol (SM/Chol) liposome formulation of VRL to maximize in vivo drug retention, plasma circulation time, and therapeutic activity. VRL was efficiently encapsulated (>90%) into 100 nm liposomes using an ionophore-mediated loading method. VRL retention in SM/Chol liposomes after intravenous injection in mice was dependent on drug-to-lipid ratio (D/L), with higher D/L ratios exhibiting increased drug retention (0.3 > 0.2 > 0.1, wt/wt) and improved pharmacokinetics. Cryo-electron microscopic examination of a high D/L ratio formulation indicated that the intravesicular regions of these liposomes were electron dense compared with empty liposomes. The optimized, high D/L ratio SM/Chol VRL formulation showed promising activity against subcutaneous B16 melanoma tumors compared with VRL or SM/Chol formulations of vincristine or vinblastine. Finally, the stability of the formulation was excellent (<5% drug leakage, >99% intact VRL, no changes in liposome size after 1 year at 2-8 degrees C). The optimized drug retention properties of the SM/Chol formulation of VRL, combined with its promising antitumor activity and pharmaceutical stability, make this formulation an excellent candidate for future clinical development. PMID- 15793797 TI - A chimeric GB virus B with 5' nontranslated RNA sequence from hepatitis C virus causes hepatitis in tamarins. AB - Only humans and chimpanzees are fully permissive for replication of hepatitis C virus (HCV), an important cause of liver cirrhosis and cancer worldwide. The absence of suitable animal models limits opportunities for in vivo evaluation of candidate hepatitis C therapeutics and slows progress in the field. Here, we describe a chimeric virus derived from GB virus B (GBV-B), an unclassified hepatotropic member of the family Flaviviridae that is closely related to HCV and infects tamarins (Saguinus sp.), in which a functionally important HCV regulatory sequence replaced an analogous sequence in the 5' nontranslated region (5'NTR) of the GBV-B genome. The transplanted sequence comprised domain III of the internal ribosome entry site (IRES), which directly binds the 40S ribosome subunit and is a target for candidate therapeutics. The chimeric 5'NTR retained ribosome binding activity and was competent in directing protein translation both in cell-free translation reactions and in transfected primary tamarin hepatocyte cultures. Virus rescued from the chimeric RNA replicated in the liver of tamarins, causing biochemical and histopathological changes typical of viral hepatitis. However, adaptive mutations were required elsewhere in the genome for efficient replication. Virus was not rescued from other, translationally competent, chimeric RNAs in which domain II of the IRES was exchanged. Thus, the 5'NTR appears to contain virus-specific replication signals that interact with other sites within the viral genome or with viral proteins. In conclusion, such novel chimeric flaviviruses offer opportunities for new insights into HCV replication mechanisms, while potentially facilitating the evaluation of candidate therapeutics in vivo. PMID- 15793798 TI - Effect of particle shape of active pharmaceutical ingredients prepared by fluidized-bed jet-milling on cohesiveness. AB - Milling is a common procedure to improve bioavailability of many active pharmaceutical ingredients (APIs), which typically have low solubility in water. But such micronization can yield an increase in the cohesiveness of particles. Although particle cohesiveness is desirable for tablet strength in the subsequent formulation process, increased particle cohesiveness can lead to operational difficulties in a milling equipment due to compaction of particles inside. In this article, the impact of milling via a fluidized-bed jet-mill on the cohesive strength and interparticle force was studied using Ethenzamide as a pharmaceutical model compound. As a result, the particle shape was found to affect both the tensile strength of powder bed and the interparticle cohesive force. A powder bed, having relatively high void fraction by direct tensile test, shows a positive correlation between the cohesive force and the particle sphericity, while powders with low void fraction by diametral compression test show a positive correlation between the cohesive force and the angularity of the particle. PMID- 15793799 TI - A novel standard sample powder preparation method for quantitative analysis of polymorphs. AB - The purpose of this study was to improve the sensitivity and accuracy of the quantitative analysis of polymorphic mixtures. Calibration curve reliability has been improved through the advancement of standard mixture preparation techniques whereby samples at scales of several grams are homogeneously and accurately prepared without decrease in crystallinity. Crystalline powders are first micronized to anisotropic, nearly equal-sized (<10 microm) particles of almost random orientation in an air jet mill. These powders are then mixed in a vibration mill that includes rubber balls as mixing media. Using this technique, alpha and gamma forms of indomethacin (IMC) were prepared in various proportions from 0.5% to 50% alpha content and subjected to X-ray powder diffraction analysis. The validation data showed that these precisely prepared standard mixtures resulted in superior analytical sensitivity and accuracy compared to those prepared by agate mortar. PMID- 15793800 TI - Physicochemical characterization of ricobendazole: I. Solubility, lipophilicity, and ionization characteristics. AB - The physicochemical properties of ricobendazole (RBZ) were characterized using conventional methods. Solubility in some pharmaceutical solvents, pH-solubility, ionization properties, and lipophilicity are described. The solubility of RBZ in water is 62 mug/mL and very poor in common pharmaceutical solvents, for example, oils (<0.25 mg/mL in all the tested oils) or ethanol (1.2 mg/mL) and propylene glycol (2.6 mg/mL), and slightly higher in dipolar solvents, DMSO (16.5 mg/mL). U shaped pH-solubility profile in aqueous solutions indicates RBZ is an ampholyte. pK(a) values measured by absorbance spectroscopy and pH solubility methods were 3.45 and 3.76 for the basic group and 9.82 and 9.53 for the non-basic nitrogen, respectively. Combination of low pH and surfactant/co-solvent mixtures also improved solubility. RBZ formed a 1:1 complex with hydroxypropyl-beta cyclodextrin (HP-beta-CD) with a binding constant (K(1:1)) of 311/M1. Apparent partition coefficients of RBZ were 14.3-15.2 at pH 6-9 and reduced at higher or lower pH. In conclusion, traditional organic co-solvents, pH-adjustment or complexation (with HP-beta-CD) approaches are unlikely to yield sufficient solubility for formulation of RBZ solutions for subcutaneous injection and novel approaches should be considered. PMID- 15793801 TI - Solid films of blended poly(vinyl alcohol)/poly(vinyl pyrrolidone) for topical S nitrosoglutathione and nitric oxide release. AB - Nitric oxide (NO) is responsible for biological actions in mammals, ranging from the control of arterial pressure to immunological responses. In this study, S nitrosoglutathione (GSNO), a spontaneous NO donor, was incorporated in solid films of blended poly(vinyl alcohol) (PVA) and poly(vinyl pyrrolidone) (PVP) comprising a biomaterial with potential for the local delivery of NO. In dry conditions, the extinction of the absorption bands of GSNO was correlated with the increase of the absorption band of its dimmer, GS-SG, implying NO release through the homolytic cleavage of the S-N bond. Mass spectrometry was used to confirm and to monitor the release of free NO from solid PVA/PVP-GSNO films to the gas phase. Kinetic measurement based on the Griess reaction was used to show that solid PVA/PVP-GSNO films are also capable of releasing both NO and GSNO to aqueous solution trough diffusion. Storage experiments have shown that GSNO is highly stabilized in the dry PVA/PVP matrix. The results indicate that GSNO containing PVA/PVP films may be used for delivering free NO and/or GSNO topically and controllably. PMID- 15793802 TI - Oral heparin delivery: design and in vivo evaluation of a stomach-targeted mucoadhesive delivery system. AB - Low molecular weight heparin (LMWH) is an agent of choice in the anti-coagulant therapy and prophylaxis of thrombosis and coronary syndromes. However, the therapeutic use is partially limited due to a poor oral bioavailability. It was therefore the aim of this study to design and evaluate a highly efficient stomach targeted oral delivery system for LMWH. In order to appraise the influence of the molecular weight on the oral bioavailability, mini-tablets comprising 3 kDa (279 IU) and 6 kDa (300 IU) LMWH, respectively, were generated and tested in vivo in rats. The potential of the test formulations based on thiolated polycarbophil, was evaluated in comparison to hydroxyethylcellulose (HEC) as control carrier matrix. The plasma levels of LMWH after oral versus subcutaneous administration were determined in order to calculate the relative bioavailability. With the delivery system containing 3 kDa LMWH (279 IU) a relative bioavailability of 19.1% was achieved, offering a significantly (p < 0.05) better bioavailability than the control system displaying a relative bioavailability of 8.1% The 6 kDa LMWH (300 IU) formulation displayed a relative bioavailability of 10.7% in contrast to the control displaying a relative bioavailability of 2.1%. In conclusion, these results suggest that mucoadhesive thiolated polymers are a promising tool for the non-invasive stomach-targeted systemic delivery of LMWH as model for a hydrophilic macromolecular polysaccharide. PMID- 15793803 TI - Effect of piceatannol in human monocyte-derived dendritic cells in vitro. AB - Piceatannol is an anti-inflammatory, immunomodulatory, and antiproliferative stilbene that has been shown to interfere with the cytokine signaling pathway. Dendritic cells (DCs) play a pivotal in the initiation of T-cell-mediated immune responses, making them an attractive cellular adjuvant for use in cancer vaccines. This study investigated the effect of piceatannol on the phenotypic and functional maturation of human monocyte-derived DCs in vitro. Human monocytes were cultured with GM-CSF and IL-4 for 6 days, followed by another 2 days in the presence of piceatannol or LPS. DCs harvested on day 8 were examined using functional assays. The expression levels of CD1a, CD80, CD83, and CD86 as expressed by mean fluorescence intensity (MFI) on DCs differentiated from immature DCs after culture with 1 muM of piceatannol for 2 days were enhanced and decreased endocytic activity. Piceatannol-treated DCs also displayed enhanced T cell stimulatory capacity in a MLR, as measured by T-cell proliferation. Similar results were obtained with DCs differentiated with LPS from immature DCs. However, piceatannol did not inhibit phenotypic and functional maturation induced by LPS from immature DCs. Piceatannol-treated DCs induced the differentiation of naive T cells towards a helper T-cell type 1 (Th1) response at DCs/T (1:5) cells ratio depending on IL-12 secretion. These results demonstrate that piceatannol may be used on DC-based vaccine for cancer immunotherapy. PMID- 15793804 TI - Evaluation of indomethacin percutaneous absorption from nanostructured lipid carriers (NLC): in vitro and in vivo studies. AB - The aim of this study was the evaluation, in vitro and in vivo, of indomethacin (IND) release through the skin from nanostructured lipid carriers (NLC). NLC were prepared by ultrasonication, and were characterized in order to determine drug content, and particle size; finally the NLC were processed to hydrogels (A and B). The IND release pattern from NLC hydrogels was evaluated in vitro, to determine its percutaneous absorption through excised human skin (stratum corneum and epidermis, SCE), and in vivo. To evaluate the in vivo IND release, two methods were employed: (1) the IND topical anti-inflammatory activity was determined at different time-points after its cutaneous application; in this case, the UVB-induced erythema on healthy human volunteers, chosen as inflammatory model, was monitored by reflectance visible spectrophotometry; (2) the extent of IND absorption into human skin was performed by the tape-stripping technique. The in vitro percutaneous absorption studies showed lower fluxes of IND through SCE membranes from NLC hydrogels (A and B) in comparison to an aqueous dispersion (C) and a hydro-alcoholic gel (D) both containing free IND. The findings from the former in vivo method showed that the anti-inflammatory effect, following IND topical application, was more prolonged with IND-loaded NLC gel formulation (A) if compared to formulation C and D. The results from tape stripping technique confirmed the trend obtained by the former in vivo method and indicated that IND topical bioavailability in the stratum corneum varied substantially depending upon the formulations (A-D). PMID- 15793805 TI - Calorimetric investigation of the structural relaxation of amorphous materials: evaluating validity of the methodologies. AB - Although the potential advantages of the amorphous solid state is widely recognized among pharmaceutical researchers, its industrial applications have been mainly limited to freeze-dried injectable formulations where the amorphous form is naturally produced. Applications in oral dosage forms have been limited due, at least in part, to the poor state of knowledge regarding physical properties and stability of amorphous materials. Relaxation behavior is perhaps one of the most important physical characteristics of amorphous materials because relaxation kinetics are closely related to physical and chemical stability. Although recent developments in calorimetry methodology have facilitated detailed characterization of relaxation behavior, some experimental difficulties remain, and quantitative analysis of structural relaxation is still under development. This review focuses on the calorimetric investigation of the structural relaxation of drugs and excipients, and discusses the difficulties in the experimental evaluation of the relaxation time by those methods. We also present an original investigation of the impact of increases in relaxation time during an annealing experiment on the values of relaxation time, tau, and stretched exponential constant, beta, obtained from analysis of the experiment according to the Kohlraush-Williams-Watts kinetic model. Using results from a numerical simulation, we find that the values of tau and beta obtained from the data analysis are too large and too small, respectively, but the value of stretched relaxation time, tau(beta), remains reliable. The time dependence of the relaxation time is likely to play an important role in the non-Arrhenius behavior of pharmaceutical glasses. PMID- 15793807 TI - Immobilization of ibuprofen and copper-ibuprofen drugs on layered double hydroxides. AB - The immobilization of the NSAID ibuprofen (Hibp) and the Cu(II)-ibp compound on magnesium-aluminum layered double hydroxides (Mg3Al-LDH) is described. Ibuprofen was intercalated on LDHs by three routes (ion exchange, co-precipitation, and reconstruction). The organic drug and the Cu(II)-ibp were also immobilized by adsorption on LDH external surfaces. Materials were characterized by elemental analysis, UV/VIS, FTIR, and Raman spectroscopies, powder X-ray diffractometry (XRD), thermogravimetry, and electronic paramagnetic resonance (EPR). Mg3Al (ibp)(cop) (30% w/w of drug/material) and Mg3Al-(ibp)(ie) (33%) materials exhibit bilayer arrangements of ibp anions intercalated between the host layers. Mg3Al (ibp)(rec) and Mg3Al-(ibp)(ads) contain 13% and 6.2% of ibuprofenate, respectively. Mg3Al-(Cu-ibp)(ads) exhibits two Cu centers in different environments interacting with host layers. Pharmacological potential of materials are compared considering the amounts of immobilized drugs and their buffering properties. Mg3Al-(ibp)(ie) and Mg3Al-(ibp)(cop) exhibit poor buffering property, but contain high ibp amounts. Mg3Al-(ibp)(ads) despite having buffering property, contains low amount of ibuprofen. Mg3Al-(ibp)(rec) combines significant amount of immobilized ibp with good buffering property. Mg3Al-(Cu-ibp)(ads), due to the buffering property, may be an interesting new formulation aiming to decrease gastric irritation. PMID- 15793806 TI - Thermal and fractal analysis of diclofenac/Gelucire 50/13 microparticles obtained by ultrasound-assisted atomization. AB - The study describes the application of a spray-congealing technique, using a new ultrasound-assisted atomizer to prepare microparticles of diclofenac/Gelucire 50/13, with the aim to obtain a formulation of enhanced-release, at 10% w/w drug to-excipient ratio, without any employ of solvent. Scanning electron microscopy showed that it was possible to obtain almost spherically shaped and non aggregated microparticles; with good encapsulation efficiency (90% in most size fraction) and with a prevalent particle size in the range 150-350 mum. Image analysis results by SEM and the high fractal dimension value suggested that most particles have actually an ellipsoidal shape and a rather rough contour. Hot stage microscopy, differential scanning calorimetry, and X-ray powder diffractometry analysis were carried out to evaluate the nature of the solid state and the thermal behavior of the microparticles thus prepared. The in vitro tests displayed a significant increase of the diclofenac dissolution rate from ultrasound microparticles, compared with pure drug and with drug/Gelucire 50/13 physical mixtures. PMID- 15793808 TI - Pharmaceutical development of a lyophilised dosage form for the investigational anticancer agent Imexon using dimethyl sulfoxide as solubilising and stabilising agent. AB - Imexon is a member of the class of 2-cyanoaziridine derivatives, which have been of interest as immunomodulators and anticancer agents since the late 1970s. For the scheduled phase I clinical trials a stable, sterile, injectable pharmaceutical dosage form containing 100 mg Imexon was required. Despite adequate solubility, its instability in aqueous media seriously hampered the pharmaceutical development of Imexon. In this study we describe the successful use of the organic solvent dimethyl sulfoxide (DMSO) as a formulation vehicle for Imexon. DMSO is shown to provide the stability required for Imexon during manufacturing and to be a suitable vehicle for lyophilisation, which was employed to gain sufficient shelf-life for the final product. The relatively low vapour pressure of DMSO, which would theoretically result in extremely slow sublimation during lyophilisation, was shown not to limit the successful lyophilisation of Imexon from DMSO at a concentration of 25 mg/mL. The lyophilisation cycle developed for Imexon resulted in residual DMSO contents of 4.6 +/- 0.6% in the lyophilised product, limiting the amount of DMSO administered to the patient to well below the 50 mg/day acceptable in pharmaceutical products as stated in ICH guidelines. Imexon 100 mg/vial lyophilised product was shown stable for at least 12 months of storage at -20 degrees C and +5 +/- 3 degrees C in the dark. PMID- 15793809 TI - Multiple pathways are involved in the oral absorption of BMS-262084, a tryptase inhibitor, in rats: role of paracellular transport, binding to trypsin, and P glycoprotein efflux. AB - BMS-262084 is a potent and selective beta-lactam tryptase inhibitor with therapeutic potential for treating asthma. The oral bioavailability of BMS-262084 was low in rats (4% at a dose of 0.5 mg/kg) due to poor absorption. BMS-262084 was excreted mainly unchanged in the urine, suggesting minimal metabolism in rats. The objective of this study was to investigate the mechanisms of oral absorption of BMS-262084 in rats. Modulation of intestinal tight junctions, binding to trypsin, and involvement of the intestinal dipeptide transport system and P-glycoprotein (P-gp) in the absorption of BMS-262084 were examined. Coadministration of BMS-262084 with SQ-29852, a substrate of the intestinal dipeptide transport system, did not change the oral absorption of BMS-262084. An increase in the dose of BMS-262084 from 0.5 to 50 mg/kg resulted in a 3.7-fold increase in its oral absorption. Inulin absorption was enhanced upon coadministration with BMS-262084, suggesting the opening of tight junctions in the intestinal epithelium. Coadministration of aprotinin, a trypsin inhibitor, increased the oral absorption of BMS-262084 several fold. In vitro, using Caco-2 cells, BMS-262084 appeared to be a P-gp substrate, with an efflux ratio of 14. These results suggest that absorption of BMS-262084 is mediated by several concurrent mechanisms. At higher doses of BMS-262084, increased absorption may be primarily due to opening of tight junctions in the intestinal epithelium and consequent absorption via the paracellular pathway, while at lower doses, binding to trypsin may contribute to limiting its absorption. P-gp efflux may also play a role in influencing the absorption of BMS-262084. The intestinal dipeptide transporter system does not appear to be involved in the absorption of BMS 262084. PMID- 15793810 TI - Physicochemical and structural characterization of quercetin-beta-cyclodextrin complexes. AB - Quercetin is a bioactive flavonoid widely used as a health supplement. Being sparingly soluble and chemically unstable in aqueous intestinal fluids, quercetin is poorly absorbed orally. This study aimed to investigate the effects of three beta-cyclodextrins, namely, unsubstituted beta-cyclodextrin (beta-CD), hydroxypropyl-beta-cyclodextrin (HP-beta-CD), and sulfobutyl ether beta cyclodextrin (SBE-beta-CD) on the chemical stability and water solubility of quercetin, and to elucidate the complexation mechanisms of these beta-CDs with quercetin. Quercetin-beta-CD complexes in solution were characterized by stability assessment, phase solubility measurements, and 1H-nuclear magnetic resonance (NMR) spectroscopy. Molecular modeling was used to help establish the mode of interaction of the beta-CDs with quercetin. Solubility enhancements of quercetin obtained with the three beta-CDs followed the rank order: SBE-beta-CD > HP-beta-CD > beta-CD. The stability of quercetin at alkaline pHs also showed substantial improvement. NMR spectroscopic analysis suggested that the B-ring, C ring, and part of the A-ring of quercetin display favorable interaction with the hydrophobic cavity of the beta-CDs, which was confirmed by molecular dynamics (MD) simulations using a solvated model of the quercetin-beta-CD complex. An inclusion complex model has been established for explaining the observed augmentation of solubility and stability of quercetin in water by beta-CDs. PMID- 15793811 TI - Drug molecules as probes for studying the compatibility between gels and mucous tissue with dielectric spectroscopy. AB - In this interdisciplinary study low-frequency dielectric spectroscopy is used to assess the possibilities of intimate surface contact between a polymer gel and mucous tissue, which is generally considered to be the first step in the mucoadhesion process. The dielectric responses of six different gels, made from Carbopol 934 or Pluronic F-127 and containing 15 mM of a drug compound, either atenolol, alprenolol, or naproxen, were measured together with the responses of freshly excised porcine nasal mucosa and of systems made by combining the two. An analytical procedure is presented, which enables the dielectric response arising from the drug ions to be extracted. The drug ions are used as probes for measuring the ease, in terms of conductivity, with which they can pass the interface between the gel and the mucus layer. The results can be described by a compatibility factor that provides us with an assessment of the likelihood of intimate surface contact. The compatibility factors found in this study were generally higher for the Carbopol 934 gels than for the Pluronic F-127 gels, which is in agreement with the results of a previous study where sodium and chloride ions were used as probes for measuring the compatibility. Naproxen exhibited the largest difference in compatibility factor between the two gels; the highest compatibility factor of all systems was found for the Carbopol 934 gel whereas the gel based on Pluronic F-127 gave a value of approximately zero. This may partly be explained by interactions between the drug ions and the polymers. PMID- 15793812 TI - The effect of water to ethanol feed ratio on physical properties and aerosolization behavior of spray dried cromolyn sodium particles. AB - Cromolyn sodium (CS) was spray dried under constant operation conditions from different water to ethanol feed ratios (50:50-0:100). The spray dried CS samples were characterized for their physicochemical properties including crystallinity, particle size distribution, morphology, density, and water/ethanol content. To determine quantitatively the crystallinity of the powders, an X-ray diffraction (XRD) method was developed using samples with different crystallinity prepared by physical mixing of 100% amorphous and 100% crystalline CS materials. The aerodynamic behavior of the CS samples was determined using an Andersen Cascade Impactor (ACI) with a Spinhaler at an air flow of 60 L/min. Binary mixtures of each spray dried CS powder and Pharmatose 325, a commercial alpha-lactose monohydrate available for DPI formulations, were prepared and in vitro aerosol deposition of the drug from the mixtures was analyzed using ACI to evaluate the effect of carrier on deposition profiles of the spray dried samples. CS spray dried from absolute ethanol exhibited XRD pattern characteristic for crystalline materials and different from patterns of the other samples. The crystallinity of spray dried CS obtained in the presence of water varied from 0% to 28.37%, depending on the ratio of water to ethanol in the feed suspensions. All samples presented different particle size, water/ethanol content, and bulk density values. CS particles spray dried from absolute ethanol presented uniform elongated shape whereas the other samples consisted mainly of particles with irregular shape. Overall, fine particle fraction increased significantly (p < 0.01) with decreasing d50% and water and ethanol content of spray dried CS samples. Significant difference (p < 0.01) in deposition profiles of the drug were observed between corresponding carrier free and carrier blended formulations. The difference in deposition profiles of CS aerosolized from various spray dried samples were described according to the particle size, shape, and water/ethanol contents of the powders. The results of this study indicate that enhanced aerosol performance of CS can be obtained by spray drying of the drug from suspensions containing > or = 87.5% v/v ethanol. PMID- 15793813 TI - Inflammatory and immunological responses to subchronic exposure to endotoxin contaminated metalworking fluid aerosols in F344 rats. AB - Rats were exposed for 6 h per day in inhalation chambers to a 10 mg/m(3) concentration of metalworking fluid (MWF) contaminated with endotoxin at concentrations of 1813 (low dose) and 20,250 eu/m(3) (high dose) 5 days per week for 8 weeks. It was found that 94.7% of the MWF aerosol particles had diameters in the range of 0.42-4.6 microm, with geometric mean diameter of 1.56 microm. The body weight and pulmonary function parameters were measured every week during the 8 weeks of exposure, whereas bronchoalveolar lavage (BAL) fluid was prepared to measure the inflammatory markers and cytokines after the 8 weeks of exposure. There were no changes in body weight and respiratory function (tidal volume and respiratory frequency) during the 8 weeks of exposure to the MWF containing endotoxins, yet lung weight increased significantly (P < 0.05) in the rats exposed to the MWF both with and without endotoxins. The number of polymorphonuclear (PMN) cells in the BAL fluid increased significantly (P < 0.05) in the rats exposed to MWF with endotoxins, and the levels of cytokines such as IL-4, INF-gamma, IL-1beta, and TNF-alpha also were significantly increased (P < 0.05) compared to the control. The NOx production activity of the BAL cells increased significantly (P < 0.05) in the rats exposed to the MWF with and without endotoxins. Increases in lung weight, number of PMN cells, and levels of extracellular cytokines and NOx were all more significant in the rats exposed to the MWF with endotoxins rather than in those exposed to MWF without endotoxins. In spleen cell cultures, T-cell proliferation activity was decreased, yet cytokine levels (INF-gamma, IL-1beta, IL-4, and TNF-alpha) remained unchanged after repeated exposure to MWF with and without endotoxins. Although the levels of total IgG(1), IgG(2a), and IgE antibodies in the serum were not changed, the levels of endotoxin-specific antibodies, including IgG(2a) and IgE, were increased significantly (P < 0.05) in the rats exposed to endotoxins, but there was not a significant increase in endotoxin-specific IgG(1). When taken together, the results indicate that lung inflammatory responses can be induced without changing pulmonary function after repeated exposure to MWFs contaminated with endotoxins. In addition, endotoxin-specific IgG(2a) and IgE may be effective biomarkers for workers exposed to MWFs contaminated with endotoxins in the workplace. PMID- 15793814 TI - Predictive risk thresholds for survival protection of farmed abalone, Haliotis diversicolor supertexta, exposed to waterborne zinc. AB - Using a probabilistic risk-based framework, we have developed a simple predictive risk threshold model for protecting the survival of farmed abalone, Haliotis diversicolor supertexta, exposed to waterborne zinc (Zn). Probabilistic techniques using Monte Carlo analysis propagate parameter uncertainty/variability throughout the model, providing decision makers with a credible range of information and increased flexibility in establishing a specific Zn level in aquacultural ecosystems. We coupled a first-order two-compartment bioaccumulation model with a reconstructed dose-response profile based on a three-parameter Hill equation model to form a probabilistic risk model in order to determine the risk quotient associated with a 10% probability of exceeding the abalone 5% effect concentration (EC(5)) at site-specific abalone farms. Sensitivity analysis revealed that waterborne Zn concentration (C(w)) and algae bioconcentration factor (BCF(a)) have a significant effect on Zn levels in abalone. Using multiple nonlinear regression analysis with C(w) and BCF(a) as the parameters, a predictive risk threshold equation that can be used in a variety of site-specific conditions was developed for protecting the survival of farmed abalone. We believe this probabilistic framework provides an effective method for conceptualizing a public policy decision vis-a-vis the establishment of a specific acceptable risk threshold for aquacultural water quality management. PMID- 15793816 TI - Effects of interactions between cadmium and zinc on phytochelatin and glutathione production in wheat (Triticum aestivum L.). AB - It has been proposed that phytochelatins (PCs) act as a biomarker for the evaluation of metal toxicity. Little attention has been paid to the effects on metal combinations and glutathione (GSH), the most abundant cellular thiol. In the present study the effects of interactions between cadmium (Cd) and zinc (Zn) on PC and GSH production were examined in wheat tissue over 14 days' exposure. The results showed that the presence of Zn alleviated Cd toxicity, accompanied by a reduction of Cd uptake. Cd and Zn exposure increased PC-SH levels in concentration-, tissue- and time-dependent manners. Of the two metals, Cd was more effective than Zn in PC-SH production. Interactions of Cd and Zn with respect to PC-SH production may be synergistic or inhibitory, strongly depending on duration of exposure and concentration of the metal combinations. Cd also stimulated GSH production in concentration-, tissue- and time-dependent manners, whereas Zn had no significant effects on GSH levels. Compared to the presence of Cd alone, the presence of Zn reduced GSH levels in a tissue-dependent manner over the growth period. The results of the study suggest that metal interactions should be highly considered in the application of PCs and GSH as potential biomarkers for the evaluation of metal toxicity, as most metal-polluted natural environments are contaminated with more than one metal. PMID- 15793815 TI - In vitro percutaneous penetration of acyclovir from solvent systems and Carbopol 971-P hydrogels: influence of propylene glycol. AB - The mechanism underlying propylene glycol (PG) effects on acyclovir (ACV) penetration through human epidermis were studied. Solvent systems and Carbopol gels containing increasing percentage of PG (from 0% to 70%, w/w) were used. Viscosity studies of both vehicles were carried out to characterise the influence of rheological behaviour. In solvent systems skin permeation values of ACV increase as the concentration of PG increase yielding a maximum enhancement ratio (ER = 10) for 70% PG. The release rate of ACV from gels was determined. Higuchi's model was used to estimate the apparent diffusion coefficient of the drug. These values show a decrease as the content of PG in the vehicle increases; this effect could be attributed to the increase of the viscosity in the diffusional pathway. When gels are used skin permeation values of ACV were smaller than those of the solvent systems. This could be attributed to the network structure created by the polymer that increases the length of the diffusional pathway. The maximum ER (= 6.8) was for Carbopol gel containing 50% PG. Therefore, these gels can be considered candidates for further research to confirm their usefulness as delivery systems for ACV topical formulations. PMID- 15793819 TI - Copper toxicity and bioaccumulation in Chinese cabbage (Brassica pekinensis Rupr.). AB - Copper is among the major heavy metal contaminants in the environment with various anthropogenic and natural sources. Human health risk from heavy metal bioaccumulation in vegetables has been a subject of growing concern in recent years. To investigate Cu phytotoxic effects and bioaccumulation in the popular vegetable Chinese cabbage (Brassica pekinensis Rupr) as well as the implications for human health due to Cu in the vegetable supply, seed germination and pot culture experiments with this vegetable were carried out. Six levels (0, 0.008, 0.031, 0.125, 0.5, and 2.0 mM/L) and 3 levels (0, 0.2, and 1.0 mM/kg) of Cu treatments were performed for the seed germination and pot culture experiments, respectively. The LC(50) of Cu for seed germination of Chinese cabbage was 0.348 mM/L. In the pot culture experiments, Cu treatments significantly increased electrolyte leakage and peroxidase activity of shoot tissues, demonstrating Cu phytotoxicity to the plants. On the other hand, Cu treatments significantly stimulated, instead of reduced, chlorophyll content. Cu treatments did not show a significant effect on shoot biomass. Compared to the control, Cu treatments significantly elevated the Cu content of the shoots-9.9, 42.5, and 119.0 mg/kg (DW) of Cu were detected in the 0, 0.2, and 1.0 mM/kg treatments, respectively. These results showed that although the plants accumulated an elevated copper content and suffered damage to some extent under Cu treatment, they looked healthy. It was suggested that Chinese cabbage with an elevated Cu content and without showing visible symptoms of damage possibly could cause a risk to human health from the transfer of the metal in food. PMID- 15793820 TI - Immune responses in mice of beta-galactosidase adsorbed or encapsulated in poly(lactic acid) and poly(lactic-co-glycolic acid) microspheres. AB - The immune response induced in mice by beta-galactosidase (beta-gal) adsorbed or encapsulated on poly(lactic acid) (PLA) and poly(lactic-co-glycolic acid) (PLGA) microspheres was investigated. The encapsulated protein elicited higher antibody response than the protein adsorbed on the microspheres in the case of the PLA microspheres. However, the encapsulated protein elicited weaker antibody response than the adsorbed protein in the case of the PLGA (50:50) microspheres, probably because, in this case, the encapsulation process adversely affected protein immunogenicity. In the case of adsorbed beta-gal, higher antibody response was obtained with the PLA microspheres than with the PLGA (50:50) microspheres. This may be related to the lower rate of beta-gal desorption from the PLA microspheres. Based on the immunoglobulin G1/immunoglobulin G2a ratios and the stimulation indices for interferon-gamma and interleukin-4, beta-gal encapsulated or adsorbed on PLA microspheres induced a Th(1)-biased immune response whereas beta-gal encapsulated or adsorbed on PLGA (50:50) microspheres induced a Th(2) biased immune response. The results obtained indicate that more potent immune responses are obtained when the protein is encapsulated than adsorbed on the microspheres, providing that the encapsulation process does not adversely affect protein immunogenicity. Also, the type of polymer used to prepare the microspheres, but not the method of protein association with the microspheres, may affect the type of immune response. PMID- 15793821 TI - Study on the self-setting property and the in vitro bioactivity of beta-Ca2SiO4. AB - This study sought to investigate the physical and chemical properties of beta dicalcium silicate (beta-Ca(2)SiO(4)) in order to evaluate its use as an injectable bioactive cement filler. Workable beta-Ca(2)SiO(4) pastes with a liquid-to-powder (L/P) ratio of 1.0-1.2 could be injected for 10-30 min (nozzle diameter 2.0 mm) and enabled initial setting times of 60-180 min. The setting process yielded cellular structures with compressive strengths of 4.8-28.8 MPa after 2-28 days. The paste was soaked in simulated body fluid (SBF), and the results demonstrated that it exhibited a moderate degradation and could induce carbonated hydroxyapatite formation. The ionic products of the paste dissolution enhanced a proliferative response of fibroblasts compared with the cells cultured alone, and this cement could also support adhesion and spreading of the mesenchymal stem cells. Finally, with the use of gentamicin as a model drug, it was found that a high dose of drug release from the paste was maintained for 14 days, and there was a sustained release over 4 weeks. This combination of properties indicates that the novel beta-Ca(2)SiO(4) cement might be suitable for potential applications in the biomedical field, preferentially as materials for bone/dental repair and controlled drug-delivery systems. PMID- 15793822 TI - Selective control of toxic Microcystis water blooms using lysine and malonic acid: an enclosure experiment. AB - Three enclosures (10 x 10 x 1.5-1.3 m in depth) were set beside Dianch Lake, Kunming, People's Republic of China, for the period from July 28 to August 26, 2002. The enclosures were filled with cyanobacterial (Microcystis aeruginosa) water bloom-containing lake water. Lake sediment that contained macrophytes and water chestnut seeds was spread over the entire bottom of each enclosure. Initially, 10 g/m(2) of lysine was sprayed in Enclosure B, and 10 g/m(2) each of lysine and malonic acid were sprayed together in Enclosure C. Enclosure A remained untreated and was used as a control. The concentrations of lysine, malonic acid, chlorophyll a, and microcystin as well as the cell numbers of phytoplankton such as cyanobacteria, diatom, and euglena were monitored. On day 1 of the treatment, formation of cyanobacterial blooms almost ceased in Enclosures B and C, although Microcystis cells in the control still formed blooms. On day 7 Microcystis cells in Enclosure B that had been treated with lysine started growing again, whereas growth was not observed in Microcystis cells in Enclosure C, which had been treated with lysine and malonic acid. On day 28 the surface of Enclosure B was covered with water chestnut (Trapa spp.) and the Microcystis blooms again increased. In contrast, growth of macrophytes (Myriophllum spicatum and Potamogeton crispus) was observed in Enclosure C; however, no cyanobacterial blooms were observed. Lysine and malonic acid had completely decomposed. The microcystin concentration on day 28 decreased to 25% of the initial value, and the pH shifted from the initial value of 9.2 to 7.8. We concluded that combined treatment with lysine and malonic acid selectively controlled toxic Microcystis water blooms and induced the growth of macrophytes. PMID- 15793823 TI - Toxic effects of Al-based coagulants on Brassica chinensis and Raphanus sativus growing in acid and neutral conditions. AB - The ecotoxicological effects of aluminum (Al)-based coagulants are of concern because of their wide-ranging applications in wastewater treatment and water purification. As important Al-based coagulants, AlCl(3) and PAC (polyaluminum chloride) were selected as examples to examine the toxic effects on representative vegetables including the cabbage Brassica chinensis and the radish Raphanus sativus over a range of exposure concentrations in neutral (pH 7.00) and acidic (pH 4.00) conditions, using seed germination and root elongation in the early-growth stage as indicators of toxicity. The results showed that root elongation of the two vegetables was a more sensitive indicator than was seed germination for evaluating the toxicity of Al. As a single influencing factor, H(+) had no significant direct effects on root elongation of Brassica chinensis and Raphanus sativus under the experimental conditions. The toxicity of Al played the main role in inhibiting root elongation and seed germination and was strongly related to changes in pH. There was a markedly positive relationship between the inhibitory rate of root elongation, seed germination, and the concentration of Al at pH 4.00 (p < 0.01). The toxic effect of AlCl(3) on Brassica chinensis was less with a neutral pH than at pH 4.00, but Raphanus sativus was more susceptible to AlCl(3) toxicity at a neutral pH than at pH 4.00. Both Raphanus sativus and Brassica chinensis had a more toxic response to a low concentration (<64 mg . L( 1)) of PAC in a neutral condition than in an acidic condition. Undoubtedly, the Al toxicity caused by Al-based coagulants at a neutral pH is relevant when treatment solids are used in agriculture. PMID- 15793824 TI - Toxicities of triclosan, phenol, and copper sulfate in activated sludge. AB - The effect of toxicants on the BOD degradation rate constant was used to quantitatively establish the toxicity of triclosan, phenol, and copper (II) against activated sludge microorganisms. Toxicities were tested over the following ranges of concentrations: 0-450 mg/L for phenol, 0-2 mg/L for triclosan, and 0-35 mg/L for copper sulfate (pentahydrate). According to the EC(50) values, triclosan was the most toxic compound tested (EC(50) = 1.82 +/- 0.1 mg/L), copper (II) had intermediate toxicity (EC(50) = 18.3 +/- 0.37 mg/L), and phenol was the least toxic (EC(50) = 270 +/- 0.26 mg/L). The presence of 0.2% DMSO had no toxic effect on the activated sludge. The toxicity evaluation method used was simple, reproducible, and directly relevant to activated sludge wastewater treatment processes. PMID- 15793825 TI - Expression of carotenoid pigments of haloarchaeal cultures exposed to aniline. AB - The effects of exposure to aniline on growth and pigmentation in three haloarchaeal isolates from the Indian subcontinent--GUSF (MTCC 3265), from the estuarine saltpans of Goa, India; and GURFT-1 and GURFP-1, both from continental shelf sediments of the west coast of India--were studied. In nutrient-rich tryptone yeast extract medium containing 25% NaCl/crude salt, the growth of GUSF, measured as absorbance at 600 nm, was not affected significantly at all concentrations of aniline used [0.005%-0.04% (v/v)], whereas the growth phases of GURFT-1 and GURFP-1 were affected at concentrations > 0.005%; the total yield, however, was nearly equal to the yield of cultures growing in the absence of aniline. GURFT-1 reached approximately 40% of total yield on the 7th day in the presence of 0.04% aniline, which declined thereafter. The pigmentation observed visually was completely abolished at concentrations of aniline greater than 0.01%. Spectral scans of acetone extracts of the pigment of each of the cultures exposed to concentrations of aniline > 0.01% showed that (i) the bacterioruberin component of the pigments (absorbance in the range 390-500 nm) was completely abolished and (ii) the pigment component(s) shifted toward squalene and phytofluene derivatives (320-360 nm). This is the first report examining the effect of an aromatic pollutant such as aniline on the growth and pigmentation of haloarchaeal cultures. PMID- 15793826 TI - Histopathological alterations in the edible snail, Babylonia areolata (spotted babylon), in acute and subchronic cadmium poisoning. AB - Histopathological alterations in 6- to 8-month-old juvenile spotted babylon, Babylonia areolata, from acute and subchronic cadmium exposure were studied by light microscopy. The 96-h LC(50) value of cadmium for B. areolata was found to be 3.35 mg/L, and the maximum acceptable toxicant concentration (MATC) was 1.6 mg/L. Snails were exposed to 3.35 and 0.08 mg/L (5% of MATC) of cadmium for 96 h and 90 days, respectively. After exposure the gill, the organs of the digestive system (proboscis, esophagus, stomach, digestive gland, and rectum), and the foot were analyzed for cadmium accumulation. The results showed that most digestive organs had a high affinity for cadmium. The main target organ was the stomach, which could accumulate on average 1192.18 microg/g dry weight of cadmium. Cadmium was shown to accumulate to a lesser extent in the digestive gland, gill, rectum, esophagus, proboscis, and foot. Histopathological alterations were observed in the gill and digestive organs (proboscis, esophagus, stomach, and rectum). The study showed that the stomach and gill were the primary target organs of both acute and subchronic exposure. Gill alterations included increased size of mucous vacuoles, reduced length of cilia, dilation and pyknosis of nuclei, thickening of basal lamina, and accumulation of hemocytes. The epithelial lining of the digestive tract showed similar alterations such as increased size of mucous vacuoles, reduced length of cilia, and dilation of nuclei. In addition, fragmentation of the muscle sheath was observed. PMID- 15793827 TI - Acute toxicity of copper to the threespine stickleback, Gasterosteus aculeatus. AB - This study focuses on characterizing the acute toxicity of copper in freshwater to the threespine stickleback, Gasterosteus aculeatus, a small and widely distributed euryhaline fish. The threespine stickleback is used as an effluent monitoring species in both Canada and the United States, yet in some locations natural populations are listed as threatened or endangered. Four 96-h static renewal acute toxicity tests were performed in moderately hard water using U.S. EPA methods with adult fish (mean wet weight = 0.41 g/fish). The geometric mean of the 24-, 48-, 72- and 96-h LC(50)s based on measured concentrations of total copper (estimated dissolved copper in parentheses) in the test solutions were 382.2 (366.9), 278.7 (267.6), 256.6 (246.3), and 227.2 (218.1) microg Cu/L, respectively. Conservative estimates of acute toxicity thresholds, made using LC1 values, for adult threespine sticklebacks over 24-, 48-, 72- or 96-h exposure periods in moderately hard water are approximately 114.3 (109.7), 78.3 (75.2), 67.0 (64.3), and 52.4 (50.3) microg Cu/L, respectively. Test results were normalized to a range of water hardness from very soft to very hard using two U.S. EPA methods, the water hardness and the Biotic Ligand Model normalization procedures. Subsequently, interspecies sensitivity comparisons were made with aquatic animal species used in both the current and proposed U.S. EPA copper water quality criteria documents. Information reported in this study may be useful in effluent toxicity identification evaluations, ecological risk assessments and criteria development where copper is a concern. PMID- 15793828 TI - Influence of drinking water composition on quantitation and biological activity of dissolved microcystin (cyanotoxin). AB - Toxic cyanobacteria in aquatic environments have been implicated in many poisoning incidents of livestock, wildlife, and domestic animals. Microcystins (MCYSTs) in water supplies represent a risk to public health. This work investigated the effect of water composition on the quantitation and biological activity of MCYSTs analyzed by different methods (HPLC, ELISA, and protein phosphatase 1 inhibition assay). Different MCYST concentrations were added to deionized water and quantified, confirming the efficiency of these analytical methods. MCYST concentrations diluted in drinking water had reduced detection by all methods tested. The drinking water used contained a free chlorine concentration of 2.5 mg/L and an Fe concentration of 0.45 mg/L, and the conductivity was 69.8 microS cm(-1), whereas in deionized water, free chlorine and Fe were not detectable, and the conductivity was 1.6 microS cm(-1). Drinking water also interfered with the biological activity of MYCSTs, as these toxins showed reduced protein phosphatase-1 inhibition. A free chlorine concentration of 2.5 mg/L in deionized water was completely effective in preventing any detection of 10 microg/L of added MCYSTs. Fe and Al ions also were very effective in reducing MCYST detection. The chemical composition of drinking water thus affected MCYST detection, indicating a significant reduction in quantitation of this molecule either because of its decomposition or through complexation with metal ions. PMID- 15793829 TI - Life-cycle exposure of fathead minnows (Pimephales promelas) to an ethinylestradiol concentration below 1 ng/L reduces egg fertilization success and demasculinizes males. AB - Forty-eight hours after fertilization, fathead minnow (Pimephales promelas) eggs were exposed to the synthetic estrogen 17alpha-ethinylestradiol (EE2) at nominal concentrations of 0.32 and 0.96 ng/L and measured concentrations of 3.5, 9.6, and 23 ng/L. The fish were observed through the larval, juvenile, and adult stages. Growth, secondary sex characteristics, the liver somatic index, the gonadosomatic index, and fecundity were examined after several lengths of exposure. No significant changes were seen in fry or juvenile growth from 8 to 30 days posthatch (dph). An increase in the ovipositor index (a female secondary sex characteristic) was the most sensitive early response at 60 dph and was seen in fish exposed to EE2 concentrations > or = 3.5 ng/L. Continuation of the EE2 exposure until 150 dph, through maturation and reproduction, allowed measurement of two sensitive end points: decreased egg fertilization and sex ratio (skewed toward females), both of which were significantly affected at the lowest EE2 concentration tested, 0.32 ng/L. The next most sensitive end point was demasculinization (decreased male secondary sex characteristic index) of males exposed to an EE2 concentration of 0.96 ng/L. The effects of low concentrations of EE2 (0.32 and 0.96 ng/L) were manifested in male fish (decreased male sex characteristics and reduced egg fertilization success), whereas female fish showed no changes in the gonadosomatic index. Exposure to higher EE2 concentrations negatively affected females, as shown by a reduced gonadosomatic index at 150 dph in fish exposed to > or =3.5 ng/L EE2. Although there were some end points that showed changes at 60 dph, the reproductive end points and external sex characteristics measured in mature fish at 150 dph were more sensitive, with response thresholds of EE2 ranging from 0.32 to 0.96 ng/L. The concentrations of EE2 that negatively affected fathead minnows were similar to or lower than those detected in many municipal wastewater effluents. In conclusion, life-cycle exposure of fathead minnows proved to be a very sensitive bioassay, and responses were seen at concentrations of less than 1 ng/L, which are environmentally relevant concentrations of EE2. PMID- 15793830 TI - Prevention and treatment of amputation neuroma by an atelocollagen tube in rat sciatic nerves. AB - To evaluate the potential of the atelocollagen tube as a cap for amputation neuromas, the histological and histochemical characteristics of the neuroma and spinal cord were compared with those following silicone capping. Four weeks after the transection of 18 rat sciatic nerves, the amputated neuroma was resected, and the nerve stump inserted into an atelocollagen or silicone tube. The histological changes in the nerve ends and c-fos expression in the dorsal horn of the fourth lumbar spinal cord were evaluated at 4 weeks postoperatively. The regenerated nerve structure in the atelocollagen or silicone tube was very thin. In contrast, a typical bulbous neuroma was observed in the control group (the nerve stump was left in place). The atelocollagen and silicone tube groups demonstrated fewer c fos-expressed cells in the spinal cord than the controls. These results suggest that capping by an atelocollagen tube, like that by a silicone tube, might successfully prevent an amputated neuroma from forming, and suppress induced pain. The atelocollagen tube may be a promising biomaterial for the prevention or treatment of a painful amputation neuroma. PMID- 15793831 TI - A survey of potential stressor-induced physiological changes in carp (Cyprinus carpio) and barbel (Barbus bocagei) along the Tajo River. AB - The objective of this study was to evaluate fish response to acute stress induced by confinement after capture. Because of the previously reported presence of chemical compounds in the Tajo River basin where the study samplinig took place, an exposure biomarker to organic chemicals (retinol) was used. Cortisol and glucose were used as stress biomarkers. Plasma levels of cholesterol were used as lipidic metabolism indicators, and retinol level was used as a specific exposure biomarker. A reference site was established along 300 km of the Tajo River, and nine sampling sites were selected on the basis of whether various human activities and hydrographic characteristics were present. A total of 55 carp (Cyprinus carpio) and 52 barbel (Barbus bocagei) were examined. Cortisol and glucose levels were considered acceptable indicators of the response of the fish to induced stress. In the barbel, plasma retinol levels decreased at two of the sampling sites indicating possible exposure to organic compounds. The overall evaluation of these parameters enabled us to identify three sampling sites at which more studies should be carried out. The possible relationship between the health state of wild fish and the presence of organic compounds or sources of pollution was considered. PMID- 15793832 TI - Acrofacial dysostosis type Rodriguez. AB - The acrofacial dysostoses (AFD) are a clinically and causally heterogeneous group of conditions characterized by mandibulofacial dysostosis and a variety of limb anomalies. Several abnormalities affecting different internal organs and the central nervous system (CNS) have been described. Depending on the type of limb defects, two major groups have been delineated: (1) with predominantly pre-axial anomalies, Nager type AFD, and (2) with predominantly post-axial involvement, Genee-Wiedemann form of AFD, also known as POADS, respectively. Other forms of "true AFD" have been described as Kelly, Reynolds, Arens (also Tel Aviv form), Rodriguez (or Madrid form), Richieri-Costa, and Patterson-Stevenson-Fontaine types. However, whether they are distinct entities or represent variants of the same condition remains unclear. Rodriguez AFD was described as a new lethal form of AFD in three affected sibs with severe mandibular hypoplasia, severe predominantly pre-axial limb deficiencies, absent fibulae and ribs, and internal organ anomalies, the most remarkable of which are arrhinencephaly and abnormal lung lobulation. We present a newborn girl with Rodriguez type of AFD, who died a few days after the birth due to respiratory failure. The phenotype and the cause of this condition are discussed. PMID- 15793833 TI - Ultrastructural analysis of guided nerve regeneration using progesterone- and pregnenolone-loaded chitosan prostheses. AB - Recently, numerous guide chambers for the treatment of injured nerves made up of different biomaterials have been designed, capable of hosting living cells or carrying neurotrophic or neuroactive substances to be directly released to the injured tissue. In this study, chitosan prostheses containing neurosteroids (progesterone and pregnenolone) were used for bridging a 10-mm gap in the rabbit facial nerve. Gas chromatography was used to quantify neurosteroid content in the prostheses prior to and after subcutaneous implantation at different periods of up to 60 days. The regeneration of the nerve fibers were evaluated at 15 and 45 days after axotomy by means of ultrastructural morphometric analysis. Different nerve fibers regenerative patterns were seen depending the groups studied and the analyzed stages. At 15 days after axotomy, the newly regenerating tissue revealed Schwann cells holding nonmyelinated nerve fiber bundles in an incipient and organized regenerative pattern. At 45 days, the regenerating tissue showed myelinated nerve fibers of different sizes, shapes, and myelin sheath thickness. Although the regeneration of the nerve fibers under neurosteroid treatment showed statistically significant differences in comparison with vehicle regenerated tissue, progesterone-loaded chitosan prostheses produced the best guided nerve regeneration response. These findings indicate that chitosan prostheses allowed regeneration of nerve fibers in their lumen, and when containing neurosteroids produced a faster guided nerve regeneration acting as a long-lasting release delivery vehicle. PMID- 15793834 TI - Malpuech syndrome: three patients and a review. AB - We describe three patients with Malpuech syndrome from two families. Previously, 10 patients from 6 families have been reported. Consanguinity in two families suggests autosomal recessive inheritance. Growth retardation, mental retardation, cleft lip, and/or palate, hypertelorism, urogenital abnormalities, and caudal appendage are the key features. Although the spectrum of the features in the reported patients is variable, we do think this syndrome represents a distinct entity. Chromosomal anomalies should be carefully searched for. We discuss differential diagnosis and possible candidate genes and propose diagnostic criteria for Malpuech syndrome. PMID- 15793836 TI - A girl with inverted triplication of chromosome 3q25.3 --> q29 and multiple congenital anomalies consistent with 3q duplication syndrome. AB - We report a newborn girl with intrachromosomal triplication of 3q25.3 --> q29 (mosaicism) who died at the age of 3.5 weeks due to her malformations. She demonstrated disproportionate short stature with short limbs, a prominent and hairy forehead, thick eyebrows, synophrys, small upturned nose, full cheeks, micrognathia, and low set malformed and posteriorly rotated ears, short and webbed neck, hydrocephalus, Dandy-Walker malformation, spina bifida, complex heart defect (ventricular and atrial septal defect, malrotation, and interrupted aortic arch), omphalocele, polycystic kidneys, postaxial polydactyly of left hand, and generalized hirsutism; all signs have been associated with the dup(3q) syndrome previously. The facial appearance (hairy forehead, thick eyebrows, synophrys, small upturned nose, full cheeks, micrognathia, low set malformed and posteriorly rotated ears) showed resemblance to the Brachmann-de Lange syndrome (BDLS), but the patient did not fulfill the diagnostic criteria for BDLS. There has been only one report of a direct triplication of chromosome 3 until now, but in our case the triplicated area is larger, located more proximally, and includes the hypothetical BDLS critical gene region-CDL1. Our findings lend support to distal chromosome 3q, or chromosome 3q26.3, comprises the critical area for the dup(3q) phenotype resembling the BDLS. PMID- 15793835 TI - Somatic and gonadal mosaicism in Hutchinson-Gilford progeria. AB - We have studied a patient with Hutchinson-Gilford progeria (HGP). Sequence analysis of the LMNA gene demonstrated the presence of a c.1824 C > T (p.G608G) mutation, activating a cryptic splice donor site and leading to the formation of a truncated Lamin A protein. All molecularly characterized autosomal dominant HGP cases described so far result from de novo LMNA mutations, mostly originating on the paternal allele and are often linked with advanced paternal age. However, in our patient, the mutation was transmitted by the mother who showed somatic and germline mosaicism without HGP manifestations. PMID- 15793837 TI - Fryns syndrome with osteochondrodysplasia. PMID- 15793838 TI - Barth syndrome: TAZ gene mutations, mRNAs, and evolution. AB - Barth syndrome (MIM 302060) is an X-linked condition that includes dilated cardiomyopathy, neutropenia, failure to thrive, abnormal mitochondria, and 3 methylglutaconic aciduria. The mutated gene, TAZ, first described in 1996, appeared to produce a large set of alternatively spliced mRNAs with initiations of transcription upstream of exons 1 and 3. Since then, disease-causing mutations have been found in all exons including, most recently, a missense mutation in the controversial exon 5. Because of the initially described second initiation of transcription in intron 2, with in-frame initiation of translation in exon 3, we hypothesized that subjects with mutations in exons 1 and 2 would produce more normal "short product" that might attenuate their phenotype. Moreover, it was of interest to determine which splice variants were potentially functional as exon 5 is not present in yeast and rodents, and the variant lacking this exon is the most abundant. Using RT-PCR, we characterized TAZ mRNAs in cultured lymphocytes from nine subjects with Barth syndrome and two healthy controls. The TAZ genes and mRNAs of primates were also included. We found the following: (1) there is only one site for initiation of transcription, and the normal alternatively spliced assortment is limited to full-length, delta5, delta7, delta5delta7; (2) there are two alternative splice sites within introns 1 and 2 that could potentially produce an in-frame product; (3) exon 5 evolved into "exonhood" in the primate lineage after the split between Old World monkeys and hominoid primates; and (4) our results suggest that only two functional protein variants exist in lymphocytes: delta5 and full-length. Although exon 5 does not appear to be required for TAZ function in yeast and monkeys, its evolution to a highly conserved spliced exon in hominoid primates and the recent finding of an exon 5 mutation in a patient with Barth syndrome suggest that the full-length variant is important to TAZ function. PMID- 15793839 TI - Tenascin-X deficiency in autosomal recessive Ehlers-Danlos syndrome. AB - We present two unrelated individuals with complete deficiency of tenascin-X, resulting in an autosomal recessive form of Ehlers-Danlos syndrome (EDS). Consistent with the original description of tenascin-X deficiency, these individuals had marked skin hyperextensibility, easy bruising, and joint laxity. Unlike classical EDS they did not have atrophic scarring or poor wound healing. Significant medical problems occurring in these individuals included severe diverticular intestinal disease, mitral valve prolapse requiring valve replacement, and obstructive airway disease. PMID- 15793840 TI - Peginterferon alfa-2b plus ribavirin treatment in children and adolescents with chronic hepatitis C. AB - Peginterferon plus ribavirin is standard therapy for adults with chronic hepatitis C. As no data are available for children, the aim of the study was to evaluate the efficacy and tolerability of peginterferon alfa-2b in combination with ribavirin in chronically infected children. Genotypes, alanine aminotransferase levels, and different routes of viral transmission were considered. In an open-labeled, uncontrolled pilot study, 62 children and adolescents (range, 2-17 years) were treated with subcutaneous peginterferon alfa 2b at a dose of 1.5 microg/kg body weight once per week plus oral ribavirin (15 mg/kg x day) for 48 weeks. Sixty-one patients completed the study. Twenty-three children discontinued therapy after 6 months according to study protocol. Sustained viral response was documented in 22 (47.8%)of 46 patients with genotype 1, in 13 (100%) of 13 with genotype 2 or 3, in 1 of 2 with genotype 4, in 19 (70.4%) of 27 children with parenteral, in 12 (48%) of 25 with vertical, and in 5 of 9 with unknown route of infection. Overall, treatment was well tolerated. Nevertheless, some side effects were present in all treated patients. Eighty three percent had leucopenia, but only 3 individuals required dose reduction and 10.3% developed thyroid autoantibodies and thyroid dysfunction. In conclusion, combination treatment of peginterferon alfa-2b with ribavirin showed encouraging results and was well tolerated in children and adolescents with chronic hepatitis C. Weekly dosing of peginterferon alfa-2b is a considerable advance for this age group. The treatment is not approved for children. Further controlled trials are needed. PMID- 15793841 TI - Functional disomy of Xp: prenatal findings and postnatal outcome. AB - We report on trisomy of the short arm of the X chromosome (Xp11.2 --> pter) due to a de novo unbalanced X;13 translocation diagnosed prenatally in a female fetus. Amniocentesis was performed at 20-weeks' gestation following ultrasound finding of a Dandy-Walker malformation. The trisomy of Xp11.2 --> pter was confirmed with fluorescence in situ hybridization (FISH), using an X chromosome painting probe and telomeric FISH probes specific for the short arm of chromosome X. The karyotype was defined as 46,XX,der(13)t(X;13)(p11.2;p11.2). Molecular analysis suggested that the extra Xp material was of paternal origin. FISH analysis with an XIST probe showed that the derivative chromosome 13 did not include the XIST locus at the X-inactivation center (XIC). A complex phenotype was seen at birth including macrosomia, facial dysmorphism with preauricular tag, congenital heart defects, and structural brain malformations. Because the derivative chromosome was not subject to X inactivation, functional disomy of Xp11.2 --> pter most likely accounts for the abnormal phenotype in this patient. PMID- 15793842 TI - Unusual intestinal lamellae in the nematode Rhabditophanes sp. KR3021 (Nematoda: Alloinematidae). AB - The free-living nematode Rhabditophanes sp. has recently been placed in a clade of animal parasites and may be a unique example of a reversal to a nonparasitic lifestyle. Detailed morphological analysis of the intestine reveals the unusual and unique structure of splitting microlamellae forming a meshwork with cavities along the entire intestinal tract. Secretion vesicles were observed along the whole tract and along the length of the lamellae. It is suggested that these lamellae are adaptations to a different digestive strategy where low food availability and a low absorption surface are compensated for by maximizing the nutrient uptake efficiency along the entire length of the intestine. The likely reversal to a free-living life cycle may have caused drastic changes in diet, providing the necessary driving forces to such morphological changes. PMID- 15793843 TI - Iron- and inflammation-induced hepcidin gene expression in mice is not mediated by Kupffer cells in vivo. AB - Hepcidin, a recently discovered iron regulatory peptide, is believed to inhibit the release of iron from absorptive enterocytes and macrophages. Liver hepcidin synthesis is induced in vivo by iron stores and inflammation. The molecular basis of the regulation of hepcidin gene expression by these effectors in hepatocytes is currently unknown, although there is strong evidence that indirect mechanisms are involved. The aims of this study were to gain insight into these mechanisms and to determine to what extent other liver cell types are responsible for transducing the signal by which hepcidin expression is regulated in mouse hepatocytes. For this, we depleted Kupffer cells by injection of liposome encapsulated clodronate and then studied iron- and inflammation-induced hepcidin gene expression. In addition, we directly evaluated the role of the inflammatory cytokine interleukin 6 (IL-6) by using IL-6-deficient mice. Our results show that iron is able to induce hepcidin gene expression independently of Kupffer cells in the liver and circulating IL-6. In contrast, we show that hepcidin gene induction by inflammation is also independent of Kupffer cells, but involves, at least partly, IL-6. In conclusion, these results show that two independent regulatory pathways control hepcidin gene expression and suggest that hepatocytes play a key role in the regulation of hepcidin gene expression by sensing iron and inflammatory signals. PMID- 15793844 TI - A novel transthyretin mutation V32A in a Chinese man with late-onset amyloid polyneuropathy. AB - We report a Chinese patient with amyloidotic polyneuropathy associated with a novel transthyretin mutation (V32A). He presented with slowly progressive sensorimotor polyneuropathy accompanied by autonomic dysfunction and cardiomyopathy by echocardiography. This mutation is likely to be associated with late onset and low-penetrance phenotype. PMID- 15793845 TI - Lambert-Eaton myasthenic syndrome has a more progressive course in patients with lung cancer. AB - We studied whether a difference exists in the development of symptoms of the Lambert-Eaton myasthenic syndrome (LEMS) between patients with or without small cell lung cancer (SCLC). We assessed symptoms in 38 LEMS patients, 13 with SCLC, by interviewing them using a structured checklist, backed up by a review of their clinical records, and compared the frequency and time scale of symptoms during the course of LEMS. Bulbar (87%) and autonomic (95%) symptoms for the whole group were more common than reported in the literature. Frequencies of symptoms did not differ significantly between patients with and without SCLC, but symptoms in patients with SCLC appeared within a shorter time-frame, indicating a more rapid clinical course. The presence of a particular symptom associated with LEMS did not predict the presence of SCLC, but in patients with rapidly progressive LEMS the possibility of underlying lung cancer should be of particular concern. PMID- 15793846 TI - Spinal and cranial hypertrophic neuropathy in multiple sclerosis. AB - Two patients with multiple sclerosis developed symptomatic chronic inflammatory demyelinating polyneuropathy with massive spinal or cranial nerve hypertrophy revealed by neuroimaging. Sural nerve biopsy in one showed only moderate demyelination, axonal loss, and onion-bulb formation, illustrating dichotomy between severe proximal and milder distal nerve involvement. Patients with coexistent central and peripheral demyelination usually are symptomatic from dysfunction at one site or the other, but not from both. Our patients showed minimal response to steroids, intravenous immunoglobulin, or azathioprine. These cases suggest that the mechanism of disease in symptomatic central and peripheral demyelination may differ from that of disease in only one region, and that optimal therapy in this situation must be explored further. PMID- 15793847 TI - Concanavalin A inhibits pathophysiological effects of anti-ganglioside GQ1b antibodies at the mouse neuromuscular synapse. AB - Anti-GQ1b antibodies are present in the Miller Fisher syndrome (MFS), a monophasic neuropathy characterized by ataxia, areflexia, ophthalmoplegia, and sometimes cranial muscle weakness. We have previously shown, at the mouse neuromuscular junction (NMJ) ex vivo, that anti-GQ1b antibodies, through complement classic pathway activation, block synaptic transmission in a way that resembles the effect of the pore-forming alpha-latrotoxin (alphaLTx). In order to clarify the mechanism of these alphaLTx-like effects, including possible involvement of the alternative and mannose-binding protein complement pathways, we studied the effects of concanavalin A (ConA), a lectin known to block the action of alphaLTx, immunoglobulins, and early complement components. With electrophysiological, immunohistological, and bioassay experiments, we showed that the alphaLTx-like effects of anti-GQ1b antibody and complement were inhibited by pre- and coincubation with ConA. However, ConA was not able to inhibit evolution of alphaLTx-like effects when coincubated upon addition of complement at NMJs that had already bound anti-GQ1b antibody. Our data suggest that the mannose-binding protein pathway is not involved in the alphaLTx-like effect and that the inhibiting effect of ConA principally arises through interference with presynaptic binding of anti-GQ1b antibody. In control experiments, ConA prevented the neuroexocytotic effects of alphaLTx, indicating that alphaLTx receptors were inhibited under these conditions. We conclude that, although the physiological effects at the NMJ of anti-GQ1b antibody and alphaLTx are very similar, the activity of anti-GQ1b antibody is not mediated through activation of alphaLTx receptors, but rather is caused by direct presynaptic membrane damage through classic complement pathway activation. PMID- 15793848 TI - Fibrosis correlates with a ductular reaction in hepatitis C: roles of impaired replication, progenitor cells and steatosis. AB - The mechanisms for progressive fibrosis and exacerbation by steatosis in patients with chronic hepatitis C (HCV) are still unknown. We hypothesized that proliferative blockade in HCV-infected and steatotic hepatocytes results in the default activation of hepatic progenitor cells (HPC), capable of differentiating into both biliary and hepatocyte lineages, and that the resultant ductular reaction promotes portal fibrosis. To study this concept, 115 liver biopsy specimens from subjects with HCV were scored for steatosis, inflammation, and fibrosis. Biliary epithelium and HPC were decorated by cytokeratin 7 immunoperoxidase, and the replicative state of hepatocytes was assessed by p21 and Ki-67 immunohistochemistry. A ductular reaction at the portal interface was common. There was a highly significant correlation between the area of ductular reaction and fibrosis stage (r = 0.453, P < .0001), which remained independently associated after multivariate analysis. HPC numbers also correlated with fibrosis (r = 0.544, P < .0001) and the ductular area (r = 0.624, P < .0001). Moreover, steatosis correlated with greater HPC proliferation (r = 0.372, P = .0004) and ductular reaction (r = 0.374, P < .0001) but was not an obligate feature. Impaired hepatocyte replication by p21 expression was independently associated with HPC expansion (P = .002) and increased with the body mass index (P < .001) and lobular inflammation (P = .005). In conclusion, the strong correlation between portal fibrosis and a periportal ductular reaction with HPC expansion, the exacerbation by steatosis, and the associations with impaired hepatocyte replication suggest that an altered regeneration pathway drives the ductular reaction. We believe this triggers fibrosis at the portal tract interface. This may be a stereotyped response of importance in other chronic liver diseases. PMID- 15793849 TI - Late liver-related mortality from complications of transfusion-acquired hepatitis C. AB - Although several cohort studies have been reported in individuals with chronic hepatitis C virus (HCV) infection, little is known about liver-related mortality among the elderly. We conducted a cohort study in 302 patients with tuberculosis sequelae who had received a blood transfusion at a young age and had subsequently been treated at a chest clinic. The cohort consisted of 147 patients with antibody to HCV (anti-HCV), of whom 81% were positive for HCV RNA, and 155 without anti-HCV. The cohort was followed for a mean duration of 5.7 years. There were no differences between the two groups in the mean age of the patients at the time of transfusion (31 vs. 34 years) or at the time of entry into the study (65 vs. 66 years). The outcome of 143 patients with, and 145 without, anti-HCV could be traced; 92 (64%) and 82 (57%) had died, respectively. The main cause of death was tuberculosis sequelae in 61 (42%) and 66 (46%) patients, respectively. Eight (6%) of the 143 patients with anti-HCV died of liver disease (hepatocellular carcinoma: seven; rupture of varices: one). The average annual mortality from liver disease from study entry in the patients with anti-HCV was 9.8 per 1,000 person-years. The patients with anti-HCV had a significantly lower cause-specific survival probability for liver disease (92% vs. 100% at 10 years, P < .005). In conclusion, in our study, liver-related mortality appeared to be high among elderly HCV-infected individuals. PMID- 15793850 TI - Suppressive effect of ursodeoxycholic acid on type IIA phospholipase A2 expression in HepG2 cells. AB - Phospholipase A(2) IIA (PLA(2)IIA), which plays a crucial role in arachidonic acid metabolism and in inflammation, is upregulated under various pathological conditions, including in the gallbladder and gallbladder bile from patients with multiple cholesterol gallstones, in the liver and kidney of rats with cirrhosis, as well as in the colonic tissue of animals treated with a chemical carcinogen. The administration of ursodeoxycholic acid (UDCA) partially attenuated the PLA(2)IIA expression level in these different models. The aim of this study was to investigate the modulatory effect of UDCA on the PLA(2)IIA expression level at the cellular level. The HepG2 cells were selected to investigate the direct inhibitory effect of UDCA on PLA(2)IIA expression level. The proinflammatory cytokines (interleukin-6 and tumor necrosis factor alpha) -induced PLA(2)IIA expression in HepG2 cells was partially inhibited by the presence of UDCA in a dose-dependent fashion. The effect of UDCA on proinflammatory cytokines-induced PLA(2)IIA expression occurred at the transcriptional level. In addition, among the bile acids tested, this inhibitory effect was UDCA-specific. In conclusion, this study supports the possible alteration of arachidonic acid metabolism and PLA(2)IIA expression level, in particular, as the protective action of UDCA in patients with chronic liver disease. PMID- 15793851 TI - Small gallstones, preserved gallbladder motility, and fast crystallization are associated with pancreatitis. AB - Acute pancreatitis is a severe complication of gallstones with considerable mortality. We sought to explore the potential risk factors for biliary pancreatitis. We compared postprandial gallbladder motility (via ultrasonography) and, after subsequent cholecystectomy, numbers, sizes, and types of gallstones; gallbladder bile composition; and cholesterol crystallization in 21 gallstone patients with previous pancreatitis and 30 patients with uncomplicated symptomatic gallstones. Gallbladder motility was stronger in pancreatitis patients than in patients with uncomplicated symptomatic gallstones (minimum postprandial gallbladder volumes: 5.8 +/- 1.0 vs. 8.1 +/- 0.7 mL; P = .005). Pancreatitis patients had more often sludge (41% vs. 13%; P = .03) and smaller and more gallstones than patients with symptomatic gallstones (smallest stone diameters: 2 +/- 1 vs. 8 +/- 2 mm; P = .001). Also, crystallization occurred much faster in the bile of pancreatitis patients (1.0 +/- 0.0 vs. 2.5 +/- 0.4 days; P < .001), possibly because of higher mucin concentrations (3.3 +/- 1.9 vs. 0.8 +/- 0.2 mg/mL; P = .04). No significant differences were found in types of gallstones, relative biliary lipid contents, cholesterol saturation indexes, bile salt species composition, phospholipid classes, total protein or immunoglobulin (G, M, and A), haptoglobin, and alpha-1 acid glycoprotein concentrations. In conclusion, patients with small gallbladder stones and/or preserved gallbladder motility are at increased risk of pancreatitis. The potential benefit of prophylactic cholecystectomy in this patient category has yet to be explored. PMID- 15793853 TI - Prevalence and significance of neurocognitive dysfunction in hepatitis C in the absence of correlated risk factors. AB - Neurocognitive morbidity has been reported in individuals with chronic hepatitis C virus (HCV) infection, but the magnitude of such dysfunction in the absence of disease-correlated factors known to affect the central nervous system (e.g., substance abuse, cirrhosis, depression, interferon treatment) and the impact of any such change on functioning is unclear. We investigated a cohort of individuals with HCV, all of whom were carefully screened to exclude relevant comorbidities, to elucidate virus-related changes in the brain using neuropsychological tests and magnetic resonance spectroscopy (MRS). A cohort of 37 patients with chronic HCV infection was culled from 300 consecutive patients presenting to a tertiary care liver clinic. A comparison group of healthy controls (n = 46) was also assessed. Of 10 neurocognitive measures evaluated, the HCV group showed marginally poorer learning efficiency compared with controls; only 13% of patients demonstrated a clinical level of impairment on this test (defined as 1.5 SD below the normative standard). Although patients reported greater levels of fatigue and symptoms of depression, these factors did not correlate with the degree of learning inefficiency. With respect to MRS, the HCV group demonstrated increased choline and reduced N-acetyl aspartate relative to controls in the central white matter. Indicators of liver disease severity did not correlate with either memory or MRS abnormalities. In conclusion, while our findings support an association between hepatitis C and indicators of central nervous system involvement in a cohort of patients carefully screened to eliminate other factors influencing neurocognitive integrity, the clinical significance of these effects is limited. PMID- 15793852 TI - A biphasic response of hepatobiliary cholesterol metabolism to dietary fat at the onset of obesity in the mouse. AB - Human obesity is associated with abnormal hepatic cholesterol homeostasis and resistance to leptin action. Because leptin administration to rodents promotes the biliary elimination of plasma cholesterol, this study was designed to elucidate a pathophysiological role for leptin during the development of obesity. We fed mice diets containing high or low saturated fat contents. Before and after the onset of obesity, we measured downstream targets of leptin action and evaluated plasma, hepatic, and biliary cholesterol metabolism. Although not obese at 28 days, mice fed a high fat diet became hyperleptinemic. Sensitivity to leptin was evidenced by downregulation of both hepatic stearoyl CoA desaturase-1 and fatty acid synthase. Due principally to upregulation of adenosine triphosphate-binding cassette proteins A1 and G5, plasma high density lipoprotein (HDL) cholesterol concentrations increased, as did relative secretion rates of biliary cholesterol. A smaller, more hydrophilic bile salt pool decreased intestinal cholesterol absorption. In this setting, hepatic cholesterol synthesis was downregulated, indicative of increased uptake of plasma cholesterol. After 56 days of high fat feeding, obesity was associated with leptin resistance, as evidenced by marked hyperleptinemia without downregulation of stearoyl CoA desaturase-1 or fatty acid synthase and by upregulation of hepatic cholesterol and bile salt synthesis. Hypercholesterolemia was attributable to overproduction and decreased clearance of large HDL(1) particles. In conclusion, before the onset of obesity, preserved leptin sensitivity promotes biliary elimination of endogenous cholesterol in response to dietary fat. Leptin resistance due to obesity leads to a maladaptive response whereby newly synthesized cholesterol in the liver is eliminated via bile. PMID- 15793855 TI - Adiponectin as a protective factor in hepatic steatosis. AB - AIM: Obesity and insulin resistance (IR) are closely related to hepatic steatosis (HS), and adiponectin is a hepatic insulin sensitizer that has important effects in liver function. This study aims at investigating the relationship between serum adiponectin concentration and the presence of HS. METHODS: We carried out a cross-sectional study in a check-up unit of a University Hospital in Mexico City. We enrolled 196 subjects, comprising 98 subjects with HS (27 women, 71 men) and 98 controls (37 women and 61 men). Anthropometric, metabolic and biochemical variables were measured in the two groups. Serum adiponectin and leptin concentrations were determined, their association with grade of HS tested, and concentrations, according to quartiles, compared between cases and controls. chi(2) analysis for linear trends was used to test for a dose-response relationship and logistic regression analysis was conducted to test for a protective effect of adiponectin. RESULTS: The HS subjects were older and more obese than controls, with a central obesity pattern. In the fourth quartile of adiponectin concentrations, HS was less common and severe. In a multivariate model of the fourth quartile of the adiponectin concentrations, we observed a protective effect (OR = 0.17, 95%CI: 0.04-0.67, P = 0.01). In subjects with more severe HS, we observed higher leptin concentrations, and caloric intakes, total fat and iron consumption were higher than in controls. CONCLUSION: The results of the present study suggest that a high serum concentration of adiponectin is associated with a protective effect against HS. PMID- 15793854 TI - Molecular physiology of vesicular glutamate transporters in the digestive system. AB - Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system (CNS). Packaging and storage of glutamate into glutamatergic neuronal vesicles require ATP-dependent vesicular glutamate uptake systems, which utilize the electrochemical proton gradient as a driving force. Three vesicular glutamate transporters (VGLUT1-3) have been recently identified from neuronal tissue where they play a key role to maintain the vesicular glutamate level. Recently, it has been demonstrated that glutamate signaling is also functional in peripheral neuronal and non-neuronal tissues, and occurs in sites of pituitary, adrenal, pineal glands, bone, GI tract, pancreas, skin, and testis. The glutamate receptors and VGLUTs in digestive system have been found in both neuronal and endocrinal cells. The glutamate signaling in the digestive system may have significant relevance to diabetes and GI tract motility disorders. This review will focus on the most recent update of molecular physiology of digestive VGLUTs. PMID- 15793856 TI - Effects of glutamine supplementation on splenocyte cytokine mRNA expression in rats with septic peritonitis. AB - AIM: To investigate the effects of glutamine (GLN)-enriched diets before and GLN containing total parenteral nutrition (TPN) after sepsis or both on the secretion of cytokines and their mRNA expression levels in splenocytes of rats with septic peritonitis. METHODS: Rats were assigned to a control group and 4 experimental groups. The control group and experimental groups 1 and 2 were fed a semipurified diet, while experimental groups 3 and 4 had part of the casein replaced by GLN which provided 25% of the total nitrogen. After rats were fed with these diets for 10 d, sepsis was induced by cecal ligation and puncture (CLP), whereas the control group underwent a sham operation, at the same time, an internal jugular vein was cannulated. All rats were maintained on TPN for 3 d. The control group and experimental groups 1 and 3 were infused with conventional TPN, while the TPN in experimental groups 2 and 4 was supplemented with GLN, providing 25% of the total nitrogen in the TPN solution. All rats were kiued 3 d after sham operation or CLP to examine their splenocyte subpopulation distribution and cytokine expression levels. RESULTS: Most cytokines could not be detected in plasma except for IL-10. No difference in plasma IL-10 was observed among the 5 groups. The IL 2, IL-4, IL-10, and TNF-alpha mRNA expression levels in splenocytes were significantly higher in experimental groups 2 and 4 than in the control group and group 1. The mRNA expression of IFN-gamma was significantly higher in the GLN supplemented groups than in the control group and experimental group 1. The proportion of CD45Ra+ was increased, while those of CD3+ and CD4+ were decreased in experimental group 1 after CLP was performed. There were no differences in spleen CD3+ lymphocyte distributions between the control and GLN-supplemented groups. CONCLUSION: GLN supplementation can maintain T-lymphocyte populations in the spleen and significantly enhance the mRNA expression levels of Th1 and Th2 cytokines and TNF-alpha in the spleen of rats with septic peritonitis. PMID- 15793857 TI - Preventive and therapeutic effects of NF-kappaB inhibitor curcumin in rats colitis induced by trinitrobenzene sulfonic acid. AB - AIM: To ascertain the molecule mechanism of nuclear factor-kappaB (NF-kappaB) inhibitor curcumin preventive and therapeutic effects in rats' colitis induced by trinitrobenzene sulfonic acid (TNBS). METHODS: Sixty rats with TNBS-induced colitis were treated with 2.0% curcumin in the diet. Thirty positive control rats were treated with 0.5% sulfasalazine (SASP). Thirty negative control rats and thirty model rats were treated with general diet. Changes of body weight together with histological scores were evaluated. Survival rates were also evaluated. Cell nuclear NF-kappaB activity in colonic mucosa was evaluated by using electrophoretic mobility shift assay. Cytoplasmic IkappaB protein in colonic mucosa was detected by using Western Blot analysis. Cytokine messenger expression in colonic tissue was assessed by using semiquantitative reverse-transcription polymerase chain reaction. RESULTS: Treatment with curcumin could prevent and treat both wasting and histopathologic signs of rats with TNBS-induced intestinal inflammation. In accordance with these findings, NF-kappaB activation in colonic mucosa was suppressed in the curcumin-treated groups. Degradations of cytoplasmic IkappaB protein in colonic mucosa were blocked by curcumin treatment. Proinflammatory cytokine messenger RNA expression in colonic mucosa was also suppressed. CONCLUSION: This study shows that NF-kappaB inhibitor curcumin could prevent and improve experimental colitis in murine model with inflammatory bowel disease (IBD). The findings suggest that NF-kappaB inhibitor curcumin could be a potential target for the patients with IBD. PMID- 15793858 TI - Expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic stellate cells during rat hepatic fibrosis and its intervention by IL-10. AB - AIM: To investigate the expression of matrix metallopr-oteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic fibrosis and the antifibrogenic role of exogenous interleukin-10 (IL-10). METHODS: Hepatic fibrosis was induced by CCl(4) administration and 60 male Sprague-Dawley rats were randomly divided into normal control group (group N, 8 rats), CCl(4)-induced group (group C, 28 rats) and IL-10-treated group (group I, 24 rats). At the beginning of the 7(th) and 11(th) wk, rats in each group were routinely perfused with pronase E and type IV collagenase through portal vein catheter and the suspension was centrifuged by 11% Nycodenz density gradient to isolate hepatic stellate cells (HSCs). RT-PCR was used to analyze mRNA of MMP-2 and TIMP-1 from freshly isolated cells. Densitometric data were standardized with beta-actin signals. Immunocytochemistry was performed to detect MMP-2 and TIMP-1 expression in HSC cultured for 72 h. RESULTS: Compared to group N in the 7(th) wk, MMP-2 and TIMP-1 mRNA increased in group C (P = 0.001/0.001) and group I (P = 0.001/0.009). The level of MMP-2 and TIMP-1 mRNA in group I was significantly lower than that in group C (P = 0.001/0.001). In the 11(th) wk, MMP-2 mRNA in group I was still lower than that in group C (P = 0.005), but both dropped compared with that in the 7(th) week (P = 0.001/0.004). TIMP-1 mRNA in group I was still lower than that in group C (P = 0.001), and increased in group C (P = 0.001) while decreased in group I (P = 0.042) compared with that in the 7(th) wk. Same results were found by immunocytochemistry. CONCLUSION: Expression of MMP-2 and TIMP-1 is increased in hepatic fibrosis. IL-10 exhibits an antifibrogenic effect by suppressing MMP-2 and TIMP-1 expression. PMID- 15793860 TI - Protective effects of 5,4'-dihydroxy-3',5'-dimethoxy-7-O-beta-D glucopyranosyloxy-flavone on experimental hepatic injury. AB - AIM: To investigate the pharmacological effects of rice flavone (5,4'-dihydroxy 3',5'-dimethoxy-7-O-beta-D-glucopyranosyloxy-flavone, RF) separated from panicle differentiating to flowing rice on rat experimental hepatic injury. METHODS: Models of rat acute hepatic injury induced by carbon tetrachloride (CCl(4)) administration, rat hepatic fibrosis induced by thioacetamide, injury of primary cultured rat hepatocytes induced by CCl(4), respectively, were established. After treated with RF, content of serum alanine transaminase (ALT), aspartate aminotransferase (AST) and albumin (Alb), hyaluronic acid (HA), the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and hydroxyproline (Hyp) were measured and liver tissue was observed pathologically by hematoxylin eosin (HE) staining. Effects of RF on pathological changes, function index, enzyme of scavenging free radicals and blood rheology were evaluated. RESULTS: In model of rat acute hepatic injury induced by CCl(4), RF can significantly decrease the contents of serum ALT, AST, increase the content of Alb, improve the dropsy and fat denaturalization of hepatocytes. In model of rat hepatic fibrosis induced by thioacetamide, RF can inhibit the increase of HA, Hyp and whole blood viscosity, and improve the activities of GSH-Px and SOD, and inauricular microcirculation. CONCLUSION: RF has apparent protective effects on hepatic injury by increasing activity of GSH-Px and SOD, scavenging free radicals produced by CCl(4), reducing blood viscosity, and improving microcirculation and blood supply. PMID- 15793859 TI - Expression and significance of nuclear factor kappaB p65 in colon tissues of rats with TNBS-induced colitis. AB - AIM: To investigate the role of NF-kappaB in the pathogenesis of TNBS-induced colitis in rats. METHODS: Thirty-two healthy adult Sprague-Dawley (SD) rats were randomly divided into four groups of eight each: normal, NS, model I, model II groups in our study. Rat colitis model was established through 2-,4-,6 trinitrobenzene sulfonic acid (TNBS) enema. At the end of four weeks, the macroscopical and histological changes of the colon were examined and mucosa myeloperoxidase (MPO) activities assayed. NF-kappaB p65 expression was determined by Western blot assessment in cytoplasmic and nuclear extracts of colon tissue, and the expressions of TNF-alpha and ICAM-1 protein in colon tissue were examined by immunohistochemistry. The relativities between expression of NF-kappaB p65 and other parameters were analyzed. RESULTS: TNBS enema resulted in pronounced pathological changes of colonic mucosa in model II group (macroscopic and histological injury indices 6.25+/-1.39 and 6.24+/-1.04, respectively), which were in accordance with the significantly elevated MPO activity (1.69+/-0.11). And the nuclear level of NF-kappaB and expression of TNF-alpha, ICAM-1 in rats of model II group were higher than that of normal control (9.7+/-1.96 vs 1.7+/-0.15, 84.09+/-14.52 vs 16.03+/-6.21, 77.69+/-8.09 vs 13.41+/-4.91 P<0.01), Linear correlation analysis revealed that there were strong correlations between the nuclear level of NF-kappaB and the tissue positive expression of TNF-alpha and ICAM-1, MPO activities, macroscopical and histological indices in TNBS-induced colitis, respectively (r = 0.8235, 0.8780, 0.8572, 0.9152, 0.8247; P<0.05). CONCLUSION: NF-kappaB plays a pivotal role in the pathogenesis of ulcerative colitis, which might account for the up-regulation the expression of TNF-alpha and ICAM-1. PMID- 15793861 TI - Psychiatric side effects of pegylated interferon alfa-2b as compared to conventional interferon alfa-2b in patients with chronic hepatitis C. AB - AIM: To assess systematically the spectrum and extent of depressive symptoms comparing patient groups receiving peginterferon or conventional interferon. METHODS: Ninety-eight patients with chronic hepatitis C and interferon-based therapy (+ribavirin) were consecutively enrolled in a longitudinal study. Patients were treated with conventional interferon alfa-2b (48/98 patients; 5 MIU interferon alfa-2b thrice weekly) or peginterferon alfa-2b (50/98 patients; 80 150 mug peginterferon alfa-2b) in combination with weight-adapted ribavirin (800 1 200 mg/d). Repeated psychometric testing was performed before, three times during and once after antiviral therapy: Depression was evaluated by the Hospital Anxiety and Depression Scale (HADS), anger/hostility by the Symptom Checklist-90 Items Revised (SCL-90-R). RESULTS: Therapy with pegylated interferon alfa-2b produces comparable scores for depression (ANOVA: P = 0.875) as compared to conventional interferon. Maximums of depression scores were even higher and cases of clinically relevant depression were frequent during therapy with peginterferon. Scores for anger/hostility were comparable for both therapy subgroups. CONCLUSION: Our findings suggest that the extent and frequency of depressive symptoms in total are not reduced by peginterferon. Monitoring and management of neuropsychiatric toxicity especially depression have to be considered as much as in antiviral therapy with unmodified interferon. PMID- 15793862 TI - Significance of glucocorticoid receptor expression in colonic mucosal cells of patients with ulcerative colitis. AB - AIM: Glucocorticoid (GC) resistant ulcerative colitis (UC) remains a serious disease and is difficult to manage. Although the molecular basis of GC insensitivity is still unknown, GC receptors (GRalpha and GRbeta) may play an important role in it. This study was aimed to investigate the relationship between the expression of GRalpha and GRbeta in colonic mucosal cells of patients with UC, the efficacy of GC therapy and the intensity of inflammation. METHODS: Twenty-five cases of UC were classified into: GC sensitive (n = 16) and GC resistant (n = 9) cases. Patients consisted of mild (n = 6), moderate (n = 8) and severe (n = 11) cases. GRalpha and GRbeta expression in colonic mucosal specimens were investigated by immunohistochemistry, and compared between GC resistant and sensitive groups, and also among various degrees of inflammation. RESULTS: All cases were positive for GRalpha and GRbeta expression. Both positive association between GRalpha expression and the response of UC to GC and strong negative association between GRbeta expression and the response of UC to GC were identified. There was no significant association between GRalpha/GRbeta expression and the degree of inflammation of UC. CONCLUSION: These findings suggest that both GRalpha and GRbeta may play an important role in the action of GC, and that GRbeta functions as a dominant negative inhibitor of GRalpha. Expression of GRalpha and GRbeta in colonic mucosal cells of patients with UC may serve as predictors of glucocorticoid response, but can not function as markers of inflammatory intensity. PMID- 15793863 TI - Prognostic analysis of surgical treatment of peripheral cholangiocarcinoma: two decades of experience at Chang Gung Memorial Hospital. AB - AIM: To analyze the prognostic factors influencing the overall survival of peripheral cholangiocarcinoma (PCC) patients undergoing surgical treatment during 25 years at a single institution. METHODS: This study retrospectively reviewed prospectively collecting data about 373 patients with histologically proven PCC who underwent surgical treatment between 1977 and 2001. RESULTS: Three hundred and seventy-three PCC patients (159 men and 214 women) underwent surgical treatment from 1977 to 2001. Among them, 187 PCC patients underwent hepatectomy and 135 had curative resection (curative resectability rate: 36.2%). The follow up duration ranged from 1.05 to 167.6 mo (mean/median = 14.1/7.2 mo). Overall cumulative survival rates at 1, 3, and 5 years were 32.5%, 9.2%, and 4.1%, respectively. Univariate log-rank analysis identified the following as adverse influences on overall survival: presence of symptoms, absence of mucobilia, elevated CEA and CA 19-9 levels, non-papillary tumor type, receiving non hepatectomy, advanced tumor staging, lack of post-operative chemotherapy, and radiotherapy. Meanwhile, multivariate Cox's proportional hazard analysis demonstrated that absence of mucobilia, non-papillary tumor type, advanced tumor staging, non-hepatectomy, and lack of post-operative chemotherapy were the five independent prognostic factors that adversely affected overall survival. CONCLUSION: Favorable overall survival of PCC patients undergoing surgical treatment depends on early tumor stage, presence of mucobilia, papillary tumor type, hepatic resection, and post-operative chemotherapy. PMID- 15793864 TI - Incidence of ulcerative colitis in Central Greece: a prospective study. AB - AIM: To study the incidence of ulcerative colitis UC in the prefecture of Trikala, Central Greece. METHODS: A prospective and population based epidemiological study of UC from 1990 to the end of 1994 was conducted. Trikala is a semirural prefecture of Central Greece with a population of 138 946 (census 1991). Three gastroenterologists (one hospital based, two private doctors) of the prefecture participated in this study. RESULTS: During the study period, 66 new histologically verified cases of UC were recorded. The mean annual incidence of the disease in 1990-1994 was 11.2 per 10(5) inhabitants (95%CI: 8.7-14.3). There was no difference between men and women (annual incidence: 10.5 and 12.0 per 10(5) inhabitants respectively), either among urban, semirural or rural populations (annual incidence: 11.7, 17.1 and 9.9 per 10(5) inhabitants respectively). The majority (56%) of the patients never smoked and a quarter were ex-smokers. About a half of all cases had proctitis. CONCLUSION: UC is common in Central Greece and its incidence is similar to that in North-Western European countries. PMID- 15793865 TI - p62 Expression in primary carcinomas of the digestive system. AB - AIM: To characterize p62 expression and define the relationship between p62 expression and cell proliferation in primary carcinomas of the digestive system. METHODS: p62 expression was characterized in surgically resected tumor specimens from 60 patients with primary carcinomas of the digestive tract (including 22 esophageal carcinomas, 17 gastric carcinomas, and 21 colorectal carcinomas) and 40 patients with hepatocellular carcinoma (HCC) by immunohistochemistry (IHC). The cell proliferation was determined by IHC of Ki-67 in 40 patients with HCC. RESULTS: Twenty-two cases of esophageal carcinoma were histopathologically diagnosed as squamous cell carcinoma. We combined the gastric and colorectal carcinomas based on the equivalent histology. The 38 tumors in the combined groups, consisted of 17 well-differentiated, eight moderately differentiated, nine poorly differentiated carcinomas, and four mucinous adenocarcinomas. According to the criteria of Edmondson and Steiner, 40 patients with HCC were graded (2 grade I, 17 grade II and 21 grade III). p62 expression in primary carcinomas of the gastrointestinal tract (60/60,100%) was higher than that (27/40, 67.5%) of HCC (P<0.01, chi(2) = 19.63). High expression levels of p62 were positively correlated with histological grades in gastric and colorectal carcinomas (P<0.0001) and inversely associated with those in HCC (P = 0.0322). No significant correlations were observed for esophageal carcinomas (P = 0.8246). p62 expression was also detected in the cytoplasm of morphologically normal columnar epithelial cells adjacent to the cancer foci of gastric and colorectal carcinomas. In 40 HCC specimens, the mean Ki-67 labeling index (LI) was (19.6+/ 16.0)%. It was (28.3+/-18.73)% in 12 cases with high p62 expression (+++), (7.53+/-14.83)% in 13 cases without p62 expression(-). Patients with a high p62 expression showed a significantly higher level of Ki-67 staining than those without p62 expression (P<0.05, t = 2.069). CONCLUSION: p62 expression is common in carcinomas of the digestive system and higher in carcinomas of the gastrointestinal tract than in primary HCC. p62 is a cellular differentiation related protein. Cancer cells with a high p62 expression exhibited highgrowth fractions in HCC. PMID- 15793866 TI - Expression of CXC receptor 1 and 2 in esophageal mucosa of patients with reflux esophagitis. AB - AIM: Interleukin 8 (IL-8) mediates neutrophil trafficking via its receptors. Recent studies have shown that IL-8 is likely involved in the development and progression of erosive reflux esophagitis (RE), yet little is known about the two distinct receptors, CXC receptor (CXCR)-1 and -2. The purpose of this study was to determine CXCR-1 and -2 messenger RNA expression levels in RE. METHODS: We studied 26 patients with RE and 15 asymptomatic controls. Paired biopsy samples were taken from the esophagus 3 cm above the gastroesophageal junction; one biopsy was snap frozen for measurement of CXCR-1 and -2 mRNA levels by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and another was formalin-fixed for histopathological evaluation. We also examined the association of the expression levels of CXCR-1 and -2 mRNA with histopathological hallmarks of RE. RESULTS: The relative CXCR-1 and -2 mRNA expression levels were rather decreased in esophageal mucosa of patients with RE, compared to those in normal esophagus of controls. There were no significant difference in the relative mRNA expression levels of CXCR-1 and -2 among endoscopic grades of RE based on the Los Angeles classification. Each histopathological hallmark of GERD was not associated with the expression levels of CXCR-1 and -2 mRNA. CONCLUSION: Apart from overexpression of IL-8, the relative expression levels of CXCR-1 and 2 mRNA were rather lower than expected in the affected esophageal mucosa of patients with RE. PMID- 15793867 TI - Four-year follow up of hepatitis C patients vaccinated against hepatitis B virus. AB - AIM: Patients with chronic hepatitis C have been recommended to receive vaccinations against hepatitis B. Our study aimed at evaluating the hepatitis B immunogenicity and efficacy against hepatitis B virus infection 4 years after primary immunization series in a group of patients with chronic hepatitis C. METHODS: We recruited 36 out of 48 hepatitis C virus (HCV) infected individuals who were vaccinated against hepatitis B virus (20 mug of recombinant HBsAg at 0-1 6 mo schedule) in 1998. Here we measured anti-HBs titers and anti-HBc 4 years after delivery of the third dose of primary immunization series. RESULTS: After 4 years a total of 13/36 (36%) HCV infected patients had seroprotective titers of anti-HBs compared with 9/10 (90%) in the control group, (P<0.05). Similarly the mean concentration of anti-HBs found in hepatitis C patients was significantly lower than that found in healthy subjects (18.3 and 156.0 mIU/mL respectively (P<0.05). None of the HCV infected patients or controls became infected with HBV during the study period as confirmed by anti-HBc negativity. CONCLUSION: We demonstrated that 4 years after HBV immunizations' more than 60% of vaccinated HCV patients did not maintain seroprotective levels of anti-HBs, which might put them at risk of clinically significant breakthrough infections. Further follow-up studies are required to clarify whether memory B and T lymphocytes can provide protection in chronic hepatitis C patients in the absence or inadequate titers of anti-HBs. PMID- 15793869 TI - Therapeutic effect of autologous dendritic cell vaccine on patients with chronic hepatitis B: a clinical study. AB - AIM: To investigate the therapeutic effect of autologous HBsAg-loaded dendritic cells (DCs) on patients with chronic hepatitis B. METHODS: Monocytes were isolated from fresh peripheral blood of 19 chronic HBV-infected patients by Ficoll-Hypaque density gradient centrifugation and cultured by plastic-adherence methods. DCs were induced and proliferated in the culture medium with recombinant human granulocyte-macrophage-colony-stimulating factor (rhGM-CSF) and human interleukin-4 (rhIL-4). DCs pulsed with HBsAg for twelve hours were injected into patients subcutaneously twice at intervals of two weeks. Two patients received 100 mg oral lamivudine daily for 12 mo at the same time. HBV-DNA and viral markers in sera of patients were tested every two months. RESULTS: By the end of 2003, 11 of 19 (57.9%) patients had a clinical response to DC-treatment. HBeAg of 10 (52.6%) patients became negative, and the copies of HBV-DNA decreased 10(1.77+/-2.39) on average (t = 3.13, P<0.01). Two cases co-treated with DCs and lamivudine had a complete clinical response. There were no significant differences in the efficient rate between the cases with ALT level lower than 2XULN and those with ALT level higher than 2XULN before treatment (chi(2) = 0.0026). CONCLUSION: Autologous DC-vaccine induced in vitro can effectively suppress HBV replication, reduce the virus load in sera, eliminate HBeAg and promote HBeAg/anti-HBe transformation. Not only the patients with high serum ALT levels but also those with normal ALT levels can respond to DC vaccine treatment, and the treatment combining DCs with lamivudine can eliminate viruses more effectively. PMID- 15793868 TI - Deformity of duodenal bulb, gastric metaplasia of duodenal regenerating mucosa and recurrence of duodenal ulcer: a correlated study. AB - AIM: To investigate the correlation among the presence and degree of gastric metaplasia of duodenal regenerating mucosa, the deformity of bulb and the recurrence of duodenal ulcer. METHODS: A total of 99 patients with duodenal ulcer were treated with H(2)-antagonist with or without antimicrobial therapy. All patients received follow-up endoscopic examinations 6 wk after treatment. When the ulcer(s) were noted to be healed, two biopsies were taken from the ulcer scar for histological study of gastric metaplasia, and 4 biopsies were taken from antrum for Helicobacter pylori (H pylori) study. Out of these cases, 44 received further follow-up endoscopic examinations after 3, 6 and 12 mo respectively for studying the recurrence rate of duodenal ulcers. The correlation among ulcer recurrence, degree of gastric metaplasia of regenerating mucosa, bulbar deformity, and colonization of H pylori in the stomach was then studied. RESULTS: The results showed that there was a strong correlation between the deformity of duodenal bulb and the degree of gastric metaplasia of regenerating duodenal mucosa. The recurrence rate of duodenal ulcer had a significant difference between patients with and without H pylori colonization in the stomach (P<0.001). The greater the degree of gastric metaplasia of duodenal regenerating mucosa, the higher the recurrence rate of duodenal ulcer (P = 0.021). The more deformed the duodenal bulb, the higher the incidence of recurrence of duodenal ulcer (P = 0.03). CONCLUSION: There is a correlation among deformity of duodenal bulb, gastric metaplasia of duodenal regenerating mucosa and recurrence of duodenal ulcer. A more severely deformed duodenal bulb is closely related to a greater extent of gastric metaplasia. Both factors contribute to the recurrence of duodenal ulcer. PMID- 15793870 TI - Study on hepatoprotective effect of peptide S-8300 from shark liver. AB - AIM: To explore the effects of peptide S-8300 from shark liver (S-8300) on liver function as well as in regulating immune functions in experimental injury models. METHODS: Mice were administered with different doses of S-8300 for four consecutive days, followed by mice injected with tetrachloromethane (CCl(4)) on d 3. The activity of ALT, AST, LDH, SOD and contents of MDA and GSH in the mice liver were tested. And mice were treated with Cy (100 mg/kg) at the beginning of the experiment to induce immunosuppression model. The S-8300 groups were treated with S-8300 seven days after the beginning of Cy administration. The effects of S 8300 on the formulation of serum hemolysin and the content of IL-2 in serum in the immunosuppression mice were observed. RESULTS: S-8300 obviously decreased the level of ALT (52.2+/-11.0 U/dL vs 135.9+/-6.5 U/dL, P<0.01), AST (67.5+/-6.9 U/dL vs 238.8+/-8.7 U/dL, P<0.01), LDH (155.1+/-46.8 U/dL vs 240.4+/-6.0 U/dL, P<0.01) and MDA (0.64+/-0.027 nmol/mg vs 1.06+/-0.040 nmol/mg, P<0.01) and increased SOD (24.51+/-1.01 U/mg vs 19.05+/-0.73 U/mg, P<0.01) and GSH (24.17+/-0.91 microg/mg vs 14.93+/-0.45 microg/mg, P<0.01) in mice liver damaged by CCl(4). S-8300 also markedly improved the formulation of serum hemolysin (0.094+/-0.005 A(540) vs 0.063+/-0.006 A(540), P<0.01) and increased the level of IL-2 (9.74+/-1.16 pg/mL vs 5.81+/-0.87 pg/mL, P<0.01) in serum of the immunosuppression mice. CONCLUSION: The results suggested S-8300 has significant hepatoprotective, immunomodulatory and inhibiting of lipid peroxidation activity. PMID- 15793871 TI - Gastrointestinal bezoars: a retrospective analysis of 34 cases. AB - AIM: Bezoars (BZ) are the most common foreign bodies of gastrointestinal tract. Clinical manifestations vary depending on the location of BZ from no symptoms to acute abdominal syndrome. When located in small bowel, they frequently cause small bowel obstruction (SBO). We aimed to present our experience by reviewing literature. METHODS: Thirty-four patients with gastrointestinal BZ were presented. The data were collected from hospital records and analyzed retrospectively. Morbidity and mortality rates were statistically analyzed between the subgroups according to SBO and endoscopic or surgical treatment modalities. RESULTS: The 34 patients had phytobezoars (PBZ). Two patients with mental retardation and trichotillomania had trichobezoars (TBZ). More than half of them (55.88%) had previous gastric surgery. Also most of them had small bowel bezoars resulting in obstruction. Surgical and endoscopic morbidity rates were 32.14% and 14.28% respectively. The total morbidity rate of this study was 29.41%. Four patients in surgically treated group died. There was no death in endoscopically treated group. The total and surgical mortality rates were 11.76% and 14.28% respectively. The differences in morbidity and mortality rates between the subgroups were not statistically significant. CONCLUSION: BZ are commonly seen in stomach and small intestine. SBO is the most common complication. When uncomplicated, endoscopic or surgical removal can be applied easily. PMID- 15793872 TI - Age scope of high-risk population for esophageal cancer in Ci county. AB - AIM: To define the age scope of high-risk population for esophageal cancer (EC) in Ci county. METHODS: The results of endoscopic examination of 2 013 subjects, cytological screening of 16 763 persons and records of 9 265 patients with EC were analyzed by Ridit methods, the standard age group was 45-49 year group. RESULTS: The average age of patients with moderate esophageal epithelium dysplasia by endoscopic examination was 53.5 years, of severe esophageal epithelium dysplasia, 51.4 years, early EC, 55.6 years. The average age of stage one severe epithelium dysplasia (SEEDI) by cytological screening was 51.2 years, of stage two severe epithelium esophageal dysplasia (SEED II) 51.6 years, of advanced EC 61.7 years. In the group of 40-year olds, the value of Ridit by pathological diagnosis was 0.46, 95% CI, 0.45-0.47, that by cytological diagnosis was 0.45, 95% CI, 0.43-0.47. As the age increased at five-year intervals, the value of Ridit increased significantly. CONCLUSION: In Ci county of a high incidence area of EC, the age definition of high-risk population should be above 45 years. PMID- 15793873 TI - Stereoselective urinary excretion of S-(-)- and R-(+)-propranolol glucuronide following oral administration of RS-propranolol in Chinese Han subjects. AB - AIM: To study the stereoselectivity of phase II glucuronidation metabolism of side-chain propranolol in Chinese Han population. METHODS: Sixteen adult Chinese Han volunteers with an average age of 20 years were given a single oral dose of 20 mg racemic propranolol. Human urine at indicated time after administration was collected and S-(-)-propranolol glucuronide and R-(+)-propranolol glucuronide were determined simultaneously by using RP-HPLC. RESULTS: The mean values of k were 0.19+/-0.04 h(-1) and 0.28+/-0.06 h(-1), of t(1/2) 3.56+/-0.73 h and 2.45+/ 0.50 h, of T(max) 2.21+/-0.45 and 1.75+/-0.33 h, and of Xu(0-24) 5.65+/-0.98 and 2.95+/-0.62 micromoL for S-(-)- and R-(+)-propranolol glucuronide, respectively. The cumulative excretion percentages in urine of doses were 14.7+/-2.46% and 7.68+/-1.60% for S-(-)- and R-(+)-propranolol glucuronide, respectively. The results showed the elimination rate constant k of S-(-)-propranolol glucuronide was less than that of R-(+)-propranolol glucuronide; and the elimination half life (t(1/2)), T(max) and the cumulative excretion amount(Xu(0-24)) of R-(+) propranolol glucuronide were significantly less than that of S-(-)-propranolol glucuronide. CONCLUSION: The propranolol glucuronidation of the side-chain undergoes stereoselective excretion in Chinese Han population after an oral administration of racemic propranolol. PMID- 15793875 TI - Apoptosis of human gastric cancer SGC-7901 cells induced by mitomycin combined with sulindac. AB - AIM: To investigate the effects of mitomycin (MMC) combined with sulindac on cell viability, apoptotic induction and expression of apoptosis-related gene Bcl-2 and cyclooxygenase-2 (COX-2) in gastric cancer SGC-7901 cells. METHODS: Human gastric cancer SGC-7901 cells were divided into three treatment groups,namely sulindac treatment group, MMC treatment group and combined sulindac with MMC treatment group. After being treated with drugs, cell viability was examined by MTT assay. Flow cytometry was used to evaluate the cell cycle distribution and apoptotic rates. Morphology of the cells was observed under light microscope and interactive laser microscope. Expression of COX-2 and Bcl-2 was determined by immunocytochemical method. RESULTS: After exposure for 12 h to three kinds of drugs, gastric cancer SGC-7901 cells presented some morphological features of apoptosis, including cell shrinkage, nuclear condensation, DNA fragmentation and formation of apoptotic bodies. Growth inhibition was more obvious in combined sulindac with MMC treatment group and sulindac treatment group than in MMC treatment group. The apoptotic rates in co-treated cells and MMC-treated cells 24 h after treatment were 12.0% and 7.2%, respectively. After exposure for 24 h to MMC, the expression of COX-2 and Bcl-2 protein was up-regulated, COX-2 levels were down-regulated but Bcl-2 gene expression was not changed significantly in combined treatment group. CONCLUSION: MMC-induced apoptosis is reduced by up regulating the expression of COX-2 and Bcl-2 genes. MMC combined with sulindac can suppress the growth of gastric cancer cells through induction of apoptosis mediated by down-regulation of apoptosis-related Bcl-2 and COX-2 gene. PMID- 15793874 TI - Modulation of liver oxidant-antioxidant system by ischemic preconditioning during ischemia/reperfusion injury in rats. AB - AIM: To investigate effects of ischemic pre-conditioning on the liver endogenous oxidant-antioxidant system during ischemia/reperfusion injury. METHODS: Twenty four male Sprague-Dawley rats were randomly divided into sham-operated (Sham), ischemia/reperfusion (I/R), ischemic pre-conditioning plus ischemia/reperfusion (IPC) groups. Serum ALT, AST and hyaluronic acid levels were assayed and pathologic alterations observed. Liver malondialdehyde (MDA) contents, endogenous antioxidant enzymes, superoxidase dismutase (SOD), catalase (CAT), gultathionine peroxidase (GSH-Px) activities, neutrophils accumulation marker, myeloperoxidase (MPO) activities were measured respectively. RESULTS: Compared with I/R group, sinusoidal endothelial cells as well as hepatocytes damages, as assessed biochemically and histochemically, were improved significantly in IPC group; neutrophils infiltration was also markedly reduced. In IPC group, liver peroxidation, as measured by MDA contents, was significantly decreased when compared with I/R group; endogenous antioxidant enzymes, SOD, CAT and GSH-Px activities were markedly higher than that in I/R group. CONCLUSION: Ischemic pre conditioning exerts protective effects on both hepatic sinusoidal endothelial cells and hepatocytes during liver I/R injury. Its mechanisms may involve dimunition of neutrophils infiltration and modulation of the imbalance of endogenous oxidant-antioxidant system in the organism. PMID- 15793876 TI - Oral attenuated Salmonella typhimurium vaccine against MG7-Ag mimotope of gastric cancer. AB - AIM: To develop an oral attenuated Salmonella typhimurium vaccine against gastric cancer and to evaluate its efficacy in mice. METHODS: A complementary sequence of Nco I site and a sequence coding for MG7-Ag mimotope were designed at the 5' terminus of forward primer. Using p1.2 II-HBCAg plasmid as template, PCR was performed to get a fusion gene of the mimotope and a HBcAg gene. The fusion gene was then subcloned into the plasmid pYA3341 complementary to Salmonella typhimurium X4550, and the recombinant plasmid was then transformed into attenuated Salmonella typhimurium X4550. Balb/c mice were orally immunized with the recombinant Salmonella typhimurium X4550. The mice were immunized every 2 wk to reinforce the immunity. At the 6th wk, serum titer of antibody was detected by ELISA, and at the 8th wk, cellular immunity was detected by (51)Cr release test. Ehrlich ascites carcinoma cells expressing MG7-Ag were used in tumor challenge assay as a model to evaluate the protective effect of the vaccine. RESULTS: Serum titer of antibody against MG7-Ag was significantly higher in mice immunized with the vaccine than in control groups (0.9538+/-0.043 vs 0.6531+/-0.018, P<0.01; 0.9538+/-0.043 vs 0.6915+/-0.012, P<0.01), while in vitro (51)Cr release assay of the splenocytes showed no statistical difference in the three groups. Two weeks after tumor challenge, 1 in 5 immunized mice was tumor free, while all the mice in the control group presented tumor. CONCLUSION: Oral attenuated Salmonella typhimurium vaccine against the MG7-Ag mimotope of gastric cancer is immunogenic. It can induce significant humoral immunity against tumors in mice, and has some protective effects. PMID- 15793878 TI - Reduced bone mineral density and altered bone turnover markers in patients with non-cirrhotic chronic hepatitis B or C infection. AB - AIM: Previous studies suggest that loss of bone mineral density (BMD) frequently occurs in patients with chronic viral liver disease, presenting with histologically proven liver cirrhosis. However, little is known about the occurrence of bone disease in non-cirrhotic patients with chronic hepatitis B or C. Therefore, it was the aim of this study to evaluate this particular population for BMD and bone turnover markers. METHODS: Biochemical markers of bone turnover and BMD were measured in 43 consecutive patients with HCV (n = 30) or HBV (n = 13) infection without histological evidence for liver cirrhosis. Mean age was 49 years (range 26-77 years). BMD was measured by dual X-ray absorptiometry in the femoral neck (FN) and the lumbar spine (LS) region. In addition, bone metabolism markers were measured. RESULTS: BMD was lowered in 25 (58%) of the patients with chronic hepatitis B or C (FN: 0.76 (0.53-0.99); LS: 0.96 (0.62-1.23) g/cm(2)). Eight (32%) osteopenic patients were diagnosed with osteoporosis. Bone-specific alkaline phosphatase (P = 0.005) and intact parathyroid hormone (iPTH) (P = 0.001) were significantly elevated in the more advanced stages of fibrosis. Mean T-score value was lower in patients with chronic hepatitis C as compared to patients suffering from chronic hepatitis B; however, the difference was not statistically significant (P = 0.09). CONCLUSION: There was a significantly reduced BMD in non-cirrhotic patients with chronic hepatitis B or C infection. Alterations of bone metabolism already occurred in advanced liver fibrosis without cirrhosis. According to our results, these secondary effects of chronic viral hepatitis should be further investigated. PMID- 15793877 TI - Screening differentially expressed genes in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis. AB - AIM: To screen genes differentially expressed in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis. METHODS: A subtracted cDNA library of mouse hepatocarcinoma cell line with high potential of lymphatic metastatic Hca-F and its synogenetic cell line Hca-P with a low metastatic potential was constructed by suppression subtracted hybridization(SSH) method. The screened clones of the subtracted library were sequenced and GeneBank homology search was performed. RESULTS: Fourteen differentially expressed cDNA fragments of Hca-F were obtained with two novel genes. CONCLUSION: SSH is a useful technique to detect differentially expressioned genes and an effective method to clone novel genes. PMID- 15793879 TI - Hepatic steatosis is associated with an increased risk of carotid atherosclerosis. AB - AIM: Although an association between hepatic steatosis and vascular risk factors has been described, direct relationships between fatty liver and atherosclerosis have not yet been investigated. The aim of the present study has been to investigate those relationships. METHODS: The Study of Health in Pomerania examined a random population sample aged between 20 and 79 years. A study population of 4 222 subjects without hepatitis B and C infections and without liver cirrhosis was available for the present analysis. Hepatic steatosis was defined sonographically and intima-media thickness (IMT) as well as plaque prevalence were estimated by carotid ultrasound. RESULTS: The prevalence rate of hepatic steatosis was 29.9%. Among subjects aged > or =45 years, an association between hepatic steatosis and IMT of the carotid arteries was found in bivariate analysis, but not after adjustment for atherosclerotic risk factors. Individuals with fatty liver had more often carotid plaques than persons without fatty liver (plaque prevalence rate 76.8% vs 66.6%; P<0.001). This association persisted after adjustment for confounding factors and was predominantly present in subjects with no to mild alcohol consumption. CONCLUSION: There is an independent association between hepatic steatosis and carotid atherosclerotic plaques. Metabolic changes due to nonalcoholic fatty liver disease may explain this relationship. PMID- 15793880 TI - Effect of interferon alpha2b plus ribavirin treatment on selected growth factors in respect to inflammation and fibrosis in chronic hepatitis C. AB - AIM: Growth factors (GF) that participate in regeneration and apoptosis have an important role in chronic liver diseases. We analyzed serum GF concentration during antiviral treatment and correlated it with morphological liver failure in chronic hepatitis C. METHODS: The levels of GF were determined in sera by ELISA method in 0, 16, 32 and 48 wk of therapy in 40 patients treated with IFNalpha2b (9 MU sc/wk) and RBV (1.2 g/d) and in 25 healthy subjects. Blind liver biopsies were done before treatment with histological grading and staging examination. RESULTS: The hepatocyte growth factor (HGF) and epidermal growth factor (EGF) were markedly elevated prior the treatment and decreased during the therapy, although they did not reach the normal level. In non-responding (NR) patients, HGF and EGF were higher than that in responders (R), however differences were not significant. Before the treatment thrombopoietin (TPO) level was significantly lower in R than in NR (P<0.03). Platelet-derived growth factor (PDGF) concentration was lower in chronic hepatitis C than in healthy subjects and decreased during the treatment. A significant positive correlation was observed between inflammatory activity in the liver tissue and the concentration of HGF (in R: r = 0.4, in NR: r = 0.5), TPO (R: r = 0.6), and a significant negative correlation between this activity and EGF (R: r = -0.6) and PDGF (R: r = -0.5). Serum HGF concentration was higher in more advanced fibrosis (R: r = 0.5, P<0.05; NR: r = 0.4, P<0.03). CONCLUSION: The decrease in PDGF can be an effective prognostic marker of the treatment and HCV elimination. Decreasing HGF, EGF, and PDGF can influence the inhibition of inflammatory and fibrotic processes in the liver during the antiviral treatment. PMID- 15793881 TI - Trial of a novel endoscopic tattooing biopsy forceps on animal model. AB - AIM: To tattoo gastric mucosa with a novel medical device which could be used to monitor and follow-up gastric mucosal lesions. METHODS: Combining endoscopic biopsy with sclerotherapy injection, we designed a new device that could perform biopsy and injection simultaneously. We performed endoscopies on a pig by using a novel endoscope tattoo biopsy forceps for 15 mo. At the same time, we used two step method combining sclerotherapy injection needle with endoscopic biopsy. The acuity, inflammation and duration of endoscopy were compared between two methods. RESULTS: Compared with the old two-step method, although the inflammation induced by our new device was similar, the duration of procedure was markedly decreased and the acuity of tattooing was better than the old two-step method. All characteristics of the novel device complied with national safety guidelines. Follow-up gastroscopy after 15 mo showed the stained site with injection of 1:100 0.5 mL of India ink was still markedly visible with little inflammatory reaction. CONCLUSION: Endoscopic tattooing biopsy forceps can be widely used in monitoring precancerous lesions. Its safety and effectiveness has been established in animals. PMID- 15793882 TI - Genetic distinctions between autoimmune hepatitis in Italy and North America. AB - AIM: Our goals were to analyze the known genetic predispositions for autoimmune hepatitis (AIH) in AIH Italian population and to compare them with North American counterparts. METHODS: Human leukocyte antigens (HLA) B8, C7, DR3, DR4, DR7, DR11, DR13, DQ2 and the B8-DR3-DQ2 phenotype were determined by microlymphocytotoxicity and polymerase chain reaction in 74 Italian patients (57 with type 1 and 17 with type 2 AIH) and 149 North American patients with type 1 AIH, and in adequate controls. RESULTS: B8-DR3-DQ2 occurred more frequently in Italian patients with type 1 AIH than in Italian controls (30% vs 7%, P<0.0001), but less frequently than in North American counterparts (30% vs 48%, P = 0.02). DR4 occurred less frequently in Italian patients with type 1 AIH (23% vs 43%, P = 0.01) and in controls (16% vs 34%, P = 0.0003) than in North American counterparts. No differences were found in alleles' frequency between type 1 and type 2 Italian AIH patients. DR11 had a frequency lower in type 1 Italian AIH patients than controls (17% vs 35%, P = 0.01). CONCLUSION: HLA DR4 is not associated with AIH in Italy. The known HLA risk factors for AIH occur similarly in Italian patients with type 1 and type 2 AIH, and they are less frequent than in North American patients. B8-DR3-DQ2 is the predominant phenotype of type 1 AIH also in Italy, and HLA DR11 may be a regionally distinctive protective factor against type 1 AIH. PMID- 15793883 TI - Cytochrome p450 2E1 polymorphisms and the risk of gastric cardia cancer. AB - AIM: Genetic polymorphisms of drug-metabolizing enzymes have recently been shown to affect susceptibility to chemical carcinogenesis. Cytochrome P450 2E1 (CYP2E1) enzyme catalyzes the metabolism of many procarcinogens, such as N-nitrosamines and related compounds. The gene coding for this enzyme is polymorphic and thus may play a role in gastric cardia cancer (GCC) etiology. In this hospital-based case-control study, we evaluate the relationship between genetic polymorphisms of CYP2E1 and the risk of GCC. METHODS: The study subjects comprised 159 histologically confirmed GCC cases identified via hospital cancer registry and surgical records at five hospitals in Fuzhou, Fujian Province, China, between April and November 2001. Controls were 192 patients admitted to the same hospitals for nonmalignant conditions. The genotypes of CYP2E1 were detected by a PCR-based RFLP assay. The odds ratios were estimated by logistic regression analyses and were adjusted for potential confounding factors. RESULTS: The distribution of three genotypes of CYP2E1 in GCC cases and controls was significantly different (chi(2) = 16.04, P<0.01). The frequency of the CYP2E1 (c1/c1) genotype in GCC cases and controls was 60.4% and 40.1%, respectively. The CYP2E1 (c1/c1) genotype was associated with an increased risk for GCC (the adjusted (OR) was 2.37, 95% confidence interval (CI): 1.52-3.70). Subjects who carried the CYP2E1 (c1/c1) genotype and were habitual smokers were at a significantly higher risk of developing GCC (OR = 4.68, 95%CI: 2.19-10.04) compared with those who had the CYP2E1 (c1/c2 or c2/c2) genotype and did not smoke. CONCLUSION: These results suggest that the CYP2E1 genotype may influence individual susceptibility to development of GCC, and that the risk increases significantly in smokers. PMID- 15793884 TI - Endoscopic ablation of Barrett's esophagus using high power setting argon plasma coagulation: a prospective study. AB - AIM: This prospective study evaluated the effectiveness of 90 W argon plasma coagulation (APC) for the ablation of Barrett's esophagus (BE) that is considered to be the main risk factor for the development of esophageal adenocarcinoma. METHODS: The results from 25 patients, observed at the First Department of General Surgery, University of Verona, Italy, from October 2000 to October 2003, who underwent APC for histologically proven BE were prospectively analyzed. RESULTS: The ablation treatment was completed in all the patients but one (96%). The mean number of APC sessions needed to complete ablation was 1.6 (total number: 40). The eradication was obtained in the majority of cases by one session only (60%), two sessions were required in 24% of the cases and three or more in 16%. About 43% of the sessions were complicated. Retrosternal pain (22.5%) and fever (17.5%) were the most frequent symptoms. Only one major complication occurred, it was an hemorrhage due to ulcer formation on the treated esophagus that required urgent endoscopic sclerosis and admission. The follow-up was accomplished in all the patients with a mean period of 26.3 mo and 20 patients (84%) with a follow-up period longer than 24 mo. Only one patient showed a relapse of metaplastic mucosa 12 mo after the completion of ablation. The patient was hence re-treated and now is free from recurrence 33 mo later. CONCLUSION: High power setting (90 W) APC showed to be safe and effective. The effects persist at a mean follow-up period of two years with a comparable cost in term of complications with respect to standard power settings. Further studies with greater number of patients are required to confirm these results and to assess if ablation reduces the incidence of malignant progression. PMID- 15793885 TI - Effect of omeprazole on symptoms and ultrastructural esophageal damage in acid bile reflux. AB - AIM: To value whether omeprazole could induce the healing of DIS and regression of symptoms in patients with DGER. METHODS: We enrolled 15 symptomatic patients with a pathological esophageal 24-h pH-metry and bilimetry. Patients underwent endoscopy and biopsies were taken from the distal esophagus. Specimens were analyzed at histology and transmission electron microscopy (TEM). Patients were treated with omeprazole 40 mg/d for 3 mo and then endoscopy with biopsies was repeated. Patients with persistent heartburn and/or with an incomplete recovery of DIS were treated for 3 more months and endoscopy with biopsies was performed. RESULTS: Nine patients had a non-erosive reflux disease at endoscopy (NERD) while 6 had erosive esophagitis (ERD). At histology, of the 6 patients with erosive esophagitis, 5 had mild esophagitis and 1 moderate esophagitis. No patients with NERD showed histological signs of esophagitis. After 3 mo of therapy, 13/15 patients (86.7%, P<0.01) showed a complete recovery of DIS and disappearance of heartburn. Of the 2 patients treated for 3 more months, complete recovery of DIS and heartburn were achieved in one. CONCLUSION: Three or 6 mo of omeprazole therapy led to a complete regression of the ultrastructural esophageal damage in 86.7% and in 93% of patients with DGER, NERD and ERD respectively. The ultrastructural recovery of the epithelium was accompanied by regression of heartburn in all cases. PMID- 15793886 TI - Adenocarcinoma and infection in a solitary hepatic cyst: a case report. AB - Solitary non-parasitic liver cysts are being increasingly diagnosed due to the increased use of abdominal sonography. The majority of solitary liver cysts are asymptomatic; however, there are some complications which include infection, perforation, spontaneous hemorrhage, obstructive jaundice and neoplastic degeneration. In some cases a cystic liver lesion may mimic a tumor and is difficult to differentiate with standard imaging studies or fine needle aspiration cytology. Here in, we report a case of adenocarcinoma arising in a solitary hepatic cyst complicated with Klebsiella pneumoniae infection. High levels of CEA in the cyst fluid levels suggested malignancy, which was confirmed by pathology of the resected specimen. PMID- 15793887 TI - Small-bowel perforation caused by fish bone. AB - A diagnosis of small-bowel perforation, caused by a sharp or pointed foreign body, is rarely made preoperatively because the clinical symptoms are usually nonspecific and can mimic other surgical conditions, such as appendicitis and diverticulitis. We report the case of a 62-year-old woman who experienced severe pain in the right iliac fossa and fever for about five days before arrival at our hospital. The presumptive diagnosis was acute purulent appendicitis and an emergency appendectomy was planned. Swelling and erythema were noted in a segment of the small bowel in the lower right abdomen. A tiny pointed object was found penetrating the inflamed portion of the bowel, which proved to be a sharp fish bone (gray snapper). The bone was removed, followed by segmental resection of the terminal ileum and ascending colon. The postoperative course was uneventful. PMID- 15793889 TI - HIV: more voluntary testing recommended. AB - Two research articles and an editorial in the New England Journal of Medicine recommend routine HIV testing for most of the U.S. population. The goal is to start treatment early when it can be more effective--and also to reduce transmission from the hundreds of thousands of Americans who do not know they have HIV. PMID- 15793888 TI - Isolated varices over hepatic flexure colon indicating superior mesenteric venous thrombosis caused by uncinate pancreatic head cancer--a case report. AB - Very rare cases of varices involving right side colon were reported. Most of them were due to cirrhotic portal hypertension or other primary causes. No report case contributed to pancreatic cancer. Here, we reported a case of uncinate pancreatic cancer with the initial finding of isolated hepatic flexure colon varices. Following studies confirmed isolated varices involving hepatic flexure colon due to pancreatic cancer with occlusion of superior mesenteric vein. From this report, superior mesenteric vein occlusion caused by uncinate pancreatic head cancer should be considered as a differential diagnosis of colon varices. PMID- 15793890 TI - Massachusetts project cuts a wide swath of care. Nontraditional testing sites is main focus. AB - Massachusetts organization provides rapid HIV testing services to clients at homeless shelters, domestic violence shelters, youth groups, churches, a community college, and other sites as part of a CDC-funded demonstration project. PMID- 15793891 TI - [Evidence based pneumology: 3rd update workshop to the SPLF. Management of spontaneous and primary pneumothorax]. PMID- 15793892 TI - [Evidence based pneumology: 3rd update workshop to the SPLF. Diagnosis of pleurisy (except cancer)]. PMID- 15793893 TI - [Evidence based pneumology: 3rd update workshop to the SPLF. Smoking: from prevention to weaning]. PMID- 15793894 TI - [Legionellosis: definition, diagnosis, treatment]. PMID- 15793896 TI - Bibliography. Current world literature. Pathophysiology of hypertension. PMID- 15793895 TI - [Clinic assistence for administrative and organizational support of hospital physicians]. PMID- 15793897 TI - [Use of zoledronic acid for high risk prostate cancer patients]. PMID- 15793898 TI - Bibliography. Current world literature. Sexually transmitted diseases and urinary tract infections. PMID- 15793899 TI - Abuse of prisoners at Abu Ghraib. PMID- 15793900 TI - Abuse of prisoners at Abu Ghraib. PMID- 15793901 TI - Abuse of prisoners at Abu Ghraib. PMID- 15793902 TI - [Individuality of emotional stress]. AB - The author postulates a need of individual evaluation of experimental data characterizing emotional stress. In typical conflict situations inducing emotional stress, there are animals resistant or predisposed to disturbance of different physiological functions. Prognostic criteria of individual resistance of animals to stressors are presented. Stress resistance depends in large on neuromediators and neuropeptides content, in particular Substance P, a peptide responsible for delta-sleep, and beta-endorphin, in brain structures and peripheral tissues. It is emphasized that individual estimation of clinical indices of emotional stress is needed. PMID- 15793903 TI - [Moderate cognitive impairment syndrome in cerebral vascular insufficiency]. AB - Frequency of clinical and psychological peculiarities of moderate (mild) cognitive impairment (MCI) in cerebral vascular insufficiency (CVI) was studied in 42 patients, mean age 67.7 +/- 7.7 years. Along with neurological examination, a battery of psychiatric and neuropsychological scales and tests was used. MCI was diagnosed in 28 (66.7%) patients. In 20 patients (47.6%), cognitive hypoactivity was most prominent related to impaired neurodynamics and regulation of voluntary activity which may reflect subcortical and frontal lobes functions. In 8 patients (20.1%), there was a prominent memory impairment characteristic of hippocampus lesion. The authors suggest the presence of concomitant neurodegeneration in patients with amnesic MCI. PMID- 15793904 TI - [Use of cognitive evoked potentials P300 in diagnosis of affection of the higher mental functions after surgery on the heart in conditions of cardiopulmonary bypass]. AB - Parameters of cognitive evoked potentials P300 were studied in 40 patients with ischemic heart disease subjected to coronary artery bypass graft surgery in conditions of cardiopulmonary bypass before and 7-9 days after operation. Patients underwent also neurological and neuropsychological examination. The most significant P300 parameters proved to be P3 and N2 latency, an increase of which correlated with the level of developing cognitive deficiency. A neuroprotective effect of Tracilol during the surgery was confirmed. In patients receiving this drug, cognitive deficiency and changes of P300 parameters were less pronounced. A study of cognitive evoked potentials P300 may be recommended for detecting early cognitive dysfunction and estimation of neuroprotective therapy in patients operated on in conditions of cardiopulmonary bypass. PMID- 15793905 TI - Cholera vaccine might work in AIDS patients. PMID- 15793906 TI - Vaccine. Official: tight budget could affect AIDS research. PMID- 15793907 TI - Court upholds patient's charge for HIV testing. PMID- 15793908 TI - Activism. Students rally at Capitol. PMID- 15793909 TI - Activism. Tech elite rallied to fight global AIDS and poverty. PMID- 15793910 TI - HIV testing order reversed for man who wrote explicit letters. PMID- 15793911 TI - Arbitration is ordered in HIV-drug patent case. PMID- 15793912 TI - Discrimination. State Department's anti-HIV hiring policy under scrutiny. PMID- 15793913 TI - Discrimination. Lambda Legal campaign urges antidiscrimination at work. PMID- 15793914 TI - [Acute oesophageal necrosis]. AB - The case of acute oesophageal necrosis in 56-year-old patient with tumor dissemination is presented. Acute endoscopy was performed after several hours lasting haematemesis and melena. Endoscopy revealed acute oesophageal necrosis in more than 2/3 of the oesophagus. Patient was treated with proton pump blockers and the total parenteral sustenance was introduced. Three days after the hospital admission the patient died in the septical metabolic disorder resulting from a disseminated spinocellular carcinoma of cervix uteri. Autopsy confirmed necrosis of oesophageal mucosa penetrating into the muscularis mucosae. Acute oesophageal necrosis is a rare disease, which is characteristic by the endoscopic finding of "black oesophagus", histological image of mucous membrane necrosis and by unknown aetiology. The article gives an overview of contemporary knowledge on the clinical, endoscopic and histologic images. The finding of "black oesophagus", which has been reported by several endoscopic units, has highly heterogeneous origin and it represents 0.0125 % (Moreto) till 0.28 % (Augusto) of cases. Our department has recorded 2 cases from 24271 of endoscopies performed in the last 8 years. PMID- 15793915 TI - Oral pathology quiz # 43. Case number 1. Fibrous dysplasia. PMID- 15793916 TI - Legal status of tooth whitening products. PMID- 15793917 TI - Fact sheet on Georgia's physician training programs. PMID- 15793918 TI - 2004 survey of physicians 50 to 65 years old. PMID- 15793919 TI - Physician supply and demand indicators in Georgia: 2003 GME exit survey. PMID- 15793920 TI - The effect of rising medical liability premiums on physician supply and access to medical care in Georgia. PMID- 15793921 TI - Information from your family doctor. Vertigo--a type of dizziness. PMID- 15793922 TI - Information from your family doctor. What should I know about cholesterol? PMID- 15793924 TI - 76 things you should know about the new Medicare drug discount cards. PMID- 15793925 TI - Into or beyond concept analysis in holistic nursing? PMID- 15793927 TI - Retraction. PMID- 15793926 TI - Discourse on concept analysis. PMID- 15793928 TI - Introduction and ISS assembly description. PMID- 15793929 TI - ISS crew evolution. PMID- 15793930 TI - A tour of the ISS. PMID- 15793931 TI - Over the tiniest patients, big ethical questions: fetal surgery's growing reach raises issues of need and risks. PMID- 15793932 TI - How to use the ISS. PMID- 15793933 TI - Persistent anemia in a renal transplant recipient. PMID- 15793935 TI - Space exploration and applications in the year 2025. PMID- 15793934 TI - Life science research in space brings health on Earth. PMID- 15793936 TI - The RESIST trials--superiority of tipranavir over other PIs. PMID- 15793937 TI - Perceived adverse effects of antiretroviral therapy. AB - Adverse effects from antiretroviral therapy (ARV) for HIV are associated with medication nonadherence. The purposes of this study were to explore group differences in the reporting of adverse effects, identify individual adverse effects that are linked to nonadherence, and to explore the role of coping in the relationship between adverse effects and adherence. Cross-sectional interviews of 2,765 HIV-positive adults on ARV therapies in four U.S. cities were performed using a computerized assessment of self-reported adverse effects, coping self efficacy, and adherence. There were no gender differences in the rate or severity of adverse effects reported. Latino respondents reported more adverse effects than either White or African Americans. Those taking a protease inhibitor (PI) reported a higher rate and greater severity of adverse effects. Older participants reported fewer adverse effects despite being more likely to be on a regimen containing a PI. Respondents with less than 90% adherence reported greater numbers and severity of adverse effects overall. In multivariate analyses, nausea, skin problems, vomiting, and memory adverse effects were independently related to less than 90% adherence over the prior three days. Coping moderated the relationship between nausea and adherence such that individuals who reported lower coping self-efficacy and experienced nausea were at increased risk for nonadherence, regardless of the length of time on the current ARV regimen. Women and men are similar in their overall reports of adverse effects, and Latinos report more adverse effects to ARVs than White or African American patients. Specific adverse effects (skin problems, memory problems, vomiting, and nausea) are more likely than others to be associated with missing ARV medications. Increasing adaptive coping self-efficacy among patients experiencing nausea may be a particularly effective strategy in increasing medication adherence. PMID- 15793938 TI - Use of implantable cardioverter-defibrillators for prevention of sudden death in patients at high risk for ventricular arrhythmia. PMID- 15793939 TI - Physician responsibility. PMID- 15793940 TI - Proper credit to surgical pioneers. PMID- 15793942 TI - Extracorporeal shock wave treatment for chronic plantar fasciitis. PMID- 15793941 TI - Evaluation of the safety and immunogenicity of pneumococcal seven-valent conjugate vaccine (Prevenar) administered in previously unvaccinated Spanish children aged 24 to 36 months. AB - This study evaluates the safety and immunogenicity of pneumococcal seven-valent conjugate vaccine (Prevenar) in 115 children, aged 2-3 years (24-36 months), who have not been previously vaccinated with Prevenar. RESULTS: SAFETY: As for local reactions, 40% of children reported erythema, 32.2% induration and 39.1% tenderness at the injection site. Regarding systemic reactions, fever > or 38 C was recorded in 7% of patients. Other commonly reported events were decreased appetite (24.3%), restlessness (20%), and fussiness (18.3%). IMMUNOGENICITY: After vaccination, more than 98% of the subjects achieved antibody levels of > or = 0.15 microg/mL for all seven serotypes and more than 95% achieved antibody levels > or = 0.50 microg/mL for all serotypes. CONCLUSIONS: Pneumococcal seven valent conjugate vaccine (Prevenar) was safe, well tolerated and highly immunogenic when administered in previously unvaccinated children aged 14-36 months. PMID- 15793944 TI - NSF launched for long-term neurological conditions. PMID- 15793943 TI - Study of methanogenesis during bioutilization of plant residuals. AB - The waste management strategy for the future should meet the benefits of human safety, respect principles of planet ecology, and compatibility with other habitability systems. For these purposes waste management technologies relevant to application of the biodegradation properties of bacteria are of great value. Biological treatment method is based on the biodegradation of organic substances by various microorganisms.The objectives of our study were: to evaluate the effectiveness of microbial biodegradation of vegetable non-edible residual, using artificial inoculum, and to study the peculiarities of biogas, and possibilities of optimizing or reducing the share of methane. The diminution rate of organic gained 76% from initial mass within 9 days of fermentation. The biogas production achieved 46 l/kg of substrate. The microbial studies of biodegradation process revealed the following peculiarities: (i) gradual quantitative increase of Lactobacillus sp. (from 10(3) to 10(5) colony-forming units (CFU) per ml); (ii) activation of Clostridia sp. (from 10(2) to 10(4) CFU/ml); and (iii) elimination of aerobic conventional pathogens (Enterobacteriaceae, Protea sp., Staphylococci). Chromatography analysis revealed the constant presence of carbon dioxide (up to 90.9%). The methane content measures revealed traces 0.1-0.4%. However, when we optimized the methane production in "boiling layer" using methanogenic granules, the amount of methane in biogas reached 80-90%. Based on the results obtained the artificial inoculum was created which was capable of initiating biodegradation of vegetable wastes. This inoculum consisted of active sludge adapted to wastes mixed with excretea of insects which consume plant wastes. Using this inoculum the biodegradation process takes less time than that using active sludge. Regulation of methane concentration from traces to 90% may be achieved by adding methane reactor to the plant digester. PMID- 15793945 TI - Will agenda for change leave nurses short changed? PMID- 15793946 TI - A voice from primary care. PMID- 15793947 TI - The visitors. PMID- 15793948 TI - Hearing aid. PMID- 15793949 TI - Learning to breathe again. PMID- 15793951 TI - Lichen planus. PMID- 15793950 TI - Ovarian cancer: addressing a continuing health care challenge. AB - Ovarian cancer is difficult to diagnose and to manage. Early detection and successful treatment continue to challenge health care professionals, although over the past 20 years there have been modest improvements in overall outcomes. The charity WellBeing of Women is running a campaign called 'Breaking the Silence' to focus on the issue of ovarian cancer. PMID- 15793952 TI - Silver sulphadiazine. PMID- 15793953 TI - An evaluation of tea tree oil as an alternative microbicide. AB - Tea tree oil is a popular ingredient in complementary medicines and beauty products. This literature review reveals some evidence of efficacy as an antiseptic, antibacterial and antifungal in topical application but also highlights dermatitis as a potential side-effect. PMID- 15793954 TI - Best practice for managing patients' postoperative pain. AB - This article describes the development of a best practice statement on postoperative pain management. It examines why this area was considered a priority, the development process, the content of the statement and the recommendations emerging from it. PMID- 15793955 TI - Appreciating the importance of oral health in all areas of nursing. PMID- 15793956 TI - Identifying the characteristics of children with pressure ulcers. AB - AIM: To identify the characteristics of children with pressure ulceration so that those at risk may be identified in future. METHOD: A multicentre survey was undertaken in 11 hospitals to ascertain the incidence, severity and position of pressure ulcers in paediatric inpatients. RESULTS: The most frequently reported ulcer grade involved blistering of the skin or a superficial skin break. The most frequent site was the sacrum or buttocks, heel, thigh, ear, occipital scalp, malleolus and spine. In 50 per cent (n=27) of children, pressure ulcers could be associated with equipment pressing or rubbing on the skin. CONCLUSION: Risk assessment tools designed specifically for children are needed, and more research should be undertaken to confirm the characteristics that increase children's risk of pressure ulceration and to establish the significance of individual characteristics. PMID- 15793957 TI - Using your time more effectively. PMID- 15793958 TI - Changes create new nursing careers. PMID- 15793959 TI - The modern world loves the idea of empowerment. PMID- 15793960 TI - Backing the ban. The Female Genital Mutilation Act makes it illegal to take girls abroad to be mutilated. PMID- 15793962 TI - Grand designs. Nurses have a reputation for being innovative. PMID- 15793961 TI - Smart cookies. Access to information technology has been a contentious issue for nurses, but there is enthusiasm over its benefits for patients. PMID- 15793963 TI - Mothers' milk. Give premature babies the best nutritional start in life. PMID- 15793964 TI - Treatment by the book. Self-help books are increasingly being prescribed by GPs instead of medication. PMID- 15793965 TI - Face the music. Avoiding difficult situations with colleagues will only make matters worse. PMID- 15793966 TI - Playing for keeps. Improvement in nurse recruitment must be matched by retention. PMID- 15793967 TI - Psychological assessment during cardiac rehabilitation. AB - BACKGROUND: Depression and anxiety are common after myocardial infarction (MI) and cardiac surgery and may adversely affect outcome. A comprehensive cardiac rehabilitation service needs to include psychological input as well as education and exercise training. To address this issue the Bromley Hospitals NHS Trust (BHT) cardiac rehabilitation nursing team introduced the Hospital Anxiety and Depression Scale (HADS) into its cardiac rehabilitation service. Over a 21-week period, 130 patients attending a cardiac rehabilitation nursing assessment completed a HADS. CONCLUSION: Audit of the HADS scores revealed that post-MI patients waiting longer than six months for cardiac rehabilitation had the highest levels of 'significant' anxiety (HADS score >10). Results suggest that timely entry into a cardiac rehabilitation programme helps to reduce the HADS scores of post-MI patients. The HADS was shown to be an effective tool for assessing the psychological wellbeing of patients attending cardiac rehabilitation. It enables those with significant anxiety and/or depression to be identified and offered further assessment by a cognitive behavioural therapist. PMID- 15793968 TI - Cardiovascular care of patients with Marfan syndrome. AB - Marfan syndrome is a genetic disorder with potentially fatal cardiovascular complications. These risks can be reduced by early diagnosis, appropriate review and timely intervention. Care of patients with Marfan syndrome should be multidisciplinary and multiprofessional. This article reviews the cardiovascular aspects of care and the nursing role. PMID- 15793969 TI - Practical guidance on sexual lifestyle and risk. AB - This article advocates using a critical thinking framework as a health promotion tool to help nurses promote thought and discussion in relation to sexual lifestyles. The aim is to encourage clients or confidants to assess sexual risk for themselves and others, and to promote safe sex. PMID- 15793970 TI - Prescription drugs: uses and effects. PMID- 15793971 TI - Starring role. Preparing for job interviews is often an ad hoc affair for nursing students. PMID- 15793972 TI - How technology offers alternative methods of qualitative analysis. PMID- 15793973 TI - Qualitative data analysis using data displays. AB - The amount of data generated in qualitative research can be difficult to manage. In this paper Tracey Williamson and Andrew Long discuss how the use of data displays can improve data management and also how the process can help to make the routes from raw data to research findings in qualitative research more transparent. Data displays can take several forms but share the benefit of helping to condense large amounts of data into more manageable forms. They can also help to convey information in a visually stimulating format where presentation time or column space may be limited. PMID- 15793974 TI - Analysing audio-recorded data: using computer software applications. AB - Conventional methods of qualitative data analysis require transcription of audio recorded data prior to conduct of the coding and analysis process. In this paper Alison Hutchinson describes and illustrates an innovative method of data analysis that comprises the use of audio-editing software to save selected audio bytes from digital audio recordings of meetings. The use of a database to code and manage the linked audio files and generate detailed and summary reports, including reporting of code frequencies according to participant code and/or meeting, is also highlighted. The advantage of using this approach in the analysis of audio-recorded data is that the process may be undertaken in the medium in which the data were collected. Though time-consuming, this process negates the need for expensive and time intensive transcription of recorded data. PMID- 15793975 TI - Telephone interviewing in healthcare research: a summary of the evidence. AB - Increasing numbers of healthcare researchers in the UK are using telephone interviews in their research. In this paper, Elizabeth Smith summarises the context in which the method is used to target purposive samples of service users or patient, carers, clinicians and other staff, or key individuals in the field of inquiry. The aim is to gain detailed and specific data about perceptions, experiences and views of healthcare services. It is useful to identify the circumstances under which the advantages and disadvantages of telephone interviewing have been realised to contextualise concerns about the quality of the data compared to other methods and provide practical advice for researchers considering using the method. PMID- 15793976 TI - Researching students: an ethical dilemma. AB - Students may be perceived as an accessible source for research, but their use as research subjects is not without ethical problems. These difficulties are increased when the researchers are also responsible for teaching the students they are researching. In this paper, Eileen Clark and Terence McCann reflect on the ethics of conducting research on their own students. They identify sources of ethical concern and discuss how these may be managed. Their experience is that lecturers can carry out research on their own students, even at small campuses, in an ethical manner They highlight the importance of opening a dialogue with the ethics committee at the institution. PMID- 15793977 TI - Preparing for the field: developing competence as an ethnographic field worker. AB - If ethnography can be considered as a form of work, then the rules and competencies of this work need to be learnt along with clear markers of progress to enable the ethnographer to gauge their developing competence. In this paper Fiona Murphy draws on the work of Benner (1984), which provides a framework for the proposition that there is a progression in developing competence as an ethnographic fieldworker from 'novice to expert'. This has implications for the preparation and support of novice fieldworkers in that these 'rules' and competencies should be made more explicit and transparent PMID- 15793978 TI - Historical research: examining documentary sources. AB - In this paper John Sweeney outlines key principles for conducting a study employing an historical design based on documentary analysis. It derives in part from his PhD study, in which he employs an historical design based on documentary analysis to investigate the delay in introduction of intellectual disability nursing into the Republic of Ireland from 1921 until 1958. This study examines the social, political and professional context to the decision to forgo the recognition of a professional nursing qualification and education process in intellectual disability until 1958. PMID- 15793979 TI - Blurred boundaries damage inter-professional working. AB - Everybody pays lip-service to the idea that healthcare professionals should collaborate in delivering care to patients. In this article, Rosemary Rushmer explores how a new methodology, connate theory could lead to improvements in our understanding of what makes healthcare teams tick. PMID- 15793980 TI - An eye for detail, a thirst for understanding and desire to improve the lives of people using health services. AB - Patrick Callaghan, Head of Department of Mental Health and Learning Disability, City University explains how a natural curiosity a stint in the banking sector and meeting an inspiring mentor were some stepping stones to a career in research PMID- 15793981 TI - Recognising harm. PMID- 15793982 TI - Co-ordination of complex care. PMID- 15793983 TI - The benefits of baby massage. PMID- 15793984 TI - Telemedicine: helping neurologically-impaired children to stay at home. PMID- 15793985 TI - Cystic fibrosis-related diabetes: practice challenges. PMID- 15793986 TI - Preparation for transition to adult diabetes services. PMID- 15793987 TI - Community children's nursing services in the 21st century. PMID- 15793988 TI - Hypercholesterolaemia: management and nursing implications. PMID- 15793989 TI - Emergency baptism by health professionals. PMID- 15793990 TI - Preparing for the worst. PMID- 15793991 TI - Relieving the pressure. PMID- 15793992 TI - Nursing a nation back heath. PMID- 15793993 TI - Picking up the pieces. PMID- 15793994 TI - Self-harm: reviewing psychological assessment in emergency departments. PMID- 15793995 TI - Spinal cord injury: acute care management. PMID- 15793996 TI - Acute experience. PMID- 15793997 TI - Periodical blues: journals and the hospital library. PMID- 15793998 TI - The second law of Ranganathan. PMID- 15793999 TI - JCAHO's Twelfth Annual Liaison Network Forum. PMID- 15794000 TI - Blog? PMID- 15794001 TI - Patient parent education and hospital librarians. PMID- 15794002 TI - What's up with nursing research? Nurses' perception of nursing research in a community hospital. PMID- 15794003 TI - Harnessing & celebrating the publishing energy of your organization. PMID- 15794004 TI - Supporting JCAHO accreditation activities. PMID- 15794005 TI - The era of the Americans With Disabilities Act. PMID- 15794006 TI - One (other) way to get started with electronic document delivery. PMID- 15794007 TI - NIN@PC: Nursing Information Needs at the Point of Care. PMID- 15794008 TI - The creative mind. PMID- 15794009 TI - "Information Rx" project launched in Virginia. AB - The Internet has introduced a number of challenges for physicians in their daily interactions with patients. Of these, one of the most profound challenges is the availability of unreliable sources of health information on the Internet. Ask any practicing physician to explain why this availability often hinders rather than aids patient education. First, the physician spends precious examination time explaining or proving to a patient why a resource found on the Internet is not reliable. The information may be inaccurate or out-of-date. And, second, some physicians are not aware of the best available Internet resource to which they can refer patients. PMID- 15794010 TI - New (York) renovations. AB - For busy solo medical librarians working in very busy hospitals, the job of maintaining top quality services while construction goes on over your head and under your feet is no small task! In the Mid-Hudson Valley area of New York State, several of our librarians have accomplished just such feats of endurance and the results have been well worth the effort. PMID- 15794011 TI - The adventures of mentoring an AHIP newbie. PMID- 15794012 TI - Presenting Loansome Doc. As part of a category-1 CME Improvement Program. PMID- 15794013 TI - Fit to drive. PMID- 15794014 TI - Internet interactions: adolescent health and cyberspace. PMID- 15794015 TI - Concerns about Family Medicine Forum. PMID- 15794016 TI - Training and rural physicians. PMID- 15794017 TI - Appreciating our health care system. PMID- 15794018 TI - Marijuana use and breastfeeding. AB - QUESTION: One of my breastfeeding patients is using marijuana to combat chronic pain. Is it safe for her to breastfeed? ANSWER: Lactating mothers should refrain from consuming cannabinoids. Advising mothers to discontinue breastfeeding if they cannot stop using cannabinoids must incorporate the known risks of formula feeding. Cannabinoid exposure through milk has not been shown to increase neonatal risk, but there are no appropriate studies of this. In every case, nursing babies should be closely monitored. PMID- 15794019 TI - Can you identify this condition? Melasma. PMID- 15794020 TI - Hyperkalemia associated with spironolactone therapy. PMID- 15794021 TI - Which older patients are competent to drive? Approaches to office-based assessment. AB - OBJECTIVE: To review three proposed approaches to office-based assessment of older drivers and to evaluate recommendations made about dementia and driving. QUALITY OF EVIDENCE: The American Medical Association's (AMA's) Physcian's Guide to Assessing and Counseling Older Drivers gives recommendations for office-based assessment of older patients' medical fitness to drive. Other approaches examined were those outlined in the sixth edition of Determining Medical Fitness to Drive produced by the Canadian Medical Association (CMA) and SAFE DRIVE. Recommendations for dementia and driving from these documents and other sources were reviewed. All evidence was level III. MAIN MESSAGE: The AMA document usefully identified ways to detect drivers at risk and key areas for assessment (vision, cognition, motor function). Recommendations on evaluating these areas require validation. .he CMA guide and SAFE DRIVE were overly broad in their recommendations. How best to detect cognitive impairment that tocld affect driving remains unclear. CONCLUSION: Office-based approaches to identifying older drivers who are either unsafe to drive or require more extensive evaluation need to be validated. PMID- 15794023 TI - Primary health care transition fund update. PMID- 15794022 TI - In-office evaluation of medical fitness to drive: practical approaches for assessing older people. AB - OBJECTIVE: To provide background for physicians'in-office assessment of medical fitness to drive, including legal risks and responsibilities. To review opinion based approaches and current attempts to promote evidence-based strategies for this assessment. QUALITY OF EVIDENCE: MEDLINE, EMBASE, CINAHL, PsyclNFO, Ageline, and Sociofile were searched from 1966 on for articles on health-related and medical aspects of fitness to drive. More than 1500 papers were reviewed to find practical approaches to, or guidelines for, assessing medical fitness to drive in primary care. Only level III evidence was found. No evidence-based approaches were found. MAIN MESSAGE: Three practical methods of assessment are discussed: the American Medical Association guidelines, SAFE DRIVE, and CanDRIVE. CONCLUSION: There is no evidence-based information to help physicians make decisions regarding medical fitness to drive. Current approaches are primarily opinion-based and are of unknown predictive value. Research initiatives, such as the CanDRIVE program of the Canadian Institutes of Health Research, can provide empiric data that would allow us to move from opinion to evidence. PMID- 15794024 TI - Centre for Studies in Primary Care: offering a laboratory of primary care practices. PMID- 15794025 TI - Teen angst. PMID- 15794026 TI - Effective stimulation of daily LH secretion by the combined treatment with melatonin and naloxone in luteal-phase ewes. AB - This study was designed to test the hypothesis that melatonin would intensify daily LH release after central blockade of the opiate receptors in sexually active ewes. The intracerebroventricular infusions of vehicle (control), melatonin, naloxone and melatonin in combination with naloxone were made in ewes in the luteal phase of the estrous cycle, from 2:00 P.M. to 5:00 P.M. Blood samples were collected from 11:00 A.M. to 8:00 P.M. at 10-min intervals. The mean plasma LH concentrations were measured before, during and after the infusions. The frequency and amplitude of LH pulses were determined during the whole experimental period. The LH concentrations recorded during melatonin or naloxone infusions were significantly higher than the concomitant concentration in vehicle infused animals. The mean LH pulse amplitude in melatonin- and naloxone-treated ewes was also significantly higher than in controls. The LH concentration measured during the combined infusion of melatonin and naloxone was significantly higher than that during vehicle infusion. The LH concentration recorded in turn after the treatment was significantly higher than the concomitant concentrations in vehicle-, melatonin- and naloxone-infused animals. The mean LH pulse amplitude in this group was significantly higher than in the vehicle-infused group. These results indicate that blockade of the opiate receptors within the CNS facilitated effective stimulation of daily LH secretion by exogenous melatonin. In conclusion, a relationship between melatonin and endogenous opioid peptides may be crucial in enabling melatonin to exhibit stimulatory action on LH secretion during the luteal phase of the estrous cycle in ewes. PMID- 15794027 TI - Spectral analysis of cerebellar activity after acute brain injury in anesthetized rats. AB - We investigated cerebellar electrocortical activity before and after unilateral brain injury in anesthetized rats. Spectral analysis of cerebellar activity was obtained by Fast Fourier Transformation. There was a dominance of delta frequency range, while the wide gamma range presented no more than 5% of the total mean power spectra of cerebellar activity before brain injury. A few minutes after brain injury and within the first 90 minutes, there was a decrease of total mean power spectra and a relative decrease of delta range power to about 30%, some increase of beta range, and an increase of gamma range to 20-25%. Relative increase of gamma range in the cerebellar mean power spectra was still present 120 min after the brain injury, while other changes started to diminish. We suggest that spectral changes within slow and fast (gamma) frequency ranges of cerebellar activity may be indicators of the brain state after acute injury. PMID- 15794028 TI - Increased prefrontal event-related current density after sleep deprivation. AB - To investigate how partial sleep loss affects temporal and spatial pattern of information flow, we analyzed sources of brain electrical activity during continuous attention test. Sixteen physicians recruited from the university hospitals participated in the study. Each participant served as his own control. All participants underwent two test sessions including the Stanford Sleepiness Scale (SSS), the Beck Depression Inventory (BDI), the Selective Reminding Test (SRT), and the Continuous Attention Test (CAT). The CAT items were used as stimuli in event-related potential (ERP) recordings. EEG was recorded from 21 electrodes, according to the international 10-20 system. The sources of bioelectrical activity were computed with low resolution electromagnetic tomography (LORETA). Estimated sleep time was significantly shorter on nights spent on duty than on nights of normal sleep at home. Sleep loss resulted in significant increase in SSS and BDI scoring, and impairment of immediate recall. Performance on the CAT remained relatively intact. Under the sleep loss condition compared to baseline, significant differences in brain activity occurred only for targets. Within the P1 time frame, sleep loss led to greater activation in the right Brodmann's area 9/10. For the N1 component, significant differences were localized on the lateral surface of the right frontal lobe, in Brodmann's areas 8 and 9. No significant effects of sleep deprivation on the P3 component were found. Our results are consistent with earlier data indicating that increased activation of the prefrontal cortex allows the maintainance of performance during periods of sleep loss. PMID- 15794029 TI - Behavioral responses of 129/Sv, C57BL/6J and DBA/2J mice to a non-predator aversive olfactory stimulus. AB - We examined the behavior of three inbred mouse strains (129/SvPasIco, C57BL/6J, and DBA/2J) exposed to an object soaked with the chemical component of the aversive scent (toluquinone odor) emitted by a myriapod species (Ommatoiulus sabulosus) in the presence of a predator. Subjects were exposed to the odor for three consecutive days. Behavioral responses to the toluquinone odor were characterized both by an approach phase of risk assessment and by a repeated series of approach-avoid episodes. Results indicate that toluquinone exposure reduced completely, and in a strain independent fashion, selected behaviors such as crouching, catching and eating object. Other responses were strain-dependent: the DBA (DBA/2J) strain displayed defensive burying at high levels, C57 (C57BL/6J) mice performed high levels of withdrawal while the 129/Sv (129/SvPasIco) strain showed also high levels of stretch attend posture. Compared to other tasks, this test is ethological, simple, cheap and is not affected by strain differences in appetitive-sensory responses, as shown by some strain independent responses. These features make this task as a good complement to any exploration-anxiety test battery. PMID- 15794030 TI - Ranking of memories and behavioral strategies in the radial maze. AB - New features of actual choice behavior and effortful information processing in rats were demonstrated in an eight-arm radial maze through modifications of a matching-to-sample task. Two attempts were allowed for a squad of hooded Sprague Dawley rats (n=7) for finding a reward in a testing phase of the task. The results showed flexibility and sooner learning to matching rule on the second testing attempt that was only later followed by an improvement of choice accuracy on the first attempt. "Hidden learning" on the second attempt could reflect memories and behavioral strategy, which were present, but not expressed on the first choice. The hypothesis was advanced that learning expressed on the second attempt reflects encoding of a matching rule, whereas improvement on the first choice reflects changes in the rank of acquired memories and behavioral strategy. A second experiment on the same squad of rats tested the ability of trained animals to rank already acquired memories. Following the introduction of the second sample to the study phase of the task, the rats learned to prefer to match the first sample in the testing phase, rearranging ranks of stored memories under the internal control of win-stay strategy. Alternative explanations of interference, trace decay and ranking were compared in order to account for the present results. PMID- 15794031 TI - Superimposition of illusory patterns with contrast variations. AB - In psychophysical experiments, a perceived length matching task was performed. The stimuli were made of two spatially superimposed illusory figures that differed in structure and luminance contrast but had the same length and coincided precisely, with their ends matched. The contrast of one of the figures was fixed, and that of the other varied. In experiments with stimuli viewed monocularly, the combined patterns produced illusions of perceived length, the strength of which varied with alterations of contrast of one of the figures. If the figures were presented separately to different eyes of the same subject, changes of contrast did not have a noticeable influence on the illusion's strength. When the two stimulus components were displaced spatially and shown side by side, the monoptic and dichoptic stimuli yielded different results as well. The illusion's strength increased with an increase of the distance between the figures when viewed monocularly, but remained invariable if the figures were presented separately to each eye. The results obtained in experiments with dichoptic stimuli suggest that stimulus length distortions may occur in the monocular retinocortical pathways. PMID- 15794032 TI - Visual and auditory attentional capture are both sluggish in children with developmental dyslexia. AB - Automatic multimodal spatial attention was studied in 12 dyslexic children (SRD), 18 chronological age matched (CA) and 9 reading level matched (RL) normally reading children by measuring reaction times (RTs) to lateralized visual and auditory stimuli in cued detection tasks. The results show a slower time course of focused multimodal attention (FMA) in SRD children than in both CA and RL controls. Specifically, no cueing effect (i.e., RTs difference between cued uncued) was found in SRD children at 100 ms cue-target delay, while it was present at 250 ms cue-target delay. In contrast, in both CA and RL controls, a cueing effect was found at the shorter cue-target delay but it disappeared at the longer cue-target delay, as predicted by theories of automatic capture of attention. Our results suggest that FMA may be crucial for learning to read, and we propose a possible causal explanation of how a FMA deficit leads to specific reading disability, suggesting that sluggish FMA in dyslexic children could be caused by a specific parietal dysfunction. PMID- 15794033 TI - Does electroconvulsive therapy (ECT) affect cognitive components of auditory evoked P300? AB - Electroconvulsive therapy (ECT), as a treatment tool for psychiatric disorders, is believed to be safe and effective. Nevertheless, it has a negative impact on cognitive functioning, especially on memory, causing both retrograde and anterograde amnesia. However, ECT effects on more subtle stages of information processing are not studied enough. Event-related potentials, and especially P300, are thought to reflect physiology of cognition. Thus, we aimed to evaluate the effects of ECT treatment on parameters of endogenous components (N2, P3) of the P300 potential. Seventeen patients suffering from schizophrenia, schizoaffective disorder and recurrent depressive disorder participated at the study. After the course of ECT, significant increase of N2 amplitude in parietal midline region and prolongation of P3 latency in frontal midline region, of which the magnitude positively correlated with the number of ECT procedures, have been obtained. PMID- 15794034 TI - Posterior parietal cortex and developmental dyslexia. AB - Dyslexia is defined as a specific reading disorder despite normal intelligence and conventional teaching. One of the most influential theories attempting to explain problems suffered by dyslexics assumes that dyslexia is caused by deficits of the magnocellular system. This system, generally responsible for processing fast sensory information, projects mostly to the parietal cortex. Consistent with this theory, dyslexics should have problems with tasks which specifically involve parietal cortex. In the article, we review data and show that, indeed, dyslexics have problems with fast attention shifts, show some symptoms of mild unilateral neglect syndrome and have abnormal saccadic and pursuit eye movements. Little is known about visuo-motor coordination and mental rotation, the tasks in which the parietal cortex is thought to play important roles. PMID- 15794036 TI - One world of humanity--and orthodontics. PMID- 15794035 TI - The structure and dynamics of normal language processing: insights from neuroimaging. AB - This paper reviews current psycholinguistic and neuroimaging evidence on language processing with particular focus on the relationship between production and comprehension. In the first part, different methods of psycholinguistic research are introduced and examples for psycholinguistic models (production: Levelt et al. 1999; comprehension: Friederici 2002) are sketched. In the second part, the neural correlates of semantic, phonological, and syntactic processing are reviewed. For semantics and phonology there seem to be different fronto-temporal networks which are shared in production and comprehension. The results for the processing of syntactic information are not entirely conclusive. Yet the data reveal that phonological strategies may be used in syntactic tasks. This finding opens the discussion of alternative, phonology-based strategies for language processing. Such strategies are accounted for by dual-route models featuring one direct and one indirect route which often involves phonological processing. This insight leads to some tentative conclusions about remediation strategies in dyslexics with selective (e.g., phonological) deficits. PMID- 15794037 TI - Arch development with trans-force lingual appliances. AB - Trans-Force lingual appliances are designed to correct arch form in patients with contracted dental arches. Interceptive treatment with this new series of pre activated lingual appliances offers new possibilities for arch development, in combination with fixed appliances. Palatal and lingual appliances insert in horizontal lingual sheaths in molar bands. No activation is required after the appliance is fitted, and this principle is extended to a series of appliances for sagittal and transverse arch development. Both sagittal and transverse appliances have additional components to achieve 3-way expansion where this is indicated. The invisible lingual appliances may be used in correction of all classes of malocclusion at any stage of development, from mixed dentition through permanent dentition, and this approach has wide indications in adult treatment. PMID- 15794038 TI - Bonding orthodontic brackets to porcelain using different adhesives/enamel conditioners: a comparative study. AB - AIM: To evaluate the use of new adhesive/primer materials, including an experimental self-etch primer and a cyanocrylate adhesive, to enhance the shear strength of orthodontic brackets bonded to porcelain surfaces. MATERIAL AND METHODS: Sixty porcelain maxillary central incisor teeth were used. The teeth were randomly divided into four groups: group 1, teeth were etched with 37% phosphoric acid and the brackets were bonded with a composite adhesive; group 2, teeth were microetched, hydrofluoric acid and silane applied, and then the brackets were bonded with a composite adhesive; group 3, an acid-etch primer was used, then the brackets were bonded with the same composite adhesive as in the first 2 groups; group 4, teeth were etched with 35% phosphoric acid and the brackets were bonded with the cyanoacrylate adhesive. RESULTS: The analysis of variance comparing the groups tested (F = 9.446) indicated that there was a significant difference between the 4 groups. The cyanoacrylate adhesive had the lowest shear bond strength (mean = 1.7 +/- 2.1 MPa), followed by the conventional bonding using a 37% phosphoric acid etch and composite (mean = 2.1 +/- 1.2 MPa). The use of Transbond after microetching, with the application of hydrofluoric acid and silane, provided the highest shear bond strength (mean = 5.5 +/- 2.7 MPa). Transbond used with the acid etch-primer had a lower bond strength (mean = 3.8 +/- 2.5 MPa), but was not significantly different from the microetch/hydrofluoric acid/silane group. CONCLUSION: The results indicated that the use of a phosphoric acid etch with either a cyanoacrylate or composite adhesive to bond orthodontic brackets to porcelain surfaces produced significantly lower shear bond strength. Self-etch primers produced higher but less consistent shear bond strength for bonding orthodontic brackets. The most reliable bonding procedure to porcelain surfaces is through microetching with the use of hydrofluoric acid and a silane coupler before bonding, but this also produces the greatest damage to the porcelain surface. PMID- 15794039 TI - Impacted mandibular second molar with extruded maxillary second molar: a case report. AB - AIM: To present a method for correction of an extruded maxillary second molar. METHODS: A female patient, 16 years of age, presented with an impacted mandibular second molar and an extruded opposing maxillary second molar. After placement of appliances in the maxilla, an elastic was hooked between the distal end of the extended lingual arch and the lingual button of the maxillary second molar. RESULTS: After sufficient intrusion of the maxillary second molar was achieved, the conventional method for traction of an impacted mandibular second molar was used. Neither root resorption nor undesirable periodontal problems were observed. CONCLUSION: The approach used in the present case is simple and efficient for both traction and retention of a maxillary molar. PMID- 15794040 TI - Critical issues concerning root resorption: a contemporary review. AB - Root resorption, an unwanted sequela of orthodontic treatment, has received much attention in the recent past. This is clearly evident from the enormous amount of literature published regarding this single iatrogenic issue. These research works should provide practitioners with great insight into the clinical, histologic, therapeutic, predictive, and genetic aspects as well as the molecular basis of the resorptive process. Yet the scientific community reading these reports is left with confounding questions requiring a proper organization of the published literature. This review is aimed at answering some critical issues concerning root resorption after orthodontic treatment. It also tries to update the theoretical information regarding the resorptive process, with an insight into recent human genetic and molecular-level research. PMID- 15794041 TI - Differential diagnosis of skeletal open bite based on sagittal components of the face. AB - AIMS: This study examined the morphology of skeletal open bite with respect to the sagittal components of the face. METHODS: The material consisted of cephalometric and hand-wrist films of 49 girls and 22 boys with skeletal open bite. The samples were grouped into Class I, II, and III facial types on the basis of ANB angle. In addition to conventional dentofacial variables, nasopharyngeal airway area was also measured on lateral headfilms. All measurements were examined by analysis of variance and Duncan test. Subsequently the factors leading to open bite were evaluated using multiple-regression analysis. RESULTS: Dentofacial morphology differed in the sagittal components of skeletal open bite, and the differences were most obvious between the Class II and Class III open bite groups. Posterior maxillary dentoalveolar height and mandibular incisor inclination were important factors in the development of open bite in the skeletal Class I and Class II open bite groups, while in the skeletal Class III open bite group, the nasopharyngeal airway and the gonial angle were involved. CONCLUSION: Sagittal components of skeletal open bite should be considered in the differential diagnosis and treatment planning of such cases. PMID- 15794042 TI - Craniofacial morphology of skeletal class III patients before treatment and growth-related changes during treatment with a maxillary protraction appliance: a comparison of orthodontic and surgical cases. AB - AIMS: To examine differences in the craniofacial morphology of skeletal Class III patients before treatment and growth-related changes during treatment with a maxillary protraction appliance (MPA) between patients who were treated only orthodontically (orthodontic group) and those who eventually received orthognathic surgery (surgical group). METHODS: The orthodontic and surgical groups consisted of 6 boys and 9 girls and 4 boys and 6 girls, respectively. Lateral cephalograms taken at the first visit and at the end of treatment with the MPA were measured. The Mann-Whitney U test and Wilcoxon signed-rank test were used. RESULTS: Before treatment, the Wits appraisal was significantly smaller and the gonial angle was significantly larger in the surgical group. During treatment, both the SNA and ANB angles were significantly increased in the orthodontic group, whereas only the SNB angle was significantly increased in the surgical group. Moreover, significant lingual inclination of the mandibular incisors was found in the orthodontic group. CONCLUSION: The Wits appraisal and gonial angle were useful for distinguishing between the two groups at the first visit. Facilitation of forward maxillary growth, modification of the direction of mandibular growth, and lingual tipping of the mandibular incisors may be important ways to avoid orthognathic surgery. PMID- 15794044 TI - Utilizing a directional force system in class II mechanics. AB - The concepts of esthetics, function, and health must be supported by sound diagnosis, treatment planning, treatment timing, and mechanotherapy for each orthodontic patient. The treatment of a patient with a Class II, Division 1-type malocclusion is shown in this case report. The treatment sequence of complete Class II mechanotherapy and a directional force system are emphasized. PMID- 15794043 TI - Bones, teeth, and genes: a genomic homage to Harry Sicher's "Axial Movement of Teeth". AB - AIM: The model of the unopposed rodent molar was used to study the morphologic and genetic mechanisms of tooth eruption. METHODS: Left maxillary molar teeth of 12-day-old Swiss-Webster mice were extracted under anesthesia, and mandibular molars were allowed to supererupt. To trace areas of tissue remodeling and to determine areas of new tissue formation, mice were injected with fluorescent dyes, tetracycline, alizarin red, and calcein blue. Subsequent to sacrifice, mandibular tissue blocks were prepared for ultrathin ground sections, fluorescent microscopy, and von Kossa's mineral detection procedure. A second set of specimens was prepared for RNA extraction and microarray analysis. RESULTS: The data established significant eruption of first and second mandibular mouse molars 12 days after complete extraction of antagonists, exceeding the control side by 0.13 mm. Labeled tissue sections revealed significant amounts of new bone and cementum apposition on the unopposed side compared to the control side, as revealed by fluorescent markers and ultrathin ground sections. Microarray transcript level comparisons between the experimental and the control groups demonstrated significant (more than twofold) increase in gene expression of elastin and tenascin C extracellular matrix proteins; brevican, lumican, and biglycan proteoglycans; as well as fibroblast growth factor 9. CONCLUSION: In this study, the authors have established the unopposed mouse molar as a model to study tissue dynamics during the axial movement of teeth. The data indicated significant new formation of bone and cementum in tandem with increased expression of extracellular matrix-related genes. PMID- 15794045 TI - Optimal mechanics for mandibular molar uprighting. PMID- 15794046 TI - Orthodontic eduation in the USA: the challenge and an answer [interview by Samir E. Bishara]. PMID- 15794047 TI - "Namaskar" in Nepali means "greetings with both hands folded". PMID- 15794048 TI - [Correction of presbyopia in pseudophakic patients under 60 years of age]. AB - Objective of the study was to show the mean values of correction for near vision and to discuss the presbyopic correction in pseudophakic patients. Setting was the Eye department where authors work. Inclusion criteria for 55 patients were native or corrected distant vision of 0.8-1.0 on Snellen's chart; 0,6 on Jagger's chart for near vision; round pupil and good position of the implant. Biometry of the anterior chamber depth with Alcon biophysics during distant and near vision was performed in our study. "Hi square" test was carried out and it was concluded that patients younger than 59 years (41 eyes) had median correction of +2.0 dsph, while patients older than 60 years (36 eyes) had correction of +3.0 dsph, but it was not statistically significant. There was no statistically significant difference of the correction between pseudophakic (41) and phakic (19) eyes in patients younger than 59 years. The anterior movement of the IOL was 0.18 mm in the younger group and 0.15 mm in the older group. With good IOL movement and new materials which could have changeable refractive power, the problem of pseudophakic correction for near vision might be solved. PMID- 15794049 TI - [Prevalence of craniomandibular dysfunction signs and symptoms in students of the school of stomatology in Belgrade]. AB - Numerous epidemiological studies suggest that CMD problem is rather present in most countries all over the world, particularly in the Scandinavian and East European countries. There have not still been epidemiological studies on incidence of CMD in our surroundings, either organized or academic treatment programs of these diseases. The results of this study showed that most common CMD signs and symptoms were: impaired mandibular movements, deviation (deflection) in opening and closing the jaws, and TMJ sounds during mandible movements. Pain as a primary symptom was seen only in 5.2% of the analyzed subjects. Such fact indicates that this analyzed population group manifested the initial stage signs and symptoms of CMD. Recording of signs and symptoms mentioned above in young subjects should lead the therapist to perform more detailed analysis and apply adequate therapy modalities. PMID- 15794050 TI - [Complications of cochlear implant surgery]. AB - During the last several decades, cochlear implant has been fully recognized in treatment of severe hearing loss. Development of modern technology enabled inconceivable possibilities of technical qualities of the device as well as development of usable coding strategies, which led to extraordinary results in patient rehabilitation. Although cochlear implantation has become one of the routine operative procedures throughout the world nowadays, it gives rise to certain complications. These complications, though rare, can sometimes be very serious, even with fatal outcome. If cochlear implantation is performed by experienced and well-educated team of experts, the possibility of complications is minimal and is certainly not the argument against cochlear implantation as a method of treatment of severe hearing impairments. PMID- 15794051 TI - [Long-term results after elective and emergency surgery of abdominal aortic aneurysm]. AB - INTRODUCTION: Abdominal aortic aneurysm can be repaired by elective procedure while asymptomatic, or immediately when it is complicated--mostly due to rupture. Treating abdominal aneurysm electively, before it becomes urgent, has medical and economical reason. Today, the first month mortality after elective operations of the abdominal aorta aneurysm is less than 3%; on the other hand, significant mortality (25%-70%) has been recorded in patients operated immediately because of rupture of the abdominal aneurysm. In addition, the costs of elective surgical treatment are significantly lower. OBJECTIVE: The objective of this study is to compare long-term survival of patients that underwent elective or immediate repair of abdominal aortic aneurysm (due to rupture), and to find out the factors influencing the long-term survival of these patients. MATERIAL AND METHODS: Through retrospective review of prospectively collected data of the Institute for Cardiovascular Diseases of Clinical Center of Serbia, Belgrade, 56 patients that had elective surgery and 35 patients that underwent urgent operation due to rupture of abdominal aneurysm were followed up. Only the patients that survived 30 postoperative days were included in this review, and-were followed up (ranging from 2 to 126 months). Electively operated patients were followed during 58.82 months on the average (range 7 to 122), and urgently operated were followed over 52.26 months (range 2 to 126). There was no significant difference of the length of postoperative follow-up between these two groups. RESULTS: During this period, out of electively operated and immediately operated patients, 27 and 22 cases died, respectively. There was no significant difference (p>0.05a) of long-term survival between these two groups. Obesity and early postoperative complications significantly decreased long-term survival of both electively and immediately operated patients. Graft infection, ventral hernia, aneurysm of peripheral arteries and other vascular reconstructive procedures were the factors that significantly reduced long-term survival of patients operated immediately due to rupture. DISCUSSION: This comprehensive study has searched for more factors than others had done before. The applied discriminative analysis numerically evaluated the influence of any risk factor of mortality. These factors were divided in three groups as follows: preoperative, operative and postoperative ones. Preoperative factors were sex, age, diabetes mellitus, arterial hypertension, obesity, COPD, and naturally, the indication for operative treatment of ruptured or non-ruptured abdominal aneurysm. Among all these factors, only obesity significantly reduced long-term survival of electively operated patients. It may be said that immediately operated patients who survived the first 30 postoperative days had quite good long-term survival. Operative factors such as type of operative procedure and vascular graft had no influence on long-term survival of patients in both groups. Postoperative risk factors were early postoperative complications, graft infection, symptomatic cerebrovascular disease, carotid endarterectomy, myocardial revascularization, ventral hernias, "other" non vascular operations, malignancy, mental disorders, peripheral aneurysms and occlusive vascular disease, and other vascular operations either due to aneurysm or peripheral occlusive disease. Early postoperative complications (even graft infection) had no significant effect on long-term survival. Ventral hernias and peripheral aneurysms were factors that significantly decreased long-term survival of patients operated for rupture of the abdominal aneurysm. CONCLUSION: It is interesting that endarterectomy, myocardial revascularization or malignancy after repair of the abdominal aneurysm (ruptured or non-ruptured) had no effect on long-term survival. PMID- 15794052 TI - [Voiding dysfunction in children aged five to 15 years]. AB - Voiding dysfunction in children was analyzed in 91 patients in a period from January 1st to October 1st 1998. Most of the patients had functional voiding disorder (92.31%), and only 7.69% manifested monosymptomatic night enuresis. The number of girls was bigger in the group of patients with voiding dysfunction while the boys were predominant in the group with monosymptomatic nocturnal enuresis. More than a half of children with functional voiding disorder had repeated urinal infections (58.23%), incontinence (93.49%), need for urgent voiding (68.13%), and vesicoureteral reflux (47.61%). The most common type of voiding dysfunction was urge syndrome/urge incontinence. The incidence of dysfunctional voiding disorder was more often in children with scaring changes of kidney which were diagnosed by static scintigraphy. PMID- 15794053 TI - [Open wedge osteotomy and callus distraction by means of the external fixator in distal femur and proximal tibia in knee arthrosis with valgus and varus deformity]. AB - Various methods can be applied for treatment of knee deformity. One of them is open wedge osteotomy and callus distraction by means of the external fixator. This paper presents the results of treatment of 16 patients with knee arthrosis associated with varus and valgus deformities (10 varus and 6 valgus deformities). Open wedge osteotomy of the varus deformity was performed in the proximal tibia, and in case of the valgus deformity in the distal part of the femur. The patients who were operated on had a knee varus larger than 10 degrees and a knee valgus larger than 12 degrees. Prior to open wedge osteotomy and application of the external fixator, knee arthroscopy was performed (meniscectomy, cartilage drilling and shaving, debridement, the extraction of loose bodies). After one year follow-up, the final outcome of the treatment was positive in all patients. The treatment alleviated the pain in these patients. The method is minimally invasive and relatively easily applied. Mitkovic's external fixator type M20-CD-V allows for continuous callus distraction with simultaneous correction of the varus or valgus knee deformity. PMID- 15794054 TI - [Successful revascularization of the kidney--3 case reports]. AB - Three cases of successful kidney revascularization and recovery of renal function are presented in this study. In all three cases, renal failure and renovascular hypertension were caused by renal artery occlusion associated with aortic aneurysm (two abdominal and one thoracoabdominal). Prior to operation, one patient required dialysis 4 months, one 25 days and one 2 days. After kidney revascularization, renal function recovered immediately in the first case, in the second case after three months, and in the third case after 10 days. In one case, blood pressure restored to normal without medical therapy, while in two other cases blood pressure decreased nearly to normal with minimal medical therapy. In appropriately selected cases, revascularization of the occluded renal artery is recommended for treatment of both renal failure and renovascular hypertension. In such cases, collateral circulation is crucial to enable the preservation of dysfunctional kidney. PMID- 15794055 TI - [Floating spleen with chronic torsion of the pedicle causing splenomegaly and secondary hypersplenism]. AB - If the spleen is not fixed within the left subphrenic space, it gradually passes into the lower abdomen, where is much more exposed to trauma. Torsion of the splenic pedicle can also occur, causing the infarct necessitating an immediate surgery. Venous stasis causes splenomegaly and sometimes secondary hypersplenism. The authors present 16.5-year old girl with torsion of the splenic pedicle of floating spleen for 720 degrees: in spite of that, the patient had neither splenic infarct nor splenic vein thrombosis, possibly due to thrombocytopenia, but she had splenomegaly and secondary hypersplenism with pancytopenia causing bleeding, sideropenic anemia and mild jaundice. After treatment with iron, the patient underwent splenectomy which resulted in almost immediate rise of the number of all blood cells, and even thrombocytosis. The authors suggest early surgical treatment of the floating spleen, preferably splenopexy, before development of severe complications when splenectomy had to be performed in the majority of patients. Accessory spleens, if present, should be saved. PMID- 15794056 TI - [Ovarian cancer after in vitro fertilization]. AB - There is serious concern about cancer risk in women undergoing ovarian stimulation treatment for infertility and longterm safety of these procedures. Association between fertility drugs and ovarian cancer is still controversial. A 30-year-old woman was referred to our institution with the initial diagnosis of an adnexal tumor after in vitro fertilization. Her history revealed adnexectomy for mucinous cystadenofibroma of the left ovary eight years ago, and cystectomy due to cystadenoma of the right ovary three years ago. At admission, the most remarkable findings were high temperature and elevated white blood cell count and erythrocyte sedimentation rate. Broad spectrum antibiotic treatment was initiated. Six days after admission, the patient was feverish, with temperature up to 38 degrees C, and evident signs of the acute abdomen. Immediate laparotomy was performed and multilocular right adnexal tumor 150x130x100 millimeters in size was identified. The right salpingo-oophorectomy was carried out. Pathological diagnosis was mucinous ovarian adenocarcinoma. Eighteen days later, radical surgery was performed. After the surgery, chemotherapy was applied. There is an urgent need for clear interpretation of the association between fertility drugs and subsequent higher ovarian cancer risk. Lacking conclusive evidence, an increased risk of ovarian cancer has been reported and more recently disputed. Higher ovarian cancer risk may be serious and even life-threatening complication for women undergoing ovarian stimulation. PMID- 15794057 TI - [Mechanisms of protective effects of moderate alcohol consumption on the cardiovascular system]. AB - Consumption of alcoholic beverages has been known in many cultures since the ancient civilizations, so harmful consequences of excessive alcohol intake have been well explained. Many epidemiological studies confirmed lower morbidity and mortality rates of cardiovascular diseases in persons who drink alcohol "moderately" (1-2 drinks a day), independently of the kind of alcoholic beverage. This paper presents the review of molecular mechanisms that are believed to explain cardioprotective effect of moderate drinking--alcohol effects on lipoproteins, endothelial cells, blood clot formation and dissolution, as well as genetic and gender variances modifying the relation. A simple recommendation regarding the increase of alcohol consumption cannot be made on population level. The professionals must therefore concentrate on other preventive methods in order to reduce other significant risk factors of cardiovascular diseases. PMID- 15794058 TI - [Pathogenesis of Helicobacter pylori infection--bacterium and host relationship]. AB - Helicobacter pylori (H. pylori) colonizes the gastric mucosa of a half of the mankind. Duodenal ulcer is found in 15-25%, gastric ulcer in 13%, while gastric adenocarcinoma develops in 1% of all infected individuals. Pathogenesis of H. pylori infection is related to the virulence factors of the bacterium, environmental (dietary habits, hygiene, stress) and host factors (age, sex, blood type). Colonization of the gastric mucosa is related to the motility of the bacterium, presence of lipopolysaccharide (LPS) and various bacterial enzymes. Gastric mucosal injury is the result of H. pylori LPS, vacuolization cytotoxin (vacA), cytotoxin associated protein (cagA), heat shock proteins and factors responsible for neutrophil chemotaxis and activity. H. pylori colonizes the gastric mucosa and zones of ectopic gastric epithelium. H. pylori infection is transmitted via oral-oral, fecal-oral and iatrogenic way (during endoscopy). Higher prevalence of the infection is associated with lower socioeconomic level, lack of drinking water, and living in a community. Acute H. pylori gastritis is superficial pangastritis progressing into the chronic phase after 7-10 days. Gastric mucosal atrophy and intestinal metaplasia can develop during the course of H. pylori infection. Clearly defined factors that influence the outcome of H. pylori infection include bacterial strain, distribution of gastritis, acid secretion and gastric mucosal atrophy. PMID- 15794059 TI - [Current diagnosis and treatment of the osteoarticular tuberculosis]. AB - In 1993, tuberculosis was recognized as global medical problem by WHO. Recurring flare-up of this disease was triggered by difficult socioeconomic situation of the countries in transition, current development of immunocompromising diseases, and some other factors have contributed, too. The osteoarticular tuberculosis has been always postprimary and localized form of the disease, which, due to certain similarity to some other osteoarticular diseases, has been challenge for orthopaedic surgeons both from diagnostic and treatment aspects. Bacteriological diagnostics has had a limited value due to application of recommended tuberculostatic medication before obtaining specimens from bones or joints; and, on the other hand, rapid bacteriological methods, because of their expensive equipment, could be carried out in large mycobacteriology laboratories only. New visualizing methods, CT and MRI, have been particularly important for detection and monitoring of tuberculosis process in the spine during the early stage of the disease, what is impossible to achieve with native radiograms. Programmed administration of tuberculostatic drugs both in non-operative and surgical treatment has been proven necessary. PMID- 15794060 TI - [Management of idiopathic nephrotic syndrome in childhood]. AB - The management of idiopathic nephrotic syndrome (INS) in children includes immunosuppressive and symptomatic treatment. The response to corticosteroid therapy is the best prognostic marker of the disease. The majority of children with INS (about 85%) are steroid-sensitive as they normalize proteinuria within 4 weeks of daily, oral prednisone administration. The most of steroid-sensitive patients (94%) has minimal change of nephrotic syndrome, while the majority (80.5%-94.4%) of those who are steroid-resistant has focal segmental glomerulosclerosis or mesangioproliferative glomerulonephritis. Initial therapy of INS consists of 60 mg/m2/day prednisone daily for 4 weeks followed by 40 mg/m2 on alternate days for 4 weeks, thereafter decreasing alternate day therapy every month by 25% over the next 4 months. Thus, the overall duration of the initial corticosteroids course is 6 months that may be significantly protective against the future development of frequent relapses. Approximately 30% of patients experience only one attack and are cured after the first course of therapy; 10 20% have only 3 or 4 steroid-responsive episodes before permanent cure; the remaining 40-50% of patients are frequent relapsers, or steroid-dependent. Standard relapse therapy consists of 60 mg/m2/ day prednisone until urine is protein free for at least 3 days, followed by 40 mg/m2 on alternate days for 4 weeks. The treatment of frequent-relapses and steroid-dependent INS includes several different regimens: maintenance (6 months) alternate steroid therapy just above steroid threshold (0.1-0.5 mg/kg/48h), levamisole, alkylating agents (cyclophosphamide or chlorambucil) or cyclosporine. The worse prognosis is expected in steroid-resistant patients who are the most difficult to treat. Renal biopsy should be performed in them. At present, there is no consensus on therapeutic regimen for steroid-resistant patients. The following immunosuppressive drugs have been used with varying success: cyclosporine, intravenous methyl prednisone pulses alone or combined with or followed by alkylating agents, plasma-exchange, and angiotensin-converting enzyme (ACE) inhibitors. Symptomatic treatment includes: 1) dietary regimen with normal protein intake and salt restriction, 2) calcium and vitamin D are prescribed with steroids, 3) diuretics should be used in case of severe edema, 4) infusion of albumin in case of severe hypovolemia, 5) treatment of hypertension, 6) anticoagulant therapy, and 7) prophylactic antibiotics in high-risk patients. PMID- 15794061 TI - [Can outstanding research be done under less than ideal conditions?]. AB - Great scientific discoveries rarely originate from small and poor countries. However, the lives and achievements of three Yugoslav scientists who were active in the biomedical sciences, Laza K. Lazarevic (1851-1891), Ivan Djaja (1884 1957), and Pavao Stern (1913-1976), serve as an example of success in this environment. These scientists, as well as the majority of other successful investigators in small and poor countries, were trained in foreign and developed countries and, upon return, were given the freedom to start a selfdependent research program. They overcame many obstacles, including wars and civil unrests, to contribute significantly to certain medical fields. It is interesting that although a Jew, Stern was allowed to work during the World War II in Zagreb, which became capital of the so-called Independent State of Croatia, a puppet state under German control. Perhaps his good name among pharmacologists helped him to keep position during this tough period. Nowadays, new technologies requiring for biomedical research are rather expensive, and poor countries cannot afford to finance many scientists. Thus, selection of the most productive researchers is the challenge for those who finance scientific work. PMID- 15794063 TI - New roles for oncology nurses evolve beyond the bedside. PMID- 15794062 TI - [St. Luke and his cult as holy healer of the Serbs]. AB - Most school doctors, who lived in the period of Early Christianity from 1st to 4th century A.D. and who were canonized saints, have been known, up to these days, among people and in scientific and medical circles as Holy Healers. It is understood that only exclusively educated medical experts, trained to heal professionally and prepare medicines are considered Holy Healers. Out of all Holy Healers, St. Kosma and Damian, St. Panteleimon, St. Luke, etc., are highly respected by our people. St. Luke (1st century A.D.) is specially honored by Serbian nation. His relics were taken to Smederevo in 1453 and then the town became "the place of many cures and new healing spot". Out of these relics, only the foot of St. Luke was preserved in a very good condition and it remained in the possession of the Serbian Orthodox Church. In old documents written in old Greek and clerical-slavic language, St. Luke is glorified as "reliable doctor both for soul and body..." St. Luke is respected as a protector of medicine and pharmacy, doctors and pharmacists, and patients, as well as many families (family patron of the Serbs), even of the whole regions. Many chemist's shops and hospitals are named by this Saint, what is the confirmation that his cult and recognition of his personality and his work are still present in our milieu. PMID- 15794064 TI - Use of off-label drugs in cancer care again under review. PMID- 15794065 TI - Telenursing: nursing care without geographic boundaries? PMID- 15794066 TI - ONS helps secure fiscal year 2005 funding increases for nursing and cancer programs. PMID- 15794067 TI - Evidence of bacterial biofilms in a rabbit model of sinusitis. AB - BACKGROUND: Bacterial biofilms have been documented on middle ear mucosa, tonsils, and cholesteatoma. In addition, we have described previously bacterial biofilms on frontal recess stents placed during surgery for chronic sinusitis. We hypothesize that bacterial biofilms are present on the mucosa of animals with sinuses experimentally infected with Pseudomonas aeruginosa. METHODS: We instilled Pseudomonas aeruginosa at log phase growth into the right maxillary sinus in 22 New Zealand white rabbits and harvested specimens 1-20 days later. We then examined the tissues with scanning electron microscopy (SEM) and light microscopy. Mucosa from the middle turbinate on the contralateral side was used as a control. RESULTS: We identified evidence of bacterial biofilms on the mucosa of all 22 animals under SEM. Twenty-one of 22 animals had sinus cultures positive for Pseudomonas. Bacterial biofilms were identified by evidence of glycocalyx, water channels, and three-dimensional structure. These images were similar to other images of known biofilms. CONCLUSION: This is evidence of the presence of bacterial biofilms in an animal model of sinusitis. This model may serve as a means to study the efficacy and safety of pharmacologic and surgical treatments on the disruption and elimination of bacterial biofilms. Additional study into the role of bacterial biofilms in perpetuating chronic sinusitis is warranted. PMID- 15794068 TI - Expression, localization, and significance of vascular permeability/vascular endothelial growth factor in nasal polyps. AB - BACKGROUND: The exact etiologic mechanisms leading to the formation of nasal polyps have remained largely obscure. A key phenomenon of this specific type of chronic inflammatory disease in nasal respiratory mucosa is remarkable edema. Vascular permeability/vascular endothelial growth factor (VPF/VEGF) plays an important role in inducing angiogenesis and modulating capillary permeability. OBJECTIVE: To study the expression and localization of VPF/ VEGF as a putative key factor in nasal polyp development. METHODS: Specimens of nasal polyps (n = 12) were harvested during endonasal sinus surgery in patients with polypous chronic rhinosinusitis. Specimens of healthy nasal respiratory mucosa (n = 12) served as controls and were obtained from inferior turbinates of patients undergoing surgery for nasal obstruction without signs and symptoms of inflammatory disease. Frozen sections were immunohistochemically stained for VPF/VEGF and quantitatively analyzed, using computer-based image analysis. RESULTS: The expression of VPF/VEGF in specimens of nasal polyps was significantly stronger than in specimens of healthy nasal mucosa of controls. VPF/VEGF in polypous tissue was mainly localized in vascular endothelial cells, in basal membranes and perivascular spaces, and in epithelial cells. CONCLUSION: The markedly increased expression in nasal polyps as opposed to healthy nasal mucosa suggests that VPF/VEGF may play a significant role in both the formation of nasal polyps and in the induction of heavy tissue edema. This finding is discussed with respect to the differential expression of cyclooxygenase (COX) isoenzymes-1 and -2 (COX-1 and COX-2) in nasal polyps was significantly stronger than in specimens of healthy nasal mucosa of controls. VPF/VEGF in polypous tissue was mainly localized in vascular endothelial cells, in basal membranes and perivascular spaces, and in epithelial cells. Conclusion: The markedly increased expression in nasal polyps as opposed to healthy nasal mucosa suggests that VPF/VEGF may play a significant role in both the formation of nasal polyps and in the induction of heavy tissue edema. This finding is discussed with respect to the differential expression of cyclooxygenase (COX) isoenzymes-1 and -2 (COX-1 and COX-2) in nasal polyps. PMID- 15794069 TI - The role of RANTES in nasal polyposis. AB - BACKGROUND: Characteristic infiltrates of eosinophils are a hallmark of nasal polyps (NPs). Several studies suggest that members of the CC chemokine family may be involved in this process. RANTES (regulated on activation, normal t-cell expressed and secreted) is a member of the CC chemokine family with chemotactic activity on mainly eosinophils and T lymphocytes. Thus, RANTES is an interesting target for the recruitment of eosinophils and T lymphocytes into the nose. The degree of the tissue eosinophilia has been reported to correlate with the severeness of the symptomatology of the disease and the extension on the lower respiratory tract, as well as with the probability of the recurrence of NPs. Therefore, we hypothesized that high numbers of eosinophils correlate with high levels of RANTES and that associated atopic diseases modify this correlation. METHODS: Total RNA was extracted from NP homogenates, reverse transcribed and RANTES mRNA expression analyzed using semiquantitative reverse transcription polymerase chain reaction and Northern blot analysis. Histological studies divided NPs in an eosinophilic and low eosinophilic group. Additionally, RANTES protein concentration was measured in homogenates by a RANTES-specific enzyme linked immunosorbent assay. RESULTS: This study has clearly shown that RANTES is expressed and secreted in NPs. The group with a high tissue eosinophilia had a significant higher RANTES gene expression and protein production than NPs without tissue eosinophilia. The isolated coincidence of acetylsalicyl acid intolerance with chronic hyperplastic sinusitis/NP additionally increased significantly the RANTES amounts in NPs. CONCLUSION: Increased RANTES leads to increased tissue eosinophilia. Associated acetylsalicylic acid intolerance seems to enhance the amount of RANTES in NPs and might explain in part the more severe clinical course in those patients. Thus, RANTES appears to play an important role in mobilization of eosinophils into the local inflamed tissue. PMID- 15794070 TI - An assessment for the presence of bacterial contamination of Venturi principle atomizers in a clinical setting. AB - BACKGROUND: Venturi principle atomizers have been criticized for potential contamination and disease transmission when used for nasal application. A recent laboratory study reported a high rate of contamination of these atomizers after their tips were immersed and sprayed into a broth containing Staphylococcus aureus. The objective of this study was to determine whether the Venturi-type atomizers used regularly at a busy referral center for sinonasal disorders were contaminated with bacteria. METHODS: Fifteen Venturi-type atomizers containing 2% lidocaine (four bottles), 4% lidocaine (three bottles), or 0.05% oxymetazoline (eight bottles) were sprayed onto blood agar plates. All atomizer bottles had been in routine use in a tertiary rhinology clinic for 1 month. The contents within each bottle's reservoir also were cultured. Thus, the total sample size was 30 plates. Then, the plates were incubated at 35 degrees C for 48 hours and then at room temperature for 72 hours. RESULTS: Three of our 30 plates grew one colony each of Gram-positive bacilli, Gram-positive cocci, and Staphylococcus species. CONCLUSION: The rare growth of any bacteria seen in our samples could as likely represent contamination during processing and handling of the plates as contamination of the atomizers themselves. Thus, Venturi-type atomizers may not have a high potential contamination risk, which has been suggested by previous reports. PMID- 15794071 TI - Efficacy and safety of single and multiple doses of pseudoephedrine in the treatment of nasal congestion associated with common cold. AB - BACKGROUND: Pseudoephedrine (60 mg) is widely used as an oral decongestant taken in tablet or syrup formulations every 4-6 hours for the treatment of nasal congestion associated with common cold and allergy. However, there are relatively few studies in the literature that have used objective measures of nasal airway resistance (NAR) to assess the efficacy of pseudoephedrine, and most studies use only a single dose of medication. The present study has the aims of studying the safety and efficacy of a new pseudoephedrine formulation after single and multiple doses in patients with URTI. METHODS: The study was a double-blind, randomized, parallel-group, placebo-controlled trial conducted over three study days at a single center. Patients suffering from nasal congestion associated with common cold were recruited and total NAR was measured by the technique of posterior rhinomanometry. NAR and subjective scores of nasal congestion were measured at baseline and after dosing with study medication, every hour over a four-hour period on day 1 after a single dose, and on day 3 after multiple doses of medication. Subjective scores of congestion/stuffiness were also made as a summary score at the end of each day of treatment. RESULTS: Two hundred and thirty-eight patients with nasal congestion associated with acute upper respiratory tract infection (URTI), mean age 20 years, were recruited to the study and received treatment. After a single dose on day 1 the pseudoephedrine group had a statistically significant lower area under the NAR curve than placebo (p = 0.006) for the primary efficacy variable area under the NAR curve from 0-3 hours (NAR AUC 0-3h), and similarly for the secondary efficacy variable NAR AUC 0 4h (p = 0.001). On day three after multiple doses, the pseudoephedrine group had a statistically significant lower NAR AUC 0-3h and AUC 0-4h than placebo (p < 0.001), On day 1, the pseudoephedrine group had significantly lower subjective scores for congestion than placebo visual analog scale (VAS) AUC 0-3h (p = 0.029) and similarly for VAS AUC 0-4h (p = 0.021). On day 3, the differences in subjective scores were not significantly different. The mean decrease from baseline of the summary score for congestion/stuffiness over the duration of the study was greater in the pseudoephedrine group compared to the placebo group (p = 0.016). On average, heart rate was between two and four beats per minute greater in the pseudoephedrine group compared to placebo. Five adverse events were reported in both treatment groups and these were deemed to be unrelated to treatment. CONCLUSION: The results demonstrate that pseudoephedrine is a safe and effective treatment for nasal congestion associated with URTI. The results from the laboratory study on day 1 demonstrate by both objective and subjective measures of nasal congestion that a single dose of 60 mg pseudoephedrine is superior to placebo treatment. Support for the decongestant efficacy of multiple doses of pseudoephedrine is provided by objective measures on day 3 and subjective measures made over three days, but not by the VAS scores on day 3. PMID- 15794072 TI - Nasal obstruction and sleep-disordered breathing: a study using acoustic rhinometry. AB - BACKGROUND: The relationship between nasal airway function and sleep-disordered breathing (SDB) remains unclear. Although correction of nasal obstruction can significantly improve nighttime breathing in some patients, nasal obstruction may not play a role in all cases of SDB. An effective method of stratifying these patients is needed. Acoustic rhinometry (AR) is a reliable, noninvasive method of measuring the dimensions of the nasal airway. METHODS: In 44 patients, we performed acoustic rhinometric measurements of nasal airway cross-sectional area, followed by hospital-based polysomnography and nasal continuous positive airway pressure (nCPAP) level titration. We compared anatomic nasal obstruction to perceived nasal obstruction, as well as respiratory distress index and nCPAP titration level, using the Pearson correlation and multiple linear regression analysis within body mass index groups. RESULTS: Perceived nasal obstruction correlated significantly with objective anatomic obstruction as measured by AR (r = 0.45, p < 0.01). For certain subgroup analyses in patients with a body mass index below 25, AR measurements correlated significantly with both nCPAP titration pressure (r = 0.85, p < 0.01) and respiratory distress index (r = 0.67, p = 0,03). CONCLUSION: Nasal airway function may be a significant component of SDB in some patients, perhaps playing a larger role in patients who are not overweight. The best responders to nasal surgery for SDB may be nonoverweight patients with nasal obstruction. AR along with nasal examination may be helpful in the evaluation and treatment of the SDB patient. PMID- 15794073 TI - Effect of using multiple culture media for the diagnosis of noninvasive fungal sinusitis. AB - BACKGROUND: This study was designed to assess the relative efficiency of three different culture media for isolating fungi in patients suspected of having noninvasive fungal sinusitis. METHODS: A prospective study was performed of 209 operative samples of sinus "fungal-like" mucin from 134 patients on 171 occasions and processed for microscopy and fungal culture in Sabouraud's dextrose agar, potato dextrose agar, and broth media. RESULTS: Ninety-three (69%) of 134 patients had evidence of fungal infection. Two-thirds of patients had negative microscopy samples yet 56% of these went on to positive cultures. Forty-five percent cultured Aspergillus genus. Discrepancy between the fungi cultured in different media and on different occasions was common. With a single culture medium up to 19% of patients and 15% of samples would have been falsely labeled fungal negative. CONCLUSION: Increasing the number and type of fungal culture media used increases the number and range of fungal isolates from mucin in patients with the features of fungal sinusitis. Negative specimen microscopy is unreliable. All specimens should be cultured in multiple media and on multiple occasions when fungal sinusitis is suspected. PMID- 15794074 TI - Presence of fungus in sinus cultures of cystic fibrosis patients. AB - BACKGROUND: Sinonasal pathology is nearly universal in the cystic fibrosis (CF) population. The bacteriology of sinus cultures from CF patients and the implications of sinus bacterial pathogens in this group have been studied; however, sinus fungal isolates from CF patients have not been examined in the literature. METHODS: We reviewed 30 consecutive CF patients undergoing endoscopic sinus surgery at our institution for the presence of fungal isolates obtained from the sinuses at the time of surgery. RESULTS: Thirty-three percent of fungal cultures were positive in this sample; in addition, two patients were newly diagnosed with allergic fungal sinusitis. CONCLUSION: We examine the possible implications of positive fungal sinus cultures in the CF population. PMID- 15794075 TI - Long-term rhinoflowmetry: a new method for functional rhinologic diagnostics. AB - BACKGROUND: With current functional diagnostic tools in rhinology (rhinomanometry, rhinoresistometry, and acoustic rhinometry) long-term assessment of nasal function is difficult to acquire. Usually, only the situation at the time of examination is evaluated. Therefore, temporary nasal complaints of the nasal cycle are difficult to assess. It was the aim of this work to create a diagnostic tool to measure nasal flow over a long time period under physiological and everyday life conditions. We term the method long-term rhinoflowmetry (LRFM). METHODS: A portable device recording nasal airflow over a time period up to 72 hours was developed. Kinetic pressure fluctuations during respiration as a measure for the flow were registered over time and relative airflow was calculated. A summary of diagnostic results is given in six exemplary patients. RESULTS: Classic nasal cycles could be recorded in detail with durations ranging from 90 minutes to 10 hours. The manifestation of nighttime nasal obstruction as well as a case of paradoxical nasal obstruction were objectified. LRFM enables the assessment of time, duration, and extent of temporary nasal obstruction. CONCLUSION: LRFM enables an assessment of temporary nasal obstruction as well as physiological and pathological fluctuations in the nasal cycle. Especially in cases in which traditional rhinological diagnostic tools are unsatisfying, the enhanced diagnostic quality of LRFM appears to be a promising supplement to the currently available rhinological monitoring methods. PMID- 15794076 TI - Decreased connexin 43 expression correlated with eosinophil infiltration in nasal polyps. AB - BACKGROUND: A nasal polyp usually is characterized by eosinophil infiltration. Eosinophil-fibroblast interaction is an important event of persistent inflammation in airways. We have found abundant connexin 43 (Cx43) expression in subepithelial fibroblasts of nasal mucosa. Thus, we aim to analyze the relationship of Cx43 expression and eosinophil in nasal polyps. METHODS: In 25 nasal polyps and 19 inferior turbinates, indirect immunofluorescent and hematoxylin and eosin staining were performed in adjacent sections. We calculated the density of Cx43 staining and eosinophil individually by fluorescent and light microscope. RESULTS: Positive Cx43 staining under confocal microscope was shown as punctated spots on cell margin. The density of Cx43 and eosinophil staining was significantly different between groups of inferior turbinate and nasal polyp (p = 0.01 and 0.03, respectively). Decreased Cx43 expression in the subepithelial fibroblast was correlated with eosinophil infiltration in nasal polyps. Spearman rank order coefficient was equal to -0.43 (p < 0.05). CONCLUSION: This is the first demonstration of decreased Cx43 expression related to eosinophil infiltration. To the best of our knowledge, interleukin-8 may be a link between Cx43 and eosinophil and orchestrating both in developing nasal polyps. PMID- 15794077 TI - Morphology and distribution of nasal telangiectasia in HHT-patients with epistaxis. AB - BACKGROUND: Epistaxis in hereditary hemorrhagic telangiectasia (HHT; Rendu-Osler Weber syndrome) is a frequent symptom that may be caused by a multitude of different genetic and epigenetic phenomena. This investigation analyzes the distribution of nasal telangiectasia in 21 patients with HHT. METHODS: The patients were examined for endonasal telangiectasia by videoendoscopy with rigid endoscopes; in addition, the anterior portion of the nose was examined under the operating microscope. The endonasal findings were recorded on videotape and then evaluated in the media laboratory. RESULTS: Morphology of the nasal telangiectasia showed wide variations: the vessels were shaped like spots, loops, or spiders or they clustered and resembled raspberries. Gender did not have an influence on the phenotype of telangiectasia, whereas advancing age correlated with a higher density of telangiectasia. Patients with an intact nasal septum exhibited the bulk of telangiectasia in the anterior nasal cavity but also on the middle turbinates, the floor of the nose, and within the valve area. Patients with septal perforations displayed the majority of telangiectasia around the edge of the perforations, on the floor of the nose, and on the turbinates. Scattered telangiectasia also could be found in the profound parts of the nasal cavity and in the nasopharynx, especially in patients with septal perforations. CONCLUSION: The shapes of endonasal telangiectasia in HHT patients are very heterogeneous; predilection sites could first of all be found within the anterior portion of the nose. Morphology and distribution of endonasal telangiectasia change as a result of therapeutic interventions, development of septal perforations, and with advancing age. Therefore, repeat endoscopies are recommended to assess the actual stage of the disease before epistaxis therapy. PMID- 15794078 TI - The SF-36 health status questionnaire in assessing patients with epistaxis secondary to hereditary hemorrhagic telangiectasia. AB - BACKGROUND: This study assesses the ability of the short form 36 (SF-36), a validated health status survey, to measure the health of patients with epistaxis due to hereditary hemorrhagic telangiectasia (HHT). METHODS: Thirty-eight patients completed the SF-36 and symptom-specific questionnaires. They were asked to rate their epistaxis as severe, moderate, or mild. The SF-36 data are compared with a reference population and analyzed with respect to the subgroups. RESULTS: The scores for each dimension of health were significantly reduced (p < 0.05) when compared with the reference population for all dimensions except pain. When compared with the patients' ratings of severity, a significant correlation was detected in five dimensions (p < 0.05). CONCLUSION: The SF-36 reflects the reduced health status of patients with epistaxis due to HHT. Changes in the SF-36 score could be used as an outcome measure in assessing efficacy of treatment of this condition. PMID- 15794079 TI - The effect of treatment for epistaxis secondary to hereditary hemorrhagic telangiectasia. AB - BACKGROUND: Epistaxis is the most common symptom in patients with hereditary hemorrhagic telangiectasia. Different treatment options have been described but with little reference to their effect on quality of life or disease severity. METHODS: This study prospectively investigated the effect of nasal closure, argon laser, and a combination of septodermoplasty and argon laser treatments on quality-of-life scores and disease severity ratings in a group of 29 patients with moderate to severe epistaxis referred to a tertiary referral center. RESULTS: The only group of patients who showed a significant improvement (p < 0.05) in quality of life were those who underwent nasal closure. In addition, the nasal closure patients were the only group in which there was a complete cessation of bleeding. CONCLUSION: Nasal closure should be offered to patients with moderate to severe epistaxis secondary to hereditary hemorrhagic telangiectasia that has proved unresponsive to other treatment. PMID- 15794080 TI - Economic analysis of the treatment of posterior epistaxis. AB - BACKGROUND: The aim of this study was to compare the economic impact of controlling posterior epistaxis by transnasal endoscopic sphenopalatine artery ligation (TESPAL) and endovascular embolization. METHODS: We conducted a retrospective chart review of patients undergoing treatment of posterior epistaxis by either TESPAL or embolization. International Classification of Diseases 9 code 784.7 (epistaxis) was the initial screen followed by common procedural terminology codes for TESPAL and angiography with embolization. The total charges and direct costs for TESPAL and endovascular embolization were determined. An unpaired Student's t-test was used to evaluate statistical significance. RESULTS: Analysis revealed 25 patients that met inclusion criteria. The mean total charge was $14,088 for embolization and $7561 for TESPAL. The differences were statistically significant (p < 0.00006). Costs, defined as reimbursement by third-party payers and direct payments, varied widely and their difference did not reach statistical significance in this sample. CONCLUSION: Our data established no economic advantage for angiography and, in fact, show a trend toward this treatment being more expensive than TESPAL. TESPAL is a procedure that can be performed quickly and on an outpatient basis without the need for angiography equipment or expertise. Additionally, the procedure provides the advantage of a comprehensive endoscopic nasal evaluation for ruling out tumors or other intranasal lesions. With equal efficacy, at least equal costs and equal risk, and additional diagnostic advantages, TESPAL is a more rational treatment for posterior epistaxis. PMID- 15794081 TI - Endoscopic biopsy of human olfactory epithelium as a source of progenitor cells. AB - BACKGROUND: The adult central nervous system contains progenitor cells; however, invasive surgery is required for their harvest. Olfactory neuroepithelium (ONe) has attracted attention because it is extracranial and contains progenitor cells that account for its regenerative capacity. Olfactory progenitor cells have been cultured from postmortem ONe. Our aim was to determine if olfactory progenitors could be obtained via biopsy from patients in a feasible, effective, and safe manner. METHODS: Endoscopic biopsy was performed on individuals undergoing sinus surgery (n = 42). Olfactory function was assessed pre- and postoperatively. Specimens were cultured under conditions for olfactory progenitor cell development. RESULTS: Progenitor cells emerged in cultures from 50% of our patients. The superior turbinate, biopsied with cutting punch forceps, gave the highest yield. No adverse impact on olfaction or complications with the biopsy were observed. CONCLUSION: Endoscopic biopsy of ONe for obtaining olfactory progenitor cells from living donors is feasible, effective, and safe. PMID- 15794082 TI - Correlation between preoperative symptom scores, quality-of-life questionnaires, and staging with computed tomography in patients with chronic rhinosinusitis. AB - BACKGROUND: This study evaluates the correlation between preoperative symptoms, quality-of-life questionnaires (chronic sinusitis survey and 20-Item Sinonasal Outcome Test [SNOT-20]), and staging on computer tomography (CT). It consisted of a prospective cohort study of all consecutive patients undergoing surgery for medically nonresponsive chronic sinusitis, which took place at a tertiary care center. METHODS: Two hundred twenty-one patients completed the Chronic Sinusitis Survey (CSS) questionnaire and the SNOT-20. A visual analog scale (VAS) symptom score was completed also. The average age of the patients was 44.5 years and the male/female ratio was 1.3:1. The symptom scores of all three questionnaires were compared with the Lund-MacKay CT scan score of the sinuses. RESULTS: The median Lund-MacKay CT score was 12 (mean +/- SD, 12.7 +/- 5.7). There was no significant correlation between the SNOT-20 questionnaire and the Lund-MacKay CT score (p = 0.026; p = 0.764) and between the CSS and the Lund-MacKay CT score (p = -0.158; p = 0.058). Furthermore, there was no significant correlation between a single VAS symptom score relating to overall sinonasal symptom severity and the Lund-MacKay CT scan score (p = 0.135; p = 0.121). However, a weak but statistically significant correlation was found between the VAS score based on the sum of five sinonasal symptoms and the Lund-MacKay CT scan score (p = 0.197; p = 0.020). CONCLUSION: The score, based on the sum of five sinonasal VAS symptoms, correlates to the disease severity as measured by the Lund-MacKay CT scan score. The SNOT-20 questionnaire and the CSS symptom score do not correlate to the Lund MacKay CT scan score, although a correlation was found between the CSS score and the Lund-MacKay CT score in various sub-groups of our patient population. PMID- 15794083 TI - Glycoconjugate expression in normal human inferior turbinate mucosa: a lectin histochemical study. AB - BACKGROUND: Further study on the composition and concentration of normal nasal mucosal glycoconjugates is needed. METHODS: A battery of ten lectins was used to stain six carbohydrates residing in the inferior turbinate mucosa of 21 healthy individuals. RESULTS: The concentrations of glycoconjugates of the glycocalyx exceeded that of epithelial goblet, ciliated, and basal cells, and also of mucous, serous, and mixed submucosal glands. Epithelial goblet cells and submucosal mucous glands contained relatively high concentrations of glycoconjugates, whereas in epithelial ciliated and basal cells, similar to serous and mixed submucosal glands, concentrations were scanty. The concentrations of N-acetylgalactosamine and galactose were higher in goblet cells than in mucous glands (p = 0.006, and p = 0.056, respectively). Differences in the concentrations of N-acetylglucosamine, fucose, mannose, and sialic acid were insignificant. CONCLUSIONS: The findings provide a baseline for comparison of the composition and concentration of carbohydrates of the normal and pathologic inferior turbinate mucosa. PMID- 15794085 TI - [Determination of dl-alpha-tocopherol acetate and dl-alpha-tocopherol in foods by HPLC using post-column photochemical reaction]. AB - A reliable analytical method for the simultaneous determination of dl-alpha tocopherol acetate and dl-alpha-tocopherol in foods was established by HPLC using post-column photochemical reaction with UV and fluorescence detection. For low fat food such as fruit juice and vegetable sauce, the tocopherols were extracted with methanol containing 0.1% ascorbic acid and the extract solution was injected into the HPLC. For fatty foods such as butter and margarine, the tocopherols were extracted with a mixed solvent of acetonitrile-2-propanol (9:1) containing ascorbic acid. The extract was cleaned up using a Sep Pak plus C18 cartridge and the eluent from the cartridge was injected into the HPLC. The peaks corresponding to tocopherols on the chromatogram were confirmed by comparing their UV spectra with those of the standard mixture at lamp-on and lamp-off of the photochemical reactor. The recoveries of tocopherols from low-fat foods (orange juice and barbecue sauce) fortified at levels of 10 and 100 microg/kg each were 88.3 to 105.8% (RSD 0.5 to 6.0%) and those from the fatty foods (peanut butter and margarine) fortified at 100 microg/kg each were 57.1 to 88.3% (RSD 3.0 to 6.4%). The determination limits corresponded to 10 microg/kg of the tocopherols in the low-fat foods and 20 microg/kg in the fatty foods. PMID- 15794084 TI - Choanal stenosis: an unusual late complication of radiation therapy for nasopharyngeal carcinoma. AB - BACKGROUND: Choanal stenosis, usually a congenital anomaly in children, recently has been recognized as a late complication of radiation therapy (RT) for nasopharyngeal carcinoma (NPC). METHODS: We described the clinical history, preoperative evaluation, surgical management, and postoperative course of a case of acquired choanal stenosis after RT. RESULTS: The patient, a 39-year-old woman, presented with a history of NPC 16 years before presentation that had been successfully treated with RT On presentation, the patient complained of decreased nasal airflow. Bilateral choanal stenosis was confirmed per rigid nasal endoscopy. Transnasal endoscopic repair with mitomycin application was performed, and nasal stents were left in place for 6 weeks. Postoperative endoscopic examination showed patent choanae and a patent nasopharynx without stenosis. The patient continues to have good airflow 20 months postoperatively. CONCLUSION: Choanal stenosis can occur as an unusual complication of RT for NPC secondary to postradiation fibrosis. This is the first report of such a complication in the West. PMID- 15794086 TI - [Effects of various ginkgo biloba extracts and proanthocyanidin on hepatic cytochrome P450 activity in rats]. AB - In previous papers, we showed that Ginkgo biloba extract (GBE) induced hepatic cytochrome P450 (CYP) activity, in particular pentoxyresorufin O-dealkylase (PROD; corresponding to CYP2B type) in rats, and that GBE influenced the efficacy of co-administered drugs. In this study, to clarify the nature of the induction, we examined the effects of GBE samples from different sources and some major constituents of GBE on rat hepatic CYP in vitro and in vivo. In the study in vitro, eight GBE samples dose-dependently inhibited PROD activity in microsomes prepared from GBE-treated rats, and the inhibitory ratio correlated well with the content of proanthocyanidin in the GBE samples. Moreover, among six GBE constituents examined, proanthocyanidin markedly inhibited the PROD activity. However, administration of two GBE extracts with different proanthocyanidin contents to rats induced hepatic CYP activity, including PROD, to similar extents, and proanthocyanidin alone did not induce PROD activity. Furthermore, GBE samples extracted with both acetone-water and ethanol-water showed similar induction of CYPs in rats in vivo. These results suggest that most GBE samples available in Japan induce CYPs in rats regardless of the preparation method of the GBE, and that proanthocyanidin is not responsible for the induction. Further studies will be necessary to identify the constituent(s) of GBE involved in the induction of CYPs in vivo. PMID- 15794087 TI - [Analysis of residual solvents in natural flavorings by headspace GC using the standard addition method]. AB - Headspace GC using the standard addition method has been developed for the simultaneous determination of organic solvents in natural flavorings. The procedure can be outlined as follows: an aliquot of the sample is transferred to a 10 mL vial. To each vial, a DMSO solution containing solvents at different concentrations is added as the standard solution. The vials are kept at 50 degrees C (for automatic injection) or 40 degrees C (for hand-operated injection) for 40 minutes. One mL of the vapor phase in each vial is injected into a gas chromatograph equipped with an Aquatic-2 column (0.25 mm i.d. x 60 m). To evaluate this method, we conducted a performance study in collaboration with 10 laboratories, using ginger oleoresin. We analyzed 6 solvents (methanol, 2 propanol, acetone, dichloromethane, hexane, and 1,1,2-trichloroethene) for which the maximum residue limits are established in Japan's Specifications and Standards for Food Additives. Methanol and acetone existed in the ginger oleoresin, so only the other that four kinds of solvents were added to it. Eight of the laboratories used automatic injection, while the remaining two used hand operated injection. Statistical analyses were conducted on the data obtained from the 8 laboratories. Repeatability standard deviations (RSDr) ranged from 4.3 to 11.4%, and reproducibility standard deviations (RSDR) ranged from 8.4 to 19.0%. PMID- 15794088 TI - [Analysis of trichothecenes in barley tea and beer by liquid chromatography/tandem mass spectrometry]. AB - A simple method for analysis of trichothecenes [Type A: diacetoxyscirpenol, neosolaniol, HT-2 toxin, and T-2 toxin, Type B: deoxynivalenol, nivalenol, fusarenon-X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivarenol] in barley tea and beer using liquid chromatography tandem mass spectrometry (LC/MS/MS) was developed. Trichothecenes were extracted with ethyl acetate-methanol (19:1). The solvent was evaporated to dryness and the residue was dissolved in water-methanol (3:1) for injection into the LC/MS/MS. The LC separation was performed with an octadecylated silica column at a flow-rate of 0.2 mL/min, using a mobile phase consisting of water, methanol and acetonitrile. MS/MS was used in multiple reaction monitoring, employing electrospray ionization (ESI-MRM). The recoveries of trichothecenes from drinks at 1 microg/L (Type A) and 10 microg/L (Type B) were 52.5-115.2% (barley tea) and 68.1-127.5% (beer). Five barley tea and ten beer samples were analyzed by this method. Trichothecenes were not detected in them. This method may have applications in quality assurance. PMID- 15794089 TI - [Inter-laboratory evaluation studies for establishment of notified ELISA methods for allergic substances (buckwheat)]. AB - Inter-laboratory evaluation studies were conducted for the ELISA methods for allergic substances (buckwheat). Extracts of snack, bun and udon spiked with buckwheat standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate at 10 laboratories. Coefficients of variation (CVs) of the ELISA methods using a Buckwheat Protein ELISA Kit (Buckwheat kit) and a FASTKIT Buckwheat ELISA kit (Buckwheat ELISA kit) were mostly below 10%. Mean recoveries of the buckwheat standard protein from the food extracts were over 40% in the two ELISA methods. Repeatability relative standard deviations of buckwheat standard protein in three food extracts were in the ranges of 6.8-78.5% and 5.0-33.9% for the Buckwheat kit and the Buckwheat ELISA kit, respectively. Reproducibility relative standard deviations of buckwheat standard protein in three food extracts were 11.9-69.5% and 16.5-34.1% for the Buckwheat kit and the Buckwheat ELISA kit, respectively. The detection limits of both ELISA methods were 1 ng/mL in sample solutions. These results suggest that the notified ELISA methods are reliable and reproducible for the inspection of buckwheat protein levels in extracts of snack, bun and udon. PMID- 15794090 TI - [Analysis of pesticides including chlorine in welsh onions and mushrooms using gas chromatograph with an atomic emission detector (GC-AED)]. AB - An analytical method for the determination of 32 kinds of pesticide residues in onions, Welsh onions and mushrooms using gas chromatograph with an atomic emission detector (GC-AED) was developed. The pesticides were extracted with acetone-n-hexane (2:3) mixture. The crude extract was partitioned between 5% sodium chloride and ethyl acetate-n-hexane (1:4) mixture. The extract was passed through a Florisil mini-column for cleanup with 10 mL of acetone-n-hexane (1:9) mixture. Although the sensitivity of GC-AED was inferior to that of GC-ECD, GC AED has a superior element-selectivity. Therefore pesticide residues in foods could be analyzed more exactly by using GC-AED. Thirty-two pesticides including chlorine in onion, Welsh onion and shiitake mushroom were detected without interference. Recoveries of these pesticides from samples determined by GC-AED were 64-114%, except for a few pesticides. PMID- 15794091 TI - [Inter-laboratory evaluation studies for establishment of notified ELISA methods for allergic substances (peanuts)]. AB - Inter-laboratory evaluation studies were conducted for ELISA methods for allergic substances (peanuts). Extracts of biscuit, sauce, chocolate and butter spiked with peanut standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of the ELISA methods using a Peanut Protein ELISA Kit (Peanut kit) and a FASTKIT Peanut ELISA kit (Peanut ELISA kit) were mostly below 10%. Mean recoveries of the peanut standard protein from the food extracts were over 40% in the two ELISA methods. Repeatability relative standard deviations of peanut standard protein in four food extracts were in the ranges of 15.2-49.7% and 3.0-28.3% for the Peanut kit and the Peanut ELISA kit, respectively. Reproducibility relative standard deviations of peanut standard protein in four food extracts were 23.5-44.4%, 9.6 28.4% for the Peanut kit and the Peanut ELISA kit, respectively. The detection limits of both ELISA methods were 2-2.5 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of peanut protein levels in extracts of biscuit, sauce, chocolate and butter. PMID- 15794092 TI - [The problem and improvement of colorimetric method for determination of sulfite in foods containing sulfur compounds]. AB - The modified Rankine colorimetric method for measuring sulfite added to food as a food additive has a low determination limit and is little influenced by interfering substances from foods. However, it can give erroneous results for foods containing Liliaceae Allium. So, four different methods, alkaline titration, a colorimetric method, ion chromatography and qualitative analysis with potassium iodate-starch paper, were examined. It was found that the sodium azide used in the colorimetric method forms sulfur dioxide during bubbling and heating. The proposed colorimetric method can be applied to food containing sulfur compounds, if sodium azide is omitted and 1% triethanolamine solution is used as an absorbent instead of 0.1 mol/L sodium hydroxide solution. PMID- 15794093 TI - [Estimation of daily intake of phenols in hospital meal samples]. AB - Daily intakes of 12 phenols which are possible endocrine disruptors were estimated in hospital meals from 2000 to 2001. 4-Nonylphenol (4-NP (mix)) and bisphenol A (BPA) were detected at levels of 5.0 to 19.4 ng/g and 0.2 to 1.1 ng/g, respectively. 4-tert-Butylphenol, 4-n-pentylphenol, 4 -tert-pentylphenol, 4 n-hexylphenol, 4-n-heptylphenol and 4-tert-octylphenol were detected at levels of 0.1 to 2.4 microg/g. The daily intakes of 4-NP (mix) and BPA were 5.8 microg/day and 0.42 microg/day, respectively. The daily intakes of other phenols were less than 1 microg/day. PMID- 15794094 TI - [Relation between food allergy and dietary life]. PMID- 15794095 TI - [National control of avian influenza and management in disease prevention]. PMID- 15794096 TI - [Revival of a traditional food, puffer liver "fugu kimo", using non-toxic puffer elaborated by the techniques of netcage and land culture]. PMID- 15794097 TI - [Implementing HACCP system in small and/or less developed business--human development]. PMID- 15794098 TI - [Semicarbazide in food]. PMID- 15794099 TI - [Neuroprotective effects of novel derivatives of vasoactive intestinal peptide and pituitary adenylate cyclase-activating peptide in two brain ischemic models on mice]. AB - BACKGROUND: Vasoactive intestinal peptide (VIP), and pituitary adenylate cyclase activating peptide (PACAP), are members of a VIP/secretin/glucagon family. These peptides were demonstrated to possess the neuroprotective properties. However, these peptides are not suited to be developed as a medicine for brain ischemia because of their susceptibilities to endopeptidases. METHODS: We examined the effects of IK 312548 (IK), VIP derivative, and Ac-PACAP, PACAP derivative, on the 10 min two-vessel occlusion (2 VO) model in C 57 BL/6 N mice lacking a part of the posterior communicating artery, and the 30 min middle cerebral artery occlusion (MCAO) model in ICR mice. A 10 ml x kg(-1) dose of each derivative (final concentration; 1 fmol x kg(-1) and 100 pmol x kg(-1)) was injected intraperitoneally (i.p.) to each animal just after the preparation of brain ischemia. RESULTS: In 2 VO experiments, the number of neuronal cells in hippocampus was significantly reduced. However IK and Ac-PACAP treatments inhibited such reductions of neuronal cells in a dose-dependent manner. Particularly, between 1 pmol x kg(-1) and 100 pmol x kg(-) IK, and also between 10 fmol x kg(-1) and 1 pmol x kg(-1) Ac-PACAP significantly protected neuronal cell loss. In MCAO experiments, more than 60% of hemisphere was damaged. By treatment of IK (1-100 pmol x kg(-1)) and Ac-PACAP (1 fmol-1 pmol x kg(-1)), the range of brain damage decreased in a dose-dependent manner. CONCLUSIONS: Ac-PACAP and IK after the brain ischemia could pass the blood-brain barrier and protect brain cell damage. PMID- 15794100 TI - [The optimal dose of fentanyl added to 0.2% ropivacaine for postoperative epidural analgesia after gynecological surgery]. AB - BACKGROUND: Combined administration of local anesthetics and an opioid is frequently used in order to minimize the dose of each drug and to reduce adverse effects. Although fentanyl is commonly administered with local anesthetic, side effects of fentanyl increase in a dose-dependent manner. In this study, we determined the optimal dose of epidural fentanyl after gynecological surgery. METHODS: One hundred and sixteen adult patients scheduled for elective gynecological surgery were divided into 3 groups according to postoperative epidural analgesics; 0.2% ropivacaine (group R), 0.2% ropivacaine with 2 microg x ml(-1) fentanyl (group RF 2), or 0.2% ropivacaine with 5 microg x ml(-1) fentanyl (group RF 5). Each analgesic was infused at 5 ml x hr(-1) for 48 hr. Pain scores , incidence of NSAIDs administration and side effects were recorded for 48 hr after the surgery. RESULTS AND CONCLUSIONS: Ropivacaine alone could not provide sufficient analgesia. Although the addition of 5 microg x ml(-1) fentanyl to 0.2% ropivacaine at a rate of 5 ml x hr(-1) improved postoperative pain, side effects caused by fentanyl increased. Supplementing 2 microg x ml(-1) fentanyl provided sufficient analgesia with the least incidence of side effects. PMID- 15794101 TI - [Exhaled tidal volume is overestimated due to cardiogenic oscillation: effects of ventilator and mode]. AB - BACKGROUND: We investigated effects of cardiogenic oscillation on overestimation of tidal volume using a lung model, with three ventilators and two ventilatory modes. METHODS: We simulated cardiogenic oscillation at a rate of 90 breaths x min(-1) by ventilating one bellow of a two-bellow-type lung model. The magnitude of cardiogenic oscillation was defined as peak expiratory flow fluctuation when the airway was opened to the atmosphere. The lung model was mechanically ventilated with three ventilators (Bird 8400 STi, Servo-300, and Nellcor Puritan Bennett 840), two ventilatory modes (volume- and pressure-controlled ventilation), and two respiratory rates (5 and 10 breaths x min(-1)) in random order. We recorded tidal volume on a ventilator monitor and calculated the discrepancy from the set tidal volume. RESULTS: With Bird 8400 STi, monitored tidal volume exceeded set tidal volume, regardless of volume- or pressure controlled ventilation. The overestimation in tidal volume was larger with smaller respiratory rate and with larger cardiogenic oscillation. In contrast, with the other ventilators, the discrepancy was small. CONCLUSIONS: Exhaled tidal volume is overestimated during mechanical ventilation when cardiogenic oscillation is large. PMID- 15794102 TI - [Preoperative evaluation of hemodynamics using indocyanine green clearance meter in patients with peritonitis from gastrointestinal perforation]. AB - BACKGROUND: Since patients with perforated peritonitis often present hemodynamic instability, preoperative evaluation of hemodynamics is important for anesthesiologists. Indocyanine green (ICG) can be used to measure cardiac output (C.O.), blood volume (B.V.) and hepatic function. Previous studies have reported that the average elimination rate constant of ICG (KICG) might be reduced in critically ill patients. This study was therefore conducted to investigate the relationship between acute physiology and chronic health evaluation (APACHE) II score and KICG value, cardiac output, and blood volume in patients scheduled for emergency surgery. METHODS: After obtaining informed consent hemodynamics of thirteen patients undergoing emergency surgery for perforated peritonitis were measured by in vivo ICG clearance technique. After induction of anesthesia, ICG elimination rate was measured following intravenous administration of ICG (20 mg) with a DDG analyzer (DDG-2001) using a nose probe for pulse dye densitometry. All measurements were performed prior to surgery. The value of C.O./B.V. was used as an indicator of cardiac function. The severity of each patient's illness was also assessed using the APACHE II scoring system at the start of operation. RESULTS: There was a negative correlation between APACHE II score and KICG (KICG = 0.34 0.01 x APACHE II score, r = -0.56). In addition, there was a positive correlation between APACHE II score and C.O./B.V. (C.O./B.V. = 1.10 + 0.05 x APACHE II score, r = 0.79). CONCLUSIONS: Patients with high APACHE II score may show low hepatic blood flow and high cardiac output compared with blood volume. ICG clearance rate might be useful for preoperative evaluation of hemodynamics in patients with perforated peritonitis. PMID- 15794103 TI - [Evaluation of 38 cases of employing aortic occlusion balloon catheter]. AB - BACKGROUND: Aortic occlusion balloon catheter (AOBC), which occludes the descending aorta without thoracotomy, is expected to prevent further blood loss and raise blood pressure (BP). We investigated the effects of AOBC retrospectively. METHOD: AOBC was used in 38 patients for perioperative management only if the BP responded insufficiently despite rapid resuscitation due to massive hemorrhage below the diaphragm. There were 33 trauma cases, and five cases of ruptured abdominal aortic aneurysm. We inserted AOBC via the femoral artery or left common carotid artery in the emergency room(ER) or in the operating room (OR). RESULTS: BP increased for 38.7 +/- 33.9 mmHg following AOBC, with a survival rate of 36%. CONCLUSIONS: In our experience, AOBC was effective for raising BP in patients in hemorrhagic shock. PMID- 15794104 TI - [Anesthetic management for video-assisted extended thymectomy of patients with myasthenia gravis]. AB - BACKGROUND: Thymectomy is one of therapeutic modalities for patients with myasthenia gravis. Since 1998, we have performed video-assisted extended thymectomy, which is much less invasive than traditional method. However, its optimal perioperative management has not been established. METHODS: From April 1998 to December 2003, 40 patients with myasthenia gravis underwent video assisted extended thymectomy in our hospital. Anesthesia was maintained with propofol, sevoflurane and epidural anesthesia. No muscle relaxant was used. Operations were performed in supine position and required differential lung ventilation of both sides in turn for manipulation. A central venous catheter was inserted in the femoral vein to prepare for unexpected bleeding or other hemodynamic changes. RESULTS: Seven patients presented hypoxemia under differential lung ventilation and needed bilateral lung ventilation or addition of CPAP to nondependent lung. Unexpected bleeding from the left innominate vein occurred in two patients and required median sternotomy. Severe hypotension caused by compression of the heart during operation was observed in twenty-one patients. Extubation in the operating room was successful in all patients except one with severe bulbar paralysis before preoperative period. CONCLUSIONS: It is important to examine both surgical techniques of video-assisted surgery and physiological features of myasthenia gravis for anesthetic management of video assisted extended thymectomy. PMID- 15794105 TI - [Anesthetic management in ten cases for tracheobronchial Dumon stent placement]. AB - BACKGROUND: To determine perioperative complications, we evaluate herein 10 cases of anesthetic management for placement of Dumon stent in patient with tracheal or bronchial stenosis due to invasion of esophageal or lung cancer. METHODS: After sufficient oxygenation, anesthesia was induced with propofol and fentanyl. Since muscle relaxant has been considered safe for central-type air way stenosis except for cases involving large anterior mediastinal masses, we administered vecuronium for all cases to facilitate insertion of rigid bronchoscope and for surgical procedures. Anesthesia was maintained with continuous infusion of propofol, and ventilation was performed via a side-port of a rigid bronchoscope with 100% oxygen. Extra corporeal circulation was instituted in 2 cases. RESULTS: In 5 of the 10 cases, stent placement was uneventful. However, in the other 5 cases, respiratory failure (SpO2 < 90% and/or PaCO2 > 80 mmHg: 4 cases) or severe hypotension (systolic blood pressure < 60 mmHg: 3 cases) developed. Severe hypotension was attributed to relatively higher dose of anesthetic agents for cachexic status, or reduction in venous return following over-inflation of the lungs. Acute reduction in blood carbon dioxide levels due to extracorporeal circulation (case 4), and loss of consciousness after administration of anesthetic agents (case 2) could also have been involved in 2 cases. CONCLUSIONS: Circulatory status must be closely monitored during anesthetic management for Dumon stent placement. PMID- 15794106 TI - [Nitroglycerin to relax the uterus during cesarean delivery for the low birthweight baby]. AB - BACKGROUND: The uterus at premature birth being thick and sensitive, we applied nitroglycerin (NTG) to relax the uterus during cesarean delivery for the low birthweight baby. METHODS: Twelve parturients of impending premature birth and nine of severe intoxication were included. General anesthesia was induced with thiamylal, and maintained with 5% inhaled sevoflurane. NTG was injected in an intermittent dose of 500 microg. The relative hardness of the uterus at delivery was evaluated by obstetricians in four ranks of categories. RESULTS: The mean gestation period was 27.4 weeks (24 weeks at the minimum-33 weeks at the maximum) and the mean birthweight was 986 g (561-1518 g). The mean time elapsed from induction of anesthesia to delivery was 8.3 min (2-14 min) while NTG was injected for the mean 1.8 times that reflected a total dose of 900 microg (500-1500 microg). The mean endtidal concentration of sevoflurane at delivery reached 3.8% (3.4-4.3%). The uterus was evaluated as "very soft" in 15 out of 21 cases, and all babies were extracted easily without injury through the lower J-shaped section of the uterus. The mean Apgar score was 2.9 points at 1 minute and 6.3 points at 5 minutes. No adverse effect of NTG on the mothers or babies was noticed. CONCLUSIONS: NTG relaxes the uterus effectively and facilitates cesarean delivery for the low birthweight baby. PMID- 15794107 TI - [A case of thoraco-amniotic shunt for congenital cystic adenomatoid malformation]. AB - Congenital cystic adenomatoid malformation (CCAM) is a congenital pulmonary anomaly, which may lead to fetal hydrops, pulmonary hypoplasia, and fetal or neonatal death. Recently, diagnosis and surgery for fetus have been improved. We experienced a case of CCAM, classified in Stocker class I, with a single cyst of about 5 cm in diameter. A 32 year-old pregnant woman had a fetus diagnosed as CCAM at 25 th gestational week. The thoraco-amniotic shunt placement using a modified double pig tail catheter was performed at 27 th gestational week under general anesthesia with 1% isoflurane in oxygen 2 l x min(-1) and nitrous oxide 4 l x min(-1), and 100 microg of fentanyl. Fetal movement was suppressed during surgery. This fetal therapy was effective and the cyst disappeared after surgery. The baby was delivered with caesarian section under spinal anesthesia with 0.5% bupivacaine 2.5 ml. On the next day, right lung lobectomy was accomplished under general anesthesia with fentanyl 0.2 mg and pancuronium 6.0 mg. Anesthesia with isoflurane, nitrous oxide, and fentanyl was useful for the fetal surgery of thoraco-amniotic shunt placement. Consequently, caesarian section and lung lobectomy were completed successfully. PMID- 15794108 TI - [Retrograde nasal intubation for a patient with difficult airway by using 28 Fr double lumen tube]. AB - We experienced retrograde nasal intubation with a 28 Fr double lumen tube (DLT) in a patient with difficult airway. A 56-year-old man who had undergone an operation for carcinoma of the floor of the mouth was scheduled for right upper lobectomy with video-assisted thoracic surgery. Because of the postoperative anatomical anomaly, the usual laryngoscopy and retrograde oral intubation was not successful. So we reduced the tube size to 28 Fr and succeeded in retrograde nasal intubation. The positioning of the DLT was confirmed by auscultation and clinical signs. We could complete one lung ventilation. There was no trouble of inflation and deflation in anesthesia term. We conclude that 28 Fr DLT is one of the useful tools for difficult airway. PMID- 15794109 TI - [Anesthetic management of an 18-trisomy neonate with high frequency ventilation]. AB - Neonates with 18-trisomy syndrome have various anomalies including cardiac and facial anomalies. Active treatments for them have not been recommended due to a low survival over one year which is less than 10%. However, the survival over one year in our institute between 1985 and 2003 is higher at 22%, and we had a patient with survival of over 5 years. We report a case of anesthetic management in a male 18-trisomy neonate who received a radical repair of umbilical hernia immediately after his birth. Tracheal intubation was difficult due to small airway and facial anomalies; cleft clip, cleft palate, and macrognathia. Conventional ventilation was ineffective for elimination of carbon dioxide and oxygenation due to pulmonary hypoplasia and intra- and extra-cardiac shunts. A high frequency ventilation improved elimination of carbon dioxide and oxygenation. PMID- 15794110 TI - [Overdose of vecuronium during general anesthesia to an infant]. AB - We experienced an accidental case in which 5 mg of vecuronium was administered to a 5 kg infant boy, aged 2 months, incrementally for 55 minutes during general anesthesia. He received general anesthesia at the hospital where there is no anesthesiologist. After surgery the surgeon felt an unusual situation during emergence period from general anesthesia and then noticed overdose of vecuronium. They got in touch with our department immediately and requested us to follow him up. When we reached at the bedside, we could not observe voluntary movement by him. By using neuromuscular monitoring, we could determine that a paralysis was still maintained. After a while, he started to move and breathe himself. Extubation was performed safely 4 and half hours after the final administration of vecuronium. Finally, he could recover with no adverse effect. To predict the optimal dose of a non-depolarizing muscle relaxant to a pediatric patient, routine monitoring of neuromuscular block is effective and essential. PMID- 15794111 TI - [Anesthesia for cesarean section in a parturient taking risperidone and haloperidol]. AB - We report a case of anesthesia for cesarean section in a schizophrenic patient. Her psychiatric symptoms were well controlled with low doses of risperidone until 35 weeks' gestation, when she suddenly developed psychotic manifestations. Risperidone 6 mg x day(-1) and haloperidol 12 mg x day(-1) PO were given for 3 weeks before delivery. Elective cesarean section was performed under spinal anesthesia at 38 weeks. The parturient showed good psychiatric condition during and after the surgery. The neonate did not show any symptoms which antipsychotics could have caused. Maternal and umbilical blood concentrations of risperidone and haloperidol are reported. PMID- 15794112 TI - [Anesthetic management of an infant with asplenia and single atrium single ventricle undergoing ear tube surgery for otitis media: a case report]. AB - A 13-month-old girl with cyanotic congenital heart disease; single atrium, single ventricle, common atrioventricular (AV) valve, pulmonary atresia and total anomalous pulmonary venous drainage, suspected of asplenia underwent ear tube surgery for otitis media. She had undergone bilateral Blalock-Taussig shunts for her heart disease. She had congestive heart failure due to moderate to severe common AV valve regurgitation and often experienced respiratory tract infection with sputum. Oxyhemoglobin saturation measured by pulse oximetry was 75-80% and polycythemia was found in complete blood count. We chose tracheal intubation for her airway management because of a large amount of sputum. General anesthesia was maintained with sevoflurane, nitrous oxide and oxygen for ear tube surgery. During anesthesia she showed several episodes of desaturation which were well managed by frequent tracheal suctioning. Her circulation was stable with 50% N2O and sevoflurane 1.7-2.0%. The operation was performed uneventfully and the patient was discharged to the ward after tracheal extubation. Asplenia is frequently complicated with cyanotic congenital heart disease and increased susceptibility to bacterial infection. Anesthesia for these patients with upper respiratory infection should be managed with tracheal intubation even for a short surgery. PMID- 15794113 TI - [Which is more useful for central venous catheterization? A comparison between a 22 gauge metal needle (Argyle) and a 24 gauge plastic cannula with a metal needle (Insyte)]. AB - BACKGROUND: Central venous catheterization is commonly performed by puncturing the internal jugular vein. However, placement of central venous catheters is not without risk. We compared the Argyle and the Insyte in terms of usefulness and incidence of complications during central venous catheterization. METHODS: Twenty adult patients for cardiac operation were randomly allocated into two groups; 10 patients in the Argyle group (Group A) and 10 patients in the Insyte group (Group B). RESULTS: In both groups, no kinking or stucking of the guide wire occurred during insertion. No accidental puncture of the carotid artery occurred in Group B, while it occurred in one case in Group A. The time required for insertion of the guide wire was not significantly different between the two groups, though it tended to be shorter in Group B than in Group A (19.3 +/- 14.6 sec vs 34.6 +/- 34.0 sec). On ultrasonography, the Argyle was observed to compress the internal jugular vein and to be prone to penetrate both anterior and posterior walls. CONCLUSIONS: Using the Argyle makes it a little more difficult to place the needle in the appropriate venous lumen. We conclude that central venous catheterization with the Insyte was quicker and safer than with the Argyle. PMID- 15794114 TI - [Quantitative assessment of pressure relief at the sacral area in adults lying supine on the operating room table]. AB - BACKGROUND: It is important to prevent development of the pressure ulcers in patients undergoing lengthy surgery, particularly at areas of skin overlying bony prominences. This study was designed to investigate distribution of the interface pressure (IP) over the body area (from the head to pelvic area) in supine adults and also evaluate the ability of a polyurethane-made cushion to reduce the IP at their sacral area. METHODS: Utilizing a recently developed device to measure the IP (ERGO-CHECK, ABW Co., Germany), we evaluated distribution of the IP (estimated per 3 x 4 cm2 area) over the body area in healthy volunteers (n=31) and patients under general anesthesia (n=6) lying supine on the operating room (OR) table. RESULTS: In all the subjects, the highest IP was generated at the sacrum; 62.5 +/ 23.8 (mean +/- SD) and 35.7 +/- 5.5 mmHg in the volunteers and patients, respectively. The polyurethane-made, "doughnut" cushion (5 cm in thickness) inserted between the pelvic area and the OR table significantly reduced (P < 0.05) the IP at the sacrum in both groups: the IPs after the insertion in the volunteers and patients were 35.1 +/- 11.1 and 25.6 +/- 6.5 mmHg, respectively. In addition, the insertion significantly reduced (P < 0.05) the high-risk area (i.e., area of IP > 32 mmHg) in both groups. CONCLUSIONS: Quantitative assessment of the IP would be useful in evaluating precisely the effectiveness of various types of pillows, cushions, or mattresses designed to reduce the IP. PMID- 15794115 TI - [Complications related to anesthesia method in the University of Occupational and Environmental Health Hospital]. AB - BACKGROUND: Complications related to anesthesia remain a problem. We studied the incidence of complications during anesthesia in 2758 patients who had undergone anesthesia in the University of Occupational and Environmental Health Hospital. METHODS: We checked the anesthesia records retrospectively and analyzed the collected data for the incidence of complications during anesthesia. RESULTS: The total incidence of complications during anesthesia was 12.2%. The incidences of complication are estimated to be 13.4% in inhalation anesthesia, 11.9% in inhalation anesthesia plus epidural, spinal or conduction block, 8.9% in CSEA, zero % in epidural anesthesia and 7.5% in spinal anesthesia. CONCLUSIONS: The incidence of complications in inhalation anesthesia was almost as same as that in inhalation anesthesia plus epidural, spinal or conduction block. More study should be necessary to prevent complications related to anesthesia. PMID- 15794116 TI - [Development of [18F]FRP-170 injection for imaging hypoxia by PET]. AB - A novel [18F]FRP-170 injection for imaging hypoxia by PET was developed for clinical use. The preparation was based on the simple on-column basic-hydrolysis and the whole procedure was automated by detecting He flow change for transferring and evaporating liquids. [18F]FRP-170 was prepared in around 15-20% decay-corrected radiochemical yield within 60 min and stable in saline for more than 6 hr. Radiochemical purity was over 99% and specific activity at EOS was 40 60 GBq/micromol. The radiation-absorbed dose to the whole body was estimated to be 1.0 mSv/185 MBq. The [18F]FRP-170 injection proved to be suitable for clinical use without acute toxicity or mutagenicity. PMID- 15794117 TI - [Improvement in accuracy of quantitative assessment of the regional cerebral blood flow with 99mTc-ECD]. AB - Mean cerebral blood flow (mCBF) in the slice including the basal ganglia (reference slice) is necessary for the quantification of regional CBF using Patlak plot and BUR methods on 99mTc-ECD cerebral perfusion SPECT. The mCBF was calculated from the mean counts of this slice. A region of interest (ROI) has been manually set on the reference slice to obtain the mean counts (manual ROI method). However, there was large variability observed in the value of rCBF in this method. We developed a 3DSRT method for improving the accuracy of the mean counts in the reference slice and evaluated the difference between the value of rCBF on manual ROI method and that on 3DSRT method in consecutive 11 patients with cerebral vascular disease. Difference in the value of mean counts of the reference slice was distributed within the 2 standard deviations (SD) with Blant Altman analysis in 9 of 11 patients. Significant difference in the value of mean counts between two methods was observed in 2 of 11 patients. 3DSRT method is superior accuracy to the manual ROI method in the evaluation of the counts in the ROI. Lower accuracy in manual ROI method, therefore, results in the difference of the value of mean counts. 3DSRT method provides high accuracy with the various quantitative methods for the evaluation of rCBF using 99mTc-ECD. PMID- 15794118 TI - [Guidelines for radio-iodine (131I) therapy in Graves' disease and thyroid cancer]. AB - Radio-iodine (131I) therapy has been using in Graves' disease and well differentiated thyroid cancer. The rules of control in the discharge from radio isotope hospital were notified in 1999 in Japan. Guideline of the 131I therapy in Graves' disease and thyroid cancer were prepared by sub-group of Japanese Society of Nuclear Medicine. PMID- 15794119 TI - A sensitive and rapid chemiluminescence ELISA for filamentous bacteriophages. AB - Filamentous bacteriophage (Ff) displayed random peptide and antibody libraries are widely used to identify specific, high affinity, binding ligands. A critical element in the identification of target-specific phages is to determine phage titers, not only at every round of selection, but also for normalizing phage titers of a set of individual clones for their comparative binding analysis. The conventional ELISA-based Ff titration methods require a minimum of 4-5 hr assay time and their lowest reported detection limit is approximately 50,000 particles/well. In this report, we present a sandwich ELISA that allows detection of approximately 1000 Ff particles/well in less than 2.5 hr. The values of correlation of coefficient (r2) for the curves at low phage concentrations (up to 106 TU/well) were about 0.999 in our ELISA. Experiments conducted at different temperatures suggest using 40 degrees C incubations when titering low phage concentration samples. Experiments were also conducted with conventional ELISA for comparison. Our ELISA method derives an advantage from using a chemiluminescence substrate that gives much larger signals and wide linear range of measurement, thus allowing discrimination between background and low Ff phage concentrations. In conclusion, the Ff titration method presented here is highly sensitive, rapid, and amenable to high throughput analysis. PMID- 15794120 TI - Kinetics of androstendione-radioactive immunocomplex substitution reaction. AB - A kinetic model is put forward for the study of the antigen-antibody reactions involved in the coated tube radioimmunoassay (RIA) of androstendione. Twenty experiments were conducted to determine the influence of initial concentrations, ionic strength, viscosity, and temperature on the substitution reaction of 125I androstendione (M) by unlabelled androstendione (Q) in the immunocomplex PM (P = anti-androstendione antibody). The results obtained are in line with the proposed model. The concentration of radioactive immunocomplex is directly proportional to the initial concentration of labelled androstendione and independent of the concentration of unlabelled androstendione, ionic strength, and viscosity. The reaction is not diffusion-controlled. PMID- 15794121 TI - A new efficient method for eliminating the interference effect of human serum and increasing the sensitivity and recovery rate of enzyme immunoassay. AB - A new, very simple method for increasing the sensitivity and recovery rate of enzyme-linked immunosorbent assay (ELISA) for the precise quantification of antigen in human serum is described. The assay design uses CATNF6A4c IgG2a monoclonal antibody and biotinylated anti-human tumor necrosis factor-alpha (hTNF alpha) polyclonal mouse IgG as the capture and tracer antibodies, respectively. The assay is completed within 4 hours at room temperature and is capable of detecting both recombinant and native human TNF-alpha. The assay incorporates the use of saturated ammonium sulfate (SAS) as a component of the dilution buffer to amplify the resultant signal from antigen containing human serum and eliminating the endogenous interference of native human serum. SAS worked optimally at the final concentrations, ranging from 1.2% to 11%. The addition of SAS to the dilution buffer resulted in a dramatic increase in both sensitivity and recovery rate of the ELISA. The results demonstrated that 50 microL of dilution buffer, containing SAS, enabled the precise quantification of human TNF-alpha in 100 microL of human serum samples and eliminated the interference of native serum, which seemed to be related to complement proteins. Therefore, dilution buffer containing SAS, at a defined concentration, seemed to be a potential candidate for resolving sensitivity and recovery problems usually encountered in immunoassays when measurement was performed with native serum samples. The proposed technique provides an easy, practical, and consistent method for ELISA when using human native serum. PMID- 15794122 TI - Quantitative chemiluminescent immunoassay for NF-kappaB-DNA binding activity. AB - Nuclear factor-kappaB (NF-kappaB) is a ubiquitous redox-sensitive transcription factor involved in the pro-inflammatory response to several factors, including cytokines and oxidative stress. Upon activation, NF-kappaB translocates into the nucleus and binds to specific nucleotide sequences. The cellular responses to inflammatory and stress signals have been implicated in disease conditions, such as atherosclerosis, cancer, diabetes, and Alzheimer's disease. The conventional method for detection of NF-kappaB -DNA binding activity is the electrophoretic mobility shift assay (EMSA), which is time-consuming and non-quantitative. Here, we report (a) development of a rapid, sensitive and quantitative chemiluminescent immunoassay (QCI) for analysis of NF-kappaB DNA-binding activity, and (b) validation of the QCI with the EMSA using nuclear and cytosolic extracts from cultured prostate cancer cells (PC3), rat liver homogenates and human lymphocytes. The QCI for analysis of NF-kappaB DNA binding activity has advantages over the EMSA: (1) Higher speed: 3-5h post sample preparation, (2) Greater sensitivity: 10pg NF-kappaB/well, (3) Quantitative: linear range: 10 1000pg NF-kappaB; r2 = 0.999 (4) High throughput adaptability: 96-well plate format can analyze up to 40 samples in duplicate, (5) SAFETY: No radioactive isotopes, (6) Simplicity, and (7) Capability of measurement of both activated (free) NF-KB which is translocated into the nucleus and total (bound + unbound) NF-kappaB present in the cytosol/cell. PMID- 15794123 TI - The isolation and characterization of human natural alphaGal-specific IgG antibodies applicable to the detection of alphaGal-glycosphingolipids. AB - The Galalpha1-3Galbeta (alphaGal) hapten is xenogeneic for humans; natural anti alphaGal antibodies are present in human serum. To study the possible abnormal expression of the alphaGal in humans and the pathophysiological role of antibodies, the method of affinity purification of human anti-alphaGal IgG was developed. The specificity of antibodies was evaluated using polyacrylamide (PAA) based glycoconjugates in direct and competitive enzyme-linked immunosorbent assays (ELISA). The purified antibodies exhibited alphaGal-restricted specificity. The IC50 value for alphaGal-PAA was equal to 4 x 10(-8) M. In a competitive assay, the Galalpha1-3(Fucalpha1-2)Galbeta-PAA (trisaccharide of blood group B) was found to be one hundred times less active inhibitor than alphaGal-PAA. The multivalent alphaGal-PAA was 1100 times more potent an inhibitor than the monovalent spacered alphaGal-saccharide. The antibodies did not show any reactivity to the negatively charged antigens (DNA, human tumor derived mucins). At a concentration of 2 microg/mL, the antibodies agglutinated rabbit erythrocytes but not hare erythrocytes. The high reactivity of antibodies to the alphaGal-glycosphingolipids of rabbit erythrocytes and the pig kidney was shown by a modified sensitive method of thin-layer chromatography with immunodetection. PMID- 15794124 TI - Development and validation of a simple, sensitive, second antibody format enzyme immunoassay (EIA) for LH determination in mithun (Bos frontalis) plasma. AB - The objective of this study was to develop and validate a simple and highly sensitive enzyme immunoassay (EIA) for LH determination in mithun plasma on microtitreplates using the biotin-streptavidin amplification system and the second antibody coating technique. Biotin was coupled to LH and used to bridge between streptavidin-peroxidase and immobilized antiserum in competitive assay. The EIA was carried out directly in 20 microL mithun plasma. The LH standards ranging from 6.25 pg/well/20 microL to 400 pg/well/20 microL were prepared in hormone free plasma collected from a mithun on day 3 post calving. The sensitivity of EIA procedure was 6.25 pg/well LH, which corresponds to 0.31 ng/mL plasma; the 50 percent relative binding sensitivity was seen at 100 pg/well/20 microL. Plasma volumes for the EIA viz. 10 and 20 microL did not influence the shape of standard curve even though a slight drop in the OD450 was seen with higher plasma volumes. A parallelism test was carried out to compare the endogenous mithun plasma LH with bovine LH standards. It showed good parallelism with the bovine standard curve. For the biological validation of the assay, 3 mithuns were used. These were administered 10 microg i.v., with a synthetic analogue of GnRH (Buserelin-Acetate, Intervet, India) and blood samples were collected at 15 min intervals using indwelling jugular catheter beginning 1 h prior to GnRH injection till 8 h post injection. In all animals, sharp increases in LH concentrations were recorded post GnRH administration, which confirms the biological validation of the EIA. In conclusion, the EIA developed for LH determination in mithun blood plasma is sufficiently reliable, economical, and sensitive enough to estimate LH in all physiological variations in mithun. PMID- 15794125 TI - Topography and function of androgen-metabolizing enzymes in the central nervous system. AB - The present review describes concisely the topography and function of the three androgen-metabolizing enzymes, namely aromatase, 5alpha-reductase and 3alpha hydroxysteroid dehydrogenase, in the central nervous system (CNS). Aromatase, estrogen synthetase, is the key enzyme for converting androgens to estrogens. Aromatase is indispensable for the sexual differentiation of the brain and the enzyme activity and expression of aromatase are high during the critical period of neural development, which extends from the late embryonal to the early neonatal period in rodents. Aromatase is expressed in neurons within specific hypothalamic and limbic regions. The locations of aromatase-immunoreactive neurons are divided into three groups according to the period of enzyme expression. Steroid 5alpha-reductase converts a number of steroids with a C3 ketone group and a C4-C5 double bond (delta4; androgens, progestins and glucocorticoids) to their 5alpha-reduced metabolites. Two isoforms of 5alpha reductase are found and type 1 is predominant in neural tissues. The enzyme activity of 5alpha-reductase is found widely in the CNS and is high in white matter regions. The enzyme expression of 5alpha-reductase peaks during the late embryonic period. 3alpha-Hydroxysteroid dehydrogenase is the oxidoreductase that interconverts 3-ketosteroids to 3alpha-hydroxysteroids. Four isozymes have been found in humans and only one type has been found in rats. The enzyme converts 5alpha-reduced steroids (e.g. 5alpha-dihydroprogesterone) to tetrahydrosteroids (e.g. 3alpha,5alpha-tetrahydroprogesterone). The latter steroid is a potent stimulator of the GABA(A) receptor. The activity of 3alpha-hydroxysteroid dehydrogenase is high during the first 1-2 postnatal weeks, decreases with development and this enzyme is highly expressed in astrocytes. PMID- 15794126 TI - Cell fate determination regulated by a transcriptional signal network in the developing mouse brain. AB - Development of the nervous system is governed by multistep processes in which neurons, astrocytes and oligodendrocytes are produced from common neural stem cells. Differentiation of stem cells into these three types of neural cells is directed, in part, by cell-external cues, such as cytokines. We have examined the effects of astrocyte-inducing cytokines, leukemia inhibitory factor (LIF) and bone morphogenetic protein (BMP) 2 on mouse telencephalic neuroepithelial cells. We found that simultaneous treatment of neuroepithelial cells with LIF and BMP2 synergistically induced astrocytic differentiation. The synergistic effect of the two cytokines is brought about by a complex formation composed of downstream transcription factors signal transducer and activator of transcription (STAT) 3 and Smad1, together with the transcriptional coactivator p300. Recent studies also clarified molecular interactions involving basic helix-loop-helix-type transcription factors that promote differentiation of neuronal or oligodendrocytic cell lineages. For instance, in fetal brain, it has been shown that the neurogenic transcription factor Neurogenin1 inhibits formation of a STAT3/p300/ Smad1 complex and that the oligodendrocytic transcription factor OLIG2 does the same. This suggests the presence of mutual negative interaction among the signaling pathways regulating the three neural cell lineages. Furthermore, we have demonstrated the developmental stage-dependent epigenetic regulation of cell type-specific gene promoters. DNA methylation, histone acetylation and modified-DNA binding proteins are thought to be involved in this process. In the present review, we discuss the cross-talk between transcription factors downstream of the cytokine-signaling pathways and their effects on cell fate determination. We also emphasize the importance of epigenetic modification of lineage-specific gene promoters and chromatin structure in transcription factor-mediated cell fate regulation. PMID- 15794127 TI - Involvement of filamin A and filamin A-interacting protein (FILIP) in controlling the start and cell shape of radially migrating cortical neurons. AB - Precisely regulated radial cell migration out of the ventricular zone is essential for corticogenesis. However, molecular mechanisms controlling the start of migration and the dynamics of migrating cell shape remain elusive. Here, we show novel mechanisms that can tether ventricular zone cells and control migrating cell shape. The novel protein Filamin A-interacting protein (FILIP) interacts with Filamin A, an indispensable actin-binding protein for cell motility, and induces its degradation in COS-7 cells. Degradation of Filamin A is indicated in the cortical ventricular zone where FILIP mRNA localizes. Furthermore, most ventricular zone cells that overexpress FILIP fail to migrate in explants. These results indicate that FILIP acts through a Filamin A-F-actin axis to control the start of neocortical cell migration from the ventricular zone. Filamin A also determines the shape of migrating neocortical neurons, which show global morphological changes and complicated behavior during that migration. Dysfunction of Filamin A, caused by a mutant Filamin A expression, prevents cells from acquiring consistent polarity toward specific direction and decreases motility in the subventricular and intermediate zones. In contrast, Filamin A overexpression, achieved by a short interfering RNA for FILIP, promotes the development and maintenance of a bipolar shape also in the subventricular and intermediate zones. These results suggest that the amount of Filamin A helps migrating neurons determine their mode of migration, multipolar or bipolar, prior to entering the cortical plate and that FILIP is responsible, at least in part, for the Filamin A content of migrating neurons. PMID- 15794129 TI - Diverse functions of the p75 neurotrophin receptor. AB - The pan-neurotrophin receptor p75NTR belongs to a large family of receptors, which includes tumor necrosis factor receptors, Fas and approximately 25 other members. The p75NTR is the first receptor to be cloned molecularly. Recent years have seen the emergence of a consensus regarding the signaling pathways activated by p75NTR and its potential biological function, although receptor characterization had not been targeted for some years. We now know that p75NTR has surprisingly diverse effects, ranging from cell death to regulation of axon elongation. This diversity can be explained by the complex formation of p75NTR with other receptors and multiple signaling molecules that interact with the intracellular domain of p75NTR. PMID- 15794128 TI - Signaling cascade coordinating growth of dorsal and ventral tissues of the vertebrate brain, with special reference to the involvement of Sonic Hedgehog signaling. AB - The vertebrate brain is a complex and highly organized structure with numerous neurons and glial cells. During development, undifferentiated progenitor cells proliferate from neural stem/precursor cells and gradually restrict their fates according to their environment. Differentiated cells are arranged precisely to accomplish their function and to maintain integrity as a whole brain. In this respect, cells must receive signals to know where and when they determine their fates. Secreted and membrane molecules convey the information between cells. The secreted glycoprotein Sonic hedgehog (Shh) is one of such signaling molecules. Sonic hedgehog is widely known to specify ventral neuronal types according to the concentration of Shh, whereas differentiation of dorsal neurons is largely independent of Shh. However, in the diencephalon and midbrain, dorsal parts are also affected in Shh-mutant embryos. Detailed analysis demonstrated that Shh signaling indirectly regulates the growth of the dorsal tissue in these regions. One of the fibroblast growth factor (FGF) members, namely FGF15, has been reported to be downstream to Shh signaling in the mouse embryonic brain. Luciferase assays and transgenic analysis revealed that the Fgf15 gene is a direct target of Shh. Downregulation of Tcf4 and upregulation of Bmp4 in Shh mutants suggest that Wnt and BMP signals from the dorsal midline are also involved in the dorsal brain phenotype. These data suggest the coordinating role of the Shh-FGF15-Wnt/BMP signaling cascade between the ventral and dorsal parts of the brain. PMID- 15794130 TI - Regulation of neural development by glial cell line-derived neurotrophic factor family ligands. AB - Glial cell line-derived neurotrophic factor (GDNF) and its three relatives constitute a novel family of neurotrophic factors, the GDNF family ligands. These factors signal through a multicomponent receptor complex comprising a glycosylphosphatidylinositol-anchored cell surface molecule (GDNF family receptor (GFR) alpha) and RET tyrosine kinase, triggering the activation of multiple signaling pathways in responsive cells. Recent gene-targeting studies have demonstrated that GDNF family ligands are essential for the development of a diverse set of neuronal populations and we have now started to understand how these ligands uniquely regulate the formation and sculpting of the nervous system. Recent studies have also revealed interactions by multiple extracellular signals during neural development. The deciphering of GDNF family ligand signaling in neural cells promises to provide vital new insights into the development and pathology of the nervous system. PMID- 15794131 TI - Oncostatin M in the development of the nervous system. AB - Oncostatin M (OSM) is a member of the interleukin-6 family of cytokines. Of these cytokines, OSM is closely related structually, genetically and functionally to leukemia inhibitory factor. However, OSM-specific biological activities have been reported in hematopoiesis and liver development. Recently, we have demonstrated OSM-specific activities in the nervous systems. In the adult central nervous system (CNS), OSM receptor (OSMR) beta was observed in meningeal cells of pia mater, epithelial cells of the choroid plexus and olfactory astrocyte-like glia surrounding the glomeruli of the olfactory bulb. In the CNS of neonatal mice, OSMRbeta was also expressed in the ventral subnucleus of the hypoglossal nucleus, but disappeared at post-natal day (P) 14. In contrast with the CNS, OSMRbeta was strongly expressed in small-sized non-peptidergic neurons of the dorsal root ganglia (DRG) and trigeminal ganglia (TG). Interestingly, all OSMRbeta-positive neurons in these ganglia also expressed both TRPV1 (a vanilloid receptor) and P2X3 (a purinergic receptor). In OSM-deficient mice, TRPV1/P2X3/OSMRbeta triple positive neurons were significantly decreased. Consistent with such histological findings, OSM-deficient mice exhibited a reduction in responses to various stimuli, including mechanical and thermal stimuli. These findings suggest an important role for OSM in the development of a subset of nociceptive neurons. PMID- 15794132 TI - Applications of Forman's discrete Morse theory to topology visualization and mesh compression. AB - Morse theory is a powerful tool for investigating the topology of smooth manifolds. It has been widely used by the computational topology, computer graphics, and geometric modeling communities to devise topology-based algorithms and data structures. Forman introduced a discrete version of this theory which is purely combinatorial. This work aims to build, visualize, and apply the basic elements of Forman's discrete Morse theory. It intends to use some of those concepts to visually study the topology of an object. As a basis, an algorithmic construction of optimal Forman's discrete gradient vector fields is provided. This construction is then used to topologically analyze mesh compression schemes, such as Edgebreaker and Grow&Fold. In particular, this paper proves that the complexity class of the strategy optimization of Grow&Fold is MAX-SNP hard. PMID- 15794133 TI - Visualizing a sphere eversion. AB - The mathematical process of everting a sphere (turning it inside-out allowing self-intersections) is a grand challenge for visualization because of the complicated, ever-changing internal structure. We have computed an optimal minimax eversion, requiring the least bending energy. Here, we discuss techniques we used to help visualize this eversion for visitors to virtual environments and viewers of our video The Optiverse. PMID- 15794134 TI - Robust feature detection and local classification for surfaces based on moment analysis. AB - The stable local classification of discrete surfaces with respect to features such as edges and corners or concave and convex regions, respectively, is as quite difficult as well as indispensable for many surface processing applications. Usually, the feature detection is done via a local curvature analysis. If concerned with large triangular and irregular grids, e.g., generated via a marching cube algorithm, the detectors are tedious to treat and a robust classification is hard to achieve. Here, a local classification method on surfaces is presented which avoids the evaluation of discretized curvature quantities. Moreover, it provides an indicator for smoothness of a given discrete surface and comes together with a built-in multiscale. The proposed classification tool is based on local zero and first moments on the discrete surface. The corresponding integral quantities are stable to compute and they give less noisy results compared to discrete curvature quantities. The stencil width for the integration of the moments turns out to be the scale parameter. Prospective surface processing applications are the segmentation on surfaces, surface comparison, and matching and surface modeling. Here, a method for feature preserving fairing of surfaces is discussed to underline the applicability of the presented approach. PMID- 15794135 TI - Fast evolution of image manifolds and application to filtering and segmentation in 3D medical images. AB - In many instances, numerical integration of space-scale PDEs is the most time consuming operation of image processing. This is because the scale step is limited by conditional stability of explicit schemes. In this work, we introduce the unconditionally stable semi-implicit linearized difference scheme that is fashioned after additive operator split (AOS) [1], [2] for Beltrami and the subjective surface computation. The Beltrami flow [3], [4], [5] is one of the most effective denoising algorithms in image processing. For gray-level images, we show that the flow equation can be arranged in an advection-diffusion form, revealing the edge-enhancing properties of this flow. This also suggests the application of AOS method for faster convergence. The subjective surface [6] deals with constructing a perceptually meaningful interpretation from partial image data by mimicking the human visual system. However, initialization of the surface is critical for the final result and its main drawbacks are very slow convergence and the huge number of iterations required. In this paper, we first show that the governing equation for the subjective surface flow can be rearranged in an AOS implementation, providing a near real-time solution to the shape completion problem in 2D and 3D. Then, we devise a new initialization paradigm where we first "condition" the viewpoint surface using the Fast-Marching algorithm. We compare the original method with our new algorithm on several examples of real 3D medical images, thus revealing the improvement achieved. PMID- 15794136 TI - Crest lines for surface segmentation and flattening. AB - We present a method for extracting feature curves called crest lines from a triangulated surface. Then, we calculate the geodesic Voronoi diagram of crest lines to segment the surface into several regions. Afterward, barycentric surface flattening using theory from graph embeddings is implemented and, using the Geodesic Voronoi diagram, we develop a faster surface flattening algorithm. PMID- 15794137 TI - On a construction of a hierarchy of best linear spline approximations using a finite element approach. AB - We present a method for the hierarchical approximation of functions in one, two, or three variables based on the finite element method (Ritz approximation). Starting with a set of data sites with associated function, we first determine a smooth (scattered-data) interpolant. Next, we construct an initial triangulation by triangulating the region bounded by the minimal subset of data sites defining the convex hull of all sites. We insert only original data sites, thus reducing storage requirements. For each triangulation, we solve a minimization problem: computing the best linear spline approximation of the interpolant of all data, based on a functional involving function values and first derivatives. The error of a best linear spline approximation is computed in a Sobolev-like norm, leading to element-specific error values. We use these interval/triangle/tetrahedron specific values to identify the element to subdivide next. The subdivision of an element with largest error value requires the recomputation of all spline coefficients due to the global nature of the problem. We improve efficiency by 1) subdividing multiple elements simultaneously and 2) by using a sparse-matrix representation and system solver. PMID- 15794138 TI - Point-based probabilistic surfaces to show surface uncertainty. AB - Efficient and informative visualization of surfaces with uncertainties is an important topic with many applications in science and engineering. In these applications, the correct course of action may depend not only on the location of a boundary, but on the precision with which that location is known. Examples include environmental pollution borderline detection, oil basin edge characterization, or discrimination between cancerous and healthy tissue in medicine. This paper presents a method for producing visualizations of surfaces with uncertainties using points as display primitives. Our approach is to render the surface as a collection of points and to displace each point from its original location along the surface normal by an amount proportional to the uncertainty at that point. This approaoh can be used in combination with other techniques such as pseudocoloring to produce efficient and revealing visualizations. The basic approach is sufficiently flexible to allow natural extensions; we show incorporation of expressive modulation of opacity, change of the stroke primitive, and addition of an underlying polygonal model. The method is used to visualize real and simulated tumor formations with uncertainty of tumor boundaries. The point-based technique is compared to pseudocoloring for a position estimation task in a preliminary user study. PMID- 15794139 TI - Haptics-based dynamic implicit solid modeling. AB - This paper systematically presents a novel, interactive solid modeling framework, Haptics-based Dynamic Implicit Solid Modeling, which is founded upon volumetric implicit functions and powerful physics-based modeling. In particular, we augment our modeling framework with a haptic mechanism in order to take advantage of additional realism associated with a 3D haptic interface. Our dynamic implicit solids are semi-algebraic sets of volumetric implicit functions and are governed by the principles of dynamics, hence responding to sculpting forces in a natural and predictable manner. In order to directly manipulate existing volumetric data sets as well as point clouds, we develop a hierarchical fitting algorithm to reconstruct and represent discrete data sets using our continuous implicit functions, which permit users to further design and edit those existing 3D models in real-time using a large variety of haptic and geometric toolkits, and visualize their interactive deformation at arbitrary resolution. The additional geometric and physical constraints afford more sophisticated control of the dynamic implicit solids. The versatility of our dynamic implicit modeling enables the user to easily modify both the geometry and the topology of modeled objects, while the inherent physical properties can offer an intuitive haptic interface for direct manipulation with force feedback. PMID- 15794140 TI - Simplification of three-dimensional density maps. AB - We consider scientific data sets that describe density functions over three dimensional geometric domains. Such data sets are often large and coarsened representations are needed for visualization and analysis. Assuming a tetrahedral mesh representation, we construct such representations with a simplification algorithm that combines three goals: the approximation of the function, the preservation of the mesh topology, and the improvement of the mesh quality. The third goal is achieved with a novel extension of the well-known quadric error metric. We perform a number of computational experiments to understand the effect of mesh quality improvement on the density map approximation. In addition, we study the effect of geometric simplification on the topological features of the function by monitoring its critical points. PMID- 15794141 TI - Confetti: object-space point blending and splatting. AB - In this paper, we present Confetti, a novel point-based rendering approach based on object-space point interpolation of densely sampled surfaces. We introduce the concept of a transformation-invariant covariance matrix of a set of points which can efficiently be used to determine splat sizes in a multiresolution point hierarchy. We also analyze continuous point interpolation in object-space and we define a new class of parameterized blending kernels as well as a normalization procedure to achieve smooth blending. Furthermore, we present a hardware accelerated rendering algorithm based on texture mapping and alpha-blending as well as programmable vertex and pixel-shaders. PMID- 15794142 TI - Digital art revolution. PMID- 15794143 TI - Applications of computers to dance. PMID- 15794144 TI - Tangible augmented interfaces for structural molecular biology. PMID- 15794145 TI - Artistic collaboration in designing VR visualizations. PMID- 15794146 TI - Automated eye motion using texture synthesis. PMID- 15794147 TI - A novel Monte Carlo noise reduction operator. PMID- 15794148 TI - Bounded blending for function-based shape modeling. PMID- 15794149 TI - A meshing scheme for efficient hardware implementation of butterfly subdivision using displacement mapping. PMID- 15794150 TI - Web-based shape modeling with HyperFun. PMID- 15794151 TI - Jabiru: harnessing Java 3D behaviors for device and display portability. PMID- 15794152 TI - Ten CAD challenges. PMID- 15794153 TI - Deriving linear transformations in 3D using Quaternion Algebra. PMID- 15794154 TI - Speed limit 55. PMID- 15794155 TI - Feature space interpretation of SVMs with indefinite kernels. AB - Kernel methods are becoming increasingly popular for various kinds of machine learning tasks, the most famous being the support vector machine (SVM) for classification. The SVM is well understood when using conditionally positive definite (cpd) kernel functions. However, in practice, non-cpd kernels arise and demand application in SVMs. The procedure of "plugging" these indefinite kernels in SVMs often yields good empirical classification results. However, they are hard to interpret due to missing geometrical and theoretical understanding. In this paper, we provide a step toward the comprehension of SVM classifiers in these situations. We give a geometric interpretation of SVMs with indefinite kernel functions. We show that such SVMs are optimal hyperplane classifiers not by margin maximization, but by minimization of distances between convex hulls in pseudo-Euclidean spaces. By this, we obtain a sound framework and motivation for indefinite SVMs. This interpretation is the basis for further theoretical analysis, e.g., investigating uniqueness, and for the derivation of practical guidelines like characterizing the suitability of indefinite SVMs. PMID- 15794156 TI - Light source position and reflectance estimation from a single view without the distant illumination assumption. AB - Several techniques have been developed for recovering reflectance properties of real surfaces under unknown illumination. However, in most cases, those techniques assume that the light sources are located at inifinity, which cannot be applied safely to, for example, reflectance modeling of indoor environments. In this paper, we propose two types of methods to estimate the surface reflectance property of an object, as well as the position of a light source from a single view without the distant illumination assumption, thus relaxing the conditions in the previous methods. Given a real image and a 3D geometric model of an object with specular reflection as inputs, the first method estimates the light source position by fitting to the Lambertian diffuse component, while separating the specular and diffuse components by using an iterative relaxation scheme. Our second method extends that first method by using as input a specular component image, which is acquired by analyzing multiple polarization images taken from a single view, thus removing its constraints on the diffuse reflectance property. This method simultaneously recovers the reflectance properties and the light source positions by optimizing the linearity of a log transformed Torrance-Sparrow model. By estimating the object's reflectance property and the light source position, we can freely generate synthetic images of the target object under arbitrary lighting conditions with not only source direction modification but also source-surface distance modification. Experimental results show the accuracy of our estimation framework. PMID- 15794157 TI - Vector-valued image regularization with PDEs: a common framework for different applications. AB - In this paper, we focus on techniques for vector-valued image regularization, based on variational methods and PDEs. Starting from the study of PDE-based formalisms previously proposed in the literature for the regularization of scalar and vector-valued data, we propose a unifying expression that gathers the majority of these previous frameworks into a single generic anisotropic diffusion equation. On one hand, the resulting expression provides a simple interpretation of the regularization process in terms of local filtering with spatially adaptive Gaussian kernels. On the other hand, it naturally disassembles any regularization scheme into the smoothing process itself and the underlying geometry that drives the smoothing. Thus, we can easily specialize our generic expression into different regularization PDEs that fulfill desired smoothing behaviors, depending on the considered application: image restoration, inpainting, magnification, flow visualization, etc. Specific numerical schemes are also proposed, allowing us to implement our regularization framework with accuracy by taking the local filtering properties of the proposed equations into account. Finally, we illustrate the wide range of applications handled by our selected anisotropic diffusion equations with application results on color images. PMID- 15794158 TI - Enhancing resolution along multiple imaging dimensions using assorted pixels. AB - Multisampled imaging is a general framework for using pixels on an image detector to simultaneously sample multiple dimensions of imaging (space, time, spectrum, brightness, polarization, etc.). The mosaic of red, green, and blue spectral filters found in most solid-state color cameras is one example of multisampled imaging. We briefly describe how multisampling can be used to explore other dimensions of imaging. Once such an image is captured, smooth reconstructions along the individual dimensions can be obtained using standard interpolation algorithms. Typically, this results in a substantial reduction of resolution (and, hence, image quality). One can extract significantly greater resolution in each dimension by noting that the light fields associated with real scenes have enormous redundancies within them, causing different dimensions to be highly correlated. Hence, multisampled images can be better interpolated using local structural models that are learned offline from a diverse set of training images. The specific type of structural models we use are based on polynomial functions of measured image intensities. They are very effective as well as computationally efficient. We demonstrate the benefits of structural interpolation using three specific applications. These are 1) traditional color imaging with a mosaic of color filters, 2) high dynamic range monochrome imaging using a mosaic of exposure filters, and 3) high dynamic range color imaging using a mosaic of overlapping color and exposure filters. PMID- 15794159 TI - Space-time super-resolution. AB - We propose a method for constructing a video sequence of high space-time resolution by combining information from multiple low-resolution video sequences of the same dynamic scene. Super-resolution is performed simultaneously in time and in space. By "temporal super-resolution," we mean recovering rapid dynamic events that occur faster than regular frame-rate. Such dynamic events are not visible (or else are observed incorrectly) in any of the input sequences, even if these are played in "slow-motion." The spatial and temporal dimensions are very different in nature, yet are interrelated. This leads to interesting visual trade offs in time and space and to new video applications. These include: 1) treatment of spatial artifacts (e.g., motion-blur) by increasing the temporal resolution and 2) combination of input sequences of different space-time resolutions (e.g., NTSC, PAL, and even high quality still images) to generate a high quality video sequence. We further analyze and compare characteristics of temporal super resolution to those of spatial super-resolution. These include: How many video cameras are needed to obtain increased resolution? What is the upper bound on resolution improvement via super-resolution? What is the temporal analogue to the spatial "ringing" effect? PMID- 15794160 TI - Salient closed boundary extraction with ratio contour. AB - We present ratio contour, a novel graph-based method for extracting salient closed boundaries from noisy images. This method operates on a set of boundary fragments that are produced by edge detection. Boundary extraction identifies a subset of these fragments and connects them sequentially to form a closed boundary with the largest saliency. We encode the Gestalt laws of proximity and continuity in a novel boundary-saliency measure based on the relative gap length and average curvature when connecting fragments to form a closed boundary. This new measure attempts to remove a possible bias toward short boundaries. We present a polynomial-time algorithm for finding the most-salient closed boundary. We also present supplementary preprocessing steps that facilitate the application of ratio contour to real images. We compare ratio contour to two closely related methods for extracting closed boundaries: Elder and Zucker's method based on the shortest-path algorithm and Williams and Thornber's method based on spectral analysis and a strongly-connected-components algorithm. This comparison involves both theoretic analysis and experimental evaluation on both synthesized data and real images. PMID- 15794161 TI - Recursive estimation of 3D motion and surface structure from local affine flow parameters. AB - A recursive structure from motion algorithm based on optical flow measurements taken from an image sequence is described. It provides estimates of surface normals in addition to 3D motion and depth. The measurements are affine motion parameters which approximate the local flow fields associated with near-planar surface patches in the scene. These are integrated over time to give estimates of the 3D parameters using an extended Kalman filter. This also estimates the camera focal length and, so, the 3D estimates are metric. The use of parametric measurements means that the algorithm is computationally less demanding than previous optical flow approaches and the recursive filter builds in a degree of noise robustness. Results of experiments on synthetic and real image sequences demonstrate that the algorithm performs well. PMID- 15794162 TI - A theoretical and experimental investigation of graph theoretical measures for land development in satellite imagery. AB - Today's commercial satellite images enable experts to classify region types in great detail. In previous work, we considered discriminating rural and urban regions [23]. However, a more detailed classification is required for many purposes. These fine classifications assist government agencies in many ways including urban planning, transportation management, and rescue operations. In a step toward the automation of the fine classification process, this paper explores graph theoretical measures over grayscale images. The graphs are constructed by assigning photometric straight line segments to vertices, while graph edges encode their spatial relationships. We then introduce a set of measures based on various properties of the graph. These measures are nearly monotonic (positively correlated) with increasing structure (organization) in the image. Thus, increased cultural activity and land development are indicated by increases in these measures-without explicit extraction of road networks, buildings, residences, etc. These latter, time consuming (and still only partially automated) tasks can be restricted only to "promising" image regions, according to our measures. In some applications our measures may suffice. We present a theoretical basis for the measures followed by extensive experimental results in which the measures are first compared to manual evaluations of land development. We then present and test a method to focus on, and (pre)extract, suburban-style residential areas. These are of particular importance in many applications, and are especially difficult to extract. In this work, we consider commercial IKONOS data. These images are orthorectified to provide a fixed resolution of 1 meter per pixel on the ground. They are, therefore, metric in the sense that ground distance is fixed in scale to pixel distance. Our data set is large and diverse, including sea and coastline, rural, forest, residential, industrial, and urban areas. PMID- 15794164 TI - Fast SVM training algorithm with decomposition on very large data sets. AB - Training a support vector machine on a data set of huge size with thousands of classes is a challenging problem. This paper proposes an efficient algorithm to solve this problem. The key idea is to introduce a parallel optimization step to quickly remove most of the nonsupport vectors, where block diagonal matrices are used to approximate the original kernel matrix so that the original problem can be split into hundreds of subproblems which can be solved more efficiently. In addition, some effective strategies such as kernel caching and efficient computation of kernel matrix are integrated to speed up the training process. Our analysis of the proposed algorithm shows that its time complexity grows linearly with the number of classes and size of the data set. In the experiments, many appealing properties of the proposed algorithm have been investigated and the results show that the proposed algorithm has a much better scaling capability than Libsvm, SVMlight, and SVMTorch. Moreover, the good generalization performances on several large databases have also been achieved. PMID- 15794163 TI - Statistical shape analysis: clustering, learning, and testing. AB - Using a differential-geometric treatment of planar shapes, we present tools for: 1) hierarchical clustering of imaged objects according to the shapes of their boundaries, 2) learning of probability models for clusters of shapes, and 3) testing of newly observed shapes under competing probability models. Clustering at any level of hierarchy is performed using a mimimum variance type criterion criterion and a Markov process. Statistical means of clusters provide shapes to be clustered at the next higher level, thus building a hierarchy of shapes. Using finite-dimensional approximations of spaces tangent to the shape space at sample means, we (implicitly) impose probability models on the shape space, and results are illustrated via random sampling and classification (hypothesis testing). Together, hierarchical clustering and hypothesis testing provide an efficient framework for shape retrieval. Examples are presented using shapes and images from ETH, Surrey, and AMCOM databases. PMID- 15794165 TI - An evaluation of multimodal 2D+3D face biometrics. AB - We report on the largest experimental study to date in multimodal 2D+3D face recognition, involving 198 persons in the gallery and either 198 or 670 time lapse probe images. PCA-based methods are used separately for each modality and match scores in the separate face spaces are combined for multimodal recognition. Major conclusions are: 1) 2D and 3D have similar recognition performance when considered individually, 2) combining 2D and 3D results using a simple weighting scheme outperforms either 2D or 3D alone, 3) combining results from two or more 2D images using a similar weighting scheme also outperforms a single 2D image, and 4) combined 2D+3D outperforms the multiimage 2D result. This is the first (so far, only) work to present such an experimental control to substantiate multimodal performance improvement. PMID- 15794166 TI - Edge and corner detection by photometric quasi-invariants. AB - Feature detection is used in many computer vision applications such as image segmentation, object recognition, and image retrieval. For these applications, robustness with respect to shadows, shading, and specularities is desired. Features based on derivatives of photometric invariants, which we will call full invariants, provide the desired robustness. However, because computation of photometric invariants involves nonlinear transformations, these features are Instable and, therefore, impractical for many applications. We propose a new class of derivatives which we refer to as quasi-invariants. These quasi invariants are derivatives which share with full photometric invariants the property that they are insensitive for certain photometric edges, such as shadows or specular edges, but without the inherent instabilities of full photometric invariants. Experiments show that the quasi-invariant derivatives are less sensitive to noise and introduce less edge displacement than full invariant derivatives. Moreover, quasi-invariants significantly outperform the full invariant derivatives in terms of discriminative power. PMID- 15794167 TI - Generalized mosaicing: polarization panorama. AB - We present an approach to image the polarization state of object points in a wide field of view, while enhancing the radiometric dynamic range of maging systems by generalizing image mosaicing. The approach is biologically-inspired, as it emulates spatially varying polarization sensitivity of some animals. In our method, a spatially varying polarization and attenuation filter is rigidly attached to a camera. As the system moves, it senses each scene point multiple times, each time filtering it through a different filter polarizing angle, polarizance, and transmittance. Polarization is an additional dimension of the generalized mosaicing paradigm, which has recently yielded high dynamic range images and multispectral images in a wide field of view using other kinds of filters. The image acquisition is as easy as in traditional image mosaics. The computational algorithm can easily handle nonideal polarization filters (partial polarizers), variable exposures, and saturation in a single framework. The resulting mosaic represents the polarization state at each scene point. Using data acquired by this method, we demonstrate attenuation and enhancement of specular reflections and semireflection separation in an image mosaic. PMID- 15794169 TI - An EM algorithm for the block mixture model. AB - Although many clustering procedures aim to construct an optimal partition of objects or, sometimes, of variables, there are other methods, called block clustering methods, which consider simultaneously the two sets and organize the data into homogeneous blocks. Recently, we have proposed a new mixture model called block mixture model which takes into account this situation. This model allows one to embed simultaneous clustering of objects and variables in a mixture approach. We have studied this probabilistic model under the classification likelihood approach and developed a new algorithm for simultaneous partitioning based on the Classification EM algorithm. In this paper, we consider the block clustering problem under the maximum likelihood approach and the goal of our contribution is to estimate the parameters of this model. Unfortunately, the application of the EM algorithm for the block mixture model cannot be made directly; difficulties arise due to the dependence structure in the model and approximations are required. Using a variational approximation, we propose a generalized EM algorithm to estimate the parameters of the block mixture model and, to illustrate our approach, we study the case of binary data by using a Bernoulli block mixture. PMID- 15794168 TI - Geometric and algebraic constraints of projected concentric circles and their applications to camera calibration. AB - We investigate the projective properties of the feature consisting of two concentric circles. We demonstrate there exist geometric and algebraic constraints on its projection. We show how these constraints greatly simplify the recoveries of the affine and Euclidean structures of a 3D plane. As an application, we assess the performances of two camera calibration algorithms. PMID- 15794170 TI - A hybrid neural network system for pattern classification tasks with missing features. AB - A hybrid neural network comprising Fuzzy ARTMAP and Fuzzy C-Means Clustering is proposed for pattern classification with incomplete training and test data. Two benchmark problems and a real medical pattern classification task are employed to evaluate the effectiveness of the hybrid network. The results are analyzed and compared with those from other methods. PMID- 15794171 TI - Levetiracetam in pediatrics. AB - Levetiracetam is a new antiepileptic drug whose efficacy and tolerability are already well known in adults. Few studies are available in children. This review, based on the international literature, aims to identify and make known the possible indications for levetiracetam in childhood. Most studies suggest that levetiracetam is effective against partial and generalized epilepsy. In resistant partial epilepsy, the percentage of responders reaches 64%, with 8 to 23% seizure free. Levetiracetam is used to treat symptomatic and idiopathic epilepsies. The drug has also proven effective against photosensitivity and epileptic and nonepileptic myoclonus. The most frequent side effects involve the behavioral sphere and manifest mostly in patients with a history of behavioral problems. In some patients, levetiracetam increases the number of seizures, but this adverse reaction can be partially avoided with slow titration. Doses for children should be 130 to 140% of those advised for adults. Levetiracetam seems to have a broad spectrum of action and is, on the whole, well tolerated. Its efficacy against generalized epilepsy is particularly promising in childhood. PMID- 15794172 TI - Congenital myopathies in the new millennium. AB - Few medical disciplines have benefited so enormously from the molecular revolution as myology. Whereas the congenital myopathies have flourished from enzyme histochemistry and electron microscopy, defining individual congenital myopathies by structural abnormalities, genetic research has only recently focused on congenital myopathies. However, a number of congenital myopathies have been molecularly elucidated: central and multiminicore diseases, nemaline myopathy, myotubular myopathy, and congenital myopathy marked by aggregation of proteins, giving rise to the concept of protein aggregate myopathies, to which now desminopathies, alpha-B crystallinopathies, selenoproteinopathy, myotilinopathy, actinopathies, and myosinopathies belong. Based on recent identification of mutations in respective genes, the principle "from morphology, that is, immunohistochemistry, to molecular analysis" through recognition of certain accrued proteins within muscle fibers and subsequent analysis of their respective genes has resulted in a wealth of genetic data and in reconsidering classification and nosologic interpretation of certain congenital myopathies. This heuristic principle needs to be further applied to other genetically still obscure congenital myopathies. PMID- 15794173 TI - Cognitive deficits in children with sickle cell disease. AB - We tested a hypothesis that children with sickle cell disease who are completely normal by magnetic resonance imaging can still be cognitively impaired, as predicted by a model of diffuse brain injury. Fifty-four patients with hemoglobin SS (average age 10.9 years +/- 2.9 years SD) were examined with the Wechsler Intelligence Scale for Children-III (WISC-III) and were randomly matched by age, race, and gender with healthy children from the Wechsler normative database. Patients were also imaged at 1.5 Tesla with standard imaging sequences. Among 30 patients who were normal by magnetic resonance imaging, there were substantial deficits in Wechsler Full-Scale IQ, Verbal IQ, and Performance IQ (all P < .01) compared with African-American controls. The patient Wechsler Full-Scale IQ was 12.9 points lower than that of controls and decreased as a function of age (probability = .014). The findings suggest that there is diffuse brain injury in patients and that patient deficits increase with age. PMID- 15794174 TI - ThOMAs: the other means of assessment. AB - We describe a pilot study to investigate whether drawing "Thomas the Tank Engine" could be as effective a measure of developmental progress as the Goodenough Harris Draw A Man test against the ThOMAs test (The Other Means of Assessment), with internal validation. The study included 95 children aged between 3 and 11 years of age, including a subgroup of 13 children with registered special needs from community and general pediatric clinics within Birmingham, UK, as a means of validation. There was no significant evidence that ThOMAS was either culturally or sex biased. Using regression analysis, nine items were found to correlate highly with actual age, and their total score gave a correlation of 0.563 with age. Adding further items did not increase this. After being converted into age standardized scores, ThOMAS was as sensitive and specific as the Draw A Man test, and more so above a defined age-standardized threshold. This pilot study suggests that drawing Thomas the Tank Engine would appear to be as sensitive and specific a means of identifying children with special needs as the Goodenough-Harris Draw A Man test. The relatively small sample size means that further research is necessary to further define the age standardizations and to refine the ThOMAs test. PMID- 15794175 TI - Add-on melatonin improves sleep behavior in children with epilepsy: randomized, double-blind, placebo-controlled trial. AB - This double-blind, randomized, placebo-controlled study in epileptic children, aged 3 to 12 years, evaluated the effect of add-on melatonin on the sleep behavior of these children on sodium valproate monotherapy using a parental questionnaire. Of the 31 patients, 16 randomly received add-on melatonin, whereas 15 received add-on placebo. The questionnaire showed good internal consistency in our patient population (Cronbach's alpha = .83). The percentage decrease in the median total sleep score was 24.4 (range 0.0-34.9) in the valproate + melatonin group compared with 14.0 (range -2.2-18.8) in the valproate + placebo group, the difference being statistically significant (P < .05). The median percentage decrease in the parasomnias score was 60 (range 0.0-70.8) in the valproate + melatonin group compared with 36.4 (range 0.0-63.2) in the valproate + placebo group, the difference being statistically significant (P < .05). There was no significant difference between the percentage decrease in the daytime drowsiness scores and sleep fragmentation scores. Parent-child interaction subscale scores were not significantly different between age groups. The age at onset of seizures and the type of seizures did not correlate significantly to the total sleep scores. Given that sleep problems are known to complicate epilepsy, add-on melatonin, which has a wide safety window, can be of promise in the pharmacotherapy of pediatric epilepsy. PMID- 15794176 TI - Episodic hyponatremia in mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes (MELAS). AB - Various organ involvements and endocrinologic abnormalities associated with electrolyte imbalance, including hyponatremia, are seen in patients with mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes (MELAS); however, the clinical significance of hyponatremia in these patients is rarely reported. We analyzed the serum sodium concentration profiles and clinical and laboratory data of seven patients with MELAS. We found that hyponatremia occurred episodically in four of the seven patients from an early stage of the disease. We identified excessive sodium loss in urine to be associated with the hyponatremic episodes and the causes of hyponatremia in two patients as relative adrenal insufficiency, acute renal failure, and serious paralytic ileus. However, even extensive examinations failed to reveal the cause in other patients. Because severe hyponatremia can cause serious complications, clinicians should pay attention to serum sodium levels and maintain them properly in patients with MELAS. PMID- 15794177 TI - Botulinum toxin injections for children with excessive drooling. AB - The objective of this study was to evaluate the feasibility of ultrasonography guided injections of botulinum toxin A into the parotid glands of children with severe drooling (sialorrhea). Excessive drooling is common in children with chronic neurologic disorders. Preliminary observations in adults suggest that injections of botulinum toxin A into the parotid glands can decrease drooling, but the optimal dose, sites of injection, and concomitant use of imaging during injections and its use for children have not been established. Ultrasonography was used to guide the injection of botulinum toxin (10-25 IU) into both parotid glands of nine children with excessive drooling. Subjective and objective measures of the severity of drooling were collected before and after botulinum toxin A injections. A booster injection was provided if the initial response was inadequate. Injections were well tolerated, and no adverse reactions were observed. Ultrasonography revealed that the parotid gland showed a variable depth, extent, and vascularization. Eight of nine patients needed a booster injection after 1 month. Objective measures of drooling severity were improved in seven of nine patients. However, subjective improvement was reported in only three of nine patients, and this improvement was functionally significant in only one patient. Although intraparotid injection of botulinum toxin A is safe and causes a reduction in saliva production in children, the doses used in this study did not result in functionally significant improvement. Higher doses of botulinum toxin A in the parotid glands or concomitant injections into the submandibular glands can increase the efficacy of these injections. Variability in size, depth, and vascular supply of the parotid gland suggests the importance of ultrasonography guidance for optimizing injections. These results underscore the need for further studies to establish the efficacy of this treatment in children. PMID- 15794178 TI - Study of multimodal evoked potentials in patients with type 1 Gaucher's disease. AB - To detect early subclinical nervous dysfunction in Gaucher's disease type 1, we carried out motor, brainstem auditory, visual, and somatosensory evoked potentials in 17 patients with Gaucher's disease type 1. Central motor evoked potential abnormalities were found in nine patients (69.2%), consisting of an increased motor threshold in all, with prolonged central motor conduction time in two patients. Brainstem auditory evoked potentials were abnormal in five patients (31.2%), and the most frequent abnormality was a bilateral increased I-III interpeak latency. Visual evoked potentials showed a delayed latency of the P100 wave in four patients (25%). Somatosensory evoked potential abnormalities were found in three patients (18.7%), consisting of an increased N13-N20 interval in two patients and a not reproducible N13 wave in one patient. Our findings suggest that the multimodal evoked potential approach provides information about nervous subclinical damage in Gaucher's disease type 1; transcranial magnetic stimulation proved to be the most sensitive tool. Early detection of subclinical neurologic dysfunction can be useful in view of more effective therapeutic strategies. PMID- 15794179 TI - Cognitive changes in children treated for acute lymphoblastic leukemia with chemotherapy only according to the Pediatric Oncology Group 9605 protocol. AB - The purpose of this study was to examine cognitive functioning and neuroimaging in children with leukemia treated with the Pediatric Oncology Group 9605 protocol at the Children's Hospital of Eastern Ontario. Mean age at diagnosis was 4.88 +/- 2.54 years. The mean (n = 24) Wechsler Verbal and Performance IQ fell in the low average range (87.33 +/- 15.69 and 84.83 +/- 19.11, respectively). Mean (n = 20) Verbal and Visual Memory Indexes of 82.95 +/- 15.46 and 88.30+/- 10.80, respectively, were obtained. The proportion of scores on measures of intelligence and memory falling > 1 SD below the normative mean was substantially higher than expected. Paired t-test suggested that Wechsler Verbal IQ and memory remained stable, whereas Wechsler Performance IQ declined significantly. The results of growth curve analyses replicated these findings and suggested a significant adverse effect of cumulative dosage of intrathecal methotrexate on estimated Wechsler Performance IQ. Although only two children experienced seizures, 78% of the group showed leukoencephalopathy on at least one magnetic resonance image. Reliance on seizures as a predictor of leukoencephalopathy might underestimate the incidence of neurotoxicity. PMID- 15794180 TI - Behavioral characteristics of Prader-Willi syndrome in Korea: comparison with children with mental retardation and normal controls. AB - The purpose of this study was to examine the psychiatric characteristics of children with Prader-Willi syndrome in Korea, focusing particularly on their behavioral problems and obsessive-compulsive spectrum symptoms. Fourteen patients with Prader-Willi syndrome, together with their parents, underwent a psychiatric interview and parent questionnaire consisting of a Child Behavior Checklist. Twenty-four patients with mental retardation and 45 normal students were selected as control groups. Compared with the normal control group, the rates of inclusion in the clinical range and the mean scores with regard to social problems, thought problems, attention problems, delinquent behavior, aggressive behavior, externalizing problems, and total problems profiles were significantly higher (P < .01) in the Prader-Willi syndrome group according to the results of the Child Behavior Checklist. Compared with the mental retardation group, there was a statistically significant (P < .05) difference in the delinquent behavior profile. Comparing selected Child Behavior Checklist items related to obsessive compulsive spectrum symptoms, the Prader-Willi syndrome group showed significantly more (P < .05) compulsion, skin picking, and stealing than the mental retardation or normal control groups. These findings suggest that children with Prader-Willi syndrome in Korea have many behavioral problems, including obsessive-compulsive spectrum symptoms, needing proper psychiatric attention and treatment. This is the first study in Korea to evaluate the psychiatric and behavioral characteristics of children with Prader-Willi syndrome. PMID- 15794181 TI - Cerebellar injury in the extremely premature infant: newly recognized but relatively common outcome. AB - Severe injury to the cerebellum as a complication of extreme prematurity with extremely low birthweight was recently described in 13 children with the clinical diagnosis of cerebral palsy. We report another 10 cases of this syndrome. The clinical features include striking motor impairment and variable degrees of ataxia and athetosis or dystonia, which represent a distinct clinical type of cerebral palsy. Most are severely damaged, with cognitive, language, and motor delays. All are microcephalic, except one with hydrocephalus. Neuroimaging studies demonstrate the absence of major portions of the cerebellum involving both the inferior vermis and hemispheres. Most also have injury of a less severe nature in the cerebrum. This report indicates that this is not an uncommon outcome of extremely low birthweight infants, and we hope to encourage further investigations into the relative frequency and likely etiologies of the condition. PMID- 15794182 TI - Clinical and genetic features in two families with MELAS and the T3271C mutation in mitochondrial DNA. AB - The majority of patients with MELAS (mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes) have the A3243G point mutation. The much rarer T3271C mutation has been reported predominantly in Japanese subjects. Our objective was to better define the clinical phenotype and mutation load in patients with MELAS and the T3271C mutation in mitochondrial DNA. We present clinical and molecular genetic data in two pedigrees with the T3271C mutation. The age at onset was 8 years in one proband and 14 years in the other. Both patients had migrainelike headache, seizures, and strokelike episodes. Mutation loads were quantified in multiple tissues from the patients and from family members by polymerase chain reaction-restriction fragment length polymorphism analysis. The symptoms in both probands were typical of MELAS, and, contrary to previous reports, onset was early. Hearing loss was less common than in typical MELAS, and ragged red fibers were absent. The proportion of mutant genomes was consistently and markedly greater in DNA from urinary sediment than from blood. In the mother of one proband, mutant genomes were detected only in DNA from hair follicles and cheek mucosa The phenotype of patients with the T3271C mutation might not be as distinct as that of the A3243G mutation, as previously described. Our data also suggest that urine is a better source of DNA than blood for diagnosis and that multiple tissues should be studied in maternal relatives, especially when the mutation cannot be detected in blood. PMID- 15794183 TI - Type I spinal muscular atrophy can mimic sensory-motor axonal neuropathy. AB - Spinal muscular atrophy is a group of allelic autosomal recessive disorders characterized by progressive motoneuron loss, symmetric weakness, and skeletal muscle atrophy. It is traditionally considered a pure lower motoneuron disorder, for which a current definitive diagnosis is now possible by molecular genetic testing. We report two newborns with a clinical phenotype consistent with that of spinal muscular atrophy type I and nerve conduction studies and electromyography suggesting more extensive sensory involvement than classically described with spinal muscular atrophy. Molecular testing confirmed spinal muscular atrophy in patient 1 but not in patient 2. Thus, in the setting of a suspected congenital axonal neuropathy, molecular testing might be necessary to distinguish spinal muscular atrophy type I from infantile polyneuropathy. PMID- 15794184 TI - Valproate-induced eosinophilia in children with epilepsy: role of interleukin-5. AB - Interleukin-5 contributes both in eosinophilopoiesis and neural development. Serum interleukin-5 levels were measured with enzyme-linked immunosorbent assay technique in 68 children with epilepsy receiving sodium valproate monotherapy and compared with the levels of 60 healthy controls and 14 children with epilepsy receiving carbamazepine. Eosinophilia was observed in 35.3% of children receiving valproate. Interleukin-5 in valproate users was significantly higher compared with children receiving carbamazepine and controls. Valproate users who exhibited eosinophilia had higher interleukin-5 levels compared with those without eosinophilia. However, the interleukin-5 level was also elevated, although to a lesser degree, in children without eosinophilia. The majority of valproate responders had high interleukin-5 levels. A positive correlation between interleukin-5 levels and the eosinophil count was also noted. We postulate that valproate contributes to the pathogenesis of eosinophilia, probably inducing interleukin-5 production. The finding that serum interleukin-5 was significantly elevated in valproate responders and even in valproate users without eosinophilia suggests that the increase in interleukin-5 might represent one of valproate's antiepileptic mechanisms. PMID- 15794185 TI - Postural headache in a child with Marfan syndrome: case report and review of the literature. AB - The case of a 10-year-old female with Marfan syndrome and postural headache secondary to spontaneous intracranial hypotension is described. The patient was found to have multiple dural ectasias and a cerebrospinal fluid leak at the left cervicothoracic junction. Her presentation, diagnostic work-up, and management are reviewed, and the relevant literature is discussed. Spontaneous intracranial hypotension secondary to cerebrospinal fluid leaks from dural ectasia should be recognized as a potential complication in children with Marfan syndrome and other connective tissue diseases. PMID- 15794186 TI - Molybdenum cofactor deficiency presenting with severe metabolic acidosis and intracranial hemorrhage. AB - Molybdenum cofactor deficiency leads to combined deficiency of sulfite oxidase, xanthine dehydrogenase, and aldehyde oxidase enzyme activities. The major clinic symptom is intractable seizures seen soon after birth. No definite therapy is available. We report here a newborn with molybdenum cofactor deficiency associated Dandy-Walker malformation who presented with severe lactic acidosis and intracranial hemorrhage. PMID- 15794187 TI - Topiramate-valproate-induced hyperammonemic encephalopathy syndrome: case report. AB - A 15-year-old boy with inverted duplication of chromosome 15 was admitted for acute onset of irritability, increasing sleepiness, and worsening of seizures. He had been on valproate and other anti-convulsants. However, he was found to have hyperammonemia within 2 weeks after the addition of low-dose topiramate to valproate. He recovered within 7 days after discontinuation of valproate. Topiramate was tailed off. The reintroduction of valproate monotherapy caused hyperammonemia again without clinical features of encephalopathy. He also developed anticonvulsant hypersensitivity syndrome following the use of phenytoin. We propose the term topiramate-valproate-induced hyperammonemic encephalopathy syndrome to include the following features: excessive sleepiness or somnolence, aggravation of seizures, hyperammonemia, and absence of triphasic waves on electroencephalography in any individual on simultaneous topiramate valproate therapy. The ammonia level ranged from 1.5 to 2 times normal. The serum valproate level might be within the therapeutic range. The possible mechanism is topiramate-induced aggravation of all the known complications of valproate monotherapy. This condition is reversible with cessation of either valproate or topiramate. PMID- 15794188 TI - Severe epilepsy and pachygyria associated with peculiar facial traits characterize Fryns-Aftimos syndrome. AB - We describe an 18-year-old boy with a frontofacial syndrome. The presence of severe epilepsy led to a neuroimaging investigation, which revealed a diffuse pachygyric (normal thickness and simplified folding) cortex that was bilateral and affected all supratentorial regions, sparing the medial and inferior aspects of the frontal lobe plus the posterior occipital region. In addition, an unusual skull base presentation with small posterior fossa, narrowed foramen magnum, and low torcular and tentorial insertions was documented. Dysmorphic ventricles and enlarged cavi of the septi pellucidum, vergae, and velum interpositum were also noted. This case shares many similarities with two patients formerly described by Fryns and Aftimos, who were previously classified as having mental retardation and multiple congenital anomalies syndrome. The existence of a new case corroborates the idea of a distinct condition with particular characteristic dysmorphic facial features and the presence of a specific malformation of cortical development associated with severe epilepsy. We believe that epilepsy and pachygyria play a central role in distinguishing this syndrome from others classified as frontofacial or cerebrofrontofacial syndromes, and we propose the term "Fryns-Aftimos syndrome" to name it. PMID- 15794189 TI - Childhood acute disseminated encephalomyelitis presenting as Foix-Chavany-Marie syndrome. AB - A 10-year-old boy presented with anarthria and bilateral central faciolinguovelopharyngeomasticatory paralysis with "automatic voluntary dissociation" suggestive of Foix-Chavany-Marie syndrome or anterior opercular syndrome following a nonspecific upper respiratory infection. Brain magnetic resonance imaging revealed bilateral subcortical perisylvian demyelination in addition to multiple subcortical white-matter demyelination. The clinicoradiologic picture suggested acute disseminated encephalomyelitis. The child recovered completely following intravenous methylprednisolone and intravenous immunoglobulin therapy. This is the first report of anterior opercular syndrome in childhood acute disseminated encephalomyelitis. PMID- 15794190 TI - Paroxysmal tonic upgaze and partial tetrasomy of chromosome 15: a novel genetic association. AB - Paroxysmal tonic upgaze of childhood is an eye movement abnormality characterized by periodic episodes of conjugate upward eye deviation. Although the spectrum of paroxysmal tonic upgaze has broadened considerably, a specific pathophysiology has not been elucidated. We report an infant with paroxysmal tonic upgaze who presented to his pediatrician with associated hypotonia and gross motor delay. High-resolution chromosome analysis demonstrated a supernumerary dicentric bisatellited marker chromosome. Analysis revealed partial tetrasomy of 15q. Given the significant amount of euchromatin demonstrated, it is likely that the chromosomal abnormalities are causative for the constellation of manifestations in this patient. To date, a specific genetic abnormality has not been associated with paroxysmal tonic upgaze. The finding of a genetic association with paroxysmal tonic upgaze might help characterize the substrate for ophthalmologic manifestations. Further study in this chromosomal region in patients with paroxysmal tonic upgaze is warranted. PMID- 15794192 TI - Extreme microcephaly with agyria-pachygyria, partial agenesis of the corpus callosum, and pontocerebellar dysplasia. AB - A 1-year-old boy with extreme microcephaly and a complex brain malformation is reported. Magnetic resonance imaging revealed an abnormal gyral pattern with features of the agyria-pachygyria spectrum, partial agenesis of the corpus callosum, severely hypoplastic posterior cerebellar vermis, and an abnormal foliation pattern of the cerebellar hemispheres associated with a flat and wide isthmus and pons. Although this phenotype shares some features with malformations classified as microcephaly with a simplified gyral pattern, microlissencephaly, or lissencephaly with cerebellar hypoplasia, none of the several subgroups of these categories are identical to the cerebral dysgenesis found in this patient. PMID- 15794191 TI - Recurrent meningitis associated with a petrous apex cephalocele. AB - We present the case of a 6-year-old girl with recurrent bacterial meningitis and cerebrospinal fluid (CSF) rhinorrhea associated with a petrous apex cephalocele (PAC). We diagnosed her by means of three-dimensional computed tomography (CT) and heavily T2-weighted magnetic resonance imaging (MRI). Petrous apex cephaloceles are usually an asymptomatic incidental finding in adults; however, they should be considered as a possible cause of CSF rhinorrhea, otorrhea, and recurrent meningitis in children. PMID- 15794193 TI - The systemic nature of the antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS, Hughes' syndrome) is a systemic autoimmune disorder characterized by arterial and/or venous thrombosis and recurrent foetal loss, accompanied by mild to moderate thrombocytopaenia and elevated titres of antiphospholipid antibodies (aPLs): lupus anticoagulant (LAC) and/or anticardiolipin (aCL) antibodies. APS was defined originally in 1983 in systemic lupus erythematosus (SLE) patients, but later it was found that APS can be primary or secondary to other autoimmune diseases or malignancy. During the past 20 years many organs have been reported to be involved in this syndrome and the clinical manifestations are seen in every medical field. Moreover, many aPLs have been found in APS besides aCLs and LACs, which bind to the autoantigen beta-2 glycoprotein I (beta2GPI). Treatment for APS, based on antiplatelet and anticoagulation drugs, is dependent on various parameters, including whether SLE is also present, classical vs non-classical manifestations of the diseases, women with APS based on pregnancy morbidity, the presence of elevated aCL antibody titres in the absence of clinical manifestations, and catastrophic APS. PMID- 15794194 TI - Which factors are related to the presence of atherosclerosis in rheumatoid arthritis? AB - OBJECTIVE: An accelerated progression of atherosclerosis may contribute to the increased mortality due to cardiovascular disease reported in rheumatoid arthritis (RA). The aim of this study was to identify variables, related to disease onset as well as to disease progression, of importance for the presence of atherosclerosis, as diagnosed by B-mode ultrasonography, in patients with medium-term RA. The results are based on the co-analysis of retrospective data as well as cross-sectional data. The impact of RA per se on atherosclerosis was evaluated relative to age- and sex-matched controls. METHODS: Thirty-nine RA patients, with a maximum age of 65 years, who had previously been included in a large retrospective cohort study, were assessed by duplex scanning after a disease duration of 19-23 years. In the present study, factors identified in the two earlier studies were assessed for their potential relationship with intima media wall thickness (IMT) of the common carotid artery (CCA), and the presence and grade of atherosclerotic plaques of the CCA and the common femoral artery, in regression models. The candidate co-variates were: variables reflecting inflammatory activity at disease onset and at the time of ultrasound assessment, established cardiovascular risk factors, pharmacological treatment [corticosteroids, disease-modifying anti-rheumatic drugs (DMARDs)], and the presence of complications and co-morbidity identified during disease progression, as well as lipid levels, anti-lipid antibodies, haemostatic factors, and markers of immune activation measured at ultrasound assessment. RESULTS: In patients with RA, analysis of simple linear regression models revealed those variables significantly associated with IMT-CCA to be age, tissue plasminogen activator (tPA) antigen, cholesterol, low density lipoprotein (LDL)-cholesterol, triglycerides, and atherosclerotic plaques while neither inflammatory status at disease onset, traditional cardiovascular risk factors, or pharmacological treatment during disease had any significant impact on IMT. In an estimated multiple linear regression model, variables associated with increasing log of IMT CCA were the log of cholesterol and of soluble intracellular adhesion molecule 1 (sICAM-1), while methotrexate treatment tended to have a decreasing effect. In simple binary logistic regression, atherosclerotic plaques were associated with age, IMT-CCA, smoking, and the levels of sICAM-1, sE-selectin, interleukin-2 soluble receptor alpha (IL-2sRalpha), plasminogen activator inhibitor-1 (PAI-1) mass, cholesterol, LDL-cholesterol, and the LDL/high density lipoprotein (HDL) ratio. A multiple approach indicated that plaques were associated with age, cholesterol, and sE-selectin. Severe plaques were associated with LDL-cholesterol and disease duration. Logistic regression in the age- and sex-matched case control study revealed that IMT-CCA was, together with the D-dimer, associated with RA per se. CONCLUSION: Levels of lipids and adhesion molecules were associated with the presence of atherosclerosis in RA. IMT-CCA was associated with RA per se. Disease duration could predict severe atherosclerotic plaques. Treatment with methotrexate seemed to decrease the IMT-CCA. PMID- 15794195 TI - Clinical and radiological disease-course in a Swedish DMARD-treated early RA inception cohort: an observational study. AB - OBJECTIVES: Disease-modifying anti-rheumatic drugs (DMARDs) decrease clinical signs and symptoms in rheumatoid arthritis (RA). However, radiographic changes sometimes continue to accrue despite effective suppression of clinical symptoms by therapy. The objective of this study was to identify whether successful clinical disease-control in a Swedish early RA-inception cohort of patients led to an attenuation of radiological progression. PATIENTS AND METHODS: We analysed clinical data and radiographs of 95 patients who were on a stable treatment regimen [methotrexate (MTX), sulfasalazine (SSZ), oral gold (AUR)] or who had changed between different DMARDs during the 2-year observation period [multiple therapy failures (mTF)]. Radiographs were quantified using the modified Larsen score and 'X-Ray RheumaCoach' software. RESULTS: Clinical measures improved markedly (p <0.001) from baseline to year 2 under AUR, MTX, and SSZ therapy but not in the mTF group. Similar levels of disease control were seen for each DMARD. During this period, patients treated with AUR had a deltaLarsen score (+ 14.5+/ 1.3) similar to mTF patients (+ 15.8+/-1.1) but greater than patients on MTX (+8.6+/-0.8) or SSZ (+9.1+/-0.8). CONCLUSIONS: This study confirms that radiological progression occurs despite a clinically acceptable disease control, but also shows that, given the same degree of clinical disease control, radiological progression can be different for different DMARDs. PMID- 15794196 TI - Plasma TNF binding capacity profiles during treatment with etanercept in rheumatoid arthritis. AB - BACKGROUND: Etanercept (Enbrel) induces a rapid and sustained decline in disease activity in the majority of patients with refractory rheumatoid arthritis (RA). In these patients neutralization of TNFalpha and lymphotoxin (LT), previously termed TNFbeta is mediated by etanercept itself, as well as by naturally occurring soluble TNF receptors. However, the clinical response to treatment with etanercept may vary. Previously, pharmacokinetic studies have focused on the molar concentrations of etanercept, but very little is known about the kinetics of bioactive etanercept in patients treated with etanercept. The purpose of this study was to evaluate kinetics, including inter- and intraindividual variations of the total TNF binding capacity, in RA patients who were on a standard treatment schedule with etanercept. METHODS: Plasma samples were collected daily from 16 RA patients who were in the steady-state phase during treatment with etanercept 25 mg subcutaneous once (n = 2) or twice (n = 14) weekly. The inflammatory activity, including Health Assessment Questionnaire (HAQ) score, numbers of painful and swollen joints, and ESR, was assessed at inclusion; CRP was measured on a daily basis. The samples were incubated with human recombinant LT to a concentration of 1000 pg/mL and the levels of detectable LT were measured by ELISA specific for free LT. The LT binding capacity (LTBC) was expressed in arbitrary units (AU) as the percentage value of bound LT to added LT. RESULTS: The median LTBC values measured during the treatment schedule from Day 1 (before the injection) to Day 4 (before the next injection) were 47 AU. The LTBC values in each individual patient generally remained fairly stable through the treatment schedule, and there were no significant differences in LTBC levels in samples obtained on a daily basis during the treatment schedule. However, a pronounced variation between the patients was noticed with LTBC values ranging from 10-82 AU (coefficient of variation=38%). No significant association was found between LTBC levels and clinical measures of disease activity, including HAQ and numbers of swollen or painful joints. However, among patients with high LTBC levels (> or =65 AU), elevated levels of ESR and CRP were less frequent (0%) compared with patients who had lower LTBC levels, in which the frequencies of elevated ESR and CRP were 53% and 37%, respectively. Soluble TNF receptor 1 (sTNFR1) remained stable between the injections and correlated with the number of swollen joints, but did not correlate with LTBC values. CONCLUSION: LTBC levels appeared stable in each individual RA patient who was on the standard treatment schedule with weekly injections of etanercept, but the inter-individual variations were considerable. PMID- 15794197 TI - TGF-beta1 polymorphism determines the progression of joint damage in rheumatoid arthritis. AB - OBJECTIVE: To determine whether transforming growth factor-beta1 (TGF-beta1) polymorphisms are associated with susceptibility to rheumatoid arthritis (RA) and to analyse whether they can affect the progression of radiographic severity. METHODS: A total of 143 RA patients and 148 healthy unrelated controls were tested for the TGF-beta1 polymorphisms using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). RESULTS: The TGF-beta1 polymorphisms were not associated with susceptibility to RA, although there was a trend that -509C/T and the 869T/C polymorphisms were associated with RA in the male population. The progression of radiographic severity, which was defined by a modified Sharp score plotted against disease duration, was significantly faster in the carrier of T allele at the -509 (p=0.048). CONCLUSION: Our data support the hypothesis that TGF-beta1 polymorphism may determine the progression of joint destruction in RA. PMID- 15794198 TI - beta2-adrenergic receptor gene single-nucleotide polymorphisms are associated with rheumatoid arthritis in northern Sweden. AB - The beta2-adrenergic receptor (beta2-AR) belongs to the group of G-protein coupled receptors and is present on skeletal and cardiac muscle cells and on lymphocytes. The gene encoding beta2-AR (ADRB2) displays a moderate degree of heterogeneity in the human population and the distributions of single-nucleotide polymorphisms (SNPs) at amino acid positions 16, 27, and 164 are changed in asthma, obesity, and hypertension and in the autoimmune disease myasthenia gravis. An involvement of the beta2-AR has also been suggested in human rheumatoid arthritis (RA) and its animal model. We describe here an increased prevalence of the alleles Arg16 and Gln27 and a lower prevalence of homozygosis for Gly16 and Glu27 in patients with RA. Patients having the genotype combination GlyGly16-GlnGlu27 had higher levels of rheumatoid factor (RF) and a more active disease than other patients. Patients having the genotype Arg16-Gln27+ had higher levels of RF when compared to those having Arg16+Gln27+, and patients who were carriers of Gln27 had a more active disease than non-carriers of Gln27. Our results show an association of beta2-AR SNPs with RA in a population from the northern part of Sweden. Our study also confirms the strong linkage disequilibrium of genotypes at amino acid positions 16 and 27. PMID- 15794199 TI - Health care costs attributable to the treatment of rheumatoid arthritis. AB - OBJECTIVE: A 'programme budget' for the resources used in the treatment and care of patients with rheumatoid arthritis (RA) was developed with a view of helping decision-makers assess the appropriateness of the current use of resources and to discuss future resource allocation. METHOD: The programme budget was developed using data from several national administrative registers. Patients with RA were identified by hospital diagnostic codes. The incremental cost of treating RA was defined as the difference in resource use for patients with and without RA. Incremental mortality due to RA was defined in similar way. Cost data were estimated for 5-year age groups. RESULTS: Patients with RA used on average 3.2 times as many health care resources as people without RA. The average 1997 incremental costs of primary and hospital care were EUR 253 and EUR 2.660 per patient respectively, corresponding to a national incremental cost of EUR 30 million (2000 price level). RA resulted in an annual loss of 1,549 life years. CONCLUSION: The programme budget approach is a useful tool in resource allocation decision-making, but discussions of alternative resource allocations must be based on robust studies of effectiveness and cost-effectiveness in the treatment of patients with RA. PMID- 15794200 TI - Marital and sexual satisfaction in patients with RA and their spouses. AB - This study sets out to determine predictors of marital and sexual satisfaction in patients with rheumatoid arthritis (RA) and their spouses. Fifty-nine patients and their spouses completed questionnaires independent from each other. Multiple correlations with marital and sexual satisfaction were computed for demographic variables, disease status, psychological distress, and social support. The results indicate that psychological distress and social support are more important than objectively assessed disease status in determining marital and sexual satisfaction in patients with RA. PMID- 15794201 TI - Apoptosis: the link between autoantibodies and leuko-/lymphocytopenia in patients with lupus erythematosus. AB - OBJECTIVE: To analyse the relation between serum autoantibodies and leuko /lymphocytopenia in patients with lupus erythematosus (LE). METHODS: Laboratory routine analyses (white blood cell counts, autoantibody detection), and flow cytometry (annexin V, CD3, CD4, CD8, CD95, F(ab)2 anti-human IgG) have been performed in LE-patients versus healthy controls. In vitro, the influence of pooled serum containing anti-dsDNA antibodies has been analysed on the CD95 expression. RESULTS: Leukocytes, lymphocytes, CD3+, CD3+ CD4+, and CD3+ CD8+ cells from LE-patients were significantly reduced compared with controls. Patients with autoantibodies had significantly lower absolute cell counts than those without. Apoptosis was increased in LE versus controls (p < 0.04). The percentage of CD95+ T-cells was increased, and the absolute number of CD95+ cells was reduced in LE. In vitro up-regulation of CD95 could be detected on T-cells of healthy donors. Induction of CD95 seems to be donor-dependent. CONCLUSION: The data suggest that autoantibodies may be associated with blood cytopenias. CD95 seems to play a central role in this signalling cascade. The underlying mechanism is unclear, but seems to be autoantibody-related apoptosis induction. PMID- 15794202 TI - A study of the genetic basis of C4A protein deficiency. Detection of C4A gene deletion by long-range PCR and its associated haplotypes. AB - OBJECTIVE: To study the frequency of a C4A gene deletions as the genetic basis of C4A protein deficiency (C4AQ0) and its associated haplotypes in Icelandic families with systemic lupus erythematosus (SLE). METHODS: Nine multiplex SLE families were genotyped for C4A gene deletions using a long-range polymerase chain reaction (LR-PCR) method, and major histocompatibility complex (MHC) haplotypes were defined. RESULTS: Of the SLE patients, first-degree and second degree relatives, 53.8%, 47.9%, and 28.6% had C4AQ0, respectively. A C4A gene deletion was found to be the genetic basis for C4AQ0 in 64.3% of SLE patients, 60.0% of first-degree and 50.0% of second-degree relatives. All individuals carrying haplotype B8-C4AQ0-C4B1-DR3 had a deletion, and the deletion was also found on haplotypes B8-C4AQ0-C4B1-DR7 and B7-C4AQ0-C4B1-DR3. CONCLUSION: The study shows that a C4A gene deletion is the most common genetic basis for C4AQ0. It accounts for two-thirds of C4AQ0 and is found on different MHC haplotypes. One third of C4AQ0 is due to other as yet undefined genetic changes. The results demonstrate a heterogeneous genetic background for C4AQ0, giving further support for the hypothesis that C4AQ0 may be an independent risk factor for SLE. PMID- 15794203 TI - Biopsy-verified response of severe lupus nephritis to treatment with rituximab (anti-CD20 monoclonal antibody) plus cyclophosphamide after biopsy-documented failure to respond to cyclophosphamide alone. AB - BACKGROUND: The monoclonal anti-B cell antibody rituximab (Rituxin, Mabthera) may be of benefit in antibody-driven diseases, including systemic lupus erythematosus (SLE) nephritis. PATIENTS AND TREATMENT: Two female patients with biopsy confirmed severe and active SLE nephritis despite treatment with cyclophosphamide (CyX) were given four rituximab infusions plus two additional CyX infusions. RESULTS: Both patients tolerated the treatment well and SLE activity improved. On repeat kidney biopsy after the combined treatment, Patient 1 showed a profound reduction of nephritis activity, and she was maintained on low-dose prednisolone only. A repeat biopsy after 1 year confirmed the sustained reduction of lupus nephritis activity. In Patient 2, rebiopsy after combined treatment also showed a significant reduction in disease activity. CONCLUSION: These cases provide histopathological documentation of a significant treatment benefit from rituximab plus CyX in two patients refractory to CyX alone. This combination is being explored further as salvage therapy for such CyX-resistant patients. PMID- 15794205 TI - Elevated IgG4 concentrations in serum of patients with Mikulicz's disease. AB - Mikulicz's disease has recently been included within primary Sjogren's syndrome. It is a unique condition involving enlargement of the lacrimal and salivary glands, characterized by few autoimmune reactions. It is responsive to glucocorticoid treatment. Analysis of IgG fractions was performed in patients with Mikulicz's disease in order to determine the differences between Mikulicz's disease and Sjogren's syndrome. The study showed that serum IgG4 concentrations are elevated in patients with Mikulicz's disease, but not in those with Sjogren's syndrome. PMID- 15794204 TI - Insulin-like growth factors I and II in the aseptic loosening of total hip implants. AB - Periprosthetic interface tissue and pseudocapsule samples surrounding aseptically loosened hip implants and control knee synovium were studied by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. Endothelial cells, fibroblasts, and monocyte/macrophages contained bone formation enhancing insulin-like growth factors (IGFs). In interface tissue we found fewer IGF-I and IGF-II positive cells than in control tissue. In pseudocapsular tissue we found fewer IGF-I positive cells and an equal amount of IGF-II positive cells compared to control tissues. Decreased bone formation may contribute to net loss of bone around aseptically loosened hip implants. PMID- 15794206 TI - Kikuchi-Fujimoto disease associated with mixed connective tissue disease. AB - Kikuchi-Fujimoto disease is characterized by painful cervical lymphadenopathy and constitutional symptoms. Microscopical study of lymph nodes shows focal areas of non-suppurative necrosis with histiocytic and plasmacytoid cell infiltrates. The course is usually benign. Often primitive, necrotising histiocytic lymphadenopathy may be associated with autoimmune disorders. We describe the case of a 30-year-old female patient with two 15-day courses of Kikuchi-Fujimoto disease flares within a period of 3 months, occurring in association with mixed connective tissue disease. PMID- 15794207 TI - Bronchiolitis obliterans organizing pneumonia in a patient with Behcet's disease. AB - A 55-year-old male presented a productive cough, haemoptysis, and intermittent fever for 2 weeks. He was diagnosed as having Behcet's disease (BD) with manifestations of recurrent oral ulcers, genital ulcers, erythema nodosum, a gastro/duodenal ulcer, colon ulcers, and folliculitis. After admission, a chest X ray showed bilateral ill-defined patches. Subsequent chest computerized tomography revealed multiple patchy consolidations over the left and right lower lobes. Because of poor response to oxacillin antibiotic treatment, an open lung biopsy was arranged. Bronchiolitis obliterans organizing pneumonia (BOOP) was diagnosed. Methylprednisolone pulse therapy, followed by prednisolone and cyclophosphamide maintenance, was prescribed. The patient responded well, with clinical and radiological resolution. Early diagnosis with proper treatment is crucial in the management of this lethal pulmonary disease. PMID- 15794208 TI - The antiphospholipid/cofactor syndrome: results of routine screening for antibodies to beta2-GPI upon suspicion of the antiphospholipid syndrome. PMID- 15794210 TI - The lucky ones. PMID- 15794209 TI - Acute pseudogout in a patient with treated acromegaly. PMID- 15794211 TI - Three roads back: whats fair got to do with it? PMID- 15794212 TI - Three roads back: scars on a woman. PMID- 15794213 TI - Three roads back: shot between the eyes. PMID- 15794214 TI - Resetting the brain. PMID- 15794215 TI - Aspirin anomaly. PMID- 15794216 TI - Unwelcome houseguests. As the IRS expands CEP audits to include smaller healthcare facilities, Carle Foundation Hospital finds itself playing host. AB - It's not your ordinary audit. If the Internal Revenue Service decides to give your healthcare organization a Coordinated Examination Program audit, be prepared for a long haul-two to three years. And it's hard to anticipate if you'll be targeted. Catherine Jacobson, left, at Rush University Medical Center, says the IRS "told us there was no reason why they chose us, except we were on their list because of our size". PMID- 15794217 TI - Sympathetic ears. Legislators agree with acute-care hospitals' stance. PMID- 15794218 TI - Budget woes. Medicaid, Medicare likely targets. PMID- 15794219 TI - Defending CPOE. Advocates downplay study attributing errors to IT. PMID- 15794220 TI - Billing dispute, ACORN stages walk-in at AHA headquarters. PMID- 15794221 TI - Little help in bankruptcy bill. PMID- 15794222 TI - Tenet's prospects. Physicians may hold key to turnaround. PMID- 15794223 TI - Pushing for worker insurance. PMID- 15794224 TI - DaVita gets another subpoena. PMID- 15794225 TI - CNA wins latest staff-ratio battle. PMID- 15794226 TI - Health Care Hall of Fame. Lifetime of leadership. Scott Parker led Intermountain's rise to top of nation's integrated systems. PMID- 15794227 TI - Health Care Hall of Fame. Promoting ethics, equality. Stuart Wesbury Jr. honed leadership skills in a variety of healthcare settings. PMID- 15794228 TI - Let the spin begin. Greater transparency urged for device trials. PMID- 15794229 TI - Code blues. Healthcare succumbs to a national trend toward unethical conduct. PMID- 15794230 TI - Lifelong learning. To stay ahead of the trends, continue your education throughout your career. PMID- 15794231 TI - The doctor in the dell. Wisconsin center's specialty: diagnosing and treating illnesses most likely to crop up in agricultural professions, rural populations. PMID- 15794232 TI - Using questionnaire and cotinine concentration in urine for studying the smoking habits and ETS exposure of pregnant women. AB - The aim of this study was to evaluate the exposure of pregnant women to tobacco smoke on the basis of a standardised survey questionnaire, and determination of cotinine in urine. In the study participated 133 women delivering, checked in at the Obstetric Department of Independent Public Health Care Unit in Srem in 2003. The all women fill in the questionnaire forms referring smoking and exposure to ETS (Environmental Tobacco Smoke). In the tested group 22.6% were smoking during pregnancy. Only 5 pregnant women quit smoking, however around 60% of smokers reduced the number of cigarettes. Near 56% of women were exposed to ETS, mainly during social events. In case of women, who declared smoking the mean concentration of cotinine was 2164 ng/mg of creatinine. Passive smokers had cotinine concentration between 10 and 50 ng/mg of creatinine Cotinine may be used to validate tobacco smoke exposure assessment from surveys. PMID- 15794233 TI - [Smoking mothers and symmetry of growth in newborns born in 2000 in the Pomeranian region]. AB - In many publications concerning norms of weight in adults and child population the appropriate body weight is defined in connection to its stature. PI initiated in 1970 by Miller seems to be of a great value. Relative risk of morbidity and mortality in full term newborns with low value of nutrition index increases from 1.9 to 4.2 times. The aim of this paper was evaluation of symmetry of intrauterine development based on PI calculation. The particular remarks was paid to newborns from mothers smoking during the time of pregnancy. 10770 children born in 2000 in 8 hospitals of Pomeranian Region and 2119 newborns from 1990 born in Institute of Obstetrics and Gynecology Medical University of Gdansk were analyzed. The estimation of frequency of smoking during pregnancy was randomized on questionnaire done in the group of 2627 mothers from 1990, 1999, 2002, 2004. The mean value of PI in the group of full term newborns from 19990, was 2.19 +/- 0.27. In 2000 this value was 2.05 +/- 0.25 and was significantly lower than previous. Newborns from smoking mothers had significantly lower PI than those from women who did not smoke during pregnancy. Estimating harmonious of intrauterine growth in the group of newborns from smoking mothers I stated the greater lack of body mass than its length, so it is more asymmetric. It is similar to pathology which act on fetus in the 3rd trimester of the pregnancy. Now the newborns are slimmer as mean PI shows. I notice a disproportionate intrauterine growth in the cohort of newborns from smoking mothers. In this group the estimation of PI is advisable. PMID- 15794234 TI - [Tobacco smoking and body weight of newborns]. AB - Literature data about the effects of tobacco smoke constituents in pregnancy are reviewed. Analysis on the authors material comprises 73 smoking and 70 non smoking women in pregnancy. In the former group the natal body weight of new born is by 308 g lower and the placental-fetal index by 0.02 higher with no differences in the body weight increase of smoking and non-smoking women in pregnancy. PMID- 15794235 TI - [Cigarette smoking and the age of natural menopause in women in Poland]. AB - The empirical basis for this work has been the material obtained from cross sectional studies carried out in the years 1998 to 2002, among women living in Poland. The material included data on 3793 women aged from 35 to 69 years. The women who participated in the research varied with regards to the age, education and socio-economic status. Menopausal status and median age at menopause was estimated with the use of retrospective and status-quo methods. It found out that the median age at natural menopause, estimated by probit analysis, was 50.33 years. The main aim of this work is to define whether and to what extent does smoking of cigarettes influence the age of natural menopause. Smoking of cigarettes increases the risk of earlier menopause (value of test F=138.8, p<0.0001). Non-smoking women experience menopause on average two years later than women who are habitual smokers. Also the number of cigarettes smoked every day differentiated the age of menopause, accelerating its occurrence in women who smoked more than five cigarettes per day (value of test F=50.81, p<0.0001). PMID- 15794236 TI - [The socio-economic status of women smoking during pregnancy and birth weight of their newborns]. AB - In this survey, socio-economic status of pregnant women smoking during pregnancy and influence of this status on birth weight of their newborn were assessed. The research was done on the group of 1328 pregnant women. Smoking habit was declared by 18.6% of women and passive exposure to tobacco smoke was stated by 30.6% of respondents. Significant association between active and passive smoking and socioeconomic status of respondents was observed. Cigarette smoking during pregnancy was the main cause of lower birth weight of newborns. The newborns of actively smoking mothers were lighter of about 357 g according to those ones of mothers who did not smoke and were not exposed to passive smoking, and were lighter than newborns of mothers who were not exposed to passive smoking of about 330 g. The passive exposure to the tobacco smoke was not significant considering effect on the birth weight. PMID- 15794237 TI - [The awareness of health consequences of active and passive smoking and access to information about these consequences in the population of pregnant women]. AB - Despite of significant progress in the area of perinatal care, there have been numerous perinatal complications still existing. The health status of mother and newborn is affected by many factors, namely: socio-economical; medical prior to pregnancy and medical through the pregnancy; dietary customs, substance use, including tobacco smoking; health service quality; and risk factors that may occur through fertilization. The active and passive exposure to tobacco smoke in utero have been linked to numerous gestation and delivery complications, including low birth weight, preterm delivery and low Apgar score. In this survey, the awareness of health consequences of active and passive smoking in the population of pregnant women and access to the information of this consequence was assessed. Among of women smoking through their pregnancies, the only 19.7% got the information from their doctors about harmful effect of active and passive smoking, and 16.5% of respondents declared self-awareness on health effects of exposure to the tobacco smoke. PMID- 15794238 TI - [Active and passive exposure to tobacco smoke of pregnant women: two-center study]. AB - According to the current data over 12 million Polish citizens smoke tobacco, 40% of them are women. Maternal smoking in pregnancy significantly increases the rate of having a low-birthweight baby, the rate of perinatal mortality and several other adverse pregnancy outcomes. The aim of the study was to estimate the rate of smoking women during pregnancy and those who are exposed to secondhand smoke. 219 women hospitalized at Gynecology and Obstetrics Department of Wroclaw Medical University and Gynecology and Obstetrics Department in Opole have undergone the examination. The pregnant women have been interviewed and the results of questionnaire have been checked by the exhaled carbon monoxide (ExhCO) measurement. CO expresses the percentage of carboxyhemoglobin (HbCO) and is used for objective monitoring tobacco smoking. The results of the study suggest that regular exposure to secondhand smoke becomes a serious problem. 25.5% patients from Wroclaw and 16.4% patients from Opole have declared passive smoking. The mean value of ExhCO in this group of women obtained the level of 3.3 ppm vs 1.95 ppm in the group of patients who have denied both active and passive exposure. PMID- 15794239 TI - [Comparative study of pregnant women exposure to tobacco smoke: POLAND-MEXICO]. AB - The aim of the study was to estimate both active and passive pregnant women exposure to tobacco smoke. 154 women hospitalized at the Gynaecology and Obstetrics Department in Opole, Poland and at the Gynaecology and Obstetrics Department of Hospital Universitario Monterey in Mexico have undergone the examination. The patients have been surveyed in order to obtain a subjective evaluation of their exposure. Then the exhaled carbon monoxide (ExhCO) measurement used for objective monitoring tobacco smoking has been performed. According to the results of the questionnaire 40 women (26% of the examined patients) confirmed smoking in pregnancy. 24 women (60% of smoking ones) have been smoking for the whole period of pregnancy. Though the rate of smoking among Polish and Mexican women was comparable, Polish mothers were less likely to quit after confirmation of pregnancy. Both Polish and Mexican smoking mothers were much younger than their non-smoking counterparts. The babies of the smoking mothers have been noticed to obtain the lower count in Apgar scale. The exhaled carbon monoxide (ExhCO) measurement has revealed that 14 Polish and 10 Mexican patients have been exposed to tobacco smoke. The results of their questionnaires have been thoroughly examined. 71% of them confirmed active smoking in pregnancy, 8.3% pointed to second-hand smoke exposure, 20.7% lied by denying the exposure. Thus the need to verify the questionnaires results has been confirmed. The mean value of ExhCO of both non-smoking women and those, who quit smoking after confirmation of pregnancy was equal. This may suggest that quitting smoking in the early stages of pregnancy reduce the risk of the harmful smoke impact on the unborn baby. Nearly doubled average concentration of ExhCO among passive smoking mothers in comparison to the non-smoking ones arises a serious concern. PMID- 15794240 TI - [Evaluation of the women's knowledge about Fetal Tobacco Syndrome]. AB - Smoking is still a common habit in Poland. Nowadays, it is the way of coping with stress, it is used to become calm or more alert. It is also widely accepted by people from various backgrounds. Unfortunately, the number of young women- tobacco addicts is increasing. Smoking before and during pregnancy has bad effects on women and their children as well. First discoveries of negative influence of smoking observed lesser birth weight of the babies whose mothers were smokers, compared with the babies whose mothers were non-smokers (Simpson 1957). He proved in his research that the more the mother smokes the lesser the birth weight of her baby is, and it doesn't depend on the woman's age, weight, height or gynecological past. Pregnant women who smoke more often suffer from such consequences as: bleeding during pregnancy, placenta abruption, placenta previa, premature rupture of membranes. The fetus death rate after the twentieth week of pregnancy is much higher among smoking women than non-smoking ones. Lower fertility and conception difficulties are also consequences of smoking. Therefore, the education of girls and women in the period before they plan having children is very important. The mother and child's health depends exclusively on the knowledge, awareness and attitudes of future parents. PMID- 15794241 TI - [The sperm quality in fertile smokers]. AB - In this prospective study different semen parameters of fertile smokers were compared with fertile non-smokers. 58 men with proven fertility (17 smokers and 41 non-smokers) at the same age were evaluated. Smokers had significantly less spermatozoa with motility grade A (p<0.01), B (p<0.05), and C (p<0.05), and more abnormal forms in the ejaculate (p< 0.001). The percentage of sperm with head defects (p<0.05) and cytoplasmic droplets (p<0.001) were also significantly higher in smokers. No differences were found in the assessment of semen volume, total sperm count, hypoosmotic swelling test, induced acrosomal reaction and acidic aniline blue staining test between two groups (p>0.05). However the concentration of leukocytes in semen was significantly higher in smokers (p<0.001), which can suggests oxidative imbalance in the ejaculates of these men. Our data demonstrate that cigarette smoking is associated with reduced semen quality which could worsen fertilizing potential in fertile men. PMID- 15794242 TI - [Influence of cigarette smoking on some sperm parameters in males with decreased fertility]. AB - 112 patients from infertile couples with diagnosed asthenospermia and oligoasthenospermia have been examined. The results have been analysed in view of smoking. An increased number of antisperm antibodies have been detected in smoking patients. No difference between the two groups examined according to WHO standards have been found. PMID- 15794243 TI - [Symptoms of andropausal syndrome in smoking and non-smoking males]. AB - Symptoms of andropause syndrome such as: erectile dysfunction, somatovegetative and psychic symptoms have been examined in groups of smoking and non-smoking patients between 45 and 75 years of age Tests of testosterone, prolactin and SHBG levels have been carried out. Earlier andropause, a lower level of testosterone as well as more common arterial hypertension have been found in the group of smoking patients. PMID- 15794244 TI - [Perioperative anxiety in female smokers subjected to gynaecological surgery--a preliminary report]. AB - Anxiety constitutes one of the most important psychological factors influencing body reaction to surgical injury; in gynecological patients anxiety ratings appear especially high. Cigarette smoking belongs to essential risk factors in the perioperative period and may result in increased levels of anxiety. A pilot study has been carried out with the aim of evaluating the impact of smoking habit on the perioperative anxiety level and postoperative pain in women undergoing gynecological surgical procedures. In 9 smokers and 10 non-smokers, anxiety was assessed by means of the State Anxiety Score of the Spielberger State-Trait Anxiety Inventory (STAI) and the visual analog scale (VAS), preoperatively and on the second day after surgery. Additionally, intraoperative and postoperative opioid analgesic uses were analyzed and on the second day after surgery patients rated their recall of postoperative pain with the use of the VAS. Postoperative State anxiety was lower than that reported preoperatively only in non-smokers (median: 53.0-40.0, p=0.02, Wilcoxon p=0.02). Smoking did not significantly influence intra- and postoperative opioid analgesic requirements and postoperative pain. CONCLUSIONS: This study suggests that in female smokers subjected to gynaecological surgery cigarette smoking may contribute to altered anxiety kinetics in the perioperative period. PMID- 15794245 TI - [Cigarette smoking practices in patients with malignant neoplasms of the oral cavity and jaws]. AB - The aim of this study was to estimate the cigarette smoking practices in patients with malignant neoplasms of the oral cavity and jaws, who were treated in the Outpatient Department of the Maxillofacial Department of Silesian Medical Academy in Katowice from january 2000 to june 2004. It was found that significant number of patients (100% of men and 72.72% of women) with oral squamous cell carcinoma had been smoking in the period of time preceding appearance of neoplasm. The men smoked more cigarettes a day and started smoking in a younger age than women. Gender factor did not influence significantly the period of time from starting of smoking practices to the appearance of neoplasm. The patients with the other types of neoplasms denied cigarette smoking. PMID- 15794246 TI - [The role of environmental factors in HPV infections of the upper respiratory tract of healthy children]. AB - INTRODUCTION: Human papillomavirus (HPV) has been universally recognized as the etiologic factor of many malignant and non-malignant diseases of the upper respiratory tract, skin and female genital tracts. Up to date more than 100 various genotypes of HPV have been recognized. The process of the HPV transmission and the risk factors of spreading the infection have been not confirmed explicitly. THE AIM OF THE STUDY: The aim of the paper was the identification of environmental factors of spreading infection in the material of asymptomatic carriers of HPV. MATERIAL AND METHODS: A group of 201 pre-school children (109 girls and 92 boys) was laryngologically examined after getting a consent from their parents. The virological examination using PCR methods reveal 58 asymptomatic carriers of HPV and 143 HPV-free children. RESULTS: The analysis of history and data reveal significant relationship between the HPV infection and the number of active smokers in the family (38.2% vs 19.4%). The dental caries was found more often in HPV positive children. The fact worth consideration is the standard of living, which in the families of HPV infected children was estimated frequently as low or rather low. CONCLUSIONS: Exposure to unfavorable environmental factors like active smokers in the family and low standard of living can led to spreading of the HPV infections. PMID- 15794247 TI - [Comparative analysis of nicotine dependence and motivation to quit smoking in patients with bronchial asthma or COPD]. AB - The aim of the study was to assess differences in tobacco addiction rate and motivation to quit tobacco smoking in patients with bronchial asthma and chronic obstructive pulmonary disease (COPD). The study comprised 384 males from Warsaw and its environs, aiming to quit smoking within the nearest year, without cardiovascular or neoplastic diseases, with at least medium education (17.19). The subjects were qualified into one of three groups: a control group--subjects without pulmonary diseases (n=143), patients with COPD (n=126) and patients with bronchial asthma (n=115). The smoking index (pack-years) was calculated in each group. The tobacco addiction rate was assessed with the Fagestrom questionnaire, and the motivation to quit smoking with the Schneider test. All data were obtained during individual examinations. The mean smoking index in the study population was 29.0 +/- 20.6 pack-years (control--27.0 +/- 20.6; asthma--25.2 +/- 14.2 and COPD--38.0 +/- 21.4). A high tobacco addiction rate was found in 40.5% of the patients with COPD, 37.1% of the patients with asthma and 30.3% of the subjects from the control group. The differences between the groups were not statistically significant (p>0.05). In the asthmatic group the percentage of subjects with a high motivation to quit smoking was significantly lower (48%) than in the COPD group (74%, p=0.005) and the control group (75%, p=0.002). It was found that patients with COPD significantly more frequently than asthmatics and people without pulmonary diseases need medical assistance in overcoming nicotine addiction. PMID- 15794248 TI - [Active tobacco smoking as determinant of costs of inpatient treatment of COPD exacerbations]. AB - The aim of the study was assessment of the influence of tobacco smoking on direct costs of in-hospital treatment of chronic obstructive pulmonary disease exacerbations (COPD). 112 men who underwent in-hospital treatment of exacerbation of moderate COPD in the years 2001-2002 were included in the study. The patients were qualified into one of three groups: I--never-smoking patients (n=40); II- former smokers (n=42); III--active smokers (n=30). The mean age of patients was 70.9 +/- 8.9 years (I--74.8 +/- 7.4; II--68.4 +/- 9.4; III--69.4 +/- 9.8). The mean direct cost of COPD patient treatment was PLN 2162.00 +/- 920.90 (I--PLN 2025.50 +/- 681.30; II--PLN 1875.80 +/- 608.70; III--PLN 2744.60 +/- 1266.20). The mean period of smoking was 18.3 pack-years in the group II and 27.7 pack years in the group III. No significant differences in FEV1 were found between the groups. No significant differences were found in the treatment costs between the groups I and II. It was demonstrated that the costs of in-hospital COPD treatment are significantly higher in the group III than in the groups I and II. A linear regression formula was calculated, showing a linear relationship between each consecutive pack-year of addiction and the costs of exacerbation treatment in the group III (R=0.39, R2=0.13, standardized BETA index = 24.0; p<0.03). No similar relationship was found in the former smokers' group. The analysis of the data shows that: (1) active tobacco smoking by patients is related to significantly higher direct costs of in-hospital treatment of COPD exacerbations than in case of never-smoking patients or patients who smoked in the past, and (2) the costs of in-hospital treatment of COPD exacerbations in case of people who have stopped smoking are comparable to the costs of hospitalization of never-smoking patients with COPD. PMID- 15794249 TI - [Prevalence of chronic obstructive pulmonary disease (COPD) and other pulmonary diseases among participants of the program of early detection and prevention of COPD]. AB - Chronic obstructive pulmonary disease (COPD) is one of the most frequent chronic diseases and the sixth leading cause of death in the world. The cigarette smoking is the most important cause of the disease and therefore, that COPD is preventable. Programmes detecting COPD in its preclinical phase using spirometry with successful smoking cessation prevent disease progression. The aim of the study was to assess the prevalence of COPD and other pulmonary diseases among participants of the program of early detection and prevention of COPD. The program was held in 2002 and carried out by eleven health care institutions. Active smokers over 40 insured in the Fund were offered free spirometry, chest x ray, individual and group behavioural therapy to stop smoking, and chest physician's examination for patients with diagnosed pathology. The results confirm high detection of COPD and other diseases and confirm the efficacy of targeted screening for early detection of COPD and other diseases. PMID- 15794250 TI - [Passive smoking, atopy and respiratory health in schoolchildren]. AB - Aim of the study was to verify the hypothesis that the effect of passive smoking exposure on the occurrence of respiratory symptoms and diseases is higher in children with increased individual susceptibility defined on the basis of occurrence of symptoms of atopy. Project was performed as the cross-sectional epidemiological study which comprised population of 1,534 children attending primary schools in Swietochlowice, Kedzierzyn-Kozle, Pszczyna and Kielce. On the basis of standard questionnaire an information regarding occurrence of wheezing, attacks of dyspnea with wheezing, asthma and spastic bronchitis was collected. Atopy in child was defined on the basis of occurrence of symptoms of allergy to at least one of the following allergens: house dust mites, animal dander, pollens or symptoms of food allergy confirmed by physician's diagnosis. Exposure to tobacco smoke was assessed also on the basis of the information regarding smoking given by the parents in the questionnaire. The effect of interaction atopy * passive smoking was assessed on the basis of variable called "interaction" which had three levels: level A: when exposure to passive smoking and atopy = 0, level B: when exposure to passive smoking or atopy =1 and level C: when exposure to passive smoking and atopy =1. The results of our study revealed that statistically significant relationship between interaction atopy * passive smoking on the occurrence of analyses symptoms and diseases. The occurrence of symptom/disease increased with the presence of either atopy or exposure to passive smoking and was the highest when atopy and exposure to passive smoking was present. The results of simple analyses were conformed by multivariate analyses. The results of our study suggest that the effect of environmental exposures could be higher in children with atopy. PMID- 15794251 TI - [Effect of passive cigarette smoking on the frequency of respiratory tract infections, allergy and bronchial asthma in children]. AB - Passive cigarette smoking is an important medical problem; it concerns first of all children as an injurious environmental factor affecting their health. Results of the conducted questionnaire and evaluation of the CO concentration in exhaled air proved the injurious exposure to be the cigarette smoke. The study showed more often allergy, bronchial asthma and respiratory tract infections in the group of children exposed to passive smoking cigarettes at home in comparison with the group of children without this exposure. The study was conducted among 75 families belonging to Model Family Practice so it has also important educational aspects. PMID- 15794252 TI - [Exposure to tobacco smoke among three years old children in Bialystok and its influence on occurrence of respiratory tract diseases]. AB - Addiction to tobacco smoking is very popular among young and middle age people in Poland, who are parents of pre-school children as well. Respiratory tract diseases are the main reason of visits of pre-school children in General Practice surgery. Anonymous questionnaires were given to the random sample of mothers of three years old children. 311 respondents were chosen from whole population of 1200 children attending kindergartens in Bialystok. The questionnaires showed that half of the examined children are involuntary exposed to tobacco smoke at home 20% also outside home (neighbors, homes and, cafeterias, cars etc). We did not find any relationship between exposure to smoke and occurrence of respiratory tract infection in children. We presume that this may be apparently the result of the fact that many smoking parents try to not to smoke in the presence of their children. Increasing awareness of harm resulting from passive smoking is the benefit of educational campaigns in Poland. PMID- 15794253 TI - [Effect of smoking on phonation]. AB - The aim of this paper was presentation of voice estimation in smokers, who did not complain of voice disorders. The examination was performed in 20 patients with history of chronic tobacco using (at least 20 years). The videolaryngostroboscopy and acoustic voice analysis were performed. The specific changes as in organic dysphonia were found in larynx in both examinations. The most important finding in acoustic voice analysis was abnormal parameter describing vFo--Fundamental Frequency Variation and lower comparing to normal voices fundamental frequency. PMID- 15794254 TI - [Effect of tobacco smoking on amylase activity in patients with pancreatitis]. AB - The pancreas is one of the first organs pathologically affected by the tobacco smoking. However, the mechanism of development of these changes is not eventually recognised. It has been demonstrated that nicotine influences exogenous function of pancreas. The aim of this study is to prove the influence of tobacco smoking on amylase activity in serum and urine of smoking and non-smoking patients with diagnosed acute (AP), chronic (CP) and chronic exaggerated pancreatitis (CEP). Serum and urine has been collected from 57 patients with AP, CP and CEP. The activity of enzyme has been determined using the colorimetric method with ethylidene-G7-PNP (4,6-ethylidene-p-nitrophenyl-alpha,D-malthoheptozyde). The nicotine metabolites has been assayed with the immunoenzymatic method (ELISA) using rabbit polyclonal antibodies against cotinine. The highest amylase activity in serum and urine has been observed in smoking patients with CEP. Much higher differences in amylase activity has been estimated in urine of patients with CP and AP (CP: in non-smoking patients more than three times lower activity than in smoking patients; AP: in non-smoking patients more than two times lower activity than in smoking patients). It has been revealed that the differences in amylase activity in both serum and urine in smoking patients in comparison with non smoking patients with pancreatitis may prove a significant influence of tobacco smoking on exocrine function of pancreas. PMID- 15794256 TI - [Smoking among thyroid patients]. AB - The effects of cigarette smoking on the thyroid disease (Graves disease, toxic nodular goiter, toxic adenoma) have been studied for years. However, the effect of smoking on thyroid function is still controversial. OBJECTIVES: To determine the impact of cigarette smoking on the development of above-mentioned thyroid disease. PATIENTS METHODS: Study of 405 individuals with different thyroid disease. Information on thyroid disease and smoking habits was gathered by Fagerstrom questionnaire, and the patients' endocrinologist or general practitioner verified the diagnosis. RESULTS: The effect of smoking was more pronounced in Graves' patients than in other thyroid patients. We found out significant number of patients with toxic nodular goiter. CONCLUSIONS: Smoking is associated with an developing clinically thyroid disease--especially Graves disease. We also suggest the connection between psychological traits and smoking in this group of patients, corresponding with our previous examinations. PMID- 15794257 TI - [High level of hsCRP in patients with Graves' ophthalmopathy--preliminary study]. AB - C-Reactive Protein (CRP) levels are used to indicate the presence of an inflammatory process that may be caused by bacterial infection or pathological processes. The objective of this study was to compare the levels of CRP and hsCRP (high sensitivity CRP) in smoking and non smoking patients with Graves' Ophthalmopathy (GO). Study included 59 hyperthyroid patients (age > 18 years, 50 females, 9 males, mean +/- SD: age 34 +/- 16) with varying degrees of severity of GO (classification according to NOSPECS) with hyperthyroidism prepared to radioiodine therapy. Smoking status was determined by Fagerstrom test and Brinkman's index. CONCLUSIONS: 1. Smoking additionally stimulates inflammatory processes in the patients with Graves' Ophthalmopathy, 2. Smoking patients with PMID- 15794255 TI - [The effect of smoking on serum nitric oxide (NO) in acutely carbon monoxide poisoned patients]. AB - An important role of Nitric Oxide (NO) in relaxing vascular smooth muscle is commonly known. In this pilot study we aimed to asses an impact of cigarette smoking on NO serum concentration in acutely CO poisoned patients. The measured marker of cigarette smoking was urea cotinine. A highest NO concentration and lowest diastolic blood pressure were noted in the subgroup of acutely CO poisoned smokers. Diastolic blood pressure in the subgroup of not CO poisoned non-smokers and was higher compared to acutely CO poisoned (both smokers and nonsmokers) and also to subgroup of those person who were exposed only to CO in cigarette smoke. PMID- 15794258 TI - [Vitamin A levels among nonsmoking mothers of children with orofacial clefts married to a smoker]. AB - Passive smokers have lower plasma levels of pro-vitamin A (beta-carotene) than do unexposed nonsmokers. Maternal insufficiency of vitamin A (retinol) during pregnancy results in congenital abnormalities in the offspring. Our aim was to examine vitamin A concentrations in passive smoking and non-smoking mothers of children with isolated orofacial clefts. SUBJECTS AND METHODS: Plasma samples from 68 never smoking mothers and 15 non-smoking mothers married to a smoker (> or = 20 cigarettes/day) were analyzed for retinol. All participants answered a questionnaire about socio-demographic characteristics and lifestyle behaviors. Retinol concentrations were measured by high pressure liquid chromatography (HPLC). RESULTS: Passive smokers have significantly lower plasma retinol concentrations (1.86 micromol/L vs. 2.21 micromol/L; p<0.05) and ratio of plasma retinol to plasma total cholesterol levels (0.33 micromol/mmol vs. 0.48 micromol/mmol; p<0.05) than never smokers. Women married to a smoker had also lower socioeconomic status. CONCLUSIONS: Polish nonsmoking mothers of children with orofacial clefts married to a heavy smoker are probably at increased risk to have low plasma retinol levels. Usefulness of higher periconceptional supplementation with vitamin A in that subset of women requires further research. PMID- 15794259 TI - [The normotensive carriers of the MTHFR 677T allele, displaying the increased risk of development of the abdominal aortic aneurysm (AAA), occur at the highest frequency among the smoking patients]. AB - Abdominal aortic aneurysm (AAA) presents itself as a progressive dilation of the abdominal aorta, leading--if untreated--to rupture. It is a common disease of the elderly, with a complex etiology. Smoking, hypertension and several genetic factors are recognized as relevant for the pathogenesis of AAA. We studied association between the polymorphism of the MTHFR (methylenetetrahydrofolate reductase) gene within the fourth exon (677C>T) and the occurrence of hypertension and smoking status in the group of 74 male patients with AAA. In the patients group, the smoking hypertensive persons represented the largest subgroup (43%). We determined the the MTHFR 677C>T polymorphism in AAA patients and compared it to that in 71 healthy normotensive males. The frequencies of the 677T allele and MTHFR 677C>T genotypes were similar in both groups, but the subgroup of normotensive AAA patients (n=29) displayed significantly increased frequencies of 677T allele (0.4) and of 677CT and TT genotypes (69%), as compared to those in the control group (0.28 and 46%, respectively). This corresponds to the 3.3-fold greater risk of AAA in normotensive subjects with the 677T allele of MTHFR, as compared to the homo-zygotes 677CC (p<0.03; 95% CI=1.2-9.2). The highest frequencies of MTHFR 677T allele (0.43) and 677CT and TT genotypes (73%) were found in the subgroup of normotensive smoking patients (n=22). PMID- 15794260 TI - [The level of cotinine in urine and placenta of women smoking or exposure to tobacco smoke during pregnancy]. AB - The aim of the studies was evaluation smoking habit by delivering woman using objective method--determination of cotinine in urine and placenta. In the study participated 93 woman delivering in Gynaecological-Obstetric Clinical Hospital of Medical University at Warsaw. Evaluation of tobacco smoking and exposure to ETS performed on the base of questionnaire and level of cotinine in urine. In the group of non smoking woman cotinine was not detected any in urine any in placenta. In the second group (19 patients), woman exposure to ETS concentration of cotinine in urine was 28.9 +/- 14.9 ng/mg of creatinine and only in two placentas was detected a low level of cotinine. In smoking delivering woman the urine concentration was 837.0 +/- 1324.5 ng/mg of creatinine and 21.3 +/- 7.5 ng/g placenta calculated from all results and 67.0 +/- 3.5 ng/g calculated only from positive results. A weak relation between concentration of cotinine in placenta and urine of smoking woman was shown. PMID- 15794261 TI - [Evaluation of the tobacco smoke exposure among students living in students' dormitories]. AB - Passive smoking results from involuntary smoke inhalation in the environment where tobacco and other tobacco products are smoked. This is a common situation in numerous public objects, as restaurants, offices, dormitories and students' clubs. Non-smokers are exposed to very high concentrations of tobacco smoke in these places. Environmental exposure to tobacco smoke of the inhabitants of the Pharmacy School dormitory in Sosnowiec was studied. The evaluation was based on urine cotinine concentration measured with the HPLC method. Additionally, a Fagerstrom test was conducted among declared smokers in order to evaluate the degree of their pharmacologic addiction to nicotine. The obtained results indicate that non-smoking dormitory inhabitants are strongly exposed to environmental tobacco smoke, but active smokers did not presented with pharmacological addiction to nicotine. PMID- 15794262 TI - [Experimental assessment of exposure to the tobacco smoke and its effect on superoxide dismutase and glutathione peroxidase activities]. AB - Activities of two antioxidant enzymes, superoxide dismutase (SOD) and glutathione peroxidase (GPx) were determined in plasma of rats exposed to tobacco smoke (500, 1000 and 1500 mg CO/m3 air in 5 days) and to 1000 mg CO/m3 in 1, 2 and 3 weeks. When compared to controls, the SOD activity was decreased in the exposed animals, the lowest smoke dose gave the most pronounced effect and the highest dose revealed the least diminution. GPx activity was statistically higher in the rats inhaled with the highest smoke dose. Time of exposure did not effect GPx but SOD raised in correlation to week number, although it did not reach control values over the experiment period. The observed alterations in plasma probably mirror the organ adaptation to oxidative stress in animals exposed to tobacco smoke. PMID- 15794263 TI - [Total radical trapping antioxidants parameters on plasma in smoking and non smoking matched maternal-cord pairs]. AB - Oxidative stress in smoking pregnant women is assumed to be enhanced by oxidants and free radicals of tobacco smoke. Therefore we studied the effect of cigarette smoking during pregnancy on plasma total radical trapping antioxidants parameters (TRAP) in mothers and their babies. Pregnant women were selected into the groups according to concentration of cotinine in serum. TRAP was determined by method of Wayner et al. using oxygen electrode (YSI Model 5331). The plasma concentration of TRAP was lower in smoking pregnant women than in tobacco abstinent group (533.0 micromol/l vs 600.0 micromol/l in I trimester, 578.0 micromol/l vs 583.5 micromol/l in II trimester and 583.5 micromol/l vs 589.0 micromol/l in III trimester. The differences were statistically significant in first and second trimester (p<0.05). Value of TRAP in umbilical cord blood from newborns of smoking mothers was significantly lower and amounted only 74% of that observed in non-smoking ones (366.5 micromol/l vs 492.0 micromol/l; p<0.001). Plasma level of TRAP was significantly correlated with serum concentration of cotinine both in smoking pregnant women (r=-0.32; p<0.01) and in umbilical cord blood of their children (r=-0.49; p<0.01). Our results indicate that smoking depletes plasma TRAP in mothers and their babies. Decreased level of total plasma antioxidants parameters may have a negative effect on antioxidant protection systems in neonates. PMID- 15794265 TI - [Determination of cadmium in urine of tobacco smoking pregnant women]. AB - Tobacco smoke contain few thousands of chemical compounds, among them heavy metals. From toxicological point of view most important are lead, cadmium and radioactive polonium 210. The aim of the study was determination of cadmium in urine of tobacco smoking pregnant woman and checking if there is a correlation between the concentration of cadmium and cotinine, the most frequently used tobacco smoke biomarker. The study showed that concentration of cotinine in urine of smoking women was 702.5 +/- 1131.4 ng/mg creatinine and ranged from 50 to more than 6000 ng/mg creatinine. Cadmium concentration in smokers was 1.6 +/- 2.6 ng/ml and ranged from 0 to 11.5 ng/ml. In urine of woman who do not smoke and are not exposure to ETS was 1.1 +/- 2.2 ng/ml in the range 0-2.5 ng/ml and was not statistically different from concentration of cadmium in urine of smoking pregnant woman. In any one non-smoking woman, concentration of cadmium was not higher than 5 ng/ml, but in 11.8% of smoking women this level was exceeded. Calculations showed a weak correlation between concentration of cadmium and cotinine in urine of smoking pregnant women. PMID- 15794264 TI - [Tobacco smoke effects the activity of superoxide dismutase, glutathione peroxidase and total antioxidant status in pregnant and non-pregnant animals]. AB - Total antioxidant status was measured as the reduction of the ABTS radical cation as well as the activities of SOD and GPx in female rats exposed and non-exposed to the cigarette smoke, pregnant and non-pregnant. The assessment was done in lungs, plasma, kidneys, liver and placenta of Wistar rats exposed to the cigarette smoke (1500 mg CO/m3 air) for 21 days. Total antioxidant status was significantly elevated in lungs and plasma of smoke-exposed animals, pregnant and non-pregnant, when compared to the matched controls. In other examined tissues antioxidant capacity was diminished in all tested groups of animals. Activities of SOD and GPx were markedly decreased in tissues of all examined animals. Exposition to the cigarette smoke, despite some changes due to the adaptation to stress, diminishes the antioxidant capacities of the body and in the pregnant animals it may effect the pregnancy outcome. PMID- 15794266 TI - [The influence of smoking and passive smoking to occurrence of metals in breast milk]. AB - Concentrations of 7 metals (Cd, Cr, Cu, Fe, Mn, Pb and Zn) have been assayed in the breast milk of three groups of women, inhabitant in Cieszyn region. The first group (n=34) comprised the women non-smoking, the second (n=29) passive smoking women, and third (n=18)--women smoking and exposed to cigarette smoke. The concentrations of all the trace metals were found to be higher in the breast milk of women smoking and passive smoking. The results obtained were analyzed using the principal component analysis (PCA). PMID- 15794267 TI - [Effect of tobacco smoke exposure on urine lead concentration of pregnant women]. AB - Among other toxic compounds, tobacco smoke contains also heavy metals, for example lead. Lead can cross the placenta barrier and have negative influences on fetus development. In the study compared urine lead concentration of pregnancy women exposed to tobacco smoke (active smokers and exposed to environmental tobacco smoke--ETS) with unexposed. There was find that mean urine lead concentrations in groups of pregnant women exposed and unexposed to tobacco smoke was no different (F2,189=0.2 p>0.8). However it was found that in urine of women exposed to tobacco smoke the concentration of lead increase with increase the concentration of cotinine (r2=0.14, t106=4.2, p<<0.0001). PMID- 15794268 TI - [Cadmium level in the human saphenous vein used in coronary artery bypass grafting]. AB - The human saphenous vein is the main superficial vein of the lower limb. It begins on the foot, runs across among medial surface of shin and knee and frontal medial surface of thigh. Saphenous vein remains the most widely used bypass conduit for the treatment of occlusive coronary and peripheral vascular disease. In vivo it exhibits rhythmic contractile activity. The cellular mechanisms controlling saphenous vein activity are poorly understood. It is likely that saphenous vein contraction is preceded by electrical activity followed by an increase in intracellular calcium. The conducted research has shown that environmental exposure to cadmium leads to an accumulation of these metals in the organism and the disturbance in the metabolism of such important elements as calcium. The objective of the present research was to describe the concentration of cadmium in human saphenous vein in vitro by atomic absorption spectrometry. An association between cigarette smoking and concentration of cadmium is reviewed. PMID- 15794269 TI - [Lead and cadmium occurrence in deciduous teeth of children exposed to cigarette smoke in apartments]. AB - The aim of this study was to find out if children's exposure to tobacco smoke in the apartments influences the lead and cadmium content and the values of Pb/Ca and Cd/Ca ratios in hard tissues of their deciduous teeth. Material consisted of 71 shed deciduous teeth; 37 from children in the age of 6 to 13 years additionally exposed to tobacco smoke in the apartments and 34 from children in the same age, whose apartments were free of tobacco smoke. Lead, cadmium and calcium content was determined by means of flame atomic absorption spectrometry. Deciduous teeth of children additionally exposed to tobacco smoke in the apartments exhibited higher lead and cadmium content and higher values of Pb/Ca and Cd/Ca ratios, which indicated significant accumulation of these metals, when compared to deciduous teeth of children whose apartments were free of tobacco smoke. PMID- 15794270 TI - [Analysis of the possible interaction between cadmium and nickel in the gallstones from actively and passively smoking and non-smoking women]. AB - The problem of statistical characteristics of occurrence cadmium and nickel content in gallstones from active and passive smoking and no smoking women living in southern Poland is presented in the work. The subject of the research were gallstones, gained intraoperatively from 93 women (24 active smoking, 18 passive smoking, 51 no smoking women). The content of cadmium and nickel was determined using atomic emission spectroscopy with inductive couple plasma (ICP-AES). The influence of smoking to women describes the degree of elimination cadmium and nickel from women organism by use of gallstones in detoxication process. PMID- 15794271 TI - [Application of PCA (Principal Components Analysis) for the estimation of smoking effect on the occurrence of trace elements in women's gallstones]. AB - The PCA (Principal Components Analysis) was used to estimation of the role of smoking in the changes of elements contents in gallstones. The concentration of given elements were determined by ICP-AES method. It was stated that smoking regardless of sex, influence on the occurrence of elements in hydroxyapatites of gallstones, in particular it concerns changes of As, Pb, Zn, Se, Ti contents. PCA method let to describe the role of selected elements in entire chemical composition of gallstones coming from smoking women. PMID- 15794272 TI - [Analysis of possible interaction between lead and calcium in gallstones from active and passive smoking and no smoking women]. AB - The problem of interaction between lead and calcium in gallstones from active and passive smoking and no smoking women living in southern Poland is presented in the work. The subject of the research were gallstones, gained intraoperatively from 93 women (24 active smoking, 18 passive smoking, 51 no smoking women). The content of lead and calcium was determined using atomic emission spectroscopy with inductive couple plasma (ICP-AES). The place of living, smoking and age play an important role in changing the lead and calcium content of gallstones. PMID- 15794273 TI - [Different distribution of some metals causing intoxication of teeth in smoking and non-smoking people]. AB - The purpose was analysis in the individual species of teeth cigarette smoking and non-smoking people. Also relationships between concentration of some metals in environment and teeth men and women living on seven given towns Silesian Province (Jaworzno, Bedzin, Dabrowa Gornicza, Tychy, Czeladz, Sosnowiec, Katowice). PMID- 15794274 TI - [Metal content in the femur head of non-smokers and ex-smokers]. AB - The purpose of this work was the elemental analysis of 12 heavy metals concentration in some part in femur capitulum smoking and non smoking women and men, living in Silesian Towns: Katowice, Siemianowice, Czeladz. Femoral capitulum were obtained at total hip replacement operations of 86 women and men with idiopathic osteroarthrosis which smoking in early time. PMID- 15794275 TI - [Arsenic in hydroxyapatites in gallstones from actively and passively smoking and non-smoking women]. AB - The investigation on the arsenic changes carried out in gallstones from 93 women (24 active smoking, 18 passive smoking, 51 no smoking women). The elemental composition of gallstones was determined by ICP-AES method. For smoking women in comparison to men the changes of given elements were different. Negative role of smoking in particular concerned accumulation of Mg, Si, P, As. PMID- 15794276 TI - [The significance of social support in the maintenance of smoking abstinence in the population smoking cessation program]. AB - According to I.G. Sarason social support is the help provided to the individual in difficult and stressful situations. In this study we have assumed that quitting smoking is a very stressful situation for the smokers. Therefore, they need social support from the others (family, friends, co-workers, health personnel). DATA SOURCES: 1. One-year follow up survey of a random sample (n=993) of the Quit & Win 2000 participants in Poland. The data collection method was obtained through mail questionnaire (response rate = 65.6%). 2. Results of the comparative data analysis of one-year follow up survey from 20 countries organizers of the Quit & Win 2000. The aims of the study were to recognize: 1. impact of social support on the quitting smoking process from the decision to participate in the campaign till the maintenance of one-year abstinence, 2. lack of social support as the reason for relapse. RESULTS AND DISCUSSION: In Poland the Quit & Win 2000 participants achieved the highest continuous complete abstinence rate (43%, mean=26.3) and the lowest social support (no support 42%, mean= 25.9) out of 20 countries. Analysis of data suggested that the social support was the important factor for smokers in encouraging them to participate in the campaign and try to quit. There aren't statistical relationships between social support and one-year abstinence (p>0.05) as well as between lack of support and relapse (p>0.05). CONCLUSIONS: The significance of social support in quitting smoking process needs the further exploration. PMID- 15794277 TI - [Attitudes and behaviors related to smoking habit among elementary and high school students]. AB - Survey in Krakow (2001) preceding implementing of the anti-nicotine program in elementary schools and gymnasiums was done in order to determine the frequency of smoking, attitudes towards smoking habit and smokers and the influence of family on developing these attitudes. In this survey questionnaire consisting of 32-41 questions (depending on the school type) on socio-demographic data and attitudes scale was used. Statistical analyses were done using chi2 test to evaluate significant relations between variables and multilevel logistic regression models to establish risk. Obtained results show, that in the recent years the number of pupils initiating smoking is not diminishing (13.8% of gymnasium pupils and 3.0% of elementary school pupils smoke and 19.3% and 8.2% smoked in the past, respectively). In majority pupils' attitude towards smoking was negative (89.2% in elementary schools and 74.5% in gymnasiums). However, attitudes towards smokers were more differentiated. Lack of defined attitude towards smokers presented 62.6% of elementary school pupils and only 32.4% of gymnasium pupils. Although smoking is not approved by majority of pupils, in particular gymnasium pupils are more tolerant towards smokers and more often have positive attitude towards smokers (25.8%) in comparison with their younger colleagues. The fact of lack of determined attitude towards smokers among 42.1% of gymnasium pupils and 30.3% of elementary school pupils is distressing. Odds ratio of lack of determined attitude towards smokers increases when at least one of parents smokes (OR=2.05) and also when older colleagues smoke and peers do not smoke (OR=24.5). It decreases for those, who were characterized by lack of acceptance for smoking (OR=0.172). Obtained results confirm the role of social environment (especially family) in developing of attitudes towards smokers among pupils. They also the presence of pupils together with psychosocial immaturity characteristic for process of development favors ambivalence of judgments and beliefs connected with this habit. PMID- 15794278 TI - [Tobacco smoking among high school adolescents]. AB - According to the American data each day more than 4000 kids in the United States try their first cigarette and more than 2000 kids become new regular, daily smokers. Roughly one-third of all youth smokers will die prematurely from smoking caused disease. The aim of the study was to estimate the frequency of nicotine addiction among high school adolescents. The special attention has been paid to the problem of passive smoking. 60 high school students from Opole and Zielona Gora have undergone the examination. The exhaled carbon monoxide (ExhCO) measurement has been performed. Moreover the students have been interviewed about the frequency of smoking and smoking habits in their surroundings. In 17 cases (28% of students) the values of the exhaled carbon monoxide (ExhCO) concentration were out of range for non-smoking people. The exhaled carbon monoxide (ExhCO) measurement has revealed that 65% of smoking children should be regarded as heavy smokers, proving their strong nicotine addiction. 21 children (35% of examined population) have confirmed exposure to secondhand smoke. In the families of 36 students (60%) at least one of the parents has been smoking cigarettes. PMID- 15794280 TI - [Some elements of life style, including cigarette smoking, among adolescents]. AB - The aim of the study was to determine some of elements of life style among 336 pupils attending randomly chosen high school (in 4 provinces) were studied by the questionnaire. 12.5% of adolescents were smoking cigarettes. Physical activity of half of respondents was ranked as average, 1/3 was underweight, 3.9% was over weight--(obesity--0.9%). Most of adolescents ate 3 meals a day, 92% were snacking. The main nutritional faults were small frequency of consumption of milk and milk products, vegetables and fruits, fish and vegetable oils. PMID- 15794279 TI - [Active and passive smoking among students of Polish high schools]. AB - The survey was conducted among 29540 students in different secondary schools. All interviewers were 17.42 +/- 1.14 years old. The aim od this work was to define the relation between smoking adolescent and their health. The group of smokers consisted of 5112 (60.26%) men and 3370 (33.70%) women. The main causes of initiating smoking reported by respondents included curiosity--(47.91%) followed by stress (39.82%) and alcohol (6.77%). Smoking tobacco by parents at home results in inspiring by their children the unwholesome ingredients of tobacco smoke. Frequent staying among tobacco smokers significantly influences decision of starting or giving up smoking tobacco (59.24%). The obtain results may be helpful in preparing effective educational and preventive project among students, which can eliminate or lower the influence of factors predisposing to start and continue smoking tobacco. Special programs should be worked out for children showing early health consequences of smoking cigarettes. Giving up smoking is possible for "young" smokers, if they have not developed the symptoms of addiction. PMID- 15794281 TI - [Potential risk of the consumption of non-prescription drugs in an elderly hypertensive subject on nicotine substitute therapy--case presentation]. AB - In this paper the case of 68-year-old female who consumed the drug to aid smoking cessation is presented. This drug is taken by her simultaneously with 16 other (including 7 prescribed drugs and 9 non-prescribed). According to the list of her medicines it may be concluded that she suffers from hypertension (which is also declared by her) and heart failure or atrial fibrillation. The additional treatment with the drug to aid smoking cessation (Tabex) may increase the blood pressure. The control of blood pressure may be even more difficult if non prescribed drugs with hypertensive effect are taken. In analyzed subject there were two nonsteroidal antiinflammatory drugs (Ibuprom, Voltaren acti) and ginseng extract (Bodymax). In summary, it must be underlined that in elderly subject on nicotine-substitute therapy uncontrolled consumption of over the counter drugs may potentially worsen the result of hypotensive therapy. PMID- 15794282 TI - [Effect of tobacco smoking on taste sensitivity in adults]. AB - This study aimed to evaluate the influence of the tobacco smoking on taste sensitivity in healthy adults. The study involved healthy subjects, aged 30 to 60 years. The experimental group consisted of 28 smoking subjects; the control group consisted of 33 non smokers. The ability to recognize four basic types of taste and thresholds for detection of sweet and salty taste were determined in this study. The measurement of taste thresholds were carried out by electrogustometry. The obtained results have been found higher thresholds for detection of salty taste in smokers compared with control subjects (p<0.05). We estimated that such tastes as: sweet, salty, sour have been recognized by smokers weaker and their sensitivity of tongue receptors to electric stimulus was slightly lowered than nonsmokers. PMID- 15794283 TI - [Application of the principles of minimum intervention by primary health care doctors--evaluation by patients addicted to nicotine]. AB - The aim of the study was to analyze to what extend the principles of minimum intervention are applied by basic health care doctors in Mazovia. The study comprised 326 tobacco smokers, divided into two groups: group I--including 160 adults living in towns with a population below 10000 and group II--including 166 inhabitants of towns with a population above 100000. Each subject had their tobacco dependence rate (the Fagerstrom test) and the period of smoking (pack years) calculated. Each subject evaluated application of the minimum intervention principles by their basic health care doctor anonymously. The data obtained were statistically analyzed. The study groups did not differ in terms of nicotine addiction rate. It was found that almost half of the patients smoking tobacco had never been asked by their doctor about smoking habit during consultation. An educational material concerning the harmfulness of smoking is received by every seventh smoker, and an attempt of a pharmacological therapy of addiction is undertaken only in every tenth patient. The minimum medical intervention principles in tobacco smokers are twice more often applied by basic health care doctors working in small towns. PMID- 15794284 TI - [Determining the possibility of collecting reliable data for use in decision making in health care on the example of cost-effectiveness analysis of methods used in smoking cessation]. AB - Reports on Polish Health care system found that the evidence from systematic reviews and cost-effectiveness analyses is not sufficiently used in decision making. The aim of this study was to determine the possibility of reliable cost and other data collection in Poland to inform cost-effectiveness analysis of smoking cessation methods in Poland, which could be used to support decisions from the perspective of National Health Fund. In order to estimate long term effects of smoking cessation Markov model was built. Direct medical costs of the following smoking related diseases were sought: chronic obstructive pulmonary disease, coronary heart disease, lung cancer and stroke. Analysis was performed from National Health Fund perspective. Published cost data were insufficient for the analysis, so an attempt was made to obtain cost data from National Insurance Fund, which was not very successful due to their lack. Mortality data were obtained from National Office of Statistics. Lung cancer incidence and mortality were obtained from national register. No registries systematically and routinely collecting incidence and prevalence data for other diseases were localized. No studies exploring utilities of Polish population in the analyzed health states were found. As no disease registries, except for cancer, systematically collecting epidemiologic data and no unified system of collecting cost data were found, less reliable data has to be used in cost-effectiveness analyses supporting the decisions in health care in Poland. PMID- 15794285 TI - [Tobacco use by career soldiers of the Polish Armed Forces--the start of a breakthrough?]. AB - Cigarette smoking in the Polish Armed Forces has a long standing tradition. Various organizational processes which operated for many years encouraged the development of cigarette smoking through various means, including allocating supplies of cigarettes or funds for their purchase. Smoking the Polish Army continues at a high rate and depending on the specific unit may total from 40% to 65%. This results from the lack of any anti-smoking campaigns as well as a nearly total disregard for anti-smoking smoking regulations. A recent development in the armed forces is a reduction in the number of smokers among the younger professional corps as well as 'contract' soldiers. Surveys conducted among these groups have indicated a greater concern for health related issues and sports activity. As has become apparent, the examples viewed in other armies of NATO have had a significant impact, as the number of smokers in those forces amounts to no more than 10-30%. It is clear from the surveys that among factors which have contributed to the decision to cease smoking are the fact that physical conditioning has increased in importance as a prerequisite for candidates for professional service. Despite the promising changes among younger professional soldiers, the problem of a high percentage of smokers among the older (age-wise) and conscripted soldiers remains as an issue of concern. PMID- 15794286 TI - [Use of rough sets theory in analysis of qualitative data in toxicology studies]. AB - In analysis of medical data a problem of simplified and not effective analysis of qualitative data is frequently encountered. This situation takes place particularly when data are measured in the weakest measuring scale--nominal scale. However this scale contains less detailed information than interval or ordinal scale, there exist a lot of different mathematical approaches to examine existing dependencies, among other causality. In this paper we present the exam pie of usage of technique based on theory of rough sets to toxicological data gathered during mediumistic program "Throw smoking together with us" in 2000. The obtained results were compared with results estimated with alternative techniques published in [1]. PMID- 15794287 TI - [Nicotine replacement therapy--method for smoking cessation]. AB - Nicotine replacement therapy (NRT) play a major role in pharmacotherapy of nicotine dependence. Currently various nicotine formulations are sold as over-the counter (OTC) products in several European countries. The effectiveness and safety of nicotine formulation was proved in many countries in population studies. The most often used NRT products are nicotine chewing gum and Nicotine transdermal patch. PMID- 15794288 TI - [Molecular and cellular changes following exposure to tobacco smoke causing laryngeal cancer. An outline of the problem]. AB - The incidence of laryngeal cancer is strongly connected with exposure to tobacco smoke containing at least 40 genotoxic carcinogens. DNA lesions induced by tobacco smoke carcinogens can be turned into stable mutations. Oncogenes and tumor suppressor genes are first of all responsible for initiation of carcinogenesis. The same time cells provide such self-protection processes as: carcinogen detoxication, DNA repair and apoptosis. The initiation of carcinogenesis followed by oncogenesis results from the competition between DNA lesions/mutations formation and the protection processes; the latter are genetically controlled. PMID- 15794289 TI - [Safety of Zyban administration in nicotine dependence treatment]. AB - Zyban (bupropion hydrochloride, produced by Glaxo Wellcome Group) is a drug available in Poland used in a nicotine dependence treatment, as an aid in smoking cessation. The aim of the study was to review available information concerning observed side-effects of Zyban. The information provided by producer, articles and reports published in MEDLINE and EMBASE databases since January 2001 till September 2004 were analysed. Side-effects were analysed, compared and classified according to anatomy-physiological systems, where these side-effects are observed. PMID- 15794290 TI - [Preparing pharmacists for participating in health promotion programs as exemplified by the smoking prevention program]. AB - Health promotion is an interdisciplinary subject which glue together such sciences as medicine, psychology, sociology, social politics and many others. It appears to be the most interesting theme for the pharmacists. One of the most important problem carried in the drug-stores is the fight against the nicotine dependency, which gives the pharmacists a large field to show his knowledge and competence in helping people with the smoking problem. To make the pharmacist patient cooperation happen, we need to instruct chemists at pre-, and postgradual courses about treating the nicotine addiction. We cannot underestimate the pharmacists' role in the whole fighting nicotine dependency process. Modern pharmacist is fully qualified to give tips or guide patient who get to finish his addiction. PMID- 15794291 TI - [Tobacco and health in the teaching of nature and biology]. AB - The author discusses instruction on the harmfulness of smoking tobacco in the Polish educational system. This concerns primarily two subjects, Nature and Biology, in which this issue appears. In this context, he examined the content of their syllabi, manuals, and teacher's guides. He presents the most common didactic solutions employed, and comes to the conclusion that this field lacks original studies and novel proposals. On his part, he puts forward some suggestions concerning the kind and scope of measures that can be taken at various educational levels. He also stresses the need for parallel and continuous social and administrative action. PMID- 15794292 TI - 'Culture clues' help staff understand their diverse patients and families. PMID- 15794294 TI - CMs and interpreters start DP at admission. PMID- 15794293 TI - End-of-life issues may be most difficult for DP staff. PMID- 15794295 TI - Is your falls prevention program getting results? PMID- 15794296 TI - Create a paper trail to improve teaching, communication. PMID- 15794297 TI - Patient safety alert. Keystone project yields results in six months. PMID- 15794298 TI - Attracting diverse patients is part of a strategic plan. PMID- 15794300 TI - Is it redundant to have DPs in addition to CMs? PMID- 15794299 TI - Discharge planning advisor--the update for improving continuity of care. New system streamlines patient throughput. PMID- 15794301 TI - [Dialysis....at what cost?]. PMID- 15794302 TI - [Diagnosis of renal cysts]. AB - Evaluation and management of renal cysts Renal cystic diseases are a heterogeneous group of conditions including heritable, developmental, and acquired disorders. They are united by the presence of microscopic or giant fluid filled cavities and affect both children and adults. The definitive diagnosis of many of the renal cystic diseases requires clinical, radiological, pathological, and genetic analysis. A precise diagnosis is essential for prognosis, treatment, and future genetic counselling. PMID- 15794303 TI - [Selective COX-2 inhibitors and cardiovascular risk]. AB - Selective cyclooxygenase-2 (COX-2) inhibitors were initially developed to reduce the gastrointestinal toxicity of conventional NSAIDs. However, it became rapidly evident that selective COX-2 inhibitors may increase the risk of cardiovascular events. Today, the mechanisms for the potentially adverse cardiovascular effects of coxibs remain unknown, and it is increasingly clear that our understanding of the role of COX-2 in cardiovascular function is incomplete. Based on some available indirect evidence, and unless more clear-cut data become available, the longterm use of high doses of selective COX-2 inhibitors, in elderly patients or patients with a high cardiovascular risk should be limited whenever possible. PMID- 15794304 TI - [IgA nephropathy: what you have need to know in 2005]. AB - Although considered as a benign glomerulopathy, IgA nephropathy (IgAN) is now a well-known cause of end-stage renal disease (ESRD). Fifty percent of people suffering from IgAN developp renal insufficiency and 20 to 30% may reach ESRD after 20 to 25 years of evolution. ACEI is indicated to obtain a thigh control of blood pressure and to reduce proteinuria. Corticosteroids alone or in association with immunosuppressants are indicated for agressive, proliferative form of the disease or when there is an unfavorable outcome despite symptomatic treatment. PMID- 15794306 TI - [Prevention of contrast nephropathy: myth or reality?]. AB - Prevention of Contrast Media induced acute renal failure is of importance, especially in patients with chronic renal failure. Vasodilatators such as fenoldopam and prophylactic hemodialysis have been reported to procure disappointing results. Hydratation prior to and post- exposure to contrast media remains the cornerstone of prevention. Urine alcalinisation with sodium bicarbonate may bring additional renoprotection. Use of a small quantity of contrast media and Acetylcysteine concomitantly to hydratation are also recommended. PMID- 15794305 TI - [Urine electrolytes: use and pitfalls to avoid]. AB - In clinical situations of electrolytic or acid-base disorders, the measure of electrolytes concentration and osmolality in a urine sample, which is called the urinary spot, is a simple way to guide the diagnosis and therapy. The interpretation of results must take into account the patient's history and clinical examination. The urine sodium concentration and urine osmolality are key elements in the diagnosis of dysnatremias and in renal failure. The urine chloride concentration is useful in the diagnosis of acid-base disorders. The potassium excretion in the urine, which is regulated mainly by aldosterone, is particularly helpful in the case of hypokaliemia. Finally, the determination of the transtubular potassium gradient is a valuable tool for the diagnosis of hypoaldosteronism. PMID- 15794308 TI - [Treatment of lung cancer: critical review of new agents]. PMID- 15794307 TI - [BMP-7 (Bone morphogenetic protein-7): a future treatment for chronic renal failure?]. AB - Renal fibrosis is an evolutive process frequently leading to end stage renal failure. Several studies have demonstrated the dominant and mutually antagonistic roles of two proteins from the transforming growth factor superfamily. BMP-7 has a protective effect on the kidney, by decreasing apoptosis, maintaining and restoring the epithelial phenotype and dysplaying anti-fibrotic activity. The mechanisms through which TGF beta 1 favours fibrosis are still unclear, although most of its activity appears to be due to antagonism of the effects of BMP-7. Animal studies are very encouraging, as exogenous BMP-7 has allowed stabilisation and even regression of renal fibrosis with a concomitant stabilisation or improvement of renal function. The therapeutic implications of these findings are promising. PMID- 15794309 TI - [Treatment of cancer pain]. PMID- 15794311 TI - [Bioethics, the great European uproar]. PMID- 15794312 TI - [New methods in bone and joint imaging]. PMID- 15794310 TI - [Face transplantation: psychological, social, ethical and legal considerations]. PMID- 15794313 TI - [War and medicine]. PMID- 15794314 TI - Turning our collective attention to Medicaid. PMID- 15794315 TI - Childhood obesity: what can be done to help today's youth? AB - Childhood obesity is at epidemic proportions in the United States today. This epidemic has created a public health crisis. Although several initiatives are under way to address childhood obesity, including legislative bills before Congress and a call by the National Academy of Sciences for society to band together to stop the rise in the rates of childhood obesity, none of these efforts address what can be done today to help those youth that are considered morbidly obese and who have failed to find solutions with the standard medical treatment plan. This article discusses the use of bariatric surgery as a possible solution, presents a case study to illustrate the impact bariatric surgery can have on youth who are morbidly obese, and defines the pediatric nurse's role in helping to assure that youth are included in the current legislative process. PMID- 15794316 TI - Caring for the child who is obese: mobility, caregiver safety, environmental accommodation, and legal concerns. AB - Two thirds of Americans are considered overweight, while 25% are obese. Obesity is the most common health problem facing U.S. children today, regardless of ethnicity, gender, or socioeconomic status. Lack of preplanning for the hospitalized obese child threatens both patient and caregiver safety, and is becoming a serious consideration in pediatric nursing. Some of the challenges in mobilizing larger children with an eye to preventing caregiver injury and promoting patient safety are explored. Equipment, policy formation, education, and legal considerations are outlined within this context. A case study is presented herein. PMID- 15794317 TI - Creation of a bariatric surgery program for adolescents at a major teaching hospital. AB - Due to the alarming increase in adolescent obesity, pediatric surgeons and children's hospitals are exploring the use of bariatric surgery as a treatment option in a highly selected group of children. PMID- 15794318 TI - Cyclic vomiting syndrome: a disease in disguise. AB - Cyclic vomiting syndrome (CVS) is a childhood disorder characterized by bouts of vomiting that last from a few hours to several days. The current prevalence in the pediatric population is estimated at 2% (Li & Balint, 2000). The vomiting episodes occur in a regular cycle: some children have them every few days, others every few months. Currently, researchers have been unable to determine what causes CVS. Many of the patients and families, however, can identify particular events that seem to trigger the vomiting. Diagnosis can be difficult because CVS does not leave clues that can be found by any current diagnostic test. Lack of knowledge about CVS by physicians, nurses, and the lay public contribute to the helplessness families feel when coping with this disease in disguise. Nursing's primary actions in the care of children and their families affected by CVS should be one of advocacy for diagnosis and support for the family and child through treatment and long term management of this chronic illness. PMID- 15794319 TI - Voices for America's children: an interview with Deborah Stein. Interview by Veronica D Feeg. PMID- 15794320 TI - Teens: rural, obese and poor: lessons we learned in Wal-mart. PMID- 15794321 TI - Leukotrienes: their role in the treatment of asthma and seasonal allergic rhinitis. PMID- 15794322 TI - Indoor Air Quality: part II--what it does. AB - Newton, MA. A recent report indicated air quality samples taken from several rooms in the town's North High School had elevated CO2 levels of 2,000 parts per million (ppm) (Viser, 2004). State standards set 800 ppm as the optimum reading. Although not an immediate health issue, high CO2 levels are indicative of poor air circulation--clean air comes in but stale air is not vented out. Safety issues arise in the school setting when chemicals or toxic substances are in use and cannot be vented, therefore posing the health risk (Viser, 2004). Poor Indoor Air Quality (IAQ) in schools can result in decreased academic performance and days lost due to illness in the school age population (Environmental Protection Agency [EPA], 2003). As the school nurse at North High School, what would you do? PMID- 15794323 TI - What happened to the rhythm? PMID- 15794324 TI - Toddler with a chronic cough. PMID- 15794325 TI - Rapid critical appraisal of randomized controlled trials (RCTs): an essential skill for evidence-based practice (EBP). PMID- 15794326 TI - Kangaroo holding beyond the NICU. AB - The primary goal for maternal-child nurses in caring for parents and their infants is to support parental confidence and competence through education and services to assure optimal parental-infant attachment. Kangaroo holding in the hospital and home environment is one intervention that encourages parental-infant interaction as well as provides the benefits of closeness, proximity, and touch. Nurses in hospitals, private practice, the community, and beyond must work together to change practice by adding the intervention of kangaroo care for the benefit of infants and their families. PMID- 15794327 TI - One small boy. PMID- 15794328 TI - Letter regarding "arsenic in a child's world". PMID- 15794329 TI - Regarding "antibiotic sampling" article. PMID- 15794330 TI - Building confident organizations by filling buckets, building infrastructures, and shining the flashlight. AB - Leaders have the ability to create confidence or fear in the organization. Kanter (2004) writes that confidence is the "...sweet spot between arrogance and despair" (p. 8). Overconfidence causes people to overshoot, and to assume they are invulnerable. Under-confidence is just as harmful because it leads to under investing in people, under-innovating, and eventually leads to the disenfranchisement of staff and poor morale. If we take Kanter's advice, we will build an infrastructure that creates confidence in everyone and the organization. But we will not stop there. We will focus "flashlights" on people and activities that are inspiring to others. PMID- 15794331 TI - Application of structural descriptors for the evaluation of some physicochemical properties of selected bile acids. AB - Selected bile acids such as: cholic (C), glycocholic (GC), glycodeoxycholic (GDC), chenodeoxycholic (CDC), deoxycholic (DC), lithocholic (LC) and glycolithocholic acids (GLC) were separated by TLC technique. Selected topological indices, based on adjacency and distance matrices, and electrotopological states were calculated for the studied bile acids. Different possibilities of application of structural descriptors to calculate specified physicochemical data of the examined bile acids were found (QSPR). Usability of structural descriptors for the estimation of chromatographic separations of the examined bile acids was shown only for optimal separation conditions. Of all topological indices, only the index C allows to describe the order of adsorption of the examined bile acids (QSRR). Whereas, Gutman's index Mv was proved to be most useful for the estimation of lipophilicity of the examined bile acids (QSAR). The studies indicate that the range of applicability of given structural descriptors is limited for certain analytical and physicochemical problems. PMID- 15794332 TI - Kinetics of degradation of enalapril maleate in dosage forms. AB - The effect of humidity and temperature on the stability of enalapril maleate (ENA) in tablets have been studied. Tablets with ENA were stored in final packs (blisters) and without final packs in the atmosphere of increased humidity (76.4 % RH) and in the temperature range from 313 K to 333 K (forced degradation test). Degradation of ENA in tablets stored in final packs and without final packs followed the first order reaction model. During this study, the product of hydrolysis of the ester group and intramolecular cyclization have been observed. Thermodynamic parameters (activation energies, enthalpy and entropy) for the decomposition of ENA in tablets were calculated. PMID- 15794333 TI - Dental gel viscosity parameters and pharmaceutical availability of non-steroidal anti-inflammatory drugs. AB - Model prescription for dental anti-inflammatory gels with carboxymethylcellulose sodium salt and non-ionic surfactants have been worked out. Viscosity parameters of 10 variant gel forms were investigated and an attempt on their interpretation was undertaken in relation to pharmaceutical availability of non-steroidal anti inflammatory drugs (diclofenac, ibuprofen sodium). Viscosity tests demonstrated higher pharmaceutical availability of ibuprofen sodium than of diclofenac sodium particularly from model gels with surfactants of low number of oxyethylene segments in the structure. The above has been confirmed by in vitro studies on the kinetics of therapeutic agent penetration into external compartment. PMID- 15794334 TI - Investigations on prolonged release of vitamin C formulations. AB - In this study, investigations have been carried out to prepare prolonged release formulations of ascorbic acid in order to obtain a more effective dissolution profile of the drug. For the preparation of micropellets, the fluid bed method was chosen and to obtain prolonged effect, aqueous dispersions of Eudragit S, L and their mixtures were used. After setting the parameters of fluid bed, micropellets were produced using different coating dispersions. Following the production process, morphology, friability, particle size distribution, flow properties, bulk density and dissolution tests were carried out on the micropellets. The most appropriate formulation for our aim was chosen according to these tests. PMID- 15794335 TI - Investigation of some topical formulations containing dexpanthenol. AB - Owing to its ability to regenerate epidermal cells Dexpanthenol (D-panthenol; chemically known as (+)-2,4-dihydroxy-N-(3-hydroxypropyl)-3,3-dimethylbutyramide) has found use for the treatment of patholytic ileus and postoperative distention. The purpose of research was to develop a gel containing dexpanthenol by monitoring the effect of various concentrations of a gelating agent on the activity of the ciliary apparatus. A system containing 2.5% of hydroxyethylcellulose was optimal for the preparation of the gel. Together with a formulation containing 5% of dexpanthenol, drops with equal concentration of the active compound were tested for comparison. Physical characteristics, such as osmotic pressure, acidity, density and viscosity of the preparation were determined as well as its microbiological sterility. The anti-inflammatory activity of the gel was determined following its topical application. Epidermal tests showed its good tolerance after topical application to the shaved skin of guinea pigs. PMID- 15794336 TI - Effect of adriblastin on viability, cell cycle and apoptosis in B16 and cloudman s91 mouse melanoma cells in vitro. AB - Recently, the oncologists have been increasingly interested in apoptosis. which is morphologically. biochemically and topographically different from necrosis. Apoptosis is probably a key point to understand the resistance of malignant cells to cytotoxic drug. The aim of this study was to determine the influence of adriblastin on morphology and kinetics of cell proliferation and evaluation of apoptosis and necrosis in B16 and Cl S91 mouse melanoma cells in vitro. Adriblastin induced morphological changes indicating occurrence of apoptosis and necrosis in B16 and Cl S91 mouse melanoma cells. The number of viable B16 and Cl S91 cells was estimated by flow cytometry analysis. Apoptotic cells were detected using the annexin V-FITC test. DNA content, after staining cells with propidium iodide, was analysed by flow cytometry. B16 cell line was more sensitive to adriblastin treatment than the CIS91 cell line. The adriblastin EC50, value estimated for the B16 line was 2.4 microM. while for Cl S91 line was 4.0 microM. It was revealed that about 59% Cl S91 cells and 65% B16 cells had died in the apoptotic way after adriblastin addition at EC50, concentration to B16 and Cl S91 cultures. Adriblastin induced cell arrest in G1 and S cell cycle phase in both B16 and Cl S91 cell lines. CONCLUSION: the new solutions to determinate cell chemosensitivity should be introduced into clinical practice. PMID- 15794337 TI - Crystallization of a non-steroidal anti-inflammatory drug from ethanol-water solution in presence of polymers: physicochemical characterization and release behaviour from suppositories. AB - Altered crystallization condition has been designed and adopted to a model non steroidal anti-inflammatory drug, while crystallizing from ethanol-water solution in absence and presence of polymers such as Eudragit RS and ethylcellulose. To minimize the gastro-intestinal side effects nimesulide was considered as a model drug candidate for the development of suppository formulation. Physicochemical characteristics of the crystals were evaluated by Scanning Electron Microscopy (SEM). X-ray diffraction (XRD) and Fourier Transformed Infrared Spectroscopy (FT IR). Smoothness and sharpness of the crystal have been decreased with increased concentration of a polymer. A little change in crystal habit and geometry has also been observed. Crystals are discrete in nature and more than 90% were in the range of 20-90 micron. The X-ray diffractions of nimesulide crystallized in absence of polymer and physical mixture of drug-polymer revealed fewer high intensity reflections when compared with the drug crystallized in presence of Eudragit RS, which testified a slight decreased ordering of crystal lattice in the latter. In presence of ethylcellulose, slightly increased ordering of crystal lattice was observed. No strong interactions were noticed as revealed by FT-IR spectroscopy. Drug dissolution rate from suppository formulations containing nimesulide crystallized in presence of polymer was found to delay as compared with the suppository prepared by nimesulide crystallized in absence of polymer. PMID- 15794338 TI - Synthesis and antibacterial activity of adamantyl substituted pyrimidines. AB - A series of new S-substituted 6-adamantylpyrimidines were prepared by alkylation on the thiol groups in 6-adamant-1-yl-2-methyl-3H-pyrimidine-4-thione and 6 adamant-1-yl-2-thioxo-2,3-dihydro-1H-pirymidin-4-one, respectively. Then, these compounds were tested against several bacterial strains as well as fungi. PMID- 15794339 TI - Synthesis and anticancer activity of novel 2-amino-4-(4-phenylpiperazino)- 1,3,5 triazine derivatives. AB - The synthesis and anticancer activity of novel 2-amino-4-(4-phenylpiperazino) 1,3,5-triazine derivatives 4a-b and 5a-f are described. Compounds 4a, 5a,c and 5f were evaluated by in vitro assays of growth inhibition against several human tumor cell lines. The vitro cytotoxic activity was found for 2-{2-amino-4-[4-(2 chlorophenyl)piperazino]- 1,3,5-triazin-6-yl}-3-(4-nitrophenyl)acrylonitrile (5f) (IC50 = 0.45 microM - 1.66 microM), whilst other tested compounds were inactive. Some of the molecular orbital calculations for the tested derivatives were also presented. PMID- 15794340 TI - Synthesis and anticonvulsant properties of a series of N-substituted 2-aza spiro[4.5]decane-1,3-diones and 8-phenyl-2-aza-spiro[4.5]decane-1,3-diones. AB - A series of N-phenyl-2-aza-spiro[4.5]decane-1,3-diones [III-VIIII] structurally related to the previously described N-phenyl-3-arylpyrrolidine-2,5-dione (11) was synthesized and tested for their anticonvulsant activity in the maximum electroshock seizure (MES) and metrazole seizure threshold (sc. MET) tests. The most potent of the series were N-(2-methylphenyl)-2-aza-spiro[4.5]decane-1,3 dione [III] and N-(3-methylphenyl)-2-aza-spiro [4.5]decane-1,3-dione [IV], which inhibited seizures in the MES and sc.MET tests. On the other hand, as a preliminary assay we synthesized and tested for the anticonvulsant activity a new N-substituted 8-phenyl-2-aza-spiro[4.5]decane-1,3-dione, containing either a benzyl or a cyclohexyl moiety [IX-XII] at the nitrogen atom. The obtained results showed that the presence and position of the methyl group in the aryl ring [III, IV], as well as an cyclohexane moiety [XI, XIII connected with the imide nitrogen atom, played the essential role for anticonvulsant activity. PMID- 15794342 TI - The influence of prolactin on the chosen biochemical parameters of the rabbit liver in ischemia. AB - An experimental model, based on Pringle's scheme of acute warm hepatic ischemia in normothermia was employed in order to study the hepatoprotective properties of prolactin (PRL). In the proposed model one liver lobe was maintained in the portal circulation and the remaining lobes were perfused with HTK solution for 2 hours. The experiment was carried out on female rabbits of the Chinchilla race. In the control group (n= 10) the liver was perfused with HTK solution. In the examined group (n=10), 3 microg of PRL per g of liver per hour was added to HTK solution. Additionally, the animals in the PRL-treated group were intravenously administered a dose of 600 microg of PRL / kg body weight. 1 h before the surgical treatment. The activities of alanine aminotransferase (ALT), alkaline phosphatase (ALP). gamma-glutamyl transferase (GGT) and the lactate concentration were determined in the eluate obtained from the perfused part of the liver. It was found that administration of prolactin during 2 h of perfusion led to a significant decrease of ALT, ALP and lactate concentrations in the eluate. In addition, increase of calcium concentration in the liver was significantly lower with the prolactin group. The observed results let us to draw the conclusion that administration of PRL shows signs of protective effects on hepatocytes in normothermic acute ischemia. PMID- 15794341 TI - Synthesis and tuberculostatic activity of some (4-phenylpiperazin-1-ylmethyl) 1,3,4-oxadiazole and (4-phenylpiperazin-1-ylmethyl)-1,2,4-triazole derivatives. AB - The phenylpiperazineacetic hydrazide cyclization product. 5-(4-phenylpiperazin-1 ylmethyl)-1,3,4-oxadiazole-2-thiol (1) upon alkylation gave the S-methyl derivative (2). next it was tranmsformed into 1-phenyl-4-(5-pyrrolidin-1-yl-1,3,4 oxadiazol-2-ylmethyl)-piperazine (3). Upon the reaction of I with 1.2 dibromoethane. the derivative (4) containing two joint 1,3,4-oxadiazole systems was obtained. The reactions of compound 1 with chloroacetonitrile and chloroacetic acid gave the corresponding nitrile and carboxylic acid (5, 6) as well. The 4-amino-1,2,4-triazole-3-thiol derivative (7) was obtained in the reaction of compound I with hydrazine hydrate. The phenylpiperazinacetic acid hydrazide was transformed into 1,3,4-oxadiazol-2-ylamine (8), and this, subsequently, into the 2-ethoxy-1,2,4-triazole derivative (9). The compounds obtained were tested in vitro for their tuberculostatic activity. The minimum inhibiting concentrations (MIC) were within 25 - 100 mg/ml. PMID- 15794343 TI - Changes in electric charge and phospholipids composition in erythrocyte membrane of ethanol--poisoned rats after administration of teas. AB - Erythrocyte membrane components and electric charge can be modified by ethanol and by compounds formed as a result of its metabolism, particularly be reactive oxygen species (ROS). The effects of ROS can be neutralized by administering preparations with antioxidant properties. The natural preparations of this kind are teas. For this reason, it has been the purpose of this work to determine effect of green and black tea on membrane electric charge and phospholipids composition of an erythrocyte membrane from rats intoxicated with ethanol. Electrophoresis technique and HPLC have been applied to the above-mentioned studies. Ethanol administration caused an increase in erythrocyte surface charge density and phospholipids composition in the membrane. Administration of green and black tea with ethanol to the rats partially neutralizes the changes provoked by ethanol, and the action of green tea was stronger than that of black tea. PMID- 15794344 TI - [Emergency interventions--catastrophies--fatal treatment errors: dream/trauma for the physician]. PMID- 15794345 TI - [Acute otitis media in childhood: when is symptomatic therapy enough?]]. PMID- 15794347 TI - [Tragic cases of rabies via transplantation: how great is risk of infection for the organ recipient? (interview by Dr. Thomas Meissner)]. PMID- 15794346 TI - [Alleviating pain and preventing inflammation with selective COX-2 inhibitors. Prescribing with confidence in the future]. PMID- 15794348 TI - [Now how does one proceed with diagnosis? Incidental finding of minor increase of liver values]. PMID- 15794349 TI - [Unexplained swelling in the neck. How thorough must the diagnostic workup be?]. PMID- 15794350 TI - [Mass in the neck--the view of the ENT specialist]. AB - A swelling in the neck is a common finding. Underlying causes may be enlarged lymph nodes and cysts, but also benign and malignant tumors. The clinical presentation, however, often gives rise to a fear of a malignant process. In children, more than 95% of such swellings are due to infection. In adults older than 40, however, a malignant tumor may be expected in more that 50% of the cases. Sine early detection is associated with a good prognosis, every mass in the neck must be considered to be potentially malignant. In particular when there is additional difficulty in swallowing or hoarseness, indicative of concomitant involvement of the airways or upper alimentary canal, a further careful diagnostic workup is indicated. Cervical lymph nodes measuring appreciably more than 1 cm in diameter, or additional fever, night sweats or loss of weight must immediately prompt the diagnostic steps necessary to diagnose a malignant lymphoma or carcinoma. PMID- 15794351 TI - [Malignant Hodgkin's and non-Hodgkin's lymphomas]. AB - Over the last twenty years, the number of lymphoma patients in the industrialized countries has increased dramatically. Accordingly the family physician has first contact with patients presenting with swollen cervical glands. To establish a definitive diagnosis, a lymph node must promptly be removed, as completely as possible, for histopathological work-up. Establishment of the diagnosis must be followed by staging and the planning of appropriate (stage-oriented) therapy. Treatment of Hodgkin's lymphoma has an excellent outcome. In early-stage disease, treatment comprises brief polychemotherapy followed by involved-field irradiation; in more advanced stages intensive polychemotherapy is applied. Less satisfactory results are obtained with non-Hodgkin's lymphomas. PMID- 15794352 TI - [Influenza pandemic: A real threat ?]. AB - An influenza pandemic is not merely a theoretical threat, but an historical fact. Reliable predictions of the timing of a new pandemic or its extent are, however, impossible. A pandemic poses a threat to society at large, which can be effectively prepared for by appropriate planning and investment in supplies of vaccine doses, antiviral medication and effective monitoring systems. During the interpandemic phase, general practitioners have an important role in raising the level of immunization and providing medical care to those contracting the disease when a pandemic does break out. PMID- 15794353 TI - [Rapidity of pain relief, medication requirement and patient satisfaction with reflux treatment in the physician's office]. PMID- 15794354 TI - [Electronic health pass, integrated network... more EDV (electronic data processing) doesn't make general practice easier]. PMID- 15794355 TI - [More money than the federal chancellor. Are 260,000 euros too much for a health insurance boss?]. PMID- 15794356 TI - [Patient with beer belly or hypertension. Check for a metabolic syndrome!]. PMID- 15794357 TI - [What the general practitioner can do to rehabilitate the patient]. PMID- 15794358 TI - [Witch milk complication]. PMID- 15794359 TI - [Splenomegaly. Main symptom: often asymptomatic splenic hyperplasia]. PMID- 15794360 TI - [Changes in body composition during an extreme endurance run]. AB - We measured before, during and after a six-day-run in one athlete body weight, skinfold thickness, circumference of extremities, energy expenditure and nutritional intake. Despite an increased fat and protein intake during the race we found a significantly decrease in adipose subcutaneous tissue of the whole body and muscle mass in the active limbs whereas body weight remained stable. We presume that during running as eccentric exercise subcutaneous adipose tissue and muscle mass will be oxidised. PMID- 15794361 TI - [The new features of the bibliography database manager EndNote 6.0 and 7.0]. AB - The bibliography database manager EndNote shows since 1999 a dramatically technical development. A new improved version is published every year. Because the version 7.0 was recently (June 2003) released, we want to describe some aspects of the new version. We also would like to examine, for which user the use of the respective version is recommendable. The use of the software package EndNote 6.0 and also 7.0 for Windows is described. The main reason for getting Endnote 6 is its clearly improved functions and features: Organize a variety of charts equations or pictures and the use of Microsoft-templates. The version 7.0 can be recommended especially to scientist who much works with a Palm particularly. It is also possible to work not only with Microsoft-Word but also with other word processors and creating a bibliography with topic headings. Altogether EndNote 6.0 and also 7.0 provides an excellent combination of features and ease of use. The versions 6.0 or 7.0 are useful especially for people, who every single day use EndNote. EndNote 7.0 is for user of a Palm a special recommendable version. PMID- 15794362 TI - [Fall from a height of 13.5 meters of a child without major injuries]. AB - High-velocity traumas, especially falls from a height, are a major cause of morbidity and mortality in children. Despite the height of fall is a poor predictor of major injury, we must be aware of relevant injuries especially of the head and the musculoskeletal system. The aim is to recognize all relevant problems, or better to avoid the fall. We report a case from a fall of 13.5 meters with an ellbow-fracture and contusions of the head. PMID- 15794363 TI - [Asymptomatic splenomegaly]. PMID- 15794364 TI - [Breast cancer: increased incidence, decreased mortality]. PMID- 15794365 TI - [Aldosterone and aldosterone antagonists. Eplerenone in cardiac insufficiency after myocardial infarction]. PMID- 15794366 TI - [Lyme borreliosis]. PMID- 15794367 TI - [Photosensitization due to drugs: adverse reaction and beneficial effect]. AB - In all areas of everyday life men get in touch with photosensitizers which increase the susceptibility to solar light. Dermal reactions arise after exposure to a photosensitizer and light. Quality and intensity depend on the compound and the radiation dose. Many drugs provoke these responses appearing as an adverse reaction during conventional treatment or as beneficial effect in the context of photodynamic therapy (PDT) or photochemotherapy (PUVA, ECP). Basic mechanisms and clinical aspects are reviewed. PMID- 15794368 TI - [L-arginine. A key substance for dietetic therapy in arteriosclerosis?]. PMID- 15794369 TI - Professional nursing: time for a check-up. PMID- 15794370 TI - Background checks on nurses. PMID- 15794371 TI - Down to the wire: the April 2005 HIPAA deadline. PMID- 15794373 TI - Building a leadership team that works. AB - Radiology administrators often are challenged to do more with less. In today's fast-paced work environment, leaders must be creative. They must surround themselves with good people in order to successfully achieve their organizations' goals. Once a radiology administrator is satisfied and comfortable that he or she has, the right staff involved, a leadership team can be formally establislished. Howard Regional Health System established an Imaging Services Leadership Team with a vision to provide leaders for the staff to "follow," just as team members learn from the radiology administrator. In addition, team members are vital in assisting the radiology administrator in managing the department The process of building the team consisted of 3 steps: selecting team members (the most challenging and time-consuming component), formalizing a functional team, and putting the team into action. Finding the right people, holding regular meetings, and making those team meetings meaningful are keys to a successful leadership team. The implementation of the team has had a positive effect on imaging services: the number of procedures has increased, the team is used as a communication tool for front-line staff, front-line staff are becoming more comfortable with making decisions. PMID- 15794372 TI - Staffing shortages and the theory of constraints. PMID- 15794374 TI - Preparing a cultural strategy for PACS. AB - Health Central Hospital in Ocoee, FL, performs in excess of 100,000 studies per year. In September 2003, it was determined that a picture archiving and communication system (PACS)--already an agenda item set for the current fiscal year-would have a go-live date of no later than July 1, 2004. That meant 9 months to research, develop, and implement the project. The objective was to bring PACS to fruition quickly and smoothly, have buy-in from all parties concerned, and not interrupt the daily flow of patient care in the process. The first step in the project was to select the vendors to be considered. Working closely with the radiology leadership team, a list of physicians who most frequently requested film was compiled. Among them, the team targeted 3 or 4 that it was believed would help champion the effort with their fellow physicians. Along with these pieces, the radiologists, physicians on staff at the hospital, and, of course, the nursing and administrative teams would be an integral part of the project. The groundwork was laid for hiring a consultant to assist in the process. A PACS coordinator job description was created. Then, a staff meeting was called to discuss routine business issues and bring PACS to the table. It is extremely important to bring all relative new data to the staff throughout the project. After that initial meeting, PACS was a regular agenda item. A project team was comprised of 6 persons from radiology, information systems (IS), and administration, as well as a radiologist. Each member was required to make all 4 site visits, give a brief written evaluation of the product seen, and provide a brief narrative for and against the equipment. The vendor was selected, a final proposal was obtained, and contract details were negotiated. A PACS Steering Committee was appointed, and a PACS coordinator and consultant were hired. The PACS was installed, and a computed radiography (CR) system was purchased and integrated with the PACS. Training was paramount to the project's success. The PACS coordinator had experience educating physicians on using the technology; therefore, Health Central was able to provide the staff, radiologists, and hospital physicians with on-site, hands-on training. The hospital's culture has changed since PACS. Complaints have been reduced, and the number of exams waiting for radiologists to read each morning has been reduced by more than 50%. PMID- 15794375 TI - What is IS? PMID- 15794376 TI - Are you coding with grace? PMID- 15794377 TI - Improving medical imaging report turnaround times. AB - Southern Ohio Medical Center (SOMC), a 232-bed community-based teaching hospital, is equipped with state-of-the-art equipment such as 2 16-slice computed tomography (CT) scanners, 3 MR scanners, 3 ultrasound scanners, 2 digital mammography units, and 3 nuclear medicine cameras. One hundred twenty-six employees--ranging from support personnel to technologists along with 7 board certified radiologists--staff the medical imaging department. Procedure volume is approximately 164,000 per year and is performed in all American College of Radiology (ACR)-accredited modalities. Filmless since 1998, SOMC's medical imaging department has resulted in productivity gains to the estimated 164,000 procudures for fiscal year 2005. The catalyst for the department is a robust picture archiving and communication system (PACS). Working with the radiologists, staff, and transcription services, turnaround time was reduced to from 13 hours to 9 hours from exam start to report sign off. Additional technology intervention was essential to further decrease report turnaround time. SOMC served as a beta site for a radiology information system (RIS). The new RIS has allowed the medical imaging department to move from a paper department to a "pseudo paperless" department. Orders, history sheets, consents, and other forms are scanned into the RIS for staff and radiologist use. Requisitions are no longer printed, and staff has access to review workstations to ensure that patients are called back into the department for procedures. This new workflow has also reduced paper traffic within the department. The last piece of the technology puzzle to improve report turnaround time was voice recognition technology. From its implementation, voice recognition enhanced the RIS technology. All of the radiologists began to use the product as soon as it was available. They perform all the editing and corrections either by voice command or by manual typing. The medical imaging department has noted that voice command corrections and editing are more efficient for the radiologist. The overall impact on decreased radiology report turnaround times is not only seen in medical imaging, but also has a global affect within the hospital. SOMC plans to realize a reduction length of patient stays, and a faster process for plotting the course of patient treatment, e.g., faster visits from emergency department (ED) physicians to patients. PMID- 15794378 TI - Perspectives on speech recognition technology. AB - Speech recognition technology is used in all sorts of applications. However, for radiology, the issues are more complex than merely being able to dial a contact from an address book. In addition, radiologists have been hesitant to embrace the technology, with some preferring the status quo. Speech recognition technology has dramatically improved over the past several years, and they generally have been broadly commercialized. However, the use of speech recognition for composition of text reports or email has had only limited acceptance. The overriding reasons appear to be that most computer users prefer not to talk to their computers. They have learned to compose text documents via a "type-and organize" methodology rather than composing the document "in their heads" and dictating. Radiologists are still required to dictate their reports, whether it is digitally, into an analog tape recording device or via a speech recognition system. The benefits extend to the radiologist's patients, and to the radiologist's employers--the hospitals or imaging centers--but it could be said that there is little direct benefit for the radiologist There is a belief that systems should focus more on improving radiologist efficiency rather than emphasizing cost savings and turnaround time. Integration with existing systems is critical. But any technology, in order for it to be well accepted by the primary user, needs to benefit that user. Before selecting any speech recognition technology a radiology administrator should do some research and find answers to several questions that address the basics of speech recognition technology and the companies that provide this technology. In addition, the radiology administrator must ensure that the facility is prepared to implement the technology and address any workflow- or culture-related issues that may arise. There are a number of opportunities for improvement in speech recognition radiology applications. These include the ongoing need for improvement recognition rates, the need to streamline integration with picture archiving and communication system (PACS) and radiology information system (RIS) technologies, and the general need to improve the user interface. In addition to these improvements, one can expect an increased adoption of structured reporting technologies within radiology. These techniques allow easier automated extraction of content and more flexible communication and organization of data (such as communication to electronic medical record systems). PMID- 15794379 TI - Turf wars. PMID- 15794380 TI - New staff utlization survey provides data on staffing levels, FTE volume. PMID- 15794381 TI - ER: the front door to the hospital. PMID- 15794382 TI - Look, mom! I made an imaging center! PMID- 15794383 TI - Microglial imaging with positron emission tomography and atrophy measurements with magnetic resonance imaging in multiple sclerosis: a correlative study. AB - OBJECTIVE: The objectives of the present study were to assess brain atrophy in multiple sclerosis (MS) patients during different disease stages and to investigate by PET and [11C]PK11195, a marker of microglial activation, the relationship between inflammation, atrophy and clinically relevant measures. METHODS: Eight healthy subjects and 22 MS patients were included. Semiquantitative [11C]PK11195 uptake values, with normalization on cortical grey matter, were measured for magnetic resonance imaging T2- and T1-lesions and normal appearing white matter (NAWM). As atrophy index we used the ratio of the amount of white and grey matter divided by the ventricular size, using an optimized a priori based segmentation algorithm (SPM99). RESULTS: Atrophy was significantly greater in MS patients compared to age-matched controls. A significant correlation was found between brain atrophy and both disease duration and disability, as measured with the Expanded Disability Status Scale. For NAWM, [11C]PK11195 uptake increased with the amount of atrophy, while T2-lesional [11C]PK11195 uptake values decreased according to increasing brain atrophy. CONCLUSIONS: The present study suggests that brain atrophy, correlating with disease duration and disability, is directly related to NAWM and T2-lesional inflammation as measured by microglial activation. PMID- 15794384 TI - Tumefactive demyelinating lesions: conventional and advanced magnetic resonance imaging. AB - In rare instances, demyelinating disorders present with radiological features that mimic a brain tumour. This often leads to biopsy, which--apart from carrying significant morbidity--frequently turns out as nondiagnostic or dispensable. We therefore set out to assess the contribution of repeated conventional magnetic resonance imaging (MRI), 1H-MR spectroscopy and magnetization transfer imaging in establishing a correct diagnosis of tumefactive demyelinating lesions (TDLs). We studied two females and one male, who presented with TDLs that led to brain biopsy in two cases, for up to three years. TDLs were characterized by the following features: (a) delayed or absent response to high-dose steroids together with progressive lesion growth over several weeks; (b) late or sparse enhancement, ill-defined borders, signal inhomogeneity and considerable concomitant oedema; and (c) normalization of initial increases in lipid and lactate peaks within three to four weeks, followed by persistent, marked reductions of the neuronal marker NAA and MTR values around or below 30%. These imaging characteristics reflected the histological correlate of marked demyelination in the absence of significant inflammation. MRI techniques thus appear to have the potential to establish a correct diagnosis of this subtype of TDLs. Awareness of these possibilities might obviate the need for biopsy at least in some cases in future. PMID- 15794385 TI - Brain atrophy and magnetization transfer ratio following methylprednisolone in multiple sclerosis: short-term changes and long-term implications. AB - BACKGROUND: The short-term effect of corticosteroids on MRI measures of multiple sclerosis (MS) is not well understood and may have a significant impact when using these quantitative measures to evaluate disease activity and changes following other therapeutic interventions. OBJECTIVE: To determine the impact of a course of intravenous methylprednisolone (IVMP) on quantitative measures of disease activity and tissue injury in MS patients. METHODS: We prospectively measured brain parenchymal fraction (BPF), magnetization transfer ratio (MTR, lesional and whole brain), and lesion volumes on nine weekly brain MRI studies in ten MS patients receiving a course of IVMP. A group of nine MS patients not receiving IVMP served as controls. RESULTS: In comparison to untreated controls, BPF declined over the eight weeks following IVMP treatment (P <0.02). BPF decline was most prominent in patients with secondary progressive MS (SPMS, P <0.03), and was not seen in relapsing-remitting (RR) MS patients. Short-term change in BPF correlated with baseline BPF (r =0.62, P =0.05) and short-term change in lesional MTR (r = -0.55, P =0.03), but not with change in enhancing lesion volume. Short term change in lesional MTR inversely correlated with baseline lesional and whole brain MTR (r = -0.79, P =0.04 for both). There was no significant difference between treated and control patients in measures of MTR or T2, T1 or enhancing lesion volumes. CONCLUSIONS: Patients with SPMS showed a greater decline in BPF following IVMP than RRMS patients. A correlation between changes in BPF and MTR suggest that these changes are secondary to altered water content within MS lesions. Differential response to a standardized therapeutic intervention in RRMS and SPMS suggests that responses to therapy may differ due to a fundamental pathologic difference between early and late stage MS. PMID- 15794386 TI - Contrast-enhanced MRI lesions during treatment with interferonbeta-1b predict increase in T1 black hole volume in patients with relapsing-remitting multiple sclerosis. AB - T1 black holes (BH) have been found to represent focal areas of substantial central nervous system tissue damage in multiple sclerosis (MS) patients. We examined the development of T1 BH over a three-year period of treatment with interferon (IFN)beta-1b in a group of 20 patients with relapsing-remitting MS. The number of contrast-enhancing lesions (CEL) after one year of treatment predicted a change in the T1 BH volume in the following two years. In patients without CEL, the T1 BH volume remained stable, whereas it increased in patients with CEL. The occurrence of CEL in patients treated with IFNbeta may indicate a heightened risk of accumulating T1 BH. PMID- 15794387 TI - A double-blind, randomized, controlled study of oral pirfenidone for treatment of secondary progressive multiple sclerosis. AB - Currently, there are no approved treatments for secondary progressive multiple sclerosis (MS) that stabilize or reverse the neurological disabilities associated with this disease. Oral pirfenidone was found to stabilize and overcome the disabilities in two published independent open-label studies in secondary progressive MS. This led us to study pirfenidone in a phase II double-blind, randomized and controlled, clinical trial in patients with advanced secondary progressive MS for 12 months. Forty-three patients met the eligibility criteria approved by the IRB and accepted by the FDA. Of these patients, 18 were randomly assigned to placebo and 25 patients to oral pirfenidone groups. All eligible patients were included in the statistical analysis of the data according to intention-to-treat principles. Some patients on oral pirfenidone manifested mild drug-related adverse effects, but it was well tolerated overall. By one month, pirfenidone significantly (P < 0.05) improved the Scripps Neurological Rating Scale (SNRS) scores, and scores remained significantly improved for 3, 6 and 12 months when compared to the baseline SNRS scores. In contrast, the SNRS scores of patients on oral placebo were not significantly improved at 1, 3, 6 or 12 months of the study, when compared with baseline scores. Oral pirfenidone significantly (P <0.04) reduced the incidence of relapses (27.8% on placebo versus 8.0% on pirfenidone). Furthermore, oral pirfenidone treatment was associated with a marked improvement in bladder dysfunction (40.0% on pirfenidone versus 16.7% on placebo). Expanded Disability Status Scale scores and MRI lesion count were not significantly different in the placebo and pirfenidone groups. These findings indicate a significant effect of pirfenidone on clinical disability and bladder function for secondary progressive MS patients. A major multicentre, double blind, randomized, controlled trial is justified. PMID- 15794388 TI - Lipoic acid in multiple sclerosis: a pilot study. AB - Lipoic acid (LA) is an antioxidant that suppresses and treats an animal model of multiple sclerosis (MS), experimental autoimmune encephalomyelitis. The purpose of this study was to determine the pharmacokinetics (PK), tolerability and effects on matrix metalloproteinase-9 (MMP-9) and soluble intercellular adhesion molecule-1 (sICAMP-1) of oral LA in patients with MS. Thirty-seven MS subjects were randomly assigned to one of four groups: placebo, LA 600 mg twice a day, LA 1200 mg once a day and LA 1200 mg twice a day. Subjects took study capsules for 14 days. We found that subjects taking 1200 mg LA had substantially higher peak serum LA levels than those taking 600 mg and that peak levels varied considerably among subjects. We also found a significant negative correlation between peak serum LA levels and mean changes in serum MMP-9 levels (T = -0.263, P =0.04). There was a significant dose response relationship between LA and mean change in serum sICAM-1 levels (P =0.03). We conclude that oral LA is generally well tolerated and appears capable of reducing serum MMP-9 and sICAM-1 levels. LA may prove useful in treating MS by inhibiting MMP-9 activity and interfering with T cell migration into the CNS. PMID- 15794389 TI - A randomized, double-blind, placebo-controlled study of oral hydrolytic enzymes in relapsing multiple sclerosis. AB - Oral administration of hydrolytic enzymes (HE), such as bromelain, trypsin and rutosid, may have beneficial effects on the clinical course of neurological symptoms related to multiple sclerosis (MS). This is supported by a complete protection by HE from experimental allergic encephalomyelitis, an animal model related to MS. Three hundred and one patients with relapsing MS were enrolled in a double-blind, placebo-controlled trial. No treatment effect between the placebo and the HE groups was found either for clinical or MRI parameters. PMID- 15794390 TI - Optimization of the safety and efficacy of interferon beta 1b and azathioprine combination therapy in multiple sclerosis. AB - We conducted an open-label pilot clinical trial to evaluate the safety and efficacy of adding oral azathioprine to the treatment regimen of 15 multiple sclerosis patients breaking through monotherapy with interferon beta-1b. There were no serious adverse events. Gastrointestinal side effects and leukopenia were the most common adverse events and limited dose escalation. There was a 65% reduction in the number of gadolinium-enhanced magnetic resonance imaging (MRI) lesions on combination therapy compared to the baseline values (P =0.003). A total WBC count less than 4800/mm3 was the best predictor of MRI response. PMID- 15794391 TI - The relationship between depression and interferon beta-1a therapy in patients with multiple sclerosis. AB - It has been suggested that interferons (IFN) may cause depression de novo or worsen pre-existing depression. Depression data collected using validated instruments from individual clinical trials in multiple sclerosis, however, have consistently failed to identify an association. In this study, pooled data from 6 controlled studies and 17 noncontrolled clinical trials of subcutaneous IFN beta 1a were assessed to determine the relationship between IFN therapy with physician reports of depression and suicide. In distinction to the negative findings for depressive symptom ratings, pooling of physician-reported side effect data from these clinical trials identified a statistically significant association between depression and IFN use during the first six months of treatment There was an association between these reported episodes of depression and discontinuation of IFN therapy, but IFN treatment was not associated with suicide attempts. IFN beta 1a may induce a constellation of symptoms, particularly early in therapy, that may be labelled as depression by physicians. However, the lack of an increase in depression-rating scale scores and the lack of association with suicide risk suggests that the syndrome may be an atypical one. PMID- 15794393 TI - Depression and fatigue in relapsing-remitting MS: the role of symptomatic variability. AB - Depression and fatigue are common psychosocial sequelae of MS. An infrequently examined issue in the MS literature is the effect of MS-related symptom variability on incidence of depression and fatigue. The current study was designed to examine the relationship of variability in physical, cognitive, and social/environmental functioning with depression and fatigue in MS. Forty-eight relapsing-remitting MS patients from a larger sample completed self-report measures of depression and fatigue. They were also administered a structured interview that assessed current, best, and worst levels of functioning since being diagnosed with MS. Higher levels of symptom variability since disease onset were associated with depression and fatigue, even after accounting for MS-related physical disability. Regression analyses indicated that variability in social and environmental functioning was particularly associated with depression, and variability in physical abilities was associated with fatigue. These findings suggest that a more variable course of MS symptoms is associated with increased depression and fatigue. Focused assessment of variability in symptoms may be useful in detecting and subsequently treating depression and fatigue in MS patients. PMID- 15794392 TI - Self-injection anxiety training: a treatment for patients unable to self-inject injectable medications. AB - Anxiety and phobia frequently prevent patients with multiple sclerosis (MS) from self-injecting their injectable disease-modifying medications. This small, randomized, controlled trial tested the efficacy of a six-session nurse administered programme to teach self-injection to patients with MS, who, due to anxiety or phobia, were unable to self-inject their injectable medications. Participants were 30 patients with MS who were prescribed interferon beta-1a (IFNbeta-1a) administered via weekly intramuscular injection. All patients were unable to self-inject due to anxiety or phobia. Patients were randomized to either the six-session Self-Injection Anxiety Therapy (SIAT) or a control telephone support condition modelled on the support programme offered by the manufacturer of IFNbeta-1a. Four patients dropped out of SIAT while three dropped out of the control condition. Eight patients receiving SLAT compared to three control patients, were able to self-inject after six weeks of treatment. SIAT patients were significantly more likely to achieve self-injection at treatment cessation, compared to telephone control patients, in completer analyses (p =0.022), however, this only reached a trend in intent-to-treat analyses (p =0.058). These findings suggest that SIAT is a potentially valuable intervention to teach self-injection skills to injection phobic and anxious patients, and should be investigated more thoroughly in a larger clinical trial. PMID- 15794394 TI - Pattern of neuropsychological impairment in the early phase of relapsing remitting multiple sclerosis. AB - To investigate the neuropsychological profile in the first few years post-onset of relapsing-remitting multiple sclerosis (MS) we carried out a comprehensive neuropsychological evaluation of 33 patients characterized by very short evolution of this disease, minimal levels of neurological disability and preserved general cognition. Thirty-three individually pair-matched controls were also evaluated. Patients performed as well as controls on many of the cognitive exploration measures. Nevertheless, the group of patients evinced a general slowness that affected motor execution and cognitive processing. Memory functions were characterized by preservation of working memory, retrieval or storage of information and a deficit at the acquisition phase in (verbal and visual) supraspan tasks. In addition, significant correlations were observed between some measures of information processing speed and memory. These results highlight the importance of studying cognitive deficits not only in the different subtypes of MS but also in different phases of the disease. PMID- 15794395 TI - Does the Modified Fatigue Impact Scale offer a more comprehensive assessment of fatigue in MS? AB - BACKGROUND: As a symptom of multiple sclerosis (MS), fatigue is difficult to manage because of its unknown etiology, the lack of efficacy of the drugs tested to date and the absence of consensus about which would be the ideal measure to assess fatigue. OBJECTIVE: Our aim was to assess the frequency of fatigue in a sample of MS patients and healthy controls (HC) using two fatigue scales, the Fatigue Severity Scale (FSS) and the Modified Fatigue Impact Scale (MFIS) with physical, cognitive and psychosocial subscales. We also studied the relationship fatigue has with depression, disability and interferon beta. METHODS: Three hundred and fifty-four individuals (231 MS patients and 123 HC) were included in this cross-sectional study. Fatigue was assessed using the FSS and MFIS. Depression was measured by the Beck Depression Inventory (BDI), and disability by the Expanded Disability Status Scale (EDSS). A status of fatigue was considered when the FSS > or =5, of non-fatigue when the FSS < or =4, and scores between 4.1 and 4.9 were considered doubtful fatigue cases. RESULTS: Fifty-five percent of MS patients and 13% of HC were fatigued. The global MFIS score positively correlated with the FSS in MS and HC (r =0.68 for MS and r =0.59 for HC, p <0.0001). Nonetheless, the MFIS physical subscale showed the strongest correlation score with the FSS (r =0.75, p <0.0001). In addition, a prediction analysis showed the physical MFIS subscale to be the only independent predictor of FSS score (p <0.0001), suggesting other aspects of fatigue, as cognition and psychosocial functions, may be explored by the FSS to a lesser extent. Depression also correlated with fatigue (r =0.48 for the FSS and r =0.7 for the MFIS, p <0.0001) and, although EDSS correlated with fatigue as well, the scores decreased after correcting for depression. Interferon beta showed no relationship with fatigue. CONCLUSIONS: Fatigue is a frequent symptom found in MS patients and clearly related with depression. Each fatigue scale correlates with one another, indicating that they are measuring similar constructs. Nevertheless, spheres of fatigue as cognition and psychosocial functions are probably better measured by the MFIS, although this hypothesis will need to be confirmed with appropriate psychometrical testing. PMID- 15794396 TI - The relationship between self-awareness of neurobehavioral symptoms, cognitive functioning, and emotional symptoms in multiple sclerosis. AB - OBJECTIVE: To examine self-awareness of neurobehavioral symptoms in multiple sclerosis (MS) across three domains of function (apathy, disinhibition, and executive control), and examine the relationship between self-awareness and cognitive functioning. METHODS: Twenty-six individuals with MS completed neuropsychological testing, measures of emotional functioning, and self-ratings of neurobehavioral symptoms using the Frontal Systems Behavior Scale (FrSBe), a 46-item questionnaire with subscales assessing frequency of symptoms in executive dysfunction, disinhibition, and apathy. Informants' ratings of the FrSBe were also obtained. Decreasing differences between patient and informant reports on each subscale of the FrSBe (concordance) indicate higher levels of self awareness. RESULTS: Results showed significant positive correlations between cognitive abilities and self-awareness of executive dysfunction and disinhibition. In contrast, affect symptomatology (measures of anxiety and depression) were negatively correlated with self-awareness of executive dysfunction. CONCLUSIONS: Level of self-awareness of neurobehavioral symptoms in MS is related to level of cognitive impairment In addition, symptoms of depression and anxiety reduced the accuracy of self-reporting. Thus, a clinician who relies on self-reports in creating an evaluation and treatment plan should consider the patient's cognitive and emotional states. PMID- 15794397 TI - Spiroergometric and spirometric parameters in patients with multiple sclerosis: are there any links between these parameters and fatigue, depression, neurological impairment, disability, handicap and quality of life in multiple sclerosis? AB - One-hundred and twelve patients with multiple sclerosis were selected as population-based sample and examined on impairment (Expanded Disability Status Scale), disability (Barthel Index), handicap (Environment Status Scale), the quality of life (Multiple Sclerosis Quality of Life), fatigue (Modified Fatigue Impact Scale), depression (Beck Depression Inventory Score), respiratory function (spirometric parameters on spirometry) and physical fitness (spiroergometric parameters on a bicycle ergometer). The aim of the study was to examine and analyse (descriptive statistics) spiroergometric and spirometric parameters in patients with multiple sclerosis. Firstly, we tested the hypothesis whether spiroergometric and spirometric parameters are decreased and whether there are any correlations between these parameters and measures of impairment, depression, disability, handicap and quality of life. Secondly, we tested the hypothesis whether there is any correlation between a possible deconditioning and fatigue, and between a possible respiratory dysfunction and fatigue in multiple sclerosis. It results from this study that many spiroergometric parameters in patients with multiple sclerosis are significantly lowered in comparison to the population norm. A link can be found between some spiroergometric parameters and neurological impairment, disability, handicap and quality of life. It is not possible to prove any correlation between spiroergometric parameters and depression. From the spirometric parameters, these are expiratory flows that are significantly lowered in MS patients. It is not possible to prove any correlation between spirometric parameters and fatigue, depression, neurological impairment, duration of the disease, disability, handicap and quality of life in multiple sclerosis. PMID- 15794398 TI - Discrepancies in the interpretation of clinical symptoms and signs in the diagnosis of multiple sclerosis. A proposal for standardization. AB - The new McDonald diagnostic criteria for multiple sclerosis (MS) incorporate detailed criteria for the interpretation and classification of magnetic resonance imaging (MRI) findings, but, in contrast, provide no instructions for the interpretation of clinical findings. Because MS according to the McDonald criteria is one of the primary endpoints in a large trial enrolling patients after the first manifestation suggestive for a demyelinating disease (BENEFIT study), it was decided to organize a centralized eligibility assessment for this trial. During this eligibility assessment it was observed that there were marked inconsistencies in the decisions of participating neurologists with respect to the classification of clinical symptoms as being caused by one or more lesions provoking discussions in about one in every five patients. This paper describes these inconsistencies and their sources, and recommends a systematic approach that attempts to reduce the variability in interpreting clinical findings. PMID- 15794399 TI - Cost of multiple sclerosis by level of disability: a review of literature. AB - We performed a review of the economic literature to identify what is known about the relationship between Expanded Disability Status Scale (EDSS) categories and cost of multiple sclerosis (MS). We sought cohort studies of patients with multiple sclerosis that described the costs attributed to each EDSS score and utilized specific inclusion criteria for the selection of 10 studies. We found that both direct and indirect costs rise continuously with increasing EDSS category, and this rise is qualitatively exponential. The rise in indirect costs appears at lower EDSS scores. The cost of a relapse occurring in any given EDSS category exceeds that associated with that particular EDSS category. Few studies comprehensively assessed the entire spectrum of the costs, and much of the literature is based on EDSS categories in coarse groupings. In spite of several variations between studies, one important conclusion that we can draw is that rise in cost is positively correlated to scores on the EDSS categories, and therefore agents with a capacity to prevent or arrest the rate of MS progression may affect the overall cost of MS. PMID- 15794400 TI - Factors limiting maximal oxygen uptake in exertional monoparesis. AB - Strength and maximal oxygen uptake (VO2max) for each leg were compared in a trained multiple sclerosis (MS) patient with exertional left monoparesis. Left quadriceps strength and left leg VO2max were 22% and 30% lower, respectively, compared to the right (control) leg. Because the same O2 delivery system served each leg during exercise, VO2max of the paretic leg was not limited by cardiorespiratory factors but rather by strength and/or muscle oxidative capacity. However, training with monoparesis likely enhanced O2 extraction and aerobic work capacity of the right leg. PMID- 15794401 TI - Cerebral venous thrombosis and demyelinating diseases: report of a case in a clinically isolated syndrome suggestive of multiple sclerosis onset and review of the literature. AB - Cerebral venous thrombosis (CVT) has been described in several cases of clinically definite multiple sclerosis (MS). In the majority of these, lumbar puncture followed by intravenous corticosteroid treatment was suspected as the cause. We report what is, to our knowledge, the first case of a patient with a multifocal clinically isolated syndrome suggestive of MS onset, who developed multiple CVT after lumbar puncture and during high-dose i.v. corticosteroid treatment We conclude that the sequence 'lumbar puncture followed by corticosteroid treatment' may be a contributory risk factor for the development of CVT when associated with other risk factors. PMID- 15794402 TI - Jessner-Kanof lymphocytic infiltration of the skin associated with glatiramer acetate. AB - Glatiramer acetate (GLAT) is a well tolerated and safe immunomodulatory drug for the treatment of relapsing-remitting multiple sclerosis. The most commonly recognized side effects are localized injection site reactions consisting of pain, pruritus, mild erythema and induration, which sometimes persist for several days. We describe the first case of a biopsy-proven lymphocytic infiltration (T cell pseudolymphoma) with the clinical appearance of a figured erythema on the ventrolateral thighs in the first four weeks under GLAT treatment, resolving without any evidence of recurrence despite ongoing therapy. A T-cell pseudolymphoma is a very rare side effect of GLAT treatment. For clinical purposes it is important to state that re-exposition after GLAT-induced pseudolymphoma is possible without permanent sequelae. PMID- 15794403 TI - MSRV, Syncytin and the role of endogenous retroviral proteins in demyelination. PMID- 15794404 TI - [Total control of asthma]. PMID- 15794407 TI - [Sublingual immunotherapy with allergenic extract of Dermatophagoides pteronyssinus in asthmatic children]. AB - BACKGROUND: Sublingual immunotherapy is based on the treatment of distinct pathologies. It is not exempt from discomfort and hazards; thus, alternative approaches for therapy are being tested. OBJECTIVE: To assess the effectiveness of sublingual immunotherapy, with Dermatophagoides pteronyssinus in asthmatic children. MATERIAL AND METHODS: Fifty pediatric patients of both sexes between 5 and 15 years were randomly divided into two groups. The first group received sublingual immunotherapy with Dermatophagoides pteronyssinus in concentrations of 500, 1000, 2000, 5000, 8000 and 10,000 biological units. Both groups received conventional therapy, and spirometry and total IgE were measured every 30 days. The use of bronchodilators and steroids was assessed before and during the test. RESULTS: Peak expiratory flow improved in the study group with a remarkable difference (p < 0.05). There were not significant differences in the control group (p > 0.05); however, at the end of the study an increase of 80% was observed. In urgency services, patients who received immunotherapy had relative risk (RR) = 0.36 and confidence interval (CI) of 95%, 0.15, 0.84. The use of steroids equally decreased in patients receiving immunotherapy RR = 0.27, CI 95%, 0.10, 0.69. CONCLUSIONS: Sublingual immunotherapy does reduce the risk for asthma episodes in children. PMID- 15794406 TI - [Lung functioning changes of asthmatic adults in treatment with cromolyn sodium and beclomethasone]. AB - BACKGROUND: In asthmatic patients, besides symptoms, international guidelines recommend the measure of lung functioning tests as a parameter leading to diagnostic classification and evaluation of the treatment's effect. OBJECTIVE: To determine the changes in the lung function of asthmatic adults in treatment with cromolyn sodium and beclomethasone. MATERIAL AND METHODS: We made a clinical trial in adults with more than five years of persistent moderate asthma diagnosis. Before beginning the treatment there were carried out pulmonary function tests (PFR) with basal espirometry (prebeta2) and postbronchodilatador (posbeta2). To receive treatment, the patients were randomized in group A (beclomethasone), B (cromolyn sodium) and C (both medications). Treatment was completed by 6 months and at the end the spirometry was repeated. The results were analyzed with ANOVA, Student t and Mann-Whitney U tests according to the variables distribution. RESULTS: In each group 15 patients were included. The age, the asthma evolution time and the gender relationship were without differences. The results of the PFR are described as the average of the differences among the initial values and those obtained six months later: FEF25 75: prebeta2: group A: 15.4, B: 3.6 and C: 13.8 (ANOVA, p < 0.01); posbeta2: group A: 11, B: 4.5 and C: 17.5 (ANOVA, p < 0.5). FEF50: pre32: group A: 8.4, B: 2.3 and C: 14.7 (ANOVA, p <0.5); posbeta2: group A: 0.2, B: 3.1 and C: 15.7 (ANOVA, p < 0.01). FEF75: prebeta2: group A: 8.4, B: 4.7 and C: 13.6 (ANOVA, p < 0.5); posbeta2: group A: 4.2, B: 6.7 and C: 19.4 (ANOVA, p < 0.01). The differences between groups were: FEF25-75: prebeta2: A vs B, p = 0.08; B vs C, p = 0.1 and A vs C, p = 0.5; posbeta2: A vs B, p = 0.06; B vs C, p = 0.005 and A vs C, p =0.2. VF50: prebeta2: A vs B, p = 0.1; B vs C, p = 0.02 and A vs C, p = 0.1; posbeta2: A vs B, p = 0.4; B vs C, p = 0.001 and A vs C, p = 0.008. VF75: prebeta2: A vs B, p = 0.4; B vs C, p =0.03 and A vs C, p = 0.08; posbeta2: A vs B, p = 0.1; B vs C, p = 0.02 and A vs C, p =0.003. CONCLUSIONS: In the asthmatic adult of long evolution, the inhaled treatment with cromolyn sodium improves the pulmonary function. That improvement is better with the use of beclomethasone, but the combined use improves even more the spirometry parameters. PMID- 15794405 TI - [Autoimmune urticaria. Treatment with methotrexate]. AB - BACKGROUND: It has been established that 27-50% of patients with idiopathic chronic urticaria have antibodies directed against the alpha chain of the high affinity IgE receptor, which are indirectly detected by cutaneous tests with autoserum. Thus, an autoimmune urticaria diagnosis can be settled. OBJECTIVE: To prove methotrexate's efficiency in patients with autoimmune urticaria. MATERIAL AND METHODS: Seven patients took part in the study. A series of tests was performed in order to rule out any possible infectious, metabolic, or physical etiology. Initial treatment with methotrexate with doses of 2.5 mg every 12 hours, two days a week was provided. In case there were no toxicity data, doses would increase to three days a week for a 6-week period. RESULTS: Statistically significant improvement was observed in the itching, as well as the presence of spots, repercussion on daily activities, sleep disorders. There was no statistical difference regarding the extension of the lesions and the presence of angioedema. Adverse effects were not significant. CONCLUSION: We conclude that methotrexate is effective in the treatment of autoimmune urticaria. PMID- 15794409 TI - [Status asthmaticus vs transtracheal membrane and cyclic neutropenia: a report of a case]. AB - INTRODUCTION: Acute episodes of asthma are one of the most common respiratory emergencies, and status asthmaticus is the most severe respiratory impairment. Because "not all the wheezes are manifestations of asthma", the clinician needs to consider non asthmatic causes of wheezing in the differential diagnosis such as tracheal stenosis. CLINICAL CASE: 7 years old male, hispanic, non atopic background, with two previous cases of acute asthma. Current condition: 15 days of cough, dyspnea and wheezes, without fever. The physical examination revealed nasal flutter, use of accessory muscles of respiration, fatigue and bibasal hypoventilation. Mitomycin was applied during nasolaryngoscopy. He was discharged 17 days after with tracheal stenosis diagnosis. Biopsy reported respiratory epithelium with chronic inflammation and fibrosis. Six dilatations were performed because of persistent stenosis and he is being evaluated by infectologist and hematologist for repeated upper airway infections associated with cyclic neutropenia. PMID- 15794410 TI - [Is it possible to teach ethical values?]. PMID- 15794408 TI - [Prevalence and potential triggering factors of chronic urticaria and angioedema in an urban area of northeastern Mexico]. AB - BACKGROUND: Urticaria and angioedema are illnesses of easy diagnosis, but to solve them or to determine their cause is complex, as well as the therapeutic management of chronic cases. OBJECTIVE: To determine the prevalence, possible precipitating factors and chronic urticaria characteristics. MATERIAL AND METHODS: A questionnaire was applied to 4,000 persons selected at random, from the ages of 18-50 years, women and men. We made up simple frequencies, odds ratio, confidence interval at 95%. Possible interventions from the some variables were determined through regression logistic model. RESULTS: Chronic urticaria and angioedema prevalence was 4.8%. From the 147 sick persons, 71.4% were women (p = 0.00) 51.70% had only wheals, 10.88% angioedema alone and 37.41% of the patients with chronic urticaria also have angioedema. Potential triggering factors that we found were 18.36% drugs (p = 0.00) and 4.08% foods (p = 0.00). Family (p = 0.00) and personal allergy (p = 0.00) antecedents were documented in 55.8% and 68.70%, respectively. In 40.1% symptoms interfere with daily activities of the patients and 53.1% of patients had symptoms at night. CONCLUSIONS: Despite of being a retrospective study, the results show us that the chronic urticaria and angioedema are important health problems in the northeastern of Mexico. We can not identify the etiology, and the symptoms can disturb quality of life of the patients. Female sex, family and personal allergy antecedent could be important for these pathology presentations. PMID- 15794411 TI - [Scholar absenteeism and use of health services in children and adolescents with wheezing]. AB - INTRODUCTION: Asthma is an important problem at the primary care level where almost 97% of the cases are managed. Brazil shows high rates of the disease nevertheless few is known about the characteristics related to school absenteeism and the health care usage among urban pediatric population of Duque de Caxias County, Rio de Janeiro. OBJECTIVES: To obtain an estimate of the prevalence of school absenteeism and to evaluate the use of health care related to wheezing in asthmatic and non asthmatic children and adolescents. MATERIAL AND METHODS: An observational cross-sectional study was carried out using a questionnaire for a prospective data collection. The gold standard for asthma was a minimal increment of 12% in the forced expiratory volume of the first second (FEV1), after a bronchodilatation test. RESULTS: 211 patients seeking emergency care for a number of conditions with ages between 5 and 15 years were evaluated. Sample asthma prevalence and reported wheezing rates were: 22% (47/211) and 47% (98/211). About 67% (141/211) of the families had an annual income lesser than US dollar 3600. Asthmatic and non-asthmatic patients showed, respectively, the following averages and rates: 1) emergency department visits: 40.4% (19/47) and 29.3% (48/164), 2) ambulatory visits: 34% (16/47) and 14.6% (24/164) (p = 0.003), 3) hospitalizations: 6.4% (3/47) and 4.9% (8/164), 4) number of consultations in ambulatory: 1.74 (SD = 2.71) and 0.47 (SD = 1.44) (p = 0.00003), 5) number of visits in emergency department: 1.47 (SD = 2.36) and 0.95 (SD = 2.15). CONCLUSION: The results are relevant as they describe asthma morbidity, showing that wheezing is a heavy burden to the population of Duque de Caxias. Further studies are needed to better evaluate the group of children and adolescents that although being classified as non-asthmatic presented some characteristics that were to the asthmatic patients. PMID- 15794412 TI - [Therapeutic effects of an antileukotriene in children with moderate-severe persistent asthma]. AB - BACKGROUND: Asthma is a chronic inflammatory disease of the airways, in which participate many cells and cellular elements. OBJECTIVE: To assess the utility of the zafirlukast, an antagonist of the receivers of the leukotrienes, in asthmatic children of our environment and to evaluate the grade of safety of the medication in the preventive treatment of the asthma. MATERIAL AND METHODS: We selected 50 moderate-severe asthmatic children, between 12 and 16 years (of both sexes) with non satisfactory evolution, in spite of the conventional preventive treatment for the asthma. Zafirlukast were given in dose of 20 mg every 12 hours, far from the foods, during 12 weeks. We carried out laboratory studies, before and after the treatment, evaluating the hematic (Hb and Hto), renal (creatinine) and hepatic (TGP) functions. They were continued in allergy consultation at the 4, 8, and 12 weeks of treatment, to know their clinical evolution, the apparition of adverse reactions and the use of other asthmatic medications in the period of the treatment. RESULTS: The 88% of the children improved clinically. The patients decreased the use of oral steroids and only 4 adverse reactions (8%) were reported, with total regression of the symptoms when suppressing the medication. The averages of the laboratory results, before and after the treatment, reflected values into the established normal limits, Hb: 12.4 (before) and 12.1 g/L (later), creatinine: 63.2 and 67.1 mMol/1 and TGP: 2.1 and 3.6 units. CONCLUSIONS: We observed in our study that the preventive treatment with Zafirlukast is useful and well tolerated, representing a new expectation for the asthma, in pediatric ages. PMID- 15794413 TI - Toll-like receptors: dysregulation in vivo in patients with acute respiratory distress syndrome. AB - OBJECTIVE: To determine the time course of the expression of five toll-like receptors (TLRs 1-5) in mixed blood mononuclear cells and their relationship to pro-inflammatory and anti-inflammatory cytokines during acute respiratory distress syndrome (ARDS). MATERIAL AND METHODS: In a prospective, a longitudinal study was done at an intensive-care unit of a university-affiliated hospital. Seven consecutive patients with ARDS were studied. We followed the onset and progression of ARDS, and subsequent patient recovery or death, and compared patient data with data from a group of healthy volunteers. We separated mixed blood mononuclear cells using Ficoll-Hypaque to detect the transcripts of human TLRs 1-5. TLR mRNAs were isolated by semiquantitative reverse-transcription PCR (RT-PCR). Each signal was expressed as the ratio of TLR mRNA to beta-actin. Cytokines, including tumor necrosis factor alpha, interferon gamma, and interleukins 4, 6, 8, 10, and 12, were assayed using commercial ELISA kits. RESULTS: Dysregulation in the transcription of TLRs gene, principally in ARDS surviving patients, was observed. Down-regulated expressions of TLR1, TLR4, and TLR5 mRNAs were observed in the first 24 h in patients who survived, and probably played a key role in the survival of patients with ARDS caused by sepsis. Serum levels of cytokines such as IL-6, 8 and 10 were significantly increased in patients with ARDS as compared with levels in the healthy volunteers. However, serum levels of IL-12 were lower in patients with ARDS than in the healthy volunteers. There was difference in serum cytokines concentration between survivors and non-survivors patients with ARDS (except for interleukin-10). CONCLUSIONS: Thus, TLRs gene dysregulation and cytokine profiles are probably important prognostic factors for patient outcome and survival after ARDS. PMID- 15794414 TI - [Helicobacter pylori and disease]. AB - Helicobacter pylori infection is considered an important health problem due to the fact that there are reports that describe that more than 50% of the world's populations is affected by it. The infection has been associated with multiple diseases, gastric and extra-gastric, whose signs and symptoms are overlooked most of the time. Nowadays diagnostic methods have evolved in such a way that they are available for most of the population. Among the treatment schemes is the triple therapy, which is the most effective because it reaches 90% levels of microorganism eradication. For that reason we considered it of great importance doing a review of the most relevant things in literature about this bacteria and the diseases it is associated. PMID- 15794415 TI - [Immunomodulation by tacrolimus in atopic dermatitis]. AB - Atopic dermatitis is a common allergic disease, in which the treatment is extremely complex; even when several immunological abnormalities have been described in atopic dermatitis, the immune response to drugs remains unclear for both: conventional and unconventional therapies. The present review is centered on clinical efficacy and safety of tacrolimus, one of the immunomodulators proposed to treat atopic dermatitis. There are clinical evidences to support that tacrolimus have considerable impact on expression of inflammatory markers, despite of clinical assays could be necessary to demonstrate its profiles of toxicity and efficacy, during long-time periods. PMID- 15794417 TI - Spinal cord injury nursing and evidenced-based practice. AB - The importance of the dissemination of current data-based research findings cannot be said enough! Nurse researchers have an obligation to report their findings and potentially improve nursing practice and patient care outcomes. Conclusions drawn from the "evidence," whether good, bad, or indifferent, benefit nursing practice. Nurses need to know "what works," "what doesn't work," and the pluses and minuses of every finding. Reporting the "evidence" at national and international platforms can reach many health care providers. To reach the largest possible audience, however, research findings must be published in refereed journals. SCI Nursing is a refereed journal that reaches across our national boundary to other countries and the entire world. Publishing scholarly works is a responsibility of professional nurses! PMID- 15794416 TI - [Eosinophilic colitis. A report of two cases with non conventional treatment]. AB - Eosinophilic colitis is a rare entity of unknown etiology characterized by diarrhea, abdominal pain, and gastrointestinal bleeding. Diagnosis includes histopathological infiltration of more than 20 eosinophils in colon. It is frequently associated with milk hypersensitivity and, less usual, with other foods and increased IgE. Histopthological appearance of eosinophil mediators has been observed in the gut. It is sometimes related to the degree of infiltration of eosinophils in the gut as well as to the disease severity. There is not an established treatment for this entity, although systemic steroids have been used with certain efficacy. However, there is a recurrence of the symptoms when the therapy stops, besides the well known side effects of the long-term use of steroids. Cromolyn inhibits mast cell degranulation and prevents liberation of mediators. It is successful in certain cases, specially the severe ones. However, it is not available for its use in our country. Ketotifen, as last resource in our patients with bad response to habitual treatment and restriction diet, was used. Although its use is controversial, we consider that stabilizing mast cell membrane with subsequent inhibition of degranulation and recruitment of eosinophils to sites of inflammation, would also restrain histamine liberation and blockage of H1 receptors, which would diminish local damage induced by eosinophils. Nonetheless ketotifen mechanism of action is unknown, our patients improved after treatment with this drug. PMID- 15794418 TI - Creative innovations: the door is open for SCI nurses. PMID- 15794419 TI - Developing and implementing transdisciplinary rehabilitation competencies. AB - The Commission on Accreditation of Rehabilitation Facilities (CARF) requires that brain and spinal cord injury (SCI) specialty programs assure a base competency level for staff. At the authors' institution, senior staff members from all rehabilitation disciplines developed a Competency Fair based on a transdisciplinary approach to care. Each module contained reading material and a competency test. Some modules required a return demonstration. Topics included dysphagia, skin care, cognition, and bedside emergencies, among others. At a 4 day Competency Fair, 95 of the 110 staff members completed the training. Additional training was provided to those who failed the examinations. The training met CARF standards, and staff expressed a greater appreciation of others' roles and felt empowered to share their roles with others. Greater crossover of duties among disciplines, and a better understanding of the transdisciplinary approach to care, were apparent. Staff also reported improvements in patient care and an expansion of their knowledge base. PMID- 15794420 TI - Provider attitudes and beliefs about clinical practice guidelines. AB - The goals of clinical practice guidelines (CPG) are to improve the process and outcomes of health care, decrease practice variation, and optimize resource utilization. The objectives of this study were to (a) describe overall provider attitudes and beliefs about CPG, and (b) describe provider attitudes and acceptance of two specific spinal cord injury (SCI) CPG. A total of 152 health care providers responsible for implementation of the CPG at participating Veterans Health Administration (VHA) SCI sites responded to a survey (response rate of 35%). Overall, SCI care providers expressed positive attitudes towards CPG, including the two SCI guidelines included in this study. A comparison of responses revealed relatively few areas in which differences existed among SCI facilities and provider groups. Nurses represented the largest provider group participating in this survey and consistently expressed the most positive responses. In particular, nurses were more positive about guidelines, recognized the benefits of the guidelines, and were more willing to support the development of guidelines, compared to other providers in the study. The results of this study suggest that negative attitudes and beliefs about guidelines might be less of an obstacle to guideline implementation in VHA SCI Centers. Nurses are in a position to play a key role in their implementation. PMID- 15794421 TI - Autonomic dysreflexia in acute spinal cord injury: incidence, mechanisms, and management. PMID- 15794422 TI - Trends in qualitative nursing research. PMID- 15794423 TI - Empowering leadership through magnetism. PMID- 15794424 TI - School re-entry after pediatric spinal cord injury. PMID- 15794425 TI - American Association of Spinal Cord Injury Nurses standards of practice--revised 2003-2004. PMID- 15794426 TI - Are you "Information literate"? A quick assessment for the spinal cord injury nurse. PMID- 15794427 TI - Seizing opportunity. PMID- 15794428 TI - Competency in aging: do we have it? PMID- 15794429 TI - Key elements of bladder and Bowel management for children with spinal cord injuries. AB - Bladder and bowel management can be one of the most challenging aspects of daily life after spinal cord injury (SCI). Children are especially unique due to their ever-changing growth and developmental considerations. Continence is crucial for children who are still developing self-confidence and self-esteem. Proper bladder and bowel management can impact long-term health and well-being and can also affect quality of life. Providing education and implementation of bladder and bowel management programs are essential for children with SCI. This article will provide a general overview of common bladder and bowel management options for children with SCI. Age-appropriate expectations will be described, as well as patient and family educational strategies, to promote independence when implementing bladder and bowel programs. PMID- 15794430 TI - Adjustment to the process of grief following a spinal cord injury/dysfunction. PMID- 15794432 TI - Enhancing return to work: matching SCI clients with long-term vocational goals. PMID- 15794433 TI - Perspectives in nutrition: feeding through the physiologic response to injury. PMID- 15794434 TI - Aging: a new opportunity to publish in SCI nursing. AB - Publishing in SCI Nursing is a rewarding experience in terms of professional growth and contributing to the body of knowledge of SCI nurses. "Guidelines for Contributors" are on the back, inside cover of each issue of SCI Nursing and on the AASCIN Web site. Collaborate with nursing colleagues, or interdisciplinary team members, to begin your manuscript today. PMID- 15794436 TI - Discovering the ethnonursing research method. PMID- 15794435 TI - Societal trends impacting children: recreation implications for those with spinal cord injuries. PMID- 15794437 TI - Clinical nurse leader: another role--more confusion? PMID- 15794438 TI - Thinking about depression. PMID- 15794439 TI - Chronic stress, salivary cortisol response, interpersonal relatedness, and depression among community-dwelling survivors of traumatic brain injury. AB - Depression is a common mood disorder after traumatic brain injury (TBI). Largely, study of this phenomenon is theoretical and without biological measures. This explanatory study, guided by McEwen's allostasis model of stress, examined relationships among chronic stress, salivary cortisol profiles, post-injury depression, and interpersonal relatedness. Seventy-five participants, who were or had participated in outpatient brain injury rehabilitation therapies and experienced mild-to-moderate levels of brain injury, were recruited for this cross-sectional study. Salivary cortisol levels showed the usual patterns of circadian rhythmicity, and those with milder injuries had higher 8 am cortisol levels. Salivary cortisol values were not related to measures of chronic stress, interpersonal relatedness, or depression with two exceptions. The 8 am and noon mean values were significantly greater for those who reported more pre-injury childhood adversity, while the 8 pm cortisol mean level was associated with the frequency of pre-injury stressful life events. For this outpatient sample, salivary cortisol levels do not appear to be elevated after TBI or to lack circadian rhythmicity as previously reported. There may be some value in using this measure as a correlate with persons treated in specialized TBI clinics who report pre-injury chronic stress, but future studies are needed with TBI persons who were not treated in specialized clinics or were not taking medications known to influence the hypothalamic-pituitary-adrenal axis. PMID- 15794440 TI - Nursing grand rounds: multiple sclerosis. AB - Multiple sclerosis (MS) is a highly variable, unpredictable disease and one of the most life-altering diagnoses a person can receive. Because it usually strikes in the prime of life, frequently progresses to disability, and has no cure, MS can make a strong emotional impact--not only on those who suffer from it, but also the healthcare team. Because MS is such a complex, multifaceted disorder, nurses who care for people with MS are faced with numerous clinical challenges. Many of the challenges are unique to MS, demanding, and time-consuming. Well informed nurses are positioned to evaluate and explain the disease process, assist in the alleviation of symptoms, educate partners and families, and help improve quality of life. A case example can help nurses understand the real-life concerns of a person with MS. PMID- 15794441 TI - Husbands and wives living with multiple sclerosis. AB - Multiple sclerosis (MS) frequently is diagnosed in young adults. Coping with symptoms of MS is challenging not only for the person with the disease, but also for his or her spouse. The well spouse often assumes the caregiving role. The purpose of this qualitative research was to investigate the experiences of persons whose spouses have MS. Twelve people participated in a 2-hour focus group: 8 men and 4 women. The husbands were, on average, 50 years old, and the wives averaged 55 years old. The length of time since diagnosis ranged from 2 to 11 years for the husbands and from 3 to 13 years for the wives. The focus group discussions were audiotaped and transcribed verbatim. Participants talked freely. Four major themes emerged: caregiver roles, need for information, relationship changes, and barriers. Men attempted to protect their wives' energy, intervening for them. Wives encouraged independence in their husbands. Spouses need information about MS, complementary interventions, and support. They want increased public awareness of invisible symptoms and awareness in the workplace of continuing capabilities of persons with MS. Role reversals were challenging for the women who felt that "MS is the third person in a marriage." Spouses need help to maintain appropriate boundaries. Limitations of the study include the small, economically homogeneous sample and the single encounter with the subjects. A longitudinal intervention study is needed. PMID- 15794442 TI - Mild traumatic brain injury in psychiatric inpatients. AB - The purpose of this study was to compare psychiatric inpatients with and without mild traumatic brain injury (MTBI) on the variables of length of stay (LOS), number of psychiatric admissions, and change from admission to discharge on Global Assessment of Functioning (GAF). The study sample included 54 psychiatric inpatients, 18 to 65 years old, with a history of MTBI and 52 comparison psychiatric inpatients absent histories of MTBI, matched on age, sex and diagnostic category. LOS was longer for those with MTBI than for those without MTBI. This difference was statistically significant, after adjustment for matching. When adjusted for baseline GAF, this LOS difference remained statistically significant. The number of prior psychiatric admissions was 19% higher for participants with MTBI than for comparison participants, and when the LOS analysis was adjusted for this factor, it remained statistically significant. GAF change-from-baseline did not differ between the MTBI and comparison groups. Data from this study suggest that psychiatric inpatients with a history of MTBI may differ in frequency and length of hospitalization from otherwise similar inpatients without MTBI history. PMID- 15794443 TI - Pupil examination: validity and clinical utility of an automated pupillometer. AB - Pupillary size and reactivity have long been a critical component of the clinical assessment of patients with neurological disorders. The pupillary examination may provide critical information related to new or worsening intracranial pathology and facilitate prompt intervention to minimize further neuronal damage. With this in mind, intensive care nurses caring for neurologically impaired patients frequently must perform pupillary examinations in concert with assigning a Glasgow Coma Scale score. The purpose of this study was to test the accuracy and reliability of an automated pupillometer compared with the standard manual examination as a preliminary step in assessing the usefulness of automated pupillometry in the critical care setting. Twenty patients in the intensive care units of a teaching hospital were examined by two groups of three examiners using both the manual examination with a penlight or similar light source and a portable automated pupillometer capable of measuring pupil size and reaction. Measurements by a static pupillometer before and after each pupillary examination were used to determine the mean "true" size of the pupil. This study found that the automated pupillometer is more accurate and reliable than the manual examination in measuring pupil size and reactivity. For these reasons, such a device may be a beneficial addition in the clinical assessment of neurologically impaired patients. PMID- 15794444 TI - The meaning of living with traumatic brain injury in people with moderate or severe traumatic brain injury. AB - A traumatic brain injury (TBI) extensively affects the injured person's daily life. Research based on the perspectives of people with TBI can increase understanding of the challenges they face and the possibility of supporting them in managing their lives. The aim of this study was to elucidate the meaning of living with TBI as narrated by the people with moderate or severe TBI. The data were collected by means of qualitative research interviews with 12 participants who had lived with TBI for 4-13 years. A phenomenological hermeneutic method was used to interpret the transcribed interviews. The study showed that people with TBI had lost their way and struggled to achieve a new normalcy. Losing one's way included experiences of waking up to unknown, missing relationships and experiencing the body as an enemy. Participants' struggles to attain a new normalcy included searching for an explanation, recovering the self, wishing to be met with respect, and finding a new way of living. Living with TBI seems to mean living with a perpetually altered body that changed the whole life and caused deep suffering, where feelings of shame and dignity competed with each other. Participants seem to be quite alone in their suffering and need more support from healthcare professionals. PMID- 15794445 TI - Applying the psychoneuroimmunology framework to nursing research. AB - Psychoneuroimmunology (PNI) provides a distinct perspective regarding the interrelatedness of the nervous, endocrine, and immune systems. PNI explicates the possible means by which behavior and emotion can influence immune function. Moreover, PNI explains the means by which the immune system affects the nervous system and affects psychological response. The interactions among these systems are mediated at the molecular level by cytokines and hormones produced by cells of not just the immune but also the nervous and endocrine systems. These cytokines and hormones affect endocrine and neuronal processes that, in turn, affect mood, emotions, personal perception, as well as the immune response. Analysis of the effect of cytokines and hormones at the molecular, cellular, and peripheral level is under intense investigation. Such analysis will lead to a better understanding of the connections among the psychological, neurological, and immunological systems. This understanding will provide for a holistic perspective upon which better health care can be provided, discomfort minimized, and disease prevented. The use of such a perspective in neuroscience nursing investigation can elicit novel approaches to care. PMID- 15794446 TI - Morphometric study of the human muscle spindle. AB - OBJECTIVE: To study the morphometric characteristics of the human muscle spindle in normal muscle and to investigate the influence of aging. STUDY DESIGN: The following variables were studied in 72 spindles: area and diameter of the spindle; thickness of the capsule; number, area and diameter of fibers; and number and area of nuclei. RESULTS: In deltoid and extensor digitorum brevis muscles, a reduction in the diameter of the spindle as a function of age was found, while no statistically significant change in the variables as a function of age was observed in the quadriceps femoris and biceps muscles. In the deltoid, a reduction in the number of fibers and an increase in their diameter were also observed. CONCLUSION: These findings could prove useful in the study of the spindle in relation to disease. PMID- 15794447 TI - Assessment of NMYC amplification: a comparison of FISH, quantitative PCR monoplexing and traditional blotting methods used with formalin-fixed, paraffin embedded neuroblastomas. AB - OBJECTIVE: To determine the degree of agreement between fluorescence in situ hybridization (FISH), Southern blot analysis and LightCycler monoplex polymerase chain reaction (PCR) analysis in the assessment of NMYC gene amplification status in neuroblastoma. STUDY DESIGN: We performed a retrospective analysis of NMYC amplification, using FISH, LightCycler monoplex PCR and Southern blot techniques to assess NMYC amplification in a series of 18 neuroblastomas and 20 histologically normal tissues (15 lymph nodes, 2 pancreas specimens, 1 section each of thyroid, prostate and uterus). RESULTS: Nine neuroblastomas were NMYC amplified, and the remaining cases were nonamplified. All cases yielded interpretable results by Southern blotting and PCR monoplexing techniques. A single case of neuroblastoma was difficult to interpret by FISH due to high background debris. A single case demonstrated low-level NMYC amplification by LightCycler PCR monoplexing but was nonamplified by the other 2 techniques. FISH analysis in 1 case showed amplification, while the other 2 techniques demonstrated nonamplified status. The case in which FISH analysis incorrectly demonstrated amplification was the same one in which there was high background debris. The Southern blot results were reported as amplified or nonamplified, while numeric amplification ratios were obtained by both FISH and PCR LightCycler monoplex analysis. Comparison of these techniques demonstrated FISH to underestimate the degree of amplification in cases in which the amplification level was high by PCR. In fact, FISH appeared to saturate at amplification ratios > 10. CONCLUSION: The study revealed a high level of concordance between the 3 techniques for assessment of NMYC amplification status. However, FISH analysis has the advantage of allowing concurrent assessment of NMYC amplification status and architecture. LightCycler PCR monoplexing appears to have the advantage of more accurately quantitating high levels of NMYC amplification, including those amplified 20-fold or higher. Both FISH and PCR LightCycler monoplexing have the advantage of being performable on formalin-fixed, paraffin-embedded tissue. PMID- 15794448 TI - Second order discriminant analysis in chemopreventive efficacy measurement. AB - OBJECTIVE: To describe the use of second order discriminant analysis as a classification methodology along with the underlying assumptions and sampling requirements, with special emphasis on the use of this analysis in chemopreventive efficacy studies. STUDY DESIGN: The discriminant function score distributions derived in an analysis of 2 diagnostic groups may show such overlap that a statistically significant difference in mean values cannot be shown and, more important, that a useful case-based classification cannot be attained. By using the discriminant function score distributions from each case, it is frequently possible to derive a second order discriminant function based on case specific characteristics, rather than characteristics of nuclei, thereby attaining improved case classification. RESULTS: Second order discriminant analysis has proven very useful in the documentation of case-level efficacy in chemopreventive trials. In a study of orally administered vitamin A, a first order discriminant analysis did not achieve a statistically significant difference in the score distributions for nuclei, but a second order discriminant analysis allowed a correct recognition of intervention effects in 85% of submitted cases. In a chemopreventive study of triamcinolone, a similarly inadequate discrimination based on discriminant function scores for nuclei resulted. After a second order discriminant analysis, a reduction in solar actinic damage could be shown in 14/15, or 93%, of treated cases. CONCLUSION: Second order discriminant analysis can be highly effective when the discriminating information offered at the nuclear level is inadequate due to high dispersion and small differences in mean values of discriminant function scores for the diagnostic groups. Second order analysis utilizes case-specific characteristics of the discriminant function score distributions to document diagnostic group separation and/or efficacy of chemopreventive intervention by a reduction in case discriminant function scores. PMID- 15794449 TI - Urothelial changes induced by therapeutic procedures for bladder cancer. A review. AB - Topical and systemic therapeutic agents and treatment modalities, such as thiotepa and mitomycin C, cyclophosphamide, bacillus Calmette-Guerin, radiation therapy, photodynamic and laser therapy, and gene therapy, produce a host of changes in the bladder, some of them mimicking cancer. Pathologists must be aware that following these types of treatment, the clinical utility of urinary cytology is reduced. PMID- 15794450 TI - Potential of radial basis function neural networks in discriminating benign from malignant lesions of the lower urinary tract. AB - OBJECTIVE: To investigate the potential value of morphometry and neural network tools for discriminating benign from malignant nuclei and lesions of the lower urinary tract. STUDY DESIGN: The study group consisted of 33 cases of lithiasis, 41 cases of inflammation, 66 cases of benign hyperplasia of the prostate, 4 cases of carcinoma in situ, 48 cases of grade 1 transitional cell carcinoma of the bladder (TCCB) and 123 cases of grade 2 and 3 TCCB. Images of routinely processed voided urine smears stained by the Giemsa technique were analyzed by a custom image analysis system. Analysis of the images gave a data set of features from 31,158 nuclei. A radial basis function (RBF)-type neural network was employed to discriminate benign from malignant nuclei, based on the extracted morphometric and textural features. Subsequently a second RBF classifier was employed to discriminate benign from malignant cases. The nuclei from 156 randomly selected cases (50% of total cases) was used as a training set, and the nuclei from the remaining 159 cases made up the test set. Similarly, in an attempt to discriminate at the patient level, the same 156 cases were used to train an RBF classifier; the remaining 159 cases were used for the test set. The cases used for training and testing the 2 classifiers (nuclear and patient level) were the same for the 2 kinds of classifiers. RESULTS: Application of the RBF classifier permitted the correct classification of 93.64% of benign nuclei and 85.61% of malignant, giving an overall accuracy of 84.45%. At the patient level the RBF classifier permitted an overall accuracy of 94.97%. These results were on the test sets. CONCLUSION: The role of nuclear morphologic features in the cytologic diagnosis of lower urinary tract alterations was confirmed by the results of this study. The observed overlap in feature space indicates that the nuclear characteristics do not form strictly separate clusters; that fact explains the difficulty morphologists have with reproducible identification of nuclei from the lower urinary tract. Application of RBF offers good classification at the nuclear and patient level and promises to become a powerful tool for everyday practice in the cytologic laboratory. PMID- 15794451 TI - Color image analysis for quantifying renal tumor angiogenesis. AB - OBJECTIVE: To segment and quantify microvessels in renal tumor angiogenesis based on a color image analysis method and to improve the accuracy and reproducibility of quantifying microvessel density. STUDY DESIGN: The segmentation task was based on a supervised learning scheme. First, 12 color features (RGB, HSI, I1I2I3 and L*a*b*) were extracted from a training set. The feature selection procedure selected I2L*S features as the best color feature vector. Then we segmented microvessels using the discriminant function made using the minimum error rate classification rule of Bayesian decision theory. In the quantification step, after applying a connected component-labeling algorithm, microvessels with discontinuities were connected and touching microvessels separated. We tested the proposed method on 23 images. RESULTS: The results were evaluated by comparing them with manual quantification of the same images. The comparison revealed that our computerized microvessel counting correlated highly with manual counting by an expert (r = 0.95754). The association between the number of microvessels after the initial segmentation and manual quantification was also assessed using Pearson's correlation coefficient (r = 0.71187). The results indicate that our method is better than conventional computerized image analysis methods. CONCLUSION: Our method correlated highly with quantification by an expert and could become a way to improve the accuracy, feasibility and reproducibility of quantifying microvessel density. We anticipate that it will become a useful diagnostic tool for angiogenesis studies. PMID- 15794452 TI - DNA ploidy and immunomarking of bladder urothelial tumors before and after intravesical bacillus Calmette-Guerin treatment. AB - OBJECTIVE: To investigate DNA ploidy and immunoexpression of Ki-67 and p53 as predictivefactors in cases of superficial urothelial cell carcinoma (UCC) treated with bacillus Calmette-Guerin (BCG). STUDY DESIGN: Samples were obtained from 66 patients with UCC (pTa grade 3 or high grade and pT1 independent of grade or with concomitant carcinoma in situ) before and after intravesical BCG treatment. DNA ploidy analysis (ploidy balance, degree of hyperploidy and aneuploidy, proliferation index) was done by static cytometry. Ki-67 and p53 were analyzed immunohistochemically in paraffin-embedded tissue, and their quantification was carried out using an image analysis system. RESULTS: During a mean follow-up of 63.8 months, 31 of the 66 patients developed recurrent tumors (46.9%). DNA ploidy analysis showed that ploidy balance as well as degree of hyperploidy and aneuploidy were not statistically different between recurrent and nonrecurrent tumors. Only proliferation index was statistically significant between recurrent and nonrecurrent tumors. No statistically significant difference was observed in the percentage of Ki-67- and p53-positive cells between primary tumors that recurred and those that did not. CONCLUSION: These findings suggest that only proliferation index has predictive value for recurrence and progression in UCC treated with BCG. PMID- 15794453 TI - Short-term impact of a national dental education program on children's oral health and knowledge. AB - OBJECTIVE: The objective of this four-week, examiner-blind study was to determine the impact of an educational oral health program conducted within a Boys and Girls Club of America in Chicago, Illinois, USA. The educational program focused on the gingival health (gingivitis and plaque) of participating children who were between the ages of six and 15. METHODOLOGY: The multi-week program taught the participants the basics of oral biology and disease, as well as proper oral health prevention, including oral hygiene, dietary modification, and the importance of visiting the dentist. A calibrated examiner measured whole mouth Loe-Silness Gingival Index (GI) and Turesky Modification of Quigley-Hein Plaque Index (PI) at baseline (immediately prior to the initiation of the educational program) and four weeks later. The primary efficacy analysis was based on the change from baseline for 90 subjects who were enrolled at baseline, participated in the educational program, and were examined four weeks later. In addition, subjects completed five questions at baseline and at four weeks to assess their oral health knowledge before and after exposure. RESULTS: Mean baseline GI score was 0.184, while the four-week mean GI score was reduced to 0.140. This represents a 24% reduction in GI score, with p < 0.001. The mean baseline bleeding site score was 9.99, while the four-week mean bleeding score was reduced to 7.97. This represents a 20% reduction in bleeding sites, with p = 0.005. The mean baseline PI score was 3.06, while the four-week mean PI score was reduced to 2.97. This represents a 3% reduction in PI score with p = 0.044. The subject population was found to have statistically significantly (p < 0.001) greater knowledge following the program at week four, with 37% (33 of 90) and 69% (62 of 90) of subjects answering the questions correctly at baseline and week four, respectively. CONCLUSION: Collectively, these data support the role of an educational program in promoting improved oral health in these children over a four-week period. PMID- 15794454 TI - Use of a novel group of oral malodor measurements to evaluate an anti-oral malodor mouthrinse (TriOralTM) in humans. AB - OBJECTIVE: This study compared the ability of a test mouthwash containing zinc chloride and sodium chlorite (TriOral) to reduce intrinsic oral malodor, to that of two other mouthrinses, one with zinc chloride only and the other with no zinc chloride/no sodium chlorite, using a novel group of oral malodor parameter measurements. METHODOLOGY: Forty-eight subjects completed the study; 16 in the test group, 17 in the zinc only group, and 15 in the no zinc chloride/ no sodium chlorite group. At baseline and after two and four weeks, parameters assessed were 1) malodorants in the headspace of and in solution in resting whole saliva determined organoleptically, 2) breath volatile sulfur compounds (VSC) measured with a sulfide monitor (Halimeter), 3) fresh and incubated saliva oxidation reduction potential (E(h)) measured with a platinum electrode, and 4) level of saliva indolic compounds (IC), indole and skatole, determined colorimetrically with Kovac's reagent. The VSC, E(h), and IC data for the three mouthrinses were analyzed statistically by repeated measures ANOVA between groups, and by 2-way ANOVA within groups. Corresponding organoleptic data were analyzed by Kruskal Wallis and Friedman non-parametric tests. RESULTS: Organoleptic, VSC, and E(h) evaluations clearly showed the zinc chloride/sodium chlorite test mouthrinse to be more effective than the other two rinses. In all cases, the level of significance was p < 0.001 between the test mouthrinse and its no zinc chloride/no sodium chlorite control; between test mouthrinse and the zinc chloride only product, significance was p < 0.05, < 0.001 and < 0.01 for the organoleptic, VSC, and E(h) tests, respectively. Noteworthy was the observation that the mean organoleptic saliva headspace score with the test mouthrinse was reduced to zero, and VSC levels fell below 50 ppb S by the end of the study, a level where the breath is usually non-odorous. The test mouthwash also appeared more effective in reducing the salivary IC levels, but the results did not reach significance at p < 0.05 unless IC levels were amplified in the saliva by incubation overnight at 37 degrees C. Correlations between the various procedures were highly significant, achieving in almost all cases a probability level of p < 0.001. CONCLUSION: The results supported the conclusion that the zinc chloride plus sodium chlorite mouthrinse (TriOral) is more effective in reducing oral malodor than a zinc chloride alone mouthrinse, and even more effective than its no zinc chloride/no sodium chlorite mouthrinse control. The methods used in this study were consistent with one another, and highly effective in measuring various parameters that characterize oral malodor. PMID- 15794455 TI - Effects of hydrogen peroxide bleaching strip gels on dental restorative materials in vitro: surface microhardness and surface morphology. AB - OBJECTIVE: This study examined the effects of peroxide tooth bleaching, including Crest Whitestrips hydrogen peroxide gel treatments, on the surface hardness and morphology of common dental restorative treatments. METHODOLOGY: American Dental Association (ADA) recommended dental restorative materials, including amalgam, dental gold, porcelain, glass ionomer, and composites, were prepared according to manufacturers' instructions. A cycling treatment methodology was employed which alternated ex vivo human salivary exposures with bleaching treatments under conditions of controlled temperature and durations of treatment. Bleaching treatments included commercial Crest Whitestrips bleaching gels, which utilize hydrogen peroxide as the in situ bleaching source, and several commercial carbamide peroxide bleaching gels. Control treatments included placebo gels and an untreated group. Crest Whitestrips bleaching included treatment exposures simulating recommended clinical exposures (14 hours), along with excess bleaching simulating exposure to five times suggested Crest Whitestrips use. At the conclusion of treatments, surface microhardness measures and surface morphological assessments with standard and variable pressure (VP-) SEMs were conducted to assess the effects of bleaching exposure on the surface morphology and structural integrity of the restoratives. RESULTS: Surface microhardness and SEM measures revealed no significant deleterious effects on the restoration surfaces from Whitestrips gels. CONCLUSION: These results confirm that tooth bleaching from the selected commercial hydrogen peroxide or carbamide peroxide bleaching systems does not produce changes in surface morphology or microhardness of common dental restorative materials. These results support the clinical safety of the selected commercial bleaching systems to the oral environment, matching results obtained from long-term use of these ingredients applied in dental offices and available in commercial formulations. PMID- 15794456 TI - Clinical response of three whitening products having different peroxide delivery: comparison of tray, paint-on gel, and dentifrice. AB - OBJECTIVE: Comparative clinical research was conducted to evaluate the efficacy and safety of three peroxide-containing tooth whitening products having different peroxide delivery. METHODOLOGY: A total of 43 healthy adults who met entrance criteria were randomly assigned to either a dual-phase, anticavity, 1% hydrogen peroxide dentifrice with a manganese gluconate activator, an 18% carbamide peroxide paint-on gel, or a 5% carbamide peroxide professional custom tray system. Following manufacturer's instructions, the activated dentifrice was used at least twice daily for two minutes, while the paint-on gel was applied twice daily. The custom tray, a barrier system and the experimental control for this study, was worn continuously for six to eight hours daily. Tooth color (L*a*b*) was measured on the maxillary anterior teeth from standard digital images, while safety was assessed from examination and subject report. RESULTS: At Day 15, the custom tray group had a significant (p < 0.002) reduction in yellowness and increased lightness, with adjusted mean (SE) deltab* of -1.83 (0.210) and deltaL* of 1.45 (0.292). The custom tray group experienced on average greater color improvement compared to either the paint-on gel or activated peroxide whitening dentifrice, differing significantly (p < 0.01) from either of the barrier-free systems with respect to deltab*, deltaL*, deltaE*, and deltaW*. In comparison, 14 days' use of the paint-on gel and activated dentifrice did not result in significant (p > 0.10) color improvements from baseline for deltab*, deltaL*, or deltaW*, with these two barrier-free systems not differing significantly (p > 0.26) with respect to any individual or composite color parameters. Tooth sensitivity and oral irritation were the most common safety findings in the tray and dentifrice groups (there were no adverse events in the paint-on group), and no subject discontinued treatment early because of a treatment-related adverse event. CONCLUSION: In head-to-head 14-day testing, a low concentration (5% carbamide peroxide) barrier-based tray system yielded superior tooth color improvement compared to two barrier-free delivery systems-an 18% carbamide peroxide paint-on gel and a 1% hydrogen peroxide dentifrice with a metal activator. PMID- 15794457 TI - Placebo-controlled clinical trial evaluating a 10% hydrogen peroxide whitening strip. AB - OBJECTIVE: A placebo-controlled clinical trial was conducted to evaluate the effectiveness and safety of a 10% hydrogen peroxide strip-based whitening system in subjects with tooth discoloration and no previous history of tooth whitening. METHODOLOGY: Informed consent was obtained, and 39 healthy adults were randomly assigned to either 10% hydrogen peroxide whitening strips (Crest Whitestrips Premium) or placebo strips with no peroxide. Strips were used unsupervised twice daily for 30 minutes over a seven-day period. At day four and day eight, tooth color (L*a*b*) was measured on the maxillary anterior teeth from standard digital images, and safety was assessed from examinations and subject reports. RESULTS: After three treatment days, the 10% hydrogen peroxide strip group had significant (p < 0.0001) color improvement from baseline, with means (SD) for deltab* and deltaL* of -1.57 (0.472) and 1.72 (0.619), respectively. Continued strip use from day four to day eight resulted in significant (p < 0.002) incremental improvement in both yellowness and lightness. By the end of treatment (day eight), the adjusted mean (SE) color change between groups was -2.20 (0.275) for deltab* and 2.24 (0.254) for deltaL*, a highly significant (p < 0.0001) color improvement for the 10% hydrogen peroxide group relative to placebo, of similar absolute magnitude for deltab* and deltaL*. Treatment with the 10% hydrogen peroxide strips was well-tolerated, with tooth sensitivity and oral irritation representing the most common findings. CONCLUSION: In this double-blind, placebo controlled clinical trial, statistically significant tooth whitening was evident after three days' treatment with 10% hydrogen peroxide whitening strips, and color improved with continued usage over seven days. PMID- 15794459 TI - Be true to thyself as a public health professional. PMID- 15794458 TI - New laboratory methods to study tooth surface coverage and interproximal plaque control by dentifrice products. AB - OBJECTIVE: To develop and test an in vitro tooth model for use in conjunction with laboratory methods to study interproximal effects and efficacy of dentifrices. The application of the model should offer visual evaluation of dentifrice coverage of the tooth surface, and measure dental plaque control at posterior interdental spaces with a dentifrice. METHODOLOGY: The dentifrice products tested with the model were: Colgate Total 2 in 1 Toothpaste and Mouthwash (CTTM), Colgate Total dentifrice (CTD), and Colgate Regular dentifrice (CRD). Extracted human posterior teeth were disinfected, cleaned, aligned, and mounted in denture acrylic. In the area coverage method, tooth surface coverage and penetration of two different forms of dentifrice products (CTTM and CRD) were compared using digital photography. In the interproximal plaque control method, the teeth were coated with human saliva and incubated anaerobically with a mixture of representative oral bacteria for six hours at 37 degrees C. In vitro dental plaque was assessed after brushing the facial surface with one of the three dentifrice products using a clinical plaque scoring index. RESULTS: The area coverage method demonstrated that both dentifrice products tested covered approximately 70% of the facial tooth surface; the CTTM dentifrice coverage on the lingual tooth surface was significantly higher than the coverage for the CRD dentifrice. With the interproximal plaque control method, in the presence of an active ingredient, the CTTM dentifrice had equivalent efficacy to the CTD dentifrice. Both CTTM and CTD were significantly superior to the CRD for interproximal dental plaque control. CONCLUSION: Using the developed tooth model, two assessment methods have been shown to have the potential to demonstrate tooth surface coverage, and to assess the potential efficacy of a dentifrice for the control of interproximal dental plaque. This process can indicate potential clinical evaluation of an oral care product, and support clinical findings with controlled evidence. PMID- 15794460 TI - Analysis of Bacillus sphaericus in controlling mosquito populations in urban catch basins. AB - The West Nile virus (WNV) is a mosquito-borne infection that can be fatal to humans, equines, and avians, among others. WNV was first introduced to the United States in 1999 and is rapidly spreading across the country. Urban catch basins are thought to be prime breeding sites for mosquitoes, especially those identified as WNV carriers. The pilot study reported here was conducted in a city in southeastern Pennsylvania where there are 70,000 catch basins. The purpose of the study was 1) to determine whether catch basins are breeding sites for mosquitoes and 2) to test the effectiveness of a larvicide that uses a bacteria, Bacillus sphaericus, to eliminate mosquitoes in urban catch basins. The two pronged study determined that catch basins are ideal locations for mosquitoes, especially Culex pipiens, and that B. sphaericus is an effective larvicide. PMID- 15794461 TI - Impact of restaurant hygiene grade cards on foodborne-disease hospitalizations in Los Angeles County. AB - Although health departments routinely inspect restaurants to assess compliance with established hygienic standards, few data are available on the effectiveness of these efforts in preventing foodborne disease. The study reported here assessed the impact on foodborne-disease hospitalizations in Los Angeles County of a restaurant hygiene grading system that utilized publicly posted grade cards. The grading systm was introduced in January 1998. Hospital discharge data on foodborne-disease hospitalizations were analyzed for Los Angeles County and, as a control, for the rest of California during the period 1993-2000. Ordinary least squares regression analysis was done to measure the effect of the grading progam on these hospitalizations. After baseline temporal and geographic trends were adjusted for, the restaurant hygiene grading program was associated with a 13.1 percent decrease (p < .01) in the number of foodborne-disease hospitalizations in Los Angeles County in the year following implementation the program (1998). This decrease was sustained over the next two years (1999-2000). The results suggest that restaurant hygiene grading with public posting of results is an effective intervention for reducing the burden of foodborne disease. PMID- 15794462 TI - Indoor air pollutants: limited-resource households and child care facilities. AB - This paper reports on a study of indoor air quality in homes and child care facilities in non-metropolitan counties of New York State. Specific pollutants examined were lead, radon, carbon monoxide, asbestos, and mold. Some homes had high levels of pollutants, and certain pollutants were significantly and negatively correlated with household income. High levels of pollutants also were observed in many child care facilities, which raises questions about constant exposure of children to pollutants. Recommendations are made for lowering pollutant levels in low-income households and child care facilities. PMID- 15794463 TI - An assessment of drinking-water supplies on the Hanford site: an evaluation conducted at a federal nuclear facility in southeastern Washington state. AB - Drinking water is supplied to most U.S. Department of Energy (DOE) facilities on the Hanford Site by DOE-owned, contractor-operated pumping and distribution systems. Water is primarily obtained from the Columbia River, but some facilities use water from on-site groundwater wells. Because of the large amount of radioactive and chemical waste produced, stored, and disposed of at Hanford, some people are concerned that waste materials are contaminating on-site drinking water supplies. This paper describes the drinking-water facilities and treatment requirements on the Hanford Site and summarizes radiological and non-radiological water quality data obtained from water samples collected from each drinking-water system in use during 2001 and 2002. Monitoring data show that Hanford-produced radionuclides are measurable in some drinking-water samples. The only non radiological contaminants detected either were by-products of the chlorination process or came from off-site agricultural activities. Contaminant level values were, in all cases, below state and federal drinking-water limits. This information will provide assurance to current employees and future site developers that drinking water on the Hanford Site is safe for public consumption. PMID- 15794464 TI - Ribosomal DNA (rDNA) identification of the culturable bacterial flora on monetary coinage from 17 currencies. AB - The aim of the investigation reported in this paper was to identify the bacterial microflora on monetary coinage from 17 countries by employment of polymerase chain reaction (PCR) sequenced-based molecular identification of rDNA from bacterial cultures. Silver, bronze, and other alloy coins (approximately 300 g) from 17 currencies were enriched individually by aerobic culturing in tryptone soya broth for 72 hours at 30 degrees C. Next, 20 microL of broth was inoculated onto Columbia blood agar supplemented with 5 percent volume-pervolume (v/v) defibrinated horse blood for 72 hours at 30 degrees C, and resulting colonies were purified by further subculture, as detailed above, for a further 72 hours. All colonies were identified by initial PCR amplification of a partial region of the 16S rDNA gene locus, which was then sequenced, and the sequence was aligned according to the BLASTn algorithm. Twenty-five isolates were obtained from the coinage; of these, 25 (100 percent) were Gram positive, and the most prevalent genus observed was Bacillus (B. megaterium, B. lentus, B. litoralis, B. subtilis, B. circulans and other Bacillus spp.), which accounted for 10 of 25 isolates (40 percent) and was isolated from 10 of 17 countries (58.8 percent). It was followed in prevalence by Staphylococcus spp. (Staph. aureus, Staph. epidermidis, Staph. hominis, Staph. schleiferi), which accounted for 7 of 25 isolates (28 percent) and were isolated from 7 of 17 countries (41.2 percent). Given the organisms identified in this study, it is not believed that monetary coinage presents any particular risk to public health. The authors support the principles of basic hygiene, however, in terms of proper handwashing and the avoidance of handling money when working with food or dressing wounds and skin lesions, In conclusion, the study demonstrated that money from 17 countries was contaminated by environmental Gram-positive flora, in particular Bacillus spp., and that the universal 16S rDNA-PCR approach coupled with automated direct sequencing provides a rapid means of identifying the contaminant organisms present. PMID- 15794465 TI - K-12 school health and safety--where to start. PMID- 15794466 TI - A systems-based food safety evaluation: an experimental approach. PMID- 15794467 TI - Effect of calf thymus extract and zinc supplementation on the cellular response of mice exposed to restraint stress. AB - The studies were carried out on Balb/c mice exposed to restraint stress twice for 12 h at 24 h intervals. Prior to stress exposure, the mice were treated with calf thymus extract (TFX - Jelfa) i.p. at a dose of 10 mg/kg, ten times at 24 h intervals. TFX was used per se or with zinc ions interaction, by adding zinc ions (as sulfate salt) to drinking water at a dose of 72 microg/mouse per day. The results obtained show that restraint stress dramatically decreased the total number of thymocytes and splenocytes which is also accompanied by decreasing weight ratio of the thymus and spleen. The decreasing number of thymic and spleen cells corresponded to a diminishing percentage of immature, double-positive CD4+CD8+ thymocytes, mature single-positive CD4+ thymic cells and CD4+, CD8+ and CD19+ splenocytes. Changes in the number of thymic cells affect their activity, which is expressed as a decreased proliferative response of thymocytes stimulated in vitro with concanavalin A (Con A) and phytohaemagglutinin (PHA). Besides, exposure to the restraint stress decreased interleukin-1 (IL-1) production by murine intraperitoneal macrophages stimulated in vitro with lipopolisacharide (LPS) from E. coli. Previous treatment with TFX counteracted restraint stress induced immunosuppression, which is expressed as partial normalisation of the total number of thymic and spleen cells, accelerated regeneration of these two lymphatic organs, shortned suppressive action of restraint stress on the percentage of immature CD4+CD8+ thymocytes and CD4+ splenocytes and in total normalisation of the CD4+ thymocytes and CD8+ splenocytes. TFX administered prior to restraint stress not only counteracted the suppresive effects of stress on the proliferative activity of thymic cells stimulated in vitro with Con A and PHA, but also augmented the proliferative response of these cells to two mitogens. The immunorestorative effect of TFX was augmented by zinc supplementation. PMID- 15794468 TI - Evaluation of specific humoral immune response in pigs vaccinated intradermally with deleted Aujeszky's disease vaccine and challenged with virulent strain of Herpesvirus suis type 1. AB - A comparison of intradermal (ID) versus intramuscular (IM) routes of pig vaccination with deleted Aujeszky's disease (AD) vaccine on the formation of specific postvaccinal and postchallenge humoral immune response was performed. The studies were carried out on 21 eight week-old piglets, divided into three groups--two experimental and one control of 7 piglets each. Animals of first two groups were vaccinated twice in 12 and 16 week of age with deleted, live attenuated AD vaccine Porcilis Begonia (Intervet). Group I was vaccinated with a dose of 2.0 ml (10(6.0) TCID50)) intramuscularly (IM) into neck muscles, and group II received 0.2 ml (10(5.0) TCID50) intradermally (ID) in neck area using needleless apparatus SERENA model SD 1-2 (Emplast, Italy). In group K (control) 2.0 ml PBS IM was used. Seventy days after the first vaccination all pigs were intranasally infected with a dose of 10(5.5) TCID50 of virulent Northern Ireland Aujeszky-3 (NIA-3) strain of Herpesvirus suis type 1 (SHV-1) by instilling 0.5 ml of virus suspension into each nostril. Specific humoral immune response was evaluated using seroneutralization (SN) test and gE-ELISA-Pseudorabies virus gpI Antibody Test Kit (Herd Chek Anti-PRV gpI), IDEXX Lab Inc (USA). It was found that challenge caused anamnestic reaction in both groups of vaccinated pigs, but postchallenge immune response was stronger in ID-vaccinated group--on 14 day post infection (dpi) SN antibody level was considerably higher than in IM-vaccinated group. The obtained results suggest that secondary immunological response after challenge is decidedly more effective in the range of evaluated parameters in animals vaccinated by ID route, which can be linked to, perhaps underestimated yet and seldom utilized, skin immunity mechanisms in specific prophylaxis of infectious diseases. Advantages and disadvantages of SN test and ELISA are also discussed. PMID- 15794469 TI - Flow cytometric assay in peripheral blood of dogs--reference values for leukocytes from Brazilian beagles. AB - Use of domestic reference values in the flow cytometry analysis is known to improve its accuracy by integrating local variations as gender, race and age. Up to date application of flow cytometry in veterinary medicine has been limited to describe the percentual values just for peripheral lymphocytes subsets of blood. We now report establishment of reference values for a wide range of proportional and absolute numbers of peripheral blood leukocytes, including T cells subsets, B cells, monocytes and eosinophils, applicable to the healthy population of Beagles in Brazil and other regions with similar demographic characteristics. Normal reference values were also established to estimate the gender-related differences. This information will provide clinical aid in the evaluation of immunologic status as well as standard values for experimental animals of dogs from Brazil and other similar regions. PMID- 15794470 TI - Biological properties of Roakin strain of NDV and TK900 strain of ADV after serial passages in CECC in the presence of methisoprinol and KLP-602. AB - Twenty serial passages of the TK900 strain of Aujeszky's disease virus (ADV) and the Roakin strain of Newcastle disease virus (NDV) were made in a chicken embryo cell culture (CECC), in the presence of two antiviral agents: Methisoprinol and KLP-602. The physicochemical properties of passaged viruses were determined. The results obtained suggest that Methisoprinol causes changes in the structure of viral proteins, whereas KLP-602 affects the envelope-dependent properties of the virus. It was also found that the alternations observed in passaged viruses were temporary phenotypic changes only, and not a consequence of permanent transformations of their genotypes. PMID- 15794471 TI - The effect of Lactobacillus acidophilus and Bifidobacterium spp. administration on the morphology of the gastrointestinal tract, liver and pancreas in piglets. AB - The aim of the study was to investigate the influence of administration of probiotic bacteria on morphology of the gastrointestinal tract, liver and pancreas. The experiment was performed on 15 piglets at the age from 3 to 35 days, intragastrically administered with Bifidobacterium breve, B. animalis, and Lactobacillus acidophilus bacteria. In all piglets examined, the development of the gastrointestinal tract, liver and pancreas was observed to progress normally. After microflora administration to the piglets, an increase in the number of fibrocytes and fibroblasts was observed in the mucosa lamina propria of stomach and intestines. An increase was also reported in the number and activation of endocrine cells in the stomach and small intestines. The activity of alkaline and acid phosphatases, as well as succinic (SDH) and lactic (LDH) dehydrogenases, was found to be higher after the administration of probiotics. The administration of bacteria, especially of Lactobacillus acidophilus, caused an increase in the number of lymphocytes and lymphoid cells in lamina propria and intraepithelial lymphocytes in the small intestine. Enhanced proliferation of crypt cells was observed in the crypts of intestinal glands; however, there were no statistically significant differences in the PCNA index between the control and probiotic administered groups. The performed study showed that the administration of probiotic bacteria has no negative impact on the morphology of the gastrointestinal tract, liver and pancreas and is found beneficial to its functioning and immune processes. PMID- 15794472 TI - Occurrence of enterotoxigenic strains of Staphylococcus aureus in raw poultry meat. AB - The aim of this work was to determine the contamination of raw poultry meat with enterotoxigenic strains of S. aureus, using the PCR method. PCR is a rapid and sensitive method, which can show the presence in food of enterotoxigenic strains of S. aureus on the basis of specific gene sequences and detect the potential source of contamination before enterotoxins are produced. No coagulase-positive staphylococci strains were found in 65 samples of chicken parts, but these bacteria were present in 11 of 23 examined samples of minced turkey meat (48%). Using the primers for enterotoxin genes A to C, 4 of the 11 isolated S. aureus strains showed a positive result in the PCR. Three of the isolates represented the SEB gene and remaining one the SEC gene. The results obtained showed that PCR is sensitive and rapid method which may be used for detection and identification of enterotoxigenic S. aureus. PMID- 15794473 TI - Genotype analysis of Campylobacter jejuni isolated from broilers in Poland. AB - Twenty-six Campylobacter jejuni strains isolated from poultry were analyzed by genotypic typing including ITS-profiling, REP- and ERIC-PCR. ITS-profiling revealed the presence of 8 different genotypes. Amplification of REP sequences by PCR gave similar results with 10 different genotypes. ERIC-PCR was found to be the most discriminatory for typing C. jejuni. As many as 13 different DNA patterns were obtained with this technique. Based on data obtained it was found that C. jejuni isolates recovered from broilers at the slaughterhouses in southwest Poland are characterized by a high degree of genetic heterogeneity. PMID- 15794474 TI - Evaluation of different laboratory methods for diagnosis of pig chlamydiosis in Lithuania. AB - Pig chlamydiosis is antrophozoonosis caused by Chlamydophila abortus. Chlamydias (C type) are widely found in nature and can infect humans, domestic and wild mammals, and 139 types of birds. The peculiar feature of chlamydias is the tropism to different tissues, organs and organisms. In 2502 pig blood sera tests from Lithuanian farms, anti-chlamydia complement binding (CB) antibodies were detected in 192 cases (7.7%). Serological tests showed the following (C type) chlamydia bearing regions: 22.0% Mazeikiai district, 17.2%--Kaisiadorys district, 13.5%--Panevezys district, 12.3%--Vilkaviskis district. Rare incidence of the disease was found in Siauliai district 1,2% and Klaipeda district 2.5% farms. The highest antibody titers in blood serum tests were found in Joint Stock Company (JSC) "Krekenava" and "Vejine", i.e. 1:128 and 1:64, respectively. The following methods for the study of pig chlamydiosis were used and comparatively evaluated: complement binding reaction (CBR), direct immunofluorescence (DIF), imunoenzyme assay (IEA), indirect immunofluorescense (IIF), micro immunofluorescense (MIF), polymerase chain reaction (PCR) and cell culture (CC) test. PCR method was found to be more sensitive and reliable compared to imunoenzyme assay, but the latter is more economic especially for screaning. In pigs with the clinically expressed symptoms, 108 pigs infected with chlamydia were detected. CB assay revealed the infection rate from 3.4% to 7.9% in piglets, sows and boars. The highest level of chlamydia infection was detected in fatteners (17.6%). Seroepizootic study of pig chlamydiosis revealed the different infection rate in the animals investigated. The highest chlamydia infection risk is in winter (10.4%) and the lowest--in summer (2.8%). PMID- 15794476 TI - Immunohistochemical characterization of neurons in the porcine ciliary ganglion. AB - The present study was aimed at disclosing the chemical coding of nerve structures in the porcine ciliary ganglion (CG) using immunohistochemical methods. The substances under investigation included markers of "classical" neurotransmitters, choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DbetaH) as well as neuropeptides, somatostatin (SOM), galanin (GAL), substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY). Immunoreactivity to ChAT and VAChT was found virtually in all the neuronal somata and in numerous intraganglionic, varicose nerve fibres which often formed basket-like formations around the nerve cell bodies. Many CG neurons contained immunoreactivity for SOM (46%) or GAL (29%). Interestingly, a small number (approx. 1%) of the cholinergic somata stained for TH but not for DbetaH; nevertheless, some extra- and intraganglionic nerve fibres displayed immunoreactivity for DbetaH or TH. The CG perikarya stained neither for vasoactive intestinal polypeptide (VIP) nor for neuropeptide Y (NPY), but some NPY- or VIP-positive nerve terminals were observed within nerve bundles distributed outside the ganglion. SP- and CGRP-immunoreactivity was found in some intraganglionic nerve fibres only. The present study revealed that the porcine CG consists of cholinergic neurons many of which contain SOM and GAL. Thus, it can be assumed that in the pig, these neuropeptides are involved, complementary to acetylocholine, in the parasympathetic postganglionic nerve pathway to structures of the eye including the ciliary and iris sphincter muscles. PMID- 15794475 TI - The influence of male pheromones on the contractile reactivity of the isolated superficial veins of the nose and face during the estrous cycle in gilts. AB - This study was designed to determine whether steroid sex pheromones of the boar may affect the contractile activity of the superficial venous vessels of the nose and face in gilts, and in this way contribute to recently discovered humoral transfer of pheromones to the brain and hypophysis. The dependence between the reactivity of nasal and facial veins to male pheromones and the phase of the estrous cycle in gilts was also studied. The gilts were used in the luteal phase of the cycle and in the follicular phase of the cycle. The dorsal nasal, frontal and facial veins were isolated on an appropriate day of the estrous cycle. The isolated rings of veins were treated with androstenone (5alpha-androst-16-en-3 one), androstenol (5alpha-androst-16-en-3-ol) and testosterone (17beta-hydroxy-4 androsten-3-one) in concentrations of 1 or 10 microM. Changes in the contractile activity of the isolated vein segments were measured using isometric transducer and recorded on HSE-ACAD software for Windows. Androstenone--main signaling boar pheromone--induced much stronger reactions of the vessels than androstenol. Androstenone caused significant relaxation of the dorsal nasal and frontal veins, and significant increased tension of the facial vein in the follicular phase of the estrous cycle. The results obtained suggest a direct effect of male pheromones on the contractile reactivity of the superficial veins of the nose and face in the female, and in this way contribute to a humoral pathway for transfer of pheromones to the brain and hypophysis. Moreover, the present study revealed changes in the reactivity during the estrous cycle of the veins, transporting blood from the region of the nasal cavity, to male pheromones participating in the regulation of female reproductive processes. PMID- 15794477 TI - Immobilization (restraint) stress in desert sheep and goats, and the influence of pretreatment with xylazine or sodium betaine thereon. AB - Some behavioral, clinical, biochemical and haematological parameters were studied in Desert (Najdi) sheep and goats subjected to the acute and unavoidable stressful stimuli of immobilization. The effect of pretreatment with xylazine (n = 6) or sodium betaine (n = 6) on the responses of these animals has also been investigated. The immobilization stress resulted in increased vocalization and in variable and statistically insignificant increases in heart pulse and respiratory rates. In the two species, immobilization stress significantly elevated the plasma concentration of cortisol (from about 35.2 to about 83.8 mmol/L) and glucose (from 3.1 to 4.6 mmol/L), and decreased the concentration of magnesium (from 0.81 to 0.65 mmol/L). The endogenous thiocyanate concentration was unaffected. The immobilization stress also insignificantly decreased the haematocrit (PCV), and the number of lymphocytes, and increased the concentration of haemoglobin. Pretreatment of sheep and goats with xylazine at a dose of 0.01 mg/kg, via the intravenous route significantly alleviated the effects induced by the stressful stimulus. Pretreatment of the two species with sodium betaine (10 mg/kg/day for 3 days), however, produced variable and insignificant effects. There were no significant differences between sheep and goats in the responses to the immobilization stress, except in vocalization, which was more pronounced in sheep than in goats. PMID- 15794478 TI - Role of goats in epizootiology and epidemiology of Q fever. AB - Numerous data show that the epizootics of Q fever in goats can be particularly related to cases of this disease in humans. The aim of the study was to examine 98 goat serum samples from the farm where abortions, early parturition and parturition of weak goatlings were noted. The microaglutination method was used in this study. Serum dilution 1:8 was defined as a positive titre. The study revealed that 79.6% of serum samples were positive and numerous high titres suggested an acute form of infection. It could be supposed that Q fever was the reason of abortions in the herd. The results obtained point to the necessity of examinations for Q fever in goats, because of the possibility of infection in people who have a contact with these animals. Goats seem to be the animal species especially sensitive to C. burnetii infection. Q fever should be taken into consideration in the differential diagnosis of the goat diseases, when abortions occur. PMID- 15794479 TI - COX-2 inhibitors and cardiovascular toxicity: a class effect? PMID- 15794480 TI - Aggressive systemic mastocytosis: a case report and brief review of the literature. AB - Aggressive Systemic Mastocytosis (ASM), a rare subtype of Systemic Mastocytosis (SM), results from clonal proliferation and invasion of multiple organs by neoplastic mast cells. The clinical presentation varies, dependent on which organ systems are involved, and may take an indolent or rapidly fatal course. Several treatment strategies have been proposed. The most effective treatment, though not curative, is a combination of glucocorticoids and alpha-interferon 2b (IFN alpha). We present a case report of ASM to demonstrate the clinical presentation, laboratory findings, and therapeutic response to treatment. PMID- 15794481 TI - Sports medicine. PMID- 15794482 TI - Biodegradable poly(terephthalate-co-phosphate)s: synthesis, characterization and drug-release properties. AB - To develop biodegradable polymers with favorable physicochemical and biological properties, we have synthesized a series of poly(terephthalate-co-phosphate)s using a two-step poly-condensation. The diol 1,4-bis(2-hydroxyethyl) terephthalate was first reacted with ethylphosphorodichloridate (EOP), and then chain-extended with terephthaloyl chloride (TC). Incorporation of phosphate into the poly(ethylene terephthalate) backbone rendered the co-polymers soluble in chloroform and biodegradable, lowered the Tg, decreased the crystallinity and increased the hydrophilicity. With an EOP/TC molar feed ratio of 80: 20, the polymer exhibited good film-forming property, yielding at 86.6 +/- 1.6% elongation with an elastic modulus of 13.76 +/- 2.66 MPa. This polymer showed a favorable toxicity profile in vitro and good tissue biocompatibility in the muscular tissue of mice. In vitro the polymer lost 21% of mass in 21 days, but only 20% for up to 4 months in vivo. It showed no deterioration of properties after sterilization by gamma-irradiation at 2.5 Mrad on solid CO2. Release of FITC-BSA from the microspheres was diffusion-controlled and exceeded 80% completion in two days. Release of the hydrophobic cyclosporine-A from microspheres was however much more sustained and near zero-ordered, discharging 60% in 70 days. A limited structure-property relationship has been established for this co-polymer series. The co-polymers became more hydrolytically labile as the phosphate component (EOP) was increased and the side chains were switched from the ethoxy to the methoxy structure. Converting the methoxy group to a sodium salt further increased the degradation rate significantly. The chain rigidity as reflected in the Tg values of the co-polymers decreased according to the following diol structure in the backbone: ethylene glycol > 2-methylpropylene diol > 2,2-dimethylpropylene diol. The wide range of physicochemical properties obtainable from this co-polymer series should help the design of degradable biomaterials for specific biomedical applications. PMID- 15794483 TI - Poly(propylacrylic acid)-mediated serum stabilization of cationic lipoplexes. AB - The serum instability associated with cationic lipoplexes represents one of the major obstacles for the in vivo delivery of nonviral gene therapy vectors. Recently, we have shown that poly(propylacrylic acid) (PPAA), a pH-sensitive polyanionic polymer, can significantly improve the in vitro serum stability of DOTAP lipoplexes and enhance transfection (Cheung et al., Bioconjug. Chem. 12, 906 (2001)). We investigated this serum-stabilizing effect provided by PPAA using methods to identify the specific serum proteins that interact with DOTAP/DNA and DOTAP/DNA/PPAA lipoplexes and determined their modes of interaction with these lipoplexes. Studies showed that only low-density lipoprotein (LDL) caused significant decondensation of DNA from lipoplexes lacking PPAA, but that fully condensed DNA was retained within lipoplexes incorporating PPAA. Another major factor in the loss of transfection activity was due to the reduced cellular uptake of DOTAP lipoplexes upon exposure to serum, with bovine serum albumin (BSA) and high-density lipoprotein (HDL) acting as major contributors to this reduction in vector internalization. In contrast, lipoplexes containing PPAA maintained high levels of uptake into cells in the presence of these proteins. Transfection results generally concurred with the mechanistic studies, suggesting that maintaining effective cellular delivery of intact lipoplexes in the presence of serum proteins is important to retain high transfection efficiencies. These results indicate that the addition of PPAA as a ternary component in DOTAP lipoplexes can overcome some of the serum-related deficiencies encountered with these lipoplexes to provide efficient transfection. PMID- 15794484 TI - Immobilization of streptavidin-horseradish peroxidase onto a biotinylated poly(acrylic acid) backbone that had been radiation-grafted to a PTFE film. AB - Films of polytetrafluoroethylene (PTFE) were modified by radiation graft polymerization of acrylic acid (AAc). Optimal conditions for efficient AAc grafting were studied, including pre-irradiation dose in air, monomer concentration, temperature and time of the grafting process. Carboxylic groups of the grafted polyAAc were activated with carbodiimide (EDC) for biotinylation by reaction with 5-(biotinamido) pentylamine. Streptavidin-horseradish peroxidase (SA-HRP) was immobilized by affinity complexation of the SA with the biotin groups on the PTFE surface. The amount of active HRP immobilized on the PTFE films was determined as a function of the extent of polyAAc grafting. This study has demonstrated the utility of combining the processes of (a) radiation grafting of polymers with reactive groups onto inert polymers such as PTFE, (b) biotinylation of the graft polymer reactive groups, (c) immobilization of streptavidin on the biotinylated surface sites, followed by (d) immobilization of biotinylated, biologically active molecules via complexation of their conjugates with streptavidin. In this study, the last two steps were combined by immobilizing the complex of streptavidin and biotinylated HRP onto the biotinylated surface sites. The unique nature of this process is the ability to immobilize biotinylated molecules on an inert surface as PTFE. PMID- 15794485 TI - Semi-interpenetrating network of poly(ethylene glycol) and poly(D,L-lactide) for the controlled delivery of protein drugs. AB - We have prepared a semi-interpenetrating network (IPN) of poly(ethylene glycol) dimethacrylate (PEGDMA) with entrapped poly(D,L-lactide) (PLA) using photochemical techniques. These IPNs were developed for the controlled delivery of protein drugs such as growth factors. The PEG component draws water into the network, forming a hydrogel within the PLA matrix, controlling and facilitating release of the protein drug, while the PLA component both strengthens the PEG hydrogel and enhances the degradation and elimination of the network after the protein drug is released. The rate and extent of swelling and the resultant protein release kinetics could be controlled by varying the PEG/PLA ratio and total PLA content. These IPNs were prepared using a biocompatible benzyl benzoate/benzyl alcohol solvent system that yields a uniform, fine dispersion of the protein throughout the PEG/PLA IPN matrix. IPNs composed of high molecular mass PLA and lower PEG/PLA ratios exhibited lower equilibrium swelling ratios. The release of bovine serum albumin (BSA), a model protein, from these IPNs was characterized by a large initial burst, regardless of the PEG/PLA ratio, due to the entrapment of residual solvent within the network. Microparticles of the PEG/PLA IPNs were also prepared using a modified Prolease strategy. Residual solvent removal was significantly enhanced using this process. The microparticles also exhibited a significant reduction in the initial burst release of protein. Mixtures of different compositions of PEG/PLA microparticles should be useful for the delivery of a variety of protein drugs with different release kinetics from any tissue-engineering matrix. PMID- 15794486 TI - Characteristics of chemically cross-linked myosin gels. AB - Myosin gels, 10 mm x 10 mm x 1 mm in size, were obtained by chemical cross linking of scallop myosin using 1-ethyl-3-(3-dimethylaminoprolyl) carbodiimide hydrochloride (EDC), glutaraldehyde (GA), or transglutaminase (TG). All myosin gels showed high Mg-ATPase activity, although it sensitively depended on the species of cross-linker used. Among cross-linkers used, myosin gel cross-linked by TG showed the highest sensitivity, almost as high as that of native myosin. The motility assay of native actin filament on the myosin gel showed that all these myosin gels can give motion to actin filament. Among them the one cross linked by TG had the highest average velocity and the lowest threshold concentration of ATP for movement of the actin filament and the values are nearly the same as that of native myosin. In order to give the actin filament motion with preferential direction, we attempted to make myosin gel with oriented structure by applying a shear stress. Myosin gel with oriented filament array 1 cm long and 50 microm in diameter was obtained. We found that actin filaments prefer to move along the axis of the oriented myosin gel with an increased velocity. PMID- 15794487 TI - The relevance of the transfer of molecular information between natural and synthetic materials in the realisation of biomedical devices with enhanced properties. AB - Past and recent attempts to introduce in synthetic polymers molecular information from natural substances through simple blending, template polymerization and molecular imprinting are reviewed. The most promising approaches that can open the way to the realisation of new materials with improved biocompatibility, antibody- or enzyme-like performances are analysed more deeply. The realisation of bioartificial blends from natural and synthetic polymers, molecularly imprinted nanospheres or membranes that can act as recognition element in (bio)sensing devices, as synthetic enzymes or as key constituents of body fluids purification tools is presented in order to make the reader aware of the fascinating possibilities that these techniques make available to the biomedical science and engineering in the close future. The last part of the paper describes recent attempts to insert recognition elements for large molecules as proteins, DNA segments, viruses or whole cells in synthetic polymer systems, in order to develop new systems in the treatments of diseases and for tissue-engineering applications. PMID- 15794488 TI - Monocyte activation on polyelectrolyte multilayers. AB - The adherence and activation of primary human monocytes was investigated on a polyelectrolyte multilayer film containing hyaluronic acid (HA) and poly-L-lysine (PLL). The sequential layer-by-layer deposition of the multilayer film was characterized by surface plasmon resonance. Eight alternating bilayers displayed an effective thickness of 16.15 nm with a total polymer coverage of 2.10 microg/cm2. For cell studies, HA-PLL multilayers were constructed on tissue culture polystyrene (TCPS) substrates and characterized by time of flight second ion mass spectrometry (ToF-SIMS) analysis. Principal component analysis of the ToF-SIMS spectra resolved no significant difference in surface chemistry between PLL-terminated and HA-terminated multilayer surfaces. Monocyte adhesion on PLL- and HA-terminated surfaces was measured by the lactate dehydrogenase assay and showed a significant decrease in cell adhesion after 24 h incubation. Cell viability measured by Live/Dead fluorescent staining showed significant cell death in the adherent cell population over these 24 h. Tumor necrosis factor alpha (TNF-alpha) production, a measure of monocyte activation, was quantified by ELISA and normalized to the number of adherent monocytes. The activation of monocytes on PLL-terminated and HA-terminated surfaces was nearly identical, and both surfaces had TNF-alpha levels that were 8-fold higher than TCPS. These results demonstrate that sufficient PLL had diffused into the surface layer to direct monocyte adherence and to induce cytokine activation and cell death on the HA-terminated multilayer films. The diffusion of the second multilayer component to the coating surface should, thus, be taken into account in the design of polyelectrolyte-based biomaterial coating strategies. PMID- 15794489 TI - Enhancement of stability of glucose oxidase by immobilization onto metal ion chelated poly(N-vinyl imidazole) hydrogels. AB - Poly(N-vinylimidazole), PVIm, gels were prepared by gamma-irradiation polymerization of N-vinylimidazole in aqueous solutions as an affinity gel for glucose oxidase (GOx). These affinity gels with a water swelling ratio of 1800% for plain polymeric gel and between 30-80% for Cu(II) and Co(II)-chelated gels at pH 6.0 in phosphate buffer were used in the GOx adsorption studies. Maximum metal ion adsorption capacity of these hydrogels was found to be 3.64 mmol/g dry gel for Cu(II) and 1.72 mmol/g dry gel for Co(II) leading to GOx adsorption capacities of 343 and 528 mg enzyme/g dry gel, respectively, as compared to 228 mg for the plain dry PVIm gel. Activity studies were carried out using plain and the metal ion-chelated form of this hydrogel to investigate the stability and retained activity of the GOx in different buffer solutions and at different temperatures. Activity of the enzyme, either in free or immobilized form on the gel, decreased dramatically in acetate buffer solutions. In phosphate buffer solution, however, stability of enzyme has been found to be significantly high reaching 90% retained activity at the end of a 40-day period at 4 degrees C for Co(II) chelated systems. After immobilization of the enzyme onto metal-chelated hydrogel, the thermal stability of enzyme was enhanced significantly showing 23% activity, even at 75 degrees C. PMID- 15794490 TI - Synthesis of segmented poly(ether urethane)s and poly(ether urethane urea)s incorporating various side-chain or backbone functionalities. AB - A series of linear and branched functionalized high-molecular-weight segmented poly (ether urethane)s and poly(ether urethane urea)s were prepared by chain extending isocyanate pre-polymers based on poly(tetramethylene oxide) with molecular weight 1000 and 4,4'-diphenylmethane diisocyanate. Different functional groups were incorporated within the polymer backbone or on side-chains by using several chain extenders during the synthesis: glycerol, 2,2-bis (hydroxymethyl) propionic acid, 1H, 1H,2H,3H,3H-perfluoroundecane-1,2-diol, 1H,1H,8H,8H dodecanefluoro-1,8-octanediol, 1,3-diamino-2-hydroxypropane and 3,5 diaminobenzoic acid, using the method of gradual approach to stoichiometry. In some cases, pendant functional groups were used as reactive sites for the further attachment of side groups. Polymers were characterized using 1H-NMR and FT-IR spectroscopies and GPC in conjunction with chemical structural confirmation by a model compound comparison study of 4,4'-diphenylmethane diisocyanate or trifluoro p-tolyl isocyanate reacted with 1,3-diamino-2-hydroxypropane and 1,4-butanediol. PMID- 15794491 TI - Cancer survivorship and work. AB - This brief paper emphasizes that much of the knowledge and skills in the area of musculoskeletal disorders and work disability can be brought to bear on understanding and then intervening in the area cancer survivorship and work. PMID- 15794492 TI - Preliminary evaluation of the generalized work distress scale in patients with work-related upper extremity disorders. AB - This study evaluated the reliability and construct validity of the Generalized Work Distress Scale in 207 patients attending a specialty clinic with work related upper extremity disorders. Factor analyses were conducted and compared to the original developmental study confirming a two-factor solution: an internal factor related to Work-Related Dysphoria and an external factor related to Work Related Support. Reliability (internal consistency) of factors was high (Cronbach's alpha = 0.87 and 0.89). The Work-Related Dysphoria factor was more related to self-reported global depressed mood and self-reported upper extremity functional disability, than was Work-Related Support. This study provided preliminary support for the use of this scale in patients with work-related upper extremity disorders and consideration to depressive symptoms in these patients. Further investigation of the psychometric properties of this scale and its predictive validity are required. PMID- 15794493 TI - Management and employee agreement on reports of organizational policies and practices important in return to work following carpal tunnel surgery. AB - This study's purpose was to assess the agreement between management and employee ratings of organizational policies and practices (OPP) involved in the return to work process following carpal tunnel surgery. As a part of the prospective community-based Maine Carpal Tunnel II Study, 65 manager and employee pairs completed a questionnaire tapping four OPP dimensions. people oriented culture, safety climate, ergonomic practices, and disability management. It was hypothesized that managers and employees would agree on their assessment of the four OPPs and a composite organizational support index. Agreement was assessed using Lin's concordance correlation coefficient. Employee and manager ratings were similar for the organizational support index (rho(c) = 0.14, p = 0.08), and people oriented culture (rho(c) = 0.25, p = 0.01) but not the other three OPPs. In larger companies (>450 employees), ratings were also similar for safety climate (rho(c) = 0.24, p = 0.09), disability management (rho(c) = 0.22, p = 0.07) and ergonomic practices (rho(c) = 0.35, p = 0.02). In unionized companies there was agreement for safety climate (rho = 0.44, p = 0.02), disability management (rho(c) = 0.41, p = 0.01) and ergonomic practices (rho(c) = 0.40, p = 0.06). These preliminary results suggest employees can report on certain OPPs and that an employee questionnaire can be used to assess organizational support. Given recent evidence that employee ratings of OPPs are predictive of injury/illness incidence, work disability and return-to-work outcomes, further research is needed to confirm these findings. PMID- 15794494 TI - Predictors of upper extremity discomfort: a longitudinal study of industrial and clerical workers. AB - Upper extremity discomfort associated with work activity is common with a prevalence of over 50% in many settings. This study followed a cohort of 501 active workers for an average of 5.4 years. Cases were defined as workers who were asymptomatic or had a low discomfort score of 1 or 2 at baseline testing and went on to report a discomfort score of 4 or above on a 10-point visual analog scale. This change is considered clinically significant. Controls had a low baseline discomfort score and continued to have a low discomfort rating throughout the study. The risk factors found to have the highest predictive value for identifying a person who is likely to develop a significant upper extremity discomfort rating included age over 40, a BMI over 28, a complaint of baseline discomfort, the severity of the baseline discomfort rating and a job that had a high hand activity level (based upon hand repetition and force). The risk profile identified both ergonomic and personal health factors as risks and both factors may be amenable to prevention strategies. PMID- 15794495 TI - A longitudinal study of industrial and clerical workers: predictors of upper extremity tendonitis. AB - Upper extremity tendonitis (UET) associated with work activity is common but the true incidence and risk factors can best be determined by a prospective cohort study. This study followed a cohort of 501 active workers for an average of 5.4 years. Incident cases were defined as workers who were asymptomatic at baseline testing and had no prior history of UET and went on to be diagnosed with an UET during the follow-up period or at the follow-up evaluation. The incident cases were compared to the subset of the cohort who also had no history of an UET and did not develop tendonitis during the study. The cumulative incidence in this cohort was 24.3% or 4.5% annually. The factors found to have the highest predictive value for identifying a person who is likely to develop an UET in the near future included age over 40, a BMI over 30, a complaint at baseline of a shoulder or neck discomfort, a history of CTS and a job with a higher shoulder posture rating. The risk profile identifies both ergonomic and personal health factors as risks and both categories of factors may be amenable to prevention strategies. PMID- 15794496 TI - A longitudinal study of industrial and clerical workers: incidence of carpal tunnel syndrome and assessment of risk factors. AB - This study followed workers over an extended period of time to identify factors which may influence the onset of Carpal Tunnel Syndrome (CTS). The purpose was to evaluate incidence of CTS and to create a predictive model of factors that play a role in the development of CTS. This prospective study followed 432 industrial and clerical workers over 5.4 years. Incident cases were defined as workers who had no prior history of CTS at baseline testing and were diagnosed with CTS during the follow-up period or at the follow-up screening. On the basis of logistic regression, significant predictors for CTS include baseline median-ulnar peak latency difference, a history of wrist/hand/finger tendonitis, a history of numbness, tingling, burning, and/or pain in the hand, and work above the action level of the peak force and hand activity level threshold limit value. This longitudinal study supports findings from previous cross-sectional studies identifying both work related ergonomic stressors and physical factors as independent risk factors for CTS. PMID- 15794497 TI - A cross-sectional assessment of the ACGIH TLV for hand activity level. AB - The ACGIH Worldwide Threshold Limit Value (TLV) for hand activity "considers average hand activity level or "HAL" and peak hand force." We report cross sectional data that assess the validity of the TLV with respect to symptoms and selected upper extremity musculoskeletal disorders among workers. The prevalence of symptoms and specific disorders were examined among 908 workers from 7 different job sites in relation to the TLV. Worker exposures were categorized as above the TLV, above the TLV Action Limit but below the TLV, or below the TLV Action Limit. Symptoms in the distal upper extremities did not vary by TLV category. Tendonitis in the wrist/hands/fingers did not vary by TLV category, but elbow/forearm tendonitis was significantly associated with TLV category. All measures of carpal tunnel syndrome were associated with TLV category. In all instances, prevalence of symptoms and specific disorders were substantial in jobs that were below the TLV action limit, suggesting that even at "acceptable" levels of hand activity, many workers will still experience symptoms and/or upper extremity musculoskeletal disorders, which may be important in the rehabilitation and return to work of injured workers. Future analyses need to examine the incidence of symptoms and upper extremity musculoskeletal disorders prospectively among workers in relation to the TLV for hand activity. PMID- 15794499 TI - Darbepoetin alfa in anemia management: a vital role in cancer care. PMID- 15794498 TI - Measures of physical and cognitive function and work status among individuals with multiple sclerosis: a review of the literature. AB - The purpose of this review was to critically evaluate the multiple sclerosis (MS) literature that has examined physical and cognitive function in relation to ability to work. Although numerous factors may be considered when determining work ability, physical and/or cognitive functional limitations associated with MS are presumably the primary determinants of work capacity. An exhaustive search of the literature produced 20 research articles that described 18 studies. Findings from these studies support that limitations in physical or cognitive function can hinder one's ability to work; however, ability to work could not be based solely on these measures of function. Work ability among individuals extended beyond measures of impairment to include level of education, job characteristics, and disease symptoms such as fatigue. In summary, measures of physical and cognitive function can guide physicians when clinically evaluating an individual with MS, but are poor indicators for precluding an individual from working. PMID- 15794500 TI - Advances in the use of colony-stimulating factors for chemotherapy-induced neutropenia. PMID- 15794501 TI - Oral mucositis therapy comes of age. PMID- 15794502 TI - [Health preservation for workers is one of the most important objectives of health services]. AB - Based on profound analysis of medical supplies, morbidity, work conditions and work safety during this period of economic reforms in Russia, the authors justified priority directions, goals and tasks of health care system in protection and improvement of workers' health state. PMID- 15794503 TI - [Features of cardiologic diseases prevalence among individuals engaged into geology]. AB - The authors studied influence of work and living conditions on geologists' health state. A cohort under study demonstrated paradoxical epidemiologic situation with high mortality risk of arterial hypertension, whereas prevalence of this disease among specialists working on expeditionary shifted mode is nearly equal to that among general population. Unfavorable situation concerning cardiovascular diseases could be caused by high prevalence of cardiovascular risks and some occupational hazards. The situation is deteriorated by insufficient medical observation and psychologic traits of the specialists working on expeditionary shifted mode. PMID- 15794505 TI - [Work conditions and occupational features of female fur farmers]. AB - The authors present some problems faced during use of "R 2.2.755-99" Manual in evaluating work conditions of female fur farmers. Gender approach appears necessary in evaluation of work conditions through physical, chemical, biologic factors and work intensity. The authors suggest evaluation scale for work conditions in influence of negative odorants on workers. PMID- 15794504 TI - [Influence of occupational factors on immune state of workers producing building materials]. AB - The authors studied immune state of workers producing building materials, in accordance with the production features. Occupational hazards were proved to change contents and ratio of immune lymphocytes, to suppress the workers' nonspecific resistance. PMID- 15794506 TI - [Rehabilitating patients with occupational diseases on Siberian resorts]. AB - The authors present results of rehabilitation for vibration disease and respiratory diseases patients in sanatoriums and resorts. Complex special program caused considerable effects on serum peroxidant activity, immunologic state, microcirculation state in patients with vibration disease of I-II severity grade. Due to treatment, patients withoccupational pulmonary diseases demonstrated better general feeling and improved external respiration parameters. The authors showed that existing treatment strategies should be included into rehabilitation programs for patients with vibration disease and chronic pulmonary diseases. PMID- 15794507 TI - [Studying traits of dermatoglyphic signs distribution in vibration disease]. AB - The article contains results of study that covers 343 males exposed to occupational vibration in Kemerovo region. The authors studied features of dermatoglyphic signs distribution in vibration disease patients and in reference group, calculated parameters of relative risk for the disease, etiologic fraction and preventive fraction. Dermatoglyphic signs were proved to be informative markers for vibration disease risks. PMID- 15794508 TI - [Medical strategy in municipal institutions for workers' health care management]. AB - The authors stress increasing role of doctor in municipal medical institutions, with occupational medical service limited nowadays. The article contains typical medical defects in diagnosing occupational diseases and in their medical management. Suggestions cover measures to better those medical activities. PMID- 15794509 TI - [Cardiovascular diseases in workers of gas-processing plant]. AB - Considering prospective observation among gas-processing plant workers, the authors evaluated epidemiologic parameters associated with cardiovascular diseases. Findings are that functional cardiovascular disorders (vascular neurosis) appeared the most prevalent entity. Occupational environment could he considered as a risk factor for vascular neurosis in workers of gas-processing enterprise. PMID- 15794510 TI - [Reproductive function in stewardesses of civil airlines]. PMID- 15794512 TI - Beyond the horizon. PMID- 15794511 TI - [Health of workers engaged into general mechanical rubber goods production]. PMID- 15794513 TI - Taking back the ER. PMID- 15794514 TI - Clues to caring. PMID- 15794515 TI - Childbirth choices. A new study examines what hospitals need to know before reinventing birth. PMID- 15794516 TI - High expectations. PMID- 15794517 TI - What nurses want. PMID- 15794518 TI - Being direct. PMID- 15794519 TI - Here to stay? PMID- 15794520 TI - E-health prognosis. PMID- 15794521 TI - Welcome wagon. PMID- 15794522 TI - What's the buzz? PMID- 15794523 TI - Physiologic monitoring systems. AB - Physiologic monitoring systems monitor vital physiologic parameters so that clinicians can be informed of changes in a patient's condition. For this study, we evaluated systems from six monitoring suppliers--Drager Medical, GE Healthcare, Nihon Kohden, Philips Medical Systems, Spacelabs Medical, and Welch Allyn. The intent of this study is to help facilities choose not just the most appropriate system, but also the most appropriate version of that system--the combination of components that will best suit the facility's needs. Our testing focused primarily on adaptability, alarm implementation, and human factors design. We rated the systems based on their capabilities and performance for each of seven care settings: critical care unit, emergency department, intermediate care unit and general medical/surgical floor, operating room (with separate ratings for use during conscious sedation and general anesthesia), postanesthesia care unit, and transport. The systems performed well against the majority of our criteria. Nevertheless, we found notable differences in specific features and performance areas. These differences will have varying levels of significance for different hospitals. PMID- 15794524 TI - Rainout from Fisher & Paykel's 850 humidification system shuts down Respironics Esprit and adversely affects other ventilators. PMID- 15794525 TI - Understanding alarm recordings on Philips and other central station monitors. PMID- 15794528 TI - Independence, dignity, & choice in assisted living: interview with Patricia Polansky by Karen Amann Talerico. PMID- 15794529 TI - Mothers living with suicidal adolescents: a phenomenological study of their experience. AB - Adolescents who exhibit suicidal behaviors have a devastating effect on their mothers, but unfortunately, the mothers' experience is often the hidden dimension in the family. Currently, there is a considerable lack of research into the maternal experience of living with a suicidal adolescent. This phenomenological study describes and enhances the understanding of what life is like for 6 mothers living with suicidal adolescents. The data were collected through unstructured interviews. Thematic analysis using van Manen's methodology identified six themes: failure as a good mother, the ultimate rejection, feeling alone in the struggle, helplessness and powerlessness in the struggle, cautious parenting, and keeping an emotional distance. From these themes, the essence, "multiple loss and unresolved grief," was captured. The findings have implications for nursing education, practice, and research. In the provision of quality family-centered care, nurses can be educated and guided in their interventions by phenomenological research that recognizes the importance of the maternal perspective in the experience of living with suicidal adolescents. PMID- 15794530 TI - Risk assessment of forensic patients: nurses' role. AB - One of the unique roles of forensic nurses is to conduct risk assessments. Establishing a therapeutic nurse-patient relationship helps forensic nurses perform accurate and useful risk assessments. Accurate risk assessments can facilitate formulation of individualized risk management plans, designed to meet patients' needs and ensure public safety. The importance of forensic nurses' knowledge and application of appropriate communication and proper documentation cannot be overemphasized. PMID- 15794531 TI - The subpoena and a day in court: guidelines for nurses. AB - Psychiatric nurses or therapists may be subpoenaed to appear as direct or expert witnesses. The response must be carefully prepared, and nurses must be aware that testifying in court can be an anxiety-provoking experience. Testifying often involves being an advocate for a client. Clients who have a mental illness, are developmentally disabled, or have a history of abuse or recent violence deserve a conscientious response. The legal process is the avenue for providing safety or empowerment, for assuring that needs are met, and rights are obtained, and being part of that process is an important role. Being prepared for the role is essential. PMID- 15794532 TI - An unexpected hero. PMID- 15794533 TI - Correcting the medical literature: ethics and policy. PMID- 15794534 TI - Ossiculoplasty in a patient with a cleft of the soft palate. PMID- 15794535 TI - Bilaterally exposed carotid arteries after drainage of a sphenoid sinus mucocele. PMID- 15794536 TI - Glottic web at the striking zone. PMID- 15794537 TI - Vestibular findings in a 30-year-old woman with disabling dizziness following childbirth. PMID- 15794538 TI - Treatment of globus by upper esophageal sphincter injection with botulinum A toxin. PMID- 15794539 TI - Teratoma. PMID- 15794540 TI - Aggressive fibromatosis of the neck. PMID- 15794541 TI - Cystic hygroma exacerbated by pregnancy. PMID- 15794542 TI - 'Silent PPOs' and 'repricing' companies. PMID- 15794543 TI - Falling sensation in patients who undergo the Epley maneuver: a retrospective study. AB - The author conducted a retrospective study to determine the prevalence of a falling sensation in patients who underwent the Epley canalith repositioning maneuver for the treatment of benign paroxysmal positional vertigo. The author studied a total of 436 maneuvers performed on 412 patients and observed 58 episodes (13%) of a strong falling sensation, some very severe. In almost every case, the sensation occurred when the patient was moved to the final (sitting) position; in 1 case, the sensation occurred nearly 30 minutes later. The author recommends that physicians who perform the Epley maneuver warn patients of the risk of a falling sensation, take steps to prevent its consequences, and monitor their patients for at least 30 minutes after the completion of the procedure. PMID- 15794544 TI - The use of hydroxyapatite cement and a pericranial/deep temporal fascia graft for cranioplastic reconstruction of translabyrinthine craniectomy defects. AB - Most translabyrinthine temporal bone defects are reconstructed with free abdominal fat grafts, with or without the use of hydroxyapatite cement. However, these procedures are associated with considerable morbidity at the graft donor site, with a 6 to 15% incidence of cerebrospinal fluid (CSF) leaks, and with postoperative headaches. We have developed a new technique for reconstructive cranioplasty that involves the use of hydroxyapatite cement and a pericranial/deep temporal fascia graft. This technique obviates the need for an abdominal fat graft and therefore circumvents the morbidity associated with it; it may also significantly reduce the incidence of CSF leaks and postoperative headaches. We describe the results of our use of this technique in a series of 10 patients. Based on our early findings, we believe that this technique holds great promise for reconstructive cranioplasty following translabyrinthine craniectomy. PMID- 15794545 TI - Fibromatosis presenting as acute mastoiditis: a case report. AB - We describe the case of a middle-aged man who presented with manifestations of acute mastoiditis caused by fibromatosis of the mastoid region. A lesion of the right mastoid bone had eroded its wall and extended toward the middle and posterior cranial fossae. The macroscopic and microscopic appearance of an excised portion of the lesion established the diagnosis of mastoid fibromatosis. After a more detailed work-up, a second procedure involving extensive removal of the tumor was performed, and the diagnosis was confirmed. The patient's postoperative period was uneventful, and he showed no evidence of recurrence during 3 years of follow-up. PMID- 15794546 TI - Sinonasal hemangiopericytomas: clinicopathologic and imaging findings. AB - Hemangiopericytomas are rare soft-tissue neoplastic lesions that can arise in any part of the body. They are mesenchymal tumors that account for 3 to 5% of all soft-tissue sarcomas and 1% of all vascular tumors. They originate in extravascular cells (pericytes). Some 15 to 30% of all hemangiopericytomas occur in the head and neck; of these, approximately 5% occur in the sinonasal area. We describe our brief retrospective review of 7 histologically proven cases of sinonasal hemangiopericytoma, and we discuss the imaging characteristics and clinical and pathologic findings in these patients. PMID- 15794547 TI - The use of combined PET/CT for localizing recurrent head and neck cancer: the Pittsburgh experience. AB - We performed a retrospective study of 47 patients to ascertain the ability of combined positron-emission tomography and computed tomography (PET/CT) to localize recurrent head and neck cancer. When clinically warranted, biopsies were performed in an attempt to obtain pathologic confirmation of the PET/CT findings. Of the 47 patients, 33 exhibited PET/CT findings consistent with recurrent cancer. Of the 33 patients, 25 underwent either biopsy or surgical excision of disease in an attempt to obtain a pathologic confirmation. Biopsy analysis confirmed the PET/CT findings in 22 of these patients; in the remaining 3 patients, pathologic findings were inconsistent with the PET/CT diagnosis. Based on the subset of 25 patients who underwent pathologic testing, the sensitivity of combined PET/CT was 95% and the specificity was 60%. We conclude that combined PET/CT imaging is a valuable tool for localizing tumor recurrence in patients with head and neck cancer. PMID- 15794548 TI - Dynamic ultrasound-assisted extraction of colistin from feeds with on-line pre column derivatization and liquid chromatography-fluorimetric detection. AB - A dynamic ultrasound-assisted extraction (UAE) method with on-line pre-column derivatization/high performance liquid chromatography (HPLC) and fluorimetric detection is proposed for the analysis of colistin in feed. A flow injection manifold is used for the development of the extraction and derivatization steps and for interfacing them with the separation/detection step, thus providing an on line approach with the advantage of minimum sample handling. The derivatization was performed with ortho-phthaldialdehyde and 2-mercaptoethanol. The optimum conditions for colistin extraction and formation of the fluorescent derivative have been obtained by experimental design methodology. The use of a high intensity probe sonication makes UAE an expeditious (7 min versus > 1 h) and efficient (93.1-98.2% versus 87.5-94% of recovery) alternative as compared with extraction using an ultrasonic bath. The within-laboratory reproducibility and repeatability, expressed as percentage of relative standard deviation, were 5.2 and 5.8, respectively. PMID- 15794549 TI - Improved ultrasonic extraction procedure for the determination of polycyclic aromatic hydrocarbons in sediments. AB - The aim of this work was to optimize an ultrasonic extraction procedure for the determination of polycyclic aromatic hydrocarbons (PAHs) in sediments and to compare it with the reflux procedure using methanolic potassium hydroxide. Sample extracts were purified with a miniaturized silica gel chromatographic column and analyzed by gas chromatography-mass spectrometry (GC-MS). Ultrasonication using n hexane-acetone (1:1, v/v) solvent mixture on dried homogenized marine sediment gave better precision (smaller relative standard deviation (RSD) values) and comparable quantities of individual PAH's compared to the reflux procedure. Ultrasonication with the n-hexane-acetone (1:1, v/v) mixture, utilizing four 15 min extraction cycles, was found to be sufficient for extracting PAHs from wet sediments. The optimized ultrasonic extraction procedure extracted aliphatic and aromatic hydrocarbons from the National Institute of Standards and Technology SRM 1941a with recoveries greater than 90%. The major advantages of ultrasonication compared to the reflux method are the lower extraction times, simplicity of the apparatus and extraction procedure. The optimized ultrasonication procedure has been used in our laboratory to extract hydrocarbons from naturally wet sediments from rivers, and coastal and marine areas. PMID- 15794550 TI - Cationic surfactant-based polyfluorate salts: phase separation and analytical applications in the extraction and preconcentration of ionic species prior to liquid chromatography. AB - The liquid-solid phase separation originating from the formation of cationic surfactant-based polyfluorate salts (CSBPS) has been explored for extracting and preconcentrating ionic species. Two cationic surfactants were tested; one with aliphatic hydrocarbon tail [Cetyltrimethylammonium bromide (CTAB)]and the other containing a heterocyclic ring [Hexadecylpyridinium bromide (HPyBr)]. Phase separation possibility was investigated with the use of hexafluorophosphates (PF6 ) and tetrafluoroborates (BF4-). The effect of added acid, base and salt on the phase separation and analyte extraction was also investigated. In all cases the obtained phase diagrams consisted of two regions: a homogeneous liquid region and a solid-liquid region. Analytes of hydrophilic and hydrophobic nature such as amines, amino acids and organic chromophores were used as test compounds in both their anionic and cationic forms. The respective recoveries ranged from over 90% for anionic species and in the proximity of 50% for cationic species, remaining below 20% for neutral species. Extracts from alkaline aqueous and plasma samples spiked with tyrosine and phenylalanine were also subjected to HPLC separation with UV detection with satisfactory results. On line application was also enabled using a flow through-solid phase extraction-HPLC hyphenated apparatus, thus adding the element of automatization and increased reproducibility. PMID- 15794551 TI - Disposable ionic liquid coating for headspace solid-phase microextraction of benzene, toluene, ethylbenzene, and xylenes in paints followed by gas chromatography-flame ionization detection. AB - Solid-phase microextraction (SPME) with a disposable ionic liquid (IL) coating was developed for headspace extraction of benzene, toluene, ethylbenzene, and xylenes (BTEX) in paints. The SPME fiber was coated with IL prior to every extraction, then the analytes were extracted and desorbed on the injection port of gas chromatography, and finally the IL coating on the fiber was washed out with solvents. The coating and washing out of IL from the fiber can be finished in a few minutes. This disposable IL-coated fiber was applied to determine BTEX in water-soluble paints with results in good agreement with that obtained by using commercially available SPME fibers. For all the four studied paints samples, the benzene contents were under the detection limits, but relatively high contents of toluene, ethylbenzene and xylenes (56-271 microg g(-1)) were detected with spiked recoveries in the range of 70-114%. Compared to the widely used commercially available SPME fibers, this proposed disposable IL-coated fiber has much lower cost per determination, comparable reproducibility (RSD < 11%), and no carryover between each determination. Considering that IL possess good extractability for various organic compounds and metals ions, and that task specific IL can be designed and synthesized for selective extraction of target analytes, this disposable IL coating SPME might has great potential in sample preparation. PMID- 15794552 TI - Analytical methodology for the identification of estrogenic contaminants in fish bile. AB - Effluents from wastewater treatment works (WwTWs) contain estrogenic contaminants that can cause feminised responses in fish. In order to assess the identity of estrogenic contaminants taken up by fish exposed to effluents, an analytical method was developed to detect estrogenic substances in fish bile, where many xenobiotics are excreted and concentrated. Estrogenic metabolites in bile were deconjugated using enzymatic hydrolysis and the estrogenic activity was determined using a yeast estrogen receptor transcription screen (YES). Hydrolysed samples were concentrated by solid-phase extraction (SPE) prior to fractionation by reversed-phase high-performance liquid chromatography (HPLC). Active HPLC fractions were detected by YES assay and analysed by gas chromatography-mass spectrometry (GC-MS) after trimethylsilylation. The method was validated using bile samples from immature female rainbow trout, which had been exposed to either tap water or an undiluted estrogenic effluent for 10 days. Hydrolysis of bile from effluent-exposed fish was complete within 16 h add most of the estrogenic activity in the bile was released by 3-glucuronidase rather than sulfatase or 3 glucosidase treatment. The estrogenic activity of hydrolysed bile from effluent exposed fish ranged between 530 and 1440 ng E2eq/mL and was 17-48-fold greater than the activity of bile from reference fish exposed to tap water. The estrogenic activity of bile samples decreased with time in storage (at-70 degrees C by 7% per month). The recovery of estrogenic activity from SPE was 96 +/- 7% (mean +/- SD), from HPLC fractionation 87 +/- 7% and for the whole method 81 +/- 7% (n = 7). 17beta-Estradiol, estrone, 17alpha-ethinylestradiol, nonylphenol and short-chain nonylphenol polyethoxylates were all identified from GC-MS analysis of active HPLC fractions of bile from effluent-exposed trout, whereas only 17beta estradiol was detected in bile from fish exposed to tap water. There were also several other minor estrogenic components, at present unidentified, in bile of effluent-exposed fish. The work shows that fractionation of fish bile is a useful approach to identifying mixtures of estrogenic contaminants taken up by fish from WwTW effluents and has the potential for application in the detection of other endocrine disrupting chemicals in fish tissues. PMID- 15794554 TI - Two-dimensional liquid chromatography with mixed mode stationary phases. AB - Mixed mode stationary phases with ion-pairing reagent (acidic or basic) as integral part of hydrophobic chain offers unique selectivity, and hence, are ideal for multidimensional separations. The retention of hydrophobic components is a function of organic content, whereas that of charged species is a function of organic content, ionogenic modifier and its level in the mobile phase. Hence, by controlling the parameters influencing component retention (stationary phase and mobile phase), the selectivity of chemical components in the two-dimensional plane can be manipulated to improve the separation. A two-dimensional liquid chromatograph has been developed by coupling similar and dissimilar mixed mode stationary phases in the two dimensions. This technique has immense potential in resolving co-eluting components as the retention mechanism in the two-dimensions are complementary. However, with only part of the primary column eluent sampled into the secondary column, the technique is limited to qualitative analysis. PMID- 15794553 TI - High-performance liquid chromatographic assay of N(G)-monomethyl-L-arginine, N(G),N(G)-dimethyl-L-arginine, and N(G),N(G)'-dimethyl-L-arginine using 4-fluoro 7-nitro-2, 1,3-benzoxadiazole as a fluorescent reagent. AB - N(G)-Monomethyl-L-arginine (L-NMMA), N(G),N(G)-dimethyl-L-arginine (ADMA), and N(G),N(G)'-dimethyl-L-arginine (SDMA) are emerging cardiovascular risk factors. A high-performance liquid chromatographic method with fluorescence detection for the simultaneous determination of L-NMMA, ADMA and SDMA is described. The assay employed 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) as a fluorescent derivatization reagent. After solid phase extraction with cation-exchange column, the methylated arginines were converted to fluorescent derivatives with NBD-F, and the derivatives were separated within 32 min on a reversed-phase column. Nomega-Propyl-L-arginine was Used as an internal standard. Extrapolated detection limits were 12 nM (12 fmol per injection) for L-NMMA and 20 nM (20 fmol per injection) for ADMA and SDMA, respectively, with a signal-to-noise ratio of 3. The calibration curves for L-NMMA, ADMA and SDMA were linear within the range of 50-5000 fmol. The method was applied to the quantitative determination of L-NMMA, ADMA and SDMA in 200 microl of rat plasma. The concentrations of L-NMMA, ADMA and SDMA in rat plasma were 0.16 +/- 0.03, 0.80 +/- 0.25 and 0.40 +/- 0.21 microM, respectively (n = 5). PMID- 15794555 TI - Chromatographic evaluation of poly (trans-1,2-cyclohexanediyl-bis acrylamide) as a chiral stationary phase for HPLC. AB - Chiral stationary phases (CSPs) based on polymeric (R,R)- or (S,S)-1,2 diaminocyclohexane (DACH) derivatives are synthesized. When bonded to 5 microm porous spherical silica gel, the poly (trans-1,2-cyclohexanediyl-bis acrylamide) based poly-cyclic amine polymer (P-CAP) stationary phases is proved to be effective chiral stationary phases that could be used in the normal-phase mode, polar organic mode and with halogenated solvents mobile phases, if desired. Since these are entirely synthetic CSPs, the elution order of all enantiomers can be reversed between the (R,R) P-CAP and (S,S) P-CAP columns. Because of the high loading of chiral selectors, the columns exhibit very high sample capacities. Thus, P-CAP columns are useful for preparative and semi-preparative enantiomeric separations. The application of these CSPs and optimization of their separations are discussed. PMID- 15794556 TI - Evaluation of sugar sorption isotherm measurement by frontal analysis under industrial processing conditions. AB - This paper evaluates frontal analysis for routine sugar isotherm measurements at industrial conditions, that is concentrations up to 400 kg/m3 and a temperature of 60 degrees C. Sugar isotherms for a gel type cation-exchange resin loaded with metal ions were measured in a HPLC setup equipped with a UV detector. It is shown experimentally that isotherms obtained with large concentration steps (step series method) underestimated the isotherm. The underestimation is larger for larger resin particle size. In contrast, isotherms obtained with small concentration steps (staircase method) yielded correct isotherms. The seldom mentioned change of the sorbent volume during the course of an isotherm measurement is discussed. It is shown that shrinking of 4% cross-linked resin at high sugar concentration has a negligible effect on the isotherm. Furthermore, the isotherms obtained with staircase frontal analysis agreed very well with those obtained with the independent, though more laborious and time-consuming, adsorption-desorption method. Staircase frontal analysis is shown to be convenient and accurate and is therefore recommended for isotherm measurements covering large concentration ranges. PMID- 15794557 TI - A theory of protein-resin interaction in hydrophobic interaction chromatography. AB - Docking simulations were performed in order to investigate surface area of interaction between several ribonucleases and a reduced model for the hydrophobic moiety used in Phenyl Sepharose using the program AutoDock 3.0. For each ribonucelase, 80 independent simulations with populations consisting of 100 random structures were performed and from these the most probable docked protein ligand conformations were obtained. A new methodology was used to select the most probable conformations, based on qualitative and quantitative considerations. The interacting amino acids in each protein were identified. The average surface hydrophobicity of the interfacial zone (local hydrophobicity, LH) was determined. The LH showed a high correlation level (r2 = 0.99) with the "hydrophobic contact area" (HCA) experimentally determined for the different ribonucleases as well as with the dimensionless retention time (r2 = 0.90). This study allowed us to identify the zones on the protein surface most probably involved in protein retention in HIC, without tedious experimental work. Given the good correlation level obtained, this new methodology may constitute a novel approach that could be used to predict protein behavior in HIC. PMID- 15794558 TI - Simultaneous determination of sildenafil, vardenafil and tadalafil as forbidden components in natural dietary supplements for male sexual potency by high performance liquid chromatography-electrospray ionization mass spectrometry. AB - A high-performance liquid chromatographic method coupled with ultraviolet detection and electrospray ionization mass spectrometry (HPLC-UV-ESI-MS) was developed for simultaneous determination of banned additives-sildenafil, vardenafil and tadalafil in dietary supplements for male sexual potency. The separation was achieved on a C18 column with acetonitrile and aqueous solution (20 mmol ammonium acetate, 0.2% formic acid) as mobile phase at a flow rate of I ml/min with a linear gradient program. UV detection was at 292 nm. Identification of drugs was accomplished using ESI-MS. Good linearity between response (peak area) and concentration was found over a concentration range of 0.8-80 microg/ml for sildenafil; 2.25-225 microg/ml for vardenafil; and 1.1-110 microg/ml for tadalafil, with regression coefficient is better than 0.999. The recovery of the method ranged from 93.3 to 106.1%, and the relative standard deviation varied from 2.0 to 5.6% (n = 6). The method has been successfully applied to the analysis of practical samples of natural dietary supplements. PMID- 15794559 TI - Development, optimization and validation of a fingerprint of Ginkgo biloba extracts by high-performance liquid chromatography. AB - A four-step development, optimization and validation strategy for high performance liquid chromatography (HPLC) fingerprints of Ginkgo biloba extract is described. A suitable chromatographic system was selected first. The following step was performing a screening design to select important parameters. After selecting some controllable parameters and their range to further optimize, gradient optimization with uniform design was done. At last, method validation including determination of injection precision, repeatability, and a sample stability test, was performed. Through this effective and integrated four-step method, a feasible and reliable HPLC fingerprint to identify and assess the Ginkgo biloba quality can easily be established using a linear gradient elution with acetonitrile/0.1% phosphoric acid (from 14/86 to 30/70, v/v, in 40 min) as mobile phase, a column temperature of 30 degrees C and a detection wavelength of 350 nm. The strategy can also be applied for the development of fingerprints in the quality control of other herbal medicines. PMID- 15794560 TI - Liquid chromatographic method for quantifying polyphenols in ciders by direct injection. AB - An analytical method for the quantitative determination of the principal phenolic compounds (benzoic acids, hydroxycinnamic acids, 3-phenylpropionic acids, flavanols, procyanidins, dihydrochalcones, quercetin glycosides) in ciders, which successfully employs a RP-HPLC and photodiode-array detection system without prior treatment of the sample, is described. Parameters usually examined in the method validation were evaluated. Good linearity was obtained with correlation coefficients exceeding 0.999 and the detection limits ranged from 0.07 mg/L (p hydroxybenzoic acid) to 2 mg/L (hydrocaffeic acid). Recoveries ranging between 90 and 104% and the reproducibility of the method was always < 8% (RSD). The method was applied to a set of commercial samples and the results obtained may be helpful to establish a phenolic profile in Asturian cider. PMID- 15794561 TI - Rapid identification of RoHS-relevant flame retardants from polymer housings by ultrasonic extraction and RP-HPLC/UV. AB - A rapid method was developed for the extraction and identification of RoHS relevant organic flame retardants in polymer material. Extraction was carried out using ultrasonic supported solvent extraction (USSE) and isopropanol. HPLC separation was achieved using a reversed-phase phenylhexyl-modified column and methanol, containing 2-aminoethanol-buffered, alkaline water. Identification was carried out by scanning UV detection and comparison with a library assembled from spectra of reference substances. The method was used to extract and identify polymer additives in TV and PC monitor housings. The overall runtime required for extraction and chromatographic analysis is less than 10 min. The limits of detection comply with the recommendations set by the German draft law. PMID- 15794562 TI - Fluorescence labeling method for aryl halides with 4-(4,5-diphenyl-1H-imidazol-2 yl)phenylboronic acid based on Suzuki coupling reaction. AB - For the first time, fluorescence labeling methods for aryl halides with a fluorescent arylboronic acid was developed on the basis of a Suzuki coupling reaction. 4-(4,5-diphenyl-lH-imidazol-2-yl)phenylboronic acid (DPA) was used as a fluorescence labeling reagent. In order to explore its analytical performance, the reaction conditions were optimized using simple bromobenzene derivatives. The reactivity was then investigated with chloro- and iodobenzene derivatives, and also bromobenzene derivatives with different position of substituents. The order of reactivity with DPA: iodobenzene > bromobenzene more more than chlorobenzene derivatives, and p- > m- > o-substituted bromobenzenes. The detection limits of bromobenzene, 4-bromotoluene, and 4-bromoanisole ranged from 0.2 to 1.4 pmol/injection at a signal-to-noise ratio, (S/N) of 3. The applicability of the method to biological samples was also evaluated using clofibrate as the analyte. The reaction was found not only to proceed well but also to be selective for clofibrate even in the presence of plasma components. The method allowed the sensitive detection of clofibrate in human plasma with the detection limit of 170 pmol/mL (260 fmol/injection) at a S/N = 3. The proposed method is highly selective and sensitive and thus would be useful for labeling of aryl halides that do not have other functional groups that could be labeled by currently available fluorescent labeling reagents. PMID- 15794563 TI - High performance liquid chromatography equipped with a cathodic detector and column-switching device as a high-throughput method for a phosphatase assay with p-nitrophenyl phosphate. AB - LC coupled to an electrochemical detector (LC-EC) operating in cathodic mode with a column-switching system realizes a high-throughput detection of p-nitrophenol (NP). The measurement-time for each NP sample was shortened to 20 s, and the successive analyses of 39 samples was completed within 13 min. In the present system, the limits of detection and quantification were 0.15 and 0.20 microM, respectively, and further, up to 25 microM, a linear calibration curve was afforded. Relative standard deviations for standard solutions of 0.20, 1.0, and 25 microM NP were 4.3, 2.0, and 1.1% (n = 5), respectively. Between-run precisions of the analysis of 5.0 and 25 microM NP over 6 days were 4.8 and 1.3%, respectively. A comparison with the commonly used Bessey-Lowry-Brock method indicates that the present LC-EC is useful for the high-throughput assay of acid and alkaline phosphatases in urine and blood samples with a p-nitrophenyl phosphate substrate. PMID- 15794564 TI - A pulsed potential waveform displaying enhanced detection capabilities towards sulfur-containing compounds at a gold working electrode. AB - Pulsed electrochemical detection of sulfur-containing compounds was successfully investigated by applying a four-step potential waveform at a gold working electrode. This potential waveform called APAD, which stands for activated pulsed amperometric detection, is composed of an activation potential step added to a conventional three-step potential waveform. A key advantage of the APAD at the Au electrode is the ability to enhance sensitivity through the use of a short potential pulse (E(ACT) = +750 mV versus Ag/AgCl and tACT approximately 90 ms) during which the formation of redox active species (presumably OH*) are able to efficiently oxidize organosulfur compounds. The APAD waveform parameters were optimized to maximize the signal-to-noise ratio (S/N) and successfully applied for the sensitive detection of lipoic acid, biotin, iminobiotin, methionine, cystine, cysteine, homocysteine, homocystine, N-acetylcysteine and glutathione, following their separations by high-performance anion-exchange chromatography (HPAEC) using alkaline mobile phases. The detection limits (S/N = 3, 10 microL injected) ranged from 0.3 for cysteine (400 pg) to 0.02 micromol/L for biotin (50 pg) and methionine (30 pg). The response of sulfur-, amine- and alcohol-based compounds was compared by using four selected pulsed potential waveforms. It was found that the APAD exhibits excellent sensitivity for thiocompounds outperforming all other pulsed potential waveforms. Ratios of the peak areas for APAD and the six-step potential integrated waveform increased from 3.2 +/- 0.4 to 13.5 +/- 0.6 for lipoic acid and biotin, respectively. PMID- 15794566 TI - Volatile sulfur compounds in Cheddar cheese determined by headspace solid-phase microextraction and gas chromatograph-pulsed flame photometric detection. AB - The aim of this study was to develop a methodology for the analysis of volatile sulfur compounds (VSCs) in Cheddar cheese. Solid-phase microextraction (SPME) was employed to extract VSCs from the cheese matrix using a CAR-PDMS fiber. This extraction method was combined with gas chromatography-pulsed flame photometric detection (GC-PFPD) to achieve high sensitivity for sulfur compounds. The impact of extraction parameters, including time, temperature and sample size, was evaluated to determine the best conditions to analyze sulfur compounds in Cheddar cheese. Hydrogen sulfide, methanethiol, and dimethyl sulfide were found to constitute the majority of the overall sulfur profile while dimethyl disulfide and dimethyl trisulfide were present in lesser amounts. Artifact formation of volatile sulfur compounds was found to be minimal. Two commercial cheese samples were analyzed and differences in sulfur content were observed. Overall, SPME-GC PFPD was found to be a highly sensitive technique for the analysis of sulfur compounds in Cheddar cheese. PMID- 15794565 TI - Determination by gas chromatography/mass spectrometry of p-phenylenediamine in hair dyes after conversion to an imine derivative. AB - In this paper we describe an analytical method for the determination of p phenylenediamine (PPDA) in hair dyes. In the adopted methodology the analyte is transformed into the corresponding imine derivative by treatment with benzaldehyde, and then analyzed by gas chromatography (GC) combined to mass spectrometry (MS), operating in SIM conditions. The direct and simultaneous chemical derivatization of the two amino functions of the analyte with benzaldehyde enhances the instrumental responses enabling the use of a sensitive and accurate method. Concentrations of PPDA in a set of commercial hair coloring creams are determined making use of N-benzylidene-4-methylbenzene-amine as a very stable internal standard which is easily prepared by condensation of 4 methylbenzene-amine with benzaldehyde. The regression calibration curves for PPDA in hair dyes are linear within 0.1 +/- 25 mg/ml with 0.99 as a typical correlation coefficient. PMID- 15794567 TI - Investigation of methyl tert-butyl ether levels in river-, ground-, and sewage waters analyzed using a purge-and-trap interfaced to a gas chromatograph-mass spectrometer. AB - River water collected from 27 sites in 21 rivers, groundwater from 66 sites in 34 cities, and 2 sewage plants in the Niigata Prefecture, Japan (total 301 samples) were analyzed for methyl tert-butyl ether (MTBE) using an improved purge-and-trap GC-MS method. The amount of MTBE found in water samples from the 27 river sites ranged from 0.003 to 5.3 microg l(-1). MTBE was found in all 27 sites during winter but it was found at only 9 sites and 14 sites, respectively, during the summer. MTBE was found most frequently (in 21 sites) at levels ranging from 0.003 to 0.009 microg l(-1) during February. The highest levels of MTBE were always found in the samples collected from the same site. The amount of MTBE found in sewage water samples ranged from <0.003 to 0.025 microgl(-1). No significant differences were observed between the amounts of MTBE recovered from inflow and outflow waters. Relatively high levels (0.02-0.034 microg l(-1) ) of MTBE were found in January at two sites, which were located on the upper course of the Shinano River. MTBE levels ranged from 0.004 to 0.035 microgl(-l) and from 0.005 to 0.041 microgl(-1) at the mouths of the Shinano River and Agano River, respectively. The levels of MTBE in groundwater collected from 66 sites in 34 cities in Niigata Prefecture ranged from <0.003 to 5.9 microg l(-1). PMID- 15794568 TI - Determination of polydimethylsiloxane-seawater distribution coefficients for polychlorinated biphenyls and chlorinated pesticides by solid-phase microextraction and gas chromatography-mass spectrometry. AB - Applications of solid-phase microextraction (SPME) in the measurement of very hydrophobic organic compounds (VHOCs) are limited, partly due to the difficulty of calibrating SPME fibers for VHOCs. This study used a static SPME strategy with a large sample volume (1.6 L) and a five-point calibration procedure to determine the distribution coefficients for a large suite of polychlorinated biphenyls (PCBs) and chlorinated pesticides between a polydimethylsiloxane (PDMS) phase (100 microm thickness) coated on a glass fiber and seawater. An extraction time of 12 days was deemed adequate for equilibrium calibration from kinetic experiments. Two groups of randomly selected fibers divided into three batches (up to nine fibers in each batch) were processed separately with two gas chromatography-mass spectrometry (GC-MS) systems. Matrix effects arising from losses of the analytes to glass container walls and stirring bars were corrected. Relative standard deviations within the same batch were generally smaller than those for the entire group. Furthermore, KfVf (Kf and Vf are the distribution coefficient of an analyte between the polymer-coated fiber and aqueous phase and the fiber volume, respectively) values determined with two GC-MS systems were statistically different. These results indicate the calibrated KfVf values were less affected by the random selection of SPME fibers than by other experimental conditions, and therefore average KfVf values may be used for the same type of commercially available SPME fibers. The relative accuracy of our calibration method was similar to that of a previous study [P. Mayer. W.H.J. Vaes, J.L.M. Hermens, Anal. Chem. 72 (2000) 459] employing different coating thickness and calibration procedure. The present study also obtained a bell-shaped relationship between log Kf and log Kow (octanol-water partition coefficient) for PCB congeners with the maximum log Kf corresponding to log Kow approximately 6.5. This bell-shaped relationship was attributed mainly to steric effects arising from the interplay between the PDMS thickness and molecular sizes of the target analytes. PMID- 15794569 TI - Use of solid-phase microextraction/gas chromatography-electron capture detection for the determination of energetic chemicals in marine samples. AB - Gas chromatography with electron capture detection (GC-ECD) is a highly explosive sensitive analytical technique. However, its application to the analysis of sediment extracts is hampered by the presence of numerous endogenous interferences. In the present study, solid-phase microextraction (SPME) was used both as a purification technique for sediment extracts and as an extraction technique for water samples prior to analysis by GC-ECD. SPME/GC-ECD coupling was optimized and applied to the trace analysis of nine explosives including nitroaromatics and RDX in real seawater and marine sediment samples. Addition of a high concentration of salt (30%, w/v) in the aqueous medium and use of a carbowax/divinylbenzene (CW/DVB) coating led to optimal extraction efficiencies. Method detection limits (MDLs) ranged from 0.05 to 0.81 microg/L in water and from 1 to 9 microg/kg in dry sediment. Except for RDX, spike recoveries in seawater were satisfactory (89-147%) when samples were fortified at 2 microg/L of each analyte. Spike recoveries from dry sediment fortified at 10 microg/kg of each analyte gave lower recoveries but these could also be due to degradation in the matrix. With a smaller volume of aqueous sample required compared to solid phase extraction (SPE), SPME is an attractive method for the analysis of limited volumes of sediment pore-water. Moreover, the use of SPME eliminated interferences present in sediment extracts thus allowing the detection of the target analytes that were otherwise difficult to detect by direct injection. PMID- 15794570 TI - Development and characterization of a solvent extraction-gas chromatographic/mass spectrometric method for the analysis of perfluorooctanesulfonamide compounds in solid matrices. AB - A method utilizing solvent extraction and analysis by gas chromatography-positive chemical ionization mass spectrometry (SE-GC-PCIMS) was developed for the analysis of three neutral hydrophobic perfluorooctanesulfonamide compounds [perfluorooctanesulfonamide (PFOSA), N-ethyl perfluorooctanesulfonamide (N EtPFOSA), and N,N-diethyl perfluorooctanesulfonamide (N,N-Et2PFOSA)]. These compounds are suspected metabolic precursors of perfluorooctane sulfonate. The SE GC-PCI-MS method was used to analyze all three perfluorooctanesulfonamides in fast food, fish, and Arctic marine mammal liver samples. The SE-GC-PCI-MS method produced relatively higher recoveries of the analytes (averaging 83 +/- 6%, 84 +/ 9%, and 89 +/- 19% for N,N-Et2PFOSA, N-EtPFOSA, and PFOSA, respectively) with lower coefficients of variation, and less susceptibility to matrix effects, than ion pair extraction-liquid chromatography-tandem mass spectrometric methods. Method detection limits (MDLs) were 100, 120, and 250 pg/g for N,N-Et2PFOSA, N EtPFOSA, and PFOSA, respectively. The three compounds were found at concentrations ranging from below the MDL to 22 ng/g wet weight in fast food, fish, and Arctic marine mammal liver samples. PMID- 15794572 TI - Organic solvent high-field amplified stacking for basic compounds in capillary electrophoresis. AB - Many water-miscible organic solvents, especially acetonitrile and acetone, bring along significant degrees (approximately 30 times) of stacking by electroinjection through high-field amplified injection for the basic compounds compared to that for aqueous buffers or water. The relative stacking of different compounds in acetonitrile or acetone is different compared to that for water. Stacking by electroinjection in organic solvents is less stringent and easier to accomplish in practice. Acids and salts, in aqueous solutions, can ruin the stacking for both organic and aqueous solvents; however, this effect can be better tolerated by diluting the sample in acetonitrile. Thus, this stacking is termed "organic solvent high-field amplified injection". This stacking by electroinjection is enhanced by increasing the electrophoresis buffer concentration and can be better than that by pressure injection. From the practical aspects, some cationic drugs present in serum such as amiodarone can be detected at the therapeutic levels by electroinjection on the capillary after protein precipitation by acetonitrile. PMID- 15794571 TI - Capillary electrophoresis immunoassay based on an on-column immunological reaction. AB - An on-column immunological reaction was employed to achieve simple and rapid analysis in an immunoassay based on capillary electrophoresis using semiconductor laser-induced fluorescence detection. Human serum albumin (HSA) labeled with sulfoindocyanine succinimidyl ester (Cy5), a fluorescent compound with an absorption maximum at 649 nm, was used as a fluorescent probe for the immunoassay. In a binding assay, with anti-HSA as the analyte molecule, Cy5-HSA was injected in a capillary column followed by the injection of anti-HSA so as to form individual zones. By applying a potential, the anti-HSA reacted with Cy5-HSA at the boundary between Cy5-HSA and anti-HSA zones, since anti-HSA has a higher electrophoretic mobility than Cy5-HSA. Furthermore, the on-column method enhances the sensitivity by injecting a large volume of the sample. Free Cy5-HSA and its immunocomplex with anti-HSA were separated with less degradation in resolution than that predicted from the injection time of anti-HSA, even when the injection time for anti-HSA was increased. The ratio of the peak area of the complex to that of the total Cy5-HSA (free Cy5-HSA and the complex) increased in proportion to the injection time of anti-HSA. As a result, the detection limit was improved up to eight-fold (the concentration detection limit, 0.007 mg mL(-1), for an injection time of 240 s, compared to that obtained using an off-column sample preparation. Furthermore, the on-column reaction method was applicable to an immunoassay to determine native HSA, in which native HSA and Cy5-HSA react with anti-HSA stepwise. The detection limit in the stepwise reaction immunoassay was 0.005 mg mL(-1), which is 14 times lower than that in an off-column method, with the analysis time less than 10 min as the result of increasing the injection time of native HSA. In addition, the present on-column immunoassay was applied to the sample containing a high concentration of salts for investigating the effect of salts in the sample solution. PMID- 15794573 TI - On-line cation-exchange preconcentration and capillary electrophoresis coupled by tee joint interface. AB - An on-line preconcentration method based on ion exchange solid phase extraction was developed for the determination of cationic analytes in capillary electrophoresis (CE). The preconcentration-separation system consisted of a preconcentration capillary bonded with carboxyl cation-exchange stationary phase, a separation capillary for zone electrophoresis and a tee joint interface of the capillaries. Two capillaries were connected closely inside a 0.3 mm i.d. polytetrafluoroethylene tube with a side opening and fixed together by the interface. The preparations of the preconcentration capillaries and interface were described in detail in this paper. The on-line preconcentration and separation procedure of the analysis system included washing and conditioning the capillaries, loading analytes, filling with buffer solution, eluting analytes and separating by capillary zone electrophoresis (CZE). Several analysis parameters, including sample loading flow rate and time, eluting solution and volume, inner diameter and length of preconcentration capillary etc., were investigated. The proposed method enhanced the detection sensitivity of CE-UV about 5000 times for propranolol and metoprolol compared with normally electrokinetic injection. The detection limits of propranolol and metoprolol were 0.02 and 0.1 microg/L with the proposed method respectively, whereas those were 0.1 and 0.5 mg/L with conventional electrokinetic injection. The experiment results demonstrate that the proposed technique can increase the preconcentration factor evidently. PMID- 15794574 TI - Rapid evaluation of nickel binding properties of His-tagged lactate dehydrogenases using surface plasmon resonance. AB - The use of surface plasmon resonance (SPR), for the comparison of metal binding properties of polyhistidine tags, was evaluated. Six different tags containing various number of histidines, either none (tags n and t), three (tags H3A3 and HA2HA2H) or six (tags H6 and His6), were genetically fused to the N-terminal of lactate dehydrogenase (LDH). The binding ability of these constructs to nickel ions, immobilised with nitrilotriacetic acid (NTA), was tested both by conventional immobilised metal ion affinity chromatography (IMAC) and SPR. The relative binding strengths of the tags to nickel were identical using both methods (n approximately t < HA2HA2H < H3A3 < His6 < H6), confirming the value of the SPR technique for investigating metal-protein interactions. Protein modelling has also proved to be useful in supporting the experimental results. PMID- 15794575 TI - Vapor-phase analysis of isobutyl acetate, isopropyl acetate, n-propyl acetate and their respective alcohols using solid-phase microextraction-gas chromatography with a mass selective detector. AB - A solid-phase microextraction (SPME)-GC-MS method for three esters and the corresponding alcohols was tested for responses in accuracy, within-run precision (repeatability), and between-run precision (reproducibility) due to individual operators, individual analysis days, and differing analyte concentrations. At 5 ppm (v/v) [ppmv], three of the six analytes showed significant (p < 0.05) operator effects, while five of six analytes gave a significant effect due to the days of analysis. At 20 ppmv, five of the six analytes gave significant operator and daily effects. At 100 ppmv, all the analytes showed significant daily effects but no operator effects were observed. The repeatability was concentration dependent, with all six analytes combining for an average RSD of 12.1 +/- 6.1% at 1 ppmv, becoming most precise at 50 ppmv at 1.01 +/- 0.45%, then increasing at 100 ppmv to 4.12 +/- 1.88%. The contributors to error trended as: concentration > daily effects > operator. PMID- 15794576 TI - Metal tubing/frit with a sintered frit of silica particles and a chromatography column with such tubing/frits. AB - This study is about the metal tubing/frit prepared by treating a powdered silica with a strong base, drying it, scrubbing it into powders, putting it in the tip of metal tubing at a proper depth, and sintering the powders to form a durable sintered frit. This study is also about a liquid chromatographic column prepared by installing the metal tubing/frit at the column inlet and outlet unions and by placing the column main body tubing packed with a stationary phase between the two unions. The metal tubing/frit made according to the description is easily replaced and fixed without disassembling the column when it is necessary. The column of this design could be a cheap substitute for the conventional microcolumn. PMID- 15794577 TI - Preparative separation and purification of deoxyschisandrin and gamma-schisandrin from Schisandra chinensis (Turcz.) Baill by high-speed counter-current chromatography. AB - High-speed counter-current chromatography (HSCCC) was successfully applied to the preparative separation and purification of deoxyschisandrin and gamma-schisandrin from the crude petroleum ether extracts of Schisandra chinensis (Turcz.) Baill. The optimum solvent system composed of n-hexane-methanol-water (35:30:3, v/v) led to the successful preparation of deoxyschisandrin and gamma-schisandrin. The analysis of HPLC for each peak fraction of preparative HSCCC showed that the purity of deoxyschisandrin (8 mg) was over 98% and gamma-schisandrin (12 mg) was over 96% from 100 mg of the crude petroleum ether extracts in one-step separation. PMID- 15794578 TI - Separation and purification of baicalin and wogonoside from the Chinese medicinal plant Scutellaria baicalensis Georgi by high-speed counter-current chromatography. AB - A preparative high-speed counter-current chromatography (HSCCC) method for isolation and purification of baicalin and wogonoside from the Chinese medicinal plant Scutellaria baicalensis Georgi (Huang-qin in Chinese) was successfully established by using ethyl acetate-methanol-1% acetic acid water (5:0.5:5, v/v) as the two-phase solvent system. The upper phase of ethyl acetate-methanol-1% acetic acid water (5:0.5:5, v/v) was used as the stationary phase of HSCCC. Baicalin (58.1 mg) and wogonoside (17.0mg) with the purity of 99.2 and 99.0%, respectively, were separated successfully in one-step separation from 120 mg of crude sample from S. baicalensi, Georgi. The structures of baicalin and wogonoside were identified by 1H NMR and 13C NMR. PMID- 15794579 TI - Gas chromatographic method for the analysis of allelopathic natural products in rye (Secale cereale L.). AB - Accurate and reproducible methods for the analysis of plant allelochemicals are a requirement for the study of chemical interactions between plants. This paper describes a method for sample preparation and quantitative analysis of the allelopathic chemical content of rye (Secale cereale L.) using gas chromatography (GC). Sample preparation consists of extraction of freeze-dried rye vegetative tissue with aqueous ethanol followed by partitioning of the allelochemicals into ethyl acetate, evaporation, and derivatization using the trimethylsilylating reagent N-methyl-N-trimethylsilyltrifluoroacetamide. GC analysis of the silylated mixture was performed using flame ionization detection. This method permits analysis of all known rye allelopathic agents including 2,4-dihydroxy-1,4 benzoxazin-3-one, its corresponding glucoside, 2-benzoxazolinone, beta hydroxybutyric acid, and beta-phenyllactic acid. Identities of all compounds were confirmed by GC/MS analysis. PMID- 15794580 TI - Capillary zone electrophoresis separation of enantiomers of lisuride. AB - Lisuride is an ergot alkaloid derivative with dopaminergic activity. It is used for treatment of Parkinsonism and some other diseases associated with high level of prolactine. Lisuride is a chiral compound derived from natural ergot alkaloids. A new capillary zone electrophoresis (CZE) method capable of separating the enantiomers of lisuride was developed. Using the optimized conditions (acidic electrolyte with the addition of gamma-cyclodextrin (gamma CD)) as low as 0.02% of undesirable L-lisuride can be detected. Selected method characteristics, i.e., linearity (0-20 mg/l), precision (2.0% at 5 mg/I), and accuracy (101 +/- 4% at 5 mg/l) were evaluated. The optimized method was applied for the analysis of real batches of Lisuride hydrogenmaleate and Lisuride base manufactured by IVAX Pharmaceuticals. It was found that they contain less than 0.02% of undesirable L-enantiomer. PMID- 15794581 TI - [The Revista Espanola de Salud Publica becomes friend of the woods]. PMID- 15794582 TI - [A method for ascertaining the seasonal pattern of hospital emergency department visits]. AB - A suitable method is set forth in this study for isolating and describing high frequency seasonal variations which may occur in some sets of time series in the healthcare setting. If the simultaneous presence of seasonal variations within a day, a week and a year is not taken into account, the estimates of seasonal behaviours may be noticeably distorted and the decisions made by those in charge of managing healthcare services based thereupon might be erroneous. The proposed procedure, based upon taking simultaneous periodic changes jointly into consideration is applied to an hourly series of patients visiting an hospital emergency room in Tenerife. The results obtained indicate that the most frequent visits take place in February and March, Mondays being the days on which the greatest demand occurs and the early morning hours of the day showing the minimum demands. A long-term growth is also observed. And, above all, the proposed method is shown to provide some advantages over other approaches in which each one of the seasonal variations is estimated as if the others do not exist. PMID- 15794583 TI - [Characteristics of 107 Spanish healthcare registries and evaluation of the utilization thereof]. AB - BACKGROUND: The information concerning the currently existing healthcare registries in our country is not readily accessible, is scarce and some are not well-known, possibly being underused in comparison to their many potential uses. This study is aimed at evaluating the characteristics of the Spanish Healthcare registries and the utilization thereof, especially in Healthcare technology assessment. METHODS: Descriptive, cross-sectional study of the Spanish Healthcare registries within the 1997-2002 period. These registries were identified by means ofa bibliographic databases search and by way of a survey of Scientific Societies and Central and Autonomic Healthcare Administration. Another bibliographic databases search was conducted of the publications which use data from the healthcare registries included and have been evaluated by applying the scientific evidence quality criteria. RESULTS: A total of 107 healthcare registries were identified, most of which of local or regional coverage (71%), preferably were under Autonomic government authority (64.5%). The areas showing the largest number of registries identified were those related to death statistics (16.8%) and cancer (15.9%). A total of 298 publications were retrieved which analyze data produced by the registries identified, most of which are devoted to the study of the frequency and distribution of the events recorded (58.1%) and less frequently to the conducting of healthcare technology assessment studies (24.4%). CONCLUSIONS: The critical evaluation of the publications made it possible to identify some elements related to the analysis methodology and design which would heighten the quality of the healthcare technology assessment. Healthcare registries in Spain have developed recently and to differing degrees. Important areas without any records and improvement elements related to the use of healthcare registries for healthcare technology assessment were detected. It would be advisable to avail of a register of registries which would provide relevant, up-dated information thereon. PMID- 15794584 TI - [Perceived health and use of health care services in inmates of an Andalusian prison, 1999]. AB - BACKGROUND: The study of the health condition of the populations under confinement in penitentiaries is based on the evidence of a more deteriorated health than the general population and a greater degree of social exclusion, which is associated with worse general health. This study is aimed at ascertaining how the inmates of an Andalusian penitentiary perceive their health condition and the use made thereby of the healthcare services, as well as the factors associated with those variables. METHODS: Descriptive, cross-sectional study. The data was collected with a questionnaire. The sample size was 450 inmates, 90.4% of whom were males. RESULTS: Seventy-two percent of those taking part in the study considered their health to be good or very good, 32.7% stating having seen the doctor once a month or more often. A total 43.1% of the participants stated having chronic illnesses, mainly HIV (19.1%) and hepatitis C (18.2%); 40.9% stating that they take medication. Worse health was perceived among the older inmates, those who have to serve longer than a five-year sentence, those who are repeat offenders and those not having pending trials. Those perceiving their health to have deteriorated over the past year have chronic illnesses and take medication. The regression model for the use of healthcare services shows that they are used to a greater extent by those who are serving a longer than a 5-year sentence, those who have been in prison for less than a year and those who take medication. CONCLUSIONS: The results show the importance of increasing the monitoring of possible addictive disorders upon entering prison and of the trend and treatment of chronic diseases. PMID- 15794585 TI - [Outbreak of Salmonella enteriditis food poisoning at a dining hall facility]. AB - BACKGROUND: An outbreak of Salmonella enteriditis food poisoning having occurred at two banquet halls, affecting eight groups of guests, is described. This research was aimed at determining the food by means of which this illness was transmitted. METHODS: An analysis was conducted of the food processing procedures subject to having caused the outbreak. The association of the foods with the illness was analyzed by means of a case and control design. The adjusted Odds Ratio (aOR) and the 95% confidence intervals (C195%) were calculated by logic regression. RESULTS: The guests totalled 1771 in number, being divided up into 13 groups for three days running. A total of 629 individuals (36%) were surveyed The probable cases totalled 250 in number, 61 confirmed by Salmonella enterica. The glazed biscuit (dessert with raw egg having undergone no heat processing) showed the highest adjusted Odds Ratio (aOR = 20.40; CI95%:7.52-55.30) and tested positive for Salmonella enterica of the enteritidis serotype. CONCLUSIONS: Epidemiological and laboratory evidence supports the glazed biscuit having been the contaminated food having caused the outbreak. The research stressed as contributing factors: the use of raw egg, the production of large quantities ahead of time, prior to the time at which the food item in question was to be eaten. PMID- 15794586 TI - [Vascular by-pass surgery infection incidence and risk factors at the Juan Canalejo Hospital in Corunna within the 2000-2002 period]. AB - BACKGROUND: Vascular by-pass surgery is being used increasingly more frequently on elderly patients entailing several risk factors. Surgical infections in these procedures are a devastating complication which is related to high morbility and death rates. This study is aimed at ascertaining the incidence of surgical infections among patients having undergone a by-pass procedure with vascular prostheses, the risk factors associated thereto and the type or prosthesis entailing a lower infection rate. METHODS: A retrospective cohort study was conducted at the Vascular Surgery Unit of the Juan Canalejo University Hospital throughout the 2000-2002 period. The Disease Control Center infection criteria were those applied. The frequency measurements calculated were cumulative incidence and relative risk as an association measurement. The statistical tests employed were the Chi-square and the Chi-square trend tests, the Student's-T for univariate analysis and logic regression for the multi-variate analysis. RESULTS: The cumulative incidence rate for the 2000-2002 period was 22.6% to 12.1%. The most frequent microorganisms were Staphylococcus aureus, meticilin-resistant Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli. The risk factors related to infection found were female sex (RR=1.8) anesthetic risk >2 (RR=1.7) and patient implanted with sapheneous vein vascular prosthesis (RR=3.8). CONCLUSIONS: The risk factors identified were the female sex, anesthetic risk and the type of prosthesis used, all of which condition infection-related complication in by-pass surgery. PMID- 15794587 TI - [Differences in the use of family planning methods by adolescent females according to the education model utilized during pregnancy. Monterrey, Mexico]. AB - BACKGROUND: The objective was to compare the use of family planning methods during the immediate postpartum period and two years following childbirth among the adolescent females who had followed two different intervention programs during their pregnancies. METHODS: A quasi-experimental study was designed. A total of 62 pregnant adolescents were selected to comprised two intervention groups, the PRECEDE model and the Health Belief model (MCS) groups. Non parametric statistical tests were employed and 95% confidence intervals estimated. RESULTS: The average starting knowledge in the MCS groups was 69.12 points (95% CI 63.27-74.97) and ending 89.71 points (95% CI 86.24-93.17), while the starting knowledge for the PRECEDE group was 49.39 points (95% Cl 42.24 56.54) and ending 75.25 points (95% CI 71.12-79.38). IN the immediate postpartum, 93% (95% CI 83.5-100) of the adolescents in the PRECED group accepted the use of a family planning method similar to that employed by the MCS group, of 94.2 (95% CI 86.3-100). As regards the continued use of the method, that is, two years later, the PRECEDE strategy had a greater effect that the MCS strategy, respectively 92% (95% Cl 82-100) and 72% (95% CI 56.9-87.1). CONCLUSIONS: A difference was found to exist between models as regards the use of family planning methods 2 years following childbirth. The PRECEDE program is proposed as the education strategy for preventing a second pregnancy among adolescent females. PMID- 15794588 TI - Effect of endocrine disrupters on macrophage functions in vitro. AB - In previous papers we reported that endocrine disrupters (EDs) such as diethyl stilbesterol (DES), 4,4'-isopropylidene diphenol (Bisphenol A: BPA), bis (2 ethylhexyl) phthalate (EHP) and p-nonylphenol (NP) stimulated proliferative responses and cytokine productions of murine spleen cells and thymus cells in vitro. In this paper we report the effect of EDs on murine macrophage functions in vitro. DES, BPA, EHP and NP stimulated thioglycolate-induced peritoneal exudate cells (macrophages) to produce cytokines such as interleukin (IL)-1, IL 6, IL-12, tumor necrosis factor and macrophage chemotactic protein- 1. The effect of these EDs was observed at the concentrations of 10(-6)-10(-8) M, and the maximum response was obtained at 10(-7 M.) The accessory function for T cell activation and CD86 expression, an important molecule for accessory function, was enhanced in EDs-treated macrophages. These results suggest that EDs have modulating activities of macrophage functions. PMID- 15794589 TI - Effect of diesel exhaust on immune responses in C57BL/6 mice intranasally immunized with pollen antigen. AB - To investigate the effect of diesel exhaust or particle-free diesel gas on immune responses in IgE low responder mice, C57BL/6 mice immunized intranasally with sugi basic protein were exposed to diesel exhaust or diesel gas components. We evaluated the changes in lymphocyte subpopulations, cell proliferation, chemokine production of cervical lymph nodes cells, and antigen-specific-antibody levels in plasma. Exposure to diesel gas decreased the percentage of CD4+ and TCR-beta+ T cells of cervical lymph nodes from immunized mice. Culture supernatants of cervical lymph nodes cells from diesel gas-exposed mice had significantly increased levels of macrophage inflammatory protein-1alpha and thymus and activation-regulated chemokine, but exposure to diesel exhaust did not affect it. Antigen-specific IgG2a titers in plasma were significantly enhanced after their exposure to diesel exhaust or gas. In contrast, exposure to diesel exhaust or gas markedly decreased antigen-specific IgG1 titers in immunized mice. These facts indicate that concurrent exposure to allergen and diesel exhaust or diesel gas modulates chemokine production in cervical lymph nodes cells and antibody production in plasma differentially. PMID- 15794591 TI - [An investigation of social support networks for working mothers]. AB - The purpose of our study was to investigate the social support network for working mothers, including physical, emotional and informational sources, and to discuss strategies to reduce stress caused by their multiple role strain. We asked 18 participants to fill in a questionnaire and conducted semi-constructed interviews. The results of this study showed that more than half of the working mothers want to continue their career because of self growth and the rewards from their job. Many mothers responded that a sympathetic understanding by their husbands, families and people at workplaces is essential to create a balance between their career and housework. As a result, their husbands mainly provide physical support in housekeeping and child caring, whereas their mothers provide physical and emotional support from knowledge based upon their own experience. Almost all working mothers complained of a shortage of nurseries and child caring facilities as well as limited availability of flexible services. In conclusion, the results of our study suggested that the identity of the traditional gender role needs to be changed to a contemporary one, like a husband fully participating in his child's rearing activities by reconsidering his working hours. Moreover, community services to support working mothers should be increased. PMID- 15794590 TI - A very low level of magnetic field exposure does not affect a participant's mental fatigue and stress as much as VDT work. AB - Exposure to Magnetic Fields (MF) may affect the health of workers in an occupational environment. However, there is no clear evidence that the emission of MF from the visual display terminals (VDT) affects workers'mental status or induces stress in office environments. The purpose of our study was to investigate whether exposure to the MF during VDT work affects mental fatigue or induces stress related to psycho-physiological changes in workers. Thirty-seven students were voluntarily recruited to participate in this study. We created experimental conditions that a pair of the liquid crystal display (LCD) terminals separated by screens was closely placed in front of the visual suface of the cathode ray tube (CRT) units. All participants were not able to recognize whether the power of the CRT Units was ON or OFF while they worked with the LCD terminals. When the CRT unit was ON, participants who worked with the LCD terminal were called CRT-ON group. On the other hand, when the CRT unit was OFF, participants who worked with the CLD terminal were called CRT-OFF group. Incredibly short profiles of mood states (ISP)and subjective symptom questionnaires were used as indices of the mental fatigue as well as psychological stress before and after VDT work. Furthermore, salivary chromogranin A and urinary 8-hydroxydeoxyguanosine (8-OH-dG) were collected to measure physiological changes before and after VDT work. The results of the questionnaires and salivary and urinary stress markers showed that the exposure to the MF during VDT work did not significantly induce mental fatigue or psychological stress in comparison with the CRT-OFF group. Contrary to the results from the exposure, mood states of vigor decreased greatly (P < 0.05) whereas the states of fatigue and confusion increased significantly (P < 0.001) after VDT work. In the questionnaires for subjective symptoms, participants complained remarkably regarding 14 out of 25 conditions after VDT work. Moreover, urinary 8-OH-dG excretion increased significantly after VDT work. Taken together, the results of this study did not detect any bad health effects of MF exposure on the VDT participants but suggested that VDT work itself affects their mental fatigue and psycho-physiological status. PMID- 15794593 TI - [Relationship between bone metabolism and effects of lifestyles after gastrectomy]. AB - The aim of this study was to examine the effects of bone metabolism impairment after a gastrectomy, as well as the effects of dietary life and physical activity on bone density, in order to obtain basic information on lifestyles that prevent osteoporosis. The study subjects were 27 male patients who had been diagnosed as having gastric cancer at a university hospital and had received a gastrectomy. All subjects gave their informed consent. The level of the bone metabolism impairment was examined by bone cell density measurement, ultrasound densitometer and by the measurements of bone metabolism parameters, i.e., BAP, DPD, NTx, corrected Ca. Physical activity was measured by the number of steps, and dietary life was investigated using a questionnaire. The SOS value, which is an index to the bone metabolism impairment, decreased 3 weeks after the gastrectomy. Although the SOS value recovered once, it decreased after 6 months again. Afterwards, it recovered to the preoperative level in 12 months. The levels of BAP, which are the osteogenesis markers, significantly decreased 3 weeks after the gastrectomy and increased three months later. DPD level, which is a bone resorption marker, increased significantly. The effect of physical activity and dietary life on the bone density was not recognized. Though the bone metabolism impairment appeared 3 weeks after the gastrectomy, the bone formation process after bone resorption was affected by individual differences. Consequently, it was impossible to clarify the factors of lifestyles which affect bone metabolism impairment. PMID- 15794592 TI - [Body cooling capacity of water circulated cooling pants during lower body exercise in a hot environment]. AB - We investigated the body cooling capacity of water-circulated cooling pants during lower body exercise in a hot environment. Five subjects were asked to ride a bicycle ergometer and to keep a work rate of 75 W continuously for 20 minutes. They then rest on the ergometer for 15 minutes, in a climate chamber kept at WBGT = 31.5 degrees C. They repeated the same procedure three times under three different conditions; 1) wearing cooling pants with cold water supply; 2) wearing cooling pants without water supply; 3) wearing short pants instead of cooling pants. Esophageal temperature and heart rate of the subjects, when their cooling pants were supplied with cold water, were lower compared with the condition without water supply, and were almost the same as the condition wearing short pants. Water-circulated cooling pants should effectively cool the body at work where the worker has to wear pants that cover his legs to prevent injury. PMID- 15794594 TI - [Two cases of nonrecurrent inferior laryngeal nerve--different branching levels from vagus nerve]. AB - We observed two cases of nonrecurrent inferior laryngeal nerve (NRILN). Case 1, a 71 year old man was diagnosed as having papillary carcinoma. NRILN was found during his operation. It directly branched from the right cervical trunk of the vagus nerve at the level of the cricoid cartilage and then entered the larynx after running behind the thyroid gland. Case 2, a 64 year old woman was diagnosed as having primary hyperparathyroidism. In this patient, the NRILN branched at the level of the inferior pole of the thyroid gland, rose up beside the tracheal wall and entered the larynx. In both patients, preoperative CT scan and postoperative MR angiography revealed the aberrant right subclavian artery. A postoperative barium swallow test showed the compression of the esophagus by this anomalous artery in case 1. Although it is possible to predict the presence of NRILN by preoperative imaging tests, the branching level from the vagus nerve is unpredictable. Surgery must be performed with this point in mind, if the presence of NRILN is suspected. PMID- 15794595 TI - [Personal exposures of benzene treated workers and a simple biological monitoring]. AB - A simple method of biological monitoring has been developed for occupational benzene exposure. Personal benzene exposure monitoring using a passive sampler and GC/FID was carried out on 74 workers from a benzene-treated company. Their urines were collected before and after work-shift. After treatment of urine samples using solid phase extraction (SPE), trans, trans-muconic acid(t, t-MA) concentration in the elute was analysed by HPLC. Correlation between benzene exposure (X: ppm) and urinary t, t-MA concentration (Y: mg/g x creatinine) for non-smokers was Y = 0.948X + 0.586 (r = 0.798, P < 0.01) and Y = 0.885X + 0.894 (r = 0.871, P < 0.01) for smokers, respectively. The t, t-MA concentration on 1 ppm TLV exposure to benzene was estimated as 1.5 and 1.8 (mg/g creatinine) for non-smokers and smokers, respectively. These values are in agreement with some investigators. This indicates that our simple method for biological monitoring of benzene exposure can be of great service. PMID- 15794596 TI - Vibration-assisted electron tunneling in C140 transistors. AB - We measure electron tunneling in transistors made from C(140), a molecule with a mass-spring-mass geometry chosen as a model system to study electron-vibration coupling. We observe vibration-assisted tunneling at an energy corresponding to the stretching mode of C(140). Molecular modeling provides explanations for why this mode couples more strongly to electron tunneling than to the other internal modes of the molecule. We make comparisons between the observed tunneling rates and those expected from the Franck-Condon model. PMID- 15794597 TI - Spin-polarized light-emitting diodes with Mn-doped InAs quantum dot nanomagnets as a spin aligner. AB - We have fabricated and characterized surface-emitting, spin-polarized light emitting diodes with a Mn-doped InAs dilute magnetic quantum dot spin-injector and contact region grown by low-temperature molecular beam epitaxy, and an In(0.4)Ga(0.6)As quantum dot active region. Energy-dispersive X-ray and electron energy loss spectroscopies performed on individual dots indicate that the Mn atoms incorporate within the dots themselves. Circularly polarized light is observed up to 160 K with a maximum degree of circular polarization of 5.8% measured at 28 K, indicating high-temperature spin injection and device operation. PMID- 15794599 TI - Energy conversion efficiency in nanotube optoelectronics. AB - We present theoretical performance estimates for nanotube optoelectronic devices under bias. Current-voltage characteristics of illuminated nanotube p-n junctions are calculated using a self-consistent nonequilibrium Green's function approach. Energy conversion rates reaching tens of percent are predicted for incident photon energies near the band gap energy. In addition, the energy conversion rate increases as the diameter of the nanotube is reduced, even though the quantum efficiency shows little dependence on nanotube radius. These results indicate that the quantum efficiency is not a limiting factor for use of nanotubes in optoelectronics. PMID- 15794598 TI - Exciton localization in vertically and laterally coupled GaN/AlN quantum dots. AB - Near-field and time-resolved photoluminescence measurements show evidence of exciton localization in vertically and laterally coupled GaN quantum dots (QDs). The binding energies in multiple period QDs (MQDs) are observed to be stronger by more than six times compared to single period QDs (SQDs). Excitons in MQDs have a short (450 ps) lifetime and persist at room temperature, while SQDs exhibit extraordinarily long (>5 ns) lifetime at 10 K due to reduced spatial overlap of electron and hole wave functions in strained QDs. PMID- 15794600 TI - Imaging a single-electron quantum dot. AB - Images of a single-electron quantum dot were obtained in the Coulomb blockade regime at liquid He temperatures using a cooled scanning probe microscope (SPM). The charged SPM tip shifts the lowest energy level in the dot and creates a ring in the image corresponding to a peak in the Coulomb-blockade conductance. Fits to the line shape of the ring determine the tip-induced shift of the energy of the electron state in the dot. SPM manipulation of electrons in quantum dots promises to be useful in understanding, building, and manipulating circuits for quantum information processing. PMID- 15794601 TI - Structures and stability of water nanoclusters in hydrophobic nanospaces. AB - The temperature dependencies of in situ small-angle X-ray scattering (SAXS) of water adsorbed and of adsorption isotherm of water in hydrophobic carbon nanopores were measured over the temperature range of 293 to 328 K. The structures of water nanoclusters adsorbed in the nanopores were determined with the density fluctuation analysis of in situ SAXS data. The difference of the density fluctuations between adsorption and desorption was ascribed to the water structural difference. The structural transitions of the water nanoclusters were observed around 318 K for adsorption and 308 K for desorption. PMID- 15794602 TI - Template-synthesized protein nanotubes. AB - A layer-by-layer deposition strategy for preparing protein nanotubes within the pores of a nanopore alumina template membrane is described. This method entails alternately exposing the template membrane to a solution of the desired protein and then to a solution of glutaraldehyde, which acts as cross-linking agent to hold the protein layers together. The number of layers of protein that make up the nanotube walls can be controlled at will by varying the number of alternate protein/glutaraldehyde cycles. After the desired number of layers have been deposited on the pore walls, the alumina template can be dissolved to liberate the protein nanotubes. We show here that glucose oxidase nanotubes prepared in this way catalyze glucose oxidation and that hemoglobin nanotubes retain their heme electroactivity. Furthermore, for the glucose oxidase nanotubes, the enzymatic activity increases with the nanotube wall thickness. PMID- 15794603 TI - Electrical docking of microtubules for kinesin-driven motility in nanostructures. AB - We demonstrate localized electrical control of the docking of microtubules onto engineered kinesin-coated structures. After applying a voltage to a gold electrode, we observe an enhanced transport of microtubules from solution toward the surface and a subsequent increase of the amount of moving microtubule shuttles. Switching off the voltage leads to a partial detachment of microtubules from the surface. The surface coverage of microtubules, during both the docking and undocking events, follows an exponential time dependence. We provide a simple kinetic model, incorporating the equilibrium between free and surface-bound microtubules, that explains these data. PMID- 15794604 TI - Biological assembly of nanocircuit prototypes from protein-modified CdTe nanowires. AB - CdTe nanowires made by self-organization of CdTe nanoparticles in aqueous media were separately conjugated with complementary biological connectors, such as antigen-antibody and biotin-streptavidin. Transmission electron microscopy images and Forster resonance energy transfer measurements in nanowire superstructures with different diameters indicate that biological affinity of the attached proteins results in the formation of crossbar and end-to-side connections between the nanowires. A prototype of a logical circuit made from a triangular arrangement of the nanowires spontaneously assembled on a Si substrate was examined by conducting atomic force microscopy. While diode-like behavior was observed in the sides of the triangle, the nanowire junction points were found to be nonconductive. It was attributed to high tunneling barrier created by protein molecules wedged between the nanowires. Suggestions are made how to reduce it or use the insulating gap between the nanowires as a framework for single-electron devices. PMID- 15794606 TI - Imaging surface plasmon of gold nanoparticle arrays by far-field Raman scattering. AB - We present Raman spectra and Raman images of the methylene blue molecule adsorbed as a single layer on gold nanoparticles regularly arranged in periodic arrays. Spectra and images are recorded in the same spatial and spectral regions using an excitation under total internal reflection. Images of the Raman scattering appear as spots of circular shape located at the particle positions with size defined by the diffraction limit. It appears that all excited particles contribute equally to the Raman signal if the Gaussian intensity distribution of the laser beam is taken into account. These results demonstrate that Raman scattering can be a useful technique to study plasmon properties. PMID- 15794605 TI - Presentation and recognition of biotin on nanofibers formed by branched peptide amphiphiles. AB - A branched peptide amphiphile system was designed for enhanced recognition of biotin on nanofibers formed by self-assembly of these molecules. Branching at a lysine residue was used to design peptide amphiphiles that are capable of presenting more than one epitope per molecule. We found that biotinylated branched structures form nanofibers that enhance recognition by the avidin protein receptor relative to similar nanostructures formed by linear peptide analogues. Biotin-avidin binding to the supramolecular nanofibers was characterized by measurement of fluorescence from nanofibers incubated with chromophore-conjugated avidin. PMID- 15794607 TI - Assembly of silica nanowires on silica aerogels for microphotonic devices. AB - We report on the assembly of low-loss silica nanowires into functional microphotonics devices on a low-index nondissipative silica aerogel substrate. Using this all-silica technique, we fabricated linear waveguides, waveguide bends, and branch couplers. The devices are significantly smaller than existing comparable devices and have low optical loss, indicating that the all-silica technique presented here has great potential for future applications in optical communication, optical sensing, and high-density optical integration. PMID- 15794608 TI - Deflection of nanotubes in response to external atomic collisions. AB - The mechanical response of single-walled and multiwalled carbon nanotubes to a series of external Ar atom impacts is examined with classical molecular dynamics simulations. The extent to which the carbon nanotubes deform in the direction perpendicular to their axis is found to depend on the amount of momentum transferred during the collisions. The details of the mechanical response and recovery of the nanotubes after release are also found to depend on the nanotube configurations. PMID- 15794609 TI - Kelvin probe force microscopy study on conjugated polymer/fullerene bulk heterojunction organic solar cells. AB - We conducted a comprehensive Kelvin probe force microscopy (KPFM) study on a classical organic solar cell system consisting of MDMO-PPV/PCBM blends. The KPFM method yields the information of topography and local work function at the nanometer scale. Experiments were performed either in the dark or under cw laser illumination at 442 nm. We identified distinct differences in the energetics on the surface of chlorobenzene and toluene cast blend films. Together with high resolution scanning electron microscopy (SEM) experiments we were able to interpret the KPFM results and to draw some conclusions for the electron transport toward the cathode in the solar cell configuration. The results suggest that surfaces of toluene cast films exhibit a morphologically controlled hindrance for electron propagation toward the cathode, which is usually evaporated on top of the films in the solar cell device configuration. PMID- 15794610 TI - Controlled fabrication of silicon nanowires by electron beam lithography and electrochemical size reduction. AB - We demonstrate that electrochemical size reduction can be used for precisely controlled fabrication of silicon nanowires of widths approaching the 10 nm regime. The scheme can, in principle, be applied to wires defined by optical lithography but is here demonstrated for wires of approximately 100-200 nm width, defined by electron beam lithography. As for electrochemical etching of bulk silicon, the etching can be tuned both to the pore formation regime as well as to electropolishing. By in-situ optical and electrical characterization, the process can be halted at a certain nanowire width. Further electrical characterization shows a conductance decreasing faster than dimensional scaling would predict. As an explanation, we propose that charged surface states play a more pronounced role as the nanowire cross-sectional dimensions decrease. PMID- 15794611 TI - The formation of supported lipid bilayers on silica nanoparticles revealed by cryoelectron microscopy. AB - The controlled fabrication of biocompatible devices made of lipid bilayers deposited onto flat solid supports presents interest as models of cell membranes as well as for their biotechnological applications. We report here on the formation of supported lipid bilayers on silica nanoparticles (nanoSLBs). The successive steps of the adsorption of lipid vesicles on nanoparticles and the formation of nanoSLBs are revealed in detail by cryotransmission electron microscopy (cryo-EM). The formation of nanoSLBs was achieved for liposomes with positive, neutral, and low net negative charge, while liposomes with a high net negative charge adsorbed to silica nanoparticles but did not rupture. The nanoSLBs were found to follow faithfully the surface contours of the particles, information yet unavailable for SLB formation on planar solid substrates. PMID- 15794612 TI - Coulomb oscillations and Hall effect in quasi-2D graphite quantum dots. AB - We perform low-temperature electrical transport measurements on gated, quasi-2D graphite quantum dots. In devices with low contact resistances, we use longitudinal and Hall resistances to extract carrier densities of 9.2-13 x 10(12) cm(-2) and mobilities of 200-1900 cm(2)/V.s. In devices with high resistance contacts, we observe Coulomb blockade phenomena and infer the charging energies and capacitive couplings. These experiments demonstrate that electrons in mesoscopic graphite pieces are delocalized over nearly the whole graphite piece down to low temperatures. PMID- 15794613 TI - Stability and signatures of biexcitons in carbon nanotubes. AB - The linear optical properties of semiconducting carbon nanotubes are dominated by quasi-one-dimensional excitons formed by single electron-hole pairs. Hence, the nonlinear response at high pump levels most likely leads to the formation of exciton complexes involving several electron-hole pairs. Such complexes would therefore play an important role in, e.g., lasing applications. We demonstrate here that the biexciton complex is surprisingly stable for nanotubes in a wide diameter range. Theoretical predictions for the signature of such states in pump probe spectroscopy are presented. PMID- 15794614 TI - The preparation of submicrometer-sized rings by embedding and selective etching of spherical silica particles. AB - Silica particles were embedded in a polymer layer on a water surface in such a way that they protruded out of the top and the bottom surfaces. Etching of this layer with hydrofluoric acid vapors gradually removed the particles to yield membranes with pores that continuously widen until the particles are completely removed. Removing the polymer at intermediate stages of etching revealed that the particles are first converted into rings and sickles before they are finally completely destroyed. PMID- 15794615 TI - Gold nanorods as novel nonbleaching plasmon-based orientation sensors for polarized single-particle microscopy. AB - By monitoring the polarized light scattering from individual gold nanorods in a darkfield microscope, we are able to determine their orientation as a function of time. We demonstrate time resolution of milliseconds and observation times of hours by observing the two-dimensional rotational diffusion of gold rods attached to a glass surface. The observed orientational diffusion shows a fast component of about 60 ms and "sticky times" of seconds. The large signal-to-noise ratio, chemical and photochemical stability, fast time response, and small size of these gold nanorods make them an ideal probe for orientation sensing in material science and molecular biology. PMID- 15794616 TI - Mechanically adjustable and electrically gated single-molecule transistors. AB - We demonstrate a device geometry for single-molecule electronics experiments that combines both the ability to adjust the spacing between the electrodes mechanically and the ability to shift the energy levels in the molecule using a gate electrode. With the independent in-situ variations of molecular properties provided by these two experimental "knobs", we are able to achieve a much more detailed characterization of electron transport through the molecule than is possible with either technique separately. We illustrate the performance of the device using C(60) molecules. PMID- 15794617 TI - Two-photon photopolymerized tips for adhesion-free scanning-probe microscopy. AB - A hydrophobic polymeric tip of atomic force microscopy has been fabricated by two photon adsorbed photopolymerization methods (TPAP). The fabrication was performed by a layer-by-layer polymerization of sliced multiple three-dimensional computer aided design data. The used base resin was composed of acrylate and epoxy, which showed hydrophobic properties after the photopolymerization. For the sharp tip fabrication, we used a "dynamic partial polymerization method" which applied "the threshold effect" of TPAP to maximize the fabrication resolution. To investigate the performance of the fabricated polymeric tip, we have imaged several organic, inorganic, and biological samples using contact or dynamic force mode. The imaging results showed the hydrophobic polymeric tips solved various problems related to the tip adhesion to hydrophilic sample surfaces. Finally, the topographic image resolution of sub-5 nm was obtained using the polymeric tips for the hydrophilic mica surface. PMID- 15794618 TI - Stereocomplex block copolymer micelles: core-shell nanostructures with enhanced stability. AB - Monodisperse stereocomplex block copolymer micelles were obtained through the self-assembly of equimolar mixtures of poly(ethylene glycol)-block-poly(l lactide) and poly(ethylene glycol)-block-poly(d-lactide) in water. These micelles possessed partially crystallized cores and mean hydrodynamic diameters ranging from 31 to 56 nm, depending on the lactide content. They exhibited kinetic stability and redispersion properties superior to micelles prepared with isotactic or racemic polymers alone. This study demonstrates the advantages of stereocomplex formation in the design of stabilized water-soluble nanoparticles. PMID- 15794619 TI - Generating an etch resistant "resist" layer from common solvents using scanning probe lithography in a fluid cell. AB - A novel scanning probe lithography scheme is introduced involving the field induced deposition of etch resistant material generated from common organic solvents such as n-octane, toluene, ethyl alcohol, and dioxane in the tip/sample gap region of a liquid cell. An NH(4)F/H(2)O(2)/H(2)O etchant transfers these structures into 7 nm tall posts in a negative-tone fashion, indicating that an etch resistant, likely carbon-based material is produced by field-induced decomposition of the solvent. This is in sharp contrast to the positive tone images that result from a similar process involving water as the gap electrolyte followed by a similar fluorine-based etching. PMID- 15794620 TI - Super pH-sensitive multifunctional polymeric micelle. AB - To endow enhanced tumor specificity and endosome disruption property on the carrier, a multifunctional polymeric micelle was investigated. The micelle exposes the cell interacting ligand (biotin) on the surface under slightly acidic environmental conditions of various solid tumors and is internalized by biotin receptor-mediated endocytosis in a short time period. The micelle also showed pH dependent dissociation, causing the enhanced release of doxorubicin from the carrier in early endosomal pH. The dissociated micellar components subsequently disrupt endosomal membrane. PMID- 15794622 TI - Imperfect perfect lens. AB - We have quantitatively established a fundamental limitation on the ultimate spatial resolution of the perfect lens (thin metal slab) in the near field. This limitation stems from the spatial dispersion of the dielectric response of the Fermi liquid of electrons with Coulomb interaction in the metal. We discuss possible applications in nanoimaging, nanophotolithography, and nanospectroscopy. PMID- 15794621 TI - Cytotoxicity of colloidal CdSe and CdSe/ZnS nanoparticles. AB - Cytotoxicity of CdSe and CdSe/ZnS nanoparticles has been investigated for different surface modifications such as coating with mercaptopropionic acid, silanization, and polymer coating. For all cases, quantitative values for the onset of cytotoxic effects in serum-free culture media are given. These values are correlated with microscope images in which the uptake of the particles by the cells has been investigated. Our data suggest that in addition to the release of toxic Cd(2+) ions from the particles also their surface chemistry, in particular their stability toward aggregation, plays an important role for cytotoxic effects. Additional patch clamp experiments investigate effects of the particles on currents through ion channels. PMID- 15794624 TI - Prediction of ordered phases of encapsulated C60, C70, and C78 inside carbon nanotubes. AB - We report the first detailed fully atomistic molecular dynamics study of the encapsulation of symmetric (C(60)) and asymmetric fullerenes (C(70) and C(78)) inside single-walled carbon nanotubes of different diameters. Different ordered phases have been found and shown to be tube diameter dependent. Rotational structural disorder significantly affecting the volume fraction of the packing was observed for the molecular arrangements of asymmetric fullerenes. Although these effects make more difficult the existence of ordered phases, our results showed that complex packing arrangements (very similar to the ones obtained for C(60)) are also possible for C(70) and C(78). Comparisons with results from continuum and hard-sphere models, ab initio electronic structure calculations, and simulations of the high-resolution transmission electron microscopy images of the obtained fullerene packing phases are also presented. PMID- 15794623 TI - High performance n-type carbon nanotube field-effect transistors with chemically doped contacts. AB - Short channel ( approximately 80 nm) n-type single-walled carbon nanotube (SWNT) field-effect transistors (FETs) with potassium (K) doped source and drain regions and high-kappa gate dielectrics (ALD HfO(2)) are obtained. For nanotubes with diameter approximately 1.6 nm and band gap approximately 0.55 eV, we obtain n MOSFET-like devices exhibiting high on-currents due to chemically suppressed Schottky barriers at the contacts, subthreshold swing of 70 mV/decade, negligible ambipolar conduction, and high on/off ratios up to 10(6) at a bias voltage of 0.5 V. The results compare favorably with the state-of-the-art silicon n-MOSFETs and demonstrate the potential of SWNTs for future complementary electronics. The effects of doping level on the electrical characteristics of the nanotube devices are discussed. PMID- 15794625 TI - Controlled placement of CdSe nanoparticles in diblock copolymer templates by electrophoretic deposition. AB - An electrophoretic deposition process is shown to be an effective means of placing CdSe nanoparticles into nanopores and nanotrenches in templates prepared from polystyrene-block-poly(methyl methacrylate) (PS-b-PMMA) diblock copolymers. CdSe nanoparticles covered with alpha,omega-thiocarboxylic acid ligands were used in these studies, and an electric field was used to drive the nanoparticle deposition. Photoluminescence of the quantum dots was maintained within the nanostructures. PMID- 15794626 TI - A molecular dynamics study of the structural dependence of boron oxide nanoparticles on shape. AB - Molecular dynamics simulation is employed to study the effect of varying nanoparticle shape on the structure of boron oxide nanoparticles. Two nanoshapes are investigated and compared: a sphere of diameter 16 A and a cube of dimension 16 x 16 x 16 A. A many-body polarization model is employed within the simulation, accounting for dipole moments induced by local electric fields. The resulting network is described by a short-range structure consisting of planar BO(3) units, while the intermediate-range structure is described by six-membered planar boroxol rings. Both the fraction of boroxol rings and their locations differ between the two nanoshapes. All planar boroxol rings within the spherical simulation are located on the interior, while planar rings within the cubic simulation aggregate to the cube walls. In addition, structural differences appear between the two shapes at longer ranges, including the formation of "layers" aligned parallel to the walls of the cube, reminiscent of both the low density crystalline phase and the high-density amorphous form of boron oxide. PMID- 15794627 TI - Directed self-assembly of Ge nanostructures on very high index, highly anisotropic Si(hkl) surfaces. AB - Families of very high-index planes, such as those which bifurcate spontaneously to form a hill-and-valley structure composed of opposing facets, provide natural templates for the directed growth of position-controlled self-organized nanostructures with shapes determined by the facet width ratio R. For example, deposition of a few ML of Ge on Si(173 100 373), corresponding to R(113/517) = 1.7, results in a field of 40-nm-wide Ge nanowires along [72 187] with a uniform period of 60 nm. PMID- 15794628 TI - Synthesis of nearly monodisperse embedded nanoparticles by separating nucleation and growth in ion implantation. AB - We investigate the formation of nanoparticles of Au in SiO(2) by multiple ion implantation steps and intermediate anneals to isolate nucleation and growth, thereby producing a narrow particle size distribution. We discuss the effects of varying the initial nucleation dose and the growth temperature and establish guidelines for synthesizing nanoparticles with improved size uniformity. By this method, we have obtained a standard deviation of 16% on an average diameter of 1.68 nm, compared to 28% when no attempt is made to isolate nucleation and growth. PMID- 15794629 TI - Dumbbell-like bifunctional Au-Fe3O4 nanoparticles. AB - Dumbbell-like Au-Fe(3)O(4) nanoparticles are synthesized using decomposition of Fe(CO)(5) on the surface of the Au nanoparticles followed by oxidation in 1 octadecene solvent. The size of the particles is tuned from 2 to 8 nm for Au and 4 nm to 20 nm for Fe(3)O(4). The particles show the characteristic surface plasmon absorption of Au and the magnetic properties of Fe(3)O(4) that are affected by the interactions between Au and Fe(3)O(4). The dumbbell is formed through epitaxial growth of iron oxide on the Au seeds, and the growth can be affected by the polarity of the solvent, as the use of diphenyl ether results in flower-like Au-Fe(3)O(4) nanoparticles. PMID- 15794630 TI - Ultrafast dynamics of a ferromagnetic nanocomposite. AB - Ensembles of iron nanocrystals up to 25 nm in diameter embedded in SiO(2) were found to exhibit an ultrafast magnetic response to a transient out-of-plane magnetic field. The response time varies as a function of in-plane bias magnetic field with the fastest rise times, as short as 26 ps, observed for both zero and high bias fields (140 kA/m). Analytical modeling and micromagnetic simulations confirm that magnetostatic interactions between nanoparticles play an important role in the dynamic response. PMID- 15794631 TI - New and exotic self-organized patterns for modulated nanoscale systems. AB - The self-assembled domain patterns of modulated systems are the result of competing short-range attractive and long-range repulsive interactions found in diverse physical and chemical systems. From an application point of view, there is considerable interest in these domain patterns, as they form templates suitable for the fabrication of nanostructures. In this work we have generated a variety of new and exotic patterns, which represent either metastable or glassy states. These patterns arise as a compromise between the required equilibrium modulation period and the strain resulting from topologically constrained trajectories in phase space that effectively preclude the equilibrium configuration. PMID- 15794632 TI - Nanowires and nanoribbons of charge-density-wave conductor NbSe3. AB - We report synthesis of nanowires and nanoribbons of the charge-density-wave conductor NbSe(3) through direct reaction of Nb and Se powders. The transverse dimension of the obtained nanostructures, as identified with scanning/transmission electron microscopy, ranges from 20 to 700 nm. X-ray and selected area electron diffraction analyses indicate that these nanowires and nanoribbons are single crystalline. Four-probe resistivity measurements confirm the expected charge-density-wave transitions, and furthermore, we find significant enhancement in the depinning threshold fields, which we attribute to a confinement effect. PMID- 15794633 TI - Single DNA molecule detection using nanopipettes and nanoparticles. AB - Single DNA molecules labeled with nanoparticles can be detected by blockades of ionic current as they are translocated through a nanopipette tip formed by a pulled glass capillary. The nanopipette detection technique can provide not only tools for detection and identification of single DNA and protein molecules but also deeper insight and understanding of stochastic interactions of various biomolecules with their environment. PMID- 15794634 TI - RNA and protein folding: common themes and variations. AB - Visualizing the navigation of an ensemble of unfolded molecules through the bumpy energy landscape in search of the native state gives a pictorial view of biomolecular folding. This picture, when combined with concepts in polymer theory, provides a unified theory of RNA and protein folding. Just as for proteins, the major folding free energy barrier for RNA scales sublinearly with the number of nucleotides, which allows us to extract the elusive prefactor for RNA folding. Several folding scenarios can be anticipated by considering variations in the energy landscape that depend on sequence, native topology, and external conditions. RNA and protein folding mechanism can be described by the kinetic partitioning mechanism (KPM) according to which a fraction (Phi) of molecules reaches the native state directly, whereas the remaining fraction gets kinetically trapped in metastable conformations. For two-state folders Phi approximately 1. Molecular chaperones are recruited to assist protein folding whenever Phi is small. We show that the iterative annealing mechanism, introduced to describe chaperonin-mediated folding, can be generalized to understand protein assisted RNA folding. The major differences between the folding of proteins and RNA arise in the early stages of folding. For RNA, folding can only begin after the polyelectrolyte problem is solved, whereas protein collapse requires burial of hydrophobic residues. Cross-fertilization of ideas between the two fields should lead to an understanding of how RNA and proteins solve their folding problems. PMID- 15794635 TI - A nucleotide binding site in caspase-9 regulates apoptosome activation. AB - ATP or dATP is a required activator of Apaf-1 for formation of the Apoptosome and thereby activation of caspase-9 (Csp9) [Zou, H., Henzel, W. J., Liu, X., Lutschg, A., and Wang, X. (1997) Cell 90, 405-413]. Here we demonstrate that dATP or ATP may have an additional role in controlling Apaf-1-mediated Csp9 activation. In the presence of cytochrome c (CytC), dATP or ATP binds to Apaf-1 and triggers heptamerization of Apaf-1 leading to the activation of Csp9. At concentrations greater than 1 mM, dATP or ATP also functions as a negative regulator of apoptosis by binding to and inhibiting Csp9. The affinity labeling reagent, 3'-O (5-fluoro-2,4-dinitrophenyl)-ATP (FDNP-ATP), was used to probe the binding of nucleotides to Csp9. Similar to ATP, but with a much more profound effect, FDNP ATP binds to the full-length proCsp9 potently, with an IC(50) of approximately 5 11 nM. Neither ATP nor FDNP-ATP exhibits any effect on the prodomain-truncated enzyme DeltaproCsp9 or p18/p10. FDNP-ATP covalently labels proCsp9 with a stoichiometry of 1:1, resulting in DNP-ATP-proCsp9 that is incapable of forming a productive Apoptosome with Apaf-1. Activity assays show that ATP and dATP, but not ADP or AMP, bind to the processed Csp9 p35/p10. This nucleotide binding site might play an important and previously unrecognized role in regulating proCsp9 activation. PMID- 15794636 TI - Oxidative metabolites accelerate Alzheimer's amyloidogenesis by a two-step mechanism, eliminating the requirement for nucleation. AB - The process of amyloid formation by the amyloid beta peptide (Abeta), i.e., the misassembly of Abetapeptides into soluble quaternary structures and, ultimately, amyloid fibrils, appears to be at the center of Alzheimer's disease (AD) pathology. We have shown that abnormal oxidative metabolites, including cholesterol-derived aldehydes, modify Abeta and accelerate the early stages of amyloidogenesis (the formation of spherical aggregates). This process, which we have termed metabolite-initiated protein misfolding, could explain why hypercholesterolemia and inflammation are risk factors for sporadic AD. Herein, the mechanism by which cholesterol metabolites hasten Abeta 1-40 amyloidogenesis is explored, revealing a process that has at least two steps. In the first step, metabolites modify Abeta peptides by Schiff base formation. The Abeta-metabolite adducts form spherical aggregates by a downhill polymerization that does not require a nucleation step, dramatically accelerating Abeta aggregation. In agitated samples, a second step occurs in which fibrillar aggregates form, a step also accelerated by cholesterol metabolites. However, the metabolites do not affect the rate of fibril growth in seeded aggregation assays; their role appears to be in initiating amyloidogenesis by lowering the critical concentration for aggregation into the nanomolar range. Small molecules that block Schiff base formation inhibit the metabolite effect, demonstrating the importance of the covalent adduct. Metabolite-initiated amyloidogenesis offers an explanation for how Abeta aggregation could occur at physiological nanomolar concentrations. PMID- 15794639 TI - Structure of the conserved transcriptional repressor enhancer of rudimentary homolog. AB - erh (enhancer of rudimentary homolog) is a ubiquitously expressed transcriptional coregulator that is highly conserved among eukaryotes, from humans to plants to protozoa. Functions attributed to erh include enhancement of pyrimidine biosynthesis, a role in cell cycle regulation, and repression of the tissue specific transcription factor HNF-1 (hepatocyte nuclear factor-1) through binding the coactivator DCoH (dimerization cofactor of HNF1). No homologous sequences, other than erh orthologs, have been identified, and little is known about the interactions of erh. To further elucidate its function, we determined the crystal structure of erh to 2.0 A resolution. The erh structure is a novel alpha + beta fold consisting of a four-stranded antiparallel beta sheet with three amphipathic alpha helices situated on one face of the beta sheet. Structure-based searches of the Protein Data Bank, like sequence-based searches, failed to identify paralogs. We present structural and biochemical evidence that erh functions as a dimer. The dimer interface consists of a beta sandwich composed of the beta sheet from each monomer. Many of the surface residues of erh are conserved, including patches of hydrophobic and charged residues, suggesting protein-protein interaction interfaces. Two putative CKII phosphorylation sites are highly ordered in the structure and are predicted to disrupt dimerization and protein-protein interactions. PMID- 15794638 TI - Diabetes-associated mutations in human insulin: crystal structure and photo-cross linking studies of a-chain variant insulin Wakayama. AB - Naturally occurring mutations in insulin associated with diabetes mellitus identify critical determinants of its biological activity. Here, we describe the crystal structure of insulin Wakayama, a clinical variant in which a conserved valine in the A chain (residue A3) is substituted by leucine. The substitution occurs within a crevice adjoining the classical receptor-binding surface and impairs receptor binding by 500-fold, an unusually severe decrement among mutant insulins. To resolve whether such decreased activity is directly or indirectly mediated by the variant side chain, we have determined the crystal structure of Leu(A3)-insulin and investigated the photo-cross-linking properties of an A3 analogue containing p-azidophenylalanine. The structure, characterized in a novel crystal form as an R(6) zinc hexamer at 2.3 A resolution, is essentially identical to that of the wild-type R(6) hexamer. The variant side chain remains buried in a nativelike crevice with small adjustments in surrounding side chains. The corresponding photoactivatable analogue, although of low affinity, exhibits efficient cross-linking to the insulin receptor. The site of photo-cross-linking lies within a 14 kDa C-terminal domain of the alpha-subunit. This domain, unrelated in sequence to the major insulin-binding region in the N-terminal L1 beta-helix, is also contacted by photoactivatable probes at positions A8 and B25. Packing of Val(A3) at this interface may require a conformational change in the B chain to expose the A3-related crevice. The structure of insulin Wakayama thus evokes the reasoning of Sherlock Holmes in "the curious incident of the dog in the night": the apparent absence of structural perturbations (like the dog that did not bark) provides a critical clue to the function of a hidden receptor binding surface. PMID- 15794637 TI - Chiral mutagenesis of insulin. Foldability and function are inversely regulated by a stereospecific switch in the B chain. AB - How insulin binds to its receptor is unknown despite decades of investigation. Here, we employ chiral mutagenesis-comparison of corresponding d and l amino acid substitutions in the hormone-to define a structural switch between folding competent and active conformations. Our strategy is motivated by the T --> R transition, an allosteric feature of zinc-hexamer assembly in which an invariant glycine in the B chain changes conformations. In the classical T state, Gly(B8) lies within a beta-turn and exhibits a positive phi angle (like a d amino acid); in the alternative R state, Gly(B8) is part of an alpha-helix and exhibits a negative phi angle (like an l amino acid). Respective B chain libraries containing mixtures of d or l substitutions at B8 exhibit a stereospecific perturbation of insulin chain combination: l amino acids impede native disulfide pairing, whereas diverse d substitutions are well-tolerated. Strikingly, d substitutions at B8 enhance both synthetic yield and thermodynamic stability but markedly impair biological activity. The NMR structure of such an inactive analogue (as an engineered T-like monomer) is essentially identical to that of native insulin. By contrast, l analogues exhibit impaired folding and stability. Although synthetic yields are very low, such analogues can be highly active. Despite the profound differences between the foldabilities of d and l analogues, crystallization trials suggest that on protein assembly substitutions of either class can be accommodated within classical T or R states. Comparison between such diastereomeric analogues thus implies that the T state represents an inactive but folding-competent conformation. We propose that within folding intermediates the sign of the B8 phi angle exerts kinetic control in a rugged landscape to distinguish between trajectories associated with productive disulfide pairing (positive T-like values) or off-pathway events (negative R-like values). We further propose that the crystallographic T -->R transition in part recapitulates how the conformation of an insulin monomer changes on receptor binding. At the very least the ostensibly unrelated processes of disulfide pairing, allosteric assembly, and receptor binding appear to utilize the same residue as a structural switch; an "ambidextrous" glycine unhindered by the chiral restrictions of the Ramachandran plane. We speculate that this switch operates to protect insulin-and the beta-cell-from protein misfolding. PMID- 15794641 TI - Triple effect of mimetic peptides interfering with neural cell adhesion molecule homophilic cis interactions. AB - The neural cell adhesion molecule (NCAM) is pivotal in neural development, regeneration, and learning. Here we characterize two peptides, termed P1-B and P2, derived from the homophilic binding sites in the first two N-terminal immunoglobulin (Ig) modules of NCAM, with regard to their effects on neurite extension and adhesion. To evaluate how interference of these mimetic peptides with NCAM homophilic interactions in cis influences NCAM binding in trans, we employed a coculture system in which PC12-E2 cells were grown on monolayers of fibroblasts with or without NCAM expression and the rate of neurite outgrowth subsequently was analyzed. P2, but not P1-B, induced neurite outgrowth in the absence of NCAM binding in trans. When PC12-E2 cells were grown on monolayers of NCAM-expressing fibroblasts, the effect of both P1-B and P2 on neurite outgrowth was dependent on peptide concentrations. P1-B and P2 acted as conventional antagonists, agonists, and reverse agonists of NCAM at low, intermediate, and high peptide concentrations, respectively. The demonstrated in vitro triple pharmacological effect of mimetic peptides interfering with the NCAM homophilic cis binding will be valuable for the understanding of the actions of these mimetics in vivo. PMID- 15794640 TI - Differential DNA recognition and cleavage by EcoRI dependent on the dynamic equilibrium between the two forms of the malondialdehyde-deoxyguanosine adduct. AB - DNA damage may alter the outcome of protein-nucleic acid interactions. The malondialdehyde-deoxyguanosine adduct, 3-(2'-deoxy-beta-d-erythro pentofuranosyl)pyrimido[1,2-alpha]purin-10-(3H)-one (M(1)dG), miscodes in vivo and in vitro. M(1)dG is an exocyclic adduct that undergoes ring-opening in duplex DNA to form the acyclic adduct, N(2)-(3-oxo-1-propenyl)-deoxyguanosine (N(2) OPdG). These two adducts have different effects on DNA polymerase bypass and may affect other DNA processing enzymes. We employed the EcoRI restriction endonuclease as a model for the interaction of DNA binding proteins with adducted DNA substrates. The presence of M(1)dG in the EcoRI recognition sequence impaired the ability of the enzyme to cleave DNA, resulting in only 60% cleavage of the adducted strand and 75% cleavage of the complementary strand. Three adducts of similar structure to M(1)dG that are unable to ring-open were cleaved poorly, or not at all, by EcoRI. None of the adducts appeared to inactivate or sequester EcoRI. Additional studies with BssHII and PauI confirmed these results and demonstrated a positional effect of M(1)dG on cleavage efficiency. These data suggest dissimilar modes of protein-nucleic acid interactions based on differences in adduct structure. Comparison of the solution structures of DNA adducts and the crystal structure of EcoRI complexed to substrate suggest a model to explain the functional differences. PMID- 15794642 TI - Epigallocatechin gallate inhibits aryl hydrocarbon receptor gene transcription through an indirect mechanism involving binding to a 90 kDa heat shock protein. AB - The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor known to mediate the toxic effects of numerous environmental contaminants, including the polycyclic aromatic hydrocarbons (PAHs). Historically, binding of PAHs to the AhR and the events leading to the generation of DNA adducts have been associated with chemical carcinogenesis. Previous investigations have implicated green tea (GT) as affording protection against PAH-induced cancers in animal models. Investigations in our laboratory have demonstrated that the GT polyphenol epigallocatechin gallate (EGCG) is capable of antagonizing AhR-mediated gene transcription, implicating inhibition of AhR signaling as a potential chemopreventive mechanism. This line of investigation was directed at elucidating the molecular mechanism of this antagonism. Competitive binding assays strongly suggest that EGCG does not bind to the AhR ligand binding site, indicating this compound functions through a mechanism unlike that of typical AhR antagonists. Affinity chromatography experiments implicate an indirect mechanism of action involving direct binding of EGCG to the AhR chaperone protein, hsp90. This induces an AhR conformation capable of nuclear localization but incapable of binding DNA. These altered signaling events correlate with the formation of a complex with sedimentation characteristics different from those of the latent or ligand-activated AhR. These data implicate a model in which EGCG inhibits release of hsp90 from the AhR, stabilizing the complex in an intermediary state associated with XAP2. This is the first time EGCG has been demonstrated to directly bind hsp90 and the first indication that GT may exert its chemopreventive effects through an interaction with the common chaperone hsp90. PMID- 15794644 TI - NMR studies of restriction enzyme-DNA interactions: role of conformation in sequence specificity. AB - Sequence specific DNA binding proteins are thought to adopt distinct conformations when binding to target (cognate) and nontarget (noncognate) sequences. There is both biochemical and crystallographic evidence that this behavior is important in mediating sequence recognition by the Mg(II)-dependent type II restriction enzymes. Despite this, there are few systematic comparisons of the structural behavior of these enzymes in various complexes. Here, (1)H (15)N HSQC NMR spectroscopy is applied to PvuII endonuclease (2 x 18 kDa) in an effort to better understand the relationship between sequence recognition and enzyme conformational behavior. Spectra of the free enzyme collected in the absence and presence of metal ions indicate that while there is a modest backbone conformational response upon binding Ca(II), this does not occur with Mg(II). Substrate binding itself is accompanied by very dramatic spectral changes consistent with a large-scale conformational response. HSQC spectra of the enzyme bound to cognate (specific) and noncognate (nonspecific) oligonucleotides in the presence of Ca(II) are dramatically distinct, revealing for the first time the structural uniqueness of a PvuII cognate complex in solution. The strong correlation between NMR spectral overlap and crystallographic data (C(alpha) rmsd) permits characterization of the nonspecific PvuII complex as being more similar to the free enzyme than to the specific complex. Collectively, these data support the notion that it is the DNA, not the metal ion, which promotes a unique conformational response by the enzyme. It therefore follows that the principle role of metal ions in complex formation is one of driving substrate affinity and stability rather than conformationally priming the enzyme for substrate binding and sequence recognition. These results not only provide valuable insights into the mechanism of protein-DNA interactions but also demonstrate the utility of NMR spectroscopy in structure-function studies of these representative nucleic acid systems. PMID- 15794643 TI - Direct evidence for Sphingomonas sp. A1 periplasmic proteins as macromolecule binding proteins associated with the ABC transporter: molecular insights into alginate transport in the periplasm. AB - A Gram-negative bacterium, Sphingomonas sp. A1, has a macromolecule (alginate) import system consisting of a pit on the cell surface and an alginate-specific ATP-binding cassette importer in the inner membrane. Transport of alginate from the pit to the ABC importer is probably mediated by two periplasmic binding protein homologues (AlgQ1 and AlgQ2). Here we describe characteristics of binding of AlgQ1 and AlgQ2 to alginate and its oligosaccharides through surface plasmon resonance biosensor analysis, UV absorption difference spectroscopy, and X-ray crystallography. Both AlgQ1 and AlgQ2 were inducibly expressed in the periplasm of alginate-grown cells of strain A1. Biosensor analysis indicated that both proteins specifically bind alginate with a high degree of polymerization (>100) and that dissociation constants for alginate with an average molecular mass of 26 kDa are 2.3 x 10(-)(7) M for AlgQ1 and 1.5 x 10(-)(7) M for AlgQ2. An in vitro ATPase assay using the membrane complex, including the alginate ABC importer, suggested that both alginate-bound forms of AlgQ1 and AlgQ2 are closely associated with the importer. X-ray crystallography showed that AlgQ1 consisted of two domains separated by a deep cleft that binds alginate oligosaccharides through a conformational change in the two domains. These results directly show that alginate-binding proteins play an important role in the efficient transport of alginate macromolecules with different degrees of polymerization in the periplasm. PMID- 15794645 TI - Folding and binding integrity of variants of a prototype ligand-binding module from the LDL receptor possessing multiple alanine substitutions. AB - The LA repeats that comprise the ligand-binding domain of the LDL receptor are among the most common autonomously structured extracellular modules found in the nonredundant protein sequence database. Here, we investigate the information content of the amino acid sequence of a typical LA module by constructing sequences with alanine residues at nonconserved positions in the module. Starting with the sequence of the fifth ligand-binding repeat of the LDL receptor (LA5), we created generic LA modules with alanine substitutions of nonconserved residues in only the N-terminal lobe, only the C-terminal lobe, and throughout both lobes of the module. LA variants with alanine residues at as many as 18 of 37 positions fold to a preferred disulfide isomer in the presence of calcium. Indeed, the six cysteines, the C-terminal calcium coordinating residues, two hydrophobic residues involved in packing, two glycines, and five other residues that form side chain intramodule hydrogen bonds are alone sufficient to specify the fold of an LA module when alanine residues are present at all other positions. The LA variants with multiple alanines in either the N- or C-terminal lobe were then exploited to identify residues of LA5 that contribute to the binding of apoE-containing ligands in LDL receptor-derived "minireceptors", implicating nonconserved residues of the N-terminal lobe of LA5 in recognition of apoE-DMPC. Our library of LA modules with multiple alanine substitutions should be generally useful for probing the roles of nonconserved side chains in ligand recognition by proteins of the LDL receptor family. PMID- 15794646 TI - The role of Val-265 for flavin adenine dinucleotide (FAD) binding in pyruvate oxidase: FTIR, kinetic, and crystallographic studies on the enzyme variant V265A. AB - In pyruvate oxidase (POX) from Lactobacillus plantarum, valine 265 participates in binding the cofactor FAD and is responsible for the strained conformation of its isoalloxazine moiety that is visible in the crystal structure of POX. The contrasting effects of the conservative amino acid exchange V265A on the enzyme's catalytic properties, cofactor affinity, and protein structure were investigated. The most prominent effect of the exchange was observed in the 2.2 A crystal structure of the mutant POX. While the overall structures of the wild-type and the variant are similar, flavin binding in particular is clearly different. Local disorder at the isoalloxazine binding site prevents modeling of the complete FAD cofactor and two protein loops of the binding site. Only the ADP moiety shows well-defined electron density, indicating an "anchor" function for this part of the molecule. This notion is corroborated by competition experiments where ADP was used to displace FAD from the variant enzyme. Despite the fact that the affinity of FAD binding in the variant is reduced, the catalytic properties are very similar to the wild-type, and the redox potential of the bound flavin is the same for both proteins. The rate of electron transfer toward the flavin during turnover is reduced to one-third compared to the wild-type, but k(cat) remains unchanged. Redox-triggered FTIR difference spectroscopy of free FAD shows the nu(C(10a)=N(1)) band at 1548 cm(-)(1). In POX-V265A, this band is found at 1538 cm(-)(1) and thus shifted less strongly than in wild-type POX where it is found at 1534 cm(-)(1). Taking these observations together, the conservative exchange V265A in POX has a surprisingly small effect on the catalytic properties of the enzyme, whereas the effect on the three-dimensional structure is rather big. PMID- 15794647 TI - Ligand migration and protein fluctuations in myoglobin mutant L29W. AB - We have determined eight X-ray structures of myoglobin mutant L29W at various experimental conditions. In addition, infrared spectroscopic experiments are presented, which are discussed in the light of the X-ray structures. Two distinct conformations of the CO-ligated protein were identified, giving rise to two stretching bands of heme-bound CO. If L29W MbCO crystals are illuminated around 180 K, a deoxy species is formed. The CO molecules migrate to the proximal side of the heme and remain trapped in the so-called Xe1 cavity upon temperature decrease to 105 K. The structure of this photoproduct is almost identical to the equilibrium high-temperature deoxy Mb structure. If the temperature is cycled to increasingly higher values, CO recombination is observed. Three intermediate structures have been determined during the rebinding process. Efficient recombination occurs only above 180 K, the characteristic temperature for the onset of protein dynamics. Rebinding is remarkably slow because bulky residues His64 and Trp29 block important migration pathways of the CO molecule. PMID- 15794648 TI - Structural analysis of threonine 342 mutants of soybean beta-amylase: role of a conformational change of the inner loop in the catalytic mechanism. AB - Two different conformations of the inner loop (residues 340-346) have been found in the soybean beta-amylase structures. In the "product form", the Thr 342 residue creates hydrogen bonds with Glu 186 (catalytic acid) and with the glucose residues at subsites -1 and +1, whereas most of those interactions are lost in the "apo form". To elucidate the relationship between the structural states of the inner loop and the catalytic mechanism, Thr 342 was mutated to Val, Ser, and Ala, respectively, and their crystal structures complexed with maltose were determined together with that of the apo enzyme at 1.27-1.64 A resolutions. The k(cat) values of the T342V, T342S, and T342A mutants decreased by 13-, 360-, and 1700-fold, respectively, compared to that of the wild-type enzyme. Whereas the inner loops in the wild-type/maltose and T342V/maltose complexes adopted the product form, those of the T342S/maltose and T342A/maltose complexes showed the apo form. Structural analyses suggested that the side chain of Thr 342 in product form plays an important role in distorting the sugar ring at subsite -1, stabilizing the deprotonated form of Glu 186, and grasping the glucose residue of the remaining substrate at subsite +1. The third hypothesis was proved by the fact that T342V hydrolyzes maltoheptaose following only multichain attack in contrast to multiple attack of the wild-type enzyme. PMID- 15794649 TI - Effect of sterol regulatory element binding protein-1a on the mitochondrial protein pattern in human liver cells detected by 2D-DIGE. AB - Sterol regulatory element binding protein-1a (SREBP-1a) is a transcription factor that is a major player in lipid metabolism and insulin action. We have generated human liver cells (HepG2) overexpressing active SREBP-1a constitutively called SREBP-1a (+). These cells show massive intracellular lipid accumulation. To elucidate the effect of SREBP-1a on lipid metabolism at the level of the cellular protein network, we have analyzed the protein pattern of mitochondria using the novel technique two-dimensional difference gel electrophoresis (2D-DIGE). Mitochondria were enriched by subcellular fractionation using differential and isopyknic centrifugation. Proteins of isolated organelles were labeled with Cy dyes and separated on 2D gels. These gels revealed more than 100 protein spots, which were significantly different in their abundance between wild-type and SREBP 1a (+) cells. MALDI mass spectrometry showed that 68% of identified proteins belong to mitochondria. In SREBP-1a (+) cells, several enzymes involved in lipid metabolism were significantly altered, suggesting that cellular lipid metabolism is triggered by accumulation of fatty acids rather than by its degradation. To test the possible functional relevance of this finding, intracellular fatty acid (FA) patterns were analyzed by gas chromatography. The results showed a significant increase in total fatty acid content with a shift in composition to long-chain unsaturated FAs. Therefore, the detected protein differences might be an explanation for the observed intracellular lipid accumulation and might link SREBP-1a to features like steatosis hepatis. PMID- 15794651 TI - Purification and physical-chemical properties of methanobactin: a chalkophore from Methylosinus trichosporium OB3b. AB - Methanobactin is an extracellular, copper-binding chromopeptide from the methane oxidizing bacterium, Methylosinus trichosporium OB3b, believed to be involved in copper detoxification, sequestration, and uptake. Although small (1217.2 Da), methanobactin possesses a complex three-dimensional macrocyclic structure with several unusual moieties. The molecule binds one copper and has the N-2 isopropylester-(4-thionyl-5-hydroxyimidazolate)-Gly(1)-Ser(2)-Cys(3)-Tyr(4) pyrrolidine-(4-hydroxy-5-thionylimidazolate)-Ser(5)-Cys(6)-Met(7) sequence [Kim, H. J., et al. (2004) Science 305, 1612-1615]. We report methods for purifying methanobactin from M. trichosporium OB3b and present initial evidence of its physiological function. MALDI-TOF MS was used to systematically monitor samples for optimizing purification conditions, and for detecting and analyzing specific metal-methanobactin complexes. Purification was performed by first stabilizing the extracted compound with copper followed by separation using reversed-phase HPLC in neutral pH buffers. Purified methanobactin exhibited UV-visible maxima at 342 nm, a shoulder at 388 nm, and a broad peak at 282 nm. These features were lost upon CuCl(2) titration with appearance of new features at 335, 356, 290, and 255 nm. Furthermore, methanobactin contains two fluorescent moieties, which exhibit broad emissions at 440-460 nm (lambda(max)(ex) at 388 nm) and 390-430 nm (lambda(max)(ex) = 342 nm), respectively. Finally, methanobactin eliminates the growth lag in M. trichosporium OB3b and substantially increases growth rates when cultures are exposed to elevated copper levels. PMID- 15794650 TI - Design and expression of cysteine-bearing hydrophobic polypeptides and their self assembling properties with bacteriochlorophyll a derivatives as a mimic of bacterial photosynthetic antenna complexes. Effect of steric confinement and orientation of the polypeptides on the pigment/polypeptide assembly process. AB - A series of cysteine-bearing hydrophobic polypeptides analogous to a light harvesting one betapolypeptide (LH1beta) from the LH1 complex from the purple photosynthetic bacterium, Rhodobacter sphaeroides, was synthesized using an Escherichia coli expression system. The cysteine was placed in the C- or N terminal regions of the polypeptide to investigate the influence of steric confinement and orientation of the polypeptides via disulfide linkages as they were self-assembled with zinc-substituted bacteriochlorophyll a ([Zn]-BChl a). The polypeptides were expressed as water-soluble fusion proteins with maltose binding protein (MBP). The fusion proteins formed a subunit-type complex with the [Zn]-BChl a in an n-octyl-beta-d-glucopyranoside (OG) micellar solution regardless of the cross-links or the cleavage of the cysteines, judging from absorption, CD, and fluorescence spectra. Following treatment with trypsin, the polypeptides were detached from the MBP portion. Such trypsin-digested polypeptides formed a subunit-type LH complex at 25 degrees C, which also showed that the disulfide linkage was not crucial for the subunit formation. When a polypeptide having cysteine on the C-terminus was assembled at 4 degrees C, the Qy absorption band was remarkably red-shifted to approximately 836 nm, suggesting that the cleavage of the large MBP portion liberates the polypeptides to form the progressive type of complex similar to LH1-type complex. The trypsin-treated polypeptides bearing cysteines in both terminal regions, which are randomly cross linked, did not form the LH1-type complex under oxidative conditions but did form the complex under reductive conditions. This observation suggests that the polypeptide orientation strongly influences the LH1-type complex formation. The progressive assembly from the subunit to the holo-LH1-type complex following cleavage of MBP portion in a lipid bilayer is also briefly discussed. PMID- 15794652 TI - Primary intermediate in the photocycle of a blue-light sensory BLUF FAD-protein, Tll0078, of Thermosynechococcus elongatus BP-1. AB - Proteins with a BLUF (sensor of blue light using flavin adenine dinucleotide) domain represent a newly recognized class of photoreceptors that is widely distributed in the genomes of photosynthetic bacteria, cyanobacteria, and Euglena. Recently, Okajima et al. [Okajima, K., Yoshihara, S., Geng, X., Katayama, M. and Ikeuchi, M. (2003) Plant Cell Physiol. 44 (Suppl), 162] purified BLUF protein Tll0078 encoded in the genome of thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 by expressing the protein in Escherichia coli. We investigated the photocycle of Tll0078 by measuring the picosecond fluorescence kinetics, transient absorption changes, and the UV-visible absorption spectra at 10 to 330 K. The absorption spectrum of the FAD moiety of Tll0078 showed a 10-nm red shift upon illumination at 278-330 K. The quantum efficiency of the formation of the red-shifted form was 29%. Illumination at 10 K, on the other hand, caused only a 5-nm red shift in about one-half of the protein population. The 5-nm-shifted form was stable at 10 K. The 5-nm red shifted form was converted into the 10-nm red-shifted form at 50-240 K upon warming in the dark. At room temperature, the 10-nm red-shifted final product appeared within 10 ns after laser flash excitation. The lifetime of the fluorescence of FAD was found to be 120 ps at room temperature. These results reveal a fast and efficient photoconversion process from the singlet-excited state to the final product at room temperature. A photocycle of BLUF protein is proposed that includes the 5-nm red-shifted intermediate form as the precursor for the 10-nm red-shifted final product. The temperature dependence of each step of the photocycle is also discussed. PMID- 15794653 TI - Metal-binding characteristics of the amino-terminal domain of ZntA: binding of lead is different compared to cadmium and zinc. AB - ZntA from Escherichia coli, a P1-type ATPase, specifically transports Pb(II), Zn(II), and Cd(II). Most P1-type ATPases have an N-terminal domain that contains one or more copies of the conserved metal-binding motif, GXXCXXC. In ZntA, the N terminal domain has approximately 120 residues with a single GXXCXXC motif, as well as four additional cysteine residues as part of the CCCDGAC motif. The metal binding specificity and affinity of this domain in ZntA was investigated. Isolated proteins, N1-ZntA and N2-ZntA, containing residues 1-111 and 47-111 of ZntA, respectively, were characterized. N1-ZntA has both the CCCDGAC and GXXCXXC motifs, while N2-ZntA has only the GXXCXXC motif. ICP-MS measurements showed that N1-ZntA can bind both divalent metal ions such as Cd(II), Pb(II), and Zn(II) and monovalent metal ions such as Ag(I), with a stoichiometry of 1. N2-ZntA can bind Zn(II) and Cd(II) with a stoichiometry of 1 but not Pb(II). The affinity of N1 ZntA for Zn(II), Pb(II), and Cd(II) was measured by competition titration with metallochromic indicators. Association constants of approximately 10(8) M(-)(1) were obtained for Zn(II), Pb(II), and Cd(II) binding to N1-ZntA. To investigate whether the CCCDGAC sequence has an important role in binding specifically Pb(II), a mutant of ZntA, which lacked the first 46 residues, was constructed. This mutant, Delta46-ZntA, had the same activity as wtZntA with respect to Cd(II) and Zn(II). However, its activity with Pb(II) was similar to the mutant DeltaN ZntA, which lacks the entire N-terminal domain (Mitra, B., and Sharma, R. (2001) Biochemistry 40, 7694-7699). Thus, binding of Pb(II) appears to involve different ligands, and possibly geometry, compared to Cd(II) and Zn(II). PMID- 15794654 TI - Spectroscopic studies on cobalt(II)-substituted metallo-beta-lactamase ImiS from Aeromonas veronii bv. sobria. AB - In an effort to probe the structure of a group Bb metallo-beta-lactamase, Co(II) substituted ImiS was prepared and characterized by electronic absorption, NMR, and EPR spectroscopies. ImiS containing 1 equiv of Co(II) (Co(II)(1)-ImiS) was shown to be catalytically active. Electronic absorption studies of Co(II)(1)-ImiS revealed the presence of two distinct features: (1) an intense sulfur to Co(II) ligand to metal charge transfer band and (2) less intense, Co(II) ligand field transitions that suggest 4-coordinate Co(II) in Co(II)(1)-ImiS. (1)H NMR studies of Co(II)(1)-ImiS suggest that one histidine, one aspartic acid, and one cysteine coordinate the metal ion in Co(II)(1)-ImiS. The addition of a second Co(II) to Co(II)(1)-ImiS did not result in any additional solvent-exchangeable NMR resonances, strongly suggesting that the second Co(II) does not bind to a site with histidine ligands. EPR studies reveal that the metal ion in Co(II)(1)-ImiS is 4-coordinate and that the second Co(II) is 5/6 coordinate. Taken together, these data indicate that the catalytic site in ImiS is the consensus Zn(2) site, in which Co(II) (and by extrapolation Zn(II)) is 4-coordinate and bound by Cys221, His263, Asp120, and probably one solvent water molecule. These studies also show that the second, inhibitory metal ion does not bind to the consensus Zn(1) site and that the metal ion binds at a site significantly removed from the active site. These results give the first structural information on metallo-beta lactamase ImiS and suggest that the second metal binding site in ImiS may be targeted for inhibitors. PMID- 15794655 TI - Use of viscogens, dNTPalphaS, and rhodium(III) as probes in stopped-flow experiments to obtain new evidence for the mechanism of catalysis by DNA polymerase beta. AB - The kinetic mechanism and the structural bases of the fidelity of DNA polymerases are still highly controversial. Here we report the use of three probes in the stopped-flow studies of Pol beta to obtain new, direct evidence for our previous interpretations: (a) Increasing the viscosity of the reaction buffer by sucrose or glycerol is expected to slow down the conformational change differentially, and it was shown to slow down the first (fast) fluorescence transition selectively. (b) Use of dNTPalphaS in place of dNTP is expected to slow down the chemical step preferentially, and it was shown to slow down the second (slow) fluorescence transition selectively. (c) The substitution-inert Rh(III)dNTP was used to show for the first time that the slow fluorescence change occurs after mixing of Pol beta.DNA.Rh(III)dNTP with Mg(II). These results, along with crystal structures, suggest that the subdomain-closing conformational change occurs before binding of the catalytic Mg(II) while the rate-limiting step occurs after binding of the catalytic Mg(II). These results provide new evidence to the mechanism we suggested previously, but do not support the results of three recent papers of computational studies. The results were further supported by a "sequential mixing" stopped-flow experiment that used no analogues, and thus ruled out the possibility that the discrepancy between experimental and computational results is due to the use of analogues. The methodologies can be used to examine other DNA polymerases to answer whether the properties of Pol beta are exceptional or general. PMID- 15794656 TI - Controlling association of vesicle embedded peptides by alteration of the physical state of the lipid matrix. AB - We report here the reversible association of a designed peptide embedded in a lipid membrane through a stimulus-sensitive trigger that changes the physical state of the bilayer matrix. A peptide designed with the classical 4-3 heptad repeat of coiled coils, equipped with leucine residues at all canonical interface positions, TH1, was rendered membrane soluble by replacement of all exterior residues with randomly selected hydrophobic amino acids. Insertion of TH1 into large unilamellar phosphatidylcholine vesicles was followed by monitoring tryptophan fluorescence. Peptide insertion was observed when the lipids were in the liquid-crystalline state [1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)] but not when they were in the crystalline phase [1,2-dipalmitoyl-sn glycero-3-phosphocholine (DPPC)]. Formation of a trimeric alpha-helical bundle in lipid bilayers was followed by fluorescence resonance energy transfer. Global fit analysis revealed a monomer--trimer equilibrium with a dissociation constant of around 10(-5) [corrected] MF(2). A lipid mixture composed of DPPC and POPC exhibiting a phase transition at 34 degrees C between a crystalline/liquid crystalline coexistence region and a completely miscible liquid-crystalline phase was used to control the formation of the trimeric peptide bundle. TH1 is phase excluded in crystalline DPPC domains below 34 degrees C, leading to a larger number of trimers. However, when the DPPC domains are dispersed at temperatures above 34 degrees C, the number of trimers is reduced. PMID- 15794657 TI - NMR structure determination of a membrane protein with two transmembrane helices in micelles: MerF of the bacterial mercury detoxification system. AB - The three-dimensional backbone structure of a membrane protein with two transmembrane helices in micelles was determined using solution NMR methods that rely on the measurement of backbone (1)H-(15)N residual dipolar couplings (RDCs) from samples of two different constructs that align differently in stressed polyacrylamide gels. Dipolar wave fitting to the (1)H-(15)N RDCs determines the helical boundaries based on periodicity and was utilized in the generation of supplemental dihedral restraints for the helical segments. The (1)H-(15)N RDCs and supplemental dihedral restraints enable the determination of the structure of the helix-loop-helix core domain of the mercury transport membrane protein MerF with a backbone RMSD of 0.58 A. Moreover, the fold of this polypeptide demonstrates that the two vicinal pairs of cysteine residues, shown to be involved in the transport of Hg(II) across the membrane, are exposed to the cytoplasm. This finding differs from earlier structural and mechanistic models that were based primarily on the somewhat atypical hydropathy plot for MerF and related transport proteins. PMID- 15794658 TI - An alternate pattern for globoside oligosaccharide expression in Haemophilus influenzae lipopolysaccharide: structural diversity in nontypeable strain 1124. AB - Common structural motifs of Haemophilus influenzae lipopolysaccharide (LPS) are globotetraose [beta-d-GalpNAc-(1-->3)-alpha-d-Galp-(1-->4)-beta-d-Galp-(1-->4) beta-d-Glcp] and its truncated versions globoside [alpha-d-Galp-(1-->4)-beta-d Galp-(1-->4)-beta-d-Glcp] and lactose [beta-d-Galp-(1-->4)-beta-d-Glcp] linked to the terminal heptose (HepIII) of the triheptosyl inner-core moiety l-alpha-d-Hepp (1-->2)-[PEA-->6]-l-alpha-d-Hepp-(1-->3)-l-alpha-d-Hepp-(1-->5)-[PPEA-->4]-alpha Kdo-(2-->6)-lipid A. We report here structural studies of LPS from nontypeable H. influenzae strain 1124 expressing these motifs linked to both the proximal heptose (HepI) and HepIII at the same time. This novel finding was obtained by structural studies of LPS using NMR techniques and electrospray ionization mass spectrometry (ESI-MS) on O-deacylated LPS and core oligosaccharide material (OS) as well as ESI-MS(n)() on permethylated dephosphorylated OS. The use of defined mutants allowed us to confirm structures unambiguously and understand better the biosynthesis of each of the globotetraose units. We found that lgtC is involved in the expression of alpha-d-Galp-(1-->4)-beta-d-Galp in both extensions, whereas lic2A directs only the expression of beta-d-Galp-(1-->4)-beta-d-Glcp when linked to HepIII. The LPS of NTHi strain 1124 contained sialylated glycoforms that were identified by CE-ESI-MS/MS. A common sialylated structure in H. influenzae LPS is sialyllactose linked to HepIII. This structure exists in strain 1124. However, results for the lpsA mutant indicate that sialyllactose extends from HepI as well, a molecular environment for sialyllactose in H. influenzae that has not been reported previously. In addition, the LPS was found to carry phosphorylcholine, O-linked glycine, and a third PEA group which was linked to O3 of HepIII. PMID- 15794659 TI - Suppression of axial methionine fluxion in Hydrogenobacter thermophilus Gln64Asn cytochrome c552. AB - Proteins in the cytochrome c (cyt c) family with His-Met heme axial ligation display diverse heme electronic structures as revealed by the NMR spectra of their oxidized (paramagnetic) forms. These variations in electronic structure are thought to result primarily from differences in heme axial Met orientation among cyt c species. The factors determining Met orientation in cyts c, however, remain poorly understood. An additional layer of complexity was revealed with the recent finding that the axial Met in Hydrogenobacter thermophilus cytochrome c(552) (Ht cyt c(552)) is fluxional, sampling two conformations rapidly on the NMR time scale, resulting in an unusual compressed range of heme substituent hyperfine shifts [Zhong, L., Wen, X., Rabinowitz, T. M., Russell, B. S., Karan, E. F., and Bren, K. L. (2004) Proc.Natl. Acad. Sci. U.S.A. 101, 8637-8642]. In this work, the (1)H NMR hyperfine shift pattern of Ht cyt c(552) is drastically altered by making the conservative heme pocket mutation Gln64Asn. The mutant (Ht Q64N) displays a pattern of heme hyperfine shifts with a remarkable resemblance to that of structurally homologous Pseudomonas aeruginosa cyt c(551), which has Asn at position 64 and a single heme axial Met conformation. NMR analysis reveals that Asn64 in Ht Q64N is positioned to interact with the axial Met61, whereas the Gln64 in wild-type Ht cyt c(552) is not. It also is found that the heme axial Met is not fluxional in Ht Q64N and has an orientation similar to that in P. aeruginosa cyt c(551). These results indicate that peripheral interactions with the axial Met play an important role in determining axial Met orientation and heme electronic structure in cyts c. PMID- 15794660 TI - Accumulation of long-chain acylcarnitine and 3-hydroxy acylcarnitine molecular species in diabetic myocardium: identification of alterations in mitochondrial fatty acid processing in diabetic myocardium by shotgun lipidomics. AB - Diabetic cardiomyopathy is the result of maladaptive changes in energy homeostasis. However, the biochemical mechanisms underlying dysfunctional lipid metabolism in diabetic myocardium are incompletely understood. Herein, we exploit shotgun lipidomics to demonstrate a 4-fold increase in acylcarnitines in diabetic myocardium, which was reversible upon insulin treatment. Analysis of acylcarnitine molecular species in myocardium unexpectedly identified acylcarnitine molecular species containing a mass shift of 16 amu in comparison to the anticipated molecular species. Synthesis of 3-hydroxy acylcarnitine identified the natural products as the 3-hydroxylated acylcarnitines through comparisons of diagnostic fragmentation patterns of synthetic and naturally occurring constituents using tandem mass spectrometry. Diabetes induced an increase of both calcium-independent phospholipase A(2) (iPLA(2)) mRNA and iPLA(2) activity in rat myocardium. Cardiac ischemia in myocardium genetically engineered to overexpress iPLA(2) dramatically increased the amount of acylcarnitine present in myocardium. Moreover, mechanism-based inactivation of iPLA(2) in either wild-type or transgenic myocardium ablated a substantial portion of the acylcarnitine increase. Collectively, these results identify discrete insulin remediable abnormalities in mitochondrial fatty acid processing in diabetic myocardium and identify iPLA(2) as an important enzymatic contributor to the pool of fatty acids that can be used for acylcarnitine synthesis and energy production in myocardium. PMID- 15794661 TI - Proteolytic activation of protein kinase Calpha by peroxynitrite in stimulating cytosolic phospholipase A2 in pulmonary endothelium: involvement of a pertussis toxin sensitive protein. AB - We sought to determine the roles of PKCalpha and G(i)alpha in regulating cPLA(2) activity in bovine pulmonary artery endothelial cell membrane under peroxynitrite (ONOO(-)) stimulation. Treatment of bovine pulmonary artery endothelial cells with ONOO(-) markedly stimulates the cell membrane associated protease activity, protein kinase C (PKC) activity, phospholipase A(2) (PLA(2)) activity, and arachidonic acid (AA) release from the cells. ONOO(-) significantly increases (Ca(2+))(i) in the cells, and pretreatment with the intracellular Ca(2+) chelator BAPTA-AM prevents the increase in (Ca(2+))(i), protease activity, PKC activity, and cPLA(2) activity in the cell membrane and AA release from the cells. Pretreatment of the cells with arachidonyl trifluoromethyl ketone (AACOCF(3)) (a cPLA(2) inhibitor) prevents ONOO(-)-stimulated cPLA(2) activity and AA release without producing a significant alteration of the protease activity. Pretreatment with vitamin E and aprotinin prevents ONOO(-)-induced increase in the protease activity, PKC activity, and cPLA(2) activity in the cell membrane and AA release from the cells. Pretreatment with the PKC inhibitor calphostin C prevents ONOO(-) caused increase in PKC activity and cPLA(2) activity in the cell membrane and AA release from the cells. An immunoblot study of the cell membrane isolated from the ONOO(-)-treated cells with polyclonal PKCalpha antibody elicited an increase in the 80 kDa immunoreactive protein band along with an additional 47 kDa immunoreactive fragment. An immunoblot study with anti-nitrotyrosine antibody revealed that ONOO(-) induces nitration of tyrosine residues in PKCalpha. Pretreatment of the cells with aprotinin abolished the 47 kDa immunoreactive fragment in the immunoblot. An immunoblot study of the endothelial cell membrane with polyclonal cPLA(2) antibody revealed that treatment of the cells with ONOO( ) markedly increases the cPLA(2) immunoreactive protein profile in the membrane. Pretreatment of the endothelial cells with Go6976, a PKCalpha inhibitor, prevents the increase in PKC activity and cPLA(2) activity in the cell membrane under ONOO(-)-triggered condition. It, therefore, appears from the present study that treatment of the cells with ONOO(-) causes an increase in the protease activity, and that plays an important role in activating PKCalpha, which subsequently stimulates cPLA(2) activity in the cell membrane and AA release from the cells. An immunoblot assay with polyclonal G(i)alpha antibody elicited an immunoreactive band having a molecular mass of 41 kDa. Pretreatment of the cells with pertussis toxin markedly inhibits ONOO(-)-induced increase in cPLA(2) activity and AA release without significantly altering (Ca(2+))(i), protease activity, and PKC activity in the cell membrane. Treatment of the cells with ONOO(-) causes phosphorylation of G(i)alpha in the cell membrane, and pretreatment with Go6976 prevents its phosphorylation. We suggest the existence of a pertusssis toxin sensitive G protein-mediated mechanism for activation of cPLA(2) by ONOO(-) in bovine pulmonary artery endothelial cell membrane, which is regulated by PKCalpha dependent phosphorylation and sensitive to aprotinin for its inhibition. PMID- 15794662 TI - Thermodynamic stability of carbonic anhydrase: measurements of binding affinity and stoichiometry using ThermoFluor. AB - ThermoFluor (a miniaturized high-throughput protein stability assay) was used to analyze the linkage between protein thermal stability and ligand binding. Equilibrium binding ligands increase protein thermal stability by an amount proportional to the concentration and affinity of the ligand. Binding constants (K(b)) were measured by examining the systematic effect of ligand concentration on protein stability. The precise ligand effects depend on the thermodynamics of protein stability: in particular, the unfolding enthalpy. An extension of current theoretical treatments was developed for tight binding inhibitors, where ligand effect on T(m) can also reveal binding stoichiometry. A thermodynamic analysis of carbonic anhydrase by differential scanning calorimetry (DSC) enabled a dissection of the Gibbs free energy of stability into enthalpic and entropic components. Under certain conditions, thermal stability increased by over 30 degrees C; the heat capacity of protein unfolding was estimated from the dependence of calorimetric enthalpy on T(m). The binding affinity of six sulfonamide inhibitors to two isozymes (human type 1 and bovine type 2) was analyzed by both ThermoFluor and isothermal titration calorimetry (ITC), resulting in a good correlation in the rank ordering of ligand affinity. This combined investigation by ThermoFluor, ITC, and DSC provides a detailed picture of the linkage between ligand binding and protein stability. The systematic effect of ligands on stability is shown to be a general tool to measure affinity. PMID- 15794663 TI - Evaluation of add-on testing in the clinical chemistry laboratory of a large academic medical center. PMID- 15794665 TI - Pseudohypoglycemia with monoclonal immunoglobulin m. PMID- 15794667 TI - Patient safety in anatomic pathology: measuring discrepancy frequencies and causes. AB - CONTEXT: Anatomic pathology discrepancy frequencies have not been rigorously studied. OBJECTIVE: To determine the frequency of anatomic pathology discrepancies and the causes of these discrepancies. DESIGN: Participants in the College of American Pathologists Q-Probes program self-reported the number of anatomic pathology discrepancies in their laboratories by prospectively performing secondary review (post-sign-out) of 100 surgical pathology or cytology specimens. Reasons for the secondary review included conferences, external review, internal quality assurance policy, and physician request. PARTICIPANTS: Seventy-four laboratories self-reported data. MAIN OUTCOME MEASURES: Frequency of anatomic pathology discrepancy; type of discrepancy (ie, change in margin status, change in diagnosis, change in patient information, or typographic error); effect of discrepancy on patient outcome (ie, no harm, near miss, or harm); and clarity of report. RESULTS: The mean and median laboratory discrepancy frequencies were 6.7% and 5.1%, respectively. Forty-eight percent of all discrepancies were due to a change within the same category of interpretation (eg, 1 tumor type was changed to another tumor type). Twenty-one percent of all discrepancies were due to a change across categories of interpretation (eg, a benign diagnosis was changed to a malignant diagnosis). Although the majority of discrepancies had no effect on patient care, 5.3% had a moderate or marked effect on patient care.Conclusions. This study establishes a mean multi-institutional discrepancy frequency (related to secondary review) of 6.7%. PMID- 15794668 TI - Staffing benchmarks for clinical laboratories: a College of American Pathologists Q-probes study of staffing at 151 institutions. AB - CONTEXT: Inadequate staffing of clinical laboratories may compromise quality and throughput, whereas excess staff unnecessarily increases the cost of testing. OBJECTIVES: To measure productivity of technical staff and management span of control in a large number of laboratories and to determine factors associated with favorable staffing ratios. DESIGN: A total of 151 clinical laboratories provided information about technical and management staffing and output (workload) for 4 laboratory sections: anatomic pathology, chemistry/hematology/immunology, microbiology, and transfusion medicine. RESULTS: For each laboratory section, there was wide variation in labor productivity (output per nonmanagement full-time equivalent) and in management span of control (nonmanagement full-time equivalent per manager). Productivity ratios for the 10th- and 90th-percentile laboratories varied more than 3-fold. Except in histology, laboratory sections with higher test volumes had higher labor productivity (P < .001 for cytology, chemistry/hematology, and transfusion medicine; P = .003 for microbiology). Even within peer groups composed of sections with similar volume, there was wide variation in labor productivity. A number of variables other than test volume were associated with labor productivity and management span of control. Staffing ratios for each laboratory section and for sections of different sizes are presented. CONCLUSIONS: Despite standardization of testing methods in the clinical laboratory industry, there is wide variation in staffing level among institutions. This variation suggests opportunities to improve staff productivity in many facilities. PMID- 15794669 TI - Association of mild transient elevation of troponin I levels with increased mortality and major cardiovascular events in the general patient population. AB - CONTEXT: The prognostic value of mild elevation of cardiac-specific troponin I (cTnI) levels is poorly defined, which can make interpretation of such an elevation difficult. OBJECTIVE: To study the prognostic value of transient mild elevation of cTnI levels in the hospitalized patient population. DESIGN: We performed a case-control study that compared the outcome of patients hospitalized for any cause with at least 2 subsequent transient cTnI measurements of 0.1 ng/mL or higher and less than 1.5 ng/mL with matched controls with cTnI levels less than 0.1 ng/mL. A cohort of 118 patients (mean +/- SD age, 67.4 +/- 14.0 years; 35.6% men) was followed up for an average +/- SD of 11.9 +/- 7.9 months. Seventy one cases were matched with 37 controls in terms of demographics, coronary artery disease risk factors, and reason for admission. End points were all-cause mortality and major cardiovascular end points, including cardiovascular mortality, myocardial infarction, and revascularization. RESULTS: The total event rate was significantly increased in the case group compared with the control group at 12, 6, and 3 months (62.0% vs 24.3%, 59.2% vs 16.2%, and 47.9% vs 5.4%, respectively; P < .001). At 12, 6, and 3 months, the cases had a significant increase in all-cause mortality (43.7% vs 16.2%, 40.8% vs 8.1%, and 33.8% vs 0.0%, respectively; P = .005) and major cardiovascular end points (26.8% vs 8.1%, 26.8% vs 8.1%, and 21.1% vs 5.4%, respectively; P = .02) compared with controls. CONCLUSION: Transient mild elevation of cTnI levels in hospitalized patients is associated with an increase in all-cause mortality and major cardiovascular complications. Such elevations of cTnI levels can be considered a marker for both all-cause and cardiovascular morbidity and mortality. PMID- 15794670 TI - Regulators of apoptosis in cholangiocarcinoma. AB - CONTEXT: Dysregulation of mediators of apoptosis is associated with carcinogenesis. For biliary duct cancers, p53 gene mutation is an important contributor to carcinogenesis. Mutations in the p53 gene affect transcription of the Fas gene, resulting in lack of Fas expression on cell membrane. It has been previously shown that cloned Fas-negative but not Fas-positive human cholangiocarcinoma cells are resistant to anti-Fas-mediated apoptosis and develop tumors in nude mice. In addition, interferon gamma induces Fas expression in Fas negative cholangiocarcinoma cells and makes them susceptible to apoptosis. Therefore, it becomes important to characterize immunophenotypic expression of p53 and Fas in normal and neoplastic human tissues of the biliary tract to further understand the pathogenesis of the disease. To date, human studies to characterize differences in immunophenotypic expression of the Fas protein between intrahepatic and extrahepatic biliary duct cancers and in their precursor lesions have not been performed. OBJECTIVE: To report the immunophenotypic expression of p53 and Fas expression in various stages in the development of bile duct cancers (intrahepatic and extrahepatic tumor location) and their association with tumor differentiation. DESIGN: Thirty bile duct cancer samples (13 intrahepatic and 17 extrahepatic) from 18 men and 12 women who ranged in age from 44 to 77 years (mean age, 65.6 years) were retrieved from the surgical pathology files. Hematoxylin-eosin-stained slides were evaluated for the type and grade of tumor and dysplastic changes in the biliary tract epithelium. Additional slides were immunohistochemically stained with p53 and anti-Fas mouse monoclonal antibody. The pattern of Fas distribution and percentage of cells positive for p53 and Fas expression were determined. RESULTS: The percentage of Fas-expressing cells is significantly (P = .01) more frequently noted in extrahepatic tumors compared with intrahepatic tumors. Furthermore, Fas expression decreased from dysplastic epithelium to cholangiocarcinoma (P = .01), and this decreasing trend continued from well to poorly differentiated tumors. Nuclear p53 expression was not identified in normal and dysplastic epithelium but was noted in 30% of carcinomas (P = .02). CONCLUSION: Fas expression is an early event in pathogenesis of bile duct cancers. Immunophenotypic expression of Fas is associated with well to moderately differentiated tumors but not with poor tumor differentiation. PMID- 15794671 TI - Renal neoplasms in younger adults: analysis of 112 tumors from a single institution according to the new 2004 World Health Organization classification and 2002 American Joint Committee on Cancer Staging System. AB - CONTEXT: Adult renal neoplasms have a predilection for older patients and are clinically and morphologically distinct from renal neoplasms found in pediatric age groups. Relatively rare tumors occur in younger adults (18-45 years of age). Whether these renal tumors are morphologically and clinically distinct from those of older adults has been the subject of controversy. Recent modification of the World Health Organization histologic classification and the American Joint Committee on Cancer staging system of adult renal tumors further highlighted the need for case analysis in this age group. OBJECTIVE: To analyze renal tumors in younger adults based on a large surgical series from a single institution. DESIGN: Of 780 renal mass nephrectomy (partial, total, or radical) specimens that were available for evaluation and had been obtained between 1986 and 2004 at Loyola University Medical Center, 112 specimens were from patients between 18 and 45 years of age. The tumors were reevaluated according to the 2004 World Health Organization classification and the 2002 American Joint Committee on Cancer staging system. RESULTS: The likelihood of clear cell renal cell carcinoma was significantly reduced from 65% in older adults to 53% in younger adults (18-45 years, P = .04). The reduction trend was more significant when comparing an even younger age group. The majority (64%) of clear cell renal cell carcinoma in younger adults was low stage, T1a. Seventeen percent of these tumors had multilocular cystic features involving more than 50% of the tumor volume (55% 85%). The number of oncocytomas was also significantly lower in younger adults than in older adults (2% vs 11%, P < .001), and this presumably age-related benign neoplasm was not identified in patients younger than 40 years in this study. In contrast, the miscellaneous tumor category showed a remarkable increase, from 4% in older adults to 26% in younger adults (P < .001). The youngest patient group (18-35 years) had a higher incidence of miscellaneous tumors, 37%. Younger female adults tended to have more benign miscellaneous neoplasms than did their male counterparts (64% vs 36%, P < .001). Clear cell and chromophobe renal cell carcinoma occurred more frequently in younger male adults than in female adults (2:1 and 8:1, respectively). CONCLUSIONS: Renal neoplasms are more heterogeneous in younger adults and have a different distribution pattern compared with that in older adults. Malignant and benign renal neoplasms tend to have a contrasting sex distribution in younger adults. PMID- 15794672 TI - A procedure for rapid issue of red cells for emergency use. AB - CONTEXT: A College of American Pathologists Q-Probe revealed that the median turnaround times for emergency requests for red blood cells from the operating room were 30 minutes to release of cells from the blood bank and 34 minutes to delivery to the operating room. These times may not be adequate to permit the red cells to provide sufficiently rapid delivery of oxygen in massively bleeding patients. OBJECTIVE: To improve the time from emergency request for red cells to delivery to the operating room. DESIGN: A new emergency issue program was implemented for only the operating rooms; emergency issue to all other hospital locations remained unchanged. Six units of group O Rh-negative red blood cells (RBCs) are maintained in the blood bank in a separate basket with transfusion forms containing the unit numbers and expiration dates and a bag with one blood tubing segment from each unit. The times to issue and to delivery to the operating room suite were compared with time to issue of 2 group O Rh-negative RBCs for other hospital locations using the older system during the same time period and with the time to issue of 2 units to all other hospital locations during the preceding 2 years. SETTING: A university hospital. MAIN OUTCOME MEASURES: Time between emergency request for red cells and delivery to the operating room. RESULTS: The time between blood bank notification and arrival in the operating room of the 6 units of RBCs was significantly shorter than the time required to just issue (not including delivery time) 2 units of RBCs to other hospital locations. With the new procedure, 82% of units issued reached the operating room within 2 minutes of request, 91% arrived within 3 minutes, and 100% arrived within 4 minutes. These percentages are significantly higher than those for only issue of blood (without delivery) using the older issuing procedure for all hospital locations during the previous 2 years (37%, 49%, and 66%, respectively; P = .007, .009, and .02, respectively) and for other locations during the same 7-month period (29%, 46%, and 73%, respectively; P = .004, .01, and .09, respectively). Time (mean [95% confidence interval]) from blood bank notification to delivery of RBCs to the operating room suite (2.1 [1.6-2.6] minutes, of which approximately 50-60 seconds is attributable to delivery time) was less than issue times (not including delivery times) using the older issuing procedure for other hospital locations during the same period (4.1 [3.1-5.0] minutes; P = .007). CONCLUSIONS: An emergency issue procedure can be used to issue several units of RBCs within 1 minute and have them delivered to the operating room within 2 minutes while maintaining sufficient controls and providing required information to satisfy patient and blood bank requirements. PMID- 15794673 TI - Comparison of relative value of bone marrow aspirates and bone marrow trephine biopsies in the diagnosis of solid tumor metastasis and Hodgkin lymphoma: institutional experience and literature review. AB - CONTEXT: Bone marrow aspirates as well as bone marrow trephine biopsies are frequently performed to assess whether there is marrow involvement by a malignancy. Numerous reports differ in the relative value of these 2 procedures and fail to provide concise guidelines that can help choose the appropriate technique in this clinical situation. OBJECTIVE: To compare the relative value of aspirates and trephine biopsies in the diagnosis of solid tumor metastasis and Hodgkin lymphoma. In addition, we correlate our findings with those of the literature to provide a concise practice guideline. DESIGN: Sixty-six cases showing bone marrow involvement by solid tumor and Hodgkin lymphoma in bone marrow aspirates, bone marrow trephine biopsies, or both were included in the study. The diagnosis and findings made on aspirates were compared with those made on trephine biopsies in each case. RESULTS: In those cases where both aspirate and trephine biopsy were available for evaluation, there was a 22% positive correlation in the findings on aspirates and trephine biopsies. The correlation between aspirates and trephine biopsies was highest in cases of small cell carcinoma of the lung (3/11, or 36.3%) followed by breast carcinoma (7/20, or 35%), prostate carcinoma (1/9, or 11.1%), and Hodgkin lymphoma (1/20, or 5%). Two of 5 cases from the miscellaneous category demonstrated simultaneous involvement of aspirate and trephine biopsy by a gastric carcinoma as well as an adrenal gland carcinoma. CONCLUSIONS: Bone marrow aspirate and bone marrow trephine biopsy should both be performed in patients with proven or suspected malignancies where staging may affect management. However, bone marrow aspirate has only a minimal role, if any, in detecting bone marrow involvement by Hodgkin lymphoma. In cases of breast carcinoma, small cell carcinoma of lung, and prostate carcinoma, aspirate evaluation may confirm trephine biopsy results or, more rarely, provide the sole confirmation of the malignancy. PMID- 15794674 TI - Histomorphologic assessment of formalin substitute fixatives for diagnostic surgical pathology. AB - CONTEXT: In 1987, the Formaldehyde Standard became law in the United States, alerting laboratory workers to the potential carcinogenicity of formaldehyde. As a result, a variety of proprietary fixatives were developed for use in surgical pathology. OBJECTIVE: To assess histomorphology with different formalin substitute fixatives. DESIGN: Four experienced board-certified surgical pathologists examined 7 specimens (hepatocellular carcinoma, ovarian sex cord/stromal tumor, myxoid liposarcoma, uterine endometrioid adenocarcinoma, splenic follicular hyperplasia, infiltrating mammary carcinoma, and cecal signet ring carcinoma) fixed with formalin and 5 proprietary fixatives advertised as formalin replacements. In a blind study, the pathologists rated cellular outlines, cytoplasmic detail, nuclear detail, erythrocyte integrity, lymphocyte integrity, overall morphology, and overall staining in each case. RESULTS: Formalin received the highest overall morphology and staining scores, followed by Glyo-Fixx, STF-Streck, Omnifix, Histochoice, and Histofix. Formalin also received the highest scores in cellular outline and erythrocyte integrity. Individually, some fixatives performed better in different areas than others. Glyo-Fixx performed as well as formalin for overall morphology and provided highest nuclear detail and lymphocyte appearance scores. Omnifix II gave best results for cytoplasmic detail. CONCLUSION: In this blind study, formalin fixation provided the highest histomorphologic quality for tissue stained with hematoxylin-eosin and examined for diagnostic surgical pathology. Should the use of formalin be discontinued, pathologists will have to familiarize themselves with a different set of microscopic details associated with the replacement fixatives. PMID- 15794675 TI - Three-dimensional virtual microscopy of colorectal biopsies. AB - Conventional optical microscopy of specimens from colorectal biopsies commonly produces diagnostic errors due to incomplete sampling or poor orientation. Obtaining additional sections or re-embedding may help avoid these errors, but can prolong turnaround time. We describe new technology, which incorporates exhaustive sectioning, 3-dimensional reconstruction, and virtual microscopy, that may eliminate these problems by enabling pathologists to rapidly examine entire specimens and convert poorly oriented mucosa to well-oriented mucosa. PMID- 15794676 TI - Cardiac aspergillosis in patients with acquired immunodeficiency syndrome: a case report and review of the literature. AB - Cardiac aspergillosis is uncommon in patients with acquired immunodeficiency syndrome (AIDS) in the absence of open heart surgery. We report a unique case of a 62-year-old man with AIDS who developed Aspergillus pancarditis with Aspergillus vegetations on mitral valve without evidence of pulmonary aspergillosis. There was extensive embolization to the brain and multiple foci of Aspergillus infection in kidneys and adrenal glands. There are only 10 documented cases of cardiac aspergillosis in the literature (1966-2003) in severely immunocompromised AIDS patients with CD4 T-lymphocyte counts ranging from 10 to 121 cells/muL. The cardiac aspergillosis could result from invasive pulmonary aspergillosis, either by hematogenous dissemination or by direct invasion, and skin Aspergillus infection can be carried through the bloodstream to the right heart in intravenous drug abusers. Most of the reported cases of cardiac aspergillosis were diagnosed at autopsy. Mortality among AIDS patients with cardiac aspergillosis is 100%, despite appropriate therapy. PMID- 15794677 TI - Intravascular papillary endothelial hyperplasia of renal vein: report of 2 cases. AB - Intravascular papillary endothelial hyperplasia is a benign intravascular process thought to arise from an organizing thrombus. The lesion may present clinically as an abnormal mass and, depending on the location, may be confused with benign or malignant neoplasms. It has been described in a variety of locations. Involvement of the renal vein by papillary endothelial hyperplasia is extremely rare, with only 4 cases reported in the literature. We describe 2 additional cases. In both cases, the radiologic examination revealed a well-circumscribed mass in the hilar region of the kidney, which was considered to be a renal neoplasm. Nephrectomy specimen in each case revealed characteristic features of intravascular papillary endothelial hyperplasia. It is suggested that intravascular papillary endothelial hyperplasia should be included in the differential diagnosis of a hilar renal mass. PMID- 15794678 TI - Fibrous hamartoma of infancy: a case report with associated cytogenetic findings. AB - A 2-month-old male infant presented with a subcutaneous mass on the left middle finger; the mass had been present since birth. This was treated with local excision, and there has been no recurrence. Histology revealed the typical features of a fibrous hamartoma. Cytogenetic studies revealed a reciprocal translocation, t(2;3)(q31;q21), as the sole abnormality. To our knowledge, this is the first report of the cytogenetic findings in fibrous hamartoma, and it suggests that this lesion represents a benign neoplasm. PMID- 15794679 TI - Gallbladder paraganglioma: a case report with review of the literature. AB - We report a case of gallbladder paraganglioma that was discovered during nonrelated surgery. Retrospective study disclosed a family history of pheochromocytoma. The occurrence of gallbladder paraganglioma in the presence of family history of endocrine neoplasia supports that gallbladder paraganglioma may indeed occur as a part of the multiple endocrine neoplasm syndrome. Gallbladder paraganglioma is a rare tumor, and so far to our knowledge only 6 cases have been reported in the literature. Three cases were discovered incidentally during cholecystectomy for cholelithiasis, 2 presented with right upper quadrant pain, and 1 manifested with gastrointestinal bleeding. We herein review all reported cases of paraganglioma of gallbladder and biliary system. PMID- 15794680 TI - An unusual case of peripheral T-cell lymphoma with CD56 positivity and angiocentric, angiodestructive morphology arising in the ileum. AB - Natural killer cell and cytotoxic T-cell lymphomas are frequently difficult to distinguish because they share many common features, and yet it is important to make an accurate diagnosis because their prognoses differ. We report an unusual case of a white man with a CD56-positive T-cell lymphoma in the ileum. The histologic pattern was characterized by angioinvasion and angiodestruction. Immunohistochemical staining showed positive reactions to CD3, CD8, CD43, CD45RO, CD56, and T-cell intracellular antigen-1, but negative reactions to CD4, CD5, CD20, CD23, and CD57. Epstein-Barr virus (EBV) was not detected by EBV-latent membrane protein staining and EBV polymerase chain reaction technique. The T-cell receptor gamma chain gene was rearranged. According to the World Health Organization classification, the absence of EBV excludes the diagnosis of extranodal natural killer/T-cell lymphoma, nasal type. However, the association of EBV with this lymphoma in white patients is not clear. Therefore, absence of EBV alone does not necessarily exclude nasal-type natural killer/T-cell lymphoma, particularly because the histologic pattern in this case is highly characteristic of this tumor. PMID- 15794681 TI - Small cell carcinoma of the ovary with hypercalcemia and ectopic parathyroid hormone production. AB - Small cell carcinoma of the ovary is a rare malignant tumor of the ovary. It is the most common undifferentiated ovarian carcinoma in young women. Approximately two thirds of patients with ovarian small cell carcinoma have hypercalcemia. The mechanism of development of hypercalcemia is unclear, although parathyroid hormone-related protein has been found in some of the cases. Parathormone expression in tumor cells, rarely reported, was seen in this case, suggesting that ectopic parathyroid hormone production by the tumor cells may be the cause of hypercalcemia. PMID- 15794682 TI - Pathologic quiz case: an orbital mass in a 45-year-old woman. Alveolar soft part sarcoma. PMID- 15794684 TI - Pathologic quiz case: a 33-year-old woman with a large temporal mass. Chondroblastoma of the skull. PMID- 15794683 TI - Pathologic quiz case: a lymph node lesion in a 58-year-old woman with invasive high-grade urothelial carcinoma. Lymph node inclusion of serous borderline tumor. PMID- 15794685 TI - Pathologic quiz case: a 64-year-old man with an adrenal mass. Myxoid adrenal cortical neoplasm. PMID- 15794686 TI - Nebulous corpora amylacea. PMID- 15794687 TI - Schistosoma mansoni in colon and liver. PMID- 15794688 TI - Smiley face. PMID- 15794690 TI - Pathologic quiz case: a 58-year-old woman with a slowly enlarging parotid mass. Pleomorphic adenoma with abundant tyrosine crystalloids. PMID- 15794691 TI - Pathologic quiz case: a 53-year-old woman with bilateral ovarian tumors. Double tumors, primary mucinous bilateral ovarian carcinoma (G2) with metastasis to myometrium and primary intestinal carcinoma (G2) infiltrating the serosal surface with metastasis to one peri-intestinal lymph gland. PMID- 15794692 TI - Pathologic quiz case: a 13-year-old adolescent girl with an abdominal mass 2 months after radiation for Hodgkin lymphoma. Epithelioid angiosarcoma. PMID- 15794693 TI - Pathologic quiz case: a 16-year-old adolescent girl with a trace level of hemoglobin s detected by high-performance liquid chromatography. Almost complete suppression of hemoglobin S in a sickle cell disease patient undergoing long-term transfusion therapy. PMID- 15794694 TI - Pathologic quiz case: a woman with chronic pelvic pain. Endocervicosis. PMID- 15794696 TI - Gastric toxoplasmosis as the presentation of acquired immunodeficiency syndrome. AB - We report a case of a 39-year-old West African man with unknown human immunodeficiency virus status diagnosed with gastric toxoplasmosis as the presenting manifestation of acquired immunodeficiency syndrome. Toxoplasma gondii is common in severely immunosuppressed patients and most frequently involves the central nervous system, followed by the eye, myocardium and skeletal muscle, lungs, bone marrow, and peripheral blood. For unclear reasons, gastrointestinal involvement is exceedingly rare and occurs in the context of severe immunosuppression and disseminated disease. To our knowledge, this is the first report in the English literature of a patient with isolated, manifest gastric toxoplasmosis without evidence of concomitant cerebral or extracerebral involvement. It is important for both the clinician and the pathologist to maintain a high index of suspicion for toxoplasmosis in immunosuppressed patients presenting with nonspecific symptoms of gastritis and radiologic and endoscopic presence of thickened gastric folds with or without ulceration. PMID- 15794695 TI - Pathologic quiz case: twin neonates with thrombocytopenia. May-Hegglin anomaly. PMID- 15794697 TI - Periurethral carcinosarcoma: a report of 2 cases with a review of the literature. AB - Carcinosarcomas are rare, aggressive tumors that can arise in various sites of the body, especially the genitourinary system. In this system, the most common location for the tumor is the uterus in the female and the urinary bladder in the male. To our knowledge, only 2 cases with a periurethral location for this tumor have been previously described in the English literature. We describe 2 additional cases in this location, one in either sex, with a review of the relevant literature. PMID- 15794698 TI - Basaloid squamous carcinoma of the hypopharynx with an extensive spindle cell component exhibiting a pedunculated polypoid mass. AB - We report a case of basaloid squamous carcinoma of the hypopharynx with an extensive spindle cell component in an 82-year-old man. The tumor (4.0 x 2.5 x 2.2 cm) was a pedunculated polypoid mass that was attached to the left pyriform sinus. Histologically, most (70%) of the tumor was composed of malignant spindle cell proliferation, and the rest (30%) was basaloid squamous carcinoma. Some of the spindle cells were positive for cytokeratins. The preoperative histologic diagnosis was difficult because the endoscopic biopsy specimens showed only spindle cell proliferation. This is an unusual presentation of basaloid squamous carcinoma of the head and neck. PMID- 15794700 TI - The peril of inadequate evidence. PMID- 15794699 TI - Pseudofungi in pericolic lymph nodes. AB - We report a case of extensive pseudofungi in the pericolic lymph nodes. A 45-year old man presented with lower abdominal discomfort, and large solid and cystic masses were discovered in the pelvic cavity by computed tomography. Debulking of the masses (by means of a Hartmann operation) was performed under the clinical impression of a pelvic sarcoma, and histologically the masses demonstrated features of a malignant gastrointestinal stromal tumor. All pericolic lymph nodes demonstrated multiple septate hyphae-like structures with pigmentation, particularly in the subcapsular sinuses. These structures showed positive staining with periodic acid-Schiff, but Gomori methenamine silver staining was negative. The structures appeared to be composed of iron, phosphorus, and calcium by Perls iron and von Kossa stains and revealed deposition of electron-dense granules on electron microscopic examination. Energy-dispersive radiographic micro-analysis showed the structures to contain phosphorus, sulfur, potassium, calcium, and iron. Pseudofungi can be misinterpreted as true fungi with septate hyphae; therefore, a careful morphologic examination with appropriate special stains is mandatory. PMID- 15794701 TI - Clinical examination variables discriminate among treatment-based classification groups: a study of construct validity in patients with acute low back pain. AB - BACKGROUND AND PURPOSE: Treatment-based classification (TBC) provides matched interventions for patients with acute low back pain (LBP) through key history and clinical findings. This study investigated the discriminant validity of TBC by determining whether commonly used clinical examination variables discriminated among TBC groups. SUBJECTS: The mean age of the 131 participants was 37.7 years (SD=10.1), 66 participants (50.4%) were female, mean duration of LBP was 16.5 days (SD=16.1), and 60 participants (45.8%) had a prior history of LBP. Fifty-one study participants (38.9%) were classified for specific exercise, 42 (32.1%) for mobilization, 28 (21.4%) for immobilization, and 10 (7.6%) for traction. METHODS: One-way analyses of variance and Kruskal-Wallis tests were used to investigate differences in clinical variables by TBC group. Then, discriminant function analysis (DFA) predicted TBC group membership. RESULTS: The TBC groups differed on present pain intensity, duration of LBP, and history of LBP. Present pain intensity, duration of LBP, total lumbar flexion, presence of leg pain, and history of LBP produced 2 statistically significant discriminant functions that predicted TBC group membership. These functions correctly classified (cross validation value in parentheses) 65% (65%) for specific exercise, 45% (40%) for mobilization, and 32% (32%) for immobilization. DISCUSSION AND CONCLUSION: This study provided evidence supporting the discriminant validity of TBC. Additional diagnostic information related to TBC groups was generated. PMID- 15794702 TI - Factors influencing results of functional capacity evaluations in workers' compensation claimants with low back pain. AB - BACKGROUND AND PURPOSE: Physical and psychosocial factors are hypothesized to influence performance-based assessment. The purpose of this study was to evaluate the association between performance on the Isernhagen Work System Functional Capacity Evaluation (IWS-FCE) and various clinical and psychosocial factors. SUBJECTS: The sample consisted of 170 workers' compensation claimants who were undergoing functional capacity evaluations (FCEs) for low back injuries. METHODS: In this cross-sectional study, claimants completed a battery of work-related measures, including the IWS-FCE, the Pain Disability Index (PDI), a workplace organizational policies and practices scale, and a recovery expectations questionnaire. Functional capacity evaluation performance indicators were the number of tasks in which subjects did not meet work demands and weight lifted on the floor-to-waist lift. Analysis included multivariable regression. RESULTS: Only the PDI, pain intensity, age, and sex independently contributed to floor-to waist lift performance. The PDI, pain intensity, and duration of injury contributed to the number of failed tasks. DISCUSSION AND CONCLUSION: The results indicate that performance on FCEs is influenced by physical factors, perceptions of disability, and pain intensity. However, perceptions of workplace organizational policies and procedures were not associated with FCE results for workers' compensation claimants with chronic back pain disability. Functional capacity evaluations should be considered behavioral tests influenced by multiple factors, including physical ability, beliefs, and perceptions. PMID- 15794703 TI - Continuous-scale physical functional performance test: validity, reliability, and sensitivity of data for the short version. AB - BACKGROUND AND PURPOSE: The Continuous-Scale Physical Functional Performance Test (CS-PFP) can be used to obtain valid, reliable, and sensitive measurements of physical functional capacity. This test requires a fixed laboratory space and approximately 1 hour to administer. This study was carried out in 4 steps, or substudies, to develop and validate a short, community-based version (PFP-10) that requires less space and equipment than the CS-PFP. SUBJECTS AND METHODS: Retrospective data (n=228) and prospective data (n=91) on men and women performing the CS-PFP or the PFP-10 are reported. A 12-week exercise program was used to examine sensitivity to change. Data analyses were done using paired t test, Pearson correlation, intraclass correlation coefficient (ICC), and delta index (DI) procedures. RESULTS: The PFP-10 total score and 4 of the 5 domain scores were statistically similar (within 3%) to those of the CS-PFP. The PFP-10 upper-body strength domain score was 17% lower, but was highly correlated (ICC=.97). Community and established laboratory PFP-10 scores were similar (ICC=.85-.97). The PFP-10 also is sensitive to change (DI=.21-.54). DISCUSSION AND CONCLUSION: The PFP-10 yields valid, reliable, and sensitive measurements and can be confidently substituted for the CS-PFP. PMID- 15794704 TI - Classification, intervention, and outcomes for a person with lumbar rotation with flexion syndrome. AB - BACKGROUND AND PURPOSE: The purpose of this case report is to describe the classification, intervention, and outcomes for a patient with lumbar rotation with flexion syndrome. CASE DESCRIPTION: The patient was a 22-year-old man with a medical diagnosis of low back strain. Impairments in lumbar flexion and right rotation and lateral bending were identified. Daily activities and positions associated with these actions were associated with increased low back pain (LBP). Instruction focused on modifying lumbar rotation and flexion movements and alignments in daily activities. Exercises to address the direction-specific impairments were prescribed. OUTCOMES: The patient participated in 4 visits and completed a questionnaire 1 year after intervention. The patient reported a decrease in symptoms, disability, and frequency of recurrences. DISCUSSION: Repetition of specific strategies (alignment and movement) during activities may result in specific impairments that contribute to LBP. Modification of the strategies and exercises to change contributing factors are proposed to help alleviate symptoms, disability, and recurrences. PMID- 15794705 TI - Ultrasound therapy in rectus sheath hematoma. AB - BACKGROUND AND PURPOSE: Ultrasound (US) is one of the most common modalities used in intervention for musculoskeletal disorders, although its effectiveness is debated. The purpose of this case report is to describe the intervention, including the use of US, in the management of a large rectus sheath hematoma (RSH) in a patient receiving anticoagulant therapy. CASE DESCRIPTION: The patient was a 62-year-old woman with RSH who was receiving oral anticoagulant therapy and had a history of bouts of coughing. Computed tomographic scans verified the diagnosis of RSH. The report describes the patient examination, management, intervention, and outcomes. OUTCOMES: The intervention, including the use of US therapy, may have enabled a rapid resolution of the hematoma. DISCUSSION: This case report illustrates how US may be a useful modality for complementary management of RSH, helping the reabsorption of the hematoma. PMID- 15794706 TI - Recruitment patterns in human skeletal muscle during electrical stimulation. AB - Electromyostimulation (EMS) incorporates the use of electrical current to activate skeletal muscle and facilitate contraction. It is commonly used in clinical settings to mimic voluntary contractions and enhance the rehabilitation of human skeletal muscles. Although the beneficial effects of EMS are widely accepted, discrepancies concerning the specific responses to EMS versus voluntary actions exist. The unique effects of EMS have been attributed to several mechanisms, most notably a reversal of the recruitment pattern typically associated with voluntary muscle activation. This perspective outlines the authors' contention that electrical stimulation recruits motor units in a nonselective, spatially fixed, and temporally synchronous pattern. Furthermore, it synthesizes the evidence that supports the contention that this recruitment pattern contributes to increased muscle fatigue when compared with voluntary actions. The authors believe the majority of evidence suggests that EMS-induced motor unit recruitment is nonselective and that muscle fibers are recruited without obvious sequencing related to fiber types. PMID- 15794707 TI - Diaphragmatic breathing training: further investigation needed. PMID- 15794708 TI - Diaphragmatic breathing training: further investigation needed. PMID- 15794709 TI - What to do with TNF failures. AB - The introduction of TNF inhibitors into the clinic for the treatment of patients with rheumatoid arthritis and other systemic inflammatory conditions has revolutionised the approach to these diseases. Despite the substantial efficacy of these agents, a subset of patients either does not respond or has a suboptimal clinical response. Defining what constitutes 'failure' of this type of therapy is important in both clinical research and practice. For patients who fail to respond to TNF inhibitors, a number of promising avenues targeting other aspects of the immune system are under study, and may be brought to the clinic in the near future. PMID- 15794710 TI - Gastrointestinal adverse effects of non-steroidal anti-inflammatory drugs. AB - NSAIDs are used extensively worldwide at a cost of billions of dollars annu-ally. Adverse side effects, especially in the gastrointestinal (GI) tract, are uncommon but cause a substantial burden of illness because of the volume of use. Important upper GI complications include dyspepsia, gastric erosions and peptic ulcers and complications such as bleeding, perforation or gastric outlet obstruction. Dyspeptic symptoms may occur without correlation to endoscopic findings. Topical injury and COX-1 inhibition resulting in gastric prostaglandin suppression are two commonly postulated mechanisms of gastroduodenal damage. Advanced age, previous peptic ulcers or ulcer complications, concomitant use of glucocorticoids or anticoagulants, and high-dose or prolonged NSAID administration are known risk factors. Prevention of adverse GI events involves use of safer NSAIDs including COX-2 inhibitors, and co-prescription of gastroprotective agents. NSAID-induced ulcers heal with proton pump inhibitors or misoprostol. The role of Helicobacter pylori eradication in NSAID ulcer prophylaxis and management is controversial. Choice of NSAIDs and gastroprotective agents should be guided by risk/benefit and cost-effectiveness assessment. PMID- 15794711 TI - Benefits and risks assessment of simvastatin in familial hypercholesterolaemia. AB - Familial hypercholesterolaemia (FH) is a frequent inherited monogenic disorder, associated with premature coronary artery disease. Life expectancy of FH patients is reduced by 15 - 30 years unless they are adequately treated with lipid lowering therapy. Patients with this disorder need long-term drug therapy and the selection of treatment should be strongly based on its long-term safety and tolerability. The introduction of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors has changed the treatment of FH. Simvastatin 40 - 80 mg/day effectively reduces serum low-density lipoprotein cholesterol levels, and also reduces triglycerides with a modest rise in high-density lipoprotein cholesterol levels. Other potentially important effects, such as improvement of endothelial function, reduction of LDL oxidation and vascular inflammation, have been associated with simvastatin therapy in FH. In addition, simvastatin has been shown to abolish the progression, and even facilitate the regression of existing human atherosclerotic lesions. The safety and tolerability of simvastatin is clearly highlighted by the low rate of therapy discontinuation observed in several population-based clinical trials. Asymptomatic elevations in liver transaminase levels and myopathy are uncommon. The efficacy and tolerability of simvastatin at doses up to 80 mg/day are well-established, as well as its cost effectiveness in the management of FH patients. PMID- 15794712 TI - The development and application of imatinib. AB - The hallmark characteristics of cancer include an unrestrained proliferation involving activation of growth signals, loss of negative regulation and dysfunctional apoptotic pathways. Targeting abnormal cell signalling pathways should provide a more selective approach to cancer treatment than conventional cytotoxic chemotherapy. Tyrosine kinases play an essential role in the signalling pathways involved in the control of cellular proliferation and growth. Imatinib is a small-molecule tyrosine kinase inhibitor of the ABL fusion gene, platelet derived growth factor receptors (PDGFR) and KIT. This agent has demonstrated considerable activity in chronic myeloid leukaemia (CML) by inhibiting the BCR ABL fusion protein and gastrointestinal stromal tumours (GISTs), which are predominantly driven by activating mutations in KIT. A number of other rare conditions are also responsive, for example, dermatofibrosarcoma protuberans, which is driven by a chromosomal translocation involving PDGF-B and Col1A1, resulting in overexpression of PDGF-B, and hypereosinophillic syndrome, which can be caused by activating PDGFR mutations. The pivotal registration study for newly diagnosed CML was a large randomised trial comparing 400 mg/day of imatinib to a combination of IFN-alpha and cytarabine, which demonstrated a significantly higher complete haematological and cytogenetic response rate in the imatinib arm. In the case of GIST a randomised study in patients with inoperable or metastatic disease explored doses of 400 - 600mg and reported a response rate of > 50% in each arm plus disease stabilisation and an improvement in performance status. Large randomised trials have subsequently been performed, comparing 400 with 800mg/day. The first to report indicates that the larger dose is associated with improved progression-free survival, although it is not yet known whether or not this will translate into a difference in overall survival. The most common KIT mutation involves exon 11 and is associated with a statistically significant better response and prognosis compared with other mutations or no detectable mutations. Mutational analysis is likely to become increasingly important in the selection of patients for neoadjuvant and adjuvant treatment and in helping to understand the nature of acquired resistance. PMID- 15794713 TI - Chemotherapy-induced oral mucositis: new approaches to prevention and management. AB - Oral mucositis is a common and significant toxicity of cancer chemotherapy. It is under-reported and not well treated, particularly in patients that receive high dose therapy with an autologous or allogenic stem cell transplant. Two recently published retrospective analyses of patient complaints following stem cell transplantation have identified oral mucositis as the worst toxicity reported by patients, and what is more important is that patients indicated that oncology healthcare team members do a poor job of managing and providing methods of symptom relief. Twenty percent of patients surveyed indicated they received no symptom relief at all. PMID- 15794715 TI - Toxicity of the topoisomerase II inhibitors. AB - Topoisomerase II inhibitors represent a broad class of antineoplastic agents with a wide spectrum of activity against malignancies. Topoisomerase II inhibitors include the anthracyclines, mitoxantrone and epipodophyllotoxins. Short-term toxicity includes myelosuppression and gastrointestinal toxicity. Long-term survivors are at risk of cardiac toxicity and secondary leukaemia. This article discusses these toxicities in detail, including administration of these agents to patients with hepatic and/or renal insufficiency, and the need for dose adjustments in selected patient populations. PMID- 15794714 TI - Adverse effects of antiretroviral therapy for HIV infection: a review of selected topics. AB - In the current era of HIV treatment, the toxicity profiles of antiretroviral drugs have increasingly emerged as a basis for selecting initial antiretroviral regimens as well as a reason for switching therapy in treatment-experienced patients. In this respect, an intensive research effort involving clinical research as well as basic science research over the past six years, has focused on the cluster of metabolic and body composition abnormalities that have come to be termed the 'lipodystrophy syndrome'. These data have now provided a clear and clinically relevant understanding of the individual profiles of drugs within the nucleoside analogue reverse transcriptase inhibitor , HIV protease inhibitor and non-nucleoside analogue reverse transcriptase inhibitor drug classes, and have provided a rational basis for assessing and monitoring these adverse effects in clinical practice. In this review, current and emerging drug toxicities are considered with an emphasis on lipodystrophy complications. PMID- 15794716 TI - The safety and efficacy of ciclopirox olamine for the treatment of seborrheic dermatitis. AB - Ciclopirox olamine is a synthetic hydroxypiridone derived, broad spectrum, antifungal agent which has been used effectively to treat seborrheic dermatitis. Seborrheic dermatitis is a chronic dermatosis, more common in men than women, which usually occurs in sites dense with sebaceous glands in the form of mild inflammatory desquamate erythema. Treatment modalities for seborrheic dermatitis include keratolytic agents, corticosteroids and antifungal agents. Due to its antimycotic and anti-inflammatory activities, ciclopirox olamine is established as an effective treatment for this condition. PMID- 15794717 TI - The safety and tolerability of botulinum toxins for the treatment of cervical dystonia. AB - The use of botulinum toxin to treat cervical dystonia (CD) has dramatically improved the quality of life of patients with this disabling, often painful disease. Two forms of type A toxin (BOTOX and Dysport) and one form of type B toxin (MyoBloc) are available in some parts of the world to treat patients with CD. The literature supports the efficacy of each in reducing the pain and movement of cervical dystonia. The dosing and side effects vary between the toxins. The potential availability of several forms of toxin will allow physicians to offer further treatment options to patients with CD. However, it is incumbent on the treating physicians to have a working knowledge of the different serotypes, different doses used of each formulation of each serotype, the side effect profile of each product and the potential for anti-body formation for each form of toxin. PMID- 15794718 TI - The risk/benefit of inhaled corticosteroids in chronic obstructive pulmonary disease. AB - Although inhaled corticosteroids have a well defined role in asthma therapy, their use remains controversial in nonasthmatic, smoking-related chronic obstructive pulmonary disease (COPD). Some studies have shown an effect of inhaled corticosteroids on airway inflammation in COPD, but the clinical relevance of these results is unknown. Data from five long-term, large studies, provide evidence that prolonged treatment with inhaled corticosteroids does not modify the rate of decline of forced expiratory volume in one second (FEV1) in patients with COPD and no reversibility to short-acting beta(2)-agonists. FEV1 was slightly improved over the first six months of treatment and lower reactivity in response to methacholine challenge has been observed. Improvement of respiratory symptoms and health status were also reported. A reduction of exacerbations rate was observed in two studies. No survival benefit was demonstrated. Two recent reports suggest that long term use of inhaled corticosteroids in COPD patients improves quality-adjusted life expectancy and is cost-effective. Combination therapy with inhaled corticosteroids and long-acting beta(2)-agonists have proven benefit in four long term large studies compared to placebo for FEV1, exacerbation rate, symptoms and health status. However, only two studies found that combination therapy was more effective than long-acting beta(2)-agonists alone for symptoms and health status improvement. The long term safety of inhaled corticosteroids is not known in COPD patients but topical adverse effects, and systemic effects such as a decrease of bone density of lumbar spine and femur and cutaneous adverse effects, have been reported after three years of treatment. However, three recent observational studies found a slight increase in the risk of fractures (hip, upper extremities and vertebral) in association with high doses of inhaled corticotherapy. PMID- 15794719 TI - The safety of antidepressant use in pregnancy. AB - Depression during pregnancy affects an estimated 10 - 20% of women, some of whom will require treatment with antidepressants. It is of great importance that the safety of this particular class of drugs is reviewed, to ensure the optimal treatment of the mother while protecting her unborn child. In this review, current safety data on all available antidepressants are discussed in detail, including the pharmacokinetics of the maternal-fetal unit and the epidemiological studies that have been published to date. The classes of antidepressants discussed include: tricyclics, selective serotonin re-uptake inhibitors and other antidepressants. After reviewing these studies, it is evident that these drugs appear to be relatively safe to take during pregnancy. This evidence-based information will be helpful to women and their healthcare providers, when the decision of whether or not to treat with anti-depressants during pregnancy must be made. PMID- 15794720 TI - Drug-induced aseptic meningitis. AB - Drug-induced aseptic meningitis (DIAM) is an important entity. This article reviews the literature on this rare idiosyncratic event which may occur after local or systemic drug administration. The data on this adverse reaction is predominantly collated from anecdotal case reports and case series. PMID- 15794721 TI - Fluoroquinolones and tendon disorders. AB - Fluoroquinolones are the most potent oral antibiotics in clinical use today. Increasingly, these drugs are being prescribed for relatively benign infections and for new categories of patients, including paediatric patients. As their use becomes more frequent, so will the adverse events. This review focuses on a rare but debilitating adverse reaction, the fluoroquinolone-associated tendinopathy. Despite many published case reports and approximately 3500 cases reported to the World Health Organization Collaborating Centre for Drug Monitoring, little is known about the mechanisms behind this fluoroquinolone-specific toxicity. Data on chemical properties, mode of action, pharmacokinetic features, clinical presentation and risk factors in relation to tendon toxicity are discussed and the literature reviewed. As long as the musculoskeletal toxicity cannot be predicted by in vitro or in vivo models and this class of antibiotics is one of the most commonly linked to selection of resistant bacteria, a more prudent use of fluoroquinolones is warranted. PMID- 15794722 TI - The safety of non-stimulant agents for the treatment of attention-deficit hyperactivity disorder. AB - Due to their well-established efficacy and safety, stimulants are the drugs of first choice if medication for attention-deficit hyperactivity disorder (ADHD) is required. Nevertheless, for some individuals other, non-stimulant treatments are needed for several reasons. If so, atomoxetine is recommended as a second-line treatment. In addition, several tricyclic antidepressants, such as desipramine or imipramine, as well as alpha-2 agonists, especially clonidine or bupropion, might be efficient in treating ADHD, in particular in specific co-morbid conditions. Despite the fact that non-stimulant treatments in ADHD are usually well-tolerated with side effects being mostly moderate and transient, special safety aspects and precautions, specific for each drug, have to be considered whenever a non stimulant treatment is chosen. This review focuses on the tolerability, occurrence of adverse events, precautions required to prevent severe adverse events, and essential pharmacological interaction in the treatment of ADHD symptoms by non-stimulants. PMID- 15794723 TI - The safety of antiretroviral drugs in pregnancy. AB - Advances in HIV therapy and mother-to-child transmission (MTCT) prophylaxis have led to increasing use of antiretroviral drugs in pregnancy. Highly active antiretroviral therapy in pregnancy has been associated with prematurity, pre eclampsia and gestational diabetes. Women may be at increased risk of nevirapine associated hepatotoxicity but whether or not pregnancy is an additional risk is uncertain. Although animal studies suggest a possibility of congenital abnormalities with specific antiretrovirals, such as efavirenz, results from registries and cohort studies do not support an excess of congenital malformations associated with in utero antiretroviral exposure. Concerns regarding the health of uninfected, antiretroviral-exposed children include the potential for cancers, mitochondrial disease and haematological abnormalities. However, the absence of any excess mortality in large observational cohort studies of uninfected, antiretroviral therapy-exposed children born to HIV infected women is reassuring. Based on current knowledge, the immense benefits of antiretroviral prophylaxis in reducing the risk of MTCT, far outweigh the potential for adverse effects. PMID- 15794724 TI - Selective serotonin re-uptake inhibitors and the risk of bleeding. AB - Despite their safety, selective serotonin re-uptake inhibitors (SSRIs) are associated with bleeding. The authors critically reviewed the medical literature on SSRIs to identify subgroups of patients at risk of bleeding complications. The authors performed a literature search using MEDLINE from 1966 to 1st September 2004 using; 'haemorrhage, serotonin uptake inhibitors and antidepressive agents' as search terms and followed up on citations in each paper that was relevant to SSRI associated bleeding. The authors reviewed 7 retrospective analytical studies and 24 case reports of bleeding in 43 different people. Analytical studies support an association between SSRI consumption and upper gastrointestinal (GI) bleeding and perioperative bleeding. Little evidence links SSRI use with intracerebral haemorrhage. The risk of GI bleeding appeared to be highest among patients consuming SSRIs with NSAIDs. Combining aspirin or NSAIDs with SSRIs may further increase the risk of bleeding. Clinicians should caution patients about combining SSRIs with aspirin or NSAIDs. Pharmacotherapy to reduce the risk of GI bleeding should be considered in high risk patients. PMID- 15794725 TI - Teratogenicity of sodium valproate. AB - The teratogenicity of the widely popular antiepileptic drug (AED) and mood stabiliser sodium valproate (also known as valproate, VPA) has been evidenced by previous research; however, these findings have often been limited by a small population sample of exposed women and a retrospective study design. Many factors contribute to the teratogenicity of VPA. These include the number of drugs that are co-administered, drug dosage, differences in maternal and/or infant metabolism, the gestational age of the fetus at exposure, and hereditary susceptibility. VPA has been associated with a variety of major and minor malformations, including a 20-fold increase in neural tube defects, cleft lip and palate, cardiovascular abnormalities, genitourinary defects, developmental delay, endocrinological disorders, limb defects, and autism. It has been suggested that polytherapy treatment in epileptic pregnant women increases the risk of teratogenicity in offspring. Furthermore, there is an established relationship between VPA dose and adverse outcome. Large single doses of VPA potentially cause high peak levels in the fetal serum resulting in deleterious effects. Currently there is an increase in the number of national and international pregnancy registries being formed in an effort to better identify the teratogenic effects of AEDs. These efforts hope to enhance our understanding of AEDs and their associated risks by addressing past study limitations. PMID- 15794726 TI - Interactions between herbal and conventional medicines. AB - Herb-drug interactions are subject to much interest at present, but for various reasons reports may be unreliable or unsubstantiated. Herbal medicines are variable in composition and quality, which may affect their interaction profile as well as the reliability of reports concerning them. In this review, clinical and experimental reports have been collated, evaluated and summarised, and the theoretical and clinical evidence presented. There is an explanation of the particular issues involved with herbal medicines as compared with conventional drugs, and reasons why comparisons may or may not be valid, which is intended for those without specialist experience in herbal products. It has become apparent that only a few herbal drugs have so far been cited in interaction reports, for example St John's Wort, Ginkgo biloba, Dan Shen, liquorice, Ma huang and garlic, and that the main drugs involved are those which are already susceptible to interactions with many other conventional drugs, such as warfarin, protease inhibitors and anti-cancer drugs. An attempt has been made to put the matter into perspective and recommendations have been given for health professionals to advise or develop strategies to safeguard patients, without resorting to speculation or scaremongering. PMID- 15794727 TI - Efficacy and costs of secondary prevention with antiplatelets after ischaemic stroke. AB - Ischaemic stroke and other atherothrombotic events substantially increase the medico-economic burden because of their high treatment costs and long-lasting disabilities with need for chronic care. Studies have shown that the cost of stroke represents approximately 3 - 5% of the annual health budget. Antiplatelet agents play a major role in secondary stroke prevention. Acetylsalicylic acid (ASA), ASA combined with extended-release dipyridamole (ER-Dip), and clopidogrel are all acceptable choices for first-line treatment in the secondary prevention of stroke. The newer antiplatelets, however, are more expensive than ASA, and their cost-effectiveness is not easily estimated. ASA has to be given to 33 stroke patients to prevent one future stroke, myocardial infarction (MI) or vascular death compared with placebo. Adding ER-Dip to ASA increases the benefit for the patients. A total of 33 stroke patients had to be treated with this combination, instead of ASA, to prevent one stroke. However, the combination of ASA plus ER-Dip does not prevent MI, vascular death or the combined end point of either stroke or death. Clopidogrel is more effective than ASA in preventing a combined end point of ischaemic stroke, MI, or vascular death, but it has not been shown to be superior to ASA in preventing recurrent stroke in transient ischaemic attack or stroke patients. Several subgroups, such as stroke patients with additional peripheral artery disease, patients with prior coronary artery bypass, patients with insulin-dependent diabetes, and patients with recurrent vascular events, were identified, in whom the benefit of clopidogrel is amplified. Taking economical aspects into account, the fixed combination of ASA and ER-Dip can be recommended for secondary stroke prevention as a first-line alternative to ASA in patients without major comorbidity. In patients with higher comorbidity, clopidogrel may be more effective for the individual patient compared with ASA, and might also be cost-effective. Furthermore, in patients with ASA intolerance clopidogrel is a useful, but expensive, alternative. PMID- 15794728 TI - The economic burden of depression in the US: societal and patient perspectives. AB - Depression imposes significant costs on sufferers, their families and care givers, employers and insurance payers. This article summarises medical and health economics literature regarding the societal economic burden of depression and the incremental economic burden of depression sufferers in comparison with non-depressed counterparts. This substantial knowledge base probably underestimates the true economic burden of the disease because the available data and analysis techniques do not capture all of the subtle costs of this condition. Future investigation is likely to focus on the relationship between depression and comorbid conditions, the role of caregiver burdens, and the economic differences in life outcomes. PMID- 15794729 TI - Therapy of porphyria cutanea tarda. AB - Porphyria cutanea tarda (PCT) is the most common type of porphyria. There is an association of PCT with haemochromatosis, diabetes mellitus and hepatitis C infection. The basis of treatment of PCT consists of three elements: avoidance of triggering factors, iron depletion and porphyrin elimination. Alcohol and certain systemic medical drugs, such as oestrogens (or tuberculostatics), should be considered as triggering factors, and as far as possible, avoided. Other triggering factors, such as chronic haemodialysis in renal insufficiency, need a different approach. The hallmark in iron depletion is phlebotomy. Porphyrin elimination is achieved using low-dose chloroquin therapy. The treatment is safe and effective but has its limits in cases with haemochromatosis (HFE) gene mutations. Here iron depletion needs additional phlebotomy. In patients with chronic haemodialysis-associated PCT, chloroquine is ineffective. Erythropoietin, desferroxamine and small-volume phlebotomy have been employed to control the disease. Childhood PCT is very rare. No controlled studies are available, but published experience suggests that body weight-adapted chloroquine therapy or small volume phlebotomy might be useful. PMID- 15794730 TI - Pharmacological management of paracoccidioidomycosis. AB - A systemic and endemic emerging mycosis in Latin America, paracoccidioidomycosis, is characterised by its chronicity and by the severity of the disseminated form in healthy individuals, as well as in immunocompromised individuals co-infected with HIV, resulting, in the latter, in a mortality rate in the range of 30 - 45%. The long (several years) duration of treatment results from the immunosuppression induced by the disease or from the survival capacity of the fungus in tissue. A few controlled studies and case reports have shown that fast-acting azolic and sulfa derivatives are useful treatment alternatives for patients presenting milder forms of the disease. However, when using such drugs, treatment regimens of longer duration are required for the maintenance of patients with more severe forms. The search for new alternatives for treating the most severe forms is an ongoing challenge. Novel treatments may be found among new classes of drugs, drug combinations, or agents capable of modulating the immune response, such as a peptide derived from the 43-kDa Paracoccidioides brasiliensis glycoprotein. PMID- 15794731 TI - Treatment of hepatitis C: critical appraisal of the evidence. AB - Chronic hepatitis C virus infection is currently the most common cause of end stage liver disease worldwide. Although the conclusions of the last National Institutes of Health Consensus Development Conferences on Hepatitis C have recently been published, several important issues remain unanswered. This paper reviews the available data using an evidence-based approach. Current evidence is sufficient to recommend IFN treatment for all patients with acute hepatitis. A later initiation of therapy yields the same likelihood of response as early treatment. A daily induction dose during month 1 is the best treatment option. The current gold standard of efficacy for treatment-naive patients with chronic hepatitis C is the combination of pegylated IFN and ribavirin. The overall sustained viral response rate to these regimens is 54 - 56% following a 48-week course of therapy. Patients with genotype 1 infection will have a 42 - 51% likelihood of response to 48weeks of therapy. Those with genotypes 2 or 3 infection will respond to 24weeks in 78 - 82% of cases. Debate continues regarding the optimal dose and duration of peginterferon (PEG-IFN), not only in patients infected with genotype 2 or 3 but also in those infected with genotype 1. The optimal dose of ribavirin has yet to be determined. Available data show the need to give the highest tolerable doses (1000-1200mg/day) to the difficult to-treat patients (genotype 1, cirrhotics, obese), although there is a greater likelihood of intolerance. Genotypes 2 and 3 may receive 800mg/day, which is also the most appropriate lower dose for those patients who require dosage modification for anaemia or other side effects. Tolerability and compliance to therapy are still a problem, as approximately 15- 20% of patients within trials and > 25% in clinical practice withdraw from therapy. New PEG-IFNs are more effective than conventional IFN in improving liver histology. Monotherapy with PEG-IFN induces a marked reduction in staging in virological sustained responders, and to a lesser degree in relapsers, but provides no benefit to nonresponders after 24-48weeks of treatment. The use of maintenance therapy in virological nonresponders aiming to improve histology should be considered experimental and of unproven benefit. Pooling data from the literature suggests a slight preventive effect of IFN on hepatocellular carcinoma development in patients with HCV-related cirrhosis. The magnitude of this effect is low and the observed benefit may be due to spurious associations. The preventive effect is more evident among sustained responders to IFN. PMID- 15794732 TI - Acne vulgaris: a review of antibiotic therapy. AB - Antibiotic therapy has been integral to the management of inflammatory acne vulgaris for many years. Systemic antibiotics work via antibacterial, anti inflammatory and immunomodulatory modes of action, and have been found to be useful in managing moderate-to-severe acne. Commonly prescribed antibiotics include tetracyclines, erythromycin and trimethoprim, with or without sulfamethoxazole. In selecting the appropriate antibiotic for patients needing to receive topical or systemic antibiotic therapy, the clinician should take into account the severity of the acne, cost-effectiveness, the safety profile of the drug and the potential for development of resistance. The widespread and long term use of antibiotics over the years has unfortunately led to the emergence of resistant bacteria. The global increase in the antibiotic resistance of Propionibacterium acnes may be a significant contributing factor in treatment failures. It is therefore essential that clinicians prescribing antibiotics for the treatment of acne adopt strategies to minimise further development of bacterial resistance. This includes addressing compliance issues, using combination therapies, avoiding prolonged antibiotic treatment, and avoiding concomitant topical and oral antibiotics with chemically dissimilar antibiotics. PMID- 15794733 TI - Pharmacological treatment of disabling tremor. AB - Tremor is often a disabling primary condition or secondary to another disorder. No universally effective pharmacological agent exists for the treatment of essential tremor, and patients differ greatly in their response to therapies, thus requiring individualised regimens. Deep brain stimulation is the best option for patients with disabling, drug-resistant essential tremor. Resting tremor in Parkinson's disease is usually not the primary disabling feature, and in most cases, levodopa/carbidopa is satisfactory for many years. Young Parkinson's patients with dominant, disabling tremor benefit from anticholinergics in addition to dopaminergic therapies. However, older Parkinson's patients with more disabling tremor may suffer from dose-dependent side effects, and deep brain stimulation should be considered. This article outlines the available pharmacological agents and treatment considerations for various disabling tremors, including essential tremor and Parkinson's disease. PMID- 15794734 TI - The use of darbepoetin alfa for the treatment of chemotherapy-induced anaemia. AB - Chemotherapy-induced anaemia, with its important consequences on quality of life and social function of cancer patients, can be improved with erythropoietic therapy. Darbepoetin alfa is the first of a novel generation of erythropoietic proteins with a unique molecular structure and a circulating half-life that is threefold longer than that of the previous recombinant human erythropoietin. The efficacy and safety of weekly administration have been confirmed in different Phase II and III randomised trials. In order to optimise the efficacy profile of darbepoetin alfa, extended dosing intervals and front-loading regimens are evaluated, as well the optimal haemoglobin level to initiate therapy. Across all trials, darbepoetin alfa was shown to be a well-tolerated and safe therapy. The possible favourable effect on the outcome of cancer patients needs to be further elucidated. PMID- 15794735 TI - Cinacalcet HCl: a novel therapeutic for hyperparathyroidism. AB - Hyperparathyroidism (HPT) is a significant clinical concern for patients with a variety of diseases, notably the secondary HPT associated with chronic kidney disease requiring dialysis. Secondary HPT is associated with elevated para thyroid hormone (PTH) levels, decreased levels of 1,25 dihydroxyvitamin D, and disordered mineral levels (usually high calcium and phosphorus). If not controlled, secondary HPT can result in bone disease, vascular calcification, and ultimately, patient mortality. Established, conventional therapies, such as 1,25dihydroxyvitamin D analogues (vitamin D analogues) and phosphate binders, have proven to be inadequate in enabling patients to meet the National Kidney Foundation's-Kidney Disease Outcomes Quality Initiative (NKF-K/DOQI) treatment goals for PTH, calcium and phosphorus levels. A novel therapeutic, cinacalcet HCl (formerly AMG 073; Sensipar in the US and Mimpara in Europe; Amgen, Inc.), binds directly to the calcium-sensing receptor (CaR) on the cells of the parathyroid gland, increasing the receptor's sensitivity to calcium and reducing PTH, serum calcium and phosphorus levels. Treatment with cinacalcet in clinical trials has safely and effectively improved achievement of the NKF-K/DOQI goals. Cinacalcet has also reduced serum calcium levels in patients with primary HPT, including parathyroid carcinoma, in the clinical trial setting. Evidence suggesting the utility of cinacalcet in these diseases and the potential for additional therapeutic applications will be discussed. PMID- 15794736 TI - Enfuvirtide: the first HIV fusion inhibitor. AB - Highly active antiretroviral therapy, a combination of antiretrovirals to treat HIV-infected individuals, may fail for a number of reasons, including the selection of genetic mutations which confer resistance to the antiretroviral drugs, and poor adherence or treatment discontinuation resulting from drug toxicity. Treatment-experienced patients, who have failed therapy owing to the emergence of drug-resistant virus, have a significant unmet medical need. Enfuvirtide (T-20), the first of a new class of antiretroviral agents known as HIV fusion inhibitors, has a unique mechanism of action involving disruption of HIV entry at the stage of membrane fusion. The potent antiviral activity and favourable safety and tolerability profile of enfuvirtide has been demonstrated in combination with other agents. Its novel mechanism of action offers a low potential for cross-resistance with conventional classes of antiretrovirals, and its extracellular distribution means that drug interactions and intracellular metabolic disturbances are unlikely. Targeting viral fusion or entry will hopefully provide respite for patients who have limited treatment options following the emergence of multi-drug resistance. PMID- 15794737 TI - Risedronate for prevention and treatment of osteoporosis in postmenopausal women. AB - Risedronate sodium is an N-containing bisphosphonate that has been approved for the prevention and treatment of osteoporosis in postmenopausal women. An increase in the rate of bone remodelling is a regular feature of oestrogen withdrawal during the menopausal transition, but excessive remodelling leads to bone fragility. Risedronate and similar compounds reduce the rate of bone remodelling by suppressing the action of osteoclasts. The antifracture efficacy of risedronate is impressive. In large clinical trials of postmenopausal women with osteoporosis-related fracture(s) at entry, the risk of incident vertebral and non vertebral fractures was reduced by approximately 40%. In older women at risk for hip fracture, incident hip fractures were also reduced by approximately 40%. Antifracture efficacy develops within the first 6 months, and treatment has been followed for as long as 5 years without deleterious effects on bone. We await reports of experience with risedronate in 'real-world' cases of greater complexity (i.e., in patients with co-morbidities and medications that would have excluded them from published clinical trials). PMID- 15794738 TI - Sirolimus: a new option in transplantation. AB - This review of immunosuppression in renal transplantation has allowed us to highlight the deleterious effect of calcineurin inhibitor nephrotoxicity and to emphasise the importance of sirolimus. Now, a whole new set of possibilities has opened up in immunosuppression: sirolimus-based immunosuppression without calcineurin inhibitors; sirolimus in combination with calcineurin inhibitors in reduced doses; early calcineurin inhibitor withdrawal from a regimen that combines sirolimus, calcineurin inhibitors and steroids; and calcineurin inhibitor conversion to sirolimus when the first signs of graft nephrotoxicity appear. These new strategies in immunosuppression in renal transplantation are associated with good results in graft and patient survival in year 1, and with better renal function. Therefore, we can hope for better long-term results in transplantation, with a significant increase in the graft half-life and in the patient survival. PMID- 15794739 TI - The first 5 years of clinical experience with laronidase enzyme replacement therapy for mucopolysaccharidosis I. AB - Mucopolysaccharidosis I (MPS I, McKusick 25280) is caused by the deficiency or absence of the lysosomal enzyme, alpha-L-iduronidase (EC 3.2.1.76). This inherited disease causes progressive cellular, tissue and organ damage across the entire phenotypic spectrum. Disabling, multi-organ disease is the rule, and generally results in death between the first and fourth decades of life. Recently, laronidase (Aldurazyme) [Genzyme], a specific recombinant human alpha-L iduronidase) became commercially available as long-term enzyme replacement therapy. Results from the Phase I/II and III extended clinical studies have shown that laronidase safely and effectively alleviates many systemic signs and symptoms of this progressive multisystemic disease. Clinically meaningful and sustained improvements in pulmonary function and functional capacity have been observed in Phase III study patients. Significant and sustained reductions in urinary glysosaminoglycan (GAG) excretion and hepatomegaly have also been observed. Improvements in sleep apnoea and joint range of motion occurred in patients with the most severe symptoms at baseline. Improvements in Disability Index scores as measured using the CHAQ and HAQ questionnaires were modest, which may have been related to the fact that these disability measuring tools are not disease-specific. Anecdotal reports of improvements in the performance of daily activities further add to the therapeutic benefits, as do case histories pointing at stabilisation or improvement of symptomatology in various organs, such as the eyes, heart, and muscles. With the availability of specific treatment, the importance of early recognition of the disease and appropriate therapeutic intervention has increased. The variability in clinical symptomatology is reviewed in detail and may allow for a better understanding of the diagnostic and therapeutic challenges. Results of the clinical trials and their initial extension periods, as well as the anecdotal experiences of physicians with laronidase in non-study settings, are discussed. PMID- 15794740 TI - VALIANT (VALsartan In Acute myocardial iNfarcTion) trial. AB - Angiotensin-converting enzyme inhibitors (ACE-Is) are an evidence-based treatment for patients who after myocardial infarction (MI) present with either heart failure (HF) or left ventricular systolic dysfunction, or both. An alternative could be a more complete inhibition of the renin-angiotensin system through the blockade of the angiotensin type 1 receptors. The effect of valsartan or captopril, or the combination of the two in post-MI HF or systolic dysfunction or both, has been evaluated in the VALIANT (VALsartan In Acute myocardial iNfarcTion) trial. Total mortality and the combined secondary end point of cardiovascular death, MI or HF were not significantly different in the three groups after 24.7 months of follow-up. Valsartan was not inferior to captopril in terms of total mortality and cardiovascular death, MI and HF. Valsartan can be considered an alternative treatment to ACE-I in these patients. PMID- 15794741 TI - CHARMed - the effects of candesartan in heart failure. PMID- 15794742 TI - The therapeutic potential of vasopressin in cardiopulmonary resuscitation. PMID- 15794743 TI - Observation of a tetrahedral reaction intermediate in the HIV-1 protease substrate complex. AB - HIV-1 protease is an effective target for the design of drugs against AIDS. To help this process of drug design, three-dimensional structures have been determined of complexes between HIV-1 protease and a variety of transition-state analogue inhibitors. The true transition state, however, has not been structurally characterized. The crystal structure of the C95M/C1095A HIV-1 protease tethered dimer shows a distinctive feature in which the two flaps of the enzyme are in a 'closed conformation' even in the unliganded state. This unique feature has been utilized here to study the structure of HIV-1 protease complexed to an oligopeptide substrate of amino acid sequence His-Lys-Ala-Arg-Val-Leu*NPhe Glu-Ala-Nle-Ser (where * denotes the cleavage site, and NPhe and Nle denote p nitrophenylalanine and norleucine residues respectively). The X-ray structure of the complex refined against 2.03 A (0.203 nm) resolution synchrotron data shows that the substrate is trapped as a tetrahedral reaction intermediate in the crystal. The hydrogen-bonding interactions between the reaction intermediate and the catalytic aspartates are different from those observed previously using transition-state analogues. The reaction intermediate did not dissociate to release the products, possibly due to the inflexibility introduced in the flaps when the enzyme is packed inside crystals. PMID- 15794744 TI - Identification and characterization of novel PKA holoenzymes in human T lymphocytes. AB - Cyclic AMP-dependent protein kinase (PKA) is a holoenzyme that consists of a regulatory (R) subunit dimer and two catalytic (C) subunits that are released upon stimulation by cAMP. Immunoblotting and immunoprecipitation of T-cell protein extracts, immunofluorescence of permeabilized T cells and RT/PCR of T cell RNA using C subunit-specific primers revealed expression of two catalytically active PKA C subunits C alpha1 (40 kDa) and C beta2 (47 kDa) in these cells. Anti-RI alpha and Anti-RII alpha immunoprecipitations demonstrated that both C alpha1 and C beta2 associate with RI alpha and RII alpha to form PKAI and PKAII holoenzymes. Moreover, Anti-C beta2 immunoprecipitation revealed that C alpha1 coimmunoprecipitates with C beta2. Addition of 8-CPT-cAMP which disrupts the PKA holoenzyme, released C alpha1 but not C beta2 from the Anti-C beta2 precipitate, indicating that C beta2 and C alpha1 form part of the same holoenzyme. Our results demonstrate for the first time that various C subunits may colocate on the same PKA holoenzyme to form novel cAMP-responsive enzymes that may mediate specific effects of cAMP. PMID- 15794745 TI - Double-stranded RNA-dependent protein kinase (PKR) is downregulated by phorbol ester. AB - The double-stranded RNA-dependent protein kinase (PKR) is one of the key mediators of interferon (IFN) action against certain viruses. PKR also plays an important role in signal transduction and immunomodulation. Understanding the regulation of PKR activity is important for the use of PKR as a tool to discover and develop novel therapeutics for viral infections, cancer and immune dysfunction. We found that phorbol 12-myristate 13-acetate (PMA), a potent activator of protein kinase C (PKC), decreased the level of autophosphorylated PKR in a dose- and time-dependent manner in IFN-treated mouse fibroblast cells. Polyinosinic-polycytidylic acid (poly I:C) treatment enhanced the activity of PKR induced by IFN, but did not overcome the PMA-induced reduction of PKR autophosphorylation. Western blot analysis with a monoclonal antibody to mouse PKR revealed that the decrease of PKR autophosphorylation in cells by PMA was a result of PKR protein degradation. Selective PKC inhibitors blocked the degradation of PKR stimulated by PMA, indicating that PKC activity was required for the effect. Furthermore, we also found that proteasome inhibitors prevented PMA-induced down regulation of PKR, indicating that an active proteasome is required. Our results identify a novel mechanism for the post-translational regulation of PKR. PMID- 15794746 TI - Interactions of ultraspiracle with ecdysone receptor in the transduction of ecdysone- and juvenile hormone-signaling. AB - Analyses of integration of two-hormone signaling through the vertebrate nuclear hormone receptors, for which the retinoid X receptor is one partner, have generated a number of mechanistic models, including those described as 'subordination' models wherein ligand-activation of one partner is subordinate to the liganded state of the other partner. However, mechanisms by which two-hormone signaling is integrated through invertebrate nuclear hormone-binding receptors has not been heretofore experimentally elucidated. This report investigates the integration of signaling of invertebrate juvenile hormone (JH) and 20-OH ecdysone (20OHE) at the level of identified nuclear receptors (ultraspiracle and ecdysone receptor), which transcriptionally activate a defined model core promoter (JH esterase gene), through specified hormone response elements (DR1 and IR1). Application of JH III, or 20OHE, to cultured Sf9 cells transfected with a DR1JHECoreLuciferase (or IR1JHECoreLuciferase) reporter promoter each induced expression of the reporter. Cotreatment of transfected cells with both hormones yielded a greater than additive effect on transcription, for especially the IR1JHECoreLuciferase reporter. Overexpression in Sf9 cells of recombinant Drosophila melanogaster ultraspiracle (dUSP) fostered formation of dUSP oligomer (potentially homodimer), as measured by coimmunoprecipitation assay and electrophoretic mobility assay (EMSA) on a DR1 probe, and also increased the level of transcription in response to JH III, but did not increase the transcriptional response to either 20OHE treatment alone or to the two hormones together. Inapposite, overexpression of recombinant D. melanogaster ecdysone receptor (dEcR) in the transfected cells generated dUSP/dEcR heterodimer [as measured by EMSA (supershift) on a DR1 probe] and increased the transcriptional response to 20OHE-alone treatment, but did not increase the transcriptional response to the JH III-alone treatment. Our studies provide evidence that in this model system, JH III-activation of the reporter promoter is through USP oligomer (homodimer) that does not contain EcR, while the 20OHE-activation is through the USP/EcR heterodimer. These results also show that the integration of JH III and 20OHE signaling is through the USP/EcR heterodimer, but that when the EcR partner is unliganded, the USP partner in this system is unable to transduce the JH III activation. PMID- 15794747 TI - SLC39A14, a LZT protein, is induced in adipogenesis and transports zinc. AB - During adipocyte differentiation, there is an underlying complex series of gene expressions. We have previously isolated many genes whose expression levels are quickly elevated by the addition of inducers to mouse 3T3-L1 preadipocyte cells. Here we report the isolation and characterization of SLC39A14, a member of the LZT proteins, one of the subfamilies of ZIP transporters. The expression of the SLC39A14 gene was strongly and rapidly induced at the early stages of differentiation. Moreover, it was highly restricted to the potential differentiation state of 3T3-L1 cells and the expression level was quite low in the nonadipogenic NIH-3T3 cells, indicating a dominant expression in adipocyte differentiation. The zinc uptake assay revealed that SLC39A14 functions as a zinc transporter. Taken together, these results suggest that SLC39A14 plays a role as a zinc transporter during the early stages of adipogenesis. PMID- 15794748 TI - Novel target genes of the Wnt pathway and statistical insights into Wnt target promoter regulation. AB - The Wnt pathway controls biological processes via the regulation of target gene expression. The expression of direct Wnt target genes, e.g. cyclin D1 and MYC, is activated by the transcription factor TCF, which binds to specific sequence motifs in the promoter. Indirect target genes are regulated via transcription regulators, which are targets of the Wnt pathway. As an example, MYC regulates the MYC interacting zinc finger protein-1 (MIZ-1), which is able to inhibit the expression of the indirect target p21WAF1. We intended to identify new Wnt target genes and to get a deeper insight into the regulatory mechanisms of Wnt target gene expression. For this we analyzed the differential expression pattern of Wnt 1 activated cells by microarray analysis. We identified 43 sequences including eight expressed sequence tags (ESTs), which showed increased transcript levels, and 104 sequences including 19 ESTs with decreased RNA levels. Northern blot and real-time quantitative PCR analysis of the differential expression levels of 15 genes confirmed the differential expression trends of eight candidate genes. When the Wnt pathway was regulated at the lower level of glycogen synthase kinase-3 beta (GSK-3 beta) or adenomatous polyposis coli (APC), we detected discrepant expression trends. We compared the number of binding sites of transcription factors in the genomic regions of all candidate target genes with the number of sites in control genes. We found that the genomic regions of the down-regulated genes include an increased number of putative MIZ-1 binding sites. Our study introduces several new Wnt target genes and provides indications that the specific gene expression pattern depends on the type of the activation trigger or the level of interference with the Wnt pathway. Furthermore, our data indicate that a high proportion of Wnt target genes are regulated by indirect mechanisms. PMID- 15794749 TI - Training of yeast cell dynamics. AB - In both industrial fermenters and in their natural habitats, microorganisms often experience an inhomogeneous and fluctuating environment. In this paper we mimicked one aspect of this nonideal behaviour by imposing a low and oscillating extracellular glucose concentration on nonoscillating suspensions of yeast cells. The extracellular dynamics changed the intracellular dynamics--which was monitored through NADH fluorescence--from steady to equally dynamic; the latter followed the extracellular dynamics at the frequency of glucose pulsing. Interestingly, the amplitude of the oscillation of the NADH fluorescence increased with time. This increase in amplitude was sensitive to inhibition of protein synthesis, and was due to a change in the cells rather than in the medium; the cell population was 'trained' to respond to the extracellular dynamics. To examine the mechanism behind this 'training', we subjected the cells to a low and constant extracellular glucose concentration. Seventy-five minutes of adaptation to a low and constant glucose concentration induced the same increase of the amplitude of the forced NADH oscillations as did the train of glucose pulses. Furthermore, 75 min of adaptation to a low (oscillating or continuous) glucose concentration decreased the K(M) of the glucose transporter from 26 mm to 3.5 mm. When subsequently the apparent K(M) was increased by addition of maltose, the amplitude of the forced oscillations dropped to its original value. This demonstrated that the increased affinity of glucose transport was essential for the training of the cells' dynamics. PMID- 15794750 TI - The role of N-glycosylation in the stability, trafficking and GABA-uptake of GABA transporter 1. Terminal N-glycans facilitate efficient GABA-uptake activity of the GABA transporter. AB - Neurotransmitter transporters play a major role in achieving low concentrations of their respective transmitter in the synaptic cleft. The GABA transporter GAT1 belongs to the family of Na(+)- and Cl(-)-coupled transport proteins which possess 12 putative transmembrane domains and three N-glycosylation sites in the extracellular loop between transmembrane domain 3 and 4. To study the significance of N-glycosylation, green fluorescence protein (GFP)-tagged wild type GAT1 (NNN) and N-glycosylation defective mutants (DDQ, DGN, DDN and DDG) were expressed in CHO cells. Compared with the wild type, all N-glycosylation mutants showed strongly reduced protein stability and trafficking to the plasma membrane, which however were not affected by 1-deoxymannojirimycin (dMM). This indicates that N-glycosylation, but not terminal trimming of the N-glycans is involved in the attainment of a correctly folded and stable conformation of GAT1. All N-glycosylation mutants were expressed on the plasma membrane, but they displayed markedly reduced GABA-uptake activity. Also, inhibition of oligosaccharide processing by dMM led to reduction of this activity. Further experiments showed that both N-glycosylation mutations and dMM reduced the V(max) value, while not increasing the K(m) value for GABA uptake. Electrical measurements revealed that the reduced transport activity can be partially attributed to a reduced apparent affinity for extracellular Na+ and slowed kinetics of the transport cycle. This indicates that N-glycans, in particular their terminal trimming, are important for the GABA-uptake activity of GAT1. They play a regulatory role in the GABA translocation by affecting the affinity and the reaction steps associated with the sodium ion binding. PMID- 15794751 TI - Exploring the GluR2 ligand-binding core in complex with the bicyclical AMPA analogue (S)-4-AHCP. AB - The X-ray structure of the ionotropic GluR2 ligand-binding core (GluR2-S1S2J) in complex with the bicyclical AMPA analogue (S)-2-amino-3-(3-hydroxy-7,8-dihydro-6H cyclohepta[d]-4-isoxazolyl)propionic acid [(S)-4-AHCP] has been determined, as well as the binding pharmacology of this construct and of the full-length GluR2 receptor. (S)-4-AHCP binds with a glutamate-like binding mode and the ligand adopts two different conformations. The K(i) of (S)-4-AHCP at GluR2-S1S2J was determined to be 185 +/- 29 nM and at full-length GluR2(R)o it was 175 +/- 8 nM. (S)-4-AHCP appears to elicit partial agonism at GluR2 by inducing an intermediate degree of domain closure (17 degrees). Also, functionally (S)-4-AHCP has an efficacy of 0.38 at GluR2(Q)i, relative to (S)-glutamate. The proximity of bound (S)-4-AHCP to domain D2 prevents full D1-D2 domain closure, which is limited by steric repulsion, especially between Leu704 and the ligand. PMID- 15794752 TI - Characterization of depolarization and repolarization phases of mitochondrial membrane potential fluctuations induced by tetramethylrhodamine methyl ester photoactivation. AB - Depolarization and repolarization phases (D and R phases, respectively) of mitochondrial potential fluctuations induced by photoactivation of the fluorescent probe tetramethylrhodamine methyl ester (TMRM) were analyzed separately and investigated using specific inhibitors and substrates. The frequency of R phases was significantly inhibited by oligomycin and aurovertin (mitochondrial ATP synthase inhibitors), rotenone (mitochondrial complex I inhibitor) and iodoacetic acid (inhibitor of the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase). Succinic acid (mitochondrial complex II substrate, given in the permeable form of dimethyl ester) abolished the rotenone-induced inhibition of R phases. Taken together, these findings indicate that the activity of both respiratory chain and ATP synthase were required for the recovery of the mitochondrial potential. The frequency of D phases prevailed over that of R phases in all experimental conditions, resulting in a progressive depolarization of mitochondria accompanied by NAD(P)H oxidation and Ca2+ influx. D phases were not blocked by cyclosporin A (inhibitor of the permeability transition pore) or o phenyl-EGTA (a Ca2+ chelator), suggesting that the permeability transition pore was not involved in mitochondrial potential fluctuations. PMID- 15794753 TI - Synthesis of beta-mannosides using the transglycosylation activity of endo-beta mannosidase from Lilium longiflorum. AB - Endo-beta-mannosidase is an endoglycosidase that hydrolyzes only the Man beta 1 4GlcNAc linkage of the core region of N-linked sugar chains. Recently, endo-beta mannosidase was purified to homogeneity from Lilium longiflorum (Lily) flowers, its corresponding gene was cloned and important catalytic amino acid residues were identified [Ishimizu T., Sasaki A., Okutani S., Maeda M., Yamagishi M. & Hase S. (2004) J. Biol. Chem.279, 38555-38562]. In the presence of Man beta 1 4GlcNAc beta 1-4GlcNAc-peptides as a donor substrate and p-nitrophenyl beta-N acetylglucosaminide as an acceptor substrate, the enzyme transferred mannose to the acceptor substrate by a beta1-4-linkage regio-specifically and stereo specifically to give Man beta 1-4GlcNAc beta 1-pNP as a transfer product. Further studies indicated that not only p-nitrophenyl beta-N-acetylglucosaminide but also p-nitrophenyl beta-glucoside and p-nitrophenyl beta-mannoside worked as acceptor substrates, however, p-nitrophenyl beta-N-acetylgalactosaminide did not work, indicating that the configuration of the hydroxyl group at the C4 position of an acceptor is important. Besides mannose, oligomannoses were also transferred. In the presence of (Man)(n)Man alpha 1-6Man beta 1-4GlcNAc beta 1-4GlcNAc-peptides (n = 0-2) and pyridylamino GlcNAc beta 1-4GlcNAc, the enzyme transferred (Man)(n)Man alpha 1-6Man en bloc to the acceptor substrate to produce pyridylamino (Man)(n)Man alpha 1-6Man beta 1-4GlcNAc beta 1-4GlcNAc (n =0-2). Thus, the lily endo-beta-mannosidase is useful for the enzymatic preparation of oligosaccharides containing the mannosyl beta 1,4-structure, chemical preparations of which have been frequently reported to be difficult. PMID- 15794754 TI - Role of phosphorylation in p53 acetylation and PAb421 epitope recognition in baculoviral and mammalian expressed proteins. AB - Post-translational modifications, such as phosphorylation and acetylation of the tumour suppressor protein p53, elicit important effects on the function and the stability of the resultant protein. However, as phosphorylation and acetylation are dynamic events subject to complex controls, elucidating the relationships between phosphorylation and acetylation is difficult. In the present study we sought to address this problem by comparing full-length wild-type p53 with full length p53 proteins mutated at specific phosphorylation targets. Recombinant murine p53 proteins were expressed in insect cells (using the baculoviral expression vector system) and in a mammalian in vitro transcription/translation reticulocyte lysate system. In p53 proteins derived from baculoviral expression vectors, S37A (but not S37D) was found to abrogate phosphorylation at S15. Lysine 382 (K382) is constitutively acetylated and was shown to form part of the epitope recognized by PAb421. Lysine 373 (K373) was only acetylated following substitutions at S315 (S315A or S315D) or at S378 (S378A). Importantly, in baculoviral expressed proteins, PAb421 reactivity was independent of K373 acetylation status, indicating that acetylation at K382 specifically determines the PAb421 epitope. PMID- 15794755 TI - The Ets transcription factor ESE-1 mediates induction of the COX-2 gene by LPS in monocytes. AB - Cyclooxygenase-2 (COX-2) is a key enzyme in the production of prostaglandins that are major inflammatory agents. COX-2 production is triggered by exposure to various cytokines and to bacterial endotoxins. We present here a novel role for the Ets transcription factor ESE-1 in regulating the COX-2 gene in response to endotoxin and other pro-inflammatory stimuli. We report that the induction of COX 2 expression by lipopolysaccharide (LPS) and pro-inflammatory cytokines correlates with ESE-1 induction in monocyte/macrophages. ESE-1, in turn, binds to several E26 transformation specific (Ets) sites on the COX-2 promoter. In vitro analysis demonstrates that ESE-1 binds to and activates the COX-2 promoter to levels comparable to LPS-mediated induction. Moreover, we provide results showing that the induction of COX-2 by LPS may require ESE-1, as the mutation of the Ets sites in the COX-2 promoter or overexpression of a dominant-negative form of ESE 1 inhibits LPS-mediated COX-2 induction. The effect of ESE-1 on the COX-2 promoter is further enhanced by cooperation with other transcription factors such as nuclear factor-kappa B and nuclear factor of activated T cells. Neutralization of COX-2 is the goal of many anti-inflammatory drugs. As an activator of COX-2 induction, ESE-1 may become a target for such therapeutics as well. Together with our previous reports of the role of ESE-1 as an inducer of nitric oxide synthase in endothelial cells and as a mediator of pro-inflammatory cytokines in vascular and connective tissue cells, these results establish ESE-1 as an important player in the regulation of inflammation. PMID- 15794756 TI - Bioinformatic and enzymatic characterization of the MAPEG superfamily. AB - The membrane associated proteins in eicosanoid and glutathione metabolism (MAPEG) superfamily includes structurally related membrane proteins with diverse functions of widespread origin. A total of 136 proteins belonging to the MAPEG superfamily were found in database and genome screenings. The members were found in prokaryotes and eukaryotes, but not in any archaeal organism. Multiple sequence alignments and calculations of evolutionary trees revealed a clear subdivision of the eukaryotic MAPEG members, corresponding to the six families of microsomal glutathione transferases (MGST) 1, 2 and 3, leukotriene C4 synthase (LTC4), 5-lipoxygenase activating protein (FLAP), and prostaglandin E synthase. Prokaryotes contain at least two distinct potential ancestral subfamilies, of which one is unique, whereas the other most closely resembles enzymes that belong to the MGST2/FLAP/LTC4 synthase families. The insect members are most similar to MGST1/prostaglandin E synthase. With the new data available, we observe that fish enzymes are present in all six families, showing an early origin for MAPEG family differentiation. Thus, the evolutionary origins and relationships of the MAPEG superfamily can be defined, including distinct sequence patterns characteristic for each of the subfamilies. We have further investigated and functionally characterized representative gene products from Escherichia coli, Synechocystis sp., Arabidopsis thaliana and Drosophila melanogaster, and the fish liver enzyme, purified from pike (Esox lucius). Protein overexpression and enzyme activity analysis demonstrated that all proteins catalyzed the conjugation of 1-chloro-2,4 dinitrobenzene with reduced glutathione. The E. coli protein displayed glutathione transferase activity of 0.11 micromol.min(-1).mg(-1) in the membrane fraction from bacteria overexpressing the protein. Partial purification of the Synechocystis sp. protein yielded an enzyme of the expected molecular mass and an N-terminal amino acid sequence that was at least 50% pure, with a specific activity towards 1-chloro-2,4-dinitrobenzene of 11 micromol.min(-1).mg(-1). Yeast microsomes expressing the Arabidopsis enzyme showed an activity of 0.02 micromol.min(-1).mg(-1), whereas the Drosophila enzyme expressed in E. coli was highly active at 3.6 micromol.min(-1).mg(-1). The purified pike enzyme is the most active MGST described so far with a specific activity of 285 micromol.min( 1).mg(-1). Drosophila and pike enzymes also displayed glutathione peroxidase activity towards cumene hydroperoxide (0.4 and 2.2 micromol.min(-1).mg(-1), respectively). Glutathione transferase activity can thus be regarded as a common denominator for a majority of MAPEG members throughout the kingdoms of life whereas glutathione peroxidase activity occurs in representatives from the MGST1, 2 and 3 and PGES subfamilies. PMID- 15794757 TI - Novel aggregate formation of a frame-shift mutant protein of tissue-nonspecific alkaline phosphatase is ascribed to three cysteine residues in the C-terminal extension. Retarded secretion and proteasomal degradation. AB - In the majority of hypophosphatasia patients, reductions in the serum levels of alkaline phosphatase activity are caused by various missense mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) gene. A unique frame-shift mutation due to a deletion of T at cDNA number 1559 [TNSALP (1559delT)] has been reported only in Japanese patients with high allele frequency. In this study, we examined the molecular phenotype of TNSALP (1559delT) using in vitro translation/translocation system and COS-1 cells transiently expressing this mutant protein. We showed that the mutant protein not only has a larger molecular size than the wild type enzyme by approximately 12 kDa, reflecting an 80 amino acid-long extension at its C-terminus, but that it also lacks a glycosylphosphatidylinositol anchor. In support of this, alkaline phosphatase activity of the cells expressing TNSALP (1559delT) was localized at the juxtanucleus position, but not on the cell surface. However, only a limited amount of the newly synthesized protein was released into the medium and the rest was polyubiquitinated, followed by degradation in the proteasome. SDS/PAGE and analysis by sucrose-density-gradient analysis indicated that TNSALP (1559delT) forms a disulfide-bonded high-molecular-mass aggregate. Interestingly, the aggregate form of TNSALP (1559delT) exhibited a significant enzyme activity. When all three cysteines at positions of 506, 521 and 577 of TNSALP (1559delT) were replaced with serines, the aggregation disappeared and instead this modified mutant protein formed a noncovalently associated dimer, strongly indicating that these cysteine residues in the C-terminal region are solely responsible for aggregate formation by cross-linking the catalytically active dimers. Thus, complete absence of TNSALP on cell surfaces provides a plausible explanation for a severe lethal phenotype of a homozygote hypophosphatasia patient carrying TNSALP (1559delT). PMID- 15794758 TI - Hu-K4 is a ubiquitously expressed type 2 transmembrane protein associated with the endoplasmic reticulum. AB - Hu-K4 is a human protein homologous to the K4L protein of vaccinia virus. Due to the presence of two HKD motifs, Hu-K4 was assigned to the family of Phospholipase D proteins although so far no catalytic activity has been shown. The Hu-K4 mRNA is found in many human organs with highest expression levels in the central nervous system. We extended the ORF of Hu-K4 to the 5' direction. As a consequence the protein is 53 amino acids larger than originally predicted, now harbouring a putative transmembrane domain. The exon/intron structure of the Hu K4 gene reveals extensive alternative splicing in the 5' untranslated region. Due to the absence of G/C-rich regions and upstream ATG codons, the mRNA isoform in brain may be translated with higher efficacy leading to a high Hu-K4 protein concentration in this tissue. Using a specific antiserum produced against Hu-K4 we found that Hu-K4 is a membrane-bound protein colocalizing with protein disulfide isomerase, a marker of the endoplasmic reticulum. Glycosylation of Hu K4 as shown by treatment with peptide N-glycosidase F or tunicamycin indicates that Hu-K4 has a type 2 transmembrane topology. PMID- 15794759 TI - Oxidative folding of conotoxins sharing an identical disulfide bridging framework. AB - Conotoxins are short, disulfide-rich peptide neurotoxins produced in the venom of predatory marine cone snails. It is generally accepted that an estimated 100,000 unique conotoxins fall into only a handful of structural groups, based on their disulfide bridging frameworks. This unique molecular diversity poses a protein folding problem of relationships between hypervariability of amino acid sequences and mechanism(s) of oxidative folding. In this study, we present a comparative analysis of the folding properties of four conotoxins sharing an identical pattern of cysteine residues forming three disulfide bridges, but otherwise differing significantly in their primary amino acid sequence. Oxidative folding properties of M-superfamily conotoxins GIIIA, PIIIA, SmIIIA and RIIIK varied with respect to kinetics and thermodynamics. Based on rates for establishing the steady-state distribution of the folding species, two distinct folding mechanisms could be distinguished: first, rapid-collapse folding characterized by very fast, but low-yield accumulation of the correctly folded form; and second, slow rearrangement folding resulting in higher accumulation of the properly folded form via the reshuffling of disulfide bonds within folding intermediates. Effects of changing the folding conditions indicated that the rapid-collapse and the slow rearrangement mechanisms were mainly determined by either repulsive electrostatic or productive noncovalent interactions, respectively. The differences in folding kinetics for these two mechanisms were minimized in the presence of protein disulfide isomerase. Taken together, folding properties of conotoxins from the M superfamily presented in this work and from the O-superfamily published previously suggest that conotoxin sequence diversity is also reflected in their folding properties, and that sequence information rather than a cysteine pattern determines the in vitro folding mechanisms of conotoxins. PMID- 15794760 TI - Neptunium uptake by serum transferrin. AB - Although of major impact in terms of biological and environmental hazards, interactions of actinide cations with biological molecules are only partially understood. Human serum transferrin (Tf) is one of the major iron carriers in charge of iron regulation in the cell cycle and consequently contamination by actinide cations is a critical issue of nuclear toxicology. Combined X-ray absorption spectroscopy (XAS) and near infrared absorption spectrometry were used to characterize a new complex between Tf and Np (IV) with the synergistic nitrilotriacetic acid (NTA) anion. Description of the neptunium polyhedron within the iron coordination site is given. PMID- 15794761 TI - Emergence of a subfamily of xylanase inhibitors within glycoside hydrolase family 18. AB - The xylanase inhibitor protein I (XIP-I), recently identified in wheat, inhibits xylanases belonging to glycoside hydrolase families 10 (GH10) and 11 (GH11). Sequence and structural similarities indicate that XIP-I is related to chitinases of family GH18, despite its lack of enzymatic activity. Here we report the identification and biochemical characterization of a XIP-type inhibitor from rice. Despite its initial classification as a chitinase, the rice inhibitor does not exhibit chitinolytic activity but shows specificities towards fungal GH11 xylanases similar to that of its wheat counterpart. This, together, with an analysis of approximately 150 plant members of glycosidase family GH18 provides compelling evidence that xylanase inhibitors are largely represented in this family, and that this novel function has recently emerged based on a common scaffold. The plurifunctionality of GH18 members has major implications for genomic annotations and predicted gene function. This study provides new information which will lead to a better understanding of the biological significance of a number of GH18 'inactivated' chitinases. PMID- 15794762 TI - Characterization of HbpR binding by site-directed mutagenesis of its DNA-binding site and by deletion of the effector domain. AB - In the presence of 2-hydroxybiphenyl, the enhancer binding protein, HbpR, activates the sigma54-dependent P(hbpC) promoter and controls the initial steps of 2-hydroxybiphenyl degradation in Pseudomonas azelaica. In the activation process, an oligomeric HbpR complex of unknown subunit composition binds to an operator region containing two imperfect palindromic sequences. Here, the HbpR DNA binding interactions were investigated by site-directed mutagenesis of the operator region and by DNA-binding assays using purified HbpR. Mutations that disrupted the twofold symmetry in the palindromes did not affect the binding affinity of HbpR, but various mutations along a 60 bp region, and also outside the direct palindromic sequences, decreased the binding affinity. Footprints of HbpR on mutant operator fragments showed that a partial loss of binding contacts occurs, suggesting that the binding of one HbpR 'protomer' in the oligomeric complex is impaired whilst leaving the other contacts intact. An HbpR variant, devoid of its N-terminal sensing A-domain, was unable to activate transcription from the hbpC promoter while maintaining protection of the operator DNA in footprints. Wild-type HbpR was unable to activate transcription from the hbpC promoter when delta A-HbpR was expressed in the same cell, suggesting the formation of (repressing) hetero-oligomers. This model implies that HbpR can self associate on its operator DNA without effector recognition or ATP binding. Furthermore, our findings suggest that the N-terminal sensing domain of HbpR is needed to activate the central ATPase domain rather than to repress a constitutively active C domain, as is the case for the related regulatory protein XylR. PMID- 15794763 TI - Glycolysis in Entamoeba histolytica. Biochemical characterization of recombinant glycolytic enzymes and flux control analysis. AB - The synthesis of ATP in the human parasite Entamoeba histolytica is carried out solely by the glycolytic pathway. Little kinetic and structural information is available for most of the pathway enzymes. We report here the gene cloning, overexpression and purification of hexokinase, hexose-6-phosphate isomerase, inorganic pyrophosphate-dependent phosphofructokinase, fructose-1,6 bisphosphate aldolase (ALDO), triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase, phosphoglycerate mutase (PGAM), enolase, and pyruvate phosphate dikinase (PPDK) enzymes from E. histolytica. Kinetic characterization of these 10 recombinant enzymes was made, establishing the kinetic constants at optimal and physiological pH values, analyzing the effect of activators and inhibitors, and investigating the storage stability and oligomeric state. Determination of the catalytic efficiencies at the pH optimum and at pH values that resemble those of the amoebal trophozoites was performed for each enzyme to identify possible controlling steps. This analysis suggested that PGAM, ALDO, GAPDH, and PPDK might be flux control steps, as they showed the lowest catalytic efficiencies. An in vitro reconstruction of the final stages of glycolysis was made to determine their flux control coefficients. Our results indicate that PGAM and PPDK exhibit high control coefficient values at physiological pH. PMID- 15794764 TI - Roles of 1-Cys peroxiredoxin in haem detoxification in the human malaria parasite Plasmodium falciparum. AB - In the present study, we investigated whether Plasmodium falciparum 1-Cys peroxiredoxin (Prx) (Pf1-Cys-Prx), a cytosolic protein expressed at high levels during the haem-digesting stage, can act as an antioxidant to cope with the oxidative burden of haem (ferriprotoporphyrin IX; FP). Recombinant Pf1-Cys-Prx protein (rPf1-Cys-Prx) competed with glutathione (GSH) for FP and inhibited FP degradation by GSH. When rPf1-Cys-Prx was added to GSH-mediated FP degradation, the amount of iron released was reduced to 23% of the reaction without the protein (P < 0.01). The rPf1-Cys-Prx bound to FP-agarose at pH 7.4, which is the pH of the parasite cytosol. The rPf1-Cys-Prx could completely protect glutamine synthetase from inactivation by the dithiothreitol-Fe(3+)-dependent mixed function oxidation system, and it also protected enolase from inactivation by coincubation with FP/GSH. Incubation of white ghosts of human red blood cells and FP with rPf1-Cys-Prx reduced formation of membrane associations with FP to 75% of the incubation without the protein (P < 0.01). The findings of the present study suggest that Pf1-Cys-Prx protects the parasite against oxidative stresses by binding to FP, slowing the rate of GSH-mediated FP degradation and consequent iron generation, protecting proteins from iron-derived reactive oxygen species, and interfering with formation of membrane-associated FP. PMID- 15794765 TI - Induction of raft-like domains by a myristoylated NAP-22 peptide and its Tyr mutant. AB - The N-terminally myristoylated, 19-amino acid peptide, corresponding to the amino terminus of the neuronal protein NAP-22 (NAP-22 peptide) is a naturally occurring peptide that had been shown by fluorescence to cause the sequestering of a Bodipy labeled PtdIns(4,5)P2 in a cholesterol-dependent manner. The present work, using differential scanning calorimetry (DSC), extends the observation that formation of a PtdIns(4,5)P2-rich domain is cholesterol dependent and shows that it also leads to the formation of a cholesterol-depleted domain. The PtdIns(4,5)P2 used in the present work is extracted from natural sources and does not contain any label and has the native acyl chain composition. Peptide-induced formation of a cholesterol-depleted domain is abolished when the sole aromatic amino acid, Tyr11 is replaced with a Leu. Despite this, the modified peptide can still sequester PtdIns(4,5)P2 into domains, probably because of the presence of a cluster of cationic residues in the peptide. Cholesterol and PtdIns(4,5)P2 also modulate the insertion of the peptide into the bilayer as revealed by 1H NOESY MAS/NMR. The intensity of cross peaks between the aromatic protons of the Tyr residue and the protons of the lipid indicate that in the presence of cholesterol there is a change in the nature of the interaction of the peptide with the membrane. These results have important implications for the mechanism by which NAP-22 affects actin reorganization in neurons. PMID- 15794766 TI - Silencing the expression of mitochondrial acyl-CoA thioesterase I and acyl-CoA synthetase 4 inhibits hormone-induced steroidogenesis. AB - Arachidonic acid and its lypoxygenated metabolites play a fundamental role in the hormonal regulation of steroidogenesis. Reduction in the expression of the mitochondrial acyl-CoA thioesterase (MTE-I) by antisense or small interfering RNA (siRNA) and of the arachidonic acid-preferring acyl-CoA synthetase (ACS4) by siRNA produced a marked reduction in steroid output of cAMP-stimulated Leydig cells. This effect was blunted by a permeable analog of cholesterol that bypasses the rate-limiting step in steroidogenesis, the transport of cholesterol from the outer to the inner mitochondrial membrane. The inhibition of steroidogenesis was overcome by addition of exogenous arachidonic acid, indicating that the enzymes are part of the mechanism responsible for arachidonic acid release involved in steroidogenesis. Knocking down the expression of MTE-I leads to a significant reduction in the expression of steroidogenic acute regulatory protein. This protein is induced by arachidonic acid and controls the rate-limiting step. Overexpression of MTE-I resulted in an increase in cAMP-induced steroidogenesis. In summary, our results demonstrate a critical role for ACS4 and MTE-I in the hormonal regulation of steroidogenesis as a new pathway of arachidonic acid release different from the classical phospholipase A2 cascade. PMID- 15794768 TI - Robotic urology in the UK: establishing a programme and emerging role. PMID- 15794769 TI - Laparoscopic reconstructive urology. PMID- 15794770 TI - Drug-eluting biomaterials in urology: the time is ripe. PMID- 15794771 TI - The European working-time directive: one step forward, two steps back. PMID- 15794772 TI - Peyronie's disease: the epidemiology, aetiology and clinical evaluation of deformity. AB - There is a wide variety of topics covered in this section. The epidemiology, aetiology and clinical evaluation of the deformity in Peyronie's disease is described, followed by a discussion of recent advances in the biology of diabetes associated bladder complications. Bladder cancer and its molecular prognostic factors are presented, and the section ends with an in-depth presentation of an evidence-based approach to the understanding of the pharmacological class effect in the management of prostatic diseases. PMID- 15794773 TI - Recent advances in understanding the biology of diabetes-associated bladder complications and novel therapy. PMID- 15794774 TI - Molecular prognostic factors in bladder cancer. PMID- 15794775 TI - An evidence-based approach to understanding the pharmacological class effect in the management of prostatic diseases. PMID- 15794776 TI - Radical prostatectomy for clinically advanced (cT3) prostate cancer since the advent of prostate-specific antigen testing: 15-year outcome. AB - In the first paper in this section, authors from the Mayo Clinic describe their experience and 15-year outcomes in the controversial subject of radical prostatectomy in patients with clinical T3 prostate cancer. The findings were interesting in many respects, but the authors concluded that radical prostatectomy as part of multimodal treatment for patients with clinical T3 disease offers cancer control and good survival rates. There follows a series of papers on both prostate cancer and bladder cancer, but the final paper in this section from the UK attempts to define the accuracy of urologists and oncologists in assessing patient life-expectancy. Using various methods they found that, rather disappointingly, doctors were poor at predicting 10-year survival, leading to the possible outcome that some patients may be denied treatment after a pessimistic assessment of life-expectancy. OBJECTIVE: To report a long-term experience with extirpative surgery in patients presenting with locally advanced (cT3) prostate cancer, as the best management of such patients remains a problem. PATIENTS AND METHODS: In a single-institution retrospective study identifying 5652 men who had radical prostatectomy (RP) for histologically confirmed prostate cancer since the advent of prostate-specific antigen (PSA) testing (1987-97), 15% (842) had RP for cT3 disease. The median follow-up of these men was 10.3 years. Cancer-specific, overall and disease-free survival was plotted and compared with those of patients having RP for cT2 disease during the same period. Perioperative morbidity, continence and erectile function rates were examined, with a multivariate analysis for risk factors of disease recurrence. RESULTS: Freedom from local or systemic disease at 5, 10, and 15 years after RP for cT3 disease was 85%, 73% and 67%; the respective cancer-specific survival rates were 95%, 90% and 79%. Significantly many men who did not receive neoadjuvant therapy (27%) were clinically over-staged (pT2) and most men with pT3 disease (78%) received adjuvant therapy. The mean time to adjuvant therapy after RP was not significantly different between men with cT3 and cT2 disease (4.0 and 4.3 years). Pathological grade (> or =7), positive surgical margins, and nondiploid chromatin were all independently associated with a significant risk for clinical disease recurrence, while preoperative PSA level had little effect on outcome. Complications and continence rates after RP in patients with cT3 mirrored those in patients with cT2 disease. CONCLUSIONS: Significantly many patients with cT3 prostate cancer are overstaged (pT2) in the PSA era. RP as part of a multimodal treatment strategy for patients with cT3 disease offers cancer control and survival rates approaching those achieved for cT2 disease. Pathological grade, ploidy and margin status are all significant predictors of outcome after RP. Complications and incontinence rates in patients with cT3 disease mirror those after RP for cT2 disease. PMID- 15794777 TI - The influence of bladder neck mucosal eversion and early urinary extravasation on patient outcome after radical retropubic prostatectomy: a prospective controlled trial. AB - OBJECTIVE: To evaluate the role of bladder neck (BN) mucosal eversion during retropubic radical prostatectomy (RRP) on the rate of BN sclerosis and urinary incontinence, with the hypothesis that BN mucosal eversion is not essential to improve the clinical outcome after RRP. PATIENTS AND METHODS: One hundred patients with stage T1c-T2c prostate cancer had RRP by the same surgeon and were randomly divided in two equal groups; one had a vesico-urethral anastomosis with and one with no BN mucosal eversion. The patients were assessed by retrograde cysto-urethrography 4 days after surgery to evaluate the presence of urinary leakage. The occurrence of BN sclerosis and the rate of urinary incontinence (more than one pad/day) was assessed by double-blind interviews at 2 days, 2 months and 6 months after catheter removal, and the incidence of BN sclerosis was also assessed after 12 months. RESULTS: In the groups with or with no BN mucosal eversion, 48 and 47 patients, respectively, fulfilled the selection criteria. Urinary leakage after vesico-urethral anastomosis was more common after mucosal eversion (33% vs 21%), but not significantly (P = 0.251). BN sclerosis occurred in only one patient, with no mucosal eversion. The rate of urinary continence was similar in both groups at 2 days (69% vs 68%, respectively), 2 months (90% vs 87%) and 6 months (92% vs 92%) after surgery. Urinary extravasation at 4 days after surgery was followed by same rate of BN sclerosis and urinary continence as in patients with no urinary extravasation. CONCLUSION: BN mucosal eversion before vesico-urethral anastomosis during RRP is not essential to reduce the frequency of BN sclerosis or urinary incontinence. Early radiological urinary extravasation at the vesico-urethral anastomosis did not increase the risk of BN sclerosis or urinary incontinence. PMID- 15794778 TI - Prostate-specific antigen (PSA) complexed to alpha1-antichymotrypsin improves prostate cancer detection using total PSA in Japanese patients with total PSA levels of 2.0-4.0 ng/mL. AB - OBJECTIVE: To assess the utility of prostate-specific antigen (PSA) complexed to alpha1-antichymotrypsin (PSA-ACT) in prostate cancer screening in Japanese men with a total PSA level of 2.0-4.0 ng/mL, as improving cancer detection in men with these total PSA levels is a challenge for clinical urologists. PATIENTS AND METHODS: Total PSA and PSA-ACT were prospectively assessed and prostate biopsy recommended for patients who met either of two thresholds, i.e. a total PSA of > or = 2.0 ng/mL or a PSA-ACT of > or= 1.5 ng/mL. The diagnostic ability of total PSA and PSA-ACT, and free-to-total PSA ratio and prostate volume-adjusted density were evaluated by receiver operating characteristic (ROC) analysis. RESULTS: Of 1003 men enrolled, 547 met the biopsy criteria and a biopsy was taken in 315 (57.6%) patients. The area under the ROC curve for PSA-ACT (0.679) was significantly greater than that for total PSA (0.601, P = 0.04) and equivalent to that for the free-to-total ratio (0.686, P = 0.911) in 116 men, including 27 with cancer with total PSA levels of 2.0-4.0 ng/mL. PSA-ACT was more specific than the free-to-total ratio at a sensitivity of 95% (36% vs 18%, P < 0.05). The best variable for discriminating between cancer and benign disease in men with PSA levels of 2.0-4.0 ng/mL was PSA-ACT density (area under the curve 0.852) which provided 66% specificity at a sensitivity of 90%. CONCLUSIONS: PSA-ACT is better than total PSA and equivalent to the free-to-total ratio for detecting prostate cancer in men with PSA levels of 2.0-4.0 ng/mL, and is thus useful for reducing the number of unnecessary biopsies. PMID- 15794779 TI - A novel technique for approaching the endopelvic fascia in retropubic radical prostatectomy, based on an anatomical study of fixed and fresh cadavers. AB - OBJECTIVE: To present the anatomical basis and details of a technique for an approach to the endopelvic fascia devised to preserve urinary continence. PATIENTS, MATERIALS AND METHODS: For cross-sectional macroscopic observation, seven formalin-fixed specimens of the male pelvic contents including the pelvic wall were serially sectioned at a 5-mm thickness. Semi-serial sections from eight other specimens were examined histologically. Eight fresh cadavers were dissected to mimic the various steps in a retropubic radical prostatectomy. After approaching the endopelvic fascia in an anatomically determined manner to reach the paraprostatic space, the pubic bone was removed and nerves near the rhabdosphincter dissected. To assess the clinical implication of this approach, we examined the time to achieve continence in 23 consecutive patients who had a radical retropubic prostatectomy using the new technique. RESULTS: Sectional macroscopic observation depicted the fascia of the levator ani as a definite structure adherent to but not fused with the lateral pelvic fascia. The thin fascia overlying the levator ani fascia and lateral pelvic fascia represented the true endopelvic fascia. Microscopically, the lower part of the fascia of the levator ani was rich in smooth muscle, which interdigitated with the framework of the rhabdosphincter. In fresh cadavers, the levator ani muscle was removed laterally still covered by its fascia, without visualizing the muscle fibres. Small branches from the pudendal nerve entered the rhabdosphincter. The mean (sd, range) distance from the lowest point of the endopelvic fascia to the point where the sphincteric branch entered the rhabdosphincter was 5.5 (1.8, 3-8) mm. The continence rate at 1, 3, 6 and 9 months after surgery using the new technique was 44%, 83%, 96% and 100%, respectively. CONCLUSIONS: Preserving the fascia of the levator ani helps to protect the levator ani muscle, rhabdosphincter and pudendal nerve branches to the rhabdosphincter. In retropubic radical prostatectomy, this anatomical approach to the endopelvic fascia should preserve or allow the earlier recovery of urinary continence. PMID- 15794780 TI - The incidence and treatment of lymphoceles after radical retropubic prostatectomy. AB - OBJECTIVE: To determine the incidence and treatment of lymphoceles after retropubic radical prostatectomy (RP). PATIENTS AND METHODS: Up to January 2004, 260 patients who had a retropubic RP in one institution by one surgeon were assessed retrospectively, using the patients' notes or the computerized results system to determine whether a lymphocele was suspected and then confirmed by imaging studies (computed tomography or ultrasonography). RESULTS: Nine patients developed symptomatic lymphoceles; eight of these were detected by imaging. Four lymphoceles required intervention while the remainder regressed spontaneously. No complications were reported in the group that was treated. CONCLUSION: The rate of symptomatic lymphocele formation was low after RP, with an overall incidence of 3.5%. Ultrasonography was effective in detecting lymphoceles and ultrasonographically guided percutaneous drainage an effective treatment. PMID- 15794781 TI - Testosterone recovery and changes in bone mineral density after stopping long term luteinizing hormone-releasing hormone analogue therapy in osteoporotic patients with prostate cancer. AB - OBJECTIVE: To investigate the rate of testosterone recovery and changes in bone mineral density in patients found to be osteoporotic while receiving luteinizing hormone-releasing hormone (LHRH) analogues after changing to antiandrogen monotherapy in an attempt to reduce further demineralization. PATIENTS AND METHODS: Fifteen patients receiving LHRH analogue therapy for > or = 1 year were identified as osteoporotic by distal forearm dual X-ray densitometry. They were then converted to antiandrogen monotherapy, and prostate specific-antigen (PSA) and total testosterone monitored at 3-monthly intervals. The forearm densitometry was repeated at 1 year. RESULTS: All patients had some testosterone recovery; the mean (range) duration to initial detectable testosterone was 12.8 (6-22) months. Six patients had a normal testosterone level after a mean of 17.5 (14-30) months. In the year after stopping LHRH analogue therapy the mean bone mineral density (t score) decreased by 7.2%. CONCLUSIONS: Osteoporotic patients, after stopping LHRH analogues, continue to have suppressed levels of testosterone which have a detrimental effect on bone mineral density. We therefore would not advocate conversion to antiandrogen monotherapy to improve bone density, and suggest alternative therapeutic intervention e.g. bisphosphonate therapy, for these patients. PMID- 15794782 TI - Sexual, psychological and dyadic qualities of the prostate cancer 'couple'. AB - OBJECTIVES: To examine the levels of sexual, psychological and dyadic functioning of the prostate cancer 'couple' (as studies have shown that spouses/partners play an integral role in the patient's adjustment to prostate cancer treatment), to encourage the creation of innovative psychosexual interventions to be used in the outpatient setting, and to offer insights into a novel area of prostate cancer research. PATIENTS AND METHODS: In all, 103 men newly diagnosed with prostate cancer, and their partners, were assessed in an academic outpatient setting using instruments measuring sexual function, depressed mood, psychological distress and dyadic adjustment. RESULTS: The partners' mean scores on sexual function questions were 55.75, significantly higher than those of the patients (51.7, P = 0.018), showing that partners perceived their sexual performance at a better level. Partners' mean scores on the depression and distress measures were also significantly higher. On those items that monitored the accuracy of the patients' perceptions of their sexual function, partners rated the patients significantly lower in ability to gain erections (patient/partner means 2.67/4.52; P < 0.001) and to perform sexually (patient/partner means 1.38/4.68; P < 0.001) than they rated themselves. CONCLUSIONS: Information from this study could be useful in constructing interventions that allow the physician and the prostate cancer 'couple' to reflect on issues of sexual function and psychological distress that might once have been considered taboo. The results characterize the disparities between patients with prostate cancer and their partners on self-reported questionnaires, and underscore how important it is to hear the voice of the 'couple'. PMID- 15794783 TI - Correlation between clinical and pathological staging in a series of radical cystectomies for bladder carcinoma. AB - OBJECTIVE: To analyse the rate of concordance between the clinical and pathological Tumour-Nodes-Metastasis staging systems in a homogeneous series of patients who had undergone radical cystectomy for locally advanced or recurrent multifocal superficial bladder carcinoma. PATIENTS AND METHODS: The clinical data of 156 patients who had undergone radical cystectomy and bilateral iliaco obturator lymphadenectomy for bladder cancer in our department were analysed retrospectively. RESULTS The clinical stage of the primary tumour was carcinoma in situ in three patients (1.9%), cT1 in 67 (42.9%), cT2 in 70 (44.9%), cT3 in five (3.2%) and cT4 in nine (5.8%). Clinical lymph node involvement was detected in 19 patients (12.2%). The differences between clinical and pathological stages were statistically significant (P < 0.001), the concordance was moderate (kappa = 0.27, P < 0.001). Of the 70 patients with < or = cT1, 40 (57%) were reconfirmed as having pathological stage < or = T1; of the 70 with cT2, 16 (23%) had pT2 carcinoma. Of the 140 patients with clinically organ-confined (< or =T2) neoplasms, 70 (50%) had been understaged after radical cystectomy. The clinical and pathological systems were statistically overlapping for locally advanced cases only. Pathological lymph node involvement was diagnosed in 45 patients (28.8%); this was foreseen with pelvic computed tomography in 19 (12%) only (P < 0.001). All patients designated cN+ were also pN+. CONCLUSION: These data confirm the high risk of clinical understaging of both local extension of the primary tumour and lymph node involvement. PMID- 15794784 TI - A comparison of the pathology of transitional cell carcinoma of the bladder and upper urinary tract. AB - OBJECTIVE: To clarify the histopathological patterns of upper and lower urinary tract transitional cell carcinomas (TCCs), as previous reports suggest that upper urinary tract TCCs have a greater tendency towards high-grade disease than bladder TCCs, of which most are low-grade and low-stage tumours. PATIENTS AND METHODS: All patients presenting with TCC of bladder or upper urinary tract between February 1991 and December 2001 at one institution were identified. Further patient information was obtained from the hospital database and case-note review. RESULTS: In all, 164 patients with upper urinary tract TCC and 2197 with bladder TCC were identified. There was a correlation between grade and stage of both upper urinary tract and bladder TCCs. 35% of the upper tract TCCs were classified as grade 2 and 44% as grade 3, while for bladder TCCs, 31% of lesions were classified as grade 2 and 35% as grade 3 (P = 0.003). Of the upper urinary tract lesions 33% were stage pT2-T4, compared with only 20% of bladder TCCs (P = 0.001). CONCLUSIONS: Upper urinary tract TCC is a higher grade and stage disease than bladder cancer, a finding that emphasizes the need for aggressive treatment of upper urinary tract TCC. If endourological management of upper urinary tract TCC is considered, histopathological determination of tumour grade before treatment is essential. PMID- 15794785 TI - The assessment of patient life-expectancy: how accurate are urologists and oncologists? AB - OBJECTIVE: To assess the degree of accuracy, precision and consistency with which consultant urologists, oncologists and junior doctors predict a patient's 10-year life-expectancy. SUBJECTS AND METHODS: Eighteen doctors of varying seniority independently examined 70 patient case scenarios containing detailed medical histories; 13 of these cases were duplicate scenarios. Bland-Altman analyses were used to compare doctors' estimates of the probability of each hypothetical patient surviving 10 years with that calculated using actuarial methods. Intra- and interdoctor reliability were also assessed. RESULTS: Compared with actuarial estimates, doctors underestimated the 10-year survival probability by an overall mean of 10.8% (95% confidence interval, 10.1-11.5%). The 18 individual doctors ranged from a mean underestimation of 33.2% to a mean overestimation of 3.9%. Variation around these means was considerable for each doctor, the standard deviations being 14.5-20.9%. Inter-doctor reliability was 0.58, while overall intra-doctor reliability was 0.74, but for individual doctors was 0.31-0.94. Junior doctors were less accurate in their predictions than the senior doctors. Five doctors tended to overestimate where life-expectancy was poor and underestimate where it was good. CONCLUSIONS: Doctors were poor at predicting 10 year survival, tending to underestimate when compared with actuarial estimates. There was also substantial variability both within and between doctors. The inaccuracy, imprecision and inconsistency amongst the doctors in assessing patient life-expectancy is an important finding and has significant implications for managing patients. Many patients may be denied treatment after a pessimistic assessment of life-expectancy and (less commonly) some may inappropriately be offered treatment after an optimistic assessment. The particular inaccuracy in junior doctors compared with their senior colleagues also highlights the need for training. The development of a tool to assist in both training and clinical practice has the potential to improve doctors' decision-making and patient care. PMID- 15794786 TI - Towards a better understanding of involuntary detrusor activity. AB - There is a wide spectrum of topics covered in this section. The ever-present problem of nocturia is further explored by authors from Sweden, with interesting conclusions. In addition, several papers describe various issues relating to the surgical correction of stress urinary incontinence. Finally, authors from Switzerland describe their use of sacral magnetic stimulation in non-inflammatory chronic pelvic pain syndrome. OBJECTIVES: To compare the measured variables of involuntary detrusor activity (IDA) during ambulatory cystometry (AC) in women with and with no overactive bladder symptoms, and to evaluate the correlation between these variables and symptom severity. PATIENTS AND METHODS: In all, 61 symptomatic and 39 asymptomatic women completed the Bristol Female Lower Urinary Tract Symptoms Questionnaire (BFLUTS-Q) and underwent AC. Measured variables of IDA (amplitude, duration, bladder volume and symptoms) were compared in the two cohorts. RESULTS: IDA was detected in 47 of 61 symptomatic women (77%) and in 17 of 36 (47%) controls (P < or = 0.01). The maximum IDA (defined as the highest amplitude contraction in any fill-void cycle) occurred at significantly lower volumes (328 vs 450 mL, P < or = 0.05), was of higher amplitude (26 vs 12 cm H(2)O, P = 0.14) and longer duration (83 vs 14 s, P < or = 0.05) in symptomatic women than in controls. There was coincident incontinence in 22 (36%) symptomatic women and no controls (P < or = 0.01). Discriminatory levels for clinically relevant IDA were established, and when applied retrospectively, classified 35 of 61 symptomatic women (55%) and one control (3%) as having such IDA. The correlation between symptom severity (measured by the BFLUTS-Q) and urodynamics was higher when these criteria were applied (r = 0.52 vs 0.38, P < or = 0.01). CONCLUSION: There are quantifiable differences between IDA found during AC in symptomatic and asymptomatic women. The measured variables of IDA may be useful to determine its clinical relevance, which may be indicated by contractions associated with leakage or contractions of >30 s occurring at bladder volumes of <300 mL. PMID- 15794788 TI - Self-assessed health, sadness and happiness in relation to the total burden of symptoms from the lower urinary tract. AB - OBJECTIVES: To evaluate the effect of lower urinary tract symptoms (LUTS) on self assessed health, sadness and happiness of men. SUBJECTS AND METHODS: The study included 504 men (aged 40-80 years) in the rural community of Surahammar, Sweden, who a year earlier had reported stress incontinence, urgency or postvoid dribbling in answer to a postal questionnaire, and 504 age-matched control men from the same community. The occurrence of 12 specific LUTS was rated using the Danish Prostatic Symptom Score. Health, sadness and happiness were measured by three questions from the Medical Outcomes Study Short-Form 36 health survey questionnaire. RESULTS: Completed questionnaires were returned by 74.2% of men (748/1008). A low score for health was reported by 34% of men with one to four LUTS, by 67% with five to eight, and by 75% with nine or more LUTS. The total LUTS burden correlated with lower scores for happiness and with higher scores for sadness. For each of the 12 specific LUTS, men with the symptom had lower scores for health and happiness, and higher scores for sadness, than men without the symptom. Comparing men with the symptom of 'other incontinence' to men with no 'other incontinence', the relative risk (95% confidence interval) of impaired health was 2.2 (1.8-2.8), while that of a high score for happiness was 0.5 (0.3 0.7) and that of greater sadness was 2.3 (1.7-3.3). Social status, marital status, education, smoking, physical activity and urinary tract infection all affected the impact of LUTS. CONCLUSIONS: The total burden of LUTS is related to self-assessed health, sadness and happiness. PMID- 15794787 TI - The pharmacokinetics of 400 microg of oral desmopressin in elderly patients with nocturia, and the correlation between the absorption of desmopressin and clinical effect. AB - OBJECTIVE: To investigate the pharmacokinetic profile of oral desmopressin in elderly patients with nocturia, and to analyse any possible correlation between the absorption and clinical effect. PATIENTS AND METHODS: In all, 32 patients were screened to determine the baseline number of nocturnal voids and the nocturia index; of these, 24 fulfilled the inclusion criteria and were enrolled for a pharmacokinetic evaluation of oral desmopressin 400 microg. A double-blind, randomized, placebo-controlled, crossover-effect evaluation period was then used to test the association between the absorption of desmopressin and pharmacodynamic effect. Serial plasma samples were collected for 8 h for a pharmacokinetic analysis of desmopressin. The pharmacodynamics after an equivalent oral dose before bedtime were assessed by measuring changes in the number of nocturnal voids, time to first nocturnal void and nocturnal diuresis, from placebo to active treatment. RESULTS: There was a linear relationship between plasma desmopressin at 2 h after dosing and the area under the plasma concentration curve from 0 to infinity (Pearson's rho 0.923, P < 0.001). Women had a significantly higher plasma desmopressin concentration than men (P = 0.0012) and more adverse events. There was no correlation between plasma desmopressin at 2 h after dosing and the within-patient response in any of the effect variables. Generally, the number of nocturnal voids and nocturnal diuresis were half that with placebo. The time to the first nocturnal void was almost doubled compared with placebo. CONCLUSIONS: There seems to be a relationship between gender, plasma level of desmopressin and the incidence of adverse events. Plasma desmopressin at 2 h after dosing cannot be used to predict the pharmacodynamic response, although desmopressin lowers the nocturnal diuresis and the number of nocturnal voids. PMID- 15794789 TI - Nocturia in relation to somatic health, mental health and pain in adult men and women. AB - OBJECTIVE: To assess the relationship of nocturia to somatic health, mental health and bodily pain. SUBJECTS AND METHODS: A randomly selected group of men and women aged 20-64 years, living in three small municipalities in northern Sweden, or in the city of Ostersund or in Stockholm, were sent a postal questionnaire containing questions on somatic and mental health, satisfaction with life, pain, nocturnal voiding, work and sick-listing from work. RESULTS: Reports (from 1948 respondents) on poor somatic and mental health and on pain all increased in parallel with increasing frequency of nocturnal voids. In a multiple logistic regression analysis with sex, age, somatic health, mental health and bodily pain as the independent variables, significant independent correlates (odds ratios, confidence intervals) of nocturnal micturition (two or more episodes vs none or one) were: age 45-59 vs 20-44 years, 1.9 (1.3-2.7), > or =60 vs 20-44 years, 3.8 (2.4-6.0); somatic health, poor vs good, 2.3 (1.4-3.7); mental health, poor vs good, 1.9 (1.2-3.0); pain, rather mild vs very mild or none, 1.5 (1.0-2.3); rather severe vs very mild or none, 1.9 (1.1-3.2); and very severe vs very mild or none, 6.0 (2.5-14.0). Gender was deleted by the logistic model. Sick-listing for > or = 60 days during the past year was reported by 4.9%, 10.6%, 5.6% and 38.9% of the men with none, one, two or > or = three nocturnal voids, respectively, and by 10%, 12.4%, 23% and 46.7% (both P < 0.001) of the corresponding women, respectively. Life satisfaction decreased in parallel with increased nocturia. CONCLUSION: The impairment of both somatic and mental health was associated with increased nocturnal voiding. Pain was associated with a substantial increase in nocturia after adjusting for age and somatic and mental health. Sick-leave was more common in association with more nocturnal voids. PMID- 15794790 TI - Nocturia, depression and antidepressant medication. AB - OBJECTIVE: To assess the possible relationship between treatment with selective serotonin-reuptake inhibitors (SSRIs) and the occurrence of nocturia. SUBJECTS AND METHODS: An unselected group of adult men and women, living in the city of Ostersund, Sweden, were sent a postal questionnaire containing questions on somatic and mental health, sleep, sleepiness and nocturia. For depression diagnostics, the Major Depression Inventory (MDI) was used. Prescription data on antidepressant drugs were extracted from a register in the county of Jamtland, Sweden. RESULTS: The questionnaire was completed by 1375 subjects, of whom 609 (44%) were men; the response rate was 69%. Prescription data were available for all respondents. The mean (sd) age of the men and women participating were 48.0 (18.2) and 50.1 (19.1) years, respectively. Two or more nocturnal micturition episodes were reported in 15.6% of the men and 16.5% of the women. In a multiple logistic regression analysis, independent correlates for two or more nocturnal voids vs no more than one were: age 45-59 years vs <45 (odds ratio 2.9; 95% confidence interval 1.9-4.7); age 60-74 vs <45 (6.0; 3.7-9.8); age > 75 vs <45 (13.4; 7.9-22.6); major depression, yes vs no (4.6; 2.8-7.5); and being on treatment with SSRI, yes vs no (2.2; 1.1-4.5). Gender was deleted by the logistic regression model. CONCLUSION: Major depression has previously been found to be associated with increased nocturnal micturition. In the present study, twice as many men and women treated with SSRIs as those not so treated had two or more nocturnal voids, after adjusting for major depression and age. The implication for the risk of fall injuries is discussed. PMID- 15794792 TI - Day-case sling surgery for stress urinary incontinence: feasibility and safety. AB - OBJECTIVE: To prospectively assess the feasibility for discharge 10 h after a porcine dermal pubovaginal sling procedure (PVS), to examine the surgical factors (postoperative complications) affecting discharge, and to measure the short-term cure rate for stress urinary incontinence (SUI). PATIENTS AND METHODS: Between June 2003 and December 2003, 40 consecutive patients with SUI and scheduled for treatment using a porcine dermal sling were enrolled in this prospective study. Patients were admitted with a planned overnight stay and returned to the ward with no urinary catheter. Outcome measures were bladder emptying efficiency (EE) at 10 h after surgery, time intervals to the first three spontaneous voids, EE of the first three voids, time required to achieve an EE of > or = 75%, a visual analogue scale pain score, perioperative complications, and short-term cure rate of SUI. Patients were considered suitable for discharge from hospital when the EE was > or = 75% or when they were self-catheterizing confidently with adequate pain control and no significant complication. All patients were followed for 6 months. RESULTS: The median EE at 10 h was 61%; 16 patients (40%) achieved efficient emptying and were suitable for discharge 10 h after surgery. The median intervals to the first three spontaneous voids were 7, 10 and 17 h, and the median EEs for the first three voids 46%, 61% and 75%. The median visual analogue scale pain score was 3.5. Patients with intrinsic sphincter deficiency (ISD) were significantly less likely to achieve efficient emptying at 10 h (39% vs 70%). Overall SUI was cured or improved in 90% of patients at the 6-month follow-up. CONCLUSIONS: In the present study only 40% of patients were suitable for day-case sling surgery. Early bladder emptying inefficiency was the main limiting factor. Exclusion of patients with ISD and possibly decreasing the EE threshold to 50% would improve the discharge rate. The short-term results of this PVS are similar to those obtained with the autologous fascial sling. PMID- 15794791 TI - Combined external urethral bulking and artificial urinary sphincter for urethral atrophy and stress urinary incontinence. AB - OBJECTIVE: To describe a technique of externally bulking the urethra with a soft tissue graft before placing another artificial urinary sphincter (AUS), as when placing another AUS for recurrent male stress urinary incontinence (SUI) other manoeuvres, e.g. placing a tandem cuff or transcorporal cuff, must be used to obtain urinary continence in an atrophic urethra, and each is associated with morbidity. PATIENTS AND METHODS: From January 2003 to July 2004, five patients (mean age 74 years, range 62-84) treated by radical prostatectomy were referred for recurrent SUI after placing an AUS (four, including one with urethral erosion) or a male sling (one, with a resulting atrophic urethra). Each patient was treated with an external urethral bulking agent (Surgisis) ES, Cook Urological, Spencer, Indiana) and had an AUS placed. RESULTS: In each patient the greatest urethral circumference was <4 cm. To place a functional 4 cm cuff, the diameter of the urethra was enhanced by wrapping it with Surgisis ES. Continence was significantly improved in all patients except one 84-year-old man who had the replanted artificial sphincter removed because of erosion 14 months after surgery. CONCLUSION: In cases of severe recurrent SUI from urethral atrophy after placing an AUS, externally bulking the urethra with Surgisis ES before placing another AUS is well tolerated, and gives satisfactory results. PMID- 15794794 TI - Sacral magnetic stimulation in non-inflammatory chronic pelvic pain syndrome. AB - OBJECTIVES: To prospectively evaluate sacral magnetic high-frequency stimulation as a treatment option for patients with non-inflammatory chronic pelvic pain syndrome (CPPS, category IIIB). PATIENTS AND METHODS: Fourteen men with CPPS IIIB were treated with high-frequency sacral magnetic stimulation, with 10 treatment sessions once a week for 30 min at a frequency of 50 Hz. The National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) and quality-of-life index were determined before and after treatment. RESULTS: All patients tolerated the stimulation well and 12 of 14 reported agreeable sensations during stimulation. There were no complications; only one patient did not complete the treatment course. The mean (range) total NIH-CPSI score did not change with treatment, at 27 (18-38) before and 27 (4-40) after treatment. Moreover, there was no sustained effect on the mean scores for pain, micturition complaints or quality of life. CONCLUSIONS: High-frequency sacral magnetic stimulation in patients with CPPS IIIB only reduces pain during stimulation, with no sustained relief of symptoms. Therefore, intermittent sacral magnetic stimulation cannot be recommended as a treatment option for CPPS IIIB. PMID- 15794793 TI - A stereological analysis of fibrosis and inflammatory reaction induced by four different synthetic slings. AB - OBJECTIVES: To analyse quantitatively, using stereological methods, the density of the collagen fibres induced by four types of sling materials, and verify by a histopathological analysis the corresponding inflammatory reaction, as fibrosis secondary to sling implantation is considered responsible for restoring urethral support and re-establishing continence in women with stress urinary incontinence, and new synthetic materials that promote adequate fibrosis with the least intensity and duration have been proposed to substitute the aponeurotic sling. MATERIALS AND METHODS: The study comprised 70 isogenic white Wistar rats divided into three groups: group A (30 rats) had 8 x 4 mm strips of silicone and porcine small intestine submucosa (SIS) implanted in the abdominal subcutaneous tissues; group B (30 rats) had 8 x 4 mm strips of polycaprolactone and polylactic acid copolymers and monofilament polypropylene (PLP) implanted the abdominal subcutaneous layer; while a control group of 10 rats had dissection and suturing with 5/0 Nylon in the abdominal subcutaneous layer, as used to fix the strips in the other rats. Picro-Sirius staining was used to assess collagen fibres, and haematoxylin-eosin for the histopathological study. At 7, 30 and 90 days after surgery, 10 rats from each group were killed and assessed. RESULTS: After 7 days all the materials induced a moderate inflammatory reaction that did not differ from that in the control group. At 30 days there was no difference between the control and polycaprolactone and polylactic acid copolymers, having the least inflammatory reaction. PLP and silicone produced a moderate inflammatory reaction, while the porcine SIS induced a more intense reaction. At 90 days there was a more intense inflammatory reaction in polycaprolactone and polylactic acid copolymers than before but it was less than with PLP and silicone, which again were no different. During this period the inflammatory reaction induced by SIS was greater. The stereological analysis indicated that collagen fibres induced by polycaprolactone and polylactic acid copolymers and PLP were less dense (61% and 65%, respectively), and significantly less than with silicone (85%) and SIS (86%). CONCLUSION: PLP was the best nonabsorbable material as it induced a less intense inflammatory reaction than the other tested materials. As porcine SIS was completely absorbed the intense fibrosis induced is useful, as it is exclusively responsible for the urethral support later after surgery. PMID- 15794795 TI - Use of combined intracorporal injection and a phosphodiesterase-5 inhibitor therapy for men with a suboptimal response to sildenafil and/or vardenafil monotherapy after radical retropubic prostatectomy. AB - OBJECTIVE: To report experience with combined therapy using intracorporal injection (ICI) of alprostadil and oral phosphodiesterase 5 (PDE-5) inhibitors for the minimally invasive treatment of erectile dysfunction (ED) after radical prostatectomy (RP), as PDE-5 inhibitors are effective but a few patients may have a suboptimal response. PATIENTS AND METHODS: In a retrospective study, 34 men (aged 46-66 years) had a nerve-sparing retropubic RP and subsequent ED. Patients were titrated on sildenafil citrate or vardenafil to maximum doses. All had a suboptimal response after a maximum of eight doses of oral therapy and were then treated with ICI therapy using 15 or 20 microg alprostadil. Erectile function was assessed with the Sexual Health Inventory for Men (SHIM). RESULTS: Of the 32 patients who continued combined therapy, 22 (68%) had an improvement in erectile function after ICI therapy, as assessed by the SHIM score. On follow-up, 36% of these patients used ICI therapy only intermittently, instead of regularly, as they felt that this was adequate enough for good results. CONCLUSIONS: PDE-5 oral pharmacotherapy is the most commonly used effective therapy for ED but may not be as effective in patients who have radical surgery; the addition of testosterone patches may have side-effects or be considered a risk in patients with a history of prostate cancer. The use of ICI therapy as an adjunct or maintenance therapy to their oral medication may be another alternative in these patients. PMID- 15794796 TI - The effect on erectile function of 103palladium implantation for localized prostate cancer. AB - OBJECTIVE: To determine in a prospective study the effect on erectile function of (103)Pd brachytherapy for localized prostate cancer, using a validated questionnaire. PATIENTS AND METHODS: Between July 1999 and April 2003, 113 men with localized prostate cancer were treated by permanent implantation of (103)Pd seeds, of whom 78 with a follow-up of 30 months were included in this study. No patient received supplemental external beam radiation therapy. At baseline and 3 month intervals, erectile function (EF) was assessed by the EF domain score of the International Index of Erectile Function-15 (IIEF-15); 77% received (neo)adjuvant antiandrogen therapy for up to 3 months. RESULTS: At baseline, 27 (35%) patients had no erectile dysfunction (ED; EF domain score 26-30), 24 (31%) had mild/moderate ED (score 11-25) and 27 (35%) severe ED (score 6-10). The mean EF domain score decreased from 17 to 12 (P < 0.001) after 30 months. Overall, 52 men (67%, including those with severe ED at baseline) remained in the same ED category at 30 months after therapy as before, 12 (15%) deteriorated by one category, 14 (18%) by two or more, and no patient improved. Of the 27 patients fully potent (score 26-30) at baseline, 37% remained so after 30 months, 19% developed mild and the remaining 44% moderate/severe ED. In a multivariate analysis, neither age nor preoperative prostate-specific antigen level, prostate volume, D90, hormonal treatment, diabetes, smoking or hypertension were predictive of preserving potency (P > 0.05). CONCLUSIONS: There was a high prevalence of pre-existing ED in these men; 57% of men fully potent or with mild ED at baseline remained so 30 months after brachytherapy. PMID- 15794797 TI - Comparison of laparoscopic and open donor nephrectomy: a randomized controlled trial. AB - Authors from Iran compare various outcomes between laparoscopic and open donor nephrectomy in kidney transplantation; they carried out a large comparative trial, and found that laparoscopic donor nephrectomy gave better donor satisfaction and morbidity, with equivalent graft outcome. OBJECTIVE: To compare the graft survival, donor and recipient outcome, donor satisfaction, and complications of laparoscopic (LDN) and open donor nephrectomy (ODN) in kidney transplantation. PATIENTS AND METHODS: In a randomized controlled trial, 100 cases each of LDN and ODN were compared. We modified the standard LDN procedure to make it less expensive. RESULTS: The mean (sd) operative duration was 152.2 (33.9) min for ODN and 270.8 (58.5) min for LDN, and the mean duration of kidney warm ischaemia was 1.87 min for ODN and 8.7 min for LDN. Only one LDN required conversion to ODN because of bleeding. The mean follow-up in the LDN and ODN groups was not significantly different (406.1 vs 403.8 days). The mean (sd) score for donor satisfaction was 17.3 (3.5) for ODN and 19.6 (1.0) for LDN. The rate of ureteric complications was 2% for ODN and none for LDN. As determined by serum creatinine levels at 3, 21-30, 90, 180 and 365 days after surgery, graft function was not significantly different between ODN and LDN. Long-term graft survival was 93.8% for LDN and 92.7% for ODN. CONCLUSIONS: Compared to ODN, LDN was associated with greater donor satisfaction, less morbidity and equivalent graft outcome. PMID- 15794798 TI - Determination of the time required for appropriate chemical de-epithelialization of an ileal segment for cystoplasty: an animal model. AB - Another group of authors from Iran attempted experimentally to determine the required time for the appropriate enzymatic treatment of the ideal segment to complete de-epithelialization, thus reducing its absorptive function. They found that 25 min of enzymatic treatment of the ideal segment was adequate for this, and that it was recommended from their experimental study for cystoplasty. OBJECTIVES: To determine the time required for the appropriate enzymatic treatment of an ileal segment to de-epithelialize its mucosa and to reduce its absorptive function for cystoplasty in dogs. MATERIALS AND METHODS: Twenty-one adult female Persian mixed-breed dogs were divided into seven equal groups: group 1 (negative control group) had no ileocystoplasty; group 2 (positive control group) had a routine ileocystoplasty with no enzymatic treatment of the ileal segment; and groups 3-7 had an ileocystoplasty with 5, 10, 15, 20 or 25 min, respectively, of enzymatic treatment of the ileal segment with collagenase and trypsin. The seven groups were then compared for haematological, biochemical and histological changes, and glucose reabsorption assessed using a glucose absorption test. RESULTS: No dogs showed any signs of metabolic disturbances, biochemical and haematological changes. There were significant differences in blood glucose level (BGL) for the groups at different times after the glucose absorption test, but a pair-wise comparison showed significant differences in BGL between group 1 and the other groups, except group 7. The degree of histopathological change was associated with the duration of enzymatic treatment, in that changes were more prominent in group 7. There was no shrinkage or collagen deposition. CONCLUSIONS: In these conditions, 25 min of enzymatic treatment of the ileal segment is sufficient to remove the absorptive function of the augmented bladder, and is recommended for cystoplasty in dogs. PMID- 15794799 TI - The laparoscopic management of intersex patients: the preferred approach. AB - In the world's largest series of patients with intersex treated by laparoscopy, authors from Sao Paulo found that this technique allowed easy identification and removal of gonads. They also found that other organs could be removed and genitoplasty performed. OBJECTIVE: To present possibly the largest series of the use of laparoscopy for treating intersex patients. PATIENTS AND METHODS: Fifty intersex patients (34 with male and two with female pseudohermaphroditism, nine with gonadal dysgenesis, four with true hermaphroditism, and one with complex hypospadias), aged 0.5-46 years (mean 18.3), underwent laparoscopy to remove gonads and/or ductal structures incompatible with the social gender, or for gonadal tumour or a potential risk for malignancy. When necessary, genitoplasty was performed concomitantly. RESULTS: At the laparoscopic evaluation, 10 gonads of six patients were absent, while four were identified as 'vanishing'; 72 gonads (46 dysgenetic, 17 normal testes, one normal ovary, one ovotestis, seven gonadoblastomas or dysgerminomas) were removed; two ovotestes were replaced in the scrotum after removing the ovarian segment, as was one normal testis. Twelve patients with a urogenital sinus had its vaginal component removed, 11 including a hysterectomy. Three of these patients had a combined perineal approach to complete its removal, together with masculinizing genitoplasty. There were no intraoperative complications or conversions; two patients had complications after surgery. CONCLUSIONS: Laparoscopy allows the straightforward identification and removal of gonads. All abnormal ductal structures must be removed, as this increases the chance of resecting unidentified gonads. Removing the uterus and vaginal component of the urogenital sinus in patients with male social sex is feasible, with low morbidity. Genitoplasty, according to the social sex, can be performed in the same procedure. PMID- 15794801 TI - The promise of gene-expression analysis in bladder cancer: a clinician's guide. PMID- 15794802 TI - Synergistic inhibitory effect of high-intensity focused ultrasound combined with chemotherapy on Dunning adenocarcinoma. AB - OBJECTIVE: To evaluate the therapeutic effect of high-intensity focused ultrasound (HIFU) combined with chemotherapy (paclitaxel + estramustine) on AT2 Dunning adenocarcinoma, as no satisfactory treatment for localized prostate cancer is available for patients with a poor prognosis, e.g. stage T3, a high Gleason score, or a prostate-specific antigen level of >15 ng/mL. MATERIALS AND METHODS: Forty-one Dunning AT2 tumour-bearing Copenhagen rats were divided into four groups, i.e. control, chemotherapy, HIFU, and chemotherapy + HIFU (the last three treated for 1 week). The growth in tumour volume was recorded for 3 weeks, the point at which tumour volume was considered to have doubled (doubling time). The growth curves of each group were plotted and evaluated statistically. RESULTS: At 30 days of follow-up the distributions of tumour volume with treatment group were significantly different (P < 0.001); volumes were significantly greater in the control than in the chemotherapy-only or in the HIFU only group (both P = 0.006). The greatest difference was between the chemotherapy + HIFU and the control group. The tumour doubling times were 13.2 days for HIFU only, 31.2 days for chemotherapy + HIFU and 7.7 days for the controls. CONCLUSION: These results suggest that this combined therapy could be useful for treating patients with high-risk prostate cancer. PMID- 15794800 TI - Predictive factors of ultrasonographic involution of prenatally detected multicystic dysplastic kidney. AB - OBJECTIVE: To evaluate possible predictive factors of involution on ultrasonography (US) or disappearance of a prenatally detected multicystic dysplastic kidney (MCDK). PATIENTS AND METHODS: Forty-five children with unilateral MCDK detected by prenatal ultrasonography between 1989 and 2002 were analysed. All patients except one had (99m)Tc isotopic scintigraphy to confirm the absence of renal function in the MCDK. All children were managed conservatively with follow-up visits every 6 months, with US 6-monthly during the first 2 years of life and yearly thereafter. Survival was analysed using the Kaplan-Meier method to evaluate the involution of the MCDK, with differences between subgroups assessed using the two-sided log-rank test. Cox's regression model was applied for the multivariate analysis. RESULTS: The mean (range) follow up was 50 (12-167) months; in all, 279 ultrasonograms were taken, the mean (range) number per patient being 6 (3-10). US showed partial involution of the MCDK in 30 (67%) cases and complete involution in nine (20%). The absolute MCDK length remained almost unchanged in six children (13%). Univariate analysis showed that four variables were possibly associated with complete involution of the MCDK (gender, impalpable kidney, renal length at admission using two thresholds, 62 and 78 mm). After adjusting by Cox's model only a renal length at diagnosis of <62 mm remained associated with complete involution (relative risk 8, 95% confidence interval 0.98-68; P = 0.05). CONCLUSION: These results suggest that only a renal length of <62 mm on initial US was predictive of complete involution of the MCDK during the follow-up. PMID- 15794803 TI - Stereotactic electrical stimulation of the pontine micturition centre in the pig. AB - OBJECTIVE: To apply stereotactic electrical stimulation of the pig brainstem and thus identify a pontine micturition centre. MATERIALS AND METHODS: In 10 anaesthetized female Vietnamese minipigs a needle-electrode was positioned in the pontine region. Pressure responses in the lower urinary tract identified the micturition centre functionally during electrical stimulation. Stereotactic coordinates were recorded, and the needle visualized by fluoroscopy, magnetic resonance imaging (MRI) or histologically. RESULTS: The stimulation evoked responses similar to voiding, i.e. a urethral pressure decrease followed by a bladder pressure increase; or similar to a continence manoeuvre, i.e. urethral pressure increase and no change in bladder pressure. In a few cases a continence response was evoked by stimulating a site 1 mm away from the site where a voiding response was evoked. The electrode position was detected by the fluoroscopy-based stereotactic procedure followed by subsequent MRI (one animal), and by histological analysis, verifying it to be in the dorsolateral pontine region. CONCLUSIONS: These results show that a pontine micturition centre exists in pigs similar to that described in rats, cats, dogs and humans. PMID- 15794804 TI - Gene transfer of vasoactive intestinal polypeptide into the penis improves erectile response in the diabetic rat. AB - OBJECTIVES: To determine the feasibility of transfecting penile corpora cavernosa with pcDNA3/vasoactive intestinal polypeptide (VIP) cDNA, which encodes for VIP in streptozotocin (STZ)-diabetic rats, to clarify whether transfection of VIP cDNA into the cavernosum affects the physiological response to cavernosal nerve stimulation, and whether this process would affect other organs in the diabetic rat model in vivo. MATERIALS AND METHODS: pcDNA3/VIP cDNA was injected into the corpus cavernosum of STZ-induced diabetic Sprague-Dawley rats. The intracavernosal pressure (ICP) and response to electrical stimulation of the cavernosal nerve (15 Hz, 1.5 ms, 20 V, 1 min) were measured in subsamples of rats at 1, 3, 7 and 14 days after injection; after measuring the ICP the animals were killed, and penile, hepatic, renal artery and abdominal aorta tissue samples were frozen in liquid nitrogen and stored at -80 degrees C. The gene expression of VIP in all samples, assessed as the expression of VIP mRNA, was estimated using a semiquantitative reverse-transcription polymerase chain reaction. RESULTS: The mean amplitude of ICP and expression of VIP mRNA in the cavernosa of the VIP treated rats was greater at 1, 3, 7 and 14 days after injection (P < 0.05) than in the control animals. There were no changes in the expression of VIP mRNA in hepatic, renal and abdominal aorta samples after injection (P > 0.05). CONCLUSIONS: VIP cDNA is easily incorporated into corpus cavernosum, and the expression is sustained for > or = 2 weeks in the penis in vivo. The transfer of VIP is capable of altering the physiologically relevant erectile response, as measured by an increase in the ICP after stimulating the cavernosal nerve. The intracorporal micro-injection of pcDNA3/VIP cDNA had little effect on the expression of VIP mRNA in other important organs. PMID- 15794805 TI - Assessment of microheterogeneity of blood flow in the rat urinary bladder by high resolution digital radiography. AB - OBJECTIVES: To assess high-resolution digital radiography for measuring blood flow and thus examine the microheterogeneity of bladder microcirculation in a rat model. MATERIALS AND METHODS: Microheterogeneity of blood flow in both mucosa and detrusor muscle of eight anaesthetized rats was investigated using an imaging technique with very high spatial resolution (0.1 x 0.1 mm(2)) using digital radiography combined with the deposition of (3)H-labelled desmethylimipramine. The spatial pattern of blood flow was quantified by the coefficient of variation of the regional flow (CV = sd/mean). RESULTS Muscle blood flow was less than mucous blood flow (muscle : mucosa, 2.9 : 5) in the empty bladder. In the muscle layer the blood flow distribution was more heterogeneous than that in the mucosa, with a mean (sd) CV in muscle and mucosa of 0.33 (0.033) and 0.16 (0.019), respectively (P < 0.001) at the capillary level. CONCLUSION: There was a heterogeneous distribution of blood flow in the microcirculation to capillary vessels in the muscular layer, possibly reflecting a difference in dynamic blood flow of regional perfusion of the emptied bladder. PMID- 15794806 TI - Back to the future for urological drug development? PMID- 15794807 TI - Modified tubularized transverse preputial island flap repair for severe proximal hypospadias. PMID- 15794808 TI - Tubeless and stentless percutaneous nephrolithotomy. PMID- 15794809 TI - Prostate size influences the outcome after presenting with acute urinary retention. PMID- 15794810 TI - The molecular staging of prostate cancer. PMID- 15794811 TI - Robotically assisted surgery. PMID- 15794812 TI - Comparative study of dartos fascia and tunica vaginalis pedicle wrap for the tubularized incised plate in primary hypospadias repair. PMID- 15794813 TI - Sacral ratio and fecal continence in children with anorectal malformation. PMID- 15794814 TI - Prostate cancers in the transition zone: part 2; clinical aspects. PMID- 15794815 TI - Radical retropubic prostatectomy: apical preparation and curtain dissection of the neurovascular bundle. PMID- 15794817 TI - A2 gene of Old World cutaneous Leishmania is a single highly conserved functional gene. AB - BACKGROUND: Leishmaniases are among the most proteiform parasitic infections in humans ranging from unapparent to cutaneous, mucocutaneous or visceral diseases. The various clinical issues depend on complex and still poorly understood mechanisms where both host and parasite factors are interacting. Among the candidate factors of parasite virulence are the A2 genes, a family of multiple genes that are developmentally expressed in species of the Leishmania donovani group responsible for visceral diseases (VL). By contrast, in L. major determining cutaneous infections (CL) we showed that A2 genes are present in a truncated form only. Furthermore, the A2 genomic sequences of L. major were considered subsequently to represent non-expressed pseudogenes 1. Consequently, it was suggested that the structural and functional properties of A2 genes could play a role in the differential tropism of CL and VL leishmanias. On this basis, it was of importance to determine whether the observed structural/functional particularities of the L. major A2 genes were shared by other CL Leishmania, therefore representing a proper characteristic of CL A2 genes as opposed to those of VL isolates. METHODS: In the present study we amplified by PCR and sequenced the A2 genes from genomic DNA and from clonal libraries of the four Old World CL species comparatively to a clonal population of L. infantum VL parasites. Using RT-PCR we also amplified and sequenced A2 mRNA transcripts from L. major. RESULTS: A unique A2 sequence was identified in Old World cutaneous Leishmania by sequencing. The shared sequence was highly conserved among the various CL strains and species analysed, showing a single polymorphism C/G at position 58. The CL A2 gene was found to be functionally transcribed at both parasite stages. CONCLUSION: The present study shows that cutaneous strains of leishmania share a conserved functional A2 gene. As opposed to the multiple A2 genes described in VL isolates, the CL A2 gene is unique, lacking most of the nucleotide repeats that constitute the variable region at the 5'end of the VL A2 sequences. As the variable region of the VL A2 gene has been shown to correspond to a portion of the protein which is highly immunogenic, the present results support the hypothesis of a possible role of the A2 gene in the differential tropism of CL and VL leishmania parasites. PMID- 15794818 TI - An ethical framework for cardiac report cards: a qualitative study. AB - BACKGROUND: The recent proliferation of health care report cards, especially in cardiac care, has occurred in the absence of an ethical framework to guide in their development and implementation. An ethical framework is a consistent and comprehensive theoretical foundation in ethics, and is formed by integrating ethical theories, relevant literature, and other critical information (such as the views of stakeholders). An ethical framework in the context of cardiac care provides guidance for developing cardiac report cards (CRCs) that are relevant and legitimate to all stakeholders. The purpose of this study is to develop an ethical framework for CRCs. METHODS: Delphi technique - 13 panelists: 2 administrators, 2 cardiac nurses, 5 cardiac patients, 2 cardiologists, 1 member of the media, and 1 outcomes researcher. Panelists' views regarding the ethics of CRCs were analyzed and organized into themes. RESULTS: We have organized panelists' views into ten principles that emerged from the data: 1) improving quality of care, 2) informed understanding, 3) public accountability, 4) transparency, 5) equity, 6) access to information 7) quality of information, 8) multi-stakeholder collaboration, 9) legitimacy, and 10) evaluation and continuous quality improvement. CONCLUSION: We have developed a framework to guide the development and dissemination of CRCs. This ethical framework can provide necessary guidance for those generating CRCs and may help them avoid a number of difficult issues associated with existing ones. PMID- 15794819 TI - Ocean climate and seal condition. AB - BACKGROUND: The condition of many marine mammals varies with fluctuations in productivity and food supply in the ocean basin where they forage. Prey is impacted by physical environmental variables such as cyclic warming trends. The weaning weight of northern elephant seal pups, Mirounga angustirostris, being closely linked to maternal condition, indirectly reflects prey availability and foraging success of pregnant females in deep waters of the northeastern Pacific. The aim of this study was to examine the effect of ocean climate on foraging success in this deep-diving marine mammal over the course of three decades, using cohort weaning weight as the principal metric of successful resource accrual. RESULTS: The mean annual weaning weight of pups declined from 1975 to the late 1990s, a period characterized by a large-scale, basin-wide warm decadal regime that included multiple strong or long-duration El Ninos; and increased with a return to a cool decadal regime from about 1999 to 2004. Increased foraging effort and decreased mass gain of adult females, indicative of reduced foraging success and nutritional stress, were associated with high ocean temperatures. CONCLUSION: Despite ranging widely and foraging deeply in cold waters beyond coastal thermoclines in the northeastern Pacific, elephant seals are impacted significantly by ocean thermal dynamics. Ocean warming redistributes prey decreasing foraging success of females, which in turn leads to lower weaning mass of pups. Annual fluctuations in weaning mass, in turn, reflect the foraging success of females during the year prior to giving birth and signals changes in ocean temperature cycles. PMID- 15794820 TI - Microsurgical anatomy of the dural arteries. AB - OBJECTIVE: The objective was to examine the microsurgical anatomy basic to the microsurgical and endovascular management of lesions involving the dural arteries. METHODS: Adult cadaveric heads and skulls were examined using the magnification provided by the surgical microscope to define the origin, course, and distribution of the individual dural arteries. RESULTS: The pattern of arterial supply of the dura covering the cranial base is more complex than over the cerebral convexity. The internal carotid system supplies the midline dura of the anterior and middle fossae and the anterior limit of the posterior fossa; the external carotid system supplies the lateral segment of the three cranial fossae; and the vertebrobasilar system supplies the midline structures of the posterior fossa and the area of the foramen magnum. Dural territories often have overlapping supply from several sources. Areas supplied from several overlapping sources are the parasellar dura, tentorium, and falx. The tentorium and falx also receive a contribution from the cerebral arteries, making these structures an anastomotic pathway between the dural and parenchymal arteries. A reciprocal relationship, in which the territories of one artery expand if the adjacent arteries are small, is common. CONCLUSION: The carotid and vertebrobasilar arterial systems give rise to multiple branches that supply the dura in a complex and overlapping pattern. A knowledge of the microsurgical anatomy of these dural arteries and their assessment on pretreatment evaluations plays a major role in safe and accurate treatment of multiple lesions. PMID- 15794821 TI - The triangle of the vertebral artery. AB - OBJECTIVE: Neurosurgical procedures such as proximal brachial plexus repair, scalenotomy, and direct isolation of the proximal vertebral artery require a good working knowledge of the triangle of the vertebral artery. This deep triangle of the neck is bound by the subclavian artery and the anterior scalene and longus cervicis muscles. In addition to the vertebral artery, many important structures are found in this area, such as the ganglionated sympathetic chain and certain cervical spinal nerves. METHODS: Twenty formalin-fixed cadavers were used for this study. Dissection of this triangle was performed, and measurements were made not only of parts of its borders, but also distances from these borders to neurologically important structures within its confines, such as the C8 spinal nerve. RESULTS: In all specimens, the middle scalene muscle was noted to form part of the posterior wall of the triangle. The mean height of the triangle was found to be 3.2 cm, and the mean width of its base was 1.3 cm. We observed that the C8 spinal nerve had a mean distance of 1.2 cm inferior to the apex of the triangle and that the C7 spinal nerve was found inside the triangle in 5% of sides. If the phrenic nerve entered the triangle, it was never found more than 6 mm medial to the anterior scalene muscle. The vertebral artery always traveled intimately along the lateral border of the longus cervicis muscle, and its lateral edge ranged 5 to 8 mm medial to the medial edge of the anterior scalene muscle. CONCLUSION: The C7 spinal nerve was observed in the triangle of the vertebral artery. In addition, the posterior border of the triangle of the vertebral artery was clearly defined in this study, and the middle scalene muscle could be used as a landmark. These data, coupled with our quantitation of parts and structures within the triangle, may assist neurosurgeons who operate on this area of the neck. PMID- 15794822 TI - Anatomic landmarks for nerves of the neck: a vade mecum for neurosurgeons. AB - Anatomic landmarks are often quite useful in localizing structures within the human body, especially in the surgical realm. We have reviewed the medical literature and multiple texts of surgery and anatomy and culled together many anatomic landmarks regarding important nerves of the neck. This region is complicated by formidable anatomy, and a single collection of the relationships of the nerves in this region is lacking in the literature. It is our hope that this collection will serve to assist the surgeon who operates in this region of the body. PMID- 15794823 TI - Surgical clipping of complex basilar apex aneurysms: a strategy for successful outcome using the pretemporal transzygomatic transcavernous approach. AB - OBJECTIVE: Complex basilar apex aneurysms (large size, wide and complex neck, tortuous parent vessels) continue to pose a challenge in treatment. Endovascular treatment has a high risk of recanalization, and surgical treatment is limited by the space and time necessary to achieve safe clipping. To overcome these obstacles, a modification of previously reported approaches was developed. The pretemporal transzygomatic transcavernous approach and a clipping strategy were used in the treatment of 21 high-complexity basilar apex aneurysms. METHODS: By use of the pretemporal route, the zygomatic notch was widened, the anterior clinoid was removed, the cavernous sinus was partially exposed, and the oculomotor nerve was mobilized. The depth of the field was widened by further cavernous exposure and the removal of the posterior clinoid. Temporary clips were applied to the basilar trunk perforator-free zone to preserve visualization of the aneurysm neck and perforators and to maintain collateral flow to the brainstem. RESULTS: Twenty-one high-complexity basilar apex aneurysms, 11 of which caused subarachnoid hemorrhage, were treated. Twenty (95%) were successfully clipped (Glasgow Outcome Scale scores, 4 or 5 in 90.5% at discharge; Rankin Disability Score, 1 in 90.5% at 1-yr follow-up). Complications were transient oculomotor palsy in all patients, small thalamic infarct in one patient, and cerebrospinal fluid leak in another. There was no surgical mortality. Delayed follow-up angiography in 19 of the 21 patients showed no residual aneurysm. CONCLUSION: We report the largest series of a unique, challenging group of complex basilar apex aneurysms treated with the pretemporal transzygomatic transcavernous approach, which provided improved safety of clipping by 1) increased visualization of the basilar apex and perforator arteries, 2) improved maneuverability of clip application, 3) a safer perforator free location, and 4) preservation of brainstem collateral flow. PMID- 15794824 TI - Role of uncal resection in optimizing transsylvian access to the basilar apex: cadaveric investigation and preliminary clinical experience in eight patients. AB - OBJECTIVE: The pretemporal approach has gained popularity for the treatment of basilar apex aneurysms. However, it requires the sacrifice of anterior temporal bridging veins to allow posterior temporal lobe retraction and, for patients with dominant pretemporal venous drainage, has the attendant risk of venous hypertension, hemorrhagic venous infarction, or seizures postoperatively. Alternatively, we have found that splitting the sylvian fissure, resecting the uncus, and applying posterolateral retraction to the medial temporal lobe provides a similar exposure to the basilar apex while preserving the anterior temporal bridging veins. To evaluate the transsylvian, trans-uncal approach to the basilar apex, we report our initial clinical results using this exposure in eight consecutive patients. A morphometric cadaveric analysis comparing this approach with the pretemporal approach was also performed. METHODS: For the clinical study, all hospital charts and imaging studies were retrospectively reviewed for patients undergoing the transsylvian, trans-uncal approach for the treatment of an upper basilar trunk aneurysm between July 2000 and July 2002. In the anatomic study, six formalin-fixed cadaver specimens were used. Two sequential exposures of the basilar apex were performed on each specimen side. First, the pretemporal exposure was performed with anteroposterior temporal lobe retraction. Next, after the temporal lobe had been allowed to return to normal anatomic position, the retractor was repositioned on the medial aspect of the temporal lobe superficial to the uncus, and a 10 x 10 x 15-mm volume of uncus was removed. Morphometric measurements were performed for each exposure. RESULTS: Four basilar bifurcation and four superior cerebellar segment aneurysms in eight consecutive patients were successfully clip-ligated by use of the transsylvian, trans-uncal approach. All patients had temporal bridging veins that were preserved, as documented by angiography and operative reports. No patient developed a venous infarction or new postoperative seizures, with a mean follow up of 9.75 months (range, 0.5-28 mo). The cadaveric analysis revealed that in addition to providing a similar exposure of the upper basilar complex, the transsylvian, trans-uncal approach provided additional exposure of the ipsilateral posterior cerebral and superior cerebellar arteries compared with the pretemporal approach. CONCLUSION: When approaching the basilar bifurcation, the transsylvian, trans-uncal approach provides superior exposure of the ipsilateral superior cerebellar and posterior cerebral arteries compared with the pretemporal approach, while preserving the anterior temporal bridging veins. This approach is most valuable in patients with dominant temporal venous drainage or when additional exposure of the ipsilateral posterior cerebral or superior cerebellar arteries is required. PMID- 15794826 TI - Proton magnetic resonance spectroscopic imaging integrated into image-guided surgery: correlation to standard magnetic resonance imaging and tumor cell density. AB - OBJECTIVE: In this study, we attempted to improve the delineation of the infiltration zone in gliomas using proton magnetic resonance spectroscopic imaging (1H MRSI). In conventional magnetic resonance imaging (MRI), the boundaries of gliomas sometimes are underestimated. 1H MRSI is a noninvasive tool that can be used to investigate the spatial distribution of metabolic changes in brain lesions. The purpose was to correlate tumor cell density from histopathological specimens with metabolic levels and the coregistered metabolic maps. METHODS: We developed a method to integrate spectroscopic data depicted as metabolic maps of biochemically pathological tissue into frameless stereotaxy. In seven patients harboring gliomas, we performed 1H MRSI with high spatial resolution and evaluated the spectral data. An algorithm was developed for user independent calculation of pathological voxels and for visualization as metabolic maps. These maps were integrated into a three-dimensional MRI data set used for frameless stereotaxy. Stereotactic biopsies were taken from three different areas in and around the tumor involving the maximum pathological change, the border zone, and an area from outside the spectroscopically suspicious area. These specimens were correlated to the exact voxel positions in the stereotactic image space and evaluated histopathologically. RESULTS: In all cases, the implementation of the metabolic maps into frameless stereotaxy was successful, and stereotactic biopsies were acquired by use of the spectral data. A relation could be demonstrated between the metabolic changes and tumor cell density ranging from 60 to 100% in the maximum pathological area to 5 to 15% in the border zone. Interestingly, the tumor areas defined by the metabolic maps and histopathologically confirmed by biopsy exceeded the T2-weighted signal change in all cases, ranging from 6 to 32% in the examined volume. CONCLUSION: Our preliminary data suggest that 1H MRSI may be useful in combination with frameless stereotaxy to define more exactly the tumor infiltration zone in glioma surgery compared with conventional anatomic MRI alone. PMID- 15794825 TI - Operation of arteriovenous malformations assisted by stereoscopic navigation controlled display of preoperative magnetic resonance angiography and intraoperative ultrasound angiography. AB - OBJECTIVE: To study the application of navigated stereoscopic display of preoperative three-dimensional (3-D) magnetic resonance angiography and intraoperative 3-D ultrasound angiography in a clinical setting. METHODS: Preoperative magnetic resonance angiography and intraoperative ultrasound angiography are presented as stereoscopic images on the monitor during the operation by a simple red/blue technique. Two projections are generated, one for each eye, according to a simple ray casting method. Because of integration with a navigation system, it is possible to identify vessels with a pointer. The system has been applied during operations on nine patients with arteriovenous malformations (AVMs). Seven of the patients had AVMs in an eloquent area. RESULTS: The technology makes it easier to understand the vascular architecture during the operation, and it offers a possibility to identify and clip AVM feeders both on the surface and deep in the tissue at the beginning of the operation. All 28 feeders identified on the preoperative angiograms were identified by intraoperative navigated stereoscopy. Twenty-five were clipped at the beginning of the operation. The other three were clipped at a later phase of the operation. 3-D ultrasound angiography was useful to map the size of the nidus, to detect the degree of brain shift, and to identify residual AVM. CONCLUSION: Stereoscopic visualization enhances the surgeon's perception of the vascular architecture, and integrated with navigation technology, this offers a reliable system for identification and clipping of AVM feeders in the initial phase of the operation. PMID- 15794827 TI - The initial use of free-running electromyography to detect early motor tract injury during resection of intramedullary spinal cord lesions. AB - OBJECTIVE: The resection of intramedullary spinal cord lesions (ISCLs) can be complicated by neurological deficits. Neuromonitoring has been used to reduce intraoperative risk. We have used somatosensory evoked potentials (SEPs) and muscle-derived transcranial electrical motor evoked potentials (myogenic TCE MEPs) to monitor ISCL removal. We report our retrospective experience with the addition of free-running electromyography (EMG). METHODS: Thirteen patients underwent 14 monitored ISCL excisions. Anesthesia was maintained with minimal inhalant to reduce motoneuron suppression and enhance the myogenic TCE-MEPs. Free running EMG was examined in the four limbs for evidence of abnormal bursts, prolonged tonic discharge, or sudden electrical silence. Warning of an electromyographic abnormality or myogenic TCE-MEP loss prompted interventions, including blood pressure elevation, a pause in surgery, a wake-up test, or termination of surgery. Pre- and postoperative neurological examinations determined the incidence of new deficits. RESULTS: The combined use of free running EMG and myogenic TCE-MEPs detected all eight patients with a new motor deficit after surgery; there was one false-positive report. In three of the eight true-positive cases, an electromyographic abnormality immediately anticipated loss of the myogenic TCE-MEPs. Two patients with abnormal EMGs but unchanged myogenic TCE-MEPs experienced mild postoperative worsening of motor deficits; myogenic TCE-MEPs alone would have generated false-negative reports in these cases. CONCLUSION: During resection of ISCLs, free-running EMG can supplement motor tract monitoring by TCE-MEPs. Segmental and suprasegmental elicitation of neurotonic discharges can be observed in four-limb EMG. Abnormal electromyographic bursts, tonic discharge, or abrupt electromyographic silence may anticipate myogenic TCE-MEP loss and predict a postoperative motor deficit. PMID- 15794828 TI - The role of intraoperative electromyographic monitoring in lumbosacral lipomas. AB - OBJECTIVE: To demonstrate the role of intraoperative multichannel electromyographic (EMG) monitoring to reduce postoperative deterioration and achieve full untethering of complex occult dysraphisms. METHODS: A retrospective analysis was performed on 66 patients who underwent operation for lumbosacral lipomas. Twenty recent cases were submitted to EMG monitoring and stimulation. RESULTS: All patients presented symptoms at the time of surgery, and 74% exhibited progressive deterioration during the lengthy preoperative period. Postoperative surgery-related deterioration was observed in 6% of patients. This number was reduced to zero with the introduction of intraoperative EMG monitoring. CONCLUSION: Intraoperative multichannel EMG monitoring can be carried out and requires only minimal changes to anesthetic procedures. With this method, it is possible to better identify the neural structures of complex malformations, reducing the risks of surgical damage and incomplete detethering. PMID- 15794829 TI - Lateral approach to anterolateral tumors at the foramen magnum: factors determining surgical procedure. AB - OBJECTIVE: We discuss and evaluate surgical strategies and results in 42 patients with a variety of tumors involving the anterior and anterolateral foramen magnum and present factors affecting the degree of resection and patient outcomes. We describe our surgical techniques for resection of these tumors via the lateral approach, including consideration for occipital condylar resection and vertebral artery management. METHODS: A retrospective analysis was performed of 42 surgically treated patients with tumors involving the anterior and anterolateral foramen magnum. Patients received treatment between 1991 and 2002; patients' files, operative notes, and pre- and postoperative imaging studies were used for the analysis. RESULTS: The female-to-male ratio was 28:14. Mean patient age was 47 years. Pathological entities comprised 18 meningiomas, 12 chordomas, 3 glomus tumors, 3 schwannomas, and 6 miscellaneous tumors. We mobilized the vertebral artery at the dural entry point in all patients with meningiomas. The vertebral artery was mobilized at the C1 transverse foramen for the majority of extradural tumors. Partial condyle resection was performed in eight meningiomas and five extradural tumors. Complete condyle resection was required in 12 cases, including 9 chordomas, 2 carcinomas, and 1 bone-invading pituitary adenoma. Thirteen patients required occipitocervical fusion after tumor resection. CONCLUSION: In anterior or anterolaterally located foramen magnum tumors, we think the extreme lateral or far lateral approach affords significant advantages. Vertebral artery mobilization and occipital condyle resection may be needed depending on the extent and location of the foramen magnum tumor and its specific pathological characteristics. Tumor invading the occipital condyle or significant condylar resection may cause occipitocervical instability and require fusion. PMID- 15794830 TI - Reconstruction of the cranial base in surgery for jugular foramen tumors. AB - OBJECTIVE: The surgical removal of a jugular foramen (JF) tumor presents the neurosurgeon with a complex management problem that requires an understanding of the natural history, diagnosis, surgical approaches, and postoperative complications. Cerebrospinal fluid (CSF) leakage is one of the most common complications of this surgery. Different surgical approaches and management concepts to avoid this complication have been described, mainly in the ear, nose, and throat literature. The purpose of this study was to review the results of CSF leakage prevention in a series of 66 patients with JF tumors operated on by a multidisciplinary cranial base team using a new technique for cranial base reconstruction. METHODS: We retrospectively studied 66 patients who had JF tumors with intracranial extension and who underwent surgical treatment in our institutions from January 1987 to December 2001. Paragangliomas were the most frequent lesions, followed by schwannomas and meningiomas. All patients were operated on using the same multidisciplinary surgical approach (neurosurgeons and ear, nose, and throat surgeons). A surgical strategy for reconstruction of the cranial base using vascularized flaps was carried out. The closure of the surgical wound was performed in three layers. A specially developed myofascial flap (temporalis fascia, cervical fascia, and sternocleidomastoid muscle) associated to the inferior rotation of the posterior portion of the temporalis muscle was used to reconstruct the cranial base with vascularized flaps. RESULTS: In this series of 66 patients, postoperative CSF leakage developed in three cases. These patients presented with very large or recurrent tumors, and the postoperative CSF fistulae were surgically closed. The cosmetic result obtained with this reconstruction was classified as excellent or good in all patients. CONCLUSION: Our results compare favorably with those reported in the literature. The surgical strategy used for cranial base reconstruction presented in this article has several advantages over the current surgical techniques used in cases of JF tumors. PMID- 15794831 TI - Combination of functional magnetic resonance imaging-guided neuronavigation and intraoperative cortical brain mapping improves targeting of motor cortex stimulation in neuropathic pain. AB - OBJECTIVE: To evaluate, regardless of the clinical results, the contribution of combining functional magnetic resonance imaging (fMRI) with intraoperative cortical brain mapping (iCM) as functional targeting methods for epidural chronic motor cortex stimulation (MCS) in refractory neuropathic pain. METHODS: Eighteen neuropathic pain patients (central stroke in six; trigeminal neuropathy in six; syrinx or amputation in six) who underwent operations for epidural MCS were studied with preoperative fMRI and iCM. fMRI investigated motor tasks of hands (as well as foot and tongue, when painful). fMRI data were analyzed with Statistical Parametric Mapping99 software (University College London, London, England; initial analysis threshold corresponding to P < 0.001), registered in a neuronavigation system, and correlated during surgery with iCM. The primary aim of this study was to improve the topographical precision of MCS. Matching of fMRI and iCM specifically was examined. RESULTS: Correspondence between the contour of the fMRI activation area and iCM in precentral gyrus (mean distance, 3.8 mm) was found in 17 (94%) of 18 patients. Eleven of them showed correspondence for more restrictive values of the analysis threshold (P < 0.0001); in six patients, the quality of the iCM was reduced by somatosensory wave attenuation and general anesthesia. In this group of six patients, a combination of both techniques was used for the final targeting. Correspondence was not found in one patient as the result of image distortion and residual motion artifact. At follow-up (4-60 mo), MCS induced significant pain relief in a total of 11 patients (61%). CONCLUSION: This study confirms the functional accuracy of fMRI guidance in neuropathic pain and illustrates the usefulness of combining fMRI guidance with iCM to improve the functional targeting in MCS. Because appropriate targeting is crucial to obtaining pain relief, this combination may increase the analgesic efficacy of MCS. PMID- 15794832 TI - Comparison of three methods of targeting the subthalamic nucleus for chronic stimulation in Parkinson's disease. AB - OBJECTIVE: The success of subthalamic nucleus (STN) surgery for Parkinson's disease depends on accuracy in target determination. The objective of this study was to determine which of the following techniques was most accurate and precise in identifying the location for stimulation in STN deep brain stimulation surgery that is most clinically effective: direct targeting, indirect targeting using the positions of the anterior and posterior commissures, or a technique using the red nucleus (RN) as an internal fiducial marker. METHODS: We reviewed 14 patients with Parkinson's disease treated with bilateral STN deep brain stimulation (28 STN targets). Electrode implantation was based on direct and indirect targeting using two-dimensional magnetic resonance imaging with refinement using microelectrode recording. Optimal settings, including the contacts used, were determined during the clinical follow-up. The position of the best contact was defined with postoperative magnetic resonance imaging. This location was compared with the modified direct, indirect, and RN-based targets. The mean distances between the targets and the final position of the optimal contact were calculated. The accuracy and variance of each target were analyzed. RESULTS: The mean position of the best contact was x = 12.12 (standard deviation [SD], 1.45 mm), y = -2.41 (SD, 1.63 mm), and z = -2.39 (SD, 1.49 mm) relative to the midcommissural point. The mean distance between the optimal contact position and the planned target was 3.19 mm (SD, 1.19 mm) using the RN-based method, 3.42 mm (SD, 1.34 mm) using indirect targeting, and 4.66 mm (SD, 1.33 mm) using a modified direct target. The mean distance between the optimal contact and the RN based target was significantly smaller than the mean distance between the optimal contact and the direct target (post hoc with Tamhane's correction, P < 0.001) but not between the optimal contact and the indirect target. The RN-based target had the smallest variance (F test, P < 0.001), indicating greater precision. CONCLUSION: The use of the RN as an internal fiducial marker for targeting the optimal region of STN stimulation was reliable and closely approximates the position of the electrode contact that provides the optimal clinical results. PMID- 15794833 TI - Coverage of painful peripheral nerve neuromas with vascularized soft tissue: method and results. AB - OBJECTIVE: Our goals are to describe a method of treating painful peripheral nerve neuromas by means of vascularized tissue coverage, report the results in seven patients, and discuss the indications for this treatment modality. An analysis of pain, functionality of the affected body part, professional activities of the patients, and medications before and after surgery is presented. METHODS: Seven male patients (mean age, 45.1 yr) with posttraumatic nerve injuries, who had developed painful stump neuromas or neuromas-in continuity, and who had unsuccessfully undergone several treatment procedures, were selected for the surgery described here. The operation included resection of the stump neuroma (four patients) or neurolysis of the neuroma-in-continuity (three patients) and coverage of the nerve with a vascularized fascial, fasciocutaneous, or perforator flap (three pedicled regional flaps and four free flaps). A modified quadruple visual analog scale was used to quantify pain before and after surgical treatment. The mean follow-up was 16.6 months. RESULTS: The mean values of the quadruple visual analog scale (pain now/typically/at its best/at its worst) before surgery were 6.5/6.5/4.7/7.9. These values changed to 0.3/0.4/0/0.9 at a mean follow-up of 16.6 months after surgery. Five patients returned to their original profession, one receives a pension, and one began a less demanding job after undergoing surgery. Six of the seven patients received opioids before surgery (one had a spinal cord stimulator). After surgery, all patients stopped taking regular pain killers and the spinal cord stimulator was deactivated in one; two patients still take nonsteroidal anti-inflammatory drugs occasionally, but not on a regular basis. CONCLUSION: Vascularized soft tissue coverage of painful peripheral nerve neuromas seems to be an effective and attractive, but also complex, method of treatment. This option may be considered and reserved for patients who have already undergone several pain treatment modalities without success. PMID- 15794834 TI - Endoscopic transnasal approach to the cavernous sinus versus transcranial route: anatomic study. AB - OBJECTIVE: The aim of the present study was to compare the anatomy of the cavernous sinus via an endoscopic transnasal route with the anatomy of the same region explored by the transcranial route. The purpose was to identify and correlate the corresponding anatomic landmarks both through the endoscopic transnasal transsphenoidal and the microscopic transcranial views. METHODS: Five fresh injected heads (10 specimens) were dissected by the endoscopic transnasal and microsurgical transcranial approaches. A comparison of different microsurgical corridors of the cavernous sinus with the corresponding endoscopic transnasal ones was performed. RESULTS: Through the endoscopic transnasal approach, it is possible to explore only some of the parasellar and middle cranial fossa subregions. Because of the complex multilevel architecture of the cavernous sinus, there is not always a correspondence between the surgical corridors bounded through the transcranial route and those exposed through the endoscopic transnasal approach. Nevertheless, some surgical corridors specific to the endoscopic transnasal route are evident: a C-shaped corridor is identifiable medial to the "intracavernous" internal carotid artery, whereas a wider triangular area is delineable lateral to the internal carotid artery; inside the latter, three more surgical corridors (a superior triangular space, a superior quadrangular space, and an inferior quadrangular space) can be described. CONCLUSION: Different surgical corridors can be defined during the endoscopic transnasal approach to the anteroinferior portion of the cavernous sinus, as already established for the transcranial route as well. Knowledge of these could be useful in decreasing morbidity and mortality during surgery in this region, these approaches being reserved to experienced transsphenoidal surgeons only. PMID- 15794835 TI - Microsurgical anatomy of the transcallosal anterior interforniceal approach to the third ventricle. AB - OBJECTIVE: We explored relevant regional microanatomy as it relates to the challenging anterior interforniceal (AIF) approach for removing hypothalamic hamartomas. METHODS: Five silicone-injected cadaveric heads were dissected by use of frameless stereotactic navigation to reveal microanatomy and extent of exposure through the transcallosal AIF approach. Distances between trajectories to the coronal suture and the genu of the corpus callosum (CC) and between the posterior border of the anterior commissure to the lower end of the rostrum of the CC and posterior border of the foramen of Monro were measured. RESULTS: The AIF approach provided adequate access to the anterior third ventricle and related structures (i.e., hypothalamus, infundibular recess, and mamillary bodies) through the corridor bounded by the anterior commissure anteriorly and the choroid plexus at the foramen of Monro posteriorly. The mean distances from the posterior trajectory to the coronal suture and the genu of the CC were 44.8 mm (range, 43.8-46.2 mm) and 14.88 mm (14.1-15.7 mm), respectively. The mean distance from the anterior trajectory posterior to the coronal suture was 4.66 mm (0-8.9 mm), and 32.6 mm (30.5-33.9 mm) to the genu of the CC. The mean length of callosotomy was 17.52 mm (16.2-19.1 mm). The mean distance between the posterior border of the anterior commissure and the lower end of the rostrum of the CC was 5.22 mm (4.6-5.6 mm), and 10.52 mm (9.7-11.5 mm) to the posterior border of the foramen of Monro. CONCLUSION: The technically safe AIF approach permitted limited interforniceal splitting, no major deep vein manipulation, and adequate visualization of the hypothalamus, infundibular recess, and mamillary bodies. PMID- 15794836 TI - Quantitative anatomic study of three surgical approaches to the anterior communicating artery complex. AB - OBJECTIVE: To compare the angles of approach and area of exposure to the anterior communicating artery (AComA) complex associated with pterional (PT), orbitopterional (OPT), and orbitozygomatic (OZ) craniotomies before and after gyrus rectus resection. METHODS: PT, OPT, and OZ craniotomies were performed on both sides of four heads, and the angles of approach and area of exposure to the AComA complex were measured before and after resection of the gyrus rectus. RESULTS: The vertical angle of approach increased significantly among the PT and OPT (P = 0.001), PT and OZ (P = 0.001), and OPT and OZ approaches (P = 0.005). The horizontal angle of approach was significantly larger between the PT to OPT (P = 0.001) and PT to OZ (P = 0.001) approaches but not between the OPT and OZ approaches (P = 0.757). After gyrus rectus resection, the vertical and horizontal angles of approach increased significantly for the PT approach but not for the OPT and OZ approaches. The area of exposure to the AComA complex increased progressively from the PT to OPT to OZ approach but did not reach statistical significance (P = 0.124). Resection of the gyrus rectus resulted in significant relative gains in the area of exposure for the PT (P = 0.01) and OPT (P = 0.04) approaches but not for the OZ approach (P = 0.88). CONCLUSION: The vertical and horizontal angles of approach to the AComA complex are significantly larger for the OPT and OZ approaches compared with the PT approach. Use of the OZ approach may decrease the need for frontal lobe retraction and resection of the gyrus rectus. PMID- 15794838 TI - The microsurgical nuances of resecting tuberculum sellae meningiomas. AB - In a recent article, our experience and knowledge of the clinical picture, microsurgical anatomy, and long-term surgical outcome of resecting tuberculum sellae meningiomas was described in detail. We now present our surgical technique in a pictorial and video format for the benefit of neurosurgeons in training, as well as for general critique. Attention is given to the details of surgery: patient positioning, surgical approaches, technique of tumor removal, and postoperative care. PMID- 15794837 TI - Anterior ethmoidal artery: microsurgical anatomy and technical considerations. AB - OBJECTIVE: Vascular lesions of the anterior cranial fossa can receive significant blood supply from the anterior ethmoidal artery. Embolization of this blood supply exposes the parent vessel, the ophthalmic artery, to possible embolic complications, which can lead to loss of vision. A study of the microsurgical anatomy can help delineate the course of the anterior ethmoidal artery and find the best points for proximal control of the blood supply to these lesions. Clinical cases are presented to illustrate how lesions with prominent anterior ethmoidal artery feeders are best approached through fronto-orbital single-flap craniotomies. METHODS: Eight cadaveric dissections to demonstrate the microsurgical anatomy of the anterior ethmoidal artery were performed to study the relevant anatomy. Two clinical cases are presented that demonstrate clinical application of this anatomy through fronto-orbital single-flap craniotomies. RESULTS: Eight arteries were studied in four cadaveric heads. The dissections show the course of the anterior ethmoidal artery from the ophthalmic artery in the orbit, through the anterior ethmoidal foramen into the ethmoid air cells, to the cribriform plate, where it turns superiorly to become the anterior falx artery. The first surgical case is of a giant tuberculum sellae meningioma that was resected with coagulation and division of the anterior ethmoidal arteries at the anterior ethmoidal foramina at the laminae papyraceae of both medial orbital walls. The second surgical case is of a large deep right frontal arteriovenous malformation that was resected with coagulation and division of the anterior ethmoidal artery at the anterior ethmoidal foramen of the lamina papyracea of the right medial orbital wall. CONCLUSION: The cadaveric dissections and our surgical experience show that the anterior ethmoidal artery has three important sites for surgical access: 1) the anterior ethmoidal foramen at the lamina papyracea of the medial orbital wall; 2) the anterior ethmoid canal at the lateral ethmoid wall; and 3) extradurally, at the cribriform plate. These three sites are best accessed through a fronto-orbital single-flap craniotomy, which can be unilateral or bilateral, depending on the pathological findings. The described orbital-cranial approach in this article is not being advocated to replace the standard pterional and frontal approaches; rather, we suggest it as an option in these complex cases that require early proximal control of the anterior ethmoidal artery feeders. PMID- 15794839 TI - Frontobasal interhemispheric trans-lamina terminalis approach for suprasellar lesions. AB - The frontobasal interhemispheric approach for suprasellar tumors currently incorporates technological advancements and refinements in patient selection, operative technique, and postoperative care. This technique is a valid choice for the removal of suprasellar lesions with extension into the third ventricle without major sequelae related to the surgical approach. The method described here reflects the combination of the frontal interhemispheric and trans-lamina terminalis approaches. PMID- 15794840 TI - Reduction of postoperative perineural adhesions by Hyaloglide gel: an experimental study in the rat sciatic nerve. AB - OBJECTIVE: To assess the effects of Hyaloglide gel (or auto-cross-linked polysaccharide [ACP] gel; Fidia Advanced Biopolymers, Abano Terme, Italy), a hyaluronan-derivative polymer, on peripheral nerve scarring and nerve regeneration. METHODS: We performed two surgical procedures in adult rats: 1) neurolysis of the sciatic nerve and separation of its tibial and peroneal branches, and 2) transection and immediate suture of the sciatic nerve. After nerve manipulation, ACP gel was applied onto the site of operation. We tested two solutions of ACP gel having different viscosities. Additional animals received Adcon-T/N (Gliatech, Inc., Cleveland, OH), an antiadhesive agent currently available for clinical use. No gel was applied on the contralateral side, which served as a control side. Four weeks later, the animals underwent reoperation. We assessed the quality of wound healing, the presence of perineural adherences, and the separability of nerves from surrounding tissues. RESULTS: Significantly fewer perineural adhesions were found in animals treated with ACP gel (high viscosity) and Adcon-T/N compared with controls. Quantitative histological analysis revealed a statistically significant reduction in the amount of scar tissue surrounding the nerves treated with ACP gel. No evidence of toxicity was found, and the gel did not interfere with nerve regeneration (counts of regenerating myelinated axons). CONCLUSION: ACP gel with high viscosity seems to be safe and effective in reducing perineural adhesions and scar formation after peripheral nerve surgery. PMID- 15794841 TI - Intuitive intraoperative ultrasound guidance using the Sonic Flashlight: a novel ultrasound display system. AB - OBJECTIVE: The Sonic Flashlight (SF) is a new handheld ultrasound (US) display device being developed at our institution. It replaces the standard monitor on a conventional ultrasound (CUS) system with a miniature monitor and half-silvered mirror to reflect real-time US images into the body. With the SF, the imaged body part appears translucent, with the US image appearing to float below the surface of the anatomy, exactly where it is being scanned. The SF merges the patient, US image, instrument, and operator's hands into the same field of view, allowing the user to operate directly on the US image using normal hand-eye coordination. In contrast, CUS procedures result in displaced hand-eye coordination when the operator looks away from the patient to view the CUS monitor. Intraoperatively, the SF may make localizing and accessing tumors, foreign bodies, hematomas, vascular malformations, and ventricles easier and more accurate, especially for those without extensive CUS training. METHODS: In this cadaver study, the SF was used to visualize the brain and guide a needle into an implanted simulated tumor. The needle was inserted both in the US plane and outside of the US plane. INSTRUMENTATION: Sonic Flashlight fifth generation research prototype. CONCLUSION: The needle was easily and intuitively visualized and guided into the lesion, both within and outside of the US plane. By having the US image appear directly beneath the brain surface, the surgeon can easily and quickly guide the needle or surgical instrument to the lesion. The operator's eyes never have to leave the surgical field, as they do with CUS technology. The impact of this device on neurosurgical procedures could be significant. The ease of use, intuitive function, and small instrument size allow the surgeon to quickly localize lesions, confirm surgical positioning, and assess postoperative results. PMID- 15794842 TI - Lateral transsulcal approach to asymptomatic trigonal meningiomas with correlative microsurgical anatomy: technical case report. AB - OBJECTIVE: We introduce the lateral transsulcal approach to asymptomatic trigonal meningiomas. METHODS: The approach was studied in two cadaver brains and three asymptomatic patients with trigonal meningiomas. The posterior part of the sylvian fissure, or superior temporal sulcus, is opened to the bottom. Through a small horizontal cortical incision, the trigone of the lateral ventricle is exposed in the shortest distance. The trigonal meningiomas are detached from the choroid plexus and removed. RESULTS: In patients with meningiomas on the nondominant side, the transsylvian approach was adopted. In patients with meningiomas on the dominant side, the transsylvian approach was adopted for patients with a wide sylvian cistern, and the approach through the superior temporal sulcus was adopted for patients with a narrow sylvian cistern. The transverse gyrus of Heschl was a good anatomic landmark in the operative field of the transsylvian approach. Patients with meningiomas on the dominant side exhibited transient amnestic aphasia and dyscalculia, but the symptoms disappeared in a few days or weeks. These patients were discharged without any neurological deficits. Although there are potential risks of damaging association fibers, optic radiation, the transverse gyrus of Heschl, and the parietal lobe, a thorough understanding of the topographical anatomy and careful dissection techniques can avoid morbidity. Wide opening of the sylvian fissure and debulking of the tumor are other important factors to reduce the retraction of the parietal and temporal lobes. CONCLUSION: The lateral transsulcal approach is applicable for small asymptomatic trigonal meningiomas with an acceptable risk of morbidity, even in the dominant hemisphere. PMID- 15794843 TI - Intraoperative motor mapping of the cerebral peduncle during resection of a midbrain cavernous malformation: technical case report. AB - OBJECTIVE AND IMPORTANCE: Brainstem cavernous malformations that seem to come to a pial or ependymal surface on preoperative magnetic resonance imaging studies may, in fact, be covered by an intact layer of neural tissue. For cavernous malformations in the cerebral peduncle, intraoperative stimulation mapping with a miniaturized probe can determine whether this overlying tissue harbors fibers in the corticospinal tract. In addition, intermittent monitoring with transcranial motor evoked potentials (TcMEPs) helps to protect this vital pathway during resection of the lesion. CLINICAL PRESENTATION: A 20-year-old woman collapsed after a cavernous malformation in the left cerebral peduncle hemorrhaged into the pons, midbrain, and thalamus. She presented with right hemiparesis and left oculomotor palsy. INTERVENTION: The cavernous malformation was completely resected through a left orbitozygomatic craniotomy and transsylvian approach. Stimulation mapping of the cerebral peduncle with a Kartush probe (Medtronic Xomed, Inc., Jacksonville, FL) identified the corticospinal tract lateral to the lesion, and a layer of tissue over the lesion harbored no motor fibers. TcMEP monitoring helped to guide the resection, with increased voltage thresholds and altered waveform morphologies indicating transient impaired motor conduction. All TcMEP changes returned to baseline by the end of the procedure, and the patient's hemiparesis improved after surgery. CONCLUSION: Stimulation mapping of the corticospinal tract and intermittent TcMEPs is a safe and simple surgical adjunct. Expanded monitoring of the motor pathway during the resection of cerebral peduncle cavernous malformations may improve the safety of these operations. PMID- 15794844 TI - Intraoperative awakening for vision examination during ophthalmic artery aneurysm clipping: technical case report. AB - OBJECTIVE AND IMPORTANCE: We present a case of a patient with an ophthalmic artery aneurysm in which the ophthalmic artery originated from the body of the aneurysm, requiring sacrifice of the ophthalmic artery to achieve complete aneurysm obliteration. We awakened the patient intraoperatively to assess optic nerve function after clipping and were able to confirm optic nerve function. Controlled intraoperative awakening proved a valuable adjunct to intraoperative angiography in determining the immediate consequences of sacrifice of the ophthalmic artery. CLINICAL PRESENTATION: The patient was a 55-year-old right handed woman with a 3-month history of episodic blurriness in her left eye; imaging demonstrated an unruptured 5-mm left ophthalmic artery aneurysm in which the ophthalmic artery originated from the body of the aneurysm. INTERVENTION: Complete obliteration of the aneurysm required clip placement across the neck of the aneurysm, incorporating not only the aneurysm but also the ophthalmic artery. Aware that sacrifice of the ophthalmic artery was likely, we awakened the patient after clipping and before dural closure to evaluate her optic nerve function. Once fully awake, the patient was able to execute simple commands and conclusively confirm light perception in both of her eyes. She was then reanesthetized, and intraoperative angiography showed successful aneurysm obliteration and parent artery patency. CONCLUSION: The ophthalmic artery can be sacrificed during aneurysm clipping without loss of vision in many cases, most likely because of adequate collateral filling from the external carotid artery. Certainty about the visual consequences of sacrifice of the ophthalmic artery, however, is difficult to obtain preoperatively or intraoperatively. Intraoperative awakening for evaluation of optic nerve function served as a useful technique to assess the acute results of interruption of ophthalmic artery flow in this case. PMID- 15794845 TI - Transaxillary carotid stenting: technical case report. AB - OBJECTIVE AND IMPORTANCE: As endovascular neurointerventions continue to evolve rapidly, angioplasty and stenting of both the extracranial and intracranial vessels have become more routine procedures. When the transfemoral approach is contraindicated or technically difficult, familiarity with alternative access techniques becomes essential. We report a successful transaxillary carotid stenting in a patient with an axillary bifemoral bypass graft. CLINICAL PRESENTATION: A 77-year-old man presented with a symptomatic high-grade stenosis (80%) of the left internal carotid artery. Because of the increased risk of general anesthesia related to his advanced age and severe comorbidities, stenting of the left internal carotid artery was considered. A left transaxillary approach was chosen because of the presence of an axillary bifemoral bypass graft. TECHNIQUE: Under ultrasound guidance, the left axillary artery was successfully punctured and cannulated. After a 0.038 Magic Torque wire (Boston Scientific/Medi Tech, Watertown, MA) was anchored with the tip of the wire in the distal left occipital artery, a 7-French (outer diameter) Vista Bright guiding sheath (Cordis, Miami, FL) was successfully positioned in the mid left common carotid artery, with an MPA catheter (Cordis) used as guiding support. Subsequently, two Precise stents (Cordis) were successfully deployed across the stenosis, yielding a satisfactory angiographic result. CONCLUSION: With proper patient selection and the use of ultrasound guidance during the initial puncture, the transaxillary approach is a safe and technically feasible alternative to the transfemoral approach when performing carotid stenting. PMID- 15794846 TI - Temporary aneurysm orifice balloon occlusion as an alternative to retrograde suction decompression for giant paraclinoid internal carotid artery aneurysms: technical note. AB - OBJECTIVE: Giant paraclinoid carotid artery aneurysms frequently require the temporary interruption of local circulation to facilitate safe clip occlusion. Owing to the brisk retrograde blood flow through the ophthalmic artery and cavernous branches, the simple trapping of the aneurysm by cervical internal carotid artery clamping and intracranial distal clipping may not adequately soften the lesion. Retrograde suction decompression aspiration of this collateral supply by a catheter introduced into the cervical internal carotid artery is a popular method to achieve aneurysm deflation. With a large collateral supply, the method is not effective enough. The advent of relatively long and maneuverable soft balloons allows temporary occlusion of the aneurysm orifice. METHODS: We applied this method in two instances of giant carotid ophthalmic aneurysms. In both instances, a 15- to 20-mm-long and 4-mm-wide occlusion balloon was inserted in the internal carotid artery at the level of the aneurysm before craniotomy. After craniotomy and dissection of the aneurysm neck, the balloon was inflated under intraoperative angiographic control. RESULTS: The aneurysm became soft enough in both cases without tapping and aspiration to allow safe clip occlusion. In the first case, the postoperative course was uneventful and visual acuity improved. A known additional infraclinoid part of the aneurysm was eliminated endovascularly 5 months later using balloon-protected injection of vinyl alcohol copolymer (Onyx; Micro Therapeutics, Inc., Irvine, CA). In the second case, a postoperative symptomatic vasospasm developed 15 hours after surgery. Hypertensive therapy resulted in the disappearance of symptoms and an otherwise uneventful course with improvement of vision. CONCLUSION: This preliminary experience suggests that this new method is a feasible alternative to retrograde suction decompression. PMID- 15794847 TI - Endonasal transsphenoidal transclival removal of prepontine epidermoid tumors: technical note. AB - OBJECTIVE: Prepontine retroclival tumors have typically been removed through a variety of anterolateral, lateral, and posterolateral cranial base approaches. Here, we describe an endonasal transclival cranial base approach for removal of prepontine epidermoid tumors. METHODS: Two men, 40 and 52 years old, each presented with a history of headaches and cranial nerve deficits. In each patient, magnetic resonance imaging showed a large T1 hypointense/T2 hyperintense mass occupying the posterior suprasellar, premesencephalic, and prepontine cisterns, with significant mass effect on the brainstem. Both patients underwent an endonasal transsphenoidal transclival cranial base tumor removal with the operating microscope and endoscopic assistance. The dural and bony defects were repaired with abdominal fat grafts, collagen sponge, titanium mesh, and cerebrospinal fluid diversion. One patient developed a postoperative cerebrospinal fluid leak and meningitis requiring two reoperations to repair, ultimately with fat and fascia lata grafts. RESULTS: At 1 year after surgery, both patients have improved compared with their preoperative neurological state, and volume analysis of preoperative and 1-year postoperative magnetic resonance imaging scans confirm a marked reduction in mass effect on the brainstem, with a 78% tumor removal in one patient and 76% removal in the other. Both patients have normal endocrine function. CONCLUSION: The endonasal approach offers a minimally invasive, anatomically direct route for removing prepontine epidermoid tumors that obviates brain retraction. The use of angled endoscopes is essential for gaining lateral, cephalad, and caudal visualization to augment the limited microscope view. Inadequate repair of clival dural defects remains the greatest potential pitfall in attempting transsphenoidal transclival tumor removal. PMID- 15794848 TI - Minimally invasive, endoscopic, internal thoracoplasty for the treatment of scoliotic rib hump deformity: technical note. AB - OBJECTIVE: Patients with idiopathic scoliosis often have a noticeable rib deformity that frequently persists after corrective surgery. Open thoracoplasty has been the traditional method of reducing rib deformity. Recently, however, video-assisted thoracoscopy (VATS) has been used to perform thoracoplasty. There have been no long-term follow-up studies on VATS thoracoplasty, nor have there been outcome scores to assess the results of thoracoplasty procedures. We present our experience using VATS thoracoplasty with long-term follow-up and propose an outcome grading system for thoracoplasty. METHODS: Between 1998 and 2000, four patients (age range, 14-53 yr) underwent VATS thoracoplasty for significant rib hump deformity (mean height, 5 cm; range, 4-6 cm) associated with idiopathic scoliosis. All patients had four rib segments resected during the VATS thoracoplasty procedure. Three of the four patients also underwent anterior thoracic release and discectomy during the procedure. RESULTS: Patients were followed for a mean of 40 months after surgery (range, 33-50 mo). There were no intraoperative or postoperative complications. Outcomes were assessed using a patient questionnaire with our new thoracoplasty grading system. All patients were pleased that they had chosen to have VATS internal thoracoplasty. Based on our new grading system, two patients had an excellent outcome and two had a good outcome. CONCLUSION: VATS provides an alternative, minimally invasive route to perform thoracoplasty. VATS incisions are much smaller and more cosmetically appealing than open thoracoplasty incisions. Long-term follow-up indicates good to excellent patient outcomes. PMID- 15794849 TI - Ventralis intermedius plus ventralis oralis anterior and posterior deep brain stimulation for posttraumatic Holmes tremor: two leads may be better than one: technical note. AB - OBJECTIVE AND IMPORTANCE: To describe the effects of ventralis oralis anterior (VOA) and posterior (VOP), as well as ventralis intermedius (VIM), deep brain stimulation (two ipsilateral thalamic leads implanted) on posttraumatic Holmes tremor. Results of both thalamic lesioning and thalamic deep brain stimulation for Holmes tremor and tremors due to posttraumatic lesions in the region of the midbrain have been disappointing. In 2001, the use of two electrodes implanted in parallel for severe essential tremor was reported. We propose the use of a similar technique for posttraumatic Holmes tremor. One rationalization for the placement of two leads was to affect both the cerebellar receiving area (VIM) and the pallidal receiving area (VOA/VOP). A second rationalization was that the placement of a second electrode may affect somatotopy, and may, therefore, be beneficial for the treatment of more difficult to control tremor subtypes. CLINICAL PRESENTATION: A 24-year-old man with intractable posttraumatic Holmes tremor presented for consideration of a surgical intervention. INTERVENTION: A high-resolution, volumetric magnetic resonance imaging scan was obtained 1 day before the procedure. Microelectrode recording was used in addition to stereotactic computed tomography, image fusion, and stereotactic targeting to map the locations of the VIM, VOP, and VOA nuclei of the thalamus. A deep brain stimulation electrode was then implanted on the border between the left VIM and VOP thalamic nuclei, and a second ipsilateral deep brain stimulation lead was placed on the VOA and VOP border, 2 mm anterior to the first. Fourteen videotaped tremor rating scales were evaluated by two blinded reviewers. CONCLUSION: The patient experienced tremor rebound with VIM-VOP monotherapy. However, when the second lead (VOA/VOP) was activated, he experienced sustained improvement in tremor and tremor disability at a 12-month follow-up examination. This case elucidates a potential new approach for the treatment of patients with posttraumatic Holmes tremor. Additional study and longer follow-up periods will be needed to further evaluate this promising therapy. PMID- 15794850 TI - Report of the Third International Workshop on Waldenstrom's macroglobulinemia. PMID- 15794851 TI - Origins of Waldenstrom's macroglobulinemia: does it arise from an unusual B-cell precursor? AB - Clonotypic B cells of Waldenstrom's macroglobulinemia (WM) are CD20+ immunoglobulin (Ig) M+ IgD+ cells that lack ongoing somatic hypermutation and class switch recombination (CSR). Only a small compartment of clonotypic B cells express activation-induced cytosine deaminase. Activation by CD40L/interleukin-4 does not stimulate WM class switching. However, we found that the mutation of switch regions essential for CSR were present in IgM monoclonal gammopathy of unknown significance (MGUS) but absent from WM B cells, suggesting the possibility that not all IgM MGUS have the potential to give rise to WM, and further strengthening the hypothesis that the target cell in transformation to WM is an unusual type of B cell. PMID- 15794853 TI - Autoantibody activity in Waldenstrom's macroglobulinemia. AB - Some monoclonal proteins from patients with Waldenstrom's macroglobulinemia (WM) or immunoglobulin (Ig) M monoclonal gammopathy of undetermined significance possess antigen-binding activity directed to autogenous or foreign antigens. These monoclonal IgM autoantibodies include cold agglutinins, mixed cryoglobulins, and antineural components. Because of the antigenantibody interaction, patients with these autoimmune syndromes often present with hemolytic anemia, mixed cryoglobulinemia, or peripheral neuropathy, respectively, at an earlier stage than patients with typical WM who do not have evident antibody activity. The presence of monoclonal macroglobulin autoreactive antibodies thus influences clinical presentation and natural history. Monoclonal IgM antibodies display polyreactivity to antigens of microbial origin in addition to autogenous antigens and may arise through T-independent as well as T-dependent pathways. Waldenstrom proteins with antibody activity appear to provide a link between autoimmunity, infection, and lymphoproliferative disease. Study of the antigens reacting with monoclonal IgMs may provide further insight into the pathogenesis of WM. PMID- 15794852 TI - Clinicopathologic features of Waldenstrom's macroglobulinemia and marginal zone lymphoma: are they distinct or the same entity? AB - Waldenstrom's macroglobulinemia (WM) is considered in the World Health Organization classification as a clinical syndrome associated with monoclonal immunoglobulin (Ig) M secretion, mainly observed in patients with lymphoplasmacytic lymphoma (LPL) and occasionally with other small B-cell lymphomas. Some authors consider it a rare distinct lymphoproliferative disorder with primary bone marrow infiltration and IgM monoclonal gammopathy. As LPL shares important morphologic and immunophenotypic overlaps with marginal zone B cell lymphomas (MZLs) in cases showing plasmacytic maturation, it remains unclear if they constitute unique or distinct entities. Both diseases are composed of lymphocytes, lymphoplasmacytoid cells, and tumoral plasma cells with a surface (s) IgM-positive sIgD+/ cytoplasmic IgMpositive CD19+ CD20+ CD27+/ CD5 CD10 CD23 phenotype, without a specific marker. Extranodal mucosa-associated lymphoid tissue (MALT) lymphoma, nodal MZL (NMZL), and splenic MZL (SMZL) are distinct entities displaying common morphologic, immunophenotypic, and genetic characteristics. MALT lymphoma is clearly distinct from LPL, although bone marrow infiltration and IgM paraprotein are not rare. Splenic MZL and NMZL are incompletely characterized, but a plasmacytoid/plasmacytic differentiation, autoimmune manifestations, and monoclonal component are frequent in both diseases. Bone marrow involvement is constant in SMZL and present in 60% of NMZLs. Molecular IgVH gene analysis has confirmed this heterogeneity, particularly within SMZL, with mutated and unmutated cases. Further studies are needed to clarify the pathogenesis of these MZLs and their relationship with LPL. PMID- 15794854 TI - Cytogenetics of familial Waldenstrom's macroglobulinemia: in pursuit of an understanding of genetic predisposition. AB - Despite recent identification of a recurrent chromosome 6q21 deletion in sporadic Waldenstrom's macroglobulinemia (WM), elucidation of the molecular pathogenesis of WM remains challenging. In contrast to the growing body of cytogenetic studies in sporadic WM, there have been virtually no informative studies of familial WM. The authors therefore undertook conventional and molecular cytogenetic evaluation of 18 patients with familial WM and 3 patients with immunoglobulin (Ig) M monoclonal gammopathy (IgM-MG) from 15 families to determine the nature and extent of chromosomal abnormalities associated with familial WM. The frequency and distribution of chromosomal changes in familial WM resembled those in sporadic WM, including lack of IgH rearrangements and t(9;14); however, we detected del6q21 in only 1 patient. Occasional findings appeared to be novel; however, none were recurrent, and their significance remains unclear. Only one abnormality found in bone marrow specimens was detected in parallel peripheral blood lymphocyte studies, suggesting that most abnormalities represented somatic changes. Although they must be viewed in light of the hypoproliferative nature of WM, our results suggest that further progress in delineating the genetic determinants of WM susceptibility might be gained from alternative approaches such as candidate gene or linkage analysis. PMID- 15794855 TI - Differential diagnosis of Waldenstrom's macroglobulinemia and other B-cell disorders. AB - Waldenstrom's macroglobulinemia (WM) is characterized by lymphoplasmacytic infiltration of bone marrow and/or other tissues and by the presence of serum monoclonal immunoglobulin M ([IgM], without cutoff limit). Differential diagnosis from other B-cell disorders (BCDs) is usually easy based on clinical, morphologic, histopathologic, immunophenotypic, and genetic features. However, all BCDs potentially produce monoclonal IgM. In this study we reviewed the medical files of 130 patients with IgM-secreting BCDs. Eighty-four patients were diagnosed with WM, 5 with IgM-monoclonal gammopathy of undetermined significance (MGUS), and 41 with other BCDs (9 with B-cell chronic lymphocytic leukemia, 5 with small lymphocytic lymphoma, 14 with marginal zone lymphoma, 5 with mantle cell lymphoma, 2 with follicular lymphoma, 2 with diffuse large B-cell lymphoma, 2 with cryoglobulinemia, and 2 with low-grade lymphoma not otherwise specified). Median IgM levels were 3215 mg/dL in WM, 840 mg/dL in IgM-MGUS, and 285 mg/dL in other BCDs (5 had IgM levels > 1500 mg/dL). In 10% of non-WM BCDs, monoclonal IgM was found only when more sensitive immunofixation methods were used. Forty-four percent of patients with BCDs (splenic marginal zone lymphoma or small lymphocytic lymphoma) had diagnoses that corresponded to that of WM. Careful diagnosis requires the concomitant evaluation of all parameters of BCDs together. Marginal zone lymphoma is the most frequently overlapping entity. Special attention should be given to mantle cell lymphoma in its atypical forms. Research in this field should continue to further clarify the disease entities that overlap with WM. New technology such as gene-expression profile techniques may contribute to this purpose. PMID- 15794856 TI - Immunophenotypic and cytogenetic comparison of Waldenstrom's macroglobulinemia with splenic marginal zone lymphoma. AB - Some B-cell lymphoproliferative disorders displaying a serum monoclonal immunoglobulin (Ig) M protein could be difficult to differentiate from Waldenstrom's macroglobulinemia (WM). We report on the immunophenotypic and cytogenetic characteristics of 85 patients with WM and compare them with 29 patients with splenic marginal zone lymphoma (SMZL). For immunophenotyping, WM and SMZL constantly expressed panB-cell markers (CD19, CD20, CD22, and surface Ig). However, there were differences in the k/l ratio (1.2:1 for SMZL and 4.5:1 for WM) and in some markers such as CD22 and CD11c, which were overexpressed in patients with SMZL compared with patients with WM, whereas CD25 was more frequently positive in WM (88% vs. 44%). The CD103 antigen was always negative in WM, whereas it was positive in 40% of SMZL cases. The monoclonal antibody FMC7 was usually positive in both entities: heterogeneous in WM but homogeneous in SMZL. The combination of CD25 and CD22 could differentiate between WM and SMZL. The principal molecular abnormality in WM is 6q deletion (30% in our experience), whereas in SMZL the most common abnormalities are loss of 7q (19%) along with +3q (19%) and +5q (10%). Interestingly, the incidence of IgH rearrangement was low in WM (12%) and SMZL (10%). Immunophenotypic and molecular cytogenetic studies could help to distinguish WM from SMZL. PMID- 15794857 TI - CD5, CD10, and CD23 expression in Waldenstrom's macroglobulinemia. AB - CD5, CD10, and CD23 are cell surface antigens used to distinguish B-cell disorders. The expression of these antigens and their clinical significance in Waldenstrom's macroglobulinemia (WM), an uncommon B-cell disorder, remains to be clarified. We therefore determined expression of CD5, CD10, and CD23 by flow cytometric analysis on bone marrow lymphoplasmacytic cells (CD19+ k/l light chain restricted) for 171 serially biopsied patients with findings of the consensus panel definition of WM. Importantly, we also correlated laboratory and clinical data, as well as existence of a familial history of a B-cell disorder in view of reports suggesting familial predisposition in WM. These studies demonstrated tumor cell expression of CD5, CD10, and CD23 in 15 of 171 patients (9%), 11 of 161 patients (7%), and 37 of 105 patients (35%), respectively. Coexpression of CD23 with CD5 or CD10 was common. Tumor Lymphoplasmacytic from 10 of 15 (66%) and 3 of 11 (27%) patients with WM that expressed CD5 and CD10, respectively, also showed expression of CD23 (P = 0.01 and P = 0.08, respectively). Among patients with CD23 expression, increased serum immunoglobulin (Ig) M levels were observed compared with patients without CD23 expression (P = 0.05). No differences in age at diagnosis; presence of adenopathy and/or splenomegaly; bone marrow involvement; serum IgA, IgB, and b2 macroglobulin levels; hematocrit; platelet count; or familial history of WM or a related B-cell disorder were observed among patients with and without CD5, CD10, and CD23 expression. These studies demonstrate that CD5, CD10, and CD23 are commonly found in WM and that their expression should not exclude the diagnosis of WM. Moreover, expression of CD23 may define a clinically distinct subset of patients with WM. PMID- 15794858 TI - Cell cycle analysis of Waldenstrom's macroglobulinemia. AB - Little is known about the DNA cell content and cell cycle characteristics of immunoglobulin (Ig) M monoclonal gammopathies. The autonomous clone appears to be rather heterogeneous, from mature B lymphocytes to plasma cells (PCs). We have evaluated the DNA cell content of 27 patients with IgM monoclonal gammopathies: 18 of them had Waldenstrom's macroglobulinemia (WM), and 9 were diagnosed with IgM-monoclonal gammopathy of undetermined significance (MGUS). To specifically analyze the cell cycle of the B lymphocyte and PC populations, we used a flow cytometric double-staining technique with CD19/CD20/CD22 propidium iodide for B lymphocytes and CD38/CD138 propidium iodide for PCs. In 26 of 27 patients, both subsets of tumor cells (B lymphocyte and PC) showed a diploid DNA cell content (DNA index, 1). The median percentage of proliferating B lymphocytes, S-phase + G2/M-phase, was 1.8% (range, 0.4%-4.1%). This proliferative activity was significantly lower than that observed in nonmalignant cells (5.7%; range, 0.1% 14.2%; P = 0.004) in the same sample. No differences were observed when comparing the proliferative activity of WM with that of IgM MGUS (median, 1.7% vs. 2.2%, respectively). Cell cycle characteristics of PCs were simultaneously evaluated in 9 patients, with 1.8% cells in S phase or G2/M phase. In summary, the cell cycle analysis showed that IgM monoclonal gammopathies are low-proliferative disorders, with a DNA ploidy pattern (diploid) clearly different from that of multiple myeloma. PMID- 15794860 TI - Prognostic factors and predictors of outcome of immunoglobulin M monoclonal gammopathy of undetermined significance. AB - Monoclonal gammopathy of undetermined significance (MGUS) of the immunoglobulin M (IgM) class was diagnosed at our institution in 213 patients who resided in the 11 counties of southeastern Minnesota from 1960 to 1994. The median age at diagnosis was 74 years and the median concentration of serum M-protein was 1.2 g/dL. The 213 patients were monitored for 1567 person-years (median, 6.3 years), during which 71% died. During follow-up, non-Hodgkin's lymphoma (n = 17), Waldenstrom's macroglobulinemia (n = 6), primary amyloidosis (n = 3), and chronic lymphocytic leukemia (n = 3) developed in 29 patients (14%). The number of patients with progression to lymphoid neoplasms was 15.9 times that expected in the general population. The cumulative probabilities of progression to one of these disorders were 10% at 5 years, 18% at 10 years, and 24% at 15 years. The overall average risks for progression were approximately 1.5% per year. Rates of death resulting from other diseases (cardiovascular, cerebrovascular, etc.) were 31% at 5 years, 52% at 10 years, and 65% at 15 years. Multivariate analysis revealed that only the concentration of serum M-protein at diagnosis and the serum albumin value were independent predictors of progression. It was concluded that the patients with IgM MGUS should be followed indefinitely. PMID- 15794859 TI - Potential impact of a single nucleotide polymorphism in the hyaluronan synthase 1 gene in Waldenstrom's macroglobulinemia. AB - The hyaluronan synthase 1 (HAS1) gene encodes a plasma membrane protein that synthesizes hyaluronan, an extracellular matrix molecule. Previously, in patients with Waldenstrom's macroglobulinemia (WM), we detected upregulation of HAS1 transcripts and identified aberrant splice variants of this gene. Aberrant splicing of HAS1 results from activation of cryptic splice sites. In turn, activation of cryptic donor and acceptor splice sites can be promoted by mutations occurring upstream of these sites and/or at the branch point of slicing. We measured the frequency of the HAS1 833A/G polymorphism (ie, single nucleotide polymorphism; SNP) in patients with WM and healthy donors. Additionally, HAS1 gene expression was evaluated in the same group of patients. Our observations so far suggest that HAS1 833A/G SNPs contribute to aberrant splicing of this gene; this idea is supported by the fact that 833A/G SNP is located on an exonic splicing enhancer motif. Based on the results obtained thus far, we speculate that individuals with HAS1 833G/G genotype are predisposed toward aberrant HAS1 splicing and expression of HAS1 variants, resulting in an enhanced risk of developing WM. Study of a larger group of patients and healthy donors is needed to confirm these speculations and to evaluate the prognostic significance of these findings. PMID- 15794861 TI - Clinical characteristics and outcome of immunoglobulin M-related disorders. AB - We analyzed the clinical features and prognostic factors for transformation of immunoglobulin Mrelated disorders (IgM-RDs) to malignant lymphoproliferative disease (MLD) in 83 patients with IgM-RDs. We studied 19 patients with type I cryoglobulinemias, 56 patients with type II cryoglobulinemias, 5 patients with peripheral neuropathies (PNs), and 3 patients with idiopathic thrombocytopenic purpuras. Fourteen patients with cryoglobulinemias presented with mild to moderate hepatomegaly with or without splenomegaly. Fourteen patients with type II cryoglobulinemias had arthralgias and/or vascular purpura (12 receiving corticosteroids), and 7 presented with PN. These latter patients and those with PNs without cryoglobulinemia were treated with steroids, cyclophosphamide, or polychemotherapy with/without plasma-exchange. Cumulative probability of evolution to MLD at 5 years was 15% (95% CI; 5%-25%). At a median of 62 months (12-195 months), 8 cases of IgM-RDs (8.4%) evolved to overt Waldenstrom's macroglobulinemia (n = 6), 1 case to non-Hodgkin's lymphoma, and 1 case to B-cell chronic lymphocytic leukemia. At univariate analysis, male sex (P = 0.02), IgM level > or = 3 g/dL (P < 0.0001), detectable Bence Jones proteinuria (P = 0.0005), lymphocytosis (P = 0.049), and high erythrocyte sedimentation rate (P = 0.003) significantly correlated with the evolution risk. Age, blood cell counts, b2-microglobulin level, degree of marrow lymphoplasmacytic infiltration, type of cryoglobulinemia, and hepatitis C virus positivity did not correlate with transformation. Although IgM-RDs represent a distinct clinical entity frequently requiring treatment in view of the IgM-related symptoms, their evolution probability and prognostic factors for malignant transformation seem to widely overlap those described for asymptomatic IgM monoclonal gammopathies. PMID- 15794862 TI - Prognostic factors for transformation in asymptomatic immunoglobulin m monoclonal gammopathies. AB - We evaluated the risk of transformation of asymptomatic immunoglobulin (Ig) M monoclonal gammopathy (aIgM-MG) into symptomatic lymphoproliferative disease in 384 patients, in subgroups of patients with IgM MG of undetermined significance (MGUS) and smoldering Waldenstrom's macroglobulinemia (sWM). One hundred seventy two patients with aIgM-MG with bone marrow (BM) histopathology and immunophenotyping were available for analysis. After a median of 45 months (range, 12-233 months), 45 cases of aIgM-MG (11.7%) evolved into lymphoproliferative disease, as follows: symptomatic WM (n = 41), non-Hodgkin's lymphoma (NHL; n = 2), IgM multiple myeloma (IgM-MM; n = 1), and primary amyloidosis (n = 1). Cumulative transformation probability at 5 and 10 years was 8% and 29%, respectively. The parameters significantly correlated with evolution were, at univariate analysis, BM lymphoplasmacytic infiltration, high erythrocyte sedimentation rate, hemoglobin (Hb) level, IgM size, and peripheral lymphocytosis; at multivariate analysis, the parameters were IgM size and peripheral lymphocytosis, with Hb level associated with a trend toward higher progression risk. Of the 138 cases of IgM-MGUS, 14 (10.1%) evolved (13 WM, 1 IgM MM) after a median of 75 months (range, 12-117 months); of the 34 cases of sWM, 13 (38.2%) progressed to WM after 55 months (range, 13-154 months). In patients with IgM-MGUS, event-free survival at 5 and 10 years was 95% and 83%, respectively, and in patients with sWM, 77% and 42%, respectively (P = 0.0001). Among patients with aIgM-MG, those at high risk of evolution are patients with sWM, a distinct entity with clear BM evidence of NHL. PMID- 15794863 TI - Predictive factors for response to rituximab in Waldenstrom's macroglobulinemia. AB - Rituximab is an active agent for the treatment of Waldenstrom's macroglobulinemia. However, many patients do not respond to this agent and several others develop secondary resistance. In order to identify clinical and laboratory parameters that could predict a higher likelihood for response, we evaluated 54 patients who were treated with single-agent rituximab. Twenty-four patients (44%)exhibited > or = 50% reduction of serum monoclonal protein. Previously untreated and pretreated patients had the same probability for response. Low response rates were noted in patients with serum monoclonal protein level > or = 40 g/L (17%) and serum albumin level < 35 g/L (14%). Furthermore, a multivariate analysis indicated that high serum monoclonal protein and low albumin were the dominant variables associated with shorter time to progression. The presence of 2, 1, or none of these variables was associated with median times to progression of 4 months, 11 months, and approximately 48 months, respectively. We conclude that patients with low levels of monoclonal protein and normal albumin are the best candidates for treatment with rituximab. PMID- 15794864 TI - CHOP plus rituximab therapy in Waldenstrom's macroglobulinemia. AB - Recently, a consensus panel of experts recommended that patients with Waldenstrom's macroglobulinemia (WM) who are candidates for future autologous transplantation should have limited alkylator or nucleoside analogue exposure due to potential stem cell harm. Cyclophosphamide/doxorubicin/vincristine/prednisone/rituximab (CHOP-R) is a stem cellsparing regimen that has been extensively evaluated in patients without WM or non-Hodgkin's lymphoma. As such, we analyzed the outcome of 13 patients with WM who received CHOP-R at our institution. Patients had a median age of 54 years and a median of 1 previous therapy. Ten of 13 patients (77%) had relapsed (n = 3) or refractory (n = 7) disease. Eight and 6 patients had previously received fludarabine and rituximab, respectively. Intended therapy consisted of 6 cycles of standard-dose CHOP and 6 infusions of rituximab (375 mg/m2). Three patients received additional rituximab as maintenance therapy. Median immunoglobulin M and serum viscosity for all patients decreased from 5230 mg/dL to 1690 mg/dL (P < or = 0.001) and from 2.9 cP to 1.6 cP (P = 0.01), respectively, and the median hematocrit level rose from 30.5% to 39.3% (P < or = 0.001). Clinical responses were as follows: 3 complete responses unconfirmed, 8 partial responses, 1 minor response. At a median follow-up of 9 months (range, 6 to > 37 months), 10 of the 11 patients who had a major response remained in remission. Therapy was well tolerated for most patients. Two patients had febrile neutropenia with documented bacteremia and recovered without complications. Circulating effector cell levels were also evaluated in 6 patients before and after CHOP-R, because rituximab activity is mediated in part by antibody-dependent cell-mediated cytotoxicity activity. No significant change in CD3+, CD4+, CD8+, and CD16+/CD56+ effector cell levels occurred following CHOP-R as assessed by multicolor flow cytometry. PMID- 15794865 TI - CD52 expression in Waldenstrom's macroglobulinemia: implications for alemtuzumab therapy and response assessment. AB - The efficacy of monoclonal antibody therapy is determined, at least in part, by the extent to which the target antigen is expressed. This is a complex issue in Waldenstrom's macroglobulinemia (WM) as it is a disorder characterized by plasma cell differentiation and therefore target antigen expression may differ between the B-cell and plasma cell compartments of the disease. In order to assess this in the context of alemtuzumab therapy, the authors used multiparameter flow cytometry to determine CD52 expression in the B-cells and plasma cells of patients with WM. CD52 expression was demonstrable in the B-cells of all cases, with a median of 99% of cells (range, 81%-100%) expressing the antigen compared with the isotype control (n = 47). Antigen density was very similar to that seen in chronic lymphocytic leukemia (median mean fluorescence intensity [MFI], 1249; range, 175-3170). Antigen expression was, however, significantly lower in the plasma cells (median MFI, 235; range, 31-1814) in all but 1 of the cases assessed (n = 21). The clinical significance of this was assessed by examining serial bone marrow samples from patients receiving alemtuzumab as part of an ongoing clinical trial. In 4 of 5 patients, alemtuzumab therapy successfully eradicated clonal B cells from the bone marrow, but residual plasma cells remained evident in 2 of these patients. The implications of these findings for monoclonal antibody therapy in WM are discussed. PMID- 15794866 TI - Early results of a phase I trial of oblimersen sodium for relapsed or refractory Waldenstrom's macroglobulinemia. AB - Oblimersen sodium is an antisense oligonucleotide to the first 6 codons of the B cell leukemia gene 2 (bcl-2) open reading frame. It prevents the expression of the bcl-2 gene product and leads to apoptosis in cells that express Bcl-2. bcl-2 is one of the major apoptosis regulatory gene families and is found in a variety of low-grade B-cell non-Hodgkin's lymphomas. The in vitro use of oblimersen in Waldenstrom's macroglobulinemia (WM) cell line results in enhanced toxicity when exposed to fludarabine, dexamethasone, or rituximab. Oblimersen should also enhance the cytotoxic effect of chemotherapy in WM. Presented herein are early data on the phase I portion of a phase I/II study of oblimersen in WM to identify the maximum tolerated dose and to evaluate response in patients with symptomatic WM. PMID- 15794867 TI - Immunoglobulin M myeloma: evaluation of molecular features and cytokine expression. AB - Immunoglobulin (Ig) M myeloma is a distinct entity with features of multiple myeloma (MM) and Waldenstrom's macroglobulinemia (WM). The malignant cells in IgM myeloma have a distinctive chromosomal translocation that differentiates them from WM. These cells are postgerminal-center in origin with isotype-switch transcripts. They appear to be arrested at a point of maturation between that of WM and MM. Preliminary data indicate that a pattern of osteoclast-activating factor and osteoprotegerin expression similar to that observed in classic MM is present in IgM myeloma. Additional studies on patients with this rare tumor may provide further insight into the pathogenesis of bone disease in plasma cell dyscrasias. PMID- 15794868 TI - Cold agglutinin disease and cryoglobulinemia. AB - Cold agglutinin disease is a form of direct, extravascular, antiglobulin-positive hemolysis. In vivo, immunoglobulin (Ig) M fixes complement molecules to the red cell membrane. Successive passages through the mononuclear phagocyte system result in loss of red cell membrane. The resultant spherocytes lose resiliency and are ultimately lost from the circulation extravascularly. The high concentration of complement molecules on the red cell surfaces makes this syndrome resistant to the standard therapies for immune-mediated hemolysis. Rituximab has been reported to reduce the severity of hemolysis. Type II cryoglobulins are composed of a monoclonal IgM and a polyclonal IgG. These complexes have rheumatoid factor activity and can produce immune-complex vasculitis. The target organs are the skin, nerves, kidney, liver, and joints. More than 80% of patients have evidence of hepatitis C infection. Interferon and interferon plus ribavirin have been shown to produce serologic responses. When vasculitis is active, corticosteroids are often required to permit healing of ulcers in the skin or to treat the membranoproliferative glomerulonephritis that is seen, thereby preventing loss of renal function. Rituximab therapy has been found to be effective in mixed cryoglobulinemia, with decreases in cryoglobulin values and improvement in complement values. PMID- 15794869 TI - Phase III study of chlorambucil versus fludarabine as initial therapy for Waldenstrom's macroglobulinemia and related disorders. AB - The WM1 study is a prospective randomized open-label study that includes patients with previously untreated Waldenstrom's macroglobulinemia (WM), splenic lymphoma with villous lymphocytes (SLVL), and nonimmunoglobulin (Ig) M lymphoplasmacytic lymphoma (LPL) who have an indication for treatment. At registration, patients are categorized as having WM, SLVL, or LPL, and these cohorts are to be analyzed separately. The aim of the study is to compare the efficacy of oral chlorambucil at a dose of 8 mg/m(2) (6 mg/m(2) for those > 75 years of age) for 10 days every 28 days to a maximum of 12 cycles with oral or intravenous (I.V.) fludarabine at a dose of 40 mg/m(2) orally or 25 mg/m(2) I.V. (30 mg/m(2)orally or 20 mg/m(2)I.V. for those > 75 years of age) for 5 days every 28 days to a maximum of 6 cycles. Primary endpoints are response to therapy and duration of response; secondary endpoints are improvement in hematologic parameters, toxicity of therapy, quality of life, and survival. To detect a difference in response rate of patients with WM of 15%, assuming that the overall response rates will be 50% to chlorambucil and 65% to fludarabine, with a power of 80%, requires the sample size of each group to be 183, indicating the need for collaboration among a number of national investigator groups. As of February 2005, accrual to the study stands at 143. Registration, randomization, and data collection are entirely Internet-based (www.waldenstroms.org), and the study is organized by an international collaboration, with a planned interim analysis and an external data monitoring committee. PMID- 15794870 TI - Strategies of international cooperation in an international project: advantages and pitfalls. AB - The MEPS (musculoskeletal--eyestrain--psychosocial--stress) study involved an extensive degree of multidisciplinary and multicultural cooperation. The objective was to examine the effects of multiple ergonomic interventions on visual, musculoskeletal, postural, and psychosocial outcomes amongst operators of visual display terminals (VDTs). The inherent complexity of a comprehensive ergonomic investigation requires participation of researchers from a variety of disciplines, as well as comparisons among populations with different geographical and cultural backgrounds. The design and execution of the resulting research protocol presents a number of challenges. This paper discusses the advantages and pitfalls associated with multidisciplinary multinational cooperation. Advantages include the necessity for development of a common language and perspective providing a basis for future collaboration. Pitfalls include logistic and coordination difficulties associated with conducting standardized procedures in different locations, as well as the inherent potential for professional conflict. It is argued that such pitfalls ought to be understood and integrated into the project planning process. PMID- 15794872 TI - The effect of an ergonomic intervention on musculoskeletal, psychosocial and visual strain of VDT data entry work: the Norwegian part of the international study. AB - The Norwegian MEPS (musculoskeletal--visual--psychosocial--stress) study included 3 groups: data entry, data dialogue (female) and data dialogue (male). Before intervention, the data entry group reported significantly more symptoms and signs of musculoskeletal illness and had longer periods in front of the video display terminal (VDT) without a break. The ergonomic intervention consisted mainly of ergonomic information and training. After intervention, the data dialogue female group reported a significant reduction in shoulder pain in parallel with a reduction in trapezius load. Increasing the understanding in how to adjust the work stand and chair may have been contributing factors to reducing the pain level. There was a significant reduction in eye problems in all groups; the greatest reduction in eye symptoms was seen in the groups who had new optometric corrections. PMID- 15794871 TI - The effect of an ergonomic intervention on musculoskeletal, psychosocial and visual strain of VDT entry work: organization and methodology of the international study. AB - This special issue of the International Journal of Occupational Safety and Ergonomics (JOSE) reports the results from an extensive multinational and multidisciplinary collaborative investigation of the impacts on visual display terminal (VDT) work of musculoskeletal, visual, ergonomic, and psychosocial factors. For brevity, this effort has been referred to as the MEPS project (musculoskeletal--eyestrain--psychosocial--stress). This paper lays out the basic methodological structure of the study. The study was conducted in 4 countries utilizing VDT data entry workers as the primary subject population. A battery of objective and subject assessment measures, including muscle load, visual function, physical and visual strain, postural, ergonomic and psychosocial factors, were assessed at 3 different points in time. A pre-test was given prior to an ergonomic intervention. Two post-tests were given 1 month and 1 year after the ergonomic intervention. PMID- 15794873 TI - The effect of an ergonomic intervention on musculoskeletal, psychosocial, and visual strain of VDT data entry work: the United States part of the international study. AB - The United States MEPS (musculoskeletal--eyestrain--psychosocial--stress) study consisted of 1 group of 28 female data entry operators. The intervention was in 3 parts: workstation redesign (including advanced ergonomic chairs, motorized adjustable workstations, advanced adjustable keyboards, adjustable copyholders, adjustable footrests, monitor support surfaces) and ergonomic training/coaching and corrective lenses. After the intervention, statistically significant reductions in physical signs (trigger points, neck and shoulder mobility), subjective reports of intensity and frequency of musculoskeletal pain, and subjective reports of visual problems were observed. Static load during the work sample, as assessed by experts, improved after the intervention as did measured postural angles of head and trunk and subjective assessment of users of ergonomic characteristics of the workplaces. For all of these measures, improvements observed 1 month after intervention were also observed in the 1-year follow-up. Trapezius load, as assessed by electromyography (EMG), decreased after intervention, but then increased in the follow-up. The increase was interpreted as a calibration problem. PMID- 15794874 TI - The effect of an ergonomic intervention on musculoskeletal, psychosocial, and visual strain of VDT data entry work: the Polish part of the international study. AB - A group of routine data entry operators (female) was included in the Polish MEPS (musculoskeletal--eyestrain--psychosocial--stress) study. Before the intervention, ergonomic assessment revealed improper working conditions such as inadequate lighting, uncomfortable chairs, and lack of forearm and wrist support while medical examination revealed that trapezius muscle load along with upper arm, head and back angles were higher than advisable. Subjects complained about neck and shoulder pain, visual problems, and psychosocial conditions. The ergonomic intervention included installation of new luminaires and Venetian blinds, new chairs, repair of ventilators, and optometric corrections. The results after the intervention showed mainly improvement in chair comfort, lighting conditions, visual strain, and sitting posture. However, financial limitations did not allow satisfactory completion of the intervention leading to a mixed interpretation of the results. PMID- 15794876 TI - Upper limb load as a function of repetitive task parameters: part 1--a model of upper limb load. AB - The aim of the study was to develop a theoretical indicator of upper limb musculoskeletal load based on repetitive task parameters. As such the dimensionless parameter, Integrated Cycle Load (ICL) was accepted. It expresses upper limb load which occurs during 1 cycle. The indicator is based on a model of a repetitive task, which consists of a model of the upper limb, a model of basic types of upper limb forces and a model of parameters of a repetitive task such as length of the cycle, length of periods of the cycle and external force exerted during each of the periods of the cycle. Calculations of the ICL parameter were performed for 12 different variants of external load characterised by different values of repetitive task parameters. A comparison of ICL, which expresses external load with a physiological indicator of upper limb load, is presented in Part 2 of the paper. PMID- 15794875 TI - A cross-country comparison of short- and long-term effects of an ergonomic intervention on musculoskeletal discomfort, eyestrain and psychosocial stress in VDT operators: selected aspects of the international project. AB - Three groups of data entry female visual display terminal (VDT) workers from Norway (n = 30), Poland (n = 33) and the USA (n = 29) were compared. Before intervention, the Norwegian group reported more neck pain compared with the Polish group. The Polish group reported less shoulder pain than both the U.S. and the Norwegian groups. The clinical examination documented fewer symptoms and signs of musculoskeletal illness among the Polish participants compared with the Norwegian and the U.S. groups. After intervention, the Norwegian group reported a reduction in neck pain while the U.S. group reported a reduction in shoulder pain. The Polish group reported an increase in neck, shoulder and forearm pain at follow-up compared to after intervention. The Polish group recorded higher flexion of the upper arm at follow-up parallel with an increase of pain in the upper part of the body. Visual discomfort showed variable results in the 3 countries. PMID- 15794877 TI - Upper limb load as a function of repetitive task parameters: part 2--an experimental study. AB - The aim of the study was to compare the theoretical indicator of upper limb load with the physiological indicator of musculoskeletal load, which is present while performing a repetitive task (a normalized electromyography [EMG] amplitude recorded from the muscles of the upper limb involved in the performed task). In an experimental study of a repetitive task, the EMG signal from 5 main muscles of the shoulder girdle, arm and forearm was registered: extensor carpi radialis longus, flexor carpi ulnaris, deltoideus anterior, biceps brachii caput breve and trapezius descendent. The results of the study showed a strong correlation between the theoretical indicator (Integrated Cycle Load) and the physiological indicator (root mean square of a normalized EMG amplitude from the 5 muscles). This proves that the developed theoretical indicator can be accepted as an indicator of upper limb musculoskeletal load during a work task. PMID- 15794879 TI - [Control and prevention of cardiovascular diseases]. PMID- 15794881 TI - [Prevalence of isolated systolic hypertension in Chinese adult population]. AB - OBJECTIVE: To evaluate prevalence of isolated systolic hypertension and other subtypes of hypertension, as well as their distribution in Chinese adults. METHODS: A cross-sectional survey was conducted in the mainland China during 2000 2001 with a multistage cluster sampling to select a nationally representative sample of 15,540 Chinese adults aged 35-74, by InterASIA project. Hypertension subtypes were defined among individuals not receiving antihypertensive therapy as follows: isolated systolic hypertension as a systolic blood pressure > or = 140 mm Hg and a diastolic blood pressure < 90 mm Hg, isolated diastolic hypertension as a systolic pressure < 140 mm Hg and a diastolic pressure > or = 90 mm Hg, and combined systolic/diastolic hypertension as a systolic pressure > or = 140 mm Hg and a diastolic pressure > or = 90 mm Hg. Blood pressure for each subject was measured for three readings at sitting position after rest for at least five minutes and an average reading was recorded. A standardized structured questionnaire was used to collect history of hypertension and antihypertensive treatment. RESULTS: Overall, 7.6% (1,181 cases) of adults had isolated systolic hypertension, 7.4% (1,150 cases) combined systolic/diastolic hypertension and 4.4% (683 cases) isolated diastolic hypertension in the sample of 15,540 adults at age 35-74. Prevalence of systolic hypertension increased with age, and was more common in the older women than in the older men. There was no significant difference in prevalence of isolated systolic hypertension between north and south China, but, rural residents had a higher prevalence than urban residents. CONCLUSION: There existed a higher prevalence of systolic hypertension in China, to which enough attention should be paid. PMID- 15794882 TI - [Changes of body mass index in a community cohort of Peking University from 1992 to 2002]. AB - OBJECTIVES: To compare changes in body mass index (BMI), prevalence of over weight and obesity of the population in a community cohort of Peking University during 1992 to 2002, and to study their characteristics of individual BMI. METHODS: Totally, 1 985 individuals aged 35-64 years were selected from Peking University community with stratified random sampling method for baseline survey by physical examinations and questionnaire interviews to collect relevant data in 1992. And, a follow-up survey was conducted for each individual of the cohort in 2002. Data were analyzed for 1 437 individuals with complete records of the two surveys. RESULTS: (1) In the period from 1992 to 2002, weight increased averagely 4.89 kg in men and 3.08 kg in women, respectively, and BMI increased to 25.22 kg/m2 in 2002 from 23.49 kg/m2 ten years ago in men and to 25.02 kg/m2 from 23.30 kg/m2 in women. (2) Prevalence of overweight increased by 47.8% in men and 29.5% in women, and prevalence of obesity increased by 124.2% in men and 131.3% in women, respectively. (3) 74.9% of men with overweight ten years ago remained overweight and 22.9% of them became obese in 2002. 85.7% of men with obesity ten years ago remained obese in 2002. 58.2% of women with overweight ten years ago remained overweight and 34.5% of them became obese in 2002. 93.7% of women with obesity ten years ago remained obese in 2002. CONCLUSIONS: Mean BMI and prevalence of overweight and obesity of population in Peking University community increased during 1992 to 2002. Most of those with overweight or obesity ten years ago remained overweight or obese and few of them became normally weighed ten years later. Those with overweight BMI were more liable to become obese than those with normal BMI. All these results indicated once overweight or obesity established, it was difficult to be reversed. Therefore, the key to control overweight and obesity rests on its earlier prevention. PMID- 15794883 TI - [Relationship of the HSG gene single nucleotide polymorphism and hypertension]. AB - OBJECTIVE: To investigate the single nucleotide polymorphism (SNP) of the novel hyperplasia suppressor gene (HSG) to uncover the relationship between HSG SNP and hypertension. METHODS: Totally, 74 normotensive people (38 men and 36 women), 51 patients with essential hypertension (27 men and 24 women) and 20 hypertensive patients with family history of essential hypertension (9 men and 11 women) were chosen, with mean ages of (54 +/- 8) years, (57 +/- 8) years and (38 +/- 22) years, respectively. Peripheral venous blood specimen was collected from each of them and then DNA was extracted. The right primers were designed for DNA amplification with PCR. Each of the PCR-products from different groups was sequenced by ABI PRISM 377-DNA sequencer and their base components and characteristics of the same fragment were compared each other. RESULTS: Blood levels of creatinine (CRE) and urea nitrogen (BUN) were significantly higher in the hypertensives than in the normotensives (P < 0.01). Systolic and diastolic blood pressures were significantly higher in the hypertensives and the hypertensives with family history of essential hypertension than in the normotensives (P < 0.01 and P < 0.05). There existed three kinds of SNP in the HSG 12th intron (1q82139 G/A, 82153C/G and 82273G/-), and there was significant difference in 1q82153C/G and 82273G/- SNP between the hypertensives and normotensives (P < 0.05 and P < 0.01) and between the hypertensives with family history and the normotensives (P < 0.01 and P < 0.01). And, the similar difference in C/G allele and G deletion could be found in different populations. Moreover, the CC genetype of 1q82153 was common in the population (P < 0.01) and G deletion was more common in Chinese Han people with family history of essential hypertension. There was no significant difference existed in 1q82139 G/A mutation among the three groups. CONCLUSIONS: Measurements of renal function indicators (CRE and BUN) could probably reflect earlier advance of hypertension and damage to target organs. There existed three kinds of mutation in the 12th intron of the HSG 1q82139 G/A, 82153 C/G and 82273 G/-. The 1q82139 G/A could be a nonsense mutation and there was significant difference in the 1q82153 C/G and 1q82273 G/- SNP and gene frequencies between different Chinese Han populations, which could be independent risk factors for essential hypertension. PMID- 15794885 TI - [Neuroprotection by resveratrol against cytotoxicity of oxidized low density lipoprotein in PC12 cells]. AB - OBJECTIVE: To investigate neuroprotection of resveratrol against cytotoxicity of oxidized low density lipoprotein (oxLDL) in PC12 cells. METHODS: PC12 cells were pretreated with resveratrol for one hour as a neuron model and then exposed to oxLDL at varied concentrations. Effects of resveratrol on cell viability, permeability of cell membrane, cell nucleus and activity of caspase-3 were evaluated with MTT assay, lactic acid dehydrogenase (LDH) release assay, DNA fragmentation (TUNEL) assay and assay for caspase-3 activity (caspase assay). RESULTS: Cell viability, LDH release rate, percentage of cells with positive TUNEL and activity of caspase-3 were (62 +/- 3)%, (23 +/- 3)%, (26 +/- 5)% and (0.811 +/- 0.049) mol.min(-1).microg(-1), vs (84 +/- 7)%, (13 +/- 4)%, (12 +/- 4)% and (0.553 +/- 0.048) mol.min(-1).microg(-1) in PC12 cells treated with 10 mg/L oxLDL vs in those treated with 10 mg/L oxLDL plus 50 micromol/L resveratrol, respectively, with statistically significant difference. CONCLUSION: Resveratrol could attenuate cytotoxicity induced by oxLDL in PC12 cells with neuroprotection. PMID- 15794886 TI - [The investigation on basic contents of organochlorine pesticides in human milk during consecutive periods in Beijing urban areas]. AB - OBJECTIVE: To investigate the basic contents of organochlorine pesticides (DDT, BHC and HCB) in human milk of normal residents in Beijing urban areas. METHODS: Through continuous survey on the trend of organochlorine contents in human milk since DDT and BHC pesticides were forbidden in use in 1983, and to evaluate the effects of this program, the required lying-in women residents in urban districts for at least 5 years were enrolled in this study. All these lying-in women and also their family members were not exposed to or engaged in producing or selling organochlorine pesticides. In this study, the women chosen were giving birth to a child just after one week to 4 months. The detection method used is the national standard biological detection method. RESULTS: In 1982, the total amount of DDT and BHC contents in human milk were 6.45 mg/kg and 6.97 mg/kg, respectively, which were found the highest level in the word. The contents of organochlorine were descending apparently since the DDT and BHC pesticides were forbidden by Chinese government in 1983. According to the standards of FAO/WHO, the average intake of total DDT and beta-BHC in the infants born in 1998 was 11 and 6 microg/kg, respectively. Both of them were less than the allowable daily intake amount regulated by WHO and the Ministry of Health of Japan. CONCLUSION: The study might make clear that the basic levels of organochlorine pesticides (DDT, BHC and HCB) in human milk of normal residents and the attenuation trend of 2 pesticides cumulative levels in human body, through estimating the intake quantity of organochlorine by infants, showing that the DDT and BHC daily intake from human milk absorbed by infants, who were born after 1998, were at a safe level. It was indicated that the measure for forbidding use of DDT and BHC pesticides is contributory. PMID- 15794887 TI - [Fluorosis on expression of nicotinic acetylcholine receptors in protein and gene levels in human SH-SY5Y neuroblastoma cells]. AB - OBJECTIVE: To investigate the influence of fluorosis on nicotinic acetylcholine receptors (nAChRs) in protein and gene levels in SH-SY5Y cells and the mechanism of the receptor modification. METHODS: SH-SY5Y cells, a human neuroblastoma cell line, were incubated with different concentrations of fluoride or with antioxidant for 48 hours. The functions of cells were measured by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) method, and protein oxidation detected by carbonyl content; the alpha3 and alpha7 nAChR subunits in protein level were measured by Western blotting and in mRNA level by RT polymerase chain reaction (RT-PCR). RESULTS: In high-dose group as compared to the control, the decreased MTT (49%), increased protein oxidation (72%), and lower expression of alpha3 (51%) and alpha7 (47%) nAChR subunit proteins were obviously observed in SH-SY5Y cells. There were no changes in expression of nAChR subunit mRNAs between the cells treated with fluoride and those un-treated in controls. Prior treatment with antioxidant resulted in preventing the decrease of nAChR protein in cells exposed to the high doses of fluoride. CONCLUSION: Fluorosis should result in damage of cells and the declined expression of nAChRs in protein levels, but no influences on gene expression of the receptors in human neuroblastoma neurons. The decreased nAChR proteins might be involved in the mechanism of oxidative stress induced by fluorosis. PMID- 15794888 TI - [Effects of various iodin-nutritional on activity of T4 5'-and 5-deiodinase in rat brain]. AB - OBJECTIVE: To investigate the changing of T4 5'-and 5-deiodinase within rat brain under various iodin-nutritional states. METHODS: Animal model of iodine deficiency rat was performed and the rats were divided into 4 groups by the intake of iodine-nutrition, and then killed at an age of 20 days. The thyroid hormones level in serum was measured by ELISA and the activity of T(4) 5'-and 5 deiodinase within brain was analyzed. RESULTS: In less-iodine (LI) group,TT4 and FT4 were accounting for 3.5% of the neutral-iodine (NI) group's, and FT3 was 174.0% of NI group's (P < 0.05). In NI group,TT4 and FT4 were 114.5% and 127.7% of NI group's (P < 0.05). In high-iodine (HI) group, TT4 and FT4 were 61.86% and 62.0% of NI group's, and FT3 was 184.9% of NI group's (P < 0.05). In LI group, the activity of T4 5'-deiodinase tissue of per gram (1.95 +/- 0.32) ngT3.microgT4(-1).h was significantly higher than that of NI group (P < 0.05), and the activity of 5-deiodinase (1.38 +/- 0.21) ngrT3.microg T4(-1).h(-1) is significantly less than that of NI group (1.59 +/- 0.23) (P < 0.05). In HI group the activity of T4 5'-and 5-deiodinase tissue of per gram (1.12 +/- 0.19 and 1.73 +/- 0.36) ngrT3.microgT4(-1).h(-1)was significantly less than that of NI group (P < 0.05). CONCLUSION: The activity of T4 5'-deiodinase in iodine deficiency heightens and that in iodine excess is debased, the activity of T4 5-deiodinase in iodine deficiency and in iodine excess is debased. PMID- 15794889 TI - [Effect of conjugated linoleic acid on gene expression of adiponectin of obese rat fed with high fat diet]. AB - OBJECTIVE: To study the effect of conjugated linoleic acid (CLA) on expression of adiponectin in white adipose tissue of obese rats. METHODS: Male Wistar rats were randomly divided into control group, high-fat group and high fat + CLA group (0.75 g, 1.50 g, 3.00 g per hundred gram diet weight), we observed the effect of CLA on serum insulin and glucose levels of obese rats, and the reverse transcription polymerase chain reaction (RT-PCR) technique was used to measure the expression level of adiponectin and peroxisome proliferator-activated receptor-gamma (PPARgamma) mRNA. RESULTS: The serum insulin and glucose levels of obese rats were (11.11 +/- 2.73) microIU/ml, (5.09 +/- 0.66) mmol/L. The supplement of CLA decreased the hyperinsulinemia and hyperglycemia, the serum insulin in CLA group (0.75 g, 1.50 g, 3.00 g per hundred gram diet weight) were (6.99 +/- 1.77) microIU/ml, (7.36 +/- 1.48) microIU/ml, (7.85 +/- 1.60) microIU/ml (P < 0.05), and glucose were (4.28 +/- 0.72) mmol/L, (4.18 +/- 0.55) mmol/L (P < 0.05), (4.06 +/- 0.63) mmol/L (P < 0.05), CLA can increase the expression of adiponectin and PPARgamma in adipose tissue of obese rat. CONCLUSION: The CLA might improve the insulin resistance of the obese rat and increase the expression of adiponectin mRNA, which might possibly act through activating PPARgamma. PMID- 15794890 TI - [Investigation of acquired immunodeficiency syndrome correlated with high risk sexual behavior and knowledge of male homosexuality in Hangzhou Province, China]. AB - OBJECTIVE: To find out the acquired immunodeficiency syndrome (AIDS) correlated high risk sexual behavior and knowledge in male homosexuals in Hangzhou Province. METHODS: Using an self-designed anonymous questionnaire, 927 male homosexual were investigated anonymously in May 2003 to January 2004. RESULTS: Among the male homosexuals surveyed, the average age of this population was (29.8 +/- 9.9) years, the average number of sexual partners per person in past one year was 7.34 +/- 5.61. Among them, 199 (21.46%) had one sexual partner, 165 (17.79%) had 2 - 5 sexual partners, and 563 (60.73%) had more than 5 sexual partners, and 179 (19.30%) also had opposite-sex sexual partner the average number of opposite-sex sexual partner of this group was 1.16 +/- 0.21. According to sexual behavior, 40.66% of them used anal sexual behavior, 62.99% used oral sex, 74.11% used masturbation and 3.66% used oral-anal touch. Among 179 male homosexuals having opposite sexual partners, the percentage of using vagina sex, anal sex, oral sex and masturbation was respectively 59.77%, 36.87%, 40.22% and 26.25%. Although their understanding level about AIDS was high, the risk rate of sexual behavior among them was also very high. Only 11.11% of male homosexuals used condom when they had anal sex with same sex, most of them rarely used or never used protection measures. When having sex with female, almost none of them used protection. CONCLUSION: The risk rate of sexual behaviors in male homosexuals is very high that some appropriating and effective measures for prevention should be taken. PMID- 15794891 TI - [Effects of nucleotides on apoptosis of thymocytes]. AB - OBJECTIVE: To study the effects of nucleotides on apoptosis of thymocytes in mice. METHODS: Apoptosis model in vivo was first established and 25 KM mice, 4 weeks old, were randomly divided into 5 groups. One group was control, and the others were test groups. Mice in test groups were injected with DEX (25 mg/kg) and the controls were treated with normal saline. 4, 8, 16, and 24 hours later the thymus and spleen were weighed and lymphocytes in thymus were separated. The apoptosis of lymphocytes was analyzed by using DNA electrophoresis and flow cytometry. 16 hours later lymphocytes apoptosis reached a peak and lasted 24 hours. Methods used to establish apoptosis model in vivo were: mice (4 weeks old) were injected with DEX (25 mg/kg), and thymus lymphocytes were separated 16 hours later and analyzed. The effects of nucleotides on apoptosis of mice thymocytes were investigated in experiment 2. Sixty KM mice, 20 g +/- 2g, 4 weeks old, were divided into four treatments: negative control group (NC), positive control group (PC), nucleotides-additive group 1 (NTS1) and nucleotides-additive group 2 (NTS2). RESULTS: Body weight gained in NST1 and NST2 were 3.71 g, 4.01 g respectively, significantly higher than NC (2.74 g) (P < 0.01) and in NST2 was significantly higher than in PC (2.96 g) (P < 0.01). Thymus index and spleen index were decreased significantly (P < 0.01), and no difference was found with the supplementation of nucleotides (P > 0.05). [Ca2+]i increased to 167.37 nmol/L, 191.16 nmol/L, 180.78 nmol/L in PC, NST1 and NST2 with DEX, being significantly higher than in NC (103.76 nmol/L) (P < 0.01). The percent of apoptosised thymocytes in groups were 0.31%, 11.93%, 9.82%, 11.15%, respectively. Thymus index and spleen index, cell apoptosis and [Ca2+]i were not differed significantly among PC, NTS1 and NTS2 groups. CONCLUSION: Nucleotides should have no significant effects on apoptosis of thymocytes in mice in vivo. PMID- 15794892 TI - [Cross-sectional study on falls in residents of four communities in Zhejiang Province, China]. AB - OBJECTIVE: To understand the incidence of falls in different residents of four communities in Zhejiang Province to provide a basis for their prevention and control. METHODS: A household questionnaire survey on falls was conducted in 16 899 residents of two urban and two rural communities during November 2000 to October 2001. RESULTS: Standardized incidence rate of falls was 5.07%, higher in rural areas (5.23%) than that in urban areas (4.95%) and showing no significant differences by gender. Falls occurred in September and October accounted for 11.68% and 19.22% and those occurred in pre-school children and the elderly accounted for 22.94% and 19.46% of the total episodes, respectively. Leading causes of falls varied in different population, from falls during their play (7.65%) and sports (4.06%) in young people and pre-school children (19.12%), slipping down when walking in adult (1.21%) and elderly (4.28%) men, and slipping down when up and down stairs in adult and elderly women. Burden and severity of injury caused by falls were increased with age. CONCLUSIONS: Incidence of falls in pre-school children and the elderly was higher in local residents of communities in Zhejiang Province, causing heavy burden to the society and their families. Varied relevant measures should be taken to prevent and control for falls in different subgroups of population. PMID- 15794907 TI - In vitro and in vivo approaches to prevention and cure of T1D. PMID- 15794893 TI - [Investigation on sleep status of college and high school students]. AB - OBJECTIVE: To investigate the sleep status of college and high schools students. METHODS: Pittsburgh sleep quality index (PSQI) and self-manufactured questionnaires about siesta habits were used as tools. Three groups of students from medical college (MC), senior high school (SS) and junior high school (JS) were surveyed. RESULTS: In the group MC, SS and JS, the occurrence rates of sleep disorders were 27%, 62% and 54%, respectively, and in which the appearance rates of insomnia were 17%, 19% and 19%, longing for sleep were 10%, 43% and 35% respectively. And there were no significant difference between schoolboy and schoolgirl. The occurrence rates of slack breathing were different (5/155, 1/154) significantly between group SS and JS. The distinct differences also were found in group JS and MC, in which students felt hot (10/155, 1/122) and in all the three groups, in which students felt sleepy (55/155, 62/154, 13/122) whereas the difference of sleepy between group SS and JS was comparatively distinct (55/155, 62/154). Significant differences were also found between group JS and SS, MC in average sleep time of (7.65 +/- 0.87) hours, (7.16 +/- 0.83) hours, and (7.10 +/- 0.57) hours. The time of falling asleep (median 15 min, 10 min, 20 min) and siesta habit (8/155, 19/154, 75/122) among group MC and SS, JS were different respectively and markedly, whereas siesta habit differences between group SS and JS were comparatively distinct (8/155, 19/154). CONCLUSION: Students in high school showed higher rate of longing for sleep, and this implicated they fall short of sleep time greatly and siesta could improve their sleepy signs. PMID- 15794908 TI - Statin treatment of patients with diabetes. PMID- 15794909 TI - Polyendocrine autoimmunity. AB - Autoimmune polyendocrine disorders are characterized by the presence of more than one autoimmune endocrine disease. Study of such disorders has provided unique insight into the pathogenesis of autoimmunity and the development of tolerance in the normal individual. Autoimmune polyendocrine syndromes I and II have distinctive inheritance patterns, genetic causes, component diseases, and implications for follow-up. Care for individuals with these diseases requires monitoring for additional autoimmune disorders and aggressive treatment of identified diseases to prevent morbidity and mortality in affected individuals. PMID- 15794910 TI - Introduction of hyperglycemia and dyslipidemia in the pathogenesis of diabetic vascular complications. AB - Hyperglycemia and dyslipidemia are significant and independent risk factors for the vascular complications in patients with diabetes. They have been suggested to cause cardiovascular pathologic changes in diabetic states through the following molecular mechanisms: formation and accumulation of advanced glycation end products; increased oxidative stress; activation of protein kinase C pathway; increased activity of hexosamine pathway; and vascular inflammation and the impairment of insulin action in the vascular tissues. PMID- 15794913 TI - Tackling the hard problems: long-term maintenance. PMID- 15794911 TI - Predicting type 1 diabetes. AB - Predicting type 1 diabetes mellitus (T1DM) is a prerequisite for disease prevention. Prediction is currently performed on three levels, which include the genetic susceptibility for disease, the identification of preclinical T1DM by way of circulating islet autoantibodies, and the use of metabolic tests to stage preclinical disease into late or early prediabetes. Combinations of genetic markers such as HLA genotype, INS genotype, and if and how much family history of T1DM is present can stratify disease risk more than 1000-fold, and can be used for selection of first-degree relatives of patients with T1DM for primary intervention trials. Measurement of autoantibodies in genetically at-risk subjects identifies future cases of T1DM. Further stratification of diabetes risk in autoantibody-positive subjects can be made on the basis of autoantibody characteristics that correspond to the magnitude of the autoantibody response. PMID- 15794912 TI - Regulatory T cells and type 1 diabetes. AB - Regulatory T cells are now recognized as important mediators of self-tolerance and may mediate responses to immune therapy. The mechanisms of action of these cells are diverse, and some studies suggest that there may be defects in regulatory cells in patients with type 1 diabetes. These cells may be expanded by immune therapy, suggesting the possible development of adoptive immune therapy to transfer regulation with the cells. PMID- 15794914 TI - Tackling the hard problems: intensity of intervention and reimbursement issues. PMID- 15794917 TI - Does bariatric surgery reduce obesity-related comorbidities? PMID- 15794915 TI - Patient-reported assessments in diabetes care: clinical and research applications. AB - Patient-reported assessments play an increasing role in diabetes care and research today. These assessments are used to evaluate new treatment regimens, explore attitudes, beliefs, and needs of diabetes populations, support inclusion of psychosocial aspects in daily care, and establish new patient-centered standards for quality-of-care improvement initiatives. Much work remains to be done to ensure that these activities will result in improved access to patient centered diabetes care. PMID- 15794916 TI - Health-related quality of life among adults with diabetes. AB - Diabetes has significant effects on quality of life; however, the interrelationships are complex among the physiologic and psychosocial effects of diabetes, the effects of treatment regimens, and the social and cultural contexts in which we live. These important relationships and effects are being defined, but much additional work is needed to reliably and accurately measure health related quality of life among subpopulations, and to measure and optimize the effects of increasingly complex and intensive treatments. PMID- 15794918 TI - Adiponectin: an adipokine linking adipocytes and type 2 diabetes in humans. AB - Adipocyte-derived adiponectin is an insulin-sensitizing and antiatherosclerotic hormone, and replenishment of adiponectin in animal models ameliorated insulin resistance and atherosclerosis. In humans, recent studies have demonstrated that adiponectin level is a good predictor of developing type 2 diabetes and coronary artery disease. Decreasing level of adiponectin is caused by the interaction between genetic factors, such as single nucleotide polymorphisms in the adiponectin gene, and environmental factors, such as high-fat diet. Agents that increase blood level of adiponectin or enhance the actions of adiponectin can be an ideal medicine for ameliorating insulin resistance and type 2 diabetes. PMID- 15794919 TI - Nonobese diabetic mice and the genetics of diabetes susceptibility. AB - The nonobese diabetic mouse spontaneously develops an autoimmune, T-cell-mediated type 1 diabetes (T1D). Common and rare alleles both within a diabetogenic major histocompatibility complex (MHC) and multiple non-MHC genes combine to impair normal communication between the innate and acquired immune system, leading to loss of immune tolerance. An understanding of how variable collections of genes interact with each other and with environmental cues offers important insights as to the complexities of T1D inheritance in humans. PMID- 15794920 TI - The role of HNF4A variants in the risk of type 2 diabetes. AB - Genes influence susceptibility to type 2 diabetes mellitus (T2DM), and both positional cloning and candidate gene approaches have been used to identify these genes. Linkage analysis has generated evidence for T2DM-predisposing variants on chromosome 20q in studies of Caucasians, Asians, and Africans, and fine-mapping recently identified a likely susceptibility gene, hepatocyte nuclear factor 4 alpha (HNF4A). Rare loss-of-function mutations in HNF4A cause maturity-onset diabetes of the young and now common noncoding variants have been found to be associated with T2DM. PMID- 15794922 TI - A hypomorphic allele of the first N-glycosylation gene, ALG7, causes mitochondrial defects in yeast. AB - The modification of proteins at asparagine residues with oligosaccharides (N glycans) plays critical roles in diverse cell functions. N-glycans originate from a common lipid-linked oligosaccharide (LLO) precursor whose synthesis is initiated by the Dol-P-dependent GlcNAc-1-P transferase (GPT) encoded by an essential ALG7 gene. To identify cellular processes affected by ALG7 and N glycosylation, we replaced the genomic copy of ALG7 with its hypomorphic allele in two genetically distinct haploid yeast cells. We show that ALG7 knockdown gave rise to an unexpected phenotype of mitochondrial dysfunction. The alg7 mutants did not grow on glycerol and DNA arrays revealed the absence of mitochondrial genes' expression. Accordingly, the alg7 mutants displayed no detectable mtDNA and respiratory activity. Both mutants exhibited diminished abundance of LLO and under-glycosylation of carboxypeptidase Y (CPY). Moreover, another N glycosylation mutant with a LLO defect, alg6, was respiratory deficient. Collectively, our studies provide evidence that the dysregulation of N glycosylation in haploid yeast cells leads to mitochondrial dysfunction. PMID- 15794921 TI - Genes of the thymidine salvage pathway: thymine-7-hydroxylase from a Rhodotorula glutinis cDNA library and iso-orotate decarboxylase from Neurospora crassa. AB - Genes for two enzymes in the thymidine salvage pathway, thymine-7-hydroxylase (THase; official name thymine dioxygenase) and iso-orotate decarboxylase (IDCase) have been isolated from fungal sources. THase was isolated from a Rhodotorula glutinis cDNA library using a degenerate oligonucleotide based on the published amino acid sequence. The coding sequence was transferred to an Escherichia coli expression system, from which recombinant THase activity was measured using 14C labeled thymine. The THase sequence shows an almost complete avoidance of codons ending in A or T: 95.8% GC content is present in the third position of codons. A connection between this codon bias and the role of the thymidine salvage pathway in pyrimidine metabolism is proposed. The THase sequence is similar to Group I Fe+2-dependent, alphaKG-dependent dioxygenases. The R. glutinis THase gene was used to locate the probable THase genes in the sequenced genomes of Neurospora crassa and Aspergillus nidulans. The genes neighboring THase in these two genomes are similar to each other, and are similar to the mammalian 2-amino-3 carboxymuconate-6-semialdhyde decarboxylase (ACMSD), leading to their identification as IDCase genes. The N. crassa version was isolated by PCR of genomic DNA, and IDCase activity was measured in recombinant E. coli carrying this gene. A new family of decarboxylases, using similar substrates, is identified by virtue of the protein sequence similarity. PMID- 15794923 TI - A twin-arginine translocation (Tat)-mediated phage display system. AB - The major limitation of conventional phage display is caused by its dependence on the Sec translocation pathway. All proteins displayed on filamentous phages must first be transported into the bacterial periplasm in an unfolded state via the Sec translocation machinery. Proteins that require a cytoplasmic environment and/or cytoplasmic components for folding, or that contain "stop transfer" signals, or reach their native state before they interact with the Sec proteins are not compatible with the Sec pathway. They can never be presented using conventional phage display. We have developed an alternative phage display system, termed the TPD system, which overcomes these limitations of conventional phage display by exploiting the properties of the twin-arginine translocation (Tat) pathway. The Tat pathway only exports folded proteins that have already attained their native conformation in the cytoplasm. We investigated the functional efficiency of the TPD system by displaying and panning for a mutant of the green fluorescent protein. PMID- 15794925 TI - Osteogenic and chondrogenic differentiation of embryonic stem cells in response to specific growth factors. AB - Reliable in vitro conversion of pluripotent embryonic stem (ES) cells into bone and cartilage-forming cells would expand opportunities for experimental investigations of skeletogenesis and could also provide new cellular sources for pharmaceutical screening and for cell therapy applications. Here, we evaluate the generation of mesenchymal cell lineages from mouse ES cells following treatment of embryoid bodies with retinoic acid, previously reported to induce development of adipocyte precursors. We find that retinoic acid reduces mesodermal differentiation but enhances expression of markers of neural crest, an alternative origin of mesenchymal elements. Runx1 and Ptprv appear to provide early markers of mesenchymal potential. Subsequently, different mesenchymal fates are generated in response to particular growth factors. Substitution of the adipogenic factors insulin and triiodothyronine with bone morphogenetic protein (BMP-4) results in suppression of adipogenesis and development of a mature osteogenic phenotype. In contrast, treatment with transforming growth factor-beta (TGF-beta3) promotes chondrogenic differentiation. Thus, the use of appropriate growth factors and culture milieu steers differentiation of ES cell-derived precursors into distinct mesenchymal compartments. PMID- 15794924 TI - Virus-based reporter systems for monitoring transcriptional activity of hypoxia inducible factor 1. AB - Being key regulator of oxygen homeostasis hypoxia-inducible factor 1 (HIF-1) plays significant roles in cancer progression as well as in cardiovascular diseases. The modulation of HIF-1alpha activity in vivo may represent a valuable therapeutic approach to these disorders. In order to monitor HIF-1 transcriptional activity, we have developed HIF-1alpha-responsive reporter constructs, in which lacZ gene expression is driven by minimal Hsp70 gene promoter or minimal immediate early promoter of cytomegalovirus (CMV) and a combination of hypoxia response elements from regulatory regions of PGK1, ENO1 and LDHA genes. For the efficient delivery to a wide variety of cell types we chose retroviral and lentiviral vectors as carriers of the reporter cassette. We demonstrate that the obtained reporter system i) has a high inducibility in response to treatments leading to HIF-1alpha activation, ii) shows upregulation in response to HIF-1 activation and downregulation following inhibition of HIF 1alpha expression by small interfering RNA, iii) follows the dynamics of endogenous HIF-1 target gene expression. The retrovirus- and lentivirus-based reporters can be used for high-throughput screening of HIF-1alpha modulators and for the study of crosstalk between HIF-1 and different related signal transduction pathways. Potential applications for the reporters are discussed. PMID- 15794926 TI - Bone histomorphometry in 22 male patients with normocalciuric idiopathic osteoporosis. AB - Osteoporosis is associated with increased bone resorption together with a decrease in bone formation. In women, there is an increase of resorption surfaces and in the number of osteoclasts. These changes, however, are not found in males. This purpose of this study was to examine clinical, laboratory, and histomorphometric data in a series of 22 male patients with primary osteoporosis and normocalciuria undergoing transiliac bone biopsy. All of them gave written informed consent for bone biopsy. Automated biochemical profile, urinary calcium excretion, and bone marker assays were performed. Histomorphometric studies were carried out in transiliac bone biopsies obtained with a Bordier-Lepine needle with previous tetracycline labeling. The histomorphometric values of cancellous bone showed significantly lower values of bone volume and values of osteoid surface (OS/BS) and osteoblast surface (Ob.S/BS), and a modest increase in osteoid thickness (O.Th) without changes in the mineralization lag time or eroded surface in patients compared with controls. In cortical bone, there was a low cortical volume (Ct.V/TV) and cortical width (Ct.Wi) in patients compared with controls, without differences in cortical porosity (Po.V/TV). These results suggest that normocalciuric idiopathic osteoporosis in men is characterized by decreased cancellous osteoblasts and bone turnover. PMID- 15794927 TI - Megakaryocytes modulate osteoblast synthesis of type-l collagen, osteoprotegerin, and RANKL. AB - We have previously reported evidence that megakaryocytes may play a role in bone remodeling, possibly by interactions with cells at the bone surface. To investigate the direct effects of megakaryocytes on osteoblasts, maturing megakaryocytes (CD61 positive cells) were isolated and added to cultures of human osteoblasts. Osteoblasts alone and osteoblasts treated with CD61-negative (non megakaryocytic) cells were used as control cultures. After 48 h in culture, megakaryocytes were removed and osteoblasts immunolocalized for type-1 collagen, osteoprotegerin (OPG), and RANKL expression. Similar cultures were used for RNA extraction with mRNA for Col 1A1, OPG, and RANKL in osteoblasts measured quantitatively by RT-PCR. Osteoblasts cultured alone showed high levels of expression of collagen with 74% (+/-7) of cells staining positively. When cultured with megakaryocytes, the number of positively staining cells remained similar but the intensity of expression was increased 1.54-fold (P < 0.02). OPG was expressed by 32% (+/-6.3) of osteoblasts increasing to 51% (+/-5.5) when cultured in the presence of megakaryocytes (P < 0.01) with a 1.63-fold increase in intensity of expression (P < 0.01). In contrast, osteoblasts cultured with megakaryocytes showed suppression of RANKL expression; 35.6% (+/-5.8) of osteoblasts cultured alone stained positively decreasing to 24.3% (+/-5.3) with a 1.6-fold diminished intensity of expression (P < 0.02). Osteoblasts co-cultured with CD61-negative cells showed no differences in collagen, OPG, or RANKL expression levels compared to osteoblasts cultured alone. mRNA data supported these findings with a 3.1-fold increase in Col 1A1 expression in megakaryocyte treated cultures compared to controls (P < 0.02). Low-level OPG mRNA expression increased 8.14-fold in osteoblasts cultured in the presence of megakaryocytes (P < 0.01), while RANKL expression was suppressed 3.3-fold (P < 0.02). These results demonstrate that in vitro, megakaryocytes have direct effects on osteoblastic production of factors affecting both bone formation and resorption. These data provide further evidence that megakaryocytes may play an important role in bone remodeling. PMID- 15794928 TI - Impact of lowering dialysate calcium concentration on serum bone turnover markers in hemodialysis patients. AB - Loss of renal function perturbs bone metabolism because kidney is a vital organ maintaining homeostasis of calcium and phosphate. In hemodialysis patients, bone diseases are serious complications resulting in fractures and extraosseous calcification. The latest clinical practice guidelines by the National Kidney Foundation (New York, US) recommend a dialysate calcium concentration (D-Ca) of 2.5 mEq/L rather than 3.0 mEq/L to avoid excess calcium load and to prevent subsequent vascular calcification. However, there is no perfect agreement yet about which concentration should be chosen because lowering D-Ca might enhance uncoupled bone resorption. Here, we studied effects of lowering D-Ca from 3.0 to 2.5 mEq/L on bone metabolism in 67 patients. Doses of vitamin D and phosphate binders were kept constant for a 2-month period beginning 1 month before the change in D-Ca, and were adjusted thereafter. In group A [intact parathyroid hormone (iPTH) < 100 pg/ml before the study], serum cross-linked N-terminal telopeptide of type I collagen (NTx) increased immediately after lowering D-Ca and then remained stable. Intact osteocalcin (iOC) increased later along with iPTH, suggesting the improvement of adynamic bone disease which shows a marked decrease in bone turnover without osteoid accumulation. Vitamin D was not dosed up in this group. In group B (100 < or = iPTH < 300), serum NTx increased transiently, which is followed by an increase of iOC but not by a change of iPTH. In group C (300 < or = iPTH), lowering D-Ca allowed us to increase the dose of vitamin D without hypercalcemia, leading to a significant decrease in NTx and iPTH. Overall, serum phosphate increased from 5.4 +/- 1.6 to 6.1 +/- 1.6 mg/dL (P < 0.0001) and serum NTx increased by 1.5-fold (P < 0.0001) 1 month after lowering D-Ca. Over a 3-month period after that, serum phosphate and serum NTx decreased to their basal levels. These indicate that bone resorption predominated over formation for only a short period. In conclusion, a D-Ca of 2.5 mEq/L with adjustment of vitamin D ameliorates metabolic abnormalities of bone which develop under 3.0 mEq/L. PMID- 15794929 TI - Local application of rhTGF-beta2 modulates dynamic gene expression in a rat implant model. AB - Various anabolic agents, including transforming growth factor-beta (TGF-beta), have been shown to enhance intramembranous bone regeneration and strengthen the mechanical connection between implant and host skeleton, a prerequisite for clinical success with orthopedic and dental implants. Mechanisms underlying these observations at the level of the gene have received little attention. A rat model was used to examine levels of gene transcription for 21 "osteogenic" genes by real-time polymerase chain reaction at days 1, 3, 5, 7, 10, 14, and 28 in a control group and a group in which the implant was treated with 1 microg recombinant human TGF-beta2 (n = 42, equally divided among the 2 groups and 7 time points). Genes were chosen to represent three functional categories: (1) growth factors, their receptors and antagonists; (2) bone differentiation markers; and (3) inflammation markers. Examination of the transcription profiles showed that nine genes had up-regulated or down-regulated expression levels without a change in timing and 12 genes had accelerated or delayed expression profiles with or without a concomitant change in maximal or minimal expression. The earliest changes (days 1-3) involved accelerated expression profiles for IGF 1R and VEGF and up-regulation of TGF-beta2, TbetaRI, BMP-2, BMP-7, and Cbfa1. Furthermore, principal components analyses showed that some subsets of genes were co-expressed in both groups, although the temporal relationship of these subsets was altered following growth factor treatment. Thus, in addition to changes in individual transcription profiles, the regulatory connections between sets of co expressed genes may also be affected by exogenously delivered anabolic agents during bone regeneration. PMID- 15794930 TI - Systemic versus local application of gentamicin in prophylaxis of implant-related osteomyelitis in a rat model. AB - Administration of perioperative antibiotic prophylaxis is a routine procedure in orthopedic surgery. Besides systemic prophylaxis, only few techniques are established for local application of antibiotics to reduce infection related to orthopedic implants. The aim of this study was to evaluate the efficacy of locally versus systemically applied gentamicin in a rat model (n = 60). For local application, the antibiotic was delivered from a biodegradable poly(D,L-lactide) (PDLLA) coating of titanium implants. The efficacy of local prophylaxis was compared to a systemic single shot application of gentamicin as well as a combination of both administrations. Half of the animals received a weight adopted single shot application of gentamicin 30 min prior to surgery. At surgery, the medullary cavities of the tibiae were contaminated with Staphylococcus aureus (10(2) colony forming units /CFU) and titanium Kirschner wires were implanted into the medullary canals. The implants were either uncoated, PDLLA coated, or coated with PDLLA + 10% w/w gentamicin. The animals were followed up for 42 days. X-ray examinations were performed; body weight, temperature, and the clinical condition were determined. After sacrifice, infection was evaluated by histological and microbiological analysis. All animals treated with uncoated or PDLLA-coated Kirschner wires without systemic application of the antibiotic developed osteomyelitis and all cultures of implants were tested positive on S. aureus. Implant-related osteomyelitis could be prevented by prophylaxis of systemically applied gentamicin in 15% of animals. In contrast, local application of gentamicin delivered from a PDLLA coating was more effective. Onset of infection could be prevented in 90% of animals treated with gentamicin coated Kirschner wires, and in 80% of the animals that were treated with a combination of local and systemic application. The local application from PDLLA-coated implants might support systemic antibiotic prophylaxis in preventing implant-associated osteomyelitis. PMID- 15794931 TI - Stromal cell-derived factor-1 binding to its chemokine receptor CXCR4 on precursor cells promotes the chemotactic recruitment, development and survival of human osteoclasts. AB - Osteoclasts (Oc) derive from hematopoietic precursors present in the circulation and bone marrow, and they differentiate into multinucleated bone-resorbing cells in response to the dual essential signals receptor activator of NF-kappaB ligand (RANKL) and macrophage-colony stimulating factor (M-CSF) primarily provided by bone marrow stromal cells (BMSC) and osteoblasts (Ob). However, little is known about signals that direct Oc precursors from the circulation into bone or control their migration within the marrow. Stromal cell-derived factor-1 (SDF-1 or CXCL12) is a chemokine highly expressed by bone endothelium, BMSC, and immature Ob that is essential for the normal homing, early development, and survival of various hematopoietic progenitor cells. We investigated whether SDF-1 and its unique chemokine receptor CXCR4 were involved in regulating human Oc precursor chemotaxis, development, function, or survival. CXCR4 was highly expressed by freshly isolated human monocyte (MN) populations, in vitro generated Oc and Oc like cells, and mature Oc isolated from human femoral bones. SDF-1 markedly stimulated the chemotactic recruitment of circulating human MN capable of generating bone-resorptive Oc, leading to a 4-fold increase in Oc formation and greater bone pit resorption after their M-CSF + RANKL induced differentiation compared to spontaneously migrating cells. SDF-1 also directly promoted early (but not later) stages of Oc development via stimulating precursor cell numbers, multinucleated cell fusion, increased cell size, and tartrate-resistant acid phosphatase (TRAP) activity in a similar, but non-additive, fashion to M-CSF + RANKL. While SDF-1 did not cause full development of bone-resorbing Oc or stimulate the resorptive function of mature Oc directly, it also did not interfere with any actions promoted by M-CSF + RANKL. In mature human Oc, SDF-1 proved equally as effective as M-CSF + RANKL for preventing Oc apoptosis induced by cytokine withdrawal. In both cases, Oc survival was accompanied by analogous rises in the mRNA ratios for anti-apoptotic Bcl-xL and Bfl-1 relative to pro apoptotic Bax, and by marked protein suppression of the critical pro-apoptotic signal Bim. These findings demonstrate for the first time that SDF-1 chemoattracts circulating human Oc precursors capable of developing into bone resorptive Oc, and that it can stimulate MN cell fusion and TRAP activity, mimic M-CSF + RANKL in early osteoclastogenic effects, substitute for M-CSF + RANKL in maintaining the survival of mature human Oc, and suppress Oc expression of Bim protein. Thus, high levels of SDF-1 produced by bone endothelium, BMSC, and Ob may selectively target circulating Oc precursors into bone and stimulate their marrow migration into suitable perivascular stromal sites for their early development, RANKL differentiation, and survival. Consequently, SDF-1 may be a key factor linking bone vascular cells, BMSC, Ob, and Oc in the normal homeostatic regulation of bone development and remodeling. PMID- 15794932 TI - Aromatase gene and osteoporosis: relationship of ten polymorphic loci with bone mineral density. AB - Aromatase activity appears to be important for bone homeostasis in postmenopausal women. In fact, therapy with aromatase inhibitors is associated with bone loss and fractures. A common biallelic A/G polymorphism in the 3'-untranslated region (UTR) of CYP19-aromatase gene has been associated with differences in gene transcription and the risk of estrogen-responsive tumors. We explored the relationship of such a polymorphism and other 9 polymorphisms situated within or near CYP19 gene with bone mass. The study group comprised 286 postmenopausal women. DNA was isolated from peripheral blood. Biallelic and insertion/deletion polymorphisms were analyzed with exonuclease assays using TaqMan probes. A microsatellite polymorphism in intron 4 was studied by capillary electrophoresis. Bone mineral density (BMD) was determined by DXA. In this cross-sectional study, the postmenopausal decrease in bone mass appeared to be slower in women with AA genotype in the 3'UTR, than in those with AG or GG genotypes. Consequently, there were significant genotype-related differences in BMD. In women after age of 60, hip T-scores were: AA -1.3 +/- 0.1, AG -1.3 +/- 0.2, GG -1.9 +/- 0.1 (P = 0.002). Lumbar spine T-scores were: AA -1.9 +/- 10.2, AG -2.2 +/- 0.1, GG -3.0 +/- 0.2 (P = 0.001). Moreover, GG genotype showed a trend for lower free estrogen levels. This polymorphism was strongly linked to a tetranucleotide repeat in intron 4, as well as to other biallelic polymorphisms situated between 3'UTR and I.2 promoter regions. They all were associated with BMD. However, biallelic polymorphisms in the extreme 5' region of CYP19 and two polymorphisms in neighbor genes were not associated with BMD. In conclusion, common variations of CYP19-aromatase are associated with differences in BMD that seem to be important from an individual as well as from a population perspective. PMID- 15794933 TI - Target identification and validation in drug discovery: the role of proteomics. AB - Proteomics, the study of cellular protein expression, is an evolving technology platform that has the potential to identify novel proteins involved in key biological processes in the cell that may serve as potential drug targets. While proteomics has considerable theoretical promise, individual cells/tissues have the potential to generate many millions of proteins while the current analytical technologies that involve the use of time-consuming two dimensional gel electrophoresis (2DIGE) and various mass spectrometry (MS) techniques are unable to handle complex biological samples without multiple high-resolution purification steps to reduce their complexity. This can significantly limit the speed of data generation and replication and requires the use of bioinformatic algorithms to reconstitute the parent proteome, a process that does not always result in a reproducible outcome. In addition, membrane bound proteins, e.g., receptors and ion channels, that are the targets of many existing drugs, are not amenable to study due, in part, to limitations in current proteomic techniques and also to these being present in low abundance and thus disproportionally represented in proteome profiles. Subproteomes with reduced complexity have been used to generate data related to specific, hypothesis-driven questions regarding target identification, protein-interaction networks and signaling pathways. However progress to date, with the exception of diagnostic proteomics in the field of cancer, has been exceedingly slow with an inability to put such studies in the context of a larger proteome, limiting the value of the information. Additionally the pathway for target validation (which can be more accurately described at the preclinical level as target confidence building) remains unclear. It is important that the ability to measure and interrogate proteomes matches expectations, avoiding a repetition of the disappointment and subsequent skepticism that accompanied what proved to be unrealistic expectations for the rapid contribution of data based on the genome maps, to biomedical research. PMID- 15794934 TI - Stimulation of CD95-induced apoptosis in T-cells by a subtype specific neutral sphingomyelinase inhibitor. AB - Neutral sphingomyelinase (nSMase) has been supposed to be involved in the activation of anti-apoptotic genes and, thus, could well sustain autoimmune reactions by preventing activation induced death of autoreactive T-cells. When screening cellular extracts for SMase activity in the range between pH 6.5 and 8.5 various murine tissue samples as well as cell lines of murine and human origin displayed peaks of activity, both, at pH 7.0 and 8.0. In contrast, T-cells (human T-cell lymphoma and PHA stimulated murine lymph node cells) and monocytic leukemia cells were lacking SMase activity at pH 8.0. Only one peak of activity was found at pH 7.0. Recently we described an inhibitory compound, C11AG which selectively suppresses nSMase activity. In dose-response assays using cellular extracts the pH 7.0 nSMase turned out to be almost 100-fold more sensitive to the inhibitor than the pH 8.0 nSMase. In Jurkat T-cell lymphoma cells lacking the pH 8.0 nSMase, treatment with C11AG enhanced sensitivity to apoptosis: the concentration of CD95-specific antibody anti-APO1 could be lowered by six-fold in order to induce cell death. Concomitantly the expression of the anti-apoptotic protein A1 was found to be down-regulated. In the joints of arthritic mice, apoptosis of T-cells was stimulated after application of C11AG. Accordingly, C11AG displayed curative effects on experimental arthritis: swelling and inflammation were found to be significantly alleviated. PMID- 15794935 TI - Exploitation of the unusual thermodynamic properties of human myeloperoxidase in inhibitor design. AB - Myeloperoxidase plays a fundamental role in oxidant production by neutrophils. It uses hydrogen peroxide and chloride to catalyze the production of hypochlorous acid (HOCl), which contributes to both bacterial killing and oxidative injury of host tissue. Thus, MPO is an interesting target for anti-inflammatory therapy. Here, based on the extraordinary and MPO-specific redox properties of its intermediates compound I and compound II, we present a rational approach in selection and design of reversible inhibitors of HOCl production mediated by MPO. In detail, indole and tryptamine derivatives were investigated for their ability to reduce compounds I and II and to affect the chlorinating activity of MPO. It is shown that these aromatic one-electron donors bound to the hydrophobic pocket at the distal heme cavity and were oxidized efficiently by compound I (k3), which has a one-electron reduction potential of 1.35 V. By contrast, compound II (E degrees ' of the compound II/ferric couple is 0.97 V) reduction (k4) was extremely slow. As a consequence compound II, which does not participate in the halogenation cycle, accumulated. The extent of chlorinating activity inhibition (IC50) was related to the k3/k4 ratio. The most efficient inhibitors were 5 fluorotryptamine and 5-chlorotryptamine with IC50 of 0.79 microM and 0.73 microM and k3/k4 ratios of 386,000 and 224,000, respectively. The reversible mechanism of inhibition is discussed with respect to the enzymology of MPO and the development of drugs against HOCl-dependent tissue damage. PMID- 15794936 TI - Hydrogen peroxide degradation and selective carbidopa-induced cytotoxicity against human tumor lines. AB - The carcinoid tumor, an uncommon neuroendocrine neoplasm, is associated with serotonin overproduction as is more common small cell lung carcinoma (SCLC). alpha-Methyl-dopahydrazine (carbidopa), an inhibitor of the serotonin synthetic enzyme aromatic-L-amino acid decarboxylase, proved lethal to NCI-H727 lung carcinoid cells as well as NCI-H146 and NCI-H209 SCLC cells, but not to five other human tumor cell lines of differing origins [Gilbert JA, Frederick LM, Ames MM. The aromatic-L-amino acid decarboxylase inhibitor carbidopa is selectively cytotoxic to human pulmonary carcinoid and small cell lung carcinoma cells. Clin. Cancer Res. 2000;6:4365-72]. The mechanism of carbidopa cytotoxicity remained an unanswered question. We present data here that incubation of the catechol carbidopa (100 microM) in RPMI and DMEM culture media yielded molar equivalents of hydrogen peroxide (H2O2) within 2-4 h. Alkaline elution studies revealed carbidopa-dependent single-strand DNA breaks in sensitive carcinoid cells comparable to those induced by similar concentrations of H2O2. Neither compound induced significant DNA damage in carbidopa-resistant NCI-H460 large cell lung carcinoma cells. Furthermore, when carbidopa was incubated with a variety of tumor cell types, not only were decreased media H2O2 concentrations detected in the presence of cells, but cell lines least sensitive to carbidopa degraded exogenous H2O2 more rapidly than did sensitive cells. Implicated in these studies, pyruvate degraded H2O2 in RPMI in a dose- and time-dependent manner and reversed carbidopa-induced cytotoxicity to carcinoid cells. Extracellular pyruvate levels produced per h by resistant large cell lung carcinoma cells averaged four-fold that of sensitive carcinoid cells plated at equal density (24 h time course). Finally, carbidopa exposure (100 microM, 24 h) depleted extracellular pyruvate from sensitive carcinoid cells, but reduced pyruvate levels from resistant NCI-H460 cells less than 17%. PMID- 15794937 TI - Necrotic and apoptotic features of cell death in response to Foscan photosensitization of HT29 monolayer and multicell spheroids. AB - Photodynamic therapy (PDT) is an approved anticancer treatment modality that eliminates unwanted cells by the photochemical generation of reactive oxygen species following absorption of visible light by a photosensitizer, which is selectively taken up by tumor cells. Present study reports the modalities of cell death after photosensitization of human adenocarcinoma HT29 monolayer and spheroid cells with a second generation photosensitizer Foscan. Kinetics of apoptosis and necrosis after Foscan-PDT in monolayer cells determined by flow cytometry using labeling of cleaved poly(ADP-ribose) polymerase (PARP) and staining with propidium iodide (PI) demonstrated that Foscan was not a strong inducer of apoptosis and necrosis was a prevailing mode of cell death. Cytochrome c release (cyt c) and mitochondrial membrane potential (Deltapsim) addressed by flow cytometry technique at different time points post-Foscan-PDT demonstrated that cell photoinactivation was governed by these mitochondrial events. Foscan loaded HT29 multicell spheroids, subjected to irradiation with different fluence rates and equivalent light doses, displayed much better tumoricidal activity at the lowest fluence rate used. Apoptosis, measured by caspase-3 activation was evidenced only in spheroids irradiated with the lowest fluence rate and moderate fluence inducing 65% of cell death. Application of higher fluence rates for the same level of photocytotoxicity did not result in caspase-3 activation. The observation of the fluence rate-dependent modulation of caspase-3 activity in spheroids offers the possibility of regulating the mechanism of direct cell photodamage and could be of great potential in the clinical context. PMID- 15794938 TI - 2-Aminoethoxydiphenyl borate perturbs hormone-sensitive calcium stores and blocks store-operated calcium influx pathways independent of cytoskeletal disruption in human A549 lung cancer cells. AB - Recent studies have identified novel actions for 2-aminoethoxydiphenyl borate (2 APB) in triggering calcium release and enhancing calcium influx induced by the depletion of intracellular calcium stores. In this study, we have examined the effects of 2-APB on the human lung adenocarcinoma A549 cell line, which we have previously shown displays a unique calcium influx response, when ER calcium stores are depleted by thapsigargin (TG) treatment. Here, we show that low concentrations of 2-APB failed to induce the rapid augmentation of TG-activated calcium influx previously reported for other cell types. We observed that store operated calcium (SOC) channels in the A549 cell line exhibited short-term sensitivity to low doses of 2-APB, perhaps reflecting a delayed augmentation of SOC channel activity or the recruitment of 2-APB-insensitive SOC channels. In both intact and permeabilized cells, 2-APB effectively discharged a subset of A549 calcium pools corresponding to the hormone-sensitive intracellular calcium stores. The 2-APB-induced calcium release produced a long-lasting perturbation of the adenosine triphosphate (ATP)-releasable calcium pools, effectively uncoupling ATP-activated calcium release even, when stores are replenished with calcium. In contrast to previous reports, we found that disruption of either the actin or microtubule-based cytoskeleton failed to block the 2-APB-induced effects on calcium signaling in A549 cells. Our study describes novel cytoskeletal independent effects of 2-APB on Ca2+-signaling pathways, revealing differentially sensitive Ca2+-influx pathways and long-term perturbation of hormone-sensitive Ca2+ stores. PMID- 15794939 TI - A spectrophotometric assay for fatty acid amide hydrolase suitable for high throughput screening. AB - Signalling via the endocannabinoids anandamide and 2-arachidonylglycerol appears to be terminated largely through the action of the enzyme fatty acid amide hydrolase (FAAH). In this report, we describe a simple spectrophotometric assay to detect FAAH activity in vitro using the ability of the enzyme to hydrolyze oleamide and measuring the resultant production of ammonia with a NADH/NAD+ coupled enzyme reaction. This dual-enzyme assay was used to determine Km and Vmax values of 104 microM and 5.7 nmol/min/mgprotein, respectively, for rat liver FAAH catalyzed oleamide hydrolysis. Inhibitor potency was determined with the resultant rank order of methyl arachidonyl fluorophosphonate>phenylmethylsulphonyl fluoride>anandamide. This assay system was also adapted for use in microtiter plates and its ability to detect a known inhibitor of FAAH demonstrated, highlighting its potential for use in high throughput screening. PMID- 15794940 TI - Increased Akt protein expression is associated with decreased ceramide content in skeletal muscle of troglitazone-treated mice. AB - Although it is generally believed that thiazolidinediones ameliorate insulin resistance by lowering circulating free fatty acids, direct effects of these drugs in skeletal muscle may also contribute to their antidiabetic action. We report that troglitazone administration to mice for 1 day increased the protein expression of Akt (two-fold induction, P<0.001) in skeletal muscle without significant changes in the levels of free fatty acids in plasma. Increased Akt protein expression was associated with reduced phospho-AMP-activated protein kinase abundance and with a fall in the phosphorylation of acetyl-CoA carboxylase, which in turn resulted in an increase in the content of muscular malonyl-CoA (2.4-fold, P<0.05) and lactate (1.4-fold, P<0.05). Troglitazone treatment did not affect the mRNA levels of either Akt1 or Akt2, suggesting that a transcriptional mechanism was not involved, but caused a dramatic reduction in the content of muscular ceramides (76%, P<0.001), lipid-derived second messengers known to increase Akt degradation. Our data indicate that troglitazone treatment inhibited de novo ceramide synthesis, since the content of its precursor, palmitoyl-CoA, was reduced (55%, P=0.05). These results were confirmed in C2C12 myotubes, where troglitazone treatment increased Akt protein expression and prevented the reduction of this protein and the increase in ceramide levels caused by palmitate. These findings implicate ceramide as an important intermediate in the regulation of Akt after troglitazone treatment. PMID- 15794941 TI - Curcumin-induced histone hypoacetylation: the role of reactive oxygen species. AB - Curcumin (Cur), a well-known dietary pigment derived from Curcuma longa, is a promising anticancer drug, but its in vivo target molecules remain to be clarified. Here we report that exposure of human hepatoma cells to Cur led to a significant decrease of histone acetylation. Histone acetyltransferase (HAT) and histone deacetylase (HDAC) are the enzymes controlling the state of histone acetylation in vivo. Cur treatment resulted in a comparable inhibition of histone acetylation in the absence or presence of trichostatin A (the specific HDAC inhibitor), and showed no effect on the in vitro activity of HDAC. In contrast, the domain negative of p300 (a most potent HAT protein) could block the inhibition of Cur on histone acetylation; and the Cur treatment significantly inhibited the HAT activity both in vivo and in vitro. Thus, it is HAT, but not HDAC that is involved in Cur-induced histone hypoacetylation. At the same time, exposure of cells to low or high concentrations of Cur diminished or enhanced the ROS generation, respectively. And the promotion of ROS was obviously involved in Cur-induced histone hypoacetylation, since Cur-caused histone acetylation and HAT activity decrease could be markedly diminished by the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) or their combination, but not by their heat-inactivated forms. The data presented here prove that HAT is one of the in vivo target molecules of Cur; through inhibiting its activity, Cur induces histone hypoacetylation in vivo, where the ROS generation plays an important role. Considering the critical roles of histone acetylation in eukaryotic gene transcription and the involvement of histone hypoacetylation in the lose of cell viability caused by high concentrations of Cur, these results open a new door for us to further understand the molecular mechanism involved in the in vivo function of Cur. PMID- 15794942 TI - Involvement of uracil nucleotides in protection of cardiomyocytes from hypoxic stress. AB - Cardiomyocytes express one or more subtypes of P2 receptors for extracellular nucleotides. P2 purinoceptors, which are activated by nucleotides, are classified as P2X or P2Y: P2X receptors are ligand-gated intrinsic ion channels, and P2Y receptors are G protein-coupled receptors. Extracellular pyrimidine and purine nucleotides are released from the heart during hypoxia. Although the cardioprotective effects of purines acting via purinoceptors were studied intensively, the physiological role of uracil nucleotide-responsive P2Y2, P2Y4, P2Y6, and P2Y14 receptors is still unclear, especially in the cardiovascular system. This study revealed that uridine-5'-triphosphate (UTP) protected cultured rat cardiomyocytes during hypoxia and explored the UTP signaling pathway leading to this cardioprotection. We found that UTP, but not UDP or uridine, significantly reduced cardiomyocyte death induced by hypoxia. Incubation with UTP for 1 h, before exposure to hypoxic conditions, protected the cells 24 h later. The cardioprotective effect of UTP was reduced in the presence of the P2 antagonist suramin. In addition, UTP caused a transient increase of [Ca2+]i in cardiomyocytes. Pyridoxal-5'-phosphate-6-azophenyl-2,4-disulfonate (PPADS) or Reactive blue 2 (RB-2), other antagonists of P2 receptors, abolished the [Ca2+]i elevation caused by UTP. We used various inhibitors of the Ca2+ signaling pathway to show that UTP elevated levels of [Ca2+]i, originating from intracellular sources, via activation of phospholipase C and the IP3 receptor. Interestingly, these inhibitors of the Ca2+ signaling pathway did not prevent the immediate protective effect caused by UTP. Although mitochondrial KATP channels are involved in other preconditioning mediator pathways, the involvement of these channels in the cardioprotective effect induced by UTP was ruled out, because 5 hydroxydecanoic acid (5-HD), a specific inhibitor of these channels, did not prevent the protection. PMID- 15794943 TI - Stimulation of intracellular Ca2+ elevation in neutrophils by thiol-oxidizing phenylarsine oxide. AB - Phenylarsine oxide (PAO), a trivalent arsenical compound, stimulated [Ca2+]i elevation in rat neutrophils in a Ca2+-containing medium but caused no appreciable response in a Ca2+-free medium. PAO also induced external Mn2+ entry, which was inhibited by N-acetyl-L-cysteine (NAC), but failed to elicit any appreciable Ba2+ and Sr2+ entry. Pretreatment of neutrophils with thiol-reducing agents including dithiothreitol (DTT), NAC, 2,3-dimercapto-1-propanol (DMP), 2,3 dimercaptopropane-1-sulfonic acid (DMPS) and tris-(2-carboxyethyl)phosphine (TCEP), all greatly inhibited PAO-induced [Ca2+]i elevation. Addition of Ni2+ or La3+ followed by PAO stimulation also attenuated the Ca2+ signals in a concentration-dependent manner. PAO had no significant effect on the production of reactive oxygen intermediates (ROI) and nitric oxide (NO) nor did it decrease cellular low molecular weight thiols levels. PAO-induced [Ca2+]i elevation was significantly inhibited by 1-[6-[17beta-3-methoxyestra-1,3,5(10)-trien-17 yl]amino]hexyl]-1H-pyrrole-2,5-dione (U-73122), the inhibitor of phospholipase C coupled processes, genistein, a general tyrosine kinase inhibitor, phorbol 12 myristate 13-acetate (PMA), a protein kinase C (PKC) activator, calyculin A, a cortical actin stabilizer, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY 294002), a phosphoinositide 3-kinase inhibitor, 1-[beta-[3-(4 methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole (SKF-96365), and cis-N-(2 phenylcyclopentyl)azacyclotridec-1-en-2-amine (MDL-12,330A), the blockers of receptor-gated and store-operated Ca2+ channels, whereas there was no appreciable effect exerted by aristolochic acid, a phospholipase A2 inhibitor, 7 nitroindazole and N-(3-aminomethyl)benzylacetamidine (1400W), the blockers of NO synthase, and by suspension in a Na+-deprived medium. In contrast, 2 aminoethoxydiphenyl borane (2-APB), the blocker of IP3 receptor and Ca2+ influx, enhanced the PAO-induced response. PAO had no effect on the plasma membrane Ca2+ ATPase (PMCA) activity in the pharmacological isolated neutrophil preparation and the neutrophil membrane fractions. These results indicate that PAO stimulates [Ca2+]i rise in rat neutrophils mainly through the oxidation of vicinal thiol groups on the cell surface membrane to activation of a non-store operated Ca2+ entry (non-SOCE) without affecting the activity of PMCA and the plasmalemmal Na+/Ca2+ exchanger. PMID- 15794944 TI - Cys2,7EtalphaCGRP is a potent agonist for CGRP1 receptors in SK-N-MC cells. AB - The present study reveals that cystein2,7 ethyl-amidealphaCGRP (Cys2,7EtalphaCGRP), an advertised calcitonin gene-related peptide 2 (CGRP2) receptor subtype-selective agonist, is also a potent agonist for the calcitonin gene-related peptide 1 (CGRP1) receptors natively expressed in the SK-N-MC human neuroblastoma cell line. Cys2,7EtalphaCGRP and alpha calcitonin gene-related peptide (alphaCGRP) promote cyclic AMP accumulation in intact SK-N-MC cells to the same extent with EC50 of 1.6+/-0.2 and 0.4+/-0.08 nM, respectively. The antagonist alpha calcitonin gene-related peptide-8-37 (alphaCGRP-(8-37)) produces a concentration-dependent rightward shift of the alphaCGRP- and Cys2,7EtalphaCGRP concentration-response curves with KB-values (71+/-33 and 47+/-21 nM, respectively). The competitive antagonism by alphaCGRP-(8-37) and the similar KB values suggests that alphaCGRP and Cys2,7EtalphaCGRP stimulate the same receptor. In competition binding studies with [125I]-alphaCGRP on SK-N-MC cell membranes, Cys2,7EtalphaCGRP and alphaCGRP-(8-37) display high affinity for the majority of the binding sites with Ki-values of 0.030+/-0.013 and 0.60+/-0.013 nM, respectively. The present findings are at odds with the proclaimed utilization of Cys2,7EtalphaCGRP as a CGRP2 receptor-selective pharmacological tool. Differences between the agonistic profile of this ligand in this and other experimental systems might be species--or even cell type--dependent. PMID- 15794945 TI - Cyclooxygenation of the arachidonoyl side chain of 1-arachidonoylglycerol and related compounds block their ability to prevent anandamide and 2-oleoylglycerol metabolism by rat brain in vitro. AB - In the present study, the abilities of cyclooxygenated derivatives of 1 arachidonoylglycerol and related compounds to prevent the metabolism of [3H]2 oleoylglycerol and [3H]anandamide by cytosolic and membrane fractions, respectively, have been investigated. For each compound, nine concentrations (range 0.2-100 microM) were tested. 1-Arachidonoylglycerol inhibited the hydrolysis of [3H]2-oleoylglycerol with a pI50 value of 5.17+/-0.04 (maximum attainable inhibition 88%). In contrast, the 1-glyceryl esters of prostaglandin D2, E2 and F2alpha were very weak inhibitors of this hydrolysis. Similarly, prostaglandin D2, prostaglandin D2 ethanolamide and prostaglandin D2 serinol amide produced <20% inhibition of [3H]2-oleoylglycerol metabolism at any concentration tested, in contrast to previous data with arachidonic acid, anandamide and arachidonoyl serinol which are all able to inhibit metabolism of this substrate under the assay conditions used here. A similar pattern was seen for all the compounds with respect to the inhibition of [3H]anandamide hydrolysis by the membrane fractions. Thus, cyclooxygenation of the arachidonoyl side chain greatly reduces the ability of 1-arachidonoylglycerol and related compounds to inhibit the hydrolysis of [3H]2-oleoylglycerol and [3H]anandamide. PMID- 15794946 TI - Galphaq potentiation of adenylate cyclase type 9 activity through a Ca2+/calmodulin-dependent pathway. AB - Adenylate cyclase (EC 4.6.1.1) type 9 (AC9) activity has been shown to be inhibited by PMA activation of novel protein kinase C (nPKC) isoforms. In the current study the effect on AC9 activity of activating PKC in physiological relevant manner was examined. Contrary to the anticipated inhibitory effect of activating PKCs through Gq-coupled receptors, activation of transiently expressed Gq-coupled serotonin 5-HT2A or muscarinic M5 receptors resulted in the potentiation of isoproterenol-stimulated cyclic AMP accumulation in HEK293 cells stably expressing AC9 (HEK-AC9). Consistent with Gq-mediated activation of PKC, the addition of the PKC inhibitor bisindolylmaleimide further potentiated isoproterenol-stimulated cyclic AMP accumulation. Expression of a constitutively active mutant of Galphaq in HEK-AC9 cells also produced an enhancement in basal and isoproterenol-stimulated cyclic AMP accumulation. We also examined the role of Galphaq-mediated release of intracellular Ca2+ on the observed potentiation of AC9 activity, by depleting intracellular Ca2+ stores with thapsigargin. In Ca2+ depleted HEK-AC9 cells, activation of transiently expressed M5 receptors resulted in inhibition of isoproterenol-stimulated cyclic AMP accumulation that was blocked by bisindolylmaleimide, indicating that M5 potentiation of AC9 activity requires Ca2+. This prompted us to examine the effects of the calmodulin antagonist W7 and the Ca2+/calmodulin-dependent kinase II (CaMK II) inhibitor KN 93. Pretreating cells with W7 and KN-93 significantly inhibited M5-mediated potentiation of isoproterenol-stimulated cyclic AMP accumulation in HEK-AC9 cells, suggesting that Galphaq potentiation of AC9 activity involves Ca2+/calmodulin and CaMK II. This data provides evidence for Ca2+-mediated potentiation of AC9 activity. PMID- 15794947 TI - cGMP (guanosine 3',5'-cyclic monophosphate) transport across human erythrocyte membranes. AB - Human erythrocytes produce cGMP that can be eliminated by phosphodiesterases or active efflux transporters. The efflux can be studied under controlled conditions as ATP-dependent uptake into inside-out membrane vesicles. However, widely differing values for the transport rates have been reported. We have here examined factors that influence the uptake rates measured and thus may explain these discrepancies. Both the ionic composition of the buffer used during uptake and the mode of vesicle preparation were found to affect the observed transport rates. Furthermore it was apparent that different blood donors expressed on their erythrocytes different amounts of both MRP4 and MRP5, transporters that have been putatively linked to cGMP efflux across erythrocyte membranes. These differences in expression were reflected in differences in rates of cGMP uptake into inside out erythrocyte membrane vesicles. Calculations based on the transport rates observed using vesicles suggest that efflux may be the principal means for eliminating cGMP from human erythrocytes. PMID- 15794948 TI - Induction of apoptosis and modulation of production of reactive oxygen species in human endothelial cells by diphenyleneiodonium. AB - Diphenyleneiodonium (DPI) inhibits activity of flavoenzymes like NADPH oxidase, the major source of superoxide anion in cardiovascular system, but affects also other oxidoreductases. Contradictory data have been published concerning the effect of diphenyleneiodonium on the production of reactive oxygen species in cells, both inhibitory and stimulatory action of DPI being reported. We have examined the effect of DPI on the cellular production of reactive oxygen and nitrogen species (ROS/RNS) and on the proliferation and apoptosis of human vascular endothelial cells. We found increased oxidation of ROS-sensitive probes (dihydrorhodamine 123 and 2',7'-dichlorodihydrofluorescein diacetate) when DPI (20 microM-100 microM) was present in the treated cells. However, oxidation of the fluorogenic probes was inhibited if DPI (20 microM-100 microM) was removed from the reaction medium after cell preincubation. These results suggest an artifactual oxidation of the fluorogenic probes by DPI or its metabolites. A similar pattern of influence of DPI on the production of NO (measured with 4 amino-5-methylamino-2',7'-difluorofluorescein diacetate) was observed. Modulation of generation of reactive oxygen and nitrogen species in DPI-treated cells influenced the nitration of tyrosine residues of cellular proteins, estimated by Western blotting. Decreased level of nitration generally paralleled the lowered production of ROS. A decreased 3-(4,5-dimethylthiazolyl)-3-3(4-sulphophenyl) tetrazolium (MTT) reducing activity of cells for was observed immediately after 1h treatment of human endothelial cells with DPI (1 microM-100 microM), in spite of lack of changes in cell viability estimated by other methods. These results point to a next limitation of MTT in estimation of viability of cells treated with oxidoreductase inhibitors. DPI inhibited the proliferation of HUVECs as well as immortalized cell line HUVEC-ST, as assessed by acid phosphatase activity test and measurement of total nucleic acid content. Proapoptotic action of DPI was observed 12 h after incubation with this compound. PMID- 15794949 TI - A downstream role for protein kinase Calpha in the cytosolic phospholipase A2 dependent protective signalling mediated by peroxynitrite in U937 cells. AB - Exposure to an otherwise non-toxic concentration of peroxynitrite (ONOO-) promotes toxicity in U937 cells supplemented with pharmacological inhibitors of protein kinase C (PKC). This effect is not associated with enhanced formation of H2O2 and is in fact causally linked to inhibition of the cytoprotective signalling driven by endogenous arachidonic acid (AA). The outcome of various approaches using PKC or phospholipase A2 inhibitors, cytosolic phospholipase A2 or PKCalpha antisense-oligonucleotide-transfected cells and supplementation with exogenous AA or tetradecanoylphorbol acetate, as well as PKC down-regulated cells, indicated that ONOO- promotes AA-dependent cytosol to membrane translocation of PKCalpha, an event critical for the cytoprotective signalling under investigation. Evidence for a similar mechanism regulating survival of human monocytes exposed to ONOO- is also presented. These results, along with our previous work on this topic, contribute to the definition of the mechanism whereby monocytes survive to ONOO- in inflamed tissues. PMID- 15794950 TI - Dexamethasone-induced methylprednisolone hemisuccinate hydrolase: its identification as a member of the rat carboxylesterase 2 family and its unique existence in plasma. AB - Carboxylesterases (CESs) play important roles in the metabolism of many ester drugs. In the present study, we identified and characterized dexamethasone induced methylprednisolone hemisuccinate (MPHS) hydrolase in rat liver microsomes. Intraperitoneal injection of dexamethasone resulted in a significant increase in the level of MPHS hydrolase activity accompanied by induction of a specific CES isozyme. Since the biochemical characteristics of the induced CES isozyme were very similar to those of rat CES RL4, we hypothesized that these were the same enzymes. The results of nano-electrospray ionization tandem mass spectrometry analysis revealed that both dexamethasone-induced CES isozyme and CES RL4 possessed identical peptide fragments to those of , a rat CES2 isozyme, supporting our hypothesis. Furthermore, the results of reverse transcription polymerase chain reaction showed that the amount of mRNA in dexamethasone-treated liver was greater than that in control liver. To confirm that encodes dexamethasone-induced CES isozyme, cDNA cloning was performed and the obtained cDNA was expressed in Sf9 cells by using a baculovirus-mediated expression system. The recombinant CES protein could hydrolyze MPHS and exhibited biochemical characteristics similar to those of CES RL4. Collectively, the results indicated that dexamethasone-induced MPHS hydrolase in liver microsomes is a rat CES2 isozyme. Interestingly, the results also showed that this rat CES2 isozyme exists in plasma and that the amount of this protein is increased by dexamethasone. These findings, together with the findings described above, provide important information for the study of phramacokinetics and pharmacodynamics of ester-drugs as well as for the study of CESs. PMID- 15794951 TI - Vaccine trial as "probe" to define the burden of pneumococcal pneumonia disease. PMID- 15794952 TI - Making the world's children count. PMID- 15794953 TI - Paediatric femoral fracture. PMID- 15794954 TI - US exceptionalism comes to research ethics. PMID- 15794955 TI - Listening to consumers and HIV vaccine preparedness. PMID- 15794956 TI - Commission for Africa: can it make a difference to health? PMID- 15794958 TI - Mantombazana Tshabalala-Msimang: South Africa's controversial health minister. PMID- 15794960 TI - Mortality in Iraq. PMID- 15794961 TI - Does medicine have a moral message? PMID- 15794962 TI - Mortality in Iraq. PMID- 15794963 TI - International aid, partnership, and child survival. PMID- 15794964 TI - A new leadership for UNICEF: do values matter? PMID- 15794965 TI - Repeat sudden unexpected infant deaths. PMID- 15794966 TI - Repeat sudden unexpected infant deaths. PMID- 15794968 TI - Efficacy of nine-valent pneumococcal conjugate vaccine against pneumonia and invasive pneumococcal disease in The Gambia: randomised, double-blind, placebo controlled trial. AB - BACKGROUND: Pneumonia is estimated to cause 2 million deaths every year in children. Streptococcus pneumoniae is the most important cause of severe pneumonia. We aimed to assess the efficacy of a nine-valent pneumococcal conjugate vaccine in children. METHODS: We undertook a randomised, placebo controlled, double-blind trial in eastern Gambia. Children age 6-51 weeks were randomly allocated three doses of either pneumococcal conjugate vaccine (n=8718) or placebo (8719), with intervals of at least 25 days between doses. Our primary outcome was first episode of radiological pneumonia. Secondary endpoints were clinical or severe clinical pneumonia, invasive pneumococcal disease, and all cause admissions. Analyses were per protocol and intention to treat. FINDINGS: 529 children assigned vaccine and 568 allocated placebo were not included in the per-protocol analysis. Results of per-protocol and intention-to-treat analyses were similar. By per-protocol analysis, 333 of 8189 children given vaccine had an episode of radiological pneumonia compared with 513 of 8151 who received placebo. Pneumococcal vaccine efficacy was 37% (95% CI 27-45) against first episode of radiological pneumonia. First episodes of clinical pneumonia were reduced overall by 7% (95% CI 1-12). Efficacy of the conjugate vaccine was 77% (51-90) against invasive pneumococcal disease caused by vaccine serotypes, 50% (21-69) against disease caused by all serotypes, and 15% (7-21) against all-cause admissions. We also found an efficacy of 16% (3-28) against mortality. 110 serious adverse events arose in children given the pneumococcal vaccine compared with 131 in those who received placebo. INTERPRETATION: In this rural African setting, pneumococcal conjugate vaccine has high efficacy against radiological pneumonia and invasive pneumococcal disease, and can substantially reduce admissions and improve child survival. Pneumococcal conjugate vaccines should be made available to African infants. PMID- 15794969 TI - WHO estimates of the causes of death in children. AB - BACKGROUND: Child survival efforts can be effective only if they are based on accurate information about causes of deaths. Here, we report on a 4-year effort by WHO to improve the accuracy of this information. METHODS: WHO established the external Child Health Epidemiology Reference Group (CHERG) in 2001 to develop estimates of the proportion of deaths in children younger than age 5 years attributable to pneumonia, diarrhoea, malaria, measles, and the major causes of death in the first 28 days of life. Various methods, including single-cause and multi-cause proportionate mortality models, were used. The role of undernutrition as an underlying cause of death was estimated in collaboration with CHERG. FINDINGS: In 2000-03, six causes accounted for 73% of the 10.6 million yearly deaths in children younger than age 5 years: pneumonia (19%), diarrhoea (18%), malaria (8%), neonatal pneumonia or sepsis (10%), preterm delivery (10%), and asphyxia at birth (8%). The four communicable disease categories account for more than half (54%) of all child deaths. The greatest communicable disease killers are similar in all WHO regions with the exception of malaria; 94% of global deaths attributable to this disease occur in the Africa region. Undernutrition is an underlying cause of 53% of all deaths in children younger than age 5 years. INTERPRETATION: Achievement of the millennium development goal of reducing child mortality by two-thirds from the 1990 rate will depend on renewed efforts to prevent and control pneumonia, diarrhoea, and undernutrition in all WHO regions, and malaria in the Africa region. In all regions, deaths in the neonatal period, primarily due to preterm delivery, sepsis or pneumonia, and birth asphyxia should also be addressed. These estimates of the causes of child deaths should be used to guide public-health policies and programmes. PMID- 15794970 TI - Treatments for paediatric femoral fractures: a randomised trial. AB - BACKGROUND: Treatments for femoral fractures in children vary widely and have been investigated only in case series. We did a multicentre randomised trial to compare malunion rates after external fixation and after early application of a hip spica cast for paediatric femoral shaft fractures. METHODS: All children aged 4-10 years with femoral fractures, admitted to four paediatric hospitals, were randomly assigned early application of hip spica or external fixation. The primary outcome was malunion at 2 years after the fracture. Secondary outcomes were scores on the RAND physical function child health questionnaire and the post hospitalisation behavioural questionnaire, and parents' and children's ratings of overall satisfaction with treatment. Analysis was by intention to treat based on children who reached the 2 year evaluation. FINDINGS: Of 60 children assigned to the hip-spica group, 56 reached the 2-year assessment; of them, six (11%) required other forms of treatment because of unacceptable loss of reduction. Of 48 children assigned external fixation, 45 reached the 2-year assessment; two (4%) had refractures and five (11%) required operative adjustment of the fixator. The rate of malunion was significantly higher in the hip-spica group than in the external-fixator group (25/56 [45%] vs 7/45 [16%]; 95% CI for difference 12-46%; p=0.002). The two groups had similar mean scores for the RAND physical function health questionnaire (0.34 vs 0.45; 95% CI for difference, -0.57 to 0.34; p=0.61), for the post-hospitalisation questionnaire (106.8 vs 106.3; -4.9 to 5.9; p=0.86), and for parents' satisfaction (4.3 vs 4.2; -0.3 to 0.6; p=0.5) and children's ratings of happiness with treatment (6.9 vs 7.7; -2.2 to 0.5; p=0.21). INTERPRETATION: Early application of hip spica has a small role in the treatment of paediatric femoral fractures. Future trials need to compare external fixation with flexible intramedullary nails. PMID- 15794971 TI - Epidemiology and reporting of randomised trials published in PubMed journals. AB - Although randomised trials are important for evidence-based medicine, little is known about their overall characteristics. We assessed the epidemiology and reporting of methodological details for all 519 PubMed-indexed randomised trials published in December, 2000 (383 [74%] parallel-group, 116 [22%] crossover). 482 (93%) were published in specialty journals. A median of 80 participants (10th 90th percentile 25-369) were recruited for parallel-group trials. 309 (60%) were blinded. Power calculation, primary outcomes, random sequence generation, allocation concealment, and handling of attrition were each adequately described in less than half of publications. The small sample sizes are worrying, and poor reporting of methodological characteristics will prevent reliable quality assessment of many published trials. PMID- 15794972 TI - Deep vein thrombosis. AB - Deep vein thrombosis and its sequelae pulmonary embolism and post-thrombotic syndrome are some of the most common disorders. A thrombus either arises spontaneously or is caused by clinical conditions including surgery, trauma, or prolonged bed rest. In these instances, prophylaxis with low-dose anticoagulation is effective. Diagnosis of deep vein thrombosis relies on imaging techniques such as ultrasonography or venography. Only about 25% of symptomatic patients have a thrombus. Thus, clinical risk assessment and D-dimer measurement are used to rule out deep vein thrombosis. Thrombus progression and embolisation can be prevented by low-molecular-weight heparin followed by vitamin K antagonists. Use of these antagonists for 3-6 months is sufficient for many patients. Those with antithrombin deficiency, the lupus anticoagulant, homozygous or combined defects, or with previous deep vein thrombosis can benefit from indefinite anticoagulation. In cancer patients, low-molecular-weight heparin is more effective than and is at least as safe as vitamin K antagonists. Women seem to have a lower thrombosis risk than men, but pregnancy or use of oral contraceptives or hormone replacement therapy represent important risk factors. PMID- 15794973 TI - Hospital-acquired neonatal infections in developing countries. AB - Hospital-born babies in developing countries are at increased risk of neonatal infections because of poor intrapartum and postnatal infection-control practices. We reviewed data from developing countries on rates of neonatal infections among hospital-born babies, range of pathogens, antimicrobial resistance, and infection control interventions. Reported rates of neonatal infections were 3-20 times higher than those reported for hospital-born babies in industrialised countries. Klebsiella pneumoniae, other gram-negative rods (Escherichia coli, Pseudomonas spp, Acinetobacter spp), and Staphylococcus aureus were the major pathogens among 11,471 bloodstream isolates reported. These infections can often present soon after birth. About 70% would not be covered by an empiric regimen of ampicillin and gentamicin, and many might be untreatable in resource-constrained environments. The associated morbidity, mortality, costs, and adverse effect on future health-seeking behaviour by communities pose barriers to improvement of neonatal outcomes in developing countries. Low-cost, "bundled" interventions using systems quality improvement approaches for improved infection control are possible, but should be supported by evidence in developing country settings. PMID- 15794974 TI - Neonatal survival: a call for action. AB - To achieve the Millennium Development Goal for child survival (MDG-4), neonatal deaths need to be prevented. Previous papers in this series have presented the size of the problem, discussed cost-effective interventions, and outlined a systematic approach to overcoming health-system constraints to scaling up. We address issues related to improving neonatal survival. Countries should not wait to initiate action. Success is possible in low-income countries and without highly developed technology. Effective, low-cost interventions exist, but are not present in programmes. Specific efforts are needed by safe motherhood and child survival programmes. Improved availability of skilled care during childbirth and family/community-based care through postnatal home visits will benefit mothers and their newborn babies. Incorporation of management of neonatal illness into the integrated management of childhood illness initiative (IMCI) will improve child survival. Engagement of the community and promotion of demand for care are crucial. To halve neonatal mortality between 2000 and 2015 should be one of the targets of MDG-4. Development, implementation, and monitoring of national action plans for neonatal survival is a priority. We estimate the running costs of the selected packages at 90% coverage in the 75 countries with the highest mortality rates to be US4.1 billion dollars a year, in addition to current expenditures of 2.0 billion dollars. About 30% of this money would be for interventions that have specific benefit for the newborn child; the remaining 70% will also benefit mothers and older children, and substantially reduce rates of stillbirths. The cost per neonatal death averted is estimated at 2100 dollars (range 1700-3100 dollars). Maternal, neonatal, and child health receive little funding relative to the large numbers of deaths. International donors and leaders of developing countries should be held accountable for meeting their commitments and increasing resources. PMID- 15794975 TI - Trade, drugs, and health-care services. PMID- 15794976 TI - Always trust your gut feeling. PMID- 15794977 TI - Severe anaemia in an 11-month-old girl. PMID- 15794978 TI - Evaluation of an oligonucleotide ligation assay for detection of mutations in HIV 1 subtype C individuals who have high level resistance to nucleoside reverse transcriptase inhibitors and non-nucleoside reverse transcriptase inhibitors. AB - The oligonucleotide ligation assay (OLA) has been proposed as an affordable alternative to sequence-based HIV-1 drug resistance testing in resource poor settings. The aim was to evaluate OLA for detecting mutations K103N, Y181C, K65R, Q151M, M184V and T215Y/F in subtype C. Forty-four subtype C and 8 subtype B HIV-1 positive individuals were analysed using the ViroSeqtrade mark HIV-1 genotyping assay (Applied Biosystems, Foster City, CA). A one-step RT-PCR and nested PCR were performed using subtype B specific primers from the OLA kit (NIH AIDS Research and Reference Reagent Program). Seventy-eight subtype C sequences were used to design subtype C specific primers. Ligation and detection steps were followed according to OLA kit protocol. For codons, K103N, Y181C, K65R, Q151M, M184V and T215Y/F, four or more mismatches compared to the probe or mismatches less than four bases from the ligation site were not tolerated. Results revealed accurate identification of mutations in 2/10, 4/9 3/9, 6/7, 2/7 and 6/7 VQA samples and 5/20, 4/17 0/20, 18/24, 5/24 and 13/24 subtype C positive individuals, respectively. It was concluded that the probes and primers in the NIH reference kit would need modification to optimize detection of mutations in subtype C individuals. PMID- 15794979 TI - Nested multiplex PCR assay for detection of human enteric viruses in shellfish and sewage. AB - Environmental samples and contaminated shellfish present frequently low concentrations of more than one viral species. For this reason, a nested multiplex RT-PCR was developed for the detection of adenoviruses, enteroviruses and hepatitis A viruses in different environmental samples such as urban sewage and shellfish. This assay will save time and cost for detection of these enteric viruses with a smaller sample volume, which otherwise can be a limiting factor in routine analysis. The limit of detection was approximately 1 copy for adenovirus and 10 copies for enterovirus and hepatitis A virus per PCR reaction using titrated cell-cultured viruses as template material. In shellfish and environmental samples, this multiplex PCR was optimized to detect all three viruses simultaneously when the concentration of each virus was equal or lower than 1000 copies per PCR reaction. This is the level found predominantly in the environment and in shellfish when the numbers of fecal bacterial and phage indicators are low. The detection of human adenoviruses by PCR has been suggested as a molecular index of fecal contamination of human origin in the environment and food and the multiplex assay developed may be a tool for evaluating the presence of viral contamination in shellfish and water and to expand microbiological control to include viral markers. PMID- 15794980 TI - Detection of baboon cytomegalovirus (BaCMV) by PCR using primers directed against the glycoprotein B gene. AB - We have cloned and sequenced the glycoprotein B genes from five strains of BaCMV, isolated from three subspecies of cynocephalus baboons (olive, yellow and chacma). Primers were designed using conserved DNA regions of the gB gene to allow DNA amplification from all strains of BaCMV. These regions differ sufficiently from human CMV that HCMV strains are not amplified, thus allowing differentiation of BaCMV from HCMV. These diagnostic primers were used to test crude nucleic acid extracts from 27 strains of BaCMV and detected 26 of them. Overall, the sensitivity and specificity of this assay are 96.7 and 100%, respectively. BaCMV strains isolated from yellow and olive baboons were very similar and could be discriminated from strains isolated from chacma baboons using a second set of PCR primers. Phylogenetic analysis of the gB genes supported the inferred close relationship of strains isolated from olive and yellow baboons. PMID- 15794981 TI - Real Time RT-PCR for the detection and quantitation of bovine respiratory syncytial virus. AB - A quantitative Real Time RT-PCR assay was developed to detect and quantify bovine respiratory syncytial virus (BRSV) in the respiratory tract of infected animals. A pair of primers and a TaqMan probe targeting conserved regions of the nucleoprotein gene of BRSV were designed. The detection limit of the assay was shown to be 10(3) RNA copies and standard curve demonstrated a linear range from 10(3) to 10(8) copies as well as an excellent reproducibility. The efficiency of the BRSV Real Time RT-PCR was then assessed by detecting BRSV in lungs, tracheas and bronchoalveaolar fluids (BAL) samples of experimentally infected calves. The assay was shown to be 100 times more sensitive than conventional RT-PCR and was more efficient for BRSV diagnosis. Finally, the Real Time RT-PCR was used to quantify BRSV load in BAL fluids of four experimentally infected calves for 14 days. The high sensitivity, rapidity and reproducibility of the BRSV Real Time RT PCR make this method suitable for diagnostic and for the evaluation of the efficiency of new vaccines. PMID- 15794982 TI - Specific detection of the PVY(N)-W variant of Potato virus Y. AB - PVY(N)-W is one of the variant populations of Potato virus Y (PVY). This variant is of concern in seed potato production and requires a specific diagnosis since it induces more or less symptomless infections and is not detectable easily in field inspections. Moreover, this variant is serologically indistinguishable from the common strain PVY(O). This study describes a simple and specific molecular detection test for the PVY(N)-W variant using a PCR protocol based on the recombinant point within the HC-Pro/P3 region of PVY(N) variants (PVY(NTN), PVY(N)-W). To avoid both detection of recombinant PVY(NTN) and PVY(N)-W isolates, a forward PVY(N)-like primer located in the HC-Pro region coupled to a reverse PVY(O)-like primer located in the NIa region was designed to amplify a specific PCR product of 4114 nt from PVY(N)-W isolates. This technique was assessed on 41 PVY reference and field isolates. Only isolates referenced as PVY(N)-W were amplified and gave the expected PCR product of 4114 nt, whereas no band was obtained from PVY(N), PVY(NTN) or PVY(O) isolates. In conclusion, this PVY(N)-W diagnosis tool is rapid, easy-to-use and suitable for large-scale testing in laboratories of seed potato certification. PMID- 15794983 TI - A branched, synthetic oligopeptide corresponding to a region of glycoprotein G of HSV-1 reacts sensitively and specifically with HSV-1 antibodies in an ELISA. AB - Herpes simplex viruses types 1 and 2 (HSV-1 and HSV-2), which are common worldwide, are so similar that antibodies directed against one serotype may crossreact with antigens from the other one. Methods for specific detection of antibodies against HSV-1 or HSV-2 are based upon the antigenicities of glycoproteins G. However, due to the cost, the available commercial methods may not readily be used in developing countries. A different enzyme-linked immunosorbent assay (ELISA) method, based upon a synthetic oligopeptide corresponding to an immunogenic region in glycoprotein G of HSV-2, has been used recently and successfully for detection of HSV-2 antibodies. In the present study, the sequences of a newly identified immunogenic and type-specific region in glycoprotein G of HSV-1 was used to synthesize three different, branched oligopeptides. The performances of these peptides in an ELISA were investigated by testing Scandinavian and African sera which were characterized by commercial ELISA and Western blotting methods and divided into four groups either lacking HSV antibodies, containing antibodies against one or the other virus, or against both types. The peptide which corresponded in sequence to the immunodominant region was as specific and sensitive by an ELISA as were the commercial methods. The method is inexpensive and reliable. PMID- 15794984 TI - Phenotypic characterisation of cytomegalovirus DNA polymerase: a method to study cytomegalovirus isolates resistant to foscarnet. AB - A phenotypic method was developed to test mutations in the human cytomegalovirus (HCMV) DNA polymerase gene (UL54) suspected to confer resistance to foscarnet. This method was used to determine the biochemical phenotype of wild-type and mutated HCMV DNA polymerases that had been synthesised in vitro as follows. The UL54 genes were amplified from foscarnet-resistant and -sensitive isolates by PCR and the products were cloned into an expression vector under the control of a T7 promoter. Mutations were introduced by site-directed mutagenesis into wild-type gene UL54 and then polymerases were synthesised by using a commercially available coupled transcription/translation system. Polymerase activity was measured with and without foscarnet by detecting the incorporation of digoxigenin-labelled nucleotides into the growing DNA chain. The results of this non-radioactive assay were consistent with those obtained with the conventional radioactive assay. It was found that the activity of polymerases containing the V715M or E756K mutations was inhibited by foscarnet at concentrations 70- and 30-fold higher than that of wild-type polymerase, respectively. Change N495K and a combination of changes K415R and S291P, both observed in foscarnet-resistant isolates, induced a 5- and 10-fold decrease in susceptibility to foscarnet, respectively. This non-radioactive phenotypic assay could be useful for the characterisation of mutations that confer HCMV resistance to foscarnet. PMID- 15794985 TI - Detection of antibody to the foot-and-mouth disease virus (FMDV) non-structural polyprotein 3ABC in sheep by ELISA. AB - The specificity and sensitivity of an ELISA for detecting IgG to the 3ABC non structural protein of foot-and-mouth disease (FMD) virus was evaluated in FMD naive, aerosol-infected, aerosol plus direct contact infected and field-exposed sheep. All 12 sheep that were experimentally infected without prior vaccination seroconverted in the test, although fewer field sera from FMD-exposed sheep were scored seropositive compared to test results for structural protein antibodies. The 3ABC test specificity was 98 or 100% according to whether sera reacting in the doubtful range were scored as positive or negative. The test was then used to investigate the antibody response of sheep vaccinated against FMD and exposed to the virus by an aerosol challenge 4-14 days later. The response of individual animals varied. Whether immunised with high or low doses of vaccine, the development of 3ABC antibody was most likely in sheep from which live virus was recovered at or beyond 9 days post-challenge. Non-structural responses were also more frequent in animals from which multiple incidences of live FMD virus isolation (perhaps more indicative of true virus replication) were demonstrated. PMID- 15794986 TI - Approaches to Scrapie diagnosis by applying immunohistochemistry and rapid tests on central nervous and lymphoreticular systems. AB - Comparative studies evaluating the performance of Transmissible Spongiform Encephalopathies (TSE) rapid tests (validated for Bovine Spongiform Encephalopathy samples) on Scrapie samples have not been reported widely, particularly those dealing with lymphoreticular system tissues to a much lesser extent. The main objective of this study was to compare the ability of two current rapid tests (Western blot and Luminiscence Immunoassay Prionics-Check; WB and LIA, respectively) to detect PrPsc using central nervous system as well as lymphoreticular system samples corresponding to naturally infected animals. Thirty-four Scrapie-affected sheep, 26 with clinical signs of the disease, were included in the study. Tonsil, retropharyngeal lymph node and medulla oblongata were assessed by three tests: immunohistochemistry (confirmatory test), WB and LIA (rapid tests). The conclusion which can be drawn from this study is the fact that all animals involved in the study, including those at a preclinical stage, could be diagnosed regardless of the test used (with immunohistochemistry consistently showing higher sensitivity) only when the analyses of both the central nervous system and the lymphoreticular system were considered. The choice of these tissues for routine diagnosis is, therefore, proposed as a valuable tool to highly reduce the number of undetected positive cases. PMID- 15794987 TI - Improved serological diagnosis of Poplar mosaic virus with monoclonal antibodies. AB - Poplar mosaic virus (PopMV) is widespread in all countries where poplar is grown, and causes severe economic losses in terms of quantity and quality of wood production. Control is based on indexing, aimed at the production of healthy propagation material. The currently used diagnostic method is double antibody sandwich (DAS) ELISA with polyclonal antibodies, which is relatively simple and inexpensive and more reliable than visual inspection of symptoms in the nurseries. However, this method also has disadvantages, mainly low sensitivity in relation to low concentration and irregular distribution of the virus in the plant. In this study, a new diagnostic method for PopMV based on production and use of a monoclonal antibody (Mab) in a triple antibody sandwich (TAS) ELISA, is presented. The TAS-ELISA with monoclonal antibodies was optimised by testing a range of reagent combinations and concentrations. PopMV was detected by the optimised TAS-ELISA with sensitivity more than 100 times higher than by DAS-ELISA with polyclonal antibodies. Six PopMV isolates from four European countries were detected with the same efficiency, indicating that no limitations to the practical use of the TAS-ELISA arise due to excessive epitope-specificity of the monoclonal antibody employed. PMID- 15794989 TI - An improved Western blot assay to assess the clearance of prion protein from plasma-derived therapeutic proteins. AB - Specific detection of the pathogenic prion protein, PrP(Sc), is essential for determining the prion clearance capacity of purification processes for therapeutic proteins. Use of a previously described indirect (two-antibody) Western blot assay sometimes resulted in the appearance of non-specific protein bands that interfered with the detection of small amounts of PrP(Sc)-specific signal, limiting the amount of clearance that could be determined for steps so affected. It is shown that these non-specific signals are due to the interaction between immunoglobulin fragments in the sample and the secondary antibody used in the assay. To circumvent this problem, a direct Western blot assay using a prion specific primary antibody conjugated to the reporter enzyme alkaline phosphatase was developed. Application of the direct Western blot assay resulted in a significant reduction of non-specific signal while retaining the detection sensitivity for PrP(Sc)-specific signal. Therefore, the direct Western blot assay format is an improved tool for determining prion clearance capacity, particularly for immunoglobulin-rich samples. PMID- 15794988 TI - Recombinant nucleocapsid protein-based IgG enzyme-linked immunosorbent assay for the serological diagnosis of SARS. AB - The recombinant nucleocapsid protein (rNP) of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) was expressed in a baculovirus system. The purified SARS-CoV rNP was used as an antigen for detection of SARS-CoV antibodies in IgG enzyme-linked immunosorbent assay (ELISA). The ELISA was evaluated in comparison with neutralizing antibody assay and the authentic SARS-CoV antigen-based IgG ELISA. Two-hundred and seventy-six serum samples were collected from health care workers in a hospital in which a nosocomial SARS outbreak took place and used for evaluation. The SARS-CoV rNP-based IgG ELISA has 92% of sensitivity and specificity compared with the neutralizing antibody assay and 94% sensitivity and specificity compared with the authentic SARS-CoV antigen-based IgG ELISA. The results suggest that the newly developed SARS-CoV rNP-based IgG ELISA is a valuable tool for the diagnosis and seroepidemiological study of SARS. The SARS CoV rNP-based IgG ELISA has an advantage over the conventional IgG ELISA in that the antigen can be prepared by laboratory workers without the risk of infection. PMID- 15794990 TI - Sequence analysis of murine leukemia virus envelope gene from inoculated mice. AB - Introduction of amino acids substitutions in murine leukemia virus genome is a powerful method to determine the relative importance of various viral factors in pathogenesis. However, introduction of such amino acids substitution could result in viruses at a selective disadvantage, and eventual selection of revertants. It is thus essential to verify if the mutation is maintained stably in replicating virus and in infected tumor cells. In the present study, viral nucleic acid sequences from diseased animals were determined using different approaches. Small blood samples were found adequate for direct RNA extraction and reverse transcriptase-PCR amplification followed by automated DNA sequencing. Alternatively, replication-competent viruses were recovered specifically by applying blood samples onto permissive cells; viral RNA is then extracted from tissue culture medium and similarly sequenced. Tissue samples were also used to amplify viral sequences from tumors DNA while small pieces of tumors tissues were applied onto permissive cells to isolate replicating viruses. The combined experimental approach was used to show sequence conservation using a mutant altered in the intracytoplasmic region of viral envelope glycoprotein. No difference was observed between viruses recovered directly from the animal and those amplified onto cultured cells. PMID- 15794991 TI - Human apoE targeted replacement mouse lines: h-apoE4 and h-apoE3 mice differ on spatial memory performance and avoidance behavior. AB - Apolipoprotein E4 (apoE4), one of the three most common human apoE (h-apoE) isoforms, is a major genetic risk factor for Alzheimer's disease and for cognitive deficits associated with aging. The biological mechanisms involving apoE in learning and memory processes are unclear. A potential isoform-dependent effect of h-apoE on cognitive performance was studied in gene-targeted mice, which show physiological expression levels and distribution of h-apoE3 or h apoE4. Male and female h-apoE3 and h-apoE4, apoE-deficient and C57BL/6J mice (4-5 months) were subjected to tasks evaluating spatial memory and avoidance conditioning. Female h-apoE4 mice did not detect changes in the spatial configuration of objects as opposed to female h-apoE3 mice. Female h-apoE3 mice failed to improve their performance during training in a reference memory version of the spatial water-maze task, but performed well during the probe trial 24 h after the last training trial. Memory retention performances of h-apoE4 mice were impaired during this probe trial. Both h-apoE3 and h-apoE4 mice did not improve their performance in a water-maze delayed matching to place task. Finally, h apoE3 mice showed mild perturbations in a Y-maze active avoidance task, whereas both h-apoE mouse lines performed well in a passive avoidance task. Thus, spatial memory performances appeared particularly sensitive to h-apoE-isoform-dependent effects. Deficits occurred predominantly in female h-apoE4 mice, which support the hypothesis that humans carrying h-apoE4, especially women, have impaired spatial memory compared to those carrying h-apoE3. PMID- 15794992 TI - Impaired memory and olfactory performance in NaSi-1 sulphate transporter deficient mice. AB - In the present study, NaSi-1 sulphate transporter knock-out (Nas1-/-) mice, an animal model of hyposulphataemia, were examined for spatial memory and learning in a Morris water maze, and for olfactory function in a cookie test. The Nas1-/- mice displayed significantly (P<0.05) increased latencies to find an escape platform in the reversal learning trials at 2 days but not 1 day after the last acquisition trial in a Morris water maze test, suggesting that Nas1-/- mice may have proactive memory interference. While the wild-type (Nas1+/+) mice showed a significant (P<0.02) decrease in time to locate a hidden food reward over four trials after overnight fasting, Nas1-/- mice did not change their performance, resulting in significantly (P<0.05) higher latencies when compared to their Nas1+/+ littermates. There were no significant differences between Nas1-/- and Nas1+/+ mice in the cookie test after moderate food deprivation. In addition, both Nas1-/- and Nas1+/+ mice displayed similar escape latencies in the acquisition phase of the Morris water maze test, suggesting that learning, motivation, vision and motor skills required for the task may not be affected in Nas1-/- mice. This is the first study to demonstrate an impairment in memory and olfactory performance in the hyposulphataemic Nas1-/- mouse. PMID- 15794994 TI - 'Looping'-an exploration mechanism in a dark open field. AB - The behavior of Tristram's jird (a species of gerbil) in an illuminated open field resembled that of other rodents, comprising round trips to a home base and alternating between periods of progression (locomoting) and of stopping. In this study, we compared the characteristics of exploration in a dark arena with exploration by the same individuals in a lit arena. In the dark arena, stopping episodes were brief and fewer, suggesting almost continuous locomotion by the rodents. The clear distinction between progression and stopping that had characterized locomotion in an illuminated arena, thus diminished in the dark. There was also no apparent home base in the dark and traveling consisted in moving in a circular path, closing a loop to a recently traveled place that varied from one loop to the next. Locomotion in the dark may thus be regarded as a set of loops (round trips) to a continuously shifting home base, whereas with lights on the round trips converge to a home base using visible environmental landmarks. We suggest that a similar looping mechanism may be applicable to the behavior of hippocampal rats displaying hyperactivity and diversified locomotion, reminiscent of that seen in jirds in a dark arena. PMID- 15794993 TI - Exercise, but not environmental enrichment, improves learning after kainic acid induced hippocampal neurodegeneration in association with an increase in brain derived neurotrophic factor. AB - Previous studies have suggested that exercise in a running wheel can be neuroprotective, perhaps due to, among others, gene-expression changes after exercise, increases in trophic proteins and/or enhanced cardiovascular responsivity. Here we ask whether physical exercise or environmental enrichment provide protection after brain damage, especially in terms of recovery of cognitive function. To evaluate the neuroprotective effect of these conditions, we used the kainic acid (KA) model of neuronal injury. Systemically-administered KA induces excitotoxicity by overstimulation of glutamate receptors, resulting in neuronal death by necrosis and apoptosis. Our results show that exercise, but not enriched environment, prior to KA-induced brain damage, improved behavioural performance in both Morris watermaze and object exploration tasks. However, prior exercise did not decrease to control levels the hyperactivity normally seen in KA treated animals, as measured by ambulation in the open field. Furthermore, both exercise and enriched environment did not protect against neuron loss in CA1, CA2 and CA3 areas of the hippocampus, despite a substantial increase in brain-derived neutrophic factor (BDNF) levels in dentate gyrus of the exercise and KA-treated animals. PMID- 15794995 TI - Relative levels of motivation and asymmetries of viewing and detour task in guppies (Poecilia reticulata). AB - Cerebral lateralisation once thought to be confined to humans has been reported for a range of vertebrate species now. We report here biases in visual perceptual processing in a teleost fish. Female guppy fish used the right eye preferentially to view a familiar stimulus. This bias reversed on being presented with a strange female guppy, the left eye being used more to view it. This pattern of viewing is probably associated with the right eye system, which is used to view a stimulus with an intention to approach it. The increase in the left eye use, to view a stranger may be associated with the role of the left eye in comparing the features of a strange conspecific. In the second experiment, lateralisation of viewing visual stimuli that could evoke different levels of motivation to biologically relevant stimuli was tested. It is known that female guppies prefer to approach orange coloured males. Lateralisation of detour response as well as eye use after detour to view a dull or an orange male stimulus was recorded in deprived female fish. There was a bias to detour to the left side; which was more significant for the orange than the dull male. Once the female guppies detoured the cage they preferentially used the left eye to view the male conspecific; this being significant for the deeply orange male. Thus, colouration of males evoking different levels motivation can be used to measure lateralisation in guppies. PMID- 15794996 TI - Callosal agenesis affects consistency of laterality in a paw preference task in BALB/cCF mice. AB - We tested the hypothesis that the ontogenetic development of the corpus callosum (CC) affects the consistency of laterality in a paw preference task. Adult male mice (55 normal and 29 acallosal) of the BALB/cCF strain were initially tested (twice; inter-test interval: 72 h) in an unbiased setup in which both forepaws could easily perform a reaching movement. In a subsequent test, animals were placed in a biased setup that favored the use of the non-preferred paw. Acallosal and normal mice were strongly lateralized in the unbiased setup. Additionally, while normal mice did not present a populational bias favoring one of the paws, acallosal mice presented a significant bias favoring the left paw. In the biased setup, left- and right-pawed normal mice were equally consistent (approximately 65% of the animals, in both groups, used the preferred paw of the initial two tests, in spite of the bias). Conversely, while left-pawed acallosal mice were as consistent (65%) as normal mice, only 20% of right-pawed mice were consistent. These results suggest that the development of the CC affects consistency of laterality in a side-dependent manner. These results are discussed considering the role of the CC in the establishment of behavioral lateralization. PMID- 15794997 TI - Inhibitors of carbohydrate metabolism reduce undirected song production at doses that do not alter food intake in singly housed male zebra finches. AB - Previous findings in our laboratory indicate that food availability and/or the balance of metabolic fuels may play a role in the production of undirected song in singly housed adult male zebra finches (Taeniopygia guttata). In this study, 2 deoxyglucose (2-DG) or 2,5-anhydro-d-mannitol (2,5-AM) were used to attenuate the circadian shift from lipid to carbohydrate metabolism, which normally occurs at the onset of the light phase in free-feeding, singly housed zebra finches, in order to evaluate the possibility that carbohydrate metabolism influences the production of undirected song. Food intake was also measured. Both drugs (which block carbohydrate metabolism and increase reliance on lipid metabolism) produced dose-dependent reductions in undirected singing, while food intake was not altered. Our results suggest that undirected singing (and possibly other voluntary and/or social behaviors) is sensitive to the availability of dietary fuels, whereas, food intake may show a greater regulation by the availability of stored fuels. PMID- 15794998 TI - The gradient of luminosity between open/enclosed arms, and not the absolute level of Lux, predicts the behaviour of rats in the plus maze. AB - The effect of the gradient of luminosity between the open and the enclosed arms (O/E(DeltaLux)) of the elevated plus maze (EPM), upon the level of fear/anxiety in rats submitted to the trial 1/trial 2 paradigm was investigated. Male Wistar rats were assigned to freely explore either of three EPM configuration, with the enclosed arm walls constructed with either translucent glass (O/E(DeltaLux)=11), opaque glass (O/E(DeltaLux)=96) or wood (O/E(DeltaLux)=141), for 2 consecutive days (trial 1/trial 2). Independently of the EPM configuration, rats exhibited increased fear during trial 2 relative to trial 1, thus indicating that the level of O/E(DeltaLux), at least in the range used here, is not a determinant variable for the establishment of increased anxiety induced by prior maze experience. The groups tested under 11 and 141 O/E(DeltaLux) were those who exhibited the low and higher level of open arm avoidance, respectively. There was also an increased open arms avoidance over trial 1 in rats tested under 11 and 96 O/E(DeltaLux), only. These results suggest that the enclosed arm preference of rats during trial 1 EPM procedure may be changed by the level of O/E(DeltaLux) of the test. The present results are discussed with respect to the controversy regarding the role of luminosity on EPM performance. PMID- 15794999 TI - Corticosterone treatment of the chick embryo affects light-stimulated development of the thalamofugal visual pathway. AB - By injecting a single 60 microg dose of corticosterone into the eggs of domestic chicks on day 18 of incubation, we have shown that elevated levels of this hormone affect the development of asymmetry in the visual projections from the thalamus to the Wulst regions in the left and right hemispheres of the forebrain. In vehicle-treated (control) embryos this visual pathway develops asymmetry in response to light stimulation during the final stages of incubation, when the embryo is oriented so that its left eye is occluded by its body and its right eye can be stimulated by light entering through the egg shell. Pre-hatching exposure to light leads to more projections from the left side of the thalamus to the right Wulst than from the right side of the thalamus to the left Wulst, as confirmed here by injection of the tracers Fluorogold and Rhodamine into the left and right Wulst followed by counting the number of labelled cell bodies in the thalamus (asymmetry greater in males than females). The chicks injected with corticosterone pre-hatching did not develop any group bias for asymmetry in response to light exposure before hatching. They were random with respect to presence/absence of lateralization and, when present, the lateralization was not as strong as in the controls and its direction was random. The corticosterone treated group had fewer projections from the left side of the thalamus to the right Wulst than did the controls. The results are considered with respect to maternal deposits of the hormone in the yolk and pre-hatching stress of the embryo. PMID- 15795000 TI - Anxiolytic and anxiogenic effects of kindling--role of baseline anxiety and anatomical location of the kindling electrode in response to kindling of the right and left basolateral amygdala. AB - Effects of kindling of right and left basolateral amygdala (BLA) on plus maze anxiety was studied. Using a validated retest paradigm, it was possible to retest rats in the plus maze without increasing anxiety on retest. This permitted determining prekindling baseline levels of plus maze anxiety. Right BLA kindling of high baseline anxiety rats was anxiolytic one week after kindling. Right BLA kindling of low baseline anxiety rats was anxiogenic. In addition, left BLA kindling was either anxiogenic or without effect on plus maze anxiety, depending on baseline anxiety. Effects in left BLA differ from previous work showing anxiolytic effects of left BLA kindling. The discrepancy could be explained in part by prekindling baseline anxiety. These findings require modification of the previous conclusion that left hemisphere (left BLA) kindling is anxiolytic and right BLA kindling is anxiogenic in the plus maze. Rather the hemisphere difference may be due to an interaction between baseline anxiety level and kindling. If true, anxious disposition in rodents may interact with amygdala kindling to change amygdala function differently. Kindling and baseline anxiety effects on other behaviors (such as risk assessment and resistance to capture) are also described. Present data in the light of past studies suggest both premorbid anxiety state and location of the kindling electrode contribute to the effects of kindling on behavior. PMID- 15795001 TI - Nociceptor and age specific effects of REM sleep deprivation induced hyperalgesia. AB - REM sleep deprivation (REMSD) has been shown to increase rates of negatively reinforced operant behavior, but not operant responding maintained by positive reinforcement. The reason for this differential effect is currently unknown. We hypothesize that REMSD can increase sensitivity to noxious stimuli. In the present study, we sought to determine if REMSD was associated with a change in response to noxious heat (i.e., altered nociceptive sensitivity). Two groups of rats, aged 6 and 22 months, were subjected to hotplate algesia testing at two different temperatures (44 and 52 degrees C). Initially, baseline numbers of responses and total response time were obtained at 44 degrees C. Animals then were exposed to 48 h of REMSD or control conditions. The frequency and duration of hindpaw responses (licking and guarding) increased for young animals only after REMSD and none of the control conditions. Old rats showed increased duration of nocifensive responding after REMSD and tank control conditions without a change in the number of responses at 44 degrees C. Latency to first nocifensive response was significantly longer in the 44 degrees C hotplate tests, but decreased to levels observed throughout the 52 degrees C hotplate tests following REMSD and TC conditions. These findings suggest that REMSD increases nociceptive sensitivity under conditions of sustained, selective C nociceptor activation (42 degrees C), but not under conditions of phasic A-delta activation (52 degrees C). The findings also indicate that age can be a significant variable in REMSD studies. PMID- 15795002 TI - Selective interaction of nitric oxide synthase inhibition with phencyclidine: behavioural and NMDA receptor binding studies in the rat. AB - The psychotomimetic drugs, phencyclidine (PCP) and MK-801, are non-competitive antagonists of the N-methyl-d-aspartate (NMDA) receptor and used as pharmacological tools to mimic a possible NMDA receptor hypofunction in schizophrenia. These drugs were tested in two behavioural paradigms in the present study: prepulse inhibition (PPI) of acoustic startle and locomotor activity (LMA) in an open field. Recent studies show that several behavioural and biochemical effects of PCP are blocked by nitric oxide synthase (NOS) inhibition. Hence, it is likely that some effects of PCP are mediated via an increase in NO production, an assumption not in accordance with the NMDA receptor antagonistic effect of PCP. Experiments were conducted in rats to further elucidate the involvement of NO-dependent mechanisms in the effects of PCP and MK-801, and how these effects may involve the NMDA receptor. The NOS inhibitor N(G)-nitro-l arginine methyl ester (L-NAME) (10 mg/kg) normalised the disruptive effect of PCP (2 mg/kg) on PPI and the stimulatory effect of PCP (4 mg/kg) on LMA. In contrast to these observations, the deficit in PPI induced by MK-801 (0.1 mg/kg) was not affected by L-NAME (10, 20 or 40 mg/kg). MK-801 (0.15 mg/kg)-induced hyperlocomotion was not affected by L-NAME (10 mg/kg), but attenuated by L-NAME (40 mg/kg). Furthermore, receptor binding studies aimed at investigating the influence of L-NAME on the binding of PCP to the MK-801-sensitive NMDA receptor binding site failed to show such an influence. These results suggest that the NO sensitive effects of PCP are not sufficiently explained by its antagonistic effect at the NMDA receptor channel complex. PMID- 15795003 TI - Noradrenaline involvement in the memory-enhancing effects of exposure to a complex rhythm stimulus following discriminated passive avoidance training in the young chick. AB - Previous research in our laboratory has demonstrated a significant memory enhancing effect of exposure to a complex rhythm stimulus following weakly reinforced passive avoidance learning in chicks. The aim of this study was to explore whether noradrenaline mediates this process. Chicks were trained on a strongly-reinforced single-trial passive avoidance task involving discrimination between two coloured beads. Intracerebral administration of the protein synthesis blocker, anisomycin, revealed that a phase of memory formation sensitive to arousal levels was extended by approximately 35 min following exposure to the complex rhythm stimulus. Administration of 2,4-dinitrophenol showed that this extension occurred during phase B of intermediate-term memory. Finally, a higher dose of the beta-adrenergic receptor antagonist, propranolol, was required to inhibit long-term memory in the presence of the auditory stimulus than in its absence. These findings suggest that the memory-enhancing effects of the complex rhythm stimulus may be mediated by noradrenaline, possibly via an increase in physiological arousal. PMID- 15795005 TI - Acute third ventricular administration of leptin decreases protein and fat in self-selecting rats. AB - The peripheral administration of leptin reduces food intake (FI) body weight gain (BWG) and modifies food choice. The aim of this study was to examine the effect of acute cerebral injections of leptin on food selection in rats. Male rats were first adapted to the food choice paradigm (protein, carbohydrate, fat) for 3 weeks. They were then implanted with a cannula in the third ventricle. Leptin (leptin group=L) or saline (control group=C) injections were performed at either the beginning or the end of the night at 4-day intervals. FI was recorded continuously, 3 days before, during and then after injections. Rats were sacrificed 86 h after the second injection. After both injections, BWG and FI were reduced. The reduction in FI concerned only nocturnal intake, whatever the timing of the injection. When the injection was given at the beginning of the night, the reductions after a 1-h latency period were -45% and -27.5% during the first and second days, respectively. Following the second injection, the same effects were observed immediately (-16% and -41%, respectively). Only the fat and protein intakes were significantly reduced. This lower FI was due to a reduction in meal size and duration. The reduction resulted in a lower BWG and total white adipose tissue mass. At the time of sacrifice, 6 h after food deprivation, leptinemia and insulinemia were reduced in leptin-treated rats. Glycemia values were identical. It was thus demonstrated that central leptin was a satiation factor rather than a satiety factor. PMID- 15795004 TI - Altered mesoaccumbens and nigro-striatal dopamine physiology is associated with stereotypy development in a non-rodent species. AB - Stress-induced changes in mesoaccumbens dopamine neurophysiology have been associated with the development of stereotypic behaviour in in-bred strains of laboratory rodents. This experiment evaluated whether similar changes are associated with environmentally-induced stereotypic behaviour in a higher vertebrate species, the horse. D1- and D2-like dopamine receptor densities (B(max)) and dissociation constants (K(d)) were measured in control (n=9) and stereotypy (n=9) horses in the nucleus accumbens, caudate nucleus, putamen, substantia nigra and ventral tegmentum brain regions. Results revealed that stereotypy horses had significantly higher (P<0.05) dopamine D1 and D2 receptor densities (B(max)) in the nucleus accumbens compared to non-stereotypy controls. D1 receptor densities (B(max)) and D2 receptor affinity (K(d)) were also significantly lower in the caudate nucleus brain region of stereotypy horses (P<0.05). No other significant results were observed. These results demonstrate that stereotypy horses have increased activity within the mesoaccumbens dopamine pathway and, thus, that the development of environmentally-induced stereotypy may be associated with changes in motivational systems within the animal. PMID- 15795006 TI - Procedural memory in Korsakoff's disease under different movement feedback conditions. AB - Within the field of cognitive neuroscience, it has become widely accepted to distinguish between declarative and nondeclarative memory, with different neurobiological substrates subserving these memory structures. This distinction has been inferred from the study of amnesic patients, including those suffering from Korsakoff's syndrome. It is commonly agreed that Korsakoff patients demonstrate intact memory for motor and perceptual skills (nondeclarative) whereas memory of various forms of factual knowledge (declarative) is severely impaired. In the present study, Korsakoff patients and a group of age-matched controls learned a new bimanual motor skill whereby performance was assessed in the presence and absence of augmented visual information feedback. Findings demonstrated that Korsakoff patients were able to learn and retain this skill when directive augmented information feedback was provided while no learning occurred at all in the absence of this information. These observations shed new light on the conditions required for preserved memory in amnesic patients and challenge the classic view that nondeclarative memory is invariably preserved. Instead, the quality of memory across both motor and cognitive dimensions appears to depend on the availability of task-specific information to guide performance, presumably allowing amnesic patients to bypass affected brain areas. This prompts for a reevaluation of current notions about procedural memory capacity in Korsakoff patients. PMID- 15795007 TI - Timing of conditioned responding in a nicotine locomotor conditioning preparation: manipulations of the temporal arrangement between context cues and drug administration. AB - Using a locomotor conditioning preparation, we examined whether manipulating time between exposure to distinct environmental cues and nicotine administration affected conditioned responding. Rats that received nicotine (0.42 mg/kg base) immediately before placement in an environment for 30 min on eight separate occasions displayed hyperactivity relative to controls in a subsequent injection/drug-free test. This conditioned hyperactivity was weaker if nicotine was administered 15 min before environment exposure. Conditioning was not evidenced when nicotine was administered 15 min after placement or upon removal from the environment. In a follow-up experiment, rats received 45 min in the environment; nicotine was administered 15 min after placement. This group showed conditioning that was localized to the last two-thirds of a 45 min test indicating that a 15 min delay did not prevent conditioning given 30 min of environment/nicotine overlap. This apparent timing of conditioned responding was not due to increasing environment exposure to 45 min. Further, a state-dependent environmental familiarization account of locomotor hyperactivity during testing was eliminated by the finding that rats displayed temporally specific increases in activity on the test day despite the fact that the context was previously experienced without drug for 15 min on eight consecutive days. PMID- 15795008 TI - Effects of intraperitoneal lipopolysaccharide on Morris maze performance in year old and 2-month-old female C57BL/6J mice. AB - Several studies have shown that systemic lipopolysaccharide (LPS) or interleukin 1beta (IL-1beta) may affect performance in various learning tasks, including the Morris water maze. In the current study, female C57BL/6J mice, either 2 months or 1 year of age, were given 5 days of testing followed by 3 days of rest, and then three additional days of testing. Mice either received a single LPS injection on day 1 and saline on days 2-5, LPS injections on days 1-5, or saline injections on days 1-5. Daily LPS administration significantly prolonged latency for the animals to find the platform, and decreased their swimming speed. Year-old mice treated with LPS each day also exhibited significantly higher levels of thigmotaxis in the maze. Despite effects on latency and swim speed, no effect of LPS treatment was observed for distance traveled to the platform or other measures that clearly indicate disruption of learning in the maze. On the other hand, age was a significant factor affecting both latency and distance, with older animals swimming greater distances to find the platform. Additionally, older animals were more adversely affected by daily LPS treatment. In this study, although LPS-induced performance impairments in the Morris water maze were noted, particularly in older animals, these effects were not clearly indicative of learning impairment per se. PMID- 15795009 TI - Comparison of 6-hydroxydopamine-induced medial forebrain bundle and nigrostriatal terminal lesions in a lateralised nose-poking task in rats. AB - The nigrostriatal degeneration underlying Parkinson's disease (PD) is commonly modeled in experimental animals by injection of the neurotoxin 6-hydroxydopamine (6-OHDA). Although a wide variety of simple behavioural screens exist to assess the impact of such dopamine lesions, more complex tasks that assess multiple parameters of an animal's performance may provide a more sensitive measure of the resulting functional impairment. This study assessed the performance of two unilateral lesion models of PD in a lateralised nose-poking task in the nine-hole box test apparatus. This task assesses the accuracy and speed of movements to either side of a rats' head, as well as a number of errors of performance. Rats with complete unilateral dopamine depletion (induced by injection of 6-OHDA into the medial forebrain bundle (MFB)) attempted fewer trials and committed more procedural errors than controls. They developed a marked ipsilateral responding bias, with a reduced accuracy for contralateral stimuli. They were also slower to react to contralateral stimuli and to complete movements bilaterally. Rats with unilateral nigrostriatal terminal lesions (induced by multiple injections of 6 OHDA in the striatum) developed a similar pattern of deficits, but they were significantly less impaired and spontaneously recovered to pre-operative levels by 4 months post-lesion. This experiment confirms that the lateralised nose poking task may be a powerful tool for assessment of the nature of deficit and recovery in rats with complete but not partial unilateral dopamine lesions. PMID- 15795010 TI - Systemic administration of MK-801, a non-competitive NMDA-receptor antagonist, elicits a behavioural deficit of rats in the Active Allothetic Place Avoidance (AAPA) task irrespectively of their intact spatial pretraining. AB - Spatial orientation is considered to be an animal model of human cognitive functions. Efficient navigation is believed to require a brain representation of the environment. The role of NMDA-receptor-dependent neurotransmission in encoding spatial representations has been intensively studied; however, its involvement in organizing spatial information into neural representations is poorly understood. We tested the effect of NMDA-receptor blockade on the performance of rats in Active Allothetic Place Avoidance (AAPA), a cognitive task which requires rats to separate two conflicting spatial reference frames (room and arena frames), when only the room frame is relevant for solution of the task. The NMDA-receptor blockade was achieved by systemic administration of either 0.15 mg/kg or 0.2 mg/kg of dizocilpine (MK-801), a non-competitive NMDA-receptor antagonist. Two distinct AAPA setups located in different rooms were used. The rooms had different layouts of robust extramaze landmarks. Intact rats were trained in the upstairs arena and, subsequently, they received saline or MK-801 and their performance was tested (reinforced retention). Re-acquisition of AAPA task in the downstairs room was then tested under saline or MK-801. MK-801 at the dose of 0.2 mg/kg was found to disrupt the reinforced retention of the AAPA. Re acquisition of AAPA in the novel environment was impaired by both doses of MK 801. Taken together, administration of MK-801 causes an orientation deficit in the AAPA task irrespectively of the rats' prior intact pretraining. These results support the hypothesis that NMDA-receptors play a pivotal role in cognitive processes. PMID- 15795012 TI - Farm animal networks: unraveling the contact structure of the British sheep population. AB - The spatial and temporal dynamics of many farm animal diseases depend both on disease specific parameters and on the underlying contact structure between farms. Whilst many models for farm animal diseases focus on obtaining and estimating disease transmission parameters, relatively little attention has been given to modelling the underlying network of contacts. In this paper, we present an initial analysis of two relations underlying the contact network of individual sheep breeds in Great Britain. The first relation is based on geographical proximity and the second is based on attendance at agricultural shows. These relations are combined to give a risk-potential network that is based on these two levels of interaction. The structure of each network is investigated using techniques developed in graph theory and social network analysis. PMID- 15795013 TI - Diagnosing diagnostic tests: evaluating the assumptions underlying the estimation of sensitivity and specificity in the absence of a gold standard. AB - Latent class analysis to assess the sensitivity and specificity of a diagnostic test can be carried out under different assumptions. An often applied set of assumptions is known as the Hui-Walter paradigm, which essentially states that: (i) the population is divided into two or more populations in which two or more tests are evaluated under assumption that (ii) sensitivity and specificity of the tests are the same in all populations; and (iii) the tests are conditionally independent given the disease status. This study explores the implications of these assumptions. Through simulation studies, it is shown how the size of the difference between disease prevalences within the populations influences the precision of the estimates. It is also illustrated by a simulation study how a difference in a test sensitivity between populations may result in estimates that are biased towards the sensitivity of the test in the population with highest disease prevalence, since that population estimate is supported by most of the data. It is shown that the assumption of conditional independence between tests in general cannot be ignored in latent class models. Failure to impose conditional independence will result in a model that lacks identifiability in a way that cannot be handled by adding more tests or dividing the sample into more populations. PMID- 15795015 TI - The impact of different housing systems on health and welfare of grower and finisher pigs in Switzerland. AB - The objective of the study was to evaluate the impact of housing systems on the health and welfare of grower and finisher pigs. Forty-seven "animal friendly" farms meeting Swiss criteria and participating in a Swiss subsidy program and 37 farms using traditional housing with slatted floors were evaluated in an epidemiological field study. Group-health evaluations and individual clinical examinations were performed during four visits to each farm. Evaluation of pig behaviours associated with health and welfare were assessed through indicators including presence of lesions on the snout, ears, shoulders, legs and tail. General group-health evaluations included lameness, respiratory disease, diarrhoea, ear biting, skin abnormalities, injuries, abscesses, sunburn, recumbency, ill-thrift and behavioural abnormalities such as dog sitting and tail biting. Data on management practices, medical treatment, mortality and results of slaughter inspections were also collected, and the type of housing described. Pooled faecal samples were collected and examined for intestinal parasites. Statistical analyses included descriptive statistics of indicators of health and welfare and logistic-regression analysis accounting for group effect using generalised estimation equation (GEE). Findings indicated a high health status of pigs in Swiss finishing farms. Data from the individual clinical examination were more sensitive compared to the group-health evaluations. Both the individual clinical examination and group-health evaluation indicated that "animal friendly" housing had positive effects on health and welfare of fattening pigs. PMID- 15795014 TI - Pseudorabies virus infections in wild boar: data visualisation as an aid to understanding disease dynamics. AB - Spatio-temporal survey data often form the main decision support in disease management. Consequently, disease managers rely on adequate description of the data supplied from epidemiological analysis. Practically speaking, such a description comprises of a visualisation, a deduced hypothesis and a statistical evaluation of the hypothesis. Prior to the application of traditional methods of data visualisation and hence data description, an aggregation of the data is required. This paper illustrates the resulting uncertainty in hypotheses that can be generated by traditional analytical techniques. Additionally, a method of avoiding this typical drawback in traditional data exploration is demonstrated: This is the use of data movies. Data from a 10-year study on the spread of pseudorabies virus (PrV) infections in wild boar populations were revisited. The visualisation of the data using a data movie instead of using traditional approaches for the first time enabled immediate observation of spreading infection in time and space. This dynamic perception of the spread drastically changes the understanding of the epidemiology of the virus. The paper demonstrates how data movies can account for the complete information contained within the data set. It is concluded that data movies can provide a more appropriate starting point. We recommend the data movie approach as an important addition in the epidemiological toolbox, because data movies can easily be integrated into the usual demonstration and publication process. PMID- 15795016 TI - Where should the effort be put to reduce the Salmonella prevalence in the slaughtered swine carcass effectively? AB - Infection with Salmonella Typhimurium is seldom associated with clinical disease in pigs. However, control is important as the public is concerned about the human health impact. The producers, the abattoirs and the authorities are interested in implementing procedures to mitigate this risk. To evaluate the effect of different procedures, a stochastic risk model was developed to simulate the prevalence of Salmonella infection during the production process from the live pig on the farm, to the final carcass. This paper describes the model and findings of simulating different control scenarios. The variables with maximum effect on the Salmonella prevalence on the final carcass were (1) number of herds with a high prevalence of Salmonella, (2) singeing efficiency, (3) contamination and cross-contamination at degutting and (4) cross-contamination during handling. However, improvement of any single factor in isolation had a limited impact upon the level of contamination. The largest reduction was observed when several factors were improved concurrently. PMID- 15795017 TI - All bathwater and no baby: revisiting a national curriculum and state examination for nursing? PMID- 15795018 TI - What we know and what they do: nursing students' experiences of improvement knowledge in clinical practice. AB - Nations around the world face mounting problems in health care, including rising costs, challenges to accessing services, and wide variations in safety and quality. Several reports and surveys have clearly demonstrated that adverse events and errors pose serious threats to patient safety. It has become obvious that future health professionals will need to address such problems in the quality of patient care. This article discuss a research study examining improvement knowledge in clinical practice as experienced by nursing students with respect to a patient-centred perspective, knowledge of health-care processes, the handling of adverse events, cross-professional collaboration, and the development of new knowledge. Six focus groups were conducted, comprising a total of 27 second-year students. The resulting discourses were recorded, coded and analysed. The findings indicate a deficiency in improvement knowledge in clinical practice, and a gap between what students learn about patient care and what they observe. In addition the findings suggest that there is a need to change the culture in health care and health professional education, and to develop learning models that encourage reflection, openness, and scrutiny of underlying individual and organizational values and assumptions in health care. PMID- 15795019 TI - Issues in helping students from other cultures. AB - This paper reviews some of the research highlighting evidence of problems associated with studying overseas. Issues such as problems with daily living, being in the classroom and dealing with differences in the academic system are identified. The issue of studying in another culture is considered and problems of differences of perceptions between teachers and students are noted. The paper closes with some suggestions of how teachers in universities might improve their skills in helping such students. Given that the paper has to have a 'position', the assumption is that the students, in question, are coming to 'western' countries to study. The principles also apply in many other contexts and the reader must make this adjustment, as necessary. PMID- 15795020 TI - Learning from success: improving undergraduate education through understanding the capabilities of successful nurse graduates. AB - This paper reports on a study of nursing graduates identified as high performers by their nursing unit managers. The study involved 17 graduates from two teaching hospitals, one in the inner city of Sydney and the other in regional New South Wales. The aim of this study was to identify the capabilities that were seen to be most important for successful nursing practice during the first 2-6 years following employment as registered nurses and to evaluate through backward mapping, the degree to which university programmes were developing these capabilities. This study is part of a larger scale study of a number of professional disciplines that will be used to shape the ongoing evolution of the undergraduate programmes in these disciplines at the University of Technology, Sydney (UTS). The results from this study provide evidence that while capability in technical skill is necessary for successful practice as a nurse, it is certainly not sufficient. A range of 'emotional intelligence' capabilities have been identified by both graduate nurses and their nursing unit managers as being significant factors for successful practice. It would be important that a curriculum provided a range of learning experiences that included content related to emotional intelligence, and it may be that by directing attention to the total undergraduate learning experience rather than just what is taught, that curriculum developers can do much to provide educational opportunities that encourage the capabilities identified. PMID- 15795021 TI - The experiences of a group of pre-registration mental health nursing students. AB - Research that only utilises mental health students as a sample group is quite rare. Yet there are many potential stressors that may impact on the experience of these students including; the theory-practice gap, the learning culture experienced in clinical practice, the effects of the programme's assessment and learning strategies and the intentional and unintentional effects of the curriculum. Therefore the aim of this study was to explore the experiences of a group of mental health student nurses by utilising these four concepts as a framework. Focus groups were used to generate data and the sample consisted of fourteen mental health branch students who, within the pre-registration nursing programme, formed a problem-based learning group. The group met on an annual basis at key junctures during the three-year diploma programme. The focus groups were tape-recorded and the data interpreted and organised by utilising an adapted form of content analysis. Three themes were identified; the conflicting pressures of life as a mental health student nurse, adjustment to problem based learning and experiences in mental health practice and views on mentorship. A discussion of the implications of the findings for the planning of nursing curricula and mental health branch programmes is outlined. PMID- 15795022 TI - Nursing education in Slovenia and its impact on nurses publishing in their professional journal. AB - The paper describes development of nursing education in Slovenia (from Associate Degree to Diploma level, with BSN programme of brief duration) and discusses its impact on structure and contents of the Journal of the Slovenian Nursing Association (JSNA). Extensive bibliometric analysis was performed on all scientific and technical papers published in the JSNA between 1976 and 2001. Statistical analyses were carried out to study characteristics and inter relations of authorship, article type, cited references and subject classification. A positive impact of improved and extended education of nurses on quantity and quality of their publications was confirmed. Nevertheless, further steps in terms of education and research are needed fully to establish nursing as an independent profession in Slovenia, and there is still much room for improving the standard of the articles in the JSNA. PMID- 15795023 TI - Anti-racist innovation and nurse education. AB - The focus on institutional racism within the Higher Education (HE) sector in the United Kingdom and nurse education, in particular, has so far been seriously lacking in investigation and scrutiny. The Race Relations (Amendment) Act (RRAA2000) has pushed institutional racism to the forefront of debates in public services, including both education and health services. This paper seeks to operationalise some key aspects of the debate over institutional racism and relates it to both these sectors. Based on empirical work funded by the Higher Education Funding Council for England and Wales (HEFCE) Innovations Project, this paper offers nurse education a framework to not only comply with the legal requirement of 'promoting racial harmony' (RRAA 2000) and the expectations from nursing as enshrined in the Code of Professional Conduct (NMC, 2000. The Code of Professional Conduct: Protecting the Public Through Professional Standards. NMC, London), but goes further to consider some key questions for anti-racist interventions. PMID- 15795024 TI - Cultural competence among nursing students and faculty. AB - Healthcare consumers are entitled to culturally competent care. Therefore, nursing curricula need to include cultural content and student nurses and faculty members need to be culturally competent. The purpose of the study was to describe cultural competence of students and faculty at a college of nursing and to discuss the implications for nursing curricula related to cultural competence. Campinha-Bacote's model (Campinha-Bacote, J., 1994. Cultural competence in psychiatric mental health nursing. Nursing Clinics of North America 29 (1), 1-8.) of culturally competent care provided the theoretical framework. The Inventory for Assessing the Process of Cultural Competence (IAPCC) (Campinha-Bacote, J., 1998. The Process of Cultural Competence in the Delivery of Healthcare Services: A Culturally Competent Model of Care. Transcultural C.A.R.E. Associates, Cincinnati, OH. Available from: .) was used to measure levels of self-reported cultural competence. A convenience sample of 88 first year, 121 fourth year baccalaureate students and 51 faculty members at a college of nursing was studied. Analysis of variance revealed a statistically significant difference (F=43.915, df=259, p<.0001) between the three groups. A positive correlation was found between IAPCC scores and several demographic variables. Findings suggest that cultural competence can be increased by including structured cultural content in nursing curricula. PMID- 15795025 TI - The changing landscape of doctoral education: Introducing the professional doctorate for nurses. AB - The last decade has heralded the introduction of an alternative form of doctoral education for nurses in the United Kingdom, the professional or taught doctorate. First introduced in 1995 in the UK the number of professional doctorates for nurses has steadily increased totalling more than 23 programmes in 2004, a trend that seems set to continue. This paper presents those factors leading to the introduction of the professional doctorate in mainstream higher education generally and those leading to its adoption by the profession nursing. Professional doctorates are defined variously and these are considered relative to the traditional PhD. It will be some time before the full benefits of these programmes are realised and an empirical basis established. Meantime this paper highlights some of the potential benefits and some concerns whilst advocating recommendations that include the longitudinal evaluation of such programmes. PMID- 15795026 TI - The mature student question. AB - Demographic changes resulting in an increase of mature students on to pre registration nursing programmes within our university has lead to the development of a welcoming programme for pre-registration mature nursing students. Three questionnaires were utilised in the development and evaluation of the welcoming programme. Each questionnaire had open ended questions which were thematically collated and summarised. The closed questions were computer analysed to derive simple descriptive statistics. The main components of the welcoming programme were socialisation of the individual to university life and study skills. The study found that there was a small significant difference between the needs of mature students and of standard entry students in relation to information about the nursing course, study skills and time management. The students who participated in the welcoming programme progressed a little better on the nursing course. Students' recommendations for further development of the programme included, additional study and IT skills and an introduction to subject areas on the nursing course. If we are to continue to recruit mature students on to our nursing courses we need to ensure that supports are in place to facilitate the mature student prior to and during their nursing course. PMID- 15795027 TI - Cooperative learning and case study: does the combination improve students' perception of problem-solving and decision making skills? AB - The purpose of this study was to investigate the effectiveness of cooperative learning techniques combined with case study on nursing students' self-perception of problem-solving and decision making skills in comparison with other teaching learning methods. A quasi-experimental pre-test to post-test static group comparison design was used to determine the influence of the different teaching methodologies. Three associate degree colleges provided a convenience sample of nursing students (N=123). An experimental group (n=31) was taught through cooperative learning and case study. One comparison group, was taught through lecture and large group case study (n=46) and another comparison group was taught through lecture only (n=24). A third comparison group taught through lecture and occasional use of non-cooperative learning groups using continuing case study (n=22) was used as a post-test only control group. No two groups were significantly different at the 0.5 level. The results, however, showed that the experimental group obtained scores indicating somewhat better self-perception of both problem-solving and decision making skills. PMID- 15795029 TI - Applying radioecology in a world of multiple contaminants. AB - This paper presents a review of the current status of radioecology and how it might be applied to contribute to a broader range of environmental pollution and contamination problems. In many respects radioecology is a unique and specialised branch of environmental science. However, radioecologists could be much more aware of the similarities and differences between their field and other fields of environmental pollution science. Areas of common interest described herein are exposure and risk assessment, and the problem of the bioavailability of potentially toxic substances in environmental media. It is concluded that radiometric methods and radioecological modelling methods can assist significantly with understanding and quantifying both these issues. The existence of multiple contaminants at many locations around the world dictates that assessment and remediation exercises should be carried out in an integrated fashion, as a partnership between scientific disciplines. PMID- 15795030 TI - Fibre crops as alternative land use for radioactively contaminated arable land. AB - The transfer of radiocaesium, one of the most important and widespread contaminants following a nuclear accident, to the fibre crops hemp (Cannabis sativa L.) and flax (Linum usitatissimum L.) as well as the distribution of radiocaesium during crop conversion were studied for sandy soil under greenhouse and lysimeters conditions. Soil parameters did not unequivoqually explain the transfer factors (TF) observed. TFs to flax stems ranged from 1.34 to 2.80x10(-3) m2 kg(-1). TFs to seeds are about a factor of 4 lower. During the retting process for separating the fibres from the straw, more than 95% of the activity was removed with the retting water. For hemp, the TF to the stem was about 0.6x10(-3) m2 kg(-1). For hemp, straw and fibres were mechanically separated and TF to straw was about 0.5x10(-3) m2 kg(-1) and to fibres 1.0x10(-3) m2 kg(-1). Generally, the TFs to the useable plant parts both for hemp and flax, are low enough to allow for the production of clean end-products (fibre, seed oil, biofuel) even on heavily contaminated land. Given the considerable decontamination during retting, contamination levels in flax fibres would only exceed the exemption limits for fibre use after production in extreme contamination scenarios (>12,300kBq m(-2)). Since hemp fibres are mechanically separated, use of hemp fibres is more restricted (contamination <740kBq m(-2)). Use of stems as biofuel is restricted to areas with contamination levels of <250 and 1050kBq m(-2) for flax and hemp, respectively. Use of seeds for edible oil production and flour is possible almost without restriction for flax but due to the high TFs to seed observed for hemp (up to 3x10(-3) m2 kg(-1)) consumption of hemp seed products should be considered with care. PMID- 15795031 TI - Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays. AB - Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137Cs pollution. PMID- 15795032 TI - Soil migration and plant uptake of technetium from a fluctuating water table. AB - Soil columns (50x15 cm) were used to determine the potential for 95mTc (as a surrogate for 99Tc which is an important component of some radioactive waste) to migrate from a contaminated, fluctuating water table, through sandy loam soil and into perennial ryegrass. Upward migration was significantly retarded with, generally, only the bottom few centimetres of soil becoming contaminated over the 6 months of the experiment. This is thought to have been due to the presence of anoxic conditions within the water table leading to the reduction of pertechnetate to Tc(IV) species which are relatively insoluble. However, some evidence of very slow upward migration over time was found. Only a small and inconsistent transfer of activity into the perennial ryegrass was observed. Whilst these observations would suggest that 99Tc is less important than radionuclides such as 129I and 36Cl in terms of the risk associated with radioactive waste disposal, the potential for a slow upward migration, and/or a pulse-release following the re-oxidation of reduced soil in which 99Tc has accumulated should not be overlooked. PMID- 15795033 TI - Evolution of Sr distribution coefficient as a function of time, incubation conditions and measurement technique. AB - A thorough understanding of the dynamics of radiostrontium in soil is required to allow accurate long-term predictions of its mobility. We have followed the soil solution distribution of 85Sr as a function of time under controlled conditions over 4 months and studied the effect of soil moisture content and organic matter amendments. Data have been compared to redox conditions and soil pH. To fuel the ongoing debate on the validity of distribution coefficient (K(d)) values measured in dilute suspension, we have compared values obtained from the activity concentration in soil solution obtained by centrifugation to data obtained in suspension with or without air-drying of the soil samples after incubation. The 85Sr adsorption properties of soil, incubated without prior contamination were also measured. There is some time-dependent adsorption of Sr. This is partly due to changing soil composition due to the decomposition of added organic matter and anaerobic conditions induced by flooding. There is also a kinetic effect, but adsorption remains largely reversible. Most of the observed effects are lost when soil is suspended in electrolyte solution. PMID- 15795034 TI - Modelling 137Cs uptake in plants from undisturbed soil monoliths. AB - A model predicting 137Cs uptake in plants was applied on data from artificially contaminated lysimeters. The lysimeter data involve three different crops (beans, ryegrass and lettuce) grown on five different soils between 3 and 5 years after contamination and where soil solution composition was monitored. The mechanistic model predicts plant uptake of 137Cs from soil solution composition. Predicted K concentrations in the rhizosphere were up to 50-fold below that in the bulk soil solution whereas corresponding 137Cs concentration gradients were always less pronounced. Predictions of crop 137Cs content based on rhizosphere soil solution compositions were generally closer to observations than those based on bulk soil solution composition. The model explained 17% (beans) to 91% (lettuce) of the variation in 137Cs activity concentrations in the plants. The model failed to predict the 137Cs activity concentration in ryegrass where uptake of the 5-year old 137Cs from 3 soils was about 40-fold larger than predicted. The model generally underpredicted crop 137Cs concentrations at soil solution K concentration below about 1.0 mM. It is concluded that 137Cs uptake can be predicted from the soil solution composition at adequate K nutrition but that significant uncertainties remain when soil solution K is below 1 mM. PMID- 15795035 TI - Uranium distribution and cycling in Scots pine (Pinus sylvestris L.) growing on a revegetated U-mining heap. AB - We determined the uranium distribution in soil and its allocation in compartments of 35-year-old Scots pine developed on a revegetated U-mining heap. The processes controlling the dynamics of U recycling were identified and further quantified in terms of annual fluxes. As pine developed, an acid humus layer emerged leading to weathering of the alkaline mining debris but this had little effect on U mobility in the soil profile. Increased U mobility mainly involved a translocation of U to metal-humus chelates in surface layers. The root compartment accounted for 99.3% of the U budget in tree, thus serving as an effective barrier which restricts U uptake. The current root uptake and transfer of U to upper parts of the tree amounted to about 3g ha(-1) y(-1), i.e. less than 0.03% of the current NH4 exchangeable U pool in the soil (0-30 cm). Allocation and translocation pattern made it clear that a dominant fraction of the translocated U moves passively with the ascent xylem sap, most likely as a soluble complex, and steadily accumulates in the needles. Consequently, 97% of the U annual uptake is returned to the soil through litterfall. At the studied site, the risk of U dissemination due to biomass turnover or trunk harvest was low when considered in relation to the current "exemption level" for U. PMID- 15795037 TI - Effect of K and bentonite additions on Cs-transfer to ryegrass. AB - Bentonite amendments are generally ineffective in reducing the soil-to-plant radiocaesium transfer but have previously been shown that bentonites in the K form having been subjected to wetting-drying cycles had pronounced radiocaesium binding capacities. We have investigated the effect of wetting-drying (WD) on Radiocaesium Interception Potential (RIP) development in three K-bentonites and K bentonite soil mixtures, using a variety of procedures: homogenisation of the bentonites with K through dialysis (K(B)), or partial transformation of the bentonite to the K-form in the presence of a solution of K2CO3 (K(L)) or in presence of solid K2CO3 (K(S)). Of the three strategies tested, addition of K2CO3 (solid) at a dose of 2 meq g(-1) clay and adding the K-bentonite mixtures to the soil resulted in the highest RIP increase after 20 WD cycles. The procedure giving the highest RIP yield is the most practical for further applications and was used in a pot experiment under greenhouse condition. When expressing the RIP increase of the soil-bentonite mixtures per unit bentonite added (RIP yield), 28- to 110-fold RIP increases were observed up to a value of approximately 60,000 meq kg(-1) (6 times higher than the RIP for illite). The beneficial effect following K-bentonite application was shown to be dependent both on a sorption enhancement effect (direct RIP effect) and fixation effects (indirect RIP effect). Greenhouse testing proved that the RIP effects observed in greenhouse could be predicted by making use of the sorption data from the laboratory tests. Optimum soil-amendment would be obtained with bentonites with high initial sorption RIP and a high sorption RIP increase when subjected to WD in the presence of potassium. Hypothised Transfer Factor (TF)-reductions of at least 10-fold could result when mixing approximately 1% bentonite, like Otay bentonite (RIP yield 99,000 meq kg( 1) after WD in presence of K if only fine particle size of <1mm considered) with the contaminated ploughing layer. PMID- 15795038 TI - Can barium and strontium be used as tracers for radium in soil-plant transfer studies? AB - Radium is one of the prominent potential contaminants linked with industries extracting or processing material containing naturally occurring radionuclides. In this study we investigate if 133Ba and 85Sr can be used as tracers for predicting 226Ra soil-to-plant transfer. Three soil types were artificially contaminated with these radionuclides and transfer to ryegrass and clover was studied. Barium is considered a better tracer for radium than strontium, given the significant linear correlation found between the Ra and Ba-TF. For strontium, no such correlation was found. The relationship between soil characteristics and transfer factors was investigated. Cation exchange capacity, exchangeable Ca+Mg content and soil pH did not seem to influence Ra, Ba or Sr uptake in any clear way. A significant relation (negative power function) was found between the bivalent (Ca+Mg) concentration in the soil solution and the Ra-TF. A similar dependency was found for the Sr and Ba-TF, although less significant. PMID- 15795036 TI - Uptake and distribution of 90Sr and stable Sr in rice plants. AB - The stable Sr content in the aboveground parts of rice plants at various growth stages, and the distributions of 90Sr and stable Sr in rice plant components, such as polished rice, rice bran, hull, straw and root, at harvest time, were determined. The total Sr content in the aboveground rice plants was dependent on the growth stage and followed the sigmoidal shape of the growth curve. The concentration of 90Sr among the different components of rice plants varied within two orders of magnitude, whereas the 90Sr/Sr concentration ratio had a constant value. Therefore, the translocation rate of 90Sr in rice plants had similar values to that of stable Sr. However, the 90Sr/Sr concentration ratio for the rice plants was different for each study site. Only 0.6% of the total Sr was found in polished rice, while more than 99% was found in the non-edible components, of which 87% was present in the straw. These findings suggest that 90Sr in the non-edible parts could have been transferred to humans through the soil-plant system and/or feed-livestock pathway. The soil-to-plant transfer factor of 90Sr in polished rice was 0.0021 +/- 0.00007, which was two orders of magnitude lower than that in the straw. The percentage of 90Sr removed from the upper soil layer to the aboveground biomass of rice plants at harvest time was calculated as 0.094%. It is possible that approximately 0.1% of the total 90Sr content in the surface soil layer is removed from the soil-plant system by human activities every year. PMID- 15795039 TI - Bioavailability of radiostrontium in soil: experimental study and modeling. AB - Parameters related to 90Sr mobility in the soil-plant system are reported: exchangeable content, selectivity coefficient, and transfer factor. Large mobility of 90Sr in different soil types was shown. The fraction of exchangeable 90Sr varied between 70 and 90%. The selectivity coefficient K(C)(90Sr/Ca) values were in the range 1.3-2.5. The radionuclide transfer factors (TF) varied by a factor of 9.6 for barley seedlings and by a factor of 6.6 for lupine seedlings. The exchangeable Ca content was the determinant soil parameter responsible for differences in 90Sr biological availability. A static model was devised that describes 90Sr sorption from soil solution by soil and on the root surface. The parameter of 90Sr bioavailability (A) has been suggested. Parameter A was calculated from data on soil exchangeable Ca content and 90Sr mobility indicators -exchangeable fraction of the radionuclide and the selectivity coefficient K(C)(90Sr/Ca). A correlation was found between TF and parameter A. PMID- 15795040 TI - Thorium uptake by wheat at different stages of plant growth. AB - Data on biogeochemistry of thorium are rather limited. So far little is known about toxic effects of small amounts of the radionuclide on higher plants. In this study the uptake of thorium by wheat seedlings was measured by greenhouse experiments. Germination of wheat seeds for 6 days in the presence of thorium resulted in accumulation of the metal in all parts of the seedlings. When the Th rich seedlings were transferred to normal soil and were grown there further for 7 days, Th concentrations in roots and leaves decreased significantly (in leaves the Th content decreased up to the level of Th in the control plants). In seeds, however, Th content remained unchanged. An increase of Th content in roots and seeds was also observed as a result of addition of thorium to soil but in this case the concentration of Th in leaves did not change. The accumulation of Th in plants affected the uptake of other elements including essential macro-nutrients. The most strongly affected part of the plants was leaf. PMID- 15795041 TI - Distribution of 90Sr and 137Cs in Arctic soil profiles polluted by heavy metals. AB - Effects of industrial pollution on the behaviour of radionuclides in spruce forest ecosystems were studied along a gradient from of a copper-nickel smelter in Monchegorsk, NW Russia. A reference site was situated in Lapland, Finland, 152 km west of Monchegorsk. Most of the total 137Cs activity in soil was in mineral (E and B) horizons, except at the reference site where the major part was still in the organic surface layer. Most of the total 90Sr activity still remaining in the soil profile was found in the surface layer, but the relative amount decreased with increasing level of industrial pollution. Pollutants from the smelter clearly affected the chemical speciation of radionuclides. Smaller amounts of exchangeable radionuclides were present in the organic surface layer at the most polluted sites. The decline of 137Cs with decreasing distance from the smelter correlated strongly with a similar depletion in exchangeable K and Mg. Total concentrations of 137Cs and 90Sr showed high correlations with exchangeable cations, particularly in the E and upper B horizon. A sudden change in behaviour of 137Cs in the lower B horizon may be associated with changes in clay mineralogy along the soil profile caused by weathering. PMID- 15795042 TI - Observed half-lives of 3H, 90Sr and 137Cs in hydrosphere in the Vltava River basin (Bohemia). AB - A near field study was set up to follow the effects of the Temelin nuclear power plant construction. Reference levels of artificial radionuclides were monitored in the Vltava River upper course and its tributaries in the period 1990-2001. Monitoring continued even after the waste water release startup during the pilot operation in 2002. The assessment of the (90)Sr and (137)Cs concentrations histories in ground water, river bottom sediments and fish showed a decreasing trend. This trend was not influenced by the nuclear power plant pilot operation. In the case of tritium, trend of increasing concentration had been already observed since the pilot operation startup. The monitoring of changes in concentrations of artificial and natural radionuclides in influenced and uninfluenced profiles will be maintained to assess the possible influence of the operation of the Temelin nuclear power plant. PMID- 15795043 TI - Pharmaceutical and other commercial uses of the dog model. PMID- 15795044 TI - A comparison of egocentric and allocentric age-dependent spatial learning in the beagle dog. AB - Spatial discriminations can be performed using either egocentric information based on body position or allocentric information based on the position of landmarks in the environment. Beagle dogs ranging from 2 to 16 years of age were tested for their ability to learn a novel egocentric spatial discrimination task that used two identical blocks paired in three possible spatial positions (i.e. left, center and right). Dogs were rewarded for responding to an object furthest to either their left or right side. Therefore, when the center location was used, it was correct on half of the trials and incorrect on the other half. Upon successful acquisition of the task, the reward contingencies were reversed, and the dogs were rewarded for responding to the opposite side. A subset of dogs was also tested on an allocentric spatial discrimination task, landmark discrimination. Egocentric spatial reversal learning and allocentric discrimination learning both showed a significant age-dependent decline, while initial egocentric learning appeared to be age-insensitive. Intra-subject correlation analyses revealed a significant relationship between egocentric reversal learning and allocentric learning. However, the correlation only accounted for a small proportion of the variance, suggesting that although there might be some common mechanism underlying acquisition of the two tasks, additional unique neural substrates were involved depending on whether allocentric or egocentric spatial information processing was required. PMID- 15795045 TI - An investigation of the relationship between response latency across several cognitive tasks in the beagle dog. AB - Response latencies (RLs) extracted from simple motor tasks are a commonly used index of human intelligence. To date few human or animal studies have investigated the relationship between an individuals RL scores across a number of diverse cognitive tasks: Does RL remain consistent between individuals across several cognitive domains? Thus, the current study examined how RL measures gathered from beagle dogs (n=13) tested on three different cognitive tasks were related. RL scores were collected following both discrimination and reversal learning and a test of visuospatial memory, the 3 component delayed non-matching to position (3-DNMP) task. RL scores were recorded from the time the choice stimulus was presented until the animal selected a response. Results indicated that strong correlations emerged between 3-DNMP RLs and both the discrimination and reversal RLs, indicating that animals that responded fast on the 3-DNMP task also responded fast on the discrimination and reversal tasks. Interestingly, 3 DNMP RLs were more strongly correlated with reversal learning RLs. Finally, when mean RL performance across the three tasks was examined, strong RL differences emerged indicating that animals displayed significantly slower RLs on the 3-DNMP task than on the discrimination task, while reversal RLs remained indistinguishable from both. In conclusion, RLs show high between task correlations, indicating individual differences, and also vary between tasks, probably because of differences in task difficulty. These results further validate the use of RLs as an index of cognition, and also highlight the importance of further studies using animal models. PMID- 15795046 TI - A novel method for assessing contrast sensitivity in the beagle dog is sensitive to age and an antioxidant enriched food. AB - Tasks requiring visual discrimination are commonly used in assessment of canine cognitive function. However, little is known about canine visual processing, and virtually nothing is known about the effects of age on canine visual function. This study describes a novel behavioural method developed to assess one aspect of canine visual function, namely contrast sensitivity. Four age groups (young, middle aged, old, and senior) were studied. We also included a group of middle aged to old animals that had been maintained for at least 4 years on a specially formulated food containing a broad spectrum of antioxidants and mitochondrial cofactors. Performance of this group was compared with a group in the same age range maintained on a control diet. In the first phase, all animals were trained to discriminate between two high contrast shapes. In the second phase, contrast was progressively reduced by increasing the luminance of the shapes. Performance decreased as a function of age, but the differences did not achieve statistical significance, possibly because of a small sample size in the young group. All age groups were able to acquire the initial discrimination, although the two older age groups showed slower learning. Errors increased with decreasing contrast with the maximal number of errors for the 1% contrast shape. Also, all animals on the antioxidant diet learned the task and had significantly fewer errors at the high contrast compared with the animals on the control diet. The initial results suggest that contrast sensitivity deteriorates with age in the canine while form perception is largely unaffected by age. PMID- 15795047 TI - A longitudinal study of brain morphometrics using serial magnetic resonance imaging analysis in a canine model of aging. AB - Longitudinal changes in cortical atrophy, ventricular enlargement, and lesion development in serial MRI scans collected from 47 healthy dogs from 1999 (8-11 years old) to 2002 (11-14 years old) were studied. The first method involved manual region of interest volumetric analysis to examine changes in cerebral and ventricular volume during the three years. No change in cerebral volume was detected but ventricular volume increased significantly each year in 2000, 2001, and 2002. Increased ventricular volume parallels early studies of age-dependent ventricular enlargement in the brain of aging beagle dogs. The second method involved a visual analysis of co-registered serial MRIs for each subject. Consistent with the volumetric results, there was no visible change in cortical thickness indicating no cerebral atrophy, but a significant increase in ventricular size was noted. Visual examination also revealed a significant increase in number of dogs who developed aging lesions over the last 2 years in 2001 and 2002. Additionally, a disproportionate number of lesions were recorded in the frontal cortex and caudate nucleus compared to other brain regions. These lesion findings are consistent with other studies in the aging dog that suggest that the frontal lobes may be particularly vulnerable to age-related changes. PMID- 15795048 TI - Effects of scopolamine challenge on regional cerebral blood volume. A pharmacological model to validate the use of contrast enhanced magnetic resonance imaging to assess cerebral blood volume in a canine model of aging. AB - Cognitive impairment resulting from disruption of cholinergic function may occur through modulation of cerebrovascular volume (CBV). In the present study, dynamic susceptibility contrast magnetic resonance imaging (DSC-MRI) was used to examine cerebrovascular volume in young and old dogs during baseline and after administration of a cholinergic antagonist (scopolamine). In the first study, 24 animals (2-15 years of age) were given a baseline scan followed by a second scan after scopolamine administration (30 microg/kg). Gray matter rCBV was significantly higher than white matter rCBV during baseline and scopolamine administration. In the second study a subset of 7 dogs (4 young and 3 old) received scopolamine before anesthesia was induced for a second DSC-MRI scan. Consistent with the first study, gray matter rCBV was significantly higher than white matter rCBV. Scopolamine administered before anesthesia however, resulted in higher rCBV values compared to baseline in cerebral gray matter. Additionally, rCBVs were higher in young dogs at baseline in gray and white matter and marginally higher in gray matter when scopolamine was administered before anesthesia. These results indicate that in the dog, rCBV varies with brain compartment, decreases with age, and that DSC-MRI provides a measure of cerebrovascular function which may be related to age-dependent changes in cognition, brain structure, and neuropathology. PMID- 15795049 TI - Mitochondrial aging and dysfunction in Alzheimer's disease. AB - Disruptions in energy metabolism have been suggested to be a prominent feature, perhaps even a fundamental component, of Alzheimer's disease (AD). These abnormalities in cerebral metabolism precede the onset of neurological dysfunction as well as gross neuropathology of AD. These changes may stem from inhibition of mitochondrial enzymes including pyruvate dehydrogenase, cytochrome c oxidase, and alpha-ketoglutarate dehydrogenase. Several lines of evidence also suggest a role for oxidative stress in the neuropathology associated with the disease state. Because mitochondria are the major site of free radical production in cells, they are also a primary target for oxidative damage and subsequent dysfunction. This link between mitochondrial dysfunction and the pathophysiology of AD is supported by several lines of evidence. PMID- 15795050 TI - Further evidence for the cholinergic hypothesis of aging and dementia from the canine model of aging. AB - Memory decline in human aging and dementia is linked to dysfunction of the cholinergic system. Aging dogs demonstrate cognitive impairments and neuropathology that models human aging and dementia. This paper reviews recent evidence suggesting cholinergic involvement in canine cognitive aging based on studies with the anti-cholinergic drug, scopolamine, and a novel acetylcholinesterase inhibitor, phenserine. In particular, we examine: (1) the cognitive specificity of scopolamine's impairment in dogs, (2) the effect of age on scopolamine impairment and (3) the effect of phenserine on cognitive performance in dogs. Our findings indicate that working memory performance is disrupted by scopolamine at doses that do not disrupt non-cognitive behavior or long-term, semantic-like, memory, as indicated by performance of previously learned discriminations. This pattern of deficits is also seen in human and canine aging. We demonstrate that aged dogs are more sensitive to the impairing effects of scopolamine than young dogs, suggesting a decrease in cholinergic tone with increasing age. Dogs receiving phenserine demonstrate improved learning and memory compared to placebo controls. Our findings suggest that cholinergic decline could result in memory impairment, but that the memory impairment may be secondary to deficits in attention and/or encoding of new information. Together, these results suggest that the canine cholinergic system declines with age and that the aged dog is a unique model for screening therapeutics and for examining the relationship between amyloid pathology and cholinergic dysfunction in age dependent cognitive decline. PMID- 15795051 TI - Estrogen modulates cognitive and cholinergic processes in surgically menopausal monkeys. AB - Estrogen deficiency in postmenopausal women is associated with changes in physiological processes. The extent to which estrogen loss is associated with cognitive changes noted by postmenopausal women has been more difficult to determine for a variety of reasons. Primate models of menopause are now being used to determine the effects of estrogen loss and replacement on cognitive abilities and to investigate the neural mechanisms by which estrogen may influence cognitive function. The present report presents data from cognitive and neurobiological studies in surgically menopausal monkeys that have examined how estrogen loss and replacement may be affecting cognitive abilities and the cholinergic system; a neural system that is known to influence memory and attention function. These studies are indicating that visuospatial attention function is especially sensitive to estrogen states in young monkeys, but that multiple cognitive domains are sensitive to estrogen states in middle-aged monkeys. In addition, anatomical and functional imaging studies indicate that the primate cholinergic system is modulated by estrogen, and pharmacological studies demonstrate that estrogen uses cholinergic muscarinic receptors to influence visuospatial attention. These studies demonstrate that estrogen influences cognitive abilities in monkey models of menopause and the cholinergic system may be one of the mechanisms by which estrogen modulates cognitive function. Given the current unknowns and concerns regarding the use of hormone replacement therapy in postmenopausal women, continued studies in monkey models of menopause are especially needed to further elucidate the effects of estrogen on cognitive and neurobiological processes, with particular emphasis on studies in middle-aged monkeys, determining the optimal aspects of ERT regimens, and identifying the relationships between estrogen effects on cognitive and neurobiological function. PMID- 15795052 TI - Age-associated cognitive deficits in humans and dogs: a comparative neuropsychological approach. AB - We compared performance of younger and older human participants to that of younger and older dogs on tasks that evaluate object discrimination, egocentric spatial ability, object recognition, spatial memory, and cognitive flexibility. Our goal was to determine whether (i) tasks sensitive to advanced age in dogs are also age-sensitive in humans; (ii) the pattern of task difficulty observed in dogs mirrors that observed in humans; (iii) dogs and humans use similar strategies to solve equivalent tasks. Dogs performed more poorly than humans on reversal tasks that evaluate cognitive flexibility. We suggest that dogs, most notably older dogs, use different strategies than healthy humans when solving these tasks. Humans appear to test a priori hypotheses to solve the task at hand. As a consequence, expectations about the complexity of the task being tested can impair human performance. By contrast, dogs appear to rely more heavily on either simpler hypotheses, or associative trial and error learning, which probably accounts for their difficulty in learning non-matching tasks. Dogs also show perseverative responding, which hinders the acquisition of reversal tasks. PMID- 15795053 TI - Learning and memory as a function of age in Down syndrome: a study using animal based tasks. AB - Individuals with Down syndrome (DS) are at a high risk for developing Alzheimer disease (AD) after the age 40; however, low levels of intellectual functioning, coupled with impaired language ability, confound the detection of AD. Comparative neuropsychological tests developed in animal models of aging and cognition do not require intact language function and can be useful for detecting changes in cognition. Experimental paradigms used to detect age-dependent cognitive deficits in animal models were applied in the present study to measure cognitive function in a group of 20 adults with DS ranging in age from 22 to 58 years. Object discrimination, reversal learning, and spatial and object memory were administered using a modified Wisconsin General Testing Apparatus and reinforcement (penny rewards). When considering age as the only clinical variable to parallel the animal studies, age was significantly correlated with performance on object memory and marginally related to performance on reversal learning and spatial memory. However, when evaluating multiple clinical variables including age, a measure of intellectual ability (FSIQ), scores on the Dementia Questionnaire for Persons with Mental Retardation (DMR), and gender using regression analysis, scores on the DMR were the best predictors of errors of reversal learning, whereas FSIQ was the best predictor of performance on object memory. These results suggest that while age may be related to performance on learning and memory tasks, other clinical variables may be stronger predictors of performance in adults with DS. These changes may reflect prefrontal and medial temporal lobe dysfunction that is associated with the development of AD pathology in DS. PMID- 15795054 TI - Cognitive and behavioral assessment in dogs and pet food market applications. AB - A multi-disciplinary program was developed to assess the efficacy of antioxidant inclusion in a canine pet food on cognitive decline in aged beagles. A systematic approach to development of the food was used prior to beginning the cognitive studies. Comprehensive evaluation of antioxidant ingredients included assessments of commodities with naturally occurring antioxidants and synthetic antioxidants not commonly utilized, or at different concentrations than what was commonly utilized, in commercial pet foods. Studies were conducted to insure stability through processing, absorption from the gastrointestinal tract, safety, and tests for potential antioxidant biological benefit by ex vivo tests. Testing of the antioxidant-fortified food in aged beagles slowed the rate of cognitive decline in aged dogs. In addition, environmental enrichment also slowed the rate of cognitive decline. Importantly, the combination of dietary antioxidants and environmental enrichment was synergistic and resulted in the least amount of cognitive decline over the 30-month study period. Finally, a clinical study showed that antioxidant fortified food improved age-related behavioral changes in older pet dogs at in-home situations. PMID- 15795055 TI - Chronic antioxidant and mitochondrial cofactor administration improves discrimination learning in aged but not young dogs. AB - The present experiment was part of a 3-year longitudinal study examining the effects of age and antioxidant treatment on cognitive decline in beagles. Two size-concept tasks were administered following pretraining on a series of two choice (six subtests) and three-choice size discrimination tasks. Thirty-nine young and aged dogs were matched for age and cognitive ability then divided into four treatment groups. A combined antioxidant-mitochondrial cofactor treatment led to significantly improved performance in aged dogs on the first subtest of the two-choice size discrimination series. Treated aged dogs did not significantly differ from the young. Aged dogs on the antioxidant diet continued to perform better than aged controls on the second and third subtests, but these effects did not achieve significance. Young dogs performed significantly better than the aged dogs on the second and third subtests. The remaining two-choice tasks of the discrimination series were comparatively easy, leading to a floor effect. The antioxidant animals performed better on the three-choice size discrimination, but not on the two size-concept tasks. Antioxidants improved the performance of aged dogs on the initial learning tests, suggesting a selective improvement of factors related to the aging process and specific cognitive processes rather than general cognitive enhancement. PMID- 15795056 TI - Therapeutic agents for the treatment of cognitive dysfunction syndrome in senior dogs. AB - With increasing age, dogs develop a form of neurodegenerative disease which has many similarities to age related cognitive impairment and Alzheimer's disease in humans. A decline in learning and memory can be demonstrated in dogs beginning as young as 7 years of age using a variety of neuropsychological tests. However, clinical cases of cognitive dysfunction syndrome are seldom identified until the age of 11 years or older. This is likely due to the fact that the owners are relying on clinical observations such as house-soiling, sleep-wake cycles and disorientation, rather than tests of learning and memory. On the other hand, dogs that are trained to more exacting tasks such as guide dogs for the visually impaired, or bomb detection and agility trained dogs might be noticed to have a decline in performance at a much earlier age. Through the use of standardized neuropsychological testing protocols, a number of drugs, natural products and supplement formulations have been developed for use in dogs with cognitive dysfunction and, in some cases clinical trials have validated their efficacy. Furthermore, the testing of products currently licensed and in the pipeline for the treatment of cognitive decline and Alzheimer's in humans, may provide additional therapeutic agents for the treatment of senior dogs, as well as provide insight as to the potential for the efficacy of these compounds in humans. This review will examine those products that are now marketed along with some that might be considered for use in senior dogs with cognitive dysfunction as well as the research that has been used to validate the efficacy (or lack thereof) of these compounds. PMID- 15795057 TI - Effect of feeding patterns on performance of a visuospatial memory task in the beagle dog: a novel cognitive-based protocol for assessing satiety. AB - The assessment of appetite suppressing effects, or satiating effects, of drugs or other treatments is typically based on the measurement of food consumption and body weight. The present study describes a novel cognitive-based protocol for assessing satiety in the dog based on response latency and performance accuracy on a canine test of spatial working memory, the three-component delayed-non matching-to-position task (3cDNMP). We hypothesized that satiety, produced by providing food prior to testing, would reduce motivation to respond quickly and accurately on this food-reinforced task. Dogs were first over-trained on a variable-delay version of the 3cDNMP task. They were then pre-fed with either a single or a double portion of food prior to being tested on the same task. Pre feeding slowed response latency, but had no effect on performance accuracy. A more pronounced increase in response latency was observed in young dogs than in old dogs when offered double portions of food. These results suggest, first, that spatial working memory capability is independent of motivation; second, that satiety is age sensitive; and third, that a cognitive protocol can provide a reliable method for evaluating the satiating effects of various foods and other compounds in the dog. PMID- 15795058 TI - The canine model of human cognitive aging and dementia: pharmacological validity of the model for assessment of human cognitive-enhancing drugs. AB - For the past 15 years we have investigated the aged beagle dog as a model for human aging and dementia. We have shown that dogs develop cognitive deficits and neuropathology seen in human aging and dementia. These similarities increase the likelihood that the model will be able to accurately predict the efficacy of Alzheimer's disease (AD) treatments as well as detect therapeutics with limited or no efficacy. Better predictive validity of cognitive-enhancing therapeutics (CETs) could lead to enormous cost savings by reducing the number of failed human clinical trials and also may reduce the likelihood of negative outcomes such as those recently observed in the AN-1792 clinical trials. The current review assesses the pharmacological validity of the canine model of human aging and dementia. We tested the efficacy of (1) CP-118,954 and phenserine, two acetylcholinesterase inhibitors, (2) an ampakine, (3) selegiline hydrochloride, two drugs that have failed human AD trials, and (4) adrafinil, a putative CET. Our research demonstrates that dogs not only develop isomorphic changes in human cognition and brain pathology, but also accurately predict the efficacy of known AD treatments and the absence or limited efficacy of treatments that failed clinical trials. These findings collectively support the utilization of the dog model as a preclinical screen for identifying novel CETs for both age-associated memory disorder and dementia. PMID- 15795059 TI - Radioprotective effect of sesamol on gamma-radiation induced DNA damage, lipid peroxidation and antioxidants levels in cultured human lymphocytes. AB - Sesamol pretreated (1, 5 and 10 microg/ml) lymphocytes were exposed to different doses of gamma-radiation, i.e., 1, 2 and 4 Gray (Gy) and the cellular changes were estimated by using cytokinesis blocked micronucleus assay (MN), dicentric aberration (DC), thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Radiation significantly increased MN, DC frequencies, TBARS levels and decreased GSH and antioxidant enzyme levels in a dose dependent manner. The highest damage to lymphocytes was observed at 4 Gy irradiation. On the other hand, sesamol pretreatment significantly decreased MN, DC frequencies, TBARS levels and increased GSH levels and SOD, CAT and GPx activities in a concentration dependent manner. At 1 Gy irradiation all concentrations of sesamol (1, 5 and 10 microg/ml) significantly protects the lymphocytes from radiation damage. At 2 Gy irradiation 5 and 10 microg/ml of sesamol shows significant radioprotection. Since the highest damage was observed at 4 Gy irradiation both 1 and 5 microg/ml of sesamol pretreatment were not sufficient to protect the lymphocytes from radiation damage but 10 microg/ml of sesamol significantly (p<0.05) protects the lymphocytes from radiation effect. Thus, sesamol pretreatment gives significant protection to cultured human lymphocytes against gamma-radiation induced cellular damage. The possible mechanism involved in the radioprotective influence of sesamol is discussed. PMID- 15795060 TI - Blockade of haem oxygenase and nitric oxide synthetase causes cortical dysfunction in sheep exposed to carbon monoxide. AB - Twenty adult ewes underwent common surgery and following recovery were exposed to 1% carbon monoxide (CO) for 2 h. Ten of these sheep were randomly selected for treatment with haem oxygenase (HO) and nitric oxide synthetase (NOS) blockers. All sheep were killed 5 days later. The CO exposure was narcotic and EEG frequency was suppressed. The EEG recovery was rapid in the control sheep and both slow and incomplete in the treated sheep. This difference was statistically significant (P<0.05). For the first time in our CO studies in sheep, one showed multiple cortical infarcts. This sheep was blocked for HO and NOS function. No significant differences were seen in peri-ventricular white matter infarction distribution and frequency. We have previously shown brain protection against CO- and inert diluent-hypoxemia by way of an increase in brain blood flow (BBF) that maintains adequate brain O2 uptake, and by an increase in circulating red blood cells. From this study, we propose that the induction of neuronal and glial HO and NOS in sheep exposed to CO is protective, especially for the cortex. We intend to study this further by both selective and collective enzyme blockade and by measuring regional BBF changes. PMID- 15795061 TI - Maternal exposure of rats to nicotine via infusion during gestation produces neurobehavioral deficits and elevated expression of glial fibrillary acidic protein in the cerebellum and CA1 subfield in the offspring at puberty. AB - Maternal smoking during pregnancy is known to be a significant contributor to developmental neurological health problems in the offspring. In animal studies, nicotine treatment via injection during gestation has been shown to produce episodic hypoxia in the developing fetus. Nicotine delivery via mini osmotic pump, while avoiding effects due to hypoxia-ischemia, it also provides a steady level of nicotine in the plasma. In the present study timed-pregnant Sprague Dawley rats (300-350 g) were treated with nicotine (3.3 mg/kg, in bacteriostatic water via s.c. implantation of mini osmotic pump) from gestational days (GD) 4 20. Control animals were treated with bacteriostatic water via s.c. implantation of mini osmotic pump. Offspring on postnatal day (PND) 30 and 60, were evaluated for changes in the ligand binding for various types of nicotinic acetylcholine receptors and neuropathological alterations. Neurobehavioral evaluations for sensorimotor functions, beam-walk score, beam-walk time, incline plane and grip time response were carried out on PND 60 offspring. Beam-walk time and forepaw grip time showed significant impairments in both male and female offspring. Ligand binding densities for [3H]epibatidine, [3H]cytisine and [3H]alpha bungarotoxin did not show any significant changes in nicotinic acetylcholine receptors subtypes in the cortex at PND 30 and 60. Histopathological evaluation using cresyl violet staining showed significant decrease in surviving Purkinje neurons in the cerebellum and a decrease in surviving neurons in the CA1 subfield of hippocampus on PND 30 and 60. An increase in glial fibrillary acidic protein (GFAP) immuno-staining was observed in cerebellum white matter as well as granular cell layer of cerebellum and the CA1 subfield of hippocampus on PND 30 and 60 of both male and female offspring. These results indicate that maternal exposure to nicotine produces significant neurobehavioral deficits, a decrease in the surviving neurons and an increased expression of GFAP in cerebellum and CA1 subfield of hippocampus of the offspring on PND 30 and 60. The results show that although 60-day-old male and female rat offspring of mothers exposed to nicotine during gestation did not differ from control in body weight gain or nicotinic acetylcholine receptors ligand binding, they exhibited significant sensorimotor deficits that were consistent with the neuropathological alterations seen in the brain. These neurobehavioral and pathological deficits indicate that maternal nicotine exposure may produce long-term adverse health effects in the offspring. PMID- 15795062 TI - Tributyltin-induced effects on MAP kinases p38 and p44/42 in human natural killer cells. AB - NK cells form a class of lymphocytes that are able to kill tumor cells, virally infected cells and antibody-coated cells without prior sensitization. Tributyltin (TBT) is a toxic chemical that was used in large scale in wood preservation, marine antifouling paints, and slime control in paper mills. TBT has been detected in human foods such as dairy products, meat and fish and detectable levels have been found in human blood. The role of TBT in immunosuppression has been reported and its specific inhibition of the human NK-cell cytotoxic function has also been described. The current studies examined the role of mitogen activated protein kinases (MAPKs) p38 and p44/42 during TBT-induced inhibition of NK cytotoxic function. Continuous in vitro exposure to 300 nM TBT for 1h decreased the NK cell cytotoxic function with concomitant increases in phosphorylated forms of both p38 and p44/42 but not the total levels of either of these enzymes. Similar results were obtained with 500 nM TBT treatment. A 1 h exposure to 300 nM TBT followed by 24 h in TBT-free media resulted in a further decrease in the cytotoxic function with an accompanying increase in the phosphorylated forms of p38 and p44/42 MAPKs as compared to controls (approximately 50%). However, after 48 h in TBT-free media, the phosphorylation levels of both p44/42 and p38 MAPKs decreased by about 50 and 30%, respectively. NK cell cytotoxic function also continued to decrease with time in TBT-free media. A 24h exposure to 200 nM TBT decreased cytotoxic function of NK cells (>90%) but produced donor-dependent effects on the phosphorylated p38 and p44/42 MAPK levels. These data indicated that in vitro exposures to TBT induced changes in the phosphorylation (activation) states of p38 and p44/42 but not in their overall levels of expression in human NK cells. PMID- 15795063 TI - Benzoquinone activates the ERK/MAPK signaling pathway via ROS production in HL-60 cells. AB - Benzene (BZ) is a class I carcinogen and its oxidation to reactive intermediates is a prerequisite of hematoxicity and myelotoxicity. The generated metabolites include hydroquinone, which is further oxidized to the highly reactive 1,4 benzoquinone (BQ) in bone marrow. Therefore, we explored the mechanisms underlying BQ-induced HL-60 cell proliferation by studying the role of BQ-induced reactive oxygen species (ROS) in the activation of the ERK-MAPK signaling pathway. BQ treatment (0.01-30 microM) showed that doses below 10 microM did not significantly reduce viability. ROS production after 3 microM BQ treatment increased threefold; however, catalase addition reduced ROS generation to basal levels. FACS analysis showed that BQ induced a fivefold increase in the proportion of cells in S-phase. We also observed a high proportion of Bromodeoxyuridine (BrdU) stained cells, indicating a higher DNA synthesis rate. BQ also produced rapid and prolonged phosphorylation of ERK1/2 proteins. Simultaneous treatment with catalase or PD98059, a potent MEK protein inhibitor, reduced cell recruitment into the S-phase and also abolished the ERK1/2 protein phosphorylation induced by BQ, suggesting that MEK/ERK is an important pathway involved in BQ-induced ROS mediated proliferation. The prolonged activation of ERK1/2 contributes to explain the increased S-phase cell recruitment and to understand the leukemogenic processes associated with exposure to benzene metabolites. Thus, the possible mechanism by which BQ induce HL-60 cells to enter the cell cycle and proliferate is linked to ROS production and its growth promoting effects by specific activation of regulating genes known to be activated by redox mechanisms. PMID- 15795064 TI - Sex-specific effects of neonatal exposures to low levels of cadmium through maternal milk on development and immune functions of juvenile and adult rats. AB - Cadmium (Cd) is a major environmental contaminant. Although immunotoxic effects have been associated with Cd exposure, the inconsistency of experimental results underlines the need of an experimental approach more closely related to environmental conditions. We investigated the effects of exposing neonatal Sprague-Dawley rats to environmentally relevant doses of Cd through maternal milk. Dams received 10 parts per billion (ppb) or 5 parts per million (ppm) Cd chloride (CdCl2) in drinking water from parturition until the weaning of the pups. Half of the offspring was sampled at weaning time. The remaining juvenile rats received water without addition of Cd until adulthood. Cd accumulation in kidneys of juvenile rats fed from dams exposed to Cd indicated the transfer of the metal from mother to pups through maternal milk. This neonatal exposure resulted in decreased body, kidney and spleen weights of just weaned females but not of males. This effect was more pronounced in the less exposed females fed from dams exposed to 10 ppb Cd, which also displayed lower hepatic metallothionein-1 (MT-1) mRNA levels. The effect of Cd exposure on body and organ weights did not persist to adulthood. In contrast, we observed gender-specific effects of neonatal Cd exposure on the cytotoxic activity of splenic NK-cells of both juvenile and adult rats. Cd also strongly inhibited the proliferative response of Con A-stimulated thymocytes in both male and female adult rats 5 weeks after the cessation of Cd exposure. These immunotoxic effects were observed at doses much lower than those reported to produce similar effects when exposure occurred during adulthood. In conclusion, neonatal exposures to environmentally relevant levels of Cd through maternal milk represent a critical hazard liable to lead to both transitory and persistent immunotoxic effects. PMID- 15795065 TI - Stimulus control in animals: A tribute to the contributions of Donald S. Blough. PMID- 15795066 TI - Selective and divided attention in animals. AB - This article reviews some of the research on attentional processes in animals. In the traditional approach to selective attention, it is proposed that in addition to specific response attachments, animals also learn something about the dimension along which the stimuli fall (e.g., hue, brightness, or line orientation). More recently, there has been an attempt to find animal analogs to methodologies originally applied to research with humans. One line of research has been directed to the question of whether animals can locate a target among distracters faster if they are prepared for the presentation of the target (search image and priming). In the study of search image, the target is typically a food item and the cue consists of previous trials on which the same target is presented. In research on priming effects, the cue is typically different from the target but is a good predictor of its occurrence. The study of preattentive processes shows that perceptually, certain stimuli stand out from distracters better than others, depending not only on characteristics of the target relative to the distracters, but also on relations among the distracters. Research on divided attention is examined with the goal of determining whether an animal can process two elements of a compound sample with the same efficiency as one. Taken together, the reviewed research indicates that animals are capable of centrally (not just peripherally) attending to selective aspects of a stimulus display. PMID- 15795067 TI - Object discrimination by pigeons: effects of object color and shape. AB - Can nonhuman animals attend to visual stimuli as whole, coherent objects? We investigated this question by adapting for use with pigeons a task in which human participants must report whether two visual attributes belong to the same object (one-object trial) or to different objects (two-object trial). We trained pigeons to discriminate a pair of differently colored shapes that had two targets either on a single object or on two different objects. Each target equally often appeared on the one-object and two-object stimuli; therefore, a specific target location could not serve as a discriminative cue. The pigeons learned to report whether the two target dots were located on a single object or on two different objects; follow-up tests demonstrated that this ability was not entirely based on memorization of the dot patterns and locations. Additional tests disclosed predominate stimulus control by the color, but not by the shape of the two objects. These findings suggest that human psychophysical methods are readily applicable to the study of object discrimination by nonhuman animals. PMID- 15795068 TI - How do pigeons represent numbers? Studies of number scale bisection. AB - Pigeons' bisection of two number scales, 1-16 and 2-32, was examined in two experiments. In Experiment 1, pigeons learned to choose different comparison keys following 1 or 16 key pecks and 2 or 32 key pecks and then were tested with intermediate numbers of key pecks on nonreinforced probe trials. Psychophysical curves that plotted choice of the large number key against number of pecks showed superposition of the two number scales and indifference between the two keys at the geometric means of each scale. The same procedures were used in Experiment 2, but pigeons were trained to bisect each scale at its arithmetic mean. Two asymmetries appeared in curves that plotted discrimination performance against number: near the midpoints of the scales, accuracy was higher for high numbers than for low numbers, but this relationship reversed at the ends of the scales, with low numbers discriminated more accurately than high numbers. An associative model that assumed stimulus generalization between numbers was used to predict the findings of these two experiments. The model showed a good qualitative fit to the obtained data when it was assumed that number was represented on a log scale and generalization was constant at all numbers but not when it was assumed that number was represented on a linear scale and generalization was scalar. PMID- 15795069 TI - A Turing test of a timing theory. AB - A quantitative theory of timing or conditioning can be evaluated with a Turing test in which the behavioral results of an experiment can be compared with the predicted results from the theory. An example is described based upon an experiment in which 12 rats were trained on three fixed-interval schedules of reinforcement, and a simulation of the predicted results from a packet theory of timing. An objective classification rule was used to determine whether a sample from the data or a sample from the theory was more similar to another sample from the theory. With an ideal theory, the expected probability of a correct classification would be 0.5. The observed probability of a correct classification was 0.6, which was slightly, but reliably, greater than 0.5. A Turing test provides a graded metric for the evaluation of a quantitative theory. PMID- 15795070 TI - Better living by not completing: a wonderful peculiarity of pigeon vision? AB - Whereas many non-human species have been demonstrated to visually complete partly occluded figures, pigeons have been repeatedly failed to do so. We asked whether this failure reflected the pigeons' lack of perceptual process for completion or their decision among completed and non-completed figures. Four pigeons searched for a red lozenge target having one of its four contours punched in a rectangular edge out among three intact lozenges. All of these four stimuli had a white square next to them. After obtaining consistent search performances, the pigeons were tested with the punched target in a variety of locations relative to the white square, including right at the edges. Humans tested in the same task needed longer times before detecting the target when the square was placed right at the punched edge, suggesting automatic completion in humans. In contrast, the pigeons showed no similar difficulty. This result has two important suggestions: first, pigeons fail to complete partially occluded objects at the perceptual level, and second, this lack of completion is sometimes advantageous for them. PMID- 15795071 TI - Pigeons do not perceptually complete partly occluded photos of food: an ecological approach to the "pigeon problem". AB - Humans routinely complete partly occluded objects to recognize the whole objects. However, a number of studies using geometrical figures and even conspecific images have shown that pigeons fail to do so. In the present study, we tested whether pigeons complete partially occluded objects in a situation simulating a natural feeding context. In Experiment 1, we trained pigeons to peck at any photograph of food and not to peck at any containing a non-food object. At test, we presented both photos of food partly occluded by pigeon's feather and photos simply truncated at the same part. We predicted that if the pigeons perceptually completed the occluded portion, then they would discriminate the photos of occluded food better than the truncated photos. The result was that the pigeons pecked at the truncated photos earlier than the occluded photos. Placing the occluder next to all of the stimuli in Experiment 2 or substituting indented lozenge for the feather in Experiment 3 did not affect the results. Thus, even in a simulated ecologically significant situation, pigeons continued to not show evidence of perceptual completion. PMID- 15795072 TI - Avian detection and identification of perceptual organization in random noise. AB - Recent research has suggested that pigeons may have difficulty globally integrating visual information in hierarchically arranged stimuli. To isolate and understand the mechanisms responsible for processing emergent perceptual structure, three pigeons were tested in a two alternative choice task that required the global integration of organized local information. They were reinforced for localizing, on randomized distractor backgrounds of black and white square elements, different types of structured targets (e.g., stripes, squares, checkerboards) arranged from these same elements. These hierarchical stimuli were tested at four different levels of spatial granularity (i.e., different element sizes). Experiment 1 found rapid acquisition for the vertical and horizontal stripes or square targets and somewhat slower learning with the checkerboard pattern. Experiment 2 demonstrated successful transfer to a novel target types (alternating bars and "diagonal" stripes). In both experiments, displays with the greatest spatial granularity (smallest elements and most repetitive structure) monotonically supported the best discrimination. These results indicate pigeons can perceive and discriminate emergent visual structure under the right circumstances and suggest they do so with a generalized rule for detecting patterns of non-random perceptual structure. PMID- 15795073 TI - Canine distemper virus associated proliferation of canine footpad keratinocytes in vitro. AB - Infection of canine footpads with canine distemper virus (CDV) can result in so called hard pad disease characterized by footpad epidermal proliferation and hyperkeratosis. Cultured canine footpad keratinocytes (CFK) were inoculated with a virulent canine distemper virus strain (A75/17-CDV) to study the effects of CDV infection on keratinocyte proliferation. Infection was analyzed by immunohistochemistry and in situ hybridization for CDV nucleoprotein (N-protein) antigen and mRNA. CDV caused a persistent, non-cytocidal infection with spread from single cells to infection of the confluent cell layer 7 days post infection (p.i.). Absolute cell numbers were significantly higher in infected cultures compared to control cultures from day 4 until day 6 p.i. Infected cultures contained significantly more total DNA on day 5 p.i. compared to controls. Immunohistochemical investigation of proliferation markers Ki67 and BrdU demonstrated a nearly two-fold increase in numbers of positive cells on day 5 p.i. compared to controls. These findings demonstrate that canine distemper virus infection of canine footpad keratinocytes in vitro was associated with proliferation. PMID- 15795074 TI - Characterisation of three equine influenza A H3N8 viruses from Germany (2000 and 2002): evidence for frozen evolution. AB - Reported here are the results of antigenic and genetic characterisation of equine influenza strains causing local outbreaks reported to the Equine Diagnostic Centre in Berlin, Germany. In 2000, equine influenza virus was detected in a nasal swab from a non-vaccinated horse using a rapid diagnostic kit, but was not successfully isolated. Partial direct sequencing of the haemagglutinin (HA1) gene, indicated that the virus was a European lineage H3N8 subtype strain representative of strains isolated in several European countries during 2000. In 2002, two equine influenza viruses were isolated from nasal swabs both taken from unvaccinated horses with acute respiratory symptoms housed at the same stables. Antigenic characterisation using a panel of ferret antisera suggested that these isolates also belonged to the European lineage of H3N8 viruses. Analysis of deduced HA1 amino acid sequences confirmed that the HA1 of both isolates were identical and belonged to the European lineage. However, from phylogenetic analysis, both strains appeared to be more closely related to viruses isolated between 1989 and 1995 than to viruses isolated more recently in Europe. These results suggested that viruses with fewer changes than those on the main evolutionary lineage may continue to circulate. The importance of expanding current equine influenza surveillance efforts is emphasised. PMID- 15795075 TI - Transmission of caprine herpesvirus 2 in domestic goats. AB - Caprine herpesvirus 2 (CpHV-2) is a recently recognized gammaherpesvirus that is endemic in domestic goats and has been observed to cause clinical malignant catarrhal fever (MCF) in certain species of deer. In this study, transmission of CpHV-2 in goats was examined. A total of 30 kids born to a CpHV-2 positive goat herd were selected and divided into two groups: group 1 (n=16) remained in the positive herd; group 2 (n=14) was separated from the herd at 1 week of age after obtaining colostrum. Peripheral blood samples from each kid were examined regularly by competitive ELISA for MCF viral antibody and by PCR for CpHV-2 DNA. Fifteen out of 16 goats (94%) that remained with the positive herd seroconverted and became PCR-positive for CpHV-2 by 10 months of age. In contrast, all kids (100%) that were separated from the positive herd at 1 week of age remained negative until termination of the experiment at 1 year of age. Additional transmission experiments revealed that all CpHV-2-free adult goats were susceptible to CpHV-2 or ovine herpesvirus 2 (OvHV-2) infection. The data indicate that the transmission pattern of CpHV-2 in goats is similar to the pattern of OvHV-2 in sheep and that CpHV-2-free goats can be established by early separation of kids from positive herds, which has significant implications for MCF control programs. PMID- 15795076 TI - New insights into the genetic diversity of European porcine reproductive and respiratory syndrome virus (PRRSV). AB - The complete ORF5 sequences of 66 porcine reproductive and respiratory syndrome (PRRS) field virus strains (1991-2001) and three European modified live vaccine strains were determined, as well as ORFs 6 and 7 of 19 selected strains. The variability of the deduced ORF5 amino acid sequences was analysed using statistical process control (SPC), allowing for the objective assessment of variable and conserved regions. Four variable and four conserved regions as well as five hypervariable amino acid positions were defined. The effects of genetic variability on possible structural and functional properties were discussed with emphasis on immunogenic features. Phylogenetic analysis and pairwise comparison of the nucleotide sequences revealed that the genetic distances between the strains has greatly increased over time. The data do not support an evolutionary influence of the geographical location or the time of sample collection, nor of PRRSV vaccination on strain development. In contrast to other authors who tended to concentrate on the samples from either a common geographic origin or a short sampling period, we could not confirm geographically separate PRRSV clusters nor did we find evidence of positive selective pressure as measured by the ratio of synonymous to non-synonymous substitutions in ORF5, 6 or 7. Immunological implications and vaccination strategies are discussed. PMID- 15795077 TI - Diagnostic tests for small ruminant lentiviruses. AB - Maedi visna virus and caprine arthritis encephalitis virus are closely related retroviruses that cause chronic inflammatory disease in small ruminants. The infections are characterised by insidious onset and slow progression. Diagnosis of infection is usually by serological testing. A variety of assays are available for this purpose, though the relative sensitivity and specificity of these assays has not been compared systematically. Here we review recent developments in laboratory diagnostic methods and their use in field diagnosis. The results suggest that a combination of ELISA and PCR might afford optimal detection of SRLV infection. PMID- 15795078 TI - Biochemical analysis, cpn60 and 16S rDNA sequence data indicate that Streptococcus suis serotypes 32 and 34, isolated from pigs, are Streptococcus orisratti. AB - Streptococcus suis serotypes have traditionally been identified by morphology, biochemical profiling and serotyping. Analysis of the sequences of 16S rRNA and cpn60 genes of the 35 characterized serotypes of S. suis led to the observation that two serotypes 32 and 34, are significantly distinct from other S. suis serotypes and may represent a distinct species. Here we present DNA sequence data and biochemical profiles which indicate that S. suis serotypes 32 and 34, isolated from pigs, are clustered with Streptococcus orisratti, a Voges-Proskauer negative, alpha-haemolytic, aesculin-hydrolytic, Lancefield group A streptococcus isolated from the teeth of rats. PMID- 15795080 TI - An attenuated Salmonella Enteritidis strain derivative of the main genotype circulating in Uruguay is an effective vaccine for chickens. AB - We have recently reported that Salmonella enterica serovar Enteritidis (S. Enteritidis) strains circulating in Uruguay, are unevenly distributed among different genetic subtypes, with a predominant genotype that is a common contaminant of poultry-derived food and that accounts for the vast majority of human cases of food-borne disease. Herein, we describe the construction of a genetically-defined aroC derivative (LVR02) of a local strain of S. Enteritidis belonging to the major genetic type. We demonstrated the attenuation and the immunogenicity of that strain in a mouse model, and evaluated it as a vaccine for commercial layer chickens. LVR02 proved to be stable, attenuated, innocuous, immunogenic and to induce protective immunity against a S. Enteritidis challenge when used for oral vaccination. A single oral dose of LVR02 administered to newly hatched chickens induced protection against oral challenge with the parental virulent strain, preventing systemic and persistent intestinal infection and significantly reducing the shedding of the challenge strain in birds' feces. A second vaccine dose at 15 days post-hatching boosted the immunogenicity of the vaccine, and strengthened the protection achieved with a single dose. This strain may represent the basis of a live vaccine to be included in national control programs to reduce circulation of this pathogen in the country. PMID- 15795079 TI - Seroprevalence of anaplasmosis among cattle in Switzerland in 1998 and 2003: no evidence of an emerging disease. AB - Anaplasma marginale infection in Europe has been limited to the Mediterranean and eastern countries, to Austria and to very sporadic cases in Switzerland. There are no reports of its occurrence in the countries north of Switzerland. A severe outbreak of anaplasmosis in August 2002 in a cattle farm in the canton Grisons, Switzerland, north of the Alps, with more than 300 cattle that had to be culled, came unexpected and gave reason to hypothesize presence of an increased yet undetected prevalence of A. marginale in Switzerland. Randomly selected bovine serum samples collected in 1998 and 2003 were tested using a competitive inhibitory ELISA (cELISA) to test the hypothesis. Our validation of the diagnostic sensitivity and specificity of this test, done in the outbreak herd, yielded 99.2 and 83.3%, respectively, probably underestimating the true specificity. The true seroprevalence of anaplasmosis in Swiss cattle determined by cELISA was likely to be zero with upper 95% confidence limits of 2.49% in the canton Grisons and 1.17% in the rest of Switzerland, respectively, in 1998. For 2003, these estimates were even lower. There was no significant difference in apparent prevalences between 1998 and 2003. In search of a possible reservoir, three chamoises out of 46 free ranging wild ruminants from the Swiss National Park, Grisons, tested positive in the cELISA. This reaction is in accordance with A. marginale or a cross reacting agent such as Anaplasma ovis. From our results we conclude that the hypothesis of an increased prevalence of anaplasmosis in cattle in Switzerland must be rejected. PMID- 15795081 TI - Key role of Chlamydophila psittaci on Belgian turkey farms in association with other respiratory pathogens. AB - Two hundred turkey sera from eight Belgian and two French farms were tested for the presence of antibodies against avian pneumovirus (APV), Ornithobacterium rhinotracheale (ORT), Mycoplasma gallisepticum, Mycoplasma meleagridis and Chlamydophila psittaci. At slaughter, C. psittaci, APV and ORT antibodies were detected in 94, 34 and 6.5% of the turkeys, respectively. No antibodies against M. gallisepticum or M. meleagridis were present. Additionally, turkeys on three Belgian farms were examined from production onset until slaughter using both serology and antigen or gene detection. All farms experienced two C. psittaci infection waves, at 3-6 and 8-12 weeks of age. Each first infection wave was closely followed by an ORT infection starting at the age of 6-8 weeks, which was still detectable when the second C. psittaci infection waves started. Animals on farm A were not vaccinated against APV leading to an APV subtype B outbreak accompanying the first C. psittaci infection wave. Despite subtype A APV vaccination on farms B and C, the second C. psittaci infection waves were accompanied (farm B) or followed (farm C) by a subtype B APV infection. On all farms respiratory signs always appeared together with a proven C. psittaci, APV and/or ORT infection. This study suggests an association between C. psittaci, APV and ORT, and indicates the multi-factorial aetiology of respiratory infections in commercial turkeys. All three pathogens should be considered when developing prevention strategies for respiratory disease. PMID- 15795082 TI - Variable and strain dependent colonisation of chickens by Escherichia coli O157. AB - The prevalence of enterohaemorrhagic Escherichia coli (EHEC) O157 in poultry is considered minimal compared with other species, especially ruminants. However, deliberate inoculation studies have shown that poultry are readily and persistently infected by this organism but that the mechanism of colonisation is independent of intimin, a recognised factor in host-EHEC interactions in mammalian species, and may be dependent upon flagella. Few strains of EHEC O157 have been tested in poultry and here 1-day-old and 6-week-old chicks were inoculated with seven non-toxigenic E. coli O157 strains in separate experiments. Persistence was measured semi-quantitatively by bacteriological assessment of E. coli O157 cultured from cloacal swabs (shedding score). In the 1-day-old chick model that was monitored for 43 days, all seven strains established well after inoculation. In the 6-week-old chicken model, one strain established and gave consistently high shedding for the duration of the experiment (156 days). Whereas of the remaining six strains, two persisted for 113 days, two persisted for 43 days, one persisted for 22 days and one strain was never detected. PMID- 15795084 TI - Possible associations between Salmonella persistence in poultry houses and resistance to commonly used disinfectants and a putative role of mar. AB - A putative link between Salmonella persistence in the agricultural sector and resistance to disinfectants has been sparsely investigated. Therefore, minimum inhibitory concentration (MIC) tests against five disinfectants commonly used in poultry premises (formaldehyde, glutaraldehyde/benzalkonium chloride compound, oxidising compound, tar oil phenol, iodophor) were performed on 286 Salmonella isolates, including 256 from Danish broiler houses, altogether representing nine serotypes. Six of these isolates were used for adaptation and de-adaptation studies involving the five disinfectants. Amongst 60 of these isolates selected for growth studies in cyclohexane (possibly associated with up-regulated efflux), only one isolate grew. From this isolate and the six isolates used in the adaptation and de-adaptation studies, mutants highly resistant to triclosan (a disinfectant linked with mar-type resistance) were selected. In addition, adaptation and de-adaptation studies with triclosan were performed. For the 286 isolates, the small variations in MICs could not be associated with Salmonella persistence in Danish broiler houses or previous use of relevant disinfectants. Adaptation and de-adaptation did not alter MICs to the five farm disinfectants. Compared to the parent isolates, MICs for the triclosan adapted and de-adapted isolates and the triclosan mutants were significantly increased to triclosan, but not to the five disinfectants. Moreover, most of the triclosan adapted and de adapted isolates grew in cyclohexane. Thus, there was no correlation between triclosan and cyclohexane resistance on one hand and resistance to the five disinfectants on the other, suggesting that triclosan resistance is not linked with resistance to these disinfectants. PMID- 15795083 TI - Comparison of fecal culture and Danish Mix-ELISA for determination of Salmonella enterica subsp. enterica prevalence in growing swine. AB - In the USA, control of food-borne salmonellosis associated with meat consumption has been predominantly focused at slaughter and processing. It is expected that standards at slaughter and processing will become more stringent, creating pressure to reduce prevalence of Salmonella-positive food animals through on-farm interventions. The aim of this study was to compare traditional fecal culture and the Danish Mix-ELISA (DME) for determination of Salmonella prevalence pre-harvest in swine. In Trial 1, five cohorts of individually identified pigs were longitudinally sampled during the growing period to compare the kinetics of prevalence as estimated by fecal culture and the DME. In Trial 2, the correlation between fecal prevalence and seroprevalence was estimated pre-marketing in 49 groups of pigs. In Trial 1, fecal prevalence and seroprevalence showed similar kinetics, with a tendency of a higher OD% cut-off to more closely approximate fecal prevalence. In Trial 2, correlations between fecal culture and the DME were 0.40, 0.36, 0.43, and 0.43 (p<0.001) for OD% cut-offs > or =10, 20, 30, and 40, respectively. Based on these results, a higher OD% cut-off would be recommended if more approximate estimation of fecal prevalence is desired and longitudinal sampling would be suggested for evaluating the impact of on-farm interventions for Salmonella reduction whether utilizing fecal culture or the DME. Further evaluation of the impact of Salmonella serovar present on farms on seroprevalence and the relationship of on-farm seroprevalence with food safety risk are needed prior to utilizing the DME for pre-harvest Salmonella diagnostics in the US swine herd. PMID- 15795086 TI - Amplification of 16S rRNA genes of Anaplasma species in China for phylogenetic analysis. AB - In this study, a phylogenetic tree was inferred through comparing five 16S rRNA gene sequences of four isolates of Anaplasma ovis and one of Anaplasma marginale in China with all nineteen 16S rRNA gene sequences deposited in GenBank (12 A. marginale, 3 A. ovis and 4 Anaplasma centrale derived from America, Uruguay, South Africa, Zimbabwe, Australia, Isreal and Japan). The analysis showed that all A. ovis isolated in China were separated into an A. ovis cluster, while the A. marginale in China was separated into an A. marginale cluster (see Fig. 1). This analysis demonstrated that there are at least two different Anaplasma species widespread among ruminants in North China. PMID- 15795085 TI - A real-time PCR assay to detect the Panton Valentine Leukocidin toxin in staphylococci: screening Staphylococcus schleiferi subspecies coagulans strains from companion animals. AB - Recent reports suggest that methicillin-resistant strains of Staphylococcus schleiferi subspecies coagulans are now commonly isolated from dogs. Given the association of a potentially mobile SCCmec type IV element with lysogenic phage encoded Panton Valentine Leukocidin (PVL) toxin genes in community-acquired methicillin-resistant Staphylococcus aureus strains we hypothesized that methicillin-resistant S. schleiferi ssp. coagulans strains may also encode PVL toxin genes. Forty S. schleiferi ssp. coagulans strains isolated from companion animals were studied. Susceptibility to oxacillin was determined by broth microdilution and all isolates were screened by PCR for the presence of the mecA gene. SCCmec typing was performed on 14 isolates. A real-time PCR assay was developed for the detection of the PVL genes using a SmartCycler. Pulsed-field gel electrophoresis (PFGE) was performed to determine whether S. schleiferi ssp. coagulans strains were homogeneous. Twenty-eight of the 40 isolates (70%) were resistant to oxacillin and 26/28 possessed the mecA gene by PCR. SCCmec IV was identified in seven strains; the other seven isolates were not typable by this technique. All 40 strains were negative for the PVL toxin gene. PFGE showed a heterogeneous population and 13 different profiles were determined. In conclusion, this study showed that PVL toxin genes were not detected in a heterogeneous population of methicillin-resistant S. schleiferi ssp. coagulans strains isolated from companion animals. PMID- 15795087 TI - The use of multilocus enzyme electrophoresis to characterise intestinal spirochaetes (Brachyspira spp.) colonising hens in commercial flocks. AB - Multilocus enzyme electrophoresis (MLEE) was used to identify, examine genetic relationships and look at disease associations of a collection of 53 intestinal spirochaete isolates previously recovered from the faeces of adult hens on 14 farms in Qld, Australia. The MLEE results were compared with those previously obtained using species-specific PCR amplifications. The isolates were divided into five Brachyspira species groups by MLEE: Brachyspira murdochii (n=17), B. intermedia (n=15), B. pilosicoli (n=14), B. innocens (n=2) and "B. pulli" (n=1). Three new MLEE groups each containing single isolates also were identified. The results of the PCR assay for B. pilosicoli were concordant with the MLEE results, but the 23S rDNA-based PCR for B. intermedia had failed to detect 8 of the 15 isolates. The B. innocens/B. murdochii nox-based PCR had correctly identified all the isolates of B. murdochii, but did not identify either of the two B. innocens isolates. Using MLEE, isolates from two farms (14%) were identified as B. murdochii, whilst the pathogenic species B. intermedia and B. pilosicoli were present in hens from eight (57%) and five (36%) farms, respectively, and were identified together in four (29%) farms. All seven of the farms with production problems or wet litter were colonised with B. intermedia and/or B. pilosicoli. Six farms had multiple spirochaete isolates available for examination. Two broiler breeder farms both had five isolates of B. pilosicoli that shared the same MLEE electrophoretic type (ET), whilst one laying hen farm had three isolates of B. intermedia that all belonged to the same ET. Hence on each of these farms a predominant strain of a pathogenic species was present. On the other farms isolates of the same species were more diverse and belonged to different ETs. These results show that the epidemiology of intestinal spirochaetal infections in broiler breeder and laying hen flocks can vary considerably between farms, although the reasons for these differences were not established. PMID- 15795088 TI - Cholesterol as target of Fe-NTA-induced lipid peroxidation in rat tissues. AB - Intraperitoneal injection of the iron-chelate, ferric-nitrilotriacetate (Fe-NTA), induces renal proximal tubular damage associated with oxidative damage in vivo. A sub-lethal dose of Fe-NTA (15 mg Fe/kg body weight) was administered IP to rats; animals were sacrificed and liver, kidney and plasma were collected 1-4 h after injection. In response to the Fe-NTA administration, there were significant time dependent reductions of the levels of total lipids, cholesterol and total unsaturated fatty acids, and a rise in the concentrations of conjugated dienes, 7 ketocholesterol and fatty acids hydroperoxides, showing a pattern inversely correlated in plasma, kidney and liver. Cholesterol level decreased significantly from 1 h after injection in the kidney and 3-4 h in the plasma and liver of treated rats. This is the first report on cholesterol reduction and accumulated 7 ketocholesterol in the tissues of rats treated with Fe-NTA as a consequence of lipid peroxidation. PMID- 15795089 TI - Limited modulation of the transport activity of the human multidrug resistance proteins MRP1, MRP2 and MRP3 by nicotine glucuronide metabolites. AB - The human ATP-binding cassette proteins MRP1 (ABCC1), MRP2 (ABCC2) and MRP3 (ABCC3) are active transporters of antineoplastic drugs as well as conjugated metabolites and other organic anions. In addition to being substrates, many glucuronide, glutathione and sulfate conjugates can also inhibit the transport activities of these MRP-related proteins, sometimes in a glutathione (GSH) dependent manner. Nicotine is the major addictive component of cigarette smoke. Three glucuronide metabolites of this compound have been identified in vivo: nicotine-N-glucuronide, cotinine-N-glucuronide and trans-hydroxycotinine-O glucuronide. In this study, we first chemically synthesized trans-hydroxycotinine O-glucuronide and then tested the ability of this compound, nicotine-N glucuronide and cotinine-N-glucuronide to modulate the vesicular transport of several organic anions by MRP1, MRP2 and MRP3. We observed that none of the three metabolites at concentrations up to 100muM significantly affected organic anion transport by MRP1 or MRP2, either in the absence or presence of GSH. MRP3 mediated transport of 17beta-estradiol 17-(beta-d-glucuronide) and methotrexate were partially inhibited by trans-hydroxycotinine-O-glucuronide (300 microM) (by 70% and 50%, respectively), whereas nicotine-N-glucuronide and cotinine-N glucuronide had no effect. We conclude that the physiological functions of MRP1, MRP2 and MRP3 are not likely to be substantially affected by nicotine glucuronide metabolites at concentrations achievable in human serum. PMID- 15795090 TI - Effects of genistein on the expression of hepatic genes in two rat strains (Sprague-Dawley and Wistar). AB - Information about the effects of phytoestrogens in the liver, the main organ of metabolism, is limited. In this study we have examined the dose dependent effects of the phytoestrogen genistein (GEN) on gene expression in rat liver in comparison to the potent estrogen ethinylestradiol (EE). Special attention was paid to strain specific differences between Wistar- (WIS) and Sprague-Dawley rats (SD). EE as well as GEN stimulated the gene expression of insulin like growth factor binding protein 1 (IGFBP-1), similarly in both strains. The mRNA expression of Vitamin D dependent calcium-binding protein (CaBP9k) was stimulated by EE in both strains but more pronounced in WIS rats. Interestingly GEN stimulates CaBP9k gene expression only in SD rats. Gene expression of apolipoprotein AI (ApoAI) was not affected in both strains neither by EE nor by GEN. These results are indicative of strain specific differences in the response of liver genes to GEN or EE. PMID- 15795091 TI - The cumulative activity of benzo(a)pyrene on systemic immune responses with mite allergen extract after intranasal instillation and ex vivo response to ovalbumin in mice. AB - The cumulative activity of benzo(a)pyrene (BP) as one of the polycyclic aromatic hydrocarbons which is present in diesel exhaust particles on systemic immune responses was studied in mice after intranasal instillation and in vitro exposure to allergens. The main purpose was to elucidate whether BP has an adjuvant effect in this responses or not. Male Balb/cA mice were immunized with mite allergen extract (MAE) either alone or in combination with BP. Controls were given BP, or buffer alone. The animals were immunized four times. One week after the last immunization, anti-MAE IgE antibody in serum and IL-4, IL-10, IL-12p70 and IFN gamma secreted by spleen cells upon rechallenge with the same allergen, was measured by ELISA. An increased response to the antigen was observed in animals receiving MAE together with BP, compared with animals receiving MAE alone. In another attempt, bone marrow-derived dendritic cells (DCs) are obtained from naive male Balb/c mice and were exposed to the varying doses of ovalbumin (OA) and BP. There were four groups of study: control, BP, OA and OA plus BP. An elevated response was observed in animals receiving OA together with BP, compared with animals receiving OA alone. In conclusion, this work shows that BP has an cumulative activity for specific IgE and Th1/Th2 cytokines production after intranasal instillation and cytokine secretion by DCs after in vitro exposure, and this phenomenon is dose-dependent. Thus, it may be another evidence on immunomodulatory effect of dendritic cells. PMID- 15795092 TI - Transcriptional induction of CYP3A4 by o,p'-DDT in HepG2 cells. AB - Technical grade DDT, a commonly used pesticide, contains 20% of the o,p'-DDT isomer. It alters the activity of hepatic mixed function oxidase and induces expression of cytochrome P450 such as 3A subfamilies. In the present study, we analyzed the effects of o,p'-DDT on CYP3A4 gene expression in HepG2 cells. Treatment with o,p'-DDT dose-dependently increased levels of CYP3A4 mRNA and the immunoreactive protein, by 13- and 3.8-fold, respectively, relative to untreated cultures. Treatment with actinomycin D blocked the o,p'-DDT-induced increase in CYP3A4 mRNA levels. These findings demonstrate that this insecticide is able to induce CYP3A4 transcription and may play an important role in the modulation of endogenous hormones and xenobiotic metabolism. PMID- 15795094 TI - Cadmium-induced nephropathy in the development of high blood pressure. AB - In recognition of a central role of the kidney in long-term blood pressure control, we undertook an in-depth analysis of the relationship between blood pressure and kidney damage caused by environmental exposure to the common pollutants cadmium and lead. The subjects were 200 healthy Thais, 16 and 60 years of age (100 female non-smokers, 53 male non-smokers, and 47 male smokers). None of these subjects had been exposed to Cd or Pb in the workplace and their urinary Cd concentrations ranged from 0.4 to 37 nM, whereas their urinary Pb concentrations ranged from 0.1 to 30 nM. The prevalence of high blood pressure was 2%, 8% and 19%, respectively in subjects with low, average and high Cd-burden (linear trend chi2=6.4, P=0.01). Multiple regression analysis revealed a significant positive association between Cd-burden and blood pressure in male non smokers (adjusted beta=0.31, P=0.02) and an inverse association between blood pressure and urinary Pb excretion rate in male smokers (adjusted beta=-0.38, P=0.005). Associations between Cd-burden and nephropathies were evidenced by increases in urinary excretion of beta2-microglobulin (P=0.02) and N-acetyl-beta d-glucosaminidase (P=0.005) in subjects with high Cd-burden, compared with the subjects with average Cd-burden. In addition, an association between Cd-related nephropathy and high blood pressure was evidenced by a 20% increase in the prevalence of high blood pressure in people with NAG-uria (linear trend chi2=4.3, P=0.04). Our present study provides first evidence for a possible link between renal tubular damage and dysfunction caused by environmental Cd exposure and increased risk of high blood pressure. PMID- 15795093 TI - Myristicin-induced neurotoxicity in human neuroblastoma SK-N-SH cells. AB - Myristicin, 1-allyl-3,4-methylenedioxy-5-methoxybenzene, is a naturally occurring alkenylbenzene compound found in the nutmeg. The present study was conducted to assess the cytotoxic and apoptotic effects of myristicin on the human neuroblastoma SK-N-SH cells. We found that a dose-dependent reduction in cell viability occurs at myristicin concentration > or =0.5 mM in SK-N-SH cells. Apoptotic cell death was confirmed using DNA fragmentation, terminal deoxyribonucelotidyl transferase-mediated dUTP nick end labeling and by 4,6 diamidino-2-phenylindole staining. Microscopy was used to observe apoptotic cell morphology. Western blotting was used to investigate the protein expression of known apoptotic mediators including cytochrome c, caspase-3, and PARP. The apoptosis triggered by myristicin was accompanied by an accumulation of cytochrome c and by the activation of caspase-3. The results obtained suggest that myristicin induces cytotoxicity in human neuroblastoma SK-N-SH cells by an apoptotic mechanism. This myristicin-induced apoptosis provides further insight of the molecular mechanisms of myristicin toxicity. PMID- 15795096 TI - Time and concentration dependent accumulation of [3H]-deltamethrin in Xenopus laevis oocytes. AB - A primary target of pyrethroid insecticides are the voltage-sensitive sodium channels (VSSCs). Expression of VSSCs in oocytes from Xenopus laevis is an experimental model used to study the effects of pyrethroids. A common assumption when utilizing this model is that media concentration is an accurate substitute for tissue dose. This assumption may not hold true for lipophilic chemicals. [3H] deltamethrin (DLT) was used to test the hypothesis that media concentration is a good surrogate for tissue concentration. Accumulation of DLT (0.001-10 microM) in non-transfected oocytes exposed for 20 min was determined using liquid scintillation counting. The time course (1.0-180 min) of tissue accumulation of DLT (approximately 1.0 microM (0.50 ppm) in media) was also determined. Results demonstrate that tissue dose increases as a function of time with media concentration underestimating tissue dose at long incubation times (approximately 2.0-fold at 180 min) and overestimating tissue dose short incubation times (approximately 8.6-fold at 5 min). Tissue dose increases as a function of media concentration, with overestimation of tissue dose ranging from 1.5-fold at 0.0005 ppm to 4.1-fold at 5.0 ppm. These data suggest that media concentration does not accurately predict tissue dose at all times for a broad range of deltamethrin concentrations in X. laevis oocytes. PMID- 15795095 TI - Metallothionein overexpression and resistance to toxic stress. AB - Metallothionein (MT) protects against the harmful effects of a wide spectrum of stress factors. The most studied of these factors is cadmium, whose toxicity is reduced on sequestration by MT. However, there is poorer consensus in the literature about protection afforded by MT against stressors other than cadmium. In this study, a CHO-K1 cell line continuously overexpressing MT (MToex) was developed in order to evaluate the relative protection afforded by MT against different toxic agents. Cadmium was used as a positive control and, as expected, the MToex cells were more than 13-fold more resistant to the effects of cadmium chloride than were wild-type (WT) cells using the MTT (3-[4,5-dimethylthiazol-2 yl]-2,5-diphenyl tetrazolium bromide) assay (IC50 values of 10 and 132 microM for WT and MToex cells, respectively). In contrast, overexpression of MT afforded no protection against mercuric chloride, staurosporine and hydrogen peroxide (IC50 values of about 50, 11 and 925 microM, respectively). Cd and Hg uptake by MToex and WT cells exposed to 1-10 microM of metal chloride was similar and yet a significant amount of these metals was associated with the cytosol MT fraction in the MToex cells but not in the WT cells. From this study it can be concluded that while MT overexpression protects against Cd toxicity, it has no influence on Hg, staurosporine or hydrogen peroxide toxicity and it is proposed that this reflects mechanistic differences of toxicity or depletion of labile intracellular zinc by the presence of excess binding ligand in the form of MT. PMID- 15795097 TI - The 2004 French Medical Academy award to professor Georges Mathe. PMID- 15795098 TI - Modulation of P-glycoprotein function and reversal of multidrug resistance by (-) epigallocatechin gallate in human cancer cells. AB - Multidrug resistance (MDR) is a major obstacle in the chemotherapeutic treatment of many human cancers. In this study, the reversal of P-glycoprotein (P-gp) mediated multidrug resistance by (-)-epigallocatechin gallate (EGCG) and its molecular mechanism were investigated. A three-dimensional model of carboxyl terminal nucleotide binding domain (NBD2) from P-gp was built by homology modeling. The structural model of the complex indicates that EGCG was tightly bound to the ATP-binding site of NBD2. EGCG modulated the function of P-gp and increased the intracellular accumulation of chemotherapeutic agent doxorubicin (DOX) in drug-resistant KB-A1 cells. When KB-A1 cells were exposed to 10 microg/ml DOX combined with 10, 30, 50 microM EGCG for 4 h, the intracellular concentrations of DOX were increased 1.5, 1.9, 2.3 times, respectively compared with DOX alone treatment. In vitro EGCG potentiated the cytotoxicity of DOX to drug-resistant KB-A1 cells. In KB-A1 cell xenograft model, EGCG could also enhance the efficacy of DOX and increased the DOX concentration in the resistant tumors. Thus, these results suggest that EGCG modulated the function of P-gp and reversed P-gp mediated multidrug resistance in human cancer cells. PMID- 15795100 TI - Mechanistic explanation for the unique pharmacologic properties of receptor partial agonists. AB - Simulation data provided in this paper revealed that when a receptor full agonist and a competitive or noncompetitive antagonist for the same type of receptor are covalently linked together in some proper ways, the resulting bifunctional compound would have a reduced efficacy, and their dose-response curve patterns would look exactly like the curve patterns for many known partial agonists. It is also shown that all known pharmacologic characteristics for receptor partial agonists matched precisely the projected properties of the agonist-antagonist bifunctional compounds. The novel mechanistic explanation developed in this paper not only reveals the structural requirements for receptor partial agonists, but it also negates the long-held mechanistic explanation that the reduced efficacy of receptor partial agonists is the result of partial activation of each individual receptor molecule being occupied by the partial agonist because of its "low intrinsic activity". PMID- 15795099 TI - Activation of the coagulation system in cancerogenesis and metastasation. AB - The activation of the coagulation system in cancer patients is a well-known phenomenon responsible for recurrent clinical problems. A number of fascinating molecular mechanisms have been recognized showing that the tumor not only activates the coagulation system, but vice versa, activated coagulation proteins are able to induce molecular effects in tumor cells. The molecular basis is the expression of defined membrane receptors by tumor cells that are activated, for example, by thrombin. As the liberation of thrombin from prothrombin is one of the key events in coagulation, it's impact upon biological processes, such as cancerogenesis and metastasation, seems to be a regular pathophysiological consequence. These perceptions are not only interesting for the comprehension of cancerogenesis, metastasation, and clinical phenomena, but they also have a high impact upon modern strategies of tumor therapy. Especially, the development of clinically useful coagulation inhibitors, such as modern low molecular weight heparins or melagatran, created the possibility of therapies that combine cell biological approaches with apoptosis-inducing principals such as chemotherapy. Several clinical studies that demonstrate the implication of these strategies have already been published recently. In this article the cell biological basics for these approaches are reviewed. PMID- 15795101 TI - Mechanism of multidrug resistance in relation to passive membrane permeation. AB - Passive uptake of drugs into cells is described in terms of the following steps: (1) massive immediate binding of the drugs to the outer leaflet of the plasma membrane resulting in practical equilibrium between extremely high drug concentrations at the cell surface compared to the drug concentration in the medium. (2) Due to their amphipathic nature, anticancer drugs are practically excluded from the lipid core of the membrane. They cross the lipid core by distinct flip-flop events that occur in the case of doxorubicin and daunorubicin after an average period of 0.7 and 0.15 min, respectively. (3) The drug reaching the inner leaflet of the plasma membrane is in practical equilibrium with the drug present in the cytoplasm. (4) Almost all the amounts of anticancer drugs present in the cells are bound by molecular sinks, such as DNA or cytoskeleton elements. The resistance afforded to multidrug resistant (MDR) cells by extrusion pumps, such as P-glycoprotein, is negatively correlated with the affinity of the drugs to the membranes and with their flip-flop rates across membranes. Binding rates of the drugs to membranes and intracellular sinks have no effect on drug concentration in the cytoplasm once equilibrium is reached between the passive uptake of drugs and their active extrusion. PMID- 15795102 TI - Repressions of MMP-9 expression and NF-kappa B localization are involved in inhibition of lung carcinoma 95-D cell invasion by (-)-epigallocatechin-3 gallate. AB - Epigallocatechin-3-gallate (EGCG) repressed the invasion of lung carcinoma 95-D cells in invasion assay. RT-PCR analysis illuminated that 40 microM EGCG down regulated the expression of MMP-9 by 45.7% and the result of Western blot analysis provided further evidence. NF-kappa B localized in the nucleus of the 95 D cells was diminished in a dose-dependent manner in EGCG-treated cells as shown by Western blot. Intracellular oxidants were more abundant in invasive cells than in invasion-suppressed cells fed with EGCG for 18 h. Thus, the inhibition of tumor invasion by EGCG was shown to be attributed to decreases of the expression of MMP-9 and NF-kappa B, which may result from decrease of intracellular oxidants. PMID- 15795104 TI - Contrast-enhanced dynamic MR imaging parameters and histological types of invasive ductal carcinoma of breast. AB - The diagnosis of breast cancer has progressed owing to the improvement of both MRI equipments and MR techniques. CE-MRI is expected to be more useful for diagnosis of the existence and characteristics of breast cancer than mammography and ultrasound. Forty-three breast cancer patients, who underwent breast MRI examination before surgery and had histologically confirmed invasive ductal carcinomas, were included in this study. In 19, seven and 17 of them, carcinomas were histologically classified as papillotubular, solid-tubular, and scirrhous type, respectively. In dynamic MRI studies, the MR parameters were calculated from the signal intensity determined before and at five time-points after a bolus of 0.1 mmol Gd-DTPA/kg in the chosen region of interest (ROI). The differential analysis of MR parameters with reference to histopathology was performed by one way analysis of variance (ANOVA) in dynamic studies. Four of 15 parameters analyzed, ENH1-2, ENH1, MSLP, and SAT3, were significantly different between papillotubular and scirrhous carcinoma. The dynamic curve of papillotubular carcinoma reached a maximum in the very early phase and then decreased slowly (linear pattern), while that of scirrhous carcinoma peaked in the slightly later phase and then decreased (parabolic pattern). In the differential analysis by Kruskal-Wallis test, although no significant differences were found in any morphological parameters among three histological types, spicula formation and rim enhancement tended to appear comparatively frequently in scirrhous and solid tubular carcinoma. CE-MRI might reveal the differences of growth characteristics among different histological types of breast carcinomas and provide a diagnostic method useful for estimation of the prognosis of breast cancer. PMID- 15795103 TI - Physiological, pharmacological and clinical features of the multidrug resistance protein 2. AB - Multidrug resistance protein 2 (MRP2, ABCC2) is a drug efflux pump belonging to the ATP-binding cassette (ABC) transporter superfamily. MRP2 is present predominantly at the biliary pole of hepatocytes and is also expressed in the kidney and intestine. It plays a major role in hepato-biliary elimination of many structurally diverse xenobiotics, including organic anions and drug conjugates, and therefore most likely contributes to pharmacokinetic parameters of these compounds. MRP2 also handles endogenous molecules such as bilirubin, and its overexpression has been shown to confer a multidrug resistance phenotype to tumoral cells. MRP2 expression can be regulated by endogenous substances such as inflammatory cytokines and biliary acids. The MRP2 levels and activity can also be affected by a large panel of xenobiotics, including chemopreventive agents and ligands of the pregnane X receptor, which may be a potential source of drug-drug interactions and drug adverse effects. MRP2 appears therefore as one of the major drug efflux pumps of the organism, whose functional and regulatory features are important to consider, notably for drug disposition. PMID- 15795105 TI - Effect of 5-azacytidine and procainamide on CD3-zeta chain expression in Jurkat T cells. AB - It has been observed that decrease of DNA methyltransferase 1 (DNMT1) activity is associated with low content of the CD3-zeta (zeta) chain in T cell receptor (TCR)/CD3 complex of T cells in systemic lupus erythematosus (SLE) patients. The CD3-zeta chain plays a pivotal role in intracellular signal transmission between TCR/CD3 complex and nuclei. The compounds 5'-azacytidine (AZC) and procainamide (PCA) belong to inhibitors of DNMT1, whose low activity correlates with increase in transcription of various genes. Using the reverse-transcription and real-time quantitative PCR (RQ-PCR) analysis, we indicated that AZC and PCA did not profoundly affect on CD3-zeta chain transcription in Jurkat T leukemia cells clone E6-1. However, the flowcytometric analysis revealed that AZC and PCA decreased intracellular contents of CD3-zeta chain in these cells in dose dependent manner. Our results suggest that decrease of DNMT1 activity may alter intracellular signal transmission without effect on transcription level of CD3 zeta chain. PMID- 15795106 TI - Epidemiological and clinical features of 139 patients with tuberculosis at a teaching hospital in Italy (Pisa, 1996-2000). AB - In order to describe epidemiologic and clinical features of patients with tuberculosis (TB) identified recently in the hospital of Pisa (Tuscany, Italy), a retrospective study of all cases of TB notified to the Local Public Health Service during January 1996-December 2000 was performed. The diagnosis of TB was made following the criteria of the WHO. A total of 139 patients affected by TB were identified. Diagnosis was microbiologically proved in 81 patients. Mean age was 53.8+/-20.5 S.D. yrs. Thirty-five (25.2%) patients were extra European community citizens (mostly from Africa). The incidence of TB (N/100.000) was 8.4 in 1996 and 6.8 in 2000. Sixty-eight point three per cent of patients had pulmonary TB, 24.5% extrapulmonary and 7.2% mixed TB. The rate of extrapulmonary TB was 15.9% and 39.2% in the 1996-98 and in the 1999-2000 periods, respectively (p = 0.002). Extrapulmonary TB was more frequent in extra European community citizens (42.8%) than in Italian ones (18.3%), p = 0.003. Seven patients were presenting also advanced HIV infection. Microscopic examination for acid fast bacilli in sputum or bronchial secretion resulted negative in 17.4% of proved pulmonary TB (positive culture for Mycobacterium tuberculosis). The chest x-rays showed pleural effusion in 19 patients. Pulmonary cavitation was documented in 15 patients with negative chest x-rays. Fever was not present in 42.4% of the patients at the moment of diagnosis. Three point eight percent of the isolated strains of M. tuberculosis were in vitro multidrug-resistant. The data presented showed an important rate of TB in Pisa. We have yet to understand if the decreased rate observed in 2000 represents a new trend as reported in other North American and European countries. The rate of extrapulmonary TB shows a trend to increase accordingly to recent literature. The isolation rate of multidrug resistant strains of M. tuberculosis in Pisa seems to be similar to the rates reported in other areas of Europe. PMID- 15795107 TI - Incorporating protein transduction domains (PTD) within recombinant 'fusion' transcription factors. A novel strategy for directing stem cell differentiation? AB - Application of embryonic and adult stem cells in regenerative medicine will require efficient protocols for directing stem cell differentiation into well defined lineages. The use of exogenous cytokines, growth factors, or extracellular matrix substratum, will obviously require extended durations of in vitro culture. With autologous adult stem cells, this could delay transplantation to the patient, as well as alter the immunogenicity of the cultured autologous cells. Genetic modulation to direct stem cell differentiation would obviate prolonged durations of in vitro culture; but there are overwhelming safety concerns with regards to the application of recombinant DNA technology in human clinical therapy. A novel alternative would be to incorporate protein transduction domains (PTD) into recombinant transcription factors that play important roles in somatic differentiation. Such protein-engineered transcription factors will then have the ability to translocate across the cell membrane and be internalized within the cytosol, thereby acting as paracrine signaling molecules. Upon internalization, the recombinant transcription factors would only have a limited active half-life, so that their effects are only transient. However, this could provide sufficient stimulus for initiating stem cell differentiation into a required lineage. PMID- 15795108 TI - Herpes simplex virus resistance to acyclovir in routine virological laboratory practice. PMID- 15795109 TI - Cogent confabulation. AB - A new model of vertebrate cognition is introduced: maximization of cogency p(alphabetagammadelta|epsilon). This model is shown to be a direct generalization of Aristotelian logic, and to be rigorously related to a calculable quantity. A key aspect of this model is that in Aristotelian logic information environments it functions logically. However, in non-Aristotelian environments, instead of finding the conclusion with the highest probability of being true (a popular past model of cognition); this model instead functions in the manner of the 'duck test;' by finding that conclusion which is most supportive of the truth of the assumed facts. PMID- 15795110 TI - Intelligent initialization of resource allocating RBF networks. AB - In any neural network system, proper parameter initialization reduces training time and effort, and generally leads to compact modeling of the process under examination, i.e. less complex network structures and better generalization. However, in cases of multi-dimensional data, parameter initialization is both difficult and time consuming. In the proposed scheme a novel, multi-dimensional, unsupervised clustering method is used to properly initialize neural network architectures, focusing on resource allocating networks (RAN); both the hidden and output layer parameters are determined by the output of the clustering process, without the need for any user interference. The main contribution of this work is that the proposed approach leads to network structures that are compact, efficient and achieve best classification results, without the need for manual selection of suitable initial network parameters. The efficiency of the proposed method has been tested on several classes of publicly available data, such as iris, Wisconsin and ionosphere data. PMID- 15795111 TI - Chaotic dynamics of a behavior-based miniature mobile robot: effects of environment and control structure. AB - To study the regularity and complexity of autonomous behavior, the flow of sensory information obtained in autonomous mobile robots under various conditions was analyzed as a complex system. Sensory information time series Xn was collected from a miniature mobile robot during free navigation, and plotted on the return map, the graph of X(n+tau) vs. Xn. The plot exhibited a characteristic trajectory, representing the regularity of the time series. Correlation integral and Lyapunov exponent analysis also showed properties of deterministic chaos; the presence of fractal dimension and positive Lyapunov exponent. Analysis of sensory information obtained in the robot with three different neural controllers revealed that the autonomous robot behaves in such a way that the flow of sensory information is governed by a deterministic rule, and this pattern is unique to each controller. Furthermore, the analysis in various environments exhibited that transitions from one trajectory to another on the return map occur during the course of autonomous behavior. The fractal and Lyapunov dimensions calculated in various conditions indicate that these dimension could be utilized to quantify the complexity of autonomous behavior and the relative difficulty of tasks. Analyses at different evolutionary stage revealed that behavioral performance correlates with fractal dimension. These studies using a miniature mobile robot that allowed to idealize the experimental conditions demonstrated firmly that the complex analysis could be utilized in evaluation and optimization of autonomous systems and the behavior. PMID- 15795113 TI - Analysis of global exponential stability and periodic solutions of neural networks with time-varying delays. AB - In this paper, a general class of recurrent neural networks with time-varying delays is studied. Some novel and sufficient conditions are given to guarantee the global exponential stability of the equilibrium point and the existence of periodic solutions for such delayed neural networks. Comparing with some previous literature, in which the time-varying delays were assumed to be differentiable and their derivatives were simultaneously required to be not greater than 1, the restrictions on the time-varying delays are removed. Therefore, our results obtained here improve and extend some previously related results. Finally, two numerical examples are provided to illustrate our theorems. PMID- 15795114 TI - On the global output convergence of a class of recurrent neural networks with time-varying inputs. AB - This paper studies the global output convergence of a class of recurrent neural networks with globally Lipschitz continuous and monotone nondecreasing activation functions and locally Lipschitz continuous time-varying inputs. We establish two sufficient conditions for global output convergence of this class of neural networks. Symmetry in the connection weight matrix is not required in the present results which extend the existing ones. PMID- 15795112 TI - Adaptive algorithms for first principal eigenvector computation. AB - The paper presents a unified framework to derive and analyze 10 different adaptive algorithms, some well-known, to compute the first principal eigenvector of the correlation matrix of a random vector sequence. Since adaptive principal eigenvector algorithms have originated from a diverse set of disciplines, including ad hoc methods, it is necessary to examine them in a unified framework. In a common framework consisting of five steps, we analyze the derivation, convergence, and rate results for many well-known algorithms as well as two new adaptive algorithms. In the process, we offer fresh perspectives on the known algorithms, and derive new results for others. The common framework also allows us to comparatively study the 10 algorithms. Finally, we show experimental results to support our analyses. PMID- 15795115 TI - NEUROM: a ROM based RNS digital neuron. AB - In this work, a fast digital device is defined, which is customized to implement an artificial neuron. Its high computational speed is obtained by mapping data from floating point to integer residue representation, and by computing neuron functions through residue arithmetic operations, with the use of table look-up techniques. Specifically, the logic design of a residue neuron is described and complexity figures of area occupancy and time consumption of the proposed device are derived. The approach was applied to the logic design of a residue neuron with 12 inputs and with a Residue Number System defined in such a way as to attain an accuracy better than or equal to the accuracy of a 20-bit floating point system. The proposed design (NEUROM) exploits the RNS carry independence property to speed up computations, in addition it is very suitable for using look up tables. The response time of our device is about 8 x T(ACC), where T(ACC) is the ROM access time. With a value of T(ACC) close to the 10 ns allowed by the current ROM technology, the proposed neuron responds within 80 ns, NEUROM is therefore the neuron device proposed in the literature which allows for maximum throughput. Moreover, when a pipeline mode of operation is adopted, the pipeline delay can assume a value as low as about 14 ns. In the case study considered, the total amount of ROM is about 5.55 Mbits. Thus, using current technology, it is possible to integrate several residue neurons into a single VLSI chip, thereby enhancing chip throughput. The paper also discusses how this amount of memory could be reduced, at the expense of the response time. PMID- 15795116 TI - TAO-robust backpropagation learning algorithm. AB - In several fields, as industrial modelling, multilayer feedforward neural networks are often used as universal function approximations. These supervised neural networks are commonly trained by a traditional backpropagation learning format, which minimises the mean squared error (mse) of the training data. However, in the presence of corrupted data (outliers) this training scheme may produce wrong models. We combine the benefits of the non-linear regression model tau-estimates [introduced by Tabatabai, M. A. Argyros, I. K. Robust Estimation and testing for general nonlinear regression models. Applied Mathematics and Computation. 58 (1993) 85-101] with the backpropagation algorithm to produce the TAO-robust learning algorithm, in order to deal with the problems of modelling with outliers. The cost function of this approach has a bounded influence function given by the weighted average of two psi functions, one corresponding to a very robust estimate and the other to a highly efficient estimate. The advantages of the proposed algorithm are studied with an example. PMID- 15795117 TI - Experimentally optimal nu in support vector regression for different noise models and parameter settings. PMID- 15795118 TI - Regulatory roles of sex hormones in cutaneous biology and immunology. AB - Recent studies have revealed that sex hormones manifest a variety of biological and immunological effects in the skin. Pregnancy, menstruation and the menopause modulate the natural course of psoriasis, indicating a female hormone-induced regulation of skin inflammation. Estrogen in vitro down-regulates the production of the neutrophil, type 1 T cell and macrophage-attracting chemokines, CXCL8, CXCL10, CCL5, by keratinocytes, and suppresses IL-12 production and antigen presenting capacity while enhancing anti-inflammatory IL-10 production by dendritic cells. These data indicate that estrogen may attenuate inflammation in psoriatic lesions. Estrogen, alone or together with progesterone, prevents or reverses skin atrophy, dryness and wrinkles associated with chronological or photo-aging. Estrogen and progesterone stimulate proliferation of keratinocytes while estrogen suppresses apoptosis and thus prevents epidermal atrophy. Estrogen also enhances collagen synthesis, and estrogen and progesterone suppress collagenolysis by reducing matrix metalloproteinase activity in fibroblasts, thereby maintaining skin thickness. Estrogen maintains skin moisture by increasing acid mucopolysaccharide or hyaluronic acid levels in the dermis. Progesterone increases sebum secretion. Estrogen accelerates cutaneous wound healing stimulating NGF production in macrophages, GM-CSF production in keratinocytes and bFGF and TGF-beta1 production in fibroblasts, leading to the enhancement of wound re-innervation, re-epithelialization and granulation tissue formation. In contrast, androgens prolong inflammation, reduce deposition of extracellular matrix in wounds, and reduce the rate of wound healing. Estrogen enhances VEGF production in macrophages, an effect that is antagonized by androgens and which may be related to the development of granuloma pyogenicum during pregnancy. These regulatory effects of sex steroids may be manipulated as therapeutic or prophylactic measures in psoriasis, aging, chronic wounds or granuloma pyogenicum. PMID- 15795120 TI - Analysis of T cell receptor (TCR) BV-gene clonotypes in NC/Nga mice developing dermatitis resembling human atopic dermatitis. AB - BACKGROUND: Our previous study showed that T cells in skin lesions of human atopic dermatitis (AD) had oligoclonal accumulation, indicating the involvement of antigen-specific immune reactions at those sites. Recently, NC/Nga mice, which develop skin lesions similar to AD, have been proposed as a model for that disease. OBJECTIVE: To clarify whether NC/Nga mice are suitable as a model for human AD from the viewpoint of their antigen-specific immune responses. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and single strand conformation polymorphism (SSCP) analyses were conducted to detect TCR BV genes of clonally expanded T cells derived from NC/Nga mice at an early phase of the AD like dermatitis, at a late phase of the dermatitis, and with no AD-like dermatitis. RESULTS: (1) T cells with TCR BV 7, 10 and 17 reside in the skin of NC/Nga mice without the AD-like dermatitis. (2) T cells with these BV genes contain oligoclonal accumulations, however, expanded T cell clonotypes are also detected in the spleen and exist constantly during the course of the AD-like dermatitis. (3) Development of the AD-like dermatitis is associated with additional oligoclonal expansion/accumulation of T cells with TCR BV 2, 4 and 6 genes. (4) Progression of the AD-like dermatitis is associated with further oligoclonal expansion/accumulation of T cells with the TCR BV 14 gene. (5) Some of the expanded TCR clonotypes are common between the individual mice and between early and late phases. CONCLUSIONS: Taking these data together with the previous human AD studies, NC/Nga mice seem to be an appropriate model for human AD. PMID- 15795119 TI - Immunohistochemical characterization of cutaneous drug eruptions by STI571. AB - BACKGROUND: STI571, a selective BCR-ABL tyrosine kinase inhibitor, is a promising new drug for chronic myelogenous leukemia (CML). However, the drug has been reported to be associated with adverse cutaneous drug eruptions with high frequency. OBJECTIVE: In this study, the characteristics of the cutaneous drug eruptions by STI571 were investigated. METHODS: The clinical records of 10 patients diagnosed with drug eruption by STI571 were reviewed. We obtained 10 skin biopsy specimens from patients with drug eruption by STI571, 6 from the antibiotics-induced drug eruption group, and 5 from normal skin (control). Immunohistochemical analysis was performed to detect CD4, CD8, CD56, IL-18, IL 1beta and ICAM-1 expression in the cutaneous drug eruption. RESULTS: Seven out of 10 patients had maculopapular exanthema, 2/10 erythema multiforme, 1/10 urticaria. We analyzed the composition of T-lymphocyte subsets from the infiltrates at the STI571-induced drug eruption site in eight patients. Unlike other drug eruptions, the increase in the CD8 expression was statistically significant, especially in the dermoepidermal junction and the upper dermis (P < 0.01). The enhanced expression of IL-18 and IL-1beta was observed as well. In contrast, ICAM-1 was either weakly positive or negative. CONCLUSION: Drug eruption caused by STI571 was mostly expressed as a maculopapular exanthema. The histopathological findings were similar in drug eruption by antibiotics or STI571. Unlike the drug eruptions caused by antibiotics, where the expression of CD4 was dominant, CD8 was dominant in drug eruptions by STI571. The expression of IL-18 and IL-1beta was increased in both groups. This elevation of IL-18 and IL 1beta may assist in understanding the pathogenesis of cutaneous drug eruption. PMID- 15795122 TI - Human epidermal basal cell responses to ultraviolet-B differ according to their location in the undulating epidermis. AB - BACKGROUND: Exposure of skin to excessive ultraviolet-B (UVB) radiation causes epidermal hyperproliferation that leads to epidermal hyperplasia, however, it is not yet clear exactly how these responses progress. OBJECTIVES: We attempted to clarify the response patterns involved with epidermal hyperproliferation following UVB radiation. METHODS: UVB was irradiated at 2 minimal erythema doses (MED) to human back skin and epidermal morphologic changes were evaluated using in vivo confocal laser microscopy. Skin biopsy specimens were collected from exposed and from non-exposed regions, and were subjected to histochemical and immunohistochemical analysis. RESULTS: The in vivo confocal laser microscopic analysis showed that UVB-induced epidermal hyperplasia was prominent at the epidermal rete ridges. Further, 3 days after UVB exposure, numerous Ki67-positive epidermal cells were observed in the epidermal rete ridges, but not in the epidermis at the top of the dermal papilla. These results suggest that cells highly responsive to UVB exist in the epidermal rete ridges and that their hyperproliferation leads to elongation of the epidermal rete ridges. In contrast, the number of keratin 10-positive basal cells, known as transitional cells, was increased throughout the epidermis, suggesting that an upward migration of keratinocytes from the epidermal basal layer occurred regardless of their location. However, diffusion of melanin to the suprabasal layers was markedly observed in epidermal regions above the dermal papillae, suggesting the occurrence of strong upper cell movement at this position. CONCLUSION: Based on our results, we conclude that differences in keratinocyte responses to UVB radiation exist in cells located in the undulating epidermal basal layer. PMID- 15795121 TI - HLA DR B1*04, *07-restricted epitopes on Keratin 17 for autoreactive T cells in psoriasis. AB - BACKGROUND: Psoriasis is a T cell-mediated inflammatory skin disease. Recent evidence suggests that activated CD4+ helper T lymphocytes of the Th1 phenotype play an important role in the pathogenesis of the disease. For psoriatic autoreactive T cells, Keratin 17 is a major target antigen and an epitope containing ALEEAN sequence has been described, but other psoriasis-related epitopes are still unknown. OBJECTIVE: To identify the HLA DR B1*04, *07 restricted T cell epitopes on Keratin 17. METHODS: HLA DRB1*04, *07-restricted T cell epitope regions on Keratin 17 were predicted based on related software and internet servers. Keratin 17 gene was amplified from psoriatic epidermis and the proteins of the predicted epitope regions were expressed, identified and purified. T cells from psoriatic patients reacted in cultivation with peptide major histocompatibility complex (p-MHC) compound, then the level of cell proliferation and the concentration of interferon-gamma in culture supernatant were detected. After the psoriasis-related epitope regions were narrowed down, the epitopes on them were predicted further. These epitopes were then expressed and validated by T cell response in vitro. RESULTS: Four epitopes--S1 (118-132), S2 (169-183), S4 (323-337) and S4 (348-362) can stimulate the proliferation and interferon-gamma production of psoriatic T cells more effectively than other epitopes (p<0.01) and react weakly with the T cells from healthy volunteers (p>0.05). CONCLUSION: Epitopes S1 (118-132), S2 (169-183), S4 (323-337) and S4 (348-362) are immunodominant DR B1-restricted T cell epitopes for psoriasis. Among them, S1 (118-132) contains the ALEEAN sequence while the others with different amino acid sequence have not been reported before. Further studies based on these peptides would provide a more complete understanding of the immunological basis of psoriasis. PMID- 15795123 TI - Nadifloxacin, an antiacne quinolone antimicrobial, inhibits the production of proinflammatory cytokines by human peripheral blood mononuclear cells and normal human keratinocytes. AB - BACKGROUND: Acne vulgaris is a chronic inflammatory disease involving colonization of Propionibacterium acnes (P. acnes), activation of neutrophils and lymphocytes. Circumstantial evidence suggests that antigen-independent and dependent immune responses against P. acnes are involved in the pathogenesis of inflammatory acne. Epidermal keratinocytes are also suggested to be involved in initiation and progression of cutaneous inflammation. Nadifloxacin, a fluorinated quinolone, has potent antimicrobial activities against Gram-negative and positive microbes and is used to treat multiple inflamed acne lesions. However, its effect on immune conferring cells such as mononuclear cells and keratinocytes has not been examined. OBJECTIVE: To evaluate the possible involvement of potential anti-inflammatory activity of nadifloxacin in its therapeutic effect on inflammatory acne, we examined the effects of nadifloxacin, in comparison with other antibiotics used to treat acne vulgaris, on cytokine production by human peripheral blood mononuclear cells (PBMC) and keratinocytes. METHODS: Cytokine production by PBMC was determined after treatment with heat-killed P. acnes in the presence or absence of antimicrobials using a real-time PCR and ELISA. Cultured human epidermal keratinocytes were stimulated by IFN-gamma plus IL-1beta and the effects of antimicrobials were examined by using ELISA. RESULTS: Nadifloxacin as well as macrolide antibiotics and clindamycin inhibited IL-12 and IFN-gamma production by PBMC stimulated by heat-killed P. acnes. The drug also inhibited the IL-1alpha, Il-6, IL-8 and GM-CMS production by keratinocytes treated with IFN-gamma plus IL-1beta. CONCLUSIONS: Inhibitory effects of nadifloxacin to activate T cells and keratinocytes may be involved at least in part in the mechanism of its therapeutic effect against inflammatory acne. PMID- 15795124 TI - IL-4 and cellular adhesion molecule (CAM) pathway are involved in cicatricial pemphigoid scarring process. PMID- 15795125 TI - Identification of a recurrent mutation in keratin 17 in a Japanese family with pachyonychia congenita type 2. PMID- 15795126 TI - Senescent human keratinocytes suppress colony formation of HeLa cells. PMID- 15795127 TI - Preferential downmodulation of certain chemokines by fexofenadine in human keratinocytes. PMID- 15795129 TI - Neurobiology of decision-making: quo vadis? AB - Decision-making is an important aspect of daily life. Moreover, dysfunctions of decision-making play a critical role in a number of neuropsychiatric disorders. Several important research groups have contributed a diverse set of approaches to study decision-making and its underlying neurobiology. Insights from these studies may provide important directions for future research in this area. PMID- 15795130 TI - Making sense of candidates: partisanship, ideology, and issues as guides to judgment. AB - Citizens employ a range of heuristic devices in reaching judgments and making evaluations regarding political candidates and issues. This paper examines the circumstances that lead to the use of multiple and potentially competing heuristics in making complex decisions. In our analysis, subjects systematically demonstrate evidence of confusion regarding particular combinations of partisan, ideological, and issue-based heuristics--a difficulty in disentangling partisan, ideological, and issue-based criteria in the evaluation of candidates. We argue that the political environment alters both the accessibility of particular heuristic devices, as well as the strength of association in memory among alternative heuristic devices. Hence the ability of citizens to select among heuristics is conditioned on the larger environment of incoming information. As a practical matter, this means that the ability of citizens to employ various judgmental criteria depends on the larger political debate, and accessibility provides the connecting link between environmentally supplied political messages and the terms and criteria of individual political choice. PMID- 15795131 TI - Emotions and cooperation in economic games. AB - In this paper, we examine decisions to cooperate in economic games. We investigate which payoffs give players the greatest pleasure and whether the pleasure they feel about payoffs predicts their decisions to cooperate. To do this, we modify the ultimatum and dictator games by asking players to consider a fixed set of offers and report their preferences over all offers. Players also report the pleasure they imagine feeling from each possible payoff. Results show that players differ in the extent to which they derive pleasure from fairness or greediness. They also differ in the extent to which their choices depend on what we call "strategic" and "non-strategic" pleasure. Strategic pleasure is the expected pleasure of offers, whereas non-strategic pleasure is the pleasure of accepted payoffs. Players whose pleasure primarily depends on larger payoffs tend to make fair offers in the ultimatum game and selfish offers in the dictator game. They maximize strategic pleasure in the ultimatum game and non-strategic pleasure in the dictator game. Players who derive greater pleasure from fairness tend to act fairly in both games. These players maximize non-strategic pleasure. Brain imaging studies should address the question of whether the observed differences in pleasure and preference are systematically linked to differences in neurological activation. PMID- 15795132 TI - Prospect theory on the brain? Toward a cognitive neuroscience of decision under risk. AB - Most decisions must be made without advance knowledge of their consequences. Economists and psychologists have devoted much attention to modeling decisions made under conditions of risk in which options can be characterized by a known probability distribution over possible outcomes. The descriptive shortcomings of classical economic models motivated the development of prospect theory (D. Kahneman, A. Tversky, Prospect theory: An analysis of decision under risk. Econometrica, 4 (1979) 263-291; A. Tversky, D. Kahneman, Advances in prospect theory: Cumulative representation of uncertainty. Journal of Risk and Uncertainty, 5 (4) (1992) 297-323) the most successful behavioral model of decision under risk. In the prospect theory, subjective value is modeled by a value function that is concave for gains, convex for losses, and steeper for losses than for gains; the impact of probabilities are characterized by a weighting function that overweights low probabilities and underweights moderate to high probabilities. We outline the possible neural bases of the components of prospect theory, surveying evidence from human imaging, lesion, and neuropharmacology studies as well as animal neurophysiology studies. These results provide preliminary suggestions concerning the neural bases of prospect theory that include a broad set of brain regions and neuromodulatory systems. These data suggest that focused studies of decision making in the context of quantitative models may provide substantial leverage towards a fuller understanding of the cognitive neuroscience of decision making. PMID- 15795133 TI - Outcome representations, counterfactual comparisons and the human orbitofrontal cortex: implications for neuroimaging studies of decision-making. AB - Recent research suggests that the primate orbitofrontal cortex (OFC) is critical for representations of outcomes of actions and their subsequent impact on the control of behavior. In parallel, a recent theory of decision-making called decision affect theory (Mellers, Schwartz, and Ritov, Psychological Science, 1997) emphasizes the role of anticipated affective impact of outcomes in guiding choices, and the effects of comparisons with alternative outcomes (i.e., counterfactual effects). In the context of decision affect theory, we present results from two event-related functional MRI experiments consistent with two hypotheses regarding the role of the human OFC in guiding behavior through outcome representation: (1) counterfactual effects are manifested in the human OFC during expectation of outcomes, such that the anticipated affective impact of outcomes is modulated by the nature of the various possible alternative outcomes; (2) a regional specialization exists in the human prefrontal cortex, such that affective impact of potential negative outcomes of actions is represented mainly by the lateral areas of the OFC, while areas situated progressively more medial and dorsal on the ventral and medial PFC are specifically involved in representing the impact of positively valenced outcomes. We also discuss some of the implications that these hypotheses have for neuroimaging studies of reward processing and decision-making, and for studies of neuropsychiatric disorders in which these processes are thought to be disturbed. PMID- 15795134 TI - Functional connectivity with anterior cingulate and orbitofrontal cortices during decision-making. AB - Recent neuroscience research is beginning to discover the brain regions involved in decision-making under uncertainty, but little is known about whether or how these regions functionally interact with each other. Here, we used event-related functional magnetic resonance imaging to examine both changes in overall activity and changes in functional connectivity during risk-taking. Results showed that choosing high-risk over low-risk decisions was associated with increased activity in both anterior cingulate and orbitofrontal cortices. Connectivity analyses revealed that largely distinct, but somewhat overlapping, cortical and subcortical regions exhibited significant functional connectivity with anterior cingulate and orbitofrontal cortices. Additionally, connectivity with the anterior cingulate in some regions, including the orbitofrontal cortex and nucleus accumbens, was modulated by the decision participants chose. These findings (1) elucidate large networks of brain regions that are functionally connected with both anterior cingulate and orbitofrontal cortices during decision making and (2) demonstrate that the roles of orbitofrontal and anterior cingulate cortices can be functionally differentiated by examining patterns of connectivity. PMID- 15795135 TI - Sustained neural activity associated with cognitive control during temporally extended decision making. AB - Decision making has both cognitive and affective components, but previous neuroimaging studies in this domain predominantly have focused on affect and reward. The current study examined a decision-making paradigm that placed strong demands on cognitive control processes by making reward payoffs contingent upon decision-making history. Payoffs were maximized by choosing the option that, paradoxically, was associated with a lower payoff on the immediate trial. Temporal integration requirements were manipulated by varying, across conditions, the window of previous trials over which the reward function was calculated. The cognitive demands of the task were hypothesized to engage neural systems responsible for integrating and actively maintaining actions and outcomes over time and the top-down biasing of response selection. Brain activation was monitored with functional magnetic resonance imaging (fMRI) using a mixed-blocked and event-related design to extract both transient and sustained neural responses. A network of brain regions commonly associated with cognitive control functions, including bilateral prefrontal cortex (PFC), bilateral parietal cortex, and medial frontal cortex, showed selectively sustained activation during the task. Increasing temporal integration demands led to a shift from transient to sustained activity in additional regions, including right hemisphere dorsolateral and frontopolar PFC. These results demonstrate the contribution of cognitive control mechanisms to temporally extended decision-making paradigms and highlight the benefits of decomposing activation responses into sustained and transient components. PMID- 15795136 TI - The dark side of emotion in decision-making: when individuals with decreased emotional reactions make more advantageous decisions. AB - Can dysfunction in neural systems subserving emotion lead, under certain circumstances, to more advantageous decisions? To answer this question, we investigated how individuals with substance dependence (ISD), patients with stable focal lesions in brain regions related to emotion (lesion patients), and normal participants (normal controls) made 20 rounds of investment decisions. Like lesion patients, ISD made more advantageous decisions and ultimately earned more money from their investments than the normal controls. When normal controls either won or lost money on an investment round, they adopted a conservative strategy and became more reluctant to invest on the subsequent round, suggesting that they were more affected than lesion patients and ISD by the outcomes of decisions made in the previous rounds. PMID- 15795137 TI - Cardiac concomitants of performance monitoring: context dependence and individual differences. AB - Feedback processing is an important aspect of cognitive control and decision making. Several studies have shown that heart rate slows following feedback that indicates incorrect performance or loss of money. The current study was the first to investigate (1) whether this slowing reflects an evaluation of the valence of the outcome or a system that indicates that the feedback contains informative value, (2) whether the slowing is determined by the value of the outcome relative to the range of possible outcomes, and (3) whether highly anxious individuals have a hypersensitive feedback monitoring system. The results showed that heart rate only slows when the feedback is performance based. The information provided by negative feedback is processed in a context-sensitive manner, suggesting that heart rate slowing following feedback reflects a signal associated with informative value for subsequent performance adjustment. Highly anxious individuals showed larger heart rate slowing in response to feedback indicating high stakes, but they failed to respond to feedback in a context-sensitive manner. These results were interpreted to suggest that anxious individuals are generally more sensitive to performance outcomes. Heart rate changes following informative feedback proved to be a sensitive index of component processes associated with performance monitoring. PMID- 15795138 TI - Neuroimaging of marijuana smokers during inhibitory processing: a pilot investigation. AB - Neuropsychological investigations of substance abusers have reported impairments on tasks mediated by the frontal executive system, including functions associated with behavioral inhibition and decision making. The higher order or executive components which are involved in decision making include selective attention and short term storage of information, inhibition of response to irrelevant information, initiation of response to relevant information, self-monitoring of performance, and changing internal and external contingencies in order to "stay the course" towards the ultimate goal. Given the hypothesized role of frontal systems in decision making and the previous evidence that executive dysfunctions and structural brain changes exist in subjects who use illicit drugs, we applied fMRI and diffusion tensor imaging (DTI) techniques in a pilot investigation of heavy cannabis smokers and matched control subjects while performing a modification of the classic Stroop task. Marijuana smokers demonstrated significantly lower anterior cingulate activity in focal areas of the anterior cingulate cortex and higher midcingulate activity relative to controls, although both groups were able to perform the task within normal limits. Normal controls also demonstrated increased activity within the right dorsolateral prefrontal cortex (DLPFC) during the interference condition, while marijuana smokers demonstrated a more diffuse, bilateral pattern of DLPFC activation. Similarly, although both groups performed the task well, marijuana smokers made more errors of commission than controls during the interference condition, which were associated with different brain regions than control subjects. These findings suggest that marijuana smokers exhibit different patterns of BOLD response and error response during the Stroop interference condition compared to normal controls despite similar task performance. Furthermore, DTI measures in frontal regions, which include the genu and splenium of the corpus callosum and bilateral anterior cingulate white matter regions, showed no between group differences in fractional anisotropy (FA), a measure of directional coherence within white matter fiber tracts, but a notable increase in trace, a measure of overall isotropic diffusivity in marijuana smokers compared to controls. Overall, results from the present study indicate significant differences in the magnitude and pattern of signal intensity change within the anterior cingulate and the DLPFC during the Stroop interference subtest in chronic marijuana smokers compared to normal controls. Furthermore, although chronic marijuana smokers were able to perform the task reasonably well, the functional activation findings suggest they utilize different cortical processes from the control subjects in order to do so. Findings from this study are consistent with the notion that substance abusers demonstrate evidence of altered frontal neural function during the performance of tasks that involve inhibition and performance monitoring, which may affect the ability to make decisions. PMID- 15795139 TI - Risky decision making and the anterior cingulate cortex in abstinent drug abusers and nonusers. AB - Risky decision making is a hallmark behavioral phenotype of drug abuse; thus, an understanding of its biological bases may inform efforts to develop therapies for addictive disorders. A neurocognitive task that measures this function (Rogers Decision-Making Task; RDMT) was paired with measures of regional cerebral perfusion to identify brain regions that may underlie deficits in risky decision making in drug abusers. Subjects were abstinent drug abusers (> or =3 months) and healthy controls who underwent positron emission tomography scans with H(2)(15)O. Drug abusers showed greater risk taking and heightened sensitivity to rewards than control subjects. Both drug abusers and controls exhibited significant activations in a widespread network of brain regions, primarily in the frontal cortex, previously implicated in decision-making tasks. The only significant group difference in brain activation, however, was found in the left pregenual anterior cingulate cortex, with drug abusers exhibiting less task-related activation than control subjects. There were no significant correlations between neural activity and task performance within the control group. In the drug abuse group, on the other hand, increased risky choices on the RDMT negatively correlated with activation in the right hippocampus, left anterior cingulate gyrus, left medial orbitofrontal cortex, and left parietal lobule, and positively correlated with activation in the right insula. Drug abuse severity was related positively to right medial orbitofrontal activity. Attenuated activation of the pregenual ACC in the drug abusers relative to the controls during performance on the RDMT may underlie the abusers' tendency to choose risky outcomes. PMID- 15795140 TI - Decision making in pathological gambling: a comparison between pathological gamblers, alcohol dependents, persons with Tourette syndrome, and normal controls. AB - Decision making deficits play an important role in the definition of pathological gambling (PG). However, only few empirical studies are available regarding decision making processes in PG. This study therefore compares decision making processes in PG and normal controls in detail using three decision making tasks examining general performance levels on these tasks as well as feedback processing using reaction time analyses. To investigate the specificity of decision making deficits in PG, a substance dependence group (alcohol dependence; AD) and an impulse control disordered group (Tourette syndrome; TS) were included. The PG group (n = 48), AD group (n = 46), TS group (n = 47), and a normal control (NC) group (n = 49) were administered (1) the Iowa Gambling Task (IGT), an ecologically valid gambling task; (2) the Card Playing Task, a task measuring perseveration for reward; and (3) a Go/No-Go discrimination task, a task measuring reward and response cost sensitivity. The PG group showed a diminished performance on all tasks and deficient feedback processing as compared to the NC group on the IGT and the Card Playing Task. In general, performance measures were not associated with levels of comorbidity or with self-reported motivational measures. For the larger part, deficiencies in decision making processes in the PG group were also present in the AD group, but not in the TS group. Subgroup analyses revealed larger decision making deficits in pathological slot machine gamblers than in pathological casino gamblers. Deficits in decision making and feedback processing in PG should be addressed in treatment and incorporated more explicitly in theoretical models of PG. PMID- 15795141 TI - Fifty years of confidential enquiries into maternal deaths in the United Kingdom: should anaesthesia celebrate or not? PMID- 15795142 TI - A prospective observational study of the use of the Proseal laryngeal mask airway for postpartum tubal ligation. AB - BACKGROUND: Though controversial, the risk of pulmonary aspiration during general anaesthesia in the immediate postpartum period appears low. The efficacy of the Proseal laryngeal mask airway was studied prospectively in a group of patients undergoing postpartum tubal ligation. METHODS: The Proseal laryngeal mask airway was employed for airway management in 90 fasted patients undergoing tubal ligation via minilaparotomy at least 8 h after normal vaginal delivery (mean 36.5, range 8-96 h). Gastric volume and pH were measured, using aspiration through a gastric tube. RESULT: Proseal laryngeal mask airway insertion was successful in all patients, requiring one attempt in 75 patients (83%). The median (range) leak pressure was 35 (23-40) cmH2O. Twenty-two patients (25%) had a leak pressure of 40 cmH2O or greater. Gastric tube placement was successful in all patients, described as easy in 79 (87%), and difficult in 11 (13%). The mean initial volume of gastric aspirate was 10.7 (0-64) mL and the final volume 15.6 (0-71) mL. The mean pH of the gastric aspirate was 2.6 (1.2-6.6). There were no incidents of suspected fluid regurgitation or aspiration, but two patients required intubation during surgery. Ten patients (11.1%) complained of sore throat in the recovery room, nine of which were described as mild. All patients reported being satisfied with their anaesthesia. CONCLUSION: The Proseal laryngeal mask airway provides an effective airway for general anaesthesia in fasted patients undergoing tubal ligation from 8 h after normal vaginal delivery. While the safety of an unprotected airway in this population remains uncertain, this study suggested a low risk of regurgitation, especially in the first 24 h post partum. PMID- 15795143 TI - Obstetric epidural test doses: a survey of UK practice. AB - BACKGROUND: With the advent of low-dose epidural analgesia in labour, the content of the test dose has once again become the subject of debate. METHOD: A postal survey of 500 members of the Obstetric Anaesthetists' Association was conducted in 1999-2000, assessing the use of test doses during epidurals in labour and for caesarean section. RESULTS: There was a 67% response rate. Test doses are used in labour, at elective caesarean section and before epidural top-up for emergency caesarean section, by 90%, 93% and 37%, respectively. There was large variation in both drugs and doses. During labour, doses of bupivacaine range from 3 to 20 mg and of lidocaine 15 to 90 mg. There has been a three-fold increase in the use of low-dose local anaesthetic test doses since a previous national survey in 1997. The size of local anaesthetic test doses used at caesarean section is also variable. Epinephrine is used in 5% of labour, 14% of elective and 34% of emergency caesarean sections. Signs and symptoms that are commonly sought after test doses include somatic motor block, blood pressure change, sensory effect and symptoms from systemic local anaesthetic. The effect of the test dose is usually assessed after 5 min. CONCLUSION: There is no consensus about the nature of the ideal test dose in obstetric anaesthesia. There is a trend to use less concentrated test doses during labour. Doses that risk a high block if given spinally are still used. Epinephrine, aspiration testing and cardiovascular monitoring are uncommon. PMID- 15795144 TI - The in vitro effects of clonidine and dexmedetomidine on human myometrium. AB - BACKGROUND: alpha(2)-adrenergic agonists have been used extensively in the field of anaesthesia. Their direct effect on the human myometrium was investigated in this in vitro study, as this may have clinical repercussions in obstetric anaesthesia. METHOD: Strips of pregnant human myometrium obtained from six individuals at elective caesarean section were mounted on the Mulvany myograph in Krebs solution to which increasing concentrations of clonidine and dexmedetomidine (1x10(-11) to 1x10(-6) g/mL) were added. RESULTS: Dexmedetomidine increased uterine contractility at simulated clinical plasma concentrations (1x10(-9) g/mL). These effects were seen with clonidine only at much higher tissue bath concentrations (1x10(-7) g/mL). CONCLUSION: The effect of dexmedetomidine on human myometrium has profound implications in obstetric anaesthesia and needs further clinical investigation. PMID- 15795145 TI - Maternal and fetal outcome of anaesthesia for caesarean delivery in preeclampsia/eclampsia in Enugu, Nigeria: a retrospective observational study. AB - BACKGROUND: Maternal mortality, for which preeclampsia is a major cause, is a problem in Nigeria. Accurate data are available for caesarean sections in the University of Nigeria Teaching Hospital, Enugu. We therefore studied the outcome of caesarean section among these high-risk patients. METHOD: We conducted a retrospective survey of hospital records of patients with preeclampsia/eclampsia who had caesarean delivery in this unit over a four-year span from July 1998 to June 2002. RESULTS: There were 3926 deliveries and 4036 births (3611 live births), with 898 women (23%) delivered by caesarean section. Of these, 125 (14%) had preeclampsia/eclampsia, 103 (82.4%) presenting for emergency caesarean delivery and 22 (17.6%) elective. General anaesthesia was used in 116 patients (92.8%) and spinal in nine. The major indications for surgery were severe preeclampsia/eclampsia in patients with unfavourable cervix (68%), fetal distress/intrauterine growth restriction (7.2%) and previous caesarean section (6.4%). There were six maternal deaths, all with general anaesthesia, giving a case fatality rate of 5.2% of general anaesthetics or 4.8% of caesarean deliveries. The cause of death was anaesthetic in three patients, cerebrovascular accident and pulmonary oedema in two and intraoperative haemorrhage in one. There were 13 stillbirths and 10 neonatal deaths. CONCLUSION: Maternal and fetal mortality were high. Poverty, late presentation, lack of equipment and inexperienced management were major contributory factors. Use of spinal anaesthesia should be encouraged in view of recent favourable reviews and cheaper cost. PMID- 15795147 TI - Intrathecal epinephrine in combined spinal-epidural analgesia for labor: dose response relationship for epinephrine added to a local anesthetic-opioid combination. AB - BACKGROUND: The purpose was to investigate the dose-response relationship for intrathecally administered epinephrine added to a local anesthetic-opioid combination in combined spinal-epidural analgesia for labor, in order to evaluate analgesia and side-effects. PATIENTS AND METHODS: The subjects were 100 consecutive ASA I or II parturients at 37 weeks' gestation, who received combined spinal-epidural analgesia during labor. Each woman was randomly assigned to one of five groups that received 2-mL volumes of different spinal solutions. The control group received an intrathecal injection of bupivacaine 2.5 mg and fentanyl 25 microg only. The others received epinephrine 12.5, 25, 50 or 100 microg added to this intrathecal regimen. Maternal arterial pressure, heart rate and pain scores were recorded before and 5, 10, 15 and 30 min after intrathecal injection. Level of sensory blockade, motor blockade score, duration of intrathecal analgesia, side effects, fetal heart rate, and 1- and 5-min Apgar scores were also assessed. RESULTS: Compared to the control group, all four epinephrine groups had significantly longer duration of intrathecal analgesia, but the durations were similar. The frequencies of side effects were similar in all five groups. CONCLUSION: The results suggest that adding epinephrine to a combination of standard intrathecal doses of bupivacaine and fentanyl in combined spinal-epidural analgesia for labor significantly prolongs spinal analgesia. Of the four epinephrine doses tested, the lowest one (12.5 microg) was optimal for this clinical setting. PMID- 15795146 TI - Comparison of continuous background infusion plus demand dose and demand-only parturient-controlled epidural analgesia (PCEA) using ropivacaine combined with sufentanil for labor and delivery. AB - BACKGROUND: Using ropivacaine combined with sufentanil, we determined the analgesic efficacy of parturient-controlled epidural analgesia (PCEA) with or without (demand-only PCEA) continuous background infusion in reducing labor pain in 66 parturients. METHODS: After placement of the epidural catheter and administration of an initial bolus containing ropivacaine 16 mg and sufentanil 10 microg, parturients were prospectively randomized into two groups. The PCEA solution consisted of ropivacaine 0.16% plus sufentanil 0.5 microg/mL. Parturients with PCEA plus continuous background infusion received 4 mL/h plus an hourly maximum of three 4-mL boluses on demand (lock-out time 20 min); parturients with demand-only PCEA received an hourly maximum of four 4-mL boluses (lock-out time 15 min) of anesthetic solution. Pain scores (VAS 0-100 mm), drug doses administered, duration of labor, sensory and motor epidural block characteristics, maternal satisfaction, neonatal outcome and adverse events were determined. RESULTS: Both regimens provided excellent parturients' satisfaction and pain relief. However, periods of VAS scores>40 mm during all stages of labor were significantly more frequent in parturients receiving demand-only PCEA (22.4%) compared to parturients receiving PCEA plus continuous background infusion (7.5%, P=0.0011). Drug doses administered, duration of PCEA, labor and delivery, epidural block characteristics, neonatal outcome and adverse events did not differ between groups. CONCLUSION: Under the conditions of the study, PCEA plus continuous background infusion was more effective than demand-only PCEA in treating labor pain without increasing consumption of anesthetic solution. PMID- 15795148 TI - What's new and novel in obstetric anesthesia? Contributions from the 2003 scientific literature. AB - THE PREGNANT PATIENT: Age; maternal disease; prophylactic antibiotics; gastroesophageal reflux; obesity; starvation; genotyping; coagulopathy; infection; substance abuse; altered drug responses in pregnancy; physiological changes of pregnancy. THE FETUS: Fetal monitoring; intrauterine surgery. THE NEWBORN: Breastfeeding; maternal infection, fever, and neonatal sepsis evaluation. OBSTETRIC COMPLICATIONS: Embolic phenomena; hemorrhage; preeclampsia; preterm delivery. OBSTETRIC MANAGEMENT: External cephalic version and cervical cerclage; elective cesarean delivery; fetal malpresentation; vaginal birth after cesarean delivery; termination of pregnancy. OBSTETRIC ANESTHESIA: Analgesia for labor and delivery; anesthesia for cesarean delivery; anesthesia for short obstetric operations; complications of anesthesia. MISCELLANEOUS: Consent; ethics; history; labor support; websites/books/leaflets/journal announcements. PMID- 15795149 TI - The National Institute of Clinical Excellence (NICE) guidelines for caesarean sections: implications for the anaesthetist. PMID- 15795150 TI - Intracranial subdural hematoma after spinal anesthesia. AB - Intracranial subdural hematoma is an exceptionally rare but life-threatening complication of spinal anesthesia. We report a case of intracranial subdural hematoma following spinal anesthesia for cesarean section in a 27-year-old woman. She developed a diffuse headache after surgery with a blood pressure of 220/140 mm Hg which was followed by generalized seizure activity. Her blood pressure remained high after medication with diazepam, nifedipine and magnesium sulfate. She remained unconscious with a Glasgow coma scale of 5. The cranial tomography revealed a subdural hematoma with diffuse cerebral edema and cerebral tentorial herniation. When a patient complains of postdural puncture headache and then has seizure activity, one should consider alternative diagnoses, including that of a subdural hematoma, and carry out a careful examination, including magnetic resonance imaging or computerized tomography scan. PMID- 15795151 TI - A case of late postpartum seizures after epidural analgesia. AB - We present the case of a 29-year-old female who in her third pregnancy developed seizures seven days postpartum. The only symptoms preceding the seizures were headache and visual disturbance. There was no evidence of preeclampsia antepartum or postpartum. All clinical investigations were normal including computerised tomography, magnetic resonance imaging and lumbar puncture. All symptoms resolved within 24 h of the first seizure. In this case report we highlight the difficulties in establishing the cause of postpartum seizures. PMID- 15795152 TI - Anaesthetic considerations in a parturient with critical coronary artery disease and a drug-eluting stent presenting for caesarean section. AB - A parturient presented with her first symptoms of coronary artery disease at 18 weeks' gestation. Following an angiogram, a drug-eluting stent was inserted, resulting in resolution of her symptoms. The patient was prescribed anti-platelet medication including clopidogrel. She was delivered by elective caesarean section at 35 weeks under general anaesthesia. The anaesthetic management is discussed and a review of the literature presented. PMID- 15795153 TI - Spontaneous common bile duct rupture in pregnancy. AB - This report describes how a rare condition in pregnancy can present with features resembling more common diagnoses in the unwell obstetric patient. Our patient presented in late pregnancy initially with clinical features similar to mild preeclampsia, with proteinuric hypertension, epigastric pain and mildly deranged liver function tests. She went on to develop signs of acute abdomen associated with a persistent tachycardia and hypotension; there was evidence of fetal compromise. Following resuscitation, emergency caesarean section was performed and evolved into a laparotomy. The diagnosis was not clear clinically at this stage, but a retrospective amylase sample suggested acute pancreatitis. There was no clinical evidence of biliary perforation, but ongoing clinical suspicion prompted a CT of the abdomen. This proved unhelpful and our diagnostic dilemma was only resolved by endoscopic retrograde cholangiopancreatography, which demonstrated a biliary leak. A stent was placed with subsequent improvement in the patient's condition. With anaesthetists increasingly involved in the multi disciplinary management of acutely sick obstetric patients, our case highlights the need for every practitioner involved in such cases to assess each individual fully, and be constantly aware that the diagnosis might not be obvious. PMID- 15795154 TI - Failed regional anesthesia with reduced spinal bupivacaine dosage in a parturient with achondroplasia presenting for urgent cesarean section. AB - A 36-year-old patient with a history of previous back surgery, asthma, latex allergy and achondroplasia presented for urgent cesarean delivery at 31 weeks' gestation for worsening nonimmune fetal hydrops. The fetus was diagnosed with trisomy 21 and achondroplasia. Because of the urgent clinical situation, the patient was given a spinal anesthetic, which required supplemental intravenous sedation after delivery of the fetus. This case report discusses the controversies in anesthetic management of this complicated patient and compromised fetus regarding general anesthesia, epidural, spinal and combined spinal-epidural anesthesia. PMID- 15795155 TI - Three spinal anaesthetics for caesarean sections in one patient, two requiring blood patch. PMID- 15795156 TI - Intravenous prostaglandin F2alpha? PMID- 15795157 TI - Role of progenitor cells in transplant arteriosclerosis. AB - To date, chronic transplant dysfunction (CTD) is recognized as the major cause of transplant loss long term after transplantation. CTD has the remarkable histologic feature that the luminal areas of the intragraft arteries become obliterated as a result of occlusive neointima formation. Neointimal lesions contain predominantly vascular smooth muscle cells (VSMCs) and extracellular matrix admixed with inflammatory cells. At the luminal side, neointimal lesions are covered with a monolayer of endothelial cells (ECs). The etiology of transplant arteriosclerosis (TA) is largely unknown, and adequate prevention and treatment protocols are not available. In contrast to the largely accepted "response-to-injury" hypothesis for the development of TA that attributes an important role to graft-derived ECs and VSMCs, recent data indicate that host derived vascular progenitor cells play a major role in the development of TA. The process leading to TA appears to be heterogeneous, and neointimal ECs and VSMCs can be recruited from different sources, possibly depending on the severity and duration of vascular damage. These data suggest a significant role of host derived circulating EC/VSMC progenitor cells, which may be partly bone marrow derived. Circulating vascular progenitor cells are potential targets for therapeutic intervention to ameliorate TA development. Therefore, identification of mediators and cellular mechanisms that promote recruitment of vascular progenitors to sites of injury is warranted to dissect their detrimental and possible beneficial effects in the development of TA. PMID- 15795159 TI - Imaging of atherosclerosis -- can we predict plaque rupture? AB - Rupture of so-called vulnerable or unstable atherosclerotic lesions is responsible for a significant proportion of myocardial infarcts and strokes. However, timely identification of such plaques, in order to allow for aggressive local and systemic therapy, remains problematic. In order to address this problem, there is a need to develop techniques that can image the cellular, biochemical, and molecular components that typify the vulnerable plaque. In this article, both techniques that are in current clinical use and those being evaluated in clinical trials are reviewed with regard to their ability to identify unstable lesions at risk of rupture. PMID- 15795158 TI - Signaling to translational control pathways: diversity in gene regulation in inflammatory and vascular cells. AB - The expression of a subset of genes is strongly controlled at translational checkpoints, a major mechanism of posttranscriptional regulation. Inflammatory and vascular cells receive outside-in signals to specialized pathways that regulate translation of specific messenger RNAs in a transcript-specific fashion and thereby influence key features of cellular phenotype. These pathways and the expression of proteins that they control may be dysregulated in cardiovascular diseases and are therapeutic targets. PMID- 15795160 TI - Signaling mechanisms in cerebral vasospasm. AB - The elusive nature of events that sustain cerebral vasospasm after subarachnoid hemorrhage resulting from a ruptured aneurysm presents major challenges in designing effective therapies for this frequently devastating condition. Protracted cerebral artery constriction entails several dynamic components in intracellular signaling events initiated by endothelial factors, products of hemolysate, and numerous kinases, as well as increased intracellular Ca(2+). The rationale for potential treatment modalities and their efficacy are discussed in this brief review. PMID- 15795161 TI - Regulation of the voltage-gated cardiac sodium channel Nav1.5 by interacting proteins. AB - Na(v)1.5, the major cardiac voltage-gated Na(+) channel, plays a central role in the generation of the cardiac action potential and in the propagation of electrical impulses in the heart. Its importance for normal heart function has been recently exemplified by reports of numerous mutations found in the gene SCN5A--which encodes Na(v)1.5--in patients with various pathologic cardiac phenotypes, indicating that even subtle alterations of Na(v)1.5 cell biology and function may underlie human diseases. Similar to other ion channels, Na(v)1.5 is most likely part of dynamic multiprotein complexes located in the different cellular compartments. This review focuses on five intracellular proteins that have been recently reported to directly bind to and contribute to the regulation of Na(v)1.5: ankyrin proteins, fibroblast growth factor homologous factor 1B, calmodulin, Nedd4-like ubiquitin-protein ligases, and syntrophin proteins. PMID- 15795162 TI - The potential of aptamers as anticoagulants. AB - Useful additional options for anticoagulant therapy have been introduced over the last 15 years, including low-molecular-weight heparins and direct thrombin inhibitors. Despite these impressive advances, a need for safer effective anticoagulants remains. Aptamers represent a therapeutic modality that has the potential to address this unmet need. Aptamers are small nucleic acid molecules that function as direct protein inhibitors, much like monoclonal antibodies. Aptamers are delivered by parenteral administration, can be formulated to possess a very short or sustained half-life, and are purported to be nonimmunogenic. Perhaps most relevant to the development of safer anticoagulant therapies, recent studies have shown that antidotes can be rationally designed to control the pharmacologic effects of aptamers in vivo, paving the way for a new class of antidote-controlled therapeutics. This review discusses the limitations of current anticoagulant therapies, the properties of aptamers and how these properties can be exploited to address the unmet needs within this therapeutic class, and the progress to date in developing new aptamer-based anticoagulant therapies. PMID- 15795163 TI - Induction of apoptosis in tumor cells by naturally occurring sulfur-containing compounds. AB - Chemoprevention is regarded as one of the most promising and realistic approaches in the prevention of human cancer. Among naturally occurring products, sulfur containing compounds (OSCs), especially garlic compounds (GCs) and isothiocyanates (ITCs), represent two important and promising chemopreventive families because of their potent chemopreventive effects in various in vivo and in vitro models. In recent years, numerous investigations have shown that sulfur containing compounds induce apoptosis in multiple cell lines and experimental animals. In the course of apoptosis induction by GCs and ITCs, multiple signal transduction pathways and apoptosis intermediates are modulated. In particular, modulation of MAPKs and production of reactive oxygen species (ROS) seem to play pivotal roles in apoptosis induction by most GCs and ITCs. However, the role of P53 is still controversial. Based on present knowledge, GCs and ITCs may target not only the metabolism of carcinogens but also apoptosis signaling molecules. The effects of ITCs and GCs at multiple points of cancer development make these compounds highly promising candidates in cancer chemoprevention. However, the mechanisms of their anticancer effects are not fully understood, and further studies are required, especially to elucidate the role of cell-death receptors (the extrinsic pathway) and whether these agents induce apoptotic effects in non tumor cells. PMID- 15795164 TI - Lead toxicity through the leadzyme. AB - Lead is one of the most dangerous toxic agents for all living organisms. In humans, elevated levels of lead have been linked to a number of disorders for which various molecular mechanisms have been proposed. However, none of them has been fully understood. It has also been known for several years that at micromolar concentrations lead can bind a unique RNA motif and catalyze a site specific hydrolysis of the polyribonucleotide chain. This motif, called leadzyme, may be one of the major targets for lead within the cell, and it can cleave various cellular RNAs. A search of GenBank revealed the sequences that can potentially fold into the structure containing the leadzyme motif and that they are rather common in eukaryotic genomes. We found that the domain occurs with a high frequency in human mRNA sequences. Thus, the leadzyme nucleolytic properties should be considered as a possible mechanism for destruction of RNA within a cell. In particular, targeting of the RNA scaffold of ribosomes or spliceosomes may explain lead-mediated toxicity leading to cell death. PMID- 15795165 TI - DNA-protein crosslinks: their induction, repair, and biological consequences. AB - The covalent crosslinking of proteins to DNA presents a major physical challenge to the DNA metabolic machinery. DNA-protein crosslinks (DPCs) are induced by a variety of endogenous and exogenous agents (including, paradoxically, agents that are known to cause cancer as well as agents that are used to treat cancer), and yet they have not received as much attention as other types of DNA damage. This review summarizes the current state of knowledge of DPCs in terms of their induction, structures, biological consequences and possible mechanisms of repair. DPCs can be formed through several different chemistries, which is likely to affect the stability and repair of these lesions, as well as their biological consequences. The considerable discrepancy in the DPC literature reflects both the varying chemistries of this heterogeneous group of lesions and the fact that a number of different methods have been used for their analysis. In particular, research in this area has long been hampered by the inability to chemically define these lesions in intact cells and tissues. However, the emergence of proteomics as a tool for identifying specific proteins that become crosslinked to DNA has heralded a new era in our ability to study these lesions. Although there are still many unanswered questions, the identification of specific proteins crosslinked to DNA should facilitate our understanding of the down-stream effects of these lesions. PMID- 15795166 TI - Genetic toxicity and carcinogenicity studies of glutaraldehyde--a review. AB - Glutaraldehyde is a high production volume chemical that is highly reactive and has many medical and industrial uses. The majority of human exposures are via inhalation, but the exposure is not widespread. It has been extensively tested for genetic activity in vitro and in vivo, and there is disagreement in the literature with regard to glutaraldehyde's genetic activity. Glutaraldehyde produced DNA damage in bacteria and some cultured mammalian cell systems. In vitro, it was mutagenic in Salmonella and E. coli, produced inconsistent positive responses in mammalian cells, weak and inconsistent responses in chromosome aberration and SCE studies, and did not induce transformation in cultured SHE cells. In vivo, inhalation of glutaraldehyde induced cell proliferation in nasal tissue in rats and mice, but DNA damage and UDS were not induced at these sites in rats. Chromosome aberrations in bone marrow cells were reported in only one of eight studies using rats and mice, micronuclei were not induced in bone marrow cells of mice, and dominant lethal mutations were not induced in mice. Glutaraldehyde did not induce cell transformation in SHE cells in vitro. Bone marrow hyperplasia and low, but statistically significant, levels of leukemia were seen in one chronic drinking water study in rats, but not in a chronic inhalation study in rats or two chronic inhalation studies in mice. PMID- 15795167 TI - Reliable manual segmentation of the frontal, parietal, temporal, and occipital lobes on magnetic resonance images of healthy subjects. AB - BACKGROUND: It is a challenge to reliably measure the lobar volumes from magnetic resonance imaging (MRI) data. OBJECTIVE: Description of a landmark-based method for volumetric segmentation of the brain into the four cerebral lobes from MR images. METHOD: The segmentation method relies on a combination of anatomical landmarks and geometrical definitions. The first step, described previously, is a segmentation of the four lobes on the surface of the brain. The internal borders between the lobes are defined on the axial slices of the brain. The intra- and inter- rater reliability was determined from the MRI scans of a group of 10 healthy control subjects measured by 2 independent raters. RESULTS: The intra rater relative error (and intra-class correlation coefficient) of the lobar volume measures ranged from 0.81% to 3.85% (from 0.97 to 0.99). The inter-rater relative error (and intra-class correlation coefficient) ranged from 0.55% to 3.09% (from 0.94 to 0.99). CONCLUSION: This technique has been shown to have high intra- and inter-rater reliability. The current method provides a method to obtain volumetric estimates of the 4 cerebral lobes. PMID- 15795168 TI - Adult CNS explants as a source of neural progenitors. AB - Adult neural progenitors have been isolated from diverse regions of the CNS using methods which primarily involve the enzymatic digestion of tissue pieces; however, interpretation of these experiments can be complicated by the loss of anatomical resolution during the isolation procedures. We have developed a novel, explant-based technique for the isolation of neural progenitors. Living CNS regions were sectioned using a vibratome and small, well-defined discs of tissue punched out. When cultured, explants from the cortex, hippocampus, cerebellum, spinal cord, hypothalamus, and caudate nucleus all robustly gave rise to proliferating progenitors. These progenitors were similar in behaviour and morphology to previously characterised multipotent hippocampal progenitor lines. Clones from all regions examined could proliferate from single cells and give rise to secondary neurospheres at a low but consistent frequency. Immunostaining demonstrated that clonal cortical progenitors were able to differentiate into both neurons and glial cells, indicating their multipotent characteristics. These results demonstrate it is possible to isolate anatomically resolved adult neural progenitors from small amounts of tissue throughout the CNS, thus, providing a tool for investigating the frequency and characteristics of progenitor cells from different regions. PMID- 15795170 TI - A novel method of detecting rCBF with laser-Doppler flowmetry without cranial window through the skull for a MCAO rat model. AB - BACKGROUND AND PURPOSE: In a rat model of middle cerebral artery occlusion (MCAO) with intraluminal technique, lesion volume and its reproducibility vary among laboratories. Although laser-Doppler flowmetry (LDF) is useful to optimize the reliability, conventional methods require a craniotomy and special apparatus. The purpose of this study was to evaluate a novel approach for LDF monitoring of rCBF through lateral aspect of the skull without a craniotomy. METHODS: SD rats were subjected to 45 min of MCAO using an intraluminal thread. MCAO was achieved by an examiner who had been trained 4 weeks for making the model with no LDF monitoring (Group-1, n = 12), while in the other group, the same examiner induced MCAO using a novel approach of LDF monitoring (Group-2, n = 12). rCBF was detected through an LDF probe attached to the lateral aspect of the skull. The survival rate and the infarct volume were estimated for comparison between the two groups 2 days after MCAO. RESULTS: The mortality rate was 25% in Group-1 and 0% in Group-2. The lesion volume of the cortex in Group-2 was 167.21 +/- 48.54 mm(3) (mean +/- SD), which was larger than that in Group-1 (112.77 +/- 36.03 mm(3), P = 0.026). The coefficient variation of the lesion volume was smaller in Group-2 (29%) than in Group-1 (35%), indicating better reproducibility of the lesion volume in Group-2 than in Group-1. CONCLUSIONS: The approach of LDF monitoring through the lateral aspect of skull was useful for making large consistent infarct with reducing intraanimal variability and unexpected animal death for rat MCAO model. PMID- 15795169 TI - Electrocorticographic and deep intracerebral EEG recording in mice using a telemetry system. AB - Telemetric EEG recording plays a crucial role in the neurological characterization of various transgenic mouse models giving valuable information about epilepsies and sleep disorders in humans. In the past different experimental approaches have been described using tethered systems and jacket systems containing recorders. A main disadvantage of these is their sometimes unphysiological, restraining character. Telemetric EEG recording overcomes most of these disadvantages and allows precise and highly sensitive measurement under various physiological and pathophysiological conditions and different stages of consciousness, as during seizure activity and different sleep stages. Here we present the first contiguous, detailed description of a successful and quick technique for intraperitoneal implantation or subcutaneous pouch implantation of a radiofrequency transmitter in mice and subsequent lead placement in both epidural and deep intracerebral position. Preoperative preparation of the mice, suitable anesthesia, as well as postoperative treatment including pain management are described in detail to provide optimal postoperative recovery. Finally, we display examples of electrocorticograms and deep intracerebral recordings, present strategies to maximize signal-to-noise ratio, paying special attention to major pitfalls and possible artefacts occurring in telemetric EEG recording in mice. PMID- 15795171 TI - New canine spinal cord injury model free from laminectomy. AB - The present report details the successful development of a model for spinal cord injury (SCI). This model is simple, reproducible, and requires no laminectomy. Development of the model was carried out using fourteen dogs. A balloon catheter was inserted into the extradural space via the intervertebral foramen of each dog, then the balloon was inflated at the L1 level by injection of saline. Six dogs underwent compression with a balloon volume of 1.5 ml, three dogs with a volume of 1.0 ml, and the remaining five dogs were used as uninjured controls. We applied the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale to the dogs. Compression of the spinal cord for 10 min at 1.5 ml produced severe paraplegia (BBB remained zero or one for 6 months following surgery), while compression for the same time interval at 1.0 ml produced moderate paraplegia. Electrophysiological tests showed no hindlimb movement upon stimulation cranial to the site of injury in the 1.5-ml group. The volume of abnormal-intensity lesions in the 1.0-ml group calculated using MR imaging showed no marked changes in either high- or low-intensity lesions after 3 months, whereas in the 1.5-ml group, the low-intensity lesions alone showed a marked increase. Pathological examination of the damaged spinal cord showed the formation of cavities surrounded by scar tissue containing high levels of collagen. These findings closely resembled those of clinical cases. It was concluded that 10 min of balloon compression with a volume of 1.5 ml caused irreversible paraplegia in dogs. PMID- 15795172 TI - Modern anesthesia and peroperative monitoring methods reduce per- and postoperative mortality during transient occlusion of the middle cerebral artery in rats. AB - Mortality and morbidity during and after occlusion of the middle cerebral artery in rats are important confounding factors which may be minimized by improved anesthesia and peroperative monitoring techniques. We describe state of the art techniques for inducing anesthesia, endotracheal intubation, ventilation and monitoring peroperatively in this context. Introducing the subtemporal approach of Tamura et al. in our laboratory 5 years ago, we experienced 25% peroperative and 24 h postoperative rat mortality when performing temporary clipping of the middle cerebral artery. This prompted us to abandon intraperitoneal anesthesia by chloral hydrate and ventilation by tracheotomy in favor of endotracheal intubation and isoflurane anesthesia (1% isoflurane in 30%:70% O(2)/N(2)O). These anesthetic techniques in combination with improved surgical skills have reduced our initial 25% peroperative- and 24 h postoperative mortality to 2.7% (1.8% peroperatively and 0.9% 24 h postoperatively). Furthermore, the following 14 days postoperative mortality rate was 1.8%. A total number of 203 rats have been operated with this method in different studies where a focal reperfusion stroke model combined with extended periods of observations were the cornerstone. PMID- 15795173 TI - Brief stimulation of the peroneal nerve attenuates the exercise pressor reflex in anaesthetised cats. AB - We recently demonstrated that applying capsaicin to the common peroneal nerve, thereby activating small diameter afferent neurons, caused a substantial rise in mean arterial pressure (MAP) and heart rate (HR) that lasted approximately 20 min. In addition, this application of capsaicin transiently attenuated the exercise pressor reflex (EPR). The purpose of the current study was to test the hypothesis that stimulating the peroneal nerve at an intensity that activated both myelinated and unmyelinated axons for a short duration (1 min) causes a similar attenuation of the EPR. Cats were anaesthetised with alpha-chloralose and urethane, the popliteal fossa was exposed, and static contraction was induced by stimulating the tibial nerve. The ipsilateral peroneal nerve was cut and placed on a stimulating electrode. Prior to peroneal nerve stimulation, static contraction of the triceps surae muscle for 1 min increased MAP 48+/-8 mmHg and HR 16+/-3 bpm. Electrical stimulation of the central end of the cut peroneal nerve for 1 min (100 x motor threshold; 40 Hz; 0.1 ms) increased MAP and HR by 62+/-11 mmHg and 28+/-4 bpm, respectively. These increases returned to prestimulation levels within 1 min. Two minutes after the peroneal stimulation was stopped, the EPR was markedly reduced as muscle contraction increased MAP and HR by 20+/-4 mmHg and 7+/-2 bpm, respectively. Repeating the muscle contraction approximately 25 min after peroneal stimulation increased MAP and HR by 38+/-8 mmHg and 12+/-2 bpm, indicating some recovery of the EPR. These results show that brief (1 min) electrical stimulation of afferent neurons in the peroneal nerve attenuates the EPR. This supports the hypothesis that strong activation of small diameter afferent neurons stimulates a nervous system mechanism that diminishes the sensory input from skeletal muscle involved in cardiovascular regulation. PMID- 15795174 TI - Choline acetyltransferase and inducible nitric oxide synthase are increased in myenteric plexus of diabetic guinea pig. AB - Alterations in enzymes in myenteric neurons from ileum were investigated in guinea pigs treated with either the pancreatic beta cell toxin streptozotocin or vehicle. After 5-6 weeks, expressions of choline acetyltransferase, neuronal nitric oxide synthase and inducible nitric oxide synthase were determined in longitudinal and myenteric plexus preparations using indirect immunohistochemistry. In ileum from streptozotocin-treated animals, the density of choline acetyltransferase-immunoreactive nerve fibers within the tertiary plexus and the percent total myenteric neurons expressing inducible nitric oxide synthase were increased, but the percent total myenteric neurons expressing neuronal nitric oxide synthase was not changed. Diabetes resulted in selective alterations in myenteric neurons including an increased density of cholinergic tertiary fibers and percentage of neurons expressing the inducible isoform of nitric oxide synthase. These adaptive changes by myenteric neurons to diabetes may contribute to gastrointestinal dysfunctions associated with diabetes. PMID- 15795175 TI - Paradoxical respiratory sinus arrhythmia in the anesthetized rat. AB - This study examines respiratory sinus arrhythmia (RSA) in the isoflurane anesthetized rat. In fifteen female Sprague-Dawley rats, we recorded continuous ECG and respiratory airflow before and after bilateral vagotomy. RSA was assessed using power spectral analysis and by plotting the normalised changes in heart period as a function of the time during the respiratory cycle. Contrary to descriptions of RSA in conscious rats, we observed in all rats in the current study a 'reversed' pattern of RSA in which heart rate decelerated during inspiration. Elimination of vagal efferent fibres to the heart by vagotomy did not abolish the presence of reversed RSA suggesting that the pattern of heart period variation is not neural, and may be largely mechanical. Vagotomy altered breathing by increasing respiratory period, tidal volume, and the time to peak inspiratory flow. These changes did not alter the magnitude of RSA but reduced the latency period between inspiratory onset and the onset of respiratory related prolongation of heart period. Periods of positive pressure ventilation were associated with reversal of the inspiratory cardiac-deceleration pattern of RSA to resemble the more widely described pattern of inspiratory cardiac acceleration. We conclude that RSA is not a suitable measure of vagal tone during anesthesia in the rat and reiterate the caution that needs to be taken when working with anesthetized experimental models of cardiac control. PMID- 15795176 TI - The influences of p75 neurotrophin receptor and brain-derived neurotrophic factor in the sympathetic innervation of target tissues during murine postnatal development. AB - Post-ganglionic sympathetic neurons express the p75 neurotrophin receptor (p75NTR) and brain-derived neurotrophic factor (BDNF), which together have been implicated in controlling the degree of efferent innervation of peripheral organs [Kohn, J., Aloyz, R.S., Toma, J.G., Haak-Frendscho, M., Miller, F.D. 1999. Functionally Antagonistic Interactions between the TrkA and p75 Neurotrophin Receptors Regulate Sympathetic Neuron Growth and Target Innervation. J. Neurosci. 19, 5393-5408]. To examine this concept further, we developed null mutant mice lacking both p75NTR and BDNF, and assessed whether the loss of this receptor ligand interaction negatively impacts the degree of sympathetic innervation to various target tissues. Between postnatal days 10 and 14, hearts, urinary bladders, kidneys, and submandibular salivary glands were isolated from p75(-/ )/BDNF-/-, p75-/-, BDNF-/-, and wild type siblings. Sympathetic axons were visualized using tyrosine hydroxylase (TH) immunohistochemistry, and TH protein levels were quantified by immunoblotting. Concerning the sympathetic innervation of the heart, urinary bladder and kidneys, no differences were seen in single and double null mutant mice, as compared with their wild type siblings. Sympathetic innervation of the submandibular salivary gland was, however, increased in both p75-/- and p75(-/-)/BDNF-/- mice over control mice. These results reveal that an absence of p75NTR and/or BDNF expression does not perturb the degree of sympathetic innervation of many peripheral tissues during postnatal development, and that a lack of p75NTR expression may actually enhance the density of these efferent fibers in other target tissues, such as the salivary glands. PMID- 15795177 TI - Interaction of central Ang II and NO on the cardiac sympathetic afferent reflex in dogs. AB - The aim of this study was to test the hypothesis that the central angiotensin II (Ang II) and nitric oxide (NO) systems interact to modulate the cardiac sympathetic afferent reflex (CSAR). All dogs were anesthetized with alpha chloralose (100 mg/kg, iv). They were sino-aortic baroreceptor denervated and vagotomized throughout the experiment renal sympathetic nerve activity responses to cardiac sympathetic afferent stimulation and the central gain of the CSAR were measured. Three protocols were performed: (1) intracerebroventricular injection (icv, 3 microg/h or 6 microg/h) of Ang II with and without N(omega)-nitro-L arginine methyl ester (L-NAME) (icv, 1 mg/kg), (2) L-NAME (icv) with and without Ang II (icv, 6 microg/h), and (3) administration of the specific neural NO synthase (nNOS) inhibitor, S-Methyl-L-thiocitrulline (MeTC) (icv, 0.1 or 1 mM, 0.5 ml in 5 min) with and without pretreatment with the angiotensin type 1 receptor antagonist, losartan (icv, 0.125 mg/kg). The primary findings were (1) Ang II alone did not significantly affect the central sensitivity of the CSAR. However, Ang II with L-NAME enhanced this reflex, (2) even though L-NAME alone augmented the CSAR, this excitatory effect was further potentiated in the presence of Ang II and (3) MeTC significantly enhanced the central sensitivity of the CSAR. However, this enhancement did not occur after pretreatment with losartan. These data suggest that Ang II interacts with NO in the brain to modulate the CSAR and that inhibition of NO is required for facilitation of the CSAR by Ang II. PMID- 15795178 TI - Beta-adrenoceptor-mediated vascular relaxation in spontaneously hypertensive rats. AB - Although the impairment of beta-adrenoceptor (beta-AR)-induced vascular relaxation to isoprenaline has been extensively described, discrepancy persisted in the literature. In this work, we investigated beta-AR-induced relaxation in spontaneously hypertensive and normotensive rats aorta. We attempted to determine beta-AR subtypes involved in order to understand the conflicting data regarding the beta-AR-induced vasodilation to isoprenaline. Aortic rings isolated from 12 week-old Wistar Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs) were placed in organ baths and constricted with phenylephrine (alpha1-AR agonist). Then, cumulative concentration-relaxation curves (CCRC) to AR agonists were constructed. In intact aortic rings from both strains, isoprenaline (a nonselective beta-AR agonist) (0.001-10 microM) induced similar concentration dependent relaxations. CCRC was shifted to the right and upward in the presence of nadolol (a nonspecific beta1 and beta2-AR antagonist) (10 microM). After endothelium removal, the response to isoprenaline was partly inhibited in WKY rats, but was strongly inhibited in SHRs. In WKY rats, isoprenaline-induced endothelium-independent relaxation was not modified in the presence of nadolol but was inhibited in the presence of CGP 20712A (low-affinity-state beta1-AR antagonist). In endothelium-denuded rings, SR 58611A (a preferential beta3-AR agonist) (0.1-30 microM) produced a very small relaxation in both strains. In WKY rats, CGP 12177 (CGP) (0.1-30 microM) and cyanopindolol (0.01-3 microM) (partial beta3-AR and low-affinity-state beta1-AR agonists with beta1-AR and beta2-AR antagonistic properties) produced endothelium-independent relaxations. CGP induced effect was significantly inhibited by CGP 20712A (10 microM) or bupranolol (10 microM) (low-affinity-state beta1-AR antagonists). In SHRs, similarly to the impaired endothelium-independent relaxation to isoprenaline, endothelium-independent relaxations to CGP and cyanopindolol were greatly blunted. These relaxations were not modified in the presence of CGP 20712A. In endothelium-denuded rings pretreated with pertussis toxin, CGP-induced relaxation was not modified in WKY rats, but was partly restored in SHRs. In conclusion, these results showed, that in 12-week-old SHRs, the endothelium-independent component of the relaxation to isoprenaline was impaired, and this impairment could involve the low-affinity-state beta1-AR. G(i) protein overexpression and/or overstimulation may be possible factors that contribute to this alteration in hypertension. PMID- 15795179 TI - Origin and pharmacological response of atrial tachyarrhythmias induced by activation of mediastinal nerves in canines. AB - We sought to determine the sites of origin of atrial tachyarrhythmias induced by activating mediastinal nerves, as well as the response of such arrhythmias to autonomic modulation. Under general anaesthesia, atrioventricular block was induced after thoracotomy in 19 canines. Brief trains of 5 electrical stimuli were delivered to right-sided mediastinal nerves during the atrial refractory period. Unipolar electrograms were recorded from 191 right and left atrial epicardial sites under several conditions, i.e. (i) with intact nervous systems and following (ii) acute decentralization of the intrathoracic nervous system or administration of (iii) atropine, (iv) timolol, (v) hexamethonium. Concomitant right atrial endocardial mapping was performed in 7 of these dogs. Mediastinal nerve stimulation consistently initiated bradycardia followed by atrial tachyarrhythmias. In the initial tachyarrhythmia beats, early epicardial breakthroughs were identified in the right atrial free wall (28/50 episodes) or Bachmann bundle region (22/50), which corresponded to endocardial sites of origin associated with the right atrial subsidiary pacemaker complex, i.e. the crista terminalis and dorsal locations including the right atrial aspect of the interatrial septum. Neuronally induced responses were eliminated by atropine, modified by timolol and unaffected by acute neuronal decentralization. After hexamethonium, responses to extra-pericardial but not intra-pericardial nerve stimulation were eliminated. It is concluded that concomitant activation of cholinergic and adrenergic efferent intrinsic cardiac neurons induced by right sided efferent neuronal stimulation initiates atrial tachyarrhythmias that originate from foci anatomically related to the right atrial pacemaker complex and tissues underlying major atrial ganglionated plexuses. PMID- 15795180 TI - Ovarian expression of alpha (1)- and beta (2)-adrenoceptors and p75 neurotrophin receptors in rats with steroid-induced polycystic ovaries. AB - Polycystic ovary syndrome (PCOS) is the main cause of infertility in women. Despite extensive research aimed at identifying the pathogenetic mechanism underlying this condition, the aetiology of the disease is still unknown. Evidence from studies on women with PCOS and on an experimental rat polycystic ovary (PCO) model suggests that the sympathetic regulatory drive to the ovary may be unbalanced. The present study was designed to investigate this hypothesis. Accordingly, we used the well-defined rat PCO model, where PCO is induced by a single intramuscular (i.m.) injection of estradiol valerate (EV), and compared the model with oil-injected controls. We studied the ovarian expression of the alpha1- and beta2-adrenoceptors (ARs), the neurotrophin receptor p75 (p75NTR), and the sympathetic marker tyrosine hydroxylase (TH) at two time points: 30 and 60 days after EV injection. Our data demonstrate for the first time that all of the alpha1-AR subtypes are expressed in normal rat ovaries at both the mRNA and the protein levels. Furthermore, the expression of the alpha1-AR subtypes was differentially modulated in a time- and subtype-dependent manner in rats with EV induced PCO. The ovaries in rats with steroid-induced PCO are characterised by an early overexpression of these molecules and p75NTR, while the beta2-AR was downregulated. An increase in the expression of ovarian TH after EV injection was also detected, suggesting a structural and functional remodelling of ovarian sympathetic innervation in PCO rats. Our evidence strongly indicates that the role of the sympathetic nervous system is crucial in the pathogenesis of EV induced PCO. Overall, our findings suggest that therapeutical approaches aimed at down-regulating the sympathetic tone to the ovary could be useful in the prevention and clinical treatment of PCOS. PMID- 15795181 TI - Excitatory actions of motilin on myenteric neurons of the guinea-pig small intestine. AB - Motilin is considered as a key factor in controlling interdigestive migrating contractions. The present electrophysiological experiments were performed in vitro to examine actions of motilin on myenteric neurons of guinea-pigs after 18 h fasting period. Superfusion of motilin depolarized both S and AH neurons; the lowest effective concentration was 10 nM, and motilin depolarization was observed in 9 of 23 S neurons and in 5 of 25 AH neurons. The motilin depolarizations were associated with an increase in neuronal input resistance. The motilin responses were preserved in Ca2+ free/high Mg2+ solution in which no Ca2+ dependent synaptic transmission occurred. The reversal potential of the motilin responses was estimated about -95 mV, close to the equilibrium potential for K+. Furthermore, muscarinic depolarizations were blocked during the motilin responses. All of these indicated that motilin directly excited myenteric neurons mainly by inactivating K+ channels. It is concluded that motilin might modulate neuronal excitability of the myenteric plexus, leading to the control of interdigestive migrating contractions. PMID- 15795182 TI - Low and high frequency acupuncture stimulation inhibits mental stress-induced sweating in humans via different mechanisms. AB - The effects of acupuncture stimulation at 5 Hz and 100 Hz on mental stress induced sweating were analyzed, and the mechanisms involved were examined using the rate of sweat expulsion as an estimate of central sudomotor outflow. Mental arithmetic was imposed on 25 young healthy volunteers for 2 min before, during and after the stimulation. Acupuncture stimulation was delivered to either the Zusanli (leg) or Hegu (hand) acupoint, and the sweat rate was measured quantitatively during mental arithmetic on the palm or the sole, respectively. When stimulation at 5 Hz was applied to the Zusanli acupoint, the palmar sweat rate (paSR), rate of sweat expulsion (Fsw) and paSR/Fsw were reduced during the stimulation, whereas when it was applied to the Hegu acupoint, plantar SR (plSR) and Fsw were reduced, but plSR/Fsw was not altered. When stimulation at 100 Hz was applied to Zusanli, paSR and paSR/Fsw were reduced, but Fsw was unchanged whereas when it was applied to Hegu, neither plSR, Fsw nor plSR/Fsw was altered. The results suggest that acupuncture stimulation at 5 Hz affects both the supraspinal rhythm-generating mechanism and the mechanisms situated below (probably the spinal cord), whereas stimulation at 100 Hz only affects the mechanisms below the rhythm-generating mechanism. Thus, acupuncture stimulation at 5 Hz and at 100 Hz may reduce mental stress-induced sweating through different mechanisms. PMID- 15795183 TI - Effect of PUVA therapy on electrodermal activity parameters in vitiligo patients. AB - Recent electrophysiological studies on autonomic dysfunction in vitiligo patients show an autonomic dysfunction as measured by absent sympathetic skin response (SSR). Nothing is known about the reversibility of these autonomic parameters with treatment. The present study examined how Electrodermal Activity (EDA) parameters were affected from vitiligo illness before PUVA therapy and whether any electrophysiological gains acquired from PUVA therapy would influence the progression of the condition itself. We studied in 14 healthy subjectS and 14 patients with generalized vitiligo before and after treatment. EDA was recorded with a skin conductance unit connected to a personal computer. Before treatment, skin conductance level (SCL) and habituation number (HN) was higher in vitiligo group than control groups. After treatment, SCL and HN were decreased nearly to normal value. Before and after treatment, prevalence of non-responsivity was higher in vitiligo group than controls. CONCLUSION: The sympathetic nervous system might have a particular role in the pathogenesis of vitiligo. Thus, patients with vitiligo may have changes to EDA parameters that are reversible to great extent with PUVA therapy. EDA is a useful method of studying the autonomic dysfunction in humans. PMID- 15795184 TI - Low-dose atropine amplifies cardiac vagal modulation and increases dynamic baroreflex function in humans. AB - It has been previously known that low-dose atropine (LDA) enhances vagal outflow to the heart. To demonstrate the importance of vagal cardiac modulation in arterial blood pressure (ABP) stability, we evaluated the effect of vagal cardiac stimulation with administration of LDA on ABP fluctuation during dynamic hypertensive and hypotensive stimuli. We assessed changes in RR interval (RRI), ABP, power spectral densities of heart rate variability (HRV) and ABP variability, and spontaneous baroreflex sensitivity (BRS) in 16 healthy volunteers before and after administration of LDA (2 microg/kg). Transient hypertension was induced by phenylephrine (2 microg/kg), whereas hypotension was induced by bilateral thigh cuff deflation after a 3-min suprasystolic occlusion. LDA elicited bradycardia and significantly increased high-frequency (HF, 0.15-0.4 Hz) power of HRV and spontaneous BRS, as determined by transfer function analysis. The increase in systolic blood pressure (SBP) after phenylephrine administration was significantly attenuated by LDA (16+/-2 to 11+/-3 mmHg, P<0.005) and was associated with the augmented reflex bradycardia, whereas the decrease in SBP after cuff deflation was not affected (14+/-5 to 13+/-5 mmHg) with the augmented reflex tachycardia. Increases of HF HRV were correlated significantly and negatively with the increased SBP induced by phenylephrine before and after LDA (r=-0.502, P<0.05). These data suggest that the increased vagal cardiac function induced by LDA augments HR buffering effects, and is important in minimizing arterial pressure fluctuation during dynamic hypertensive stimuli. PMID- 15795185 TI - Sympathetic cardiac influence and arterial blood pressure instability. AB - Previous studies have suggested that sympathetic cardiac blockade enhances baroreflex function, whereas parasympathetic blockade diminishes baroreflex sensitivity and elicits arterial blood pressure (ABP) instability. The aim of this project was to test the hypothesis that sympathetic cardiac blockade was beneficial in maintaining ABP stability during orthostatic challenge. In 8 young healthy subjects, measurements were taken before and after sympathetic cardiac blockade (beta1-adrenoceptor blockade via metoprolol) in combination with or without parasympathetic blockade (atropine) at rest and during lower body negative pressure (LBNP). Arterial blood samples were obtained to evaluate plasma renin activity (PRA) and norepinephrine (NE). Power spectral analyses were performed on heart rate (HR) and ABP variability. LBNP -50 Torr significantly decreased systolic blood pressure (SBP, -6+/-3 mm Hg) and increased PRA (from 0.72+/-0.23 to 1.75+/-0.24 ng ml(-1) h(-1)) and NE (from 1.02+/-0.11 to 2.13+/ 0.32 pg ml(-1)). Low frequency (LF, 0.04-0.12 Hz) SBP and diastolic blood pressure (DBP) variability were significantly augmented by LBNP (4.1+/-1.6 vs. 10.8+/-3.0 mm Hg2, and 3.1+/-1.0 vs. 7.9+/-1.9 mm Hg2, respectively). Following metoprolol, arterial baroreflex sensitivity (assessed by the slope of HR interval to SBP during injection with 1 mug kg(-1) phenylephrine) increased significantly (9.9+/-2.2 to 19.6+/-4.1 ms mm Hg(-1)). With beta1-adrenoceptor blockade, LBNP still decreased SBP (-10+/-2 mm Hg) and increased NE, but did not significantly augment PRA (0.59+/-0.22 vs. 1.03+/-0.18 ng ml(-1) h(-1)), or LF SBP and DBP variability (3.3+/-0.6 vs. 5.7+/-1.3 mm Hg2, and 3.1+/-0.7 vs. 5.4+/-1.1 mm Hg2, respectively). The increased PRA during LBNP remained non-significant following metoprolol combined with atropine, whereas the augmented LF SBP (2.6+/-0.7 vs. 9.9+/-2.8 mm Hg2) and DBP (2.5+/-0.7 vs. 11.1+/-3.0 mm Hg2) variability were significantly accentuated compared to both metoprolol alone and control conditions, accompanied by a greater delta SBP (-17+/-7 mm Hg) and significantly diminished baroreflex gain (0.91+/-0.05 ms/mm Hg). These data suggested that removal of sympathetic cardiac influence improved cardiovascular stability as indicated by a diminished LF ABP variability, which was related to an enhanced cardiac responsiveness. PMID- 15795186 TI - Effects of exercise intensity, posture, pressure on the back and ambient temperature on palmar sweating responses due to handgrip exercises in humans. AB - We have, by using newly developed ratemeters, attempted to examine the effects of exercise intensity, posture, pressure on the skin of the back, and ambient hyperthermic conditions (approximately 30 degrees C) on the 5-s handgrip exercise mediated responses of active palmar sweating in humans. Thirty-five right-handed male (n=5) and female (n=30) volunteer students (20.2+/-1.3 years old) participated in the present study. Oral explanation of only the isometric handgrip exercise (IHG) caused a rapid and oscillatory response (pre-operational) of active palmar sweating in almost all subjects (10 of 14 subjects). Performing the IHG for 5-s caused a significant increase in active sweating rate (operation mediated response) in both ipsi- and contra-lateral palmar surfaces of the thumbs of all subjects. The operation-mediated responses of active palmar sweating to the IHG were reproducible, resulting in no habituation. The increase of operation mediated responses to the IHG was dependent upon exercise intensity (100-25% maximal voluntary contractions). The IHG-mediated ipsi- and contra-lateral responses of active palmar sweating were significantly decreased by changing the body posture from a seated to a supine position or by pressing the skin of the back. Ambient hyperthermic conditions (approximately 30 degrees C) for 60 min also resulted in a significant decrease in the back-pressure-dependent reduction of the operation-mediated responses of active palmar sweating to the IHG. In conclusion, in order to optimize the precision and reproducibility of clinical tests involving palmar sweating responses, it is important that subjects maintain a constant handgrip force and posture and that ambient temperature be kept under normothermic conditions. PMID- 15795187 TI - Autonomic neuropathy may be associated with delayed orocaecal transit time in patients with cirrhosis. AB - Orocaecal transit (OCT) time is delayed in patients with cirrhosis, but the reasons for this remain unclear. We hypothesized that autonomic neuropathy (AN) may explain the delay in OCT. METHODS: We determined OCT and autonomic function tests (AFT) in 48 patients (Child A-15, B-27, C-6) with cirrhosis of various aetiologies. AFT were categorized as normal, borderline, or abnormal. OCT was measured using the lactulose hydrogen (H2) breath test. OCT was defined as the time from baseline when there was a rise in H2 levels of >20 ppm over baseline or >10 ppm over baseline sustained over 2 consecutive time points. RESULTS: Based on OCT, patients were separated into those with delayed OCT (>90 min, group I) and normal OCT (< or = 90 min, group II). Mean OCT time of patients in group I was 169.7+/-49.7 min vs. 84.4+/-12.1 min in group II. Baseline clinical characteristics of patients with and without AN, and those with normal and delayed OCT were similar. Presence of mild encephalopathy did not have an effect on OCT. AN was seen more frequently in group I than group II [16/32 (50%) vs. 3/16 (19%), p=0.03]. Logistic regression analysis showed that the presence of AN was the only independent variable associated with delayed OCT (OR 7.3, CI 1.3 39.4, p=0.02). CONCLUSION: Our study showed that the presence of AN was associated with delayed OCT in patients with cirrhosis. PMID- 15795188 TI - Total elbow arthroplasty: a systematic review of the literature in the English language until the end of 2003. PMID- 15795189 TI - A clinical review of cartilage repair techniques. PMID- 15795190 TI - Mechanical muscle-crush injury and acute muscle-crush compartment syndrome: with special reference to earthquake casualties. PMID- 15795191 TI - Infection in hip arthroplasty after previous injection of steroid. AB - Immunosuppression following intra-articular injections of steroid into the hip may interfere with asepsis in a subsequent total hip arthroplasty (THA). We have undertaken a retrospective, matched, cohort study of infective complications after THA, in 40 patients who had received such an injection and 40 who had not. In the injection group there were five revisions, four of which were for deep infection. There were none in the matched group. The overall rate of revision in our database of 979 primary THAs was 1.02%. Six additional patients who had received injections underwent investigation for infection because of persistent problems in the hip as compared with one in the control group. PMID- 15795192 TI - The Elite Plus total hip arthroplasty: the need for radiological surveillance. AB - We prospectively studied 217 patients who underwent 234 Elite Plus total hip arthroplasties. At a mean of 6.4 (SD 0.7) years post-operatively, 39 patients had died and 22 were either lost to follow-up or had no radiographs available. Clinical (Oxford hip score) and radiological assessments were performed on 156 patients (168 hip arthroplasties) who had a mean age of 67.7 (SD 9.7) years at operation. In the assessed group, 26 of 159 (16.4%) of femoral stems which had not already been revised and 19 of 159 (11.9%) of acetabular cups were definitely loose. In total, 52 of 168 (31%) of hips had either been revised or had definite evidence of loosening of a component. We could not establish any relationship between clinical and radiological outcomes. Despite the fact that the clinical outcome and rate of revision for the Elite Plus appeared to meet international standards, our findings give us cause for concern. We believe that joint registries should include radiological surveillance in order to provide reliable information about medium-term outcomes for hip prostheses. PMID- 15795193 TI - Femoral neck fractures following Birmingham hip resurfacing: a national review of 50 cases. AB - Between April 1999 and April 2004, 3497 Birmingham hips were inserted by 89 surgeons. Fracture of the neck of the femur occurred in 50 patients, an incidence of 1.46%. Complete data were available for 45 patients in whom the incidence was 1.91% for women and 0.98% for men. The relative risk of fracture for women vs men was 1.94961 and this was statistically significant (p < 0.01). The mean time to fracture was 15.4 weeks. In women the fractures occurred at a mean of 18.5 weeks and were more likely to have been preceded by a prodromal phase of pain and limping. In men the mean time to fracture was 13.5 weeks. Significant varus placement of the femoral component, intra-operative notching of the femoral neck and technical problems were common factors in 85% of cases. PMID- 15795194 TI - Radionuclide imaging of the painful hip arthroplasty: positron-emission tomography versus triple-phase bone scanning. AB - Two major complications of hip replacement are loosening and infection. Reliable differentiation between these pathological processes is difficult since both may be accompanied by similar symptoms. Our aim was to assess the diagnostic ability of triple-phase bone scanning (TPBS) and positron-emission tomography (PET) to detect and differentiate these complications in patients with a hip arthroplasty. Both TPBS and PET were performed in 63 patients (92 prostheses). The radiotracer for PET imaging was (18)F-fluorodeoxyglucose (FDG). Image interpretation was performed according to qualitative and quantitative criteria although the final diagnosis was based upon either surgical findings or clinical follow-up. The sensitivity, specificity and accuracy of PET was 0.94, 0.95 and 0.95 respectively, compared with 0.68, 0.76 and 0.74 for TPBS. We found that an image interpretation based exclusively upon quantitative criteria was inappropriate because of its low selectivity. The histological examination indicated that increased periprosthetic uptake of FDG in patients with aseptic loosening was caused by wear-induced polyethylene particles and the subsequent growth of aggressive granulomatous tissue. PMID- 15795195 TI - Joint space width in dysplasia of the hip: a case-control study of 81 adults followed for ten years. AB - In a longitudinal case-control study, we followed 81 subjects with dysplasia of the hip and 136 control subjects without dysplasia for ten years assessing radiological evidence of degeneration of the hip at admission and follow-up. There were no cases of subluxation in the group with dysplasia. Neither subjects with dysplasia nor controls had radiological signs of ongoing degenerative disease at admission. The primary radiological discriminator of degeneration of the hip was a change in the minimum joint space width over time. There were no significant differences between these with dysplasia and controls in regard to age, body mass index or occupational exposure to daily repeated lifting at admission. We found no significant differences in the reduction of the joint space width at follow-up between subjects with dysplasia and the control subjects nor in self-reported pain in the hip. The association of subluxation and/or associated acetabular labral tears with dysplasia of the hip may be a conditional factor for the development of premature osteoarthritis in mildly to moderately dysplastic hips. PMID- 15795196 TI - A hydroxyapatite-coated Insall-Burstein II total knee replacement: 11-year results. AB - We report the clinical and radiographic outcome of a consecutive series of 138 hydroxyapatite-coated total knee replacements with a mean follow-up of 11 years (10 to 13). The patients were entered into a prospective study and all living patients (76 knees) were evaluated. The Hospital for Special Surgery knee score was obtained for comparison with the pre-operative situation. No patient was lost to follow-up. Radiographic assessment revealed no loosening. Seven prostheses have been revised, giving a cumulative survival rate of 93% at 13 years. We believe this to be the longest follow-up report available for an hydroxyapatite coated knee replacement and the first for this design of Insall-Burstein II knee. PMID- 15795197 TI - Contact stress at the post-cam mechanism in posterior-stabilised total knee arthroplasty. AB - We measured the contact areas and contact stresses at the post-cam mechanism of a posterior-stabilised total knee arthroplasty when a posterior force of 500 N was applied to the Kirschner Performance, Scorpio Superflex, NexGen LPS Flex Fixed, and NexGen LPS Flex Mobile knee systems. Measurements were made at 90 degrees, 120 degrees, and 150 degrees of flexion both in neutral rotation and 10 degrees of internal rotation of the tibial component. Peak contact stresses at 90 degrees, 120 degrees, and 150 degrees were 24.0, 33.9, and 28.8 MPa, respectively, for the Kirschner; 26.0, 32.4, and 22.1 MPa, respectively, for the Scorpio; and 34.1, 31.5, and 32.5 MPa, respectively, for the NexGen LPS Flex Fixed. With an internally rotated tibia, the contact stress increased significantly with all the fixed-bearing arthroplasties but not with the NexGen LPS Flex Mobile arthroplasty. The post-cam design should be modified in order to provide a larger contact area whilst avoiding any impingement and edge loading. PMID- 15795198 TI - Ankle fractures in patients with diabetes mellitus. AB - Diabetes mellitus is considered an indicator of poor prognosis for acute ankle fractures, but this risk may be specific to an identifiable subpopulation. We retrospectively reviewed 42 patients with both diabetes mellitus and an acute, closed, rotational ankle fracture. Patients were individually matched to controls by age, gender, fracture type, and surgical vs non-surgical treatment. Outcomes were major complications during the first six months of treatment. We contrasted secondarily 21 diabetic patients with and 21 without diabetic comorbidities. Diabetic patients and controls did not differ significantly in total complication rates. More diabetic patients required long-term bracing. Diabetic patients without comorbidities had complication rates equal to their controls. Diabetic patients with comorbidities had complications at a higher rate (ten patients; 47%) than matched controls (three patients; 14%, p = 0.034). A history of Charcot neuroarthropathy led to the highest rates of complication. An increased risk of complications in diabetic patients with closed rotational fractures of the ankle are specific to a subpopulation with identifiable related comorbidities. PMID- 15795199 TI - Monoblock and modular total shoulder arthroplasty for osteoarthritis. AB - There are theoretical and practical advantages to modular rather than monoblock designs of prostheses for shoulder arthroplasty, but there are no reported studies which specifically compare the clinical and radiological results of their use. We have compared the results of unconstrained total shoulder arthroplasty for osteoarthritis using both types of implant. The monoblock design was used between 1992 and 1995 and the modular design after 1995. Both had cemented all polyethylene glenoids, the monoblock with matched and the modular with mismatched radii of curvature. There were 34 consecutive shoulders in each group with a mean follow-up of 6.1 years in the first and 5.2 years in the second.There were no significant differences in improvement of pain scores, active elevation, external rotation, internal rotation, patient satisfaction, or the Neer ratings between the two groups. Two of 28 glenoid components in the first group and six of 30 in the second met the criteria for being radiologically at risk for loosening (p = 0.25). There were no significant differences in clinical outcome or radiological changes between the first- and second-generation designs of implant for shoulder arthroplasty. PMID- 15795200 TI - Ultrasound-guided needling combined with shock-wave therapy for the treatment of calcifying tendonitis of the shoulder. AB - We carried out a prospective, randomised controlled trial on two groups of 40 patients with painful calcific tendonitis and a mean age of 48.4 years (32.5 to 67.3). All were to undergo arthroscopic removal of the calcific deposit within six months after randomisation. The 40 patients in group I received ultrasound guided needling followed by high-energy shock-wave therapy and the 40 in group II had shock-wave therapy alone. In both groups one treatment consisting of 2500 impulses of shock waves with an energy flux density of 0.36 mJ/mm(2) was applied. The clinical and radiological outcome was assessed using the 100-point Constant shoulder scoring system and standardised radiographs. The mean follow-up was 4.1 months and no patient was lost to follow-up. Both groups had significant improvement in their Constant shoulder score. Radiographs showed disappearance of the calcific deposit in 60.0% of the shoulders in group I and in 32.5% of group II (p < 0.05). Significantly better clinical and radiological results were obtained in group I than in group II. Arthroscopic removal of the deposit was avoided in 32 patients of group I and in 22 of group II. No severe side-effects were recorded.Ultrasound-guided needling in combination with high-energy shock wave therapy is more effective than shock-wave therapy alone in patients with symptomatic calcific tendonitis, giving significantly higher rates of elimination of the calcium deposits, better clinical results and reduction in the need for surgery. PMID- 15795201 TI - The Bryan cervical disc prosthesis as an alternative to arthrodesis in the treatment of cervical spondylosis: 46 consecutive cases. AB - We present data relating to the Bryan disc arthroplasty for the treatment of cervical spondylosis in 46 patients. Patients with either radiculopathy or myelopathy had a cervical discectomy followed by implantation of a cervical disc prosthesis. Patients were reviewed at six weeks, six months and one year and assessment included three outcome measures, a visual analogue scale (VAS), the short form 36 (SF-36) and the neck disability index (NDI). The results were categorised according to a modification of Odom's criteria. Radiological evaluation, by an independent radiologist, sought evidence of movement, stability and subsidence of the prosthesis.A highly significant difference was found for all three outcome measurements, comparing the pre-operative with the post operative values: VAS (Z = 6.42, p < 0.0001), SF-36 (mental component) (Z = 5.02, p < 0.0001), SF-36 (physical component) (Z = -5.00, p < 0.0001) and NDI (Z = 7.03, p < 0.0001). The Bryan cervical disc prosthesis seems reliable and safe in the treatment of patients with cervical spondylosis. PMID- 15795202 TI - Crystal arthropathy of the lumbar spine: a series of six cases and a review of the literature. AB - There have been very few reports in the literature of gout and pseudogout of the spine. We describe six patients who presented with acute sciatica attributable to spinal stenosis with cyst formation in the facet joints. Cytopathological studies confirmed the diagnosis of crystal arthropathy in each case. Specific formation of a synovial cyst was identified pre-operatively by MRI in five patients. In the sixth, the diagnosis was made incidentally during decompressive surgery. Surgical decompression alone was undertaken in four patients. In one with an associated degenerative spondylolisthesis, an additional intertransverse fusion was performed. Another patient had previously undergone a spinal fusion adjacent to the involved spinal segment, and spinal stabilisation was undertaken as well as a decompression. In addition to standard histological examination material was sent for examination under polarised light which revealed deposition of urate or calcium pyrophosphate dihydrate crystals in all cases. It is not possible to diagnose gout and pseudogout of the spine by standard examination of a fixed specimen. However, examining dry specimens under polarised light suggests that crystal arthropathy is a significant aetiological factor in the development of symptomatic spinal stenosis associated with cyst formation in a facet joint. PMID- 15795203 TI - Origin of lumbar spinal roots and their relationship to intervertebral discs: a cadaver and radiological study. AB - Previous studies on the anatomy of the lumbar spine have not clarified the precise relationship of the origin of the lumbar roots to their corresponding discs or their angulation to the dural sac. We studied 33 cadavers (25 formalin preserved and eight fresh-frozen) and their radiographs to determine these details. All cadavers showed a gradual decrease in the angle of the nerve root from L1 to S1. The origin of the root was found to be below the corresponding disc for the L1 to L4 roots. In the formalin-preserved cadavers 8% of the L5 roots originated above, 64% below and 28% at the L4/L5 disc. In the fresh cadavers the values were 12.5%, 62.5% and 25%, respectively. For the S1 root 76% originated above and 24% at the L5-S1 disc in the formalin-preserved cadavers and 75% and 25%, respectively, in the fresh cadavers.A herniated disc usually compresses the root before division of the root sleeve. Thus, compression of the thecal sac before the origin of the root sleeve is common for L1 to L5 whereas compression at the root sleeve is common for S1. Our findings are of value in understanding the pathophysiology of prolapse of the disc and in preventing complications during surgery. PMID- 15795204 TI - Internal fixation versus hemiarthroplasty for displaced fractures of the femoral neck in elderly patients with severe cognitive impairment. AB - We studied 60 patients with an acute displaced fracture of the femoral neck and with a mean age of 84 years. They were randomly allocated to treatment by either internal fixation with cannulated screws or hemiarthroplasty using an uncemented Austin Moore prosthesis. All patients had severe cognitive impairment, but all were able to walk independently before the fracture. They were reviewed at four, 12 and 24 months after surgery. Outcome assessments included complications, revision surgery, the status of activities of daily living (ADL), hip function according to the Charnley score and the health-related quality of life (HRQOL) according to the Euroqol (EQ-5D) (proxy report). General complications and the rate of mortality at two years (42%) did not differ between the groups. The rate of hip complications was 30% in the internal fixation group and 23% in the hemiarthroplasty group; this was not significant. There was a trend towards an increased number of re-operated patients in the internal fixation group compared with the hemiarthroplasty group, 33% and 13%, respectively (p = 0.067), but the total number of surgical procedures which were required did not differ between the groups. Of the survivors at two years, 54% were totally dependent in ADL functions and 60% were bedridden or wheelchair-bound regardless of the surgical procedure. There was a trend towards decreased mobility in the hemiarthroplasty group (p = 0.066). All patients had a very low HRQOL even before the fracture. The EQ-5D(index) score was significantly worse in the hemiarthroplasty group compared with the internal fixation group at the final follow-up (p < 0.001). In our opinion, there is little to recommend hemiarthroplasty with an uncemented Austin Moore prosthesis compared with internal fixation, in patients with severe cognitive dysfunction. PMID- 15795205 TI - Major pelvic fractures: identification of patients at high risk. AB - The identification of high-risk factors in patients with fractures of the pelvis at the time of presentation would facilitate investigation and management. In a series of 174 consecutive patients with unstable fractures of the pelvic ring, clinical data were used to calculate the injury severity score (ISS), the triage revised trauma score (T-RTS), and the Glasgow coma scale (GCS). The morphology of the fracture was classified according to the AO system and that of Burgess et al. The data were analysed using univariate and multivariate methods in order to determine which presenting features were identified with high risk. Univariate analysis showed an association between mortality and an ISS over 25, a T-RTS below eight, age over 65 years, systolic blood pressure under 100 mmHg, a GCS of less than 8, blood transfusion of more than ten units in the first 24 hours and colloid infusion of more than six litres in the first 24 hours. Multivariate analysis showed that age, T-RTS and ISS were independent determinants of mortality. A T-RTS of eight or less identified the cohort of patients at greatest risk (65%). The morphology of the fracture was not predictive of mortality. We recommend the use of the T-RTS in the acute situation in order to identify patients at high risk. PMID- 15795206 TI - Triplate fixation: a new technique in limb-salvage surgery. AB - Massive endoprostheses using a cemented intramedullary stem are widely used to allow early resumption of activity after surgery for tumours. The survival of the prosthesis varies with the anatomical site, the type of prosthesis and the mode of fixation. Revision surgery is required in many cases because of aseptic loosening. Insertion of a second cemented endoprosthesis may be difficult because of the poor quality of the remaining bone, and loosening recurs quickly. We describe a series of 14 patients with triplate fixation in difficult revision or joint-sparing tumour surgery with a minimum follow-up of four years. The triplate design incorporated well within a remodelled cortex to achieve osseomechanical integration with all patients regaining their original level of function within five months. Our preliminary results suggest that this technique may provide an easy, biomechanically friendly alternative to insertion of a further device with an intramedullary stem, which has a shorter lifespan in revision or joint-sparing tumour surgery. A short segment of bone remaining after resection of a tumour will not accept an intramedullary stem, but may be soundly fixed using this method. PMID- 15795207 TI - Ulnar focal cortical indentation: a previously unrecognised form of ulnar dysplasia. AB - Deformity of the forearm due to growth disturbance of the ulna occurs in a number of conditions such as ulnar deficiency, multiple exostoses, and neurofibromatosis. We report a previously unrecognised form, caused by focal cortical indentation. We have treated five children with this condition, three girls and two boys; the mean age at presentation was 5 years (2 to 8). The deformity was first recognised about the age of two years, and progressed gradually. The radiological findings were the same in all cases. The focal cortical indentation was seen at the distal end of the ulna with anteromedial bowing and dysplasia. The radial head was dislocated posterolaterally. In one patient the histological findings at the site of indentation were of a fold of tissue resembling periosteum, which interfered with enchondral ossification. Treatment by ulnar lengthening using an external fixator and osteotomy which corrected both the ulnar deformity and reduced the dislocated radial head in two cases gave the best results. PMID- 15795208 TI - Periacetabular osteotomy in patients with Down's syndrome. AB - We treated eight dysplastic acetabula in six skeletally mature patients with Down's syndrome by a modified Bernese periacetabular osteotomy. The mean age at the time of surgery was 16.5 years (12.8 to 28.5). Mean length of follow-up was five years (2 to 10.4).Pre-operatively the mean (Tonnis) acetabular angle was 28 degrees, the centre-edge angle was -9 degrees, and the extrusion index was 60%; post-operatively they were 3 degrees, 37 degrees, and 17%, respectively. Two patients with post-operative (Tonnis) acetabular angles > 10 degrees developed subluxation post-operatively and required secondary varus derotation femoral osteotomies. Another patient developed a late labral tear which was treated arthroscopically. All eight hips remain clinically stable, and are either asymptomatic or symptomatically improved. These results suggest that the modified Bernese periacetabular osteotomy can be used successfully in the treatment of acetabular dysplasia in patients with Down's syndrome. PMID- 15795209 TI - Multilevel orthopaedic surgery in group IV spastic hemiplegia. AB - Most children with spastic hemiplegia have high levels of function and independence but fixed deformities and gait abnormalities are common. The classification proposed by Winters et al is widely used to interpret hemiplegic gait patterns and plan intervention. However, this classification is based on sagittal kinematics and fails to consider important abnormalities in the transverse plane. Using three-dimensional gait analysis, we studied the incidence of transverse-plane deformity and gait abnormality in 17 children with group IV hemiplegia according to Winters et al before and after multilevel orthopaedic surgery. We found that internal rotation of the hip and pelvic retraction were consistent abnormalities of gait in group-IV hemiplegia. A programme of multilevel surgery resulted in predictable improvement in gait and posture, including pelvic retraction. In group IV hemiplegia pelvic retraction appeared in part to be a compensating mechanism to control foot progression in the presence of medial femoral torsion. Correction of this torsion can improve gait symmetry and function. PMID- 15795210 TI - Glove perforation and contamination in primary total hip arthroplasty. AB - We conducted a randomised, controlled trial to determine whether changing gloves at specified intervals can reduce the incidence of glove perforation and contamination in total hip arthroplasty. A total of 50 patients were included in the study. In the study group (25 patients), gloves were changed at 20-minute intervals or prior to cementation. In the control group (25 patients), gloves were changed prior to cementation. In addition, gloves were changed in both groups whenever there was a visible puncture. Only outer gloves were investigated. Contamination was tested by impression of gloved fingers on blood agar and culture plates were subsequently incubated at 37 degrees C for 48 hours. The number of colonies and types of organisms were recorded. Glove perforation was assessed using the water test. The incidence of perforation and contamination was significantly lower in the study group compared with the control group. Changing gloves at regular intervals is an effective way to decrease the incidence of glove perforation and bacterial contamination during total hip arthroplasty. PMID- 15795211 TI - The management of necrosis-associated and idiopathic bone-marrow oedema of the proximal femur by intravenous iloprost. AB - Bone-marrow oedema can occur both in isolation and in association with necrosis of bone, but it has not been shown whether each respond to the same methods of treatment. We treated 16 patients with isolated oedema and 17, in which it was associated with necrosis of the proximal femur, with the prostacyclin derivative iloprost, which has been shown to be effective in the idiopathic form. The Harris hip score, the range of movement, the extent of the oedema as measured by MRI, pain on a visual analogue scale and patient satisfaction were recorded before and subsequent to treatment. In both groups, we were able to show a significant improvement (p < 0.001) in these observations during the period of follow-up indicating that iloprost will produce clinical improvement in both circumstances. PMID- 15795212 TI - Reduction of high dislocation of the hip using a distraction nail before arthroplasty. AB - We report the case of a 22-year-old woman who underwent plate and screw fixation for a traumatic left acetabular fracture and fixation with cancellous screws for an associated femoral neck fracture. Two months later, the internal fixation became infected and was removed. This resulted in a painful high dislocation of the hip. We solved the problem with continuous soft-tissue distraction using a fully implantable motorised distraction nail in order to reduce the proximal femur prior to total hip arthroplasty. To our knowledge, this is the first time that reduction of a high dislocation of the hip has been performed using such a system. PMID- 15795213 TI - Thoracic penetration following mid-shaft clavicular fracture. AB - We describe a patient who sustained a widely displaced, high-energy, mid-shaft clavicular fracture in association with brachial plexus damage. The distal fragment was subsequently found to have penetrated the thoracic cavity. We describe the treatment of this rare injury with a successful outcome. PMID- 15795214 TI - Systemic cross-linked N-terminal telopeptide and procollagen I C-terminal extension peptide as markers of bone turnover after total hip arthroplasty. AB - There is no diagnostic, non-invasive method for the early detection of loosening after total hip arthroplasty. In a pilot study, we have analysed two serum markers of bone remodelling, procollagen I C-terminal extension peptide (PICP) and cross-linked N-terminal telopeptide (NTx), as well as the diagnostic performance of NTx for the assessment of osteolysis. We recruited 21 patients with loosening (group I), 18 with a well-fixed prosthesis (group II) and 17 at the time of primary arthroplasty for osteoarthritis (OA) (group III). Internal normal reference ranges were obtained from 30 healthy subjects (group IV). The serum PICP level was found to be significantly lower in patients with OA and those with loosening, when compared with those with stable implants, while the NTx level was significantly increased only in the group with loosening, suggesting that collagen degradation depended on the altered bone turnover induced by the implant. This hypothesis was reinforced by the finding that the values in the pre-surgery patients and stable subjects were comparable with the reference range of younger healthy subjects.A high specificity and positive predictive value for NTx provided good diagnostic evidence of agreement between the test and the clinical and radiological evaluations. The NTx level could be used to indicate stability of the implant. However, further prospective, larger studies are necessary. PMID- 15795215 TI - The effects of articular, retinacular, or muscular deficiencies on patellofemoral joint stability: a biomechanical study in vitro. AB - Normal function of the patellofemoral joint is maintained by a complex interaction between soft tissues and articular surfaces. No quantitative data have been found on the relative contributions of these structures to patellar stability. Eight knees were studied using a materials testing machine to displace the patella 10 mm laterally and medially and measure the force required. Patellar stability was tested from 0 degrees to 90 degrees knee flexion with the quadriceps tensed to 175 N. Four conditions were examined: intact, vastus medialis obliquus relaxed, flat lateral condyle, and ruptured medial retinaculae. Abnormal trochlear geometry reduced the lateral stability by 70% at 30 degrees flexion, while relaxation of vastus medialis obliquus caused a 30% reduction. Ruptured medial retinaculae had the largest effect at 0 degrees flexion with 49% reduction. There was no effect on medial stability. There is a complex interaction between these structures, with their contributions to loss of lateral patellar stability varying with knee flexion. PMID- 15795216 TI - Response of the tendon during limb lengthening. AB - Little is known about the increase in length of tendons in postnatal life or of their response to limb lengthening procedures. A study was carried out in ten young and nine adult rabbits in which the tibia was lengthened by 20% at two rates 0.8 mm/day and 1.6 mm/day. The tendon of the flexor digitorum longus (FDL) muscle showed a significant increase in length in response to lengthening of the tibia. The young rabbits exhibited a significantly higher increase in length in the FDL tendon compared with the adults. There was no difference in the amount of lengthening of the FDL tendon at the different rates. Of the increase in length which occurred, 77% was in the proximal half of the tendon. This investigation demonstrated that tendons have the ability to lengthen during limb distraction. This occurred to a greater extent in the young who showed a higher proliferative response, suggesting that there may be less need for formal tendon lengthening in young children. PMID- 15795217 TI - Infection after total hip arthroplasty. PMID- 15795218 TI - Computer-assisted knee arthroplasty versus a conventional jig-based technique. PMID- 15795219 TI - Factors affecting rates of infection and nonunion in intramedullary nailing. PMID- 15795220 TI - Regulation of CAPRICE transcription by MYB proteins for root epidermis differentiation in Arabidopsis. AB - Epidermal cell differentiation in Arabidopsis root is studied as a model system for understanding cell fate specification. Two types of MYB-related transcription factors are involved in this cell differentiation. One of these, CAPRICE (CPC), encoding an R3-type MYB protein, is a positive regulator of hair cell differentiation and is preferentially transcribed in hairless cells. We analyzed the regulatory mechanism of CPC transcription. Deletion analyses of the CPC promoter revealed that hairless cell-specific transcription of the CPC gene required a 69 bp sequence, and a tandem repeat of this region was sufficient for its expression in epidermis. This region includes two MYB-binding sites, and the epidermis-specific transcription of CPC was abolished when base substitutions were introduced in these sites. We showed by gel mobility shift experiments and by yeast one-hybrid assay that WEREWOLF (WER), which is an R2R3-type MYB protein, directly binds to this region. We showed that WER also binds to the GL2 promoter region, indicating that WER directly regulates CPC and GL2 transcription by binding to their promoter regions. PMID- 15795221 TI - The molecular architecture of galactose mutarotase/UDP-galactose 4-epimerase from Saccharomyces cerevisiae. AB - The metabolic pathway by which beta-D-galactose is converted to glucose 1 phosphate is known as the Leloir pathway and consists of four enzymes. In most organisms, these enzymes appear to exist as soluble entities in the cytoplasm. In yeast such as Saccharomyces cerevisiae, however, the first and last enzymes of the pathway, galactose mutarotase and UDP-galactose 4-epimerase, are contained within a single polypeptide chain referred to as Gal10p. Here we report the three dimensional structure of Gal10p in complex with NAD(+), UDP-glucose, and beta-D galactose determined to 1.85-A resolution. The enzyme is dimeric with dimensions of approximately 91 A x 135 A x 108 A and assumes an almost V-shaped appearance. The overall architecture of the individual subunits can be described in terms of two separate N- and C-terminal domains connected by a Type II turn formed by Leu 357 to Val-360. The first 356 residues of Gal10p fold into the classical bilobal topology observed for all other UDP-galactose 4-epimerases studied thus far. This N-terminal domain contains the binding sites for NAD(+) and UDP-glucose. The polypeptide chain extending from Glu-361 to Ser-699 adopts a beta-sandwich motif and harbors the binding site for beta-D-galactose. The two active sites of Gal10p are separated by over 50 A. This investigation represents the first structural analysis of a dual function enzyme in the Leloir pathway. PMID- 15795222 TI - Gating deficiency in a familial hemiplegic migraine type 1 mutant P/Q-type calcium channel. AB - Familial hemiplegic migraine type 1 (FHM1) arises from missense mutations in the gene encoding alpha1A, the pore-forming subunit of P/Q-type calcium channels. The nature of the channel disorder is fundamental to the disease, yet is not well understood. We studied how the most prevalent FHM1 mutation, a threonine to methionine substitution at position 666 (TM), affects both ionic current and gating current associated with channel activation, a previously unexplored feature of P/Q channels. Whole-cell currents were measured in HEK293 cells expressing channels containing either wild-type (WT) or TM alpha1A. Calcium currents were significantly smaller in cells expressing TM channels, consistent with previous reports. In contrast, surface expression of TM channels, measured by immunostaining against an extracellular epitope, was not decreased, and Western blots demonstrated that TM alpha1A subunits were expressed as full-length proteins. WT and TM gating currents were isolated by replacing Ca2+ with the nonpermeant cation La3+. The gating currents generated by the mutant channels were one-third that of WT, a deficiency sufficient to account for the observed attenuation in calcium current; the remaining gating current was no different in kinetics or voltage dependence. Thus, the decreased calcium influx seen with TM channels can be attributed to a reduced number of channels available to undergo the voltage-dependent conformational changes needed for channel opening, not to fewer channel proteins expressed on the cell surface. This identification of an intrinsic defect in FHM1 mutant channels helps explain their impact on neurotransmission when they occupy type-specific slots for P/Q channels at central nerve terminals. PMID- 15795223 TI - Ligand-receptor and receptor-receptor interactions act in concert to activate signaling in the Drosophila toll pathway. AB - In Drosophila, the signaling pathway mediated by the Toll receptor is critical for the establishment of embryonic dorso-ventral pattern and for innate immune responses to bacterial and fungal pathogens. Toll is activated by high affinity binding of the cytokine Spatzle, a dimeric ligand of the cystine knot family. In vertebrates, a related family of Toll-like receptors play a critical role in innate immune responses. Despite the importance of this family of receptors, little is known about the biochemical events that lead to receptor activation and signaling. Here, we show that Spatzle binds to the N-terminal region of Toll and, using biophysical methods, that the binding is complex. The two binding events that cause formation of the cross-linked complex are non-equivalent: the first Toll ectodomain binds Spatzle with an affinity 3-fold higher than the second molecule suggesting that pathway activation involves negative cooperativity. We further show that the Toll ectodomains are able to form low affinity dimers in solution and that juxtamembrane sequences of Toll are critical for the activation or derepression of the pathway. These results, taken together, suggest a mechanism of signal transduction that requires both ligand-receptor and receptor receptor interactions. PMID- 15795224 TI - Myocilin mutations causing glaucoma inhibit the intracellular endoproteolytic cleavage of myocilin between amino acids Arg226 and Ile227. AB - Myocilin is a secreted glycoprotein of unknown function that is ubiquitously expressed in many human organs, including the eye. Mutations in this protein produce glaucoma, a leading cause of blindness worldwide. To explore the biological role of myocilin and the pathogenesis of glaucoma, we have analyzed the expression of recombinant wild type and four representative pathogenic myocilin mutations (E323K, Q368X, P370L, and D380A) in transiently transfected cell lines derived from ocular and nonocular tissues. We found that wild type myocilin undergoes an intracellular endoproteolytic processing at the C terminus of Arg226. This cleavage predicts the production of two fragments, one of 35 kDa containing the C-terminal olfactomedin-like domain, and another of 20 kDa containing the N-terminal leucine zipper-like domain. Here we have analyzed the 35-kDa processed fragment, and we have found that it is co-secreted with the nonprocessed protein. Western immunoblot analyses showed that human aqueous humor and some ocular tissues also contain the processed 35-kDa myocilin, indicating that the endoproteolytic cleavage occurs in vivo. Mutant myocilins accumulated in the endoplasmic reticulum of transfected cells as insoluble aggregates. Interestingly, the four pathogenic myocilins inhibited the endoproteolytic processing with varying efficiency. Furthermore, the mutation P370L, which produces the most severe glaucoma phenotype, also elicited the most potent endoproteolytic cleavage inhibition. We propose that the endoproteolytic processing might regulate the activity of myocilin and that the inhibition of the processing by pathogenic mutations impairs the normal role of myocilin. PMID- 15795225 TI - The serine-rich domain from Crk-associated substrate (p130cas) is a four-helix bundle. AB - p130(cas) (Crk-associated substrate) is a docking protein that is involved in assembly of focal adhesions and concomitant cellular signaling. It plays a role in physiological regulation of cell adhesion, migration, survival, and proliferation, as well as in oncogenic transformation. The molecule consists of multiple protein-protein interaction motifs, including a serine-rich region that is positioned between Crk and Src-binding sites. This study reports the first structure of a functional domain of Cas. The solution structure of the serine rich region has been determined by NMR spectroscopy, demonstrating that this is a stable domain that folds as a four-helix bundle, a protein-interaction motif. The serine-rich region bears strong structural similarity to four-helix bundles found in other adhesion components like focal adhesion kinase, alpha-catenin, or vinculin. Potential sites for phosphorylation and interaction with the 14-3-3 family of cellular regulators are identified in the domain and characterized by site-directed mutagenesis and binding assays. Mapping the degree of amino acid conservation onto the molecular surface reveals a patch of invariant residues near the C terminus of the bundle, which may represent a previously unidentified site for protein interaction. PMID- 15795226 TI - High-throughput screening for potent and selective inhibitors of Plasmodium falciparum dihydroorotate dehydrogenase. AB - Plasmodium falciparum is the causative agent of the most serious and fatal malarial infections, and it has developed resistance to commonly employed chemotherapeutics. The de novo pyrimidine biosynthesis enzymes offer potential as targets for drug design, because, unlike the host, the parasite does not have pyrimidine salvage pathways. Dihydroorotate dehydrogenase (DHODH) is a flavin dependent mitochondrial enzyme that catalyzes the fourth reaction in this essential pathway. Coenzyme Q (CoQ) is utilized as the oxidant. Potent and species-selective inhibitors of malarial DHODH were identified by high-throughput screening of a chemical library, which contained 220,000 drug-like molecules. These novel inhibitors represent a diverse range of chemical scaffolds, including a series of halogenated phenyl benzamide/naphthamides and urea-based compounds containing napthyl or quinolinyl substituents. Inhibitors in these classes with IC(50) values below 600 nm were purified by high pressure liquid chromatography, characterized by mass spectroscopy, and subjected to kinetic analysis against the parasite and human enzymes. The most active compound is a competitive inhibitor of CoQ with an IC(50) against malarial DHODH of 16 nm, and it is 12,500-fold less active against the human enzyme. Site-directed mutagenesis of residues in the CoQ binding site significantly reduced inhibitor potency. The structural basis for the species selective enzyme inhibition is explained by the variable amino acid sequence in this binding site, making DHODH a particularly strong candidate for the development of new anti-malarial compounds. PMID- 15795227 TI - A phosphoethanolamine transferase specific for the outer 3-deoxy-D-manno octulosonic acid residue of Escherichia coli lipopolysaccharide. Identification of the eptB gene and Ca2+ hypersensitivity of an eptB deletion mutant. AB - Addition of a phosphoethanolamine (pEtN) moiety to the outer 3-deoxy-D-manno octulosonic acid (Kdo) residue of lipopolysaccharide (LPS) in WBB06, a heptose deficient Escherichia coli mutant, occurs when cells are grown in 5-50 mM CaCl2 (Kanipes, M. I., Lin, S., Cotter, R. J., and Raetz, C. R. H. (2001) J. Biol. Chem. 276, 1156-1163). A Ca2+-induced, membrane-bound enzyme was responsible for the transfer of the pEtN unit to the Kdo domain. We now report the identification of the gene encoding the pEtN transferase. E. coli yhjW was cloned and overexpressed, because it is homologous to a putative pEtN transferase implicated in the modification of the beta-chain heptose residue of Neisseria meningitidis lipo-oligosaccharide (Mackinnon, F. G., Cox, A. D., Plested, J. S., Tang, C. M., Makepeace, K., Coull, P. A., Wright, J. C., Chalmers, R., Hood, D. W., Richards, J. C., and Moxon, E. R. (2002) Mol. Microbiol. 43, 931-943). In vitro assays with Kdo2-4'-[32P]lipid A as the acceptor showed that YhjW (renamed EptB) utilizes phosphatidylethanolamine in the presence of Ca2+ to transfer the pEtN group. Stoichiometric amounts of diacylglycerol were generated during the EptB-catalyzed transfer of pEtN to Kdo2-lipid A. EptB is an inner membrane protein of 574 amino acid residues with five predicted trans-membrane segments within its N-terminal region. An in-frame replacement of eptB with a kanamycin resistance cassette rendered E. coli WBB06 (but not wild-type W3110) hypersensitive to CaCl2 at 5 mM or higher. Ca2+ hypersensitivity was suppressed by excess Mg2+ in the medium or by restoring the LPS core of WBB06. The latter was achieved by reintroducing the waaC and waaF genes, which encode LPS heptosyl transferases I and II, respectively. Our data demonstrate that pEtN modification of the outer Kdo protected cells containing heptose-deficient LPS from damage by high concentrations of Ca2+. Based on its sequence similarity to EptA(PmrC), we propose that the active site of EptB faces the periplasmic surface of the inner membrane. PMID- 15795228 TI - Substrate specificity and ionic regulation of GlnPQ from Lactococcus lactis. An ATP-binding cassette transporter with four extracytoplasmic substrate-binding domains. AB - We report on the functional characterization of GlnPQ, an ATP-binding cassette transporter with four extracytoplasmic substrate-binding domains. The first predicted transmembrane helix of GlnP was cleaved off in the mature protein and most likely serves as the signal sequence for the extracytoplasmic substrate binding domains. Deletion analysis showed that the substrate-binding domain, in the primary sequence of GlnP nearest to the translocator domain, is used as the receptor that delivers the substrate to the translocator. Membrane reconstitution of the detergent-solubilized and purified GlnPQ complex yielded proteoliposomes that transported glutamine and glutamic acid at the expense of ATP. The transport activity of GlnPQ increased with lumenal salt concentration and internal pH, but the mechanism of ionic activation of the transporter is distinct from that of other osmoregulatory ATP-binding cassette transporters and does not depend on the presence of anionic lipids. The regulation of GlnPQ conforms to an electrostatic switch in which protein domain(s) and low molecular weight electrolytes participate. PMID- 15795229 TI - Mouse Sir2 homolog SIRT6 is a nuclear ADP-ribosyltransferase. AB - Members of the Sir2 family of NAD-dependent protein deacetylases regulate diverse cellular processes including aging, gene silencing, and cellular differentiation. Here, we report that the distant mammalian Sir2 homolog SIRT6 is a broadly expressed, predominantly nuclear protein. Northern analysis of embryonic samples and multiple adult tissues revealed mouse SIRT6 (mSIRT6) mRNA peaks at day E11, persisting into adulthood in all eight tissues examined. At the protein level, mSIRT6 was readily detectable in the same eight tissue types, with the highest levels in muscle, brain, and heart. Subcellular localization studies using both C and N-terminal green fluorescent protein fusion proteins showed mSIRT6-green fluorescent protein to be a predominantly nuclear protein. Indirect immunofluorescence using antibodies to two different mSIRT6 epitopes confirmed that endogenous mSIRT6 is also largely nuclear. Consistent with previous findings, we did not observe any NAD+-dependent protein deacetylase activity in preparations of mSIRT6. However, purified recombinant mSIRT6 did catalyze the robust transfer of radiolabel from [32P]NAD to mSIRT6. Two highly conserved residues within the catalytic core of the protein were required for this reaction. This reaction is most likely mono-ADP-ribosylation because only the modified form of the protein was recognized by an antibody specific to mono-ADP ribose. Surprisingly, we observed that the catalytic mechanism of this reaction is intra-molecular, with individual molecules of mSIRT6 directing their own modification. These results provide the first characterization of a Sir2 protein from phylogenetic class IV. PMID- 15795230 TI - Characterization of a new pantothenate kinase isoform from Helicobacter pylori. AB - Pantothenate kinase (PanK) catalyzes the first step in the biosynthesis of the essential and ubiquitous cofactor coenzyme A (CoA) in all organisms. Two well characterized isoforms of the enzyme are known: a prokaryotic PanK that predominates in eubacteria and a eukaryotic isoform that has primarily been characterized from mammalian and plant sources. Curiously, the genomes of certain pathogenic bacteria, including Helicobacter pylori and Pseudomonas aeruginosa, do not contain a PanK similar to either isoform, although these organisms possess all the other biosynthetic machinery required for CoA production. In this study we cloned, overexpressed and characterized an enzyme from Bacillus subtilis and its homologue from H. pylori and show that they catalyze the ATP-dependent phosphorylation of pantothenate. These enzymes do not share sequence homology with any known PanK, and unlike the bacterial and eukaryotic PanK isoforms their activity is not regulated by either CoA or acetyl-CoA. They also do not accept the pantothenic acid antimetabolite N-pentylpantothenamide as a substrate or are inhibited by it. Taken together, these results point to the identification of a third distinct isoform of PanK that accounts for the only known activity of the enzyme in pathogens such as H. pylori and P. aeruginosa. PMID- 15795231 TI - O-GlcNAcase uses substrate-assisted catalysis: kinetic analysis and development of highly selective mechanism-inspired inhibitors. AB - The post-translational modification of serine and threonine residues of nucleocytoplasmic proteins with 2-acetamido-2-deoxy-d-glucopyranose (GlcNAc) is a reversible process implicated in multiple cellular processes. The enzyme O GlcNAcase catalyzes the cleavage of beta-O-linked GlcNAc (O-GlcNAc) from modified proteins and is a member of the family 84 glycoside hydrolases. The family 20 beta-hexosaminidases bear no apparent sequence similarity yet are functionally related to O-GlcNAcase because both enzymes cleave terminal GlcNAc residues from glycoconjugates. Lysosomal beta-hexosaminidase is known to use substrate-assisted catalysis involving the 2-acetamido group of the substrate; however, the catalytic mechanism of human O-GlcNAcase is unknown. By using a series of 4 methylumbelliferyl 2-deoxy-2-N-fluoroacetyl-beta-D-glucopyranoside substrates, Taft-like linear free energy analyses of these enzymes indicates that O-GlcNAcase uses a catalytic mechanism involving anchimeric assistance. Consistent with this proposal, 1,2-dideoxy-2'-methyl-alpha-D-glucopyranoso-[2,1-d]-Delta2'-thiazoline, an inhibitor that mimics the oxazoline intermediate proposed in the catalytic mechanism of family 20 glycoside hydrolases, is shown to act as a potent competitive inhibitor of both O-GlcNAcase (K(I) = 0.070 microm) and beta hexosaminidase (K = 0.070 microm). A series of 1,2-dideoxy-2'-methyl-alpha-D glucopyranoso-[2,1-d]-Delta2'-thiazoline analogues were prepared, and one inhibitor demonstrated a remarkable 1500-fold selectivity for O-GlcNAcase (K(I) = 0.230 microm) over beta-hexosaminidase (K(I) = 340 microm). These inhibitors are cell permeable and modulate the activity of O-GlcNAcase in tissue culture. Because both enzymes have vital roles in organismal health, these potent and selective inhibitors of O-GlcNAcase should prove useful in studying the role of this enzyme at the organismal level without generating a complex chemical phenotype stemming from concomitant inhibition of beta-hexosaminidase. PMID- 15795232 TI - Antagonistic role of H-NS and GadX in the regulation of the glutamate decarboxylase-dependent acid resistance system in Escherichia coli. AB - One of the most efficient systems of acid resistance in Escherichia coli, the gad system, is based on the coordinated action of two isoforms of glutamate decarboxylase (GadA and GadB) and of a specific glutamate/gamma-aminobutyrate antiporter (GadC). The gadA/BC genes, activated in response to acid stress and in stationary phase cells, are subjected to complex circuits of regulation involving sigma70, sigmaS, cAMP receptor protein, H-NS, EvgAS, TorRS, GadE, GadX, GadW, and YdeO. Herein, we provide evidence that the nucleoid-associated protein H-NS directly functions as repressor of gadA, one of the structural genes, and gadX, a regulatory gene encoding one of the primary activators of the gad system. Band shift and DNase I footprints reveal that H-NS indeed binds to specific sites in the promoter regions of gadA and gadX and represses the transcription of these genes both in an in vitro system and in vivo. Moreover, we show that a maltose binding protein MalE-GadX fusion is able to stimulate the promoter activity of gadA/BC, thus indicating that GadX is by itself able to up-regulate the gad genes and that a functional competition between H-NS and GadX takes place at the gadA promoter. Altogether, our results indicate that H-NS directly inhibits gadA and gadX transcription and, by controlling the intracellular level of the activator GadX, indirectly affects the expression of the whole gad system. PMID- 15795233 TI - Thrombin-induced tyrosine phosphorylation of HS1 in human platelets is sequentially catalyzed by Syk and Lyn tyrosine kinases and associated with the cellular migration of the protein. AB - Thrombin stimulation of platelets triggers Tyr phosphorylation of several signaling proteins, most of which remain unidentified. In this study, we demonstrate for the first time that hematopoietic lineage cell-specific protein 1 (HS1) undergoes a transient Tyr phosphorylation in human platelets stimulated with thrombin. The protein is synergistically phosphorylated by Syk and Lyn tyrosine kinases according to a sequential phosphorylation mechanism. By means of specific inhibitors (PP2, SU6656, and piceatannol) and phosphopeptide-specific antibodies, as well as by coimmunoprecipitation and binding competition experiments, we show that Syk acts as the primary kinase that phosphorylates HS1 at Tyr397 and that Syk phosphorylation is required for HS1 interaction with the Lyn SH2 domain. Upon docking to Syk-phosphorylated HS1, Lyn catalyzes the secondary phosphorylation of the protein at Tyr222. Once the secondary Tyr phosphorylation of HS1 is accomplished the protein dissociates from Lyn and undergoes a dephosphorylation process. HS1 Tyr phosphorylation does not occur when thrombin-induced actin assembly is inhibited by cytochalasin D even under conditions in which Syk and Lyn are still active. Immunofluorescence microscopic analysis shows that the agonist promotes HS1 migration to the plasma membrane and that the inhibition of Lyn-mediated secondary phosphorylation of HS1 abrogates the subcellular translocation of the protein. All together these results indicate that HS1 Tyr phosphorylation catalyzed by Syk and Lyn plays a crucial role in the translocation of the protein to the membrane and is involved in the cytoskeleton rearrangement triggered by thrombin in human platelets. PMID- 15795234 TI - A role for CCR4 in development of mature circulating cutaneous T helper memory cell populations. AB - Expression of the chemokine receptor CCR4 is strongly associated with trafficking of specialized cutaneous memory T helper (Th) lymphocytes to the skin. However, it is unknown whether CCR4 itself participates in the development of cutaneous Th populations. We have addressed this issue via competitive bone marrow (BM) reconstitution assays; equal numbers of BM cells from CCR4(+/+) and CCR4(-/-) donors were allowed to develop side-by-side within RAG-1(-/-) hosts. Cells from both donor types developed equally well into B cells, naive CD8 T cells, naive CD4 T cells, interferon-gamma(+) Th1 cells, and interleukin-4(+) Th2 cells. In marked contrast, circulating cutaneous memory Th cells (i.e., E-selectin ligand(+) [E-lig(+)]) were more than fourfold more likely to be derived from CCR4(+/+) donors than from CCR4(-/-) donors. Most of this effect resides within the CD103(+) subset of the E-lig(+) Th population, in which donor CCR4(+/+) cells can outnumber CCR4(-/-) cells by >12-fold. No similar effect was observed for alpha4beta7(+) intestinal memory Th cells or CD103(+)/E-lig(-) Th cells. We conclude that CCR4 expression provides a competitive advantage to cutaneous Th cells, either by participating in their development from naive Th cells, or by preferentially maintaining them within the memory population over time. PMID- 15795235 TI - Repeated stimulation of CD4 effector T cells can limit their protective function. AB - Chronic infections often result in CD8 T-cell deletion or functional nonresponsiveness. However, to date no definitive studies have attempted to determine the impact of repeated T cell receptor stimulation on CD4 effector T cell generation. We have determined that when antigen presentation is limited to 2 d, optimum in vitro CD4 effector generation is achieved. Alternatively, repeated stimulation results in decreased CD4 effector expansion, decreased cytokine production, and altered migration. Similarly, functionally impaired effectors develop in vivo when antigen-pulsed antigen-presenting cells are replenished every 24 h during a primary immune response. CD4 effectors that are generated with repeated stimulation provide no protection during influenza infection, and have an impaired ability to provide cognate help to B cells. These results suggest that duration of antigen presentation dictates CD4 effector function, and repeated T cell receptor stimulation in vitro and in vivo that exceeds an optimal threshold results in effectors with impaired function. PMID- 15795236 TI - Mutation of the phospholipase C-gamma1-binding site of LAT affects both positive and negative thymocyte selection. AB - Linker for activation of T cells (LAT) is a scaffolding adaptor protein that is critical for T cell development and function. A mutation of LAT (Y136F) that disrupts phospholipase C-gamma1 activation and subsequent calcium influx causes a partial block in T cell development and leads to a severe lymphoproliferative disease in homozygous knock-in mice. One possible contribution to the fatal disease of LAT Y136F knock-in mice could be from autoreactive T cells generated in these mice because of altered thymocyte selection. To examine the impact of the LAT Y136F mutation on thymocyte positive and negative selection, we bred this mutation onto the HY T cell receptor (TCR) transgenic, recombination activating gene-2 knockout background. Female mice with this genotype showed a severe defect in positive selection, whereas male mice exhibited a phenotype resembling positive selection (i.e., development and survival of CD8(hi) HY TCR-specific T cells) instead of negative selection. These results support the hypothesis that in non-TCR transgenic, LAT Y136F knock-in mice, altered thymocyte selection leads to the survival and proliferation of autoreactive T cells that would otherwise be negatively selected in the thymus. PMID- 15795237 TI - Type I interferon dependence of plasmacytoid dendritic cell activation and migration. AB - Differential expression of Toll-like receptor (TLR) by conventional dendritic cells (cDCs) and plasmacytoid DC (pDCs) has been suggested to influence the type of immune response induced by microbial pathogens. In this study we show that, in vivo, cDCs and pDCs are equally activated by TLR4, -7, and -9 ligands. Type I interferon (IFN) was important for pDC activation in vivo in response to all three TLR ligands, whereas cDCs required type I IFN signaling only for TLR9- and partially for TLR7-mediated activation. Although TLR ligands induced in situ migration of spleen cDC into the T cell area, spleen pDCs formed clusters in the marginal zone and in the outer T cell area 6 h after injection of TLR9 and TLR7 ligands, respectively. In vivo treatment with TLR9 ligands decreased pDC ability to migrate ex vivo in response to IFN-induced CXCR3 ligands and increased their response to CCR7 ligands. Unlike cDCs, the migration pattern of pDCs required type I IFN for induction of CXCR3 ligands and responsiveness to CCR7 ligands. These data demonstrate that mouse pDCs differ from cDCs in the in vivo response to TLR ligands, in terms of pattern and type I IFN requirement for activation and migration. PMID- 15795238 TI - Serine protease inhibitors serpina1 and serpina3 are down-regulated in bone marrow during hematopoietic progenitor mobilization. AB - Mobilization of hematopoietic progenitor cells into the blood involves a massive release of neutrophil serine proteases in the bone marrow. We hypothesize that the activity of these neutrophil serine proteases is regulated by the expression of naturally occurring inhibitors (serpina1 and serpina3) produced locally within the bone marrow. We found that serpina1 and serpina3 were transcribed in the bone marrow by many different hematopoietic cell populations and that a strong reduction in expression occurred both at the protein and mRNA levels during mobilization induced by granulocyte colony-stimulating factor or chemotherapy. This decreased expression was restricted to the bone marrow as serpina1 expression was maintained in the liver, leading to no change in plasma concentrations during mobilization. The down-regulation of serpina1 and serpina3 during mobilization may contribute to a shift in the balance between serine proteases and their inhibitors, and an accumulation of active neutrophil serine proteases in bone marrow extravascular fluids that cleave and inactivate molecules essential to the retention of hematopoietic progenitor cells within the bone marrow. These data suggest an unexpected role for serpina1 and serpina3 in regulating the bone marrow hematopoietic microenvironment as well as influencing the migratory behavior of hematopoietic precursors. PMID- 15795239 TI - Protection from cytomegalovirus after transplantation is correlated with immediate early 1-specific CD8 T cells. AB - T cells are crucial for the control of cytomegalovirus (CMV) in infected individuals. Although CMV-specific T cells can be quantified by various methods, clear correlates of protection from CMV disease have not been defined. However, responses to the pp65 protein are believed to play an important role. Here, the proportions of interferon gamma-producing T cells following ex vivo activation with pools of overlapping peptides representing the pp65 and immediate early (IE) 1 proteins were determined at multiple time points and related to the development of CMV disease in 27 heart and lung transplant recipients. Frequencies of IE-1 specific CD8 T cells above 0.2 and 0.4% at day 0 and 2 wk, respectively, or 0.4% at any time during the first months discriminated patients who did not develop CMV disease from patients at risk, 50-60% of whom developed CMV disease. No similar distinction between risk groups was possible based on pp65-specific CD8 or CD4 T cell responses. Remarkably, CMV disease developed exclusively in patients with a dominant pp65-specific CD8 T cell response. In conclusion, high frequencies of IE-1 but not pp65-specific CD8 T cells correlate with protection from CMV disease. These results have important implications for monitoring T cell responses, adoptive cell therapy, and vaccine design. PMID- 15795241 TI - Reflections on a path to sexual commitment. PMID- 15795240 TI - The nuclear factor HMGB1 mediates hepatic injury after murine liver ischemia reperfusion. AB - High-mobility group box 1 (HMGB1) is a nuclear factor that is released extracellularly as a late mediator of lethality in sepsis as well as after necrotic, but not apoptotic, death. Here we demonstrate that in contrast to the delayed role of HMGB1 in the systemic inflammation of sepsis, HMGB1 acts as an early mediator of inflammation and organ damage in hepatic ischemia reperfusion (I/R) injury. HMGB1 levels were increased during liver I/R as early as 1 h after reperfusion and then increased in a time-dependent manner up to 24 h. Inhibition of HMGB1 activity with neutralizing antibody significantly decreased liver damage after I/R, whereas administration of recombinant HMGB1 worsened I/R injury. Treatment with neutralizing antibody was associated with less phosphorylation of c-Jun NH(2)-terminal kinase and higher nuclear factor-kappaB DNA binding in the liver after I/R. Toll-like receptor 4 (TLR4)-defective (C3H/Hej) mice exhibited less damage in the hepatic I/R model than did wild-type (C3H/HeOuj) mice. Anti HMGB1 antibody failed to provide protection in C3H/Hej mice, but successfully reduced damage in C3H/Ouj mice. Together, these results demonstrate that HMGB1 is an early mediator of injury and inflammation in liver I/R and implicates TLR4 as one of the receptors that is involved in the process. PMID- 15795242 TI - Heat shock protein 90 and heat shock protein 70 are components of dengue virus receptor complex in human cells. AB - Dengue virus requires the presence of an unidentified cellular receptor on the surface of the host cell. By using a recently published affinity chromatography approach, an 84-kDa molecule, identified as heat shock protein 90 (HSP90) by matrix-assisted laser desorption ionization-time of flight mass spectrometry, was isolated from neuroblastoma and U937 cells. Based on the ability of HSP90 (84 kDa) to interact with HSP70 (74 kDa) on the surface of monocytes during lipopolysaccharide (LPS) signaling and evidence that LPS inhibits dengue virus infection, the presence of HSP70 was demonstrated in affinity chromatography eluates and by pull-down experiments. Infection inhibition assays support the conclusion that HSP90 and HSP70 participate in dengue virus entry as a receptor complex in human cell lines as well as in monocytes/macrophages. Additionally, our results indicate that both HSPs are associated with membrane microdomains (lipid rafts) in response to dengue virus infection. Moreover, methyl-beta cyclodextrin, a raft-disrupting drug, inhibits dengue virus infection, supporting the idea that cholesterol-rich membrane fractions are important in dengue virus entry. PMID- 15795243 TI - Characterization of homologous and heterologous rotavirus-specific T-cell responses in infant and adult mice. AB - During primary rotavirus (RV) infection, CD8+ T cells play an important role in viral clearance as well as providing partial protection against reinfection. CD4+ T cells are essential for maximal development of RV-specific intestinal immunoglobulin A. In this study, we took advantage of the cytokine flow cytometry technique to obtain a detailed map of H-2b- and H-2d-restricted CD8+ and CD4+ T cell epitopes from the RV proteins VP6 and VP7. Three new CD8+ T-cell epitopes (H 2d and H-2b restricted) and one new CD4+ T-cell epitope (H-2d and H-2b restricted) were identified. Using these newly identified targets, we characterized the development and specificity of cellular immune responses in C57BL/6 and BALB/c mice during acute infection of infants and adults. We found that both the CD4+ and CD8+ responses peaked on days 5 to 7 after infection and then declined rapidly. Interestingly, both the response kinetics and tissue distributions were different when epitopes on VP6 and VP7 were compared. VP6 elicited a response which predominated in the intestine, while the response to VP7 was more systemic. Additionally, the T-cell responses elicited after homologous versus heterologous infection differed substantially. We found that during homologous infection, there was a greater response toward VP6 than that toward VP7, especially in the intestine, while after heterologous infection, this was not the case. Finally, in suckling mice, we found two peaks in the CD8 response on days 7 and 14 postinfection, which differed from the single peak found in adults and likely mimics the biphasic pattern of rotavirus shedding in infant mice. PMID- 15795244 TI - Vaccine-elicited memory cytotoxic T lymphocytes contribute to Mamu-A*01 associated control of simian/human immunodeficiency virus 89.6P replication in rhesus monkeys. AB - The expression of particular major histocompatibility complex (MHC) class I alleles can influence the rate of disease progression following lentiviral infections. This effect is a presumed consequence of potent cytotoxic T lymphocyte (CTL) responses that are restricted by these MHC class I molecules. The present studies have examined the impact of the MHC class I allele Mamu-A*01 on simian/human immunodeficiency virus 89.6P (SHIV-89.6P) infection in unvaccinated and vaccinated rhesus monkeys by exploring the contribution of dominant-epitope specific CTL in this setting. Expression of Mamu-A*01 in immunologically naive monkeys was not associated with improved control of viral replication, CD4+ T-lymphocyte loss, or survival. In contrast, Mamu-A*01+ monkeys that had received heterologous prime/boost immunizations prior to challenge maintained higher CD4+ T-lymphocyte levels and better control of SHIV-89.6P replication than Mamu-A*01- monkeys. This protection was associated with the evolution of high-frequency anamnestic CTL responses specific for a dominant Mamu A*01-restricted Gag epitope following infection. These data indicate that specific MHC class I alleles can confer protection in the setting of a pathogenic SHIV infection by their ability to elicit memory CTL following vaccination. PMID- 15795245 TI - Characterization of DC-SIGN/R interaction with human immunodeficiency virus type 1 gp120 and ICAM molecules favors the receptor's role as an antigen-capturing rather than an adhesion receptor. AB - The dendritic cell (DC)-specific intercellular adhesion molecule 3 (ICAM-3) grabbing nonintegrin binding receptor (DC-SIGN) was shown to bind human immunodeficiency virus type 1 (HIV-1) viral envelope protein gp120 and proposed to function as a Trojan horse to enhance trans-virus infection to host T cells. To better understand the mechanism by which DC-SIGN and DC-SIGNR selectively bind HIV-1 gp120, we constructed a series of deletion mutations in the repeat regions of both receptors. Different truncated receptors exist in different oligomeric forms. The carbohydrate binding domain without any repeats was monomeric, whereas the full extracellular receptors existed as tetramers. All reconstituted receptors retained their ability to bind gp120. The dissociation constant, however, differed drastically from micromolar values for the monomeric receptors to nanomolar values for the tetrameric receptors, suggesting that the repeat region of these receptors contributes to the avidity of gp120 binding. Such oligomerization may provide a mechanism for the receptor to selectively recognize pathogens containing multiple high-mannose-concentration carbohydrates. In contrast, the receptors bound to ICAMs with submicromolar affinities that are similar to those of two nonspecific cell surface glycoproteins, FcgammaRIIb and FcgammaRIII, and the oligomerization of DC-SIGNR resulted in no increase in binding affinity to ICAM-3. These findings suggest that DC-SIGN may not discriminate other cell surface glycoproteins from ICAM-3 binding. The pH dependence in DC-SIGN binding to gp120 showed that the receptor retained high affinity gp120 binding at neutral pH but lost gp120 binding at pH 5, suggesting a release mechanism of HIV in the acidic endosomal compartment by DC-SIGN. Our work contradicts the function of DC-SIGN as a Trojan horse to facilitate HIV-1 infection; rather, it supports the function of DC-SIGN/R (a designation referring to both DC-SIGN and DC-SIGNR) as an antigen-capturing receptor. PMID- 15795246 TI - Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication. AB - RNA elements within flavivirus genomes are potential targets for antiviral therapy. A panel of phosphorodiamidate morpholino oligomers (PMOs), whose sequences are complementary to RNA elements located in the 5'- and 3'-termini of the West Nile (WN) virus genome, were designed to anneal to important cis-acting elements and potentially to inhibit WN infection. A novel Arg-rich peptide was conjugated to each PMO for efficient cellular delivery. These PMOs exhibited various degrees of antiviral activity upon incubation with a WN virus luciferase replicon-containing cell line. Among them, PMOs targeting the 5'-terminal 20 nucleotides (5'End) or targeting the 3'-terminal element involved in a potential genome cyclizing interaction (3'CSI) exhibited the greatest potency. When cells infected with an epidemic strain of WN virus were treated with the 5'End or 3'CSI PMO, virus titers were reduced by approximately 5 to 6 logs at a 5 muM concentration without apparent cytotoxicity. The 3'CSI PMO also inhibited mosquito-borne flaviviruses other than WN virus, and the antiviral potency correlated with the conservation of the targeted 3'CSI sequences of specific viruses. Mode-of-action analyses showed that the 5'End and 3'CSI PMOs suppressed viral infection through two distinct mechanisms. The 5'End PMO inhibited viral translation, whereas the 3'CSI PMO did not significantly affect viral translation but suppressed RNA replication. The results suggest that antisense PMO-mediated blocking of cis-acting elements of flavivirus genomes can potentially be developed into an anti-flavivirus therapy. In addition, we report that although a full-length WN virus containing a luciferase reporter (engineered at the 3' untranslated region of the genome) is not stable, an early passage of this reporting virus can be used to screen for inhibitors against any step of the virus life cycle. PMID- 15795247 TI - The cellular protein daxx interacts with avian sarcoma virus integrase and viral DNA to repress viral transcription. AB - The cellular protein Daxx was identified as an interactor with avian sarcoma virus (ASV) integrase (IN) in a yeast two-hybrid screen. After infection, Daxx-IN interactions were detected by coimmunoprecipitation. An association between Daxx and viral DNA, likely mediated by IN, was also detected by chromatin immunoprecipitation. Daxx was not required for early events in ASV replication, including integration, as Daxx-null cells were transduced as efficiently as Daxx expressing cells. However, viral reporter gene expression from ASV-based vectors was substantially higher in the Daxx-null cells than in Daxx-complemented cells. Consistent with this observation, histone deacetylases (HDACs) were found to associate with viral DNA in Daxx-complemented cells but not in Daxx-null cells. Furthermore, Daxx protein was induced in an interferon-like manner upon ASV infection. We conclude that Daxx interacts with an IN-viral DNA complex early after infection and may mediate the repression of viral gene expression via the recruitment of HDACs. Our findings provide a novel example of cellular immunity against viral replication in which viral transcription is repressed via the recruitment of antiviral proteins to the viral DNA. PMID- 15795248 TI - The Autographa californica multiple nucleopolyhedrovirus ie0-ie1 gene complex is essential for wild-type virus replication, but either IE0 or IE1 can support virus growth. AB - The immediate-early ie0-ie1 gene complex expresses the only baculovirus spliced gene that produces an alternate protein product. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) IE1 is a potent transcriptional transactivator that is essential for viral replication in transient assays. IE1 contains 582 amino acids that are arranged into different domains, including an acidic activation domain at the N terminus, a DNA binding domain, and an oligomerization domain at the C terminus. IE0 is a 52-amino-acid N-terminally elongated form of IE1. We investigated the functions of IE0 and IE1 in virus-infected cells by constructing the first ie1 open reading frame knockout virus. An infectious AcMNPV bacmid was used to generate the ie1 knockout, and the resulting virus, AcBacIE1KO, effectively deletes both ie0 and ie1. AcBacIE1KO does not infect Spodoptera frugiperda cells, showing that the ie0-ie1 gene complex is essential for viral infection. Rescue viruses of AcBacIE1KO were constructed that express only IE1, IE1 and IE0, or only IE0. Our results show that both IE0 and IE1 can function independently, but not equivalently, to support replication, producing infectious virus. Viruses expressing predominately, or only, IE0 produced significantly fewer cells with polyhedra than either the IE1 counterpart or wild-type virus. In addition, DNA replication was prolonged and budded virus and late gene expression were delayed. Viruses expressing only IE1 also produced fewer polyhedra, but replication was slightly faster and achieved higher levels than that of the wild type virus. Both IE0 and IE1 are therefore required and must be expressed in the correct quantitative ratios to achieve a wild-type infection. PMID- 15795249 TI - Requirements at the 3' end of the sindbis virus genome for efficient synthesis of minus-strand RNA. AB - The 3'-untranslated region of the Sindbis virus genome is 0.3 kb in length with a 19-nucleotide conserved sequence element (3' CSE) immediately preceding the 3' poly(A) tail. The 3' CSE and poly(A) tail have been assumed to constitute the core promoter for minus-strand RNA synthesis during genome replication; however, their involvement in this process has not been formally demonstrated. Utilizing both in vitro and in vivo analyses, we have examined the role of these elements in the initiation of minus-strand RNA synthesis. The major findings of this study with regard to efficient minus-strand RNA synthesis are the following: (i) the wild-type 3' CSE and the poly(A) tail are required, (ii) the poly(A) tail must be a minimum of 11 to 12 residues in length and immediately follow the 3' CSE, (iii) deletion or substitution of the 3' 13 nucleotides of the 3' CSE severely inhibits minus-strand RNA synthesis, (iv) templates possessing non-wild-type 3' sequences previously demonstrated to support virus replication do not program efficient RNA synthesis, and (v) insertion of uridylate residues between the poly(A) tail and a non-wild-type 3' sequence can restore promoter function to a limited extent. This study shows that the optimal structure of the 3' component of the minus-strand promoter is the wild-type 3' CSE followed a poly(A) tail of at least 11 residues. Our findings also show that insertion of nontemplated bases can restore function to an inactive promoter. PMID- 15795250 TI - Inhibition of Epstein-Barr virus OriP function by tankyrase, a telomere associated poly-ADP ribose polymerase that binds and modifies EBNA1. AB - Tankyrase (TNKS) is a telomere-associated poly-ADP ribose polymerase (PARP) that has been implicated along with several telomere repeat binding factors in the regulation of Epstein-Barr virus origin of plasmid replication (OriP). We now show that TNKS1 can bind to the family of repeats (FR) and dyad symmetry regions of OriP by using a chromatin immunoprecipitation assay and DNA affinity purification. TNKS1 and TNKS2 bound to EBNA1 in coimmunoprecipitation experiments with transfected cell lysates and with purified recombinant proteins in vitro. Two RXXPDG-like TNKS-interacting motifs in the EBNA1 amino-terminal domain mediated binding with the ankyrin repeat domain of TNKS. Mutations of both motifs at EBNA1 G81 and G425 abrogated TNKS binding and enhanced EBNA1-dependent replication of OriP. Small hairpin RNA targeted knock-down of TNKS1 enhanced OriP dependent DNA replication. Overexpression of TNKS1 or TNKS2 inhibited OriP dependent DNA replication, while a PARP-inactive form of TNKS2 (M1045V) was compromised for this inhibition. We show that EBNA1 is subject to PAR modification in vivo and to TNKS1-mediated PAR modification in vitro. These results indicate that TNKS proteins can interact directly with the EBNA1 protein, associate with the FR region of OriP in vivo, and inhibit OriP replication in a PARP-dependent manner. PMID- 15795251 TI - Activation of CD21 and CD23 gene expression by Kaposi's sarcoma-associated herpesvirus RTA. AB - Epstein-Barr virus (EBV) EBNA2 and Kaposi's sarcoma-associated herpesvirus (KSHV) replication and transcription activator (RTA) are recruited to their responsive elements through interaction with a Notch-mediated transcription factor, RBP Jkappa. In particular, RTA and EBNA2 interactions with RBP-Jkappa are essential for the lytic replication of KSHV and expression of B-cell activation markers CD21 and CD23a, respectively. Here, we demonstrate that like EBV EBNA2, KSHV RTA strongly induces CD21 and CD23a expression through RBP-Jkappa binding sites in the first intron of CD21 and in the CD23a core promoter, respectively. However, unlike EBV EBNA2, which alters immunoglobulin mu (Igmu) and c-myc gene expression, RTA did not affect Igmu and c-myc expression, indicating that KSHV RTA targets the Notch signal transduction pathway in a manner similar to but distinct from that of EBV EBNA2. Furthermore, RTA-induced expression of CD21 glycoprotein, which is an EBV receptor, efficiently facilitated EBV infection. In addition, RTA-induced CD23 glycoprotein underwent proteolysis and gave rise to soluble CD23 (sCD23) molecules in B lymphocytes and KSHV-infected primary effusion lymphocytes. sCD23 then stimulated primary human lymphocytes. These results demonstrate that cellular CD21 and CD23a are common targets for B lymphotropic gammaherpesviruses and that KSHV RTA regulates RBP-Jkappa-mediated cellular gene expression, which ultimately provides a favorable milieu for viral reproduction in the infected host. PMID- 15795253 TI - Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: fine mapping and escape mutant analysis. AB - Anti-rabies virus immunoglobulin combined with rabies vaccine protects humans from lethal rabies infections. For cost and safety reasons, replacement of the human or equine polyclonal immunoglobulin is advocated, and the use of rabies virus-specific monoclonal antibodies (MAbs) is recommended. We produced two previously described potent rabies virus-neutralizing human MAbs, CR57 and CRJB, in human PER.C6 cells. The two MAbs competed for binding to rabies virus glycoprotein. Using CR57 and a set of 15-mer overlapping peptides covering the glycoprotein ectodomain, a neutralization domain was identified between amino acids (aa) 218 and 240. The minimal binding region was identified as KLCGVL (aa 226 to 231), with key residues K-CGV- identified by alanine replacement scanning. The critical binding region of this novel nonconformational rabies virus epitope is highly conserved within rabies viruses of genotype 1. Subsequently, we generated six rabies virus variants escaping neutralization by CR57 and six variants escaping CRJB. The CR57 escape mutants were only partially covered by CRJB, and all CRJB-resistant variants completely escaped neutralization by CR57. Without exception, the CR57-resistant variants showed a mutation at key residues within the defined minimal binding region, while the CRJB escape viruses showed a single mutation distant from the CR57 epitope (N182D) combined with mutations in the CR57 epitope. The competition between CR57 and CRJB, the in vitro escape profile, and the apparent overlap between the recognized epitopes argues against including both CR57 and CRJB in a MAb cocktail aimed at replacing classical immunoglobulin preparations. PMID- 15795252 TI - Precise identification of endogenous proviruses of NFS/N mice participating in recombination with moloney ecotropic murine leukemia virus (MuLV) to generate polytropic MuLVs. AB - Polytropic murine leukemia viruses (MuLVs) are generated by recombination of ecotropic MuLVs with env genes of a family of endogenous proviruses in mice, resulting in viruses with an expanded host range and greater virulence. Inbred mouse strains contain numerous endogenous proviruses that are potential donors of the env gene sequences of polytropic MuLVs; however, the precise identification of those proviruses that participate in recombination has been elusive. Three different structural groups of proviruses in NFS/N mice have been described and different ecotropic MuLVs preferentially recombine with different groups of proviruses. In contrast to other ecotropic MuLVs such as Friend MuLV or Akv that recombine predominantly with a single group of proviruses, Moloney MuLV (M-MuLV) recombines with at least two distinct groups. In this study, we determined that only three endogenous proviruses, two of one group and one of another group, are major participants in recombination with M-MuLV. Furthermore, the distinction between the polytropic MuLVs generated by M-MuLV and other ecotropic MuLVs is the result of recombination with a single endogenous provirus. This provirus exhibits a frameshift mutation in the 3' region of the surface glycoprotein-encoding sequences that is excluded in recombinants with M-MuLV. The sites of recombination between the env genes of M-MuLV and endogenous proviruses were confined to a short region exhibiting maximum homology between the ecotropic and polytropic env sequences and maximum stability of predicted RNA secondary structure. These observations suggest a possible mechanism for the specificity of recombination observed for different ecotropic MuLVs. PMID- 15795255 TI - A substitution in rous sarcoma virus integrase that separates its two biologically relevant enzymatic activities. AB - Retroviral integrase prepares viral DNA for integration by removing 2 nucleotides from each end of unintegrated DNA in a reaction referred to as processing. However, it has been known since the processing assay was first described that avian integrases frequently nick 3 nucleotides, as well as 2 nucleotides, from viral DNA ends when reaction mixtures contain Mn2+. We now report that specificity for the biologically relevant "-2" site is enhanced when the serine at amino acid 124 of Rous sarcoma virus (RSV) integrase is replaced by alanine, valine, glycine, lysine, or aspartate. The protein with a serine-to-aspartate substitution exhibited especially high fidelity for the correct site, as evidenced by a ratio of -2 nicks to -3 nicks that was more than 40-fold greater than that for the wild-type enzyme in reactions with Mn2+. Even with Mg2+, the substituted proteins exhibited greater specificity than the wild type, especially the S124D protein. Moreover, this protein was more efficient than the wild type at processing viral DNA ends. Unexpectedly, however, the S124D protein was significantly impaired at catalyzing the insertion of viral DNA ends in reactions with Mn2+ and joining was undetectable in reactions with Mg2+. Thus, the S124D protein has separated the processing and joining activities of integrase. Similar results were found for human immunodeficiency virus integrase with the analogous substitution. No proteins with comparable properties have been described. Moreover, RSV virions containing integrase with the S124D mutation were unable to replicate in cell cultures. Together, these data suggest that integrase has evolved to have submaximal processing activity so that it can also catalyze DNA joining. PMID- 15795254 TI - Expression of CD154 by a simian immunodeficiency virus vector induces only transitory changes in rhesus macaques. AB - Human immunodeficiency virus infection is characterized by dysregulation of antigen-presenting cell function and defects in cell-mediated immunity. Recent evidence suggests that impaired ability of CD4+ T cells to upregulate the costimulatory molecule CD154 is at the core of this dysregulation. To test the hypothesis that increased expression of CD154 on infected CD4+ T cells could modulate immune function, we constructed a replication-competent simian immunodeficiency virus (SIV) vector that expressed CD154. We found that this recombinant vector directed the expression of CD154 on the surface of infected CD4+ T cells and that expression of CD154 resulted in activation of B cells present in the same cultures. Experimental infection of rhesus macaques resulted in very low viral loads for the CD154-expressing virus and the control virus, indicating that expression of CD154 did not result in increased viral replication. Analyses of the anti-SIV immune responses and the phenotype of lymphocytes in blood and lymphoid tissues showed changes that occurred during the acute phase of infection only in animals infected with the CD154-expressing SIV, but that became indistinguishable from those seen in animals infected with the control virus at later time points. We conclude that the level of expression of CD154 in itself is not responsible for affecting the immune response to an attenuated virus. Considering that the CD154-expressing SIV vector and the virus control did not carry an active nef gene, our results suggest that, in CD4+ T cells infected with wild-type virus, Nef is the viral factor that interferes with the immune mechanisms that regulate expression of CD154. PMID- 15795256 TI - Virus-specific and bystander CD8 T cells recruited during virus-induced encephalomyelitis. AB - Neurotropic coronavirus-induced encephalitis was used to evaluate recruitment, functional activation, and retention of peripheral bystander memory CD8+ T cells. Mice were first infected with recombinant vaccinia virus expressing a non-cross reactive human immunodeficiency virus (HIV) epitope, designated p18. Following establishment of an endogenous p18-specific memory CD8+ T-cell population, mice were challenged with coronavirus to directly compare recruitment, longevity, and activation characteristics of both primary coronavirus-specific and bystander memory populations trafficking into the central nervous system (CNS). HIV specific memory CD8+ T cells were recruited early into the CNS as components of the innate immune response, preceding CD8+ T cells specific for the dominant coronavirus epitope, designated pN. Although pN-specific T-cell numbers gradually exceeded bystander p18-specific CD8+ T-cell numbers, both populations peaked concurrently within the CNS. Nevertheless, coronavirus-specific CD8+ T cells were preferentially retained. By contrast, bystander CD8+ T-cell numbers declined to background numbers following control of CNS virus replication. Furthermore, in contrast to highly activated pN-specific CD8+ T cells, bystander p18-specific CD8+ T cells recruited to the site of inflammation maintained a nonactivated memory phenotype and did not express ex vivo cytolytic activity. Therefore, analysis of host CD8+ T-cell responses to unrelated infections demonstrates that bystander memory CD8+ T cells can comprise a significant proportion of CNS inflammatory cells during virus-induced encephalitis. However, transient CNS retention and the absence of activation suggest that memory bystander CD8+ T cells may not overtly contribute to pathology in the absence of antigen recognition. PMID- 15795257 TI - Association of ebola virus matrix protein VP40 with microtubules. AB - Viruses exploit a variety of cellular components to complete their life cycles, and it has become increasingly clear that use of host cell microtubules is a vital part of the infection process for many viruses. A variety of viral proteins have been identified that interact with microtubules, either directly or via a microtubule-associated motor protein. Here, we report that Ebola virus associates with microtubules via the matrix protein VP40. When transfected into mammalian cells, a fraction of VP40 colocalized with microtubule bundles and VP40 coimmunoprecipitated with tubulin. The degree of colocalization and microtubule bundling in cells was markedly intensified by truncation of the C terminus to a length of 317 amino acids. Further truncation to 308 or fewer amino acids abolished the association with microtubules. Both the full-length and the 317 amino-acid truncation mutant stabilized microtubules against depolymerization with nocodazole. Direct physical interaction between purified VP40 and tubulin proteins was demonstrated in vitro. A region of moderate homology to the tubulin binding motif of the microtubule-associated protein MAP2 was identified in VP40. Deleting this region resulted in loss of microtubule stabilization against drug induced depolymerization. The presence of VP40-associated microtubules in cells continuously treated with nocodazole suggested that VP40 promotes tubulin polymerization. Using an in vitro polymerization assay, we demonstrated that VP40 directly enhances tubulin polymerization without any cellular mediators. These results suggest that microtubules may play an important role in the Ebola virus life cycle and potentially provide a novel target for therapeutic intervention against this highly pathogenic virus. PMID- 15795258 TI - Role of the specific amino acid sequence of the membrane-spanning domain of human immunodeficiency virus type 1 in membrane fusion. AB - Fusion between cell and virus membranes mediated by gp41 initiates the life cycle of human immunodeficiency virus type 1. In contrast to the many studies that have elucidated the structure-function relationship of the ectodomain, the study of the membrane-spanning domain (MSD) has been rather limited. In particular, the role that the MSD's specific amino acid sequences may have in membrane fusion as well as other gp41 functions is not well understood. The MSD of gp41 contains well-conserved glycine residues that form the GXXXG motif (G, glycine; X, other amino acid residues), a motif often found at the helix-helix interface of membrane spanning alpha-helices. Here we examined the role that the specific amino acid sequence of the gp41 MSD has in gp41 function, particularly in membrane fusion, by making two types of MSD mutants: (i) glycine substitution mutants in which glycine residues of the MSD were mutated to alanine or leucine residues, and (ii) replacement mutants in which the entire MSD was replaced with one derived from glycophorin A or from vesicular stomatitis virus G. The substitution of glycines did not affect gp41 function. MSD-replacement mutants, however, showed severely impaired fusion activity. The assay using the Env expression vector revealed defects in membrane fusion after CD4 binding steps in the MSD-replacement mutants. In addition, the change in Env processing was noted for MSD-replacement mutants. These results suggest that the MSD of gp41 has a relatively wide but not unlimited tolerance for mutations and plays a critical role in membrane fusion as well as in other steps of Env biogenesis. PMID- 15795260 TI - Vaccinia virus H2 protein is an essential component of a complex involved in virus entry and cell-cell fusion. AB - The vaccinia virus H2R gene (VACWR 100) is conserved in all sequenced members of the poxvirus family and encodes a protein with a predicted transmembrane domain and four invariant cysteines. A recombinant vaccinia virus, in which expression of the H2 protein is stringently regulated, was unable to replicate without inducer. However, under nonpermissive conditions, all stages of virus morphogenesis appeared normal and extracellular virions were detected at the tips of actin tails. Nevertheless, virus did not spread to neighboring cells nor did syncytia form after low-pH treatment. Purified -H2 and +H2 virions from cells infected in the absence or presence of inducer, respectively, were indistinguishable in microscopic appearance and contained the same complement of major proteins, though only +H2 virions were infectious. The -H2 virions bound to cells, but their cores did not penetrate into the cytoplasm. In addition, exogenously added -H2 virions were unable to mediate the formation of syncytia after low-pH treatment. In contrast, virions lacking the A27 (p14) protein, which was previously considered to have an essential role in fusion, penetrated cells and induced extensive syncytia. The properties of H2, however, are very similar to those recently reported for the A28 protein. Moreover, coimmunoprecipitation experiments indicated an interaction between H2 and A28. Therefore, H2 and A28 are the only proteins presently known to be specifically required for vaccinia virus entry and are likely components of a fusion complex. PMID- 15795259 TI - Nuclear localizations of the herpes simplex virus type 1 tegument proteins VP13/14, vhs, and VP16 precede VP22-dependent microtubule reorganization and VP22 nuclear import. AB - Herpes simplex virus type 1 (HSV-1) induces microtubule reorganization beginning at approximately 9 h postinfection (hpi), and this correlates with the nuclear localization of the tegument protein VP22. Thus, the active retention of this major virion component by cytoskeletal structures may function to regulate its subcellular localization (A. Kotsakis, L. E. Pomeranz, A. Blouin, and J. A. Blaho, J. Virol. 75:8697-8711, 2001). The goal of this study was to determine whether the subcellular localization patterns of other HSV-1 tegument proteins are similar to that observed with VP22. To address this, we performed a series of indirect immunofluorescence analyses using synchronously infected cells. We observed that tegument proteins VP13/14, vhs, and VP16 localized to the nucleus as early as 5 hpi and were concentrated in nuclei by 9 hpi, which differed from that seen with VP22. Microtubule reorganization was delayed during infection with HSV-1(RF177), a recombinant virus that does not produce full-length VP22. These infected cells did not begin to lose microtubule-organizing centers until 13 hpi. Repair of the unique long 49 (UL49) locus in HSV-1(RF177) yielded HSV-1(RF177R). Microtubule reorganization in HSV-1(RF177R)-infected cells occurred with the same kinetics as HSV-1(F). Acetylated tubulin remained unchanged during infection with either HSV-1(F) or HSV-1(RF177). Thus, while alpha-tubulin reorganized during infection, acetylated tubulin was stable, and the absence of full-length VP22 did not affect this stability. Our findings indicate that the nuclear localizations of tegument proteins VP13/14, VP16, and vhs do not appear to require HSV-1 induced microtubule reorganization. We conclude that full-length VP22 is needed for optimal microtubule reorganization during infection. This implies that VP22 mainly functions to reorganize microtubules later, rather than earlier, in infection. That acetylated tubulin does not undergo restructuring during VP22 dependent, virus-induced microtubule reorganization suggests that it plays a role in stabilizing the infected cells. Our results emphasize that VP22 likely plays a key role in cellular cytopathology during HSV-1 infection. PMID- 15795261 TI - Visualization and characterization of the intracellular movement of vaccinia virus intracellular mature virions. AB - Previous work indicated that vaccinia intracellular mature virus (IMV) utilizes microtubules to move from the viral factory to the site of intracellular envelopment and that expression of the viral A27 protein is required for this transport. To investigate further the role of A27 in IMV intracellular transport, a recombinant vaccinia virus was constructed that had the A27L gene deleted and expressed a yellow fluorescent protein (YFP)-A4 chimera in place of the normal A4 protein. The resulting recombinant, vYFP-A4/DeltaA27, produced relatively normal quantities of virus in a one-step growth curve but had a small plaque phenotype. Subsequent experiments demonstrated that vYFP-A4/DeltaA27 was severely defective in envelope virus production. Despite the absence of A27, live digital video fluorescent microscopy visualized YFP-labeled IMV movement in cells infected with the recombinant. Virion movement approached 3 mum/s and was sensitive to the microtubule depolymerizing drug nocodazole. In addition, IMV could be discerned transiting away from and back towards viral factories. Immunofluorescent staining determined that the distance traveled by A27-deficient virions was sufficient for transport to the site of envelopment. These results indicate that IMVs are capable of bidirectional movement on microtubules, suggesting that they are able to interact with both kinesin and dynein microtubule motors in the absence of A27 and that the distance traveled is sufficient to deliver IMV to the site of wrapping. PMID- 15795262 TI - Expression of matrix metalloproteinases and their tissue inhibitor during viral encephalitis. AB - Matrix metalloproteinases (MMPs) participate in remodeling the extracellular matrix and facilitate entry of inflammatory cells into tissues. Infection of the murine central nervous system (CNS) with a neurotropic coronavirus induces encephalitis associated with increased levels of mRNA encoding MMP-3 and MMP-12. Whereas virus-induced MMP-3 expression was restricted to CNS resident astrocytes, MMP-12 mRNA was expressed by both inflammatory cells and CNS resident cells. Immunosuppression increased both MMP-3 and MMP-12 mRNA levels in CNS resident cells, suggesting that the presence of virus rather than inflammation induced protease up-regulation. MMP activity is partially regulated by a small family of genes encoding tissue inhibitors of matrix metalloproteinases (TIMPs); among the TIMPs, only TIMP-1 mRNA expression increased in the CNS following coronavirus infection. During inflammation TIMP-1 mRNA was most prominently expressed by infiltrating cells. By contrast, in the immunosuppressed host TIMP-1 mRNA was expressed by CNS resident cells. Analysis of cytokine and chemokine mRNA induction within the infected CNS of healthy and immunocompromised mice suggested a possible correlation between increased viral replication and increased levels of beta interferon, MMP-3, MMP-12, and TIMP-1 mRNA. CD4+ T cells which localize to the perivascular and subarachnoid spaces were identified as the primary source of TIMP-1 protein. By contrast, protein expression was undetectable in astrocytes or CD8+ T cells, the primary antiviral effectors that localize to the CNS parenchyma in response to infection. These data suggest that in contrast to the results seen with MMPs, inhibition of protease activity via TIMP-1 expression correlates with the differential tissue distribution of T-cell subsets during acute coronavirus-induced encephalitis. PMID- 15795263 TI - Human immunodeficiency virus (HIV) gp41 escape mutants: cross-resistance to peptide inhibitors of HIV fusion and altered receptor activation of gp120. AB - Human immunodeficiency virus (HIV) infects cells by fusing with cellular membranes. Fusion occurs when the envelope glycoprotein (Env) undergoes conformational changes while binding to cellular receptors. Fusogenic changes involve assembly of two heptad repeats in the ectodomain of the gp41 transmembrane subunit to form a six-helix bundle (6HB), consisting of a trimeric N heptad repeat (N-HR) coiled-coil core with three antiparallel C heptad repeats (C-HRs) that pack in the coiled-coil grooves. Peptides corresponding to the N-and C-HRs (N and C peptides, respectively) interfere with formation of the 6HB in a dominant-negative manner and are emerging as a new class of antiretroviral therapeutics for treating HIV infection. We generated an escape mutant virus with resistance to an N peptide and show that early resistance involved two mutations, one each in the N- and C-HRs. The mutations conferred resistance not only to the selecting N peptide but also to C peptides, as well as other types of N-peptide inhibitors. Moreover, the N-HR mutation altered sensitivity to soluble CD4. Biophysical studies suggest that the 6HB with the resistance mutations is more stable than the wild-type 6HB and the 6HB formed by inhibitor binding to either wild-type or mutant C-HR. These findings provide new insights into potential mechanisms of resistance to HIV peptide fusion inhibitors and dominant-negative inhibitors in general. The results are discussed in the context of current models of Env-mediated membrane fusion. PMID- 15795264 TI - Inhibition of murine leukemia virus envelope protein (env) processing by intracellular expression of the env N-terminal heptad repeat region. AB - A conserved structural motif in the envelope proteins of several viruses consists of an N-terminal, alpha-helical, trimerization domain and a C-terminal region that refolds during fusion to bind the N-helix trimer. Interaction between the N and C regions is believed to pull viral and target membranes together in a crucial step during membrane fusion. For several viruses with type I fusion proteins, C regions pack as alpha-helices in the grooves between N-helix monomers, and exogenously added N- and C-region peptides block fusion by inhibiting the formation of the six-helix bundle. For other viruses, including influenza virus and murine leukemia virus (MLV), there is no evidence for comparably extended C-region alpha-helices, although a short, non-alpha-helical interaction structure has been reported for influenza virus. We tested candidate N-helix and C-region peptides from MLV for their ability to inhibit cell fusion but found no inhibitory activity. In contrast, intracellular expression of the MLV N-helix inhibited fusion by efficiently blocking proteolytic processing and intracellular transport of the envelope protein. The results highlight another mechanism by which the N-helix peptides can inhibit fusion. PMID- 15795265 TI - Comprehensive analysis of ebola virus GP1 in viral entry. AB - Ebola virus infection is initiated by interactions between the viral glycoprotein GP1 and its cognate receptor(s), but little is known about the structure and function of GP1 in viral entry, partly due to the concern about safety when working with the live Ebola virus and the difficulty of manipulating the RNA genome of Ebola virus. In this study, we have used a human immunodeficiency virus based pseudotyped virus as a surrogate system to dissect the role of Ebola virus GP1 in viral entry. Analysis of more than 100 deletion and amino acid substitution mutants of GP1 with respect to protein expression, processing, viral incorporation, and viral entry has allowed us to map the region of GP1 responsible for viral entry to the N-terminal 150 residues. Furthermore, six amino acids in this region have been identified as critical residues for early events in Ebola virus entry, and among these, three are clustered and are implicated as part of a potential receptor-binding pocket. In addition, substitutions of some 30 residues in GP1 are shown to adversely affect GP1 expression, processing, and viral incorporation, suggesting that these residues are involved in the proper folding and/or overall conformation of GP. Sequence comparison of the GP1 proteins suggests that the majority of the critical residues for GP folding and viral entry identified in Ebola virus GP1 are conserved in Marburg virus. These results provide information for elucidating the structural and functional roles of the filoviral glycoproteins and for developing potential therapeutics to block viral entry. PMID- 15795266 TI - The mitotic chromosome binding activity of the papillomavirus E2 protein correlates with interaction with the cellular chromosomal protein, Brd4. AB - The papillomavirus transcriptional activator, E2, is involved in key functions of the viral life cycle. These include transcriptional regulation, viral DNA replication, and viral genome segregation. The transactivation domain of E2 is required for each of these functions. To identify the regions of the domain that mediate binding to mitotic chromosomes, a panel of mutations has been generated and their effect on various E2 functions has been analyzed. A structural model of the bovine papillomavirus type 1 (BPV1) E2 transactivation domain was generated based on its homology with the solved structure of the human papillomavirus type 16 (HPV16) domain. This model was used to identify distinct surfaces of the domain to be targeted by point mutation to further delineate the functional region of the transactivation domain responsible for mitotic chromosome association. The mutated E2 proteins were assessed for mitotic chromosome binding and, in addition, transcriptional activation and transcriptional repression activities. Mutation of amino acids R37 and I73, which are located on a surface of the domain that in HPV16 E2 is reported to mediate self-interaction, completely eliminated mitotic chromosome binding. Mitotic chromosome binding activity was found to correlate well with the ability to interact with the cellular chromosomal associated factor Brd4, which has recently been proposed to mediate the association between BPV1 E2 and mitotic chromosomes. PMID- 15795267 TI - Role of the varicella-zoster virus gene product encoded by open reading frame 35 in viral replication in vitro and in differentiated human skin and T cells in vivo. AB - Although genes related to varicella-zoster virus (VZV) open reading frame 35 (ORF35) are conserved in the herpesviruses, information about their contributions to viral replication and pathogenesis is limited. Using a VZV cosmid system, we deleted ORF35 to produce two null mutants, designated rOkaDelta35(#1) and rOkaDelta35(#2), and replaced ORF35 at a nonnative site, generating two rOkaDelta35/35@Avr mutants. ORF35 Flag-tagged recombinants were made by inserting ORF35-Flag at the nonnative Avr site as the only copy of ORF35, yielding rOkaDelta35/35Flag@Avr, or as a second copy, yielding rOka35Flag@Avr. Replication of rOkaDelta35 viruses was diminished in melanoma and Vero cells in a 6-day analysis of growth kinetics. Plaque sizes of rOkaDelta35 mutants were significantly smaller than those of rOka in melanoma cells. Infection of melanoma cells with rOkaDelta35 mutants was associated with disrupted cell fusion and polykaryocyte formation. The small plaque phenotype was not corrected by growth of rOkaDelta35 mutants in melanoma cells expressing the major VZV glycoprotein E, gE. The rOkaDelta35/35@Avr viruses displayed growth kinetics and plaque morphologies that were indistinguishable from those of rOka. Analysis with ORF35 Flag recombinants showed that the ORF35 gene product localized predominantly to the nuclei of infected cells. Evaluations in the SCIDhu mouse model demonstrated that ORF35 was required for efficient VZV infection of skin and T-cell xenografts, although the decrease in infectivity was most significant in skin. These mutagenesis experiments indicated that ORF35 was dispensable for VZV replication, but deleting ORF35 diminished growth in cultured cells and was associated with attenuated VZV infection of differentiated human skin and T cells in vivo. PMID- 15795268 TI - Role of low CD4 levels in the influence of human immunodeficiency virus type 1 envelope V1 and V2 regions on entry and spread in macrophages. AB - Human immunodeficiency virus type 1 (HIV-1) isolates vary in their ability to infect macrophages. Previous experiments have mapped viral determinants of macrophage infectivity to the V3 hypervariable region of the HIV-1 envelope glycoprotein. In our earlier studies, V1 and V2 sequences of HIV-1 were also shown to alter the ability of virus to spread in macrophage cultures, whereas no effect was seen in lymphocyte cultures. In the present study, determinants that allowed certain HIV-1 clones to infect and spread in macrophages were primarily mapped to the V2 region and were found to act by influencing early events of viral infection. By an assay of viral entry into macrophages, it was shown that viruses with the V2 region from the Ba-L strain of HIV-1 had >10-fold-higher entry efficiency than viruses with the V2 region derived from the NL4-3 strain. V1 region differences between these groups caused a twofold difference in entry. The known low expression of CD4 on macrophages appeared to be important in this process. In entry assays conducted with HeLa cell lines expressing various levels of CD4 and CCR5, low levels of CD4 influenced the efficiency of entry and fusion which were dependent on viral V1 and V2 envelope sequences. In contrast, no effect of V1 or V2 was seen in HeLa cells expressing high levels of CD4. Thus, the limited expression of CD4 on macrophages or other cell types could serve as a selective factor for V1 and V2 envelope sequences, and this selection could in turn influence many aspects of AIDS pathogenesis in vivo. PMID- 15795269 TI - Constitutive expression of alpha interferon by skin dendritic cells confers resistance to infection by foot-and-mouth disease virus. AB - The role of dendritic cells (DC) in the initiation of immune responses against foot-and-mouth disease virus (FMDV) is poorly understood. We analyzed the innate response of freshly isolated swine skin DC to the virus and show a rapid induction of beta interferon (IFN-beta) mRNA but not IFN-alpha mRNA. However, these DC secreted both IFN-alpha and IFN-beta proteins in response to live virus but not killed virus. Furthermore, the surface expression of swine major histocompatibility complex class II (SLA II) or CD80/CD86 molecules and antigen processing functions were not affected by FMDV exposure. Given the demonstrated sensitivity of FMDV to IFN-alpha/beta, there was no productive or nonproductive infection of these cells. Finally, freshly isolated skin DC constitutively expressed intracellular IFN-alpha protein in the absence of stimulation, with no detectable secretion of the cytokine until virus exposure. In situ analysis of these DC showed that these cells express and store IFN-alpha in uninfected animals. This is the first demonstration of the constitutive expression of IFN alpha in resident, tissue-derived DC and indicates that skin DC can play an important role in the innate immune response of swine to viral infections. PMID- 15795270 TI - The p92 polymerase coding region contains an internal RNA element required at an early step in Tombusvirus genome replication. AB - The replication of positive-strand RNA viral genomes involves various cis-acting RNA sequences. Generally, regulatory RNA sequences are present at or near genomic termini; however, internal replication elements (IREs) also exist. Here we report the structural and functional characterization of an IRE present in the readthrough portion of the p92 polymerase gene of Tomato bushy stunt virus. Analysis of this element in the context of a noncoding defective interfering RNA revealed a functional core structure composed of two noncontiguous segments of sequence that interact with each other to form an extended helical conformation. IRE activity required maintenance of several base-paired sections as well as two distinct structural features: (i) a short, highly conserved segment that can potentially form two different and mutually exclusive structures and (ii) an internal loop that contains a critical CC mismatch. The IRE was also shown to play an essential role within the context of the viral genome. In vivo analysis with novel RNA-based temperature-sensitive genomic mutants and translationally active subgenomic viral replicons revealed the following about the IRE: (i) it is active in the positive strand, (ii) it is dispensable late in the viral RNA replication process, and (iii) it is functionally inhibited by active translation over its sequence. Together, these results suggest that IRE activity is required in the cytosol at an early step in the viral replication process, such as template recruitment and/or replicase complex assembly. PMID- 15795271 TI - Specific binding of tombusvirus replication protein p33 to an internal replication element in the viral RNA is essential for replication. AB - The mechanism of template selection for genome replication in plus-strand RNA viruses is poorly understood. Using the prototypical tombusvirus, Tomato bushy stunt virus (TBSV), we show that recombinant p33 replicase protein binds specifically to an internal replication element (IRE) located within the p92 RNA dependent RNA polymerase coding region of the viral genome. Specific binding of p33 to the IRE in vitro depends on the presence of a C.C mismatch within a conserved RNA helix. Interestingly, the absence of the p33:p33/p92 interaction domain in p33 prevented specific but allowed nonspecific RNA binding, suggesting that a multimeric form of this protein is involved in the IRE-specific interaction. Further support for the selectivity of p33 binding in vitro was provided by the inability of the replicase proteins of the closely related Turnip crinkle virus and distantly related Hepatitis C virus to specifically recognize the TBSV IRE. Importantly, there was also a strong correlation between p33:IRE complex formation in vitro and viral replication in vivo, where mutations in the IRE that disrupted selective p33 binding in vitro also abolished TBSV RNA replication both in plant and in Saccharomyces cerevisiae cells. Based on these findings and the other known properties of p33 and the IRE, it is proposed that the p33:IRE interaction provides a mechanism to selectively recruit viral RNAs into cognate viral replicase complexes. Since all genera in Tombusviridae encode comparable replicase proteins, these results may be relevant to other members of this large virus family. PMID- 15795272 TI - Escape mutations alter proteasome processing of major histocompatibility complex class I-restricted epitopes in persistent hepatitis C virus infection. AB - Mutations in hepatitis C virus (HCV) genomes facilitate escape from virus specific CD8+ T lymphocytes in persistently infected chimpanzees. Our previous studies demonstrated that many of the amino acid substitutions in HCV epitopes prevented T-cell receptor recognition or binding to class I major histocompatibility complex molecules. Here we report that mutations within HCV epitopes also cause their destruction by changing the pattern of proteasome digestion. This mechanism of immune evasion provides further evidence of the potency of CD8+ T-cell selection pressure against HCV and should be considered when evaluating the significance of mutations in viral genomes from persistently infected chimpanzees and humans. PMID- 15795273 TI - Interleukin-15 increases effector memory CD8+ t cells and NK Cells in simian immunodeficiency virus-infected macaques. AB - Interleukin-15 (IL-15) in vitro treatment of peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus (HIV)-infected individuals specifically enhances the function and survival of HIV-specific CD8+ T cells, while in vivo IL 15 treatment of mice preferentially expands memory CD8+ T cells. In this study, we investigated the in vivo effect of IL-15 treatment in 9 SIVmac251-infected cynomolgus macaques (low dose of IL-15, 10 microg/kg of body weight, n = 3; high dose of IL-15, 100 microg/kg, n = 3; control [saline], n = 3; dose administered twice weekly for 4 weeks). IL-15 treatment induced a nearly threefold increase in peripheral blood CD8+CD3- NK cells. Furthermore, CD8+ T-cell numbers increased more than twofold, mainly due to an increase in the CD45RA-CD62L- and CD45RA+CD62L- effector memory CD8+ T cells. Expression of Ki-67 in the CD8+ T cells indicated expansion of CD8+ T cells and not redistribution. IL-15 did not affect CD4+ T-cell, B-cell, and CD14+ macrophage numbers. No statistically significant differences in changes from baseline in the viral load were observed when control-, low-dose-, and high-dose-treated animals were compared. No clinical adverse effects were observed in any of the animals studied. The selective expansion of effector memory CD8+ T cells and NK cells by IL-15 further supports IL-15's possible therapeutic use in viral infections such as HIV infection. PMID- 15795274 TI - Retroviral recombination in vivo: viral replication patterns and genetic structure of simian immunodeficiency virus (SIV) populations in rhesus macaques after simultaneous or sequential intravaginal inoculation with SIVmac239Deltavpx/Deltavpr and SIVmac239Deltanef. AB - To characterize the occurrence, frequency, and kinetics of retroviral recombination in vivo, we intravaginally inoculated rhesus macaques, either simultaneously or sequentially, with attenuated simian immunodeficiency virus (SIV) strains having complementary deletions in their accessory genes and various degrees of replication impairment. In monkeys inoculated simultaneously with SIVmac239Deltavpx/Deltavpr and SIVmac239Deltanef, recombinant wild-type (wt) virus and wild-type levels of plasma viral RNA (vRNA) were detected in blood by 2 weeks postinoculation. In monkeys inoculated first with SIVmac239Deltavpx/Deltavpr and then with SIVmac239Deltanef, recombination occurred but was associated with lower plasma vRNA levels than plasma vRNA levels seen for monkeys inoculated intravaginally with wt SIVmac239. In one monkey, recombination occurred 6 weeks after the challenge with SIVmac239Deltanef when plasma SIVmac239Deltavpx/Deltavpr RNA levels were undetectable. In monkeys inoculated first with the more highly replicating strain, SIVmac239Deltanef, and then with SIVmac239Deltavpx/Deltavpr, wild-type recombinant virus was not detected in blood or tissues. Instead, a virus that had repaired the deletion in the nef gene by a compensatory mutation was found in one animal. Overall, recombinant SIV was eventually found in four of six animals intravaginally inoculated with the two SIVmac239 deletion mutants. These findings show that recombination can occur readily in vivo after mucosal SIV exposure and thus contributes to the generation of viral genetic diversity and enhancement of viral fitness. PMID- 15795275 TI - CD4+ T-cell responses to Epstein-Barr virus (EBV) latent-cycle antigens and the recognition of EBV-transformed lymphoblastoid cell lines. AB - There is considerable interest in the potential of Epstein-Barr virus (EBV) latent antigen-specific CD4+ T cells to act as direct effectors controlling EBV induced B lymphoproliferations. Such activity would require direct CD4+ T-cell recognition of latently infected cells through epitopes derived from endogenously expressed viral proteins and presented on the target cell surface in association with HLA class II molecules. It is therefore important to know how often these conditions are met. Here we provide CD4+ epitope maps for four EBV nuclear antigens, EBNA1, -2, -3A, and -3C, and establish CD4+ T-cell clones against 12 representative epitopes. For each epitope we identify the relevant HLA class II restricting allele and determine the efficiency with which epitope-specific effectors recognize the autologous EBV-transformed B-lymphoblastoid cell line (LCL). The level of recognition measured by gamma interferon release was consistent among clones to the same epitope but varied between epitopes, with values ranging from 0 to 35% of the maximum seen against the epitope peptide loaded LCL. These epitope-specific differences, also apparent in short-term cytotoxicity and longer-term outgrowth assays on LCL targets, did not relate to the identity of the source antigen and could not be explained by the different functional avidities of the CD4+ clones; rather, they appeared to reflect different levels of epitope display at the LCL surface. Thus, while CD4+ T-cell responses are detectable against many epitopes in EBV latent proteins, only a minority of these responses are likely to have therapeutic potential as effectors directly recognizing latently infected target cells. PMID- 15795276 TI - Human immunodeficiency virus (HIV)-specific gamma interferon secretion directed against all expressed HIV genes: relationship to rate of CD4 decline. AB - Immune responses to human immunodeficiency virus (HIV) are detected at all stages of infection and are believed to be responsible for controlling viremia. This study seeks to determine whether gamma interferon (IFN-gamma)-secreting HIV specific T-cell responses influence disease progression as defined by the rate of CD4 decline. The study population consisted of 31 subjects naive to antiretroviral therapy. All were monitored clinically for a median of 24 months after the time they were tested for HIV-specific responses. The rate of CD4+-T cell loss was calculated for all participants from monthly CD4 counts. Within this population, 17 subjects were classified as typical progressors, 6 subjects were classified as fast progressors, and 8 subjects were classified as slow progressors. Peripheral blood mononuclear cells were screened for HIV-specific IFN-gamma responses to all expressed HIV genes. Among the detected immune responses, 48% of the recognized peptides were encoded by Gag and 19% were encoded by Nef gene products. Neither the breadth nor the magnitude of HIV specific responses correlated with the viral load or rate of CD4 decline. The breadth and magnitude of HIV-specific responses did not differ significantly among typical, fast, and slow progressors. These results support the conclusion that although diverse HIV-specific IFN-gamma-secreting responses are mounted during the asymptomatic phase, these responses do not seem to modulate disease progression rates. PMID- 15795277 TI - Induction of the human papillomavirus type 31 late promoter requires differentiation but not DNA amplification. AB - The human papillomavirus (HPV) life cycle is linked to the differentiation state of the host cell. In virus-infected undifferentiated basal epithelial cells, HPV genomes are maintained as episomes at low copy number. Upon differentiation, a concomitant increase in viral copy number and an induction of late gene expression from a differentiation-specific promoter is seen. To investigate whether late gene expression was dependent on the amplification of the viral genome, inhibitors of DNA replication and in vitro systems for epithelial differentiation were used in conjunction with cells that stably maintain HPV31 episomes. Treatment of cells induced to differentiate in methylcellulose with the DNA synthesis inhibitor cytosine beta-arabinofuranoside (AraC) blocked viral DNA amplification but did not prevent induction of late transcription. This suggests that late gene expression does not strictly require amplification of the viral genome and that differentiation signals alone are sufficient to activate transcription from the late promoter. However, DNA amplification does appear to be necessary for maximal induction of the late promoter. In order to examine the cis-acting elements that contribute to the activation of the late promoter, a transient reporter assay was developed. In these assays, an induction of late gene expression was seen upon differentiation that was specific to the late promoter. Mapping studies localized important regulatory elements to the E6/E7 region and identified short sequences that could serve as binding sites for transcription factors. Elements within the upstream regulatory region were also found to positively and negatively influence transcription from the late promoter. These results identify mechanisms important for the differentiation dependent activation of late gene expression of high-risk papillomaviruses. PMID- 15795278 TI - Induction of humoral immune responses following vaccination with envelope containing, formaldehyde-treated, thermally inactivated human immunodeficiency virus type 1. AB - The lack of success of subunit human immunodeficiency virus type 1 (HIV-1) vaccines to date suggests that multiple components or a complex virion structure may be required. We previously demonstrated retention of the major conformational epitopes of HIV-1 envelope following thermal treatment of virions. Moreover, antibody binding to some of these epitopes was significantly enhanced following thermal treatment. These included the neutralizing epitopes identified by monoclonal antibodies 1b12, 2G12, and 17b, some of which have been postulated to be partially occluded or cryptic in native virions. Based upon this finding, we hypothesized that a killed HIV vaccine could be derived to elicit protective humoral immune responses. Shedding of HIV-1 envelope has been described for some strains of HIV-1 and has been cited as one of the major impediments to developing an inactivated HIV-1 vaccine. In the present study, we demonstrate that treatment of virions with low-dose formaldehyde prior to thermal inactivation retains the association of viral envelope with virions. Moreover, mice and nonhuman primates vaccinated with formaldehyde-treated, thermally inactivated virions produce antibodies capable of neutralizing heterologous strains of HIV in peripheral blood mononuclear cell-, MAGI cell-, and U87-based infectivity assays. These data indicate that it is possible to create an immunogen by using formaldehyde treated, thermally inactivated HIV-1 virions to induce neutralizing antibodies. These findings have broad implications for vaccine development. PMID- 15795279 TI - Latent membrane protein 1 regulates STAT1 through NF-kappaB-dependent interferon secretion in Epstein-Barr virus-immortalized B cells. AB - Constitutive activation of signal transducer and activator of transcription 1 (STAT1) is a distinctive feature of Epstein-Barr virus (EBV)-immortalized B cells (lymphoblastoid cell lines [LCLs]). The expression of STAT1 in these cells is modulated by the latent membrane protein 1 (LMP1), but the mechanism of STAT1 activation has remained unclear. We demonstrate that the tyrosine phosphorylation of STAT1 in LCLs results from an indirect pathway encompassing an NF-kappaB dependent secretion of interferons (IFNs). The cell culture supernatant of LCLs induced tyrosine phosphorylation of STAT1 in cells with no constitutively activated STAT1. Moreover, removal of supernatant from LCLs was sufficient to decrease the phosphorylation of STAT1. Inhibition of NF-kappaB activity by different pharmacological inhibitors (i.e., parthenolide, MG132 and BAY 11-7082) and by overexpressed mutated IkappaBalpha prevented the activation of STAT1. To identify the factors involved, we performed macroarray cDNA profiling with or without inhibition of NF-kappaB. The expression of several cytokines was NF kappaB dependent among those alpha and gamma IFNs (IFN-alpha and IFN-gamma), known activators of STAT1. By real-time PCR and enzyme-linked immunosorbent assay we show that IFN-alpha and IFN-gamma are expressed and released by LCLs in an NF kappaB-dependent manner. Finally, the blocking of the IFN-alpha and IFN-gamma by neutralizing antibodies led to the complete inhibition of tyrosine phosphorylation of STAT1. Taken together, our results clearly show that LMP1 induced tyrosine phosphorylation of STAT1 is almost exclusively due to the NF kappaB-dependent secretion of IFNs. Whether this response, which is usually considered to be antiviral, is in fact required for the persistence of the virus remains to be elucidated. PMID- 15795280 TI - CD4-independent entry and replication of simian immunodeficiency virus in primary rhesus macaque astrocytes are regulated by the transmembrane protein. AB - Previous studies have demonstrated that the genetic determinants of simian immunodeficiency virus (SIV) neurovirulence map to the env and nef genes. Recent studies from our laboratory demonstrated that SIV replication in primary rhesus macaque astrocyte cultures is dependent upon the nef gene. Here, we demonstrate that macrophage tropism is not sufficient for replication in astrocytes and that specific amino acids in the transmembrane (TM) portion of Env are also important for optimal SIV replication in astrocytes. Specifically, a Gly at amino acid position 751 and truncation of the cytoplasmic tail of TM are required for efficient replication in these cells. Studies using soluble CD4 demonstrated that these changes within the TM protein regulate CD4-independent, CCR5-dependent entry of virus into astrocytes. In addition, we observed that two distinct CD4 independent, neuroinvasive strains of SIV/DeltaB670 also replicated efficiently in astrocytes, further supporting the role of CD4 independence as an important determinant of SIV infection of astrocytes in vitro and in vivo. PMID- 15795281 TI - Host and viral proteins in the virion of Kaposi's sarcoma-associated herpesvirus. AB - Infection of cultured cells with Kaposi's sarcoma associated herpesvirus (KSHV) typically establishes a latent infection, in which only a few viral genes are expressed. Recently, it has been reported that a subset of lytic genes are transiently expressed very early after viral entry but that this burst of abortive lytic gene expression is terminated with the supervention of latency (H. H. Krishnan, P. P. Naranatt, M. S. Smith, L. Zeng, C. Bloomer, and B. Chandran, J. Virol. 78:3601-3620, 2004). To identify molecules imported into cells by KSHV that might influence this gene expression program, we have examined the protein composition of the KSHV particle. Immunoblotting of virus particles demonstrated that RTA, the lytic switch protein, and RAP, a viral protein that is a transcriptional and cell cycle modulator, were both incorporated into virus particles. In a second approach, polypeptides isolated from purified virions were identified by mass-spectrometric analysis of their constituent tryptic peptides. With this approach we were able to identify 18 major virion proteins, including structural, regulatory, and signaling proteins of both viral and cellular origin. PMID- 15795282 TI - Preferential feline immunodeficiency virus (FIV) infection of CD4+ CD25+ T regulatory cells correlates both with surface expression of CXCR4 and activation of FIV long terminal repeat binding cellular transcriptional factors. AB - Previously, we have characterized feline CD4+ CD25+ T-regulatory (Treg) cells with regard to their immune regulatory properties and ability to support feline immunodeficiency virus (FIV) replication in vitro and in vivo. Our studies showed that while CD4+ CD25+ cells were capable of replicating FIV in the presence of interleukin-2 (IL-2) alone, CD4+ CD25- cells harbored a latent infection that required a strong mitogenic stimulus to activate virus replication. In the present study, we investigated the mechanisms governing the preferential replication of FIV in highly purified CD4+ CD25+ Treg cells compared to their CD4+ CD25+ counterparts. Studies aimed at elucidating mechanisms regulating infection of these cells revealed that CD4+ CD25- cells were less susceptible to FIV binding and entry than CD4+ CD25+ cells, which correlated with increased surface expression of FIV coreceptor CXCR4. In addition, the number of CD4+ CD25+ cells that expressed the primary receptor CD134 was greater than for CD4+ CD25- cells. Although increased permissiveness to FIV infection of CD4+ CD25- cells following mitogenic stimulation correlated strongly with upregulation of surface CXCR4, it did not correlate with CD134 expression. Further, study of intracellular factors regulating FIV replication revealed that CD4+ CD25+ but not CD4+ CD25- T cells showed constitutive and IL-2-responsive transactivation of activating transcription factor, CAAT enhancer binding protein, and activating protein 1 transcription factors that are important for FIV replication. These factors were upregulated in CD4+ CD25- T cells following ConA stimulation, which correlated with FIV replication. This is the first report elucidating the mechanisms that allow for productive lentiviral infection of CD4+ CD25+ Treg cells. PMID- 15795283 TI - Evidence of recombination in the norovirus capsid gene. AB - Noroviruses are single-stranded RNA viruses with high genomic variability. They have emerged in the last decade as a major cause of acute gastroenteritis. It remains so far unclear whether norovirus evolution is driven by sequence mutation and/or recombination. In this study, we have assessed the occurrence of recombination in the norovirus capsid gene. For this purpose, 69 complete capsid sequences of norovirus strains accessible in GenBank as well as 25 complete capsid sequences generated from norovirus-positive clinical samples were examined. Unreported recombination was detected in about 8% of norovirus strains belonging to genetic clusters I/1 (n = 1), II/1 (n = 1), II/3 (n = 1), II/4 (n = 3), and II/5 (n = 1). Recombination breakpoints were mainly located at the interface of the putative P1-1 and P2 domains of the capsid protein and/or within the P2 domain. The recombination region displayed features such as length, sequence composition (upstream and downstream GC- and AU-rich sequences, respectively), and predicted RNA secondary structure that are characteristic of homologous recombination activators. Our results suggest that recombination in the norovirus capsid gene may naturally occur, involving capsid domains presumably exposed to immunological pressure. PMID- 15795284 TI - Enfuvirtide resistance mutations: impact on human immunodeficiency virus envelope function, entry inhibitor sensitivity, and virus neutralization. AB - Enfuvirtide (ENF/T-20/Fuzeon), the first human immunodeficiency virus (HIV) entry inhibitor to be licensed, targets a structural intermediate of the entry process. ENF binds the HR1 domain in gp41 after Env has bound CD4, preventing conformational changes needed for membrane fusion. Mutations in HR1 that confer ENF resistance can arise following ENF therapy. ENF resistance mutations were introduced into an R5- and X4-tropic Env to examine their impact on fusion, infection, and sensitivity to different classes of entry inhibitors and neutralizing antibodies. HR1 mutations could reduce infection and fusion efficiency and also delay fusion kinetics, likely accounting for their negative impact on viral fitness. HR1 mutations had minimal effect on virus sensitivity to other classes of entry inhibitors, including those targeting CD4 binding (BMS-806 and a CD4-specific monoclonal antibody [MAb]), coreceptor binding (CXCR4 inhibitor AMD3100 and CCR5 inhibitor TAK-779), or fusion (T-1249), indicating that ENF-resistant viruses can remain sensitive to other entry inhibitors in vivo. Some HR1 mutations conferred increased sensitivity to a subset of neutralizing MAbs that likely target fusion intermediates or with epitopes preferentially exposed following receptor interactions (17b, 48D, 2F5, 4E10, and IgGb12), as well as sera from some HIV-positive individuals. Mechanistically, enhanced neutralization correlated with reduced fusion kinetics, indicating that, in addition to steric constraints, kinetics may also limit virus neutralization by some antibodies. Therefore, escape from ENF comes at a cost to viral fitness and may confer enhanced sensitivity to humoral immunity due to prolonged exposure of epitopes that are not readily accessible in the native Env trimer. Resistance to other entry inhibitors was not observed. PMID- 15795285 TI - The majority of currently circulating human immunodeficiency virus type 1 clade B viruses fail to prime cytotoxic T-lymphocyte responses against an otherwise immunodominant HLA-A2-restricted epitope: implications for vaccine design. AB - Human immunodeficiency virus type 1 (HIV-1) mutates to escape immune selection pressure, but there is little evidence of selection mediated through HLA-A2, the dominant class I allele in persons infected with clade B virus. Moreover, HLA-A2 restricted responses are largely absent in the acute phase of infection as the viral load is being reduced, suggesting that circulating viruses may lack immunodominant epitopes targeted through HLA-A2. Here we demonstrate an A2 restricted epitope within Vpr (Vpr59-67) that is targeted by acute-phase HIV-1 specific CD8+ T cells, but only in a subset of persons expressing HLA-A2. Individuals in the acute stage of infection with viruses containing the most common current sequence within this epitope (consensus sequence) were unable to mount epitope-specific T-cell responses, whereas subjects infected with the less frequent I60L variant all developed these responses. The I60L variant epitope was a stronger binder to HLA-A2 and was recognized by epitope-specific T cells at lower peptide concentrations than the consensus sequence epitope. These data demonstrate that HLA-A2 is capable of contributing to the acute-phase cytotoxic T lymphocyte response in infected subjects, but that most currently circulating viruses lack a dominant immunogenic epitope presented by this allele, and suggest that immunodominant epitopes restricted by common HLA alleles may be lost as the epidemic matures. PMID- 15795287 TI - Penetration of enveloped double-stranded RNA bacteriophages phi13 and phi6 into Pseudomonas syringae cells. AB - Bacteriophages phi6 and phi13 are related enveloped double-stranded RNA viruses that infect gram-negative Pseudomonas syringae cells. phi6 uses a pilus as a receptor, and phi13 attaches to the host lipopolysaccharide. We compared the entry-related events of these two viruses, including receptor binding, envelope fusion, peptidoglycan penetration, and passage through the plasma membrane. The infection-related events are dependent on the multiplicity of infection in the case of phi13 but not with phi6. A temporal increase of host outer membrane permeability to lipophilic ions was observed from 1.5 to 4 min postinfection in both virus infections. This enhanced permeability period coincided with the fast dilution of octadecyl rhodamine B-labeled virus-associated lipid molecules. This result is in agreement with membrane fusion, and the presence of temporal virus derived membrane patches on the outer membrane. Similar to phi6, phi13 contains a thermosensitive lytic enzyme involved in peptidoglycan penetration. The phage entry also caused a limited depolarization of the plasma membrane. Inhibition of host respiration considerably decreased the efficiency of irreversible virus binding and membrane fusion. An active role of cell energy metabolism in restoring the infection-induced defects in the cell envelope was also observed. PMID- 15795286 TI - Hepatitis C virus NS5A-mediated activation of phosphoinositide 3-kinase results in stabilization of cellular beta-catenin and stimulation of beta-catenin responsive transcription. AB - The hepatitis C virus (HCV) nonstructural NS5A protein has been shown to bind to and activate phosphoinositide 3-kinase (PI3K), resulting in activation of the downstream effector serine/threonine kinase Akt/protein kinase B. Here we present data pertaining to the effects of NS5A-mediated Akt activation on its downstream targets. Using a recombinant baculovirus to deliver the complete HCV polyprotein to human hepatoma cells in a tetracycline-regulable fashion, we confirm that expression of the complete HCV polyprotein also activates PI3K and Akt. We further show that this results in the inhibition of the Akt substrate Forkhead transcription factor and the stimulation of phosphorylation of a second key Akt substrate, glycogen synthase kinase-3beta (GSK-3beta). Phosphorylation of GSK 3beta results in its inactivation; consistent with this, we show that expression of the HCV polyprotein results in the accumulation of beta-catenin. Finally, we show that levels of beta-catenin-dependent transcription are also elevated in the presence of the HCV polyprotein. Given the prevalence of beta-catenin mutations in many human tumors, especially colon and hepatocellular carcinomas, these data implicate NS5A-mediated PI3K activation as a contributory factor in the increasingly common association between HCV infection and the development of hepatocellular carcinoma. PMID- 15795288 TI - Characterization of the elements and proteins responsible for interferon stimulated gene induction by human cytomegalovirus. AB - Human cytomegalovirus (HCMV) infection of human fibroblast cells activates a large number of interferon-stimulated genes (ISGs) in a viral envelope-cell membrane fusion-dependent mechanism. In this study, we identified two interferon response elements, the interferon-stimulated response element (ISRE) and the gamma interferon-activated site (GAS), which act as HCMV response sites (VRS). Gel mobility shift assays showed that cellular proteins form specific and identical complexes with ISRE and GAS elements, and the binding of these complexes to ISRE and GAS is stimulated by HCMV infection. Point mutations in the consensus sequences of ISRE and GAS completely abolished their activities in response to HCMV-mediated transactivation, as well as their abilities to interact with HCMV-activated VRS-binding proteins. Interferon regulatory factor 3 does not appear to be present in the VRS-binding complexes or to be involved directly in HCMV-mediated ISG activation. Using ProteinChip technology, four potential proteins were identified, ranging from 20 to 42 kDa, in the VRS-binding complexes. The data suggest that HCMV infection activates VRS-binding proteins, which then bind to the VRS and stimulate ISG expression. PMID- 15795289 TI - Two gamma interferon-activated site-like elements in the human cytomegalovirus major immediate-early promoter/enhancer are important for viral replication. AB - Human cytomegalovirus (HCMV) infection directly initiates a signal transduction pathway that leads to activation of a large number of cellular interferon stimulated genes (ISGs). Our previous studies demonstrated that two interferon response elements, the interferon-stimulated response element and gamma interferon-activated site (GAS), in the ISG promoters serve as HCMV response sites (VRS). Interestingly, two GAS-like VRS elements (VRS1) were also present in the HCMV major immediate-early promoter-enhancer (MIEP/E). In this study, the importance of these VRS elements in viral replication was investigated. We demonstrate that the expression of the major IE genes, IE1 and IE2, is interferon inducible. To understand the biological significance of this signal transduction pathway in HCMV major IE expression, the two VRS1 in the MIEP/E were mutated. Mutant HCMVs in which the VRS elements were deleted or that contained point mutations grew dramatically more slowly than wild-type virus at a low multiplicity of infection (MOI). Insertion of wild-type VRS1 into the mutant viral genome rescued the slow growth phenotype. Furthermore, the expression levels of major IE RNAs and proteins were greatly reduced during infection with the VRS mutants at a low MOI. HCMV microarray analysis indicated that infection of host cells with the VRS mutant virus resulted in a global reduction in the expression of viral genes. Collectively, these data demonstrate that the two VRS elements in the MIEP/E are necessary for efficient viral gene expression and replication. This study suggests that although the HCMV-initiated signal transduction pathway results in induction of cellular antiviral genes, it also functions to stimulate viral major IE gene expression. This might be a new viral strategy in which the pathway is used to regulate gene expression and play a role in reactivation. PMID- 15795291 TI - The murine gammaherpesvirus 68 ORF27 gene product contributes to intercellular viral spread. AB - Herpesviruses remain predominantly cell associated within their hosts, implying that they spread between cells by a mechanism distinct from free virion release. We previously identified the efficient release of murine gammaherpesvirus 68 (MHV 68) virions as a function of the viral gp150 protein. Here we show that the MHV 68 ORF27 gene product, gp48, contributes to the direct spread of viruses from lytically infected to uninfected cells. Monoclonal antibodies to gp48 identified it on infected cell surfaces and in virions. gp48-deficient viruses showed no obvious deficit in virion cell binding, single-cycle replication, or virion release but had reduced lytic propagation between cells. After intranasal infection of mice, ORF27-deficient viruses were impaired predominantly in lytic replication in the lungs. There was a small deficit in latency establishment, but long-term latency appeared normal. Since ORF27 has homologs in both Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus, it is likely part of a conserved mechanism employed by gammaherpesviruses to disseminate lytically in their hosts. PMID- 15795290 TI - Structure of adeno-associated virus type 4. AB - Adeno-associated virus (AAV) is a member of the Parvoviridae, belonging to the Dependovirus genus. Currently, several distinct isolates of AAV are in development for use in human gene therapy applications due to their ability to transduce different target cells. The need to manipulate AAV capsids for specific tissue delivery has generated interest in understanding their capsid structures. The structure of AAV type 4 (AAV4), one of the most antigenically distinct serotypes, was determined to 13-A resolution by cryo-electron microscopy and image reconstruction. A pseudoatomic model was built for the AAV4 capsid by use of a structure-based sequence alignment of its major capsid protein, VP3, with that of AAV2, to which AAV4 is 58% identical and constrained by its reconstructed density envelope. The model showed variations in the surface loops that may account for the differences in receptor binding and antigenicity between AAV2 and AAV4. The AAV4 capsid surface topology also shows an unpredicted structural similarity to that of Aleutian mink disease virus and human parvovirus B19, autonomous members of the genus, despite limited sequence homology. PMID- 15795292 TI - Regions of the varicella-zoster virus open reading frame 63 latency-associated protein important for replication in vitro are also critical for efficient establishment of latency. AB - Varicella-zoster virus (VZV) open reading frame 63 (ORF63) is one of the most abundant transcripts expressed during VZV latency in humans, and ORF63 protein has been detected in human ganglia by several laboratories. Deletion of over 90% of the ORF63 gene showed that the protein is required for efficient establishment of latency in rodents. We have constructed viruses with a series of mutations in ORF63. While prior experiments showed that transfection of cells with a plasmid expressing ORF63 but lacking the putative nuclear localization signal of the protein resulted in increased expression of the protein in the cytoplasm, we found that ORF63 protein remained in the nucleus in cells infected with a VZV ORF63 nuclear localization signal deletion mutant. This mutant was not impaired for growth in cell culture or for latency in rodents. Replacement of five serine or threonine phosphorylation sites in ORF63 with alanines resulted in a virus that was impaired for replication in vitro and for latency. A series of ORF63 carboxy-terminal mutants showed that the last 70 amino acids do not affect replication in vitro or latency in rodents; however, the last 108 amino acids are important for replication and latency. Thus, regions of ORF63 that are important for replication in vitro are also required for efficient establishment of latency. PMID- 15795293 TI - ND10 components relocate to sites associated with herpes simplex virus type 1 nucleoprotein complexes during virus infection. AB - Infections with DNA viruses commonly result in the association of viral genomes and replication compartments with cellular nuclear substructures known as promyelocytic leukemia protein (PML) nuclear bodies or ND10. While there is evidence that viral genomes can associate with preexisting ND10, we demonstrate in this study by live-cell microscopy that structures resembling ND10 form de novo and in association with viral genome complexes during the initial stages of herpes simplex virus type 1 (HSV-1) infection. Consistent with previous studies, we found that the major ND10 proteins PML, Sp100, and hDaxx are exchanged very rapidly between ND10 foci and the surrounding nucleoplasm in live cells. The dynamic nature of the individual protein molecule components of ND10 provides a mechanism by which ND10 proteins can be recruited to novel sites during virus infection. These observations explain why the genomes and replication compartments of DNA viruses that replicate in the cell nucleus are so commonly found in association with ND10. These findings are discussed with reference to the nature, location, and potential number of HSV-1 prereplication compartments and to the dynamic aspects of HSV-1 genomes and viral products during the early stages of lytic infection. PMID- 15795294 TI - Development and assessment of human adenovirus type 11 as a gene transfer vector. AB - Adenovirus vectors based on human serotype 5 (Ad5) have successfully been used as gene transfer vectors in many gene therapy-based approaches to treat disease. Despite their widespread application, many potential therapeutic applications are limited by the widespread prevalence of vector-neutralizing antibodies within the human population and the inability of Ad5-based vectors to transduce important therapeutic target cell types. In an attempt to circumvent these problems, we have developed Ad vectors based on human Ad serotype 11 (Ad11), since the prevalence of neutralizing antibodies to Ad11 in humans is low. E1-deleted Ad11 vector genomes were generated by homologous recombination in 293 cells expressing the Ad11-E1B55K protein or by recombination in Escherichia coli. E1-deleted Ad11 genomes did not display transforming activity in rodent cells. Transduction of primary human CD34+ hematopoietic progenitor cells and immature dendritic cells was more efficient with Ad11 vectors than with Ad5 vectors. Thirty minutes after intravenous injection into mice that express one of the Ad11 receptors (CD46), we found, in a pattern and at a level comparable to what is found in humans, Ad11 vector genomes in all analyzed organs, with the highest amounts in liver, lung, kidney, and spleen. Neither Ad11 genomes nor Ad11 vector-mediated transgene expression were, however, detected at 72 h postinfusion. A large number of Ad11 particles were also found to be associated with circulating blood cells. We also discovered differences in in vitro transduction efficiencies and in vivo biodistributions between Ad11 vectors and chimeric Ad5 vectors possessing Ad11 fibers, indicating that Ad11 capsid proteins other than fibers influence viral infectivity and tropism. Overall, our study provides a basis for the application of Ad11 vectors for in vitro and in vivo gene transfer and for gaining an understanding of the factors that determine Ad tropism. PMID- 15795295 TI - Expression of the full-length form of gp2 of equine herpesvirus 1 (EHV-1) completely restores respiratory virulence to the attenuated EHV-1 strain KyA in CBA mice. AB - Wild-type equine herpesvirus 1 (EHV-1) strains express a large (250-kDa) glycoprotein, gp2, that is encoded by EUs4 (gene 71) located within the unique short region of the genome. DNA sequence analysis revealed that EUs4 of the pathogenic EHV-1 strain RacL11 is an open reading frame of 2,376 bp that encodes a protein of 791 amino acids. The attenuated EHV-1 vaccine strain KyA harbors an in-frame deletion of 1,242 bp from bp 222 to 1461 and expresses a truncated gp2 of 383 amino acids. To determine the relative contribution of gp2 to EHV-1 pathogenesis, we compared the course of respiratory infection of CBA mice infected with either wild-type RacL11, attenuated KyA, or a recombinant KyA that expresses the full-length gp2 protein (KyARgp2F). Mice infected with KyA lost a negligible amount of body weight (0.18% total weight loss) on day 1 postinfection and regained weight thereafter, whereas mice infected with KyARgp2F or RacL11 steadily lost weight beginning on day 1 and experienced a 20 and 18% loss in body weight, respectively, by day 3. Immunohistochemical and flow cytometric analyses revealed higher numbers of T and B lymphocytes and an extensive consolidation consisting of large numbers of Mac-1-positive cells in the lungs of animals infected with KyARgp2F compared to animals infected with KyA. RNase protection analyses revealed increased expression of numerous cytokines and chemokines, including interleukin-1beta (IL-1beta), IL-6, tumor necrosis factor alpha, macrophage inflammatory protein 1alpha (MIP-1alpha), MIP-1beta, MIP-2, interferon gamma-inducible protein, monocyte chemotactic protein 1, and T-cell activation gene 3 at 12 h postinfection with KyARgp2F. Three independent DNA array experiments confirmed these results and showed a 2- to 13-fold increase in the expression of 31 inflammatory genes at 8 and 12 h postinfection with KyARgp2F compared to infection with KyA. Taken together, the results indicate that expression of full-length gp2 is sufficient to restore full respiratory virulence to the attenuated KyA strain and raise caution concerning the inclusion of full length gp2 in the development of EHV-1 vaccines. PMID- 15795296 TI - Inhibition of dengue virus serotypes 1 to 4 in vero cell cultures with morpholino oligomers. AB - Five dengue (DEN) virus-specific R5F2R4 peptide-conjugated phosphorodiamidate morpholino oligomers (P4-PMOs) were evaluated for their ability to inhibit replication of DEN virus serotype 2 (DEN-2 virus) in mammalian cell culture. Initial growth curves of DEN-2 virus 16681 were obtained in Vero cells incubated with 20 microM P4-PMO compounds. At 6 days after infection, a P4-PMO targeting the 3'-terminal nucleotides of the DEN-2 virus genome and a random-sequence P4 PMO showed relatively little suppression of DEN-2 virus titer (0.1 and 0.9 log10, respectively). P4-PMOs targeting the AUG translation start site region of the single open reading frame and the 5' cyclization sequence region had moderate activity, generating 1.6- and 1.8-log10 reductions. Two P4-PMO compounds, 5'SL and 3'CS (targeting the 5'-terminal nucleotides and the 3' cyclization sequence region, respectively), were highly efficacious, each reducing the viral titer by greater than 5.7 log10 compared to controls at 6 days after infection with DEN-2 virus. Further experiments showed that 5'SL and 3'CS inhibited DEN-2 virus replication in a dose-dependent and sequence-specific manner. Treatment with 10 microM 3'CS reduced the titers of all four DEN virus serotypes, i.e., DEN-1 (strain 16007), DEN-2 (16681), DEN-3 (16562), and DEN-4 (1036) viruses by over 4 log10, in most cases to below detectable limits. The extent of 3'CS efficacy was affected by the timing of compound application in relation to viral infection of the cells. The 5'SL and 3'CS P4-PMOs did not suppress the replication of West Nile virus NY99 in Vero cells. These data indicate that further evaluation of the 5'SL and 3'CS compounds as potential DEN virus therapeutics is warranted. PMID- 15795298 TI - A peptide pertaining to the loop segment of human immunodeficiency virus gp41 binds and interacts with model biomembranes: implications for the fusion mechanism. AB - The human immunodeficiency virus gp41 envelope protein mediates the entry of the virus into the target cell by promoting membrane fusion. In order to gain new insights into the viral fusion mechanism, we studied a 35-residue peptide pertaining to the loop domain of gp41, both in solution and membrane bound, by using infrared and fluorescence spectroscopy. We show here that the peptide, which has a membrane-interacting surface, binds and interacts with phospholipid model membranes and tends to aggregate in the presence of a membranous medium and induce the leakage of vesicle contents. The results reported in this work, i.e., the destabilization and fusion of negatively charged model membranes, suggest an essential role of the loop domain in the membrane fusion process induced by gp41. PMID- 15795297 TI - Murine gammaherpesvirus 68 open reading frame 45 plays an essential role during the immediate-early phase of viral replication. AB - Murine gammaherpesvirus 68 (MHV-68) has been developed as a model for the human gammaherpesviruses Epstein-Barr virus and human herpesvirus 8/Kaposi's sarcoma associated herpesvirus (HHV-8/KSHV), which are associated with several types of human diseases. Open reading frame 45 (ORF45) is conserved among the members of the Gammaherpesvirinae subfamily and has been suggested to be a virion tegument protein. The repression of ORF45 expression by small interfering RNAs inhibits MHV-68 viral replication. However, the gene product of MHV-68 ORF45 and its function have not yet been well characterized. In this report, we show that MHV 68 ORF45 is a phosphorylated nuclear protein. We constructed an ORF45-null MHV-68 mutant virus (45STOP) by the insertion of translation termination codons into the portion of the gene encoding the N terminus of ORF45. We demonstrated that the ORF45 protein is essential for viral gene expression immediately after the viral genome enters the nucleus. These defects in viral replication were rescued by providing ORF45 in trans or in an ORF45-null revertant (45STOP.R) virus. Using a transcomplementation assay, we showed that the function of ORF45 in viral replication is conserved with that of its KSHV homologue. Finally, we found that the C-terminal 23 amino acids that are highly conserved among the Gammaherpesvirinae subfamily are critical for the function of ORF45 in viral replication. PMID- 15795299 TI - Plateau levels of viremia correlate with the degree of CD4+-T-cell loss in simian immunodeficiency virus SIVagm-infected pigtailed macaques: variable pathogenicity of natural SIVagm isolates. AB - Simian immunodeficiency virus from African green monkeys (SIVagm) results in asymptomatic infection in its natural host species. The virus is not inherently apathogenic, since infection of pigtailed (PT) macaques (Macaca nemestrina) with one isolate of SIVagm results in an immunodeficiency syndrome characterized by progressive CD4+-T-cell depletion and opportunistic infections. This virus was passaged once in a PT macaque and, thus, may not be entirely reflective of the virulence of the parental strain. The goal of the present study was to assess the pathogenicity of the PT-passaged isolate (SIVagm9063) and two primary SIVagm isolates in PT macaques, including the parental strain of the PT-passaged variant. Infection of macaques with any of the three isolates resulted in high levels of primary plasma viremia by 1 week after inoculation. Viremia was quickly controlled following infection with SIVagm155; these animals have maintained CD4+ T-cell subsets and remain healthy. The plateau levels among SIVagm90- and SIVagm9063-inoculated macaques varied widely from 100 to 1 million copies/ml of plasma. Three of four animals from each of these groups progressed to AIDS. Setpoint viremia and the degree of CD4+-T-cell loss at 6 months postinfection were not significantly different between macaques inoculated with SIVagm90 and SIVagm9063. However these parameters were significantly different in SIVagm155 inoculated macaques (P values of <0.01). Considering all the macaques, the degree of CD4+-T-cell loss by 6 months postinfection correlated with the plateau levels of viremia. Thus, similar to SIVsm/mac infection of macaques and human AIDS, viral load is an excellent prognostic indicator of disease course. The inherent pathogenicity of natural SIVagm isolates varies, but such natural isolates are capable of inducing AIDS in macaques without prior macaque passage. PMID- 15795300 TI - A proline-rich region in the coxsackievirus 3A protein is required for the protein to inhibit endoplasmic reticulum-to-golgi transport. AB - The ability of the 3A protein of coxsackievirus B (CVB) to inhibit protein secretion was investigated for this study. Here we show that the ectopic expression of CVB 3A blocked the transport of both the glycoprotein of vesicular stomatitis virus, a membrane-bound secretory marker, and the alpha-1 protease inhibitor, a luminal secretory protein, at a step between the endoplasmic reticulum (ER) and the Golgi complex. CVB 3A contains a conserved proline-rich region in its N terminus. The importance of this proline-rich region was investigated by introducing Pro-to-Ala substitutions. The mutation of Pro19 completely abolished the ability of 3A to inhibit ER-to-Golgi transport. The mutation of Pro14, Pro17, or Pro20 also impaired this ability, but to a lesser extent. The mutation of Pro18 had no effect. We also investigated the possible importance of this proline-rich region for the function of 3A in viral RNA replication. To this end, we introduced the Pro-to-Ala mutations into an infectious cDNA clone of CVB3. The transfection of cells with in vitro transcribed RNAs of these clones gave rise to mutant viruses that replicated with wild-type characteristics. We concluded that the proline-rich region in CVB 3A is required for its ability to inhibit ER-to-Golgi transport, but not for its function in viral RNA replication. The functional relevance of the proline-rich region is discussed in light of the proposed structural model of 3A. PMID- 15795301 TI - Comparison of inflammatory and acute-phase responses in the brain and peripheral organs of the ME7 model of prion disease. AB - Chronic neurodegenerative diseases such as prion disease and Alzheimer's disease (AD) are reported to be associated with microglial activation and increased brain and serum cytokines and acute-phase proteins (APPs). Unlike AD, prion disease is also associated with a peripheral component in that the presumed causative agent, PrPSc, also accumulates in the spleen and other lymphoreticular organs. It is unclear whether the reported systemic acute-phase response represents a systemic inflammatory response to prion disease or merely reflects central nervous system (CNS) inflammation. For this study, we investigated whether intracerebrally initiated prion disease (ME7 model) provokes splenic, hepatic, or brain inflammatory and acute-phase responses. We detected no significant elevation of proinflammatory cytokines or activation of macrophages in the spleens of these animals, despite clear PrPSc deposition. Similarly, at 19 weeks we detected no significant elevation of transcripts for the APPs serum amyloid A, complement C3, pentraxin 3, and alpha2-antiplasmin in the liver, despite CNS neurodegeneration and splenic PrPSc deposition at this time. However, despite the low CNS expression levels of proinflammatory cytokines, there was robust expression of these APPs in degenerating brains. These findings suggest that PrPSc is not a stimulus for splenic macrophages and that neither peripheral PrPSc deposition nor CNS neurodegeneration is sufficient to produce a systemic acute-phase response. We also propose that serum cytokine and APP measurements are not useful during preclinical disease. Possible consequences of the clear chronic elevation of APPs in the CNS are discussed. PMID- 15795303 TI - In vivo evidence for instability of episomal human immunodeficiency virus type 1 cDNA. AB - Current regimens for the management of human immunodeficiency virus type 1 (HIV 1) infection suppress plasma viremia to below detectable levels for prolonged intervals. Nevertheless, there is a rapid resumption in plasma viremia if therapy is interrupted. Attempts to characterize the extent of viral replication under conditions of potent suppression and undetectable plasma viremia have been hampered by a lack of convenient assays that can distinguish latent from ongoing viral replication. Using episomal viral cDNA as a surrogate for ongoing replication, we previously presented evidence that viral replication persists in the majority of infected individuals with a sustained aviremic status. The labile nature of viral episomes and hence their validity as surrogate markers of ongoing replication in individuals with long-term-suppressed HIV-1 infection have been analyzed in short-term in vitro experiments with conflicting results. Since these in vitro experiments do not shed light on the long-term in vivo dynamics of episomal cDNA or recapitulate the natural targets of infection in vivo, we have analyzed the dynamics of episomal cDNA turnover in vivo by following the emergence of an M184V polymorphism in plasma viral RNA, in episomal cDNA, and in proviral DNA in patients on suboptimal therapies. We demonstrate that during acquisition of drug resistance, wild-type episomal cDNAs are replaced by M184V harboring episomes. Importantly, a complete replacement of wild-type episomes with M184V-containing episomes occurred while proviruses remained wild type. This indicates that episomal cDNAs are turned over by degradation rather than through death or tissue redistribution of the infected cell itself. Therefore, evolution of episomal viral cDNAs is a valid surrogate of ongoing viral replication in HIV 1-infected individuals. PMID- 15795304 TI - Weak palindromic consensus sequences are a common feature found at the integration target sites of many retroviruses. AB - Integration into the host genome is one of the hallmarks of the retroviral life cycle and is catalyzed by virus-encoded integrases. While integrase has strict sequence requirements for the viral DNA ends, target site sequences have been shown to be very diverse. We carefully examined a large number of integration target site sequences from several retroviruses, including human immunodeficiency virus type 1, simian immunodeficiency virus, murine leukemia virus, and avian sarcoma-leukosis virus, and found that a statistical palindromic consensus, centered on the virus-specific duplicated target site sequence, was a common feature at integration target sites for these retroviruses. PMID- 15795302 TI - A novel assay allows genotyping of the latent reservoir for human immunodeficiency virus type 1 in the resting CD4+ T cells of viremic patients. AB - A latent reservoir for human immunodeficiency virus type 1 (HIV-1) consisting of integrated provirus in resting memory CD4+ T cells prevents viral eradication in patients on highly active antiretroviral therapy (HAART). It is difficult to analyze the nature and dynamics of this reservoir in untreated patients and in patients failing therapy, because it is obscured by an excess of unintegrated viral DNA that constitutes the majority of viral species in resting CD4+ T cells from viremic patients. Therefore, we developed a novel culture assay that stimulates virus production from latent, integrated HIV-1 in resting CD4+ T cells in the presence of antiretroviral drugs that prevent the replication of unintegrated virus. Following activation, resting CD4+ T cells with integrated HIV-1 DNA produced virus particles for several days, with peak production at day 5. Using this assay, HIV-1 pol sequences from the resting CD4+ T cells of viremic patients were found to be genetically distinct from contemporaneous plasma virus. Despite the predominance of a relatively homogeneous population of drug-resistant viruses in the plasma of patients failing HAART, resting CD4+ T cells harbored a diverse array of wild-type and archival drug-resistant viruses that were less fit than plasma virus in the context of current therapy. These results provide the first direct evidence that resting CD4+ T cells serve as a stable reservoir for HIV-1 even in the setting of high levels of viremia. The ability to analyze archival species in viremic patients may have clinical utility in detecting drug resistant variants not present in the plasma. PMID- 15795305 TI - Induction of apoptosis by rewiring the signal transduction of Epstein-Barr virus oncoprotein LMP1 toward caspase activation. AB - The Epstein-Barr virus latent membrane protein 1 (LMP1) is an oncoprotein which mimics activated tumor necrosis factor receptor family members. Here we demonstrate the principle that an inducible association of the LMP1 cytoplasmic carboxyl terminus with caspase-8 by a heterodimerizing agent causes apoptosis. This process depends on the catalytic activity of caspase-8 and the ability of LMP1 to oligomerize constitutively at the plasma membrane. Our data indicate that chemical inducers of the association of the LMP1 carboxyl terminus with caspase-8 can kill LMP1-expressing cells selectively. Such compounds could be used as chemotherapeutic agents for LMP1-associated malignancies. PMID- 15795307 TI - Ex vivo stimulation of B cells latently infected with gammaherpesvirus 68 triggers reactivation from latency. AB - Murine gammaherpesvirus 68 (gammaHV68) infection of mice results in the establishment of a chronic infection, which is largely maintained through latent infection of B lymphocytes. Acute virus replication is almost entirely cleared by 2 weeks postinfection. Spontaneous reactivation of gammaHV68 from latently infected splenocytes upon ex vivo culture can readily be detected at the early stages of infection (e.g., day 16). However, by 6 weeks postinfection, very little spontaneous reactivation is detected upon explant into tissue culture. Here we report that stimulation of latently infected splenic B cells harvested at late times postinfection with cross-linking surface immunoglobulin (Ig), in conjunction with anti-CD40 antibody treatment, triggers virus reactivation. As expected, this treatment resulted in B-cell activation, as assessed by upregulation of CD69 on B cells, and ultimately B-cell proliferation. Since anti Ig/anti-CD40 stimulation resulted in splenic B-cell proliferation, we assessed whether this reactivation stimulus could overcome the previously characterized defect in virus reactivation of a v-cyclin null gammaHV68 mutant. This analysis demonstrated that anti-Ig/anti-CD40 stimulation could drive reactivation of the v cyclin null mutant virus in latently infected splenocytes, but not to the levels observed with wild-type gammaHV68. Thus, there appears to be a role for the v cyclin in B cells following anti-Ig/anti-CD40 stimulation independent of the induction of the cell cycle. Finally, to assess signals that are not mediated through the B-cell receptor, we demonstrate that addition of lipopolysaccharide to explanted splenocyte cultures also enhanced virus reactivation. These studies complement and extend previous analyses of Epstein-Barr virus and Kaposi's sarcoma-associated virus reactivation from latently infected cell lines by investigating reactivation of gammaHV68 from latently infected primary B cells recovered from infected hosts. PMID- 15795306 TI - Tissue-resident macrophages are productively infected ex vivo by primary X4 isolates of human immunodeficiency virus type 1. AB - Infection of macrophages has been implicated as a critical event in the transmission and persistence of human immunodeficiency virus type 1 (HIV-1). Here, we explore whether primary X4 HIV-1 isolates can productively infect tissue macrophages that have terminally differentiated in vivo. Using immunohistochemistry, HIV-1 RNA in situ hybridization, and confocal immunofluorescence microscopy, we demonstrate that macrophages residing in human tonsil blocks can be productively infected ex vivo by primary X4 HIV-1 isolates. This challenges the model in which macrophage tropism is a key determinant of the selective transmission of R5 HIV-1 strains. Infection of tissue macrophages by X4 HIV-1 may be highly relevant in vivo and contribute to key events in HIV-1 pathogenesis. PMID- 15795308 TI - Identification of a new quaternary neutralizing epitope on human immunodeficiency virus type 1 virus particles. AB - The selection of human monoclonal antibodies (MAbs) specific for human immunodeficiency virus (HIV) type 1 by binding assays may fail to identify Abs to quaternary epitopes on the intact virions. The HIV neutralization assay was used for the selection of human MAb 2909, which potently neutralizes SF162 and recognizes an epitope on the virus surface but not on soluble proteins. Three regions of gp120, the V2 and V3 loops and the CD4 binding domain, contribute to the epitope recognized by MAb 2909. The existence of such a unique MAb, which defines a complex epitope formed by a quaternary structure, suggests that there may be other new neutralizing HIV epitopes to target with vaccines. PMID- 15795309 TI - Evidence of serological cross-reactivity between genotype 1 and genotype 3 erythrovirus infections. PMID- 15795310 TI - Secondary structure and gating rearrangements of transmembrane segments in rat P2X4 receptor channels. AB - P2X receptors are cation selective channels that are activated by extracellular nucleotides. These channels are likely formed by three identical or related subunits, each having two transmembrane segments (TM1 and TM2). To identify regions that undergo rearrangement during gating and to probe their secondary structure, we performed tryptophan scanning mutagenesis on the two putative TMs of the rat P2X4 receptor channel. Mutant channels were expressed in Xenopus oocytes, concentration-response relationships constructed for ATP, and the EC50 estimated by fitting the Hill equation to the data. Of the 22 mutations in TM1 and 24 in TM2, all but one in TM1 and seven in TM2 result in functional channels. Interestingly, the majority of the functional mutants display an increased sensitivity to ATP, and in general these perturbations are more pronounced for TM2 when compared with TM1. For TM1 and for the outer half of TM2, the perturbations are consistent with these regions adopting alpha-helical secondary structures. In addition, the greatest perturbations in the gating equilibrium occur for mutations near the outer ends of both TM1 and TM2. Surface biotinylation experiments reveal that all the nonfunctional mutants traffic to the surface membrane at levels comparable to the WT channel, suggesting that these mutations likely disrupt ion conduction or gating. Taken together, these results suggest that the outer parts of TM1 and TM2 are helical and that they move during activation. The observation that the majority of nonconducting mutations are clustered toward the inner end of TM2 suggests a critical functional role for this region. PMID- 15795313 TI - The NIH policy on enhancing public access to publications resulting from NIH funded research: can we streamline the process for our authors? PMID- 15795311 TI - Mechanism of the voltage sensitivity of IRK1 inward-rectifier K+ channel block by the polyamine spermine. AB - IRK1 (Kir2.1) inward-rectifier K+ channels exhibit exceedingly steep rectification, which reflects strong voltage dependence of channel block by intracellular cations such as the polyamine spermine. On the basis of studies of IRK1 block by various amine blockers, it was proposed that the observed voltage dependence (valence approximately 5) of IRK1 block by spermine results primarily from K+ ions, not spermine itself, traversing the transmembrane electrical field that drops mostly across the narrow ion selectivity filter, as spermine and K+ ions displace one another during channel block and unblock. If indeed spermine itself only rarely penetrates deep into the ion selectivity filter, then a long blocker with head groups much wider than the selectivity filter should exhibit comparably strong voltage dependence. We confirm here that channel block by two molecules of comparable length, decane-bis-trimethylammonium (bis-QA(C10)) and spermine, exhibit practically identical overall voltage dependence even though the head groups of the former are much wider ( approximately 6 A) than the ion selectivity filter ( approximately 3 A). For both blockers, the overall equilibrium dissociation constant differs from the ratio of apparent rate constants of channel unblock and block. Also, although steady-state IRK1 block by both cations is strongly voltage dependent, their apparent channel-blocking rate constant exhibits minimal voltage dependence, which suggests that the pore becomes blocked as soon as the blocker encounters the innermost K+ ion. These findings strongly suggest the existence of at least two (potentially identifiable) sequentially related blocked states with increasing numbers of K+ ions displaced. Consequently, the steady-state voltage dependence of IRK1 block by spermine or bis-QA(C10) should increase with membrane depolarization, a prediction indeed observed. Further kinetic analysis identifies two blocked states, and shows that most of the observed steady-state voltage dependence is associated with the transition between blocked states, consistent with the view that the mutual displacement of blocker and K+ ions must occur mainly as the blocker travels along the long inner pore. PMID- 15795312 TI - Type-3 ryanodine receptors mediate hypoxia-, but not neurotransmitter-induced calcium release and contraction in pulmonary artery smooth muscle cells. AB - In this study we examined the expression of RyR subtypes and the role of RyRs in neurotransmitter- and hypoxia-induced Ca2+ release and contraction in pulmonary artery smooth muscle cells (PASMCs). Under perforated patch clamp conditions, maximal activation of RyRs with caffeine or inositol triphosphate receptors (IP3Rs) with noradrenaline induced equivalent increases in [Ca2+]i and Ca2+ activated Cl- currents in freshly isolated rat PASMCs. Following maximal IP3 induced Ca2+ release, neither caffeine nor chloro-m-cresol induced a response, whereas prior application of caffeine or chloro-m-cresol blocked IP3-induced Ca2+ release. In cultured human PASMCs, which lack functional expression of RyRs, caffeine failed to affect ATP-induced increases in [Ca2+]i in the presence and absence of extracellular Ca2+. The RyR antagonists ruthenium red, ryanodine, tetracaine, and dantrolene greatly inhibited submaximal noradrenaline- and hypoxia-induced Ca2+ release and contraction in freshly isolated rat PASMCs, but did not affect ATP-induced Ca2+ release in cultured human PASMCs. Real-time quantitative RT-PCR and immunofluorescence staining indicated similar expression of all three RyR subtypes (RyR1, RyR2, and RyR3) in freshly isolated rat PASMCs. In freshly isolated PASMCs from RyR3 knockout (RyR3-/-) mice, hypoxia-induced, but not submaximal noradrenaline-induced, Ca2+ release and contraction were significantly reduced. Ruthenium red and tetracaine can further inhibit hypoxic increase in [Ca2+]i in RyR3-/- mouse PASMCs. Collectively, our data suggest that (a) RyRs play an important role in submaximal noradrenaline- and hypoxia-induced Ca2+ release and contraction; (b) all three subtype RyRs are expressed; and (c) RyR3 gene knockout significantly inhibits hypoxia-, but not submaximal noradrenaline-induced Ca2+ and contractile responses in PASMCs. PMID- 15795314 TI - The yeast S phase checkpoint enables replicating chromosomes to bi-orient and restrain spindle extension during S phase distress. AB - The budding yeast S phase checkpoint responds to hydroxyurea-induced nucleotide depletion by preventing replication fork collapse and the segregation of unreplicated chromosomes. Although the block to chromosome segregation has been thought to occur by inhibiting anaphase, we show checkpoint-defective rad53 mutants undergo cycles of spindle extension and collapse after hydroxyurea treatment that are distinct from anaphase cells. Furthermore, chromatid cohesion, whose dissolution triggers anaphase, is dispensable for S phase checkpoint arrest. Kinetochore-spindle attachments are required to prevent spindle extension during replication blocks, and chromosomes with two centromeres or an origin of replication juxtaposed to a centromere rescue the rad53 checkpoint defect. These observations suggest that checkpoint signaling is required to generate an inward force involved in maintaining preanaphase spindle integrity during DNA replication distress. We propose that by promoting replication fork integrity under these conditions Rad53 ensures centromere duplication. Replicating chromosomes can then bi-orient in a cohesin-independent manner to restrain untimely spindle extension. PMID- 15795316 TI - ArfGAP1 dynamics and its role in COPI coat assembly on Golgi membranes of living cells. AB - Secretory protein trafficking relies on the COPI coat, which by assembling into a lattice on Golgi membranes concentrates cargo at specific sites and deforms the membranes at these sites into coated buds and carriers. The GTPase-activating protein (GAP) responsible for catalyzing Arf1 GTP hydrolysis is an important part of this system, but the mechanism whereby ArfGAP is recruited to the coat, its stability within the coat, and its role in maintenance of the coat are unclear. Here, we use FRAP to monitor the membrane turnover of GFP-tagged versions of ArfGAP1, Arf1, and coatomer in living cells. ArfGAP1 underwent fast cytosol/Golgi exchange with approximately 40% of the exchange dependent on engagement of ArfGAP1 with coatomer and Arf1, and affected by secretory cargo load. Permanent activation of Arf1 resulted in ArfGAP1 being trapped on the Golgi in a coatomer dependent manner. These data suggest that ArfGAP1, coatomer and Arf1 play interdependent roles in the assembly-disassembly cycle of the COPI coat in vivo. PMID- 15795315 TI - A systems analysis of importin-{alpha}-{beta} mediated nuclear protein import. AB - Importin-beta (Impbeta) is a major transport receptor for Ran-dependent import of nuclear cargo. Impbeta can bind cargo directly or through an adaptor such as Importin-alpha (Impalpha). Factors involved in nuclear transport have been well studied, but systems analysis can offer further insight into regulatory mechanisms. We used computer simulation and real-time assays in intact cells to examine Impalpha-beta-mediated import. The model reflects experimentally determined rates for cargo import and correctly predicts that import is limited principally by Impalpha and Ran, but is also sensitive to NTF2. The model predicts that CAS is not limiting for the initial rate of cargo import and, surprisingly, that increased concentrations of Impbeta and the exchange factor, RCC1, actually inhibit rather than stimulate import. These unexpected predictions were all validated experimentally. The model revealed that inhibition by RCC1 is caused by sequestration of nuclear Ran. Inhibition by Impbeta results from depletion nuclear RanGTP, and, in support of this mechanism, expression of mRFP Ran reversed the inhibition. PMID- 15795317 TI - The extracellular domains of FasL and Fas are sufficient for the formation of supramolecular FasL-Fas clusters of high stability. AB - Using fluorescent variants of Fas and FasL, we show that membrane FasL and Fas form supramolecular clusters that are of flexible shape, but nevertheless stable and persistent. Membrane FasL-induced Fas clusters were formed in caspase-8- or FADD-deficient cells or when a cytoplasmic deletion mutant of Fas was used suggesting that cluster formation is independent of the assembly of the cytoplasmic Fas signaling complex and downstream activated signaling pathways. In contrast, cross-linked soluble FasL failed to aggregate the cytoplasmic deletion mutant of Fas, but still induced aggregation of signaling competent full-length Fas. Moreover, membrane FasL-induced Fas cluster formation occurred in the presence of the lipid raft destabilizing component methyl-beta-cyclodextrin, whereas Fas aggregation by soluble FasL was blocked. Together, these data suggest that the extracellular domains of Fas and FasL alone are sufficient to drive membrane FasL-induced formation of supramolecular Fas-FasL complexes, whereas soluble FasL-induced Fas aggregation is dependent on lipid rafts and mechanisms associated with the intracellular domain of Fas. PMID- 15795318 TI - {beta}1 Integrin and IL-3R coordinately regulate STAT5 activation and anchorage dependent proliferation. AB - We previously demonstrated that integrin-dependent adhesion activates STAT5A, a well known target of IL-3-mediated signaling. Here, we show that in endothelial cells the active beta1 integrin constitutively associates with the unphosphorylated IL-3 receptor (IL-3R) beta common subunit. This association is not sufficient for activating downstream signals. Indeed, only upon fibronectin adhesion is Janus Kinase 2 (JAK2) recruited to the beta1 integrin-IL-3R complex and triggers IL-3R beta common phosphorylation, leading to the formation of docking sites for activated STAT5A. These events are IL-3 independent but require the integrity of the IL-3R beta common. IL-3 treatment increases JAK2 activation and STAT5A and STAT5B tyrosine and serine phosphorylation and leads to cell cycle progression in adherent cells. Expression of an inactive STAT5A inhibits cell cycle progression upon IL-3 treatment, identifying integrin-dependent STAT5A activation as a priming event for IL-3-mediated S phase entry. Consistently, overexpression of a constitutive active STAT5A leads to anchorage-independent cell cycle progression. Therefore, these data provide strong evidence that integrin-dependent STAT5A activation controls IL-3-mediated proliferation. PMID- 15795319 TI - Three-dimensional EM structure of the ectodomain of integrin {alpha}V{beta}3 in a complex with fibronectin. AB - Integrins are alphabeta heterodimeric cell surface receptors that mediate transmembrane signaling by binding extracellular and cytoplasmic ligands. The ectodomain of integrin alphaVbeta3 crystallizes in a bent, genuflexed conformation considered to be inactive (unable to bind physiological ligands in solution) unless it is fully extended by activating stimuli. We generated a stable, soluble complex of the Mn(2+)-bound alphaVbeta3 ectodomain with a fragment of fibronectin (FN) containing type III domains 7 to 10 and the EDB domain (FN7-EDB-10). Transmission electron microscopy and single particle image analysis were used to determine the three-dimensional structure of this complex. Most alphaVbeta3 particles, whether unliganded or FN-bound, displayed compact, triangular shapes. A difference map comparing ligand-free and FN-bound alphaVbeta3 revealed density that could accommodate the RGD-containing FN10 in proximity to the ligand-binding site of beta3, with FN9 just adjacent to the synergy site binding region of alphaV. We conclude that the ectodomain of alphaVbeta3 manifests a bent conformation that is capable of stably binding a physiological ligand in solution. PMID- 15795320 TI - Cellular and pharmacogenetics foundation of synergistic interaction of pemetrexed and gemcitabine in human non-small-cell lung cancer cells. AB - Gemcitabine and pemetrexed are effective agents in the treatment of non-small cell lung cancer (NSCLC), and the present study investigates cellular and genetic aspects of their interaction against A549, Calu-1, and Calu-6 cells. Cells were treated with pemetrexed and gemcitabine, and their interaction was assessed using the combination index. The role of drug metabolism in gemcitabine cytotoxicity was examined with inhibitors of deoxycytidine kinase (dCK), 5'-nucleotidase, and cytidine deaminase, whereas the role of pemetrexed targets, thymidylate synthase (TS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT) in drug chemosensitivity was analyzed in cytotoxicity rescue studies. The effect of gemcitabine and pemetrexed on Akt phosphorylation was investigated with enzyme-linked immunosorbent assay, whereas quantitative polymerase chain reaction (PCR) was used to study target gene-expression profiles and its modulation by each drug. Synergistic cytotoxicity was demonstrated, and pemetrexed significantly decreased the amount of phosphorylated Akt, enhanced apoptosis, and increased the expression of dCK in A549 and Calu-6 cells, as well as the expression of the human nucleoside equilibrative transporter 1 (hENT1) in all cell lines. PCR demonstrated a correlation between dCK expression and gemcitabine sensitivity, whereas expression of TS, DHFR, and GARFT was predictive of pemetrexed chemosensitivity. These data demonstrated that 1) gemcitabine and pemetrexed synergistically interact against NSCLC cells through the suppression of Akt phosphorylation and induction of apoptosis; 2) the gene expression profile of critical genes may predict for drug chemosensitivity; and 3) pemetrexed enhances dCK and hENT1 expression, thus suggesting the role of gene-expression modulation for rational development of chemotherapy combinations. PMID- 15795321 TI - Akt is essential for insulin modulation of amphetamine-induced human dopamine transporter cell-surface redistribution. AB - Uptake by the dopamine transporter (DAT) is the primary pathway for the clearance of extracellular dopamine (DA) and consequently for regulating the magnitude and duration of dopaminergic signaling. Amphetamine (AMPH) has been shown to decrease simultaneously DAT cell-surface expression and [(3)H]DA uptake. We have shown that insulin and its subsequent signaling through the phosphatidylinositol 3 kinase (PI3K)-dependent pathway oppose this effect of AMPH by promoting increased cell-surface expression. Here, we used human embryonic kidney 293 cells stably expressing the human DAT (hDAT cells) to investigate the downstream cellular components important for this effect of insulin. Akt is a protein kinase effector immediately downstream of PI3K. Both overexpression of a dominant-negative mutant of Akt (K179R) and the addition of 1-(5-chloronaphthalene-1-sulfonyl)-1H hexahydro-1,4-diazepine HCl (ML9), a pharmacological inhibitor of Akt, decreased cell-surface expression of DAT, suggesting a role of basal Akt signaling in the homoeostasis of DAT. Moreover, expression of a constitutively active Akt mutant reduced the ability of AMPH to decrease hDAT cell-surface expression as well as [(3)H]DA uptake. In contrast, overexpression of K179R blocked the ability of insulin to oppose AMPH-induced reduction of hDAT cell-surface expression and [(3)H]DA uptake, as did ML9. Our data demonstrate that hDAT cell-surface expression is regulated by the insulin signaling pathway and that Akt plays a key role in the hormonal modulation of AMPH-induced hDAT trafficking and in the regulation of basal hDAT cell-surface expression. PMID- 15795322 TI - AFos dissociates cardiac myocyte hypertrophy and expression of the pathological gene program. AB - BACKGROUND: Although induction of activator protein-1 (AP-1) transcription factor activity has been observed in cardiac hypertrophy, a direct role for AP-1 in myocardial growth and gene expression remains obscure. METHODS AND RESULTS: Hypertrophy was induced in cultured neonatal rat cardiomyocytes with phenylephrine or overexpression of a constitutively active MAP3K, MKK6. In both treatment groups, induction of the pathological gene profile was observed, ie, expression of beta-myosin heavy chain (betaMHC), atrial/brain natriuretic peptides (ANP/BNP), and skeletal alpha-actin (sACT) was increased, whereas expression for alpha-myosin heavy chain (alphaMHC) and the sarcoplasmic reticulum Ca2+-ATPase (SERCA) genes was repressed. The role of AP-1 in the hypertrophic phenotype was evaluated with the use of an adenoviral construct expressing a dominant negative mutant of the c-Fos proto-oncogene (AdAFos). Although AFos did not change the myocyte growth response, it abrogated the gene profile to both agonists, including the upregulation of both alphaMHC and SERCA expression. CONCLUSIONS: Although c-Fos/AP-1 is necessary for induction of the pathological/fetal gene program, it does not appear to be critical for cardiomyocyte hypertrophy. PMID- 15795323 TI - Pivotal role of gp91phox-containing NADH oxidase in lipopolysaccharide-induced tumor necrosis factor-alpha expression and myocardial depression. AB - BACKGROUND: Lipopolysaccharide (LPS) induces cardiomyocyte tumor necrosis factor alpha (TNF-alpha) production, which is responsible for myocardial depression during sepsis. The aim of this study was to investigate the role of gp91phox containing NADH oxidase signaling in cardiomyocyte TNF-alpha expression and myocardial dysfunction induced by LPS. METHODS AND RESULTS: In cultured mouse neonatal cardiomyocytes, LPS increased NADH oxidase (gp91phox subunit) expression and superoxide generation. Deficiency of gp91phox or inhibition of NADH oxidase blocked TNF-alpha expression stimulated by LPS. TNF-alpha induction was also inhibited by tempol, N-acetylcysteine, or 1,3-dimethyl-2-thiourea. NADH oxidase activation by LPS increased ERK1/2 and p38 phosphorylation, and inhibition of ERK1/2 and p38 phosphorylation blocked the effect of NADH oxidase on TNF-alpha expression. Isolated mouse hearts were perfused with LPS (5 microg/mL) alone or in the presence of apocynin for 1 hour. Myocardial TNF-alpha production was decreased in gp91phox-deficient or apocynin-treated hearts compared with those of wild type (P<0.05). To investigate the role of gp91phox-containing NADH oxidase in endotoxemia, mice were treated with LPS (4 mg/kg IP) for 4 and 24 hours, and their heart function was measured with a Langendorff system. Deficiency of gp91phox significantly attenuated LPS-induced myocardial depression (P<0.05). CONCLUSIONS: gp91phox-Containing NADH oxidase is pivotal in LPS-induced TNF-alpha expression and cardiac depression. Effects of NADH oxidase activation are mediated by ERK1/2 and p38 MAPK pathway. The present results suggest that gp91phox-containing NADH oxidase may represent a potential therapeutic target for myocardial dysfunction in sepsis. PMID- 15795324 TI - Evidence of human thrombomodulin domain as a novel angiogenic factor. AB - BACKGROUND: Thrombomodulin is an anticoagulant, endothelial-cell-membrane glycoprotein. A recombinant thrombomodulin domain containing 6 epidermal growth factor-like structures exhibits mitogenic activity. This study explored the novel angiogenic effects of the recombinant domain using in vitro and in vivo models. METHODS AND RESULTS: Human recombinant thrombomodulin containing 6 epidermal growth factor-like structures (TMD2) and TMD2 plus a serine and threonine-rich domain (TMD23) were prepared using the Pichia pastoris expression system. Combined with purified TMD2 or TMD23, thrombin effectively activated protein C. TMD23 had higher activity than TMD2 in stimulating DNA synthesis in cultured human umbilical vein endothelial cells. Additionally, TMD23 stimulated chemotactic motility and capillarylike tube formation in human umbilical vein endothelial cells, an effect mediated through phosphorylation of extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase and the phosphatidylinositol-3 kinase/Akt/endothelial nitric oxide synthase pathway. TMD23 also stimulated endothelial cell expression of matrix metalloproteinases and plasminogen activators, which mediated extracellular proteolysis, leading to endothelial cell invasion and migration during angiogenesis. Furthermore, TMD23 containing implants in rat cornea induced ingrowth of new blood vessels from the limbus. With the murine angiogenesis assay, TMD23 not only induced neovascularization coinjected with Matrigel and heparin but also enhanced angiogenesis in Matrigel containing melanoma A2058 cells in nude mice. CONCLUSIONS: The recombinant thrombomodulin domain TMD23 enhanced the angiogenic response in vitro and in vivo, suggesting that thrombomodulin fragments may play a role in the formation of new vessels. These findings may provide a new therapeutic option for treating ischemic diseases. PMID- 15795325 TI - Thrombosis modulates arterial drug distribution for drug-eluting stents. AB - BACKGROUND: Drug-eluting stents deliver potent compounds directly to arterial segments but can become clot laden when deployed. The question arises as to whether thrombi affect drug elution and arterial uptake. METHODS AND RESULTS: Paclitaxel transport and retention were assessed in clots of different blood components. Diffusivity, affected by clot organization, is fastest in fibrin (approximately 347 microm2/s), slower in fibrin-red blood cell clots (34.98 microm2/s), and slowest in whole-blood clots (3.55 microm2/s). Blood cells bind and retain paclitaxel such that levels in clot increase linearly with red cell fraction. At physiological hematocrit, clot retains 3 times the amount of paclitaxel in surrounding solutions. Computational models predict that the potential of thrombus to absorb, retain, and release drug or to act as a barrier to drug delivery depends on clot geometry and strut position in clot relative to the vessel wall. Clot between artery and stent can reduce uptake 10-fold, whereas clot overlying the stent can shield drug from washout, increasing uptake. Model assumptions were confirmed and predictions were validated in a novel rat model that introduces thrombosis within stented aortas where nonocclusive thrombus acts as capacitive space for drug and shifts drug levels to decrease tissue uptake 2 fold. CONCLUSIONS: Thrombus apposed on stents creates large variations in drug uptake and can act to either increase or decrease wall deposition according to the clot and stent geometry. Arterial deposition of drug from stents deployed in clots will be highly variable and unpredictable unless the clot can be adequately controlled or removed. PMID- 15795326 TI - Celecoxib decreases endothelial tissue factor expression through inhibition of c Jun terminal NH2 kinase phosphorylation. AB - BACKGROUND: Despite potential antiinflammatory properties, the use of selective cyclooxygenase-2 inhibitors (coxibs) in patients with cardiovascular diseases has been questioned because of a possibly increased thrombotic risk. Tissue factor (TF), a key protein for initiation of coagulation, has been implicated in the pathogenesis of atherosclerosis and thrombosis. Hence, we examined the effect of different coxibs on TF expression. METHODS AND RESULTS: Celecoxib (10(-5) mol/L), but not rofecoxib (10(-7) to 10(-5) mol/L) or the experimental coxib NS-398 (10( 7) to 10(-5) mol/L), decreased tumor necrosis factor-alpha-induced TF expression and activity in human aortic endothelial cells. Celecoxib (10(-5) mol/L) reduced activation of c-jun terminal NH2 kinase (JNK), whereas it did not affect p38 mitogen-activated protein (MAP) kinase or p44/42 MAP kinase; in contrast, JNK activation was not affected by rofecoxib (10(-5) mol/L) or NS-398 (10(-5) mol/L). TF expression was reduced in a concentration-dependent manner by pretreatment with SP600125 (10(-7) to 10(-6) mol/L), a specific inhibitor of JNK, which confirms that JNK regulates tumor necrosis factor-alpha-induced TF expression. CONCLUSIONS: Celecoxib reduced TF expression and activity in human aortic endothelial cells. Because neither rofecoxib nor the experimental coxib NS-398 affected TF expression, this effect occurs independently of COX-2 inhibition; it is rather mediated through inhibition of JNK phosphorylation. These data indicate a distinct heterogeneity within this class of drugs, which may be clinically relevant, especially for patients with atherosclerotic vascular diseases. PMID- 15795327 TI - 25-hydroxyvitamin D3-1alpha-hydroxylase is expressed in human vascular smooth muscle cells and is upregulated by parathyroid hormone and estrogenic compounds. AB - BACKGROUND: 1,25(OH)2 vitamin D3 exerts multiple effects in human vascular smooth muscle cells (VSMCs). We therefore tested the possibility that VSMCs possess an endogenous 25-hydroxyvitamin D3-1alpha-hydroxylase system, the final enzyme in the biosynthetic pathway of 1,25(OH)2D3. METHODS AND RESULTS: We assessed the expression and activity of 25-hydroxyvitamin D3-1alpha-hydroxylase by real-time polymerase chain reaction and the conversion of 25(OH)D3 into 1,25(OH)2D3. First, 25-hydroxyvitamin D3-1alpha-hydroxylase mRNA was identified in cultured VSMCs by real-time polymerase chain reaction. Second, in cells treated daily (3 days) with parathyroid hormone (66 nmol/L), estradiol-17beta (30 nmol/L), raloxifene (3 micromol/L), and the phytoestrogens genistein (3 micromol/L), biochainin A (3 micromol/L), and 6-carboxy biochainin A (30 nmol/L), 25-hydroxyvitamin D3-1alpha hydroxylase mRNA increased by 43+/-13%, (P<0.05) 7+/-24% (P=NS), 176+/-28% (P<0.01), 65+/-11% (P<0.05), 152+/-24% (P<0.01), and 71+/-9% (P<0.05), respectively. Third, production of 1,25(OH)2D3 from 25(OH)D3 was seen with a Km of 25 ng/mL and increased dose dependently after treatment with parathyroid hormone, genistein, and the phytosetrogen derivative 6-carboxy biochainin A. Estradiol-17beta and biochainin A also increased the generation of 1,25(OH)2D3 by 40+/-23% (P<0.05) and 55+/-13% (P<0.05), respectively. CONCLUSIONS: We provide here the first evidence for the expression of an enzymatically active 25(OH)D3 1alpha-hydroxylase system in human VSMCs, which can be upregulated by parathyroid hormone and estrogenic compounds. Because exogenous vitamin D inhibits VSMC proliferation, the role of this system as an autocrine mechanism to curb changes in VSMC proliferation and phenotype is a subject for future investigation. PMID- 15795328 TI - Serum and glucocorticoid-responsive kinase-1 regulates cardiomyocyte survival and hypertrophic response. AB - BACKGROUND: Serum- and glucocorticoid-responsive kinase-1 (SGK1), a serine threonine kinase that is highly expressed in the heart, has been previously reported to regulate sodium channels. Because SGK1 is a PI 3-kinase-dependent kinase with structural homology to Akt, we examined its regulation in the heart and its effects on cardiomyocyte (CM) apoptosis and hypertrophy in vitro. METHODS AND RESULTS: Rats were subjected to aortic banding, and expression of total and phosphorylated SGK1 was examined. Both phospho- and total SGK1 increased 2 to 7 days after banding. Phospho-SGK1 was also upregulated in CMs stimulated in vitro with IGF-I or phenylephrine. Infection of CMs with an adenoviral vector encoding constitutively active SGK1 (Ad.SGK1.CA) inhibited apoptosis after serum deprivation or hypoxia (P<0.05), whereas expression of kinase-dead SGK1 (Ad.SGK1.KD) increased it and partially mitigated the protective effects of IGF-I (P<0.05). SGK1 activation was also sufficient to increase cell size, protein synthesis, sarcomere organization, and ANF expression both at baseline and in response to phenylephrine but was not necessary for the hypertrophic response to phenylephrine. Evaluation of potential downstream signaling pathways demonstrated that SGK1 induces phosphorylation of tuberin, p70s6kinase, and GSK3beta in CMs, which may contribute to its effects. CONCLUSIONS: SGK1 is dynamically regulated during acute biomechanical stress in the heart and inhibits CM apoptosis while enhancing the hypertrophic response. PMID- 15795329 TI - Effect of hydrodynamics-based gene delivery of plasmid DNA encoding interleukin-1 receptor antagonist-Ig for treatment of rat autoimmune myocarditis: possible mechanism for lymphocytes and noncardiac cells. AB - BACKGROUND: Interleukin-1 (IL-1) is a powerful and important cytokine in myocarditis. The purpose of this study was to evaluate the effect and possible mechanism of hydrodynamics-based delivery of the IL-1 receptor antagonist (IL 1RA)-immunoglobulin (Ig) gene for treatment of rat experimental autoimmune myocarditis (EAM). METHODS AND RESULTS: On the day after immunization, rats were transfected with either pCAGGS encoding IL-1RA-Ig or pCAGGS encoding Ig alone. On day 17, IL-1RA-Ig gene therapy was effective in controlling EAM, as monitored by a decreased ratio of heart weight to body weight, reduced myocarditis areas, reduced gene expression of atrial natriuretic peptide in hearts, and improved cardiac function in echocardiographic and hemodynamic parameters. Examination of the expression of IL-1-related genes in purified cells from EAM hearts suggested that ectopic IL-1RA-Ig-acting target cells were alphabetaT cells and noncardiomyocytic noninflammatory cells such as fibroblasts, smooth muscle cells, and endothelial cells. Therefore, we examined the effect of serum containing IL 1RA-Ig on the expression of immune-relevant genes within noncardiomyocytic cells cultured from EAM hearts or concanavalin A-stimulated lymphocytes derived from lymph nodes in EAM-affected rats. The expression of immunologic molecules (prostaglandin E synthase, cyclooxygenase-2, and IL-1beta) in cultivated noncardiomyocytic cells and Th1 cytokines (IL-2 and IFN-gamma) in lymphocytes was significantly decreased by the serum containing IL-1RA-Ig. CONCLUSIONS: EAM was suppressed by hydrodynamics-based delivery of plasmid DNA encoding IL-1RA-Ig. In addition, IL-1RA-Ig suppressed gene expression of prostaglandin synthases and IL 1 in noncardiomyocytic cells and Th1 cytokines in lymphocytes. PMID- 15795330 TI - Targeting 2A protease by RNA interference attenuates coxsackieviral cytopathogenicity and promotes survival in highly susceptible mice. AB - BACKGROUND: Enteroviridae such as coxsackievirus B3 (CVB3) are important infectious agents involved in viral heart disease, hepatitis, and pancreatitis, but no specific antiviral therapy is available. METHODS AND RESULTS: The aim of the present study was to evaluate the impact of RNA interference on viral replication, cytopathogenicity, and survival. Small interfering RNA (siRNA) molecules were designed against the viral 2A region (siRNA-2A), which is considered to be highly conserved and essential for both virus maturation and host cytopathogenicity. siRNA-2A exhibited a significant protective effect on cell viability mediated by marked inhibition of CVB3 gene expression and viral replication. In highly susceptible type I interferon receptor-knockout mice, siRNA-2A led to significant reduction of viral tissue titers, attenuated tissue damage, and prolonged survival. Repeated siRNA-2A transfection was associated with a further improvement of survival. Various control siRNA molecules had no protective effect in vitro or in vivo. CONCLUSIONS: RNA interference directed against the 2A protease encoding genomic region effectively confers intracellular immunity toward CVB3-mediated cell injury and improves survival, suggesting a potential role for RNA interference for future treatment options targeting enteroviral diseases. PMID- 15795332 TI - Endothelial dysfunction in childhood infection. AB - BACKGROUND: Atherosclerosis begins in early life, and endothelial dysfunction is recognized as a key initiating event in the development of atherosclerosis. Although infection has been implicated in endothelial dysfunction and atherogenesis, the impact of acute common childhood infections on the vascular endothelium is unknown. METHODS AND RESULTS: We studied 600 children aged 10 years drawn from the Avon Longitudinal Study of Parents and Children. The children were divided into 3 groups: those with current acute infection (AI; n=135; 73 boys and 62 girls); a convalescent group with infection in the past 2 weeks (n=166; 78 boys and 88 girls), and a healthy control group (n=299; 131 boys and 168 girls). Endothelial function was determined in all subjects by high resolution ultrasound to measure brachial artery flow-mediated dilation (FMD) and was expressed as the percentage change in diameter from baseline after reactive hyperemia. FMD was repeated in 40 children in the AI group and 50 in the control group after a mean interval of 1 year. FMD was lower in both the AI group (6.3+/ 2.7%, mean+/-SD) and the convalescent group (8.1+/-3.1%) than in the control group (9.7+/-2.5%; P<0.001 for both). The observed differences in FMD remained after adjustment for potential confounding variables. At the repeat visit, FMD was unchanged in controls (P=0.85) but improved in the AI group (P<0.001). CONCLUSIONS: Acute infection in childhood is associated with impaired endothelium dependent vasodilation. These findings support a potential role for previously unsuspected extrinsic inflammatory stimuli in the pathogenesis of early atherosclerosis. PMID- 15795331 TI - Ablation of MEK kinase 1 suppresses intimal hyperplasia by impairing smooth muscle cell migration and urokinase plasminogen activator expression in a mouse blood-flow cessation model. AB - BACKGROUND: Migration, proliferation, and matrix-degrading protease expression of smooth muscle cells (SMCs) are major features of intimal hyperplasia after vascular injury. Although MEK kinase 1 (MEKK1) has been shown to regulate cell migration and urokinase plasminogen activator (uPA) expression, the precise role of MEKK1 in this process remains unknown. METHODS AND RESULTS: We triggered a vascular remodeling model by complete ligation of the right common carotid artery in wild-type (WT) and MEKK1-null (MEKK1-/-) mice. The intimal areas 28 days after ligation were significantly decreased in the ligated MEKK1-/- arteries compared with WT arteries (28+/-8 versus 65+/-17 microm2, P<0.05). There were no differences in the ratios of proliferating cell nuclear antigen (PCNA)-positive cells to total cells within the arterial wall between WT and MEKK1-/- arteries. Proliferation capacity also did not differ between WT and MEKK1-/- cultured aortic smooth muscle cells (AoSMCs). In contrast, the number of intimal PCNA positive cells 7 days after ligation was significantly smaller in MEKK1-/- arteries. Three different migration assays revealed that migration and invasion of MEKK1-/- AoSMCs were markedly impaired. Addition of full-length MEKK1 restored the migration capacity of MEKK1-/- AoSMCs. The number of MEKK1-/- AoSMCs showing lamellipodia formation by epithelial growth factor was significantly smaller compared with those of WT SMCs. Furthermore, uPA expression after ligation was markedly decreased in MEKK1-/- arteries. CONCLUSIONS: MEKK1 is implicated in vascular remodeling after blood-flow cessation by regulating the migration and uPA expression of SMCs. MEKK1 is a potential target for drug development to prevent vascular remodeling. PMID- 15795333 TI - CD40/CD40 ligand signaling in mouse cerebral microvasculature after focal ischemia/reperfusion. AB - BACKGROUND: CD40/CD40 ligand (CD40L) signaling contributes to proinflammatory and prothrombogenic responses in the vasculature. CD40/CD40L expression is elevated in patients after a transient ischemic attack or stroke. The purpose of this study was to investigate the role of CD40/CD40L signaling in cerebral microvascular dysfunction and tissue injury response to middle cerebral artery occlusion (MCAO) and reperfusion. METHODS AND RESULTS: Intravital fluorescence microscopy was used to visualize the cerebral microcirculation of wild-type (WT), CD40-deficient, and CD40L-deficient mice subjected to 1-hour MCAO and 4-hour reperfusion. The adhesion of platelets and of leukocytes and vascular permeability were measured in postcapillary venules after 4-hour and 1-hour reperfusions, respectively. Cerebral infarct volume was analyzed 24 hours after reperfusion. Platelet and leukocyte adhesion was elevated and blood/brain barrier function was compromised by MCAO in WT mice. Blood cell recruitment and increased permeability were blunted in both CD40-deficient and CD40L-deficient mice. Infarct volume was also reduced in CD40- and CD40L-deficient mice compared with WT mice. CONCLUSIONS: Our findings indicate that CD40/CD40L signaling contributes to inflammatory and prothrombogenic responses and brain infarction induced by MCAO and reperfusion. The CD40/CD40L dyad may play a significant pathogenic role in the acute phase of ischemic stroke. PMID- 15795334 TI - What if the patient were your mother? PMID- 15795335 TI - Well rounded. AB - Team ward rounding is a time-honored practice. Trainees learn not only clinical aspects but also ethical values of patient care. Despite current emphases on the principles of professionalism and humanism in medicine, there remains little discussion about applying these principles to ward rounds. We believe that rounds can become classrooms for the intentional inculcation of professionalism and humanism as counterweights to unintentionally promulgated brusqueness, ambivalence, cynicism, and frustration. We gathered examples of "humanistic rounding" from various institutions that should stimulate discussion and illustrate that no specialized training is required to "humanize" rounds; rather, willingness and creativity are key ingredients. PMID- 15795336 TI - Predictors and consequences of negative physician attitudes toward HIV-infected injection drug users. AB - BACKGROUND: We evaluated physicians' training, experience, and practice characteristics and examined associations between their attitudes toward human immunodeficiency virus (HIV)-infected persons who are injection drug users (IDUs) and quality of care. METHODS: Cross-sectional surveys were conducted among a probability sample of noninstitutionalized HIV-infected individuals in the United States and their main HIV care physicians. Physician and practice characteristics, training, HIV knowledge, experience, attitudes toward HIV infected IDUs, stress levels, and satisfaction with practice were assessed. The main quality-of-care measures were patient exposure to highly active antiretroviral therapy, reported problems, satisfaction with care, unmet needs, and perceived access to care. RESULTS: Nationally, 23.2% of HIV-infected patients had physicians with negative attitudes toward IDUs. Seeing more IDUs, having higher HIV treatment knowledge scores, and treating fewer patients per week were independently associated with more positive attitudes toward IDUs. Injection drug users who were cared for by physicians with negative attitudes had a significantly lower adjusted rate of exposure to highly active antiretroviral therapy by December 1996 (13.5%) than non-IDUs who were cared for by such physicians (36.1%) or IDUs who were cared for by physicians with positive attitudes (32.3%). Physician attitudes were not associated with other problems with care, satisfaction with care, unmet needs, or perceived access to care. CONCLUSIONS: Negative attitudes may lead to less than optimal care for IDUs and other marginalized populations. Providing education or experience-based exercises or ensuring that clinicians have adequate time to deal with complex problems might result in better attitudes and higher quality of care. PMID- 15795337 TI - The effect of physician disclosure of financial incentives on trust. AB - BACKGROUND: Many physicians receive financial incentives to limit their ordering of expensive tests and procedures. While Medicare mandates disclosure of incentives, it is not clear how to inform patients without undermining trust. METHODS: Our objective was to determine public opinion about physician disclosure of financial incentives and how this might be best communicated to patients. The 2002 General Social Survey included 2765 interviews from a probability sample of English-speaking US households. The interview included questions about financial incentives and an audiotaped scenario of a physician discussing the impact of financial incentives on ordering a magnetic resonance image. Respondents heard 1 of 6 randomly selected disclosure strategies. The measurements included ratings of trust, satisfaction, agreement with the physician's decision, and likelihood of remaining with the physician/health plan or seeking a second opinion. RESULTS: Nearly half (48.8%) of respondents had previously heard of financial incentives to limit test ordering. Of the respondents, 94.8% wanted to be told about incentives, at the time of enrollment in a health plan (80.5%), by a health plan representative (44.8%), their physician (17.1%), or both (38.1%). Of the 6 different disclosure strategies, "addressing emotions" and "negotiation" were associated with the best outcomes, while "common enemy" and "denying influences" were most negatively perceived. Black and Hispanic subjects were less likely to express satisfaction or trust and more likely to disenroll or seek a second opinion. CONCLUSIONS: The public wants information about physician financial incentives. Specific communication styles enhance how this information is conveyed to patients, increasing trust and supporting the physician-patient relationship. PMID- 15795338 TI - Physician strategies to reduce patients' out-of-pocket prescription costs. AB - BACKGROUND: Physicians often do not communicate with patients about out-of-pocket costs, although research indicates that physicians and patients value such discussion. METHODS: Cross-sectional national random sample mail survey of 1400 cardiologists and general internists to quantify barriers to communication about out-of-pocket costs and strategies used to assist patients in order of likelihood (from 5 [extremely likely] to 1 [not at all likely]). RESULTS: Overall, 519 (39.1%) of 1328 eligible physicians responded to the survey. The most common barriers were lack of habit, insufficient time, and concern over patient discomfort. The most common strategies used to assist patients were switching to a generic drug (mean, 4.34; SD, 0.86), using office samples (mean, 4.16; SD, 1.22), and discontinuing nonessential medicines (mean, 4.03; SD, 0.99). There were no statistically significant differences between cardiologists and general internists in barriers or strategies examined (P<.05). CONCLUSIONS: Our findings suggest that patient-physician communication about out-of-pocket costs is a problem affecting specialists and generalists nationwide. Despite barriers, physicians use multiple strategies that may vary in efficacy to assist patients burdened by these costs. PMID- 15795339 TI - Physicians' responses to resource constraints. AB - BACKGROUND: A common dilemma that confronts physicians in clinical practice is the allocation of scarce resources. Yet the strategies used by physicians in actual situations of resource constraint have not been studied. This study explores the strategies and rationales reported by physicians in situations of resource constraints encountered in practice. METHODS: A national survey of US internists, oncologists, and intensive care specialists was performed by computer assisted telephone interviews. As part of this survey, we asked physicians to tell us about a recent ethical dilemma encountered in practice. A subset of respondents reported difficulties regarding resource allocation. Transcripts of open-ended responses were coded for content based on consensus. RESULTS: Of the 600 physicians originally identified, 537 were eligible and 344 participated (response rate, 64%). Internists do not make allocation decisions alone but rather engage in negotiation in their resolution. Furthermore, these decisions are not made as dichotomous choices. Rather they often involve alternative solutions in the face of complexities of both the health care system and situations where limited resources must be allocated. Justice is not commonly the justification for rationing. CONCLUSION: Physicians' experiences in situations of resource constraints appear to be more complex than the normative literature on health care rationing assumes. In addition, reasoning about justice in health care seems to play only a small part in clinical decision making. Bridging this gap could be an important step in fostering fair allocation of resources in difficult cases. PMID- 15795340 TI - Advertising by academic medical centers. AB - BACKGROUND: Many academic medical centers have increased their use of advertising to attract patients. While the content of direct-to-consumer pharmaceutical advertisements (ads) has been studied, to our knowledge, advertising by academic medical centers has not. We aimed to characterize advertising by the nation's top academic medical centers. METHODS: We contacted all 17 medical centers named to the US News & World Report 2002 honor roll of "America's Best Hospitals" for a semistructured interview regarding their advertising practices. In addition, we obtained and systematically analyzed all non-research-related print ads placed by these institutions in their 5 most widely circulating local newspapers during 2002. RESULTS: Of the 17 institutions, 16 reported advertising to attract patients; 1 stated, "We're just word of mouth." While all 17 centers confirmed the presence of an institutional review board process for approving advertising to attract research subjects, none reported a comparable process for advertising to attract patients. We identified 127 unique non-research-related print ads for the 17 institutions during 2002 (mean, 7.5; range, 0-39). Three ads promoted community events with institution sponsorship, 2 announced genuine public services, and 122 were aimed at attracting patients. Of the latter group, 36 ads (29.5%) promoted the medical center as a whole, while 65 (53.3%) promoted specific clinical departments and 21 (17.2%) promoted single therapeutic interventions or diagnostic tests. The most commonly used marketing strategies included appealing to emotions (61.5%), highlighting institution prestige (60.7%), mentioning a symptom or disease (53.3%), and promoting introductory lectures or special offers likely to lead to further business (47.5%). Of the 21 ads for single interventions, most were for unproved (38.1%) or cosmetic (28.6%) procedures. While more than half of these ads presented benefits, none quantified their positive claims and just 1 mentioned potential harms. CONCLUSIONS: Advertising to attract patients is common among top academic medical centers but is not subjected to the oversight standard for clinical research. Many of the ads seemed to place the interests of the medical center before the interests of the patients. PMID- 15795341 TI - Research with stored biological samples: what do research participants want? AB - BACKGROUND: There is widespread disagreement about the type of consent needed for research with stored biological samples. Many believe consent for each future use is required to respect individuals. Others worry this approach may block important research. METHODS: We analyzed 1670 consent forms signed by research participants at the Warren G. Magnuson Clinical Center, National Institutes of Health, between January 1, 2000, and May 31, 2002, that offer options for future research with participants' biological samples. The research participants were healthy volunteers, family members of affected individuals, and individuals with a broad range of medical conditions enrolled in clinical research studies with and without the prospect of direct medical benefit. RESULTS: Overall, 87.1% of research participants given the option chose to authorize future research on any medical condition. More than 85% permitted unlimited future research with their stored biological samples regardless of sex, age, geographic location, or whether the individual was affected by the disease being studied or a healthy volunteer. Only 6.7% of those given the option to refuse all future research did so. Although African Americans were less likely to permit future research, 75.0% of African Americans still authorized unlimited future research with their samples. CONCLUSIONS: Most research participants authorize the unlimited future research use of their biological samples when given the opportunity to do so. These findings suggest that providing research participants with a simple binary choice to authorize or refuse all future research might allow individuals to control use of their samples, simplify consent forms, and allow important research to proceed. PMID- 15795342 TI - Frequency of physician-directed assistance for smoking cessation in patients receiving cessation medications. AB - BACKGROUND: Little is known about current physician-patient interactions regarding smoking cessation when a prescription is given for cessation medications. METHODS: We identified 1360 health plan members who were smokers and had filled prescriptions for bupropion hydrochloride or nicotine replacement products from March 26, 2002, through April 8, 2003. These members were surveyed 3 months after the order was filled to learn about the physician encounter that produced these prescriptions and the role of the physician in selecting the medication, supporting cessation, and achieving desirable outcomes. RESULTS: Among the 1035 eligible complete responses (adjusted response rate, 78%), only 36.4% reported being asked to set a specific quit date; 25.3%, to use cessation counseling; and 23.3%, to have any type of follow-up. The only physician action associated with the use of the medication was asking to set a quit date (90.7% vs 83.5%; P = .001) and no action was significantly associated with cessation. Smokers with tobacco-related chronic conditions were more likely to report physician influence but less likely to report receiving information on using the quit aid (60.6% vs 66.5%; P = .048) or being asked to set a quit date (31.7% vs 41.4%; P = .001). CONCLUSIONS: Despite the evidence-based guideline recommendations for more support for cessation, physician support actions for those receiving prescriptions appear to be relatively infrequent and to have little influence on the use of medication by smokers or on their smoking cessation. Other environmental changes may be needed to facilitate more thorough support. PMID- 15795343 TI - Improving colorectal cancer screening by targeting office systems in primary care practices: disseminating research results into clinical practice. AB - BACKGROUND: Randomized trials have shown the efficacy of an office systems approach in improving colorectal cancer (CRC) screening behaviors; its feasibility in real-world primary care practices has not been well studied. METHODS: Between August 1, 2000, and December 1, 2001, we enrolled 185 primary care clinicians identified through purchased database lists. At the end of follow up (December 31, 2002), 127 clinicians had completed preintervention and postintervention questionnaires. Trained staff from the American Cancer Society visited practices and identified areas for improvement in CRC screening. They provided clinicians with resources, tools, and support to facilitate positive change. We defined 5 clinician behavior areas related to successful CRC screening, including educating patients, identifying patients due for screening, enabling patient compliance, monitoring patient compliance, and notifying patients of their test results. We measured these areas before and after the intervention using questionnaires and data extracted from medical records. RESULTS: We demonstrated improvements in the passive use of posters and brochures about CRC screening (baseline, 20.5% and follow-up, 69.3%; P<.001) and in the monitoring of fecal occult blood tests using manual tracking systems (baseline, 20.6% and follow-up, 37.3%; P<.05). Based on medical records data among 551 patients, we found a statistically significant increase in the number of patients who became up-to-date with CRC screening recommendations and tests (P< .001 for both). CONCLUSION: Methods shown to improve CRC screening processes in protocol driven randomized trials may be effective in community practice, and wider dissemination of these strategies shows promise to increase CRC screening. PMID- 15795344 TI - Improvements in advance care planning in the Veterans Affairs System: results of a multifaceted intervention. AB - BACKGROUND: Advance care planning (ACP) aims to guide health care in the event of decisional incapacity. Interventions to promote ACP have had limited effectiveness. We conducted an educational and motivational intervention in Department of Veterans Affairs outpatient clinics to increase ACP use and proxy and health care provider understanding of patients' preferences and values. METHODS: We recruited 23 providers and up to 14 of each of their patients; the patients were randomized to the control or intervention group. Eligibility criteria included a preexisting relationship with the provider, age 55 years or older, chronic health condition(s), and no recorded advance directive. The intervention group (n = 119) received an ACP workbook, motivational counseling by social workers, and cues to providers to discuss ACP. The control group (n = 129) received an advance directive booklet. RESULTS: The intervention patients reported more ACP discussions with their providers (64% vs 38%; P<.001). Living wills were filed in the medical record twice as often in the intervention group (48% vs 23%; P<.001). Provider-patient dyads in the intervention group had higher agreement scores than the control group for treatment preferences, values, and personal beliefs (58% vs 48%, 57% vs 46%, and 61% vs 47%, respectively; P<.01 for all comparisons). The agreement scores for the proxy-patient dyads did not differ between groups for treatment preferences and values, but were higher in the intervention than the control group for personal beliefs (67% vs 56%). CONCLUSION: This intervention demonstrates mixed results and highlights the ongoing challenges of helping health care providers and potential proxy decision makers represent patient preferences and values. PMID- 15795345 TI - New Alstrom syndrome phenotypes based on the evaluation of 182 cases. AB - BACKGROUND: Alstrom syndrome is a recessively inherited genetic disorder characterized by congenital retinal dystrophy that leads to blindness, hearing impairment, childhood obesity, insulin resistance, and type 2 diabetes mellitus. We provide new details on cardiologic, hepatic, gastrointestinal, urologic, pulmonary, and neurobehavioral phenotypes in Alstrom syndrome and describe the histopathologic findings in 5 individuals. METHODS: We obtained data on 182 patients from clinical examinations, medical record reviews, standardized questionnaires, and personal interviews with physicians and parents. RESULTS: Dilated cardiomyopathy occurred in 60% of patients. Age at onset was either during infancy, often before vision disturbances were noted, or in adolescence or adulthood. There is a risk of recurrence of infantile cardiomyopathy. Hyperinsulinemia (92%) developed in early childhood and progressed to type 2 diabetes mellitus in 82% of those older than 16 years. Hypertriglyceridemia (54%) precipitated pancreatitis in 8 patients. Urologic dysfunction and gastrointestinal disturbances occurred in 48% and 35% of patients, respectively. Fifty-three percent of patients had persistent pulmonary symptoms. Neurologic symptoms in 20% of patients included clonic tic and absence seizures. Developmental motor or language delays were observed in 46% of patients. Fibrotic infiltrations of multiple organs, that is, kidney, heart, liver, lung, urinary bladder, gonads, and pancreas, were observed. CONCLUSIONS: The wide-ranging and complex spectrum of phenotypes reported herein broadens those previously described for Alstrom syndrome. These findings will aid physicians in making an early and accurate diagnosis and will help effect appropriate monitoring and treatment. PMID- 15795346 TI - Low bone mass in subjects on a long-term raw vegetarian diet. AB - BACKGROUND: Little is known regarding the health effects of a raw food (RF) vegetarian diet. METHODS: We performed a cross-sectional study on 18 volunteers (mean +/- SD age, 54.2 +/- 11.5 years; male/female ratio, 11:7) on a RF vegetarian diet for a mean of 3.6 years and a comparison age- and sex-matched group eating typical American diets. We measured body composition, bone mineral content and density, bone turnover markers (C-telopeptide of type I collagen and bone-specific alkaline phosphatase), C-reactive protein, 25-hydroxyvitamin D, insulin-like growth factor 1, and leptin in serum. RESULTS: The RF vegetarians had a mean +/- SD body mass index (calculated as weight in kilograms divided by the square of height in meters) of 20.5 +/- 2.3, compared with 25.4 +/- 3.3 in the control subjects. The mean bone mineral content and density of the lumbar spine (P= .003 and P<.001, respectively) and hip (P = .01 and P<.001, respectively) were lower in the RF group than in the control group. Serum C telopeptide of type I collagen and bone-specific alkaline phosphatase levels were similar between the groups, while the mean 25-hydroxyvitamin D concentration was higher in the RF group than in the control group (P<.001). The mean serum C reactive protein (P = .03), insulin-like growth factor 1 (P = .002), and leptin (P = .005) were lower in the RF group. CONCLUSION: A RF vegetarian diet is associated with low bone mass at clinically important skeletal regions but is without evidence of increased bone turnover or impaired vitamin D status. PMID- 15795348 TI - Human immunodeficiency virus 1 infection, cocaine, and coronary calcification. AB - BACKGROUND: Although cocaine use and human immunodeficiency virus (HIV) infection have been linked with clinical cardiovascular disease, the effects of cocaine use and HIV infection, especially the combination of the 2, on subclinical disease have rarely been reported. The objective of this study was to evaluate whether cocaine use alone, HIV infection alone, or a combination of the 2 is associated with coronary calcification, a marker of subclinical atherosclerosis. METHODS: Between May 20, 2000, and March 31, 2003, 224 black study participants from Baltimore were enrolled in an observational study of subclinical atherosclerosis as related to HIV and cocaine use. Interviews about sociodemographic characteristics and drug use behaviors, clinical examinations, echocardiographic examinations, lipid profiles, high-sensitivity C-reactive protein tests, and computed tomographic scans for coronary calcium were performed. Although the overall investigation is a cohort study, the data presented herein are cross sectional only. RESULTS: The highest proportion (37.6%) of presence of coronary calcification was in the HIV-positive and cocaine-positive group, followed by 29.8% in the HIV-negative and cocaine-positive group, 28.6% in the HIV-positive and cocaine-negative group, and 18.8% in the HIV-negative and cocaine-negative group. Univariate analysis showed that HIV, cocaine use, and both were associated with a higher number of lesions, calcified area, volume, and calcium score. In multiple regression analysis with adjustment for age, body mass index, low density lipoprotein cholesterol level, triglyceride level, mean corpuscular volume, and systolic blood pressure, HIV, cocaine use, and both were independently associated with coronary calcification. CONCLUSION: These results suggest that HIV infection alone, cocaine use alone, or the 2 combined may contribute to early subclinical atherosclerotic cardiovascular disease. PMID- 15795350 TI - Unreimbursed services in primary care. PMID- 15795349 TI - Conventional and atypical antipsychotics and the risk of hospitalization for ventricular arrhythmias or cardiac arrest. AB - BACKGROUND: Conventional antipsychotic drugs have been implicated as a cause of ventricular arrhythmias and cardiac arrest, but no definitive information is available regarding atypical antipsychotics. We compared the effect of conventional and atypical antipsychotics on the risk of hospitalization for ventricular arrhythmias and cardiac arrest. METHODS: We conducted a case-control study on residents of nursing homes in 6 US states by using data from the Minimum Data Set linked to Medicare inpatient claims. Cases were residents hospitalized for ventricular arrhythmias or cardiac arrest between July 4, 1998, and December 30, 1999. For each case, we identified up to 5 controls residing in the same facility during the same period. The sample consisted of 649 cases and 2962 controls. RESULTS: Use of conventional antipsychotics was associated with a nearly 2-fold increase in risk of hospitalization for ventricular arrhythmias or cardiac arrest (adjusted odds ratio, 1.86; 95% confidence interval, 1.27-2.74). There was no increased risk associated with the use of atypical antipsychotics (odds ratio, 0.87; 95% confidence interval, 0.58-1.32). The risk of hospitalization for ventricular arrhythmias or cardiac arrest was highest among conventional users with cardiac disease (odds ratio, 3.27; 95% confidence interval, 1.95-5.47). However, cardiac disease and conventional antipsychotics did not show a synergistic effect (synergy index, 1.19). CONCLUSIONS: Conventional but not atypical antipsychotics are associated with an increased risk of hospitalization for ventricular arrhythmias and cardiac arrest. The prescription of conventional antipsychotics in patients with cardiac disease should be carefully weighed. PMID- 15795351 TI - The likelihood that PSA detection of early prostate cancers is associated with high mortality. PMID- 15795352 TI - Capsaicin cream unpopular with patients. PMID- 15795353 TI - The MEDLINE search as a diagnostic maneuver. PMID- 15795354 TI - Factors associated with INR elevation and bleeding complications during Warfarin therapy. PMID- 15795355 TI - Diagnostic accuracy of natriuretic peptides for heart failure. PMID- 15795356 TI - Images in cardiovascular medicine. A "mickey mouse" coronary anomaly: aorto-left atrial fistula with aneurysm. PMID- 15795357 TI - Letter regarding articles by Kereiakes and Willerson, Mehran et al, and Kaplan et al, "mini-review: expert opinion". PMID- 15795358 TI - Letter regarding article by Khan et al, "predictive adaptive responses to maternal high-fat diet prevent endothelial dysfunction but not hypertension in adult rat offspring". PMID- 15795359 TI - Letter regarding article by Vita et al, "serum myeloperoxidase levels independently predict endothelial dysfunction in humans". PMID- 15795360 TI - Can hysterectomy be considered a risk factor for cardiovascular disease? PMID- 15795361 TI - Nesiritide and worsening of renal function: the emperor's new clothes? PMID- 15795362 TI - Carbon dioxide-rich water bathing enhances collateral blood flow in ischemic hindlimb via mobilization of endothelial progenitor cells and activation of NO cGMP system. AB - BACKGROUND: Carbon dioxide-rich water bathing has the effect of vasodilatation, whereas it remains undetermined whether this therapy exerts an angiogenic action associated with new vessel formation. METHODS AND RESULTS: Unilateral hindlimb ischemia was induced by resecting the femoral arteries of C57BL/J mice. Lower limbs were immersed in CO2-enriched water (CO2 concentration, 1000 to 1200 mg/L) or freshwater (control) at 37 degrees C for 10 minutes once a day. Laser Doppler imaging revealed increased blood perfusion in ischemic limbs of CO2 bathing (38% increase at day 28, P<0.001), whereas N(G)-nitro-L-arginine methyl ester treatment abolished this effect. Angiography or immunohistochemistry revealed that collateral vessel formation and capillary densities were increased (4.1-fold and 3.7-fold, P<0.001, respectively). Plasma vascular endothelial growth factor (VEGF) levels were elevated at day 14 (18%, P<0.05). VEGF mRNA levels, phosphorylation of NO synthase, and cGMP accumulation in the CO2-bathed hindlimb muscles were increased (2.7-fold, 2.4-fold, and 3.4-fold, respectively) but not in forelimb muscles. The number of circulating Lin-/Flk-1+/CD34- endothelial lineage progenitor cells was markedly increased by CO2 bathing (24-fold at day 14, P<0.001). The Lin-/Flk-1+/CD34- cells express other endothelial antigens (endoglin and VE-cadherin) and incorporated acetylated LDL. CONCLUSIONS: Our present study demonstrates that CO2 bathing of ischemic hindlimb causes the induction of local VEGF synthesis, resulting in an NO-dependent neocapillary formation associated with mobilization of endothelial progenitor cells. PMID- 15795364 TI - Angiogenesis: where do we stand now? PMID- 15795363 TI - Inflammation and endothelial function: direct vascular effects of human C reactive protein on nitric oxide bioavailability. AB - BACKGROUND: Circulating concentrations of the sensitive inflammatory marker C reactive protein (CRP) predict future cardiovascular events, and CRP is elevated during sepsis and inflammation, when vascular reactivity may be modulated. We therefore investigated the direct effect of CRP on vascular reactivity. METHODS AND RESULTS: The effects of isolated, pure human CRP on vasoreactivity and protein expression were studied in vascular rings and cells in vitro, and effects on blood pressure were studied in rats in vivo. The temporal relationship between changes in CRP concentration and brachial flow-mediated dilation was also studied in humans after vaccination with Salmonella typhi capsular polysaccharide, a model of inflammatory endothelial dysfunction. In contrast to some previous reports, highly purified and well-characterized human CRP specifically induced hyporeactivity to phenylephrine in rings of human internal mammary artery and rat aorta that was mediated through physiological antagonism by nitric oxide (NO). CRP did not alter endothelial NO synthase protein expression but increased protein expression of GTP cyclohydrolase-1, the rate-limiting enzyme in the synthesis of tetrahydrobiopterin, the NO synthase cofactor. In the vaccine model of inflammatory endothelial dysfunction in humans, increased CRP concentration coincided with the resolution rather than the development of endothelial dysfunction, consistent with the vitro findings; however, administration of human CRP to rats had no effect on blood pressure. CONCLUSIONS: Pure human CRP has specific, direct effects on vascular function in vitro via increased NO production; however, further clarification of the effect, if any, of CRP on vascular reactivity in humans in vivo will require clinical studies using specific inhibitors of CRP. PMID- 15795366 TI - How ion channels sense membrane potential. PMID- 15795365 TI - Effects of peripheral chemoreceptors deactivation on sympathetic activity in heart transplant recipients. AB - Heart transplantation initially normalizes sympathetic hyperactivity directed at the muscle circulation. However, sympathetic activity increases with time after transplantation and the exact mechanisms responsible for sympathetic control in heart transplant recipients remain unclear. We examined the effects of peripheral chemoreflex deactivation caused by breathing 100% oxygen on muscle sympathetic nerve activity (expressed as number of burst per minute and mean burst amplitude), heart rate, and mean blood pressure in 13 heart transplant recipients, 13 patients with essential hypertension, and 10 controls. Heart transplant recipients disclosed the highest sympathetic activity, whereas it did not differ between controls and patients with essential hypertension (51+/-16 versus 37+/-14 versus 39+/-12 burst/min, respectively; P<0.05). Breathing 100% oxygen, in comparison with 21% oxygen, reduced sympathetic activity (-4+/-4 versus -1+/-2 burst/min, P<0.01; 85+/-9 versus 101+/-8% of amplitude at baseline, P<0.001) and mean blood pressure (-4+/-5 versus +3+/-6 mm Hg; P<0.05) in heart transplant recipients, decreased sympathetic activity (-4+/-4 versus 0+/-3 burst/min, P<0.05; 90+/-16 versus 101+/-9% of amplitude at baseline, P<0.05) in patients with essential hypertension, but did not reduce sympathetic activity (2+/-4 versus 3+/-3 burst/min, P=NS; 95+/-11 versus 95+/-13% of amplitude at baseline, P=NS) in control subjects. The sympathetic response to hyperoxia was more marked in heart transplant recipients than in controls (85+/-9 versus 95+/ 11% of baseline amplitude; P<0.05). The decrease in sympathetic activity was most evident in patients with the longest time after heart transplantation (r=-0.75, P<0.01). In conclusion, tonic chemoreflex activation increases resting muscle sympathetic nerve activity and favors blood pressure elevation after heart transplantation. PMID- 15795367 TI - Sigma cascades in prokaryotic regulatory networks. PMID- 15795368 TI - A simple answer for a splicing conundrum. PMID- 15795369 TI - Thrips and tospoviruses come of age: mapping determinants of insect transmission. PMID- 15795370 TI - Targeting of herpesvirus capsid transport in axons is coupled to association with specific sets of tegument proteins. AB - The capsids of neurotropic herpesviruses have the remarkable ability to move in specific directions within axons. By modulating bidirectional capsid transport to favor either retrograde (minus-end) or anterograde (plus-end) motion, these viruses travel to sensory ganglia or peripheral tissue at specific stages of infection. By using correlative motion analysis to simultaneously monitor the trafficking of distinct viral proteins in living neurons, we demonstrate that viral "tegument" proteins are complexed to capsids moving in axons. The removal of a subset of tegument proteins from capsids invariably preceded retrograde transport to the cell body in sensory ganglia, whereas addition of these proteins was coupled to anterograde transport of progeny capsids to the distal axon. Although capsid transport never occurred without associated tegument proteins, anterograde-specific tegument proteins were competent to travel to the distal axon independent of capsids. These findings are compatible with a model of viral bidirectional transport in which tegument proteins direct capsid traffic to specific intracellular locations during the infectious cycle. PMID- 15795371 TI - Genomic characterization reveals a simple histone H4 acetylation code. AB - The histone code hypothesis holds that covalent posttranslational modifications of histone tails are interpreted by the cell to yield a rich combinatorial transcriptional output. This hypothesis has been the subject of active debate in the literature. Here, we investigated the combinatorial complexity of the acetylation code at the four lysine residues of the histone H4 tail in budding yeast. We constructed yeast strains carrying all 15 possible combinations of mutations among lysines 5, 8, 12, and 16 to arginine in the histone H4 tail, mimicking positively charged, unacetylated lysine states, and characterized the resulting genome-wide changes in gene expression by using DNA microarrays. Only the lysine 16 mutation had specific transcriptional consequences independent of the mutational state of the other lysines (affecting approximately 100 genes). In contrast, for lysines 5, 8, and 12, expression changes were due to nonspecific, cumulative effects seen as increased transcription correlating with an increase in the total number of mutations (affecting approximately 1,200 genes). Thus, acetylation of histone H4 is interpreted by two mechanisms: a specific mechanism for lysine 16 and a nonspecific, cumulative mechanism for lysines 5, 8, and 12. PMID- 15795372 TI - Quantitative analysis of binding motifs mediating diverse spatial readouts of the Dorsal gradient in the Drosophila embryo. AB - Dorsal is a sequence-specific transcription factor that is distributed in a broad nuclear gradient across the dorsal-ventral (DV) axis of the early Drosophila embryo. It initiates gastrulation by regulating at least 30-50 target genes in a concentration-dependent fashion. Previous studies identified 18 enhancers that are directly regulated by different concentrations of Dorsal. Here, we employ computational methods to determine the basis for these distinct transcriptional outputs. Orthologous enhancers were identified in a variety of divergent Drosophila species, and their comparison revealed several conserved sequence features responsible for DV patterning. In particular, the quality of Dorsal and Twist recognition sequences correlates with the DV coordinates of gene expression relative to the Dorsal gradient. These findings are entirely consistent with a gradient threshold model for DV patterning, whereby the quality of individual Dorsal binding sites determines in vivo occupancy of target enhancers by the Dorsal gradient. Linked Dorsal and Twist binding sites constitute a conserved composite element in certain "type 2" Dorsal target enhancers, which direct gene expression in ventral regions of the neurogenic ectoderm in response to intermediate levels of the Dorsal gradient. Similar motif arrangements were identified in orthologous loci in the distant mosquito genome, Anopheles gambiae. We discuss how Dorsal and Twist work either additively or synergistically to activate different target enhancers. PMID- 15795373 TI - Identifying Foxp3-expressing suppressor T cells with a bicistronic reporter. AB - Regulatory T cells are critical for maintaining self-tolerance and to negatively regulate immune responses. Foxp3 is a regulatory T cell-specific transcription factor that functions as the master regulator of the development and function of regulatory T cells. Here, we report the generation of a mouse model, in which a bicistronic reporter expressing a red fluorescent protein has been knocked into the endogenous Foxp3 locus. Using this mouse model, we assessed Foxp3 expression in various lymphocyte compartments and identified previously unreported Foxp3 expressing cells. In addition, we showed that de novo Foxp3 expression along with suppressive function were induced by TGF-beta in activated CD4 T cells in vitro. Finally, we demonstrated that non-Foxp3-expressing CD4 T cells could not be converted into Foxp3-expressing cells upon adoptive transfer into immunodeficient hosts. This Foxp3 bicistronic reporter knockin mouse model should greatly enhance the study of regulation and function of Foxp3-expressing regulatory T cells. PMID- 15795374 TI - A unique loop in T7 DNA polymerase mediates the binding of helicase-primase, DNA binding protein, and processivity factor. AB - Bacteriophage T7 DNA polymerase (gene 5 protein, gp5) interacts with its processivity factor, Escherichia coli thioredoxin, via a unique loop at the tip of the thumb subdomain. We find that this thioredoxin-binding domain is also the site of interaction of the phage-encoded helicase/primase (gp4) and ssDNA binding protein (gp2.5). Thioredoxin itself interacts only weakly with gp4 and gp2.5 but drastically enhances their binding to gp5. The acidic C termini of gp4 and gp2.5 are critical for this interaction in the absence of DNA. However, the C-terminal tail of gp4 is not required for binding to gp5 when the latter is bound to a primer/template. We propose that the thioredoxin-binding domain is a molecular switch that regulates the interaction of T7 DNA polymerase with other proteins of the replisome. PMID- 15795376 TI - Generalized Bienenstock-Cooper-Munro rule for spiking neurons that maximizes information transmission. AB - Maximization of information transmission by a spiking-neuron model predicts changes of synaptic connections that depend on timing of pre- and postsynaptic spikes and on the postsynaptic membrane potential. Under the assumption of Poisson firing statistics, the synaptic update rule exhibits all of the features of the Bienenstock-Cooper-Munro rule, in particular, regimes of synaptic potentiation and depression separated by a sliding threshold. Moreover, the learning rule is also applicable to the more realistic case of neuron models with refractoriness, and is sensitive to correlations between input spikes, even in the absence of presynaptic rate modulation. The learning rule is found by maximizing the mutual information between presynaptic and postsynaptic spike trains under the constraint that the postsynaptic firing rate stays close to some target firing rate. An interpretation of the synaptic update rule in terms of homeostatic synaptic processes and spike-timing-dependent plasticity is discussed. PMID- 15795375 TI - 23S rRNA base pair 2057-2611 determines ketolide susceptibility and fitness cost of the macrolide resistance mutation 2058A-->G. AB - The 23S rRNA A2058G alteration mediates macrolide, lincosamide, and streptogramin B resistance in the bacterial domain and determines the selectivity of macrolide antibiotics for eubacterial ribosomes, as opposed to eukaryotic ribosomes. However, this mutation is associated with a disparate resistance phenotype: It confers high-level resistance to ketolides in mycobacteria but only marginally affects ketolide susceptibility in streptococci. We used site-directed mutagenesis of nucleotides within domain V of 23S rRNA to study the molecular basis for this disparity. We show that mutational alteration of the polymorphic 2057-2611 base pair from A-U to G-C in isogenic mutants of Mycobacterium smegmatis significantly affects susceptibility to ketolides but does not influence susceptibility to other macrolide antibiotics. In addition, we provide evidence that the 2057-2611 polymorphism determines the fitness cost of the 23S rRNA A2058G resistance mutation. Supported by structural analysis, our results indicate that polymorphic nucleotides mediate the disparate phenotype of genotypically identical resistance mutations and provide an explanation for the large species differences in the epidemiology of defined drug resistance mutations. PMID- 15795377 TI - Perceptual learning in clear displays optimizes perceptual expertise: learning the limiting process. AB - Human operators develop expertise in perceptual tasks by practice or perceptual learning. For noisy displays, practice improves performance by learned external noise filtering. For clear displays, practice improves performance by improved amplification or enhancement of the stimulus. Can these two mechanisms of perceptual improvement be trained separately? In an orientation task, we found that training with clear displays generalized to performance in noisy displays, but we did not find the reverse to be true. In noisy displays, the noise in the stimulus limits performance. In clear displays, performance is limited by noisiness of internal representations and processes. Our results suggest that training in one display condition optimizes the limiting factor(s) in performance in that condition and that noise filtering is also improved by exposure to the stimulus in clear displays. The asymmetric pattern of transfer implies the existence of two independent mechanisms of perceptual learning, which may reflect channel re-weighting in adult visual system. These results also suggest that training operators with clear stimuli may suffice to improve performance in a range of clear and noisy environments by simultaneous learning by two mechanisms. PMID- 15795378 TI - Xenopus as a model system to study transcriptional regulatory networks. AB - Development is controlled by a complex series of events requiring sequential gene activation. Understanding the logic of gene networks during development is necessary for a complete understanding of how genes contribute to phenotype. Pioneering work initiated in the sea urchin and Drosophila has demonstrated that reasonable transcriptional regulatory network diagrams representing early development in multicellular animals can be generated through use of appropriate genomic, genetic, and biochemical tools. Establishment of similar regulatory network diagrams for vertebrate development is a necessary step. The amphibian Xenopus has long been used as a model for vertebrate early development and has contributed greatly to the elucidation of gene regulation. Because the best and most extensively studied transcriptional regulatory network in Xenopus is that underlying the formation and function of Spemann's organizer, we describe the current status of our understanding of this gene regulatory network and its relationship to mesodermal patterning. Seventy-four transcription factors currently known to be expressed in the mesoendoderm of Xenopus gastrula were characterized according to their modes of action, DNA binding consensus sequences, and target genes. Among them, nineteen transcription factors were characterized sufficiently in detail, allowing us to generate a gene regulatory network diagram. Additionally, we discuss recent amphibian work using a combined DNA microarray and bioinformatics approach that promises to accelerate regulatory network studies. PMID- 15795380 TI - High-resolution functional proteomics by active-site peptide profiling. AB - Characterization and functional annotation of the large number of proteins predicted from genome sequencing projects poses a major scientific challenge. Whereas several proteomics techniques have been developed to quantify the abundance of proteins, these methods provide little information regarding protein function. Here, we present a gel-free platform that permits ultrasensitive, quantitative, and high-resolution analyses of protein activities in proteomes, including highly problematic samples such as undiluted plasma. We demonstrate the value of this platform for the discovery of both disease-related enzyme activities and specific inhibitors that target these proteins. PMID- 15795379 TI - Intracellular Helicobacter pylori in gastric epithelial progenitors. AB - Helicobacter pylori is generally viewed as an extracellular pathogen. We have analyzed the tropism of H. pylori clinical isolates in a gnotobiotic transgenic mouse model of human chronic atrophic gastritis, a preneoplastic condition. These mice lack acid-producing parietal cells and have an amplified population of dividing gastric epithelial progenitors (GEPs) that express NeuAc alpha 2,3Gal beta 1,4-glycans recognized by H. pylori adhesins. Scanning confocal and transmission electron microscopic studies of stomachs that had been colonized for 1 month or 1 year revealed intracellular bacterial collections (IBCs) in a small subset of multi- and oligopotential epithelial progenitors. Transmission electron microscopic and multilabel immunohistochemical analyses disclosed bacteria with several morphotypes, including spiral-shaped, in the cytoplasm and endosomes. Several stages in IBC evolution were documented, from a few solitary bacteria to consolidated populations in dividing and nondividing GEPs, to microorganisms traversing breaches in the GEP plasma cell membrane. IBC formation was not a unique feature of H. pylori strains isolated from patients with chronic atrophic gastritis. The notion that adult mammalian epithelial progenitors can function as a repository for H. pylori broadens the view of host habitats available to this and perhaps other pathogens. PMID- 15795381 TI - Relationship between follicle size at insemination and pregnancy success. AB - Administration of gonadotropin-releasing hormone (GnRH) induces a surge of luteinizing hormone and ovulation in a variety of species, including human beings. Our objectives were to determine the effect of follicle size at the time of ovulation on corpus luteum function and establishment and maintenance of pregnancy in cows in which ovulation was either spontaneous or induced with GnRH. GnRH-induced ovulation of follicles < or approximately = 11 mm in diameter resulted in decreased pregnancy rates and increased late embryonic mortality. This decrease in fertility was associated with lower circulating concentrations of estradiol on the day of insemination, a decreased rate of increase in progesterone after insemination, and, ultimately, decreased circulating concentrations of progesterone. In contrast, ovulatory follicle size had no apparent effect on fertility when ovulation occurred spontaneously. Follicles undergoing spontaneous ovulation do so at a wide range of sizes when they are physiologically mature. Therefore, administration of GnRH to induce ovulation likely initiates a preovulatory gonadotropin surge before some dominant follicles attain physiological maturity. GnRH-induced ovulation of follicles that are physiologically immature has a negative impact on pregnancy rates and late embryonic/fetal survival. These observations in cattle may have implications for assisted reproductive procedures in human beings. PMID- 15795382 TI - Visual working memory in decision making by honey bees. AB - The robustness and plasticity of working memory were investigated in honey bees by using a delayed matching-to-sample (DMTS) paradigm. The findings are summarized as follows: first, performance in the DMTS task decreases as the duration between the presentation of the sample stimulus and the presentation of the comparison stimuli is increased. This decrease is well approximated by an exponential decay function. Performance is significantly better than random choice level even at delays as long as 5 sec and is reduced to random-choice levels at an average delay time of 8.68 +/- 0.06 sec. Second, when the DMTS task involves two samples (one relevant, the other irrelevant), bees can be trained to learn to use the relevant sample to perform the task if (i) the relevant sample is always at a fixed position, or (ii) the relevant sample always has the same place in the sequence of presentation (always first or always second). Bees that have learned to use the relevant sample and to ignore the irrelevant sample can generalize this learning, and apply it to novel sets of sample and comparison stimuli that they have never previously encountered. The findings point to a remarkably robust, and yet plastic, working memory in the honey bee. PMID- 15795383 TI - Defining the role of active-site loop fluctuations in dihydrofolate reductase catalysis. AB - Dynamic processes are implicit in the catalytic function of all enzymes. To obtain insights into the relationship between the dynamics and thermodynamics of protein fluctuations and catalysis, we have measured millisecond time scale motions in the enzyme dihydrofolate reductase using NMR relaxation methods. Studies of a ternary complex formed from the substrate analog folate and oxidized NADP+ cofactor revealed conformational exchange between a ground state, in which the active site loops adopt a closed conformation, and a weakly populated (4.2% at 30 degrees C) excited state with the loops in the occluded conformation. Fluctuations between these states, which involve motions of the nicotinamide ring of the cofactor into and out of the active site, occur on a time scale that is directly relevant to the structural transitions involved in progression through the catalytic cycle. PMID- 15795384 TI - Discovery of the ergothioneine transporter. AB - Variants of the SLC22A4 gene are associated with susceptibility to rheumatoid arthritis and Crohn's disease. SLC22A4 codes for an integral membrane protein, OCTN1, that has been presumed to carry organic cations like tetraethylammonium across the plasma membrane. Here, we show that the key substrate of this transporter is in fact ergothioneine (ET). Human OCTN1 was expressed in 293 cells. A substrate lead, stachydrine (alias proline betaine), was identified by liquid chromatography MS difference shading, a new substrate search strategy. Analysis of transport efficiency of stachydrine-related solutes, affinity, and Na+ dependence indicates that the physiological substrate is ET. Efficiency of transport of ET was as high as 195 microl per min per mg of protein. By contrast, the carnitine transporter OCTN2 from rat did not transport ET at all. Because ET is transported >100 times more efficiently than tetraethylammonium and carnitine, we propose the functional name ETT (ET transporter) instead of OCTN1. ET, all of which is absorbed from food, is an intracellular antioxidant with metal ion affinity. Its particular purpose is unresolved. Cells with expression of ETT accumulate ET to high levels and avidly retain it. By contrast, cells lacking ETT do not accumulate ET, because their plasma membrane is virtually impermeable for this compound. The real-time PCR expression profile of human ETT, with strong expression in CD71+ cells, is consistent with a pivotal function of ET in erythrocytes. Moreover, prominent expression of ETT in monocytes and SLC22A4 polymorphism associations suggest a protective role of ET in chronic inflammatory disorders. PMID- 15795387 TI - Emergency contraception moves behind the counter. PMID- 15795385 TI - Hydrogen peroxide generated extracellularly by receptor-ligand interaction facilitates cell signaling. AB - Reactive oxygen species (ROS) are key components of postreceptor intracellular signaling pathways; however, the role of ROS in signal initiation is uncertain. We discovered that receptor-ligand interaction caused the generation of hydrogen peroxide (H2O2). Using members of the hematopoietin receptor superfamily, as well as EGF receptor, we show that H2O2 is generated by specific receptor-ligand interaction in cells and in cell-free systems. With cognate ligand, the extracellular domain of the receptor was sufficient for H2O2 generation. We also found that production of H2O2 was diminished in a granulocyte-macrophage colony stimulating factor receptor mutant unable to bind ligand. Exogenously added H2O2 induced signaling in the absence of ligand, whereas catalase and a membrane-bound peroxiredoxin inhibited ligand-dependent signaling. Our results suggest that H2O2 produced by receptor-ligand interaction is involved as a chemical mediator that facilitates cell signaling. PMID- 15795386 TI - Cytolysin-dependent evasion of lysosomal killing. AB - Local host defenses limit proliferation and systemic spread of pathogenic bacteria from sites of mucosal colonization. For pathogens such as streptococci that fail to grow intracellularly, internalization and killing by epithelial cells contribute to the control of bacterial growth and dissemination. Here, we show that group A Streptococcus (GAS), the agent of streptococcal sore throat and invasive soft tissue infections, evades internalization and intracellular killing by pharyngeal epithelial cells. Production of the cholesterol-binding cytotoxin streptolysin O (SLO) prevented internalization of GAS into lysosomes. In striking contrast, GAS rendered defective in production of SLO were internalized directly or rapidly transported into lysosomes, where they were killed by a pH-dependent mechanism. Because SLO is the prototype of cholesterol-dependent cytolysins produced by many Gram-positive bacteria, cytolysin-mediated evasion of lysosomal killing may be a general mechanism to protect such pathogens from clearance by host epithelial cells. PMID- 15795389 TI - Perils of systematic reviews. PMID- 15795390 TI - Infant mortality in Alberta and all of Canada. PMID- 15795391 TI - Medical education and chronic disease. PMID- 15795393 TI - Outcome reporting bias in government-funded RCTs. PMID- 15795399 TI - Nonprescription status for emergency contraception. PMID- 15795401 TI - Minority government commits 20.2 billion dollars to health. PMID- 15795402 TI - Canadian Society for Vascular Surgery consensus statement on endovascular aneurysm repair. PMID- 15795403 TI - Young blood heals old muscles. PMID- 15795404 TI - "Avoid the crush": hazards of medication administration in patients with dysphagia or a feeding tube. PMID- 15795405 TI - Dioxin contamination and poisoning. PMID- 15795406 TI - A 60-year-old man with chest pain following pacemaker implantation. PMID- 15795407 TI - Does isolation prevent the spread of methicillin-resistant Staphylococcus aureus? PMID- 15795409 TI - Travel patterns and risk behaviour of HIV-positive people travelling internationally. AB - BACKGROUND: International travel is associated with an increased risk of enteric, vector-borne, sexually transmitted and blood-borne infections. These risks are even higher among immunocompromised people, such as those with HIV infection. We conducted a study to determine HIV-positive people's travel patterns and risk behaviours while abroad. METHODS: We conducted an anonymous survey of HIV positive people attending an HIV clinic in a tertiary care hospital in Toronto about their travel activities and pretravel precautions as well as their burden of illness and risk exposure during travel. We compared the characteristics of respondents who had travelled outside Canada and the United States (international travellers) with those of respondents who had not travelled internationally. RESULTS: Of 290 HIV-positive people who participated in the study, 133 (45.9%) indicated that they had travelled internationally in the 5 years before the survey. These people were predominantly men (93.2%) and well educated (60.0% had a university level education), and they had travelled mostly for personal reasons (89.5%) on trips that lasted 3.6 weeks on average. Only 58 (43.6%) sought health advice before travelling, and only 17 (12.8%) sought advice from a travel clinic. Five (3.8%) had received live vaccines before travel, and 9 (6.8%) had taken malaria chemoprophylaxis. Of the 119 international travellers who were taking antiretroviral therapy; 35 (29.4%) reported either discontinuing their medications or being poorly compliant with the therapy while travelling. Thirty one (23.3%) of the 133 international travellers reported having had casual sexual activity with new partners while travelling, and only 18 (58.1%) of them reported always using a condom. Twenty-one (15.8%) of the 133 reported having had risky exposure to sharps. Twenty-four (18.0%) said they had become ill enough while travelling to require medical attention. INTERPRETATION: Only one-fifth of HIV positive people surveyed who travelled internationally sought advice from a health professional before their trip. Their travel was associated with poor adherence to antiretroviral therapy, risky sexual practices and risky exposure to sharps. PMID- 15795408 TI - Effects of making emergency contraception available without a physician's prescription: a population-based study. AB - BACKGROUND: Timely access to emergency contraception has the potential to reduce the number of unwanted pregnancies and subsequent abortions. A public health policy initiative in British Columbia beginning in December 2000 allowed pharmacists to provide emergency contraceptives (ECs) without a prescription. We sought to determine changes in EC use after the policy was introduced and to analyze EC use with data generated by the policy. METHODS: All Ovral, Preven and Plan B EC prescriptions from Jan. 1, 1996, to Dec. 31, 2003, were identified through the BC PharmaNet and Medical Services Plan administrative databases and the data analyzed to determine changes between 1996 and 2002. Changes over time were determined in the frequency of EC provision, choice of EC agent, frequency of EC use by age group, repeat use and geographic distribution of EC prescription for the pre- and post-policy periods. Anonymized patient-specific data from treatment consent forms were used to describe the reason for EC use, interval between unprotected intercourse and EC prescription, proportion prescribed for immediate or future use, referrals for regular birth control and STD screening and concomitant antiemetic use. Consent data also provided the time in the menstrual cycle that the EC was requested. RESULTS: The number of EC prescriptions increased from a pre-policy mean of 8805 (99% confidence interval 7823-9787) in the years 1996 to 2000 to a post-policy total in 2002 of 17 794. Physicians prescribed the levonorgestrel regimen (Plan B) less frequently than did pharmacists. The frequency of EC use was highest among women aged 20-24 years across all study years, and all age groups demonstrated a post-policy increase in use. On average, 2.1% of the women received an EC 3 or more times a year over the period of the study. More women in urban regions received ECs than women in more rural areas of the province. Analysis of pharmacist treatment consent forms used in 2001 and 2002 showed that 56.2% of women receiving an EC reported using a method of birth control that had failed, 55.7% of pharmacist-provided ECs were obtained within 24 hours after unprotected intercourse, 1.1% of ECs were obtained for future use, antiemetics were provided to 57.7% of women receiving the Yuzpe regimen (Ovral, Preven) and to 20.5% of women receiving levonorgestrel, and women tended to seek ECs when unprotected intercourse occurred at the time of highest risk of pregnancy in their menstrual cycle. Women in greatest financial need obtained ECs more frequently from physicians than from pharmacists. INTERPRETATION: The policy change that granted pharmacists authority to provide ECs to women without a physician's prescription did not simply expand EC availability but was associated with an overall increase in EC use in the province. PMID- 15795410 TI - Supporting the well-being of medical students. PMID- 15795411 TI - The exaggerated relations between diet, body weight and mortality: the case for a categorical data approach. PMID- 15795413 TI - Thromboembolic events in gastric cancer: high incidence in patients receiving irinotecan- and bevacizumab-based therapy. PMID- 15795412 TI - Nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease is emerging as the most common chronic liver condition in the Western world. It is associated with insulin resistance and frequently occurs with features of the metabolic syndrome. Disease presentation ranges from asymptomatic elevated liver enzyme levels to cirrhosis with complications of liver failure and hepatocellular carcinoma. Current treatment recommendations are limited to weight loss and exercise, although several promising medications are on the horizon. In this article we discuss the etiology, pathogenesis and diagnosis of nonalcoholic fatty liver disease as well as approaches to its management. PMID- 15795414 TI - Contralateral prophylactic mastectomy: quantifying benefits and weighing the harms. PMID- 15795415 TI - Efficacy of prophylactic mastectomy in women with unilateral breast cancer: a cancer research network project. AB - PURPOSE: We investigated the efficacy of contralateral prophylactic mastectomy (CPM) in reducing contralateral breast cancer incidence and breast cancer mortality among women who have already been diagnosed with breast cancer. METHODS: This retrospective cohort study comprised approximately 50,000 women who were diagnosed with unilateral breast cancer during 1979 to 1999. Using computerized data confirmed by chart review, we identified 1,072 women (1.9%) who had CPM. We obtained covariate information for these women and for a sample of 317 women who did not undergo CPM. RESULTS: The median time from initial breast cancer diagnosis to the end of follow-up was 5.7 years. Contralateral breast cancer developed in 0.5% of women with CPM, metastatic disease developed in 10.5%, and subsequent breast cancer developed in 12.4%; 8.1% died from breast cancer. Contralateral breast cancer developed in 2.7% of women without CPM, and 11.7% died of breast cancer. After adjustment for initial breast cancer characteristics, treatment, and breast cancer risk factors, the hazard ratio (HR) for the occurrence of contralateral breast cancer after CPM was 0.03 (95% CI, 0.006 to 0.13). After adjustment for breast cancer characteristics and treatment, the HRs for the relationship of CPM with death from breast cancer, with death from other causes, and with all-cause mortality were 0.57 (95% CI, 0.45 to 0.72), 0.78 (95% CI, 0.57 to 1.06), and 0.60 (95% CI, 0.50 to 0.72), respectively. CONCLUSION: CPM seems to protect against the development of contralateral breast cancer, and although women who underwent CPM had relatively low all-cause mortality, CPM also was associated with decreased breast cancer mortality. PMID- 15795416 TI - Cigarette smoke alters human vitamin E requirements. AB - Vitamin E is a lipophilic chain-breaking antioxidant that prevents lipid peroxidation. Although cigarette smoke is a potent source of oxidative stress that depletes vitamin E in vitro, it is unclear whether it has a similar effect in vivo, particularly in humans. Therefore, this review will discuss the role of cigarette smoke on gamma-tocopherol (gamma-T) nitration, its effect on alpha tocopherol (alpha-T) biokinetics in smokers, and the changes in the synthesis, plasma concentrations, and urinary excretion of the vitamin E metabolite (CEHC; carboxy-ethyl-hydroxy-chroman). Last, the possibility of CEHC as a biomarker of vitamin E status will be assessed as will the question whether smokers have increased dietary requirements of vitamin E. PMID- 15795417 TI - David Marine and the problem of goiter. PMID- 15795418 TI - Sexual dimorphism in human lipid metabolism. AB - The existing work demonstrates that striking differences exist between men and women in lipid kinetics. These differences cannot be explained simply by the presence and action of sex hormones and are not always due to secondary, phenotypic traits that characterize men and women (e.g., body-composition, regional fat distribution). In fact, some of these secondary traits may even be the result of sexual dimorphism in metabolism, and being of female or male genotype also determines intermediary metabolism. This review provides an overview of the currently available information regarding sexual dimorphism in human lipid metabolism but does not provide an in-depth account of current knowledge (due to limited space); it will be a broad introduction to those interested in the field and will, hopefully, stimulate further efforts to unravel the secrets of male and female metabolism. What has been discovered so far regarding differences in lipid metabolism between men and women is likely only the tip of the iceberg; clearly, more work is necessary to fully understand human substrate metabolism and the implications the presence of sexual dimorphism in the control of substrate kinetics has on the prevention and treatment of disease. PMID- 15795419 TI - Dietary olive oil supplemented with fish oil, rich in EPA and DHA (n-3) polyunsaturated fatty acids, attenuates colonic inflammation in rats with DSS induced colitis. AB - Previous studies proposed a protective role of the dietary intake of (n-3) PUFA in human inflammatory bowel disease (IBD), but almost no studies have been performed using olive oil. The aims of the present study were to test the beneficial effects of an olive oil-based diet with or without fish oil, rich in (n-3) PUFA, in the dextran sodium sulfate (DSS) model of rat colitis and to elucidate the mechanisms involved in their potential beneficial effects, with special attention to the production of some of the mediators involved in the intestinal inflammatory response, such as leukotriene B(4) (LTB(4)), tumor necrosis factor alpha (TNFalpha) and nitric oxide (NO). Rats were fed the different diets for 2 wk before colitis induction and thereafter until colonic evaluation 15 d later. Colitic rats fed the olive oil-based diet had a lower colonic inflammatory response than those fed the soybean oil diet, and this beneficial effect was increased by the dietary incorporation of (n-3) PUFA. A restoration of colonic glutathione levels and lower colonic NO synthase expression occurred in all colitic rats fed an olive oil diet compared with the control colitic group that consumed the soybean oil diet. However, (n-3) PUFA incorporation into an olive oil diet significantly decreased colonic TNFalpha and LTB(4) levels compared with colitic rats that were not supplemented with fish oil. These results affirm the benefits of an olive oil diet in the management of IBD, which are further enhanced by the addition of (n-3) PUFA. PMID- 15795420 TI - Vitamin A deficiency alters the structure and collagen IV composition of rat renal basement membranes. AB - Retinoids can modulate the expression of extracellular matrix (ECM) proteins with variable results depending on other contributing factors. Because changes in these proteins may alter the composition and impair the function of specialized ECM structures such as basement membranes (BMs), we studied the effects of vitamin A deficiency on renal BMs during the growing period. Newborn male rats were fed a vitamin A-deficient (VAD) diet for 50 d. The ultrastructure of renal BMs was analyzed by electron microscopy. Total collagen IV, the different alpha(IV) chains, matrix degrading metalloproteinases (MMP), and tissue inhibitors of metalloproteinases (TIMP) were quantified by immunocytochemistry and/or Western blotting. Tumor necrosis factor-alpha and interleukin-1beta were measured by ELISA. Semiquantitative RT-PCR was used for determining the steady state levels for each alpha(IV) chain mRNA. VAD renal BMs showed an irregular thickening, particularly tubular BM. The total collagen IV content was increased, but there was a differential expression of the collagen IV chains. The protein amounts for alpha1(IV), alpha4(IV), and alpha5(IV) were similarly increased, whereas alpha2(IV) and alpha3(IV) were decreased. The levels of mRNA for each collagen IV chain changed in parallel with those of the corresponding protein. Both MMP2 and MMP9 were diminished, but no change was detected in TIMP1 or TIMP2. Our data indicate that nutritional VAD leads to alterations in the structure of renal BMs and to quantitative and qualitative variations in its collagen IV composition. These changes may be a factor predisposing to or resulting in kidney malfunction and renal disease. PMID- 15795421 TI - Microsomal triglyceride transfer protein gene expression and ApoB secretion are inhibited by bitter melon in HepG2 cells. AB - Momordica charantia or bitter melon is traditionally used as an antidiabetic agent in Asia, Africa, and South America. Recent studies indicate that bitter melon can also lower plasma lipids and VLDL in diabetic animal models as well as animals fed a high-fat diet, suggesting an effect on lipoprotein metabolism. The aim of this study was to delineate the cellular and molecular mechanisms involved in the lipid-lowering properties of bitter melon and regulation of apolipoprotein B (apoB). Human hepatoma cells, HepG2, treated with bitter melon juice (BMJ) for 24 h reduced apoB secretion with and without the addition of lipids (P < 0.05). However, BMJ did not increase apoB secretion in cells treated with N-acetyl leucyl-leucyl-norleucinal, indicating a lack of effect on the proteasomal degradation pathway. BMJ reduced the secretion of new triglycerides (P < 0.05) and decreased microsomal triglyceride transfer protein (MTP) mRNA expression, suggesting that lipid bioavailability and lipidation of lipoprotein assembly are likely involved in decreased apoB secretion. Interestingly, BMJ increased the nuclear translocation of the mature form of sterol regulatory element-binding protein-1c (SREBP-1c, P < 0.05), involved in MTP secretion. Our data suggest that BMJ is a potent inhibitor of apoB secretion and TG synthesis and secretion that may be involved in the plasma lipid- and VLDL-lowering effects observed in animal studies. PMID- 15795422 TI - (-)Epigallocatechin gallate and quercetin enhance survival signaling in response to oxidant-induced human endothelial apoptosis. AB - We reported recently that (-)epigallocatechin gallate and quercetin inhibited H2O2-induced apoptosis through modulation of the expression of apoptosis-related Bcl-2 and Bax in endothelial cells. This study attempted to identify possible regulatory sites and mechanisms of antiapoptotic flavonoids, focusing on ROS mediated signaling in HUVEC. The effects of apigenin on the signaling pathway downstream were compared. Submillimolar H2O2 caused >30% cell killing with intracellular oxidant generation. H2O2-induced oxidant generation markedly decreased total intracellular glutathione (GSH) levels. Micromolar ( )epigallocatechin gallate and quercetin partially eliminated the dichlorodihydrofluorescein (DCF) and phospho-p53 staining, suggesting that these flavonoids inhibited the accumulation of intracellular oxidants and nuclear transactivation of p53 in H2O2-exposed cells. In contrast, cells treated with apigenin remained DCF and phospho-p53 staining positive in response to H2O2. ( )Epigallocatechin gallate significantly raised the total GSH level that had been depleted by H2O2. Caspase-3 activity was enhanced by H2O2, and this increase was inhibited by (-)epigallocatechin gallate and quercetin. Additionally, the upregulation of caspase-3 activation was reversed by these flavonoids at > or =10 micromol/L; these inhibitory effects were dose dependent. Western blot data revealed that H2O2 upregulated phosphorylation of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK), which was rapidly reversed by quercetin within 30 min; H2O2 activation of c-Jun was downregulated. ( )Epigallocatechin gallate inhibited H2O2-induced phosphorylation of JNK and p38 MAPK after 60 min. These results reveal that quercetin blocks JNK- and p38 MAPK related signaling triggered by the oxidant and may regulate expression of apoptotic downstream genes, preventing apoptosis and promoting cell survival. ( )Epigallocatechin gallate may function as an antiapoptotic agent through other antiapoptotic pathways. PMID- 15795423 TI - Dietary L-arginine supplementation reduces fat mass in Zucker diabetic fatty rats. AB - This study was conducted to test the hypothesis that dietary supplementation of arginine, the physiologic precursor of nitric oxide (NO), reduces fat mass in the Zucker diabetic fatty (ZDF) rat, a genetically obese animal model of type-II diabetes mellitus. Male ZDF rats, 9 wk old, were pair-fed Purina 5008 diet and received drinking water containing arginine-HCl (1.51%) or alanine (2.55%, isonitrogenous control) for 10 wk. Serum concentrations of arginine and NO(x) (oxidation products of NO) were 261 and 70% higher, respectively, in arginine supplemented rats than in control rats. The body weights of arginine-treated rats were 6, 10, and 16% lower at wk 4, 7, and 10 after the treatment initiation, respectively, compared with control rats. Arginine supplementation reduced the weight of abdominal (retroperitoneal) and epididymal adipose tissues (45 and 25%, respectively) as well as serum concentrations of glucose (25%), triglycerides (23%), FFA (27%), homocysteine (26%), dimethylarginines (18-21%), and leptin (32%). The arginine treatment enhanced NO production (71-85%), lipolysis (22 24%), and the oxidation of glucose (34-36%) and octanoate (40-43%) in abdominal and epididymal adipose tissues. Results of the microarray analysis indicated that arginine supplementation increased adipose tissue expression of key genes responsible for fatty acid and glucose oxidation: NO synthase-1 (145%), heme oxygenase-3 (789%), AMP-activated protein kinase (123%), and peroxisome proliferator-activated receptor gamma coactivator-1alpha (500%). The induction of these genes was verified by real-time RT-PCR analysis. In sum, arginine treatment may provide a potentially novel and useful means to enhance NO synthesis and reduce fat mass in obese subjects with type-II diabetes mellitus. PMID- 15795425 TI - Mulberry (Morus alba L.) leaves and their major flavonol quercetin 3-(6 malonylglucoside) attenuate atherosclerotic lesion development in LDL receptor deficient mice. AB - The effects of dietary consumption of mulberry (Morus alba L.) leaves and their major flavonol glycoside, quercetin 3-(6-malonylglucoside) (Q3MG), on the development of atherosclerotic lesions, in relation to the susceptibility of plasma LDL to oxidative modification, was studied in LDL receptor-deficient (LDLR /-) mice. Male mice aged 8 wk were randomly assigned to 4 groups (control, quercetin, Q3MG, and mulberry). The control group was fed an atherogenic-diet containing 3 g cholesterol and 15 g cocoa butter/100 g. The other experimental groups were fed the same atherogenic diet supplemented with 0.05 g quercetin/100 g for the quercetin group, 0.05 g Q3MG/100 g for the Q3MG group, and 3 g dried mulberry-leaf powder/100 g for the mulberry group. The mice were fed their respective diets for 8 wk. The susceptibility of LDL to oxidative modification was significantly decreased in the Q3MG- and mulberry-treated mice, as evidenced by the 44.3 and 42.2% prolongation of the lag phase for conjugated diene formation compared with that of the control mice. The atherosclerotic lesion area in both the Q3MG- and mulberry-treated mice was significantly reduced by 52% compared with that of the controls. However, in the quercetin group, no protective effects were observed against LDL oxidation or atherosclerotic lesion formation. In conclusion, mulberry leaves attenuated the atherosclerotic lesion development in LDLR-/- mice through enhancement of LDL resistance to oxidative modification, and these antioxidative and antiatherogenic protective effects were attributed mainly to Q3MG, the quantitatively major flavonol glycoside in mulberry leaves. PMID- 15795424 TI - Grape powder polyphenols attenuate atherosclerosis development in apolipoprotein E deficient (E0) mice and reduce macrophage atherogenicity. AB - The beneficial health effects of red wine have been attributed to the antioxidant activity of its polyphenols. The present study investigated the effects of a standardized freeze-dried powder made from fresh grapes, rich in grape-specific polyphenols and free of alcohol, on oxidative stress, atherogenicity of macrophages, and the development of atherosclerotic lesions in apolipoprotein E deficient (E(0)) mice. Thirty E(0) mice were assigned to 3 groups. Mice consumed water alone (control), 150 mug total polyphenols/d in the form of grape powder (grape powder), or the equivalent amount of glucose and fructose (placebo) in drinking water for 10 wk. Consumption of grape powder reduced the atherosclerotic lesion area by 41% (P < 0.0002) compared to the control or placebo mice. The antiatherosclerotic effect was at least partly due to a significant 8% reduction in serum oxidative stress, an up to 22% increase in serum antioxidant capacity, a significant 33% reduction in macrophage uptake of oxidized LDL, and a 25% decrease in macrophage-mediated oxidation of LDL relative to controls. Grape powder directly protected both plasma LDL and macrophages from oxidative stress in vitro. We conclude that polyphenols from fresh grape powder directly affect macrophage atherogenicity by reducing macrophage-mediated oxidation of LDL and cellular uptake of oxidized LDL. Both of these processes can eventually reduce macrophage cholesterol accumulation and foam cell formation and hence attenuate atherosclerosis development. PMID- 15795426 TI - The lowering of plasma lipids following a weight reduction program is related to increased expression of the LDL receptor and lipoprotein lipase. AB - To determine whether changes in plasma lipids following a weight loss program were related to modifications in gene expression of the LDL receptor (LDL-R), lipoprotein lipase (LPL), and 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, overweight/obese premenopausal women were recruited. The 10-wk, randomized, double-blind intervention consisted of a hypoenergetic diet, high in protein (30% energy) and low in carbohydrate (40% energy), increased physical activity (number of steps taken per day), and intake of a supplement (carnitine or placebo). Our initial hypothesis was that carnitine would enhance the beneficial effects of weight loss on plasma lipids and anthropometrics. Because the carnitine and placebo groups did not differ in any of the measured variables, data for all subjects were pooled and comparisons were made between baseline and postintervention. Mean weight loss was 4.4 kg (P < 0.001), and plasma triglycerides (TG), total, and LDL cholesterol (LDL-C) were reduced by 31.8, 9.9, and 11.9%, respectively (P < 0.001). The expression of the genes of interest was measured in RNA extracted from mononuclear cells at baseline and postintervention using a semiquantitative RT-PCR method. Glyceraldehyde-3-phosphate dehydrogenase was used as an internal control. After 10 wk, there was a 25.7% increase in the abundance of LPL mRNA (P < 0.01) and a 27.7% increase in that of LDL-R mRNA (P < 0.01). The expression of HMG-CoA reductase was not altered by weight loss. The results suggest that the increased expression of the LDL-R and LPL after the intervention might have contributed to the lower plasma LDL-C and TG observed in these women. PMID- 15795427 TI - Epithelium-specific glutathione peroxidase, Gpx2, is involved in the prevention of intestinal inflammation in selenium-deficient mice. AB - Mice deficient in 2 intracellular selenium (Se)-dependent glutathione peroxidases (Gpx1 and Gpx2), by genetically disrupting both alleles of the Gpx1 and Gpx2 genes (Gpx1(-/-)Gpx2(-/-)), develop ileocolitis around weaning. However, decreased Gpx activity in Se-depleted wild-type animals does not produce pathology in the gastrointestinal tract. Because a small percentage of Se sufficient Gpx1(+/-)Gpx2(-/-) mice have mild ileocolitis, we hypothesized that Se deficient Gpx1(+/-)Gpx2(-/-) mice will develop severe ileocolitis similarly to the Gpx1(-/-)Gpx2(-/-) mice, and even a trace amount of Gpx2 can protect intestinal mucosa against inflammation. To test our hypothesis, we fed mice at various stages of development with either Gpx1(+/)(-)Gpx2(-/-) or Gpx1(-/ )Gpx2(+/)(-) genotypes an Se-deficient diet for 4-5 wk and assessed the symptoms and pathology. Gpx1(+/)(-)Gpx2(-/-) mice that were deprived of Se in utero or at weaning (18-22 d of age), but not as young adults (31-51 d of age), manifested significantly worse pathology than their Se-sufficient counterparts. Both Gpx1 and Gpx2 activities and mRNA levels were significantly depressed in the ileum of Se-deprived mice. In mice deprived in utero, the pathology included acute inflammation with neutrophil and monocyte infiltration particularly in the colon and was externally manifested by perianal alopecia and ulceration. On the other hand, Gpx1(-/-)Gpx2(+/)(-) mice were unaffected by Se deprivation, regardless of the age of onset. The results show that a trace amount of Gpx2 is protective against ileocolitis, and Se-deficient young Gpx1(+/-)Gpx2(-/-) mice will develop pathology and symptoms similar to Se-adequate Gpx1(-/-)Gpx2(-/-) mice. PMID- 15795428 TI - Dietary retinoic acid alters vitamin A kinetics in both the whole body and in specific organs of rats with low vitamin A status. AB - To study the effects of exogenous retinoic acid on vitamin A (VA) metabolism, we analyzed previously collected tracer kinetic data on VA dynamics in rats with low vitamin A (LA) status either with (LA+RA) or without (LA) retinoic acid supplementation. In spite of low VA intake ( approximately 7 nmol/d), the LA+RA rats were in a slight positive VA balance (0.325 nmol/d vs. -0.168 for LA) for 35 d after administration of [(3)H]retinol-labeled plasma. Using the Windows version of the Simulation, Analysis and Modeling software, we determined that the VA disposal rate was lower in LA+RA than in LA rats (3.98 vs. 5.00 nmol/d) as was the system fractional catabolic rate (0.0548 vs. 0.110 d(-1)). Model-predicted traced mass and residence times (the average time that a molecule of retinol spends in an organ before irreversible loss) were higher for liver (19.4 vs. 1.8 nmol; 5.0 vs. 0.36 d), kidneys (7.0 vs. 2.1 nmol; 1.4 vs. 0.42 d), small intestine (2.1 vs. 0.42 nmol; 0.43 vs. 0.084 d), and lungs (3.2 vs. 0.10 nmol; 1.6 vs. 0.021 d) in the LA+RA compared with the LA rats; there were no major differences for eyes, testes, adrenal glands, or remaining carcass. We conclude that RA supplementation of rats with low VA status affects VA metabolism at both the whole-body level and in specific organs. These organs (liver, kidneys, small intestine, and lungs) have the enzymatic capability and an appropriate cell type to store retinyl esters. PMID- 15795429 TI - Dietary L-carnitine enhances the lymphatic absorption of fat and alpha-tocopherol in ovariectomized rats. AB - This study was conducted to determine whether the feeding of dietary L-carnitine (CN) improves the intestinal absorption of fat and alpha-tocopherol (alphaTOH) in ovariectomized (OX) rats. OX adult rats were weight-matched and assigned to 2 groups fed a modified AIN-93G diet containing alphaTOH-stripped soybean oil without (-CN) or with (+CN) supplemental CN at 150 mg/kg diet. At 5 wk, each rat with a lymph cannula was infused intraduodenally at 3.0 mL/h with a lipid emulsion consisting of 565 micromol triolein labeled with (14)C ((14)C-OA), 3.6 micromol alphaTOH, and 396 micromol sodium taurocholate in 24 mL PBS buffer. Lymph was collected hourly for 8 h and analyzed for lipids. The lymphatic absorption of alphaTOH for 8 h in +CN rats (899 +/- 201 nmol) was higher (P < 0.05) than in -CN rats (587 +/- 92 nmol). The absorption of (14)C-OA in +CN rats (53.5 +/- 4.0% dose/8 h) also was increased (P < 0.05) compared with -CN rats (47.6 +/- 5.0% dose/8 h). Lymph flow did not differ between the groups. When bile was diverted but with infusion of sodium taurocholate, the lymphatic absorption of lipids did not differ. The present study provides evidence that dietary CN enhances the rates and amounts of lymphatic absorption of alphaTOH and fat in OX rats. Our findings suggest that dietary CN may influence the process of lipid packaging and absorption by the enterocyte in OX rats, and may explain in part the increased status of alphaTOH in CN-fed animals. PMID- 15795430 TI - trans-Resveratrol, a natural antioxidant from grapes, increases sperm output in healthy rats. AB - trans-Resveratrol was reported to have health benefits including anticarcinogenic effects and protection against cardiovascular disease. One of the mechanisms by which it exerts its action is through modulating the estrogen response systems. Because estrogen is involved in male reproductive biology, we investigated the effect of trans-resveratrol on testis and spermatogenesis. Adult male rats were divided into 2 groups. The treated group was administered by gavage 20 mg/(kg . d) of trans-resveratrol suspended in 10 g/L of carboxymethylcellulose for 90 d, whereas the control group received only carboxymethylcellulose during the same period. The relative weight of testes did not differ between the groups. However, the diameter of the seminiferous tubules was significantly reduced from 437.5 +/- 0.1 mum in the controls to 310.9 +/- 0.1 mum in the resveratrol-treated rats. This decrease was accompanied by a significant increase in tubular density, from 3.20 +/- 0.18 in controls to 6.58 +/- 0.18 tubules/mm(2) in the treated group. Moreover, sperm counts were significantly greater in the resveratrol-treated rats (24.8 +/- 3.30 x 10(7)) than in the control group (14.1 +/- 0.80 x 10(7)), but sperm quality did not differ. Serum concentrations of gonadotrophins and testosterone were significantly higher in the resveratrol-treated group. We identified a novel activity of trans-resveratrol. The daily oral administration of this phytochemical to adult male rats enhanced sperm production by stimulating the hypothalamic-pituitary-gonadal axis, without inducing adverse effects. PMID- 15795431 TI - Carnitine deficiency and supplementation do not affect the gene expression of carnitine biosynthetic enzymes in rats. AB - Starved male weanling rats supplemented with 20 mmol/L pivalate in their drinking water exhibit significantly depressed concentrations of carnitine in tissues and plasma. In addition, pivalate supplementation has been linked with increased renal and hepatic trimethyllysine hydroxylase (TMLH) activity, whereas carnitine supplementation has been associated with significantly decreased hepatic gamma butyrobetaine hydroxylase (BBH) activity. The purpose of this study was to determine whether pivalate or carnitine supplementation affects the activity and genetic expression of 2 enzymes of carnitine (Cn) biosynthesis, TMLH and BBH, expressed as mRNA abundance, relative to the abundance of beta-actin mRNA. Male weanling rats were administered the control treatment (C; n = 6), the pivalate treatment (P; n = 7), or the pivalate treatment plus supplemental dietary carnitine (P+Cn; n = 7). Rats in group P had elevated renal TMLH activity, relative to the other groups (P < 0.05). The groups did not differ in the abundance of renal or hepatic TMLH or BBH mRNA. A previously unreported finding was the quantifiable level of renal BBH mRNA, which was verified by direct sequencing of the BBH cDNA product amplified from kidney RNA. The groups did not differ in renal BBH mRNA abundance and renal BBH enzyme activity was not detected. Thus, the alterations in enzyme activities in the pivalate-treated rats are not regulated at the transcriptional level, and are apparently related to post-transcriptional effects on the enzymes themselves. PMID- 15795432 TI - Threonine utilization is high in the intestine of piglets. AB - The whole-body threonine requirement in parenterally fed piglets is substantially lower than that in enterally fed piglets, indicating that enteral nutrition induces intestinal processes in demand of threonine. We hypothesized that the percentage of threonine utilization for oxidation and intestinal protein synthesis by the portal-drained viscera (PDV) increases when dietary protein intake is reduced. Piglets (n = 18) received isocaloric normal or protein restricted diets. After 7 h of enteral feeding, total threonine utilization, incorporation into intestinal tissue, and oxidation by the PDV, were determined with stable isotope methodology [U-(13)C threonine infusion]. Although the absolute amount of systemic and dietary threonine utilized by the PDV was reduced in protein-restricted piglets, the percentage of dietary threonine intake utilized by the PDV did not differ between groups (normal protein 91% vs. low protein 85%). The incorporation of dietary threonine into the proximal jejunum was significantly different compared with the other intestinal segments. Dietary, rather than systemic threonine was preferentially utilized for protein synthesis in the small intestinal mucosa in piglets that consumed the normal protein diet (P < 0.05). Threonine oxidation by the PDV was limited during normal protein feeding. In protein-restricted pigs, half of the total whole-body oxidation occurred in the PDV. We conclude that, in vivo, the PDV have a high obligatory visceral requirement for threonine. The high rate of intestinal threonine utilization is due mainly to incorporation into mucosal proteins. PMID- 15795433 TI - Absorption and loss of iron in toddlers are highly correlated. AB - For estimating the requirements for dietary iron, it is important to know the amount of iron that is lost from the body. Inevitable losses of iron have been determined in adult humans but not in infants or children. We administered (58)Fe, the least abundant stable isotope of iron, to free-living infants at 168 d of age (5.6 mo) and followed them to age 26 mo. There was no dietary restriction after isotope administration. Blood was obtained at regular intervals for determination of isotopic enrichment and indices of iron status. We estimated the quantity of circulating iron, noncirculating active iron, and storage iron at each age. The administered isotope equilibrated with total body iron by 13 mo of age. From 13 to 26 mo of age, we estimated inevitable loss and absorption of iron from the change in tracer abundance in circulating iron. The rate of decrease of tracer abundance was proportional to addition of tracee, i.e., absorption of iron. Conversely, the rate of decrease in quantity of tracer was proportional to removal of tracee, i.e., loss of iron. From 13 to 26 mo of age, iron absorption was (mean +/- SD) 0.49 +/- 0.13 mg/d and inevitable iron loss was 0.25 +/- 0.12 mg/d. Intersubject variability of iron loss and iron absorption was high, and iron loss and absorption were highly correlated (r = 0.789, P < 0.001). Iron stores were low throughout the study and decreased significantly from 13 to 26 mo of age, suggesting that iron absorption from the diet was inadequate to maintain or increase iron nutritional status. The data suggest that, in this cohort, which may be representative, the intake of bioavailable iron from 13 to 26 mo of age was insufficient to maintain iron nutritional status. PMID- 15795434 TI - Supplementation with conjugated linoleic acid for 24 months is well tolerated by and reduces body fat mass in healthy, overweight humans. AB - After 12 mo in a randomized, double-blind, placebo-controlled trial of conjugated linoleic acid (CLA) supplementation (2 groups received CLA as part of a triglyceride or as the free fatty acid, and 1 group received olive oil as placebo), 134 of the 157 participants who concluded the study were included in an open study for another 12 mo. The goals of the extension study were to evaluate the safety [with clinical chemistry analyses and reported adverse events (AEs)] and assess the effects of CLA on body composition [body fat mass (BFM), lean body mass (LBM), bone mineral mass (BMM)], body weight, and BMI. All subjects were supplemented with 3.4g CLA/d in the triglyceride form. Circulating lipoprotein(a) and thrombocytes increased in all groups. There was no change in fasting blood glucose. Aspartate amino transferase, but not alanine amino transferase, increased significantly. Plasma total cholesterol and LDL cholesterol were reduced, whereas HDL cholesterol and triglycerides were unchanged. The AE rate decreased compared with the first 12 mo of the study. Body weight and BFM were reduced in the subjects administered the placebo during the initial 12 mo study ( 1.6 +/- 3.2 and -1.7 +/- 2.8 kg, respectively). No fat or body weight changes occurred in the 2 groups given CLA during the initial 12 mo. LBM and BMM were not affected in any of the groups. Changes in body composition were not related to diet and/or training. In conclusion, this study shows that CLA supplementation for 24 mo in healthy, overweight adults was well tolerated. It confirms also that CLA decreases BFM in overweight humans, and may help maintain initial reductions in BFM and weight in the long term. PMID- 15795435 TI - Coffee oil consumption increases plasma levels of 7alpha-hydroxy-4-cholesten-3 one in humans. AB - Unfiltered coffee brews such as French press and espresso contain a lipid from coffee beans named cafestol that raises serum cholesterol in humans. Cafestol decreases the expression and activity of cholesterol 7alpha-hydroxylase, the rate limiting enzyme in the classical pathway of bile acid synthesis, in cultured rat hepatocytes and livers of APOE3Leiden mice. Inhibition of bile acid synthesis has been suggested to be responsible for the cholesterol-raising effect of cafestol. Therefore, we assessed whether cafestol decreases the activity of cholesterol 7alpha-hydroxylase in humans. Because liver biopsies were not feasible, we measured plasma levels of 7alpha-hydroxy-4-cholesten-3-one, a marker for the activity of cholesterol 7alpha-hydroxylase in the liver. Plasma 7alpha-hydroxy-4 cholesten-3-one was measured in 2 separate periods in which healthy volunteers consumed coffee oil containing cafestol (69 mg/d) for 5 wk. Plasma levels of 7alpha-hydroxy-4-cholesten-3-one increased by 47 +/- 13% (mean +/- SEM, n = 38, P = 0.001) in the first period and by 23 +/- 10% (n = 31, P = 0.03) in the second treatment period. Serum cholesterol was raised by 23 +/- 2% (P < 0.001) in the first period and by 18 +/- 2% (P < 0.001) in the second period. We corrected individual 7alpha-hydroxy-4-cholesten-3-one levels for serum cholesterol levels, because coffee oil increases serum cholesterol and 7alpha-hydroxy-4-cholesten-3 one is probably present in the lipoprotein fraction of serum. After correction, the increase in 7alpha-hydroxy-4-cholesten-3-one was 24 +/- 11% (P = 0.04) in the first period and there was no effect in period 2. Our study showed that coffee oil did not decrease, and actually increased, plasma levels of 7alpha-hydroxy-4 cholesten-3-one in humans in 2 separate treatment periods. Therefore, this study does not support the hypothesis that cafestol decreases bile acid synthesis in humans. PMID- 15795436 TI - Enrichment of tomato paste with 6% tomato peel increases lycopene and beta carotene bioavailability in men. AB - A high intake of tomato products is associated with a lower incidence of upper aerodigestive tract and prostate cancers. This beneficial effect might be explained by a higher intake of carotenoids such as lycopene and/or beta carotene. Because tomato peels, usually eliminated during tomato processing, are a valuable source of these carotenoids, we designed a study to examine whether a tomato paste enriched in tomato peels (ETP, 6% peel) increases the absorption of these carotenoids compared to a classically made tomato paste (CTP). Carotenoid bioaccessibility was evaluated using an in vitro digestion model by measuring the amount of carotenoids transferred from the pastes to micelles. Carotenoid absorption by human intestinal cells (Caco-2) was evaluated after the addition of carotenoid-rich micelles (obtained from the in vitro digestion of the 2 pastes). Carotenoid bioavailability in humans was assessed by measuring chylomicron carotenoid responses in a postprandial experiment in which 8 healthy men consumed 2 meals containing either the ETP or the CTP. ETP contained 47.6 mg lycopene (58% more than CTP) and 1.75 mg beta-carotene (99% more than CTP) per 100 g of paste. In micelles, 30% more lycopene and 81% more beta-carotene were recovered after ETP than after CTP in vitro digestion. The amount of carotenoids absorbed by Caco 2 cells was 75% greater (P < or = 0.05) for lycopene and 41% greater (P < or = 0.05) for beta-carotene after the addition of micelles from ETP than from CTP. After ETP intake the chylomicron beta-carotene response was 74% greater than after CTP intake, and the lycopene response tended to be greater (34.1%, P = 0.093). Peel enrichment of tomato paste with tomato peel is an interesting option for increasing lycopene and beta-carotene intakes. PMID- 15795437 TI - Pharmacokinetics of enterolignans in healthy men and women consuming a single dose of secoisolariciresinol diglucoside. AB - High concentrations of enterolignans in plasma are associated with a lower risk of acute coronary events. However, little is known about the absorption and excretion of enterolignans. The pharmacokinetic parameters and urinary excretion of enterodiol and enterolactone were evaluated after consumption of their purified plant precursor, secoisolariciresinol diglucoside (SDG). Twelve healthy volunteers ingested a single dose of purified SDG (1.31 micromol/kg body wt). Enterolignans appeared in plasma 8-10 h after ingestion of the purified SDG. Enterodiol reached its maximum plasma concentration 14.8 +/- 5.1 h (mean +/- SD) after ingestion of SDG, whereas enterolactone reached its maximum 19.7 +/- 6.2 h after ingestion. The mean elimination half-life of enterodiol (4.4 +/- 1.3 h) was shorter than that of enterolactone (12.6 +/- 5.6 h). The mean area under the curve of enterolactone (1762 +/- 1117 nmol/L . h) was twice as large as that of enterodiol (966 +/- 639 nmol/L . h). The mean residence time for enterodiol was 20.6 +/- 5.9 h and that for enterolactone was 35.8 +/- 10.6 h. Within 3 d, up to 40% of the ingested SDG was excreted as enterolignans via urine, with the majority (58%) as enterolactone. In conclusion, a substantial part of enterolignans becomes available in the blood circulation and is subsequently excreted. The measured mean residence times and elimination half-lives indicate that enterolignans accumulate in plasma when consumed 2-3 times a day and reach steady state. Therefore, plasma enterolignan concentrations are expected to be good biomarkers of dietary lignan exposure and can be used to evaluate the effects of lignans. PMID- 15795438 TI - Plasma zinc concentrations are depressed during the acute phase response in children with falciparum malaria. AB - Plasma concentrations of some micronutrients are altered in the setting of acute infectious or inflammatory stress. Previous studies have provided conflicting evidence concerning the extent and direction of changes in plasma zinc concentrations during the acute phase response. We carried out an observational cohort study in 689 children enrolled in a randomized trial of zinc supplementation during acute falciparum malaria in order to evaluate the relation between plasma zinc concentration and the acute phase response. Plasma zinc was measured by atomic absorption spectrophotometry. On admission, 70% of all subjects had low plasma zinc (<9.2 micromol/L). Multivariate analysis of predictors of admission plasma zinc showed that admission C-reactive protein (CRP), parasite density, and study site were the most important predictors. Predictors of changes in plasma zinc from admission to 72 h included baseline CRP, change in CRP, treatment group, study site, and baseline zinc concentration. In children with acute malaria infection, baseline plasma zinc concentrations were very low and were inversely correlated with CRP (r = -0.24, P < 0.0001) and the degree of parasitemia (r = -0.19, P < 0.0001). Even when CRP and time were taken into account, zinc supplementation increased plasma zinc concentration from admission to 72 h. When available, plasma zinc concentrations should be interpreted with concurrent measures of the acute phase response such as CRP. In children whose age, diet, and/or nutritional status place them at risk of zinc deficiency, those with low plasma zinc levels should be supplemented with oral zinc and followed for clinical and/or biochemical response. PMID- 15795439 TI - Iron and vitamin A deficiency in long-term African refugees. AB - Five cross-sectional surveys were conducted in African refugee camps to assess the level of iron deficiency anemia and vitamin A deficiency in populations dependent on long-term international food aid and humanitarian assistance. The prevalence of anemia in children [hemoglobin (Hb) <110 g/L] was high, with >60% affected in 3 of 5 camps. Iron deficiency [serum transferrin receptor (sTfR) >8.5 mg/L] was also high, ranging from 23 to 75%; there was also a strong ecological correlation between the prevalence of iron deficiency and anemia among different camps. Within camps, sTfR predicted the concentration of Hb with adjusted R(2) values ranging from 0.19 to 0.51. Although children were more affected, anemia was also a public health problem in adolescents and women. The effect of recent recommendations on Hb cutoff values for African populations was assessed and found to produce decreases in the prevalence of anemia of between 5 and 21%; this did not affect the public health categorization of the anemia problem within the most affected camps. Mean serum retinol in children, after adjustment for infection status, ranged from 0.72 +/- 0.2 to 0.88 +/- 0.2 micromol/L in the 4 camps assessed and vitamin A deficiency (<0.7 micromol/L) was present at levels ranging from 20.5 to 61.7%. In areas in which vitamin A capsule distribution programs were in effect, coverage ranged from 3.5 up to 66.2%. The high level of micronutrient deficiencies seen in long-term refugees argues in favor of further enhancements in food aid fortification and the strengthening of nutrition and public health programs. PMID- 15795440 TI - Iron deficiency and physical growth predict attainment of walking but not crawling in poorly nourished Zanzibari infants. AB - Locomotion allows infants to explore their environment, promoting development in other domains. Motor progression involves biological systems and experiential factors. Nutritional deficiencies could interfere with systems involved in locomotion. This study examined the associations between height-for-age (HAZ), weight-for-height (WHZ) Z-scores and anemia-iron status on locomotion in 646 Zanzibari infants. Motor milestones were assessed by trained observers using a 14 item scale. Two mutually exclusive samples were created. The crawling sample (n = 167, 6-18 mo old) included infants that crawled only or did not crawl; the walking sample (n = 479, 9-18 mo old) included children that walked alone or did not walk alone. Of the crawling and walking samples, 82.6 and 83.9% respectively, were iron deficient and/or anemic (hemoglobin < 100 g/L; zinc protoporphyrin > or = 90 micromol/mol heme). Stunting (HAZ less than -2) occurred in 30.5% of the crawling sample and 38.4% of the walking sample. Logistic regression models estimated the influence of factors on crawling vs. not crawling or walking vs. not walking. Two models were tested: 1) included sex, age, SES, HAZ and WHZ; 2) added anemia-iron status category to Model 1. HAZ improved the odds of crawling by 30%, but was not significant in either model. Model 2 fit the walking sample data best (P < 0.0001); an increase in HAZ doubled the odds of walking and nonanemic, noniron deficient children were 66% more likely to walk than those with anemia and/or iron deficiency. In this sample of poorly nourished infants, growth and anemia-iron status are significant predictors of walking, but not crawling. PMID- 15795441 TI - Food insecurity and hunger are prevalent among HIV-positive individuals in British Columbia, Canada. AB - Hunger and food insecurity are important factors that may affect an individual's nutritional state and should therefore be assessed in nutrition surveillance activities. The objective of this study was to determine the level of food insecurity and hunger among HIV-positive persons accessing antiretroviral therapy in British Columbia. A cross-sectional study was performed in the BC HIV/AIDS drug treatment program, a province-wide source of free-of-charge antiretroviral medications. In 1998-1999, participants completed a questionnaire focusing on personal information, health, and clinical status. Food and hunger issues were evaluated with the Radimer/Cornell questionnaire. Overall, 1213 responding men and women were classified as food secure (52%), food insecure without hunger (27%), or food insecure with hunger (21%). In both categories of food insecurity, individuals were significantly more likely to be women, aboriginals, living with children, and to have less education, a history of recreational injection drug and/or alcohol abuse, and an unstable housing situation (P < 0.05). In logistic multivariate modeling, income < or = Can$10,000 [adjusted odds ratio (AOR) 3.78, 95% CI (2.53-5.65)], shared household with children [AOR 3.68, 95% CI (1.98 6.84)] and unemployment [AOR 3.15, 95% CI (1.94-5.13)] were the strongest predictors of hunger. In HIV-positive individuals, the occurrence of food insecurity was nearly 5 times higher than in the general Canadian population. The results should stimulate further research to identify to what extent hunger associated factors are reversible with interventions built on nutritional and/or social strategies. PMID- 15795442 TI - Ad libitum choline intake in healthy individuals meets or exceeds the proposed adequate intake level. AB - Choline is an essential nutrient for humans that is used to synthesize membrane phospholipids and the neurotransmitter acetylcholine. Betaine, a metabolite of choline, functions as a methyl-group donor in the conversion of homocysteine to methionine, and is important for renal function. Accurate analysis of choline intake was previously not possible because the choline content of most foods was not known. Using new and recently published data on the concentrations of choline in common foods, we measured the choline content of diets consumed ad libitum by healthy adult volunteers housed in a clinical research center and compared these with estimates of choline intake derived from 3-d food records kept by subjects immediately before study enrollment. Mean choline intake in this subject population met or slightly exceeded the current Adequate Intake (AI) of 7 mg/(kg . d) set by the Institute of Medicine. Men and women consumed similar amounts of choline per day (8.4 and. 6.7 mg/kg, respectively; P = 0.11). Choline intakes estimated from the 3-d food records were significantly lower than this (when expressed as mg/kg, or as total mg, but not when normalized to energy intake), suggesting underreporting of food intake. Intake of betaine, which may spare choline utilization as a methyl-group donor, was 5.3 mg/(kg . d) in men and 4.7 mg/(kg . d) in women. Intake of folate, vitamin B-12, and methionine + cysteine, were similar and sufficient in all subjects. The current recommended AI for choline seems to be a good approximation of the actual intake of this nutrient. PMID- 15795443 TI - Deficiencies in selenium and/or vitamin E lower the resistance of mice to Heligmosomoides polygyrus infections. AB - Previous studies have shown that deficiencies in selenium (Se) and/or vitamin E (VE) can exacerbate the infectivity and pathogenesis of coxsackievirus B3 and influenza. Both Se and VE play a role in immune function and antioxidant defense. To determine whether these deficiencies would affect the normal course of infection with a metazoan parasite, mice were made deficient in Se and/or VE and inoculated with the gastrointestinal nematode parasite Heligmosomoides polygyrus. Both primary and secondary infections were assessed. Although the course of a primary infection with H. polygyrus was unaffected by diet, diets deficient in Se, VE, and both Se and VE (Se/VE double-deficiency) all caused delayed adult worm expulsion and increased fecundity during a secondary infection; suggesting an impaired intestinal response. H. polygyrus-induced IL-4 levels were diet independent; but Se/VE double-deficiency blocked the H. polygyrus-induced IL-4 receptor-associated decrease in sodium-dependent glucose absorption in the jejunum that contributes to worm expulsion. In contrast, Se/VE double-deficiency had no effect on the infection-induced, IL-4R-associated increase in epithelial cell permeability that accompanies the infection. These results suggest that both Se and VE are required for specific IL-4-related changes in intestinal physiology that promote host protection against H. polygyrus. PMID- 15795444 TI - Dietary fructooligosaccharides increase intestinal permeability in rats. AB - We showed previously that fructooligosaccharides (FOS) decrease the resistance to salmonella infection in rats. However, the mechanism responsible for this effect is unclear. Therefore, we examined whether dietary FOS affects intestinal permeability before and after infection with Salmonella enterica serovar Enteritidis. Male Wistar rats were fed restricted quantities of a purified diet that mimicked the composition of a Western human diet. The diet was supplemented with 60 g/kg cellulose (control) or 60 g/kg FOS and with 4 mmol/kg of the intestinal permeability marker chromium EDTA (CrEDTA) (n = 8 or 10). After an adaptation period of 2 wk, rats were orally infected with 10(8) colony-forming units (cfu) of S. enteritidis. Mucin concentrations in intestinal contents and mucosa were measured fluorimetrically, as markers of mucosal irritation. Intestinal permeability was determined by measuring urinary CrEDTA excretion. Translocation of salmonella was quantified by analysis of urinary nitric oxide metabolites with time. Before infection, FOS increased mucosal lactobacilli and enterobacteria in cecum and colon, but not in the ileum. However, FOS increased cytotoxicity of fecal water and intestinal permeability. Moreover, FOS increased fecal mucin excretion and mucin concentrations in cecal and colonic contents, and in cecal mucosa before infection. After infection, mucin excretion and intestinal permeability in the FOS groups increased even further in contrast to the control group. In addition, FOS increased translocation of salmonella to extraintestinal sites. Thus, FOS impairs the intestinal barrier in rats, as indicated by higher intestinal permeability. Whether these results can be extrapolated to humans requires further investigation. PMID- 15795445 TI - Dietary patterns using the Food Guide Pyramid groups are associated with sociodemographic and lifestyle factors: the multiethnic cohort study. AB - Dietary patterns have been used to identify typical combinations of foods that may be associated with disease risks. We defined dietary patterns among 195,298 participants of the Multiethnic Cohort Study in Hawaii and Los Angeles in 1993 1996. Intakes of Food Guide Pyramid groups were calculated from a quantitative FFQ for subjects of 5 ethnic groups (African Americans, Hawaiians, Japanese Americans, Latinos, and whites). Three distinct dietary patterns, "Fat and Meat," "Vegetables," and "Fruit and Milk," were identified by exploratory factor analysis with a varimax rotation and validated by confirmatory factor analysis. Similar factor loadings were found for each of 10 ethnic-gender groups in stratified analyses. The odds ratios (OR) for being above the median scores for each factor were calculated. Age, gender, and ethnicity had relatively strong associations with dietary patterns whereas education showed only weak associations. BMI > or = 30 was strongly positively associated with the Fat and Meat pattern (OR = 2.14, 95% CI: 2.08-2.20, vs. BMI < 25). Current smokers showed a positive association with the Fat and Meat pattern (OR = 1.67, CI: 1.62-1.72, vs. nonsmokers) and inverse associations with the Vegetables (OR = 0.66, CI: 0.64 0.68) and Fruit and Milk patterns (OR = 0.53, CI: 0.52-0.55). Physical activity was positively associated with the Vegetables and Fruit and Milk patterns but not with the Fat and Meat pattern. These findings support the hypothesis that dietary patterns are influenced by interrelated sociocultural, demographic, and other lifestyle factors and may be useful in investigations of diet-disease relations. PMID- 15795446 TI - Mother-infant interactions and infant development are altered by maternal iron deficiency anemia. AB - The aim of this study was to determine whether iron deficiency anemia (IDA) in young South African mothers alters mother-infant interactions and the infant's development. The study was a prospective, randomized, controlled intervention trial with 3 groups of mothers: nonanemic controls and anemic mothers administered either placebo (25 mg ascorbic acid and 10 microg folate) or daily iron treatment (125 mg FeSO(4) plus ascorbate and folate). Mothers of full-term, normal birth weight infants (n = 81) were followed from 10 wk to 9 mo postpartum. Maternal iron status, socioeconomic level, mother-infant interaction [Parent/Caregiver Involvement Scale (PCIS scale)], and infant development (Griffiths scale) were assessed. At baseline, anemic mothers tended (P < 0.10) to be less responsive to, and more controlling of, their infants. Infants of anemic mothers were developmentally delayed at 10 wk in hand-eye movement and overall quotient. Despite normalization of maternal iron status with supplementation in some mothers, the developmental delays were not diminished at 9 mo. At 9 mo, anemic mothers were significantly more "negative" towards their babies, engaged less in goal setting, and were less "responsive" than control mothers. In contrast, the behavior of anemic mothers given iron treatment toward their children was similar to that of the control mothers on all 11 scales of the PCIS. In conclusion, IDA altered mother-child interactions at both 10 wk and 9 mo postpartum. Additionally, infants whose mothers were anemic in the early postpartum scored worse on developmental tests at 10 wk and 9 mo of age. PMID- 15795447 TI - Dietary intakes vary with age among Eskimo adults of Northwest Alaska in the GOCADAN study, 2000-2003. AB - Dietary factors influence the development of cardiovascular disease (CVD). The diet of Alaskan Eskimos differs from that of other populations. We surveyed Eskimo adults in Northwest Alaska to document their usual dietary intakes, differences based on gender and age, and sources of selected nutrients, and to generate appropriate dietary advice to reduce CVD. Interviewers surveyed 850 men and women 17-92 y old, using a quantitative food-frequency instrument. We observed many significant (chi(2) analysis P < 0.05) differences in nutrient intakes among 3 age-groups. Energy intake from carbohydrate was negatively related to participant age-group (P < or = 0.01). Energy intake from all fats (P < 0.001) and polyunsaturated fat (P < or = 0.01) was positively related to age group among both men and women in contrast to other studies in which age differences were either not observed or decreased with age. Native foods were major sources of monounsaturated and polyunsaturated fats, including 56% of (n-3) fatty acids primarily from seal oil and salmon. However, Native foods contributed significantly less to the diets of young adults than to those of elders, especially among women. Store-bought foods were the main sources of energy, carbohydrate, fat, saturated fat, and fiber for all adults. Based on their nutrient density and potential to inhibit CVD, continued consumption of traditional foods is recommended. Variations in intake by age may portend changing eating patterns that will influence CVD as participants age. These data will contribute to understanding dietary risk factors for cardiovascular disease in this population. PMID- 15795448 TI - Dietary iron positively influences bone mineral density in postmenopausal women on hormone replacement therapy. AB - The associations of dietary intakes of iron and calcium on change in bone mineral density (BMD) were examined over 1 y in healthy nonsmoking postmenopausal women (mean age 55.6 +/- 4.6 y) stratified by hormone replacement therapy (HRT) use (HRT, n = 116; no HRT, n = 112). BMD was measured at lumbar spine L(2)-L(4), trochanter, femur neck, Ward's triangle, and total body using dual-energy X-ray absorptiometry at baseline and 1 y. Mean nutrient intakes were assessed using 8-d diet records. All women received 800 mg/d of supplemental elemental calcium. Regression analyses examined the effects of iron and calcium intakes on BMD change adjusting for years past menopause, baseline BMD, weight change, exercise, and energy intake. The interaction of iron with calcium on BMD change was assessed using tertiles of iron and calcium intake and estimated marginal mean change in BMD. Iron was associated (P < or = 0.05) with greater positive BMD change at the trochanter and Ward's triangle in women using HRT. Calcium was associated (P < or = 0.05) with BMD change at the trochanter and femur neck for women not using HRT. In women using HRT in the lowest tertile of calcium intake, change in femur neck BMD increased linearly as iron intake increased. In women not using HRT, BMD increased in the women in the highest tertile of calcium intake. We conclude that HRT use appears to influence the associations of iron and calcium on change in BMD. PMID- 15795450 TI - Introduction. PMID- 15795449 TI - Citrus limonoids induce apoptosis in human neuroblastoma cells and have radical scavenging activity. AB - Citrus limonoid glucosides, a family of fruit bioactive compounds, were postulated to have free radical-scavenging and apoptosis-inducing properties against certain types of cancers. Four highly purified limonoid glucosides, limoin 17beta D-glucopypranoside (LG), obacunone 17beta D-glucopyranoside (OG), nomilinic acid 17beta D-glucopyranoside (NAG), and deacetylnomilinic acid 17beta D-glucopyranoside (DNAG) were tested for superoxide radical (O(2)(-))-quenching activity and cytotoxic action against undifferentiated human SH-SY5Y neuroblastoma cells in culture. All 4 scavenged O(2)(-) as measured by inhibition of pyrogallol decomposition in a spectrophotometric assay. Quenching by NAG in particular emulated an equivalent concentration of vitamin C. When added to the medium of SH-SY5Y cells in culture, micromolar amounts of LG and OG, compared with untreated controls, caused a cessation of cell growth and rapid cell death (P < 0.001); NAG and DNAG were better tolerated, but nonetheless toxic as well. Cytotoxicity was related to a concentration- and time-dependent increase in caspase 3/7 activity, suggesting that limonoid glucosides were capable of inducing apoptosis. Arrested cell growth and the induction of apoptosis were confirmed by flow cytometry and DNA fragmentation analysis. Importantly, caspase induction at 12 h correlated with cell survival at 24 h (P = 0.046), suggesting that apoptosis was the primary cause of cell death. We conclude that citrus limonoid glucosides are toxic to SH-SY5Y cancer cells. Cytotoxicity is exerted through apoptosis by an as yet unknown mechanism of induction. Individual limonoid glucosides differ in efficacy as anticancer agents, and this difference may reside in structural variations in the A ring of the limonoid molecule. PMID- 15795451 TI - Methodological challenges in performing targeting: assessing dietary risk for WIC participation and education. AB - This paper summarizes recent evaluations of indicators of dietary risk used to determine eligibility and nutrition education and counseling within the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). The concept of dietary risk relates to an inadequate dietary pattern or a failure to adhere to the food guide pyramid and, within WIC, is typically assessed based on an individual's intake from a 24-h recall or FFQ. Available evidence suggests, however, that even with a high-quality technique, substantial error exists in these data at the level of the individual, making the likelihood of misclassification high. Such data, with appropriate statistical procedures, can provide valid information at the group or the population level, and this is a future area for indicator development and incorporation into the management of WIC and other nutrition education programs addressing dietary risk. PMID- 15795452 TI - Meanings of targeting from program workers. AB - Program workers have a critical role in targeting in nutrition programs, but little has been studied about the concept and the operation of targeting from their perspectives. This paper introduces and compares meanings of targeting from the perspectives of program workers in food assistance programs in the United States. There exist some variations in perceptions of targeting and actual operational procedures adopted by program workers at the local and federal levels. Regardless, program workers' perspectives on targeting suggest a new framework of targeting, which consists of key components (program access, service, and evaluation) and issues in need of careful attention in targeting nutrition programs. The success of targeting depends on how well these components and related issues can be translated into targeting policy, rules, and procedures. With increased devolution to the state and local levels in the United States, it is even more critical for federal policy makers and administrators to understand how frontline program workers perceive and implement targeting. The new framework based on the perspectives of program workers may contribute to developing a conceptual definition as well as practical guidance for targeting that can be used to formulate better ways of targeting in nutrition programs. PMID- 15795453 TI - Interactions with the recipient community in targeted food and nutrition programs. AB - This article explores the relation between targeted nutrition and food assistance programs and the recipient communities. The author begins by suggesting that the renewed current emphasis on targeting has resulted from the high cost of universal entitlements in poor countries and from the need to increase the food security and resilience of the poor in these countries. They stress the fundamental human right of communities to have involvement in all steps of program targeting as elemental to good democratic governance. Next, the article reviews the issues surrounding different levels of community involvement in the components of program targeting, namely needs assessment, definition and implementation of eligibility criteria, delivery of services and benefits, and monitoring and evaluation of results. Four types of targeting eligibility are described: self-targeting, means tested, categorical, and community based. It is noted that the type and the level of engagement of programs with communities depends on whose concept of community welfare is being pursued (the development summit's or the community's), and on how broadly nutrition improvement is defined. Examples are used to illustrate the potentially severe consequences of ignoring community governance and social structures in the targeting of programs. A framework is then presented with important variables to consider when planning a targeted program: state governance contexts, the nature of the local community's institutions of governance, the ratio of the need to the available benefit, and the stability of the context. Finally, the article presents recommendations for filling in the large research gaps on this topic. PMID- 15795454 TI - The science and politics of targeting: who gets what, when, and how. AB - Targeting has been defined as a method by which goods, services, or other programmatic actions are delivered to groups or individuals who have specific characteristics. Similarly, politics refers to the social processes that determine who gets what, when, and how. This similarity suggests there may be utility in undertaking an explicit analysis of how the science and the politics of targeting relate to one another. Accordingly, this article outlines a technical/scientific framework regarding targeting and, using one element of this framework [sensitivity and specificity analysis], demonstrates that recent shifts in politics and policy in the United States can be more clearly understood when viewed from this angle. Then, the article outlines a basic framework on politics and uses it to demonstrate that one of the most fundamental features of our training, practices, and outlook (specialization) holds obvious benefits for advancing research but has three undesirable effects in the realm of policy: 1) it directly or indirectly encourages narrow, partial solutions to complex problems, some of which may have unintended effects on health or well-being for the very people we seek to help; 2) it creates a cumulative demand for research dollars, specialized interventions, and policy attention that far exceeds available resources and capabilities; and 3) it often prevents us from developing and advancing the common agendas and mutual support required to be effective in political and policy realms. The article concludes that more explicit analysis along these lines could strengthen our effectiveness in the policy realm. PMID- 15795455 TI - How to modify the food environment. PMID- 15795456 TI - Food choices and diet costs: an economic analysis. AB - Obesity in the United States is a socioeconomic issue. It is related to limited social and economic resources and may be linked to disparities in access to healthy foods. Added sugars and added fats are far more affordable than are the recommended "healthful" diets based on lean meats, whole grains, and fresh vegetables and fruit. There is an inverse relationship between energy density of foods (kJ/g) and energy cost ($/MJ), such that energy-dense grains, fats, and sweets represent the lowest-cost dietary options to the consumer. Good taste, high convenience, and the low cost of energy-dense foods, in conjunction with large portions and low satiating power, may be the principal reasons for overeating and weight gain. Financial disparities in access to healthier diets may help explain why the highest rates of obesity and diabetes are found among minorities and the working poor. If so, then encouraging low-income households to consume more costly foods is not an effective strategy for public health. What is needed is a comprehensive policy approach that takes behavioral nutrition and the economics of food choice into account. PMID- 15795458 TI - Public health strategies for dietary change: schools and workplaces. AB - Environmental intervention represents an effective strategy to promote the purchase and consumption of healthful foods in community-based settings. Three such studies promoted the consumption of healthier snacks and lower-fat foods at worksites and at schools. The first study examined lower prices and point-of purchase promotion on sales of lower-fat snacks in vending machines in 12 worksites and 12 secondary schools. Price reductions of 10%, 25% and 50% on lower fat snacks resulted in an increase in sales of 9%, 39% and 93%, respectively, as compared to usual-price conditions. The second study examined the impact of a 50% price reduction on the consumption of fresh fruit and baby carrots in 2 secondary school cafeterias. In the third study, 20 secondary schools were randomly assigned to either an environmental intervention or a control group for a 2-y period. The intervention increased the availability of lower-fat foods and implemented student-based promotions. School-based environmental interventions to increase availability and promotion of lower-fat foods and healthier snacks can increase purchase of these foods among adolescents. PMID- 15795457 TI - Portion sizes and the obesity epidemic. AB - The rise in obesity rates over the past 30 y has been paralleled by increases in the portion size of many foods and the prevalence of eating away from home. Foods of particular concern are those that have a high energy density (kJ/g). Many well controlled, laboratory-based studies have found that large portions of energy dense foods can lead to excess energy intakes. This influence of large portions on energy intake has been supported by data collected in naturalistic settings. Further research is needed to explore strategies that can be used to moderate the effects of portion size on food consumption. One promising strategy is to reduce the energy density of foods, while maintaining food weight or volume, so that consumers can eat satisfying portions while reducing their energy intakes. There is a need for effective educational messages that not only emphasize limiting the consumption of foods high in energy density, but also encourage the consumption of those with a low energy density, such as fruits and vegetables. The delivery of consistent messages will require more cooperation among the food and restaurant industries, policy makers, and scientists. Effective strategies will also require consumers to understand and accept the importance of eating reasonable portions for better health. PMID- 15795459 TI - The global face of nutrition: what can governments and industry do? AB - Investment in agricultural research and major developments in the technology of food production have led to dramatic increases in food availability, not only in industrialized nations but also across the globe. In addition, significant gains in improving nutritional status have been made over the past fifty years. These changes are the result of 1) the demographic transition, 2) the epidemiological transition, and 3) the nutrition transition. Countries throughout the world are at various stages of the nutrition transition, as traditional diets give way to eating patterns characterized by more animal products, more added sugar, and more added fat. The world health organizations now face a different dilemma. Increasingly, countries are documenting that food insecurity and undernutrition exist side by side with problems of overnutrition and chronic diseases. Investment in applied nutrition research is desperately needed to identify and implement effective approaches for promoting healthy lifestyles. PMID- 15795460 TI - Bioethics in scientific research: conflicts between subject's equitable access to participate in research and current regulations. PMID- 15795461 TI - Informed consent in the context of communities. AB - This article discusses the history of institutional review boards and the change from a focus on protection of subjects from harm from research to a concern for equitable access to research protocols and potential benefits from participating in a study. The concern about an overemphasis on potential benefits to the detriment of individual subjects is discussed. Finally, the distinction between protection of individuals as it is separate from communities, issues that are raised about obtaining informed consent in communities, as well as the need for trust among researchers, individuals, and communities is examined. PMID- 15795462 TI - Looking into the institutional review board: observations from both sides of the table. AB - Institutional review board (IRB) reviews offer the benefit of perspective afforded by the board's distance from the research and the research subjects. At the same time, distance from research subjects that is geographic, socioeconomic, cognitive, linguistic, and cultural can undermine the positive role of perspective. In addition, distance between IRB and investigators, largely a result of attitudes and communication, can prolong the review process and can obscure its message. The tension that often characterizes IRB-investigator relationships is due, in part, to variability in the application of federal regulations by IRBs across institutions and, on the part of investigators, inexperience, communication problems, and difficulties in anticipating the needs of their subjects. Contributing to the variability are the demographics and the culture of the IRB, attitudes that influence IRB-investigator relationships, and the adequacy of support from the institution. The effects of these factors on review decisions and on the performance of the human subjects protection system are largely unstudied. The movement for IRB accreditation is causing institutions to examine their overall research protection system and promises a more collaborative approach, where IRB and investigators accept their common charge to meet the needs of subjects and to improve the quality of research. PMID- 15795463 TI - Conducting research in developing countries: experiences of the informed consent process from community studies in Peru. AB - This article discusses the process for obtaining genuine informed consent for the participation of human subjects in research in developing countries. We discuss the consent process in the light of recently published guidelines, the experience of nutrition and health research projects, and the ethics review process of the Instituto de Investigacion Nutricional with peri-urban and rural populations in Peru. We discuss the cultural context in relation to (i) who should be involved in the decision for participation, especially for research in children and in community settings; (ii) when to use written or verbal consent; (iii) the format and presentation of the consent form to ensure understanding by the target population; and (iv) the process of how and by whom information is given and consent is obtained. Common concerns of participants with regard to their involvement in research studies are presented, as well as aspects that participants find difficult to understand. Some specific concerns of conducting research with Indigenous Peoples are discussed. We recommend future research to further understand and implement informed consent processes to assure genuine and voluntary consent in different developing country contexts. PMID- 15795464 TI - Balancing access to participation in research and protection from risks: applying the principle of justice. AB - The problem for Institutional Review Boards (IRBs) of balancing access to participation in research with protection of research subjects has always been a difficult one. IRBs, charged with applying the "Common Rule," as well as the Belmont Principles, in their review of clinical research, are given little guidance on approaching this problem. This article argues that the third Belmont Principle, the Justice Principle, may provide a useful framework for considering this balance. The changing research environment is discussed in an historical context, and the Justice Principle is considered both in the context of individual rights, as well as the potential benefit to classes of people. The author further suggests that application of the Justice Principle be driven by findings derived from an analysis of the first 2 principles. This feedback model will enable a more formal application of the Justice Principle and less ambiguous, more transparent, decisions regarding the equitable selection of subjects. The author calls for more systematic attention to the Justice Principle by IRBs, and proposes a model that includes incorporating the deliberation of the other Belmont Principles into the Justice Principle. PMID- 15795465 TI - Women's voices, women's choices: the challenge of nutrition and HIV/AIDS. AB - The Society for International Nutrition Research sponsored a Symposium titled "Women's Voices, Women's Choices: The Challenge of Nutrition and HIV/AIDS in Asia and Africa" at Experimental Biology 2004 to highlight the challenges facing HIV positive women living in resource-poor settings of Asia and Africa, when it comes to the everyday decisions they are forced to make about their own health and nutrition, and the health and the nutrition of their children. This introductory paper summarizes the rationale for this session, including a summary of the evidence for women's increased vulnerability to HIV, the nutritional impacts of HIV infection, and the special infant feeding and nutritional concerns facing HIV positive pregnant and lactating mothers in Africa and Asia. The issue of nutrition and HIV/AIDS is addressed here from an intergenerational perspective, using new data from qualitative research, clinical trials, and behavioral interventions in India, Malawi, South Africa, Tanzania, and Zimbabwe, to illustrate important concerns, using study participants' own words to convey key messages. The focus is on women, because they are shouldering much of the burden of HIV infection in terms of their numbers and in their responsibilities for providing food and care for orphans and HIV-affected family members. Infant feeding choices are also considered in this review, because of the vast implications that not breast-feeding at all and stopping breast-feeding early have on the nutritional well-being of HIV-exposed children, as well as the positive contribution of breast-feeding to child nutrition and survival worldwide. PMID- 15795466 TI - Studies of vitamins and minerals and HIV transmission and disease progression. AB - HIV-1 infection is having a devastating impact on people in developing countries. Poor nutrition and HIV-related adverse health outcomes contribute to a vicious cycle that may be slowed down by using nutritional interventions, including vitamins and minerals. Among children, periodic supplementation with vitamin A starting at 6 mo of age has been shown to be beneficial in reducing mortality and morbidity among both HIV-infected and uninfected children. Limited data exist on the role of other nutrient supplements among children. Among HIV-infected adults, the safety and the efficacy of vitamin A supplements need further study, although adequate dietary intake of this essential nutrient is recommended. Multivitamin supplements were efficacious in reducing adverse pregnancy outcomes and early childhood infections, and is currently provided to pregnant HIV-infected pregnant women in many programs. The efficacy of such supplements among HIV-negative pregnant women needs further study. Daily multivitamin supplements were found to reduce HIV disease progression among men and women in several observational studies and randomized trials, and to provide an important low-cost intervention that could be provided to adults in early stages of HIV disease to prolong the time before antiretroviral therapy is recommended. Next, research priorities include examining the roles of minerals, including selenium, in HIV infection, as well as determining the safety and the efficacy of micronutrient supplements among individuals who are advanced in their disease and who are receiving antiretroviral therapy. PMID- 15795467 TI - Perceptions of the role of maternal nutrition in HIV-positive breast-feeding women in Malawi. AB - A neglected issue in the literature on maternal nutrition and HIV is how HIV positive women perceive their own bodies, health, and well-being, particularly in light of their infection, and whether these perceptions influence their infant feeding practices and their perceived ability to breast-feed exclusively through 6 mo. We conducted formative research to better understand breast-feeding practices and perceptions, and to inform the Breastfeeding, Antiretroviral, and Nutrition (BAN) Study, a clinical trial to evaluate antiretroviral and nutrition interventions to reduce mother-to-child transmission of HIV during breast-feeding in Lilongwe, Malawi. Twenty-two HIV-positive women living in semi-rural areas on the periphery of Lilongwe participated in in-depth interviews. In an adaptation of the body-silhouette methodology, nine culturally appropriate body silhouettes, representing a continuum of very thin to very large shapes, were used to elicit women's views on their present, previous-year, and preferred body shapes, and on the shape they perceived as healthy. The narrative scenario method was also used to explore women's views on 2 fictional women infected with HIV and their ability to exclusively breast-feed. Women perceived larger body shapes as healthy, because fatness is considered a sign of good health and absence of disease, and many recognized the role of nutrition in achieving a preferred or healthy body shape. Several women believed their nutritional status (body size) was declining because of their illness. Women were concerned that breast-feeding may increase the progression of HIV, suggesting that international guidelines to promote appropriate infant feeding practices for infants whose mothers are infected with HIV should focus on the mother's health and well-being, as well as the infant's. PMID- 15795468 TI - An education and counseling program for preventing breast-feeding-associated HIV transmission in Zimbabwe: design and impact on maternal knowledge and behavior. AB - International guidance on HIV and infant feeding has evolved over the last decade. In response to these changes, we designed, implemented, and evaluated an education and counseling program for new mothers in Harare, Zimbabwe. The program was implemented within the ZVITAMBO trial, in which 14,110 mother-baby pairs were enrolled within 96 h of delivery and were followed at 6 wk, 3 mo, and 3-mo intervals. Mothers were tested for HIV at delivery but were not required to learn their test results. Infant feeding patterns were determined using data provided up to 3 mo. Formative research was undertaken to guide the design of the program that included group education, individual counseling, videos, and brochures. The program was introduced over a 2-mo period: 11,362, 1311, and 1437 women were enrolled into the trial before, during, and after this period. Exclusive breast feeding was recommended for mothers of unknown or negative HIV status, and for HIV-positive mothers who chose to breast-feed. A questionnaire assessing HIV knowledge and exposure to the program was administered to 1996 mothers enrolling after the program was initiated. HIV knowledge improved with increasing exposure to the program. Mothers who enrolled when the program was being fully implemented were 70% more likely to learn their HIV status early (<3 mo) and 8.4 times more likely to exclusively breast-feed than mothers who enrolled before the program began. Formative research aided in the design of a culturally sensitive intervention. The intervention increased relevant knowledge and improved feeding practices among women who primarily did not know their HIV status. PMID- 15795469 TI - Infant feeding dilemmas created by HIV: South African experiences. AB - Breast-feeding is a route of HIV transmission from an HIV infected mother to her infant. However, breast-feeding is an important pillar of child survival and part of a mother's womanhood. This paper highlights the dilemma created by the risks and the benefits of breast-feeding and will discuss the implementation in South Africa, of the Safer Breastfeeding Programme, to reduce some of the known risk factors associated with HIV transmission. Operations research was carried out, including infant HIV testing at 6 wk, and 9, 12, and 15 mo. Among 188 breast-fed infants enrolled in the program who were HIV negative at 6 wk, 4 became infected by 9 mo of age (2.6%). Infants enrolled in the Safer Breastfeeding Programme had less breast pathology than is usually reported for HIV infected women. Limited success was attained in promoting heat treatment of expressed breast milk; however, it does appear to be a feasible option after 6 mo of age, and it is believed that mothers would practice it more widely if sufficient promotion and support was given to it. Mothers found it difficult to stop breast-feeding earlier than the norm, and it therefore is important that mothers considering early cessation of breast-feeding be given sufficient preparation and support. PMID- 15795471 TI - TGF-beta signaling pathway: its role in gastrointestinal pathophysiology and modulation of ulcer healing. AB - Gastrointestinal ulcer healing is a complex process, involving cell migration, proliferation, angiogenesis and extracellular matrix deposition, all ultimately leading to reconstruction of tissue architecture within the ulcer scar. These processes are controlled by growth factors, cytokines and hormones. Transforming growth factor-beta (TGF-beta), one of the multifunctional peptide growth factors, has been reported to positively regulate gastrointestinal ulcer healing. Although TGF-beta inhibits cell proliferation in a variety of cells, it induces cell migration, angiogenesis, and enhances extracellular matrix production necessary for gastrointestinal ulcer healing. TGF-beta exerts its action by binding to its transmembrane serine/threonine kinase receptors, which in turn triggers activation of various intracellular signaling pathways. Smads are intermediate effector proteins that play key roles in biological activities of TGF-beta by transmitting the signals from the cell surface directly into the nucleus and initiating transcription. New insight into the mechanisms underlying TGF-beta Smad modulation of gastrointestinal ulcer healing will likely enhance our understanding of the mechanisms controlling the healing processes of gastrointestinal ulcers. PMID- 15795470 TI - Making the choice: the translation of global HIV and infant feeding policy to local practice among mothers in Pune, India. AB - In 2003, India had over 5.1 million infected individuals living with HIV/AIDS. The percentage of all HIV cases attributed to perinatal transmission has been increasing steadily from 0.33% of total cases in 1999 to 2.80% in 2004. Recent statistics indicate that over 130,000 infants have been infected through this route. Despite recent advances in reducing in utero and interpartum transmission with the use of antiretrovirals, there is a critical need to make infant feeding safer. Current UNAIDS/WHO/UNICEF recommendations stress avoidance of all breast feeding if replacement feeding fulfills the key requirements of being affordable, feasible, acceptable, sustainable, and safe. In this paper, we examine how the UNAIDS/WHO/UNICEF recommendations have been actualized within the context of an urban government hospital in India. The documented patterns of infant feeding by HIV-positive mothers in Pune, India, from 2000 to 2004, highlight the complexities of making an informed and healthy choice under suboptimal conditions. The data indicate that interpersonal variations in the key requirements greatly influence the optimal practice to minimize mortality risks. Moreover, local information on health outcomes is crucial to tailoring policy recommendations to save lives. We propose the development of a decision-making algorithm that includes factors affecting mother-to-infant transmission, including site-specific data on health risks to the mother and the child. Such an algorithm would allow identification of the healthiest feeding choice and would minimize the pitfalls of promoting homogeneous practices lacking site-specific evidence-based evaluation. PMID- 15795472 TI - Role of p53 tumor suppressor in ageing: regulation of transient cell cycle arrest and terminal senescence. AB - In this study we investigated the function of p53 as a regulator of cell cycle progression in cycling and senescent cells. Using the conditional temperature sensitive (ts) mutant we could prevent the detrimental effect of constitutive expression of high levels of wt p53 protein. High levels of wt p53 inhibited cell proliferation by blocking the cells to progress from G1 to S phase of the cell cycle. Flow cytometric analysis revelaed a maintenance of G1 cell population for a longer time depending on the prolonged expression of wt p53 protein. The p53 mediated inhibition of cell proliferation and of the cycle was reversible. However, a spontaneous increase of wt p53 occurring in ageing normal human MRC-5 fibroblasts was associated with irreversible reduction of proliferative potential. The accumulation of G1 cells was detected by flow cytometry. By the measurement of DNA content it is not possible to discriminate between cells arrested in G1 and G0 phase, therefore, the expression of G1 markers was determined. Analysis of the expression of distinct cell cycle regulators revealed that quiescent MRC-5 cells were in G0 phase. Our results indicate that cell cycle arrest occurring in senescent cells is associated with the G0 transition. PMID- 15795474 TI - Estradiol modulates the synapsins phosphorylation by various protein kinases in the rat brain under in vitro and in vivo conditions. AB - Synapsins are the neuronal phosphoproteins which play very important role in processes of synaptic neurotransmission. They are physiological substrates for Ser/Thr protein kinases. The reversible phosphorylation of synapsins may be modified by several compounds including steroid hormones. The aim of our study was to investigate, if the one of neuroactive steroid--17beta-estradiol--could modulate the phosphorylation of synapsins by PKA, CaM-PK and PKC in rat brain and what type of mechanism of their action is possible. The activity of kinases was evaluated as phosphorylation of synapsin in cerebral cortex and hippocampus in vivo and in vitro conditions. We conclude that 17E2 has inhibitory effect on synapsins phosphorylation by all tested kinases in vitro and in vivo conditions. The lack of nuclei in synaptosomal membrane fraction and short time of hormone exposure can be evidence of direct, non-genomic mechanism of estradiol action. PMID- 15795473 TI - Expression of alpha-synuclein in different brain parts of adult and aged rats. AB - The synucleins are a family of presynaptic proteins that are abundant in neurons and include alpha-, beta, and gamma-synuclein. Alpha-synuclein (ASN) is involved in several neurodegenerative age-related disorders but its relevance in physiological aging is unknown. In the present study we investigated the expression of ASN mRNA and protein in the different brain parts of the adult (4 month-old) and aged (24-month-old) rats by using RT-PCR technique and Western blot, respectively. Our results indicated that mRNA expression and immunoreactivity of ASN is similar in brain cortex, hippocampus and striatum but markedly lower in cerebellum comparing to the other brain parts. Aging lowers ASN mRNA expression in striatum and cerebellum by about 40%. The immunoreactivity of ASN in synaptic plasma membranes (SPM) from aged brain cortex, hippocampus and cerebellum is significantly lower comparing to adult by 39%, 24% and 65%, respectively. Beta-synuclein (BSN) was not changed in aged brain comparing to adult. Age-related alteration of ASN may affect the nerve terminals structure and function. PMID- 15795475 TI - Effect of combined treatment with imipramine and metyrapone on the immobility time, the activity of hypothalamo-pituitary-adrenocortical axis and immunological parameters in the forced swimming test in the rat. AB - Major depression is frequently associated with the hyperactivity of the hypothalamic-pituitary-adrenocortical axis, and glucocorticoid synthesis inhibitors have been shown to exert antidepressant action. The aim of the present study was to examine the effect of joint administration of metyrapone (50 mg/kg) and imipramine (5 and/or 10 mg/kg) on immobility time, plasma corticosterone concentration, the weight of spleens and thymuses and the proliferative activity of splenocytes in rats subjected to the forced swimming test--an animal model of depression. Metyrapone alone (50 mg/kg) reduced the immobility time of rats in the forced swimming test and decreased plasma corticosterone level, but did not change immunological parameters. Joint administration of metyrapone and imipramine (5 and 10 mg/kg) produced a more pronounced antidepressant-like effect than either of the drugs given alone. The forced swimming procedure significantly increased the proliferative activity of splenocytes, that parameter being reduced only by co-administration of metyrapone and imipramine. Joint administration of metyrapone and imipramine inhibited to a similar extend the corticosterone level as did treatment with metyrapone alone (about twofold); however, the plasma corticosterone level in animals treated with metyrapone and the higher dose of imipramine did not differ from the concentration of this steroid in control, not stressed rats. The obtained results indicate that metyrapone potentiates the antidepressant-like activity of imipramine and exerts a beneficial effect on the stress-induced increase in plasma corticosterone concentration and the proliferative activity of splenocytes. These finding suggest that a combination of metyrapone and an antidepressant drug may be useful for the treatment drug resistant depression and/or depression associated with a high cortisol level. PMID- 15795476 TI - Effect of moderate incremental exercise, performed in fed and fasted state on cardio-respiratory variables and leptin and ghrelin concentrations in young healthy men. AB - BACKGROUND: Although hormonal responses to exercise performed in fed state are well documented, far less in known about the effect of a single exercise bout, performed after overnight fasting, on cardio-respiratory responses and hormones secretion. It has been reported that recently discovered hormones as leptin and ghrelin may affect cardiovascular responses at rest. However, their effect on the cardiovascular responses to exercise is unknown. AIMS: This study was designed to determine the effect of overnight fasting on cardio- respiratory responses during moderate incremental exercise. We have hypothesised that fasting / exercise induced changes in plasma leptin / ghrelin concentrations may influence cardiovascular response. MATERIAL AND METHODS: Eight healthy non-smoking men (means +/- SE.: age 23.0 +/- 0.5 years; body mass 71.9 +/- 1.5 kg; height 179.1 +/- 0.8 cm; BMI 22.42 +/- 0.49 kg x m(-2) with VO2max of 3.71 +/- 0.10 l x min( 1)) volunteered for this study. The subjects performed twice an incremental exercise test, with the increase of power output by 30 W every 3 minutes. Tests were performed in a random order: once in the feed state--cycling until exhaustion and second, about one week later, after overnight fasting--cycling until reaching 150 W. RESULTS: In the present study we have compared the results obtained during incremental exercise performed only up to 150 W (59 +/- 2 % of VO2max) both in fed and fasted state. Heart rate measured during exercise at each power output, performed in fasted state was by about 10 bt x min(-1) (p = 0.02) lower then in fed subjects. Respiratory quotient and plasma lactate concentration in fasted state were also significantly (p<0.001) lower than in the fed state. Pre-exercise plasma leptin and ghrelin concentrations were not significantly different in fed and fasted state. Exercise induced increase in hGH was not accompanied by a significant changes in the studied gut hormones such as ghrelin, leptin, and insulin, except for plasma gastrin concentration, which was significantly (p = 0.008) lower in fasting subjects at the power output of 150 W. Plasma [IL-6] at rest before exercise performed in fasted state was significantly (p = 0.03) elevated in relation to the fed state. This was accompanied by significantly higher (p = 0.047) plasma noradrenaline concentration. Plasma IL-6 concentration at rest in fed subjects was negatively correlated with plasma ghrelin concentration (r = -0.73, p < 0.05) and positively correlated with plasma insulin concentration (r = 0.78, p < 0.05). Significant negative correlation (r = -0.90; p < 0.05) was found between plasma insulin and ghrelin concentration at rest in fed subjects. CONCLUSIONS: We have concluded that plasma leptin and ghrelin concentrations have no significant effect on the fasting-induced attenuation of heart rate during exercise. We have postulated that this effect is caused by increased plasma norepinephrine concentration, leading to the increase in systemic vascular resistance and baroreceptor mediated vagal stimulation. Moreover we believe, that the fasting-induced significant increase in plasma IL-6 concentration at rest, accompanied by higher plasma norepinephrine concentration and lower RQ, belongs to the physiological responses, maintaining energy homeostasis in the fasting state. PMID- 15795477 TI - Early effects of short-term endurance training on hormonal responses to graded exercise. AB - Twelve male, sedentary volunteers (22.0 +/-) were submitted to three weeks of a bicycle ergometer training, consisting of 45 min exercise (at 70% VO2max), 4 times in the first week and 3 times in the next 2 weeks. They performed four incremental exercise tests with the power output increased by 50 W every 3 min until volitional exhaustion: two before training (C1 and C2), and after one (T1) and three (T3) weeks of training. Before and after each load the plasma noradrenaline (NA), adrenaline (A) and blood lactate (LA) concentrations were determined in venous blood samples as well as plasma growth hormone (HGH) and cortisol concentrations before and at the end of exercise. A decrease in NA concentration was found already after 1 week of training at power output of 100 W (p<0.01) and 200 W (p<0.05). Similar decline was maintained after 3 weeks of training. No significant training-induced differences in plasma A concentration were found, however, the thresholds for both catecholamines were significantly shifted towards higher values after 3 weeks of training. One week of training caused a decrease in the pre-exercise (p<0.01), as well as post-exercise (p<0.05) plasma cortisol and HGH concentrations. It was concluded that endurance training induced a decrease in HGH, cortisol and NA concentration already after one week of training. A decline of pre-exercise plasma HGH and cortisol levels with time of experiment may, in part, indicate familiarization to exercise protocol. PMID- 15795478 TI - Exercise training and 3-day head down bed rest deconditioning: exercise thermoregulation. AB - Bed rest (BR) deconditioning causes excessive increase of exercise core body tempera-ture, while aerobic training improves exercise thermoregulation. The study was designed to determine whether 3 days of 6 degrees head-down bed rest (HDBR) affects body temperature and sweating dynamics during exercise and, if so, whether endurance training before HDBR modifies these responses. Twelve healthy men (20.7+/-0.9 yrs, VO2max: 46+/-4 ml x kg(-1) x min(-1) ) underwent HDBR twice: before and after 6 weeks of endurance training. Before and after HDBR, the subjects performed 45 min sitting cycle exercise at the same workload equal to 60% of VO2max determined before training. During exercise the VO2, HR, tympanic (Ttymp) and skin (Tsk) temperatures were recorded; sweating dynamics was assayed from a ventilated capsule on chest. Training increased VO2max by 12.1% (p<0.001). Resting Ttymp increased only after first HDBR (by 0.22 +/- 0.08 degrees C, p<0.05), while exercise equilibrium levels of Ttymp were increased (p<0.05) by 0.21 +/- 0.07 and 0.26 +/- 0.08 degrees C after first and second HDBR, respectively. Exercise mean Tsk tended to be lower after both HDBR periods. Total sweat loss and time-course of sweating responses were similar in all exercise tests. The sweating threshold related to Ttymp was elevated (p<0.05) only after first HDBR. IN CONCLUSION: six-week training regimen prevents HDBR-induced elevation of core temperature (Ttymp) at rest but not during ex-ercise. The post HDBR increases of Ttymp without changes in sweating rate and the tendency for lower Tsk suggest an early (<3d) influence of BR on skin blood flow. PMID- 15795479 TI - Effect of cAMP analogues on glomerular inulin space of isolated rats renal glomeruli. AB - Cyclic AMP has been generally recognised as activator of cAMP-dependent protein kinases. However, there is little evidence about role of cAMP-dependent protein kinase (PKA), in particular izoenzymes PKA-I and PKA-II, in glomeruli contractility. We measured changes of glomerular inulin space (GIS) as a marker of its contractility in the presence of phosphodiesterase resistance cAMP analogues; activators and inhibitors of PKA. Activator of PKA i.e. (Sp) 8-Cl cAMPS (0.1-100 microM) decreased GIS. (Rp) 8-Cl-cAMPS (0.1-100 microM), inhibitor of PKA, was ineffective but shifted concentration-response curve of (Sp) 8-Cl cAMPS to right at 50 microM. Specific A site activation by N6-benzoyl-cAMP decreased GIS with maximum at 0.1 microM. Activation of B site by 8 aminobutyloamino-cAMP (0.1-100 microM) had no effect. However, specific activation of both sites on PKA-I or PKA-II by site-selective analogue pairs e.g. 8-aminobutyloamino-cAMP plus 8-piperidino-cAMP or N6-benzoyl-cAMP plus 8 piperidino-cAMP respectively, significantly increased sensitivity of glomeruli to analogues. Our data suggest that activation of PKA-I or PKA-II might have an important role in the regulation of glomerular contractility. PMID- 15795480 TI - The effects of baclofen on the feeding behaviour and body weight of vagally stimulated rats. AB - It is hypothesised that the GABA(B) receptor agonist baclofen increases or has no effect on food intake, and electrical stimulation of vagal nerves decreases food intake. The aim of this study was to evaluate the effects of baclofen in vagally stimulated rats. MATERIAL AND METHODS: Thirty two Wistar rats were divided into five groups: group A scheduled for microchip implantation for vagal stimulation, group B for sham operation, group C for microchip implantation and baclofen medication, group D for baclofen medication only and group E for gastric motility evaluation under influence of baclofen. The following parameters were then evaluated: food intake and body mass, gastric motility, leptin, insulin, and glucose serum levels. RESULTS: In the comparison of groups B and A, daily food intake and body weight gain decreased by 17% (p<0.05) and by 22% (p<0.05), respectively. Baclofen alone (group D) did not significantly change either food intake nor diurnal body weight compared to the controls, but when used in conjunction with the microchip (group C) it did significantly reduce effect of vagal neuromodulation (p<0.05). Furthermore, a significant decrease in leptin and glucose levels was detected in group C: 677 to 165 pg/ml (p<0.05) and 5,93 to 4,88 mmol/l (p<0.05), respectively. The administration of baclofen stimulated significantly gastric motility and elicited irregular motor migrating complex (327+/-200 against control 255+/-52 cmH2O/s). CONCLUSIONS: These results suggest that microchip vagal neuromodulation through increased vagal afferent activity induces an alteration in the feeding behaviour and decreases nocturnal food intake and body weight. These effects were partially attenuated by baclofen. The data suggests that GABA(B) receptors play an important role in the pathomechanism of attenuation of food intake induced by vagal nerve stimulation. PMID- 15795481 TI - Role of afferent nerves and sensory peptides in the mediation of hepatic artery buffer response. AB - Intrahepatic arteries are richly innervated by both adrenergic and sensory vanilloid-sensitive (capsaicin-sensitive) fibers. Stimulation of capsaicin sensitive fibers has been shown to dilate the intrahepatic vessels by both releasing sensory neuropeptides and by modulating the adrenergic tone. However the participation of capsaicin-sensitive fibers in the mediation of the hepatic artery buffer response (HABR) has not been investigated yet. To explore the involvement of sensory innervation and sensory neuropeptides in the HABR, the experiments were performed on capsaicin-denervated Wistar rats. In addition, we used selective CGRP and tachykinin receptor antagonists to test the participation of CGRP, substance P and NK-A in HABR in the rat. In anesthetized rats the hepatic artery blood flow (HABF), microcirculatory hepatic blood flow (HBF) and portal blood flow (PBF) were determined. The HABR was induced by partial occlusion of the portal vein and maintaining the PBF at 10% of its control preocclusive value. In the control HABR the hepatic artery blood flow increased by 89% (p< 0.005) whilst the HBF at the same time decreased by 32% (p< 0.005) in comparison to preocclusive HABF and HBF values. In sensory-denervated rats the resting HBF and PBF were increased by 23% (p< 0.05) and 34% (p< 0.05), respectively in comparison to the control HBF and PBF values. In this group the induction of the HABR increased the hepatic artery blood flow by only 55% (p< 0.05), whilst the HBF was reduced by 45% (p< 0.05). Pretreatment with CGRP 8-37 (CGRP receptor antagonist) and NK-1 but not NK-2 nor NK-3 receptor antagonists significantly reduced the HABF by 43% (p< 0.05) and 25% (p< 0.05) as compared to the HABF value in the control HABR group. These findings support the hypothesis that the hepatic artery buffer response induced by reduction of the portal inflow to the liver by 90% is partially mediated by activation of capsaicin-sensitive sensory fibers in the liver, probably due to local tissue ischemia and hypoxia. The observed vasodilation in the vascular bed of the hepatic artery is due to stimulation of CGRP and NK-1 receptors. PMID- 15795482 TI - Estimating parameters in the Rasch model in the presence of null categories. AB - A category with a frequency of zero is called a null category. When null categories are present in polytomous responses, then in the Rasch model for such responses, the thresholds that define the categories are inestimable with the commonly used joint maximum likelihood, marginal maximum likelihood, or standard conditional maximum likelihood estimation algorithms. The reason for this situation is that in principle, these estimation algorithms involve frequencies of each category. Andrich and Luo (2003) describe an algorithm in which the thresholds are reparameterized into their principal components and in which the estimate of any threshold is based on a function of the frequencies of all categories of the item rather than the frequency of a particular category. This algorithm works in the presence of null categories. However, in situations where the null categories are at the extremes of a set of categories, the estimates themselves can become too extreme. This paper describes a procedure in which the solution algorithm described by Andrich and Luo is further adapted in the presence of null categories by using their expected frequencies. The procedure is demonstrated with simulated and real data. PMID- 15795483 TI - Effect of item redundancy on Rasch item and person estimates. AB - One of the assumptions of many latent trait models is local independence. This assumption specifies that, after controlling for the underlying trait, item responses are independent. Given the lack of studies of model robustness against such violations, it appears that this assumption is frequently taken for granted. Therefore, this study investigated the robustness of Rasch item and person estimates with simulated data under varying number of items, sample sizes, and levels of item redundancy. Item and person reliabilities, the standard deviations of the person and item estimates, the root mean squared differences and mean signed differences among person and item estimates, the correlations between person estimates, and the percentage of person estimates shifting by more than .50 logits were used to evaluate the impact of item redundancy. Both norm and criterion-reference interpretations may be influenced by the imputation of redundancy into the data. However, it appears that the amount of redundancy needs to be considerable before such interpretations would be adversely impacted. Suggestions for further simulation research are provided. PMID- 15795484 TI - Measuring progress toward smoking cessation. AB - Measuring the effect of behavioral interventions is often limited to a single outcome variable for ease of analysis. In the case of low probability outcomes, this narrow focus may often result in Type II errors, reducing the likelihood of detecting an effect of an intervention. The development and use of a scale to measure progress toward the ultimate desired change in behavior might result in greater sensitivity to subtle, but important, effects of interventions. That possibility is illustrated by the development and exploratory testing of a scale meant to measure penetration into the process of change with respect to smoking cessation. The scale consists of a set of outcome indicators that are intended to represent the sequential steps that smokers go through in moving toward and ultimately giving up smoking. Rasch analyses indicate that the scale is coherent and merits further development. It seems likely that similar scales might be developed to assess progress toward change for many other behaviors that seem to require a gradual process of change that can be indexed by items representing discrete steps along the way. PMID- 15795485 TI - Daredevil barnstorming to the tipping point: new aspirations for the human sciences. AB - Aviation history provides an apt metaphor for the state of Rasch measurement practice, and its potential future. Flying was initially widely believed to be nothing but a spectacular and dangerous fad. Few saw in it any potential for the huge industry that it is today. The current state of Rasch measurement practice is quite akin to daredevil barnstorming in that the field is focused on isolated demonstrations of disconnected technical effects. Only when the analogues of air traffic control, airports, support staff, training programs, textbooks, and partner industries (hotels, restaurants) are in place will Rasch measurement come into its own as the technical medium of a widespread industry. The point at which current practice tips into a new paradigm depends on the realization of operationally validated theory in a supportive social context. The paper closes with speculations on what crossing Rasch measurement's tipping point might entail. PMID- 15795487 TI - A rank-ordering method for equating tests by expert judgment. AB - This paper describes a new method of comparing the raw mark scales on two tests using expert judgment. The two tests do not need to have any common items, nor to be taken by common groups of candidates. This study used scripts (i.e., the complete work of a candidate on the test) from England's National Curriculum Test for Reading at Key Stage 3 (14-year olds) in 2003 and 2004. Each member of a panel of 12 experts was given four packs each containing ten scripts--five scripts from each year's test. Marks and annotations from these scripts had been removed. Their task was to put the ten scripts into a single rank order, based on a holistic judgment of the level of performance exhibited in each. Because the design of the study linked scripts across judges and packs it was possible to construct a single latent trait of judged quality of performance. This was done using two different analytical methods: the Rasch formulation of Thurstone paired comparisons, and the Rasch Partial Credit model. Relating the two raw mark scales to the single latent scale allowed the two years' tests to be equated. The merits of using this standard-maintaining method as opposed to a standard-setting method in this particular context are discussed. PMID- 15795486 TI - Comparing Rasch analyses probability estimates to sensitivity, specificity and likelihood ratios when examining the utility of medical diagnostic tests. AB - INTRODUCTION: Medical diagnostic tests are evaluated based on measures of sensitivity (Sn), specificity (Sp), and likelihood ratios (LR). These procedures are limited in the event of a biased gold standard or missing data. Interpretations of these measures are frequently inappropriate. PURPOSE: The Rasch measurement model (RMM) was examined as a method to provide evidence of diagnostic test utility in order to overcome the limitations of Sn, Sp, and LR. METHODS: Patients suspected of a knee ligament tear (n = 825) were studied, by evaluating four diagnostic tests. The RMM probability estimates for each test were compared to estimates of Sn, Sp, and LR. RESULTS: The RMM provided probability estimates for the diagnosis that were comparable to likelihood ratios. These probability estimates correlated with the estimates of Sn, Sp, and LR. The RMM estimates were not affected by missing data. DISCUSSION: The RMM may provide an alternative means to study the utility of medical diagnostic tests to estimate the probability of disease presence/absence. PMID- 15795488 TI - Using the Rasch model to validate stages of understanding the energy concept. AB - In recent years, there have been efforts to bridge science education with developmental psychology to develop theories on students developing understanding of science concepts from elementary to high school and beyond. The present study intends to test one such theory on students developing understanding of the energy concept. The theory states that students develop understanding of the energy concept by going through the following qualitatively distinct stages: (a) energy as activity/work; (b) energy as sources/forms, (c) energy transfer, (d) energy degradation, and (e) energy conservation. Three classes, one each from 4th grade, 8th grade, and high school physics class (grades 10, 11, and 12), completed a performance assessment. Students' performances were scored based on three traits of energy understanding: attention capacity, qualitative relations, and quantitative relations; each of the traits was defined into five hierarchical levels consistent with the five stages of understanding the energy concept. The Many-Facet Rasch Measurement (MFRM) model was used to analyze the effects of rater scoring severity, students' stages of energy understanding (theta), and difficulties of energy understanding traits. Results show that there was a discontinuity among the stages of understanding the energy concept, supporting the theory on students developing the understanding of the energy concept. PMID- 15795489 TI - Spider silk proteins--mechanical property and gene sequence. AB - Spiders spin up to seven different types of silk and each type possesses different mechanical properties. The reports on base sequences of spider silk protein genes have gained importance as the mechanical properties of silk fibers have been revealed. This review aims to link recent molecular data, often translated into amino acid sequences and predicted three dimensional structural motifs, to known mechanical properties. PMID- 15795490 TI - Purification, characterization and sequence analyses of the extracellular giant hemoglobin from Oligobrachia mashikoi. AB - We purified an extracellular hemoglobin with the molecular mass of ca. 440 kDa from the whole homogenates of Oligobrachia mashikoi (phylum Pogonophora) by a one step gel-filtration. The preparation was pure to be crystallized. The P50 values of the hemoglobin and the fresh blood prepared from O. mashikoi were about 0.82 Torr and 0.9 Torr, respectively, which were much lower than the P50 value of human hemoglobin. However, the n values of the hemoglobin and the blood were about 1.2 and 1.1, respectively. Using the improved tricine SDS-PAGE, we could separate O. mashikoi hemoglobin into four kinds of the globin chains, A1, A2, B1 and B2, and succeeded for the first time in cloning and sequencing of the complete cDNA encoding B1 globin gene, in addition to A1, A2 and B2 globin genes in full length. We found that all globin genes have the extracellular signal sequences in each molecule and the distal His of the B1 globin chain is replaced to Gln. Finally, we constructed phylogenetic trees of the hemoglobins from Pogonophora, Vestimentifera and Annelida. PMID- 15795491 TI - Sperm-activating peptide induces asymmetric flagellar bending in sea urchin sperm. AB - Speract, a sperm-activating peptide (SAP) from sea urchin eggs, induces various sperm responses including a transient increase in the intracellular Ca2+ concentration. However, it has not been clarified how speract modulates sperm motility and whether it functions as a chemoattractant. To confirm the effect of speract on sperm motility, we observed the flagellar bending response to speract in sperm of Hemicentrotus pulcherrimus, in experiments using caged speract and a lighting system for a microscope newly developed with a power LED. We found that speract induces increases in curvature of swimming paths and changes flagellar bending shape to asymmetric. These facts show that speract directly regulates flagellar motility, and suggest that speract-induced increases in intracellular Ca2+ concentration play an actual role in regulation of the flagellar movement. PMID- 15795492 TI - Tissue-specific profile of DNA replication in the swimming larvae of Ciona intestinalis. AB - The cell cycle is strictly regulated during development and its regulation is essential for organ formation and developmental timing. Here we observed the pattern of DNA replication in swimming larvae of an ascidian, Ciona intestinalis. Usually, Ciona swimming larvae obtain competence for metamorphosis at about 4-5 h after hatching, and these competent larvae initiate metamorphosis soon after they adhere to substrate with their papillae. In these larvae, three major tissues (epidermis, endoderm and mesenchyme) showed extensive DNA replication with distinct pattern and timing, suggesting tissue-specific cell cycle regulation. However, DNA replication did not continue in aged larvae which kept swimming for several days, suggesting that the cell cycle is arrested in these larvae at a certain time to prevent further growth of adult organ rudiments until the initiation of metamorphosis. Inhibition of the cell cycle by aphidicolin during the larval stage affects only the speed of metamorphosis, and not the formation of adult organ rudiments or the timing of the initiation of metamorphosis. However, after the completion of tail resorption, DNA replication is necessary for further metamorphic events. Our data showed that DNA synthesis in the larval trunk is not directly associated with the organization of adult organs, but it contributes to the speed of metamorphosis after settlement. PMID- 15795493 TI - The carbonic anhydrase domain protein nacrein is expressed in the epithelial cells of the mantle and acts as a negative regulator in calcification in the mollusc Pinctada fucata. AB - Signals and organic matrix proteins secreted from the mantle are critical for the development of shells in molluscs. Nacrein, which is composed of a carbonic anhydrase domain and a Gly-X-Asn repeat domain, is one of the organic matrix proteins that accumulates in shells. In situ hybridization revealed that nacrein was expressed in the outer epithelial cells of the mantle of the pearl oyster Pinctada fucata. The recombinant nacrein protein inhibited the precipitation of calcium carbonate from a saturated solution containing CaCl2 and NaHCO3, indicating that it can act as a negative regulator for calcification in the shells of molluscs. Because deletion of the Gly-X-Asn repeat domain of nacrein had a significant effect on the ability of nacrein to inhibit the precipitation of calcium carbonate, it is conceivable that the repeat domain has a primary role in the inhibitory function of nacrein in shell formation. Together these studies suggest that nacrein functions as a negative regulator in calcification in the extrapallial space between the shell and the mantle by inhibiting the precipitation of CaCO3. PMID- 15795495 TI - Central connection of the optic, oculomotor, trochlear and abducens nerves in medaka, Oryzias latipes. AB - Medaka (Oryzias latipes) is one of the few vertebrate experimental animals in which inbred lines have been established. It is also a species that has advanced in genetic studies in a manner comparable to zebrafish. This fish is therefore a good model for studying functional organization of the nervous system, but anatomical analysis of its nervous system has been limited to embryonic stages. In the present study, we investigated anatomy of cranial nerves in adult fish focusing on the visual function, using an inbred strain of medaka. Cranial nerves of medaka were labeled using biocytin, revealing a central distribution of retinofugal terminals, retinopetal neurons, and oculomotor, trochlear and abducens motor neurons. The optic nerve of the adult medaka was of a complete decussation type. Retinofugal terminals were located in 8 brain nuclei, the suprachiasmatic nucleus, nucleus pretectalis superficialis, nucleus dorsolateralis thalami, area pretectalis pars dorsalis (APd), area pretectalis pars ventralis (APv), nucleus of the posterior commissure (NPC), accessory optic nucleus, and the tectum opticum. Retinopetal neurons were identified in 6 brain nuclei, the ganglion of the terminal nerve, preoptic retinopetal nucleus, nucleus dorsolateralis thalami, APd, APv, and NPC. The oculomotor neurons were mostly labeled ipsilaterally and were located dorsomedially, abutting the fasciculus longitudinalis medialis in the mesencephalon. The trochlear nucleus was located contralaterally and dorsolaterally adjacent to the fasciculus longitudinalis medialis in the mesencephalon. The abducens nucleus was located ipsilaterally in a ventrolateral part of the rhombencephalic reticular formation. These results, generally similar to those in other teleosts, provide the basis for future behavioral and genetic studies in medaka. PMID- 15795494 TI - In vivo effects of a recombinant molt-inhibiting hormone on molt interval and hemolymph ecdysteroid level in the kuruma prawn, Marsupenaeus japonicus. AB - In order to determine the function of molt-inhibiting hormone (MIH) in vivo, we examined the effects of injecting of a recombinant MIH on the molt interval and hemolymph ecdysteroid level in the kuruma prawn, Marsupenaeus japonicus. The injection of recombinant MIH significantly prolonged the molt interval (9.0 +/ 0.4 days in the control group, 9.5+/-0.5 days in the 2500 ng/g-body weight/injection-group, mean+/-SD), and significantly decreased the hemolymph ecdysteroid level (ratio of levels between after and before injection: 1.94+/ 1.09 in the control and 1.28+/-0.39 in the 3000 ng/g-body weight/injection-group, mean+/-SD). These results conclusively show the inhibitory effects of MIH on molting in vivo. PMID- 15795496 TI - Brain-derived neurotrophic factor promotes neurite growth and survival of antennal lobe neurons in brain from the silk moth, Bombyx mori in vitro. AB - This study was conducted to investigate effects of brain-derived neurotrophic factor on the neurite growth and the survival rate of antennal lobe neurons in vitro, and secretion of brain-derived neurotrophic factor-like neuropeptide from brain into hemolymph in the silk moth, Bombyx mori. In primary culture of antennal lobe neurons with brain-derived neurotrophic factor, it promoted both a neurite extension of putative antennal lobe projection neurons and an outgrowth of branches from principal neurites of putative antennal interneurons with significance (p<0.05). Brain-derived neurotrophic factor also increased significantly a survival rate of antennal lobe neurons (p<0.05). Results from immunolabeling of brain and retrocerebral complex, and ELISA assay of hemolymph showed that brain-derived neurotrophic factor-like neuropeptide was synthesized by both median and lateral neurosecretory cells of brain, then transported to corpora allata for storage, and finally secreted into hemolymph for action. These results will provide valuable information for differentiation of invertebrate brain neurons with brain-derived neurotrophic factor. PMID- 15795497 TI - Phylogeny and evolution of butterflies of the genus Parnassius: inferences from mitochondrial 16S and ND1 sequences. AB - Phylogenetic relationships among species of the genus Parnassius and its related taxa were analyzed by comparing nucleotide sequences of mitochondrial 16S ribosomal RNA (504 sites) and NADH-dehydrogenase subunit 1 (469 sites). In the phylogenetic trees, Parnassius was found to be most closely related to Hypermnestra helios, whereas Archon apollinus, which has been classified in the tribe Parnassiini together with Parnassius and Hypermnestra, was more closely related to members of the tribe Zerynthiini. Within the Parnassius clade, six major clades corresponding to species groups were well supported, although the phylogenetic relationships among them were not clear. Although the results of the present study were in agreement with those of a previous phylogenetic study based on mitochondrial NADH-dehydrogenase subunit 5 sequences, our study strongly supported a close relationship between Parnassius and Hypermnestra, which was not well supported in the previous study. PMID- 15795498 TI - Phylogenetic relationships of the tribe Paini (Amphibia, Anura, Ranidae) based on partial sequences of mitochondrial 12s and 16s rRNA genes. AB - Partial sequences of mitochondrial 12S and 16S rRNA genes from 19 Asian frog species of the tribe Paini (Ranidae, Dicroglossinae) allowed a first molecular study of the phylogenetic relationships of this tribe. This analysis confirmed that this tribe is a monophyletic group, but suggested relationships did not agree with previous generic classification of this clade based on morphology. Two major clades were recognized within the Paini. For one of them, the generic name Quasipaa is available. Phylogenetic relationships within the other group are not yet fully clarified and need further study. PMID- 15795499 TI - An atrial membrane in the colonial ascidian, Ritterella tokioka Kott, 1992 (Urochordata: Ascidiacea) from Sagami Bay. AB - An atrial membrane, similar to that previously reported (as a placental membrane) in Placentela crystallina Redikorzev, 1913, has been found in the holotype and other specimens of another aplousobranch ascidian, Ritterella tokioka Kott, 1992 (=R. pedunculata Tokioka, 1953). In serial sections, the membrane is seen to be an extensive fold of the atrial epithelium over the rectum that projects into the atrial cavity and supports developing embryos over its outer surface. The similar states in the atrial membrane seen in the two species are attributable to homoplasy, rather than homology. PMID- 15795500 TI - Two new species of diplosoma (Ascidiacea: Didemnidae) bearing prokaryotic algae prochloron from Okinawajima (Ryukyu Archipelago, Japan). AB - Two new species of didemnid ascidians, Diplosoma ooru sp. nov. and Diplosoma simileguwa sp. nov., are described from coral reefs on Okinawajima (Ryukyu Archipelago, Japan). These two species form green colonies, having a symbiotic association with a prokaryotic alga Prochloron sp. The former species was found at the reef edges in the subtidal zone and the latter was found in a shallow reef lagoon. In these species, the colonies are thinner and the zooids are smaller than those of any other Prochloron-bearing Diplosoma species so far described. Moreover, each of the present new species has a unique combination of stigmatic numbers: 5 stigmata in the first and third rows, 6 in the second row, and 4 in the fourth in D. ooru; 4 stigmata in the first and third rows, 5 in the second row, and 3 in the fourth in D. simileguwa. In both of the new species, the retractor muscle emerges from the underside of the thorax. Larval morphology of D. ooru is also described. PMID- 15795502 TI - [Experimental study on treatment planning verification using a positron emitting (10)C beam for heavy-ion radiotherapy.]. AB - An advantage of heavy-ion therapy is its good dose concentration. A limit for full use of this desirable feature comes from range ambiguities in treatment planning. The treatment planning is based on X-ray CT measurements, and the range ambiguities are mainly due to an error in calibration of the CT number. The heavy ion ranges are related to electron density of the medium while the CT numbers are defined using the X-ray attenuation coefficient. The range verification method using positron emitter beams has been developed to reduce the range ambiguities. In this verification, probing beams of positron emitters are implanted into the tumor, and pairs of annihilation gamma rays are detected with a positron camera. This paper demonstrates an application to verify treatment planning. Here the treatment planning was made on a head phantom and the ranges estimated from the CT-number were compared with the ranges measured with the positron camera. As a result, disagreements were detected between the planned ranges and the measured ones; there were 1.6 mm at maximum. The disagreements were due to an error of transformation of CT-number to range for the phantom material in the water column depth-dose measurement. The disagreements could be lowered to 0.4 mm by using the calibrated water-equivalent lengths. It was confirmed that the range verification system has a designed measurement accuracy of 1 mm and is useful for verifying irradiation fields on heavy-ion radiotherapy. PMID- 15795501 TI - Mammographic x-ray spectra measured by Compton spectroscopy using a high resolution Schottky CdTe detector. AB - Analysis of x-ray spectra is important for quality assurance (QA) and quality control (QC) of radiographic systems. This is especially true for mammographic x ray imaging systems which require low-contrast detectability. Under clinical conditions, the measurement of diagnostic x-ray spectra is difficult because of pulse pile-up due to the high fluence rate of incident x-ray photons. However, it is difficult to set a long source-to-detector distance to reduce the number of photons detected per unit time for mammographic x-ray units. Compton spectroscopy is a suitable tool to deal with this problem. Detection of 90-degrees scattered photons only, energy correction and reconstruction are easy using the Klein Nishina formula. However, x-ray spectra measured by this method have lower energy resolution, because of the geometrical irradiation angle or electron movement in the scatterer. Moreover, spectra measured with a compound semiconductor detector, such as a high resolution Schottky CdTe detector like we used, are distorted by the detector response, which is based on detecting x-ray photon interactions and charge carrier trapping in the semiconductor crystal. While the distortion of spectra caused by the response can be easily corrected by applying a stripping procedure, it is very difficult to reconstruct the broad spectra measured by Compton spectroscopy as sharp spectra such as obtained when directly measured. Some complicated reconstruction algorithms have been reported to fit the shape of spectra obtained by the Compton spectroscopy to sharp standard spectra. However, for QA / QC of the radiographic system, it is not necessary to correct the spectra sharply if the spectral broadening is at a tolerable level and the properties of the broad spectra acquired by the Compton spectroscopy agree with those of the sharp spectra measured directly; i.e. evaluations are necessary only for estimation of spectral shape. In this paper, we compared attenuation curves calculated using Hubbell's attenuation coefficients to estimate the coincidence or difference of spectra measured by the Compton spectroscopy and directly measured in the primary beam. Our results showed that the attenuation curves acquired from the reconstructed spectra measured by the Compton spectroscopy agreed with that acquired from corrected spectra that were directly measured. Moreover, the attenuation curves acquired from the spectra actually measured by adding aluminum attenuators agreed with theoretically calculated curves. PMID- 15795503 TI - [Comparative evaluation of absorbed dose and image quality in X-ray head CT scanning for hospital facilities in Gunma.]. AB - Recently, the number of scans for X-ray computed tomography (CT) examinations has been rising due to the wide-spread use of multi-slice CT (MSCT) scanners. There is a concern that the total medical exposures will be increased by these examinations. In order to lower exposures, routine parameters for head CT examinations done at several hospitals in Gunma were investigated. In this study, the computed tomography dose index (CTDI(100, C)), noise, and low contrast resolution were measured. The CTDI(100, C) for all the hospitals exceeded the guideline (40mGy) suggested by the Japan Association of Radiological Technologists (JART). Low contrast resolution showed the coefficient of variation of +/-5% between hospitals. In conclusion, it was proposed that the technologists should reconsider the parameters of the head X-ray CTs, in cases where their output dose far exceeds the standard. PMID- 15795504 TI - [An analysis of a new method for estimating the range of hand motion using tele rehabilitation.]. AB - We compared estimated axes obtained from the upper arm outline, which changes during movement, with the skeleton axis obtained from an x-ray image in terms of range of motion. Statistical analysis showed that the estimated axes corresponded to the skeleton axis in each range of motion of the hand. The analysis was superior to the conventional approach. PMID- 15795505 TI - Growth in extremely low birth weight infants up to three years. AB - OBJECTIVE: To evaluate postnatal growth of extremely low birth weight infants (ELBW, <1,000 g) until 36 months of corrected age (CA), and to relate growth outcome to anthropometric parameters at birth, sex, fetal growth status (small or appropriate for gestational age--SGA/AGA), period of admission and major perinatal events. STUDY DESIGN/METHOD: Weight (Wt), height (Ht) and head circumference (HC) were assessed in 159 ELBW infants. Data were standardized with Z-scores following Usher and McLean and Sempe growth curves. Uni- and multivariate statistical analysis were performed. RESULTS: The mean birth weight was 851.2+/-116.5 g. Z-scores decreased from birth to term, at a deeper rate for AGA than for SGA infants (p<0.005 for Ht, Wt, and HC). Between term and 36 months, growth was better in SGA compared with AGA infants (p=0.003 for Ht). Multivariate analysis showed that anthropometric parameters at birth were positive determinants for Wt, Ht and HC at term, and also for Wt and Ht at 36 months CA (Z-scores). Oxygen therapy after 36 weeks of post-conceptional age was a negative determinant influencing growth at 36 months CA (Z-scores). CONCLUSIONS: Significant catch-up growth took place between birth and 36 months, which was greater for SGA than for AGA infants. Anthropometric parameters at birth and oxygen therapy at 36 weeks post-conceptional age are the main predictive factors for growth at 36 months CA. PMID- 15795506 TI - Brain cell function during hypoxemia in near-term lambs: a near-infrared spectroscopy study. AB - BACKGROUND: Sufficient O2 supply to the brain is necessary for adequate cerebral energy metabolism, function and growth. OBJECTIVES: To elucidate the relation between changes in cerebral arterial O2 content and cerebral O2 supply and changes in the oxygenation state of cerebral hemoglobin, and to determine whether concentration changes in oxyhemoglobin (DeltacO2Hb), deoxyhemoglobin (DeltacHHb), and cerebral arterial oxygenation (DeltacHbD; the difference between DeltacO2Hb and DeltacHHb), and cerebral blood volume (DeltaCBV) can be used to assess the decline in brain cell function during hypoxemia in lambs born near term. METHODS: 17 preterm lambs were delivered at a mean gestational age of 133 days. Decreases in cerebral arterial oxygen content were induced by a stepwise reduction in inspired oxygen concentration. Mean values of all continuous variables were calculated over the last 180 s of each hypoxemic level. Cerebral arterial blood gases were analyzed at the end of each level to calculate cerebral arterial O2 content and cerebral O2 supply. RESULTS: Changes in cerebral arterial O2 content and cerebral O2 supply were positively linearly related with DeltacO2Hb and DeltacHbD, and negatively with DeltacHHb and the concentration changes in total hemoglobin. Electrocortical brain activity remained stable until the cO2Hb and cHbD decreased to >3.0 +/- 0.9 and >8.1 +/- 1.9 (mean +/- SD) micromol/100 g, respectively, and cHHb and CBV increased to >4.3 +/- 1.7 and 1.37 +/- 0.48 ml/100 g, respectively, as compared to baseline. CONCLUSIONS: Changes in cerebral arterial O2 content and cerebral O2 supply are adequately reflected by changes in the oxygenation state of cerebral hemoglobin. Concentration changes in DeltacO2Hb, DeltacHHb, DeltacHbD and DeltaCBV can be used to assess the decline in brain cell function during hypoxemia in lambs born near term. PMID- 15795507 TI - What causes increased stroke mortality in patients with prestroke dementia? AB - BACKGROUND AND PURPOSE: In patients with dementia, the incidence of stroke is higher and strokes are more severe and lethal. The purpose of this population based study was to describe in what way previous dementia affects mortality in stroke patients. METHODS: Subjects were all persons > or =65 years old who had a first-ever stroke during 1 year (n = 327). The prestroke dementia (PSD) diagnosis was made at the time of the stroke diagnosis using data from next of kin and from patient records. Patients were followed prospectively and causes of death were evaluated. RESULTS: The 28-day case fatality was 44% for PSD patients and 15% for non-PSD patients. Corresponding ratios at 1 year were 71 and 36%, respectively. Twenty-eight percent of the PSD patients had a new stroke during the first year, compared to 8% of the non-PSD patients. More patients in the PSD group died as a direct or indirect consequence of their stroke. Multivariate analysis showed that PSD, in addition to age, atrial fibrillation and stroke severity, was an independent predictor of mortality. CONCLUSIONS: The PSD patients more often had a stroke-related death, even when we adjusted for a number of other factors. The cause for this is most likely multifactorial, including an increased tendency to contract complications in the acute phase, and iatrogenic causes. The brain of the PSD patients may also be frailer and more susceptible to ischemic or hemorrhagic damage than the nondemented brain. PMID- 15795508 TI - What has diffusion imaging in animals told us about diffusion imaging in patients with ischaemic stroke? AB - BACKGROUND AND PURPOSE: In acute ischaemic stroke, the amount (and type) of cellular damage underlying diffusion-weighted imaging (DWI) lesion appearances is unclear. We summarized all information from experimental studies of DWI in focal ischaemia models. METHODS: We systematically reviewed all published studies of DWI in focal ischaemic stroke models. We extracted key experimental details to determine CORRELATIONS between histological features and DWI lesion characteristics. RESULTS: Of 141 potentially eligible papers (including more than 2,817 animals, mostly rats), details of key experimental methods were unfortunately often omitted. Consistent findings amongst high-quality studies with blinded analysis included: neuronal damage persists or progresses despite early DWI lesion 'normalisation'; the apparent diffusion coefficient is not very sensitive to the amount of neuronal damage; the 'brighter' the DWI lesion, the greater the neuronal damage; and the DWI lesion may reflect glial more than neuronal changes. Anaesthesia and fixation techniques may inadvertently affect these findings. CONCLUSIONS: The relationship between cellular damage and DWI lesion appearance, particularly recovery patterns in reperfusion experiments, remains imprecise. Key experimental details could be reported more completely and consistently. Potential problems from repeated anaesthetics need to be addressed. Early DWI lesion 'recovery' in acute stroke patients may largely reflect glial rather than neuronal 'recovery'. PMID- 15795509 TI - Lack of telomere shortening with age in mouse resting zone chondrocytes. AB - BACKGROUND AND AIM: Telomeres are hexameric repeat sequences that flank eukaryotic chromosomes. The telomere hypothesis of cellular aging proposes that replication of normal somatic cells leads to progressive telomere shortening which induces replicative senescence. Previous studies suggest that growth plate chondrocytes have a finite proliferative capacity in vivo. We therefore hypothesized that telomere shortening in resting zone chondrocytes leads to replicative senescence. METHOD: To test this hypothesis we compared the telomere restriction fragment (TRF) length of Mus casteneus at 1, 4, 8, and 56 weeks of age. RESULTS AND CONCLUSIONS: We found that TRF length did not diminish measurably with age, suggesting that telomere shortening in resting zone chondrocytes is not the mechanism that limits proliferation of growth plate chondrocytes in vivo. PMID- 15795510 TI - Ghrelin in growth and development. AB - Exogenous administration of ghrelin increases caloric intake and stimulates growth hormone (GH) secretion, two effects that are mediated through binding of ghrelin to the GH secretagogue receptor (GHS-R). In addition, ghrelin is thought to inhibit adipogenesis by GHS-R-independent mechanisms. In adults, ghrelin is mainly produced by the stomach. In contrast, in the fetal and early postnatal period, ghrelin gene expression is abundant in the pancreas but not in the stomach. While knockout animal studies demonstrate that ghrelin is not required for perinatal development under normal nutritional conditions, the characteristics of ghrelin metabolism during fetal development suggest that ghrelin could contribute to the programming of mechanisms involved in energy balance, such as beta-cell maturation, orexigenic pathways and adipogenesis. In humans, ghrelin concentrations progressively decrease during childhood and adolescence, as well as with advancing puberty. In adolescents, similar to adults, ghrelin concentrations are inversely related to body mass index and to circulating insulin. One notable exception is the presence of elevated ghrelin concentrations in subjects with Prader-Willi syndrome, raising the possibility that ghrelin could be part of the etiology of excess food intake in this condition. These data raise a number of fascinating questions on the potential physiologic role of this hormone during growth and development. PMID- 15795511 TI - Manifestation, management and molecular analysis of candidate genes in two rare cases of thyrotoxic hypokalemic periodic paralysis. AB - BACKGROUND: Hypokalemic periodic paralysis as a complication of thyrotoxicosis (THypoKPP) is common in Asians but not well recognized in Western countries or pediatric patients, where most cases are due to the familial variant (FHypoKPP). Ion channel gene mutations may underlie these diseases. We describe the first pediatric and a rare adult Caucasian case of THypoKPP in Finland. METHODS: Manifestation and management of two THypoKPP cases. We studied for possible mutations in KCNE3, KCNJ2, SCN4A and CACNA1S genes. RESULTS: A 15-year-old Vietnamese boy presented with sudden-onset paralysis and severe hypokalemia, 1.8 mmol/l. The case was first regarded as FHypoKPP, but thyroid function testing revealed a suppressed TSH and highly elevated FT4. A 37-year-old Caucasian male presented with acute tetraparesis. His plasma potassium was only 1.4 mmol/l. Treatment with carbimazole had been initiated two weeks earlier, but FT4 was still elevated. No mutations in KCNE3, KCNJ2, SCN4A or CACNA1S genes were detected. CONCLUSIONS: THypoKPP is a potentially life-threatening condition which bares many similarities with FHypoKPP. THypoKPP is rare in Western countries but should be considered in sudden-onset paralysis, independently of age and especially in males. Mutations in ion channel candidate genes did not underlie the disease in the present cases. PMID- 15795512 TI - Height, weight, IGF-I, IGFBP-3 and thyroid functions in prepubertal children with attention deficit hyperactivity disorder: effect of methylphenidate treatment. AB - OBJECTIVE: To investigate if there are any disease-related or methylphenidate induced aberrations in growth parameters, growth hormone insulin-like growth factor (IGF)-I, IGFBP-3 axis and the thyroid function tests in children with attention deficit hyperactivity disorder (ADHD). METHODS: Newly diagnosed and untreated prepubertal children with ADHD were longitudinally followed before and approximately every 4 months after methylphenidate treatment for up to 16 months. Height SDS, weight SDS, BMI SDS, serum GH, IGF-I, IGFBP-3, T4, free T4, T3, and TSH were measured at each visit. RESULTS: All of the examined parameters were within normal limits for age before treatment. Methylphenidate treatment did not significantly affect SDS of height, weight, BMI, IGF-I and IGFBP-3 in the long run. Serum T4 and free T4 levels showed modest reductions within normal limits in a time-dependent manner. CONCLUSIONS: Prepubertal children with ADHD had normal height, weight, BMI, serum IGF-I and IGFBP-3 and thyroid functions. Methylphenidate treatment had no sustained effects on growth parameters, IGF-I and IGFBP-3 during the follow-up period of this study. However, it caused a mild decrease in total and free T4 which may warrant further monitoring. PMID- 15795513 TI - Reduced bone mineral density and radial bone growth in young rabbits treated with dexamethasone eye drops. AB - OBJECTIVE: To investigate the effect of dexamethasone eye drops on bone metabolism in newborn rabbits. METHODS: Thirty-four 3-week-old rabbits had unilateral clear lens extraction and were randomized into three groups. Postoperatively, group 1 received high-dose and group 2 low-dose dexamethasone eye drops (average doses 0.27 and 0.10 mg/kg body weight/day, respectively). These rabbits also received a postoperative subconjunctival injection of betamethasone. Group 3 (control) received vehicle eye drops only. After 8 weeks of treatment, all animals were killed and the left femurs were isolated and subjected to peripheral quantitative computerized tomography (pQCT) and dual X ray absorptiometry (DXA) analyses. RESULTS: DXA showed that rabbits treated with either a high or low dose of dexamethasone eye drops had significantly reduced areal bone mineral density (BMD), area and total bone mineral content (BMC) of the femur. Measurements with pQCT demonstrated a dose-dependent reduction in cortical BMC, cortical volumetric BMD and cortical area. These effects were associated with an inhibition of radial femur growth, cortical thickness and periosteal and endosteal circumferences. CONCLUSION: Dexamethasone eye drops have systemic effects affecting several bone parameters in young rabbits. Any long term systemic effects of ocular glucocorticoids need to be further studied. PMID- 15795514 TI - Hereditary paraganglioma/pheochromocytoma and inherited succinate dehydrogenase deficiency. AB - Mitochondrial complex II, or succinate dehydrogenase, is a key enzymatic complex involved in both the tricarboxylic acid (TCA) cycle and oxidative phosphorylation as part of the mitochondrial respiratory chain. Germline succinate dehydrogenase subunit A (SDHA) mutations have been reported in a few patients with a classical mitochondrial neurodegenerative disease. Mutations in the genes encoding the three other succinate dehydrogenase subunits (SDHB, SDHC and SDHD) have been identified in patients affected by familial or 'apparently sporadic' paraganglioma and/or pheochromocytoma, an autosomal inherited cancer susceptibility syndrome. These discoveries have dramatically changed the work-up and genetic counseling of patients and families with paragangliomas and/or pheochromocytomas. The subsequent identification of germline mutations in the gene encoding fumarase--another TCA cycle enzyme--in a new hereditary form of susceptibility to renal, uterine and cutaneous tumors has highlighted the potential role of the TCA cycle and, more generally, of the mitochondria in cancer. PMID- 15795515 TI - Roles of nitric oxide and oxidative stress in the regulation of blood pressure and renal function in prehypertensive Ren-2 transgenic rats. AB - AIMS: The present study was performed to evaluate the role of nitric oxide (NO) and its interaction with superoxide anion (O2-) in the regulation of blood pressure (BP) and renal function during the developmental phase of hypertension in Ren-2 transgenic rats (TGR). The first aim was to compare BP and renal functional responses to acute NO synthase (NOS) inhibition achieved by intravenous (i.v.) infusion of Nomega-nitro-L-arginine-methyl ester (L-NAME) in prehypertensive heterozygous TGR and in transgene-negative Hannover Sprague Dawley (HanSD) rats. The second aim was to evaluate whether scavenging of O2- by infusion of the superoxide dismutase mimetic tempol increases NO bioavailability which therefore should augment BP and renal functional responses to L-NAME. METHODS: Rats were anesthetized, prepared for clearance experiments and BP and renal functional responses were evaluated in response to i.v. L-NAME administration (20 microg.100 g(-1).min(-1)) without or with tempol pretreatment (i.v., 300 microg.100 g(-1).min(-1)). In renal cortical tissue, nitrotyrosine protein expression was assessed by immunoblotting as marker of O2- production and urinary 8-epi-PGF(2alpha) excretion as marker of intrarenal oxidative stress was assessed by enzyme immunoassay. RESULTS: BP, glomerular filtration rate (GFR), renal plasma flow (RPF) and sodium excretion were similar in TGR and HanSD. L NAME infusion induced greater increases in BP in TGR than in HanSD (+42 +/- 4 vs. +25 +/- 3 mmHg, p < 0.05). In the absence of a significant change in GFR, L-NAME caused similar decreases in RPF (-32 +/- 6 and -25 +/- 4%, p < 0.05) in TGR and HanSD. Despite significantly higher renocortical expression of nitrotyrosine and urinary 8-epi-PGF2alpha excretion in TGR than in HanSD, pretreatment with tempol did not augment the rise in BP and the decrease in RPF induced by L-NAME. CONCLUSIONS: The greater BP response to L-NAME in TGR suggests that prehypertensive TGR exhibit an enhanced NO activity in the systemic vasculature as compared with HanSD. Despite increased intrarenal oxidative stress in TGR, the dependency of the intrarenal vascular tone on NO appears to be similar in TGR and HanSD. The lack of a compensatory increase in renal NO activity may partially account for the enhanced renal vascular response to ANG II present in TGR. PMID- 15795516 TI - Nutritional and inflammatory markers in the prediction of mortality in Chinese hemodialysis patients. AB - BACKGROUND/AIMS: The prevalence of cardiovascular disease and mortality rate is relatively low in Chinese dialysis patients. This study aimed to evaluate the predictive value of nutritional and inflammatory markers in Chinese hemodialysis patients. METHODS: A total of 158 patients (70 men and 88 women, age 59.9 +/- 13.2 years) were studied. Nutritional and inflammatory markers, including subjective global assessment (SGA), insulin-like growth factor-1, albumin, tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6, serum amyloid A (SAA), and C reactive protein (CRP), were measured. These patients were followed up until April 2004 (36 months) to determine the incidence and causes of death. RESULTS: SGA (p = 0.001), IL-1beta (p = 0.032), SAA (p = 0.031), IL-6 (p = 0.001) and CRP (p < 0.001) were found to be significant predictors of mortality. After adjusting with age, sex, diabetes, coronary artery disease, Kt/Vurea, and duration on dialysis, CRP (odds ratio = 4.58; p = 0.038) and SGA (odds ratio = 6.57; p = 0.004) remained the independent predictors of mortality. The adjusted mortality rate was highest for patients with a high CRP level and malnutrition (assessed by SGA). CONCLUSIONS: SGA and CRP levels are the most significant predictors of mortality in Chinese dialysis patients. Chinese dialysis patients with a high CRP level tend to be at higher risk of mortality only if they are malnourished. PMID- 15795517 TI - All-trans-retinoic acid inhibits the development of mesangial proliferative glomerulonephritis in interleukin-6 transgenic mice. AB - All-trans-retinoic acid (ATRA), a vitamin A derivative, was reported to suppress the interleukin-6 (IL-6) production and to downregulate the IL-6 receptor (IL-6R) and/or its signal transducer glycoprotein 130. We investigated the in vivo antinephritic effect of ATRA on IL-6 transgenic mice which had developed mesangial proliferative glomerulonephritis (PGN) as well as its in vitro inhibitory effect on the proliferation of rat mesangial cells. In vivo experiments on IL-6 transgenic mice showed that ATRA administration suppressed proteinuria and hematuria and reduced the IL-6 concentrations; furthermore, histological examination demonstrated that it improved PGN. In vitro experiments using rat mesangial cells demonstrated that ATRA inhibited cell growth in a dose dependent manner within a range from 10(-4) to 10(-6) M. This inhibition by ATRA was partially counteracted by the addition of IL-6. RT-PCR assay results showed that ATRA also reduced IL-6R, but not the glycoprotein 130 expression in mesangial cells. These findings indicate that, by blocking of the IL-6 function, ATRA may be therapeutically effective in PGN. PMID- 15795518 TI - Protective effect of L-arginine intake on the impaired renal vascular responses in the gentamicin-treated rats. AB - The purpose of this study was to investigate the effect of gentamicin (100 mg/kg/day, i.p.) treatment on endothelium-dependent and -independent vasodilation in isolated perfused rat kidney, and the effect of amino acid L-arginine (in the drinking water, 2.25 g/l) on renal dysfunction induced by gentamicin. When gentamicin-treated groups were compared with the control group, it was observed that BUN and creatinine levels increased significantly. Also, the relaxant responses induced by acetylcholine, sodium nitroprusside and pinacidil decreased. Histopathological examination indicated acute tubular necrosis in this group. In animals treated with gentamicin together with L-arginine, there was a significant amelioration in the BUN and creatinine levels. The vasodilator responses were similar to those of the control group. Histopathological examination indicated only hydropic degeneration in tubular epithelium of kidney. Co-administration of L-NG-nitroarginine methyl ester (L-NAME) (112.5 mg/l), an inhibitor of nitric oxide synthase, and L-arginine to rats treated with gentamicin did not change the protective effect of L-arginine. In rats receiving L-NAME alone, the level of BUN and creatinine and vasodilation to acetylcholine were not significantly different when compared to those of the control group, while relaxant responses to sodium nitroprusside and pinacidil were increased. These results suggest that gentamicin leads to an impairment in vascular smooth muscle relaxation in addition to acute tubular necrosis in the rat kidney. Supplementation of L-arginine has an important protective effect on gentamicin-induced nephropathy. PMID- 15795519 TI - Reduced cyclooxygenase involvement in vascular endothelial function in rat renal transplantation. AB - BACKGROUND: Cardiovascular disease is a major cause of death following renal transplantation. Mechanisms leading to vascular dysfunction outside the transplanted organ involve common risk factors such as hypertension, hypercholesterolemia, proteinuria, but immune-mediated factors may also be involved. We hypothesized that transplantation-associated risk factors are involved in the development of vascular dysfunction following renal transplantation. METHODS: Vascular function was studied in Fisher to Lewis allografts. Lewis to Lewis syngrafted rats served as controls. All rats received cyclosporin A for 10 days. Allografts were treated with ACE inhibition or AT1 receptor blockade or left untreated. After 34 weeks, aorta rings were studied for contractile and dilator responses in the presence or absence of L-NMMA and/or indomethacin. Tissue sections were immunostained for COX-1 and COX-2. RESULTS: In contrast to syngrafts and treated allografts, untreated allografts developed proteinuria and hypercholesterolemia. In aortic rings, NOS inhibition similarly increased contractile responses and decreased dilator responses in syngrafts and allografts, indicating comparable NO pathways. In contrast, indomethacin affected contractile and dilator responses in syngrafts, but not in treated and untreated allografts, indicating absence of COX-derived prostanoids in control over vascular tone in allografts. This was in line with immunohistologic analysis demonstrating reduced aortic COX-2 expression in allografts. COX-1 expression was unaltered. Interestingly, RAS blockade quantitatively increased endothelium dependent dilation without qualitatively altering COX function and expression. CONCLUSION: Involvement of COX-derived prostaglandins in vascular endothelial function outside the transplanted organ is strongly diminished after allogeneic renal transplantation. RAS blockade improves common cardiovascular risk factors and endothelium-dependent dilation, but fails to restore prostaglandin function. PMID- 15795520 TI - Evaluation of the results of endoscope-assisted acoustic neuroma surgery through posterior fossa approach. AB - The objective of this study was to evaluate the results of endoscope-assisted acoustic neuroma surgery in posterior fossa approach. Between 1996 and 2002, 60 consecutive patients with acoustic neuroma were operated via the retrosigmoid suboccipital approach. Standard 4-mm sinus endoscopes at different angles were used during the surgeries either for inspection or tumor endoscopic dissection. Clinical parameters and treatment outcome were evaluated retrospectively. Tumor sizes were small, medium and large in 46.6, 45 and 8.3% of the patients, respectively. The hearing preservation rate, which did not correlate with tumor size (p > 0.05), was 24.4%. The need for facial reanimation surgery, which was needed in 5% of patients, was significantly higher in the large tumors than in the small and medium tumors (p < 0.001). Cerebrospinal fluid fistula rate, which was not related to tumor size (p > 0.05), was 13.3%. Tumor recurrence or residual tumor was not encountered at all. In conclusion, endoscopes give accurate information about the relationship between the tumor and the adjacent structures and help control the fundus of the internal auditory canal to ensure complete tumor removal. It is also helpful in visually verifying the continuity of the facial and cochlear nerves. The use of endoscopes does not appear to increase the hearing preservation rate, but is very helpful in complete tumor removal in the posterior fossa approach. PMID- 15795521 TI - Identification of adenosine receptor subtypes expressed in the human endothelial like ECV304 cells. AB - Adenosine, acting through its receptors, is a potent endogenous regulator of endothelial cells. The cultured endothelial cells expressing adenosine receptors are thus important for elucidation of molecular mechanism of adenosine functions in these cell systems. Therefore, identification of adenosine receptors in the human ECV304 cell line derived from a human umbilical vein endothelial cell culture was performed. RT-PCR experiments revealed that ECV304 cells express mRNAs for A1 and A2B adenosine receptors. The expression of mRNA for A2A adenosine receptor was not in a significant level and that for A3 adenosine receptor was not detected. The binding study of ECV304 cell membrane fractions using various radiolabeled ligands for adenosine receptors indicated the presence of A1 adenosine receptors 245 fmol/mg of membrane proteins, but the specific binding for A2A and for A3 adenosine receptors were found to be negligible. The functional expression of A1 and A2B adenosine receptors in ECV304 cells was detected by assays for adenosine-3',5'-cyclic monophosphate and for extracellular signal-regulated kinase, but that of A2A adenosine receptors was not confirmed under the assay conditions employed. In conclusion, this study presented evidence for functional A1 and A2B adenosine receptors in human endothelial-like ECV304 cells, indicating that ECV304 cells can be a good model for the study of adenosine receptors, especially for A2B adenosine receptor, in endothelial cells. PMID- 15795524 TI - Gag-p6 Tsg101 binding site duplications in maternal-infant HIV infection. AB - Prevalence and patterns of HIV p6 duplications in HIV-1 mother-to-baby transmission are examined. Resistance genotyping was performed in a multisite U.S. study of antiretroviral resistance in vertical transmission. Sequence data were used in secondary analyses of HIV genetic variation. Two hundred sixty HIV viral RNA samples from HIV-infected pregnant women and their infants were analyzed with a commercial resistance genotyping kit. Chromatograms were examined for variability in the 3' region of gag. From 103 mother-baby sets, 190 samples gave readable p6 sequence. Of 103 mother-baby sets, 20 (19%) showed duplication of between 3 and 12 codons ending at the PTAPP motif of p6. When maternal p6 duplication was present and the p6 sequence was available from both maternal and infant isolates, all (seven of seven) infants had p6 duplications, but two cases showed discordancies between maternal and infant sequences. The prevalence of p6 duplication varied among geographical sites, ranging from 4 of 43 families (9%, Puerto Rico and New York sites) to 16 of 60 families (27%, Massachusetts, Texas, and Illinois). The presence of p6 duplication was not associated with differences in transmission, viral load, or disease progression in the infants, but showed a trend toward association with lower maternal CD4 count. Substantial p6 variation data are generated by resistance genotyping. PTAP duplication is prevalent in this group of HIV-infected women and infants. The duplication is efficiently transmitted from mother to infant, is present at variable prevalence at different geographic sites, and shows no clear association with vertical transmission risks. PMID- 15795525 TI - Anemia and HIV in the antiretroviral era: potential significance of testosterone. AB - Anemia, the most common hematological disorder in human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS), is associated with decreased quality of life and survival. Hypogonadism is prevalent in advanced HIV disease, however, low testosterone levels have not been customarily implicated in HIV-associated anemia. This study was undertaken to determine whether there is a relationship between testosterone levels and androgen use with anemia in HIV, and to characterize other clinical correlates of HIV-associated anemia. This was a cross-sectional, observational study of 200 HIV-positive patients at a public hospital HIV clinic from July 2000 to August 2001. A written questionnaire detailed previous and current medication use, opportunistic infections, and malignancies. Hematological and virological parameters, testosterone, and erythropoietin levels were measured; CD4(+) T lymphocyte count and viral load nadir and peak levels were obtained from the computerized medical record. Anemia was defined as hemoglobin <13.5 g/dl in men and <11.6 g/dl in women. Twenty-four percent of women and 28% of men were anemic. Anemia was associated with lymphopenia (adjusted OR 4.0, 95% CI 1.36-11.80), high erythropoietin levels (adjusted OR 7.73, 95% CI 2.92-20.48), and low testosterone levels (adjusted OR 3.27, 95% CI 1.01-10.60). Anemia was negatively associated with female sex (adjusted OR 0.30, 95% CI 0.11-0.85), current antiretroviral therapy (adjusted OR 0.43, 95% CI 0.20-0.95), current androgen use (adjusted OR 0.20, 95% CI 0.05 0.84), and macrocytosis (adjusted OR 0.23, 95% CI 0.09-0.61). Low testosterone levels may have a positive association and supplemental androgens a negative association with anemia in HIV disease. PMID- 15795526 TI - SPL7013 gel as a topical microbicide for prevention of vaginal transmission of SHIV89.6P in macaques. AB - SPL7013 is a dendrimer with a polyanionic outer surface that allows multiple interactions with target sites. It potently binds and blocks HIV-1 and chimeric simian/HIV-1 viruses (SHIVs) replication in vitro. Gels containing different concentrations of SPL7013 were used as topical microbicides in female pigtailed macaques (Macaca nemestrina) to study their ability to prevent vaginal transmission of SHIV(89,6P). On virus challenge, all untreated macaques (8/8) and seven of eight macaques treated with placebo gel were infected within 2 weeks postinfection (PI) and showed high plasma viremia and dramatic CD4(+) cell decline within 4 weeks PI. In contrast, 6/6 macaques, 5/6 macaques, and 2/6 macaques treated with 5% w/w (50 mg/ml), 3% w/w (30 mg/ml), and 1% w/w (10 mg/ml) SPL7013 gels, respectively, resisted viral challenge. The results showed that animals treated with SPL7013 showed a dose-dependent resistance to virus challenge. Neither SPL7013 nor placebo gels produced any adverse effects following the single application in the study. These results showed that 3-5% w/w SPL7013 gels were effective in blocking vaginal transmission of SHIV in macaques after single gel application followed by single virus challenge. These results suggest that SPL7013 gel may be a promising anti-HIV microbicide formulation for further evaluation. PMID- 15795527 TI - Beneficial effect of highly active antiretroviral therapy on the prognosis of AIDS-related systemic non-Hodgkin lymphomas. AB - The influence of HAART on the survival of patients with AIDS-related lymphoma (ARL) was evaluated. A retrospective analysis of 73 HIV-1-infected patients with proven ARL diagnosed between 1992 and 2000 was conducted. Patients received uniformly the same chemotherapy regimen according to CD4 cell counts at NHL diagnosis:, patients with CD4 cells below or above 100 cells x 10(6)/liter received CHOP or ACVBP regimens, respectively. Event-free survival (EFS) and survival were estimated by the Kaplan-Meir method and a Cox model was used to evaluate the effect of different variables on survival. At diagnosis of ARL, the median age was 37 years and 22 patients (30%) had prior AIDS-defining events. Median CD4 cell count was 99 x 10(6)/liter. The median follow-up was 60 months. Ann Arbor stage 3-4 was noted in 60 patients (82%) and bone marrow or meningeal involvement was present in 13 (17%) and 12 (16%) patients, respectively. Two groups were identified: group 1 (n = 38) included patients who had never received HAART and group 2 (n = 35) included those who received HAART either before the diagnosis or following ARL. There was no statistical significant differences in lymphoma extensive stage, presence of B symptoms, meningeal involvement, CD4 cell count at diagnosis, prior AIDS events, or chemotherapy regimens between the two groups. Median survival (MS) of the whole cohort of patients was 8 months. Estimated EFS was significantly higher (30 months) in group 2 compared to group 1 (6.1 months) (p = 0.03). In the multivariate Cox model HAART has an independent significant effect on EFS (p = 0.0085). No influence on outcome was found for other variables except for prior AIDS and bone marrow involvement. HAART has significantly improved the survival and EFS in patients with ARL, independently of chemotherapy regimen. PMID- 15795528 TI - Impact of HIV type 1 genetic subtype on the outcome of antiretroviral therapy. AB - The objective of this study was to investigate the short-term virological outcome of antiretroviral combination therapy (ART) in relation to infection with different HIV-1 genetic subtypes. Antiretroviral drug-naive patients in Sweden were prospectively enrolled and followed for 6 months when starting ART in the period from January 1998 to January 2002. Plasma-HIV-1 RNA levels, CD4 counts, and type of ART regimen were recorded. The HIV-1 subtype was determined by direct sequencing of regions of the env or pol genes. Data from 172 patients who harbored subtypes A, B, C, D, G, and CRF01_AE were analyzed (32 A, 44 B, 34 C, 18 D, 5 G, and 19 CRF01_AE). Of all patients 84% had undetectable plasma HIV-1 RNA levels after 6 months of ART. Patients infected with CRF01_AE more often had undetectable HIV-1 RNA plasma levels than patients infected with subtypes A or D. However, the possibility that this difference is due to ethnicity cannot be ruled out. Of patients of African origin, 77% had undetectable viral load after 6 months of treatment, while the corresponding figures for Caucasians and Asians were 91% and 100%, respectively. Thus, we have found an overall good short-term virological outcome after the initiation of ART in a cohort of ARV-naive patients of diverse ethnic background infected with different HIV-1 genetic subtypes. In univariate analysis ethnicity, but not genetic subtype, correlated with virological response. However, the impact of ethnicity was moderate. Patients of African origin, who had the poorest outcome, showed a 77% virological response rate. PMID- 15795529 TI - IL-8 increases transmission of HIV type 1 in cervical explant tissue. AB - Interleukin-8 (IL-8) is the predominant cytokine expressed in the female genital tract of women with certain infectious/inflammatory conditions. IL-8 increased HIV-1 replication in T cells and to a greater extent in monocytes/macrophages in vitro. Physiological levels of IL-8 increased susceptibility to HIV-1 infection 5 to 8-fold in cervical explant tissues. Competitive inhibition of the IL-8 receptor CXCR2 with the small molecule inhibitor SB225002 resulted in a 45-70% decrease in cervical explant susceptibility to HIV-1 infection. PMID- 15795530 TI - Short communication: HIV type 1 phenotype, tropism, and sequence patterns: association and preference. AB - In this study, the relationship between biological phenotypes, coreceptor usages, and sequence patterns of V1V2 or V3 regions on HIV-1 envelope gp 120 was carefully analyzed based on the existing isolates in the Los Alamos National Laboratory sequence database. Obviously, SI/NSI phenotypes were closely linked to the capability of HIV-1 to use coreceptor CXCR4, but not CCR5. Moreover, compared to NSI or R5 isolates, SI or X4 HIV-1 isolates significantly prefer higher net charges and the loss of the N-linked glycosylation site in the V3 loop, but they show no preference in either net charge or N-linked glycosylation of the V1V2 region. In addition, no significant relationship between V1V2 length and virus tropism or phenotypes was observed. PMID- 15795531 TI - Immunodominance and cross-reactivity of B5703-restricted CD8 T lymphocytes from HIV type 1 subtype C-infected Ethiopians. AB - The HLA-B57 allele family is associated with slow progression to disease in HIV-1 infected individuals and restricts a potent CD8 response against the p24 protein. This study was designed to assess the sequence variation and the CD8 response against B57-restricted epitopes of the p24 protein in a cohort of HIV-1 subtype C infected individuals possessing a high frequency of the B5703 allele. Gag sequences were amplified by PCR, cloned, and sequenced from 19 individuals including 8 B57-negative individuals. CD8 responses were assessed by interferon gamma ELISPOT assay directly from PBMC using synthetic peptides matching the autologous virus as well as the peptides representing the sequence variants circulating within the B5703 individuals. The KF11 epitope (p24 amino acids 162 172) and variants of this epitope were immunodominant in subjects possessing the B5703 allele. Three variants were observed only in B5703 individuals. Differing patterns of cross-reactivity against variant peptides were observed and were dependent upon the sequence of the autologous virus. Subjects infected with the A2G, S4N variant of KF11 demonstrated poor cross-reactivity against all other variant peptides. Determination of the breadth of viral quasispecies circulating in a population provided crucial information for studying potential escape variants of an immunodominant epitope. The presented data show that the sequence of autologous virus is critical in determining the extent of cross-reactivity of a CD8 T cell response against heterologous virus variants. Efforts to optimize the cross-reactivity of vaccine-induced CD8 T cells may need to focus on the relative immunogenicity of minor sequence variation. PMID- 15795532 TI - Short communication: pitfalls of culturing C8166 cells in serum-free media. AB - Long-term culture of C8166 cells in serum-free media can result in changes in their level of expression of immunologically important cell surface makers and a loss of infectivity by HIV-1. We have now demonstrated that these phenotypic changes are due to an outgrowth of a very small number of contaminating cells of mouse origin. Our observations emphasize the importance of carefully recharacterizing any cells that have been adapted to grow in a serum-free culture media. PMID- 15795533 TI - Using thrombelastography to determine the efficacy of the platelet glycoprotein IIb/IIIa antagonist, roxifiban, on platelet/fibrin-mediated clot dynamics in humans. AB - The effect of platelet glycoprotein IIb/IIIa antagonists on the dynamics of platelet/fibrin clot formation and strength was determined using thrombelastography (TEG) under conditions of recalcification or tissue factor addition. In the present investigation, the effect of roxifiban (class I) on ex vivo clot dynamics using recalcified blood was tested in normal, healthy volunteers (n = 7) dosed with 1 mg BID roxifiban for 9 days. Roxifiban inhibited platelet aggregation induced by 20 mumol/l adenosine diphosphate by 60-90% but did not significantly affect any of the TEG parameters either at peak, trough, or subtrough drug levels. Addition of 30 nmol/l roxifiban free acid (XV459; which is ineffective by itself to modify TEG parameters) to human blood obtained from roxifiban-treated subjects resulted in 45-60% (P < 0.01) inhibition of clot strength (maximum amplitude), 90-100% (P < 0.01) inhibition of initial kinetic of clot development (angle alpha), and 50-70% (P < 0.01) inhibition of early clot initiation (K). These data suggest that a subthreshold blood level of 40-50 nmol/l roxifiban active form was achieved in those subjects, as estimated from an in vitro calibration with XV459. These data indicate (not studied) that roxifiban, at a targeted clinical dosing regimen, failed to achieve sufficient exposure to modulate platelet-mediated clot retraction. PMID- 15795535 TI - The effect of pre-eclampsia on the levels of coagulation and fibrinolysis factors in umbilical cord blood of newborns. AB - The effect of pre-eclampsia on coagulation and fibrinolysis in newborns is still under investigation. We have evaluated several coagulation and fibrinolysis parameters in umbilical cord blood of 20 newborns from pre-eclamptic women and of 40 newborns from normotensive women with similar gestational age. Additionally, the presence of factor V Leiden and prothrombin G20210A mutation in cord blood has been assessed. Neonates from pre-eclamptic women exhibited significantly lower birth weight (2.48 +/- 0.92 versus 2.88 +/- 0.68 kg, P < 0.05) and were more frequently admitted to the neonatal intensive care unit (45 versus 20%, P < 0.01) as compared with neonates from normotensive women. Cord blood protein C antigen and activated protein C resistance mean levels were slightly higher in the group of neonates from pre-eclamptic mothers. Fibrinogen levels were lower in this group as compared with control newborns (132.17 +/- 46.97 versus 156.08 +/- 49.58 mg%, P < 0.02), and unrelated to birth weight. No significant differences between cases and controls were found in plasminogen activator inhibitor-1 or tissue plasminogen activator cord blood levels. Heterozygous prothrombin 20210A was found in three newborns from normotensive mothers, whereas no factor V Leiden mutation was found in either group. In conclusion, pre-eclampsia seems to have only mild effects on coagulation and fibrinolytic factors in the cord blood of newborns. Since no excess of common polymorphisms predisposing to thrombosis was found in newborns from pre-eclamptic mothers, it is unlikely that the carriership status of these genetic defects of newborns influences the adverse pregnancy/neonatal outcomes. PMID- 15795534 TI - Effects of pre-analytical variables on the anti-activated factor X chromogenic assay when monitoring unfractionated heparin and low molecular weight heparin anticoagulation. AB - The purpose of this study was to determine whether the anti-activated factor X (anti-FXa) assay is less affected by pre-analytical variables in monitoring patients on unfractionated heparin (UFH) and low molecular weight heparin (LMWH) than the activated partial thromboplastin time (aPTT). Forty-six subjects receiving either enoxaparin (LMWH) or UFH were randomly selected. Each study subject had six vacutainer tubes (3.8% sodium citrate, 3.2% sodium citrate) drawn by an atraumatic venipuncture. One tube from each set had a blood to anticoagulant ratio of 9: 1. The other tube had an intentional "short-draw" of approximately 6: 1 blood to anticoagulant ratio. All specimens had an aPTT and a chromogenic anti-FXa assay performed on each specimen regardless of heparin type. The aPTT assay mean with the 3.8% sodium citrate tube short-draw tube was statistically different from the other aPTT assays (P = 0.06). However, all six of the mean anti-FXa assays for the UFH and LMWH heparin subjects were not statistically or clinically different (analysis of variance, P = 0.9878 for UFH and P = 0.9060 for LMWH). The intentional short-draw tube did not affect the anti FXa assay regardless of the anticoagulant. The anti-FXa assay appears to be a better method for monitoring heparin subjects than the aPTT due to the lack of effect of pre-analytical variables. PMID- 15795537 TI - Study of human von Willebrand factor immunogenicity in pigs with severe von Willebrand disease. AB - Replacement therapy is the treatment of choice for patients with von Willebrand disease who are unresponsive to desmopressin. In order to prevent transmission of non-enveloped viruses, a solvent/detergent-treated plasma-derived von Willebrand factor available in France since 1989 has been subjected to additional removal/inactivation steps by 35 nm filtration and dry heating for 72 h at 80 degrees C. This preclinical study evaluates the potential immunogenicity of this new product by comparing the antibodies raised in pigs affected with von Willebrand disease after intravenous injection of either a solvent/detergent treated product or a triple-secured product. Our data showed that there is no difference between the two products in terms of the rate and intensity of the humoral response measured by both binding and neutralizing antibody levels. It was concluded that no antigenic alterations of von Willebrand factor molecules during the nanofiltration and final dry-heating steps were detected in our animal model. PMID- 15795536 TI - Mechanical heart valve prostheses and persistent lupus anticoagulant: is the thrombotic risk increased? AB - The risk of thrombosis in patients with mechanical heart valve prostheses in spite of life-long adequate anticoagulation is 1-2% per year. Current recommendations for anticoagulation take into account the prosthesis itself and the co-morbid conditions that enhance the thrombotic risk. Lupus anticoagulant is diagnosed in many thrombotic recurrences. We designed an ambispective case control study to evaluate thrombotic events in patients with mechanical heart valve prostheses and persistent lupus anticoagulant. Our objectives were to determine whether persistent lupus anticoagulant increased the risk of embolism in that population and thus, if a more intense anticoagulation would be recommended, even at the risk of increasing bleeding episodes. We included 16 patients and 16 controls with more than 80 patient-years of follow-up and with other risk factors for embolism. We observed no increased rate of thromboembolic events in patients than in controls, even during high-risk situations (i.e. bacterial endocarditis). Our population spent most of the time within the intended anticoagulation range. We conclude that adequate anticoagulation is the most important issue to prevent events, protecting against thrombosis without increasing the bleeding risk. PMID- 15795538 TI - No association between pulmonary embolism or deep vein thrombosis and the -455G/A beta-fibrinogen gene polymorphism. AB - Hyperfibrinogenaemia has been reported to be associated with deep vein thrombosis (DVT). However, whether or not the "fibrinogen-raising"-455G/A polymorphism of the beta-fibrinogen gene is associated with DVT is uncertain and there are no data on whether this polymorphism is associated with pulmonary embolism (PE). We have studied relationships between the -455G/A beta-fibrinogen gene polymorphism and the occurrence of PE and/or DVT (n = 339) (PE only, n = 76; DVT only, n = 216; PE and DVT, n = 47). There was no difference between the -455A allelic frequencies for the control (n = 190) and patient groups - PE, 0.187 and 0.171, respectively [P = 0.6087, chi test; odds ratio (OR), 1.12; 95% confidence interval (CI), 0.72-1.74]; DVT, 0.187 and 0.171, respectively (P = 0.5408, chi test; OR, 1.11; 95% CI, 0.78-1.59). This also applied when only Caucasian individuals were considered - PE allelic frequencies, 0.192 and 0.193, respectively (P = 0.9764, chi test; OR, 0.99; 95% CI, 0.62-1.60); DVT allelic frequencies, 0.192 and 0.186, respectively (P = 0.8404, chi test; OR, 1.04; 95% CI, 0.71-1.51). While the results should be interpreted with caution as the frequency of the -455A allele is rare, the -455A allele of the beta-fibrinogen gene does not appear to be associated with an increased risk of PE or DVT. PMID- 15795539 TI - P2Y12 gene H2 haplotype is not associated with increased adenosine diphosphate induced platelet aggregation after initiation of clopidogrel therapy with a high loading dose. AB - A large variability in the antiplatelet response to clopidogrel has been consistently reported. Recently, a P2Y12 haplotype was shown to be associated with enhanced adenosine diphosphate (ADP)-induced platelet aggregation in healthy volunteers. The aim of this study was to test in patients (n = 416) scheduled for coronary artery stenting whether P2Y12 haplotype H2 carriage is associated with increased ADP-induced platelet aggregation after administration of a 600 mg loading dose of clopidogrel. Blood was drawn from the arterial sheath at least 2 h after administration of 100 mg aspirin and 600 mg clopidogrel. ADP-induced platelet aggregation was assessed in platelet-rich plasma with light-transmission aggregometry. P2Y12 haplotypes (H1/H2) and P2Y12 C32T genotypes were determined with TaqMan assays. Haplotype combinations and genotypes were not associated with parameters of ADP-induced platelet aggregation after administration of a 600 mg loading dose of clopidogrel. Maximal ADP (5 mumol/l)-induced platelet aggregation was similar in patients carrying haplotype H2 and homozygous carriers of haplotype H1 (43.9 +/- 21.4 versus 43.2 +/- 21.1%, respectively; P = 0.77). Carriage of P2Y12 H2 haplotype does not seem to affect the platelet response to a 600 mg loading dose of clopidogrel in coronary artery disease patients prior to stenting. PMID- 15795540 TI - gammaAla82Gly represents a common fibrinogen gamma-chain variant in Caucasians. AB - Screening of 200 blood donors for the presence of polymorphisms in three fibrinogen genes (FGA, FGB, FGG), revealed two individuals with a heterozygous missense mutation (c.323C > G, gammaAla82Gly) in the FGG gene. This mutation has been reported previously to cause mild hypofibrinogenaemia. Analysis of an additional 416 blood donors showed two more heterozygous gammaAla82Gly mutations, resulting in an overall gammaAla82Gly allele frequency of 0.0032. Haplotype analysis demonstrated that the gammaAla82Gly mutation originated from a common founder. From these data we estimated that homozygous individuals for gammaAla82Gly should occur at a frequency of 1: 95 000, suggesting that hypofibrinogenaemia represents a more frequent condition in the population than so far believed. PMID- 15795541 TI - Plasma levels of tissue factor and soluble E-selectin in sickle cell disease: relationship to genotype and to inflammation. AB - BACKGROUND: Microvascular occlusion, the pathophysiological hallmark of sickle cell disease (SCD), is a complex multifactorial process with alterations in coagulation, endothelial function and inflammation. However, relationships between these process in the two most common genotypes, HbSS and HbSC, are unknown. We hypothesized differences in the hypercoagulable state [as assessed by tissue factor (TF), fibrinogen and D-dimer], endothelial function [markers soluble E-selectin (sE-sel) and von Willebrand factor (vWf)], and inflammation [markers interleukin-6 (IL-6) and high-sensitivity C-reactive protein (hsCRP)] in these two SCD genotypes. Citrated plasma TF, sE-sel, vWf, fibrinogen and fibrin D dimer, and serum IL-6 and hsCRP (enzyme-linked immunosorbent assay/Clauss) were measured in 64 patients with SCD (27 with HbSS disease) and 42 AA subjects matched for age and ethnic origin. TF (P = 0.0014), sE-sel (P = 0.001) and, as expected, vWf, D-dimer, and hsCRP (all P < or = 0.01), but not fibrinogen or IL 6, were raised in the SCD patients compared with the AA subjects. However, only vWf and, as expected, D-dimer (all P < or = 0.01) were higher in HbSS disease than in HbSC disease. Raised plasma TF and sE-sel in SCD compared with HbAA subjects may contribute to the increased risk of thrombotic disease in this group. Raised vWf in HbSS compared with HbSC may be important in determining pathophysiology in these two genotypes. Positive correlations between IL-6 and TF in both HbSC and HbSS disease leads us to speculate that inflammation may be important in coagulation activation in these patients, or vice versa. However, lack of correlation of sE-sel with inflammatory markers implies that other mechanisms are responsible for increased levels of this marker of endothelial activation. PMID- 15795542 TI - Activated protein C resistance acquired through liver transplantation. AB - Coagulation factors are produced by the liver. It is well recognized that liver transplantation can cure haemophilia. We described a case of thrombophilia acquired due to liver transplantation from a donor with heritable thrombophilia. PMID- 15795543 TI - Prophylaxis with enoxaparin can produce a giant abdominal wall haematoma when associated with low doses of aspirin among elderly patients suffering cough attacks. AB - We report three patients aged 75-80 years observed in the Emergency Room with severe anaemia (requiring transfusion) due to a large abdominal wall haematoma while receiving standard prophylaxis against venous thromboembolism (40 mg/day enoxaparin for 6 days on average). All of them concomitantly received 100 mg/day aspirin because of previous ischaemic heart disease and presented similar clinical features: sudden onset of abdominal pain during a severe cough episode due to bronchial infection. A giant haematoma in the rectus abdominis muscle was recognized (by computed tomography) in every case. The cough has been related with this complication in some reports but its association with antiplatelet drugs and low molecular weight heparin could increase the risks in older patients. Sudden abdominal pain must alert the clinician to the rectus muscle sheath haematoma in order to avoid the risks of an exploratory laparotomy. PMID- 15795544 TI - A novel nonsense mutation in the FGA gene in a Chinese family with congenital afibrinogenaemia. AB - Congenital afibrinogenaemia is a rare autosomal recessive coagulation disorder. Here we describe the genetic defect in the fibrinogen A alpha-chain underlying afibrinogenaemia in a Chinese family. The proposita had a life-long bleeding tendency, both her parents and paternal grandparents had a consanguineous marriage. The blood-clotting indices of the proposita and her father were prolonged, and their functional and immunologic fibrinogen was absent. To identify the mutations of fibrinogen genes in this family, all the exons and exon intron boundaries of the three fibrinogen genes (FGA, FGB, FGG) were amplified by polymerase chain reaction, and direct sequencing of polymerase chain reaction products was performed, then the restriction endonuclease (RsaI) analysis was used to confirm the mutation. A homozygous C --> T mutation was found at nucleotide 3108 in exon 4 of the FGA gene of the proposita and her father; it is a null mutation predicting to produce severely truncated A alpha-chains because of the presence of premature termination at the Gln 150 codon (or truncated at the 131 residues according to the mature A alpha-chain). Her mother and some other family members were heterozygous. The g.3108C --> T (Gln150 --> stop) nonsense mutation in the FGA gene is a novel genetic defect of congenital afibrinogenaemia that, to our knowledge, has not been described previously. PMID- 15795545 TI - Monitoring oral anticoagulant therapy by telephone communication. AB - The number of patients who need supervision during oral anticoagulant treatment is growing constantly. We have presently enrolled 156 patients who were referred to our anticoagulant clinic and who were taking sodium warfarin with target International Normalized Ratios (INR) of low (2-3), intermediate (2.5-3.5) and high (3-4) range. Patients performed the tests in laboratories situated in locations at their convenience and received further instructions from a specialist via telephone communication. A total of 8758 prothrombin times (5214, 1947 and 1597 tests for individuals in the low, intermediate and high range, respectively) were performed over the period of 3.16 +/- 2.6 years (range, 6 months-9.5 years) and reported to the specialist. It was found that in the aforementioned three groups of intensity 63.3, 57.0 and 47.7% of the INRs were within the target range, the respective percentages for the expanded (+/- 0.5) target INR being 92.8, 87.8 and 78.5%. The INTERDAY software was used to calculate the number and proportion of days within the target INR range, the respective results being 71.0, 64.0 and 51.6% and 96.2, 93.2, 86.4% for the expanded range. The number and percentage of bleeding and embolic complications' referrals to the emergency room and hospitalizations were similar to those reported for anticoagulant clinics in which patients have to actually pay a personal visit in order to receive instructions. Our study is significant in that it documents that trans-telephonic communication is feasible safe and cost effective and that the clinical results are at least as good as those obtained by traditional consultation. PMID- 15795546 TI - The significance of prestroke aspirin dosage in fatal outcome of acute stroke. AB - BACKGROUND: Aspirin is an effective and generally accepted treatment drug during the acute stage of ischemic brain infarction. The association between the pretreatment aspirin dosage and fatal outcome among these treated patient groups has not been analyzed previously. OBJECTIVE: The aim of the study was to evaluate 14- and 30-day poststroke survival in patients treated with 325 mg immediately on admission to the hospital for prestroke antiaggregation/anticoagulation treatment. METHODS: This prospective study was conducted in a government tertiary care facility. The medical records of all 1245 patients admitted to the hospital for stroke from 1997 to 2002 were reviewed. The association between demographic parameters, stroke risk factors, stroke subtype, prestroke antiaggregation/anticoagulation treatment, and risk of fatal poststroke outcome, 14 days and 30 days after the event, was analyzed using Cox survival analyses. RESULTS: During the 14-day poststroke period, 320 patients (25%) died. By day 30 poststroke, 386 patients (31%) had died. Older age, female gender, chronic heart disease, and cardiac arrhythmias were associated with increased risk of increased fatal outcome. Mortality was higher in patients with stroke caused by cardioembolism (P < 0.0001) and was significantly lower in patients with small vessel occlusion (P < 0.0001). Prestroke medium-dose aspirin treatment was associated with a relative reduction in 30-day poststroke period mortality (P < 0.0001). Conversely, prestroke low-dose aspirin treatment was associated with increased mortality (P < 0.0001). CONCLUSION: Prestroke medium-dose aspirin treatment was associated with reduced 30-day poststroke mortality, whereas low dose prestroke aspirin therapy was associated with increased 30-day poststroke mortality. PMID- 15795548 TI - Augmentation with amisulpride for schizophrenic patients nonresponsive to antipsychotic monotherapy. AB - Despite the effectiveness of antipsychotic medications in treatment of schizophrenia, about 30% of patients who receive an adequate treatment have significant persisting symptoms. The problem of treatment-resistant psychosis is an important and difficult one. The aim of this study was to retrospectively evaluate the efficacy and safety of amisulpride augmentation in treatment resistant schizophrenic patients. To the best of our knowledge, this is the first report about resistant schizophrenic and schizoaffective patients treated with the combinations of risperidone and amisulpride and ziprasidone and amisulpride. Data were collected from patient records. A total of 15 resistant schizophrenic patients (7 men, 8 women, 54.0 +/- 16.9 years old) were included in the study. Before addition of amisulpride, the patients were treated with monotherapy by atypical neuroleptics (clozapine, olanzapine, risperidone, or ziprasidone). The mean amisulpride dose was 693.3 +/- 279.6 mg/d. The mental state of 12 (80%) patients treated with combination was improved. Three (20%) patients showed no change in their mental state. Only 2 patients treated with a combination of risperidone and amisulpride had mild side effects. The results are preliminary and require confirmation in a randomized controlled trial. The authors suggest that amisulpride may be a promising option as an augmentation strategy in treatment-resistant schizophrenic patients. PMID- 15795547 TI - Aggravation of absence seizures by carbamazepine in a genetic rat model does not induce neuronal c-Fos activation. AB - The mechanisms underlying carbamazepine aggravation of absence seizures are uncertain but are thought to involve enhancement of neuronal activity within the thalamocortical circuitry. We used c-Fos immunohistochemistry (cFos-ir) to examine patterns of neuronal activation and the relationship to seizure expression following administration of carbamazepine in a rat model of absence epilepsy (Genetic Absence Epilepsy Rats of Strasbourg, GAERS). Female ovariectomized GAERS implanted with extradural EEG electrodes received either 20 mg/kg carbamazepine or vehicle IP. Seizure expression was quantified by measuring the total number and duration of spike-wave discharges (SWD) and with the individual burst discharge lengths over a 90-minute EEG. This was correlated with cFos-ir in thalamocortical slices from rats killed 180 minutes after carbamazepine administration. Carbamazepine-treated rats (n = 5) had a significantly greater total duration of SWD than vehicle-treated rats (17.9% versus 8.8%, P = 0.04). Despite this aggravation of seizures, the level of cFos ir did not differ between the treatment groups. A positive correlation was found between cFos-ir in the reticularis thalami (Rt) and the total seizure duration (R = 0.66, P = 0.04) and mean burst length (R = 0.68, P = 0.03) but not total number of seizures. The lack of difference in cFos activation patterns between carbamazepine and vehicle-treated animals suggests that the mechanism for carbamazepine aggravation of absence seizures may not involve neuronal activation but rather enhanced neuronal synchronization. The association between increased neuronal activation in the Rt and seizure burden in GAERS provides further support for the critical role of this structure in the maintenance, but not initiation, of absence seizure activity. PMID- 15795549 TI - Meta-analysis and indirect comparisons of levetiracetam with other second generation antiepileptic drugs in partial epilepsy. AB - Few comparative clinical trials of newer antiepileptic drugs (AEDs) in patients with refractory partial epilepsy are available. Therefore, meta-analysis is a widely used and useful method for comparing them. Despite the limitations of indirect comparisons, and recognizing that these drugs were tested at different doses, such comparisons can be helpful to physicians making practical treatment decisions. The purposes of this study were to present newer meta-analysis results for add-on levetiracetam compared with placebo and to estimate its efficacy and tolerability compared with other new AEDs (gabapentin, lamotrigine, oxcarbazepine, tiagabine, topiramate, and zonisamide) in a meta-analysis using methods for making indirect comparisons. Randomized placebo-controlled clinical trials of add-on therapy with levetiracetam, gabapentin, lamotrigine, oxcarbazepine, tiagabine, topiramate, and zonisamide in patients with refractory partial epilepsy were identified in the Cochrane Library 2002. A fixed-effects model was used to estimate Mantel-Haenszel odds ratios for the responder rate (efficacy measure) and withdrawal rate (mainly tolerability measure) of levetiracetam and other new AEDs versus placebo. Because no head-to-head clinical trials comparing these new AEDs exist, adjusted indirect comparisons were then made between levetiracetam and each other AED using the meta-analysis results. At the doses tested, levetiracetam was more effective in terms of responder rate than gabapentin (odds ratio 2.64 with 95% CI 1.51-4.63) and lamotrigine (odds ratio 1.86 with 95% CI 1.04-3.34) and equally well tolerated. Levetiracetam had a significantly lower withdrawal rate than topiramate (odds ratio 0.52 with 95% CI 0.29-0.93) and oxcarbazepine (odds ratio 0.55 with 95% CI 0.33-0.92), with comparable efficacy. Although levetiracetam did not differ significantly from the other AEDs, numerical trends favoring levetiracetam were obtained in response rate and in withdrawal rate (tiagabine, zonisamide). Indirect comparisons based on meta-analysis suggest that add-on therapy with levetiracetam has a favorable responder and/or withdrawal rate relative to several AEDs in patients with partial epilepsy with doses used in clinical trials. These meta-analyses give only short-term efficacy and safety data. Comparative clinical trials and long term studies of these agents are needed to confirm these findings. PMID- 15795550 TI - The relation between visual hallucinations and visual evoked potential in Parkinson disease. AB - OBJECTIVE: The pathophysiology of hallucinations in Parkinson disease is poorly understood. This study investigated the relation between visual hallucination and visual evoked potentials (VEPs) in Parkinson disease. METHODS: Nineteen patients with Parkinson disease were studied. The authors divided patients into 2 groups: patients with visual hallucinations (VH group) and those without visual hallucinations (no-VH group). VEPs using a checkerboard stimulus were recorded under a drug-free state. RESULTS: On multivariate regression analysis, only the average P100 latency was selected and remained significant after the backward elimination method. CONCLUSION: The authors demonstrated a close association between visual hallucinations and elongated VEP latency in Parkinson disease. VEPs may become one of the predictors for visual hallucination. PMID- 15795552 TI - Suppression of craving for gamma-hydroxybutyric acid by naltrexone administration: three case reports. AB - Gamma-hydroxybutyric acid (GHB) is currently used to induce and maintain abstinence from alcohol. Cases of craving and desire to increase doses of GHB have been reported in both clinical trials and nonclinical self-administration. The enhancement of dopamine activity induced by GHB receptor activation might play a role in the euphoric effect and potential craving and the consequent abuse of this drug. Naltrexone (NTX), a mu-opioid antagonist, is effective in inducing and maintaining abstinence from alcohol, reducing relapses in heavy drinking and craving for alcohol in alcohol-dependent outpatients. Taking into account the alcohol antireward property of NTX, we tested its activity in reducing craving for GHB in 3 consecutive cases of alcoholics who manifested craving for this drug. In all patients the combination with NTX suppressed the craving for GHB. The antireward effect of NTX likely results from its interference with the GHB induced dopamine release, leading to a partial blockade of the GHB reinforcing effect responsible of the craving for the drug. A combined therapy with GHB and NTX seems to be able to suppress craving for the former, thus improving the manageability and safety of treatment. PMID- 15795551 TI - Ziprasidone-associated mania: a review and report of 2 additional cases. AB - Although mania was not reported as an adverse event in the pivotal trials of ziprasidone, there have been 7 reports of ziprasidone-induced mania in 12 patients. We now report 2 additional cases wherein the introduction of ziprasidone resulted in new-onset manic episodes. In 1 case, the patient required hospitalization and lost his job. In the other, time to mania was 5 months, considerably longer than previously reported. Of the 14 cases, 9 were in a depressive episode when ziprasidone was prescribed, and 8 had a history of current or past exposure to antidepressants. Ziprasidone, like many antidepressants, can block reuptake of norepinephrine and serotonin, although mania has developed in patients treated with atypical antipsychotics that are less potent in this regard. However, ziprasidone and other atypical antipsychotics have in common a high ratio of 5-HT2a to D2 receptor blockade, which may also play a role in this phenomenon. Clinicians and patients need to be aware of the potential for the induction of mania with ziprasidone, even after lengthy exposure. PMID- 15795554 TI - Severe topiramate-associated hyperthermia resulting in persistent neurological dysfunction. AB - Topiramate has recently been reported to cause hyperthermia as a result of oligohydrosis, primarily in pediatric patients. All cases reported to date were clinically mild, without permanent systemic or neurologic dysfunction. We report a case of severe hyperthermia and subsequent ataxia and tremor in an adult treated with topiramate. To our knowledge, this is the first case of topiramate associated hyperthermia to result in residual cerebellar and cognitive dysfunction. PMID- 15795553 TI - Risperidone withdrawal-related respiratory dyskinesia: a case diagnosed by spirography and fibroscopy. AB - Risperidone is a frequently used member of a new class of atypical antipsychotics the serotonin-dopamine antagonists (SDAs)-due to its comparatively high efficacy and low D2/5HT2 binding ratio, which results in a low incidence of extrapyramidal side effects including tardive dyskinesia (TD). The authors present an elderly patient with mixed dementia who developed TD at multiple sites, (including respiratory dyskinesia [RD], limb dyskinesia, and orofacial dyskinesia) following abrupt withdrawal of risperidone therapy. RD is not a rare condition, but is often misdiagnosed and is potentially lethal. Therefore, clinicians should pay close attention to possible onset of RD in patients with multiple risk factors for TD, even when SDA therapy is used. If RD is suspected, assessment should include spirography combined with fibroscopy for examination of irregularities in the rate, rhythm, and depth of respiration. PMID- 15795555 TI - Psychotropic drug interactions with valproate. AB - Valproate is a well-established anticonvulsant that is increasingly being employed, often in combination with other psychotropics, for its mood-stabilizing properties. This compound is metabolized by conjugation, beta-oxidation, and cytochrome P450 oxidation (CYP2C9, CYP2C19, and CYP2A6) and also acts as a broad spectrum inhibitor of a variety of hepatic enzymes including glucoronyltransferase, epoxide hydrolase, and the CYP2C enzymes. In addition, it exhibits saturable protein binding and competes with many drugs for protein binding sites. It is therefore not surprising that valproate has been noted to interact with psychotropic medications of all classes. This article provides an overview of the noted pharmacokinetic psychotropic interactions with valproate, with a particular focus on the mechanisms of these interactions and potential clinical consequences. PMID- 15795556 TI - Levetiracetam in L-dopa-induced dyskinesia. PMID- 15795560 TI - Angiotensin-converting enzyme inhibitors, AG receptor blockers, and aldosterone receptor antagonists. PMID- 15795559 TI - Nuclear peroxisome proliferator-activated receptors and thiazolidinediones. PMID- 15795561 TI - Direct thrombin inhibitors. PMID- 15795562 TI - Statins and cardiovascular risks. PMID- 15795563 TI - Na+-K+ exchange inhibitors. PMID- 15795564 TI - Caspase inhibitors and myocardial apoptosis. PMID- 15795566 TI - Inhaled selective pulmonary vasodilators. PMID- 15795565 TI - Intracoronary brachytherapy and drug-eluting coronary stents. PMID- 15795567 TI - Calcium sensitizers, a new class of inodilators. PMID- 15795568 TI - Tifacogin, recombinant tissue factor pathway inhibitor. PMID- 15795570 TI - Trochanteric gamma nail and compression hip screw for trochanteric fractures: a randomized, prospective, comparative study in 210 elderly patients with a new design of the gamma nail. AB - OBJECTIVES: To compare the results between a new intramedullary Gamma nail and a compression hip screw in the treatment of trochanteric fractures. DESIGN: Prospective randomized. SETTING: Level 1 trauma center. PATIENTS: Two hundred ten consecutive patients older than 65 years with trochanteric femoral fractures. INTERVENTIONS: Treatment with a compression hip screw or a new design of the Trochanteric Gamma nail (180 mm in length with a mediolateral angle of 4 degrees and available only with a proximal diameter of 17 mm and distal diameter of 11 mm). MAIN OUTCOME MEASUREMENTS: Operative and fluoroscopy times, blood loss, functional outcome, complication rate, and failure of fixation. RESULTS: The Trochanteric Gamma nail was used in 104 patients and the compression hip screw in 106. The 2 groups were similar in terms of their preoperative data, with a median follow-up of 13.6 months (range 12-30). The results show no difference in operating time (P = 0.21), but the Trochanteric Gamma nail group had a significantly shorter fluoroscopy time (P = 0.006), and the number of patients transfused and the mean of units of blood transfused were significantly less in the Trochanteric Gamma nail group (P = 0.013, 0.046, respectively). Mortality within 12 months was similar in both groups (P = 0.83). All fractures were radiographically healed at the last visit. There was no difference in intraoperative and postoperative complications or rate of fixation failure between the 2 groups, and no case of secondary shaft fracture of the femur was encountered in this study. In the entire series, there was no difference in the functional outcome (P = 0.74), but the postoperative walking ability was better in those patients with unstable fractures who were treated with the Trochanteric Gamma nail (P = 0.017). CONCLUSIONS: The new Trochanteric Gamma nail is an effective method for the treatment of trochanteric femoral fractures in elderly patients. The indication for either Trochanteric Gamma nail or compression hip screw is similar in stable fractures, but we recommend the use of the Trochanteric Gamma nail for unstable trochanteric fractures. PMID- 15795571 TI - Thromboembolic disease prophylaxis in patients with hip fracture: a multimodal approach. AB - OBJECTIVES: To assess if pneumatic compression in conjunction with chemoprophylaxis is an effective way to reduce the incidence of deep vein thrombosis in orthopedic trauma patients sustaining fragility hip fractures. DESIGN: Two hundred patients admitted to the authors' institution between May 1998 and June 2002 for fractures of the hip were prospectively studied. All patients were treated operatively and received the VenaFlow calf compression device on both lower extremities immediately following surgery. Chemical prophylaxis of either aspirin (n = 67) or warfarin (n = 133) was administered in addition to mechanical compression. A noninvasive serial color flow duplex scan was performed 1 to 11 days postoperatively (mean 4.5 days) to determine the presence or absence of deep vein thrombosis. All patients were followed clinically 3 months postoperatively for a clinical evaluation of symptomatic deep vein thrombosis or pulmonary embolism. RESULTS: Overall, the incidence of deep vein thrombosis was 3.5% (7 of 200) and included only 1 proximal thrombosis (1 out of 200, or 0.5%) and no pulmonary embolism. Five of the 7 patients positive for deep vein thrombosis were in the mechanical compression and warfarin prophylaxis group and 2 were in the aspirin arm of the study. For patients with deep vein thrombosis, the average number of risk factors was 3.71, whereas patients without clots averaged 1.75 clinical risk factors (P < or = 0.05). Three patients in the warfarin group developed bleeding complications (1 with a gastrointestinal bleed and 2 with minor bleeding not at the operative site). No evidence of a symptomatic deep vein thrombosis or pulmonary embolism was reported within a 3-month period following hospitalization. CONCLUSIONS: Our findings suggest mechanical compression with the VenaFlow calf compression device in conjunction with chemoprophylaxis is an effective means of reducing thromboembolic disease in this high-risk population. PMID- 15795573 TI - Functional outcome of bilateral limb threatening: lower extremity injuries at two years postinjury. AB - OBJECTIVES: To describe the functional outcome of bilateral limb-threatening injuries at 2 years postinjury and to evaluate whether a different decision making process should be used for these patients as opposed to patients with unilateral limb-threatening injury. DESIGN/SETTING/PATIENTS: This study population of 32 patients is a subset of 601 patients from a study of 8 level I trauma centers. The patients were prospectively followed through 24 months. MAIN OUTCOME MEASUREMENTS: The principle outcome measure at 2 years was the Sickness Impact Profile, designed to measure physical and psychosocial dimensions. RESULTS: The overall Sickness Impact Profile scores at 2 years demonstrate that all 3 bilateral injury groups (bilateral salvage [n = 14], unilateral salvage/amputation [n = 8], and bilateral amputation [n = 10]) were severely disabled (Sickness Impact Profile >10). The bilateral salvage group had the most dramatic improvement over the 24 months. The 2-year physical subscale Sickness Impact Profile data showed a similar trend. At the 2-year assessment, the bilateral amputation group was recording greater disability (Sickness Impact Profile = 16.3) compared to the bilateral salvage and unilateral amputation/salvage groups (Sickness Impact Profile = 8.5 and 12.6, respectively). The overall Psychosocial Dimension, which started off worst in the bilateral salvage group, ended up similar in all 3 groups (8 to 9). The percent of patients who returned to work was 66.7% in the unilateral salvage/amputation group versus 21.4 and 16% in the bilateral salvage and amputation groups, respectively. CONCLUSIONS: The results indicate that treatment judgments should be based upon the results derived from the analysis of the larger unilateral limb cohort data. Patients with severe, bilateral lower extremity injuries should be counseled that regardless of treatment combinations, the function of each limb is similar at 24 months. The unilateral amputation/salvage group had a greater probability of going back to work. This is the major identifiable benefit to undergoing salvage versus amputation. PMID- 15795572 TI - Minimal internal fixation augmented by small wire transfixion frames for high energy tibial plateau fractures. AB - PURPOSE: To evaluate the outcome of bicondylar tibial plateau fractures treated with minimal internal fixation augmented by small wire external fixation frames and to assess the necessity of bridging the knee joint by extending the external fixation to the distal femur. METHODS: This is a retrospective study of 48 tibial plateau fractures. There were 40 (83.5%) Schatzker type VI fractures, 8 Schatzker type V fractures, and 18 (37.5%) fractures were open. A complex injury according to the Tscherne-Lobenhoffer classification was recorded in 30 (62.5%) patients. All fractures were treated with combined minimally invasive internal and external fixation. Closed reduction was achieved in 32 (66.6%) of the fractures. Extension of the external fixation to the distal femur was done in 30 (62.5%) fractures. Results were assessed according to the criteria of Honkonen-Jarvinen. RESULTS: Follow-up ranged from 28 to 60 months with an average of 38 months. All fractures but 1 united at an average of 13.5 weeks (range 11-18 weeks). One patient developed an infected nonunion of the diaphyseal segment of his fracture. Thirty nine (81%) patients achieved an excellent or good radiologic result. An excellent or good final clinical result was recorded in 36 patients (76%). Bridging the knee joint did not affect significantly the result (P < 0.418). No significant correlation was found between the type of fracture and the final score (P < 0.458). CONCLUSIONS: Hybrid internal and external fixation combined with tibiofemoral extension of the fixation is an attractive treatment option for complex tibial plateau fractures. PMID- 15795574 TI - Standard multiplanar fluoroscopy versus a fluoroscopically based navigation system for the percutaneous insertion of iliosacral screws: a cadaver model. AB - OBJECTIVES: To compare the safety and efficiency of standard multiplanar fluoroscopy (StdFluoro) and virtual fluoroscopy (VirtualFluoro) for use in the percutaneous insertion of iliosacral screws. DESIGN: : Human cadaver study comparing 2 imaging modalities during iliosacral screw insertion; imaging randomized from side to side. SETTING: Bioskills laboratory in a medical school. PARTICIPANTS: Twenty-nine embalmed whole human cadavers without prior hip or pelvic surgery. INTERVENTION: Iliosacral screws were inserted into the S1 bodies using a percutaneous insertion technique. Screws were inserted on one side using StdFluoro, and on the other side, screws were placed using VirtualFluoro. MAIN OUTCOME MEASUREMENTS: Time necessary for imaging preparation, screw insertion, and actual fluoroscopy were recorded. Accuracy and safety of screw placement was assessed using computed tomography and an anatomic dissection of the pelvis. RESULTS: : Fifty-six of 58 iliosacral screws were placed within the desired bony corridor of the posterior pelvis. One screw placed using each method was inserted erroneously, but both were relatively minor deviations. There were no obvious injuries to major vessels or nerve roots. The total surgical time required for preparation of imaging and screw insertion averaged 7.3 minutes using StdFluoro and 6.7 minutes using VirtualFluoro (P = 0.4). Although the time necessary for screw insertion using VirtualFluoro averaged only 3.5 minutes, compared to 7.0 minutes for StdFluoro (P < 0.05), this time savings was offset by that required for application and calibration of tracking devices when using VirtualFluoro. The average fluoroscopy time using StdFluoro method was 26 seconds, whereas that for the VirtualFluoro was only 6 seconds (P < 0.01). CONCLUSIONS: Most of the percutaneous iliosacral screws were safely inserted using StdFluoro and VirtualFluoro, and total surgical times were similar using both methods. As VirtualFluoro continues to evolve, improved efficiency in operative times may be expected. Currently, the most beneficial aspect of using VirtualFluoro during the insertion of percutaneous iliosacral screws appears to be significantly decreased use of fluoroscopy when compared to StdFluoro. PMID- 15795575 TI - Intraoperative three-dimensional imaging with a motorized mobile C-arm (SIREMOBIL ISO-C-3D) in foot and ankle trauma care: a preliminary report. AB - OBJECTIVE: The aim of the study was to assess the feasibility and benefit of the intraoperative use of a mobile C-arm with 3-dimensional imaging (ISO-C-3D). DESIGN: Prospective consecutive clinical study. SETTING: University hospital, level I trauma center. METHODS: The ISO-C-3D was used for intraoperative visualization in foot and ankle trauma care. Conventional C-arms were used to judge the reduction and implant position before the ISO-C-3D was used. Time spent, changes resulting from use of the ISO-C-3D, and surgeons' ratings (visual analogue scale, 0-10 points) were recorded. PATIENTS: Between January 1, 2003 and March 15, 2004, the ISO-C-3D was used in 62 cases (factures: pilon, n = 1; Weber C ankles, n = 7; isolated dorsal Volkmann, n = 1; talus, n = 3; calcaneus, n = 20; navicular, n = 1; cuboid, n = 1; Lisfranc fracture-dislocation, n = 6; hindfoot arthrodesis with or without correction, n = 12). RESULTS: On average, the operation was interrupted for 440 seconds (range 330-700); 120 seconds, on average, for the ISO-C-3D scan and 210 seconds, on average, for evaluation of the images by the surgeon. In 39% of the cases (24 of 62), the reduction and/or implant position was corrected during the same procedure after the ISO-C-3D scan. The ratings of the 8 surgeons who used the ISO-C-3D were 9.2(5.2-10) for feasibility, 9.5 (6.1-10) for accuracy, and 8.2 (4.5-10) for clinical benefit. CONCLUSION: Intraoperative 3-dimensional visualization with the ISO-C-3D can provide useful information in foot and ankle trauma care that cannot be obtained from plain films or conventional C-arms. During the same procedure, after conventional C-arm scans judged the positioning to be correct and an ISO-C-3D scan was done, the reduction and/or implant position was corrected in 39% of the cases in this study, although not unnecessarily prolonging the operation. The ISO C-3D appears to be most helpful in procedures with a closed reduction and internal fixation, and/or when axial reformations provide information that is not possible to obtain with a conventional C-arm and/or direct visualization during open reduction and internal fixation. PMID- 15795576 TI - Use of tissue ultrafiltration for treatment of compartment syndrome: a pilot study using porcine hindlimbs. AB - OBJECTIVES: To demonstrate the efficacy of compartment syndrome ultrafiltration for the treatment of acute compartment syndrome in an animal model. Our hypothesis is the removal of interstitial fluid will result in a reduction of intramuscular pressure compared with untreated controls in a model of bilateral induced compartment syndrome. DESIGN: Controlled experimental model. SETTING: Animal research facility. PATIENTS/PARTICIPANTS: Three pairs of porcine hindlimbs. INTERVENTION: Acute compartment syndrome was created in the pig hindlimb by infusion of saline to maintain the intramuscular pressure 30 mm Hg greater than the animal's mean arterial pressure for 8 hours. After a 2-hour reperfusion interval, ultrafiltration (removal of fluid through 1 mm diameter porous catheters, connected to -500 mm Hg suction) was commenced in 1 limb only and continued for 9.5 hours. MAIN OUTCOME MEASURES: Intramuscular pressure, ultrafiltrate volume, ultrafiltrate and serum levels of creatine kinase and lactate dehydrogenase, histologic measurement of extracellular and intracellular edema, as well as the degree of cellular necrosis. RESULTS: Intramuscular pressure tended to be lower on the treated side at the end of the treatment period [treated leg: 9.3 +/- 4.0 mm Hg (+/- SE), control leg: 19.3 +/- 1.4 mm Hg, P = 0.03]. Analysis of ultrafiltrate fluid showed that levels of creatine kinase and lactate dehydrogenase were elevated compared with serum levels. Creatine kinase levels in serum were measured at 4150 +/- 780 U/L, whereas ultrafiltrate levels of creatine kinase were 28,700 +/- 17,700 U/L (+/- SE) (P = 0.1). Lactate dehydrogenase was measured at 1950 +/- 180 U/L in serum, but markedly elevated in ultrafiltrate [160,000 +/- 88,900 U/L (+/- SE), P = 0.05]. Quantification of cellular and interstitial dimensions showed no difference in control and experimental limbs. Quantification of the degree of muscle necrosis revealed 6.1 +/- 2.7% necrosis in the treated limb compared to 11.3 +/- 1.6% necrosis in the control group (P = 0.02, df = 2, 1-tailed paired t test). CONCLUSION: This pilot study demonstrates the feasibility of tissue ultrafiltration for reduction of intramuscular pressure in this porcine model. Further studies are underway. Compartment syndrome ultrafiltration may be useful prophylactically in patients at risk for acute compartment syndrome. Sampling of interstitial fluid and frequent measurement of intramuscular pressure may allow earlier diagnosis and treatment of acute compartment syndrome, whereas the reduction of tissue pressure by compartment syndrome ultrafiltration may prevent acute compartment syndrome from occurring. Additionally, compartment syndrome ultrafiltration will not hinder the ability of clinicians to use the clinical examination and pressure monitoring as the gold standard. PMID- 15795577 TI - Technical aspects of bridge plating for pediatric femur fractures. AB - Bridge plating for the adult orthopedic trauma population has been increasing over the last few years. The technique, used in comminuted fractures, allows for stable internal fixation while reducing blood loss and the need for fracture site exposure. We have applied the technique of bridge plating for comminuted diaphyseal femur fractures in the pediatric trauma population due to the limitations of fracture treatment options in these patients. The use of 4.5-mm narrow low-contact dynamic compression plate plates with minimal exposure for plate entry and percutaneous screw placement has greatly facilitated the treatment of these fractures. The technique creates a stable mode of fixation while maximizing biologic fracture healing potential and also permits early patient mobilization. PMID- 15795578 TI - Technique for removal of cannulated screws. AB - Cannulated screws can become incarcerated or stripped during the process of initial open-reduction internal fixation or at the time of hardware removal. In addition, many different sizes and brands of cannulated screws exist, and the appropriate size or type of screwdriver may not be available. We describe a simple technique for cannulated screw removal that works for all types of screws and can be performed percutaneously using only a Steinmann pin and T- handle chuck or pin driver. PMID- 15795579 TI - Traumatic dislocation of the hamate and pisiform: a case report and review of the literature. AB - Isolated dislocation of the hamate bone and pisiform bone is rare. We describe the simultaneous complete dislocation of both the hamate and pisiform bones in a 27-year-old man who crushed his right hand in a rolling press. An open reduction and internal fixation with Kirschner wires was performed. Four weeks later, the Kirschner wires were removed and rehabilitation was started. At 6 months follow up, the patient had minimal pain and full range of motion in the affected wrist joint and fingers. However, grip strength was 50% compared to his unaffected left hand, and sensation of the ulnar nerve area was reduced to almost 30% of that of his left hand. It appears that the ulnar nerve injury was the largest contributing factor to the poor outcome of our patient. Evaluation of soft-tissue injuries, especially nerve injury, is important in the treatment of complex carpal dislocations. PMID- 15795580 TI - Amputations through the hind foot: a report of three cases using a modified Dwyer and Paul procedure. AB - Salvage of limb length following oblique proximal foot amputations is presented. The technique, results, and follow-up of this procedure are described and compared with other standard procedures. A prospective study was done on 3 adolescents who presented consecutively during years 1999 and 2000. The salvage procedure described was performed on all 3 patients, and they were followed up for an average 3.5 years (range 3-4 years). All patients ambulated soon after surgery using crutches. Union of the graft between the distal tibia and calcaneus was achieved at 6 months, and all patients could walk bare foot as well as using their prosthesis both indoors and outdoors. We conclude that salvage of leg length and preservation of the sole of the foot in its normal plantigrade position by this procedure offers a viable and acceptable option to the surgeon and young patients. PMID- 15795581 TI - Spontaneous ejection of a ring sequestrum in a 10-year-old: a case report. AB - A case of a 10-year-old patient with a femur fracture treated with an external fixator who subsequently went on to develop a free-floating ring sequestrum is presented. The patient was largely asymptomatic throughout his course of treatment until seen emergently at 2 years postinjury with local and systemic signs of infection. He was successfully treated with intravenous and oral antibiotics as well as thorough irrigation and debridement. Ring sequestrum is the end point for pin tract infection that is left untreated. When it occurs, a course of antibiotics and thorough irrigation and debridement can lead to a successful outcome. PMID- 15795582 TI - The treatment of subtrochanteric nonunions with the long gamma nail: twenty-six patients with a minimum 2-year follow-up. PMID- 15795584 TI - Pediatric Gastroenterology Workforce Survey, 2003-2004. AB - BACKGROUND: The North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition (NASPGHAN) performed a workforce survey to determine the number, distribution, and work-related activities of pediatric gastroenterologists in the United States of America (USA) and Canada, and compared these findings with the first workforce survey completed in 1996. METHODS: The survey queried pediatric gastroenterologists in the USA and Canada between November, 2003 and June 2004. To permit the optimal comparison to the 1996 survey data, the original survey was used as a template for development of the current instrument and, when possible, the questions were left unchanged. Additional questions were added to address important contemporary issues not present in the initial survey. Limited income information was also collected. The survey was posted on the NASPGHAN website, and the NASPGHAN membership was notified of the survey by electronic mail via its electronic mail distribution list. This was followed by a three-part postal mail survey to all non respondents. After the Internet and postal mail requests, all non-respondent physicians were telephoned a minimum of three times. If unsuccessful in contacting the physicians directly, office personnel were queried to facilitate survey completion regarding the provision of pediatric gastroenterology, nutrition or hepatology services in either clinical care or research. RESULTS: The response rate based on the potential contact list for Part I of the survey was 69%. The final phone call or electronic mail contact of an office staff member with questions regarding gender and delivery of pediatric gastroenterology services yielded a total contact rate of 88%. There were 699 pediatric gastroenterologists identified in North America, as compared with 672 in 1996. If known non-respondents are included, there could be as many as 794 pediatric gastroenterologists. Time spent in clinical activities increased from 60% to 66% in the USA and from 43% to 53% in Canada. The use of nurse practitioners and physician assistants has increased considerably over the past 7 years. Fifty three percent of respondents feel there are too few pediatric gastroenterologists. Fifty percent of section and practice heads report that they are currently recruiting partners. Limited income information is presented. CONCLUSIONS: There is currently a self-perceived shortage of pediatric gastroenterologists as compared with 7 years ago, despite a constant proportion of pediatric gastroenterologists per million children. In the USA, nurse practitioners and physician assistants are being increasingly used to fill this need, and physicians in both Canada and the USA have increased the time they spend in clinical care. PMID- 15795585 TI - Digestive endoscopy in neonates. AB - Since the introduction of flexible fiberoptic endoscopy in the early 1970s, esophagogastro-duodenoscopy and colonoscopy have become established procedures for the diagnosis, evaluation and treatment of gastrointestinal tract disease in the pediatric population. The development of safe fiberoptic endoscopes specially designed for neonates has allowed visualization of lesions occurring in the first days of life. Despite an increased understanding of neonatal digestive disorders deriving from this new diagnostic modality, there is little consensus on the appropriate use of endoscopic procedures in routine care of neonates. It is the feeling of the authors that widening the indications of endoscopy in the neonatal period might lead to diagnosis of discrete clinical abnormalities, which might improve the care of neonates. The techniques for performing neonatal endoscopies, the appropriate indications, the common normal and pathologic findings and the complications of these procedures are reviewed. PMID- 15795586 TI - Celiac disease: to screen or not to screen the general population? PMID- 15795587 TI - Orthostatic intolerance and the biopsychosocial model. PMID- 15795588 TI - Gastrointestinal symptoms associated with orthostatic intolerance. AB - BACKGROUND: The term orthostatic intolerance is used to describe symptoms of hemodynamic instability such as lightheadedness, fatigue, impaired cognition and syncope that develop on assuming an upright posture. Common forms of orthostatic intolerance in childhood include postural tachycardia syndrome and neurally mediated hypotension. OBJECTIVE: A descriptive report of the clinical characteristics of patients presenting with gastrointestinal symptoms who are ultimately found to have orthostatic intolerance. METHODS: A medical record review of all patients referred to the pediatric gastroenterology service at the Johns Hopkins Children's Center who had an abnormal tilt table test between June 1996 and December 2000. RESULTS: Of 24 eligible subjects aged 9-17 years (mean, 14.3 years), four had postural tachycardia syndrome, eight had both postural tachycardia and neurally mediated hypotension, and 12 had neurally mediated hypotension alone. The most common presenting gastrointestinal symptoms were abdominal pain, nausea and vomiting. Median number of gastrointestinal symptoms per patient was 3 (range, 1-7), and 87% of the patients experienced gastrointestinal symptoms for more than 1 year and 48% experienced gastrointestinal symptoms for more than 3 years. Follow-up information was available on 18 patients. Seventy-eight percent of patients (14 of 18) had complete resolution of symptoms with treatment of orthostatic intolerance. CONCLUSION: Pediatric patients with chronic upper gastrointestinal symptoms may have underlying orthostatic intolerance. In patients with upper gastrointestinal symptoms and orthostatic intolerance, treatment of orthostatic intolerance may result in resolution of gastrointestinal symptoms. PMID- 15795589 TI - Gastroesophageal reflux in familial dysautonomia: correlation with crisis frequency and sensory dysfunction. AB - OBJECTIVES: Familial dysautonomia (FD), a genetic disorder resulting in sensory and autonomic nervous system dysfunction, is associated with a high frequency of gastroesophageal reflux (GER). In this study the authors attempted to determine if GER prevalence correlated with dysautonomic crisis frequency and small fiber sensory dysfunction. METHODS: Of an initial population of 252 FD patients who had survived past their seventh year and were alive during the year of study from June 1, 2001 to June 1, 2002, 174 (69.0%) had sufficient data in their patient files to be included in this retrospective analysis. Subjects were stratified into three groups: those with no GER, those medically treated for GER and those surgically treated for GER with fundoplication. Data were obtained regarding frequency and severity of dysautonomic crises and sensory function including pain sensation, cold and warm thresholds and vibration perception. RESULTS: There were no significant differences among the three groups regarding age or severity of sensory dysfunction. However, dysautonomic crises were significantly more frequent in FD patients who were presumed to have more intractable GER, as they had required fundoplication (P < 0.05). CONCLUSIONS: The data confirm the high incidence of GER in the FD population and demonstrate that GER is independent of severity of sensory dysfunction. In addition, the authors demonstrate that fundoplication does not alleviate the central autonomic cause of retching, the dysautonomic crisis. PMID- 15795590 TI - Assessment of the rectoanal inhibitory reflex in preterm infants with delayed meconium passage. AB - BACKGROUND: There is an inverse relationship between gestational age, birth weight and the time of first neonatal bowel movement. The authors hypothesized that delayed passage of meconium might result from a delayed maturation of the recto-anal inhibitory reflex (RAIR) in premature infants. OBJECTIVE: To evaluate whether the RAIR is absent in very preterm infants 28-32 weeks postmenstrual age with delayed meconium production. STUDY DESIGN: Anorectal manometry was performed in 10 preterm infants (seven male) with delayed meconium production (no meconium in the first 48 hours). Median postmenstrual age was 30 weeks (28-31 weeks). Birth weight ranged from 780 to 1930 g (median, 1395 g). A micromanometric assembly (outer diameter, 2.0 mm) was used which incorporated a 1.5-cm-long sleeve sensor for measurement of resting anal sphincter pressure and relaxation. Four side-holes recorded anal and rectal pressures. Rectal distension was performed with direct air insufflation to elicit the RAIR. RESULTS: The time from birth to passage of meconium ranged from 48 to 105 hours (median, 82 hours). The mean anal sphincter pressure, rectal pressure, and anal sphincter oscillation frequency were 22.0 +/- 5.0 mm Hg, 6.9 +/- 2.0 mm Hg, and 9.8 +/- 1.9/min, respectively. A normal RAIR was elicited in all infants. CONCLUSION: Anorectal manometry recordings in premature infants with delayed passage of meconium showed normal anorectal pressures and a normal RAIR, suggesting that delayed meconium passage is not related to the absence of a RAIR. PMID- 15795591 TI - Fecal elastase: pancreatic status verification and influence on nutritional status in children with cystic fibrosis. AB - OBJECTIVES: To evaluate fecal elastase 1 (FE) levels in young children with cystic fibrosis and pancreatic insufficiency and to explore the relationship between FE and growth, nutrition, pulmonary status and fat absorption over a 24 month period. METHODS: FE, indicating pancreatic lipase activity, was assessed in children (6.0 to 8.9 years of age) with cystic fibrosis and pancreatic insufficiency. FE >15 microg/g stool was defined as residual pancreatic activity, suggesting some pancreatic function, and FE 0.990 and slope displaying an 8.8 interassay %CV. At the lower limit of quantification, 2 ng/mL, the percentage bias and %CV were -5.0% and 14.7%, respectively. Intraassay precision at weighed-in levels of 6, 12, and 32 ng/mL were 2.4% to 6.4%, and biases were -10.7% to -8.5%. These same quality control materials yielded -6.3% to -0.8% biases from the expected values and 2.4% to 10.9% interday precision, respectively. This method for everolimus determination, validated according to FDA guidelines, provides longer column life and better sensitivity than that of French et al for sirolimus determination. This protocol also provides acceptable accuracy and precision over the expected therapeutic range and allows 1 technologist using 1 LC/UV system to run up to 5000 samples per year with confidence. PMID- 15795649 TI - Do thyroxine and thyroid-stimulating hormone levels reflect urinary iodine concentrations? AB - The toxicity of environmental chemicals such as nitrates, thiocynates, and perchlorates, some therapeutics, and dietary goitrogens can lower thyroidal iodine uptake and result in hypothyroidism and goiter. Iodine sufficiency, essential for normal thyroid hormone synthesis, is critical during gestation to assure that sufficient thyroxine (T4) and iodine reach the developing fetus. Spot urinary iodide (UI) measurements are used globally to indicate and monitor iodine sufficiency of populations. In individuals, however, UI are not routinely measured; instead, normal serum thyroid-stimulating hormone (TSH) and T4 concentrations serve as surrogate indicators of iodine sufficiency as well as thyroidal health. Our objective was to examine the relationship between UI concentrations and serum T4 and TSH concentrations in individuals in an "iodine sufficient population." Using a cross-sectional sample of the US population (n = 7628) from the National Health and Nutrition Examination Survey (NHANES III; 1988 1994) database, we examined the relationship among UI, T4, and TSH in pregnant and nonpregnant women and in men (15-44 years). There was a lack of relationship between UI (or UI/Cr) concentrations and serum T4 or TSH concentrations. Therefore, TSH and T4 are not appropriate markers of UI concentrations in this population. Monitoring the status of iodine nutrition of individuals in the United States may be important because serum TSH and T4 concentrations do not indicate low iodine status. PMID- 15795650 TI - Simple and simultaneous determination of the hiv-protease inhibitors amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir plus M8 nelfinavir metabolite and the nonnucleoside reverse transcriptase inhibitors efavirenz and nevirapine in human plasma by reversed-phase liquid chromatography. AB - Several studies suggest that therapeutic drug monitoring of protease inhibitors and nonnucleoside reverse transcriptase inhibitors may contribute to the clinical outcome of HIV-infected patients. Because of the growing number of antiretroviral drugs and of drug combinations than can be administered to these patients, an accurate high-performance liquid chromatographic (HPLC) method allowing the simultaneous determination of these drugs may be useful. To date, the authors present the first simultaneous HPLC determination of the new protease inhibitor atazanavir with all the others currently in use (M8 nelfinavir metabolite included) and the 2 widely used nonnucleoside reverse transcriptase inhibitors efavirenz and nevirapine. This simple HPLC method allows the analysis all these drugs at a single ultraviolet wavelength following a 1-step liquid-liquid extraction procedure. A 500-muL plasma sample was spiked with internal standard and subjected to liquid-liquid extraction using by diethyl ether at pH 10. HPLC was performed using a Symmetry Shield RP18 and gradient elution. All the drugs of interest and internal standard were detected with ultraviolet detection at 210 nm. Calibration curves were linear in the range 50-10,000 ng/mL. The observed concentrations of the quality controls at plasma concentrations ranging from 50 to 5000 ng/mL for these drugs showed that the overall accuracy varied from 92% to 104% and 92% to 106% for intraday and day-to-day analysis, respectively. No metabolites of the assayed compounds or other drugs commonly coadministered to HIV-positive patients were found to coelute with the drugs of interest or with the internal standard. This assay was developed for the purpose of therapeutic monitoring (TDM) in HIV-infected patients. PMID- 15795652 TI - 10-Hydroxycarbazepine serum concentration-to-oxcarbazepine dose ratio: influence of age and concomitant antiepileptic drugs. AB - This study was done to evaluate the association between patient age and the concomitant use of enzyme-inducing antiepileptic drugs (AEDs) and oxcarbazepine (OXC) concentration-to-dose ratio (CDR) by a multivariate analysis. The influence of patient age and concomitant AEDs on the trough steady-state serum concentration of 10-hydroxycarbazepine (OHC) normalized to 1 mg/kg body weight of OXC or concentration-to-dose ratio (OHC-OXC-CDR) was assessed by analysis of covariance. Samples were collected from 106 patients (90% outpatients), aged 1 80, who were receiving OXC either alone (n = 41) or in combination with other AEDs (n = 65). The average OHC-OXC CDR was 0.70 +/- 0.26 (mean +/- SD). Analysis of covariance showed that patient age was influential (P < 0.001) and that there was a difference between the noninducers group (OXC or OXC + lamotrigine, topiramate, or valproate) and the inducers group (OXC + phenobarbital or phenytoin) (P < 0.001). The OHC-OXC CDR increased with age (r = 0.14, P < 0.001) and was approximately 48% lower in children aged 6 or less than in patients over 45, and approximately 32% lower in the inducers group than in patients receiving OXC alone. The correlation between OHC-OXC CDR and the age of the patients concerned with OXC alone was r = 0.48, P < 0.001. In the noninducers group the OHC-OXC CDR was 0.59 +/- 0.24 in patients aged 11 or less (n = 16), and 0.81 +/- 0.23 in patients over 11 years (n = 62). In the inducers group it was 0.25 +/- 0.11 in patients aged 11 or less (n = 3) and 0.57 +/- 0.18 in patients over the age of 11 (n = 25). The OHC-OXC CDR increased with patient age and decreased in the presence of enzyme-inducing AEDs in epileptic patients chronically treated with OXC. These influences may be clinically relevant, and, therefore, patient age and the presence of inducers should be considered in estimating either compliance or the OXC dose needed to achieve a desired OHC concentration. PMID- 15795651 TI - Lack of clinically significant pharmacokinetic interactions between zonisamide and lamotrigine at steady state in patients with epilepsy. AB - The present study was designed to assess the effect of the addition of zonisamide (ZNS) on lamotrigine (LTG) disposition and the safety of the combination under steady-state conditions in patients with epilepsy. A secondary objective was to characterize ZNS pharmacokinetics (PK) in the presence of LTG. Twenty subjects (male and female 18 to 55 years old) stabilized on LTG monotherapy (150-500 mg/d) took part in a 2-center, open-label, 1-way drug interaction study. ZNS was gradually increased to 200 mg twice daily over a 3-week period, and 3 PK profiles were performed: on days -7 and -1 to assess the PK of oral LTG administered alone and on day 35, after 14 days of ZNS at the maximal tolerated dose, to evaluate the effect of ZNS on LTG PK and to characterize ZNS PK in the presence of LTG. Eighteen subjects completed the study. Steady-state dosing of ZNS did not significantly affect the mean Cmin (mean +/- SD: 2.8 +/- 1.4 vs 3.5 +/- 2.4 microg/mL), Cmax (5.1 +/- 3.0 vs 5.1 +/- 3.0 microg/mL), AUC0-12 (45.5 +/- 22.6 vs 50.3 +/- 32.1 microg.h/mL), and CL/F (2778.5 +/- 1368.5 vs 3052.1 +/- 2744.9 mL/h) of LTG measured before (day -1) and after (day 35) ZNS administration, respectively. Further, 90% confidence intervals for the geometric mean ratios (day 35/day -1) fell within the no-effect range of 0.80-1.25. The fraction of dose of total and unconjugated LTG excreted in urine was not significantly different between baseline and ZNS treatment, but the renal clearance of LTG decreased significantly with ZNS dosing (P = 0.01). On the other hand, the PK parameters measured for ZNS in the presence of LTG were consistent with an absence of LTG effect on ZNS PK. The coadministration of ZNS and LTG was generally well tolerated. Steady-state safety and PK of LTG and ZNS are not affected significantly when these two drugs are coadministered at clinically relevant doses, indicating that no dosage adjustment of either drug should be required when they are used in combination. PMID- 15795653 TI - Misleading high tobramycin plasma concentrations can be caused by skin contamination of fingerprick blood following inhalation of nebulized tobramycin (TOBI): a short report. AB - We observed unexpected high plasma concentrations of tobramycin (48.5 and 28.1 mg/L) in fingerprick blood samples after the nebulization of tobramycin solution for inhalation (tobramycin 300 mg/5 mL, TOBI by 2 young children aged 3 years. To investigate whether dermal contamination could be the source of error, 3 adult volunteers were present during another nebulization by a third child (age 2 years). The volunteers had exposure to tobramycin by handling the nebulizer or the nebule and also by inhalation from holding the child and being in close proximity while TOBI was being administered. Five blood samples by fingerprick and 2 by venipuncture were collected and assayed for tobramycin concentration. On each occasion the site was swabbed with alcohol wipes to mimic standard patient sampling methods. One site was resampled after cleaning of hands with 2% chlorhexidine gluconate and water. Tobramycin concentrations from venipuncture 1 2 hours after nebulization were all <0.2 mg/L except for 1 result of 1.2 mg/L. The tobramycin concentrations from fingerpricks before hand washing varied between 6.8 and 172 mg/L, and after hand washing between 0.3 and 17.6 mg/L. Contamination of fingers with tobramycin is likely to have caused the error in the 2 initial cases and did cause misleadingly elevated levels in the adult volunteers. We caution that therapeutic drug monitoring of nebulized tobramycin should not be done by fingerprick sampling, and care should be taken to avoid contamination of the venipuncture site. PMID- 15795654 TI - Genetic polymorphism of CYP2C9 in a Vietnamese Kinh population. AB - Cytochrome P450 2C9 (CYP2C9) shows genetic polymorphism with high interethnic variation, but no report has addressed the genetic polymorphism in the Vietnamese population. In the present study, the distribution of 2 common allelic variations of CYP2C9 was investigated in Vietnamese Kinh population, a major ethnic group in Vietnam. Genomic DNA from 157 Vietnamese subjects was amplified by polymerase chain reaction, and the presence of CYP2C9*2 and CYP2C9*3 allelic variants was determined by pyrosequencing. Among 157 Vietnamese subjects, no subject with the CYP2C9*2 allele was detected, but 7 subjects were heterozygous for the CYP2C9*3 allele. The allele frequency of CYP2C9*3 was 2.2% in the Vietnamese Kinh population. This genotype distribution was well correlated with previous reports suggesting no occurrence of CYP2C9*2 in Asians. These results suggest that CYP2C9*2 may be absent in Vietnamese Kinh population and that CYP2C9*3 is major allelic variant that causes interindividual variation of drug responses to CYP2C9 substrate drugs in the Vietnamese Kinh population. PMID- 15795655 TI - [Anti-hepatitis B vaccination: the hepatologist's point of view]. PMID- 15795656 TI - Carbohydrate-deficient transferrin and gamma-glutamyl transpeptidase in the evaluation of alcohol consumption. A five-year retrospective study of 633 outpatients in a single center. AB - OBJECTIVE: Carbohydrate-deficient transferrin has been proposed to be useful in evaluating alcohol consumption but there is no consensus on its use in routine practice. The aim of this retrospective study was to compare carbohydrate deficient transferrin and gammaglutamyl transpeptidase assays for the evaluation of alcohol consumption. METHODS: Six hundred thirty-three outpatients attending one outpatient care center were included in this study. Patients were divided into five categories according to alcohol consumption: category 1 included non weaned patients drinking more than 30 g/day for women and more than 50 g/day for men, category 2 included relapse patients, category 3 included moderate drinkers, category 4 included patients weaned less than one month, and category 5 included patients weaned more than one month. One experienced physician estimated alcohol intake from patient declarations during a face-to-face interview. RESULTS: Sensitivity of carbohydrate-deficient transferrin varied, depending on patient category, from 32% to 92% versus 41% to 72% for gamma-glutamyl transpeptidase. Specificity of carbohydrate-deficient transferrin varied from 71% to 96% versus 23% to 62% for gamma-glutamyl transpeptidase. After one month of abstinence, specificity of carbohydrate-deficient transferrin was 62% versus 19% for gamma glutamyl transpeptidase. CONCLUSION: This study confirms that carbohydrate deficient transferrin is more accurate in predicting alcohol consumption compared with gamma-glutamyl transpeptidase in alcoholic outpatients. PMID- 15795657 TI - Treatment of chronic hepatitis C in patients unresponsive to interferon. Interest of re-treatment combining interferon induction therapy and ribavirin (a multicenter pilot study). AB - AIM: About 45% of patients with chronic hepatitis C are unresponsive to the present reference treatment combining pegelated interferon plus ribavirin; before pegylated interferon was available the non-response rate was around 60%. This open multicenter pilot study, initiated before pegylated interferon became available, was designed to evaluate, in patients unresponsive to interferon monotherapy, the rate of biological and virological response and side-effects of the ribivirin-alpha 2b interferon combination. METHODS: The combination protocol was ribavirin (1 to 1.2 g/d) plus alpha 2b interferon at induction doses (9 MU/d the first week; 4.5 MU/d the eleven following weeks; 3 MU/2 days the 36 following weeks). RESULTS: Among the 27 included patients, 17 (63%) were viremia-negative (PCR) after 12 weeks of treatment, 9 (33%) were complete responders (undetectable viremia and normal transaminases) at the end of treatment (48 weeks) and of follow-up (72 weeks). Patients with non-1, non-4 genotypes who derived full benefit from this therapeutic strategy (6/7 (86%) were complete responders: 4/5 with genotype 3 and 2/2 with genotype 5). Quality-of-life was impaired during treatment, especially during the first 12 weeks of high-dose interferon therapy. CONCLUSION: While waiting for new therapeutic possibilities, these good results suggest interferon induction at the beginning of treatment remains a valid option. PMID- 15795659 TI - [Coloscopic hepatitis C virus contamination]. PMID- 15795660 TI - [Proton pump inhibitors and upper gastrointestinal bleeding]. PMID- 15795658 TI - [Psychiatric disorders during treatment of chronic hepatitis C]. PMID- 15795661 TI - Management of bleeding peptic ulcer in France: a national inquiry. AB - AIMS OF THE STUDY: To evaluate and compare management practices in France for bleeding peptic ulcers using a national inquiry of university and non-university hospitals. METHOD: Responses to questionnaires sent to 812 gastroenterologists, 496 practicing in non-university hospitals and 316 in university hospitals, were compared. RESULTS: An analysis was possible in 279 (34% response rate) of the questionnaires. Forrest classification was used more frequently in university hospitals (83% vs 60%, P<0.01). Endoscopic hemostatic therapy was used more frequently in university hospitals for Forrest Ib (92% vs 81%, P=0.02), IIa (93% vs 73%, P<0.001), and IIb (58% vs 29%, P<0.001) ulcers. Injection therapy, mainly epinephrine, was the first-intention treatment for 99% of the responding gastroenterologists. Proportions of clinicians employing hemoclips (27%) or argon plasma coagulation (21%) were similar in both types of practice. Anti-secretory treatment included mainly omeprazole (82%), given intravenously (76%), sometimes as bolus i.v. doses followed by i.v. high-dose continuous infusion (15%) with some variations according to the type of hospital. In the event of recurrent or persistent bleeding, surgery was more frequent in non-university hospitals. When rebleeding occurred, a second endoscopic treatment was performed in about one quarter of patients. CONCLUSION: In France, management practices for bleeding peptic ulcer vary between university and non-university hospitals. PMID- 15795662 TI - Predictors of response to infliximab in luminal Crohn's disease. AB - AIMS: To identify predictive factors of response to infliximab in luminal Crohn's disease (CD). PATIENTS AND METHODS: All consecutive patients with luminal CD treated with infliximab between October 1999 and March 2003 in Bordeaux's referral centers were included. All had at least 3 months follow-up post infliximab infusion and no prior treatment with infliximab. Response rates were determined 2 and 8 weeks after infusion according to Crohn's Disease Activity Index (CDAI) (remission=CDAI<150 and response=CDAI decrease more than 100). RESULTS: Among 44 patients (33 female; mean age 35 +/- 14 yr.), 39 (88%) had a clinical response 2 weeks after infusion (79% in remission). At week 8, the rate of response was 61.4% and exclusive colonic involvement predicted sustained response to treatment (P=0.03). The probability of remission at 56 weeks was 21.4%. Multivariate analysis demonstrated that the only factor associated with response duration was initiating immunosuppressive (IS) therapy in women (RR=3.61 95%CI[1.25-10.41], P=0.017). CONCLUSION: Exclusive colonic involvement is the only predictive factor of sustained response to infliximab in luminal CD. At the time of infliximab infusion, initiation or modification of IS therapy may favor sustained response, at least in women. PMID- 15795663 TI - Accuracy of preoperative imaging methods in a retrospective series of 14 patients with operated intraductal papillary mucinous tumors of the pancreas. AB - Accuracy of preoperative imaging methods for the diagnosis of intraductal papillary mucinous tumor of the pancreas (IPMT) is not well known. AIMS: To compare diagnostic accuracy of various preoperative imaging methods with pathology data following surgical resection. PATIENTS AND METHODS: Fourteen consecutive patients underwent pancreatic surgical resection for IPMT between January 1988 and May 2002. Imaging methods included endoscopic retrograde cholangiopancreatography and/or magnetic resonance cholangiopancreatography and/or endoscopic ultrasonography. Results of preoperative morphological examinations were compared with histopathological findings from surgical specimens. RESULTS: IPMT was located to the pancreatic head or body in 72% of patients and involved the main pancreatic duct (MPD) in 79% of cases Carcinoma was diagnosed in 35% of the cases. Sensitivity of imaging methods varied from 64 to 80% to accurately locate the tumor and from 73 to 80% in distinguishing between types involving the MPD or the accessory ducts. Planned surgical resection based on preoperative imagery correlated with final surgery in 57% of the patients. Histological study of whole pancreas specimens revealed lesions in undilated ducts, and also dilated ducts without histological lesions. CONCLUSION: The sensitivity of preoperative imaging methods is moderate in defining intraductal extension of IPMT. Duct dilatation is not predictive of histological involvement by tumors and ducts may be pathological without dilatation. PMID- 15795664 TI - [Migraines and digestive disorders]. PMID- 15795666 TI - [Management of anoperineal lesions in Crohn's disease]. PMID- 15795667 TI - [Anoperineal lesions in Crohn's disease: an ongoing story]. PMID- 15795669 TI - [Interview: Questions to Jean-Francois Contou]. PMID- 15795670 TI - Percutaneous treatment of a complex hydatid cyst of the liver under sonographic control. Report of the first case. AB - Surgical procedures are considered the gold standard treatment of hydatid cysts. We report the first case of percutaneous treatment of a complex hydatid cyst of the liver, using radio frequency as a scolecidal agent in a 28-year-old man. The procedure was performed under real-time ultrasound control and with the patient under general anesthesia. Injection of alcohol was performed at the end of the procedure to induce retraction of the residual cavity. The percutaneous procedure was associated with oral albendazole therapy. Six months after the procedure, the patient was in very good condition. Liver and blood tests were normal and the cyst cavity was reduced. Further studies are required to define the indication of this new therapeutic strategy of hydatid cyst. PMID- 15795672 TI - [Osteoclastic giant cell tumour of the pancreas]. AB - Osteoclast giant cell tumours are bone tumours that occur in adults, and that are considered benign by WHO but locally aggressive. Strictly identical tumours are described in the pancreas, without simultaneous bone localization. We report the case of a 62-year woman with an osteoclast giant cell tumour of the distal pancreas, without any epithelial component, which was diagnosed after pancreatic resection and with no signs of recurrence after a 24-month follow-up. These pancreatic tumours are rare, with a very poor prognosis, an unclear histogenesis; they are often confused with pleomorphic or undifferentiated pancreatic carcinomas including a component of osteoclast giant cell. These osteoclast giant cell tumours of the pancreas usually present as large cystic tumours. In certain cases, complete resection can result in long-term survival. PMID- 15795671 TI - [Crohn's disease associated with interferon and ribavirin treatment for chronic hepatitis C]. AB - Digestive side effects associated with interferon and ribavirin in the treatment of chronic hepatitis C seem to be non specific. So far, inflammatory bowel diseases have only been rarely described in this context. We report two cases of Crohn's disease which occurred a few months after interferon and ribavirin treatment for chronic hepatitis C. The first case was a 43 year old man treated for 7 months with a interferon and ribavirin combination for chronic hepatitis C who experienced colitis related to Crohn's disease, with the occurrence of articular and cutaneous manifestations. The second case was a 43 year old women treated for 8 months with a pegylated interferon and ribavirin combination for chronic hepatitis C who developed Crohn's disease with right ileo-colic topography and severe evolution requiring an ileo-coecal resection because of the failure of medical treatment. The alpha interferon used for its antiviral action against hepatitis C virus, has complex immuno-modulation effects which could reveal latent Crohn's disease. PMID- 15795674 TI - [Gangrene of the ligamentum teres hepatis]. AB - Necrosis of the ligamentum teres hepatis is a very rare and misdiagnosed pathology. Four cases have been reported in the literature. Clinical symptoms with acute abdominal pain suggest peritonitis. The diagnostic may be obtained by CT scan of the abdomen revealing a hypo/dense circular lesion, anterior to the liver, to the junction of the segments III and IV, associated with inflammation. The mechanism of isolated necrosis of the round liver ligament remains unclear. In the absence of a preoperative diagnosis, a video-laparoscopic exploration is proposed. PMID- 15795673 TI - [Cancer of an ectopic pancreas in the duodenal wall]. AB - Ectopic pancreas is defined as pancreatic tissue outside the normal location of the pancreas. It can be affected by the same complications as the orthotopic pancreas, such as adenocarcinoma. This extremely rare complication (only 14 published cases) may have a better prognosis that adenocarcinoma of an orthotopic pancreas. Endoscopic ultrasonography may be useful in the diagnosis of this disease. We report a case of malignancy of the duodenal wall originating in aberrant pancreatic tissue, and review the literature. PMID- 15795675 TI - [Cirrhosis in children and SZ phenotype alpha1-antitrypsin deficiency]. PMID- 15795676 TI - [Role of TTF-1 immunohistochemistry for the diagnosis of duodenal metastasis of pulmonary adenocarcinoma]. PMID- 15795677 TI - [Small bowel adenocarcinoma revealing Crohn's disease]. PMID- 15795678 TI - [Granular cell tumor involving the appendix]. PMID- 15795679 TI - [About "Is routine pathologic evaluation of hemorrhoidectomy specimens necessary?"]. PMID- 15795681 TI - [Wirsung: a name before becoming an anatomical reference]. PMID- 15795682 TI - Elidel and protopic. PMID- 15795683 TI - Erlotinib (Tarceva) for advanced non-small cell lung cancer. AB - Erlotinib (Tarceva) is the second oral epidermal growth factor receptor (EGFR) inhibitor to become available in the US for treatment of advanced refractory NSCLC. In clinical trials, erlotinib produced a response rate of only 8.9%, but increased median survival from 4.7 to 6.7 months. Patients who had never smoked and those with EGFR-positive tumors survived longer. Erlotinib is generally well tolerated; diarrhea and rash are the most common adverse effects. PMID- 15795684 TI - Extended-release carbamazepine (Equetro) for bipolar disorder. AB - An extended-release formulation of carbamazepine, available since 1997 for treatment of epilepsy, has now been approved under a new name, Equetro, for acute mania and mixed episodes of bipolar disorder. Although the drug was effective in some patients for up to 6 months, it has not been approved for maintenance treatment. Carbamazepine has not been shown to be more effective than lithium or valproate, and it can cause serious adverse effects. PMID- 15795685 TI - Adderall. PMID- 15795686 TI - Toxicity induced by Hg2+ on choline acetyltransferase activity from E. electricus (L.) electrocytes: the protective effect of 2,3 dimercapto-propanol (BAL). AB - BACKGROUND: The effect of mercury (Hg(2+)) on the activity of choline acetyltransferase (ChAT) from electrocytes of Electrophorus electricus (L.) was studied due to the importance of this enzyme and acetylcholine in many neurochemical functions such as arousal, learning, and memory. MATERIAL/METHODS: Mercury, which has affinity to thiol groups, acted as a potent inhibitor of ChAT, which was obtained by differential centrifugation and ammonium sulfate precipitation, at 80%, from the main electric organ homogenate. RESULTS: Mercury inhibition presents different kinetic behaviors for both enzyme substrates: noncompetitive to choline and of mixed type to AcCoA, with inhibition constants on the order of 0.5 to 1.0 microM. The enzyme activity was recovered using 2,3 dimercapto-propanol (BAL), a well-known chelate for sulphydryl groups and metals, which acted as a protecting agent and was able to revert the Hg(2+) inhibition at a concentration of 10 (-6) M. After treatment with this metal and in the presence of 2,3 dimercapto-propanol, 70% of the enzyme activity was recovered for AcCoA and 80% for choline. CONCLUSIONS: The observed inhibition is likely due to direct protein interaction, because the addition of BAL reversed the effects of HgCl(2) on ChAT activity. The results cast new light on the mechanisms of mercurial neurotoxicity. PMID- 15795687 TI - Intracerebroventricular administration of thymoquinone, the major constituent of Nigella sativa seeds, suppresses epileptic seizures in rats. AB - BACKGROUND: Recently we investigated some neuropharmacological aspects of thymoquinone, such as anticonvulsant, muscle relaxant, and hypnotic effects, as well as its effect on motor coordination and locomotor activity. In this study, we evaluated the effect and mechanism(s) of the action of thymoquinone more precisely via intracerebroventricular (i.c.v.) injection. MATERIAL/METHODS: The anticonvulsant effects of thymoquinone, the major constituent of Nigella sativa seeds, were investigated using the pentylenetetrazole (PTZ)-induced seizure model. The animals were placed individually in plastic boxes and observed immediately after PTZ injection for a period of 30 min. The latency to and the duration of tonic-clonic seizures were recorded, as well as the percentages of protection against the incidence of seizure and mortality. RESULTS: In PTZ induced epileptic seizures, the i.c.v. injection of thymoquinone at doses of 200 and 400 microM prolonged the time until onset and reduced the duration of tonic clonic seizures. The protective effect of thymoquinone against lethality was 45% and 50% in the respective doses. In this study, flumazenil (1 nM, i.c.v.) reversed the anticonvulsant activity of thymoquinone. Also, pretreatment with naloxone (10 microM, i.c.v.) antagonized the prolongation of tonic-clonic seizure latency as well as the reduction in seizure duration induced by thymoquinone (200 microM, i.c.v.). CONCLUSIONS: These results indicate that thymoquinone may have anticonvulsant activity, probably through an opioid receptor-mediated increase in GABAergic tone. PMID- 15795688 TI - Effects of dental implant surfaces on the expression of bone sialoprotein in cells derived from human mandibular bone. AB - BACKGROUND: Protein adsorption is believed to be the first event that takes place after contact of natural tissue with an artificial surface. Bone sialoprotein (BSP) is one of the major noncollagenous proteins in the extracellular matrix of bone. The expression of BSP coincides with initial bone mineralization and is believed to be a center of crystallization for hydroxyapatite formation. MATERIAL/METHODS: We used a variety of four differently designed dental implant surfaces (SLA, CPT, ANOX, TICER) to investigate the effects on the development of adult human mandibular bone at days 5, 10, 15, 20, and 25 in vitro. The time course of the expression of BSP and labeling of fibroblasts was visualized immunohistochemically. The distribution patterns of cells were determined semiquantitatively on both the surface and the tissue-implant borderline. RESULTS: BSP immunoresponse increased from day 5 before decreasing after day 15 in vitro. The distribution of BSP-expressing cells changed during that time. Cell counts revealed that the time course of the settlement of cells depended on the design of the surface of the implant. The design of the border of the implant affected both the cell distribution patterns and the survivals of cells to a higher degree than did the design of the implant surface. CONCLUSIONS: Investigation of novel biomaterials for bone engineering represents an essential area for the design of tissue-engineering strategies. The hydroxyapatite-based implant material TICER could be a good scaffold to guide and promote the regeneration of bone tissue. PMID- 15795690 TI - Diagnosis of Lyme borreliosis using enzyme immunoanalysis. AB - BACKGROUND: Antiborrelia antibodies in Lyme borreliosis (LB) are mostly detected by enzyme immunoassay (EIA), confirmed by immunoblot (the "two-step system"). In indicated cases, direct evidence of Borrelia burgdorferi is obtained with the PCR method, electron microscopy and cultivation. The "one-step system" of testing for IgM and IgG antibodies in LB is economically preferably, but it requires an EIA kit with more than 90% sensitivity and specificity. MATERIAL/METHODS: 90 blood samples were collected, 54 from patients with clinically defined LB and 36 samples from individuals free of LB. IgM and IgG antibodies against Borrelia burgdorferi were detected in parallel with five different EIA kits from various producers. The results were verified clinically in all cases, in disputable cases with additional immunoblot (BAG-Med), and analyzed statistically. RESULTS: Specificity and sensitivity were calculated from the measured values, and diagnostic efficiency was determined for each EIA kit. EIA kits for antiborrelia antibody assay with high specificity have low sensitivity and vice versa. In 9 samples from patients with clinical diagnoses (multiple sclerosis, Parkinson disease, epilepsy, rheumatoid arthritis) we found false positives in EIA and WB tests. CONCLUSIONS: The best results for a "one-step system" of examinations for antiborrelia antibodies were obtained with the Abbot and Euroimmun EIA kits in our set. A "two-step system" of serological examination could be composed from the basic IgM and IgG examination with a high sensitivity EIA kit (Viroimmun, Test-Line) followed with confirmation of positives by specific immunoblot. PMID- 15795689 TI - Osteoinductive properties of urothelium depend on inherent bone morphogenetic proteins (BMPs). AB - BACKGROUND: The mechanism of heterotopic ossicle formation induced by grafted transitional epithelium, described in several species, has not yet been elucidated. This phenomenon is not species specific, but in xenogeneic systems, immunosuppression of the host is needed to protect the implant long enough for it to exert its osteoinductive potency. We showed a strong correlation between the intensity of bone induction caused by implantation of HeLa cell lines and their ability to synthesize and secrete seven different isoforms of BMPs (bone morphogenetic proteins). We decided to discover whether the urothelium of guinea pig, having high osteoinductive potency, differs in BMP content from human urothelium, which does not induce heterotopic bone. MATERIAL/METHODS: The RT-PCR technique was used to investigate the expression of BMP family genes. PCR products were directly sequenced with an ABI Prism 310 sequencer. The obtained sequences were analyzed using BLAST. RESULTS: Two isoforms of BMP (BMP-3 and BMP 4) were detected in the guinea pig urinary bladder mucosa. The sequences of these products were identified by BLAST analysis and compared with human mRNA and protein. The experiment performed on human urinary bladder mucosa failed to detect the expression of these BMPs, while b-actin mRNA was detected. CONCLUSIONS: Our findings explain the reason for the osteoinductive properties of guinea pig urothelium and are in line with the explanation of the osteoinductive properties of HeLa cells. Moreover, human urothelium, which did not express any BMP mRNA, therefore probably lacks osteoinductive competence. PMID- 15795691 TI - Treatment and prophylaxis with sucralfate ameliorates hypoxia/reoxygenation induced intestinal injury in pup rats. AB - BACKGROUND: Sucralfate is widely used as a cytoprotective agent in patients with peptic ulcer and other intestinal mucosal injury. The aim of this study is to investigate whether sucralfate has any effect on the prevention and treatment of hypoxia/reoxygenation-induced intestinal injury. MATERIAL/METHODS: Four groups of 10 1-day-old rat pups were studied. Hypoxia/reoxygenation (H/O)-induced intestinal injury was created. Group 1 was subjected to H/O just after birth and sacrificed at the end of the third day (Treatment Control). Group 2 was subjected to H/O just after birth and treated with sucralfate for 3 days. They were sacrificed at the end of the third day (Treatment). Group 3 was subjected to H/O on the third day after birth and then sacrificed (Prophylaxis Control). Group 4 was treated with sucralfate for the first 3 days, then H/O was created. Just after H/O, the pups were sacrificed (Prophylaxis). The intestinal tissues were harvested for histopathological investigation. Malondialdehyde (MDA) levels in the intestinal tissues were determined. RESULTS: The mucosal injury grades of the treatment and prophylaxis groups were significantly lower than those of control groups (p<0.05). The mean MDA level in the treatment and prophylaxis groups were 0.42+/-0.17 and 0.21+/-0.23 nmol/mg respectively. The MDA levels of both groups were significantly lower than in the control groups (p<0.05). CONCLUSIONS: The present study shows that sucralfate has beneficial effects in an experimental model of hypoxia/reoxygenation-induced intestinal injury. PMID- 15795692 TI - Chronic ethanol exposure does not impair urinary acidification even under stressful conditions. AB - BACKGROUND: Ethanol consumption is known to cause structural and functional renal damage in rat kidney. Acutely, ethanol impairs acid-base regulation, but a chronic effect on urine acidification has not been well elucidated. The aim of the present study was to assess urinary acidification in rats subjected to chronic ethanol consumption and renal function stressors, such as unilateral nephrectomy and/or metabolic acidosis. MATERIAL/METHODS: Ethanol-fed rats received aqueous ethanol 20% (v/v) as the sole drinking fluid for ten weeks. Then unilateral nephrectomy was performed and, five days later, metabolic acidosis was induced by administration of NH(4) Cl (1.5 mmol/kg body weight) twice a day for two days. The urinary acidification rate was assessed seven days after unilateral nephrectomy by determining urine pH, ammonium and titratable acid. RESULTS: Chronic ethanol consumption, with or without unilateral nephrectomy, did not change urine pH, titratable acid, or ammonium excretion. Although NH(4)Cl-induced metabolic acidosis resulted in a significant increase of the acidification rate, the changes were not significantly different between ethanol- fed and control groups. Renal (Na + K)-ATPase was enhanced by chronic ethanol consumption, as well as by unilateral nephrectomy, but not by NH(4)Cl-induced metabolic acidosis. CONCLUSIONS: Chronic ethanol consumption does not impair the urinary acidification function of the rat, even under stressful conditions, such as acid loading and/or unilateral nephrectomy. Although these data cannot be extrapolated to human settings, there is possible application when considering drinkers as potential kidney donors. PMID- 15795693 TI - Neuromuscular control of physiological tremor during elastic load. AB - BACKGROUND: Physiological tremor is an involuntary and continuous oscillation in every limb segment of a healthy human. This study investigates the neuromuscular control of physiological tremor by an elastic load applied to the middle finger. MATERIAL/METHODS: The subject maintained a stretching position of the middle finger against various tension springs. Physiological tremor was detected by an acceleration sensor attached to the middle finger. The motor-unit activity was estimated by surface electromyogram (EMG) measured from the extensor digitorum communis muscle with physiological tremor. The tremor and the EMG signals were analyzed by power and coherence spectra. RESULTS: Three frequency components appeared in physiological tremor while the subject extended the middle finger against the tension spring. They were classified as two load-independent components and a load-dependent component. Although the motor-unit activity was enhanced by the extension of the middle finger against a stiffer spring, the tremor amplitude did not significantly change. The extensor digitorum communis muscle produced a larger contraction force to maintain the finger posture against a stiffer spring, while the stiffness of the tension spring restrained the finger movement. The three frequency components of physiological tremor correlated with the motor-unit activity. These results supported the hypothesis of the origin of physiological tremor. The stretch-reflex system caused the load-dependent component, and the mechanical property of the elastic load determined its frequency. The supraspinal system produced the two load-independent components. CONCLUSIONS: The neuromuscular control to maintain the stretching of the middle finger against the elastic load gave rise to the three frequency components in physiological tremor. PMID- 15795694 TI - Impairment in orthostatic tolerance during heat exposure in individuals with Type I and Type II diabetes. AB - BACKGROUND: Diabetes is often associated with orthostatic hypotension and temperature intolerance. Studies have examined either one stress or the other and not both to see if the stresses are additive. MATERIAL/METHODS: Eight subjects with Type 1 and 12 subjects with Type 2 diabetes and 15 controls were tested for orthostatic intolerance during a vertical tilt in a thermally neutral environment (22 degrees C) and a warm room (42 degrees C). Blood pressure, cardiac output, stroke volume, heart rate, skin blood flow and skin and body temperature were recorded. RESULTS: All subjects experienced reductions in their cardiac output and blood pressure when tilted. In the warmer environment, significant impairments in the compensatory blood pressure response and redistribution of cardiac output were found in subjects with diabetes. When tilted to the vertical posture, subjects with diabetes experienced reductions of about 10 mmHg in systolic blood pressure in the cool environment, and a reduction of 21 mmHg when tilted in the warm environment. The fall in the systolic pressure in the subjects with diabetes was accompanied by corresponding reductions in stroke volume and cardiac output (from 5.06 L/min to 4.75 L/min) but no change in the heart rate. To compensate for a vertical tilt, skin blood flow was reduced by about half in the controls but there was only a small vasoconstriction in the subjects with diabetes. CONCLUSIONS: Individuals with Type 1 and Type 2 diabetes, while showing some loss in orthostatic tolerance in a cool environment, showed significant clinical impairment associated with heat exposure. PMID- 15795695 TI - A two-phased population epidemiological study of the safety of thimerosal containing vaccines: a follow-up analysis. AB - BACKGROUND: Thimerosal is an ethylmercury-containing preservative in vaccines. Toxicokinetic studies have shown children received doses of mercury from thimerosal-containing vaccines (TCVs) that were in excess of safety guidelines. Previously, an ecological study showing a significant association between TCVs and neurodevelopmental disorders (NDs) in the US was published in this journal. MATERIAL/METHODS: A two phased population-based epidemiological study was undertaken. Phase one evaluated reported NDs to the Vaccine Adverse Event Reporting System (VAERS) following thimerosal-containing Diphtheria-Tetanus acellular-Pertussis (DTaP) vaccines in comparison to thimerosal-free DTaP vaccines administered from 1997 through 2001. Phase two evaluated the automated Vaccine Safety Datalink (VSD) for cumulative exposures to mercury from TCVs at 1 , 2-, 3-, and 6-months-of-age for infants born from 1992 through 1997 and the eventual risk of developing NDs. RESULTS: Phase one showed significantly increased risks for autism, speech disorders, mental retardation, personality disorders, and thinking abnormalities reported to VAERS following thimerosal containing DTaP vaccines in comparison to thimerosal-free DTaP vaccines. Phase two showed significant associations between cumulative exposures to thimerosal and the following types of NDs: unspecified developmental delay, tics, attention deficit disorder (ADD), language delay, speech delay, and neurodevelopmental delays in general. CONCLUSIONS: This study showed that exposure to mercury from TCVs administered in the US was a consistent significant risk factor for the development of NDs. It is clear from these data and other recent publications linking TCVs with NDs that additional ND research should be undertaken in the context of evaluating mercury-associated exposures and thimerosal-free vaccines should be made available. PMID- 15795696 TI - Physical activity context and university student's propensity to meet the guidelines Centers for Disease Control and Prevention/American College of Sports Medicine. AB - BACKGROUND: Previous research shows that exercise context is important for exercise adherence--exercising alone is associated with reduced adherence whereas exercising with others is associated with increased adherence. The purpose of the study was to examine whether exercising in one or a combination of four contexts for physical activity (in a structured class, with others outside of a structured class, alone but in an exercise setting, and completely alone) is related to the degree to which university students meet prescribed (i.e., CDC/ACSM) guidelines for aerobic activity. MATERIAL/METHODS: Males (n=196) and females (n=398) completed a self-reported physical activity questionnaire pertaining to the frequency, intensity and duration of their activity in the four contexts outlined above. RESULTS: A positive relationship was found between the percentage of students meeting CDC/ACSM Guidelines and the number of contexts in which physical activity was undertaken. That is, a small percentage (9.9%) were active in a single context (i.e., only one context out of a possible four), with the majority of those (5.9%) engaging in physical activity with others outside of a structured setting. A larger percentage (28.9%) were active in two contexts, while 61.2% were active in three or more contexts. CONCLUSIONS: Health care professionals interested in motivating the physically inactive to become more active and the physically active to maintain activity at a frequency, intensity, and duration sufficient to meet the CDC/ACSM guidelines ought to promote opportunities for physical activity in a variety of social contexts. PMID- 15795697 TI - A comparative study of the role of erythropoietin in the pathogenesis of deficient erythropoiesis in idiopathic pulmonary fibrosis as opposed to chronic obstructive pulmonary disease. AB - BACKGROUND: Despite the severe derangement of gas exchange in the advanced stages of idiopathic pulmonary fibrosis (IPF), secondary erythrocytosis is either absent or much lower than is seen in chronic obstructive pulmonary disease (COPD) with comparable hypoxemia. This study investigates the differences in erythropoiesis between IPF and COPD, searching for the possible underlying mechanisms. MATERIAL/METHODS: The study included 32 patients with COPD, 18 patients with IPF, all with overt hypoxemia (PO(2) <65 mmHg), and 34 healthy controls. Erythrocytic parameters and serum erythropoietin (EPO) levels were assessed for all subjects. In a number of patients from both groups, the development of erythroid colonies grown from peripheral blood mononuclear cells was assayed in semisolid methylcellulose cultures and compared to cultures of control cells, in the presence of patient or control serum. RESULTS: Hb and serum EPO levels were significantly higher in the COPD group than in IPF patients and controls. However, the number of BFU-E colonies obtained from mononuclear cells of IPF patients was clearly higher than in COPD patients when the same culture medium was used. Unlike COPD sera, IPF sera induced a significant growth inhibition of erythroid bursts arising from mononuclear cells of both patients and controls. CONCLUSIONS: Our findings suggest a kind of ineffective erythropoiesis in IPF. Defective EPO production and inhibitory effect on erythropoiesis exerted by pro inflammatory cytokines released from alveolar macrophages may be implicated in the suboptimal erythropoietic response. However, the possible involvement of other factors affecting erythropoiesis in IPF requires further investigation. PMID- 15795699 TI - Variation in the duration of arousal in obstructive sleep apnea. AB - BACKGROUND: The aim of this study was to evaluate the relationship between the duration of arousal and the type, duration and minSaO(2) of apneas/hypopneas, and the time of night and sleep stage when they occurred. MATERIAL/METHODS: We measured the duration of arousals associated with apneas (A) and hypopneas (H) in 10 polysomnographic recording. Arousals were categorized as: 1) Short Electroencephalographic Arousal (SEA: 3-15 s) and 2) Awakenings (>15 s). The dependent variable for multiple logistic regression analysis was duration of arousal (0 = 3-11 s, 1 = >11 s); the independent variables were event (0 = H, 1 = A), event duration (0 = 10-20 s, 1 = >20 s), sleep stage (0 = REM, 1 = NREM 1-2); minSaO(2) (0 = 86%, 1 = <86%) and time of night (0 = at or before 3:00 AM, 1 = after 3:00 AM). RESULTS: We analyzed 1536 apneas and hypopneas. 80.3% had arousal, 61% SEA and 19.3% awakenings. The median duration of arousal was higher in apneas (A: 12 s, H: 9.2 s, p<0.001). The apneas and events >20 s with a minSaO(2) <86% had a higher probability of arousal >11 s (apnea: OR 2.1, p<0.001; events >20 s: OR 1.5, p<0.01; minSaO(2) <86%: OR 1.4, p<0.05). CONCLUSIONS: Arousals were greater in apneas than in hypopneas. Apneas and events >20 s with a minSaO(2) <86% were associated more frequently with arousal >11 s. PMID- 15795698 TI - Early development of immunity in diGeorge syndrome. AB - BACKGROUND: diGeorge syndrome is a relatively common congenital disorder with developmental defects, including hypoplasia or pathologic migration of the thymus, associated with deletion of contiguous genes on chromosome 22. We prospectively followed a cohort of children with confirmed 22q11.2 deletion. MATERIAL/METHODS: One to six repeated examination were performed in 13 boys and 21 girls, age 4 days to 19 years. Due to the proposed role of the thymus in T lymphocyte selection, we studied T lymphocytes and their function, and also the presence of double positive CD4+CD8+ and gamma/delta T lymphocytes in peripheral blood. RESULTS: A low number of T lymphocytes was detected in the majority of patients before the age of 2 years. Both spontaneous and PHA-induced proliferation were unexpectedly higher than in normal samples from children <2 years old. Both T cell numbers and function normalized thereafter in the majority of patients. Double positive T cells were detected in one boy, together with transient positivity of antinuclear antibodies. Gamma/delta T cells were greater than 5% in 21% of the children. In our 5-year prospective study we have not yet observed serious clinical signs of immunodeficiency or autoimmunity in these patients, except for repeated respiratory infections. CONCLUSIONS: All patients classified as partial diGeorge syndrome presented with delayed but gradual development of immune function against a background of impaired support by the thymus. PMID- 15795700 TI - Eradication rate of Helicobacter pylori in dyspeptic patients. AB - BACKGROUND: The eradication of Helicobacter pylori (HP) is dependent on several bacteriologic and geographic variables. Multiple-drug regimens have been tried in order to eradicate HPi; however, the optimal drug has not yet been found. MATERIAL/METHODS: In this single-blind, randomized clinical trial, 245 dyspeptic patients with positive rapid urease tests or positive cultures for HP were entered in the study and randomly treated with one of 3 drug regimens A, B, or C for 14 days. Each regimen was administered to 76 patients. Regimen A consisted of omeprazole 20 mg bid + bismuth subcitrate 120 mg 2 tab. bid + metronidazole 250 mg 2 tab. bid + tetracycline 250 mg 2 cap. bid. Regimen B consisted of omeprazole 20 mg bid + bismuth subcitrate 120 mg 2 tab. bid + furazolidone 100 mg 2 tab. bid + tetracycline 250 mg 2 cap. bid. Regimen C consisted of omeprazole 20 mg bid + bismuth subcitrate 120 mg 2 tab. bid + ciprofloxacin 500 mg bid. A urea breath test was performed 1-2 months after eradication. RESULTS: Of the 245 patients, 228 completed their treatment course. 57% were male and 43% female. Eradication rates were 76.3%, 68.4%, and 67.1% in groups A, B, and C, respectively (P=0.05). The eradication rate of drug regimen A was greater than B in males (86% vs. 63%; p<0.05), and that of B was greater than C in females (76.7 vs. 53.6%; P<0.05). CONCLUSIONS: The therapeutic regimen containing ciprofloxacin is preferable, although all drug regimens were less effective in Iran than in other parts of the world. PMID- 15795701 TI - Neurological complications secondary to severe scorpion envenomation. AB - BACKGROUND: The aim was to study the incidence, clinical manifestations, and prognosis of neurological complications secondary to scorpion envenomation. MATERIAL/METHODS: A retrospective study over a 13-year period including all patients admitted to our ICU for scorpion envenomation. RESULTS: During the period of study, 951 patients were admitted due to scorpion envenomation. Mean age was 14.7+/-17.4 years, ranging from 0.5 to 90 years. Neuromuscular signs were observed in 739 patients (78%), coma (Glasgow Coma Score 12) in 15.4% of cases, and convulsions in 6%. The other neurological signs observed were: agitation in 709 patients (74.6%), squint in 119 patients (12.5%), bilateral miosis in 43 patients (5%), and a bilateral mydriasis in 16 patients (1.7%). Brain CT was performed in 10 patients, these being abnormal in 90% of cases. In our study the presence of coma (p<0.001), convulsions (p<0.001), bilateral miosis (P<0.001), and the presence of bilateral mydriasis (P<0.001) correlated with poor outcome. CONCLUSIONS: Neurological manifestations were often observed in severe scorpion envenomed patients and they correlated with poor outcome. Their mechanisms are complex. Prevention is highly warranted. PMID- 15795702 TI - Relationships between erythrocyte membrane properties and components of metabolic syndrome in women. AB - BACKGROUND: Membrane structure in metabolic syndrome has not been fully investigated. The aim of our study was to determine the relationship between structural parameters of the erythrocyte membrane and blood pressure, indices of obesity, plasma lipids, and glucose levels in female patients with metabolic syndrome. MATERIAL/METHODS: In 23 women with metabolic syndrome and 12 control normotensive women, anthropometric indices, blood pressure, and serum lipids were evaluated and an oral glucose tolerance test was performed. Erythrocyte membranes obtained from these patients were investigated by electron spin resonance spectroscopy and the spin-labeling method. The order parameter S (membrane microviscosity), parameter h (index of membrane hydrophobicity), and the a/b ratio (index of the membrane surface configuration) were calculated. Data were analyzed using univariate correlation and stepwise multiple regression analysis. RESULTS: All membrane parameters differed significantly in women with metabolic syndrome from controls (P<0.001). The order parameter S was negatively related to waist circumference (R=-0.59, P<0.01) and positively to systolic blood pressure (R=0.44, P<0.05). Negative correlations of parameter h and the a/b ratio with postload plasma glucose level (R=-0.60 and -0.63, respectively, P<0.01) were observed. In stepwise multiple regression analysis the variation in the order parameter S was mainly explained by waist circumference and systolic BP (52%), and in parameter h and a/b ratio by postload glucose level (35% and 34%, respectively). CONCLUSIONS: Erythrocyte membrane structural changes in women are associated with features of metabolic syndrome, especially with abdominal obesity, high systolic blood pressure, and postload glucose level. PMID- 15795703 TI - Activity of enzymes with different subcellular localization in the blood plasma of patients with aortic aneurysm. AB - BACKGROUND: The wall of an abdominal aorta aneurysm shows increased activity of metalloproteinases, aspartyl cathepsins and cystein cathepsins, and decreased activity of inhibitors of these enzymes, as compared to the wall of a normal aorta. Parietal thrombi of the aneurysm also show marked proteolytic activity. The aim of our study was to determine the activity of secretory enzymes, membrane enzymes, cytosol enzymes, mitochondrial enzymes and lysosomal enzymes in the blood plasma of patients with abdominal aortic aneurysm. MATERIAL/METHODS: Blood for examinations was collected from 15 patients with abdominal aorta aneurysm without coexisting diseases, and from the same number of healthy subjects, to determine the activity of lactic acid dehydrogenase (LDH), 5' nucleotidase (5'NT), gamma-glutamyltranspeptidase (GGTP) and cholinesterase (ChE), alanine aminotransferase (ALT), aspartic aminotransferase (AST), cathepsin B (CB), cathepsin D (CD), and cathepsin L (CL). RESULTS: The plasma activity of ALT, AST and GGTP in the blood of patients with aortic aneurysm is lower than in the plasma of healthy subjects. The activity of LDH, 5'NT and ChE, CB and CL does not significantly differ in the plasma of patients with aortic aneurysm compared to the plasma of healthy subjects. The activity of CD is increased in the plasma of patients with aortic aneurysm. CONCLUSIONS: Determination of cathepsin D activity may be helpful in the diagnosis of aortic aneurysm. PMID- 15795704 TI - Echocardiographic assessment in minimally invasive mitral valve surgery. AB - BACKGROUND: Due to limited exposure, removal of intracavitary air and visual assessment of cardiac function during minimally invasive procedures are not always possible. We analysed the utility of intraoperative transesophageal echocardiography (TEE) and postoperative transthoracic echocardiographic (TTE) in minimally invasive mitral valve (MV) procedures. MATERIAL/METHODS: We evaluated data from 163 consecutive patients undergoing isolated minimally invasive MV replacement (n=40) or repair (n=123) via small right anterolateral thoracotomy (121 complex mitral procedures). Cardioplegic arrest was achieved using either endoaortic (n=23) or transthoracic aortic clamp (n=140). In addition to preoperative TTE, TEE was used intraoperatively before and after cardiopulmonary bypass (CPB). Postoperative TTE was performed to monitor valve function at 3 and 12 months, and at 5-year follow-up. RESULTS: Pre-CPB TEE was useful to assess valve dysfunction and assist in placement of the arterial and venous cannulas. During CPB, placement and positioning of the endoclamp were guided effectively in all but 4 patients, in whom recurrent balloon migration necessitated secondary transthoracic aortic clamping. TEE detected one acute retrograde aortic dissection and one circumflex artery occlusion. After 18.7+/-10.6 months follow up, all patients except three improved symptomatically and had consistently good valve function. CONCLUSIONS: Intraoperative TEE is essential for minimally invasive MV surgery, because it allows immediate control of valve function before and after surgery. It is useful to detect unexpected complications requiring immediate remedy. Postoperative echocardiographic results show that minimally invasive MV surgery is a good alternative to conventional surgery even in complex MV repairs. PMID- 15795705 TI - Randomized, controlled study of the effects of losartan versus enalapril in small doses on proteinuria and tubular injury in primary glomerulonephritis. AB - BACKGROUND: Pharmacological blockade of the renin-angiotensin-aldosteron system ameliorates glomerular and tubulointerstitial damage. For optimal renoprotection, high doses of angiotensin II converting enzyme inhibitors and angiotensin II subtype 1 receptor antagonists are commonly recommended, but cannot always be administered. The aim of this study was to evaluate the effects of low-dose (25 mg) losartan on proteinuria and tubular injury extent. MATERIAL/METHODS: This was an open, randomized, 12-month study on the effects of 25 mg of losartan (n=19) vs. 10 mg of enalapril (n=14) as a control on proteinuria, urinary excretion of N acetyl-beta-D-glucosaminidase (NAG), and blood pressure in patients with primary glomerulonephritis. The second part of the study was an uncontrolled assessment of the renal effects of 50-mg administration of losartan. RESULTS: There were no significant differences between the groups in the effects on proteinuria and NAG excretion. Losartan and enalapril reduced proteinuria by 32.8% (p<0.029) and 40.9% (p<0.021), respectively, but did not affect NAG excretion. The antiproteinuric effect of losartan, achieved without changes in blood pressure, was particularly evident in subjects with proteinuria >1.5 g/24 h and normal blood pressure. 50 mg of losartan caused a significant decrease in NAG excretion vs. the baseline (p<0.027). CONCLUSIONS: 25 mg of losartan and 10 mg of enalapril equally reduce proteinuria. The significant antiproteinuric effect of losartan was achieved despite no changes in blood pressure. There were no differences between the drugs regarding their influence on tubular injury extent. 50 mg of losartan seems to be the minimal dose to improve tubular status. PMID- 15795706 TI - Transplantation ethics from the Islamic point of view. AB - Organ transplantation has been transformed from an experimental procedure at Western academic centers to an increasingly common procedure in private and public hospitals throughout the world. Attendant with advancements in organ harvesting, preservation, and transplantation come moral issues. Islam is a holistic religion that takes into account social affairs of man as well as spiritual ones. Islam has a long history of ethics literature including the subgenre of medical ethics. Historical considerations are discussed as to why Muslim thinkers were late to consider contemporary medical issues such as organ donation. Islam respects life and values the needs of the living over the dead, thus allowing organ donation to be considered in certain circumstances. The sources of Islamic law are discussed in brief in order for non-Muslims to appreciate how the parameters of organ transplantation are derived. The Islamic viewpoint, both Shiite and Sunni, is examined in relation to organ donation and its various sources. The advantages and disadvantages of brain dead and cadaveric donation is reviewed with technical and ethical considerations. The Islamic concept of brain death, informed and proxy consent are also discussed. We discuss the concept of rewarded donation as a way to alleviate the current shortage of organs available for transplantation and consider secular and religious support for such a program. Suggestions are made for greater discussion and exchange of ideas between secular and religious thinkers in the Islamic world and between the Islamic world and secular Western countries. PMID- 15795707 TI - Vectors and delivery systems in gene therapy. AB - Gene therapy provides modern medicine with new perspectives that were unthinkable two decades ago. Progress in molecular biology and, especially, molecular medicine is now changing the basics of clinical medicine. Reports of long-term side effects of the first successful human gene therapy study have slowed the penetration of DNA usage into clinical routine. However, the main safety problem lies in the secure and efficient delivery of genes into target cells and tissues. A number of older and more recently discovered techniques have been developed for therapeutic gene transfer. A variety of viral and non-viral possibilities are available for basic and clinical research. This review summarizes the delivery routes and methods for gene transfer used in gene therapy. The best known are viral vectors derived from adenoviruses and adeno-associated viruses. Specific properties of lentiviruses and retroviruses ensure their status in current gene therapy procedures. Recent advances in the preparation of lipoplexes and greater safety support the use of naked DNA transfer. The latest approach, using bacteria as vectors for gene therapy or as in situ producers of therapeutic proteins (alternative gene therapy), must undergo critical evaluation in further experiments. PMID- 15795708 TI - Semicarbazide-sensitive amine oxidase (SSAO): from cell to circulation. AB - Semicarbazide-sensitive amine oxidase (SSAO) is a multi-functional enzyme widely present in nature. It converts primary amines into their corresponding aldehydes, while generating H(2)O(2) and NH(3). In mammals, SSAO circulates in plasma, while a membrane-bound form (often referred to as vascular adhesion protein-1, VAP-1) is found in many tissues and organs, especially in adipocytes and vascular endothelial and smooth muscle cells. In recent years, evidence has been accumulating that SSAO has a role in protein cross-linking, formation of advanced glycation end-products, atherogenesis, glucose regulation and leukocyte extravasation at inflammation sites. Plasma SSAO is quite stable in healthy adults, but is elevated in diabetes mellitus (both type 1 and type 2), congestive heart failure and liver cirrhosis. The origin of circulating SSAO remains unclear, but recent evidence from clinical studies and from (transgenic) animal studies suggests that adipocytes and vascular endothelial cells may be the most important source. Studies with cell cultures show evidence that the membrane bound SSAO can be split off from the cells, thus giving rise to the (truncated) circulating form of SSAO. In some pathological conditions the diseased organ may be the main source of the elevated plasma SSAO. Little is known as yet about the regulation of plasma SSAO. Thyroid hormone appears to play a (modest) role in this respect. Further evidence from clinical, animal and cell-culture studies, helped by the new availability of selective SSAO inhibitors, is needed to shed more light on the question of the regulation of SSAO. PMID- 15795709 TI - Genetic markers of restenosis after coronary angioplasty and after stent implantation. AB - Apart from coronary artery bypass grafting, percutaneous transluminal coronary angioplasty (PTCA) and intracoronary stent placement are well established treatment strategies for CAD. Substantial differences exist in the mechanisms of restenosis between conventional PTCA and stenting. Arterial remodeling is the main contributor to lumen re-narrowing after PTCA, whereas neointimal hyperplasia is almost the sole mechanism of restenosis after stenting. Several reports have demonstrated that genetic factors may be involved in the pathogenesis of restenosis after PTCA and in-stent restenosis. In this review the candidate genes involved in the pathogenesis of restenosis are analyzed as potential genetic markers of restenosis after PTCA and in-stent restenosis. The I/D angiotensin-I converting enzyme gene polymorphism, gene polymorphisms of the endothelial nitric oxide synthase (Glu298Asp, -786T>C), the glycoprotein IIIa PlA1/A2 gene polymorphism, gene polymorphism of the estrogen (PvuII), allele 2 of the interleukin-1ra gene, and the GT repeats in heme oxygenase-1 gene promoter may be used as genetic markers for in-stent-restenosis. On the other hand, only the stromelysin-1 5A/6A gene polymorphism and allele 2 of the interleukin-1ra gene may be used as a genetic marker for restenosis after PTCA. PMID- 15795710 TI - Testosterone and aggressiveness. AB - Aggressiveness is an ancestral behavior common to all animal species. Its neurophysiological mechanisms are similar in all vertebrates. Males are generally more aggressive than females. In this review, aggressive behavior in rodents, monkeys, and man and the role of testosterone and brain serotonin levels have been considered. Interspecific aggressiveness in rats has been studied considering the mouse-killing behavior; the neonatal androgenization of females increases adult mouse-killing as does the administration of testosterone in adults. Intraspecific aggressiveness was studied by putting two or more male rats (or mice) in the same cage; the condition of subjection or dominance is influenced by testosterone. In monkeys, testosterone is related to aggressiveness and dominance and, during the mating season, increases in testosterone levels and aggressive attitude are observed. In men, higher testosterone levels were obtained in perpetrators of violent crimes, in men from the army with antisocial behaviors, in subjects with impulsive behaviors, alcoholics and suicidals, in athletes using steroids, and during competitions. Aggressive and dominant behavior are distinguished. Testosterone influences both of these, even if man is usually inclined to affirm his power without causing physical damage. Testosterone receptors are mainly in some hypothalamic neurons, where it is aromatized into estrogens, which determine the increase in aggressiveness. A relation between testosterone levels and diencephalic serotonin has been shown: in fact, the lack of serotonin increases aggressive behaviors both in animals and man. Testosterone also increases ADH levels in the medial amygdala, lateral hypothalamus, and preoptical medial area, involved in aggressive behaviors. PMID- 15795711 TI - A randomized controlled trial of magnesium sulfate, in addition to usual care, for rate control in atrial fibrillation. AB - STUDY OBJECTIVES: We examine the safety and efficacy of magnesium sulfate infusion, in addition to usual care, for acute rate reduction in patients with atrial fibrillation and a rapid ventricular response rate. METHODS: This was a prospective, randomized, double-blind, placebo-controlled trial of intravenous magnesium sulfate in adult emergency department patients with rapid atrial fibrillation. Study solutions were given in addition to any therapy the treating physician would normally consider appropriate, including the use of standard rate reduction agents. Patients received either 20 mEq (2.5 g, 10 mmol) magnesium sulfate over a 20-minute period, followed by 20 mEq (2.5 g, 10 mmol) over a 2 hour period intravenously, or placebo. RESULTS: One hundred ninety-nine patients were randomized, 102 to receive magnesium sulfate and 97 to receive placebo. The antiarrhythmic drug most commonly used by treating physicians was digoxin. Magnesium sulfate was more likely than placebo to achieve a pulse rate of less than 100 beats/min (63 [65%] of 97 versus 32 [34%] of 93, relative risk [RR] 1.89; 95% confidence interval [CI] 1.38 to 2.59; P <.0001) and more likely to convert to sinus rhythm (25 [27%] of 94 patients versus 11 [12%] of 91 patients; RR 2.20; 95% CI 1.15 to 4.21; P =.01). Comparative mean pulse rate reductions in the magnesium sulfate group did not reach predetermined clinical significance levels (> or =15 beats/min reduction) at any of the measured time points. Magnesium sulfate was more likely to be associated with an adverse event (14 [15%] of 95 patients versus 5 [5%] of 92 patients; RR 2.71; 95% CI 1.02 to 7.23; P =.04). CONCLUSION: Magnesium sulfate, when used to supplement other standard rate-reduction therapies, enhances rate reduction and conversion to sinus rhythm in patients with rapid atrial fibrillation. PMID- 15795712 TI - Images in emergency medicine. Lightning strike. PMID- 15795713 TI - Influence of timing of troponin elevation on clinical outcomes and use of evidence-based therapies for patients with non-ST-segment elevation acute coronary syndromes. AB - STUDY OBJECTIVE: Previous studies in clinical trial populations have demonstrated that patients presenting with positive troponin levels have a higher risk of mortality than patients with later positive troponin levels, but the influence of the timing of troponin elevation has not been previously characterized. We evaluate the impact of the timing of troponin elevation on clinical outcomes and adherence to the American College of Cardiology/American Heart Association acute care guidelines for patients with non-ST-segment elevation acute coronary syndromes. METHODS: We examined inhospital outcomes and use of acute (<24 hours) medications and invasive cardiac procedures in 23,184 high-risk patients with non ST-segment elevation acute coronary syndromes (positive cardiac markers or ischemic ST-segment changes) during 2001 to 2002 from 396 US hospitals participating in the Can Rapid Risk Stratification of Unstable Angina Patients Suppress ADverse Outcomes with Early Implementation of the American College of Cardiology/American Heart Association Guidelines (CRUSADE) Initiative. Baseline and peak troponin values were recorded and were designated as positive if above the cutoff value used at each institution to designate definite myocardial necrosis. RESULTS: In the study cohort, 53.2% of patients presented with baseline positive troponin levels, 30.6% had baseline negative/later positive troponin levels, and 16.2% had negative troponin levels during the entire hospitalization. Patients with baseline positive troponin levels had a higher risk of inhospital mortality than patients with baseline negative/later positive troponin levels (6.5% versus 4.1%) and were less likely to undergo early cardiac catheterization or percutaneous coronary intervention. The use of acute aspirin (in approximately 91% of patients), heparin (85%), and beta-blockers (78%) was similar in patients with baseline-positive versus later-positive troponin levels. CONCLUSION: These findings demonstrate that evidence-based therapies and interventions for non-ST segment elevation acute coronary syndromes are underused in patients with elevated troponin levels, but baseline troponin elevations, which are associated with a higher risk of inhospital mortality, do not stimulate more aggressive care. PMID- 15795714 TI - 2004 American College of Cardiology/American Heart Association guidelines for the management of patients with ST-elevation myocardial infarction: implications for emergency department practice. AB - The American College of Cardiology and the American Heart Association last published evidence-based guidelines for the management of ST-segment elevation myocardial infarction (STEMI) in 1999. In mid-2004, in recognition of the evolution and improvement of many of the most basic tenets of clinical management of STEMI since that time, an updated edition of the STEMI guidelines has been published. These guidelines offer many evidence-based recommendations that are pertinent to the out-of-hospital and emergency department care of STEMI patients, including initial evaluation, risk stratification, stabilizing management, and the choice between pharmacologic and mechanical revascularization. These are presented and discussed here. PMID- 15795716 TI - Stroking the data: re-analysis of the NINDS trial. PMID- 15795715 TI - The impact of imbalances in baseline stroke severity on outcome in the National Institute of Neurological Disorders and Stroke Recombinant Tissue Plasminogen Activator Stroke Study. AB - STUDY OBJECTIVE: The National Institute of Neurological Disorders and Stroke (NINDS) Recombinant Tissue Plasminogen Activator (rtPA) Stroke Study demonstrated a clinically meaningful and statistically significant benefit of tissue plasminogen activator (tPA). Adjusting for the baseline National Institutes of Health (NIH) Stroke Scale, the benefit of tPA remained. However, other authors suggest that an imbalance in baseline stroke severity between the tPA and placebo groups confounded the results. Another issue that has been raised concerns a possible increase in early mortality for individuals given tPA. In post hoc subgroup analysis, we describe the effect of tPA across a spectrum of time from stroke onset to treatment and stroke severity subgroups. Stroke severity was based on the NIH Stroke Scale. We also compare early mortality (2-week and 30 day) in the tPA and placebo groups. METHODS: Using combined data from the 2 NINDS rtPA Stroke Study trials, we performed post hoc subgroup analyses of 3-month favorable outcome (defined by the NIH Stroke Scale, Barthel, Rankin, and Glasgow Outcome Scales). We categorized patients from the trials into onset to treatment (0 to 90 minutes, 91 to 180 minutes) by NIH Stroke Scale (< or =5, 6 to 20, >20) subgroups. Analyses were adjusted for all variables previously shown to be associated with favorable outcome at 3 months. We also compared early mortality within onset-to-treatment subgroups. RESULTS: For all the 12 specified onset-to treatment-NIH Stroke Scale subgroups, the adjusted odds ratio for a favorable 3 month outcome was greater than 1.0 and favored tPA. We detected no difference in mortality between patients treated with rtPA and those treated with placebo by 2 weeks posttreatment (rtPA=9%, placebo=13%; P =.49) or by 30 days (rtPA=11%, placebo=16%; P =.30). CONCLUSION: These are descriptive post hoc subgroup analyses. Using cut points defined in previous critiques of the NINDS trials, these analyses give results consistent with previous NINDS Study Group reports. Baseline NIH Stroke Scale imbalance does not account for the better outcome of rtPA-treated patients. PMID- 15795717 TI - Data and safety monitoring in clinical research: a National Institute of Neurologic Disorders and Stroke perspective. AB - The National Institute of Neurologic Disorders and Stroke supports a broad spectrum of research in the diagnosis and treatment of neurologic disease. Emergency medicine is increasingly involved in clinical research for patients with neurologic emergencies. Independent data and safety monitoring are critical components of clinical trials to ensure the protection of patients and the scientific integrity of the research. We review National Institute of Neurologic Disorders and Stroke principles of data and safety monitoring and provide examples to illustrate key concepts. PMID- 15795718 TI - Parenteral dihydroergotamine for acute migraine headache: a systematic review of the literature. AB - STUDY OBJECTIVE: Many therapies are used in the treatment of acute migraine headache, with little agreement on effectiveness. This systematic review is designed to determine the effectiveness of parenteral dihydroergotamine in reducing pain, nausea, and relapse for episodes of acute migraine in adults. METHODS: Randomized controlled trials were identified using MEDLINE, EMBASE, other computerized databases, hand searching, bibliographies, and contacts with industry and authors. Studies in which dihydroergotamine (alone or in combination with an antiemetic) was compared with placebo or any other common migraine therapy were considered. Relevance, inclusion, and study quality were assessed independently by 2 reviewers. RESULTS: From 281 potentially relevant abstracts, 11 studies met the inclusion criteria. Solitary dihydroergotamine use was compared to sumatriptan and phenothiazines in 3 studies; results failed to demonstrate a significant benefit of dihydroergotamine over these therapies. In 8 combination treatment studies, heterogeneity in study methodology prevented statistical pooling. However, dihydroergotamine administered with an antiemetic was as effective as or more effective than meperidine, valproate, or ketorolac across all pain, nausea, and relapse outcomes reported in all studies. CONCLUSION: This evidence suggests that dihydroergotamine is not as effective as sumatriptan or phenothiazines as a single agent for treatment of acute migraine headache; however, when administered with an antiemetic, dihydroergotamine appears to be as effective as opiates, ketorolac, or valproate. Given its nonnarcotic properties, parenteral dihydroergotamine combined with an antiemetic should be considered as effective initial therapy in clinical practice. PMID- 15795720 TI - Oral or intravenous N-acetylcysteine for acetaminophen poisoning? PMID- 15795719 TI - The Australasian Clinical Toxicology Investigators Collaboration randomized trial of different loading infusion rates of N-acetylcysteine. AB - STUDY OBJECTIVE: We determine whether the incidence of adverse events caused by intravenous N -acetylcysteine is significantly less when the initial dose is infused over a 60-minute period compared with the standard infusion period of 15 minutes. A secondary objective is to assess the efficacy of the 2 treatment arms. METHODS: This was a multicenter, randomized, prospective trial of patients who presented with acetaminophen poisoning and who were treated with N acetylcysteine and had no history of hypersensitivity to N-acetylcysteine. Patients were randomly assigned to receive the initial dose of N-acetylcysteine over a 15-minute or 60-minute period. Baseline signs and symptoms and adverse events were serially evaluated before and during administration of N acetylcysteine. Tests of liver injury and coagulation were collected at baseline and then at 12-hour intervals. RESULTS: The study was designed with an 80% power to detect a halving of the incidence of adverse events. Of 180 evaluable patients, 109 patients were randomized to the 15-minute group and 71 patients were randomized to the 60-minute group. The incidence of drug-related adverse events was 45% in the 15-minute group and 38% in the 60-minute group (95% confidence interval -8% to 22%). The study did not demonstrate a reduction of drug-related adverse outcomes with the 60-minute infusion. Incidence of maximum alanine aminotransferase levels indicating hepatotoxicity (serum level >1,000 IU/L) was 6.8% (5.6% for 15-minute, 8.7% for 60-minute). The difference did not attain statistical significance. CONCLUSION: This study did not demonstrate a reduction of drug-related adverse outcomes with the 60-minute infusion. The study also confirmed that early treatment with N -acetylcysteine (within 8 hours of ingestion) is more effective than later treatment. PMID- 15795721 TI - Case report: compartment syndrome after a suspected black widow spider bite. AB - Widow spider envenomations generally produce systemic neurologic syndromes without significant local injury. We report a patient who sustained a black widow spider bite to the left forearm and presented to the emergency department with rhabdomyolysis and compartment syndrome. We documented a decrease in symptoms and compartment pressure after administration of antivenom. No surgical intervention was performed. We believe this report to be the first documenting compartment syndrome associated with black widow spider bite. PMID- 15795722 TI - Delayed-onset heparin-induced thrombocytopenia. AB - Delayed-onset heparin-induced thrombocytopenia is a syndrome in which thrombocytopenia and thrombosis begin several days after heparin discontinuation. Delayed-onset heparin-induced thrombocytopenia is caused by immunoglobulin G antibodies that are reactive against the heparin-platelet factor 4 complex in the absence of circulating heparin. We describe 2 patients with delayed-onset heparin induced thrombocytopenia who presented to the emergency department. An 88-year old man and a 62-year-old man experienced thrombocytopenia and thrombosis 9 or more days after heparin cessation and demonstrated a further decrease in platelet count on reexposure to heparin. Delayed-onset heparin-induced thrombocytopenia should be included in the differential diagnosis of a patient with recent heparin exposure who presents with thrombosis or thrombocytopenia. PMID- 15795723 TI - A randomized controlled trial of an emergency department-based interactive computer program to prevent alcohol misuse among injured adolescents. AB - STUDY OBJECTIVE: To determine whether an emergency department (ED)-based laptop computer intervention reduces the normative age-related increase in alcohol misuse compared with standard of care. METHODS: This was a randomized controlled trial conducted from October 11, 1999, to April 14, 2001, in a community teaching hospital and university medical center. Subjects were aged 14 to 18 years and with a minor injury. Controls and intervention participants completed a computer based questionnaire. Intervention participants also completed a laptop-based interactive computer program to affect alcohol misuse. Main outcome measures were Alcohol Misuse Index (Amidx) and binge-drinking episodes. Follow-up occurred by telephone at 3 and 12 months. Analysis included repeated-measures analysis of variance (alpha=0.05; power 0.80; effect size 0.10). RESULTS: Three hundred twenty-nine participants were randomized to the intervention group, and 326 participants were randomized to the control group. Two hundred ninety-five (89.7%) intervention subjects and 285 (87.4%) control subjects completed 3- and 12-month follow-ups. For intervention and control groups, respectively, mean age was 16.0 and 15.9 years and men composed 66.8% and 66.3% of the groups; Amidx scores were 2.2 and 2.0; binge-drinking episodes were 1.2 and 1.0. Outcomes for intervention and control, respectively, were Amidx (3 months) 1.5 and 1.4; Amidx (12 months) 1.8 and 2.1; binge drinking (3 months) 0.9 and 0.8; and binge drinking (12 months) 1.4 and 1.2. Overall, there were no significant effects (effect size 0.04). No detrimental effects were noted. Subgroup analysis suggested that the intervention may have an effect among subjects with experience drinking and driving (5% of the sample). CONCLUSION: The intervention was not effective in decreasing alcohol misuse among the study population. Further research will be required to determine effectiveness among the subgroup of adolescent minor injury patients who have experience drinking and driving. PMID- 15795724 TI - Preventing alcohol misuse among adolescents. PMID- 15795725 TI - National Highway Traffic Safety Administration (NHTSA) notes. Drowsy driving. PMID- 15795727 TI - Suggestions for improving the reporting of clinical research: the role of narrative. AB - This article discusses the importance of narrative in reports of medical research. Stripped of all jargon and analytic technique, a scientific investigation is a story, and it is the nuances and details of the story that make it interpretable. While guidelines such as CONSORT have greatly improved the reporting of research, such guidelines are insufficient to ensure a meaningful reporting. The provision of explanatory narrative is essential. In this article, I propose that authors and journals exploit opportunities created by the worldwide Web to post supplementary material regarding their conception of the investigation, their execution of the study, their assumptions and limitations, and their rationale for any modeling efforts. I provide examples of how such narrative could be included in reports of randomized trials, observational studies, and studies of diagnostic tests. PMID- 15795728 TI - The quality of medical record review studies in the international emergency medicine literature. AB - STUDY OBJECTIVE: We assess the methodologic quality of studies using medical record review methodology in 4 international emergency medicine journals. A secondary aim was to compare methodology quality among these journals and across years. METHODS: This was an observational study of articles whose main methodology was medical record review published in Academic Emergency Medicine (AEM) , Annals of Emergency Medicine (Annals) , Emergency Medicine Journal (EMJ) , and Emergency Medicine Australasia (EMA) between January 2002 and May 2004. Eligible articles were reviewed for reporting of a clear hypothesis or objective, training of abstractors, defined inclusion and exclusion criteria, use of a standard abstraction form, definition of important variables, monitoring of abstractor performance, blinding of abstractors to study hypothesis, reporting of interrater reliability, sample size or power calculation, reporting of ethics approval or waiver, and disclosure of funding source. The primary outcome was the proportion of articles meeting each criterion. Secondary outcomes were comparison of the proportions of articles meeting each criterion among journals and by years. RESULTS: One hundred seven articles were analyzed; 31 were published in AEM, 29 in Annals, 29 in EMJ, and 18 in EMA . A clear aim was reported in 93% of articles, standardized abstraction forms were reported in 51%, interrater reliability was reported in 25%, ethics approval or waiver was reported in 68%, and sample size or power calculation was reported in 10%. CONCLUSION: Adherence to the quality criteria for medical record reviews was suboptimal, and there were significant differences among journals in overall methodologic quality. PMID- 15795729 TI - Reassessing the methods of medical record review studies in emergency medicine research. AB - STUDY OBJECTIVE: An assessment of the methods of medical record review studies published in emergency medicine journals during a 5-year period ending in 1993 provided strategies for improvements. We assess and quantify the current methodologic quality of medical record review studies in emergency medicine journals using published guidelines and compare these results among journals and with those of 10 years previously. METHODS: Independent, systematic searches of emergency medicine journals identified all medical record review studies published in 2003. Methodology assessments of each selected study were conducted independently by 2 other researchers, and disagreements were resolved by arbitration. RESULTS: We identified 79 (14%) medical record review studies in 563 original research articles in 6 emergency medicine journals. The highest adherence to methodologic standards was found for sampling method (99%; 95% confidence interval [CI] 93% to 100%), and the lowest was for abstractor blinding to hypothesis (4%; 95% CI 1% to 11%). Interobserver agreement for the 12 criteria ranged from 57% to 95%. A comparison of these results with those of 10 years ago revealed significant improvements in 3 of the 8 original criteria assessed: data abstraction forms, mentioning interobserver performance, and testing interobserver performance. CONCLUSION: Medical record review studies continue to comprise a substantial proportion of original research in the emergency medicine literature. Important improvements are noted in some criteria, but adherence remains below 50% for 7 of the 12 criteria assessed. PMID- 15795731 TI - Who does know? PMID- 15795730 TI - Medical record reviews in emergency medicine: the blessing and the curse. PMID- 15795732 TI - Images in emergency medicine. The hair tourniquet syndrome. PMID- 15795733 TI - Ultrasonography of the internal jugular vein in patients with dyspnea. PMID- 15795734 TI - Cruise ship medicine--2004. PMID- 15795736 TI - Tissue oxygen saturation monitoring in diagnosing necrotizing fasciitis of the lower limb: a valuable tool but only for a select few. PMID- 15795738 TI - Revolutionizing peer review? PMID- 15795740 TI - Photoreceptor axons play hide and seek. PMID- 15795741 TI - Illuminating the calcium sensor for exocytosis in a flash. PMID- 15795742 TI - Alzheimer disease, in living color. PMID- 15795743 TI - Olfactory coding: sniffing out signals. PMID- 15795744 TI - Touche: the feeling of choice. PMID- 15795748 TI - Obesity, macrophage migration inhibitory factor, and weight loss. AB - OBJECTIVE: Elevated macrophage migration inhibitory factor (MIF) has been implicated as a causal mechanism in a number of disease conditions including cardiovascular disease (CVD), diabetes, and cancer. Excess body fat is associated with an increased risk of numerous health conditions including CVD, diabetes, and cancer. To our knowledge, the association between MIF and obesity status and the effect of weight loss on serum MIF concentrations have not been reported. In this study, we examined the effects of participation in a behavior-based weight loss program on MIF concentrations in obese individuals. SUBJECTS: Study participants were 71 men and women enrolled in The Cooper Institute Weight Management Program. Participants were predominantly female (68%, n=48), middle-aged (46.5+/-9.8 y), and severely obese (BMI=43.0+/-8.6). METHOD: Plasma MIF concentrations and other standard risk factors were measured before and after participation in a diet and physical activity based weight management program. RESULTS: The mean follow-up was 8.5+/-3.0 months with an average weight loss of 14.4 kg (P<0.001). The majority of clinical risk factors significantly improved at follow-up. Median levels of plasma MIF concentration were significantly lower at follow-up (median [IQR]; 5.1[3.6-10.3]) compared to baseline (8.4 [4.3-48.8]; P=0.0005). The percentage of participants with plasma MIF concentration > or =19.5 mg/nl (highest tertile at baseline) decreased from 33.8 to 5.6% (P<0.001). Further, elevated baseline plasma MIF concentration was associated with markers of beta cell dysfunction and reductions in MIF were associated with improvements in beta cell function. CONCLUSIONS: Circulating MIF concentrations are elevated in obese but otherwise healthy individuals; however, this elevation in MIF is not uniform across individuals. In obese individuals with elevated circulating MIF concentrations, participation in physical activity and a dietary-focused weight management program resulted in substantial reduction in MIF. PMID- 15795749 TI - Obesity-related correlation between C-reactive protein and the calculated 10-y Framingham Coronary Heart Disease Risk Score. AB - OBJECTIVE: To clarify the role of body mass index (BMI, kg/m(2)) adjustment in predictive models for cardiovascular events that add high-sensitivity C-reactive protein (hs-CRP) to the 10-y Framingham Coronary Heart Disease Risk Score (FCRS). DESIGN: A cross-sectional study in a group of apparently healthy individuals. SUBJECTS: In all, 1512 apparently healthy individuals (955 men and 557 women) at a respective age of 49.7+/-10.6 and 50.6+/-9.6 y. RESULTS: The Pearson correlation between hs-CRP and the calculated 10-y FCRS was lower when adjusted for BMI. This reduction was especially noted in women where it dropped from 0.247 to 0.09. The dominant role of hs-CRP concentrations was also noted in a linear regression model, again, especially noted in women (drop of the standardized coefficient from 0.517 for BMI to 0.08 for the FCRS). CONCLUSIONS: The correlation between hs-CRP and the 10-y FCRS is partly related to the presence of obesity. We raise the possibility that the addition of BMI to the predictive model of FCRS might attenuate the cost-effectiveness of CRP measurements for this specific risk assessment. PMID- 15795750 TI - Antipsychotic drug-induced weight gain: development of an animal model. AB - OBJECTIVE: Weight gain is a prominent effect of most atypical antipsychotic drugs (AAPDs); yet, the mechanisms are not fully understood and no well-established mouse models exist for investigating the mechanisms. Thus, we developed a mouse model to evaluate the effects of AAPDs on eating, body weight (BW), and body composition. METHODS: Female C57BL/6J mice were used to test olanzapine, quetiapine, ziprasidone, and risperidone. Mice were acclimated to individual housing, given ad libitum access to chow and water, dosed with placebo peanut butter pills for 1 week, and then dosed daily with AAPD-laced peanut butter pills for 4 weeks. Weekly food intakes and BWs were measured, and body compositions were determined at the end of each experiment. RESULTS: After 4 weeks of treatment, olanzapine, quetiapine, ziprasidone, and risperidone caused significant weight increases, but only olanzapine and quetiapine were associated with significantly increased food intake. Body composition data revealed that olanzapine-treated mice had more relative fat mass and risperidone-treated mice had more relative lean mass than did control mice. Quetiapine and ziprasidone did not significantly affect relative body composition even though BW was increased. CONCLUSIONS: Oral AAPD administration causes increased BW in female mice. Our mouse model of AAPD-induced weight gain resembles the human response to these medications and will be used to investigate the mechanisms for weight gain and fat accumulation. PMID- 15795751 TI - Social ecological correlates of physical activity in normal weight, overweight, and obese individuals. AB - OBJECTIVE: Research is limited concerning the moderating influence of weight status (ie normal, over, and obese) on the social ecological correlates of physical activity (PA) in adults. Therefore, the present study attempted to shed light on this issue. DESIGN: In 2001, a national cross-sectional mail out panel survey was conducted over a 3-month period in the United States. SUBJECTS: There were 1867 normal weight (ie body mass index (BMI)=20-24.99 kg/m2), 2145 overweight (ie BMI=25-29.99 kg/m2), and 1902 obese (ie BMI>30 kg/m2) adults. MEASURES: Various demographic measurements were taken in addition to social support (SS), self-efficacy (SE), access to facilities, and PA. RESULTS: Normal weight individuals engaged in significantly more PA than overweight individuals, who in turn engaged in significantly more PA than obese individuals F(2,5991)=55.51, P<0.01. Further regression analyses showed that higher SE, SS, the access to facilities in a neighborhood, and various interactions among these constructs were significantly and positively associated with PA. Interestingly, the strength of these relationships varied depending on weight status. CONCLUSION: Weight status needs to be taken into consideration when examining social ecological correlates of PA. PMID- 15795752 TI - Anthropometric relationships between parents and children throughout childhood: the Fleurbaix-Laventie Ville Sante Study. AB - BACKGROUND: The study of parent-child anthropometric relationships and their evolution over time may help to better understand familial risk factors for childhood obesity. METHODS: In a population-based cohort of 124 nuclear families (Fleurbaix-Laventie Ville Sante Study (FLVS) I and II), various anthropometric parameters were measured in both parents and their children, first when the children were prepubescent and again at the end of puberty. Troncular adiposity repartition was estimated by calculating troncular to peripheral skinfolds ratio and waist-to-hip circumferences ratio. Birth and infancy heights and weights were also obtained from the children's health booklets. Parent-child correlations were estimated in infancy, before and at the end of the child's puberty. A prospective analysis was performed to predict the changes in the children's measurements over puberty according to their parents' corresponding baseline values. RESULTS: BMI and weight correlations at birth were high (>0.30) with the mother and low (<0.10) with the father, then they converged to an intermediate level at 2 y and remained between 0.2 and 0.3 thereafter. Correlations for waist circumference were already present at the prepubertal period and persisted on the same level at the postpubertal period, whereas correlations for subcutaneous adiposity - measured by four skinfolds - and for adiposity distribution - measured by ratios were higher at the postpubertal period. Moreover, the prospective approach showed that mother's BMI predicted the evolution of her children's BMI over puberty, whereas this relationship was observed more specifically with the father concerning adiposity distribution parameters. CONCLUSION: Maternal adiposity may act early in life on the adiposity of the child. Maternal and paternal adiposities seem to have quite distinct effects at two key periods of the child's adiposity development such as the prenatal and pubertal periods. PMID- 15795753 TI - Pursuing practical elegance in chemical synthesis. AB - In the 21st century, the field of chemistry will face more than just academic challenges. Indeed, our ability to devise straightforward and practical chemical syntheses is indispensable to the survival of our species. PMID- 15795754 TI - Supramolecular chemistry on water--towards self-assembling molecular electronic circuitry. AB - This feature article describes the self-assembly of electroactive molecules at the air/water interface, emphasizing the structural and electronic characterizations of the resulting supramolecular architectures. PMID- 15795755 TI - Hydrogen bond mediated open-frame networks in coordination polymers: supramolecular assemblies of Pr(III) and 3,5-dinitro-4-methylbenzoic acid with aza-donor compounds. AB - A coordination assembly of 3,5-dinitro-4-methylbenzoic acid and Pr(III), synthesized by hydrothermal methods forms a host structure, which is stable up to 300 [degree]C, through C-HO hydrogen bonds and accommodates different types of guest species varying from simple molecules like water to larger molecules like trans-1,2-bis(4-pyridyl)ethene. PMID- 15795756 TI - Synthesis of elliptical vanadoborates housing bimetallic centers [Zn4(B2O4H2)(V10B28O74H8)]8- and [Mn4(C2O4)(V10B28O74H8)]10-. AB - The hydrothermal synthesis of three new vanadoborate compounds with elliptical (V10B28O74H8) clusters is described. The clusters contain pairs of bimetallic Zn2 or Mn2 units. PMID- 15795757 TI - Fluorescent dendrimers with a peptide cathepsin B cleavage site for drug delivery applications. AB - The synthesis of a multifunctionally equipped first generation (G1) dendrimer carrying a pentapeptide with a cathepsin[space]B cleavage site, chelating ligands for Pt2+-complexation, and a dansyl fluorescence marker is described and an investigation of its cellular uptake as well as intracellular localization by confocal fluorescence microscopy reported. PMID- 15795758 TI - Micro-emulsion synthesis of monodisperse surface stabilized silicon nanocrystals. AB - Silicon nanocrystals with a uniform size distribution were synthesized in inverse micelles using powerful hydride reducing agents. The silicon nanocrystals surfaces were then stabilized with 1-heptene to produce particles with strong blue photoluminescence. PMID- 15795759 TI - X-ray crystallographic signature of supramolecular triple helix formation from a water soluble synthetic tetrapeptide. AB - Single crystal X-ray diffraction studies on the water soluble, synthetic tetrapeptide Tyr(1)-Aib(2)-Tyr(3)-Val(4) with a non-coded amino acid residue (Aib: [small alpha]-amino isobutyric acid) reveal that the peptide adopts an "S" shaped molecular structure which self-assembles to form a supramolecular triple helix using various non-covalent interactions including water mediated hydrogen bonds in the solid state. PMID- 15795760 TI - A tunable solid-state fluorescence system consisting of organic salts of anthracene-2,6-disulfonic acid with primary amines. AB - A dramatic change of solid-state fluorescence properties triggered by molecular arrangements of anthracene moieties was obtained by using organic salts of ADS with primary amines, indicating that modification of the amines enables us to tune the properties. PMID- 15795761 TI - Dynamic synthesis of a macrocycle containing a porphyrin and an electron donor. AB - New macrocycles incorporating a porphyrin and a [small pi] electron-rich aromatic were prepared from a dynamic disulfide library. The outcome could be influenced by use of templates. PMID- 15795762 TI - Water-assisted self-assembly of harmonic single and triple helices in a polymeric coordination complex. AB - A novel two-dimensional coordination polymer containing infinite, coherently pitched single and triple helical motifs is formed by the self-assembly of Cd2+, succinate, water and a bipyridyl ligand. PMID- 15795763 TI - Fluorescence enhancement by hydroperoxides based on a change in the intramolecular charge transfer character of benzofurazan. AB - Strong fluorescence signals were observed after the reaction of novel reagents with hydroperoxides. PMID- 15795764 TI - Aluminium(III) adsorption: a soft and simple method to prevent TiO2 deactivation during salicylic acid photodegradation. AB - Deposition of poisoning species on TiO2 during salicylic acid photodegradation can be halted when Al(III) has been previously adsorbed on the catalyst surface; this widens the application of photocatalysis to more concentrated solutions. PMID- 15795765 TI - Directed palladation: fine tuning permits the catalytic 2-alkenylation of indoles. AB - A C-H activating Pd-catalysed alkenylation of indole is regiospecific for 2 substitution when the nitrogen carries a 2-pyridylmethyl substituent. PMID- 15795766 TI - Polystyrenes with chiral phosphoramide substituents as Lewis base catalysts for asymmetric addition of allyltrichlorosilane: enhancement of catalytic performance by polymer effect. AB - In the asymmetric addition of allyltrichlorosilane to benzaldehyde, polystyrenes with chiral phosphoramide substituents as Lewis base catalysts showed up to 2.4 times better catalytic activity and 1.4 times higher enantioselectivity than the corresponding low-molecular-weight analogues. PMID- 15795768 TI - One-step process to fabricate Ag-polypyrrole coaxial nanocables. AB - Ag-polypyrrole nanocables were fabricated in aqueous solution at room temperature through a redox reaction between silver nitrite and pyrrole, using poly(vinyl pyrrolidone)(PVP) as assistant agent. PMID- 15795767 TI - Synthesis of a simplified analogue of eleutherobin via a Claisen rearrangement and ring closing metathesis strategy. AB - The enantioselective synthesis of a simplified eleutherobin analogue by ring closing metathesis (RCM) of the 2,9-divinyl-substituted tetrahydro-oxonin is described; the analogue and an advanced intermediate revealed microtubule stabilising properties in the micromolar range. PMID- 15795769 TI - Single-site heterogeneous Cr-based catalyst for the selective trimerisation of ethylene. AB - TAC-Cr3+/SiO2 complexes are highly active and selective ethylene trimerisation catalysts and possess single-site catalytic behaviour, an unusual property for heterogeneous catalysts. PMID- 15795770 TI - Controlling diastereoselectivity in the reactions of enantiomerically pure alpha bromoacyl-imidazolidinones with nitrogen nucleophiles: substitution reactions with retention or inversion of configuration. AB - Diastereoselective substitution reactions of [small alpha]-bromoacyl imidazolidinones with nitrogen nucleophiles can be promoted with either retention or inversion of configuration by carrying out reactions under epimerising or non epimerising conditions. PMID- 15795771 TI - Asymmetric amplification in asymmetric alternating copolymerization of cyclohexene oxide and carbon dioxide. AB - Asymmetric amplification in the copolymerization of cyclohexene oxide and carbon dioxide was demonstrated using chiral zinc complexes, prepared from diethylzinc, diphenyl(pyrrolidin-2-yl)methanol, and ethanol. PMID- 15795772 TI - Regioselective allylgallation of terminal alkynes. AB - The reactions of terminal alkynes with allylgallium reagents generated in situ from gallium and allyl bromides gave the corresponding 1,4-dienes in good yield via Markovnikov addition in THF at 70 degrees C. PMID- 15795773 TI - Fabrication of lotus-leaf-like nanoporous silica flakes with controlled thickness. AB - Lotus-leaf-like silica flakes with a three-dimensionally (3D) connected nanoporous structure and controllable thickness have been facilely synthesized; the flakes produced exhibited superior performance in adsorbing enzymes to their microspheric analogues. PMID- 15795774 TI - Synthesis of C2-chiral bifunctionalised spin labels and their application to troponin C. AB - An enantiomeric pair of C2-chiral bifunctionalised spin labels having a pyrrolidine nitroxide moiety, whose configurations were determined by X-ray crystal diffraction analysis, was prepared and applied to troponin C whose binding mode of double disulfide linkage was proved by EPR spectroscopy. PMID- 15795775 TI - Triol protection with 6-benzoyl-3,4-dihydro-(2H)-pyran. AB - 6-Benzoyl-3,4-dihydro-(2H)-pyran will protect 1,2,3-triols such as glycerol as their corresponding spiro-[5-phenyl-3,6,8-trioxabicyclo[3.2.1]octane-4,2[prime or minute]-tetrahydropyran]s and 1,2,4-triols (less efficiently) as the corresponding trioxabicyclo[3.2.2]nonanes; the hexol mannitol is converted into the corresponding bis-protected product. PMID- 15795776 TI - A solid-state dye-sensitized photovoltaic cell with a poly(N-vinyl-carbazole) hole transporter mediated by an alkali iodide. AB - A polymer-based dye-sensitized solar cell in dry solid state was constructed using poly(N-vinyl-carbazole)(PVK) as a hole-transporting layer; the cell attained the highest power efficiencies of 2.4% and 2.0% at 1/4 sun and 1 sun (AM1.5), respectively, by incorporation of a solid intermediate layer (LiI, KI) at the dye-PVK interface. PMID- 15795777 TI - Enhanced fluorescence and chiral discrimination for tartaric acid in a dual fluorophore boronic acid receptor. AB - The addition of D-tartaric acid to R,R- causes a large increase in fluorescence. While addition of L-tartaric acid to R,R- only produces small changes in fluorescence. PMID- 15795779 TI - Suppressed electron hopping in a Au nanoparticle/H2S system: development towards a H2S nanosensor. AB - We herein report and discuss electron transport within a Au/H2S nanoscale device and thereby highlight a phenomenon that may be used in the development of a novel on-chip H2S sensor. PMID- 15795778 TI - Synthesis and photophysics of a porphyrin-fullerene dyad assembled through Watson Crick hydrogen bonding. AB - A novel porphyrin-fullerene dyad assembled through Watson-Crick hydrogen bonds is described; this system undergoes photoinduced electron transfer upon irradiation with visible light to produce a charge separated state that is substantially longer lived than that of previous dyads of this type. PMID- 15795780 TI - Second-generation organocatalysts for the highly enantioselective dynamic kinetic resolution of azlactones. AB - Bifunctional organocatalysts of the thiourea-tert-amine type, carrying two "matched" elements of chirality, effect the alcoholytic dynamic kinetic resolution of a variety of azlactones with up to 95% ee. PMID- 15795781 TI - Highly chemoselective reduction of aromatic nitro compounds to the corresponding hydroxylamines catalysed by plant cells from a grape (Vitis vinifera L.). AB - Cells from a grape (Vitis vinifera L.) reduce aromatic nitro compounds under mild conditions to the corresponding hydroxylamines with unprecedented chemoselectivity. PMID- 15795782 TI - Biocatalytic racemisation of alpha-hydroxycarboxylic acids at physiological conditions. AB - Biocatalytic racemisation of aliphatic, aryl-aliphatic and aromatic alpha hydroxycarboxylic acids was accomplished using whole resting cells of Lactobacillus paracasei DSM 20207; the mild (physiological) reaction conditions ensured an essentially 'clean' isomerization in the absence of side reactions, such as elimination or decomposition. PMID- 15795783 TI - Novel chemoselective tosylation of the alcoholic hydroxyl group of syn-alpha,beta disubstituted beta-hydroxy carboxylic acids. AB - [small beta]-Hydroxy acids were readily converted into [small beta]-tosyloxy acids (hydroxyl group activation) in moderate to excellent yields via the O,O dianions generated by treatment with methyllithium and thus make it possible to prepare anti[small alpha],[small beta]-disubstituted [small beta]-lactones directly from the syn aldols. PMID- 15795784 TI - Anion directed synthesis of a hydrogensulfate selective luminescent rotaxane. AB - A new photo-active rhenium(I) bipyridyl based rotaxane has been synthesised which by virtue of the unique interlocked structural cavity selectively senses hydrogensulfate using luminescence spectroscopy. PMID- 15795785 TI - Encapsulation of metal particles within the wall structure of mesoporous carbons. AB - A method for the production of mesoporous carbon, with metal particles encapsulated in the walls to prevent leaching, is demonstrated by the synthesis of a cobalt containing CMK1 structure. PMID- 15795786 TI - Anhydrous solution synthesis of germanium nanocrystals from the germanium(II) precursor Ge[N(SiMe3)2]2. AB - A convenient, simple, single-source solution synthesis of Ge nanocrystals via thermal reduction of Ge(II) precursor Ge[N(SiMe3)2]2 in a non-coordinating solvent at 300 degrees C and 1 atm Ar is described. PMID- 15795787 TI - Fourteen-vertex homo- and heterobimetallic metallacarboranes. AB - Reduction of 4-(p-cymene)-4,1,12-closo-RuC2B10H12 followed by metallation with {M'} fragments (M' = {CpCo2+}, {(arene)Ru2+} or {(dppe)Ni2+}) affords 14-vertex bimetallic 1,14,2,10-RuM'C2B10 species having bicapped hexagonal antiprismatic structures. PMID- 15795788 TI - Novel poly(methimazolyl)borate complexes of niobium(V) and tantalum(V). AB - The reactions of [MCl4(eta-C5H5)](M = Nb, Ta) with Ph3Sn{HB(mt)3} (mt = methimazolyl) provide structurally characterised complexes of the new chlorobis(methimazolyl)borate ligand, [MCl3(eta-C5H5){kappa2-S,S'-HClB(mt)2}]. PMID- 15795789 TI - The substituted (S)-3-phenylpiperidine (-)-OSU6162 reduces apomorphine- and amphetamine-induced behaviour in Cebus apella monkeys. AB - Low affinity dopamine (DA) D2 antagonists such as the substituted (S)-3 phenylpiperidine (-)-OSU6162 have been proposed to be putative antipsychotic agents not endowed with extrapyramidal side effects (EPS). In the present study we investigated the effects of (-)-OSU6162 on (-)-apomorphine and d-amphetamine induced behaviours in EPS sensitised Cebus apella monkeys. (-)-OSU6162 was administered subcutaneously in doses of 1, 3, 6 and 9 mg/kg alone and in combination with (-)-apomorphine (0.25 mg/kg) or d-amphetamine (0.5 mg/kg). (-) OSU6162 inhibited (-)-apomorphine-(1-9 mg/kg) as well as d-amphetamine (3-9 mg/kg)-induced arousal and stereotypy. EPS did not occur when (-)-OSU6162 was administered in combination with (-)-apomorphine or d-amphetamine. However, when (-)-OSU6162 was administered alone, dystonia was observed at high doses (6 and 9 mg/kg) in two out of six monkeys. The present study shows that (-)-OSU6162 can inhibit (-)-apomorphine-induced behaviours in non-human primates at doses that do not cause EPS. When (-)-OSU6162 was tested against d-amphetamine-induced behaviours a separation between dose levels that inhibit d-amphetamine effects and cause EPS was not observed. The data further substantiate a role for low affinity DA D2 antagonists in the pharmacological treatment of psychosis. PMID- 15795790 TI - Polymorphism of the adrenergic receptor alpha 2a -1291C>G genetic variation and clozapine-induced weight gain. AB - Weight gain, leading to further morbidity and poor treatment compliance, is a common consequence of treatment with clozapine. The substantial interindividual and interracial differences in drug-induced weight gain suggest that genetic factors may be important. Several studies showed that alpha-2, adrenoceptor may related to feeding behavior with rat or lipolytic activity of human adipocyte tissue, they are related to body weight change. In the study, we try to test the possible relation of clozapine-induced weight gain and adrenergic receptor alpha 2a -1291C>G genetic polymorphism in a long term follow up (14.0 +/- 6.2 months). Our results show the genotype GG (8.45 +/- 7.2 Kg) with higher mean body weight gain than genotype CC (2.79 +/- 6.1 Kg) (p = 0.023). The finding identify a genetic factor associated with clozapine-induced weight gain in schizophrenic patients. PMID- 15795792 TI - Gastric emptying after semi-solid food in multiple system atrophy and Parkinson disease. AB - BACKGROUND: Gastrointestinal symptoms such as nausea, abdominal pain, and bloating are frequent complaints in patients with Parkinson disease (PD) and in patients with multiple system atrophy (MSA), and may be associated with delayed gastric emptying (GE). Although several GE studies in patients with PD have been performed, scant data exist in patients with MSA. METHODS: We assessed GE half times (T50) in 12 patients with MSA and compared them with those of 12 patients with PD and 12 age-matched healthy controls.GE was estimated scintigraphically using the left anterior oblique method after ingestion of a (99m)Tc colloid labeled balanced semi-solid meal (yogurt). GE data were obtained every 15 minutes until there was complete emptying of the stomach. Blood pressure, heart rate, plasma glucose and glucosylated hemoglobin were regularly determined. RESULTS: Reproducibility of the GE technique was excellent (Bland-Altman analysis, limits of agreement: -2.3 to 2.8). T50 was longer in MSA (82+/-3.4 min) and in PD (90.6+/-3.9 min) patients compared with controls (46.2+/-0.7) (two-way ANOVA, p<0.0001). T50 did not differ between patients with MSA and those with PD. No correlation existed between T50 and age, duration of the disease, magnitude of postprandial hypotension, levels of plasma glucose and glucosylated hemoglobin (Kendall's tau, p>0.05). CONCLUSIONS: Our results suggest that patients with MSA have GE rates similar to those of patients with PD, but slower than healthy age matched individuals. It remains to be investigated whether gastrointestinal dysfunction in MSA is related to both brain and peripheral pathology, as is presumed for PD. PMID- 15795791 TI - Creutzfeldt-Jakob disease mimicking corticobasal degeneration clinical and MRI data of a case. PMID- 15795793 TI - Hypoglycorrhachia in herpes zoster associated encephalitis of an immunocompetent young male: an unusual presentation. PMID- 15795794 TI - Intravascular electric impedance spectroscopy of atherosclerotic lesions using a new impedance catheter system. AB - Newer techniques are required to identify atherosclerotic lesions that are prone to rupture. Electric impedance spectroscopy (EIS) can characterize biological tissues by measuring the electrical impedance over a frequency range. We tested a newly designed intravascular impedance catheter (IC) by measuring the impedance of different stages of atherosclerosis induced in an animal rabbit model. Six female New Zealand White rabbits were fed for 17 weeks with a 5% cholesterol enriched diet to induce early forms of atherosclerotic plaques. All aortas were prepared from the aortic arch to the renal arteries and segments of 5-10 mm were marked by ink spots. A balloon catheter system with an integrated polyimide-based microelectrode structure was introduced into the aorta and the impedance was measured at each spot by using an impedance analyzer. The impedance was measured at frequencies of 1 kHz and 10 kHz and compared with the corresponding histomorphometric data of each aortic segment.Forty-four aortic segments without plaques and 48 segments with evolving atherosclerotic lesions could be exactly matched by the histomorphometric analysis. In normal aortic segments (P0) the change of the magnitude of impedance at 1 kHz and at 10 kHz (|Z|(1 kHz) - |Z|(10 kHz), = ICF) was 208.5 +/- 357.6 Omega. In the area of aortic segments with a plaque smaller than that of the aortic wall diameter (PI), the ICF was 137.7 +/- 192.8 Omega. (P 0 vs. P I; p = 0.52), whereas in aortic segments with plaque formations larger than the aortic wall (PII) the ICF was significantly lower 22.2 +/- 259.9 Omega. (P0 vs. PII; p = 0.002). Intravascular EIS could be successfully performed by using a newly designed microelectrode integrated onto a conventional coronary balloon catheter. In this experimental animal model atherosclerotic aortic lesions showed significantly higher ICF in comparison to the normal aortic tissue. PMID- 15795795 TI - Cardioprotection following adenosine kinase inhibition in rat hearts. AB - Adenosine kinase phosphorylates adenosine to AMP, the primary pathway for adenosine metabolism under basal conditions. Inhibition of adenosine kinase results in a site-specific increase in interstitial adenosine. Using a rat model of myocardial infarction, we examined the protective effects of adenosine kinase inhibition. Male Sprague-Dawley rats underwent 30 min regional occlusion followed by 90 min reperfusion. Infarct size, expressed as a percent of the area-at-risk, IS/AAR(%), was 58.0 +/- 2.1 % in untreated rats. Pretreatment with the adenosine kinase inhibitor, 5-iodotubercidin (1 mg/kg), limited infarct development to 37.5+/-3.7% (P < 0.001). The A(1) adenosine receptor (A(1)AR) antagonist, DPCPX (100 microg/kg), abolished the infarct-sparing effect of 5-iodotubercidin (IS, 62.8 +/- 1.3%). Similarly, the A(3) adenosine receptor (A(3)AR) antagonist, MRS 1523 (2 mg/kg), and the delta-opioid receptor (DOR) antagonist, BNTX, (1 mg/kg) abolished the reduction of IS produced by iodotubercidin. Pretreatment with the ROS scavenger, 2-MPG (20 mg/kg), or the PKC-delta antagonist, rottlerin (0.3 mg/kg) also abolished iodotubercidin-mediated cardioprotection. Furthermore, pretreatment with 5-HD, a mitochondrial K(ATP) (mitoK(ATP)) channel inhibitor, but not the sarcolemmal K(ATP) channel blocker, HMR-1098, abrogated the beneficial effects of adenosine kinase inhibition (IS, 59.5 +/- 3.8%). These data suggest that inhibition of adenosine kinase is effective in reducing infarct development via A(1)AR, A(3)AR and DOR activation. Data also suggest that this protection is mediated via ROS, PKC-delta and mitoK(ATP) channels. PMID- 15795796 TI - End-diastolic myofiber stress and ejection strain increase with ventricular volume overload--Serial in-vivo analyses in dogs with complete atrioventricular block. AB - BACKGROUND: Myocardial stress and strain are considered primary mechanical stimuli for hypertrophic remodeling. Their values and significance in the intact beating heart during chronic overload remain poorly characterized. METHODS AND RESULTS: Left-ventricular (LV) dimensions (echocardiography) and pressure (invasive) were simultaneously recorded in anesthetized dogs at sinus rhythm (SR), acute and 1, 2, 6, 12 weeks of atrioventricular block (AVB), leading to structural, electrical and contractile remodeling. Mechanical load of the myocardium was quantified as myofiber stress (sigma(f)), being force along myofiber orientation per cross-sectional area, and natural myofiber strain (e(f)), being change in natural logarithm of myofiber length (l) divided by its reference length: e(f) = ln(l/l(ref)). Time courses of sigma(f) and e(f) were calculated from LV pressure and dimensions, using a validated mathematical model of cardiac mechanics. End-diastolic sigmaf increased from 2.0 +/- 0.1 kPa at SR to 3.4 +/- 0.3 kPa at acute AVB, remaining elevated for > 6 weeks. Systolic sigma(f) was not affected by AVB. Ejection strain rose instantly upon AVB, reaching a maximum at 2 weeks: 0.24 +/- 0.02 vs. 0.10 +/- 0.01 at SR. The increase of myofiber stroke work (sigma(f)-e(f) loop area) from 3.1 +/- 0.3 at SR to 6.0 +/- 0.5 kJ/m(3)/beat at 1 week AVB was attributed mainly to an increase of strain during ejection. Stroke work and ejection strain remained elevated up to 12 weeks. The rate of LV-mass increase was maximal (2.2 +/- 0.4 g/day) at 1 week AVB. CONCLUSIONS: Serial mechanical phenotyping is feasible in the intact anesthetized dog with chronic ventricular overload. Our new approach yields values of mechanical load that are comparable to those found in isolated myocardium by others. In chronic AVB, both end-diastolic myofiber stress and ejection strain are increased. Early increases of both parameters coincide with peak hypertrophic growth, suggesting their important role for mechanotransduction. Peak systolic sigmaf is likely not important for hypertrophy in this model, since it does not change throughout the experiment. PMID- 15795797 TI - Design-based stereological estimation of the total number of cardiac myocytes in histological sections. AB - BACKGROUND: Counting the total number of cardiac myocytes has not previously been possible in ordinary histological sections using light microscopy (LM) due to difficulties in defining the myocyte borders properly. AIM: To describe a method by which the total number of cardiac myocytes is estimated in LM sections using design-based stereology. MATERIALS AND METHODS: From formalin-fixed left rat ventricles (LV) isotropic uniformly random sections were cut. The total number of myocyte nuclei per LV was estimated using the optical disector. Two-microm-thick serial paraffin sections were stained with antibodies against cadherin and type IV collagen to visualise the intercalated discs and the myocyte membranes, respectively. Using the physical disector in "local vertical windows" of the serial sections, the average number of nuclei per myocyte was estimated. RESULTS: The total number of myocyte nuclei in LV was 34.1 x 10(6) (0.08) (mean (coefficient of variation)), and the mean number of nuclei per myocyte 1.85 (0.03). Combining these estimates the total number of myocytes in LV was calculated to be 18.5 x 10(6) (0.09). CONCLUSIONS: This new method is applicable to a range of experiments focusing on myocyte proliferation and death. PMID- 15795798 TI - The new selective I(Ks)-blocking agent HMR 1556 restores sinus rhythm and prevents heart failure in pigs with persistent atrial fibrillation. AB - BACKGROUND: Antiarrhythmic drugs for treatment of atrial fibrillation in patients with heart failure are limited by proarrhythmia and low efficacy. Experimental studies indicate that the pure I(Ks) blocking agents chromanol 293b and HMR 1556 prolong repolarization more markedly at fast than at slow heart rates and during beta-adrenergic stimulation. These properties may overcome some of the above quoted limitations. METHODS AND RESULTS: Ten domestic swine underwent pacemaker implantation (PM) and atrial burst pacing to induce persistent AF. Four days after onset of persistent AF, pigs were randomized to HMR 1556 (30 mg/kg, p.o., 10 days) or placebo. All animals receiving HMR 1556 converted to SR (5.2 +/- 1.9 days), whereas placebo pigs remained in AF. Pigs treated with placebo developed high ventricular rates (297 +/- 5 bpm) and severe heart failure, whereas pigs treated with HMR 1556 remained hemodynamically stable. Left ventricular ejection fraction on the day of euthanization was significantly lower in the placebo compared to the HMR 1556 group (30 +/- 4% vs. 69 +/- 5%, p < 0.005). Similar results were seen with epinephrine levels (placebo 1563 +/- 193 pmol/l vs. HMR 613 +/-196 pmol/l, p < 0.05). Right atrial monophasic action potentials were significantly longer in the HMR 1556 compared to the placebo group (230 +/- 7 ms vs. 174 +/- 13 ms, p < 0.05). CONCLUSIONS: The new I(Ks) blocker HMR 1556 efficiently and safely restores SR and prevents CHF in a model of persistent AF. Restoration of SR is most likely linked to a marked prolongation of atrial repolarization even at high heart rates. PMID- 15795799 TI - "You cannot do nothing in this damn place": sex and intimacy among couples with an incarcerated male partner. AB - In an effort to deepen our understanding of how circumstances of forced separation and the interdiction of physical contact affect women's sexual behavior, we investigated the development and maintenance of heterosexual couples' intimacy when the male partner is incarcerated. As HIV-prevention scientists who work with women visiting men at a California state prison, we recognize that correctional control extends to these women's bodies, both when they are within the facility's walls visiting their mates and when they are at home striving to remain connected to absent men. This paper analyzes the impact of a peculiar public "place", a penitentiary, on couples' romantic and sexual interactions, drawing out the implications of imprisonment for relationship decision making, sexual health, and HIV risk. Using qualitative interviews with 20 women who visit their incarcerated partners and 13 correctional officers who interact with prison visitors, we examined how institutional constraints such as the regulation of women's apparel, the prohibition of physical contact, and the lack of forums for privacy result in couples forging alternative "spaces" in which their relationships occur. We describe how romantic scripts, the build-up of sexual tension during the incarceration period, and conditions of parole promote unprotected sexual intercourse and other HIV/STD risk behavior following release from prison. PMID- 15795800 TI - Experiencing release: sex environments and escapism for HIV-positive men who have sex with men. AB - There are nonsexual reasons that may motivate people to seek out sexual activity with others. Some men who have sex with men may seek out sex environments to engage in sexual behavior. Among the nonsexual reasons that exist for men who have sex with men is a desire to escape from distressing thoughts and feelings. The amplified sexuality and other unique characteristics of sex environments allow men to have more intense emotional experiences around sex. Using the cognitive escape model as a theoretical foundation, this analysis focuses on the emotional vulnerability that some of the men who visit these venues may be avoiding and how their experiences at these venues might act as releasing mechanisms to alleviate dissonant thoughts and feelings. Implications for public health services and future research are discussed. PMID- 15795801 TI - From nightlife conventions to daytime hidden agendas: dynamics of urban sexual territories in the South of France. AB - This article focuses on the space-time dynamics of sexual encounters in city neighborhoods associated with sex work and cruising. The data were collected as part of a multisite, longitudinal ethnographic study conducted in major cities of southern France. The results suggest that there is a continuous interaction between these sexualized public spaces and urban planning. Urban planners, however, are not the only actors shaping the location and the use of the space. Participants create a dialectical movement from night normative worlds to occult practices interwoven with daytime activities. They also create subtle social rules governing actors and their practices. PMID- 15795802 TI - Remembering the lizard: reconstructing sexuality in the rooms of narcotics anonymous. AB - The crack epidemic was devastating to poor American communities in part because of the destruction wrought by the system of exchanging sex for drugs, which was a key feature of the crack-use culture. Sex-for-drugs exchanges were often conducted under unsafe circumstances and were linked to the spread of AIDS and other STDs, as well as unplanned pregnancies. The existence of this alternative system of sexual relationships threatened the economic viability of established commercial sex work and undermined the status and power of women. Narcotics Anonymous (NA) meetings helped men and women recover from crack addiction through a well-described 12-step process. Described as the rooms, these time- and space specific encounters helped people become sober in the context of neighborhoods that were centers of the drug trade. Because of the key role of sex in the crack culture, transformation of sexual relationships was essential to establishing and maintaining sobriety. The manner in which the rooms of NA influence the sexuality and lifeworld of addicted people is explored using Barker's theory of ecological psychology. PMID- 15795804 TI - Making a place for escort work: a case study. AB - This case study focused on the steps taken by a Canadian city bordering the United States to manage the commercial sex component of an adult entertainment package that draws many American visitors. It explored how the city dealt with the potential of community stigmatization resulting from its growing reputation as a place for easily accessible sex trade. The distinctive Canadian laws related to the sex industry, the predominantly laissez-faire attitude toward people's sex lives, and concerns about maintaining the atmosphere of safety and privacy protection preferred by many residents set boundaries to the approach that could be taken. The specific historical, geographic, social, and political location of the city figured prominently in its choice of a normalization rather than a spatialization strategy. PMID- 15795803 TI - An appointment with god: AIDS, place, and spirituality. AB - This article describes how an African American gay man living with AIDS used his spiritual, religious, and cultural strengths to resist internalized dislocation because of heterosexism and homophobia. He was able to experience a relocation of God from places that rejected him to places that were conducive to his healing. By using these strengths, he was able to reject his physician's prediction of death and to call on God in response to an end-stage AIDS crisis. The development of spiritual agency is addressed. PMID- 15795805 TI - Big sister is watching you! Gender interaction and the unwritten rules of the Amsterdam red-light district. AB - It is generally accepted that in sexual interaction men are the hunters and women the gatherers. An expressly sexual environment such as a red-light district would be expected to only reinforce this image. Strikingly enough, however, it is precisely prostitutes who seem able to break with these established gender roles. This ethnographic study uses participant observation to demonstrate how assumptions about sexual interaction may be specious: female prostitutes are frequently not the passive victims and male clients not the active sex-hunters they are taken for. Both the communicative function and protection of privacy play an important role in these processes in a red-light district. In the red light district, gender interaction is bound by social regulation and unwritten rules that greatly exceed the influence of formal institutions. In this paper, the case of the red-light district "De Wallen" in Amsterdam, The Netherlands, is discussed in the context of several disciplines including urban anthropology, social psychology, geography, gender studies, and urban sociology. PMID- 15795806 TI - Your picture is your bait: use and meaning of cyberspace among gay men. AB - The Internet is seen by many as a form of cyberspace or environment in which to interact and socialise. This research project drew from the data of a quantitative and qualitative study of gay men in Perth, Western Australia. We examined gay men's usage patterns of chat rooms and other social aspects of the Internet to meet sexual partners. We then reviewed in detail the meanings gay men have for the various Internet environments, and the range of friendship, relationship, casual, or esoteric sex-seeking goals. We argue that gay men view and engage with the Internet differently from how they view and engage with other more traditional gay spaces. This different approach influences how interaction between the men, either online or face-to-face, progresses and how assumptions and expectations are built. This has implications for how sexual health promotion interventions determine the appropriate role and relationship they have with these online social spaces. PMID- 15795807 TI - "A whole stadium full": lesbian community at Women's National Basketball Association Games. AB - Using qualitative interviews with lesbian, bisexual, and heterosexual female Women's National Basketball Association (WNBA) fans, I argue that the (perceived) large lesbian attendance at WNBA games enables the construction of a lesbian community by lesbian and bisexual fans in a site qualitatively different from traditional locations of lesbian community. Because the WNBA is not an explicitly lesbian or queer space, the process of speculating about which players, coaches, and fans are lesbians is a meaningful part of being a WNBA fan for many lesbian and bisexual fans. Fantasizing a large lesbian presence enables lesbian and bisexual fans to actively and interpretively create lesbian community at the games. The women socialize and connect with lesbian and bisexual women, construct lesbian ownership of WNBA games, and define WNBA games as a unique space that is different from lesbian bars and other traditional "gay-only" spaces. PMID- 15795808 TI - Recruitment of the thigh muscles during sprint cycling by muscle functional magnetic resonance imaging. AB - The purpose of the present study was to investigate recruitment patterns of the thigh muscles during maximal sprint cycling by muscle functional magnetic resonance imaging (mfMRI). Twelve healthy men participated in this study and performed 2, 5, and 10 sets of 6-s supramaximal cycling with a load of 7.5 % of their body weight with 0.5 min of rest between the sets. Before and immediately after the exercise, T2-weighted MR images, i.e. mfMRI, of the right-thigh were taken to calculate T2 of eleven thigh muscles. Vastus lateralis, semitendinosus, and sartorius were the highest activated, i. e. had the greatest T2 change, among the quadriceps, hamstring, and adductors, respectively, compared with other muscles. Total power output during 2, 5, and 10 sets of sprint cycling was correlated with percent change in T2 in the quadriceps correlated (r (2) = 0.507 to 0.696, p < 0.01), the hamstring (r (2) = 0.162 to 0.335, p < 0.05 approximately 0.001), and the adductor muscles (r (2) = 0.162 to 0.473, p < 0.05 approximately 0.0001). With use of stepwise regression analysis, total power output was significantly correlated with % change in T2 of the vastus medialis (VM) (p < 0.0001) and vastus intermedius (VI) (p < 0.05) (r (2) = 0.698, p < 0.0001). We concluded that eleven thigh muscles were activated non-uniformly, and that the VM and VI play a key role during maximal sprint cycling. PMID- 15795809 TI - The aerobic fitness (VO2 peak) and alpha-fibrinogen genetic polymorphism in obese and non-obese Chinese boys. AB - The purpose of the study was to compare the aerobic fitness (VO (2) peak) between obese and non-obese boys at pre-puberty and examine the effect of body composition on VO (2) peak in this cohort with reference to TaqI polymorphism at alpha-fibrinogen gene locus. Seventy-seven Chinese boys with similar lifestyle participated in the study. Among them, 47 were diagnosed as obese. VO (2) peak was measured by a treadmill test and body composition was assessed via a combined anthropometrical and bioelectrical impedance analysis method. The alpha fibrinogen genetic polymorphism was detected through PCR-based digestion with TaqI restriction enzyme. The results indicated that VO (2) peak was significantly lower in obese boys compared with normal weight counterparts when the data were expressed either in conventional ratio unit (ml (-1) . min (-1) . lean body weight [LBW] (-1)) or in allometric unit (ml (-1) . min (-1) . body weight [BW] ( 2/3)). LBW, fat mass (FM), and body fat content (BF %) all were correlated with VO (2) peak, while LBW was the strongest predictor. The relationship between body composition and VO (2) peak seemed quite comparable across different alpha fibrinogen genotypes. Significant difference was observed between obese and non obese boys in terms of the proportion of genotypes and frequency of alleles. T1T1 homozygotes had higher risk for obesity. We came to the conclusion that prepubertal obese boys exhibited impaired aerobic fitness compared with their normal weight peers. VO (2) peak is closely related to LBW and independent of FM. This relationship remains constant irrespective of the TaqI alpha-fibrinogen genotypes that may be associated with fatness in boys. PMID- 15795810 TI - Oxygen uptake response to an 800-m running race. AB - We tested the hypothesis that time course of O (2) uptake (VO (2)) measured during a supramaximal exercise performed in the field is driven to maximal oxygen uptake (VO (2max)). On an outdoor track, five middle-distance male runners first performed a test to determine VO (2max) and a supramaximal 800-m running test at least two days apart. VO (2) response was measured from the start to the end of exercise with the use of a miniaturised telemetric gas exchange system (Cosmed K4). VO (2max) was reached by all subjects 45 +/- 11 s (mean +/- SD) after the onset of the 800-m race (i.e., 316 +/- 75 m), and was maintained during the next 33 +/- 6 s (i.e., 219 +/- 41 m). The mean relative exercise intensity of the 800 m was 120 % VO (2max). An unexpected significant decrease in VO (2) (24.1 +/- 7.0 %; p < 0.05) was observed in all subjects during the final 38 +/- 17 s (i.e., the last 265 +/- 104 m). We concluded that, at onset of a simulated 800 m running event, VO (2) is quickly projected towards the VO (2max), and then becomes limited by the achievable VO (2max). This race profile shown by all athletes is in some contrast to what can be expected from earlier findings in a laboratory setting. PMID- 15795811 TI - Gender differences in peak muscle performance during growth. AB - Gender-related differences in maximal leg muscle power were examined in 496 females and 426 males aged 8 to 20 years. Cycling peak power (CPP, including the force required to accelerate the flywheel of the cycle ergometer) was measured during three sprints. Optimal velocity (Vopt, velocity at CPP) was also determined. No gender-differences were observed in anthropometric characteristics and cycling performance between 8- and 14-year-old. From age 14, however, males showed a higher CPP than females, but also a higher lean leg volume (LLV, assessed by anthropometry). Allometric relationship between CPP and LLV (CPP = a . LLV ( b)) showed a clear gender-differentiation between 14- and 16-year-old: LLV exponent (b) was 1.05 in males vs. 0.74 in females. From 16 years onwards, analysis of covariance (ANCOVA) showed that the slopes of the CPP-LLV relationship were similar in both genders, but the intercepts differed. In other words, for a similar LLV, males showed greater CPP than females. It was suggested that this sex-related difference was due to total body fat increase, and more specifically lower-limb fat increase during puberty in girls, whilst the boys experienced increased lean body mass. Considering that the same gender-related difference was observed for optimal velocity adjusted for leg length, other factors such as fibre type variability or (and) neuromuscular activation might also be partly responsible for the higher peak muscle performance observed in males. PMID- 15795812 TI - The effect of two sports drinks and water on GI complaints and performance during an 18-km run. AB - Gastrointestinal (GI) complaints are frequently experienced during running. Sports drinks to prevent dehydration and hypoglycemia during exercise are generally used. The aim was to investigate the effect of 3 different drinks on GI complaints and performance during competitive running in a controlled field study. Ninety-eight well-trained subjects (90 M, 8 F, age 41 +/- 8 y) performed a competitive 18-km run three times within 8 days. The study was a controlled, standardized field experiment following a randomized, crossover design. Three different drinks were compared: water, a sports drink (CES), and a sports drink with added 150 mg/l caffeine (CAF). The incidence of GI complaints and the effect of the drinks on performance was studied. Each subject consumed 4 times 150 ml as follows: at the start, after 4.5 km, 9 km, and 13.5 km. Fluid intake was controlled. Incidence and intensity of GI complaints during the run were determined using a 10 points scale questionnaire. There were no significant differences in performance between the 3 drinks. Run time (18 km, mean +/- SD): WAT 1 : 18 : 03 +/- 08 : 30, CES 1 : 18 : 23 +/- 08 : 47, CAF 1 : 18 : 03 +/- 08 : 42. The use of carbohydrate-containing sports drinks led to higher incidences of all types of GI complaints compared to water. Significant differences (p < 0.05) were reached for flatulence; incidence: WAT 17.9 %, CES 28.6 %, CAF 30.6 %, and reflux; incidence: WAT 55.7 %, CES 78.6 %, CAF 72.5 %. There were no significant differences in intensity of the GI complaints. Addition of caffeine to CES had no effect on GI complaints, compared to CES alone. We conclude that sports drinks used during an 18-km run in cool environmental conditions do not support the performance better than mineral water. The use of sports drinks during an 18-km run leads to a higher incidence of both upper and lower GI complaints compared to water. Addition of caffeine to the sports drink has no effect on either running performance or GI complaints. PMID- 15795813 TI - The spatial-temporal and coordinative structures in elite male 100-m front crawl swimmers. AB - This study analysed the spatial-temporal and coordinative structures in 12 elite male 100-m front crawl swimmers. Swim performance was analysed over each length of a 25-m pool divided into five zones of 5 m. Velocity (V), stroke rate (SR), and stroke length (SL) were calculated for each zone and each length. Four stroke phases were identified by video analysis and the Index of Coordination (IdC) was established. Three modes of coordination were identified: catch-up (IdC < 0), opposition (IdC = 0), and superposition (IdC > 0). The swimmers tended to reduce the decrease in V and SR over the course of the 100 m by maintaining a stable SL. In fact, these spatial-temporal values were stable during the time spent stroking and were higher or lower during the start, the turns (in and out), and the finish. Thus the spatial-temporal changes did not occur within the lengths, but between them. Conversely, the evolution in the IdC showed that the swimmers had to install the stroke at the beginning and only reached a stable coordination in the second part of the race. Moreover, the IdC increased throughout the different zones of each 25-m length, indicating changes in motor organisation, particularly increases in the push or pull phases. The IdC values corresponded to a superposition of the arms, linked to a six-beat leg kick. Achievement of an effective superposition coordination occurred by boosting the stroke just after the turn-out until the end of the length. Regarding the spatial-temporal and coordinative structures of a 100-m front crawl, great swimming skill was reflected by both high and stable data. PMID- 15795814 TI - The validation of a new method that measures contact and flight times during vertical jump. AB - The aim of this study was to design and validate a vertical jump assessing system based on flight time measurements. Hence, the first phase of this study consisted of programming the software SportJumpv-1.0 and adapting a contact mat and a computer mouse in order to record the flight and contact times of a jump test. In the second phase, 9 subjects made 12 maximal and submaximal vertical single and rebound jumps. Thus 108 flight times and 36 contact times were simultaneously obtained with five different systems: ErgoJump Bosco System (PS), SportJump-v1.0 (SJ), Force Plate (FP), High-speed camera (HSC), and a Led (LED) filmed with a high-speed camera; FP was considered as the reference system. No significant differences were found either between SJ and LED, or between FP and HSC systems. SJ and LED overestimate flight time (9 ms and p < 0.001) and underestimate contact time in respect to FP values. PS underestimates flight time regarding FP values (4 ms and p < 0.01). In spite of the differences obtained between the systems, the errors found are constant. Those slightly depend on the subject's body mass for the systems that are connected to a contact mat. A high correlation between all of them was obtained (r approximately 1 and p < 0.001). The SJ system was validated for an accuracy of 500 Hz, and it was shown to be a useful tool in measuring flight and contact times during jumping. The differences and correlations found between the five systems show that it is necessary to take into account which system is being used when analyzing vertical jump performance. PMID- 15795815 TI - Successful treatment of osteitis pubis by using totally extraperitoneal endoscopic technique. AB - Osteitis pubis is characterized by pain, inflammation, and sclerosis in the pubic symphysis. It is often a self-limiting disease in athletes, but persistent pain may occasionally need surgery. Video-assisted placement of extraperitoneal retropubic synthetic mesh to support the damaged area may hasten the healing of this injury. During 1997 - 2002 five elite level male athletes with chronic groin pain associated with osteitis pubis were operated. The diagnosis was based on clinical findings, x-ray, magnetic resonance imaging (MRI), and isotope bone scanning. A 10 x 15 cm polypropylene mesh was placed into preperitoneal retropubic space using video-assisted technique. The pain and return to sport were asked at 1, 6, and 12 months after surgery. Preoperative technetium bone scan revealed an enhanced isotope uptake of pubic bone in each patient. T2 weighted MRI (n = 3) indicated bone marrow edema, which was decreased postoperatively on repeated MRI scans. Periosteal edema and irritation were also seen at operation. No complications were associated with the insertion of mesh. All 5 athletes returned to their sport activities between one to two months after surgery. After one year, no tenderness or pain was observed in the pubic bone. When conservative treatment fails, the placement of retropubic mesh is safe and a mini-invasive method to hasten the rehabilitation of osteitis pubis in selected cases. The postoperative recovery was uneventful, and the patients returned rapidly to their sporting activities. PMID- 15795816 TI - Few adverse effects of long-term creatine supplementation in a placebo-controlled trial. AB - Although oral creatine supplementation is very popular among athletes, no prospective placebo-controlled studies on the adverse effects of long-term supplementation have yet been conducted. We performed a double-blind, placebo controlled trial of creatine monohydrate in patients with the neurodegenerative disease amyotrophic lateral sclerosis, because of the neuroprotective effects it was shown to have in animal experiments. The purpose of this paper is to compare the adverse effects, and to describe the effects on indirect markers of renal function of long-term creatine supplementation. 175 subjects (age = 57.7 +/- 11.1 y) were randomly assigned to receive creatine monohydrate 10 g daily or placebo during an average period of 310 days. After one month, two months and from then on every fourth month, adverse effects were scored using dichotomous questionnaires, plasma urea concentrations were measured, and urinary creatine and albumin concentrations were determined. No significant differences in the occurrence at any time of adverse effects due to creatine supplementation were found (23 % nausea in the creatine group, vs. 24 % in the placebo group, 19 % gastro-intestinal discomfort in the creatine group, vs. 18 % in the placebo group, 35 % diarrhoea in the creatine group, vs. 24 % in the placebo group). After two months of treatment, oedematous limbs were seen more often in subjects using creatine, probably due to water retention. Severe diarrhoea (n = 2) and severe nausea (n = 1) caused 3 subjects in the creatine group to stop intake of creatine, after which these adverse effects subsided. Long-term supplementation of creatine did not lead to an increase of plasma urea levels (5.69 +/- 1.47 before treatment vs. 5.26 +/- 1.44 at the end of treatment) or to a higher prevalence of micro-albuminuria (5.4 % before treatment vs. 1.8 % at the end of treatment). PMID- 15795819 TI - Quality of life and psychomotor development after surgical treatment of hydrocephalus. AB - The purpose of this report is to analyse the long-term outcome in hydrocephalic children treated by shunt insertion, in particular their psycho-intellectual development and quality of life. We studied 46 patients aged 3 - 21, who had been operated on in the Department of Paediatric Surgery of the Medical University of Bialystok between 1982 - 2000 and had had ventriculo-peritoneal shunts inserted during their first year of life. Data from anamnesis and medical documentation were analysed. Age-appropriate psychomotor development and IQ tests were carried out: Wechsler Intelligence Scale for Children, Revised (WISC-R) (for children between the ages of 6 and 16), Wechsler Adult Intelligence Scale, Revised (WAIS-R PL) (for adults), Brunet-Lezine psychometric scale (early childhood), and Terman Merril intelligence scale (children younger than 3). The final IQs were above 90 in 30 % of children, between 70 and 90 in 24 %, between 50 and 70 in 26 %, and lower than 50 in 20 %. 69 % of patients presented with neurological deficits and visual or auditory deficits were found in 22 %. Integration into normal schools was possible for 58.7 % of the children, one of whom is now a second year medical student. A relationship between shunt malfunction and the children's development was observed. An essential aspect of caring for hydrocephalic children is their rehabilitation and integration into society. Early physical rehabilitation, stimulation of psychological development, and continued monitoring by a paediatric surgeon to ensure proper functioning of the shunts will improve the independence of such children in their families and among their peers. PMID- 15795817 TI - Cardiac mitochondrial respiratory function and oxidative stress: the role of exercise. AB - Investigations on the mechanisms capable of influencing heart mitochondrial function constitute a central contribution to the understanding of cardiac bioenergetics. In contrast to the conventional idea that reactive oxygen species (ROS) mostly act as a trigger for oxidative damage of biological structures, in low physiological concentrations they can regulate a variety of important molecular mechanisms, including those related to mitochondrial respiratory function. Among others, moderate physical exercise seems to be an important agent to induce cellular and mitochondrial environmental redox modifications and it is possible that these alterations could mediate cardiac mitochondrial respiration patterns. This brief review summarizes some current knowledge on mitochondrial respiratory pathways and focuses on data provided by studies dealing with exercise and cardiac respiratory mechanisms. It is emphasized the need of further experimental studies that analyze the association between physical exercise, particularly endurance training, and several mechanisms hypothetically related to the improvement of mitochondrial function, such as the overexpression of some important chaperone machinery and the up-regulation of both cellular and mitochondrial antioxidants. The influence of chronic moderate exercise on the functionality of some inner membrane components and on mitochondrial calcium loading capacity remains to be established. PMID- 15795820 TI - Results of Nuss procedures for the correction of pectus excavatum. AB - INTRODUCTION: The principle of the Nuss procedure is remodulation of the chest using a steel bar inserted by means of two small skin incisions along the axillary lines. PURPOSE OF THE STUDY: The assessment of the outcome after treatment. MATERIAL AND METHODS: Between July 1998 and March 2003 66 children were operated on by Nuss technique to correct pectus excavatum in our department. Age ranged from 1 to 19 years (mean 11.8 years). The evaluation prior to surgery included chest X-ray and CT, pulmonology consultation with pulmonary function tests and cardiology consultation. We used a modified technique in which we inserted the supporting bar directly from the right side, with no thoracoscope, no sharp tools and no guiding tape. Intrapleural or epidural bupivacaine was used for postoperative pain control. RESULTS: Duration of the operation was between 20 and 60 min. Complications included wound infections in six children, haemothorax in three and pneumonia in one. In 56 children the early cosmetic results were very good. In 5 children the excavation of the lower part of the sternum has persisted, in 5 children the excavation of the sternum was corrected but asymmetric thickening in the parasternal region persisted. In all cases the results were satisfactory for both the children and their parents. In 24 children the supporting bar was removed two years after the first operation. Postoperative courses were uneventful and the cosmetic results were good. In one child we observed a recurrence of the deformity six months after bar removal. CONCLUSIONS: The early results of the new operation for pectus excavatum are good in most children. The minimal invasiveness of this method suggests that this procedure is indicated in cases where the defects are only cosmetic. PMID- 15795821 TI - Interferon-alpha in the treatment of high-risk haemangiomas in infants. AB - BACKGROUND: Haemangiomas are benign tumours that may occasionally compress vital structures, or cause consumptive coagulopathy and heart failure. We describe our recent experience with interferon-alpha as a treatment modality for high-risk haemangiomas. PATIENTS AND METHODS: Eight children with high-risk haemangiomas were treated with interferon-alpha, six of which had failed previous steroid therapy. RESULT: Seven children responded to interferon therapy, one boy with a liver haemangioma died. Mild leucopenia and granulocytopenia were observed in all treated patients. Neurotoxicity occurred in 3 patients and was the most frequent serious complication. CONCLUSION: Interferon-alpha is an effective treatment modality for high-risk haemangiomas in children, especially in steroid-resistant patients. The most serious complication was early neurological toxicity. PMID- 15795822 TI - Evolution of diaphragmatic hernia management in the years 1991 - 2002. AB - AIM: Aim of the study was the analysis of our experience in the management of severe forms of CDH over the past 12 years. A comparison was made of pre- and postoperative strategy, with special attention focused on pharmacotherapy and respiratory therapy. METHODS: Between 1991 and 2002, a total of 43 newborns with CDH were treated, 24 (55.8 %) boys and 19 (44.2 %) girls. When evaluating the methods and results, the 12-year period was subdivided into two 6-year intervals. The correlation between mortality rates in particular groups was studied together with such parameters as gestational age, birth weight, hernial contents, the timing of surgery, persistent pulmonary hypertension and the type of preoperative management. RESULTS: A significant difference was noted in outcome in both groups. In the second 6-year period, the total mortality rate decreased from 59 % to less than 29 % compared to the first period (p < 0.05). Markedly higher mortality rates were seen in the subgroup of patients, in whom the hernia contained the stomach/liver tissue (62 % and 37 % in both respective periods). CONCLUSIONS: Most likely, the change in the outcome can be ascribed to the evolution of management in recent years. Thanks to a thorough preparation for surgery, aggressive pharmacotherapy and sophisticated methods of respiratory support, it has been possible to significantly decrease mortality rates in children with CDH. PMID- 15795823 TI - Active enlargement of the abdominal cavity--a new method for earlier closure of giant omphalocele and gastroschisis. AB - The authors present their own modification of surgical staged repair of large abdominal wall defects (AWD). 21 newborns with gastroschisis (GSCH) and 8 with omphalocele (OC) are presented. In all patients the protruding viscera were placed in a prosthetic sac with a semi-permeable lining. The sac was suspended using a system with active external traction with a force amounting to 30 - 40 % of the baby's weight. All babies tolerated the applied method well. A progressive stretching of the abdominal wall and enlargement of the abdominal cavity enabled the defect to be closed between the second and sixth postoperative day. In comparison to the classic method, the presented modification of staged repair of AWD makes an earlier reduction of the viscera into the abdomen possible and does not seem to be associated with a higher risk of mechanical or infectious complications. PMID- 15795824 TI - Uroflowmetry in children--is it necessary to repeat the study in a single patient? AB - Uroflowmetry, as the only non-invasive urodynamic means of assessing the function of the lower urinary tract is widely used in children. A possible psychological influence on the results is the reason why uroflowmetry is often repeated in the same patient. OBJECTIVES: The investigation was conducted to compare parameters of repeated uroflowmetry in a single child with the results of single uroflowmetry performed in a group of children. The goal of the study was to evaluate the efficacy of a single uroflow procedure in children. MATERIAL AND METHODS: A single uroflowmetric study was performed in 44 children aged from 9 - 11 years (mean 10 yrs). Out of the group one child was chosen who had 32 successive uroflow attempts. The range and average values of the uroflowmetric parameters were compared in both groups. Pearson's correlation coefficient was used for calculation and for a comparison of the uroflowmetric parameters of both groups (significance level p < 0.05). RESULTS: Average values of the voided volume, maximal and average flow were lower in the 32 attempts of a single child than in the control group (44 attempts). The values of the Pearson's correlation coefficient were ranged from 0.2 to 0.4 for flow/voided volume ratio and from 0.4 to 0.8 for flow time/voided volume ratio in both groups. There were no differences between the values in both groups apart from flow time/voided volume correlation (significance level p < 0.05). CONCLUSION: Repetition of uroflowmetric study does not seem to improve the efficacy of the modality in assessing voiding function in children. PMID- 15795826 TI - Tumour-like inflammatory abdominal conditions in children. AB - The presence of a tumour, poor general condition, features of anaemia, increased erythrocyte sedimentation rates and imaging suggesting malignancy were the common features in 4 different tumour-like abdominal conditions that are extremely rare in childhood. These conditions included: extensive retroperitoneal tumour with rib involvement that turned out to be an inflammatory lesion caused by Actinomyces in a 12-year-old girl; multi-loculated tumour of the mesentery/ovary caused by mesenteric lymphadenopathy in the course of a Salmonella enteritidis infection in a 2.5-year-old girl; tumour of the VII - VIII hepatic segments that turned out to be the focus of granuloma in the course of lambliasis in a 5.5-year old boy with a history of purulent neck lymphadenopathy and a final suspicion of immunocompromise; and a multi-loculated tumour of the small pelvis and inguinal area that turned out to be an abscess of the iliopsoas muscle in a 16-year-old boy. Apart from the imaging, the lesions required cytological examination of the material harvested by fine-needle biopsies (liver tumour) or histopathological investigations (retroperitoneal tumour, mesenteric/ovarian tumour, liver tumour and--on second surgery--the pelvic tumour) and/or bacteriological examination (all cases), serological examination (liver tumour and mesenteric/ovarian tumour), protozoal investigation (liver tumour), and measurement of AFP levels (mesenteric/ovarian tumour). Surgical treatment (retroperitoneal tumour, mesenteric/ovarian tumour and tumour of the small pelvis) and guided antibiotic therapy (all cases including 15 weeks of antibiotics in the first case) allowed complete recovery in 3 patients (actinomycosis, mesenteric lymphadenopathy, abscess of the iliopsoas muscle). Antibiotic and antiprotozoal therapy cured the granulomatous hepatitis; however this patient tended to develop severe right sided pleural/pulmonary changes (the child was referred for further diagnosis with suspicion of immunocompromise). PMID- 15795825 TI - Paediatric trauma in Spain: a report from the HUGM Trauma Registry. AB - OBJECTIVES: To analyse the characteristics of the infant population suffering trauma in our setting. To evaluate the importance of the different aetiological mechanisms. To study the pre- and intra-hospital management of these children. To describe the relative significance of the different lesions. To establish the magnitude of paediatric trauma as a social problem in terms of morbidity and mortality. MATERIAL AND METHODS: From January 1995 to April 2002, a total of 2633 children admitted to our Centre (Hospital Universitario Gregorio Maranon) after suffering some type of injury were included in our Trauma Register. 108 variables have been analysed, including the identification of the patient, type, site and mechanism of the accident, pre-hospital care, transport, complete evaluation on admission, indices of injury severity, diagnostic tests, lesions, treatments performed and morbidity and mortality. RESULTS: The accidents were more frequent in boys than in girls (68.5 % versus 31.5 %). The predominant age group was the 12 - 15 year old group (36.8 %). There was a higher frequency of accidents in the street (37.2 %) than at home (19.4 %) or at school (13.8 %). The most frequent mechanism was a fall (35.6 %), followed by road traffic accidents (23.7 %). On admission, 14.7 % of the children had a Paediatric Trauma Score (P.T.S.) < or = 8 (n = 388). 3.8 % were considered severe multiple trauma patients, presenting an Injury Severity Score (I.S.S.) > or = 15 (n = 101). 4.2 % of the children required intensive care. The most frequent lesions were those of the locomotor system (58.1 %) and head injuries (34.9 %). Some type of surgical or orthopaedic procedure was performed under general anaesthesia in 1522 patients (57.8 %). The mean length of stay was 4.4 days (range 1 - 214 days). Sequelae of some form were detected in 36.4 % of the patients over 3 years of age. The total mortality was 0.5 % (n = 13), being 12.8 % in the group of patients with an I.S.S. > or = 15. CONCLUSIONS: Analysis of the data in our Registry has helped us to define the characteristics of the paediatric trauma population in our setting, to monitor the management of trauma in the different care levels and to develop prevention programmes. It has also enabled us to compare the results with those of other centres in terms of morbidity and mortality with the aim of identifying and correcting any possible deficiencies in the care system. PMID- 15795827 TI - Haemorrhagic cystitis as a complication after allogenic bone marrow transplantation. AB - OBJECTIVE: The main objective of the following work is to present our own material and the ways in which we have dealt with haemorrhagic cystitis (HC) following allogenic bone marrow transplantation in children. MATERIALS AND METHOD: From 1994 to 2002, allogenic transplantation of haematopoietic cells was performed in 129 children at the Oncological and Haematological Child Clinic, Wroclaw University of Medicine. The procedure was carried out in patients with neoplastic diseases. In 33 cases, HC symptoms of various intensity were observed. The intensity of the symptoms was evaluated according to Arthur's four-point scale. To confirm the diagnosis USG was carried out in each case. Special attention was given to the ultrasonographic structure of the bladder wall. Cartoni's technique was followed in the examination. RESULTS: Out of 129 children who underwent allogenic transplantation of haematopoietic cells 33 (20.75 %) revealed HC symptoms. The symptoms occurred between the 2nd and the 124th day after transplantation (mean 29 days). The treatment included antiviral medicines, estrogens, reduction of immunosuppression and mechanical urological procedures. The children diagnosed with 2nd grade disease and higher were catheterised and diuresis was forced by the administration of larger amounts of liquids intravenously. Antihaemorrhagic drugs and vitamin K were also given. Blood was substituted if needed as were blood derivatives. Eighteen children with massive haematuria with clots underwent catherisation with a suprapubic catheter so as to continuously rinse the bladder. In 8 cases tamponade of the bladder occurred. The clots were removed from the bladder during cystoscopy under general anaesthesia. Twelve children died from HC. This amounts to 36 % of all the cases identified as HC and 9 % of all the children who underwent allogenic marrow transplantation. CONCLUSIONS: In conclusion it must be emphasised that HC in children after allogenic transplantation of haematopoietic cells is an extremely severe disease, which, if not cured, is terminal. The decision whether to perform embolisation of internal iliac arteries or to remove the bladder when non-radical methods have been exhausted, is worth considering. PMID- 15795828 TI - Malignant tumours in a horseshoe kidney in children: a diagnostic dilemma. AB - INTRODUCTION: Although malignant tumours in a horseshoe kidney are fairly common, both diagnosis and treatment are often difficult. The most frequently described tumour in children with this anomaly is nephroblastoma, but renal cell carcinoma and others can also occur. More than a hundred such cases have been presented in the English literature on the subject since Hildebrandt described nephroblastoma in a horseshoe kidney for the first time in 1895. Many of the articles report diagnostic and therapeutic problems. AIM: Our intention is to describe five more cases from our own experience. The cases involve malignant tumours in a horseshoe kidney. MATERIAL: Three children suffered from nephroblastoma, the other two had RCC. The tumours were particularly noted out of a total of 500 cases of nephroblastoma and 22 cases of other malignant tumours treated in Poland between 1993 and 2000. We also report here the treatment outcome of one case treated 30 years ago in our Clinic. CONCLUSION: Diagnostic and surgical procedures involved in the treatment of malignant tumours in the horseshoe kidney are complex and challenging. Only a few oncological centers cooperating with child urologists can provide comprehensive and effective treatment. PMID- 15795829 TI - Extrarenal Wilms tumour. AB - INTRODUCTION: Nephroblastoma is one of the most common solid tumours in children. It also is the most frequent tumour found in the kidneys. In 5 % of cases it affects both kidneys at the same time. About 70 - 80 new cases of Wilms tumour are registered in Poland annually, usually in patients aged from 1 to 7 years. Extrarenal Wilms tumours are extremely rare. Due to its rarity, series with more cases are based upon material collected from many clinical centers. AIM: We would like to present a case of a boy in whom we diagnosed nephroblastoma in the retroperitoneal space 14 years after he had completed a complex therapy for bilateral Wilms tumour. CONCLUSION: The development of an extrarenal tumour 14 years after complex treatment for bilateral nephroblastoma is related to the survival of metanephros located outside the kidney. PMID- 15795830 TI - Surgical treatment of congenital pseudarthrosis of the clavicle: a report of three cases and review of the literature. AB - Congenital pseudarthrosis of the clavicle is a rare entity of unknown aetiology with predominance of the right side. Our therapeutic concept is discussed with special reference to surgical therapy, histopathological findings and the most recent literature. Two girls and one boy, aged 4, 6, and 8 years, presenting with congenital pseudarthrosis of the clavicle were surgically treated between 1994 and 2000. A resection of the pseudarthrosis and internal fixation with a small reconstruction plate was performed. A bone graft from the iliac crest was used for restoration of clavicular length. Histological examination revealed a false joint with the ends of the clavicle covered by hyaline cartilage. The patients showed radiographic healing after 12 weeks. At follow-up (mean 44 months), all patients showed excellent clinical and radiological results without functional impairment. The clinical features and histological examination of the resected pseudarthroses clearly proved the diagnosis of a true congenital pseudarthrosis of the clavicle. According to our clinical and radiological results and considering the recent literature, we recommend surgical therapy with resection, bone grafting, and osteosynthesis with a reconstruction plate around the age of 4 - 6 years. PMID- 15795835 TI - Hospital infections in radiology clinics. AB - Hospital infections are serious concerns for health care workers and patients. Needle stick injuries, blood contacts, airborne infections and any kind of contamination pose a risk for hospital infections. The risk of hospital infection has been increased in radiology since the number of the patients and the exposure time between patients and radiology workers have increased especially with the usage of new modalities in the last three decades. Hospital infection risk and some universal standards and policies for protection were summarized in this article. PMID- 15795836 TI - Three-dimensional imaging and cone beam volume CT in C-arm angiography with flat panel detector. AB - We evaluated a new 3D angiography system with a flat panel detector (FPD) for its capabilitiy to acquire volume sets during a single rotation scan and to reconstruct high spatial resolution three-dimensional and cross sectional images, namely cone beam volume computed tomography (CBVCT) images. Present status of the technique, advantages and potential applications are discussed. PMID- 15795837 TI - Traumatic posterior fossa hematomas. AB - PURPOSE: Posterior fossa epidural hematomas are much less common than supratentorial epidural hematomas. The incidence of posterior fossa epidural hematomas among intracranial epidural hematomas has been reported to be 4% to 7%. Seven cases of posttraumatic posterior fossa epidural hematomas diagnosed by computed tomography (CT) are reported with radiological and clinical findings. MATERIALS AND METHODS: This study consisted of 7 posterior fossa epidural hematoma cases, out of 585 severe head trauma patients admitted and hospitalized in an 18-month period. The patients were evaluated regarding age, gender, type of trauma, cranial CT and Glasgow coma score in admittance, treatment and follow-up. RESULTS: Average age was 24.2 years and 85.7% of the cases were male. All cases had occipital fracture. Fifty-seven percent of the cases had only occipital fracture and posterior fossa epidural hematoma. All of the cases in this group were neurologically intact except for one who had a Glasgow coma score of 9 in admission. Two cases of this group were conservatively treated. In the others posterior fossa epidural hematomas got larger and they were treated surgically; these two cases recovered after surgery. Three of the 7 cases had the supratentorial region lesions; one of these cases died before operation. Two of them were treated surgically, one of them died and the other showed recovery after surgery. CONCLUSION: Acute posterior fossa epidural hematomas are usually symptom-free initially. After this silent period, clinical deterioration is quick to become fatal in most of patients. Surgery can be life-saving when performed in a timely manner. Therefore, CT should always be performed when an occipital trauma is diagnosed. PMID- 15795838 TI - Remote effect of optic tract trauma in the occipital lobe (case report). AB - Traumatic brain damage can result in severe visual impairments including hemianopia. Lesions correlated with hemianopia can be located in any part of the retrochiasmatic optic pathway. However, traumatic lesions of the optic tract are relatively rare. We present a case with posttraumatic left homonymous hemianopia who had signal intensity change at the ipsilateral optic tract on MR imaging and ipsilateral occipital hypoperfusion on SPECT imaging. PMID- 15795839 TI - Sarcomas: rare primary malignant tumors of the thorax. AB - In this article, it is aimed to review the radiological signs of unusual primary malignant tumors of the thorax. Radiological studies of 11 patients with histologic diagnosis of thoracic sarcomas were interpreted retrospectively. Tumors originated from the chest wall (n=3), mediastinum (n=4), and pulmonary parenchyma (n=4). Histopathologic diagnoses were fibrosarcoma (n=1), alveolar rhabdomyosarcoma (n=1), malignant hemangiopericytoma (n=1), malignant fibrous histiocytoma (n=2), pulmonary vein leiomyosarcoma (n=1), pulmonary artery sarcoma (n=2), pleuropulmonary blastoma (n=1), and chondrosarcoma (n=2). In order to evaluate thoracic sarcomas, cross-sectional methods such as CT and MRI can be useful in demonstrating the origin of the mass, relationship with and involvement of adjacent structures. They present as masses and, unfortunately, radiological findings are not sufficient for specific diagnosis. PMID- 15795840 TI - Multiple pulmonary metastases from intracranial meningioma: MR imaging findings (case report). AB - Extracranial metastases from meningioma are very rare and among those the lungs are the most common metastatic site. In the literature, pulmonary metastases of meningiomas have been emphasized on their rarity, non-specific imaging findings, and cytologic features. Some definitive MR imaging features of intracranial meningiomas have been described. However, MR imaging findings of the lung metastases of meningiomas have not been investigated with regard to these diagnostic imaging features. In this report, similarities between MR imaging features of primary intracranial meningioma and its rare lung metastases are presented. PMID- 15795841 TI - Diffuse pulmonary lymphangiomatosis: imaging findings. AB - Diffuse pulmonary lymphangiomatosis is a rare pulmonary disorder affecting the lymphatic channels from the mediastinum to the pleura. The disease usually occurs in children and young adults and frequently ends with death due to progressive course. Imaging findings of the disease are based on lymphatic involvement which appear as mediastinal soft tissue infiltration and thickening of pulmonary peribroncovascular bundles and interlobular septae. In this report, spiral and high-resolution computed tomography, and ultrasonography findings of severe form of this rare disease are presented. Furthermore, some lymphatic disorders, which are called with similar name but different appearances on imaging, are discussed. PMID- 15795842 TI - Preoperative evaluation of renal anatomy and renal masses with helical CT, 3D-CT and 3D-CT angiography. AB - PURPOSE: The aim of this prospective study was to determine the efficacy of three dimensional computed tomography (3D-CT) and three-dimensional computed tomographic angiography (3D-CTA) that were reconstructed by using the axial images of the multiphasic helical CT in the preoperative evaluation of renal masses and demonstration of renal anatomy. MATERIALS AND METHODS: Twenty patients that were suspected of having renal masses upon initial physical examination and ultrasonographic evaluation were examined through multiphasic helical CT. Two authors executed CT evaluations. Axial images were first examined and then used to reconstruct 3D-CT and 3D- CTA images. Number, location and size of the renal masses and other findings were noted. Renal vascularization and relationships of the renal masses with the neighboring renal structures were further investigated with 3D-CT and 3D-CTA images. RESULTS: Out of 20 patients, 13 had histopathologically proven renal cell carcinoma. The diagnoses of the remaining seven patients were xanthogranulomatous pyelonephritis, abscess, simple cyst, infected cyst, angiomyolipoma, oncocytoma and arteriovenous fistula. In the renal cell carcinoma group, 3 patients had stage I, 7 patients had stage II, and 3 patients had stage III disease. Sizes of renal cell carcinoma masses were between 23 mm to 60 mm (mean, 36 mm). Vascular invasion was shown in 2 renal cell carcinoma patients. Collecting system invasion was identified in 11 of 13 renal cell patients. These radiologic findings were confirmed with surgical specimens. CONCLUSION: Three-dimensional CT and 3D-CTA are non-invasive, effective imaging techniques for the preoperative evaluation of renal masses. PMID- 15795843 TI - Diagnostic value of ultrasonography in the evaluation of blunt abdominal trauma. AB - PURPOSE: To evaluate the diagnostic value of ultrasonography (US) in detecting intraabdominal injuries in patients with blunt abdominal trauma. MATERIALS AND METHODS: Blunt trauma patients admitted to the emergency department from January 2002 to August 2003 were retrospectively evaluated. A total of 454 patients with blunt abdominal trauma who underwent US examination were included. Ultrasonography results were compared with findings of CT, diagnostic peritoneal lavage, laparotomy and clinical course. Sensitivity, specificity, positive and negative predictive values of US in detecting free fluid, intraabdominal parenchymal organ injury or both were calculated. RESULTS: Computed tomography, diagnostic peritoneal lavage and laparotomy results showed that intraabdominal organ injury was present in 37 of 454 patients. Ultrasonography examinations were positive in 51 patients. True-positive findings were seen in 32 of these patients. In these 32 patients, US examination showed free fluid in 19, fluid and abdominal organ injury in 11 and only abdominal organ injury in 2. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy of US in detecting intraabdominal injury were 86.5%, 95.4%, 62.7%, 98.7% and 94.7%, respectively. CONCLUSION: Ultrasonography has high diagnostic performance in the screening of patients with blunt abdominal trauma. Abdominal US is a useful and valuable diagnostic tool after clinical evaluation in patients with blunt abdominal trauma. Because of its high negative predictive value, we recommend that clinical follow up is adequate for patients whose US results are negative for intraabdomial organ injury. PMID- 15795844 TI - Evaluation of the normal appendix at low-dose non-enhanced spiral CT. AB - PURPOSE: To evaluate the frequency of visualization, thickness and location of the normal appendix at non-enhanced spiral computed tomography (CT). MATERIALS AND METHODS: Low-dose spiral CT scans obtained for renal colic assessment in 243 patients were retrospectively reviewed. The frequency of visualization, thickness and location of normal appendices were recorded without knowledge of the patients' history for the appendectomy. RESULTS: Forty of 234 patients had a past history of appendectomy (17%). Sensitivity, specificity, positive and negative predictive values, and accuracy of visualization of the normal appendix were 71%, 85%, 96%, 37% and 73%, respectively. When no intraluminal content was visualized, this area was excluded from the measurement and the mean thickness was 3.4 mm+/ 0.66. CONCLUSION: The normal appendix is frequently seen at nonenhanced spiral CT. Knowing the normal thickness of the appendix can help reduce false positive and false negative diagnoses of acute appendicitis when reviewing non-enhanced spiral CT. PMID- 15795845 TI - Multislice CT angiography in the evaluation of hepatic vascular anatomy in potential right lobe donors. AB - PURPOSE: To assess the role of multislice CT angiography in the evaluation of arterial, hepatic and portal venous variations in potential right lobe donors. MATERIALS AND METHODS: Fifty-two potential liver donors (28 females, 24 males), underwent CT angiography in the arterial and portal venous phases with an eight row CT scanner. Two- and three-dimensional images were obtained from 1.25-mm thick axial images with multiplanar reformatting, maximum intensity projection and volume rendering techniques. Both axial and two- and threedimensional images were evaluated for possible variants of hepatic vasculature. In twelve operated patients, CT angiography results were compared with the results of surgery. RESULTS: Of 52 patients, 40 had type I, five type III, two type II, three type IX, and two type V hepatic arterial anatomy. In 13 patients (25%), segment IV artery originated from the right hepatic artery. In 26 patients (50%), veins that drained segment V and/or VIII to the middle hepatic vein were larger than 5 mm. Twenty-five patients (48%) had 28 accessory hepatic veins larger than 3 mm; 23 of these drained to the inferior vena cava more than 4 cm caudal to the right hepatic vein-vena cava junction. Three patients (6%) had trifurcation and one patient (2%) had quadrifurcation of the main portal vein. In two patients (4%), the right posterior portal vein arose directly from the main portal vein before its bifurcation. CT angiography findings showed one-to-one correlation with surgery in the 12 operated patients. CONCLUSION: Multislice CT angiography can successfully show the relevant hepatic vascular variations in potential liver donors. PMID- 15795846 TI - Testicular microlithiasis in pediatric age group: ultrasonography findings and literature review. AB - PURPOSE: To evaluate the occurrence of testicular microlithiasis in pediatric age group by means of ultrasonography (US) examinations and to review the literature for pediatric testicular microlithiasis cases accompanied by testicular and extratesticular tumors. MATERIALS AND METHODS: Nine children aged 3-16 years (mean age, 9.2 years) with testicular microlithiasis had been evaluated with US in a period ranging from 6 months to 6 years. In addition to the testicular ultrasonographic evaluation, liver US and abdominopelvic US were performed in all patients. RESULTS: Typical testicular microlithiasis findings were seen in a total of 17 testicles. In one patient, testis did not exist in either the scrotum or the inguinal canal or the abdomen unilaterally. None of the patients displayed a focal lesion during the evaluation. The abdominal ultrasonographic findings were normal in all patients. CONCLUSION: Although no tumoral lesion accompanying testicular microlithiasis or occurring in the course of evaluation was detected in this study, larger population and longer control periods are required, considering the co-existence of benign and malign lesions with testicular microlithiasis in the literature. PMID- 15795848 TI - Social structure, robustness, and policing cost in a cognitively sophisticated species. AB - Conflict management is one of the primary requirements for social complexity. Of the many forms of conflict management, one of the rarest and most interesting is third-party policing, or intervening impartially to control conflict. Third-party policing should be hard to evolve because policers personally pay a cost for intervening, while the benefits are diffused over the whole group. In this study we investigate the incidence and costs of policing in a primate society. We report quantitative evidence of non-kin policing in the nonhuman primate, the pigtailed macaque. We find that policing is effective at reducing the intensity of or terminating conflict when performed by the most powerful individuals. We define a measure, social power consensus, that predicts effective low-cost interventions by powerful individuals and ineffective, relatively costly interventions by low-power individuals. Finally, we develop a simple probabilistic model to explore whether the degree to which policing can effectively reduce the societal cost of conflict is dependent on variance in the distribution of power. Our data and simple model suggest that third-party policing effectiveness and cost are dependent on power structure and might emerge only in societies with high variance in power. PMID- 15795849 TI - A regulatory network analysis of phenotypic plasticity in yeast. AB - Models for the evolution of phenotypic plasticity suggest when and why plasticity might evolve. However, relatively little is known about the genetic basis of plasticity. Molecular studies have recently demonstrated that gene networks can provide a powerful way to infer phenotype from genotype. Information on the structure of the yeast gene regulatory network was combined with data on variation in gene expression in yeast across multiple environments in order to explore the genetic basis of phenotypic plasticity. The phenotypic plasticity of a gene was positively correlated with the number of transcription factors regulating that gene and was significantly lower for transcription factors than for downstream, nonregulatory genes. Plasticity of a gene was also affected by the local substructure of the network in which it was found and by the gene's function. These results illustrate how network analyses can be used to understand the complex genetic architecture of quantitative traits. PMID- 15795850 TI - On the theoretical and empirical framework for studying genetic interactions within and among species. AB - We present a quantitative genetic (QG) interpretation of the Bateson-Dobzhansky Muller (BDM) genetic model of speciation in order to unify the theoretical framework for understanding how the genetic differentiation of populations is associated with the process of speciation. Specifically, we compare the QG theory of joint scaling with the Turelli-Orr mathematical formulation of the BDM model. By formally linking the two models, we show that a wealth of empirical methods from QG can be brought to bear on the study of the genetic architecture of hybrid phenotypes to better understand the connections, if any, between microevolution within populations and macroevolution in the origin of species. By integrating the two theories, we make additional novel predictions that enrich the opportunities for empirically testing speciation genetic theory or facets of it, such as Haldane's rule. We show that the connection between the two theories is simple and straightforward for autosomal genes but not for sex-linked genes. Differences between the two approaches highlight key conceptual issues concerning the relevance of epistasis to evolution within and among lineages and to differences in the process of speciation in hermaphrodites and in organisms with separate sexes. PMID- 15795851 TI - Genetic basis and consequences of niche construction: plasticity-induced genetic constraints on the evolution of seed dispersal in Arabidopsis thaliana. AB - Because seed dispersal influences the environment experienced by seeds, that environment can change as dispersal evolves. The evolutionary potential of dispersal can in turn change as dispersal evolves, if its expression of genetic variation depends on the postdispersal environment. We examined whether seed dispersion patterns have a detectable genetic basis (and therefore evolutionary potential) and determined whether that genetic basis changed depending on one postdispersal environmental factor: conspecific density. We grew replicates of 12 ecotypes of Arabidopsis thaliana at high and low density and measured seed dispersion patterns and maternal traits associated with dispersal under controlled conditions. We found density-dependent ecotypic variation for maternal traits that influence dispersal. Significant genetic variation for postdispersal sibling density was detected only when plants were grown at high density, suggesting that if dispersal evolves to result in lower postdispersal densities, the expression of genetic variation for dispersal would be reduced. This dynamic could lead to a plasticity-induced constraint on the evolution of dispersal. The ability of organisms to alter the environment they experience and the ability of that environment to evolve can alter evolutionary dynamics by augmenting or reducing evolutionary potential and thereby facilitating or constraining evolutionary responses to selection. PMID- 15795852 TI - A dimensionless invariant for relative size at sex change in animals: explanation and implications. AB - Recent comparative studies across sex-changing animals have found that the relative size and age at sex change are strikingly invariant. In particular, 91% 97% of the variation in size at sex change across species can be explained by the simple rule that individuals change sex when they reach 72% of their maximum body size. However, this degree of invariance is surprising and has proved controversial. In particular, it is not clear why this result should hold, given that there is considerable biological variation across species in factors that can influence the evolutionarily stable timing of sex change. Our overall aim here is to explain this result and determine the implications for other life history variables. Specifically, we use a combination of approaches to formalize and make explicit previous analytical theory in this area, examine the robustness of the empirical invariance result, and carry out sensitivity analyses to determine what the empirical data imply about the mean value and variation in several key life-history variables. PMID- 15795853 TI - Female polymorphism, frequency dependence, and rapid evolutionary dynamics in natural populations. AB - Rapid evolutionary change over a few generations has been documented in natural populations. Such changes are observed as organisms invade new environments, and they are often triggered by changed interspecific interactions, such as differences in predation regimes. However, in spite of increased recognition of antagonistic male-female mating interactions, there is very limited evidence that such intraspecific interactions could cause rapid evolutionary dynamics in nature. This is because ecological and longitudinal data from natural populations have been lacking. Here we show that in a color-polymorphic damselfly species, male-female mating interactions lead to rapid evolutionary change in morph frequencies between generations. Field data and computer simulations indicate that these changes are driven by sexual conflict, in which morph fecundities are negatively affected by frequency- and density-dependent male mating harassment. These frequency-dependent processes prevent population divergence by maintaining a female polymorphism in most populations. Although these results contrast with the traditional view of how sexual conflict enhances the rate of population divergence, they are consistent with a recent theoretical model of how females may form discrete genetic clusters in response to male mating harassment. PMID- 15795854 TI - Maintenance of sex-linked deleterious alleles by selfing and group selection in metapopulations of the phytopathogenic fungus Microbotryum violaceum. AB - Microbotryum violaceum is a fungus that causes the sterilizing anther smut disease in many Caryophyllaceae. Its diploid teliospores are heterozygous at the mating-type locus, normally producing equal proportions of haploid sporidia of the two mating types. However, natural populations contain high frequencies of individuals producing sporidia of only one mating type. This mating-type ratio bias is caused by the presence of deleterious alleles at haploid phase ("haplo lethals") linked to the mating-type locus. These haplo-lethals can be transmitted if there is conjugation among the products of meiosis (intratetrad selfing). Haplo-lethals still suffer from selective disadvantages, through reducing the infection probability of strains that carry them, and thus cannot persist in a panmictic population. We develop a realistic model of a metapopulation of M. violaceum on its host Silene latifolia. Simulations show that if intratetrad selfing rate is high, haplo-lethals can be maintained under a metapopulation structure because of founder effects and selection at the population level. Populations founded only by strains carrying haplo-lethals experience a lower extinction rate precisely because of their lower infection ability; they spread more slowly and sterilize fewer plants, thereby allowing their host population to grow more rapidly and therefore to be less prone to extinction. PMID- 15795855 TI - Initial diversification of living amphibians predated the breakup of Pangaea. AB - The origin and divergence of the three living orders of amphibians (Anura, Caudata, Gymnophiona) and their main lineages are one of the most hotly debated topics in vertebrate evolution. Here, we present a robust molecular phylogeny based on the nuclear RAG1 gene as well as results from a variety of alternative independent molecular clock calibrations. Our analyses suggest that the origin and early divergence of the three living amphibian orders dates back to the Palaeozoic or early Mesozoic, before the breakup of Pangaea, and soon after the divergence from lobe-finned fishes. The resulting new biogeographic scenario, age estimate, and the inferred rapid divergence of the three lissamphibian orders may account for the lack of fossils that represent plausible ancestors or immediate sister taxa of all three orders and the heretofore paradoxical distribution of some amphibian fossil taxa. Furthermore, the ancient and rapid radiation of the three lissamphibian orders likely explains why branch lengths connecting their early nodes are particularly short, thus rendering phylogenetic inference of implicated relationships especially difficult. PMID- 15795856 TI - Correlates of species richness in mammals: body size, life history, and ecology. AB - We present the most extensive examination to date of proposed correlates of species richness. We use rigorous phylogenetic comparative techniques, data for 1,692 mammal species in four clades, and multivariate statistics to test four hypotheses about species richness and compare the evidence for each. Overall, we find strong support for the life-history model of diversification. Species richness is significantly correlated with shorter gestation period in the carnivores and large litter size in marsupials. These traits and short interbirth intervals are also associated with species richness in a pooled analysis of all four clades. Additionally, we find some support for the abundance hypotheses in different clades of mammals: abundance correlates positively with species richness in primates but negatively in microchiropterans. Our analyses provide no evidence that mammalian species richness is associated with body size or degree of sexual dimorphism. PMID- 15795858 TI - Evolutionary conflicts of interest: are female sexual decisions different? AB - Analyses of reproductive conflicts of interests have yielded important evolutionary insights in many areas of biology. The usefulness of conflict analyses of traits that have been traditionally interpreted as resulting from female choice is controversial, however. This article explores a possible explanation for why conflicts of interest may be ameliorated in female choice situations. In contrast to most other evolutionary contexts in which conflicts of interest are thought to have been important, sexual reproduction usually involves an extensive, irretrievable mixing of the genomes of the participants. Under certain combinations of costs and benefits to females, the genes in the female's genome can benefit, through increased reproduction of her offspring, from the very genes that produce sexually antagonistic traits in the male. In short, females can sometimes gain by "losing." Such Fisherian payoffs are also possible, though probably less important, for males. Gaining by losing is not feasible in most other contexts of evolutionary conflict, except under some conditions in parent-offspring conflict. Some apparent parent-offspring conflicts may instead be parental choice among offspring that is analogous to traditional female choice. Parent-offspring conflict may be relatively common, however, because offspring manipulation of their parents is likely to be damaging to the parents. PMID- 15795860 TI - Sexual conflict in the wild: elevated mating rate reduces female lifetime reproductive success. AB - Sexual conflict over mating rate is suggested to play a pivotal role in male female coevolution, and females are predicted to reject superfluous mating attempts. Recent work suggests that direct effects of multiple mating on female fitness are not fully understood. A major concern in studies of sexual conflict is how well the data obtained under controlled laboratory settings relate to natural conditions. We tested the effect of female multiple mating on reproductive success in a natural population of a polyandrous spider, Stegodyphus lineatus. Previous studies show that a male who succeeds in entering a female nest also mates with her; therefore, we used male encounter rate as a proxy of female mating rate. We further elevated female mating rate by introducing males into females' nests. Female lifetime reproductive success was assessed as the likelihood of successful reproduction, offspring production, and juvenile offspring body mass. Increased mating rate increased the time to oviposition and reduced the likelihood of successful reproduction. Female mating rate negatively affected offspring body mass. Manipulated females produced fewer offspring than control females. The observed patterns imply a net cost of polyandry to females and suggest that natural mating rates can be suboptimal for females under natural conditions. PMID- 15795859 TI - The evolution of infidelity in socially monogamous passerines: the strength of direct and indirect selection on extrapair copulation behavior in females. AB - Many studies have been aimed at understanding the maintenance of female infidelity in socially monogamous birds. Because engaging in extrapair copulations (EPCs) is believed to be costly for females, it has been argued that EPC behavior must bring indirect benefits to females by elevating offspring fitness. We use empirical data from the literature to assess the relative strength of indirect and direct selection on female EPC behavior, using quantitative genetic approximations of selection. This analysis confirmed that there is generally negative direct selection on EPC behavior caused by depressed paternal investment by social males. In contrast, there was no significant positive indirect selection on EPC behavior in females. A comparison between the two types of selection suggests that the force of direct negative selection is generally much stronger than that of indirect positive selection. Indirect selection is thus unlikely to maintain EPC behavior in the face of direct selection against it. We suggest that EPCs may instead be the result of antagonistic selection on loci influencing the outcome of male-female encounters and that EPC behavior per se may not be adaptive for females but may reflect sexual conflict due to strong selection in males to achieve extrapair copulations. PMID- 15795861 TI - Why are males bad for females? Models for the evolution of damaging male mating behavior. AB - One explanation for the cost to mating for females caused by damaging male mating behavior is that this causes the females to adaptively modify their subsequent life histories in a way that also increases male fitness. This might occur because the reduction in residual reproductive value of the female increases her optimal oviposition rate or because an increase in the current level of damage increases the female's optimal remating interval. In this article, I present models of a stochastic dynamic game in which males choose the level of mating damage that they inflict on females and females choose their oviposition rate and whether to remate. The models show that some level of damage is always an evolutionarily stable strategy and may even provoke females into making terminal reproductive investment (and hence a semelparous life history), that nondamaging populations are always invaded by damaging male mutants, and that damage evolves because of its effect on oviposition rate and despite its effect on remating interval. PMID- 15795862 TI - Escalation, retreat, and female indifference as alternative outcomes of sexually antagonistic coevolution. AB - Verbal and quantitative genetic models of sexually antagonistic coevolution suggest that coevolutionary arms races should be common. Sexual selection favors exaggeration of male persistence traits that are costly to females, and females, in turn, are selected to resist these traits. The heightened resistance by females is thought to then favor further exaggeration in the male trait, leading to an escalating coevolutionary arms race between persistence and resistance traits. Much of this theory, however, is based on an (implicit) assumption that there are tight constraints on how female resistance can evolve. We develop a theory that identifies and relaxes these constraints, allowing female resistance to evolve in a fashion that better reflects known empirical patterns in the evolution of female preference functions (the resistance trait). Our results suggest that evolutionary arms races that lead to the exaggeration of persistence and resistance will be much less common than formerly predicted. Females sometimes evolve indifference to male traits rather than resistance and can even evolve to discriminate against these traits. These alternative outcomes depend on the existence of genetic variance in the components of the female sensory system underlying female resistance and on the strength of natural selection acting on these components. Female indifference tends to evolve when natural selection on the sensory system is weak, and under these conditions, sexually antagonistic coevolution tends not to reduce female fitness significantly at equilibrium. When natural selection on the female sensory system is strong, however, then arms races are more likely, and female fitness is then sometimes significantly depressed at equilibrium. Sexually antagonistic coevolution is thus likely to have strong deleterious effects on population fitness only when female sensory traits are under strong natural selection to perform functions in addition to those involved with mating. Together, these results suggest that identifying the nature of genetic variation in and the strength of natural selection on female resistance should be a central goal of future studies of sexual conflict. PMID- 15795863 TI - Female mate preference and sexual conflict: females prefer males that have had fewer consorts. AB - Different aspects of male quality as a mate can vary independently. When this is the case, females may need to use multiple cues to accurately assess overall mate quality. We examined the pattern of mating preference in the cockroach Nauphoeta cinerea. Sexual conflict occurs in this species because male manipulation of female mating receptivity can result in reduced female fitness. We predicted that since females cannot remate within a single reproductive bout because of male manipulation, females should assess male mating history to avoid mating with males with low fertility caused by sperm exhaustion. In a mate-preference experiment, we found that females discriminated against males who had mated multiple times. Females also discriminated against males who had consorted with several females but had been prevented from mating with those females. Thus, females appeared to be able to detect cues on males that were derived from previous mates and to use this information to avoid mating with sperm-exhausted males. We suggest that females may commonly use multiple cues to assess different aspects of mate quality, especially when male quality changes over time. This is an under-studied phenomenon that could help explain individual variation in female mating preferences. PMID- 15795864 TI - Experimental removal and elevation of sexual selection: does sexual selection generate manipulative males and resistant females? AB - Sexual conflict over reproduction can occur between males and females. In several naturally promiscuous insect species, experimental evolution studies that have enforced monogamy found evidence for sexual conflict. Here, we subjected the naturally promiscuous, sperm-heteromorphic fruit fly Drosophila pseudoobscura to enforced monogamy, standard levels of promiscuity, and elevated opportunities for promiscuity in four replicate lines. We examined the effect of male and female selection history and the proximate effect of variation in male density on female fitness parameters. We found that male density rather than male selection history explained a greater degree of female fecundity, egg hatching success, and productivity. Additionally, females selected under elevated promiscuity had greater fecundity and hatching success than did enforced monogamy females. Selection line males do not differ in their capacity to coerce females to remate, suggesting no divergence in precopulatory manipulative ability. However, these males did vary in their ability to suppress female remating, suggesting postcopulatory manipulation. These results indicate that sexual conflict can be manifested through both the proximate effects of male density and the historical levels of sexual selection and that the sexes respond differentially to these factors and further stress the multifarious channels of sexual communication that contribute to fitness. PMID- 15795865 TI - Quantitative genetic models of sexual conflict based on interacting phenotypes. AB - Evolutionary conflict arises between reproductive partners when alternative reproductive opportunities are available. Sexual conflict can generate sexually antagonistic selection, which mediates sexual selection and intersexual coevolution. However, despite intense interest, the evolutionary implications of sexual conflict remain unresolved. We propose a novel theoretical approach to study the evolution of sexually antagonistic phenotypes based on quantitative genetics and the measure of social selection arising from male-female interactions. We consider the phenotype of one sex as both a genetically influenced evolving trait as well as the (evolving) social environment in which the phenotype of the opposite sex evolves. Several important points emerge from our analysis, including the relationship between direct selection on one sex and indirect effects through selection on the opposite sex. We suggest that the proposed approach may be a valuable tool to complement other theoretical approaches currently used to study sexual conflict. Most importantly, our approach highlights areas where additional empirical data can help clarify the role of sexual conflict in the evolutionary process. PMID- 15795866 TI - [Inborn errors of metabolism with neurological symptomathology in the neonatal period]. AB - INTRODUCTION: Congenital metabolic diseases are considered as rare diseases because of their low incidence and their clinical symptoms at onset. Sometimes they can just begin in the neonatal period. Their progressive knowledge and the availability of specific and sensitive biochemical procedures allow us to diagnose many congenital metabolic diseases, which were not recognized some years ago. PATIENTS AND METHODS: We reviewed the 52 patients with congenital metabolic diseases diagnosed for the last 25 years in our centre, evaluating the clinical presentation, neurological symptoms, complementary exams and clinical evolution. RESULTS: The mean age at onset of symptoms was 5 days and the mean age at diagnosis was 88 days of age. We considered a first group of 36 patients with inborn errors of intermediary metabolism, in whom hypotonia, weight loss and seizures are the main symptoms. The second group was composed of 8 patients with defective energy metabolism, who showed abnormal respiratory rhythm and hypotonia. Finally, we considered 8 patients with diseases of the complex molecules, who presented with hypotonia and cataracts as common symptoms at onset. The more common neurological symptoms in this period were hypotonia (60%), sensorial deficit (35%) and refractory seizures (23%). The complementary laboratory tests in the first phases of the diseases allowed us to suspect a congenital metabolic disease especially among intermediary and energy defects. EEG registration and CSF samples were important to diagnose some inborn errors of intermediary metabolism. In the first steps, the neuroimaging was less orientative, even if it allow the exclusion of other diseases. More than half of the patients with inborn errors of metabolism with onset in the neonatal period died within the first year of life. CONCLUSION: It is really important to suspect these diseases in the neonatal period so as to achieve an early diagnosis and therapy which may reduce the morbimortality. PMID- 15795867 TI - [Hospital care of cerebrovascular accident and the state of patients 12 months after]. AB - OBJECTIVES: Cerebrovascular accident should be of key importance due to its magnitude in terms of mortality and disability. In this study we describe hospital care of patients and follow them one year after. The aims is to uncover areas of improvement in the care of patients. PATIENTS AND METHODS: Observational study of a randomized sample of 535 patient with a diagnosis of cerebrovascular disease, during de acute phase and 12 months after, using clinical records and telephone interview. RESULTS: Thirty five percent of patients arrived within 6 hours of the occurrence of the event. Thirty six percent had a CAT/MNR within 6 hours. Mortality at hospital was 13.8% increasing up to 26% at 12 months. At discharge 49% had a neurological deficiency. At 12 months 35.8% of the survivors interviewed showed a Barthel Index of less than 95 points. CONCLUSIONS: Organizational measures that guarantee a quick and systematic assessment of brain lesions, early diagnosis and active therapeutic offer, have to be implemented. In the sample studied, only 3% of the patient were candidates to thrombolytic therapy. Rehabilitation can and should play a more relevant role in the prevention of sequelae. PMID- 15795868 TI - [Attention and executive function tests components in attention deficit/hyperactivity children]. AB - INTRODUCTION: It has been suggested that children with attention deficit/hyperactivity disorder (ADHD) have problems in attention and executive functions. A factor analysis showed that ADHD children had a different factor structure of the executive functions when they were compared with controls. AIM. To determine the factor structure of attention and executive functions in a group of ADHD children compared with a group of controls. SUBJECTS AND METHODS: 249 ADHD children and 372 controls of both genders, aged 6 to 11 years old, and from the different socio economic strata of Medellin city, were selected. Attention - Mental Control and Auditory Continuous Performance Test (ACPT)-- and executive functions --Phonologic and Semantic Verbal Fluency (FAS)--, and a shortened version of the Wisconsin Card Sorting Test (WCST) were assessed in both groups. Factor analyses were developed with principal components procedure were developed for each group and the total sample, using principal component procedure with orthogonal rotation. RESULTS: Stable factor structures were found for each group and for the total sample. The general factor structure was constituted by three independent factors: 1- Categorization, 2- Verbal fluency and verbal sustained attention, and 3- Cognitive flexibility. This factor structure explained 74.9% of the variance. CONCLUSION: The factor structures suggested that the components of attention and executive functions are similar for ADHD and control groups. The differences between groups would be quantitative. PMID- 15795869 TI - [Evaluation of cognitive impairment in a population of patients with Parkinson's disease by means of the mini mental Parkinson test]. AB - INTRODUCTION: Parkinson's disease (PD) is a neurodegenerative disorder in which motor symptoms are initially predominant; however, as the disease progresses, cognitive deterioration becomes more evident. Tests designed to detect cognitive impairment are therefore needed. AIMS: The aim of the study was to assess the cognitive status of a population of patients with PD by means of the Mini Mental Parkinson test (MMP). PATIENTS AND METHODS: In 110 patients with idiopathic PD, with a mean age of 66.4 +/- 10.2 years and an average history of 8.8 +/- 5.8 years of the disease, the mean severity of the disease as measured by the unified PD rating scale (UPDRS-III) was 26.2 +/- 12.9. RESULTS: There were 20 (18.2%) subjects with dementia according to DSM-IV criteria, with a score of 20.4 +/- 6.4 on the MMP. Of the 90 patients who did not have dementia, 47 cases displayed cognitive impairment, without the presence of disorders affecting their activities of daily living, who scored 25.3 +/- 4.7 points on the MMP, and 43 cases did not show any kind of cognitive impairment, with 28.0 +/- 3.3 points on the MMP; these differences were statistically significant p < 0.001. Results according to age brackets showed a decrease in MMP with age and more especially so concerning memory functioning. As to severity, there is a cognitive decline, in particular with regard to memory, orientation and abstraction. CONCLUSIONS: Cognitive capacity is seen to decline with age and the severity of the disease. PMID- 15795870 TI - [Clinical, electrophysiological and immunological evaluation of the response to treatment with intravenous immunoglobulins in several immune-mediated neuropathies]. AB - INTRODUCTION AND AIMS: Chronic immune-mediated neuropathies are characterised by their being predominantly demyelinating, by being associated to specific antibodies and by their response to immunotherapy. We evaluated the clinical, electrophysiological and immunological responses following treatment with intravenous immunoglobulins (IVIg) in different clinical forms. CASE REPORTS: We report on three patients with: 1. Multifocal motor neuropathy, 2. Multifocal sensory-motor neuropathy, and 3. Hypertrophic brachial plexopathy, who were evaluated before and 14 days after treatment with IVIg by means of clinical scales (MRC, Rankin), electrophysiological studies (ENG-EMG), and antibody (antiganglioside) determination. The three patients showed clinical improvement (> 20% MRC, > 1 Rankin) between the 4th and 7th day after the infusion, and this was maintained for 4-16 weeks. The ENG in cases 1 and 2 revealed conduction block (CB) in some nerves, lowered amplitude in others, and signs of demyelination. Following treatment, the CB disappeared in some nerves, while in others there was an improvement in the distal amplitude (distal block), but with no correlative improvement in the proximal amplitude, revealing new CBs that had previously gone undetected. The sum of post-treatment amplitudes improved in cases 1 and 2. In case 3, we were unable to demonstrate the existence of CBs, although we believe that they did exist proximally, and we found indirect signs of CB in EMG. The three cases had increased antiganglioside IgM antibodies levels, which did not undergo any significant variations. CONCLUSIONS: A 'consistent' clinical improvement was observed following treatment with IVIg. There was no correlation between the electrophysiological response (although apparent) and the degree of clinical response. CBs exist at different levels in the same nerve, which can be revealed after treatment. Following therapy, antiganglioside antibody levels remain high. PMID- 15795871 TI - [Alternating hemiplegia of childhood treated as epilepsy. Two new cases]. AB - INTRODUCTION: Alternating hemiplegia (AH) of childhood is a rare disease that gives rise to transient attacks of hemiplegia that may affect either side of the body indistinctly, or even both sides at the same time. Onset occurs before the age of 18 months and in some cases there are neonatal symptoms such as abnormal eye movements, especially nystagmus, and dystonic or tonic seizures. Attacks of hemiplegia, which disappear during sleep, begin before the age of one year. These symptoms can initially be taken for an epileptic disorder. CASE REPORTS: We describe the study of two male patients whose clinical symptoms appeared at the age of 6 months, with tonic seizures, upward deviation of gaze, without loss of consciousness, which occurred with a recurrence rate of between one and two attacks a week or several times a day. They were initially treated with antiepileptic drugs (AED), although the complementary tests, including electroencephalogram (EEG), computerised axial tomography (TAC), magnetic resonance (MR) scans and metabolic tests, were all normal. CONCLUSIONS: Since, to date, no specific test has been confirmed for the disease, diagnosis is essentially clinical and by exclusion. Treatment is symptomatic using flunarizine. In one of our cases, administration of a single 10 mg/day dose this drug at night lowered the frequency of the attacks of hemiplegia. The possibility of treating a patient who is a carrier of an AH with AED makes early knowledge and identification of this disease necessary in order to be able to improve the patient's prognosis. PMID- 15795872 TI - [Sequential changes in magnetic resonance in a limbic status epilepticus]. AB - INTRODUCTION: Transient alterations have been described in neuroimaging (MRI) studies of the non-convulsive focal status (NCFS). We report a case of NCFS together with the MRI findings. CASE REPORT: We describe the case of a 63-year old female who had a sister and two female cousins with epilepsy; the patient was admitted to hospital after being in state of confusion for 72 hours. Two similar bouts of delirium were reported as having occurred in the 2 preceding years, both of which lasted only a few minutes, and at that time a cardiology study, EEG and MRI scans of the head were performed with normal results. The EEG was compatible with left temporal status and MRI, and presented hyperintensity in the left temporal lobe in T2 and Flair, with no mass effect, with gadolinium uptake in leptomeninges and cortex. CSF was acellular and there were high protein levels in cerebrospinal fluid with a value of 1 g/dL. The patient's situation continued for 10 days, and did not respond initially to antiepileptic treatment. Temporal NCFS was diagnosed, with a cryptogenic rather than idiopathic aetiology. A preliminary MRI scan was normal and another scan performed 10 days after resolution showed a clear regression of the lesion. We related these findings to vasogenic and cytotoxic oedema secondary to the status. An MRI scan carried out at 3 months was normal. CONCLUSIONS: This case lends support to reports, in relation to the appearance of NCFS, of MR images compatible with oedema secondary to rupture of the blood-brain barrier. We base it on sequential MRI studies and on high protein levels in CSF. PMID- 15795873 TI - [Barkley's model of self-regulation applied to attention deficit hyperactivity disorder: a review]. AB - AIM: The aim of this study was to carry out a review of Barkley's model of self regulation and executive functions (EF) applied to attention deficit hyperactivity disorder (ADHD). DEVELOPMENT AND CONCLUSIONS: Our starting point was the discrepancy between the description and the explanation of the disorder; that is to say, on the one hand important advances have been made from the applied point of view, but on the other hand we still do not have an overall explanation of the problem. The causes of ADHD are then reviewed as regards both specific aetiological factors and the wider reaching explanatory models. In this latter case, as antecedents and contributions prior to the model of self regulation, we examine Douglas's attentional model and three models based on the concept of behavioural inhibition: Quay's neurobehavioural, Schachar's competitive, and Sergeant and van deer Meere's energetic models. Barkley's model was initially based on 'behavioural inhibition', but eventually the limitations that were detected and new lines of research led him to advance towards self regulation. The main components of the model are reviewed: behavioural inhibition processes, the very concept of self-regulation/self-control, the EF involved, and motor control. The model is applied to the behaviour and characteristics of children with ADHD, and its implications in the fields of evaluation and treatment are discussed. Finally, we outline some of its main controversies and, in the conclusions, its strong and weak points are highlighted. PMID- 15795874 TI - [Agrammatism and its symptoms]. AB - INTRODUCTION: We present a review the issue of agrammatism in order to examine its symptoms, the regularity with which they are observed, and the proposals suggested within the framework of Cognitive Neuropsychology to account for them. DEVELOPMENT: First, we focus on the debate regarding the status of agrammatism as an aphasic category with both theoretical and clinical validity, presenting two confronted views. On the one hand, the view that argues against the category of agrammatism due the variable performance of agrammatic patients in linguistic tasks. On the other hand, we present the view of those authors who defend, despite the variability, the notion of agrammatism as an aphasic syndrome. In the second section of the paper, we discussed the different symptoms that, on the basis of the available evidence, have been associated to agrammatism and the proposed explanations. We start by discussing the symptoms that agrammatic patients present in language production; in particular, we highlight the following: a) problems related to grammatical morphemes; b) a reduced sentence length; c) a noun-verb dissociation, with a better performance with nouns; d) difficulties in sentence construction, and finally, e) alteration in word order. Then, we discuss the symptoms that agrammatic patients have shown in language comprehension. CONCLUSION: In general, on the basis of the different agrammatic symptoms we have discussed, we favour the view of agrammatism as multicomponential syndrome rather than unitary one, with deficits in both morpholexical and syntactic language components. PMID- 15795875 TI - [Spontaneous dissection of the middle cerebral artery. A case report]. PMID- 15795876 TI - [Meningitis caused by Listeria monocytogenes in a patient with myasthenia gravis being treated with tacrolimus]. PMID- 15795877 TI - [Migraine that begins in old age]. PMID- 15795878 TI - Electrospray ionization mass spectrometry for identification and structural characterization of pregnane glycosides. AB - Pregnane glycosides are a class of naturally occurring substances characterized by some interesting biological activities and widely distributed in the plant kingdom and in some marine organisms. Their toxicity and use in herbal drugs and folk medicines has generated great interest in the chemical characterization of these molecules. In the study reported here the potential of electrospray ionization mass spectrometry (ESI-MS) in the identification and structural characterization of pregnane glycosides was examined. ESI-MS/MS and ESI-MS(n) analyses were performed on 27 different compounds employing two mass spectrometers equipped with a triple-quadrupole or an ion-trap analyzer. The data illustrate the ability of the ESI techniques in the identification of pregnane glycosides, including the nature of the pregnane core, the kind of ester substituents, the types of sugar residues (hexose, deoxyhexose, dideoxyhexose, O methyldeoxyhexose and O-methyldideoxyhexose), and the primary structure of the saccharide chain. From these data, a generalized fragmentation pathway was proposed by comparing the spectra acquired for all the compounds. Interestingly, similar results were obtained from the two instruments, thus demonstrating that detailed analyses of product ion spectra obtained using a triple-quadrupole mass spectrometer led to structural information comparable to those obtainable in MSn experiments using an ion trap. Different and complementary information was deduced by fragmenting the [M+H]+ or the [M+Na]+ ions, or the protonated aglycone [Agl+H]+ generated by in-source fragmentation. The present evidence clearly suggests that, in order to obtain a complete characterization of pregnane glycosides by MS, all three of these species should be accurately analyzed. PMID- 15795880 TI - High cumulative rate of secondary leukemia after continuous etoposide treatment for solid tumors in children and young adults. AB - BACKGROUND: In a national pediatric case-control study, we observed a very high relative risk of leukemia in patients who had received continuous etoposide (CE) over 6 months or more, but we could not estimate the absolute risk. The purpose of the present study was to estimate this absolute risk after CE. PROCEDURES: We report a study of 18 patients with refractory or recurrent tumors who received CE over 6 months or more between 1995 and 1997. It was administered either 3 days a week for 3/4 weeks ("3 x 3", 14 patients) or 7 days a week for 3/4 weeks ("7 x 3", four patients). RESULTS: Five patients developed secondary leukemia 10-25 months after the initiation of CE. All the others died of their first tumor. The cumulative incidence of leukemia at 30 months was 28% (95% CI, 10%-53%). A chromosome 11q23 rearrangement was found in 3/5 cases. All four patients who received the "7 x 3" CE schedule developed leukemia compared to 1/14 treated with the "3 x 3" CE schedule (P = 0.002). CONCLUSIONS: Given its efficacy, CE may still have a place as a palliative treatment. However, the risk of leukemia must be borne in mind when considering its use in patients with a better prognosis. PMID- 15795881 TI - Risk factors for and prevention of tobacco use. AB - This study describes the risk factors for and prevention of tobacco use among young people. Risk factors show differential importance along the course of tobacco use development. The contents of an effective prevention effort vary as a function of the age of target group. However, the mediation of the effects of effective programs is not well known. Various modalities of implementation of prevention programming may be important, though a majority of program evaluation has occurred in the school context. The majority of well-evaluated tobacco use prevention programs that employ a comprehensive social influences model show effects over an average of 6 years post-programming. PMID- 15795882 TI - Clinical and molecular characteristics of pediatric gastrointestinal stromal tumors (GISTs). AB - BACKGROUND: To describe the clinical characteristics, molecular features, treatment, and outcome of six pediatric patients with gastrointestinal stromal tumors (GISTs). PATIENTS AND METHODS: Retrospective clinical review of GISTs, seen at The Hospital for Sick Children (HSC) Toronto, over an 11-year period. All specimens were stained for the CD 117 and CD 34 antigens. Three specimens were sequenced for mutations in exons 9, 11, and 13 of the c-kit gene. RESULTS: Five patients were evaluated and treated at HSC and one was referred for histopathological consultation only. The median patient age at diagnosis was 13.6 years, (6.9-14.8 years); four were female. All patients presented with anemia secondary to gastrointestinal (GI) bleeding. The disease was localized in five patients and two had other malignancies consistent with the diagnoses of Carney's triad. Immunohistochemical staining for CD 117 and CD 34 showed heavy cytoplasmic localization in all of the tumor cells. A novel point mutation of KIT in codon 456 of exon 9 was found in one case. Complete surgical resection was achieved in the five patients managed at our center and none received adjuvant therapies. Disease recurred locally in one patient. Four patients are alive and one is lost to follow-up. CONCLUSIONS: In children and adolescents, GISTs should be considered in the differential diagnosis of anemia secondary to GI hemorrhage. The absence of an exon 11 mutation and the identification of a novel mutation in exon 9 suggest that pediatric GISTs may respond differently to currently available targeted therapies and therefore should be studied within the context of collaborative group trials. PMID- 15795883 TI - Medical utilization by immunodeficient, hemophiliac, and HIV positive children during summer camp: evidence for a safe camper experience. AB - BACKGROUND: Continuation of normal activities is vital to psychosocial development of children with serious illnesses. The purpose of this study was to determine whether or not it was safe for HIV-positive children and children with other immunodeficiencies to attend camp. PROCEDURE: The study population consisted of HIV (+) children, HIV negative siblings, and other immunodeficient campers attending Barretstown Gang (BG) Camp between 1998 and 2002. Their visit frequency to the on-site medical facility was compared within the study population and between 2,323 contemporaneous campers with cancer. RESULTS: Over half of the HIV (+) children were on active therapy. Greater than 97% of staff (49/51) made at least one visit compared with 64% (149/233) of campers (P < 0.04). HIV (-) siblings had almost the same need for medical attention (total visits) as children with immunodeficiencies (P = 0.34). Most visits [88%] among all diagnostic groups except hemophilia were non-disease related (328 vs. 47). Apart from URIs, there were few other infections and no fevers in the HIV(+) or immunodeficiency group, nor were there significant bleeds in the hemophiliacs. Most visits were for routine camp-type ailments. CONCLUSIONS: Our findings suggest that it is safe for HIV (+) and immunodeficient children to attend a properly staffed camp. PMID- 15795884 TI - Role of afterload reduction in the prevention of late anthracycline cardiomyopathy. AB - Afterload reduction is useful for treating and preventing adult congestive heart failure outside the realm of anthracycline toxicity. Afterload reducers are commonly used as treatment for acute heart failure associated with anthracyclines. However, the use of afterload reduction for the prevention of anthracycline associated late cardiomyopathy remains a controversial area, with strong opinions expressed concerning the potential risks and benefits of such an approach. This article will define "afterload" and the tools utilized to measure afterload. The study will then describe the theory for the use of afterload reduction in patients with anthracycline exposure who show signs of anthracycline cardiotoxicity, and will present data from the ACE-Inhibitor After Anthracyclines (AAA) Trial to help answer the question, "Should an afterload reducer be used to aid in preventing late anthracycline cardiotoxicity?" Finally, recommendations for future research will be presented. PMID- 15795885 TI - American Society of Pediatric Hematology/Oncology 18th Annual Meeting, Washington, D.C., May 13-16, 2005. Abstracts. PMID- 15795886 TI - Prevalence and etiology of acquired anemia in Medieval York, England. AB - This paper presents three distinct models for the development of acquired anemia: iron-deficiency anemia produced by the inadequate intake and/or absorption of iron, the anemia of chronic disease (ACD) caused by the body's natural iron withholding defense against microbial invaders, and megaloblastic anemia caused by insufficient intake and/or absorption of vitamin B(12) or folic acid. These etiological models are used to interpret the distribution and etiology of anemia among adult individuals interred at the Medieval Gilbertine Priory of St. Andrew, Fishergate, York (n = 147). This bioarchaeological analysis uncovered not only a strong relationship between decreasing status and increasing prevalence of anemia for both men and women, but also identified clear sex-based differences at this site. Within the high-status group, blood and iron loss as a result of rampant parasitism likely produced an environment ripe for the development of iron deficiency anemia, while the parasitic consumption of vitamin B(12) may have caused occasional cases of megaloblastic anemia. As status decreases, the interpretation of anemia becomes more complex, with megaloblastic anemia and ACD emerging as viable, potentially heavy contributors to the anemia experiences of low-status people at St. Andrew's. Apart from status effects, women (especially young women) are disproportionately affected by anemia when compared to men within their own status group and, on average, are also more likely to have experienced anemia than their male peers from other status groups. This suggests that high iron-demand reproductive functions helped to make iron-deficiency anemia a chronic condition in many women's lives irrespective of their status affiliation. PMID- 15795887 TI - Cross-cultural estimation of the human generation interval for use in genetics based population divergence studies. AB - The length of the human generation interval is a key parameter when using genetics to date population divergence events. However, no consensus exists regarding the generation interval length, and a wide variety of interval lengths have been used in recent studies. This makes comparison between studies difficult, and questions the accuracy of divergence date estimations. Recent genealogy-based research suggests that the male generation interval is substantially longer than the female interval, and that both are greater than the values commonly used in genetics studies. This study evaluates each of these hypotheses in a broader cross-cultural context, using data from both nation states and recent hunter-gatherer societies. Both hypotheses are supported by this study; therefore, revised estimates of male, female, and overall human generation interval lengths are proposed. The nearly universal, cross-cultural nature of the evidence justifies using these proposed estimates in Y-chromosomal, mitochondrial, and autosomal DNA-based population divergence studies. PMID- 15795888 TI - Effect of mangiferin on radiation-induced micronucleus formation in cultured human peripheral blood lymphocytes. AB - Irradiation causes a variety of lesions in important biomolecules of the cell through generation of free radicals leading to genomic instability. DNA strand breaks, acentric fragments, or defective kinetochores are manifested as micronuclei after the first cell division. Chemicals that can trap free radicals may reduce the deleterious effects of ionizing radiation. Mangiferin (MGN), a glucosylxanthone derived from Mangifera indica (mango), was investigated for its ability to reduce the frequency of radiation-induced micronucleated binucleate cells (MNBNCs) in cultured human peripheral blood lymphocytes (HPBLs). HPBL cultures were pretreated with 0, 5, 10, 20, 50, and 100 microg/ml of MGN for 30 min before exposure to 3 Gy of (60)Co gamma-radiation. The maximum decline in radiation-induced micronuclei was observed at a concentration of 50 microg/ml MGN; thereafter, a nonsignificant elevation in MNBNC frequency was observed at 100 microg/ml MGN. Since the lowest MNBNC frequency was observed for 50 microg/ml MGN, dose-response studies were undertaken using this concentration. Irradiation of HPBLs with 0, 1, 2, 3, or 4 Gy of gamma-radiation caused a dose-dependent elevation in the MNBNC frequency, while treatment of HPBLs with 50 microg/ml MGN 30 min before radiation resulted in significant declines in these frequencies. MGN alone did not alter the proliferation index. Irradiation caused a dose dependent decline in the proliferation index, while treatment of HPBLs with 50 micro/ml MGN significantly elevated the proliferation index in irradiated cells. MGN treatment reduced hydrogen peroxide-induced lipid peroxidation in HPBLs in a concentration-dependent fashion. In cell-free studies, MGN inhibited the induction of (.)OH (hydroxyl), O(2) (.-) (superoxide), DPPH (1,1-diphenyl-2 picrylhydrazyl), and ABTS(.+) (2,2-azino-bis-3-ethyl benzothiazoline-6-sulphonic acid) radicals in a dose-dependent manner. The results of this study indicate that MGN possesses radioprotective properties by suppressing the effects of free radicals. PMID- 15795889 TI - Prognosis of hepatocellular carcinoma: comparison of 7 staging systems in an American cohort. AB - Currently there is no consensus as to which staging system is best in predicting the survival of patients with hepatocellular carcinoma (HCC). The aims of this study were to identify independent predictors of survival and to compare 7 available prognostic staging systems in patients with HCC. A total of 239 consecutive patients with cirrhosis and HCC seen between January 1, 2000, and December 31, 2003, were included. Demographic, laboratory, and tumor characteristics and performance status were determined at diagnosis and before therapy. Predictors of survival were identified using the Kaplan-Meir test and the Cox model. Sixty-two percent of patients had hepatitis C, 56% had more than 1 tumor nodule, 24% had portal vein thrombosis, and 29% did not receive any cancer treatment. At the time of censorship, 153 (63%) patients had died. The 1- and 3 year survival of the entire cohort was 58% and 29%, respectively. The independent predictors of survival were performance status (P < .0001), MELD score greater than 10 (P = .001), portal vein thrombosis (P = .0001), and tumor diameter greater than 4 cm (P = .001). Treatment of HCC was related to overall survival. The Barcelona Clinic Liver Cancer (BCLC) staging system had the best independent predictive power for survival when compared with the other 6 prognostic systems. In conclusion, performance status, tumor extent, liver function, and treatment were independent predictors of survival mostly in patients with cirrhosis and HCC. The BCLC staging system includes aspects of all of these elements and provided the best prognostic stratification for our cohort of patients with HCC. PMID- 15795890 TI - Fluctuations in motor output during steady contractions are weakly related across contraction types and between hands. AB - The presence of differences in motor unit activity across contraction types and between hands suggests that the magnitude of fluctuations in motor output is only weakly related when these conditions are compared. Twenty right-handed young (24.1 +/- 5.3 years) and old (72.5 +/- 4.9 years) adults performed three levels ( 100 x 10(9)/L, a partial response (PR) as a rise in the platelet count > 50 x 10(9)/L, and a minor response (MR) as a rise in the platelet count < 50 x 10(9)/L. No response (NR) was defined as no increase in the platelet count. Because 4 patients were treated twice, a total of 39 outcomes of rituximab treatment were evaluated. Rituximab proved to be effective in 17 of 39 treatments [overall response 44% with 7 CR (18%) (1 patient showed a CR twice), 6 PR (15%), and 4 MR (10%)]. In 9/13 cases of CR or PR, the response (platelet level > 50 x 10(9)/L) was prompt, 1-2 weeks after the first infusion. The remaining patients responded 3-8 weeks later. Patients with CR and PR have been in remission for a median of 47 weeks. In general the side effects were few. In 2 cases, the treatment was stopped because of side effects either during or after the first infusion. Two fatal outcomes were recorded. A 71-year-old female with severe lung disease died 6 days after the first infusion of respiratory failure. The other patient, a 73-year-old man also with severe chronic obstructive lung disease, died of pneumonia approximately 13 weeks following the last rituximab treatment. It is concluded that rituximab may be a useful alternative therapy in patients with severe and symptomatic ITP refractory to conventional treatment. PMID- 15795921 TI - Hydroxychloroquine causes severe vacuolar myopathy in a patient with chronic graft-versus-host disease. AB - A 51-year-old man developed progressive debilitating limb and respiratory muscle weakness while undergoing treatment for chronic graft-versus-host disease secondary to allogeneic bone marrow transplant for mantle cell lymphoma. He had a normal serum creatine kinase level and acetylcholine receptor antibodies were negative. Electromyography showed a severe, nonirritable myopathy and a sensory motor axonal polyneuropathy. A muscle biopsy showed a necrotizing, vacuolar myopathy with many fibers containing autophagic and red-rimmed vacuoles, suggestive of an amphiphilic drug myopathy. The patient's strength and function improved significantly after discontinuation of hydroxychloroquine. PMID- 15795922 TI - Primary plasma cell leukemia: report of 17 new cases treated with autologous or allogeneic stem-cell transplantation and review of the literature. AB - Primary plasma cell leukemia (PPCL) is a rare hematologic malignancy characterized by the proliferation of plasma cells in blood, bone marrow, and other organs in the absence of established multiple myeloma. PPCL has a poor prognosis when treated with conventional therapy for multiple myeloma. We describe here 17 new cases of PPCL who underwent stem-cell transplantation (SCT) (2 cases observed by the authors and 15 cases from the International Bone Marrow Transplant Registry [IBMTR]). The first case was diagnosed in a 21-year-old male who presented with leukocytosis and acute renal failure. He was treated with hyper-CVAD, entered complete remission, and then proceeded to high-dose chemotherapy with peripheral stem-cell support. He is currently in complete remission 23 months after initial diagnosis and 19 months after autologous SCT. The second case was observed in a 31-year-old male who presented with leukocytosis and hepatic infiltration with plasma cells. He was treated with VAD chemotherapy and underwent allogeneic bone marrow transplantation from his HLA identical sister. He remained in complete remission for 3 years and then developed progressive refractory disease, dying 7 years after the initial diagnosis. In addition to these 2 cases, 15 further unpublished cases of PPCL from the IBMTR are reported here (treated between 1993 and 2001, 6 by autologous and 9 by allogeneic transplantation). Finally, the features of PPCL, the outcome, published data of SCT for PPCL, and indications for treatment are discussed. PMID- 15795923 TI - Efficacy of rituximab in an aggressive form of multicentric Castleman disease associated with immune phenomena. AB - Multicentric Castleman disease (MCD) is an uncommon lymphoproliferative disorder for which the best therapeutic option is not yet well established. Immune-related disorders are rare complications of MCD. We report on an MCD case in a 23-year old patient with extensive abdominal involvement and associated immune hemolytic anemia and Raynaud phenomenon. He was negative for human immunodeficiency virus (HIV) and human herpesvirus-8 (HHV-8). After 8 courses of the anti-CD20 monoclonal antibody (rituximab), the patient achieved complete remission. Interestingly, Raynaud phenomenon disappeared under treatment and no new hemolytic events occurred. Anti-CD20 antibody treatment could be an attractive therapeutic approach for MCD, mainly when immune-related disorders are associated. PMID- 15795924 TI - Temporal profiling of gene expression during neurogenesis and remodeling in the olfactory epithelium at short intervals after target ablation. AB - Neurogenesis in the olfactory epithelium (OE) is induced by olfactory bulbectomy (OBX), which effectively axotomizes olfactory sensory neurons (OSNs) and removes their synaptic targets, resulting in apoptosis. We used Affymetrix high-density oligonucleotide arrays to investigate changes in gene expression during initiation of signaling in pathways that regulate apoptosis and neurogenesis in the murine OE at 2, 8, 16, and 48 hr after bilateral OBX compared to that in sham operated controls. We focused on regulation of a defined set of genes associated with apoptosis, stem/progenitor cell regulation, and cell cycle progression because of the activation of these processes in OE degeneration and remodeling after OBX. After data scrubbing and categorical analysis, one-way analysis of variance identified 72 genes (4.9% of the present known genes) as being regulated significantly (P < 0.05) at one or more points; 50 were defined as regulated differentially with the false discovery rate at 10%. Significant changes in gene expression occurred in all categories as early as 2 hr post-OBX, with the greatest number of differentially regulated genes at 16 and 48 hr. Hierarchical cluster analysis and correlation coefficients were used to identify similarities in patterns of gene expression changes within and across categories. Validation was carried out with SuperArray macroarrays and real-time RT-PCR. Our results confirmed the participation of many genes in known signaling pathways and identified changes in the expression of 42 genes not identified previously as participating in apoptosis and neurogenesis in the OE. Additionally, our analyses indicated the early involvement of genes regulating cytoskeletal reorganization and angiogenesis in the response to OBX. These studies are an important first step in defining early time-dependent changes in gene expression after target ablation that lead to neurogenesis in the olfactory sensory epithelium. PMID- 15795925 TI - Molecular genetic confirmatory testing from newborn screening samples for the common African-American, Asian Indian, Southeast Asian, and Chinese beta thalassemia mutations. AB - beta-Thalassemia is a serious health problem in the United States, especially in California, due to increased Asian immigration. Neonatal screening by using high performance liquid chromatography (HPLC) or isoelectric focusing (IEF) may lead to confusion due to interactions of various hemoglobinopathies with beta thalassemia. Our purpose was to develop single-tube multiplexed PCR assays using original neonatal screening specimens to identify the mutations responsible for beta-thalassemia in order to expedite diagnostic confirmation. Primers were designed for two to six common ethnic-specific mutations using the amplification refractory mutation system (ARMS). This multiplex ARMS approach was standardized using DNA samples with known mutations for beta-thalassemia in those of Asian (Southeast Asian, Chinese, and Asian Indian) and African-American descent. Specimens from African-American neonates were tested for two mutations (-88 and 29); Asian Indians for five mutations (IVSI-1, IVSI-5, codons (Cd) 41/42, Cd 8/9, and 619-bp deletion); Chinese, Taiwanese, and Southeast Asians for seven mutations (Cd 41/42, Cd 17, -28, IVSII-654, Cd 71/72, IVSI-5, and IVSI-1). We identified each of these beta-thalassemia mutations in multiplexed ARMS from positive control samples. We tested 25 anonymized dried blood specimens from neonates who had been diagnosed with beta-thalassemia and who also belonged to these ethnic groups. We detected a mutation specific to the neonate's ethnic group using the ARMS approach in nearly all specimens, and the results were confirmed by sequencing. Multiplexed ARMS for ethnic-specific beta-thalassemia mutations from the original newborn screening dried blood specimens is a rapid and efficient approach for diagnostic confirmation. PMID- 15795926 TI - Design, synthesis, and application of azabicyclo[X.Y.0]alkanone amino acids as constrained dipeptide surrogates and peptide mimics. AB - Azabicyclo[X.Y.0]alkanone amino acids are challenging synthetic targets and useful tools for studying structure-activity relationships of native peptide ligands. They have been employed to increase potency and stability in conformationally rigid enzyme inhibitors and receptor ligands. Since last reviewed in 1997, activity in their synthesis and application has increased significantly and access is now available to a wider diversity of these peptide mimics. This review focuses on recent syntheses of these heterocyclic amino acids and their application in the investigation of biologically active peptides and peptide mimics. PMID- 15795927 TI - Underlying mechanisms of pronociceptive consequences of prolonged morphine exposure. AB - The opioid analgesics, commonly exemplified by morphine, represent the best option for the treatment of severe pain and for the management of chronic pain states, of both malignant and nonmalignant origin. It is well recognized that the prolonged use of opioids is associated with a requirement for ever-increasing doses in order to maintain pain relief at an acceptable and consistent level. This phenomenon is termed analgesic tolerance. While the concept that tolerance can develop as a result of cellular adaptations to the presence of the opioid has been proposed, it is now becoming abundantly clear that tolerance may also be related to a state of hyperalgesia that results from exposure to the opioid itself. Patients who receive long-term opioid therapy sometimes develop unexpected, abnormal pain. Similar paradoxical opioid-induced pain has been confirmed in a number of animal studies, even during the period of continuous opioid delivery. A number of recent studies have demonstrated that such pain may be secondary to neuroplastic changes that occur in the brain and spinal cord. One such change may be the activation of descending pain facilitation mechanisms arising from the rostral ventromedial medulla (RVM) elicited in part by increased activity of cholecystokinin (CCK) in the RVM. A cascade of pronociceptive events may follow, such as opioid-induced upregulation of spinal dynorphin levels that promotes enhanced input from primary afferent nociceptors. This mechanism appears to depend on intact descending pathways from the RVM, since interrupting this pathway abolishes enhanced abnormal pain. Furthermore, extended opioid exposure also can elicit increased calcitonin gene related peptide (CGRP) and substance P expression in the dorsal root ganglia. It is probable that increased pain elicited by opioids is a critical factor in the behavioral manifestation of opioid tolerance because the same manipulations that block abnormal pain also block antinociceptive tolerance. Taken together, such studies show that opioids elicit systems-level adaptations resulting in pain due to descending facilitation, upregulation of spinal dynorphin, and enhanced, evoked release of excitatory transmitters from primary afferents. These adaptive changes in response to sustained exposure to opioids indicate the need for the evaluation of the clinical consequences of long-term opioid administration. Additionally, these findings suggest a need for novel chemistry involving design of agents that may counteract opiate-induced neuroplastic adaptations resulting in pain relief without analgesic tolerance. PMID- 15795929 TI - Tau is not normally degraded by the proteasome. AB - Tau-positive inclusions in neurons are consistent neuropathologic features of the most common causes of dementias such Alzheimer's disease and frontotemporal dementia. Ubiquitinated tau-positive inclusions have been reported in brains of Alzheimer's disease patients, but involvement of the ubiquitin-dependent proteasomal system in tau degradation remains controversial. Before considering the tau degradation in pathologic conditions, it is important to determine whether or not endogenous tau is normally degraded by the proteasome pathway. We therefore investigated this question using two complementary approaches in vitro and in vivo. Firstly, SH-SY5Y human neuroblastoma cells were treated with different proteasome inhibitors, MG132, lactacystin, and epoxomicin. Under these conditions, neither total nor phosphorylated endogenous tau protein levels were increased. Instead, an unexpected decrease of tau protein was observed. Secondly, we took advantage of a temperature-sensitive mutant allele of the 20S proteasome in Drosophila. Genetic inactivation of the proteasome also resulted in a decrease of tau levels in Drosophila. These results obtained in vitro and in vivo demonstrate that endogenous tau is not normally degraded by the proteasome. PMID- 15795928 TI - Prospective characterization of neural stem cells by flow cytometry analysis using a combination of surface markers. AB - Neural stem cells (NSCs) with self-renewal and multilineage differentiation properties can potentially repair degenerating or damaged neural tissue. Here, we have enriched NSCs from neurospheres, which make up a heterogeneous population, by fluorescence-activated cell sorting (FACS) with antibodies against syndecan-1, Notch-1, and integrin-beta1, which were chosen as candidates for hematopoietic cell-or somatic stem cell-markers. Antigen-positive cells readily initiated neurosphere formation, but cells lacking these markers did so less readily. Doubly positive cells expressing both syndecan-1 and Notch-1 underwent neurosphere formation more efficiently than did singly positive cells. The progeny of sorted cells could differentiate into neurons and glial cells both in vitro and in vivo. These antibodies were also useful for isolating cells from the murine embryonic day 14.5 brain that efficiently formed neurospheres. In contrast, there was no distinct difference in neurosphere formation efficiency between Hoechst 33342-stained side population cells and main population cells, although the former are known to have a stem cell phenotype in various tissues. These results indicate the usefulness of syndecan-1, Notch-1, and integrin-beta1 as NSC markers. PMID- 15795930 TI - Regulation of NT-3 and BDNF levels in guinea pig auditory brain stem nuclei after unilateral cochlear ablation. AB - Injury to areas of the central nervous system can alter neurotrophin levels, which may influence postlesion neuronal survival and plasticity. To determine if sensorineural hearing loss induces such changes, we used an enzyme-linked immunosorbent assay (ELISA) to measure neurotrophin-3 (NT-3) and brain-derived neurotrophic factor (BDNF) levels in adult guinea pig brain stem auditory nuclei 3-60 days after a unilateral cochlear ablation (UCA). After UCA, which destroyed the cochlea and cochlear nerve on one side, NT-3 levels were usually depressed at 3 days by 22-44% but became elevated transiently at 7 days by 28-124%. BDNF levels were elevated transiently by 50% on the ablated side in the anteroventral (AVCN) and posteroventral (PVCN) cochlear nucleus at 3 days and may have signaled support for the survival of deafferented neurons. Coincident elevation at 3 and 7 days of BDNF or NT-3 and phosphorylated extracellular signal-regulated protein kinase 2 (ERK2-P) suggested a relationship to stimulated signal transduction activity. Elevated neurotrophin levels may have contributed to synaptogenesis in the AVCN and the superior olive and to changes in the synaptic biochemistry in the auditory nuclei after UCA. In contrast, deficiencies or failure to elevate neurotrophin levels within several days of the UCA correlated with upregulation of phosphorylated stress-activated protein kinase (SAPK-P), suggesting a relationship with stress-activated signal transduction and with the sparse degeneration of fibers observed in some of the auditory nuclei after UCA. PMID- 15795931 TI - Glucose-6-phosphate dehydrogenase and NADPH-consuming enzymes in the rat olfactory bulb. AB - The resistance to oxidative stress is a multifactorial reaction involving the clustering of transcriptionally regulated genes. Because glucose-6-phosphate dehydrogenase (G6PD), the principal enzyme responsible for reducing power, is highly expressed in the olfactory bulb (OB), it is of interest to verify whether other enzymes utilizing NADPH are also highly expressed. The level and localization of G6PD- and NADPH-consuming enzymes, such as NADPH-cytochrome P450 oxidoreductase (P450R), glutathione reductase (GR), and NADPH-diaphorase (NADPH d), were analyzed in the rat olfactory bulb (OB) by quantitative histochemistry and immunohistochemistry. The highest concentration of G6PD, P450R, and GR was observed in the olfactory nerve layer (ONL), suggesting a correlation in the expression of these enzymes at the gene level. Correlation in staining intensity between G6PD and NADPH-d activities occurred only in part of the ONL, some glomeruli, and scattered periglomerular cells. This peculiar distribution of NADPH-d could reflect a spatial patterning of the nose to bulb projections. Taken together, these results indicate that G6PD expression in the ONL could be related to the importance of generating a substantial supply of NADPH to sustain the detoxifying systems represented by GR and P450R reactions and, only in discrete zones, by NADPH-d activity. PMID- 15795932 TI - Drosophila melanogaster as a complementary system for studying HIV-1-related genes and proteins. AB - Human immunodeficiency virus type 1 (HIV-1) persists as a pandemic even though new information about the virus is being discovered on a daily basis. If the brain becomes infected, HIV-1 encephalitis or HIV-1-associated dementia may develop. There is much to be learned about the modes of action and mechanisms of genes and proteins, and their interactions that underlie HIV-1 infection. Drosophila melanogaster has been used successfully to study genes and proteins related to HIV-1 infection, including but not limited to the disturbance of antimicrobial responses by viral protein U and the identification of D. melanogaster analogs to the serine palmitoyltransferase 5 and 6 proteins that play a role in activation of transcription by the HIV-1 Tat protein in human cells. We believe that utilizing D. melanogaster as a complementary system for the study of genes and proteins related to HIV-1 infection will provide useful information that will lead to new studies designed to enhance our understanding of the mechanistic roles of these molecules. In the present study, we focus on the utilization of D. melanogaster as a complementary system for studying HIV-1 related genes and proteins, why this research should be extended, and why this complementary system is an important method for enhancing our understanding of the genetics involved in HIV-1 infection. PMID- 15795933 TI - Reduced CSF PLTP activity in Alzheimer's disease and other neurologic diseases; PLTP induces ApoE secretion in primary human astrocytes in vitro. AB - Phospholipid transfer protein (PLTP) plays a pivotal role in cellular lipid efflux and modulation of lipoprotein metabolism. PLTP is distributed widely in the central nervous system (CNS), is synthesized by glia and neurons, and is active in cerebrospinal fluid (CSF). The aims of this study were to test the hypothesis that patients with Alzheimer's disease (AD) have altered PLTP-mediated phospholipid transfer activity in CSF, and to examine the potential relationship between PLTP activity and apolipoprotein E (apoE) levels in CSF. We assessed PLTP activity and apoE concentration in CSF of patients with probable AD (n = 50), multiple sclerosis (MS; n = 9), other neurologic diseases (n = 21), and neurologically healthy controls (n = 40). PLTP activity in AD was reduced compared to that in controls (P < 0.001), with approximately half of the AD patients with PLTP activity values below all controls. Patients with MS had lower PLTP activity than AD patients (P < 0.001). PLTP activity was highly correlated with PLTP mass, as estimated by Western blot (r = 0.006; P < 0.01). CSF PLTP activity positively correlated with apoE concentration in AD (R = 0.435; P = 0.002) and controls (R = 0.456; P = 0.003). Anti-apoE immunoaffinity chromatography and Western blot analyses indicated that some CSF PLTP is associated with apoE-containing lipoproteins. Exogenous addition of recombinant PLTP to primary human astrocytes significantly increased apoE secretion to the conditioned medium. The findings of reduced PLTP activity in AD CSF, and the observation that PLTP can influence apoE secretion in astrocytes suggest a potential link between alterations in the brain lipid metabolism and AD pathogenesis. PMID- 15795934 TI - Systematic administration of iptakalim, an ATP-sensitive potassium channel opener, prevents rotenone-induced motor and neurochemical alterations in rats. AB - Our previous studies revealed that iptakalim, a novel ATP-sensitive potassium channel opener, has a significant neuroprotective function against ischemia in vivo or rotenone-induced neurotoxicity in vitro. To investigate the potential pharmaceutical benefit of ATP-sensitive potassium channel openers on neurodegenerative diseases, we studied the effects of iptakalim and diazoxide, a selective mitochondrial ATP-sensitive potassium channel opener, on the rotenone induced nigrostriatal degeneration in rats. Iptakalim (1.5 mg/kg/day, orally) or diazoxide (1.5 mg/kg/day, orally) alone was administered to rats for 3 days, and then for 4 weeks was used daily with an injection of rotenone (2.5 mg/kg/day, subcutaneously) 1 hr later each time. The results showed that rotenone-infused rats exhibited parkinsonian symptoms and had dopamine depletion in the striatum and substantia nigra. Pretreatment with iptakalim or diazoxide prevented rotenone induced catalepsy and the reduction of striatum dopamine contents. Moreover, iptakalim and diazoxide reduced the enzymatic activities and mRNA levels of inducible nitric oxide synthase elicited by chronic administration of rotenone. These neuroprotective effects of iptakalim and diazoxide were abolished by 5 hydroxydecanoate, a selective mitochondrial ATP-sensitive potassium channel blocker. In conclusion, our data suggested that mitochondrial ATP-sensitive potassium channels might play a key role in preventing both parkinsonian symptoms and neurochemistry alterations induced by rotenone in rats. The selective activation of mitochondrial ATP-sensitive potassium channels may provide a new therapeutic strategy for prevention and treatment of neurodegenerative disorders such as Parkinson's disease. PMID- 15795935 TI - Selective caspase activation may contribute to neurological dysfunction after experimental spinal cord trauma. AB - Caspases are implicated in apoptotic cell death after spinal cord injury (SCI), but the relative contribution of these proteases to the secondary injury process has been only partially described. We examined the activation of caspases 1, 2, 3, 6, 8, and 9 from 1 hr to 7 days after moderate contusion injury induced by a weight-drop method in the rat. Tissue homogenates from a 1-cm segment of cord that contained the site of impact were processed by fluorometric enzymatic activity assays and/or immunoblotting methods. Caspases 3, 8, and 9 were activated from 1 to 72 hr after injury, whereas caspases 1, 2, and 6 were not. Double-label immunohistochemistry utilizing antibodies for CNS cell-type-specific markers and active subunits of caspases 3, 8, or 9 showed that, at 4 and 72 hr after injury, these caspases were primarily activated in neurons and oligodendrocytes, rather than in astrocytes. Active caspase subunits were present in neurons within the necrotic lesion core at 4 hr after injury and in cells more than several segments away at 4 or 72 hr after injury. Intrathecal injection of the pan-caspase inhibitor Boc-Asp (OMe)-fluoromethylketone (Boc-d-fmk) at 15 min after injury improved locomotor function 21 and 28 days later. Treatment with the selective caspase 3 inhibitor N-benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethyl ketone (z-DEVD-fmk) improved function at 21 days after injury. These data suggest that caspases 3, 8, and 9 may be differentially activated in white and gray matter after spinal cord trauma and that such activation may contribute to subsequent neurological dysfunction. PMID- 15795936 TI - Inhibition of Cdk6 expression through p38 MAP kinase is involved in differentiation of mouse prechondrocyte ATDC5. AB - Because a temporal arrest in the G1-phase of the cell cycle is a prerequisite for cell differentiation, this study investigated the involvement of cell cycle factors in the differentiation of cultured mouse prechondrocyte cell line ATDC5. Among the G1 cell cycle factors examined, both protein and mRNA levels of cyclin dependent kinase (Cdk6) were downregulated during the culture in a differentiation medium. The protein degradation of Cdk6 was not involved in this downregulation because proteasome inhibitors did not reverse the protein level. When inhibitors of p38 MAPK, ERK-1/2, and PI3K/Akt were added to the culture, only a p38 MAPK inhibitor SB203580 blocked the decrease in the Cdk6 protein level by the differentiation medium, indicating that the Cdk6 inhibition was mediated by p38 MAPK pathway. In fact, p38 MAPK was confirmed to be phosphorylated during differentiation of ATDC5 cells. Enforced expression of Cdk6 in ATDC5 cells blocked the chondrocyte differentiation and inhibited Sox5 and Sox6 expressions. However, the Cdk6 overexpression did not affect the proliferation or the cell cycle progression, suggesting that the inhibitory effect of Cdk6 on the differentiation was exerted by a mechanism largely independent of its cell cycle regulation. These results indicate that Cdk6 may be a regulator of chondrocyte differentiation and that its p38-mediated downregulation is involved in the efficient differentiation. PMID- 15795937 TI - Enhanced angiogenesis characteristic of SPARC-null mice disappears with age. AB - The impairment of angiogenesis in aging has been attributed, in part, to alterations in proteins associated with the extracellular matrix (ECM). SPARC (secreted protein acidic and rich in cysteine/osteonectin/BM-40) is a matricellular protein that regulates endothelial cell function as well as cell ECM interactions. We have previously shown that angiogenesis, as reflected by fibrovascular invasion into subcutaneously implanted polyvinyl alcohol (PVA) sponges, is increased in SPARC-null mice (6-9 months of age) relative to their wild-type (WT) counterparts. In this study, we define the influence of aging on (a) the expression of SPARC and (b) fibrovascular invasion into sponge implants in SPARC-null and WT mice. The expression of SPARC in fibroblasts and endothelial cells derived from young donors (humans mean age less than 30 years and mice 4-6 months of age) and old donors (humans mean age over 65 years and mice 22-27 months of age) decreased 1.6 to 2.3-fold with age. Analysis of fibrovascular invasion into sponges implanted into old (22-27 months) SPARC-null and WT mice showed no differences in percent area of invasion or collagenous ECM. Moreover, sponges from old SPARC-null and WT mice contained similar levels of VEGF that were significantly lower than those from young (4-6 months) mice. In contrast to fibroblasts from young SPARC-null mice, dermal fibroblasts from old SPARC-null mice did not migrate farther, proliferate faster, or produce greater amounts of VEGF relative to their old WT counterparts. However, when stimulated with TGF beta1, primary cells isolated from the sponge implants, and dermal fibroblasts from both old SPARC-null and WT mice, showed marked increases in VEGF secretion. These data indicate that aging results in a loss of enhanced angiogenesis in SPARC-null mice, as a result of the detrimental impact of age on cellular functions, collagen deposition, and VEGF synthesis. However, the influence of aging on these processes may be reversed, in part, by growth factor stimulation. PMID- 15795938 TI - Caudal fin in the white shark, Carcharodon carcharias (Lamnidae): a dynamic propeller for fast, efficient swimming. AB - The caudal peduncle and caudal fin of Carcharodon carcharias together form a dynamic locomotory structure. The caudal peduncle is a highly modified, dorsoventrally compressed and rigid structure that facilitates the oscillations of the caudal fin. Its stiffness appears to be principally achieved by a thick layer of adipose tissue ranging from 28-37% of its cross-sectional area, reinforced by cross-woven collagen fibers. Numerous overlying layers of collagen fibers of the stratum compactum, oriented in steep left- and right-handed helices (approximately 65 degrees to the shark's long axis), prevent bowstringing of the perimysial fibers, which lie just below the dermal layer. Perimysial fibers, muscles, and the notochord are restricted to the dorsal lobe of the caudal fin and comprise the bulk of its mass. Adipose tissue reinforces the leading edge of the dorsal lobe of the caudal fin and contributes to maintaining the ideal cross sectional geometry required of an advanced hydrofoil. Most of the mass of the ventral lobe consists of the ceratotrichia or fin rays separated by thin partitions of connective tissue. Dermal fibers of the stratum compactum of the dorsal lobe occur in numerous distinct layers. The layers are more complex than in other sharks and appear to reflect a hierarchical development in C. carcharias. The fiber layer comprises a number of thick fiber bundles along the height of the layer and the layers get thicker deeper into the stratum compactum. Each of these layers alternates with a layer a single fiber-bundle deep, a formation thought to give stability to the stratum compactum and to enable freer movements of the fiber system. In tangential sections of the stratum compactum the fiber bundles in the dorsal lobe can be seen oriented with respect to the long axis of the shark at approximately 55-60 degrees in left- and right-handed helices. Because of the backward sweep of the dorsal lobe (approximately 55 degrees to the shark's long axis) the right-handed fibers also parallel the lobe's long axis. In the dorsal lobe, ceratotrichia are present only along the leading edge (embedded within connective tissue), apparently as reinforcement. Stratum compactum fiber bundles of the ventral lobe, viewed in transverse section, lack the well-ordered distinctive layers of the dorsal lobe, but rather occur as irregularly arranged masses of tightly compacted fiber bundles of various sizes. In tangential sections the fiber bundles are oriented at angles of approximately 60 degrees, generally in one direction, i.e., lacking the left- and right-handed helical pattern. Tensile load tests on the caudal fin indicate high passive resistance to bending by the skin. The shear modulus G showed that the skin's contribution to stiffness (average values from three specimens at radians 0.52 and 1.05) is 33.5% for the dorsal lobe and 41.8% for the ventral. The load tests also indicate greater bending stiffness of the ventral lobe compared to the dorsal. Overall, the anatomy and mechanics of the dorsal lobe of C. carcharias facilitate greater control of movement compared to the ventral lobe. The helical fiber architecture near the surface of the caudal fin is analogous to strengthening of a thin cylinder in engineering. High fiber angles along the span of the dorsal lobe are considered ideal for resisting the bending stresses that the lobe is subjected to during the locomotory beat cycle. They are also ideal for storing strain energy during bending of the lobe and consequently may be of value in facilitating the recovery stroke. The complex fiber architecture of the caudal fin and caudal peduncle of C. carcharias provides considerable potential for an elastic mechanism in the animal's swimming motions and consequently for energy conservation. PMID- 15795940 TI - HIV and youth. Spring break ushers in overload for HIV/AIDS program requests. PMID- 15795939 TI - Docosahexaenoic acid induces apoptosis in proliferating human endothelial cells. AB - n-3 polyunsaturated fatty acids (PUFAs) have been shown to exert beneficial effects in the prevention of cardiovascular disease, inflammation, and on tumor growth. To investigate effects of PUFAs on proliferation and apoptosis in endothelial cells, we tested the n-3 PUFA docosahexaenoic acid (DHA) and the n-6 PUFA arachidonic acid (AA) in human umbilical vein endothelial cells (HUVEC). The mitochondrial membrane potential (MMP) and the production of reactive oxygen species were examined by flow cytometry. Phosphorylation of p53 or p38 MAP kinase, and total levels of p53 were measured by Western blot. DNA binding activity of p53 was analyzed with a TransAM transcription factor assay kit. Tube formation was assessed on Matrigel. In proliferating HUVEC, but not in confluent cells, DHA reduced cell viability and induced apoptosis, as demonstrated by increases in membrane leakage (propidium iodide (PI) staining), Annexin-V binding, sub G(1) phase in the cell cycle, and TUNEL-positive cells. AA had no effect on these parameters. In addition to a reduced MMP and increased reactive oxygen species, phosphorylation of p38 and p53 (serine 15) and impaired DNA binding of p53 were observed. There was no change in total levels of p53. The p38 inhibitor SB203580 had no effect on Annexin V binding. DHA also attenuated HUVEC tube formation. Taken together, DHA induces apoptosis in proliferating, but not in resting HUVEC, potentially via the phosphorylation of p53, resulting in decreased p53 DNA binding. The results suggest that anti-angiogenic effects of DHA may be due to induction of apoptosis in proliferating endothelial cells. PMID- 15795941 TI - Medical marijuana. Medical necessity defense fails for man with AIDS. PMID- 15795943 TI - Survey shows intervention crucial to halt HIV in youth. PMID- 15795942 TI - HIV and youth. Youth HIV/AIDS cases. PMID- 15795944 TI - Vaccine. Scientists use antispam to assist AIDS vaccine work. PMID- 15795945 TI - Appeals court: doctor's fear of HIV-transmission unfounded. PMID- 15795946 TI - Will dispute. Court clarifies intent of AIDS patient's will. PMID- 15795947 TI - Criminal transmission. HIV-positive man charged with murder for transmitting virus. PMID- 15795948 TI - Serum albumin: a late-reacting negative acute-phase protein in clinically evident inflammation in dialysis patients. PMID- 15795949 TI - Drug interaction between sevelamer and cyclosporin. PMID- 15795950 TI - The development of the Community-based Nurse-midwifery Education Program: An innovation in distance learning. AB - Since the inception of nurse-midwifery education 80 years ago, educators have looked for ways in which to educate enough nurse-midwives. The application of distance learning to nurse-midwifery education expanded opportunities for a greater number of students to have access to an education in nurse midwifery. The story of the Community-based Nurse-midwifery Education Program is presented as an exemplar. PMID- 15795951 TI - Watchdog for the main artery. A new wireless device monitors dangerous aortic aneurysms. PMID- 15795952 TI - A cardiac "cap" provides rest for failing hearts. PMID- 15795953 TI - Bonuses from diabetes drugs. They may protect a much larger group of patients at risk. PMID- 15795954 TI - Complementary care. How statin alternatives stack up. Some help control cholesterol; others are potentially harmful. PMID- 15795955 TI - Soy appears to lose some luster. Heart-protective, or has-been? Here's a guide to new studies. PMID- 15795956 TI - Ask the doctors. In an emergency room, one doctor said that a highly sensitive blood test indicated that my chest pain was the result of a heart attack. A second doctor disagreed. He said that another standard type of blood test didn't show heart muscle damage, although the more sensitive test detected an abnormality. Which doctor was right? PMID- 15795957 TI - Sacroiliac joint dysfunction. A source of low back pain. PMID- 15795958 TI - Health tips. Keeping your fingernails healthy. PMID- 15795959 TI - Interest in new shoulder replacement technique growing. PMID- 15795961 TI - Is it better to stretch before I exercise or afterward? PMID- 15795960 TI - Raising HDL. In search of more 'good' cholesterol. PMID- 15795962 TI - Standards for Hospital Libraries 2002 with 2004 revisions. AB - The Medical Library Association's Standards for Hospital Libraries 2002 has been developed as a guide for hospital administrators, librarians, and accrediting bodies to ensure that hospitals have the resources and services to effectively meet their needs for knowledge-based information. Specific requirements for knowledge-based information include that the library be a separate department with its own budget. Knowledge-based information in the library should be directed by a qualified librarian who functions as a department head and is a member of the Academy of Health Information Professionals. The standards define the role of the medical librarian and the links between knowledge-based information and other functions such as patient care, patient education, performance improvement, and education. In addition, the standards address the development and implementation of the knowledge-based information needs assessment and plan, the promotion and publicity of the KBI services, and the physical space and staffing requirements. The role, qualifications, and functions of a hospital library consultant are outlined. The health sciences library is positioned to play a key role in the hospital. The increasing use of the Internet and new information technologies by medical, nursing, and allied health staffs, patients and the community require new strategies, strategic planning, allocation of adequate resources, and selection and evaluation of appropriate information resources and technologies. The Hospital Library Standards Committee has developed this document as a guideline to be used in facing these challenges together. PMID- 15795963 TI - [Evaluation of bloody nipple discharge]. AB - A 64-year old woman was referred for radiological evaluation of right sided bloody nipple discharge lasting for five months. Finally galactography supposed the diagnosis of an intraductal papilloma which was confirmed by surgery. Histopathologically an eight millimeter measuring intraductal papilloma with atypical ductal hyperplasia with signs of a carcinoma in situ was seen. PMID- 15795964 TI - A preliminary report on a skills-based telephone-administered peer support programme for patients with multiple sclerosis. AB - OBJECTIVE: Peer-support interventions have shown no statistically significant or clinically meaningful effect on quality of life (QOL) or depressive symptoms for multiple sclerosis (MS) patients. Peer-support interventions for MS generally provide support but no skills training. The aim of this study was to evaluate a brief telephone-administered skills-training model of peer-support for patients with MS. METHODS: Sixteen patients with MS showing signs of moderate distress received eight sessions of telephone-administered peer support (TAPS). TAPS is a manualized programme administered by peer-support counsellors diagnosed with MS. Using a workbook, peer-support counsellors teach skills to manage distress and MS symptoms. Subjective depression was assessed using the Center for Epidemiological Studies Depression Scale while objective depression was rated using the Hamilton Rating Scale for Depression. QOL was measured pre- and post-treatment using the SF-36. RESULTS: The participants showed significant improvements on both the CESD (p = 0.04) and the HRSD (p = 0.01). Overall QOL improved significantly (p = 0.045), however this was not reflected in either the Physical Health composite score or the Mental Health Composite Scale (p > 0.17). CONCLUSIONS: These findings suggest that TAPS may prove to be an efficacious peer-support model for patients with MS. PMID- 15795965 TI - Computational toxicology: framework, partnerships, and program development. September 29-30, 2003, Research Triangle Park, North Carolina. PMID- 15795966 TI - Spinal epidural lipomatosis: case reports, literature review and meta-analysis. AB - BACKGROUND CONTEXT: Symptomatic spinal epidural lipomatosis (SEL), a rare cause of spinal cord compression, has most often been associated with exogenous steroid use. PURPOSE: Identify four associations with SEL, correlate the associated groups with level of disease and compare treatment with outcome data in these groups. STUDY DESIGN/SETTING: Case reports of three patients and analysis of 104 cases from the literature. PATIENT SAMPLE: Three patients from the senior author's practice. OUTCOME MEASURES: Not applicable. METHODS: The authors report three new cases of SEL not associated with steroid use. They review all available English literature and present a table of all 104 reported cases. RESULTS: The clinical course of three new patients is reported. CONCLUSIONS: Associated conditions are exogenous steroid use, obesity, endogenous steroid excess, and some remain idiopathic. Although SEL is a rare condition, our review of the literature reveals many more reported cases than previously thought. With increased awareness of this condition and improved imaging techniques, further studies of this disease should be undertaken. PMID- 15795967 TI - Scoliosis in Steinert syndrome: a case report. AB - BACKGROUND CONTEXT: Steinert syndrome is described as an autosomal dominant condition characterized by progressive muscular wasting, myotonia, musculoskeletal manifestations and rare spinal defects. Little is reported about spinal deformity associated with this syndrome. PURPOSE: We present a patient with Steinert syndrome complicated by scoliosis. In the literature on muscular dystrophy, other than Duchenne, little mention is given to the problem of scoliosis in general and its treatment in particular. STUDY DESIGN: A case report of a patient with Steinert syndrome associated with thoracic scoliosis and hypokyphosis is presented. METHODS: A 17-year-old boy presented with King type II right thoracic scoliosis (T5-T11, Cobb angle of 40 degrees) and hypokyphosis--10 degrees. He was treated with posterior stabilization and instrumentation at level T3-L2 with a postoperative correction of the scoliotic curve to 20 degrees. Histopathologic examination of the muscles confirmed the diagnosis of Steinert myotonic dystrophy. RESULTS: At 30-month follow-up, the patient was clinically pain free and well balanced. Plain radiographs showed solid spine fusion with no loss of deformity correction. CONCLUSIONS: Scoliosis in Steinert syndrome shares the characteristic of an arthrogrypotic neuromuscular curve and demands the extensive soft tissue release for optimal surgical correction. Intraoperative observations included profound tissue bleeding, abnormally tough soft tissues and a difficult recovery from anaesthesia. PMID- 15795968 TI - Thoracolumbar kyphosing scoliosis associated with spondyloepiphyseal dysplasia congenita: a case report. AB - BACKGROUND CONTEXT: A case of spondyloepiphyseal dysplasia congenita (SEDC) with thoracolumbar kyphosing scoliosis and a clinical outcome of the patient's surgical treatment are reported. PURPOSE: We report a rare case and the effect of surgical treatment on the kyphosing scoliosis with SEDC with a review of literature. STUDY DESIGN/SETTING: SEDC is a rare disease and has a variety of spinal deformities. To our knowledge, a case of surgical treatment for the kyphosing scoliosis with SEDC was not reported before. METHODS: The patient, a 27 year-old woman, complained of severe back pain and dyspnea. She was operated on in 1997 for severe kyphosing scoliosis, using segmental spinal instrumentation and strut bypass graft. She was followed for 6 years, and clinical symptoms and plain X-ray films were investigated. RESULTS: Her kyphosis was corrected from 116 degrees to 86 degrees at the final follow-up; otherwise, her scoliosis was almost unchanged. Her symptoms were relieved. CONCLUSIONS: A case of SEDC with thoracolumbar kyphosing scoliosis was successfully treated by segmental spinal instrumentation and anterior strut bypass graft. PMID- 15795969 TI - From advanced imaging services... Where the source of postoperative headache and radiculopathy is revealed. PMID- 15795971 TI - [Modern approach to examination and operative treatment of patients with considerable atrophy of jaws]. PMID- 15795970 TI - From advanced imaging... When incidental findings masquerade as spinal pathology. PMID- 15795972 TI - [Experience in ablation of skin surplus and lipocele in the region of lower eyelids]. PMID- 15795973 TI - [Content of low- and medium-molecular weight substances in biological substrates in cases of cheilitis in children and teenagers]. PMID- 15795974 TI - [Factors of specific and non-specific defense in pathogenesis of early forms of periodontal disease]. PMID- 15795975 TI - [Experience in light-science teaching on courses of post-graduate education of stomatologists]. PMID- 15795976 TI - Re: Lateral rotation mattresses for wound healing. PMID- 15795977 TI - Robots and humans: synergy in planetary exploration. AB - How will humans and robots cooperate in future planetary exploration? Are humans and robots fundamentally separate modes of exploration, or can humans and robots work together to synergistically explore the solar system? It is proposed that humans and robots can work together in exploring the planets by use of telerobotic operation to expand the function and usefulness of human explorers, and to extend the range of human exploration to hostile environments. PMID- 15795978 TI - Paging Nancy Chatham. PMID- 15795979 TI - Safety analysis of contained low-hazard biotechnology applications. AB - A technical safety analysis has been performed on a containment-level-2 pilot plant in order to assess an upgrading of the existing facility, which should comply with good manufacturing practices. The results were obtained by employing the hazard and operability (HAZOP) assessment method and are discussed in the light of the appropriateness of this procedural tool for low-hazard biotechnology applications. The potential release of micro-organisms accounts only for a minor part of the hazardous consequences. However, in certain cases the release of a large or moderate amount of micro-organisms would not be immediately identified. Most of the actions required to avoid these consequences fall into the realm of operational procedures. As a major part of potential failures result from human errors, standard operating procedures play a prominent role when establishing the concept of safety management. The HAZOP assessment method was found to be adequate for the type of process under investigation. The results also may be used for the generation of checklists which, in most cases, are sufficient for routine safety assurance. PMID- 15795980 TI - Healthcare Commission calls for help as complaints flood in. AB - The NHS's new complaints system is buckling under massive pressure. The Healthcare Commission is facing a backlog of more than 3,700 grievances and a private company will be brought in next week to help clear the build-up. The commission expects to receive 9,000 complaints by the end of its first year- almost double the number anticipated. PMID- 15795981 TI - On long-term conditions. PMID- 15795982 TI - On building local partnerships. PMID- 15795983 TI - Volunteering. give and take. PMID- 15795984 TI - Recruitment. Finance staff don't grow on trees. PMID- 15795985 TI - More Americans willing to limit physician-hospital choice for lower medical costs. AB - More Americans are willing to limit their choice of physicians and hospitals to save on out-of-pocket medical costs, according to a new national study by the Center for Studying Health System Change (HSC). Between 2001 and 2003, the proportion of working-age Americans with employer coverage willing to trade broad choice of providers for lower costs increased from 55 percent to 59 percent- after the rate had been stable since 1997. While low-income consumers were most willing to give up provider choice in return for lower costs, even higher-income Americans reported a significant increase in willingness to limit choice. Compared with other adults, people with chronic conditions were only slightly less willing to limit their choice of physicians and hospitals to save on costs. Perhaps as a result of growing out-of-pocket medical expenses in recent years, the proportion of people with chronic conditions willing to trade provider choice for lower costs rose substantially from 51 percent in 2001 to 56 percent in 2003. PMID- 15795986 TI - [Autoimmune hepatitis]. PMID- 15795987 TI - The impact of Medicare home health policy changes on Medicare beneficiaries: part II. AB - This brief presents the results of a follow-up study (Murtaugh et al., 2003) on the effects of the Balanced Budget Act of 1997 on Medicare home health service use and beneficiary outcomes.[The results of the initial study (McCall et al., 2001) were discussed in a Spring 2003 policy brief and fact sheet.] The 2003 study found that the new payment systems have had a significant impact on the Medicare home health benefit: utilization declined, aggregate payments and payments per visit decreased (and then increased), the mix of services shifted, and the types of patients served appears to have changed. These results show that policy goals-in this case attempting to limit the use of the Medicare home health benefit while shifting services towards skilled care-can be instituted through changes in the payment system, though it is important to examine the impact of changes for possible unintended effects. Further study on the sustained impact of the current payment system-particularly on quality of care-is still needed. PMID- 15795988 TI - [Eosinophilic hepatic abscess]. PMID- 15795989 TI - [Ultrastructure of chronic liver diseases--lysosomes and lysosomal storage diseases]. PMID- 15795990 TI - Managed care and long-term care: a potential solution? AB - This brief summarizes research by Sparer (2003) and Hughes (2002) on managed care issues for people with long-term care needs. Managed care has been proposed as a solution to the difficult problem of managing the health of this complex population. Sparer's research examines the reasons behind the failure of states' attempts to encourage commercial managed care plans to take on this task. He argues that understanding these reasons can yield important lessons for the mostly provider-sponsored plans that currently serve the long-term care population, as well as for states, that aim to save money and better serve this population. Hughes' evaluation of a small, provider-sponsored organization found that when the organization became a managed care provider, it faced significant organizational challenges that forced it to become more efficient. Importantly, it showed that the use of capitation reimbursed the plan more effectively than fee for service, increased the range of services provided to clients, and maintained client satisfaction; however, capitation did not affect the rate of nursing home admission. PMID- 15795991 TI - Creating more choices for older adults: financing LTC through an integrated income and disability annuity. PMID- 15795992 TI - In memoriam Yasutomi Nishizuka 1932-2004. PMID- 15795993 TI - Cholinesterase inhibitory and spasmolytic potential of steroidal alkaloids. AB - A new steroidal alkaloid, isosarcodine (1) along with four known bases, sarcorine (2), sarcodine (3), sarcocine (4) and alkaloid-C (5) were isolated from the MeOH extract of Sarcococca saligna. The structures of these alkaloids were identified by spectral data interpretation. These compounds were subjected to acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition studies, and were found to be noncompetitive inhibitors of AChE (Ki = 21.8, 90.3, 32.2, 16.0 and 50.0 microM, respectively) and uncompetitive or noncompetitive inhibitors of BChE (Ki = 8.3, 7.5, 15.6, 5.0 and 12.0 microM, respectively). The compounds (2-5) also showed dose-dependent spasmolytic activity in the rabbit jejunum intestinal preparations and also relaxed the high K+ (80 mM)-induced contraction, indicative of a calcium channel-blocking mechanism. Structure activity relationship suggested that the nitrogen substituents at C-3 and/or C-20 of steroidal skeleton and the hydrophobic properties of the pregnane skeleton are the key structural features contributed to the inhibitory potency of these steroidal alkaloids against AChE and BChE. PMID- 15795994 TI - Clinical guidelines and their civil law effects. AB - The number of clinical guidelines or protocols for medical treatment is growing. Considered as an important instrument for increasing the quality of medicine, these originally intended technical guidelines turn out to influence doctor's civil liability. Ignoring those guidelines in itself may cause a breach of a doctor's duty of care according to civil law and he may be charged to produce evidence that his medical treatment was correct. This development is taking place in several countries as France, Germany, United Kingdom and the Netherlands. PMID- 15795996 TI - The Norwegian system of compensation to patients. AB - In Norway we have come to the conclusion that a traditional compensation law based on negligence is not the right "tool" for handling difficult questions as to whether or not a patient shall get compensation if required after medical treatment or other forms of contact with the medical profession. The compensation scheme established in 1988 involving the public sector of the Norwegian health care system has received almost 18000 claims since Jan 1 1988, and has paid out more than 1.2 billion NOK (137 million US$) in compensation during these years. At the same time, the number of compensation claims brought before the courts has decreased. After 13 years of a provisional scheme dealing with patient injury compensation based on no-fault rules, we find that compensation claims indeed can be handled without focus on blame and negligence, and that this ensures both that the patients get equitable compensation and that unnecessary legal procedures are avoided. The system also has had a favourable impact on interaction within the health care sector and upon relationships between health care personnel and patients. PMID- 15795995 TI - Physicians and genetic malpractice. AB - Primary care physicians are unprepared for the increase in demands for prenatal genetic testing. Often, they do not possess the necessary knowledge, skills or attitudes to provide genetic counselling. Yet, since the demand for prenatal genetic services is growing faster than the number of genetic professionals, the responsibility of genetic counselling will fall to these physicians. Physicians who lack genetic literacy may find themselves the targets of lawsuits for wrongful birth and wrongful life. Wrongful birth and wrongful life claims (in the context of genetics) both assert that but for the physician's negligence, the handicapped child would not have been born. Such medical malpractice suits against physicians exist in the United States, the United Kingdom, Canada and Australia. This paper discusses the case law on wrongful birth/life cases in these four countries. The authors conclude that as the number and availability of prenatal genetic tests increases, so too will the number of genetic malpractice claims, unless the education of physicians and medical students in genetics is promoted, possibly with the Internet as the new educational paradigm. PMID- 15795997 TI - Wagging the watchdog: law and the emergence of bioethical norms. AB - Bioethics offers an ideal vantage point to study how law and ethics affect the emergence of norms. Moral philosophy traditionally places itself in the role of prime mover, the normative force behind social, cultural and legislative change. But some bioethical norms, particularly those associated with emerging right-to die norms in the mid 1960's and through the late 1980's, did not originate in this way. Instead, legal reasoning and judicial decision-making brought about rapid change in bioethical norms almost excluding moral philosophy in the process. Judicial decision-making has prompted legislation, molded public opinion, guided medical practice and shaped moral thinking. The reasons for this are not difficult to understand. Legal reasoning, usually in the form of risk management, replaces moral reasoning because it is cognitively more comfortable, rational and parsimonious. This process creates undue pressure on the judiciary to undertake tasks for which it may not be well trained, while at the same time offers a challenge to ethicists to advocate interdisciplinary, deliberative and public forums to attenuate the undue influence of the law. PMID- 15795998 TI - Developments in patients' rights in Turkey. AB - Today, the social, economic, cultural, ethical, and political climate has given rise to an international movement, that is re-evaluating the importance of patients' rights. In Turkey, as competition continues to increase between public and private hospitals, the emphasis on presenting health care services in the most advantageous way has become a focal point of debate. As a result, patients' rights issues are being actively discussed in both the private and public health care sectors in the country. In this study, the development of patients' rights issues in Turkey is reviewed and the results of some selected descriptive studies are summarized. Suggestions are made toward policies for effective and efficient health care provision in terms of both broad and specific issues that have arisen in the field of patients' rights. PMID- 15795999 TI - The development and implementation of patients' rights: Dutch experience of the right to information. AB - Many initiatives have been taken to advocate, develop and emphasize patients' rights. The existence of legislation, case law or charters in the area of patients' rights does not guarantee that these rights are or will be successfully implemented in everyday practice. The implementation of patients' rights requires specific actions and expertise. It is important to develop strategies to enhance the successful implementation of patients' rights. This is illustrated by using the example of the developments in the Netherlands regarding the patient's right to information. This development shows that legal interventions, such as legislation, will always have to be embedded in and/or supplemented by non-legal policy measures. This calls for a broad and well-considered implementation policy, including items at various levels (legislation, patient and patient organizations, health providers and health institutions, contextual conditions). Such a strategy calls for a multidisciplinary approach, involving input from the areas of law, ethics, medicine, the sciences etc. PMID- 15796000 TI - The abuse of children in Greece. AB - Children are being abused in Greece at an alarming rate. This article aims to clarify the magnitude of the child abuse issue in Greece and to highlight medical and legal aspects that may play a contributory role in the existing defects in the revelation, prevention, and punishment of this type of crime. PMID- 15796001 TI - The Bioethics Convention of the Council of Europe and organ sharing for transplant recipients in Scandinavia. AB - Even though organ transplantation is often life saving, the lack of donor organs is limiting the number of transplantation procedures. In small countries, like the Scandinavian countries, the small population level highlights this problem of organ availability which is further complicated by the fact that the utilisation of available organs may be prevented by histoincompatibility between the host and donor. This problem can only be solved by sharing both medical information and organs across countries. In Scandinavia, an organ sharing program (Scandiatransplant) was initiated between Denmark, Finland, Iceland, Norway and Sweden in the late 1960's long before the establishment of European Economic Area and the expansion of the European Union in Northern Europe. Even though trade in human organs is prohibited by international conventions, medical procedures and services that are associated with transplant activities are such services whose "free movement" within the Union is guaranteed by the Convention of Rome. These services can thus be offered across the national borders for remuneration. The potential impact of the conventions of the Council of Europe on transplantation services and organ sharing programs within European Union will be discussed. PMID- 15796002 TI - Epilepsy--a legal perspective on obligations owed to patients by medical practitioners. PMID- 15796003 TI - Epilepsy and the doctor's duty of care: ethical and legal issues. PMID- 15796004 TI - External injury marks (wounds) on the head in different types of blunt trauma in an autopsy series. AB - The localization, length and number of deep scalp wounds caused by falls on a flat surface (n = 203), falls down stairs (n = 51) and from blows (n = 51) were examined in a comparative analysis of 305 deaths. This confirmed the common finding in forensic literature that head injuries from blows occur more often (55% of cases) above the so-called hat-brim line (= greatest horizontal head circumference) than injuries from falls on a flat surface. On the other hand, around one third of the wounds caused by falls were located above this line. In blows and falls down stairs, but not in falls on a flat surface, the wounds close to the crown of the head were markedly longer than those in or below the hat-brim line. Regarding wounds inside the hat-brim line, there were hardly any differences between the three trauma types. PMID- 15796005 TI - Women undergoing investigation of sexual abuse in the metropolitan area of Porto Alegre, Brazil: a retrospective study. AB - This is a retrospective study of 1,063 examinations performed on women at the Institute for Forensic Medicine in the city of Porto Alegre, RS, Brazil, between January 1998 and December 1999, as part of an investigation of charges of sexual abuse. Most women were white (83.9% - 892), single (89.1% - 947), and were between 11 and 20 years of age (59% - 627); 40% (425) were 14 years old or younger. There was evidence of previous sexual contact in 57% (606) of the patients; 37% (393) were virgins. Evidence of recent sexual intercourse was found in 287 (27%) cases, including presence of sperm in the vagina (18% - 191), recent rupture of the hymen (7% - 74), and pregnancy (2% - 21). Physical violence was evident in 30% (319) of all cases. In cases where information regarding the perpetrator was available (39% - 415), 25% (105) consisted of family members. Further studies should be carried out in order to assess the incidence of sexual violence by family members. PMID- 15796006 TI - Reports of medical experts in cases of drug addiction and assessment of mitigating circumstances by the court. AB - In Spain, as the law stands, an accused person wishing to cite drug addiction as a mitigating circumstance in sentencing must demonstrate this addiction. Expert evidence therefore acquires great importance for an eventual sentence. The aim of this study is to analyse the sentencing process in cases in which the relevance of drug addiction as a mitigating circumstance in criminal responsibility is sought. Of a total of 261 sentences handed down in the Criminal Court of Murcia, we studied 139 sentences that cited drug addiction as one of the mitigating circumstances in criminal responsibility. A forensic report was presented in only 36 cases (25.9%) and in most of these cases (61.1%) the individual's drug addiction was assumed from observing signs of recent venous punctures. There were no objective data (blood and urine analysis) in 128 cases (92.1%). However, mitigating circumstances as regards criminal responsibility were considered and accepted by the Court in 130 cases (93.4%). We should emphasise the limited, not to say total absence of, clinical data in the submitted reports, which implies the impossibility of scientifically testing the disputed circumstances. PMID- 15796007 TI - Malarial (Plasmodium falciparum) dihydrofolate reductase-thymidylate synthase: structural basis for antifolate resistance and development of effective inhibitors. AB - Dihydrofolate reductase-thymidylate synthase (DHFR-TS) from Plasmodium falciparum, a validated target for antifolate antimalarials, is a dimeric enzyme with interdomain interactions significantly mediated by the junction region as well as the Plasmodium-specific additional sequences (inserts) in the DHFR domain. The X-ray structures of both the wild-type and mutant enzymes associated with drug resistance, in complex with either a drug which lost, or which still retains, effectiveness for the mutants, reveal features which explain the basis of drug resistance resulting from mutations around the active site. Binding of rigid inhibitors like pyrimethamine and cycloguanil to the enzyme active site is affected by steric conflict with the side-chains of mutated residues 108 and 16, as well as by changes in the main chain configuration. The role of important residues on binding of inhibitors and substrates was further elucidated by site directed and random mutagenesis studies. Guided by the active site structure and modes of inhibitor binding, new inhibitors with high affinity against both wild type and mutant enzymes have been designed and synthesized, some of which have very potent anti-malarial activities against drug-resistant P. falciparum bearing the mutant enzymes. PMID- 15796008 TI - Avian malaria in Brazilian passerine birds: parasitism detected by nested PCR using DNA from stained blood smears. AB - The microscopical examination of Giemsa-stained thin blood smears and a nested PCR were performed to detect avian Plasmodium in 275 passerine birds from small and large fragments of Atlantic Forest, Minas Gerais, Brazil. The 275 blood smears were used both for the microscopical examination and nested PCR providing the DNA template used for the reactions. The sensitivity of the nested PCR assay was higher than that observed for blood smears through microscopical examination. High prevalence (39.6%) of Plasmodium infections was detected by nested PCR while the microscopical examination detected only 16.5 % positive birds. Poor agreement was observed between the results of the two different tests. The PCR data obtained were correlated to the forest fragment size of the Atlantic Forest and also correlated to the biological characteristics of the birds (nest type construction, diet, participation in mixed-species flocks, age and sex). Birds captured in the large forest areas were more infected than birds captured in the small areas (51.9 % and 28.5 %, respectively). Diet and participation in mixed species flocks were correlated to the Plasmodium parasitism. The insectivorous birds and those that participated in mixed-species flocks were more frequently infected (47% and 41.5%, respectively) than the other groups. PMID- 15796009 TI - In vivo killing of Giardia trophozoites harbouring bacterial endosymbionts by intestinal Paneth cells: an ultrastructural study. AB - To date Paneth cells have not previously been reported to kill Giardia trophozoites and other protozoa in vivo. Here we report the first evidence for in vivo killing of Giardia trophozoites by intestinal Paneth cells. Transmission electron microscopic (TEM) examination of duodenal specimens taken from naturally infected mice revealed that only Giardia trophozoites harbouring peripheral bacterial endosymbionts were destroyed and lysed in the vicinity of the activated Paneth cells. Additionally, intestinal epithelium was more affected by Giardia harbouring bacterial endosymbionts than Giardia with no endosymbionts. Our findings imply that the bacterial endosymbionts within Giardia trophozoites have a role in both host protective and pathological mechanisms, probably through altering the trophozoite antigencity. These observations might shed light on the diversity in infectivity and host specificity of Giardia species. PMID- 15796010 TI - Identification of the first pyrimidine nucleobase transporter in Leishmania: similarities with the Trypanosoma brucei U1 transporter and antileishmanial activity of uracil analogues. AB - While purine transport has been widely studied in protozoa, almost nothing is known about their capacity to salvage pyrimidines. Here, we report a Leishmania major transporter with high affinity for uracil (Km=0.32+/-0.07 microM) which we designated LmU1. This transporter displayed a high degree of specificity, as it had virtually no affinity for cytosine, thymine or purine nucleobases, nor did it transport pyrimidine nucleosides. Highest affinity was for 5-fluorouracil. The results show that the permeant binding site of LmU1 interacts strongly with the keto groups of uracil, as shown by a low affinity for 2-thio- and 4-thiouracil. LmU1 appears to further bind uracil through a weak hydrogen bond with N(1)H of the pyrimidine ring in addition to a stronger H-bond with N(3)H. Substrate binding and selectivity were strikingly similar to that of the U1 transporter in the related kinetoplastid Trypanosoma brucei. Uracil analogues likely to be transported by LmU1 were also screened for antileishmanial activity, with 5 fluorouracil displaying strong activity against promastigotes and intracellular amastigotes. Overall, the results show that, like purine nucleobase transport, pyrimidine nucleobase transport function is very similar in L. major and T. brucei insect forms. PMID- 15796011 TI - Cell invasion and intracellular fate of Encephalitozoon cuniculi (Microsporidia). AB - Microsporidia are obligate intracellular parasites that utilize a unique mechanism to infect host cells, which is one of the most sophisticated infection mechanisms in biology. Microsporidian spores contain a long coiled polar tube that extrudes from the spores and penetrates the membranes of new host cells. We have initiated a study to investigate the invasive process and intracellular fate of the microsporidium Encephalitozoon cuniculi. Here we show that relatively few cells were infected through the traditional penetration of the polar tube from outside. Rather, phagocytosis of spores occurred at least 10 times more frequently than injection of sporoplasms. Some spores extruded their polar tube inside the cells following phagocytosis. Membranes of the vacuoles surrounding the internalized spores were positive for late endosomal and lysosomal markers. Spores that remained inside these compartments disappeared within 3 days. Thus, our studies demonstrate that in addition to the unique way in which microsporidia infect host cells, E. cuniculi spores can also gain access to host cells by phagocytosis. The presence of intracellular spores that have extruded their polar tube shows that some spores germinate after phagocytosis, thus escaping from the phagosomes that mature into lysosomes. PMID- 15796012 TI - Toxoplasmosis in Pallas' cats (Otocolobus manul) raised in captivity. AB - Manuls or Pallas' cats (Felis manul, syn. Otocolobus manul) are endangered wild cats from Central Asia kept and bred in many zoos. Despite good breeding success young cats frequently die from acute toxoplasmosis. From 1998 to 2002, a breeding pair in the Schonbrunn Zoo in Vienna, Austria, gave birth to 24 kittens; 58 % of kittens died between the 2nd and the 14th week of life, mostly due to acute toxoplasmosis. The epidemiology of toxoplasmosis in Pallas' cats was examined and a control strategy to protect the kittens from fatal toxoplasmosis was developed. One 12-week-old kitten from a litter of 6 born in 2001 died of generalized toxoplasmosis. This kitten had shed T. gondii oocysts that were bioassayed in mice. Toxoplasma gondii was isolated in tissue culture inoculated with tissues of these mice. The surviving animals were immediately treated with clindamycin for 16 weeks; they acquired a natural infection and seroconverted by the end of this time without clinical signs. PMID- 15796013 TI - High levels of congenital transmission of Toxoplasma gondii in longitudinal and cross-sectional studies on sheep farms provides evidence of vertical transmission in ovine hosts. AB - Recent research suggests that vertical transmission may play an important role in sustaining Toxoplasma gondii infection in some species. We report here that congenital transmission occurs at consistently high levels in pedigree Charollais and outbred sheep flocks sampled over a 3-year period. Overall rates of transmission per pregnancy determined by PCR based diagnosis, were consistent over time in a commercial sheep flock (69%) and in sympatric (60%) and allopatric (41%) populations of Charollais sheep. The result of this was that 53.7 % of lambs were acquiring an infection prior to birth: 46.4% of live lambs and 90.0% of dead lambs (in agreement with the association made between T. gondii and abortion). No significant differences were observed between lamb sexes. Although we cannot distinguish between congenital transmission occurring due to primary infection at pregnancy or reactivation of chronic infection during pregnancy, our observations of consistently high levels of congenital transmission over successive lambings favour the latter. PMID- 15796014 TI - The role of acidic organelles in the development of schistosomula of Schistosoma mansoni and their response to signalling molecules. AB - The cercariae of Schistosoma mansoni become transformed into schistosomula during host skin penetration. We have found that large acidophilic compartments are detected in schistosomula but not in cercariae or in any other stages of the parasite by use of the fluorescent dye LysoTracker, a dye specific for mammalian lysosomes. Some of these large acidic compartments incorporated monodansylcadaverine, a specific dye for autophagosomes. We have used potent inhibitors (wortmannin and 3-methyladenine) and a potent inducer (starvation) of autophagy to show that the pathway to the formation of the acidic compartments requires specific molecular signals from the environment and from the genome. Certain doses of ultraviolet light inhibited significantly the formation of the acidic compartments, which may indicate disruption of the lysosome/autophagosome pathway. We have also defined two proteins that are commonly associated with lysosomes and autophagosomes in mammalian cells, the microtubule-associated membrane protein (MAP-LC3) and lysosome-associated membrane protein (LAMP-1), in extracts of schistosomula. We suggest that the autophagy pathway could be developed in transformed schistosomula. PMID- 15796015 TI - Spread of parasites in metapopulations: an experimental study of the effects of host migration rate and local host population size. AB - We established experimental metapopulations of the flour beetle, Tribolium castaneum, and its ectoparasitic mite, Acarophenax tribolii, to investigate the effects of host migration rate and local host population size on the spread of mite infections. Global prevalence across our metapopulations was less than half the observed within-patch prevalence, so that spatial structure alone afforded a great deal of protection to hosts against parasite infection. Our results showed further that migration played a determining role in occupancy, the number of patches infected within a metapopulation, while host population size played a determining role in local prevalence, the fraction of hosts infected within local patches. Local and global prevalence appeared to reach equilibrium levels on 2 different time-scales. Local host prevalence reached equilibrium values within 30 days of receiving an infected host migrant. Global prevalence increased more slowly and was clearly dependent upon occupancy, the number of host patches with at least 1 infected host, which in turn depended on the level of host migration among host patches. The effect of population size was not limited to local prevalence in patches without spatial structure but extended to sets of patches across the metapopulation. Lloyd's index of patchiness differed significantly between metapopulations with small versus large numbers of hosts. Although parasites were aggregated on hosts for both local patch sizes, they tended to aggregate to a much greater degree at the smaller host patch size. We discuss our empirical findings in light of current epidemiological theory. PMID- 15796016 TI - Manipulation of host cytokine network by ticks: a potential gateway for pathogen transmission. AB - Ticks are obligatory blood-feeding arthropods that secrete various immunomodulatory molecules to antagonize host inflammatory and immune responses. Cytokines play an important role in regulating these responses. We investigated the extent to which ticks interact with the sophisticated cytokine network by comparing the effect of salivary gland extracts (SGE) of 3 ixodid tick species, Dermacentor reticulatus, Amblyomma variegatum and Ixodes ricinus, all of which are important vectors of tick-borne pathogens. Using specific ELISAs, anti cytokine activity was demonstrated with 7 cytokines: IL-8, MCP-1, MIP-1alpha, RANTES, eotaxin, IL-2 and IL-4. The results varied between species, and between adult males and females of the same species. Relatively high activity levels were detected in saliva of female D. reticulatus, confirming that the observed anti cytokine activities are an integral part of tick saliva secreted into the host. Results with fractionated SGE indicated that from 2 to 6 putative cytokine binding molecules are produced, depending on species and sex. Binding ability of SGE molecules was verified by cross-linking with radio-isotope labelled MIP 1alpha. By targeting different cytokines, ixodid ticks can manipulate the cytokine network, which will greatly facilitate blood-feeding and provide a gateway for tick-borne pathogens that helps explain why ticks are such efficient and effective disease vectors. PMID- 15796017 TI - Efficacy of two cyclooctadepsipeptides, PF1022A and emodepside, against anthelmintic-resistant nematodes in sheep and cattle. AB - Resistance against the major currently available anthelmintics has reached a critical level in many small ruminant herds world-wide, and is increasingly becoming a problem in horses and cattle. Therefore, new products with different modes of action are urgently needed. Recently, such a new class of compounds, the anthelmintically active cyclooctadepsipeptides, was described. Here, the effects of cyclooctadepsipeptides on benzimidazole-, levamisole- and ivermectin-resistant populations of Haemonchus contortus in sheep as well as an ivermectin-resistant Cooperia oncophora population in cattle were studied. Experimentally infected sheep and cattle were used. Animals were treated orally, subcutaneously, or intravenously with cyclooctadepsipeptides. The anthelmintic effects were assessed by means of fecal egg count reductions and/or worm count reductions. Both, PF1022A and emodepside were found to be fully effective against these parasite populations. These findings confirm that this new class of compounds acts by a different mode of action compared to the above-mentioned anthelmintics. PMID- 15796018 TI - Serpin expression in the parasitic stages of Trichostrongylus vitrinus, an ovine intestinal nematode. AB - Members of the genus Trichostrongylus, such as T. vitrinus, being endemic in Northern Europe, are among the principal causative nematodes which contribute to parasitic gastro-enteritis in sheep world-wide, inhabiting the proximal small intestine and causing damage to the mucosa. This results in impaired nutrient absorption as well as a pronounced inflammatory response with cellular infiltration of the mucosa, including a pronounced mast cell response. These mast cells release serine proteinases that enhance the passage of effector cells and macromolecules across epithelial boundaries and into direct contact with the invading parasite. The adult and larval stages of T. vitrinus release a number of serine proteinases in vitro that may contribute to tissue invasion and nutrient acquisition in vivo. This study describes the molecular cloning and characterization of a serine proteinase inhibitor (serpin) that is present in extracts of all the parasitic stages, becoming more abundant as the life-cycle progresses. The serpin is present in the in vitro excretory/secretory products (ES) of 4th-stage larval and adult parasites, being more abundant in the former. The serpin was expressed in E. coli and the recombinant protein was a potent inhibitor of several host serine proteinases including mast cell proteinases. The serpin may regulate the activity of the parasite serine proteinases or it may modulate the host immune response to the parasite by inhibiting the activity of serine proteinases released from host inflammatory cells. PMID- 15796019 TI - Phylogeny, evolution and host-parasite relationships of the order Proteocephalidea (Eucestoda) as revealed by combined analysis and secondary structure characters. AB - In a manner similar to many other groups of organisms, the tapeworm order Proteocephalidea poses a difficult phylogenetic problem if treated on the basis of single-gene analysis. Since the biogeography and host distribution of proteocephalideans make these tapeworms a potentially interesting model for evolutionary and co-evolutionary studies, we tried to resolve their phylogenetic relationships by applying a multi-gene approach. The ITS2 sequences and V4 hypervariable loop of 18S rRNA were obtained for 43 and 35 proteocephalidean taxa, respectively, and combined with other sequences available in the GenBank. The phylogenetic analysis of the combined DNA set was confronted with characters derived from ITS2 secondary structures. Using this approach, a species-rich Neotropical lineage of proteocephalideans could be reliably resolved. The phylogenetic relationships within this group show a high degree of phylogeny independent host distribution. The reconstruction of ITS2 secondary structure revealed a universal 4-domain arrangement, which is conserved across a wide range of Neodermata. Several motifs of the secondary structure could be mapped to the phylogenetic tree as possible clade synapomorphies. PMID- 15796020 TI - Information(ist) please? PMID- 15796021 TI - Electronic fund transfer system--making your ILL life easier. PMID- 15796023 TI - Collection development post-Bandon/Hill: what to do? PMID- 15796024 TI - Hospital library increases space, staff and budget. PMID- 15796022 TI - EFTS saves time and money for hospitals. PMID- 15796025 TI - [Physiological data analysis using Igor Pro: the data display and analysis]]. PMID- 15796026 TI - [Rapidly increasing allergic diseases]. AB - It is well known that allergic diseases, such as allergic rhinitis, asthma, atopic dermatitis, and food allergy, are increasing in prevalence. Epidemiological studies show that the prevalence of allergic diseases is approximately 40% in children and 30% in adult. Thus allergic diseases are global health problem in Japan. Changes in environment or lifestyle may be one of the causes of the increase in allergic diseases. Therefore it may be a great concern for many people how they prevent the diseases or how they manage allergic symptoms. The major purpose of this citizen joint symposium was to broaden citizen's knowledge on the mechanism, epidemiology, and appropriate management of allergic diseases. We hope that this symposium provided useful information to the participants about the current trends in the management of allergic diseases. PMID- 15796027 TI - [Gene expression profile of infiltrating pancreatic duct cancer by using cDNA array]. PMID- 15796029 TI - [Changes in gene expression profile associated with tumor proliferation in esophageal cancer]. PMID- 15796030 TI - [Gene expression analysis using cDNA array in infiltrating ductal carcinoma]. PMID- 15796028 TI - [Study on perineural infiltration in bile duct neoplasm--with special reference to the relationship with RET]. PMID- 15796031 TI - [Elaboration of newly discovered functions of budding yeast homolog Sla2p of protein HIP1 (Huntington-interacting protein 1) that binds huntingtin, a Huntington-causing protein]. PMID- 15796032 TI - [Study on the correlation between vascular complications suffered by diabetic patients and nick beta-2GPI ratio]. PMID- 15796033 TI - [A comparison of anomalies in the skeletal muscle energy metabolism of the upper and lower extremities during exercise by patients with chronic heart failure]. PMID- 15796034 TI - [Augmented expression of endoplasmic reticulum oxidoreductin 1-like (ERO1-L) under a hypoxic condition]. PMID- 15796035 TI - [Immunohistochemical study on proliferation and prognostic factors of bile duct neoplasms at the portal section --with special reference to bax protein expression]. PMID- 15796036 TI - [Study on the Wnt/beta-catenin pathway in autosomal dominant polycystic kidney disease--a cytobiological evaluation of pkd1 gene]. PMID- 15796037 TI - [Vascular endothelial growth factor (VEGF) production by a dopamine D2 receptor agonist and its action mechanism in glioma cells]. PMID- 15796038 TI - [Study on the suppressive effect of prostaglandin E1 on endothelial hypertrophy of an autologous venous graft]. PMID- 15796039 TI - [Clinical and pre-clinical studies on local hemodynamics of esophagogastric varices determined by endoscopic microvascular Doppler sonography]. PMID- 15796040 TI - [Neuron-protective effect of ischemic preconditioning in a rabbit reperfusion model following spinal ischemia]. PMID- 15796041 TI - [Immunotherapy combining leukemic cell-derived shock protein and dendritic cells applied to leukemic mice following homologous bone marrow transplantation]. PMID- 15796042 TI - [Sequential changes in HGF, TGF-beta1 and c-Met protein expression in chronically over-extended bladder]. PMID- 15796044 TI - [Thermal imaging properties of toes after walking stress test in diabetic patients]. AB - In consecutive 50 diabetic patients hospitalized for medical education, without subjective symptoms of autonomic neuropathy (DM group), performed treadmill walking stress test and thermography, for clarify the feature of the pattern typical of diabetic autonomic neuropathy. Thermal images were collected, before, immediately after, 3, 6, 12 minutes after walking. The mean temperature of toes fell more than 1 degree C than that of baseline level and returned within 0.5 degrees C of baseline level within 6 minutes (N type) in 66% of 30 normal subjects (C group) and 24% of DM group. In 10% of C and 24% of DM, the temperature fell but not returned within 0.5 degrees C of baseline level in 6 minutes (D type). In 17% of C and 38% of DM, the temperature changed within 0.5 degrees C (F type), or rose more than 0.5 degrees C after exercise (U type) in 7% in C and 14% in DM groups. Pts D, F group, have more complications (HbA1c, nephropathy, retinopathy or somatic neuropathy), but not so in C, U type. We concluded D, F types were the typical thermographic features of the toes of pts with diabetic autonomic neuropathy. PMID- 15796043 TI - [Evaluation of novel test strip to measure albumin and creatinine in urine]. AB - The purpose of this study was to evaluate the clinical usefulness of novel test strip that simultaneously measure urinary albumin and creatinine. Testing was performed on 95 random urine samples from diabetics. Each sample was assayed with following methods: test strip by instrument (AX4280) reading and by visual interpretation, quantitative method for albumin, creatinine, and alpha1 microglobulin and cellulose acetate membrane electrophoresis. The results of test strip had good correlation with quantitative results. In the case of instrument reading, the sensitivity, specificity and consistency were 91.2%, 78.9% and 86.3% for albumin and 94.5%, 87.5% and 91.6% for albumin index (albumin/creatinine ratio; A/C ratio), respectively. The percent same level agreement for creatinine was 63.2%. After correction using creatinine value on test strip, 18.4% of samples defined as negative by albumin quantitative value were turned to be positive, 12.3% of samples defined as positive were turned to be negative. The same level of creatinine correction effect as the quantitative method was obtained at the test strip. When alpha1-microglobulin/creatinine (alpha1 m/g x Cr) ratio was compared with albumin index, percent positive of alpha1 m/g x Cr ratio was 35.0% for samples with albumin index less than 30 mg/g x Cr, 59.5% for those with albumin index between 30 and 300 mg/g x Cr. In addition, on cellulose acetate membrane electrophoresis, both retinol binding protein (RBP) and beta2 microglobulin (beta2 m) were detected in 12.5% of patients in normal condition or pre-nephropathy. Among patients at early-nephropathy, the detection rate of RBP and beta2 m were 20.0% and 15.0% respectively. These results indicate that the renal tubule is also damaged at the early stages of nephropathy. PMID- 15796045 TI - [Urinary diacetylspermine: its analysis and performance as a novel tumor marker]. AB - N1,N12-Diacetylspermine (DiAcSpm) is excreted in the urine of healthy persons as a minor component of urinary polyamine. It is a promising tumor marker, since its excretion is frequently elevated in patients with various types of cancers. DiAcSpm was first detected and characterized by HPLC fractionation followed by enzymatic detection, but more recently, antibodies highly specific for DiAcSpm was prepared, and an ELISA system applicable to determination of urinary DiAcSpm was established. Measurement of urinary DiAcSpm using this ELISA system revealed that DiAcSpm is able to detect early stage (m and sm) colon cancers which CEA and CA19-9 cannot detect. DiAcSpm may also serve as a prognostic indicator and a marker for recurrence of colon cancer. Urinary DiAcSpm is elevated in metastatic and primary brain tumors including grade 3 and 4 gliomas and primary central nervous system lymphoma. In these primary brain tumors changes in urinary DiAcSpm were well correlated with the efficacy of treatments, recurrence of disease and increased malignancy of a tumor. DiAcSpm may be useful as a comprehensive indicator of the activeness of a brain tumor lesion in a patient. DiAcSpm was elevated in hepatocellular carcinoma, but patients with liver cirrhosis also showed considerably elevated levels of DiAcSpm. PMID- 15796046 TI - [Urine diacetylspermine as a novel tumor marker]. AB - A urine tumor marker, diacetylspermine, was examined in patients with recurrent pancreato-biliary carcinoma, liver tumor, lung carcinoma and gynecologic malignancies. The urine marker increased together with recurrence, suggesting a recurrence monitoring marker at the outpatient ward. Regarding hepatocellular carcinoma, the sensitivity of the urine marker was as high as conventional markers such as AFP and PIVKA II. Synchronous examination of serum and urine markers showed a higher sensitivity than the single serum or urine marker for hepatocellular carcinoma. The sensitivity for non-advanced hepatocellular carcinoma was 50%, while that for advanced hepatocellular carcinoma was 83%. The urine marker may be useful to detect non-advanced hepatocellular carcinoma. The sensitivity for lung cancer was 83% and that for Stage I or II was 82%. Concerning uterine cervical tumor, the value of the urine marker increased with the grade of dysplasia. The sensitivity for ovarian carcinoma was 100%, while that for benign ovarian tumor was 0%. These findings suggest that urine diacetylspermine is a useful tumor marker in hepatocellular carcinoma, lung cancer and gynecologic malignancy as well as pancreatobiliary carcinoma. PMID- 15796047 TI - [Pitfalls in homogeneous assays for HDL-c and LDL-c in serum]. AB - Between 1994 and 2004, homogenous assays for HDL-C and LDL-C based on different determination principles were developed to replace complicated conventional precipitation methods. Nowadays, most laboratories employ homogenous assays. However, due to differences in principles and reactivity, measurements made by different assays do not necessarily match in some cases. HDL-C determinations may vary depending on duration and conditions of serum storage for the CDC-DCM and the homogenous assay methods, due to their different principles of determination. In patients with cholestasis, apoE-rich HDL, Lp-X and Lp-Y are occasionally observed. In these cases, the reactivity of homogenous assays for HDL-C varies markedly among the manufacturers. Furthermore, because the specific gravities of Lp-X and Lp-Y are comparable to that of LDL, these lipoproteins are grouped as LDL by ultracentrifugation, and this is a source of confusion in clinical settings. The American CDC assesses the accuracy of cholesterol assays by the BQ method, which measures the total of IDL, the narrowly-defined LDL, and Lp(a). However, of the various homogenous assays for LDL-C, reactivities to IDL and Lp (a) differ, and as a result, it is possible that type III hyperlipidemia characterized by increased IDL may be misdiagnosed. PMID- 15796048 TI - [PR of clinical testing]. AB - Regardless of its vital role of providing quality of patients care and optimizing overall healthcare costs, a clinical testing in Japan bears too strong cost containment pressures, which lead to too economically driven cost reduction initiatives. As a result, there is a risk of hampering infrastructure of appropriate medical services in Japan, including accuracy of tests, speed of tests, adoptation of new IVD technology, quality of information, adding value information, risk management, etc. In order to maintain a proper clinical testing in Japan, people/organizations associated with IVD and clinical testing should take more proactive PR actions to appeal the importance and value addition of clinical testing to patients, media, governments, hospital management, and other medical professionals. PMID- 15796049 TI - [New monitoring test for sleep apnea syndrome]. AB - We introduce change in pulse transit time (change in PTT: noninvasive intrathoracic pressure monitoring) as a new monitoring test of sleep apnea syndrome, which is being developed for clinical application. In addition, we report the kinetics of cardiogenic oscillation (CGO: noninvasive upper airway obstruction monitoring), which was published in this journal in 2003, and the response of the upper airway in clinical practice (under an endoscope). We also review diaphragmatic movement by abdominal echography (respiration generator monitoring). PMID- 15796050 TI - [Oxidative stress related diseases and biopyrrins]. AB - Reactive oxygen species (ROS) cause damage to the structure and function of tissues. Therefore tissues have systems that eliminate ROS. Bilirubin is one antioxidant that reacts with ROS to produce oxidative metabolites. Biopyrrins are one of the metabolites, the level of which in urine reflects oxidative stress. They are measured by non-competitive inhibition ELISA that employs anti-bilirubin antibody (24G7) and the results are corrected for the urinary concentration of cereatinine. Some reports suggested that psychological stress increased oxidative stress markers. Urinary biopyrrins were also elevated by speech stress, and the subjective stress score recorded by the speakers correlated with the level. The result suggests that bilirubin might eliminate ROS generated by psychological stress. From the beginning of the study of biopyrrins, their urinary level has been known to be increased by surgical stress. Furthermore, it was significantly higher in a major operation patient group than in a minor one, and correlated with operation duration. Sepsis increased the level in surgical patients. Ischemia-reperfusion elevates ROS and, as a result, biopyrrin production. An increase in urinary biopyrrins was observed in a coronary spastic angina group after a spasm provocation test, and the level in myocardial infarction patients with NYHA (New York Heart Association) classification became higher. Correlation between urinary biopyrrins and plasma B-type natriuretic peptide (BNP) was also reported. Research that determines the structures of biopyrrins and their clinical application are in progress. PMID- 15796053 TI - [Desirable role of the clinical laboratory in hospital infection control]. AB - Prevention of hospital-acquired infection is the most important strategy to control infection in terms of the well being of patients and the medical economy. Infection control in hospitals is carried out by the action against hospital based infection, for which clinical laboratory functions, such as surveillance and outbreak investigation, should be primary responsible. Therefore, it is essential for clinicians to have clinical laboratory rapidly detect pathogenic organisms and provide new and update information on appropriate antibiotics and clinical isolates. It is also desirable for clinical laboratory to collaborate with infection control team (ICT) and link nurse, and provide them with useful information on on-going infection. PMID- 15796052 TI - [Cooperation of the clinical laboratory and infection control department in prevention of hospital infections]. AB - Recently, environments that surround hospitals are changing every moment, and hospital infections are no exceptions. As medical technology advances and population of the elderly grow larger steadily, so does the number of immune compromised hosts. Many patients come to hospitals infected, and in such situations prevention of hospital infections has great amount of importance. Cooperation of microbiological laboratory and ICT is indispensable, accumulation and analysis of the data of resistant bacteria isolated from various medical materials are the biggest task of microbiological lab. Furthermore, nutrition support based on the values of clinical examinations is set as the ultimate goal and NST's cooperation plays a big part in supporting the control of hospital infections. PMID- 15796054 TI - [Molecular mechanism in life style diseases]. PMID- 15796051 TI - [Central clinical laboratory and infection control team: an essential partnership controlling health care-associated infections]. AB - Infection Control Committee (ICC) of Akita University Hospital was established in 1980's, when methicillin-resistant Staphylococcus aureus (MRSA) spread in teaching hospitals in Japan. After 20 years from the establishment of ICC, we needed to shake up some of the outdated infection control systems. Infection Control Team (ICT) was established in 2002 to reinforce ICC. ICT was consisted of five infection control doctors (ICD), two infection control nurses (ICN), two medical technicians in bacteriological division of clinical laboratory, one pharmacist and three administrative officials in the hospital. Monitoring of multiple drug-resistant pathogens, antibiotic use, surgical site infections and bloodstream infections are mandatory. Personnel training and educational activity are also required. After the establishment of ICT, inappropriate use of antibiotics and prevalence of MRSA were dramatically decreased. The hospital saved more than 30,000,000 yen annually. However, with a great regret, we experienced an outbreak of MRSA in a department of our hospital in 2003. MRSA infection was judged as a cause of death in three patients in the outbreak. MRSA was thought to transmit via medical personnel since pulsed-field gel electrophoresis revealed common genotype in 11 out of 15 patients studied. Prevention of healthcare-associated infections is a crucial in the management of hospitals. In this paper, we verified the efforts to control the outbreak and analyzed factors interfering infection control activities. A crucial role of a clinical laboratory in controlling healthcare associated infections was also discussed. PMID- 15796055 TI - [Diagnosis of and therapy for peptic ulcer]. PMID- 15796057 TI - [Amyloidosis in patients undergoing long-term hemodialysis: elucidation of etiological mechanism and therapeutic stratigy]. PMID- 15796056 TI - [Therapies for intractable respiratory infection due to Pseudomonas aeruginosa]. PMID- 15796058 TI - [Recent progress and problems in percutaneous coronary intervention]. PMID- 15796059 TI - [Studies on pathogenesis of Parkinson's disease]. PMID- 15796060 TI - [Recent progress in study and therapy for COPD]. PMID- 15796061 TI - [Molecular target therapy for malignant tumors]. PMID- 15796063 TI - [Generalization and specialization in the field of internal medicine (discussion)]. PMID- 15796062 TI - [Life style and digestive system diseases]. PMID- 15796064 TI - [Neuroadaptive mechanisms form development of psychological dependence on volatile organic solvents]. AB - Abuse of volatile organic solvents among youth remains a major social problem. Organic solvents are cheap and relatively easy to obtain, so they carry the risk of becoming a so-called "gateway drug" for users. Most research regarding organic solvents has until now focused on their neurotoxicity, specifically examining the mechanism of neuron death in terms of the involvement of substances such as nerve growth factor. However, systems to assess psychological dependence on volatile organic solvents that take into account the mechanism involved in the development of this dependence have not been established due to the difficulty of creating animal models. The conditioned place preference procedure, which can easily assess whether psychological dependence has been formed, has been phased in in recent years, and dependence assessment systems have been established for drug inhalation. There have also been new research developments regarding dependence on volatile organic solvents. The importance of mesolimbic dopamine neurons has been indicated in the expression of CNS stimulant action and the development of psychological dependence on drugs such as stimulants, cocaine, and heroin, which are typical abused drugs. It has recently become apparent that the increase in dopamine release in the nucleus accumbens accompanying activation of mesolimbic dopamine neurons, as has conventionally been proposed, is important to the expression of CNS stimulant action and the formation of psychological dependence in response to inhalation of toluene, a volatile organic solvent. Furthermore, research with regard to organic solvents' site of action is also proceeding based on studies using molecular biological techniques. Research regarding toluene is progressing, and the importance of receptors that gate ion channels such as N methyl-D-aspartate (NMDA) and y-aminobutyric acid (GABA)A receptors as candidates for toluene's site of action has been indicated. Clarification of organic solvents' mechanism for the development of psychological dependence is expected to progress, thanks to analysis focusing on such new sites of action. PMID- 15796065 TI - [Regulation of TH activity by GAD immunoreactivity in rat peri-LC after chronic variable stress]. AB - Locus coeruleus (LC) is the major component of noradrenergic neurons in the brain. The corticotropin-releasing hormone (CRH) and norepinephrine (NE) are suggested to play a role in modulating the central stress response. In a previous study we observed a decrease of the basal level of tyrosine hydroxylase (TH) immunoreactivity (-ir) in the LC of rats treated with chronic variable stress (CVS) for 14 days. Furthermore a novel stressor produced an enhanced response of the TH-ir after CVS. In the present study we examined the effect of CVS on the glutamic acid decarboxylase (GAD)-ir activity of periaqueductal gray (PAG), prepositus hypoglossi (PrH) and peri-LC. The GAD-ir was significantly increased in PrH and peri-LC after CVS. The footshock-induced reactivity in the GAD-ir was decreased in both regions after CVS. Moreover, we investigated the influence of the CRH receptor antagonist, alphah-CRH (i.c.v.) on the CVS-induced activation of the TH-ir in the LC. The alphah-CRH i.c.v. diminished the enhanced-TH reactivity by novel stressor after CVS. Our results suggest that the GABA activity in peri LC and PrH might regulate the LC-TH response, and also the CRH input from central nucleus of amygdala (CeA) and/or the bed nucleus of stria terminalis (BNST) might regulate the TH reactivity. PMID- 15796066 TI - [Effect of chronic variable stress on limbic corticotrapin-releasing hormone receptor]. AB - The Corticotropin-releasing hormone (CRH) is a key mediator in the stress response. Two CRH-receptor subtypes have been identified in the brain, CRH receptor 1 (CRH-R1) and CRH-receptor 2, and stress responses are mediated by the CRH-R1. In this study we have examined the effect of chronic variable stress (CVS) on the CRH-R1 immunoreactivity in the hypothalamic and limbic brain regions of the rat. For CVS we selected the six stressors described in previous reports. Male Wistar rats were randomly exposed to one stressor per a day during 13 days and finally exposed to a novel stressor (footshock). There was a significant increase of the CRH-R1 immunoreactivity 24 hours after CVS in the bed nucleus of the stria terminalis (BNST) and basolateral amygdala (BLA), whereas a single footshock had no influence on the immunoreactivity in any of the regions. A novel stressor after CVS suppressed within 24 hours the CVS-induced increase in the CRH R1 immuoreactivity in the BNST and BLA. Our results suggest that an up-regulation of the limbic CRH-R1 may contribute to stress sensitization (vulnerability) such as an anxiety, arousal and hypersympathetic autonomic symptoms under stressful situations. PMID- 15796067 TI - [Mechanisms of antidepressants and serotonin (5-HT)-induced glial cell line derived neurotrophic factor (GDNF) releases in rat C6 gliobrastoma cells]. AB - Recent studies show that neuronal and glial plasticity are important for the therapeutic action of antidepressants. Here, we demonstrated that amitriptyline, a tricyclic antidepressant, significantly increased GDNF mRNA and GDNF release in C6 cells. Furthermore, different classes of antidepressants increased GDNF release, but non-antidepressant psychotropic drugs did not. The amitriptyline induced GDNF release was completely inhibited by U0126, a mitogen-activated protein kinase (MAPK)-extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor, but was not inhibited by H-89, a protein kinase A inhibitor or calphostin C, a protein kinase C inhibitor. These results suggest that the amitriptyline-induced GDNF release may be regulated through a MEK/MAPK pathway. Next, we examined the effects of monoamines on GDNF release, because antidepressants are known to increase monoamines. 5-HT increased GDNF mRNA and GDNF release, but noradrenaline and dopamine did not. The 5-HT-induced GDNF release was partially, but significantly, blocked by ketanserin, a 5-HT2A receptor antagonist. The 5-HT-induced GDNF release was completely inhibited by U0126, but was not inhibited by H-89 or calphostin C. These results suggest that the 5-HT-induced GDNF release was mediated through a MEK/MAPK pathway and, at least, 5-HT2A receptors. GDNF, as well as other neurotrophic factors, may contribute to explain the therapeutic action of antidepressants and suggest a novel strategy of pharmacological intervention. PMID- 15796068 TI - [Pain and Bradykinin Receptors--sensory transduction mechanism in the nociceptor terminals and expression change of bradykinin receptors in inflamed condition]. AB - Bradykinin (BK), an endogenous algesic and sensitizing substance, excited nociceptors and sensitized their heat responses. These effects were mediated by B2 receptors (B2Rs) in normal condition, and B1 receptors were additionally recruited in inflammation. B2Rs were coupled with Gq/11 and their activation resulted in diacylglycerol and inositol triphosphate release. Diacylglycerol activated protein kinase (PK) Cepsilon in sensory neurons. To clarify what channel was modulated by PKC to depolarize nociceptor terminals, we examined the heat activation threshold (Tt) of heat-sensitive capsaicin receptor (TRPV1). Tt was lowered down to 31 degrees C by BK in concentration dependent manner through activation of PKCepsilon in cells heterologously expressing TRPV1 and B2Rs. Thus both excitation and sensitization to heat could be explained by one mechanism, lowering Tt of TRPV1. The same was observed in capsaicin-sensitive primary sensory neurons. However, TRPV1 knockout mice showed almost no change in BK induced nociceptive behavior and nociceptor excitation, although BK-induced heat hyperalgesia completely disappeared, suggesting that TRPV1 was not the sole channel that was modulated by BK to depolarize nociceptor terminals. In addition nociceptor sensitivity to BK was augmented in inflamed animals, with B2R mRNA and protein upregulated. The mechanism for prostaglandin-induced augmentation of BK response is left open for future study. PMID- 15796069 TI - [Study of Parkinson's disease-causing mechanism and development of anti Parkinson's disease drugs using endogenous substances in brain]. AB - There have been many reports showing 1,2,3,4-tetrahydroisoquinoline(TIQ) derivatives may be related to the onset of Parkinson's disease. 1-Benzyl-1,2,3,4 tetrahydroisoquinoline(1BnTIQ) was detected as a novel endogenous amine in mouse brain and parkinsonian CSF. The level of 1BnTIQ was very high in CSF of some parkinsonian patients compared with that of controls with other neurological diseases. Repeated administration of 1BnTIQ induced behavior abnormalities. We suggest that 1BnTIQ may be related to the idiopathic Parkinson's disease. On the other hand, 1-methyl TIQ (1MeTIQ), an endogenous amine in the brain, prevented 1BnTIQ-induced parkinsonism in mice. So 1MeTIQ is a candidate for anti parkinsonian drugs. PMID- 15796070 TI - [Plasticity of central monoaminergic systems during aging]. AB - The locus coeruleus (LC), located within the caudal pontine central gray, is composed of noradrenaline-containing neurons. The axons of these neurons form extensive collateral branches that project widely to many brain sites. The function of the LC is still unclear at present, however, LC neurons are known to exhibit marked axonal regeneration and sprouting in response to brain damage. We investigated the age-related changes in noradrenergic innervations of the frontal cortex, using in vivo electrophysiological techniques and immunohistochemistry. While noradrenergic innervations gradually decreased with age in the frontal cortex, a high degree of sprouting occurred in the LC axon terminals in middle age. Neither the electrophysiological properties of LC neurons nor NA levels in the frontal cortex changed with age. These findings suggested that the LC neurons preserve a strong capacity to remodel their axon terminals even in the aging brain. Exogenous brain-derived neurotrophic factor (BDNF) infusion caused a marked increase in the density of noradrenergic axon in the aged brain, but no trophic action of BDNF was observed in the young or middle-aged brain. The result suggests that BDNF is necessary for the maintenance of noradrenergic innervations in the aged brain. PMID- 15796071 TI - [Differentiation regulation of neural stem cell lines]. AB - We established clonal cell lines from fetal brains and a cerebellum of p53 deficient mice. In this paper, stem cell lines are selectively described. FBD 103a line generates neurons, astrocytes and oligodendrocytes. 2Y-6fl line generates neurons and astrocytes. 2Y-3t line generates neurons and oligodendrocytes. FBD-104 line generates astrocytes in conventional culture, but formation of spheres in suspension culture gives the line the capability to produce neurons in addition to astrocytes. Analyses of these lines were done mainly by limited dilution cloning, immunostaining and Western blotting. Some factors in culture medium and kidney were shown to regulate differentiation of the stem cell lines. PMID- 15796072 TI - Survey highlights infection control issues facing nurses. PMID- 15796073 TI - 'Media is wrong to blame nurses'. PMID- 15796074 TI - NT campaign will support nurses in the fight against MRSA infection. PMID- 15796075 TI - A voice from mental health. PMID- 15796076 TI - Keep it clean, back nurses to fight infection. PMID- 15796077 TI - How I coped with the tsunami disaster. PMID- 15796078 TI - Nurses' role in promoting and supporting smoking cessation. AB - Smoking is one of the biggest threats to public health, costing more than 120,000 lives per year in the UK alone (Peto et al, 2004). Helping smokers give up is one of the most effective health interventions and nurses are in a unique position to fill that role. PMID- 15796079 TI - Pleurisy. PMID- 15796080 TI - Medroxyprogesterone. PMID- 15796082 TI - Change from an office-based to a walk-around handover system. AB - This project stemmed from a desire to review the interdisciplinary handover system in an over-65s rehabilitation ward. The idea was to involve staff, including all members of the interdisciplinary team, in the decision-making process, and to canvass patients' opinions about the walk-around report. PMID- 15796081 TI - Measuring PEFR. PMID- 15796083 TI - Understanding the symptoms of irritable bowel syndrome. AB - This article discusses the symptoms, treatments and nursing management of patients who have irritable bowel syndrome. It has been recognised as the most common outpatient disorder in the U.K. Various terms have been used to describe the syndrome, including irritated colon, irritable colon syndrome, mucous colitis or spastic colon. PMID- 15796084 TI - Appreciating the benefits of effective clinical supervision. PMID- 15796085 TI - Helping your team to work effectively. PMID- 15796086 TI - Children are not just miniature adults. PMID- 15796087 TI - Update on the pathogenesis of osteolysis in multiple myeloma patients. AB - Multiple myeloma (MM) is a plasma cell malignancy characterized by the high capacity to induce osteolytic bone lesions that mainly result from an increased bone resorption related to the stimulation of osteoclast recruitment and activity. Although it is known that myeloma cells induce osteoclastic bone resorption, the biological mechanisms involved in the pathophysiology of MM induced bone resorption have been unclear for several years. Recently, new data seem to elucidate which mechanism is critically involved in the activation of osteoclastic cells in MM. The critical osteoclastogenetic factor RANKL and its soluble antagonist osteoprotegerin (OPG) are the major candidates in the pathophysiology of MM bone disease. Human MM cells induce an imbalance in the RANKL/OPG ratio in the bone marrow environment that triggers the osteoclast formation and activation leading to bone destruction. The role or RANKL/OPG system and other osteoclast stimulating factors in the pathophysiology of MM bone disease are summarized in this update. PMID- 15796088 TI - Adherence to physical activity in young people with type 1 diabetes. AB - Regular physical activity plays a key role in the management of children and adolescents with Type 1 diabetes mellitus but it is not considered as a treatment for diabetes. Aim of this study was to investigate time spent exercising, adherence to the programme for a safe exercise and ability of young people with diabetes to take appropriate measures to reduce potential risks. Ninety one Type 1 diabetes mellitus young people (aged from 10 to 18 years, duration of diabetes longer than 6 months) without associated chronic diseases were randomly enrolled in the study. Age, sex, weight, height, BMI, duration of disease, mean HbA1c value over preceding 6 months have been collected. The time weekly spent for physical activity, the type of exercise usually performed, the measures taken to reduce exercise risks have been collected by a structured questionnaire. BMI was 21.6+/-3.05 in the boys and 21.3+/-3.63 in the girls. All patients spent exercising 438+/-221 minutes/week. Boys exercised 71 minutes longer than girls in competitive sports. Children exercising less than 60 minutes weekly showed a mean HbA1c level (8,9+/-05%) higher than that found in children exercising 120-360 minutes (8,3+/-0.4 %; p=0.002) or 360-480 minutes (8,0+/-0.6 %; p< 0.01) weekly. Children attending a competitive sport (at least 360 min per week) had a better glycemic control (HbA1c=7,39+/-0.6 %; p=0.03) than other active peers. Fifty percent of patients reported to monitor blood glucose levels during exercise; 32 % changed insulin dose according to blood glucose levels; 60 % usually added carbohydrate-based foods before (35%), during (15%) or after (10%) exercise. Hypoglycemic episodes (37.7%) were reported more frequently than hyperglycemic ones (p=0.024), but only twelve percent of them were symptomatic and appeared 30 minutes to 2 hours after the end of exercise. These results must encourage health care professionals to review regularly the ability of their patients in managing physical activity and to check their adherence to the program for a safe exercise. PMID- 15796089 TI - Video-assisted thoracoscopy in thoracic injury: early or delayed indication? AB - The aim of this study was to evaluate the diagnostic role and therapeutic effectiveness of videothoracoscopy in chest trauma. Between 1st January 1993 and 30th September 2003, 112 traumatized patients underwent a videothoracoscopy. The origin of trauma was different: 60 road accidents, 42 accidental falls, 7 knife wounds and 3 firearm wounds. Seventy-six patients presented hemothorax, 21 hemopneumothorax, 10 suspected diaphragmatic injury and 5 pericardium effusion. All patients were hemodynamically stable and conscious. In suspected diaphragmatic rupture patients, videothoracoscopy confirmed the presence of 4 lesions with diameter from 7 to 10cm. The etiopathogenetic causes in the other clinical patterns were: 20 lung lacerations, 17 apical adhesion lacerations, 11 diaphragmatic lesions, 16 wall bleedings and 38 vessel lesions. Ninety patients (80.3%) were treated with video assisted thoracic surgery. The remaining procedures were : 17 drainage tube insertions, 4 thoracotomies and 1 laparotomy. Videothoracoscopy made the use of the primary intention drainage tube obsolete in stable traumatized patients with hemothorax or hemopneumothorax. It is a safe technique that allows the diagnostic and surgical management of the lesions. PMID- 15796090 TI - The DIAB.&TE.S Project: how patients perceive diabetes and diabetes therapy. AB - Assessment of quality of life (QoL) in diabetic patients is increasingly retained as a crucial parameter to take into account before concluding on the efficacy of new therapies. Indeed, the trend is to adopt treatments providing a genuine clinical benefit associated with a positive impact on QoL. In this context, DIAB.&TE.S Project aimed at analyzing patient's perception of psychological well being, quality of treatment and their correlation with glyco-metabolic values control. A total of 1918 diabetic patients, mostly from southern Italy, were enrolled in this observational study. DIAB.&TE.S included a wide range of patients taking insulin monotherapy, OHA (oral hypoglycemic agents) monotherapy or combined therapy (insulin + OHA). Internationally recognized instruments such as the Well-Being Questionnaire (WBQ) and Diabetes Treatment Satisfaction Questionnaire (DTSQ) were used to assess general well-being and diabetes treatment satisfaction. Combined therapy was not highly efficient in achieving both glycemic and quality of life goals when compared to insulin or OHA monotherapies. Oral monotherapy produced better results if compared to insulin treatment, as demonstrated by DTSQand WBQ scores. However, among the insulin treated subjects, higher scores were reached in patients using advanced devices to administrate insulin, Finally, a correlation between questionnaires outcomes and metabolic control in patients could be established. Our findings support the idea that in diabetes therapy, integration of clinical and psychosocial cares could be helpful in both identifying subjects that need a treatment re examination and improving patient's quality of life. In addition, this survey by its magnitude provides an overview of diabetes management in Italy and contributes to set up an accurate profile of diabetic people in this part of Europe. PMID- 15796092 TI - Eating behavior and body image perception: an epidemiological study on Italian adolescents. AB - In order to identify some of the psychological-environmental factors behind an unbalanced diet or incorrect eating habits which may be connected to the onset of eating-behavior disorders. A sample of 4550 teenagers (2221 males and 2329 females) aged between 12,3 and 18,9 years (mean age = 14.49+/-3.8), was consecutively recruited taken from several secondary schools and high schools. The Pisa Survey for Eating Disorders (PSQ), a questionnaire aimed at evaluating both eating habits and self-perception of body image, was administered. Answers given to some items have highlighted a recurrence of cognitive distortions, an excessively rigid self-perception of body image as well as frequent, often daily, dysfunctional eating habits. The PSED questionnaire has proved itself to be a useful and easy-to-use tool. It helps to reveal maladjusted behavior and to evaluate probable risk factors connected with the development of eating and emotional disorders. PMID- 15796091 TI - Strategies for optimizing compliance of paediatric patients for seasonal antibacterial vaccination with sublingually administered Polyvalent Mechanical Bacterial Lysates (PMBL). AB - The objective of this pilot study was to evaluate efficacy, tolerance and compliance of paediatric patients vis-a-vis a cycle of PMBL treatment (a sublingual tablet taken for ten consecutive days over three consecutive months). The study enrolled 89 children (65 randomised to the treated group and 24 to the control group). The study protocol included an enrolment check-up (TO) and follow ups at two months (T1), three months (T2) and nine months (T3) following the end of treatment, during which episodes of RRI were recorded; the main blood chemistry, immunology and phlogosis parameters were measured, together with hepatic, renal and bone marrow toxicity indexes. The administration of PMBL led to a significant decrease in RRI in the treated group, not only among the same children in relation to the previous winter, but also in comparison with untreated children during the same winter (mean number of infective episodes per patient 7.84 vs. 4.78, p<0.05, in the first case; 6.78 vs. 4.78, p<0.05, in the second case). White blood cell count showed a drop in the treated group as opposed to an increase in the untreated group, but there were no statistically significant differences in the intergroup analysis or in the intragroup one. Phlogosis indexes (PCR and plasma mucoprotein) in the treated group fell following treatment with PMBL, and this is statistically significant not only in the intragroup analysis but also the intergroup one. Mean values of B-lymphocytes in the treated group seemed to increase significantly following treatment, which was not the case in the untreated group. The variations in all the blood chemistry indexes for toxicity were far from significant and they remained within the norm, without significant clinical manifestations of side-effects of drug intolerance. As to evaluation of patient compliance, use of the device we describe enabled acceptable compliance with treatment even in the youngest children, similar to the compliance observed among appropriately motivated older children. PMID- 15796093 TI - Post-traumatic descending aorta intramural haematoma fortuitously witnessed during a magnetic resonance examination of the spine. AB - Most patients with traumatic aortic lesions of the thoracic aorta die at the accident site but 15% arrive at the hospital with vital signs. Digital subtraction angiography is considered the gold standard for the diagnosis of traumatic lesions of the thoracic aorta. Other non-invasive diagnostic techniques, such magnetic resonance and spiral computed tomography, proved to be reliable alternatives. Sometimes the diagnosis can be delayed by the relative lack of significant symptoms as well as the presence of unusual clinical findings. A case of post-traumatic intramural haematoma of the descending aorta revealed during a magnetic resonance of the spine is described. The final diagnosis was obtained with spiral computed tomography. PMID- 15796094 TI - Totally laparoscopic esophago-gastrectomy. AB - The recent progress of surgical endoscopy has allowed to perform minimally invasive esophagectomy both by a combined laparoscopic/thoracoscopic and a totally laparoscopic transhiatal approach. Up to now, all these techniques imply a thoracic (or thoracoscopic) and/or cervical access to perform the esophagogastric anastomosis. In the presented case, a 44-year-old white male patient affected by adenocarcinoma of the cardia (Siewert type 2) underwent a distal esophagectomy entirely accomplished by laparoscopy (including the realization of a mechanical intrathoracic esophagogastrostomy), without the need of any thoracic (nor thoracoscopic) nor cervical access. Duration of surgery was 407 minutes. Postoperative course was uneventful and the patient was discharged on day 7. In selected cases, a totally laparoscopic esophago-gastrectomy without thoracic nor cervical access, may improve the patient's outcome PMID- 15796095 TI - [Protecting health]. PMID- 15796096 TI - Effects of impulse noise on transiently evoked otoacoustic emission in soldiers. AB - The aim of the study was to assess the effects of exposure to impulse noise on TEOAE, as compared to PTA. The study comprised 92 soldiers, subjected to impulse noise during military service. The control group consisted of secondary school students, not exposed to noise. Extended high frequency PTA, and TEOAE were recorded before and after one year of military service. The total level of noise and spectrum analysis were performed for all kinds of weapons, separately. The highest levels of noise for weapons were related to frequencies from 1.6-16 kHz. After military service significant deterioration of hearing was observed on average by 6 dB exclusively at the frequencies of 10 and 12 kHz. TEOAE reduction was registered predominantly at frequencies of 2, 3 and 4 kHz, with the greatest decrease at 2 kHz (p <0.02). The control group did not show any significant audiometric changes as well as TEOAE during the time of experiment. PMID- 15796097 TI - Effects of body position on transient evoked otoacoustic emissions: the clinical perspective. AB - The present study investigated body position effects on transient evoked otoacoustic emission (TEOAE) recordings of clinical significance. Sixty adults (30 males, 30 females) were assessed using the Otodynamics ILO88 Analyzer in three positions (sitting, supine, and side-lying). Results indicated significant positional effects on the TEOAE parameters of A-B difference, noise, whole wave reproducibility, and response levels. These differences included higher noise levels in supine and side-lying positions in comparison to the upright sitting position. Lower whole wave reproducibility measurements, and higher response amplitudes, in the side-lying position compared with supine and seated positions were also observed. No significant effects were evident for signal-to-noise ratio or band reproducibility. Given the lack of significant body position effects on these latter parameters and the infrequent clinical use of the other parameters in isolation, there was no evidence to suggest the future need for major review of current pass/fail criteria or of the standard test protocol. PMID- 15796098 TI - Otoacoustic emissions in the general adult population of Nord-Trondelag, Norway: III. Relationships with pure-tone hearing thresholds. AB - This study aims to describe the association between otoacoustic emissions (OAEs) and pure-tone hearing thresholds (PTTs) in an unscreened adult population (N =6415), to determine the efficiency by which TEOAEs and DPOAEs can identify ears with elevated PTTs, and to investigate whether a combination of DPOAE and TEOAE responses improves this performance. Associations were examined by linear regression analysis and ANOVA. Test performance was assessed by receiver operator characteristic (ROC) curves. The relation between OAEs and PTTs appeared curvilinear with a moderate degree of non-linearity. Combining DPOAEs and TEOAEs improved performance. Test performance depended on the cut-off thresholds defining elevated PTTs with optimal values between 25 and 45 dB HL, depending on frequency and type of OAE measure. The unique constitution of the present large sample, which reflects the general adult population, makes these results applicable to population-based studies and screening programs. PMID- 15796099 TI - Otoacoustic emissions and extended high-frequency hearing sensitivity in young adults. AB - The relationship between hearing sensitivity in the extended high-frequency region (8-16 kHz) and (1) the presence of synchronized spontaneous otoacoustic emissions (SSOAEs) and (2) the strength of click-evoked and distortion product otoacoustic emissions (CEOAEs and DPOAEs) was investigated in 104 ears of 57 young adults. The age range was confined to 16 to 19 years. All subjects had normal hearing in the conventional audiometric frequency range (0.5-8 kHz). Ears with detected SSOAEs had better hearing sensitivity in the extended high frequency region and also higher levels of CEOAEs and DPOAEs than ears with undetected SSOAEs. The results support the hypothesis that the presence of SSOAEs is indicative of an ear with highly normal cochlear function over a broad frequency range. PMID- 15796100 TI - Analysis of counted behaviors in a single-subject design: modeling of hearing-aid intervention in hearing-impaired patients with Alzheimer's disease. AB - Clinical procedures related to patients with Alzheimer's Disease (AD) largely fail to address the patient's hearing. Given the challenges of this population, unconventional indicators of treatment efficacy may be required. Palmer et al (1999) reported on caregiver-tracked behaviors as outcome measures for hearing aid intervention. Using these data, hearing aid use and subsequent behavior was modeled as a first-order dynamic system, characterized by responses following an exponential time course. The results of such modeling suggest predictable outcomes of hearing aid intervention, or at least useful parameters of quantification (e.g. time-constant and steady-state response), permitting critical assessment of effects of intervention on negative behaviors versus hearing aid use, comparisons among behaviors, and/or comparisons of hearing-aid use patterns and behavior counts among patients. Use in this and other difficult to-test populations warrant further study to evaluate clinical efficacy of the analysis described. PMID- 15796101 TI - Appearances are deceptive? Long-term cognitive and central auditory sequelae from closed head injury. AB - The purpose of the present study was to examine possible signs of long-term cognitive and/or central auditory sequelae seven to eleven years after a closed head injury (CHI) of sufficient severity to cause scull fracture and/or brain contusion. Another purpose was that this investigation should be carried out in a group of recovered trauma victims with, to the individual, no known or minimal sequelae. A computer-based set of five cognitive tests and three central auditory tests were used in a group of formerly brain-injured patients who considered themselves as well recovered. Most of the participants did not report any signs of cognitive or auditory impairment. Tests of working memory capacity, verbal information processing speed, phonological processing and verbal inference-making ability were used. Auditory brain response (ABR), distorted speech audiometry (interrupted speech), and phase audiometry were used to test central auditory function. The initial severity of brain damage, i.e. status when the patient arrived at the emergency ward, was estimated with Swedish Reaction Level Scale (RLS). Cognitive shortcomings after CHI were demonstrated in a high percentage (59%, 13/22) of the cases seven to eleven years after the injury. Central auditory processing disorders (APD) were also demonstrated in a fairly high percentage (58%, 11/19) of the subjects. None of the correlations between RLS and the results on cognitive and central auditory tests reached statistical significance. However, there was a correlation between cognitive performance and the results on the central auditory tests used in this investigation. Eighty percent (8/10) of those participants with pathologies on ABR and/or phase audiometry and/or IS also failed on one or more of the cognitive tasks, compared to 44% (4/9) among those with no signs of APD. It is possible, many years after CHI, to observe cognitive shortcomings and APD in a relatively high percentage of CHI cases that are subjectively considered to be fairly well recovered. The cognitive tasks used in the study have proved to be a sensitive method to discover cognitive impairments. Long-term cognitive sequelae and APD could not be predicted from RLS scores. PMID- 15796102 TI - Do patients with benign paroxysmal positional vertigo receive prompt treatment? Analysis of waiting times and human and financial costs associated with current practice. AB - This study retrospectively analysed how 20 patients with posterior canal benign paroxysmal positional vertigo (BPPV) were managed from primary care, to treatment in tertiary care. The average time from first referral to treatment was 93 weeks, with an average of 58 weeks within primary care and 40 weeks within hospital care. At least 85% of cases had classical symptoms of BPPV and could have been easily identified by Primary Care Physicians at first referral, had they been trained to recognise and diagnose the condition. It was concluded that patients could be treated more efficiently and at less cost if the condition was identified at first referral in primary care, and treated in either primary care or dedicated BPPV clinics receiving referrals from primary care. A dedicated clinic for BPPV is recommended, which will substantially reduce waiting time for treatment and save primary care and hospitals time and money by avoiding unnecessary appointments and medication. PMID- 15796103 TI - Effect of stimulation rate on speech perception in adult users of the Med-El CIS speech processing strategy. AB - The primary aim of the study was to determine the effect of altering channel stimulation rate on the performance of adult cochlear implant users. Six adult users of the Med-El CIS processing strategy underwent tests of categorical identification of synthetic speech, tests of sentence recognition and tests of consonant recognition in three listening conditions: high channel stimulation rate (ranging from 1500 to 2020 pps/ch), a medium rate (800 pps/ch) and a low rate (400 pps/ch). Number of channels was held constant across rate conditions. With the categorical identification task, performance varied by acoustic cue type but did not vary with rate. With the consonant recognition task performance varied by phonological feature, but there was also no significant effect of rate. However, two subjects showed markedly reduced sentence scores at lower rates. Results suggests that higher stimulation rates with the CIS strategy may be beneficial to speech perception in some cases. PMID- 15796104 TI - Real-world vehicle emissions: a summary of the 14th coordinating research council on-road Vehicle Emissions Workshop. AB - The Coordinating Research Council held its 14th Vehicle Emissions Workshop in March 2004, where results of the most recent on-road vehicle emissions research were presented. We summarize ongoing work from researchers who are engaged in improving our understanding of the contribution of mobile sources to ambient air quality and emission inventories. Participants in the workshop discussed efforts to improve mobile source emission models, light- and heavy-duty vehicle emissions measurements, on- and off-road emissions measurements, effects of fuels and lubricating oils on emissions, as well as topics for future research. PMID- 15796105 TI - Evaluating inspection and maintenance programs: a policy-making framework. AB - This article presents a new analysis approach to design and evaluate motor vehicle inspection and maintenance (I/M) programs. The new approach, called I/M Design, uses real-world data to provide two resources not previously available: (1) a transparent framework to quantitatively illustrate the range of emission reductions available from I/M, and (2) a sensitivity analysis tool to evaluate how key variables affect I/M performance. In addition, the approach satisfies a policy-making information need--how to convey, in a logical and straightforward manner, the expected benefits from I/M without relying on modeling tools inaccessible to those outside the air quality field. The material presented in this article illustrates the new approach by estimating hydrocarbon (HC) emission reduction benefits available from enhanced I/M in southern California's South Coast Air Basin. I/M-Design estimates that enhanced I/M results in a 14-28% reduction in light-duty motor vehicle HC exhaust emissions; this estimate compares well to other California I/M program evaluations. Even more importantly, I/M-Design sensitivity analyses illustrate how I/M programs that implement stringent failure thresholds, motivate pre-test repair work, and improve repair effectiveness can provide emission reductions that substantially exceed the performance of existing programs. PMID- 15796106 TI - Glazed tiles manufactured from incinerated sewage sludge ash and clay. AB - Sewage sludge incineration is applied extensively in highly populated cities as a final sludge treatment. In this study, incinerated ash was utilized as an additive to clay to manufacture glaze tiles. Four different amounts of ash (0, 15, 30, and 45%) were added, and five glaze concentrations (0.03, 0.06, 0.1, 0.15, and 0.2 g/cm2) were applied on the surface of biscuit tiles to study the effects of ash additive and glaze concentration on properties of fired samples. Sewage sludge was dehydrated and incinerated into ash at 800 degrees C. Subsequently, tile specimens were manufactured and fired at 800 degrees C to make biscuit tiles. Fritted glazes and iron oxide were used as the fundamental glaze and colorant, respectively. Finally, glaze was applied on the surface of biscuit tiles and then fired at 1050 degrees C to sinter them into glazed tile specimens. Tests were performed to analyze properties, including water absorption, firing shrinkage, weight loss on ignition, abrasion resistance, bending resistance, acid alkali resistance, and aging resistance on specimens of glaze tile. To further understand more about the microstructural behavior of glazed tile specimens, analysis of energy dispersive spectrometer, scanning electron microscopy, and X ray were carried out in this study. PMID- 15796107 TI - Extraction wells and biogas recovery modeling in sanitary landfills. AB - A general methodology is established that permits the characterization and evaluation of the optimum potential of biogas extraction at each vertical well in the sanitary landfill of Asturias, Spain. Twenty wells were chosen from a total of 225 for the study, and the maximum production flow of biogas, which is a result of the degradation of the municipal solid waste deposited within its area of influence, was determined for each well. It was found that this flow varied with time and is characteristic of each extraction well. The maximum extractable flow also was determined as a function of the composition of the biogas needed for its subsequent utilization. The biogas extraction yield in the wells under study varied between approximately 26 and 97%, with a mean recovery value of 82%. The low yields found in certain cases were generally caused by a sealing defect, which leads to excessive incorporation of air into the landfill gas through the surrounding soil or through the extraction shaft, and which make its subsequent utilization difficult. PMID- 15796108 TI - Filtration of fly ash using a fluidized-bed filter. AB - This study focuses on the control of particulates with a fluidized-bed filter in exhaust gas stream. The fluidized-bed filter classified in the granular bed filtration technology was employed to demonstrate the performance for removal of fly ash at indicated operating velocities, fixed bed heights, and bed temperatures; then the collecting mechanisms of particulates by fluidized-bed filter were studied. The size distribution of fly ash passing through the fluidized-bed filter was also analyzed. The results indicate that at higher operating velocities and fixed bed heights, the removal of fly ash is more efficient and inertial impaction is the main mechanism when the fluidized-bed is operated at room temperature (25 degrees C). While operating at higher temperatures (200 degrees C), efficiency of 93.2% to 99.4% can be achieved for submicron particles. It is supposed to be the diffusion mechanism that is responsible for collecting such small particles, and high temperature is a favorable condition because of diffusion. PMID- 15796109 TI - Determination of Schulze equation for chemical oxygen demand reduction using stabilized landfill refuse media. AB - Three columns each with 770 cm2 of surface area and 60-105 cm effective depth were set up for this study. These columns were filled with compacted, stabilized refuse. High-strength brewery wastewaters were uniformly trickled down the medium. Overall, 16 runs with various organic loadings were tested and the results demonstrated that the stabilized refuse had excellent capability in removing chemical oxygen demand (COD). The COD removal efficiency reached 95% at a depth of 60 cm at Q = 8 L/day for the initial COD of approximately 6000 mg/L and the efficiency increased to >99% at a depth of 90 cm (organic loading of 0.69 kg/m3/day). As would be expected, the filter performance is a function of flow rate, influent COD concentration, and bed depth. The Schulze equation is able to predict the COD removal performance well. The variations of pH, oxidation reduction potential, and volatile fatty acids indicated that the acidogenesis reaction occurred in the upper layers. PMID- 15796110 TI - Evaluation of trickle-bed air biofilter performance under periodic stressed operating conditions as a function of styrene loading. AB - Trickle-bed air biofilters (TBABs) are suitable for treating volatile organic compounds (VOCs) at a significantly high practical loading because of their controlled environmental conditions. The application of TBAB for treating styrene contaminated air under periodic backwashing and cyclical nonuse periods at a styrene loading of 0.64-3.17 kg chemical oxygen demand (COD)/m3 x day was the main focus of this study. Consistent long-term efficient performance of TBAB strongly depended on biomass control. A periodic in situ upflow with nutrient solution under media fluidization, that is, backwashing, was approached in this study. Two different nonuse periods were employed to simulate a shutdown for equipment repair or during weekends and holidays. The first is a starvation period without styrene loading, and the second is a stagnant period, which reflects no flow passing through the biofilter. For styrene loadings up to 1.9 kg COD/m3 x day, removal efficiencies consistently above 99% were achieved by conducting a coordinated biomass control strategy, that is, backwashing for 1 hr once per week. Under cyclical nonuse periods for styrene loadings up to 1.27 kg COD/m3 x day, stable long-term performance of the biofilter was maintained at more than 99% removal without employing backwashing. No substantial impact of nonuse periods on the biofilter performance was revealed. However, a coordinated biomass control by backwashing subsequently was unavoidable for attaining consistently high removal efficiency at a styrene loading of 3.17 kg COD/m3 x day. As styrene loading was increased, reacclimation of the biofilter to reach the 99% removal efficiency following backwashing or the nonuse periods was delayed. After the non-use periods, the response of the biofilter was a strong function of the biomass in the bed. No significant difference between the effects of the two different nonuse periods on TBAB performance was observed during the study period. PMID- 15796111 TI - Effects of ceiling-mounted HEPA-UV air filters on airborne bacteria concentrations in an indoor therapy pool building. AB - The purpose of this study was to assess the effectiveness of a new generation of high-volume, ceiling-mounted high-efficiency particulate air (HEPA)-ultraviolet (UV) air filters (HUVAFs) for their ability to remove or inactivate bacterial aerosol. In an environmentally controlled full-scale laboratory chamber (87 m3), and an indoor therapy pool building, the mitigation ability of air filters was assessed by comparing concentrations of total bacteria, culturable bacteria, and airborne endotoxin with and without the air filters operating under otherwise similar conditions. Controlled chamber tests with pure cultures of aerosolized Mycobacterium parafortuitum cells showed that the HUVAF unit tested provided an equivalent air-exchange rate of 11 hr(-1). Using this equivalent air-exchange rate as a design basis, three HUVAFs were installed in an indoor therapy pool building for bioaerosol mitigation, and their effectiveness was studied over a 2 year period. The HUVAFs reduced concentrations of culturable bacteria by 69 and 80% during monitoring periods executed in respective years. The HUVAFs reduced concentrations of total bacteria by 12 and 76% during the same monitoring period, respectively. Airborne endotoxin concentrations were not affected by the HUVAF operation. PMID- 15796112 TI - Emissions of polychlorinated dibenzo-p-dioxins and dibenzofurans from stack gases of electric arc furnaces and secondary aluminum smelters. AB - This study investigates the emissions of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) from four electric arc furnaces (EAFs) and eight secondary aluminum smelters (secondary ALSs) in Taiwan. The mean PCDD/F International-Toxicity Equivalents (I-TEQ) concentrations in the stack gases of these EAFs and secondary ALSs are 0.28 ng I-TEQ/Nm3 (relative standard deviation [RSD]= 100%) and 3.3 ng I-TEQ/Nm3 (RSD = 260%), respectively. The high RSDs, especially for those obtained from secondary ALSs, could be caused by the intrinsic differences in their involved feeding materials, furnace operating conditions, and air pollution control devices. The mean I-TEQ emission factor of PCDD/Fs for EAFs (1.8 microg I-TEQ/tonne-feedstock) is lower than that for secondary ALSs (37 microg I-TEQ/tonne-feedstock). This result might be because the involved furnace temperatures for secondary ALSs (650-750 degrees C) are lower than those for EAFs (1600-1700 degrees C), resulting in the deterioration of the combustion condition, leading to the formation of PCDD/Fs during the industrial process. This study found that the total PCDD/F emissions from EAFs (20 g I-TEQ/yr) and secondary ALSs (18 g I-TEQ/yr) are approximately 27, 53, and approximately 24, 49 times higher than those from municipal solid waste incinerators (MSWIs; 0.74 g I-TEQ/yr) and medical waste incinerators (MWIs; 0.37 g I-TEQ/yr), respectively; while those are 44 and 40% of total PCDD/F emission from sinter plants (45 g I-TEQ/ yr), respectively. Considering a more stringent emission limit has been applied to waste incinerators (0.1 ng I-TEQ/Nm3) in Taiwan lately, the results suggest that the control of the emissions from metallurgical processes has become the most important issue for reducing the total PCDD/F emission from industrial sectors to the ambient environment. PMID- 15796113 TI - Investigation of sources of atmospheric aerosol at a hot spot area in Dhaka, Bangladesh. AB - Samples of fine and coarse fractions of airborne particulate matter were collected at the Farm Gate area in Dhaka from July 2001 to March 2002. Dhaka is a hot spot area with very high pollutant concentrations because of the proximity of major roadways. The samples were collected using a "Gent" stacked filter unit in two fractions of 0- to 2.2-microm and 2.2- to 10-microm sizes. The samples were analyzed for elemental concentrations by particle-induced X-ray excitation (PIXE) and for black carbon by reflectivity methods, respectively. The data were analyzed by positive matrix factorization (PMF) to identify the possible sources of atmospheric aerosols in this area. Six sources were found for both the coarse and fine PM fractions. The data sets were also analyzed by an expanded model to explore additional sources. Seven and six factors were obtained for coarse and fine PM fractions, respectively, in these analyses. The identified sources are motor vehicle, soil dust, emissions from construction activities, sea salt, biomass burning/brick kiln, resuspended/fugitive Pb, and two-stroke engines. From the expanded modeling, approximately 50% of the total PM2.2 mass can be attributed to motor vehicles, including two-stroke engine vehicle in this hot spot in Dhaka, whereas the PMF modeling indicates that 45% of the total PM2.2 mass is from motor vehicles. The PMF2 and expanded models could resolve approximately 4% and 3% of the total PM2.2 mass as resuspended/fugitive Pb, respectively. Although, Pb has been eliminated from gasoline in Bangladesh since July 1999, there still may be substantial amounts of accumulated lead in the dust near roadways as well as fugitive Pb emissions from battery reclaimation and other industries. Soil dust is the largest component of the coarse particle fraction (PM2.2-10) accounting for approximately 71% of the total PM2.2-10 mass in the expanded model, whereas from the PMF modeling, the dust (undifferentiated) contribution is approximately 49%. PMID- 15796114 TI - Exhaust temperature profiles for application of passive diesel particulate filters to solid waste collection vehicles in California. AB - To reduce public exposure to diesel particulate matter (DPM), the California Air Resources Board has begun adoption of a series of rules to reduce these emissions from in-use heavy-duty vehicles. Passive diesel particulate filter (DPF) after treatment technologies are a cost-effective method to reduce DPM emissions and have been used on a variety of vehicles worldwide. Two passive DPFs were interim verified in California and approved federally for use in most 1994--2002 engine families for vehicles meeting min engine exhaust temperature requirements for successful filter regeneration. Some vehicles, however, may not be suited to passive DPFs because of lower engine exhaust temperatures. The purpose of this study was to determine the applicability of two types of passive DPFs to solid waste collection vehicles, the group of vehicles for which California recently mandated in-use DPM reductions. We selected 60 collection vehicles to represent the four main types of collection vehicle duty cycles--rolloffs, and front-end, rear, and side loaders--and collected second-by-second engine exhaust temperature readings for one week from each vehicle. As a group, the collection vehicles exhibited low engine exhaust temperatures, making the application of passive DPFs to these vehicles difficult. Only 35% of tested vehicles met the temperature requirements for one passive DPF, whereas 60% met the temperature requirements for the other. Engine exhaust temperatures varied by vehicle type. Side and front end loaders met the engine exhaust temperature requirements in the greatest number of cases with approximately 50-90% achieving the required regeneration temperatures. Only 8-25% of the rear loader and roll-off collection vehicles met the engine exhaust temperature requirements. Solid waste collection vehicles represent a diverse fleet with a variety of duty cycles. Low engine exhaust temperatures will need to be addressed for successful use of passive DPFs in this application. PMID- 15796115 TI - Quality of life of caregivers. AB - This article is a descriptive analysis of various features of the quality of life (QoL) of the caregivers in different pathological conditions. Definitions of the concept of QoL, caregiver goals and burdens, caregiver stress and coping with factors that could contribute to or impact the QoL in caregivers are discussed both through a review of literature and an analysis of empirical evidences. The QoL in caregivers of patients with Parkinson's disease is also discussed. An original Scale of QoL of Caregivers is described and compared with other existing measures. The concepts of patient-caregiver dyad in research and in interventions is stressed. PMID- 15796118 TI - Improvement in cognitive and language skills from preschool to adolescence in autism. AB - This paper reports on the developmental progression of a sample of 48 adolescents and young adults with autism who were previously assessed at preschool age and again in the mid-school period. In contrast to the earlier period when about one third of the children made dramatic gains, cognitive and language skills tended to remain stable or decline over this time span. The gain in mental and language age of the non-retarded adolescents with autism was less than half the change in their chronological age. The mentally retarded adolescents with autism showed some gain in mental age over time but this was far less than their change in chronological age, and they showed almost no gain in language age. Early childhood predictors of language skills in adolescence were functional play skills, responsiveness to others' bids for joint attention, and the frequency of requesting behaviors. PMID- 15796117 TI - Access and service use by children with autism spectrum disorders in Medicaid Managed Care. AB - Although Medicaid is the largest public payer of behavioral health services, information on access and utilization of services is lacking, and no data on the frequency of service use or types of services provided for children with autism spectrum disorders (ASDs) are available. As states move toward managed care approaches for their Medicaid program, services information is critical. Behavioral health service data for children with autism spectrum disorders were collected from a state Medicaid Managed Care (MMC) program and analyzed from fiscal years 1995 through 2000. Findings revealed that the number of children who received services over time increased significantly; however, the rate of service use was only one tenth of what should be expected based on prevalence rates. The mean number of service days provided per child decreased significantly, about 40%, and the most prevalent forms of treatment changed. Day treatment vanished and medication and case management increased disproportionately to the number of children served. Explanations and implications of the findings are presented as well as recommendations for future research. PMID- 15796119 TI - Diagnostic assessment of Asperger's disorder: a review of five third-party rating scales. AB - Five rating scales for screening and detection of Asperger's Disorder, three commercially available and two research instruments, are evaluated with reference to psychometric criteria outlined by Bracken in 1987 (Journal of Psychoeducational Assessment, 4, 313). Reliability and validity data reported in examiner's manuals or published reports are reviewed. The scales included in the review are the Asperger Syndrome Diagnostic Scale (ASDS), Autism Spectrum Screening Questionnaire (ASSQ), Childhood Asperger Syndrome Test (CAST), Gilliam Asperger's Disorder Scale (GADS), and Krug Asperger's Disorder Index (KADI). All published rating scales demonstrated significant weaknesses, particularly in the use of questionable normative samples. Among the published instruments, the KADI appears to be the most sound in terms of reliability and validity. The research instruments present incomplete psychometric data to date, but hold promise as clinical instruments. PMID- 15796120 TI - Psychometric properties of the Chinese version of the Psycho-educational Profile Revised (CPEP-R). AB - The purpose of this study was to examine the reliability and validity of the Chinese version of the Psycho-educational Profile-Revised (PEP-R). The Chinese PEP-R (CPEP-R) was administered to 63 preschool children with symptoms of autistic disorder recruited from special child-care centers in Hong Kong. Results showed that the scales of the CPEP-R were internally consistent, reliable across raters and temporally stable. Regarding the concurrent validity of the CPEP-R, the developmental score and developmental age assessed by the CPEP-R were significantly correlated with the Merrill-Palmer Scale of Mental Tests and the Hong Kong Based Adaptive Behavior Scale. The Behavioral Scale of the CPEP-R was also significantly related to the Childhood Autism Rating Scale. Besides replicating the findings in the Western context, the present study suggests that the psychometric properties of the PEP-R are stable across cultures and the related findings support the cross-cultural reliability of the tool. PMID- 15796116 TI - Neuropsychological functioning associated with high-altitude exposure. AB - This article focuses on neuropsychological functioning at moderate, high, and extreme altitude. This article summarizes the available literature on respiratory, circulatory, and brain determinants on adaptation to hypoxia that are hypothesized to be responsible for neuropsychological impairment due to altitude. Effects on sleep are also described. At central level, periventricular focal damages (leuko-araiosis) and cortical atrophy have been observed. Frontal lobe and middle temporal lobe alterations are also presumed. A review is provided regarding the effects on psychomotor performance, perception, learning, memory, language, cognitive flexibility, and metamemory. Increase of reaction time and latency of P300 are observed. Reduced thresholds of tact, smell, pain, and taste, together with somesthetic illusions and visual hallucinations have been reported. Impairment in codification and short-term memory are especially noticeable above 6,000 m. Alterations in accuracy and motor speed are identified at lower altitudes. Deficits in verbal fluency, language production, cognitive fluency, and metamemory are also detected. The moderating effects of personality variables over the above-mentioned processes are discussed. Finally, methodological flaws found in the literature are detailed and some applied proposals are suggested. PMID- 15796121 TI - The application of short forms of the Wechsler Intelligence scales in adults and children with high functioning autism. AB - We evaluated the predictive accuracy of short forms of the Wechsler intelligence scales for individuals with high functioning autism. Several short forms were derived from participants who had received the full procedure. Stepwise multiple regression analyses were performed to determine the strength of association between the subtests included in the short form and IQ scores based upon the full test. These analyses were performed for all participants, and also for autism participants with atypical subtest profiles. In all analyses the percentages of explained variance were typically in the .8-.9 range. It was concluded that short forms may be used with good predictive accuracy in individuals with high functioning autism, even when the subtest profile is atypical. PMID- 15796122 TI - The psychometric properties of the Vineland Adaptive Behavior Scales in children and adolescents with mental retardation. AB - The psychometric properties of the Vineland Adaptive Behavior Scales Survey Form were studied in a total population of children and adolescents with MR, and in the specific levels of functioning (n=826, age 4-18 years). The original division into (sub)domains, as assigned by the authors, was replicated in the total population and in the mild and moderate levels of functioning. In the severe and profound levels of functioning the structure was less well recognized. The reliability of the instrument proved to be good in the total population and the subgroups. The construct validity was high in all groups. The implications of these findings are discussed with respect to the usefulness of the Vineland for the population with MR. PMID- 15796123 TI - Weak central coherence and its relations to theory of mind and anxiety in autism. AB - Recent theory and research suggests that weak central coherence, a specific perceptual-cognitive style, underlies the central disturbance in autism. This study sought to provide a test of the weak central coherence hypothesis. In addition, this study explored the relations between the weak central coherence hypothesis, theory of mind skills, and social-emotional functioning in a group of high functioning children with autism. Results revealed equivocal support for the weak central coherence hypothesis, but found moderate correlations between verbal weak central coherence and theory of mind measures. No significant findings were observed between weak central coherence measures and social-emotional functioning. PMID- 15796124 TI - Three types of source monitoring by children with and without autism: the role of executive function. AB - Earlier investigations have found mixed evidence of source monitoring impairment in autism. The present study examined three types of source monitoring ability in children with autism and typically developing children. In three different conditions, participants were presented with word lists after which they were required to recall the source of the word for reality, external and internal source monitoring tasks. Group differences were found across all three conditions, with the comparison group outperforming the children with autism. The pattern of performance across the three conditions, however, was comparable for the two groups. Specifically, performance was higher on the reality monitoring task than either the external or internal source tasks. We suggest that the overall impairment found for the children with autism may be due to broader impairments in executive function. PMID- 15796125 TI - Goal directed locomotion and balance control in autistic children. AB - This article focuses on postural anticipation and multi-joint coordination during locomotion in healthy and autistic children. Three questions were addressed. (1) Are gait parameters modified in autistic children? (2) Is equilibrium control affected in autistic children? (3) Is locomotion adjusted to the experimenter imposed goal? Six healthy children and nine autistic children were instructed to walk to a location (a child-sized playhouse) inside the psychomotor room of the pedopsychiatric centre located approximately 5 m in front of them. A kinematic analysis of gait (ELITE system) indicates that, rather than gait parameters or balance control, the main components affected in autistic children during locomotion are the goal of the action, the orientation towards this goal and the definition of the trajectory due probably to an impairment of movement planning. PMID- 15796127 TI - A paternally inherited duplication in the Prader-Willi/Angelman syndrome critical region: a case and family study. AB - The Prader-Willi/Angelman Critical Region (PWACR; Chromosome 15q11-13) is of interest as a potential locus for genes conferring susceptibility to autism spectrum disorders (ASD). This report describes a female proband referred for evaluation of a possible ASD. Genetic analyses indicated that the proband, her father and one of her sisters, carried a paternally derived interstitial duplication involving 15q11-13. The proband showed evidence of ASD (PDD-NOS), borderline mental retardation, mild hypotonia and joint laxity. Her father and her sister were of normal intelligence and neither was thought to have an ASD, although speech/language difficulties and some autistic type behaviours were reported to have been present early in the development of the sister. This is one of the first reports of a child with a paternal duplication and an autism spectrum disorder. More research is required to determine whether paternally derived duplications that involve 15q11-13 are associated with developmental impairments. PMID- 15796129 TI - Asperger's disorder with co-morbid separation anxiety disorder: a case report. PMID- 15796126 TI - Specific genetic disorders and autism: clinical contribution towards their identification. AB - Autism is a heterogeneous disorder that can reveal a specific genetic disease. This paper describes several genetic diseases consistently associated with autism (fragile X, tuberous sclerosis, Angelman syndrome, duplication of 15q11-q13, Down syndrome, San Filippo syndrome, MECP2 related disorders, phenylketonuria, Smith Magenis syndrome, 22q13 deletion, adenylosuccinate lyase deficiency, Cohen syndrome, and Smith-Lemli-Opitz syndrome) and proposes a consensual and economic diagnostic strategy to help practitioners to identify them. A rigorous initial clinical screening is presented to avoid unnecessary laboratory and imaging studies. Regarding psychiatric nosography, the concept of "syndromal autism"- autism associated with other clinical signs should be promoted because it may help to distinguish patients who warrant a multidisciplinary approach and further investigation. PMID- 15796128 TI - Brief report: autistic traits in twins vs. non-twins--a preliminary study. AB - Previous studies have suggested that among affected sib pairs with autism there is an increase in the frequency of twins over what would be expected in comparison to the prevalence of twins in the general population. In this study we sought to determine whether sub-threshold autistic traits were more pronounced in twins than in non-twins. The Social Responsiveness Scale (SRS) was administered in an epidemiologic twin sample (n=802) and in a separate population-based sample of non-twins ascertained from a local school district (n=255). For males (but not females), the mean SRS score was significantly higher among twins than among non twins. As has been suggested for autism, twin status may incur increased liability to subthreshold autistic symptomatology, particularly in males. PMID- 15796130 TI - My 19-year-old son has autism and moderate mental retardation. What can I do to make him less passive? PMID- 15796131 TI - Extemporaneous quality. PMID- 15796132 TI - Variability in the data generated by laboratories measuring D-values of bacterial spores. PMID- 15796133 TI - Bacterial adhesion: considerations within a risk-based approach to cleaning validation. AB - Pharmaceutical manufacturing processes are vulnerable to varying degrees of microbial challenge (hazard) quantifiable as microbial ingress, and microbial retention risks affecting raw materials and inputs to the final product. Control over these risks is exacted by both purposefully designed and incidental (or fortuitous) properties of the manufacturing processes. Within the manufacturing environment, equipment cleaning and hold processes are uniquely prone to microbial challenge yet paradoxically demonstrate the greatest potential for mitigation of these risks. Cognition of those components and contributing factors associated with microbial challenge are necessary to facilitate scientifically sound risk assessments. In the context of equipment cleaning and hold processes, risk assessments are necessary to identify and contrive conditions, which are truly worst case for the validation of the control of microbial challenge. A number of components contribute to the risk of microbial retention, yet the phenomenon of microbial adhesion to surfaces remains one of the most ubiquitous and perplexing. The dual purpose of this review is to primarily precis and provide in a single reference those multi-factorial features and variables contributing to bacterial adhesion, and secondly to provide a guide for interpretation of those considerations for integration into a risk-based approach to cleaning validation. PMID- 15796134 TI - Design, development and qualification of a microbiological challenge facility to assess the effectiveness of BFS aseptic processing. AB - A programme of work has been initiated to further the understanding of the impact of the environment surrounding a Blow/Fill/Seal (BFS) machine upon the microbiological quality of processed product. A dedicated facility (Challenge Room) has been constructed and qualified to provide for the production and containment of dispersions of micro organisms in air of a room housing an operating BFS machine of a given style and configuration. The facility achieves homogeneous distribution of generated dispersions throughout the Challenge Room air, including that within and close to the critical area in which aseptic BFS operations occur. Generated microbial dispersions can be maintained in the room over extended time periods (up to 600 min) at a desired concentration within the range 10(1) to 10(7) cfu m(-3). They can also be produced employing different cell types, including bacterial endospores, Gram-positive and Gram-negative vegetative cells and yeast cells. Effective containment of dispersions is achieved while 'cards of product' (vials in sets) are conveyed from the Challenge Room to an adjacent Packing and Storage Area. Decontamination of the room and the housed BFS machine is accomplished through exposure to chlorine dioxide gas at a concentration of 1.0 mg dm(-3) for 120 min at room temperature (approximately 23 degrees C). PMID- 15796135 TI - Establishment of critical contamination risk locations ("hot spots") in environmental monitoring by means of three-dimensional airflow analysis and particulate evaluation. AB - A practical approach for the qualification of the surrounding environment of the critical area in aseptic processing has been developed. This method uses three dimensional air velocity measurements combined with airborne particle monitoring. The analysis of the results obtained using the methods described in this article are beneficial in the selection of sample sites and frequencies and in refining personnel procedures and materials flow in aseptic processing. We propose that this improved qualification method can be widely applicable for both existing and new aseptic processing areas. This paper shows the results of one case study utilizing this method. The particle distribution map of a Grade B environment based upon extensive analysis was found to correspond to room airflow, as visualized by air vector mapping. The actual annual environmental monitoring data, which include airborne particles and microbes, as well as other microbial monitoring data, are also presented with respect to their relationship to the airflow pattern. PMID- 15796136 TI - Evaluating the formulae for integrated lethality in ethylene oxide sterilization using six different endospore forming strains of bacteria, and comparisons of integrated lethality for ethylene oxide and steam systems. AB - Bacterial endospores from six different species of bacteria were exposed to a spectrum of ethylene oxide (EtO) sterilizing conditions. Temperature was varied from 40 to 60 degrees C and the ethylene oxide concentration was varied from 300 to 750 mg/L. Relative humidity was maintained at 60+/-10% RH. The fraction negative procedure was used to determine the D value for each of the test conditions. Bacterial species tested included Bacillus atrophaeus ATCC # 9372, Bacillus smithii ATCC # 51232, Bacillus subtilis "5230" ATCC # 35021, Bacillus subtilis, DSM # 4181, Bacillus pumilus ATCC # 27142, and Geobacillus stearothermophilus ATCC # 7953. All spore preparations were inoculated on filter paper strips packaged in blue, sterilizable glassine pouches. G. stearothermophilus was the least resistant organism tested. The most resistant organisms tested were B. atrophaeus and B. subtilis "5230". The B. subtilis "5230" strain was slightly more resistant than B. atrophaeus at conditions of 54C and EtO concentrations of 400, 600, and 750 mg/L, as well as at 60C/750mg/L EtO. The other species were between these extremes. This empirical data allowed the application of the recently published formula for converting D values from one set of conditions to another and evaluations of accuracy. The measured D values also allowed the determination of Z values based on temperature variations. These formulae, when applied to process temperatures independent of gas concentration, result in a Z value of approximately 32 degrees C that appears to be similar for all species tested. These data support the application of the previously published formulae 1-6 and allow the same approach to integrated lethality for ethylene oxide processes as is commonly applied to steam sterilization. A review of steam sterilization and related principles was conducted for comparison of integrated lethality for these two methods of sterilization. Errors associated with D values, Z values, extrapolation, and integrated lethality for both methods of sterilization are discussed. PMID- 15796137 TI - Peritoneal dialysis-related infections recommendations: 2005 update. PMID- 15796139 TI - Practice guidelines: do we really need them? PMID- 15796138 TI - Peritoneal catheters and exit-site practices toward optimum peritoneal access: a review of current developments. AB - OBJECTIVE: This review updates the 1998 International Society for Peritoneal Dialysis (ISPD) recommendations for peritoneal dialysis catheters and exit-site practices (Gokal R, et al. Peritoneal catheters and exit-site practices toward optimum peritonealaccess: 1998 update. Perit Dial Int 1998; 18:11-33.) DESIGN: DATA SOURCES: The Ovid and PubMed search engines were used to review the Medline databases of January 1980 through June 2003. Searches were restricted to human data; primary key word searches included dialysis, peritoneal dialysis, and continuous ambulatory peritoneal dialysis cross referenced with access, catheter, dialysis catheter, peritoneal dialysis catheter, and Tenckhoff catheter. Related searches were provided via the PubMed related articles link. STUDY SELECTION: Reports were selected if they provided identifiable information on catheter design, catheter placement technique, and survival or placement complications. Reports without such data were excluded from review. Each study was then categorized by its characteristics: single-center or multicenter; retrospective or prospective; controlled trial, with or without random patient assignment; or review article. MAIN RESULTS: There are few randomized controlled evaluations testing how catheter design and/or placement influence long-term survival and function, and these are typically conducted at a single center. The majority of reports represent retrospective single-center experiences, and these are supplemented by occasional multicenter data registries. CONCLUSIONS: There is substantial variability in catheter outcomes between centers, and this variability is more closely correlated with operator and center characteristics than with catheter design. Some catheter designs appear to impact long-term catheter success, and, in some cases, specific patient characteristics and dialysis formats combine with specific catheter designs to influence catheter survival. Most reporters prefer two-cuff designs and placement of the deep cuff at an intramuscular location. Intramuscular cuff placement results in fewer pericatheter leaks and hernias, but makes catheter removal more difficult. High risk patients (those with previous pelvic surgery) benefit from visual inspection of the peritoneum during catheter placement, and in randomized controlled trials, catheters with pre-shaped arcuate subcutaneous segments ("swan neck" designs) reduce the risk of early drainage failure via "migration." PMID- 15796140 TI - Peritonitis and automated peritoneal dialysis: a therapeutic conundrum? PMID- 15796141 TI - Sterile peritonitis in the peritoneal dialysis patient. AB - Peritonitis is a serious and common problem in the peritoneal dialysis (PD) population. Abdominal pain, fever, and cloudy PD fluid usually heralds the onset of infective peritonitis. However, in up to 20% of cases, no organism is identified. In these situations, diagnosis can be made only by excluding a microbiological cause and performing a cytological examination of the PD fluid to determine the cellular or noncellular constituents. This review examines the differential diagnosis of sterile peritonitis and uses cytological examination to facilitate the appropriate diagnosis. PMID- 15796143 TI - Oral nutritional supplementation in peritoneal dialysis patients--does it work? PMID- 15796142 TI - Therapeutic dilemmas in the management of peritonitis. PMID- 15796144 TI - Icodextrin--what about the fractions? PMID- 15796145 TI - Oral protein-energy supplements in peritoneal dialysis: a multicenter study. AB - BACKGROUND: Protein-energy malnutrition is prevalent in peritoneal dialysis (PD) patients and is associated with increased morbidity and mortality. OBJECTIVE: To evaluate the impact of prophylactic treatment with an oral protein-energy supplement (Protenplus; Fresenius AG, Bad Homburg, Germany) on nutritional parameters in patients starting PD. DESIGN: Prospective, multicenter, randomized study of group A patients (Protenplus, n = 35) and group B (controls, n = 30), with evaluations at baseline and at 6 and 12 months. STATISTICAL METHODS: Efficacy of factors by linear mixed model analysis for repeated measurements, chi square, t-test, and Mann-Whitney test. OUTCOME PARAMETERS: Patient compliance, serum albumin, and other nutritional parameters. RESULTS: No significant differences were found at baseline evaluation. During follow-up, a significant number of group A patients abandoned intake of the supplement due to non compliance (n = 7) or side effects (n = 8) (chi2 p < 0.01). Patients with lower residual renal function were less likely to comply. The mixed model in the "intention to treat" analysis showed a significant increase related to supplement intake only in total lymphocyte count in group A. The "as treated" analysis of the 29 patients who fulfilled the study (9 in group A, 20 in group B) disclosed that belonging to group A constituted an independent factor for increased lymphocyte count (p < 0.001), body weight (p < 0.03), tricipital skinfold thickness (p < 0.01), middle-arm muscle circumference (p < 0.025), lean body mass (LBM) (p < 0.002), creatinine LBM related to body surface area (p < 0.001), and creatinine generation rate (p < 0.002). However, these data may have been biased by the high rate of noncompliance in group A. CONCLUSIONS: Protenplus proved to be unsuitable as a long term, oral protein-energy supplement in PD patients due to a high rate of noncompliance and intolerance, primarily among patients with lower residual renal function. The question of whether other products, better tolerated as nutritional supplements, could compensate for daily protein peritoneal losses in long-term PD remains open. PMID- 15796146 TI - Randomized, open label, controlled clinical trial of oral administration of an egg albumin-based protein supplement to patients on continuous ambulatory peritoneal dialysis. AB - BACKGROUND/AIM: Malnutrition is highly prevalent in patients on continuous ambulatory peritoneal dialysis (CAPD) and is a strong predictor of increased morbidity and mortality. Therefore, the aim of this study was to evaluate the effect of oral administration of an egg albumin-based protein supplement on the nutritional status of CAPD patients. METHODS: In this randomized, open label, controlled clinical trial, 28 CAPD patients were allocated to a study (n = 13) or a control (n = 15) group. Both groups received conventional nutritional counseling; the study group received, additionally, an oral egg albumin-based supplement. During a 6-month follow-up, all patients had monthly clinical and biochemical evaluations and quarterly assessments of adequacy of dialysis and nutrition. RESULTS: Serum albumin Levels were not different between groups; however, a significant increase (baseline vs final) was observed in the study group (2.64+/-0.35 vs 3.05+/-0.72 g/dL) but not in the control group (2.66+/-0.56 vs 2.80+/-0.54 mg/dL). Calorie and protein intake increased more in the study group (calories 1331+/-432 vs 1872+/-698 kcal; proteins 1.0+/-0.3 vs 1.7+/-0.7 g/kg) than in the control group (calories 1423+/-410 vs 1567+/-381 kcal; proteins 1.0+/-0.4 vs 1.0+/-0.3 g/kg). Similarly, non-protein nitrogen appearance rate (nPNA) increased significantly more in the study (1.00+/-0.23 vs 1.18+/-0.35 g/kg/day) than in the control group (0.91+/-0.11 vs 0.97+/-0.14 g/kg/ day). Triceps skinfold thickness (TSF) and midarm muscle area (MAMA) displayed a nonsignificant trend to a greater increase in the study group (TSF 16.7+/-8.7 vs 18.3+/-10.7 mm; MAMA 23.8+/-6.2 vs 25.8+/-5.9 cm2) than in controls (TSF 16.4+/ 5.7 vs 16.9+/-7.0 mm; MAMA 28.7+/-7.8 vs 30.0+/-7.9 cm2). At the end of follow up, the frequency of patients with moderate or severe malnutrition decreased 6% in the control group and decreased 28% in the study group. At the final evaluation, the most important predictors of serum albumin were the oral egg albumin-based supplement administration and protein intake (p < 0.05); secondary predictors (p = 0.06) were peritoneal transport rate and MAMA. CONCLUSIONS: In the study group, oral administration of the egg albumin-based supplement significantly improved serum albumin, calorie and protein intake, and nPNA, and, compared to controls, this maneuver was associated with a trend to increased anthropometric parameters and improved Subjective Global Assessment evaluation. Oral administration of the albumin supplement and protein intake were the most significant predictors of serum albumin at the end of follow-up. This oral supplement may be a safe, effective, and cheap method to improve nutritional status in peritoneal dialysis patients. PMID- 15796148 TI - Peritoneal catheter splicing for distant relocation of poorly selected exit sites. PMID- 15796147 TI - Determination of high and low molecular weight molecules of icodextrin in plasma and dialysate, using gel filtration chromatography, in peritoneal dialysis patients. AB - OBJECTIVE: The aim of this study was to apply high performance liquid chromatography (HPLC) with modern gel filtration media to determine high molecular weight (HMW) icodextrin fractions and low molecular weight (LMW) icodextrin metabolites in dialysate and plasma in peritoneal dialysis (PD) patients on treatment with icodextrin, and to explore the potential relationships between these compounds, alpha-amylase activity, and glomerular filtration rate. DESIGN: Retrospective study of dialysate and plasma samples from PD patients. SETTING: Samples were collected at one PD center. PATIENTS: Blood and dialysate samples were obtained from PD patients who were subdivided into three groups: patients using only glucose-based peritoneal dialysis fluid (GPDF; GLU group, n = 23), patients studied after the first long dwell with icodextrin-based peritoneal dialysis fluid (IPDF; 1st ICO group, n = 24), and patients who were regular users of IPDF for the long dwells (ICO group, n = 9). METHODS: LMW icodextrin metabolites [i.e., maltose (G2), maltotriose (G3), maltotetraose (G4), maltopentaose (G5), maltohexaose (G6), and maltoheptaose (G7)] and HMW fractions were determined in plasma and dialysate using two different gel filtration HPLC methods. Enzymatic hydrolysis with amyloglucosidase to glucose yielded the total carbohydrate content and this was used to validate the HPLC results. alpha Amylase activity was determined using a routine method. RESULTS: The results obtained by gel filtration HPLC yielded values of LMW metabolites and HMW fractions in plasma and dialysate in agreement with results obtained with enzymatic hydrolysis. HMW fractions were not detectable in plasma. Absorption of icodextrin from the peritoneal cavity during the long dwell (10 - 16 hours) was, on average, 39% of the amount instilled. During the long dwell, there was a relative decrease in the dialysate concentration of the largest HMW fractions (>21.4 kDa). Plasma concentration of the LMW icodextrin metabolites G2-G7 was highest in the ICO group (2.65+/-0.54 mg/mL) but also higher in the 1st ICO group (1.97+/-0.57 mg/mL) compared with the GLU group (0.52+/-0.23 mg/mL). Plasma alpha amylase activity was significantly lower in the 1st ICO group and in the ICO group compared with the GLU group. CONCLUSIONS: Accurate analysis of HMW icodextrin fractions in dialysate and LMW icodextrin metabolites in plasma and dialysate in PD patients can be achieved by gel filtration HPLC with two different columns. This method can be used to study the complex pattern of changes in icodextrin and its metabolites in plasma and dialysate. The finding that HMW icodextrin fractions were not detected in plasma was unexpected, and differs from results of previous studies by other researchers. PMID- 15796149 TI - Asymptomatic bowel perforation by a Tenckhoff catheter. PMID- 15796150 TI - Treatment of nodular parathyroid hyperplasia with percutaneous ethanol injection in a CAPD patient. PMID- 15796151 TI - Unusual presentation of peritonitis in a CAPD patient with lupus vasculitis. PMID- 15796152 TI - New anticancer agents: in vitro and in vivo evaluation. AB - The rapid emergence of new anticancer agents is a tremendous challenge for basic, pre-clinical and clinical research to evaluate and eventually integrate these new agents into clinical routine. Standardized, well-established in vitro and in vivo methods are available for the experimental evaluation of new anticancer agents. A step-wise procedure from in vitro to in vivo experiments using non-functional, functional non-clonogenic and, if applicable, clonogenic assays allows reduction of the number of promising agents for further clinical testing. PMID- 15796153 TI - Radiometallation of receptor-specific peptides for diagnosis and treatment of human cancer. AB - Radiolabeled, receptor-specific peptides are becoming increasingly popular as targeting vectors for the design and development of new diagnostic and therapeutic radiopharmaceuticals. The over-expression of functioning receptors on a variety of human cancers makes this method of drug development a viable tool for tumor targeting in vivo. This review describes some of the more recent efforts that are currently underway towards development of new receptor-specific radiopharmaceuticals. Diagnostic/therapeutic radionuclides, specific metal co ordinating ligands/chelating systems, spacer technology, radiolabeling protocols, and specific peptides/peptide conjugates will be discussed in detail. PMID- 15796154 TI - Phenolic compounds, sodium salicylate and related compounds, as inhibitors of tumor cell growth and inducers of apoptosis in mouse leukemia L1210 cells. AB - The effects of a series of phenolic compounds were compared to the effects of sodium salicylate (2-hydroxybenzoate) on the growth, cell cycle and apoptotic effects in wild-type (WT) and deoxyadenosine-resistant (Y8) L1210 leukemia cells. These compounds included: salicylaldehyde, salicylaldoxime, salicylhydroxamic acid, salicylamide, 5-aminosalicylate and 5-sulfosalicylate. The IC50 values for inhibition of tumor cell growth ranged from 40 microM for salicylaldhyde to greater than 4 mM for 5-sulfosalicylate. There appeared to be an excellent correlation between the IC50 value for a compound and the ratio of octanol/aqueous distribution. Salicylamide caused a G2/M block in both the WT and Y8 L1210 cells, while salicylalehyde caused a G0/G1 block in both the WT and Y8 cells. Salicylamide and salicylaldoxime caused a much greater apoptotic effect in the Y8 cells than in the parental WT L210 cells. These data suggest that salicylaldehyde and salicylaldoxime, the most active compounds in this series, may provide the lead chemicals from which other more active drugs can be synthesized. PMID- 15796155 TI - New anticancer agents: in vitro and in vivo evaluation of the antitumor and antimetastatic actions of various compounds isolated from medicinal plants. AB - In this review, in the search for the development of new anticancer drugs, the effects of compounds isolated from various medicinal plants on tumor growth and metastasis, using mice bearing a highly metastatic drug-resistant mouse tumor, were studied. The antitumor and antimetastatic actions of stilbene derivatives isolated from Polygonum and Cassia species were examined. Among the stilbene derivatives, resveratrol and cassiagrol A (stilbene dimer) displayed antitumor and antimetastatic actions through the inhibition of tumor-induced neovascularization in in vitro and in vivo models. It was found that two chalcone derivatives from Angelica keiskei roots also inhibited tumor growth and metastasis in tumor-bearing mice through the inhibition of tumor-induced neovascularization and/or the inhibition of immune suppression caused by tumors. Recently, basidiomycete fungi have been used for the treatment of cancer. Then, the low molecular weight substances were isolated from Agaricus blazei and Ganoderma lucidum as antitumor and antimetastatic substances. It is suggested that these substances of basidiomycete also inhibited tumor growth and metastasis to the lung through the inhibition of tumor-induced neovascularization and/or the inhibition of immune suppression caused by tumors. PMID- 15796156 TI - On the potential use of flavonoids in the treatment and prevention of pancreatic cancer. AB - Pancreatic cancer is a disease carrying a dismal prognosis, with overall 5-year survival at around 4%. Recent clinical trials of adjuvant therapies have not found a dramatic increase in median survival. In the current review, we examine the available literature on flavonoids, a group of naturally occurring substances, for their effects on cancer cells and potential for therapy of pancreatic cancer in the future. With the available in vitro and in vivo data, it is likely that flavonoids will move into the clinical arena as therapeutic or preventive tools for cancer. PMID- 15796157 TI - Synergistic antiproliferative action of the flavonols quercetin and kaempferol in cultured human cancer cell lines. AB - The consumption of vegetables containing the flavonols quercetin and kaempferol reduces the risk of cancer. We utilized human gut (HuTu-80 and Caco-2) and breast cancer cells (PMC42) to show the synergistic effect of quercetin and kaempferol in reducing cell proliferation. A trend in reduction of total cell counts was seen following a single exposure, a 4-day exposure or a 14-day exposure to quercetin and kaempferol. Combined treatments with quercetin and kaempferol were more effective than the additive effects of each flavonol. The reduction in cell proliferation was associated with decreased expression of nuclear proliferation antigen Ki67 and decreased total protein levels in treated cells relative to controls. In conclusion, the synergistic antiproliferative effect of quercetin and kaempferol demonstrated in cultured human cells has broad implications for understanding the influence of dietary nutrients in vivo, where anticancer effects may be a result of nutrients which act in concert. PMID- 15796158 TI - Clinical pharmacokinetics of imatinib mesylate. AB - This review presents the clinical pharmacokinetics of imatinib mesylate. Aspects regarding absorption, tissue distribution, elimination and kinetic interactions are also discussed. PMID- 15796159 TI - Review: potential role of cell cycle synchronizing agents in combination treatment modalities of malignant tumors. AB - Malignant neoplasms consist of heterogeneous cell populations and their cellular elements proliferate asynchronously. Since the tumor cells of various cell cycle phases respond differently to many chemotherapeutic drugs, attempts at synchronization seemed to be a promising way to achieve a more powerful antineoplastic effect. Mainly based on in vitro data, it was shown that numerous compounds, including hormones, were able to arrest the cell cycle in different phases, and some of them also induced apoptotic cell death. The better understanding of the molecular mechanisms of cell cycle control has brought the cyclin-dependent kinases into focus and hundreds of compounds have been synthesized in order to regulate malignant cells at their checkpoints, especially at G1 progression. Some of these compounds have been found to be effective not only in vitro, but also in in vivo experiments, and they were further evaluated in Phase I - II clinical trials. Generally speaking, these studies have yielded modest, although potentially promising, results, but the adverse effects sometimes restricted the applicability of the products. Nevertheless, extended studies in cancer patients are under way. Moreover, after encouraging preclinical investigations, the combination of cell cycle regulators with different cytostatic drugs may offer a novel therapeutic alternative in the field of oncology. PMID- 15796160 TI - Induction of apoptosis in human colon cancer HCT116 cells treated with an extract of the plant product, Chios mastic gum. AB - A hexane extract of the plant product Chios mastic gum (He-CMG) is demonstrated to kill human colon cancer cells in vitro via the process of anoikis. Specifically, the sequence of events includes He-CMG-induced GI-arrest of the cells, detachment of the cells from the substrate and subsequent apoptosis. Anoikis is dependent on the concentration and duration of treatment with He-CMG. Presence of the pan-caspase inhibitor, Z-VAD-fmk, did not prevent cell detachment, but it did prevent apoptosis of the detached cells indicating that the process of cell detachment, but not apoptosis, is independent of caspase activation. He-CMG-induced apoptosis is associated with activation of the initiator caspases-8, and -9 and the effector caspase-3. Caspases are activated in cells at a relatively long time after detachment, and caspase-3 activation may require caspase-8 or caspase-9 activation, as determined by using HCT116 isogenic clones impaired in apoptosis mechanisms that involve these two caspases. Finally, electron microscopy observations indicated a time-dependent appearance of morphological features both typical and non-typical of apoptosis in cells treated with He-CMG for various periods of time. Taken together, the results demonstrated that He-CMG induces apoptosis in HCT116 cells and, therefore, further in vivo and in vitro studies of the anticancer activities of this plant product are warranted. PMID- 15796161 TI - A safety study of oral tangeretin and xanthohumol administration to laboratory mice. AB - BACKGROUND: The detection of molecular targets for flavonoids in cell signalling has opened new perspectives for their application in medicine. Both tangeretin, a citrus methoxyflavone, and xanthohumol, the main prenylated chalcone present in hops (Humulus lupulus L.), act on the mitogen-activated protein kinase pathway and await further investigation for administration in vivo. MATERIALS AND METHODS: A safety study was designed in laboratory mice orally administered concentrates of purified tangeretin (1 x 10(-4) M) or xanthohumol (5 x 10(-4) M) at libitum for 4 weeks. Blood samples were collected for the analysis of a variety of haematological and biochemical parameters. RESULTS: A reduction of the circulating lymphocyte number was noticed for tangeretin, while all other parameters were unaffected by treatment with either tangeretin or xanthohumol. The parameters encompassed an integrity check of the following tissues and organs: bone marrow, liver, exocrine pancreas, kidneys, muscles, thyroid, ovaries and surrenal cortex. Furthermore, no differences were noted in the metabolism of proteins, lipids, carbohydrates and uric acid, as well as in ion concentrations. CONCLUSION: All data indicate that oral administration of tangeretin or xanthohumol to laboratory mice does not affect major organ functions and opens the gate for further safety studies in humans. PMID- 15796162 TI - Reactive oxygen species and redox-induced programmed cell death due to MK 886: cells ("soil") "trump" agent ("seed"). AB - Micromolar concentrations of the five-lipoxygenase inhibitor, MK 886 induce a "type 1" (apoptotic, extrinsic, death domain, receptor-dependent, caspase positive) form of programmed cell death in Bcl-2-positive U937 human monoblastoid and HL-60 myeloid leukemia cells. A "type 2" (intrinsic, mitochondria-dependent, autophagic, in some examples caspase-negative (Panc-1)) form is induced in Panc-1 pancreatic and PC3 prostate cell lines. The latter two lines from epithelial derived solid human cancers are Bcl-2-negative. Micromolar MK 886 induces an acute rise in Ca2+ in washed, Ca2+-poor U937 and HL60 cells in Ca2+ and Mg2+-free Hank's buffer. In U937 cells, much of the increase, or more properly redistribution, is nuclear in location (HL-60 not tested). No MK-886-induced acute Ca2+ increase developed in Panc-1 or PC3 cells. Bcl-2-positive HeLa cervical cancer cells exhibited an acute MK 886-induced increase in Ca2+. In the U937, PC3 and Panc-1 cells examined, MK-886 rapidly increased oxidative stress and decreased mitochondrial membrane potential, indicating that neither event is directly determinative for the altered distribution of Ca2+ or the form of PCD observed. Inhibition of increased U937 Ca2+ by the anti-oxidant, N-acetyl-L cysteine, the effects of inhibitors of mitochondrial function including antimycin A, atractyloside, cyclosporin A, the L/N channel blocker loperamide, the intracellular chelator BAPTA and 2 agents, HA-14 and 3-methyl-antimycin A3 that impair Bcl-2 function further define these events. These differences in the Ca2+ response and possibly also the form of PCD that results may depend upon the presence of Bcl-2 or a related protein participating in a juxta-nuclear / nuclear Ca2+ ion channel. The role of mitochondria, the mechanism by which increased oxidative stress initiates the rapid release of Ca2+ from intracellular, possibly juxta-nuclear / nuclear sites or its redistribution to U937 Ca2+ nuclei, and whether this "signal" or possibly even ROS themselves mandate the type of PCD observed, presumably by differential modulation of transcription, remain to be determined. Lastly, these results demonstrate that, as might be expected, "soil" (cell type) trumps "seed" (inciting agent)". PMID- 15796163 TI - Re-evaluation of cytotoxicity and iron chelation activity of three beta-diketones by semiempirical molecular orbital method. AB - We investigated the chelating effect of FeCl3 on three beta-diketones, curcumin [1], (-)-3-(trifluoroacetyl)camphor [2] and 3-formylchromone [3], as judged by changes in their cytotoxicity and absorption spectra. Addition of an equimolar concentration of FeCl3 almost completely abrogated the cytotoxicity and changed the pattern of absorption spectra (decrease in the peak height at 430 nm) of [1], without affecting those of [2] and [3]. A semiempirical molecular orbital method (CAChe), based on these experimental data, proposed the estimated structure of [1] chelated with FeCl3 at 1:1 molar ratio. The present study suggests that [1], but not [2] and [3], is an iron chelator. PMID- 15796164 TI - 4-[3,5-Bis(trimethylsilyl)benzamido] benzoic acid (TAC-101) induces apoptosis in colon cancer partially through the induction of Fas expression. AB - BACKGROUND: 4-[3,5-Bis (trimethylsilyl) benzamido] benzoic acid (TAC-101) is a novel retinobenzoic acid derivative, which has a specific binding affinity to the retinoic acid receptors (RAR)-alpha and -beta. Apoptotic induction by TAC-101 was investigated using a rat hepatic metastatic model of rat RCN-9 colon cancer cells in vivo and FACScan analysis with the DLD-1 human colon cancer cell line in vitro. MATERIALS AND METHODS: Hepatic metastatic tumors were induced using intra portal injection of RCN-9 cells into F344 rats in vivo. TAC-101 (8 mg/kg) was orally administered for 5 consecutive days a week for 4 weeks. Subsequently, hepatic tumors were counted after laparotomy. Apoptotic index (A.I.) in the hepatic tumors was evaluated using immunohistochemistry for single-stranded DNA. The proliferative index (P.I.), Fas and Fas ligand were also immunohistochemically evaluated. Moreover, evaluation of apoptosis by TAC-101 in vitro using FACScan analysis was performed in the DLD-1 human colon cancer cell line. RESULTS: Oral administration of TAC-101 resulted in a significant inhibition of hepatic metastasis without weight loss of the rats. TAC-101 significantly decreased P. I. but increased A. I. in the hepatic metastatic tumors. TAC-101 did not affect the expression of Fas ligand, but obviously increased the expression of Fas in the metastatic tumors. Moreover, TAC-101 induced early apoptosis in DLD-1 cells in a time-dependent manner in vitro. CONCLUSION: These findings suggest that TAC-101 inhibits hepatic metastasis of colon cancer and induces apoptosis partially through enhanced Fas expression. PMID- 15796165 TI - Antimetastatic activity of a synthetic serine protease inhibitor, FOY-305 (Foypan). AB - Metastasis is one of the major causes of mortality in cancer. It is well known that the activities of cell surface serine proteases are especially enhanced in malignant tumors. Proteolytic degradation of the extracellular matrix and basal membrane is a crucial event for tumor cell invasion and metastasis formation. FOY 305 (Foypan), a remedy for tumor pancreatitis, is a broad spectrum synthetic serine protease inhibitor which inhibits enzymatic activities including trypsin, thrombin, kallikrein and plasmin. Using Lewis lung carcinoma cell, we found that FOY-305 inhibited both spontaneous and experimental pulmonary metastasis. Furthermore, the combined treatment of FOY-305 and a traditional anti-cancer drug, 5-FU or bleomycin, resulted in marked enhancement of anti-pulmonary metastatic activity. PMID- 15796166 TI - Effect of cordycepin (3'-deoxyadenosine) on hematogenic lung metastatic model mice. AB - We investigated the anti-metastatic effect of cordycepin (3'-deoxyadenosine) on a hematogenic metastatic mouse model which was intravenously injected with B16-BL6 melanoma cells. A 3-hour exposure to various concentrations of cordycepin (0.3, 1 and 3 microg/ml) dose-dependently reduced the number of nodules formed in lung at 15 days after the tumor injection. To elucidate the mechanism of this anti metastatic effect, we examined the effect of cordycepin on the invasiveness of B16-BL6 cells using a chemo-invasion chamber in vitro. The B16-BL6 cells pretreated with cordycepin (3 microg/ml) for 3 hours showed a significant decrease in invasiveness. Under the same conditions, however, cordycepin did not influence the growth curve of B16-BL6 cells at concentrations up to 3 microg/ml. These results suggest that cordycepin exerts an anti-metastatic action, in part, by inhibiting the invasiveness of mouse melanoma cells. PMID- 15796167 TI - Effects of ellagic acid by oral administration on distribution and metabolism of 2-aminofluorene in Sprague-Dawley rats. AB - The effects of ellagic acid on the in vivo N-acetylation and metabolism of 2 aminofluorene (2-AF) were investigated in bladder, blood, colon, kidney, liver, feces and urine samples from male Sprague-Dawley rats. Major metabolites such as 1-OH-2-AAF, 8-OH-2-AAF and 9-OH-2-AAF were found in bladder tissues, 1-OH-2-AAF, 5-OH-2-AAF and 8-OH-2-AAF were found in blood samples, 1-OH-2-AAF, 3-OH-2-AAF, 5 OH-2-AAF, 8-OH-2-AAF and 9-OH-2-AAF were found in colon tissues, 1-OH-2-AAF, 3-OH 2-AAF and 9-OH-2-AAF were found in kidney tissues, 1-OH-2-AAF, 3-OH-2-AAF and 8 OH-2-AAF were found in liver tissues, 1-OH-2-AAF, 3-OH-2-AAF, 5-OH-2-AAF and 8-OH 2-AAF were found in feces samples and 1-OH-2-AAF, 3-OH-2-AAF, 5-OH-2-AAF and 8-OH 2-AAF were also found in urine samples after rats had been orally treated with 2 AF (50 mg/kg) for 24 h. Pretreatment of male rats with ellagic acid (10 mg/kg) 24 h prior to the administration of 2-AF (50 mg/kg) resulted in absence of 8-OH-2 AAF in bladder tissues, and there were significant decreases of 8-OH-2-AAF in blood and urine samples. In blood samples, amounts of 2-AAF and 8-OH-2-AAF were significantly decreased; in colon tissues, amounts of 2-AF, 1-OH-2-AAF and 3-OH-2 AAF, in liver tissues, amounts of 2-AAF, 1-OH-2-AAF and 3-OH-2-AAF, and in urine samples, amounts of 2-AF and 8-OH-2-AAF were significantly decreased in 24-h ellagic acid (EA)-treated rats before 2-AF was added to the diet. However, significantly increased 1-OH-2-AAF concentrations were found in urine samples in 24-h EA-treated rats before 2-AF was administered. In the EA and 2-AF rats, in the same time treated groups, bladder, colon and liver tissues, and feces and urine samples showed significant differences when compared to the ones without EA co-treatment. We saw significant decreases of the amounts of 2-AF and 1-OH-2-AAF in colon tissues. The feces samples showed increased amounts of 2-AAF in EA- and in 2-AF- treated rats in the same time groups, but urine samples showed a decreased amount of 8-OH-2-AAF in both EA-treated groups. The total amounts of 2 AF metabolites in bladder, blood, kidney and liver tissues showed significant difference between control and the group which was EA-treated 24 h before 2-AF was added. The total amounts of 2-AF metabolites in the liver, feces and urine showed significant decreases between control and EA-treated at the same time with 2-AF groups. This is the first report of EA affecting the N-acetylation and metabolism of 2-AF in rat tissues in vivo. PMID- 15796168 TI - Cytotoxic activity of styrylchromones against human tumor cell lines. AB - A total of 6 newly-synthesized styrylchromones (SC-1 approximately SC-6) were compared for their cytotoxic activity against three normal oral human cells (gingival fibroblast HGF, pulp cell HPC, periodontal ligament fibroblast HPLF) and four human tumor cell lines (squamous cell carcinoma HSC-2, HSC-3, submandibular gland carcinoma HSG, promyelocytic leukemia HL-60). All compounds showed higher cytotoxic activity against tumor cell lines than against normal cells. Among the 6 compounds, SC-3, SC-4 and SC-5, which have one to three methoxy groups, showed higher tumor specificity and water solubility. The cytotoxic activity of SC-3 and SC-5 was slightly reduced by a lower concentration of NADH, a quinone reductase, but that of SC-3 was enhanced by higher concentrations of NADH, possibly due to demethylation of the methoxy groups. Agarose gel electrophoresis demonstrated that SC-3 and SC-5 induced intemucleosomal DNA fragmentation in HL-60 cells and production of large DNA fragment in HSC-2 cells. Both SC-3 and SC-5 enhanced the enzymatic activity to cleave the substrates for caspases 3, 8 and 9, suggesting the activation of both extrinsic and intrinsic apoptosis pathways. ESR spectroscopy showed that these compounds produced no detectable amount of radical and did not scavenge superoxide anion generated by the hypoxanthine-xanthine oxidase reaction. The highly tumor-specific cytotoxic action and apoptosis-inducing capability of SC-3 and SC-5 suggest their applicability for cancer chemotherapy. PMID- 15796169 TI - Inhibition by Rikko-san and its major ingredients of LPS-stimulated nitric oxide production by mouse macrophage-like cells. AB - Rikko-san and its ingredients were investigated for their activity to modify nitric oxide (NO) production by unstimulated and lipopolysaccharide (LPS) stimulated mouse macrophage-like Raw 264.7 cells. LPS significantly stimulated the NO production by Raw 264.7 cells, and Rikko-san effectively inhibited the stimulation effect of LPS even at non-cytotoxic concentrations. Among 5 Rikko-san ingredients, Kanzo showed a similar magnitude of inhibition of NO production. Shoma was also slightly inhibitory. On the other hand, Ryutan, Saishin and Bofu did not show such a clear-cut stimulation effect, due to the co-existence of both inhibitory and stimulatory substance(s) for NO production. Thus NO stimulators were present in Rikko-san and its four ingredients except for Kanzo. Western blot analysis demonstrated that LPS induced the production of inducible NO synthase (iNOS), and that non-cytotoxic concentrations of Rikko-san and Kanzo significantly inhibited the LPS-stimulated iNOS expression. ESR spectroscopy showed that Rikko-san, Kanzo, Shoma and Saishin, but not Ryutan and Bofu, produced radical(s) under alkaline condition. All samples scavenged superoxide (produced by hypoxanthine-xanthine oxidase reaction) and NO (produced by 1 hydroxy-2-oxo-3-(N-3-methyl-3-aminopropyl)-3-methyl-1-triazene (NOC-7)), possibly by their general reducing activity. These data suggest that the inhibition of NO production by Chinese medicines investigated here may be the result of both the inhibition of iNOS expression and their radical scavenging activity. PMID- 15796170 TI - Antitumor effect of geranylamine derivatives on human hepatoma. AB - In this study, we examined the antitumor activities of isoprenoid derivatives conjugated with substrates of energy metabolism in human hepatoma-bearing athymic mice. Among these compounds, N-geranylpyruvic amide, N-geranyl-p pyruvaminobenzoic amide, N,N'-digeranylmalic diamide and N,N'-digeranyl-O acetylmalic diamide had strong antitumor effects. These geranylamine derivatives also inhibited in vitro cell growth. Sugar conjugates of geranylamine, geranic acid and mevalonic acid did not show any antitumor effect in vivo or in vitro. Although the geranylamine derivatives had no impact on the cell cycle distribution at 24 h, a sub-G1 (apoptotic) peak of varying magnitude was seen in DNA histograms of cells treated with the derivatives for 48 h. However, the geranylamine derivatives did not inhibit protein isoprenylation, which has been reported in cancer cells treated with several natural isoprenoids. These results suggest that the geranylamine derivatives conjugated with malic acid and pyruvic acid have a different mechanism of antitumor activity from that of natural isoprenoids. PMID- 15796171 TI - In vivo antitumor effect of ascorbic acid, lysine, proline and green tea extract on human prostate cancer PC-3 xenografts in nude mice: evaluation of tumor growth and immunohistochemistry. AB - BACKGROUND: Matrix metalloproteinases (MMPs), vascular endothelial growth factor (VEGF), Ki 67 (proliferative protein) and constituents of ECM play a critical role in angiogenesis, and are crucial in neoplastic invasion and metastasis. Based on the antitumor properties of certain nutrients, we investigated the effect of a diet containing lysine, proline, arginine, ascorbic acid and green tea extract on the growth of tumors induced by implanting human prostate cancer PC-3 cells in athymic nude mice and on the expression of MMPs, VEGF, Ki 67 and fibronectin in these tumors, as well as the production of mucin (by PAS staining). MATERIALS AND METHODS: Male nude mice (n =12) were inoculated with 3x10(6) prostate cancer PC-3 cells and randomly divided into two groups; Group A was fed a regular diet and Group B was fed a regular diet supplemented with 0.5% of the nutrient mixture (NM). Four weeks later, tumors were excised, weighed and processed for histology. RESULTS: The results showed inhibition of tumor growth in Group B. Histological studies revealed inhibition of MMP-9 and VEGF secretion and mitosis in Group B tissues. CONCLUSION: Nutrient supplementation strongly suppressed the growth of tumors without any adverse effects in nude mice, suggesting strong potential as an anticancer agent. PMID- 15796173 TI - Antitumour activity of Angelica archangelica leaf extract. AB - BACKGROUND: The purpose of this study was to examine the effect of a leaf extract from A. archangelica on the growth of Crl mouse breast cancer cells in vitro and in vivo. MATERIALS AND METHODS: The antiproliferative activity of the extract was measured by 3H-thymidine uptake in the Crl cells in vitro. Twenty mice were injected with the Crl cells, and 11 of them were fed A. archangelica leaf extract, and the progress of the tumours was followed. RESULTS: The leaf extract was mildly antiproliferative on the Crl cells with an EC50 of 87.6 microg/ml The antitumour activity of the extract was expressed in the mice by marked reduction in tumour growth. In the experimental animals, 9 out of 11 mice developed no or very small tumours, whereas control animals, not receiving the extract, developed significantly larger tumours (p<0.01), as estimated by Mann-Whitney U-test. The antitumour activity of the leaf extract could not be explained by the antiproliferative activity of furanocoumarins present in the extract. CONCLUSION: The results demonstrate the antiproliferative activity in vitro and antitumour activity in vivo of a leaf extract from A. archangelica PMID- 15796172 TI - The hepoxilin analog, PBT-3, inhibits growth of K-562 CML solid tumours in vivo in nude mice. AB - PBT-3 is one of a family of stable chemical analogs of the hepoxilins, products derived from arachidonic acid. We previously showed that PBT-3 caused apoptosis in the chronic myelogenous leukemia (CML) cell line K-562 in vitro (Anti-cancer Res 23: 3617-3622, 2003). It was as effective as Gleevec, a novel agent that blocks tyrosine kinase activity during treatment of CML. We describe, herein, the growth inhibiting effects of PBT-3 in nude mice into which K-562 cells were transplanted subcutaneously. Groups of mice were treated with vehicle as control, or PBT-3, or Gleevec. PBT-3 was effective during the 8-day treatment protocol in inhibiting the growth of the tumours in vivo as was Gleevec. Analysis of the tumours demonstrated the presence of apoptosis (DNA laddering and TUNEL assay) in both the PBT-3- and Gleevec-treated groups. These results demonstrate that PBT-3 is effective in vivo in controlling tumour growth and provides a novel platform for the therapeutic control of cancer. PMID- 15796174 TI - Ellagic acid inhibits growth and arylamine N-acetyltransferase activity and gene expression in Staphylococcus aureus. AB - It is well documented that N-acetyltransferase (NAT) plays a key role in the N acetylation of arylamine compounds. Ellagic acid was demonstrated to elicit dose dependent bacteriostatic activity and inhibition of N-acetylation of 2 aminofluorene (AF). N-acetylation of AF in S. aureus was determined by high preformance liquid chromatography. The apparent values of Km and Vmax of NAT were decreased after co-treatment with 0.5 mM ellagic acid in the cytosol of S. aureus. PCR also indicated that ellagic acid inhibited NAT gene expression (NAT mRNA) in S. aureus. PMID- 15796175 TI - Possible adjuvant cancer therapy by two prebiotics--inulin or oligofructose. AB - Dietary treatment with inulin or oligofructose incorporated in the basal diet for experimental animals: (I) reduced the incidence of mammary tumors induced in Sprague-Dawley rats by methylnitrosourea; (II) inhibited the growth of transplantable malignant tumors in mice; (III) decreased the incidence of lung metastases of a malignant tumor implanted intramuscularily in mice. (IV) Moroever, besides such cancer risk reduction effects, dietary treatment with inulin or oligofructose significantly potentiated the effects of subtherapeutic doses of six cytotoxic drugs commonly utilized in human cancer treatment. (V) The same prebiotics potentiated the effects of radiotherapy on solid form of TLT tumors to a statistically very high level. Such dietary treatment, with the inulin or oligofructose potentiating the effects of cancer therapy, might be introduced into classic protocols of human cancer treatment as a new, non-toxic and easily applicable adjuvant cancer therapy without any additional risk to patients. PMID- 15796176 TI - Heterodinucleoside phosphates of 5-fluorodeoxyuridine and arabinofuranosylcytosine--new drugs in cancer chemotherapy? AB - The incidence of cancer is rapidly increasing and malignancies have become the number two cause of deaths in the Western world after cardiovascular diseases. In particular, colon cancer represents one of the most frequent types of malignancy. Chemotherapy is, in addition to surgery and irradiation, still one of the main treatment options against this group of diseases. Here, several chemotherapeutic treatment modalities and anticancer compounds for the treatment of colon cancer are reviewed. In particular, a newer group of heterodinucleoside phosphates (dimers), consisting of two well known antimetabolites (5-FdUrd (5 Fluorodeoxyuridine) and Ara-C (Cytarabine)), are presented. These dimers were evaluated in several studies and might offer an additional option for the treatment of various malignancies, in particular colon carcinomas. The results are summarized in detail, as these dimers might have some significant advantages when compared with conventional regimens; they might be administered orally and might constitute an alternative treatment option for resistant tumors. PMID- 15796177 TI - 5-fluorouracil as a phosensitiser. AB - 5-FU exhibits a high fluorescence after irradiation with UV-vis light. An enhancement of the cytostatic activity of 5-FU under UV-vis irradiation was observed on an in vivo experimental model. PMID- 15796178 TI - Differential influence of pre- and post-delta-6-desaturation n6 polyunsaturated fatty acids on fibroblasts in culture, compared to n9 monounsaturated and n3 polyunsaturated fatty acids. AB - There is now considerable evidence that polyunsaturated fatty acids (PUFA) are effective in vitro at limiting the growth of cancer cells while not affecting normal cells to the same extent. Twenty carbon PUFA, especially of the n3 series, have been shown to be particularly potent; while the eighteen carbon n6 fatty acid, linoleic acid has been implicated in growth stimulation of breast cancer. We report here on the comparative effects of a range of eighteen and twenty carbon fatty acids of varying degrees of unsaturation on normal and transformed fibroblasts in culture. All moieties of the n3 series showed high potency in limiting transformed cell growth, while cis and trans monounsaturates and pre delta-6-desaturation n6 polyunsaturates induced a mixed response, even inducing comparative growth stimulation with some fatty acid concentrations. PMID- 15796180 TI - Anti-tumor activities of four chelating agents against human neuroblastoma cells. AB - BACKGROUND: Iron deprivation may be a therapeutic strategy for cancer. It can be achieved by using iron chelators. In this investigation, anti-neuroblastoma activities of a novel ferric chelator 2LL together with DFO, EDTA and DTPA were evaluated. MATERIALS AND METHODS: SH-Sy5y cells were cultured at 37 degrees C in 5% CO2/95% air in DMEM containing 10% fetal bovine serum. The cells were seeded in 96-well microtiter plates overnight. Then, chelating agents were added into the wells. After 48-hour incubation, viabilities were measured using the MTT method. RESULTS: DTPA had an IC50 value between 60-100 microM; DFO produced about 40% inhibiting effect at 150 microM; 2LL and EDTA displayed about 10% inhibiting effect at high concentrations. CONCLUSION: For SH-Sy5y cells, DTPA showed the strongest inhibiting effect, DFO displayed a moderate inhibiting effect, while 2LL and EDTA produced minor inhibition. To develop iron chelators as powerful anti-cancer agents is still a challenging task. PMID- 15796179 TI - Glutathione-related enzymes contribute to resistance of tumor cells and low toxicity in normal organs to artesunate. AB - The anti-malarial artesunate (ART) also inhibits the growth of cancer cells. The active moiety is an endoperoxide bridge whose cleavage generates reactive oxygen species and free radicals. We analyzed whether glutathione-related enzymes contribute to tumor resistance to ART and to the low toxicity of ART towards normal organs. The microarray-based mRNA expression of dihydrodiol dehydrogenase, gamma-glutamylcysteine synthase (gamma-GCS), glutathione S-transferases GSTM4, GSTT2, GSTZ1, and microsomal glutathione S-transferase MGST3 showed significant relationships (p <0.05) to cellular response to ART in 55 cell lines of the National Cancer Institute, USA. A tendency for correlation (0.05 or =20/40. The BSCVA remained the same or improved in 100% of eyes. After 4 months, 69.1% (47/68) of eyes in group 1 and 52% (13/25) of eyes in group 2 were within +/-1.00 D of the desired refraction; the mean refraction was stable between 4 and 24 months. Of the intraoperative complications, 69.2% were observed in the first 25 lenses implanted; postoperative complications included iris atrophy in 11.8% (11/93), lens decentration in 5.4% (5/93), and night glare in 6.4% (6/93) of eyes. No IOLs were removed. Mean endothelial cell loss was 2.8% at 4 months, 3.9% at 12 months, and 5.4% at 24 months. CONCLUSIONS: Our results regarding implantation of the Artisan phakic IOL confirm that these lenses are safe and effective for the correction of high myopia, with a stable refractive outcome but with a higher than normal rate of endothelial cell loss during 2-year follow-up. PMID- 15796216 TI - A test eye for wavefront eye refractors. AB - PURPOSE: A multi-site study was conducted to test feasibility of a modified automatic refractor style test eye as a test device for wavefront refractors of various types and to determine whether a) they could be measured and b) when measurements could be made, to see if they were similar. This study did not attempt to assess which instrument most accurately measures the aberrations of the test eye or human eye. METHODS: Three automatic refractor style test eyes were modified for use as test devices for wavefront refractors. One had a simple spherical front surface, and two had additional aberrations added. The test eyes and holder were circulated to 11 test sites where attempts were made to measure them with eight different wave-front refractor systems. RESULTS: Eight (100%) of the eight wavefront refractor systems tested successfully measured the test eyes. The systems did not give similar results for the same test eye. In some cases, coma was reported where none was present. Differences in reported defocus values reflect different approaches for compensating for the dispersion of the eye. A corneal topography system could measure and recognize the aberrations of the test eyes as well as the wavefront refractor systems tested. Interferometry, on the other hand, did not prove to be a successful method to assess the surface of the test eyes. CONCLUSIONS: The test eye design may be used as a test device for wavefront refractor systems. This type of test eye can detect systematic differences between various wavefront refractors and can serve as a useful calibration and comparison tool. PMID- 15796217 TI - Effect of optical zone ablation diameter on LASIK-induced higher order optical aberrations. AB - PURPOSE: To compare the postoperative higher order aberrations among eyes that had received conventional laser in situ keratomileusis (LASIK) treatment at different optical zone diameters using the Alcon LADARVision 4000 platform (Alcon, Ft Worth, Tex). METHODS: Monochromatic wavefront aberrations at 6.5-mm pupil size of 96 eyes of 96 Hong Kong Chinese patients with normal myopic astigmatism were objectively measured using the Alcon LADARWave aberrometer before and after (at least 6 months) conventional LASIK with the Autonomous LADARVision 4000 excimer laser. Patients were divided into three groups according to optical zone diameter--group 1 (n=32), optical zone diameter <6.5 mm; group 2 (n=32), optical zone diameter 6.5 mm; and group 3 (n=32), optical zone diameter >6.5 mm). RESULTS: Mean optical zone diameters of groups 1, 2, and 3 were 6.2+/ 0.1 mm, 6.5+/-0.0 mm, and 6.9+/-0.2 mm, respectively. Total higher order, coma, and spherical aberrations before LASIK were 0.67+/-0.27 microm, 0.27+/-0.17 microm, and 0.41+/-0.14 microm in group 1; 0.56+/-0.26 microm, 0.28+/-0.15 microm, and 0.32+/-0.11 microm in group 2; and 0.62+/-0.25 microm, 0.34+/-0.15 microm, and 0.37+/-0.09 microm in group 3. After surgery, the aberrations increased to 1.20+/-0.21 microm, 0.71+/-0.25 microm, and 0.70+/-0.19 microm in group 1; 0.97+/-0.18 microm, 0.47+/-0.22 microm, and 0.69+/-0.13 microm in group 2; and 0.91+/-0.20 microm, 0.41+/-0.22 microm, and 0.56+/-0.12 microm in group 3. Compared to group 1, groups 2 and 3 had significantly less coma aberrations after LASIK (P=.03); the total high order aberration in group 3 was also significantly lower than in group 1 (P=.04) after treatment. CONCLUSIONS: In conventional LASIK treatment using the Alcon LADARVision 4000 platform, a larger surgical optical zone diameter significantly decreases higher order aberrations after LASIK. PMID- 15796218 TI - Involvement of neural adaptation in the recovery of vision after laser refractive surgery. AB - PURPOSE: To investigate whether the gradual improvement in unaided visual acuity commonly seen after laser refractive surgery is attributable, in part, to neural adaptation to blur. METHODS: Unaided logMAR visual acuity was measured at presentation (blur adapted) and immediately after refraction and removal of refractive correction (correction adapted) in 26 patients with low myopic residual refractive error after laser in situ keratomileusis (LASIK). The difference in unaided visual acuity (correction adapted--blur adapted) indicates a dominance of practice effect (if negative) or blur adaptation (if positive). A combination of blur adaptation and practice effect is possible. RESULTS: Overall, the mean unaided visual acuity at presentation (blur adapted) was 0.16+/-0.16 (mean+/-standard deviation) logMAR, and the mean unaided visual acuity immediately after refraction and removal of refractive correction (correction adapted) was 0.14+/-0.14 logMAR, giving a difference (correction adapted--blur adapted) of -0.02+/-0.06 logMAR. This difference was not significant (analysis of variance [ANOVA] F1,25=0.204, P>.05), suggesting neither blur adaptation nor practice effect. However, during the first 10 weeks after surgery, the difference in unaided visual acuity was -0.07+/-0.05 logMAR, suggesting a practice effect. After 10 weeks, the mean difference was +0.02+/-0.05 logMAR, suggesting any practice effect is offset by blur adaptation. These values were significantly different (ANOVA F1,25= 13.53, P<.01). CONCLUSIONS: These data suggest that patients do not adapt to surgically induced blur, on average, until 10 weeks after LASIK. The reason for this delay is uncertain; perhaps instability of blur hinders adaptation during the early postoperative period. Part of the gradual visual improvement after LASIK appears to be due to neural adaptation to blur. PMID- 15796219 TI - Alcohol and mechanical scraping for epithelial ingrowth following laser in situ keratomileusis. AB - PURPOSE: To determine whether the use of 70% isopropyl alcohol is useful as an adjunct to simple mechanical removal of epithelial ingrowth following laser in situ keratomileusis (LASIK). METHODS: A retrospective comparison of two techniques of epithelial ingrowth removal following LASIK was performed. A total of 38 procedures on 31 patients were included. In one group, the epithelium was removed using a simple mechanical scraping technique and in the other, a similar technique combined with the application of 70% isopropyl alcohol was used. The rates of recurrence in the two groups were compared using a chi-square analysis. RESULTS: The total incidence of epithelial ingrowth recurrence in both groups was 57.9% (22/38). The incidence of clinically significant ingrowth recurrence in both groups was 34.2% (13/38). In the scrape group, the recurrence of any ingrowth was 43.8% (7/16) and the recurrence of clinically significant ingrowth was 31.3% (5/16). In the alcohol group, these rates were 68.2% (15/22) and 36.4% (8/22), respectively. The difference between the two groups did not show statistical significance in either comparison (P=.14 and P=.75, respectively). CONCLUSIONS: The use of 70% isopropyl alcohol showed no additional benefit over simple mechanical scraping in the removal and prevention of epithelial ingrowth recurrence following LASIK. PMID- 15796220 TI - Use of polymyxin as an endotoxin blocker in the prevention of diffuse lamellar keratitis in an animal model. AB - PURPOSE: To determine whether bacterial endotoxin, lipopolysaccharide (LPS), could induce diffuse lamellar keratitis (DLK) in an animal model and whether DLK could be prevented by endotoxin blockers such as polymyxin. METHODS: Laser in situ keratomileusis (LASIK) flaps were created in rabbit eyes. The stromal bed was treated with 20 microg of Burkholderia cepacia LPS or balanced salt solution (BSS). Development of DLK, histological degree of inflammation, and presence of LPS detected by anti-LPS antibody were evaluated after 48 hours. In a second experiment, all eyes received LPS and were randomly assigned to receive either polymyxin in the form of two drops of Polytrim (Allergan, Irvine, Calif) on the stromal bed or two drops of BSS. RESULTS: In the animal model study, LPS was significantly associated with the development of DLK (P<.05, n=30). Infiltration with polymorphonuclear cells and presence of DLK were found in LPS treated eyes but not in controls. In the second experiment, 4 (27%) of 15 eyes that received polymyxin in addition to LPS developed DLK compared to 18 (95%) of 19 eyes that received only LPS (P<.05, n=34). There was a trend towards higher flap displacement in polymyxin treated eyes but this was not significant (P=.07). CONCLUSIONS: Diffuse lamellar keratitis in a rabbit model can be caused by bacterial endotoxin (LPS). Endotoxin blockers, such as polymyxin, are effective in decreasing the incidence of endotoxin-induced DLK in a rabbit model. PMID- 15796221 TI - Effect of a yellow filter on contrast sensitivity and disability glare after laser in situ keratomileusis under mesopic and photopic conditions. AB - PURPOSE: To analyze the effect of a yellow filter on contrast sensitivity and disability glare under mesopic and photopic luminance conditions in laser in situ keratomileusis (LASIK) patients and control subjects. METHODS: Contrast sensitivity with and without glare was determined in 27 patients who had undergone LASIK at least 1 year previously and in 30 control subjects. Tests were performed with and without a coated yellow filter (X-482 nm cut-off) using the Mesotest II or Mesoptometer II (Oculus, Wetzlar, Germany) under mesopic conditions, and the Contrast Glaretester 1000 (Takagi, Seiko Co Ltd, Nagano, Japan) under photopic conditions. RESULTS: Under mesopic conditions, log contrast sensitivity without glare decreased by 0.14 log units in the LASIK group. When the yellow filter was used, this variable showed a significant increase of 0.04 log units and the proportion of patients able to discriminate the mesopic contrast limit of 1:5 rose from 70% to 78%. With glare, the yellow filter also improved contrast sensitivity in LASIK patients, but not significantly. Under photopic conditions, no statistical differences were observed between results obtained with and without the yellow filter in the LASIK group or between the LASIK and control group without glare. CONCLUSIONS: Mesopic contrast sensitivity without glare was worse in LASIK patients and increased significantly with the yellow filter. The filter had no effect under photopic conditions. No disability glare differences were noted between the LASIK and control groups or between the LASIK without and with filter groups under mesopic and photopic conditions. PMID- 15796223 TI - Intraocular pressure measurements after conductive keratoplasty. AB - PURPOSE: To determine the possible impact of conductive keratoplasty (CK) on intraocular pressure (IOP) measurements. METHODS: A prospective, single-center, noncomparative interventional case series was performed. Baseline and postoperative IOPs were measured by Goldmann applanation tonometry in 32 eyes of 18 patients who underwent CK for hyperopia correction. Mean follow-up was 11.9 months (range: 8 to 18 months). RESULTS: After CK, a statistically significant decrease in the measured IOP was observed (before CK: 14.22+/-1.64 vs after CK: 12.66+/-2.21, P<.001). The change in IOP readings postoperatively was not correlated with age, sex, keratometric readings, or attempted correction. CONCLUSIONS: Despite the limitations due to the small number of patients enrolled in this study, the applanation tonometer appears to underestimate the true IOP after CK. PMID- 15796222 TI - Visual acuity and corneal topographic changes related with pterygium surgery. AB - PURPOSE: Applied pre- and postoperative corneal topography outcomes with visual acuities were evaluated to determine the surgical results in pterygium therapy. METHODS: The study group consisted of 30 eyes of 26 patients with primary and recurrent pterygium. Mean patient age was 52.26+/-11.50 years (range: 27 to 68 years). Pre- and postoperative visual acuity and comeal topography were evaluated for each case. Statistical analysis was performed using the repeated-measure test. RESULTS: The mean uncorrected visual acuity was 0.41+/-0.30 preoperatively and 0.63+/-0.26 postoperatively (P<.001). The mean best spectacle-corrected visual acuity was 0.59+/-0.28 preoperatively and 0.84+/-0.22 postoperatively (P<.001). Surface regularity index ranged from 1.96+/-1.08 preoperatively to 1.09+/-0.76 postoperatively (P<.001). The mean surface asymmetry index was 3.05+/ 2.85 preoperatively and 1.39+/-1.70 postoperatively (P=.003). The mean topographic astigmatism was 4.65+/-3.02 preoperatively and 2.33+/-2.26 postoperatively (P=.003). CONCLUSIONS: Examination of the topographic records reveals pterygium-associated corneal flattening in the horizontal meridian along the line of the pterygium. The improvement in topographic pattern and best spectacle-corrected visual acuity can be used as one indicator of the success of pterygium surgery. PMID- 15796224 TI - Theoretical elastic response of the cornea to refractive surgery: risk factors for keratectasia. AB - PURPOSE: To explore the role that mechanical elastic factors may have in post refractive surgery corneal phenomena, from mild curvature changes to keratectasia. METHODS: The central cornea near the apex was modeled as an elastic spherical thin shell loaded by the intraocular pressure (IOP). Equations for myopic laser in situ keratomileusis (LASIK) were obtained to estimate shifts and curvature changes of the posterior corneal surface at the apex. The effect of every parameter was studied, identifying potential risk factors for ectasia. RESULTS: Theoretically, corneal thinning by ablation will produce an elastic deformation of the posterior surface that depends on the corneal parameters (curvature, Young's modulus, Poisson ratio, and thickness), the IOP, and the ablation profile. In particular, a forward shift and an increase in power of the posterior surface was predicted for myopic LASIK, in agreement with previous experimental findings. These changes rise non-linearly with the attempted correction, and are greater for thinner preoperative corneas, higher IOP smaller Young's modulus, and thicker flaps. Corneas with Young's modulus half the average or less, or thickness <500 microm, may present high risk of ectasia, especially for high IOP and thick flaps. CONCLUSIONS: Some postoperative effects may be explained in part by elasticity. Research efforts are needed to explain other biomechanical behaviors. The accepted criterion of 250 microm residual bed is insufficient for fine patient screening--depending on the individual ocular parameters, it could be more restrictive. Advances in technology are needed to create a preoperative examination including two-dimensional maps of topography, pachymetry, and Young's modulus. PMID- 15796225 TI - Biomechanical modeling of corneal ectasia. PMID- 15796226 TI - Laser in situ keratomileusis for correction of myopia in eyes after retinal detachment surgery. AB - PURPOSE: To evaluate the safety and efficacy of laser in situ keratomileusis (LASIK) for correction of myopia in eyes with previous retinal detachment surgery. METHODS: A retrospective review was conducted of all consecutive eyes that underwent LASIK after retinal detachment surgery. Data was collected regarding previous ocular surgery and its time prior to LASIK, intra- and postoperative complications, and visual outcome measures pre- and postoperatively. RESULTS: Ten eyes with previous retinal detachment surgery were scheduled for LASIK. One eye was excluded from data analysis as LASIK was aborted due to inadequate microkeratome suction because of conjunctival scarring. Nine eyes underwent an uneventful LASIK procedure. On average, LASIK was performed 130+/-123 months following retinal detachment surgery. Postoperative LASIK follow up was 14.8+/-12.5 months. No significant intraoperative, postoperative, or retinal complications were observed. The mean preoperative spherical equivalent refraction was -9.00+/-3.00 diopters (D), uncorrected visual acuity (UCVA) was 0.06+/-0.02, and best spectacle-corrected visual acuity (BSCVA) was 0.64+/-0.16. At the end of follow-up, the mean spherical equivalent refraction was 0.65+/-0.88 D, mean UCVA was 0.57+/-0.14, and mean BSCVA was 0.72+/-0.19. Differences between BSCVA before and after LASIK were statistically significant (P=.038). At final follow-up, the safety index was 1.22 and efficacy index was 1.01. CONCLUSIONS: Laser in situ keratomileusis was found to be a safe and efficient option for treating refractive errors in eyes with previous retinal detachment surgery. PMID- 15796227 TI - Racial differences in central corneal thickness and refraction among refractive surgery candidates. AB - PURPOSE: To examine the preoperative central corneal thickness and refractive status of Caucasians, Hispanics, Asians, and African Americans in a refractive surgery patient population. METHODS: In a retrospective case series, a total of 803 consecutive eyes, which had no history of anterior segment surgery, underwent central corneal thickness and refraction measurement as part of a preoperative evaluation for refractive surgery from December 1999 to October 2002. Preoperative central corneal thickness was measured by a DGH-550 ultrasonic pachymeter, and spherical equivalence was determined by manifest refraction. RESULTS: This study included Caucasian (34.4%), Hispanic (34.5%), Asian (20.7%), African American (8.9%), and other (1.5%) patients. A one-way analysis of variance (ANOVA) revealed no significant differences in the central corneal thickness measurements among Asians, Hispanics, and Caucasians. However, mean central corneal thickness was 14 microm thinner in African Americans compared to Caucasians, and was significantly different at the 95% confidence interval. A one way ANOVA revealed a significant difference in the preoperative spherical equivalence among the four groups (F=4.91; df=3; P<.05) but no significant differences were noted in the preoperative astigmatism among any of the four groups (F=1.08; df=3; P>.05). A significant difference was noted in age in the four preoperative groups. CONCLUSIONS: Among the refractive population, African Americans were found to have thinner central corneal thickness than Caucasians. Asians were significantly (P<.05) more myopic compared to Caucasians, Hispanics, and African Americans. No difference was noted in the preoperative astigmatism among any of the four groups. Asians sought refractive surgery at a younger age whereas Caucasians sought it at an older age. PMID- 15796228 TI - LASIK with an inferior hinge. AB - PURPOSE: To investigate the refractive results and corneal pachymetric changes after laser in situ keratomileusis (LASIK) with an inferior hinge. METHODS: Forty six eyes of 23 patients who underwent LASIK with an inferior hinge were involved in this prospective study. All patients had a complete ophthalmic examination including uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), manifest refraction, computerized corneal topography (Orbscan II Corneal Topography System; Orbscan Inc, Salt Lake City, Utah), air-puff tonometry, slit-lamp examination of the anterior segment, and fundus examination. RESULTS: Preoperatively, mean sphere was -4.09+/-0.27 diopters (D) (range: -8.00 to -0.25 D), mean cylinder was -0.60+/-0.11 D (range: -3.25 to 0.00 D), and mean spherical equivalent refraction was -4.39+/-0.27 D (range: -8.63 to -1.50 D). At 1 month postoperatively, mean sphere was -0.04+/-0.01 D (range: -0.25 to 0.00 D), mean cylinder was -0.01+/-0.01 D (range: -0.50 to 0.00 D), and mean spherical equivalent refraction was -0.04+/-0.02 D (range: -0.00 to -0.50 D). At 1 month postoperatively, no eyes lost any lines of BSCVA. CONCLUSIONS: Laser in situ keratomileusis with an inferior hinge is an alternative technique, which was safe and effective in our series. Further studies with long-term follow-up comparing the incidence of corneal ectasia in eyes with a superior hinge to that in eyes with an inferior hinge are recommended. PMID- 15796229 TI - New formulas comparing the accommodation in human lens and intraocular lens. PMID- 15796231 TI - Holographic correction and phasing of large sparse-array telescopes. AB - I have constructed a 1-m-diameter telescope using separate, low-quality spherical primary mirror segments. A single hologram of the mirrors is used to correct the random surface distortions as well as spherical aberration, while simultaneously phasing the individual apertures together. I present experimental results of the removal of an error of thousands of waves to produce a diffraction-limited instrument operating over a narrow bandwidth. This technique promises to have many benefits in future space-based telescopes for imaging, lidar, and optical communications. PMID- 15796230 TI - Diurnal fluctuation of higher order ocular aberrations: correlation with intraocular pressure and corneal thickness. PMID- 15796232 TI - Analytic diffraction analysis of a 32-m telescope with hexagonal segments for high-contrast imaging. AB - Large segmented telescopes cannot be modeled accurately with fast-Fourier transform techniques since small features such as gaps between the segments will be inadequately sampled. An analytic Fourier-transform method can be used to model any pupil configuration with straight edges, including tolerance analysis and some types of apodization. We analytically investigated a 32-m segmented primary with 18 hexagonal segments for high-contrast imaging. There are significant regions in the image in which extrasolar planets could be detected. However, the hexagonal profile of the pupil was not as useful as expected. The gaps between the segments, the secondary obscuration, and the secondary spiders must be as small as possible and their edges must be apodized. Apodizing the edges of the individual segments reduced the useful regions in the image since the gaps appeared to be wider. PMID- 15796233 TI - Polarization characteristics of a wire-grid polarizer in a rotating platform. AB - I explore the polarization characteristics of a wire-grid polarizer operating in a rotating platform using well-established rigorous coupled-wave analysis. The results show that the performance of a wire-grid polarizer evaluated in transmittance and extinction ratio is substantially degraded with rotation, although it is still more robust than that of a perfect polarizer. The performance may further deteriorate with inclined incidence due to an increased negative effect of a Rayleigh anomaly. Either a finer or a deeper grating reduces the adverse effect of rotation. Although implementation of such a grating is difficult, the rotation characteristics can be enhanced by means of pixelating the wire grids of various orientations and capturing the input images with a sufficiently high sampling rate. PMID- 15796234 TI - Pattern recognition with generalized centroids and subcentroids. AB - We propose a class of generalized moment functions (GMFs) that can be used to determine a set of geometric points, namely, generalized centroids (G centroids), within an object. Based on a linear GMF, a mass centroid and its subcentroids can be defined and extracted, which provide information on the location and orientation of an object. Similar to traditional moment functions, GMFs can also be used to describe the global shape of an object, including symmetry and fullness. However, GMFs, along with G centroids and subcentroids, can further serve to construct a feature vector of an object, which is critical for image registration and invariant pattern recognition. One can extract more distinguishing features from the same object by changing the combination of GMFs. We present results that show simulations of pattern recognition from uniform backgrounds. PMID- 15796235 TI - Approximate method for the generalized M2 factor of rotationally symmetric hard edged diffracted flattened Gaussian beams. AB - On the basis of the truncated second-order moments method in the cylindrical coordinate systems and the expansion of the hard-edged aperture function into a finite sum of complex Gaussian functions, an approximate method used to calculate the generalized beam propagation factor (M2 factor) is proposed. The approximate analytical expressions of the generalized M2 factor for rotationally symmetric hard-edged diffracted flattened Gaussian beams defined by Gori [Opt. Commun. 107, 335 (1994)] and Li [Opt. Lett. 27, 1007 (2002)] are derived, respectively; we show that it depends on the beam order N and the beam truncation parameter delta. Some typical numerical examples are given to illustrate its applications that we compare by using the obtained analytical method and the numerical integration method. PMID- 15796236 TI - Full-field optical coherence tomography by achromatic phase shifting with a rotating polarizer. AB - We demonstrate two-dimensional detection optical coherence tomography (OCT) using achromatic phase shifting with a rotating polarizer. This phase shifting, which experiences a light beam with a cyclic change in its polarization state, is, in principle, independent of wavelength. We simulated the wavelength dependence of an achromatic phase shifter using Jones calculus and found that the achromatic region exceeded 145 nm when the deviation of the phase retardation was less than +/- 0.5 degrees. Using the achromatic phase shifter and a conventional phase shift calculation method, we obtained en face OCT images of an onion at different depths. This method is effective to enhance the quality of OCT with an ultrabroad spectrum light source. PMID- 15796237 TI - Grating projection system for surface contour measurement. AB - A grating projection system is a low-cost surface contour measurement technique that can be applied to a wide range of applications. There has been a resurgence of interest in the technique in recent years because of developments in computer hardware and image processing algorithms. We describe a method that projects a phase-shifted grating through a lens on an object surface. The deformed grating image on the object surface is captured by a CCD camera for subsequent analysis. Phase variation is achieved by a linear translation stage on which the grating is mounted. We compare the experimental results with the test results using a mechanical stylus method. PMID- 15796238 TI - Phase retrieval with a three-frame phase-shifting algorithm with an unknown phase shift. AB - A three-frame phase-shifting algorithm with a constant but unknown phase shift is proposed. The algorithm is based on background-intensity removal prior to phase retrieval to eliminate an undetermined factor in a fringe pattern. The proposed method is validated on three-dimensional profilometry by fringe projection and on deformation measurement by means of digital speckle shearing interferometry. For a fringe pattern with slow-varying background intensity, the background removal is achieved in the frequency domain. For a speckle pattern, a background removal technique is integrated with the three-frame algorithm. In this process, manual intervention is minimal, and high computational speed is achieved. In addition, high-frequency phase signals would not be removed in the noise-reduction process as is the case in the bandpass-filtering technique. Accuracy of the method is discussed. PMID- 15796239 TI - Two-dimensional thickness measurements based on internal reflection ellipsometry. AB - An imaging ellipsometer technique on internal reflection geometry that can measure the thickness distribution of a thin film possessing an assumed refractive index is described. Because a prism is used for the internal reflection geometry, it was theoretically predicted that angular derivation from the normal incidence on the prism surface affects only the psi value by a factor of 0.97 at maximum. Measurements were carried out for an optical system of silica substrate-TiO2 layer-silica layer-protein film-air, with a thin-film array of dried protein as the sample film. Thickness of the protein films was two dimensionally estimated only from the measured map of the delta value by use of the simulated relationship between the thickness and the delta value. The thickness map obtained was coincident on the whole with the results according to a mechanical scanning. The detection limit was approximately +/- 0.2 nm. These findings validate the optical effect of a high-index additional layer to improve the sensitivity and precision of thickness measurements of the sample film on transparent substrates. PMID- 15796240 TI - Properties of a diffused photon-pair density wave in a multiple-scattering medium. AB - A novel diffused photon-pair density wave (DPPDW) composed of correlated polarized photon pairs at different temporal frequencies and orthogonal linearly polarized states is proposed. A theory of DPPDWs is developed. A DPPDW selected by coherence gating and polarization gating that satisfies the diffusion equation has been verified experimentally. The sensitivity of amplitude and phase detection of the heterodyne signal has been improved by the properties of synchronized detection and common-path propagation of polarized pair photons in a multiple-scattering medium. Both reduced scattering coefficient micro2s' and absorption coefficient micro2alpha of the scattering medium in terms of the measured phase and amplitude of the heterodyne signal have been obtained. The detection sensitivity of micro2s' and micro2alpha and the properties of a DPPDW in a multiple-scattering medium are discussed and analyzed. PMID- 15796241 TI - Choroidal perfusion measurements made with optical coherence tomography. AB - Choroidal perfusion measurements are complicated by the choroid's location posterior to the retina and its associated retinal blood vessels. Optical coherence tomography is a relatively new imaging technique with sufficient spatial resolution to isolate choroidal backscattering events from the posterior eye. We modified a speckle imaging algorithm to analyze sequential axial depth scans obtained from posterior rat eye to obtain an indicator of choroidal perfusion. This indicator is correlated with known changes in choroidal blood flow in response to increased intraocular pressure. PMID- 15796242 TI - Experimental results and wear predictions of petal tools that freely rotate. AB - It is difficult to calculate the wear produced by free-pinned tools because their angular movement is not entirely predictable. We analyze the wear produced with free-pinned ring tools, using both simulations and experiments. We conclude that the wear of an incomplete ring is directly proportional to the ring's angular size, independently of the mean radius of the ring. We present an algorithm for calculation of the wear produced by free-pinned petal tools, as they can be considered a linear combination of incomplete free-pinned ring tools. Finally, we apply this result to the enhancement of a defective flat surface and to making a concave spheric surface. PMID- 15796243 TI - Asynchronous, all-optical signal processing based on the self-frequency shift of a gigahertz Raman soliton. AB - Ultrafast asynchronous all-optical signal processing is experimentally demonstrated. It is based on the intensity-dependent, self-frequency shift of a gigahertz Raman soliton. We demonstrate error-free, asynchronous, all-optical, bit-by-bit, self-signal recognition and demultiplexing from contended optical packets without use of an optical buffer, control pulse, or bit-phase synchronization. Fourfold, contended, 9.95-Gbit/s optical packets are transmitted through a conventional repeater span of 80 km and simultaneously demultiplexed to multiwavelength 9.95-Gbit/s optical packets with 0.5-dB processing sensitivity. Furthermore, we successfully accomplish demultiplexing from overlapping signals in contended optical packets with better than 3-dB recognition sensitivity. We confirm the capability of realizing a 3x cascade operation from bit-error-rate measurements. PMID- 15796244 TI - Clarifications to the paper "Holographic characteristics of a 1-mm-thick photopolymer to be used in holographic memories". AB - We have corrected typing errors related to the characterization of the dynamic range of the acrylamide photopolymer described in an earlier study [Appl. Opt. 42, 7008 (2003)]. The M number is expressed as M/# instead of M# as appears in the text. The value calculated from the experimental results that are included in the article is M/# = 3.8 instead of 38 as appears in the text. PMID- 15796245 TI - Theoretical and experimental studies of hologram multiplexing that uses a random wave front generated by photorefractive beam fanning. AB - A hologram multiplexing technique-that uses random wave fronts generated by photorefractive beam fanning is investigated. A storage photorefractive crystal generates various random wave fronts to be used as reference beams without the external diffusers such as ground glass and multimode optical fiber that are generally employed. We experimentally demonstrate hologram multiplexing with six images and show that the stored holograms can be selectively retrieved. We also simulate photorefractive beam fanning inside a BaTiO3 crystal, in particular regarding the correlation properties of the fanning beams for the first time to our knowledge, and reveal the conditions of incidence of an object beam and a reference beam required for suppressing image degradation, implementing low-cross talk retrieval, and producing a large number of stored holograms. PMID- 15796246 TI - Directional variation of visual range due to anisotropic atmospheric brightness. AB - The meteorological optical range (MOR) is the greatest distance at which a nonluminous object is barely discernible. Koschmieder related the optical atmospheric extinction coefficient to the contrast between the object and its background. His law assumes a uniform atmosphere and yields an isotropic value of the MOR. The model presented here incorporates the effect of anisotropic atmospheric brightness on contrast (and thus on the MOR). The Koschmieder MOR is then decreased by a factor according to the brightness prevailing in the direction of assessment. Manual assessments of the MOR made in arbitrary directions are shown to agree satisfactorily with the derated MOR. Minor modifications to existing instruments at airports would enable the instruments to register true values of the MOR and the runway visual range in directions relevant to the pilot, i.e., along the runway. PMID- 15796247 TI - Reflowed solgel spherical microlens for high-efficiency optical coupling between a laser diode and a single-mode fiber. AB - To improve the coupling efficiency between a laser diode and a single-mode fiber, we propose a two-microlens coupling scheme that uses two solgel spherical microlenses for high coupling efficiency. The conventional reflow technique was employed and extended to the inorganic-organic hybrid SiO2/ZrO2 solgel material to form the microlenses. Preliminary results show that the coupling efficiency was increased to--1.28 dB (74.5%) by the proposed scheme, compared with a coupling efficiency of--10.13 dB (9.7%) by the butt-joint method. The proposed fabrication technique demonstrates that use of a reflowed solgel spherical microlens is a cost-effective mass-production approach to application of micro optical elements in optical communication. PMID- 15796248 TI - Direct characterization of self-guided femtosecond laser filaments in air. AB - High-power femtosecond laser pulses propagating in air form self-guided filaments that can persist for many meters. Characterizing these filaments has always been challenging owing to their high intensity. An apparently novel diagnostic is used to directly measure the fluence distribution of femtosecond laser pulses after they have formed self-guided optical filaments in air. The diagnostic is unique in that the information contained in the filaments is not lost owing to the interaction with the apparatus. This allows filament characteristics such as energy and size to be unambiguously determined for the first time. PMID- 15796249 TI - Methods for optical adjustment in lidar systems. AB - In a lidar system, both a well-adjusted receiving telescope and an accurate transmitter-receiver alignment are essential for obtaining reliable data, but usually these are time consuming and difficult to accomplish, especially when the optical axis of the telescope is fixed. To solve these problems, we present a novel method, to our knowledge, for the receiving telescope adjustment, which is carried out mainly with a commercial laser plummet. A simple, fast alignment procedure is also described. These are tested by adjustment of the Fe-resonance fluorescence lidar system of Wuhan University and proved to be effective and convenient. The lidar system is applied to monitor the Fe layer in the mesosphere, leading to continuous retrieval of the iron density profiles with fine spatiotemporal resolution. PMID- 15796250 TI - Small-angle goniometry for backscattering measurements in the broadband spectrum. AB - We present experiments on spectral bidirectional reflectance distribution function (BRDF) effects at backscatter and discuss the feasibility of new methods for laboratory and field simulations of remote sensing of land surfaces. The extreme sharpness of the intensity peak allows both directional and comparative experimental spectral studies of hot spots. We demonstrate wavelength-dependent features in the hot-spot reflectance signatures that facilitate extension of spectral and directional BRDF measurements of natural targets (such as forest understories and ice surfaces) into retroreflection to exploit their hot-spot characteristics in the interpretation of spaceborne and airborne data. PMID- 15796251 TI - Combustion exhaust measurements of nitric oxide with an ultraviolet diode-laser based absorption sensor. AB - A diode-laser-based sensor has been developed for ultraviolet absorption measurements of the nitric oxide (NO) molecule. The sensor is based on the sum frequency mixing (SFM) of the output of a tunable, 395-nm external-cavity diode laser and a 532-nm diode-pumped, frequency-doubled Nd:YAG laser in a beta-barium borate crystal. The SFM process generates 325 +/- 75 nW of ultraviolet radiation at 226.8 nm, corresponding to the (v' = 0, v" = 0) band of the A2Sigma+-chi2II electronic transition of NO. Results from initial laboratory experiments in a gas cell are briefly discussed, followed by results from field demonstrations of the sensor for measurements in the exhaust streams of a gas turbine engine and a well stirred reactor. It is demonstrated that the sensor is capable of fully resolving the absorption spectrum and accurately measuring the NO concentration in actual combustion environments. Absorption is clearly visible in the gas turbine exhaust even for the lowest concentrations of 9 parts per million (ppm) for idle conditions and for a path length of 0.51 m. The sensitivity of the current system is estimated at 0.23%, which corresponds to a detection limit of 0.8 ppm in 1 m for 1000 K gas. The estimated uncertainty in the absolute concentrations that we obtained using the sensor is 10%. PMID- 15796252 TI - Wide-field-of-view GaAs/AlxOy one-dimensional photonic crystal filter. AB - The design, fabrication, and characterization of a one-dimensional photonic crystal optical filter that has a relatively narrow, flat-topped passband within a wide stop band and small angular sensitivity is presented. The filter is based on a one-dimensional photonic crystal structure that has multiple defects, facilitating simultaneous minimization of the angular sensitivity and optimization of the passband's characteristics. We use epitaxially grown and selectively oxidized GaAs/AlxOy multilayers to achieve a high-index-contrast material system and incorporate the experimentally determined optical and material properties into the design of the device. A flat-topped bandpass filter with a bandwidth of 65 nm and a wide field of view of 50 degrees is experimentally characterized and compared with the design predictions. PMID- 15796253 TI - Surgical management of strabismus for dysthyroid ophthalmopathy. AB - BACKGROUND: The study reports our 8-year experience in the surgical treatment of restrictive myopathy resulting from dysthyroid ophthalmopathy. We tried to determine the factors that contributed to favorable surgical outcomes. METHODS: The charts of patients who had surgical treatment for strabismus related to dysthyroid ophthalmopathy at Chang Gung Memorial Hospital between January 1995 and December 2002 (n=25) were retrospectively reviewed, and factors that possibly influenced the outcome were statistically analyzed for significance. RESULTS: The mean pre-operative vertical deviation of the 25 patients was 23.3 prism diopters and the pre-operative horizontal deviation was 11.3 prism diopters. The mean follow-up time was 29.7 months. Each patient had strabismus surgery an average of 1.5 times. The majority of patients (84%) were satisfied with the result. No significant difference in the success rate was evident between patients who waited less than 3 months before the surgery and those who waited for a longer period. The repeat surgery rate was significantly higher in the latter group. A shorter duration of diplopia and smaller angle of pre-operative horizontal deviation were contributors towards a favorable outcome. CONCLUSIONS: A waiting time of less than 3 months prior to treatment of strabismus associated with dysthyroid ophthalmopathy produced a favorable outcome. The duration of diplopia and the pre-operative angle of horizontal deviation were the most important prognostic factors. PMID- 15796255 TI - Risk factors for first-ever ischemic stroke: a hospital-based case-control study in Kaohsiung, Taiwan. AB - BACKGROUND: Numerous population-based epidemiological studies have shown the prevalence and strength of vascular risk factors for stroke but little is known about risk factors of stroke in Taiwan. The aim of this study was to identify vascular risk factors in a group of first-ever ischemic stroke patients in Taiwan. METHODS: The study consisted of 228 consecutively hospitalized cases of first-ever ischemic stroke and 228 healthy age- and sex-matched control subjects. Conditional logistic regression analyses were performed to evaluate the risk factors. RESULTS: Significant risk factors included hypertension (odds ratio [OR] 2.7, 95% confidence interval [CI] = 1.53-4.80), atrial fibrillation (OR 14.8, 95% CI = 2.32-94.73), ischemic heart disease (OR 4.4, 95% CI = 1.48-13.38), cigarette smoking (OR 2.3, 95% CI = 1.10-4.96), left ventricular hypertrophy (OR 2.7, 95% CI = 1.18-6.16), and other abnormal electrocardiographic findings (OR 2.1, 95% CI = 1.11-3.80). CONCLUSIONS: Several vascular risk factors of first-ever ischemic stroke were identified. A population-based study involving more vascular risk factors is needed for generalization. PMID- 15796254 TI - Plasma endothelin-1 levels in patients with rheumatic mitral stenosis and a history of cerebral thromboembolism. AB - BACKGROUND: Increased plasma endothelin-1 concentrations have been observed in patients with rheumatic mitral stenosis. Endothelin-1 levels have never been investigated in patients with mitral stenosis and history of cerebral thromboembolism. METHODS: We measured plasma concentrations of endothelin-1 in the peripheral venous blood samples obtained from 20 patients with moderate to severe rheumatic mitral stenosis (16 with permanent atrial fibrillation and 4 with sinus rhythm). Six patients had history of thromboembolism. The remaining 14 patients did not have history of thromboembolism. Plasma endothelin-1 concentrations were measured using solid phase sandwich enzyme linked-immuno sorbent assay. RESULTS: The peripheral venous concentrations of endothelin-1 of the six patients with history of thromboembolism did not differ from the concentrations of the 14 patients without history of thromboembolism (2.40 +/- 1.39 pg/ml vs. 2.49 +/- 0.66 pg/ml, p = 0.9). CONCLUSIONS: Although plasma endothelin-1 concentrations were increased in patients with mitral stenosis, plasma endothelin-1 concentrations were not further elevated in patients with mitral stenosis and history of thromboembolism. PMID- 15796256 TI - Magnetic resonance imaging characteristics of benign and malignant vertebral fractures. AB - BACKGROUND: Attempts to differentiate benign and malignant vertebral fractures may be difficult, particularly when there is no obvious evidence of malignancy. Since early diagnosis and appropriate management of malignant vertebral fractures are important, a reliable imaging modality is required. METHODS: From January 1996 to December 2002, 48 patients with malignant vertebral fractures and 50 patients with benign processes were studied. All patients underwent conventional magnetic resonance imaging (MRI) scanning for acute vertebral compression fractures within 2 months of presenting with the complaint. Seven MRI characteristics were used as criteria, including signal intensity, gadolinium enhancement, epidural compression, multiple compression fractures, associated paraspinal soft tissue mass, pedicle involvement, and posterior element involvement. The predictive value of each MRI characteristic for distinguishing malignant from benign osteoporotic vertebral fractures was tested by statistical analysis. RESULTS: Lesions with negative gadolinium enhancement were favored as benign fractures. A uniform signal change in multiple involved vertebra lesions, round, smooth margins with marked epidural compression, a paraspinal soft tissue mass, and pedicle and posterior element involvement were probable malignant characteristics. Among them an associated paraspinal soft tissue mass was found to be significant in predicting the probability of malignancy. CONCLUSIONS: Certain MRI characteristics allow early differentiation of benign and malignant vertebral fractures. PMID- 15796257 TI - Brachiobasilic fistula as a secondary access procedure: an alternative to a dialysis prosthetic graft. AB - BACKGROUND: In order to extend the availability of each extremity as an access site for long-term dialysis, we created a brachiobasilic fistula by superficialization of the basilic vein, which had previously been arterialized. In this work, we present the preliminary results of this procedure. METHODS: Patients with prior forearm arteriovenous fistula dysfunction without an adequate superficial vein were enrolled in this study. The basilic vein was superficialized to create a brachiobasilic fistula. The basilic veins were transposed in 16 patients and elevated in 4 patients. At the same time, another 50 upper arm bridge prosthetic grafts were created. Finally, surgery-related complications and patency rates were analyzed and compared with upper arm prosthetic grafts. RESULTS: Twenty brachiobasilic fistulas and 50 upper arm bridge prosthetic grafts were created in 70 patients in our hospital from September 2001 to January 2004. No surgery-related deaths occurred. Cannulation of the brachiobasilic fistulas began at an average of 33 (range, 21 to 84) days after surgery. All fistulas were successfully used for dialysis. The mean follow up was 15 (range, 2 to 26) months. Moreover, the primary patency rates for the brachiobasilic fistulas and upper arm prosthetic grafts were 80% and 70%, respectively. Notably, all brachiobasilic fistulas remained patent at the last follow-up. Furthermore, surgery-related complications were 25% for brachiobasilic fistulas and 60% for upper arm prosthetic grafts. Common complications of brachiobasilic fistulas were graft thrombosis, arm swelling, hematoma formation, and steal syndrome. CONCLUSIONS: Brachiobasilic fistulas are technically feasible and show excellent patency. Fewer surgery-related complications were seen compared to upper arm prosthetic grafts, and most of them were treated without loss of the fistula. We suggest that this procedure should be considered before placement of a prosthetic graft whenever feasible. These observations merely represent our preliminary experience regarding this procedure, and further investigation involving larger cases numbers is needed in the future. PMID- 15796258 TI - Anemia as the sole presenting symptom of idiopathic pulmonary hemosiderosis: report of two cases. AB - Idiopathic pulmonary hemosiderosis (IPH) is a rare disease in children and has an unknown etiology. It is characterized by the triad of hemoptysis, pulmonary infiltrates on chest radiograph (CXR) and iron deficiency anemia. We report two young children, aged 3 and 4 years, were admitted due to pale-looking appearance but without hemoptysis or other respiratory symptoms. Pallor was the sole presenting feature in these 2 children with IPH and which was unusual. CXR obtained on admission led to the suspicion of pulmonary hemorrhage. The diagnosis of IPH was confirmed based on the presence of many hemosiderin-laden macrophages in bronchoalveolar lavage fluid obtained by flexible bronchoscopy. Steroid was initiated after the diagnosis of IPH was established; the both of them have been well and received regular follow-up in our outpatient department. IPH may not be diagnosed because of difficulty in diagnosis. Anemia may be the only presenting feature of IPH, which was due to occult pulmonary hemorrhage. Initial treatment with corticosteroids has been successful in our patients for a period of 6 and 8 months of follow up respectively. PMID- 15796259 TI - Postoperative endophthalmitis by Flavimonas oryzihabitans. AB - A 55-year-old man suffered from blurred vision of the left eye (visual acuity was counting fingers/5-10 cm) 8 days after undergoing cataract surgery. After thorough ophthalmologic examinations with gram-negative bacillus justified on smear of vitreous specimen, postoperative endophthalmitis (os) was impressed. Intravitreous injection of 1.0 mg of vancomycin, 0.4 mg of amikacin, and 0.4 mg of dexamethasone was given twice and vitreous fluid culture was also performed. However, the inflammatory condition was not well controlled. Because the results of vitreous fluid culture revealed Flavimonas oryzihabitans infection which is sensitive to piperacillin, we performed the 3rd intravitreous injection of 1.0 mg of piperacillin and 0.4 mg of dexamethasone. The inflammatory condition was under control and the visual acuity of left eye improved. For patients with ocular diseases, postoperative endophthalmitis by Flavimonas oryzihabitans is a rare condition. According to the results of this case, intravitreous injection of piperacillin was effective against the pathogen. PMID- 15796260 TI - Chronic hepatitis C virus infection associated with dermatomyositis and hepatocellular carcinoma. AB - Dermatomyositis is a rare and idiopathic inflammatory myopathy with characteristic cutaneous manifestations. In recent years, some researchers have showed the cause of dermatomyositis might be due to an autoimmune response induced by viral infections. However, chronic hepatitis C virus (HCV) infection associated with dermatomyositis is very rare. In this report, we present a patient with dematomyositis with abnormal liver function test results and elevated alfa-fetoprotein level. After excluding multiple viral infections known to cause myositis, the case was proven to be chronic hepatitis C by positive HCV RNA in the serum. Abdominal computed tomography showed a liver tumor on the right lobe and needle biopsy proved it to be hepatocellular carcinoma. Chronic hepatitis C or hepatocellular carcinoma might cause dermatomyositis by inducing the formation of autoantibodies. Chronic hepatitis C or hepatocellular carcinoma should be considered in patients of dermatomyositis if no other cause is found. PMID- 15796261 TI - Presentation of prostate carcinoma with cervical lymphadenopathy: report of three cases. AB - The most common method of spreading of prostate cancer is via direct invasion to the pelvic organs or vertebral bodies. Supraclavicular lymph node metastasis is uncommon in patients with prostate cancer. Previously researchers have shown that the prognosis after such a presentation is generally poor. We herein present three patients with prostate carcinoma presenting with supraclavicular lymph node enlargement at the initial diagnosis. Abdominal computed tomography showed multiple enlarged lymph nodes in retroperitoneal space. All of our patients had hormone therapy and the prostate specific antigen (PSA) level declined. The mean follow up time was 16 months (range, 6-24 months). None of our patients experienced progression during follow up. Prostate carcinoma should always be considered in the differential diagnosis of elderly men with cervical lymphadenopathy, even in the absence of lower urinary tract symptoms. Evaluation of the PSA level and lymph node biopsy with PSA immunohistochemistry stain can lead to the final diagnosis. In addition, hormone treatment has been shown to be of benefit even in patients in the advanced stages. PMID- 15796262 TI - Gelatinous degeneration of the bone marrow in anorexia nervosa. AB - Anorexia nervosa is a chronic psychiatric process characterized by a restrictive disorder in alimentary habits. Hematologic alterations in the peripheral blood include cytopenias involving one or more hematopoietic lineages. Morphologic changes in the bone marrow and stereologic alterations in bone marrow adiopocytes may also be observed in anorexia nervosa. We present a 12-year-old girl who had chronic anorexia and one third of body weight loss during an 8-month period. She was apathetic and had missed several menstrual cycles. The sex maturity rating was Tanner stage IV. There was no lymphadenopathy, no hepatosplenomegaly, and no identifiable tumor mass. She was not anemic, but was found to have leukopenia, neutropenia and a low level of triiodothyronine. Sections of the bone marrow biopsy showed almost complete serous atrophy (gelatinous degeneration) of the bone marrow. In this patient, the bone marrow alteration is related to nutritional deprivation of anorexia nervosa. PMID- 15796263 TI - Disruptive doc poses risk to group. PMID- 15796264 TI - Product certification process under way to assist medical groups. PMID- 15796265 TI - A picture is worth 1,000 words. PMID- 15796266 TI - Finders and keepers: locating and retaining the right staff. PMID- 15796267 TI - Giddyup. PMID- 15796268 TI - The big benefit squeeze: companies are paring costs by clamping down on healthcare. PMID- 15796269 TI - Mind movies. PMID- 15796270 TI - Laughter: a hearty har-har. PMID- 15796271 TI - To create, or not to create? PMID- 15796272 TI - Double, double, toil and trouble: infections still spreading in long-term-care facilities. PMID- 15796273 TI - Standardized infection surveillance in long-term care: interfacility comparisons from a regional cohort of facilities. AB - OBJECTIVES: To measure infection rates in a regional cohort of long-term-care facilities (LTCFs) using standard surveillance methods and to analyze different methods for interfacility comparisons. SETTING: Seventeen LTCFs in Idaho. DESIGN: Prospective, active surveillance for LTCF-acquired infections using standard definitions and case-finding methods was conducted from July 2001 to June 2002. All surveillance data were combined and individual facility performance was compared with the aggregate employing a variety of statistical and graphic methods. RESULTS: The surveillance data set consisted of 472,019 resident-days of care with 1717 total infections for a pooled mean rate of 3.64 infections per 1000 resident-days. Specific infections included respiratory (828; rate, 1.75), skin and soft tissue (520; rate, 1.10), urinary tract (282; rate, 0.60), gastrointestinal (77; rate, 0.16), unexplained febrile illnesses (6; rate, 0.01), and bloodstream (4; rate, 0.01). Initially, methods adopted from the National Nosocomial Infections Surveillance System were used comparing individual rates with pooled means and percentiles of distribution. A more sensitive method appeared to be detecting statistically significant deviations (based on chi square analysis) of the individual facility rates from the aggregate of all other facilities. One promising method employed statistical process control charts (U charts) adjusted to compare individual rates with aggregate monthly rates, providing simultaneous visual and statistical comparisons. Small multiples graphs were useful in providing images valid for rapid concurrent comparison of all facilities. CONCLUSION: Interfacility comparisons have been demonstrated to be valuable for hospital infection control programs, but have not been studied extensively in LTCFs. PMID- 15796274 TI - Persistence of fluoroquinolone-resistant, multidrug-resistant Streptococcus pneumoniae in a long-term-care facility: efforts to reduce intrafacility transmission. AB - OBJECTIVE: We describe an effort to reduce transmission of a multidrug-resistant Streptococcus pneumoniae (MDRSP) in a long-term-care facility (LTCF). DESIGN: Longitudinal cross-sectional study. SETTING: An LTCF in New York City with ongoing disease due to an MDRSP strain among residents with AIDS since a 1995 outbreak. The MDRSP outbreak strain was susceptible to vancomycin but not to other antimicrobials tested, including fluoroquinolones. PARTICIPANTS: Residents and staff members of the LTCF during 1999 through 2001. INTERVENTION: Implementing standard infection control measures, and developing and implementing "enhanced standard" infection control measures, modified respiratory droplet prevention measures to reduce inter-resident transmission. RESULTS: Before the intervention, nasopharyngeal carriage of the MDRSP outbreak strain was detected in residents with AIDS and residents with tracheostomies who were not dependent on mechanical ventilation. The prevalence of nasopharyngeal carriage of the MDRSP outbreak strain was 7.8% among residents who had AIDS and 14.6% among residents with tracheostomies. After training sessions on standard and enhanced standard infection control measures, the staff appeared to have good knowledge and practice of the infection control measures. After the intervention, new transmission among residents with tracheostomies was prevented; however, these residents were prone to persistent tracheal carriage and needed ongoing enhanced standard infection control measures. Ongoing transmission among residents with AIDS, a socially active group, was documented, although fewer cases of disease due to the outbreak strain occurred. CONCLUSIONS: Infection control contributed to less transmission of MDRSP in the LTCE Additional strategies are needed to reduce transmission and carriage among certain resident populations. PMID- 15796276 TI - A norovirus gastroenteritis epidemic in a long-term-care facility. AB - BACKGROUND: In Victoria, Australia, from July to December 2002, 126 outbreaks of viral and suspected viral gastroenteritis were reported in healthcare institutions. Norovirus was found to account for at least 77 of the 126 outbreaks. METHODS: In October 2002, the infection control unit investigated an outbreak of acute gastroenteritis on three wards in a 500-bed, long-term-care facility in Melbourne, Victoria, Australia. Cohorting and other infection control measures were initiated. RESULTS: The outbreak was controlled 32 days after the first symptoms of acute gastroenteritis were identified. Fifty-two patients and 11 staff members were affected. Norovirus genotype 2 was detected on two of the three wards. Norovirus was not isolated in the third ward but was suspected to be the causative organism. CONCLUSIONS: Outbreaks of viral gastroenteritis can cause significant morbidity in a long-term-care facility, affecting both patients and staff. In addition, the transmission of viral pathogens can be well established before there is recognition of an outbreak. PMID- 15796275 TI - Failure to control an outbreak of multidrug-resistant Streptococcus pneumoniae in a long-term-care facility: emergence and ongoing transmission of a fluoroquinolone-resistant strain. AB - OBJECTIVES: To characterize risk factors associated with pneumococcal disease and asymptomatic colonization during an outbreak of multidrug-resistant Streptococcus pneumoniae (MDRSP) among AIDS patients in a long-term-care facility (LTCF), evaluate the efficacy of antimicrobial prophylaxis in eliminating MDRSP colonization, and describe the emergence of fluoroquinolone resistance in the MDRSP outbreak strain. DESIGN: Epidemiologic investigation based on chart review and characterization of SP strains by antimicrobial susceptibility testing and PFGE and prospective MDRSP surveillance. SETTING: An 80-bed AIDS-care unit in an LTCF PARTICIPANTS: Staff and residents on the unit. RESULTS: From April 1995 through January 1996, 7 cases of MDRSP occurred. A nasopharyngeal (NP) swab survey of all residents (n=65) and staff (n=70) detected asymptomatic colonization among 6 residents (9%), but no staff. Isolates were sensitive only to rifampin, ofloxacin, and vancomycin. A 7-day course of rifampin and ofloxacin was given to eliminate colonization among residents: NP swab surveys at 1, 4, and 10 weeks after prophylaxis identified 1 or more colonized residents at each follow-up with isolates showing resistance to one or both treatment drugs. Between 1996 and 1999, an additional 6 patients were diagnosed with fluoroquinolone-resistant (FQ-R) MDRSP infection, with PFGE results demonstrating that the outbreak strain had persisted 3 years after the initial outbreak was recognized. CONCLUSIONS: Chemoprophylaxis likely contributed to the development of a FQ-R outbreak strain that continued to be transmitted in the facility through 1999. Long-term control of future MDRSP outbreaks should rely primarily on vaccination and strict infection control measures. PMID- 15796277 TI - An outbreak of norovirus infection in a long-term-care unit in Spain. AB - BACKGROUND: Norovirus belongs to the Caliciviridae family and causes outbreaks of infectious enteritis by fecal-oral transmission. In Spain, there have been few outbreaks reported due to this virus. We describe an outbreak on a long-term-care hospital ward. METHODS: Cases were classified as probable, confirmed, and secondary. Stool cultures were performed. Polymerase chain reaction detection of norovirus was also performed. RESULTS: The outbreak occurred from December 7 to 28, 2001, involving 60 cases (32 patients, 19 staff members, 8 patients' relatives, and 1 relative of a staff member). Most (82%) of the cases were female. The most frequently involved ages were 20 to 39 years for staff members and 70 to 89 years for patients. The incubation period of secondary cases in patients' families had a median of 48 hours (range, 1 to 7 days). Clinical symptoms included diarrhea (85%), vomiting (75%), fever (37%), nausea (23%), and abdominal pain (12%). Median duration of the disease was 48 hours (range, 1 to 7 days). All cases resolved and the outbreak halted with additional hygienic measures. Stool cultures were all negative for enteropathogenic bacteria and rotaviruses. In 16 of 23 cases, the norovirus genotype 2 antigen was detected. CONCLUSION: This outbreak of gastroenteritis due to norovirus genotype 2 affected patients, staff members, and their relatives in a long-term-care facility and was controlled in 21 days. PMID- 15796278 TI - Impact of an outbreak of norovirus infection on hospital resources. AB - OBJECTIVE: To describe a nosocomial norovirus outbreak, its management, and its financial impact on hospital resources. DESIGN: A matched case-control study and microbiological investigation. METHODS: We compared 16 patients with norovirus infection with control-patients matched by age, gender, disease category, and length of stay. Bed occupancy-days during the peak incidence period of the outbreak were compared with the corresponding periods in 2001 and 2002. Expenses due to increased workload were calculated based on a measuring system that records time spent for nursing care per patient per day. RESULTS: The attack rates were 13.9% among patients and 29.5% among healthcare workers. The median number of occupied beds was significantly lower due to bed closure during the peak incidence in 2003 (29) compared with the median number of occupied beds in 2001 and 2002 combined (42.5). Based on this difference and a daily charge of 562.50 dollars per patient, we calculated a revenue loss of 37,968 dollars. Additional expenses totaled 10,300 dollars for increased nursing care. Extra costs for microbiological diagnosis totaled 2707 dollars. Lost productivity costs due to healthcare workers on sick leave totaled 12,807 dollars. The expenses for work by the infection control team totaled 1408 dollars. The financial impact of this outbreak on hospital resources comprising loss of revenue and extra costs for microbiological diagnosis but without lost productivity costs, increased nursing care, and expenses for the infection control team totaled 40,675 dollars. CONCLUSIONS: Nosocomial norovirus outbreaks result in significant loss of revenue and increased use of resources. Bed closures had a greater impact on hospital resources than increased need for nursing care PMID- 15796279 TI - Evaluation of the impact of the source (patient versus staff) on nosocomial norovirus outbreak severity. AB - OBJECTIVE: To study the dependence of infection risk and outbreak size on the type of index case (i.e., patient or staff). METHODS: Nosocomial outbreaks were reviewed and categorized into those started by patients and those started by staff. Infection risks and outbreak sizes were evaluated taking into account the index case category. RESULTS: Of the 30 nosocomial outbreaks of norovirus with person-to-person transmission, 20 (67%) involved patients as the index cases. Patient-indexed outbreaks affected significantly more patients than did staff indexed outbreaks (difference in means, 16.25; 95% confidence interval [CI95], 5.1 to 27.0). For the numbers of affected staff, no dependence on the index case category was detectable (difference in means, -1.05; CI95, -9.0 to 6.9). For patients exposed during patient-indexed outbreaks, the risk of acquiring a norovirus infection was approximately 4.8 times as high as the corresponding risk for patients exposed during staff-indexed outbreaks (odds ratio [OR], 4.79; CI95, 1.82 to 8.28). The infection risk for exposed staff during patient-indexed outbreaks was approximately 1.5 times as high as the corresponding risk during staff-indexed outbreaks (OR, 1.51; CI95, 0.92 to 2.49). CONCLUSIONS: Patient indexed norovirus outbreaks generally affect more patients than do staff-indexed outbreaks. Staff appear to be similarly affected by both outbreak index category groups. This study demonstrates the importance of obtaining complete outbreak data, including the index case classification as staff or patient, during norovirus outbreak investigations. Such information may be useful for further targeting prevention measures PMID- 15796281 TI - Impact of nosocomial infection on cost of illness and length of stay in intensive care units. AB - OBJECTIVE: Economic evaluation has become increasingly important in healthcare and infection control. This study evaluated the impact of nosocomial infections on cost of illness and length of stay (LOS) in intensive care units (ICUs). DESIGN: A retrospective cohort study. SETTING: Medical, surgical, and mixed medical and surgical ICUs in a tertiary-care referral medical center. PATIENTS: Patients admitted to adult ICUs between October 2001 and June 2002 were eligible for the study. METHODS: Estimates of the cost and LOS for patients who acquired a nosocomial infection were computed using a stratified analysis and regression approach. RESULTS: During the study period, 778 patients were admitted to the ICUs. Total costs for patients with and without nosocomial infections (median cost, 10,354 dollars and 3985 dollars, respectively) were significantly different (P < .05). The costs stratified by infection site (median differences from 4687 dollars to 7365 dollars) and primary diagnosis (median differences from 5585 dollars to 16,507 dollars) were also significantly different (P < .05) except for surgical-site infection. After covariates were adjusted for in the multiple linear regression, nosocomial infection increased the total costs by 3306 dollars per patient and increased the LOS by 18.2 days per patient (P < .001). Each additional day spent in the ICU increased the cost per patient by 353 dollars (P < .001). CONCLUSIONS: Nosocomial infections are associated with increased cost of illness and LOS. Prevention of nosocomial infections should reduce direct costs and decrease the LOS. PMID- 15796280 TI - A large outbreak of Clostridium difficile-associated disease with an unexpected proportion of deaths and colectomies at a teaching hospital following increased fluoroquinolone use. AB - BACKGROUND AND OBJECTIVE: Fluoroquinolones have not been frequently implicated as a cause of Clostridium difficile outbreaks. Nosocomial C. difficile infections increased from 2.7 to 6.8 cases per 1000 discharges (P < .001). During the first 2 years of the outbreak, there were 253 nosocomial C. difficile infections; of these, 26 resulted in colectomy and 18 resulted in death. We conducted an investigation of a large C. difficile outbreak in our hospital to identify risk factors and characterize the outbreak. METHODS: A retrospective case-control study of case-patients with C. difficile infection from January 2000 through April 2001 and control-patients matched by date of hospital admission, type of medical service, and length of stay; an analysis of inpatient antibiotic use; and antibiotic susceptibility testing and molecular subtyping of isolates were performed. RESULTS: On logistic regression analysis, clindamycin (odds ratio [OR], 4.8; 95% confidence interval [CI95], 1.9-12.0), ceftriaxone (OR, 5.4; CI95, 1.8-15.8), and levofloxacin (OR, 2.0; CI95, 1.2-3.3) were independently associated with infection. The etiologic fractions for these three agents were 10.0%, 6.7%, and 30.8%, respectively. Fluoroquinolone use increased before the onset of the outbreak (P < .001); 59% of case-patients and 41% of control patients had received this antibiotic class. The outbreak was polyclonal, although 52% of isolates belonged to two highly related molecular subtypes. CONCLUSIONS: Exposure to levofloxacin was an independent risk factor for C. difficile-associated diarrhea and appeared to contribute substantially to the outbreak. Restricted use of levofloxacin and the other implicated antibiotics may be required to control the outbreak PMID- 15796282 TI - Health and economic consequences of an outbreak of pertussis among healthcare workers in a hospital in France. AB - BACKGROUND: Bordetella pertussis is highly contagious, and because immunity wanes after vaccination, it continues to be a cause of cough among adults. OBJECTIVE: To describe the healthcare services used and productivity losses accrued by healthcare workers (HCWs) missing work due to pertussis. METHODS: After 3 pertussis cases were confirmed among HCWs, all hospital employees and patients with a cough were screened between November 2000 and March 2001. Each potential case underwent diagnostic tests and received antibiotics (spiramycin or azithromycin) when appropriate. Symptomatic employees were not allowed to return to work until they received an antibiotic for at least 5 days. Services used (physician visits and calls, antibiotics, diagnostic tests, hospitalization, and treatment provided to their contacts) were combined with cost estimates (in 2002 euros) for these services in France. RESULTS: Ninety-one potential cases were identified (77 HCWs, 12 patients, and 2 family members). Of them, 89% received antibiotics and 22% had at least one contact who was also treated. Approximately half (55%) of the HCWs who were cases missed 5 days of work. Four patients were admitted to the hospital as a result of the infection. The average medical cost was 297 euros per potential case: diagnostic tests accounted for 32% and hospitalization for 31%. Total cost (medical and productivity) was 46,661 euros for 91 cases, 42% from productivity losses. An investigation to identify these potential cases also accrued additional costs. CONCLUSION: Serious adverse health and economic consequences arose from transmission of pertussis among HCWs, their families, and patients. PMID- 15796283 TI - Effect of pressure ulcers on length of hospital stay. AB - OBJECTIVE: To identify the independent effect of pressure ulcers on excess length of stay and control for all observable factors that may also contribute to excess length of stay. Hospitalized patients who develop a pressure ulcer during their hospital stay are at a greater risk for increased length of stay as compared with patients who do not. DESIGN: Cross-sectional, observational study. SETTING: Tertiary-care referral and teaching hospital in Australia. PATIENTS: Two thousand hospitalized patients 18 years and older who had a minimum stay in the hospital of 1 night and admission to selected clinical units. METHODS: Two thousand participants were randomly selected from 4,500 patients enrolled in a prospective survey conducted between October 2002 and January 2003. Quantile median robust regression was used to assess risk factors for excess length of hospital stay. RESULTS: Having a pressure ulcer resulted in a median excess length of stay of 4.31 days. Twenty other variables were statistically significant at the 5% level in the final model. CONCLUSIONS: Pressure ulcers make a significant independent contribution to excess length of hospitalization beyond what might be expected based on admission diagnosis. However, our estimates were substantially lower than those currently used to make predictions of the economic costs of pressure ulcers; existing estimates may overstate the true economic cost. PMID- 15796284 TI - Knowledge of standard and isolation precautions in a large teaching hospital. AB - OBJECTIVE: To assess the level of knowledge regarding and attitudes toward standard and isolation precautions among healthcare workers in a hospital. METHOD: A confidential, self-administered questionnaire survey was conducted in a random sample of 1500 nurses and 500 physicians in a large teaching hospital. RESULTS: A total of 1,241 questionnaires were returned (response rate, 62%). The median age of respondents was 39 years; 71.9% were women and 21.2% had senior staff status. One-fourth had previously participated in specific training regarding transmission precautions for pathogens conducted by the infection control team. More than half (55.9%) gave correct answers to 10 or more of the 13 knowledge-type questions. The following reasons for noncompliance with guidelines were judged as "very important": lack of knowledge (47%); lack of time (42%); forgetfulness (39%); and lack of means (28%). For physicians and healthcare workers in a senior position, lack of time and lack of means were significantly less important (P < .0005). On multivariate linear regression, knowledge was independently associated with exposure to training sessions (coefficient, 0.33; 95% confidence interval, 0.08 to 0.57; P = .009) and less professional experience (coefficient per increasing professional experience, -0.024; 95% confidence interval, -0.035 to -0.012; P < .0005). CONCLUSIONS: Despite a training effort targeting opinion leaders, knowledge of transmission precautions for pathogens remained insufficient. Nevertheless, specific training proved to be the major determinant of "good knowledge". PMID- 15796285 TI - Attitudes and perceptions toward hand hygiene among healthcare workers caring for critically ill neonates. AB - BACKGROUND: Infectious complications are frequent among critically ill neonates. Hand hygiene is the leading measure to prevent healthcare-associated infections, but poor compliance has been repeatedly documented, including in the neonatal setting. Hand hygiene promotion requires a complex approach that should consider personal factors affecting healthcare workers' attitudes. OBJECTIVE: To identify beliefs and perceptions associated with intention to comply with hand hygiene among neonatal healthcare workers. METHODS: An anonymous, self-administered questionnaire (74 items) based on the theory of planned behavior was distributed to 80 neonatal healthcare workers to assess intention to comply, attitude toward hand hygiene, behavioral and subjective norm perceptions, and perception of difficulty to comply. Variables were assessed using multi-item measures and answers to 7-point bipolar scales. All multi-item scales had satisfactory internal consistency (alpha > 0.7). Multivariate logistic regression identified independent perceptions or beliefs associated with a positive intention to comply. RESULTS: The response rate was 76% (61 of 80). Of the 49 nurses and 12 physicians responding, 75% believed that they could improve their compliance with hand hygiene. Intention to comply was associated with perceived control over the difficulty to perform hand hygiene (OR, 3.12; CI95, 1.12 to 8.70; P = .030) and a positive perception of how superiors valued hand hygiene (OR, 2.89; CI95, 1.08 to 7.77; P = .035). CONCLUSION: Our data highlight the importance of the opinions of superiors and a strong perceived controllability over the difficulty to perform hand hygiene as possible internal factors that may influence hand hygiene compliance. PMID- 15796286 TI - Compliance with hand hygiene and glove use in a university-affiliated hospital. AB - OBJECTIVE: The hands of healthcare workers often transmit pathogens causing nosocomial infections. This study examined compliance with handwashing and glove use. SETTING: A university-affiliated hospital. DESIGN: Compliance was observed covertly. Healthcare workers' demographics, hand hygiene facilities, indications for hand hygiene, compliance with handwashing and glove use in each procedure, and duration of handwashing were recorded. RESULTS: Nine nurses and 33 assistant physicians were monitored during the study. One researcher recorded 1400 potential opportunities for handwashing during 15-minute observation periods. The mean duration of handwashing was 10 +/- 2 seconds. Most healthcare workers (99.3%) used liquid soap during handwashing, but 79.8% did not dry their hands. For all indications, compliance with handwashing was 31.9% and compliance with glove use was 58.8%. Compliance with handwashing varied inversely with both the number of indications for hand hygiene and the number of patient beds in the hospital room. Compliance with handwashing was better in dirty high-risk situations. CONCLUSION: Compliance with handwashing was low, suggesting the need for new motivational strategies such as supplying feedback regarding compliance rates PMID- 15796287 TI - Efficiency of hand drying for removing bacteria from washed hands: comparison of paper towel drying with warm air drying. AB - OBJECTIVE: To evaluate warm air and paper towel drying for removing bacteria from washed hands. METHODS: After hands were washed with non-antibacterial soap, they were dried using warm air with and without ultraviolet light, while being rubbed or held stationary, or paper towels. Each method was performed as a randomized trial using 30 hands. RESULTS: Log colony-forming units (CFU) on palms and fingers increased significantly when hands were dried with warm air while being rubbed for 15 seconds (P < .001), and many bacteria remained at 30 seconds without ultraviolet light (P < .001) Holding hands stationary while drying significantly decreased log CFU on palms, fingers, and fingertips (P < .01 or < .001). Few CFU were detected on palms and fingers dried with ultraviolet light. Although log CFU of palms and fingers did not decrease after drying with three sheets of paper towel, those of fingertips decreased significantly (P < .001). For palms and fingers, log reductions were greater with warm air drying while holding hands stationary, paper towels, and warm air drying while rubbing hands. For fingertips, the log reduction was often greater with paper towels than with warm air. CONCLUSIONS: Holding hands stationary and not rubbing them was desirable for removing bacteria. Ultraviolet light reinforced the removal of bacteria during warm air drying. Paper towels were useful for removing bacteria from fingertips but not palms and fingers. PMID- 15796288 TI - Surveillance and infection control in an intensive care unit. AB - OBJECTIVE: To evaluate the effect of an infection control program on the incidence of hospital-acquired infection (HAI) and associated mortality. DESIGN: Prospective study. SETTING: A 2000-bed, university-affiliated hospital in Italy. PATIENTS: All patients admitted to the general intensive care unit (ICU) for more than 48 hours between January 2000 and December 2001. METHODS: The infection control team (ICT) collected data on the following from all patients: demographics, origin, diagnosis, severity score, underlying diseases, invasive procedures, HAI, isolated microorganisms, and antibiotic susceptibility. INTERVENTIONS: Regular ICT surveillance meetings were held with ICU personnel. Criteria for invasive procedures, particularly central venous catheters (CVCs), were modified. ICU care was restricted to a team of specialist physicians and nurses and ICU antimicrobial therapy policies were modified. RESULTS: Five hundred thirty-seven patients were included in the study (279 during 2000 and 258 in 2001). Between 2000 and 2001, CVC exposure (82.8% vs 71.3%; P < .05) and mechanical ventilation duration (11.2 vs 9.6 days) decreased. The HAI rate decreased from 28.7% in 2000 to 21.3% in 2001 (P < .05). The crude mortality rate decreased from 41.2% in 2000 to 32.9% in 2001 (P < .05). The most commonly isolated microorganisms were nonfermentative gram-negative organisms and staphylococci (particularly MRSA). Mortality was associated with infection (relative risk, 2.11; 95% confidence interval, 1.72-2.59; P < .05). CONCLUSION: Routine surveillance for HAI, coupled with new measures to prevent infections and a revised policy for antimicrobial therapy, was associated with a reduction in ICU HAls and mortality. PMID- 15796289 TI - Agreement for curved data. AB - An agreement problem usually involves assessing the concordance of two sets of measurements, and the problem covers a broad range of data. In practice, the observations are often curves instead of the traditional points. In this article, the agreement problem is studied for curved data. Following the rationale in constructing a correlation coefficient curve for heterocorrelaticity, an agreement curve is proposed to measure agreement as a function of the independent variable for curved data. The agreement curve overcomes the drawback when only one index is used in assessing the agreement of two measurements, and it covers all situations including the nonconstant mean, nonhomogenous variance, and the data range. A real dataset is used to demonstrate the approach and to show accurate assessment and information gained if curved data are used. PMID- 15796290 TI - Modeling anti-KLH ELISA data using two-stage and mixed effects models in support of immunotoxicological studies. AB - During preclinical drug development, the immune system is specifically evaluated after prolonged treatment with drug candidates, because the immune system may be an important target system. The response of antibodies against a T-cell-dependent antigen is recommenced by the FDA and EMEA for the evaluation of immunosuppression/enhancement. For that reason, we developed a semiquantitative enzyme-linked immunosorbent assay to measure antibodies against keyhole limpet hemocyanin. To our knowledge, the analysis of this kind of data is at this moment not yet fully explored. In this article, we describe two approaches for modeling immunotoxic data using nonlinear models. The first is a two-stage model in which we fit an individual nonlinear model for each animal in the first stage, and the second stage consists of testing possible treatment effects using the individual maximum likelihood estimates obtained in the first stage. In the second approach, the inference about treatment effects is based on a nonlinear mixed model, which accounts for heterogeneity between animals. In both approaches, we use a three parameter logistic model for the mean structure. PMID- 15796291 TI - A hierarchical Binomial-Poisson model for the analysis of a crossover design for correlated binary data when the number of trials is dose-dependent. AB - The differential reinforcement of a low-rate 72-seconds schedule (DRL-72) is a standard behavioral test procedure for screening a potential antidepressant compound. The data analyzed in the article are binary outcomes from a crossover design for such an experiment. Recently, Shkedy et al. (2004) proposed to estimate the treatments effect using either generalized linear mixed models (GLMM) or generalized estimating equations (GEE) for clustered binary data. The models proposed by Shkedy et al. (2004) assumed the number of responses at each binomial observation is fixed. This might be an unrealistic assumption for a behavioral experiment such as the DRL-72 because the number of responses (the number of trials in each binomial observation) is expected to be influenced by the administered dose level. In this article, we extend the model proposed by Shkedy et al. (2004) and propose a hierarchical Bayesian binomial-Poisson model, which assumes the number of responses to be a Poisson random variable. The results obtained from the GLMM and the binomial-Poisson models are comparable. However, the latter model allows estimating the correlation between the number of successes and number of trials. PMID- 15796292 TI - Sampling strategies for detecting rare impurities: an application in gene therapy products. AB - Detection of rare impurities in drug products presents special challenges. Replication competent adenovirus (RCA) is a rare impurity found in adenovirus based gene therapy products. Various methods are used for detection of RCAs. We primarily focus on qualitative assays. Acceptance sampling plans for detecting RCAs in batches of gene therapy products are discussed. Assuming that the number of RCA units per patient dose follows a Poisson distribution, operating characteristics (OC) of these sampling plans can be studied. The OC curves display the acceptance probabilities for batches with specific true but unknown level of RCA and can be used to assess the specificity and sensitivity of the test strategies. Application of Bayesian methodologies in the assessment of RCA levels in drug batches is also discussed. Using observed data and prior belief, a 95% credible region for the number of RCA units per patient dose can be constructed. Both classical and Bayesian calculations display the impact of sample size, sampling fraction, and assay quality on the detection of RCA. For better sensitivity, the largest possible sampling fraction that does not interfere with the logistics and the performance of the assay should be used. The choice of sample size will depend on the upper limit of the biologically safe level of RCA, the testing strategy, the desired level of sensitivity and specificity, and also on practical issues. PMID- 15796293 TI - An application of the Mantel-Haenszel statistic in process validation. AB - Discrepancies in the data of a bioequivalence study led to questions about the procedures used in the analytical laboratory. In this article we describe some statistical and graphical procedures that were used to explore the causes of a process failure for an unvalidated process. PMID- 15796294 TI - A multivariate test for similarity of two dissolution profiles. AB - A multivariate test of size a for assessing the similarity of two dissolution profiles is proposed. The inferential procedure is developed by using the approach for the common mean problem in a multivariate setup due to Halperin (1961). The performance of the proposed method is compared with Intersection Union Test as well as f2 criterion recommended by the FDA through a simulation study. All the methods are illustrated with real examples. PMID- 15796295 TI - Serially balanced designs for two sets of treatments. AB - Serially balanced designs are useful in applications in which repeated measurements of multiple samples (treatments) are being taken according to an ordered sequence. The design permits the estimation of direct treatment and carryover effects. When the samples comprise two groups, where the sequence requires samples from one group to be separated by samples from another group (e.g., when a wash step has to be performed between treated samples), then the sequence has to be arranged in such a way as to accommodate this requirement. The solution to the construction of such a serially balanced sequence is given according to the construction method given by Altan et al. (2004) for the first group of treatments in combination with the use of an F-square to dictate placement of the second group of treatments. PMID- 15796296 TI - Two-sided tolerance intervals for balanced and unbalanced random effects models. AB - A procedure for constructing two-sided beta-content, gamma-confidence tolerance intervals is proposed for general random effects models, in both balanced and unbalanced data scenarios. The proposed intervals are based on the concept of effective sample size and modified large sample methods for constructing confidence bounds on functions of variance components. The performance of the proposed intervals is evaluated via simulation techniques. The results indicate that the proposed intervals generally maintain the nominal confidence and content levels. Application of the proposed procedure is illustrated with a one-fold nested design used to evaluate the performance of a quantitative bioanalytical method. PMID- 15796297 TI - Mean particle diameters. From statistical definition to physical understanding. AB - Mean particle diameters may be used to describe and to model physical, chemical, or physiological properties of products or materials containing dispersed phases. There are different notation systems for these mean diameters, which may cause much confusion. This equally applies to their nomenclature. This article introduces the Moment-Ratio definition system and evaluates briefly the ISO definition system. The ISO system appears to have serious drawbacks. Mean particle diameters can be estimated from histograms of size distributions by Summation (M-R system) and by Integration (ISO system) over the histogram intervals. Summation tends to be more accurate than Integration and is less sensitive to low values of the lower limit of size distributions. The Summation method equations are straightforward and generally applicable. The mathematical formulas of the Integration method are difficult to apply in daily practice, and their complexity may easily hide the physical background of a mean particle diameter. A coherent nomenclature system for denoting mean particle diameters is recommended. This nomenclature system does not contain any ambiguities and clearly conveys the physical meanings of mean particle diameters. This article deals also with an empirical method to select the proper type of mean diameter to describe a physical, chemical, or physiological property of a product or material containing dispersed phases. After calculation of the mean diameters from experimental data, the relationships between the product property and these mean diameters are investigated statistically. The selection method has been illustrated by two examples. The dataset of each example consists of a set of particle size distributions and the corresponding physical product properties that are influenced by the particle sizes. Hypotheses are formulated to explain the types of selected mean diameters. Sharing results from all over the world of applications of the developed selection method will lead to a buildup of knowledge of physical meanings and application areas of the types of mean particle diameters, which will support decision making in product development. PMID- 15796298 TI - Class prediction in toxicogenomics. AB - The intent of this article is to discuss some of the complexities of toxicogenomics data and the statistical design and analysis issues that arise in the course of conducting a toxicogenomics study. We also describe a procedure for classifying compounds into various hepatotoxicity classes based on gene expression data. The methodology involves first classifying a compound as toxic or nontoxic and subsequently classifying the toxic compounds into the hepatotoxicity classes, based on votes by binary classifiers. The binary classifiers are constructed by using genes selected to best elicit differences between the two classes. We show that the gene selection strategy improves the misclassification error rates and also delivers gene pathways that exhibit biological relevance. PMID- 15796299 TI - Statistical inference for relative potency in bivariate dose-response assays with correlated responses. AB - The analysis of dose-response assays measuring two correlated responses is considered. Attention is given to statistical inference for the potency ratio. Results from a simulation study suggest that a post hoc adjustment for the correlation in parameter estimates obtained from univariate fits provides nearly as much power to detect differences in potency as a bivariate response model fit. PMID- 15796300 TI - Potential use of the scan statistic for quality control in blood product manufacturing. AB - There are minimal standards for the processing of whole blood components, and to apply those standards requires a system of quality assurances. Excessive indications of failures in compliance trigger inspections and other remedial actions, but the demarcation of what is excessive is a critical issue. Issues of low volume in some production facilities, low expected frequency of nonconformance, multiple nonindependent statistical tests, and controlling both the false-positive and false-negative rates all complicate quality assurance procedures. The scan statistic is a statistic that computes the number of events in a moving window throughout the period of risk. Monitoring plans based on scan statistics are developed to estimate the probability that a process that is under control. PMID- 15796301 TI - Double bootstrapping a tolerance limit. AB - We consider the problem of constructing tolerance limits in the context of a one way random effects model The usual parametric method for calculating tolerance intervals is based on the normality assumption. However, in practice, we frequently observe nonnormally distributed data. We propose the use of the double bootstrap (or nested bootstrap) method to estimate tolerance limits, which allows us to relax the normality assumption. PMID- 15796302 TI - Membranes for biohybrid liver support: the behaviour of C3A hepatoblastoma cells is dependent on the composition of acrylonitrile copolymers. AB - Co-polymers based on acrylonitrile, N-vinylpyrrolidone, aminoethylmethacrylate and sodium methallylsulfonate were used to prepare flat membranes by phase inversion. The surface properties of membranes were characterised by water contact angle measurements, atomic force microscopy and X-ray photoelectron spectroscopy (XPS). Membrane permeability was estimated by porosity measurements with water as test liquid. Human C3A hepatoblastoma cells were plated on these materials. Cell-material interaction was characterised by overall cell morphology, formation of focal adhesion contacts and intercellular junctions. Furthermore, cell proliferation was measured and compared with the functional activity of cells as indicated by 7-ethoxycoumarin-O-deethylation. More hydrophilic materials reduced spreading of cells, formation of focal adhesion and subsequent proliferation while homotypic cell adhesion was facilitated in correlation with stronger expressions of intercellular junctions and improved functional activity. In contrast, membranes with stronger adhesivity enhanced cell proliferation but reduced the functional activity of cells. It was concluded that the co-polymerisation of acrylonitrile with hydrophilic co-monomers, such as N-vinylpyrrolidone, could be used to tailor membrane materials for the application in biohybrid liver support systems. PMID- 15796304 TI - Effect of experimental parameters on the formation of alginate-chitosan nanoparticles and evaluation of their potential application as DNA carrier. AB - This study introduces a new procedure to prepare alginate-chitosan nanoparticles and examines several experimental parameters in relation to their formation and characteristics. Using DLS and TEM analysis, nanoparticle formation was shown to be predominantly affected by the ratio of alginate to chitosan, the molecular weight of the biopolymers and the solution pH. We report a method that results in spherical particles with mean diameters ranging from 323 nm to 1.6 microm, depending on the preparation conditions. The smallest particles were formed using lower molecular weight polymers with pH between 5.0 and 5.6 and having an alginate/chitosan weight ratio of 1:1.5. We have shown that DNA can be loaded with 60% association efficiency. Our system demonstrates suitable size, loading and release characteristics for application in drug- and gene-delivery systems. PMID- 15796303 TI - On the tissue compatibility of poly(ether imide) membranes: an in vitro study on their interaction with human dermal fibroblasts and keratinocytes. AB - Recently we have developed a novel type of membrane based on poly(ether imide) (PEI) which is considered for biomedical application. To improve its physical and biological performance it was modified by blending with poly(benzimidazole) (PBI). In the present study both membranes were characterized in terms of their physicochemical properties and in vitro tissue compatibility using human dermal fibroblasts and keratinocytes. The modified membrane (PEI*) was more hydrophilic, less porous and had an increased surface (zeta) potential. We further found that blending with PBI tends to promote cell contact, at least initially, as indicated by the improved overall cell morphology, adhesion and spreading of fibroblasts, and the development of focal adhesion complexes. The effects of fibronectin (FN) and serum coating were also beneficial when compared to pure PEI and tissue culture polystyrene (TCP), which correlates to a higher adsorption of both FN and vitronectin detected by ELISA. However, a clear tendency for homotypic cellular interaction particularly of keratinocytes was obtained in contact with membranes, which was much stronger pronounced on PEI*. Although the initial adhesion was greater on PEI*, a surprising decrease in cell growth was observed at later stages of incubation, which may be explained with the membrane-promoted cellular aggregation leading to an easier detachment from the substratum. Thus, membranes based on blends of PEI with PBI could provide a tissue compatible scaffold with lowered adhesive properties, which might be a useful tool for the transfer of cells, for example, to in vitro engineered tissue constructs. PMID- 15796305 TI - Effect of culture substrate and fibroblast growth factor addition on the proliferation and differentiation of human adipo-stromal cells. AB - The objective of this study is to investigate the proliferation and differentiation of stromal cells derived from human adipose tissues cultured on substrates with different surface properties. In addition, a similar investigation was performed on cells proliferated in different concentrations of basic fibroblast growth factor (FGF-2). The culture substrates include several polymer films with different water wettabilities, glass or a cell-culture plate, and that coated with collagen type I or IV, gelatin and FGF-2. The proliferation profiles of cells were influenced by the type of culture substrate and the growth factor concentration. A larger number of proliferated cells was observed for substrates with a water contact angle around 80 degrees, while the cell number was significantly larger for every protein-coated substrate. The rate of cell proliferation became maximal at a FGF-2 concentration of 1000 ng/ml. The FGF-2 concentration used for cell proliferation affected the differentiation profile of cells proliferated. Stromal cells, proliferated in 1 ng/ml FGF-2, were osteogenically differentiated to the strongest and fastest extent among those in other growth factor doses. The alkaline phosphatase (ALP) activity of cells increased with the increased cell number, although the activity per cell was identical, irrespective of the substrate type. The strongest adipogenic differentiation was observed for cells proliferated in 1000 ng/ml FGF-2 and the differentiation induction was maintained for a long time period. No clear dependence of the cell number on adipogenesis was observed. These findings indicate that the proliferation and differentiation of human adipose tissue derived stromal cells are influenced by the culture substrate and the concentration of FGF-2 used for proliferation. PMID- 15796306 TI - Electron beam irradiation as protection against the environmental release of recombinant molecules for biomaterials applications. AB - In biomaterials applications there exists a need to protect against the environmental release of recombinant microorganisms and transmissible genetic material and to prevent the recovery of proprietary genetic information. Irradiation technologies have long been used to eliminate microorganisms associated with spoilage and contamination and recent studies have demonstrated that moderate doses of irradiation may be used to sterilize medically important proteins without causing adverse effects in their desirable biological properties. Recombinant Escherichia coli cells expressing organophosphate hydrolase (OPH, E.C. 3.1.8.1), an important enzyme for the detection and decontamination of neurotoxic pesticides and chemical warfare agents, were subjected to electron beam irradiation to gauge its effect on enzymatic activity, cell viability and DNA recoverability. Bacterial samples were irradiated at 2, 20 and 200 kGy using a 10 MeV electron source. Irradiation levels of 2 to 20 kGy were sufficient to eliminate viable cells without affecting OPH enzymatic activity. Biologically active DNA was recovered via PCR from all samples through the 20 kGy irradiation level. While DNA was not recovered from samples at the 200 kGy exposure level, protein activity was reduced by 19 to 78%, depending on the method of cell preparation. These results demonstrate that irradiation can be effective in preventing the release of recombinant organisms intended for use in biomaterials applications without eliminating enzymatic activity and suggests that further research may indicate specific conditions whereby DNA recovery can be eliminated while retaining sufficient enzymatic activity for targeted biomaterials applications. PMID- 15796307 TI - A novel method to enhance the stability of alginate-poly-L-lysine-alginate microcapsules. AB - Implantation of microencapsulated recombinant cells is an alternative approach to gene therapy. These genetically-engineered cells enclosed in microcapsules to deliver therapeutic recombinant products have been effective in treating several murine models of human diseases. However, the most commonly used microcapsules fabricated from alginate ionically cross-linked with calcium suffer from loss of long-term mechanical stability. We now report on a method to improve their stability by introducing additional polymers to provide covalent linkages via photopolymerization. Vinyl monomers and a photoinitiator were allowed to diffuse into the initially formed calcium-alginate microcapsules. In situ photopolymerization in the presence of sodium acrylate and N-vinylpyrrolidone substantially enhanced their mechanical strength. After four months of storage in saline, > 70% of these capsules remained intact in the osmotic pressure test, while the un-modified alginate microcapsules totally disintegrated. Tests of their permeability to polyethylene glycol of different molecular weight and their ability to support cell survival showed that these properties remained unaffected by the photopolymerization. Hence, these microcapsules modified by adding a network of vinyl polymers are promising candidates to use for long-term delivery of recombinant gene products in this cell-based method of gene therapy. PMID- 15796308 TI - Quantification of initial adhesion of Enterococcus faecalis to medical grade polymers using a DNA-based fluorescence assay. AB - This paper reports on the use of a DNA-based fluorescence assay to study and quantify the initial interactions of the uropathogen Enterococcus faecalis with different polymers commonly used for the fabrication of medical devices and implants, including polyurethane (PU), silicone (SI), high-density polyethylene (HDPE), polyamide (PA), poly(methyl methacrylate) (PMMA) and polytetrafluoroethylene (PTFE). To follow the kinetics of E. faecalis adhesion, polymer samples were incubated in bacterial solution for various times and the relative concentration of adhered bacteria was obtained using two methods: commonly used CFU plate counting and a DNA quantification assay. Results obtained from DNA-based fluorescence assays showed that E. faecalis adhesion on PU is 3 times higher than that on PTFE following exposure to bacteria for 180 min. Neither surface wettability nor surface roughness of the studied polymers was found to correlate with E. faecalis adhesion, suggesting the involvement of much more complex adhesion mechanisms of bacteria onto surfaces. SEM micrographs of adhered bacteria illustrated that adhesion was different depending on the type of polymeric substrate: adhesion on PU samples was characterized by the aggregation of bacterial cells in dense clusters, as well as by the presence of fimbriae between cells and the substrate, which could explain the high adhesion to PU compared to the other polymers. This work demonstrated that the bacterial adhesion to polymers occurs at an early stage of the contact and suggests that the initial adhesion stage should be controlled, in order to prevent subsequent biofilm formation and, thus, reduce the risk of implant-associated infections. PMID- 15796309 TI - Effects of hyperbaric oxygen therapy on experimental burn wound healing in rats: a randomized controlled study. AB - A body of data supports the efficacy of hyperbaric oxygen (HBO2) therapy in the treatment of thermal burns, but the role of HBO2 in the treatment of burn injury remains a subject of controversy. The aim of this study was to evaluate possible positive effects of HBO2 on the experimental burn wound healing. Deep second degree burns were produced on the depilated backs of 70 male Wistar rats using a validated burn protocol. The animals were assigned randomly to one of two groups: 35 to the control group, which was treated with silver sulphadiazine and placebo gas, and 35 to the experimental group, which was treated with silver sulphadiazine and HBO2. The main outcome measure was wound healing, characterized by formation of post-burn edema, neoangiogenesis, number of regeneratory active follicles, necrosis staging, margination of leukocytes, and time of epithelization. A significant reduction of the post-burn edema after treatment with HBO2 (p = 0.009) was found. HBO2 had a beneficial effect on neoangiogenesis (p = 0.009). The number of preserved regeneratory active follicles was significantly higher (p = 0.009) and epithelial regeneration was more rapid in the experimental group (p = 0.048). There were no significant differences for margination of leukocytes (p = 0.55) or necrosis staging (p = 1.00). These data further support earlier conclusions that HBO2 is beneficial in the healing of burn wounds. PMID- 15796311 TI - Hyperbaric treatment of patients with carbon monoxide poisoning in the United States. AB - INTRODUCTION: Hyperbaric oxygen (HBO2) is effective therapy for carbon monoxide (CO) poisoning. In recent years, many hyperbaric physicians in the US have felt that numbers of patients referred for treatment of CO poisoning have decreased. Further, since the 2002 Weaver et al study (5), there has been discussion regarding the best treatment protocol. This study was conducted to determine numbers of patients treated with HBO2 annually over the past decade in the US and whether there is a consensus about the number of treatments per patient. MATERIALS AND METHODS: A survey was mailed to all US facilities listed in the 2001 UHMS Chamber Directory. Two subsequent mailings were sent to survey nonresponders, followed by telephone contacts. RESULTS: Of the 320 facilities listed in the directory, 10 were nonresponders, 26 had closed since publication and 80 do not treat CO poisoning, leaving 204 facilities. From 1992-2002, a total of 16,367 patients were treated with HBO2 for CO poisoning, an average of 1,488 +/- 121 patients/year (mean < or = SD). While the total number of patients treated annually did not decrease during the period studied, the number treated per facility did decline as a result of an increase in number of treating facilities. Only 46 facilities (23%) automatically give more than 1 hyperbaric treatment per CO-poisoned patient. Among those that do, 20 facilities (10%) give 3 treatments per patient. Conversely, 136 (67%) sometimes give more than one treatment and 12 facilities (8%) never retreat. CONCLUSIONS: Approximately 1,500 CO-poisoned patients are treated with HBO2 in the US annually, a number that has remained relatively constant since 1992. The majority of facilities does not routinely give more than one hyperbaric treatment, but will give repetitive treatment in certain situations. PMID- 15796310 TI - Neurological manifestations in Japanese Ama divers. AB - Repetitive breath-hold (BH) diving can lead to accumulation of nitrogen (N2) in blood and tissues, which may give rise to decompression illness (DCI). An unusual condition is "Taravana", the diving syndrome reported by Cross in the 1960s. That report generated wide discussion as to whether BH diving can cause DCI. Paulev was the first person to suggest the link between DCI and BH diving. He, a submarine medical officer developed symptoms of DCI after a series of BH dives, having proceeded the dives by spending time in a hyperbaric chamber at 20 meters for 8 minutes. Recently four professional Japanese BH divers (Ama) with histories of diving accidents were reported. Magnetic resonance imaging of these divers detected cerebral infarcts localized in the watershed areas of the brain. A survey conducted on their island revealed that many Ama divers had experienced stroke-like events. A clinical feature of DCI in BH diving is that the damage is limited to the brain. Although the mechanisms of brain damage in BH diving are unclear, N2 bubbles passing through the lungs or the heart so as to become arterialized are most likely to be the etiological factor. PMID- 15796312 TI - Scuba diving with diabetes mellitus--the UK experience 1991-2001. AB - OBJECTIVES: To survey the outcomes and practises of divers with diabetes mellitus. METHODS: Diabetic persons wishing to learn to scuba-dive or established divers who have diabetes mellitus in the UK are requested to fill in a detailed questionnaire annually. Divers are asked to provide basic epidemiological information and general diving history. Data provided by the diver's diabetic physician provided independent evidence of the diver's medical status. These data are recorded and analysed. RESULTS: Data have been gathered from 323 diabetic divers (269 male, 54 female) and 8,760 dives have been recorded over 11 years. Two fatalities were reported, both in non-insulin dependent divers. One incident of hypoglycaemia underwater in an insulin dependent diabetic diver has been reported. CONCLUSIONS: This survey showed that in the group of well-controlled diabetic divers studied, there were no serious problems due to hypoglycaemia when they dived. Long-term complications of diabetes must be excluded before a diabetic diver may be permitted to dive. PMID- 15796313 TI - Pulmonary edema in scuba divers: recurrence and fatal outcome. AB - Pulmonary edema occurring in divers using a self-contained underwater breathing apparatus (scuba) is an uncommon, probably under-reported, but potentially life threatening and recurrent condition. We report six episodes of pulmonary edema in five scuba divers seen during a period of 15 months. The four men and one woman ranged in age from 37 to 56 years and two were treated for hypertension. Symptoms were mostly dyspnea onset at depth, cough, hemoptysis and hypoxemia, which in the recurrent case led to cardiac arrest and death. All cases occurred in rather cold water. Findings on thoracic computed tomography (CT) scanning ranged from pleural effusion to ground-glass opacities restricted to a few areas of the lung. The complex underlying mechanisms that would contribute to a raised transalveolar pressure or to a disruption of the blood-gas barrier are discussed. It is important for emergency care providers to be aware of this syndrome for prompt recognition and optimal treatment. PMID- 15796314 TI - The influence of drag on human locomotion in water. AB - Propulsion in water requires a propulsive force to overcome drag. Male subjects were measured for cycle frequency, energy cost and drag (D) as a function of velocity (V), up to maximal V, for fin and front crawl swimming, kayaking and rowing. The locomotion with the largest propulsive arms and longest hulls traveled the greatest distance per cycle (d/c) and reached higher maximal V. D while locomotoring increased as a function of V, with lower levels for kayaking and rowing at lower Vs. For Vs below 1 m/s, pressure D dominated, while friction D dominated up to 3 m/s, after which wave D dominated total D. Sport training reduced the D, increased d/c, and thus lowered C and increased maximal V. Maximal powers and responses to training were similar in all types of locomotion. To minimize C or maximize V, D has to be minimized by tailoring D type (friction, pressure or wave) to the form of locomotion and velocity. PMID- 15796315 TI - A systematic review of the application of hyperbaric oxygen in the treatment of severe anemia: an evidence-based approach. AB - The treatment of severe anemia with hyperbaric oxygen (HBO2) is one of thirteen indications approved by the Hyperbaric Oxygen Therapy Committee of the Undersea and Hyperbaric Medical Society for appropriate use of the therapy (1). This paper systematically reviews the literature reporting the use of HBO2 therapy in the treatment and management of severe anemia. Increasingly, a trend to use standards of evidence-based medicine to evaluate the effectiveness of therapeutic interventions in injury and illness is productively with us in medicine today. At issue is discovery and evaluation of the best evidence available in world medical literature for evaluation of current treatment of the individual patient. The best evidence is a published randomized controlled prospective human trial; at the other end of the spectrum, the least valued evidence is a published expert opinion. In this review thirty-five publications have been reviewed as representing published results of applying HBO2 in treatment of severe anemia. Each article underwent the evidence-based evaluative grading of the American Heart Association system (AHA), the National Cancer Institute Patient Data Query system (NCI-PDQ), and the British Medical Journal's (BMJ) Clinical Evidence system. Comparative results using the three systems of evaluation are presented in tabular form for the reader. All publications report a positive result when HBO2 is delivered as treatment for severe anemia. Other alternatives other than transfusion with autologous or heterologous matched blood products are helpful but most too have not been the subject of prospective human randomized controlled trials. HBO2 may be used adjunctively with hematinics, fluorocarbons, and cell wall free polymerized hemoglobin (currently fluorocarbons and cell wall free polymerized hemoglobin are not available for routine use in the United States, but both are undergoing advanced stage clinical trials at the time of this review). PMID- 15796316 TI - Loneliness in the waiting room. PMID- 15796317 TI - The writing skills working group. PMID- 15796319 TI - On government funding for PhD studies... PMID- 15796318 TI - Unique lab broadens education options. PMID- 15796320 TI - Looking ahead to successful immunization programs. PMID- 15796321 TI - Questions and answers. Career challenges and aspirations. PMID- 15796322 TI - Evaluating best practice guidelines. AB - Clinical practice guidelines are developed with the purpose of improving patient care. The Registered Nurses Association of Ontario (RNAO) Nursing Best Practice Guidelines (BPG) Project was implemented in 1999. It has resulted in the development, implementation and evaluation of 17 BPGs with agencies in different healthcare sectors. This article describes the process, challenges and lessons learned by the team responsible for evaluating the BPGs. PMID- 15796323 TI - Planting the seeds of leadership. AB - The college of New Caledonia (CNC) in Prince George, British Columbia, believes in creating an environment where students can learn leadership skills from the beginning of their professional education. To that end, students, faculty members and administrators of CNC's health sciences and social services division worked together to engage in a campaign called "Expect a Leader." The intention of this campaign was to strengthen interdisciplinary awareness between programs and to introduce leadership qualities to future healthcare professionals. This article demonstrates how this initiative created change within six health science programs and across an entire division. The authors represent a cross disciplinary team of health professionals: former CNC students, faculty members and the administrator. This team came together to discuss leadership strategies and activities that occurred within the educational setting and in the community during the Expect a Leader campaign. The authors anticipate that the campaign will foster students' involvement in interdisciplinary leadership activities within practice settings. Readers of this article will learn about the positive effects of promoting interdisciplinary leadership campaigns across healthcare disciplines, and will gain an understanding of reasons for nurturing leadership from the beginning of the students' education. PMID- 15796324 TI - Taming the beast. The challenges for diabetes management. AB - All professionals involved in the care of individuals with diabetes should know how good blood glucose control is assessed and have at least a basic working knowledge of blood glucose management. Nursing intervention opportunities may be missed, or a well-informed diabetes patient may lose confidence in your ability to provide competent nursing care, when statements or actions reflect knowledge gaps in such basics. This article summarizes the principles of management of diabetes required for, but not limited to, the hospital setting. PMID- 15796325 TI - Research for practice, for life. PMID- 15796326 TI - The modified super-ellipsoid yield criterion for human trabecular bone. AB - Despite the importance of multiaxial failure of trabecular bone in many biomechanical applications, to date no complete multiaxial failure criterion for human trabecular bone has been developed. By using experimentally validated nonlinear high-resolution, micromechanical finite-element models as a surrogate for multiaxial loading experiments, we determined the three-dimensional normal strain yield surface and all combinations of the two-dimensional normal-shear strain yield envelope. High-resolution finite-element models of three human femoral neck trabecular bone specimens obtained through microcomputed tomography were used. In total, 889 multiaxial-loading cases were analyzed, requiring over 41,000 CPU hours on parallel supercomputers. Our results indicated that the multiaxial yield behavior of trabecular bone in strain space was homogeneous across the specimens and nearly isotropic. Analysis of stress-strain curves along each axis in the 3-D normal strain space indicated uncoupled yield behavior whereas substantial coupling was seen for normal-shear loading. A modified super ellipsoid surface with only four parameters fit the normal strain yield data very well with an arithmetic error +/-SD less than -0.04 +/- 5.1%. Furthermore, the principal strains associated with normal-shear loading showed excellent agreement with the yield surface obtained for normal strain loading (arithmetic error +/- SD < 2.5 +/- 6.5%). We conclude that the four-parameter "Modified Super Ellipsoid" yield surface presented here describes the multiaxial failure behavior of human femoral neck trabecular bone very well. PMID- 15796327 TI - Analysis of nonlinear responses of adherent epithelial cells probed by magnetic bead twisting: A finite element model based on a homogenization approach. AB - An original homogenization method was used to analyze the nonlinear elastic properties of epithelial cells probed by magnetic twisting cytometry. In this approach, the apparent rigidity of a cell with nonlinear mechanical properties is deduced from the mechanical response of the entire population of adherent cells. The proposed hyperelastic cell model successfully accounts for the variability in probe-cell geometrical features, and the influence of the cell-substrate adhesion. Spatially distributed local secant elastic moduli had amplitudes ranging from 10 to 400 Pa. The nonlinear elastic behavior of cells may contribute to the wide differences in published results regarding cell elasticity moduli. PMID- 15796328 TI - Local, three-dimensional strain measurements within largely deformed extracellular matrix constructs. AB - The ability to create extracellular matrix (ECM) constructs that are mechanically and biochemically similar to those found in vivo and to understand how their properties affect cellular responses will drive the next generation of tissue engineering strategies. To date, many mechanisms by which cells biochemically communicate with the ECM are known. However the mechanisms by which mechanical information is transmitted between cells and their ECM remain to be elucidated. "Self-assembled" collagen matrices provide an in vitro-model system to study the mechanical behavior of ECM. To begin to understand how the ECM and the cells interact mechanically, the three-dimensional (3D) mechanical properties of the ECM must be quantified at the micro-(local) level in addition to information measured at the macro-(global) level. Here we describe an incremental digital volume correlation (IDVC) algorithm to quantify large (>0.05) 3D mechanical strains in the microstructure of 3D collagen matrices in response to applied mechanical loads. Strain measurements from the IDVC algorithm rely on 3D confocal images acquired from collagen matrices under applied mechanical loads. The accuracy and the precision of the IDVC algorithm was verified by comparing both image volumes collected in succession when no deformation was applied to the ECM (zero strain) and image volumes to which simulated deformations were applied in both ID and 3D (simulated strains). Results indicate that the IDVC algorithm can accurately and precisely determine the 3D strain state inside largely deformed collagen ECMs. Finally, the usefulness of the algorithm was demonstrated by measuring the microlevel 3D strain response of a collagen ECM loaded in tension. PMID- 15796330 TI - Time-resolved DPIV analysis of vortex dynamics in a left ventricular model through bileaflet mechanical and porcine heart valve prostheses. AB - The performance of the heart after a mitral valve replacement operation greatly depends on the flow character downstream of the valve. The design and implanting orientation of valves may considerably affect the flow development. A study of the hemodynamics of two orientations, anatomical and anti-anatomical, of the St. Jude Medical (SJM) bileaflet valve are presented and compared with those of the SJM Biocor porcine valve, which served also to represent the natural valve. We document the velocity field in a flexible, transparent (LV) using time-resolved digital particle image velocimetry (TRDPIV). Vortex formation and vortex interaction are two important physical phenomena that dominate the filling and emptying of the ventricle. For the three configurations, the following effects were examined: mitral valve inlet jet asymmetry, survival of vortical structures upstream of the aortic valve, vortex-induced velocities and redirection of theflow in abidance of the Biot-Savart law, domain segmentation, resonant times of vortical structures, and regions of stagnantflow. The presence of three distinct flow patterns, for the three configurations, was identified by the location of vortical structures and level of coherence corresponding to a significant variation in the turbulence level distribution inside the LV. The adverse effect of these observations could potentially compromise the efficiency of the LV and result in flow patterns that deviate from those in the natural heart. PMID- 15796329 TI - The effect of incorporating vessel compliance in a computational model of blood flow in a total cavopulmonary connection (TCPC) with caval centerline offset. AB - BACKGROUND: The total cavopulmonary connection (TCPC), a palliative correction for congenital defects of the right heart, is based on the corrective technique developed by Fontan and Baudet. Research into the TCPC has primarily focused on reducing power loss through the connection as a means to improve patient longevity and quality of life. The goal of our study is to investigate the efficacy of including a caval offset on the hemodynamics and, ultimately, power loss of a connection. As well, we will quantify the effect of vessel wall compliance on these factors and, in addition, the distribution of hepatic blood to the lungs. METHODS: We employed a computational fluid dynamic model of blood flow in the TCPC that includes both the non-Newtonian shear thinning characteristics of blood and the nonlinear compliance of vessel tissue. RESULTS: Power loss in the rigid-walled simulations decayed exponentially as caval offset increased. The compliant-walled results, however, showed that after an initial substantial decrease in power loss for offsets up to half the caval diameter, power loss increased slightly again. We also found only minimal mixing in both simulations of all offset models. CONCLUSIONS: The increase in power loss beyond an offset of half the caval diameter was due to an increase in the kinetic contribution. Reduced caval flow mixing, on the other hand, was due to the formation of a pressure head in the offset region which acts as a barrier to flow. PMID- 15796331 TI - Three-dimensional velocity field reconstruction. AB - The problem of inter-slice magnetic resonance (MR) image reconstruction is encountered often in medical imaging applications, in such scenarios, there is a need to approximate information not captured in contiguously acquired MR images due to hardware sampling limitations. In the context of velocity field reconstruction, these data are required for visualization and computational analyses of flow fields to be effective. To provide more complete velocity information, a method has been developed for the reconstruction of flow fields based on adaptive control grid interpolation (ACGI). In this study, data for reconstruction were acquired via MRJ from in vitro models of surgically corrected pediatric cardiac vasculatures. Reconstructed velocity fields showed strong qualitative agreement with those obtained via other acquisition techniques. Quantitatively reconstruction was shown to produce data of comparable quality to accepted velocity data acquisition methods. Results indicate that ACGI-based velocity field reconstruction is capable of producing information suitable for a variety of applications demanding three-dimensional in vivo velocity data. PMID- 15796332 TI - Freezing by a flat, circular surface cryoprobe of a tissue phantom with an embedded cylindrical heat source simulating a blood vessel. AB - The effects of a thermally-significant blood vessel, simulated by an embedded acrylic tube, 4.8 mm outer diameter on the freezing field caused by a surface cryoprobe were studied experimentally in a tissue phantom. The flat, 15 mm diameter, circular cryoprobe was operated at a constant cooling rate of -8 degrees C/min by liquid nitrogen down to -60 degrees C. Water flow rates of 30 and 100 ml/min, at a constant temperature of 32.5 degrees C, were maintained in the embedded tube. The latter flow rate is typical to the lower range of blood flows in large arteries in the human body. The phase changing medium (PCM) used was a 30/70% by volume mashed potatoes flakes-water solution. Temperature measurements inside the PCM were performed in one plane perpendicular to the embedded tube, relative to which the cryoprobe was placed at 5 locations in separate experiments. This novel experimental method reduced the perturbation caused by the thermocouple junctions while facilitating rather detailed measurements of the temperature fields developing in the PCM. Results show the development of two hump-like formations on either side of the embedded tube. Freezing was retarded in the region away from the surface cryoprobe and under the tube. This accentuated the dominance of the axial effects, due to the embedded tube, over the radial ones due to the cryoprobe. Results of this study should be considered in designing protocols of cryosurgical procedures performed in the vicinity of thermally-significant blood vessels. PMID- 15796333 TI - Direct numerical simulations of micro-bubble expansion in gas embolotherapy. AB - We are currently developing a novel gas embolotherapy technique that involves the selective, acoustic vaporization of liquid perfluorocarbon droplets in or near a tumor as a possible treatment for cancer The resulting bubbles can then stick within the tumor vasculature to occlude blood flow and "starve" the tumor The potential development of high stresses during droplet vaporization is a major concern for safe implementation of this technique. No prior study, either experimentally or theoretically, addresses this important issue. In this work, the acoustic vaporization procedure of the therapy is investigated by direct numerical simulations. The nonlinear, multiphase, computational model is comprised of an ideal gas bubble surrounded by liquid inside a long tube. Convective and unsteady inertia, viscosity, and surface tension affect the bubble dynamics and are included in this model, which is solved by a novel fixed-grid, sharp-interface, moving boundary method. We assess the potential for flow-induced wall stresses to rupture the vessel or damage the endothelium during vaporization under a range of operating conditions by varying dimensionless parameters- Reynolds, Weber, and Strouhal numbers, inertial energy and initial droplet size. It is found that the wall pressure is typically highest at the start of the bubble expansion, but the maximum wall shear stress occurs at a later time. Smaller initial bubble diameters, relative to the vessel diameter, result in lower wall stresses. PMID- 15796334 TI - Effect of surrounding tissue on vessel fluid and solid mechanics. AB - There is no doubt that atherosclerosis is one of the most important health problems in the Western Societies. It is well accepted that atherosclerosis is associated with abnormal stress and strain conditions. A compelling observation is that the epicardial arteries develop atherosclerosis while the intramural arteries do not. Atherosclerotic changes involving the epicardial portion of the coronary artery stop where the artery penetrates the myocardium. The objective of the present study is to understand the fluid and solid mechanical differences between the two types of vessels. A finite element analysis was employed to investigate the effect of external tissue contraction on the characteristics of pulsatile blood flow and the vessel wall stress distribution. The sequential coupling of fluid-solid interaction (FSI) revealed that the changes of flow velocity and wall shear stress, in response to cyclical external loading, appear less important than the circumferential stress and strain reduction in the vessel wall under the proposed boundary conditions. These results have important implications since high stresses and strains can induce growth, remodeling, and atherosclerosis; and hence we speculate that a reduction of stress and strain may be atheroprotective. The importance of FSI in deformable vessels with pulsatile flow is discussed and the fluid and solid mechanics differences between epicardial and intramural vessels are highlighted. PMID- 15796335 TI - Pre-impact lower extremity posture and brake pedal force predict foot and ankle forces during an automobile collision. AB - BACKGROUND: The purpose of this study was to determine how a driver's foot and ankle forces during a frontal vehicle collision depend on initial lower extremity posture and brake pedal force. METHOD OF APPROACH: A 2D musculoskeletal model with seven segments and six right-side muscle groups was used. A simulation of a three-second braking task found 3647 sets of muscle activation levels that resulted in stable braking postures with realistic pedal force. These activation patterns were then used in impact simulations where vehicle deceleration was applied and driver movements and foot and ankle forces were simulated. Peak rearfoot ground reaction force (F(RF)), peak Achilles tendon force (FAT), peak calcaneal force (F(CF)) and peak ankle joint force (F(AJ)) were calculated. RESULTS: Peak forces during the impact simulation were 476 +/- 687 N (F(RF)), 2934 +/- 944 N (F(CF)) and 2449 +/- 918 N (F(AJ)). Many simulations resulted in force levels that could cause fractures. Multivariate quadratic regression determined that the pre-impact brake pedal force (PF), knee angle (KA) and heel distance (HD) explained 72% of the variance in peak FRF, 62% in peak F(CF) and 73% in peak F(AJ). CONCLUSIONS: Foot and ankle forces during a collision depend on initial posture and pedal force. Braking postures with increased knee flexion, while keeping the seat position fixed, are associated with higher foot and ankle forces during a collision. PMID- 15796337 TI - A multiaxial computer-controlled organ culture and biomechanical device for mouse carotid arteries. AB - Much of our understanding of vascular mechanotransduction has come from studies using either cell culture or in vivo animal models, but the recent success of organ culture systems offers an exciting alternative. In studying cell-mediated vascular adaptations to altered loading, organ culture allows one to impose well controlled mechanical loads and to perform multiaxial mechanical tests on the same vessel throughout the culture period, and thereby to observe cell-mediated vascular adaptations independent of neural and hormonal effects. Here, we present a computer-controlled perfused organ culture and biomechanical testing device designed for small caliber (50-5000 micron) blood vessels. This device can control precisely the pulsatile pressure, luminal flow, and axial load (or stretch) and perform intermittent biaxial (pressure-diameter and axial load length) and functional tests to quantify adaptations in mechanical behavior and cellular function, respectively. Device capabilities are demonstrated by culturing mouse carotid arteries for 4 days. PMID- 15796336 TI - Cartilage interstitial fluid load support in unconfined compression following enzymatic digestion. AB - Interstitial fluid pressurization plays an important role in cartilage biomechanics and is believed to be a primary mechanism of load support in synovial joints. The objective of this study was to investigate the effects of enzymatic degradation on the interstitial fluid load support mechanism of articular cartilage in unconfined compression. Thirty-seven immature bovine cartilage plugs were tested in unconfined compression before and after enzymatic digestion. The peak fluid load support decreased significantly (p < 0.0001) from 84 +/- 10% to 53 +/- 19% and from 80 +/- 10% to 46 +/- 21% after 18-hours digestion with 1.0 u/mg-wet-weight and 0.7 u/mg-wet-weight of collagenase, respectively. Treatment with 0.1 u/ml of chondroitinase ABC for 24 hours also significantly reduced the peak fluid load support from 83 +/- 12% to 48 +/- 16% (p < 0.0001). The drop in interstitial fluid load support following enzymatic treatment is believed to result from a decrease in the ratio of tensile to compressive moduli of the solid matrix. PMID- 15796338 TI - Cross-sectional profiles and volume reconstructions of soft tissues using laser beam measurements. AB - Precise geometric reconstruction is a valuable tool in the study of soft tissues biomechanics. Optical methods have been developed to determine the tissue cross section without mechanical contact with the specimen. An adaptation of the laser micrometer developed by Lee and Woo [ASME J. Biomech. Eng., 110 (2), pp. 110 114]. is proposed in which the laser-collimated beam rotates around and moves along a fixed specimen to reconstruct its cross sections and volume. Beam motion is computer controlled to accelerate data acquisition and improve beam positioning accuracy. It minimizes time-dependent shape modifications and increases global reconstruction precision. The technique is also competent for the measurement of immersed collagen matrices. PMID- 15796339 TI - A mechanical model for collagen fibril load sharing in peripheral nerve of diabetic and nondiabetic rats. AB - Peripheral neuropathy affects approximately 50% of the 15 million Americans with diabetes. It has been suggested that mechanical effects related to collagen glycation are related to the permanence of neuropathy. In the present paper, we develop a model for load transfer in a whole nerve, using a simple pressure vessel approximation, in order to assess the significant of stiffening of the collagenous nerve sheath on endoneurial fluid pressure. We also develop a fibril scale mechanics model for the nerve, to model the straightening of wavy fibrils, producing the toe region observed in nerve tissue, and also to interrogate the effects of interfibrillar crosslinks on the overall properties of the tissue. Such collagen crosslinking has been implicated in complications in diabetic tissues. Our fibril-scale model uses a two-parameter Weibull model for fibril strength, in combination with statistical parameters describing fibril modulus, angle, wave-amplitude, and volume fraction to capture both toe region and failure region behavior of whole rat sciatic nerve. The extrema of equal and local load sharing assumptions are used to map potential differences in diabetic and nondiabetic tissues. This work may ultimately be useful in differentiating between the responses of normal and heavily crosslinked tissue. PMID- 15796340 TI - Age dependency of the biaxial biomechanical behavior of human abdominal aorta. AB - BACKGROUND: The biomechanical behavior of the human abdominal aorta has been studied with great interest primarily due to its propensity to develop such maladies as atherosclerotic occlusive disease, dissections, and aneurysms. The purpose of this study was to investigate the age-related biaxial biomechanical behavior of human infrarenal aortic tissue. METHODS OF APPROACH: A total of 18 samples (13 autopsy, 5 organ donor) were harvested from patients in each of three age groups: Group 1 (<30 years old, n=5), Group 2 (between 30 and 60 years old, n=7), and Group 3 (>60 years old, n=6). Each specimen was tested biaxially using a tension-controlled protocol which spanned a large portion of the strain plane. Response functions fit to experimental data were used as a tool to guide the appropriate choice of the strain energy function W. RESULTS: Under an equibiaxial tension of 120 N/m, the average peak stretch values in the circumferential direction for Groups 1, 2, and 3 were (mean +/-SD) 1.46 +/- 0.07, 1.15 +/- 0.07, and 1.11 +/- 0.06, respectively, while the peak stretch values in the longitudinal direction were 1.41 +/- 0.03, 1.19 +/- 0.11, and 1.10 +/- 0.04, respectively. There were no significant differences between the average longitudinal and circumferential peak stretch within each group (p > 0.1), but both of these values were significantly less (p < 0.001) for Groups 2 and 3 when compared to Group 1. Patients in Group 1 were modeled using a polynomial strain energy function W, while patients in Groups 2 and 3 were modeled using an exponential form of W, suggesting an age-dependent shift in the mechanical response of this tissue. CONCLUSION: The biaxial tensile testing results reported here are, to our knowledge, the first given for the human infrarenal aorta and reinforce the importance of determining the functional form of W from experimental data. Such information may be useful for the clinician or researcher in identifying key changes in the biomechanical response of abdominal aorta in the presence of an aneurysm. PMID- 15796341 TI - Material properties and residual stress in the stage 12 chick heart during cardiac looping. AB - During the morphogenetic process of cardiac looping, the initially straight cardiac tube bends and twists into a curved tube. The biophysical mechanisms that drive looping remain unknown, but the process clearly involves mechanical forces. Hence, it is important to determine mechanical properties of the early heart, which is a muscle-wrapped tube consisting primarily of a thin outer layer of myocardium surrounding a thick extracellular matrix compartment known as cardiac jelly. In this work, we used microindentation experiments and finite element modeling, combined with an inverse computational method, to determine constitutive relations for the myocardium and cardiac jelly at the outer curvature of stage 12 chick hearts. Material coefficients for exponential strain energy density functions were found by fitting force-displacement and surface displacement data near the indenter Residual stress in the myocardium also was estimated. These results should be useful for computational models of the looping heart. PMID- 15796342 TI - A quasi-linear, viscoelastic, structural model of the plantar soft tissue with frequency-sensitive damping properties. AB - Little is known about the structural properties of plantar soft-tissue areas other than the heel; nor is it known whether the structural properties vary depending on location. Furthermore, although the quasi-linear viscoelastic (QLV) theory has been used to model many soft-tissue types, it has not been employed to model the plantar soft tissue. The structural properties of the plantar soft tissue were quantified via stress relaxation experiments at seven regions (subcalcaneal, five submetatarsal, and subhallucal) across eight cadaveric feet. The cadaveric feet were 36.9 +/- 17.4 (mean +/- S.D.) years of age, all free from vascular diseases and orthopedics disorders. All tests were performed at a constant environmental temperature of 35 degrees C. Stress relaxation experiments were performed; different loads were employed for different areas based on normative gait data. A modification of the relaxation spectrum employed within the QLV theory allowed for the inclusion of frequency-sensitive relaxation properties in addition to nonlinear elastic behavior. The tissue demonstrated frequency-dependent damping properties that made the QLV theory ill suited to model the relaxation. There was a significant difference between the elastic structural properties (A) of the subcalcaneal tissue and all other areas (p = 0.004), and a trend (p = 0.067) for the fifth submetatarsal to have less viscous damping (c1) than the subhallucal, or first, second, or third submetatarsal areas. Thus, the data demonstrate that the structural properties of the foot can vary across regions, but careful consideration must be given to the applied loads and the manner in which the loads were applied. PMID- 15796343 TI - Stabilizing PID controllers for a single-link biomechanical model with position, velocity, and force feedback. AB - In this paper we address the problem of PID stabilization of a single-link inverted pendulum-based biomechanical model with force feedback, two levels of position and velocity feedback, and with delays in all the feedback loops. The novelty of the proposed model lies in its physiological relevance, whereby both small and medium latency sensory feedbacks from muscle spindle (MS), and force feedback from Golgi tendon organ (GTO) are included in the formulation. The biomechanical model also includes active and passive viscoelastic feedback from Hill-type muscle model and a second-order low-pass function for muscle activation. The central nervous system (CNS) regulation of postural movement is represented by a proportional-integral-derivative (PID) controller. Pade approximation of delay terms is employed to arrive at an overall rational transfer function of the biomechanical model. The Hermite-Biehler theorem is then used to derive stability results, leading to the existence of stabilizing PID controllers. An algorithm for selection of stabilizing feedback gains is developed using the linear matrix inequality (LMI) approach. PMID- 15796344 TI - The effect of overshooting the target strain on estimating viscoelastic properties from stress relaxation experiments. AB - BACKGROUND: Tendon's mechanical behaviors have frequently been quantified using the quasi-linear viscoelastic (QLV) model. The QLV parameters are typically estimated by fitting the model to a single-step stress relaxation experiment. Unfortunately, overshoot of the target strain occurs to some degree in most experiments. This has never been formally investigated even though failing to measure, minimize, or compensate for overshoot may cause large errors in the estimation of parameters. Therefore, the objective of this study was to investigate the effect of overshoot on the estimation of QLV parameters. METHOD OF APPROACH: A simulated experiment was first performed to quantify the effect of different amounts of overshoot on the estimated QLV parameters. Experimental data from tendon was then used to determine if the errors associated with overshoot could be reduced when a direct fit is used (i.e., the actual strain history was used in the curve fit). RESULTS: We found that both the elastic and viscous QLV parameters were incorrectly estimated if overshoot was not properly accounted for in the fit. Furthermore, the errors associated with overshoot were partially reduced when overshoot was accounted for using a direct fit. CONCLUSIONS: A slow ramp rate is recommended to limit the amount of overshoot and a direct fit is recommended to limit the errors associated with overshoot, although other approaches such as adjusting the control system to limit overshoot could also be utilized. PMID- 15796345 TI - An algorithm for estimating acceleration magnitude and impact location using multiple nonorthogonal single-axis accelerometers. AB - Accelerations of the head are the likely cause of concussion injury, but identifying the specific etiology of concussion has been difficult due to the lack of a valid animal or computer model. Contact sports, in which concussions are a rising health care concern, offer a unique research laboratory environment. However, measuring head acceleration in the field has many challenges including the need for large population sampling because of the relatively low incidence of concussions. We report a novel approach for calculating linear acceleration that can be incorporated into a head-mounted system for on-field use during contact sports. The advantages of this approach include the use of single-axis linear accelerometers, which reduce costs, and a nonorthogonal arrangement of the accelerometers, which simplifies the design criteria for a head-mounted and helmet compatible system. The purpose of this study was to describe the algorithm and evaluate its accuracy for measuring linear acceleration magnitude and impact location using computer simulation and experimental tests with various accelerometer configurations. A 10% error in magnitude and a 10 deg error in impact location were achieved using as few as six single-axis accelerometers mounted on a hemispherical headform. PMID- 15796347 TI - Efficacy of percutaneous vertebroplasty combined with radiotherapy in osteolytic metastatic spinal tumors. AB - OBJECT: Vertebroplasty involves the percutaneous injection of polymethylmethacrylate into collapsed vertebral bodies due to hemangioma, osteoporosis, or malignant tumor. The purpose of this study was to evaluate the merits and efficacy of percutaneous vertebroplasty (PVP) combined with radiotherapy in treating patients with osteolytic metastatic spinal tumors (OMSTs). METHODS: Twenty-eight patients with OMSTs underwent PVP for the treatment of 72 vertebrae after administration of a local anesthetic or induction of general anesthesia for pain relief and spinal stabilization. Radiotherapy for suppressing tumor or inducing pain relief was performed immediately after PVP in 22 patients. Pain levels were assessed before and after the procedure by using a visual analog scale (VAS), and follow-up assessment was conducted at 1, 3, 6, and 9 months. On postoperative Day 3, marked-to-complete VAS score-based pain relief was achieved in 13 patients (48%) and moderate relief was demonstrated in 11 (41%). The mean VAS pain score was reduced from 8.2 to 3, and major complications were absent in all cases. Follow-up plain radiography was performed to assess vertebral column stability. Neither additional vertebral collapse in the treated vertebrae nor neurological deterioration was observed. CONCLUSIONS: Percutaneous vertebroplasty is a minimally invasive procedure and, when combined with radiotherapy, seems to be effective in providing pain relief and stabilization in patients with OMSTs. PMID- 15796346 TI - The changing role for neurosurgeons and the treatment of spinal deformity. AB - Spinal deformity has classically and historically been studied by those in the discipline of orthopedic surgery. This may be attributable to the orthopedic interventionalists' experience with osseous fixation for long-bone and other skeletal fractures. Neurosurgeons have maintained a long-standing interest in complex cervical spinal disorders, and their interest in the larger field of complex spinal deformity has been expanding. An understanding of spinal deformity disorders, biomechanics, bone biology, and metallurgy is necessary before clinical, teaching, and research activities can be undertaken within neurosurgery. The authors describe basic and advanced concepts of spinal deformity management with cases to illustrate teaching points. PMID- 15796348 TI - Intramedullary spinal cord tumors in patients older than 50 years of age: management and outcome analysis. AB - OBJECT: Intramedullary spinal cord tumors (IMSCTs) in the older-age adult population pose complex management issues regarding the extent of resection and functional outcome, especially in terms of quality of life. Historically, IMSCTs in the older adult population were treated with irradiation alone because it was assumed that functional recovery would be poor. The authors examined their IMSCT database and report the first large series of IMSCTs in patients older than 50 years of age. METHODS: In this retrospective clinical and chart review there were 30 cases meeting inclusion criteria drawn from databases at three different institutions. A modified McCormick Scale was used to assess functional levels in all 30 patients pre- and postoperatively. The mean age of patients in this cohort was 59.8 years (range 50-78 years), and the mean follow-up period was 10.6 years (range 2-16 years). Ependymoma was the most common tumor (83%), and 55% were located in the thoracic spine. The most common presenting symptom was sensory dysesthesia, with rare motor loss. The prodromal period to treatment was 19.4 months. Based on the McCormick Scale score at last follow-up examination 67% of patients were clinically functionally the same, 9% were worse, and 24% were improved after surgery. There were two deaths due tumor progression (both malignant tumors) and one recurrence (anaplastic astrocytoma). All three patients in whom malignant astrocytomas were diagnosed underwent postoperative radiation therapy. CONCLUSIONS: In the population of patients older than age 50 years, thoracic ependymomas are the most common IMSCTs that present characteristically with sensory symptoms. The longer prodromal period in the older adult population may reflect the fact that their diagnosis and workup is inadequate. There was no significant increase in the length of stay in the neurosurgical ward. The authors recommend motor evoked potential-guided aggressive microsurgical resection, because the long-term outcome of benign lesions is excellent (good functional recovery and no tumor recurrence). PMID- 15796350 TI - Conical working tube: a special device for endoscopic surgery of herniated lumbar discs. AB - OBJECT: The instrumentation for endoscopic discectomy continues to evolve to allow for acceptable clinical outcomes and expanding applications. The authors describe their experience in using a conical working tube equipped with a guide for angular entry of the telescope to perform endoscopic discectomy in patients with lumbar disc herniation. METHODS: Fifty-one patients (38 men and 13 women) with herniated lumbar discs underwent endoscopic lumbar surgery during the past 2.5 years at the authors' institutions. A conical working tube was inserted over sequential coaxial dilators via a muscle-splitting approach. Conventional neurosurgical instruments were used in conjunction with an angled insertion telescope. Endoscopic discectomy was performed at the L1-2 (one case), L3-4 (two cases), L4-5 (32 cases), and L5-S1 (18 cases) levels. The surgical approach was bilateral in two patients: bilateral L4-5 in one, and right L4-5 and left L5-S1 in the other. The remaining patient suffered adjacent two-level (right-sided L4-5 and L5-S1) herniations. Outcome was assessed at a mean of 11 months after surgery by using modified Macnab criteria. Outcomes were excellent in 46 (90%), fair in three (6%), and poor in two patients (4%). Complications occurred in four patients and included a dural tear in one case, postoperative neurological deterioration in two, and discitis in two; in two of these patients open surgical exploration was required. CONCLUSIONS: A separate angled entry of the telescope shortens the effective length of the working sheath and creates a better working space, thereby allowing greater instrument maneuverability and ability to use conventional neurosurgical instruments. In addition, use of this telescope in other endoscopic procedures reduces overall cost of instrumentation and treatment, and results are comparable to those reported in association with microscopic lumbar discectomy. PMID- 15796349 TI - Operative management of metastatic and malignant primary subaxial cervical tumors. AB - OBJECT: The authors describe the preoperative assessment, intraoperative strategies, and long-term outcomes in 41 consecutive patients who underwent spinal reconstruction after resection of subaxial cervical neoplasms. METHODS: Thirty-three tumors were metastatic and eight were primary. Preoperative studies included direct laryngoscopy and vertebral artery (VA) balloon occlusion tests in selected patients. Based on the tumor location, approaches included 12 anterior, 13 posterior, and 16 combined. All patients underwent aggressive intralesional resection and spinal reconstruction. In 12 patients, the VA was dissected from the periphery of the tumor, two cases of which required ligation. Fibula allograft and an anterior rigid plate fixation were most commonly used for anterior reconstruction. Posterior reconstruction was initially performed using lateral mass plates (LMPs) in 13 patients and screw/rod systems in the remaining patients. At follow up, pain level improved to mild or was absent in 39 patients (95%) who had presented with moderate or severe pain. The American Spinal Injury Association (ASIA) Scale scores were stable in 25 patients who presented with ASIA Score E and improved in 14 patients (88%) who presented with ASIA Score B, C, or D. Functional radiculopathy significantly improved in 16 (94%) of 17 patients. Complications occurred in 10 patients (24%) and included three fixation failures requiring revision. Two fixation failures involved cervical LMP screw pullout. The overall mean survival duration was 8.6 months for patients with metastatic tumors and 33.4 months for primary tumors. CONCLUSIONS: Surgery for the treatment of subaxial spine neoplasms is effective for relieving pain, encouraging functional nerve root recovery, and preserving spinal cord function with acceptable complication rates. PMID- 15796351 TI - Outcome after lumbar sequestrectomy compared with microdiscectomy: a prospective randomized study. AB - OBJECT: Microdiscectomy currently constitutes the standard treatment for herniated lumbar discs. Although limiting surgery to excision of fragments has occasionally been suggested, prospective data are lacking. Therefore, the objective of this study was to compare early outcome and recurrence rates after sequestrectomy and microdiscectomy. METHODS: Eighty-four consecutive patients 60 years of age or younger who harbored free, subligamentary, or transanular herniated lumbar discs refractory to conservative treatment were randomized to one of two treatment groups. Intraoperative parameters and findings were documented as well as pre- and postoperative symptoms such as pain, Patient Satisfaction Index (PSI), Prolo Scale score, and Short Form (SF)-36 subscale results. Follow up of at least 12 months was available in 73 patients (87%). Preoperative intergroup symptoms did not differ significantly. Surgery was significantly shorter in the sequestrectomy-treated group. Overall, low-back pain and sciatica were drastically reduced in both groups and most sensorimotor deficits improved. At 4 to 6 months, SF-36 subscales and PSI scores showed a trend in favor of sequestrectomy, leaving 3% of patients unsatisfied compared with 18% of those treated with discectomy. Outcome according to the Prolo Scale was good or excellent in 76% of discectomy-treated patients and 92% of sequestrectomy-treated patients. Reherniation occurred in four patients after discectomy (10%) and two patients after sequestrectomy (5%) within 18 months. CONCLUSIONS: Sequestrectomy does not seem to entail a higher rate of early recurrences compared with microdiscectomy. Analysis of early outcome demonstrated a trend toward superior results when sequestrectomy is performed. Although long term follow-up data are mandatory, sequestrectomy may be an advantageous alternative to standard microdiscectomy. PMID- 15796352 TI - Unilateral removal of pars interarticularis. AB - OBJECT: Lumbar radiculopathy secondary to foraminal entrapment can be treated by unilateral removal of the overlying pars interarticularis. The authors prospectively evaluated the outcome after this procedure. METHODS: Thirty-six consecutive patients underwent unilateral resection of the pars interarticularis between August 1999 and July 2002. In 18 patients acute foraminal disc herniations compressed the nerve root against the superior pedicle; in the other 18 foraminal stenosis was secondary to degenerative changes. All patients, at each visit, completed the following questionnaires: visual analog scale for overall, leg, and back pain; the Prolo Functional Economic Rating scale; and the Pain Rating Index (PRI) of the Short-Form McGill Pain Questionnaire. At 1 year, leg pain improved in 33 patients (91%). Low-back pain appeared or worsened in eight patients (22%; one in the acute herniation group and seven in the chronic degenerative group). Only one patient required lumbar fusion for pain. The Prolo economic and function scores improved in 21 (58%) and 27 (75%) patients, respectively. The PRI scores improved in 30 cases (83%). No spondylolisthesis was observed at any level at which resection had been performed. CONCLUSIONS: Unilateral removal of the pars interarticularis is effective in relieving lumbar radicular symptoms in patients with intraforaminal entrapment. The incidence of low-back pain in patients with acute foraminal disc herniations does not increase as a result of this procedure. In patients with degenerative foraminal stenosis, unilateral resection of the pars interarticularis may be a better alternative to facetectomy and segmental fusion. This procedure may be a useful tool in spine surgery. PMID- 15796353 TI - Long-term outcome in 132 consecutive patients after posterior internal fixation and fusion for Grade I and II isthmic spondylolisthesis. AB - OBJECT: The authors assessed the late outcome of patients with Meyerding Grade I and II isthmic spondylolisthesis (IS) who underwent posterior instrumentation and posterolateral fusion (PLF). Decompression and posterior internal fixation with PLF is the classic surgical treatment for painful low-grade IS. Nevertheless, outcome data are scarce and of limited value mainly because they represent small numbers of patients, short follow-up periods, or both. METHODS: The authors obtained data in the cases of 132 consecutive adult patients (mean age 40.6 years, range 15.2-69.9 years) with IS who underwent treatment between 1984 and 2003. Assessment involved analysis of responses to mailed questionnaires, clinical charts and, in cases in which unsatisfactory results were suspected, results of clinical reevaluations. Spondylolisthesis was present at L3-4 in three patients, L4-5 in 14, L3-4 in one, L3-5 in one, L5-S1 in 113, and S1-2 in one. Signs and symptoms included back and leg pain (65.3%), leg pain alone (26.3%), back pain alone (8.4%), and neurological dysfunction (18%). At a mean follow-up duration of 9.9 years (range 0.5-19.4 years), favorable results were reported for back and leg pain in 91.7 and 87.1% of patients, respectively. The mean visual analog scale scores were 2.13 for back and 1.59 for leg pain. Eighty-four patients resumed full- or part-time work, and 56.8% were capable of performing housework more easily. In 45.5% of the patients analgesic medications were not required, and 43.9% required them sporadically. The majority (63.6%) of patients reported they would undergo surgery again and recommended it to others. Thirteen (9.9%) suffered adjacent-segment morbidity, and in seven (5.3%) pseudarthrosis was documented. There were two deep and one superficial infections (2.3%). CONCLUSIONS: Posterior instrumentation and PLF, with possible neurodecompression, yielded favorable long-term results in this retrospective study of 132 patients with low-grade IS. PMID- 15796354 TI - Unilateral progressive alterations in the lumbar spine: a biomechanical study. AB - OBJECT: Lumbar radiculopathy secondary to foraminal stenosis can be treated by unilateral removal of the overlying pars interarticularis. The main concern after this procedure is spinal stability. In this study the authors evaluate the biomechanical behavior of the lumbar spine under torsional loading after unilateral progressive alterations, including resection of the pars. METHODS: Six human cadaveric L5-sacrum functional spinal units were tested while intact and then after the following sequential unilateral alterations: excision of the pars, capsulectomy, facetectomy, and discectomy. Specimens were tested in rotation by using a biomechanical testing machine, with an axial load of 280 N and torques of +/- 7.5 Nm. The specimens remained in the machine throughout testing, and the angular displacements were recorded after each set of trials. No statistically significant difference in any of the measured parameters was found between intact spines and those undergoing resection of the pars. For positive displacement (toward the side of the lesion), a significant difference from the intact condition was found after facetectomy and discectomy. For overall displacement (range of motion), spines treated with capsulectomy, facetectomy, and discectomy were significantly different from those in the intact condition. CONCLUSIONS: Unilateral removal of the pars interarticularis does not increase spinal mobility in a statistically significant fashion. The clinical implication is that the spine may not become acutely unstable after unilateral resection of the pars. PMID- 15796355 TI - A novel rat model for the study of intraosseous metastatic spine cancer. AB - OBJECT: Although metastatic spinal disease constitutes a significant percentage of all spinal column tumors, an accessible and reproducible animal model has not been reported. In this study the authors describe the technique for creating an intraosseous spinal tumor model in rats and present a functional and histological analysis. METHODS: Eighteen female Fischer 344 rats were randomized into two groups. Group 1 animals underwent a transabdominal exposure and implantation of CRL-1666 breast adenocarcinoma into the L-6 vertebral body (VB). Animals in Group 2 underwent a sham operation. Hindlimb function was tested daily by using the Basso-Beattie-Bresnahan scale. Sixteen days after tumor implantation, animals were killed and their spines were removed for histological assessment. Statistical analysis was performed using the Wilcoxon signed-rank test. By Day 15 functional analysis showed a significant decrease in motor function in Group 1 animals (median functional score 2 of 21) compared with Group 2 rats (median functional score 21 of 21) (p = 0.0217). The onset of paraparesis in Group 1 occurred within 14 to 16 days of surgery. Histopathological analysis showed tumor proliferation through the VB and into the spinal canal, with marked osteolytic activity and spinal cord compression. CONCLUSIONS: Analysis of these findings demonstrates the consistency of tumor growth in this model and validates the utility of functional testing for onset of paresis. This new rat model allows for the preclinical evaluation of novel therapeutic treatments for patients harboring metastatic spine disease. PMID- 15796356 TI - Effects of epidural hypothermic saline infusion on locomotor outcome and tissue preservation after moderate thoracic spinal cord contusion in rats. AB - OBJECT: Regionally delivered hypothermia has advantages over systemic hypothermia for clinical application following spinal cord injury (SCI). The effects of local hypothermia on tissue sparing, neuronal preservation, and locomotor outcome were studied in a moderate thoracic spinal cord contusion model. METHODS: Rats were randomized to four treatment groups and data were collected and analyzed in a blinded fashion. Chilled saline was perfused into the epidural space 30 minutes postcontusion to achieve the following epidural temperatures: 24 +/- 2.3 degrees C (16 rats), 30 +/- 2.4 degrees C (13 rats), and 35 +/- 0.9 degrees C (13 rats). Hypothermia was continued for 3 hours when a 45-minute period of rewarming was instituted. In a fourth group a moderate contusion only was induced in 14 animals. Rectal (core) and T9-10 (epidural) temperatures were measured continuously. Locomotor testing, using the Basso-Beattie-Bresnahan (Ba-Be-Br) scale, was performed for 6 weeks, and rats were videotaped for subsequent analysis. The lesion/preserved tissue ratio was calculated throughout the entire lesion cavity and the total lesion, spinal cord, and spared tissue volumes were determined. The rostral and caudal extent of gray matter loss was also measured. At 6 weeks locomotor recovery was similar in all groups (mean Ba-Be-Br Scale scores 14.88 +/- 3.71, 14.83 +/- 2.81, 14.50 +/- 2.24, and 14.07 +/- 2.39 [p = 0.77] for all four groups, respectively). No significant differences in spared tissue volumes were found when control and treatment groups were compared, but gray matter preservation was reduced in the infusion-treated groups. CONCLUSIONS: Regional cooling applied 30 minutes after a moderate contusive SCI was not beneficial in terms of tissue sparing, neuronal preservation, or locomotor outcome. This method of cooling may reduce blood flow in the injured spinal cord and exacerbate secondary injury. PMID- 15796357 TI - Spinal cord injury induction of lesional expression of profibrotic and angiogenic connective tissue growth factor confined to reactive astrocytes, invading fibroblasts and endothelial cells. AB - OBJECT: The glial scar composed of astrogliosis and extracellular matrix deposition represents a major impediment to axonal regeneration. The authors investigated the role of a novel profibrotic and angiogenic peptide connective tissue growth factor (CTGF [Hcs24/IGFBP-r2P]) in glial scar formation following spinal cord injury (SCI) in rats. METHODS: The effects of SCI on CTGF expression during glial scar maturation 1 day to 1 month post-SCI were investigated using fluorescein-activated cell sorter (FACS) immunohistochemical analysis; these findings were compared with those obtained in sham-operated (control) spinal cords. The CTGF-positive cells accumulated at the spinal cord lesion site (p < 0.0001) corresponding to areas of glial scar formation. In the perilesional rim, CTGF expression was confined to invading vimentin-positive, glial fibrillary acidic protein (GFAP)-negative fibroblastoid cells, endothelial and smooth-muscle cells of laminin-positive vessels, and GFAP-positive reactive astrocytes. The CTGF-positive astrocytes coexpressed the activation-associated intermediate filaments nestin, vimentin (> 80%), and mesenchymal scar component fibronectin (50%). CONCLUSIONS: The restricted accumulation of CTGF-reactive astrocytes and CTGF-positive fibroblastoid cells lining the laminin-positive basal neolamina suggests participation of these cells in scar formation. In addition, perilesional upregulation of endothelial and smooth-muscle CTGF expression points to a role in blood-brain barrier function modulating edema-induced secondary damage. PMID- 15796359 TI - Anatomical landmarks for the lumbar plexus on the posterior abdominal wall. AB - OBJECT: Reports describing surgical landmarks with which to identify the branches of the lumbar plexus found on the posterior abdominal wall are lacking in the English-language literature. METHODS: The authors examined 22 sides from six female and five male cadavers. Measurements were made between the branches of the lumbar plexus and various bone landmarks such as the midline vertebral bodies, supracristal plane--a horizontal line connecting the superior-most aspect of the left and right iliac crests approximating the LA-5 vertebrae--and anterior superior iliac spine (ASIS). The mean distances from the midline and as they emerged through or lateral to the psoas major muscle to the subcostal, iliohypogastric, ilioinguinal, lateral femoral cutaneous, genitofemoral, and femoral nerves, were 5.5, 6, 6.5, 6, 4.5, and 4.5 cm, respectively. At a vertical line through the midpoint between the ASIS and the midline, the subcostal, iliohypogastric, and ilioinguinal nerves were superior to the supracristal plane at mean distances of 8, 4, and 5 cm, respectively. Inferior to the supracristal plane and in a vertical line through a midpoint between the ASIS and the midline, the lateral femoral and femoral nerves were found to have mean distances of 5 and 5.5 cm, respectively. The obturator nerve had a mean distance of 3 cm lateral to the midline. Additionally, the lateral femoral cutaneous nerve had a mean distance of 1.5 cm inferomedial to the ASIS. CONCLUSIONS: A good working knowledge of the locations and anatomy of the nerves of the lumbar plexus on the posterior abdominal wall is necessary for the surgeon who operates in this region. The measurements provided herein will aid the surgeon who wishes to expose or avoid these nerves, thus precluding injury. PMID- 15796358 TI - Neuroprotection and functional recovery after application of the caspase-9 inhibitor z-LEHD-fmk in a rat model of traumatic spinal cord injury. AB - OBJECT: Apoptosis is considered one of the most significant mechanisms in the pathogenesis of neuronal damage after spinal cord injury (SCI). This form of cell death occurs via mediators known as caspases. The aim of this study was to evaluate the neuroprotective effect of the caspase-9 inhibitor, z-LEHD-fmk, in a rat model of spinal cord trauma. METHODS: Fifty-four Wistar albino rats were studied in the following three groups of 18 animals each: sham-operated controls (Group 1); trauma-only controls (Group 2); and trauma combined with z-LEHD-fmk treated animals (0.8 microM/kg; Group 3). Spinal cord injury was produced at the thoracic level by using the weight-drop technique. Responses to SCI and the efficacy of z-LEHD-fmk treatment were determined on the basis of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling staining and light and electron microscopy findings in cord tissue at 24 hours and 7 days posttrauma. Six rats from each group were also assessed for functional recovery at 3 and 7 days after SCI. This was conducted using the inclined-plane technique and a modified version of the Tarlov motor grading scale. At 24 hours postinjury, light microscopic examination of Group 2 tissue samples showed hemorrhage, edema, necrosis, polymorphonuclear leukocyte infiltration, and vascular thrombi. Those obtained in Group 3 rats at this stage showed similar features. At 24 hours postinjury, the mean apoptotic cell count in Group 2 was significantly higher than that in Group 3 (90.25 +/- 2.6 and 50.5 +/- 1.9, respectively; p < 0.05). At 7 days postinjury, the corresponding mean apoptotic cell counts were 49 +/- 2.1 and 17.7 +/- 2.6, also a significant difference (p < 0.05). Electron microscopy findings confirmed the occurrence of programmed cell death in different cell types in the spinal cord and showed that z-LEHD-fmk treatment protected neurons, glia, myelin, axons, and intracellular organelles. CONCLUSIONS: Examination of the findings in this rat model of SCI revealed that apoptosis occurs not only in neurons and astrocytes but also in oligodendrocytes and microglia. Furthermore, immediate treatment with the caspase-9 inhibitor z LEHD-fmk blocked apoptosis effectively and was associated with better functional outcome. More in-depth research of the role of programmed cell death in spinal cord trauma and further study of the ways in which caspases are involved in this process may lead to new strategies for treating SCI. PMID- 15796360 TI - Biomechanical studies of an artificial disc implant in the human cadaveric spine. AB - OBJECT: The authors compared the biomechanical performance of the human cadaveric spine implanted with a metallic ball-and-cup artificial disc at L4-5 with the spine's intact state and after anterior discectomy. METHODS: Seven human L2-S1 cadaveric spines were mounted on a biomechanical testing frame. Pure moments of 0, 1.5, 3.0, 4.5, and 6.0 Nm were applied to the spine at L-2 in six degrees of motion (flexion, extension, right and left lateral bending, and right and left axial rotation). The spines were tested in the intact state as well as after anterior L4-5 discectomy. The Maverick disc was implanted in the discectomy defect, and load testing was repeated. The artificial disc created greater rigidity for the spine than was present after discectomy, and the spine performed biomechanically in a manner comparable with the intact state. CONCLUSIONS: The results indicate that in an in vitro setting, this model of artificial disc stabilizes the spine after discectomy, restoring motion comparable with that of the intact state. PMID- 15796361 TI - Adult paravertebral pleomorphic rhabdomyosarcoma infiltrating diffusely the whole spinal axis. Case report. AB - The authors report a unique case of diffuse spinal metastatic disease due to a pleomorphic rhabdomyosarcoma (RMS) in an adult. In addition to its overall rarity, peculiar characteristics of the particular tumor included its site of origin, demonstrated radiologically as the lumbar paravertebral musculature (psoas muscle) and the transcanalicular spread into the vertebral canal, resulting in thecal compression at multiple levels. The salient clinicopathological characteristics of RMS, as they related particularly to the spine, are subsequently discussed and a short review of the major therapeutic modalities for these tumors is offered. PMID- 15796362 TI - Pituitary carcinoma with a single metastasis causing cervical spinal cord compression. Case report. AB - Pituitary carcinoma is rare, with fewer than 100 cases having been reported in the English-language literature. The diagnosis of pituitary carcinoma requires the demonstration of cerebrospinal and/or systemic metastases rather than local invasion. The lesion carries a poor prognosis; fewer than 50% of patients survive beyond 1 year after diagnosis. In this report the authors describe the case of a 68-year-old man who had undergone transsphenoidal debulking surgery and pituitary radiotherapy 4 years earlier for a pituitary adenoma. He presented with cervical cord compression due to a single metastasis from pituitary carcinoma. The authors discuss the management of this entity and review the literature for current opinion on the pathogenesis of these tumors, factors resulting in malignant transformation, and the reliability of markers that predict future malignant behavior. Evidence for the various treatment modalities is also appraised. PMID- 15796363 TI - Paragangliomas of the lumbar region. Report of two cases and review of the literature. AB - The author reports two cases of cauda equina paraganglioma (CEP) and provides a review of all previously published cases. The current radiological, neurosurgical, and pathological literature on this rare tumor is also reviewed. PMID- 15796365 TI - Intramedullary neurenteric cysts of the spine. Report of three cases. AB - Neurenteric spinal cysts are infrequently occurring dysraphic lesions that are caused by persistent or abnormal communication among neuroectoderm, notochord, and endoderm. They are generally located at the intradural, extramedullary compartment of the low cervical or upper spinal canal. They occur primarily in infants and in young adults in combination with other congenital abnormalities. The authors report on three cases of lower-thoracic and thoracolumbar intramedullary neurenteric cysts that atypically presented in adult patients in whom there was no concomitant malformation. These lesions appear notable for location, clinical presentation, intraoperative findings, and imaging features. PMID- 15796364 TI - Presacral ganglioneuromas. Report of five cases and review of the literature. AB - Ganglioneuromas are rare, benign, slow-growing tumors belonging to the neuroblastoma group. The authors describe the presentation and treatment of five consecutive patients who presented with presacral ganglioneuromas; these cases represent the largest series to date. A review of the literature is also performed. PMID- 15796366 TI - Cervical arthroplasty complicated by delayed spontaneous fusion. Case report. AB - The authors describe the case of a 55-year-old woman who presented with a left C 6 radiculopathy and neck pain and in whom there was evidence of disc/osteophyte compression of the left C-6 nerve root. The patient underwent a C5-6 anterior cervical decompression and placement of a Bryan disc prosthesis. More than 7000 cervical discs have been inserted worldwide. Postoperatively, dynamic imaging demonstrated loss of motion at the instrumented level. The patient suffered persistent neck and arm pain that was slow to resolve. Seventeen months after the initial surgery osseous fusion was observed across the interspace and posterior surface of the prosthesis. This is the first documented case of fusion occurring at the level at which cervical arthroplasty had been performed. The precise reason for this phenomenon is unclear, but potential contributing factors include patient-related issues, poor motion due to neck pain, or possibly implant-related issues. To date, this is an exceedingly rare complication and warrants careful and prolonged follow up of all arthroplasty-treated cases. PMID- 15796367 TI - Atlantooccipital transarticular screw fixation for the treatment of traumatic occipitoatlantal dislocation. Technical note. AB - The authors describe a new technique of internal atlantooccipital screw fixation involving posterior wiring and fusion for the treatment of traumatic atlantooccipital dislocation, which was performed in a 17-year-old male patient involved in a motor vehicle accident and who suffered from atlantooccipital dislocation without neurological injury. At the 6-month follow-up examination, the patient was neurologically intact with a solid occipitocervical fusion and full range of motion of the neck. PMID- 15796368 TI - Posterior atlantoaxial transpedicular screw and plate fixation. Technical note. AB - The authors describe a modified posterior atlantoaxial fixation technique for the treatment of reducible atlantoaxial instability, which can be performed simply and easily, and can decrease the risk of vessel and/or neural damage. During an 18-month period, this technique was undertaken in 11 patients with atlantoaxial instability. There was no procedure-related morbidity. The follow-up period ranged from 8 to 18 months (mean 13.2 months). Fusion was documented in all 11 patients, and there was no progression of spinal deformity. This technique can be considered an effective alternative in the treatment of atlantoaxial subluxation. PMID- 15796369 TI - Missed thoracic fractures. PMID- 15796370 TI - Cervical arthroplasty. PMID- 15796371 TI - Artificial disc. PMID- 15796372 TI - Vertebroplasty for burst fractures. PMID- 15796373 TI - Intraoperative monitoring. PMID- 15796374 TI - Gamma knife and trigeminal neuralgia. PMID- 15796375 TI - Polymorphisms in the endothelial nitric oxide synthase gene and the tendency for aneurysm rupture. PMID- 15796376 TI - Gamma knife surgery for trigeminal neuralgia: outcomes and prognostic factors. AB - OBJECT: Microvascular decompression (MVD) and percutaneous ablation surgery have historically been the treatments of choice for medically refractory trigeminal neuralgia (TN). Gamma knife surgery (GKS) has been used as an alternative, minimally invasive treatment in TN. In the present study, the authors evaluated the long-term results of GKS in the treatment of TN. METHODS: From 1996 to 2003, 151 cases of TN were treated with GKS. In this group, radiosurgery was performed once in 136 patients, twice in 14 patients, and three times in one patient. The types of TN were as follows: 122 patients with typical TN, three with atypical TN, four with multiple sclerosis-associated TN, and seven with TN and a history of a cavernous sinus tumor. In each case, the chosen radiosurgical target was located 2 to 4 mm anterior to the entry of the trigeminal nerve into the pons. The maximal radiation doses ranged from 50 to 90 Gy. The median age of the patients was 68 years (range 22-90 years), and the median time from diagnosis to GKS was 72 months (range 1-276 months). The median follow up was 19 months (range 2-96 months). Clinical outcomes and postradiosurgical magnetic resonance (MR) imaging studies were analyzed. Univariate and multivariate analyses were performed to evaluate factors that correlated with a favorable, pain-free outcome. The mean time to relief of pain was 24 days (range 1-180 days). Forty seven, 45, and 34% of patients were pain free without medication at the 1-, 2-, and 3-year follow ups, respectively. Ninety, 77, and 70% of patients experienced some improvement in pain at the 1-, 2-, and 3-year follow ups, respectively. Thirty-three (27%) of 122 patients with initial improvement subsequently experienced pain recurrence a median of 12 months (range 2-34 months) post-GKS. Among those whose symptoms recurred, 14 patients underwent additional GKS, six MVD, four glycerol injection, and one patient a percutaneous radiofrequency rhizotomy. Twelve patients (9%) suffered the onset of new facial numbness post GKS. Changes on MR images post-GKS were noted in nine patients (7%). On univariate analysis, right-sided neuralgia (p = 0.0002) and a previous neurectomy (p = 0.04) correlated with a pain-free outcome; on multivariate analysis, both rightsided neuralgia (p = 0.032) and patient age (p = 0.05) were statistically significant. New onset of facial numbness following GKS correlated with undergoing more than one GKS (p = 0.002). CONCLUSIONS: At the last follow up, GKS effected pain relief in 44% of patients. Some degree of pain improvement at 3 years post-GKS was noted in 70% of patients with TN. Although less effective than MVD, GKS remains a reasonable treatment option for those unwilling or unable to undergo more invasive surgical approaches and offers a low risk of side effects. PMID- 15796377 TI - Stereotactic gamma knife surgery for trigeminal neuralgia: detailed analysis of treatment response. AB - OBJECT: The purpose of this study was to assess the durability and completeness of pain relief in patients treated using stereotactic gamma knife surgery (GKS) for trigeminal neuralgia (TN). METHODS: Thirty-eight patients with refractory TN were treated with stereotactic GKS. All patients received a prescription radiation dose of 35, 40, or 45 Gy to the 50% isodose surface through a 4-mm collimator helmet. The group was assessed regularly based on physician-directed interviews for a median follow up of 24 months (range 6-27 months). Pain relief was classified as excellent (no pain without medication), good (well-controlled pain with continued medication), fair (decreased but residual pain with continued medication), or poor (unimproved or increased pain with the same or increased medication). Three months after treatment, pain relief was good or excellent in 71% of patients. By 24 months post-GKS, 50% of the original cohort had poor pain relief, 21% continued to have either excellent or good relief, 3% had fair relief, and 26% had not reached the 24-month follow up. Based on their status at the last follow up, 29% of patients had excellent and 16% had good pain relief. Thirty-seven percent experienced facial numbness, which was dose related. In addition, there was a significantly higher rate of complete pain relief in patients who had facial numbness following treatment (p = 0.003). CONCLUSIONS: Stereotactic GKS is an effective treatment in patients with TN; however, the durability of pain relief and the time to treatment response are limiting factors. As with other types of ablative treatment, facial numbness is strongly associated with better treatment response. PMID- 15796378 TI - Age, intracranial pressure, autoregulation, and outcome after brain trauma. AB - OBJECT: The object of this study was to investigate whether a failure of cerebrovascular autoregulation contributes to the relationship between age and outcome in patients following head injury. METHODS: Data obtained from continuous bedside monitoring of intracranial pressure (ICP), arterial blood pressure (ABP), and cerebral perfusion pressure (CPP = ABP - ICP) in 358 patients with head injuries and intermittent monitoring of transcranial Doppler blood flow velocity (FV) in the middle cerebral artery in 237 patients were analyzed retrospectively. Indices used to describe cerebral autoregulation and pressure reactivity were calculated as correlation coefficients between slow waves of systolic FV and CPP (autoregulation index [ARI]) and between ABP and ICP (pressure reactivity index [PRI]). Older patients had worse outcomes after brain trauma than younger patients (p = 0.00001), despite the fact that the older patients had higher initial Glasgow Coma Scale scores (p = 0.006). When age was considered as an independent variable, it appeared that ICP decreased with age (p = 0.005), resulting in an increasing mean CPP (p = 0.0005). Blood FV was not dependent on age (p = 0.58). Indices of autoregulation and pressure reactivity demonstrated a deterioration in cerebrovascular control with advancing age (PRI: p = 0.003; ARI: p = 0.007). CONCLUSIONS: An age-related decline in cerebrovascular autoregulation was associated with a relative deterioration in outcome in elderly patients following head trauma. PMID- 15796379 TI - The use of indomethacin in the treatment of plateau waves: effects on cerebral perfusion and oxygenation. AB - OBJECT: Plateau waves are sudden and steep increases in intracranial pressure (ICP) that can develop in patients with cerebral injuries, reduced pressure volume compensatory reserve, and preserved autoregulation. They are caused by cerebral vasodilation in response to a reduction in cerebral perfusion and are associated with increased cerebral blood volume and reduced cerebral blood flow. The authors evaluated the hypothesis that administration of indomethacin, a potent cerebral arteriolar vasoconstrictor, could interrupt the vicious cycle that occurs during plateau waves, extinguishing these waves and, ultimately, restoring cerebral perfusion and oxygenation. METHODS: Plateau waves developed in nine patients, seven with severe traumatic brain injury and two with intraparenchymal hemorrhage. One to four episodes of plateau waves per patient were treated with indomethacin (15-20 mg), which was delivered by an intravenous bolus injection. Each patient's mean arterial blood flow (MABP), ICP, cerebral perfusion pressure (CPP), and cerebral tissue PO2 were continuously monitored and the data obtained were stored in a personal computer. Each patient's jugular venous O2 saturation (SjvO2) and venoarterial difference in PCO2 were evaluated by intermittent blood sampling. During five episodes of plateau waves, middle cerebral artery flow velocities were evaluated by transcranial Doppler ultrasonography. Indomethacin extinguished all plateau waves. On average, the ICP decreased from an initial value of 58.9 +/- 11.6 mm Hg to 21.2 +/- 8.6 and 25.8 +/- 13.7 mm Hg after 5 and 10 minutes, respectively (p < 0.01). The MABP did not change significantly. As a consequence the CPP increased by 98 and 81% after 5 and 10 minutes, respectively (p < 0.01). Five and 10 minutes after indomethacin was administered, SjvO2 increased from an initial value of 50 +/- 10.5% to 62 +/- 7.6 and 59.9 +/- 9.3%, respectively (p < 0.01); the cerebral tissue PO2 increased from an initial value of 13.4 +/- 10.6 mm Hg to 23.6 +/- 9.58 and 21.9 +/- 9.2 mm Hg, respectively (p < 0.05); and the venous-arterial PCO2 decreased significantly. The mean and diastolic flow velocities increased significantly, whereas the pulsatility index decreased from 1.39 +/- 0.56 to 1.09 +/- 0.4 at 5 minutes and 1.06 +/- 0.36 at 10 minutes (p < 0.05). CONCLUSIONS: The findings confirm that plateau waves are caused by vasodilation and show that indomethacin, by constricting the cerebral arteries, is effective in extinguishing plateau waves, ultimately restoring cerebral perfusion and oxygenation. PMID- 15796380 TI - Intracerebral microdialysis in severe brain trauma: the importance of catheter location. AB - OBJECT: Intracerebral microdialysis has attracted increasing interest as a monitoring technique during neurological/neurosurgical intensive care. The purpose of this study was to compare cerebral energy metabolism, an indicator of secondary excitotoxic injury and cell membrane degradation close to focal traumatic lesions ("penumbra zones") and in remote and apparently intact brain regions of the ipsilateral and contralateral hemispheres. METHODS: The study included 22 consecutive patients with a mean age 44 +/- 17 years and an estimated postresuscitation Glasgow Coma Scale motor score less than 5. Altogether 40 microdialysis catheters with radiopaque tips were inserted. Two catheters could not be localized on postoperative computerized tomography (CT) scans and were excluded from the analysis. The perfusates were analyzed at the patient's bedside for levels of glucose, pyruvate, lactate, glutamate, and glycerol with the aid of a CMA 600 Analyzer. The positions of eight (22%) of the 36 catheters were reclassified after a review of findings on CT scans. Except for pyruvate the values of all biochemical variables and the lactate/pyruvate (L/P) ratio were significantly different in the penumbra zone when compared with mean values found in "normal" tissue ipsilateral to the parenchymal damage and in contralateral normal tissue (p < 0.001). In the penumbra zone a slow normalization of the L/P ratio and levels of glutamate and glycerol were observed. In normal tissue these parameters remained within normal limits. CONCLUSIONS: Data obtained from intracerebral microdialysis can be correctly interpreted only if the locations of the catheters as they relate to focal brain lesions are visualized. A "biochemical penumbra zone" surrounds focal traumatic brain lesions. It remains to be proven whether therapeutic interventions can protect the penumbra zone from permanent damage. PMID- 15796381 TI - Asymptomatic microbleeds in moyamoya disease: T2*-weighted gradient-echo magnetic resonance imaging study. AB - OBJECT: The aim of this study was to investigate the incidence of asymptomatic microbleeds (MBs) in patients with moyamoya disease (MMD) by using a 3-tesla magnetic resonance (MR) imaging unit. METHODS: Data on 63 patients hospitalized with MMD between 1999 and 2004 were retrospectively examined to determine the incidence of asymptomatic MBs. Gradient-echo T2*-weighted MR imaging studies obtained using 3- and 1.5-tesla units were available in 25 patients. These patients consisted of five men and 20 women, ranging in age from 17 to 66 years (mean age 41 +/- 14 years). Ischemic MMD was diagnosed in 18 patients, and hemorrhagic MMD in seven. The incidence of MBs was also evaluated using the same 3-tesla MR imaging unit in 34 healthy volunteers including seven men and 27 women, ranging in age from 18 to 71 years (mean age 33 +/- 12 years). Using the 3 tesla MR unit, asymptomatic MBs were demonstrated in 11 patients (44%); they were detected in seven patients (28%) by using the 1.5-tesla unit. In the 3-tesla MR studies in healthy individuals, MBs were found in two patients (5.8%). Based on 3 tesla MR studies, the incidence of MBs was significantly higher in patients with MMD compared with that in healthy individuals. Asymptomatic MBs were demonstrated in eight (44%) of 18 patients with ischemic MMD and three (43%) of seven patients with hemorrhagic MMD. CONCLUSIONS: Microbleeds are significantly more common in patients with MMD than in healthy individuals regardless of the disease type. The evaluation of MBs with T2*-weighted 3-tesla MR imaging might contribute to the treatment of MMD. PMID- 15796382 TI - Basilar artery dissection. AB - OBJECT: Little is understood about the clinical manifestations of basilar artery (BA) dissections, which can present with subarachnoid hemorrhage (SAH), brainstem compression, or ischemia. In any instance, the prognosis seems poorer than that for vertebral artery (VA) dissection. The authors analyzed clinical presentations and radiological features of BA dissection with and without rupture. METHODS: Between 1998 and 2003, the authors treated 10 patients (eight men and two women, ranging in age from 32-78 years; mean age 54 years) with BA dissection. Diagnosis was based on clinical and radiological findings, including those from magnetic resonance imaging and cerebral angiography studies. Of the 10 patients, five had impaired consciousness at disease onset. Among four patients presenting with SAH, two were treated conservatively and had fair outcomes without recurrent hemorrhage. The other two patients with SAH were treated using unilateral endovascular VA occlusion, but one of them subsequently suffered fatal rebleeding. A fifth patient presented with progressive signs of a mass involving the brainstem, whereas the remaining five patients showed brainstem ischemia; all were treated conservatively. Four patients could not return to their previous daily activities. CONCLUSIONS: Basilar artery dissections are rare lesions associated with significant morbidity and death. The natural course of and the treatment options for BA dissection differ considerably from those for VA dissections. Management of these lesions is controversial and difficult, and requires particular care. PMID- 15796383 TI - Upper basilar artery aneurysms: oculomotor outcomes in 163 cases. AB - OBJECT: The purpose of this study was to identify factors predictive of postoperative oculomotor nerve palsy among patients who undergo surgery for distal basilar artery (BA) aneurysms. The data can be used to estimate preoperative risk in this population. The natural history of oculomotor nerve palsy in patients with good outcomes is also defined. METHODS: The cases of 163 patients with distal BA aneurysms, who were treated surgically between 1996 and 2002, were retrospectively studied to identify factors contributing to oculomotor nerve palsy. After the data had been collected, stepwise logistic regression procedures were used to determine the predictive effects of each variable on the development of oculomotor nerve palsy and to create a scoring system. Factors that interfered with resolution of oculomotor dysfunction in patients with good outcomes were also studied. Postoperative oculomotor nerve palsy occurred in 86 patients (52.8%) with distal BA aneurysms. The following factors were associated with postoperative oculomotor dysfunction, as determined by a categorical data analysis: (1) younger patient age (p < 0.001); (2) poor admission Hunt and Hess grade (p < 0.001); (3) use of temporary arterial occlusion (p < 0.001); 4) poor Glasgow Outcome Scale score (p < 0.001); and (5) the presence of a BA apex aneurysm that projected posteriorly (p < 0.001). For patients with good outcomes, postoperative oculomotor nerve palsy resolved completely within 3 months in 31 patients (52%) and within 6 months in 47 patients (80%). The projection of the BA aneurysm was associated with incomplete oculomotor recovery at 6 months postoperatively (p = 0.019). CONCLUSIONS: The results of this study can help identify patients with a high risk for the development of oculomotor nerve palsy. This may help neurosurgeons in preoperative planning and discussions. PMID- 15796384 TI - The short form-12 and the measurement of health status in patients with cerebral aneurysms: performance, validity, and reliability. AB - OBJECT: Cerebral aneurysms can affect a patient's health status by rupture and stroke, impingement on neural structures, treatment side effects, or psychological stress. The authors assessed the performance, validity, and reliability of the Short Form-12 (SF-12), a self-administered written survey instrument, to assess health status in patients with cerebral aneurysms. METHODS: A cohort of 170 patients with cerebral aneurysms who were seen at a neurosurgery clinic underwent structured interviews including measurement of their health statuses (SF-12 physical component summary [PCS] and mental component summary [MCS]), functional status (Glasgow Outcome Scale score, modified Rankin Scale score, and Barthel Index), and mental health (Hospital Anxiety and Depression Scale score). The SF-12 scores were compared with US population norms by performing t-tests with unequal variances. The validity of the SF-12 was assessed by comparing the PCS and MCS scores with each patient's functional status and mental health scores by using rank-order methods. Inter-item reliability was assessed using the Cronbach alpha statistic. Patients with cerebral aneurysms had decreased health status PCS and MCS scores when compared with population norms (p < 0.001 for all). A history of subarachnoid hemorrhage (SAH) (p = 0.006) and previous surgical or endovascular treatment (p = 0.047) was associated with worse PCS scores. The validity of the SF-12 was supported by the relationship between the PCS and MCS scores and the patient's functional status and mental health (p < 0.001 for all). The reliability of the SF-12 was documented by the Cronbach alpha statistic (alpha = 0.76). CONCLUSIONS: Patients with cerebral aneurysms have a diminished physical and mental health status as measured using the SF-12. The presence of SAH and aneurysm treatment are associated with a worse physical health status. The SF-12 is a valid and reliable instrument for measuring health status in patients with cerebral aneurysms. PMID- 15796385 TI - Multiple aneurysms of the anterior communicating artery: radiological and surgical difficulties. AB - OBJECT: Multiple aneurysms of the anterior communicating artery (ACoA) occur rarely and have not been well investigated previously. The authors report on a consecutive series of six patients who each harbored multiple ACoA aneurysms. The radiological and surgical difficulties encountered in treating these complex and uncommon aneurysms are described and the pertinent literature is reviewed. METHODS: Between October 1996 and August 2003, the authors surgically treated 146 patients with ACoA aneurysms. Six (4.1%) of these patients harbored multiple aneurysms of the ACoA. Four of these patients were men and two were women; their ages ranged from 36 to 72 years. Five patients had two aneurysms and one patient had three. All underwent surgery performed using the pterional approach. The clinical presentations, angiograms, intraoperative difficulties, and surgical results were retrospectively analyzed. All patients had premorbid hypertension. In two cases, the aneurysms were initially misdiagnosed as a single complex aneurysm based on routine cerebral angiograms, but special angiographic views demonstrated double aneurysms. In one case, multiple ACoA aneurysms could be identified using three-dimensional (3D) computerized tomography (CT) angiography. The size of the ACoA aneurysms ranged from 3 to 12 mm (mean 5.3 mm). A total of 13 ACoA aneurysms were successfully occluded in the six patients. Four patients were discharged in good condition, and two patients died. CONCLUSIONS: Although multiple ACoA aneurysms are quite rare, the following points should be kept in mind. (1) In bilobular ACoA aneurysms, special angiographic projections and 3D CT angiography or 3D digital subtraction angiography should also be performed to obtain a correct diagnosis. The differentiation of two aneurysms from a bilobular aneurysm during the preoperative period is important for surgical planning. (2) Angiographically, detection of the ruptured aneurysm is often difficult. (3) Resection of the gyrus rectus is necessary to obtain a good operative exposure. 4) Clip selection and sequencing are important. Straight clips with short blades should be preferred to avoid narrowing of the surgeon's view and a collision between the clips. PMID- 15796386 TI - Tumors of the endolymphatic sac in patients with von Hippel-Lindau disease: implications for their natural history, diagnosis, and treatment. AB - OBJECT: Endolymphatic sac tumors (ELSTs), which often are associated with von Hippel-Lindau (VHL) disease, cause irreversible hearing loss and vestibulopathy. Clinical and imaging surveillance protocols provide new insights into the natural history, mechanisms of symptom formation, and indications for the treatment of ELSTs. To clarify the uncertainties associated with the pathophysiology and treatment of ELSTs, the authors describe a series of patients with VHL disease in whom serial examinations recorded the development of ELSTs. METHODS: Patients with VHL disease were included if serial clinical and imaging studies captured the development of ELSTs, and the patients underwent tumor resection. The patients' clinical, audiological, and imaging characteristics as well as their operative results were analyzed. Five consecutive patients (three men and two women) with a mean age at surgery of 34.8 years and a follow-up period of 6 to 18 months were included in this study. Audiovestibular symptoms were present in three patients before a tumor was evident on neuroimaging. Imaging evidence of an intralabyrinthine hemorrhage coincided with a loss of hearing in three patients. Successful resection of the ELSTs was accomplished by performing a retrolabyrinthine posterior petrosectomy (RLPP). Hearing stabilized and vestibular symptoms resolved after surgery in all patients. No patient has experienced a recurrence. CONCLUSIONS: Audiovestibular symptoms, including hearing loss, in patients with VHL disease can be the result of microscopic ELSTs. Once an ELST has been detected, it can be completely resected via an RLPP with preservation of hearing and amelioration of vestibular symptoms. Early detection and surgical treatment of small ELSTs, when hearing is still present, should reduce the incidence and severity of hearing loss, tinnitus, vertigo, and cranial nerve dysfunction, which are associated with these tumors. PMID- 15796387 TI - Decreased expression of heparanase in glioblastoma multiforme. AB - OBJECT: The authors investigated the presence of endoglycosidase heparanase in human glioblastoma multiforme (GBM) and metastatic brain tumors as well as in healthy brain tissue to explore the relationship between the biological characteristics of GBM and the role of heparanase. METHODS: Heparanase messenger (m)RNA was almost undetectable in GBMs in vivo, whereas it was frequently seen in metastatic brain tumors according to results of reverse transcription-polymerase chain reaction (RT-PCR). Immunohistochemical analysis of paraffin-embedded tissue sections showed that neoplastic cells in metastatic brain tumors, especially in cells that invaded blood vessels, exhibit intense heparanase immunoreactivity. Heparanase was present in two highly invasive glioma cell lines, U87MG and U251MG, in vitro. These cell lines did not have metastatic capability, which was tested in an experimental pulmonary metastases model in mice. The activity of heparanase in these cell lines was almost the same as that in the highly metastatic melanoma cell line B16-F1. After nude mice were inoculated with U87MG cells, however, heparanase was no longer detected in subcutaneous or intracerebral experimental glioma in vivo based on results of immunohistochemical analysis. According to results of real-time quantitative PCR, there was a 10-fold increase in heparanase mRNA in U87MG glioma cells in vitro compared with that in experimental U87MG glioma tissue in vivo in nude mice. CONCLUSIONS: These results indicate that the expression of heparanase was downregulated in GBM in vivo, which rarely metastasizes to distant organs outside the central nervous system. Heparanase is not implicated in the invasiveness of GBM to surrounding healthy brain tissue in vivo. PMID- 15796388 TI - Selective ablation of nociceptive neurons for elimination of hyperalgesia and neurogenic inflammation. AB - OBJECT: Neuropathic pain is mediated by nociceptive neurons that selectively express the vanilloid receptor 1 (VR1). Resiniferatoxin (RTX) is an excitotoxic VR1 agonist that causes destruction of VR1-positive neurons. To determine whether RTX can be used to ablate VR1-positive neurons selectively and to eliminate hyperalgesia and neurogenic inflammation without affecting tactile sensation and motor function, the authors infused it unilaterally into the trigeminal ganglia in Rhesus monkeys. METHODS: Either RTX (three animals) or vehicle (one animal) was directly infused (20 microl) into the right trigeminal ganglion in Rhesus monkeys. Animals were tested postoperatively at 1, 4, and 7 weeks thereafter for touch and pain perception in the trigeminal distribution (application of saline and capsaicin to the cornea). The number of eye blinks, eye wipes, and duration of squinting were recorded. Neurogenic inflammation was tested using capsaicin cream. Animals were killed 4 (one monkey) and 12 (three monkeys) weeks postinfusion. Histological and immunohistochemical analyses were performed. Throughout the duration of the study, response to high-intensity pain stimulation (capsaicin) was selectively and significantly reduced (p < 0.001, RTX-treated compared with vehicle-treated eye [mean +/- standard deviation]): blinks, 25.7 +/ 4.4 compared with 106.6 +/- 20.8; eye wipes, 1.4 +/- 0.8 compared with 19.3 +/- 2.5; and squinting, 1.4 +/- 0.6 seconds compared with 11.4 +/- 1.6 seconds. Normal response to sensation was maintained. Animals showed no neurological deficit or sign of toxicity. Neurogenic inflammation was blocked on the RTX treated side. Immunohistochemical analysis of the RTX-treated ganglia showed selective elimination of VR1-positive neurons. CONCLUSIONS: Nociceptive neurons can be selectively ablated by intraganglionic RTX infusion, resulting in the elimination of high-intensity pain perception and neurogenic inflammation while maintaining normal sensation and motor function. Analysis of these findings indicated that intraganglionic RTX infusion may provide a new treatment for pain syndromes such as trigeminal neuralgia as well as others. PMID- 15796389 TI - The presence of tandem endothelial nitric oxide synthase gene polymorphisms identifying brain aneurysms more prone to rupture. AB - OBJECT: It is becoming apparent that the presence of certain genetic variations (polymorphisms) may increase the individual's susceptibility to cardiovascular diseases, even in the absence of a family history. We hypothesized that brain aneurysms more prone to rupture may be identified on the basis of an individual's genotype for endothelial nitric oxide synthase (eNOS), a critical vasomodulatory protein found to be increasingly relevant to the pathobiology of aneurysms. METHODS: Patients' clinical data were recorded prospectively. Genomic DNA was isolated from blood samples obtained from individuals presenting consecutively to the Mayo Clinic with ruptured (58 patients) or unruptured (49 patients) intracranial saccular aneurysms. Using polymerase chain reaction and gene microarray technology, the following eNOS genetic polymorphisms were studied: intron-4 27-base pair variable number of tandem repeats (27 VNTR); promoter single nucleotide polymorphism (T-786C SNP); and exon-7 SNP (G894T SNP). Both groups of patients had similar demographic and clinical characteristics. For all three polymorphisms, variant alleles (p < or = 0.003) and their corresponding genotypes (p < or = 0.006) were found two to four times more frequently in patients with ruptured aneurysms than in patients with unruptured aneurysms. Strikingly, the odds ratio for presenting with a ruptured brain aneurysm among individuals demonstrating the copresence of all three variant alleles was 11.4 (95% confidence interval 1.7-75.9, p = 0.004). CONCLUSIONS: The authors have uniquely identified a set of tandem eNOS gene variations whose presence can be used to identify patients with aneurysms likely to rupture. We believe that if this finding is reproducible in a large multicenter study, in addition to known anatomical factors a rapid and cost-effective screening tool will become available to clinicians as a genetic aid to predict the risks of rupture in patients presenting with unruptured intracranial aneurysms. PMID- 15796391 TI - Distal ventriculoperitoneal shunt failure caused by silicone allergy. Case report. AB - The placement of a ventriculoperitoneal (VP) shunt is the most common form of treatment for hydrocephalus. Although allergic reactions to the silicone in shunt hardware are very rare, the authors describe a case of silicone allergy causing multiple ventricular shunt revisions. A 24-year-old man, who had undergone multiple VP shunt revisions, presented with shunt malfunction caused by allergic reaction of the tissues surrounding the shunt tubing. The patient's existing silicone-based shunt was replaced with a new polyurethane system, including the proximal and distal catheters as well as the valve mechanism. Contrary to recommendations in previous studies of silicone shunt allergies, long-term immunosuppression was not initiated. The patient was followed up for more than 8 years without recurrence of an allergic reaction to the shunt. This outcome indicates that replacing the original silicone-based shunt system with a polyurethane-based system alone is sufficient in the treatment of a silicone shunt allergy. PMID- 15796390 TI - Vascular corrosion casts mirroring early morphological changes that lead to the formation of saccular cerebral aneurysm: an experimental study in rats. AB - OBJECT: The formation of cerebral aneurysms involves complex processes and little is known about the mechanisms by which they originate, grow, and rupture. The purpose of this study was to identify early ultrastructural morphological changes that lead to the formation of experimental cerebral aneurysms. METHODS: Twenty male Sprague-Dawley rats were subjected to cerebral aneurysm induction (renal hypertension and right common carotid artery ligation); 10 intact rats served as the control group. The animals were killed after 2 months, and a vascular corrosion cast of their cerebral arteries was prepared and screened for aneurysm development by using a scanning electron microscope. Sequential morphological changes observed at the cerebral artery bifurcation in response to hemodynamic shear stress included endothelial changes, intimal pad elevation, and saccular dilation. Endothelial cell changes were the first observed morphological changes; they were followed by various degrees of artery wall dilation. No aneurysmal changes developed in any of the control rats. Of the 20 surgically treated rats, 11 displayed aneurysmal changes. In five of these animals only changes in the endothelial cell imprints could be identified. In the other six rats morphological changes in endothelial cells were associated with different stages of aneurysmal dilation. CONCLUSIONS: This is the first study to demonstrate in vivo early morphological changes that lead to the formation of cerebral aneurysms. The morphological findings indicate the principal role of endothelial cells in the pathogenesis of cerebral aneurysms and suggest that hemodynamic shear stress and blood flow patterns may precipitate these early changes. PMID- 15796392 TI - Glioblastoma multiforme metastasis to the axis. Case report. AB - Extracranial bone metastasis from glioblastoma multiforme (GBM) has rarely been reported in the literature, and most metastatic GBMs are multiple bone metastases. The authors describe the first case of a GBM with metastasis only to the axis. This 42-year-old man presented with a 2-month history of headache, nausea, vomiting, and disorientation. Magnetic resonance imaging demonstrated a right temporal tumor, which was diagnosed as a GBM based on tumor resection. The patient was treated using radiation (6000 cGy) and the intravenous administration of nimustine hydrochloride. Eighteen months thereafter, he experienced the sudden onset of neck pain. Magnetic resonance studies revealed a tumor in the axis that was diagnosed as GBM based on biopsy procedure. PMID- 15796393 TI - Brain metastasis from an adenoid cystic carcinoma of the Bartholin gland. Case report. AB - Adenoid cystic carcinoma (ACC) of the Bartholin gland is an uncommon malignant gynecological tumor. Brain metastasis from a malignant gynecological lesion is encountered rarely and the prognosis for this type of metastasis is poor. Different treatment protocols, such as resection, stereotactic radiosurgery, whole-brain radiation therapy (WBRT), and chemotherapy, are available and should be considered on an individual basis. In this article, the authors report a case of brain metastasis from an ACC of the Bartholin gland that was treated by resection and WBRT. A review of the literature did not reveal any other such case. PMID- 15796394 TI - Responsiveness of life-threatening refractory emesis to gabapentin-scopolamine therapy following posterior fossa surgery. Case report. AB - Craniotomy-associated chronic emesis can be refractory to currently approved antiemetic therapy. The authors describe a man who suffered 4 weeks of severe refractory emesis, failure to thrive, and a 40-lb weight loss after he underwent resection of a posterior fossa cholesteatoma. The patient experienced complete resolution of emesis and anorexia in response to combined gabapentin-scopolamine therapy. This case provides anecdotal evidence for the use of gabapentin scopolamine therapy in patients with chronic, refractory nausea and emesis, particularly following posterior fossa surgery, during which medullary nausea and emesis centers may be affected. PMID- 15796395 TI - Rosai-Dorfman disease with dural sinus invasion. Report of two cases. AB - Rosai-Dorfman disease (RDD) is an idiopathic proliferation of histiocytes that affects the lymph nodes. Central nervous system involvement in the absence of nodal disease is extremely rare. On neuroimaging studies, intracranial RDD appears as solitary or multiple well-circumscribed, dura-based lesions. The authors report on two cases of RDD with locally aggressive features including dural sinus invasion, which to their knowledge has never before been described. A 60-year-old woman presented with progressive dizziness and vertigo that had lasted for 1 week. Cranial computerized tomography and magnetic resonance imaging revealed an extraaxial homogeneous lobulated enhancing mass involving the right occipital lobe and the right cerebellar hemisphere. Invasion of the right transverse sinus was identified on a cerebral digital subtraction angiogram. A 59 year-old man with no prior medical illness experienced progressive weakness of both upper extremities and a partial complex seizure. Magnetic resonance imaging of his brain revealed a well-circumscribed enhancing mass in the left frontal lobe with extension to the right frontal lobe and invasion of the superior sagittal sinus. Both patients underwent resection of their brain masses. Pathological studies identified the disease as RDD in both patients. PMID- 15796396 TI - Endovascular management of traumatic and iatrogenic aneurysms of the pericallosal artery. Report of two cases. AB - Traumatic intracranial aneurysms are rare complications of closed and penetrating head injuries and may also be related to a variety of neurosurgical procedures. The primary goals in the treatment of patients harboring these lesions are early identification and intervention to prevent bleeding. Traumatic aneurysms are fragile, prone to rupture, and represent a challenging subset of vascular lesions for either surgery or endovascular therapy. Surgical approaches to aneurysms located at the pericallosal arteries are associated with higher rates of morbidity and mortality than approaches to other supratentorial aneurysms. Current endovascular treatment most often involves occlusion of the parent artery with the potential of added morbidity. The authors present their experience in the endovascular management of traumatic and iatrogenic aneurysms of the pericallosal artery achieved by primary coil embolization with parent vessel preservation. For patients harboring traumatic pericallosal aneurysms with favorable anatomical characteristics, in which the morbidity caused by parent vessel occlusion is not acceptable, endosaccular coil placement may be a valuable option. PMID- 15796397 TI - Use of an embolic protection system during endovascular recanalization of a totally occluded cervical internal carotid artery at the chronic stage. Case report. AB - A symptomatic internal carotid artery (ICA) occlusion with hemodynamic compromise was treated at its chronic stage by using an endovascular technique. An embolic protection system was used during the recanalization procedure to prevent stroke by reversing the flow from the distal ICA to the common carotid artery. The totally occluded ICA was completely recanalized through percutaneous transluminal angioplasty and stent placement. The patient's symptom (transient ischemic attack) disappeared completely after treatment with no new neurological deficit. Single-photon emission computerized tomography findings confirmed improvement of the hemodynamic compromise, and no new high-intensity spots appeared on diffusion weighted magnetic resonance imaging after treatment. This case shows that endovascular recanalization by using an embolic protection device can be considered as an alternative treatment for symptomatic ICA occlusion with hemodynamic compromise and refractoriness to antiplatelet therapy, even in the chronic stage of the illness. PMID- 15796399 TI - Use of sagittal images for localization of the subthalamic nucleus. Technical note. AB - Direct observation of the subthalamic nucleus (STN) is not always possible, and physicians at many centers rely on indirect methods that relate the position of the STN to more easily recognized structures such as the red nucleus (RN). In this paper the authors describe an indirect method of viewing the STN based on the anatomy depicted on sagittal magnetic resonance (MR) images. A review of sagittal slices appearing in standard stereotactic atlases showed that the STN lies within the angle formed by the descending internal capsule (IC) and the substantia nigra (SN). The authors' technique consists of marking the location of the STN in this nigrocapsular angle on each sagittal MR image between the RN and the lateral border of the brainstem, and transferring these points to axial MR images to build a locus of points used to describe the STN. A point is chosen in the center of this locus as the stereotactic target. Two hundred eighty-two sagittal images obtained from 71 MR imaging studies performed in 29 patients by using a fast-spin echo inversion-recovery technique were examined for the presence of the STN, the SN, and the IC. The descending IC could be detected in 97% of the slices, the SN in 95% of the slices, and the STN in 73% of the slices. This indirect method involving sagittal anatomy can be used to refine localization of the STN. PMID- 15796398 TI - A contralateral, transfrontal, extraventricular approach to stereotactic brainstem biopsy procedures. Technical note. AB - The authors report on the technique and results of stereotactic biopsy for intrinsic lateral pontine and medial cerebellar lesions via a contralateral, transfrontal, extraventricular approach. Multiplanar stereotactic magnetic resonance imaging was used to plan an intraparenchymal approach, thus limiting the number of crossed pial surfaces to one and eliminating the need to cross ependymal surfaces. After the administration of a local anesthetic agent with light intravenous sedation, six patients harboring intrinsic lateral pontine lesions underwent biopsies via this intraparenchymal approach with 100% diagnostic yield and no operative morbidity. In comparison to the ipsilateral transfrontal approach, the contralateral approach laterally expands the infratentorial area accessible during biopsy to include the lateral pons and middle cerebellar peduncle. The contralateral, transfrontal, extraventricular approach is a useful, straightforward and safe alternative to the suboccipital transcerebellar and ipsilateral, transfrontal, transtentorial routes for reaching lesions of the lateral pons and middle cerebellar peduncle. PMID- 15796400 TI - Visual loss from a dolichoectatic anterior cerebral artery. Case illustration. PMID- 15796401 TI - Intracerebral hematoma and aneurysms. PMID- 15796402 TI - Intracerebral hematoma and aneurysms. PMID- 15796403 TI - Etomidate, transfusion, and vasospasm. PMID- 15796404 TI - Etomidate, transfusion, and vasospasm. PMID- 15796405 TI - Fractions, stages, radiosurgery, and radiotherapy. PMID- 15796406 TI - Correlation of clinical and laboratory surrogate markers of immunodepletion with T cell subsets (CD4 & CD8) determined flow cytometrically in HIV infected patients: a hospital based study. AB - CD4 count is established in HIV medicine as a marker of immune depletion. However, due to financial constraints this facility is not readily available in India. Therefore, this cross sectional study in the setting of a government hospital in Pune evaluated 72 HIV infected patients for the surrogate markers of CD4 count by correlating CD counts with clinical and easily available laboratory parameters. Using a data extraction Performa, the epidemiological and clinical data of these patients was noted. Routine hematological parameters were determined. T cell subsets were studied by 2-color flow cytometry. These included CD4 counts, CD4 percentage, CD8 counts, CD8 percentage and CD4/CD8 ratio. Status of cell-mediated immunity was determined using a Multi test CMI device and Mantoux reactivity. beta2 microglobulin levels were determined in 50 of the patients. Immunological parameters were correlated with the clinical profile and other simple laboratory markers. Statistical analysis was done using regression coefficients and paired t tests. Patients who had lesser weights had lower CD4 and CD8 counts, ALC (absolute lymphocyte counts) and CD4%. Fall in hemoglobin was associated with low CD4 count, low ALC and low CD8. ALC levels correlated well with CD4 counts below 500 cells / cmm. Patients with anergic skin response to Mantoux testing and impaired cell mediated immunity had lower CD4 counts than those who had intact cell mediated immunity. beta2 microglobulin levels did not correlate well with declining immune dysfunction. However a significant inverse correlation was established between beta2 microglobulin and CD4%. PMID- 15796407 TI - In-vitro study of mouse peritoneal macrophage response to virulent and avirulent strains of mycobacteria. AB - An immunological study of pathogenesis of tuberculosis was carried out in BALB-c mice in-vitro. Peritoneal macrophages obtained from BALB-c mice were challenged with virulent (H37Rv) and avirulent (H37Ra, BCG, M. phlei) strains of mycobacteria. Activated peritoneal macrophages showed enlargement, presence of intracellular bacteria and vacuolation. These significant changes in macrophage morphology were clearly evidenced in cells infected with virulent strains of Mycobacterium tuberculosis i.e. H37Rv while being absent in cells infected with avirulent H37Ra, BCG and M. phlei. Virulent mycobacteria (H37Rv) survive the phagocytic action of macrophages by residing inside the vacuoles. The capacity of virulent and avirulent strain to stimulate TNF-alpha production from peritoneal macrophage of BALB-c mice was also examined at different time interval i.e. 1,2,4,6 and 8th day by measuring cytolytic activity of culture supernatant against murine fibroblast cell line. The pattern of highest TNF release was in case of H37Rv and least with M. phlei as measured in culture supernatant after 1,2,4,6 and 8th day. PMID- 15796408 TI - Increase in minimum inhibitory concentration to quinolones and ceftriaxone in salmonellae causing enteric fever. AB - Multidrug resistance among Salmonella typhi is well known. Reports of treatment failure in enteric fever with Ciprofloxacin made us undertake this study to determine the antibiotic susceptibility pattern of S. typhi and S. paratyphi A isolated from typhoid bacteremia cases, by disc diffusion and MIC by broth dilution method. A total of 50 strains were tested, 48 of Salmonella typhi and 2 of S. paratyphi A. The disc diffusion method was done using ampicillin, chloramphenicol, cotrimoxazole, tetracycline, ciprofloxacin, ofloxacin, cefuroxime and ceftriaxone as antibiotics. The MIC was performed using ciproloxacin, ofloxacin and ceftriaxone based on standard procedure. ACCOT resistance as determined by disc diffusion method was seen in 68% of isolates. All the strains remained susceptible to flouroquinolones cephalosporins and aminoglycosides. The MIC of ciprofloxacin, ofloxacin and ceftriaxone were in the recommended range of susceptibility as given by NCCLS, 14 (28%) strains had MIC of ciprofloxacin greater than 0.5 ug/ml with 4 strains having an MIC of 1.56 ug/ml; 25 (50%) strains had MIC of ofloxacin greater than 0.5 ug/ml and 20 (40%) strains had MIC of ceftriaxone greater than 0.5 ug/ml. The high levels of MIC of ciprofloxacin may account for treatment failure cases. The rising levels of MIC of ofloxacin and ceftriaxone in S. typhi and S. paratyphi is also of concern. We document here the emergence of high levels of MIC not only to ciprofloxacin, but also ofloxacin and ceftriaxone in S. typhi and S. paratyphi A. We recommend that MIC levels of ofloxacin and ceftriaxone should be monitored along with ciprofloxacin in treatment failure cases of enteric fever. PMID- 15796409 TI - Evaluation of quantitative buffy coat (QBC) assay and polymerase chain reaction (PCR) for diagnosis of malaria. AB - A prospective study was undertaken to compare the Polymerase Chain Reaction (PCR) and Quantitative Buffy Coat (QBC) assay with conventional Giemsa technique for diagnosis of malaria. A total of 104 samples were taken for the purpose. They comprised of fever cases suggestive of malaria (n=74) and control group, fever cases other than malaria (n=30). Peripheral blood smears were prepared by Giemsa staining and QBC assay was performed as per standard protocol. From the stored blood samples, parasite DNA was extracted and PCR was performed using P. falciparum and P. vivax specific sets of primers. The QBC assay was 100% in agreement with the Giemsa stain. Specificity of the PCR detection of P. falciparum parasites was 100%. However, sensitivity for detection of P. falciparum and P. vivax by PCR was 64.28% and 82.35% respectively. In mixed cases of malaria (n=2), PCR results were in 100% agreement with that of Giemsa. The lower sensitivity of PCR for P. falciparum could probably be due to inaccessibility of target DNA, presence of PCR inhibitors in samples and parasite strain variations. PMID- 15796410 TI - A study of Hiv/STD infections amongst commercial sex workers in Kolkata (India). Part-II: sexual behaviour, knowledge and attitude towards STD/HIV infections. AB - The sexual behaviour, knowledge, attitude and practice for prevention and control measures against STD/HIV are discussed in this paper. In all 867 sex workers, belonging to different red light areas of Kolkata viz. Sonagachi 77.28% (670), Metiabruz 14.07% (122) and 8.65% (75) from Rampurgali, Khidderpore and Bow Bazar were studied. Majority of sex workers 74.28% (644) were under thirty years of age. Around 22.26% (193) were within 20 years and 5.85%(51) were above 40 years of age. Sex workers had more than one habit but commonest were betel nut chewing in 67.59%(586) and alcohol consumption either alone or with the clients in 61.1% (529). Drug abuse (IVD) was not agreed to by any of the sex workers. Around 78.09% (677) were in the trade for last ten years, and 21.91%(190) for more than ten years. The mean duration for which sex workers remain in the trade is 6.71 years. The average number of clients visiting them were 2.67 per sex workers per day. The commonest sexual practice was normal peno-vaginal in 94.1 % (823). Peno oral sex is practiced by 32.5%(282) sex workers, of which 2.65 % (23) always, 2.31%(20) often and 27.0%(59) sometimes practiced it. Peno-rectal sex was practiced by 5.42% (47), with 2.31% (20) often and 3.11% (27) sometimes. Around 13.96% (121) sex workers participate in the group sex. The knowledge of STD/HIV infection was quite low, 49.48% (429) of CSWs had heard about STD/HIV/AIDS and around 49.6% (430) knew that STD/HIV could be prevented by condom use. PMID- 15796411 TI - Status of infectious disease markers among blood donors in a teaching hospital, Bhairahawa, western Nepal. AB - A cross-sectional group of 1548 blood donors (voluntary--554 and replacement--994 blood donors) during the period February 2001 to April 2003 at Blood Bank Centre (being run by Nepal Red Cross Society) in our Teaching Hospital in Bhairahawa, were studied and screened for antibodies to Human immunodeficiency virus type 1 and type 2 (HIV- 1 & 2), Hepatitis C virus (HCV) and hepatitis B surface antigen (HBsAg) by enzyme immunoassays and for reagin antibody for syphils by (Rapid Plasma Reagin, RPR) flocculation method. The results revealed that the overall prevalence of anti-HIV-1 was 0.13%, anti-HCV--0.13%, HBsAg positivity--0.45% and RPR reactivity for syphilis was 0.39%. HIV and HCV seropositivity was 0.20% and 0.20% respectively among replacement blood donors. None of the voluntary blood donors were found to be seropositive for HIV and HCV antibodies. HBsAg seropositivity was found to be higher among replacement (0.50%) than voluntary blood donors (0.36%). Voluntary blood donors showed a higher RPR seropositivity (0.72%) than replacement blood donors (0.20%). Males constituted 1418 (91.6%) and females were 130 (8.4%) of the total blood donors studied. The highest numbers of blood donors were in the age group of 15-29 years, both for males 902 (58.27%) and females 96 (6.2%). All the seventeen seropositive cases for all the infectious disease markers studied belonged to the sexually active and productive age group of 15-45 years and all seropositives were males except one female case (i.e HBsAg positive). One blood donor was found to be seropositive for both HIV-1 and HCV infections. None was found to be seropositive for HIV-2. Blood group status was also examined for all the blood donors tested. Maximum numbers of blood donors belonged to blood group O--516 (33.33%), followed by B 497 (32.11%), A 407 (26.29%) and AB 128 (8.27%). None of the seropositive blood donor was found to belonged to blood group AB. PMID- 15796412 TI - Annual risk of tuberculous infection in Khammam a tribal district of Andhra Pradesh. AB - A tuberculin survey to estimate annual risk of tuberculosis was conducted in Khammam tribal district during 2001-2002. A total of 8637 children were test-read -2991 without BCG scar and 5442 with BCG scar. The tests were performed using ITU PPD RT23 and the maximum size of induration was recorded at about 72 hours after the test. Based on the frequency distribution of reaction size, cut-off point for infection with tubercle bacilli was considered at 12 mm. Using this criterion, the prevalence of infection was estimated at 11.8% among children without BCG scar and 10.6% among children with BCG scar. This difference was found to be statistically insignificant. ARTI rates computed from the prevalence estimates among children without and with BCG scar were 1.6% and 1.5% respectively. It was computed as 1.5% from the prevalence in the combined group i.e. irrespective of BCG scar status. PMID- 15796413 TI - Variation in morphological characters of Culex tritaeniorhynchus (Diptera: Culicidae), a Japanese encephalitis vector at Mysore, India. AB - Studies on Culex tritaeniorhynchus Giles, were undertaken to find deviation if any, on the mean of siphonal index, comb scales, pecten teeth, denticles on apical pecten tooth, anal gills index and length of basal hair tufts on siphon in the fourth instar larvae from ground pools of Mysore city and paddy fields outside Mysore. A total of 344 larvae including 172 samples from Mysore and 172 samples from a 26 km radius outside Mysore were collected. Analysis of variance demonstrated significant differences between the two populations on 7 parameters based on morphological characteristics such as comb scales, length of siphon, width of siphon at the middle, length of anal gills, siphonal index, anal gills index and the length of basal hair tufts. So Cx. tritaeniorhynchus from different breeding places in and around Mysore city in south India can be different varieties. One variety comparatively has shorter siphon index ratio, less comb scales, less anal gills index ratio and longer basal hair tufts on siphon than the other one. Mating behavior of these varieties show that, variety from the city pools was moderate eurygamous and the one from mainly paddy fields, moderate stenogamous. PMID- 15796415 TI - Knowledge of human immunodeficiency virus and post exposure prophylaxis amongst junior doctors. PMID- 15796414 TI - Prevalence of urinary schistosomiasis in school children in Enugu, Nigeria. PMID- 15796416 TI - Antibiotic resistance pattern in aerobic bacterial isolates in Kangra (H.P.). PMID- 15796417 TI - Should elective cesarean birth be an accepted option for women? PMID- 15796419 TI - Best practices in perinatal and neonatal nursing: vacuum-assisted vaginal delivery. PMID- 15796418 TI - Patient safety: examples in the NICU. PMID- 15796420 TI - Alterations in drug handling in the pregnant woman and neonate. PMID- 15796421 TI - Improving perinatal and neonatal patient safety: The AHRQ patient safety indicators. AB - This study reviews the development and implementation of the Agency for Healthcare Research and Quality (AHRQ) patient safety indicators (PSIs). The genesis of the use of administrative data as a tool to combat safety problems is presented, and how indicators were constructed using various administrative codes. Examples of how the PSIs are being used to identify potential safety problems within the general population are presented, with a special emphasis on how these are being used within the perinatal and neonatal arena to understand current issues within that subpopulation. Results from studies within the general population and targeted at perinatal and neonatal patients are presented. Finally, suggestions are discussed for clinicians to use the AHRQ PSIs as one of their early warning tools for potential safety-related problems. PMID- 15796422 TI - Failure to rescue: implications for evaluating quality of care during labor and birth. AB - Failure to rescue is an indicator that has been used to measure quality of care for surgical patients by evaluating the number of patients who die after developing postoperative complications. There are 2 key components of failure to rescue: (a) careful surveillance and timely identification of complications and (b) taking action by quickly initiating appropriate interventions and activating a team response. This concept has not been explored as a potential method to evaluate quality of intrapartum care. In obstetrics, complications leading to death are relatively rare because mothers and infants are generally healthy. Thus, there are not large numbers of maternal or infant deaths in individual hospitals or healthcare systems that allow the types of statistical analyses that have been previously used to measure failure to rescue rates. With modifications in the measurement process for failure to rescue in this population, there are direct implications for perinatal patient safety and lessons to be learned. A new use of the failure to rescue concept in a population not previously considered is proposed. PMID- 15796423 TI - Perinatal Patient Safety Project: a multicenter approach to improve performance reliability at Kaiser Permanente. AB - High-reliability organizations, including high-reliability perinatal units, are built on a solid foundation of timely communication and collegial teamwork to maintain patient safety as a top priority. In these units, concise, real-time, and constant communication is valued. Teamwork and collegiality characterize the professional relationships and hierarchy is minimized. The Perinatal Patient Safety Project utilized human factors techniques and systems improvements to create high-reliability perinatal units within a 1-year pilot project. This article focuses on the approaches used to improve communication in 4 Kaiser Permanente medical centers participating in this project. PMID- 15796424 TI - Patient safety and teamwork in perinatal care: resources for clinicians. AB - Recent data reveal communication issues and organizational culture to be key factors in adverse perinatal outcomes. Hierarchical communication is common in healthcare and can be a significant impediment to safe care. Principles of teamwork employed by other industries, such as aviation and the military, can be appropriately applied to healthcare. This article provides a brief introduction to Crew Resource Management as well as a listing of print, multimedia, and Web resources for clinicians interested in promoting cultural change and effective teamwork. PMID- 15796425 TI - Safety: when infants and parents are research subjects. AB - Patient safety is a central concern in nursing. Unlike other areas of patient safety, safety in research is particularly important because research is not part of standard care and participation is voluntary. Issues related to safety in research are especially pertinent to high-risk infants, because of the nature of parental (or legal guardian) consent and because children are considered a vulnerable group requiring special protection from research risks. Nurses must be aware of safety in research whether independently conducting research, employed by a research project, or caring for patients who are research subjects. This article reviews safety issues and policies, processes, and ethical guidelines designed to protect infants and children who are research subjects. PMID- 15796426 TI - Enteral feeding for high-risk neonates: a digest for nurses into putative risk and benefits to ensure safe and comfortable care. AB - Enteral feeding is considered a relatively safe method of providing nutritional support to high-risk neonates. Nonetheless, there are associated risks, which can be classified as follows: factors to consider before initiating enteral feeding; feeding tube placement; delivery of milk feedings; and gastrointestinal, environmental, and technical factors. For each classification, this article highlights adverse consequences and synthesizes the literature for evidence-based nursing practice recommendations, which are summarized in the "Conclusion" section. Many gaps are identified in the research literature, and directions for future research are described to ensure safe and comfortable care for high-risk neonates receiving enteral feedings. PMID- 15796428 TI - [Don't lose your precious confidence]. PMID- 15796427 TI - Maximizing patient safety: filter needle use with glass ampules. AB - Particle contamination of medications obtained from glass ampules can pose serious hazards to patients. Particle contamination may be reduced by using a filter needle when obtaining medication from glass ampules prior to administration. The purpose of this article is 2-fold, to educate nursing staff caring for neonates about (1) the potential risk and resultant complications related to particle contamination and the benefit of utilizing a filter needle when obtaining medication from glass ampules, prior to administration of the medication to patients, and (2) the construction of a unit-specific filter needle protocol. The article promotes the continued change in nursing practice regarding nurses erring on the side of safety, with the ultimate goal of improving patient safety and outcome. PMID- 15796429 TI - [Prophylaxis and treatment of postoperative deep vein thrombosis and pulmomary thromboembolism]. PMID- 15796430 TI - [Pathogenesis, diagnosis, and treatment of postoperative deep vein thrombosis]. AB - Most postoperative deep vein thrombosis (DVT) arises from the venous systems of the pelvis and lower extremities, especially the soleal veins. Embolization of venous thrombi is related to the size and location of thrombi and movement of the lower limbs and commonly occurs within 1 week from the onset of formation. There are three steps in the final diagnosis of DVT: probable diagnosis by anamnesis and physical examination; screening diagnosis using quantitative tests; and definitive diagnosis using imaging tests. To determine embolic sources, venous echography, which is noninvasive and convenient, is the first choice. Therapeutic methods are selected based on thrombi extent and time after formation. Anticoagulant therapy is indicated in all cases except in patients with possible bleeding tendency and continues for 3 months or more. Among the endovascular therapies, catheter-directed thrombolysis is a more effective approach when combined with a temporary vena cava filter than operative thrombectomy. Although the prevention of DVT is mandatory for surgeons, it is difficult to avoid venous thromboembolism completely. Systemic early diagnosis and emergent therapeutic strategies for venous thromboembolism are clinically effective and promising. PMID- 15796431 TI - [Postoperative venous thrombosis in general surgery patients and perioperative prophylaxis]. AB - In a population of general surgery patients in Western countries, there was a 19% incidence of deep vein thrombosis (DVT) and a 1.6% incidence of pulmonary embolism (PE), with 0.9% of patients experiencing fatal PE. In Japan, there was a 15.8% incidence of DVT and a 0.34% incidence of PE, with 0.08% of patients experiencing fatal PE in a population of abdominal surgery patients. The incidences of PE and fatal PE in our department were 0.11% and 0.03%, respectively. We started to use intermittent pneumatic compression (IPC) for the prophylaxis of postoperative PE in 1999 and then added elastic stockings in 2002 and low-dose unfractionated heparin (LDUH) in 2003 for prophylaxis. The incidence of PE has dropped and that of fatal PE has become 0% with the use of such prophylactic measures. When the risk of venous thromboembolism of the 15 patients who experienced PE in our department were assessed using the Japanese Guidelines for Prevention of Venous Thromboembolism, 13 patients were assessed as high risk and 2 as low risk. The mean age and mean body mass index of the 15 patients were 54 yeas old and 24.8, respectively, and PE was not limited to obese or elderly patients. Such findings appear to indicate the difficulty of risk assessment for PE. Therefore we started to use IPC, elastic stockings, and LDUH for the prophylaxis of PE and DVT for all general surgery patients from April 2004. These prophylactic measures are recommended for the highest-risk patients in the Japanese Guidelines for Prevention of Venous Thromboembolism. There have so far been no serious bleeding complications with the administration of LDUH. We will continue to observe the effects of prophylaxis and the risk of bleeding. PMID- 15796432 TI - [Postoperative pulmonary thromboembolism and its prevention from the perspective of the establishment of Japanese guidelines]. AB - The incidence of venous thromboembolism (VTE) is rapidly increasing in Japan. Ideally, the guidelines for its prevention should be based on solid evidence, but there are few clinical data on the prevalence and prophylaxis of VTE in Japan. Comprehensive, standard guidelines for the prevention of VTE were recently established by applying the risk-prophylaxis relationship of the sixth American College of Chest Physicians (ACCP) consensus recommendations. Since patients undergoing cancer surgery make up a relatively homogenous, "highest-risk" group, we could precisely evaluate the difference in VTE risk between Japan and the West. The risk of clinical PE in Japanese cancer surgery was found to be one or two risk levels lower than that in the West. Therefore the Japanese risk stratification was set to be one level lower than that in the sixth ACCP guidelines. For prophylaxis, the recommendations of the Japanese guidelines are basically identical to those of the sixth ACCP guidelines. Since the present guidelines are not sufficiently based on Japanese evidence, they should be reviewed in a prospective, nationwide epidemiologic study. In this review article, the theoretical background of and future perspectives on the Japanese guidelines are discussed. PMID- 15796433 TI - [Therapy of acute pulmonary thromboembolism from the physician's standpoint]. AB - The therapy of acute pulmonary thromboembolism (APTE) is based on the clinical grade and ranges from ambulant therapy with anticoagulation, to thrombolysis, inferior vena cava (IVC) filtration, and catheter thrombectomy. In the absence of contraindications, initial treatment of APTE should consist of parenteral anticoagulation with unfractionated heparin. Long-term anticoagulation therapy, usually with warfarin, should be administered according to the individual risk profile of the patient. Thrombolytic therapy may be appropriate for patients with massive APTE with cardiac shock, syncope, etc. Similarly, thrombolysis has been reported to be effective in submassive APTE with right ventricular overload on echocardiography. IVC filters should be reserved for APTE with deep vein thrombosis (DVT) in which there are absolute contraindications to anticoagulation, recurrent thromboemboli despite therapeutic anticoagulation, and status after surgical thrombectomy. Relative indications for IVC filters that require individualized decision making include proximal DVT, especially with free floating thrombi or in patients with limited cardiopulmonary reserve. For patients with massive APTE with contraindications to anticoagulation or in whom anticoagulation is uneffective, transcatheter aspiration with catheterization or fragmentation using a guidewire and rotating pig-tail catheter can be used. In addition, cardiopulmonary management such as supplemental oxygen, catecholamine administration, percutaneous cardiopulmonary support, etc. may be necessary for individual patients. PMID- 15796434 TI - [Pulmonary embolectomy for acute massive pulmonary thromboembolism]. AB - Acute pulmonary thromboembolism is a frequently lethal and acute-onset in hospital complication after surgery. Absolute indications for surgical embolectomy are acute massive pulmonary embolism with deep shock, refractory circulatory collapse, and continuous hypoxemia. Although thrombolytic therapy is indicated for patients with pulmonary thromboembolism with right ventricular overload, it is contraindicated for patients after major surgery or with stroke due to the high risk of rebleeding. Therefore surgical embolectomy should be considered in those patients. Pulmonary embolectomy relieves the right ventricular overload, and immediate restoration of right ventricular function contributes to the recovery of hemodynamics. A recent study revealed improved outcome for massive pulmonary embolism with early diagnosis with multidetector row computed tomography, risk stratification using echocardiography, and surgical embolectomy. Surgical pulmonary thromboembolectomy should be considered for critically ill patients with massive pulmonary thromboembolism. PMID- 15796436 TI - [Evaluating physician's work; lessons from U.S. resource-based relative value system]. PMID- 15796435 TI - [Surgical treatment for chronic pulmonary thromboembolism]. AB - Chronic pulmonary thromboembolism is a serious disorder because hypoxemia and pulmonary hypertension progress, finally resulting in respiratory and right-side heart failures. We evaluated the results of surgical treatment in patients in whom circulatory arrest was induced under profound hypothermia. Between 1995 and April 2004, 89 cases were surgically treated. The pathologic condition of these patients was classified as being degree III or greater in the NYHA classification. Following a median sternotomy, profound hypothermia was induced using cardiopulmonary bypass, and pulmonary thromboendarterectomy in the bilateral pulmonary arteries was performed under intermittent circulatory arrest. Of the 89 patients, 4 of 5 who underwent emergent surgery died after postoperatively. Among 84 patients who underwent elective surgery, 7 died of respiratory and cardiac failure. Clinical symptoms were markedly improved by surgery in 73 patients. Because this disease is resistant to medical treatment, pulmonary thromboendarterectomy using intermittent circulatory arrest under profound hypothermia is effective in treating patients with chronic thromboembolic pulmonary hypertension. PMID- 15796437 TI - [Evaluation of physician's work: lesson from the common classification of medical activities (CCAM) in France]. PMID- 15796438 TI - [What we can learn from a case of medical malpractice]. PMID- 15796439 TI - [Palliative operation for rectal cancer-perineal resection of the rectum with loop colostomy]. PMID- 15796440 TI - Doctor, I don't want chemotherapy! AB - The primary reasons for Mrs. S's refusal of treatment related to the emotional distress of an unexpected treatment recommendation (chemotherapy) and inaccurate beliefs regarding the therapeutic/toxicity ratio of the treatment. Overcoming her initial refusal involved the use of empathy, knowledge of her perceptions of illness and chemotherapy, education regarding treatment, ample time for information processing, and the involvement of her daughter in decision-making. PMID- 15796441 TI - A multidisciplinary approach to gastrointestinal bleeding in cancer patients. AB - Gastrointestinal bleeding remains one of the most challenging areas in medicine today, especially in the patient with cancer. New techniques and technology have added much to the diagnosis and management of gastrointestinal bleeding. Although gastroenterologists remain at the center of the evaluation of these patients, the incorporation of new techniques has meant that interventional radiologists and nuclear medicine specialists often have important roles in assessing and treating these patients as well. Because patients with cancer are members of a unique group, gastrointestinal bleeding may also require intervention by surgical oncologists, medical oncologists, and radiation oncologists. This review will address the multidisciplinary approach required for the proper diagnosis and management of gastrointestinal bleeding in patients with cancer. PMID- 15796442 TI - The role of radiation therapy in gastrointestinal bleeding. PMID- 15796443 TI - Managing malignancy-associated hyperuricemia with rasburicase. AB - Along with hydration and urinary alkalinization, allopurinol has been the standard agent for the management of hyperuricemia in patients with a high tumor burden who are at risk for tumor lysis syndrome. However, this agent often fails to prevent and treat this complication effectively. Rasburicase, a recombinant urate oxidase, acts at the end of the purine catabolic pathway and, therefore, does not induce accumulation of xanthine or hypoxanthine, which can precipitate in the kidneys and lead to impaired renal function. Rasburicase may represent an effective alternative to allopurinol in rapidly reducing uric acid levels, improving patients' electrolyte status, and reversing renal insufficiency. The drug initially was studied in pediatric patients with acute lymphoblastic leukemia and aggressive non-Hodgkin lymphoma; data may suggest comparable benefit in adults with similar lymphoid malignancies. Current and future trials will evaluate alternate doses and schedules of rasburicase to maintain its efficacy while reducing its cost. PMID- 15796444 TI - Optimizing care with rasburicase. PMID- 15796445 TI - Quality of life outcomes: measurement and intervention. PMID- 15796446 TI - Measuring quality of life in patients with melanoma: development of the FACT melanoma subscale. AB - A systematic review of the literature on quality of life (QOL) in melanoma patients suggested an overwhelming need for a disease-specific subscale. A melanoma subscale for the Functional Assessment of Cancer Therapy (FACT-Melanoma) was developed to meet this need. This instrument was developed in three stages. In stage I, the literature was comprehensively reviewed, and over 300 cancer specific items from the Functional Assessment of Chronic Illness Therapy (FACIT) item bank were examined to identify questions of potential relevance to melanoma patients. In stage II, 20 melanoma experts identified questions that were relevant to melanoma patients and that were to be included in a pilot questionnaire. In stage III, the pilot questionnaire and a semistructured interview to assess item comprehension, relevance, and overall content were administered to 40 patients with various stages of melanoma. In all, 97 items were culled from the literature and the FACIT item bank; after items were reviewed and evaluated, 25 questions were retained. Most patients considered the content of the pilot questionnaire to be relevant (95%), comprehensive (60%), and easy to understand (88%). After final revisions were made, the FACT-Melanoma tool included 24 items encompassing three QOL domains: 20 items relate to physical well-being, 3 to emotional well-being,and 1 to social well-being. The face and content validity of the FACT-Melanoma assessment tool has been confirmed in melanoma patients and by professionals. Formal validation and reliability testing of the questionnaire is being determined in a prospective cohort of melanoma patients. PMID- 15796447 TI - Patient preferences in choosing chemotherapy regimens for advanced non-small cell lung cancer. AB - The major difference between cisplatin-based chemotherapy doublets for advanced non-small cell lung cancer (NSCLC) is not in the outcomes of their use--rather, it is in the side effects and toxicities that they cause. The degree to which oncologists involve lung cancer patients in discussions regarding the selection of chemotherapy is unknown. A questionnaire regarding patient concerns about chemotherapy and physician discussions was sent to patients registered in the Alliance for Lung Cancer Advocacy, Support, and Education (ALCASE) database from 2000--2002. About three-quarters of the respondents reported that if they were given the option, they would consider side effects important in choosing a particular regimen--and nausea was the most important side effect that would influence that decision. Female patients were more likely to worry about infection and hair loss resulting from therapy than were men. Further, about two thirds of patients reported that they had discussed differences in chemotherapy side effects with their physicians, particularly if the physicians were female, although less than half of those patients recalled discussing the selection of a particular regimen based upon its side-effect profile. Different chemotherapy regimens with varied side-effect profiles have been developed, but medical oncologists do not present options for chemotherapy uniformly to their patients based on possible or probable adverse reactions. Better communication between physician and patient about the likelihood of side effects may reduce chemotherapy-related stress for patients. PMID- 15796448 TI - Finding meaning, balance, and personal satisfaction in the practice of oncology. PMID- 15796449 TI - Management of malignant pleural effusions and ascites. PMID- 15796450 TI - Tympanostomy tube otorrhea: treating the first infection. Highlights of a roundtable discussion sponsored by Alcon Laboratories, Inc.; Oct. 16, 2004; Fort Worth, Texas. PMID- 15796451 TI - The gray zone. PMID- 15796452 TI - The Scarlett Letter. PMID- 15796454 TI - The Scarlett Letter. PMID- 15796453 TI - The Scarlett Letter. PMID- 15796455 TI - The Scarlett Letter. PMID- 15796456 TI - How to set up an IRS-approved family loan. PMID- 15796457 TI - Impending crisis in oral health care prompts look at dental education. PMID- 15796458 TI - Analyzing Pennsylvania community residential facilities for individuals with intellectual and developmental disabilities. AB - Evolving residential requirements for individuals with mild and moderate intellectual disabilities and related developmental disabilities increasingly place these persons in community settings. The increasing numbers of these individuals are dependent upon local practitioners for needed health services. National and Pennsylvania data are reviewed in an effort to provide a general awareness of these community living arrangements, which in turn may assist in the delivery and the follow-up of oral health services. PMID- 15796459 TI - [CME-ECG 4. Lost race]. PMID- 15796460 TI - [Improvement of ultrasound education by simulator training]. AB - Ultrasound education is complicated by varying quantities and qualities of specific pathologies resulting from distinct patient collectives. Furthermore, under current clinical conditions ultrasound educators as well as trainees frequently lack the time necessary for a sufficient ultrasound education. Finally, current ultrasound education materials including "gold standard" images are not ideally suited for teaching scanning three-dimensional pathologies. An improvement of ultrasound education is feasible by training ultrasound trainees on an ultrasound simulator we developed. By using this simulator as well in the classical beginner and advanced level courses as in focused courses of abdominal emergencies or general practice for instance, ultrasound trainees are able to practice scanning of numerous well defined pathologies under realistic conditions. PMID- 15796461 TI - [Patient safety in diagnostic ultrasound I--fundamentals]. AB - Diagnostic ultrasound is not totally free of side effects. Therefore, the principle of good clinical practise and ALARA must be considered. In particular inpulsed wave Doppler examinations critical values can be exceeded. Potentially, damage can be the result. Also ultrasound contrast agents are not absolutely free of side effects. The most important risk, of course, are ultrasound examinations performed by inadequat skilled physicians. PMID- 15796462 TI - [Patient safety in diagnostic ultrasound II--ultrasound guided interventions]. AB - New digital ultrasound techniques simplify the documentation of ultrasound images and allow better comparison with respect to different image modalities. New vascular imaging modes including contrast enhanced power Doppler and 3D power Doppler allow for the detection of very small vessels and leakages. B-flow, a recently introduced digital substraction technique is advancing the detection of residual blood flow in high-grade stenosis without vibration artifacts. Modern contrast agents allow to perform ultrasound guided intervetion such as biopsies and ablation therapies with improved accuracy. This way, the risk for complications can be significantly reduced. PMID- 15796463 TI - [Sonography of the adrenal glands]. AB - In the abdominal ultrasonography, the representation of normal adrenal glands is frequently problematic, also for experienced practitioners in ultrasonography. During a seminary at the congress of the SGUM in Davos, in June 2004, it was specially entered to this problematic by anatomical illustrations and live demonstrations. These statements will be summarized in the following article. Also, the technics of examination of the adrenal glands will be explained, especially in comparison to anatomical cut-preparations. It will be entered to particular pathological statements of the adrenal glands. The proceeding will be described by the localisation of accidentally detected tumours of adrenal glands. PMID- 15796464 TI - [Malaise and abdominal complaints in a 24-year-old Swiss-African patient from Guinea: schistosomiasis mansoni]. PMID- 15796465 TI - [CME-Ultrasonography 3/Solution. Acute swallowing pain. Tonsillar abscess]. PMID- 15796466 TI - [A literary office visit]. PMID- 15796467 TI - [Nurses' aide diploma passes validation of acquired experience]. PMID- 15796469 TI - [New deal for handicapped persons]. PMID- 15796468 TI - [Experiments in competence transfers are launched]. PMID- 15796470 TI - [Demographic shock, the health sector in danger?]. PMID- 15796471 TI - [Delegation of acts, the syndicates mobilize]. PMID- 15796472 TI - [11-Clinical aspects: diabetic foot ulcers]. PMID- 15796473 TI - [Nurses during the Resistance]. PMID- 15796474 TI - [Help, mutual aid, power...]. PMID- 15796475 TI - [Cancer, towards a new management]. PMID- 15796476 TI - [Cancer...the France of inequalities]. PMID- 15796477 TI - [The Cancer plan, two years after]. PMID- 15796478 TI - [What psychological support is needed after the cancer diagnosis announcement?]. PMID- 15796479 TI - [Psychoanalysis to the rescue]. PMID- 15796480 TI - [Breast cancer, the nursing consultation in the framework of the Cancer plan]. PMID- 15796482 TI - [Current update on melanoma]. PMID- 15796481 TI - [Cancer, towards a new management. You will tell them...]. PMID- 15796483 TI - [Management of cancer patients, what has changed?]. PMID- 15796484 TI - [Bibliographic elements. Cancer, towards a new management]. PMID- 15796485 TI - [Cardiovascular system drugs. 10/12. The heparins]. PMID- 15796486 TI - [2/6. A consistently prudent driver]. PMID- 15796487 TI - [II-Relationship care. 11. Persons who have difficulty coping with their beliefs and values, cultural and spiritual]. PMID- 15796488 TI - [The short- and long-term results of the "Intensive Care" of 160 sick neonatal foals]. AB - The short- and long-term results of admission to the 'Neonatal Intensive Care Unit' (ICU) at the Department of Equine Sciences of the Utrecht University were evaluated. The patients were categorized, based on specific criteria, into different groups: 'Sepsis', 'Pneumonia', 'Enteritis', 'Asphyxia', 'Premature', 'Isoerythrolysis', and 'Equine herpes virus (EHV)'. The survival rate and mean duration of hospitalization were determined for each group. The venous blood variables at admission were compared for 'surviving' and 'non-surviving' foals within groups, between groups, and for the overall group. From 1992 to 2001, 160 critically ill foals (younger than 7 days) were admitted to the ICU. Thirty-six percent of the foals survived. The mean duration of hospitalization was 6.6 days for the 'surviving' foals and 3.0 days for the 'non-surviving' foals. The pCO2 appeared to the the most important diagnostic and prognostic variable. The results of a telephone questionnaire (owner response of 81 %) revealed that in the long term 60% of the surviving foals lived up to the owner's expectations. PMID- 15796489 TI - [Within the rules]. PMID- 15796490 TI - [General Food Safety]. PMID- 15796491 TI - [Horse Advisory Committee: 'newborn foal']. PMID- 15796492 TI - [Horse Advisory Committee: 'Sperm handling']. PMID- 15796493 TI - [Veterinary antibiotic use: what is different in Denmark?]. PMID- 15796494 TI - [Polycystic Kidney Disease (PKD) in cats]. PMID- 15796495 TI - [Veterinary practice and career symposium]. PMID- 15796496 TI - [Marketing in veterinary practice]. PMID- 15796497 TI - [Instruments, books and other objects memorable to (almost) forgotten opinions, therapies, buildings, et cetera. Sign use in prevention and control of cattle warble infestation]. PMID- 15796498 TI - Design of optical switches as metabolic indicators: new fluorogenic probes for monoamine oxidases (MAO A and B). AB - This study describes the design of sensitive, selective, and fluorogenic reporter substrates for monoamine oxidase (MAO) enzymes. This was achieved by an iterative effort, guided by PET and TICT photophysical concepts, which led to the development of irreversible redox switches based on a facile oxidation cyclization reporting mechanism. Specifically, enzymatic oxidation of the ethylamino group in probe 9 proceeded via a putative aldehyde intermediate, which subsequently underwent spontaneous and intramolecular condensation with the aniline amino group furnishing an indole product in an irreversible fashion. This overall change resulted in a significant change in the emission intensity. When expressed in terms of brightness, the origins of this emission switch may be rationalized by the changes in quantum yield and absorbance strength. The fluorescence readout directly correlated with the kinetics of the oxidative step (i.e., reporting mechanism was fast, the intermediate aldehyde was not detected). Probe 9 is a good substrate for MAO B (Km = 510 +/- 40 muM, kcat = 21 min-1) with the kinetic parameters comparable to physiological substrates. This probe not only allows for direct and continuous measurement of MAO activity in mitochondria and tissue homogenates, but more importantly sets the stage for future studies in intact cells and organs. PMID- 15796501 TI - Reversible O-O bond cleavage and formation of a peroxo moiety of a peroxocarbonate ligand mediated by an iron(III) complex. AB - A mononuclear iron(III) complex containing a peroxocarbonate ligand, [Fe(qn)2(O2C(O)O)]- (qn = quinaldinate), underwent the reversible O-O bond cleavage and reformation of the peroxo group via the formation of FeIV=O or FeV=O species, which was confirmed by the resonance Raman and ESI-TOF/MS measurements. PMID- 15796500 TI - Self-assembly of supramolecular chiral insulated molecular wire. AB - Supramolecular chiral insulated molecular wire was constructed by self-assembly of a neutral one-dimensional schizophyllan host and a water-soluble polythiophene guest. The work presented here will not only open a door to a new application of polysaccharides but also provide an important clue to prepare stable supramolecular insulated molecular wires with one-handed helical structure. PMID- 15796499 TI - Sequence-selective recognition of peptides within the single binding pocket of a self-assembled coordination cage. AB - The single binding pocket of a self-assembled Pd6L4 coordination cage recognizes oligopeptides in a highly sequence-selective fashion. In particular, the Trp-Trp Ala sequence is strongly bound by the cavity (Ka >/=106 M-1). Tripeptides possessing the same residues but in different sequences (i.e., Trp-Ala-Trp and Ala-Trp-Trp) show much poorer affinity. Even singly mutated tripeptides with aromatic-aromatic-aliphatic sequences of the residues (e.g., Trp-Trp-Gly and Trp Tyr-Ala) are not recognized efficiently. X-ray analysis and NMR reveal that all residues of the Trp-Trp-Ala sequence cooperatively interact with the cage via CH pi and pi-pi interactions. PMID- 15796502 TI - Facile C-N cleavage in a series of bridged lactams. AB - A series of strained bi- and tricyclic amides has been shown to be unusually sensitive to cleavage of the C-N bond adjacent to the amide moiety. This bond undergoes facile breaking when subjected to treatment with H2/Pd(OH)2, MeI, and DDQ. In each case, the reaction is highly regioselective and mainly results in breaking the C-N bond that deviates the farthest from its natural planar state. Preliminary experiments that bear on the mechanisms of these reactions are described. PMID- 15796503 TI - Resonance assignment of proteins with high shift degeneracy based on 5D spectral information encoded in G2FT NMR experiments. AB - A suite of novel (5,3)D G2FT triple resonance NMR experiments encoding highly resolved 5D spectral information is presented for sequential resonance assignment of proteins exhibiting high chemical shift degeneracy. Efficient resonance assignment is achieved by separate joint sampling of (i) chemical shifts which solely serve to provide increased resolution and (ii) shifts which also provide sequential connectivities. In these G2FT experiments, two G-matrix transformations are employed. Peaks are resolved along a first GFT dimension at both Omega(15N) + Omega(13C') and Omega(15N) - Omega(13C'), or at Omega(15N) + Omega(13Calpha) and Omega(15N) - Omega(13Calpha), to break backbone 15N,1HN chemical shift degeneracy. Sequential connectivities are established along a second GFT dimension by measuring intraresidue and sequential correlations at 2Omega(13Calpha), Omega(13Calpha + 13Cbeta), and Omega(13Calpha - 13Cbeta), or at Omega(13Calpha + 1Halpha) and Omega(13Calpha - 1Halpha), to resolve 13Calpha/beta,1Halpha chemical shift degeneracy. It is demonstrated that longitudinal proton relaxation optimization of out-and-back implementations suitable for deuterated proteins and nonlinear data sampling combined with maximum entropy reconstruction further accelerate G2FT NMR data acquisition speed. As a result, the spectral information can be obtained within hours, so that (5,3)D G2FT experiments are viable options for high-throughput structure determination in structural genomics. Applications are presented for 17 kDa alpha helical protein YqbG and 13.5 kDa protein rps24e, targets of the Northeast Structural Genomics consortium, as well as for 9 kDa protein Z-domain. The high resolving power of the G2FT NMR experiments makes them attractive choices to study alpha-helical globular/membrane or (partially) unfolded proteins, thus promising to pave the way for NMR-based structural genomics of membrane proteins. PMID- 15796504 TI - Phosphine oxide polymer for water-soluble nanoparticles. AB - A phosphine oxide polymer was developed using bis(dichlorophosphino)ethane and poly(ethylene glycol). This polymer system was used to transfer various nanoparticles from organic solvents to water, retaining their physical properties and reactivities. PMID- 15796506 TI - Competitive decay at two- and three-state conical intersections in excited-state intramolecular proton transfer. AB - We demonstrate the existence of a simultaneous degeneracy (not required by symmetry) of three electronic states in malonaldehyde. This is one of the first reports of such a triple degeneracy involving S0, S1, and S2 in a molecule with a closed-shell ground state. We further report on a two-state S2/S1 conical intersection which is higher in energy than the three-state intersection, but closer to the Franck-Condon point. First-principles quantum dynamics calculations of the photochemistry after excitation to S2 show that there is a competition between these intersections, with more than half of the population decaying to S1 through the higher energy S2/S1 intersection. Surprisingly, much of the population which makes it to the triple degeneracy point is not funneled directly to S0, but rather remains trapped on S1. We attribute this to the large dimensionality of the branching plane at a three-state intersection (the degeneracy is lifted along at least five distinct molecular displacements). PMID- 15796507 TI - Unexpected reactivities of Cu2(diphosphine)2 complexes in alcohol: isolation, x ray crystal structure, and photoluminescent properties of a remarkably stable [Cu3(diphosphine)3(mu3-H)]2+ hydride complex. AB - Treatment of [Cu2(dcpm)2]Y2 (dcpm = bis(dicyclohexylphosphino)methane, Y = ClO4-, BF4-, PF6-, CF3SO3-) with refluxing MeOH in the presence of KOH afforded hydride complexes [Cu3(dcpm)3(mu3-H)]Y2 (1) in about 85% yield. Refluxing [Cu2(dcpm)2](PF6)2 with MeOH in the presence of NH3.H2O and air gave a carboxylate complex [Cu2(dcpm)2(O2CCH2OH)]PF6 (2) in 40% yield. All of the complexes 1 and 2 have been characterized by X-ray crystallography. The Cu3 cores in 1 are almost perfectly shielded by the dcpm ligands. Intense photoluminescence was observed for 1 both in the solid state and in solution. PMID- 15796505 TI - Structures of E. coli peptide deformylase bound to formate: insight into the preference for Fe2+ over Zn2+ as the active site metal. AB - E. coli peptide deformylase (PDF) catalyzes the deformylation of nascent polypeptides generated during protein synthesis. While PDF was originally thought to be a zinc enzyme, subsequent studies revealed that the active site metal is iron. In an attempt to understand this unusual metal preference, high-resolution structures of Fe-, Co-, and Zn-PDF were determined in complex with its deformylation product, formate. In all three structures, the formate ion binds the metal and forms hydrogen-bonding interactions with the backbone nitrogen of Leu91, the amide side chain of Gln50, and the carboxylate side chain of Glu133. One key difference, however, is how the formate binds the metal. In Fe-PDF and Co PDF, formate binds in a bidentate fashion, while in Zn-PDF, it binds in a monodentate fashion. Importantly, these structural results provide the first clues into the origins of PDF's metal-dependent activity differences. On the basis of these structures, we propose that the basis for the higher activity of Fe-PDF stems from the better ability of iron to bind and activate the tetrahedral transition state required for cleavage of the N-terminal formyl group. PMID- 15796508 TI - Patterning phase separation in polymer films with dip-pen nanolithography. AB - We report a rapid-prototyping method for controlling nanoscale phase separation and pattern formation in conjugated polymer blend films using Dip-Pen Nanolithography (DPN). We use DPN to generate patterned alkylthiol monolayers with feature sizes down to 50 nm on gold surfaces and show how such patterns can nucleate the formation of lateral domains in blends of poly-3-hexylthiophene (P3HT) and polystyrene (PS) cast from solution. We show that this process can be used to probe phase nucleation at heterogeneous surface sites ranging in size from 50 to 750 nm, and that polymer features smaller than 150 nm in diameter can be achieved. We anticipate this method will be useful for studying polymer film responses to nanoscale surface fluctuations as well as for correlating nanoscale phase separation with optoelectronic processes in organic films used in light emitting diode and photovoltaic devices. PMID- 15796509 TI - Synthesis and characterization of a coordinated oxoborane: Lewis acid stabilization of a boron-oxygen double bond. AB - The first example of a stable oxoborane monomer (LB=O) stabilized by complexation to AlCl3 has been prepared by the reaction of LAlCl2 with BCl3 followed by treatment with H2O in CH2Cl2 (L = [HC(CMe)2(NC6F5)2]). DFT calculations reveal that considerable boron-oxygen double bond character is retained in this complex. PMID- 15796510 TI - A heterometal single-molecule magnet of [MnIII2NiII2Cl2(salpa)2]. AB - The tetranuclear complex [MnIII2NiII2Cl2(salpa)2] (salpa = N-(2-hydroxybenzyl)-3 amino-1-propanol) has a spin ground state of S = 6 and was confirmed to be an SMM based on a steplike feature of the magnetization hysteresis loop at 0.55 K. PMID- 15796511 TI - Highly extended conformation of polyelectrolytes incorporated into hybrid threadlike micelles studied by small angle neutron scattering. AB - SANS measurements revealed that polyelectrolytes, sodium salt of partially sulfonated polystyrenes, incorporated into enormously long hybrid threadlike micelles formed in aqueous solution with a cationic surfactant, cetyltrimethylammonium bromide, have a highly extended conformation with high confinement along the micelles with a radius of 2.3 nm. PMID- 15796512 TI - Visible light sensitive cyclomer and its tautomeric dispiro compound formed from bispyridinyl diradical. AB - Two-electron reduction of 4,4'-(1,5-pentanediyl)bis(N-methylpyridinium) (52+) with sodium amalgam in degassed acetonitrile leads quantitatively to the formation of a cyclomer of 3,15-dimethyl-3,15-diazatricyclo[10.3.1.12,6]heptadeca 4,6(16),12(17),13-tetraene (6) by intramolecular cyclization. In degassed acetonitrile, 6 is stable for a long time at room temperature and, hence, a highly stable species. In contrast, 6 undergoes a photochemical reaction to give its valence tautomer having a dispiro structure, 3,10-dimethyl-3,10 diazadispiro[5.0.5.5]heptadeca-1,4,8,11-tetraene (7), when irradiated with visible light shorter than 450 nm. Noteworthy is the experimental fact that 7 is unstable to heat and, hence, reverts back to 6 by application of heat. PMID- 15796513 TI - Selective growth of Si nanowire arrays via galvanic displacement processes in water-in-oil microemulsions. AB - Galvanic displacement processes are employed in water-in-oil microemulsions to deposit gold nanoclusters selectively on Si surfaces and sidewalls. The gold clusters then serve as catalysts to achieve selective growth of vertically and laterally aligned Si nanowire arrays by chemical vapor deposition via the vapor liquid-solid growth mechanism. The size of the gold clusters is shown to have a good correlation with the microemulsion parameters, which in turn controls the size of the synthesized nanowires. PMID- 15796514 TI - Role of cholesterol and polyunsaturated chains in lipid-protein interactions: molecular dynamics simulation of rhodopsin in a realistic membrane environment. AB - We present a 118-ns molecular dynamics simulation of rhodopsin embedded in a bilayer composed of a 2:2:1 mixture of 1-stearoyl-2-docosahexaenoyl phosphatidylcholine (SDPC), 1-stearoyl-2-docosahexaenoyl-phosphatidylethanolamine (SDPE), and cholesterol, respectively. The simulation demonstrates that the protein breaks the lateral and transverse symmetry of the bilayer. Lipids near the protein preferentially reorient such that their unsaturated chains interact with the protein, while the distribution of cholesterol in the membrane complements the variations in rhodopsin's transverse profile. The latter phenomenon suggests a molecular-level mechanism for the experimental finding that cholesterol stabilizes the native dark-adapted state of rhodopsin without binding directly to the protein. PMID- 15796515 TI - [5 + 1] annulation: a synthetic strategy for highly substituted phenols and cyclohexenones. AB - A novel [5 + 1] annulation strategy is developed for the synthesis of highly substituted phenols and cyclohexenones from alpha-alkenoyl ketenedithioacetals and nitroalkanes. PMID- 15796516 TI - Pressure-tuning of magnetism and linkage isomerism in iron(II) hexacyanochromate. AB - A pressure-induced linkage isomerization of the cyanide anion has been observed in single crystals of a chromium(III)-iron(II) Prussian blue analogue of formula K0.4Fe4[Cr(CN)6]2.8 square1.2.16H2O (1). Upon application of pressure in the 0 1200 MPa range, the cyanide ligand rotates and becomes C-bonded to the iron(II) cations, leading to a stabilization of their diamagnetic low-spin states. The result is a decrease of magnetization and magnetic ordering temperatures from TC = 19 K at ambient pressure to 13 K at 1200 MPa. The initial magnetic properties can be restored on pressure release. The reversible movement of cyanide in the solid state can be exploited as a switch of the magnetic interaction at the molecular level. PMID- 15796517 TI - Copolymerization of ethylene with cyclohexene (CHE) catalyzed by nonbridged half titanocenes containing aryloxo ligand: notable effect of both cyclopentadienyl and anionic donor ligand for efficient CHE incorporation. AB - Cyclohexene (CHE) has been incorporated into the polymer chain in ethylene/CHE copolymerization by nonbridged half-titanocenes containing aryloxo ligands of the type, Cp'TiCl2(O-2,6-iPr2C6H3), in the presence of methylaluminoxane (MAO) cocatalyst. The effect of the substituent in the cyclopentadienyl fragment was found to be very important for CHE incorporation; both the tert-BuCp (3) and 1,2,4-Me3Cp analogues (4) showed efficient CHE incorporation, whereas a negligible amount of CHE incorporation was observed by both the indenyl (1) and the Cp* analogue (2) under the same conditions. Cp-ketimide analogue, CpTiCl2(N=CtBu2) (5), zirconocene-like Cp2ZrCl2 (6), and linked half-titanocene like [Me2Si(C5Me4)(NtBu)]TiCl2 (7) did not show any CHE incorporation under the same conditions; unique characteristics for using this type of catalyst precursor for the present copolymerization have thus been emphasized. PMID- 15796518 TI - Enantioselective synthesis of nitroalkanes bearing all-carbon quaternary stereogenic centers through Cu-catalyzed asymmetric conjugate additions. AB - The first examples of catalytic asymmetric conjugate addition (ACA) of alkylzinc reagents to trisubstituted nitroalkenes, leading to the formation of nitroalkanes bearing a quaternary carbon stereogenic center, are reported. Reactions are promoted in the presence of 4 mol % of a readily available amino acid-based phosphine and 2 mol % (CuOTf).C6H6. Cu-catalyzed reactions proceed efficiently in up to 98% ee and can be carried out with a variety of dialkylzinc reagents and trisubstituted nitroolefins. We highlight the synthetic utility of the products obtained by efficient conversion of optically enriched nitroalkanes to the corresponding carboxylic acids. PMID- 15796519 TI - Biomimetic synthesis of trans,syn,trans-fused polyoxepanes: remarkable substituent effects on the endo-regioselective oxacyclization of polyepoxides. AB - The biogenesis of trans,syn,trans-fused polycyclic ether substructures of neurotoxic natural products of the brevetoxin-ciguatoxin family may involve regio and stereoselective oxacyclizations of polyepoxides. We report that a biomimetic Lewis acid-promoted cyclization of 1 to 2 proceeds with endo-regioselectivity and anti-stereospecificity at each position of nucleophilic addition (C3, C7, C11, and C15 in 1) when the tandem cyclization cascade is terminated by a carbonyl oxygen nucleophile. In contrast to expectations from earlier reports with simpler epoxide substrates, alkyl substituents are not required at internal epoxide positions corresponding to C7 and C11 in polyepoxide 1. Furthermore, we report that trimethylsilyl serves as a removable regioselectivity-directing substituent at positions corresponding to C3 of 1. These results greatly expand the potential applications of this biomimetic oxacyclization cascade to the synthesis of fused polycyclic ether natural products. PMID- 15796520 TI - Laser-induced fragmentation of transition metal nanoparticles in ionic liquids. AB - Stable Pd(0) and Rh(0) nanoparticles with small and narrow size distribution can be prepared from relative large and agglomerated transition-metal particles dispersed in 1-n-butyl-3methylimidazolium hexafluorophosphate ionic liquid by simple laser irradiation. The laser irradiation is a complementary method for the generation of stable metal colloids in ionic liquids and also for the regeneration of small-size nanoparticles that may result from their agglomeration after different applications. PMID- 15796521 TI - Allenyl azide cycloaddition chemistry. synthesis of pyrrolidine-containing bicycles and tricycles via the possible intermediacy of azatrimethylenemethane species. AB - Thermolysis of 5-azidoallenes bearing a C(1) methyl group and either an aryl ring or an alkene on C(1) furnishes tricyclic (from the aryl substrates) or bicyclic (from the alkenyl substrates) pyrrolidine products following formal H-CN addition across an intermediate imine. High levels of diastereoselectivity are observed in all cases studied. This reaction cascade presumably passes through unobserved triazoline and azatrimethylenemethane diyl intermediates en route to product. PMID- 15796522 TI - Pd-catalyzed C3-selective allylation of indoles with allyl alcohols promoted by triethylborane. AB - Under palladium catalysis, Et3B nicely promotes allyl alcohols to undergo C3 selective allylation of indoles and tryptophan; the yields range 75-95%. PMID- 15796523 TI - Asymmetric nickel-catalyzed Negishi cross-couplings of secondary alpha-bromo amides with organozinc reagents. AB - A Ni/Pybox catalyst achieves the asymmetric cross-coupling of secondary alpha bromo amides with organozinc reagents. The process tolerates a variety of functional groups and affords the desired product in good yield and in high enantiomeric excess. PMID- 15796524 TI - Total synthesis of cribrostatin IV: fine-tuning the character of an amide bond by remote control. AB - We report the enantioselective total synthesis of cribrostatin IV (1). Key features of this synthesis involve the convergent coupling of two highly functionalized homochiral components followed by a "lynchpin" Mannich cyclization to establish the pentacyclic core (cf. 19 --> 20). PMID- 15796526 TI - Methyl scanning: total synthesis of demethylasterriquinone B1 and derivatives for identification of sites of interaction with and isolation of its receptor(s). AB - The principle of methyl scanning is proposed for determination of the sites of interaction between biologically active small molecules and their macromolecular target(s). It involves the systematic preparation of a family of methylated derivatives of a compound and their biological testing. As a functional assay, the method can identify the regions of a molecule that are important (and unimportant) for biological activity against even unknown targets, and thus provides an excellent complement to structural biology. Methyl scanning was applied to demethylasterriquinone B1, a small-molecule mimetic of insulin. A new, optimal total synthesis of this natural product was developed that enables the family of methyl scan derivatives to be concisely prepared for evaluation in a cellular assay. The results of this experiment were used to design a biotin demethylasterriquinone conjugate for use as an affinity reagent. This compound was prepared in tens of milligram quantities in a four-step synthesis. PMID- 15796525 TI - Hammett analysis of selective thyroid hormone receptor modulators reveals structural and electronic requirements for hormone antagonists. AB - Selective thyroid hormone modulators that function as isoform-selective agonists or antagonists of the thyroid hormone receptors (TRs) might be therapeutically useful in diseases associated with aberrant hormone signaling. The most potent thyroid hormone antagonist reported to date is NH-3. To explore the significance of the 5'-p-nitroaryl moiety of NH-3 and understand what chemical features are important to confer antagonism, we sought to expand the structure-activity relationship data for the class of 5'-phenylethynyl GC-1 derivatives. Herein, we describe an improved synthetic route utilizing palladium-catalyzed chemistry for efficient access to a series of 5'-phenylethynyl compounds with varying size and electronic properties. We prepared and tested sixteen analogues for TR binding and transactivation activity. Substitution at the 5'-position decreased binding affinity, but retained TRbeta-selectivity. In transactivation assays, the analogues displayed a spectrum of agonist, antagonist, and mixed agonist/antagonist activity that correlated with electronic character in a Hammett analysis between sigma substituent value and TR modulation. Analogues NH 5, NH-7, NH-9, NH-11, and NH-23 displayed full antagonist activity with reduced potency compared to NH-3, indicating the nitro group is not required for antagonism. However, para-substitution with strong electron withdrawing properties on the 5'-aryl extension is important for antagonist activity, and antagonist potency-but not ligand receptor binding-was found to correlate linearly with the sigma values for the electron withdrawing substituents. PMID- 15796527 TI - Resonance Raman and infrared spectroscopic studies of high-output forms of human soluble guanylyl cyclase. AB - The allosteric regulator BAY-41-2272 converts the CO adduct of soluble guanylyl cyclase (CO-sGC) enzyme from a low- to high-output form, with respect to production of cGMP. Resonance Raman (RR) and Fourier Transform Infrared (FTIR) spectroscopic techniques are used to show that the CO-sGC exists as major and minor conformers, both having nu(Fe-CO) and nu(C-O) modes characteristic of 6 coordinate species. It is further shown that addition of BAY-41-2272 to the CO adduct induces the transition of some fraction of the initial CO-heme adducts into two new CO-heme complexes, the fractional conversion being dependent on the temperature. One new complex displays vibrational modes characteristic of pentacoordinated CO-adduct, and its formation is not affected by temperature. The second complex, although slightly different from the original CO-adducts, is hexacoordinated, and its formation is facilitated by temperature. The production of substantial amounts of the 5-coordinate CO adduct upon addition of BAY-41 2272, reveals the fact that several out-of-plane heme deformation modes are simultaneously activated, an observation similar to that realized upon NO activation. While the precise nature of these modes will require elucidation by isotopic labeling experiments, by analogy with earlier studies of other heme proteins, several bands associated with modes attributable to peripheral substituent deformations and methine carbon movements are implicated. The documented formation of two new forms upon addition of Bay-41-2272 (a 5 coordinate and a new 6-coordinate form) is discussed with respect to the implications for enzyme activation. PMID- 15796528 TI - Structure-based optimization of a non-beta-lactam lead results in inhibitors that do not up-regulate beta-lactamase expression in cell culture. AB - Bacterial expression of beta-lactamases is the most widespread resistance mechanism to beta-lactam antibiotics, such as penicillins and cephalosporins. There is a pressing need for novel, non-beta-lactam inhibitors of these enzymes. One previously discovered novel inhibitor of the beta-lactamase AmpC, compound 1, has several favorable properties: it is chemically dissimilar to beta-lactams and is a noncovalent, competitive inhibitor of the enzyme. However, at 26 microM its activity is modest. Using the X-ray structure of the AmpC/1 complex as a template, 14 analogues were designed and synthesized. The most active of these, compound 10, had a K(i) of 1 microM, 26-fold better than the lead. To understand the origins of this improved activity, the structures of AmpC in complex with compound 10 and an analogue, compound 11, were determined by X-ray crystallography to 1.97 and 1.96 A, respectively. Compound 10 was active in cell culture, reversing resistance to the third generation cephalosporin ceftazidime in bacterial pathogens expressing AmpC. In contrast to beta-lactam-based inhibitors clavulanate and cefoxitin, compound 10 did not up-regulate beta lactamase expression in cell culture but simply inhibited the enzyme expressed by the resistant bacteria. Its escape from this resistance mechanism derives from its dissimilarity to beta-lactam antibiotics. PMID- 15796529 TI - Unexpected relative aqueous solubilities of a phosphotyrosine analogue and two phosphonate derivatives. AB - Phosphotyrosine (pTyr) is an essential component of biological signaling, often being a determinant of protein-protein interactions. Accordingly, a number of drug discovery efforts targeting signal transduction pathways have included phosphotyrosine and analogues as essential components of the lead compounds. Toward the goal of improved biological efficacy, the phosphonate and difluoro phosphonate analogues of pTyr have been employed in inhibitor design because of their stability to hydrolysis and enhanced binding affinity in certain cases. To quantitate the contribution of aqueous solubility of pTyr, phosphonomethyl phenylalanine (Pmp), and difluorophosphonomethyl phenylalanine (F(2)Pmp) to their relative binding affinities, free energy perturbation calculations were undertaken on the mimetics phenol phosphate (PP), benzyl phosphonate (BP), and difluorobenzyl phosphonate (F(2)BP), including development of empirical force field parameters compatible with the CHARMM all-atom force fields. Notably, it is shown that the most favorably solvated compound of the series is BP, followed by PP, with F(2)BP the least favorably solvated for both the mono- and dianionic forms of the compounds. The molecular origin of this ordering is shown to be due to changes in charge distribution, in the comparatively larger size of the fluorine atoms, as well as in differences of local solvation between PP and BP. The implications of the differences in aqueous solubility toward the relative binding potencies of pTyr-, Pmp-, and F(2)Pmp-containing peptide ligands are discussed. Our results indicate that one general principle explaining the efficacy of selective fluorination to enhance binding affinities may lie in the ability of fluorine atoms to increase the hydrophobicity of a ligand while maintaining its capability to form hydrogen bonds. PMID- 15796530 TI - Binding mode and transcriptional activation potential of high affinity ligands for the CBP KIX domain. AB - We recently described a pair of ligands, PPKID4(P) (4(P)) and PPKID6(U) (6(U)), which present the alpha-helical functional epitope found on helix B of the CREB KID activation domain (KID(P)) on a pancreatic fold protein scaffold. 4(P) and 6(U) bind the natural target of KID(P), the KIX domain of the coactivator CBP, with equilibrium dissociation constants between 515 nM and 1.5 microM and compete effectively with KID(P) for binding to CBP KIX (KIX). Here we present a detailed investigation of the binding mode, orientation, and transcriptional activation potential of 4(P) and 6(U). Equilibrium binding experiments using a panel of well characterized KIX variants support a model in which 4(P) binds KIX in a manner that closely resembles that of KID(P) but 6(U) binds an overlapping, yet distinct region of the protein. Equilibrium binding experiments using a judiciously chosen panel of 4(P) variants containing alanine or sarcosine substitutions along the putative alpha- or PPII helix of 4(P) support a model in which 4(P) folds into a pancreatic fold structure upon binding to KIX. Transcriptional activation assays performed in HEK293 cells using GAL4 DNA-binding domain fusion proteins indicate that 4(P) functions as a potent activator of p300/CBP-dependent transcription. Notably, 6(U) is a less potent transcriptional activator in this context than 4(P)despite the similarity of their affinities for CBP KIX. This final result suggests that thermodynamic affinity is an important, although not exclusive, criterion controlling the level of KIX-dependent transcriptional activation. PMID- 15796531 TI - RNA SHAPE chemistry reveals nonhierarchical interactions dominate equilibrium structural transitions in tRNA(Asp) transcripts. AB - Current models assume that RNA folding is strongly hierarchical such that the base-paired secondary structure is more stable than and forms independently of the tertiary structure. This model has been difficult to test due to the experimental inability to interrogate the local environment at every nucleotide as a comprehensive function of the RNA folding state. Reaction of an RNA 2' hydroxyl group with N-methylisatoic anhydride to form a nucleotide 2'-ester is governed by the extent to which the nucleotide forms base pairing or tertiary interactions. Selective 2'-Hydroxyl Acylation analyzed by Primer Extension (SHAPE) is shown to be an RNA analogue of the protein hydrogen exchange experiment. Single nucleotide resolution SHAPE analysis emphasizes a complexity for the unfolding of tRNA(Asp) transcripts that is not anticipated by current models for RNA folding. We quantify six well-defined transitions for tRNA(Asp) transcripts between 35 and >75 degrees C, including asymmetric unfolding of the two strands in a single helix, multistep loss of tertiary interactions, and a multihelix conformational shift. The three lowest temperature transitions each involve coupled interactions between the secondary and tertiary structure. Thus, even for this simple RNA, multiple nonhierarchical and complex interactions dominate the equilibrium transitions most accessible from the native state. PMID- 15796532 TI - Tryptophan side chain electrostatic interactions determine edge-to-face vs parallel-displaced tryptophan side chain geometries in the designed beta-hairpin "trpzip2". AB - The interaction geometries of the four tryptophan (Trp) side chains in the 12 residue designed beta-hairpin trpzip2 are investigated using all-atom explicit solvent molecular dynamics simulations. The experimentally observed edge-to-face (EtF) pairwise interaction geometries are stable on a time scale of 10 ns. However, removing the electrostatic multipoles of the Trp side chains while retaining the dipoles of the side chains' NH moieties induces a conformational change to a geometry in which three of the four side chains interact in a parallel-displaced (PD) manner. Free energy simulations of the Etf to PD conformational change reveal that, with the side chain multipole moments intact (+MP), the EtF conformation is preferred by 5.79 kcal/mol. Conversely, with only the dipole moments of the side chain NH moieties intact (-MP), the PD conformation's free energy is more favorable by 1.71 kcal/mol. In contrast to energetic similarities for Trp side chain-water electrostatic and Trp side chain Trp side chain and Trp side chain-water van der Waals, +MP Trp side chain-Trp side chain electrostatic interactions are more favorable by 4.21 kcal/mol in the EtF conformation, while in the -MP case the EtF and PD conformations' Trp side chain-Trp side chain electrostatic energies are nearly identical. The results highlight the importance of electrostatic multipole moments in determining aromatic-aromatic interaction geometries in aqueous biomolecular systems and argue for the inclusion of this physics in simplified models used for protein ligand docking and protein structure prediction, possibly through a truncated Coulomb term between aromatic moieties. PMID- 15796533 TI - Entropic stabilization of isolated beta-sheets. AB - Temperature-dependent electric deflection measurements have been performed for a series of unsolvated alanine-based peptides (Ac-WA(n)-NH(2), where Ac = acetyl, W = tryptophan, A = alanine, and n = 3, 5, 10, 13, and 15). The measurements are interpreted using Monte Carlo simulations performed with a parallel tempering algorithm. Despite alanine's high helix propensity in solution, the results suggest that unsolvated Ac-WA(n)-NH(2) peptides with n > 10 adopt beta-sheet conformations at room temperature. Previous studies have shown that protonated alanine-based peptides adopt helical or globular conformations in the gas phase, depending on the location of the charge. Thus, the charge more than anything else controls the structure. PMID- 15796534 TI - Discrimination of nitroxyl and nitric oxide by water-soluble Mn(III) porphyrins. AB - The water-soluble manganese(III) meso-tetrakis (N-ethylpyridinium-2-yl) porphyrin (Mn(III)TEPyP) and manganese(III) meso-(tetrakis(4-sulfonato-phenyl)) porphyrinate (Mn(III)TPPS) are able to chemically distinguish between HNO and NO donors, reacting with the former in a fast, efficient, and selective manner with concomitant formation of the {MnNO}(7) complex (k(on(HNO)) approximately equal to 10(5) M(-1) s(-1)), while they are inert or react very slowly with NO donors. DFT calculations and kinetic data suggest that HNO trapping is operative at least in the case of Mn(III)TPPS, while catalytic decomposition of the HNO donors (sodium trioxodinitrate and toluene sulfohydroxamic acid) seems to be the main pathway for Mn(III)TEPyP. In the presence of oxygen, the product Mn(II)TEPyP(NO) oxidizes back to Mn(III)TEPyP, making it possible to process large ratios of nitroxyl donor with small amounts of porphyrin. PMID- 15796535 TI - Catalysts for Suzuki-Miyaura coupling processes: scope and studies of the effect of ligand structure. AB - Suzuki-Miyaura coupling reactions of aryl and heteroaryl halides with aryl-, heteroaryl- and vinylboronic acids proceed in very good to excellent yield with the use of 2-(2',6'-dimethoxybiphenyl)dicyclohexylphosphine, SPhos (1). This ligand confers unprecedented activity for these processes, allowing reactions to be performed at low catalyst levels, to prepare extremely hindered biaryls and to be carried out, in general, for reactions of aryl chlorides at room temperature. Additionally, structural studies of various 1.Pd complexes are presented along with computational data that help elucidate the efficacy that 1 imparts on Suzuki Miyaura coupling processes. Moreover, a comparison of the reactions with 1 and with 2-(2',4',6'-triisopropylbiphenyl)diphenylphosphine (2) is presented that is informative in determining the relative importance of ligand bulk and electron donating ability in the high activity of catalysts derived from ligands of this type. Further, when the aryl bromide becomes too hindered, an interesting C-H bond functionalization-cross-coupling sequence intervenes to provide product in high yield. PMID- 15796536 TI - Thermodynamic and kinetic control in selective ligand transfer in conjugate addition of mixed organocuprate Me(X)CuLi. AB - Since the proposal of the dummy ligand concept by Corey, it has been widely accepted that the ligand transfer selectivity of a mixed organocuprate (Me(X)CuLi) depends on the Cu-X bond strength. The present B3LYP density functional studies on the Me(2)(X)Cu(III).OMe(2), pi-allyl Cu(III), and Me(X)Cu(III)LiCl.LiCl reacting with acrolein showed that the ligand transfer selectivity of the conjugate addition depends on two factors, thermodynamic stability (X = tert-butyl, ethynyl, cyano, and thiomethyl groups) and kinetic reactivity ((trimethylsilyl)methyl and vinyl groups) of the Cu(III) intermediate formed by complexation of the cuprate and the alpha,beta-unsaturated carbonyl compound. For the typical dummy ligands (X = alkynyl, cyano, and heteroatom ligands), the trans effect and the strong Li-X affinity are the reasons why these ligands stay on the copper atom. In contrast, for the (trimethylsilyl)methyl and vinyl groups, the selectivity depends on the kinetics of reductive elimination of the Cu(III) intermediate. The (trimethylsilyl)methyl transfer is retarded by repulsive four-electron interaction between the lone pair Cu 3d(xy)() orbital and the C-Si sigma-orbital. PMID- 15796537 TI - Eliminating positively charged lysine epsilon-NH3+ groups on the surface of carbonic anhydrase has no significant influence on its folding from sodium dodecyl sulfate. AB - This study compares the folding of two polypeptides--bovine carbonic anhydrase (BCA) and peracetylated BCA (BCA-Ac(18))--having the same sequence of amino acids but differing by 18 formal units of charge, from a solution containing denaturing concentrations of sodium dodecyl sulfate (SDS). Acetylation of BCA with acetic anhydride converts all 18 lysine-epsilon-NH(3)(+) groups to lysine-epsilon NHCOCH(3) groups and generates BCA-Ac(18). Both BCA and BCA-Ac(18) are catalytically active, and circular dichroism spectroscopy (CD) suggests that they have similar secondary and tertiary structures. SDS at concentrations above approximately 10 mM denatured both proteins. When the SDS was removed by dialysis, both proteins were regenerated in native form. This study suggests that large differences in the net charge of the polypeptide have no significant influence on the structure, the ability to refold, or the rate of refolding of this protein from solutions containing SDS. This study reinforces the idea that charged residues on the surface of BCA do not guide protein folding and raises the broader question of why proteins have charged residues on their surface, outside of the region of the active site. PMID- 15796538 TI - Characterization of the FKBP.rapamycin.FRB ternary complex. AB - Rapamycin is an important immunosuppressant, a possible anticancer therapeutic, and a widely used research tool. Essential to its various functions is its ability to bind simultaneously to two different proteins, FKBP and mTOR. Despite its widespread use, a thorough analysis of the interactions between FKBP, rapamycin, and the rapamycin-binding domain of mTOR, FRB, is lacking. To probe the affinities involved in the formation of the FKBP.rapamycin.FRB complex, we used fluorescence polarization, surface plasmon resonance, and NMR spectroscopy. Analysis of the data shows that rapamycin binds to FRB with moderate affinity (K(d) = 26 +/- 0.8 microM). The FKBP12.rapamycin complex, however, binds to FRB 2000-fold more tightly (K(d) = 12 +/- 0.8 nM) than rapamycin alone. No interaction between FKBP and FRB was detected in the absence of rapamycin. These studies suggest that rapamycin's ability to bind to FRB, and by extension to mTOR, in the absence of FKBP is of little consequence under physiological conditions. Furthermore, protein-protein interactions at the FKBP12-FRB interface play a role in the stability of the ternary complex. PMID- 15796539 TI - Elucidating the vibrational spectra of hydrogen-bonded aggregates in solution: electronic structure calculations with implicit solvent and first-principles molecular dynamics simulations with explicit solvent for 1-hexanol in n-hexane. AB - Fourier transform infrared spectroscopy is a popular method for the experimental investigation of hydrogen-bonded aggregates, but linking spectral information to microscopic information on aggregate size distribution and aggregate architecture is an arduous task. Static electronic structure calculations with an implicit solvent model, Car-Parrinello molecular dynamics (CPMD) using the Becke-Lee-Yang Parr (BLYP) exchange and correlation energy functionals and classical molecular dynamics simulations for the all-atom version of the optimized parameters for liquid simulations (OPLS-AA) force field were carried out for an ensemble of 1 hexanol aggregates solvated in n-hexane. The initial configurations for these calculations were size-selected from a distribution of aggregates obtained from a large-scale Monte Carlo simulation. The vibrational spectra computed from the static electronic structure calculations for monomers and dimers and from the CPMD simulations for aggregates up to pentamers demonstrate the extent of the contribution of dangling or nondonating hydroxyl groups found in linear and branched aggregates to the "monomeric" peak. Furthermore, the computed spectra show that there is no simple relationship between peak shift and aggregate size nor architecture, but the effect of hydrogen-bond cooperativity is shown to differentiate polymer-like (cooperative) and dimer-like (noncooperative) hydrogen bonds in the vibrational spectrum. In contrast to the static electronic structure calculations and the CPMD simulations, the classical molecular dynamics simulations greatly underestimate the vibrational peak shift due to hydrogen bonding. PMID- 15796540 TI - Multielectron redox chemistry of iron porphyrinogens. AB - Iron octamethylporphyrinogens were prepared and structurally characterized in three different oxidation states in the absence of axial ligands and with sodium or tetrafluoroborate as the only counterions. Under these conditions, the iron- and ligand-based redox chemistry of iron porphyrinogens can be defined. The iron center is easily oxidized by a single electron (E(1/2) = -0.57 V vs NHE in CH(3)CN) when confined within the fully reduced macrocycle. The porphyrinogen ligand also undergoes oxidation but in a single four-electron step (E(p) = +0.77 V vs NHE in CH(3)CN); one of the ligand-based electrons is intercepted for the reduction of Fe(III) to Fe(II) to result in an overall three-electron oxidation process. The oxidation equivalents in the macrocycle are stored in C(alpha) C(alpha) bonds of spirocyclopropane rings, formed between adjacent pyrroles. EPR, magnetic and Mossbauer measurements, and DFT computations of the redox states of the iron porphyrinogens reveal that the reduced ligand gives rise to iron in intermediate spin states, whereas the fully oxidized ligand possesses a weaker sigma-donor framework, giving rise to high-spin iron. Taken together, the results reported herein establish a metal-macrocycle cooperativity that engenders a multielectron chemistry for iron porphyrinogens that is unavailable to heme cofactors. PMID- 15796541 TI - Exciton migration in rigid-rod conjugated polymers: an improved Forster model. AB - The dynamics of interchain and intrachain excitation energy transfer taking place in a polyindenofluorene endcapped with perylene derivatives is explored by means of ultrafast spectroscopy combined with correlated quantum-chemical calculations. The experimental data indicate faster exciton migration in films with respect to solution as a result of the emergence of efficient channels involving hopping between chains in close contact. These findings are supported by theoretical simulations based on an improved Forster model. Within this model, the rates are expressed according to the Fermi golden rule on the basis of (i) electronic couplings that take account of the detailed shape of the excited-state wave functions (through the use of a multicentric monopole expansion) and (ii) spectral overlap factors computed from the simulated acceptor absorption and donor emission spectra with explicit coupling to vibrations (considered within a displaced harmonic oscillator model); inhomogeneity is taken into account by assuming a distribution of chromophores with different conjugation lengths. The calculations predict faster intermolecular energy transfer as a result of larger electronic matrix elements and suggest a two-step mechanism for intrachain energy transfer with exciton hopping along the polymer backbone as the limiting step. Injecting the calculated hopping rates into a set of master equations allows the modeling of the dynamics of exciton transport along the polyindenofluorene chains and yields ensemble-averaged energy-transfer rates in good agreement with experiment. PMID- 15796542 TI - Ruthenium-catalyzed cycloisomerizations of diynols. AB - A wide variety of diynols containing tertiary, secondary, and primary propargylic alcohols undergo a cycloisomerization reaction to form dienones and dienals in the presence of a catalytic amount of [CpRu(CH(3)CN)(3)]PF(6). The formation of five- and six-membered rings is possible using this methodology. Secondary diynols react to form single geometrical isomeric dienones and -als. Primary diynols undergo a cycloisomerization as well as a hydrative cyclization process. The utility of primary diynol cycloisomerization is demonstrated in a synthesis of (+)-alpha-kainic acid. PMID- 15796544 TI - Highly efficient photoinitiation in the cerium(III)-catalyzed aqueous autoxidation of sulfur(IV). An example of comprehensive evaluation of photoinduced chain reactions. AB - The photoinitiated and cerium(III)-catalyzed aqueous reaction between sulfite ion and oxygen has been studied in a diode-array spectrophotometer using the same light beam for excitation and detection. Cerium(III) is identified as the photoactive absorbing species, and the production of cerium(IV) initiates a radical chain reaction. To interpret all the experimental findings, a simple scheme is proposed, in which the additional chain carriers are sulfite ion radical (SO3(-.)), sulfate ion radical (SO4(-.), and peroxomonosulfate ion radical (SO5(-.). The overall rate of oxidation is proportional to the square root of the light intensity per unit volume, which is readily interpreted by the second-order termination reaction of the proposed scheme. It is also shown that the reaction proceeds for an extended period of time in the dark following illumination, and a quantitative analysis is presented for this phase as well. The postulated model predicts that cerium(III) should have a cocatalytic or synergistic effect on the autoxidation of sulfite ion in the presence of other catalysts. This prediction was confirmed in the iron(III)-sulfite ion-oxygen system. The experimental method and the mathematical treatment used might be applicable to a wide range of photoinduced chain reactions. PMID- 15796543 TI - A dopaquinone model that mimics the water addition step of cofactor biogenesis in copper amine oxidases. AB - The consensus mechanism for biogenesis of the 2,4,5-trihydroxyphenylalanine quinone (TPQ) cofactor in copper amine oxidases involves a key water addition to the dopaquinone intermediate. Although hydration of o-quinones seems straightforward and was implicated previously in aqueous autoxidation of catechols to give ultimately hydroxyquinones, a recent study (Mandal, S.; Lee, Y.; Purdy, M. M.; Sayre, L. M. J. Am. Chem. Soc. 2000, 122, 3574-3584) showed that the observed hydroxyquinones arise not from hydration, but from addition to the o-quinones of H(2)O(2) generated during autoxidation of the catechols. In the enzyme case, hydration of dopaquinone is proposed to be mediated by the active site Cu(II). To establish precedent for this mechanism, we engineered a catechol tethered to a Cu(II)-coordinating unit, such that the corresponding o-quinone could be generated in situ by oxidation with periodate (to avoid generation of H(2)O(2)). Thus, coordination of 4-((2-(bis(2 pyridylmethyl)amino)ethylamino)methyl)-1,2-benzenediol (1) to Cu(II) and subsequent addition of periodate resulted in rapid formation of the TPQ-like corresponding hydroxyquinone. Hydroxyquinone formation was seen also using Zn(II) and Ni(II), but not in the absence of M(II). Under the same conditions, periodate oxidation of the simple catechol 4-tert-butylcatechol does not give hydroxyquinone in the presence or absence of Cu(II). M(II)OH(2) pK(a) data for the Cu(II), Zn(II), and Ni(II) complexes with the pendant tetradentate ligand in the masked (dimethyl ether) catechol form, and kinetic pH-rate profiles of the metal-dependent hydroxyquinone formation from periodate oxidation of catechol 1, suggested a rate-limiting addition step of the ligand-coordinated M(II)OH to the o-quinone intermediate. This study represents the first chemical demonstration of a true o-quinone hydration, which occurs in cofactor biogenesis in copper amine oxidases. PMID- 15796545 TI - Synthesis and reactivity of dimeric Ar'TlTlAr' and trimeric (Ar"T1)3 (Ar', Ar" = bulky terphenyl group) thallium(I) derivatives: Tl(I)-Tl(I) bonding in species ligated by monodentate ligands. AB - The synthesis and characterization of three new organothallium(I) compounds are reported. Reaction of (Ar'Li)(2) (Ar' = C(6)H(3)-2,6-(C(6)H(3)-2,6-Pr(i)(2))(2)) and Ar"Li (Ar" = C(6)H(3)-2,6-(C(6)H(3)-2,6-Me(2))(2)) with TlCl in Et(2)O afforded (Ar'Tl)(2) (1) and (Ar' 'Tl)(3) (2). The "dithallene" 1 is the heaviest group 13 dimetallene and features a planar, trans-bent structure with Ar'Tl-Tl = 119.74(14) degrees and Tl-Tl = 3.0936(8) A. Compound 2 is the first structurally characterized neutral, three-membered ring species of formula c-(MR)(3) (M = Al Tl; R = organo group). The Tl(3) ring has Tl-Tl distances in the range ca. 3.21 3.37 A as well as pyramidal Tl geometries. The Tl-Tl bonds in 1 and 2 are outside the range (2.88-2.97 A) of Tl-Tl single bonds in R(2)TlTlR(2) compounds. The weak Tl-Tl bonding in 1 and 2 leads to their dissociation into Ar'Tl and Ar' 'Tl monomers in hexane. The Ar'Tl monomer behaves as a Lewis base and readily forms a 1:1 donor-acceptor complex with B(C(6)F(5))(3) to give Ar'TlB(C(6)F(5))(3), 3. Adduct 3 features an almost linear thallium C(ipso)-Tl-B angle of 174.358(7) degrees and a Tl-B distance of 2.311(2) A, which indicates strong association. Treatment of 1 with a variety of reagents resulted in no reactions. The lower reactivity of 1 is in accord with the reluctance of Tl(I) to undergo oxidation to Tl(III) due to the unreactive character of the 6s(2) electrons. PMID- 15796546 TI - Iridium(0) nanocluster, acid-assisted catalysis of neat acetone hydrogenation at room temperature: exceptional activity, catalyst lifetime, and selectivity at complete conversion. AB - Acetone hydrogenation catalysis is important in applications such as heat pumps and fuel cells or in fulfilling the sizable demand for the product of selective acetone hydrogenation, 2-propanol. Reported herein is the discovery of a superior acetone hydrogenation catalyst--superior in terms of activity at low temperature, selectivity at complete conversion, and total catalyst lifetime. The new catalyst system consists of Ir(0)(n) nanoclusters plus HCl easily and reproducibly formed from commercially available [(1,5-COD)IrCl](2) under H(2). The resultant, room temperature, high activity, and highly selective (2/n)Ir(0)(n) plus 2HCl catalyst system hydrogenates acetone at 22 degrees C and 40 psig of H(2) pressure to 95% 2 propanol and the rest diisopropyl ether at 100% conversion with 16400 total catalytic turnovers and with an initial turnover frequency of 1.9 s(-1) at 22 degrees C. When molecular sieves are added, the catalyst system becomes even more selective and long-lived, providing the complete and selective conversion of acetone to 100% 2-propanol with 188000 total turnovers. Also reported are initial kinetic, D-labeling and other mechanistic studies, a summary section detailing the four main findings, the "green chemistry" aspects, and the current main drawback (a limited catalytic lifetime due to nanocluster precipitation) of the present invention. A review of the extensive literature of acetone hydrogenation is also tabulated as part of the Supporting Information. PMID- 15796547 TI - Thermodynamics, kinetics, and mechanism of (silox)3M(olefin) to (silox)3M(alkylidene) rearrangements (silox = tBu3SiO; M = Nb, Ta). AB - Olefin complexes (silox)(3)M(ole) (silox = (t)Bu(3)SiO; M = Nb (1-ole), Ta (2 ole); ole = C(2)H(4), C(2)H(3)Me, C(2)H(3)Et, C(2)H(3)C(6)H(4)-p-X (X = OMe, H, CF(3)), C(2)H(3)(t)Bu, (c)C(5)H(8), (c)C(6)H(10), (c)C(7)H(10) (norbornene)) rearrange to alkylidene isomers (silox)(3)M(alk) (M = Nb (1=alk), Ta (2=alk); alk = CHMe, CHEt, CH(n)Pr, CHCH(2)C(6)H(4)-p-X (X = OMe, H, CF(3) (Ta only)), CHCH(2)(t)Bu, (c)C(5)H(8), (c)C(6)H(10), (c)C(7)H(10) (norbornylidene)). Kinetics and labeling experiments suggest that the rearrangement proceeds via a delta abstraction on a silox CH bond by the beta-olefin carbon to give (silox)(2)RM(kappa(2)-O,C-OSi(t)Bu(2)CMe(2)CH(2)) (M = Nb (4-R), Ta (6-R); R = Me, Et, (n)Pr, (n)Bu, CH(2)CH(2)C(6)H(4)-p-X (X = OMe, H, CF(3) (Ta only)), CH(2)CH(2)(t)Bu, (c)C(5)H(9), (c)C(6)H(11), (c)C(7)H(11) (norbornyl)). A subsequent alpha-abstraction by the cylometalated "arm" of the intermediate on an alpha-CH bond of R generates the alkylidene 1=alk or 2=alk. Equilibrations of 1 ole with ole' to give 1-ole' and ole, and relevant calculations on 1-ole and 2 ole, permit interpretation of all relative ground and transition state energies for the complexes of either metal. PMID- 15796548 TI - Cooperative metal-boron interactions in the reaction of nido-1,2-(Cp*RuH)2B3H7, Cp* = eta5-C5Me5, with HC(triple bond)CPh. AB - Products of the reaction of nido-1,2-(CpRuH)(2)B(3)H(7), 1, and phenylacetylene demonstrate the ways in which cluster metal and main group fragments can combine with an alkyne. Observed at 22 degrees C are (a) reduction to mu-alkylidene Ru-B bridges (isomers nido-1,2-(CpRu)(2)(1,5-mu-C{Ph}Me)B(3)H(7), 2, and nido-1,2 (CpRu)(2)(1,5-mu-C{CH(2)Ph}H)B(3)H(7), 3), (b) reduction to exo-cluster alkyl substituents on boron (nido-1,2-(CpRuH)(2)-3-CH(2)CH(2)Ph-B(3)H(6), 4), (c) cluster insertion with extrusion of a BH(2) fragment into an exo-cluster bridge (nido-1,2-(CpRu)(2)(mu-H)(mu-BH(2))-4-or-5-Ph-4,5-C(2)B(2)H(5), 5), (d) combined insertion with BH(2) extrusion and reduction (nido-1,2-(CpRu)(2)(mu-H)(mu-BH(2)) 3-CH(2)CH(2)Ph-5-Ph-4,5-C(2)B(2)H(4), 6), (e) insertion and loss of borane with and without reduction (nido-1,2-(CpRu)(2)-5-Ph-4,5-C(2)B(2)H(7), 7, and isomers nido-1,2-(CpRu)(2)-3-CH(2)CH(2)Ph-4-(and-5-)Ph-C(2)B(2)H(6), 8 and 9), and (f) insertion and borane loss plus reduction (nido-1,2-(CpRu)(2)-3-(trans-CH=CHPh)-5 Ph-4,5-C(2)B(2)H(6), 10). Along with 7, 8, and 10, the reaction at 90 degrees C generates products of insertion and nido- to closo-cluster closure (closo-4-Ph 1,2-(CpRuH)(2)-4,6-C(2)B(2)H(3), 11, closo-1,2-(CpRuH)(2)-3-CH(2)CH(2)Ph-5-Ph-7 CH(2)CH(2)Ph-4,5-C(2)B(3)H(2), 12, closo-1,2-(CpRuH)(2)-5-Ph-4,5-C(2)B(3)H(4), 13, and isomers closo-1,2-(CpRuH)(2)-3-and-7-CH(2)CH(2)Ph-5-Ph-4,5-C(2)B(3)H(3), 14 and 15). The clusters with an exo-cluster bridging BH(2) groups are shown to be intermediates by demonstrating that the major products 5 and 6 rearrange to 13 and convert to 14, respectively. 14 then isomerizes to 15, thus connecting low- and high-temperature products. Finally, all available information shows that the high reactivity of 1 with alkynes can be associated with the "extra" two Ru-H hydrides on the framework of 1 which are required to meet the nido-cluster electron count. PMID- 15796549 TI - Syntheses and properties of two-dimensional charged nonlinear optical chromophores incorporating redox-switchable cis-tetraammineruthenium(II) centers. AB - In this article, we describe a series of new complex salts in which electron donating cis-{Ru(II)(NH3)(4)}(2+) centers are connected to two electron-accepting N-methyl/aryl-pyridinium groups. These V-shaped complexes contain either monodentate 4,4'-bipyridyl-derived ligands or related chelates based on 2,2':4,4":4',4'''-quaterpyridyl and have been characterized by using various techniques including electronic absorption spectroscopy and cyclic voltammetry. Molecular quadratic nonlinear optical (NLO) responses beta have been determined by using hyper-Rayleigh scattering at 800 nm and also via Stark (electroabsorption) spectroscopic studies on the intense, visible d --> pi metal to-ligand charge-transfer bands. These experiments reveal that these dipolar pseudo-C(2v)chromophores exhibit two substantial components of the beta tensor, beta(zzz)and beta(zyy), with the difference between them being most marked for the nonchelated systems. Time-dependent density-functional theory and finite field calculations serve to further illuminate the electronic structures and associated linear and NLO properties of the new chromophoric salts. PMID- 15796550 TI - Superexchange and sequential mechanisms in charge transfer with a mediating state between the donor and acceptor. AB - The rate of intramolecular charge transfer from biphenyl to naphthalene was determined for the radical anions and radical cations of molecules with the general structure: (2-naphthyl)-(steroid spacer)-(4-biphenylyl). Varied degrees of unsaturation (one double bond, NSenB; two double bonds, NSen(2)B; and the b ring completely aromatized, NSarB) were incorporated into the steroid spacer to examine the effect it would have on the charge transfer rate. The charge transfer rate, as inferred from the decay of the biphenyl radical ion absorption, increased in all cases relative to the completely saturated 3-(2-naphthyl)-16-(4 biphenylyl)-5alpha-androstane (NSB) reference molecule. For the anion charge transfer, the decay rates increased by factors of 1.4, 4.2, and 5.1, respectively, and for the cation, the decay rates increased by factors of 5, 276, and 470. To explain the results, the charge-transfer process was viewed as a combination of two independent mechanisms: a single-step, superexchange mechanism, and a two-step, sequential charge transfer. Using a low level of theory, simple models of the superexchange and two-step mechanisms were developed to elucidate the nature and differences between the two mechanisms. The critical variable for this analysis is the free energy of formation (DeltaG(I) degrees ) of the intermediate state: (2-naphthyl)-[spacer](1)+/--(4-biphenylyl). The conclusion from this treatment is that superexchange is the dominant mechanism when DeltaG(I) degrees is large, but at small DeltaG(I) degrees , the sequential mechanism will dominate. This is because the superexchange rate is shown to have a weak dependence on DeltaG(I) degrees , changing 10-fold for a change in DeltaG(I) degrees of 2 eV, compared to the sequential mechanism in which the rate can change over 10(3) for 0.5 V. PMID- 15796551 TI - Reactivity of titanium oxo ethoxo cluster [Ti16O16(OEt)32]. Versatile precursor of nanobuilding block-based hybrid materials. AB - Oxo alcoxo metallic clusters can be employed as inorganic nanobuilding blocks to obtain well-defined organic-inorganic hybrid materials. A better understanding of the surface reactivity of the clusters should allow optimization of the elaboration of hybrid materials through a better control of the hybrid interface. The oxo alcoxo cluster Ti(16)O(16)(OEt)32 presents a shell of labile ethoxy groups that can be selectively transalcoholyzed with preservation of the titanium oxo core, leading to new oxo alcoxo clusters Ti(16)O(16)(OEt)32-x(OR)x (R: alkyl, phenyl, styrenic, etc. groups). The reactivity of the Ti(16)O(16)(OEt)32 cluster toward aliphatic and aromatic alcohols is investigated to determine both the kinetics and the number of substituted titanium atoms, which are strongly dependent on the nature of the alcohol. Characterization of the organic modification of the cluster is performed in situ by liquid (13)C NMR measurements, using the molecular structures of two new clusters, Ti(16)O(16)(OEt)28(OnPr)4 and Ti(16)O(16)(OEt)(24)(OnPr)(8) (OnPr = propoxy groups), as references. The structures of these clusters have been established using single-crystal X-ray diffraction. Moreover, a complete spectroscopic assignment of each ethoxy group is proposed after combining crystallographic data, (13)C NMR T(1) relaxation measurements, and (1)H-(1)H, (1)H-(13)C 2D NMR experiments. Finally, the cluster is functionalized with polymerizable ligands via transalcoholysis and transesterification reactions using hydroxystyrene and acetoxystyrene. PMID- 15796552 TI - Structure of and competitive adsorption in alkyl dicarbamate two-dimensional crystals. AB - The potential for relatively minor structural changes to dramatically impact materials properties is one of the primary obstacles to achieving the rational design of functional materials. For example, having an odd versus an even number of carbons between functional groups in polymers can cause large variation in melting point and mechanical properties. This odd-even effect is especially pronounced in hydrogen-bonded polymers and oligomers. To shed light on the structural basis of this phenomenon, physisorbed monolayers and single crystals of alkyl dicarbamates were investigated by scanning tunneling microscopy and X ray diffraction, respectively. The related two- and three-dimensional crystal structures both demonstrated a clear odd-even effect in packing geometry. The differing accommodation of intermolecular interactions between odd and even packing motifs was directly related to the melting point trends and further dissected through computation. In addition, these oligomers displayed unusual competitive adsorption behavior; the relative preference for adsorption of a smaller species from a binary solution was increased compared to alkanes. These results were explained in the context of hydrogen bond density effects that arise due to competition for a limited substrate surface area. This study provides a model for understanding oligourethane surface coatings and demonstrates the importance of molecular structure and hydrogen bonding in determining adsorption behavior. PMID- 15796553 TI - Evolution of fluorescein as a platform for finely tunable fluorescence probes. AB - Fluorescence imaging is the most powerful technique currently available for continuous observation of dynamic intracellular processes in living cells. Suitable fluorescence probes are naturally of critical importance for fluorescence imaging, but only a very limited range of biomolecules can currently be visualized because of the lack of flexible design strategies for fluorescence probes. At present, design is largely empirical. Here we show that the carboxylic group of traditional fluorescein dyes, formerly considered indispensable, has been replaced with other substituents, affording various kinds of new fluoresceins. Further, by breaking out of the traditional structure of fluorescein, we developed the first and totally rational design strategy for novel fluorescence probes based on a strict photochemical basis. The value of this approach is exemplified by its application to develop a novel, highly sensitive, and membrane-permeable fluorescence probe for beta-galactosidase, which is the most widely used reporter enzyme. PMID- 15796554 TI - Modeling spin interactions in a cyclic trimer and a cuboidal Co4O4 core with Co(II) in tetrahedral and octahedral environments. AB - The X-ray crystallographic structures, the magnetic susceptibilities from 2 to 300 K, and a theoretical analysis of the magnetism for a triangular and a tetranuclear molecule consisting of linked high-spin cobalt(II) centers are described. The interpretation of the magnetic data for the triangular compound [Co(depa)Cl](3) (depa is the anion of 2,2'-(bis-4-ethylpyridyl)amine), which has tetrahedrally coordinated Co(2+) ions, entails isotropic antiferromagnetic exchange interaction and antisymmetric exchange acting within the two low-lying spin doublets. Two strong isotropic ferromagnetic interactions have been modeled in the cuboidal compound Co(4)(DPM)(4)(CH(3)O)(4)(CH(3)OH)(4) (DPM represents the anion of dipivaloylmethane), which has octahedral coordination, and the system can be approximately considered as two weakly coupled S = 3 species. PMID- 15796555 TI - Controlled electrophoretic patterning of polyaniline from a colloidal suspension. AB - We present a method for controlled deposition of polyaniline from colloidal suspensions. Stable suspensions of polyaniline colloids (approximately 115 nm in diameter) were formed by dispersing polyaniline/formic acid solution into acetonitrile. It was demonstrated that the positively charged polyaniline colloids can be electrophoretically deposited onto various substrate materials such as platinum and ITO, forming continuous ultrathin films. We examined the film morphology, as well as the effects of process parameters, such as deposition time, colloid concentration, and applied voltage, on the deposition efficiency. Furthermore, the efficacy of the technique was illustrated by electrophoretically patterning polyaniline thin films onto selected individual micrometer-scale sensing elements within a microfabricated sensor array, and by further demonstrating its sensitivity to gaseous analytes including water and methanol. PMID- 15796556 TI - Structure, recognition properties, and flexibility of the DNA.RNA hybrid. AB - Molecular dynamics is used to investigate the properties of the DNA.RNA hybrid in aqueous solution at room temperature. The structure of the hybrid is intermediate between A and B forms but, in general, closer to the canonical A-type helix. All the riboses exhibit North puckerings, while 2'-deoxyriboses exist in North, East, and South puckerings, the latter being the most populated one. The molecular recognition pattern of the DNA.RNA hybrid is a unique combination of those of normal DNA and RNA duplexes. Finally, the results obtained from essential dynamics and stiffness analysis demonstrate the large and very asymmetric flexibility of the hybrid and the strong predilection that each strand (DNA or RNA) has on the nature of their intrinsic motions in the corresponding homoduplexes. The implications of the unique structural and dynamic properties of the DNA.RNA hybrid on the mechanism of cleavage by RNase H are discussed. PMID- 15796557 TI - Zn protein simulations including charge transfer and local polarization effects. AB - Nearly half of all proteins contain metal ions, which perform a wide variety of specific functions associated with life processes. However, insights into the local/global, structural and dynamical fluctuations in metalloproteins from molecular dynamics simulations have been hampered by the "conventional" potential energy function (PEF) used in nonmetalloprotein simulations, which does not take into the nonnegligible charge transfer and polarization effects in many metal complexes. Here, we have carried out molecular dynamics simulations of Zn(2+) bound to Cys(-) and/or His(0) in proteins using both the conventional PEF and a novel PEF that accounts for the significant charge transfer and polarization effects in these Zn complexes. Simulations with the conventional PEF yield a nontetrahedral Cys(2)His(2) Zn-binding site and significantly overestimate the experimental Zn-S(Cys(-)) distance. In contrast, simulations with the new PEF accurately reproduce the experimentally observed tetrahedral structures of Cys(2)His(2) and Cys(4) Zn-binding sites in proteins, even when the simulation started from a nontetrahedral Zn(2+) configuration. This suggests that simulations with the new PEF could account for coordinational changes at Zn, which occurs during the folding/unfolding of Zn-finger proteins and certain enzymatic reactions The strategy introduced here can easily be applied to investigate Zn(2+) interacting with protein ligands other than Cys(-) and His(0). It can also be extended to study the interaction of other metals that have significant charge transfer and polarization effects. PMID- 15796558 TI - Electrocatalytic O2 reduction at glassy carbon electrodes modified with dendrimer encapsulated Pt nanoparticles. AB - Platinum dendrimer-encapsulated nanoparticles (DENs) were prepared within fourth generation, hydroxyl-terminated, poly(amidoamine) dendrimers and immobilized on glassy carbon electrodes using an electrochemical coupling strategy. X-ray photoelectron spectroscopy, electron microscopy, and electrochemical experiments confirmed that the Pt DENs were about 1.4 nm in diameter and that they remained within the dendrimer following surface immobilization. The resulting Pt DEN films were electrocatalytically active for the oxygen reduction reaction. The films were also robust, surviving up to 50 consecutive cyclic voltammograms and sonication. PMID- 15796559 TI - Use of molecular dynamics in the design and structure determination of a photoinducible beta-hairpin. AB - The study presented here consists of three parts. In the first, the ability of a set of differently substituted diazobenzene-based linkers to act as photoswitchable beta-turn building blocks was assessed. A 12-residue peptide known to form beta-hairpins was taken as the basis for the modeling process. The central (beta-turn) residue pair was successively replaced by six symmetrically ((o,o), (m,m), or (p,p)) substituted (aminomethyl/carboxymethyl or aminoethyl/carboxyethyl) diazobenzene derivatives leading to a set of peptides with a photoswitchable backbone conformation. The folding behavior of each peptide was then investigated by performing molecular dynamics simulations in water (4 ns) and in methanol (10 ns) at room temperature. The simulations suggest that (o,o)- and (m,m)-substituted linkers with a single methylene spacer are significantly better suited to act as photoswitchable beta-turn building blocks than the other linkers examined in this study. The peptide containing the (m,m) substituted linker was synthesized and characterized by NMR in its cis configuration. In the second part of this study, the structure of this peptide was refined using explicit-solvent simulations and NOE distance restraints, employing a variety of refinement protocols (instantaneous and time-averaged restraining as well as unrestrained simulations). We show that for this type of systems, even short simulations provide a significant improvement in our understanding of their structure if physically meaningful force fields are employed. In the third part, unrestrained explicit-solvent simulations starting from either the NMR model structure (75 ns) or a fully extended structure (25 ns) are shown to converge to a stable beta-hairpin. The resulting ensemble is in good agreement with experimental data, indicating successful structure prediction of the investigated hairpin by classical explicit-solvent molecular dynamics simulations. PMID- 15796560 TI - Variable-temperature X-ray crystallographic and DFT computational study of the N H...O/N...H-O tautomeric competition in 1-(Arylazo)-2-naphthols. Outline of a transition-state hydrogen-bond theory. AB - Phenyl-substituted 1-arylazo-2-naphthols (AAN) display ...HN-N=C-C=O... <==>...N=N-C=C-OH... ketohydrazone-azoenol tautomerism and can form intramolecular resonance-assisted H-bonds from pure N-H...O to pure N...H-O through tautomeric and dynamically disordered N-H...O <==>N...H-O bonds according to the electronic properties of their substituents. Three compounds of this series (m-OCH(3)-AAN = mOM; p-Cl-AAN = pCl; and p-NMe(2)-AAN = pNM2) have been studied by X-ray crystallography at four temperatures (100-295 K), showing that the remarkably short H-bonds formed (2.53 < or = d(N...O) < or = 2.55 A) are a pure N-H...O in mOM, a dynamically disordered mixture in pCl (N-H...O:N...H-O = 69:31 at 100 K), and a statically disordered mixture in pNM2 (N-H...O:N...H-O = 21:79 at 100 K). These compounds, integrated by the p-H-, p-NO(2)-, p-F-, and p O(-)-substituted derivatives, have been emulated by DFT methods (B3LYP/6 31+G(d,p) level) with full geometry optimization of the stationary points along the proton-transfer (PT) pathway: N-H...O and N...H-O ground states and N...H...O transition state. Analysis of DFT-calculated energies and geometries by the methods of the rate-equilibrium Marcus theory shows that all H-bond features (stability and tautomerism, as well as position and height of the PT barrier) can be coherently interpreted in the frame of the transition-state (or activated complex) theory by considering the bond as a chemical reaction N-H...O <==> N...H...O <==> N...H-O which is bimolecular in both directions and proceeds via the N...H...O PT transition state (the activated complex). PMID- 15796561 TI - CH5+: chemistry's chameleon unmasked. AB - The nuclear vibrational wave function and zero-point vibrational energy of CH5(+) are calculated using quantum diffusion Monte Carlo techniques on an interpolated potential energy surface constructed from CCSD(T)/aug'-cc-pVTZ ab initio data. From this multidimensional wave function, the vibrationally averaged rotational constants and radial distribution functions for atom-atom distances within the molecule are constructed. It is found that the distributions of all 10 H-H distances are bimodal and identical. The radial distribution functions obtained for the five C-H distances are also identical, but unimodal. The three rotational constants were found to be 3.78, 3.80, and 3.83 cm(-1). These values indicate that the ground state of CH5(+) is significantly more symmetric than its global minimum energy structure. We conclude that the zero-point motion of CH5(+) renders all five protons equivalent in the ground state and precludes the assignment of a unique structure to the molecule. PMID- 15796562 TI - Molecular dynamics simulation study on a monolayer of half [2]rotaxane self assembled on Au(111). AB - The self-assembled monolayer (SAM) structure of the tetrathiafulvalene-side half of the Stoddart-Heath type [2]rotaxane on Au(111) surface was investigated using molecular dynamics (MD) simulations. We find that the orientation of the cyclobis(paraquat-p-phenylene) (CBPQT) ring depends dramatically on the coverage, changing in order to obtain highly packed SAMs. The ring lies with its large hollow parallel to the surface at lower coverage (up to one CBPQT per 27 surface Au atoms with a footprint of 1.9 nm(2); 1/27) when free space is available around it, but as the coverage increases (up to one CBPQT per 12 surface Au atoms with a footprint of 0.9 nm(2); 1/12), it tilts completely around its axis and lies with its smaller side (paraquat or phenyl ring) parallel to the surface to accommodate the reduced area available. We find that the best packing densities correspond to one CBPQT per 12-18 surface Au atoms (1/18-1/12) with footprints in the range between 0.9 nm(2) and 1.3 nm(2). PMID- 15796564 TI - Ion gels prepared by in situ radical polymerization of vinyl monomers in an ionic liquid and their characterization as polymer electrolytes. AB - To realize polymer electrolytes with high ionic conductivity, we exploited the high ionic conductivity of an ionic liquid. In situ free radical polymerization of compatible vinyl monomers in a room temperature ionic liquid, 1-ethyl-3-methyl imidazolium bis(trifluoromethane sulfonyl)imide (EMITFSI), afforded a novel series of polymer electrolytes. Polymer gels obtained by the polymerization of methyl methacrylate (MMA) in EMITFSI in the presence of a small amount of a cross linker gave self-standing, flexible, and transparent films. The glass transition temperatures of the gels, which we named "ion gels", decreased with increasing mole fraction of EMITFSI and behaved as a completely compatible binary system of poly(methyl methacrylate) (PMMA) and EMITFSI. The temperature dependence of the ionic conductivity of the ion gels followed the Vogel-Tamman-Fulcher (VTF) equation, and the ionic conductivity at ambient temperature reached a value close to 10(-2) S cm(-1). Similarly to the behavior of the ionic liquid, the cation in the ion gels diffused faster than the anion. The number of carrier ions, calculated from the Nernst-Einstein equation, was found to increase for an ion gel from the corresponding value for the ionic liquid itself. The cation transference number increased with decreasing EMITFSI concentration due to interaction between the PMMA matrix and the TFSI(-) anion, which prohibited the formation of ion clusters or associates, as was the case for the ionic liquid itself. PMID- 15796563 TI - On the mechanism of THF catalyzed vinylic lithiation of allylamine derivatives: structural studies using 2-D and diffusion-ordered NMR spectroscopy. AB - N-Lithio-N-(trialkylsilyl)allylamines can be deprotonated in the presence of ethereal solvents exclusively at the cis-vinylic position to yield 3,N-dilithio-N (trialkylsilyl)allylamines under mild conditions. Low temperature (1)H and (7)Li NMR ((1)H NOESY, TOCSY, (1)H/(7)Li HSQC, and DO-NMR) studies on the solution structure of 3,N-dilithio-N-(tert-butyldimethylsilyl)allylamine identified three major aggregates in THF (monomer, dimer and tetramer), but the aggregate structures failed to explain the solvent dependence and regiochemical outcome of the reaction. Low temperature (1)H NMR (NOESY, TOCSY, DO-NMR) studies on the solution structure of N-lithio-N-(tert-butyldimethylsilyl)allylamine in the presence of nBuLi identified amide/nBuLi mixed aggregates in both the ethereal solvent THF (1:1 dimer) and the hydrocarbon solvent toluene (1:3 tetramer). Addition of 2 equiv of THF to toluene solutions induces the formation of the same THF solvated 1:1 dimer as observed in neat THF. NMR evidence suggests that in THF the mixed aggregate has close contact between the olefin and the beta-CH(2) of nBuLi, while in the absence of THF, the allyl chain appears to be pointed away from the nearest nBuLi residues. PMID- 15796565 TI - Estimation of color of durum wheat. Comparison of WSB, HPLC, and reflectance colorimeter measurements. AB - Color is an important parameter involved in the definition of semolina and pasta quality. This character is mainly due to natural pigments (carotenoids) that are present at different levels in cereals and cereal products, due to botanical origin, growing conditions, distribution in the kernel, and technological processes. In food industries, color measurements are usually performed by means of automatic instruments that are rapid and safe, as alternatives to the chemical extraction methods. In this study, automatic measurements (CIE, color-space system L, a, b), water-saturated butanol (WSB), and HPLC determinations have been applied to evaluate the carotenoid content in whole meals and respective semolina samples produced from wheat cultivated in the years 2001 and 2002. In whole meals, total carotenoids, determined by HPLC, were about 3.0 microg/g (2001) and 3.5 microg/g (2002) calculated on dry weight (dw) and about 3.0 and 3.2 microg/g dw in corresponding semolina samples. The b values for the same period were 19.78 and 15.75, respectively, in raw materials and 20.03-21.67 in semolina. Results have confirmed lutein and beta-carotene as the main components mainly responsible for the yellow color in wheat grains. The ability of the index b to express natural dyeing was dependent on sample characteristics as demonstrated by the relationships found between this index and pigments, although the best correlation resulted between HPLC and WSB. PMID- 15796567 TI - Analysis of lidocaine and its major metabolite, monoethylglycinexylidide, in elk velvet antler by liquid chromatography with UV detection and confirmation by electrospray ionization tandem mass spectrometry. AB - A sensitive liquid chromatographic (LC) method with UV detection was developed for the determination of residues of lidocaine (LID) and its major metabolite, monoethylglycinexylidide (MEGX), in elk velvet antler. The drugs were extracted from alkaline velvet antler homogenates, cleaned up on a C(18) solid-phase extraction cartridge, and separated on an Inertsil ODS-3 (3.0 x 250 mm, 5 microm) column using an isocratic mobile phase made up of 0.05 M phosphate buffer (pH 4.0)/acetonitrile (88:12, v/v) at a flow rate of 1.0 mL/min. The limits of quantification for LID and its major metabolite, MEGX, were 10 and 20 ng/g, respectively. The method was validated and used to measure the concentration of residues of LID and MEGX in elk velvet antlers harvested after either LID anesthesia or application of a drug-free control method (electro-anesthesia, EA). No LID or MEGX residues were detected in any of the antlers harvested after EA application. No MEGX residues were detected in any of the velvet antlers harvested after LID application, but residues of LID ranging in concentration from 68 to 4300 ng/g were detected in the three sections of the velvet antlers harvested after LID administration. LC-tandem mass spectrometry was used to confirm the presence of lidocaine detected in the velvet antlers. PMID- 15796566 TI - Development of an ELISA for quantifying lysozyme in hen egg white. AB - An indirect enzyme-linked immunosorbent assay (ELISA) by inhibition was developed for quantifying lysozyme in hen egg white (HEW), a protein of value in not only the food and pharmaceutical industries but also for poultry research. Various experimental conditions (coating, antibodies dilutions, samples dilutions, preparations, blocking agents, and incubation times) were assayed to optimize this assay to the quantification of HEW in egg white samples. HEW samples were diluted 1:3000 to avoid matrix effects, possibly resulting from lysozyme interaction with other egg white proteins. Assay linearity for lysozyme ranged from 0.38 to 4.8 mug/mL, with intra- and interassay variations of 6.8% and 7.6%, respectively, and the lower detection limit was 0.264 mug/mL. We found that lysozyme concentrations in albumen from eggs laid by a hen cohort ranged from 2.2 to 4.5 mg/mL, thus underlining interhen variability. Overall, these data present an ELISA assay that is simple, quick, sensitive, accurate, and has been specifically designed to determine lysozyme concentrations in egg white samples. PMID- 15796568 TI - Application of bubble separation for quantitative analysis of choline in Dioscorea (yam) tubers. AB - A modified assay based on the AACC official method 86-45 (AACC, 2000) for the determination of choline in three cereals and three varieties of Dioscorea (yam) tubers was developed. When tested in wheat, rice, and oat flour, choline estimated by the modified method was 34.0-45.3% higher than that of the original AACC method. In a system with higher contents of starch and mucilage, such as Dioscorea (yam) tubers, extra procedures in sample preparation needed to be carried out to separate starch and mucilage. The choline contents of the following Dioscorea (yam) tubers using the original AACC method and the present modified AACC method through coupling an additional bubble separation procedure, respectively, were (mean +/- SD, mg/g solid) Keelung yam (D. pseudojaponicaY.) 0.92 +/- 0.09 and 2.21 +/- 0.12, Yangmingshan yam (D. alata L.) 0.77 +/- 0.09 and 1.78 +/- 0.28, and Ming-Chien yam (D. purpurea) 0.44 +/- 0.09 and 1.35 +/- 0.19. Choline was 231-306% higher than when the original AACC method was used. Dioscorea (yam) tubers were much higher in choline content than they were in cereals. Bubble separation is an appropriate procedure in the practice for the maximum assay of choline in yams. It is accurate, rapid, easy to handle, and especially good for recovering choline from a starch and polysaccharide-protein containing system. PMID- 15796569 TI - Evaluation of pheromone release from commercial mating disruption dispensers. AB - Pome fruit growers and crop consultants have expressed concerns about the seasonal release performance of commercial codling moth mating disruption dispenser products. Because of these concerns, we developed a laboratory flow through volatile collection system (VCS) for measuring the volatile release of the codling moth sex pheromone, codlemone, from commercially available hand applied dispensers. Under controlled air-flow and temperature conditions, the released vapor was trapped onto a polyurethane foam adsorbent followed by solvent extraction, solvent reduction, and GC/MS determination. Method recovery and breakthrough validations were performed to demonstrate system reliability before determining codlemone release from commercial dispensers field-aged over 140 days. The volatile collection was carried out in a consistent manner among five dispenser types most commonly used by growers, so that direct comparison of performance could be made. The comparison showed differences in the amount of pheromone released and in the patterns of release throughout the season between dispenser types. The variation in release performance demonstrates the need for routine evaluation of commercially marketed mating disruption dispensers. We believe that the simple and cost-effective volatile collection system can assist pheromone dispenser manufacturers in determining seasonal dispenser performance before new products are introduced into the commercial market and in rapidly verifying dispenser release when field-aged dispenser efficacy is in question. PMID- 15796570 TI - Development of a simplified method for the determination of folates in baker's yeast by HPLC with ultraviolet and fluorescence detection. AB - A simplified HPLC method for rapid determination of folates in yeast with ultraviolet and fluorescence detection without sample purification has been developed. By use of the column Aquasil C(18), specially designed for polar analytes, and gradient elution, it was possible to separate and determine five folate derivatives: tetrahydrofolate, 5-methyltetrahydrofolate, and 5 formyltetrahydrofolate with fluorescence detection, and 10-formylfolic acid and folic acid with ultraviolet detection. The sample preparation required only a small amount of dry yeast (25-50 mg) and included an extraction of folates by heat treatment and deconjugation of folate polyglutamates to monoglutamates with the use of rat serum conjugase. Validation involved investigation of matrix effects, determination of recovery by standard addition method, repeatability, and stability tests. The dominating folate forms in commercial dry baker's yeast were found to be tetrahydrafolate and 5-methyltetrahydrofolate with a total folate content of 2890 microg/100 g (63.4 nmol/g). The simplicity of the method makes it suitable for folate screening studies of different yeast strains. PMID- 15796571 TI - Determination of the age of sherry wines by regression techniques using routine parameters and phenolic and volatile compounds. AB - This paper describes a study of the possibility of obtaining regression models by means of partial least squares (PLS) and multiple linear regression (MLR) that would enable us to correlate a series of parameters, such as the concentration of short-chain organic acids, higher alcohols, and phenolic compounds with the age of vintage Sherry wines or "anadas". The aim of this study is to characterize how these parameters evolve with aging. If this could be done, it would then be possible to guarantee the age of such wines using objective variables. A PLS regression model was established that allows the age to be predicted with a mean deviation of 1.6 years with respect to the age of the wines. In the case of the MLR, a model with 6 variables was obtained that gives a mean deviation of 3.3 years in the predictions. PMID- 15796572 TI - Quantitation of dityrosine in wheat flour and dough by liquid chromatography tandem mass spectrometry. AB - A method for the quantitation of dityrosine in wheat flour and dough by high performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) using an isotope dilution assay with the internal standard 3,3'-(13)C(2)-dityrosine in the single-reaction monitoring mode was developed. The method consisted of the release of protein-bound dityrosine by hydrolysis in 4 mol/L hydrochloric acid/8.9 mol/L propionic acid for 24 h at 110 degrees C after addition of the internal standard, cleanup by C(18) solid-phase extraction, and HPLC-MS/MS. The limit of detection of dityrosine was 80 ng/g of sample (0.22 nmol/g), and the limit of quantitation was 270 ng/g of sample (0.75 nmol/g). The method was sensitive enough to analyze wheat flour and dough and to study the effect of flour improvers on the dityrosine content. Furthermore, the effect of the mixing time was studied. The dityrosine concentration in the flour was 0.66 nmol/g. After we mixed a dough to peak consistency, the dityrosine concentration doubled and remained constant on further mixing. Overdoses of hydrogen peroxide and hexose oxidase (HOX, E.C. 1.1.3.5) resulted in a strongly increased dityrosine content, whereas no increase of the dityrosine concentration was found after the addition of ascorbic acid and potassium bromate. Calculation of the percentage of dimeric tyrosine showed that less than 0.1% of the tyrosine residues of wheat protein were cross-linked. Therefore, dityrosine residues seem to play only a very minor role in the structure of wheat gluten. PMID- 15796573 TI - Authentication and quantitative analysis on the chemical profile of cassia bark (cortex cinnamomi) by high-pressure liquid chromatography. AB - Cassia bark or cortex cinnamomi, the dried stem bark of Cinnamomum cassia Presl. (Lauraceae), is a popular natural spice and a commonly used herb in traditional Chinese medicine. However, adulterants are frequently found in the market. In this study, 44 samples of Cassia bark including bark from seven related Cinnamomum species were collected from fields and market. Four characteristic components, cinnamaldehyde, cinnamic acid, cinnamyl alcohol, and coumarin were determined by RP-HPLC, and a fingerprint comprised of five markers was established. These results showed that cassia barks contained high contents of cinnamaldehyde (13.01-56.93 mg/g). The highest content of cinnamaldehyde (up to 93.83 mg/g) was found in debarked cortex, which is traditionally regarded as having the best quality in local herb shops. In contrast, the adulterants from the other Cinnamomum species, C. wilsonii Camble, C. japonicum Sieb., C. mairei Levl. and C. burmanii (Nees) Blume, contained low contents of cinnamaldehyde (<2.00 mg/g). The content of cinnamaldehyde in C. loureirii Nees was comparable to that in C. cassia. It is suggested that five characteristic peaks by HPLC are suitable for distinguishing genuine cassia bark from the adulterants and could be applied in the quality control of this commodity. PMID- 15796574 TI - Screening of Yucatecan plant extracts to control Colletotrichum gloeosporioides and isolation of a New Pimarene from Acacia pennatula. AB - Screening of seven Yucatecan plant extracts to look for fungicidal activity for the control of Colletotrichum gloeosporioides was carried out. Bioassay-directed purification of the root extract of one of the most active plants, Acacia pennatula, resulted in the isolation of the new compound 15,16-dihydroxypimar 8(14)-en-3-one (1), which in the in vitro bioassay "agar dilution" was shown to have growth, sporulation, and germination inhibition activity. Nuclear magnetic resonance spectroscopic techniques were used to elucidate its structure. PMID- 15796575 TI - Antioxidant activity of commercial soft and hard wheat (Triticum aestivum L.) as affected by gastric pH conditions. AB - Phenolic compounds from soft and hard wheat and their milling fractions were extracted into distilled deionized water, and their in vitro antioxidant activities were evaluated. Wheat samples were used as such (nontreated) or subjected to pH adjustment (treated) in order to simulate gastrointestinal pH conditions. The total phenolic content (TPC) was determined using Folin Ciocalteu's procedure. The total antioxidant activity (TAA) was determined using Trolox equivalent antioxidant capacity assay and expressed as Trolox equivalents. The antioxidant activity of wheat extracts was also evaluated using the beta carotene bleaching assay, scavenging of 2,2-diphenyl-1-picrylhydrazyl radical, and inhibition of oxidation of human low density lipoprotein cholesterol. The TPC, TAA, and antioxidant potential, evaluated using different methods of wheat samples, were significantly increased following gastrointestinal tract-simulated pH changes. Thus, digestion taking place in the gastrointestinal tract in vivo may also enhance the antioxidant properties of the extracts. PMID- 15796576 TI - Isolation, characterization, and determination of antioxidative activity of oligomeric procyanidins from the seedpod of Nelumbo nucifera Gaertn. AB - The procyanidins of nonedible parts of lotus (Nelumbo nucifera Gaertn.) were determined for the first time. The procyanidins of lotus seedpod were extracted with Me(2)CO/H(2)O and purified by Sephadex LH-20 column chromatography, with a purity of >98%. ESI-MS analysis showed that the main molecular weight distribution of procyanidins ranged from 291 to 1155, with M + H peak values of 291.1, 579.2, 731.2, 867.2, 1019.4, and 1155.3, respectively. This indicates that the extract contains monomers, dimers, and tetramers of procyanidins, in which the amounts of dimers are greatest, and catechin and epicatechin are the base units. (1)H NMR and (13)C NMR spectra confirmed that two to four monomers are linked through C(4)-C(8) (or C(6)) bonds. The effects of the procyanidins on lipid autoxidation, lipoxygenase activities, and free radical scavenging were also studied. The results showed that 0.1% procyanidins have a strong antioxidant activity in a soybean oil system, better than BHT at the same concentration; inhibited lipoxygenase activity by >90% at a concentration of 62.5 mug/mL, with an IC(50) value of 21.6 mug/mL; and had IC(50) inhibitory values rate to (*)OH of 10.5 mg/L and a scavenging effect on O(2)(*)(-) of 17.6 mg/L. PMID- 15796577 TI - Cuminaldehyde: Aldose Reductase and alpha-Glucosidase Inhibitor Derived from Cuminum cyminum L. Seeds. AB - The inhibitory activity of Cuminum cyminum seed-isolated component was evaluated against lens aldose reductase and alpha-glucosidase isolated from Sprague-Dawley male rats and compared to that of 11 commercially available components derived from C. cyminum seed oil, as well as quercitrin as an aldose reductase inhibitor and acarbose as an alpha-glucosidase inhibitor. The biologically active constituent of C. cyminum seed oil was characterized as cuminaldehyde by various spectral analyses. The IC(50) value of cuminaldehyde is 0.00085 mg/mL against aldose reductase and 0.5 mg/mL against alpha-glucosidase, respectively. Cuminaldehyde was about 1.8 and 1.6 times less in inhibitory activity than acarbose and quercitin, respectively. Nonetheless, cuminaldehyde may be useful as a lead compound and a new agent for antidiabetic therapeutics. PMID- 15796578 TI - Enantioselective synthesis of the female sex pheromone of the pink hibiscus Mealybug, Maconellicoccus hirsutus. AB - The pink hibiscus mealybug, Maconellicoccus hirsutus (Green), is an exotic insect pest and recently invaded Southern California and Florida. The female M. hirsutus releases the 2-methylbutanoate of a novel cyclobutanoid monoterpene alcohol (maconelliol) that together with lavandulyl 2-methylbutanoate constitutes the sex pheromone to attract males from a distance. Enantioselective syntheses of four different stereoisomers of the major component, maconelliyl 2-methylbutanoate 1, from alpha-pinene are reported. Absolute configurations of both naturally occurring maconelliyl 2-methylbutanoate 1 and the minor component, lavandulyl 2 methylbutanoate 2, have been established. Comparison of the analytical data of naturally occurring compounds with those of optically active synthetic isomers proved that both esters show the (R)-configuration at the alcohol and the (S) configuration at acid moieties. PMID- 15796579 TI - Evidence for formation of heterooligosaccharides by Lactobacillus sanfranciscensis during growth in wheat sourdough. AB - Lactobacillus sanfranciscensis is a key organism of the lactic microflora in traditional and industrial sourdough fermentations. In this paper we provide evidence for the formation of heterooligosaccharides (HeOS) by L. sanfranciscensis during growth in sourdough. To identify the HeOS based on HPAEC PAD analysis, HeOS standards were synthesized by enzymatic reactions with L. sanfranciscensis levansucrase in a chemically defined system in the presence of raffinose, maltotriose, maltose, xylose, or arabinose as acceptor carbohydrates. The oligosaccharides known to originate from the corresponding acceptor reactions, 1(F)-beta-fructosylraffinose, 1(F)-beta-fructofuranosylmaltotriose, erlose (1(F)-beta-fructofuranosylmaltose), xylsucrose, 1(F)-beta fructosylxylsucrose, and arabsucrose, were identified by HPAEC-PAD. Evidence for the formation of further tri-, tetra-, and pentasaccharides was provided. Wheat doughs with sucrose were fermented with L. sanfranciscesis TMW 1.392 or the isogenic, levansucrase-negative strain TMW 1.392Deltalev, and the analysis of dough extracts or invertase-treated dough extracts provided evidence for the formation of arabsucrose and erlose in sourdough in addition to 1-kestose and nystose. PMID- 15796580 TI - Hypocholesterolemic effects of a flavonoid-rich extract of Hypericum perforatum L. in rats fed a cholesterol-rich diet. AB - In a previous study, a flavonoid-rich extract of Hypericum perforatum L. (FEHP) was prepared and its antioxidant activity was determined by a series of models in vitro. In this study, the hypocholesterolemic effects of FEHP in rats fed a cholesterol-rich diet were tested. Forty Wistar rats fed a standard laboratory diet or a cholesterol-rich diet for 16 weeks were used. The serum lipid levels, as well as malondialdehyde (MDA) and activity of superoxide dismutase (SOD) and catalase (CAT) in serum and liver, were examined. Cholesterol-rich diet induced hypercholesterolemia was manifested in the elevation of serum lipid levels such as total cholesterol (TC), total triglycerides (TG), and low density lipoprotein cholesterol (LDL-C). Administration of middle-dose (75 mg/kg of BW/day) and high dose (150 mg/kg of BW/day) FEHP significantly lowered the serum levels of TC, TG, and LDL-C, while increasing the serum level of high density lipoprotein cholesterol (HDL-C). Also, the content of MDA in serum and liver decreased significantly after oral administration of FEHP compared with those of rats fed a cholesterol-rich diet. In addition, FEHP increased the activity of SOD in serum and liver, but the activity of CAT was significantly elevated only in liver. These results suggested that the hypocholesterolemic effects of FEHP might be due to its abilities to lower serum TC, TG, and LDL-C levels as well as to slow the lipid peroxidation process and to enhance the antioxidant enzyme activity. PMID- 15796581 TI - Antimutagenic and antioxidant properties of milk-kefir and soymilk-kefir. AB - This study was aimed at evaluating the antimutagenic and antioxidant properties of milk-kefir and soymilk-kefir. Such antimutagenic activity was determined by means of the Salmonella mutagenicity assay, whereas the antioxidant properties of kefir were evaluated by assessing the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity, lipid peroxidation inhibition activity, ferrous ion chelating ability, reducing power, and antioxidative enzyme activity. Both milk kefir and soymilk-kefir demonstrated significantly greater antimutagenic activity than milk and soymilk. Milk-kefir and soymilk-kefir also displayed significantly greater scavenging activity upon DPPH radicals, an inhibition effect upon linoleic acid peroxidation, and more substantial reducing power but displayed a reduced glutathione peroxidase activity than was the case for milk and soymilk. Milk and soymilk fermented by kefir grains did not alter the ferrous ion chelating ability and superoxide dismutase activity of the original materials. These findings have demonstrated that milk-kefir and soymilk-kefir possess significant antimutagenic and antioxidant activity and suggest that milk-kefir and soymilk-kefir may be considered among the more promising food components in terms of preventing mutagenic and oxidative damage. PMID- 15796582 TI - Direct antioxidant activity of purified glucoerucin, the dietary secondary metabolite contained in rocket (Eruca sativa Mill.) seeds and sprouts. AB - Rocket (Eruca sativa Mill. or Eruca vesicaria L.) is widely distributed all over the world and is usually consumed fresh (leafs or sprouts) for its typical spicy taste. Nevertheless, it is mentioned in traditional pharmacopoeia and ancient literature for several therapeutic properties, and it does contain a number of health promoting agents including carotenoids, vitamin C, fibers, flavonoids, and glucosinolates (GLs). The latter phytochemicals have recently gained attention as being the precursors of isothiocyanates (ITCs), which are released by myrosinase hydrolysis during cutting, chewing, or processing of the vegetable. ITCs are recognized as potent inducers of phase II enzymes (e.g., glutathione transferases, NAD(P)H:quinone reductase, epoxide hydrolase, etc.), which are important in the detoxification of electrophiles and protection against oxidative stress. The major GL found in rocket seeds is glucoerucin, GER (108 +/- 5 micromol g(-)(1) d.w.) that represents 95% of total GLs. The content is largely conserved in sprouts (79% of total GLs), and GER is still present to some extent in adult leaves. Unlike other GLs (e.g., glucoraphanin, the bio-precursor of sulforaphane), GER possesses good direct as well as indirect antioxidant activity. GER (and its metabolite erucin, ERN) effectively decomposes hydrogen peroxide and alkyl hydroperoxides with second-order rate constants of k(2) = 6.9 +/- 0.1 x 10(-)(2) M(-)(1) s(-)(1) and 4.5 +/- 0.2 x 10(-)(3) M(-)(1) s(-) , respectively, in water at 37 degrees C, thereby acting as a peroxide-scavenging preventive antioxidant. Interestingly, upon removal of H(2)O(2) or hydroperoxides, ERN is converted into sulforaphane, the most effective inducer of phase II enzymes among ITCs. On the other hand, ERN (and conceivably GER), like other ITCs, does not possess any chain-breaking antioxidant activity, being unable to protect styrene from its thermally (37 degrees C) initiated autoxidation in the presence of AMVN. The mechanism and relevance of the antioxidant activity of GER and ERN are discussed. PMID- 15796583 TI - Benoxacor induction of terbuthylazine detoxification in Zea mays and Festuca arundinacea. AB - The action of safener benoxacor on the detoxification of terbuthylazine (TBA) in Zea mays and Festuca arundinacea was ascertained by the investigation of the effects of benoxacor on the activity of glutathione-S-transferases (GSTs) in the shoots of the two plant species. TBA treatment generally reduced GST activity toward 1-chloro-2,4-dinitrobenzene (CDNB) in corn and did not affect the enzyme activity in festuca. When applied alone, benoxacor stimulated GST activity in both plants; however, when it was applied in mixture with TBA, generally, an enhancement of the enzyme activity was found in corn but not in festuca in comparison with the respective TBA-treated samples. The enhancement of GST activity in response to the benoxacor treatment in both corn and festuca resulted to be concomitant with decreases in apparent K(M) in both the plant species, with V(max) unaffected, and with an increased expression of proteins having molecular masses in the characteristic range of plant GSTs. After the benoxacor treatment, increased GST activity toward TBA as a substrate was observed in both corn and festuca. As a consequence, lesser amounts and persistence of TBA residues were found in shoots of corn and festuca treated with the TBA and benoxacor mixture compared to TBA-only-treated samples. Therefore, benoxacor enhances TBA detoxification in both corn and festuca; the induction of detoxifying activity in a nondomesticated grass is discussed in view of its use in vegetating buffer strips around crops to prevent TBA pollution. PMID- 15796585 TI - Food-drug interaction between ferulic acid and nateglinide involving the fluorescein/H+ cotransport system. AB - In clinical, patients usually take many kinds of drugs at the same time. Thus, drug-drug interactions involving transporters can often directly affect the therapeutic safety and efficacy of many drugs. However, there have been few studies on food-drug interactions involving transporters. Dietary polyphenols have been widely assumed to be beneficial to human health. Polyphenols are commercially prepared and used as functional foods. We report here for the first time that ferulic acid, which is widely used as a functional food, affects the transport of clinical agents. It is important to be aware of the potential of food-drug interactions and to act in order to prevent undesirable and harmful clinical consequences. PMID- 15796584 TI - Effects of grape cell culture extracts on human topoisomerase II catalytic activity and characterization of active fractions. AB - Grape and its cell culture extracts are rich in flavonoids and stilbenes that are biologically active. The objective of this study was to evaluate possible inhibitory effects of grape (a Vitis hybrid Bailey Alicant A) cell culture extract and subfractions on human DNA topoisomerase II catalytic activity and to characterize constituents in the most potent fractions. At 5 microg/mL, grape cell crude extract and Toyopearl (TP) fractions 2-6 provided significantly greater inhibition of topoisomerase II catalytic activity than quercetin, a chemopreventive agent previously known as a topoisomerase catalytic inhibitor. The most potent topoisomerase II catalytic inhibitors from grape cell culture extracts in descending order of potency were TP fractions 4 and 6 (IC(50) = 0.28 0.29 microg/mL), TP-3 (IC(50) = 0.74 microg/mL), and crude extract (IC(50) = 1.02 microg/mL); each was significantly more potent than resveratrol (IC(50) = 18.0 microg/mL), another well-known chemopreventive topoisomerase II catalytic inhibitor. Using both high-performance liquid chromatography and liquid chromatography electrospray ionization mass spectrometry, constituents in TP-4 and TP-6 were characterized. These constituents included cyanidin-3,5 diglucoside, malvidin-3-acetylglucoside, peonidin-3-coumaryl-5-diglucoside, procyanidin B(1), procyanidin B(2), procyanidin B(5), procyanidin dimer digallate, procyanidin C(1), myricetin, and rutin, none of which have been previously characterized from grape cell cultures. The significant potency especially of TP-4 and TP-6 from grape cell cultures suggests that these fractions may have potential as chemopreventive agents. PMID- 15796586 TI - Antioxidant activity of steryl ferulate extracts from rye and wheat bran. AB - Antioxidant activity of steryl ferulates from other sources than rice have not yet been studied much, despite the fact that rice steryl ferulates (gamma oryzanol) have been shown to possess good antioxidant activity. In this study, steryl ferulate extracts from wheat or rye bran were studied for their capability to inhibit hydroperoxide formation in bulk methyl linoleate and methyl linoleate emulsion. Further, their activity to scavenge DPPH radicals was analyzed. The activities were compared to synthetic steryl ferulates, rice steryl ferulates, ferulic acid, and alpha-tocopherol. Nonrice cereal extracts of steryl ferulates exhibited good antioxidant activity, especially in the bulk lipid system. The radical scavenging activity was similar to that of nonesterified ferulic acid, indicating that the ferulic acid moiety is responsible for the antioxidant properties. This study illustrates a new aspect to the health-promoting properties of rye and wheat. PMID- 15796587 TI - Free radical scavengers and antioxidants from Lemongrass (Cymbopogon citratus (DC.) Stapf.). AB - Methanol, MeOH/water extracts, infusion, and decoction of Cymbopogon citratus were assessed for free radical scavenging effects measured by the bleaching of the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical, scavenging of the superoxide anion, and inhibition of the enzyme xanthine oxidase (XO) and lipid peroxidation in human erythrocytes. The extracts presented effect in the DPPH and superoxide anion assay, with values ranging between 40 and 68% and 15-32% at 33 and 50 microg/mL, respectively, inhibited lipid peroxidation in erythrocytes by 19-71% at 500 microg/mL and were inactive toward the XO at 50 microg/mL. Isoorientin, isoscoparin, swertiajaponin, isoorientin 2' '-O-rhamnoside, orientin, chlorogenic acid, and caffeic acid were isolated and identified by spectroscopic methods. Isoorientin and orientin presented similar activities toward the DPPH (IC(50): 9 10 microM) and inhibited lipid peroxidation by 70% at 100 microg/mL. Caffeic and chlorogenic acid were active superoxide anion scavengers with IC(50) values of 68.8 and 54.2 microM, respectively, and a strong effect toward DPPH. Caffeic acid inhibited lipid peroxidation by 85% at 100 microg/mL. PMID- 15796588 TI - Polyacetylenes from the Apiaceae vegetables carrot, celery, fennel, parsley, and parsnip and their cytotoxic activities. AB - A dichloromethane extract of root celery yielded falcarinol, falcarindiol, panaxydiol, and the new polyacetylene 8-O-methylfalcarindiol. The structure of the new compound was established by one- and two-dimensional (1D and 2D) NMR, mass spectrometry, and optical rotation data. Nonpolar extracts of roots and bulbs of carrots, celery, fennel, parsley, and parsnip were investigated for their content of polyacetylenes by high-performance liquid chromatography with diode array detection (HPLC-DAD). All five species contained polyacetylenes, although carrots and fennel only in minor amounts. Additionally, the cytotoxicity of the four polyacetylenes against five different cell lines was evaluated by the annexin V-PI assay. Falcarinol proved to be the most active compound with a pronounced toxicity against acute lymphoblastic leukemia cell line CEM-C7H2, with an IC(50) of 3.5 micromol/L. The possible chemopreventive impact of the presented findings is discussed briefly. PMID- 15796589 TI - Bioautography and chemical characterization of antimicrobial compound(s) in commercial water-soluble annatto extracts. AB - Annatto preparations based on extracts of the seed of tropical bush Bixa orellana L consist of carotenoid-type pigments. Previous reports indicate that commercial annatto extracts have biological activities against microorganisms of significance to food fermentation, preservation, and safety. The objective of this study was to separate and identify the compound(s) responsible for the antimicrobial activity of annatto preparations. Commercial water-soluble annatto extracts were screened by thin-layer chromatography and bioautography followed by liquid chromatography/photodiode array/mass spectrometry (LC/PDA/MS) analysis of active fractions. Bioautography revealed two fractions with antimicrobial activity against Staphylococcus aureus. LC/PDA/MS analysis of both fractions revealed 9'-cis-norbixin (UV(max) 460 and 489 nm) and all-trans-norbixin (UV(max) 287, 470, and 494 nm) as the major components. Structure confirmation was achieved by (1)H NMR spectroscopy. Results indicate that 9'-cis-norbixin and all trans-norbixin are responsible for the antimicrobial properties of annatto. PMID- 15796590 TI - Modulation of cytokine secretion by garlic oil derivatives is associated with suppressed nitric oxide production in stimulated macrophages. AB - We previously described that garlic oil derivatives differentially suppress the production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in activated macrophages. In the present study, we investigated the effects of the garlic derivatives, diallyl sulfide (DAS), diallyl disulfide (DADS), and allyl methyl sulfide (AMS), on cytokine production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and the association between modulation of cytokines and inhibition of NO production was also assessed. The results indicated that these garlic compounds had different effects on the secretion of activated cytokines, including proinflammatory tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, and IL-6, as well as the antiinflammatory, IL-10. DAS inhibited the production of all stimulated cytokines in a concentration-dependent manner, and the inhibition was closely associated with the suppression of NO and PGE(2) production. DADS repressed the production of stimulated TNF-alpha and IL-10 and increased the production of activated IL-1beta and, to a lesser extent, IL-6; but only the decreased IL-10 production was associated with DADS-induced NO inhibition. Yet, the DAS- and DADS-suppressed NO production was independent of TNF-alpha. AMS, on the other hand, slightly suppressed the stimulated TNF-alpha but enhanced IL-10 production, and such modulation was closely associated with the decrease in NO production. PMID- 15796591 TI - Quantification of genetically modified soybean by quenching probe polymerase chain reaction. AB - Quenching probe (QProbe) polymerase chain reaction (PCR) is a simple and cost effective real-time PCR assay in comparison with other real-time PCR assays such as the TaqMan assay. We used QProbe-PCR to quantify genetically modified (GM) soybean (Roundup Ready soybean). We designed event-specific QProbes for Le1 (soy endogenous gene) and RRS (recombinant gene), and we quantified certified reference materials containing 0.1, 0.5, 1, 2, and 5% GM soybean. The TaqMan assay was also applied to the same samples, and the results were compared. The accuracy of QProbe-PCR was similar to that of TaqMan assay. When GM soybean content was 0.5% or more, the relative standard deviations of QProbe-PCR were less than 20%. QProbe-PCR is sensitive enough to monitor labeling systems and has acceptable levels of accuracy and precision. PMID- 15796592 TI - Enhancement of natural pigment extraction using Bacillus species xylanase. AB - Pigment extracts from the root of Lithospermum erythrorhizon are used as natural red dyes, as well as basic drugs due to their numerous pharmacological activities. In recent years, the demand for such natural pigment materials has increased; however, in natural dye production, the pigment yield is strongly affected by the source of cultivation, extracting conditions, and solvents. Accordingly, this study proposes a method of enzymatic pigment production based on the introduction of hydrolytic enzymes prior to the usual extraction to avoid repeated pigment extraction. The matrix destruction in the epidermal layer of the root by the enzymes was found to improve the pigment extractability, that is, the increment of K(L), the mass transfer coefficient, representing the pigment mobility in the epidermal layer. The root tissue maceration by the hydrolytic enzymes was also measured to evaluate the pigment extractability, and a linear relationship was observed between the K(L) values and the tissue maceration up to the addition of 3000 units/g of xylanase, indicating that the enzymatic maceration proportionally increases the interfacial area between the pigment and the solvent. Bacillus sp. DX107 xylanase only served to increase the extractability of the pigment by loosening the root shell matrix, without affecting the contents and color properties of the pigment, as almost no difference was found in the color between the pigments extracted using xylanase and those extracted according to the traditional method. PMID- 15796593 TI - Malolactic fermentation in wine with Lactobacillus casei cells immobilized on Delignified cellulosic material. AB - In this work Lactobacillus casei ATCC 393 cells immobilized on delignified cellulosic material (DCM) were used for malolactic fermentation (MLF) of wine. Wine was produced using yeast cells immobilized on DCM at 20 degrees C, and after alcoholic fermentation, MLF at 27 degrees C followed using immobilized L. casei ATCC 393 cells. A total of 11 repeated alcoholic and subsequent MLF batches were performed within a period of 1 month. As the repeated MLF batches proceeded, the MLF activity of the immobilized biocatalyst was reduced. Malic acid degradation was reduced from 80 to 2%, pH was reduced by 0.5-0.1 unit, acetic acid concentrations were slightly reduced or remained stable (0.002 g/L), the higher alcohols 1-propanol, isobutyl alcohol, and amyl alcohol were decreased by 84, 23, and 11%, respectively, and ethyl acetate concentration was increased by approximately 56%. Wine samples were analyzed by GC-MS before and after MLF, revealing some qualitative differences. PMID- 15796594 TI - Biochemical origin of browning during the processing of fresh Yam (Dioscorea spp.) into dried product. AB - This study was undertaken to follow the kinetics of polyphenoloxidase (PPO), peroxidase (POD), and phenolic compounds during yam blanching at different temperatures and after drying and to identify by high-performance liquid chromatography (HPLC) the main phenolic compounds present in yam products. PPO activity was 50% higher in nonprocessed freeze-dried Florido (Dioscorea alata) than in nonprocessed freeze-dried Deba (Dioscorea rotundata). It decreased progressively during blanching. Forty-five percent of PPO activity remained after 50 min of blanching at 60 or 65 degrees C, whereas the POD activity dropped sharply to less than 20% of the initial activity after 10 min of blanching, whatever the blanching temperature. No anthocyanidins could be detected by HPLC at 520 nm in nonprocessed freeze-dried yam. Flavanols with a maximum absorption wavelength (lambda(max)) at 280 nm and cinnamic acid compounds with 320 nm lambda(max) were detected. Catechin was identified as the major flavanol with concentrations ranging from 0.26 to 0.41 microM g(-)(1) depending on cultivar. One cinnamic compound, ferulic acid, was identified and assessed in both cultivars (0.03-0.04 microM g(-)(1)). Total phenol, flavanol, and cinnamic contents decreased during blanching independently of temperature whereas some unresolved peaks were detected by HPLC in dried product. The latter was probably due to the consumption of coloring precursors and the appearance of polymerized or complex colored products. PMID- 15796595 TI - Ultrastructural changes and structure and mobility of myowater in frozen-stored hake (Merluccius merluccius L.) muscle: relationship with functionality and texture. AB - The ultrastructural changes and the main Raman spectral features of water (3100 3500 and 50-600 cm(-)(1) ranges) in frozen-stored hake were studied with the aim of connecting these changes with loss of some functional properties such as water holding capacity, and with modifications of muscle texture. The following results were obtained: (a) The changes in the spaces between myofibrils can be related to modifications of shear resistance. (b) The behavior of the strong 160 cm(-)(1) band can be related to conformational transitions of muscle proteins, to changes in the structure of muscle water, and/or to alterations in protein-water interactions. (c) There were intensity changes in the nu(s)(OH) band that may be attributable to transfer of water to larger spatial domains during frozen storage. PMID- 15796596 TI - Oil quality and sensory evaluation of almond (Prunus amygdalus) stored after electron beam processing. AB - The changes in the lipid fraction and the deterioration of its quality were studied in almonds (Prunus amygdalus) of the variety Guara after treatment with accelerated electrons at doses of 3, 7, and 10 kGy, during a storage period of 5 months. In almond oil, the most significant difference from the nutritional point of view was seen in the fatty acid linolenic (18:3), which shows at 3 kGy a maintenance of the initial content during the whole storage period, whereas, at 7 and 10 kGy, the content in 18:3 disappears from the first moment. The quality indices of the oil (K(232), K(270)) decreased at all doses and remained stable during the time of storage. The peroxide value did not show changes at the doses of 3 and 7 kGy, in non-irradiated samples, but significantly increased when the maximum dose of 10 kGy was applied. These changes were reflected in the sensory analysis, in which the tasters did not find sensory differences between the controls and those irradiated at doses of 3 or 7 kGy, whereas almonds irradiated at 10 kGy exhibited a rancid flavor and a significant decrease in general quality. PMID- 15796597 TI - Multivariate correlation between color and mineral composition of honeys and by their botanical origin. AB - The mineral content and color characteristics of 77 honey samples were analyzed. Eighteen minerals were quantified for each honey. Multiple linear regression (MLR) was used to establish equations relating the colorimetric CIELAB coordinates to the mineral data. The results obtained shown that lightness (L) was significantly correlated with S, Ca, Fe, As, Pb, and Cd for the dark honey types (avocado, heather, chestnut, and honeydew). For the light and brown honey types (citrus, rosemary, lavender, eucalyptus, and thyme), C(ab) and b showed the lower correlation with the mineral content of the honeys; their regression functions involve a few independent variables (Mg and Al for b and only Al for C(ab)). Furthermore, by means of application of linear discriminant analysis to the mineral content, it was possible to obtain a model that classifies the honeys by their lightness. The prediction ability of the built model, determined with the test set method, was 85%. PMID- 15796598 TI - Evaluation of the effect of germination on phenolic compounds and antioxidant activities in sorghum varieties. AB - The screening of 50 sorghum varieties showed that, on average, germination did not affect the content in total phenolic compounds but decreased the content of proanthocyanidins, 3-deoxyanthocyanidins, and flavan-4-ols. Independent of germination, there are intervarietal differences in antioxidant activities among sorghum varieties. Phenolic compounds and antioxidant activities were more positively correlated in ungerminated varieties than in germinated ones. Sorghum grains with pigmented testa layer, chestnut color glumes, and red plants had higher contents, larger diversity of phenolic compounds, and higher antioxidant activities than other sorghums. Some red sorghum varieties had higher antioxidant activities (30-80 mumol of Trolox equiv/g) than several sources of natural antioxidants from plant foods. Among varieties used for "to", "dolo", couscous, and porridge preparation, the "dolo"(local beer) varieties had the highest average content and diversity in phenolic compounds as well as the highest antioxidant activities. The biochemical markers determined are useful indicators for the selection of sorghum varieties for food and agronomic properties. PMID- 15796599 TI - Systematic identification and characterization of anthocyanins by HPLC-ESI-MS/MS in common foods in the United States: fruits and berries. AB - Anthocyanins were systematically identified and characterized by HPLC-ESI-MS/MS coupled with diode array detection in common fruits from U.S. food markets and other commercial sources. Of the 25 different fruits that were screened, 14 fruits were found to contain anthocyanins; the number of anthocyanins varied from 2 in peaches and nectarines to 31 in Concord grape. The individual anthocyanins were identified by comparing their mass spectral data and retention times with those of standards and published data. In all of the samples analyzed, only 6 common anthocyanidins, delphinidin, cyanidin, pelargonidin, petunidin, peonidin and malvidin, were found. In addition to the well-known major anthocyanins, a number of minor anthocyanins were identified for the first time. Some possible guidelines that help to identify anthocyanins in foods with complex anthocyanin composition were deduced and discussed. For the first time, this paper presents complete anthocyanin HPLC profiles and MS spectral data of common fruits using the same uniform experimental conditions. PMID- 15796600 TI - Fibrillar microfilaments associated with a high-amylose rice, goami 2, a mutant of ilpumbyeo, a high-quality japonica rice. AB - The ultrastructure of cooked and malt-treated cooked rice of Ilpumbyeo (IP) and its mutant Goami 2 (G2), which have extreme contrasts in physicochemical properties, cooking quality, and ultrastructural characteristics in raw grains (1, 2), was compared. In cooked rice of IP, starch granules in endosperm cells were evenly coalesced, appearing as homogeneously smooth sheetlike matrix and/or globules, whereas those in G2 were a heterogeneously coarse matrix in which a novel structural feature, the microfilaments, was embedded. In malt-treated cooked rice of IP, most starch was hydrolyzed by the malt enzymes, appearing as empty vacuoles surrounded by the cell wall, whereas that in G2 was highly resistant to malt treatment, remaining as distinct structural features, the malt resistant compound starch granules. The property of G2's compound starch granules, which are tolerant of mechanical and chemical treatments thereby retaining their structural integrity (2) and of cooking and malt treatment thereby retaining their physical hardness, appears to play a major role in determining the quality of cooked rice of G2. PMID- 15796602 TI - Comparison of volatiles of cultured and wild sea bream (Sparus aurata) during storage in ice by dynamic headspace analysis/gas chromatography-mass spectrometry. AB - Cultured and wild sea bream were compared for differences in their volatile components over a 23 day storage period in ice. A total of 60 compounds in cultured and 78 compounds in wild sea bream were tentatively identified (in addition to this, there were 23 unknowns in cultured and 29 unknowns in wild sea bream volatiles). These included aldehydes, ketones, alcohols, aromatics, terpenes, furans, sulfur-containing compounds, an acid, and miscellaneous compounds. Although selection of best fish is a subjective matter, more aldehydes, ketones, aromatics, and terpenes were found in wild sea bream as compared to that of its cultured counterpart. Both sea bream samples exhibited complex volatile profiles over the entire storage period. The combination of several classes of volatile compounds, dependent upon their concentrations and odor thresholds, is responsible for the distinctive and unique flavor of fresh cultured and wild sea bream. Relative concentrations of several compounds (trimethylamine, piperidine, methanethiol, dimethyl disulfide, dimethyl trisulfide, 1-penten-3-ol, 3-methyl-1-butanol, and acetic acid) increased continually throughout the storage period, and these may have the potential to be used as indicators of sea bream quality. PMID- 15796601 TI - Accelerators increase permeability of cuticles for the lipophilic solutes metribuzin and iprovalicarb but not for hydrophilic methyl glucose. AB - Effects of diethylsuberate (DESU), tributyl phosphate (TBP), and monodisperse ethoxylated alcohols (EAs) on rate constants of penetration (k) of model solutes across astomatous cuticular membranes isolated from Madagascar ivy (Stephanotis floribunda) and pear (Pyrus communis) leaves were studied. Model solutes (selected on the basis of their octanol/water partition coefficients, K(ow)) were iprovalicarb (log K(ow) = 3.18), metribuzin (log K(ow) = 1.60), and methyl glucose (MG) (log K(ow) = -3.0). K(ow) varied by more than 6 orders of magnitude. Accelerators had wax/water partition coefficients (log K(ww)) ranging from 1.75 (DESU) to 4.32 (C(12)E(2)), and their equilibrium concentrations in Stephanotis wax varied from 0 to about 160 g kg(-)(1). Accelerators increase solute mobility in cuticles by increasing fluidity of cutin and waxes. This effect was quantified by plotting log k versus the accelerator concentration in wax. With the lipophilic solutes metribuzin and iprovalicarb, these plots were linear. Slopes of these plots characterize the intrinsic activities of the accelerators, and they decreased in the order DESU (0.029) > TBP (0.015) > EAs (0.01). Using these intrinsic activities, the effects of accelerators on rate constants of penetration can be calculated for any accelerator concentration in wax. For instance, at 50 g kg(-)(1), rate constants for lipophilic solutes increased by factors of 28 (DESU), 5.6 (TBP), and 3.2 (C(12)E(n)()), respectively. Permeability of cuticles for the hydrophilic MG was not increased by DESU, TBP, C(12)E(2), and C(12)E(4), while C(12)E(6) and C(12)E(8) increased it. Small hydrophilic solutes such as MG can access aqueous pores in cuticles, and this pathway is not affected by changes in fluidity of amorphous waxes. After rate constants of penetration of ionic CaCl(2) were compared with those for nonionic MG, it was concluded that 60% of the MG diffused across aqueous pores, while 40% used an alternative pathway. Because the solubility of MG in wax is extremely low, it is unlikely that MG diffused along the lipophilic pathway used by metribuzin and iprovalicarb. This agrees with the observation that DESU and TBP had no effect on rate constants for MG. An alternative pathway of unknown properties is suggested. It is speculated that C(12)E(6) and C(12)E(8) sorbed in cuticles might have generated a polar pathway for MG. PMID- 15796603 TI - Screening of raw coffee for thiol binding site precursors using "in bean" model roasting experiments. AB - The purpose of the following study was to investigate the influence of coffee roasting on the thiol-binding activity of coffee beverages, and to investigate the potential of various green bean compounds as precursors of thiol-binding sites by using promising "in bean" model roast experiments. Headspace gas chromatographic analysis on coffee brews incubated in the presence of the roasty sulfury smelling 2-furfurylthiol for 20 min at 30 degrees C in septum-closed vessels revealed that the amounts of "free" thiol decreased drastically with increasing the roasting degree of the beans used for preparation of the brews. A half-maximal binding capacity (BC(50)) of 183 mg of 2-furfurylthiol per liter of standard coffee beverage was determined for a roasted coffee (CTN value of 67), thus demonstrating that enormous amounts of the odor-active thiol are "bound" by the coffee. Furthermore, biomimetic "in bean" precursor experiments have been performed in order to elucidate the precursor for the thiol-binding sites in the raw coffee bean. These experiments opened the possibility of studying coffee model reactions under quasi-natural roasting conditions and undoubtedly identified chlorogenic acids as well as thermal degradation products caffeic acid and quinic acid as important precursors for low-molecular-weight thiol-binding sites. In particular, when roasted in the presence of transition metal ions, chlorogenic acids and even more caffeic acid showed thiol-binding activity which was comparable to the activity measured for the authentic coffee brew. PMID- 15796604 TI - Chemical composition of Juniperus communis L. fruits supercritical CO2 extracts: dependence on pressure and extraction time. AB - Ground fruits of the common juniper (Juniperus communis L.), with a particle size range from 0.250-0.400 mm, forming a bed of around 20.00 +/- 0.05 g, were extracted with supercritical CO(2) at pressures of 80, 90, and 100 bars and at a temperature of 40 degrees C. The total amount of extractable substances or global yield (mass of extract/mass of raw material) for the supercritical fluid extraction process varied from 0.65 to 4.00% (wt). At each investigated pressure, supercritical CO(2) extract fractions collected in successive time intervals over the course of the extraction were analyzed by capillary gas chromatography, using flame ionization (GC-FID) and mass spectrometric detection (GC-MS). More than 200 constituents were detected in the extracts, and the contents of 50 compounds were reported in the work. Dependence of the percentage yields of monoterpene, sesquiterpene, oxygenated monoterpene, and oxygenated sesquiterpene hydrocarbon groups on the extraction time was investigated, and conditions that favored the yielding of each terpene groups were emphasized. At all pressures, monoterpene hydrocarbons were almost completely extracted from the berries in the first 0.6 h. It was possible to extract oxygenated monoterpenes at 100 bar in 0.5 h and at 90 bar in 1.2 h. Contrary to that, during an extraction period of 4 h at 80 bar, it was possible to extract only 75% of the maximum yielded value of oxygenated monoterpene at 100 bar. Intensive extraction of sesquiterpenes could be by no means avoided at any pressure, but at the beginning of the process (the first 0.5 h) at 80 bar, they were extracted about 8 and 3 times slower than at 100 and 90 bar, respectively. Oxygenated sesquiterpenes were yielded at fast, constant extraction rates at 100 and 90 bar in 1.2 and 3 h, respectively. This initial fast extraction period was consequently followed by much slower extraction of oxygenated sesquiterpenes. PMID- 15796605 TI - Ability of possible DMS precursors to release DMS during wine aging and in the conditions of heat-alkaline treatment. AB - The origin of dimethyl sulfide (DMS) produced during wine aging was examined through different assays. The production of DMS during the model aging of a wine and the concomitant decrease of residual potential DMS (PDMS), as DMS released by heat-alkaline treatment in 0.5 M sodium hydroxide at 100 degrees C for 1 h, were demonstrated. Then, dimethyl sulfoxide (DMSO), methionine sulfoxide (MSO), S methylmethionine (SMM), and dimethylsulfonium propanoic acid (DMSPA), reported previously as possible DMS precursors, were investigated for their ability to be DMS precursors in wine in the conditions of this model aging and of the heat alkaline treatment. The results showed that DMSO, MSO, and DMSPA could hardly be DMS precursors in the conditions used, whereas SMM appeared to be a good candidate. Finally, the use of [(2)H(6)]-DMSPA as an internal standard for PDMS determination was proposed, because it provided better reproducibility than [(2)H(6)]-DMS used as an external standard. PMID- 15796606 TI - Partitioning behavior of Alkan-1-ols between milkfat and aqueous phases as influenced by temperature. AB - Partitioning of volatile compounds between lipid and aqueous phases may influence flavor perception and availability to participate in flavor-generating reactions. The objective of this research was to characterize the partitioning of short chain alkan-1-ols between milkfat and aqueous phases, as influenced by temperature, as compared to an octan-1-ol/water biphasic system (Log P). Temperature has a positive, but nonlinear, influence on Log P values for alkan-1 ols. There is an approximately 1 log decrease in Log P values of milkfat/water as compared to octan-1-ol/water systems; similar trends were observed across chain length. Temperature has a greater effect on alkan-1-ol partitioning in milkfat/water systems than octan-1-ol/water. The latter observation is primarily attributed to the solidification of milkfat at temperatures below 40 degrees C and the resulting reduction in liquid lipid solvent volume. PMID- 15796607 TI - Induction of de novo volatile terpene biosynthesis via cytosolic and plastidial pathways by methyl jasmonate in foliage of Vitis vinifera L. AB - The terpene biosynthesis in leaves of Vitis vinifera L. cv. Morio Muskat was studied using methyl jasmonate to induce defensive responses in vivo. The experiments demonstrated the strong activation of the de novo biosynthesis of terpenoids via the octadecanoid-signaling cascade and release of the compounds to the gas phase. Feeding experiments with [5,5-(2)H(2)]-1-deoxy-d-xylulose and [5,5 (2)H(2)]mevalonic acid lactone allowed the investigation of the dynamic allocation of resources via the mevalonic acid and 1-deoxy-d-xylulose/2-C-methyl d-erythritol 4-phosphate (DOXP/MEP) pathway under induced conditions and after treatment with the specific inhibitors mevastatin and fosmidomycin. The experiments reveal that monoterpenes are almost exclusively synthesized via the DOXP/MEP pathway, whereas sesquiterpenes are generated via both pathways at approximately equal rates. The biosynthesis of the homoterpene (E)-4,8-dimethyl 1,3,7-nonatriene was not affected by mevastatine or fosmidomycin. PMID- 15796608 TI - Antioxidant properties of roasted coffee residues. AB - The antioxidant activity of roasted coffee residues was evaluated. Extraction with four solvents (water, methanol, ethanol, and n-hexane) showed that water extracts of roasted coffee residues (WERCR) produced higher yields and gave better protection for lipid peroxidation. WERCR showed a remarkable protective effect on oxidative damage of protein. In addition, WERCR showed scavenging of free radicals as well as the reducing ability and to bind ferrous ions, indicating that WERCR acts as both primary and secondary antioxidants. The HPLC analyses showed that phenolic acids (chlorogenic acid and caffeic acid) and nonphenolic compounds [caffeine, trigonelline, nicotinic acid, and 5 (hydroxymethyl)furfuraldehyde] remained in roasted coffee residues. These compounds showed a protective effect on a liposome model system. The concentrations of flavonoids and polyphenolic compounds in roasted coffee residues were 8,400 and 20,400 ppm, respectively. In addition, the Maillard reaction products (MRPs) remaining in roasted coffee residues were believed to show antioxidant activity. These data indicate that roasted coffee residues have excellent potential for use as a natural antioxidant source because the antioxidant compounds remained in roasted coffee residues. PMID- 15796609 TI - Nitrogen metabolism components as a tool to discriminate between organic and conventional citrus fruits. AB - The aim of this work was to develop a method for authenticity control of organically grown orange fruits. Due to the different kinds of nitrogen fertilization of the soil in organically and conventionally managed farms, the study tried to verify the possibility to differentiate Navelina and Tarocco orange fruits obtained by these production systems through the detection of markers linked to nitrogen metabolism. In addition to the classic quality parameters, total nitrogen (N) and synephrine contents in juice and (15)N/(14)N isotope ratio (expressed as delta(15)N per thousand) in proteins of pulp and amino acids of juice were determined. The results obtained indicated that total N and synephrine contents were significantly higher in conventional fruits, whereas the delta(15)N per thousand values were higher in the organic ones. The new markers identified in this research by linear discriminant analysis of the data may constitute a useful tool to differentiate organic citrus fruits or juices from conventional ones. PMID- 15796610 TI - Casein proteins as molecular chaperones. AB - Under conditions of stress, such as elevated temperature, molecular chaperones stabilize proteins from unfolding, aggregating, and precipitating. We have investigated the chaperone activity of the major milk proteins alpha(S)-, beta-, and kappa-casein with reduced insulin and the milk whey proteins, alpha lactalbumin and beta-lactoglobulin, and compared it with that of the mammalian small heat shock protein (sHsp), alpha-crystallin, and clusterin. alpha(S)-Casein exhibited different chaperone behavior under reduction and heat stresses, i.e., chaperone activity increased with increasing temperature (as observed with alpha crystallin), but under reduction stress, its chaperone activity increased at lower temperatures. beta- and kappa-casein had comparable chaperone ability with each other but were less effective than alpha(S)-casein. Under molecular crowding conditions, precipitation of stressed protein was accelerated, and alpha(S) casein was a poorer chaperone. Furthermore, at slightly alkaline pH values, alpha(S)-casein was a less effective chaperone than at neutral pH. Detailed fluorescence, size exclusion chromatography, and real-time NMR studies studies indicated that the casein proteins underwent conformational changes and stabilized the partially unfolded whey proteins prior to formation of high molecular weight soluble complexes. These results are consistent with casein proteins acting as molecular chaperones in a manner similar to sHsps and clusterin. PMID- 15796611 TI - Effect of roasting on properties of the zinc-chelating substance in coffee brews. AB - ApV is a brownish polymer with zinc-chelating activity in brewed coffee. We investigated in this study the effects of roasting on the zinc-chelating, reducing, and antioxidative activities of ApV from light-, medium-, and dark roasted coffee. We also discuss the effect on the zinc-chelating activity of adding milk to the brewed coffee. The chelating activities of ApVs were evaluated by the tetramethyl murexide method. As the intensity of roasting increased, the yield of ApV increased, and the brown color and molecular weight of ApV respectively became darker and higher. Increasing the degree of roasting also decreased the zinc-chelating activity of ApV. The reducing activities of ApVs estimated by the indophenol method were stronger than those of ascorbic acid. Both the antioxidative activity estimated by the ABTS assay and the reducing activity of ApV increased with roasting. When milk was added to instant coffee and its ApV was prepared, the zinc-chelating activity of ApV was not changed. PMID- 15796612 TI - Conjugated linoleic acid in processed cheeses during the manufacturing stages. AB - Conjugated linoleic acid (CLA) is a naturally occurring micronutrient in milk fat and dairy products consisting of a group of geometric and positional isomers. The purpose of this study was to assess the level and type of CLA isomers found in two commercial processed cheeses (portions and slices) as well as to monitor their evolution during the different manufacturing stages. Total CLA concentrations ranged from 7.5 to 7.9 mg/g of fat, and rumenic acid (cis-9,trans 11 C18:2), the isomer responsible for the biological functions, represented >80% of total CLA. trans-11,cis-13 and trans-11,trans-13 were, with approximately 4% each, the second main CLA isomers. trans-trans isomers accounted for <10% of total CLA. The processing parameters used in this research had negligible effects on the CLA content of processed cheese and did not modify the isomer profile in these dairy products, thereby confirming the stability of rumenic acid during manufacturing. PMID- 15796613 TI - Free radical reactions and antioxidant activities of sesamol: pulse radiolytic and biochemical studies. AB - Sesamol (from Sesamum indicum) is a dietary compound, which is soluble in aqueous as well as lipid phases. Free radical scavenging reactions of sesamol, 5-hydroxy 1,3-benzodioxole, have been studied using a nanosecond pulse radiolysis technique. Sesamol efficiently scavenges hydroxyl, one-electron oxidizing, organo haloperoxyl, lipid peroxyl, and tryptophanyl radicals. Its antioxidant activity has also been evaluated with cyclic voltammetry. In biochemical studies, it has been found to inhibit lipid peroxidation, hydroxyl radical-induced deoxyribose degradation, and DNA cleavage. These antioxidant and free radical scavenging activities of sesamol have been reported in the paper. PMID- 15796614 TI - Interactions of ferric ions with olive oil phenolic compounds. AB - The ferric complexing capacity of four phenolic compounds, occurring in olives and virgin olive oil, namely, oleuropein, hydroxytyrosol, 3,4 dihydroxyphenylethanol-elenolic acid (3,4-DHPEA-EA), and 3,4 dihydroxyphenylethanol-elenolic acid dialdehyde (3,4-DHPEA-EDA), and their stability in the presence of ferric ions were studied. At pH 3.5, all compounds formed a reversible 1:1 complex with ferric ions, but hydroxytyrosol could also form complexes containing >1 ferric ion per phenol molecule. At pH 5.5, the complexes between ferric ions and 3,4-DHPEA-EA or 3,4-DHPEA-EDA were relatively stable, indicating that the antioxidant activity of 3,4-DHPEA-EA or 3,4-DHPEA-EDA at pH 5.5 is partly due to their metal-chelating activity. At pH 7.4, a complex containing >1 ferric ion per phenol molecule was formed with hydroxytyrosol. Oleuropein, 3,4-DHPEA-EA, and 3,4-DHPEA-EDA also formed insoluble complexes at this pH. There was no evidence for chelation of Fe(II) by hydroxytyrosol or its derivatives. At all pH values tested, hydroxytyrosol was the most stable compound in the absence of Fe(III) but the most sensitive to the presence of Fe(III). PMID- 15796615 TI - Very long chain fatty acid synthesis in sunflower kernels. AB - Most common seed oils contain small amounts of very long chain fatty acids (VLCFAs), the main components of oils from species such as Brassica napus or Lunnaria annua. These fatty acids are synthesized from acyl-CoA precursors in the endoplasmic reticulum through the activity of a dissociated enzyme complex known as fatty acid elongase. We studied the synthesis of the arachidic, behenic, and lignoceric VLCFAs in sunflower kernels, in which they account for 1-3% of the saturated fatty acids. These VLCFAs are synthesized from 18:0-CoA by membrane bound fatty acid elongases, and their biosynthesis is mainly dependent on NADPH equivalents. Two condensing enzymes appear to be responsible for the synthesis of VLCFAs in sunflower kernels, beta-ketoacyl-CoA synthase-I (KCS-I) and beta ketoacyl-CoA synthase-II (KCS-II). Both of these enzymes were resolved by ion exchange chromatography and display different substrate specificities. While KCS I displays a preference for 20:0-CoA, 18:0-CoA was more efficiently elongated by KCS-II. Both enzymes have different sensitivities to pH and Triton X-100, and their kinetic properties indicate that both are strongly inhibited by the presence of their substrates. In light of these results, the VLCFA composition of sunflower oil is considered in relation to that in other commercially exploited oils. PMID- 15796617 TI - Comparison of the bioactive compounds and antioxidant potentials of fresh and cooked Polish, Ukrainian, and Israeli garlic. AB - Garlic (Allium sativum L.) is an essential part of Polish, Ukrainian, and Israeli cuisine. The aim of this investigation was to compare the changes in bioactive compounds, proteins, and antioxidant potentials in fresh Polish, Ukrainian, and Israeli garlic samples after subjection to cooking temperature. Dietary fiber and essential trace elements were comparable. The antioxidant potentials were determined by four scavenging methods using beta-carotene, 1,1-diphenyl-2 picrylhydrazyl (DPPH), nitric oxide (NO), and 2,2'-azinobis(3 ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS(*)(+)) radical cation with K(2)S(2)O(8) or MnO(2) assays. Polyphenols, tocopherols, proteins, and antioxidant potentials were higher in Polish garlic, but not significantly (P > 0.05). The SDS- and native-PAGE electrophoretic patterns of all three fresh garlic samples were without significant differences. Most of the proteins were in the molecular mass range of 24-97 kDa, and the more intensive major bands were concentrated at 50 and 12 kDa. The 50 kDa protein nearly disappears and the intensity of the 12 kDa lectin bands slightly decreases during cooking. It was observed that the bioactive compounds, antioxidant potential, and proteins in garlic decrease significantly after 20 min of cooking at 100 degrees C (P < 0.05). In conclusion, (a) the bioactive compounds, electrophoretic patterns, and antioxidant potential of fresh Polish, Ukrainian, and Israeli garlic samples are comparable; (b) garlic samples subjected to 100 degrees C during 20 min preserve their bioactive compounds, antioxidant potential, and protein profile and are comparable with fresh garlic; and (c) fresh garlic should be added to dishes cooked at 100 degrees C in the last 20 min of the cooking process. PMID- 15796618 TI - Flavonol glucoside profile of southern Italian red onion (Allium cepa L.). AB - High-performance liquid chromatography-diode array detector (HPLC-DAD) coupled with electron spray mass spectrometry (ESI-MS-MS) was used to determine the flavonol profile in southern Italian red onions (Allium cepa L.). This on-line technique allowed the identification of seven flavonols in southern Italian red onion, quercetin 4'-glucoside and quercetin 3,4'-diglucoside being the most abundant components. Five minor flavonols have been recognized, offering a characteristic profile of such compounds in red onions under study. Quercetin 3 glucoside, quercetin 7,4'-diglucoside, quercetin 3,7,4'-triglucoside, and isorhamnetin 4'-glucoside have been previously reported as minor flavonoid components in Allium cepa, while isorhamnetin 3,4'-diglucoside was previously found in Allium ascalonicum. Traces of isorhamnetin 3-glucoside and free quercetin were also detected. PMID- 15796616 TI - Influence of amylopectin structure and amylose content on the gelling properties of five cultivars of cassava starches. AB - Five cassava genotypes were investigated to identify the fine amylopectin structures and granule chemical compositions, which differentiated the starches into high (T(o) = 63.7 degrees C on average) and low (57.3 degrees C on average) gelatinization temperatures. The amylose contents (15.9-22.4%) and granular dimensions (12.9-17.2 microm) significantly differed among the starches. Diverse amylopectin structural elements resulted in significant swelling power, viscoelastic properties, and gel firmness. Debranched starches revealed a trimodal amylopectin distribution of three fractions: FIII (DP 12), FII (DP 24.31), and FI (DP 63) and FIII (DP 12), FII (DP 24.69), and FI (DP 67) for the low and high gelatinization starch groups, respectively. The higher proportion of FI long chain entanglement with amylose chain lengths to form longer helical structures was confirmed in the high gelatinization starch group, which developed "true" gels with better shear resistance, frequency independence, and higher gel firmness. Significant amounts of resistant starch fractions revealed the potential for application of these genotype starches in diverse foods. PMID- 15796619 TI - Chemical constituents and antioxidant activity of sweet cherry at different ripening stages. AB - The development and ripening process of sweet cherry (Prunus avium L. cv. 4-70) on the tree was evaluated. For this purpose, 14 different stages were selected in accordance with homogeneous size and color. Some parameters related to fruit quality, such as color, texture, sugars, organic acids, total antioxidant activity, total phenolic compounds, anthocyanins, and ascorbic acid were analyzed. The results revealed that in sweet cherry, the changes in skin color, glucose and fructose accumulation, and softening process are initiated at early developmental stages, coinciding with the fast increase in fruit size. Also, the decrease in color parameter a was correlated with the greatest accumulation of total anthocyanins. Ascorbic acid, total antioxidant activity (TAA), and total phenolic compounds decreased during the early stages of sweet cherry development but exponentially increased from stage 8, which coincided with the anthocyanin accumulation and fruit darkening. TAA showed positive correlations (r(2) = 0.99) with both ascorbic acid and total phenolic compounds and also with the anthocyanin concentration from stage 8. Taking into account the reduced shelf life of sweet cherry and to ensure that these fruits reach consumers with the maximum organoleptic, nutritional, and functional properties, it is advisable to harvest sweet cherries at stage 12 of ripening. PMID- 15796620 TI - Effects of chlorophyll-related compounds on hydrogen peroxide induced DNA damage within human lymphocytes. AB - Chlorophylls (Chl's) are the most abundant natural plant pigments. Four chlorophyll-related compounds (CRCs), including chlorophyllide a and b (Chlide a and b) and pheophorbide a and b (Pho a and b), were investigated for their antioxidative capacities to protect human lymphocyte DNA from hydrogen peroxide (H(2)O(2)) induced strand breaks and oxidative damage ex vivo. Lymphocytes exposed to H(2)O(2) at concentrations of 10 and 50 microM revealed an increased frequency of DNA single-strand breaks (ssb's; as measured by the comet assay) and also an increased level of oxidized nucleoside (as measured by 8 hydroxydeoxyguanosine, 8-OHdG). All Chl's reduced the level of DNA ssb's and 8 OHdG within human lymphocytes following exposure to 10 microM H(2)O(2). Only Pho a and b were able to decrease DNA ssb's and 8-OHdG following treatment of lymphocytes with 50 microM H(2)O(2), in a concentration-dependent fashion. It was demonstrated herein that Pho a and b were more antioxidative than others. We applied DPPH free-radical scavenge assays in vitro, and got similar results. Pho a and b had higher ability in scavenging capacities than others. We conclude that water-extract Chl's are able to enhance the ability of human lymphocytes to resist H(2)O(2)-induced oxidative damage, especially for Pho a and b. PMID- 15796621 TI - Spelt (Triticum aestivum ssp. spelta) as a source of breadmaking flours and bran naturally enriched in oleic acid and minerals but not phytic acid. AB - The nutritional value of breadmaking cereal spelt (Triticum aestivum ssp. spelta) is said to be higher than that of common wheat (Triticum aestivum ssp. vulgare), but this traditional view is not substantiated by scientific evidence. In an attempt to clarify this issue, wholemeal and milling fractions (sieved flour, fine bran, and coarse bran) from nine dehulled spelt and five soft winter wheat samples were compared with regard to their lipid, fatty acid, and mineral contents. In addition, tocopherol (a biochemical marker of germ) was measured in all wholemeals, whereas phytic acid and phosphorus levels were determined in fine bran and coarse bran samples after 1 month of storage. Results showed that, on average, spelt wholemeals and milling fractions were higher in lipids and unsaturated fatty acids as compared to wheat, whereas tocopherol content was lower in spelt, suggesting that the higher lipid content of spelt may not be related to a higher germ proportion. Although milling fractionation produced similar proportions of flour and brans in spelt and wheat, it was found that ash, copper, iron, zinc, magnesium, and phosphorus contents were higher in spelt samples, especially in aleurone-rich fine bran and in coarse bran. Even though phosphorus content was higher in spelt than in wheat brans, phytic acid content showed the opposite trend and was 40% lower in spelt versus wheat fine bran, which may suggest that spelt has either a higher endogenous phytase activity or a lower phytic acid content than wheat. The results of this study give important indications on the real nutritional value of spelt compared to wheat. Moreover, they show that the Ca/Fe ratio, combined with that of oleate/palmitate, provides a highly discriminating tool to authenticate spelt from wheat flours and to face the growing issue of spelt flour adulteration. Finally, they suggest that aleurone differences, the nature of which still needs to be investigated, may account for the differential nutrient composition of spelt and wheat. PMID- 15796622 TI - Different polyphenolic components of soft fruits inhibit alpha-amylase and alpha glucosidase. AB - Polyphenol-rich extracts from soft fruits were tested for their ability to inhibit alpha-amylase and alpha-glucosidase. All extracts tested caused some inhibition of alpha-amylase, but there was a 10-fold difference between the least and most effective extracts. Strawberry and raspberry extracts were more effective alpha-amylase inhibitors than blueberry, blackcurrant, or red cabbage. Conversely, alpha-glucosidase was more readily inhibited by blueberry and blackcurrant extracts. The extent of inhibition of alpha-glucosidase was related to their anthocyanin content. For example, blueberry and blackcurrant extracts, which have the highest anthocyanin content, were the most effective inhibitors of alpha-glucosidase. The extracts most effective in inhibiting alpha-amylase (strawberry and raspberry) contain appreciable amounts of soluble tannins. Other tannin-rich extracts (red grape, red wine, and green tea) were also effective inhibitors of alpha-amylase. Indeed, removing tannins from strawberry extracts with gelatin also removed inhibition. Fractionation of raspberry extracts on Sephadex LH-20 produced an unbound fraction enriched in anthocyanins and a bound fraction enriched in tannin-like polyphenols. The unbound anthocyanin-enriched fraction was more effective against alpha-glucosidase than the original extract, whereas the alpha-amylase inhibitors were concentrated in the bound fraction. The LH-20 bound sample was separated by preparative HPLC, and fractions were assayed for inhibition of alpha-amylase. The inhibitory components were identified as ellagitannins using LC-MS-MS. This study suggests that different polyphenolic components of fruits may influence different steps in starch digestion in a synergistic manner. PMID- 15796623 TI - Red wine polyphenolics suppress the secretion of ApoB48 from human intestinal CaCo-2 cells. AB - Epidemiological studies suggest that the consumption of red wine lowers the risk of cardiovascular disease. Although the cardioprotective effect of red wine has been attributed to its polyphenolic content, presently, very little is known about the mechanisms by which these compounds benefit the cardiovascular system. Therefore, the aim of this study was to elucidate whether red wine polyphenolics attenuate the synthesis and secretion of proatherogenic chylomicrons from intestinal cells. Apolipoprotein B48 levels (marker of intestinal chylomicrons), quantitated by western blotting, were significantly reduced by 30% in cultured CaCo-2 cells and medium when cells were incubated with either dealcoholized red wine, alcoholized red wine, or atorvastatin compared with controls. Intracellular cholesterol availability was also attenuated in cells incubated with dealcoholized red wine (72.5%), alcoholized red wine (81.5%), and atorvastatin (83.5%) compared to control cells. Collectively, this study suggests that red wine polyphenolics downregulate the production of atherogenic chylomicrons from intestinal cells, which may explain the reduced CVD mortality rates following its consumption. PMID- 15796624 TI - Differential transfer of organic micropollutants through intestinal barrier using Caco-2 cell line. AB - Food seems to be one of the main ways of animal and human contamination with polycyclic aromatic hydrocarbons (PAHs) and dioxins. In vivo studies showed a blood absorption of these xenobiotics after their ingestion. Our work aimed at studying the in vitro transfer of PAHs and dioxins through intestinal barrier. Caco-2 cells were cultivated on permeable filters to measure transepithelial permeability of (14)C labeled 2,3,7,8-tetrachlorodibenzo-p-dioxin, benzo[a]pyrene, pyrene, and phenanthrene, which differed in their physicochemical properties. The results showed that the molecules were able to cross intestinal cell layers. All the molecules were detected associated with cells, even if the dioxin was the less uptaken compound. Phenanthrene appeared in basal media faster, and its level after a 6-h exposure was respectively 1.1, 2, and 7 times higher than pyrene, benzo[a]pyrene, and 2,3,7,8-tetrachlorodibenzo-p-dioxin levels. These findings suggest that intestinal epithelium plays a key role in selective permeability and then in bioavailibility of micropollutants. PMID- 15796626 TI - Psychological and behavioral risk factors for obesity onset in adolescent girls: a prospective study. AB - Because little is known about risk factors for obesity, the authors tested whether certain psychological and behavioral variables predicted future onset of obesity. The authors used data from a prospective study of 496 adolescent girls who completed a baseline assessment at age 11-15 years and 4 annual follow-ups. Self-reported dietary restraint, radical weight-control behaviors, depressive symptoms, and perceived parental obesity--but not high-fat food consumption, binge eating, or exercise frequency-predicted obesity onset. Results provide support for certain etiologic theories of obesity, including the affect regulation model. The fact that self-reported, weight-control behaviors identified girls at risk for obesity implies that high-risk youths are not engaging in effective weight-control methods and suggests the need to promote more effective strategies. PMID- 15796627 TI - The association between patient characteristics and the therapeutic alliance in cognitive-behavioral and interpersonal therapy for bulimia nervosa. AB - The therapeutic alliance is an established predictor of psychotherapy outcome. However, alliance research in the treatment of eating disorders has been scant, with even less attention paid to correlates of alliance development. The goal of this study was to examine the relation between specific patient characteristics and the development of the alliance in 2 different treatments for bulimia nervosa (BN). Data derive from a large, randomized clinical trial comparing cognitive- behavioral therapy (CBT) and interpersonal therapy (IPT) for BN. Across both treatments, patient expectation of improvement was positively associated with early- and middle-treatment alliance quality. In CBT, baseline symptom severity was negatively related to middle alliance. In IPT, more baseline interpersonal problems were associated with poorer alliance quality at midtreatment. PMID- 15796629 TI - Adolescent adjustment before and after HIV-related parental death. AB - The impact of HIV-related parental death on 414 adolescents was examined over a period of 6 years. The adjustment of bereaved adolescents was compared over 4 time periods relative to parental death and was also compared with the adjustment of nonbereaved adolescents. Bereaved adolescents had significantly more emotional distress, negative life events, and contact with the criminal justice system than nonbereaved youths; these behaviors did not remain significantly higher after parental death. Depressive symptoms and passive problem solving increased soon after parental death, as compared with nonbereaved adolescents. One year subsequent to parental death, depression and passive problem solving were similar to the levels of nonbereaved peers. Only sexual risk behaviors increased following parental death. These results suggest the importance of early family intervention soon after parental HIV diagnosis, prior to parental death, and sustained over time. PMID- 15796628 TI - Positive affect as a source of resilience for women in chronic pain. AB - A sample of 124 women with osteoarthritis or fibromyalgia, or both, completed initial assessments for demographic data, health status, and personality traits and 10-12 weekly interviews regarding pain, stress, negative affect, and positive affect. Multilevel modeling analyses indicated that weekly elevations of pain and stress predicted increases in negative affect. Both higher weekly positive affect as well as greater positive affect on average resulted in lower negative affect both directly and in interaction with pain and stress. Finally, increases in weekly negative affect and higher average negative affect related to greater levels of pain in subsequent weeks. In contrast, higher levels of overall positive affect predicted lower levels of pain in subsequent weeks. PMID- 15796630 TI - Variation in the drinking trajectories of freshmen college students. AB - F. K. Del Boca, J. Darkes, P. E. Greenbaum, and M. S. Goldman (2004) examined temporal variations in drinking during the freshmen college year and the relationship of several risk factors to these variations. Here, using the same data, the authors investigate whether a single growth curve adequately characterizes the variability in individual drinking trajectories. Latent growth mixture modeling identified 5 drinking trajectory classes: light-stable, light stable plus high holiday, medium-increasing, highdecreasing, and heavy-stable. In multivariate predictor analyses, gender (i.e., more women) and lower alcohol expectancies distinguished the light-stable class from other trajectories; only expectancies differentiated the high-decreasing from the heavy-stable and medium increasing classes. These findings allow for improved identification of individuals at risk for developing problematic trajectories and for development of interventions tailored to specific drinker classes. PMID- 15796631 TI - The occurrence of male-to-female intimate partner violence on days of men's drinking: the moderating effects of antisocial personality disorder. AB - In this study, the moderating effects of antisocial personality disorder (ASPD) on the day-to-day relationship between male partner alcohol consumption and male to-female intimate partner violence (IPV) for men entering a domestic violence treatment program (n=170) or an alcoholism treatment program (n=169) were examined. For both samples, alcohol consumption was associated with an increased likelihood of nonsevere IPV among men without a diagnosis of ASPD but not among men with ASPD (who tended to engage in nonsevere IPV whether they did or did not drink). Drinking was more strongly associated with a likelihood of severe IPV among men with ASPD compared with those without ASPD who also drank. These results provide partial support for a multiple threshold model of intoxication and aggression. PMID- 15796632 TI - Coping skills training and 12-step facilitation for women whose partner has alcoholism: effects on depression, the partner's drinking, and partner physical violence. AB - Women (N=171), distressed from their partners' untreated alcoholism, received either coping skills training (CST), 12-step facilitation (TSF), or delayed treatment (DTC). CST and TSF resulted in lower depression levels than DTC but did not differ from one another. Skill acquisition mediated the treatment effects of CST; Al-Anon attendance did not mediate the TSF effect. Lower depression levels were maintained at 12 months with no differences between groups. Partner drinking decreased from pretreatment to follow-up in the CST and TSF conditions. However, for partners with a history of relationship violence, drinking improved across follow-up in the CST condition but worsened in the TSF condition. Partner relationship violence was less in the CST condition. CST may be particularly useful for women experiencing physical violence from a partner with alcoholism. PMID- 15796633 TI - Less directiveness by therapists improves drinking outcomes of reactant clients in alcoholism treatment. AB - In this study, the authors examined the impact of the interaction between clients' trait reactance and therapists' directiveness on the effectiveness of psychotherapy treatment for alcoholism. Ratings of videotaped treatment sessions were used to measure clients' reactance (N=141) and therapists' directiveness. Models tested for the interaction as a predictor of 1-year posttreatment drinking quantity and frequency. Results indicate that directiveness had a negative impact on outcomes for clients at medium and high levels of reactance but did not affect drinking among clients low in reactance. Increased therapist use of interpretation, confrontation, and introduction of topics was most predictive of more frequent and larger quantities of drinking among reactant clients. This study suggests that research on treatment process can yield significant theoretical and clinical benefits. PMID- 15796634 TI - Recent advances in psychosocial oncology. AB - The April 2005 issue of Journal of Consulting and Clinical Psychology presents 5 articles that address the cancer experience. In this commentary, Vicki S. Helgeson examines how the authors of these articles advance the field in important ways, in particular by using theory-based research, identifying mechanisms of change, identifying moderators of treatment effectiveness, incorporating partners in studies, moving beyond self-report measures of distress, using longer term follow-ups, extending studies to more diverse samples, focusing on sexuality, and using longitudinal designs. Helgeson concludes by identifying several challenges that researchers in this area face and by highlighting future avenues for research. PMID- 15796635 TI - Using problem-solving skills training to reduce negative affectivity in mothers of children with newly diagnosed cancer: report of a multisite randomized trial. AB - Mothers of children with cancer experience significant distress associated with their children's diagnosis and treatment. The efficacy of problem-solving skills training (PSST), a cognitive-behavioral intervention based on problem-solving therapy, was assessed among 430 English- and Spanish-speaking mothers of recently diagnosed patients. Participants were randomized to usual psychosocial care (UPC; n=213) or UPC plus 8 sessions of PSST (PSST; n=217). Compared with UPC mothers, PSST mothers reported significantly enhanced problem-solving skills and significantly decreased negative affectivity. Although effects were largest immediately after PSST, several differences in problem-solving skills and distress levels persisted to the 3-month follow-up. In general, efficacy for Spanish-speaking mothers exceeded that for English-speaking mothers. Findings also suggest young, single mothers profit most from PSST. PMID- 15796636 TI - The effects of personal construct group therapy on breast cancer survivors. AB - In this study, the authors evaluated the effects of a brief personal construct group therapy on breast cancer survivors (N=42) randomly assigned to either the treatment or wait-list control condition. The Gottschalk Gleser Content Analysis Scales were used to measure the effects for group across time (pre- and posttreatment, pretreatment, and 3-month posttreatment) on the personal construct states of threat, threat to existence, dislocation, and hope. Analyses showed that the beneficial effects of therapy achieved posttreatment were maintained at 3-month follow-up. The therapeutic group factors (I. D. Yalom, 1995) identified by the therapy group members as helpful in achieving these outcomes are also reported. PMID- 15796637 TI - A prospective study of autobiographical memory and posttraumatic stress disorder following cancer. AB - In this study, the authors investigated the relationship between autobiographical memory and the onset and maintenance of distressing memories following cancer. In Study 1, participants recently diagnosed with head, neck, or lung cancer were assessed for acute stress disorder (ASD). Participants with ASD reported fewer specific memories than did participants without ASD. In Study 2, the same participants were assessed 6 months later for autobiographical memory and cancer related posttraumatic stress disorder (PTSD). Deficits in the retrieval of specific memories in Study 1 were not predictive of subsequent PTSD. Increased hopelessness during the 6 months was associated with a decline in the retrieval of positive memories and an increase in the retrieval of negative memories. These findings accord with propositions that retrieval of distressing memories is guided by current self-image and attitude toward one's future. PMID- 15796638 TI - Perceived partner reactions to diagnosis and treatment of breast cancer: impact on psychosocial and psychosexual adjustment. AB - Two studies examined breast cancer patients' perceptions of their partners' reactions to their diagnosis and treatment as influences on 3 aspects of patients' well-being: psychosexual adjustment, emotional distress, and marital satisfaction. Study 1, cross-sectional, indicated that partner initiation of sex, frequency of sex, a positive 1st sexual experience after treatment, and especially perception of the partner's emotional involvement in the relationship, were relevant to these outcomes. Study 2, longitudinal, confirmed many of these findings in prospective tests across 1 year of recovery after surgery. Partner involvement prospectively predicted all 3 outcomes. Partner initiation of sex predicted greater marital satisfaction; partner adverse reaction to the scar predicted less marital satisfaction. Rated quality of the 1st sexual experience after treatment predicted less distress. The pattern suggests that women's impressions of their partners' emotional involvement after surgery for breast cancer forecast their adjustment in sexual, marital, and emotional arenas over the following year. PMID- 15796639 TI - Stages of change in treatment-seeking pathological gamblers. AB - The transtheoretical model has been applied to many addictive disorders. In this study, psychometrics properties of the University of Rhode Island Change Assessment (URICA) scale were evaluated in 234 pathological gamblers initiating treatment. Four components were identified--reflective of precontemplation, contemplation, action, and maintenance stages--with internal consistency from .74 to .88. Cluster analyses identified 4 patterns of responding, ranging from ambivalent to active change. The 4 clusters differed with respect to baseline gambling variables and treatment engagement and outcomes assessed 2 months later. A continuous measure of readiness to change was also correlated with gambling severity and predictive of reductions in gambling. This study provides initial support for reliability and validity of the URICA in treatment-seeking gamblers, and it suggests that stage of change may have an impact on outcomes. PMID- 15796640 TI - The therapy process observational coding system-alliance scale: measure characteristics and prediction of outcome in usual clinical practice. AB - The authors describe psychometric characteristics of the new Therapy Process Observational Coding System-Alliance scale (TPOCS-A; B. D. McLeod, 2001) and illustrate its use in the study of treatment as usual. The TPOCS-A uses session observation to assess child-therapist and parent-therapist alliance. Both child and parent forms showed acceptable interrater reliability and internal consistency; when applied to cases treated for internalizing disorders, both forms were associated with youth outcomes. Child-therapist alliance during treatment predicted reduced anxiety symptoms at the end of treatment. Parent therapist alliance during treatment predicted reduced internalizing, anxiety, and depression symptoms at the end of treatment. The findings held up well after confounding variables were controlled, which suggests that both child-therapist and parent-therapist alliance play key (and potentially different) roles in the outcome of treatment as usual. PMID- 15796642 TI - Involving support partners in obesity treatment. AB - In this study, the authors examined whether the number or success of weight loss partners influences participants' outcomes in behavioral weight loss treatment. Overweight participants (n=109) assigned to an exercise intensive group in a larger trial were encouraged to invite up to 3 partners to attend treatment. Weight losses at 6, 12, and 18 months were not associated with the number of partners (0-3) but were associated with the weight loss success of the partners. Participants with at least 1 successful partner (weight loss>or=10% at 6 months) lost significantly (p=.004) more weight at 6, 12, and 18 months than those with no successful partners and those without partners. Interclass correlations of weight change between participants and their partner(s) were strong at all time points (ps<.01). Support partners appear to only be beneficial in obesity treatment when partners themselves lose weight. PMID- 15796641 TI - The additive benefit of hypnosis and cognitive-behavioral therapy in treating acute stress disorder. AB - This research represents the first controlled treatment study of hypnosis and cognitive- behavioral therapy (CBT) of acute stress disorder (ASD). Civilian trauma survivors (N=87) who met criteria for ASD were randomly allocated to 6 sessions of CBT, CBT combined with hypnosis (CBT-hypnosis), or supportive counseling (SC). CBT comprised exposure, cognitive restructuring, and anxiety management. CBT-hypnosis comprised the CBT components with each imaginal exposure preceded by a hypnotic induction and suggestions to engage fully in the exposure. In terms of treatment completers (n=69), fewer participants in the CBT and CBT hypnosis groups met criteria for posttraumatic stress disorder at posttreatment and 6-month follow-up than those in the SC group. CBT-hypnosis resulted in greater reduction in reexperiencing symptoms at posttreatment than CBT. These findings suggest that hypnosis may have use in facilitating the treatment effects of CBT for posttraumatic stress. PMID- 15796643 TI - Fear of severe acute respiratory syndrome (SARS) among health care workers. AB - In this study, the authors examined fear related to severe acute respiratory syndrome (SARS) among 2 samples of hospital staff in Hong Kong. Sample 1 included health care workers (n=82) and was assessed during the peak of the SARS epidemic. Sample 2 included hospital staff who recovered from SARS (n=97). The results show that participants in both samples had equal, if not more, concern about infecting others (especially family members) than being self-infected. Sample 1 participants had stronger fear related to infection than Sample 2 participants, who seemed to be concerned more about other health problems and discrimination. Participants with lower self-efficacy tended to have higher fear related to SARS. Fear related to SARS was also correlated positively with posttraumatic stress symptoms among respondents of Sample 2 (recovered staff). Interventions based on these findings are described. PMID- 15796644 TI - Assigned versus perceived placebo effects in nicotine replacement therapy for smoking reduction in Swiss smokers. AB - In this report, the authors explore the relationships of perceived treatment to outcome in a large, placebo-controlled trial of nicotine replacement treatment for smoking reduction. In the original study (J. F. Etter, E. Laszlo, J. P. Zellweger, C. Perrot, & T. V. Perneger, 2002), which was conducted in French speaking Switzerland, smokers were randomly assigned to receive nicotine, matching placebo products, or no intervention. At the end of the 6-month study, participants were asked to guess whether they had received nicotine or placebo. In the present analysis, the authors examined the difference in smoking reduction between those who believed they had received nicotine and those who believed they had received placebo. Regardless of actual treatment, smokers who believed they had received nicotine had significantly better outcome than those who believed they had received placebo. PMID- 15796646 TI - The relationship between acute stress disorder and posttraumatic stress disorder following cancer. AB - In this study, the authors investigated the relationship between acute stress disorder (ASD) and posttraumatic stress disorder (PTSD) following cancer diagnosis. Patients who were recently diagnosed with 1st onset head and neck or lung malignancy (N=82) were assessed for ASD within the initial month following their diagnosis and reassessed (n=63) for PTSD 6 months following their cancer diagnosis. At the initial assessment, 28% of patients had ASD, and 32% displayed subsyndromal ASD. At follow-up, PTSD was diagnosed in 53% of patients who had been diagnosed with ASD and in 11% of those who had not met criteria for ASD; 36% of patients with PTSD did not initially display ASD. In this study, the authors question the use of the ASD diagnosis to identify recently diagnosed patients at risk of PTSD. PMID- 15796645 TI - Prize reinforcement contingency management for cocaine dependence: integration with group therapy in a methadone clinic. AB - In this study, the authors evaluated a low-cost contingency management (CM) procedure for reducing cocaine use and enhancing group therapy attendance in 77 cocaine-dependent methadone patients. Patients were randomly assigned to 12 weeks of standard treatment or standard treatment with CM, in which patients earned the opportunity to win prizes ranging from $1 to $100 for submitting cocainenegative samples and attending therapy. Patients in the CM condition submitted more cocaine-negative samples and attended more groups than patients in standard treatment. The best predictor of cocaine abstinence at follow-up was duration of abstinence during treatment. On average, patients in the CM condition earned $117 in prizes. Data from this study suggest that some aspects of reinforcement can be implemented in group therapy in community-based clinics. PMID- 15796647 TI - Methodology in family science: introduction to the special issue. AB - Methodology influences every facet of scientific inquiry including the processes of framing questions, operationalizing constructs, sampling individuals within populations and observations within persons, analyzing data, and bringing meaning to the results. This introduction to the special issue on methodology emphasizes the recursive interaction of research methodology and theories underlying family science. Because the science of family psychology is richly complex, so too must the methods for examining couple and family phenomena be equally diverse. Articles comprising this special issue vary in their emphasis on research design versus data-analytic techniques and on confirmatory versus discovery-oriented strategies. The special issue is intended to advance not only the conduct of family science but also the understanding and integration of an increasingly complex research literature by those engaged in education and clinical interventions. ((c) 2005 APA, all rights reserved). PMID- 15796649 TI - Change process research in couple and family therapy: methodological challenges and opportunities. AB - In our field, the study of therapeutic change processes lags behind the study of treatment efficacy. Nonetheless, in the past 10 years major strides have been made in delineating change process mechanisms in couple and family therapy. To focus the efforts of future change process researchers, the authors discuss 5 critical needs: (a) more well-articulated, midrange theory about systemic change processes; (b) more attention to client change processes; (c) more attention to covert intrapersonal processes (emotion, cognition, and clients' experience of the alliance); (d) better articulation of strategies for analyzing data from multiple participants; and (e) more focus on the degree to which various change processes work similarly (or not) for diverse couples and families. PMID- 15796648 TI - Methodologically sound, cost-effective research on the outcome of couple therapy. AB - Because of its potential to answer pressing applied questions about what treatments to use with which couples as well as its potential to answer basic questions about couple functioning, outcome research on couple therapy is vital to the field. This article describes the primary methodological issues that an investigator faces in conducting outcome research during treatment development, efficacy testing, and effectiveness evaluation. Descriptions of what has been done in the past are integrated with recommendations about what should be done in the future. Mindful of the potential expense of outcome research, the authors suggest ways in which questions can be investigated cost effectively. PMID- 15796650 TI - Assessing the costs, benefits, cost-benefit ratio, and cost-effectiveness of marital and family treatments: why we should and how we can. AB - The vast majority of outcome studies examining the effects of marital and family treatments focus exclusively on indicators of and changes in familial functioning and individual members' psychosocial adjustment, but fail to measure, report, or analyze treatment costs, benefits, cost-benefit ratio, or cost-effectiveness. Because of growing concerns about spiraling health care costs, clinical and economic outcomes constitute equally important and complementary aspects of any evaluation of marital and family treatments. The twofold purpose of this article is to define different components of cost analyses of health-related interventions, including marital and family treatments, and to describe methods for calculating and integrating clinical and cost outcome information when evaluating marital and family treatments. There are significant opportunities to promote the use of such treatments by conducting and reporting the results of cost analyses. PMID- 15796651 TI - Qualitative research and family psychology. AB - Qualitative approaches have much to offer family psychology. Among the uses for qualitative methods are theory building, model and hypothesis testing, descriptions of lived experiences, typologies, items for surveys and measurement tools, and case examples that answer questions that surveys cannot. Despite the usefulness of these products, issues related to generalizability, subjectivity, and language, among others, block some researchers from appreciating the contributions that qualitative methods can make. This article provides descriptions of procedures that lead to these useful products and discusses alternative ways of understanding aspects of qualitative approaches that some researchers view as problematic. PMID- 15796652 TI - Narratives in and about families: an examination of coding schemes and a guide for family researchers. AB - This article aims to provide the reader with a general overview of narrative methods as they apply to the study of family relationships. How narratives are collected and issues pertaining to training raters are considered. Scoring systems have been developed that code for thematic content, relative coherence, and co-constructive processes. Validity for these methods is examined through (a) concordance between narrative codes and directly observed behavior within similar domains, (b) links between narratives and self-report measures, and (c) the relation between narratives and psychological functioning. A guide for researchers in their consideration of adopting narrative methods is offered. The article concludes with a consideration of linking different coding schemes and challenges for future researchers. PMID- 15796653 TI - Video-recall procedures for examining subjective understanding in observational data. AB - Video-recall procedures allow researchers and clinicians to ascertain participants' subjective understanding of their interactions with significant others. The procedures involve recording participants' interactions and subsequently asking them to review the recorded interactions. The reviewing involves participants providing feedback about their perceptions of their interactions, behaviors, or experiences during the original interactions. Participants' subjective understandings can be contrasted with each other and with those of trained coders to examine shared and discrepant understandings and the differential predictive utility of interactional processes. In this article, the authors briefly review the literature on video-recall procedures, discuss their potential for application in a variety of areas in family psychology, and provide an analysis of current video-recall procedures. Finally, the authors present the Digital Video-Recall System as an example of current video-recall methodology. PMID- 15796654 TI - Multilevel methods for modeling observed sequences of family interaction. AB - Observation of interaction plays a central role in family research. This article discusses how to analyze sequential data generated by discrete microcoding methods to test hypotheses about family interaction. Current methods for studying sequential data are presented, and their limits are discussed. Building on recent applications of contingency table analysis to such data, a multilevel log-linear model is presented that can specify and estimate indicators of individual behavioral tendencies and antecedent-consequent relationships among behaviors, both within and across samples of families. An example of this method is presented using data from a study of couples facing job loss. Potential extensions of this framework for future research are discussed. PMID- 15796655 TI - Using diary methods to study marital and family processes. AB - Diary methods allow researchers to study marital and family processes within the context of daily life in a way that is not possible with more traditional methods. The authors review applications of diary designs in marital and family research and detail the types of research questions that can uniquely be asked of dyadic/family diary data. Technological developments for the use of electronic palm-top devices for implementing diary methods are also reviewed. Additionally, the authors discuss specific issues relevant to the analysis of diary data that come from dyads or families. Last, the authors raise unresolved issues and directions for future research in the use of diary methods for studying marital and family processes. PMID- 15796656 TI - Using multilevel models to analyze couple and family treatment data: basic and advanced issues. AB - Couple and family treatment data present particular challenges to statistical analyses. Partners and family members tend to be more similar to one another than to other individuals, which raises interesting possibilities in the data analysis but also causes significant problems with classical, statistical methods. The present article presents multilevel models (also called hierarchical linear models, mixed-effects models, or random coefficient models) as a flexible analytic approach to couple and family longitudinal data. The article reviews basic properties of multilevel models but focuses primarily on 3 important extensions: missing data, power and sample size, and alternative representations of couple data. Information is presented as a tutorial, with a Web appendix providing datasets with SPSS and R code to reproduce the examples. PMID- 15796657 TI - Designing, testing, and interpreting interactions and moderator effects in family research. AB - This article is a primer on issues in designing, testing, and interpreting interaction or moderator effects in research on family psychology. The first section focuses on procedures for testing and interpreting simple effects and interactions, as well as common errors in testing moderators (e.g., testing differences among subgroup correlations, omitting components of products, and using median splits). The second section, devoted to difficulties in detecting interactions, covers such topics as statistical power, measurement error, distribution of variables, and mathematical constraints of ordinal interactions. The third section, devoted to design issues, focuses on recommendations such as including reliable measures, enhancing statistical power, and oversampling extreme scores. The topics covered should aid understanding of existing moderator research as well as improve future research on interaction effects. PMID- 15796658 TI - Cluster analysis in family psychology research. AB - This article discusses the use of cluster analysis in family psychology research. It provides an overview of potential clustering methods, the steps involved in cluster analysis, hierarchical and nonhierarchical clustering methods, and validation and interpretation of cluster solutions. The article also reviews 5 uses of clustering in family psychology research: (a) deriving family types, (b) studying families over time, (c) as an interface between qualitative and quantitative methods, (d) as an alternative to multivariate interactions in linear models, and (e) as a data reduction technique for small samples. The article concludes with some cautions for using clustering in family psychology research. PMID- 15796659 TI - Analyzing nonindependent outcomes in couple therapy using the actor-partner interdependence model. AB - The nonindependent components of couple data require data-analytic strategies tailored to the interpersonal processes occurring between relationship partners. This article examines the benefits of a specifically dyadic analytic method, the actor-partner interdependence model (APIM), for evaluating interpartner influence across time. Both conceptual and methodological features of the APIM are exemplified by applying this model to observations of negative and positive affect and global distress in spouses participating in a randomized trial of couple therapy. In addition to elucidating specific advantages of the APIM relative to alternative data-analytic strategies, the current results shed new light on previous findings from a comparative treatment outcome study evaluating behavioral and insight-oriented approaches to couple therapy. PMID- 15796660 TI - Survival analysis in family research. AB - Within the field of family psychology, questions regarding the risk of event occurrence may be common. Such questions, about whether and when events occur and what predicts these occurrences, pose particular methodological challenges and are often best addressed via a statistical method known as survival analysis. This article provides a brief overview of that method, explicating through a data example the major components of a discrete-time survival analysis. Readers not familiar with this method are encouraged to use this article as an introduction to survival analysis and recognize its potential usefulness within the field of family psychology. PMID- 15796661 TI - Ethical issues in couple and family research. AB - Federal regulations, ethical standards, and state laws governing ethics do not adequately address important issues in couple and family research. Including multiple family members, particularly dependent minors, in research requires the special application of fundamental ethical issues, such as confidentiality, privacy, and informed consent. The sensitive, commingled nature of couple and family information necessitates clear policies about data ownership and disclosure. Researchers need to have respect for the family as a unit and to evaluate benefits versus harms for the family as well as for individuals. This article highlights areas of potential concern and ambiguity related to abuse reporting and Certificates of Confidentiality and also addresses ethical issues with observational data, intervention studies, longitudinal designs, and computer assisted research. PMID- 15796662 TI - Social exclusion impairs self-regulation. AB - Six experiments showed that being excluded or rejected caused decrements in self regulation. In Experiment 1, participants who were led to anticipate a lonely future life were less able to make themselves consume a healthy but bad-tasting beverage. In Experiment 2, some participants were told that no one else in their group wanted to work with them, and these participants later ate more cookies than other participants. In Experiment 3, excluded participants quit sooner on a frustrating task. In Experiments 4-6, exclusion led to impairment of attention regulation as measured with a dichotic listening task. Experiments 5 and 6 further showed that decrements in self-regulation can be eliminated by offering a cash incentive or increasing self-awareness. Thus, rejected people are capable of self-regulation but are normally disinclined to make the effort. PMID- 15796663 TI - Incorporating if...then...personality signatures in person perception: beyond the person-situation dichotomy. AB - Three studies investigated conditions in which perceivers view dispositions and situations as interactive, rather than independent, causal forces when making judgments about another's personality. Study 1 showed that perceivers associated 5 common trait terms (e.g., friendly and shy) with characteristic if...then... (if situation a, then the person does x, but if situation b, then the person does y) personality signatures. Study 2 demonstrated that perceivers used information about a target's stable if...then... signature to infer the target's motives and traits; dispositional judgments were mediated by inferences about the target's motivations. Study 3 tested whether perceivers draw on if...then... signatures when making judgments about Big Five trait dimensions. Together, the findings indicate that perceivers take account of person-situation interactions (reflected in if...then... signatures) in everyday explanations of social behavior and personality dispositions. Boundary conditions are also discussed. PMID- 15796664 TI - Differential access to information and anticipated group interaction: impact on individual reasoning. AB - Two studies examined the impact of relative differences in access to information and anticipated group interaction on individual reasoning. On 2 different reasoning tasks (P. C. Wason's [1966] selection task and D. Kahneman & A. Tversky's [1973] lawyer-engineer problem), participants sensing that they knew more in anticipation of group interaction or knew less when not anticipating interaction were less susceptible to typical cognitive biases demonstrated by these tasks. Study 2 also showed that the effect of these social contexts was contingent on the task presentation format. Thus, knowing more in anticipation of group interaction and knowing less when not anticipating group interaction seemingly compensated for task features that enhance suboptimal reasoning strategies. These results illustrate the importance of the social context in which reasoning is situated and are discussed in terms of cognitive tuning, social comparison, and social motivations. PMID- 15796665 TI - Self-regulation and self-presentation: regulatory resource depletion impairs impression management and effortful self-presentation depletes regulatory resources. AB - Self-presentation may require self-regulation, especially when familiar or dispositional tendencies must be overridden in service of the desired impression. Studies 1-4 showed that self-presentation under challenging conditions or according to counter-normative patterns (presenting oneself modestly to strangers, boastfully to friends, contrary to gender norms, to a skeptical audience, or while being a racial token) led to impaired self-regulation later, suggesting that those self-presentations depleted self-regulatory resources. When self-presentation conformed to familiar, normative, or dispositional patterns, self-regulation was less implicated. Studies 5-8 showed that when resources for self-regulation had been depleted by prior acts of self-control, self presentation drifted toward less-effective patterns (talking too much, overly or insufficiently intimate disclosures, or egotistical arrogance). Thus, inner processes may serve interpersonal functions, although optimal interpersonal activity exacts a short-term cost. PMID- 15796666 TI - Power and the creation of patronizing environments: the stereotype-based behaviors of the powerful and their effects on female performance in masculine domains. AB - This work tested the following hypothesis: When powerful men stereotype their female subordinates in masculine domains, they behave in patronizing ways that affect the performance of their subordinates. Experiment 1 examined the stereotyping tendencies and patronizing behaviors of the powerful. Findings revealed that powerful men who stereotyped their female subordinates (i.e., those who were weakness focused) gave female subordinates few valued resources but much praise. In Experiment 2, low-power participants received resources (valued or devalued positions) and praise (high or low) from a powerful man. Subordinates who were assigned to a devalued position but received high praise (i.e., the patronizing behavior mirrored from Experiment 1) were angry. However, men performed better in the anger-inspiring situation, whereas women performed worse. PMID- 15796667 TI - Daily depression and cognitions about stress: evidence for a traitlike depressogenic cognitive style and the prediction of depressive symptoms in a prospective daily diary study. AB - The authors examined the stability and dynamic structure of negative cognitions made to naturalistic stressors and the prediction of depressive symptoms in a daily diary study. Young adults reported on dispositional depression vulnerabilities at baseline, including a depressogenic cognitive style, dysfunctional attitudes, rumination, neuroticism, and initial depression, and then completed short diaries recording the inferences they made to the most negative event of the day along with their experience of depressive symptoms every day for 35 consecutive days. Daily cognitions about stressors exhibited moderate stability across time. A traitlike model, rather than a contextual one, explained this pattern of stability best. Hierarchical linear modeling analyses showed that individuals' dispositional depressogenic cognitive style, neuroticism, and their daily negative cognitions about stressors predicted fluctuations in daily depressive symptoms. Dispositional neuroticism and negative cognitive style interacted with daily negative cognitions in different ways to predict daily depressive symptoms. PMID- 15796668 TI - In genes we trust: the biological component of psychological essentialism and its relationship to mechanisms of motivated social cognition. AB - Three studies analyzed the biological component of psychological essentialism (laypeople's belief that social categories have an underlying nature/natural foundation) as it pertains to mechanisms of motivated social cognition. A new scale assessing the belief in genetic determinism is introduced as a measure of the biological component of essentialism. Results speak to the reliability and validity of the scale and show that essentialist beliefs are associated with basic social-cognitive motives and are also related to processes of stereotyping and prejudice. An experimental study found that rendering essentialist information salient elicits increased levels of prejudice and in-group bias, particularly in persons holding chronic essentialist beliefs. PMID- 15796669 TI - You can't always get what you want: educational attainment, agency, and choice. AB - Using educational attainment to indicate socioeconomic status, the authors examined models of agency and effects of choice among European American adults of different educational backgrounds in 3 studies. Whereas college-educated (BA) participants and their preferred cultural products (i.e., rock music lyrics) emphasized expressing uniqueness, controlling environments, and influencing others, less educated (HS) participants and their preferred cultural products (i.e., country music lyrics) emphasized maintaining integrity, adjusting selves, and resisting influence. Reflecting these models of agency, HS and BA participants differently responded to choice in dissonance and reactance paradigms: BA participants liked chosen objects more than unchosen objects, but choice did not affect HS participants' preferences. Results suggest that HS and BA models of agency qualitatively differ, despite overlap between HS and BA worlds. PMID- 15796670 TI - Object-based attention and cognitive tunneling. AB - Simulator-based research has shown that pilots cognitively tunnel their attention on head-up displays (HUDs). Cognitive tunneling has been linked to object-based visual attention on the assumption that HUD symbology is perceptually grouped into an object that is perceived and attended separately from the external scene. The present research strengthens the link between cognitive tunneling and object based attention by showing that (a) elements of a visual display that share a common fate are grouped into a perceptual object and that this grouping is sufficient to sustain object-based attention, (b) object-based attention and thereby cognitive tunneling is affected by strategic focusing of attention, and (c) object-based attention is primarily inhibitory in nature. PMID- 15796671 TI - The case for coherence in scientific explanations: quantitative details can hurt qualitative understanding. AB - In Experiments 1A and 1B, students read a concise booklet containing 653 words and 6 illustrations describing the formation, propagation, and dispersion of ocean waves (concise group) or an expanded booklet containing 327 additional words and 5 additional illustrations describing relevant mathematical formulas and computations interspersed throughout the lesson (expanded group). In Experiment 2, students viewed a multimedia presentation of narrated animations based on the concise or expanded booklet. In both studies, the expanded group performed more poorly than did the concise group on problem-solving transfer tests. The added quantitative details may have distracted the learner from constructing a qualitative model of the process of ocean waves. PMID- 15796672 TI - Visual perception of touchdown point during simulated landing. AB - Experiments examined the accuracy of visual touchdown point perception during oblique descents (1.5 degrees -15 degrees ) toward a ground plane consisting of (a) randomly positioned dots, (b) a runway outline, or (c) a grid. Participants judged whether the perceived touchdown point was above or below a probe that appeared at a random position following each display. Although judgments were unacceptably imprecise and biased for moving dot and runway displays, accurate and unbiased judgments were found for grid displays. It is concluded that optic flow per se does not appear to be sufficient for a pilot to land an airplane and that the systematic errors associated with optic flow under sparse conditions may be responsible for the common occurrence of landing incidents in so-called "black hole" situations. PMID- 15796673 TI - Source recall enhances children's discrimination of seen and heard events. AB - The effects of rehearsing actions by source (slideshow vs. story) and of test modality (picture vs. verbal) on source monitoring were examined. Seven- to 8 year-old children (N = 30) saw a slideshow event and heard a story about a similar event. One to 2 days later, they recalled the events by source (source recall), recalled the events without reference to source (no-source-cue recall), or engaged in no recall. Seven to 8 days later, all children received verbal and picture source-monitoring tests. Children in the source recall group were less likely than children in the other groups to claim they saw actions merely heard in the story. No-source-cue recall impaired source identification of story actions. The picture test enhanced recognition, but not source monitoring, of slide actions. Increasing the distinctiveness of the target events (Experiment 2) allowed the picture test to facilitate slideshow action discrimination by children in the no-recall group. PMID- 15796674 TI - Learning the value of a new currency from prices. AB - In European countries, field studies investigate how citizens acquire knowledge of the new currency, the euro. In 3 laboratory experiments, the authors recruited 168 undergraduates to examine whether such accurate knowledge is acquired from learning prices in the new currency. The results show fast learning of prices of duration of cellular phone calls (quantity) when the prices were proportional to quantity. Inferences of call duration from given prices were likewise found to be accurate. Lower accuracy was however observed for one-to-many mappings of quantity on price and the inverse one-to-many mappings of price on quantity. Price variation may be an important reason for the difficulty in learning the value of the new currency observed in field studies. PMID- 15796675 TI - Perspective-taking judgments among young adults, middle-aged, and elderly people. AB - Perspective-taking judgments among young adults, middle-aged, and elderly people were examined. In 1 condition, participants were instructed to judge the likelihood of acceptance of a painkiller as a function of 3 cues: severity of the condition, potential side effects, and level of trust in the health care provider. In the other condition, participants were instructed to judge the likelihood of purchasing pieces of clothing. Judgments were given from 2 viewpoints: the viewpoint of another person known to place no importance on one of the 3 cues, and the viewpoint of another person known to place very great importance on this cue. The hypotheses were that elderly people would not, to the same extent as young adults, be likely to discount the impact of the "no importance" cue, and to magnify the impact of the "very important" cue. In both judgment situations, the results support these hypotheses. PMID- 15796676 TI - On the psychology of confessions: does innocence put innocents at risk? AB - The Central Park jogger case and other recent exonerations highlight the problem of wrongful convictions, 15% to 25% of which have contained confessions in evidence. Recent research suggests that actual innocence does not protect people across a sequence of pivotal decisions: (a) In preinterrogation interviews, investigators commit false-positive errors, presuming innocent suspects guilty; (b) naively believing in the transparency of their innocence, innocent suspects waive their rights; (c) despite or because of their denials, innocent suspects elicit highly confrontational interrogations; (d) certain commonly used techniques lead suspects to confess to crimes they did not commit; and (e) police and others cannot distinguish between uncorroborated true and false confessions. It appears that innocence puts innocents at risk, that consideration should be given to reforming current practices, and that a policy of videotaping interrogations is a necessary means of protection. PMID- 15796677 TI - The primacy of cognition in schizophrenia. AB - Cognitive tasks and concepts are used increasingly in schizophrenia science and treatment. Recent meta-analyses show that across a spectrum of research domains only cognitive measures distinguish a majority of schizophrenia patients from healthy people. Average effect sizes derived from common clinical tests of attention, memory, language, and reasoning are twice as large as those obtained in structural magnetic resonance imaging and positron emission tomography studies. Chronic stress, genes, brain disturbances, task structure, gender, and sociocultural background may all enhance the sensitivity of cognitive performance to schizophrenia. At the same time, disease heterogeneity and the presence of endophenotypes and subtypes within the patient population may place upper limits on the strength of any specific cognitive finding. Schizophrenia is a complex biobehavioral disorder that manifests itself primarily in cognition. PMID- 15796678 TI - Environmental toxicants and developmental disabilities: a challenge for psychologists. AB - Developmental, learning, and behavioral disabilities are a significant public health problem. Environmental chemicals can interfere with brain development during critical periods, thereby impacting sensory, motor, and cognitive function. Because regulation in the United States is based on limited testing protocols and essentially requires proof of harm rather than proof of lack of harm, some undefined fraction of these disabilities may reflect adverse impacts of this "vast toxicological experiment" (H. L. Needleman, as quoted in B. Weiss & P. J. Landrigan, 2000, p. 373). Yet the hazards of environmental pollutants are inherently preventable. Psychologists can help prevent developmental disabilities by mobilizing and affecting public policy, educating and informing consumers, contributing to interdisciplinary research efforts, and taking action within their own homes and communities to reduce the toxic threat to children. PMID- 15796684 TI - On hardiness and other pathways to resilience. PMID- 15796685 TI - The human capacity for growth through adversity. PMID- 15796686 TI - Has resilience to severe trauma been underestimated? PMID- 15796687 TI - Conceptual clarifications in the study of resilience. PMID- 15796689 TI - Natural resilience and innate mental health. PMID- 15796690 TI - Stereotype threat research and the assumptions underlying analysis of covariance. PMID- 15796691 TI - Stereotype threat might explain the Black-White test-score difference. PMID- 15796692 TI - Stereotype threat and the social and scientific contexts of the race achievement gap. PMID- 15796694 TI - Quantum chaos meets coherent control. AB - Coherent control of atomic and molecular processes has been a rapidly developing field. Applications of coherent control to large and complex molecular systems are expected to encounter the effects of chaos in the underlying classical dynamics, i.e., quantum chaos. Hence, recent work has focused on examining control in model chaotic systems. This work is reviewed, with an emphasis on a variety of new quantum phenomena that are of interest to both areas of quantum chaos and coherent control. PMID- 15796695 TI - Femtosecond laser photoelectron spectroscopy on atoms and small molecules: prototype studies in quantum control. AB - We review prototype studies in the area of quantum control with femtosecond lasers. We restrict this discussion to atoms and diatomics under gas-phase collision-free conditions to allow for a comparison between theory and experiment. Both the perturbative regime and the nonperturbative regime of the light-matter interaction are addressed. To that end, atomic/molecular beam techniques are combined together with femtosecond laser techniques and energy resolved photoelectron spectroscopy and ion detection. Highly detailed information on the laser-induced quantum dynamics is extracted with the help of kinetic energy-resolved photoelectron spectroscopy. PMID- 15796696 TI - Nonstatistical dynamics in thermal reactions of polyatomic molecules. AB - A brief review is presented of post-RRKM models for unimolecular reaction kinetics. The dynamics of the gas-phase S(N)2 reaction are discussed, and the important role of phase-space bottlenecks is highlighted. The remainder of the review is devoted to experimental and trajectory simulation results on thermal reactions of organic molecules that exhibit nonstatistical dynamics quite unlike that seen in the S(N)2 reaction. Specifically, the intermediates generated in these reactions decay much faster than RRKM theory would predict, and often with bimodal or multimodal lifetime distributions. A qualitative model for this behavior based on overlaps of transitional regions in the molecular phase space is discussed. PMID- 15796697 TI - Rydberg wavepackets in molecules: from observation to control. AB - Significant advances in laser technology have led to an increasing interest in the time evolution of Rydberg wavepackets as a means to understanding, and ultimately controlling, quantum phenomena. Rydberg wavepackets in molecules are particularly interesting as they possess many of the dynamical complications of large molecules, such as nonadiabatic coupling between the various degrees of freedom, yet they remain tractable experimentally and theoretically. This review explains in detail how the method of interfering wavepackets can be applied to observe and control Rydberg wavepackets in molecules; it discusses the achievements to date and the possibilities for the future. PMID- 15796698 TI - Electron injection at dye-sensitized semiconductor electrodes. AB - Electron injection at dye-sensitized semiconductors is reviewed. Particular emphasis is placed on theoretical and photoelectrochemical studies of dye sensitized planar and single-crystal electrodes. The accepted mechanism of electron injection, which was derived from these classical studies, is introduced. Selected photoelectrochemical studies of dye-sensitized nanocrystalline semiconductors are reviewed; emphasis is given to factors that influence the efficiencies of electron injection and charge recombination. The development of quasi-solid-state nanocrystalline dye-sensitized solar cells is also discussed. Recent time-resolved spectroscopic studies of electron injection and charge recombination are reviewed. These studies have led to a better understanding of electron injection mechanisms, and have revealed the limitations of the classical models. PMID- 15796699 TI - Quantum mode-coupling theory: formulation and applications to normal and supercooled quantum liquids. AB - We review our recent efforts to formulate and study a mode-coupling approach to real-time dynamic fluctuations in quantum liquids. Comparison is made between the theory and recent neutron scattering experiments performed on liquid ortho deuterium and para-hydrogen. We discuss extensions of the theory to supercooled and glassy states where quantum fluctuations compete with thermal fluctuations. Experimental scenarios for quantum glassy liquids are briefly discussed. PMID- 15796700 TI - Quantum mechanics of dissipative systems. AB - Quantum dissipation involves both energy relaxation and decoherence, leading toward quantum thermal equilibrium. There are several theoretical prescriptions of quantum dissipation but none of them is simple enough to be treated exactly in real applications. As a result, formulations in different prescriptions are practically used with different approximation schemes. This review examines both theoretical and application aspects on various perturbative formulations, especially those that are exact up to second-order but nonequivalent in high order system-bath coupling contributions. Discrimination is made in favor of an unconventional formulation that in a sense combines the merits of both the conventional time-local and memory-kernel prescriptions, where the latter is least favorite in terms of the applicability range of parameters for system-bath coupling, non-Markovian, and temperature. Also highlighted is the importance of correlated driving and dissipation effects, not only on the dynamics under strong external field driving, but also in the calculation of field-free correlation and response functions. PMID- 15796701 TI - Probing transient molecular structures in photochemical processes using laser initiated time-resolved X-ray absorption spectroscopy. AB - Molecular structures during chemical processes are crucial for predicting molecular reactivity and reaction mechanisms. Using a laser pulse as an internal clock for starting fundamental chemical processes, molecular structural dynamics can be characterized by coherent vibrational motions and by incoherent transitions between different intermediate states. Recent developments in pulsed X-ray facilities allow structural determination of discrete excited states and reaction intermediates using laser-initiated time-resolved X-ray absorption spectroscopy (LITR-XAS). Moreover, femtosecond X-ray sources have begun making significant contributions in monitoring coherent molecular motions. This review summarizes recent developments in the field, including technical and scientific challenges as well as several examples involving excited state molecular structure and electronic configuration determinations. Future applications of this technique with high time resolution will enable visualization of fundamental chemical events in many systems and further our understanding in photochemistry. PMID- 15796702 TI - Vibrational autoionization in polyatomic molecules. AB - The vibrationally autoionizing Rydberg states of small polyatomic molecules provide a fascinating laboratory in which to study fundamental nonadiabatic processes. In this review, recent results on the vibrational mode dependence of vibrational autoionization are discussed. In general, autoionization rates depend strongly on the character of the normal mode driving the process and on the electronic character of the Rydberg electron. Although quantitative calculations based on multichannel quantum defect theory are available for some polyatomic molecules, including H3, only qualitative information exists for most molecules. This review shows how qualitative information, such as Walsh diagrams along different normal coordinates of the molecule, can provide insight into the vibrational autoionization rates. PMID- 15796703 TI - Detecting microdomains in intact cell membranes. AB - Current models for cellular plasma membranes focus on spatial heterogeneity and how this heterogeneity relates to cell function. In particular, putative lipid raft membrane domains have been postulated to exist based in large part on the results that a significant fraction of the membrane is detergent insoluble and that molecules facilitating key membrane processes like signal transduction are often found in the detergent-resistant membrane fraction. Yet, the in vivo existence of lipid rafts remains extremely controversial because, despite being sought for more than a decade, evidence for their presence in intact cell membranes is inconclusive. In this review, a variety of experimental techniques that have been or might be used to look for lipid microdomains in intact cell membranes are described. Experimental results are highlighted and the strengths and limitations of different techniques for microdomain identification and characterization are assessed. PMID- 15796704 TI - Ultrafast chemistry: using time-resolved vibrational spectroscopy for interrogation of structural dynamics. AB - Time-resolved infrared (IR) and Raman spectroscopy elucidates molecular structure evolution during ultrafast chemical reactions. Following vibrational marker modes in real time provides direct insight into the structural dynamics, as is evidenced in studies on intramolecular hydrogen transfer, bimolecular proton transfer, electron transfer, hydrogen bonding during solvation dynamics, bond fission in organometallic compounds and heme proteins, cis-trans isomerization in retinal proteins, and transformations in photochromic switch pairs. Femtosecond IR spectroscopy monitors the site-specific interactions in hydrogen bonds. Conversion between excited electronic states can be followed for intramolecular electron transfer by inspection of the fingerprint IR- or Raman-active vibrations in conjunction with quantum chemical calculations. Excess internal vibrational energy, generated either by optical excitation or by internal conversion from the electronic excited state to the ground state, is observable through transient frequency shifts of IR-active vibrations and through nonequilibrium populations as deduced by Raman resonances. PMID- 15796705 TI - Microfluidic tools for studying the specific binding, adsorption, and displacement of proteins at interfaces. AB - A combination of temperature and concentration gradient microfluidic devices were employed to study the mechanistic details of biomacromolecule interactions at oxide interfaces. These lab-on-a-chip techniques allowed high-throughput, multiplexed data collection using only nanoliters of analyte. The three examples presented demonstrate rapid data acquisition relative to standard methods. First, we show ligand-receptor binding data for multivalent binding between membrane bound ligands and incoming aqueous proteins with several binding pockets. A model is described for obtaining both the first and second dissociation constant for the reaction. The second example employs temperature gradient microfluidics to study the thermoresponsive properties of polymers and proteins. Both the folding mechanism and subsequent formation of an aqueous two-phase system were followed. Finally, these microfluidic techniques were combined with fluorescence microscopy and nonlinear optical spectroscopy to elucidate the mechanism of fibrinogen displacement from silica surfaces. This combination of methods enabled both direct and indirect observation of protein conformational changes. PMID- 15796706 TI - Ab initio quantum chemical and mixed quantum mechanics/molecular mechanics (QM/MM) methods for studying enzymatic catalysis. AB - We describe large scale ab initio quantum chemical and mixed quantum mechanics/molecular mechanics (QM/MM) methods for studying enzymatic reactions. First, technical aspects of the methodology are reviewed, including the hybrid density functional theory (DFT) methods that are typically employed for the QM aspect of the calculations, and various approaches to defining the interface between the QM and MM regions in QM/MM approaches. The modeling of the enzymatic catalytic cycle for three examples--methane monooxygenase, cytochrome P450, and triose phosphate isomerase--are discussed in some depth, followed by a brief summary of other systems that have been investigated by ab initio methods over the past several years. Finally, a discussion of the qualitative and quantitative conclusions concerning enzymatic catalysis that are available from modern ab initio approaches is presented, followed by a conclusion briefly summarizing future prospects. PMID- 15796707 TI - Fourier transform infrared vibrational spectroscopic imaging: integrating microscopy and molecular recognition. AB - The recent development of Fourier transform infrared (FTIR) spectroscopic imaging has enhanced our capability to examine, on a microscopic scale, the spatial distribution of vibrational spectroscopic signatures of materials spanning the physical and biomedical disciplines. Recent activity in this emerging area has concentrated on instrumentation development, theoretical analyses to provide guidelines for imaging practice, novel data processing algorithms, and the introduction of the technique to new fields. To illustrate the impact and promise of this spectroscopic imaging methodology, we present fundamental principles of the technique in the context of FTIR spectroscopy and review new applications in various venues ranging from the physical chemistry of macromolecular systems to the detection of human disease. PMID- 15796708 TI - Transport spectroscopy of chemical nanostructures: the case of metallic single walled carbon nanotubes. AB - Transport spectroscopy, a technique based on current-voltage measurements of individual nanostructures in a three-terminal transistor geometry, has emerged as a powerful new tool to investigate the electronic properties of chemically derived nanostructures. In this review, we discuss the utility of this approach using the recent studies of single-nanotube transistors as an example. Specifically, we discuss how transport measurements can be used to gain detailed insight into the electronic motion in metallic single-walled carbon nanotubes in several distinct regimes, depending on the coupling strength of the contacts to the nanotubes. Measurements of nanotube devices in these different conductance regimes have enabled a detailed analysis of the transport properties, including the experimental determination of all Hartree-Fock parameters that govern the electronic structure of metallic nanotubes and the demonstration of Fabry-Perot resonators based on the interference of electron waves. PMID- 15796709 TI - Ultrafast electron transfer at the molecule-semiconductor nanoparticle interface. AB - Electron transfer across the molecule-semiconductor interface is a fundamental process that is relevant to many applications of nanoparticles, such as dye sensitized solar cells and molecular electronics. This review summarizes recent progress in understanding electron transfer dynamics from molecular adsorbates to semiconductor nanoparticles. Photoexcitation of molecular adsorbates to their excited states is followed by electron injection into semiconductor nanoparticles. The products of electron injection (oxidized adsorbate and electrons in semiconductor) are monitored by their electronic and vibrational spectra, allowing direct measurement of injection rate. The dependence of injection rate on the properties of semiconductor nanoparticle, molecular adsorbate, intervening bridging and anchoring group, and interfacial environment are discussed and compared with Marcus theory of interfacial electron transfer. PMID- 15796710 TI - Heat capacity in proteins. AB - Heat capacity (Cp) is one of several major thermodynamic quantities commonly measured in proteins. With more than half a dozen definitions, it is the hardest of these quantities to understand in physical terms, but the richest in insight. There are many ramifications of observed Cp changes: The sign distinguishes apolar from polar solvation. It imparts a temperature (T) dependence to entropy and enthalpy that may change their signs and which of them dominate. Protein unfolding usually has a positive deltaCp, producing a maximum in stability and sometimes cold denaturation. There are two heat capacity contributions, from hydration and protein-protein interactions; which dominates in folding and binding is an open question. Theoretical work to date has dealt mostly with the hydration term and can account, at least semiquantitatively, for the major Cp related features: the positive and negative Cp of hydration for apolar and polar groups, respectively; the convergence of apolar group hydration entropy at T approximately 112 degrees C; the decrease in apolar hydration Cp with increasing T; and the T-maximum in protein stability and cold denaturation. PMID- 15796712 TI - Time-resolved spectroscopy of organic dendrimers and branched chromophores. AB - Organic dendrimers have been considered for a number of optical applications and are now of great interest for the purpose of enhanced nonlinear optical effects. In order to understand the mechanism of the enhanced effects in branched structures it is important to probe the fundamental excitations and the degree of intramolecular interactions utilizing various spectroscopic techniques. In this review, the nonlinear optical and excited state dynamics of different dendritic and other branching chromophore structures are discussed. The methods of two photon absorption, time-resolved fluorescence, transient absorption, and three pulse photon echo peak shift are discussed in regards to the degree of intramolecular coupling in the macromolecular systems. These techniques are also used for a comparison of the dynamics in the linear molecular analog systems as well. Thus, this review focuses on the aspect of intramolecular interactions in a branched system and its importance to enhanced nonlinear optical effects useful for modern optical devices. PMID- 15796711 TI - Metal to insulator transitions in clusters. AB - The strict criterion for metallicity, a finite density of states (DOS) at the Fermi energy (E(F)), cannot be applied to clusters because energy levels are always discrete in a system of finite size. We propose an alternative definition whereby clusters can be considered metallic when the gap between occupied and unoccupied states at E(F) is consistently smaller than or equal to the Kubo band gap delta. We use the experimental findings of photoelectron spectroscopy of anionic clusters to analyze band gaps of various cluster families. Monovalent clusters (alkali and noble metals) grossly follow the shell structure pattern, producing band gaps smaller than delta for most cluster sizes, with some exceptional sizes exhibiting electronic shell closure or symmetry-induced band gaps. Among the bivalent metals, only mercury shows consistent band gap closure with increasing cluster size, that is a simple insulator-metal transition. Other bivalent elements such as Zn and Mg exhibit a much more complicated behavior. We also briefly discuss complex cluster families such as aluminum and transition metals. PMID- 15796713 TI - An abortive apoptotic pathway induced by singlet oxygen is due to the suppression of caspase activation. AB - Singlet oxygen causes the cytotoxic process of tumour cells in photodynamic therapy. The mechanism by which singlet oxygen damages cells is, however, not fully understood. To address this issue, we synthesized and used two types of endoperoxides, MNPE (1-methylnaphthalene-4-propionate endoperoxide) and NDPE (naphthalene-1,4-dipropionate endoperoxide), that generate defined amounts of singlet oxygen at 37 degrees C with similar half lives. MNPE, which is more hydrophobic than NDPE, induced the release of cytochrome c from mitochondria into the cytosol and exhibited cytotoxicity, but NDPE did not. RBL cells, a rat basophil leukaemia-derived line, that overexpress phospholipid hydroperoxide glutathione peroxidase in mitochondria were found to be highly resistant to the cytotoxic effect of MNPE. MNPE treatment induced much less DNA ladder formation and nuclear fragmentation in cells than etoposide treatment, even though these treatments induced a similar extent of cellular damage. Singlet oxygen inhibited caspase 9 and 3 activities directly and also suppressed the activation of the caspase cascade. Collectively, these data suggest that singlet oxygen triggers an apoptotic pathway by releasing cytochrome c from mitochondria via the peroxidation of mitochondrial components and results in cell death that is different from typical apoptosis, because of the abortive apoptotic pathway caused by impaired caspase activation. PMID- 15796714 TI - Comparative characterization of rat deoxyribonuclease 1 (Dnase1) and murine deoxyribonuclease 1-like 3 (Dnase1l3). AB - Deoxyribonuclease 1 (DNASE1, DNase I) and deoxyribonuclease 1-like 3 (DNASE1L3, DNase gamma, DNase Y, LS-DNase) are members of a DNASE1 protein family that is defined by similar biochemical properties such as Ca2+/Mg2+-dependency and an optimal pH of about 7.0 as well as by a high similarity in their nucleic acid and amino acid sequences. In the present study we describe the recombinant expression of rat Dnase1 and murine Dnase1l3 as fusion proteins tagged by their C-terminus to green fluorescent protein in NIH-3T3 fibroblasts and bovine lens epithelial cells. Both enzymes were translocated into the rough endoplasmic reticulum, transported along the entire secretory pathway and finally secreted into the cell culture medium. No nuclear occurrence of the nucleases was detectable. However, deletion of the N-terminal signal peptide of both nucleases resulted in a cytoplasmic and nuclear distribution of both fusion proteins. Dnase1 preferentially hydrolysed 'naked' plasmid DNA, whereas Dnase1l3 cleaved nuclear DNA with high activity. Dnase1l3 was able to cleave chromatin in an internucleosomal manner without proteolytic help. By contrast, Dnase1 was only able to achieve this cleavage pattern in the presence of proteases that hydrolysed chromatin-bound proteins. Detailed analysis of murine sera derived from Dnase1 knockout mice revealed that serum contains, besides the major serum nuclease Dnase1, an additional Dnase1l3-like nucleolytic activity, which, in co operation with Dnase1, might help to suppress anti-DNA autoimmunity by degrading nuclear chromatin released from dying cells. PMID- 15796715 TI - A novel assay method for an amino acid racemase reaction based on circular dichroism. AB - We have established a novel assay method based on circular dichroism that can be used for the kinetic study of the activity of amino acid racemases, such as ALR (alanine racemase). Although an enzyme-coupled assay method has been used to measure racemase activity, the CD method is superior to the enzyme assay because it can accurately determine the immediate changes of an enantiomer on racemization between its L- and D-forms. The enzyme-coupled assay requires D amino acid oxidase, which is inactivated by an inhibitor of ALR, D-cycloserine. This indicates that the inhibitory kinetic study for ALR with D-cycloserine by the enzyme-coupled assay method is restricted to the analysis of only the reaction resulting in the formation of L-Ala from D-Ala. However, since the CD assay does not require the coupled enzyme, it can be used to comprehensively evaluate the reactions that result in the formation both of D-Ala from L-Ala and of L-Ala from D-Ala at several substrate concentrations. Streptomyces ALR also catalyses the formation of D-Ser from L-Ser and of L-Ser from D-Ser, but the catalytic constants (kcat) are 4- and 10-fold lower than those for the formation of D-Ala from L-Ala and of L-Ala from D-Ala respectively. PMID- 15796716 TI - Functional and structural characterization of the myoglobin from the polychaete Ophelia bicornis. AB - The myoglobin of the polychaete annelid Ophelia bicornis was isolated, purified to homogeneity and characterized. The primary structure, obtained from cDNA and protein sequencing, consists of 139 amino acid residues. The alignment with other globin sequences showed that O. bicornis myoglobin misses the pre-A helix and the first six residues of the A helix. The presence of a PheB10-GlnE7 haem distal residue pair is in agreement with the measured oxygen affinity (P50=0.85 mmHg; 1 mmHg=0.133 kPa) and the only slightly higher autoxidation rate constant (0.28 h( 1)) with respect to that of the sperm whale myoglobin mutant E7 His-->Gln (0.21 h(-1)) and to elephant myoglobin (0.1 h(-1)). Oxygen-binding co-operativity was found to be absent under all the examined experimental conditions. The resistance of O. bicornis myoglobin towards autoxidation seems to confirm the important role of part of the A helix in the stability of the globin. The higher pKa of the acid alkaline ferric transition of O. bicornis with respect to Asian elephant myoglobin, as well as the higher absorbance ratio of its ferric form to the oxy form measured in the Soret region (gammamet/gammaoxy) with respect to that of the African elephant myoglobin, suggested a stronger interaction between the distal glutamine and the water molecule at the sixth co-ordinate position. PMID- 15796719 TI - 15th European Congress of Clinical Microbiology and Infectious Diseases, Copenhagen, Denmark, 2-5 April 2005. Abstracts. PMID- 15796717 TI - Recruitment of MKLP1 to the spindle midzone/midbody by INCENP is essential for midbody formation and completion of cytokinesis in human cells. AB - The INCENP (inner centromere protein) is a chromosomal passenger protein that plays multiple roles in regulating mitosis and cytokinesis. The MKLP1 (mitotic kinesin-like protein) is a component of centralspindlin complex that has been implicated in assembly of midzone/midbody during mitosis and is essential for cytokinesis. In the present study, we investigated functions of INCNEP and MKLP1 and their interplay in regulating spindle midzone/midbody formation and cytokinesis in human cells. Immunofluorescence and live-cell imaging analyses have shown that, in addition to multiple chromosome segregation defects, cells that lacked INCENP by RNAi (RNA interference) exhibit abnormal spindle midzone/midbody formation, resulting in formation of binucleated/multinucleated cells. Suppression of MKLP1 expression by siRNA (small interfering RNA) did not cause any abnormality of chromosome segregation and midzone formation, but abrogated midbody formation and completion of cytokinesis. Furthermore, we show that INCENP is required for recruiting MKLP1 to the spindle midzone/midbody. Three-dimensional reconstruction imaging analysis suggests that recruitment of MKLP1 to the midzone/midbody by INCENP is a crucial step for the midbody formation and completion of cytokinesis in mammalian cells. PMID- 15796718 TI - Characterization of a novel NADP(+)-dependent D-arabitol dehydrogenase from the plant pathogen Uromyces fabae. AB - We have identified and characterized a novel NADP(+)-dependent D-arabitol dehydrogenase and the corresponding gene from the rust fungus Uromyces fabae, a biotrophic plant pathogen on broad bean (Vicia faba). The new enzyme was termed ARD1p (D-arabitol dehydrogenase 1). It recognizes D-arabitol and mannitol as substrates in the forward reaction, and D-xylulose, D-ribulose and D-fructose as substrates in the reverse reaction. Co-factor specificity was restricted to NADP(H). Kinetic data for the major substrates and co-factors are presented. A detailed analysis of the organization and expression pattern of the ARD1 gene are also given. Immunocytological data indicate a localization of the gene product predominantly in haustoria, the feeding structures of these fungi. Analyses of metabolite levels during pathogenesis indicate that the D-arabitol concentration rises dramatically as infection progresses, and D-arabitol was shown in an in vitro system to be capable of quenching reactive oxygen species involved in host plant defence reactions. ARD1p may therefore play an important role in carbohydrate metabolism and in establishing and/or maintaining the biotrophic interaction in U. fabae. PMID- 15796721 TI - Chronic heart failure. PMID- 15796722 TI - Reporting clinical research: an example of balancing a simple clinical message with data required to help future research. PMID- 15796723 TI - Victorian Emergency Minimum Dataset: factors that impact upon the data quality. AB - OBJECTIVE: The Victorian Emergency Minimum Dataset (VEMD) records details of approximately 80% of Victoria's ED presentations. Its usefulness for quality assurance and research relies on the data being both complete and accurate. We aimed to determine the factors that impact adversely on the collection of high quality VEMD data. METHODS: The study was a voluntary, anonymous, cross-sectional survey of a range of ED staff (medical, nursing, clerical) who collect and enter data into the VEMD. Nine of the 28 hospitals that contribute to the VEMD were surveyed. The questionnaire was purpose-designed and self-administered. RESULTS: A total of 218 staff participated (response rate 95%). Six different software types were used, with 40% of respondents using the Pickware (MCAT) system. There was no consistency of ED personnel for the completion of specific data fields. One hundred and twenty-six (56%) respondents had heard of the VEMD, 67 (29%) had had its structure and purpose explained and 65 (30%) had been trained to enter data. Ninety-seven (45%) respondents knew what the VEMD data was used for, 38 (17%) knew they could request VEMD data for their own use and 17 (7.8%) had done so. Time constraints, software problems and lack of formal orientation and training in data entry were reported as the most important factors impacting adversely upon quality data entry. CONCLUSION: Staff knowledge of the VEMD system and its uses are poor. Numerous factors impact on the quality of data entered and interventions aimed at improving staff education, training and feedback and software are indicated. PMID- 15796724 TI - How accurate is weight estimation in the emergency department? AB - OBJECTIVE: To determine the accuracy of medical staff, nursing staff and patients for estimating weight in an ED population. METHODS: This is a prospective, observational study. Medical staff, nursing staff and patients were asked to estimate patient weight that was then measured. The main outcome was average per cent error in weight estimation for each group. RESULTS: Average per cent error in estimates was 3.9% for patients (95% CI 3.6-4.1%), 7.7% (95% CI 7.2-8.2%) for nurses and 11% (95% CI 10.2-11.7%) for physicians. Ninety-one per cent of patients (95% CI 90-93%), 78% of nurses (95% CI 75-80%) and 59% of physicians (95% CI 56-63%) made weight estimates accurate to within 10% of actual weight. CONCLUSION: Patients are generally accurate in estimating their true weight and health care workers showed only moderate accuracy. Where possible, drug dose calculations should be based on measured weight and if this is not possible, patient estimate of weight should be sought. Health care worker estimation should be used only when this is not possible. PMID- 15796725 TI - Patient perceptions of privacy infringements in an emergency department. AB - OBJECTIVE: To identify the nature, severity, impact, frequency and risk factors for patient perceived privacy infringements in the ED of St Vincent's Health Melbourne with 32,000 emergency attendances per annum. METHODS: Patients 18 years and older attending emergency over a 2-week period were offered a nine-item questionnaire using a Likert scale. A privacy incident was defined as: (i) overhearing medical or personal information; (ii) being overheard; (iii) having private body parts exposed or (iv) seeing others' body parts. Differences between demographic, emergency environment, length of stay and other factors suspected of affecting patient privacy were quantified. RESULTS: From 1169 emergency presentations, 235 patients returned questionnaires, with 105 of these (45%) reporting a total of 159 privacy incidents. Seventy-eight patients (33%, 95% CI 28-36%) reported a definite privacy incident and 81 (35%, 95% CI 29-37%) reported a probable privacy incident. Ninety-six patients (41%, 95% CI 35-47%) reported overhearing other patient's conversations with the staff whereas 36 (15%, 95% CI 11-21%) felt their conversations with staff were overheard by others. Twenty seven patients (11%, 95% CI 6-14%) experienced or observed inappropriate exposure of private body parts. Ten patients (4%, 95% CI 2-7%) changed or withheld information from staff because they felt others may overhear it and two refused part of their physical examination because they felt they may be seen by inappropriate people. Twenty-four patients (10%, 95% CI 6-14%) did not have their expectations of privacy met. The longer a patient was in emergency the greater the number of privacy incidents they experienced (Spearman correlation P < 0.01). Patients in walled cubicles experienced fewer privacy incidents than those in curtained cubicles (P < 0.05). CONCLUSIONS: Patient privacy incidents occur frequently in an ED, risk factors being length of stay and absence of a walled cubicle. Patients who have their conversations overheard are more likely to withhold information from staff and less likely to have had their expectations of privacy met. PMID- 15796726 TI - Challenges in arranging interhospital transfers from a small regional hospital: an observational study. AB - OBJECTIVE: To describe the process of interhospital transfer from a small regional hospital ED. METHODS: Prospective observational case series of patients transferred from the ED of West Gippsland Hospital (WGH), Victoria from February to August 2003. Outcome measures were expressed as the number of phone calls required to facilitate interhospital transfer, the time taken for a proposed transfer to be accepted and the total time a transferred patient spent in the ED. RESULTS: Eighty-one of 129 transferred patients had all data available. Those with missing data had similar ED length of stays, and were of similar age and acuity to those with complete data available. Mean (95% CI) values for the study population were number of phone calls: 4.7 (95% CI 3.96-5.43); time to transfer acceptance: 56.7 min (95% CI 19.1-94.8 min); and total time in ED: 307 min (95% CI 32.9-372.6 min). Men (P = 0.01), older patients (P = 0.02) and those for whom a centralized referral service were consulted (P = 0.006) were more likely to require four or more phone calls to facilitate transfer. The time until a transfer was accepted was more likely to be longer than an hour for patients who were transferred to critical care areas (P = 0.02) and those for whom a centralized referral service was consulted (P = 0.002). Patients referred to another ED spent less time in WGH ED than other transferred patients. CONCLUSION: There is wide variation in the time taken and the number of phone calls to arrange an interhospital transfer, with some patient groups requiring more effort to organize a transfer. PMID- 15796727 TI - Local anaesthetic infiltration increases the accuracy of assessment of ulnar collateral ligament injuries. AB - OBJECTIVE: To assess the effectiveness of a systematic examination of the ulnar collateral ligament (UCL) of the thumb metacarpophalangeal joint (MCPJ) prior to and post infiltration of local anaesthetic. METHODS: During the study period from 24 October 2001 to 22 February 2002, 51 patients with clinical signs suggestive of UCL injuries were identified prospectively from initial ED attendances and attendances at a subsequent review clinic. Patients were formally assessed a mean of 6.6 days post injury. A single ED Senior House Officer carried out examination before and after direct infiltration of local anaesthetic around the site of injury. Stress radiography was also performed as the 'gold standard' diagnostic test of UCL instability. RESULTS: Forty-seven patients were enrolled in the study. When reviewed by the single observer, examination prior to and post local anaesthetic infiltration revealed a degree of joint stability in 28% (95% CI 16 43%) and 98% (95% CI 88-100%) cases, respectively, compared to the gold standard. Post infiltration, this technique had a specificity of 100% (95% CI 94-100%) and a sensitivity of 87.5% (95% CI 74-95%) (P < 0.001). Stress radiography offered additional information in one patient. A total of eight patients previously underdiagnosed in the ED were found to have unstable thumb MCPJs. CONCLUSION: This simple technique improves assessment of suspected UCL injuries approximately 1 week post injury. Further studies are indicated to determine its effectiveness in the ED immediately post injury. PMID- 15796728 TI - Trauma bypass in south-eastern Sydney: an 8-year review. AB - OBJECTIVE: The New South Wales (NSW) Health Department and the Ambulance Service of NSW introduced a trauma bypass system in Sydney on 29 March 1992. This study aims to review the outcomes of trauma bypass patients brought to St George Hospital, a major trauma service in south-eastern Sydney, and to assess the performance of the current prehospital trauma triage protocol. METHODS: The St George Hospital Department of Trauma Services prospectively collected data on all trauma bypass patients for the 8-year period from 29 March 1992 to 29 March 2000. RESULTS: A total of 1990 patients were brought to hospital on trauma bypass. The average age was 32 years, 70% were men and 66% were from road traffic accidents. The positive predictive value of the prehospital triage tool for serious injury (Injury Severity Score [ISS] > 15) was 18.6% (95% CI 16.9-20.4). This is well below the benchmark previously established by the NSW Health Department Trauma System Advisory Committee. For all trauma bypass patients, 33.8% (95% CI 31.7 35.9) were discharged home from the ED. The overall death rate was 2.5% (95% CI 1.9-3.3). CONCLUSIONS: According to the proposed benchmark, current prehospital trauma triage guidelines are underperforming. This suggests that a review of the benchmarks of current local trauma systems and of the trauma triage tool is required. PMID- 15796729 TI - Management of chronic heart failure: an old disease with a new face. AB - Heart failure is a leading cause of morbidity and mortality. It affects over 5 million patients annually. There are an estimated 400,000-700,000 new cases diagnosed each year. The management of heart failure has changed significantly over the last decade. This review focuses on the pharmacologic management of systolic heart failure, and provides current recommendations for the practicing acute care provider. PMID- 15796730 TI - A double-blind, randomized trial of intravenous versus intramuscular antivenom for red-back spider envenoming. AB - OBJECTIVE: To compare the efficacy of intravenous versus intramuscular antivenom (AV) in the treatment of Red-back spider (RBS) envenoming. METHODS: Randomized, double-dummy, double-blind, multicentre trial of patients with red-back spider envenoming requiring AV treatment recruited from five hospital EDs in Western Australia. RESULTS: Thirty-five patients were recruited; two were excluded; 33 were available for initial analysis, but two who were unblinded after one ampoule of trial AV and given i.v. AV had limited data; 31 remained in the study and had more complete data. After AV, pain scores for both i.m. and i.v. groups improved rapidly. At 24 h, the i.v. group was better with a 55% absolute difference (76% vs. 21%; 95% CI 25-85% difference) in the proportion pain-free. There were no safety issues. CONCLUSIONS: Red-back spider antivenom was initially effective by both i.m. and i.v. routes. The study generates the hypothesis that at 24 h, significantly more patients are pain-free with i.v. administration. Definitive recommendations on the optimal route of administration of RBS AV await the results of further studies. PMID- 15796731 TI - Boarder belly: splenic injuries resulting from ski and snowboarding accidents. AB - OBJECTIVE: Snowboarding has increased in popularity worldwide, with an associated increase in injuries suffered by its participants with a significant proportion of these injuries being severe. We sought to understand the risk of sustaining a splenic injury in snowboarders as compared to skiers, and whether there are noteworthy differences in their characteristics at hospital admission. METHODS: A 10-year retrospective review was conducted on patients with splenic injury resulting from snowboarding or skiing, who were admitted to the principle ED and referral hospital servicing several busy downhill skiing areas. Population-based injury rates were calculated for our catchment area, using data provided by the Canadian Ski Council. RESULTS: Controlling for gender, snowboarders were six times more likely to sustain a splenic injury than skiers (P < 0.0001). The risk of splenic injury was 21.7 times greater for male snowboarders than for female snowboarders (P = 0.002). By contrast, no gender differences were observed for skiers. Snowboarders admitted to hospital with a splenic injury were significantly younger, more likely to present with an isolated injury and to required a shorter hospital stay, as compared to skiers. CONCLUSION: The risk of sustaining an injury of the spleen resulting from blunt abdominal trauma while snowboarding is significantly greater than the risk while downhill skiing. Male snowboarders have a significantly higher risk of splenic injury than female snowboarders. In the majority of cases, snowboarders sustained their injuries as a result of falls or jumps. PMID- 15796732 TI - Serious parasport injuries in Auckland, New Zealand. AB - OBJECTIVES: Parasports' comprise parachuting, skydiving, paragliding, parapenting, parascending, parasailing and hang-gliding, and may involve rapid deceleration on landing. The aims of this study were to determine the spectrum of parasport-related trauma presenting to Auckland City Hospital and to make recommendations to reduce further injury. METHODS: The Auckland City Hospital Trauma Registry was used to identify all patients requiring admission with serious injury as a result of parasport incidents over an 8-year period. Demographic and injury-related data were then analysed. RESULTS: Parachuting/skydiving was responsible for 66% of all serious parasport injuries and 96% of patients were men. Fractures of the lower limbs and lumbar spine were the predominant injuries. Misjudgement of landing speed and attitude resulting in injurious landing forces were the usual causes of injury. CONCLUSIONS: The magnitude of serious parasport-related trauma is greater than that indicated by previous studies in New Zealand. Appropriate use of protective clothing and close attention to safe flight planning and landing techniques may reduce the injury rate without degrading the experience of flight. PMID- 15796733 TI - The status of emergency medicine in Makira Ulawa Province, Solomon Islands. AB - This Perspective reports on the challenges that face a solo doctor in a provincial hospital in the Solomon Islands following the civil disturbances of 1998-2003. The Health Service is seriously constrained by a paucity of funding, supplies and personnel. In spite of that, a rudimentary service can be provided and lives can be saved using simple techniques and basic resources. Further training of nurses, midwives and doctors is required. Emergency medicine, as a generalist discipline, provides a foundation to improve the delivery of care to the acutely ill and injured in these circumstances. PMID- 15796734 TI - A mysterious case of chest pain. AB - Atrial myxomas can present a diagnostic dilemma attributed to their broad spectrum of clinical presentations. We report the case of a 46-year-old woman in whom the diagnosis of atrial myxoma initially eluded cardiologists despite the large size of her tumour. PMID- 15796735 TI - Death by oral ingestion of iodine. AB - We report the clinical and toxicological features of a case of fatal iodine ingestion, and summarize the physiology, clinical characteristics and management of iodine ingestion. Physicians are likely to have little experience in managing such patients, particularly with the advent of less toxic preparations, and therefore need ready access to management guidelines. This case also highlights the potential for substances to retain their toxicity over long periods of time. PMID- 15796737 TI - Drotrecogin alfa: too early to give it early. PMID- 15796738 TI - Intralipid ameliorates thiopentone induced respiratory depression in rats: investigative pilot study. PMID- 15796739 TI - Big black spider bite from the desert region of Western Australia. PMID- 15796744 TI - Detection and analysis of alternative splicing in the silkworm by aligning expressed sequence tags with the genomic sequence. AB - We identified 277 alternative splice forms in silkworm genes based on aligning expressed sequence tags with genomic sequences, using a transcript assembly program. A large fraction (74%) of these alternative splices are located in protein-coding regions and alter protein products, whereas only 26% are in untranslated regions. From the alternative splices located in protein-coding regions, some (43%) affect protein domains that bind various biological molecules. The vast majority of the detected alternative forms in this study appear to be novel, and potentially affect biologically meaningful control of function in silkworm genes. Our results indicate that alternative splicing in silkworm largely produces protein diversity and functional diversity, and is a widely used mechanism for regulating gene expression. PMID- 15796745 TI - Midgut and salivary gland transcriptomes of the arbovirus vector Culicoides sonorensis (Diptera: Ceratopogonidae). AB - Numerous Culicoides spp. are important vectors of livestock or human disease pathogens. Transcriptome information from midguts and salivary glands of adult female Culicoides sonorensis provides new insight into vector biology. Of 1719 expressed sequence tags (ESTs) from adult serum-fed female midguts harvested within 5 h of feeding, twenty-eight clusters of serine proteases were derived. Four clusters encode putative iron binding proteins (FER1, FERL, PXDL1, PXDL2), and two clusters encode metalloendopeptidases (MDP6C, MDP6D) that probably function in bloodmeal catabolism. In addition, a diverse variety of housekeeping cDNAs were identified. Selected midgut protease transcripts were analysed by quantitative real-time PCR (q-PCR): TRY1_115 and MDP6C mRNAs were induced in adult female midguts upon feeding, whereas TRY1_156 and CHYM1 were abundant in midguts both before and immediately after feeding. Of 708 salivary gland ESTs analysed, clusters representing two new classes of protein families were identified: a new class of D7 proteins and a new class of Kunitz-type protease inhibitors. Additional cDNAs representing putative immunomodulatory proteins were also identified: 5' nucleotidases, antigen 5-related proteins, a hyaluronidase, a platelet-activating factor acetylhydrolase, mucins and several immune response cDNAs. Analysis by q-PCR showed that all D7 and Kunitz domain transcripts tested were highly enriched in female heads compared with other tissues and were generally absent from males. The mRNAs of two additional protease inhibitors, TFPI1 and TFPI2, were detected in salivary glands of paraffin-embedded females by in situ hybridization. PMID- 15796746 TI - Expressed sequence tags from Diabrotica virgifera virgifera midgut identify a coleopteran cadherin and a diversity of cathepsins. AB - The Western corn rootworm is the major pest of corn in the USA and has recently become the target for insect-resistant transgenic crops. Transgenic crops have switched the focus for identifying insecticide targets from the insect nervous system to the midgut. Here we describe a collection of 691 sequences from the Western corn rootworm midgut, 27% of which predict proteins with no matches in current databases. Of the remaining sequences, most predict proteins with either catalytic (62%) or binding (19%) functions, as expected for proteins expressed in the insect midgut. The utility of this approach for the identification of targets for novel toxins is demonstrated by analysis of the first coleopteran cadherin gene, a putative Bt receptor, and a large class of cysteine-proteases, the cathepsins. PMID- 15796747 TI - Genetic variation in the sand fly salivary protein, SP-15, a potential vaccine candidate against Leishmania major. AB - SP-15 is a sandfly salivary protein that provides strong protection against cutaneous leishmaniasis, caused by Leishmania major, and has been proposed as a potential vaccine against this disease. To investigate possible antigenic variation in this protein, we examined genetic polymorphism of SP-15 in 100 Phlebotomus papatasi sandflies, from a natural population from Sudan and four laboratory colonies from Egypt, Jordan, Israel and Saudi Arabia. We found that although many variants of SP-15 may be found in nature, differences among them are minimal (mean+/-SD pairwise differences=1.69+/-0.83% for forty nucleotide sequences and 3.06+/-1.13% for thirty amino acid sequence variants). Analysis of proportions of synonymous and non-synonymous substitutions indicated that SP-15 is not under diversifying selection. Our results suggest that a vaccine based on SP-15 protein should result in a uniform immune response. PMID- 15796748 TI - Cell cycle profiles of EcR, USP, HR3 and B cyclin mRNAs associated to 20E-induced G2 arrest of Plodia interpunctella imaginal wing cells. AB - Using the IAL-PID2 cell line established from pupally committed imaginal wing discs of Plodia interpunctella, we have investigated the dynamics of cellular and molecular events involved in the G2/M arrest. We have first cloned a cDNA sequence named PIUSP-2 that likely encodes a homologue of the Ultraspiracle-2 isoform of Manduca sexta. When the IAL-PID2 cells were exposed to a 8 h 20E treatment applied at different times of the cell cycle, an optimal period of sensitivity of cells to 20E, in inducing G2 arrest, was determined at the S/G2 transition. Using cDNA probes specifically designed from Plodia B cyclin (PcycB), ecdysone receptor B1-isoform (PIEcR-B1) and HR3 transcription factor (PHR3), we provide evidence that the 20E-induced G2 arrest was correlated to a high induction of PHR3, PIEcR-B1, PIUSP-2 mRNAs at the S/G2 transition and a decrease in PcycB mRNA level at the end of G2 phase. PMID- 15796749 TI - cpbAg1 encodes an active carboxypeptidase B expressed in the midgut of Anopheles gambiae. AB - We previously used differential display to identify several Anopheles gambiae genes, whose expression in the mosquito midgut was regulated upon ingestion of Plasmodium falciparum. Here, we report the characterization of one of these genes, cpbAg1, which codes for the first zinc-carboxypeptidase B identified in An. gambiae and in any insect. Expression of cpbAg1 in baculovirus gave rise to an active enzyme, and determination of the N-terminal amino acids confirmed that CPBAg1 contains a signal peptide and a pro-peptide, typical features of digestive zinc carboxypeptidases. cpbAg1 mRNA was mainly produced in the mosquito midgut, where it accumulated in unfed females and was rapidly down-regulated upon blood feeding. Annotation of the An. gambiae genome predicts twenty-three sequences coding for zinc-carboxypeptidases of which only two (cpbAg1 and cpbAg2) are expressed at a significant level in the mosquito midgut. PMID- 15796750 TI - A role for alcohol dehydrogenase in the Drosophila immune response? AB - In a recent study Drosophila larvae were injected with bacterial lipopolysaccharide (LPS) suspended in 1% ethanol and differentially induced protein fractions were identified. The levels of several proteins, including alcohol dehydrogenase (ADH), increased in LPS-treated flies and were labelled as immune response proteins. However, because control larvae were not injected with ethanol alone the identified proteins could represent a response to ethanol. Here, we injected Drosophila larvae with combinations of ethanol and LPS. While ADH activity increased in larvae receiving 1% ethanol, it was not increased after LPS injection. These results suggest that ADH plays no role in the Drosophila immune response, and that other proteins identified in the previous study may instead mediate ethanol tolerance in flies and other organisms. PMID- 15796751 TI - Independent mutations in the Rdl locus confer dieldrin resistance to Anopheles gambiae and An. arabiensis. AB - Substitutions of a conserved alanine residue in the Rdl locus coding for a gamma aminobutyric acid (GABA) receptor subunit with serine or glycine confer resistance to dieldrin in various insect species. Here, we show that alanine to glycine substitution in the Rdl locus of the malaria vector, Anopheles gambiae, is genetically linked to resistance to dieldrin. An alanine to serine substitution developed independently in a dieldrin resistant strain of An. arabiensis. An allele-specific polymerase chain reaction (PCR) assay was able to differentiate dieldrin resistant and susceptible mosquitoes. PMID- 15796752 TI - Identification and molecular characterization of a prophenoloxidase involved in Aedes aegypti chorion melanization. AB - Chorion melanization is a vital biochemical event for the survival of mosquito eggs in the environment. This study describes the identification and molecular characterization of a prophenoloxidase (proPO) involved in chorion melanization in Aedes aegypti by various biochemical and molecular techniques. Results revealed that transcription of the chorion proPO occurs only in adults, blood feeding greatly stimulated its transcription and haemocytes are responsible for its transcription. Our study provides a solid basis for suggesting an essential role of the isolated proPO in chorion melanization during chorion hardening and also raises fundamental questions regarding its transportation and distribution in the chorion. This study should serve as a useful reference towards functional elucidation of other proPOs in mosquitoes. PMID- 15796754 TI - An Anopheles gambiae salivary gland promoter analysis in Drosophila melanogaster and Anopheles stephensi. AB - Regulatory regions driving gene expression in specific target organs of the African malaria vector Anopheles gambiae are of critical relevance for studies on Plasmodium-Anopheles interactions as well as to devise strategies for blocking malaria parasite development in the mosquito. In order to identify an appropriate salivary gland promoter we analysed the transactivation properties of genomic fragments located just upstream of the An. gambiae female salivary gland-specific genes AgApy and D7r4. An 800 bp fragment from the AgApy gene directed specific expression of the LacZ reporter gene in the salivary glands of transgenic Anopheles stephensi. However, expression levels were lower than expected and the transgene was expressed in the proximal-rather than in the distal-lateral lobes of female glands. Surprisingly, a promoter fragment from the D7r4 gene conferred strong tissue-specific expression in Drosophila melanogaster but only low transcription levels in transgenic An. stephensi. These results imply a certain conservation of gland-specific control elements between the fruit fly and the mosquito suggesting that an increased degree of complexity, probably connected to the evolution of haematophagy, underlies the regulation of tissue-specific expression in mosquito female salivary glands. PMID- 15796753 TI - A novel fatty acid binding protein produced by teratocytes of the aphid parasitoid Aphidius ervi. AB - Aphidius ervi is an endophagous braconid, parasitoid of the pea aphid, Acyrthosiphon pisum. A. ervi teratocytes, deriving from the dissociation of the embryonic serosa, synthesize and release two major proteins into the host haemocoel. The gene of one of these proteins has been cloned and characterized. This gene codes for a 15.8 kDa protein belonging to the fatty acid binding protein (FABP) family, named Ae-FABP (A. ervi-FABP). It is abundantly present in the host haemolymph when the parasitoid larva attains its maximum growth rate. The recombinant Ae-FABP binds to fatty acids in vitro, showing a high affinity to C14-C18 saturated fatty acids and to oleic and arachidonic acid. The possible nutritional role for the parasitoid larva of Ae-FABP is discussed. PMID- 15796755 TI - Genome size and organization in the blacklegged tick, Ixodes scapularis and the Southern cattle tick, Boophilus microplus. AB - Genome sizes and the organization of repetitive DNA were determined in the hard ticks Ixodes scapularis and Boophilus microplus using reassociation kinetics. The I. scapularis genome contains approximately 2.15 pg (2.1x10(3) Mbp) of DNA and consists of no foldback (FB), 27% highly repetitive (HR), 39% moderately repetitive (MR), and 34% unique DNA. The B. microplus genome contains 7.5 pg (7.1x10(3) Mbp) DNA, and consists of 0.82% FB, 31% HR, 38% MR, and 30% unique DNA. In both species, repetitive sequences occur in a mixture of long and short period interspersion but most (65-80%) of the DNA follows a pattern of short period interspersion. Genome size and organization in the three tick species so far examined are distinct from other arthropods in having a greater proportion of MR, a lower proportion of unique and HR DNA of very low sequence complexity. PMID- 15796756 TI - Inhibition by lipopolysaccharide of naloxone-induced luteinising hormone secretion is accompanied by increases in corticotropin-releasing factor immunoreactivity in hypothalamic paraventricular neurones in female rats. AB - We have recently reported that lipopolysaccharide (LPS), a bacterial endotoxin, inhibits steroid-induced as well as naloxone-induced luteinising hormone (LH) secretion in ovariectomised oestrogen-primed rats. In the present study, we examined whether corticotropin-releasing factor (CRF) may be involved in the LPS induced inhibition of LH secretion. Unanaesthetised rats were treated with an intravenous (i.v.) injection of LPS (10 microg) or saline, followed by an i.v. injection of naloxone (20 mg/kg). After sequential blood samples were collected for determination of serum LH concentrations, the brains were fixed and CRF immunoreactivity was examined histochemically. In control rats receiving saline injections, only a small number of CRF-immunoreactive (ir) cells were found in the parvocellular portion of the hypothalamic paraventricular nucleus (PVN), and naloxone significantly increased serum LH concentrations within 10 min. By contrast, in LPS-treated rats, the number of CRF-ir cells was significantly greater than that in control rats, and the effect of naloxone was completely abolished. In a separate experiment, an intracerebroventricular injection of 5 microg CRF inhibited naloxone-induced LH release, mimicking the effect of LPS. These results suggest that LPS stimulates production of CRF in PVN neurones, which in turn inhibits LH secretion without opioidergic mediation. PMID- 15796757 TI - Nitration and increased alpha-synuclein expression associated with dopaminergic neurodegeneration in equine pituitary pars intermedia dysfunction. AB - Equine pituitary pars intermedia dysfunction (PPID) is a spontaneously occurring progressive disease affecting aged horses and ponies. The pathogenesis of PPID is poorly understood, but the available evidence supports a loss of dopaminergic inhibition of the melanotropes of the pars intermedia. Horses with PPID have increased plasma concentrations of pars intermedia pro-opiomelanocortin-derived peptides that decrease in response to dopamine or dopamine agonist administration. Dopamine and dopamine metabolite concentrations are decreased in the pars intermedia of affected horses compared to age-matched control horses. Horses with disease that are treated with the dopamine agonist pergolide show improvement in clinical signs and normalisation of diagnostic test results. In the present study, immunohistochemical evaluation of pituitary and hypothalamic tissue demonstrated reduced tyrosine hydroxylase immunoreactivity in affected horses compared to age-matched and young controls, supporting the role of dopaminergic neurodegeneration in PPID. In addition, immunohistochemical evaluation revealed an increase in the oxidative stress marker, 3-nitrotyrosine and in nerve terminal protein, alpha-synuclein that colocalised in the pars intermedia of horses with disease. These findings suggest a role for nitration of overexpressed alpha-synuclein in the pathogenesis of neurodegeneration in PPID. PMID- 15796758 TI - Male and female prolactin receptor mRNA expression in the brain of a biparental and a uniparental hamster, phodopus, before and after the birth of a litter. AB - Prolactin receptor (PRL-R) mRNA transcript level was quantified in the choroid plexus (ChP) of a naturally biparental hamster, Phodopus campbelli, and its otherwise similar, yet nonpaternal, sibling species, Phodopus sungorus. Pair housed males and females on the day before the birth of their first litter (G17), the day after birth (L1), lactation day 5 (L5), and unpaired animals that were sexually naive, were tested. PRL-R mRNA transcript level relative to total RNA, was evaluated by reverse transcriptase-polymerase chain reaction using primers common to the long- and short-form of the PRL-R in Phodopus. In the ChP, a region implicated in prolactin transport into the central nervous system, females had the expected increase in PRL-R mRNA transcript from dioestrus to L5, consistent with known actions of prolactin. As predicted, males and females of the biparental species were similar, although PRL-R mRNA in naive males was higher than in dioestrus females. Males of the two species also differed as predicted. PRL-R mRNA transcript levels were higher in the biparental males. In addition, P. campbelli males had low PRL-R mRNA at G17 compared to L5. By contrast, non paternal P. sungorus males had elevated PRL-R mRNA transcript levels on G17 relative to unpaired males. We conclude that PRL-R mRNA in the ChP is differentially regulated before and after birth in a paternal and a nonpaternal male. PMID- 15796759 TI - Cyclic AMP-protein kinase a and protein kinase C mediate in vitro T activation of brain tyrosine hydroxylase in the female catfish Heteropneustes fossilis. AB - The present in vitro study demonstrates an involvement of both cAMP-dependent protein kinase A (PKA) and protein kinase C (PKC) signal transduction mechanisms in the triiodothyronone (T(3))-activation of forebrain (telencephalon and hypothalamus) tyrosine hydroxylase (TH) activity in the female catfish Heteropneustes fossilis. Incubations of the enzyme preparations with different concentrations of T(3) (0.15-2.4 ng/ml) stimulated TH activity over the concentrations. Similarly, coincubations of the enzyme preparations with T(3) and cAMP (1.0 mM) or cAMP-elevating drugs such as 1-methyl-3-isobutylxanthine (1.5 mM) or theophylline (1.5 mM) increased TH activity significantly over that of T(3). The stimulatory effect of TH activity with T(3) or cAMP was coincident with a low apparent K(m) and high V(max) for the cofactor, suggesting a higher affinity of the enzyme. Incubation of the enzyme preparations with PKA (H-89) and PKC (calphostin-C) inhibitors decreased basal enzyme activity significantly, with the inhibition being greater in the former group. The incubations of the enzyme preparations with T(3) or T(3) + cAMP, followed by the different inhibitors, also decreased enzyme activity. Although T(3) could not reverse the inhibitory effect of H-89, it could over-ride the effect of calphostin-C to some extent. The suppressive effect of the inhibitors could be related to a high apparent K(m) and low V(max) for the cofactor. The evidence strongly suggests a nongenomic action of T(3) on TH activity via the cell signalling pathways, for which the cAMP dependent PKA appears to be the major regulatory mechanism. PMID- 15796760 TI - Oestrogen requires the insulin-like growth factor-I receptor for stimulation of prolactin synthesis via mitogen-activated protein kinase. AB - Sex steroids and growth factors interact at the intracellular level in a variety of tissues to control numerous physiological functions. Oestrogen is known to stimulate prolactin synthesis and secretion, but the effect of insulin-like growth factor (IGF)-I is less clear. We used GH3 cells, a somatolactotroph cell line, to study the interaction of 17beta-oestradiol (E(2)) and IGF-I on prolactin protein levels and the intracellular mechanisms involved. Cell cultures were treated with E(2) (10 nM) and/or IGF-I (10 ng/ml) for 8 h. The real-time reverse transcriptase-polymerase chain reaction, Western blot and enzyme-immunoassay were used to determine changes in prolactin mRNA and protein levels. At this time point, there were no significant changes in cell number, prolactin mRNA expression, or the amount of secreted prolactin. However, E(2) increased intracellular prolactin concentrations. IGF-I alone had no effect, but blocked the stimulatory effect of E(2). MAPK (ERK1/2) activation, as determined by Western blot analysis, increased with both E(2) and IGF-I, but not with the combination of these factors. The MAPK inhibitor PD98059 blocked the ability of E(2) to increase intracellular prolactin concentrations. Similarly, the IGF-I receptor antagonist, JB1, blocked the effect of E(2) on prolactin synthesis and MAPK activation, as did the oestrogen receptor antagonist ICI182 780. These results suggest that, to stimulate prolactin synthesis, E(2) activates the MAPK cascade and that this requires the presence of both oestrogen and IGF-I receptors. PMID- 15796761 TI - Seasonal differences in the secretion of luteinising hormone and prolactin in response to N-methyl-DL-aspartate in starlings (Sturnus vulgaris). AB - In birds, unlike mammals, seasonal changes in reproductive function are associated with marked changes in the amount of gonadotrophin-releasing hormone (GnRH) stored in the hypothalamus. Prolonged exposure to long photoperiods leads to photorefractoriness after the breeding season. Photorefractory birds have low hypothalamic concentrations of chicken GnRH-I (cGnRH-I). Exposure to short photoperiods results in renewed cGnRH-I synthesis and increased hypothalamic stores. Birds are then photosensitive and subsequent exposure to an increase in photoperiod results in increased cGnRH-I secretion and gonadal maturation. However, it is unclear whether the reverse is true at the time of gonadal regression during long photoperiods (i.e. that a decrease in GnRH-I synthesis precedes regression). Hypothalamic stores of cGnRH-I, and possibly therefore of releasable GnRH-I, decrease after regression. Single injections of the glutamate agonist N-methyl-DL-aspartate (NMA) were used as a probe to assess releasable stores of cGnRH-I in male starlings at four physiologically different reproductive stages. Treatment induced the greatest increase in luteinising hormone (LH) in photosensitive birds in January, and a slight increase in sexually mature birds in April. There was a slight but significant increase in June, immediately after testicular regression, but no increase in fully photorefractory birds in September. These data confirm that photorefractoriness is associated with a lack of releasable cGnRH-I, but that decreased synthesis of cGnRH-I is not the proximate cause of regression. There was an increase in prolactin in response to NMA at all times. The magnitude of the response was proportional to pre-treatment concentrations, with the greatest response in June. It is suggested that high circulating prolactin may fine-tune the timing of gonadal regression in advance of the inhibition of cGnRH-I synthesis. PMID- 15796762 TI - Provocation of kainic acid receptor mRNA changes in the rat paraventricular nucleus by insulin-induced hypoglycaemia. AB - Hypoglycaemia induced by insulin injection is a powerful stimulus to the hypothalamic-pituitary-adrenal (HPA) axis and drives the secretion of corticotropin-releasing hormone and vasopressin from the neurones in the paraventricular nucleus (PVN), as well as the downstream hormones, adrenocorticotropic hormone and corticosterone. In some brain regions, hypoglycaemia also provokes increases in extracellular fluid concentrations of glutamate. Regulation of glutamatergic mechanisms could be involved in the control of the HPA axis during hypoglycaemic stress and one potential site of regulation might be at the receptors for glutamate, which are expressed in the PVN. Insulin (2.0 IU/kg, i.p.) or saline was administered to adult male Sprague Dawley rats and the animals were sacrificed 30 min, 180 min and 24 h after injection. The amount of several kainic acid-preferring glutamate receptor mRNAs (i.e. KA2, GluR5 and GluR6) were assessed in the PVN by in situ hybridisation histochemistry. Injection of insulin induced a rapid fall in plasma glucose concentrations, which was mirrored by an increase in plasma corticosterone concentrations. KA2 and GluR5 mRNAs are highly expressed within the rat PVN, and responded to hypoglycaemia with robust increases in expression that endured beyond the period of hypoglycaemia itself. However, GluR6 mRNA is expressed in the areas adjacent to the PVN and hypoglycaemic stress failed to alter expression of this mRNA. These experiments suggest that kainic acid-preferring glutamate receptors are responsive to changes in plasma glucose concentrations and may participate in the activation of the PVN neurones during hypoglycaemic stress. PMID- 15796763 TI - Clocks and the black box: circadian influences on gonadotropin-releasing hormone secretion. AB - Although the mechanisms underlying hypothalamic surge secretion of gonadotropin releasing hormone (GnRH) in rodent models have remained enduring mysteries in the field of neuroendocrinology, the identities of two fundamental constituents are clear. Elevated ovarian oestrogen, in conjunction with circadian signals, combine to elicit GnRH surges that are confined to the afternoon of the proestrus phase. The phenomenon of oestrogen positive feedback, although extensively investigated, is not completely understood, and may involve the actions of this steroid directly on GnRH perikarya, as well as on the activity of neuronal afferents. Additionally, whereas many studies have focused upon regulation of GnRH surge secretion by the neuroanatomical biological clock, the suprachiasmatic nucleus, it remains unclear why this daily signal is capable of stimulating surges only in the presence of oestrogen. This review re-examines multiple models of circadian control of reproductive neurosecretion, armed with the recent characterisation of the intracellular transcriptional feedback loops that comprise the circadian clock, and attempts to evaluate previous studies on this topic within the context of these new discoveries. Recent advances reveal the presence of oscillating circadian clocks throughout the central nervous system and periphery, including the anterior pituitary and hypothalamus, raising the possibility that synchrony between multiple cellular clocks may be involved in GnRH surge generation. Current studies are reviewed that demonstrate the necessity of functional clock oscillations in generating GnRH pulsatile secretion in vitro, suggesting that a GnRH-specific intracellular circadian clock may underlie GnRH surges as well. Multiple possible steroidal and neuronal contributions to GnRH surge generation are discussed, in addition to how these signals of disparate origin may be integrated at the cellular level to initiate this crucial reproductive event. PMID- 15796765 TI - Distribution of corticotropin-releasing factor binding protein-immunoreactivity in the rat hypothalamus: association with corticotropin-releasing factor-, urocortin 1- and vimentin-immunoreactive fibres. AB - Corticotropin-releasing factor binding protein (CRF-BP) is a 37-kDa protein with high affinity binding sites for both corticotropin-releasing factor (CRF) and urocortin 1. Previous studies have examined the distribution of CRF-BP mRNA and peptide within the central nervous system. Due to the predominant cortical localisation, very little is known about CRF-BP in subcortical structures including the hypothalamus. The present study employed immunohistochemistry to characterise the distribution of CRF-BP-like-immunoreactive (-ir) cells and fibres in the rat hypothalamus. Bipolar and multipolar CRF-BP-ir neurones were scattered throughout the rostro-caudal extent of the hypothalamus. Distinct clusters of CRF-BP-ir neurones were identified in the anterior and posterior parvocellular and dorsal cap subdivisions of the paraventricular nucleus (PVN), as well as in the dorsal hypothalamic area, dorsomedial hypothalamic nucleus (DMN), ventral premammillary nucleus and zona incerta. CRF-BP-ir fibres extending from the third ventricle were found in the mediobasal hypothalamus and within the arcuate nucleus-median eminence region. Double immunostaining together with confocal microscopy demonstrated that the CRF-BP-immunostained fibres within the mediobasal hypothalamus coincided with vimentin immunostaining indicating that CRF-BP-ir is present within tanycytes. To define the relationship between CRF-BP ir cells and endogenous ligands for CRF-BP, double immunohistochemistry was performed to examine possible sites within the hypothalamus where CRF- or urocortin 1-ir fibres innervate regions that contain CRF-BP-ir cell bodies. CRF BP-ir cell bodies typically coincided with dense CRF-ir, but not urocortin 1-ir fibre innervation. CRF-ir fibre innervation was moderate to high within the anterior and posterior parvocellular subdivisions of the PVN, the dorsal cap of the PVN, DMN and the zona incerta; all regions that contained CRF-BP-ir cell populations. These studies demonstrate that, within the hypothalamus, CRF-BP-ir cells and fibres are concentrated within a circuitry known to be involved in mediating neuroendocrine and autonomic responses to stress. PMID- 15796766 TI - Oestradiol effect on galanin-immunoreactive neurones in the diencephalon of the ewe. AB - Galanin is a neuropeptide involved in the regulation of numerous functions such as reproduction. In female rats, this peptide stimulates gonadotropin-releasing hormone (GnRH)/luteinizing hormone release and its synthesis is stimulated by oestradiol. It could therefore be an intermediary between the oestrogenic signal from the ovaries and the GnRH neurones (e.g. during the time course leading to the preovulatory GnRH surge). However, although the involvement of galanin is well-known in rodents, it is poorly understood in ewes. Using immunohistochemistry with a specific antigalanin antiserum, we detected the peptide in neurones of two groups of ovariectomized ewes treated for 6 h with subcutaneous implants, either with oestradiol (experimental group) or empty (control group). The galanin-immunoreactive neurones were counted in three areas, the preoptic area, the bed nucleus of the stria terminalis and the infundibular nucleus, using a computerized image analysis system. There was no change in the mean number of galanin-immunoreactive (GAL-ir) neurones in the infundibular nucleus (37 +/- 12 neurones/section in treated animals and 31 +/- 11 in controls) or in the bed nucleus of the stria terminalis (22 +/- 5 neurones/section in treated animals and 16 +/- 4 in controls), but the number of GAL-ir neurones was higher in the preoptic area in treated than in control ewes (35 +/- 4 versus 14 +/- 10, P < 0.001). To determine whether the neurones of the preoptic area were directly sensitive to oestradiol, we performed double immunohistochemical labelling for oestradiol receptor alpha and galanin. More than 50% of the GAL-ir neurones contained the oestradiol receptor alpha and therefore could be directly regulated by oestradiol. These results indicate that oestradiol might act directly on a GAL-ir neuronal population situated in the preoptic area, without any effect on the GAL-ir neurones of the infundibular nucleus or the bed nucleus of the stria terminalis. Because a 6-h oestradiol treatment can induce a preovulatory GnRH surge in ewes, the GAL-ir neuronal population of the preoptic area might be one of the neuronal systems by which oestradiol activates the GnRH neurones. However, although the morphological relationships between galanin and GnRH neurones have been described in rodents, they remain to be demonstrated in the ewe. PMID- 15796767 TI - Bacterial endotoxin (lipopolysaccharide) stimulates interleukin-6 production and inhibits growth of pituitary tumour cells expressing the toll-like receptor 4. AB - Members of the Toll receptor (Tlr) family have a crucial role in the innate immune response following bacterial infection. The effects of Gram-negative bacteria-derived endotoxins (lipopolysaccharide, LPS) are predominantly mediated by Tlr4, and we have recently shown that pituitary folliculostellate cells express functional Tlr4. In the present study, we investigated whether Tlr4 is also present in normal and transformed endocrine epithelial pituitary cell types. By reverse transcriptase-polymerase chain reaction, Tlr4 mRNA expression was found in some pituitary epithelial tumour cell lines (AtT20, HP75), whereas others were negative (GH3, alphaT3-1). Tlr4 protein was detected by immunohistochemistry in a few epithelial cells in normal human anterior pituitaries and in 26 out of 67 human pituitary tumours analysed. LPS had no effect on adrenocorticotropic hormone secretion in Tlr4-positive AtT20 cells, but it suppressed the growth of these cells in a dose-dependent manner. As expected, neither hormone secretion, nor growth of Tlr4-negative GH3 cells was affected by LPS. In cell cultures of Tlr4-positive pituitary adenomas, LPS dose-dependently stimulated the production of interleukin (IL)-6, which is known to induce growth and hormone production in pituitary tumours. The LPS-induced IL-6 production was blocked by the specific p38alphaMAP kinase inhibitor, SB203580, and by the synthetic glucocorticoid, dexamethasone. The data suggest that, during Gram negative bacteria-induced infections or inflammatory processes, LPS could affect pituitary tumour pathophysiology and progression in the subset of Tlr4-expressing pituitary adenomas. PMID- 15796768 TI - Progesterone can block the preovulatory gonadotropin-releasing hormone/luteinising hormone surge in the ewe by a direct inhibitory action on oestradiol-responsive cells within the hypothalamus. AB - Elevated oestradiol concentrations during the follicular phase stimulate a surge in gonadotropin-releasing hormone (GnRH) and luteinising hormone (LH) concentrations, which leads to ovulation. Progesterone can block the oestradiol induced GnRH/LH surge, but the mechanism that is involved is unclear. We examined the effect of progesterone on oestradiol-induced activation of cells within the ovine hypothalamus/preoptic area (POA) to determine: (i) in which regions progesterone acts to block the GnRH/LH surge and (ii) whether progesterone directly or indirectly prevents activation of oestradiol-responsive cells. Cellular activation was assessed by measuring the number of cells that expressed Fos (an immediate early gene). Exposure to increased oestradiol concentrations in the absence of progesterone (which normally stimulates a LH surge) did not cause any region-specific changes in hypothalamic Fos expression during the activation stage of the LH surge-induction process (Experiment 1). The same treatment significantly increased cellular activation within the POA, lateral septum (LS), and arcuate nucleus at the time of surge onset (Experiment 2). Concurrent exposure to increased oestradiol and progesterone concentrations during the activation stage of the surge-induction process (which normally blocks the LH surge) was associated with significantly reduced cellular activation within the ventromedial hypothalamus and anterior hypothalamic area, relative to the positive controls (oestradiol increment alone) and arcuate nucleus relative to the negative controls (no increment in oestradiol) during the activation stage (Experiment 1). At the time of surge onset (Experiment 2), exposure to progesterone during the activation period prevented the oestradiol-induced increase in cellular activation that occurred in the POA, LS and arcuate nucleus of the positive controls. These results demonstrated that oestradiol and progesterone induced differential region- and time-specific effects on cellular activation within the regions of the ovine brain that generate the preovulatory GnRH/LH surge. Moreover, the lack of cellular activation within the POA, LS and arcuate nucleus at the time of surge onset in animals exposed to progesterone during the activation stage is consistent with the hypothesis that progesterone can block the preovulatory surge by direct inhibition of oestradiol-induced cellular activation in these areas. PMID- 15796769 TI - The human MT1 melatonin receptor stimulates cAMP production in the human neuroblastoma cell line SH-SY5Y cells via a calcium-calmodulin signal transduction pathway. AB - Melatonin regulates circadian and seasonal physiology via melatonin receptors expressed in the brain. However, little is known about the signal transduction mechanisms that mediate the action of melatonin in neuronal cells. To begin to address this issue, we expressed the human MT(1) receptor in the human neuroblastoma SH-SY5Y cell line. In this cell line, melatonin acutely stimulated cAMP synthesis through a calcium-calmodulin dependent pathway. This stimulatory effect was independent of an interaction with G(i) or G(s) G proteins and dependent upon internal calcium stores. Melatonin also potentiated forskolin activated cAMP synthesis. Differentiation of the neuroblastoma cells with retinoic acid to the neuronal phenotype did not alter the ability of melatonin to acutely stimulate cAMP. These data may be relevant to the neuronal action of melatonin and highlight the importance of the cellular context of expression of melatonin and other G protein-coupled receptors. PMID- 15796770 TI - Oestrogen and testosterone modulate the firing activity of dorsal raphe nucleus serotonergic neurones in both male and female rats. AB - Women are twice as likely to suffer from mood disorders than men. Moreover, a growing body of evidence suggests a reciprocal modulation between sex steroids and the serotonin (5-HT) system. A previous study from our laboratory has shown that the progesterone metabolites 5beta-pregnane-3,20-dione (5beta-DHP) and 5alpha-pregnan-3alpha-ol,20-one (3alpha,5alpha-THP), as well as dehydroepiandrosterone (DHEA), increase the firing activity of dorsal raphe nucleus (DRN) 5-HT neurones in female rats. The present study was undertaken to assess the effects of these steroids in male rats, as well as the effects of testosterone and 17beta-oestradiol (17beta-E) in both sexes, and finally to evaluate gender differences in the modulation of the 5-HT neuronal firing activity by these different neuroactive steroids. Male rats were treated i.c.v., for 7 days, with a dose of 50 microg/kg/day of one of the following steroids: progesterone, 5beta-DHP, 3alpha,5alpha-THP, DHEA, testosterone, 17beta-hydroxy 5alpha-androstan-3-one (5alpha-DHT) and 17beta-E. Some rats also received a 3-day administration of testosterone (50 microg/kg/day, i.c.v). Females were treated in the same fashion with testosterone and 17beta-E. Extracellular unitary recordings of 5-HT neurones, obtained in vivo in the DRN of these rats, revealed that testosterone and 17beta-E increased the firing activity of 5-HT neurones in both males and females. In males, the effect of testosterone could already be seen after 3 days of treatment. Neither castration nor any treatment with other steroids significantly modified the firing rate of male 5-HT neurones. Taken together with previous findings, the results of the present study indicate both similarities and differences between sexes in the modulation of 5-HT neurones by some steroids. This could prove important in understanding gender differences in mood disorders. PMID- 15796771 TI - Hypothalamic STAT proteins: regulation of somatostatin neurones by growth hormone via STAT5b. AB - Signal transducers and activators of transcription (STATs) are a family of transcription factors linked to class I cytokine receptors. In the present study, we investigated whether their distribution in the hypothalamus reflects the feedback regulation by growth hormone and what role they might play in the functioning of target neurones. We demonstrate that each of the seven known STATs has a distinct distribution in the hypothalamus. Notably, the STAT5 proteins, that are important in growth hormone (GH) and prolactin signalling in peripheral tissues, were expressed in somatostatin neurones of the periventricular nucleus and dopamine neurones of the arcuate nucleus. Because somatostatin neurones are regulated by feedback from circulating GH, we investigated the importance of STAT5 in these neurones. We demonstrate that STAT5b protein expression, similar to somatostatin mRNA, is sexually dimorphic in the periventricular nucleus of rats and mice. Furthermore, chronic infusion of male dwarf rats with GH increased the expression of STAT5b, while a single injection of GH into similar rats induced the phosphorylation of STAT5 proteins. The cellular abundance of somatostatin mRNA in STAT5b-deficient mice was significantly reduced in the periventricular nucleus, effectively reducing the sexually dimorphic expression. These results are consistent with the hypothesis that STAT5 proteins are involved in the feedback regulation of somatostatin neurones by GH, and that these neurones may respond to patterned GH secretion to reinforce sexual dimorphism in the GH axis. PMID- 15796772 TI - EMBASE search strategies for identifying methodologically sound diagnostic studies for use by clinicians and researchers. AB - BACKGROUND: Accurate diagnosis by clinicians is the cornerstone of decision making for recommending clinical interventions. The current best evidence from research concerning diagnostic tests changes unpredictably as science advances. Both clinicians and researchers need dependable access to published evidence concerning diagnostic accuracy. Bibliographic databases such as EMBASE provide the most widely available entree to this literature. The objective of this study was to develop search strategies that optimize the retrieval of methodologically sound diagnostic studies from EMBASE for use by clinicians. METHODS: An analytic survey was conducted, comparing hand searches of 55 journals with retrievals from EMBASE for 4,843 candidate search terms and 6,574 combinations. All articles were rated using purpose and quality indicators, and clinically relevant diagnostic accuracy articles were categorized as 'pass' or 'fail' according to explicit criteria for scientific merit. Candidate search strategies were run in EMBASE, the retrievals being compared with the hand search data. The proposed search strategies were treated as "diagnostic tests" for sound studies and the manual review of the literature was treated as the "gold standard." The sensitivity, specificity, precision and accuracy of the search strategies were calculated. RESULTS: Of the 433 articles about diagnostic tests, 97 (22.4%) met basic criteria for scientific merit. Combinations of search terms reached peak sensitivities of 100% with specificity at 70.4%. Compared with best single terms, best multiple terms increased sensitivity for sound studies by 8.2% (absolute increase), but decreased specificity (absolute decrease 6%) when sensitivity was maximized. When terms were combined to maximize specificity, the single term "specificity.tw." (specificity of 98.2%) outperformed combinations of terms. CONCLUSION: Empirically derived search strategies combining indexing terms and textwords can achieve high sensitivity and specificity for retrieving sound diagnostic studies from EMBASE. These search filters will enhance the searching efforts of clinicians. PMID- 15796774 TI - International Retrovirology Association brings together scientists and clinicians to bridge discoveries about human T-lymphotropic viruses from the laboratory to clinical trials. AB - Human T-lymphotropic virus type 1 (HTLV-1) and HTLV-2 were among the first human retroviruses discovered in the early 1980's. The International Retrovirology Association is an organized effort that fostered the efforts of scientists and clinicians to form interdisciplinary groups to study this group of retroviruses and their related diseases. The Association promotes excellent science, patient education, and fosters the training of young scientists to promote "bench-to bedside" research. The International Conference on Human Retrovirology: HTLV and Related Viruses sponsored by the Association supports clinicians and researchers in the exchange of research findings and stimulation of new research directions. This years conference will be held from June 22 to 25, in Montego Bay, Jamaica http://www.htlvconference.org.jm/. Since its inception in 1988, these conferences have provided a highly interactive forum for the global community of HTLV scientists. This is of particular importance as HTLV research enters its third decade and a new generation of scientists takes over this important work. Many of the scientists attending the meeting will be from developing countries where HTLV is endemic, consistent with the history of international collaborations that have characterized HTLV research. The International Conference on Human Retrovirology provides a unique opportunity for researchers of all disciplines interested in HTLV infections to meet their peers and to address the questions facing clinicians and scientists who study retroviruses, like HTLV. PMID- 15796773 TI - A hypothesis-generating study to evaluate platelet activity in diabetics with chronic kidney disease. AB - BACKGROUND: It is well described that diabetes mellitus is a hypercoagulable state. It is also known that patients with renal dysfunction have impaired platelet aggregation and function. It is not well described how renal dysfunction affects the hypercoagulability associated with diabetes. This post-hoc sub-group analysis compares platelet function, clot structure and thrombin generation time at baseline, and following enoxaparin exposure in three groups of subjects. METHODS: 30 total subjects were evaluated in the three groups: Group I: normal controls (n = 10), Group II: subjects with renal dysfunction but without diabetes (n = 13), and Group III: subjects with concomitant diabetes and renal dysfunction (n = 7). For each subject, platelet contractile force (PCF), clot elastic modulus (CEM) and thrombin generation time (TGT) were simultaneously measured in whole blood at baseline, and following increasing enoxaparin antifactor Xa activity exposure. The group means for each parameter were determined and compared using one-way analysis of variance, with post-hoc Tukey-Kramer test. RESULTS: At baseline, subjects in Group III (diabetics with concomitant renal dysfunction) display significantly enhanced platelet activity, as measured by PCF (p = 0.003) and CEM (p = 0.03), relative to the non-diabetic Groups I and II. Subjects in Group II (renal dysfunction without diabetes) had significantly prolonged TGT values relative to controls when the antifactor Xa activity concentration reached 0.5 (p = 0.007), 1.0 (p = 0.005) and 3.0 IU/mL (p < 0.0001), respectively. There were no differences between Group II and Group III with respect to TGT at these antifactor Xa activity concentrations. When the antifactor Xa activity concentration reached 3.0 IU/mL, Groups II and III formed significantly less rigid blood clots (CEM p = 0.003) and also trended toward reduced PCF (p = 0.06) relative to Group I. CONCLUSION: This hypothesis-generating sub-group analysis suggests that at baseline, patients with concomitant diabetes and renal dysfunction have significantly enhanced platelet activity (PCF), and form more rigid blood clots (CEM) compared to controls and subjects with renal dysfunction but no diabetes. This may suggest that the presence of renal dysfunction does not ameliorate the hypercoagulable state associated with diabetes. Secondly, it appears that subjects with renal dysfunction but without diabetes have an enhanced response to enoxaparin relative to controls. PMID- 15796775 TI - Internal fixation of femoral shaft fractures in children by intramedullary Kirschner wires (a prospective study): its significance for developing countries. AB - BACKGROUND: To evaluate internal fixation by intramedullary Kirschner wires as a surgical technique in the treatment of femoral shaft fractures in children by a prospective study. METHODS: 17 femoral shaft fractures at various levels in 16 children aged 2-15 years were treated by closed intramedullary Kirschner wiring under image intensifier control between May 2000 and October 2003. No external splint was used. RESULTS: Fracture union was achieved in 6-14 weeks. Non-weight bearing crutch walking was started 2-3 days after surgery. Full weight bearing started 6-14 weeks. Average operative time was 40 min (range 20-72 min). Wires were removed after 8-22 weeks. There were no infections, no limb length disparity. One child had pin track ulceration. A big child of 14 years had angulation of the fracture. CONCLUSION: Intramedullary nailing of femoral shaft fractures in children by stainless steel Kirschner wires is an effective method, which compares well with other studies. It is a simple procedure, which can be easily reproduced. Blood loss is minimal, and the operative time short. There is no need pre-bend the wires in a C or S curve. Stainless steel Kirschner wires are cheap, universally available, and can be manufactured locally. The cost of Image intensifiers is affordable in most of the cities of the developing countries. The hospital does not have to maintain a costly inventory. Provides early mobility, return to home and, school. Gives a predictable clinical pathway and reduces occupancy of hospital beds. The technique was successfully applied for internal fixation of other diaphyseal fractures in children and some selected diaphyseal fractures in adults. Based on my experience and a review of the literature, I recommend this technique as a modality for treatment of femoral shaft fractures in children aged 2 to 14 years. PMID- 15796776 TI - Amplifier spurious input current components in electrode-electrolyte interface impedance measurements. AB - BACKGROUND: In Impedance Microbiology, the time during which the measuring equipment is connected to the bipolar cells is rather long, usually between 6 to 24 hrs for microorganisms with duplication times in the order of less than one hour and concentrations ranging from 10(1) to 10(7) [CFU/ml]. Under these conditions, the electrode-electrolyte interface impedance may show a slow drift of about 2%/hr. By and large, growth curves superimposed on such drift do not stabilize, are less reproducible, and keep on distorting all over the measurement of the temporal reactive or resistive records due to interface changes, in turn originated in bacterial activity. This problem has been found when growth curves were obtained by means of impedance analyzers or with impedance bridges using different types of operational amplifiers. METHODS: Suspecting that the input circuitry was the culprit of the deleterious effect, we used for that matter (a) ultra-low bias current amplifiers, (b) isolating relays for the selection of cells, and (c) a shorter connection time, so that the relays were maintained opened after the readings, to bring down such spurious drift to a negligible value. Bacterial growth curves were obtained in order to test their quality. RESULTS: It was demonstrated that the drift decreases ten fold when the circuit remained connected to the cell for a short time between measurements, so that the distortion became truly negligible. Improvement due to better-input amplifiers was not as good as by reducing the connection time. Moreover, temperature effects were insignificant with a regulation of +/- 0.2 [ degrees C]. Frequency did not influence either. CONCLUSION: The drift originated either at the dc input bias offset current (Ios) of the integrated circuits, or in discrete transistors connected directly to the electrodes immersed in the cells, depending on the particular circuit arrangement. Reduction of the connection time was the best countermeasure. PMID- 15796777 TI - CGMIM: automated text-mining of Online Mendelian Inheritance in Man (OMIM) to identify genetically-associated cancers and candidate genes. AB - BACKGROUND: Online Mendelian Inheritance in Man (OMIM) is a computerized database of information about genes and heritable traits in human populations, based on information reported in the scientific literature. Our objective was to establish an automated text-mining system for OMIM that will identify genetically-related cancers and cancer-related genes. We developed the computer program CGMIM to search for entries in OMIM that are related to one or more cancer types. We performed manual searches of OMIM to verify the program results. RESULTS: In the OMIM database on September 30, 2004, CGMIM identified 1943 genes related to cancer. BRCA2 (OMIM *164757), BRAF (OMIM *164757) and CDKN2A (OMIM *600160) were each related to 14 types of cancer. There were 45 genes related to cancer of the esophagus, 121 genes related to cancer of the stomach, and 21 genes related to both. Analysis of CGMIM results indicate that fewer than three gene entries in OMIM should mention both, and the more than seven-fold discrepancy suggests cancers of the esophagus and stomach are more genetically related than current literature suggests. CONCLUSION: CGMIM identifies genetically-related cancers and cancer-related genes. In several ways, cancers with shared genetic etiology are anticipated to lead to further etiologic hypotheses and advances regarding environmental agents. CGMIM results are posted monthly and the source code can be obtained free of charge from the BC Cancer Research Centre website http://www.bccrc.ca/ccr/CGMIM PMID- 15796779 TI - BCG skin reaction in Mantoux-negative healthy children. AB - BACKGROUND: Tuberculosis poses a great challenge, especially in children. The response of BCG Test may be different in previously vaccinated children and needs to be considered before interpreting positivity for TB. This study has been carried out to determine the pattern of BCG reaction comparing previously vaccinated with non-vaccinated children. METHODS: The study was conducted in the healthy school children aged 4-6 years. The BCG skin reaction in Mantoux-negative children was compared between children with and without previous BCG scar. After the Mantoux and BCG Test, the analysis of variance was done as per protocol. RESULTS: Out of 50 children previously BCG vaccinated, 39(78%) showed exaggerated BCG test responses while out of another 50 children who were not vaccinated for TB, only 9(18%) showed exaggerated BCG Test response (p-value < 0.00001). Average induration obtained in children who were immunized with BCG at birth was much greater than those who were not immunized. 80% and 76% males and females respectively in Group I showed exaggerated BCG response while 16% and 20% males and females respectively of Group II showed exaggerated BCG response. CONCLUSION: The present study indicates that normal healthy children may have a mild exaggerated BCG Test response i.e. induration up to 8 mm because of prior BCG vaccination. Therefore, BCG Test, though important should not be the only criteria for start of chemotherapy for TB in children as the side effects of drugs may cause much morbidity. An induration up to 8 mm after the BCG Test can be normal in Indian settings due to exposure to Mycobacterium in environment and/or BCG vaccine. PMID- 15796778 TI - Imaging plant cell death: GFP-Nit1 aggregation marks an early step of wound and herbicide induced cell death. AB - BACKGROUND: A great deal is known about the morphological endpoints of plant cell death, but relatively little is known about its sequence of events and/or its execution at the biochemical level. Live cell imaging using GFP-tagged markers is a powerful way to provide dynamic portraits of a cellular process that can in turn provide a descriptive foundation valuable for future biochemical and genetic investigations. RESULTS: While characterizing a collection of random GFP-protein fusion markers we discovered that mechanical wounding induces rapid aggregation of a GFP-Nitrilase 1 fusion protein in Arabidopsis cells directly abutting wound sites. Time-lapse imaging of this response shows that the aggregation occurs in cells that subsequently die 30-60 minutes post-wounding, indicating that GFP-Nit1 aggregation is an early marker of cell death at wound sites. Time-lapse confocal imaging was used to characterize wound-induced cell death using GFP-Nit1 and markers of the nucleus and endoplasmic reticulum. These analyses provide dynamic portraits of well-known death-associated responses such as nuclear contraction and cellular collapse and reveal novel features such as nuclear envelope separation, ER vesiculation and loss of nuclear-lumen contents. As a parallel system for imaging cell death, we developed a chemical method for rapidly triggering cell death using the herbicides bromoxynil or chloroxynil which cause rapid GFP-Nit1 aggregation, loss of nuclear contents and cellular collapse, but not nuclear contraction, separating this response from others during plant cell death. CONCLUSION: Our observations place aggregation of Nitrilase 1 as one of the earliest events associated with wound and herbicide-induced cell death and highlight several novel cellular events that occur as plant cells die. Our data create a detailed descriptive framework for future investigations of plant cell death and provide new tools for both its cellular and biochemical analysis. PMID- 15796782 TI - Practical considerations for estimating clinical trial accrual periods: application to a multi-center effectiveness study. AB - BACKGROUND: Adequate participant recruitment is vital to the conduct of a clinical trial. Projected recruitment rates are often over-estimated, and the time to recruit the target population (accrual period) is often under-estimated. METHODS: This report illustrates three approaches to estimating the accrual period and applies the methods to a multi-center, randomized, placebo controlled trial undergoing development. RESULTS: Incorporating known sources of accrual variation can yield a more justified estimate of the accrual period. Simulation studies can be incorporated into a clinical trial's planning phase to provide estimates for key accrual summaries including the mean and standard deviation of the accrual period. CONCLUSION: The accrual period of a clinical trial should be carefully considered, and the allocation of sufficient time for participant recruitment is a fundamental aspect of planning a clinical trial. PMID- 15796781 TI - Identification and characterization of Iporin as a novel interaction partner for rab1. AB - BACKGROUND: The small GTPase rab1a and its isoform rab1b are essential regulating components in the vesicle transport between the ER and the Golgi apparatus. Rab1 is thought to act as a molecular switch and can change between an active GTP bound and an inactive GDP-bound conformation. To elucidate the function of rab1, several approaches have been established to isolate effector proteins, which interact with the activated conformation of rab1. To date p115, GM130, golgin-84 and MICAL have been identified as direct interacting partners. Together with rab1, these molecules are components of a protein complex, which mediates and regulates intracellular vesicle transport. RESULTS: Here, we report the characterization of Iporin, which is similar to KIAA0375 as a novel rab1 interacting protein. It was initially identified by yeast two-hybrid screening experiments with the active mutant of rab1b (rab1b Q67R) as bait. Iporin contains a SH3 domain and two polyproline stretches, which are known to play a role in protein/protein interactions. In addition, Iporin encloses a RUN domain, which seems to be a major part of the rab1binding domain (R1BD). Iporin is ubiquitously expressed and immunofluorescence staining displays a cytosolic punctual distribution. Interestingly, we also show that Iporin interacts with another rab1 interacting partner, the GM130 protein. CONCLUSION: Our results demonstrate that Iporin is a potential new interacting partner of rab1. Iporin is different from already identified rab1 interacting proteins concerning protein structure and cellular localization. We conclude that Iporin might function as a link between the targeting of ER derived vesicles, triggered by the rab1 GTPase and a signaling pathway regulated by molecules containing SH3 and/or poly-proline regions. The characterization of this novel intermolecular relation could help to elucidate how vesicles find their way from ER to the Golgi apparatus. PMID- 15796783 TI - Chronic tendon pathology: molecular basis and therapeutic implications. AB - Tendons are frequently affected by chronic pain or rupture. Many causative factors have been implicated in the pathology, which until relatively recently was under-researched and poorly understood. There is now a greater knowledge of the molecular basis of tendon disease. Most tendon pathology (tendinopathy) is associated with degeneration, which is thought to be an active, cell-mediated process involving increased turnover and remodelling of the tendon extracellular matrix. Degradation of the tendon matrix is mediated by a variety of metalloproteinase enzymes, including matrix metalloproteinases and 'aggrecanases'. Neuropeptides and other factors released by stimulated cells or nerve endings in or around the tendon might influence matrix turnover, and could provide novel targets for therapeutic intervention. PMID- 15796784 TI - Readiness potential and movement initiation in the rat. AB - Cortical field potentials were recorded by electrodes implanted chronically on the surface and at a 2.0 mm depth in various cortical areas in the left hemisphere in the rat during self-paced movements of the right forelimb. A surface-negative (s-N), depth-positive (d-P) cortical field potential appeared about 1.0 s (range: 0.5-1.5 s) before movement onset in the rostral (RFA) and caudal (CFA) forelimb areas of the motor cortex, and the somatosensory cortex, but not in the occipital cortex. Bipolar recording of electromyographic activities induced by the electrical stimulation of various cortical loci was also performed by pairs of steel electrodes inserted in the face, trunk, forelimb and hindlimb muscles on both sides. The stimulation of the forelimb motor cortex activated the face and/or forelimb muscles, while that of the somatosensory cortex generally activated several body part muscles including the forelimb muscle. Stronger stimulus intensity was requested to elicit the activities of most of the ipsilateral muscles to the cortex stimulated than the contralateral ones. The minimum intensity for inducing the forelimb muscle activity was lowest in the CFA among cortical areas producing the activity. The stimulation of cortical loci in which the s-N, d-P potential was recorded could induce muscle activities in the forelimb contralateral to the stimulation. It is suggested that the s-N, d-P potential is the readiness potential for activating muscles to initiate movement in the rat forelimb. PMID- 15796785 TI - Vestibulo-cardiorespiratory responses at the onset of chair rotation in endurance runners. AB - Stimulation of the vestibular system has been reported to elicit ventilatory and circulatory changes in humans. The purpose of this study was to clarify the characteristics of vestibular-mediated ventilatory and circulatory responses in male endurance runners at the onset of passive chair rotation, which selectively stimulates the semicircular canals. Fourteen runners and 14 male untrained subjects participated. The vestibular stimulus test, which consists of 180 degrees chair rotations (left or right half-turns on an earth-vertical axis) for a duration of 2 s, was carried out on each subject. Inspiratory minute ventilation, tidal volume, respiratory frequency, heart rate, and blood pressure were measured by breath-by-breath and beat-to-beat techniques before, during, and after the chair rotation for a total of 60 s. It was found in this study that (i) the relative change of minute ventilation response in the endurance runners was significantly (P < 0.05) greater than in the untrained subjects during and after the rotation, and that (ii) no significant group differences were observed in heart rate and mean blood pressure responses during and after the rotation. In conclusion, vestibular-mediated ventilatory response, but not circulatory response, at the onset of the chair rotation in the endurance runners was significantly greater than that in the untrained subjects. The results from the present study suggest that an increase in vestibulo-ventilatory response would be attributed to an adaptation to long-term endurance training. PMID- 15796786 TI - Alterations in the rheological flow profile in conduit femoral artery during rhythmic thigh muscle contractions in humans. AB - The present study examined the rheological blood velocity profile in the conduit femoral artery during rhythmic muscle contractions at different muscle forces. Eight healthy volunteers performed one-legged, dynamic knee-extensor exercise at work rates of 5, 10, 20, 30, and 40 W at 60 contractions per minute. The time and space-averaged, amplitude-weighted mean (V(mean)) and maximum (V(max)) blood flow velocities in the common femoral artery were measured during the cardiosystolic phase (CSP) and cardiodiastolic phase (CDP) by the Doppler ultrasound technique. The V(max)/V(mean) ratio was used as a flow profile index, in which a ratio of approximately 1 indicates a "flat velocity flow profile" and a ratio significantly >1 indicates a "parabolic velocity flow profile." At rest, the V(max)/V(mean) ratio was approximately 1.3 and approximately 1.8 during the CSP and CDP, respectively. The V(max)/V(mean) ratio was higher (p < 0.01) during the CDP than during the CSP, both at rest and at all work rates. The V(max)/V(mean) ratio during the CSP was higher (p < 0.01) at 30 and 40 W compared to at rest. The V(max)/V(mean) ratio during the CDP was lower (p < 0.05) at 5 and 10 W compared to at rest. There was a positive linear correlation between blood flow and incremental work rates during both the CSP and CDP, respectively. Thus under resting conditions, the findings indicate a "steeper" parabolic velocity profile during the CDP than during the CSP. The velocity profile during the CDP furthermore shifts to being less "steep" during rhythmic muscle contractions at lower intensities, but to being reelevated and normalized as at rest during higher intensities. The "steepness" of the parabolic velocity profile observed during the CSP at rest increased during muscle contraction at higher intensities. In conclusion, the blood velocity in the common femoral artery is parabolic both at rest and during exercise for both the CSP and CDP, indicating the persistence of laminar flow. The occurrence of any temporary slight disturbance or turbulence in the flow at the sight of measurement in the common femoral artery does consequently not induce a persisting "disturbed" and fully flat "plug-like" velocity profile. Instead, the "steepness" of the parabolic velocity profile is only slightly modified, whereby blood flow is not impaired. Thus the blood velocity profile, besides being influenced by the muscle contraction-relaxation induced mechanical "impedance," seems also to be modulated by the cardiac- and blood pressure-phases, consequently influencing the exercise blood flow response. PMID- 15796787 TI - Effects of habitual moderate exercise on response processing and cognitive processing in older adults. AB - We examined the effects of habitual moderate exercise on central information processing in older individuals using the reaction time (RT) and P3 component of event-related brain potentials (ERP). The present study was designed to assess cognitive function by comparing groups of 20 older individuals (69.20 +/- 1.3 years active group) who regularly engage in moderate physical activity with 20 subjects (66.90 +/- 1.1 years inactive group) who do comparatively little exercise. Subjects performed a somatosensory oddball task composed of pressing a button with their right foot as fast as possible following an electrical stimulus applied to the right index finger, and not responding to an electrical stimulus applied to the left index finger. Electroencephalogram (EEG) was recorded at the frontal (Fz), central (Cz), and parietal (Pz) sites according to the International 10-20 system referenced to linked earlobes. The RT was faster for the active group than for the inactive group, and the P3 amplitude of the active group was significantly larger than that of the inactive group. Moreover, the P3 amplitude for the active group was maximum at Pz and significantly larger than at Fz and Cz, but for the inactive group it was identical between Fz and Pz. The results suggest that habitual moderate exercise exerts positive influences in older adults not only on response processing, but also on cognitive processing. PMID- 15796788 TI - Beat-to-beat modulation of atrioventricular conduction during dynamic exercise in humans. AB - A complex balance between extrinsic neural and intrinsic mechanisms is responsible for regulating atrioventricular (AV) conduction. We hypothesized that atrial excitation interval is shortened during dynamic exercise by extrinsic cardiac autonomic activity and that if AV conduction time responds inversely to fluctuation in atrial rhythm, ventricular excitation interval will be maintained at the predetermined cardiac cycle length. To examine such inverse relationship between PP interval and the subsequent change in PR interval (DeltaPR), we analyzed the beat-to-beat changes in PP, PR, and RR intervals during stair stepping exercise for 10 min in 11 sedentary and 9 trained subjects. In the sedentary group, the average PR interval significantly shortened during exercise, in parallel with the reduction in the average PP and RR intervals. The variance of PP and RR intervals was also significantly decreased during exercise. The reduction in the variance of RR interval was, however, much greater than that of PP interval, implying that AV conduction time changes inversely to fluctuation in atrial excitation rhythm. Indeed, the variance of PR interval was augmented during exercise and there was a clear inverse relationship between PP and DeltaPR intervals. Although trained subjects were characterized by their lower heart rate response during dynamic exercise, the responses in the variability of PP, PR, and RR intervals were fundamentally identical with those in sedentary subjects. We conclude that the AV nodal mechanism that operates at a higher level of heart rate during dynamic exercise may cancel fluctuation in atrial excitation interval and keep ventricular excitation rhythm at the predetermined cardiac cycle length. PMID- 15796790 TI - Location of the neuromuscular junction of the human masseter muscle estimated from the low frequency component of the surface electromyogram. AB - The position of the neuromuscular junction of the human masseter muscle has estimated from the low frequency component of the surface electromyogram. Monopolar surface electromyograms were recorded in response to clenching from eight sites with the reference electrode placed on the tip of the nose in six healthy male subjects. Component of the slow wave was separated from the raw recordings using digital filter, and the difference of the polarity and magnitude with the sites was examined. The base line of the recordings deflected either negatively or positively depending on the recording sites. These deflections coincided with bursts of muscular action potentials. Deflection of the slow wave component was largely negative over the inferior masseter close to the mandibular ramus but shifted to positive over the superior part. The greatest magnitude of the muscular action potential coincided with the most negative deflection of the slow wave. The phase of the muscular action potential burst also reversed near the site of the polarity change of the slow wave. These findings suggest that the slow wave, as detected by this method, originates mainly from the synaptic potential at the meuromuscular junction and that this is located on the inferior part of the muscle close to the mandibular ramus. PMID- 15796791 TI - Change in intrathoracic pressure in rats with spontaneous and controlled ventilation during microgravity by parabolic flight. AB - We previously reported that the intrathoracic pressure (ITP) decreases and the transmural pressure of the aortic wall (TMP) increases during 4.5 s of microgravity (muG) induced by free drop. To examine the ITP response to a longer period of muG in the absence of the respiratory rate (RR) decrease, i.e., bradypnea, which occurs at the onset of muG, we measured the aortic blood pressure at the diaphragma level (AP) and ITP. We then calculated the TMP at the aortic arch level during 20 s of muG induced by parabolic flight in anesthetized rats (n = 7) with either spontaneous ventilation (SPN-V) or controlled ventilation (CONT-V). In the SPN-V group, the bradypnea was observed in all rats after the onset of the muG (RR change -13.9 +/- 2.9/min). The ITP during muG ( 9.3 +/- 0.9 mmHg) was significantly lower than that during 1 G (-7.7 +/- 0.9 mmHg), and the TMP was significantly increased during muG (112 +/- 6 mmHg) compared to 1 G (103 +/- 5 mmHg). Similar changes in ITP and TMP were observed in the CONT-V group: During muG and 1G, respectively, the ITP was -8.4 +/- 0.6 mmHg and -5.9 +/- 0.7 mmHg, and the TMP was 112 +/- 6 mmHg and 101 +/- 6 mmHg, whereas no change in RR was observed because of the controlled ventilation. These results show that the ITP decreases and the TMP increases during muG, and they are not affected by a disturbance of respiratory rhythm. PMID- 15796794 TI - [Unite and devote to the works of pediatric clinical virology]. PMID- 15796792 TI - Vasodilatation of the hindquarters induced by antagonism of GABA(A) receptors in the freely moving rat. AB - An intracisternal injection of the GABA(A) receptor antagonist bicuculline into conscious rats increased arterial pressure and decreased hindquarter resistance. Propranolol attenuated only the resistance response, but ganglionic block abolished both responses. These findings suggest that central GABA(A) receptor blockade induces an autonomic pressor action with hindquarter vasodilatation. PMID- 15796789 TI - Endolymphatic perfusion with EGTA-acetoxymethyl ester inhibits asphyxia- and furosemide-induced decrease in endocochlear potential in guinea pigs. AB - We examined the effect of the Ca(2+) concentration in the endolymph ([Ca](e)) or in the endolymphatic surface cells ([Ca](i)) on the endocochlear potential (EP) by using an endolymphatic or perilymphatic perfusion technique, respectively. (i) A large increase in [Ca](e) up to approximately 10(-3) M with a fall in the EP was induced by transient asphyxia ( approximately 2 min) or by the intravenous administration of furosemide (60 mg/kg), and a significant correlation was obtained between the EP and p[Ca](e) (= -log [Ca](e), r = 0.998). (ii) Perfusion of the endolymph with 10 mM EGTA for 5 min neither produced any significant change in the EP nor altered the asphyxia-induced change in EP (DeltaEP(asp)), suggesting that neither [Ca](e) nor the Ca(2+) concentration gradient across the stria vascularis contributed directly to the generation of the EP in the condition of low [Ca](e). In contrast, endolymphatic perfusion with high Ca(2+) (more than 10 mM) produced a decrease in EP and a significant correlation was obtained between the EP and the Ca(2+) concentration of perfusion solution (r = 0.982), suggesting that Ca(2+) permeability may exist across the stria vascularis. (iii) The administration of a Ca(2+) chelator, EGTA-acetoxymethyl ester (AM, 0.3 mM), to the endolymph, which produced a gradual increase in EP, suppressed significantly, by 60-80%, DeltaEP(asp) or furosemide-induced changes in EP. In contrast, perilymphatic administration of 0.5 mM EGTA-AM caused no significant suppression of the DeltaEP(asp). These findings suggest that [Ca](i) plays an important role in generating/maintaining a large positive EP. PMID- 15796795 TI - [Pay special attention to research on gastrointestinal motility and functional disorders]. PMID- 15796796 TI - [Genotyping of major outer membrane protein gene of Chlamydia trachomatis by cleavase fragment length polymorphism analysis]. AB - OBJECTIVE: To establish a methed of cleavage fragment length polymorphism (CFLP) analysis with a primer labeled at the 5'-end with digoxigenin for genotyping of Chlamydia trachomatis (Ct). The methods for detection of Ct by major outer membrane protein (MOMP) gene (ompl) with nested polymerase chain reaction (ompl nPCR) were studied. The incidence of Ct infection in pregnant women, the common genotypes and vertical transmission rate of Ct in Chongqing area during the past one year was also investigated. METHODS: The samples were taken from cervical scrapes of parturient women and nasopharygeal swabs of their neonates from April 2003 to Feb. 2004 in Chongqing Women and Children's Health Care Institute. Totally 300 pairs (605 specimens) were detected by using ompl-nPCR, ompl-PCR (inside pair of primers was used directly) and plasmid-PCR. The results were judged by the modified gold standard (MGS). The ompl-nPCR amplified DNA was purified by recovery of DNA from agarose gel electroelution into dialysis bags. The DNA amplified from ompl-nPCR was sequenced by ABI PRISM 377 DNA sequencer. CFLP assay with a primer labeled at the 5'-end with digoxigenin was created for genotyping of Ct, and was primarily applied. RESULTS: The minimum detectable levels of ompl-nPCR and ompl-PCR corresponded to 2.5 elementary body (EB) and 25 EB, respectively. The sensitivity of ompl-nPCR was 10 times that of ompl-PCR. The positive rate of Ct in the samples from the pregnant women was 11% (33/300). The vertical transmission rate of Ct from mothers to their infants was 24.2% (8/33). The rate of Ct isolated from nasopharyngeal swabs 5 - 10 days after birth was 38.9% (7/18), which was significantly greater than that [3.0% (1/33)] detected within 24 hours after birth (chi(c)(2) = 8.79, P < 0.01). Of the 33 Ct-positive samples from pregnant women, 9 had vaginal delivery and 24 had caesarean section. The vertical transmission rates in vaginal delivery group and caesarean section group were 66.7% (6/9) and 8.3% (2/24), respectively (chi(c)(2) = 9.16, P < 0.01). Incidence of premature rupture of membrane in Ct-positive group was 30.3% (10/33), which was greater than that of Ct-negative groups (13.5%, 36/267, chi(2) = 6.40, P < 0.05). Four different patterns were observed in the 16 Ct-positive samples from 8 pregnant women and 8 matched maternal-infants by using CFLP, which were confirmed by DNA sequencing later. They were type E (3 pairs), type F (2 pairs), type H (2 pairs) and type D (1 pair). Each pair of matched maternal infantile samples presented identical CFLP pattern. CONCLUSIONS: This study revealed the infection rate of Ct in pregnant women, vertical transmission rate of Ct and the common genotypes of Ct in Chongqing Women and Children's Health Care Institute. The CFLP assay by using a primer labeled at the 5'-end with digoxigenin was first used for genotyping of Ct. The assay showed a good sensitivity and reproducibility, no radioactive contamination, and is simple. Therefore the assay is a potential new method for Ct genotyping. PMID- 15796797 TI - [Trend of the CD23+ B cells in children with infectious mononucleosis caused by Epstein-Barr virus]. AB - OBJECTIVE: Epstein-Barr virus (EBV) is a common causative agent of infectious mononucleosis (IM) and capable of efficiently immortalizing primary B cells into continuously growing lymphoblastoid cells in vitro. As B cell activation antigen, CD23 expression is induced by EBV infection of B cells and remains constitutively expressed at high levels in virtually all EBV-immortalized cells, which have been strongly linked to the development of B-cell lymphoproliferative disease and lymphoma. Whereas previous studies were performed in vivo in animals or ex vivo cultures, the present study aimed to explore the role of EBV-immortalized cells (CD23(+)/CD19(+)) in vivo analysis of children with EBV-IM. METHODS: In a prospective trial, a group of 30 patients with IM (18 boys and 12 girls) with mean age of 3.9 +/- 1.3 years (range 6 months to 8 years) were enrolled. Clinical diagnosis of IM was confirmed based on fever, lymphadenopathy, splenomegaly, lymphocytosis (> 50%), atypical lymphocytes (> 10%) in blood smears and the elevated levels of IgM antibody against EBV capsid antigen. The day of onset of fever was recognized as day 1 of illness. Blood samples taken during acute (3 - 5 days), early convalescent (about 11 - 15 days) and convalescent phase (about 30 - 45 days) were analyzed for expressions of CD19(+)/CD23(+), CD23, CD19 on peripheral blood mononuclear cells by flow cytometry (FCM) and was compared with those of control group. RESULTS: (1) The levels of CD23(+)/CD19(+) and CD23 expressions were markedly decreased in acute stage [CD23(+)/CD19(+) (2.22 +/- 1.47)%, (132 +/- 91)/mm(3); CD23 (3.12 +/- 1.88)%, (195 +/- 102)/mm(3)] and in early convalescent stage [CD23(+)/CD19(+) (4.51 +/- 2.25)%, (166 +/- 85)/mm(3); CD23 (5.55 +/- 2.76)%, (231 +/- 130)/mm(3)] in patients with IM as compared with those of the healthy controls [CD23(+)/CD19(+) (6.71 +/- 2.25)%, (215 +/- 68)/mm(3); CD23 (7.85 +/- 3.09)%, (249 +/- 86)/mm(3), respectively]. The earlier the history was, the lower the expressive levels were. The levels of CD23(+)/CD19(+) expressions returned to, but those of CD23 expressions exceeded, normal level in convalescent stage [CD23(+)/CD19(+) (6.72 +/- 2.16)%, (213 +/- 108)/mm(3); CD23 (9.46 +/- 2.73)%, (366 +/- 200)/mm(3)]. (2) There was a positive correlation in the expressions of CD23(+)/CD19(+) and CD23 among the three stages (P < 0.01). The positive correlation between the expressions of CD23(+)/CD19(+) and CD19 only occurred during acute stage (P < 0.01). There was no correlation between the expressions of CD23 and CD19 (P > 0.05). CONCLUSION: EBV-immortalized cells and CD23(+) cells were inhibited effectively during the acute and early convalescent stage of IM. With the recovery of the disease, they gradually recovered and the levels of CD23 expressions exceeded normal level in convalescent stage. PMID- 15796799 TI - [Present status of studies on rhinovirus]. PMID- 15796798 TI - [Application of flow cytometry to detect PP65 antigenemia for diagnosis and monitoring of human cytomegalovirus infection]. AB - OBJECTIVES: To evaluate the clinical significance of flow cytometry (FCM) to detect the cytomegalovirus (CMV) PP65 antigen in patients with CMV infection. METHODS: Samples from 35 patients without CMV infection were used as negative control. The definite diagnosis of CMV infection was based on the national criteria for CMV infection. All 136 patients with CMV infection were examined with the FCM to detect CMV PP65 antigen, real-time fluorescence quantitative polymerase chain reaction assay (RFQ-PCR) to detect CMV-DNA and ELISA to measure the serum level of IgM antibody against CMV. The results of these 3 assays in 2 groups (isolated organ involvement and disseminated diseases) were compared and the significance of PP65 antigenemia was evaluated. A short-term follow-up was undertaken in 18 patients. RESULTS: The percentages of PP65 positivity in blood mononuclear cells (MNC) and polymorphic nuclear leukocyte (PMNL) from 35 negative control patients were 0.21% +/- 0.09% with a range of 0 - 0.41% and 0.24% +/- 0.10% with a range of 0.12% - 0.48%, respectively, which were not significantly different (t = 0.425, P > 0.05). The 95(th) percentiles (P(95)) of PP65 in MNC and PMNL were 0.39% and 0.45%, respectively, so a cutoff value of >/= 0.50% was set. Of the 136 patients with CMV infection, 118 samples from 118 patients were positive for PP65 antigenemia with a positive rate of 86.8%, which was not statistically different from that (90.4%, chi(2) = 0.91, P > 0.05) of CMV-DNA detected by RFQ-PCR assay but it was significantly higher than that (45.6%, chi(2) = 51.50, P < 0.005) of the detection by IgM measurement. PP65 detection was correlated with urine CMV DNA amplification (chi(2) = 63.78, P < 0.01) while the different detection rates between the two assays were not statistically significant (chi(m)(2) = 1.78,P > 0.05). PP65 detection was not correlated with serum IgM measurement while the detection rates between the two were significantly different (chi(m)(2) = 52.92,P < 0.01). No significant difference was found between the detection rates of CMV infection in MNC (45/53, 84.9%) and PMNL (43/53, 81.1%) (chi(m)(2) = 0.25, P > 0.05). Higher PP65 antigenemia level was correlated with systemic CMV infection, while lower level of PP65 was either in the patients with isolated organ involvement by CMV (chi(2) = 38.51, P < 0.005) or less severe in patient's situation. PP65 antigenemia of CMV infection returned to lower level or negative in recovery stage and increased when condition of patients deteriorated. CONCLUSIONS: PP65 antigenemia detection by FCM is effective in the diagnosis of the active CMV infection. Quantitative monitoring of PP65 antigenemia is useful in the evaluation of patients with CMV infection. PMID- 15796800 TI - [Progresses in studies on adult stem cells]. PMID- 15796802 TI - [A case of brucellosis with cytomegalovirus infection]. PMID- 15796801 TI - [Summary of the symposium on immunity, infection and antibiotic resistance]. PMID- 15796803 TI - [Association between dopamine beta hydroxylase gene and attention deficit hyperactivity disorder complicated with disruptive behavior disorder]. AB - OBJECTIVE: Attention deficit hyperactivity disorder (ADHD), a common behavior disorder of childhood, is a highly heterogeneous disease frequently accompanied by various mental disorders, including disruptive behavior disorder (DBD). Studies show that children suffering from ADHD with DBD are at higher risk of antisocial personality, substance abuse, and social adaptations disorder at their adulthood. The dopamine beta hydroxylase (DbetaH) is the key enzyme to ADHD since it catalyzes the conversion of dopamine to norepinephrine, and dysfunction there of is believed to be one of the causes of the disorder. To explore the association between DBH gene and ADHD complicated with or without DBD, the authors analyzed the transmission of a novel polymorphism DBH -1021C-->T, which is found associated with plasma DbetaH activity, in ADHD nuclear families using transmission disequilibrium test (TDT). METHODS: Consensus diagnoses were based on the DSM-IV. The samples included those from 292 Chinese Han nuclear families with ADHD probands. Genotypes of DBH -1021C-->T polymorphism were determined by PCR amplification, endonuclease digesting and electrophoresis. The transmission of DBH -1021C-->T polymorphism in ADHD nuclear families with or without DBD was analyzed by TDT. RESULTS: The results showed that there was transmission disequilibrium between DBH-1021C-->T polymorphism and ADHD with or without DBD. In ADHD comorbid with DBD, T allele was preferentially transmitted (P < 0.05); and in ADHD without DBD, so was the C allele (P < 0.05). Among the three subtypes of ADHD, only ADHD-C subtype with DBD had an increased transmission of T allele (P < 0.05). CONCLUSION: There is an association between DBH gene and ADHD comorbid with or without DBD, but the preferential transmission alleles are different. The low activity T allele is increased to transmit in ADHD with DBD, while the high activity C allele is preferentially transmitted in ADHD without DBD. The results support the proposition that the genetic mechanism is different between ADHD comorbid with or without DBD. We also found that only ADHD-C subtype with DBD is associated with DBH -1021C-->T polymorphism in three subtypes of ADHD, which may suggest that there is a more intense relationship between DBD and ADHD-C subtype. PMID- 15796805 TI - [Hereditary coagulation factor VII deficiency in a case]. PMID- 15796804 TI - [Diagnosis of thalassemia by using genechips]. AB - OBJECTIVE: The new technology of genechip is exerting a significant impact on the identification of thalassemia, and allows to rapidly and efficiently detect a number of molecular disorders. The aim of the present study was to explore the application value of the diagnostic genechip in determining thalassemia. METHODS: The subjects group consisted of 62 children with alpha thalassemia and 93 children with beta thalassemia (60 with thalassemia trait, 33 with thalassemia major) from Guangdong province were tested from July 2002 to July 2003; 115 were males and 40 were females, the age ranged from 1 day to 11 years. These children had mild, moderate or severe anemia. Laboratory examinations showed microcytosis and hypochromocytosis. DNA was extracted from ACD coagulated blood with Invisorb DNA extraction kit. After preparation, the alpha and beta globin gene organization and structure of sample was analyzed by genechip technology. RESULTS: (1) Heterozygotes, homozygotes and compound heterozygotes of the three commonest deletional alpha thalassemia (--(SEA), -alpha(3.7), -alpha(4.2)) and two non-deletional alpha thalassemia (HbCS, HbQS) were successfully detected. (2) Eight different mutations were identified in 126 chromosomes among 60 cases with beta thalassemia traits and 33 cases with beta thalassemia major, namely CD41-42, IVS-II-654, TATAbox-28, CD17, CD71-72 (+A), betaE(26) CD27-28 (+C) and CD71-72 (+T), with the frequencies of 33.5%, 23.9%, 15.0%, 15.0%, 4.8%, 2.4%, 2.4% and 1.5% respectively. (3) Eight cases (13.3%) in 60 thalassemia traits, two cases (6%) 33 cases with thalassemia major were detected to be combined with alpha thalassemia mutation. CONCLUSION: (1) The usage of genechip in identifying thalassemia mutations has the advantages of simplicity, economy and shorter time. This technique does not use radioisotope and could also detect alpha and beta thalassemia mutations simultaneously. (2) The occurrence of alpha and beta thalassemia dual heterozygotes is frequent in Guangdong province and the genechip technology is important in genetic counseling and prenatal diagnosis of thalassemia in this area. PMID- 15796806 TI - [Complications of transcatheter interventional occlusion of ventricular septal defects]. AB - OBJECTIVE: To investigate the complications and their prevention in patients with perimembranous ventricular septal defect (VSD) experiencing transcatheter interventional occlusion. METHODS: From July, 2002 to May, 2004, totally 262 (138 males, 124 females) perimembranous VSD patients underwent transcatheter interventional occlusion in the department. The age ranged from 2 to 18 years (mean 9.3 +/- 5.8 years), and the body weight ranged from 11.0 to 65.0 kg (mean 30.5 kg). The implanted occluder with imported Amplatzer eccentric perimembranous VSD occluder, domestic double-disk, domestic eccentric perimembranous VSD and PDA occluder were attempted in 12, 212, 28 and 6 cases, respectively. Among them, 3 cases were complicated with ductus arteriosus, 4 with atrial septal defects and 1 with patent ductus arteriosus plus atrial septal defect. The associated defects were treated with other occluders at the same time. RESULTS: The diameters of VSD measured by transthoracic echocardiography before occlusion and by ventriculography during the procedure ranged from 2.7 to 13 mm (mean 6.5 mm) and from 1.3 to 14.0 mm (mean 6.8 mm), respectively. The defects in 256 cases (97.7%) were successfully occluded. The implant occluder was from 4 to 16 mm in size. Serious complications occurred in 8 cases (3.1%), including 5 cases of high degree atrioventricular block (AVB) (2.0%), 2 cases of hemolysis (0.8%) and 1 case of displacement of the occluder (0.4%). Amongst 5 cases of high degree AVB, permanent pacemaker was implanted only in 1 case due to high degree AVB 50 days after treatment, whereas the others all recovered. Two cases of hemolysis were completely recovered. One case of occluder displacement was emergently transferred to cardiac surgery. Other complications included residual shunt during 6 month follow-up in 1 case (0.4%), minor aortic regurgitation in 5 cases (2.0%), minor tricuspid regurgitation during 6 month follow-up in 4 cases (1.6%), and accelerated idioventricular rhythm or accelerated atrio-ventricular junctional tachycardia combined with atrioventricular interference-dissociation by ECG in 75 cases (29%) but they recovered after 3 - 5 days of corticosteroid treatment. Forty-one cases (16%) complicated with complete or incomplete right bundle branch block, of whom 50% recovered during follow-up. Still, there were 4 cases (1.6%) who were complicated with complete left bundle branch block but were all recovered during follow-up and 26 cases (10%) who were complicated with intraventricular block. There were no Significant difference in complications between cases who accepted imported Amplatzer occluders and domestic occluders. CONCLUSION: Transcatheter interventional occlusion is a safe, effective and ideal method with low incidence of serious complications for perimembranous VSD. Long term follow-up and clinical research should be carried out to improve the level of VSD interventional therapy. PMID- 15796807 TI - [Expression of tumor necrosis factor alpha mRNA in adipose cell of intrauterine growth retarded rats and its relation to insulin resistance]. AB - OBJECTIVE: To investigate the association between the expression of turnor necrosis factor alpha (TNF-alpha) mRNA in fat tissue of intrauterine growth retarded (IUGR) rats and insulin resistance, and the long-term effects of early different nutritional diet. METHODS: The IUGR rat model was established by food restriction of pregnant rats. A total of 32 newborn IUGR rats were randomly divided into 4 groups: IUGR model (S/N) group, IUGR high caloric diet (A) group, IUGR high caloric and high protein diet (B) group, IUGR high protein diet (C) group. Only the mother rats were given those different diets individually, and all IUGR newborn pups were lactated for 3 weeks. From the beginning of the 4(th) week, all IUGR pups were weaned and fed with normal diet till the end of the experiment. Eight normal birth weight newborn rats were used as the control group fed with the normal diet. Weight, perirenal fat weight, fasting glucose and insulin concentration and quantified TNF-alpha mRNA expression in adipose cell were measured at the 48(th) week. The insulin sensitive index (ISI) and the relation index between TNF-alpha mRNA and fat weight, fat weight/body weight (fw/bw) ratio and ISI were calculated. RESULTS: ISI of IUGR model group, IUGR A and B groups was lower than normal control group, while perirenal fat weight, fw/bw and the expression of TNF-alpha mRNA in adipose cells were all significantly higher (P < 0.05 or 0.01). There were no significant differences in these indexes between IUGR C group and normal control groups (P > 0.05). A positive correlation was found between TNF-alpha mRNA and fat weight and fw/bw (r(1) = 0.755, r(2) = 0.782, P = 0.000). Significant inverse associations between ISI and TNF-alpha mRNA (r = -0.556, P = 0.000) and fw/bw (r = -0.513, P = 0.02) were also found. CONCLUSION: The occurrence of insulin resistance in IUGR rats is possibly associated with central obesity and accumulation of the abdominal fat and adipose cell over-expression of TNF-alpha. The adipose TNF-alpha may be an important pathogenic factor of insulin resistance of IUGR. High protein diet is a reasonable nutritional intervention. Because it promotes the skeleton muscle catch-up growth but not fat catch-up growth, it can avoid the occurrence of central obesity and insulin resistance in IUGR rats. PMID- 15796808 TI - [Pulmonary cryptococcosis accompanied by eosinophilia in a case]. PMID- 15796809 TI - [Cause and mortality analysis of acute intracranial hypertension and cerebral edema in pediatric intensive care unit]. AB - OBJECTIVE: Acute intracranial hypertension/cerebral edema (ICH/CE) is an increase in brain volume caused by an absolute increase in cerebral tissue water content. Severe ICH/CE is often associated with a higher mortality and higher neurological consequence rate in intensive care unit. However, little relevant information is available on critical condition of central nervous system in children. The aim of this survey was to study the causes, clinical epidemiology and risk factors of critical illness with ICH/CE in pediatric intensive care unit (PICU). METHODS: Case records of critically ill patients with ICH/CE admitted to PICU in Children's Hospital Affiliated to Shanghai Jiaotong University during the period from January, 1999 to December, 2003 were reviewed for causes, case fatality rate, prognosis and relationship with multiple organ dysfunction syndrome (MODS). Univariate analyses were performed to identify risk factors associated with ICH/CE. RESULTS: During the 5 years, 1446 cases with critical illnesses were admitted and ICH/CE developed in 216 patients. The leading causes of ICH/CE were central nervous system infection (27.8%), accidental injuries (22.4%), and sepsis (10.2%). The overall mortality of the patients with ICH/CE was 29.2%. The mortality showed no significant change during the years from 1999 to 2003 (chi(2) = 0.371, P = 0.985). There was no significant difference in mortality of patients with ICH/CE between those with and without neurological diseases (chi(2) = 0.546, P = 0.460). Univariate analyses involving 12 factors indicated the following risk factors: younger age, number of failed organ, lower pediatric critical illness score, underlying diseases, abnormal respiration and change in size of pupil (P < 0.05 or < 0.001). The following factors were not associated with higher risk of death from ICH/CE: sex, organ of primary disease, Glasgow coma score ( 7) on admission, elevated blood pressure and anterior fontanelle change (P > 0.05). CONCLUSIONS: The mortality of ICH/CE remains high since 1999. Central nervous system infection, accidental injuries, and sepsis were leading causes of ICH/CE in PICU of the hospital. Children who had ICH/CE due to younger age, lower pediatric critical illness score, and complicated with MODS had a higher mortality rate. PMID- 15796810 TI - [Effects of hypothyroidism on apoptosis and the expression of Bcl-2 and Bax gene in the neonatal rat hippocampus neurons]. AB - OBJECTIVE: During the critical period of brain development, insufficiency of thyroid hormone results in severe mental retardation and learning deficit. This study was designed to investigate the effects of hypothyroidism on apoptosis and the expression of Bcl-2 and Bax gene in the developing rat hippocampus neurons and to explore the mechanism of brain development regulated by thyroid hormone. METHOD: Hypothyroidism was induced by administration of propylthiouracil (PTU, 50 mg/d) solution to the dams from gestational day 15 by gavage. Pups from both hypothyroid and control groups were harvested at postnatal day 1 (P1), P5, P10 and P15, respectively. Blood samples were collected at the time of death for the determination of thyroid hormone. Serum free tri-iodothyronine (FT(3)) and free thyroxine (FT(4)) were measured by using chemoluminescence. Hippocampus collected from the control and hypothyroid pups were examined under light and transmissional electron microscopy. Measurement of DNA fragmentation was carried out by agarose gel electrophoresis. The expression of Bcl-2 and Bax protein in the developing rat hippocampus neurons was performed by Western blotting. RESULTS: Significantly lower circulating FT(4) and FT(3) levels confirmed the hypothyroid status of the experimental pups. The shrunken and contracted degenerations increased in hippocampus neurons of hypothyroid pups under light microscopy. Enhanced apoptotic cells were found in hippocampus neurons of hypothyroid pups under transmission electron microscopy, especially at P10 and P15. Extensive DNA fragmentation was seen throughout development in hippocampus of hypothyroid pups, but not in the euthyroid controls except for basal level at P10. The expression of Bcl-2 in the hippocampus neurons of hypothyroid pups was significantly lower than that of euthyroid controls at all stages of development (P1: 1.95 +/- 0.27 vs. 2.59 +/- 0.19, P < 0.05, P5: 1.86 +/- 0.24 vs. 2.47 +/- 0.17, P < 0.05, P10: 1.29 +/- 0.22 vs. 1.86 +/- 0.28, P < 0.05 and P15: 1.21 +/- 0.27 vs. 2.18 +/- 0.17, P < 0.01, respectively). The relative amount of expression varied significantly with age in the control pups. The level of Bcl-2 was high in hippocampus neurons of euthyroid at P1, P5, and decreased significantly at P10, and showed a trend of recovery at P15. Similar age-related variation in the expression of Bcl-2 gene was observed in the hypothyroid group at P1, P5 and P10, but the level was maintained low at P15. The expression of Bax in the hippocampus neurons of hypothyroid pups was significantly higher than that of control pups at all stages of development (P1: 1.69 +/- 0.14 vs. 1.24 +/- 0.23, P < 0.05, P5: 1.78 +/- 0.16 vs. 1.29 +/- 0.17, P < 0.05, P10: 1.92 +/- 0.18 vs. 1.45 +/- 0.14, P < 0.05 and P15: 1.86 +/- 0.14 vs. 1.51 +/- 0.12, P < 0.05, respectively). The ratio of Bcl-2/Bax in hippocampus neurons of hypothyroid pups was lower than that of age-matched controls (P1: 1.16 +/- 0.17 vs. 2.12 +/- 0.35, P < 0.05, P5: 1.05 +/- 0.16 vs. 1.94 +/- 0.36, P < 0.05, P10: 0.68 +/- 0.17 vs. 1.29 +/- 0.16, P < 0.05 and P15: 0.67 +/- 0.19 vs. 1.45 +/- 0.22, P < 0.01, respectively). CONCLUSION: Thyroid hormone significantly prevents apoptosis of hippocampus neurons. Congenital hypothyroidism increases not only the extent but also the duration of apoptosis by down-regulation of the anti-apoptotic gene Bcl 2 and maintaining a high level of the pro-apoptotic gene Bax. PMID- 15796811 TI - [Rat bone mesenchymal stem cells in vitro differentiation into neuron-like cells induced by Salvia miltorrhiza for injection]. PMID- 15796812 TI - [Effect of losartan on heme oxygenase-1 expression in volume overloaded rats]. PMID- 15796813 TI - [Effects of high concentration of oxygen on heme oxygenase-1 and carbon monoxide in the lung of neonatal rats]. PMID- 15796814 TI - [Relationship between tumor necrosis factor-alpha promoter G238A and G308A polymorphism and susceptibility to hepatitis B virus intrauterine infection]. PMID- 15796816 TI - [Invasive mucormycosis in neonates, analysis of 8 cases]. PMID- 15796815 TI - [Idiopathic CD4+ T-lymphocytopenia in a child with disseminated cryptococcosis]. PMID- 15796818 TI - [Effects of combined artemether and dihydroarteamisinin in treatment of children with falciparum malaria]. PMID- 15796817 TI - [Effect and safety of famotidine for therapy of gastrointestinal bleeding in children]. PMID- 15796819 TI - [Analysis of 150 children cases with multiple organ dysfunction syndrome]. PMID- 15796820 TI - [Report of 3 cases with ataxia-telangiectasia in a family]. PMID- 15796821 TI - [Report of a survived neonate born to the mother who had liver transplantation]. PMID- 15796822 TI - [Congenital chloride diarrhea in a case]. PMID- 15796823 TI - [Systemic lupus erythematosus with Evan's syndrome as primary symptom in a case]. PMID- 15796824 TI - [A case with severe infestation with Armilliferiasis moniliformis]. PMID- 15796826 TI - [Pursue one's study and cultivate one's morality in scientific research]. PMID- 15796825 TI - [Summary of the 13th international congress of pediatric nephrology]. PMID- 15796829 TI - [Prospect of the application of bio-medical engineering technology in wound repair]. PMID- 15796827 TI - [Retrospect and prospect]. PMID- 15796831 TI - [Relationship between the management and prognosis of deep partial thickness burns]. PMID- 15796833 TI - [Improving the management of deep partial thickness burn wound]. PMID- 15796835 TI - [Experimental study of autologous skin grafting on retained denatured dermis for the treatment of partial thickness burn wound]. AB - OBJECTIVE: To provide experimental evidence for the feasibility of repair of deep partial thickness burn wounds with autologous skin grafting on retained denatured dermis. METHODS: Deep partial thickness burn wounds 3.5 cm x 3.5 cm in size were produced on the back of SD rats. Superficial tangential excision was performed on the burn wounds on 2 - 5 postburn days with the preservation of denatured dermis. Split thickness autoskin was grafted on the wounds immediately. Tissue samples of whole layer of the skin were harvested from the grafted sites at different time points after the skin grafting, with 8 rats in each group. The morphology and the changes in the collagen fibres in the above tissue samples were observed by light microscopy (LM) and the changes in the biodynamics were determined. The normal skin from the same rat receiving skin grafting was employed as the control. RESULTS: (1) Vitreous degeneration was observed in the preserved denatured dermis. (2) The grafted skin was fused to the burn wound on the 7th day after autoskin grafting in rats. The dermal papillae and reticular layer could be discernible under LM. The thickness, structure and morphology of the skin on the grafted area were similar to normal tissue on the 21st day after grafting with atrophic hair follicles. The density of collagen fiber cord increased gradually, and almost fused at last. (3) The tensile strength, and maximum strain value of the rat skin after operation increased gradually, approaching normal on 60 post operational day. CONCLUSION: The denatured dermis can recover gradually with normal structure and morphology after the application of autoskin on it for the repair of deep partial thickness burn wounds. PMID- 15796836 TI - [Preliminary study on the improvement of wound microcirculation and retrospection on several methods of the management of deep partial thickness burn wound]. AB - OBJECTIVE: To analyze several methods of wound repair for deep partial thickness burn wounds retrospectively, so as to evaluate the significance of improvement of wound microcirculation on wound healing. METHODS: (1) 2,976 burn patients admitted to our department were enrolled in the study, among them 614 undertook tangential excision, 32, eschar abrasion, 86 allo-skin coverage after debridement, 1836 tropical application of silver sulfadiazine and 408 with traditional Chinese medicine (Jing Wan Hong ointment) with gauze bandage. The results of the management with different methods were compared. (2) Rat model with deep partial thickness burn was reproduced and topical application of silver sulfadiazine was given. The rats were randomly divided into control (n = 10, with normal saline injected via caudal vein within 5 minutes postburn), and treatment (n = 10, with batroxobin injected via caudal vein within 5 minutes postburn) groups. The blood flow perfusion unit in the wound skin was measured before burn and at 0.5 to 72 postburn hours by Laser Doppler. The wound healing rate, contraction rate and wound healing time in each group were calculated on 14 and 18 postburn days (PBDs). The number of hair follicles after wound healing was observed by histological method. RESULTS: (1) The burn wound treated by tangential excision healed within 2 to 3 post operation weeks (POWs), with the healing rate of 94.8% in patients with burn covering 50% - 70% TBSA and 93.4% in those with burn of 80% approximately 98% TBSA. The healing time of patients with allo-grafts coverage after eschar abrasion was 13.8 +/- 2.1 days without scar formation. The wound healing time was 18.0 +/- 2.3 day in 82 patients with allo graft coverage after debridement, and it was 26.0 +/- 3.2 days with subeschar healing in 1658 patients with topical application of silver sulfadiazine. Infection in burn wound was encountered in most patients undergoing traditional Chinese medicine bandage treatment with wound healing time of 26.0 +/- 2.8 days in the lower extremities. (2) The blood flow perfusion unit of the rats in the treatment group was significantly higher than that in the control group (P < 0.01). The wound healing rate in treatment group on 14 and 18 PBD was obviously higher than that in the control group (P < 0.01). But the wound contraction rate in the two groups was similar (P > 0.05). The wound healing time in treatment group was much shorter than that in control group (P < 0.01). A few hair follicles remained in the dermis of the rats in the control group on 30 PBD, and the number was evidently smaller than that in the treatment group (P < 0.01). CONCLUSION: Early tangential excision and eschar abrasion remained better methods in the management of deep partial thickness burn wounds, as they could ameliorate burn wound infection, shorten treatment period, raise wound healing rate and quality. Application of batroxobin could accelerate wound healing rate by improving wound microcirculation in deep partial thickness burn wound. PMID- 15796837 TI - [Observation of the effect of the mixed composite skin graft on deep partial thickness burn wounds]. AB - OBJECTIVE: To evaluate the effect of mixed composite skin graft on the deep partial thickness burn wounds after tangential excision in burn patients. METHODS: Tangential excision was performed in 30 extremities of 23 burn patients within 3 postburn days (PBDs). Then large pieces of homologous acellular dermal matrix were grafted onto the superficial fascia with razor thin autoskin on top of them. The survival rate of skin grafts, the appearance and the functional recovery of the extremities were observed on 10 to 12 post operative day (POD). Skin samples from a healed wound of a patient were harvested three months after the injury for pathologic examination. RESULTS: The survival rate of the composite skin grafts was 93%. Necrosis was encountered in 7% of the grafts in the lower extremities due to the poor fixation of the grafts leading to separation of autologous skin and the dermal template, and also due to infection resulting in lysis of the grafts. The grafted skin was excellent in the appearance and elasticity, and function of the injured extremities recovered well after grafting after 3 - 6 months of follow-up. Epidermal and dermal texture was also good as shown by pathologic examination. CONCLUSION: Mixed composite skin grafting after early tangential excision might be an ideal and effective method in the management of deep partial thickness burn wounds. PMID- 15796839 TI - [Influence of tangential excision within 24 postburn hours on the local wound inflammatory response in patients with deep partial thickness burn]. AB - OBJECTIVE: To investigate the influence of early tangential excision within 24 postburn hours on local wound inflammatory response and tissue injury in patients with deep partial thickness burn. METHODS: Twelve patients with deep partial thickness burn were enrolled in the study. Tangential excision was performed within 24 postburn hours. The same wound of a patient was divided into three areas for harvesting tissue samples, i.e. pre-operation, post-operation and non operation areas. For each patient, the biopsies from the pre-operation area were harvested before tangential excision, while those from post-operation and non operation areas were harvested during 5 to 7 postburn days (PBDs). The contents of IL-8, MPO and MDA in wound tissue were determined by tissue culture and chemical colorimetry, respectively. HE and Mason's staining were employed to assess the degree of necrosis of the wound tissue. RESULTS: The IL-8, MPO and MDA contents in the local wound tissue after tangential excision were 6.83 +/- 1.85 microg/L, 4.07 +/- 0.87 U/g, and 8.94 +/- 5.66 micromol/g, respectively, which were significantly lower than those from non-operation area (P < 0.01). The inflammatory response in the non-operative wound area was distinct with expansion of necrotic tissue area. In contrast, the local inflammatory response of the wound after tangential excision was ameliorated without the enlargement of necrotic tissue area. CONCLUSION: Tangential excision within 24 postburn hours could be beneficial in ameliorating local inflammatory response, and in preventing progressive deepening of the burn wounds, thus it could accelerate wound healing in patients with deep partial thickness burn. PMID- 15796840 TI - [Long-term result of repair of deeply burned hands with large sheet of split thickness autoskin grafting with the preservation of denatured dermis]. AB - OBJECTIVE: To evaluate the long-term results of repair of burn hands with large sheet of split-thickness autoskin grafting with the preservation of denatured dermis. METHODS: One hundred and fifty-two hands in 86 burn patients with deep partial thickness and full thickness burn were enrolled in the study. The burned hands were treated by tangential excision and grafted with large sheet of split thickness autoskin with the preservation of denatured dermis. The patients were followed-up from 3 months to 3 years. The skin color, elasticity, degree of contracture and the functional grading of the operated hands were observed. RESULTS: Good function was found in one hundred and forty-one out of the 152 burn hands (92.8%). For the rest 11 hands, pigmentation was found in 4, poor appearance in 4, and 3 hands with both poor appearance and function. CONCLUSION: Large sheet of split-thickness autoskin grafting with the preservation of denatured dermis could be an optimal choice for the management of hands with deep partial thickness burn, and it could restore the appearance and function of the hands satisfactorily. PMID- 15796841 TI - [Influence of human telomerase reverse transcriptase gene transfection on the proliferation of human embryonic fibroblasts]. AB - OBJECTIVE: To investigate the influence of human telomerase reverse transcriptase (hTERT) gene transfection on the proliferation of human embryonic fibroblasts (hEF). METHODS: hEFs were cultured in vitro. Sense recombinant eukaryotic plasmid (pIRES2-EGFP-hTERT) and pIRES2-EGFP vacant vector were transfected into hEF respectively with Lipofectin reagent, and were named as hEF-hTERT and hEF-EGFP. The hTERT, Id1, PCNA and I, III type collagen expression in these cells were detected by Western blot. Then the cell cycle and growth curve were measured and plotted with flow cytometry and MTT method, respectively. RESULTS: 1. The expression of hTERT, Id1, PCNA, type I and III collagen in hEF-hTERT were much higher than that in hEF and hEF-EGFP. 2. As shown in the growth curve, the OD value of hEF-hTERT at 4 to 6 days after culture was obviously higher than that of hEF and hEF-EGFP (P < 0.05), while no difference existed between hEF and hEF-EGFP from 1 to 6 days after culture (P > 0.05). 3. The cell number in G0/G1 phase in hEF-hTERT was less than that in hEF and hEF-EGFP. The cell number of hEF-hTERT in S and G2/M phase and its proliferation index (57.47%) increased when compared with that in hEF-EGFP (13.13%) and hEF (17.38%), but there was no difference between hEF and hEF-EGFP. CONCLUSION: Exogenous hTERT gene transfection could promote the proliferative capacity of hEF. PMID- 15796842 TI - [Observation of the grafting of platelet-derived growth factor gene-modified artificial composite skin on rat wounds]. AB - OBJECTIVE: To observe the effects of the grafting of a platelet-derived growth factor gene-modified artificial composite skin on rat wounds with full thickness defect. METHODS: Platelet derived growth factor-B (PDGF-B) eukaryotic expression plasmid was constructed, and the fibroblasts were transfected with it by liposome mediation. Artificial composite skins 1 and 2 were constructed respectively. The skin1 was composed of keratinocyte, porcine acellular dermal matrix and PDGF-B gene-transfected fibroblasts while the skin 2 contained keratinocyte, porcine acellular dermal matrix and fibroblasts. The two kinds of composite skin were grafted onto wounds on the rat back to form composite skin group 1 (C1) and 2 (C2), respectively, with 18 rats in each group. Eight rats with wounds without treatment served as control (C) group. The survival rate of the composite skin was observed at 2 post-operative weeks (POWs). The rat wounds were examined grossly on 2, 4 and 6 POWs for the calculation of wound contraction rate. Wound tissue samples were harvested for histological examination. RESULTS: (1) Up to 2 POWs, 14 grafts in C1 group survived completely, 3 with partial survival and 1 failure. In C2 group, 10 skin grafts survived completely, 4 with partial survival and 4 failures. (2) A scab was formed in the wound at 2 POW in C group. The surface of the grafted skin in C1 group was smooth, elastic, and showed good anti friction properly, and it was better in quality compared with that in other two groups at 6 POW. (3) The wound contraction rate of the grafts in C group of rats was higher than that in C1 and C2 groups at 2, 4 and 6 POWs, while that in C1 was lower than that in C2 group. (4) Capillary formation was more intense in the grafted skins in C1 group at 2 POWs, and the epithelia differentiated well into 7 to 10 layers of epithelial cells with compact and orderly arrangement and evenly distributed fibrous tissue at 6 POWs. CONCLUSION: Repair of the wound with artificial composite skin containing PDGF-B gene could improve the quality of wound healing. PMID- 15796844 TI - [The histological changes and the revascularization process in the grafted dermal substitutes]. AB - OBJECTIVE: To observe the revascularization process and histological changes in the dermal substitutes after grafting. METHODS: Twenty-one SD rats were enrolled in the study and were randomly divided into swine acellular dermal matrix (sADM), human acellular dermal matrix (hADM), and artificial dermal equivalent (Integra) groups according to different dermal substitutes grafted underneath the skin of SD rats. The gross appearance of the grafts was observed, and the tissue biopsies were harvested at 2, 3, 4, 7, 10, 14, 21, 30, 60, 90, 120, 150, 180 post grafting day (PGD) for the observation of the revascularization process and their histological changes. RESULTS: Gross observation: The incision in each group healed well without local swelling and inflammatory response after grafting. The grafts had a compact contact with the wounds. The texture of the grafted dermis in each group was soft and could not be felt from the surface of the body on 90 PGD. The presence of the grafts could be discerned on 180 PGD in all the groups, but some of them became smaller in size and thinner, even hard to identify in a few. Fibroblasts, neutrophils and lymphocytes migrated into the grafts from 2PGD on. New capillary sprouts from the receiving beds could be observed after 3PGD. Affluent capillary nets formed in the grafts during 30 to 60 PGD. The structure of the grafts became nearly unidentifiable from the native dermis after 150 PGDs. Absorption and degradation to various degrees occurred in some of the grafts after 180 PGD. CONCLUSION: The revascularization of the three dermal substitutes could begin shortly after grafting. The sponge-like structure of the substitutes was advantageous for the migration of the host fibroblast into the substitute and for the secretion of the new extra-cellular matrix. The dermal substitutes could last in the wound for a long time with partial absorption and degeneration. PMID- 15796845 TI - [Comparison of various methods of early management of hydrofluoric acid burn in rabbits]. AB - OBJECTIVE: To compare the effects of different kind of methods in the management of hydrofluoric acid burn in early postburn stage in rabbits. METHODS: Thirty three rabbits were inflicted with burn by 55% of hydrofluoric acid covering 5% TBSA, and were randomly divided into 3 groups, i.e. A (n = 13, with 5 ml.kg( 1).h(-1)of isotonic saline intravenous infusion), B (n = 10, with isotonic saline and 50 g/L of calcium gluconate infusion in dose of 20 mg/kg at different time points), and C (n = 10, with the same treatment as B group, and with excision of burn wound at 0.5 post burn hour) groups. The serum levels of fluorine and calcium were determined before and after various postburn hours, and the mortality rate was statistically analyzed. RESULTS: (1) The serum level of fluorine in A (8.37 +/- 2.62 mg/L) and B (8.59 +/- 2.25 mg/L) groups reached the peak value at 1 postburn hour (PBH), which was 107 times higher than that before the burn injury. The serum level of fluorine in B group was significantly lower than that in A group at 24 PBH (P < 0.05), while that in C group declined to (6.20 +/- 0.23) mg/L, which was obviously lower than that in A and B groups (P < 0.01). (2) The serum calcium level declined after burns, reaching the lowest level at 8 to 12 PBH. and began to increase at 24 PBH. Compared with normal calcium value, the serum level of calcium in A, B and C groups declined to as much as 46%, 32% and 26%, respectively. Statistically significant difference was found between C and B groups (P < 0.01). (3) The mortality rate in the three groups within 72 PBH were 30.8%, 12.5% and 0.0%, respectively. CONCLUSION: Early removal of burn area and calcium supplementation could help quickly decrease blood fluorine, reverse the fatal hypocalcemia and the multiple systemic toxic injury in rabbits inflicted with hydrofluoric acid injury. PMID- 15796847 TI - [Clinical investigation in the expression of matrix metalloproteinase 2 mRNA in the hyperplastic scar in burn patients]. AB - OBJECTIVE: To investigate the dynamic expression of matrix metalloproteinase 2 in the hyperplastic scar in burn patients. METHODS: Twenty-three pieces of hyperplastic scar tissue samples were harvested from the patients with deep partial thickness burn wound during the early, middle and late periods after natural wound healing. In situ hybridization technique and positive cell percentage counting were employed to determine the expression and the distribution of MMP-2 mRNA in the scar tissue from different periods with that of the middle period as the control. RESULTS: MMP-2 mRNA was mainly expressed in the fibroblasts in scar tissue. The positive percentage of MMP-2 in early period was 13.9 +/- 3.6%, and it increased thereafter, till it reached 68.7 +/- 5.2% in the middle period. However, it declined to 2.0 +/- 0.9% in the late period. CONCLUSION: MMP-2 might be closely related to the process of scar tissue remodeling. PMID- 15796849 TI - [Effects of allogeneic hepatic nonparenchymal cell on the survival of grafted skin in mice]. AB - OBJECTIVE: To investigate the effect of allogene hepatic nonparenchymal cell (NPC) on the survival of grafted skin in mice and its underlying mechanism. METHODS: Sixty-five C(3)H and fifty-eight C(57)BL/6 mice were employed in the study. Twenty C(3)H mice were used as skin donor and forty as the source of hepatic NPC. The rest five served as the stimulators of mixed lymphocyte culture (MLC) before and on the 7th, 18th, 30th, and 60th day after NPC infusion with 1 at each time point. MLC was determined and expressed as count per minute (CPM). Fifty-eight C(57)BL/6 mice were further divided into experimental (E, n = 50) and control groups (C, n = 8). The mice in C group only underwent skin grafting without NPC infusion. The mice in E group received with 2 x 10(7) NPC via caudal vein, followed by peritoneal injection of cytoxan (200 mg/kg) 48 hours later; They were grafted with skin donated from C(3)H mice 18 days after injections. The survival time of the mice in the two groups was observed. The serum levels of interleukin-4, chimera and MLC in the two groups were determined before and on 7th, 18th, 30th, 60th days after NPC infusion, and micro-chimera were aslo assessed on the 1st and 3rd day after NPC infusion. Five mice were sacrificed at each time point. RESULTS: The survival time of skin graft in E group (70.0 +/- 17.2 day) was obviously longer than that in C group. The serum levels of IL-4, chimera in E group were increased gradually, while MLC response decreased gradually. The serum IL-4 level reached 251.5 +/- 11.0 ng/L and splenic chimera level to 26.30 +/- 1.04% on the 60th day after NPC infusion. CONCLUSION: The high levels of IL-4 and chimera might play important roles in inducing and maintaining immune tolerance. PMID- 15796851 TI - [A practical method for the calculation of wound water loss in burn patients]. AB - OBJECTIVE: To evaluate a calculating method of wound water loss (WWL) in hospitalized burn patients. METHODS: Eighty-five adult burn patients with total burn area over 50% were enrolled in the study. The WWL formula after shock stage (after 3rd postburn day) was given as: WWL (ml/%/kg/24 h) = [input (ml/24 h) urine output (ml/24 h)-500 ml] / actual burn wound area(%) / body weight (kg). The result was compared with that determined by other formulas such as Davies, S.M.S.P and Lamke and palm rule. RESULTS: The calculated daily WWL in burn patients after shock stage was found out to be 0.9 +/- 0.1 ml/%/kg/24 h by this method. This figure is comparable with that calculated by several different methods. Palm rule could be simply applied in the patients with body weight around 60 kg. CONCLUSION: This method and palm rule were proved to be simple and practical for the calculation of WWL in burn patients, which could be applied in clinical work. PMID- 15796853 TI - [The influence of dermal template on the expressions of signal transduction protein Smad 3 and transforming growth factor beta1 and its receptor during wound healing process in patients with deep burns]. AB - OBJECTIVE: To investigate the influence of dermal template on the expressions of signal transduction protein Smad 3 and transforming growth factor beta1 and its receptor during wound healing process in patients with deep burns. METHODS: Twenty burn patients with excision of full thickness burn in the extremities were enrolled in the study and divided into two groups, i.e. template interfering group (E, n = 20, grafting of dermal template [allogeneic acellular dermal matrix] with razor thin autoskin) and control group (C, n = 20, grafting of razor thin autoskin only). The contralateral side served as the self-control. Tissue samples from the burn wounds were harvested at 1, 2, 3 and 4 post-operative weeks (POW) for immunohistochemistry staining. The positive expression rates of TGF beta1, TbetaRI, TbetaRII and Smad3 proteins were determined by image analysis system. RESULTS: The positive expressions of TGFbeta1, TbetaRI, TbetaRII and signal transduction protein Smad 3 in the tissue samples in both groups could be identified during 1 approximately 4 POW, and they diminished thereafter with the process of wound healing. The expression rate of TGF-beta1 in E group was (13.08 +/- 4.65)% at 1 POW and (9.03 +/- 1.89)% at 4 POW. The positive expression rate of above indices in E group was obviously lower than that in C group in corresponding time points (P < 0.05). CONCLUSION: The expression levels of TGFbeta1, TbetaRI, TbetaRII and Smad 3 protein in deep burn wounds could be lowered by mixed grafting of dermal template with razor thin autoskin, which might be beneficial in ameliorating of scar hyperplasia in the burn wound. PMID- 15796854 TI - [Changes in the expression of apoptotic genes in the intestinal tissue of scalded rats before and after resuscitation]. AB - OBJECTIVE: To explore the dynamic changes in the expression of c-fos, proliferation cell nuclear antigen (PCNA) and Bax in the intestinal tissue of scalded rats before and after resuscitation. METHODS: Wistar rats inflicted with 30% TBSA full thickness scald were employed as the model and randomly divided into four groups with 8 in each group, i.e. 2.0, 2.5, 4.0, 6.0 postscald hour (PSH) groups. Rats in each group received routine fluid infusion at 2 PSH, and were sacrificed at 2, 2.5, 4, 6 PSH, respectively. Then the intestinal tissue of the rats was harvested for the detection of the expression of c-fos, PCNA and Bax. RESULTS: The expression of c-fos, PCNA and Bax at 2.0 PSH group (65.8 +/- 4.2%, 74.5 +/- 2.4%, 26.3 +/- 5.7%, respectively) significantly increased when compared with those in 2.5 PSH group (92.4 +/- 5.7%, 85.6 +/- 4.5%, 67.1 +/- 6.6%, respectively) (P < 0.01). The expression of 3 genes increased dramatically at 2.5 and 4.0 PSH, and reached the peak at 2.5 PSH. There was no obvious difference in the gene expression between 4 PSH and 2 PSH groups. CONCLUSION: The expression of apoptotic genes in the intestinal tissue of scalded rats increased significantly during early resuscitation stage after burn injury. PMID- 15796855 TI - [Experimental study on early escharectomy after major burns in minipigs]. AB - OBJECTIVE: To investigate optimal time for early escharectomy after severe scald in minipigs. METHODS: Minipigs inflicted with 35% TBSA full thickness burn were employed in the study, and they were randomly divided into A (n = 7, with escharectomy at 6PSH), and B (n = 7, with escharectomy at 24 PSH) groups. The hemodynamics indices, hemorrheology, and the serum levels of cytokines in the two groups were determined before burns and at 6, 8, 16, 24 and 72PSH. RESULTS: The hemodynamics indices in A group obviously improved compared with those in B group. The cardiac output (CO, 2.28 +/- 0.03 L/min) and right arterial pressure (RAP, 4.54 +/- 0.04 mmHg) in A group recovered to the pre-injury levels at 48 PSH. There was no difference of the hemorrheology indices between the two groups (P > 0.05). The serum contents of cytokines in A group declined to the pre-injury level on 1 PBW, while those in B group were significantly higher than those before injury and those in A group (P < 0.05 or P > 0.01). CONCLUSION: Escharectomy during 6 PBH might be safe and feasible, thus preventing long-term complications effectively. PMID- 15796857 TI - [Influence of hyperoxic fluid on the down-regulated proteins of intestinal mucosa in scalded rats]. AB - OBJECTIVE: To investigate the influence of hyperoxic fluid on the down-regulated proteins of intestinal mucosa in scalded rats, so as to provide a theoretical basis for the clinical use of hyperoxic fluid. METHODS: Sprague-Dawley (SD) rats with 35% TBSA full-thickness scald were randomly divided into scald control (S, n = 6,with intraperitoneal fluid infusion after scalding), hyperoxic (H, n = 6, with hyperoxic fluid infusion after scalding) groups. Six rats without scald injury served as normal group. The proteins in the intestinal mucosa were separated with the two-dimensional electrophoresis (2-DE), and were analyzed with ImageMaster 2D Elite. The influence of hyperoxic fluid on the down-regulated proteins of intestinal mucosa in scalded rats was studied with bio-spectrum, protein bank and document analysis. RESULTS: (1) Among the 34 down-regulated protein spots in S group, 9 of them definitely exhibited up-regulation compared with those in H group. (2) The expression of mitochondrial aconitase, alpha propionyl-CoA carboxylase, short chain of hydroxyacyl-Coenzyme A dehydrogenase, transcription factor EB (estradiol benzoate), triosephosphate isomerase 1, T cell receptor V delta 6, and dynein-like protein-5 in H group were significantly up regulated. CONCLUSION: The hyperoxic fluid could up-regulate the down-regulated proteins in rat intestinal mucosa at early postburn stage, so that the barrier function of intestinal mucosa of rats with severe burns could be partially recovered. PMID- 15796872 TI - [Gene detection: an impact on the development of pathology]. PMID- 15796873 TI - [Colorectal polyps and colorectal cancer]. PMID- 15796874 TI - [Gastrointestinal stromal tumor: a clinicopathological study of 74 cases]. AB - OBJECTIVE: To explore a simplified and reproducible approach for the diagnosis and morphologic prognostication of gastrointestinal stromal tumor (GIST). METHODS: Eighty-five cases of gastrointestinal mesenchymal tumors including 74 cases of GIST, 8 esophageal smooth muscle tumor, 1 rectal leiomyosarcoma, 1 Schwannoma, and 1 malignant fibrous histiocytoma were studied by histological evaluation along with an immunohistochemistry panel including vimentin, CD117 (c kit), CD34, SMA, desmin and S-100. Clinicopathological correlation was performed in 31 cases of GIST that had accompanied with the available follow-up data. RESULTS: Among 74 GISTs, 34 arose principally from the stomach, 30 from the small intestine, and 10 other cases found in the esophagus, retroperitoneum, mesenterium and omentum. The patients' age ranged from 23 to 80 years (mean 52.5 years), with 45 males and 29 females. Histologically, the tumors composed of either spindle or oval to round cells arranged in interlacing fascicles forming whorls or cellular clusters, cytoplasm generally abundant and eosinophilic. There were 48 cases of spindle cell type, 10 cases of epithelioid cell type and 16 cases of mixed cell type. All 74 cases of GIST were positive for CD117 in a cell membranous pattern, however, some variable staining patterns of CD117 had been noticed in a few cases. In addition, 54 GISTs were also positive for CD34 (72.9%), 25 cases positive for SMA, 5 cases positive for S-100 and 5 cases positive for desmin. According to the Fletcher's scheme, GISTs in this study were divided into 4 subcategories including groups of very low risk of aggressive behavior (3 cases), of low risk (15 cases), of intermediate risk (36 cases) and of high risk (20 cases) respectively. Kaplan-Meier survival analysis of 31 GIST cases whom had been followed up for 16 to 72 months showed a statistically significant difference present among the subcategories (P < 0.01). CONCLUSIONS: GISTs predominantly occur in the middle and old age patients, more common in male, and positive CD117 staining is considered to be the defining marker to differentiate GIST from other mesenchymal tumors of the GI tract. Positive CD34 immun-staining, plus a CD117 positivity, strengthens further a diagnosis of GIST. Subclassification of GISTs using Fletcher's scheme appears to be simple, reproducible, and correlates well with the clinical behavior of the tumor. PMID- 15796875 TI - [Clinicopathological and immunohistochemical study of extra-gastrointestinal stromal tumors arising from the omentum and mesentery]. AB - OBJECTIVE: To explore the clinicopathological and immunohistochemical features of extra-gastrointestinal stromal tumors (EGIST) arising from the omentum and mesentery and to investigate the cellular origin of these tumors, prognostic factors, and the relationships with gastrointestinal stromal tumors. METHODS: Nineteen cases of mesenchymal neoplasms arising from the omentum and mesentery (previously diagnosed as smooth-muscle tumors or schwannomas) were studied morphological with a panel of immunohistochemistry including CD117 and CD34. RESULTS: Among the 19 cases, 14 tumors were confirmed to be EGIST, of which 6 tumors arose from the omentum and 8 cases located at the mesentery. The size of tumors ranged from 3.5cm to 29.0 cm (mean 12.4cm) in diameter. Histologically, there were 9 cases of mainly spindle cell type, 2 cases of mainly epithelioid cell type and 3 cases of mixed cell type. all EGIST expressed CD117 (14/14) and a percentage of them expressed also CD34 (8/14) and/or SMA (6/14), anyhow, all EGIST were negative for desmin and S-100 protein. Six patients with tumors arising from the omentum were all alive without evidence of disease (tumor-free). Among 7 cases with tumors of the mesentery, three patients died of the disease, 1 alive with the disease and 3 patients alive without evidence of the disease. CONCLUSIONS: EGIST were identical by their histological and immunohistochemical features with gastrointestinal stromal tumors (GIST). This tumor may arise from the multipotential mesenchymal stem cells. EGIST have various clinical behavior, and the parameters used for predicting the prognosis of GIST may not be completely suitable for EGIST evaluation. PMID- 15796876 TI - [Monoclonal antibodies against human tumor metastasis suppressor gene-1 TMSG-1: preparation, characterization and application]. AB - OBJECTIVE: In order to clarify the exact molecular weight of tumor metastasis suppressor gene-1 (TMSG-1) protein and its cellular localization, a monoclonal antibody against TMSG-1 was prepared, characterized and applied to evaluate the metastatic potential of human tumors. METHODS: A dominant epitope-TMSG-1(15) derived from TMSG-1 was synthesized based on Fmoc method, and the hapten was conjugated to Imject Maleimide activated mcKLH as a carrier protein. The antigen preparation was used to immunize BAL B/C mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, which were further characterized by western blotting and immunohistochemical staining. RESULTS: One hybridoma cell line secreting anti-TMSG-1 antibody, designated as C8, was eventually established after primary ELISA screening, followed by rapid limited dilution procedure. It was confirmed that C8 was of IgM isotype. Result of competitive inhibition assay showed that the antibody was TMSG-1 specific. Using this antibody, an expected protein band of about 45,000 (relative molecular mass) was detected in the non-metastatic variants PC(3)-2B4 and PG-LH7 cells by Western blotting, but not in the isogenetic metastatic variants of PC3-1E8 and PG-BE1 cells. Immunohistochemistry using C8 showed a positive staining of cell membrane and cytoplasm of 2B4 and LH7 cells, whereas 1E8 and BE1 cells were non-reactive. Immunostaining using C8 of paraffin sections of 52 breast carcinomas and 41 colon cancers demonstrated a strong positivity in non-metastatic tumors, but none to weakly reactive in metastatic tumors (P < 0.05). CONCLUSION: C8 monoclonal antibody against the synthetic peptide is TMSG-1 specific and is effective for Western blot and immunohistochemistry assays to detect TMSG-1 expression in cancer cells. TMSG-1 protein is about 45 000 (relative molecular mass) at cell membrane and cytoplasm of tumor cells. Expression of TMSG-1 protein correlates well, inversely with the tumor metastatic potential. PMID- 15796877 TI - [Clinical and pathological reassessment of 493 cases of non-Hodgkin's lymphomas according to current WHO classification of lymphoid neoplasms]. AB - OBJECTIVE: To investigate the clinical and pathological features of non-Hodgkin's lymphoma (NHL) and to evaluate the applicability of the new WHO classification of lymphoid neoplasms. METHODS: According to the new WHO classification, a total of 500 cases of non-Hodgkin's lymphoma diagnosed during the period 1992 - 2003 were reviewed and reappraised with their morphological, immunological and clinical characteristics. Clinical survival analysis was performed in 156 cases that accompanied with follow-up data. RESULTS: Among 500 cases previously diagnosed as lymphomas, 493 cases (98.6%) were confirmed to be NHL, of which B-cell neoplasms was 69.0% and T/NK-cell neoplasms 29.8%. Overall, 6 subtypes including diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), unspecified peripheral T cell lymphoma (PT-un), precursor T-lymphoblastic lymphoma (T-LBL), extranodal marginal zone B-cell lymphoma of MALT type (MALT) and B-small lymphocytic lymphoma (B-SLL) were among the most common subtypes. In pediatric and young patient populations, the most common subtypes were LBL, DLBCL and Burkitt's lymphoma. The frequency of LBL in all patients, especially in the juniors, was much higher than those reported outside Mainland China, and the frequency of FL was much higher than the reported in Mainland China. The frequency of FL was much higher than the reported in Mainland China. Clinical survivals among different histological subtypes of NHL varied considerably with statistic significance (P < 0.001). Marginal zone B-cell lymphoma and SLL demonstrated the best prognosis, LBL and PT-un both the worst, whereas DLBCL and FL had an intermediate prognosis, however, subgrouping of FL according to WHO classification did not reveal a significant survival difference (P > 0.05). CONCLUSIONS: Basing upon the results of a comprehensive survey on the morphologic features, immunophenotyping and clinical data of the above cases, the new WHO classification of lymphoid neoplasms is practical and easily applicable for routine pathological evaluation of lymphoproliferaive disorders and in guiding the clinical management. It appears that the diagnostic and grading criteria for FL in Mainland China need to be re-evaluated. PMID- 15796878 TI - [Detection of FUS-CHOP fusion gene in paraffin-embedded tissues and its clinicopathologic significance for myxoid/round cell liposarcomas]. AB - OBJECTIVE: To explore the feasibility of detecting FUS-CHOP fusion gene in formalin-fixed, paraffin-embedded tissue and its application in the diagnosis and differential diagnosis of myxoid/round cell liposarcomas (MRCLs). METHODS: Forty four formalin-fixed, paraffin-embedded MRCL samples and 60 control cases (atypical/well-differentiated liposarcoma, pleomorphic liposarcoma, low-grade myofibrosarcoma, etc.) retrieved from the archival files were studied. Nested reverse transcription-polymerase chain reaction (RT-PCR) technique was employed to detect the FUS-CHOP mRNA expression, followed by DNA sequencing confirmation of the PCR product. Housekeeping gene PGK was used to assess the quality of the mRNA templates. RESULTS: PGK mRNA was detected in 93 of 104 tumor cases (89.4%), including 39 MRCLs cases (39/44, 88.6%) and 90% of the negative control cases. Type II FUS-CHOP fusion transcript was successfully detected in 20 out of 39 (51.3%) MRCL cases. Type I FUS-CHOP fusion transcript was not detected in any MRCLs in this study. All 60 negative control cases were negative for the FUS-CHOP fusion gene transcripts. CONCLUSIONS: (1) Nested RT-PCR can be used to detect FUS CHOP mRNA in formalin-fixed, paraffin-embedded tissues. (2) FUS-CHOP is considered a specific molecular and genetic hallmark for MRCLs. Nested RT-PCR is a sensitive and specific technique in detecting FUS-CHOP gene, and can be used in the diagnosis and differential diagnosis of MRCLs. PMID- 15796879 TI - [Clinicopathologic and immunohistochemical study of pulmonary epithelioid hemangioendothelioma]. AB - OBJECTIVE: To study the clinical and pathological characteristics of pulmonary epithelioid hemangioendothelioma. METHODS: Four cases of pulmonary epithelioid hemangioendothelioma were studied by histopathologic and immunohistochemical examination of lung biopsy specimens. RESULTS: There were 3 female and 1 male, age 28 to 40 years. Clinically the tumor presented as multiple bilateral small nodules in the lung. Histologically, crown-like clusters of epithelioid tumor cells were obtained which filled in the alveoli locating at the periphery of the tumor nodules, while the central part of the nodules contained myxoid to hyaline matrix. The overall architecture of the lung was still preserved. Additionally, intracytoplasmic vacuoles were seen in tumor cells within which red blood cells were sometimes identified. Tumor cells generally lacked pleomorphism, mitotic activity and necrosis. They were immunohistochemically positive for CD31 and CD34. AE1/AE3 staining was positive in some cases. CONCLUSIONS: Pulmonary epithelioid hemangioendothelioma often occurs in a middle-aged woman and represents a distinct clinical pathological entity. PMID- 15796880 TI - [Clinical significance of heparanase and basic fibroblast growth factor expression in human non-small cell lung cancer]. AB - OBJECTIVE: To assess protein and mRNA expression levels of heparanase and basic fibroblast growth factor (bFGF) genes in human non-small cell lung cancer (NSCLC) and their roles in tumor invasion, metastasis and prognosis. METHODS: A total of 115 paraffin-embedded and 45 fresh-frozen tissue specimens of NSCLC were studied by immunohistochemistry, Western Blot and in situ hybridization to evaluate the protein and mRNA expression status of heparanase and bFGF genes. The data was analyzed by SPSS statistical software. RESULTS: Both human heparanase and bFGF were highly expressed in NSCLC cells, in contrast to none or a low expression in normal lung tissue. Expression of heparanase also showed a significantly higher than that in the normal tissue by Western blot (P = 0.041). Immunohistochemistry showed that heparanase expression was both cytoplasmic and membranous. The agreement between heparanase and bFGF was significant. A significant correlation was found between the expression of either protein and TNM stage, vascular invasion, lymphatic metastasis and microvascular density (MVD). Co-expression of the two proteins demonstrated an even higher correlation with the tumor stage and MVD. In addition, expression of bFGF correlated with tumor cell differentiation. Data of a multivariate analysis indicated that tumor cell differentiation, vascular invasion, lymphatic metastasis and expression of bFGF were identified as significant prognostic parameters. CONCLUSIONS: Both heparanase and bFGF may play important roles in tumor angiogenesis, metastasis, and prognosis of NSCLC. PMID- 15796882 TI - [Quantum dots: a novel type of fluorescent labelling substances]. PMID- 15796881 TI - [Malignant transformation of NIH3T3 cells induced by ectopic expression of PC-1 gene]. AB - OBJECTIVE: To establish a mouse fibroblastic cell line stably transfected with PC 1 gene, and using such cell line to investigate tumor development and progression imposed by the ectopic expression of PC-1 gene. METHODS: Eukaryotic expression vector pcDNA3.1(-)/myc-his-pc-1 was transfected into mouse fibroblast cell line NIH3T3 by lipofectamine. Stable transfectants were selected by G418. The integration and expression of ectopic PC-1 gene were analyzed by PCR and RT-PCR. Cytomorphological analysis, MTT, soft agar colony formation and nude mice tumorigenesis assay were used to evaluate the effects of PC-1 gene expression on tumor development and progression. RESULTS: NIH 3T3 cell lines stably expressing PC-1 gene were successfully established and confirmed by PCR and RT-PCR analyses of the integration and expression of ectopic PC-1 gene. Comparing with the parental cell line and cells transfected with control vector, the PC-1 gene transfectants acquired several phenotypes of transformed cells: increasing growth rate, ability to grow and form cell colonies on soft agar, and becoming tumorigenic in nude mice. CONCLUSION: Ectopic expression of PC-1 gene in NIH3T3 cells can induce malignant transformation of mouse fibroblastic cells both in vitro and in vivo. PMID- 15796883 TI - [Standardization of CD117 expression in the diagnosis of gastrointestinal stromal tumor]. PMID- 15796884 TI - [Influence of different antigen retrieval on the immunohistochemistry]. PMID- 15796886 TI - [Congenital malignant melanoma in a neonate]. PMID- 15796885 TI - [Spindle cell variants of diffuse large B cell lymphoma: report of 2 cases]. PMID- 15796887 TI - [A massive chest wall myxoid chondrosarcoma protruding into the thoracic cavity]. PMID- 15796888 TI - [Cytophagic histiocytic panniculitis]. PMID- 15796890 TI - The correlation between genomic G+C and optimal growth temperature of prokaryotes is robust: a reply to Marashi and Ghalanbor. AB - We have recently shown that optimal growth temperature (T(opt)) is one of the factors that influence genomic GC in prokaryotes. Our results have been disputed by Marashi and Ghalanbor, who claim that the correlations we show are not "robust" because the elimination of some points (arbitrarily chosen) leads, in some families, to variations in the correlation coefficients and/or significance of correlations. Here, we test whether the correlation between T(opt) and genomic GC is robust by using two independent approaches: detection of possible outliers (using robust Mahalanobis distance) and usage of a non-parametric correlation coefficient that is not sensitive to the presence of outliers. The results presented here reinforce our previous proposal that T(opt) is correlated with genomic GC in prokaryotes. PMID- 15796891 TI - Glyceraldehyde-derived advanced glycation end-products preferentially induce VEGF expression and reduce GDNF expression in human astrocytes. AB - The blood-brain barrier (BBB) is a biological unit composed of capillary endothelial cells and astrocytes. Here we examined the effects of various types of advanced glycation end-products (AGEs) on astrocytes and BBB-forming endothelial cells. While no type of AGE we examined changed the permeability of endothelial sheets, glyceraldehyde-derived AGE induced VEGF expression most significantly in astrocytes. The expression of glial cell line-derived neurotrophic factor (GDNF), which reduces the vascular permeability, was decreased in the astrocytes by treatment with glyceraldehyde-derived AGE. These results indicate that glyceraldehyde-derived AGE is the biologically active substance for astrocytes by regulating the VEGF and GDNF expression, which is causally contributing to an increase in the permeability of the BBB. PMID- 15796892 TI - 19F NMR measurements of NO production in hypertensive ISIAH and OXYS rats. AB - Recently we demonstrated the principal possibility of application of 19F NMR spin trapping technique for in vivo *NO detection [Free Radic. Biol. Med. 36 (2004) 248]. In the present study, we employed this method to elucidate the significance of *NO availability in animal models of hypertension. In vivo *NO-induced conversion of the hydroxylamine of the fluorinated nitronyl nitroxide (HNN) to the hydroxylamine of the iminonitroxide (HIN) in hypertensive ISIAH and OXYS rat strains and normotensive Wistar rat strain was measured. Significantly lower HIN/HNN ratios were measured in the blood of the hypertensive rats. The NMR data were found to positively correlate with the levels of nitrite/nitrate evaluated by Griess method and negatively correlate with the blood pressure. In comparison with other traditionally used methods 19F NMR spectroscopy allows in vivo evaluation of *NO production and provides the basis for in vivo *NO imaging. PMID- 15796893 TI - Involvement of lipid peroxidation in the degradation of a non-phenolic lignin model compound by manganese peroxidase of the litter-decomposing fungus Stropharia coronilla. AB - Culture liquids of the litter-decomposing basidiomycete Stropharia coronilla showed pro-oxidant activity promoting the peroxidation of linoleic acid. This activity depended on the presence of manganese peroxidase (MnP) in the fungal culture. Pro-oxidant activity maxima coincided with maximum MnP activities during the separation of extracellular proteins by anion-exchange chromatography. Purified MnP1 showed substantial pro-oxidant activity in the presence of acetate and Mn2+ ions, even without the addition of hydrogen peroxide. A non-phenolic beta-O-4 lignin model compound [LMC; 1-(3,4-dimethoxyphenyl)-2-(2-methoxyphenoxy) 1,3-dihydroxypropane] was partially oxidized in an in vitro reaction system developing MnP-dependent lipid peroxidation. The chelating organic acids malonate and tartrate noticeably inhibited both the peroxidation of linoleic acid and the conversion of LMC in the system. The major product of the LMC oxidation was 1 (3,4-dimethoxyphenyl)-1-oxo-2-(2-methoxyphenoxy)-3-hydroxypropane; in addition, small amounts of 3,4-dimethoxybenzaldehyde (veratraldehyde) and 3,4 dimethoxybenzoic (veratric) acid were detected. Thus, MnP-initiated lipid peroxidation may be involved in the degradation of recalcitrant non-phenolic lignin substructures by litter-decomposing fungi similar to MnPs of wood-decaying fungi. PMID- 15796894 TI - Overexpression of Bop3 confers resistance to methylmercury in Saccharomyces cerevisiae through interaction with other proteins such as Fkh1, Rts1, and Msn2. AB - We found that overexpression of Bop3, a protein of unknown function, confers resistance to methylmercury in Saccharomyces cerevisiae. Bmh2, Fkh1, and Rts1 are proteins that have been previously shown to bind Bop3 by the two-hybrid method. Overexpression of Bmh2 and the homologous protein Bmh1 confers resistance to methylmercury in yeast, but overexpression of either Fkh1 or Rts1 has a minimal effect. However, the increased level of resistance to methylmercury produced by overexpression of Bop3 was smaller in Fhk1-deleted yeast as compared with that of the wild-type strain. In contrast, the degree of resistance was significantly elevated in Rts1-deleted yeast. Msn2 and Msn4 were previously reported as proteins that bind to Bmh1 and Bmh2. Overexpression of Msn2 conferred a much greater sensitivity to methylmercury in yeast, while deletion of the corresponding gene lowered the degree of resistance to methylmercury induced by overexpression of Bop3. These results suggest that multiple proteins are involved in minimizing the toxicity of methylmercury induced by overexpression of Bop3. PMID- 15796896 TI - Feedback regulation of bile acid synthesis in human liver: importance of HNF 4alpha for regulation of CYP7A1. AB - A great number of nuclear factors are involved in the negative feedback mechanism regulating bile acid synthesis. There are two major ways for the negative feedback to effect the synthesis; the SHP-dependent, involving FXR, and the SHP independent way, affecting HNF-4alpha. We studied 23 patients with gallstone disease. Eight patients were treated with chenodeoxycholic acid, 7 with cholestyramine prior to operation, and 8 served as controls. Liver biopsies were analyzed with Real-time-PCR. In the cholestyramine-treated group mRNA levels of CYP7A1 were increased about 10-fold. Treatment with CDCA decreased the mRNA levels of CYP7A1 by about 70%. The mRNA levels of CYP8B1, CYP27A1, and CYP7B1 were not significantly altered in the treated groups. The analysis of mRNA levels for HNF-4alpha showed 64% higher levels in the cholestyramine-treated group compared to the controls. These levels showed positive and highly significant correlation to the levels of mRNA of CYP7A1 when studied in all three groups together. FXR, SHP, and LRH-1/FTF were not significantly affected by the different treatments. Our results indicate that when bile acid synthesis is upregulated by cholestyramine treatment the SHP-independent pathway for controlling CYP7A1 transcription dominates over the SHP-dependent pathway. PMID- 15796895 TI - Systemic hypoxia alters gene expression levels of structural proteins and growth factors in knee joint cartilage. AB - We investigated the effects of short- (8- and 24-h) and long-term (3 weeks) exposure to systemic normobaric hypoxia (13%) on the gene expression level of structural proteins and growth factors in knee joint cartilage of rabbits. Collagen type Ia2, II, and Va1, TGF-beta1, and b-FGF were upregulated after short term hypoxia in both menisci, but not in articular cartilage. In contrast, long term hypoxia downregulated gene expression level of collagens, aggrecan, and growth factors in articular cartilage and meniscal fibrocartilage. Interestingly, gene expression levels of non-collagenous proteins biglycan, decorin, and versican were not affected by short-term or by long-term hypoxia in knee joint cartilage. The present study suggests that changes in oxygen level differentially affect gene expression levels of growth factors, collagens, and non-collagenous proteins in normal knee joint cartilage in rabbits. PMID- 15796897 TI - Alkaline phosphatase retained in HepG2 hepatocarcinoma cells vs. alkaline phosphatase released to culture medium: difference of aberrant glycosylation. AB - Liver tissue is the source of 90% of serum alkaline phosphatase (AP). The serum levels and structures of tumor marker proteins change under many disease conditions as well as cancer. The study was aimed at determining the type of alkaline phosphatase (AP) present in HepG2 hepatocellular carcinoma cell line. Alkaline phosphatase rich extracts of healthy human liver, HepG2 hepatocarcinoma cells, as well as the condition medium of HepG2 cells were prepared by extraction with 40% n-butanol and 30-50% acetone precipitation, and subjected to various chromatographic procedures. Lectin affinity chromatography of the samples with concanavalin A-Sepharose 4B showed considerable differences in the elution patterns. Non-denaturing polyacrylamide gel electrophoresis of the culture medium yielded a relatively slow migrating band of activity that coincided with none of the three bands of activity produced by the normal liver extract, nor with the bands of the cell pellet extract. Inhibition patterns were established by measuring the enzyme activities in the presence of varying concentrations of L phenylalanine, L-leucine, L-homoarginine, and levamisole. The APs from the cell line were neuraminidase sensitive. According to the results the main AP produced and released to the medium by HepG2 cell line is an aberrantly glycosylated tissue non-specific AP. In addition, the differences between the cell-pellet AP and the culture medium AP seemed to stem from different sugar moieties in their structures. PMID- 15796898 TI - Regulation of apical localization of the thiazide-sensitive NaCl cotransporter by WNK4 in polarized epithelial cells. AB - Missense mutations in the WNK4 gene have been postulated to cause pseudohypoaldosteronism type II (PHAII), an autosomal-dominant disorder characterized by hyperkalemia and hypertension. Previous reports using Xenopus oocytes showed that wild-type WNK4 expression inhibited surface expression of the thiazide-sensitive NaCl cotransporter (NCC), while a disease-causing mutant lost the inhibitory effect on NCC surface expression. To determine if these changes observed in oocytes really occur in polarized epithelial cells, we generated stable MDCK II cell lines expressing NCC alone or NCC plus wild-type WNK4 or a disease-causing (D564A) WNK4. In contrast to the apical localization of NCC without co-expression of WNK4, immunofluorescence microscopy and biotin surface labeling revealed that this apical localization was equally decreased by both the wild-type and the mutant WNK4 expression. Apical localizations of two PHAII unrelated apical transporters, sodium-independent amino acid transporter, BAT1 and bile salt export pump, Bsep, were also found to be decreased by both wild type and mutant WNK4 expression. These results indicate that the regulation of NCC was not related to the disease-causing mutation and not restricted to the PHAII-related specific transporters. The regulation of intracellular localization of NCC by WNK4 might not be involved in the pathogenesis of PHAII. PMID- 15796899 TI - In vivo imaging of oxidative stress in the kidney of diabetic mice and its normalization by angiotensin II type 1 receptor blocker. AB - This study was undertaken to evaluate oxidative stress in the kidney of diabetic mice by electron spin resonance (ESR) imaging technique. Oxidative stress in the kidney was evaluated as organ-specific reducing activity with the signal decay rates of carbamoyl-PROXYL probe using ESR imaging. The signal decay rates were significantly faster in corresponding image pixels of the kidneys of streptozotocin-induced diabetic mice than in those of controls. This technique further demonstrated that administration of angiotensin II type 1 receptor blocker (ARB), olmesartan (5 mg/kg), completely restored the signal decay rates in the diabetic kidneys to control values. In conclusion, this study provided for the first time the in vivo evidence for increased oxidative stress in the kidneys of diabetic mice and its normalization by ARB as evaluated by ESR imaging. This technique would be useful as a means of further elucidating the role of oxidative stress in diabetic nephropathy. PMID- 15796900 TI - Identification of parathyroid hormone-regulated proteins in mouse bone marrow cells by proteomics. AB - The ability of parathyroid hormone (PTH) to enhance bone formation has recently been exploited in the treatment of osteoporosis. Several studies have suggested that the activation of bone marrow stromal cells could be preceded to show the anabolic effect of PTH on bone formation, but little is known of PTH-regulated proteins in bone marrow cells. Therefore, protein profiling in the intermittent PTH-treated bone marrow cells was evaluated using proteomics. Daily treatment for 5 days consisting of subcutaneous injection of either 150 microg/kg per day of mouse PTH (1-84) or vehicle (0.9% normal saline) was performed on the ICR mouse. At the end of the treatment period, bone marrow cells were separated and used in proteomics. The expression levels of seven proteins including vimentin were decreased, but those of four proteins including calreticulin and thioredoxin domain containing 7 protein (Txnde7) were increased. Among these, the decrease of vimentin and the increase of both calreticulin Txnde7 in mRNA levels were confirmed by semi-quantitative RT-PCR. In PTH-treated mouse MC3T3-E1 osteoblast cells, mRNA expression levels were not totally consistent with the results observed in proteomics. In conclusion, the differentially expressed proteins in bone marrow cells depending on PTH could be highly linked to the differentiation of osteoprogenitor cells in the bone marrow into preosteoblast cells. PMID- 15796901 TI - Sphingolipids are involved in N-methyl-N'-nitro-N-nitrosoguanidine-induced epidermal growth factor receptor clustering. AB - Previously we have found that N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), an alkylating agent, can induce the clustering of cellular surface receptors including tumor necrosis factor receptor (TNFR) and epidermal growth factor receptor (EGFR). Since sphingolipids, especially ceramide, have been suggested as major players in ligand-induced receptor clustering, their involvement in this ligand-independent, chemical-induced receptor clustering was evaluated. It was shown that MNNG-induced EGFR clustering occurred primarily at lipid rafts, as nystatin, which can disrupt lipid raft structure, significantly decreasing MNNG induced EGFR clustering. Lipidomic studies revealed that MNNG treatment induced profound changes in sphingolipids metabolism, which were not the same as those induced by EGF treatment. Acid sphingomyelinase (ASM) is responsible for hydrolyzing sphingomyelin to generate ceramide, and it was demonstrated that MNNG treatment caused ASM distribution changing from diffused state to concentrated area of cells, which colocalized with lipid rafts. Nystatin treatment also abolished the redistribution of ASM. In addition, blockage of ceramide production by ASM inhibitor imipramine interrupted MNNG-induced receptor clustering. Taken together, these data suggested that sphingolipids are involved in MNNG-induced receptor clustering; however, the specific species involved may be different from those involved in EGF-mediated receptor clustering. PMID- 15796903 TI - Receptor related to tyrosine kinase RYK regulates cell migration during cortical development. AB - Mammalian RYK is a receptor related to tyrosine kinase without detectable catalytic activity. We have previously reported that rat RYK is dominantly expressed in neural progenitor cells and mature neurons in the developing central nervous system. Mouse RYK has been found to bind to EphB2/B3 receptors, which have diverse functions during development. In this study, we demonstrated that RYK, EphB2, EphB3, ephrinB1, and ephrinB2 are expressed in embryonic brain. In vitro analysis using COS-7 cells revealed binding between rat RYK and EphB3, and that the RYK deletion mutant without extracellular leucine-rich motifs lacked this binding ability. To investigate the function of RYK in vivo, embryonic cortical slice cultures were analyzed after electroporation of expression plasmids for RYK or its deletion mutants. The results showed that overexpression of RYK suppressed cell migration from the ventricular zone to the pial surface, however, overexpression of the RYK deletion mutant without leucine-rich motifs had no effect on cell migration. These results suggest that RYK regulates cell migration during mammalian cortical development through the binding to Eph receptors. PMID- 15796902 TI - The heterodimer of alpha4 and PP2Ac is associated with S6 kinase1 in B cells. AB - Alpha4 is a signal transduction molecule that is required for B cell activation. Alpha4 associates with the catalytic subunit of protein phosphatase 2A (PP2Ac) and regulates its enzymatic activity. We examined the interaction of alpha4/PP2Ac with S6 kinase1 (S6K1) as a potential downstream signal transduction molecule because both alpha4/PP2Ac association and S6K1 activity were rapamycin-sensitive. Stimulation of spleen B cells with lipopolysaccharide induced the interaction of alpha4/PP2Ac and S6K1. Pull-down assay demonstrated that alpha4 interacts with S6K1 through PP2Ac. S6K1 and alpha4 bind to the different regions of PP2Ac as S6K1 to the region from amino acid 88th to 309th of PP2Ac and alpha4 to the two separated regions of the amino-terminal (from amino acid 19th to 22nd) and the middle (from 150th to 164th) portions of PP2Ac. These results suggest that alpha4 regulates S6K1 activity through PP2Ac in B cell activation. PMID- 15796904 TI - Caspase-3 sensitive signaling in vivo in apoptotic HeLa cells by chemically engineered intramolecular fluorescence resonance energy transfer mutants of green fluorescent protein. AB - Green fluorescent protein (UV5) was re-engineered to remove native cysteine residues, and a new cysteine was introduced near the C-terminus, approximately 20 A from the native fluorophore, for site-specific attachment of chemical fluorophores. The resultant efficient intramolecular FRET quenched GFP emission and gave a new emission band from the conjugated fluorophore. Caspase-3 cleavage of constructs with a caspase-3 sequence near the C-terminus in the sequence between the native fluorophore and the new cysteine, located C-terminal to the caspase site, destroyed the FRET, the emitted color reverting to that of unmodified GFP. This process was demonstrated in vitro with caspase-3 and lysates from cells undergoing apoptosis. Real-time emission changes for the Alexa Fluor 532 conjugate of this GFP, studied quantitatively in vivo for single HeLa cells using the ratios of fluorescence at the red and green maxima by confocal microscopy, showed that caspase-3 action in the cytosol preceded that in the nucleus. PMID- 15796905 TI - Hepatic steatosis in transgenic mice overexpressing human histone deacetylase 1. AB - It is generally thought that histone deacetylases (HDACs) play important roles in the transcriptional regulation of genes. However, little information is available concerning the specific functions of individual HDACs in disease states. In this study, two transgenic mice lines were established which harbored the human HDAC1 gene. Overexpressed HDAC1 was detected in the nuclei of transgenic liver cells, and HDAC1 enzymatic activity was significantly higher in the transgenic mice than in control littermates. The HDAC1 transgenic mice exhibited a high incidence of hepatic steatosis and nuclear pleomorphism. Molecular studies showed that HDAC1 may contribute to nuclear pleomorphism through the p53/p21 signaling pathway. PMID- 15796906 TI - The pyrimidinergic P2Y6 receptor mediates a novel release of proinflammatory cytokines and chemokines in monocytic cells stimulated with UDP. AB - The human P2Y6 receptor (hP2Y6) is a member of the G protein-coupled pyrimidinergic P2 receptor family that responds specifically to the extracellular nucleotide uridine diphosphate (UDP). Recently, the hP2Y6 receptor has been reported to mediate monocyte IL-8 production in response to UDP or lipopolysaccharide (LPS), but the role of hP2Y6 in regulating other pro inflammatory cytokines or mediators is largely unknown. We demonstrate here that UDP specifically induces soluble TNF-alpha and IL-8 production in a promonocytic U937 cell line stably transfected with hP2Y6. However, we did not detect IL 1alpha, IL-1beta, IL-6, IL-10, IL-18, and PGE2 in the conditioned media from the same cell line. These results distinguish UDP/P2Y6 signaling from LPS signaling. Interestingly, UDP induces the production of IL-8, but not TNF-alpha, in human astrocytoma 1321N1 cell lines stably transfected with hP2Y6. Therefore, the immune effect of UDP/P2Y6 signaling on the production of proinflammatory cytokines is selective and dependent on cell types. We further identify that UDP can also induce the production of proinflammatory chemokines MCP-1 and IP-10 in hP2Y6 transfected promonocytic U937 cell lines, but not astrocytoma 1321N1 cell lines stably transfected with hP2Y6. From the Taqman analysis, UDP stimulation significantly upregulates the mRNA levels of IL-8, IP-10, and IL-1beta, but not TNF-alpha. Taken together, these new findings expand the pro-inflammatory biology of UDP mediated by the P2Y6 receptor. PMID- 15796907 TI - Dimerize RACK1 upon transformation with oncogenic ras. AB - From our previous studies, we learned that syndecan-2/p120-GAP complex provided docking site for Src to prosecute tyrosine kinase activity upon transformation with oncogenic ras. And, RACK1 protein was reactive with syndecan-2 to keep Src inactivated, but not when Ras was overexpressed. In the present study, we characterized the reaction between RACK1 protein and Ras. RACK1 was isolated from BALB/3T3 cells transfected with plasmids pcDNA3.1-[S-ras(Q61K)] of shrimp Penaeus japonicus and RACK1 was revealed to react with GTP-K(B)-Ras(Q61K), not GDP-K(B) Ras(Q61K). This selective interaction between RACK1 and GTP-K(B)-Ras(Q61K) was further confirmed with RACK1 of human placenta and mouse RACK1-encoded fusion protein. We found that RACK1 was dimerized upon reaction with GTP-K(B)-Ras(Q61K), as well as with 14-3-3beta and geranylgeranyl pyrophosphate, as revealed by phosphorylation with Src tyrosine kinase. We reported the complex of RACK1/GTP K(B)-Ras(Q61K) reacted selectively with p120-GAP. This interaction was sufficient to dissemble RACK1 into monomers, a preferred form to compete for the binding of syndecan-2. These data indicate that the reaction of GTP-K(B)-Ras(Q61K) with RACK1 in dimers may operate a mechanism to deplete RACK1 from reaction with syndecan-2 upon transformation by oncogenic ras and the RACK1/GTP-Ras complex may provide a route to react with p120-GAP and recycle monomeric RACK1 to syndecan-2. PMID- 15796908 TI - Abnormal stability of wild-type p53 protein in a human lung carcinoma cell line. AB - We report here that ectopically expressed wild-type p53 protein showed more than 6 times longer half-life than normal human fibroblasts in NCl-H1299, a widely used cell line derived from non-small cell lung carcinoma lacking the expression of p53 protein. We found no abnormality in the phosphorylation and ubiquitination of p53, and the expression levels of MDM2. Although proteasome activity measured in vitro was not significantly different between the tumor cell line and normal human fibroblasts, proteasome inhibitors, ALLN, MG115, and MG132, did not accumulate p53 protein in the tumor cell line, but did accumulate p53 in normal human cells. These results provide a novel mechanism, by which p53 is stabilized in tumor cells, and they suggest that a mediator should exist between ubiquitinated p53 and proteasome, which may be defective in H1299 cells. PMID- 15796909 TI - Integrative genomics based identification of potential human hepatocarcinogenesis associated cell cycle regulators: RHAMM as an example. AB - DNA microarray has been widely used to examine gene expression profile of different human tumors. The information generated from microarray analysis usually represents the overall range of cancer-associated abnormality associated with gene regulation. In order to identify key regulatory genes involved in carcinogenesis of human cancer, hypothesis driven data mining of the microarray data plus experimental validation becomes a critical approach in the post-genome era. Here, we present an integrative genomic analysis of published microarray data and homolog gene database. Over 20,000 genes were examined to reveal 16 genes specific to vertebrates, cell cycle G2/M regulated, and overexpressed in human HCC. Using Affymetrix microarray analysis, we found that all 16 genes were up-regulated in human HCC. Among these 16 genes, we experimentally validated the up-regulation of receptor for hyaluronan-mediated motility (RHAMM) in different cell model systems. We first confirmed elevation of RHAMM in the G2/M phase of synchronized HeLa cells. We also found that RHAMM had an elevated level of expression in all the HCC samples we examined and it was induced during the G2/M phase of regenerating mouse hepatocytes after partial hepatectomy. Thus, the expression of RHAMM appears to be tightly regulated during mammalian cell cycle G2/M progression. The ectopic overexpression of RHAMM in 293T cells resulted in the accumulation of cells at G2/M phase. RHAMM-induced mitotic arrest of cells was predominantly in the prophase. Taken together, using an integrated functional genomic approach, we have uncovered a set of genes that may play specific roles in cell cycle progression and in HCC development. To elucidate the function of these genes in cell cycle regulation may shed light on the control mechanism of human HCC in the future. PMID- 15796910 TI - Inter-individual variation of several cytochrome P450 2D6 splice variants in human liver. AB - To examine the possibility that inter-individual differences in splicing partially explain the observed differences in CYP2D6 activity, we amplified its full-length cDNA in 96 human liver RNA samples and discovered five splice variants: intron 5 retention, intron 6 retention, intron 5 and intron 6 double retention, exon 3 skipping, and partial intron 1 retention. All of the CYP2D6 splice variants we identified are probably nonfunctional transcripts. Substantial inter-individual variation in the proportions of the CYP2D6 transcript represented by splice variants, measured by real-time PCR, suggests that the presence of these splice variants contributes to the population variation in CYP2D6 activity. Relatively high levels of intron 6 retention were not correlated with the newly discovered single nucleotide polymorphism 2988G > A in intron 6 (CYP2D6*41) but did correlate with the more common CYP2D6*34 allele. Our study prompts further investigations to explore the effect of these splice variants on drug metabolism. PMID- 15796911 TI - Wnt5b partially inhibits canonical Wnt/beta-catenin signaling pathway and promotes adipogenesis in 3T3-L1 preadipocytes. AB - To elucidate the functional roles of Wnt5b in adipogenesis, we characterized gene expression profiles in Wnt5b overexpressing 3T3-L1 cells using microarray analysis. Of the approximately 20,000 genes screened, we found that 85 genes were up-regulated and 211 genes were down-regulated in 3T3-L1 cells overexpressing Wnt5b. Among the genes regulated by Wnt5b, the expressions of insulin like growth factor-1 (IGF-1), vascular endothelial growth factor-C (VEGF-C), and WNT1 inducible signaling pathway protein 1 (WISP-1), which were known to be up regulated by Wnt1/beta-catenin signaling, were decreased in the Wnt5b overexpressing cells. This result was subsequently confirmed by real-time quantitative RT-PCR (IGF-1; 0.74+/-0.08 and 0.56+/-0.08, WISP-1; 0.71+/-0.03 and 0.56+/-0.08, and VEGF-C; 0.67+/-0.01 and 0.80+/-0.07, mean+/-SEM, compared with the control at zero and two days after induction of differentiation, respectively). We also found that Wnt5b overexpression in 3T3-L1 preadipocytes was able to partially prevent the inhibitory effect of Wnt3a on adipogenesis. Furthermore, the overexpression of Wnt5b was able to inhibit Wnt3a-induced activation of the canonical Wnt/beta-catenin pathway as evidenced by the reduced translocation of beta-catenin into the nucleus. These findings indicate that Wnt5b may promote adipogenesis in 3T3-L1 cells, at least in part, by antagonizing the canonical Wnt/beta-catenin pathway. PMID- 15796912 TI - MKP-8, a novel MAPK phosphatase that inhibits p38 kinase. AB - Intracellular signaling pathways and their relationship to malignant progression have become a major focus of cancer biology. The dual-specificity phosphatase (DSP) family is a more recently identified family of intracellular signaling modulators. We have identified a novel protein phosphatase with a well-conserved DSP catalytic domain containing the DSP catalytic motif, xHCxxGxSRS, and mitogen activated protein kinase phosphatase (MKP) motif, AYLM. Because of these unique characteristics, the protein was named mitogen-activated protein kinase phosphatase-8 (MKP-8). This protein is approximately 20kDa in size and mainly localizes to the nuclear compartment of the cell. MKP-8 is expressed in embryonal cancers (retinoblastoma, neuroepithelioma, and neuroblastoma) and has limited expression in normal tissues. MKP-8 displays significant phosphatase activity that is inhibited by a cysteine to serine substitution in the catalytic domain. When co-expressed with activated MAPKs, MKP-8 is able to inhibit p38 kinase phosphorylation and downstream activity. PMID- 15796914 TI - Proteomics-based identification of DEAD-box protein 48 as a novel autoantigen, a prospective serum marker for pancreatic cancer. AB - Patients with cancer frequently develop autoantibodies, and the identification of panels of tumor autoantigens may have utility in early cancer diagnosis and immunotherapy. This study aims to exploit the autoantibody repertoire in pancreatic cancer and identify the possible serum marker for pancreatic cancer. Sera from 55 newly diagnosed patients with pancreatic cancer and 52 healthy controls were analyzed for antibody-based reactivity against Hep-2, a human larynx epithelioma cancer cell line, with one-dimensional immunoblot assay. From this analysis, we observed a prominent band with a molecular weight of 47 kDa in 63.64% (35/55) patients, while in only 1.9% normal group (1/52). Using immunoblot analysis after two-dimensional electrophoresis combined with liquid chromatography-electrospray ionization tandem mass spectrometry, this target antigen was identified as DEAD-box protein 48 (DDX48). BLAST analysis showed that it was highly similar to eukaryotic initiation factor 4A and might play a role in pre-mRNA processing. An enzyme-linked immunosorbent assay was performed using recombinant, purified DDX48 as an antigen to detect anti-DDX48 autoantibodies in sera. Reactivity was observed in 20 of 60 (33.33%) pancreatic cancer patients, 3 of 30 (10.00%) colorectal cancer patients, 2 of 30 (6.67%) gastric cancer patients, 2 of 30 (6.67%) hepatocellular cancer patients, while none of the 20 chronic pancreatitis patients, 30 lung cancer patients, and 60 normal individuals. Together, these results demonstrate that the detection of autoantibodies to DDX48 may have clinical utility for the improved diagnosis of pancreatic cancer. PMID- 15796915 TI - Excision of foreign gene product with cathepsin D in chicken hepatoma cell line. AB - To easily and rapidly recover exogenous gene products from chicken egg yolk, we constructed pVTG-catD (VTG, vitellogenin; catD, cathepsin D), a vector cassette carrying two catD-recognition signal peptides (catD-RSPs) in addition to the cloning site. An enhanced green fluorescence protein (EGFP)-encoding DNA fragment was ligated into the pVTG-catD. When the resultant construct pVTG-EGFP-catD containing histidine- and myc-tags was transfected into the chicken hepatoma cell line LMH, EGFP-expression at 24h post-cultivation was confirmed by fluorescence microscopy. Because a signal peptide (NTVLAEF) encoded in pVTG-EGFP-catD is recognized by catD, the VTG-EGFP fusion protein digested with catD was detectable by Western blotting. Digested exogenous gene product was recovered with nickel resin. These results indicate that catD-recognition sites bearing pVTG-catD and His-tags are functional in chicken LMH cells. Therefore, the system described here may be of use in making excision exogenous gene products in the chicken and in creating homozygous knock-in chickens. PMID- 15796913 TI - Structural requirements of the unique disulphide bond and the proline-rich motif within the alpha4-alpha5 loop for larvicidal activity of the Bacillus thuringiensis Cry4Aa delta-endotoxin. AB - Both the disulphide bond (Cys192-Cys199) and the proline-rich motif (Pro193ProAsnPro196) in the long loop connecting the alpha4-alpha5 transmembrane hairpin of the Cry4Aa mosquito-larvicidal protein have been found to be unique among the Bacillus thuringiensis Cry delta-endotoxins. In this study, their structural requirements for larvicidal activity of the Cry4Aa toxin were investigated. C192A and C199A mutant toxins were initially generated and over expressed in Escherichia coli cells as 130-kDa protoxins at levels comparable to that of the wild-type toxin. When their activities against Aedes aegypti larvae were determined, Escherichia coli cells expressing each mutant toxin retained the high-level toxicity. Further mutagenic analysis of the PPNP motif revealed that an almost complete loss in larvicidal activity was observed for the C199A/P193A double mutant, whereas a small reduction in toxicity was shown for the C199A/P194A and C199A/P196A mutants. Increasing the flexibility of the alpha4 alpha5 loop through C199A/P193G, C199A/P194G/P196A, C199A/P194A/P196G, and C199A/P194G/P196G mutations significantly decreased the larvicidal activity. Similar to the wild-type protoxin, all mutant toxins were structurally stable upon solubilisation and trypsin activation in carbonate buffer, pH 9.0. These findings are the first biological evidence for a structural function in larvicidal activity of the unique disulphide bridge as well as the proline-rich motif within the alpha4-alpha5 loop of the Cry4Aa toxin. PMID- 15796916 TI - Site-directed mutagenesis and molecular modelling studies show the role of Asp82 and cysteines in rat acylase 1, a member of the M20 family. AB - Acylase 1 from rat kidney catalyzes the hydrolysis of acyl-amino acids. Sequence alignment has shown that this enzyme belongs to the metalloprotein family M20. Site-directed mutagenesis experiments led to the identification of one functionally important amino acid residue located near one of the zinc coordinating residues, which play a critical role in the enzymatic activity. The D82N- and D82E-substituted forms showed no significant activity and very low activity, respectively, along with a loss of zinc coordination. Molecular modelling investigations indicated a putative role of D82 in ensuring a proper protonation of catalytic histidine. In addition, none of the five cysteine residues present in the rat kidney acylase 1 sequence seemed involved in the catalytic process: the loss of activity induced by the C294A substitution was probably due to a conformational change in the 3D structure. PMID- 15796917 TI - Toll-like receptor 2 is expressed on the intestinal M cells in swine. AB - The Toll-like receptor (TLR) 2 binds a wide variety of microbial cell wall components. In this study, we investigated the expression pattern of TLR2 in adult swine gut-associated lymphoid tissues using real-time quantitative PCR, Western blotting, immunohistochemistry, and flow cytometric analysis. The mRNA for TLR2 was preferentially expressed in the mesenteric lymph nodes (MLNs) and Peyer's patches (Pps) of adult swine. Expression in these two tissues was approximately 15- and 9-fold higher than that of spleen, respectively. Western blotting further confirmed that the TLR2 protein was highly expressed in the MLNs and Pps. Interestingly, TLR2-expressing cells were found not only in immune cells, such as T cells and B cells, but also in membranous (M) cells. In addition, double immunostaining for TLR2 and cytokeratin 18 revealed that TLR2 was strongly expressed not only in the cytoplasm but also in the apical membrane of the pocket-like M cells. These results indicate that TLR2 on the MLNs and Pps enable the host defense to respond to a variety of cell wall components. Furthermore, the potential function of TLR2 as a pattern recognition receptor and its cellular distribution suggest that TLR2 plays an important role in ligand specific transcytosis and transport in M cells. PMID- 15796918 TI - Silencing the cardiac potassium channel Kv4.3 by RNA interference in a CHO expression system. AB - RNA interference (RNAi) is a powerful technique for gene silencing, in which the downregulation of mRNA is triggered by short RNAs complementary to a target mRNA sequence, with consequent reduction of the encoded protein. The aim of this study was to test the effects of silencing the expression of the cardiac potassium channel Kv4.3 in a heterologous expression system, in order to investigate the effect of RNAi on channel properties. A Chinese hamster ovary cell line stably expressing Kv4.3 and the accessory beta-subunit KChIP2 was transfected with small interfering RNAs (siRNAs) targeting Kv4.3. Effects of RNAi were monitored at the mRNA, protein, and functional levels. Real-time PCR and immunofluorescence staining revealed significant reduction of Kv4.3 mRNA and protein expression. These results were confirmed by functional patch-clamp measurements of the transient outward current (I(to)) which was reduced up to 80% by RNAi. We conclude that the use of siRNAs reagents for post-transcriptional gene silencing is a new effective method for the reduction of the expression and function of different ionic channels which may be adapted for studying their role also in native cells. PMID- 15796919 TI - Generation of a novel anti-geldanamycin antibody. AB - Geldanamycin (GA) and herbimycin A are benzoquinone ansamycins (BAs) that inhibit the molecular chaperone HSP90. The central role of HSP90 in maintaining the conformation, stability, and function of key oncogenic proteins involved in signal transduction pathways renders BAs attractive candidates for clinical development. Two GA derivatives, 17-allylamino-17-demethoxygeldanamycin and 17 demethoxy-17-N,N-dimethylaminoethylamino-geldanamycin are currently evaluated in clinical trials. The present study demonstrates generation of a polyclonal antibody elicited against GA that was conjugated to keyhole limpet hemocyanin via its 17 position. The anti-GA antibody recognizes GA as well as other BAs, suggesting its possible application for monitoring plasma levels of GA derivatives. The specificity of the antibody towards BAs is demonstrated by its inability to recognize radicicol, an HSP90 inhibitor not related to BAs. This antibody thus presents a novel research tool as well as a possible alternative approach for monitoring drug levels in patients. PMID- 15796920 TI - Quinone reduction by Rhodothermus marinus succinate:menaquinone oxidoreductase is not stimulated by the membrane potential. AB - Succinate:quinone oxidoreductase (SQR), a di-haem enzyme purified from Rhodothermus marinus, reveals an HQNO-sensitive succinate:quinone oxidoreductase activity with several menaquinone analogues as electron acceptors that decreases with lowering the redox midpoint potential of the quinones. A turnover with the low-potential 2,3-dimethyl-1,4-naphthoquinone that is the closest analogue of menaquinone, although low, can be detected in liposome-reconstituted SQR. Reduction of the quinone is not stimulated by an imposed K+-diffusion membrane potential of a physiological sign (positive inside the vesicles). Nor does the imposed membrane potential increase the reduction level of the haems in R. marinus SQR poised with the succinate/fumarate redox couple. The data do not support a widely discussed hypothesis on the electrogenic transmembrane electron transfer from succinate to menaquinone catalysed by di-haem SQRs. The role of the membrane potential in regulation of the SQR activity is discussed. PMID- 15796921 TI - Inflammatory gene expression in Coxsackievirus B-4-infected human islets of Langerhans. AB - The event that triggers the autoimmune destruction of insulin-producing beta cells in type 1 diabetes mellitus (T1DM) is still unknown. Enterovirus, especially Coxsackievirus, infections have long been associated with this disease. Cytokines and chemokines induced by an enterovirus infection may act to trigger the autoimmune reactions that produce T1DM. Gene expression was examined in isolated human islets infected with a Coxsackievirus-B4 (CBV-4) strain causing lytic infection (V89-4557) and in islets infected with a CBV-4 strain establishing persistent infection (VD2921). Microarray analysis indicated that infection with the CBV-4 strains resulted in specific induction of a number of inflammatory genes, including IL-1beta, IL-6, IL-8, MCP-1, and RANTES. Importantly, the inflammatory genes induced by the CBV-4 infections differed in the two strains, with more cytokines being induced by the non-lytic CBV-4 strain than by the lytic strain. These cytokines and chemokines have the potential to rapidly induce inflammatory reactions when expressed in vivo and could contribute to the autoimmune reactions associated with the development of T1DM. PMID- 15796923 TI - A synthetic peptide forms voltage-gated porin-like ion channels in lipid bilayer membranes. AB - Design of simple protein structures represents the essential first step toward novel macromolecules and understanding the basic principles of protein folding. Our work focuses on the ion channel formation and structure of peptides having a repeated pattern of glycine residues. Investigation of the ion channel properties of a glycine repeat peptide, VSLGLSIGFSVGVSIGWSFGRSRG revealed the formation of porin-like high conductance, multimeric, non-selective voltage-gated channels in phospholipid bilayer membranes. ATR-IR and CD spectroscopic studies showed an anti-parallel beta sheet structure in membranes. The formation of porin-like ion channels by a beta sheet peptide suggests spontaneous assembly into a beta barrel structure through oligomerization as in pore forming bacterial toxins. The present work is the first example of a short synthetic peptide mimicking the pore characteristics of a complex beta barrel protein and demonstrates that smaller peptides are capable of mimicking the complex functional properties of natural ion channels. This will have implications in understanding the folding of beta sheet proteins in membranes, the mechanism of two state voltage gating, and the role of glycine residues in beta barrel proteins. PMID- 15796922 TI - The long-acting glucagon-like peptide-1 analogue, liraglutide, inhibits beta-cell apoptosis in vitro. AB - We here show that GLP-1 and the long-acting GLP-1 analogue, liraglutide, interfere with diabetes-associated apoptotic processes in the beta-cell. Studies using primary neonatal rat islets showed that native GLP-1 and liraglutide inhibited both cytokine- and free fatty acid-induced apoptosis in a dose dependent manner. The anti-apoptotic effect of liraglutide was mediated by the GLP-1 receptor as the specific GLP-1 receptor antagonist, exendin(9-39), blocked the effects. The adenylate cyclase activator, forskolin, had an anti-apoptotic effect similar to those of GLP-1 and liraglutide indicating that the effect was cAMP-mediated. Blocking the PI3 kinase pathway using wortmannin but not the MAP kinase pathways by PD98059 inhibited the effects of liraglutide. In conclusion, GLP-1 receptor activation has anti-apoptotic effect on both cytokine, and free fatty acid-induced apoptosis in primary islet-cells, thus suggesting that the long-acting GLP-1 analogue, liraglutide, may be useful for retaining beta-cell mass in both type 1 and type 2 diabetic patients. PMID- 15796924 TI - Preparation and characterization of a chimeric zebrafish-human neuroglobin engineered by module substitution. AB - Neuroglobin (Ngb) is a recently discovered vertebrate heme protein that can reversibly bind oxygen that is expressed in the brain. Zebrafish and human Ngb share about 50% amino acid sequence identity. These Ngb proteins consist of four compact protein structural unit "modules" referred to as M1-M4. In the present study, we investigated the effects of module substitution on the properties of Ngb. Specifically, we prepared and characterized a chimeric ZHZZ Ngb in which the heme-binding module M2 of zebrafish Ngb was replaced by the comparable human Ngb module. Our results showed that the chimeric ZHZZ was stable and formed almost the identical heme-environmental and alpha-helical structure as the human and zebrafish Ngb proteins, suggesting that the structure of Ngb has been evolutionarily conserved. PMID- 15796926 TI - Localization and function of three monothiol glutaredoxins in Schizosaccharomyces pombe. AB - The fission yeast Schizosaccharomyces pombe contains two dithiol glutaredoxins (Grx1 and Grx2) and genes for three putative monothiol glutaredoxins (grx3, 4, and 5). We investigated the expression, sub-cellular localization, and functions of the three monothiol glutaredoxins. Fluorescence microscopy revealed that Grx3 is targeted to nuclear rim and endoplasmic reticulum, Grx4 primarily to the nucleus, and Grx5 to mitochondria. Null mutation of grx3 did not significantly affect growth and resistance against various oxidants, whereas grx5 mutation caused slow growth and sensitivity toward oxidants such as hydrogen peroxide, paraquat, and diamide. The grx2grx5 double mutation, deficient in all mitochondrial glutaredoxins, caused further retardation in growth and severe sensitivity toward all the oxidants tested. The grx4 mutation was not viable, suggesting a critical role of Grx4 for the physiology of S. pombe. Overproduction of Grx3 and Grx5, but not the truncated form of Grx5 without mitochondrial target sequence, severely retarded growth as Grx2 did, supporting the idea that Grx2, 3, and 5 are targeted to organellar compartments. Our results propose a distinct role for each glutaredoxin to maintain thiol redox balance, and hence the growth and stress resistance, of the fission yeast. PMID- 15796925 TI - The newly discovered insect order Mantophasmatodea contains a novel member of the adipokinetic hormone family of peptides. AB - A novel member of the AKH/RPCH family of peptides has been identified from the corpus cardiacum of an, as yet, unidentified species of the newly discovered insect order Mantophasmatodea from Namibia. The primary sequence of the peptide, which is denoted Manto-CC, was deduced from multiple MS(N) electrospray mass data to be an octapeptide: pGlu-Val-Asn-Phe-Ser-Pro-Gly-Trp amide. Synthetic Manto-CC co-elutes on reversed-phase HPLC with the natural peptide from the gland of the insect. Interestingly, Manto-CC is structurally very closely related (only one point mutation) to the AKH/RPCH peptides previously identified in mostly more basal insect taxa (Odonata, Blattodea, and Ensifera) and in Crustacea, the sister group of insects, whereas larger structural differences occur with peptides from Mantodea and Phasmatodea, which are thought to be close relatives of Mantophasmatodea. Functionally, Manto-CC may be employed to activate glycogen phosphorylase to mobilize carbohydrates. PMID- 15796927 TI - Involvement in K+ access of Leu318 at the extracellular domain flanking M3 and M4 of the Na+,K+-ATPase alpha-subunit. AB - The effect of point mutation in the sequence 316TWLE319, which occurs in the extracellular loop flanking the third (M3) and the fourth (M4) transmembrane segment (L3/4) of the Na+,K+-ATPase alpha-subunit, was examined. Mutation of Glu319 to Asp yielded an enzyme with full activity, whereas substituting Glu319 to Ala resulted in a severe loss of activity. A negative charge was introduced along the sequence, one residue at a time, from Thr316 to Leu318 (by E-scanning) in the mutant construct with Glu319 already mutated to Gln. The activity that had been reduced to 60% by the mutation of Glu319 to Gln was restored upon the introduction of a negative charge by E-scanning. When Leu318 was replaced by Glu in a series of scanning experiments, the K+ sensitivity of the ATPase activity was lowered. The lowering of K+ sensitivity was further demonstrated when a mutation of Leu318 to Glu was introduced into the wild-type enzyme. Furthermore, mutants with Leu318 to Gln, Arg, and Phe displayed lower K+ sensitivity similar to that of Leu318 to Glu mutant. Leu318 may be in access path for K+, and any substitution at this position may interfere with access of K+ from outside the cell. PMID- 15796928 TI - Compartmentalization of NO signaling cascade in skeletal muscles. AB - Skeletal muscle functions regulated by NO are now firmly established. However, the literature on the compartmentalization of NO signaling in myocytes is highly controversial. To address this issue, we examined localization of enzymes engaged in L-arginine-NO-cGMP signaling in the rat quadriceps muscle. Employing immunocytochemical labeling complemented with tyramide signal amplification and electron microscopy, we found NO synthase expressed not only in the sarcolemma, but also along contractile fibers, in the sarcoplasmic reticulum and mitochondria. The expression pattern of NO synthase in myocytes showed striking parallels with the enzymes engaged in L-arginine-NO-cGMP signaling (arginase, phosphodiesterase, and soluble guanylyl cyclase). Our findings are indicative of an autocrine fashion of NO signaling in skeletal muscles at both cellular and subcellular levels, and challenge the notion that the NO generation is restricted to the sarcolemma. PMID- 15796929 TI - A new function of isonitrile as an inhibitor of the Pdr5p multidrug ABC transporter in Saccharomyces cerevisiae. AB - Pdr5p in Saccharomyces cerevisiae is a functional homologue of mammalian P glycoprotein implicated in multidrug resistance (MDR). In order to obtain useful inhibitors to overcome MDR in clinical tumors, screening of Pdr5p inhibitors has been carried out. We isolated a fungal strain producing Pdr5p inhibitors using our original assay system, and it was classified as Trichoderma sp. P24-3. The purified inhibitor was identified as isonitrile, 3-(3'-isocyano-cyclopent-2' enylidene)-propionic acid, a compound whose carboxyl residue is essential for the inhibitory activity. A non-toxic concentration of the isonitrile (41.5 microg/ml, 255 microM) inhibited Pdr5p-mediated efflux of cycloheximide or cerulenin in Pdr5p-overexpressing cells. In addition, addition of the isonitrile led to accumulation of rhodamine 6G, a substrate of Pdr5p, in the Pdr5p-overexpressing cells. The inhibitory profiles of the isonitrile against S1360 mutants (S1360A and S1360F) of Pdr5p were different from those of FK506 and enniatin. The isonitrile did not influence PDR5 gene expression and the amount of Pdr5 protein, nor did it inhibit the function of Snq2p, a homologue of Pdr5p. Interestingly, the isonitrile inhibited the function of Cdr1p and Cdr2p, Pdr5p homologues in pathogenic yeast Candida albicans. Thus, it was found that the isonitrile shows a different inhibitory spectrum from that of FK506 and enniatin as a potent inhibitor for Pdr5p, Cdr1p, and Cdr2p. PMID- 15796930 TI - A touch of richness. PMID- 15796931 TI - The intensely personal and sometimes unexpected adventures of gastric bypass surgery. PMID- 15796932 TI - A phoenix from the fire: reviewing the rebirth of C-reactive protein research. PMID- 15796933 TI - Esophageal cancer: is there hope? PMID- 15796934 TI - I cannot smile or wink anymore: facial nerve weakness after acoustic neuroma surgery. PMID- 15796935 TI - Thromboembolic prophylaxis in orthopedic and general surgery. PMID- 15796936 TI - Chest wall reconstruction. PMID- 15796937 TI - On the other hand: an update on carpal tunnel syndrome. PMID- 15796938 TI - Current status of stents for arterial occlusive disease and endovascular aortic aneurysm repair. PMID- 15796939 TI - Historical perspective and current management of colonic and intraperitoneal rectal trauma. PMID- 15796940 TI - A case of mistaken identity: giant cystic pheochromocytoma. AB - A 39-year-old Marshall Islands woman was referred for evaluation of an abdominal mass. Medical history was significant only for pulmonary tuberculosis and scrofula. The patient denied a personal or family history of pancreatic or endocrine disease. Physical examination revealed normal vital signs and a 12-cm mildly tender left upper quadrant mass. A computed tomography scan revealed an 11 cm cystic mass contiguous with the distal pancreas and closely associated with the hilum of the spleen and the left kidney. Based on these findings and the patient's history, a cystic neoplasm of the pancreas was suspected, and she was subsequently taken to the operating room for exploration. Intraoperatively, the patient became markedly hypertensive with manipulation of the mass, requiring intravenous nitroprusside. Histopathological evaluation confirmed the diagnosis of cystic pheochromocytoma (PCC). The patient's postoperative course was uncomplicated. Cystic PCCs may not present with the classic prodromal symptoms associated with solid PCCs. This case represents the complex and unsuspected presentation of an extremely rare functional cystic neoplasm. A high index of suspicion for cystic PCC is necessary when confronted with cystic lesions in the vicinity of the adrenal glands. Failure to recognize cystic PCC before resection may lead to uncontrollable hypertension in the operating room, with potentially serious consequences. PMID- 15796941 TI - Are Army-trained surgeons satisfied with their residencies? AB - OBJECTIVE: To evaluate the perceptions of training adequacy among surgeons educated in Army general surgical residencies as a tool for surgical program directors and students considering a military surgical career. DESIGN: A questionnaire was sent to all general surgeons practicing in the Army during years 1999-2003 (n = 182). In addition to providing basic demographic information, subjects rated their perceptions of training experience in 13 areas on a 5-point Likert scale (1 = very dissatisfied, 2 = somewhat dissatisfied, 3 = neither satisfied nor dissatisfied, 4 = somewhat satisfied, 5 = very satisfied). Respondents were split into 3 groups based on graduation year (1968-1992, 1993 1998, 1999-2003) and thereby roughly on status of military obligation at the time of survey. Scores were compared with analysis of variance. RESULTS: A total of 96 (52.7%) questionnaires were returned, 84 of which were included in this study. The average score for all graduation groups and satisfaction areas was 4.37 +/- 0.91. No differences occurred among the 3 graduation groups in any of the 13 satisfaction areas evaluated, except for pediatric surgery experience, where the most recent graduates rated their satisfaction lower than the other 2 groups (1968-1992, 4.00 +/- 0.61; 1993-1998, 3.96 +/- 1.14; 1999-2003, 3.21 +/- 1.27, p < 0.05). With respect to comparison among the 13 satisfaction areas, several areas of note are present. Satisfaction with training in care for basic surgical problems and the ability to make correct decisions are both higher than 5 other areas (p < 0.01). By contrast, satisfaction with number of cases performed, research training, and pediatric surgery training are lower than at least 3 of the other 12 areas (p < 0.01). CONCLUSION: Army trained general surgeons, from the most distant to recent graduates, are satisfied with their residencies. Lower satisfaction scores in the areas of number of cases performed, research experience, and pediatric surgery training do, however, highlight aspects for continued research and improvement. PMID- 15796942 TI - Bilious vomiting and abdominal pain in an aesthenic man. PMID- 15796943 TI - Factors affecting the professional image of physicians. AB - OBJECTIVE: The purpose of this study was to use an experimental design to determine the effect of selected aspects of dress on the professional image of physicians in an adult outpatient setting by those who use the health care services the most (55+), the least (18 to 22), and other physicians on whom we depend for referrals. METHODS: Ten slides of physicians representing a variety of ages; ethnic and religious affiliations were shown to 3 populations across the United States and Canada. Set 1 (N = 216) was shown to undergraduates and rated on 10 attributes of professional image. Two slides were altered and retested in a rotated order (Set 2). Set 1 was then tested with groups of elderly and surgeons (N = 277). RESULTS: Significant differences were found between Set 1 and Set 2 for the altered physician slides. There was high correlation of professionalism with the identified attributes for all 3 rater populations. The nonverbal attributes chosen for this study did accurately assess "professionalism." Gender of the rater or physician did not have any effect. Although the most "professional" surgeon was the oldest in the study, the second most "professional" was the youngest. CONCLUSIONS: It is clear that a surgeon's image is a mirror of competence, trust, expertise, and compassion. The variables described in this study may easily be implemented to reflect a more positive professional image with our peers and patients than simply adding a white coat. No one goes to see a surgeon to establish a casual relationship, and because it is difficult to counteract initial impressions, attention to these variables is important. PMID- 15796944 TI - Finding the best abdominal closure: an evidence-based review of the literature. AB - BACKGROUND: Despite advances in surgical technique and materials, abdominal fascial closure has remained a procedure that often reflects a surgeon's personal preference with a reliance on tradition and anecdotal experience. The value of a particular abdominal fascial closure technique may be measured by the incidence of early and late wound complications, and the best abdominal closure technique should be fast, easy, and cost-effective, while preventing both early and late complications. This study addresses the closure of the vertical midline laparotomy incision. DATA SOURCES: A MEDLINE (National Library of Medicine, Bethesda, Maryland) search was performed. All articles related to abdominal fascia closure published from 1966 to 2003 were included in the review. CONCLUSIONS: Careful analysis of the current surgical literature, including 4 recently published meta-analyses, indicates that a consistent conclusion can be made regarding an optimal technique. That technique involves mass closure, incorporating all of the layers of the abdominal wall (except skin) as 1 structure, in a simple running technique, using #1 or #2 absorbable monofilament suture material with a suture length to wound length ratio of 4 to 1. PMID- 15796945 TI - New anatomic repair of midline abdominal wall incisions extending to suprapubic region. AB - This article describes a new technique to close infraumbilical midline abdominal wall incisions extending to the suprapubic region. This repair is technically easy to perform, saves time, reduces the risk of bowel and bladder injury and takes into consideration the anatomical landmarks of the peritoneum, the rectus sheath and the arcuate line. Abdominal wall closure with this repair is especially useful in obese patients. Furthermore, with this repair the pelvic cavity is lined with smooth peritoneum, which may reduce postoperative adhesions in the pelvis. Technically easy to perform and saves time Very useful technique for obese patients Reduced risk of bowel and bladder injury Anatomical in nature Potentially minimizes postoperative adhesions. PMID- 15796946 TI - Residency reform: anticipated effects of ACGME guidelines on general surgery and internal medicine residency programs. AB - INTRODUCTION: The Accreditation Council for Graduate Medical Education (ACGME) has recently amended guidelines for resident work environment. This study was conducted to evaluate opinions of program directors regarding the impact of the changes on residents and residency programs. METHODS: General surgery and internal medicine program directors were sent a 19-question survey. Questions were asked regarding anticipated effects on patient safety, resident well-being, education, medical errors, implementation costs, and methods needed for compliance. Data were analyzed using the chi-square test, the Mann-Whitney method, and the independent samples t-test where appropriate. RESULTS: Responses were received from 153 surgery program directors and 126 medicine program directors. Differences noted were hours worked (surgery 84.2 hours vs medicine 68.7 hours, p < 0.0005), current compliance (49% vs 73%, p < 0.0005), and allowance of internal (13% vs 54%, p < 0.0005) and external (24% vs 58%, p < 0.0005) moonlighting. CONCLUSIONS: Program directors anticipate improved resident safety and well-being. However, education, continuity of care, and board certification success are not expected to improve. Increased cost to institutions is anticipated. Surgery program directors feel medical errors will not decrease; medicine program directors are neutral. To facilitate compliance, surgery program directors anticipate employing physicians' assistants and technology, whereas medicine program directors may implement night float. Neither surgery nor medicine program directors expects increased quantity or quality of applicants. Program directors agree resident work hour reform is essential; however, varied methodology and outcomes are expected. PMID- 15796947 TI - Asymptote. PMID- 15796948 TI - Maintaining personal health while providing public care: a surgical resident's experience. PMID- 15796950 TI - Staying healthy during surgery residency. PMID- 15796949 TI - My Bushido's code to a healthy surgical lifestyle. PMID- 15796951 TI - Classic groin hernia repair...lest we forget. AB - Currently, operative repair of inguinal hernia is most often performed using one of the open mesh procedures or laparoscopic techniques. These newer approaches minimize anatomical dissection critical to the time-honored traditional hernia surgery described by Bassini, Halsted, McVay, Laroque, Shouldice, and other early pioneers. The familiarity with groin anatomy and the technical skill gained in performing these operations is currently missing from present-day surgical residency training. This article reviews 5 classic hernia operations described by the surgeons whose name they bear, with a view toward better understanding the authors' techniques and philosophies. Each of these operations, though considered by some as of historical interest only, offers today's surgeon reliable alternatives when the simple application of mesh by open or laparoscopic technique is inappropriate. (Curr Surg 62:249-252. Published by Elsevier Inc. on behalf of the Association of Program Directors in Surgery.). PMID- 15796953 TI - Impact of menopausal status on the behavior of pancreatic cystic neoplasms in women. AB - PURPOSE: Pancreatic cystic neoplasms have predilections for occurring in young women, for containing "ovarian-like" stroma, and for expressing estrogen and progesterone receptors; these factors suggest a potentially important influence of the hormonal milieu on the biology of these tumors. Therefore, we examined the impact of menopausal status on the clinical features of pancreatic cystic neoplasms in women. METHODS: Seventy-six women with pancreatic cystic neoplasms treated at our institution from 1992 to 2003 were classified into 2 groups according to menopausal status based on the U.S. average menopausal age of 51 years: group I (premenopausal, ages 22-50; n = 36) and group II (postmenopausal, ages 51-80; n = 40). Chi-square and the 2-tailed t-tests compared categorical and continuous variables, respectively. Kaplan-Meier survival estimates were determined and compared with the log rank test. RESULTS: Abdominal pain at presentation occurred more commonly among group I than among group II patients (78% vs 48%, p < 0.05). Solid pseudopapillary tumors were more prevalent among group I than among group II patients (21% vs 3%, p = 0.02). CONCLUSIONS: The clinicopathologic features of pancreatic cystic neoplasms in premenopausal women are not significantly different from those in postmenopausal women. Menopausal status should not bias diagnostic and treatment algorithms for women with these neoplasms. PMID- 15796952 TI - Core biopsy diagnosis of ductal carcinoma in situ: an indication for sentinel lymph node biopsy. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) is a minimally invasive, accurate method of evaluating axillary lymph nodes in patients with invasive cancer. The technique has also been applied successfully in patients with ductal carcinoma in situ (DCIS). The purpose of this study was to review our experience performing SLNB in patients with a biopsy diagnosis of DCIS. METHODS: A prospective study of consecutive patients seen at our institution from August 2001 to April 2004 with a biopsy diagnosis of DCIS was undertaken. Demographic data, biopsy method, final pathology, and surgical treatment were recorded. Patients undergoing SLNB were identified, and pathologic results were noted. RESULTS: Eighty-five patients with a biopsy diagnosis of DCIS were treated. Fifty-five (64.7%) had their diagnosis made by excisional biopsy, and 30 (35.3%) by core biopsy. Forty-four (51.7%) patients underwent SLNB as part of their definitive surgical procedure, and an SLN was successfully identified in 41 (93.2%). Nine (22.0%) patients who underwent successful SLNB had a positive SLN, 2 by hematoxylin and eosin (H&E) staining and 7 by immunohistochemical (IHC) staining for cytokeratin. Both patients with H&E-positive SLN were ultimately found to have invasive disease in their primary lesion. Final pathologic assessment of all primary lesions revealed invasive carcinoma in 7, 6 of whom had their diagnosis made by core biopsy. Overall, 20.0% of patients with a core biopsy diagnosis of DCIS were upstaged to invasive disease. Whether the lesion was palpable, grade and the presence or absence of necrosis were not significantly different in patients ultimately found to have invasive disease versus those who did not. DISCUSSION: Sentinel lymph node biopsy can be performed accurately in patients with a biopsy diagnosis of DCIS. The rate of axillary disease in patients with pure, completely resected DCIS is low; therefore, SLNB is not indicated in all patients with this biopsy diagnosis. Because of a high rate of invasive disease on the final pathology of patients with DCIS diagnosed by core biopsy, these patients should be offered SLNB. PMID- 15796954 TI - Surgical robotics. PMID- 15796955 TI - The legacy of Allen Oldfather Whipple. PMID- 15796956 TI - Targeting plasma cells improves detection of cytogenetic aberrations in multiple myeloma: phenotype/genotype fluorescence in situ hybridization. AB - Standard fluorescence in situ hybridization (FISH) easily detects nonrandom karyotypic abnormalities in multiple myeloma (MM) at disease presentation, when tumor burden is high. In contrast, the detection of residual MM using the standard 200 unselected nonmitotic nuclei FISH approach correlates poorly with residual disease detected by morphology, flow cytometry, immunohistochemistry, or reverse-transcription polymerase chain reaction (RT-PCR). We have used sequential May-Grunwald Giemsa stain to identify plasma cell populations, followed by FISH analyses (target FISH or T-FISH) to detect immunoglobulin heavy-chain gene (IGH) rearrangements, 13q or 17p deletions, or hyperdiploidy. In this study, 115 samples were collected from 100 patients with MM regardless of treatment status. In this proof-of-principle prospective study, T-FISH detected MM in 52 samples (45%), a percentage similar to that obtained by pathology. Disease detection increased from 5.6% with standard FISH to 48% with T-FISH, and cell culture experiments showed that T-FISH consistently detected a clonal abnormality at dilutions of 10(-3). In five patients, T-FISH further identified myelodysplastic associated karyotypic changes restricted to myeloid cells. Our observations suggest that T-FISH identifies cell lineage involvement of cytogenetic abnormalities, improves detection of low-level or residual MM, and may define the coexistence of hematologic karyotypic changes in individual patients. PMID- 15796957 TI - Characterization of quantitative chromosomal abnormalities in renal cell carcinomas by interphase four-color fluorescence in situ hybridization. AB - Renal cell carcinomas (RCC) in adults are histologically heterogeneous solid tumors with specific chromosomal abnormality patterns included in the World Health Organization (WHO) classification. To overcome some of the drawbacks of cytogenetic and comparative genomic hybridization (CGH) analyses, we designed a first-generation cytogenetic diagnostic test using four-color fluorescence in situ hybridization (FISH) on interphase nuclei. We selected 51 bacterial artificial chromosome and P1-derived artificial chromosome clones covering 17 chromosomal regions involved in the abnormalities of the adult RCC histologic subtypes. An initial set of probes allowed the identification of clear-cell RCC, papillary RCC, and other RCC on a single slide. A second test allowed the detection of additional chromosomal abnormalities or aberrations specific to chromophobic RCC and oncocytomas. We tested 25 cases of RCC, and the results were in agreement with those of cytogenetic techniques and/or CGH methods. The techniques appeared to be very sensitive, because small tumoral cell clones that were undetected by other cytogenetic methods were identified with this method. It was concluded that the multicolor FISH test was specific and sensitive, easy to perform, and could be part of the investigation process in RCC. PMID- 15796958 TI - BRCA1 and pancreatic cancer: pedigree findings and their causal relationships. AB - Anecdotal reports and series studies indicate that 5-10% of pancreatic cancer (PC) cases are familial. In addition, PC is associated with a variety of hereditary cancer syndromes. PC appears to be an integral cancer in the hereditary breast-ovarian cancer (HBOC) syndrome, with most notice given to the role of BRCA2. Our purpose is to call attention to BRCA1, which also predisposes to PC. Using data from our familial breast cancer registry, we identified 19 BRCA1/2 families that contain PC affecteds in the pedigrees, 15 with BRCA1 mutations and 4 with BRCA2 mutations. The association between BRCA2 and pancreatic cancer is well established; however, a definite link with pancreatic cancer in families carrying a BRCA1 mutation has been far less studied. Thus, the focus of this report is on 9 of the 15 BRCA1 families, in which PC affecteds were either confirmed carriers of the BRCA1 mutation or were inferred as probable obligate BRCA1 mutation carriers. The numbers are small, but nevertheless illustrate the finding of others of an apparent association between PC and BRCA1 mutation-bearing families. Given the dismal prognosis of PC, with the only current hope for survival being through surgical extirpation of the pancreas prior to metastasis, it is prudent that we realize the potential predisposition toward PC via BRCA1, in the hope of early diagnosis and prevention. PMID- 15796959 TI - Molecular analysis of primary and recurrent giant cell tumors of bone. AB - The status of microsatellite markers located on chromosomes 1p36, 3p25, 5q23, 9p22, 10q23, 10q24, 17p13, and 19q12 was used to determine loss of heterozygosity (LOH) in primary giant cell tumors (GCT) of bone in 12 patients. The cases included primary, locally recurrent, and metastatic GCT; three tumors were classified as malignant GCT, based on their morphological features. Microdissection was performed on 24 paraffin-embedded tissue samples. An average of three separate topographic sites were microdissected from each tumor. Case selection in each instance was based on the availability of paired samples of tumor in primary GCTs and their corresponding recurrences, and the presence of normal tissue. The number of cases studied is too small for statistical studies, and thus the analysis is descriptive. All cases were informative for >80% of the markers used. Both primary GCTs and local recurrences and lung metastases displayed LOH of three or more markers, and intratumoral heterogeneity was frequent. Fractional allelic losses (FAL) were not different in recurrent and nonrecurrent GCT. FAL was greatest (>30%) in the metastatic group of GCT. Allelic losses of 1p, 9q, and 19q regions were frequent in all groups. LOH of 17p (in proximity to the p53 locus) and 9p occurred exclusively in the pulmonary metastases from GCT. LOH of 9q and 19q was present in primary as well as recurrent GCTs and in one malignant GCT. Involvement of 1p (including MYCL) and 9q regions has not been previously reported in GCT of bone. The pattern of LOH evident in the 17 markers used in the present study suggests that GCT with malignant features may follow an evolutionary pathway similar to the usual primary GCT of bone. PMID- 15796960 TI - Genetic analysis of susceptibility to endometrial adenocarcinoma in the BDII rat model. AB - Most cancers are genetically complex and heterogeneous, a serious obstacle to identifying specific genes underlying the disease. If inbred animal models are used, then both the genetic constitution and environmental influences can be carefully controlled. Females of the BDII inbred rat strain are genetically predisposed to endometrial cancer; more than 90% of virgin BDII females will develop endometrial adenocarcinoma (EAC) during their life span. BDII females were crossed to males from inbred strains with low EAC incidence (SPRD or BN). When F(1) males were backcrossed to BDII females to generate N(1) populations of offspring, about one fourth of the female progeny developed EAC. With transmission disequilibrium test analysis, significant association was detected in three chromosomal regions (on RNO1, RNO11, and RNO17) in the SPRD crosses and in the short arm of RNO20 in the BN crosses. It appears that several susceptibility genes with minor but cooperating effects are responsible for the susceptibility. Furthermore, it seems clear from the interstrain crosses not only that the onset of tumors depends on the presence of susceptibility alleles from the EAC-prone BDII strain, but also that tumor development is affected by the contribution of a genetic component derived from the nonsusceptible strains. PMID- 15796961 TI - Allelic loss at 10q26 in osteosarcoma in the region of the BUB3 and FGFR2 genes. AB - Loss of heterozygosity at 10q26 was mapped using microsatellite markers in 20 osteosarcomas. A four-megabase region centered on marker D10S587 was affected by allelic loss in 60 percent of osteosarcomas. The most frequently lost marker was D10S1723. Around 15 known genes are found in this region. The gene immediately adjacent to D10S1723 encodes BUB3, an element of the spindle assembly mitotic checkpoint. Loss of BUB3 function could contribute to chromosomal instability. The fibroblast growth factor receptor 2 (FGFR2) gene is located 2 Mb from the BUB3 gene and has the potential for a role in cancer. Inherited mutations of the FGFR2 gene result in skeletal dysplasias. FGFR2 alterations have also been implicated in gastric cancer. Human genome project data were used to design primers for amplifying FGFR2 in 18 genomic segments and BUB3 in 7 genomic segments. In each case, the segments encompassed coding exons and flanking intron sequences. The primers were used to search for mutations by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP). Several shifted bands were detected in the BUB3 exon 3 fragment. Sequencing resolved the BUB3 exon 3 fragment shifts into polymorphisms in intron 2. No mutations of BUB3 or FGFR2 were detected. It remains possible that BUB3 or FGFR2 hemizygosity alone contributes to osteosarcoma, or that one of the genes is cryptically inactivated by a higher-order modification or mutation outside the coding region. There may also be a yet undiscovered tumor suppressor gene in this region. PMID- 15796962 TI - Methylation status of EXT1 and EXT2 promoters and two mutations of EXT2 in chondrosarcoma. AB - Germline mutation and functional loss of EXT1 or EXT2 are commonly found in multiple osteochondromas and predispose to the development of chondrosarcoma. Mutations of EXT1 and EXT2 have rarely been detected in sporadic secondary chondrosarcomas from osteochondroma; these frequently display loss of heterozygosity at the EXT1 and EXT2 loci, but primary chondrosarcomas typically do not. To evaluate promoter methylation (which is an epigenetic gene silencing mechanism) of EXT1 and EXT2, we performed methylation-specific polymerase chain reaction (PCR) for 20 chondrosarcoma cases (12 primary, 3 secondary to osteochondroma, 2 secondary to enchondromatosis, 2 extraskeletal ordinary, and 1 clear cell) and in five cell lines. In addition, mutation analysis of the EXT1 and EXT2 coding regions was performed using PCR-single-strand conformation polymorphism and sequencing analysis for 12 of the 20 chondrosarcoma cases (8 primary, 1 secondary to enchondromatosis, 1 secondary to osteochondroma, and 2 extraskeletal ordinary) and five cell lines. Promoter methylation of EXT1 and EXT2 was not detected in any of the cases, and both EXT1 and EXT2 were expressed in all cell lines. Two missense mutations in EXT2 (D227E and R299H) were detected among the chondrosarcoma cases. When considering tumor development in primary chondrosarcoma, we should include mutations in EXT2, along with the status of other members of the EXT gene family. PMID- 15796964 TI - Alterations of loci encoding PU.1, BOB1, and OCT2 transcription regulators do not correlate with their suppressed expression in Hodgkin lymphoma. AB - Neoplastic cells of Hodgkin lymphoma (HL) originating from germinal or postgerminal center B cells lose their capacity to transcribe and to express surface immunoglobulins (Ig). This defect correlates with the absence of expression of B-cell-specific transcription regulators, including PU.1, BOB1, and OCT2. These findings suggest that Ig impairment in HL is caused by the defective transcription machinery. The mechanism or mechanisms underlying failure of Hodgkin cells to express PU.1, BOB1, and OCT2 remain unclear. The genes encoding for these three respective transcription factors have been mapped at 11p11.2 (SPI1), 11q23.1 (POU2AF1), and 19q13.2 (POU2F2); these are chromosomes recurrently affected in HL. To check the genomic status of PU.1, BOB1, and OCT2 in HL, we performed metaphase fluorescence in situ hybridization (FISH) analysis of 10 HL cases using locus-specific bacterial artificial chromosome clones. FISH signal pattern was correlated with the ploidy level of each analyzed cell and showed recurrent imbalances of the studied loci. The underrepresentation of one or two analyzed regions was detected in five cases; the remaining five cases showed either random losses, a ploidy-equivalent FISH pattern, or overrepresented signals. Neither a constant loss nor genomic aberration of at least one of these genes could be observed in studied cases. These findings indicate that genomic imbalances or rearrangements are not a cause of PU.1, BOB1, and OCT2 deficiency in cHL and argue for another mechanism underlying this phenomenon. PMID- 15796963 TI - Effects of degenerate oligonucleotide-primed polymerase chain reaction amplification and labeling methods on the sensitivity and specificity of metaphase- and array-based comparative genomic hybridization. AB - Degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) is often applied to small amounts of DNA from microdissected tissues in the analyses of chromosomal copy number with comparative genomic hybridization (CGH). The sensitivity and specificity in CGH analyses largely depend on the unbiased amplification and labeling of probe DNA, and the sensitivity and specificity should be high enough to detect one-copy changes in aneuploid cancer cells when accurate assessment of chromosomal instability is needed. The present study was designed to assess the effects of DOP-PCR and labeling method on the sensitivity of metaphase- and array-based CGHs in the detection of one-copy changes in near tetraploid Kato-III cells. By focusing on several chromosomes whose absolute copy numbers were determined by FISH, we first compared the green-to-red ratio profiles of metaphase- and array-based CGH to the absolute copy numbers using the DNA diluted with varying proportions of lymphocyte DNA, with and without prior DOP-PCR amplification, and found that the amplification process scarcely affected the sensitivity but gave slightly lower specificity. Second, we compared random priming (RP) labeling with nick translation (NT) labeling and found that the RP labeling gave fewer false-positive gains and fewer false-negative losses in the detection of one-copy changes. In array CGH, locus-by-locus concordance between the DNAs with and without DOP-PCR amplification was high (nearly 100%) in the gain of three copies or more and the loss of two copies or more. This suggests that we could pinpoint the candidate genes within large-shift losses-gains that are detected with array CGH in microdissected tissues. PMID- 15796965 TI - Establishment of a cell line from a malignant rhabdoid tumor of the liver lacking the function of two tumor suppressor genes, hSNF5/INI1 and p16. AB - Malignant rhabdoid tumors (MRT) of the liver are rare. A few liver MRT cell lines have been established but none has been characterized in detail. Here we describe a new MRT cell line from the liver, which is designated MP-MRT-AN, and describe it in detail. Immunohistochemical assays detected the expression of vimentin and cytokeratin but they were negative for neurofilament, desmin, alpha-smooth muscle actin, alpha-sarcomeric actin, and smooth muscle myosin heavy chains SM1 and SM2. RT-PCR assays revealed that this cell line did not express smooth muscle myosin heavy chain isoforms or MyoD1. No aberration was identified in 22q by G-banded analysis; however, the hSNF5/INI1 gene, a suppressor gene of MRT that maps to 22q11.2, was homozygously deleted from exons 1 to 5 in this cell line. Furthermore, the expression of another tumor suppressor gene, p16 (CDKN2A), was not detected by RT-PCR. This raises the possibility that the aggressive phenotype of malignant rhabdoid tumors is caused by the loss of two or more tumor suppressor genes. PMID- 15796966 TI - Molecular cytogenetic characteristics of the human hepatocellular carcinoma cell line HCCLM3 with high metastatic potential: comparative genomic hybridization and multiplex fluorescence in situ hybridization. AB - The HCCLM3 cell line was established at the authors' institute from the lung metastatic lesions of BALB/c nude mice bearing human hepatocellular carcinoma (HCC) from the metastatic HCC cell line MHCC97-H. It has been shown to have a high potential for lung metastases and extensive metastases when the cells are inoculated subcutaneously or orthotopically in athymic nude mice. In the present study, the molecular cytogenetic characteristics of this cell line were evaluated with conventional G-banding, comparative genomic hybridization, and multiplex fluorescence in situ hybridization. A hyperdiploid karyotype of 53-58 chromosomes with 10 marker chromosomes was identified. The chromosomal aberrations such as i(X)(q10), der(Y)t(Y;18)(q12;p11), der(3)t(3;20) (p25;q13), der(4)t(4;8)(q31;q22)5, der(9)t(9;13)(p21;q22), der(14)t(14;22)(p13;q13), and der(15) t(15;21)(q11;q22) were described for the first time in human HCC cells. The analysis of this cell line through a combination of molecular cytogenetic techniques provides information on the possible molecular mechanisms involved in the metastatic process of HCC. PMID- 15796967 TI - Chromosome 7 abnormalities in acute megakaryoblastic leukemia associated with Down syndrome. AB - A 2-year-old girl with Down syndrome (DS) developed acute megakaryoblastic leukemia (AMKL) following a transient myeloproliferative disorder (TMD). The blast cells showed an altered karyotype of 47,XX,r(7),+21c. Serial cytogenetic studies during the course of the illness showed rapid stepwise clonal chromosome changes, including a ring chromosome 7, associated with treatment refractoriness. We reviewed 10 published cases of Down syndrome-related AMKL (DS-AMKL) showing chromosome 7 abnormalities and found that these changes do not carry the same prognostic weight as for non-DS children. For DS-AMKL, therefore, other prognostic factors besides clonal cytogenetic changes need to be identified for planning optimal therapy. PMID- 15796968 TI - Inv(1)(p22q25) in chronic myeloproliferative disease: constitutional or clonal defect? AB - Pericentric inversion of chromosome 1 is uncommon in chronic myeloproliferative disease. We report the occurrence of an isolated inv(1)(p22q25) in two patients with BCR/ABL-negative chronic myeloproliferative disease. The inv(1) is an acquired clonal abnormality in one patient and a constitutional defect in the other. To our knowledge, this is the first report on the occurrence of inv(1)(p22q25) in hematolymphoid malignancies. PMID- 15796969 TI - Precursor T-lymphoblastic leukemia with an inv(6)(p21.2q27). PMID- 15796970 TI - Multiple reciprocal translocations in carcinomas. PMID- 15796971 TI - Acquired Robertsonian translocations in leukemia: two more cases. PMID- 15796972 TI - AvrPtoB: a bacterial type III effector that both elicits and suppresses programmed cell death associated with plant immunity. AB - Pseudomonas syringae pv. tomato DC3000 is a model pathogen for studying the molecular basis of plant immunity and disease susceptibility in tomato and Arabidopsis. DC3000 uses a type III secretion system to inject effector proteins into the plant cell. Type III effectors are thought to promote bacterial virulence by suppressing plant defenses and enhancing access to nutrients trapped in the plant cell. The AvrPtoB type III effector elicits immunity-associated programmed cell death (PCD) when expressed in tomato plants carrying the Pto resistance protein. However, in the absence of Pto, AvrPtoB functions to suppress PCD and immunity in tomato. Here, we review current research examining the molecular basis of AvrPtoB-mediated elicitation and suppression of plant PCD. In addition, the "trump model" is proposed to explain how resistance proteins successfully elicit immunity-associated PCD in response to effectors that suppress PCD. PMID- 15796973 TI - Powdery mildew susceptibility and biotrophic infection strategies. AB - Plants are resistant to most potentially pathogenic microbes. This forces plant pathogens to develop sophisticated strategies to overcome basic plant resistance, either by masking intrusion or by suppression of host defences. This is particularly true for fungal pathogens, which establish long lasting interactions with living host tissue, without causing visible damage to invaded cells. The interactions of cereal crops and Arabidopsis with powdery mildew fungi are model systems for understanding host resistance. Currently, these systems are also promoting the understanding of fungal infection by identifying fungal pathogenicity and virulence factors and host target sites. This minireview focuses on recent findings about host susceptibility and the way powdery mildew fungi might induce it. PMID- 15796974 TI - Genotypic characterization of Burkholderia cenocepacia strains by rep-PCR and PCR RFLP of the fliC gene. AB - Thirty-five strains of Burkholderia cenocepacia from clinical and environmental sources were characterized genotypically by repetitive sequence PCR (ERIC- and BOX-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the flagellin gene (fliC). In cluster analysis based on the repetitive PCR profiles the strains were composed of five clusters, of which clusters 1, 2 and 3 were more closely related to each other than to clusters 4 and 5. It has been reported that the majority of Burkholderia cepacia complex strains can be separated into two types on the basis of fliC size (types I and II correspond to 1.4 and 1.0 kb, respectively). When the strains were analysed by PCR of fliC, all strains yielded amplified products of 1.0 kb except for three strains. The latter strains gave PCR products of 0.7 kb (atypical type), which belonged to repetitive PCR cluster 5. These results indicated that the majority of B. cenocepacia strains belonged to flagellin type II. In the RFLP analysis of the large fliC amplicons with HaeIII, 10 patterns were observed indicating remarkable variation. Strains grouping in repetitive PCR cluster 4 had a unique fliC RFLP pattern. The results of repetitive PCR typing and PCR-RFLP analysis of fliC showed a strong correlation. Strains belonging to the repetitive PCR clusters 4 or 5 were distinctly different from other B. cenocepacia strains as shown by PCR-RFLP analysis of the fliC gene and phenotypic assays. PMID- 15796975 TI - DNA array analysis of Candida albicans gene expression in response to adherence to polystyrene. AB - Candidiasis is often initiated by the colonization of inert surfaces. In order to elucidate the mechanisms involved in this adherence process, DNA macroarrays were used to analyze the transcriptome of Candida albicans, the main causative agent of this mycoses, in a simple adherence model using germ tubes produced in polystyrene Petri dishes. Non-adherent germ tubes produced on glass surface were used as a control. Analysis of gene expression displayed 77 genes identified as statistically overexpressed in adherent germ tubes. Among these genes, some encoded enzymes participating in metabolism of lipids (such as LIP6), of proteins (such as SAP1) or of carbohydrates (like PGI1, PMI40 and PSA1. Some of these genes have already been reported as playing a role in pathogenesis of C. albicans. However, functions were unknown for a large part (45.5%) of the overexpressed genes which will be analyzed further in order to define their relationship with adherence. PMID- 15796976 TI - Role of superoxide in the germination of Bacillus anthracis endospores. AB - The spore forming Gram-positive bacterium Bacillus anthracis, the causative agent of anthrax, has achieved notoriety due to its use as a bioterror agent. In the environment, B. anthracis exists as a dormant endospore. Germination of endospores during their internalization within the myeloid phagocyte, and the ability of those endospores to survive exposure to antibacterial killing mechanisms such as superoxide (O(2)*-, is a key initial event in the infective process. We report herein that endospores exposed to fluxes of O(2)*- typically found in stimulated phagocytes had no effect on viability. Further endospores of the Sterne strain of B. anthracis were found to scavenge O(2)*-, which may enhance the ability of the bacterium to survive within the hostile environment of the phagolysosome. Most intriguing was the observation that endospore germination was stimulated by a flux of O(2)*- as low as 1 microM/min. Data presented herein suggest that B. anthracis may co-opt O(2)*- which is produced by stimulated myeloid phagocytes and is an essential element of host immunity, as a necessary step in productive infection of the host. PMID- 15796977 TI - Meiothermus timidus sp. nov., a new slightly thermophilic yellow-pigmented species. AB - Several yellow-pigmented isolates, with optimum growth temperatures between 55 and 60 degrees C, were recovered from hot springs in Central Portugal and the Azores. Phylogenetic analysis of the 16S rDNA showed that these organisms represented a new species of the genus Meiothermus. The new isolates could be distinguished from other strains of the species of the genus Meiothermus by biochemical characteristics and the fatty acid composition because they had very high levels of iso C15:0 and iso C17:0 and very low levels of anteiso C17:0 and iso C16:0. On the basis of the results presented here we propose the name Meiothermus timidus for the new species represented by strains SPS-243(T) (=LMG 22897(T)=CIP 108604(T)), RQ-10 and RQ-12. PMID- 15796978 TI - The Photorhabdus Pir toxins are similar to a developmentally regulated insect protein but show no juvenile hormone esterase activity. AB - The genome of the insect pathogen Photorhabdus luminescens strain TT01 contains numerous genes predicting toxins and proteases. Within the P. luminescens TT01 genome, the products of two loci, plu 4093-plu 4092 and plu 4437-plu 4436, show oral insecticidal activity against both moth and mosquito larvae. The proteins encoded by these loci, here termed 'Photorhabdus insect related' (Pir) proteins A and B, show similarity both to delta-endotoxins from Bacillus thuringiensis (Bts) and a developmentally regulated protein from a beetle, Leptinotarsa decemlineata. The beetle protein has been inferred to possess juvenile hormone esterase (JHE) activity due to its developmentally regulated pattern of expression and the Photorhabdus proteins PirA and PirB have been proposed to be mimics of insect JHEs that can disrupt insect metamorphosis by metabolizing the insect growth regulator juvenile hormone (JH) [Nat. Biotechnol. 21 (2003) 1307-1313]. Here we confirm that, when injected together, PirA and PirB from two different Photorhabdus strains have insecticidal activity against caterpillars of the moth Galleria mellonella but show no oral activity against a second moth species Manduca sexta. Direct measurement of JHE activity, however, shows that the Pir proteins are not able to metabolise JH. These data show that the Pir proteins have no JHE activity, as suggested, but leave the mode of action of these interesting proteins uncertain. PMID- 15796979 TI - DNA methylation modulates Salmonella enterica serovar Typhimurium virulence in Caenorhabditis elegans. AB - Salmonella enterica serovar Typhimurium was previously shown to be virulent in Caenorhabditis elegans. Here we demonstrate that DNA adenine methyltransferase (DAM) modulates Salmonella virulence in the nematode, as it does in mice. After 5 days of continual exposure to bacteria, twice as many worms died when exposed to the wild-type than the dam-mutant strain of Salmonella. Similar trends in virulence were observed when worms were exposed to Salmonella strains for 5 h and transferred to the avirulent Escherichia coli OP50. While a 10-fold attenuation was observed in the absence of DAM, the dam-strain was still able to infect and persist in the host worm. Our results further support the use of C. elegans as an accessible and readily studied animal model of bacterial pathogenesis. However, our results suggest that crucial host responses differ between the murine and nematode models. Additionally, we carried out preliminary liquid culture based experiments with the long term goal of developing high throughput animal based screens of DAM inhibitors. PMID- 15796980 TI - Allophanate hydrolase of Oleomonas sagaranensis involved in an ATP-dependent degradation pathway specific to urea. AB - The first prokaryotic urea carboxylase has previously been purified and characterized from Oleomonas sagaranensis. As the results indicated the presence of an ATP-dependent urea degradation pathway in Bacteria, the characterization of the second component of this pathway, allophanate hydrolase, was carried out. The gene encoding allophanate hydrolase was found adjacent to the urea carboxylase gene. The purified, recombinant enzyme exhibited ammonia-generating activity towards allophanate, and, together with urea carboxylase, efficiently produced ammonia from urea in an ATP-dependent manner. The substrate specificity of the enzyme was strict, and analogs of allophanate were not hydrolyzed. Moreover, although the urea carboxylase exhibited carboxylase activity towards urea, acetamide, and formamide, ammonia-releasing activity of the two enzymes combined was detected only towards urea, indicating that the pathway was specific for urea degradation. PMID- 15796981 TI - A hyperactive, Ca2+-dependent antifreeze protein in an Antarctic bacterium. AB - In cold climates, some plants and bacteria that cannot avoid freezing use antifreeze proteins (AFPs) to lessen the destructive effects of ice recrystallization. These AFPs have weak freezing point depression activity, perhaps to avoid sudden, uncontrolled growth of ice. Here, we report on an uncharacteristically powerful bacterial AFP found in an Antarctic strain of the bacterium, Marinomonas primoryensis. It is Ca(2+)-dependent, shows evidence of cooperativity, and can produce over 2 degrees C of freezing point depression. Unlike most AFPs, it does not produce obvious crystal faceting during thermal hysteresis. This AFP might be capable of imparting freezing avoidance to M. primoryensis in ice-covered Antarctic lakes. A hyperactive bacterial AFP has not previously been reported. PMID- 15796982 TI - Construction and use of an stx1 transcriptional fusion to gfp. AB - Shiga toxins (Stxs), also termed Vero toxins, are cytotoxic ribosome inactivating proteins that are produced by a number of gastrointestinal pathogens and that contribute to the severity of the associated diseases. In this work, we constructed and validated a transcriptional fusion of the stx1AB promoter to the gfp reporter gene. The cloned promoter region encompasses both the proximal and the distal promoter regions of stx1AB, mediating control by the host's iron responsive Fur repressor and the Stx prophage's Q antiterminator protein, respectively. The probe was validated by demonstrating its responsiveness towards mitomycin C and EDTA, and the contribution of host and phage encoded factors could be separated by studying stx1 expression in either wild-type or isogenic lysogenic cells. Moreover, stx1AB expressing populations could be visualized by flow cytometry. The potential use of such a probe for non-destructive online detection of stx1AB expression and visualization of stx1AB expressing populations is further discussed. PMID- 15796983 TI - Restriction site polymorphisms in the genes encoding new members of group 3 outer membrane protein family of Brucella spp. AB - Thirty-seven Brucella reference and field strains representing all the species and their biovars were analysed by PCR-RFLP to determine the degree of variation in the genes encoding the new members of group 3 outer membrane protein (Omp) family. Analysis of the omp22 and omp25c/omp25d genes indicated that the restriction patterns were identical for all species and biovars with all restriction enzymes tested, except for Brucella ovis that showed a short 30 bp deletion close to omp22 gene, and for B. abortus biovar 6 and B. ovis that lacked a DdeI site and a HinfI site, respectively, in the omp25c/omp25d genes. Analysis of PCR products of the omp31b gene digested with 20 restriction enzymes revealed that this gene has a greater level of DNA polymorphism than the other genes encoding the new members of group 3 Omp family. A deletion of 232bp was detected in fourteen B melitensis strains from different hosts and from different geographic origins, confirming that this feature is indeed a hallmark of B. melitensis. PCR-RFLP analysis of omp31b with DdeI allowed us to identify species specific markers for B. abortus, B. melitensis, and B. ovis. Finally, by PCR analysis, Southern blot hybridization and DNA sequencing we showed that a large deletion of 15 kb, comprising the entire omp25b gene and 21 more genes, is present in all B. ovis strains, thus confirming previous observations from other authors. PMID- 15796984 TI - A phylogenetic study of commercial Chinese truffles and their allies: taxonomic implications. AB - Phylogenetic relationships of commercial Chinese truffles and their allies were investigated mainly by morphological studies and analyses of the sequences of ITS regions of nuclear ribosomal DNA. Two species, Tuber indicum and T. himalayense, closely related to the European T. melanosporum (the Perigord Truffle), are recognized among commercial Chinese black truffles. Both T. pseudohimalayense and T. sinense should be regarded as synonyms of T. indicum. Tuber species producing excavated ascomata are not monophyletic, suggesting that excavation of ascomata may have evolved more than once, or evolved once during the evolution of truffle species and then was lost once during the evolution of Tuber species. PMID- 15796985 TI - CTX-M and SHV-12 beta-lactamases are the most common extended-spectrum enzymes in clinical isolates of Escherichia coli and Klebsiella pneumoniae collected from 3 university hospitals within Korea. AB - Among the 443 clinical isolates of Escherichia coli and Klebsiella spp. collected between June and November 2003 from 3 university hospitals in Korea, 62 isolates were confirmed as extended-spectrum beta-lactamase (ESBL)- or plasmid-mediated AmpC beta-lactamase-producers by double disk synergy test, PCR and sequencing for beta-lactamase genes. The most frequently identified ESBL gene among E. coli and K. pneumoniae isolates was bla(SHV-12) and bla(CTX-M) (bla(CTX-M-9), bla(CTX-M 14), bla(CTX-M-3), and bla(CTX-M-15)). Four kinds of plasmid-mediated AmpC beta lactamases, ACT-1, CMY-1, CMY-2, and DHA-1, were detected. ESBL production was associated with high levels of resistance to tetracycline, sulfisoxazole, streptomycin, kanamycin, gentamicin and tobramycin when compared to non-ESBL producing isolates. Conclusively, this study suggests that the CTX-M beta lactamases are prevalent and various kinds of plasmid-mediated AmpC enzymes are distributed in clinical isolates of E. coli and Klebsiella spp. in Korea. PMID- 15796986 TI - Orbilia querci sp. nov. and its knob-forming nematophagous anamorph. AB - Orbilia querci, a new nematode-trapping fungus, was found on rotten wood of Quercus sp. in Huai-rou County, Beijing, China. It is characterized by having a tear-shaped spore body in the cylindrical ascospore. Pure culture was obtained from the ascospores. Conidiophores were simple or occasionally branched, bearing a single conidium on the tip. Conidia were spindle-shaped, mostly with 3-septa. Nematodes were captured by means of adhesive stalked knobs. The adhesive knobs were produced frequently on nutritional agar plates even in the absence of challenging nematodes. Its anamorph is placed in Dactylellina and named as D. querci. The sequence divergence of the ITS1 region between the fungus and the other knob-forming species tested was 23.8-33.4%, supporting O. querci as a distinct species. This is the first report of the connection between a knob forming nematophagous hyphomycete and an Orbilia teleomorph. PMID- 15796987 TI - Disruption of a gene encoding glycerol 3-phosphatase from Candida albicans impairs intracellular glycerol accumulation-mediated salt-tolerance. AB - Intracellular glycerol accumulation is critical for Candida albicans to maintain osmolarity, and therefore defects in glycerol homeostasis can have severe effects on the morphogenetic plasticity and pathogenicity of this fungus. The final step of glycerol synthesis involves the dephosphorylation of glycerol 3-phosphate by glycerol 3-phosphatase (GPP1). We have identified a single copy of the GPP orthologous gene (GPP1) in the C. albicans haploid genome, as well as the paralogous gene 2-deoxyglucose-6-phosphate phosphatase (DOG1); both belong to a family of low molecular weight phosphatases. A knockout of the GPP1 gene in C. albicans caused increased susceptibility to high salt concentrations, indicating a deficiency in osmoregulation. Reintroduction of the GPP1 gene complemented the impairment of salt-tolerance in the gpp1/gpp1 mutant. Northern blot analysis showed that the GPP1 gene was strongly responsive to osmotic stress, and its transcriptional expression was positively correlated with intracellular glycerol accumulation. These results demonstrate that the GPP1 gene plays an important role in the osmoregulation in C. albicans. PMID- 15796988 TI - Direct transformation of a clinical isolate of Candida parapsilosis using a dominant selection marker. AB - Candida parapsilosis is a human pathogenic fungus with increasing importance, particularly in nosocomial infections. For detailed molecular genetic explorations of prototrophic clinical isolates of C. parapsilosis, we developed an efficient transformation system based on a dominant selectable marker. The gene encoding resistance to mycophenolic acid (MPA) was used for selection in yeast transformation. C. parapsilosis cells were transformed with a plasmid vector containing the Candida albicans inosine monophosphate dehydrogenase gene (IMH3) responsible for mycophenolic acid resistance. Transformation was carried out both by electroporation and by the lithium acetate (LiAc) method. The LiAc method resulted in very poor transformation efficiency, while the modified electroporation method yielded a high number of mitotically stable transformants exhibiting unambiguous MPA resistance. Two hundred transformants were analysed for the presence of the C. albicans IMH3(r) gene by polymerase chain reaction. Integration of single or multiple plasmid copies into the genomic DNA of C. parapsilosis was determined by Southern hybridization. To our knowledge, the present study is the first report about a method based on a dominant selectable marker for the transformation of a prototrophic, clinical isolate of C. parapsilosis. The described technique may prove to be an efficient tool for the examination of the biology and virulence of this pathogenic yeast. PMID- 15796989 TI - Chemiluminescence of enterococci isolates from freshwater. AB - All Enterococcus spp., isolated from environmental water samples (n=81), emitted a high chemiluminescence signal in the presence of luminol (10(-2) M). Kinetic studies of chemiluminescence show a close correlation between chemiluminescence and growth curves during the exponential phase, with a maximum chemiluminescence reached just before bacterial growth entered in the stationary phase. On the other hand, genera closely related to Enterococcus such as Streptococcus or Lactococcus produced a very weak chemiluminescent signal. Chemiluminescence of enterococci could therefore offer a rapid test, in aiding the identification of the genus Enterococcus and in the survey of the microbiological quality of water supplies. PMID- 15796990 TI - Identification of mature appressorium-enriched transcripts in Magnaporthe grisea, the rice blast fungus, using suppression subtractive hybridization. AB - We have constructed a fungal subtractive suppressive library enriched in genes expressed during appressorium maturation in Magnaporthe grisea. Sequencing of 250 clones from the subtracted appressorium cDNA library revealed 142 unique genes, represented by 155 non-redundant ESTs (expressed sequence tags). Of these ESTs, 72 have not been previously isolated in M. grisea. RT-PCR analysis of 105 of the genes discovered found transcripts corresponding to 71 of the ESTs only in mature appressoria while transcripts corresponding to a further 34 of the isolated ESTs were expressed both in appressoria and conidia/mycelia. Genes specifically expressed in appressorium identified by SSH included a number that have been previously implicated in appressorium formation or function including GAS1, GAS3, and PTH11. PMID- 15796991 TI - Competition among symbiotic cyanobacterial Nostoc strains forming artificial associations with rice (Oryza sativa). AB - Competition among four symbiotically competent Nostoc strains, colonizing rice roots, was examined using hetR-DGGE (denaturing gradient gel electrophoresis) as strain identification. Although mixed in various combinations, only one strain at a time associated with the rice roots. Nostoc strain 8964:3 was the most competitive and our data suggest that its competitive fitness was dependent on rapid hormogonial spreading as displayed on agar plates. Furthermore, rice roots induced hormogonia in all tested Nostoc strains, but only Nostoc strain 9104 showed positive chemotaxis towards the root. Inhibition of growth of competing cyanobacterial strains was not apparent. PMID- 15796992 TI - Diversity of carbazole-degrading bacteria having the car gene cluster: isolation of a novel gram-positive carbazole-degrading bacterium. AB - Twenty-seven carbazole-utilizing bacterial strains were isolated from environmental samples, and were classified into 14 groups by amplified ribosomal DNA restriction analysis. Southern hybridization analyses showed that 3 and 17 isolates possessed the car gene homologs of Pseudomonas resinovorans CA10 and Sphingomonas sp. strain KA1, respectively. Of the 17 isolates, 2 isolates also have the homolog of the carAa gene of Sphingomonas sp. strain CB3. While the genome of one isolate, a Gram-positive Nocardioides sp. strain IC177, showed no hybridization to any car gene probes, PCR and sequence analyses indicated that strain IC177 had tandemly linked carAa and carC gene homologs whose deduced amino acid sequences showed 51% and 36% identities with those of strain KA1. PMID- 15796993 TI - Interactive optical trapping shows that confinement is a determinant of growth in a mixed yeast culture. AB - Applying a newly developed user-interactive optical trapping system, we controllably surrounded individual cells of one yeast species, Hanseniaspora uvarum, with viable cells of another yeast species, Saccharomyces cerevisiae, thus creating a confinement of the former. Growth of surrounded and non surrounded H. uvarum cells was followed under a microscope by determining their generation time. The average generation time of surrounded H. uvarum cells was 15% higher than that of non-surrounded cells, thereby showing that the confinement imposed by viable S. cerevisiae cells on H. uvarum inhibits growth of the latter. This study is the first to demonstrate that confinement is a determinant of growth in a microbial ecosystem. PMID- 15796994 TI - Evidence for functional laccases in the acidophilic ascomycete Hortaea acidophila and isolation of laccase-specific gene fragments. AB - Hortaea acidophila is a pigmented, yeast-like ascomycete that is able to grow at a pH as low as 0.6. This study presents evidence that H. acidophila possesses at least two functional laccases that seem to be involved in melanin synthesis. This evidence is supported by PCR amplification of laccase-specific gene fragments by using primers derived from conserved copper-binding-regions and by Southern Blot analysis. Due to their low pH optimum the laccases may be of special interest for biotechnological use. PMID- 15796995 TI - Genetic modification of essential fatty acids biosynthesis in Hansenula polymorpha. AB - The Delta(6)-desaturase gene isoform II involved in the formation of gamma linolenic acid (GLA) was identified from Mucor rouxii. To study the possibility of alteration of the synthetic pathway of essential fatty acids in the methylotrophic yeast, Hansenula polymorpha, the cloned gene of M. rouxii under the control of the methanol oxidase (MOX) promoter of H. polymorpha, was used for genetic modification of this yeast. Changes in flux through the n-3 and n-6 pathways in the transgenic yeast were observed. The proportion of GLA varied dramatically depending on the growth temperature and media composition. This can be explained by the effects of either substrate availability or enzymatic activity. In addition to the potential application for manipulating the fatty acid profile, this study provides an attractive model system of H. polymorpha for investigating the deviation of fatty acid metabolism in eukaryotes. PMID- 15796996 TI - Multilocus sequence typing reveals that Bacillus cereus strains isolated from clinical infections have distinct phylogenetic origins. AB - Eight strains of Bacillus cereus isolated from bacteremia and soft tissue infections were assigned to seven sequence types (STs) by multilocus sequence typing (MLST). Two strains from different locations had identical STs. The concatenated sequences of the seven STs were aligned with 65 concatenated sequences from reference STs and a neighbor-joining tree was constructed. Two strains were distantly related to all reference STs. Three strains were recovered in a clade that included Bacillus anthracis, B. cereus and rare Bacillus thuringiensis strains while the other three strains were assigned to two STs that were more closely affiliated to most of the B. thuringiensis STs. We conclude that invasive B. cereus strains do not form a single clone or clonal complex of highly virulent strains. PMID- 15796997 TI - The prevalence of the Staphylococcus aureus tst gene among community- and hospital-acquired strains and isolates from Wegener's Granulomatosis patients. AB - To allow rapid identification of toxic shock syndrome toxin-1 (TSST-1)-producing Staphylococcus aureus strains, a real-time PCR assay for the detection of the tst gene, which encodes TSST-1, was developed. The assay was applied to S. aureus isolates from patients with Wegener's Granulomatosis (WG), as well as isolates that were classified as either community- (CA) or hospital-acquired (HA). No significant difference in the percentage of tst-positive strains was observed between isolates from WG patients and CA isolates (24% and 25%, respectively). In contrast, only 14% of the HA isolates were tst-positive (p<0.05). Investigation of the clonal relationship between tst-positive CA and HA strains could indicated the recent emergence of a virulent S. aureus clone in the community. PMID- 15796998 TI - A critical step for relative quantification of mRNAs is selecting the correct internal controls. PMID- 15796999 TI - Modelling of classification rules on metabolic patterns including machine learning and expert knowledge. AB - Machine learning has a great potential to mine potential markers from high dimensional metabolic data without any a priori knowledge. Exemplarily, we investigated metabolic patterns of three severe metabolic disorders, PAHD, MCADD, and 3-MCCD, on which we constructed classification models for disease screening and diagnosis using a decision tree paradigm and logistic regression analysis (LRA). For the LRA model-building process we assessed the relevance of established diagnostic flags, which have been developed from the biochemical knowledge of newborn metabolism, and compared the models' error rates with those of the decision tree classifier. Both approaches yielded comparable classification accuracy in terms of sensitivity (>95.2%), while the LRA models built on flags showed significantly enhanced specificity. The number of false positive cases did not exceed 0.001%. PMID- 15797000 TI - Algorithms for rapid outbreak detection: a research synthesis. AB - The threat of bioterrorism has stimulated interest in enhancing public health surveillance to detect disease outbreaks more rapidly than is currently possible. To advance research on improving the timeliness of outbreak detection, the Defense Advanced Research Project Agency sponsored the Bio-event Advanced Leading Indicator Recognition Technology (BioALIRT) project beginning in 2001. The purpose of this paper is to provide a synthesis of research on outbreak detection algorithms conducted by academic and industrial partners in the BioALIRT project. We first suggest a practical classification for outbreak detection algorithms that considers the types of information encountered in surveillance analysis. We then present a synthesis of our research according to this classification. The research conducted for this project has examined how to use spatial and other covariate information from disparate sources to improve the timeliness of outbreak detection. Our results suggest that use of spatial and other covariate information can improve outbreak detection performance. We also identified, however, methodological challenges that limited our ability to determine the benefit of using outbreak detection algorithms that operate on large volumes of data. Future research must address challenges such as forecasting expected values in high-dimensional data and generating spatial and multivariate test data sets. PMID- 15797001 TI - Modeling a description logic vocabulary for cancer research. AB - The National Cancer Institute has developed the NCI Thesaurus, a biomedical vocabulary for cancer research, covering terminology across a wide range of cancer research domains. A major design goal of the NCI Thesaurus is to facilitate translational research. We describe: the features of Ontylog, a description logic used to build NCI Thesaurus; our methodology for enhancing the terminology through collaboration between ontologists and domain experts, and for addressing certain real world challenges arising in modeling the Thesaurus; and finally, we describe the conversion of NCI Thesaurus from Ontylog into Web Ontology Language Lite. Ontylog has proven well suited for constructing big biomedical vocabularies. We have capitalized on the Ontylog constructs Kind and Role in the collaboration process described in this paper to facilitate communication between ontologists and domain experts. The artifacts and processes developed by NCI for collaboration may be useful in other biomedical terminology development efforts. PMID- 15797002 TI - A Bayesian network coding scheme for annotating biomedical information presented to genetic counseling clients. AB - We developed a Bayesian network coding scheme for annotating biomedical content in layperson-oriented clinical genetics documents. The coding scheme supports the representation of probabilistic and causal relationships among concepts in this domain, at a high enough level of abstraction to capture commonalities among genetic processes and their relationship to health. We are using the coding scheme to annotate a corpus of genetic counseling patient letters as part of the requirements analysis and knowledge acquisition phase of a natural language generation project. This paper describes the coding scheme and presents an evaluation of intercoder reliability for its tag set. In addition to giving examples of use of the coding scheme for analysis of discourse and linguistic features in this genre, we suggest other uses for it in analysis of layperson oriented text and dialogue in medical communication. PMID- 15797003 TI - Prospective recruitment of patients with congestive heart failure using an ad-hoc binary classifier. AB - This paper addresses a very specific problem of identifying patients diagnosed with a specific condition for potential recruitment in a clinical trial or an epidemiological study. We present a simple machine learning method for identifying patients diagnosed with congestive heart failure and other related conditions by automatically classifying clinical notes dictated at Mayo Clinic. This method relies on an automatic classifier trained on comparable amounts of positive and negative samples of clinical notes previously categorized by human experts. The documents are represented as feature vectors, where features are a mix of demographic information as well as single words and concept mappings to MeSH and HICDA classification systems. We compare two simple and efficient classification algorithms (Naive Bayes and Perceptron) and a baseline term spotting method with respect to their accuracy and recall on positive samples. Depending on the test set, we find that Naive Bayes yields better recall on positive samples (95 vs. 86%) but worse accuracy than Perceptron (57 vs. 65%). Both algorithms perform better than the baseline with recall on positive samples of 71% and accuracy of 54%. PMID- 15797004 TI - Identifying reasoning strategies in medical decision making: a methodological guide. AB - Reasoning strategies are a key component in many medical tasks, including decision making, clinical problem solving, and understanding of medical texts. Identification of reasoning strategies used by clinicians may prove critical to the optimal design of decision support systems. This paper presents a formal method of cognitive-semantic analysis for the identification and characterization of reasoning strategies deployed in medical tasks and demonstrates its use through specific examples. Although semantic analysis was originally developed in the investigation of knowledge structures, it can also be applied to identify the reasoning and decision processes used by physicians and medical trainees in clinical tasks. Assumptions underlying the methods, as well as illustrations of their use in diagnostic explanation tasks, are presented. We discuss semantic analysis in the context of the current interests in developing medical ontologies and argue that a frame-based propositional analytic methodology can provide a systematic way of addressing the construction of such ontologies. Although the application of propositional analysis methods has some limitations, we show how such limitations are being addressed and present some examples of information tools that have been developed to ease, and make more systematic, the process of analysis. PMID- 15797005 TI - Common ground. PMID- 15797006 TI - Parent-offspring conflict. PMID- 15797007 TI - Female-biased gene expression in the malaria mosquito Anopheles gambiae. PMID- 15797008 TI - Integrative biology: sea hares saved by a delicious distraction. AB - When threatened, sea hares secrete ink and opaline. This mixture has now been shown to act on peripheral chemosensory neurons of spiny lobsters, stimulating feeding-related behaviours as a deceptive method of avoiding predation. PMID- 15797009 TI - Legume haemoglobins: symbiotic nitrogen fixation needs bloody nodules. AB - How do plants create an environment in which symbiotic bacteria can reduce enough N2 to provide the plant with sufficient ammonium for growth? Gene silencing has now been used to show that legume haemoglobins are crucial. PMID- 15797010 TI - Neuronal polarity: until GSK-3 do us part. AB - Specification of the axon and dendrites is a critical step in the development of a neuron. Two new studies have shed light on the molecular pathway that controls the establishment of neuronal polarity. PMID- 15797011 TI - Homologous recombination: needing to have my say. AB - Recent studies in budding and fission yeasts have revealed Mei5 and Sae3 as factors necessary for the proper function of the recombinases Dmc1 and Rad51 in DNA repair and meiotic recombination, providing new insights into how strand exchange proteins are directed along specific recombination pathways. PMID- 15797012 TI - Synaptogenesis: a balancing act between excitation and inhibition. AB - Recent studies have implicated a number of membrane-associated proteins, including the signaling pair neuroligin and beta-neurexin, in synapse formation, suggesting that they govern the ratio of inhibitory and excitatory synapses on CNS neurons. These findings, together with data indicating that the genes encoding neuroligin and PSD95 are altered in autism patients, suggest that a molecular understanding of complex neurological diseases is within reach. PMID- 15797013 TI - Visual cortex: two-photon excitement. AB - Current in vivo methods for imaging the visual cortex lack the ability to map response properties at the level of single cells. A new technique using two photon imaging of calcium signals has now overcome this limitation. PMID- 15797014 TI - Plant development: auxin in loops. AB - Concentration gradients of the hormone auxin are associated with various patterning events in plants. Recent work has refined our picture of the complex and dynamic system of auxin transport underlying the formation of these gradients. PMID- 15797015 TI - Eph receptors: two ways to sharpen boundaries. AB - Eph receptors and ephrins can sharpen domains within developing tissues by mediating repulsion at interfaces. An Eph receptor has now been shown also to regulate cell adhesion within tissue subdivisions. PMID- 15797016 TI - Conserved patterns of cell movements during vertebrate gastrulation. AB - Vertebrate embryogenesis entails an exquisitely coordinated combination of cell proliferation, fate specification and movement. After induction of the germ layers, the blastula is transformed by gastrulation movements into a multilayered embryo with head, trunk and tail rudiments. Gastrulation is heralded by formation of a blastopore, an opening in the blastula. The axial side of the blastopore is marked by the organizer, a signaling center that patterns the germ layers and regulates gastrulation movements. During internalization, endoderm and mesoderm cells move via the blastopore beneath the ectoderm. Epiboly movements expand and thin the nascent germ layers. Convergence movements narrow the germ layers from lateral to medial while extension movements elongate them from head to tail. Despite different morphology, parallels emerge with respect to the cellular and genetic mechanisms of gastrulation in different vertebrate groups. Patterns of gastrulation cell movements relative to the blastopore and the organizer are similar from fish to mammals, and conserved molecular pathways mediate gastrulation movements. PMID- 15797017 TI - The Sp1-related transcription factors sp5 and sp5-like act downstream of Wnt/beta catenin signaling in mesoderm and neuroectoderm patterning. AB - BACKGROUND: Wnt/beta-catenin signaling regulates many processes during vertebrate development, including patterning of the mesoderm along the dorso-ventral axis and patterning of the neuroectoderm along the anterior-posterior axis during gastrulation. However, relatively little is known about Wnt target genes mediating these effects. RESULTS: Using zebrafish DNA microarrays, we have identified several new targets of Wnt/beta-catenin signaling, including sp5-like (sp5l, previously called spr2), a zinc-finger transcription factor of the Sp1 family. sp5-like is a direct target of Wnt/beta-catenin signaling and acts together with its paralog sp5 (previously called bts1) downstream of wnt8 in patterning of the mesoderm and neuroectoderm because (1) overexpression of sp5 like, like overexpression of wnt8, posteriorizes the neuroectoderm, (2) sp5-like morpholino-mediated knockdown, like wnt8 knockdown, causes anteriorization of the hindbrain, (3) combined knockdown of sp5 and sp5-like, like loss of wnt8, causes expansion of dorsal mesoderm, (4) sp5-like knockdown reduces the defects in mesoderm and neuroectoderm patterning caused by wnt8 overexpression, and (5) inhibition of sp5-like enhances the effects of hypomorphic loss of wnt8. Importantly, (6) overexpression of sp5-like is able to partially restore normal hindbrain patterning in wnt8 morphants. CONCLUSIONS: sp5-like is a direct target of Wnt/beta-catenin signaling during gastrulation and, together with sp5, acts as a required mediator of the activities of wnt8 in patterning the mesoderm and neuroectoderm. We conclude that sp5 transcription factors mediate the downstream responses to Wnt/beta-catenin signaling in several developmental processes in zebrafish. PMID- 15797018 TI - The noncoding RNA taurine upregulated gene 1 is required for differentiation of the murine retina. AB - BACKGROUND: With the advent of genome-wide analyses, it is becoming evident that a large number of noncoding RNAs (ncRNAs) are expressed in vertebrates. However, of the thousands of ncRNAs identified, the functions of relatively few have been established. RESULTS: In a screen for genes upregulated by taurine in developing retinal cells, we identified a gene that appears to be a ncRNA. Taurine Upregulated Gene 1 (TUG1) is a spliced, polyadenylated RNA that does not encode any open reading frame greater than 82 amino acids in its full-length, 6.7 kilobase (kb) RNA sequence. Analyses of Northern blots and in situ hybridization revealed that TUG1 is expressed in the developing retina and brain, as well as in adult tissues. In the newborn retina, knockdown of TUG1 with RNA interference (RNAi) resulted in malformed or nonexistent outer segments of transfected photoreceptors. Immunofluorescent staining and microarray analyses suggested that this loss of proper photoreceptor differentiation is a result of the disregulation of photoreceptor gene expression. CONCLUSIONS: A function for a newly identified ncRNA, TUG1, has been established. TUG1 is necessary for the proper formation of photoreceptors in the developing rodent retina. PMID- 15797019 TI - erbb3 and erbb2 are essential for schwann cell migration and myelination in zebrafish. AB - BACKGROUND: Myelin is critical for efficient axonal conduction in the vertebrate nervous system. Neuregulin (Nrg) ligands and their ErbB receptors are required for the development of Schwann cells, the glial cells that form myelin in the peripheral nervous system. Previous studies have not determined whether Nrg-ErbB signaling is essential in vivo for Schwann cell fate specification, proliferation, survival, migration, or the onset of myelination. RESULTS: In genetic screens for mutants with disruptions in myelinated nerves, we identified mutations in erbb3 and erbb2, which together encode a heteromeric tyrosine kinase receptor for Neuregulin ligands. Phenotypic analysis shows that both genes are essential for development of Schwann cells. BrdU-incorporation studies and time lapse analysis reveal that Schwann cell proliferation and migration, but not survival, are disrupted in erbb3 mutants. We show that Schwann cells can migrate in the absence of DNA replication. This uncoupling of proliferation and migration indicates that erbb gene function is required independently for these two processes. Pharmacological inhibition of ErbB signaling at different stages reveals a continuing requirement for ErbB function during migration and also provides evidence that ErbB signaling is required after migration for proliferation and the terminal differentiation of myelinating Schwann cells. CONCLUSIONS: These results provide in vivo evidence that Neuregulin-ErbB signaling is essential for directed Schwann cell migration and demonstrate that this pathway is also required for the onset of myelination in postmigratory Schwann cells. PMID- 15797020 TI - Ganglion cells are required for normal progenitor- cell proliferation but not cell-fate determination or patterning in the developing mouse retina. AB - The vertebrate retina develops from an amorphous sheet of dividing retinal progenitor cells (RPCs) through a sequential process that culminates in an exquisitely patterned neural tissue. A current model for retinal development posits that sequential cell-type differentiation is the result of changes in the intrinsic competence state of multipotent RPCs as they advance in time and that the intrinsic changes are influenced by continuous changes in the extracellular environment. Although several studies support the proposition that newly differentiated cells alter the extrinsic state of the developing retina, it is still far from clear what role they play in modifying the extracellular environment and in influencing the properties of RPCs. Here, we specifically ablate retinal ganglion cells (RGCs) as they differentiate, and we determine the impact of RGC absence on retinal development. We find that RGCs are not essential for changing the competence of RPCs, but they are necessary for maintaining sufficient numbers of RPCs by regulating cell proliferation via growth factors. Intrinsic rather than extrinsic factors are likely to play the critical roles in determining retinal cell fate. PMID- 15797021 TI - Symbiotic leghemoglobins are crucial for nitrogen fixation in legume root nodules but not for general plant growth and development. AB - Hemoglobins are ubiquitous in nature and among the best-characterized proteins. Genetics has revealed crucial roles for human hemoglobins, but similar data are lacking for plants. Plants contain symbiotic and nonsymbiotic hemoglobins; the former are thought to be important for symbiotic nitrogen fixation (SNF). In legumes, SNF occurs in specialized organs, called nodules, which contain millions of nitrogen-fixing rhizobia, called bacteroids. The induction of nodule-specific plant genes, including those encoding symbiotic leghemoglobins (Lb), accompanies nodule development. Leghemoglobins accumulate to millimolar concentrations in the cytoplasm of infected plant cells prior to nitrogen fixation and are thought to buffer free oxygen in the nanomolar range, avoiding inactivation of oxygen-labile nitrogenase while maintaining high oxygen flux for respiration. Although widely accepted, this hypothesis has never been tested in planta. Using RNAi, we abolished symbiotic leghemoglobin synthesis in nodules of the model legume Lotus japonicus. This caused an increase in nodule free oxygen, a decrease in the ATP/ADP ratio, loss of bacterial nitrogenase protein, and absence of SNF. However, LbRNAi plants grew normally when fertilized with mineral nitrogen. These data indicate roles for leghemoglobins in oxygen transport and buffering and prove for the first time that plant hemoglobins are crucial for symbiotic nitrogen fixation. PMID- 15797022 TI - EphA4 is required for cell adhesion and rhombomere-boundary formation in the zebrafish. AB - The formation of boundaries between or within tissues is a fundamental aspect of animal development. In the developing vertebrate hindbrain, boundaries separate molecularly and neuroanatomically distinct segments called rhombomeres. Transplantation studies have suggested that rhombomere boundaries form by the local sorting out of cells with different segmental identities. This sorting-out process has been shown to involve repulsive interactions between cells expressing an Eph receptor tyrosine kinase, EphA4, and cells expressing its ephrinB ligands. Although a model for rhombomere-boundary formation based on repulsive Eph-ephrin signaling is well established in the literature, the predictions of this model have not been tested in loss-of-function experiments. Here, we eliminate EphA4 and ephrinB2a proteins in zebrafish with antisense morpholinos (MO) and find that rhombomere boundaries are disrupted in EphA4MO embryos, consistent with a requirement for Eph-ephrin signaling in boundary formation. However, in mosaic embryos, we observe that EphA4MO cells and EphA4-expressing cells sort from one another, an observation that is not predicted by the Eph-ephrin repulsion model but instead suggests that EphA4 promotes cell adhesion within the rhombomeres in which it is expressed. Differential cell adhesion is known to be an effective mechanism for cell sorting. We therefore propose that the well-known EphA4 dependent repulsion between rhombomeres operates in parallel with the EphA4 dependent adhesion within rhombomeres described here to drive the cell sorting that underlies rhombomere-boundary formation. PMID- 15797023 TI - Monkeys pay per view: adaptive valuation of social images by rhesus macaques. AB - Individuals value information that improves decision making. When social interactions complicate the decision process, acquiring information about others should be particularly valuable. In primate societies, kinship, dominance, and reproductive status regulate social interactions and should therefore systematically influence the value of social information, but this has never been demonstrated. Here, we show that monkeys differentially value the opportunity to acquire visual information about particular classes of social images. Male rhesus macaques sacrificed fluid for the opportunity to view female perinea and the faces of high-status monkeys but required fluid overpayment to view the faces of low-status monkeys. Social value was highly consistent across subjects, independent of particular images displayed, and only partially predictive of how long subjects chose to view each image. These data demonstrate that visual orienting decisions reflect the specific social content of visual information and provide the first experimental evidence that monkeys spontaneously discriminate images of others based on social status. PMID- 15797024 TI - Sea hares use novel antipredatory chemical defenses. AB - Numerous studies have demonstrated that chemical defenses protect prey from predation and have often assumed that these defenses function by repelling predators. Surprisingly, few have investigated the mechanisms whereby predators are affected by these defenses. Here, we examine mechanisms of chemical defense of sea hares (Aplysia californica), which, when attacked by spiny lobsters (Panulirus interruptus), release defensive secretions from ink and opaline glands. We show that ink-opaline facilitates the escape of sea hares by acting through a combination of novel and conventional mechanisms. Ink-opaline contains millimolar quantities of amino acids that stimulate chemoreceptor neurons in the spiny lobster's nervous system. Ink stimulates appetitive and ingestive behavior, opaline can elicit appetitive behavior but can also inhibit ingestion and evoke escape responses, and both stimulate grooming. These results suggest that these secretions function by "phagomimicry," in which ink-opaline stimulates the feeding pathway to deceive spiny lobsters into attending to a false food stimulus, and by sensory disruption, in which the sticky and potent secretions cause high-amplitude, long-lasting chemo-mechanosensory stimulation. In addition, opaline contains a chemical deterrent that opposes appetitive effects. Thus, chemical defenses may act in more complex manners than palatability assays of prey chemistry may suggest. PMID- 15797025 TI - Conversion of functional specificity in Qb-SNARE VTI1 homologues of Arabidopsis. AB - In higher multicellular eukaryotes, highly specialized membrane structures or membrane trafficking events are required for supporting various physiological functions. SNAREs (soluble NSF attachment protein receptors) play an important role in specific membrane fusions. These protein receptors are assigned to subgroubs (Qa-, Qb-, Qc-, and R-SNARE) according to their specific SNARE structural motif. A specific set of Qa-, Qb-, and Qc-SNAREs, located on the target membrane, interact with R-SNARE on the vesicle to form a tight complex, leading to membrane fusion. The zig-1 mutant of Arabidopsis lacking Qb-SNARE VTI11 shows little shoot gravitropism and abnormal stem morphology. VTI11 and its homolog VTI12 exhibit partially overlapping but distinct intracellular localization and have different biological functions in plants. Little is known about how SNAREs are targeted to specific organelles, even though their functions and specific localization are closely linked. Here, we report that a novel mutation in VTI12 (zip1) was found as a dominant suppressor of zig-1. The zip1 mutation gave VTI12 the ability to function as VTI11 by changing both the specificity of SNARE complex formation and its intracellular localization. One amino acid substitution drastically altered VTI12, allowing it to suppress abnormalities of higher order physiological functions such as gravitropism and morphology. The zip1 mutation may be an indication of the flexibility in plant cell function afforded by gene duplication, particularly among the VTI11 genes and their recently diverged orthologs. PMID- 15797026 TI - Cell-nonautonomous induction of ovarian and uterine serous cystadenomas in mice lacking a functional Brca1 in ovarian granulosa cells. AB - Women with germline mutations in BRCA1 have a 40% risk of developing ovarian cancer by age 70 and are also predisposed to cancers of the fallopian tubes. Given that ovulatory activity is a strong risk factor for sporadic ovarian cancer, we hypothesized that reduced BRCA1 expression might predispose to gynecological cancers indirectly, by influencing ovarian granulosa cells. These cells secrete sex steroids that control the ovulatory cycle and influence the growth of ovarian epithelial tumors. Granulosa cells also secrete mullerian inhibiting substance (MIS), a hormone that inhibits both the formation of female reproductive organs in male embryos and the proliferation of ovarian epithelial tumor cells. We tested this hypothesis by using the Cre-lox system to inactivate the Brca1 gene in mouse ovarian granulosa cells. A truncated form of the Fsh receptor promoter served as the Cre driver. Here, we show that indeed, inactivation of the Brca1 gene in granulosa cells led to the development of cystic tumors in the ovaries and uterine horns. These tumors carried normal Brca1 alleles, supporting the view that Brca1 may influence tumor development indirectly, possibly through an effector secreted by granulosa cells. PMID- 15797027 TI - p84, a new Gbetagamma-activated regulatory subunit of the type IB phosphoinositide 3-kinase p110gamma. AB - A variety of genetic and inhibitor studies have shown that phosphoinositide 3 kinase gamma (PI3Kgamma) plays an essential role in a number of physiological responses, including neutrophil chemotaxis, mast cell degranulation, and cardiac function []. PI3Kgamma is currently thought to be composed of a p110gamma catalytic subunit and a single regulatory subunit, p101. The binding of p110gamma to p101 dramatically increases the activation of the complex by Gbetagamma subunits and, hence, is thought to be critical for the coupling of PI3Kgamma to G protein coupled receptors []. Here, we characterize a new regulatory subunit for PI3Kgamma. p84 is present in human, mouse, chicken, frog, and fugu genomes and is located beside the p101 locus. It is broadly expressed in cells of the murine immune system. Both recombinant and endogenous p84 bind p110gamma specifically and with high affinity. Binding of p84 to p110gamma substantially increases the ability of Gbetagamma to stimulate phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P(3)) production both in vitro and in vivo. However, the p84/p110gamma heterodimer is approximately 4-fold less sensitive to Gbetagammas than p101/p110gamma. Endogenous murine p84 expression is substantially reduced in the absence of p110gamma expression. We conclude that p110gamma has two potential regulatory subunits in vivo, p84 and p101. PMID- 15797028 TI - Response to visual threat following damage to the pulvinar. AB - We present a unique case demonstrating contributions of the pulvinar in response to visual threat. Substantial evidence demonstrates that the amygdala contributes to the emotion of fear and the response to threat. Traditionally, two routes to amygdala activation have been distinguished: a "slow cortical" route through visual and association cortex and a "fast subcortical" route through the thalamus. The pulvinar nucleus of the thalamus is well connected to the amygdala, suggesting that pulvinar damage might interfere with amygdala activation and response to threat. We tested this possibility in patient SM, who suffered complete loss of the left pulvinar. We measured interference from threatening images on goal-directed behavior. In SM's ipsilesional field, threatening images slowed responses more than pleasant images did. This interference decreased rapidly over time. In contrast, in SM's contralesional field, interference from threatening images was initially absent and then increased rather than decreased over time. Processing through the pulvinar therefore plays a significant role in generating response to visual threat. We suggest that, with disruption of the subcortical route to the amygdala, briefly presented images were not fully processed for threat. The reemergence of interference over time may reflect contributions of a slower route. PMID- 15797029 TI - Contextual modulation outside of awareness. AB - Contextual effects are ubiquitous in vision and reveal fundamental principles of sensory coding. Here, we demonstrate that an oriented surround grating can affect the perceived orientation of a central test grating even when backward masking of the surround prevents its orientation from being consciously perceived. The effect survives introduction of a gap between test and surround of over a degree even under masking, suggesting either that contextual information can effectively propagate across early visual cortex in the absence of awareness of the signaled context or that it can proceed undetected to higher processing levels at which such horizontal propagation may not be necessary. The effect under masking also shows partial interocular transfer, demonstrating processing of orientation by binocular neurons in visual cortex in the absence of conscious orientation perception. This pattern of results is consistent with the suggestion that simultaneous orientation contrast is mediated at multiple levels of the visual processing hierarchy, and it supports the view that propagation of signals to and, possibly, back from higher visual areas is necessary for conscious perception. PMID- 15797030 TI - Transitions in ancient inland freshwater resource management in Sri Lanka affect biota and human populations in and around coastal lagoons. AB - The increasing anthropogenic pressure on natural environments results in impacts that affect tropical forest areas and their biodiversity. Adverse impacts on terrestrial and oceanic environments often compound in the intertidal area, where mangrove forest ecosystems thrive. In tropical coastal areas of many developing countries where people depend on wood and other mangrove forest products and services, forest degradation leads to socioeconomic problems. At the same time, increasing freshwater needs in these areas are expected to cause additional problems. On the basis of remote sensing and ground truthing complemented by colonial archival material from the Dutch East India Company (1602-1800), we report that changes to the historic system of inland freshwater management have increased dramatically in recent times. Hydrological changes, such as interbasin transfers, have resulted in a qualitative ecological and socioeconomic degradation in three coastal lagoons in southern Sri Lanka. Variations in river hydrology have caused changes in the areas suitable as mangrove habitat and, thus, have resulted in an altered distribution. However, increases in mangrove area can mask the degradation of the site in terms of floristic composition, significance of the species, and biodiversity (this effect is termed "cryptic ecological degradation"). It is important that such changes be carefully monitored to ensure biological and socioeconomic sustainability. PMID- 15797032 TI - Cardiac assistance from skeletal muscle: should we be downhearted? PMID- 15797031 TI - The RNA binding protein TLS is translocated to dendritic spines by mGluR5 activation and regulates spine morphology. AB - Neuronal dendrites, together with dendritic spines, exhibit enormously diverse structure. Selective targeting and local translation of mRNAs in dendritic spines have been implicated in synapse remodeling or synaptic plasticity. The mechanism of mRNA transport to the postsynaptic site is a fundamental question in local dendritic translation. TLS (translocated in liposarcoma), previously identified as a component of hnRNP complexes, unexpectedly showed somatodendritic localization in mature hippocampal pyramidal neurons. In the present study, TLS was translocated to dendrites and was recruited to dendrites not only via microtubules but also via actin filaments. In mature hippocampal pyramidal neurons, TLS accumulated in the spines at excitatory postsynapses upon mGluR5 activation, which was accompanied by an increased RNA content in dendrites. Consistent with the in vitro studies, TLS-null hippocampal pyramidal neurons exhibited abnormal spine morphology and lower spine density. Our results indicate that TLS participates in mRNA sorting to the dendritic spines induced by mGluR5 activation and regulates spine morphology to stabilize the synaptic structure. PMID- 15797033 TI - Quantitative methods for tracking cognitive change 3 years after coronary artery bypass surgery. AB - The analysis and interpretation of change in cognitive function test scores after coronary artery bypass grafting (CABG) present considerable statistical challenges. Application of hierarchical linear statistical models can estimate the effects of a surgical intervention on the time course of multiple biomarkers. We use an "analyze then summarize" approach whereby we estimate the intervention effects separately for each cognitive test and then pool them, taking appropriate account of their statistical correlations. The model accounts for dropouts at follow-up, the chance of which may be related to past cognitive score, by implicitly imputing the missing data from individuals' past scores and group patterns. We apply this approach to a study of the effects of CABG on the time course of cognitive function as measured by 16 separate neuropsychological test scores, clustered into 8 cognitive domains. The study includes measurements on 140 CABG patients and 92 nonsurgical controls at baseline, and at 3, 12, and 36 months. Our "analyze then summarize" method allows us to identify differences between the treatment groups in individual tests as well as in aggregate measures. It takes into account the correlation structure of the data and thereby produces more precise results than summarizing before analyzing. The methods used have application to a wide range of intervention studies in which multiple biomarkers are followed over time to quantify health effects. Software to implement the methods in the R statistical package is available from the authors at http://www.biostat.jhsph.edu/sbarry/software/ATSrcode.pdf. PMID- 15797034 TI - Surgical treatment of esophageal high-grade dysplasia. AB - BACKGROUND: Barrett's esophagus, high-grade dysplasia (HGD), and invasive cancer are steps in the progression of esophageal adenocarcinoma. While surgery is recommended for resectable invasive adenocarcinoma, a number of treatment modalities are advocated for HGD. The purpose of this study is to determine the outcomes after surgery for HGD. METHODS: We identified cases of HGD based on endoscopic biopsy in a single institution's databases from 1980 through 2001. Records were reviewed for patient characteristics, treatments, staging, and outcomes. RESULTS: In a 22-year period, 869 cases of esophageal adenocarcinoma and 1,614 cases of Barrett's esophagus were diagnosed. Of these, 115 had HGD without pretreatment evidence of invasion. Forty-nine patients with HGD underwent resection (mean age, 59 years) as initial treatment. Forty-seven had endoscopic treatment (mean age, 70 years) by photodynamic therapy or endoscopic mucosal resection. Seven of the endoscopically treated patients failed, with three undergoing surgery and four observation. Nineteen patients were initially observed, with six eventually having surgery. For the 49 initially treated surgically, one (2%) operative mortality occurred. Invasive adenocarcinoma was present in 18 (37%). The five-year survival was 83% for all resected HGD patients (91% for those without invasion, 68% with invasion). Three of the eight deaths in those with invasion were from recurrent adenocarcinoma. CONCLUSIONS: Surgical resection of esophageal HGD can be performed with low mortality and allows long term survival. A significant percentage with an initial diagnosis of HGD will have invasive disease at resection. Surgery is the optimal treatment for HGD unless contraindicated by severe comorbidities. PMID- 15797035 TI - A retrospective analysis of locally advanced esophageal cancer patients treated with neoadjuvant chemoradiation therapy followed by surgery or surgery alone. AB - BACKGROUND: We conducted an institutional review of patients with locally advanced esophageal cancer who had complete pretreatment and surgical staging to identify variables predictive of outcome. METHODS: From 1993 through 2002, 286 patients presented for surgical therapy of esophageal cancer. Of these, 176 patients met criteria for review including pretreatment endoscopic ultrasound stages IIA through IVA and a transthoracic surgical approach with "two-field" lymph node dissection. This cohort was primarily male (84.7%, n = 149) with adenocarcinoma (88.6%, n = 156), and 101 patients (57.3%) demonstrated endoscopic ultrasound stage III or IVA. RESULTS: Eighty-five (48.3%) patients presented to surgery after receiving neoadjuvant chemoradiation therapy, and 91 (51.7%) underwent surgery alone. Both groups were well matched with respect to comorbidities and pretreatment stage. Patients receiving neoadjuvant chemoradiation demonstrated a nonsignificant trend toward increased operative mortality and nonfatal morbidity. The overall median survival was 16.8 months, and there was no survival difference comparing patients treated with neoadjuvant chemoradiation followed by surgery or surgery alone (p = 0.82). The subset of 25 patients (29.4%) demonstrating a complete pathologic response after neoadjuvant chemoradiation therapy however had superior survival (median survival = 57.6 months, p < 0.01) as compared with neoadjuvant chemoradiation patients demonstrating partial downstaging (n = 36, 42.3%), no downstaging (n = 24, 28.2%), and surgery alone patients. Multivariate analysis identified a complete pathologic response, endoscopic ultrasound stage, and number of pathologically positive lymph nodes as independent predictors of survival. CONCLUSIONS: These data support the use of neoadjuvant chemoradiation for locally advanced esophageal cancer as the subset of patients who demonstrate a complete pathologic response experienced significantly better survival. PMID- 15797036 TI - Esophageal leiomyoma: a 40-year experience. AB - BACKGROUND: Esophageal leiomyomas, although infrequent, are the most common benign intramural tumors of the esophagus. They represent 10% of all gastrointestinal leiomyomas and frequently cause symptoms, necessitating resection. METHODS: The Massachusetts General Hospital Pathologic Database was reviewed over a 40-year period for patients who underwent surgical resection of esophageal leiomyomas. Data analyzed included demographic information, presenting symptoms, tumor location, tumor characteristics and histology, diagnostic procedures, and treatment modalities/outcomes. Fifty-three patients were identified; 31 patients were symptomatic from their leiomyomas. RESULTS: Symptomatic patients presented at a mean age of 44 years old and exhibited a twofold male predominance. Mean tumor diameter among symptomatic patients was 5.3 cm, as compared to 1.5 cm in asymptomatic patients (p < 0.0001). Thirty of the symptomatic patients had solitary leiomyomas, and 1 patient had five separate leiomyomas. Eighty-four percent of the lesions in symptomatic patients occurred in the lower two-thirds of the esophagus, with epigastric discomfort being the most common presenting symptom. Among patients operated on solely for leiomyoma, 97% were enucleated without an esophageal resection. None of the leiomyomas showed malignant transformation or recurrence. All symptomatic patients had relief of symptoms, with no perioperative morbidity or mortality. CONCLUSIONS: In a large pathologic series, over half of all patients with esophageal leiomyomas were symptomatic. Larger tumors were significantly more likely to be symptomatic. Local enucleation by a variety of surgical approaches was accomplished in most patients. All symptomatic patients had relief of symptoms, with no perioperative morbidity or mortality. There was no observed tendency for malignant transformation or recurrence. PMID- 15797037 TI - Effect of lower esophageal sphincter distension and acidification on esophageal pressure and electromyographic activity: the identification of the "sphinctero esophageal excitatory reflex". AB - BACKGROUND: The mechanism of esophageal acid clearance through augmenting esophageal peristalsis in response to gastroesophageal reflux (GER) is not exactly known. We investigated the hypothesis that lower esophageal sphincter (LES) dilatation rather than the refluxed acid affected reflex increase in the esophageal peristaltic activity aiming at clearing the esophagus of the refluxed acid. METHODS: The esophageal pressure and electromyographic (EMG) activity response to esophageal sphincter balloon distension in increments of 2 mL of saline was recorded in 17 healthy volunteers (10 men, 7 women, mean age 43.6 +/- 11.2 years). The test was repeated in 10 of 17 volunteers while the esophageal sphincter was being anesthetized. The response of the esophageal pressure and electromyographic activity to lower esophageal acidification was also tested. RESULTS: Lower esophageal sphincter balloon distension with 2 mL of saline produced esophageal pressure increase to a mean of 34.2 +/- 5.3 cm H2O (p < 0.001). Increase of the balloon distending volume produced results similar to the 2-mL distension (p > 0.05). The esophageal electrical activity increased on esophageal balloon distension; the increase was similar with distensions of 2 mL up to 10 mL. There was no esophageal pressure or electrical activity response to distension of the anesthetized lower esophageal sphincter or to lower esophageal sphincter acidification. CONCLUSIONS: During gastroesophageal reflux episodes, the lower esophageal sphincter dilatation and not acidification appears to initiate increased esophageal peristalsis, which clears the esophagus of the refluxed acid. The increased esophageal peristalsis on lower esophageal sphincter dilatation is suggested to be reflex in nature and is mediated through the "sphincteroesophageal excitatory reflex." This reflex may be of diagnostic significance in esophageal motility disorders; however, this point needs further studies. PMID- 15797039 TI - Pulmonary carcinoid tumors with Cushing's syndrome: an aggressive variant or not? AB - BACKGROUND: Adrenocorticotropic hormone (ACTH)-secreting pulmonary carcinoid is considered an aggressive variant of carcinoid tumors. Current knowledge is based upon a limited number of reports with few patients. METHODS: All patients with Cushing's syndrome (CS) resulting from pulmonary carcinoid (PC) who underwent pulmonary resection at our institution from November 1966 through April 1998 were reviewed. RESULTS: The group studied consisted of 10 males and 13 females. The median age was 39 years (range: 14-71). Pulmonary symptoms were present in 4 patients. Chest radiographs identified an abnormality in 13 patients (57%) and chest computerized tomography (CT) identified an abnormality in all 20 patients examined. Before pulmonary resection, hypophysectomy and bilateral adrenalectomy were performed in 7 patients (30%) each. Median time interval from presentation to pulmonary resection was 17 months (range: 1-228). Lobectomy was performed in 16 patients, segmentectomy was performed in 4 patients, and bilobectomy, pneumonectomy, and wedge excision was performed in 1 patient each. There were no operative deaths. Typical carcinoid was identified in 21 patients (91%) and atypical carcinoid was identified in 2 patients (9%). The median tumor diameter was 1.3 cm (range: 0.3-10). Nineteen patients (83%) underwent mediastinal lymphadenectomy and lymph node metastasis was found in 6 patients (32%) (N1 in 4 patients, N2 in 2 patients). The median follow-up was 78 months (range: 1-432). CS resolved in all of the patients. CS with PC recurred in 4 patients and CS alone recurred in 1 patient. Two patients underwent curative re-resection. Two patients, one with disseminated PC, died at last follow-up. CONCLUSIONS: Despite long delays in surgical therapy, pulmonary resection for ACTH-secreting PCs results in a favorable prognosis. Anatomic resection with complete mediastinal lymphadenectomy decreases local recurrence. Although rare these tumors do not seem to be as aggressive a variant of typical carcinoid tumors as previously reported. PMID- 15797040 TI - Survival after pathological stage IA nonsmall cell lung cancer: tumor size matters. AB - BACKGROUND: This study evaluates prognostic factors for survival in completely resected pathological stage IA nonsmall cell lung cancer with special emphasis on tumor size and assesses tumor recurrence rate by actual and actuarial analysis. METHODS: From January 1989 to December 2001, 130 consecutive resections for pathological stage IA nonsmall cell lung cancer were performed. Pathological tumor size was categorized into 0 to 20 mm and 21 to 30 mm. Each patient was scaled according to the Charlson Comorbidity Index. The Kaplan-Meier method was used for estimation of actuarial recurrence rate and the cumulative incidence method was used to estimate the actual recurrence rate. Risk factors for overall and disease free survival were determined by univariate and multivariate Cox regression analysis. RESULTS: Overall 5-year survival for patients with tumors 0 to 20 mm and 21 to 30 mm was 69% and 51%, respectively (p = 0.038). Disease-free survival at 5 years was 68% and 48%, respectively (p = 0.015). Only 27 patients had a recurrence and 69 patients died during follow-up. The actual 10-year recurrence rate was lower than the actuarial recurrence rate (23% vs 29%). Larger tumor size (relative risk 1.6; 95% confidence interval 1.0 to 2.7), Charlson Comorbidity Index score greater than or equal to 3 (relative risk 3.7; 95% confidence interval 1.7 to 8.0), and pneumonectomy (relative risk 2.1; 95% confidence interval 1.1 to 4.2) independently predicted adverse outcome. CONCLUSIONS: Tumor size affects survival in resected stage IA nonsmall cell lung cancer. Current definition of stage IA disease should be substaged into two separate stages. In patients with early-stage lung cancer and relatively good prognosis actual recurrence rate is more realistic than the actuarial recurrence rate. PMID- 15797041 TI - Impact of large tumor size on survival after resection of pathologically node negative (pN0) non-small cell lung cancer. AB - BACKGROUND: The current TNM staging system first adopted the tumor size of 3 cm for subdivision of stage I and II disease. The aim of the present study was to evaluate the impact of tumor size on survival in patients with pathologically node negative (pN0) non-small cell lung cancer after complete resection. METHODS: We retrospectively reviewed the records of 603 patients with pN0 non-small cell lung cancer patients (403 men and 200 women) who underwent a complete resection in five national chest hospitals between 1992 and 1996, with follow-up duration of more than 5 years, and analyzed tumor size and survival. Survival rate was estimated by the Kaplan-Meier method, and differences were compared by log-rank test. For the multivariate analysis, the Cox proportional hazard model was used to identify variables that significantly affected survival. RESULTS: There were 355 adenocarcinomas, 208 squamous cell carcinomas, and 40 large cell carcinomas completely resected. No significant prognostic differences were seen among three groups with smaller-sized tumors (< or =2 cm [n = 171], 2.1 to 3 cm [n = 202], and 3.1 to 5 cm [n = 170]); however, patients with a tumor size greater than 5 cm (n = 60) showed a significantly worse prognosis. The 5-year survival rates were 79.6%, 72.7%, 68.1%, and 46.6%, respectively, in these four groups. Multivariate analysis showed the tumor size to be an independent prognostic predictor in patients with pN0 tumors. CONCLUSIONS: We found that a tumor size of greater than 5 cm was an independent prognostic predictor in pN0 disease; therefore, upgrading the T factor of tumor diameter to greater than 5 cm may be necessary in the next reversion of the TNM staging system. PMID- 15797042 TI - Bronchovascular versus bronchial sleeve resection for central lung tumors. AB - BACKGROUND: Pneumonectomy has traditionally been the treatment of choice for central lung tumors. Bronchial sleeve resections are increasingly considered as a reasonable alternative. For tumor involvement of both central airways and pulmonary artery, bronchovascular sleeve resections are possible, but considered to be technically demanding and associated with a higher perioperative risk. In addition, their role as adequate oncologic treatment for lung cancer is unclear. We have compared the early and long-term results of bronchovascular sleeve resection with those of bronchial sleeve resection and pneumonectomy. METHODS: We retrospectively analyzed all patients who underwent bronchial sleeve resection (group I, n = 104), bronchovascular sleeve resection (group II, n = 67), and pneumonectomy (group III, n = 63) for central lung cancer in our institution. RESULTS: The groups were comparable regarding demographics and tumor, node, and metastasis (TNM) stage. Early mortality was 1.9% in group I, 1.5% in group II, and 6.3% in group III (p = 0.19). The rate of bronchial complications was 0.96% in group I, 0% in group II, and 7.9% in group III (p = 0.006). Five-year survival was 46.1% in group I, 42.9% in group II, and 30.4% in group III (p = 0.16). Freedom from local recurrence of disease (5 years) was 83.8% in group I, 84.2% in group II, and 88.7% in group III (p = 0.56). CONCLUSIONS: Bronchovascular sleeve resections are as safe as bronchial sleeve resections for the treatment of central lung cancer. Both procedures have comparable early and long-term results, which are similar to those of pneumonectomy. It appears reasonable to apply bronchovascular sleeve resections more liberally. PMID- 15797043 TI - Local control of disease related to lymph node involvement in non-small cell lung cancer after sleeve lobectomy compared with pneumonectomy. AB - BACKGROUND: Increasing evidence has suggested that sleeve lobectomy might be a viable alternative procedure for pneumonectomy in non-small cell lung cancer (NSCLC), including patients with adequate pulmonary reserve. This study was designed to compare the outcomes of the two procedures and to determine adequate surgical indications for each procedure. METHODS: From January of 1989 to December of 1998, sleeve lobectomy was performed in 49 patients, and 200 patients underwent pneumonectomy for NSCLC. By reviewing the computed tomographic scans, bronchoscopic findings, and operative reports, we selected 49 patients on whom sleeve resection could have been performed. The clinical outcomes of the sleeve lobectomies (SL) and pneumonectomies (PN) were analyzed, particularly in relation to nodal status and recurrence patterns. RESULTS: Operative mortality was 6.1% (3 of 49 patients) in the SL group and 4.1% (2 of 49 patients) in the PN group. Mean follow-up period was 51 months (range, 5 to 149). The overall 5-year survival rate was not substantially different between the two groups (SL: 53.7% vs PN: 59.5%, p = 0.510). Recurrence occurred in 57% (26 of 46 patients) of the SL group and in 30% (14 of 47 patients) of the PN group. The 5-year freedom from recurrence rates were better in the PN group (SL: 45.7% vs PN: 67.9%, p = 0.017). Locoregional recurrences occurred in 32.6% (15 of 46 patients) of the SL group and in 8.5% (4 of 47 patients) of the PN group. In multivariate analysis, performing sleeve resection in patients with a positive N1 lymph node was a significant risk factor for developing locoregional recurrence (p = 0.007). CONCLUSIONS: Although the overall survival rates were similar, sleeve resection resulted in higher locoregional recurrence, particularly in patients with positive N1 lymph nodes. This finding suggests that sleeve resection should be performed in selected patients, such as those without lymph node metastasis. PMID- 15797044 TI - Nocturnal hypoxemia after lobectomy for lung cancer. AB - BACKGROUND: Although nocturnal episodic hypoxemia after major abdominal surgery has been reported, the condition of nocturnal oxygen saturation after lung surgery is largely unknown. We evaluated nocturnal oxygen saturation during the perioperative period after lobectomy for lung cancer. This study also compared the postoperative course of nocturnal oxygen saturation after standard lobectomy with posterolateral thoracotomy and lobectomy with video-assisted thoracic surgery. METHODS: Twenty-one consecutive patients who had undergone lobectomy for lung cancer by either the posterolateral thoracotomy approach (n = 11) or the video-assisted thoracic surgery approach (n = 10) were studied. Fifteen consecutive patients who had undergone gastrectomy for gastric cancer were also studied. Overnight oxygen saturation was measured on the third and 14th postoperative days. RESULTS: The frequency of hypoxemia in the lobectomy group was higher than that in the gastrectomy group (p = 0.043). The frequency of hypoxemia on the 14th postoperative day (p = 0.009) and the severity of hypoxemia on the third and 14th postoperative days (p = 0.041, 0.046) for the video assisted thoracic surgery approach were lower than those for the posterolateral thoracotomy approach. In terms of mean arterial oxygen saturation, heart rate, forced vital capacity, and forced expiratory volume in 1 second, there were no statistically significant differences between the video-assisted thoracic surgery group and the posterolateral thoracotomy group. CONCLUSIONS: Video-assisted thoracic surgery lobectomy was superior in terms of early postoperative nocturnal oxygen saturation. We conclude that the video-assisted thoracic surgery approach is more beneficial than the posterolateral thoracotomy approach for high-risk patients. PMID- 15797045 TI - Characterization and importance of air leak after lobectomy. AB - BACKGROUND: Air leak after pulmonary resection is a common occurrence that is incompletely characterized. Our objectives were to determine prevalence of air leak and identify its risk factors, characterize its duration and discover its correlates, and evaluate its clinical importance. METHODS: Air leak was studied in 319 patients undergoing isolated anatomic lobectomy between January 1998 and July 2001. Risk factors for air leak were identified by logistic regression of patient characteristics, indications for lobectomy, lobe resected, and fissure management. Factors associated with air leak duration were sought by time-related analysis. Association of complications with air leak was evaluated by propensity matched pairs analysis. RESULTS: Prevalence: Air leak prevalence was 58% (186 patients). It occurred less frequently after left lower lobectomy (p < 0.0001) and later in the series (p = 0.008). It was surgeon dependent (p = 0.007) but not associated with forced expiratory volume in 1 second. DURATION: The 10th, 50th, and 90th percentiles of air leak duration were 1.6, 3, and 7 days, respectively. No factors, including fissure management, were reliably associated with air leak duration. IMPORTANCE: Air leak was associated with more complications (30% vs 18%, p = 0.07) and protracted hospital course (p = 0.02). CONCLUSIONS: Postoperative air leak is a common occurrence after lobectomy, but fortunately it is self-limiting in most patients. Air leak is independently associated with longer hospital stay and other postoperative complications. Surgical technique is important and may be the only modifiable factor. PMID- 15797046 TI - The surgical management of superior sulcus tumors: a retrospective review with long-term follow-up. AB - BACKGROUND: We reviewed our experience with multimodality therapy for superior sulcus tumors to identify aspects of treatment that impact survival. METHODS: We retrospectively analyzed the records of 39 consecutive patients who underwent surgical resection in a single institution between 1993 and 2000. RESULTS: Median age at presentation was 59 years (range, 40 to 77). Twenty-five patients (64%) were men. At presentation, 36 patients (92%) had clinical T3 tumors and 3 (8%) had clinical T4 tumors. Mediastinoscopy was negative in all patients. Thirty-one patients (79%) received preoperative radiotherapy (median dose, 4500 cGy). Chemotherapy was administered concurrently with radiotherapy in 27 patients (69%). Complete surgical resection was performed in 34 patients (87%). There were 2 (5%) postoperative deaths. Of the 31 patients who received preoperative therapy, 14 (45%) had their tumors downstaged and 9 (29%) demonstrated a complete pathologic response. Median follow-up (100%) was 69 months. Overall 5-year survival was 47.9%. Five-year survival was 52.1% in patients with negative resection margins (p = 0.005), and 60.6% in patients who demonstrated a response to induction chemoradiation therapy (p = 0.008). Independently, margin status and response to induction therapy are predictors of overall survival (p = 0.01 and p = 0.02, respectively). Multivariable analysis identified margin status as the only factor significantly associated with overall survival. Negative margins strongly correlated with the response to preoperative therapy (p = 0.004). Disease-free survival correlated well with the response to induction therapy (p = 0.03). The chemotherapy regimen, T status, operative procedure, and complete pathologic response did not correlate with survival. CONCLUSIONS: The use of chemoradiation induction therapy may downstage tumors, enhance the ability to obtain a complete surgical resection, and prolong survival. PMID- 15797047 TI - Can aberrant promoter hypermethylation of CpG islands predict the clinical outcome of non-small cell lung cancer after curative resection? AB - BACKGROUND: Aberrant methylation of CpG islands acquired in tumor cells in promoter regions is one cause for the loss of gene function. We examined whether aberrant DNA hypermethylation could be used to predict the clinical outcomes of patients with primary nonsmall cell lung cancer (NSCLC) after curative resection. METHODS: We tested 61 patients with NSCLC using methylation-specific polymerase chain reaction (MSP) and searched for promoter hypermethylation of the genes p16INK4a, retinoic acid receptor beta-promoter (RARbetaP2), death-associated protein kinase (DAPK), and O6-methylguanine-DNA-methyltransferase (MGMT). The clinical data, the presence of DNA hypermethylation, and clinical outcomes were analyzed. RESULTS: Hypermethylation in the tumor samples was detected in 67% (41 of 61) for p16(INK4a), 49% (30 of 61) for RARbetaP2, 30% (18 of 61) for DAPK, and 62% (38 of 61) for MGMT. Thirty patients (49%) developed recurrence within 33 months; 16 in the remaining lung, 10 in other organs, and 4 in both. We found no correlation between the specific DNA hypermethylation and any of the clinicopathological characteristics of the patients. DNA hypermethylation was not associated with a different survival or recurrence rate. However, the aberrant hypermethylation of RARbetaP2 seemed to be related to the location of cancer recurrence. Although advanced T stage and preoperative chemotherapy were statistically significant in univariate analysis, unmethylation of DAPK (p = 0.030) and hypermethylation of RARbetaP2 (p = 0.014), as well as advanced T stage (p = 0.075) and preoperative chemotherapy (p = 0.025), were significant risk factors in multivariate analysis for early recurrence in the remaining lung. CONCLUSIONS: The P2 hypermethylation of the RARbeta gene and unmethylation of DAPK seem to be important factors in predicting early cancer recurrence in the remaining lung and could be used as a prognostic marker in NSCLC. However, the clinical implications of this finding need further investigation. PMID- 15797049 TI - Vacuum-assisted closure for the treatment of complex chest wounds. AB - BACKGROUND: Destruction of chest wall musculature from trauma, empyema, or local infection limits closure options, especially with muscle flaps. While the vacuum assisted closure system (VAC; KCI International, San Antonio, TX) has been used for wounds in other anatomic locations, we have found no series for chest wounds. METHODS: This is a retrospective review of trauma registry data from the R Adams Cowley Shock Trauma Center, University of Maryland School of Medicine from 2000 to 2003. RESULTS: Seventeen patients were identified and divided into two groups. Group I consisted of seven patients with primary chest wall processes: four necrotizing soft-tissue infections and three with thoracic trauma resulting in significant loss of chest wall musculature. Group II consisted of ten patients with empyema and varying levels of chest wall extension. Six were postpneumonic and four postoperative. Wound size averaged 16 x 7 cm (range, 7 x 3 cm to 21 x 11 cm). The VAC duration averaged nine days (range, 3 to 21 days) and changed every two to three days. Fourteen wounds were culture positive; nine staphylococcus aureus, two alpha hemolytic streptococcus, and one each with enterococcus, Citrobacter, and anaerobes. Eight were polymicrobial. There were no deaths. All wounds healed without rotational muscle flaps. Ten underwent delayed primary closure, four split-thickness skin graft, and three healed by secondary intention. There was one significant complication: a wound infection after delayed primary closure which required reoperation. CONCLUSIONS: Closure of complex chest wall wounds can present significant technical challenges. The VAC system is a simple, useful, and novel alternative to conventional wound care even with large, infected wounds. PMID- 15797048 TI - Adenosine A2A receptor agonist improves cardiac dysfunction from pulmonary ischemia-reperfusion injury. AB - BACKGROUND: Ischemia-reperfusion (IR) injury negatively impacts patient outcome in lung transplantation. Clinically, we observed that lung transplant patients with ischemia-reperfusion injury tend to have cardiac dysfunction. Previous studies have shown that ATL-146e (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), a selective adenosine A2A receptor agonist, reduces lung inflammation after ischemia-reperfusion. We hypothesized that pulmonary ischemia reperfusion causes secondary heart dysfunction and ATL-146e will improve this dysfunction. METHODS: We utilized an in vivo rabbit lung ischemia-reperfusion model. The Sham group underwent 120 minutes single lung ventilation. The IR and ATL groups underwent 90 minutes right lung ischemia with 30 minutes right lung reperfusion. The ATL-146e was given intravenously to the ATL group during reperfusion. Cardiac output and arterial blood gases were monitored, and neutrophil sequestration was measured by myeloperoxidase activity. RESULTS: Upon reperfusion, cardiac output (mL/min) significantly dropped in the IR and ATL groups. By 15 minutes reperfusion, cardiac output in the ATL group improved significantly over the IR group and remained significant thereafter. Lung myeloperoxidase activity was significantly reduced by ATL-146e. Although never hypoxemic, arterial oxygenation was lower in the IR and ATL groups while central venous pressures and mean arterial pressures were similar among groups. A separate experiment demonstrated that reperfusion with the antioxidant N-(2 mercaptopropionyl)glycine prevented cardiac dysfunction. CONCLUSIONS: Pulmonary ischemia-reperfusion causes cardiac dysfunction independent of preload, afterload, and oxygenation. The ATL-146e improves this dysfunction presumably by the antiinflammatory effects of adenosine A2A receptor activation on neutrophils. One likely mechanism involves the release of oxidants from the ischemic lung upon reperfusion, which has immediate negative effects on the heart. PMID- 15797050 TI - Cognitive outcomes three years after coronary artery bypass surgery: a comparison of on-pump coronary artery bypass graft surgery and nonsurgical controls. AB - BACKGROUND: Coronary artery bypass grafting has been associated with both early and late postoperative cognitive decline, but interpretation of previous studies has been limited by lack of appropriate control groups. We compared changes in cognitive performance from baseline to 3 years in patients undergoing coronary artery bypass grafting with those of a control group of patients with known risk factors for coronary artery disease but without surgery. METHODS: Patients undergoing coronary artery bypass grafting (n = 140) and a demographically similar nonsurgical control group with coronary artery disease (n = 92) completed baseline neuropsychological assessment and were followed up prospectively at 3, 12, and 36 months. Cognitive performance was assessed with a battery of neuropsychological tests, measuring the cognitive domains of attention, language, verbal and visual memory, visuospatial, executive function, and psychomotor and motor speed. The statistical analyses were performed in two ways: using data from all tested individuals, and using a model imputing missing observations for individuals lost to follow-up. RESULTS: Both the coronary artery bypass grafting and nonsurgical control groups improved from baseline to 1 year, with additional improvement between 1 and 3 years for some cognitive tests. The coronary artery bypass grafting group had statistically significantly greater improvement than the nonsurgical controls for some subtests, and had a comparable longitudinal course for the remainder of the subtests. Both study groups had a trend toward nonsignificant decline at 3 years on some measures, but the overall differences between groups over time were not statistically significant. CONCLUSIONS: Prospective longitudinal neuropsychological performance of patients with coronary artery bypass grafting did not differ from that of a comparable nonsurgical control group of patients with coronary artery disease at 1 or 3 years after baseline examination. This finding suggests that previously reported late cognitive decline after coronary artery bypass grafting may not be specific to the use of cardiopulmonary bypass, but may also occur in patients with similar risk factors for cardiovascular and cerebrovascular disease. PMID- 15797051 TI - In-hospital patients exposed to clopidogrel before coronary artery bypass graft surgery: a word of caution. AB - BACKGROUND: The aim of our study was to investigate the independent effect of preoperative exposure to aspirin, heparin, and clopidogrel on early clinical outcomes of in-hospital referral patients undergoing first time coronary artery bypass graft (CABG) surgery. METHODS: Outcomes for in-hospital referrals for CABG between January 2001 and January 2002 were prospectively recorded. Exposure to aspirin, heparin, and clopidogrel during the 5 days before surgery was recorded, and their effect on outcome was explored using multiple regression methods. The effect of clopidogrel given within 48 hours before surgery versus stopped at least 48 hours before the operation was also investigated. RESULTS: In-hospital referral patients represented 40% (473 of 1,185) of the total surgical population; 304 (64.7%) were prescribed aspirin, 116 (24.7%) heparin, 91 (19.4%) clopidogrel, and 133 (28.3%) had all anticoagulants stopped 5 days before surgery. In-hospital mortality was 2.3% (95% confidence interval [CI]: 1.1 to 4.1) and was highest among those exposed to clopidogrel (odds ratio 16.9, 95% CI: 2.10 to 136, p = 0.008). Seven of the 11 deaths occurred in patients given clopidogrel. Postoperative intraaortic balloon pump, arrhythmia, reintubation, and stroke were all significantly more common when clopidogrel was prescribed (relative risk estimates all greater than 1.5, p values all < or = 0.021). Blood loss was 37% higher in the patients given clopidogrel (95% CI: +3% to +82%, p = 0.033). Clopidogrel patients were more likely to need a reoperation for bleeding (p < 0.001) and require transfusions (p < or = 0.015). Intubation time and hospital stay were also longer (p = 0.021 and p = 0.019, respectively). Heparin was also associated with longer hospital stay (p < 0.001) and aspirin use with reduced stay (p = 0.041), but for all other outcomes, no association with aspirin or heparin use was found. Comparison of subgroups of clopidogrel patients suggested mortality was highest when clopidogrel was given in the 48 hours before surgery. CONCLUSIONS: Our study suggests that among in-hospital referral patients, preoperative exposure to clopidogrel within 5 days before CABG may increase early mortality and morbidity, and that the mortality risk is greatest when the drug is given within 48 hours of surgery. PMID- 15797052 TI - Noninvasive dynamic assessment with transthoracic echocardiography of a composite arterial Y-graft achieving complete myocardial revascularization. AB - BACKGROUND: Composite arterial grafts are increasingly used in coronary artery bypass surgery. We assessed with transthoracic echocardiography the composite radial artery and in situ left internal thoracic artery Y-graft. METHODS: In 53 of 60 consecutive patients who underwent complete myocardial revascularization using only this composite arterial graft, good transthoracic echocardiographic images and pulsed Doppler signals of the Y-graft main stem were obtained at rest and early after standard exercise. Stress/rest 99mTc-sestamibi myocardial perfusion single-photon emission computed tomography (SPECT) was the gold standard for residual myocardial ischemia. The patients with negative SPECT were divided into groups according to the number of coronary artery systems grafted, and history of preoperative myocardial infarction. RESULTS: Diastolic peak velocity, diastolic velocity-time integral, the diastolic-to-systolic ratio of the peak velocities and velocity-time integrals, and the stress-to-rest ratio of the diastolic peak velocities and diastolic velocity-time integrals in the negative-SPECT patients were significantly greater than in the 6 positive-SPECT patients. Sensitivity and specificity for ischemia of the stress-to-rest ratio of the diastolic peak velocities less than 1.5 were 100%. The stress-to-rest ratio of the diastolic velocity-time integrals in the patients with three coronary systems grafted, and in those without preoperative myocardial infarction, were respectively greater than in the patients with two systems grafted (p < 0.0001), and in those with preoperative myocardial infarction (p = 0.0048). CONCLUSIONS: Noninvasive dynamic assessment with transthoracic echocardiography of a composite arterial graft, including in situ left internal thoracic artery, is feasible and correlates with myocardial perfusion SPECT. The Y-graft used was able to regulate its flow capacity to myocardial demand. PMID- 15797053 TI - Midterm follow-up after minimally invasive direct coronary artery bypass grafting versus percutaneous coronary intervention techniques. AB - BACKGROUND: Revascularization of the left anterior descending coronary artery can be performed by minimally invasive direct coronary artery bypass grafting (MIDCAB) or percutaneous coronary intervention techniques (PCI). The study compared the midterm results of both techniques. METHODS: The outcome of 206 consecutive MIDCAB and 256 PCI patients treated from 1998 until 2001 was retrospectively analyzed. Cardiologists determined the primary patient allocation for the distinct revascularization technique. Periprocedural complications and midterm follow-up, including quality-of-life assessment (SF-36), was reported up to 5.2 years (3.4 +/- 0.7 years). RESULTS: Periprocedural and overall mortality (p = 0.206) showed no differences. Four MIDCAB patients required early reoperation but not for repeated target vessel revascularization. In 16 patients secondary PCI (7.8%) of other coronary vessels was performed. Repeated revascularization of the left anterior descending coronary artery was necessary in 24.2% of patients in the PCI group (p < 0.001), with 4.7% finally requiring surgical revascularization. The incidence of major adverse cardiac events, including myocardial infarction (p = 0.581), repeated target vessel revascularization (p < 0.001), or death (p = 0.206) was higher in the PCI group. This difference consisted basically of the need for repeated target vessel revascularization. Patient-based quality-of-life assessment (SF-36) was independent from the primary chosen revascularization method. CONCLUSIONS: At midterm follow up, MIDCAB resulted in significantly superior results regarding the need for repeated target vessel revascularization compared with PCI, with no significant differences regarding other major cardiac events. PMID- 15797054 TI - Revascularization of the right coronary artery: grafting or percutaneous coronary intervention? AB - BACKGROUND: The choice of myocardial revascularization strategy for the right coronary artery (RCA) in patients with multivessel disease and chronic stable angina remains controversial. Our aim was to determine the better strategy hybrid, combining bypass of the left coronary network and percutaneous coronary intervention of the RCA, or exclusively surgical-and if the latter, the best conduit. METHODS: We used decision analysis, a modeling technique, to compare two RCA revascularization strategies: surgical grafting and percutaneous coronary intervention. A review of the English language literature determined the variables for each strategy. All possible outcomes of each strategy were analyzed to determine the baseline strategy yielding the highest expected effectiveness. Sensitivity analysis determined the most relevant elements in the model and indicated threshold values. RESULTS: Arterial grafting of the RCA led to the highest expected effectiveness, respectively 6% and 7% higher than that of percutaneous coronary intervention and the saphenous graft procedure. Of the arteries available-the radial, right gastroepiploic, and right internal thoracic artery-the most effective was the right internal thoracic artery, pedicled for the proximal part of the RCA and free connected as a Y or a T to the pedicled left internal thoracic artery for the distal part of the RCA. Sensitivity analysis showed surgery to be the appropriate strategy when the expected 1-year patency rate of the arterial graft exceeded 80%. CONCLUSIONS: This analysis shows arterial grafting of the RCA to have better outcomes than percutaneous coronary intervention, and the right internal thoracic artery to be the best conduit. PMID- 15797058 TI - Less-invasive surgical extraction of problematic or infected permanent transvenous pacemaker system. AB - BACKGROUND: The best management of problematic or infected transvenous permanent pacemaker system is complete surgical or percutaneous intravascular extraction of the pacemaker leads and removal of the generator. We present our experiences in 13 such patients in whom the leads were removed with the less-invasive technique. METHODS: From 1996 to 2003, 13 patients, from 31 to 83 years of age (mean, 66.9 +/- 14.0 years), with problematic or infected transvenous permanent pacemaker systems were referred to our department for surgical treatment. In 6 patients, the original pacemakers were dual-chamber. A subxiphoid pericardiotomy was used as the monitoring port during the ventricular lead extraction. In addition, a right parasternal pericardiotomy through the third intercostal space was used as the monitoring port during the atrial lead extraction. RESULTS: Pacemaker systems were completely removed in all patients. Three bleeding episodes (23%), including two right atrial tears and one right ventricular rupture, were successfully circumvented through these monitoring ports. Concomitantly, a new epicardial single-chamber device was implanted through the subxiphoid pericardiotomy whenever indicated in 9 patients. All patients recovered and were discharged uneventfully. At a mean follow-up of 24.8 months (range, 1 to 90 months), no recurrent infections were observed. CONCLUSIONS: A less-invasive technique for explantation of the complete pacemaker system is feasible. This is a reliable method to eradicate infection. Neither cardiopulmonary bypass nor specific intravascular lead extraction devices, such as locking stylets or laser-assisted sheath, are needed. PMID- 15797056 TI - Risk of spinal cord injury after operations of recurrent aneurysms of the descending aorta. AB - BACKGROUND: Degenerative disease of the aorta usually involves the occlusion of several intercostal and lumbar branches by mural thrombus or atherosclerotic plaques, suggesting that the blood supply to the spinal cord is mainly provided through collateral networks. Patients with previous abdominal aortic aneurysm repair and subsequent thoracoabdominal aortic reconstruction must undergo ligation of a number of these segmental arteries, presenting a greater risk of experiencing spinal cord ischemic injury. METHODS: The records of 18 patients who had experienced abdominal aortic aneurysm graft replacement and who had undergone 19 operations for thoracoabdominal aortic repair were retrospectively evaluated. All patients were male. The mean age was 66 +/- 10 years (range, 36 to 75 years); the mean interval between the two operations was 79 +/- 69 months (range, 1 to 231 months). There were 18 (95%) cases of thoracoabdominal aortic aneurysms, and one (5%) case of acute dissection of the thoracoabdominal aorta. The origin of the Adamkiewicz artery was determined preoperatively by computed tomography. Measures to avoid spinal cord injury included monitoring of evoked spinal cord potentials and selective reconstruction of the intercostal arteries under hypothermic cardiopulmonary bypass. RESULTS: There were three (16%) cases of permanent neurologic injury that included one cerebrovascular accident, one neurogenic bladder, and one paraparesis of the right lower limb. There were no cases of paraplegia or postoperative deaths. CONCLUSIONS: Surgical reconstruction of the thoracoabdominal aorta in patients who previously underwent abdominal aortic graft replacement is not related to an increased probability of developing spinal cord ischemic injury. PMID- 15797055 TI - Pulmonary embolectomy for acute massive pulmonary embolism. AB - BACKGROUND: Acute massive pulmonary embolism usually results in death if not diagnosed early and treated aggressively. Thrombolytic therapy and catheter embolectomy are increasingly used as definitive management. Emergent open embolectomy is often reserved as a last resort when less invasive methods have failed or the patient is in cardiopulmonary arrest. METHODS: We reviewed our experience with early open pulmonary embolectomy in patients with acute massive pulmonary embolism from January 1998 to February 2004. RESULTS: Eleven patients underwent early pulmonary embolectomy. Five (45%) patients were men, and the average age was 48 years. In 4 (36%) patients, a massive pulmonary embolism occurred after a surgical procedure or trauma. The remaining 7 patients had chronic medical diseases. The diagnosis was established primarily by clinical findings along with spiral computerized tomography or transesophageal echocardiography. Eight (73%) patients survived and were discharged from the hospital. The 3 patients who died suffered cardiac arrest preoperatively and were taken to the operating room with cardiopulmonary resuscitation in progress. Only 1 patient survived after preoperative cardiac arrest. CONCLUSIONS: Early open pulmonary embolectomy using cardiopulmonary bypass is an effective form of treatment for acute massive pulmonary embolism with excellent long-term results. Preoperative cardiac arrest is associated with a high mortality. Spiral computerized tomography and transesophageal echocardiography are important diagnostic tools. PMID- 15797059 TI - Depression predicts mortality following cardiac valve surgery. AB - BACKGROUND: Depression is associated with mortality in several cardiovascular populations, but has not been evaluated in patients undergoing cardiac valve surgery. Because identifying nonsurgical mediators of survival is important for accurate risk adjustment and the development of interventions to improve outcomes of care, we evaluated the hypothesis that depression predicts mortality following cardiac valve surgery. METHODS: This prospective cohort study enrolled 648 patients undergoing valve surgery at 14 Veteran Administration hospitals. A preoperative mental health inventory (MHI) depression screen was performed in all patients and patients were classified as depressed or not depressed using the standard MHI cutoff score of less than or equal to 52. Multivariable logistic regression was used to evaluate the association between depression and 6-month all-cause mortality, adjusting for other clinical risk variables. RESULTS: Overall, 29.2% (189/648) of the patients were depressed at baseline. Depressed patients were younger, more frequently had New York Heart Association class III/IV symptoms, and more likely required emergent surgery, preoperative intravenous nitroglycerin, or intraaortic balloon pump. Unadjusted 6-month mortality was 13.2% for depressed patients compared with 7.6% for nondepressed patients (p = 0.03). In multivariable analyses, depression remained significantly associated with mortality (odds ratio 1.90; 95% confidence interval 1.07 to 3.40, p = 0.03). These findings were consistent across subgroups of patients undergoing aortic valve replacement, mitral valve replacement and valve replacement without coronary artery bypass graft. CONCLUSIONS: Preoperative depression is an independent risk factor for mortality following cardiac valve surgery. Depression screening should be incorporated into preoperative risk stratification, and future studies are warranted to determine if preoperative or postoperative interventions to treat depression can improve outcomes. PMID- 15797060 TI - Mitral repair versus replacement for ischemic mitral regurgitation. AB - BACKGROUND: We compared mitral repair to replacement in patients with chronic ischemic mitral regurgitation (IMR), due to left ventricular dysfunction (LV-IMR) or papillary muscle infarction (PM-IMR). METHODS: Patients with IMR undergoing repair (n = 65) or replacement (n = 137) from 1990 to 2001 were evaluated. There were 87 patients with LV-IMR, and 115 patients with PM-IMR. Patients presenting in cardiogenic shock were excluded. Outcomes were evaluated by Cox survival analysis with propensity score adjustment and bootstrap validation. RESULTS: Survival at 3, 5, and 9 years was, respectively, 0.94, 0.79, and 0.63 in the repair group, and 0.73, 0.67, and 0.59 in the replacement group. The hazard ratio (HR) of death for mitral repair versus replacement was not constant over the period of follow-up. Repair was associated with better early survival in the PM IMR group, with an adjusted HR of 0.25 (95% confidence interval: 0.09 to 0.71) at 1 year. In the LV-IMR group and in patients with PM-IMR with high acuity and comorbidity, there was no significant survival advantage associated with repair. The beneficial effect of repair was not evident at late follow-up in either group. These findings were independent of the surgeon. Need for reoperation was more common after repair than after replacement (14% versus 3%, p = 0.003). CONCLUSIONS: Patients with PM-IMR benefit from mitral repair with a significantly better early survival. However, the benefit of repair is not evident at longer follow-up. There was a nonsignificant trend toward greater early survival among patients with LV-IMR who underwent repair. PMID- 15797061 TI - An annular prosthesis for the treatment of functional mitral regurgitation: finite element model analysis of a dog bone-shaped ring prosthesis. AB - BACKGROUND: Undersized annuloplasty is commonly used in the treatment of functional mitral regurgitation. However, in the case of severely dilated ventricles, annuloplasty may be inadequate to counteract leaflet tethering. My colleagues and I hypothesized that modifying the shape of the annular prosthesis to account for the specific anatomy of functional mitral regurgitation could challenge extreme leaflet tethering. METHODS: Using finite element model simulations, we tested valve competence after the implantation of conventional D shaped versus dog bone-shaped annuloplasty rings, the latter of which was designed to selectively reduce the septolateral dimension of the annulus. Three models were compared: model A, simulating the native mitral valve; model B, simulating the same valve after annuloplasty with a conventional D-shaped annuloplasty; and model C, simulating a dog-bone annuloplasty ring implantation. Each model was then challenged by progressively pulling the tip of the papillary muscles away from the annulus plane to simulate ventricular remodeling and leaflet tethering. Valve competence was compared in each model for each degree of leaflet tethering. RESULTS: After maximal leaflet tethering simulation (4-mm apical displacement of the papillary tips), the regurgitant area increase was 70.4 mm2 for model A and 52.9 mm2 for model B. In model C, the regurgitant area was only negligibly affected by papillary displacement, increasing to 3.9 mm2. CONCLUSIONS: An annular prosthesis with selective reduction in the septolateral dimension is more effective than a conventional prosthesis for treating leaflet tethering in functional mitral regurgitation. Use of disease-specific annular prostheses is needed to improve the results of valve reconstruction. PMID- 15797063 TI - Comparative evaluation of small-size Sorin Slimline and St. Jude HP heart valve prostheses. AB - BACKGROUND: The Sorin Slimline aortic valve prosthesis, a modification of the Sorin Bicarbon valve with increased internal orifice diameter and geometric orifice area, may show improved hemodynamic performance when compared with other previous prosthesis, but so far no comparison study has been reported. METHODS: Between May 1999 and March 2002, 80 patients (31 to 81 years of age; mean, 65 years) with a small aortic annulus were randomized to undergo aortic valve replacement with either the Sorin Slimline (n = 40) or St. Jude High Performance (n = 40) valve prosthesis. Clinical and echocardiographic Doppler follow-up was performed at 3 to 4 weeks, and 6 and 12 months postoperatively. RESULTS: One patient died of non-valve-related causes 9 months after operation. Mean and peak pressure gradients at 6 and 12 months in the Sorin Slimline valve were lower than in the St. Jude High Performance valve for both size 19 and 21 mm. Effective orifice area and effective orifice area index were not significantly different. There was a significant (p = 0.0001) reduction in left ventricular mass and left ventricular mass index between preoperative measurements and at 12 months after surgery for both valves, but there was no difference (p = 0.27) between the Sorin Slimline and St. Jude High Performance valve prosthesis at any other follow-up period. Clinical results showed similarly good results with both valves. CONCLUSIONS: No clinically significant difference in the hemodynamics of both valves was appreciated; patients with a Sorin Slimline valve exhibited statistically significantly lower pressure gradients, but the small differences in effective orifice area and effective orifice area index did not reach significance. A significant left ventricular mass regression was observed with both valve models. Both prostheses provided a satisfactory clinical outcome. PMID- 15797066 TI - Multiple risk assessment of cardiovascular surgery in chronic renal failure patients. AB - BACKGROUND: Chronic renal failure is a major risk factor in cardiovascular surgery. We evaluated results of cardiovascular surgery in chronic renal failure patients (s-creatinine > 200 micromol/L or established dialysis) at our center from 1990 to 2000. METHODS: One hundred and six chronic renal failure patients underwent cardiovascular surgery (56 coronary artery bypass operations, 25 valve replacements with or without coronary bypass, and 25 other major cardiovascular operations [8 thoracic aorta, 10 abdominal aorta, 7 other]). Matched controls were selected (n = 106) based on age, sex, year, and type of operation and occurrence of diabetes. RESULTS: There were 88 men and 18 women and mean age was 64 +/- 10 years (standard deviation). Demographics did not differ between chronic renal failure and control patients, except for hypertension (more prevalent in chronic renal failure group, p < 0.05). Intraoperative hemorrhage, perfusion and ischemia time, and reoperation did not differ between groups. Chronic renal failure patients received more transfusions of red blood cells, plasma, and platelets (p < 0.02). Ventilation support (27.6 +/- 59.3 hours), intensive care unit stay (7.7 +/- 8.3 days), and hospital stay (12.3 +/- 10.5 days) were longer (p < 0.02). Early mortality was 16% versus 6.6% (p = 0.04) and 5-year mortality was 79% versus 39% (p < 0.05) for chronic renal failure and control patients, respectively. Independent preoperative risk factors of mortality for chronic renal patients were age greater than 70 years (relative risk = 2.32, p = 0.001), chronic obstructive pulmonary disease (relative risk = 2.59, p = 0.001), diabetes (relative risk = 1.80, p = 0.037), and dialysis (relative risk = 2.03, p = 0.005). CONCLUSIONS: Chronic renal failure patients suffered more postoperative complications and had substantially increased short-term and long-term mortality rates. Independent preoperative mortality risk factors for chronic renal failure patients were age, chronic obstructive pulmonary disease, diabetes, and chronic dialysis. PMID- 15797064 TI - Impact of the improvement of valve area achieved with aortic valve replacement on the regression of left ventricular hypertrophy in patients with pure aortic stenosis. AB - BACKGROUND: Previous studies have reported that patient-prosthesis mismatch may be associated with lesser regression of left ventricular hypertrophy. However, among the patients with mismatch, the extent of left ventricular mass regression varied markedly from one patient to another, and we hypothesized that it could be related to the magnitude of the increase in valve area achieved with aortic valve replacement. Our aim was to examine the relationship between the improvement in valve effective orifice area obtained with aortic valve replacement and the extent of postoperative left ventricular mass regression in patients with patient prosthesis mismatch. METHODS: Preoperative and postoperative measurements of valve effective orifice area, transvalvular pressure gradient, and left ventricular mass were obtained by Doppler echocardiography in 111 patients with pure aortic stenosis who were found to have patient-prosthesis mismatch based on an indexed effective orifice area of 0.8 cm2/m2 or less after operation. RESULTS: On average, the valve effective orifice area increased by 0.45 +/- 0.24 cm2/m2 with operation, and mean transvalvular pressure gradients decreased by -31.6 +/- 13.5 mm Hg. The absolute and relative differences between preoperative and postoperative left ventricular mass were -28 +/- 30 g and -17% +/- 18%, respectively. In multivariate analysis, higher preoperative left ventricular mass (p < 0.0001) and larger increase in indexed valve effective orifice area with operation (p = 0.019) were independently associated with greater left ventricular mass regression. CONCLUSIONS: This study shows that in patients with patient prosthesis mismatch, the degree of left ventricular mass regression is influenced by the increase in valve effective orifice area achieved by aortic valve replacement. PMID- 15797062 TI - Midterm outcomes using the physio ring in mitral valve reconstruction: experience in 492 patients. AB - BACKGROUND: Mitral valve reconstruction using standardized Carpentier techniques is the treatment of choice for most patients with regurgitant lesions. Demonstrated predictability and stability make it an attractive alternative to valve replacement. The Physio Ring's inherent flexibility provides a viable alternative in the application of remodeling techniques and appears to be physiologically superior to traditional approaches. METHODS: Between April 1994 and October 2000, 492 consecutive patients underwent mitral valve reconstruction using standardized Carpentier techniques with the Carpentier-Edwards Physio Ring (Edwards Lifesciences LLC, Irvine, CA). There were 267 men (54.3%) and 225 women (45.7%). Mean age was 64.2 years (range, 18 to 86). Almost one-half (44.3%) were 70 years of age or over. The mitral valve etiology was congenital in 7 patients (1.4%), myxomatous in 351 patients (71.3%), ischemic in 88 (17.9%), rheumatic in 26 (5.3%), endocarditis in 9 (1.8%), calcific in 8 (1.6%), and other abnormalities in 3 (0.6%). RESULTS: Isolated mitral valve reconstruction was performed in 282 patients (57.3%), with coronary artery bypass grafting (CABG) in 182 (37.0%), with valve replacement in 11 (2.2%), and with CABG and valve replacement in 17 (3.5%). All patients (100.0%) had ring annuloplasty, 263 (53.5%) leaflet resection, 140 (28.5%) chordal resection, 55 (11.2%) chordal transposition, 48 (9.8%) chordal shortening, and 15 (3.0%) commissurotomy. Overall hospital mortality was 3.5% (17 of 492). Postoperative complications included respiratory insufficiency in 55 patients (11.2%), low cardiac output in 13 (2.6%), stroke in 14 (2.8%), reoperation for bleeding in 13 (2.6%), renal insufficiency in 21 (4.3%), and myocardial infarction in 5 (1.0%), and new onset of atrial fibrillation in 74 patients (15.0%). The cumulative follow-up for the series was 1,522.9 patient years and ranged from 1 to 101.0 months (mean, 38.5 months). There were 11 reconstruction failures (2.3%) requiring ring explant. Actuarial survival was 81.5% +/- 2.1% at 4 years and 67.9% +/- 4.6% at 7 years. Freedom from reoperation at 4 years was 81.5% +/- 2.1% and 67.9% +/- 4.6% at 7 years. CONCLUSIONS: Mitral valve reconstruction with the Physio Ring can be accomplished with low hospital mortality and morbidity even in combined procedures. Moreover, the low incidence of reoperation and late cardiac events suggests that the Physio Ring, with its inherent flexibility, offers a definite advantage in the application of remodeling techniques in mitral valve reconstruction. PMID- 15797067 TI - Early experience with activated recombinant factor VII for intractable hemorrhage after cardiovascular surgery. AB - BACKGROUND: Intractable hemorrhage after complex cardiovascular operations is a serious and potentially lethal complication. We report our experience with the use of activated recombinant factor VIIa (rFVIIa) as rescue therapy for patients with refractory postoperative hemorrhage. METHODS: From April 2002 through December 2003, 9 patients received rFVIIa for intractable hemorrhage after cardiovascular surgery. Patients underwent aortic surgery (2), coronary artery bypass graft surgery (4), double valve operations (2), and mitral valve replacement (1). Four of these procedures were reoperations. Intraoperative aprotinin was used in all patients. All patients underwent standard heparinization (300 IU/kg) before cardiopulmonary bypass and reversal with protamine. RESULTS: Five patients underwent reexploration for mediastinal hemorrhage before treatment; 2 were reexplored twice. The average transfusion requirement before rFVIIa administration was 9 U of blood, 7 U of plasma, 22 U of platelets, and 19 U of cryoprecipitate. rFVIIa was administered as an intravenous bolus at 68 to 120 mug/kg. Mean time of administration from the first operation was 10.9 +/- 7.2 hours. At the time of activated rFVIIa administration, chest tube drainage averaged 640 mL/h. In all patients, chest tube drainage was dramatically reduced to less than 100 mL/h within 5 hours after drug delivery. None of the patients required reexploration after treatment. There were no postoperative neurologic or cardiovascular complications. CONCLUSIONS: When used as rescue therapy for intractable hemorrhage after cardiovascular surgery, rFVIIa may be effective in promoting hemostasis, preventing reexploration, and reducing transfusion requirements. PMID- 15797068 TI - Use of a maze to detect cognitive dysfunction in a porcine model of hypothermic circulatory arrest. AB - BACKGROUND: Hypothermic circulatory arrest (HCA) can result in cognitive impairment not reflected by histopathology or gross neurologic observation. We tested the sensitivity of two multi-room maze tasks in detecting cerebral dysfunction after HCA in pigs. METHODS: Twenty-seven pigs were studied, divided between two tasks. 13 underwent 90 minutes HCA at 20 degrees C and were trained from postoperative day (POD) 7; 14 were unoperated controls. The maze includes a holding area, 8 rooms, and a center hallway. One piece of apple is placed in each baited room on each of 10 days of learning evaluation. After a pig enters a room, doors to all other rooms close, and the pig must return to the holding area. In task 1, 6 of 8 rooms were baited, and each day's session ended when each baited room had been entered, or after 20 trials. In task 2, initially only the right- or left-sided rooms were baited. Pigs were evaluated each day until they entered 4 baited rooms, or for 15 trials; the process was then repeated, baiting the other side. RESULTS: Intraoperative physiology and postoperative recovery showed no differences between task 1 or 2 pigs. Task 1 did not distinguish between control and HCA groups (p = 0.5), but task 2 revealed significantly (p = 0.04) better learning in controls. CONCLUSIONS: The significantly poorer performance of pigs after HCA suggests that the reversal of baited rooms in task 2 provides the sensitivity to detect cognitive dysfunction. The maze is a promising tool to investigate in pigs the mild cerebral damage often seen after HCA. PMID- 15797070 TI - pH-stat versus alpha-stat acid-base management strategy during hypothermic circulatory arrest combined with embolic brain injury. AB - BACKGROUND: There is some evidence of beneficial metabolic effects associated with the pH-stat than with alpha-stat perfusion strategy, but this is tempered by a likely increased risk of embolism to the brain, especially in adult patients. We investigated this possible adverse effect in an experimental model that combined hypothermic circulatory arrest (HCA) and embolic brain injury. METHODS: Twenty-four female juvenile pigs undergoing 25 minutes of HCA at a brain temperature of 18 degrees C were assigned to either alpha-stat (n = 12) or pH stat (n = 12) strategy during cardiopulmonary bypass. Before the initiation of HCA, the descending aorta was clamped and 200 mg of albumin-coated polystyrene microspheres (250 to 750 microm in diameter) were injected into the isolated aortic arch in both groups. RESULTS: The 7-day survival rate was 75% in the pH stat group and 50% in the alpha-stat group (p = 0.40). The pH-stat group had significantly better behavioral scores on postoperative days 5 (p = 0.03) and 6 (p = 0.04). The pH-stat strategy was associated with better postoperative intracranial pressures and histopathologic scores, but such differences did not reach statistical significance. The alpha-stat group had lower brain glucose concentrations postoperatively as well as higher brain lactate/glucose and lactate/pyruvate ratios CONCLUSIONS: These results suggest that pH-stat strategy does not cause any worse brain injury than the alpha-stat strategy. Indeed, the pH-stat strategy is associated with a slightly better outcome compared with the alpha-stat strategy, even in the setting of cerebral embolization. This observation suggests that the pH-stat strategy could also be used in adults during deep hypothermic cardiopulmonary bypass despite the increased risk of intraoperative cerebral embolization. PMID- 15797071 TI - Nafamostat preserves neutrophil deformability and reduces microaggregate formation during simulated extracorporeal circulation. AB - BACKGROUND: Initial sequestration of activated neutrophils and platelet microaggregates in capillaries are responsible for the inflammatory response associated with cardiopulmonary bypass. The study assesses the inhibitory effects of nafamostat mesilate on neutrophil and platelet activation, and on the neutrophil deformability change and microaggregate formation during simulated extracorporeal circulation. METHODS: Fresh heparinized human blood was recirculated for 120 minutes in a membrane oxygenator and a roller pump with and without nafamostat (1.0 mg bolus plus 8.0 mg/h infusion; n = 10 for each group). Neutrophil and platelet counts and platelet aggregation were measured. CD11b, L selectin, and cytoplasmic F-actin of neutrophils were measured by flow cytometry. The microchannel transit time of whole blood was measured as a marker of neutrophil deformability and microaggregate formation. Neutrophil elastase and complement C4d were measured using enzyme immunoassay. RESULTS: Nafamostat preserved platelet counts and inhibited platelet aggregation. Nafamostat significantly reduced neutrophil elastase release at 120 minutes of recirculation, and F-actin expression at 30 and 60 minutes. The drug did not modulate the changes of CD11b, L-selectin, or C4d. Whole blood filterability was significantly preserved by nafamostat at 30 and 120 minutes. CONCLUSIONS: Nafamostat preserves blood filterability during recirculation, possibly by suppression of F-actin expression and platelet activation. Nafamostat may reduce neutrophil sequestration and microaggregate formation in the microcirculation during cardiopulmonary bypass. PMID- 15797072 TI - Is the myofibrillarlytic myocyte a forme fruste apoptotic myocyte? AB - BACKGROUND: Myofibrillarlytic (MFL) cells are commonly observed in subendocardial myocardium in myocardial infarction. Because ischemic damage to myocytes is also known to induce apoptosis, we evaluated the prevalence of apoptosis in MFL cells in nine ischemic cardiomyopathic hearts explanted during transplantation. METHODS: Myocytes with partial or complete clearing of cytoplasm, observed commonly in the subendocardium, were recognized as MFL cells. Prevalence of apoptosis was defined by TUNEL and ISOL staining and further characterized by immunohistochemical staining for caspase-3, Bcl2, BCL-X(L), Bax, proliferating cell nuclear antigen (PCNA), and Ki67. RESULTS: Of 4131 MFL cells examined, 1305 (32%) possessed nuclei in a given histologic section; 1140 (88%) of the nucleated myocardial cells were TUNEL positive. Of 842 cells with normal appearance, 257 (31%) cells demonstrated nuclei in the given histologic section. TUNEL staining was observed in 5 (1.9%) in these control areas. All MFL cells stained positive for caspase 3. The antiapoptotic proteins, Bcl2 and BCL-X(L), demonstrated intense upregulation within and surrounding MFL cells, whereas pro-apoptotic protein Bax expression was only seen at control level. The MFL cells had Ki67 negative and PCNA positive nuclei. CONCLUSIONS: The present study demonstrates that the majority of MFL cells are apoptotic and are associated with upregulation of caspase 3. Simultaneous upregulation of Bcl2 represents a survival effort in these myocytes. This is consistent with the review of the literature that MFL cells are viable, persist in myocardium for long time and may be functionally reversible. Evidence for concurrent apoptosis and survival instinct represent a conceptual paradox and suggests that myocytes undergoing apoptosis should be amenable to reconstitution of function. PMID- 15797074 TI - Principal strain orientation in the normal human left ventricle. AB - BACKGROUND: Methods that can improve the accuracy of application of directed intervention in the treatment of coronary artery disease deserve investigation. Magnetic resonance imaging with tissue tagging allows for noninvasive, quantitative determination of regionally varying minimum principal strain. Because the directional vector of minimum principal strain has been shown to be sensitive to ischemic involvement, my colleagues and I sought to fully characterize the normal range of vector direction in the in vivo human left ventricle at rest and during inotropic stimulation. METHODS: Tagged magnetic resonance imaging image sets were acquired in 20 healthy volunteers at rest and during dobutamine infusion. Strain was computed from the measured displacement data by using finite element software. Orientation of minimum principal strain was characterized by measuring the angle (principal strain angle) between the minimum principal strain vector and the local circumferential-longitudinal plane. Values of this angle were computed in 6 ventricular regions and globally. RESULTS: Resting values of the principal strain angle were small in every region, confirming that maximal normal myocardial contraction occurs primarily in the circumferential-longitudinal plane. Angles were similar during dobutamine infusion. Comparisons between ventricular walls, both at rest and with dobutamine, revealed no marked regional differences in the principal strain angle. CONCLUSIONS: The direction of maximal myocardial contraction is known to change with ischemic injury to the myocardium and can be a sensitive, regionally varying index of myocardial ischemia. The critical first step in the clinical application of this technology is to accurately characterize normal ranges of principal strain angles. PMID- 15797075 TI - Multislice computed tomography for preoperative evaluation of right ventricular volumes and function: comparison with magnetic resonance imaging. AB - BACKGROUND: This study was performed to validate preoperative right ventricular measurements obtained from multislice spiral computed tomography data sets in comparison with magnetic resonance imaging. METHODS: Before cardiac surgery, 25 patients (among them 12 patients with compromised right ventricular function) underwent contrast-enhanced retrospectively electrocardiogram-gated multislice spiral computed tomography and cine magnetic resonance imaging in a standardized fashion. Right ventricular end-diastolic, end-systolic and stroke volume, ejection fraction, and myocardial mass were calculated according to the slice summation method. Measurements obtained with both modalities were compared using Pearson's correlation coefficient (r), Student's t test for paired samples, and Bland-Altman analysis. RESULTS: The right ventricle was completely visualized with invariably adequate image quality on all multislice spiral computed tomography and magnetic resonance images. For all measurements a close correlation between multislice spiral computed tomography and magnetic resonance imaging was found (end-diastolic volume, r = 0.93; end-systolic volume, r = 0.95; stroke volume, r = 0.91; ejection fraction, r = 0.96; mass, r = 0.94). Mean values of all measurements did not differ significantly between both modalities, and limits of agreement were in an acceptable range. CONCLUSIONS: When compared with magnetic resonance imaging as a reference method, multislice spiral computed tomography seems to be an accurate and reliable noninvasive technique for evaluating right ventricular measurements. PMID- 15797076 TI - Sternal nourishment in various conditions of vascularization. AB - BACKGROUND: Early changes in sternal perfusion were studied after midline sternotomy and different methods of mammary artery (MA) harvesting. METHODS: Our observations were made in the swine model after midline sternotomy. In group 1 (6 animals), after unilateral skeletonized MA harvesting, (99m)Tc particles were injected intravenously. In group 2 (7 animals), after unilateral mammary artery and vein harvesting (semiskeletonized technique), (99m)Tc particles were injected intravenously. In group 3 (5 animals), after skeletonized bilateral MA harvesting, 99mTc particles were injected into the intercostal musculature lateral to the sternal border. In groups 1 to 3, sternal samples were analyzed using gamma counting. In group 4 (6 animals), unilateral skeletonized MA harvesting was performed. In group 5 (5 animals), the MA was harvested unilaterally using the semiskeletonized technique. In groups 4 and 5, sternal blood flow was assessed using thermography. Data were collected in all groups for 5 hours postoperatively. RESULTS: Both radioactive and thermographic flow measurements showed a statistically significant decrease in sternal blood flow on the side of harvested mammary vessels, regardless of harvesting technique. Radioactivity of the devascularized hemisterni on the side of intramuscular particle injection was substantially higher than in the contralateral half, confirming the role of diffusion in sternal nourishment. The distal sternal segments were least perfused by the MA. CONCLUSIONS: There is an acute reduction of sternal perfusion during the early postoperative period, even if collaterals are preserved by skeletonized MA harvesting. Diffusion plays an important role in sternal nourishment, particularly of the xiphoid, and even more so after MA harvesting. PMID- 15797077 TI - Operative results and outcomes in children with Shone's anomaly. AB - BACKGROUND: The outcome of children with multilevel left heart obstructions with mitral valve involvement (Shone's anomaly) is generally poor. This study reviews our results and outcomes in patients with staged repair for left ventricular outflow tract obstruction (LVOTO) and mitral valve reconstruction. METHODS: A total of 27 children underwent staged repair for Shone's anomaly between 1978 and 2003. The mean age at the first operation was 9.0 months (range, 2 days to 3 years). Mitral stenosis was present in all; with supravalvular mitral ring (n = 11), mitral valve abnormalities including parachute mitral valve, fused chordae, single papillary muscle (n = 10), and "typical" (Ruckman & Van Praagh) congenital mitral stenosis (n = 14). The LVOT obstruction features included subaortic stenosis (n = 16), valvar aortic stenosis (n = 7), bicuspid aortic valve (n = 24), and coarctation (n = 18). All 27 patients underwent 94 surgical procedures, including 22 mitral operations and 48 LVOT operations. RESULTS: There were no operative deaths at the first operation. Mean follow-up was 4.7 +/- 4.4 years (range, 6 months to 15 years). There were three late deaths (11%). All late deaths were secondary to severe mitral valve disease. Overall 15-year actuarial survival was 89%. All surviving patients are in New York Heart Association functional class I or II. CONCLUSIONS: Patients with Shone's anomaly are seen with a wide spectrum of major anatomic and hemodynamic abnormalities. A favorable outcome is possible for most. Operative mortality is adversely affected by the severity of mitral valve disease, the degree of left ventricular hypoplasia, and the need for multiple operative procedures. PMID- 15797078 TI - Aortic valve prolapse associated with outlet-type ventricular septal defect. AB - BACKGROUND: Aortic valve prolapse is frequently associated with juxta-arterial ventricular septal defect. The significance of its association with other outlet types of ventricular septal defect, however, remains unclear. METHODS: From 1987 to 2002, 677 patients (male:female ratio, 424:253) who received surgical repair for ventricular septal defect extending to the outlet septum were reviewed. Based on surgical findings, ventricular septal defects were classified as juxta arterial, perimembranous outlet, or muscular outlet type. RESULTS: Aortic valve prolapse occurred in 373 of 677 patients (57.2%) with 209 juxta-arterial, 103 perimembranous outlet, and 61 muscular outlet type. Significant aortic regurgitation developed in 51 of 373 (14%). Among 252 patients with regular follow-up, the mean onset ages of aortic valve prolapse in juxta-arterial, perimembranous outlet, and muscular outlet type were 4.9, 5.0, and 5.1 years, respectively (no statistical difference). The presence of larger shunt and probably anterior malalignment predicted an earlier onset of aortic valve prolapse. Perimembranous outlet and muscular outlet type ventricular septal defect were frequently associated with infundibular hypertrophy and subaortic ridge, and perimembranous outlet type was associated with anterior septal malalignment. In juxta-arterial ventricular septal defect and ventricular septal defect with anterior malalignment, prolapsed cusp was always the right coronary cusp, but noncoronary cusp involvement was also common in perimembranous outlet type (17 of 103, 16.5%). CONCLUSIONS: The association with anterior septal malalignment, infundibular stenosis and subaortic ridge is related to the location of the outlet ventricular septal defect. The age of onset of aortic valve prolapse in each type was quite similar, and a larger shunt may predict an earlier onset. PMID- 15797080 TI - Robotic mitral valve annuloplasty with double-arm nitinol U-clips. AB - PURPOSE: Robotic mitral valve repair increases precision however operative times are longer. Prior studies have indicated that robotic knot tying is time consuming and it is without potential room for improvement. We therefore investigated tissue approximation devices that may shorten operative times. DESCRIPTION: A 67-year-old female was approached through a right mini-thoracotomy with the da Vinci Robotic Surgical System (Intuitive Surgical, Sunnyvale, CA). Using 12 nitinol U-clips (Coalescent Surgical, Sunnyvale, CA) an annuloplasty band was placed under robotic guidance. Clip placement and deployment times were recorded and statistical comparisons were assessed to prior suture annuloplasties. EVALUATION: Clip placement time was 1.3 +/- 0.9 (minutes +/- standard deviation), statistical comparison with first, most recent, and all prior suture annuloplasties proving no significance. Clip deployment time was 0.5 +/- 0.2, whereas knot-tying times and respective statistical comparison for first, most recent, and all prior suture annuloplasties were 2.0 +/- 0.7 (p = 0.003), 1.2 +/- 0.4 (p = 0.0004), and 1.6 +/- 0.6 (p < 0.00001). Follow-up echocardiography performed postoperatively, at 3 months, and at 9 months revealed valvular structural integrity with only minimal mitral regurgitation. CONCLUSIONS: U-clips considerably reduce time for annuloplasty over conventional suture and may help reduce operative times as well. PMID- 15797082 TI - MagScrew total artificial heart in vivo performance above 200 beats per minute. AB - PURPOSE: Downsizing pulsatile devices requires an increase of beat rate if flow capacity is to be maintained. We applied this concept to the preclinical MagScrew total artificial heart (TAH). DESCRIPTION: The device fills passively with a stroke volume of 45 ml and beat rates up to 250 beats per minute (bpm). EVALUATION: Stable hemodynamics were observed during a 30-day bovine implant with a flow of 8.7 +/- 1.2 L/min at beat rates of 204 +/- 18 bpm. Device filling was exceptional up to 250 bpm generating flow of greater than 12 L/min. Beat rate adapted to preload in a way similar to a Frank-Starling response. Left and right atrial pressures were balanced. The aortic pulse pressure was 49-70 mm Hg, which translates to a pulsatility index of 0.49-0.77. Organ functions were preserved and blood damage did not occur. CONCLUSIONS: Increasing the beat rate while downsizing the MagScrew TAH was successful with strong flow generation by passive filling. Pulsatility was maintained at high beat rates. This innovative approach may be used to develop small pulsatile pumps. PMID- 15797084 TI - A new method of temporary epicardial atrioventricular pacing utilizing bipolar pacing leads. AB - PURPOSE: We evaluated a convenient method of temporary atrioventricular pacing utilizing bipolar epicardial pacing leads that offer better sensing and pacing performance. DESCRIPTION: Fifty-one patients undergoing coronary artery bypass grafting had atrial and ventricular bipolar leads implanted. The ventricular leads were inserted onto the front of the right ventricle, and the atrial leads were inserted into the lateral muscular part of the right atrium near the interatrial groove. Sensing values, pacing thresholds, and impedance were measured on all leads on postoperative days 0, 2, and 4, and complications of insertion and removal were noted. EVALUATION: The method was convenient and there were no complications during insertion or removal. The mean pacing threshold increased from 1.1 V to 1.5 V in both the atrial and ventricular leads from day 0 to day 4 (not significant). The mean sensed p wave amplitude decreased from 2.2 mv to 2.0 mv (not significant), and the mean sensed R wave amplitude decreased from 6.2 mv to 4.1 mv (p = 0.001) from day 0 to day 4. In spite of this significant drop in the sensed R wave amplitude, this value remained in an acceptable range. There was also a significant decrease in impedance, but overall all values were in an acceptable range assuring safe and effective pacemaker function. Only one atrial lead (2%) and three ventricular leads (6.1%) failed to pace on day 4. CONCLUSIONS: We conclude that this temporary epicardial pacing method is safe, convenient, and less time consuming. Satisfactory pacing and sensing performance was achieved with low thresholds and minimal complications. PMID- 15797086 TI - Heparin-induced thrombocytopenia associated with bilateral adrenal hemorrhage after coronary artery bypass surgery. AB - Heparin-induced thrombocytopenia (HIT) is a well-recognized syndrome associated with thrombosis and multiple potential clinical sequelae. We report a case of bilateral adrenal hemorrhage, a known but rare complication of heparin-induced thrombocytopenia complicating a routine coronary artery bypass surgery. Thrombocytopenia, abdominal pain, and signs of adrenal insufficiency in the context of heparin treatment should raise suspicion of this unusual complication. PMID- 15797087 TI - Psoriasis-induced postoperative cardiac failure. AB - Erythroderma-induced cardiogenic shock is a rare but known manifestation of erythrodermic psoriasis. Erythrodermic psoriasis is an aggressive form of the disease and can be precipitated by numerous factors including stress to the body. In this article we present a case of persistent severe vasodilatation after coronary artery bypass surgery that required prolonged epinephrine administration. This was caused by a flareup of erythrodermic psoriasis that was previously only controlled by methotrexate. This drug was omitted in the perioperative phase. Vasodilatation disappeared after treatment was reinstituted. PMID- 15797088 TI - Left atrial appendage aneurysm. AB - Aneurysms of the left atrial appendage are exceedingly rare. Patients most commonly present with atrial tachyarrhythmias and thromboembolism. Resection of the aneurysm is usually curative. We present a case report of a 60-year-old female with a long-standing history of atrial arrhythmias found to have a large left atrial appendage aneurysm with additional bi-atrial enlargement and a family history of atrial arrhythmias. The patient was successfully treated with resection of the aneurysm and a Cox-Maze III procedure. PMID- 15797089 TI - Aortic valve-sparing procedure with cusp elongation and free edge reinforcement for bicuspid aortic valve. AB - Aortic valve-sparing procedures demonstrate excellent valvular function at midterm. Recently authors have reported acceptable early results with aortic valve-sparing procedures on patients with regurgitant bicuspid valves. We report the case of a novel procedure to preserve bicuspid valves with a calcified raphe and root dilatation. This procedure includes raphe excision, pericardial patch elongation, free edge leaflet reinforcement, and a root reimplantation valve sparing procedure. PMID- 15797090 TI - Reoperation of the Kay-Shiley disc valve 35 years after replacement. AB - The Kay-Shiley disc valve (Shiley Inc, Irvine, CA) was manufactured in 1965 and is no longer in clinical use due to its high incidence of thromboembolism. We report a case of tricuspid valve replacement with the Kay-Shiley valve 35 years previously. The valve was replaced successfully with a St. Jude Medical valve (St. Jude Medical, Inc, St. Paul, MN). This is the longest interval from implantation to re-replacement with the Kay-Shiley valve that has been reported in the literature. PMID- 15797091 TI - Stentless bioprosthetic aortic valve replacement after a homograft root replacement: Toronto SPV implantation after a homograft root. AB - Stentless bioprosthetic valves for the aortic position offer excellent hemodynamic characteristics, making them an attractive choice ahead of other valve prostheses. We present a unique case in which a patient underwent aortic valve replacement with a stentless porcine valve and mitral valve repair for severe aortic and mitral regurgitation 1 year after a homograft root replacement for acute aortic endocarditis. The rationale for our approach is outlined in the context of current surgical trends. PMID- 15797092 TI - Isolated unilateral absence of right proximal pulmonary artery: surgical repair and follow-up. AB - The isolated unilateral absence of a proximal pulmonary artery is a rare congenital lesion with a diverse clinical presentation. If the connecting ductus arteriosus closes after birth, the ipsilateral pulmonary artery will lose its source of blood supply, resulting in hypoplasia or obliteration of intrapulmonary vessels. Despite a seemingly benign early clinical course, a significant number of untreated patients will develop pulmonary hypertension, hemoptysis, and recurrent respiratory infections. Early detection and surgical repair provides restoration of physiologic pulmonary circulation, regression of pulmonary hypertension, and the potential for normal distal pulmonary vascular development. PMID- 15797093 TI - Epitheloid hemangioendothelioma of the heart in infancy. AB - We report a case of primary cardiac epithelioid hemangioendothelioma arising from the right atrium of a 2-month-old infant. The tumor was found incidentally during exploratory sternotomy for recurrent pericardial effusion. This case represents a very rare situation, because this is the youngest patient found in relevant literature, and because it involves extensive infiltration by the tumor without any development of intracardiac mass appearance. PMID- 15797094 TI - Sternocleidomastoid myocutaneous esophagoplasty for benign cervical stricture. AB - Benign strictures of the cervical esophagus that are resistant to dilation present a formidable challenge to the surgeon. Numerous varied techniques have been developed to restore swallowing. Reports of the sternocleidomastoid myocutaneous pedicled flap for repair of benign cervical strictures are scarce. We are reporting a case of residual lye stricture that was treated with sternocleidomastoid myocutaneous esophagoplasty in order to promote awareness of this procedure. This one-stage operation took 1 hour to complete. The patient began eating on postoperative day 4 and has not had any further symptoms. PMID- 15797095 TI - Primary pulmonary meningioma first suspected of being a lung metastasis. AB - Primary extracranial and extraspinal meningiomas are rare tumors. We describe a primary pulmonary meningioma first suspected of being a metastasis because it presented as a solitary subpleural pulmonary nodule in a patient with breast cancer. The absence of radiographic change after 6 months of chemotherapy led to resection of the breast and lung lesions. A complete central nervous system evaluation eliminated other locations of meningioma, allowing the diagnosis of primary pulmonary meningioma. PMID- 15797096 TI - A direct measurement of serum amylase levels produced by lung cancer. AB - The production of amylase by lung cancer has been previously diagnosed pathologically or immunohistochemically, or it has been confirmed by a decrease in serum levels after resection. It is possible to directly probe the continuous production of amylase by collecting samples from the inflow and outflow vessels of lung cancer. We herein describe an intraoperative measurement in which the amylase level in the pulmonary lobar vein was 3 times greater than that in the superior vena cava which was 6 times greater than the normal range. The venous level of amylase returned to a normal range after resection. PMID- 15797097 TI - Posterior intrathoracic neurinoma cure: a transforaminal resection after a thoracotomy. AB - Posterior intrathoracic neurogenic tumors often have an intraforaminal extension. Ten percent have an extension intraspinal and are called "dumbbell tumors." The surgically recommended techniques used to treat these conditions may have been applied too systematically, mainly when the diameter of the foramen was obviously enlarged and the possibility of removing the tumor existed. We report two cases to stress the importance of assessing the size of the intervertebral foramina in planning neurinoma cure. PMID- 15797098 TI - A rare inferior middle mediastinal tumor resection under extra-corporeal circulation. AB - An inferior middle mediastinal mass was observed in a 51-year-old woman initially presenting as pericardial effusion and mimicking lymph node disease. Thoracotomy demonstrated a tumor invading the posterior atrial wall. Complete resection was possible under the extracorporeal circulation. The tumor proved to be a soft tissue myoepithelioma in an exceptional location. Despite the difficult surgical resection due to location, prognosis of this absolutely rare tumor without cytologic features of malignancy should be good. PMID- 15797099 TI - Multiple pulmonary nodules and underlying head and neck cancer. AB - We describe a 66-year-old woman with recently diagnosed cT2N0 mouth cancer and multiple hypo-dense pulmonary nodules discovered on a computed tomographic chest scan. These nodules were located in the anterior part of the right upper and middle lobe and were resected thoracoscopically. Histologic examination of these nodules revealed a lipoid pneumonia. Exogenous lipoid pneumonia is a rare but described pulmonary disease that typically presents as consolidations. An exclusive presentation such as multiple pulmonary nodules is very unusual. PMID- 15797100 TI - Massive recurrent anterior diaphragmatic hernia, coronary artery disease, and valvular heart disease. AB - We report the case of a 76-year-old woman found on preoperative evaluation for vaginal prolapse to have coronary artery disease, aortic stenosis, and a large recurrent anterior diaphragmatic hernia. The clinical presentation and surgical management of this case, and the review of the literature are discussed. PMID- 15797101 TI - The longest functioning apical left ventricular to descending aortic valve conduit. PMID- 15797102 TI - Large organized parietal thrombus of the right ventricle after aortic valve prosthetic implantation 5 years previously. PMID- 15797103 TI - The "eaves" technique for distal anastomosis in aortic arch replacement. AB - We describe the "eaves" technique, a new method for distal anastomosis in aortic arch replacement. The 1-cm wide eaves were created at the site 3 to 4 cm distal to the graft end. The graft was bound with vessel tape from the eaves to the site proximal to the origin of the first branch to make a working space above the eaves and to facilitate graft handling. Then the native descending aorta was sutured to the eaves easily. The eaves enabled a greater surface contact area between the graft and the inner wall of the aorta and reduced bleeding at the anastomosis. PMID- 15797104 TI - Using fascia lata to treat infective aortic false aneurysm. AB - In patients with infective aortic false aneurysms, repair using artificial materials is in danger of becoming an additional focus for infection. We used harvested autologous fascia lata as a vascular patch in such operations on 2 patients with infected mediastinal false aneurysm after coronary artery bypass surgery. These patients have not had any recurrences of mediastinitis and false aneurysm for 4 to 6 years after the operations. PMID- 15797105 TI - Transposition of the great arteries and quadricuspid pulmonary valve: an unusual combination. AB - Quadricuspid semilunar valves are very rare, especially in association with other congenital cardiac anomalies. A quadricuspid pulmonary valve has never been described in the setting of transposition of the great arteries. In this brief article we describe one such case, and we discuss the operative strategy during the arterial switch operation with particular reference to the technique of coronary translocation. PMID- 15797106 TI - A new, easy method for putting "U" stitches inside the chest wall. AB - In this report, we describe a new, easy method for putting "U" stitches inside the chest wall. The method does not require extension of the skin incision nor subcutaneous dissection and it minimizes chest wall injury. This method may also be applied to other surgical fields where needles can penetrate the wall of the cavity when it is difficult to stitch from the inside of the cavity. PMID- 15797108 TI - Animal models of heart failure: what is new? AB - Heart failure is the major cause of mortality in Western countries. Medical treatment of heart failure is associated with 50% survival at 5 years. Experimental models are required to better understand the progression of the disease and elaborate new therapy. Heart transplantation, left ventricular assist devices, artificial hearts, and cardiac bioassist techniques require animal models for testing and optimizing before they are implemented on human patients. The perfect model of heart failure that reproduces every aspect of the natural disease does not exist. Acute and chronic heart failure models have been developed to reproduce different aspect of the pathology. PMID- 15797107 TI - In the footsteps of senning: lessons learned from atrial repair of transposition of the great arteries. AB - The Senning operation has evolved from being the initial surgical correction that allowed survival in complete transposition of the great arteries to an integral part of the anatomic repair of congenitally corrected transposition. In patients with complete transposition, the Senning operation has given satisfactory initial and long-term surgical results, but the potential for right ventricular failure and atrial arrhythmias have drastically reduced its indications in the current era. The long-term follow-up and pertinent postoperative issues of the Senning operation will be reviewed, along with its newfound role in the anatomic repair of congenitally corrected transposition. PMID- 15797109 TI - The Society of Thoracic Surgeons practice guideline series: aspirin and other antiplatelet agents during operative coronary revascularization (executive summary). PMID- 15797111 TI - Do off-pump surgical procedures really improve memory? PMID- 15797113 TI - Pulmonary artery banding still has an important role in the treatment of congenital heart disease. PMID- 15797114 TI - Hemoptysis. PMID- 15797116 TI - Endothelialization and functional neointima on vascular grafts in humans. PMID- 15797118 TI - Neointima in vascular prostheses: the jury is still out. PMID- 15797119 TI - Pneumonectomy versus sleeve lobectomy for lung cancer. PMID- 15797121 TI - Creatine kinase. PMID- 15797122 TI - One-pot acetalation-acetylation of sugar derivatives employing perchloric acid immobilised on silica. AB - Perchloric acid immobilised on silica gel has been used as an efficient promoter for per-O-acetylation, and acetalation and subsequent O-acetylation of glycosides and thioglycosides in one-pot using stoichiometric reagents. PMID- 15797123 TI - Bicyclic nucleoside analogues from D-glucose: synthesis of chiral as well as racemic 1,4-dioxepane ring-fused derivatives. AB - The dioxepanofuranose derivatives 4 and 12, obtained through the cyclization of the 3-(2-hydroxyethyl) ether of a D-xylo-pentodialdose derivative, were appropriately functionalized and elaborated to the first examples of the new class of 3'-O and 5'-O-bicyclic nucleoside analogues 9, 10, and 14 with a fused seven-membered ring. Reactions carried out through the intermediacy of the D-xylo pentodialdose derivative 5 yielded racemic products, while prior protection of the 4-formyl group (as in 7) before deprotection of the 1,2-hydroxyl groups led to optically active analogues. PMID- 15797124 TI - Production of a new sucrose derivative by transglycosylation of recombinant Sulfolobus shibatae beta-glycosidase. AB - The gene encoding beta-glycosidase of the hyperthermophilic archaea Sulfolobus shibatae (SSG) was expressed in Escherichia coli. Recombinant SSG (referred to as rSSG hereafter) was efficiently purified, and its transglycosylation activity was tested with lactose as a donor and various sugars as acceptors. When sucrose was used as an acceptor, we found a distinct intermolecular transglycosylation product and confirmed its presence by TLC and high performance anion exchange chromatography (HPAEC). The sucrose transglycosylation product was isolated by paper chromatography, and its chemical structure was determined by 1H and 13C NMR. The sucrose transfer product was determined to be beta-D-galactopyranosyl-(1 ->6)-alpha-D-glucopyranosyl-beta-d-fructofuranoside with a galactose molecule linked to sucrose via a beta-(1-->6)-glycosidic bond. PMID- 15797125 TI - Revised structures for the capsular polysaccharides from Staphylococcus aureus Types 5 and 8, components of novel glycoconjugate vaccines. AB - Glycoconjugate vaccines based on the capsular polysaccharides (CPSs) from Staphylococcus aureus serotypes 5 and 8 conjugated to genetically detoxified recombinant exoprotein A (rEPA) from Pseudomonas aeruginosa have been shown, in Phase 3 clinical trials, to elicit a strong bactericidal immune response in end stage renal disease patients. Such vaccines have the potential to reduce morbidity and mortality due to methicillin-resistant Staphylococcus aureus (MRSA), a major cause of hospital-acquired infection. The serotype 5 and 8 polysaccharides have been fully characterized by NMR spectroscopy and full structural analyses carried out. Published structures were found incorrect and the revised structures of the repeat units of the two polysaccharides are: [carbohydrate structure: see text]. Resonances indicative of the presence of peptidoglycan were observed in the spectra of both CPSs, consistent with reports that the CPS is covalently linked to peptidoglycan. PMID- 15797126 TI - Structural characterisation of a highly branched exopolysaccharide produced by Lactobacillus delbrueckii subsp. bulgaricus NCFB2074. AB - Lactobacillus delbrueckii subsp. bulgaricus NCFB2074 when grown in skimmed milk secretes a highly branched exopolysaccharide. The exopolysaccharide has a heptasaccharide repeat unit and is composed of glucose and galactose in the molar ratio 3:4. Using chemical techniques and 1D and 2D NMR spectroscopy the polysaccharide has been shown to possess the following repeat unit structure: [carbohydrate structure: see text]. PMID- 15797127 TI - The structure of kappa/iota-hybrid carrageenans II. Coil-helix transition as a function of chain composition. AB - This paper describes the effect of the kappa/iota-ratio on the physical properties of kappa/iota-hybrid carrageenans (synonyms: kappa-2, kappa-2, weak kappa, weak gelling kappa). To this end, a series of kappa/iota-hybrid carrageenans ranging from almost homopolymeric kappa-carrageenan (98 mol-% kappa units) to almost homopolymeric-carrageenan (99 mol-% iota-units) have been extracted from selected species of marine red algae (Rhodophyta). The kappa/iota ratio of these kappa/iota-hybrids was determined by NMR spectroscopy. Their rheological properties were determined by small deformation oscillatory rheology. The gel strength (storage modulus, G') of the kappa/iota-hybrids decreases with decreasing kappa-content. On the other hand, the gelation temperature of the kappa-rich kappa/iota-hybrids is independent of their composition. This allows one to control the gel strength independent of the gelation or melting temperature. The conformational order-disorder transition of the kappa/iota hybrids was studied using optical rotation and high-sensitivity differential scanning calorimetry. High-sensitivity DSC showed that the total transition enthalpy of the kappa/iota-hybrids goes through a minimum at 60 mol-% kappa units, whereas for the mixture of kappa- and iota-carrageenan, the total transition enthalpy is a linear function of the composition. With respect to the ordering capability, the kappa/iota-hybrid carrageenans seem to behave as random block copolymers with length sequence distributions truncated from the side of the small lengths. Intrinsic thermodynamic properties (e.g., transition temperature and enthalpy) of kappa- and iota-sequences in these copolymers are close to those of their parent homopolymers. The critical sequence length for kappa-sequences is 2-fold of that for iota-sequences. PMID- 15797128 TI - Chemical characterization of the immunomodulating polysaccharide of Aloe vera L. AB - The polysaccharide isolated by alcohol precipitation of Aloe vera mucilaginous gel was found to have a Man:Glc:Gal:GalA:Fuc:Ara:Xyl ratio of 120:9:6:3:2:2:1 with traces of Rha and GlcA. Linkage analysis of the endo-(1-->4)-beta-d mannanase-treated sample yielded Manp-(1--> (approximately 26%), 4-Manp (approximately 53%), 2,4-Manp (approximately 3%), 3,4-Manp (approximately 1%), 4,6-Manp (approximately 1%), 4-Glcp (approximately 5%), 4-Xylp (approximately 1%), Xylp-(1--> (approximately 2%), Galp-(1--> (approximately 5%), and traces of 4,6-Galp and 3,6-Galp. Hydrolysis with strong acids produced a mixture of short oligosaccharides and an acid-resistant fraction containing greater relative fractions of Manp-(1-->, Araf-(1-->, Xylp-(1-->, and 4-Xylp than the bulk polysaccharide. NMR analysis of oligosaccharides generated by endo-(1-->4)-beta-D mannanase and acid hydrolysis showed the presence of di-, tri-, and tetrasaccharides of 4-beta-Manp, beta-Glcp-(1-->4)-Man, beta-Glcp-(1-->4)-beta Manp-(1-->4)-Man, and beta-Manp-(1-->4)-[alpha-Galp-(1-->6)]-Man, consistent with a backbone containing alternating -->4)-beta-Manp-(1--> and -->4)-beta-Glcp-(1--> residues in a approximately 15:1 ratio. Analysis of the sample treated sequentially with endo-(1-->4)-beta-d-mannanase and alpha-D-galactosidase showed that the majority of alpha-Galp-(1--> residues were linked to O-2, O-3, or O-6 of -->4)-beta-Manp-(1--> residues, with approximately 16 -->4)-beta-Manp-(1--> residues between side chains. Our data provide direct evidence of a previously proposed glucomannan backbone, but draw into question previously proposed side chain structures. PMID- 15797129 TI - Study on the synergetic degradation of chitosan with ultraviolet light and hydrogen peroxide. AB - Chitosan was effectively degraded by hydrogen peroxide under irradiation with ultraviolet light. The existence of a synergetic effect on the degradation was demonstrated by means of viscometry. In addition, the optimal conditions of degradation were determined on the basis of orthogonal tests. The structure of the degraded product was characterized by Fourier-transform infrared spectra (FTIR) analysis and diffuse reflectance spectra (DRS) analysis. The mechanism of the degradation of chitosan was correlated with cleavage of the glycosidic bond. PMID- 15797130 TI - Structural analysis of carrageenans from the red alga, Callophyllis hombroniana Mont. Kutz (Kallymeniaceae, Rhodophyta). AB - The use of a range of modern analytical techniques has facilitated the structural characterisation of the polysaccharide from the New Zealand endemic red alga, Callophyllis hombroniana. The native polysaccharide contains a number of structural units with the largest proportion consisting of 3-linked beta-D galactopyranosyl 2-sulfate units, alternating with 4-linked 3,6-anhydro-alpha-D galactopyranosyl 2-sulfate units, that is, theta-carrageenan (36 mol%). C. hombroniana is the first red seaweed reported to naturally contain such a large proportion of theta-carrageenan. PMID- 15797131 TI - Structural characterization of beta-D-(1-->3)-glucans from different growth phases of the marine diatoms Chaetoceros mulleri and Thalassiosira weissflogii. AB - We have investigated the content and structure of the chrysolaminarans isolated from the two marine diatoms Chaetoceros mulleri and Thalassiosira weissflogii. Samples were taken from different phases of growth, and the structure of the chrysolaminaran was seen in relation to the specific growth rate of the diatoms. The structure determined for the glucan from C. mulleri was found not to vary with different specific growth rates. T. weissflogii showed some variance in the structure, both throughout the different stages of growth and between samples taken from the stationary phase. C. mulleri was found to have a chrysolaminaran with a degree of polymerization (DP) of 22-24 and a degree of beta-(1-->6) branching of 0.006-0.009. These results corresponded well with previous results obtained in our laboratories. The chrysolaminaran isolated from T. weissflogii was found to have a DP of 5-13 and no beta-(1-->6) branching. This is to our knowledge the first characterization of the chrysolaminaran from T. weissflogii. PMID- 15797132 TI - Determination of the three-dimensional structure of oligosaccharides in the solid state from experimental 13C NMR data and ab initio chemical shift surfaces. AB - A novel method for the determination of the three-dimensional (3D) structure of oligosaccharides in the solid state using experimental 13C NMR data is presented. The approach employs this information, combined with 13C chemical shift surfaces (CSSs) for the glycosidic bond carbons in the generation of NMR pseudopotential energy functions suitable for use as constraints in molecular modeling simulations. Application of the method to trehalose, cellobiose, and cellotetraose produces 3D models that agree remarkably well with the reported X ray structures, with phi and psi dihedral angles that are within 10 degrees from the ones observed in the crystals. The usefulness of the approach is further demonstrated in the determination of the 3D structure of the cellohexaose, an hexasaccharide for which no X-ray data has been reported, as well as in the generation of accurate structural models for cellulose II and amylose V6. PMID- 15797133 TI - Crystal structure and solid state 13C NMR analysis of nitrophenyl 2,3,4,6-tetra-O acetyl-beta-D-gluco- and D-galactopyranosides. AB - The X-ray diffraction analysis of o-nitrophenyl 2,3,4,6-tetra-O-acetyl-beta-D galactopyranoside (1), m-nitrophenyl 2,3,4,6-tetra-O-acetyl-beta-D galactopyranoside, p-nitrophenyl 2,3,4,6-tetra-O-acetyl-beta-D-galactopyranoside and o-nitrophenyl 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranoside was performed. It was found that except in the case of 1, all other crystals have one molecule in the independent part of the crystal unit cell. The results support the opinion that the nitro group does not conjugate effectively with the phenyl ring. In the 13C CP MAS spectrum of 1 the signals are split, confirming the presence of two independent molecules. Similarly, the 13C CP MAS NMR spectrum of p-nitrophenyl 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranoside indicated the presence of two non equivalent molecules in the crystal unit. One of these molecules has more conformational freedom enabling rotation of the phenyl ring. PMID- 15797134 TI - A new radiochemical method to investigate ion binding with polyelectrolytes. AB - A new method for investigating the binding of ions with polyelectrolytes has been developed. This method, based on Donnan equilibrium and an isotope exchange between the electrolyte and polyelectrolyte, can distinguish territorial from specific binding of ions and can determine fractions of ions bound with the polyion. This method can determine ion binding with polyelectrolytes in a wide range of polyelectrolyte concentrations in multicomponent solutions. The method was tested with radioactive tracers 22Na+, 36Cl- and heparin sodium salt. The influence of the ionic strength on the Na+ binding with heparin was investigated at 310 K. In the limit of zero ionic strength, all Na+ ions are bound to heparin, but only 45% of them are exchangeable. Thus Na+ ions can be bound both territorially and specifically. The fraction of bound ions decreases rapidly with increasing ionic strength. The fraction of the specifically bound ions becomes negligible when the ionic strength exceeds 0.01 M, whereas the fraction of territorially bound ions can be neglected at ionic strengths higher than 0.45 M. PMID- 15797135 TI - An investigation of pectin methylesterification patterns by two independent methods: capillary electrophoresis and polysaccharide analysis using carbohydrate gel electrophoresis. AB - The analysis of partially methylesterified oligogalacturonides plays a key role both in the elucidation of the fine structure of the polysaccharide pectin and in the study of pectin-acting enzymes. Experimental methods performing the separation, detection and quantification of oligogalacturonides are, therefore, of crucial importance in the drive to understand structure-function relationships in pectin containing systems, both in vitro and in vivo. In this work standard samples of unesterified and partially methylesterifed galacturonides, and enzymatic digests of several pectin samples possessing distinct intramolecular patterns of methylesterification were studied using capillary electrophoresis (CE) and polysaccharide analysis using carbohydrate gel electrophoresis (PACE). In addition to yielding interesting information regarding the fine structures of the different pectic substrates digested, the study has been used as a vehicle in order to compare the two recently reported methods. PMID- 15797136 TI - A fluorescence study of sodium hyaluronate/surfactant interactions in aqueous media. AB - The interactions between sodium hyaluronate, an anionic polysaccharide, with surfactants (anionic and nonionic) were investigated using pyrene fluorescence measurement methods. The change of micropolarity produced by the interaction was monitored by the measurement of emission intensity ratio between the first and third bands (I1/I3), and the intensity ratio of the excimer and the third vibration monomer band (I(E)/I(M)). Because the hydrophilic heads on the SDS were attracted by the domains formed by the hydroxyl groups of hyaluronate, the I1/I3 ratio was reduced by the addition of hyaluronate at lower than 0.06% of sodium dodecyl sulfate (SDS) concentration. No aggregation was observed between hyaluronate and nonionic surfactants (Tween-80 and Cremophor EL) in the whole concentration range studied. At a higher concentration of surfactant, the I1/I3 ratio of hyaluronate/surfactant was influenced by the addition of saccharide (glucose, lactose, or mannitol). However, the effect of saccharide could be reduced by the addition of salt. PMID- 15797137 TI - Octanol-water partition coefficient of glucose, sucrose, and trehalose. AB - The octanol-water partition coefficients (P) of glucose, sucrose, and trehalose were measured at temperatures between 5 and 20 degrees C using an enzymatic method. The measured log P is compared with calculated and experimental data previously reported. In the case of trehalose, the measured log P differs considerably from the theoretically estimated values and agrees with the expected value for a disaccharide. Some methods of assessing the partition coefficients are also analyzed and it is concluded that the atom/fragment contribution method overestimates the hydrophilicity of disaccharides and, probably in a larger extension, that of trisaccharides. The knowledge of P for these sugars is valuable both for basic or applied purposes, including food and biomolecules stabilization. PMID- 15797138 TI - An expedient synthesis of benzyl 2,3,4-tri-O-benzyl-beta-D-glucopyranoside and benzyl 2,3,4-tri-O-benzyl-beta-D-mannopyranoside. AB - An efficient three-step synthesis of benzyl 2,3,4-tri-O-benzyl-beta-D glucopyranoside, a widely used building block in carbohydrate chemistry, is described. The key step is the selective debenzylation-acetylation of perbenzylated beta-glucose using ZnCl2-Ac2O-HOAc. This approach was also used to affect an efficient three-step synthesis of benzyl 2,3,4-tri-O-benzyl-beta-D mannopyranoside. PMID- 15797140 TI - Synthesis of regioselectively and uniformly modified maltoheptaose derivatives from cyclomaltoheptaose precursors. AB - Heptadeoxy-6(I-VII)-halo, -azido, and hepta-6(I-VII)S-hepta(N-Boc-2-amino)ethyl 6(I-VII)-heptathiomaltoheptaose derivatives were prepared by acetolysis of the corresponding per-C-6-modified beta-cyclodextrin derivatives. The rapid and convenient structural characterisation of all of the modified oligosaccharides by ESIMS is described. PMID- 15797139 TI - 9-O-Sulfation on alpha-NeuAc-(2-->8)-NeuAc and inter-residue lactonization. AB - Treatment of alpha-NeuAc-(2-->8)-NeuAc (1) with SO3-pyridine (4 equiv) in DMF resulted in selective 9-O-sulfation on the nonreducing end residue and the formation of an inter-residual delta-lactone. The lactonization could result from the C-2 carboxylic acid of the nonreducing residue condensing with the hydroxyl group or/and sulfated group at C-9 of the reducing residue to form a six-membered ring between two adjacent sialic acid residues. When alpha-NeuAc-(2-->9)-NeuAc (5) was used as a sulfation substrate, only 9-O-sulfation on the nonreducing end residue was observed. According to capillary electrophoresis (CE) analysis, 9-O sulfation on the disialic acid is a fast reaction, while sulfation on other hydroxyl groups is insignificant under the conditions used. PMID- 15797141 TI - Enzymatic synthesis of oligosaccharides from branched cyclodextrins. AB - 6''-alpha-Maltosyl-maltotriose and 6''-alpha-D-glucosyl-maltotriose were prepared from Novamyl degradation of 6-O-alpha-maltosyl-alpha-cyclodextrin and 6-O-alpha-D glucosyl-alpha-cyclodextrin, respectively. NMR spectroscopy was used to elucidate their structural identities, in a combination of COSY experiments. Further, a mechanism for the degradation was proposed based on the Novamyl active site geometry. PMID- 15797142 TI - Chitooligosaccharides--preparation with the aid of pectinase isozyme from Aspergillus niger and their antibacterial activity. AB - An isozyme of pectinase from Aspergillus niger with polygalacturonase activity caused chitosanolysis at pH 3.5, resulting in low-molecular weight chitosan (86%), chitooligosaccharides (COs, 4.8%) and monomers (2.2%). HPLC showed the presence of COs with DP ranging from 2 to 6. Charcoal-Celite chromatography and re-N-acetylation of the COs followed by CD, IR, MALDI-TOF-MS and FAB-MS analyses revealed an abundance of chitobiose, chitotriose and chitotetraose. The COs monomeric mixture showed a bactericidal effect towards Bacillus cereus and Escherichia coli more efficiently than native chitosan. Among the chitooligomers, the hexamer showed maximum antibacterial effect followed by the penta-, tetra-, tri- and dimers. Of the two monomers, only GlcN showed slight bacterial growth inhibition. SEM revealed bactericidal action patterns of COs-monomeric mixture towards B. cereus and E. coli. PMID- 15797143 TI - A new neutral polymer from the cell wall of actinomycete Kineosporia aurantiaca VKM Ac-702T. AB - The major cell wall polymer of Kineosporia aurantiaca VKM Ac-702T a representative of the suborder Frankineae, is a galactomannan with a repeating unit of the following structure: -->3)-beta-D-Galp-(1-->6)-beta-D-Manp-(1-->4) beta-D-Manp-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Manp-(1-->4)-beta-D-Manp-(1--> that has not been reported so far. This was established using chemical degradation methods and NMR spectroscopy. The polysaccharide identified in the present study provides the second example of neutral galactomannans in actinomycete cell walls. The cell wall of K. aurantiaca VKM Ac-702T also contains a minor teichoic acid, viz., 1,3-poly(glycerol phosphate) partially substituted with alpha-glucosamine residues, only part of which are N-acetylated. PMID- 15797144 TI - Structural analysis of the core oligosaccharide from Pasteurella multocida strain X73. AB - The structure of the core oligosaccharide region of the lipopolysaccharide from the Pasteurella multocida strain X73 was elucidated. The lipopolysaccharide was subjected to a variety of degradative procedures. The structure of the purified oligosaccharide was established by monosaccharide and methylation analyses, NMR spectroscopy and mass spectrometry. The following structure illustrates a similar structure to the recently identified oligosaccharide from another P. multocida strain VP161, but with additional symmetrical substitution of the terminal galactose residues with phosphoethanolamine moieties, where based on the NMR data all sugars were found in pyranose ring forms and Kdo is 3-deoxy-alpha-D-manno-2 oct-2-ulosonic acid, l,D-alpha-Hep is l-glycero-D-manno-heptose, PEtn is phosphoethanolamine and PCho is phosphocholine. PMID- 15797145 TI - Structure of the O-polysaccharide from the Azospirillum lipoferum Sp59b lipopolysaccharide. AB - A neutral O-specific polysaccharide was obtained by mild acid hydrolysis of the lipopolysaccharide of the plant-growth-promoting bacterium Azospirillum lipoferum Sp59b. On the basis of sugar and methylation analyses along with 1D and 2D (1)H and (13)C NMR spectroscopy, including a NOESY experiment, the following structure of the branched hexasaccharide repeating unit of the O-polysaccharide was established: [carbohydrate structure: see text]. PMID- 15797146 TI - Creative writing in health care: a branch of complementary medicine. PMID- 15797147 TI - Nurse administered telephone intervention for blood pressure control: a patient tailored multifactorial intervention. AB - OBJECTIVES: A randomized controlled trial involving a nurse administered patient tailored intervention is being conducted to improve blood pressure (BP) control. METHODS: Veterans with hypertension from an outpatient primary care clinic completed a baseline assessment and were randomly allocated to either a nurse administered intervention or to usual care. In this ongoing study, intervention patients receive the tailored intervention bi-monthly for 2 years via telephone; the goal of the intervention is to promote adherence with medication and improve health behaviors. Patient factors targeted for intervention include perceived risk of hypertension, memory, literacy, social support, patients' relationship with their health care provider, side effects of therapy, pill refill, missed appointments, and health behaviors. RESULTS: The sample randomized to the nurse intervention consisted of 294 veterans with hypertension (average age = 63 years; 41% African-American). A comparable sample of veterans was assigned to usual care (n = 294). We have maintained a 97% retention rate for the first 12 months of the study. The average phone call has lasted 3.7 min ranging from less than 1 to 40 min. At 6-month post-enrollment, individuals receiving the nurse intervention had a greater increase in confidence with following hypertension treatment (P < 0.007) than the usual care group. DISCUSSION: The intervention is easily implemented and is designed to enhance adherence with prescribed hypertension regimen. The study includes both general and patient-tailored information based upon need assessment. The study design ensures internal validity as well as the ability to generalize study findings to the clinic settings. PMID- 15797148 TI - Factors influencing breastfeeding practices and postponement of solid food to prevent allergic disease in high-risk children: results from an explorative study. AB - This paper presents results of seven focus group interviews conducted to gain insight into the feelings, opinions and perceived barriers of parents with a history of asthma who have recently delivered a child. The parents participated in an educational program regarding breastfeeding and postponement of solid food to prevent their child from developing allergic symptoms. Breastfeeding exclusively for 6 months seemed an advice difficult to follow. The most important influencing factors regarding initiation and continuation of breastfeeding were health advantages for the baby, bonding, social support, modelling, knowledge about all the aspects of breastfeeding and breastfeeding confidence. In general, parents adhered to the advice to postpone solid food until the child had reached the age of 6 years. The few obstacles revealed were social pressure, hungry babies and eagerness of parents to give solid food. PMID- 15797149 TI - Changes in physician-patient communication from initial to return visits: a prospective study in a haematology outpatient clinic. AB - Limited research has investigated how physician-patient interaction changes over time. We have therefore examined physician-patient communication during the two initial, as well as the seventh (on average) patient visit to a haematology outpatient clinic. Consultations were audio taped and analyzed using the Roter interaction analysis system (RIAS). Patients completed the Impact of Events Scale (IES) before and a satisfaction questionnaire after each consultation. Consultations were generally physician dominated and task-focused. While the amount of task-focused communication was significantly reduced between the initial and the return visits, the amount of socio-emotional communication remained quite stable. In return visits (but not in the two initial visits), patients with more severe diagnoses were given longer consultations and they provided more task-focused information to a less verbally dominant physician. Patients were more satisfied in the second and return visits (but not in the first), if consultations contained greater levels of socio-emotional communication. PMID- 15797150 TI - Prevalence and correlates of perceived unmet needs of people with systemic lupus erythematosus. AB - To assess the prevalence and correlates of perceived unmet need among people with systemic lupus erythematosus (SLE) 386 people with SLE, recruited from a support association in NSW, Australia, completed a mail-back survey that included 97 items measuring perceived unmet needs across seven domains: physical, daily living, psychological/spiritual/existential, health services, health information, social support and employment/financial. Ninety-four percent of participants had at least one unmet need. The highest levels of unmet need were tiredness (81%); pain (73%); not being able to do things one used to (72%); fear of exacerbation (72%); sleeping problems (70%); anxiety and stress (69%); and feeling down (68%). Five of the highest levels of unmet needs were in the psychological domain. Researchers concluded that current health care delivery is not adequately meeting some of the needs of people with SLE. A comprehensive needs assessment questionnaire for all SLE patients could help providers manage protocols more effectively. PMID- 15797152 TI - Father's health in relation to spousal employment. AB - We studied the correlation between fathers' well-being and the number of hours that their wives work outside the home. Two groups of fathers were involved in the study: modern, caring fathers (in this cultural context meaning less than full-time employment outside the home) and more traditional fathers (in full-time employment). It would appear to be true for both groups that the number of hours their wives work correlates negatively with the fathers' well-being both physically and mentally. We compared two hypotheses: does the well-being of fathers correlate mainly with the wife's actual absence, measured by the absolute number of hours that she works outside the home? Or does it correlate more with symbolic factors, measured by the relative number of hours she works (her working hours compared with his), or by her relative share to the family income. Confirmation was found for both hypotheses, but the second, the 'unfulfilled husband' hypothesis had the most support. Finally, it must be observed that the number of hours men work still has a positive correlation with their well-being. PMID- 15797151 TI - Variation in perceived competence, glycemic control, and patient satisfaction: relationship to autonomy support from physicians. AB - There is considerable variation in care provided to patients with diabetes related to metabolic control, preventive services, and degree of patient-centered support. This study evaluates the relation of self-determination theory (SDT) constructs of clinician autonomy support, and patient competence to glycemic control, depressive symptoms, and patient satisfaction from baseline surveys of 634 patients of 31 Colorado primary care physicians participating in a program to improve diabetes care. Spearman correlations of autonomy support from one's clinician with patient competence, HbA1c, depressive symptoms and satisfaction were significant (R = -0.11 to 0.55, P < 0.005). Structural equation modeling demonstrated that autonomy support was significantly related to perceived competence, depressive symptoms, patient satisfaction, and indirectly to glycemic control. Perceived competence was significantly related to depressive symptoms, patient satisfaction and glycemic control. Further, the motivation constructs from SDT accounted for 5% of the variance in glycemic control, 8% of the variance in depression, and 42% of the variance in patient satisfaction. Quality improvement efforts need to pay greater attention to patient competence, satisfaction, and depression, in addition to glycemic control. Clinician autonomy support was found to be reliably measured and moderately correlated with psychosocial and biologic outcomes related to diabetes self-management. These results suggest training clinicians to increase their support of patient autonomy may be one important avenue to improve diabetes outcomes. PMID- 15797153 TI - Patient participation in discussing palliative radiotherapy. AB - Cancer patients' participation in doctor-patient interactions has been shown to be an important factor in the emotional processing of their condition, particularly when only palliative treatments can be offered. In this study, we assessed incurable cancer patients' participation in initial consultations with their radiation oncologists (ROs). RO stimulation of patient participation and discussions about treatment decisions were also measured. The entire consultation was videotaped and analyzed using the Roter Interaction Analysis System (RIAS). Patients' participation proved to be low on medical information, but high on discussing their experiences and life circumstances. The ROs stimulated patient participation mainly by providing medical information and giving patients opportunities to tell their stories. Decisions about radiation treatment had previously taken place and were rarely discussed in the consultations studied. The results suggest that patient participation in palliative treatment consultations might be improved for facilitating patients' emotional processing of the incurable nature of their cancer. PMID- 15797154 TI - Encouraging patient adherence: primary care physicians' use of verbal compliance gaining strategies in medical interviews. AB - Compliance-gaining strategies refer to subtle differences in ways people use language when their goal is to influence someone else's behavior. This stands in contrast to other kinds of persuasion aimed only at influencing others beliefs and attitudes. We have developed a new method of coding what physicians say when they are trying to influence patients' behaviors. This method applies theory and methods from the fields of interpersonal influence, linguistics and social psychology. We tested the reliability of this new method by randomly selecting 37 audiotaped medical interviews collected for an unrelated study [J. Gen. Int. Med., 9 (1994) 402] and having three coders independently identify physician compliance-gaining utterances and then independently apply one of 57 codes to each utterance. These codes also were categorized on two underlying dimensions reflecting whether the physician (1) framed the compliance-gaining utterance in a direct or indirect way, and (2) did or did not give a justification for that direct or indirect request. Reliability among coders and coders agreement with the final utterance identification and coding decisions, measured as per cent agreement among coders and/or, where appropriate, by Cohen's kappa were good to excellent. Most physicians' strategies were indirect and incomplete. For female patients, physicians used significantly more strategies, including more indirect strategies, complete strategies, "prescriptions" and "demands". For male patients, physicians used a greater percent of direct strategies, including "procedural demands". This method provides a reliable and promising new technique for observing naturally occurring physician compliance-gaining speech. PMID- 15797156 TI - Evaluation of a decision aid for making choices about intubation and mechanical ventilation in chronic obstructive pulmonary disease. AB - To assist patients with chronic obstructive pulmonary disease (COPD) in advance planning for life-threatening exacerbations, we developed a structured decision aid that describes the process, risks, and outcomes of intubation and mechanical ventilation (MV). Thirty-three patients with severe COPD took part in a before after evaluation study. At baseline, only two participants (6%) reported that they had already made an advance decision about MV. After reviewing the decision aid, 31 participants (94%) reported that they had made a choice, which in 23 cases (74% of those deciding) was to forego MV. These choices were associated with more accurate expectations of MV outcome, and reduced decisional conflict. Qualitatively, participants who would accept MV emphasized their wish to prolong life, whereas those who would forego MV were more influenced by the burdens of treatment and the perception of a poor long-term outcome. However, there was evidence that 24% of participants did not completely comprehend the decision aid and 27% found the experience to be stressful. These findings indicate that a decision aid for MV helps patients plan for life-threatening exacerbations, and may be a useful adjunct to counseling for some patients with severe COPD. PMID- 15797155 TI - Patients' interest in discussing cancer risk and risk management with primary care physicians. AB - Little is known about patients' preferences for discussing cancer risks and risk management with primary care physicians. We sought to determine whether patients want to discuss such topics and what factors are associated with this interest. Participants (375 patients ages 40-85, of diverse race and education level) completed a telephone survey prior to scheduled physician visits. Survey included items on perceived health, perceived cancer risk, education level, and whether participants would like to discuss with a physician their breast, ovarian or colon cancer risk, tamoxifen, cancer genetic counseling, and colon cancer screening. Greater proportions were interested in discussing risks for each cancer, compared with those who were not (P < 0.0001). More participants were interested in discussing mammograms (80%) and cancer genetic counseling (60%) than tamoxifen (49%) or colon cancer screening modalities (43-53%). For many topics, poorer perceived health was associated with greater interest in future discussion; higher education level was associated with less interest. PMID- 15797157 TI - Attention deficit and hyperactivity disorder/learning disabilities (ADHD/LD): parental characterization and perception. AB - Sixty-six parents of adolescents (mean age, 14.8 years), who attended special education classes and who were diagnosed as having attention deficit and hyperactivity disorder/learning disabilities (ADHD/LD), were interviewed. The comorbidity of the ADHD group included emotional lability and/or depression, 70%; oppositional defiant disorder (ODD), 67%; obsessive-compulsive disorder (OCD), 44%; addiction to buying, 44%; and aggressiveness, 62%. Twenty-one percent were either involved in the past or presently using drugs. Nine percent had attempted suicide. According to their parents, the main characteristic of these adolescents was low self-image. Parents enumerated five negative characteristics: impulsiveness; nervousness; angered easily ('short fused'); aggressiveness with cursing and outbursts; and impaired sociability with impoliteness. PMID- 15797158 TI - A short educational intervention on communication skills improves the quality of screening for Chlamydia in GPs in Belgium: a cluster randomised controlled trial. AB - An accurate algorithm for screening for chlamydial infections is available in general practice, but GPs experience numerous barriers to sexually transmitted infections (STI) counselling. In this study we assessed if a short educational package, under the form of a commented video footage on communication skills, was helpful in implementing the screening strategy. A cluster randomised controlled trial was carried out in 36 general practitioners in Antwerp, Belgium. Main outcome measures were: number of patients included in the risk assessment, number of patients tested, and proportion of appropriately tested patients. The results show that GPs in the intervention group did not include more patients overall, but that the quality of the screening process was significantly better (81.6% versus 56.2% appropriate tests, P = 0.02). Conclusively, GPs who participated in a short educational package on communication skills, selected eligible candidates for screening more accurately and decreased the risk of overscreening. PMID- 15797159 TI - The effect of two types of memory training on subjective and objective memory performance in healthy individuals aged 55 years and older: a randomized controlled trial. AB - The objective of the study was to examine the effectiveness of two types of memory training (collective and individual), compared to control (waiting list), on memory performance. Participants were 139 community-dwelling older individuals recruited through media advertisements asking for people with subjective memory complaints to participate in a study. Data were collected at baseline, and at 1 week and 4 months after the intervention. Training efficacy was assessed using measures of subjective and objective memory performance. After the intervention, participants in the collective training group reported more stability in memory functioning and had fewer feelings of anxiety and stress about memory functioning. In addition, positive effects were found on objective memory functioning. Compared with the other two groups, the collective training group participants had an improved recall of a previously learned word list. Compared to controls, participants in the individual training group reported fewer feelings of anxiety and stress in relation to memory functioning. PMID- 15797160 TI - Using expert patients' narratives as an educational resource. AB - In this article, we examine the topic of patient participation in health care and report on how we have drawn on the concept of patient expertise to produce a new kind of information booklet for chronic arthritis patients. The booklet is patient-generated and contains the illness narratives of patients with three kinds of arthritis. The booklet draws upon the knowledge of patients who feel they are flourishing despite their condition. By communicating information through the powerful medium of narrative it is hoped the booklet will be a useful educational and supportive resource for other patients with a similar condition. We would encourage health professionals to acknowledge patient expertise and to consider the expertise of certain patients as a valuable educational resource both for themselves and for other patients. PMID- 15797161 TI - Parents' and medical personnel's beliefs about infant teething. AB - Many symptoms are attributed to teething. Little evidence exists to support these beliefs, despite their implications on clinical management. This study attempted to investigate parental and medical personnel's beliefs about teething. The study was conducted by means of a questionnaire, submitted to 55 pediatricians and 130 nurses. A parent survey was conducted simultaneously. We evaluated 462 questionnaires. An association of teeth eruption with infant morbidity was believed to exist by 76% of the responders, mostly by parents and nurses, and less so by physicians. Irritability, fever, and loose stools/diarrhea were believed to be the most common symptoms associated to teeth eruption. We conclude that almost all parents, the majority of nurses, and many physicians believe that teething is associated with the appearance of symptoms, most of which are minor and relate to discomfort rather than physical illness, but a substantial minority still ascribes potentially serious symptoms to teething. PMID- 15797162 TI - Willingness of pulmonologists to guide COPD patients in smoking cessation. AB - Effective smoking cessation interventions include steps often protocolised as the 4A method. This study assessed how pulmonologists address the smoking behaviour of chronic obstructive pulmonary disease (COPD) patients and analysed psychosocial differences between pulmonologists who intend to use the 4A method and those who do not intend to use it. A nationwide survey was conducted among Dutch pulmonologists (N = 320), of whom 63% responded. Pulmonologists routinely assessed patients' smoking behaviour and their willingness to quit, but were least experienced in actually assisting patients to quit. Pulmonologists intending to use the method perceived a more supportive social environment in working with it and were more convinced of their ability to use it. Overall, pulmonologists were not convinced that the method would result in more patients quitting, and were pessimistic about their ability to use it. Pulmonologists need to be convinced of the effectiveness of the 4A method and their ability to use it. Skills training and the creation of a social environment supportive toward smoking cessation assistance are recommended. PMID- 15797163 TI - Improving adherence to a cholesterol-lowering diet: a behavioral intervention study. AB - Less than 50% of US adults follow dietary recommendations. Despite these figures, little research has focused on improving adherence to a therapeutic eating plan. The research utilizing self-efficacy theory has shown promise for improving behavior change and treatment adherence. This study evaluated the efficacy of a telephone-delivered, self-efficacy based intervention designed to improve adherence to a cholesterol-lowering diet among those self-reporting nonadherence. Sixty-five men and women diagnosed with hypercholesterolemia were randomized to usual care or treatment, which consisted of six intervention sessions delivered every 2 weeks by telephone and focused on how to manage eating behavior in challenging situations. There were significant between group differences post intervention in the consumption of saturated fat (P < .001) and cholesterol (P = .040) with the intervention group improving their dietary adherence. Significant change (P = .013) occurred over time in low-density lipoprotein-cholesterol (LDL C) in the intervention group. No changes were observed in self-efficacy between groups, suggesting that self-efficacy was not a mediator of the improved adherence. The study's findings confirm that the telephone is a useful tool to deliver adherence-enhancing interventions. PMID- 15797164 TI - Learning through connections with others: women's cardiac symptoms. AB - Conducting group sessions in which women can share experiences, develop connections, and learn from each other may potentially be a powerful health educational tool. In a focus group conducted for evaluative feedback, the authors found that women began spontaneously sharing their experiences and feelings, receiving validation, and learning from each other almost from the start of the group session. The participants who were dissimilar in age, ethnicity/race, and socioeconomic status developed a strong connection based on the common experience of having had a heart attack. Although the participants had received patient education after their heart attack, they learned new information about cardiac symptoms and how to respond to them in the group session. This case example is a compelling reminder that women learn through connections. Therefore, a group approach to health education that acknowledges the salience of personal experiences, the experiences of others, the development of connections, and the exchange of information may provide a meaningful learning environment. Further research needs to be conducted to determine if conducting group appointments for women that fosters the development of connections and the exchange of information enriches the learning environment and enhances patient education. PMID- 15797165 TI - Regression-based formulas for predicting change in RBANS subtests with older adults. AB - Repeated neuropsychological assessments are common with older adults, and the determination of clinically significant change across time is an important issue. Regression-based prediction formulas have been utilized with other patient and healthy control samples to predict follow-up test performance based on initial performance and demographic variables. Comparisons between predicted and observed follow-up performances can assist clinicians in making the determination of change in the individual patient. The current study developed regression-based prediction equations for the twelve subtests of the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) in a sample of 223 community dwelling older adults. All algorithms included both initial test performances and demographic variables. These algorithms were then validated on a separate elderly sample (n = 222). Minimal differences were present between Observed and Predicted follow-up scores in the Validation sample, suggesting that the prediction formulas would be useful for practitioners who assess older adults. A case example is presented that utilizes the formulas. PMID- 15797166 TI - Test-retest reliability of the Extended Complex Figure Test--Motor Independent administration (ECFT-MI). AB - The Extended Complex Figure Test (ECFT; Fastenau, 1996b, 2003a), adds recognition and matching trials to the Rey-Osterrieth Complex Figure Test (ROCFT). An alternate administration of the ECFT, the ECFT-Motor Independent version (ECFT MI; Fastenau, 2003a) measures visuospatial memory without the motoric demands of the ROCFT. ECFT-MI test-retest reliabilities were examined in 55 healthy adults tested two times separated by one week. The Recognition Trial showed strong reliability for the Total Score (r = .80, p < .0001). Reliabilities for Recognition Detail Scale and Subscales (r = .62-.74, p < .0001) and for the Global Scale (r = .51, p < or = .005) were attenuated by fewer items per subscale but were comparable to other memory tests. Matching Trial reliability coefficients were attenuated by extreme ceiling effects and restriction of range, but scores were very consistent between the two administrations. This study supports the temporal stability of the ECFT-MI. PMID- 15797167 TI - Comparison of various RAVLT scores in the detection of noncredible memory performance. AB - Sixty-one noncredible patients (as documented by psychometric and behavioral criteria) scored significantly below 25 controls and 88 credible clinic patients with no motive to feign on most RAVLT scores, including added indices of implicit and "automatic" memory. A combination of true recognition (i.e., recognition minus false positives) + implicit memory score (i.e., the number of word stems completed with RAVLT items) + "automatic" memory score (i.e., the number of correct temporal order judgments) < or =22 was associated with 75.7% sensitivity with specificity at 91.5%. However, sensitivity was nearly as high when scores available from the standard RAVLT administration alone (i.e., no word stem or temporal order trials) were combined. Specifically, a cut-off of < or =12 for true recognition (recognition minus false positives) + primacy recognition (i.e., number of words recognized from the first third of the test) was associated with 73.8% sensitivity at 90% specificity. These results indicate that combined indices of recognition memory from the RAVLT are effective in identifying noncredible memory performance in "real world" samples and are modestly superior to the 67.2% sensitivity obtained with the standard recognition score. PMID- 15797168 TI - A comparison of methods for measuring cognitive change in older adults. AB - Well-researched statistical methods are required to guide clinicians in determining the significance of test score changes in serial neuropsychological assessment of older adults. The following six change score methods were examined using five-year test-retest data from the Canadian Study of Health and Aging: the standard deviation method, three reliable change indices (RCIs), and two standardized regression-based methods. Changes in scores on four memory measures were examined in cognitively healthy older adults, and the RCI with a correction for practice/aging effects most accurately classified this normal variability. Diagnostic change (i.e., developing dementia versus remaining cognitive healthy) was also examined in relation to memory test score changes. All change score methods were significantly associated with diagnostic change, though the strength of association varied by measure and method. In contrast to some previous research, RCIs were found to be useful when making diagnostic discriminations in older adults. PMID- 15797169 TI - Informed consent in clinical neuropsychology practice. Official statement of the National Academy of Neuropsychology. PMID- 15797170 TI - Current trends in the reimbursement of professional neuropsychological services. AB - Professional members of the National Academy of Neuropsychology (NAN) completed a survey focusing on clinicians' time requirements for activities, number of yearly claims to funding sources, average reimbursement rates, reimbursement rates associated with specific current procedural terminology (CPT) codes, managed care tolerance, hourly reimbursement rates, and other relevant practice issues such as incident-to. Clinicians continue to offer neuropsychological services through traditional assessment and evaluation activities, receive referrals from other healthcare professionals (i.e., neurologists, other medical specialists), and answer traditional referral questions related to patient diagnoses and determination of neuropsychological deficits. Two-thirds of respondents disapprove of managed care, and clinicians engage in activities which appear to minimize the ongoing changes in, and detrimental effects of, managed-care-based funding, in the form of dropping managed care panels, opting for fee-for-service, requiring self-pay patients, or spending more time in clinical activities for which there is a higher percentage of time approved such as forensic evaluations and evaluations for private practice. Clinicians continue to rely on managed care for their referrals even though they are only being reimbursed at about 50%. Overall, across service providers, clinicians are only receiving 60% of their hourly rate, and only 22% report receiving full compensation for their hourly rate. These results suggest that the effects of managed care have, for better or worse, altered the practice of clinical neuropsychology. PMID- 15797171 TI - Latent structure of the Wisconsin Card Sorting Test: a confirmatory factor analytic study. AB - The present study represents the first large scale confirmatory factor analysis of the Wisconsin Card Sorting Test (WCST). The results generally support the three factor solutions reported in the exploratory factor analysis literature. However, only the first factor, which reflects general executive functioning, is statistically sound. The secondary factors, while likely reflecting meaningful cognitive abilities, are less stable except when all subjects complete all 128 cards. It is likely that having two discontinuation rules for the WCST has contributed to the varied factor analytic solutions reported in the literature and early discontinuation may result in some loss of useful information. Continued multivariate research will be necessary to better clarify the processes underlying WCST performance and their relationships to one another. PMID- 15797172 TI - Detection and diagnosis of malingering in electrical injury. AB - This paper sought to demonstrate that diagnosable malingering does occur in Electrical Injury (EI) and examine the relationship of malingering to potential indicators of the presence and severity of neurological injury. Eleven consecutive EI patients seen for neuropsychological evaluation were presented. Over half the patients met the Slick et al. (1999) criteria for at least Probable MND. Most of the MND patients lacked evidence of a biologically meaningful exposure to electrical current. These findings highlight the importance of considering biological markers of neurological injury and of non-neurological factors, including effort/malingering, in the study of the neurocognitive consequences of EI. PMID- 15797173 TI - Test of Memory Malingering Performance is unaffected by laboratory-induced pain: implications for clinical use. AB - The Test of Memory Malingering (TOMM) is a well-validated and widely used forced choice symptom validity test. However, little is known about how TOMM performance is affected by pain. The present study evaluated the sensitivity of the TOMM to pain induced in healthy participants via the cold-presser test. Participants (n=20 per group) were administered the TOMM under one of three conditions: (1) standard instructions; (2) instructions to simulate pain-related memory deficit in pursuit of personal injury litigation; (3) while experiencing cold-induced pain. Results indicate that TOMM performance was unaffected by laboratory-induced moderate to severe pain and support the TOMM's use in evaluating clinical patients with pain. PMID- 15797175 TI - Automated neuropsychological assessment metrics (ANAM) measures of cognitive effects of Alzheimer's disease. AB - Eight individuals with Alzheimer's disease, and eight age-matched controls, were administered the MMSE, the Yesavage GDS, and a customized subset of the Automated Neuropsychological Assessment Metrics (ANAM) Battery. Accuracy (percent correct) and efficiency (number of correct responses per minute) of performance on six ANAM tasks were assessed. The patients' GDS scores indicated no depression. Although their MMSE scores (mean approximately 25) were significantly lower than those of the controls, they nonetheless indicated that the patients were still functioning at a fairly high level. Analysis of ANAM accuracy scores indicated that the patients were significantly impaired on three tasks measuring working memory. A discriminant function analysis revealed 93.8% correct classification. Analysis of ANAM efficiency scores revealed that except for simple reaction time, the patients were significantly impaired on all tasks. A discriminant function analysis correctly classified 100% of the participants. Given the small size of the groups in the present study, this finding especially underscores the sensitivity of ANAM to the cognitive effects of Alzheimer's disease, as indicated by the large effect sizes. The findings further indicate that ANAM might be capable of detecting more subtle effects of the disease at an earlier stage in its progress. PMID- 15797174 TI - Relationships between the WISC-III and the Cognitive Assessment System with Conners' rating scales and continuous performance tests. AB - The aim of this study was to examine the relationships between intelligence, ratings of behavior, and continuous performance test scores for a sample of 117 children aged 6-16 years who were referred to a specialty clinic. The sample was comprised of children who had a primary (45%) or secondary (36%) diagnosis of ADHD. All children were given the Wechsler Intelligence Scale for Children Third Edition (WISC-III), Cognitive Assessment System (CAS), Conners' Continuous Performance Test (CPT), and Conners' Parent and Teacher Rating Scales--Revised, Long Form. Correlations between Conners' Behavior Rating Scale and Conners' Continuous Performance Test were uniformly low and non-significant (the highest correlation was .17). Correlations between the WISC-III and Conners' Parent Rating Scale were all non-significant, but Teacher Ratings showed significant correlations between most of the WISC-III factors and the Cognitive Problems/Inattention scores. Few significant correlations were found between CPT with the WISC-III and CAS. These results suggest that practitioners should expect to find a lack of consistency between the scores provided by these measures and should be conservative of their use in clinical settings. PMID- 15797176 TI - A revision of A. L. Benton's Visual Retention Test (BVRT) in two parallel forms. AB - Selected items from various forms of the original Benton Visual Retention Test (BVRT) were used to construct two parallel forms with 20 items in each form. The extension of the test has improved its reliability, often criticized as being too low. By enhancing the reliability, we improved the sensitivity of the test for impairment, however the specificity remains unchanged in the revision. We examined the impact of age and education on the BVRT test scores. The modified forms were standardized for four age groups between 15 and 70 years. Based on the results of a Rasch analysis, we suggest that the administration of the revised BVRT may be discontinued after four consecutive failures without any serious effects on the test score. PMID- 15797177 TI - Neurofibromatosis 1: from lab bench to clinic. AB - Neurofibromatosis type 1 is a common autosomal dominant disorder in which affected children and adults develop both benign and malignant tumors. In addition to tumor formation, children with neurofibromatosis type 1 may exhibit specific learning disabilities, distinctive bony abnormalities, and hyperpigmented lesions (cafe-au-lait macules, skinfold freckling, and Lisch nodules). With the identification of the neurofibromatosis 1 gene in 1990, significant strides have been made towards elucidating the pathogenesis of specific clinical problems in neurofibromatosis type 1 and developing first generation, biologically based targeted therapies. Recent advances in mouse modeling have likewise yielded important insights into the genetic and cellular mechanisms underlying neurofibromatosis 1-associated tumor formation and learning disabilities. This review will focus on the clinical features of neurofibromatosis type 1, the molecular biology of the neurofibromatosis 1 gene, and the use of mouse modeling to recapitulate the human condition. PMID- 15797178 TI - Channelopathy: hypothesis of a common pathophysiologic mechanism in different forms of paroxysmal dyskinesia. AB - Paroxysmal dyskinesias are a rare heterogeneous group of neurologic disorders, characterized by transient sudden choreoathetoid or dystonic attacks without loss of consciousness. This study reports a family with six affected members in three generations, and two sporadic cases of paroxysmal dyskinesia. Familial cases of paroxysmal dyskinesia are affected by idiopathic long-lasting paroxysmal exertion induced dyskinesia and the sporadic cases by idiopathic short-lasting paroxysmal kinesigenic dyskinesia. Familial cases also suffer from epilepsy, mainly of generalized type, with benign outcome; one sporadic case is affected by migraine. Results presented in this neurophysiologic study include electromyography, somatosensory evoked potentials by median nerve stimulation, somatosensory evoked potentials by posterior tibial nerve stimulation, motor evoked potentials by magnetic transcranial cortical stimulation, visual evoked potentials, brainstem auditory evoked potentials, blink reflex, reflex H, and electroencephalography. The clinical and neurophysiologic findings presented here suggest a condition of hyperexcitability at the muscular and brain level, perhaps as a result of an ion channel disorder, which is in agreement with reports in the literature. PMID- 15797179 TI - Auditory steady-state responses in infants with perinatal brain injury. AB - Infants with perinatal brain injury present impairments in motor, visual, auditory, and cognitive functions. The most useful methods for detecting auditory alterations are auditory brainstem responses and otoacoustic emissions. Auditory steady-state responses have been reported as a reliable and objective technique for evaluating the hearing threshold. Auditory brainstem responses and auditory steady-state responses were carried out in 53 infants with perinatal brain injury and abnormal neurologic findings. With auditory brainstem responses, 33 (62.26%) infants presented normal and 20 abnormal results; 8 (15.09%) exhibited mild alterations, 8 (15.09%) moderate, and 4 (7.54%) severe alterations. With auditory steady-state responses, 17 (32.0%) infants were normal and 36 (67.9%) had abnormal results. When auditory steady-state responses were compared with auditory brainstem responses gold standard, the assessment gave 100% sensitivity, 51.51% specificity, 55.55% positive predictive value, and 100% negative predictive value. Abnormalities were mild in 21 (39.6%) infants, moderate in 10 (18.9%), and 5 (9.4%) exhibited severe hearing loss. We conclude that hearing loss is a frequent abnormality in infants with perinatal brain injury, and auditory steady-state responses have a high sensitivity for detecting hearing impairment, which is more evident in mild hearing loss for specific frequencies. PMID- 15797180 TI - Postneonatal epilepsy following amplitude-integrated EEG-detected neonatal seizures. AB - To assess the incidence of postneonatal epilepsy in term infants treated with antiepileptic drugs for neonatal seizure discharges that were detected with amplitude-integrated electroencephalography (aEEG), 206 term infants were monitored using this modality. They received antiepileptic drugs for clinical as well as subclinical neonatal seizures. Follow-up data were analyzed for the development of postneonatal epilepsy and for their neurodevelopmental outcome, assessed at 3, 9, 18 months, and 3 and 5 years of age. A total of 169 (82%) neonates received two or more antiepileptic drugs. Overall mortality was 39% (n = 80). Forty-one of the 126 survivors (33%) were abnormal at follow-up, and 12 of them developed postneonatal epilepsy (9.4%). Eighty-four children survived after hypoxic-ischemic encephalopathy grade II (n = 92), and 6 (7%) developed postneonatal epilepsy. In this subgroup, no postneonatal epilepsy was observed if seizures were controlled within 48 hours after birth and when not more than two antiepileptic drugs were required. Twenty-four children survived after an intracranial hemorrhage (n = 28), and only 1 (4%) developed postneonatal epilepsy. Eleven children survived after perinatal arterial stroke (n = 13), and 2 (18%) developed postneonatal epilepsy. In conclusion, the incidence of postneonatal epilepsy after treatment of clinical and subclinical neonatal seizures detected with continuous amplitude-integrated electroencephalography was 9.4%; This figure is lower than previously reported in children who only received treatment for clinical seizures. PMID- 15797181 TI - Resting EEG theta activity predicts cognitive performance in attention-deficit hyperactivity disorder. AB - Quantitative electroencephalography has contributed significantly to elucidating the neurobiologic mechanisms of attention-deficit hyperactivity disorder. The most consistent and robust electroencephalographic disturbance in attention deficit hyperactivity disorder has been abnormally increased theta band during resting conditions. Separate research using attention-demanding tests has elucidated cognitive disturbances that differentiate attention-deficit hyperactivity disorder. This study attempts to integrate electroencephalographic and neuropsychological indices to determine whether cognitive performance is specifically related to increased theta. Theta activity was recorded during a resting condition for 46 children/adolescents with attention-deficit hyperactivity disorder and their sex- and age-matched control subjects. Accuracy and reaction time during an auditory oddball and a visual continuous performance test were then recorded. Compared with control subjects, the attention-deficit hyperactivity disorder group manifested significantly increased (primarily left) frontal theta. Furthermore, the attention-deficit hyperactivity disorder group scored significantly delayed reaction time and decreased accuracy in both tasks. Correlation analysis revealed a significant relationship between frontal (primarily left) theta and oddball accuracy for the attention-deficit hyperactivity disorder group compared with a significant relationship between posterior (primarily right) theta and reaction time in the continuous performance test for the control group. These results indicate that spatial neurophysiologic deficits in attention-deficit hyperactivity disorder may be related to disturbances in signal detection. This observation has important implications for the role of trait-like biologic deficits in attention-deficit hyperactivity disorder predicting performance in information processing. PMID- 15797182 TI - Posterior resection for childhood epilepsy. AB - We reviewed our experience with posterior resection for intractable childhood epilepsy. Fifteen patients, seven males and eight females, were studied. Age of surgery was 18 months to 16 years. Nine patients had a parietal resection and six patients had an occipital resection. Surgical outcome was variable. Nine patients had an excellent outcome (Engel Class I or II), two patients had a poor outcome (Engel Class III), and four patients had a very poor outcome (Engel Class IV). Pathology at surgery included focal cortical dysplasias (4), brain tumors (4), tubers of tuberous sclerosis (2), cerebrovascular accidents (2), porencephalic cysts (1), and normal pathology (2). Complications included visual field loss in the occipital lobe patients (4/6). Posterior resection can be successful for children with intractable epilepsy originating in the parietal and occipital lobes. Invasive monitoring is necessary in some patients to establish the extent of the epileptic zone before surgery. A good surgical outcome is dependent on the presence of a circumscribed lesion on MRI and ability to surgically remove all the pathology. Visual abnormalities are an expected complication of surgery in the occipital lobe, but may improve over time. PMID- 15797183 TI - Outcomes at school age of preschool children with developmental language impairment. AB - In a prospective study, preschool children diagnosed with developmental language impairment were systematically reassessed during the early school years with standardized developmental (Battelle Developmental Inventory) and functional measures (Vineland Adaptive Behavior Scale). Of an original cohort of 70 children assessed and diagnosed at a mean age of 3.6 +/- 0.7 years, 43 were reassessed at a mean age of 7.4 +/- 0.7 years. Group performance on the Battelle overall was 71.9 +/- 8.2 with the lowest sub-domain score in communication at 69.5 +/- 8.9. On the Battelle, 67% of children fell below the 1.5 standard deviation (S.D.) cutoff signifying significant developmental concerns. Between 36% (gross motor) and 83% (communication) of the cohort performed at least 1.5 S.D. below the normative mean on the individual domains of the Battelle. Seventy-four percent were impaired in two or more domains of the Battelle. The group mean on the Vineland overall was 81.1 +/- 16.9 with between 19% (socialization) to 48% (communication) of the cohort scoring more than 1.5 S.D. below the mean on each of the sub-domains. Almost half of the cohort (20/42, 48%) manifested functional impairment in at least two domains of the Vineland. Univariate and multivariate analysis of potential predictor variables identified only female sex as being predictive of significantly poorer performance on the Vineland communication sub domain and the Vineland total score. Children with early developmental language impairment demonstrate persistent impairments in developmental and functional skills at school entry not limited to language. Deficits remain especially evident in the communication sub-domain. These results have implications with respect to later prognostication, family counseling, and devising a programmatic approach to this group of children. PMID- 15797184 TI - Mucopolysaccharidosis type II in females: case report and review of literature. AB - Mucopolysaccharidosis type II (Hunter disease, iduronate-2-sulfatase deficiency) was diagnosed in a 4-year-old female by demonstrating low iduronate-2-sulfatase activity both in leukocytes and fibroblasts and by the presence of a novel, complex rearrangement of the iduronate-2-sulfatase gene in heterozygous form. Mucopolysaccharidosis type II is inherited in an X-linked recessive manner and consequently females are rare. The disease phenotype in this case is due to complete unilateral inactivation of the nonmutant paternal X chromosome of the patient. The case presented here underscores the fact that a diagnosis of mucopolysaccharidosis type II should be suspected in any female who presents with the relevant clinical symptoms. PMID- 15797185 TI - Episodic somnolence in an infant with Riley-Day syndrome. AB - Familial dysautonomia is an autosomal recessive congenital neuropathy that occurs almost exclusively in the Ashkenazi Jewish population and has rarely been diagnosed in the neonatal period in unaffected families. This report describes a patient who, during the neonatal period, had episodes of marked decrease in the level of consciousness with durations of 4-15 hours. Other signs and symptoms included the absence of fungiform papillae of the tongue, areflexia, and failure to thrive. The diagnosis was confirmed by the demonstration of mutations in the IkappaB kinase complex-associated protein gene with the identification of IVS20 (+6T --> C) which is responsible for more than 99.5% of known Ashkenazi Jewish patients with familial dysautonomia. The prognosis of this disease and the possibility of genetic counseling are clearly related with an early definitive diagnosis, and this patient illustrates the importance of episodes of somnolence as a possible sign of familial dysautonomia. PMID- 15797186 TI - Arterial spasm following perimesencephalic nonaneurysmal subarachnoid hemorrhage in a pediatric patient. AB - Perimesencephalic nonaneurysmal hemorrhage is a benign form of subarachnoid hemorrhages. This entity is well recognized as a distinct type of subarachnoid hemorrhage in adults. However, perimesencephalic nonaneurysmal subarachnoid hemorrhage in pediatric patients is not well recognized. Angiographic changes such as vasospasm are uncommon in patients, especially in pediatric patients suffering from this type of hemorrhage. This case study reports a 12-year-old male who suffered from perimesencephalic nonaneurysmal subarachnoid hemorrhage. Cerebral carotid angiography performed on the tenth day of the posthemorrhagic period revealed severe vasospasm affecting the basilar artery. The patient, treated symptomatically, was discharged after improvement. One year later, magnetic resonance angiography revealed completely normal features. PMID- 15797187 TI - Fatal neurologic involvement in pediatric Wegener's granulomatosis. AB - Wegener's granulomatosis is a potentially life-threatening vasculitis with widely variable presentation. Only three pediatric cases with severe central nervous system involvement are reported in the literature. Early fatal outcome as described here is exceptional. This report describes a 13-year-old female with typical skin lesions, proteinuria, and renal failure initially misdiagnosed as a Schoenlein-Henoch purpura. A kidney biopsy revealed severe extracapillary proliferation in 70% of the analyzed glomeruli but no granuloma. In spite of methylprednisolone pulse therapy and oral high-dose prednisone, end-stage renal failure was reached 4 months later, necessitating peritoneal dialysis. Three months later she presented with pulmonary hemorrhage and positive antineutrophil cytoplasmic antibodies suggesting Wegener's granulomatosis. This episode was controlled by methylprednisolone pulses. Seven months later she presented generalized seizures and coma, suggesting central nervous system involvement confirmed by magnetic resonance imaging. Methylprednisolone pulses and intravenous immunoglobulins led to neurologic improvement. Oral methotrexate was then introduced for long-term disease control. Another severe relapse of central nervous system vasculitis did not respond to any applied therapies and led to death 16 months after initial symptoms. This case emphasizes the need for activity scores to identify patients at risk for progressive systemic vasculitis requiring early and long-term aggressive immunosuppressive therapy. PMID- 15797188 TI - Superior sagittal sinus thrombosis in steroid-resistant nephrotic syndrome. AB - An 8(1/2)-year-old-female child with steroid-resistant nephrotic syndrome developed sagittal sinus thrombosis while on pulse therapy with corticosteroids, presenting with recurrent vomiting, headache, and impaired consciousness. The diagnosis was established by cranial computed tomography, magnetic resonance imaging, and magnetic resonance angiography. She gradually recovered without neurologic sequelae while being treated with low-molecular-weight heparin (2 mg/kg/day). Sagittal sinus thrombosis consists of a rare and probably underdiagnosed complication of childhood nephrotic syndrome. PMID- 15797189 TI - Poland-Mobius syndrome and cocaine abuse: a relook at vascular etiology. AB - This report presents a severe case of Poland-Mobius syndrome with central apnea at birth, ventilator-dependent, and with brainstem calcifications. The newborn had unilateral defect of the right pectoralis muscle, breast, and limb. He manifested bilateral paralysis of the cranial nerves resulting in shallow respiration, apnea, and dysphagia. He finally required tracheostomy and gastrostomy. Maternal history revealed multiple uses of cocaine during the first 3 months of pregnancy. The patient supports the Poland-Mobius combination and the possibility of vascular disruption sequence. PMID- 15797190 TI - A new mutation of IGHMBP2 gene in spinal muscular atrophy with respiratory distress type 1. AB - This report presents a new mutation in the first Japanese female infant with spinal muscular atrophy with respiratory distress type 1. She manifested the characteristic clinical features, including early-onset respiratory failure due to diaphragmatic paralysis and severe distal muscle weakness. Muscle biopsy in the femoral muscle indicated massive neurogenic changes. Sural nerve biopsy disclosed a moderate reduction of myelinated fibers, predominantly reduced large fibers. She had a novel homozygous missense mutation 2685 C -->A, leading to a T879K substitution in the immunoglobulin mu-binding protein 2 gene. Both parents were heterozygous for this mutation. PMID- 15797192 TI - Febrile and afebrile or provoked and unprovoked seizures? PMID- 15797194 TI - Transcriptional regulation by the numbers: models. AB - The expression of genes is regularly characterized with respect to how much, how fast, when and where. Such quantitative data demands quantitative models. Thermodynamic models are based on the assumption that the level of gene expression is proportional to the equilibrium probability that RNA polymerase (RNAP) is bound to the promoter of interest. Statistical mechanics provides a framework for computing these probabilities. Within this framework, interactions of activators, repressors, helper molecules and RNAP are described by a single function, the "regulation factor". This analysis culminates in an expression for the probability of RNA polymerase binding at the promoter of interest as a function of the number of regulatory proteins in the cell. PMID- 15797195 TI - Transcriptional regulation by the numbers: applications. AB - With the increasing amount of experimental data on gene expression and regulation, there is a growing need for quantitative models to describe the data and relate them to their respective context. Thermodynamic models provide a useful framework for the quantitative analysis of bacterial transcription regulation. This framework can facilitate the quantification of vastly different forms of gene expression from several well-characterized bacterial promoters that are regulated by one or two species of transcription factors; it is useful because it requires only a few parameters. As such, it provides a compact description useful for higher-level studies (e.g. of genetic networks) without the need to invoke the biochemical details of every component. Moreover, it can be used to generate hypotheses on the likely mechanisms of transcriptional control. PMID- 15797196 TI - DNA looping in gene regulation: from the assembly of macromolecular complexes to the control of transcriptional noise. AB - The formation of DNA loops by the binding of proteins and protein complexes at distal DNA sites plays a central role in many cellular processes, such as transcription, recombination and replication. Important thermodynamic concepts underlie the assembly of macromolecular complexes on looped DNA. The effects that this process has on the properties of gene regulation extend beyond the traditional view of DNA looping as a mechanism to increase the affinity of regulatory molecules for their cognate sites. Recent developments indicate that DNA looping can also lead to the suppression of cell-to-cell variability, the control of transcriptional noise, and the activation of cooperative interactions on demand. PMID- 15797197 TI - Revisited gene regulation in bacteriophage lambda. AB - The contribution of bacteriophage lambda to gene control research is far from over. A revised model of the lambda genetic switch includes extra cooperativity through octamerization of the cI repressor protein, mediated by long-range DNA looping. Structural analysis reveals remarkably subtle transcriptional activation by cI. The action of cI, activation by cII, and aspects of antitermination by N and Q all confirm the utility and versatility of simple, weak adhesive interactions mediated by nucleic acid tethers. New genetic and quantitative analysis of the lambda gene network is challenging cherished ideas about how complex behaviours emerge from this regulatory system. PMID- 15797198 TI - The structure and function of the bacterial chromosome. AB - Advances in microscopic and cell biological techniques have considerably improved our understanding of bacterial chromosome organization and dynamics. The nucleoid was formerly perceived to be an amorphous entity divided into ill-defined domains of supercoiling that are randomly deposited in the cell. Recent work, however, has demonstrated a remarkable degree of spatial organization. A highly ordered chromosome structure, established while DNA replication and partitioning are in progress, is maintained and propagated during growth. Duplication of the chromosome and partitioning of the newly generated daughter strands are interwoven processes driven by the dynamic interplay between the synthesis, segregation and condensation of DNA. These events are intimately coupled with the bacterial cell cycle and exhibit a previously unanticipated complexity reminiscent of eukaryotic systems. PMID- 15797199 TI - The key to development: interpreting the histone code? AB - Developmental stages in multicellular organisms proceed according to a temporally and spatially precise pattern of gene expression. It has become evident that changes within the chromatin structure brought about by covalent modifications of histones are of crucial importance in determining many biological processes, including development. Numerous studies have provided evidence that the enzymes responsible for the modifications of histones function in a coordinated pattern to control gene expression in the short term and, through the transferral of these modifications by inheritance to their progeny, in the long term. PMID- 15797200 TI - Centromeric chromatin: what makes it unique? AB - Centromeres represent the final frontier of eukaryotic genomes. Although they are defining features of chromosomes--the points at which spindle microtubules attach -the fundamental features that distinguish them from other parts of the chromosome remain mysterious. The function of centromeres is conserved throughout eukaryotic biology, but their DNA sequences are not. Rather, accumulating evidence favors chromatin-based centromeric identification. To understand how centromeric identity is maintained, researchers have studied DNA-protein interactions at native centromeres and ectopic "neocentromeres". Other studies have taken a comparative approach focusing on centromere-specific proteins, of which mammalian CENP-A and CENP-C are the prototypes. Elucidating the assembly and structure of chromatin at centromeres remain key challenges. PMID- 15797201 TI - Chromatin remodeling complexes: strength in diversity, precision through specialization. AB - Chromatin remodeling complexes all have an ATPase subunit important for nucleosome remodeling, but are diversified and specialized by additional associated proteins. Remodelers have well-established roles in a wide range of chromosomal processes, including transcriptional regulation and chromatin assembly. Recent work, however, has revealed remarkable new functions for remodelers, such as histone variant deposition, cohesin function, and RNA transcript elongation and termination. Remodeler complexes are tailored both compositionally and mechanistically to perform particular chromatin functions. PMID- 15797202 TI - Controlling DNA methylation: many roads to one modification. AB - Genetic, biochemical and cytological studies on DNA methylation in several eukaryotic organisms have resulted in leaps of understanding in the past three years. Discoveries of mechanistic links between DNA methylation and histone methylation, and between these processes and RNA interference (RNAi) machineries have reinvigorated the field. The details of the connections between DNA methylation, histone modifications and RNA silencing remain to be elucidated, but it is already clear that no single pathway accounts for all DNA methylation found in eukaryotes. Rather, different taxa use one or more of several general mechanisms to control methylation. Despite recent progress, classic questions remain, including: What are the signals for DNA methylation? Are "de novo" and "maintenance" methylation truly separate processes? How is DNA methylation regulated? PMID- 15797203 TI - MicroRNAs: a developing story. AB - Considering the intense genetic efforts applied to understanding development, it is surprising that a relatively large class of regulatory genes has newly surfaced. The first microRNA gene and its developmental role were described more than ten years ago, but only recently have we fully appreciated the broad and abundant presence of such genes. MicroRNAs are approximately 22 nucleotide RNAs that use antisense complementarity to inhibit expression of specific mRNAs. Recent studies of restricted expression patterns and functional roles have implicated specific microRNAs in complex genetic pathways regulating embryogenesis, hematopoiesis, neuronal differentiation and Hox-mediated development. PMID- 15797204 TI - Recognition and modification of seX chromosomes. AB - Flies, worms and mammals employ dosage compensation complexes that alter chromatin or chromosome structure to equalize X-linked gene expression between the sexes. Recent work has improved our understanding of how dosage compensation complexes achieve X chromosome-wide association and has provided significant insight into the epigenetic modifications directed by these complexes to modulate gene expression. In flies, the prevailing view that dosage compensation complexes assemble on the X chromosome at approximately 35 chromatin-entry sites and then spread in cis to cover the chromosome has been re-evaluated in light of the evidence that these chromatin-entry sites are not required for localization of the complex. By contrast, identification of discrete recruitment elements indicates that nucleation at and spread from a limited number of sites directs dosage compensation complex localization on the worm X-chromosome. Studies in flies and mammals have extended our understanding of how ribonucleoprotein complexes are used to modify X chromatin, for either activation or repression of transcription. Finally, evidence from mammals suggests that the chromatin modifications that mediate dosage compensation are very dynamic, because they are established, reversed and re-established early in development. PMID- 15797205 TI - Computational methods for transcriptional regulation. AB - How is the information from a thousand gene-expression arrays, the location of more than two hundred regulatory factors, and nine sequenced genomes to be integrated into a global view of the regulatory network in budding yeast? Computational methods that fit incomplete noisy data provide the outlines of regulatory pathways, but the errors are not quantified. In the fly, embryonic patterning has proved amenable to computational prediction, but only when the DNA binding preferences of the relevant factors are taken into account. In both these model organisms, simply restricting attention to regulatory sequences that align with related species (i.e. "conserved") discards much information regarding what is functional. PMID- 15797206 TI - Inhibitex, Inc: antibody anti-infectives. PMID- 15797207 TI - Exploiting Pseudomonas putida for drug development. AB - In this issue of Chemistry & Biology, a strategy that combines large DNA fragment recombineering in Escherichia coli and heterologous expression in Pseudomonas putida is described. The work focuses on myxochromide S, a natural compound produced by Stigmatella aurantiaca. PMID- 15797208 TI - Adding specificity to artificial transcription activators. AB - In this issue, Mapp and colleagues describe a significant advance in the design of artificial transcription activators that function in a cell-type-specific manner. [1] The authors show that peptides selected for binding a component of the yeast transcription complex require its presence for effective transcriptional activation. PMID- 15797209 TI - Scratch n' screen for inhibitors of cell migration. AB - Yarrow et al. have identified a small molecule inhibitor of cell migration, 3-(4 pyridyl)indole ("Rockout"), that targets Rho-kinase via a novel screening method using a scratch wound healing assay adapted to a high-throughput format and automated microscopy. PMID- 15797210 TI - Structural insights into biological roles of protein-glycosaminoglycan interactions. AB - The extracellular environment is largely comprised of complex polysaccharides, which were historically considered inert materials that hydrated the cells and contributed to the structural scaffolds. Recent advances in development of sophisticated analytical techniques have brought about a dramatic transformation in understanding the numerous biological roles of these complex polysaccharides. Glycosaminoglycans (GAGs) are a class of these polysaccharides, which bind to a wide variety of proteins and signaling molecules in the cellular environment and modulate their activity, thus impinging on fundamental biological processes. Despite the importance of GAGs modulating biological functions, there are relatively few examples that demonstrate specificity of GAG-protein interactions, which in turn define the structure-function relationships of these polysaccharides. Focusing on heparin/heparan (HSGAGs) and chondroitin/dermatan sulfate (CSGAGs), this review provides structural insights into the oligosaccharide-protein interactions and discusses some key and challenging aspects of understanding GAG structure-function relationships. PMID- 15797211 TI - Conversion of the monomeric red fluorescent protein into a photoactivatable probe. AB - Photoactivatable fluorescent proteins bring new dimension to the analysis of protein dynamics in the cell. Protein tagged with a photoactivatable label can be visualized and tracked in a spatially and temporally defined manner. Here, we describe a basic rational design strategy to develop monomeric photoactivatable proteins using site-specific mutagenesis of common monomeric red-shifted fluorescent proteins. This strategy was applied to mRFP1, which was converted into probes that are photoactivated by either green or violet light. The latter photoactivatable variants, named PA-mRFP1s, exhibited a 70-fold increase of fluorescence intensity resulting from the photoconversion of a violet-light absorbing precursor. Detailed characterization of PA-mRFP1s was performed with the purified proteins and the proteins expressed in mammalian cells where the photoactivatable properties were preserved. PA-mRFP1s were used as protein tags to study the intracellular dynamics of GTPase Rab5. PMID- 15797212 TI - Structural basis of swinholide A binding to actin. AB - Marine toxins targeting the actin cytoskeleton represent a new and promising class of anti-cancer compounds. Here we present a 2.0 A resolution structure of swinholide A, a marine macrolide, bound to two actin molecules. The structure demonstrates that the actin dimer in the complex does not represent a physiologically relevant entity, for the two actin molecules do not interact with each other. The swinholide A actin binding site is the same as that targeted by toxins of the trisoxazole family and numerous actin binding proteins, highlighting the importance of this site in actin polymerization. The observed structure reveals the mechanism of action of swinholide A and provides a structural framework about which to design new agents directed at the cytoskeleton. PMID- 15797213 TI - The neocarzinostatin biosynthetic gene cluster from Streptomyces carzinostaticus ATCC 15944 involving two iterative type I polyketide synthases. AB - The biosynthetic gene cluster for the enediyne antitumor antibiotic neocarzinostatin (NCS) was localized to 130 kb continuous DNA from Streptomyces carzinostaticus ATCC15944 and confirmed by gene inactivation. DNA sequence analysis of 92 kb of the cloned region revealed 68 open reading frames (ORFs), 47 of which were determined to constitute the NCS cluster. Sequence analysis of the genes within the NCS cluster suggested dNDP-D-mannose as a precursor for the deoxy aminosugar, revealed two distinct type I polyketide synthases (PKSs), and supported a convergent model for NCS chromophore biosynthesis from the deoxy aminosugar, naphthoic acid, and enediyne core building blocks. These findings shed light into deoxysugar biosynthesis, further support the iterative type I PKS paradigm for enediyne core biosynthesis, and unveil a mechanism for microbial polycyclic aromatic polyketide biosynthesis by an iterative type I PKS. PMID- 15797214 TI - The adamantane-derived bananins are potent inhibitors of the helicase activities and replication of SARS coronavirus. AB - Bananins are a class of antiviral compounds with a unique structural signature incorporating a trioxa-adamantane moiety covalently bound to a pyridoxal derivative. Six members of this class of compounds: bananin, iodobananin, vanillinbananin, ansabananin, eubananin, and adeninobananin were synthesized and tested as inhibitors of the SARS Coronavirus (SCV) helicase. Bananin, iodobananin, vanillinbananin, and eubananin were effective inhibitors of the ATPase activity of the SCV helicase with IC50 values in the range 0.5-3 microM. A similar trend, though at slightly higher inhibitor concentrations, was observed for inhibition of the helicase activities, using a FRET-based fluorescent assay. In a cell culture system of SCV, bananin exhibited an EC50 of less than 10 microM and a CC50 of over 300 microM. Kinetics of inhibition are consistent with bananin inhibiting an intracellular process or processes involved in SCV replication. PMID- 15797215 TI - Functional specificity of artificial transcriptional activators. AB - Misregulated transcription is linked to many human diseases, and thus artificial transcriptional activators are highly desirable as mechanistic tools and as replacements for their malfunctioning natural counterparts. We previously reported two artificial transcriptional activation domains obtained from synthetic peptide libraries screened for binding to the yeast transcription protein Med15(Gal11). Here we demonstrate that the transcriptional potency of the Med15 ligands is increased through straightforward structural alterations. These artificial activation domains upregulate transcription via specific Med15 binding interactions and do not function in mammalian cells, which lack Med15. This functional specificity stands in contrast to most natural or artificial activation domains that function across all eukaryotic cell types. The results indicate that the screening strategy holds excellent promise for identifying peptide and small molecule transcriptional activators that function by unique mechanisms with advantageous specificity properties. PMID- 15797216 TI - Gold nanoparticles as a colorimetric sensor for protein conformational changes. AB - Spherical gold nanoparticles and flat gold films are prepared in which yeast iso 1-cytochrome c (Cyt c) is covalently bound to the gold surface by a thiol group in the cystein 102 residue. Upon exposure to solutions of different pH, bound Cyt c unfolds at low pH and refolds at high pH. This conformational change causes measurable shifts in the color of the coated nanoparticle solutions detected by UV-VIS absorption spectroscopy and in the refractive index (RI) of the flat gold films detected by surface plasmon resonance (SPR) spectroscopy. Both experiments demonstrate the same trend with pH, suggesting the use of protein-covered gold nanoparticles as a simple colorimetric sensor for conformational change. PMID- 15797217 TI - Identification of amino acids that promote specific and rigid TAR RNA-tat protein complex formation. AB - The Tat protein and the transactivation responsive (TAR) RNA form an essential complex in the HIV lifecycle, and mutations in the basic region of the Tat protein alter this RNA-protein molecular recognition. Here, EPR spectroscopy was used to identify amino acids, flanking an essential arginine of the Tat protein, which contribute to specific and rigid TAR-Tat complex formation by monitoring changes in the mobility of nitroxide spin-labeled TAR RNA nucleotides upon binding. Arginine to lysine N-terminal mutations did not affect TAR RNA interfacial dynamics. In contrast, C-terminal point mutations, R56 in particular, affected the mobility of nucleotides U23 and U38, which are involved in a base triple interaction in the complex. This report highlights the role of dynamics in specific molecular complex formation and demonstrates the ability of EPR spectroscopy to study interfacial dynamics of macromolecular complexes. PMID- 15797218 TI - Use of the tubulin bound paclitaxel conformation for structure-based rational drug design. AB - A new computational docking protocol has been developed and used in combination with conformational information inferred from REDOR-NMR experiments on microtubule bound 2-(p-fluorobenzoyl)paclitaxel to delineate a unique tubulin binding structure of paclitaxel. A conformationally constrained macrocyclic taxoid bearing a linker between the C-14 and C-3'N positions has been designed and synthesized to enforce this "REDOR-taxol" conformation. The novel taxoid SB-T 2053 inhibits the growth of MCF-7 and LCC-6 human breast cancer cells (wild-type and drug resistant) on the same order of magnitude as paclitaxel. Moreover, SB-T 2053 induces in vitro tubulin polymerization at least as well as paclitaxel, which directly validates our drug design process. These results open a new avenue for drug design of next generation taxoids and other microtubule-stabilizing agents based on the refined structural information of drug-tubulin complexes, in accordance with typical enzyme-inhibitor medicinal chemistry precepts. PMID- 15797219 TI - Heterologous expression of a myxobacterial natural products assembly line in pseudomonads via red/ET recombineering. AB - Natural products of microbial origin are widely used as pharmaceuticals and in agrochemistry. These compounds are often biosynthesized by multifunctional megasynthetases whose genetic engineering and heterologous expression offer considerable promise, especially if the natural hosts are genetically difficult to handle, slow growing, unculturable, or even unknown. We describe a straightforward strategy that combines the power of advanced DNA engineering (recombiogenic cloning) in Escherichia coli with the utility of pseudomonads as the heterologous host for the analysis and mutagenesis of known and unknown secondary metabolite pathways. The myxochromide S biosynthetic gene cluster from Stigmatella aurantiaca was rebuilt and engineered in E. coli to contain the elements required for expression in pseudomonads. The successful production in Pseudomonas putida, at unprecedented levels, demonstrates the feasibility of the new approach to the analysis and mutagenesis of these important pathways. PMID- 15797220 TI - Delineation of the chemical pathways underlying nitric oxide-induced homologous recombination in mammalian cells. AB - Inflammation is an important risk factor for cancer. During inflammation, macrophages secrete nitric oxide (NO*), which reacts with superoxide or oxygen to create ONOO- or N2O3, respectively. Although homologous recombination causes DNA sequence rearrangements that promote cancer, little was known about the ability of ONOO- and N2O3 to induce recombination in mammalian cells. Here, we show that ONOO- is a potent inducer of homologous recombination at an integrated direct repeat substrate, whereas N2O3 is relatively weakly recombinogenic. Furthermore, on a per lesion basis, ONOO(-)-induced oxidative base lesions and single-strand breaks are significantly more recombinogenic than N2O3-induced base deamination products, which did not induce detectable recombination between plasmids. Similar results were observed in mammalian cells from two different species. These results suggest that ONOO(-)-induced recombination may be an important mechanism underlying inflammation-induced cancer. PMID- 15797221 TI - Synthetic erythropoietic proteins: tuning biological performance by site-specific polymer attachment. AB - Chemical synthesis in combination with precision polymer modification allows the systematic exploration of the effect of protein properties, such as charge and hydrodynamic radius, on potency using defined, homogeneous conjugates. A series of polymer-modified synthetic erythropoiesis proteins were constructed that had a polypeptide chain similar to the amino acid sequence of human erythropoietin but differed significantly in the number and type of attached polymers. The analogs differed in charge from +5 to -26 at neutral pH and varied in molecular weight from 30 to 54 kDa. All were active in an in vitro cell proliferation assay. However, in vivo potency was found to be strongly dependent on overall charge and size. The trends observed in this study may serve as starting points for the construction of more potent synthetic EPO analogs in the future. PMID- 15797222 TI - Screening for cell migration inhibitors via automated microscopy reveals a Rho kinase inhibitor. AB - Small-molecule kinase inhibitors are predominantly discovered in pure protein assays. We have discovered an inhibitor of Rho-kinase (ROCK) through an image based, high-throughput screen of cell monolayer wound healing. Using automated microscopy, we screened a library of approximately 16,000 compounds finding many that affected cell migration or cell morphology as well as compounds that blocked mitotic progression. We tested approximately 200 compounds in a series of subassays and chose one, 3-(4-pyridyl)indole (Rockout), for more detailed characterization. Rockout inhibits blebbing and causes dissolution of actin stress fibers, phenocopying Rho-kinase inhibitors. Testing Rho-kinase activity in vitro, Rockout inhibits with an IC50 of 25 microM ( approximately 5-fold less potent than Y-27632) but has a similar specificity profile. We also profile the wound healing assay with a library of compounds with known bioactivities, revealing multiple pathways involved in the biology. PMID- 15797223 TI - Speciation and biosynthetic variation in four dictyoceratid sponges and their cyanobacterial symbiont, Oscillatoria spongeliae. AB - Four species of marine sponges (Phylum Porifera, Order Dictyoceratida), which contain the filamentous cyanobacterial symbiont Oscillatoria spongeliae, were collected from four locations in Palau. The halogenated natural products associated with the symbiont were characterized from each sample, revealing that each species contained either chlorinated peptides, brominated diphenyl ethers, or no halogenated compounds. Analysis of the host sponges and the symbionts indicated that each species of sponge contained a distinct strain of morphologically similar cyanobacteria. Although cospeciation may be present in this group, we have identified that at least one host switching event has occurred in this symbiosis. Only the strain of O. spongeliae in the sponge containing the chlorinated compounds possessed genes involved in the biosynthesis of chlorinated leucine precursors, indicating that the chemical variation observed in these animals has a genetic foundation. PMID- 15797224 TI - Rectal complications after modern radiation for prostate cancer: a colorectal surgical challenge. AB - The operative management of rectal complications after radiation for prostate cancer has been incompletely studied. Our aim was to determine a logical surgical approach to these severe rectal complications. From an institutional database, we identified 5719 patients who were evaluated between 1990 and 2003 with a history of prostate cancer that was treated with radiation. Fourteen patients were identified from this group who underwent operative intervention for complications stemming from radiation. Charts were retrospectively reviewed for demographics, prostate cancer treatment, rectal symptoms, diagnostic techniques, operative interventions, and outcome. Ten patients (71%) had documented rectourethral fistulas. An additional four patients (29%) had either transfusion-dependent rectal bleeding or intractable fecal incontinence. Using a surgical algorithm, we proceeded with fecal diversion alone (20%), urinary and fecal diversion alone (50%), and primary repair with or without a tissue flap and fecal diversion (29%) in the 14 affected patients. Symptomatic improvement and resolution of these three complications occurred in 12 (85%) of patients. However, only 2 (15%) were able to retain their intestinal continuity to achieve this outcome. The introduction of a step-wise approach to this problem has resulted in symptomatic resolution in the majority of patients. However, this is achieved at the cost of permanent fecal and sometimes urinary diversion. PMID- 15797225 TI - Fas/FasL play a central role in pancreatitis-induced hepatocyte apoptosis. AB - Liver injury is a clinical prognostic indicator in acute pancreatitis (AP). We have demonstrated that Kupffer cell-derived FasL mediates liver injury during AP and sought to determine its role in AP-induced hepatocyte apoptosis. AP was induced in National Institutes of Health (NIH) Swiss mice, C57/C57, and Fas-/-, FasL-/- mice by a choline-deficient ethionine-supplement diet. Liver Fas, FasL, p38-mitogen activated phosphokinase (p38-MAPK), poly-ADP ribose polymerase (PARP), and cytochrome C were measured by immunoblotting. Apoptosis was assessed by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) and DNA fragmentation (ELISA). AP upregulated liver FasL (4280 +/- 580 vs. 733 +/ 336), Fas (2866 +/- 595 vs. 649 +/- 111), cytochrome C (6980 +/- 237 vs. 903 +/- 156), and PARP (6393 +/- 591 vs. 466 +/- 261) as well as increased TUNEL staining (40 +/- 2 vs. 14 +/- 1) and DNA fragmentation (all P < 0.03 vs. control). In FasL /- and Fas-/- mice, AP-induced upregulation of p38-MAPK, PARP, and cytochrome C was significantly attenuated (all P < 0.01 compared to C57/C57 control). In addition, AP-induced DNA fragmentation was reduced 60% in Fas-/- and FasL-/- mice (P < 0.01 vs. C57/C57). AP induces apoptosis by transcriptional activation of Fas/FasL. AP-induced apoptosis was significantly reduced in Fas and FasL knockout mice along with downregulation of p38-MAPK, PARP, and cytochrome C, thereby suggesting a central role for Fas/FasL in hepatocyte apoptosis. The manipulation of interactions between Kupffer cell-derived FasL and hepatocytes may have important therapeutic implications. PMID- 15797226 TI - The utility of laparoscopic assessment in the preoperative staging of suspected hilar cholangiocarcinoma. AB - The aim of this study was to review the role of laparoscopic assessment in the staging algorithm of suspected hilar cholangiocarcinoma and to identify factors highly likely to be associated with unresectable disease. Data prospectively collected between 1992 and 2003 were analyzed. Demographics, symptoms, preoperative radiologic staging, laparoscopic assessment, and final outcome were recorded. Yield was defined as the number of unresectable patients detected by laparoscopic assessment divided by the total number of patients undergoing laparoscopic assessment. Accuracy was defined as the number of unresectable patients detected by laparoscopic assessment divided by the total number of unresectable cases. Eighty-four patients underwent laparoscopic assessment for suspected hilar cholangiocarcinoma, of which 20 (23.8%) underwent resection. The yield from laparoscopy alone was 24.3% (20 of 82), which increased to 41.5% (35 of 82) with the addition of intraoperative ultrasound. The overall accuracy was 53.1% (35 of 66). The use of a preoperative radiologic staging system predicted the likelihood of unresectable disease (P=0.007). The use of laparoscopic assessment in the preoperative staging of patients with suspected hilar cholangiocarcinoma is justified given it will spare 42.2% of patients an unnecessary laparotomy. Accurate staging of cholangiocarcinoma remains a challenge, but the use of a preoperative radiologic staging system may help to stratify a patient's risk of unresectable disease. PMID- 15797228 TI - Selective portal vein clamping for radiofrequency ablation of hepatocellular carcinoma with portal vein invasion. AB - Surgical resection provides potential cure for patients with hepatocellular carcinoma. Unfortunately, resection is suitable in only about 10-37% of patients because of the limited hepatic functional reserve from the underlying chronic liver disease in the majority of patients. Survival of patients with unresectable diseases, especially those with portal vein tumor invasion, remains very poor. Radiofrequency ablation (RFA) is a form of locoregional therapy that allows a selected group of previously inoperable patients to be treated. However, problems with RFA leading to induced portal vein thrombosis have been reported in the literature. Nevertheless, patients with portal vein tumor invasion may be considered for radiofrequency tumor ablation to improve survival. We report the case of a patient with hepatocellular carcinoma with left portal vein invasion. Complete tumor ablation was achieved after RFA with left portal vein clamping. He remained disease free both radiologically and biochemically 6 months after the operation. PMID- 15797227 TI - The role of ciprofloxacin in prolonging polyethylene biliary stent patency: a multicenter, double-blinded effectiveness study. AB - Plastic stents are the mainstay of the palliation of malignant jaundice but are complicated by recurrent obstruction. Previous trials have failed to demonstrate any improvement in patency with the use of antibiotics. Patients with malignant jaundice were randomized in a double-blind fashion, after polyethylene stent insertion, to receive ciprofloxacin or placebo. After successful stent decompression, there were 50 patients in the treatment arm and 44 in the placebo. There were 14 (33%) episodes of stent occlusion in the ciprofloxacin group versus 23 (49%) in placebo (chi(2) test, P=0.115). There was no significant difference in patency (log-rank test, P=0.17). There were significantly fewer episodes of cholangitis with ciprofloxacin: 10 (23%) versus 21 (42%) in the placebo (P=0.047). The ciprofloxacin group also demonstrated a significant improvement in the Social Function domain of the SF-36 Quality of Life Survey at 1 month (paired T test, P=0.03). The other domains of the SF-36 were not different, nor was survival (log rank, P=0.80). There is insufficient evidence to show that prophylactic ciprofloxacin can prolong plastic biliary stent patency. The observed trends suggest that ciprofloxacin significantly decreases the incidence of cholangitis and results in improvements in certain aspects of quality of life. PMID- 15797229 TI - Xanthogranulomatous cholecystitis in laparoscopic surgery. AB - Xanthogranulomatous cholecystitis (XGC) is one presentation of cholecystitis and can be a cause of difficulty in cholecystectomy. We reviewed the clinical files of 12,426 patients who had undergone cholecystectomy. In this group, there were 182 cases of XGC, and 41 of these patients had undergone laparoscopic surgery. Patients with XGC represented 1.46% of the cholecystectomies that were performed. Of the 41 patients who underwent laparoscopic surgery, 27 were men (66%) and 14 were women (34%) (average age, 52 years). A total of 36 patients (88%) presented with a chronic condition. XGC was found to be associated with lithiasis in 85%, with jaundice in 22%, and with cancer in 2.4% (one patient). A total of 33 patients (80%) required conversion to open surgery, because of technical difficulties; of these patients, 64% underwent partial cholecystectomy. We conclude that XGC creates difficulty at laparoscopy and therefore any preoperative suspicion of XGC should cause the clinician to consider open cholecystectomy. PMID- 15797230 TI - Gastric cancer developing in the stomach after pylorus-preserving pancreaticoduodenectomy with pancreaticogastrostomy: case report and review of the literature. AB - A 59-year-old woman underwent surgery for uterine corpus cancer in March 1998. She also underwent pylorus-preserving pancreaticoduodenectomy with pancreaticogastrostomy for common bile duct cancer in November 1998. She was followed up at our outpatient clinic after pylorus-preserving pancreaticoduodenectomy. In November 2002, her carcinoembryonic antigen level became elevated and abdominal ultrasound revealed a huge tumor. Gastroscopy showed a Borrmann type 3 tumor at the anastomosis of the pancreaticogastrostomy, and a biopsy revealed adenocarcinoma. With a diagnosis of advanced gastric cancer, she underwent total gastrectomy, splenectomy, and residual pancreatectomy in January 2003. The pathologic findings revealed that the gastric cancer was separated from the pancreas, suggesting that the cancer had developed from the stomach. The present report describes a rare case of gastric cancer that had developed at the anastomosis of a pancreaticogastrostomy. PMID- 15797231 TI - Resection of the celiac axis for invasive pancreatic cancer. AB - Pancreatic cancer arising in the neck or body of the gland frequently presents at a more advanced stage compared with tumors arising in the pancreatic head. Vascular invasion into the celiac axis is a common contraindication to surgical resection. We report an en bloc pancreatectomy, splenectomy, and total gastrectomy with resection of the celiac axis for a pancreatic body adenocarcinoma invading the celiac axis and posterior stomach. Collateral blood flow to the liver via the gastroduodenal artery was initially identified on preoperative computed tomography (CT) imaging and confirmed intraoperatively with vascular testing before resection. One of 14 regional lymph nodes was positive for malignancy, and surgical margins were free of tumor. The patient's postoperative course was unremarkable. En bloc resection of the celiac axis and hepatic artery may be safely performed in select patients with localized pancreatic cancer invading these vessels. PMID- 15797233 TI - Spontaneous small bowel perforations due to intestinal tuberculosis should not be repaired by simple closure. AB - Intestinal tuberculosis is a major problem in many regions of the world. The incidence of it is rising in Western countries due to immigration from Third World countries and human immunodeficiency virus infection. The difference between the simple closure and resection and anastomosis was evaluated in this study. Retrospectively, 12 patients with intestinal tuberculosis diagnosed histopathologically among 50 patients with free intestinal perforations operated on between 1995 and 2003 at Turgut Ozal Medical Center were evaluated. Each patient underwent routine laboratory tests and radiologic studies. The most common symptoms of patients were abdominal pain, night sweats, and weight loss. Sites of perforation were ileum in 10 patients (multiple perforation in 4) and jejunum in 2 patients (both had multiple perforations). The perforation was closed by primary closure in 7 patients. Resection-anastomosis was performed in 5 patients. Leaks occurred in overall 3 of 7 patients with primary closure. Three of the 7 patients with leaks due to septicemia died. The mortality rate among all patients was 25%. Intestinal tuberculosis should be kept in mind as a cause in free intestinal perforations. Because of high mortality rate, the resection of the affected area and anastomosis may be the treatment of choice rather than primary closure. PMID- 15797232 TI - Effect of "white diet" during bile monitoring with Bilitec 2000 on esophageal pH metry in patients with gastroesophageal reflux disease. AB - With introduction of the Bilitec 2000 device, intraluminal bile monitoring has become a standard technique for evaluation of patients with gastroesophageal reflux disease and symptoms of bile reflux. A specific "white diet" excluding colored food is necessary to provide correct measurements. The influence of this specific diet on simultaneous esophageal pH monitoring is unknown. Forty patients with reflux symptoms were studied prospectively. Meal times and supine and erect phases of measurement were recorded in a standardized fashion using a patient protocol. Esophageal 24-hour pH monitoring with administration of a "colorless" diet (water, milk, potatoes, fish, chicken) was started on day 1, followed by esophageal 24-hour pH-metry with intake of a normal diet on day 2. Data from the two successive pH measurements were compared. The prevalence of a pathologic esophageal pH-metry was significantly higher during intake of a normal diet compared with a colorless diet (P=0.025). During total and upright phases, administration of a white diet led to significant reduction in the percentage of time with a pH less than 4 (P or = 40 microm): the cuticulum/subcuticulum of the inside left lower arm and an area 5 mm behind the left earlobe. After 12 weeks of treatment, patients receiving the homeopathic preparation exhibited an increased number of nodal points, increased flow rates of erythrocytes in both arterioles and venules, increased vasomotion, and a slight reduction in hematocrit vs. baseline. None of these changes were observed in the control group and the differences between treatment groups were statistically significant. Partial oxygen pressure increased significantly in the Vertigoheel group compared with the control group. In addition, in Vertigoheel patients, significantly increased numbers of cell-wall adhering leucocytes were observed, accompanied by increased local concentrations of the adhesion molecules ICAM-1. The microcirculatory changes were associated with a reduction in the severity of vertigo in the actively treated patients, both as assessed by the treating physician and by the patients themselves. The data support a pharmacological effect on microcirculation from the treatment. PMID- 15797257 TI - Effects of a static magnetic field of either polarity on skin microcirculation. AB - Our specific aim was to investigate whether a local static magnetic field of a permanent magnet, of either pole, affects resting skin blood perfusion. This was done by measuring skin blood perfusion (SBF) by laser-Doppler in dorsum skin of 2nd and 4th fingers of the nondominant hands of 12 volunteers. Both fingers were first exposed to sham magnets, and then the 2nd finger was exposed alternately to north and south poles of a neodymium magnet that produced a field of 4024 G at the palmar part of the finger and a field of 879 +/- 52 G at the site of finger dorsum SBF measurement. Each of the three exposure intervals was 15 min. SBF values were analyzed by first computing the average SBF during the last 5 min of each of the three 15-min exposure intervals. These SBF averages were initially tested for magnet or magnet-pole effects by analysis of variance for repeated measures with finger as a factor, using SBF values for each finger as the test variable. Results of this analysis revealed a large variability in finger SBF among subjects and no significant difference in SBF between exposure conditions (P = 0.705) or any significant interaction between SBF and finger (P = 0.396). However, when intersubject variability was reduced by using the flow difference between treated and nontreated fingers in each exposure interval as the test variable, a statistically significant effect (P = 0.016) attributable to magnet exposure was uncovered. This effect was a reduction in resting SBF in the magnet exposed fingers that was similar for north and south pole magnet exposure. The present findings are the first to demonstrate a direct effect of locally applied magnets on human skin blood perfusion. PMID- 15797258 TI - Induction of cyclooxygenase-2 by anandamide in cerebral microvascular endothelium. AB - Anandamide (AEA), an endogenous cannabinoid receptor agonist, is a potent vasodilator in the cerebral microcirculation. AEA is converted to arachidonic acid (AA) by fatty acid amidohydrolase (FAAH), and the conversion of AA to prostaglandins has been proposed as a potential mechanism for the vasodilation. Although AEA stimulated prostaglandin production by mouse cerebral microvascular endothelial cells, no [(3)H]prostaglandins were produced when these cells were incubated with [3H]AEA. Incubation with R(+)-methanandamide (MAEA), a stable analogue of AEA that is not a substrate for FAAH, produced a similar increase in PGE2 production as AEA. The PGE2 production induced by either AEA or MAEA was completely inhibited by NS-398, a selective cyclooxygenase (COX)-2 inhibitor, suggesting that COX-2 was induced. AEA and MAEA increased the expression of COX-2 protein in a time-dependent manner. This increase occurred as early as 1 h and reached maximum at 2 h. Induction of COX-2 protein by AEA was partially inhibited by AM-251, a selective cannabinoid receptor-1 antagonist. Furthermore, AEA increased COX-2 promoter activity approximately twofold above baseline in a fragment ranging from -1432 to +59, the full-length of the COX-2 promoter, and the increase in COX-2 promoter activity produced by AEA was partially inhibited by AM-251. These results indicate that AEA increased COX-2 expression at the transcriptional level through, at least in part, a cannabinoid receptor-1 mediated mechanism in cerebral microvascular endothelium. PMID- 15797259 TI - Comparison of the signaling mechanisms by which VEGF, H2O2, and phosphatase inhibitors activate endothelial cell ERK1/2 MAP-kinase. AB - VEGF-induced ERK1/2 activation is mediated by a signaling mechanism involving the sequential activation of PLCgamma-PKC-Raf1-MEK-ERK1/2. This signaling pathway is necessary, but not sufficient for ERK1/2 activation, as VEGF-induced generation of reactive oxygen species (ROS) is also required. The molecular interaction by which VEGF-induced ROS generation is coordinated with the PLCgamma plus PKC dependent pathway is not certain, and the goal of this study was to clarify this issue. Prior investigations examining ROS-induced signaling have focused on the cellular protein tyrosine phosphatases (PTPs), and we asked whether a PTP participates in ERK1/2 activation in endothelial cells. We show that both the general PTP inhibitor vanadate, and a dominant negative inhibitor of SHP-1, mimics the effects of VEGF in activating ERK1/2. The phosphatase inhibitors induce ERK1/2 activation in endothelial cells lacking VEGF receptors, indicating that the inhibitors target a downstream effector. As is the case after VEGF treatment, the phosphatase inhibitors do lead to the activation of PLCgamma, and a pharmacological inhibitor of the Src kinases blocks this. These results lead to the conclusion that inhibition of a protein tyrosine phosphatase activates endothelial cell ERK1/2 by a signaling mechanism involving the sequential activation of Src-PLCgamma-PKC-Raf1-MEK-ERK1/2. VEGF treatment most likely activates this pathway by inhibiting SHP-1 through a ROS-dependent mechanism. PMID- 15797260 TI - Pigment epithelium-derived factor inhibits oxidative stress-induced apoptosis and dysfunction of cultured retinal pericytes. AB - Pigment epithelium-derived factor (PEDF) is a potent inhibitor of angiogenesis in the mammalian eye, suggesting that loss of PEDF is implicated in the pathogenesis of proliferative diabetic retinopathy. However, a role for PEDF in early diabetic retinopathy remains to be elucidated. Since oxidative stress is thought to be involved in pericyte loss and dysfunction, one of the changes characteristic of early diabetic retinopathy, we investigated whether and how PEDF could protect cultured retinal pericyte against oxidative stress injury. High glucose (30 mM) increased intracellular reactive oxygen species (ROS) generation in pericytes, which was completely blocked by PEDF. High glucose or H2O2 was found to induce growth retardation and apoptotic cell death of pericytes. PEDF completely restored these cytopathic effects on pericytes. An increased ratio of bax to bcl 2 mRNA level with subsequent activation of caspase-3 was observed in high-glucose or H2O2-exposed pericytes, which was also completely prevented by PEDF. PEDF significantly increased glutathione peroxidase (GPx) mRNA levels and activity in pericytes. Further, PEDF was found to completely inhibit high-glucose- or H2O2 induced increase in a mRNA ratio of angiopoietin-2 to angiopoietin-1 and up regulation of VEGF mRNA levels in pericytes. PEDF mRNA levels themselves were down-regulated in high-glucose- or H2O2-exposed pericytes. These results demonstrate that PEDF protects against high-glucose- or H2O2-induced pericyte apoptosis and dysfunction through its anti-oxidative properties via GPx induction. Our present study suggests that substitution of PEDF proteins might be a promising therapeutic strategy for treatment of patients with early diabetic retinopathy. PMID- 15797261 TI - Orally administered lenalidomide (CC-5013) is anti-angiogenic in vivo and inhibits endothelial cell migration and Akt phosphorylation in vitro. AB - The thalidomide analogue and immunomodulatory drug (IMiD) lenalidomide (CC-5013, REVLIMID) is emerging as a useful treatment for a number of cancers and has recently entered phase III trials for multiple myeloma. It has been suggested that the anti-tumor effect of lenalidomide is related to its anti-angiogenic potency. In this regard, we have previously shown that lenalidomide inhibits angiogenesis in both rat and human in vitro models but does not affect endothelial cell proliferation. We now show that oral administration of lenalidomide attenuates growth factor-induced angiogenesis in vivo; the rat mesenteric window assay was utilized to show that lenalidomide significantly inhibits vascularization in a dose-dependent manner. We also found that lenalidomide significantly inhibits growth factor-induced endothelial cell migration. This correlates with the inhibitory effect of lenalidomide on growth factor-induced Akt phosphorylation, thereby providing a potential mechanism for its anti-migratory and subsequent anti-angiogenic effects. These data further support the use of lenalidomide as an orally administered drug for the effective treatment of angiogenesis-dependent conditions, including cancer, and suggest a potential mechanism of action. PMID- 15797262 TI - Increased heme oxygenase-1 expression in infarcted rat hearts following human bone marrow mesenchymal cell transplantation. AB - Heme oxygenase-1 (HO-1) plays an important role in oxidative stress and recent studies indicate that it is a graft survival protein in cardiac and liver transplant models. In this study, we investigated the relation between the expressions of HO-1 and the effects of human bone marrow mesenchymal cells (MSCs) transplantation to xenogenic rat hearts with experimental myocardial infarction (MI). A total of 5 x 10(6) cells in 100 microl PBS or equal volume PBS alone were injected into the ischemic zones immediately post-MI. At 1, 3, and 7 days post MI, cardiac function was evaluated by echocardiography, the expression of HO-1 was assessed by real-time PCR and Western blot, the localization of HO-1 protein was determined under immunofluorescence microscopy. The infarct size was examined by histology. The numbers of Hoechst-33342 positive MSCs were evaluated under immunofluorescence microscopy and also by flow cytometry after isolation from the host hearts. The results indicated that the HO-1 expressions were markedly increased at both mRNA and protein levels in comparison with injection of PBS at each time point post MSCs transplantation (P < 0.01). HO-1 was revealed both in transplanted MSCs and recipient cardiomyocytes by immunofluorescence. Up regulated HO-1 expression was accompanied by increase of the numbers of Hoechst 33342 positive MSCs, the reduction of infarct size, and the improvement of cardiac function. Transplantation of human MSCs could up-regulate HO-1 expression in infarct rat hearts, which might play an important role in protecting transplanted MSCs, cardiomyocytes survival, and cardiac function improvement during the early stage after MI. PMID- 15797263 TI - Intravenous glycine after cecal ligation and puncture has no effect on impaired hepatic microperfusion, leukocyte adhesion, and mortality in septic rats. AB - Recent studies indicated that prefeeding of a glycine supplemented diet reduces the hepatic inflammatory response and liver damage in sepsis. We investigated the effect of a glycine-enriched infusion on hepatic microcirculatory disturbances and mortality in a rat model of sepsis after the onset of the disease. Male Wistar rats (240 +/- 13 g) underwent cecal ligation and puncture (CLP) or laparotomy (LAP). A glycine (CLP + Gly, n = 24), valine (CLP + Val, n = 24), or sodium chlorid (CLP + Sc, n = 24) infusion was started 2 h after CLP. The LAP group received sodium chloride intravenously (LAP + Sc, n = 18 ). Five hours, 10 h, and 20 h after CLP or LAP intravital microscopy (IVM) was performed to investigate leukocyte-endothelial interaction (LEI) and mean erythrocyte velocity in liver sinusoids (sMEV) and postsinosoidal venules (vMEV). The portal blood flow (PBF), hepatic enzyme liberation, and glycine values in blood were measured. Immunohistochemical staining for ICAM-1 in liver tissue was performed and survival was observed. Glycine values were significantly elevated in the CLP + Gly vs. the CLP + Val and the CLP + Sc group at every timepoint of investigation. Glycine infusion had no beneficial effects on sMEV, vMEV, LEI, hepatic enzyme liberation, and survival. Heart rate and mean arterial pressure remained stable but PBF decreased significantly in all groups 20 h after CLP. Although glycine reduces the hepatic inflammatory response and liver damage in pretreatment of septic rats, there was no effect of intravenous glycine after the onset of sepsis in our experiments. Our animal model does not support the use of glycine in patients. PMID- 15797264 TI - The CNS pericyte response to low oxygen: early synthesis of cyclopentenone prostaglandins of the J-series. AB - Exposure to low oxygen induces adaptive changes at the microvascular level that are beneficial to cell survival. These adaptive changes involve complex signaling mechanisms between the vascular endothelial cell and the pericyte, and are important to the maintenance of vascular homeostasis and hemostasis. We have investigated the early response of the central nervous system (CNS) microvascular pericyte to low oxygen. In vitro exposure of primary rat CNS pericytes to low oxygen induced the rapid synthesis and release of the cyclopentenone prostaglandin (PG) PGD2 and PGJ2 within 15-30 min following hypoxic stress signal. Hypoxia-induced release of PGD2/PGJ2 was COX-1 dependent and did not involve COX-2. The exogenous addition of 15-deoxyDelta(12,14) PGJ2 to pericytes under normoxic conditions increased glut-1 protein in the absence of hypoxia. PGD2 and PGJ2 may be early signaling molecules in the pericyte stress response. PMID- 15797265 TI - Possible therapeutic use of vasodilator iontophoresis. AB - BACKGROUND: Investigation into the effects of a novel vasodilator delivery method (for the eventual treatment of scleroderma related digital ulceration) on healthy controls is reported. When Raynaud's phenomenon (episodic cold-induced colour changes of the fingers) occurs in the context of scleroderma, it can be extremely severe, leading to ulceration and sometimes gangrene. The current treatment of choice for scleroderma-related critical digital ischaemia and/or ulceration is intravenous prostanoid therapy, necessitating hospitalisation. However, iloprost is often poorly tolerated and may be ineffective. METHODS: This study utilises a newly designed iontophoresis chamber which has the potential to allow a therapeutic, rather than diagnostic application for vasodilatory iontophoresis. Ten healthy controls underwent whole finger iontophoresis with 1% acetylcholine chloride for 2 min at 100 microA. Iontophoresis with varying treatment times and currents was carried out on a subset of subjects to determine the effect on perfusion increase. RESULTS: A significant increase in perfusion following iontophoresis was found, compared to the adjacent, untreated finger (P < 0.001). Maximum increase as a percentage from baseline, mean [SD] = 100 [66]%. Both treatment time and current have an approximately linear relationship with perfusion increase. CONCLUSIONS: Iontophoresis of the whole finger administers drugs locally with no systemic effects and warrants further investigation as a therapy. PMID- 15797266 TI - Relationship of inspiratory flow rate and volume on digit tip skin and ulnar artery vasoconstrictor responses in healthy adults. AB - Reflex vasoconstrictive responses are commonly used to assess sympathetic nerve function of the extremities. A regularly used reflex trigger is deep and rapid inspiration. However, little is known about the relationship between respiratory effort and the magnitude of the resultant vasoconstrictor response. This study investigates the association of inspiratory flow rate and volume to the level of vasoconstrictor reflexes in the 4th digit tip skin and the ulnar artery in 12 healthy volunteers (age range 21-65, mean 38.1 years). The effects of Peak Inspiratory Flow (PIF) and Forced Inspiratory Vital Capacity (FIVC) on blood flow were measured at three levels of respiratory effort: low (PIF < 1.0 l/min; FIVC < 1.0 l), medium (PIF = 1.0-1.5 l/min; FIVC = 1.0-1.5 l), and high (PIF > 1.5 l/min; FIVC > 1.5 l) at two separate occasions. Ulnar vasoconstrictor responses showed good correlation with all levels of respiratory effort. Skin digit tip responses showed good correlation at medium and high levels but not at low respiratory effort. Repeatability of both tests was good. Correlation of inspiratory flow rate to vasoconstriction was consistently better than with inspiratory volume. Both digit tip and ulnar vasoconstrictor responses were maximal with greatest respiratory effort. PMID- 15797269 TI - Images in resuscitation: Frank Pantridge and the world's first miniature portable defibrillator. PMID- 15797267 TI - Reflex vasoconstrictor responses of the healthy human fingertip skin. Normal range, repeatability, and influencing factors. AB - Skin vasoconstrictor reflexes (SVCRs) are mediated by sympathetic vasoconstrictor nerve fibers and used to test sympathetic function of the extremities. Since there is a dearth of published systematic data on inspiratory gasp-induced SVCR, we studied its range, repeatability, and influencing factors in 64 healthy subjects (mean age 41.9 years, range 20-78 years, 32 males). Mean vasoconstriction for the second digit was 54% (+/-19)/54% (+/-19) (right/left) and 69% (+/-16)/68% (+/-15) for the fourth digit. Forth digit vasoconstriction was significantly larger than the second digit (P < 0.01), but second and fourth digit correlation was good (Pearson: 0.521-0.741). Repeatability using single measure intraclass correlation was good with 0.62/0.69 for the second digit and 0.64/0.80 for the fourth digit (right/left). A trend for decreasing SVCR was seen with increasing age and weight, no effect was seen for gender, height, or blood pressure. The SVCR repeatability and range of variation seen in this study compare favorably with tests of heart rate variability, thermal threshold detection, and motor nerve conduction. PMID- 15797270 TI - The resuscitation greats. Janos Balassa and resuscitation by chest compression. PMID- 15797271 TI - Emergency medical services in Zimbabwe. AB - Emergency medical services in Zimbabwe are of a very variable standard, and exist in many forms: *Reasonably well-developed urban emergency medical services systems mixed with very poorly resourced and under-developed rural services. *Very high patient workloads, with severely ill medical patients and a large proportion of major trauma and multiple-casualty situations (public safety is given a low priority, and public transport is poorly regulated). *Long emergency response times and patient transport distances. *Somewhat under resourced and under developed emergency departments, with large numbers of critically ill acute patients, as well as many non-emergency/chronic patients who have no other access to appropriate health care. This paper provides a description of the development of ambulance services and acute health care in Zimbabwe, and outline the current demands on the system. Particular reference is made to the City of Harare Ambulance Service, which is considered to be the most developed of the local authority services. PMID- 15797273 TI - Long-term effect of introducing an early warning score on respiratory rate charting on general wards. AB - The respiratory rate is an early indicator of disease, yet many clinicians underestimate its importance and hospitals report a poor level of respiratory rate recording. We studied the short- and long-term effects of introducing a new patient vital signs chart and the modified early warning score (MEWS), which incorporates respiratory rate on the prevalence of respiratory rate recording in six general wards of our hospital. Prior to the commencement of the study, the average percentage of occupied beds where at least one respiratory rate recording had been made in a single 24-h period was 29.5+/-13.5%. After the introduction of the new vital signs chart to all six wards, and the introduction of MEWS to three wards, this rose to 68.9+/-20.9%. When all six wards had been using both the new chart and the MEWS system for almost 1 year, the figure had reached 91.2+/-5.6%. During the pre-introduction period, there was no difference in the prevalence of respiratory rate recording between the specialties (orthopaedic, 26.9%; surgery, 32.9%; medicine, 29.8%; p=0.118). During the second two audit periods, the prevalence of respiratory rate monitoring was consistently higher on medical wards than on surgical and orthopaedic wards (p<0.001). The study confirms the long-term beneficial effect of introducing the MEWS system on respiratory rate recording into the general wards of our hospital. As respiratory rate abnormalities are early markers of disease, it is hoped that improved monitoring will have an impact on the nature and timeliness of the response to critical illness. This may have an impact on the future incidence of potentially avoidable cardiac arrest, deaths and unanticipated intensive care unit admission. PMID- 15797274 TI - Differences in time to defibrillation and intubation between two different ventilation/compression ratios in simulated cardiac arrest. AB - OBJECTIVE: During basic life support (BLS) by a two-rescuer-team early defibrillation and ALS procedures should be performed without interruptions of the BLS-ventilation/compression sequence. The objective of this study was to determine the impact of a ventilation/compression ratio of 5:50 versus 2:15 on the time intervals "Start BLS to first shock" and "Start BLS to intubation". METHODS: Using a random cross over design 40 experienced paramedics performed a standard BLS/ALS-algorithm according to ILCOR guidelines in a manikin model with ventricular fibrillation (resusci skillreporter anne, Laerdal, Norway) performing both the 2:15 and the 5:50 ventilation/compression ratio. BLS was started with bag/valve/mask ventilation, a semi-automatic defibrillator (corpuls 08/16S) was connected with the manikin, ECG-analysis and three shocks were performed and the tracheal intubation was prepared. Ventilation/compression sequence was only interrupted during ECG-analysis and defibrillation. Expiratory volumes and number of compressions were measured. Variables were compared using paired Students t test. In addition paramedics were interviewed about work-flow and emotional stress during the tests. RESULTS: The time interval "Start BLS to first shock" was 78 s (2:15-group) versus 63 s (5:50-group), p<0.0001, the time interval "Start BLS to intubation" was 183 s (2:15-group) versus 150 s (5:50-group), p<0.0001, mean ventilation volumes per minute were 4490 ml (2:15-group) versus 4370 ml (5:50-group), p>0.1, mean number of compressions were 65 min-1. (2:15 group) versus 68 min-1 (5:50-group), p>0.1. The work-flow and emotional stress was appraised by the paramedics to be significantly superior in the 5:50 ratio (p<0.0001). CONCLUSIONS: The ventilation/compression ratio of 5:50 compared with 2:15 during BLS with an unsecured airway reduces the time until the first defibrillation and tracheal intubation was performed without changes in ventilation volume and compressions per minute. The Paramedics stated that the 5:50 ratio improved the work-flow and reduced the emotional stress. PMID- 15797272 TI - A systematic review of currently available pharmacological neuroprotective agents as a sole intervention before anticipated or induced cardiac arrest. AB - We conducted a Medline search for controlled studies evaluating currently available drugs for pharmacological neuroprotection. They had to be administered prior to transient global cerebral ischaemia without further non-pharmacological measures. We deliberately excluded focal ischaemia since its pathophysiology is substantially different from global ischaemia. A total of 45 articles conducted exclusively in laboratory animals met these criteria. The following classes of agents were evaluated: anaesthetics, GABAergic drugs, calcium-antagonists, anticonvulsives, sodium-channel blockers, potassium-channel activators, NMDA receptor antagonists, hormones, vasodilators, dopamine- and alpha2-agonists, magnesium, xanthine oxidase- and cyclooxygenase inhibitors, a nootropic, a protease inhibitor, and immunosuppressants. Some of them were applied chronically and others administered via clinically impracticable routes. The available literature favours isoflurane, phenytoin, lamotrigine, magnesium, and potentially, nimodipine, and flunarizine. If factors like costs, toxicity, side effects, route and mode of application are considered, isoflurane and MgSO4 that have also been safely applied to patients with compromised left ventricular pump function are advantageous but their true role in human neuroprotection remains unclear. PMID- 15797275 TI - Outcome after cardiac arrest: predictive values and limitations of the neuroproteins neuron-specific enolase and protein S-100 and the Glasgow Coma Scale. AB - BACKGROUND AND PURPOSE: Patients resuscitated from cardiac arrest are at risk of subsequent death or poor neurological outcome up to a persistent vegetative state. We investigated the prognostic value of several epidemiological and clinical markers and two neuroproteins, neuron-specific enolase (NSE) and S-100 protein (S-100), in 97 patients undergoing cardiopulmonary resuscitation (CPR) after non-traumatic cardiac arrest between 1998 and 2002. RESULTS: 52.6% of the patients died, 28.8% survived with severe, moderate or without neurological disorders, and 18.6% remained in a persistent vegetative state. Unconsciousness>48 h after CPR predicted a 60.6-fold (95% CI 14.3287-257.205, p=0.001) and a Glasgow Coma Scale (GCS)<6 points after 72 h a 11.2-fold (CI 95%, 3.55-36.44, p<0.001) risk of poor neurological outcome. Serum levels>or=65 ng/ml for NSE and >or=1.5 microg/l for S-100 increased the risk of death and persistent vegetative state 16.8 (95% CI 2.146-131.520)- and 12.6 (95% CI 1.1093-99.210) fold, respectively. By combination of the GCS with elevated serum concentrations of both neuroproteins above the cut off levels on third day after CPR a poor neurological outcome was predicted with a specificity of 100%. CONCLUSION: The combination of GCS with the serum levels of both neuroproteins at 72 h after CPR permit a more reliable prediction of outcome in post arrest coma than the single markers alone, independent of the application of anaesthetic agents. PMID- 15797277 TI - Pre-randomization and de-randomization in emergency medical research: new names and rigorous criteria for old methods. AB - Clinical trials are performed to determine if a therapy is effective in the treatment of a disease. The methods of randomization and blinding are used to assure that the only planned difference between the two groups is the therapy itself, and differences in outcome cannot be attributed to bias. Emergency medical conditions, and in particular therapies that must be administered in an emergency, present challenges to inclusion, exclusion, randomization, and blinding that are at times insurmountable in the context of available resources. Pre-randomization (that is, assigning the therapy to be used before the event occurs) and de-randomization (that is, removing randomized cases that do not meet established inclusion criteria) may address some of the challenges resulting from emergency enrollment but have the potential to create bias. We describe these techniques, and provide criteria that should be employed if pre-randomization and/or de-randomization are being considered. It is possible to use these techniques to successfully complete clinical trials that would not have been possible using only standard methodology and still ensure that results are without bias. PMID- 15797276 TI - Prospective use of a clinical decision rule to identify pulmonary embolism as likely cause of outpatient cardiac arrest. AB - A clinical decision rule (CDR) derived retrospectively found that 57% of outpatients aged 65 years or less, with witnessed arrest+PEA had pulmonary embolism (PE) as cause of cardiac arrest. These retrospectively studied patients also had significant frequency of pre-arrest respiratory distress, altered mental status, and shock. OBJECTIVES: (1) To test prospectively the feasibility and diagnostic accuracy of this CDR. (2) To test if the pre-arrest clinical triad of respiratory distress, altered mental status and shock predicts the presence of PE. All EMS personnel (N=204) in an urban EMS system and Emergency Department physicians (N=143) at 7 hospitals were included in the CDR and data collection. INCLUSION CRITERIA: age 18-70, non-trauma, witnessed arrest, PEA as the first and primary rhythm. Exclusion: defibrillation before or more often than once after PEA. Criterion standards: autopsy or predefined cardiopulmonary imaging for PE. Over 21 months, 44 subjects were enrolled. Thirty-three subjects had a criterion standard (N=20 autopsy, 13-other criteria). 18/33 (54%; 95% CI 36-72%) had PE. Of the PE arrests, 88% were witnessed by EMS (N=8) or ED physicians (N=8), compared with 47% in the non-PE group (N=3 EMS and N=4 ED). Of the PE arrests, 83% had at least two of the three components of the triad versus 33% of the non-PE group (95% CI for difference 20-79). Mortality was 100% in the PE group. Analysis of the EMS cardiac arrest registry indicated that 65% of all patients served by the EMS system, ageor=70 mmHg). Liver dysoxia was monitored as increase in liver surface pCO2 during phases I and II. Observation phase III was to 72 h. RESULTS: During HS, preventing a decrease in MAP below 40 mmHg required HHS 4.9+/-0.6 ml/kg (all data mean+/-S.E.M.), compared to LR 62.2+/-16.6 ml/kg (P<0.001), with no group difference in MAP. Uncontrolled blood loss during HS from the tail stump was 13.3+/-1.9 ml/kg with HHS infusion, versus 12.6+/-2.5 ml/kg with LR infusion (P=0.73). Serum sodium concentrations were moderately elevated at the end of HS in the HHS group (149+/-3 mmol/l) versus the LR group (139+/-1 mmol/l) (P=0.001), and remained elevated throughout. Liver pCO2 increased during HS in both groups equally (P<0.001 versus baseline), and tended to return to baseline levels at the end of HS. Blood gas and lactate values throughout did not differ between groups. During HS, 2 of 10 rats in the HHS group versus 0 of 10 in the LR group died (P=0.47). There was no difference between HHS and LR groups in survival rates to 72 h (3 of 10 in the HHS group versus 2 of 10 in the LR group) (P=1.0). Survival times, by life table analysis, were not different (P=0.75). CONCLUSION: In prolonged uncontrolled HS, a titrated i.v. infusion of HHS can maintain controlled hypotension with only one-tenth of the volume of LR required, without increasing blood loss. This titrated HHS strategy may not increase the chance of long-term survival. PMID- 15797282 TI - Bilateral tension pneumothorax. AB - The report discusses three patients who presented with pulseless electrical activity (PEA), caused by chronic respiratory disease, with bilateral tension pneumothorax. In each case needle decompression failed to relieve the tension and cardiac output was restored only after the insertion of a chest tube. PMID- 15797283 TI - Diagnostic pitfalls in patients with hypoxic brain damage: three case reports. AB - OBJECTIVE: Our aim was to assess the possible diagnostic pitfalls in three patients with hypoxic brain damage who had partly conflicting clinical, biochemical, and electrophysiological data and were in a persistent vegetative state (PVS) following cardiac arrest (CA). METHODS: Serum concentrations of the destruction proteins, neuron-specific enolase (NSE) and protein S-100B (S-100B), were measured on days 1-3, and 7; somatosensory evoked potentials (SEPs) were recorded within 48 h and on day 7 after CA. RESULTS: Two patients had significantly increased concentrations of NSE and S-100B during the first 3 days after CA, a finding that indicates ongoing neuronal destruction. In contrast, the SEPs of these patients were normal or showed only a diminished amplitude configuration. In the third patient the SEPs demonstrated a bilateral loss of cortical responses repeatedly, but both destruction proteins were only slightly above the upper normal values on all study days. CONCLUSION: Our findings demonstrate that a poor prognosis can only be established if either SEPs, NSE, or S-100B are very abnormal. The conflicting results in our patients indicate that variable values may reflect different patterns of neuropathological damage caused by diffuse hypoxia. We, therefore, favour a multi-modal approach with a combination of clinical, biochemical, and electrophysiological investigations in order to predict neurological outcome after CA reliably. PMID- 15797284 TI - Successful resuscitation with mechanical CPR, therapeutic hypothermia and coronary intervention during manual CPR after out-of-hospital cardiac arrest. AB - A 62-year-old man suffered out-of-hospital cardiac arrest and was treated with mechanical compression-decompression during transport to the hospital. In the emergency department, 28 min after cardiac arrest, spontaneous circulation returned briefly but the patient rapidly became asystolic and mechanical compression-decompression was again applied. After further resuscitation a spontaneous circulation returned and the patient was transferred, deeply comatose, to the coronary intervention laboratory while therapeutic hypothermia was induced. In the laboratory the heart arrested again and coronary angiography was performed during manual CPR revealing a left main stem occlusion. After successful reperfusion of the heart the patient was transferred to the intensive care unit with an intra-aortic balloon pump. The patient was treated with hypothermia for 24 h and awoke without neurological sequelae after a sustained intensive care period of 13 days. The present case is an example of how modern resuscitation principles implementing new clinical and experimental findings may strengthen the chain of survival during resuscitation. PMID- 15797286 TI - Two pairs is better than one -- double gloves at cardiac arrests. PMID- 15797287 TI - Radio-histological correlation: a key to a clear understanding of images in breast pathology. PMID- 15797285 TI - Elevated serum beta-D-glucan level and depressed neutrophil phagocytosis in a heatstroke patient. AB - Endotoxemia has been reported as a mechanism for the fatal sequela after heatstroke. Subsequent disseminated fungal infection in a heatstroke patient has been also described. Beta-D-glucan, a constituent of the fungal cell wall, is an early diagnostic measure for fungal infection. In a heatstroke case, we examined for the first time levels of serum beta-d-glucan and endotoxin. A 34-year-old man with a body temperature of 43.5 degrees C was admitted in a state of shock. Prior to the development of disseminated intravascular coagulopathy (DIC), a remarkable elevation of serum beta-D-glucan level to 116 pg/mL (normal level<6.0 pg/mL) was revealed on the first day of admission. However, serum endotoxin was not detected when using a method that excluded beta-D-glucan contamination from endotoxin measurement (normal level<1.0 pg/mL). This change of beta-D-glucan level was accompanied by a depressed neutrophil function, especially in phagocytosis of 34% (normal range 70-90%) but not in bacterocidal function (81% versus a normal range of 70-100%). After intensive care including continuous hemodiafiltration, the patient regained consciousness but remained ataxic due to cerebellar infarction, which might have resulted from DIC, and subsequent bilateral fungal oculitis were revealed 45 days after admission. This case report demonstrates the elevation of serum beta-D-glucan but normal endotoxin levels after heatstroke, which may prompt further study to re-examine the serum levels of endotoxin in such catastrophic insults. PMID- 15797288 TI - Quiz case. Breast sarcoidosis presenting as a metastatic breast cancer. AB - A case of sarcoidosis presenting initially as a breast mass with subclinical pulmonary and medullary involvement highly mimicking metastatic breast carcinoma is reported. The mammographic, ultrasound and CT scan findings are described with a review of the literature. PMID- 15797290 TI - Use of radiology for the pathologist in the management of breast lesions. AB - Today radiology is an essential step in the pathological analysis of breast biopsies. It is determinant at each stage of the management of non palpable lesions, clusters of microcalcifications and opacities, whether this concerns the needle biopsy or the surgical excision. Firstly, an X-ray is necessary to ensure that the core needle biopsy specimen has been adequately sampled and when samples with microcalcifications are selected by the radiologist, management can be more specific and accurate. In the case of surgical specimens, the X-ray confirms the presence of the radiographic abnormality or the clip indicating the site of the surgical excision which guides sampling. Some radiographic features also provide information on underlying pathologies allowing management to be adapted accordingly. Radiographs are also important to ensure that microscopically detected microcalcifications or lesions exactly correspond to the radiographic abnormality in size and location. The paraffin block can also be X-rayed to select those containing microcalcifications for additional slicing. It is also important to identify the presence of modifications caused by the core needle biopsy (fibrosis, haemorrhage and inflammation) and to carefully recognize displacement of epithelial cells and pseudo-emboli resulting from the needle procedure. Such correlation between radiology and pathology is essential so that appropriate management of the specimen can be adapted and to avoid pitfalls arising from pre-operative procedures. PMID- 15797289 TI - Normal and pathological breast, the histological basis. AB - Breast tissue is heterogeneous, associating connective and glandular structures, which grow and change cyclically under hormonal regulation. Hormones are also thought to be the main determinant of the major benign and malignant pathologies encountered in the breast. Benign lesions are more frequent and fibrocystic changes are by far the most common among them. They usually associate different entities, (adenosis, fibrosis, cysts and hyperplasia) but vary in intensity and extension. Thus, their clinical and radiographic presentation is extremely different from one patient to another. Adenofibroma is the most frequent tumour. It also undergoes modifications according to hormonal conditions. About 90% of malignant tumours are primary carcinoma. The incidence of intra-ductal carcinoma has risen dramatically since the development of screening because of its ability to induce calcification. Two mechanisms could be involved in the formation of calcification: one active (tumour cell secretion of vesicles), the other passive (necrotic cell fragments are released). Invasive carcinoma comprises numerous histological types. Stromal reactions essentially determines their shape: a fibrous reaction commonly found in ductal carcinoma creates a stellate lesion while other stroma, inflammatory (medullary carcinoma), vascular (papillary carcinoma) or mucinous determine nodular lesions whose borders push the surrounding tissue. The histological features which give rise to the radiographic pattern will be emphasised. PMID- 15797291 TI - The normal breast and its variations in mammography. AB - The mammographic appearance of the breast varies along the lifetime due to physiological modifications or use of hormonal therapies. Density of the glandular tissue is due to amount of cellular elements of the gland and to hydratation of the tissues. Normal variations are encountered as for example breast asymmetry. The currently breast composition should be described with the BI-RADS lexicon classification. Mammary asymmetry is frequent and has to be differentiated from pathologic changes. A good mammographic technique is mandatory for an adequate visualisation of the breast tissues. PMID- 15797292 TI - Stellate images: anatomic and radiologic correlations. AB - The stellate images are the most well-known and most typical finding in the diagnosis of breast cancer. Its positive predictive value is very high. However, in some cases, the stellate images can correspond to benign lesions. Stellate images require high quality mammograms, precise analysis, and as always in senology, confirmation by clinical examination, ultrasound if necessary, and in the case of palpable lesions, cytology. In this work, we will study malignant and benign stellate images mammographic-pathologic correlation as well as the importance of stellate images in the detection of non-palpable lesions. The typical stellate finding is correlated with the phenomena of fibrosis and elastosis and it is impossible to distinguish benign spicules from malignant spicules on mammography. We will present guidelines in the face of stellate image. Stellate images are for the most part suggestive of malignant lesions, and their discovery should lead to suspicion of cancer until the contrary is proven. PMID- 15797293 TI - Mammography of ductal carcinoma in situ of the breast: review of 909 cases with radiographic-pathologic correlations. AB - We retrospectively analysed mammographies of 909 ductal carcinoma in situ (DCIS) (1980-1999) and compared our results to those of literature. Microcalcifications were present in 75% of the cases, and soft-tissue abnormalities in 27% cases with association with calcifications in 14% of cases. Palpable masses were found in 12% of the cases and nipple discharge was present in 12% of the cases. The radiographic-pathologic correlation allowed to suspect the DCIS "aggressiveness" on radiologic signs. Granular, linear, branching and/or galactophoric topography of the microcalcifications were correlated with necrosis, grade 3, comedocarcinoma type. A number of microcalcifications higher than 20 was correlated with necrosis and grade 3. Mammographic size was correlated to histologic size. Masses were correlated with grade 1. A diagnosis strategy can be proposed with a multidisciplinar approach. PMID- 15797294 TI - Precursor lesions of invasive breast cancer. AB - The increasing application of mammography, mainly in screening programs for the early detection of breast cancer, and the high technical standard of imaging has resulted in the detection of clinically occult breast tumors. Considering that only diagnosis at an early stage will be able to change the prognosis of breast cancer, this diagnostic challenge appears to be the most exciting field in both breast imaging and breast pathology. Especially the precursor lesions need to be diagnosed and defined precisely to understand their prognostic significance. In imaging, the morphologic appearance of precursor lesions is usually neither typical nor pathognomonic. They have to be assessed histologically using percutaneous interventions. Recent molecular studies have demonstrated various genetic alterations in the ductal epithelium, with the earliest onset in atypical ductal hyperplasia. The recent WHO classification, which is based on molecular data and histopathological features, attempts to define in particular the precursor lesions and low grade intraductal carcinomas. The clinical importance of the various grades has to be assessed. Intimate cooperation between diagnostic radiologist and pathologist is essential. PMID- 15797295 TI - Dynamic optical breast imaging: a new technique to visualise breast vessels: comparison with breast MRI and preliminary results. AB - The purpose of our study was to explore and evaluate an innovative imaging approach, which consists on imaging the breast parenchyma by means of photoluminescence detectors (LED) and analysis of dynamic data. Breast magnetic resonance imaging (MRI) was chosen as the reference imaging method, as this is considered to be nowadays the gold standard for breast vascularisation evaluation. Preliminary results reveal a good correlation between breast MRI findings and light images. PMID- 15797296 TI - Significance analysis of qualitative mammographic features, using linear classifiers, neural networks and support vector machines. AB - Advances in modern technologies and computers have enabled digital image processing to become a vital tool in conventional clinical practice, including mammography. However, the core problem of the clinical evaluation of mammographic tumors remains a highly demanding cognitive task. In order for these automated diagnostic systems to perform in levels of sensitivity and specificity similar to that of human experts, it is essential that a robust framework on problem specific design parameters is formulated. This study is focused on identifying a robust set of clinical features that can be used as the base for designing the input of any computer-aided diagnosis system for automatic mammographic tumor evaluation. A thorough list of clinical features was constructed and the diagnostic value of each feature was verified against current clinical practices by an expert physician. These features were directly or indirectly related to the overall morphological properties of the mammographic tumor or the texture of the fine-scale tissue structures as they appear in the digitized image, while others contained external clinical data of outmost importance, like the patient's age. The entire feature set was used as an annotation list for describing the clinical properties of mammographic tumor cases in a quantitative way, such that subsequent objective analyses were possible. For the purposes of this study, a mammographic image database was created, with complete clinical evaluation descriptions and positive histological verification for each case. All tumors contained in the database were characterized according to the identified clinical features' set and the resulting dataset was used as input for discrimination and diagnostic value analysis for each one of these features. Specifically, several standard methodologies of statistical significance analysis were employed to create feature rankings according to their discriminating power. Moreover, three different classification models, namely linear classifiers, neural networks and support vector machines, were employed to investigate the true efficiency of each one of them, as well as the overall complexity of the diagnostic task of mammographic tumor characterization. Both the statistical and the classification results have proven the explicit correlation of all the selected features with the final diagnosis, qualifying them as an adequate input base for any type of similar automated diagnosis system. The underlying complexity of the diagnostic task has justified the high value of sophisticated pattern recognition architectures. PMID- 15797297 TI - Computer-aided detection (CAD) in mammography: does it help the junior or the senior radiologist? AB - OBJECTIVES: To evaluate the impact of a computer-aided detection (CAD) system on the ability of a junior and senior radiologist to detect breast cancers on mammograms, and to determine the potential of CAD as a teaching tool in mammography. METHODS: Hundred biopsy-proven cancers and 100 normal mammograms were randomly analyzed by a CAD system. The sensitivity (Se) and specificity (Sp) of the CAD system were calculated. In the second phase, to simulate daily practice, 110 mammograms (97 normal or with benign lesions, and 13 cancers) were examined independently by a junior and a senior radiologist, with and without CAD. Interpretations were standardized according to BI-RADS classification. Sensitivity, Specificity, positive and negative predictive values (PPV, NPV) were calculated for each session. RESULTS: For the senior radiologist, Se slightly improved from 76.9 to 84.6% after CAD analysis (NS) (one case of clustered microcalcifications case overlooked by the senior radiologist was detected by CAD). Sp, PPV and PNV did not change significantly. For the junior radiologist, Se improved from 61.9 to 84.6% (significant change). Three cancers overlooked by the junior radiologist were detected by CAD. Sp was unchanged. CONCLUSION: CAD mammography proved more useful for the junior than for the senior radiologist, improving sensitivity. The CAD system may represent a useful educational tool for mammography. PMID- 15797298 TI - Breast radiological cases: training with BIRADS classification. AB - The American College of Radiology has established guidelines for outcomes monitoring known as the Breast Imaging Reporting and Data System. The last edition of the BIRADS classification includes mammography, but also ultrasonography and MRI. Radiologists must be used to the BIRADS lexicon and the BIRADS assessment, in order to clarify mammography reports and to facilitate communication with the other physicians. This work is a training on 20 mammography cases to be familiar with the BIRADS classification. PMID- 15797299 TI - Visualization of intracranial vessel anatomy using high resolution MRI and a simple image fusion technique. AB - A new technique for fusion and 3D viewing of high resolution magnetic resonance (MR) angiography and morphological MR sequences is reported. Scanning and image fusion was possible within 20 min on a standard 1.5 T MR-scanner. The procedure was successfully performed in 10 consecutive cases with excellent visualization of wall and luminal aspects of the intracranial segments of the internal carotid artery, the vertebrobasilar system and the anterior, middle and posterior cerebral artery. PMID- 15797300 TI - In vivo 1H MR spectroscopy of thyroid carcinoma. AB - To determine if proton magnetic resonance spectroscopy (1H MRS) of thyroid carcinoma is feasible and to determine if 1H MRS spectra of malignant tumors differ from that of normal thyroid tissue. We performed 1H MRS at 1.5 T at echo times (TE) 136 and 272 ms to examine eight patients with thyroid cancer (primary tumour or nodal metastasis) larger than 1 cm3 in size and five volunteers with normal thyroids. Spectra acquired from six primary tumors (three anaplastic carcinomas, two papillary carcinomas and one follicular carcinoma) and two nodes (two papillary carcinoma metastases) were analyzed in the time-domain using a non linear least squares fitting algorithm with incorporation of prior knowledge. Choline (3.2 ppm) was identified in all solid carcinomas with a mean choline/creatine of 4.3 at TE 136 ms and 5.4 at TE 272 ms. Ratios for malignant tumors at TE 136 ms ranged from 1.6 in well differentiated follicular carcinoma to 9.4 in anaplastic carcinoma. No choline was detected in normal thyroid tissues. Our results showed that 1H MRS is a feasible technique for the evaluation of malignant thyroid tumors larger than 1 cm3 and that proton spectra of malignant tumors differ from that of normal thyroid tissue. PMID- 15797301 TI - Role of computed tomography in the follow-up of hepatic and peritoneal metastases of GIST under imatinib mesylate treatment: a prospective study of 54 patients. AB - PURPOSE: To prospectively analyze the evolution of hepatic and peritoneal unresectable metastases from gastrointestinal stromal tumors (GIST) under imatinib mesylate, a new targeted treatment, which induces changes in lesion structure. MATERIALS AND METHODS: 54 patients with metastases from GIST underwent an abdominal and pelvic computed tomography examination without and with contrast enhancement, before and during treatment with imatinib mesylate. The number and size of lesions and contrast enhancement were noted before treatment and every 2 weeks for the first 2 months, then every 2 months for the first year of treatment and every 3 months thereafter. RESULTS: 27 patients presented with both hepatic and peritoneal metastases, 14 had only peritoneal and 13, only hepatic disease. On baseline imaging, all metastases were hypodense heterogeneous lesions with progressive, concentric enhancement. After treatment (mean duration of follow-up: 23 months) metastases decreased in size number and enhancement in 35/54 patients, remained stable in 2 patients and increased in 14 patients. In 13/39 patients with hepatic metastases a cyst-like appearance was noted. Reactivation after a partial response appeared first as a focal, peripheral, solid nodule in the wall of a cystic lesion, or an increase in lesion density, before size regrew. CONCLUSION: Besides the classic size criterion, a decrease in density and in contrast enhancement with stable "near cystic" lesions signifies a good response. A more aggressive approach (surgery or radiofrequency ablation) may be indicated for initially focal recurrences with a stable size. PMID- 15797302 TI - Parametric and quantitative analysis of MR renographic curves for assessing the functional behaviour of the kidney. AB - The aim of this study was to refine the description of the renal function based on MR images and through transit-time curve analysis on a normal population and on a population with renal failure, using the quantitative model of the up-slope. Thirty patients referred for a kidney MR exam were divided in a first population with well-functioning kidneys and in a second population with renal failure from ischaemic kidney disease. The perfusion sequence consisted of an intravenous injection of Gd-DTPA and of a fast GRE sequence T1-TFE with 90 degrees magnetisation preparation (Intera 1.5 T MR System, Philips Medical System). To convert the signal intensity into 1/T1, which is proportional to the contrast media concentration, a flow-corrected calibration procedure was used. Following segmentation of regions of interest in the cortex and medulla of the kidney and in the abdominal aorta, outflow curves were obtained and filtered to remove the high frequency fluctuations. The model of the up-slope method was then applied. Significant reduction of the cortical perfusion (Qc = 0.057+/-0.030 ml/(s 100 g) to Qc = 0.030 +/- 0.017 ml/(s 100 g), P < 0.013) of the medullary perfusion (Qm = 0.023 +/- 0.018 ml/(s 100 g) to Qm = 0.011 +/- 0.006 ml/(s 100 g), P < 0.046) and of the accumulation of contrast media in the medulla (Qa = 0.005 +/- 0.003 ml/(s 100 g) to Qa = 0.0009 +/- 0.0008 ml/(s 100 g), P < 0.001) were found in presence of renal failure. High correlations were found between the creatinine level and the accumulation Qa in the medulla (r2 = 0.72, P < 0.05), and between the perfusion ratio Qc/Qm and the accumulation Qa in the medulla (r2 = 0.81, P < 0.05). No significant difference was found in times to peak between both populations despite a trend showing Ta the time to the end of the increasing contrast accumulation period in the medulla, arriving later for renal failure. Advances in MR signal calibration with the building of quantitative model such as the up-slope allow to assess kinetic and haemodynamic and functional parameters of the diseased kidney. PMID- 15797303 TI - Percutaneous uterine artery embolization for the treatment of symptomatic fibroids: current status. AB - Uterine artery embolization (UAE) is increasingly being used as an alternative treatment to hysterectomy for symptomatic fibroids. Symptoms of pelvic pressure, urinary frequency and menorrhagia are controlled in 73-98% of patients who undergo UAE. At the 1-year follow-up, the uterus may shrink by up to 55% but re growth of fibroid may however occur. The rate of major complications and amenorrhoea following this procedure is low, ranging in most series from 1 to 3.5% and 1 to 7%, respectively. Nevertheless, the rate of amenorrhoea in women over 45 seems to be higher. In order to completely block the arterial supply to the fibroid, UAE is typically performed in both uterine arteries. Different embolic agents are used such as polyvinyl alcohol, gelfoam and more recently gelatine tris-acryl microspheres. After UAE, perfusion of the uterus is maintained. Uterine function is therefore conserved and although women who become pregnant after UAE seem to be at risk for malpresentation, pre-term birth, cesarean delivery and postpartum hemorrhage, successful pregnancies after UAE have been reported in some series. A major technical problem with UAE remains the possible presence of fibroid blood supply from other sources, such as the ovarian arteries or other pelvic branches, which can lead to failure of the procedure. In conclusion, although randomized trials are still underway, UAE appears a good option for those patients who whish to conserve their fertility or when surgery is contra-indicated. However, to evaluate the long-term effects of UAE longer follow up is required. PMID- 15797304 TI - The value of contrast enhanced power Doppler ultrasonography in differentiating hypoehoic lesions in the peripheral zone of prostate. AB - OBJECTIVE: To characterize the hypoechoic prostate nodules in the peripheral zone by means of power Doppler ultrasonography (PDUS) and contrast enhanced PDUS (CE PDUS). MATERIALS AND METHODS: Thirty-two patients with dijital rectal examination findings suspicious for malignancy and/or a serum PSA level higher than 4 ng/dl, and who had hypoechoic nodules on transrectal ultrasonography were enrolled in this prospective study. Power Doppler ultrasonography (PDUS) images before and after sonocontrast (Levovist) injection were photographed for further evaluation. All patients were also evaluated by a systematic and nodule targeted biopsy approach. The photographs were re-evaluated by two radiologists for the presence and type of vascularity. The type of vascularity was categorized as peripheral, central, mixed and penetrating. The latter two were accepted as representatives of malignancy. The results of PDUS and CE-PDUS were compared to each other and to the pathological results. RESULTS: There was a high level of interobserver agreement (Kappa: 0.80-0.95). Nineteen patients, but only 14 nodules were malignant on pathological examination. Sensitivity, specificity positive and negative predictive values for PDUS were 57, 50, 47, and 60%, respectively. On CE PDUS, the sensitivity increased (93%) in expense of specifity (17%). The positive and negative predictive values were 46 and 75%, respectively. There was no significant difference between PDUS and CE-PDUS. CONCLUSION: This study revealed that both benign and malignant nodules might be hypervascular and show malignant type of vascularity on PDUS. Contrary to some recent reports, we can conclude that the CE-PDUS does not provide a considerable aid to the diagnosis of prostate carcinoma on morphological basis due to its very low specificity despite its relatively higher sensitivity. PMID- 15797305 TI - Distress and expression of natural killer receptors on lymphocytes. AB - Chronic distress has been associated with alterations in natural killer (NK) cell and T cell percentages and function. NK cells express inhibitory and stimulatory receptors that regulate cytotoxicity and cytokine secretion. T cells and T cells expressing CD56 (NKT cells) also express these NK-associated receptors, which are thought to serve the same function. The objective of this study was to examine the relationship between distress and expression of NK-associated receptors on NK cells, T cells, and NKT cells. Using multicolor flow cytometry and validated questionnaires, we studied twenty-nine healthy adults with a bimodal age spread. Whereas distress was related to significantly lower percentages of CD3(+) T cells, it was related to significantly higher percentages of NKT cells. Distress was associated with significantly higher percentages of T cells expressing NK associated receptors including CD94 and KIR. In contrast, distress was associated with significantly lower percentages of NK cells bearing KIR (GL183) receptors. Our findings suggest a possible role for NK-associated receptors in distress related alterations in lymphocyte maturation, trafficking, or activity. PMID- 15797307 TI - Stress relief by oxygen? PMID- 15797306 TI - Optimism and immunity: do positive thoughts always lead to positive effects? AB - The effects of dispositional optimism, as defined by generalized positive expectations for the future, on physical health are mixed, especially in diseases that can be immunologically mediated such as HIV and cancer. Both experimental and naturalistic studies show that optimism is negatively related to measures of cellular immunity when stressors are difficult (e.g., complex, persistent, and uncontrollable) but positively related when stressors are easy (e.g., straightforward, brief, and controllable). Although the negative relationship between optimism and immunity has been attributed to the violation of optimists' positive expectancies and subsequent disappointment, empirical evidence suggests that it is more likely to be a consequence of optimists' greater engagement during difficult stressors. For example, negative mood does not account for the effect, but conscientiousness, a personality facet related to engagement, does. The mixed immunological correlates of optimism may explain why it does not consistently predict better disease outcomes. PMID- 15797308 TI - Stress is not associated with thyroid peroxidase autoantibodies in euthyroid women. AB - OBJECTIVE: Multiple genes and environmental factors play a role in the etiology of autoimmune thyroid disease (AITD). In Graves' hyperthyroidism, stress is such an environmental factor, but whether it plays a role in Hashimoto's hypothyroidism is unknown. We used validated questionnaires to evaluate an association between TPO antibodies, an early marker for AITD, and self-reported stress. SUBJECTS AND METHODS: Recently Experienced Stressful Life Events, Daily Hassles, and mood (tendency to report positive and negative affects) were assessed in 759 euthyroid subjects. RESULTS: TPO antibodies were found in 183/759 (24%) of subjects. The TPO-Ab positive subjects were older (39.7+/-12 vs. 34.2+/ 12 years; p<.001) than the TPO-Ab negative subjects, but the number of daily hassles (24+/-14 vs. 25+/-14; p=.24), the number of stressful life events (10+/-6 vs. 11+/-6; p=.09), and the scores on the affect scales (22.1+/-7.4 vs. 22.2+/ 7.3; p=.89 for negative affect and 38.2+/-5.1 vs. 38.3+/-5.3; p=.91 for positive affect) were similar in TPO-Ab positive and TPO- Ab negative subjects. CONCLUSIONS: We found no association between recently experienced stressful life events, daily hassles or mood and the presence of TPO antibodies in these euthyroid women. PMID- 15797309 TI - Impaired wound contraction and delayed myofibroblast differentiation in restraint stressed mice. AB - Previous research has shown that psychological stress delays wound closure by >25%. Gene expression of pro-inflammatory cytokines and the maturation of the epithelium were also impaired by stress (Mercado et al.). Wound contraction contributes to the speed of wound closure (Hunt and Hopf). In the current study, wound contraction was decreased by >45% (p<.01) in restraint stressed mice. Fibroblast migration and differentiation into smooth muscle alpha-actin (SmalphaA) -expressing myofibroblasts were delayed in RST mice through day 7 post wounding. In addition, there was a 25 (p<.01), 48 (p<.01), and 38% (p<.05) decrease in SmalphaA mRNA levels at days 1, 3, and 5 post-wounding in RST mice, respectively. Cytokines that regulate fibroblast migration and differentiation include transforming growth factors-beta1, -beta2, and -beta3 (TGF-betas). Although expression of TGF-beta1 mRNA was downregulated by >25% (p<.01) in RST mice on day 3 post-wounding, no significant differences were detected in active or total TGF-beta1 protein levels. Stress did not alter the expression of TGF beta2 or -beta3 through day 5 post-wounding. Thus, these data indicate that stress delays wound contraction and myofibroblast differentiation, which are likely independent of expression of TGF-beta1, -beta2, and -beta3. PMID- 15797310 TI - Hyperbaric oxygen therapy ameliorates stress-impaired dermal wound healing. AB - Psychological stress has been shown to dysregulate healing in both humans and animals. Studies indicate the possibility for decreased oxygen supply, and increased oxygen demand, in the wounds of the stressed animals. Oxygen is an important mediator of wound healing, and its availability can limit healing rate. Hence, in a mouse model of stress-impaired healing, the hypothesis that hyperbaric oxygen therapy would ameliorate the effect of stress on dermal wound healing was tested. Hyperbaric oxygen therapy (HBO) twice a day during early wound healing significantly ameliorated the effects of stress, bringing healing to near-control levels. There was no significant effect of HBO on the wounds of control animals. Wound inducible nitric oxide synthase (iNOS), modulated by psychological stress and oxygen balance, was studied for gene expression by real time PCR. Expression of iNOS increased in stressed mice on days 1 (205%; p<.0001), 3 (96%; p<.03), and 5 (249%; p<.03), post-wounding. HBO treatment of the stressed animals decreased iNOS expression by 62.6% (p< .02) day 1 post wounding. There was no significant effect of HBO on wound healing and iNOS expression in the control animals. Methods aimed at increasing tissue oxygenation, like HBO, have a high therapeutic potential. Their molecular mechanisms, implicated in wound healing, elude clarification due to the lack of appropriate animal models. Our current findings represent the first experimental evidence, demonstrating that HBO corrects stress-impaired dermal wound healing. PMID- 15797311 TI - Transgenic overexpression of interleukin-1 receptor antagonist in the CNS influences behaviour, serum corticosterone and brain monoamines. AB - The effects of brain-directed overexpression of human soluble interleukin-1 receptor antagonist (hsIL-1ra) on behaviour, serum corticosterone (CST) levels and concentrations of dopamine (DA), 5-hydroxytryptamine (5-HT) and their metabolites in different brain regions, were investigated in six months old homozygotic transgenic male mice (Tg hsIL-1ra(+/+)). The transgenic and age matched wild type (WT) mice were subjected to a battery of behavioural tests for analysis of open field (OF) behaviours, anxiety in elevated plus maze (EPM), and motor performance in rotarod. One week after the last behavioural test, half of the mice from each genotype were subjected to a mild stress, while the remaining mice served as controls for the determination of serum CST levels and monoamine concentrations in different brain regions. Tg hsIL-1ra(+/+) mice had higher locomotor scores and showed less habituation in the OF test, spent more time in the open arms of the EPM and had similar motor performance as compared to WT mice. The serum CST levels were comparable, both in basal conditions and upon stress, in the two genotypes. Tg hsIL-1ra(+/+) mice had lower concentrations of DA, 5-HT and their metabolites in several brain regions, with different effects on monoamine turnover upon stress. In conclusion, brain-directed overexpression of hsIL-1ra resulted in increased locomotion and decreased habituation, an anxiolytic effect, but did not influence motor performance. Finally, the activation of hypothalamo-pituitary-adrenal (HPA)-axis was comparable in the two genotypes, however Tg hsIL-1ra(+/+) mice had a modified metabolism of brain monoamines as compared to WT mice. PMID- 15797312 TI - Epstein-Barr virus shedding by astronauts during space flight. AB - Patterns of Epstein-Barr virus (EBV) reactivation in 32 astronauts and 18 healthy age-matched control subjects were characterized by quantifying EBV shedding. Saliva samples were collected from astronauts before, during, and after 10 space shuttle missions of 5-14 days duration. At one time point or another, EBV was detected in saliva from each of the astronauts. Of 1398 saliva specimens from 32 astronauts, polymerase chain reaction analysis showed that 314 (23%) were positive for EBV DNA. Examination by flight phase showed that 29% of the saliva specimens collected from 28 astronauts before flight were positive for EBV DNA, as were 16% of those collected from 25 astronauts during flight and 16% of those collected after flight from 23 astronauts. The mean number of EBV copies from samples taken during the flights was 417 per mL, significantly greater (p<.05) than the number of viral copies from the preflight (40) and postflight (44) phases. In contrast, the control subjects shed EBV DNA with a frequency of 3.7% and mean number of EBV copies of 40 per mL of saliva. Ten days before flight and on landing day, titers of antibody to EBV viral capsid antigen were significantly (p<.05) greater than baseline levels. On landing day, urinary levels of cortisol and catecholamines were greater than their preflight values. In a limited study (n=5), plasma levels of substance P and other neuropeptides were also greater on landing day. Increases in the number of viral copies and in the amount of EBV specific antibody were consistent with EBV reactivation before, during, and after space flight. PMID- 15797314 TI - Stress exposure modulates peptidergic innervation and degranulates mast cells in murine skin. AB - Stress is said to induce itchiness of the skin, exacerbate inflammatory skin diseases, and inhibit wound healing. Neuropeptides such as substance P (SP) may play a role in these processes. Recently, we were able to show that both stress or SP are associated with neurogenic inflammation and increased apoptosis in the murine hair follicle. Moreover, peptidergic cutaneous innervation is subject to lifelong plasticity due to its association with the cyclic growth of hair follicles. However, peripheral neuronal plasticity has never been reported in altered interactions between the nervous and immune systems under perceived stress. Here, we show for the first time plasticity of the cutaneous peptidergic innervation in response to stress. After exposure to sonic stress, the number of SP+ nerve fibers in the back skin of C57BL/6 mice with their hair follicles in the resting phase of the hair cycle (telogen-low numbers of nerve fibers) increased significantly. Such nerve fibers contacted mast cells more frequently. At the same time, the percentage of degranulated mast cells increased significantly associated with a rise in apoptotic cells in the skin. Increased numbers of peptidergic nerve fibers correlated with increased numbers of growth associated protein 43 (Gap-43)+ nerve fibers, which is a marker for growing nerves. Thus, neuronal plasticity and increased neuro-immune interaction occur under stress and may alter inflammatory skin diseases and trophic functions in the skin where neurogenic inflammation plays a part. PMID- 15797313 TI - Preliminary evidence for lymphocyte distribution differences at rest and after acute psychological stress in PTSD-symptomatic women. AB - This study investigated circulating natural killer (NK), CD4+ and CD8+ cells in response to acute psychological challenge among mothers of child cancer survivors with and without posttraumatic stress symptoms (PTSS). Control mothers of healthy children (n=9) were compared to 17 cancer mothers with (PTSS: n=9) and without PTSS (No PTSS: n=7) under conditions of rest, after a generic stressor (MAT: mental arithmetic task) and a personalized stressor (script-driven trauma imagery), and after recovery from each stressor. Results indicate the PTSS group had higher percentage CD4+ and lower CD8+ levels than non-symptomatic women and blunted NK reactivity to generic challenge. Multiple regression analyses indicated PTSS effects were independent of self-reported distress. Contrary to expectations, cancer mothers without PTSS were not significantly different from controls on tonic or phasic immune outcomes. Also unlike predictions, reactivity to challenge was greatest to the non-social MAT stressor compared to the personalized challenge for all groups. Conclusions are constrained by study limitations (e.g., small sample size and potential phase order effects). Nonetheless, results are consistent with an emerging literature on PTSS associated immune differences and further suggest these effects may be distinct from that associated with subjective distress more generally. PMID- 15797315 TI - Sensitivity of ex situ and in situ spectral surface plasmon resonance sensors in the analysis of protein arrays. AB - We have investigated the sensitivity of ex situ (analysis under air condition) and in situ (analysis under liquid condition) spectral SPR sensors, which were self-constructed with fiber optic spectrometers. The sensitivity of SPR sensors was analyzed in the wavelength range of 550-780 nm by the interactions of streptavidin and biotinylated IgG, and the sensitivity was dependent on the wavelength of measurements. The sensitivity of an ex situ SPR sensor operated at the long wavelength range from 712 nm was approximately 2.6 times higher than that at the short wavelength range from 571 nm. In addition, the sensitivity of an ex situ spectral SPR sensor was about twice as high as that of an in situ spectral SPR sensor for the same resonance wavelength range. This was interpreted in that the difference in sensitivity between two SPR sensors was significantly caused by the evanescent field intensity at the metal/dielectric interface. Thus, it was suggested that ex situ spectral SPR sensors operated at the long wavelength range are sensitive biosensors for the high-throughput analysis of protein interactions on protein arrays. PMID- 15797316 TI - Molecularly imprinted ligand-exchange recognition assay of glucose by quartz crystal microbalance. AB - Molecular imprinted polymers (MIP) as a recognition element for sensors are increasingly of interest and MIP-quartz crystal microbalance (QCM) have started to appear in the literature. In this study, we have combined quartz crystal microbalance with MIP to prepare a sensor using the ability of glucose to chelate of copper (II) ion of methacrylamidohistidine (MAH) monomer to create ligand exchange (LE) assembled monolayer which is suitable for glucose determination. The study includes the measurement of binding interaction of molecularly imprinted QCM sensor via ligand interaction, investigation of the pH effect on frequency shift and recognition selectivity studies of glucose-imprinted polymer with respect to methyl-alpha-d-glucopyranoside and sucrose. Bmax (number of binding sites) and K(D) (dissociation constant of the metal-chelate copolymer) were also calculated using Scathard plot and the detection limit was found as 0.07 mM. MIP showed higher glucose-binding affinity than a well-known glucose binding protein, conconavalin A. PMID- 15797317 TI - A compact CMOS biochip immunosensor towards the detection of a single bacteria. AB - Recent use of biological warfare (BW) agents has led to a growing interest in the rapid and sensitive detection of pathogens. Therefore, the development of field usable detection devices for sensitive and selective detection of BW agents is an important issue. In this work, we report a portable biochip system based on complementary metal oxide semiconductor (CMOS) technology that has great potential as a device for single-bacteria detection. The possibility of single bacteria detection is reported using an immunoassay coupled to laser-induced fluorescence (LIF) detection. Bacillus globigii spores, which are a surrogate species for B. anthracis spores, were used as the test sample. Enzymatic amplification following immunocomplex formation allowed remarkably sensitive detection of B. globigii spores, and could preclude a complicated optical and instrumental system usually required for high-sensitive detection. Atomic force microscopy (AFM) was employed to investigate whether B. globigii spores detected in the portable biochip system exist in single-cell or multicellular form. It was found that B. globigii spores mostly exist in multicellular form with a small minority of single-cell form. The results showed that the portable biochip system has great potential as a device for single-particle or possibly even single organism detection. PMID- 15797318 TI - Nanogold particle-enhanced oriented adsorption of antibody fragments for immunosensing platforms. AB - A general design strategy for immunosensing platforms has been proposed on the basis of Nanogold particle-enhanced oriented adsorption of antibody fragments. Quartz crystal microbalance (QCM) as a model transducer was fabricated with plasma-polymerized film (PPF) of n-butyl amine and then with nanogold particles resulting in a PPF-nanogold adsorption procedure for half-IgG fragments obtained by reduction of intact immunoglobulin (IgG). Thermodynamic studies reveal that the proposed procedure is superior to the traditional oriented ones in that it created immunosurface of increased antibody surface density (amount) and antigen binding constants. Sensors produced according to the new immobilization procedure exhibit better immunosensing performances including high sensitivity, fast response rate, and favorable operational stability etc. This Nanogold particle enhanced immobilization technique may be tailored as a promising alternative for various immunosensing platforms in solid-phase immunoassay and affinity chromatography. PMID- 15797319 TI - A low-cost, portable generic biotoxicity assay for environmental monitoring applications. AB - Fish chromatophores have been shown to be promising biosensors for the detection of hostile agents in the environment. However, state-of-art methods for such applications are still based on extensive use of data/signal processing, in conjunction with need for a skilled human observer to carry out the detection. As a result, conventional methods are complex, costly and cumbersome rendering them useless for field applications requiring low-cost portable solutions capable of fast detection. A new technique is proposed based on the popular scheme of observing the aggregation response in chromatophores for detection of toxicity, and a solution using optical detection and electronic processing is outlined. This scheme has the advantage of being low in cost while providing simple, fast and reliable detection. PMID- 15797320 TI - A novel electrochemical synthesis of poly-3-methylthiophene-gamma-cyclodextrin film Application for the analysis of chlorpromazine and some neurotransmitters. AB - Gamma-cyclodextrin is an eight glucose unit conical structure, which possesses a hydrophobic internal cavity exhibiting supramolecular complexing properties and a hydrophilic exterior due to the presence of hydroxyl groups. In this work, we have synthesized for the first time a functionalized stable film of poly-3 methylthiophene combined with gamma-cyclodextrin (P3MT/gamma-CD) in tetrabutylammonium hexafluorophosphate/acetonitrile solution. A potentiostatic mode was employed for the film growth. Cyclic voltammetry and electrochemical impedance spectroscopy were used for the characterization of the new film. The resulting conducting polymer sensor offers interesting analytical performances such as: (a) fast and linear responses towards the neurotransmitters dopamine and l-dopa towards the neuroleptic chlorpromazine with low detection limits of 2 x 10(-7), 10(-6), and 10(-7)M, respectively, and (b) simultaneous detection and well-resolved signals between the compounds of interest and ascorbic acid. To the best of our knowledge, these LOD are among the lowest found in the literature. Optimization of parameters such as interference effect, perm-selectivity, and mechanical stability of the sensor are discussed. PMID- 15797321 TI - Adenoviral p53 effects and cell-specific E7 protein-protein interactions of human cervical cancer cells. AB - We investigated the time-course tumor growth suppression effects of recombinant adenovirus expressing p53 on human cervical cancer cells and cell-specific E7 protein-protein interactions in cell lysates using surface plasmon resonance (SPR) biosensor. Six HPV-infected human cervical cancer cell lines (HPV 16 positive cells, CaSki and SiHa cells; HPV 18-positive cells, HeLa and HeLaS3 cells; and HPV negative C33A and HT3 cells) were used. After infection with AdCMVp53, the cell-specific growth inhibition was studied in vitro and in vivo. Also, we produced the recombinant E7 oncoprotein of HPV 16 type and tested chip based protein-protein interactions with each cell lysate. For each cervical cancer cell, differential cell growth inhibitions were shown via cell count assay and MTT assay. Note that the same trend in suppression levels was shown in CaSki, HeLa and in SiHa, HeLaS3, respectively. In contrast, infection with AdCMVLacZ showed increased cell growth in a manner similar to the negative control group. The levels of p53 protein were notably expressed in CaSki and HeLa more than in SiHa and HeLaS3 for 4 days. In contrast, p53 expression was continually maintained in C33A and HT3 for 6 days. After transfection AdCMVp53 into CaSki- and SiHa-xenografted nude mice, the size of tumor was remarkably decreased in SiHa cells as compared to AdCMVLacZ transfection. The SPR sensor surface was successfully modified with the recombinant E7 oncoprotein and showed cell specific interactions between E7 and its target proteins from cell lysates. The anti-tumor effects were accomplished via differential role of p53-specific apoptotic cell death, which is dependent upon the cervical cancer cell line. Also, a molecular level understanding of cell-dependent protein interaction effects of recombinant E7 was shown. PMID- 15797322 TI - A novel continuous subcutaneous lactate monitoring system. AB - A novel continuous lactate monitoring system has been developed modifying the GlucoDay portable medical device (A. Menarini Diagnostics), already present in the European market, and used to continuously measure glucose levels. Lactate oxidase based biosensors have been developed immobilising the enzyme on nylon net and placing it on a Pt electrode. The biosensor was connected to the portable device provided with a micro-pump and coupled to a microdialysis system. It is capable to record subcutaneous lactate every 3 min. In vitro analytical results confirmed that the sensors respond linearly in the interval of concentration between 0.1 and 10 mmol/L, covering the whole physiological range. During prolonged monitoring periods, the response of the biosensors remained stable, showing a limited drift of 8%, within 60 h. Stability tests are still on route. However, preliminary results have shown a shelf life of about 10 months. In vivo experiments performed on healthy rabbits have demonstrated the good accuracy and reproducibility of the system. A correlation coefficient equal to 0.9547 (N=80) was found, which represents a good correlation between the GlucoDay and the laboratory reference analyser. A 16 h in vivo monitoring on a healthy volunteer has been also performed. PMID- 15797324 TI - A new recombinant cell-based bioluminescent assay for sensitive androgen-like compound detection. AB - A public concern is continuously arising about the presence of natural and anthropogenic compounds which affect human health by modulating normal endocrine functions. These substances, defined as endocrine disrupting compounds (EDC) represent an heterogeneous class of molecules either steroidal or not, sharing the ability of interfering with the endocrine system via nuclear receptor signaling pathways. Therefore there is an urgent need for high throughput screening systems able to detect EDCs and evaluate their biological activity. However, little attention has been dedicated to the development of assays for androgen-like compounds. The present work describes the development and optimization of a new rapid and sensitive bioluminescent yeast-based bioassay for androgen-like compounds in a 96-well microplate format. The bioassay is based on recombinant Saccharomyces cerevisiae cells modified to express human androgen receptor (hAR) and containing the sequence androgen response element (ARE) which drives the expression of Photinus pyralis luciferase, used as reporter gene. A recombinant yeast strain constitutively expressing luciferase was used as external control to correct the light signal accordingly to cell viability and sample matrix aspecific effects. The bioassay responds to testosterone as reference androgen in a concentration-dependent manner from 0.05 to 1000 nM allowing an accurate and precise quantitative evaluation in aqueous environmental samples down to 10(-11)mol/L. Other known androgen-like compounds exhibit similar dose-response behavior, thus permitting the use of the bioassay for an overall detection of androgen-like effect in environmental samples. PMID- 15797323 TI - Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence. AB - A hybridisation-based genosensor was designed on a 100 nm sputtered gold film. This material worked as an immobilisation and transduction surface. A 30-mer sequence that encodes a short lysine-rich region, unique to SARS (severe acute respiratory syndrome) virus, was chosen as target. A complementary strand (probe), labelled with a thiol group at the 3'-end, was immobilised on the film. After blocking the surface, hybridisation with the biotin-conjugated SARS strand (at the 3'-end) took place. Interaction with alkaline phosphatase-labelled streptavidin permits amplified indirect electrochemical detection. The analytical signal is constituted by an electrochemical process of indigo carmine, the soluble product of the enzymatic hydrolysis of 3-indoxyl phosphate. The use of a sensitive electrochemical technique such as square wave voltammetry allowed a detection limit of 6 pM to be obtained for this DNA sequence, lower than any other found in the bibliography. The parameters affecting the methodology were studied, with special attention being placed on selectivity. Specificity was clearly enhanced when interaction time and stringency (in the form of formamide percentage) were increased. With 1h of strand interaction and employing 50% of formamide in the hybridisation buffer, a 3-base mismatch strand was perfectly distinguished from the complementary. PMID- 15797325 TI - Biosensors based on acrylic microgels: a comparative study of immobilized glucose oxidase and tyrosinase. AB - Acrylic microgels are proposed as enzyme immobilizing support in amperometric biosensors. Two enzymes, glucose oxidase and tyrosinase, were entrapped in this matrix and their behaviour is compared. The optimum cross-linking of the polymeric matrix required to retain the enzyme, and to allow the diffusion of the substrate is different for each enzyme, 3.2% for glucose oxidase and 4.5% for tyrosinase. The effect of pH and temperature on the biosensor responses has been studied by experimental design methodology and predictions have been compared with independently performed experimental measurements. A quadratic effect of the variables studied (pH and T) on the biosensor response and the small or null interaction between them was confirmed. The pH results obtained with both methods are coincident revealing an reversible effect on the enzyme. However, the temperature optimum value obtained by experimental design was 10 degrees C lower as a result of an activity decay due to irreversible thermal denaturation of both enzymes. PMID- 15797326 TI - Immuno-capture of Cryptosporidium parvum using micro-well array. AB - A glass slide and micro-well array chip on which anti-Cryptosporidium parvum antibody was immobilized were used for the rapid capture and detection of C. parvum. Biotinylated anti-C. parvum antibodies were spotted onto the streptavidin coated glass slides. C. parvum oocysts were captured specifically on the spot when more than 73 ng of anti-C. parvum antibody was applied onto the glass slide. However, C. parvum oocysts captured on the glass slide were detached by repeating washing steps. To improve the capture efficiency of oocysts, capture was performed in a micro-well format consisting of 1024 wells/2.5 mm2 (32 x 32 wells) fabricated as a chip by photolithography. Instead of a flat surface on a glass slide, each well was 30 microm in diameter and 10 microm in depth. Streptavidin was also immobilized onto the micro-well array. The biotinylated anti-C. parvum antibodies were immobilized efficiently onto the chip using a buffer containing 20% methanol. Using this technique C. parvum oocysts were stably captured onto the chip after repeated washing procedures. These data show that the newly designed micro-well array technique described here is useful for antibody mediated C. parvum capture. PMID- 15797327 TI - Application of a taste evaluation system to the monitoring of Kimchi fermentation. AB - As an objective method, taste evaluation with an instrument is able to supplement the subjective sensory evaluation and to be applied to the optimization of food processing. Kimchi, a Korean traditional pickle fermented with lactic acid bacteria, is expanding its consumption worldwide. The fermentation control of it has been routinely done by measuring titratable acidity and pH. In this study, an eight-channel taste evaluation system was prepared, followed by an application to the monitoring of Kimchi fermentation. Eight polymer membranes which individually responded to cationic or anionic substances were prepared by mixing electroactive materials such as tri-n-octylmethylammonium chloride, bis(2-ethylhexyl)sebacate as the plasticizer and polyvinyl chloride in the ratio of 1:66:33. Each membrane prepared was separately installed onto the sensitive area of an ion-selective electrode to produce the respective taste sensor. The eight-channel sensor array and a double junction reference electrode were connected to a 16-channel high input impedance amplifier. The amplified sensor signals were stored to a personal computer via a multi-channel A/D converter. Two sensor groups composed of the cation-selective and anion-selective polymer membrane electrodes showed characteristic concentration-dependency to various artificial taste substances. As a whole, the response potentials of the sensor array increased during the fermentation period at 4, 10 and 25 degrees C. Even the response potentials of the anion-selective taste sensors slightly increased possibly due to the protonation of anions by liberated H+ ions, thereby leading to a decrease in the anion concentration. When the signal data were interpreted by principal component analysis (PCA), the first PC at 4 degrees C explained most of the total data variance. A close correlation was found between the values of titratable acidity and the first PC, which indicated a possible applicability of the multi-channel taste sensor of this study to the process monitoring of various pickle. PMID- 15797330 TI - High efficiency electrochemical immuno sensors using 3D comb electrodes. AB - To realize highly sensitive electrochemical immunoassays, a micro-fabricated three-dimensional (3D) electrode was fabricated and applied to enzyme immuno assay based on production of a redox species. The dimensions of the electrodes are 10 microm in width and 30 microm in height, with 20 microm spacing in between, and the 30 pairs of anode and cathode electrodes made up a single sensor. This structure lead to enhancement of the electrochemical reaction, nearly 100% of trap ratio of redox species. It can be applied to highly sensitive enzyme immuno sensing based on p-aminophenylphosphate (PAPP). Applicability of this technique to the immuno assay for one of the clinical diagnostic marker proteins (alpha-fetoprotein; AFP) from 6 to 500 ng/mL was demonstrated. PMID- 15797328 TI - Protein array consisting of sol-gel bioactive platform for detection of E. coli O157:H7. AB - Sol-gel-derived bioactive platform was fabricated for detection of pathogenic microbes, E. coli O157:H7. Design flexibility of sol-gel technique and ease of fabrication can fulfill to create the surfaces with structural and chemical features that are compatible with biomaterials such as antibody, enzymes, etc. In this study, the bioactive platform was prepared based on the silica gels, which were produced by hydrolyzing tetraethylorthosilane (TEOS) in ethanol. The mercaptopropyl triethoxysilane (MPTS) was mixed with the TEOS solution for the surface functionalization of bioactive platform. During TEOS hydrolysis, the modified thin film was prepared by sol-gel dip coating. Antibody against E. coli O157:H7 was immobilized with a configuration of protein array using piezo-type dispensing system. Surface morphology of the prepared bioactive platform was analyzed using atomic force microscopy (AFM). The antibody-antigen interaction was investigated with fluorescence microscopy and sandwich type immunoassay using fluorescein isothiocyanate (FITC)-labeled antibody. The results showed that antibody was sequestered within the sol-gel-derived bio-gel due to physical adsorption. The measurement of E. coli O157:H7 was done using the fabricated antibody surface. The fluorescence intensity was proportional to the concentration of E. coli O157:H7, of which the detection limit was 10(2)CFU/ml. PMID- 15797329 TI - Cell immobilization using self-assembled synthetic oligopeptide and its application to biological toxicity detection using surface plasmon resonance. AB - The immobilized cell using self-assembled synthetic oligopeptide was applied to the biological toxicity detection of environmental pollutant. Thin films based on cysteine-terminated synthetic oligopeptides were fabricated for the immobilization of Escherichia coli O157:H7 on gold (Au) substrate. Layer formation and immobilization of E. coli O157:H7 were investigated with surface plasmon resonance (SPR) and atomic force microscopy (AFM). Experimental results showed that the thin film of cysteine-terminated synthetic oligopeptide was successfully fabricated and it could be applied for the immobilization of E. coli O157:H7. The attached living cell was exposed to toxic chemical such as phenol, which induced the change of SPR angle. As the exposed concentration of phenol was increased, the change of plasmon resonance angle was increased, which indicates the decrease of cell viability. The detection limit based on SPR was determined as 5 ppm. The proposed cell immobilization method using self-assembly technique can be applied to construct the cell microarray for the diagnosis, drug detection, and on-site monitoring. PMID- 15797331 TI - A novel optical biosensor format for the detection of clinically relevant TP53 mutations. AB - The TP53 gene has been the subject of intense research since the realisation that inactivation of this gene is common to most cancer types. Numerous publications have linked TP53 mutations in general or at specific locations to patient prognosis and therapy response. The findings of many studies using general approaches such as immunohistochemistry or sequencing are contradictory. However, the detection of specific mutations, especially those occurring in the structurally important L2 and L3 zinc binding domains, which are the most common sites of TP53 mutations, have been linked to patient prognosis and more strongly to radiotherapy and chemotherapy resistance in several major cancers. In this study, the TI-SPR-1 surface plasmon resonance system and Texas Instruments Spreeta chips were used to develop a DNA biosensor based on thiolated probes complementary to these domains. The sensors were able to detect these mutations in both oligonucleotides and PCR products with normal and mutant TP53 DNA, but the difference in hybridisation signal was small. Preliminary experiments to enhance the signal using Escherichia coli mismatch repair proteins, MutS and single strand binding protein were carried out. It was found that MutS was unable to bind to mismatch oligonucleotides, but single strand binding protein was able to bind to single stranded probes, which had not hybridised to the target, resulting in a three-fold increase in the sensitivity of the biosensor. While further work needs to be carried out to optimise the system, these preliminary experiments indicate that the TI-SPR-1 can be used for the detection of clinically relevant mutations in the TP53 gene and that the sensitivity can be increased significantly using single strand binding protein. This system has a number of advantages over current mutation detection technologies, including lower cost, ease of sensor preparation and measurement procedures, technical simplicity and increased speed due to the lack of need for gel electrophoresis. PMID- 15797332 TI - Development of a microfabricated disposable microchip with a capillary electrophoresis and integrated three-electrode electrochemical detection. AB - We have developed microsystems with a capillary electrophoresis and an electrochemical detector. The microfabricated CE-ECD systems are adequate for a disposable type and the characteristics are optimized for application in electrochemical detection. The system was realized by means of a polydimethylsiloxane (PDMS)-glass chip and an indium tin oxide electrode. The injection and separation channels were produced by relatively simple and inexpensive methods. A capillary electrophoresis and a three-electrode electrochemical detector were fabricated on the same substrate with the same fabrication procedure. We measured electropherograms for the testing analytes consisting of catechol and dopamine with different concentrations of 1mM and 0.1mM, respectively. The results showed an efficient and rapid separation and detection of all compounds within a very short time of around 80s using a separate electric field 60 V/cm. We could also successfully achieve an electropherogram of the separation of the 1 kb DNA ladder (8.4 ng/mul) from the 500 bp to 10 kb DNA fragments within just 150 s. PMID- 15797333 TI - Insulator semiconductor structures coated with biodegradable latexes as encapsulation matrix for urease. AB - A new urea biosensor for clinical applications was obtained by immobilization of urease within different latex polymers functionalized by hydroxy, acetate and lactobionate groups. Responses of these biosensors based on pH-ion-selective field effect insulator-semiconductor (IS) systems to urea additions were evaluated by capacitance measurements. UV-visible spectroscopy was used to check the urease activity in various matrixes. A good retention of the catalytic urease activity in the case of the cationic polymers was observed. In addition, rotating disk electrode experiments were carried out to determine the matrix permeability characteristics. Under optimal conditions, i.e. buffer capacity corresponding to 5 mM phosphate buffer, the urea enzyme insulator semiconductor (ENIS) sensors showed a linear response for urea concentrations in the range 10(-1.5) to 10( 4)M. Furthermore, kinetic parameters for the immobilized urease were obtained from Lineweaver-Burk plot. Clearly, a fast response and a good adhesion for the urease-acetate polymer composite films, prepared without using glutaraldehyde as cross-linking agent was observed. PMID- 15797334 TI - Acetylecholinesterase-based biosensor electrodes for organophosphate pesticide detection. II. Immobilization and stabilization of acetylecholinesterase. AB - The dry and wet stability of Drosophila acetylcholinesterase non-covalently immobilized onto polyethyleneimine modified screen-printed carbon electrodes was improved when compared to non-immobilized acetylcholinesterase, and acetylcholinesterase covalently immobilized onto dialdehyde and polyethyleneimine modified electrodes. Stabilizer mixtures were characterized for additional stabilization of acetylcholinesterase during storage in the dry state, with dextran-sulphate/sucrose and polygalacturonic acid/sucrose mixtures proving highly effective for long-term storage of biosensor electrodes. PMID- 15797335 TI - Improved selectivity and stability of glucose biosensor based on in situ electropolymerized polyaniline-polyacrylonitrile composite film. AB - A new type of in situ electropolymerization method was used for electrochemical biosensor design. The biologic film was prepared by in situ electropolymerization of aniline into microporous polyacrylonitrile-coated platinum electrode in the presence of glucose oxidase. The novel glucose biosensor exhibited good selectivity, sensitivity and stability, which showed no apparent loss of activity after 100 consecutive measurements and intermittent usage for 100 days with storage in a phosphate buffer at 4 degrees C. Blood glucose determinations agreed well with standard hospital laboratory analysis. The construction and operational parameters of the biosensor were also optimized. PMID- 15797336 TI - Multiple wavelength fluorescence enhancement on glass substrates for biochip and cell analyses. AB - In microarrays experiments, a serious limitation is the unreliability of low signal intensities data and the lack of reproducibility for the resulting ratios between samples and controls. Most of the light emitted by a fluorophore at the air/glass interface of a glass slide is absorbed by the glass so just a part of the emitted fluorescence is detected. To improve the sensitivity of the fluorescence detection of both common fluorophores Cy3 and Cy5 in DNA microarrays and fluorescent cell analyses, we have designed a multi layer mirror with alternative thin layers of SiO2 and HfO2. This mirror (MOTL) prevents fluorescence absorption, allows the simultaneous enhancement of the fluorescence signals and increases the dynamic range of the slides. Using MOTL slides, Cy3 and Cy5 intensities are enhanced by 5-8-fold, consequently, the fluorescence analysis becomes easier and should allow the detection of low copy number genes or weakly fluorescent cells. With the same approach, other multiple optical thin layer slides could be designed for other series of fluorophores, extending the field of their applications. PMID- 15797337 TI - A glucose biosensor based on electrodeposition of palladium nanoparticles and glucose oxidase onto Nafion-solubilized carbon nanotube electrode. AB - Electrodeposition was used for the co-deposition of glucose oxidase (GOx) enzymes and palladium nanoparticles onto a Nafion-solubilized carbon nanotube (CNT) film. The co-deposited Pd-GOx-Nafion CNT bioelectrode retains its biocatalytic activity and offers an efficient oxidation and reduction of the enzymatically liberated H2O2, allowing for fast and sensitive glucose quantification. The combination of Pd-GOx electrodeposition with Nafion-solubilized CNTs enhances the storage time and performance of the sensor. An extra Nafion coating was used to eliminate common interferents such as uric and ascorbic acids. The fabricated Pd-GOx-Nafion CNT glucose biosensor exhibits a linear response up to 12 mM glucose and a detection limit of 0.15 mM (S/N = 3). PMID- 15797338 TI - Genetically engineered acetylcholinesterase-based biosensor for attomolar detection of dichlorvos. AB - The design of a biosensor for the detection of dichlorvos at attomolar levels is described based on a highly sensitive double mutant (E69Y Y71D) of the Drosophila melanogaster acetylcholinesterase (Dm. AChE). This enzyme has a k(i) for dichlorvos equal to 487 microM(-1)min(-1), which is 300 and 20,000 times higher than that of the wild type Dm. AChE and the Electrophorus electricus AChE (E.el. AChE), respectively. The enzyme is immobilized into microporous-activated conductive carbon, and is used as such for the development of an inhibitor electrochemical biosensor. This E69Y Y71D mutant enables the decrease in the detection limit of the biosensor down to 10(-17) M, which is five orders of magnitude lower compared to the Electropharus electricus-based biosensor and eight orders of magnitude lower than the biosensors described so far. PMID- 15797339 TI - Towards a polymeric binding mimic for cytochrome CYP2D6. AB - A series of fluorescent molecularly imprinted polymers has been prepared with a view to generating material capable of mimicking the binding characteristics of the metabolically important cytochrome isoform CYP2D6. Such polymers would have the possibility to form the sensing element in a high-throughput assay for the prediction of CYP2D6 affinity. The imprinted polymers possessed binding-dependent fluorescence. They re-bound their templates and various cross-reactivities were encountered for test compound/drug recognition. One polymer in particular exhibited a rational discrimination amongst the related synthetic templates and was reasonably successful in recognising CYP2D6 substrates from a drug panel. PMID- 15797340 TI - Optimization of the binding properties of a synthetic anion receptor using rational and combinatorial strategies. AB - The solution conformations of tetrameric and hexameric cyclopeptides containing alternating L-proline and 6-aminopicolinic acid subunits strongly depend on solvent polarity. Whereas in polar solvents, such as d6-DMSO, both peptides prefer on average symmetric conformations with converging NH groups, in less polar chloroform intramolecular hydrogen bonds to the peptide NH groups stabilize other, and in the case of the hexapeptide, non-symmetrical conformations. Independent of the solvent, both peptides interact with anions via their NH groups but whereas anion binding requires a cleavage of the intramolecular hydrogen bonds accompanied by a conformational reorganization in chloroform, in polar solvents the peptides are already well preorganized for anion complexation. Complex formation between anions and the cyclic hexapeptide was even detected in highly competitive D2O/CD3OD or H2O/CH3CN mixtures, which was attributed to the special sandwich-type structure of the complexes formed. Stabilizing these 2:1 aggregates by covalently linking two cyclopeptide rings together affords ditopic receptors with a high anion affinity in protic solvents. Complex stability depends on the structure of the linker with which the two receptor moieties are connected and even more potent anion receptors were obtained by a dynamic combinatorial optimization of this linking unit. PMID- 15797342 TI - Reinvention of the food guide pyramid to promote health. PMID- 15797341 TI - An integrated approach for the design and synthesis of oligonucleotide probes and their interfacing to a QCM-based RNA biosensor. AB - The quantitative determination of specific cellular messenger-RNA is extremely important both in basic and applied research, especially in diagnostic and pharmacological fields. In order to perform a direct and easy quantification of transcripts on cell extracts, the feasibility of an analytical device able to selectively detect a defined target RNA in a complex mixture while avoiding labelling, retrotranscription and amplification steps, has been explored. In particular, several aspects necessary to obtain good selectivity in target recognition, stability, reusability and sensitivity of a gene specific biosensor were considered. For the development of suitable probe-receptors, analysis of the nucleotide sequence of the target mRNA was carried out to localise the preferred binding regions. As criteria for optimisation, we selected accessibility and uniqueness. Oligonucleotide probes, designed to specifically bind these sequences, were synthesised by using particular monomers producing nuclease resistant RNA strands with high affinity towards the target. Quartz crystal microbalance (QCM) technology was selected to realise a microgravimetric sensor able to bind the RNA under investigation through a complementary oligonucleotide probe. Covalent immobilisation of bioreceptor molecules to the transducer sensitive surface ensured a stable integration between the two. The binding ability of immobilised probes was tested evaluating their annealing behaviour with both complementary oligonucleotides and full-length target mRNA. The conditions necessary for the regeneration of biosensor were also assessed. Measurements of shift in QCM resonant frequency, performed by hybridisation experiments in liquido, demonstrate that a label-free RNA-biosensor with high specificity, reusability and the ability to give quantitative information, can be realised. PMID- 15797343 TI - Plant pigments: properties, analysis, degradation. PMID- 15797344 TI - Chitin, chitosan, and co-products: chemistry, production, applications, and health effects. PMID- 15797345 TI - A review of the application of sourdough technology to wheat breads. PMID- 15797346 TI - Detection of insect infestation in stored foods. PMID- 15797347 TI - Compression and compaction characteristics of selected food powders. PMID- 15797348 TI - Safety of newer generation anti-epileptic drugs in non-accidental overdose: an Irish population study. AB - Seven new anti-epileptic drugs (AEDs) have become available in Ireland over the last 10 years; data from animal models and clinical trials suggest that they have a superior safety profile to older AEDs. A specific relationship between epilepsy and psychiatric co-morbidity has long been recognised, including the relationship between epilepsy and suicide. AEDs are common agents taken in intentional drug overdoses. We undertook a study to review the frequency and outcome of non accidental overdose with seven new AEDS in an Irish population from 1996 to 2000. METHOD: All reported cases of drug overdoses with AEDs from 1996 to 2000 were reviewed. Data was provided by the National Poisons Information Centre, Beaumont Hospital and the Central Statistics Office. Medical records from Beaumont Hospital were reviewed in specific cases of serious drug toxicities. An extensive review of published literature reviewing the safety profile of these AEDs was performed and medical literature retrieved from the databases of the pharmaceutical industry was similarly reviewed. RESULTS: Of the 164 reported cases of newer AEDs, there were no fatalities among the cases followed up. CONCLUSION: The absence of mortalities and serious consequences from deliberate self-poisoning with the newer agents is supportive evidence for the superior safety profile of the newer AEDs. PMID- 15797350 TI - Intravenous valproate as an innovative therapy in seizure emergency situations including status epilepticus--experience in 102 adult patients. AB - PURPOSE: The emergency treatment of seizures is an important practical issue, in particular the therapy of status epilepticus. Antiepileptic drugs for this condition should be easy to use, show rapid action, have a long-lasting antiepileptic effect, and have minimal cardiopulmonary and other side-effects. Unfortunately, none of the presently available medications such as phenytoin and barbiturates seems to have all of these four properties. Intravenous valproate became available some years ago and first experiences show promising safety data and efficacy results. METHODS: We report a series of 102 adult patients who received standardized high dosage intravenous valproate in various emergency situations, including status epilepticus. The therapeutic goal was persistent seizure control, defined as successful interruption of clinical seizure activity within less than 15 min, followed by seizure freedom during intravenous therapy for at least 12h. All side effects were documented. RESULTS: In 83/97 patients (85.6%) the therapeutic goal was achieved. Serious side effects were not documented in any patient. In particular there was no evidence of sedation, cardiorespiratory disturbances and hypotension as often seen in barbiturates and phenytoin. Mild side effects occurred in seven cases (6.9%). CONCLUSIONS: The intravenous application of VPA seems to be an easy-to-use, safe and efficient formulation as an alternative to phenytoin in all seizure emergency situations including status epilepticus. Further controlled comparison studies have to be performed in the future. PMID- 15797349 TI - Factors contributing to the stigma of epilepsy. AB - PURPOSE: To evaluate the factors, including personality and coping styles, likely to be influential in enhancing the social stigma of epilepsy. METHODS: Data were collected from 400 adults with epilepsy recruited from 10 epilepsy centers in Korea. Clinical information about seizures was obtained by neurologists, and other information was collected from self-completed questionnaires, including those measuring stigma scales. RESULTS: Thirty-one percent of people with epilepsy felt stigmatized by their condition and in 9% of these the stigma was severe. Multivariate analysis identified experiences of actual discrimination from society, introverted personality, problem solving controllability, and emotional subscale of QOLIE-31 as being independently associated with the social stigma of epilepsy. CONCLUSIONS: Episodes of discrimination, coping strategies, and personality may be important in feeling the stigma of epilepsy. These findings may provide a basis for further studies to clarify the causative factors generating the stigma of epilepsy. PMID- 15797351 TI - Antiepileptic drugs: a study of 1028 cases registered by the Sao Paulo Intoxication Control Center. AB - PURPOSE: To illustrate the profile of the cases of intoxications by antiepileptic drugs (AEDs) registered within a large urban area during the period of 1 year. METHODS: A cross-sectional observational study was performed in the Intoxication Control Center of the Sao Paulo City Hospital. We evaluated 6535 medical consults of intoxication cases involving prescription and over-the-counter drugs in 2001. We selected and analyzed the cases involving AEDs. Statistical analysis was performed in order to evaluate the frequency of different drug types and the circumstance of drug exposure. RESULTS: The most frequently observed AEDs were phenobarbital, carbamazepine, diazepam, and clonazepam. The frequency of intoxication cases among different age groups did not significantly differ according to the drug type. The circumstances involving AEDs intoxication were analogous to the circumstances of the cases of intoxication involving all other medications (p>0.05). CONCLUSIONS: There are a large number of intoxications involving AEDs. Imposing a restriction access to AEDs can be deleterious to patients that depend on AEDs, however, a strict program of AEDs distribution should be considered to reduce the cases of AED intoxication. PMID- 15797352 TI - Incidence of epilepsy and predictive factors of epileptic and non-epileptic seizures. AB - PURPOSE: To estimate the incidence of unprovoked seizures (US) and epilepsy in a general population from the southern part of the Netherlands, in relation to age, sex, etiology and seizure type, and to identify predictive factors of the epileptic and non-epileptic seizures. METHODS: All patients aged > or =14 years with a first seizure or who had undiagnosed seizures before the study period were included. Patients were identified from different sources and were independently evaluated and classified by a team of neurologists. A predictive profile for the occurrence of epileptic and non-epileptic seizures was obtained by stepwise logistic regression analysis. RESULTS: The overall annual incidence was 55/100,000 and 30/100,000 for US and epilepsy, respectively. The age-specific annual incidence of US and epilepsy increased with age and reached 120/100,000 and 62/100,000 for the > or =65 years of age group, respectively. The incidence of epilepsy and US in males was higher than in females and partial seizures prevailed over generalized seizures (40 versus 9/100,000). In up to 35% of the cases with US or epilepsy, the etiology was mainly cerebrovascular disease and brain tumors. Predictors for epileptic versus non-epileptic seizures of organic origin were an epileptiform EEG pattern (OR=0.06) versus a history of hypertension (OR=2.8) or cardiovascular disease (OR=5.4). Strong predictors for seizures of non-organic origin were female sex (OR=2.2) and head injury (OR=2.4). CONCLUSIONS: The incidence of US and epilepsy (overall, and age-, sex-, seizure specific) was similar to those reported by other developed countries. The predictive factors found in this study may assist in the early diagnosis of seizures. PMID- 15797353 TI - Efficacy and safety of topiramate in infants according to epilepsy syndromes. AB - Studies of the efficacy of topiramate (TPM) in infants and young children are few. Here we report an open, prospective, and pragmatic study of effectiveness of TPM in terms of epilepsy syndromes, in children aged less than 2 years. The median follow-up period was 11 months. We enrolled 59 children in the study: 22 affected by localization-related epilepsy (LRE), 23 by generalized epilepsy, six by Dravet's syndrome, and eight with unclassifiable epilepsy. TPM was effective (responders showed a decrease of more than 50% in seizure frequency) in 47% of patients, including 13% who were seizure-free at the last visit. TPM was more effective in localization-related epilepsy (48% of responders) than in generalized epilepsy (32% of responders). In the latter group, 19 patients suffered from infantile spasms. Four of six patients with cryptogenic infantile spasms became seizure-free. Of the 13 patients with symptomatic infantile spasms, only one was seizure-free. Results were poor for patients with Dravet's syndrome. In general, TPM was well tolerated. The most frequently reported adverse effects were drowsiness, irritability, hyperthermia, and anorexia. The present study concludes that TPM is effective for a broad range of seizures in infants and young children and represents a valid therapeutic option in this population. PMID- 15797354 TI - Behaviour problems in children and adolescents with seizure disorder: associations and risk factors. AB - PROBLEM: Fifty million people worldwide have epilepsy of whom 33 million are children. Children and adolescents with epilepsy are known to have high rates of behaviour problems and psychiatric co-morbidity. No studies on this topic have been conducted in developing countries, although 90% of such children live in the developing world. METHODS: The current study, set in Vellore in India, assessed the prevalence of psychopathology and its associations in children and adolescents with seizure disorder. One hundred and thirty two children were rated for psychopathology on the Child Behaviours Check List, and those who qualified as psychiatric cases were compared with those who did not. RESULTS: Seizure disorder was associated with significant psychopathology in 53.8% children. Belonging to a higher income group and living in an urban area (OR: 7.61, 95% CI: 2.78-20.8, p=0.0001), having longer than 3 years of illness (OR: 2.39, 95% CI: 2.18-5.67, p=0.03) and being treated by more than one anti-epileptic drug (OR: 3.08, 95% CI: 1.09-8.72, p=0.03) independently predicted psychopathology. CONCLUSIONS: Psychopathology reflected a complex interaction of seizures, socio demographic factors and treatment related variables, in accord with studies in industrialised countries. Clinicians should be sensitive to identifying psychopathology since this may lead to effective interventions for this group of children. PMID- 15797355 TI - Complaints associated with the use of antiepileptic drugs: results from a community-based study. AB - BACKGROUND: Few data exist with respect to the occurrence of chronic side effects due to antiepileptic drugs (AED) in routine clinical practice. OBJECTIVE: To evaluate the prevalence of subjective complaints which patients with epilepsy regard as side effects of their AED treatment in a community-based population. METHODS: Cross-sectional study. Subjects were identified through the database of AED-use in the pharmacies in a suburban area in The Netherlands. Respondents completed a brief questionnaire about their epilepsy, including a checklist with 30 complaints, which are common in AED users. RESULTS: We present data of 346 responding adults with treated epilepsy from a population of 107,000 adult inhabitants. Eighty percent was using monotherapy, with few patients taking new AEDs. Almost 60% of the patients reported complaints probably due to side effects in at least three domains. General CNS-related side effects were reported most often; memory problems (21.4% of the patients) and fatigue (20.3%) were dominant. Polytherapy was associated with more side effects than monotherapy. We identified differences in profiles of complaints between valproate, carbamazepine and phenytoin monotherapy. Complaints were not substantially associated with ongoing seizures or other treatment factors. CONCLUSIONS: The majority of patients taking AEDs for epilepsy think they have side effects form their drugs, even when seizures were in remission and when monotherapy was used. Our findings suggest a need to improve monitoring of complaints of side effects of AEDs and to explore the feasibility of interventions aimed at reduction of such complaints in everyday clinical practice. PMID- 15797356 TI - Educational problems with underlying neuropsychological impairment are common in children with Benign Epilepsy of Childhood with Centrotemporal Spikes (BECTS). AB - INTRODUCTION: Benign Epilepsy of Childhood with Centrotemporal Spikes (BECTS) is one of the most common childhood epilepsies with a good prognosis regarding the seizure and neuropsychological outcomes. However, recent reports indicate the presence of neuropsychological problems in a significant percentage of children with BECTS. Our study was aimed to examine the educational performance and neuropsychological functions along with clinical and electrographic characteristics in a cohort of children with BECTS. METHODS: We identified a cohort of children with BECTS by screening medical and EEG recordings of patients attending our institute. Data were collected with a standard protocol. Their educational performance was evaluated by an interview with the parents. Neuropsychological and language tests were administered to children who had educational problems. Statistical analysis was done using the chi2-test. RESULTS: Fifty children (29 boys and 21 girls; mean age of onset of epilepsy 7.84+/-2.87 years) who met the criteria for BECTS were included in this study. Atypical seizure characteristics for BECTS were observed in 26 (52%) children. EEG showed typical centrotemporal spike and wave discharges in all children, 42% of them had a tangential dipole in the frontocentral region. An additional extra-rolandic focus in the EEG was found in seven children (14%). Educational problems were identified in 27 children (54%); 19 of them had neuropsychological or language impairment (p=0.003). We found a statistically significant correlation between the occurrence of educational problems and the absence of a tangential dipole in the EEG (p<0.001). Abnormal language function had a significant correlation with atypical seizure semiology (p=0.021). CONCLUSION: This study shows that a significant number of children with BECTS have neuropsychological impairment and educational problems. PMID- 15797357 TI - Diagnostic performance of 18F-FDG PET and ictal 99mTc-HMPAO SPET in pediatric temporal lobe epilepsy: quantitative analysis by statistical parametric mapping, statistical probabilistic anatomical map, and subtraction ictal SPET. AB - We investigated the diagnostic performance of 18F-FDG PET and ictal (99m)Tc-HMPAO SPET in pediatric temporal lobe epilepsy (TLE). Twenty-one pediatric TLE patients were enrolled in this study. Their diagnoses were confirmed by histology and post surgical outcome (Engel class I or II). The patients' ages were 18 or younger (15+/-3 years). Of the 21 patients, 21 patients underwent 18F-FDG PET scan and 15 underwent ictal (99m)Tc-HMPAO SPET. Preoperative PET and/or ictal SPET images were reviewed by simple visual assessment and by statistical parametric mapping (SPM). Asymmetric indices (AI) were calculated using statistical probabilistic anatomical map (SPAM) on 18F-FDG PET. In nine patients who underwent both ictal and interictal SPET, SISCOM (subtraction ictal SPET coregistered to MR template) was performed. PET correctly localized epileptogenic zones in 20 of 21 (95%) by visual assessment. SPM analysis of PET correctly localized epileptogenic zones in 18 of 21 (86%). Ictal SPET correctly localized epileptogenic zones in 12 of 15 (80%) by visual assessment. SPM analysis of ictal SPET correctly localized epileptogenic zones in 12 of 15 (80%). SISCOM correctly localized 8 of 9 (89%), which was equal to that of visual assessment of ictal SPET. The AIs of the temporal lobes by PET were -15+/-8.4 in the left and 9.9+/-8.9 in the right TLE (normal control: -2.9+/-2.8), and correctly localized epileptogenic zones in all cases. As is found in adult TLE, PET and ictal SPET efficiently localized epileptogenic zones in pediatric TLE. SPM analysis of PET or ictal SPET could be used as an aid to visual assessment. Moreover, SISCOM was equal visual assessment of ictal SPET images in terms of lesion localizations. PMID- 15797358 TI - Predominance of rotavirus G9 genotype in children hospitalized for rotavirus gastroenteritis in Belgium during 1999-2003. AB - BACKGROUND: Group A rotavirus genotypes G1, G2, G3 and G4 are the main etiological agents of infantile diarrhea. The G9 rotavirus has recently emerged as a fifth important genotype all over the world. OBJECTIVE: To characterize the VP7 gene of group A rotaviruses from gastroenteritis patients admitted to the Gasthuisberg University Hospital, Leuven, Belgium, during 1999-2003. STUDY DESIGN: Rotavirus antigen was detected in stool specimens using an enzyme immunoassay. G-typing was performed by reverse transcriptase polymerase chain reaction (RT-PCR) amplification and sequencing of the complete VP7 gene. RESULTS: The genotype distribution varied markedly over the four rotavirus years in Belgium. In the 1999-2000 rotavirus year, G1 was the predominating type (72%), and G9 was present in 5% of the rotavirus-positive patients. In the 2000-2001 and 2002-2003 years, G9 appeared as the dominating strain (45% and 53%, respectively). In the 2001-2002 year, between two G9 epidemic years, G1 was dominating (66%) but G9 was still present in 24%. All the G9 isolates were combined with P[8] and shared a high gene sequence similarity (<3% sequence divergence on the nucleotide and amino acid level). Phylogenetic analysis of the VP7 genes revealed that our Belgian G9 strains clustered together with recent G9 strains from all over the world, distinct from the prototype G9 strains isolated in the 1980s. CONCLUSION: Our study indicates that although the first introduction of G9 isolates in the Belgian population was recorded in 1997, G9 strains were able to establish themselves quickly as the predominant genotype. The emergence of G9 as an important pathogen in both developing as industrialized countries necessitates the urgent consideration of the G9 moiety in rotavirus vaccines. PMID- 15797359 TI - The emergence of rotavirus genotype G9 in hospitalised children in Slovenia. AB - BACKGROUND: Rotavirus G9 genotype was thought to be the fifth most common genotype circulating amongst the population. In previous studies in Slovenia, only G1, G3 and G4 genotypes were detected. OBJECTIVES: To determine G and P genotypes of rotaviruses causing dehydrating gastroenteritis in children hospitalised at the University Medical Centre Ljubljana during the winter season 2001-2002. Some data obtained in previous years are included, too. STUDY DESIGN: For the G and P genotypes determination, we selected 99 of the total of 565 rotavirus positive samples. RT-PCR was carried out for G gene or partial P gene amplification. The RT-PCR product was used as a template for multiplex nested PCR using genotype-specific primers. In untypable samples, a sequence analysis of a short segment of G or P gene was performed. From the period before July 2001, 183 stool samples were examined using the same methods. RESULTS: Genotype G1 was determined in 37, G4 in 6, and G9 in 28 samples out of 99. Only one sample showed a mixed infection with G1G4 genotype specifics. Following the sequence analysis of the short segment of G gene in 11 G9 genotypes, 2 different clusters of G9 genotype were determined. All samples had the same P genotype--P[8]. G9 genotype had not been detected prior to July 2001. CONCLUSION: Rotavirus G9 genotype emerged in Slovenia in the year 2001. Two different clusters were determined which have to be further characterised in detail. PMID- 15797360 TI - Development and evaluation of an enzyme-linked immunosorbent assay for detection of antibodies against the spike protein of SARS-coronavirus. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) is caused by infection with SARS-associated coronavirus (CoV). Amino acid residues 450-650 of the spike (S) glycoprotein of SARS-CoV (S450-650) contains dominant epitopes for anti-viral antibodies (Abs) in patient sera. OBJECTIVES: To develop and evaluate an ELISA system for detection of anti-S Abs in patient sera. STUDY DESIGN: Express recombinant S450-650 in E. Coli and evaluate the sensitivity and specificity of an ELISA system based on the S450-650 polypeptide. RESULTS: The S450-650-based ELISA detected IgG Abs in 41 out of 51 serum samples from 22 hospitalized patients with probable SARS, a result closely correlated with that obtained with a virus-based ELISA (r = 0.75, k = 0.8). Differential anti-S IgG responses were observed amongst SARS patients. Some of them produced anti-S Abs early during their infection, while others failed to make IgG Abs against the S450-650 polypeptide. None of the serum samples from 100 healthy blood donors was positive in the S450-650-based assay. CONCLUSION: The S450-650-based ELISA can detect anti S IgG Abs with high sensitivity and specificity. PMID- 15797361 TI - Clinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods. AB - BACKGROUND: The protocols of WHO network laboratories facilitated development of rapid diagnosis for SARS coronavirus (CoV) using reverse transcription (RT)-PCR assays. However, several reports have shown that conventional and real-time PCR assays were very specific for SARS CoV but lack sensitivity depending on the assay, specimen, and time course of disease. OBJECTIVE: To evaluate an automatic nucleic acid extraction system and two standardized real-time PCR assays for rapid diagnosis of SARS CoV during outbreak and post-epidemic periods in Hong Kong. STUDY DESIGN: Specimens from clinically suspected SARS patients collected during outbreak and post-epidemic periods were tested by an automatic nucleic acid extraction system followed by our first generation conventional RT-PCR and two standardized real-time PCR assays (Artus GmbH, Germany and Roche Diagnostics, Germany). Paired serum samples were assayed for increasing titer against SARS CoV. RESULTS: In the SARS epidemic, Artus and Roche PCR assays exhibited sensitivities of 87% and 85% for respiratory specimens (n = 64), 91% and 88% for stool (n = 44), and 82% for urine (n = 29). A specificity of 100% was exhibited by both PCR assays except Artus attained only a 92% specificity for stool. For post-epidemic period, no SARS CoV was identified among 56 respiratory specimens by all PCR assays. Inhibitors to PCR assays were detected at an average rate of 7 8% among 202 clinical specimens. CONCLUSION: This study highlights the high throughput and performance of automatic RNA extraction in coordination with standardized real-time PCR assays suitable for large-scale routine diagnosis in case of future SARS epidemic. As no SARS CoV was detected among specimens collected during post-epidemic period, the positive predictive value of real-time PCR assays for detection of SARS CoV during low epidemic requires further evaluation. PMID- 15797363 TI - Follow-up of 28 HCMV seropositive renal-transplant recipients: comparison of clinical, biological and virological parameters in the groups of treated versus untreated infected patients. AB - BACKGROUND: A pre-emptive strategy for prevention of Human Cytomegalovirus (HCMV) disease depends on accurate detection of HCMV infection and clinical and/or biological abnormalities. OBJECTIVES: The aim of our study was to evaluate a therapeutic strategy based on the presence of either minimal clinical and/or biological symptoms or high HCMV viral load assessed by quantitative real-time PCR from whole blood as previously described. STUDY DESIGN: Between June 2002 and July 2003, 70 HCMV seropositive patients underwent a renal transplantation. Virological monitoring was performed every 2 weeks until day 90 then every 3-4 weeks until day 180. Biochemical and haematological parameters were also prospectively monitored. RESULTS: Twenty-eight patients (40%) showed at least one positive HCMV DNA in whole blood. Based on the following criteria: HCMV viral load greater than 4log(10)/ml, or the persistence of a HCMV DNAemia in two consecutive blood samples, or fever, or leucopenia or neutropenia, or increase in alanine aminotransferase level, 14 of the 28 patients received IV ganciclovir as a pre-emptive treatment. Immunosuppressive therapy and demographical data were comparable in both groups. As far as virological, haematological and biochemical parameters are concerned, no statistical significant difference was observed between treated and untreated patients. Moreover no adverse outcome was observed among untreated patients always having a HCMV viral load below 4log(10)/ml. CONCLUSIONS: This study reinforces the need of monitoring of HCMV seropositive renal-transplant recipients based on HCMV diagnosis and quantification by real time PCR. The results showed that HCMV viraemic patients may benefit of absence of antiviral treatment when they have a low viral load (below 4log(10)/ml) and absence of clinical and/or biological abnormalities. Further studies are required now to validate the threshold value at which we should begin pre-emptive therapy. PMID- 15797362 TI - Simultaneous detection of herpes simplex virus types 1 and 2 by real-time PCR and Pyrosequencing. AB - BACKGROUND: Up to 80% of the US adult population has been exposed to herpes simplex virus (HSV) type 1, primarily during childhood. Also, approximately 20% of the US population has contracted genital herpes from HSV-2 infections. Clinical symptoms can present as fever, headache, malaise, myalgia, and cold sores/lesions that cause pain, itching, dysuria, and vaginal or urethral discharge. A recurrence of infection is common. HSV culturing is characterized by low sensitivity with variable success rates due to shipping conditions. OBJECTIVE: To design and validate a real-time PCR assay capable of simultaneously detecting each HSV subtype. STUDY DESIGN: ATCC-purchased HSV-1 and HSV-2 positive samples and HSV-1 and HSV-2 infected clinical specimens were assayed simultaneously with shared amplification primers and subtype-specific probes against the HSV glycoprotein B gene on a Rotor-Gene 3000 platform. Separately, two PCR reactions were performed in which one primer contained a 5' biotin modification. Single-stranded DNA from the amplicon was purified and Pyrosequenced. RESULTS: The quantitative range of the assay extended from 10(8) through 10(0) copies of each virus (r(2) > 0.991) and specificity was determined by non-amplification of 37 different human pathogens, including other herpesviruses such as VZV, CMV, and EBV. Sensitivity and specificity values of 100% were calculated by concordance analysis between the real-time PCR and the DNA Pyrosequencing results (HSV-1: n = 119, HSV-2: n = 120). Application of this assay to 4581 cervical swab specimens collected from women visiting physicians primarily in six states provided detection rates of 3.1% for HSV-1 and 7.6% for HSV-2. The average age of women infected with HSV-1 was 29.5 versus 35.6 for HSV 2. CONCLUSIONS: This procedure was demonstrated as both highly sensitive and specific for the detection of HSV-1 and HSV-2 in a single reaction. Also, the integration of Pyrosequencing analysis permitted an innovative and rapid verification for each subtype. PMID- 15797364 TI - Comparative evaluation of the Cobas Amplicor HIV-1 Monitor Ultrasensitive Test, the new Cobas AmpliPrep/Cobas Amplicor HIV-1 Monitor Ultrasensitive Test and the Versant HIV RNA 3.0 assays for quantitation of HIV-1 RNA in plasma samples. AB - BACKGROUND: There are several commercially available assays for the quantitation of HIV RNA. A new automated specimen preparation system, the Cobas AmpliPrep, was developed to automate this last part of the PCR. OBJECTIVES AND STUDY DESIGN: We compared the results obtained by the Roche Cobas Amplicor HIV-1 Monitor Ultrasensitive Test (MCA, manual sample preparation) with those by the Versant HIV-1 RNA 3.0 assay (bDNA). Secondly we compared the MCA with the new Cobas AmpliPrep/Cobas Amplicor HIV Monitor Ultrasensitive Test (CAP/CA, automated specimen preparation) by investigating clinical patient samples and a panel of HIV-1 non-B subtypes. Furthermore, we assessed the assay throughput and workflow (especially hands-on time) for all three assays. RESULTS: Seventy-two percent of the 140 investigated patient samples gave concordant results in the bDNA and MCA assays. The MCA values were regularly higher than the bDNA values. One sample was detected only by the MCA within the linear range of quantification. In contrast, 38 samples with results <50 copies/ml in the MCA showed in the bDNA results between 51 and 1644 copies/ml (mean value 74 copies/ml); 21 of these specimens were shown to have detectable HIV RNA < 50 copies/ml in the MCA assay. The overall agreement between the MCA and the CAP/CA was 94.3% (551/584). The quantification results showed significant correlation, although the CAP/CA generated values slightly lower than those generated by the manual procedure. We found that the CAP/CA produced comparable results with the MCA test in a panel of HIV-1 non-B subtypes. CONCLUSIONS: All three assays showed comparable results. The bDNA provides a high sample throughput without the need of full automation. The new CAP/CA provides reliable test results with no HIV-subtype specific influence and releases time for other works in the laboratory; thus it is suitable for routine diagnostic PCR. PMID- 15797365 TI - Molecular characterization of Kaposi's sarcoma associated herpesvirus (KSHV) from patients with AIDS-associated Kaposi's sarcoma in Sao Paulo, Brazil. AB - BACKGROUND: Kaposi's sarcoma (KS) is caused by Kaposi's sarcoma associated herpesvirus (KSHV/HHV-8), the eighth Herpesvirus found to infect humans. The molecular epidemiology of KSHV is related closely to ethnicity and geographical location of studied populations. There is little epidemiological and molecular information about KSHV strains circulating in Brazil. OBJECTIVES: To characterize KSHV strains isolated from AIDS patients with Kaposi's sarcoma (AIDS-KS) in Sao Paulo, Brazil, and to examine associations between KSHV subtypes, ethnicity and HIV risk categories. METHODS: AIDS-KS patients were recruited consecutively at the largest AIDS reference hospital in Sao Paulo. Fragments (420 bp) of the VR1 and VR2 regions of KSHV open reading frame (ORF) K1 were amplified by nested PCR and sequenced directly. RESULTS: We analysed 37 samples from 33 patients, and found subtypes A-C in 48%, 21% and 30% of patients respectively, including two patients infected with subtype A5, a first report from Brazil. Sexual orientation was associated with subtype: 12/14 (86%) patients with subtype A were male homo/bisexual, compared with 3/8 (38%) among patients infected with subtype C (P = 0.05). A higher proportion of male patients with subtype C were of Caucasian origin (7/8 (87%)), compared with 7/16 (44%) among male patients with subtype A (P = 0.08). CONCLUSIONS: This first detailed report of KSHV subtypes among AIDS KS patients in Brazil reports the first isolation of KSHV subtype A5 in this country, and suggests KSHV strain transmission between different ethnic groups, and association of specific strains with sexual orientation. PMID- 15797366 TI - Mitochondrial DNA depletion in adipose tissue of HIV-infected patients with peripheral lipoatrophy. AB - BACKGROUND: NRTI-induced host toxicity is proposed to involve cellular mitochondrial DNA (mtDNA) depletion. Determinants of cellular mtDNA copy number from HIV-infected patients receiving HAART and HIV-seronegative controls were investigated from subcutaneous fat samples, and relation with antiretroviral regimen was studied. STUDY DESIGN: HIV-infected patients receiving HAART (n = 50), HIV-infected patients not currently under HAART regimen (n = 2) and HIV seronegative controls (n = 9) of similar age and BMI were enrolled prospectively when undergoing Coleman's lipostructure for correction of facial lipoatrophy or plastic surgery, respectively. After centrifugation, abdominal fat tissue was collected and stored at -80 degrees C. MtDNA analysis was blindly performed after a total DNA extraction from adipose tissue, followed by a real-time PCR quantification. The log of mtDNA copies/cell in adipose tissue [log(DNA)] was compared between groups by means of analysis of variance. RESULTS: The log(DNA) in adipose tissue of HIV-infected patients was significantly lower than in the HIV-seronegative control group (P < 0.0001). In HIV-infected patients, log(DNA) was significantly reduced in the 50 NRTI-treated patients (P < 0.01), but not when considering mtDNA level according to the use of PI or NNRTI in current HAART regimen. In NRTI-treated patients, only stavudine (n = 20) and didanosine (n=14) were significantly and independently associated with reduced mtDNA level (P < 0.0001 and <0.05, respectively). Currently stavudine or didanosine-treated patients had a significant reduced mtDNA level compared to past users (P < 0.0001 and <0.05, respectively). Other clinical, biological, and immuno-virological variables than NRTI did not correlate significantly to adipocyte mtDNA level. CONCLUSION: This study supports that current treatment by NRTI is a main determinant of mtDNA depletion in adipose tissue of HIV-seropositive patients with peripheral fat wasting. Stavudine or didanosine current intake is significantly associated with mtDNA depletion in vivo, that could be reversible after the discontinuation of these molecules, when considering mtDNA level according to current use versus past use of these molecules. PMID- 15797367 TI - Molecular epidemiology of HTLV-2 infection among intravenous drug users in Spain. AB - BACKGROUND: HTLV-2 infection is prevalent among intravenous drug users (IDUs), which often are coinfected with HIV-1. Two main subtypes, HTLV-2a and -2b, have been described among European IDUs, with a characteristically geographic distribution: IIa in northern Europe and IIb in the south. Although frequent traveling within Europe might have favoured a wide dissemination of diverse HTLV 2 variants, HTLV-2b was so far the only subtype reported in Spain. OBJECTIVE: To investigate the current molecular epidemiology of HTLV-2 in Spain. STUDY DESIGN: Twenty-six new HTLV-2 strains isolated from IDUs over the last 11 years in different geographic regions were examined. The HTLV LTR region (620bp) was sequenced and phylogenetic analyses were performed. Endonuclease restriction sites were examined to further characterize the HTLV-2 subgroup. RESULTS: All 26 individuals infected with HTLV-2 showed viruses belonging to the HTLV-2 b4 clade and were closely related to the previously reported HTLV-2 Spanish and Italian IDU isolates. CONCLUSIONS: HTLV-2 subtype b4 continues to be the only HTLV-2 subgroup recognized so far in Spain and no introduction of other HTLV-2 variants has occurred over the last 11 years. PMID- 15797368 TI - High frequency of parvovirus B19 DNA in bone marrow samples from rheumatic patients. AB - BACKGROUND: Human parvovirus B19 (B19) polymerase chain reaction (PCR) is now a routine analysis and serves as a diagnostic marker as well as a complement or alternative to B19 serology. The clinical significance of a positive B19 DNA finding is however dependent on the type of tissue or body fluid analysed and of the immune status of the patient. OBJECTIVES: To analyse the clinical significance of B19 DNA positivity in bone marrow samples from rheumatic patients. STUDY DESIGN: Parvovirus B19 DNA was analysed in paired bone marrow and serum samples by nested PCR technique. Serum was also analysed for B19-specific IgG and IgM antibodies and the results were compared with clinical and epidemiological data. RESULTS AND CONCLUSIONS: B19 IgG was found in 41 of 50 patients (82%) whereas none was B19 IgM positive. The serologic evaluation showed that none of the patients had acute B19 infection. However, B19 DNA was detected by PCR in 13 of 50 (26%) bone marrow samples from these patients indicating a high frequency of persistent infection compared with previous reports of patient groups and healthy controls. In the study, 22 patients had rheumatoid arthritis (RA) and 7 of these RA patients were B19 DNA positive in bone marrow. Rheumatoid factor was positive in 4 of the 7 B19 DNA positive RA patients as compared with Rheumatoid factor positivity in all of the 15 B19 DNA negative RA patients. Erosive arthritis in X-ray was less common in the B19 DNA positive group than in the B19 DNA negative group. A high frequency of parvovirus B19 DNA was thus detected in bone marrow samples in rheumatic patients. The clinical data does not support a direct association between B19 PCR positivity and rheumatic disease manifestation. Therefore, the clinical significance of B19 DNA positivity in bone marrow samples from rheumatic patients must be interpreted with caution. PMID- 15797369 TI - Intermittent viremia during first-line, protease inhibitors-containing therapy: significance and relationship with drug resistance. AB - BACKGROUND: In HIV-infected patients on first-line antiretroviral therapy, the significance of intermittent viremia and their relationship with drug resistance remain unclear. OBJECTIVE: To study the virological characteristics of intermittent viremia (IV) and the association between IV and later virological failure (VF) in patients on a first-line, PI-containing therapy. STUDY DESIGN: Antiretroviral-naive patients were enrolled in the APROVIR substudy of the prospective, multicenter APROCO cohort at the time they initiated a PI-containing therapy and were followed-up at month 1 and every 2 months. IV was defined as plasma HIV-1 RNA > 500 copies/ml on a single specimen. VF were defined as: (1) viral rebound on two consecutive plasma specimens with HIV-1 RNA > 500 copies/ml after an initial response below 500 copies/ml, or (2) persistence of plasma HIV-1 RNA> or =500 copies/ml during the first year of follow-up. Genotypic resistance analysis was performed at baseline and at the time of IV. PI plasma concentrations were determined at the time of IV. RESULTS: IV was found in 20/219 patients in a 2 years follow-up. The occurrence of IV in the first year of therapy was associated with a higher risk of virological failure during the second year (p = 0.03). Genotypic resistance at the time of IV was found in only 4/16 patients and was not predictive of a subsequent virological failure. PI plasma levels suggested lack of adherence in 50% of patients with IV. CONCLUSION: The occurrence of IV > 500 copies/ml among patients on first-line, PI-containing ART is suggestive of a lack of adherence rather than the selection of resistant variants and should lead to an intensification of adherence monitoring in order to reduce the risk of subsequent VF. PMID- 15797370 TI - I wish to comment on several points in the recent "Guidelines". PMID- 15797371 TI - Clinical outcome of long-term lamivudine treated chronic HBV infected and HIV/HBV coinfected patients. PMID- 15797372 TI - The molecular biography of the cell. PMID- 15797373 TI - An unexpected role for ubiquitylation of a transcriptional activator. AB - The yeast transcriptional activator Gal4 has served as a paradigm for understanding how eukaryotic cells mount rapid transcriptional responses to environmental changes. In this issue of Cell, Muratani et al. (2005) provide evidence that Gal4 ubiquitylation and destruction are required for activation by Gal4. Surprisingly, this modification is required at a postinitiation step in transcription for the production of mRNAs that are correctly processed and fully functional for translation. PMID- 15797374 TI - San1p, checking up on nuclear proteins. AB - Cellular quality control mechanisms perform vital tasks by ensuring that the proteome reflects precisely the information encoded by the genome. In this issue of Cell, Gardner et al. (2005) report the discovery of a novel protein quality control system that resides in the nucleus. Central to this system is the E3 ligase San1p, which monitors nuclear proteins and targets aberrant species for destruction. PMID- 15797375 TI - Axis formation--beta-catenin catches a Wnt. AB - In this issue of Cell, the Heasman group implicates Wnt11 as a component of the canonical Wnt signaling pathway that specifies Xenopus laevis axis formation (Tao et al., 2005). This important work not only identifies a long-sought-after dorsalizing factor but also highlights the pivotal role of extracellular cofactors in specifying the activation of either canonical or noncanonical Wnt pathways. PMID- 15797376 TI - How do so few control so many? AB - The separation of sister chromatids at the metaphase-to-anaphase transition is triggered by a protease called separase that is activated by the destruction of an inhibitory chaperone (securin). This process is mediated by a ubiquitin protein ligase called the anaphase-promoting complex or cyclosome (APC/C), along with a protein called Cdc20. It is vital that separase not be activated before every single chromosome has been aligned on the mitotic spindle. Kinetochores that have not yet attached to microtubules catalyze the sequestration of Cdc20 by an inhibitor called Mad2. Recent experiments shed important insight into how Mad2 molecules bound to centromeres through their association with a protein called Mad1 might be transferred to Cdc20 and thereby inhibit securin's destruction. PMID- 15797377 TI - The mTOR inhibitor RAD001 sensitizes tumor cells to DNA-damaged induced apoptosis through inhibition of p21 translation. AB - Although DNA damaging agents have revolutionized chemotherapy against solid tumors, a narrow therapeutic window combined with severe side effects has limited their broader use. Here we show that RAD001 (everolimus), a rapamycin derivative, dramatically enhances cisplatin-induced apoptosis in wild-type p53, but not mutant p53 tumor cells. The use of isogenic tumor cell lines expressing either wild-type mTOR cDNA or a mutant that does not bind RAD001 demonstrates that the effects of RAD001 are through inhibition of mTOR function. We further show that RAD001 sensitizes cells to cisplatin by inhibiting p53-induced p21 expression. Unexpectedly, this effect is attributed to a small but significant inhibition of p21 translation combined with its short half-life. These findings provide the molecular rationale for combining DNA damaging agents with RAD001, showing that a general effect on a major anabolic process may dramatically enhance the efficacy of an established drug protocol in the treatment of cancer patients with solid tumors. PMID- 15797378 TI - The birnavirus crystal structure reveals structural relationships among icosahedral viruses. AB - Double-stranded RNA virions are transcriptionally competent icosahedral particles that must translocate across a lipid bilayer to function within the cytoplasm of the target cell. Birnaviruses are unique among dsRNA viruses as they have a single T = 13 icosahedral shell, lacking the characteristic inner capsid observed in the others. We determined the crystal structures of the T = 1 subviral particle (260 angstroms in diameter) and of the T = 13 intact virus particle (700 angstroms in diameter) of an avian birnavirus to 3 angstroms and 7 angstroms resolution, respectively. Our results show that VP2, the only component of the virus icosahedral capsid, is homologous both to the capsid protein of positive strand RNA viruses, like the T = 3 nodaviruses, and to the T = 13 capsid protein of members of the Reoviridae family of dsRNA viruses. Together, these results provide important insights into the multiple functions of the birnavirus capsid and reveal unexpected structural relationships among icosahedral viruses. PMID- 15797379 TI - The yeast APC/C subunit Mnd2 prevents premature sister chromatid separation triggered by the meiosis-specific APC/C-Ama1. AB - Cohesion established between sister chromatids during pre-meiotic DNA replication mediates two rounds of chromosome segregation. The first division is preceded by an extended prophase wherein homologous chromosomes undergo recombination. The persistence of cohesion during prophase is essential for recombination and both meiotic divisions. Here we show that Mnd2, a subunit of the anaphase-promoting complex (APC/C) from budding yeast, is essential to prevent premature destruction of cohesion in meiosis. During S- and prophase, Mnd2 prevents activation of the APC/C by a meiosis-specific activator called Ama1. In cells lacking Mnd2 the APC/C-Ama1 enzyme triggers degradation of Pds1, which causes premature sister chromatid separation due to unrestrained separase activity. In vitro, Mnd2 inhibits ubiquitination of Pds1 by APC/C-Ama1 but not by other APC/C holo enzymes. We conclude that chromosome segregation in meiosis depends on the selective inhibition of a meiosis-specific form of the APC/C. PMID- 15797380 TI - Mnd2, an essential antagonist of the anaphase-promoting complex during meiotic prophase. AB - Meiotic cohesin serves in sister chromatid linkage and DNA repair until its subunit Rec8 is cleaved by separase. Separase is activated when its inhibitor, securin, is polyubiquitinated by the Cdc20 regulated anaphase-promoting complex (APC(Cdc20)) and consequently degraded. Differently regulated APCs (APC(Cdh1), APC(Ama1)) have not been implicated in securin degradation at meiosis I. We show that Mnd2, a factor known to associate with APC components, prevents premature securin degradation in meiosis by APC(Ama1). mnd2Delta cells lack linear chromosome axes and exhibit precocious sister chromatid separation, but deletion of AMA1 suppresses these defects. Besides securin, Sgo1, a protein essential for protection of centromeric cohesion during anaphase I, is also destabilized in mnd2delta cells. Mnd2's disappearance prior to anaphase II may activate APC(Ama1). Human oocytes may spend many years in meiotic prophase before maturation. Inhibitors of meiotic APC variants could prevent loss of chiasmata also in these cells, thereby guarding against aberrant chromosome segregation. PMID- 15797381 TI - Degradation-mediated protein quality control in the nucleus. AB - Protein quality control degradation systems rid the cell of aberrant proteins, preventing detrimental effects on normal cellular function. Although such systems have been identified in most subcellular compartments, none have been found in the nucleus. Here, we report the discovery of such a system in Saccharomyces cerevisiae. It is defined by San1p, a ubiquitin-protein ligase that, in conjunction with the ubiquitin-conjugating enzymes Cdc34p and Ubc1p, targets four distinct mutant nuclear proteins for ubiquitination and destruction by the proteasome. San1p has exquisite specificity for aberrant proteins and does not target the wild-type versions of its mutant substrates. San1p is nuclear localized and requires nuclear localization for function. Loss of SAN1 results in a chronic stress response, underscoring its role of protein quality control in the cell. We propose that San1p-mediated degradation acts as the last line of proteolytic defense against the deleterious accumulation of aberrant proteins in the nucleus and that analogous systems exist in other eukaryotes. PMID- 15797382 TI - Mitochondrial presequence translocase: switching between TOM tethering and motor recruitment involves Tim21 and Tim17. AB - The presequence translocase of the inner mitochondrial membrane (TIM23 complex) operates at a central junction of protein import. It accepts preproteins from the outer membrane TOM complex and directs them to inner membrane insertion or, in cooperation with the presequence translocase-associated motor (PAM), to the matrix. Little is known of how the TIM23 complex coordinates these tasks. We have identified Tim21 (YGR033c) that interacts with the TOM complex. Tim21 is specific for a TIM23 form that cooperates with TOM and promotes inner membrane insertion. Protein translocation into the matrix requires a switch to a Tim21-free, PAM bound presequence translocase. Tim17 is crucial for the switch by performing two separable functions: promotion of inner membrane insertion and binding of Pam18 to form the functional TIM-PAM complex. Thus, the presequence translocase is not a static complex but switches between TOM tethering and PAM binding in a reaction cycle involving Tim21 and Tim17. PMID- 15797383 TI - SREBP pathway responds to sterols and functions as an oxygen sensor in fission yeast. AB - Cholesterol and fatty acid synthesis in mammals are controlled by SREBPs, a family of membrane bound transcription factors. Our studies identified homologs of SREBP, its binding partner SCAP, and the ER retention protein Insig in Schizosaccharomyces pombe, named sre1+, scp1+, and ins1+. Like SREBP, Sre1 is cleaved and activated in response to sterol depletion in a Scp1-dependent manner. Microarray analysis revealed that Sre1 activates sterol biosynthetic enzymes as in mammals, and, surprisingly, Sre1 also stimulates transcription of genes required for adaptation to hypoxia. Furthermore, Sre1 rapidly activates these target genes in response to low oxygen and is itself required for anaerobic growth. Based on these findings, we propose and test a model in which Sre1 and Scp1 monitor oxygen-dependent sterol synthesis as an indirect measure of oxygen supply and mediate a hypoxic response in fission yeast. PMID- 15797384 TI - Perp is a p63-regulated gene essential for epithelial integrity. AB - p63 is a master regulator of stratified epithelial development that is both necessary and sufficient for specifying this multifaceted program. We show here that Perp, a tetraspan membrane protein originally identified as an apoptosis associated target of the p53 tumor suppressor, is the first direct target of p63 clearly involved in mediating this developmental program in vivo. During embryogenesis, Perp is expressed in an epithelial pattern, and its expression depends on p63. Perp-/- mice die postnatally, with dramatic blistering in stratified epithelia symptomatic of compromised adhesion. Perp localizes specifically to desmosomes, adhesion junctions important for tissue integrity, and numerous structural defects in desmosomes are observed in Perp-deficient skin, suggesting a role for Perp in promoting the stable assembly of desmosomal adhesive complexes. These findings demonstrate that Perp is a key effector in the p63 developmental program, playing an essential role in an adhesion subprogram central to epithelial integrity and homeostasis. PMID- 15797385 TI - Maternal wnt11 activates the canonical wnt signaling pathway required for axis formation in Xenopus embryos. AB - Wnt signaling pathways play essential roles in patterning and proliferation of embryonic and adult tissues. In many organisms, this signaling pathway directs axis formation. Although the importance of intracellular components of the pathway, including beta-catenin and Tcf3, has been established, the mechanism of their activation is uncertain. In Xenopus, the initiating signal that localizes beta-catenin to dorsal nuclei has been suggested to be intracellular and Wnt independent. Here, we provide three lines of evidence that the pathway specifying the dorsal axis is activated extracellularly in Xenopus embryos. First, we identify Wnt11 as the initiating signal. Second, we show that activation requires the glycosyl transferase X.EXT1. Third, we find that the EGF-CFC protein, FRL1, is also essential and interacts with Wnt11 to activate canonical Wnt signaling. PMID- 15797386 TI - Facilitated transport of a Dpp/Scw heterodimer by Sog/Tsg leads to robust patterning of the Drosophila blastoderm embryo. AB - Patterning the dorsal surface of the Drosophila blastoderm embryo requires Decapentaplegic (Dpp) and Screw (Scw), two BMP family members. Signaling by these ligands is regulated at the extracellular level by the BMP binding proteins Sog and Tsg. We demonstrate that Tsg and Sog play essential roles in transporting Dpp to the dorsal-most cells. Furthermore, we provide biochemical and genetic evidence that a heterodimer of Dpp and Scw, but not the Dpp homodimer, is the primary transported ligand and that the heterodimer signals synergistically through the two type I BMP receptors Tkv and Sax. We propose that the use of broadly distributed Dpp homodimers and spatially restricted Dpp/Scw heterodimers produces the biphasic signal that is responsible for specifying the two dorsal tissue types. Finally, we demonstrate mathematically that heterodimer levels can be less sensitive to changes in gene dosage than homodimers, thereby providing further selective advantage for using heterodimers as morphogens. PMID- 15797388 TI - Adsorption at the liquid/liquid interface in mixed systems with hydrophobic extractants and modifiers 1. Study of equilibrium interfacial tension at the hydrocarbon/water interface in binary mixed systems. AB - Equilibrium interfacial tension at the liquid/liquid interfaces for two chelating metal ion extractants, 2-hydroxy-5-nonylacetophenone oxime (HNAF) and 1 phenyldecane-1,3-dion (beta-diketone), two solvating extractants, trioctylphosphine oxide (TOPO) and tributyl phosphate (TBP), and a modifier, decanol, were obtained with a drop volume tensiometer. Moreover, four equimolar binary mixtures of extractant/extractant and extractant/modifier type were considered. The composition of the mixed adsorbed monolayer and the molecular interaction parameters beta were determined by the Rosen equation. It was found that in all the studied systems coadsorption exists; however, synergism in the reduction of interfacial tension was not observed. The obtained results indicate that in the case of three mixtures considered the composition of a mixed monolayer at the hydrocarbon/water interface was quite different from that in the bulk organic phase. Only for the TOPO/beta-diketone mixture were the compositions at the interface and in the bulk organic phase similar. The obtained results indicate that it is impossible to predict the composition of a mixed monolayer by taking into account the interfacial activity of individual components of the mixture. In some cases the compound shows lower interfacial activity (smaller efficiency and effectiveness of adsorption) and occupies a dominant position at the interface, regardless of the type of hydrocarbon used as the organic diluent. PMID- 15797389 TI - Physicochemical characterization of the retardation of aqueous Cs+ ions by natural kaolinite and clinoptilolite minerals. AB - The aim of this study was to carry out kinetic, thermodynamic, and surface characterization of the sorption of Cs+ ions on natural minerals of kaolinite and clinoptilolite. The results showed that sorption followed pseudo-second-order kinetics. The activation energies were 9.5 and 13.9 kJ/mol for Cs+ sorption on kaolinite and clinoptilolite, respectively. Experiments performed at four different initial concentrations of the ion revealed that the percentage sorption of Cs+ on clinoptilolite ranged from 90 to 95, compared to 28 to 40 for the kaolinite case. At the end of a 1 week period, the percentage of Cs+ desorption from clinoptilolite did not exceed 7%, while it amounted to more than 30% in kaolinite, indicating more stable fixation by clinoptilolite. The sorption data were best described using Freundlich and D-R isotherm models. Sorption showed spontaneous and exothermic behavior on both minerals, with deltaH(0) being -6.3 and -11.4 kJ/mol for Cs+ uptake by kaolinite and clinoptilolite, respectively. Expanding the kaolinite interlayer space from 0.71 to 1.12 nm using DMSO intercalation, did not yield a significant enhancement in the sorption capacity of kaolinite, indicating that the surface and edge sites of the clay are more energetically favored. EDS mapping and elemental analysis of the surface of kaolinite and clinoptilolite revealed more intense signals on the surface of the latter with an even distribution of sorbed Cs+ onto the surfaces of both minerals. PMID- 15797387 TI - The F box protein Dsg1/Mdm30 is a transcriptional coactivator that stimulates Gal4 turnover and cotranscriptional mRNA processing. AB - We report here that the prototypical yeast transcription factor Gal4 undergoes two distinct modes of ubiquitin-mediated proteolysis: one that occurs independent of transcription and restricts Gal4 function, and another that is transcription coupled and essential for productive activation of Gal4 target genes. Destruction of transcriptionally active Gal4 depends on an F box protein called Dsg1/Mdm30. In the absence of Dsg1, Gal4 is stable, nonubiquitylated, and unable to productively stimulate transcription. Analysis of the phenotype of dsg1-null yeast reveals a striking disconnect between GAL gene RNA and protein levels; in the absence of Dsg1, Gal4 target genes are transcribed, but the resulting RNAs are not translated. The translational defects of these RNAs are related to defects in phosphorylation of the RNA polymerase II carboxy-terminal domain, which in turn affects recruitment of RNA processing machinery. We propose that Gal4 ubiquitylation and destruction are required for initiation-competent transcription complexes to transition to fully mature elongating complexes capable of appropriate mRNA processing. PMID- 15797390 TI - Supramolecular structures on silica surfaces and their adsorptive properties. AB - The study of adsorptive and chemical immobilization of beta-cyclodextrin on a surface of hydroxylated silicas with various porous structure is described. Using IR spectroscopy, thermal gravimetrical analysis with a programmed heating, and chemical analysis of the silica surface, it is shown that the process of adsorption-desorption of beta-cyclodextrin depends on the porous structure of the silica. The reaction of esterification was used for chemical grafting of beta cyclodextrin on the surface of hydroxylated silicas. Hydrolytic stability of silicas chemically modified by beta-cyclodextrin apparently is explained by simultaneous formation of chemical and hydrogen bonds between surface silanol groups and hydroxyl groups of beta-cyclodextrin. The uptake of the cations Cu(II), Cd(II), and Pb(II) and the anions Cr(VI) and As(V) by silicas modified with beta-cyclodextrin is investigated as a function of equilibrium ion concentrations. The increase of ion uptake and selectivity of ion extraction in comparison with starting silicas is established. It is due to the formation of surface inclusion complexes of the "host-guest" type in which one molecule of beta-cyclodextrin interacts simultaneously with several ions. PMID- 15797392 TI - Adsorption of cationic polyelectrolyte at the solid/liquid interface and dispersion of nanosized silica in water. AB - Adsorption of cationic polyelectrolyte, a homopolymer of maleimide propyl trimethylammonium chloride (MPTMAC), on silica nanoparticles from aqueous solution was studied. The adsorbed amount of MPTMAC and the adsorption layer thickness from solutions of different pH, polyelectrolyte concentration, salt type, and salt concentration were measured. The adsorbed amount exhibited a maximum as a function of the electrolyte concentration. The onset of the decline in the adsorbed amount depended on the type of counterions. The thickness of the adsorption layer increased gradually with increased of electrolyte concentration and leveled off at high electrolyte concentration. The enhanced adsorption in the presence of Na2SO4 can be explained by the bivalent SO4(2-) causing a better shielding effect. With increasing pH the adsorbed amount of MPTMAC increased, whereas the thickness of an adsorbed layer of MPTMAC decreased. At low polyelectrolyte concentrations unstable silica suspensions were observed from a stability test. At high polyelectrolyte concentrations the higher particle coverage caused electrosteric stabilization of the dispersion. However, further increase in MPTMAC concentration after saturated adsorption would flocculate the dispersed system. At low pH, MPTMAC tending to create a loops or tails conformation stabilized the suspension. PMID- 15797391 TI - Adsorption behavior of herbicide butachlor on typical soils in China and humic acids from the soil samples. AB - Three kinds of soils in China, krasnozem, fluvo-aquic soil, and phaeozem, as well as the humic acids (HAs) isolated from them, were used to adsorb the herbicide butachlor from water. Under the experimental conditions, the adsorption amount of butachlor on soils was positively correlated with the content of soil organic matter. HAs extracted from different kinds of soils had different adsorption capacity for the tested herbicide, which was positively correlated with their content of carbonyls. The adsorption mechanism was studied using Fourier transform infrared spectroscopy and cross-polarization with magic angle spinning 13C nuclear magnetic resonance (CP-MAS 13C NMR) techniques. It was showed that the adsorption mainly took place on the C=O, phenolic and alcoholic O-H groups of HAs. It was also confirmed that the adsorption mechanism was hydrogen bonds formation between the above groups of HAs and butachlor molecules. PMID- 15797393 TI - Particle and surface characterization of a natural illite and study of its copper retention. AB - Illite clays are known to have a strong affinity for metallic pollutants in the environment and can be applied as low-cost adsorbents for industrial waste treatment. A crucial factor in the development of such applications, however, is the understanding of the chemical, mineralogical, and colloidal properties of these clays. It is also important to understand the mechanisms involved in the surface adsorption of metals by these adsorbants. In order to study the retention of transition metals on illite clays, we have applied surface characterization techniques such as FPIA, SEM-EDX, XRD, N2 (77 K) adsorption, and FTIR. In addition to these experimental techniques, we have also employed a theoretical model that accounts for the chemistry of transition metal ions, and considers the global retention process to be the sum of several single retention processes. This model adequately fits the experimental data and allows for the speciation of metal retention on illite surfaces. Between pH values of 2.53 and 3.01 the only adsorption processes are the electrostatic sorption of [Cu(H2O)6]2+, and the surface complexation of [Cu(H2O)6]2+ and [Cu(OH)(H2O)5]+ ions. Surface complexation of [Cu(OH)(H2O)5]+ ions increases with pH, overcoming [Cu(H2O)6]2+ retention, and thus contributing to the surface precipitation of Cu(OH)2. PMID- 15797394 TI - Natural vermiculite as an exchanger support for heavy cations in aqueous solution. AB - The natural highly charged lamellar silicate vermiculite was investigated as an exchanger matrix in doubly distilled water solution to exchange magnesium inside the lamella with the heavy cations copper, nickel, cobalt, and lead at the solid/liquid interface. The extension of each exchange reaction was dependent on time of reaction, pH, and cation concentration. The maximum time presented the following order Pb2+ < Ni2+ < Cu2+ < Co2+, which corresponds to 12, 24, 48, and 72 h, respectively. The best performance was observed for nickel, as represented by the exchange capacity Nf, which gave values 0.59, 0.76, 0.84, and 0.93 mmol g( 1) for Pb2+ < Co2+ < Cu2+ < Ni2+, respectively. This capacity is dependent on pH interval variation from 1 to 9, being significantly increased in alkaline condition. The isotherm data were adjusted to a modified Langmuir equation and from the data the spontaneous Gibbs free energy was calculated. Linear correlations were obtained through Gibbs free energy or the maximum capacity against the cationic radius plot, with the lowest values for the largest cation lead. An exponential correlation was also observed for the maximum capacity versus enthalpy of hydration plot, indicating a difficulty of the less hydrated cation, lead, in exchanging with magnesium inside the lamellar space, as suggested by the proposed mechanism. The saturated matrices with cations presented a decrease in interlayer distance in comparison with the original vermiculite, which can be related to the hydrated phases, characteristic for each cation, with a lowest value for lead. PMID- 15797395 TI - Water vapor adsorption on activated carbon preadsorbed with naphtalene. AB - The adsorption of water vapor on a microporous activated carbon derived from the carbonization of coconut shell has been studied. Preadsorption of naphthalene was used as a tool to determine the location and the influence of the primary adsorbing centers within the porous structure of active carbon. The adsorption was studied in the pressure range p/p0=0-0.95 in a static water vapor system, allowing the investigation of both kinetic and equilibrium experimental data. Modeling of the isotherms using the modified equation of Do and Do was applied to determine the effect of preadsorption on the mechanism of adsorption. PMID- 15797397 TI - Preparation and characterization of electrically conducting Langmuir-Blodgett films of poly(N-octadecylaniline). AB - A stable monolayer of N-octadecylaniline containing stearic acid at the air-water interface has been obtained. However, the Langmuir monolayer of pure poly(N octadecylaniline) was not stable, but mixed Langmuir-Blodgett films of this polymer with stearic acid in different ratios as a spreading aid were stable. The electrical conductivity of these films increased by five orders of magnitude after doping with iodine as compared to that before iodine doping (5.8x10(-7) S cm(-1)). Temperature-dependent electrical conductivity suggested that these films were semiconducting in nature with low thermal activation energy. Impedance analyses of these films revealed that the equivalent circuit for poly(N octadecylaniline) was (RQ) whereas that for mixed poly(N-octadecylaniline) with stearic acid was (RQ)(RQ). PMID- 15797396 TI - Wetting fibers with liposomes. AB - Giant unilamellar vesicles (GUVs) are deposited on glass microfibers. The vesicles adopt the classical "onduloidal" shape of liquid droplets on fibers. They spread by two simultaneous mechanisms: envelopment and emission of a precursor film. This film spreads faster than on a uniform plane surface and eventually stops, signaling the presence of defects on the rod. This fast spreading tenses the vesicles; transient pores open on the GUVs and the internal liquid leaks out. This process leads to a new technique for fiber coating. PMID- 15797398 TI - Poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)-g poly(vinylpyrrolidone): association behavior in aqueous solution and interaction with anionic surfactants. AB - In this work, we aimed to study the association and interaction behavior of poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) block copolymers grafted with poly(vinylpyrrolidone). Critical micellization concentrations were determined using fluorescent probes (pyrene) and critical micellization temperatures characterizing temperature-dependent transitions from monomers to multimolecular micelles were measured. The thermal responsiveness of the copolymer is not affected by the grafting. The hydrodynamic radius of the graft copolymer micelles is found to be greater than that of the original copolymer micelles. The graft copolymer is found to form anisotropic aggregates. The structure of the graft copolymer micelles is less disrupted by the anionic surfactant sodium dodecyl sulfate, compared to the ungraft copolymer. PMID- 15797399 TI - Poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide)-g-poly(vinyl pyrrolidone): synthesis and characterization. AB - Pluronic poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) (PEO PPO-PEO) block copolymers are grafted with poly(vinyl pyrrolidone) by free radical polymerization of vinyl pyrrolidone with simultaneous chain transfer to the Pluronic in dioxane. This modified polymer has both thermal responsiveness and remarkable capacity to interact with a wide variety of hydrophilic and hydrophobic pharmaceutical agents which is very attractive for medical applications. The chemical structure of the graft copolymers was characterized by FTIR and 1H NMR spectroscopy. Polymerization conditions such as initiators, feed ratio, and reaction times are studied to obtain the ideal graft copolymer. PMID- 15797400 TI - Homogeneous forced hydrolysis of aluminum through the thermal decomposition of urea. AB - Homogeneous hydrolysis of aluminum by decomposition of urea in solution was achieved because the urea coordinates to the Al3+ in solution, forming [Al(H2O)5 (urea)]3+ and to a lesser extent [Al(H2O)4 (urea)2]3+. Upon hydrolysis more hydrolyzed monomeric species, [Al(H2O)5 (OH)]2+, [Al(H2O)4 (OH)2]+, [Al(H2O)4 (urea)(OH)]2+, and [Al(H2O)3 (urea)(OH)2]+, were formed, followed by trimeric species and the Al13 Keggin complex [AlO4Al12(OH)24(H2O)12]7+. The 27Al NMR spectra indicated the formation of other complexes in addition to the Al13 at the end of the hydrolysis reaction. PMID- 15797401 TI - Model cellulose films exposed to H. insolens glucoside hydrolase family 45 endo cellulase--the effect of the carbohydrate-binding module. AB - The effects of enzyme structure and activity on the degradation of model cellulose substrates were investigated by ellipsometry for the cellulase Humicola insolens GH45. The inactive variant D10N was found to adsorb at the cellulose surface but also to be incorporated into the cellulose films to an extent that depended on pH. For the native protein, the initial adsorption monitored for the inactive variant D10N was followed by enzyme-mediated degradation of the cellulose films. Again, a dependence on pH was found, such that higher pH resulted in slower enzymatic degradation. Removing the carbohydrate-binding module eliminated this pH dependence but also resulted in a decreased adsorption to the cellulose surface, and in a decreased net catalytic effect. PMID- 15797402 TI - Effect of thermal treatment on interfacial properties of beta-lactoglobulin. AB - The changes in the secondary conformation and surface hydrophobicity of beta lactoglobulin subjected to different thermal treatments were characterized at pH values of 7, 5.5 and 4 using circular dichroism (CD) and hydrophobic dye binding. Heating resulted in a decrease in alpha-helix content with a corresponding increase in random coil at all pH values, this change being more pronounced for small heating times. Heating also resulted in an increase in surface hydrophobicity as a result of partial denaturation, this increase being more pronounced at pH 4. Thermal treatment resulted in a shift of the spread monolayer isotherm at air-water interface to smaller area per molecule due to increased flexibility and more loop formation. Thermal treatment led to an increase in interfacial shear elasticity and viscosity of adsorbed beta-lactoglobulin layer at pH 5.5 and 7. Interfacial shear elasticity, shear viscosity, stability of beta lactoglobulin stabilized emulsion and average coalescence time of a single droplet at a planar oil-water interface with adsorbed protein layer exhibited a maximum for protein subjected to 15 min heat treatment at pH 7. At pH 5.5, the interfacial shear rheological properties and average single drop coalescence time were maximum for 15 min heat treatment whereas emulsion stability was maximum for 5 min heat treatment. At pH 7, thermal treatment was found to enhance foam stability. Analysis of thin film drainage indicated that interfacial shear rheological properties do not influence thin film drainage. PMID- 15797403 TI - A mechanistic study of the permeation kinetics through biomembrane models: gemcitabine-phospholipid bilayer interaction. AB - The kinetics of the interaction between Gemcitabine (a new anticancer drug) and phospholipid membrane models was investigated. This kind of study is of particular importance both in hypothesizing the interaction of Gemcitabine with mammalian cell membranes and in evaluating the potentiality of liposomes as a Gemcitabine delivery system. Unilamellar (LUV) and multilamellar (MLV) membrane models were made up of dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphatidic acid sodium salt (DMPA), or a DMPC-DMPA mixture (1:1 molar ratio). Gemcitabine-phospholipid vesicle interaction was studied by differential scanning calorimetry (DSC) measurements performed at different time intervals. The findings showed slower permeation kinetics of Gemcitabine through MLV than LUV which, at the same lipid/water ratio, are characterized by a larger lipid surface in contact with the drug aqueous solution. Another interesting difference between LUV and MLV is the onset of a transient two-peak structure during the DSC scans of MLVs. The effect is due to the unequal distribution of the drug between the outer and inner bilayers of the multilamellar vesicles during the permeation kinetics. At equilibrium the two-peak structure merges into a unique peak. This finding may provide useful information about the lipid bilayer permeability in model membranes. PMID- 15797404 TI - Penetration and interactions of the antimicrobial peptide, microcin J25, into uncharged phospholipid monolayers. AB - Microcin J25 forms stable monolayers at the air-water interface showing a collapse at a surface pressure of 5 mN/m, 220 mV of surface potential, and 6 fV per squared centimeter of surface potential per unit of molecular surface density. The adsorption of microcin J25 from the subphase at clean interfaces leads to a rise of 10 mN/m in surface pressure and a surface potential of 220 mV. From these data microcin appears to be a poor surfactant per se. Nevertheless, the interaction with the lipid monolayer further increase the stability of the peptide at the interface depending on the mode in which the monolayer is formed. Spreading with egg PC leads to nonideal mixing up to 7 mN/m, with hyperpolarization and expansion of components at the interface, with a small excess free energy of mixing caused by favorable contributions to entropy due to molecular area expansion compensating for the unfavorable enthalpy changes arising from repulsive dipolar interactions. Above 7 mN/m microcin is squeezed out, leaving a film of pure phospholipid. Nevertheless, the presence of lipid at 10 and 20 mN/m stabilize further microcin at the interface and adsorption from the subphase proceeds up to 30 mN/m, equivalent to surface pressure in bilayers. PMID- 15797406 TI - Characterizations and properties of hairy latex particles. AB - Industrial latex composed of a hydrophobic core surrounded by a charged hydrophilic layer exhibits excellent stability toward monovalent salt. That feature is classically attributed to a steric effect due to a loss of entropy during overlapping of coating materials. The so-called electrosteric stabilization is, however, not a straightforward function of the nature of the hydrophilic corona. This suspension was characterized in dilute solution by scattering and electrophoresis techniques. In contrast to spherical brushes the interface between the core and the corona is not well defined. The layer is more similar to a highly hydrated nonuniform gel with few longer strands that control the hydrodynamic properties than to a polyelectrolyte brush whose dependence on ionic strength reflects the concentration of counterions inside a well-defined structure. Thus the steric contribution to stabilization of these hairy particles appears to be insignificant in the range studied. The highly hydrated nature and the global charge of the layer are two predominant factors for the stability of the particles. PMID- 15797405 TI - Atomic force microscopy measurement of the elastic properties of the kidney epithelial cells. AB - Direct interaction force measurements using atomic force microscopy (AFM) were carried out between a silicon nitride tip and renal epithelial cells (Madin-Darby Canine Kidney-MDCK and proximal tubular epithelial cells derived from pig kidneys, LLC-PK1). The approaching (extending) portion of the force/distance curves is considered, and repulsive forces in the long range of 2-3 microm were seen in both MDCK as well as LLC-PK1 cells growing under normal conditions. The repulsive force in the shorter distance range of 50-200 nm was also observed, when cells were damaged exposing the underlying basal membrane. LLC-PK1 cells were more prone to damage than the MDCK cells, hence short-range forces were common in the former and long-range forces in the latter cells. The functional dependence of repulsive force on the indentation depth changes, at small indentation depth the force increases linearly, while at larger indentations the force is a quadratic function of the distance, which is attributed to the elasticity of the membrane and the solid-like response of cells, respectively. The oxalate treatment of cells for 2-4 h gives rise to an increase in the elastic modulus of the cells. PMID- 15797407 TI - A simple and controlled method of preparing uniform Ag midnanoparticles on Tollens-soaked silica spheres. AB - Ag midnanoparticles (midnanoparticles are those particles whose diameters are in the range from 20 to 80 nm) with average size of 30-50 nm and tunable packing densities were formed on the surface of preformed Tollens-soaked silica spheres by a simple and controlled method. The process mainly involved two steps. In the first step the absorption of Ag(NH3)2(+) ions occurred on the silica spheres and in the second step Ag(NH3)2(+) ions on the silica spheres were reduced to Ag midnanoparticles in the presence of glucose solution. The amount of Ag midnanoparticles on the silica spheres could easily be tuned by varying the washing times in the process of preparing the Tollens-soaked silica spheres. The washing process also effectively avoided the reduction of Ag(NH3)2(+) ions and the nucleation of Ag particles in solution and easily produced more uniform Ag midnanoparticles on the silica spheres. Attributing to the uniform Ag midnanoparticles, the Ag midnanoparticle-coated silica spheres show unique optical properties in the UV-vis absorption spectra. The resulting Ag midnanoparticle-coated silica spheres were characterized with transmission electron microscopy, UV-vis-IR recording spectrophotometry, and X-ray photoelectron spectroscopy. PMID- 15797408 TI - Gel trapping of dense colloids. AB - Phase density differences in sols, foams, or emulsions often lead to sedimentation or creaming, causing problems for materials where spatial uniformity over extended periods of time is essential. The problem may be addressed through the use of rheology modifiers in the continuous phase. Weak polymer gels have found use for this purpose in the food industry where they appear to be capable of trapping dispersoid particles in a three-dimensional matrix while displaying water-like viscosities at low shear. Attempts to predict sedimentation stability in terms of particle properties (size, shape, density difference) and gel yield stress have led to qualitative success for suspensions of large particles. The effect of particle size, however, in particular the case in which colloidal dimensions are approached, has not been investigated. The present work seeks to determine useful stability criteria for colloidal dispersions in terms of readily accessible viscoelastic descriptors. Results are reported for systems consisting of 12 microm poly(methyl methacrylate) (PMMA) spheres dispersed in aqueous gellan gum. Monovalent salt concentration is varied to control rheological properties, and sedimentation/centrifugation experiments are performed to determine dispersion stability. Necessary conditions for stability consist of a minimum yield stress together with a value of tan delta less than unity. PMID- 15797409 TI - Synthesis of dendrimer-protected TiO2 nanoparticles and photodegradation of organic molecules in an aqueous nanoparticle suspension. AB - Dendrimer-protected TiO2 nanoparticles were synthesized by hydrolysis of TiCl4 in solutions of poly(amido amine) dendrimers (64 terminals) under cooling. The morphology of dendrimers surrounding TiO2 nanoparticles depended on the terminal groups (amine, carboxyl, hydroxy) of dendrimers. The size (4.4-6.7 nm) of dendrimer-protected TiO2 nanoparticles was slightly smaller than that (7.5 nm) of bare TiO2 nanoparticles. The photodegradation of 2,4-dichlorophenoxyacetic acid revealed that dendrimer-protected TiO2 nanoparticles are more active as a photocatalyst than TiO2 nanoparticles without protectors. This suggests that the dendrimer acts as a reservoir of photoreacting reagents besides acting as a protector of nanoparticles. PMID- 15797410 TI - Nanofibers of LiMn2O4 by electrospinning. AB - Through sol-gel processing and electrospinning technique, extrathin fibers of poly(vinyl alcohol) (PVA)/lithium chloride/manganese acetate composite fibers were prepared. After calcination of the above precursor fibers at 600 degrees C, the spinel lithium manganese oxide (LiMn2O4) nanofibers, with a diameter of 100 200 nm, were successfully obtained. The fibers were investigated by TG-DTA, XRD, FT-IR, and SEM, respectively. The results showed that the crystalline phase and morphology of the fibers were largely influenced by the calcination temperature. PMID- 15797411 TI - CFD simulation of shear-induced aggregation and breakage in turbulent Taylor Couette flow. AB - An experimental and computational investigation of the effects of local fluid shear rate on the aggregation and breakage of approximately 10 microm latex spheres suspended in an aqueous solution undergoing turbulent Taylor-Couette flow was carried out. First, computational fluid dynamics (CFD) simulations were performed and the flow field predictions were validated with data from particle image velocimetry experiments. Subsequently, the quadrature method of moments (QMOM) was implemented into the CFD code to obtain predictions for mean particle size that account for the effects of local shear rate on the aggregation and breakage. These predictions were then compared with experimental data for latex sphere aggregates (using an in situ optical imaging method). Excellent agreement between the CFD-QMOM and experimental results was observed for two Reynolds numbers in the turbulent-flow regime. PMID- 15797412 TI - Lateral separation of colloids or cells by dielectrophoresis augmented by AC electroosmosis. AB - Colloidal particles and biological cells are patterned and separated laterally adjacent to a micropatterned electrode array by applying AC electric fields that are principally oriented normally to the electrode array. This is demonstrated for yeast cells, red blood cells, and colloidal polystyrene particles of different sizes and zeta-potentials. The separation mechanism is observed experimentally to depend on the applied field frequency and voltage. At high frequencies, particles position themselves in a manner that is consistent with dielectrophoresis, while at low frequencies, the positioning is explained in terms of a strong coupling between gravity, the vertical component of the dielectrophoretic force, and the Stokes drag on particles induced by AC electroosmotic flow. Compared to high frequency dielectrophoretic separations, the low frequency separations are faster and require lower applied voltages. Furthermore, the AC electroosmosis coupling with dielectrophoresis may enable cell separations that are not feasible based on dielectrophoresis alone. PMID- 15797413 TI - Effect of electrolyte on surface free energy components of feldspar minerals using thin-layer wicking method. AB - Application of the thin-layer wicking (TLW) technique on powdered minerals is useful for characterizing their surfaces. Albite (Na-feldspar) and orthoclase (K feldspar) are feldspar minerals which are frequently found in the same matrix. Despite similarities in their physicochemical properties, separation of these minerals from each other by flotation is generally possible in the presence of monovalent salts such as NaCl. Both albite and orthoclase exhibit the same microflotation properties and rather close electrokinetic profiles in the absence of salt. In this study, contact angles of albite and orthoclase determined by the TLW technique yielded close values in the absence and presence of amine collector. While the calculated surface energies and their components determined using contact angle data reveal that the energy terms remain farther apart in the absence of the collector, the differences narrow down at collector concentrations where full flotation recoveries are obtained. However, the effect of addition of NaCl on contact angles and surface free energy components at constant amine concentration indicates that albite is significantly affected by salt addition, whereas orthoclase remains marginally affected. This interesting finding is explained on the basis of ion-exchange properties, the stability of the interface, flotation data, and zeta potential data in the presence of NaCl. PMID- 15797414 TI - Interface characteristics in diffusion bonding of Fe3Al with Cr18-Ni8 stainless steel. AB - Fe3Al and Cr18-Ni8 stainless steel were diffusion-bonded in vacuum and a Fe3Al/Cr18-Ni8 interface with reaction layer was formed. Microstructure in the reaction layer at Fe3Al/Cr18-Ni8 interface was analyzed by means of scanning electron microscope (SEM) and electron probe micro-analyzer (EPMA). The growth of reaction layer with heating temperature (T) and holding time (t) was researched. The results indicate that FeAl, Fe3Al, Ni3Al, and alpha-Fe (Al) solid solution are formed in the reaction layer. These phases are favorable to promote the element diffusion and to accelerate the formation of the reaction layer at Fe3Al/Cr18-Ni8 interface. The growth of reaction layer obeys the parabolic law and its thickness (X) is expressed by X2 = 7.5 x 10(-4)exp(-83.59/RT)(t - t0). PMID- 15797415 TI - Characterisation of the protolytic properties of synthetic carbonate free fluorapatite. AB - The acid/base surface properties of carbonate free fluorapatite (Ca5(PO4)3F) have been characterised using high precision potentiometric titrations and surface complex modelling. Synthetic carbonate free fluorapatite was prepared and characterised by SEM, XRD, FT-IR and FT-Raman. The specific surface area was determined to be 17.7+/-1.2 m2 g(-1) with BET (N2 adsorption). The titrations were performed at 25+/-0.2 degrees C, within the pH range 5.7-10.8, in 0.10 and 0.50 mol dm(-3) NaNO3 ionic media. Experimental data were interpreted using the constant capacitance model and the software FITEQL 4.0. The surface equilibria: [triple bond]S1OH <==> [triple bond]S1O- + H+ lg betaS(-110) (int), [triple bond]S2OH <==> [triple bond]S2O- + H+ lg betaS(-101) (int) well describes the surface characteristics of synthetic fluorapatite. The equilibrium constants obtained were: lg betaS(-110) (int) = -6.33+/-0.05 and lg betaS(-101) (int) = 8.82+/-0.06 at I = 0.10 mol dm(-3). At the ionic strength 0.50 mol dm(-3), the equilibrium constants were slightly shifted to: lg betaS(-110) (int) = -6.43+/ 0.05 and lg betaS(-101) (int) = -8.93+/-0.06. The number of active surface sites, N(s), was calculated from titration data and was found to be 2.95 and 2.34 sites nm(-2) for the ionic strengths 0.10 and 0.50 mol dm(-3), respectively. pH(PZC) or the IEP was found to be 5.7 from Z-potential measurements. PMID- 15797416 TI - Surface complex characteristics of synthetic maghemite and hematite in aqueous suspensions. AB - The acid-base properties of the maghemite (gamma-Fe2O3)/water and hematite (alpha Fe2O3)/water interfaces have been studied by means of high precision potentiometric titrations and the experimental results are evaluated as surface complexation reactions. Synthetic maghemite and hematite were prepared and characterized using a combination of SEM, FT-IR and XRD. The specific surface area of the minerals was determined by the BET method. The titrations were performed at 25.0+/-0.2 degrees C within the range 2.8 0.05). Functional and subjective measures also revealed no pre and post treatment differences between the groups (P > 0.05), except for improved knee range of motion in the placebo group (P < 0.05). CONCLUSION: Joint inflammation in knee osteoarthritis, measured using radioleucoscintigraphy, was not altered significantly by pulsed shortwave, therefore this therapeutic modality has little or no anti-inflammatory effect on conditions such as osteoarthritis of the knee. PMID- 15797495 TI - Study of serum factors potentially involved in the pathogenesis of heterotopic bone formation after severe brain injury. AB - OBJECTIVE: To look for serum factors detectable early after head injury and predictive of heterotopic bone formation. PATIENTS AND METHODS: In this prospective study of a homogeneous population of 31 men with severe brain injury, blood samples were obtained 3 months after the accident, and levels of serum factors influenced by bone metabolism were compared between patients with and without heterotopic bone formation. As extensive fractures can influence serum factors, the patients without heterotopic bone formation were divided into two groups based on whether they had major fractures. Radionuclide bone scanning was used to validate patient classification. RESULTS: The group with heterotopic bone formation had significantly higher serum alkaline phosphatase levels (P < 0.01) and significantly lower serum leptin levels (P < 0.01), as compared to the other two groups. Body mass index and serum creatinine were comparable in the three groups. CONCLUSION: Leptin may be associated with the development of heterotopic bone formation. The antiosteogenic effect of leptin mediated by hypothalamic neurons may be impaired by hypothalamic damage related to severe brain injury. PMID- 15797497 TI - Suspected disk space infection with negative microbiological tests-report of eight cases and comparison with documented pyogenic discitis. AB - Few data are available on patients with suspected noniatrogenic pyogenic discitis but negative microbiological tests. OBJECTIVES: To compare the features, treatment, and outcomes in patients with suspected versus microbiologically documented noniatrogenic pyogenic discitis. PATIENTS: A retrospective chart review identified eight patients with suspected noniatrogenic pyogenic discitis managed at our institution over a 15-year period. Eighteen age- and sex-matched patients with microbiologically documented noniatrogenic pyogenic discitis managed at our institution during the same period served as controls. RESULTS: The eight cases had a longer time to diagnosis, a greater risk of abscess formation or epidural infection, and a smaller number of cutaneous portals of entry, as compared to the controls. None of the cases had evidence of endocarditis or diabetes mellitus. Infection at another site preceded the discitis in half the cases. Empirical treatment with two antimicrobials (usually a fluoroquinolone with a beta-lactam or fosfomycin) ensured a full recovery in all eight cases, with no relapses or long-term recurrences, whereas relapses occurred in three of the 18 controls treated with antibiotics selected by antibiotic susceptibility testing. CONCLUSION: Noniatrogenic pyogenic discitis with negative microbiological studies is associated with distinctive clinical features consistent with smoldering infection. The outcome is favorable under empirical two-drug antimicrobial therapy, usually including a fluoroquinolone given by the intravenous route. PMID- 15797498 TI - Role and modalities of information and education in the management of patients with rheumatoid arthritis: development of recommendations for clinical practice based on published evidence and expert opinion. AB - OBJECTIVES: To develop recommendations for the information and education of patients with rheumatoid arthritis (RA) seen in everyday practice, using evidence from the literature, supplemented with expert opinion when needed. METHODS: A scientific committee developed eight questions using the Delphi consensus procedure. A task force reviewed the literature for answers to these questions, using the PubMed Medline database (1980-2004) and the 2002-2004 databases of the annual meetings held by the French Society for Rheumatology (SFR), the European League Against Rheumatism (EULAR), and the American College of Rheumatology (ACR); the indexing terms for the search were rheumatoid, arthritis, patient, education, information, knowledge, general practitioner, family doctor, and continuing medical education. Only articles in French or English were included. A panel of rheumatologists used the evidence thus compiled to develop recommendations for each question; gaps in evidence were filled by calling on the panelists' expert opinion. For each recommendation, the level of evidence and extent of agreement among panelists were specified. RESULTS: There were four general questions about the objectives, supports, and mode of delivery (group or one-on-one) of patient information and education, as well as on evaluating knowledge, and four specific questions on program content. The search identified 1235 articles; 144 were selected on the title and 118 of those on the abstract. Three abstracts presented at meetings were also kept. The evidence from the literature was presented to the panelists during interactive workshops. The panelists then developed eight recommendations, all of which were grade D because no published studies specifically addressed everyday clinical practice. Agreement among panelists ranged across recommendations from 85.7% to 100%. CONCLUSION: Recommendations about educating and informing patients with RA in everyday practice were developed. They should increase practice uniformity and ultimately optimize the management of patients with RA. PMID- 15797499 TI - Salmonella typhi osteitis can mimic tuberculosis. A report of three cases. AB - We report three cases of Salmonella typhi osteitis. One patient was an immunocompetent woman with a single focus of osteitis, another had heterozygous sickle cell disease with multifocal osteitis, and the remaining patient had acute discitis. Tuberculosis was considered in all three patients, based on origin from an endemic area (sub-Saharan Africa), a chronic course in the first two patients, and granulomas in a biopsy specimen from one patient. PMID- 15797500 TI - Protracted fever of unknown origin as the presenting symptom of Behcet's disease. Report of a case. AB - We report a case of Behcet's disease that presented as protracted fever of unknown origin. The diagnosis was established when a thromboembolic event and ora3l aphthous ulcers occurred simultaneously. Antibiotics had no effect on the fever, which resolved when glucocorticoid and anticoagulant therapy was given. Among causes of protracted fever of unknown origin, Behcet's disease is exceedingly rare but should be considered together with the other vasculitides. Above all, the presence of a fever should prompt a search for a thromboembolic complication. PMID- 15797501 TI - Spinal cord compression due to vertebral osteochondroma: report of two cases. AB - Osteochondroma, or exostosis, is the most common of all benign bone tumors. Spinal osteochondromas are uncommon but may cause neurological compromise. We report two cases of spinal cord compression by osteochondromas. One patient was a 17-year-old man with hereditary multiple exostoses who was presented with spastic paraparesis, a sensory level at T3-T4, and a pyramidal syndrome. Vertebral exostosis was suspected by magnetic resonance imaging and confirmed by histological examination. Surgical decompression was followed by complete resolution of the neurological impairments. The other patient was a 19-year-old man with spastic paralysis of the right lower limb and a pyramidal syndrome. Whereas magnetic resonance imaging suggested a neurofibroma, histological features were those of osteochondroma. Nine months elapsed from symptom onset to surgery. This delay led to residual neurological impairments, which resolved almost completely after rehabilitation therapy. Vertebral osteochondromas contribute only 1.3-4.1% of all osteochondromas. The lesion may be solitary or a manifestation of hereditary multiple exostosis. Magnetic resonance imaging shows the exact location of the lesion, most notably with relation to neighboring neurological structures. Spinal cord compression is uncommon and usually has a favorable outcome provided surgical decompression is performed before major neurological damage develops. PMID- 15797502 TI - Post-lumbar puncture arachnoiditis. The need for directed questioning. AB - The inflammation of the arachnoid mater may produce a fibrinous exudate around the roots that causes them to adhere to the dural sheath. We report the case of a man aged 23 years who suffered from acute inflammatory truncated sciatica. The diagnosis of adhesive arachnoiditis was made in front of clinical arguments associated to typical signs on Myelo CT Scan and MRI. The only explanation ever found was a traumatic lumbar puncture at the age of 6 years for suspected meningitis. Sequelae of arachnoiditis are difficult to diagnosis. When MRI or myelography suggests it as a possibility, precise directed questioning is necessary to seek a history, albeit distant, of spinal or meningeal events. PMID- 15797503 TI - Dysplasia epiphysealis hemimelica in a young girl: role of MRI in the diagnosis and follow-up. AB - This report describes a sporadic case of dysplasia epiphysealis hemimelica that developed in the proximal tibia of a 21-month-old girl. Three years after the surgical intervention the patient has made complete clinical recovery with a normal range of motion, a walk with no limping or pain, no leg length discrepancy or angular knee deformity. Even though the proximal tibia does not represent an infrequently involved site, we report the clinical, pathological and radiological features of our case both for the extreme rarity of dysplasia epiphysealis hemimelica and the very young age of the patient. The authors underline also the role of magnetic resonance imaging in the diagnosis, management and follow-up of this very rare condition. PMID- 15797504 TI - Low back pain and sciatica as the presenting symptoms of neurinoma near the conus medullaris. Contribution of magnetic resonance imaging. PMID- 15797505 TI - Chronic polyarthritis as the presenting manifestation of acute leukemia. PMID- 15797506 TI - Keeping DCs awake by putting SOCS1 to sleep. AB - It has recently been shown that small interfering RNA (siRNA)-mediated inhibition of suppressors of cytokine signalling 1 (SOCS1) function enhances the immunostimulatory capacity of dendritic cells (DCs), which leads to more effective cancer vaccination. This recent study demonstrates the importance of negative regulation of DC maturation and functions for DC-based immunotherapy. PMID- 15797507 TI - CD86 and beta2-adrenergic receptor stimulation regulate B-cell activity cooperatively. AB - IgG1 functions to neutralize and clear foreign antigens, such as extracellular bacteria. Therefore, it is important to understand the multiple mechanisms by which the level of IgG1) is regulated to maintain immune system and host homeostasis. Recent data show that the level of IgG1 produced by a B cell is increased following CD86 and beta2-adrenergic receptor (beta2AR) stimulation, through increased expression of the transcription factor Oct-2 and its coactivator OCA-B (Oct coactivator from B cells), respectively. This finding suggests that signaling pathways that are activated by an immunoreceptor (CD86) and a neuroreceptor (beta2AR) converge to regulate the IgG1 response. PMID- 15797508 TI - Specific memory within innate immune systems. AB - Research into immune defense has been considerably enriched by the increasing focus on innate immunity. This type of immunity is still considered to lack specific memory, largely because there is no evidence of mechanisms that could provide such memory (such as acquired immunity). However, recent experimental data demonstrate specific memory phenomena in invertebrates: these organisms are thought to rely solely on innate defense. Here, I argue that a clear definition of the terms 'specificity' and 'memory', together with dissection of the evolutionary roots of immune defense, show us that innate immunity should not be, and is probably not, necessarily free of specific memory. PMID- 15797509 TI - Regulators of the Toll and Imd pathways in the Drosophila innate immune response. AB - The innate immune response is the first line of defense against microbial infections in both insects and mammals. Systematic analysis of the innate immune response in the model organism Drosophila melanogaster has provided important insights into the mechanisms of pathogen recognition and host response. Recognition of pathogen-associated molecules, such as peptidoglycans, stimulates the Toll and immune deficiency (Imd) pathways to induce antimicrobial responses. The Toll and Imd pathways are homologous to the mammalian Toll-like receptor (TLR) and tumor necrosis factor receptor (TNFR) signaling pathways, respectively, and are essential for Drosophila to survive infection. In this Review, we will discuss the recent genetic, genomic and RNA interference analyses that have unveiled additional intricacy in the Toll and Imd pathways. PMID- 15797510 TI - Immune subversion by Helicobacter pylori. AB - To maintain prolonged colonization of the human gastric mucosa, Helicobacter pylori must avoid both innate and adaptive immune responses. During its long coexistence with humans, it has evolved complex strategies to maintain a mild inflammation of the gastric epithelium while limiting the extent of immune effector activity. Severe disease, associated with bacterial colonization, might reflect loss of this control. Several mechanisms and the bacterial factors involved in immune subversion have, in recent years, been elucidated, thus opening the possibility of a better understanding of the pathogenicity of this microorganism. PMID- 15797511 TI - Src and Syk kinases: key regulators of phagocytic cell activation. AB - Src-family kinases and Syk tyrosine kinases have crucial roles in multiple leukocyte intracellular signaling pathways. In immunoreceptor-related pathways, these enzymes work together sequentially, with Src-family kinases phosphorylating specific protein substrates, which in turn recruit and activate Syk. Recent evidence indicates that several non-immunoreceptors also use Src-family kinases and Syk in this same fashion. In leukocyte integrin signaling, the interaction between the kinases is more complex, where they appear to act in a sequential manner but the mechanisms by which they are activated remain poorly defined. Elucidating the regulation of these tyrosine kinase-based signaling pathways in leukocytes remains an important goal in understanding how immune cells respond to the multitude of activating agents they encounter. PMID- 15797512 TI - An update on the role of translesion synthesis DNA polymerases in Ig hypermutation. AB - Several years have passed since the discovery of activation-induced cytosine deaminase (AID), the molecule responsible for triggering hypermutation of Ig genes. We now know that AID deaminates cytosines in the DNA encoding the variable portion of the Ig receptor, although an additional role in deaminating a regulatory mRNA transcript has not been ruled out. A major question that remains unanswered is how AID, a cytosine deaminase, causes mutations at both G:C and A:T base pairs. Mounting evidence suggests the involvement of a group of error-prone DNA polymerases known to bypass DNA lesions: the translesion synthesis (TLS) DNA polymerases. In this Review, we discuss the evidence for a role of TLS DNA polymerases in Ig hypermutation and argue that a major remaining challenge in our understanding of this mechanism is the recruitment of TLS DNA polymerases to the Ig locus following AID-mediated cytosine deamination. PMID- 15797513 TI - Cellular ligands of activating NK receptors. AB - Human natural killer (NK) cells are equipped with a series of surface receptors that recognise different cellular ligands on potential target cells. Some of these ligands [e.g. human leukocyte antigen (HLA) class I] prevent an NK-mediated attack by interacting with inhibitory NK receptors (e.g. killer Ig-like receptors). Other ligands interact with activating NK receptors that, once engaged, induce both cytotoxicity and lymphokine release. Tumour transformation (or viral infection) frequently results in downregulation of surface HLA class I molecules together with upregulation or de novo expression of ligands of triggering NK receptors. Thus, transformed cells can become highly susceptible to NK-mediated lysis. However, although NK cells use different means to identify and fight target cells, target cells have various strategies to hide themselves, and disarm or even confuse the immune system. PMID- 15797514 TI - MHC polymorphism: AIDS susceptibility in non-human primates. AB - One of the main host factors controlling resistance to disease appears to be the MHC. The recent poor results in HIV-1/AIDS Phase 3 vaccine field trials underline the importance of non-human primate models for AIDS. These models have been, and will continue to be, important for the definition of protective immune responses relevant to successful vaccine design because they supply essential information on the basic biology of lentivirus infections, mechanisms of resistance, escape and vaccine development. PMID- 15797515 TI - Application of immobilized enzyme reactor in on-line high performance liquid chromatography: a review. AB - This review summarizes all the research efforts in the last decade (1994-2003) that have been spent to the various application of immobilized enzyme reactor (IMER) in on-line high performance liquid chromatography (HPLC). All immobilization procedures including supports, kind of assembly into chromatographic system and methods are described. The effect of immobilization on enzymatic properties and stability of biocatalysts is considered. A brief survey of the main applications of IMER both as pre-column, post-column or column in the chemical, pharmaceutical, clinical and commodities fields is also reported. PMID- 15797516 TI - Gas chromatography-mass spectrometry identification of photoproducts of hexahydroquinoline derivatives: potential calcium channel antagonists. AB - Photodegradation products of hexahydroquinoline derivatives (HHQ) have been analysed with gas chromatography-mass spectrometry (GC-MS). The photodegradation was carried out under the conditions recommended in the first version of the document issued by the International Conference on Harmonization (ICH), currently in force in the studies of photochemical stability of drugs and therapeutic substances. The study was performed on the compounds having two chlorine atoms at different positions of the phenyl ring. Photodegradation of dichlorophenyl derivatives of HHQ resulted in formation of one or three photoproducts. The main product of their decomposition was aromatic compound formed as a result of dehydrogenation of the dihydropyridine ring. The most often observed fragmentation pathway of the photoproducts formed was elimination of methyl and methoxy radicals from the ester groups. The fragmentation of the photoproducts containing one chlorine atom at the ortho-position of the phenyl ring occurred through elimination of chlorine radical. PMID- 15797517 TI - Partition of whey milk proteins in aqueous two-phase systems of polyethylene glycol-phosphate as a starting point to isolate proteins expressed in transgenic milk. AB - Partitioning behaviour of the bovine whey proteins (bovine serum albumin, alpha lactoalbumin and beta lactoglobulin) and alpha-1 antitrypsin in aqueous two-phase systems prepared with polyethyleneglycol (molecular masses: 1000; 1500 and 3350) potassium phosphate was analysed. Bovine serum albumin and alpha lactoalbumin concentrated in the polyethyleneglycol rich phase with a partition coefficient of 10.0 and 27.0, respectively, while beta lactoglubulin and alpha-1 antitrypsin showed affinity for the phosphate-rich phase with a partition coefficient of 0.07 and 0.01, respectively. An increase of medium pH induced an increase of the partition coefficient of these proteins while the increase in polyethyleneglycol molecular mass induced the opposite behaviour. The system polyethyleneglycol 1500 pH 6.3 showed the best capacity for recovering the alpha-1 antitrypsin with a yield of 80% and a purification factor between 1.5 and 1.8 from an artificial mixture of the milk whey proteins and alpha-1 antitrypsin. The method appears to be suitable as a starting point to isolate proteins expressed in transgenic milk. PMID- 15797519 TI - Rapid assay for catechol-O-methyltransferase activity by high-performance liquid chromatography-fluorescence detection. AB - A rapid assay for measuring the activities of catechol-O-methyltransferase (COMT) is described. The method is based on high-performance liquid chromatography (HPLC)-fluorescence detection, and includes on-line extraction of catecholamines with a precolumn, separation of norepinephrine (NE) and normetanephrine (NMN) on an ODS column, electrochemical oxidation, and post-column fluorogenic derivatization using ethylenediamine. The method took less than 25 min for one sample, which is half that of the previous method and the sensitivity was similar. The intra-day assay precisions were 0.52-1.6%, and the inter-day assay precisions were 3.6-5.8% for rat liver and cerebral cortex (n = 5). The method is suitable for the rapid measurement of COMT activities of many biological samples. PMID- 15797518 TI - Protein expression profiling of CLL B cells using replicate off-line strong cation exchange chromatography and LC-MS/MS. AB - In this study we use replicate 2D-LC-MS/MS analyses of crude membranes from B cells derived from a patient with chronic lymphocytic leukemia (CLL) to examine the protein expression profile of CLL B cells. Protein identifications made by replicate 2D-LC-MS/MS analysis of tryptic peptides from detergent solubilized B cell membrane proteins, as well as replicate LC-MS/MS analysis of single off-line strong cation exchange chromatography (SCX) fractions, were analyzed. We show that despite the variance in SCX, capillary LC, and the data-dependent selection of precursor ions, an overlap of 64% between proteins identified in replicate runs was achieved for this system. PMID- 15797521 TI - Peptide profiling in epithelial tumor plasma by the emerging proteomic techniques. AB - The plasma peptide component (PPC) from ten melanoma (Mel), breast cancer (BC) and healthy individuals was examined by a combination of RP-HPLC, surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) and tandem mass spectrometry. A three peak pattern (2023, 2039, 2053.5 m/z) was primarily observed in melanoma. Two peaks (2236.1 and of 2356.3 m/z) were found only in BC samples. Fibrinogen alpha and inter-alpha-trypsin inhibitor heavy chain H4 fragments were absent in both tumor samples. PMID- 15797520 TI - Development of an assay method for the detection and quantification of protease and non-nucleoside reverse transcriptase inhibitors in plasma and in peripherical blood mononuclear cells by liquid chromatography coupled with ultraviolet or tandem mass spectrometry detection. AB - We present a simple chromatographic method to detect and quantify protease inhibitors (PI), metabolites and non-nucleoside reverse transcriptase inhibitors (NNRTIs) in human plasma of HIV-1 infected patients and in peripheral blood mononuclear cells (PBMCs) using either liquid chromatography coupled with ultraviolet (LC-UV) or liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). A solid-liquid extraction was carried out on 500 microl of plasma as pre-treatment. Calibration curve ranges were from 50 (100) to 5000 ng/ml (indinavir). PBMC pellets from 7 ml of blood were lysed with methanol/tris with a calibration curve ranging from 0.25 to 250 ng/pellet. Simple modifications in the mobile phase composition (slight increase of ammonium acetate concentration and addition of methanol for LC-UV) easily linked the two analytical systems. PMID- 15797522 TI - Measurement of homocysteine thiolactone hydrolase activity using high-performance liquid chromatography with fluorescence detection and polymorphisms of paraoxonase in normal human serum. AB - We developed a non-radioactive and sensitive assay method for measurement of the HTL hydrolase (HTLase) activity in biological samples, using OPA as a fluorescent post-labeling agent, l-homocysteine thiolactone (L-HTL) as the substrate, and HPLC to achieve rapid and selective separation of the substrate and product. The method was applied to measure the activity of HTLase in human, rabbit, rat and mouse serum samples. In addition, the correlation between the serum HTLase activity and PON1 polymorphisms in Japanese subjects was also investigated. The serum HTLase activity in humans, as determined by measurement of the enzyme activity in 22 subjects, was found to be in the range of 0.89-2.06 nmol/min mg protein, with a mean activity of 1.44 nmol/min mg protein. PMID- 15797523 TI - Specific method for determination of gefitinib in human plasma, mouse plasma and tissues using high performance liquid chromatography coupled to tandem mass spectrometry. AB - A rapid, sensitive and specific method was developed and validated using liquid chromatography-tandem mass spectrometry (LC/MS/MS) for determination of gefitinib in human plasma and mouse plasma and tissue. Sample preparation involved a single protein precipitation step by the addition of 0.1 mL of plasma or a 200 mg/mL tissue homogenate diluted 1/10 in human plasma with 0.3 mL acetonitrile. Separation of the compounds of interest, including the internal standard (d8) gefitinib, was achieved on a Waters X-Terra C18 (50 mm x 2.1 mm i.d., 3.5 microm) analytical column using a mobile phase consisting of acetonitrile-water (70:30, v/v) containing 0.1% formic acid and isocratic flow at 0.15 mL/min for 3 min. The analytes were monitored by tandem mass spectrometry with electrospray positive ionization. Linear calibration curves were generated over the range of 1-1000 ng/mL for the human plasma samples and 5-1000 ng/mL for mouse plasma and tissue samples with values for the coefficient of determination of > 0.99. The values for both within- and between-day precision and accuracy were well within the generally accepted criteria for analytical methods (< 15%). This method was subsequently used to measure concentrations of gefitinib in mice following administration of a single dose of 150 mg/kg intraperitoneally and in cancer patients receiving an oral daily dose of 250 mg. PMID- 15797524 TI - A novel assay method for theanine synthetase activity by capillary electrophoresis. AB - The determination of theanine has been performed by micellar electrokinetic capillary chromatography (MECC) using 2,4-dinitrofluorobenzene (DNFB) as a derivative reagent. To achieve the separation, a fused-silica capillary column was used with a borax buffer at 0.03 mol/L pH 9.8 (containing Brij35 and isopropanol) at 17 degrees C with detection wave length at 360 nm. The factors affecting the efficiency of the sample separation were examined simultaneously. A 40-min reaction at 35 degrees C between l-glutamate and ethylamine (with Tris-HCl buffer, pH 7.5) was investigated using the theanine synthetase from budding tea seeds. A novel method for the analysis of theanine synthetase activity based on MECC was established. The method shows mean recovery ranged from 87.1 to 105.3% and linearity ranged from 0.2 to 5.0 mmol/L. PMID- 15797525 TI - Determination of urinary nucleosides by direct injection and coupled-column high performance liquid chromatography. AB - A coupled-column liquid chromatographic method for the direct analysis of 14 urinary nucleosides is described. Efficient on-line clean-up and concentration of 14 nucleosides from urine samples were obtained by using a boronic acid substituted silica column (40 mm x 4.0 mm I.D.) as the first column (Col-1) and a Hypersil ODS2 column (250 mm x 4.6 mm I.D.) as the second column (Col-2). The mobile phases applied consisted of 0.25 mol/L ammonium acetate (pH 8.5) on Col-1, and of 25 mmol/L potassium dihydrogen phosphate (pH 4.5) on Col-2, respectively. Determination of urinary nucleosides was performed on Col-2 column by using a linear gradient elution comprising 25 mmol/L potassium dihydrogen phosphate (pH 4.5) and methanol-water (60:40, v/v) with UV detection at 260 nm. Urinary nucleosides analysis can be carried out by this procedure in 50 min requiring only pH adjustment and the protein precipitation by centrifugation of urine samples. Calibration plots of 14 standard nucleosides showed excellent linearity (r > 0.995) and the limits of detection were at micromolar levels. Both of intra- and inter-day precisions of the method were better than 6.6% for direct determination of 14 nucleosides. The validated method was applied to quantify 14 nucleosides in 20 normal urines to establish reference ranges. PMID- 15797526 TI - High performance liquid chromatographic determination of N-butyryl glucosamine in rat plasma. AB - PURPOSE: A high performance liquid chromatography (HPLC) method for determination in plasma of N-butyryl glucosamine (GLBU), a highly water-soluble compound with no chromophore was developed. METHOD: To 100 muL of plasma containing GLBU was added fucose as internal standard. GLBU and fucose were derivatized using 1 phenyl-3-methyl-5-pyrazolone in the presence of sodium hydroxide at 70 degrees C for 30 min. The solution was neutralized with hydrochloric acid and the excess derivatizing reagent was extracted with chloroform. The aqueous layer was injected into an isocratic HPLC system consisting of an autoinjector, a single pump and a UV detector set at 245 nm. Two different 25 cm reversed phase columns were used, a 4 and a 10 microm C(18) columns. The mobile phase was a mixture of phosphate buffer (pH 7) and acetonitrile (80:20), which was run through a pump at a flow rate of 1.0 mL/min at ambient temperature. RESULTS: Derivatized fucose and GLBU appeared 24 and 28 min, and at 34 and 37 min using 4 and 10 microm columns, respectively. The assay was linear over the range of 0.2-200 microg/mL with a limit of quantification of 0.2 and 1 microg/mL for the 4 and 10 microm columns, respectively. The method was applied to the determination of GLBU in rat plasma after oral administration of 233 mg/kg of GLBU. CONCLUSION: The present assay is precise, and accurate with sufficient sensitivity for pharmacokinetic studies following therapeutically relevant doses. PMID- 15797527 TI - Simple method for the simultaneous isolation and determination of fumonisin B1 and its metabolite aminopentol-1 in swine liver by liquid chromatography- fluorescence detection. AB - An analytical method based on high-performance liquid chromatography (HPLC) combined with fluorescence detection (FL) has been developed for the simultaneous determination of fumonisin B1 (FB1) and its totally hydrolized metabolite aminopentol-1 (AP1) in pig liver. The sample preparation is based on a single solid phase extraction (SPE). o-Phthalaldehyde (OPA) was used for pre-column derivatization before the programmed reversed-phase analysis on phenylhexyl column. The developed method shows good repeatibility for inter- and intra-day precision as well as adequate linearity of calibration curves (r2 was 0.9855 for FB1 and 0.9831 for AP1). Average recoveries from the matrix were 93.6% for FB1 and 95.3% for AP1. The limit of quantification (LOQ) in swine liver was 75 microg/kg for FB1 and 42 microg/kg for AP1. PMID- 15797528 TI - Validation of a liquid chromatography-mass spectrometry method to assess the metabolism of dextromethorphan in rat everted gut sacs. AB - A rapid, sensitive and selective liquid chromatography-mass spectrometry (LC-MS) method was developed for the simultaneous assay of dextromethorphan and its metabolites in tissue culture medium and its intestinal metabolism studied with the rat everted gut sac model. The method was validated in the concentration range of 0.1-2.5 microM (27.1 ng/mL-0.677 microg/mL) for dextromethorphan and 0.005-0.5 microM for dextrorphan and 3-methoxymorphinan (1.28 ng/mL-0.128 microg/mL) and 3-hydroxymorphinan (1.22 ng/mL-0.122 microg/mL). The limits of quantification (LOQ) were 0.0025 microM (12.5 fmoles, 3.4 pg, 5 microL injected) for dextromethorphan; 0.0025 microM for dextrorphan, 3-methoxymorphinan (24.9 fmoles, 6.4 pg injected), and 3-hydroxymorphinan (25.1 fmoles, 6.1 pg injected) with 10 microL injected. The detection of dextrorphan and 3-methoxymorphinan showed that both the P450 isoforms CYP3A and 2D were active in the intestinal mucosa and metabolised dextromethorphan during its passage across the mucosa. PMID- 15797529 TI - The rat brain hippocampus proteome. AB - The hippocampus is crucial in memory storage and retrieval and plays an important role in stress response. In humans, the CA1 area of hippocampus is one of the first brain areas to display pathology in Alzheimer's disease. A comprehensive analysis of the hippocampus proteome has not been accomplished yet. We applied proteomics technologies to construct a two-dimensional database for rat brain hippocampus proteins. Hippocampus samples from eight months old animals were analyzed by two-dimensional electrophoresis and the proteins were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The database comprises 148 different gene products, which are in the majority enzymes, structural proteins and heat shock proteins. It also includes 39 neuron specific gene products. The database may be useful in animal model studies of neurological disorders. PMID- 15797530 TI - Isolation and quantification of dinucleoside polyphosphates by using monolithic reversed phase chromatography columns. AB - In former studies, dinucleoside polyphosphates were quantified using ion-pair reversed-phase perfusion chromatography columns, which allows a detection limit in the micromolar range. The aim of this study was both to describe a chromatographic assay with an increased efficiency of the dinucleoside separation, which enables the reduction of analytical run times, and to establish a chromatographic assay using conditions, which allow MALDI-mass spectrometric analysis of the resulting fractions. We compared the performance of conventional silica reversed phase chromatography columns, a perfusion chromatography column and a monolithic reversed-phase C18 chromatography column. The effects of different ion-pair reagents, flow-rates and gradients on the separation of synthetic diadenosine polyphosphates as well as of diadenosine polyphosphates isolated from human platelets were analysed. Sensitivity and resolution of the monolithic reversed-phase chromatography column were both higher than that of the perfusion chromatography and the conventional reversed phase chromatography columns. Using a monolithic reversed-phase C18 chromatography column, diadenosine polyphosphates were separable baseline not only in the presence of tetrabutylammonium hydrogensulfate (TBA) but also in the presence of triethylammonium acetate (TEAA) as ion-pair reagent. The later reagent is useful because, in contrast to TBA, it is compatible with MALDI mass-spectrometric methods. This makes TEAA particularly suitable for identification of unknown nucleoside polyphosphates. Furthermore, because of the lower backpressure of monolithic reversed-phase chromatography columns, we were able to significantly increase the flow rate, decreasing the amount of time for the analysis close to 50%, especially using TBA as ion-pair reagent. In summary, monolithic reversed phase C18 columns markedly increase the sensitivity and resolution of dinucleoside polyphosphate analysis in a time-efficient manner compared to reversed-phase perfusion chromatography columns or conventional reversed-phase columns. Therefore, further dinucleoside polyphosphate analytic assays should be based on monolithic silica C18 columns instead of perfusion chromatography or conventional silica reversed phase chromatography columns. In conclusion, the use of monolithic silica C18 columns will lead to isolation and quantification of up to now unknown dinucleoside polyphosphates. These chromatography columns may facilitate further research on the biological roles of dinucleoside polyphosphates. PMID- 15797531 TI - Capillary electrophoretic determination of methotrexate, leucovorin and folic acid in human urine. AB - A simple, rapid and sensitive procedure using capillary zone electrophoresis (CZE) to measure methotrexate, folinic acid and folic acid in human urine has been developed and validated. Optimum separation of methotrexate, folinic acid and folic acid was obtained on a 60 cm x 75 microm capillary using a 15 mM phosphate buffer solution (pH 12.0), temperature and voltage 20 degrees C and 25 kV, respectively and hydrodynamic injection. Under these conditions the analysis takes approximately 9.0 min. Good results were obtained for different aspects including stability of the solutions, linearity, accuracy and precision. Before CZE determination, the urine samples were purified and enriched by means of a solid phase extraction step with a preconditioned C(18) cartridge and eluting the compound with a mixture 1:1 of methanol:water. A linear response over the urine concentration range 1.0-6.0 mgL(-1) for MTX and 0.5-6.0 mgL(-1) for folinic acid and folic acid was observed. Detection limits for the three compound in urine were 0.35 mgL(-1). CZE was shown to be a good method with regard to simplicity, satisfactory precision, and sensitivity. PMID- 15797532 TI - Optimized determination of lycopene in canine plasma using reversed-phase high performance liquid chromatography. AB - An isocratic high-performance liquid chromatographic method with detection at 472 nm was developed, optimized and validated for the determination of lycopene in canine plasma. Ethyl-beta-apo-8'-carotenoate was used as internal standard. A Hypersil BDS RP-C18 column (150 mm x 4.6 mm), 5 microm particle size, was equilibrated with a mobile phase composed of acetonitrile and methanol (50:50, v/v). Its flow rate was 1.5 ml/min. The elution time for lycopene and ethyl-beta apo-8'-carotenoate was approximately 11 and 5 min, respectively. Calibration curves of lycopene were linear in the concentration range of 3-200 ng/ml in plasma. Limits of detection and quantification in plasma were 1 and 4 ng/ml, respectively. Recovery was greater than 97%. Intra- and inter-day relative standard deviation for lycopene in plasma was less than 1.8 and 3.1%, respectively. This method was applied to the determination of lycopene plasma levels after single dose administration to dogs. PMID- 15797533 TI - Analysis of environmental biomarkers in urine using an electrochemical detector. AB - Phenols are present in the environment and are prevalent in human populations, as environmental contaminants, dietary components, or their metabolites. Many are potential endocrine-altering agents. Currently available methods analyze single components or single families of chemicals as biomarkers of exposure. In order to assess multiple biologically relevant exposures to such substances, we evaluated the feasibility of determining several phenols simultaneously in urine, using an electrochemical detector (ECD) in combination with high performance liquid chromatography (LC). Based on reported analyses in the literature and the ECD response, we selected four xenobiotic residues, including three phytoestrogens (enterolactone, daidzein, and genistein) and bisphenolA [BPA]. These compounds had detection limits below 1 microg/L in urine using the cleanup procedure (glucuronidase hydrolysis and C18 column) and the urine volume (2 mL) we employed. As a pilot study to demonstrate the method's utility, we determined urinary enterolactone, daidzein, genistein and BPA in samples from nine children and 24 adults. PMID- 15797534 TI - Determination of WR-1065 in human blood by high-performance liquid chromatography following fluorescent derivatization by a maleimide reagent ThioGlo3. AB - In order to improve the sensitivity and stability of human blood samples containing WR-1065 (i.e., active metabolite of the cytoprotective agent amifostine), a high-performance liquid chromatographic method was developed and validated using fluorescent derivatization with ThioGlo3. Using a sample volume of only 100 microl, the method was specific, sensitive (limit of quantitation=10 nM in deproteinized blood or 20 nM in whole blood), accurate (error < or = 3.2%) and reproducible (CV < or = 8.7%). In addition, the stability of WR-1065 in deproteinized and derivatized blood samples was assured for at least four weeks at -20 degrees C. This method should be particularly valuable in translating the kinetic-dynamic relationship of WR-1065 in preclinical models to that in cancer patients. PMID- 15797535 TI - Qualitative assessment of IC50 values of inhibitors of the neuronal nicotinic acetylcholine receptor using a single chromatographic experiment and multivariate cluster analysis. AB - It has been widely demonstrated that affinity chromatography can be used to derive binding affinities, and that these affinities can be correlated to data obtained using standard techniques such as membrane binding, ultrafiltration and equilibrium dialysis. The purpose of this study is to evaluate the use of immobilized nicotinic acetylcholine receptor stationary phase in chromatographic experiments to assess the functional activity of series of noncompetitive inhibitors (NCIs) as reflected in their IC50 values. Chromatographically determined retention values and computer generated molecular descriptors were obtained for 29 compounds and the data were analyzed by cluster analysis. The approach qualitatively ranked the test compounds as efficient NCIs (low IC50 values) or poor NCIs (high IC50 values). The data obtained with the 29 compounds used in this study demonstrate that the experimental approach had been able to place 25 of these compounds in the correct IC(50) clusters. To our knowledge, this is the first relationship established between chromatographic retention and IC50 for membrane-bound receptors. These results suggest that the chromatographic approach may be useful in development of lead drug candidates including the determination of off-target binding. PMID- 15797536 TI - A sensitive liquid chromatography-tandem mass spectrometry method for the quantification of mometasone furoate in human plasma. AB - A robust, rapid, selective and sensitive liquid chromatography-negative atmospheric pressure chemical ionization (LC-(APCI(-))-MS-MS) method has been developed for the quantification of mometasone furoate (MF) in human plasma utilizing a solid-phase extraction clean-up step and 13C-fluticasone propionate as internal standard. The intra- and inter-day coefficients of variation were < or = 15% and the lower limit of quantification (LLOQ) was 15 pg/ml. This method is ideally suited for pharmacokinetic investigations of low MF levels following inhalation of MF. PMID- 15797537 TI - Determination of deracoxib in feline plasma samples using high performance liquid chromatography. AB - A new HPLC procedure for the determination of deracoxib, a selective cyclooxygenase-2 inhibitor, has been developed and validated. Following a liquid liquid extraction using isopropyl alcohol and chloroform, samples were separated by isocratic reversed-phase HPLC on an Atlantis C18 column and quantified using UV detection at 252 nm. The mobile phase was a mixture of 10 mM potassium phosphate (pH 4.5) and acetonitrile, with a flow-rate of 1.0 ml/min. The procedure produced a linear curve over the concentration range 10-1500 ng/ml. The development of the assay allowed the determination of pharmacokinetic parameters after oral administration of deracoxib in cats and would be suitable for other pharmacokinetic studies. PMID- 15797538 TI - Rapid and sensitive liquid chromatography-tandem mass spectrometry method for the quantitation of levodropropizine in human plasma. AB - A rapid and sensitive LC-MS-MS method for quantifying levodropropizine in human plasma after oral administration of a single-dose (60 mg/day) was developed and validated. The sample preparation used liquid-liquid extraction with a mixture of dichloromethane-diethyl ether (2:3, v/v) in a basic environment. The retention time of levodropropizne and zolmitriptan (used as internal standard) was 1.6 and 1.4 min, respectively. The assay was linear over the range 0.25-500 ng/mL with a LOQ of 0.25 ng/mL. The intra- and inter-day precision were < 8.1% and < 11.5%, respectively, and the accuracy was in the range 87.6-112%. The levodropropizine concentration profile in human plasma was determined. PMID- 15797539 TI - Determination of lauroyl-indapamide in rat whole blood by high-performance liquid chromatography. AB - A method based on a liquid-liquid extraction procedure followed by high performance liquid chromatography (HPLC) coupled with UV-visible detection is described and validated for the determination of lauroyl-indapamide in rat whole blood. The blood sample was extracted with diethyl ether after the addition of 10% trifluoroacetic acid (aq.). The chromatographic separation was performed on a Chromasil ODS column, using methanol-acetonitrile-tetrahydrofuran-0.2% trifluoroacetic acid (170:20:15:38, v/v/v/v) as the mobile phase. The UV detection wavelength was set at 240 nm. The extraction recovery of lauroyl indapamide was ranged from 76.5 to 82.6%, and the calibration curve had a good linearity in the range of 0.048-200 microg/ml (r = 0.9976). The method presents appropriate intra-day and inter-days repeatabilities, showing values below 7.4% in terms of the percentage of relative standard deviation (R.S.D.). The method proposed is simple, rapid and sensitive, being useful for pharmacokinetic studies in rats. PMID- 15797540 TI - Determination of levetiracetam in human plasma with minimal sample pretreatment. AB - We here present a method for the routine quantification of the novel antiepileptic drug levetiracetam in human serum by HPLC-UV. The procedure is very easy, quick, inexpensive and rugged. The sample preparation consists only in the precipitation of serum proteins by perchloric acid and extraction of unpolar components by cyclohexane. The aqueous phase containing the analyte levetiracetam is injected onto a porous graphitic carbon analytical HPLC-column and separated by gradient elution with diluted phosphoric acid/acetonitrile. Detection is carried out at a wavelength of 205 nm. The calibration function is linear in the range of 1-75 microg/ml. The detection limit is 0.1 microg/ml. Using four quality control sample concentrations, the inter-day relative standard deviations (R.S.D.) are lower than 3% and the accuracies are better than 6%. The respective inter-day values are: R.S.D. < 4% and accuracies better than 2%. Frequently co administered antiepileptic drugs do not interfere with the assay. The method has been successfully applied to patient samples. PMID- 15797541 TI - The simple life (of cortical progenitors). AB - Asymmetric cell division plays a major role in the generation of cell diversity during development. In this issue of Neuron, Sun and colleagues present evidence that the epidermal growth factor receptor is asymmetrically distributed in mitotic cerebral cortical precursors, and the resulting unequal inheritance generates offspring with different responsiveness to growth factor and unique cell fates. PMID- 15797542 TI - What's next? Sequential movement encoding in primary motor cortex. AB - Complex actions often can be decomposed into sequences of individual movements. Primate medial motor areas (SMA and pre-SMA) have been shown to be key players in the concert of such sequential actions. In this issue of Neuron, Lu and Ashe show for the first time that neurons in primate M1-the ultimate output stage of cortex have anticipatory activity related to specific movement sequences. These findings challenge the traditional view of M1 as being a simple module for generating movements. PMID- 15797543 TI - The importance of individual differences in grapheme-color synesthesia. AB - In this issue of Neuron, Hubbard et al. show individual differences in how grapheme-color synesthetes perform on cognitive tasks. Importantly, these behavioral differences were correlated with fMRI measures. Such individual differences have important ramifications for synesthesia research. If individual differences are ignored, then synesthesia research will be characterized by erroneous conclusions and failures to replicate. PMID- 15797544 TI - You're perfect, now change--redefining the role of developmental plasticity. AB - The receptive field properties of neurons in the developing brain can in many cases be remarkably similar to those of adult neurons. This raises the question of why these same neurons need the capacity for such impressive developmental plasticity, most clearly demonstrated by the rewiring that occurs in response to sensory deprivation. The roles of developmental neuronal plasticity in the assimilation of neurons into a larger network, including temporal and cross-modal integration, are discussed. PMID- 15797545 TI - Activity-dependent matching of excitatory and inhibitory inputs during refinement of visual receptive fields. AB - The receptive field (RF) of single visual neurons undergoes progressive refinement during development. It remains largely unknown how the excitatory and inhibitory inputs on single developing neurons are refined in a coordinated manner to allow the formation of functionally correct circuits. Using whole-cell voltage-clamp recording from Xenopus tectal neurons, we found that RFs determined by excitatory and inhibitory inputs in more mature tectal neurons are spatially matched, with each spot stimulus evoking balanced synaptic excitation and inhibition. This emerges during development through a gradual reduction in the RF size and a transition from disparate to matched topography of excitatory and inhibitory inputs to the tectal neurons. Altering normal spiking activity of tectal neurons by either blocking or elevating GABA(A) receptor activity significantly impeded the developmental reduction and topographic matching of RFs. Thus, appropriate inhibitory activity is essential for the coordinated refinement of excitatory and inhibitory connections. PMID- 15797546 TI - Activity regulates positive and negative neurotrophin-derived signals to determine axon competition. AB - Developmental axon competition plays a key role in sculpting neural circuitry. Here, we have asked how activity and neurotrophins could interact to select one axon over another. Using compartmented cultures of sympathetic neurons, we show that, in the presence of NGF, local depolarization confers a competitive growth advantage on the depolarized axon collaterals and at the same time disadvantages the growth of unstimulated axons from the same and competing neurons. Depolarization mediates the competitive advantage by activating a CaMKII-MEK pathway, which converges to enhance local NGF-mediated downstream growth signals. Patterned electrical stimulation also acts via this pathway to enhance NGF promoted axonal growth. In contrast, the competitive disadvantage is due to BDNF secreted from and acting on the unstimulated, competing axons through p75NTR. Thus, activity regulates both positive and negative neurotrophin-derived signaling cascades to confer a competitive growth advantage on one axon versus another, thereby providing a cellular mechanism for developmental axon selection. PMID- 15797547 TI - Mouse model of multiple system atrophy alpha-synuclein expression in oligodendrocytes causes glial and neuronal degeneration. AB - Transgenic (Tg) mice overexpressing human wild-type alpha-synuclein in oligodendrocytes under the control of the 2,' 3'-cyclic nucleotide 3' phosphodiesterase (CNP) promoter are shown here to recapitulate features of multiple system atrophy (MSA), including the accumulation of filamentous human alpha-synuclein aggregates in oligodendrocytes linked to their degeneration and autophagocytosis of myelin. Significantly, endogenous mouse alpha-synuclein also accumulated in normal and degenerating axons and axon terminals in association with oligodendroglia and neuron loss and slowly progressive motor impairments. Our studies demonstrate that overexpression of alpha-synuclein in oligodendrocytes of mice results in MSA-like degeneration in the CNS and that alpha-synuclein inclusions in oligodendrocytes participate in the degeneration of neurons in MSA. PMID- 15797548 TI - Regulation of gene expression by lithium and depletion of inositol in slices of adult rat cortex. AB - Lithium inhibits inositol monophosphatase at therapeutically effective concentrations, and it has been hypothesized that depletion of brain inositol levels is an important chemical alteration for lithium's therapeutic efficacy in bipolar disorder. We have employed adult rat cortical slices as a model to investigate the gene regulatory consequences of inositol depletion effected by lithium using cytidine diphosphoryl-diacylglycerol as a functionally relevant biochemical marker to define treatment conditions. Genes coding for the neuropeptide hormone pituitary adenylate cyclase activating polypeptide (PACAP) and the enzyme that processes PACAP's precursor to the mature form, peptidylglycine alpha-amidating monooxygenase, were upregulated by inositol depletion. Previous work has shown that PACAP can increase tyrosine hydroxylase (TH) activity and dopamine release, and we found that the gene for GTP cyclohydrolase, which effectively regulates TH through synthesis of tetrahydrobiopterin, was also upregulated by inositol depletion. We propose that modulation of brain PACAP signaling might represent a new opportunity in the treatment of bipolar disorder. PMID- 15797549 TI - Asymmetric distribution of EGFR receptor during mitosis generates diverse CNS progenitor cells. AB - It has been debated whether asymmetric distribution of cell surface receptors during mitosis could generate asymmetric cell divisions by yielding daughters with different environmental responsiveness and, thus, different fates. We have found that in mouse embryonic forebrain ventricular and subventricular zones, the EGFR can distribute asymmetrically during mitosis in vivo and in vitro. This occurs during divisions yielding two Nestin+ progenitor cells, via an actin dependent mechanism. The resulting sibling progenitor cells respond differently to EGFR ligand in terms of migration and proliferation. Moreover, they express different phenotypic markers: the EGFRhigh daughter usually has radial glial/astrocytic markers, while its EGFRlow sister lacks them, indicating fate divergence. Lineage trees of cultured cortical glioblasts reveal repeated EGFR asymmetric distribution, and asymmetric divisions underlie formation of oligodendrocytes and astrocytes in clones. These data suggest that asymmetric EGFR distribution contributes to forebrain development by creating progenitors with different proliferative, migratory, and differentiation responses to ligand. PMID- 15797550 TI - Serotonin-induced regulation of the actin network for learning-related synaptic growth requires Cdc42, N-WASP, and PAK in Aplysia sensory neurons. AB - Application of Clostridium difficile toxin B, an inhibitor of the Rho family of GTPases, at the Aplysia sensory to motor neuron synapse blocks long-term facilitation and the associated growth of new sensory neuron varicosities induced by repeated pulses of serotonin (5-HT). We have isolated cDNAs encoding Aplysia Rho, Rac, and Cdc42 and found that Rho and Rac had no effect but that overexpression in sensory neurons of a dominant-negative mutant of ApCdc42 or the CRIB domains of its downstream effectors PAK and N-WASP selectively reduces the long-term changes in synaptic strength and structure. FRET analysis indicates that 5-HT activates ApCdc42 in a subset of varicosities contacting the postsynaptic motor neuron and that this activation is dependent on the PI3K and PLC signaling pathways. The 5-HT-induced activation of ApCdc42 initiates reorganization of the presynaptic actin network leading to the outgrowth of filopodia, some of which are morphological precursors for the learning-related formation of new sensory neuron varicosities. PMID- 15797551 TI - Calcium-permeable AMPA receptor plasticity is mediated by subunit-specific interactions with PICK1 and NSF. AB - A recently described form of synaptic plasticity results in dynamic changes in the calcium permeability of synaptic AMPA receptors. Since the AMPA receptor GluR2 subunit confers calcium permeability, this plasticity is thought to occur through the dynamic exchange of synaptic GluR2-lacking and GluR2-containing receptors. To investigate the molecular mechanisms underlying this calcium permeable AMPA receptor plasticity (CARP), we examined whether AMPA receptor exchange was mediated by subunit-specific protein-protein interactions. We found that two GluR2-interacting proteins, the PDZ domain-containing Protein interacting with C kinase (PICK1) and N-ethylmaleimide sensitive fusion protein (NSF), are specifically required for CARP. Furthermore, PICK1, but not NSF, regulates the formation of extrasynaptic plasma membrane pools of GluR2 containing receptors that may be laterally mobilized into synapses during CARP. These results demonstrate that PICK1 and NSF dynamically regulate the synaptic delivery of GluR2-containing receptors during CARP and thus regulate the calcium permeability of AMPA receptors at excitatory synapses. PMID- 15797552 TI - Timing and specificity of feed-forward inhibition within the LGN. AB - Local interneurons provide feed-forward inhibition from retinal ganglion cells (RGCs) to thalamocortical (TC) neurons, but questions remain regarding the timing, magnitude, and functions of this inhibition. Here, we identify two types of inhibition that are suited to play distinctive roles. We recorded excitatory and inhibitory postsynaptic currents (EPSCs/IPSCs) in TC neurons in mouse brain slices and activated individual RGC inputs. In 34% of TC neurons, we identified EPSCs and IPSCs with identical thresholds that were tightly correlated, indicating activation by the same RGC. Such "locked" IPSCs occurred 1 ms after EPSC onset. The remaining neurons had only "nonlocked" inhibition, in which EPSCs and IPSCs had different thresholds, indicating activation by different RGCs. Nonlocked inhibition may refine receptive fields within the LGN by providing surround inhibition. In contrast, dynamic-clamp recordings suggest that locked inhibition improves the precision of synaptically evoked responses in individual TC neurons by eliminating secondary spikes. PMID- 15797554 TI - Functional imaging reveals rapid development of visual response properties in the zebrafish tectum. AB - The visual pathway from the retina to the optic tectum in fish and frogs has long been studied as a model for neural circuit formation. Although morphological aspects, such as axonal and dendritic arborization, have been well characterized, less is known about how this translates into functional properties of tectal neurons during development. We developed a system to provide controlled visual stimuli to larval zebrafish, while performing two-photon imaging of tectal neurons loaded with a fluorescent calcium indicator, allowing us to determine visual response properties in intact fish. In relatively mature larvae, we describe receptive field sizes, visual topography, and direction and size selectivity. We also characterize the onset and development of visual responses, beginning when retinal axons first arborize in the tectum. Surprisingly, most of these properties are established soon after dendrite growth and synaptogenesis begin and do not require patterned visual experience or a protracted period of refinement. PMID- 15797553 TI - Selective GABAergic control of higher-order thalamic relays. AB - GABAergic signaling is central to the function of the thalamus and has been traditionally attributed primarily to the nucleus reticularis thalami (nRT). Here we present a GABAergic pathway, distinct from the nRT, that exerts a powerful inhibitory effect selectively in higher-order thalamic relays of the rat. Axons originating in the anterior pretectal nucleus (APT) innervated the proximal dendrites of relay cells via large GABAergic terminals with multiple release sites. Stimulation of the APT in an in vitro slice preparation revealed a GABA(A) receptor-mediated, monosynaptic IPSC in relay cells. Activation of presumed single APT fibers induced rebound burst firing in relay cells. Different APT neurons recorded in vivo displayed fast bursting, tonic, or rhythmic firing. Our data suggest that selective extrareticular GABAergic control of relay cell activity will result in effective, state-dependent gating of thalamocortical information transfer in higher-order but not in first-order relays. PMID- 15797555 TI - Increased noise level of purkinje cell activities minimizes impact of their modulation during sensorimotor control. AB - While firing rate is well established as a relevant parameter for encoding information exchanged between neurons, the significance of other parameters is more conjectural. Here, we show that regularity of neuronal spike activities affects sensorimotor processing in tottering mutants, which suffer from a mutation in P/Q-type voltage-gated calcium channels. While the modulation amplitude of the simple spike firing rate of their floccular Purkinje cells during optokinetic stimulation is indistinguishable from that of wild-types, the regularity of their firing is markedly disrupted. The gain and phase values of tottering's compensatory eye movements are indistinguishable from those of flocculectomized wild-types or from totterings with the flocculus treated with P/Q-type calcium channel blockers. Moreover, normal eye movements can be evoked in tottering when the flocculus is electrically stimulated with regular spike trains mimicking the firing pattern of normal simple spikes. This study demonstrates the importance of regularity of firing in Purkinje cells for neuronal information processing. PMID- 15797556 TI - Anticipatory activity in primary motor cortex codes memorized movement sequences. AB - Movement sequences, defined both by the component movements and by the serial order in which they are produced, are fundamental building blocks of motor behavior. The serial order of sequence production is strongly encoded in medial motor areas. It is not known to what extent sequences are further elaborated or encoded in primary motor cortex. Here, we describe cells in the primary motor cortex of the monkey that show anticipatory activity exclusively related to a specific memorized sequence of upcoming movements. In addition, the injection of muscimol, a GABA agonist, into motor cortex resulted in an increase in the error rate during sequence production, without concomitant effects on nonsequenced motor performance. Our results challenge the role of medial motor areas in the control of well-practiced movement sequences and suggest that motor cortex contains a complete apparatus for the planning and production of this complex behavior. PMID- 15797558 TI - Simultaneous determination of free and N-acetylated polyamines in urine by semimicro high-performance liquid chromatography using 4-(5,6-dimethoxy-2 phthalimidinyl)-2-methoxyphenylsulfonyl chloride as a fluorescent labeling reagent. AB - We have developed a simple and highly sensitive semimicro high-performance liquid chromatographic method for the simultaneous determination of free and N acetylated polyamines in urine. Polyamines and N-acetylated polyamines were derivatized with 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride to produce fluorescent sulfonamides. The labeling reaction was carried out at 50 degrees C for 15 min at pH 9. The fluorescent derivatives were separated on a reversed-phase column with a gradient elution using water acetonitrile-methanol at 50 degrees C and detected by fluorescence measurement at 318 nm (excitation) and 406 nm (emission). The detection limits (signal-to-noise ratio=3) of the polyamines and N-acetylated polyamines were 0.7-4.5 fmol/injection. The within-day and day-to-day relative standard deviations were 3.2-7.9 and 3.0-7.7%, respectively. Significant differences were found in the urinary excretion of polyamines between cancer patients and normal subjects. PMID- 15797557 TI - Individual differences among grapheme-color synesthetes: brain-behavior correlations. AB - Grapheme-color synesthetes experience specific colors associated with specific number or letter characters. To determine the neural locus of this condition, we compared behavioral and fMRI responses in six grapheme-color synesthetes to control subjects. In our behavioral experiments, we found that a subject's synesthetic experience can aid in texture segregation (experiment 1) and reduce the effects of crowding (experiment 2). For synesthetes, graphemes produced larger fMRI responses in color-selective area human V4 than for control subjects (experiment 3). Importantly, we found a correlation within subjects between the behavioral and fMRI results; subjects with better performance on the behavioral experiments showed larger fMRI responses in early retinotopic visual areas (V1, V2, V3, and hV4). These results suggest that grapheme-color synesthesia is the result of cross-activation between grapheme-selective and color-selective brain areas. The correlation between the behavioral and fMRI results suggests that grapheme-color synesthetes may constitute a heterogeneous group. PMID- 15797559 TI - The preparation of HEK293 alpha1A or HEK293 alpha1B cell membrane stationary phase and the chromatographic affinity study of ligands of alpha1 adrenoceptor. AB - As a novel bioaffinity chromatographic technique, cell membrane chromatography (CMC) originated in 1996. The cell membrane stationary phase (CMSP) consists of porous silica coated with active cell membranes. By immersing silica into a suspension of cell membranes, the whole surface of silica was covered by the cell membranes. The present study repeatedly investigated the interaction between ligands and receptors by employing the system of CMC and especially evaluated the accuracy and feasibility of the CMC model in the study of subtype receptors. The cDNA encoding alpha1A or alpha1B adrenergic receptors (ARs) was transfected into human embryonic kidney 293 (HEK293) cell lines; cell lines stably overexpressing subtype receptors were obtained. HEK293 alpha1A or HEK293 alpha1B ARs CMSP were prepared by immobilizing relevant cell membranes on silica. In the described chromatography-based CMSP, the retention times of nine alpha1 adrenoceptor ligands and calculated capacity factors, as chromatographic parameters, were recorded carefully. These results showed a good correlation with the affinity of the same compounds for the corresponding cloned alpha1 adrenoceptor subtype. The rank order of capacity factors was consistent with the affinity rank order obtained from radioligand binding assays. The immobilized subtype-selective CMSP was stable and reproducible. The study demonstrates that the HEK293 alpha1A and HEK293 alpha1B CMSP can be utilized for initial screening of drug candidates. PMID- 15797560 TI - Direct detection of the binding of avidin and lactoferrin fluorescent probes to heparinized surfaces. AB - We describe the use of two heparin-binding proteins, avidin and lactoferrin, as probes for monitoring the amount of heparin immobilized to plastic surfaces. The proteins were derivatized with either fluorescent labels or europium chelates, enabling sensitive, fast, reproducible, and robust assays, and were used to measure the amount of protein bound to heparinized microplates, with particular attention to plates that have been coated with bovine serum albumin (BSA)-heparin conjugate. This direct method unequivocally shows that BSA-heparin affords an economical, convenient, and reliable method for coating both polystyrene microtiter plates and magnetic beads with heparin. We demonstrate that assays using directly labeled proteins overcome the problems of dissociation of the heparin-protein complex, which can occur during incubation and washing steps associated with antibody-based detection methods, and the loss in binding capacity caused by certain blocking regimes. We suggest that labeled avidin and lactoferrin are convenient probes for heparinized surfaces with the potential for much wider applicability than that presented here. PMID- 15797561 TI - Classification of the mode of inhibition of high-affinity choline uptake using capillary electrophoresis with electrochemical detection at an enzyme-modified microelectrode. AB - A nonradiochemical in vitro assay using capillary electrophoresis with electrochemical detection at an enzyme-modified microelectrode has been developed to evaluate the inhibition of high-affinity choline transport in synaptosomes. Quantitative analysis of high-affinity choline transporter rates as a function of inhibitor and substrate concentrations allowed determination of the mode of inhibition for the quaternary ammonium-catechol-based inhibitors 3 [(trimethylammonio)methyl]catechol, N,N-dimethylepinephrine, and 6-hydroxy-N,N dimethylepinephrine. The results are compared to the well-characterized inhibitor of choline transport, hemicholinium-3. PMID- 15797562 TI - Assay for the quantification of small-sized fragmented genomic DNA. AB - This paper presents a simple assay for the direct quantification of small-sized (up to 1000 bp) fragmented DNA based on the differential separation between large and small-sized DNA by polyethylene glycol precipitation. The assay can be applied to as low as 1 microg ml(-1) DNA and has a very low DNA detection limit (0.01 microg ml(-1) fragmented DNA), which can be further lowered at least 10 fold with anion exchange chromatography. The assay quantifies for the first time a more direct indicator of the extent of DNA damage than the usually assessed DNA nicks and base modifications, since small-sized fragmented DNA represents an unrepairable DNA damage of necrotic and apoptotic events that are related to normal and abnormal biological conditions. PMID- 15797563 TI - Development and evaluation of real competitive PCR for high-throughput quantitative applications. AB - Real competitive PCR (rcPCR) has been shown to have high sensitivity, reproducibility, and high-throughput potential. We describe further development and evaluation of this methodology as a tool for measuring nucleic acid abundance within a cell. Modifications to the original protocol allow analysis of gene expression levels using standard conditions regardless of mRNA abundance and assay type, thereby increasing throughput and ease of reaction setup while decreasing optimization time. In addition, we have developed a software package, TITAN, to automatically analyze the results. The details are relevant to researchers performing competitive PCR using any detection technique. The effectiveness of the described developments is demonstrated using 12 genes known to have differential expression in cell lines grown under normal and hypoxic conditions. Quantitative and qualitative comparisons to real-time PCR are presented. It is also demonstrated that the technique is capable of detecting submicroscopic chromosomal DNA deletions. PMID- 15797564 TI - Acute biochemical effects of La(NO3)3 on liver and kidney tissues by magic-angle spinning 1H nuclear magnetic resonance spectroscopy and pattern recognition. AB - High-resolution magic-angle spinning (MAS) 1H nuclear magnetic resonance (NMR) spectroscopic and pattern recognition (PR) based methods have been applied to studies on the acute biochemical effects of La(NO3)3 on rats. Male Wistar rats were treated with various doses of La(NO3)3 (2, 10, and 50 mg/kg body weight), and MAS 1H NMR spectra of their intact liver and kidney tissues were analyzed using principal components analysis to extract metabolic information. The biochemical effects of La(NO3)3 were characterized by the increase of triglyceride and bile acid and the decrease of glycogen in liver tissue, together with a slight elevation of triglyceride level in kidney tissue. The target lesion of La(NO3)3 to liver was found by MAS NMR-PR methods. This study illustrated the power of the combination of MAS 1H NMR and pattern recognition for the analysis of biochemical effects of rare earths. PMID- 15797565 TI - Alkaline phosphatase vs luciferase as secreted reporter molecules in vivo. AB - Secreted alkaline phosphatase (SEAP) and Metridia luciferase (MLuc) are useful reporter molecules in vitro, but little is understood about their usefulness in vivo. In this study, we investigated in vivo activity of recombinant SEAP and MLuc in blood and urine. When SEAP-transfected cells or recombinant SEAP were injected into rats, substantial increase in the level of serum SEAP was observed. In contrast, activity of SEAP was not detected in urine of rats injected with either the SEAP-transfected cells or recombinant SEAP. SEAP activity was also undetectable in urine of SEAP-injected Nagase analbuminemic rats in which glomerular permeability to macromolecules is enhanced. When MLuc-transfected cells were implanted into rats, activity of MLuc was undetectable not only in urine but also in serum. Even immediately after intravenous injection of recombinant MLuc, activity of MLuc was not detected in serum. Subsequent experiments revealed that, in contrast to SEAP, MLuc was rapidly inactivated either by rat serum, fetal bovine serum, or human serum. Albumin was identified as the molecule responsible for the inhibition of MLuc activity. These data elucidated advantages and limitations of secreted reporter molecules SEAP and MLuc under in vivo situations. PMID- 15797566 TI - Major artifacts encountered in studying biological samples containing ferric protoporphyrin IX. AB - Heme (ferric protoporphyrin IX, FP) dissolves very rapidly into the lipid phase of membranes, and a large number of studies have focused attention on its possible toxic effect in whole cells or isolated membranes. However, because of its molecular structure and reactivity, different problems can be encountered during the course of studying biological samples containing FP. In this article, we discuss important interferences by FP and artifacts that can affect the experimental values. First, FP interferes with the Lowry's protein determination; therefore, membranes containing FP are overestimated in their protein content determined by this procedure. Second, freezing membranes at -20 degrees C artifactually increases the local concentration of FP, thereby enhancing FP induced lipid peroxidation. Third, in the presence of thiol compounds such as N acetyl cysteine, FP is degraded to products that interfere with the thiobarbituric acid assay, one of the most widely used methods to measure the extent of lipoperoxidation. PMID- 15797567 TI - Antibody microarrays for native toxin detection. AB - We have developed antibody-based microarray techniques for the multiplexed detection of cholera toxin beta-subunit, diphtheria toxin, anthrax lethal factor and protective antigen, Staphylococcus aureus enterotoxin B, and tetanus toxin C fragment in spiked samples. Two detection schemes were investigated: (i) a direct assay in which fluorescently labeled toxins were captured directly by the antibody array and (ii) a competition assay that employed unlabeled toxins as reporters for the quantification of native toxin in solution. In the direct assay, fluorescence measured at each array element is correlated with labeled toxin concentration to yield baseline binding information (Langmuir isotherms and affinity constants). Extending from the direct assay, the competition assay yields information on the presence, identity, and concentration of toxins. A significant advantage of the competition assay over reported profiling assays is the minimal sample preparation required prior to analysis because the competition assay obviates the need to fluorescently label native proteins in the sample of interest. Sigmoidal calibration curves and detection limits were established for both assay formats. Although the sensitivity of the direct assay is superior to that of the competition assay, detection limits for unmodified toxins in the competition assay are comparable to values reported previously for sandwich format immunoassays of antibodies arrayed on planar substrates. As a demonstration of the potential of the competition assay for unlabeled toxin detection, we conclude with a straightforward multiplexed assay for the differentiation and identification of both native S. aureus enterotoxin B and tetanus toxin C fragment in spiked dilute serum samples. PMID- 15797568 TI - Solubilization, stabilization, and purification of chemokine receptors using biosensor technology. AB - Establishing solubilization conditions for membrane-associated receptors is often a tedious empirical process. Here we describe a novel application of SPR biosensor technology to screen solubilization conditions automatically and to assess receptor activity directly. We focus on two chemokine receptors, CXCR4 and CCR5, which are important in HIV cell invasion. The autosampler in Biacore 3000 permitted whole cells expressing C-terminally tagged receptors to be automatically lysed under a given solubilization condition and the lysates to be injected over an antibody surface. The total amount of solubilized receptor could be quantitated from the antibody capture level, whereas the amount of active receptor could be quantitated using a subsequent injection of conformationally sensitive antibody or protein. Using this approach, we identified detergent/lipid/buffer combinations that enhanced and maintained receptor activity. We also used the biosensor to demonstrate CD4-dependent binding of gp120 to solubilized CCR5 and to develop affinity chromatography-based purification methods that increased receptor activity more than 300%. Together, these results illustrate the benefits of using the biosensor as a tool for isolating functional membrane receptors and for analyzing ligand/receptor interactions. PMID- 15797570 TI - Touchdown thermocycling program enables a robust single nucleotide polymorphism typing method based on allele-specific real-time polymerase chain reaction. AB - Different methods have been developed for single nucleotide polymorphism (SNP) typing during recent years. Allele-specific polymerase chain reaction (ASPCR) is a cost-saving method that scores SNPs by difference of the PCR efficiency of allele-specific primers. However, ASPCR for SNP typing is notoriously confounded for its locus-specific unpredictability and the laborious gel electrophoresis. In the current study, we investigated the real-time kinetics of ASPCR and found that a simple touchdown thermocycling protocol improved its specificity significantly. Combined with real-time PCR, we developed a homogeneous genotyping method and scored more than 1000 genotypes, including all transition and transversion SNPs. A clear genotyping result was identified and validated the robustness of the method. Optimization of reactions and intrinsic modification of allele-specific primers, a laborious process but one that is repeatedly reported to be inevitable for successful ASPCR, was proved to be unnecessary with our method. Accuracy was confirmed with mass spectrometry. These characters enabled real-time ASPCR with the touchdown thermocycling protocol being very competitive among various SNP typing methods for large-scale genetic studies. PMID- 15797569 TI - Rapid sodium cyanide depletion in cell culture media: outgassing of hydrogen cyanide at physiological pH. AB - During the course of in vitro studies on cyanide exposure with SH-SY5Y human neuroblastoma cells, we found that sodium cyanide (NaCN) up to a concentration of 10 mM had no significant toxic effect under our culture conditions. Further investigation of this apparent cyanide resistance revealed that the sodium cyanide was being rapidly depleted from the cell culture medium. Cyanide was interacting with constituents of the cell culture medium and was somehow being detoxified or removed from solution. The reaction of cyanide with cell culture media in 96-well culture plates reduced cyanide concentrations rapidly (80-90% in 2 h at 37 degrees C). Running the same reaction in capped tubes significantly reduced cyanide loss from solution. Incubation of cyanide with individual constituents of the cell culture medium in solution showed that glucose, phenol red, and amino acids all acted to detoxify or remove cyanide from solution. When amino acids or buffers were incubated with sodium cyanide in aqueous solution at pH 7.4, hydrogen cyanide (HCN) was found to degas from the solutions. We compared HCN outgassing over a range of pH values. As expected, HCN remained very soluble at high pH, but as the pH was reduced to 7.0, the rate of HCN formation and outgassing increased dramatically. Acid-base reactions involving cyanide and proton donors, such as amino acids and other cell culture media constituents, at physiological pH result in rapid HCN outgassing from solution at 37 degrees C. These results indicate that previous in vitro cyanide toxicity studies done in standard culture media with prolonged incubation times using gas-exchanging culture containers might have to be reevaluated in light of the fact that the effective cyanide concentrations in the culture media were significantly lower than reported. PMID- 15797571 TI - Achiral-chiral LC/LC-MS/MS coupling for determination of chiral discrimination effects in phenprocoumon metabolism. AB - Many physiological processes show a high degree of stereoselectivity, including the metabolism of xenobiotics as catalyzed by cytochrome P450 enzymes. An analysis of these chiral discrimination effects in drug metabolism is essential for an in-depth understanding of metabolic pathways that differ between enantiomers of a given chiral drug or metabolite thereof. Achiral chromatographic separation and structural identification followed by chiral analysis of metabolites from blood specimens usually requires a time-consuming multistage analytical technique. In an effort to optimize such a complicated analytical scheme, a novel two-dimensional online achiral-chiral liquid chromatography tandem mass spectrometry (LC/LC-MS/MS) coupling method was developed by using a peak parking technique in combination with a makeup flow system. Metabolites were separated in the first dimension using a C18 reversed-phase system. A makeup eluent of water/methanol (95/5) was split into the flow before storing the metabolites separately on chiral cartridges. Subsequently, the metabolite enantiomers were eluted backward onto the analytical chiral column and separated, and the ratio of enantiomers was determined. The method was successfully validated with respect to limit of detection, linearity, intra- and interday accuracy, and precision. In the course of a human volunteer study investigating the influence of CYP (cytochrome) 2C9 genetic polymorphism on phenprocoumon (PPC) metabolism, we used this new two-dimensional online analytical technique for the analysis of PPC metabolites in plasma. The enantiomeric forms of 4'-, 6-, and 7 hydroxy-PPC metabolites as well as two novel metabolites were identified, and the ratio of the enantiomers was calculated. We found that the enantiomeric ratio for the different metabolites in the plasma sample of each measured individual differs markedly from a nearly 100% chiral discrimination for the two new putative metabolites. This new analytical coupling method possesses general utility in the analysis of chiral discrimination effects, particularly as it relates to pharmacokinetics and dynamics, a scientific field that is rapidly becoming an area of concern and interest. PMID- 15797573 TI - Capillary electrophoresis for the investigation of prostate-specific antigen heterogeneity. AB - Prostate-specific antigen (PSA) is a single-chain glycoprotein that is used as a biomarker for prostate-related diseases. PSA has one known posttranslational modification, a sialylated diantennary N-linked oligosaccharide attached to the asparagine residue N45. In this study capillary electrophoresis (CE) was employed to separate the isoforms of seven commercially available free PSA samples, two of which were specialized: enzymatically active PSA and noncomplexing PSA. The free PSA samples examined migrated as four to nine distinct, highly resolved peaks, indicating the presence of several isoforms differing in their oligosaccharide compositions. Overall, the use of CE provides a rapid, reproducible method for separation of PSA into its individual isoforms. PMID- 15797572 TI - Tandem mass spectrometric assay of succinylacetone in urine for the diagnosis of hepatorenal tyrosinemia. AB - We describe an isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of succinylacetone (SA) in urine for the diagnosis of hepatorenal tyrosinemia (HT1). The method used 15N-labeled 5(3) methyl-3(5)-isoxazole propionic acid as internal standard. Urine samples were oximated with hydroxylamine hydrochloride at 80 degrees C, extracted by solvent solvent extraction, and followed by derivatization of the butyl ester. The butylated isoxazole derivatives of SA and its internal standard were detected and quantified using positive ion electrospray LC-MS/MS with selected reaction monitoring. The turnaround time between injections was 10 min. Calibration curves were linear over the range of 0.0633-63.3 micromol/L. The intra- and interday assay variations were less than 7%. Mean recoveries of SA at three different concentrations ranged from 96 to 109%. During the course of this study, we identified 12 new patients with HT1 and applied this method to follow up the treatment of 4 of these patients as well as previously diagnosed HT1 patients. PMID- 15797574 TI - Point mutation detection with the sandwich method employing hydrogel nanospheres by the surface plasmon resonance imaging technique. AB - We propose a surface modification procedure to construct DNA arrays for use in surface plasmon resonance (SPR) imaging studies for the highly sensitive detection of a K-ras point mutation, enhanced with hydrogel nanospheres. A homobifunctional alkane dithiol was adsorbed on Au film to obtain the thiol surface, and ethyleneglycol diglycidylether (EGDE) was reacted to insert the ethyleneglycol moiety, which can suppress nonspecific adsorption during SPR analysis. Then streptavidin (SA) was immobilized on EGDE using tosyl chloride activation. Biotinylated DNA ligands were bound to the SA surface via biotin-SA interaction to fabricate DNA arrays. In SPR analysis, the DNA analyte was exposed on the DNA array and hybridized with the immobilized DNA probes. Subsequently, the hydrogel nanospheres conjugated with DNA probes were bound to the DNA analytes in a sandwich configuration. The DNA-carrying nanospheres led to SPR signal enhancement and enabled us to discriminate a K-ras point mutation in the SPR difference image. The application of DNA-carrying hydrogel nanospheres for SPR imaging assays was a promising technique for high throughput and precise detection of point mutations. PMID- 15797575 TI - Spectrophotometric and fluorometric assay of superoxide ion using 4-chloro-7 nitrobenzo-2-oxa-1,3-diazole. AB - Two highly sensitive spectrophotometric methods are developed and described for the measurement of superoxide ion radical derived from KO2 as well as O2*- generated either from the xanthine-xanthine oxidase reaction or by the addition of nicotinamide adenine dinucleotide (NADH) to skeletal muscle sarcoplasmic reticulum (SR) vesicles. These methods allow quantification of superoxide ion concentration by monitoring its reaction with 4-chloro-7-nitrobenzo-2-oxa-1,3 diazole (NBD-Cl), either by recording absorbance of the final reaction product at a wavelength of 470 nm or by measuring its fluorescence emission intensity at 550 nm using an excitation wavelength of 470 nm. The extinction coefficient of the active product was determined to be 4000 M(-1) cm(-1). A lower limit second-order bimolecular rate constant of 1.5+/-0.3x10(5) M(-1) s(-1) was estimated from kinetic stopped-flow analysis for the reaction between NBD-Cl and KO2. A plot of absorbance versus concentration of superoxide was linear over the range 2 to 200 microM KO2, whereas higher sensitivities were obtained from fluorometric measurements down into sub-micromolar concentrations with a limit of detection of 100 nM KO2. This new spectrophotometric assay showed higher specificity when compared with some other commonly used methods for detection of superoxide (e.g., nitroblue tetrazolium). Results presented showed good experimental agreement with rates obtained for the measurement of superoxide ion when compared with other well-known probes such as acetylated ferri cytochrome c and 2,3-bis(2-methoxy-4 nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT). A detailed discussion of the advantages and limitations of this new superoxide ion probe is presented. PMID- 15797576 TI - Self-priming arrest by modified random oligonucleotides facilitates the quality control of whole genome amplification. PMID- 15797577 TI - Accurate size gauging of ExoIII/S1-generated deletions by PCR analysis of ligation mixtures. PMID- 15797578 TI - Recovery of single-band proteins from polyacrylamide gels by using electrophoresis in solutions with different ionic strengths and specific weights. PMID- 15797579 TI - Fluorometric assay for bacterial primases. PMID- 15797581 TI - Strain energy density as a rupture criterion for the kidney: impact tests on porcine organs, finite element simulation, and a baseline comparison between human and porcine tissues. AB - High-velocity (up to 25 m/s) impact tests were performed on pig kidneys to characterize failure behavior at deformation rates associated with traumatic injury. Cylindrical tissue samples (n = 45) and whole perfused organs (n = 34) were impacted using both falling weights and a high-velocity pneumatic projectile impactor. Impact energy was incrementally increased until visible rupture occurred. The strain energy density failure threshold fell between 25 and 60 kJ/m3 for excised porcine tissue samples, and between 15 and 30 kJ/m3 for whole, perfused organs. The relationship between localized failure in whole organ impacts and tissue level failure thresholds observed in cylindrical tissue samples was explored using a detailed finite element model of the human kidney. The model showed good correlation between experimentally observed injury patterns and predicted strain energy density distributions within the renal parenchyma. Finally, to facilitate interpretation of the porcine renal impact results with regard to human trauma, quasi-static compression test results of freshly excised human kidney cortex samples (n = 30) were compared against similar tests on pig kidneys. Human tissues failed at Lagrange strain levels similar to porcine tissue (63+/-6.3%), but at 52% lower Lagrange stress (116+/-28 kPa), and 35% lower strain energy density (17.1+/-4.4 kJ/m3). Thus conservative interpretation of porcine test results is recommended. PMID- 15797580 TI - An equation to calculate individual muscle contributions to joint stability. AB - The purpose of the current paper was to use the energy approach to develop a simplified equation for quantifying individual muscle contributions to mechanical stability about all three axes of a particular joint. Specific examples are provided for muscles acting about the lumbar spine's L4/L5 joint. The stability equation requires input of: (1) origin and insertion coordinates, relative to the joint of interest, (2) muscle force, and (3) muscle stiffness. The muscle force must be derived from a biomechanical analysis that first results in static equilibrium about all axes being studied. The equation can also accommodate muscles with nodes that change the line of action, with respect to a particular joint, as it passes from the origin to insertion. The results from this equation were compared to those from a Moment approach using more than two million simulated muscles with three-dimensional orientations. The differences between approaches were negligible in all cases. The primary advantage of the current method is that it is very easy to implement into any 2D or 3D biomechanical model of any joint, or system of joints. Furthermore, this approach will be useful in dissecting total joint stability into the individual contributions of each muscle for various systems, joints, postures and recruitment patterns. PMID- 15797582 TI - Effects on the crank torque profile when changing pedalling cadence in level ground and uphill road cycling. AB - Despite the importance of uphill cycling performance during cycling competitions, there is very little research investigating uphill cycling, particularly concerning field studies. The lack of research is partly due to the difficulties in obtaining data in the field. The aim of this study was to analyse the crank torque in road cycling on level and uphill using different pedalling cadences in the seated position. Seven male cyclists performed four tests in the seated position (1) on level ground at 80 and 100 rpm, and (2) on uphill road cycling (9.25% grade) at 60 and 80 rpm.The cyclists exercised for 1 min at their maximal aerobic power. The bicycle was equipped with the SRM Training System (Schoberer, Germany) for the measurement of power output (W), torque (Nm), pedalling cadence (rpm), and cycling velocity (km h(-1)). The most important finding of this study indicated that at maximal aerobic power the crank torque profile (relationship between torque and crank angle) varied substantially according to the pedalling cadence and with a minor effect according to the terrain. At the same power output and pedalling cadence (80 rpm) the torque at a 45 degrees crank angle tended (p < 0.06) to be higher (+26%) during uphill cycling compared to level cycling. During uphill cycling at 60 rpm the peak torque was increased by 42% compared with level ground cycling at 100 rpm. When the pedalling cadence was modified, most of the variations in the crank torque profile were localised in the power output sector (45 degrees to 135 degrees). PMID- 15797583 TI - Strain-rate dependent material properties of the porcine and human kidney capsule. AB - This study was performed to characterize the mechanical properties of the kidney capsular membrane at strain-rates associated with blunt abdominal trauma. Uniaxial quasi-static and dynamic tensile experiments were performed on fresh, unfrozen porcine and human renal capsules at deformation rates ranging from 0.0001 to 7 m/s (strain-rates of 0.005-250 s(-1)). Single stroke, dynamic tests were performed on samples of porcine renal capsule at strain-rates of 0.005 s(-1) (n = 33), 0.05 s(-1) (n = 17), 0.5 s(-1) (n = 38), 2 s(-1) (n = 10), 4 s(-1) (n = 10), 50 s(-1) (n = 21), 100 s(-1) (n = 18), 150 s(-1) (n = 17), 200 s(-1) (n = 10), and 250 s(-1) (n = 17). Due to limited availability of human tissues, only quasi-static tests were performed (0.005 s(-1), n = 25). Porcine renal capsule properties were found to match the material properties of human capsular tissue sufficiently well such that porcine tissue material can be used as a human test surrogate. The apparent elastic modulus and breaking stress of the porcine renal capsule were observed to increase significantly with increasing strain-rate (p < 0.01). Breaking strain was inversely related to strain-rate (p < 0.01). The effect of increasing strain-rate on material properties diminished appreciably at rates exceeding 150 s(-1). Empirically derived mathematical models of constitutive behavior were developed using a hyperelastic/viscoelastic Ogden formulation, as well as a Cowper-Symonds law material curve multiplication. PMID- 15797584 TI - Bone ingrowth simulation for a concept glenoid component design. AB - Glenoid component loosening is the major problem of total shoulder arthroplasty. It is possible that uncemented component may be able to achieve superior fixation relative to cemented component. One option for uncemented glenoid is to use porous tantalum backing. Bone ingrowth into the porous backing requires a degree of stability to be achieved directly post-operatively. This paper investigates the feasibility of bone ingrowth with respect to the influence of primary fixation, elastic properties of the backing and friction at the bone prosthesis interface. Finite element models of three glenoid components with different primary fixation configurations are created. Bone ingrowth into the porous backing is modelled based on the magnitude of the relative interface micromotions and mechanoregulation of the mesenchymal stem cells that migrated via the bonded part of the interface. Primary fixation had the most influence on bone ingrowth. The simulation showed that its major role was not to firmly interlock the prosthesis, but rather provide such a distribution of load, that would result in reduction of the peak interface micromotions. Should primary fixation be provided, friction has a secondary importance with respect to bone ingrowth while the influence of stiffness was counter intuitive: a less stiff backing material inhibits bone ingrowth by higher interface micromotions and stimulation of fibrous tissue formation within the backing. PMID- 15797585 TI - Effects of non-contractile inclusions on mechanical performance of skeletal muscle. AB - Glycogen storage disease II is an inherited progressive muscular disease in which the lack of functional acid 1-4 alpha-glucosidase results in the accumulation of lysosomal glycogen. In the present study, we examine the effect of these non contractile inclusions on the mechanical performance of skeletal muscle. To this end, force developed in an isometrically contracting slice of a muscle was calculated with a finite element model. Force was calculated at several inclusion densities and distributions and compared to muscle lacking inclusions. Furthermore, ankle dorsal flexor torque was measured in situ of alpha-glucosidase null mice of 6 months of age and unaffected litter mates as was inclusion density in the dorsal flexor muscles. The calculated force loss was shown to be almost exclusively dependent on the inclusion density and less on the type of inclusion distribution. The force loss predicted by the model (6%) on the basis of measured inclusion density (3.3%) corresponded to the loss in mass-normalized strength in these mice measured in situ (7%). Therefore, we conclude that the mechanical interaction between the non-contractile inclusions and the nearby myofibrils is a key factor in the loss of force per unit muscle mass during early stages of GSD II in mice. As glycogen accumulation reaches higher levels in humans, it is highly probable that the impact of this mechanical interaction is even more severe in human skeletal muscle. PMID- 15797586 TI - Three-dimensional finite element analysis of the foot during standing--a material sensitivity study. AB - Information on the internal stresses/strains in the human foot and the pressure distribution at the plantar support interface under loading is useful in enhancing knowledge on the biomechanics of the ankle-foot complex. While techniques for plantar pressure measurements are well established, direct measurement of the internal stresses/strains is difficult. A three-dimensional (3D) finite element model of the human foot and ankle was developed using the actual geometry of the foot skeleton and soft tissues, which were obtained from 3D reconstruction of MR images. Except the phalanges that were fused, the interaction among the metatarsals, cuneiforms, cuboid, navicular, talus, calcaneus, tibia and fibula were defined as contact surfaces, which allow relative articulating movement. The plantar fascia and 72 major ligaments were simulated using tension-only truss elements by connecting the corresponding attachment points on the bone surfaces. The bony and ligamentous structures were embedded in a volume of soft tissues. The encapsulated soft tissue was defined as hyperelastic, while the bony and ligamentous structures were assumed to be linearly elastic. The effects of soft tissue stiffening on the stress distribution of the plantar surface and bony structures during balanced standing were investigated. Increases of soft tissue stiffness from 2 and up to 5 times the normal values were used to approximate the pathologically stiffened tissue behaviour with increasing stages of diabetic neuropathy. The results showed that a five-fold increase in soft tissue stiffness led to about 35% and 33% increase in the peak plantar pressure at the forefoot and rearfoot regions, respectively. This corresponded to about 47% decrease in the total contact area between the plantar foot and the horizontal support surface. Peak bone stress was found at the third metatarsal in all calculated cases with a minimal increase of about 7% with soft tissue stiffening. PMID- 15797587 TI - Individual flight styles in ski jumping: results obtained during Olympic Games competitions. AB - From the physics point of view, the jump length in ski jumping depends on: the in run velocity v(0), the velocity perpendicular to the ramp v(p0) due to the athlete's jumping force, the lift and drag forces acting during take-off and during the flight, and the weight of the athlete and his equipment. The aerodynamic forces are a function of the flight position and of the equipment features. They are a predominant performance factor and can largely be influenced by the athlete. The field study conducted during the Olympic Games competitions 2002 at Park City (elevation: 2000 m) showed an impressive ability of the Olympic medallists to reproduce their flight style and remarkable differences between different athletes have been found. The aerodynamic forces are proportional to the air density. Elite athletes are able to adapt their flight style to thin air conditions in order to maximise jump length and to keep the flight stable. The effects of flight position variations on the performance have been analysed by means of a computer model which is based on the equations of motion and on wind tunnel data corresponding to the flight positions found in the field. Athletes have to solve extremely difficult optimisation problems within fractions of a second. The computer simulation can be used as a reliable starting point for the improvement of training methods and gives an insight into the "implicit" knowledge of physics that the ski jumping athlete must have available for a good performance. PMID- 15797588 TI - Structural properties of the medial collateral ligament complex of the human knee. AB - The anatomy of the medial collateral ligament (MCL) complex consists of three identifiable passive restraining structures: the longitudinal fibres of the superficial medial collateral ligament (sMCL), the deep medial collateral ligament (dMCL), and the posteromedial capsule (PMC). The purpose of this study was to measure and compare the structural properties of these three individual structures. Eight human cadaveric knees (age 72-89 years, mean = 77 years, S.D. 5.3) were harvested and bone-ligament-bone tensile testing specimens prepared. After preconditioning, the specimens were extended to failure at 1000 mm/min in an Instron tensile testing machine. Ligament bundles failed either mid-substance or at their bony attachments. The ligament bundles had maximum loads of 534 N (sMCL), 194 N (dMCL), 425 N (PMC) and failed at 10.2, 7.1, and 12.0 mm mean extension, respectively. The maximum load and linear stiffness of the sMCL were significantly higher than those of the dMCL but not the PMC. The maximum load of the PMC was significantly higher than that of the dMCL; the linear stiffness of the PMC was higher than that of the dMCL but this did not reach statistical significance. The dMCL failed at a significantly lower extension than the other structures. The sMCL bundles that failed at their bony attachment were remounted using a freezing clamp fixture and again extended to failure, resulting in mid substance failure at 884 N (74% higher). This study has shown that the PMC of the knee has comparable structural properties to the long superficial MCL and the short, deep MCL. In summary, the structural properties of the different component structures of the medial ligament complex indicate possible functional significance. PMID- 15797589 TI - Cruciate coupling and screw-home mechanism in passive knee joint during extension -flexion. AB - The screw-home mechanism and coupling between forces in cruciate ligaments during passive knee joint flexion were investigated for various boundary conditions, flexion axis alignments and posterior cruciate ligaments (PCL)/anterior cruciate ligament (ACL) conditions. A developed non-linear 3D finite element model was used to perform detailed elasto-static response analyses of the human tibiofemoral joint as a function of flexion angle varying from 10 degrees hyper extension to 90 degrees flexion. The tibia rotated internally as the femur flexed and externally as the femur extended. The re-alignment of the flexion axis by +/ 5 degrees rotation about the axial (distal-proximal) axis, transection of the ACL and changes in cruciate ligament initial strains substantially influenced the 'screw-home' motion. On the other hand, restraint on this coupled rotation diminished ACL forces in flexion. A remarkable coupling was predicted between ACL and PCL forces in flexion; forces in both cruciate ligaments increased as the initial strain or pretension in one of them increased whereas they both diminished as one of them was cut or became slack. This has important consequences in joint functional biomechanics following a ligament injury or replacement surgery and, hence, in the proper management of joint disorders. PMID- 15797590 TI - Application of singular spectrum analysis to the smoothing of raw kinematic signals. AB - Motion capture systems currently used in biomechanical analysis introduce systematic measurement errors that appear in the form of noise in recorded displacement signals. The noise is unacceptably amplified when differentiating displacements to obtain velocities and accelerations. To avoid this phenomenon, it is necessary to smooth the displacement signal prior to differentiation in order to eliminate the noise introduced by the experimental system. The use of singular spectrum analysis (SSA) is presented in this paper as an alternative to traditional digital filtering methods. SSA is a novel non-parametric technique based on principles of multivariate statistics. The original time series is decomposed into a number of additive time series, each of which can be easily identified as being part of the modulated signal, or as being part of the random noise. Several examples that demonstrate the superiority of this technique over other methods used in biomechanical analysis are presented in this paper. PMID- 15797592 TI - In vivo debonding strength and enamel damage in two orthodontic debonding methods. AB - Bracket debonding strength related to diverse debonding methods and enamel damage has not been assessed in vivo. The study hypothetized a direct relationship between these three parameters. Debonding strength was measured clinically in the wings method and base method on 50 patients in a split mouth method using a calibrated debonding plier. Brackets from 30 of these patients were scanned in SEM and EDAX for adhesive remnant index and enamel calcium remnants. Base method debonding force was significantly greater than wings method (163.5+/-68.7 N, 106.1+/-66.2 N, respectively, p < 0.001). A positive adhesive remnant index score was found in both methods (68.7%, 66.7%, respectively). Debonding strength vs. adhesive remnant index or calcium index scores were not correlated. However, the latter two were significantly correlated (0.524 < R < 0.895, p < 0.031). Half of the debonding failures developed at the adhesive enamel interface. The results warnts the potential of enamel damage during debonding. PMID- 15797591 TI - High rate shear strain of three-dimensional neural cell cultures: a new in vitro traumatic brain injury model. AB - The fidelity of cell culture simulations of traumatic brain injury (TBI) that yield tolerance and mechanistic information relies on both the cellular models and mechanical insult parameters. We have designed and characterized an electro mechanical cell shearing device in order to produce a controlled high strain rate injury (up to 0.50 strain, 30 s(-1) strain rate) that deforms three-dimensional (3-D) neural cultures (neurons or astrocytes in an extracellular matrix scaffold). Theoretical analysis revealed that these parameters generate a heterogeneous 3-D strain field throughout the cultures that is dependent on initial cell orientation within the matrix, resulting in various combinations of normal and shear strain. The ability to create a linear shear strain field over a range of input parameters was verified by tracking fluorescent microbeads in an acellular matrix during maximal displacement for a range of strains and strain rates. In addition, cell death was demonstrated in rat cortical astrocytes and neurons in response to high rate, high magnitude shear strain. Furthermore, cell response within the 3-D neuronal cultures depended on orientation, with higher predicted shear strain correlating with an increased loss of neurites, indicating that culture configuration may be an important factor in the mechanical, and hence cellular, response to traumatic insults. Collectively, these results suggest that differential responses exist within a 3-D culture subjected to mechanical insult, perhaps mimicking the in vivo environment, and that this new model can be used to investigate the complex cellular mechanisms associated with TBI. PMID- 15797593 TI - Modeling rough stenoses by an immersed-boundary method. AB - A pulsatile laminar flow of a viscous, incompressible fluid through a stenosed artery was simulated by an immersed-boundary method. The method allows the use of a simple (rectangular) computational domain in order to simulate a flow around a complex geometry obstacle with surface irregularities (roughness). The influence of the shape and the surface roughness on the flow resistance was explored. The obtained numerical results were validated by comparison with published experimental and numerical results. We show that the surface irregularities have no significant influence on the flow resistance across an obstacle for a physiological range of Reynolds numbers. Notwithstanding, an accurate representation of irregularities allows investigation of the near-wall effects of a realistic flow such as fluid recirculation. We show that a detailed study of flow patterns in the immediate vicinity of the irregular surface can be performed using the immersed boundary method. PMID- 15797594 TI - Multiscale modeling of the cardiovascular system: application to the study of pulmonary and coronary perfusions in the univentricular circulation. AB - The objective of this study is to compare the coronary and pulmonary blood flow dynamics resulting from two configurations of systemic-to-pulmonary artery shunts currently utilized during the Norwood procedure: the central (CS) and modified Blalock Taussig (MBTS) shunts. A lumped parameter model of the neonatal cardiovascular circulation and detailed 3-D models of the shunt based on the finite volume method were constructed. Shunt sizes of 3, 3.5 and 4 mm were considered. A multiscale approach was adopted to prescribe appropriate and realistic boundary conditions for the 3-D models of the Norwood circulation. Results showed that the average shunt flow rate is higher for the CS option than for the MBTS and that pulmonary flow increases with shunt size for both options. Cardiac output is higher for the CS option for all shunt sizes. Flow distribution between the left and the right pulmonary arteries is not completely balanced, although for the CS option the discrepancy is low (50-51% of the pulmonary flow to the right lung) while for the MBTS it is more pronounced with larger shunt sizes (51-54% to the left lung). The CS option favors perfusion to the right lung while the MBTS favors the left. In the CS option, a smaller percentage of aortic flow is distributed to the coronary circulation, while that percentage rises for the MBTS. These findings may have important implications for coronary blood flow and ventricular function. PMID- 15797595 TI - Stair climbing is more critical than walking in pre-clinical assessment of primary stability in cementless THA in vitro. AB - Pre-clinical testing of hip endoprostheses is a mandatory requirement before clinical release. Inadequate loading conditions may lead to lower elastic and plastic interface movements than those occurring post-operatively in vivo. This study investigated the influence of patient activity on the primary stability of cementless prostheses with a special emphasis on active simulation of muscle forces. A loading setup, based on validated musculo-skeletal analyses, was used to generate the hip contact force during walking and stair climbing by transmitting muscle forces through the femur. In addition, a loading configuration which only generated the hip contact force occurring during stair climbing at the prosthesis head was simulated. CLS prostheses were implanted in 18 composite femora and subjected to cyclical loading. The relative micro movements at the bone-prosthesis interface were determined and appeared to be extremely sensitive to the specific patient activity. Compared to walking, stair climbing generated higher micro-movements, with pronounced axial and rotational components. Stair climbing with the femur loaded by the resultant hip contact force only exhibited a characteristic valgus tilt of the stem with significantly lower interface micro-movements than under active simulation of muscle forces. The analyses suggest that stair climbing induced the highest mechanical instability at the bone-prosthesis interface, a level which may compromise the necessary osseointegration process. Active simulation of muscle forces considerably affects the primary stability of cementless hip endoprostheses. Pre clinical in vitro tests should therefore simulate stair climbing and include muscle activity in the assessment of initial implant stability, otherwise micro movements may be underestimated and the primary stability overestimated. PMID- 15797597 TI - Distribution of mechanical impedance at the fingers and the palm of the human hand. AB - A comprehensive understanding of the complex biodynamic response of the human fingers-hand-arm system may help researchers determine the causation of injuries arising from hand-transmitted vibration. This study theoretically demonstrates that the mechanical impedance (MI) in a hand power grip, as a measure of the biodynamic response of the system, can be divided into finger MI and palm MI. A methodology is developed to measure them separately and to investigate their distribution characteristics. This study involves 6 adult male subjects, constant velocity sinusoidal excitations at 10 different discrete frequencies (16, 25, 40, 63, 100, 160, 250, 400, 630, 1000 Hz), and three different hand-handle coupling conditions. Our results suggest that at low frequencies (40 Hz), the palm MI is substantially higher than the finger MI; the majority of the hand MI remains distributed at the palm up to 100 Hz; and at frequencies higher than 160 Hz, the finger MI is comparable to or higher than the palm MI. Furthermore, at frequencies equal to or above 100 Hz, the finger MI is practically independent of the palm-handle coupling conditions. Knowledge of the MI distribution pattern may increase the understanding of vibration transmission to the hand and aid in the development of effective isolation devices. PMID- 15797596 TI - Determination of muscle loading at the hip joint for use in pre-clinical testing. AB - The stability of joint endoprostheses depends on the loading conditions to which the implant-bone complex is exposed. Due to a lack of appropriate muscle force data, less complex loading conditions tend to be considered in vitro. The goal of this study was to develop a load profile that better simulates the in vivo loading conditions of a "typical" total hip replacement patient and considers the interdependence of muscle and joint forces. The development of the load profile was based on a computer model of the lower extremities that has been validated against in vivo data. This model was simplified by grouping functionally similar hip muscles. Muscle and joint contact forces were computed for an average data set of up to four patients throughout walking and stair climbing. The calculated hip contact forces were compared to the average of the in vivo measured forces. The final derived load profile included the forces of up to four muscles at the instances of maximum in vivo hip joint loading during both walking and stair climbing. The hip contact forces differed by less than 10% from the peak in vivo value for a "typical" patient. The derived load profile presented here is the first that is based on validated musculoskeletal analyses and seems achievable in an in vitro test set-up. It should therefore form the basis for further standardisation of pre-clinical testing by providing a more realistic approximation of physiological loading conditions. PMID- 15797598 TI - Enforced exercise after blunt trauma significantly affects biomechanical and histological changes in rabbit retro-patellar cartilage. AB - Our laboratory has developed an animal model to study factors leading to chronic disease in a blunt impacted joint. Studies to date indicate post trauma softening of the impacted joint cartilage, but a limited degree of histological degradation in the tissue. The model utilizes treadmill exercise of the animal post trauma. The hypothesis of the current study was that post trauma exercise helps limit histological and mechanical degradation of the impacted retro-patellar cartilage. The study involved a group of animals with enforced exercise on a treadmill and another group with cage-activity post trauma. The animals were sacrificed after 24 months. Mechanical and histological analyses were performed on the retro patellar cartilage from each group. The impacted versus contra-lateral, non impacted retro-patellar cartilage was mechanically softened in the exercise group, but not in the cage-activity group. Histological analyses of the tissue from the cage-activity group indicated that this cartilage had less surface integrity, more ossification/calcification, and more erosion than that in the impacted tissue from the exercise group. These tissue changes may lead to an apparent stiffening effect in the impacted cartilage from the cage-activity group at 24 months post-trauma. Potential relationships between the intensity and frequency of post trauma exercise and the mechanical character and histological degradation of the impacted cartilage need additional study. The study indicates that post-trauma exercise can significantly alter the outcome of a blunt knee joint trauma in this experimental animal model. PMID- 15797600 TI - Comment on "Nanoindentation and whole-bone bending estimates of material properties in bones from senescence accelerated mouse SAMP6". PMID- 15797599 TI - Performance of orientation sensors for use with a functional electrical stimulation mobility system. AB - The purpose of this study was to verify the performance of recently developed body-worn sensor packs against 3D motion analysis of trunk and lower-limb movements. Five sensor packs, each consisting of rate gyroscope and two 2-D accelerometers controlled by a microprocessor were attached to the trunk, thighs, and shanks of an able bodied subject. A 6-camera motion analysis system (MAS) recorded multiple trials of sit-to-stand movements and normal walking. Time domain signals from each sensor pack were significantly correlated (r = 0.90 0.99;p < 0.05) with a root mean square errors of less than 5 degrees when compared against the same limb angle measurements calculated by the MAS. These data demonstrate that these external sensor packs are accurate devices for measuring trunk and lower-limb sagittal plane orientation in real-time. PMID- 15797602 TI - Secondary metabolites of slime molds (myxomycetes). AB - The compounds reported from the slime molds (myxomycetes) species are described. Almost 100 natural compounds including their chemical structures and biological activities are described in this review article. Only metabolites with a well defined structure are included. PMID- 15797603 TI - Wax constituents on the inflorescence stems of double eceriferum mutants in Arabidopsis reveal complex gene interactions. AB - To shed new light on gene involvement in plant cuticular-wax production, 11 eceriferum (cer) mutants of Arabidopsis having dramatic alterations in wax composition of inflorescence stems were used to create 14 double cer mutants each with two homozygous recessive cer loci. A comprehensive analysis of stem waxes on these double mutants revealed unexpected CER gene interactions and new ideas about individual CER gene functions. Five of the 14 double cer mutants produced significantly more total wax than one of their respective cer parents, indicating from a genetic standpoint a partial bypassing (or complementation) of one cer mutation by the other. Eight of the 14 double cer mutants had alkane amounts lower than both respective cer parents, suggesting that most of these CER gene products play a major additive role in alkane synthesis. Other results suggested that some CER genes function in more than one step of the wax pathway, including those associated with sequential steps in acyl-CoA elongation. Surprisingly, complete epistasis was not observed for any of the cer gene combinations tested. Significant overlap or redundancy of genetic operations thus appears to be a central feature of wax metabolism. Future studies of CER gene product function, as well as the utilization of CER genes for crop improvement, must now account for the complex gene interactions described here. PMID- 15797604 TI - Lipid metabolism in arbuscular mycorrhizal roots of Medicago truncatula. AB - The peroxidation of polyunsaturated fatty acids, common to all eukaryotes, is mostly catalyzed by members of the lipoxygenase enzyme family of non-heme iron containing dioxygenases. Lipoxygenase products can be metabolized further in the oxylipin pathway by several groups of CYP74 enzymes. One prominent oxylipin is jasmonic acid (JA), a product of the 13-allene oxide synthase branch of the pathway and known as signaling substance that plays a role in vegetative and propagative plant development as well as in plant responses to wounding and pathogen attack. In barley roots, JA level increases upon colonization by arbuscular mycorrhizal fungi. Apart from this first result regarding JA, no information is available on the relevance of lipidperoxide metabolism in arbuscular mycorrhizal symbiosis. Thus we analyzed fatty acid and lipidperoxide patterns in roots of Medicago truncatula during mycorrhizal colonization. Levels of fungus-specific fatty acids as well as palmitic acid (16:0) and oleic acid (18:1 n - 9) were increased in mycorrhizal roots. Thus the degree of arbuscular mycorrhizal colonization of roots can be estimated via analysis of fungal specific esterified fatty acids. Otherwise, no significant changes were found in the profiles of esterified and free fatty acids. The 9- and 13-LOX products of linoleic and alpha-linolenic acid were present in all root samples, but did not show significant differences between mycorrhizal and non-mycorrhizal roots, except JA which showed elevated levels in mycorrhizal roots. In both types of roots levels of 13-LOX products were higher than those of 9-LOX products. In addition, three cDNAs encoding CYP74 enzymes, two 9/13-hydroperoxide lyases and a 13-allene oxide synthase, were isolated and characterized. The transcript accumulation of these three genes, however, was not increased in mycorrhizal roots of M. truncatula. PMID- 15797605 TI - Methoxylated fatty acids in Blumeria graminis conidia. AB - The total fatty acids (FA) composition of Blumeria graminis f.sp. tritici conidia, the causal agent of wheat powdery mildew, was analyzed as a function of their age. A total of 19 FA (C12-C24 saturated and unsaturated) and unusual methoxylated fatty acids (mFA) were detected in young, intermediate and old conidia. Two very long chain methoxylated FA were identified by GC-MS as 3 methoxydocosanoic and 3-methoxytetracosanoic acids. Medium chain FA were predominant in young conidia (75%, including 13% of mFA) while very long chain fatty acids constituted the major compounds in old conidia (74%, including 30% of mFA). We have shown for the first time that the total FA composition is strongly correlated with the age of B. graminis f.sp. tritici (Bgt) conidia. PMID- 15797606 TI - Cohaerins A and B, azaphilones from the fungus Hypoxylon cohaerens, and comparison of HPLC-based metabolite profiles in Hypoxylon sect. Annulata. AB - Azaphilones, named cohaerins A and B were isolated from stromata of the xylariaceous ascomycete Hypoxylon cohaerens. Their absolute structures were determined by spectroscopic methods (2D NMR, MS, IR, UV CD), and subsequently confirmed by acetylation. Stromatal metabolite profiles of several taxa of Hypoxylon sect. Annulata were also generated using analytical HPLC with diode array and MS detection. The cohaerins were neither found in other Hypoxylon spp., nor in other Xylariaceae. However, they were present even in holotype material of H. cohaerens, collected over 200 years ago. The binaphthalene BNT was also omnipresent in sect Annulata, and its derivatives, the benzo[j]fluoranthenes daldinone A and truncatone, as well as presumably related compounds. These fungi were found devoid of other types of azaphilone pigments of the Xylariaceae, such as mitorubrins and daldinins, the latter of which are widespread in certain groups of Hypoxylon sect. Hypoxylon. Hence, chemotaxonomic data largely support the current generic concept. The original source of truncatone was identified as Hypoxylon annulatum. PMID- 15797608 TI - Laetiporic acids, a family of non-carotenoid polyene pigments from fruit-bodies and liquid cultures of Laetiporus sulphureus (Polyporales, Fungi). AB - The non-isoprenoid polyene laetiporic acid A, recently described from fruit bodies of the wood-rotting fungus Laetiporus sulphureus, was found to be the major orange pigment also in mycelium grown in liquid culture. Its formation was variable, ranging from 0.1 to 6.7 mg/g dry weight in three strains, all of which were identified as L. sulphureus by ITS rDNA sequence analysis. A second pigment, 2-dehydro-3-deoxylaetiporic acid A, is also described and fully characterized by NMR spectroscopy. Two further minor pigments, laetiporic acids B and C, were produced in liquid culture. These resemble laetiporic acid A but are enlarged by two and four carbon atoms, respectively, resulting in chromophores with 11 or 12 instead of 10 conjugated double bonds as described for laetiporic acid A. Since fruit-bodies of L. sulphureus are edible, laetiporic acids might hold potential as food colourants. PMID- 15797607 TI - Cytotoxic lignans from Larrea tridentata. AB - Six lignans, including the cyclolignan 3,4'-dihydroxy-3',4'-dimethoxy-6,7' cyclolignan, were isolated from the flowering tops of Larrea tridentata. Additionally the flavanone, (S)-4',5-dihydroxy-7-methoxyflavanone, was isolated for the first time from L. tridentata or any member of the family Zygophyllaceae. All of the compounds were assessed for their growth inhibitory activity against human breast cancer, human colon cancer and human melanoma cell lines. The lignans had IC50 values of 5-60 microM with the linear butane-type lignans being the most potent, and it was found that colon cancer cells were the least sensitive cell type tested. The relative potency of linear butane type lignans against human breast cancer appears to correlate positively with the number of O methyl groups present on the molecule. PMID- 15797609 TI - Haemolytic acylated triterpenoid saponins from Harpullia austro-caledonica. AB - Eight new acylated triterpenoid saponins were isolated from the stem bark of Harpullia austro-caledonica along with the known harpuloside (9). Their structures were established using 1D and 2D NMR and mass spectrometry as 3-O-beta D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-21 beta, 22 alpha-di-O angeloylbarringtogenol C (1), 3-O-alpha-L-rhamnopyranosyl-(1-->3)-[beta-D galactopyranosyl-(1-->2)]-beta-D-glucuronopyranosyl-21 beta, 22 alpha-di-O angeloyl barringtogenol C (2), 3-O-alpha-L-arabinofuranosyl-(1-->3)-[beta-D galactopyranosyl-(1-->2)]-beta-D-glucuronopyranosyl-21 beta, 22 alpha-di-O angeloylbarringtogenol C (3), 3-O-alpha-L-arabinofuranosyl-(1-->2)-beta-D glucuronopyranosyl-21 beta, 22 alpha-di-O-angeloylprotoaescigenin (4), 3-O-alpha L-arabinofuranosyl-(1-->3)-[alpha-L-arabinofuranosyl-(1-->2)]-beta-D glucuronopyranosyl-21 beta, 22 alpha-di-O-angeloyl protoaescigenin (5), 3-O-alpha L-arabinofuranosyl-(1-->3)-[beta-D-xylopyranosyl-(1-->2)]-beta-D glucuronopyranosyl-21 beta, 22 alpha-di-O-angeloylprotoaescigenin (6), 3-O-alpha L-arabinofuranosyl-(1-->3)-[beta-D-glucopyranosyl-(1-->2)]-beta-D glucuronopyranosyl-21 beta, 22 alpha-di-O-angeloylprotoaescigenin (7), 3-O-beta-D xylopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-21 beta, 22 alpha-di-O angeloylprotoaescigenin (8). The EtOH extract of the stem bark showed in vitro cytotoxic activity against KB cells (90% at 10 microg/ml). At a concentration of 5 microg/ml, the saponin mixture showed haemolytic activity and caused 100% haemolysis of a 10% suspension of sheep erythrocytes. PMID- 15797610 TI - Norditerpene and diterpene alkaloids from Aconitum variegatum. AB - Aerial parts of Aconitum variegatum L. from the Pyrenees furnished four norditerpene alkaloids, 16 beta-hydroxycardiopetaline, 8-ethoxysachaconitine, 14 acetylgenicunine B, N-deethyl-N-19-didehydrosachaconitine, five diterpene alkaloids 15-veratroyldictizine, 15-veratroyl-17-acetyldictizine, 15-veratroyl-17 acetyl-19-oxodictizine, N-ethyl-1 alpha-hydroxy-17-veratroyldictizine, variegatine and the known alkaloids sachaconitine, 14-O-acetylsachaconitine, karakoline, talatizamine, 10-hydroxytalatizamine, 14-acetyltalatizamine, 14 acetyl-10-hydroxytalatizamine, N-methylarmepavine, pengshenin B, delsoline, dihydrodelsoline, delcosine and genicunin B. Structures of the alkaloids were established by MS, 1D- and 2D-NMR techniques. PMID- 15797611 TI - The complexity of genetic variation in a simple immune system. AB - Fruitflies derived from a wild population vary in their resistance to infection with the bacterial pathogen Serratia marcescens. A survey of nucleotide diversity in 21 genes involved in innate immunity concluded that 16 genes had polymorphisms associated with resistance to this specific pathogen. However, the effects of individual polymorphisms on the resistance phenotype were modest, and epistatic interactions appeared to be common. What might these findings tell us about genetic resistance to infection in humans? PMID- 15797612 TI - High rate of horizontal transfer of transposable elements in Drosophila. AB - We have conducted molecular population genetics analyses to understand the relationships among the transposable elements (TEs) in Drosophila melanogaster, in combination with sequence comparisons of TEs from two related species, D. simulans and D. yakuba. We observed much lower than expected genetic differences among elements, clear evidence for departure from expectations for equilibrium copy numbers and little divergence between species. This suggests that a large proportion of TEs in D. melanogaster had a recent origin as a result of interspecies movement. PMID- 15797613 TI - Human antisense genes have unusually short introns: evidence for selection for rapid transcription. AB - We suggest that small introns are favored in genes requiring a minimal response time ('nimble' genes), and that antisense genes might be prime candidates for such nimble genes. In this article, we show that antisense genes have significantly shorter introns than all other gene categories, which supports our proposal that the short introns of antisense genes might be functionally important and integrally related to their potential role in efficient gene regulation. PMID- 15797614 TI - Enhancer sequence conservation between vertebrates is favoured in developmental regulator genes. AB - Sequence conservation has been used to find genes and to pinpoint functional non coding sequences such as transcriptional regulatory elements. In this article, we analysed the conservation of 104 experimentally validated murine enhancer sequences between the mouse and zebrafish genomes. Surprisingly, only 10.5% of the mouse enhancers have homologues in zebrafish. All of the genes with conserved cis-elements have regulatory functions during embryonic development, perhaps reflecting substantial structural constraints on the integration of spatio temporal signalling cues during the formation of the vertebrate body. PMID- 15797615 TI - Modular genes with metazoan-specific domains have increased tissue specificity. AB - We have systematically examined the domain composition across a comprehensive set of tissue-specific, midrange and housekeeping genes as defined by their mode of expression in 52 normal mouse tissues. We show a definite correlation between the number of domains and the degree of tissue specificity. This trend is further supported by a novel analysis involving the time of origin of each domain. Genes containing metazoan-specific domains are more prevalent in signal transduction and cell-communication pathways, and are depleted in primary metabolism. Our analyses suggest that highly modular gene products have been recruited for tissue specific functions that are required in complex organisms. PMID- 15797616 TI - Genes of domestic mammals augmented by backcrossing with wild ancestors. AB - Both archaeological data and the presence of few mitochondrial DNA lineages suggest that most widespread domestic mammals (cattle, sheep, goats, pigs and dogs) derive from only a handful of domestication events. However, each of these species shows a high level of diversity at the nuclear genes of the major histocompatibility complex (MHC). Through simulations incorporating various degrees of population subdivision, growth rate and selection, we demonstrate that the numerous MHC DRB alleles that are present in modern domestic mammals implies that substantial backcrossing with wild ancestors, either accidental or intentional, has been important in shaping the genetic diversity of our domesticates. These results support the view that, contrary to common assumption, domestic and wild lineages might not have been clearly separated throughout their history. PMID- 15797617 TI - Dosage balance in gene regulation: biological implications. AB - Classical studies in genetics involving aneuploidy and ploidy comparisons and sex determination mechanisms indicated a balance phenomenon such that changes of individual chromosomal dosage altered the phenotype more dramatically than changes in ploidy. Recent evidence suggests that a major contributor to this balance is the behavior of molecular complexes that function in various regulatory processes affecting gene expression. In this article, we discuss the potential contribution of regulatory balance to the control of quantitative traits, hybrid vigor, genome evolution and post-zygotic speciation mechanisms. PMID- 15797618 TI - Molecular genetic approaches to developing quality protein maize. AB - Since its development more than two decades ago, Quality Protein Maize (QPM) has been adopted for cultivation in many regions of the developing world. Given the potential benefits of widespread use of QPM, research to better understand the genetic and biochemical mechanisms responsible for its altered kernel texture and protein quality is important. Recent investigations into the improved protein quality of the opaque2 mutant and the genetic mechanisms that can suppress its starchy kernel phenotype provide new insights to support the continued improvement of QPM. Chief among these developments are the use of transgenic approaches to improve nutritional quality and the discovery that an important component of modified endosperm texture in QPM is related to altered starch granule structure. PMID- 15797619 TI - Cultural transmission of fitness: genes take the fast lane. AB - Classical population genetics describes how the fate of an allele is driven by four forces: mutation, migration, selection and drift. However, these are sometimes insufficient to explain how the observed allele frequency changes and, therefore, another factor must be invoked: cultural transmission of fitness (CTF). CTF is the non-genetic transmission of any kind of behaviour that affects reproductive success. There are several clearly documented examples of CTF, and theoretical studies have shown that it affects effective population size, linkage disequilibrium and coalescent times. It is therefore a factor that must be taken into account to explain the structure of genetic diversity. In this article, we will present documented cases of how CTF affects the genetic diversity of populations and yields dramatic changes in allele frequencies. PMID- 15797620 TI - SL trans-splicing: easy come or easy go? AB - Is spliced-leader (SL) trans-splicing an ancestral eukaryotic characteristic that has been lost in multiple lineages, or did it arise independently in the various phyla in which it occurs? Recent studies have discovered SL trans-splicing in new metazoan phyla, including the chordates. Its discovery in chordates identifies, for the first time, a phylum that clearly contains both trans-splicing and non trans-splicing major groups, and defines a limited and well-understood field in which to study the evolutionary dynamics of SL trans-splicing. In this article, I summarize the evolutionarily relevant aspects of SL trans-splicing and consider the interplay among SL trans-splicing, pre-mRNA splice-signal syntax and evolutionary genomics. PMID- 15797621 TI - Genomic determinants of prognosis in colorectal cancer. AB - Colorectal cancer progression is characterized by the sequential acquisition of multiple genetic aberrations. Insights into the biology of cancer cell and the development of novel methodologies have open a new frontier in the search of independent molecular factors to better predict outcome. Besides the generation of a large list of candidate markers, their applicability in routine clinical settings has been hindered by the heterogeneity of the disease. The analysis of cumulated genetic damage offers a more comprehensive measure of the cancer cell's genomic disruption and appears as a gauge of malignant potential. The prognostic application of different determinants of genomic damage is reviewed. PMID- 15797622 TI - Inhibitory effect of celery seeds extract on chemically induced hepatocarcinogenesis: modulation of cell proliferation, metabolism and altered hepatic foci development. AB - The chemopreventive activity of methanolic extract of Apium graveolens seeds (celery seeds) has been investigated against Solt Farber protocol of hepatocarcinogenesis, oxidative stress and induction of positive foci of gamma-GT in the liver of Wistar rats. The prophylactic treatment of celery seeds extract protected dose dependently against diethylnitrosoamine (DEN)+2 acetylaminofluorine (AAF)+partial hepatectomy (PH) induced hepatocarcinogenesis and other related events such as induction of gamma-GT positive foci (P<0.001). 2 AAF administration in diet with PH in rats resulted in increased hepatic ornithine decarboxylase (ODC) activity and a consequent increase in the rate of DNA synthesis when compared to saline treated control group while pretreatment of rats with celery seeds extract resulted in inhibition of aforementioned parameters dose dependently. The augmentation of quinone reductase (QR), glutathione-S-transferase (GST) and serum gamma-glutamyl transpeptidase (GGT) activities; and depletion of the tissue GSH content after 2-AAF (i.p. injection) for five consecutive days was prevented with the administration of celery seed extract. On the basis of the above results it can be said that A. graveolens is a potent plant against experimentally induced hepatocarcinogenesis in Wistar rats. PMID- 15797623 TI - Effects of 3-methylcholanthrene on the transcriptional activity and mRNA accumulation of the oncogene hWAPL. AB - hWAPL is a human oncogene associated with uterine cervical cancer. Here, we demonstrate that hWAPL transcription is induced by 3-methylcholanthrene (3-MC) in the cervical carcinoma-derived cell line SiHa. hWAPL transcription was analyzed with evaluation of the mRNA and heterogeneous nuclear RNA (hnRNA) levels by quantitative real time PCR analysis. Flow cytometric analysis suggested that the alteration of hWAPL mRNA levels is independent of cell cycle profile. We also found that DMSO and some components of FBS affect hWAPL transcription. Interestingly, when the aryl hydrocarbon receptor (AhR) function was inhibited by alpha-naphthoflavone (ANF), the induction of hWAPL transcription by 3-MC was greater than that in AhR-functioning normal cells. These observations suggest that there are complex mechanisms regulating the transcription of hWAPL. Furthermore, mRNA level of a mouse homolog of hWAPL in mouse uterus was induced by 3-MC injection into the abdominal cavity. Thus, some effects from 3-MC exposure on uterus may be mediated by the unscheduled overexpression of hWAPL. PMID- 15797624 TI - Dietary garcinol inhibits 4-nitroquinoline 1-oxide-induced tongue carcinogenesis in rats. AB - The effects of dietary feeding with a polyisoprenylated benzophenone, garcinol, isolated from Garcinia indica fruit rind on the development of 4-nitroquinoline 1 oxide (4-NQO)-induced oral carcinogenesis were investigated in male F344 rats. At 7 weeks of age, animals were given 4-NQO at 20 ppm in the drinking water for 8 weeks to induce tongue neoplasms. They also received the diets containing 100 or 500 ppm garcinol either during (for 10 weeks) or after (for 22 weeks) the carcinogen exposure. The other rats were given tap water without 4-NQO throughout the experiment, and fed garcinol (500 ppm)-containing diet or basal diet alone. At the end of the study (week 32), incidences of tongue neoplasms and preneoplastic lesions, cell proliferation activity in the normal-like tongue epithelium estimated by 5-bromodeoxyurideine (BrdU)-labeling index and cyclin D1 positive cell ratio, and immunohistochemical expression of cyclooxygenase-2 (COX 2) in the tongue lesions were determined. Dietary garcinol significantly decreased the incidence and multiplicity of 4-NQO-induced tongue neoplasms and/or preneoplasms as compared to the control diet. Dietary administration of garcinol also significantly reduced the BrdU-labeling index and cyclin D1-positive cell ratio, suggesting reduction in cell proliferation activity in the tongue by garcinol. The COX-2 expression in the tongue lesions was also suppressed by feeding with garcinol. These results indicate that dietary administration of garcinol inhibited 4-NQO-induced tongue carcinogenesis through suppression of increased cell proliferation activity in the target tissues and/or COX-2 expression in the tongue lesions. PMID- 15797625 TI - Combined therapy with flk1-based DNA vaccine and interleukin-12 results in enhanced antiangiogenic and antitumor effects. AB - In this study, we investigated the anti-vasculature effects and the antitumor effects of combining attenuated Salmonella typhimurium vaccine strain encoding murine vascular endothelial growth factor (VEGF) receptor-2 (flk1) with plasmid DNA vector encoding the murine interleukin-12 (mIL-12) gene. In combination, flk1 based DNA vaccine and mIL-12 slowed down tumor growth more effectively than either one alone. Splenocytes from the combined group were showed a strong CTL response against both the flk1 and tumor cells. Automated image analysis revealed that the mean microvessel density was significantly reduced after administering either flk1 based DNA vaccine or mIL-12. In addition, the combination of flk1 based DNA vaccine and mIL-12 appeared more effective at reducing the microvessel density of tumor (P<0.01, both comparisons). In summary, the antivasculature effect and the anti-tumor effect were better when the combination of flk1 based DNA vaccine and IL-12 was administered in comparison to the individual treatment groups, suggesting the synergistic action of flk1 based DNA vaccine and mIL-12. PMID- 15797626 TI - Effects of progesterone on ovarian tumorigenesis in xenografted mice. AB - Circumstantial evidence indicates that progestins reduce the risk of epithelial ovarian cancer. We report that the tumorigenic capacity of human ovarian carcinoma (SKOV-3) cells inoculated into the peritoneal cavity of athymic mice is suppressed by pretreatment with subcutaneous progesterone-releasing pellets. Numbers of tumor implants on the intestines/mesentery and invasiveness into underlying host tissues were reduced at 6 weeks following exposure to progesterone. Progesterone prevented tumors from forming on the liver. Life spans of progesterone-treated animals were prolonged. There was no beneficial effect of administration of progesterone if initiated after ovarian tumors had become established on organ surfaces. Our findings implicate a role for progesterone in ovarian cancer prophylaxis. PMID- 15797627 TI - Human glutathion S-transferase A1 polymorphism and susceptibility to urothelial cancer in the Japanese population. AB - GSTA1 has been reported to be most efficient in detoxifying N-acetoxy PhIP. In this study, 341 Japanese urothelial cancer patients and 457 healthy controls were compared for frequencies of GSTA1 genotype. We present the first evidence of an association between GSTA1*B (-567G, -69T, -52A) and urothelial cancer among never smokers. The frequency of GSTA1 *A/*B or *B/*B genotype was 24.3% in urothelial cancer cases, compared with 21.2% in the control groups (OR=1.22; 95%CI 0.87 1.72) after adjustment for age, gender and smoking status. But among never smokers, the GSTA1 *A/*B or *B/*B genotype was significantly higher in urothelial cancer cases (31.2%) compared with the controls (19.9%) (OR=1.73; 95%CI 1.01 2.97). This study suggests that exposure to food-derived PhIP could be one of the risk factors in the incidence of urothelial cancer in never smokers. PMID- 15797628 TI - Breast cancer risk in a large cohort of female antidepressant medication users. AB - Paroxetine in particular has been speculated to increase the risk of breast cancer. The aim of this study was to determine if breast cancer risk is elevated among women who used paroxetine relative to those who used other antidepressants. We conducted a retrospective cohort study of 109,004 female health plan members who used various antidepressants between 1995 and 2000 to evaluate breast cancer risk. The age-adjusted relative risk comparing ever users of paroxetine to those who used other antidepressants was 1.12 (95% CI 0.96-1.31). Women who used paroxetine 2 or more years did not have a greater risk of breast cancer compared to women who used the medication for a shorter period. PMID- 15797630 TI - Apoptotic effects of extract from Antrodia camphorata fruiting bodies in human hepatocellular carcinoma cell lines. AB - The fruiting body of Antrodia camphorata is well known in Taiwan as a traditional medicine for treating cancer and inflammation. The purpose of this study was to evaluate the apoptotic effects of ethylacetate extract from A. camphorata (EAC) fruiting bodies in two human liver cancer cell lines, Hep G2 and PLC/PRF/5. Treatment with EAC decreased the cell growth of Hep G2 and PLC/PRF/5 cells in a dose dependent manner. In Fas/APO-1 positive-Hep G2 cells, EAC increased the expression level of Fas/APO-1 and its two forms of ligands, membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), in a p53-indenpendent manner. In addition, EAC also initiated mitochondrial apoptotic pathway through regulation of Bcl-2 family proteins expression, release of cytochrome c, and activation of caspase-9 both in Hep G2 and PLC/PRF/5 cells. Furthermore, EAC also inhibited the cell survival signaling by enhancing the amount of IkappaBalpha in cytoplasm and reducing the level and activity of NF-kappaB in the nucleus, and subsequently attenuated the expression of Bcl-X(L) in Hep G2 and PLC/PRF/5 cells. EAC therefore decreased the cell growth and induced apoptosis both in Hep G2 and PLC/PRF/5 cells. PMID- 15797629 TI - Frequent overexpression of cyclin D2/cyclin-dependent kinase 4 in Wilms' tumor. AB - The expression status of the three cyclin D genes (CCND1, CCND2 and CCND3), the two cyclin D-dependent kinase genes (CDK4 and CDK6) and the p16(INK4a) gene was studied in a series of 47 Wilms' tumors, 16 normal mature kidneys and two fetal kidneys. We showed predominant overexpression of CCND2 and CDK4 compared to CCND1/D3 and CDK6 respectively. We found a specific correlation between relapse and CDK4 overexpression, but not CDK6 overexpression. We did not identify any methylation of the p16(INK4a) promoter. This suggests that dysregulation of CCND2 and CDK4 plays a specific role in WT tumorigenesis. PMID- 15797631 TI - Micronuclei in EM9 cells expressing polymorphic forms of human XRCC1. AB - X-ray repair cross-complementing gene 1 (XRCC1) is involved in base excision repair (BER) through interaction with other BER enzymes, and polymorphisms in XRCC1 appear to increase the risk of various cancers. We evaluated how three XRCC1 polymorphisms, Arg194Trp, Arg280His and Arg399Gln, affect the extent of DNA damage and repair using the micronucleus assay. XRCC1 cDNAs containing the wild type sequence and the three polymorphisms were overexpressed in EM9 cells, which lack the full sequence needed to perform XRCC1 functions. Normal human XRCC1 cDNA corrected the defect in EM9 cells. Only XRCC1 cDNA containing the Arg399Gln polymorphism did not fully correct the DNA repair defect in EM9 cells. These results indicate that the Arg399Gln polymorphism, but not the Arg194Trp or Arg280His polymorphism, influences the ability of XRCC1 to repair DNA. This study may provide a model that can be used to evaluate the functional significance of polymorphisms in DNA repair genes. PMID- 15797632 TI - Overexpression of a novel superoxide-producing enzyme, NADPH oxidase 1, in adenoma and well differentiated adenocarcinoma of the human colon. AB - A superoxide-producing enzyme, NADPH oxidase 1 (Nox1), dominantly expressed in the colon, is implicated in the pathogenesis of colon cancer. Immunohistochemistry showed that Nox1 was constitutively expressed in surface mucous cells. Adenomas and well differentiated adenocarcinomas up-regulated Nox1 expression. Ki-67-negative, well differentiated tumor cells contained abundant Nox1, whereas Ki-67-positive, proliferating cells did not express it. This differentiation-dependent expression in normal as well as tumor tissues suggests distinct roles of Nox1 besides mitogenic function. Nuclear factor (NF)-kappaB was predominantly activated in adenoma and adenocarcinoma cells expressing abundant Nox1, suggesting that Nox1 may stimulate NF-kappaB-dependent antiapoptotic pathways in colon tumors. PMID- 15797633 TI - VEGF, FGF2, TGFB1 and TGFBR1 mRNA expression levels correlate with the malignant transformation of the uterine cervix. AB - Angiogenesis is a complex procedure induced by the secretion of numerous growth factors from endothelial cells. Vascular endothelial growth factor (VEGF), basic fibroblastic growth factor (FGF2), transforming growth factor-beta1, 2, 3 (TGFB1, 2, 3), and transforming growth factor-beta receptors (TGFBR1, 2, 3) mRNA expression pattern was evaluated in tissue samples with cervical intraepithelial neoplasia (CIN) and cervical cancer, compared to that of normal cervical tissues, and correlated to the clinical stage of the disease. Transcript levels of the above genes were assessed by RT-PCR analysis in a total of 44 cervical specimens. VEGF, TGFB1, TGFBR1, and FGF2 transcript levels were significantly different in the normal, CIN and cancer specimen groups (P=0.015, 0.001, 0.008, and 0.029, respectively). Higher TGFBR1 mRNA levels were observed in parallel with increased severity of the lesion, whereas FGF2 exhibited lower transcript levels. A highly significant increase of VEGF mRNA expression was found upon cervical neoplastic transformation (P<0.0001). High-grade squamous intraepithelial lesions exhibited higher VEGF mRNA levels than low-grade lesions (P=0.039). TGFBR1 and TGFBR3 receptors demonstrated significant co-expressions with TGFB2 (P<0.0001), and TGFB1 (P=0.005 and 0.002, respectively) in normal cervical specimens. However, a disruption of co-expression patterns was observed in the groups of CIN and cancer cases, compared to normal tissues. Our findings show that VEGF, FGF2, TGFB1 and TGFBR1 mRNA expression levels correlate with the malignant transformation of the uterine cervix. The involvement of the examined markers in cervical carcinogenesis is furthermore supported by the observed disruption of their mRNA co-expression patterns. PMID- 15797634 TI - Studying treatments and outcomes over time in substance abusers: Persistent Effects of Treatment Studies (PETS). PMID- 15797635 TI - Treatment outcomes: first time versus treatment-experienced clients. AB - A sample of treatment seeking clients was assessed at intake and 6, 12, 24 and 30 months later. Treatment-naive and treatment-experienced clients were compared in terms of baseline characteristics, 6-month outcomes, and predictors of these outcomes. Long-term outcomes of clients achieving an initially successful outcome were compared for the two groups. Results revealed significant short-term improvement for both groups, although treatment-experienced clients reported more serious drug use. The most consistent significant predictors of substance use outcomes were baseline severity of substance use and self-help participation. Outcome predictors unique to each group were also identified. Long-term outcomes for clients with an initial successful outcome tended to be better for treatment experienced than treatment-naive clients. Both groups tended to have poorer outcomes as the follow-up duration increased. The results suggest that common and unique predictors of outcomes be considered in treating these two groups of clients. Additionally, treatment-naive clients may be at increased risk for poor outcomes even when initial success is achieved. PMID- 15797636 TI - Transitions through homelessness and factors that predict them: three-year treatment outcomes. AB - The course of homelessness was examined among adults entering treatment in the Chicago Target Cities sample, which was aimed at improving the service delivery system in large metropolitan areas across the U.S. The objectives of the present study were: (1) Examine transitions in and out of homelessness over 3 years post entry into treatment; and (2) Determine the treatment and non-treatment factors that predict achieving and sustaining residential stability. Sixty-one percent of initially homeless participants were stably housed at 36 months. By contrast, only 14% of initially housed participants were homeless at 36 months. Sample wide, homelessness was reduced by 43% over 3 years. In conditional logistic regression models, the most consistent and persistent predictors were crack as the primary problem substance, which appears to be a risk factor for becoming and remaining homeless, and whether or not others were dependent on the participant for food/shelter, which appears to be a protective factor for achieving housing and preventing homelessness. In general, specific treatment factors did not predict outcomes. Limitations and implications for treatment are discussed. PMID- 15797637 TI - Gender differences in long-term drug treatment outcomes in Chicago PETS. AB - Few long-term follow-up studies of substance abusers have examined gender differences. In the current study, gender differences were examined at 36 months following residential or outpatient drug-free treatment among 951 participants in the Chicago Target Cities Project, the majority of whom were female (62%) and African American (93%). There were no differences in the proportion of men and women who reported any alcohol or drug use at the 36-month follow-up, with an overall reduction of 41% from intake. Greater proportions of men were incarcerated or employed, whereas greater proportions of women had returned to treatment, lived with their children, lived with a substance user, or had interpersonal problems. Women, as a group, had greater increases over time in self-help participation, free time spent with family, non-using family/friends, and employment. Although both men and women showed significant improvements following treatment, gender differences persisted in several areas of psychosocial functioning related to recovery. PMID- 15797638 TI - Economic benefits of substance abuse treatment: findings from Cuyahoga County, Ohio. AB - We estimated long-term economic benefits and treatment costs for a sample of substance abuse clients who received treatment in Cuyahoga County, Ohio, using health, criminal activity, and earnings data from the Persistent Effects of Treatment Studies. Clients were interviewed at baseline and 6, 12, 24, and 30 months following baseline. We find positive benefits from substance abuse treatment, almost of all of which were derived from reduced criminal activity and increased real earnings, with overall benefit-to-cost ratios ranging from 2.8 to 4.1. The reductions in costs to society were found to be persistent over the long term, 30-month follow-up period. On average, treatment was found to be cost beneficial regardless of the number of times a client entered treatment in the baseline or follow-up periods. Clients who entered residential treatment and then step down to less intensive care showed greater treatment benefits than clients who only received residential treatment. PMID- 15797639 TI - The duration and correlates of addiction and treatment careers. AB - While addiction is increasingly recognized as a chronic condition, little information is available on the actual duration of addiction and treatment careers. The purpose of this study was to estimate the duration and correlates of years between (a) first use and at least a year of abstinence and (b) first treatment admission and at least one year of abstinence. Data are from 1,271 of 1,326 (96%) people recruited from a stratified sequential sample of admissions to publicly funded treatment programs in a large metropolitan area. Participants were interviewed at 6, 18, 24, 36, 48, and 60 months post-intake. With an average age at the referent intake of 35 (SD = 8) years, the sample is 59% female, 87% African American, 7% Hispanic, and 5% white. The most common dependence diagnoses were for cocaine (64%), alcohol (44%), opioids (41%), and/or marijuana (14%). Using lifetime substance use and treatment histories collected at intake and subsequent treatment utilization recorded during follow-up interviews, we conducted survival analysis to estimate the time from first use and first treatment until people reported 12 months of abstinence or died (with people still using, in treatment, or dead at the last follow-up treated as right censored). During the three years after intake, 47% reached at least 12 months of abstinence. The median time from first to last use was 27 years. The median time from first treatment episode to last use was 9 years. Years to recovery were significantly longer for males, people starting use under the age of 21 (particularly those starting under the age of 15), people who had participated in treatment 3 or more times, and for people high in mental distress. The exploratory results suggest that multiple episodes of care over several years are the norm and that rather than thinking of multiple episodes in terms of "cumulative dosage," it might be better thought of as further evidence of chronicity and that we need to develop and evaluate models of longer term recovery management. PMID- 15797640 TI - Pathways in the relapse--treatment--recovery cycle over 3 years. AB - For many individuals, substance use leads to a chronic cycle of relapse, treatment reentry, and recovery, often lasting for decades. This study replicates earlier work, documents the transition patterns within the cycle during a 3-year period, and identifies variables that predict these transitions. Data are from 1,326 adults recruited from sequential admissions to 12 substance abuse treatment facilities in Chicago, IL, between 1996 and 1998. Participants were predominantly female (60%) and African American (88%) adults. Participants were interviewed at intake, and at 6, 24, and 36 months post-intake follow-up rates ranged from 94% to 98% per wave. At each observation, participants' current status in the cycle was classified as (1) in the community using, (2) incarcerated, (3) in treatment, or (4) in the community not using. The transitional probabilities and correlates of pathways between these states were estimated. Over 83% of the participants transitioned from one point in the cycle to another during the 3 years (including 36% two times, 14% three times). For the people in the community, about half remained in the same status (either using or abstinent) and just under half transitioned. The majority of people whose beginning status was incarceration or in-treatment also transitioned by the end of the observation period. While there was some overlap, predictors typically varied by pathway and direction (e.g., using to not using vs. not using to using). These results help demonstrate the need to adopt a chronic vs. acute care model for substance use. While exploratory and observational, several of the predictors are time-dependent and identify promising targets for interventions designed to shorten the cycle and increase the long-term effectiveness of treatment. PMID- 15797642 TI - Contemporaneous relationship between substance abuse treatment and poly-substance use: evidence from the Persistent Effect of Treatment Studies. AB - Data from the Persistent Effect of Treatment Studies (PETS) are used to explore the relationship between duration of substance use treatment and simultaneous poly-substance using behaviors. Studying such contemporaneous relationships is especially important given the chronic nature of the substance-using population (McLellan, 2002) in the PETS study. Data were collected at intake to treatment programs and follow-up interviews were performed periodically at treatment program sites. One of the features of the analysis was the development of a poly substance scale to measure multiple substance use. Multilevel models were implemented to examine the relationship between three levels of care (i.e., intensive outpatient, outpatient, and residential) and simultaneous poly substance use. Contemporaneous effects of treatment were obtained such that higher duration of treatment was associated with drops in substance-using behaviors. This result supports the need for sustaining treatment for a population of chronic substance abusers. PMID- 15797641 TI - Predictors of alcohol and crack cocaine use outcomes over a 3-year follow-up in treatment seekers. AB - This study identified predictors of long-term alcohol and crack cocaine use outcomes in individuals participating in the Persistent Effects of Treatment Study. The domains that were assessed included motivation, self-efficacy, social support, psychiatric severity, employment, housing status, and self-help group attendance at baseline and 6, 12, 24, and 30 month follow-ups. In alcohol users, higher perceived seriousness of substance use problems, self-efficacy, and self help group attendance, as well as lower social support for substance use, consistently predicted better alcohol use outcomes in the subsequent assessment period. In crack cocaine users, only self-efficacy consistently predicted cocaine use outcomes. Higher self-efficacy during follow-up was predicted by lower perceived seriousness of substance use and lower alcohol use frequency in the prior assessment period, whereas greater self-help group attendance was predicted by greater perceived seriousness of substance use, and lower substance use frequency. PMID- 15797643 TI - Ethical issues in the conduct of longitudinal studies of addiction treatment. AB - Many complex ethical issues arise in the day-to-day conduct of longitudinal studies of addiction treatment. These issues are rooted, in part, in the sustained and potentially ambiguous relationship between research staff and study participants, the frequently changing clinical and legal status of study participants, the assertive methods required to generate high follow-up rates, and the numerous systems of care and control in which participants are involved. To identify common ethical issues that arise in such studies, the authors conducted individual and group interviews with seasoned members (case trackers, field trackers, interviewers, and supervisors) of the research team. The ethical dilemmas identified through these interviews fell into seven broad arenas: (1) informed consent for research participation, (2) confidentiality and information disclosure, (3) relationship boundaries between study participants and research staff, (4) duty to warn/report responsibilities, (5) questions of autonomy and privacy, (6) issues related to compensation for research participation, and (7) data integrity. Case studies that illustrate common ethical dilemmas within each of these seven areas are presented and discussed. Ethical dilemmas in the study of addiction treatment can be effectively managed via ethically informed research protocols, staff training in ethical decision-making, monitoring and supervision, and collective debriefing of critical events. PMID- 15797644 TI - Retinal arteriolar narrowing and hypertension. PMID- 15797645 TI - Angiotensin II increases the expression of lectin-like oxidized low-density lipoprotein receptor-1 in human vascular smooth muscle cells via a lipoxygenase dependent pathway. AB - BACKGROUND: Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a membrane protein that can act as a surface endocytosis receptor for oxidized LDL (ox-LDL). As increased cellular uptake of ox-LDL by macrophages and activated smooth muscle cells may transform these cells into foam cells, potential interactions among LDL oxidation, ox-LDL uptake, and regulators of vascular smooth muscle cell function are of obvious interest. The objective of this study was to examine the effect of angiotensin II (AII) on the expression of LOX-1 and ox-LDL degradation in human vascular smooth muscle cells (VSMC) METHODS: We performed in vitro experiments in a human VSMC line (T/G HA-VSMC) derived from normal aortic VSMC, using standards methods. RESULTS: We found that AII (10(-7) mol/L) increased the expression of LOX-1 (approximately 2.5-fold, P < .0001) in association with higher degradation of ox-LDL by HA-SMC (from 4019 +/- 529 ng/mg cell protein to 6207 +/- 287 ng/mg cell protein; P = .0033). AII also increased the expression of 12-lipoxygenase (12-LO) and 15-lipoxygenase (15-LO) by approximately 2.2-fold (P = .03) and approximately 3-fold (P = .006), respectively. In addition, AII (10(-7) mol/L) increased the release of 12- and 15 hydroxyeicosatetraenoic acid from VSMC within 10 min approximately 3-fold (P = .03) and 50% (P < .05), respectively. CONCLUSIONS: Our study findings provide evidence that angiotensin II upregulates LOX-1 and 12-LO and 15-LO expression in human VSMC, thereby potentially providing mechanisms for both accelerated LDL oxidation within the cell and the internalization of exogenous ox-LDL, two processes that could increase the susceptibility of human VSMC to further transformation into foam cells. PMID- 15797647 TI - Vascular abnormalities in asymptomatic, healthy young adult smokers without other major cardiovascular risk factors: the Bogalusa Heart Study. AB - Cigarette smoking, an established cardiovascular (CV) disease risk factor, is known to impair pulsatile arterial function in middle-aged and older adults. However, information is scant in healthy young adults for whom smoking is the only CV risk factor, at current guidelines. Nonsmokers (n = 145) and smokers (n = 142) aged on average 36 years were selected for not having obesity, hypertension, dyslipidemia, diabetes, or clinically manifest CV disease. Pulsatile arterial function was measured in terms of large artery compliance (C1), small artery compliance (C2), and systemic vascular resistance (SVR) by noninvasively recorded radial artery waveforms. Smokers versus nonsmokers had significantly lower measures of adiposity and LDL-cholesterol; and higher systolic blood pressure and triglycerides. In addition, smokers versus nonsmokers had lower C2 (5.09 v 6.63 mL/mm Hg x 100, P = .0009) and higher SVR (1399.0 v 1325.5 dyn . sec . cm(-5), P = .006), after adjustment for race, sex, and age. Decreases in C2 (P for trend = .001) and increases in SVR (P for trend = .01) were noted with increasing years of smoking. Multivariate analysis revealed that duration of smoking was associated adversely with C2 (P = .004), independent of race, sex, age, systolic and diastolic blood pressures, HDL-cholesterol, triglycerides, glucose, and insulin. The odds of having adverse C2 (bottom 10 percentile) and SVR (top 10 percentile) were, respectively, 2.9 (P = .01) and 2.6 (P = .07) times higher in smokers versus nonsmokers. The observed deleterious effects of cigarette smoking on arterial wall dynamics in otherwise healthy young adults underscore the need for aggressive early prevention and intervention strategies to control smoking behavior. PMID- 15797646 TI - Regression of carotid atherosclerosis by control of morning blood pressure peak in newly diagnosed hypertensive patients. AB - BACKGROUND: Morning blood pressure (BP) peak may be a risk factor for cardiovascular disease. Whether morning BP should be a target of hypertension treatment is not known. We investigated the relationship between morning BP variations, carotid internal-medial thickness (CIMT), circulating inflammatory markers, and sympathetic activity in hypertensive patients with different patterns of morning BP increase at baseline and after antihypertensive treatment. METHODS: One hundred twenty-eight hypertensive patients with morning BP peak (MP+) were compared with 196 hypertensive patients without morning BP peak (MP-). All patients performed 24-h ambulatory BP monitoring, assessment of CIMT, circulating concentration of C-reactive protein (CRP), interleukin-6 (IL-6), interleukin-18 (IL-18), and urinary catecholamines. RESULTS: Compared with MP- patients, MP+ patients had higher CIMT and urinary catecholamine output (P < .001), as well as CRP, IL-6, and IL-18 (P < .001). We randomly assigned 128 drug naive MP+ patients to either metoprolol or carvedilol, two antihypertensive drugs with different effects on sympathetic activity. The primary outcome was change in CIMT and circulating inflammatory markers at 12 months. Morning BP decreased more among patients in the carvedilol group (P < .001), whereas clinic BP showed a similar decrease in both groups. The CIMT (P < .001), IL-6 (P < .001), IL-18 (P < .001), and CRP (P < .001) decreased more in the carvedilol group than in the metoprolol group. The CIMT regression was observed in 49% of patients in the carvedilol group and 18% of patients in the metoprolol group (P < .01). Reduction in CIMT was directly associated with changes in morning BP. CONCLUSIONS: Higher CIMT and circulating inflammatory markers coexist in hypertensive patients with morning BP peak, and might contribute to their increased cardiovascular risk. Carotid atherosclerosis can be prevented by control of morning BP. PMID- 15797648 TI - Circulating matrix metalloproteinase-9 and tissue inhibitors of metalloproteinases-1 and -2 levels in gestational hypertension. AB - BACKGROUND: Gestational hypertension (GH) is dangerous to both mother and child. Arterial invasiveness and growth are dependent on successful extracellular matrix (ECM) breakdown, which may be abnormal in GH. We hypothesized abnormalities in circulating matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinases-1 and -2 (TIMP-1 and TIMP-2, respectively) in patients with GH, when compared with normotensive women with normal pregnancies and healthy nonpregnant control subjects. METHODS: Plasma MMP-9, TIMP-1, and TIMP-2 were measured by ELISA in 23 women with GH, 30 normotensive pregnant women, and 28 nonpregnant women who were matched for age, gestational age, and parity. RESULTS: Levels of circulating MMP-9, TIMP-1 and TIMP-2, and the MMP-9/TIMP-1 and MMP 9/TIMP-2 ratios were significantly different among the three groups (P = .026, P = .006, P = .007, P = .001 and P = .008 respectively). Within the GH group, MMP-9 and the MMP-9/TIMP-1 ratio correlated negatively with age (r = -0.581, P = .004 and r = -0.563, P = .005, respectively) and levels of diastolic blood pressure (r = -0.432, P = .040 and r = -0.461, P = .027, respectively). With multiple regression analysis, only age independently correlated with circulating levels of MMP-9 (P = .010); neither age nor levels of diastolic blood pressure had any effect on the MMP-9/TIMP-1 ratio. CONCLUSIONS: We have demonstrated altered MMP/TIMP ratios in maternal blood during GH. These observations suggest pregnancy related changes in ECM breakdown and turnover. Given the importance of changes in ECM composition to vascular and cardiac structure in hypertension, we suggest that these observations may be related to the pathophysiology of human GH. PMID- 15797649 TI - Maternal autoantibodies from preeclamptic patients activate angiotensin receptors on human mesangial cells and induce interleukin-6 and plasminogen activator inhibitor-1 secretion. AB - BACKGROUND: Preeclampsia affects 3-5% of all pregnancies. It is a major cause of maternal and fetal morbidity and mortality. Recent studies demonstrate that autoantibodies against the angiotensin II type 1 (AT(1)) receptor are present in the serum of preeclamptic patients. In this study, we investigated the role of AT(1) receptor-agonistic autoantibody (AT1-AA) regarding interleukin-6 (IL-6) and plasminogen activator inhibitor-1 (Pai-1) secretion in human mesangial cells. METHODS: The study included ten patients: five severely preeclamptic and five normotensive pregnant women. Immunoglobulin-G (IgG) was purified from each individual. The presence of AT1-AA was determined based on its ability to stimulate an increase in the contraction rate of rat neonatal cardiomyocytes. Primary human mesangial cells were chosen to study IgG-induced secretion of IL-6 and Pai-1. Losartan and epitope peptides were used to determine whether AT1-AA interaction with AT(1) receptor was associated with stimulation of IL-6 and Pai-1 secretion and was mediated through AT(1) receptor activation. RESULTS: The IgG from preeclamptic patients stimulated an increased contraction rate in rat neonatal cardiomyocytes. The IgG from preeclamptic patients induced the AT(1) receptor-specific secretion of IL-6 and Pai-1 from human mesangial cells at a significantly higher level than that achieved with IgG from normotensive patients. Competition with an epitope peptide suggested that the AT(1) receptor was stimulated by AT1-AA. CONCLUSIONS: Our findings suggest that a maternal autoantibody with the ability to activate AT(1) receptors may account for the development of renal damage seen in preeclamptic patients. PMID- 15797650 TI - Arterial stiffness and fetal growth in normotensive pregnancy. AB - BACKGROUND: Normal pregnancy is characterized by a decrease in peripheral resistance and generalized vasodilation resulting in plasma volume expansion, which is associated with intrauterine growth. Stiffness of the arterial system may be a measure of the degree of plasma volume expansion. Pulse wave velocity (PWV), measured by applanation tonometry, is a validated approach to determine arterial stiffness. Pulse pressure (PP) is considered a surrogate measure for arterial stiffness. The aim of this study was to evaluate the association between arterial stiffness and fetal growth. METHODS: In 50 normotensive pregnancies, carotid-femoral PWV was measured in the third trimester in 30 degrees lateral position. Blood pressure measurements were performed with conventional auscultatory sphygmomanometry. Birth weight centiles and weight centiles at the age of 6 months were recorded. Linear regression models were used for statistical analyses. RESULTS: There was a significant relationship in PWV with both birth weight centiles and catch-up growth after birth, independent of mean arterial pressure (MAP). An increase of 1 m/sec in PWV was associated with a decrease in birth weight centiles by 17.6% and a catch-up of 22.3% in weight centiles after birth. A stronger association was found for pulse pressure and birth weight centiles. An increase of 1 mm Hg was associated with a decrease in birth weight centiles by 1.8%. There was no association between MAP and birth weight centiles. CONCLUSIONS: In normotensive pregnancy arterial stiffness is associated with birth weight centile and catch-up growth after birth, independently from MAP. This suggests that arterial stiffness reflects maternal vascular adaptation to pregnancy better than blood pressure. PMID- 15797651 TI - Mild renal dysfunction is associated with electrocardiographic left ventricular hypertrophy. AB - BACKGROUND: Both renal dysfunction and left ventricular hypertrophy (LVH) are signs of end-organ damage, risk markers of cardiovascular (CV) disease and chronic heart failure. In selected populations such as those with diabetes or hypertension, renal dysfunction was found to be related to LVH. We studied the relation between renal dysfunction and LVH in a cross-sectional study in 8592 inhabitants from Groningen, The Netherlands. METHODS: Standard 12-lead electrocardiograms were recorded, and LVH was classified using the Cornell voltage duration product. Renal dysfunction was defined as creatinine clearance <60 mL/min/1.73 m(2) or microalbuminuria (30 to 300 mg/24 h). RESULTS: Electrocardiographic signs of LVH were present in 396 of subjects (5.3%). Subjects with LVH were older and had a more extensive CV risk profile. We found that LVH was more prevalent in subjects with renal dysfunction than in those without (8% v 4%, P < .001). Multivariate regression analysis demonstrated that renal dysfunction was independently related to a 1.47-fold increased risk of the presence of LVH (95% CI = 1.15 to 1.88, P = .009). In addition, both creatinine clearance (OR = 1.56, 95% CI = 1.07 to 2.2, P = .044) and microalbuminuria (OR = 1.37, 95% CI = 1.04 to 1.80, P = .024) were independently associated with the presence of LVH. CONCLUSION: Subjects with mild renal dysfunction have a substantially higher risk of LVH on electrocardiography than those without renal dysfunction. PMID- 15797653 TI - Atrial and brain natriuretic peptides as markers of cardiac load and volume retention in primary aldosteronism. AB - BACKGROUND: Atrial and brain natriuretic peptides (ANP and BNP, respectively) are cardiac hormones with natriuretic and vasodilator actions. The present study was carried out to determine the natriuretic peptide that is closely related to cardiac load and volume retention in patients with primary aldosteronism (PA). METHODS: We examined 11 patients with PA due to aldosterone-producing adrenal adenoma before and after surgical resection. Plasma levels of ANP and BNP were measured by specific immunoradiometric assays, and total blood volume was determined by a plasma tracer method with (131)I-human albumin. RESULTS: Plasma levels of ANP and BNP were elevated in the PA patients compared with normotensive control subjects (P < .01), and the elevated ANP and BNP levels were reduced (P < .01) after adenoma resection. When analyzed with the pre- and postoperative data, a significant relationship was observed between mean blood pressure and plasma levels of ANP (r = 0.64, P < .01) and BNP (r = 0.58, P < .01). The BNP was significantly correlated with the SV1 + RV5 voltage (r = 0.65, P < .01) and with total blood volume (r = 0.57, P < .01), but this was not the case for ANP. CONCLUSION: The present results suggest that the plasma level of BNP is more closely related to cardiac load or volume status than ANP is, in patients with PA due to adrenal adenoma. PMID- 15797652 TI - Insulin-induced changes in beta-adrenergic response: an experimental study in the isolated rat papillary muscle. AB - BACKGROUND: The aim was to investigate the ability of insulin to modulate the response to beta-adrenergic action on myocardial contractility, assessed as percentage changes of developed tension, in isolated rat papillary muscle. METHODS: Dose-response curves for isoproterenol, calcium, and forskolin were constructed in an incremental fashion with the presence or absence of insulin at the dose of 50 muU/mL. Dose-response curves for isoproterenol on insulin background were also assessed in the presence and absence of a selective antagonist for beta(2)-adrenoceptor, ICI, at the dose of 5 x 10(-8) mol/L. RESULTS: Insulin did not modify the dose-response curve to calcium (EC(50): 1.4 +/- 0.4 mmol/Lfor insulin, n = 8 v 1.5 +/- 0.3 mmol/L for control, n = 8; P = not significant), whereas it was able to shift to the left the dose-response curve and reduce significantly the EC(50) of isoproterenol (EC(50): 0.2 +/- 0.2 nmol/L for insulin, n = 13 v 1.1 +/- 0.4 nmol/L for control, n = 12; P < .01). ICI shifted to the right dose-response curve of isoproterenol and increased about 10 fold the EC(50) value of isoproterenol, but insulin was still able to shift to the left dose-response curve of isoproterenol and to reduce significantly the EC(50) of isoproterenol also in the presence of ICI (EC(50): 11.0 +/- 1.5 nmol/L for ICI, n = 7 v 1.9 +/- 0.8 nmol/L for ICI + insulin, n = 7; P < .01). Insulin did not modify the dose-response curve to forskolin. CONCLUSIONS: Our results suggest that the insulin-induced modulation of contractility is calcium independent and insulin leads to a supersensitization on the beta(1) adrenoceptors without effects on beta-adrenoceptor independent adenylate cyclase related pathway. PMID- 15797654 TI - The effect of losartan and losartan/hydrochlorothiazide fixed-combination on magnesium, zinc, and nitric oxide metabolism in hypertensive patients: a prospective open-label study. AB - BACKGROUND: Thiazide diuretics and angiotensin-converting enzyme inhibitors can cause excessive urinary zinc (Zn) loss and Zn depletion. Thiazides may also induce magnesium (Mg) deficiency, which may exacerbate hypertension. Data on the effects of angiotensin receptor blockers on Zn and Mg homeostasis are scarce. METHODS: Seventeen hypertensive patients were studied (ten men and seven women, age 50 +/- 3 years, blood pressure 158 +/- 5 / 95 +/- 3 mm Hg). Patients were treated with losartan 50 mg/day for 4 weeks followed by a fixed combination of 50 mg losartan and 12.5 mg hydrochlorothiazide for 4 weeks more. Blood and 24-h urine were collected at baseline and after each study period. Zinc and Mg levels were measured in serum, urine, and peripheral blood mononuclear cells. Nitric oxide metabolites were measured in urine. RESULTS: Treatment with losartan resulted in a significant increase in the urinary Zn/creatinine ratio (from 0.020 +/- 0.004 microg/mg to 0.034 +/- 0.005 microg/mg, P = .02), which was further increased by the losartan/hydrochlorothiazide combination (from 0.034 +/- 0.005 microg/mg to 0.053 +/- 0.008 microg/mg, P = .03). Serum Zn levels were significantly decreased after losartan/hydrochlorothiazide (from 80.0 +/- 4.0 microg/dL at baseline to 74.0 +/- 3.0 microg/dL, P = .007). Peripheral blood mononuclear Zn concentrations were decreased also, but this was not statistically significant. Serum, urinary, and peripheral blood mononuclear Mg levels were not significantly affected by treatment. Nitric oxide urinary metabolites were unchanged throughout the study. CONCLUSIONS: Treatment with losartan causes an increase in urinary Zn excretion and induces Zn deficiency in patients with hypertension. The addition of hydrochlorothiazide has an additive effect. Magnesium and nitric oxide metabolism are not affected by either treatment. PMID- 15797655 TI - Calcimimetic NPS R-568 induces hypotensive effect in spontaneously hypertensive rats. AB - BACKGROUND: The discovery of calcium receptors and calcimimetics created the possibility of "pharmacologic parathyroidectomy" (phPTX), which decreased secretion of parathormone (PTH). Parathyroid glands of spontaneously hypertensive rats (SHR) and of patients with primary hyperparathyroidism and hypertension secrete parathyroid hypertensive factor (PHF). Parathyroidectomy decreases blood pressure in these rats and in patients. The present study determined whether phPTX induced by calcimimetics decreases mean arterial blood pressure (MAP) in hypertensive rats. METHODS: Hypertensive SHR and normotensive Wistar Kyoto (WKY) rats were used. Clearance experiments were performed and the effect of 1 mg/kg body weight (given intravenously) synthesized NPS R-568 (NPS) on MAP in the presence or absence of thyroparathyroidectomy (TPTX) was monitored. RESULTS: The success phPTX and TPTX were proven by a significant decrease in plasma Ca(2+) concentration and a decrease in urinary fractional phosphate excretion (FE Pi). The administration of NPS significantly decreased blood pressure in SHR versus SHR/control: Delta(0-50 min of experiment) MAP -16.5 +/- 2.5 mm Hg v -3.2 +/- 1.5 mm Hg (P < .002). The TPTX decreased blood pressure in SHR versus SHR/control and was not different versus SHR/TPTX/NPS (DeltaMAP: -10.2 +/- 1.6 mm Hg v -3.2 +/- 1.5 mm Hg (P < .01) and v -8.3 +/- 2.2 mm Hg (P = not significant). In normotensive WKY rats application of NPS did not reach significance in DeltaMAP: 6.7 +/- 1.8 mm Hg v -2.6 +/- 2.8 mm Hg (P = not significant) in WKY/control. The TPTX lowered blood pressure in WKY versus WKY/control and remained unchanged versus WKY/TPTX/NPS (DeltaMAP: -11.3 +/- 1.7 mm Hg v -2.6 +/- 2.8 mm Hg (P < .04) and v -11.4 +/- 2.6 mm Hg (P = not significant). CONCLUSIONS: We conclude that phPTX with NPS R-568 is responsible for a decrease of MAP in SHR. PMID- 15797656 TI - Risk reduction therapy for syndrome X: comparison of several treatments. AB - BACKGROUND: Syndrome X, also termed the metabolic syndrome, is a cluster of physiologic and metabolic abnormalities including abdominal obesity, hyperinsulinemia, dyslipidemia and hypertension. Severe cardiovascular morbidity is associated with this pre-diabetic syndrome. We recently suggested that hyperhomocysteinemia is affiliated with this syndrome and thereby contributes to the vascular risk accompanying this condition. The present study compared the effects of antihypertensive, lipid-lowering, and insulin-sensitizing agents on the above-mentioned components of this metabolic syndrome. METHODS: Experimental metabolic syndrome was induced in Sprague-Dawley rats by feeding them a fructose enriched diet (FED) for 5 weeks. During the last 2 weeks, the rats were treated with amlodipine, captopril, bezafibrate, or rosiglitazone in addition to FED. The control group did not receive any medication. Another control group was fed standard rat chow for 5 weeks. Post- and pretreatment measurements of body weight, systolic blood pressure (SBP), fasting plasma insulin, triglycerides, and total homocysteine concentrations were compared. RESULTS: Amlodipine reduced SBP but did not show metabolic impact. Bezafibrate improved SBP, triglycerides, and insulin but induced elevation of homocysteine levels. Captopril and rosiglitazone remarkably improved SBP, insulin, triglycerides, and total homocysteine levels. In addition, rosiglitazone alone promoted weight gain. CONCLUSIONS: The results indicate that captopril and rosiglitazone have a greater cardiovascular protective potential than amlodipine or bezafibrate. Captopril would be the best choice for patients with metabolic syndrome in whom hypertension and obesity are prominent, whereas rosiglitazone would be the preferred drug when glucose and other metabolic parameters are disturbed. PMID- 15797657 TI - Significance of elevated blood pressure and its management on the short-term outcome of patients with acute ischemic stroke. AB - BACKGROUND: The objectives of this study were to characterize blood pressure (BP) in acute ischemic stroke and to determine its relationship with short-term functional outcome. METHODS: We examined 24-h BP recordings in 434 patients with ischemic stroke (lacunar stroke [LS], n = 205; non-lacunar stroke [NLS], n = 229) and in 178 normotensive subjects. Stroke severity was evaluated by the National Institutes of Health Stroke Scale (NIHSS). Patients found to be hypertensive on BP recordings on day 1 were given captopril or amlodipine. The primary outcome was both moderate-to-severe disability (Rankin scale scores 4 to 6) on day 7 or death during hospital stay. RESULTS: Patients with LS and NLS had significantly higher systolic BP (SBP) and diastolic BP levels than control subjects. On day 1, patients with NLS showed significantly higher NIHSS scores, SBP, and heart rate (HR) levels than LS patients. In the multivariate analysis, combined death or dependency was associated with NIHSS score (odds ratio [OR] = 1.08 per 1-point increase, 95% confidence interval [CI] = 1.04 to 1.13), 24-h SBP >160 mm Hg (OR = 2.35, 95% CI = 1.10 to 5.52), and plasma glucose levels >125 mg/dL on admission (OR = 1.88, 95% CI =1.03 to 3.57), whereas a decrease in SBP on day 7 (OR = 0.46, 95% CI = 0.24 to 0.88) was associated with better short-term outcome. CONCLUSIONS: At presentation, NLS patients showed higher BP levels than LS patients. Moderate reductions in BP during the first week after admission were associated with short-term functional improvement in patients with acute ischemic stroke. PMID- 15797658 TI - Lower hypertension risk in Mexico City than in San Antonio. AB - BACKGROUND: We examined the effects of blood pressure (BP), weight, and weight gain on hypertension risk in two similar ethnic origin populations, subjects in Mexico City and Mexican Americans in San Antonio. METHODS: The Mexico City Diabetes Study and San Antonio Heart Study are population-based, epidemiologic studies with identical survey protocols. Incident hypertension (BP > or = 140/90 mm Hg or current antihypertensive treatment) was analyzed in subjects aged 35 to 64 years of Mexican ethnicity living in low-income neighborhoods (n = 1467 in Mexico City, n = 628 in San Antonio). RESULTS: In Mexico City, 10.6% of men and 13.1% of women developed hypertension in a 6.5-year period; in San Antonio, 28.6% and 28.7% in a 7.5-year period, respectively. Poisson regression analysis demonstrated a greater hypertension risk in San Antonio for both men (risk ratio [RR] = 1.75, 95% confidence interval [CI]: 1.19-2.56) and women (RR = 1.40, 95% CI: 1.05-1.86). In a multiple linear regression analysis, systolic BP change was associated with weight gain in Mexico City (P < .001 in men and women) and San Antonio (P = .045 in men, and P = .027 in women) independently of age, BP, obesity, alcohol consumption, cigarette smoking, diabetes, and antihypertensive treatment. These covariates did not fully explain greater increments of systolic BP in San Antonio than in Mexico City (P < .001 in men and women). CONCLUSIONS: Hypertension risk is lower in Mexico City than in San Antonio. Systolic BP increases with weight gain, independently of other determinants of hypertension. PMID- 15797659 TI - beta(2)-Adrenergic receptor polymorphisms and determinants of cardiovascular risk: the Cardiovascular Health Study. AB - BACKGROUND: Common Arg16Gly and Gln27Glu polymorphisms of the beta(2)-adrenergic receptor (beta(2)AR) have been associated with hypertension and coronary disease. This analysis of older adults in the Cardiovascular Health Study examined whether these polymorphisms were associated with blood pressure (BP), subclinical atherosclerosis, and, among treated hypertensive individuals, differences in coronary disease risk according to antihypertensive drug class. METHODS: Altogether, 5249 participants (4441 white and 808 African American, median follow up time 10.2 years) were genotyped for both polymorphisms. Ankle-arm index (AAI), carotid intima-media thickness (IMT), and brachial flow-mediated dilation were measured cross-sectionally. All estimates were adjusted for ethnicity. RESULTS: Relative to Gln27 homozygotes, carrying the Glu27 allele was not associated with new-onset hypertension (hazard ratio [HR] = 1.01, 95% confidence interval [CI] = 0.87 to 1.16), BP control (odds ratio [OR] = 0.97, 95% CI = 0.89 to 1.06), AAI (mean difference 0.0042 +/- 0.0052), carotid IMT (mean difference 0.0044 +/- 0.02 mm), or brachial flow-mediated dilation (mean difference in baseline diameter 0.028 +/- 0.036 mm; the most marked of three measures). Among treated hypertensive individuals, coronary disease risk was similar in Glu27 carriers relative to Gln27 homozygotes in subgroups defined by use of beta-blockers (HR = 1.09, 95% CI = 0.64 to 1.87) or other antihypertensive medications (HR = 1.00, 95% CI = 0.78 to 1.28). Results were similar for the Arg16Gly polymorphism. CONCLUSIONS: The association of beta(2)AR genotype with coronary disease previously reported in this older adult population is not likely to be explained by BP levels, subclinical atherosclerosis, or antihypertensive treatment. Other measures of vascular response, gene-gene or gene-environment interactions, or characteristics developing earlier in life may mediate the association between beta(2)AR genotype and coronary disease and merit further research. PMID- 15797660 TI - Lack of agreement between office and ambulatory blood pressure responses to hydrochlorothiazide. AB - BACKGROUND: Differences between the antihypertensive responses to drug therapy measured by office blood pressure (OBP) and ambulatory blood pressure monitoring (ABPM) techniques have been noted but rarely analyzed. We studied whether the OBP and 24-h ABPM responses to hydrochlorothiazide differ and, if so, the relevance of these differences. METHODS: The OBP and ABPM responses to hydrochlorothiazide (25 mg/d, for 4 weeks) were measured in 228 subjects with essential hypertension, and mean responses were compared between methods using the Student paired t test. To assess variation in the agreement between OBP and ABPM responses among subjects, the limits of agreement were calculated as the mean difference between OBP and ABPM responses +/-2 standard deviations. RESULTS: The mean systolic OBP response was 4.8 mm Hg greater than the response measured by ABPM (-14.3 v -9.5 mm Hg, P < .001), and the mean diastolic OBP response was 2.1 mm Hg greater than the response measured by ABPM (-7.5 v -5.5, P < .001). The limits of agreement between the OBP and ABPM responses ranged from -18.7 to +28.2 mm Hg for systolic response and from -12.9 to +17.1 mm Hg for diastolic response. The systolic and diastolic OBP and ABPM responses were in opposite directions in 22.8% and 23.7% of the subjects, respectively. CONCLUSIONS: Compared to ABPM, OBP overestimates the mean systolic and mean diastolic blood pressure responses to hydrochlorothiazide. Variation among subjects in the magnitude and direction of responses renders OBP an unreliable predictor of ABPM responses. PMID- 15797661 TI - Association study of calcitonin-receptor-like receptor gene in essential hypertension. AB - BACKGROUND: Plasma adrenomedullin (ADM) concentrations increase in patients with hypertension, renal failure, heart failure, essential pulmonary hypertension, myocardial infarction, endotoxin shock, and many other conditions. The ADM receptor is a complex molecule that consists of calcitonin-receptor-like receptor (CRLR) and receptor activity-modifying protein 2 (RAMP2). Because CRLR determines the binding specificity of ADM, the CRLR gene is thought to be a susceptibility gene of hypertension. However, studies have not yet defined the relationship between the CRLR gene and hypertension. The aim of the present study was to investigate relationships between single-nucleotide polymorphisms (SNP) in the human CRLR gene and essential hypertension (EH) in a Japanese population. METHODS: We selected four SNP in the human CRLR gene (rs3771073, rs696574, rs698590, and rs1528233), and we performed a genetic association study in 209 EH patients and 216 age-matched normotensive (NT) individuals. RESULTS: There was no significant difference in overall distribution of genotypes or alleles of any of the SNP between the EH and NT groups. However, among women, the T allele of the SNP rs696574 (C --> T, in intron 6) was significantly more frequent in EH subjects than in NT subjects (P = .032). CONCLUSION: Our findings suggest that rs696574 can be used as a genetic marker of EH in women. PMID- 15797662 TI - Dyslipidemic hypertension: distinctive features and cardiovascular risk in a prospective population-based study. AB - BACKGROUND: The prevalence, features, and risk of cardiovascular disease (CVD) in dyslipidemic hypertension (DH) was investigated in a prospective population-based study. Dyslipidemic hypertension was defined in terms of blood pressure, plasma triglycerides, and HDL-cholesterol consistent with the metabolic syndrome criteria of the National Cholesterol Education Program guidelines. High-normal or hypertensive values not meeting the other two criteria were designated as "simple hypertensives" (SH). METHODS: A sample of 2225 men and women and free of CVD at baseline were followed up for a mean of 4.1 years. The proportions of DH, SH, and normotensives were 16%, 37%, and 47%, respectively. All persons with DH had metabolic syndrome by definition, whereas metabolic syndrome formed 44.6% of SH. Fatal and nonfatal CVD, diagnosed by clinical findings and Minnesota coding of resting electrocardiograms, developed in 166 subjects. RESULTS: Compared to SH, sex- and age-standardized individuals with DH had significantly higher body mass index, apolipoprotein B, fasting insulin, glucose, and C-reactive protein levels, had higher prevalence of impaired fasting glucose and metabolic syndrome. Cox regression analysis revealed a 1.57-fold higher (confidence interval 1.08-2.28) hazard ratio (HR) for CVD in DH than in SH, after adjustment for sex, age, LDL cholesterol, and smoking status. The sex- and age-adjusted HR of DH was furthermore 1.45-fold higher than the remaining subjects with metabolic syndrome (P = .096). Among persons with DH, age, presence of diabetes, and pulse pressure proved to be independent predictors for CVD. High LDL-cholesterol levels and fasting hyperinsulinemia were associated with borderline significantly elevated relative risks among dyslipidemic hypertensives. CONCLUSIONS: Dyslipidemic hypertension, prevailing in 1 of every 6 adults, implicates characteristic features, confers excess CVD risk compared to the remainder of hypertensives and carries half the attributable cardiovascular risk due to metabolic syndrome. PMID- 15797663 TI - Measuring arteriolar-to-venous ratio in retinal photography of patients with hypertension: development and application of a new semi-automated method. AB - BACKGROUND: The extent of arteriolar narrowing has been recognized as a major sign of end-organ damage in patients with hypertension, but most methods used for its evaluation are highly dependent on the observer. We describe a new semi automated method to measuring arteriolar-to-venous (A/V) ratio in retinal photography and present its application in the evaluation of patients with hypertension. METHODS: In a cross-sectional study, 58 patients with hypertension had retinographies taken and digitized to determine the vessel diameter by direct measurement (micrometric method) or by the new microdensitometric method. Sub pixel resolution was obtained via cubic spline fitting of the edge of vessel walls. For each pair of adjacent pixels, 10 intermediate points were generated in the perpendicular direction of the wall. Vessel widths were automatically extracted from this synthetic curve, with cut-points defined where the exterior wall position equals the double of mean noise along the slice. RESULTS: The intra and interobserver kappa statistics for the diagnosis of abnormal A/V ratio by the microdensitometric measurements were 0.93 (P < .0001) and 0.85 (P = .0005), respectively. Systolic blood pressure was inversely and significantly associated with the A/V ratio measured by the microdensitometric method. CONCLUSIONS: The microdensitometric method is reliable, is easy to operate, and captures the expected association between blood pressure and retinal vessels narrowing. Its performance in clinical practice and in the prediction of cardiovascular events should be confirmed in larger databases with retinographies. PMID- 15797664 TI - Renal norepinephrine spillover during infusion of nonesterified fatty acids. AB - BACKGROUND: Sympathetic activity and renal norepinephrine spillover are increased in obese individuals. We have reported that infusion of nonesterified fatty acids increases blood pressure in animals through stimulation of the sympathetic nervous system. METHODS: In this study, we assessed the effect of increasing circulating nonesterified fatty acids on systemic and renal norepinephrine kinetics in healthy adults by infusing fat emulsion and heparin for 4 h. (3)H norepinephrine was infused for 60 min before and again during the last hour of the fatty acid infusion to assess norepinephrine kinetics. Renal venous blood samples were obtained to calculate renal norepinephrine spillover. RESULTS: Nonesterified fatty acid levels increased threefold during the first hour and remained elevated throughout the study. Arterial and renal venous plasma norepinephrine levels fell by 15% and 20%, respectively, during the infusion (P < .05 for both). Kinetic analysis indicated that systemic release of norepinephrine into an extravascular compartment decreased from 11.6 +/- 1.1 to 10.0 +/- 1.3 nmol/min/m(2) (P = .067) and renal venous norepinephrine spillover decreased from 454 +/- 54 pmol/min (P = .055). CONCLUSIONS: These results indicate that nonesterified fatty acids do not have a direct stimulating effect on whole-body or renal sympathetic activity. It is possible that increased plasma levels of fatty acids serve as a signal to decrease sympathetic tone during the fasting state. PMID- 15797665 TI - Erectile dysfunction and its influence on quality of life in patients with essential hypertension. AB - Epidemiological studies indicated that erectile dysfunction (ED) is more frequent and their quality of life (QOL) is deteriorated in patients with essential hypertension as compared with normotensive subjects. Since both ED and hypertension influence QOL, it is not clear which malady has a more serious influence on QOL. We conducted a case control study to investigate incidence and impact of ED on QOL. In 102 patients aged 40 to 59 under treatment for hypertension (Hypertensive group) and in 245 age-matched normal subjects (Control group), QOL and ED were assessed by SF-36 and IIEF5. Criteria for presence of ED is IIEF5 score less than 12. Age adjusted incidence of ED in Hypertension group was higher than that in Control Group (odds ratio 1.7, P < .05). Multivariate logistic regression analysis showed hypertension was related only to a low score in general health, ED was related to low scores in physical, mental health, emotional, vitality, and general health. These data suggest that the influence of ED on QOL is more serious than hypertension in middle age men. PMID- 15797666 TI - A unifying pathway for essential hypertension. AB - We present the hypothesis that most cases of essential hypertension occur via two phases. The first phase is initiated by episodes of renal vasoconstriction induced by a hyperactive sympathetic nervous system, activation of the renin angiotensin system, or hyperuricemia resulting from diet or genetics. During this phase the hypertension is salt resistant and renin dependent, and the kidney normal. Over time, preglomerular vascular disease develops (arteriolosclerosis), associated with tubulointerstitial inflammation; this shifts the hypertension to a salt-sensitive, volume-dependent, and renal-dependent pathway. This pathway unites many of the previous hypotheses on the etiology of hypertension, and offers insights into ways to prevent, ameliorate, or cure the underlying process. PMID- 15797667 TI - Oxidative stress and profibrotic action of aldosterone. PMID- 15797668 TI - Reproducibility and reliability of the 13C-acetate breath test to measure gastric emptying of liquid meal in infants. AB - OBJECTIVE: This study assessed the noninvasive 13C-acetate breath test (13C-ABT) as a reliable and reproducible method to evaluate gastric emptying in infants METHODS: Gastric emptying was measured simultaneously by scintigraphy and 13C-ABT in 11 infants with clinical symptoms of gastroesophageal reflux to validate the method compared with the gold standard. Gastric emptying was also measured with 13C-ABT in a separate group of 14 healthy infants on 2 consecutive days to evaluate reproducibility of the method. Half-emptying times obtained with scintigraphy and 13C-ABT were correlated with Pearson's analysis. The variability of the half-emptying times obtained on 2 consecutive days was analyzed with paired t test and by the coefficient of variation as proposed by Bland and Altman. RESULTS: The mean emptying times obtained with scintigraphy and 13C-ABT were not different (89 +/- 27 min and 70 +/- 39 min, respectively, P = 0.22). Both methods correlated significantly (r = 0.75, P < 0.05). Half-emptying times measured on 2 consecutive days with 13C-ABT were 69 +/- 31 min and 68 +/- 30 min, respectively (mean difference = 1.4 +/- 12.4 min, P = 0.67). Intraindividual coefficient of variation was 6.3%. CONCLUSIONS: This study supports the 13C-ABT method as a useful tool to evaluate gastric emptying of liquids in healthy infants and in infants with gastroesophageal reflux in the field of research and in clinical evaluations. PMID- 15797669 TI - Social risk factors for hospital malnutrition. AB - OBJECTIVE: Disease severity is considered an important risk factor for malnutrition in hospitalized patients. We investigated the effect of social factors and disease parameters on the development of malnutrition. METHODS: Nutritional state was assessed by the Subjective Global Assessment in 794 consecutively admitted patients in two hospitals in Berlin (n = 493, university hospital; n = 301, district hospital). The influences of age, sex, social status, life habits, and disease parameters on nutritional state was analyzed with univariate and multivariate logistic regression methods. RESULTS: Malnutrition was diagnosed in 22% of patients. Its prevalence was significantly higher in patients with malignant than with benign diseases (odds ratio [OR] = 1.568, P < 0.05) and in patients with multiple prescriptions (OR = 1.154, P < 0.001), but no difference was found between medical or surgical patients. The strongest risk factors for malnutrition in univariate analysis were age older than 60 y (OR = 2.861, P < 0.001), living alone (OR = 1.769, P = 0.002), and achieving a lower level of education (OR = 1.589, P < 0.05). Therefore, multivariate analysis was performed after stratification for age. Demonstrated independent additional risk factors were polypharmacy (OR = 2.367, P < 0.001) and malignant disease (OR = 4.114, P < 0.001) in young patients and polypharmacy (OR = 1.109, P < 0.002) and living alone (OR = 1.830, P = 0.008) in patients 60 y and older. CONCLUSIONS: These data show that patients who are older, less educated, and live alone are at high risk of developing malnutrition. Thus special attention should be given to these risk groups when evaluating nutritional status and nutritional support. PMID- 15797671 TI - Effects of thickened beverages fortified with inulin on beverage acceptance, gastrointestinal function, and bone resorption in institutionalized adults. AB - OBJECTIVES: We wanted to develop thickened beverages that contain soluble fiber (inulin) with acceptable consistency, taste, and texture and to determine the effects of these beverages on bone resorption markers (to determine calcium retention), bowel frequency, and indicators of gastrointestinal function in institutionalized adults bound to wheelchairs. METHODS: A double-blind, 3-wk, cross-over study testing 13-g/d inulin-fortified versus isocaloric standard modified starch-thickened beverages was conducted in institutionalized adults who were bound to wheelchairs and had dysphagia or did not have dysphagia. Beverage acceptability, as assessed by discriminative and descriptive sensory testing, bowel frequency, fecal output, and laxative use, were determined by direct testing or by nursing charts. Bone resorption was measured by using the urinary excretion of fasting calcium and of cross-linked N-telopeptides of collagen. RESULTS: Sensory panelists were unable to detect a difference between beverages thickened with modified starch and those fortified with inulin. Few differences were found between the control and inulin-fortified beverages for sensory descriptors. No significant difference was found in frequency of bowel movements between treatments; however, weighted bowel movement frequency increased by 13% with inulin (P < 0.01), whereas enema and laxative administration decreased by 13% (P < 0.05). Bone resorption, as an indicator of calcium retention, remained unchanged. CONCLUSIONS: Inulin was incorporated into thickened beverages, with no decrease in acceptability; when consumed, perceived stool output increased in residents of long-term care facilities. PMID- 15797670 TI - Effects of creatine on thermoregulatory responses while exercising in the heat. AB - OBJECTIVE: We hypothesized that creatine supplementation would interfere with normal body fluid shifts that occur during exercise in a hot environment due to its osmotic effect intracellularly. This study examined the effects of acute creatine loading (20 g/d for 5 d) on the thermoregulatory response of the body during a bout of exercise at 39 degrees C. METHODS: Subjects (15 men and 1 woman) performed a cycle test of maximum oxygen consumption to determine the proper work rate for the heat-stress test (40 min at 55% maximum oxygen consumption at 39 degrees C) and were assigned to a creatine group (n = 8) or a placebo group (n = 8) in a double-blind fashion. Each group performed the heat-stress test on two separate occasions: before supplementation and after supplementation (20 g/d of creatine with Gatorade or Solka-floc plus Gatorade). Dependent variables included rectal temperature, mean skin temperature, mean body temperature, and perceived thermal sensation. RESULTS: Repeated measure analysis of variance showed a significant (P < or = 0.05) increase in body weight in the group supplemented with Gatorade. Core temperature was significantly lower after supplementation for both groups combined (before supplementation at 37.85 degrees C and after supplementation at 37.7 degrees C), with no difference between groups. A significant three-way interaction (group x trial x time) was also found for rectal temperature, with both groups having significantly lower rectal temperature after supplementation. Mean body and mean skin temperatures showed no differences. CONCLUSIONS: Short-term creatine supplementation (20 g/d for 5 d) did not have a negative effect on thermoregulatory responses during exercise at 39 degrees C. PMID- 15797672 TI - Effects of calcium pyruvate supplementation during training on body composition, exercise capacity, and metabolic responses to exercise. AB - OBJECTIVE: We evaluated the effects of calcium pyruvate supplementation during training on body composition and metabolic responses to exercise. METHODS: Twenty three untrained females were matched and assigned to ingest in a double blind and randomized manner either 5 g of calcium pyruvate (PYR) or a placebo (PL) twice daily for 30 d while participating in a supervised exercise program. Prior to and following supplementation, subjects had body composition determined via hydrodensiometry; performed a maximal cardiopulmonary exercise test; and performed a 45-min walk test at 70% of pre-training VO2 max in which fasting pre- and post exercise blood samples determined. RESULTS: No significant differences were observed between groups in energy intake or training volume. Univariate repeated measures ANOVA revealed that subjects in the PYR group gained less weight (PL 1.2 +/- 0.3, PYR 0.3 +/- 0.3 kg, P = 0.04), lost more fat (PL 1.1 +/- 0.5; PYR -0.4 +/- 0.5 kg, P = 0.03), and tended to lose a greater percentage of body fat (PL 1.0 +/- 0.7; PYR -0.65 +/- 0.6%, P = 0.07), with no differences observed in fat-free mass (PL 0.1 +/- 0.5; PYR 0.7 +/- 0.3 kg, P = 0.29). However, these changes were not significant when body composition data were analyzed by MANOVA (P = 0.16). There was some evidence that PYR may negate some of the beneficial effects of exercise on HDL values. No significant differences were observed between groups in maximal exercise responses or metabolic responses to submaximal walking. CONCLUSIONS: Results indicate that PYR supplementation during training does not significantly affect body composition or exercise performance and may negatively affect some blood lipid levels. PMID- 15797673 TI - Dietary intake in patients with asthma: a case control study. AB - OBJECTIVE: This case-controlled study investigated differences in dietary intake between a population with and without asthma. METHODS: A case-controlled study of the dietary intakes of subjects with asthma (n = 54, average age 23.59 +/- 9.6 y; group 1) and healthy subjects (n = 54, average age 27.07 +/- 7.2 y; group 2) using 3-d dietary intakes was undertaken. Fifty-four patients with asthma and healthy non-asthmatic subjects matched by age and sex were selected. All enrolled subjects (asthmatic and non-asthmatic) received instruction to record their daily dietary intake for 3 d including a weekend day. Pulmonary function tests were performed in asthmatic patients. RESULTS: Total energy intake and distribution of energy were normal in both groups (39.7% carbohydrates in group 1 and 41% in group 2, 19.4% proteins in group 1 and 22% in group 2, and 40.9% lipids in group 1 and 37% in group 2). A higher intake of vitamin A (642 +/- 399 microg/d) was detected in non-asthmatic than in asthmatic subjects (459 +/- 242 microg/d, P < 0.05). Daily intake of vitamin C was higher in non-asthmatic subjects (124 +/- 70 mg) than in asthmatic subjects (93.1 +/- 63.9 mg, P < 0.05). Vitamin E intake was higher in non-asthmatic subjects (7.3 +/- 1.2 mg) than in asthmatic subjects (5.1 +/- 2.3 mg, P < 0.05). Correlation analysis between daily intake and forced expiratory volume in 1 s (FEV1) in group 1 (asthmatic subjects) showed a positive association between omega-3 fatty acids (g/d) and FEV1 (r = 0.6, P < 0.05). No statistical correlations were detected between omega-6 and omega-9 fatty acid intakes and FEV1. Multivariant analysis with the dependent variable of FEV1 showed that only vitamin C and omega-3 fatty acids remained in the model after adjusting for smoking, age, and sex (F = 14.24, P < 0.001), with an increase of 0.25% (95% confidence interval = 0.1-0.4) in FEV1 with each 1 mug of vitamin C intake and an increase of 99.3% (95% confidence interval = 81.59-137.19) in FEV1 with each 1 g of omega-3 fatty acids. CONCLUSIONS: Asthmatic patients have a lower intake of vitamins A, C, and E than do non-asthmatic subjects. Intake of omega-3 fatty acids was correlated with FEV1. PMID- 15797674 TI - Are breast-fed infants and toddlers in New Zealand at risk of iodine deficiency? AB - OBJECTIVE: This study assessed the iodine status of New Zealand infants and toddlers and explored factors that might influence their iodine status. METHODS: A community-based, cross-sectional survey of 6- to 24-mo-old children was conducted in three cities in the South Island of New Zealand. Iodine status was determined by a casual urine sample. Breast-feeding mothers were asked to provide a breast milk sample for iodine determination. Caregivers collected a 3-d weighed diet record from their children to investigate associations between dietary patterns and urinary iodine excretion. RESULTS: The median urinary iodine concentration for the group (n = 230) was 67 microg/L (interquartile range 37 115) with 37% (95% confidence interval 30.5-43.4) of children having a urinary iodine concentration lower than 50 microg/L. When children were classified by current feeding method, those children who were currently formula-fed had a significantly higher median urinary iodine concentration (99 microg/L) than did children who were currently breast-fed (44 microg/L; P < 0.000). The mean iodine concentration in breast milk was 22 microg/L (n = 39). After multivariate analysis using estimates from 3-d diet records, only percentage of energy from infant formula was significantly associated with urinary iodine concentration (P = 0.005). CONCLUSIONS: This study found mild iodine deficiency in a group of New Zealand infants and toddlers. Children who consumed infant formula, which is fortified with iodine, had better iodine status than did children who were currently breast-fed because breast milk contained low levels of iodine. PMID- 15797675 TI - Predicting total urinary nitrogen excretion from urinary urea nitrogen excretion in multiple-trauma patients receiving specialized nutritional support. AB - OBJECTIVE: We investigated the accuracy of methods to estimate total urinary nitrogen (TUN) excretion from urinary urea nitrogen (UUN) excretion for patients who have multiple trauma and receive specialized nutritional support. METHODS: Fifty-five critically ill, adult patients who had multiple trauma and were receiving specialized nutritional support were evaluated. A 24-h urine collection for urea nitrogen and total nitrogen was performed 4.4 +/- 2.6 d after admission to the trauma intensive care unit. Patients with significant renal impairment, liver dysfunction, or obesity (>150% of ideal body weight) were excluded from study entry. Eight publications that examined the relation between TUN and UUN were evaluated for bias and precision in estimating TUN from UUN. RESULTS: TUN was 20.8 +/- 10.8 g/d with an average difference of 3.8 +/- 2.8 g/d between TUN and UUN. Linear regression analysis comparing TUN with UUN indicated a significant correlative relation (TUN = 1.1 x UUN + 2; r = 0.958, P < 0.001). The difference between TUN and UUN varied based on UUN: for UUN lower than 10 g/d, TUN minus UUN was 1.5 +/- 1.0 g/d; for UUN 10 to 20 g/d, TUN minus UUN was 4.1 +/ 3.2 g/d; and for UUN higher than 20 g/d, TUN minus UUN was 5.3 +/- 1.9 g/d (P < 0.001). Six methods were biased toward underpredicting TUN, one method was unbiased, and one was biased toward overpredicting TUN. A practical method for estimating TUN from UUN was developed: TUN = UUN + 2 for those with UUN lower than 10 g/d and TUN = 1.1 x UUN + 2 for those with UUN of at least 10 g/d. CONCLUSIONS: Our method, the modified Velasco method, UUN/0.84, and UUN/0.85 provided reasonable estimates of TUN from UUN in critically ill, adult patients who had multiple trauma and were receiving specialized nutritional support; however, our method requires further validation. PMID- 15797677 TI - Cardiac carnitine leakage is promoted by cardiomyopathy. AB - OBJECTIVE: We investigated whether a damaged heart with cardiomyopathy (CM) influences cardiac-stored carnitines. METHODS: A sensitive, specific, carnitine requiring yeast was used to determine blood carnitine concentration in 116 healthy subjects. For comparison with blood carnitine concentrations from patients with CM, we selected 33 male patients, ages 29 to 67 y, with evidence of CM and 24 male patients, ages 31 to 66 y, with no CM as categorized by cardiac catheterization. RESULTS: During catheterization, significantly higher concentrations of arterial blood levels of carnitines leaked from hearts of patients specifically with CM; no arterial blood carnitines leaked from hearts of patients without CM. Venous blood carnitine concentration for all patients was within the normal range. Carnitine did not accumulate in venous blood and was not a source of large amounts of leaked blood carnitines in patients with CM. CONCLUSION: CM causes leakage of carnitines from heart stores, possibly making cardiac tissue vulnerable to damage. We do not know whether cardiac carnitine leakage leads to CM or if established CM promotes cardiac carnitine leakage. PMID- 15797676 TI - Effect of the type of dietary fat on biliary lipid composition and bile lithogenicity in humans with cholesterol gallstone disease. AB - OBJECTIVE: The effect of the type of dietary fat on bile lipids and lithogenicity is unclear. This study compared the effects of two dietary oils that differed in fatty acid profile on biliary lipid composition in humans. METHODS: Female patients who had cholesterol gallstones and were scheduled for elective cholecystectomy were studied. For 30 d before surgery, subjects were kept on diets that contained olive oil (olive oil group, n = 9) or sunflower oil (sunflower oil group, n = 9) as the main source of fat. Gallbladder bile and stones were sampled at surgery. After cholecystectomy, duodenal samples were collected by nasoduodenal intubation during fasting and after administration of mixed liquid meals that included the corresponding dietary oil. Duodenal and gallbladder bile samples were analyzed for cholesterol, phospholipids, and total bile acids by established methods. Individual bile acid conjugates in gallbladder bile were measured by high-performance liquid chromatography. Gallstones were analyzed by semiquantitative polarizing light microscopy. RESULTS: Despite marked differences in the absolute concentration of biliary lipids and total lipid content, manipulation of dietary fat ingestion did not influence the cholesterol saturation or the profile of individual bile acids in gallbladder bile obtained from patients who had gallstones. All but one subject had mixed cholesterol stones. A cholesterol saturation index of hepatic bile in fasted cholecystectomized patients was similar in both dietary groups and indicative of supersaturation. In response to the test meal, the cholesterol saturation index decreased significantly in patients given the olive oil diet, reaching values lower than one at 120 min postprandially. In contrast, hepatic bile secreted by patients who consumed sunflower oil appeared supersaturated (cholesterol saturation index >1.5) throughout the experiment. CONCLUSIONS: Our results suggest that the type of dietary fat habitually consumed can influence bile composition in humans. In gallbladder, this influence was noted in the presence of more concentrated bile in the olive oil group. However, this was not translated into a modification of cholesterol saturation, which is likely due to the fact that cholesterol gallstones were present by the time the dietary intervention started. The finding that a typical postprandial variation in hepatic bile lithogenicity occurred only in olive oil patients was revealing. While keeping in mind the methodologic limitations of this part of the study, some gastrointestinal and metabolic mechanisms for this effect are discussed. PMID- 15797680 TI - Monounsaturated and omega-3 but not omega-6 polyunsaturated fatty acids improve hepatic fibrosis in hypercholesterolemic rabbits. AB - OBJECTIVE: Although the influence of saturated fatty acids, monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs), lipids, cholesterol levels, and other blood lipids has been established, few studies have examined the influence of these dietary lipids on the composition and histologic damage of organs in situations of hypercholesterolemia. Biliary lipids come from the liver, and this organ is essential in cholesterol homeostasis; thus, it may be helpful to evaluate the inter-relations among biliary, hepatic lipids, and hepatotoxic effects in situations of hypercholesterolemia with different dietary lipids. This study investigated whether administration of diets differing in fatty acid profiles (omega-3 PUFA, omega-6 PUFA, or MUFA) influence the content of biliary lipids, the lithogenic index of gallbladder bile, and the development of hepatic fibrosis in hypercholesterolemic rabbits. METHODS: Thirty rabbits were randomized to one of five groups. A control group received rabbit chow for 80 d. The remaining four groups received a 50-d diet that contained 3% lard and 13% cholesterol to provoke hypercholesterolemia. After this period, three groups were fed for another 30 d on a diet enriched with omega-6 PUFAs, MUFAs, and omega-3 PUFAs, respectively. Liver, bile, and plasma lipid compositions, lipid peroxidation in hepatic mitochondria, and histologic hepatic lesions were analyzed. RESULTS AND CONCLUSIONS: There was a beneficial effect of MUFA and omega-3 PUFA on hepatic fibrosis in hypercholesterolemic rabbits because both dietary fats led to recovery from hepatic lesions. However, because intake of omega-3 PUFA provoked lithogenic bile in rabbits, MUFA intake would be more advisable. PMID- 15797679 TI - Effects on immune response of antidiabetic ingredients from white-skinned sweet potato (Ipomoea batatas L.). AB - OBJECTIVES: The present report describes the effects of antidiabetic ingredients from white-skinned sweet potato (AWSSP) on the immune response of human cells. METHODS: We studied the effects of inactive Staphylococcus aureus cells coated with AWSSP on phagocytic activity, phagosome-lysosome fusion, and superoxide anion release by human leukocytes in vitro. RESULTS: AWSSP increased phagocytic activity and phagosome-lysosome fusion in neutrophils and monocytes in a dose dependent manner. In contrast, AWSSP had no significant effect on superoxide anion release (O2-) from human neutrophils. CONCLUSIONS: These results show that AWSSP is useful in the prevention and improvement of diabetic symptoms by stimulating human immunity and that Ipomoea batatas L. is a beneficial food because it increases immune activity in addition to its antidiabetic effects. PMID- 15797678 TI - Effect of selenium-induced oxidative stress on the cell kinetics in testis and reproductive ability of male mice. AB - OBJECTIVE: The present study evaluated the role of experimental oxidative stress (induced by feeding diets with different concentrations of selenium [Se], a trace nutrient and potent antioxidant) on male reproductive activity in mice. METHODS: To create different levels of oxidative stress in male mice, three diets with different levels of Se were fed to different groups for 8 wk. Mice in group 1 were fed a yeast-based diet, which is considered a Se-deficient diet (0.02 ppm). Mice in groups 2 and 3 were fed with an Se-deficient diet supplemented with 0.2 and 1 ppm Se as sodium selenite, respectively. RESULTS: After completion of the feeding schedule, a significant decrease in Se levels were observed in Se deficient mice (group 1), whereas Se levels greatly increased in the Se-excess mice (group 3). Glutathione peroxidase activity was greatly decreased in the liver and testis in group 1, whereas glutathione-S-transferase activity was significantly increased in the testis. No significant change was found in activities of glutathione peroxidase and glutathione-S-transferase in group 3 compared with group 2. Cell kinetics showed a significant decrease in the number of pachytene spermatocytes and young and mature spermatids in group 1 compared with group 2. No appreciable change was observed in the germinal cell population in group 3. A significant decrease in sperm number was observed in group 1 compared with group 2. No change in these parameters was observed in group 3. The fertility status of mice in terms of percent fertility and litter size also exhibited a significant decrease in the reproductive ability of group 1. No change in these parameters was observed in group 3 compared with group 2. CONCLUSION: The present results clearly demonstrate the effect of oxidative stress generated by feeding different concentrations of Se on cell kinetics in the testis and, hence, its effect on the reproductive ability of male mice. PMID- 15797682 TI - Caloric restriction modulates insulin receptor signaling in liver and skeletal muscle of rat. AB - OBJECTIVE: We investigated how the insulin/insulin-like growth factor-1 signaling pathway is involved in the robust antiaging effects produced by caloric restriction. METHODS: We subjected male rats to feeding ad libitum or calorie restriction, i.e., 60% of the ad libitum amount, for 2 and 25 mo and then assessed the effects of calorie restriction on insulin receptor (IR) signaling in liver and skeletal muscle. RESULTS: The results indicated that aging was accompanied by a significant decrease in IR tyrosine phosphorylation after insulin stimulation in live and skeletal muscle, which was associated with a significant increase in the activity of protein tyrosine phosphatase-1B. However, these age-related alterations were attenuated by long-term calorie restriction. Expression profile of mRNA showed an increased expression of mRNAs for IR and insulin-like growth factor-1 receptor in both tissues of calorie-restricted rats, but increased expression of IR mRNA was dissociated with the IR gene product in rats maintained on long-term calorie-restricted diet. CONCLUSION: IR signaling may play an important role in aging and its retardation by calorie restriction, and normal function of IR in liver and skeletal muscle is required for healthy aging and extending lifespan in mammals. PMID- 15797681 TI - High-methoxyl pectin has greater enhancing effect on glucose uptake in intestinal perfused rats. AB - OBJECTIVE: Pectins have been known to decrease blood glucose levels. However, the mechanism of this effect is unclear. The direct action of various pectins (high- or low-methoxyl pectins) on the intestinal absorption of glucose was investigated in gut-perfused rats. METHODS: After equilibrium, jejunal and ileal segments were simultaneously perfused with an isotonic electrolyte solution (pH 7.4) containing glucose (10 mM/L) and high- or low-methoxyl pectins (10 g/L). Each test or control solution was perfused in a random sequence, with perfusion times of 30 min. Changes in glucose concentration of perfusate solution reservoir were determined over the experimental period. RESULTS: High- and low-methoxyl pectins in the perfusate significantly inhibited jejunal uptake of glucose compared with the control (P < 0.05). High-methoxyl pectins had greater inhibitive effect on intestinal absorption of glucose than low-methoxyl pectins. The observed changes in glucose and water absorptions caused by high- or low-methoxyl pectins were reversible by switching to a pectin-free perfusate. In addition, net water absorption changed to secretion after addition of high- or low-methoxyl pectins. CONCLUSIONS: These results suggest that the decrease in intestinal absorption of glucose observed after perfusion of high- or low-methoxyl pectins may be caused by viscosity-related increases in mucosal unstirred layer thickness. PMID- 15797684 TI - Comparative toxicity of oleic acid and linoleic acid on Raji cells. AB - OBJECTIVE: Parenteral diets are often administered to critically ill patients. To study one of the effects of commercially available parenteral lipid diets, rich in triacylglycerol esters of omega-6 polyunsaturated fatty acids or omega-9 monounsaturated fatty acids, on the immune system of such patients, we evaluated the cytotoxicity of oleic and linoleic acids on Raji cells that had been derived from human B-lymphocytes. METHODS: Cell death intensity and type were investigated by flow cytometry by quantitation of cell volume, granularity, DNA fragmentation, mitochondrial depolarization, and lipid accumulation. Fluorescence microscopy was used to determine chromatin condensation and type of cell death (acridine orange/ethidium bromide assay). Gene expression of BCL-XL, BCL-XS, C MYC, and P53 was studied by reverse transcriptase polymerase chain reaction. RESULTS: Oleic acid was less toxic than linoleic acid to Raji cells. Both fatty acids promote apoptosis and necrosis of these cells. The mechanism of cell death induced by these fatty acids seemed to involve mitochondrial depolarization, lipid accumulation, and overexpression of C-MYC and P53. CONCLUSION: Oleic acid may offer a less harmful alternative to linoleic acid in parenteral diets with respect to patient B-lymphocyte-mediated immunologic activity. PMID- 15797683 TI - Differential anti-inflammatory effects of phenolic compounds from extra virgin olive oil identified in human whole blood cultures. AB - OBJECTIVE: The olive oil-rich Mediterranean diet protects against cardiovascular disease, which involves inflammatory processes. This study investigated the effects of phenolic compounds found in extra virgin olive oil on inflammatory mediator production by human mononuclear cells. METHODS: Diluted human blood cultures were stimulated with lipopolysaccharide in the presence of phenolics (vanillic, p-coumaric, syringic, homovanillic and caffeic acids, kaempferol, oleuropein glycoside, and tyrosol) at concentrations of 10(-7) to 10(-4) M. Concentrations of the inflammatory cytokines tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 and of the inflammatory eicosanoid prostaglandin E2 were measured by enzyme-linked immunosorbent assay. RESULTS: Oleuropein glycoside and caffeic acid decreased the concentration of interleukin 1beta. At a concentration of 10(-4) M, oleuropein glycoside inhibited interleukin 1beta production by 80%, whereas caffeic acid inhibited production by 40%. Kaempferol decreased the concentration of prostaglandin E2. At a concentration of 10(-4) M, kaempferol inhibited prostaglandin E2 production by 95%. No effects were seen on concentrations of interleukin-6 or tumor necrosis factor-alpha and there were no effects of the other phenolic compounds. CONCLUSIONS: Some, but not all, phenolic compounds derived from extra virgin olive oil decrease inflammatory mediator production by human whole blood cultures. This may contribute to the antiatherogenic properties ascribed to extra virgin olive oil. PMID- 15797685 TI - Effect of folic acid supplementation on aluminum accumulation in rats. AB - OBJECTIVE: Exposure to many xenobiotics may cause depletion of folic acid (folate), which is an essential vitamin for humans. Replacement of folate can be effective in protection against some diseases and in partial or total prevention of adverse effects related to xenobiotics. Aluminum (Al) is the most widely distributed metal in the outer crust of the earth. Its toxicity in humans is well known. However, there is no evidence that folate can decrease accumulation of Al to which humans can be exposed in many ways. The aim of the present study was to quantify organ Al accumulation and to evaluate whether there is any protective (or reductive) effect of folic acid on Al accumulation. METHODS: Male Wistar rats were assigned oral Al chloride (200 mg x kg(-1) x d(-1), n = 10, group 1) alone or in combination with folic acid (20 mg x kg(-1) x d(-1), n = 10, group 2) for 8 wk. At the end of the period, bone, kidney, brain, and blood samples were collected, and Al concentrations were determined by electrothermal atomic absorption spectrophotometry. RESULTS: Mean values of Al in the tissue samples from group 1 were higher than those from group 2 (all P < 0.05). No difference was observed in serum Al levels between groups (P > 0.05). CONCLUSION: These results suggest that folate supplementation might be useful to decrease Al accumulation in its main target organs, i.e., bone, kidney, and brain. PMID- 15797686 TI - Dietary fiber and body weight. AB - OBJECTIVE: This review provides an update of recent studies of dietary fiber and weight and includes a discussion of potential mechanisms of how dietary fiber can aid weight loss and weight maintenance. METHODS: Human studies published on dietary fiber and body weight were reviewed and summarized. Dietary fiber content of popular low-carbohydrate diets were calculated and are presented. RESULTS: Epidemiologic support that dietary fiber intake prevents obesity is strong. Fiber intake is inversely associated with body weight and body fat. In addition, fiber intake is inversely associated with body mass index at all levels of fat intake after adjusting for confounding factors. Results from intervention studies are more mixed, although the addition of dietary fiber generally decreases food intake and, hence, body weight. Many mechanisms have been suggested for how dietary fiber aids in weight management, including promoting satiation, decreasing absorption of macronutrients, and altering secretion of gut hormones. CONCLUSION: The average fiber intake of adults in the United States is less than half recommended levels and is lower still among those who follow currently popular low-carbohydrate diets, such as Atkins and South Beach. Increasing consumption of dietary fiber with fruits, vegetables, whole grains, and legumes across the life cycle is a critical step in stemming the epidemic of obesity found in developed countries. The addition of functional fiber to weight-loss diets should also be considered as a tool to improve success. PMID- 15797687 TI - The old enigma of diets, oils, and stones. PMID- 15797688 TI - Plant polyphenols: modifiers of immune function and risk of cardiovascular disease. PMID- 15797690 TI - Update on the treatment of bipolar disorder in children and adolescents. AB - As the phenomenology of pediatric bipolar disorder has become better delineated, clinicians are now able to more accurately assess and treat young people suffering from this condition. For pediatric patients with bipolar I disorder and symptoms of mania, medication monotherapy has been shown to lead to symptom amelioration. However, this treatment modality oftentimes does not lead to full symptom remission. In an attempt to address this observation, combination treatment strategies have recently been investigated. Recently, a maintenance study has shown that in youths who achieved remission on a combination of lithium and divalproate therapy, either of these agents alone was equally effective as a treatment strategy. In youths identified as being at genetic high risk for bipolarity who also had problematic affective symptomatology, treatment with divalproate was not found to be superior to placebo; however, those with the greatest degree of genetic risk for familial psychopathology remained in the trial longer than those with more modest amounts of familial psychopathology. These data suggest that intervention in youths with only one affected parent may not be a rational prevention strategy for pharmacological intervention in bipolar disorder, and that cohorts more genetically at risk may be a more appropriate group for preventative pharmacotherapy. PMID- 15797691 TI - New data on the use of lithium, divalproate, and lamotrigine in rapid cycling bipolar disorder. AB - The rapid cycling variant of bipolar disorder is defined as the occurrence of four periods of either manic or depressive illness within 12 months. Patients suffering from this variant of bipolar disorder have an unmet need for effective treatment. This review examines two major studies in an attempt to update understanding of the current therapies available to treat rapid cycling patients. The first trial compares lamotrigine versus placebo in 182 patients studied for 6 months. The second is a recently completed, 20-month trial comparing divalproate and lithium in 60 patients. Both trials had a double-blind, randomized parallel group design. The data from the latter study indicate that there are no large differences in efficacy between lithium and divalproate in the long-term treatment of rapid cycling bipolar disorder. In addition, lamotrigine has the potential to complement the spectrum of lithium and divalproate through its greater efficacy for depressive symptoms. PMID- 15797692 TI - The treatment of mixed states and the risk of switching to depression. AB - There are few controlled studies evaluating the treatment of bipolar mixed states. Evidence suggests that mixed states may be more responsive to some anticonvulsants than to lithium. Olanzapine alone or in combination with divalproate or lithium has been adequately evaluated in randomized clinical trials involving mixed-state patients, whereas risperidone and quetiapine have not. There is also some evidence demonstrating the efficacy of ziprasidone and aripiprazole. The risk of switching to depression is high in mixed states. Conventional antipsychotics, such as haloperidol, may be less efficacious at protecting against a switch to depression than atypical antipsychotics, divalproate or lithium. When choosing drugs for the treatment of mania, and especially for the treatment of mixed states, their efficacy against manic and depressive symptoms, and their safety in terms of the risk of switching to depression should be taken into account. PMID- 15797693 TI - Prediction of repeated parasuicide after 1-12 months. AB - To investigate predictors for repetition of suicide attempts 1-12 months after a suicide attempt. SUBJECTS AND METHODS: Two hundred and sixteen patients who had made a suicide attempt were investigated after 1 month, and 178 were followed up again after 12 months. RESULTS: During 1-12 months after the suicide attempt, 30 patients reattempted suicide (repeaters). During 0-1 month 13 patients had reattempted suicide (early repeaters), and nine of them also repeated between 1 and 12 months. Repeaters had more often made three or more attempts before index attempt, they more often were in treatment at the index attempt and at 1 month they had lower global functioning and higher suicide ideation. In a Cox Regression analysis two predictors for repetition between 1 and 12 months remained significant; early repetition (OR 6.7, 95% CI, 3.0-14.9) and having GAF scores below 49 (median cut-off) (OR 3.4 (95% CI, 1.5-7.5). CONCLUSION: Our findings suggest that repetitive behaviour in itself is a strong predictor of future attempts. Strategies focusing on the repetitive behaviour are warranted. PMID- 15797694 TI - Suicide in cyprus 1988-1999. AB - Suicide is a universally observed human behavior related to bio-psychological, social and cultural factors. The aim of the present study was to examine suicide in Cyprus, an island that has known many civilizations and cultures. All completed suicide cases in the Christian population of Cyprus during the years 1988-1999 were included in the study and they were analyzed according to age, gender, reported reasons for suicide and suicide methods. The main results indicate that: 1. The mean age-standardized suicide rate is the lowest in Europe, in males (3.08/100,000) and also in females (1.05/100,000). 2. Mean suicide rates increase significantly with age in males only. 3. Female suicide rates are highest in the 15-24 age group. 4. Statistically significant rising trends of male and female suicide rates in the all-ages group. 5. Suicide methods were mostly violent. Among males, the most common methods were poisoning, firearms explosives, and hanging, while in females, jumping, hanging and poisoning. 6. Mental disorders, physical illness, interpersonal and financial problems were the main reported reasons for suicide. The epidemiological characteristics of suicide in Cyprus might be attributed to a combined effect of social and cultural factors and probably reflect influences from countries to which Cyprus is ethnically, historically or geographically related. PMID- 15797695 TI - Long-term follow-up after severe suicide attempt by multiple blunt trauma. AB - OBJECTIVE: After parasuicide there is a high risk of reattempts. However, it seems that patients who survived severe suicidal trauma recover well. Therefore, the outcome of patients with severe multiple blunt trauma as a result of a suicide attempt was investigated with respect to psychiatric and somatic health, quality of life (QOL) and suicide reattempt rates. METHODS: Patients who underwent a suicide attempt were isolated from a prospectively collected sample of trauma patients from a level I University Trauma Centre. Follow-up examination was performed 6.1 +/- 3 years after the trauma. A physical and psychiatric examination was performed, using established psychiatric scales. RESULTS: Twelve percent of severely injured patients were identified as suicide attempters (male/female: 37/28, mean age 38 +/- 18 years, mean Injury Severity Score (ISS) 40 +/- 15 points). A psychiatric diagnosis was present in 90% at the time of the suicide attempt. Twenty-one patients died during the hospital stay (32%) and six subjects died thereafter, none due to suicide. Thirty-five individuals were eligible for examination. None of them had reattempted suicide. Seventeen (48%) had good outcomes reflected by absent or ambulatory psychiatric treatment, employment, normal psychiatric findings and good psychosocial ability. An indeterminate outcome was determined in 24%. Predictive variables for an adverse outcome (10 patients, 28%) were found to be a diagnosis of schizophrenia, continued psychiatric treatment and being without employment. CONCLUSION: Despite the seriousness of the suicide attempt, survivors recovered well in about half the cases with no further suicide attempt in any patient. An early psychiatric consultation already on the Intensive Care Unit (ICU) is recommended. PMID- 15797696 TI - Effect of a Suicide Prevention Centre for young people with suicidal behaviour in Copenhagen. AB - BACKGROUND: In the 1980s, suicide rates in Denmark were among the highest in the world. In 1992, a Suicide Prevention Centre was opened in Copenhagen with a 2 week programme of social and psychological treatment. The aim of the study was to evaluate the effect of the Suicide Prevention Centre. METHODS: In a quasi experimental study, 362 patients in the Suicide Prevention Centre and a parallel comparison group of 39 patients were interviewed with European Parasuicide Study Interviewer Schedule I (EPSIS I), which is a comprehensive interview including several validated scales. All patients were invited to follow-up interviews with EPSIS II and followed in the National Patients Register and the Cause of Death Register. RESULTS: At the 1-year follow-up, 59% of patients in the intervention group and 53% of patients in the comparison groups were interviewed with EPSIS II. The intervention group obtained a significantly greater improvement in Beck's Depression Inventory, Hopelessness Scale, Rosenberg's Self-Esteem Scale and CAGE score and a significantly lower repetition rate. DISCUSSION: Although the design cannot exclude selection bias, it seems likely that the improvement in the intervention group was facilitated by the treatment. PMID- 15797697 TI - Nicotine use in suicides: a case-control study. AB - PURPOSE: Despite of higher rates of substance-related disorders in psychiatric patients and suicides than in the general population, there is no clear specificity to the relationship between nicotine use and other psychiatric disorders for suicide risk. METHODS: One hundred and sixty-three suicides (mean age 49.8 +/- 19.3 years; 64.4% males; using psychological autopsy method) and 396 control persons (mean age 51.6 +/- 17.0 years; 55.8% males) were assessed with a standardised semi-structured interview including SCID-I and SCID-II (for DSM-IV). Suicides and controls were compared in terms of nicotine consumption and psychiatric disorders. Logistic regression was used to evaluate the interactions of tobacco consumption with psychiatric disorders. RESULTS: Suicides were significantly more often current smokers and heavy users of cigarettes (> 20 cigarettes per day; P < 0.001, each). Alcohol dependence, other axis I disorders than substance-related disorders, and cluster B personality disorder(s) remained independent predictors for suicide in both genders, current nicotine consumption only in men (OR = 2.6, 95% CI 1.3-5.2). DISCUSSION AND CONCLUSIONS: In males, but not in females, nicotine consumption contributed to risk of completed suicide after control for psychiatric disorders and has to be considered as independent risk factor for suicide. PMID- 15797698 TI - Earthquake-related psychological distress and associated factors 4 years after the Parnitha earthquake in Greece. AB - Exposure to earthquakes has been associated with psychological distress and in particular the development of post-traumatic stress disorder (PTSD). Earthquake related psychological distress can be longstanding. The present study involved 157 Greek survivors of the 1999 Parnitha earthquake assessed approximately 4 years after the earthquake. Assessments were based on the Traumatic Stress Symptom Checklist (TSSC). Using stringent calibrations for the estimation of symptom presence 25% of the survivors endorsed at least 5 and 12% at least 10 TSSC symptoms. Approximately 22% of the survivors reported subjective distress and 15% impaired adjustment due to their symptoms. Intensity of fear during the earthquake and participation in rescue operations related to greater post earthquake psychological distress. The results suggest that the psychological consequences of earthquakes can be serious and long-standing even when the magnitude of the earthquake is moderate. Psychological treatments that have been proven to reduce fear and PTSD symptoms need to be made available to the survivors. Such treatments may also increase the survivors' psychological preparedness and emotional resilience in view of future earthquakes. PMID- 15797699 TI - Validation of the Peritraumatic Dissociative Experiences Questionnaire self report version in two samples of French-speaking individuals exposed to trauma. AB - OBJECTIVE: Peritraumatic dissociation is a risk factor for developing PTSD. The Peritraumatic Dissociative Experiences Questionnaire (PDEQ) is a self-report inventory used to assess dissociation that occurred at the time of a trauma. The aim of this study was the validation the PDEQ in French. METHOD: Ninety French speaking traumatized victims presenting to the emergency department were recruited. They were administered the PDEQ shortly after exposure and others trauma-related measures 2 weeks and 1 month posttrauma. RESULTS: Principal components factor analyses suggested a single factor solution for the PDEQ. Significant correlations between the PDEQ and acute and posttraumatic stress symptoms indicated moderate to strong convergent validity. The PDEQ also showed satisfactory test-retest reliability and internal consistency. CONCLUSION: This study is the first one to investigate such detailed psychometric findings on the PDEQ. This confirms the unity of the concept of peritraumatic dissociation and the value of the PDEQ-French Version to assess it. PMID- 15797701 TI - Posttraumatic stress disorder and psychiatric co-morbidity: symptoms in a random sample of female Bosnian refugees. AB - OBJECTIVES: This study investigated psychological symptoms in Bosnian women 3-4 years after their arrival in Sweden. SUBJECTS AND METHODS: A simple random sample of 163 Bosnian women aged 19-59 was drawn from the Swedish populations register in 1996. The control group consisted of 392 Swedish-born women. Data were collected in face-to-face interviews. The Hopkins Symptom Checklist 25 (HSCL-25) and the Posttraumatic Symptom Scale (PTSS-10) were used to measure psychological symptoms of depression, anxiety, psychological distress, and posttraumatic stress disorder (PTSD). Unconditional logistic regression was used to estimate odds ratios (OR) for psychological symptoms after adjustment for age, country of birth, education, marital status, economic difficulties, social network, and feeling secure. RESULTS: The prevalence of symptoms of PTSD was 28.3% among the Bosnian women. Bosnian women had significantly higher risks of symptoms of depression, anxiety, and psychological distress than Swedish-born women. For depression the odds ratio was 9.50 among Bosnian women. CONCLUSION: Psychiatric community interventions need to target Bosnian refugee women. Awareness among health-care workers who encounter these women in a clinical setting should be improved. PMID- 15797700 TI - Effects of internal displacement and resettlement on the mental health of Turkish children and adolescents. AB - AIMS: To evaluate the effects of internal displacement and resettlement within Turkey on the emotional and behavioral profile of children, age 5-18 after controlling for possible confounding and demographic variables. METHOD: We conducted a national population survey using a self-weighted, equal probability sample. We compared the CBCL, TRF and YSR responses regarding children with (n = 1644) and without (n = 1855) experience of internal displacement. We examined the effects of gender, age, paternal employment, resettlement, urban residence and physical illness. RESULTS: The children and adolescents with internal displacement had significantly higher internalizing, externalizing and total problem scores on the CBCL and YSR, and higher internalizing scores on the TRF. The effect of displacement was related to higher internalizing problems when factors like physical illness, child age, child gender and urban residence were accounted. The overall effect was small explaining only 0.1-1.5% of the total variance by parent reports, and not evident by teacher reports. DISCUSSION: To our knowledge the present study is the first to examine Turkish children and adolescents with and without experience of internal displacement. The results are consistent with previous immigration studies: child age, gender, presence of physical illness and urban residence were more important predictors of internalization and externalization problem scores irrespective of informant source. PMID- 15797702 TI - Predictability of a favorable outcome in anorexia nervosa. AB - OBJECTIVE: In a long-term follow-up of anorexia nervosa (AN) patients, somatic, psychological and social variables at clinical presentation should be investigated using a multilevel approach. METHODS: This study isolated predictors known from the literature over longer time periods and carried out a separate investigation of predictors in a sample of 81 AN patients of the Heidelberg Mannheim study over a mean period of 12 years (range 9-19 years). Separate hierarchic regression analyses on the basis of the course of the Morgan-Russell categories were calculated for four individually recorded areas: anamnestic, psychological, somatic and social data sets. RESULTS: Age at the onset of the disease, purging behavior, low serum albumin, high glutamic-oxalo acetic transaminase (GOT) psychopathology (ANSS) and social pathology had the highest predictive value qualities. In survival analysis overall assessment of all six main predictors at clinical presentation could differentiate all patients who recovered from those who remained ill (log-rank test P = 0.019). CONCLUSION: A small number of variables were important for detecting a good or poor long-term course of AN. At onset of the disease, it seems necessary to evaluate these psychological, somatic and social predictors. PMID- 15797703 TI - New contributions to the prevalence of eating disorders in Spanish adolescents: detection of false negatives. AB - BACKGROUND: Although the epidemiology of Eating Disorders (ED) has been highly developed in Spain, further research considering false negatives and also the prevalence of these disorders in males are needed. They were the aims of the present study. METHODS: One thousand and seventy six adolescents (500 males and 576 females) participated in a two-stage survey. At the age of 13, apart from exploring numerous clinical and psychosocial variables, they were assessed by means of self-administered screening instruments, the eating attitude test (EAT), bulimic investigatory test Edinburgh (BITE), and body shape questionnaire (BSQ). The subjects were interviewed again 2 years later. Afterwards, 159 probable cases and 150 controls were interviewed by means of the structured interview SCAN. RESULTS: Prevalence of ED in adolescents was 3.71% (95% CI 2.58-4.83; 6.4% of females, 0.6% of males): anorexia nervosa 0.1%, bulimia nervosa 0.75%, eating disorder not otherwise specified (EDNOS) = 2.88%. The rate of false negative was 2.6% of the control group and most of them were EDNOS. CONCLUSIONS: The prevalence of ED obtained in our study, higher than others found in previous Spanish research, and a relatively high percentage of false negatives both indicate that up to now the prevalence of ED in Spanish adolescents could be underestimated. PMID- 15797704 TI - Incidence of eating disorders in Navarra (Spain). AB - PURPOSE: To estimate the overall annual incidence and age group distribution of eating disorders in a representative sample of adolescent female residents of Navarra, Spain. METHODS: We studied a representative sample of 2734 adolescent Navarran females between 13 and 22 years of age who were free of any eating disorder at the start of our study. Eighteen months into the study, we visited the established centers and the eating attitudes test (EAT-40) and eating disorder inventory (EDI) Questionnaires were administered to the entire study population. We obtained a final response of 92%. All adolescents whose EAT score was over 21 points and a randomized sample of those who scored 21 or below, were interviewed. Any person meeting the DSM-IV diagnostic criteria for Anorexia Nervosa (AN), Bulimia Nervosa (BN) or eating disorder not otherwise specified (EDNOS) was considered a case. RESULTS: We detected 90 new cases of eating disorders. Taking into consideration the randomly selected group whose EAT score was 21 points or below, we estimated the overall weighted incidence of eating disorders to be 4.8% (95% CI: 2.8-6.8), after 18 months of observation, in which EDNOS predominated with an incidence of 4.2% (95% CI: 2.0-6.3). The incidence of AN was 0.3% (95% CI: 0.2-0.5), while that of BN was also found to be 0.3% (95% CI: 0.2-0.5). The highest incidence was observed in the group of adolescents between 15 and 16 years of age. CONCLUSIONS: The overall incidence of ED in a cohort of 2509 adolescents after 18 months of follow-up was 4.8% (95% CI: 2.8 6.8), with EDNOS outweighing the other diagnoses. The majority of new cases of eating disorders were diagnosed between ages 15 and 16. PMID- 15797705 TI - Modified nutritional counselling to increase motivation to treatment in anorexia nervosa. PMID- 15797706 TI - Temperament and character in primary insomnia. AB - Recent studies by Cloninger suggest that the temperament dimension of harm avoidance might be related to serotonergic activity. Since serotonergic mechanisms equally play a major role in sleep regulation, we decided to use Cloninger's psychobiological model of temperament and character to assess whether there is a link between psychophysiologic insomnia and specific personality traits. Chronic insomnia is a common complaint in modern society, and it is still controversial whether insomniacs share specific personality traits. Thirty-two chronic insomniacs (< 50 years) were studied. They underwent polysomnography for two consecutive nights and filled out the 226-item self-questionnaire of Temperament and Character Inventory as well as the Hospital Anxiety and Depression scale. (1) Harm avoidance for all subscores was significantly higher in insomniac patients when compared with controls; (2) self-directedness scores were lower in insomniacs; (3) sleep latency was positively correlated to harm avoidance; (4) HA1 (anticipatory worry) was negatively correlated to REM latency. Temperament and Character Inventory is a useful tool in the investigation of chronic insomnia. Serotonergic mechanisms might explain the high incidence of harm avoidance as personality trait in psychophysiologic insomniac patients. Further studies are needed to see whether harm avoidance could be a psychological vulnerability marker for primary insomnia and be used as predictor of SSRI treatment responders. PMID- 15797707 TI - Moroccan colloquial Arabic version of the Mini International Neuropsychiatric Interview (MINI): qualitative and quantitative validation. AB - The validation of mini international neuropsychiatric interview (MINI) into Moroccan Colloquial Arabic language demonstrated good psychometric properties. The concordance between translated MINI's and expert diagnoses was good with kappa values greater than 0.80. The reliability inter-rater and test-retest were excellent with kappa values above 0.80 and 0.90, respectively. PMID- 15797708 TI - Mouse neuron navigator 1, a novel microtubule-associated protein involved in neuronal migration. AB - The development of the nervous system (NS) requires the coordinated migration of multiple waves of neurons and subsequent processes of neurite maturation, both involving selective guidance mechanisms. In Caenorhabditis elegans, unc-53 codes for a new multidomain protein involved in the directional migration of a subset of cells. We describe here the first functional characterization of the mouse homologue, mouse Neuron navigator 1 (mNAV1), whose expression is largely restricted to the NS during development. EGFP-mNAV1 associates with microtubules (MTs) plus ends present in the growth cone through a new microtubule-binding (MTB) domain. Moreover, its overexpression in transfected cells leads to MT bundling. The abolition of mNAV1 causes loss of directionality in the leading processes of pontine-migrating cells, providing evidence for a role of mNAV1 in mediating Netrin-1-induced directional migration. PMID- 15797709 TI - Subcellular targeting of DISC1 is dependent on a domain independent from the Nudel binding site. AB - Disrupted in schizophrenia 1 (DISC1) has been identified as a putative risk factor for schizophrenia and affective disorders through study of a Scottish family with a balanced (1;11) (q42.1;q14.3) translocation, which results in the disruption of the DISC1 locus and cosegregates with major psychiatric disease. Several other reports of genetic linkage and association between DISC1 and schizophrenia in a range of patient populations have added credibility to the DISC1-schizophrenia theory, but the function of the DISC1 protein is still poorly understood. Recent studies have suggested that DISC1 plays a role in neuronal outgrowth, possibly through reported interactions with the molecules Nudel and FEZ1. Here we have analyzed the DISC1 protein sequence to identify previously unknown regions that are important for the correct targeting of the protein and conducted imaging studies to identify DISC1 subcellular location. We have identified a central coiled-coil region and show it is critical for the subcellular targeting of DISC1. This domain is independent from the C-terminal Nudel binding domain highlighting the multidomain nature/functionality of the DISC1 protein. Furthermore, we have been able to provide the first direct evidence that DISC1 is localized to mitochondria in cultured cortical neurons that are dependent on an intact cytoskeleton. Surprisingly, Nudel is seen to differentially associate with mitochondrial markers in comparison to DISC1. Disruption of the cytoskeleton results in colocalization of Nudel and mitochondrial markers-the first observation of such a direct relationship. Mitochondrial dysfunction has been implicated to play a role in schizophrenia so we speculate that mutations in DISC1 or Nudel may impair mitochondrial transport or function, initiating a cascade of events culminating in psychiatric illness. PMID- 15797710 TI - Receptor protein tyrosine phosphatase sigma inhibits axon regrowth in the adult injured CNS. AB - Recently, receptor protein tyrosine phosphatase-sigma (RPTPsigma) has been shown to inhibit axon regeneration in injured peripheral nerves. Unlike the peripheral nervous system (PNS), central nervous system (CNS) neurons fail to regenerate their axons after injury or in disease. In order to assess the role of RPTPsigma in CNS regeneration, we used the retinocollicular system of adult mice lacking RPTPsigma to evaluate retinal ganglion cell (RGC) axon regrowth after optic nerve lesion. Quantitative analysis demonstrated a significant increase in the number of RGC axons that crossed the glial scar and extended distally in optic nerves from RPTPsigma (-/-) mice compared to wild-type littermate controls. Although we found that RPTPsigma is expressed by adult RGCs in wild-type mice, the retinas and optic nerves of adult RPTPsigma (-/-) mice showed no histological defects. Furthermore, the time-course of RGC death after nerve lesion was not different between knockout and wild-type animals. Thus, enhanced axon regrowth in the absence of RPTPsigma could not be attributed to developmental defects or increased neuronal survival. Finally, we show constitutively elevated activity of mitogen-activated protein kinase (MAPK) and Akt kinase in adult RPTPsigma (-/-) mice retinas, suggesting that these signaling pathways may contribute to promoting RGC axon regrowth following traumatic nerve injury. Our results support a model in which RPTPsigma inhibits axon regeneration in the adult injured CNS. PMID- 15797711 TI - CAP-1A is a novel linker that binds clathrin and the voltage-gated sodium channel Na(v)1.8. AB - The voltage-gated sodium channel Na(v)1.8 produces a tetrodotoxin-resistant current and plays a key role in nociception. Annexin II/p11 binds to Na(v)1.8 and facilitates insertion of the channel within the cell membrane. However, the mechanisms responsible for removal of specific channels from the cell membrane have not been studied. We have identified a novel protein, clathrin-associated protein-1A (CAP-1A), which contains distinct domains that bind Na(v)1.8 and clathrin. CAP-1A is abundantly expressed in DRG neurons and colocalizes with Na(v)1.8 and can form a multiprotein complex with Na(v)1.8 and clathrin. Coexpression of CAP-1A and Na(v)1.8 in DRG neurons reduces Na(v)1.8 current density by approximately 50% without affecting the endogenous or recombinant tetrodotoxin-sensitive currents. This effect of CAP-1A is blocked by bafilomycin A1 treatment of transfected DRG neurons. CAP-1A thus is the first example of an adapter protein that links clathrin and a sodium channel and may regulate Na(v)1.8 channel density at the cell surface. PMID- 15797713 TI - Novel clustering of sodium channel Na(v)1.1 with ankyrin-G and neurofascin at discrete sites in the inner plexiform layer of the retina. AB - Voltage-gated sodium channels cluster at sites of action potential generation and propagation by interacting with partner proteins such as neurofascin, an adhesion molecule in the L1 family, and ankyrin-G, a spectrin-binding protein required for sodium channel accumulation at axon initial segments. Here, we describe in the inner plexiform layer of the retina a novel site of high-density sodium channel clustering, marked by ankyrin-G and neurofascin. The sodium channel isoform at this site is Na(v)1.1, instead of the Na(v)1.6 channels more commonly found in association with the clustering machinery. During development, Na(v)1.2 channels first associate with ankyrin-G in the inner plexiform layer but are later replaced by Na(v)1.1, similar to the switch from Na(v)1.2 to Na(v)1.6 at nodes of Ranvier and initial segments. This represents the first instance of high-density clustering of Na(v)1.1 channels, which may contribute to synaptic interactions among retinal neurons in the inner plexiform layer. PMID- 15797712 TI - NSF interaction is important for direct insertion of GluR2 at synaptic sites. AB - Here, we use a cell surface thrombin cleavage assay to investigate directly the role of NSF in the surface delivery and synaptic accumulation of alpha-amino-3 hydroxy-5-methylisoxazolepropionate (AMPA) receptors. In cultured hippocampal neurons, the GluR2 subunit (which specifically interacts with NSF) inserts rapidly into the plasma membrane from intracellular compartments and accumulates in synaptic sites. In contrast, surface accumulation of GluR3 (a subunit that does not interact with NSF) or a GluR2 mutant defective in NSF binding (DeltaA849 Q853) occurs initially at extrasynaptic sites and is kinetically slower than wild type GluR2. Introducing a binding site for NSF into GluR3 (GluR3NSF) generates a subunit that behaves like GluR2 in terms of kinetics and site of surface insertion. These data suggest that the NSF interaction is necessary for rapid incorporation of AMPA receptor subunits into synapses and is sufficient to confer this property on GluR3. PMID- 15797714 TI - Heterotypic neuronal differentiation of adult subependymal zone neuronal progenitor cells transplanted to the adult hippocampus. AB - Neuronal progenitor cells (NPCs) residing in the adult subependymal zone (SEZ) are a potential source of expandable cells for autologous transplantation to replace neurons lost in multiple types of brain injury. To characterize the capacity of these cells for neuronal differentiation in a mature ectopic environment, NPCs expanded from the SEZ of adult rats were transplanted to the adult dentate gyrus. Cultures comprised a heterogeneous population of proliferating cells, which expressed nestin (47%) and GFAP (37%), with many cells expressing both progenitor cell markers (31%). In grafts of undifferentiated cells, as well as in grafts of cells that were induced to differentiate in vitro with retinoic acid, 35% of the transplanted SEZ-derived cells exhibited immunohistochemical and morphological features characteristic of hippocampal granule cell neurons. These novel results indicate that in vitro expanded adult SEZ NPCs are capable of heterotypic neuronal differentiation in a neurogenic region of the adult brain. PMID- 15797715 TI - Characterization of the NF2 protein merlin and the ERM protein ezrin in human, rat, and mouse central nervous system. AB - The neurofibromatosis 2 (NF2) protein, merlin, is structurally related to the ERM (ezrin-radixin-moesin) protein family of membrane-cytoskeleton linkers and is mutated in nervous system tumors. Apart from tumor suppressor activity, merlin's functions are poorly understood. We compared the localization and expression of merlin and ezrin in developing and adult brain and in brain-derived progenitor cells. Both proteins were widely but differentially expressed in human, rat, and mouse brain. In brain tissue and neuronal progenitor cell cultures merlin was predominantly found in neurons while ezrin was expressed in astrocytes. Merlin expression was seen from E11 in mouse embryos, whereas ezrin was present earlier. Both proteins were expressed in embryonic mouse neurospheres, where ezrin was specifically localized in filopodia of adherent neuronal progenitor cells. Subcellular analysis demonstrated ezrin in fine filopodial structures in astrocytes, while merlin was detected in neuronal synaptic junctions. The widespread expression of merlin in brain and its association with protein kinase A suggest a role for merlin in brain biology. PMID- 15797716 TI - Accumulation of Nav1 mRNAs at differentiating postsynaptic sites in rat soleus muscles. AB - Acetylcholine receptors (AChRs) and voltage-gated sodium channels (Na(V)1s) accumulate at different times in the development of the murine neuromuscular junction (NMJ). We used in situ hybridization to study the relationship of Na(V)1 mRNA accumulation to this difference. mRNAs encoding both muscle Na(V)1 isoforms, Na(v)1.4 and Na(v)1.5, were first concentrated at NMJs at birth, when the proteins start to accumulate. Within 4 weeks, Na(v)1.4 mRNA increased 5-fold at the NMJ while Na(v)1.5 mRNA became undetectable. Na(V)1 mRNA accumulation occurred even if the nerve was cut at birth. Like AChR mRNA, Na(V)1 mRNA accumulated at denervated synaptic sites on regenerating muscles and in response to ectopically expressed neural agrin. Clustering of Na(V)1 at the NMJ follows that of its mRNA while AChR clustering precedes its mRNA clustering by several days. This suggests that factors other than local mRNA upregulation determine the timing of clustering of these two important postsynaptic ion channels. PMID- 15797718 TI - A novel function for cadherin-11 in the regulation of motor axon elongation and fasciculation. AB - We previously observed that cadherin-11, a type II cadherin, is expressed in growing motor and sensory axons in the mouse embryo. Here, we assessed its functional involvement in the regulation of axon elongation and fasciculation by evaluating the activity of a specific cadherin-11 homophilic ligand, cad11-Fc (cadherin-11 extracellular region fused to Fc fragment of IgG), on the length and organization of motor axons outgrowing from embryonic ventral spinal cord explants. Cad11-Fc substrate enhanced axon growth and prevented interactions occurring between growing axons, providing evidences for a role of cadherin-11 in the control of growth cone progression. Comparison of cadherin-11 with N cadherin, a type I cadherin concomitantly expressed by motor axons, revealed similarities in their functional properties, including the ability to reorganize the actin cytoskeleton through interactions with catenins, but differences in their axon growth-promoting activity, arguing for subtle differences in their contributions to peripheral nerve elongation. PMID- 15797717 TI - Experimental Charcot-Marie-Tooth type 1A: a cDNA microarrays analysis. AB - To reveal the spectrum of genes that are modulated in Charcot-Marie-Tooth neuropathy type 1A (CMT1A), which is due to overexpression of the gene coding for the peripheral myelin protein 22 (pmp22), we performed a cDNA microarray experiment with cDNA from sciatic nerves of a rat model of the disease. In homozygous pmp22 overexpressing animals, we found a significant down-regulation of 86 genes, while only 23 known genes were up-regulated, suggesting that the increased dosage of pmp22 induces a general down-regulation of gene expression in peripheral nerve tissue. Classification of the modulated genes into functional categories leads to the identification of some pathways altered by overexpression of pmp22. In particular, a selective down-regulation of the ciliary neurotrophic factor transcript and of genes coding for proteins involved in cell cycle regulation, for cytoskeletal components and for proteins of the extracellular matrix, was observed. Cntf expression was further studied by real-time PCR and ELISA technique in pmp22 transgenic sciatic nerves, human CMT1A sural nerve biopsies, and primary cultures of transgenic Schwann cells. According to the results of cDNA microarray analysis, a down-regulation of cntf, both at the mRNA and protein level, was found in all the conditions tested. These results are relevant to reveal the molecular function of PMP22 and the pathogenic mechanism of CMT1A. In particular, finding a specific reduction of cntf expression in CMT1A Schwann cells suggests that overexpression of pmp22 significantly affects the ability of Schwann cells to offer a trophic support to the axon, which could be a factor, among other, responsible for the development of axonal atrophy in human and experimental CMT1A. PMID- 15797719 TI - Context-dependent regulation of NeuroD activity and protein accumulation. AB - NeuroD/BETA2 (referred to as NeuroD hereafter) is a basic helix-loop-helix (bHLH) transcription factor that is required for the development and survival of a subset of neurons and pancreatic endocrine cells in mice. Gain-of-function analyses demonstrated that NeuroD can (i) convert epidermal fate into neuronal fate when overexpressed in Xenopus embryos, and (ii) activate the insulin promoter in pancreatic beta cell lines in response to glucose stimulation. In glucose-stimulated INS-1 pancreatic beta cells, mutations of S259, S266, and S274 to alanines inhibited the ability of NeuroD to activate the insulin promoter. Phosphorylation of those serine residues by ERK1/2 was required for NeuroD activity in that assay. To determine whether the same residues are implicated in the neurogenic activity of NeuroD, we mutated the conserved S259, S266, and S274 of Xenopus NeuroD to alanines (S259A, S266A, and S274A), and performed an ectopic neurogenesis assay in Xenopus embryos. In contrast to what has been observed in the pancreatic beta cell line, the S266A and S274A mutant forms of Xenopus NeuroD displayed significantly increased abilities to form ectopic neurons, while S259A had little effect. In addition, S266A and S274A of Xenopus NeuroD resulted in increased accumulation of protein in the injected embryos while the corresponding mutations on mouse NeuroD did not have the same effect in an insulinoma cell line. Our results demonstrate that the consequence of NeuroD protein modification is context-dependent at both the molecular and functional levels. PMID- 15797720 TI - Syntenin is involved in the developmental regulation of neuronal membrane architecture. AB - Syntenin is a approximately 33 kDa scaffolding protein that we have shown previously to bind to kainate receptor subunits via a PDZ interaction. Here we show that syntenin has a tightly regulated developmental profile in neurons and is most abundant in the period of intense growth and synapse formation and stabilization. There is extensive colocalization of syntenin and kainate receptors with particularly intense labeling for both proteins at growth cones. Overexpression of GFP-syntenin in both young and mature neurons evokes marked changes in neuronal morphology by increasing the number of dendritic protrusions. These results are consistent with the involvement of syntenin in controlling membrane organization and suggest that by interaction with kainate receptors it may play a role in determining the formation and maturation of synapses. PMID- 15797721 TI - Multiple motifs regulate the trafficking of GABA(B) receptors at distinct checkpoints within the secretory pathway. AB - gamma-Aminobutyric acid type B receptors (GABA(B)) are G-protein-coupled receptors that mediate GABAergic inhibition in the brain. Their functional expression is dependent upon the formation of heterodimers between GABA(B)R1 and GABA(B)R2 subunits, a process that occurs within the endoplasmic reticulum (ER). However, the mechanisms that regulate receptor surface expression remain largely unknown. Here, we demonstrate that access to the cell surface for GABA(B)R1 is sequentially controlled by an RSR(R) motif and a LL motif within its cytoplasmic domain. In addition, we reveal that msec7-1, a guanine-nucleotide-exchange factor (GEF) for the ADP-ribosylation factor (ARF) family of GTPases, critical regulators of vesicular membrane trafficking, interacts with GABA(B)R1 via the LL motif in this subunit. Finally, we establish that msec7-1 modulates the cell surface expression of GABA(B) receptors, a process that is dependent upon the integrity of the LL motif in GABA(B)R1. Together, our results demonstrate that the cell surface expression of the GABA(B)R1 subunit is regulated by multiple motifs, which act at distinct checkpoints in the secretory pathway, and also suggest a novel role for msec7-1 in regulating the membrane trafficking of GABA(B)R1 subunits. PMID- 15797723 TI - The presence of glia stimulates the appearance of glycinergic synaptic transmission in spinal cord neurons. AB - Previous studies, using electrophysiological and fluorimetric analysis with a calcium sensitive dye, have shown that 5-7 DIV developing spinal cord neurons displayed high levels of glycinergic transmission. GABAergic and AMPAergic neurotransmission contributed much less to the overall transmission. Here, we show that culturing neurons in absence of a glia cell monolayer reduced the frequency of glycinergic spontaneous IPSCs (0.1 +/- 0.01 Hz), without altering the level of overall transmission (3 +/- 1.1 Hz). The predominant transmission was mediated by GABA(A) receptors (72% of total synaptic events). In addition, combination of bicuculline and CNQX blocked synaptically mediated calcium transients recorded with fluo-3. Furthermore, application of glycine revealed the presence of extrasynaptic receptors in these neurons (25 +/- 6 pA/pF). Culturing neurons on a glial cell monolayer increased the frequency of glycinergic currents (0.4 +/- 0.02 Hz), without changing the amplitude of the current (20 +/- 4 pA). The use of a glia-conditioned media reversed the effect of growing the neurons in a glia-deprived condition. These results indicate that the establishment of glycinergic transmission is dependent on the presence of a glia derived soluble factor. However, functional GlyRs were still able to insert in the neuronal membrane in a glia-independent manner. PMID- 15797722 TI - Disruption of the mouse Large gene in the enr and myd mutants results in nerve, muscle, and neuromuscular junction defects. AB - The autosomal recessive neuromuscular disorder associated with the enervated (enr) mouse transgene insertion manifests impaired peripheral nerve regeneration due to defects in Schwann cells and resembles the myodystrophy (Large(myd)) phenotype. Here we show that the enr transgene has integrated into Chr 8 approximately 160 kb downstream from the 3' end of the Large gene disrupting its expression as confirmed by the lack of genetic complementation between Large(myd) and enr mice, the very low Large mRNA levels in enr tissues and hypoglycosylation of alpha-dystroglycan, a known substrate of LARGE. Mutant nerve conduction and grip strength were impaired whereas sodium channel clustering at the nodes of Ranvier was unaffected. Interestingly, the mutant neuromuscular junctions displayed abnormal acetylcholine receptor clustering with reduced immunostaining for beta-dystroglycan, laminin, agrin, MuSK, and to a lesser extent acetylcholinesterase and rapsyn. These data implicate LARGE in nerve, muscle, and neuromuscular junction function. PMID- 15797724 TI - Opposing roles of ERK and p38 MAP kinases in FGF2-induced astroglial process extension. AB - The stellate processes of astroglial cells undergo extensive remodeling in response to neural injury. Little is known about intracellular signaling mechanisms controlling process extension. We tested roles for the ERK and p38 MAP kinase pathways in a simplified culture model. FGF2-induced process extension was preceded by a strong and transient phosphorylation of ERK, and a modest activation of p38 MAP kinase, which exhibited significant basal activity. Phosphorylated ERK was found predominantly in the cytoplasm, whereas activated p38 MAP kinase was nuclear. Process extension was completely blocked by the specific MEK inhibitor U0126. Conversely, inhibition of the p38 MAP kinase pathway with SB202190 stimulated spontaneous process growth and greatly potentiated FGF2-induced process extension. The p38 inhibitor effect was reproduced with an adenovirus expressing dominant-negative p38 MAP kinase. Selective pharmacological blockade of MAP kinase pathways may enable modulation of the astroglial response to injury so as to promote neural regeneration. PMID- 15797725 TI - Novel type of enzyme multimerization enhances substrate affinity of oat beta glucosidase. AB - Oat beta-glucosidase in plastid hydrolyzes avenacosides to C26-desgluco avenacosides to combat against fungal infections. The enzyme has a unique quaternary protein structure of a three-dimensionally radiated assembly of long fibrillae. We elucidated the fibrillar assembly of oat type 1 beta-glucosidase by means of cryo-electron microscopy, enzyme kinetics and chemical modification. It was assembled by linear stacking of hollow trimeric units and the resulting fibril had a long central tunnel connecting to the outer medium via regularly distributed side fenestrations. The enzyme active sites were located within the central tunnel. This unique multimer assembly increased enzyme affinity to avenacosides, in vivo substrates, and may function to discriminate avenacosides from many other kinds of beta-glucoside in oat. The fibrillar multimer of oat beta-glucosidase is a novel quaternary protein structure for enzyme supramolecular assembly that may have a functional role in the regulation of enzyme affinity. PMID- 15797726 TI - Crystal structure of the tryparedoxin peroxidase from the human parasite Trypanosoma cruzi. AB - Tryparedoxin peroxidase from Trypanosoma cruzi (TcTXNPx) belongs to the family of typical 2-Cys peroxiredoxins. These enzymes function as antioxidants through their peroxidase and peroxynitrite reductase activities. In T. cruzi, as in all trypanosomatids, this enzyme is the final electron acceptor of a unique system for detoxifying hydroperoxides, constituting a relevant target for drug design. We have determined the crystal structure of TcTXPNx in the reduced active state. The structure comprises 10 subunits in the asymmetric unit, associated to form a decamer of toroidal shape obeying 52 (D5) point group symmetry. We have analyzed the structure of TcTXNPx by comparing it with other structures of typical 2-Cys peroxiredoxins in both redox states, and have identified key residues in the structural rearrangement taking place in the enzymatic cycle. This is the first report of the structure of an active peroxiredoxin that has peroxidase and peroxynitrite reductase activity, and it is noteworthy that it is from a human parasite. This knowledge is of interest for further understanding peroxide metabolism in these parasites, and in the design of new trypanosomatidal drugs against Chagas disease. PMID- 15797727 TI - Gramicidin structure and disposition in highly curved membranes. AB - With a view to deciphering aspects of the mechanism of membrane protein crystallization in lipidic mesophases (in meso crystallization), an examination of the structure and disposition of the pore-forming peptide, gramicidin, in the lipidic cubic phase was undertaken. At its simplest, the cubic phase consists of lipid and water in the form of a molecular 'sponge.' The lipid exists as a continuous, highly curved bilayer that divides the aqueous component into two interpenetrating but non-contacting channels. In this study, we show that gramicidin reconstitutes into the lipid bilayer of the cubic phase and that it adopts the channel, or helical dimer, conformation therein. Fluorescence quenching with brominated lipid was used to establish the bilayer location of the peptide. Electronic absorption and emission spectroscopies corroborated this finding. Peptide conformation in the cubic phase membrane was determined by circular dichroism. The identity and microstructure of the mesophases, and their capacity to accommodate gramicidin and other system components (sodium dodecyl sulfate, trifluoroethanol), was established by small-angle X-ray diffraction. Beyond a limiting concentration, gramicidin destabilized the cubic phase in favor of the inverted hexagonal phase. While gramicidin remained bilayer bound as membrane thickness changed, its conformation responded to the degree of bilayer mismatch with the hydrophobic surface of the peptide. These findings support the hypothesis that reconstitution into the lipid bilayer is an integral part of the in meso crystallization process as applied to membrane proteins. They also suggest ways for improving the process of membrane protein crystallogenesis. PMID- 15797728 TI - Trypsin crystallization by membrane-based techniques. AB - To grow protein crystals is not an easy task; moreover, if we need to grow protein crystals with controlled shape, size, and size distribution, depending on their application, the mission becomes even harder. Membrane crystallization has been recognized as an interesting tool for growing protein crystals with enhanced crystallization kinetics, both in static and in forced solution flow configuration, without detrimental effects on crystal quality. In the present work, we have studied the membrane crystallization process of benzamidine inhibited trypsin from bovine pancreas (BPT), with ammonium sulphate (dissolved in Tris-HCl buffer, 0.1 M, pH 8.5), as precipitant agent. We have demonstrated that, by using the membrane crystallization technique, BPT crystals can be obtained in 24-48 h, in static configuration, and in 4-7 days, in a forced solution flow system, depending on the experimental conditions. Furthermore, the kinetics of BPT crystallization have been modulated, to control the morphological characteristics of the crystals produced, by an accurate selection of the operative parameters involved in the process. The active membrane surface and the flow rate of extraction solvent in quiescent configuration, and the solution velocity in forced convection solution experiments, were the parameters investigated. In this respect, membrane crystallization techniques have been assessed as an interesting way for growing proteins, and more specifically enzyme crystals, with high control on the final properties of the crystalline material produced, with potential fundamental implication in the field of structural biology and biotechnology. PMID- 15797729 TI - Trimeric structure of OprN and OprM efflux proteins from Pseudomonas aeruginosa, by 2D electron crystallography. AB - OprM and OprN belong to the outer membrane factor family of multidrug efflux proteins from Pseudomonas aeruginosa, a bacterium responsible of nosocomial infections. We report here the two-dimensional (2D) crystallization of OprN and OprM into lipid bilayers and the determination of their 2D projected structure by cryo-electron crystallography, at 1 and 1.4 nm, respectively. Both proteins present a dense ring of protein density, of approximately 7 nm diameter. An additional thin peripheral ring is resolved in OprN structure. Both proteins are assembled as trimers. The results presented here indicate a high structural homology between OprN (and OprM) and TolC, a multidrug efflux protein from Escherichia coli. PMID- 15797730 TI - Crystal structure of the N-terminal RecA-like domain of a DEAD-box RNA helicase, the Dugesia japonica vasa-like gene B protein. AB - The Dugesia japonica vasa-like gene B (DjVLGB) protein is a DEAD-box RNA helicase of a planarian, which is well known for its strong regenerative capacity. DjVLGB shares sequence similarity to the Drosophila germ-line-specific DEAD-box RNA helicase Vasa, and even higher similarity to its paralogue, mouse PL10. In this study, we solved the crystal structure of the DjVLGB N-terminal RecA-like domain. The overall fold and the structures of the putative ATPase active site of the DjVLGB N-terminal RecA-like domain are similar to those of the previously reported DEAD-box RNA helicase structures. In contrast, the surface structure of the side opposite to the putative ATPase active site is different from those of the other DEAD-box RNA helicases; the characteristic hydrophobic pockets are formed with aromatic and proline residues. These pocket-forming residues are conserved in the PL10-subfamily proteins, but less conserved in the Vasa orthologues and not conserved in the DEAD-box RNA helicases. Therefore, the structural features that we found are characteristic of the PL10-subfamily proteins and might contribute to their biological roles in germ-line development. PMID- 15797731 TI - Automated acquisition of cryo-electron micrographs for single particle reconstruction on an FEI Tecnai electron microscope. AB - AutoEM is a software package developed by Zhang et al. [J. Struct. Biol. 1356, 251] for semi-automated acquisition of cryo-electron micrographs from Tecnai series electron microscopes and is used frequently at the lowest level of automation. We report here on the new progress that we have made based on their preliminary work. A fourth low-dose state is created where the system can pre select all the good holes in a grid square from a single CCD image taken at low magnification, making the system operative at much higher levels of automation. An additional control interface enables the operator to monitor the status of the program and the quality of the data, interact with the program, and direct the execution process according to intermediate results. When data acquisition is in progress, all useful information is automatically saved in certain text files which are easily accessible by a database. More detailed improvements and general advantages are illustrated and discussed. We have started to use the program to perform routine data collection. A number of applications show that the performance of the program is satisfactory and the quality of the micrographs and their power spectra acquired by the program is comparable to those manually collected under the same conditions. PMID- 15797732 TI - Molecular chemistry imaging to reveal structural features of various plant feed tissues. AB - Synchrotron Fourier transform infrared (FTIR) microspectroscopy as a rapid, direct, and non-destructive analytical technique can explore molecular chemical features of the micro-structure of biological samples. However, the application of this synchrotron technology to feed science and feed chemistry is extremely rare. This article reviews that with synchrotron FTIR microspectroscopy, the molecular chemistry of various feed tissues could be imaged. These images revealed spatial intensity and distribution of chemical functional groups in various feeds tissues within cellular dimensions. Such information can be used for plant breeding program for selecting superior variety of plant for targeted feed purposes and for prediction of feed quality and nutritive value. The final purpose of this article shows that Synchrotron FTIR microspectroscopy can be used for biological structure study. PMID- 15797733 TI - Helical crystallization on lipid nanotubes: streptavidin as a model protein. AB - In this study, we use streptavidin (SA) as a model system to study helical protein array formation on lipid nanotubes, an alternative to 2D studies on lipid monolayers. We demonstrate that wild-type and a mutant form of SA form helical arrays on biotinylated lipid nanotubes. 3D maps from helical arrays of wild-type and mutant SA were reconstructed using two different approaches: Fourier-Bessel methods and an iterative single particle algorithm. The maps show that wild-type and mutant streptavidin molecules order differently. The molecular packing arrangements of SA on the surface of the lipid nanotubes differ from previously reported lattice packing of SA on biotinylated monolayers. Helical crystallization on lipid nanotubes presents an alternative platform to explore fundamentals of protein ordering, intermolecular protein interaction and phase behavior. We demonstrate that lipid nanotubes offer a robust and reproducible substrate for forming helical protein arrays which present a means for studying protein structure and structure-function relationships. PMID- 15797735 TI - Cutting artefacts and cutting process in vitreous sections for cryo-electron microscopy. AB - Cryo-electron microscopy of vitreous sections (CEMOVIS) has recently been shown to provide images of biological specimens with unprecedented quality and resolution. Cutting the sections remains however the major difficulty. Here, we examine the parameters influencing the quality of the sections and analyse the resulting artefacts. They are in particular: knife marks, compression, crevasses, and chatter. We propose a model taking into account the interplay between viscous flow and fracture. We confirm that crevasses are formed on only one side of the section, and define conditions by which they can be avoided. Chatter is an effect of irregular compression due to friction of the section of the knife edge and conditions to prevent this are also explored. In absence of crevasses and chatter, the bulk of the section is compressed approximately homogeneously. Within this approximation, it is possible to correct for compression by a simple linear transformation for the bulk of the section. A research program is proposed to test and refine our understanding of the sectioning process. PMID- 15797734 TI - Prepore to pore transition of a cholesterol-dependent cytolysin visualized by electron microscopy. AB - Perfringolysin O (PFO), a soluble toxin secreted by the pathogenic Clostridium perfringens, forms large homo-oligomeric pore complexes comprising up to 50 PFO molecules in cholesterol-containing membranes. In this study, electron microscopy (EM) and single-particle image analysis were used to reconstruct two-dimensional (2D) projection maps from images of oligomeric PFO prepore and pore complexes formed on cholesterol-rich lipid layers. The projection maps are characterized by an outer and an inner ring of density peaks. The outer rings of the prepore and pore complexes are very similar; however, the protein densities that make up the inner ring of the pore complex are more intense and discretely resolved than they are for the prepore complex. The change in inner-ring protein density is consistent with a mechanism in which the monomers within the prepore complex make a transition from a partially disordered state to a more ordered transmembrane beta-barrel in the pore complex. Finally, the orientation of the monomers within the oligomeric complexes was determined by visualization of streptavidin (SA) molecules bound to biotinylated cysteine-substituted residues predicted to face either the inner or outer surface of the oligomeric pore complex. This study provides an unprecedented view of the conversion of the PFO prepore to pore complex. PMID- 15797736 TI - Pathophysiology of hepatic dysfunction and intrahepatic cholestasis in heart failure and after left ventricular assist device support. PMID- 15797738 TI - To induce or not to induce: do patients at greatest risk for fatal rejection benefit from cytolytic induction therapy? AB - BACKGROUND: Induction immunosuppression utilizing lymphocytolytic agents in the early peri-operative period has a number of theoretical and practical advantages and disadvantages. However, the efficacy of cytolytic agents as induction therapy remains unproven. METHODS: To assess the current impact of induction therapy in heart transplantation, we queried a multi-institutional database regarding the frequency of use, type of agent, duration of therapy and outcomes of 6,553 patients transplanted from 1990 to 2001. A study group of 5,897 patients were identified who survived the first 48 hours post-transplant and received either no induction therapy (n = 4,161) or induction with OKT3 or anti-thymocyte preparations (n = 1,736). RESULTS: By multivariate analysis, risk factors for rejection death were identified and then applied to a model of overall mortality. Among patients with a 1-year risk of rejection death at >5%, induction therapy provided a survival advantage, but survival with induction was decreased when the risk of rejection death was <2%. Specific patient sub-sets that received a survival benefit in the current era with induction included younger patients of black race with >/=4 HLA mismatches and long-term (>6 months) support on a ventricular assist device (VAD). CONCLUSIONS: Use and application of induction therapy continues to be controversial in heart transplantation. At present, this approach appears to be beneficial in selected patients who are at high risk for rejection death, but likely detrimental in patients who are at low risk for rejection death. Those with a combination of longer term VAD support, of black ethnicity, and having extensive HLA mismatching are most likely to benefit from cytolytic induction therapy. PMID- 15797737 TI - Na+/H+ exchange inhibition and antioxidants lack additive protective effects after reperfusion injury in the working heterotopic rat heart isograft. AB - BACKGROUND: The utility of combining strategies of myocardial protection was studied in intact rat hearts subjected to 1 hour of ischemia and 40 minutes blood reperfusion. METHODS: Lewis rats (n = 48) were divided into 4 transplant groups. Twenty-four hearts were arrested by coronary perfusion with hypothermic Celsior solution at 60 mm Hg. The aortic valve was punctured to introduce volume into the left ventricle (LV), and the hearts were abdominally isografted. Animals were either given both the antioxidant probucol (300 mg/kg) and the sodium-hydrogen exchange inhibitor cariporide (5 mg/kg) (CP; n = 6), just cariporide (CAR; n = 6), just probucol (PROB; n = 6), or neither drug (CON; n = 6). After 40 minutes of blood reperfusion, transplanted hearts were rearrested. The control recipients' native hearts (native; n = 6) were also arrested. Postmortem LV compliance relations and myocardial water content (MWC) were measured. RESULTS: Grafts protected by probucol were significantly more compliant than controls and significantly less compliant than grafts protected by cariporide alone and with both cariporide and probucol (p = 0.0001, analysis of variance). Compliance relations for CP overlapped those for CAR. All grafts were less compliant than natives. MWC was significantly greater in controls and PROB than in natives. CONCLUSIONS: Pretreatment with cariporide in the setting of ischemia-reperfusion injury provides greater protection against the development of diastolic abnormalities than probucol when Celsior solution is used for both arrest and preservation. In this model, there is no advantage to combining the drugs, supporting the hypothesis that there is an overlapping mechanism of protection. PMID- 15797739 TI - Early mortality after cardiac transplantation: should we do better? AB - BACKGROUND: According to International Society for Heart and Lung Transplantation (ISHLT) data, the 30-day survival after heart transplantation has continually improved from 84% (1979-85) to 91% (1996-2001). This has probably been achieved by better donor/recipient selection, along with improved surgical technique and immunosuppressive therapy. On the other hand, the data concerning the early causes of death after cardiac transplantation is incomplete, because in 25% of cases, an unknown cause is listed. This study investigated the incidence and causes of 30-day mortality (determined by postmortem studies) after cardiac transplantation and assessed the possibility of improvements. METHODS: A retrospective study of all patients who underwent heart transplantation at Papworth Hospital from 1979 to June 2001 (n = 879) and who died within 30 days of surgery was carried out. Postmortem examination data were available for all patients. RESULTS: The mean (standard deviation) recipient and donor ages were 46 (12) and 31 (12) years, respectively. Overall, the 30-day mortality was 8.5% (n = 75), 12.1% for the 1979 to 1985 period and 6.9% for the 1996 to 2001 period. The primary causes of death were graft failure (30.7%), acute rejection (22.7%) (1.3% for the 1996-2001 era), sepsis (18.7%) gastrointestinal problems (bowel infarction and pancreatitis; (9.3%), postoperative bleeding (6.7%), and other (12%). CONCLUSIONS: Our 30-day mortality compares favorably with the data from the ISHLT registry, with great improvement in the early mortality. Acute rejection is no longer a major cause of early mortality. Further reduction may be achieved by a better protection of the donor heart against the effects of brainstem death and ischemic injuries. However, the quest to improve early outcome should not be at the expense of needy patients by being overselective. PMID- 15797740 TI - Angiotensin converting enzyme insertion/deletion polymorphism does not influence postcardiac transplantation hypertension onset or progression. AB - BACKGROUND: The angiotensin converting enzyme insertion deletion polymorphism (ACE I/D) has been associated with much cardiovascular pathology, including posttransplantation hypertension. Hypertension is a significant cause of morbidity and mortality after cardiac transplantation. We investigated the influence of the ACE I/D polymorphism on posttransplantation hypertension. METHODS: A total of 211 heart transplant recipients and 154 corresponding donors were genotyped for the ACE I/D polymorphism by polymerase chain reaction. ACE enzymatic activity was measured by spectrophotometric kinetic analysis. Sitting systolic and diastolic blood pressures were recorded at 3 consecutive visits, and the mean was calculated. Clinical data, including demographics and medication, were collected for all recipients. Results were analyzed by the chi-square test and analysis of variance, taking a p value of <0.05 to be significant. RESULTS: A total of 41.7% of the subjects were hypertensive (diastolic blood pressure >90 mm Hg) at the time of the study, with 79.6% taking at least one antihypertensive agent. We found no difference between the number of antihypertensive agents, cyclosporin dose and level, renal function, or systolic blood pressure for the different recipient or donor genotypes. We also found no significant correlation between ACE enzymatic activity and systolic or diastolic blood pressure. CONCLUSIONS: Our study of 211 recipients and 154 corresponding donors is the largest investigation of this polymorphism in a cardiac transplantation population. We found no apparent relationship between the ACE genotype (of either donor or recipient) and systemic hypertension (absolute measurements and the number or dose of antihypertensive agents used). PMID- 15797741 TI - Diagnostic validity of hepatocyte growth factor as marker for rejection in the follow-up of patients after heart transplantation. AB - BACKGROUND: Hepatocyte growth factor (HGF) is a member of growth factor with a variety of known activities, including angiogenesis promotion and antiapoptotic action. It prevents acute graft-versus-host disease after bone marrow transplantation and prolongs allogenic graft survival in rats. The aim of the study was to investigate the relationship between serum HGF concentration and the grade of acute cellular rejection of heart transplant. METHODS: We studied 68 male heart recipients. All of them received triple-drug immunosuppression: cyclosporine A, prednisone, and azathioprine or mycofenolate mofetil. All patients were without signs of heart failure. Blood samples were taken before elective ambulatory endomyocardial biopsy. Biopsy specimens were graded with the International Society for Heart and Lung Transplantation scale. Acute cellular rejection grade 3A and higher were considered as significant. Measurement of serum HGF was made by enzyme-linked immunosorbent assay. RESULTS: We found a positive relationship between serum HGF levels and grade of cellular rejection. As an indicator for the detection of cell rejection processes, HGF with cutoff 2000 pg/ml seems to be useful. CONCLUSIONS: The results of this study demonstrate that HGF can be useful as an indicator for heart graft cell rejection. PMID- 15797742 TI - Brain natriuretic peptide levels do not correlate with acute cellular rejection in De Novo orthotopic heart transplant recipients. AB - BACKGROUND: The changes in brain natriuretic peptide (BNP) levels after orthotopic heart transplantation have not been previously described. The use of brain natriuretic peptide levels as a surrogate marker for cellular rejection remains controversial, with conflicting data. METHODS: We prospectively evaluated the potential utility of BNP levels in the first 6 months after transplantation and sought correlation with histologic grade of rejection and hemodynamic status. RESULTS: Thirty-five patients and 265 biopsy samples were included in the study. BNP levels did not correlate with histologic grade of rejection. They showed good correlation with central venous pressure and pulmonary capillary wedge pressure. BNP levels were elevated after transplant and showed a steep time-dependent decline. BNP levels correlated with echocardiographically derived indices of diastolic dysfunction. CONCLUSIONS: BNP levels are not a surrogate marker for rejection in the first 2 months after orthotopic heart transplantation and do not obviate the necessity for endomyocardial biopsy. Whether BNP levels have long term prognostic significance is unclear and remains the subject of ongoing prospective study. PMID- 15797743 TI - Successful use in lung transplantation of an immunosuppressive regimen aimed at reducing target blood levels of sirolimus and tacrolimus. AB - BACKGROUND: The primary aim of this study was to assess the safety and efficacy in lung transplantation of an immunosuppressive regimen aimed at achieving sirolimus and tacrolimus concentrations of 6 to 10 microg/ml and 5 to 7 ng/ml, respectively. METHODS: We retrospectively identified 49 lung transplant recipients who were converted to an immunosuppressive regimen consisting of tacrolimus, sirolimus, and prednisone. Data collected included demographic information, laboratory work, episodes of rejection, bronchiolitis obliterans syndrome (BOS) grade, and adverse effects. RESULTS: The most common reason for conversion to a sirolimus and tacrolimus regimen was BOS. The most common adverse effects were increased triglycerides (10%), leukopenia (8%), and skin rash (6%). Four patients (8%) experienced acute allograft rejection during the study period. We followed BOS grade for 1 year in 23 patients. Of these, BOS grade improved in 8, 13 patients remained unchanged, and 2 worsened. Eleven patients (22%) discontinued sirolimus because of adverse events. CONCLUSION: An immunosuppressive regimen consisting of sirolimus and tacrolimus that aims to keep the trough drug concentrations at 6 to 10 microg/ml and 5 to 7 ng/ml, respectively, provides effective lung allograft protection while maintaining an acceptable side-effect profile. The use of this immunosuppressive combination may have a benefit with regard to BOS. PMID- 15797744 TI - Matrix metalloproteinase induction in post-transplant obliterative bronchiolitis. AB - BACKGROUND: Epithelial cell injury, inflammation, fibrosis, and airway obliteration are associated in post-transplant obliterative bronchiolitis. Fibrosis is a consequence of fibroblastic activity and of collagen deposition after disturbances in the balance of protein formation and degradation. Proteolytic enzymes such as the matrix metalloproteinases mediate degradation. To assess matrix metalloproteinases during obliterative bronchiolitis development, we studied porcine, heterotopic bronchial allografts. METHODS: A total of 119 allografts or autografts were harvested serially at 3 to 60 days after transplantation and processed for histology and in situ hybridization for matrix metalloproteinases 2 and 9. Immunocytochemistry for vimentin and alpha-smooth muscle-cell actin was performed with specific antibodies. RESULTS: Implants had initial ischemic injury to airway epithelium and to the bronchial wall. Recovery was rapid in autografts and in immunosuppressed allografts. In matrix metalloproteinase-2 mRNA activity in fibroblasts, correlation with endothelial expression and expression in macrophages occurred during intense fibroproliferation. We observed intense matrix metalloproteinase-9 positivity during onset of inflammation and fibroproliferation in endothelial cells (p < 0.01), fibroblasts (p < 0.05), macrophages (p < 0.05), and lymphocytes (p < 0.05). Matrix metalloproteinase-9 mRNA activity in fibroblasts correlated with that in endothelial and inflammatory cells and also proved predictive of early obliteration. CONCLUSIONS: Matrix metalloproteinase-2, and especially matrix metalloproteinase-9, gene activity was associated with onset of inflammation and fibroblastic proliferation in allografts, predicting early obliteration. Although this may be the case in the model described, its role in human-allograft post transplant obliterative bronchiolitis requires further supportive data. PMID- 15797745 TI - Tumor necrosis factor gene polymorphism and cardiac allograft vasculopathy. AB - BACKGROUND: Cardiac allograft vasculopathy (CAV) limits survival after cardiac transplantation. Tumor necrosis factor-alpha (TNF-alpha) may be a key factor in the development of CAV. Two bi-allelic polymorphisms associated with high TNF alpha production have been identified in the TNF gene locus, TNFA1/2, at position -308 and TNFB1/2 at +252. We hypothesized that recipient TNFA2 and TNFB2 homozygosity is associated with the development of CAV after heart transplantation. METHODS: TNF gene polymorphisms were analyzed by multiplex fluorescent solid-phase mini-sequencing in 70 cardiac transplant recipients. Recipients homozygous for TNFA2 or TNFB2 (Group A, n = 29) were compared with recipients heterozygous or homozygous for TNFA1 and TNFB1 (Group B, n = 41). Coronary arteriography was performed annually or when indicated. Cumulative freedom from CAV and survival was calculated according to the Kaplan-Meier test. RESULTS: Mean follow-up was 3.8 +/- 0.3 years. In Group A, 11 of 29 recipients (38%) developed CAV compared with 9 of 41 (22%) in Group B (p = 0.12). Cumulative freedom from CAV at 3 years was 42% in Group A and 80% in Group B (p = 0.043). In Group A, 11 of 29 recipients (38%) died during follow-up compared with 4 of 41 (10%) in Group B (p = 0.006). Cumulative survival at 3 years was 72% in Group A and 93% in Group B (p = 0.003). CONCLUSIONS: The results suggest that TNFA2 and TNFB2 allele homozygosity is associated with cardiac allograft vasculopathy and mortality in heart transplant recipients. PMID- 15797746 TI - Cyclosporine treatment of high dose and long duration reduces the severity of graft coronary artery disease in rodent cardiac allografts. AB - BACKGROUND: Graft coronary artery disease (GCAD) limits allograft survival after cardiac transplantation. The objective of this study was to correlate GCAD with the level of immunosuppression in an established allogeneic rodent cardiac chronic rejection model to better understand the mechanisms of GCAD in this system. METHODS: Donor PVG hearts were transplanted into the abdomen of ACI rats. Six recipient groups received either 10, 7.5 or 5 mg/kg/day of oral cyclosporine (CsA), for 90 (10 mg/90 d, 7.5 mg/90 d, 5 mg/90 d) or 10 days (10 mg/10 d, 7.5 mg/10 d, 5 mg/10 d; n = 10 all groups), and grafts procured on Day 90. GCAD was assessed by histology for percent luminal narrowing (%LN), percent affected vessels (%AV) and intima/media ratio (I/M ratio). Sections were stained for ED1 positive macrophages and MHC Class II-positive cells. RESULTS: The 10 mg/90 d treatment group showed significantly reduced GCAD compared with the 5mg/10d treatment group (%LN = 4.3 +/- 3.1% vs 39 +/- 11.9%, p < 0.05). The 7.5 mg/90 d group had a reduced %LN and I/M ratio compared with the 5 mg/10 d group (%LN = 8.0 +/- 3.5% vs 39 +/- 11.9%, p < 0.05; I/M ratio = 0.06 +/- 0.02 vs 0.41 +/- 0.14, p < 0.05). There was a trend toward reduction of GCAD with both increasing the dose of CsA as well as the duration of treatment. Continuous treatment with CsA reduced perivascular macrophage and MHC II cell infiltration. Macrophage infiltrates correlated strongly with GCAD (R(2) > 0.90, p < 0.01), and MHC II infiltrates showed a weak correlation, although not statistically significant (R(2) > 0.56, p = NS). CONCLUSIONS: This study further defines the effect of cyclosporine on GCAD in this cardiac transplantation model. In this system, higher dose and longer duration of treatment with CsA markedly reduces macrophage and MHC II infiltration, correlating with diminished GCAD. However, increasing dose and duration of CsA did not completely eliminate the development of GCAD. Non-immunologic factors could contribute to this occurrence. PMID- 15797749 TI - Antiproliferative effects of NKH477, a forskolin derivative, on cytokine profile in rat lung allografts. AB - OBJECTIVE: NKH477 was recently identified as a water-soluble forskolin derivative and was reported to prolong survival of murine cardiac allografts. However, the mechanism of the efficacy is not clear in vivo. The aim of this study was to investigate the immunosuppressive effects of NKH477 on acute lung allograft rejection in the rat model and its mechanism of action in vivo. METHODS: Left lungs were transplanted orthotopically from Brown-Norway donors to Lewis recipients. Recipient rats were untreated or treated daily with different doses of NKH477. Grafts were excised on Day 3 or Day 5 to determine histopathological rejection and expressions of interleukin (IL)-2, IL-4, IL-10, and interferon (IFN)-gamma by enzyme-linked immunosorbent assay. The cytokine expression at Day 3 or Day 5 was also evaluated in recipient spleens by immunohistochemistry. Furthermore, mesenteric lymph node cells from recipients at Day 5 were cultured alone or stimulated with donor antigens for 72 hours to determine cell proliferation by means of thymidine incorporation. RESULTS: NKH477 significantly extended allograft survival time in a dose-dependent manner and reduced histopathological rejection. Treatment with NKH477 inhibited IFN-gamma and IL-10 expression, whereas expression of these cytokines were markedly upregulated in the untreated allografts. Expression of IL-2 and IL-10 also increased in the spleen of untreated allorecipients. NKH477 suppressed expression of both cytokines in the spleen. In addition, lymphocyte proliferation was inhibited in NKH477-treated recipients as compared with untreated recipients. CONCLUSION: These results suggest that NKH477 exerts an antiproliferative effect on lymphocytes in vivo with an altered cytokine profile in rat recipients of lung allografts. PMID- 15797748 TI - Fluorescence imaging microscopy of cellular markers in ischemic vs non-ischemic cardiomyopathy after left ventricular unloading. AB - BACKGROUND: The heart undergoes repair and initiates protective mechanisms via ventricular unloading. We examined the presence of 2 markers in pre-unloaded and post-unloaded human cardiac tissue that are important indicators of cardiac failure, tumor necrosis factor-alpha and inducible nitric oxide synthase. We also measured 2 nuclear transcription factors, NFkappaB50 and NFkappaB65, comparing quantities and localizations to determine if mechanical unloading reduced their presence, as these markers are also thought to be indicators of impending heart failure. Amounts and localizations in patients that had been diagnosed with either ischemic or non-ischemic cardiomyopathy were compared after mechanical unloading with a left ventricular assist device. To establish that unloading had been achieved, levels of atrial natriuretic protein were determined. METHODS: Core biopsies were harvested at assist device implantation and removal. Fluorescence deconvolution microscopy image reconstructions of fluorescence probes were correlated with data obtained by western Blot and electrobility shift assays. RESULTS: Statistically significant differences in localization and amounts of tumor necrosis factor and nitric oxide synthase were seen between pre- and post-assist device samples. Amounts of tumor necrosis factor and nitric oxide synthase in ischemic tissue were increased at the time of assist device removal, but decreased in dilated or idiomyopathic samples. Ventricular unloading resulted in reduced levels of natriuretic protein, with the greatest reduction being seen in ischemic tissue. Both NFkappaB50 and NFkappaB65 increased in ischemic tissue, but only NFkappaB50 in non-ischemic samples. CONCLUSIONS: Changes in localization of the factors and altered levels of cytokine and nitric oxide synthase indicate that the heart switches to a "protective and repair" mode, and mechanical unloading allows this transition to occur. Observed changes were dependent on the etiology of the disease. PMID- 15797750 TI - Short- and long-term outcomes of using pulmonary allograft donors with low Po2. AB - BACKGROUND: The establishment of lung transplantation as a treatment modality for end-stage lung disease has led to an imbalance in the demand and supply for such a procedure. Increasingly marginal donors are being accepted for transplantation. We assessed the short- and long-term outcomes with the use of lung donors with low Po(2). METHODS: All heart-lung and double lung transplantations (n = 362) carried out between 1984 and 2001 were included. Recipients were divided according to the optimized donor Po(2) (on 100% Fio(2)): Po(2) = 30 to 40 kPa = low Po(2) donors (n = 50) and Po(2) >40 kPa = normal Po(2) donors (n = 312). There were no differences in the sex distribution, cytomegalovirus infection status, ischemic time, and intubation durations for the recipients and their respective donors between the 2 groups. The low Po(2) donors were older (38 vs 32 years, p = 0.01) and the allografts were transplanted into younger recipients (33 vs 38 years, p = 0.01). RESULTS: There was a trend toward an increase in the 30 day mortality between the 2 groups (22% vs 13%, odds ratio 1.92, 95% confidence interval 0.91-4.05 p = 0.08). The 1- and 5-year survival rates (standard error) were 66% (7%) and 52% (7%) for the low Po(2) group and 72% (3%) and 44% (3%) for the normal Po(2) group (p = 0.97). Similar infection rates were recorded for the groups. Although rejection rates were similar in the first 3 months, there was a lower rate of rejection in the low Po(2) group thereafter, (hazard ratio, 0.52; p = 0.05). Risk of bronchiolitis obliterans syndrome (BOS) onset was marginally increased in the borderline donors (hazard ratio 1.05, 95% confidence interval 0.68-1.62), although this was not statistically significant. CONCLUSIONS: Donor lung allograft, with optimized Po(2) between 30 and 40 kPa on 100% Fio(2), used for lung transplantation did compromise 30-day mortality, but the difference in mortality did not extend beyond 30 days in our patient group. PMID- 15797747 TI - Cyclosporine mitigates graft coronary artery disease in murine cardiac allografts: description and validation of a novel fully allogeneic model. AB - BACKGROUND: The effect of cyclosporine (CsA) on the development of graft coronary artery disease (GCAD) is controversial. We developed a novel allogeneic mouse model of heart transplantation and investigated the effect of CsA on acute rejection and GCAD. METHODS: Hearts of FVB mice (H-2(q)) were heterotopically transplanted into 60 C57BL/6 mice (H-2(b)). CsA was administered to recipients at 10, 20 or 30 mg/kg/day for 10 or 30 days after transplantation. Untreated recipients as well as isograft recipients served as controls. Viability of the grafts was assessed daily by palpation. Parenchymal rejection was scored in grafts surviving 30 days in the 30-day treatment groups. GCAD was evaluated by the percentage of luminal narrowing, intima/media ratio and percentage of diseased vessels. Blood CsA and creatinine levels were also evaluated. Results were evaluated statistically. RESULTS: All groups except the untreated control group and the allograft groups treated with 10 or 20 mg for 10 days showed significant graft survival (>/=33% survival for 30 days). An inverse correlation was observed between CsA treatment dose, parenchymal rejection score and degree of GCAD in the 30-day treatment groups. However, graft survival in the 20 mg/kg/day group was significantly better than that in the 30-mg/kg/day group. Serum creatine levels showed no nephrotoxicity. CONCLUSIONS: Relatively high-dose CsA mitigated parenchymal rejection and GCAD of the mouse cardiac allografts. In addition, a valuable mouse model mimicking the clinical course of GCAD was achieved with CsA treatment of 20 mg/kg/day for 30 days. PMID- 15797751 TI - Quality of life after lung transplantation: a cross-sectional study. AB - BACKGROUND: The increase in life expectancy has raised the importance of quality of life after lung transplantation, although this issue has not yet been systematically investigated. The aim of this cross-sectional study was to provide an overview of quality of life in lung transplant patients. METHODS: Between April and July 2001, a set of 4 questionnaires were given to all adult, German speaking lung transplant recipients (n = 108; response rate 87% [n = 94]): St. George's Respiratory Questionnaire (SGRQ), a lung-specific health status instrument; the Short Form-36 (SF-36), a general quality of life measure; the Hospital Anxiety and Depression Scale (HADS-D) and a self-reported questionnaire concerning sexual functioning, physical symptoms, adverse effects of immunosuppression and satisfaction with outcome. RESULTS: With regard to the SGRQ, all lung transplant recipients had significantly better scores, which reflected an enhanced quality of life compared with the reference value for patients with obstructive pulmonary disease (SGRQ: p < 0.001 in all sub-scales). The Hospital Anxiety and Depression Scale and the Mental Component Summary Scale of the SF-36 did not differ significantly compared with published normative data in the healthy population. However, the side effects of immunosuppression proved to be a major factor influencing the quality of life of lung transplant recipients (11% influenced strongly and 24% moderately). Seventy-six percent of lung transplant patients were highly satisfied with the transplant outcome and 92% would opt for the procedure again. Among all lung transplant recipients, those with cystic fibrosis were most satisfied (significantly) with their quality of life. CONCLUSIONS: Although lung transplant patients must cope with the side effects of immunosuppression, they report a highly satisfying quality of life with regard to physical and emotional well-being and social and sexual function. PMID- 15797752 TI - Different applications for left ventricular mechanical support with the Impella Recover 100 microaxial blood pump. AB - The "Impella Recover 100" (IR100) is a new intravascular microaxial blood pump for use as short-term mechanical support for cases of acutely reduced left ventricular function. From September 2002 to April 2003, we used the IR100 to support 5 patients: 2 patients were bridged to heart transplant; 2 were being treated for fulminant acute myocarditis; and 1 for post-cardiotomy low-output syndrome. Only 1 patient with myocarditis died of septic shock, 2 had successful heart transplants; and the latter 2 were slowly weaned from the device and, at 3 month follow-up, showed moderate improvement of left ventricular (LV) function. Our initial experience with the IR100 as mechanical support for patients in cardiogenic shock of varying etiology has been positive, yielding good survival in a population of particularly compromised patients. PMID- 15797753 TI - Cardiac transplantation in patients with iron overload cardiomyopathy. AB - A review of the published world experience with heart transplantation for iron overload cardiomyopathy (IOC) between 1967 and 2003 as well as review of unpublished cases from the database of the United Network for Organ Sharing since 1992, identified a total of 16 patients (14 men and 2 women). Mean age was 31 years (range, 14-63 years). IOC etiology was hemochromatosis in 11 patients (69%), thalassemia major in 4 (25%), and Diamond-Blackfan anemia in 1 (6%). The 30-day mortality was 12%. Three patients (19%) died within 1 year of the transplant, all of infectious complications. An additional patient died at 7.14 years (unknown cause). The actuarial Kaplan-Meier 1-, 3-, and 5-year survival rates were 81% for all 3 time intervals. The actuarial 10-year survival was 41%. PMID- 15797754 TI - Bronchial hyperresponsiveness and the bronchiolitis obliterans syndrome after lung transplantation. AB - Because bronchial hyperresponsiveness has been linked to the bronchiolitis obliterans syndrome (BOS), we determined PD(20) methacholine (PD(20(M))), PD(15) hypertonic saline (PD(15(HS))) and their dose-response slopes (DRS(M) and DRS(HS)) in 8 single and 18 double lung transplant recipients within 1 year of lung transplantation and examined the relationship to bronchoalveolar lavage cell profiles and subsequent development of BOS. Twenty-two patients (81%) had a positive methacholine and 6 (25%) a positive hypertonic saline challenge. A positive PD(15(HS)) was associated with an increased risk for BOS at 2 years (odds ratio 12.6, 95% confidence interval 1.3-123.5, p < 0.05), and time to BOS was significantly and negatively related to DRS(HS) (r = -0.5, p < 0.05) - that is, the greater the response, the shorter the time to BOS. Interestingly, DRS(HS) correlated positively with recipient:donor total lung capacity ratio (r = 0.5, p < 0.05), but there was no relationship between either challenge result and airway inflammation. Methacholine hyperresponsiveness is common after lung transplantation but is not prognostic, whereas response to hypertonic saline may reflect recipient:donor size matching and provide useful information regarding the potential for BOS development. PMID- 15797755 TI - Functional heme oxygenase-1 promoter polymorphism in relation to heart failure and cardiac transplantation. AB - We investigated the functional promoter repeat polymorphism (GT)(n) in the oxidative stress-induced enzyme heme oxygenase-1 in relation to heart failure and complications after transplantation. No influence of the polymorphism on the development of heart failure, graft survival, acute rejection, or transplant atherosclerosis was found. PMID- 15797756 TI - Novel use of sildenafil in the treatment of portopulmonary hypertension. AB - Portopulmonary hypertension is a poorly understood and uncommon complication of advanced chronic liver disease. Current therapy is based largely on treatment options proven in idiopathic pulmonary hypertension. The severity of the portopulmonary hypertension should best be attenuated medically before attempting combined liver and lung transplantation to avoid increased peri-operative mortality. This case report describes the successful use of sildenafil to decrease the pulmonary vascular resistance in a patient with hepatitis-C cirrhosis who was preparing for liver transplantation. PMID- 15797757 TI - Reversal of pulmonary hypertension and subsequent repair of atrial septal defect after treatment with continuous intravenous epoprostenol. AB - We report the first case in the world literature of a patient with an atrial septal defect, severe pulmonary hypertension, and equalization of pulmonary and systemic pressures, who underwent successful closure of an ASD following prolonged therapy with the intravenous vasodilator epoprostenol. Judicious use of continuous prostacyclin in apparently inoperable patients with congenital heart disease may be associated with significant reversal of pulmonary hypertension, and conversion to an operable state. PMID- 15797758 TI - Instability of cytokines at body temperature. PMID- 15797759 TI - Cecal perforation as a rare initial manifestation of post-transplant lymphoproliferative disorder after cardiac transplantation. PMID- 15797760 TI - Prescription of exercise to heart transplantation recipients: the role of respiratory rate. PMID- 15797768 TI - Physiological properties of retinal Muller glial cells from the cynomolgus monkey, Macaca fascicularis--a comparison to human Muller cells. AB - Retinae from rabbits and laboratory rodents are often used as 'models' of the human retina, although there are anatomical differences. To test whether monkey eyes provide a better model, a physiological study of Muller glial cells was performed comparing isolated cells and retinal wholemounts from the cynomolgus monkey, Macaca fascicularis and from man. The membrane conductance of Muller cells from both species was dominated by inward and outward K(+) currents. Cells displayed glutamate uptake currents and responded to nucleotides by intracellular Ca(2+) increases. However, there were also species differences, such as a lack of GABA(A) receptors and of Ca(2+)-dependent K(+) currents in monkey cells. Thus, the use of Muller cells from cynomolgus monkeys may be advantageous for investigating a few specific properties; in general, monkey cells are no more similar to human cells than those from standard laboratory animals. PMID- 15797769 TI - "Phase capture" in amblyopia: the influence function for sampled shape. AB - This study was concerned with what stimulus information humans with amblyopia use to judge the shape of simple objects. We used a string of four Gabor patches to define a contour. A fifth, center patch served as the test pattern. The observers' task was to judge the location of the test pattern relative to the contour. The contour was either a straight line, or an arc with positive or negative curvature. We asked whether phase shifts in the inner or outer pairs of patches distributed along the contour influence the perceived shape. That is, we measured the phase shift influence function. Our results, consistent with previous studies, show that amblyopes are imprecise in shape discrimination, showing elevated thresholds for both lines and curves. We found that amblyopes often make much larger perceptual errors (biases) than do normal observers in the absence of phase shifts. These errors tend to be largest for curved shapes and at large separations. In normal observers, shifting the phase of inner patches of the string by 0.25 cycle results in almost complete phase capture (attraction) at the smallest separation (2 lambda), and the capture effect falls off rapidly with separation. A 0.25 cycle shift of the outer pair of patches has a much smaller effect, in the opposite direction (repulsion). While several amblyopic observers showed reduced capture by the phase of the inner patches, to our surprise, several of the amblyopes were sensitive to the phase of the outer patches. We used linear multiple regression to determine the weights of all cues to the task: the carrier phase of the inner patches, carrier phase of the outer patches and the envelope of the outer patches. Compared to normal observers, some amblyopes show a weaker influence of the phase of the inner patches, and a stronger influence of both the phase and envelope of the outer patches. We speculate that this may be a consequence of abnormal "crowding" of the inner patches by the outer ones. PMID- 15797770 TI - Recognizing partially visible objects. AB - In two experiments we measured object recognition performance as a function of delay. In Experiment 1 we presented half of an image of an object, and then the other half after a variable delay. Objects were subdivided into top versus bottom halves, left versus right halves, or vertical strips. In Experiment 2 we separated the low (LSF) and high spatial frequency (HSF) components of an image, and presented one component followed by the other after a variable delay. For both experiments, performance was worse with a 105ms delay between the presentations of the object components than when the two components were presented simultaneously. These results are consistent with predictions made by models that combine information at a relatively early stage in processing. In addition, the results revealed that object recognition performance is significantly better when the LSF sub-image preceded the HSF sub-image than when the HSF sub-image preceded the LSF sub-image, consistent with previous work suggesting that LSF information is processed prior to HSF in object recognition. PMID- 15797771 TI - The nature of synthetic face adaptation. AB - Recent evidence demonstrates that adapting to a face will systematically bias the perception of faces that lie along the same identity trajectory in geometric face space but not faces that lie along different identity trajectories. We explored this configural aftereffect using synthetic face stimuli developed to measure face-specific processing. Adapting to synthetic "anti-faces" resulted in an identity-specific aftereffect that was characterized by a marked decrease in the slope of the psychometric functions. Adaptation transferred across different face sizes, but not different face viewpoints nor faces constructed about a non-mean face. Performance was captured by a model where responses were modulated through a divisive gain control and an additive constant reflecting a shift in the origin of perceived face space. Together, these results suggest that face adaptation reflects activity from mechanisms common to various processing stages along the visual pathway. PMID- 15797772 TI - Parafoveal preview benefit in reading is not cumulative across multiple saccades. AB - Two empirical predictions can be generated from recent parallel processing models of eye movement control in reading concerning the phenomenon of parafoveal preview benefit. These predictions derive from the assumption that lexical activation accumulates across more than one saccade. A large corpus of eye movement data was used to determine whether parafoveal preview benefit of a target word is modulated by (1) its distance from the penultimate fixation, and (2) the amount of time it spends in the 'perceptual span'. There was an unexpected inverse relationship between first fixation duration and distance: fixations were longer the closer the penultimate fixation was to the target word, and a positive relationship between fixation duration and time: fixations were longer the greater the summed durations of the ultimate and penultimate fixations. These findings represent a challenge for computational models of eye movements in reading. PMID- 15797773 TI - What is the signal in noise? AB - Visual perception is limited by both the strength of the neural signals, and by the noise in the visual nervous system; however, little is known about what aspects of the input noise the human visual system is sensitive to, i.e., what is the signal in noise? In order to investigate this question we asked observers to discriminate differences in the strength of one-dimensional white noise. We measured their response consistency and classification images and compared the results with an ideal energy detector. Our results and modelling show that discrimination of noise is limited by the observer's template (i.e., the weighted combination of energy in each stimulus component) plus higher order nonlinearities (systematic noise), and by sources of random internal noise. We found that systematic noise is present only near detection threshold. Surprisingly, we found that the human template is "adaptive"-- its shape depends on the spatial frequency band of the noise-suggesting that sensitivity to spatial noise is not simply determined via passive filtering. PMID- 15797774 TI - Divergence eye movements are dependent on initial stimulus position. AB - Previous studies on the speed and latency of convergence and divergence eye movements have produced varied, sometimes contradictory, results. Four subjects were studied and tracked 4 degrees disparity step changes for convergence and divergence at different initial target positions. Here we report that the dynamics of divergence movements not only differ from convergence movement, but depend on the initial vergence position. Velocities of divergence eye movements in response to targets that were initially near to the subject were approximately twice that of responses to initially distant targets and also exhibited shorter temporal properties. Hence, while convergence responses are fairly similar irrespective of the initial position, divergence dynamic and temporal properties are dependent on the initial stimulus position. It is speculated that the differences observed in divergence may be the result of nonlinear properties of the extraocular muscles or a difference in the underlying neural controller potentially a difference in the magnitude of the fusion initiating component of divergence. PMID- 15797775 TI - Stereoscopic depth aftereffects without retinal position correspondence between adaptation and test stimuli. AB - To clarify whether stereo-slant aftereffects are independent of stimulated retinal position, two experiments compared the magnitude of aftereffects between the following two conditions: when the adaptation and test stimulus fell on (1) the same retinal position, and (2) on different retinal positions separated by 0.5 degrees -20 degrees . In Experiment 1, disc- or ring-shaped surface consisting of random-dots was presented at the central or peripheral visual fields. In Experiment 2, rectangular surface was presented at the upper or lower visual fields. After two minutes inspection of a random-dot stereogram depicting a +/-30 degrees slanted surface, the observer adjusted the slant of the test stimulus to appear fronto-parallel. The results of the experiments showed that significant aftereffects were observed similarly in both conditions. Moreover, the separation nor the stimulus shape scarcely affected the magnitude of the aftereffects. Based on these results we concluded that the depth processing mechanism which operates independently from the stimulated retinal position is responsible for the depth aftereffects we found. PMID- 15797777 TI - The perception of motion transparency: a signal-to-noise limit. AB - A number of studies were conducted to determine how many transparent motion signals observers could simultaneously perceive. It was found that that the limit was two. However, observers required a signal intensity of about 42% in order to perceive a bi-directional transparent stimulus. This signal level was about three times that required to detect a uni-directional motion signal, and higher than was physically possible to achieve in a tri-directional stimulus (in a stimulus in which the different transparent signals are defined only by direction). These results indicate that signal intensity plays an important role in establishing the transparency limit and, as a consequence, implicates the global-motion area (V5/MT) in this process. PMID- 15797776 TI - Attention enhances contrast sensitivity at cued and impairs it at uncued locations. AB - Transient covert attention increases contrast sensitivity at the target location with an informative spatial cue. Here we explored whether an uninformative spatial cue (50% valid with two possible locations) also increases contrast sensitivity and whether contrast sensitivity is altered at the uncued location as compared to the neutral condition. For all four observers, transient covert attention had both a benefit and a cost: it enhanced contrast sensitivity at the cued location and impaired contrast sensitivity at the uncued location at both parafoveal and peripheral positions. These results are consistent with the idea of limited resources, and indicate that transient attention helps control the expenditure of cortical computation. PMID- 15797778 TI - Effects of target and distractor heterogeneity on search for a color target. AB - Experiments were designed to investigate the effects of target and distractor heterogeneity on the threshold for detection of a color target in a search task. In the first two experiments stimuli were chosen so that the target and distractor stimuli varied along one Cardinal axis in color space, while the target differed from distractors along another Cardinal axis. The Cardinal axis signaling the relevant target-distractor difference was consistent from trial to trial within an experiment. When observers searched for a color target among homogeneous distractors but the color of the target and distractors changed from trial to trial there was a small increase in threshold. When the distractors within a display were heterogeneous, and the target color varied from trial to trial there was a larger and more consistent increase in threshold. Varying stimuli along a Cardinal axis other than the Cardinal axis that differentiates target and distractors can impair performance in visual search tasks. Further experiments showed that the presence of heterogeneous distractors had little or no effect on thresholds when location or color cues indicated that these stimuli were irrelevant to the task. Results suggest that the effect of heterogeneity in these experiments is attentional in nature rather than sensory. PMID- 15797779 TI - Inhibition of return is not a foraging facilitator in saccadic search and free viewing. AB - The ability to search and scan the environment effectively is a prerequisite for spatial behavior. A longstanding theory proposes that inhibition of previously attended loci (Inhibition of return; IOR) serves to facilitate exploration by increasing the likelihood to inspect new areas instead of returning to locations that have been inspected before. In this eye movement study we tested whether we could find evidence in favor of this hypothesis. Here we report that IOR does occur during search and free viewing, because we found increased fixation times preceding return saccades (eye movements that return to previously fixated locations). Meanwhile we observed no influence of IOR on the search strategy. Rather than the predicted low number we found many return saccades. Therefore, IOR does not serve as a foraging facilitator in saccadic search and free viewing. We hypothesize that IOR is an intrinsic aspect of shifting attention and gaze direction and furthermore that it is not always advantageous to prevent return saccades. PMID- 15797780 TI - Quantifying object salience by equating distractor effects. AB - It is commonly believed that objects viewed in certain contexts may be more or less salient. Measurements of salience have usually relied on asking observers "How much does this object stand out against the background?". In this study, we measured the salience of objects by assessing the distraction they produce for subjects searching for a different, pre-specified target. Distraction was measured through response times, but changes in response times were not assumed to be a linear measure of distracting potency. The analysis rested on measuring the effects of varying disparities--in size, luminance, and both-between a target object, a key distractor, and other background items. Our results indicate: (1) object salience defined by luminance or size disparity is determined by the ratio between its defining feature value and the corresponding feature value of background items; this finding is congenial to Weber's law for discrimination thresholds. (2) If we define salience as the logarithm of a feature value ratio, then salience increases approximately as fast due to increase in area as due to increase in luminance. (3) The sum of salience arising from object-background disparity in both size and luminance is larger than their vector sum (orthogonal vectors), but smaller than their scalar sum. PMID- 15797782 TI - Spectrophotometric determination of oxiconazole in topical lotion using methyl orange. AB - A spectrophotometric method is described for the determination of oxiconazole in raw material and in topical lotion. This method is based on the reaction of the oxiconazole with methyl orange in buffered aqueous solution of citric acid at pH 2.3. The chromogen, being extractable with dichloromethane, could be measured quantitatively with maximum absorption at 427 nm. The Lambert-Beer law was obeyed in the concentration range of 4.0-14.0 microg ml(-1). A prospective validation of the method showed that the method was linear (r=0.9995), precise (intra-day: CV=1.57% and inter-day: CV=1.50%) and accurate (mean recoveries: 99.69%). The results compared favourably with those of the HPLC method. PMID- 15797781 TI - Analysis of essential elements in Pragya-peya--a herbal drink and its constituents by neutron activation. AB - Ayurvedic herbal formulations are a good source of several nutrient elements essential for metabolic processes. Pragya-peya, a herbal drink and its 12 herbal constituents have been analyzed for 7 minor (Al, Ca, Cl, Mg, Na, K, P) and 15 trace (Ba, Br, Co, Cr, Cs, Fe, Hg, La, Mn, Rb, Sc, Se, Th, V, Zn) elements by instrumental neutron activation analysis (INAA). The method involves thermal neutron irradiation in a reactor followed by counting at several intervals. Also Cd, Cu, Ni and Pb contents were determined by atomic absorption spectrometry (AAS). Elemental data were validated by simultaneously analysing reference materials (RMs) such as mixed Polish herbs (INCT-MPH-2) and peach leaves (SRM 1547). Sample homogeneity was tested by analysing samples from three different batches collected at different intervals. Pragya-peya has been found to be especially rich in several nutrient elements such as Ca, K, V, Fe, Mn, Se and Zn whereas no single constituent is enriched in all the nutrient elements. Concentrations of elements are discussed vis-a-vis their medicinal/therapeutic uses. Several elements such as Na, K, P, Ca, Fe, Co, and Zn seem to be well correlated in 12 constituent herbs. PMID- 15797783 TI - Photo-isomerization of fluvoxamine in aqueous solutions. AB - The hydrolysis and photolysis of fluvoxamine, a selective serotonin reuptake inhibitor, in aqueous buffer solutions (pH 5, 7, and 9), in synthetic humic water, and lake waters were investigated in the dark and in a growth chamber outfitted with fluorescent lamps simulating the UV output of sunlight at 25 degrees C. No significant hydrolytic degradation/isomerization was observed for 30 days in all aqueous solutions. However, fluvoxamine was moderately isomerized to its (Z)-isomer by simulated sunlight. The photo-isomerization occurred in two stages. The photo-isomerization occurred rapidly within the first 7 days and slowly thereafter with a rate constant of 0.12-0.19 day(-1) for the first stage and 0.04-0.05 day(-1) for the second stage. Photosensitized rate constants in synthetic humic water and in lake waters were approximately 6-7 times faster than that in pH 9 buffer with the rate constants of 1.15-1.34 day(-1) in the first stage. The (Z)-isomer of fluvoxamine was the only product detected in all aqueous solutions and was identified using LC-ESI-MS. PMID- 15797784 TI - Microbiological assay for the determination of meropenem in pharmaceutical dosage form. AB - Meropenem is a highly active carbapenem antibiotic used in the treatment of a wide range of serious infections. The present work reports a microbiological assay, applying the cylinder-plate method, for the determination of meropenem in powder for injection. The validation method yielded good results and included linearity, precision, accuracy and specificity. The assay is based on the inhibitory effect of meropenem upon the strain of Micrococcus luteus ATCC 9341 used as the test microorganism. The results of assay were treated statistically by analysis of variance (ANOVA) and were found to be linear (r=0.9999) in the range of 1.5-6.0 microg ml(-1), precise (intra-assay: R.S.D.=0.29; inter-assay: R.S.D.=0.94) and accurate. A preliminary stability study of meropenem was performed to show that the microbiological assay is specific for the determination of meropenem in the presence of its degradation products. The degraded samples were also analysed by the HPLC method. The proposed method allows the quantitation of meropenem in pharmaceutical dosage form and can be used for the drug analysis in routine quality control. PMID- 15797785 TI - Drug-drug interaction between diclofenac, cetirizine and ranitidine. AB - The interactions between diclofenac (1), cetirizine (2) and ranitidine (3) were investigated by thermal analyses and UV, IR and (1)H NMR spectroscopic studies. In aqueous solution interaction occurred only between 1 and 2, yielding a high molecular weight (1:1), water insoluble ionic salt. Weak charge transfer (CT) interaction exists between the doubly charged piperazine moiety in 2, acting as an electron acceptor and (1). This CT interaction originates from the aromatic groups in 1. The CT band observed at approximately 315 nm exhibits very low absorbance as a result of the partial neutralization of the two positive charges present in the ionic salt. The IR bands of the mixture have wave numbers at nu 3323.1, 1695.3, and delta 1321.1-1210 cm(-1) indicating the presence of the NH group and the neutralized carbonyl group of 1, as well as the carboxylic group of 2. The 1,2,3-substitutions in the benzene ring of 1 in the mixture appear at 1161.1 cm(-1). The (1)H NMR of the mixed drugs shows singlet, triplet and multiplet proton signals due to the same effect. PMID- 15797786 TI - Simultaneous analysis of the H1-antihistamine acrivastine and the decongestant pseudoephedrine hydrochloride by high-performance liquid chromatography. AB - High-performance liquid chromatography (HPLC) was used for the simultaneous quantification of the H(1)-antihistamine acrivastine and the decongestant pseudoephedrine hydrochloride. Both compounds were detected at the wavelength of 214 nm. The influence of the mobile phase and the detection wavelength was evaluated and optimized. This method was used to assay various samples from studies of the commercial preparation Semprex-D capsules. The method was found to be accurate, specific, selective, rapid, and versatile for use in routine quality control analyses. PMID- 15797787 TI - Determination of active ingredient within pharmaceutical preparations using flow injection mass spectrometry. AB - Two separate pharmaceutical blends, one containing 2% caffeine by weight, the other containing 2% creatine by weight, and 200 mg caffeine tablets were examined in this study. The purpose of the analyses was to determine the feasibility of using flow injection mass spectrometry for the quantitative analysis of active ingredient within a drug product or other form of pharmaceutical preparation. For more precise and accurate measurements, it was necessary to incorporate an internal reference within the samples. Further, flow injection analysis showed to provide quicker, more facile method development than the application of chromatographic separation. Samples were analyzed over an analyte concentration range of 5.0-15.0 microg/mL. Analyte selectivity was obtained through the observance of the (M+H)(+) ions generated by positive electrospray ionization of each of the analytes (m/z 195 for caffeine and m/z 132 for creatine), and accurate quantitation was achieved by determining the ratio of the analyte response versus the response of the incorporated reference compound. Sample-to sample precision in these measurements was less than 3%, recovery values were shown to be accurate to within +/-3% of the actual values, and both analytical methods proved to be linear over the assay range (R(2)> or =0.999). Due to the excellent selectivity and low detection limits available to mass spectrometric detection, flow injection mass spectrometric analysis could be particularly applicable for analysis of formulations that contain either low doses of active ingredient, active ingredient with low solubility, or active ingredient that does not possess a strong chromophore. Additionally, this type of methodology shows to be conducive for rapid method development. PMID- 15797788 TI - Diclofenac sodium multisource prolonged release tablets--a comparative study on the dissolution profiles. AB - The aim of this work was to compare the dissolution behaviour of six diclofenac sodium prolonged release tablets of different brands obtained from the national market. The formulations contain the same amount of drug substance but different types and/or amount of excipients. The influence of these differences in formulation on the release characteristics of the dosage forms was evaluated on the European Pharmacopoeia apparatus 2 (paddle) employing eight different dissolution media in the pH range 1.2-8. Friability and hardness were tested too according to the European Pharmacopoeia. Dissolution profiles obtained from the studied formulations showed that the release characteristics vary considerably among different manufacturers and that even identical formulations show rather dissimilar release profiles in all the studied media. Use of both SIF without pancreatin and SIF without pancreatin containing 1% (w/v) Tween 20 resulted in strong discrimination among products. A correlation between friability and hardness and in vitro dissolution was evidenced for two formulations having identical excipient composition. PMID- 15797789 TI - LC methods for acyclovir and related impurities determination. AB - Acyclovir, guanine, and impurity A have been baseline separated with isocratic conditions at pH=3.0 and run time under 15 min by employing a SB-CN column from Agilent (150 mm x 4.6mm and 3.5 microm). Moreover, when run time was increased to 40 min six impurities (guanine, impurities A, F, G, Vir 3/4 and N(7)) plus acyclovir were separated in the same conditions. The mobile phase consisted of buffer A/acetonitrile 96:4 (v/v), being buffer A:25 mM H(3)PO(4) (Milli-Q H(2)O) brought to pH 3.0 with KOH. The same column provided separation for all the seven impurities described in pharmacopoeia, including impurity C, which coeluted with acyclovir in the previous conditions with a mobile phase prepared with 25 mM phosphoric acid (pH=1.8)/acetonitrile 96:4 (v/v). The method has been validated following ICH guidelines and it has demonstrated to be reliable for acyclovir and its impurities determination. PMID- 15797790 TI - Analysis of diltiazem and its related substances by HPLC and HPLC/MS. AB - Diltiazem (DTZ) is an optically active calcium channel blocker having a benzodiazepine structure. The drug used in therapy is (+)-cis-diltiazem with configuration (2S,3S). To describe the analytical profile of DTZ different stationary phases (RP-18, RP-8, monolithic support) were tested. The best separation of DTZ from A, B, E and F was obtained using as stationary phase a RP 8 or a monolithic RP-18. The characterization of impurities was carried out using two analytical systems, HPLC and HPLC/MS. PMID- 15797791 TI - Enantioselective analysis of the metabolites of hydroxychloroquine and application to an in vitro metabolic study. AB - A one-step chiral method for the quantification of the enantiomers of two hydroxychloroquine (HCQ) metabolites, desethylchloroquine (DCQ) and desethylhydroxychloroquine (DHCQ) by HPLC is described, in addition to its application to the in vitro study of HCQ metabolism in rat liver microsomes. Liquid-liquid extraction was used to extract the enantiomers from microsome samples and the separation was performed on a Chiralpak AD-RH column protected with an RP-8 guard column using hexane:isopropanol (92:8, v/v) plus 0.1% diethylamine as the mobile phase, at a flow rate of 1.0 mL min(-1). The detection was carried out at 343 nm. The method proved to be linear in the range of 50-5000 ng mL(-1) for DCQ enantiomers and 125-2500ngmL(-1) for DHCQ enantiomers, with a quantification limit of 50 and 125 ng mL(-1), respectively. Precision and accuracy, demonstrated by within-day and between-day assays, were lower than 15%. The metabolic study demonstrated that metabolism is stereoselective for HCQ. The major metabolites formed in the incubation of racemic HCQ were (-)-(R)-DCQ and ( )-(R)-DHCQ with R/S ratios of 2.2 and 3.3, respectively. PMID- 15797792 TI - Simultaneous determination of cadmium and lead in medicinal plants by anodic stripping voltammetry. AB - A simple method for the simultaneous determination of Cd and Pb in medicinal plants by differential pulse anodic stripping voltammetry, using a hanging mercury drop electrode, was developed. The pre-concentration of the metals was performed in 0.8 mol L(-1) HCl at -0.73V for 180s. The sample preparation was carried out by dry-ashing 1.0g of finely pulverized plant samples for 2.5h at 500 degrees C. The determination limit of the method was 0.12 and 0.010 mg kg(-1) for Pb and Cd, respectively. The method was applied to the quantification of cadmium and lead in samples of Hypericum perforatum, Mikania guaco, Mikania glomerata and Peamus boldus. The voltammetric method was shown to be useful for the control of contaminants in medicinal plants. PMID- 15797793 TI - Identification of unknown impurities in simvastatin substance and tablets by liquid chromatography/tandem mass spectrometry. AB - Unknown impurities were detected in simvastatin substance and tablets at a 0.2% level using the liquid chromatography technique with UV (DAD) detection. The impurity structures were elucidated by a direct hyphenation of liquid chromatograph to high-resolution mass spectrometer with electrospray ionisation interface using solutions of formic acid in water and in acetonitrile as the mobile phase. Peak tracking was performed using the column-switching technique. Accurate mass measurements by quadrupole time-of-flight mass spectrometer equipped with lock-spray provided information about elemental composition of intact molecules and fragments of impurities. Measurement accuracy for precursor ions was around 3 ppm and for fragment ions between 4 and 13 ppm. Mass resolving power was around 6500. Deduced molecular formulae for A1, A2 and A3 impurities were C(27)H(44)O(6), C(26)H(43)O(6) and C(26)H(41)O(5), respectively. The structures proposed for all three impurities revealed modifications of simvastatin molecule on the lactone ring. Impurity A1, detected in simvastatin tablets, was identified as ethyl ester, while the impurities A2 and A3, detected in simvastatin substance, were identified as methyl ester and methyl ether of simvastatin. The impurity from tablets was synthesized and its structure confirmed by LC-UV, LC-MS/MS, and NMR techniques. PMID- 15797794 TI - The simultaneous determination of selected flavonol glycosides and aglycones in Ginkgo biloba oral dosage forms by high-performance liquid chromatography electrospray ionisation-mass spectrometry. AB - An accurate, precise and sensitive liquid chromatography-tandem mass spectrometric (LC-MS-MS) method was developed for the determination of two flavonol glycosides, rutin and quercitrin, together with the algycone markers, quercetin, kaempferol and isorhamnetin in several Ginkgo biloba solid oral dosage forms. In addition, a novel quercetin glycoside, not yet reported in Ginkgo extracts, was identified. Liquid chromatography was performed using a minibore high-performance liquid chromatography (HPLC) column (150 mm x 2.0mm i.d.) and a one step gradient of acetonitrile-formic acid (0.3%) at a flow rate of 0.5 ml/min. Baseline separation of the five selected flavonol marker compounds was achieved within 20 min at 45 degrees C. Tandem mass spectrometry was performed using electrospray ionisation (ESI) in the negative ion mode. The marker compounds exhibited linearity over the range of 3-26 microg/ml and intra- and inter-day standard deviations were better than 7% and 16%, respectively. All Ginkgo products investigated were found to contain varying amounts of target analytes. PMID- 15797795 TI - On-line chemiluminescence determination protocatechuic aldehyde and protocatechuic acid in pharmaceutical preparations by capillary electrophoresis. AB - Capillary electrophoresis (CE) with on-line direct chemiluminescence (CL) detection was first used in detecting protocatechuic aldehyde (PAH) and protocatechuic acid (PA) in their pharmaceutical preparations. It was found that the weak CL produced from the reaction of luminol with ferricyanide in an alkaline solution was strongly increased by PAH and PA which was separated by CE. Parameters affecting separation process and CL detection have been examined in detail. Under the optimum conditions, the baseline separation of PAH and PA was obtained within 6 min. The relative standard deviation (R.S.D.) for the analysis of PAH and PA was less than 1.1% for the migration time and 1.6% for the peak height. The detection limits (S/N=3) of PAH and PA were 7.0 x 10(-8)M and 5.0 x10(-8)M, respectively. The proposed method has been satisfactorily applied to the determination of PAH and PA in Salivia miltorrhrza pharmaceutical preparations. PMID- 15797796 TI - A novel incubation direct injection LC/MS/MS technique for in vitro drug metabolism screening studies involving the CYP 2D6 and the CYP 3A4 isozymes. AB - A direct injection LC/MS/MS method involving a novel incubation technique was developed for the inhibition screening of CYP 2D6 and CYP 3A4 isoenzymes using dextromethorphan and midazolam as probe substrates. Both assays were performed using an electrospray ionization source in the positive ion mode. Direct injection was possible by using a short C 18, LC column (2 mm x 20 mm) with large particle diameter packing (10 microm). Analytical characteristics of the direct injection technique were studied by examining matrix effects, which showed suppression of the ESI signal between 0.20 and 0.65 min. The retention times for analytes were adjusted to approximately 0.8 min (k'>3), resulting in no matrix effect. Column lifetime was evaluated and determined to be approximately 160 direct injections of the matrix. The precision and accuracy of the control samples for the quantitation of dextromethorphan was between -0.53 and -12.80, and 3.73 and 6.69% respectively. Unlike conventional incubation techniques, incubations were carried out in an autosampler equipped with a heating accessory. This novel incubation method, which involved no stirring of the incubation mixture, estimated the Cl(int in vitro) for dextromethorphan and midazolam in human liver microsomes to be 1.65+/-0.22 ml/(hmg) and 0.861 ml/(min mg) respectively. The autosampler tray maintained uniform temperature and was sensitive to changes in temperature between 33 and 41 degrees C. High-throughput screening was performed using known inhibitors of the CYP 2D6 isozyme, and the system was evaluated for its ability to differentiate between these inhibitors. The strong inhibitor quinidine resulted in a 25.6% increase in t(1/2), the medium potency inhibitor chlorpromazine resulted in an increase of 6.14% and the weak inhibitor primaquine had no significant effect on half-life. This technique involves no sample preparation, demonstrated run times of 2 min per injection and can be fully automated. The method should therefore prove to be a valuable tool in the drug discovery process. PMID- 15797797 TI - A sensitive method for determination of COL-3, a chemically modified tetracycline, in human plasma using high-performance liquid chromatography and ultraviolet detection. AB - COL-3, 6-deoxy-6-desmethyl-4-desdimethylamino-tetracycline, is a matrix metalloproteinase inhibitor currently in clinical development. A HPLC-UV method to quantitate COL-3 in human plasma was developed. COL-3 was extracted from plasma using solid-phase extraction cartridges. COL-3 is separated on a Waters Symmetry Shield RP8 (3.9 mm x150 mm, 5 microm) column with EDTA (0.001 M) in sodium acetate (0.01 M, pH 3.5)-acetonitrile mobile phase using a gradient profile at a flow rate of 1 ml/min for 22 min. Carryover was eliminated by using an extended needle wash of methanol:acetonitrile:dichloromethane (1:1:1, v/v/v). Detection of COL-3 and the internal standard, chrysin, was observed at 350 nm. COL-3 and chrysin elute at 8.9 and 9.9 min, respectively. The lower limit of quantitation in human plasma of COL-3 was 75 ng/ml, linearity was observed from 75 to 10,000 ng/ml. A 30,000 ng/ml sample that was diluted 1:50 with plasma was accurately quantitated. This method is rapid, widely applicable, and suitable for quantifying COL-3 in patient samples enabling further clinical pharmacology characterization of COL-3. PMID- 15797798 TI - In vitro plasma protein binding determination of flunarizine using equilibrium dialysis and liquid chromatography-tandem mass spectrometry. AB - A highly sensitive method was developed and validated for determining the free fraction of flunarizine in human plasma. Equilibrium dialysis was used for the separation of free (unbound) drug and liquid chromatography/tandem mass spectrometry (LC-MS/MS) was used for quantitation. Post-dialysis plasma or buffer samples of 0.2 mL were extracted using a liquid-liquid extraction procedure and analyzed using a high performance liquid chromatography electrospray tandem mass spectrometer system. The compounds were eluted isocratically on a Supelco Supelcosil ABZ+Plus column, ionized using a positive ion atmospheric pressure electrospray ionization source, and analyzed using multiple reaction monitoring. The ion transitions monitored were m/z 405-->203 for flunarizine and m/z 409- >207 for flunarizine-d4 (internal standard, IS). The chromatographic run time was 3.5 min per injection, with retention times of 2.1 min for both flunarizine and IS. The calibration curve for flunarizine was linear over the concentration range of 0.25-2000 ng/mL (r(2)>0.9989) in the combined matrix of human plasma and isotonic sodium phosphate buffer (1:1, v/v) with the lower limit of quantitation of 0.25 ng/mL. The inter-assay coefficient of variability (CV) for the quality control samples was less than 13.5%, and the inter-assay percent nominal was greater than 98.2%. In vitro protein binding of flunarizine was determined at concentrations of 5, 10 and 100 microg/mL using the validated method. Flunarizine was extensively bound to plasma protein with a 0.083+/-0.005% overall percent free drug in plasma and a CV value less than 7.8%. This validated method will be used for the ex vivo assessment of flunarizine protein binding in human plasma from a drug-drug interaction clinical study. PMID- 15797799 TI - High-performance liquid chromatographic determination of enalapril in human plasma by enzyme kinetic analytical method. AB - A high-performance liquid chromatographic method for indirect determination of enalapril in human plasma, was developed and validated. An exogenous angiotensin converting enzyme after drug inhibition was determined by reacting with hippuryl histidyl-leucine to produce hippuric acid (HA) which was inversely proportional to the amount of enalaprilat in plasma. The HPLC was carried out on a Lichrosphere 60RP-select B, C18, 5 microm (125 mm x 4.0 mm i.d.) column at flow rate of 1.0 ml/min. The analysis time per injection was within 6.5 min. The lowest concentration of enalaprilat to be quantitated was 3.0 ng/ml with the acceptable accuracy and precision. This successfully developed method was practically and accurately used for pharmacokinetics and bioequivalent study of enalapril. PMID- 15797800 TI - Generation and destruction of unstable reagent in flow injection system: determination of acetylcysteine in pharmaceutical formulations using bromine as reagent. AB - A flow injection spectrophotometric procedure was developed for determination of acetylcysteine in sachets and liquid formulations. The determination of this drug was carried out by reacting it with bromine chemically generated in flow injection system monitored continuously at 400 nm. Acetylcysteine reacts with bromine causing a decrease in the absorbance that is proportional to the analyte concentration. The bromine in excess was destroyed on-line by an ascorbic acid solution before the discard. The calibration curve for acetylcysteine determination was linear in the concentration range from 1.6 x 10(-4) to 1.6 x 10(-3) mol/l with a detection limit of 8.0 x 10(-5) mol/l. The relative standard deviation (R.S.D.) was lesser than 1.2% for a solution containing 5.3 x 10(-4) mol/l acetylcysteine (n=10), and 60 determinations per hour were obtained. PMID- 15797801 TI - Simultaneous determination of ranitidine and metronidazole in human plasma using high performance liquid chromatography with diode array detection. AB - The development and validation of a simple method for the simultaneous determination of ranitidine and metronidazole in human plasma is described. Plasma samples (250 microL) were deproteinized by precipitation with 60% perchloric acid, centrifuged and the supernatant directly injected into the HPLC. Separation was achieved in isocratic mode with a Shimpak C(18) column and a mobile phase consisting of 10mM potassium dihydrogen phosphate pH 3.5:acetonitrile (90:10, v/v) with UV detection at 315 nm. The method showed good selectivity and sensitivity. Good and consistent recovery for metronidazole and ranitidine was obtained: 96.22+/-3.52 and 95.00+/-4.50% for ranitidine (25-1000 ng/mL) and metronidazole (60-10,000 ng/mL), respectively (n=3). With this one step sample preparation method, both ranitidine and metronidazole could be quantified simultaneously in human plasma with good precision (R.S.D.<15%) and accuracy (bias values below 15%). The limit of quantification for ranitidine and metronidazole were 20 and 40 ng/mL plasma, respectively. PMID- 15797803 TI - Simultaneous estimation of pharmacokinetic properties in mice of three anti tubercular ethambutol analogs obtained from combinatorial lead optimization. AB - Integrating combinatorial lead optimization of [1,2]-diamine core structure based on ethambutol with high-throughput screening has led us to focus on three promising analogs (SQ37, SQ59 and SQ109) as potential anti-tubercular drug candidates from thousands of synthesized diamine analogs for further characterization of their biopharmaceutical and pharmacokinetic properties by using liquid chromatography/tandem mass spectrometry (LC/MS/MS) and cassette dosing for pharmacokinetic screening. Simultaneous separation of the three analogs was achieved on reversed phase HPLC using a gradient mobile phase composed of MeOH/CH(3)COONH(4) (5mM)/trifluoroacetic acid: 80/20/0.1 (v/v/v). After extraction with acetonitrile from biomatrices, samples were analyzed on the LC/MS/MS system in the positive mode using an electrospray ion source. The retention time for the analogs ranged from 3.70 to 4.48 min. Incubation of SQ37 with plasma at 37 degrees C for 6h resulted in its degradation in human and rat plasma (20-35%), but no significant degradation was observed in mouse and dog plasma. SQ59 was relatively stable in the plasma of the four species. SQ109 was degraded in human and dog plasma (30-40%), but stable in mouse and rat plasma during the 6h incubation. A rapid multiple pharmacokinetic screening was taken by cassette dosing of the three analogs to mice and simultaneous analysis of their plasma concentrations. The analogs showed large Vd(ss) ranging from 11,300 (SQ37), 12,800 (SQ109) to 63,900 ml/kg (SQ59). The clearance ranged from 3240 (SQ109), 3530 (SQ37) and 8043 ml/kg/h (SQ59). The elimination t(1/2) ranged from 4.4 to 21.1h dependent on the routes. The oral bioavailability was 5.1 (SQ59), 20.1 (SQ37) and 7.8% (SQ109), respectively. Both SQ37 and SQ109 possess good pharmacokinetic properties. PMID- 15797802 TI - Amperometric biosensor based on horseradish peroxidase for biogenic amine determinations in biological samples. AB - An amperometric biosensor for total biogenic amine determinations, using a carbon paste electrode modified with horseradish peroxidase (HRP) enzyme is described. The HRP immobilization on graphite was made using bovine serum albumin, carbodiimide and glutaraldehyde. The biosensor response was optimized using serotonin and it presented the best performance in 0.1 mol l(-1) phosphate buffer (pH=7.0) containing 10 micromol l(-1) of hydrogen peroxide. Under optimized operational conditions at -50 mV versus SCE, a linear response range from 40 to 470 ng ml(-1) was obtained. The detection limit was 17 ng ml(-1) and the response time was 0.5s. The proposed sensor presented a stable response during 4h under continuous monitoring. The difference of the response between six sensor preparations was <2%. The sensor was applied in the determination of total biogenic amines (neurotransmitters) in rat blood samples with success, obtaining a recovery average of 102%. PMID- 15797804 TI - Monoethylglycinexylidide and lidocaine determination in porcine microsomal preparations. AB - A simple, accurate, and sensitive HPLC analysis of monoethylglycinexylidide (MEGX) and lidocaine in porcine microsome samples is described. Lidocaine and MEGX were measured by direct injection after the addition of the internal standard. Chromatography was performed on a muBondapak C(18) column using an isocratic mobile phase of 0.03 M potassium dihydrogen phosphate:acetonitrile (87:13), pH 5.9. UV absorbance was measured at 205 nm. The procedure produced linear curves for the concentration range 50-1000 ng/mL with a limit of detection of 10 ng/mL. Recoveries for both compounds were greater than 90%. This assay produced accurate and repeatable results. PMID- 15797805 TI - Simultaneous determination of gallic acid, albiflorin, paeoniflorin, ferulic acid and benzoic acid in Si-Wu decoction by high-performance liquid chromatography DAD method. AB - A high-performance liquid chromatographic method was applied to the determination of gallic acid, albiflorin, paeoniflorin, ferulic acid and benzoic acid in Si-Wu decoction and other 13 combinations of the formula. These five compounds were analyzed simultaneously with a Zorbox SB C-18 column by gradient elution using 0.01% (v/v) phosphoric acid-acetonitrile as the mobile phase. The flow rate was 1 ml min(-1), and detection was set at 230 nm. The recovery of the method was in the range of 94.8-103.1%, and all the compounds showed good linearity (r>0.9995) in a relatively wide concentration range. The result indicated that the content of these five compounds changed after decocting process. The contents of paeoniflorin, albiflorin, ferulic acid and gallic acid increased and that of benzoic acid decreased significantly. PMID- 15797806 TI - Application of non-aqueous micellar electrokinetic chromatography to the analysis of active components in radix Salviae miltiorrhizae and its medicinal preparations. AB - A simple, economical and effective non-aqueous micellar electrokinetic chromatography (NAMEKC) method was developed for simultaneous assay of three bioactive components (cryptotanshinone, tanshinone IIA and tanshinone I) in radix Salviae miltiorrhizae and its medicinal preparations for the first time. After optimization of separation conditions, a buffer of 140 mmol l(-1) sodium cholate (SC) in methanol was selected for the separation of the three tanshinones, but baseline separation of tanshinone I and tanshinone IIA in practical samples was not achieved. Therefore, second-order derivative electropherograms were applied for resolving overlapping peaks. Regression equations revealed good linear relationships (correlation coefficients 0.995-0.999) between peak heights in second-order derivative electropherograms and concentrations of the three analytes. The recoveries of three constituents ranged from 91.3 to 105.7%. The results indicated that baseline separation of the analytes was hard to be achieved in practical samples sometimes and second-order derivative electropherograms was applicable for the resolving and analysis of overlapping peaks. PMID- 15797807 TI - Comparison between HPLC and HPTLC-densitometry for the determination of harpagoside from Harpagophytum procumbens CO(2)-extracts. AB - Carbon dioxide (CO(2)) extracts of the secondary roots of Harpagophytum procumbens were quantified by high performance liquid chromatography (HPLC) and high performance thin layer chromatography (HPTLC). An isocratic HPLC method was used for the quantification of the iridoid glucoside harpagoside at 278 nm. A HPTLC assay was developed for the determination of harpagoside after coloration at 509 nm. The diode array detection of both analytical assays were used to examine the purity of harpagoside peaks and compared with the standards, respectively. The assays provide good accuracy, reproducibility and selectivity for the quantitative analysis of harpagoside. The harpagoside contents of 15 different CO(2)-extracts were compared by HPLC and HPTLC-densitometry. The quantitative results of both analytical methods did not show any statistical significance between each other, although a trend to slightly lower mean values could be found for the HPTLC method. PMID- 15797808 TI - A multicommuted flow system for the determination of dextrose in parenteral and hemodialysis concentrate solutions. AB - A new method is presented for the automation of the determination of dextrose in parenteral and hemodialysis solutions. The method is based on multicommutation flow analysis (MCFA) and exploits enzymatic reactions providing a colored derivative that is detected spectrophotometrically (Trinder's method). The reagent, comprising glucose oxidase, peroxidase, 4-hydroxybenzoate and a buffer was obtained from a commercial kit for the determination of glycemia. The flow system used three 3-way solenoid valves operating under computer control. The necessary software was developed for the purpose and compiled in QuickBASIC 4.0 The influence of some operating variables (segment size, number of segments and reactor length) was studied. Calibration curves in the range 0-1g/L presented a slight curvature and were fitted with a second-degree polynomial (h= 0.0632C(2)+0.6039C+0.166, r(2)=0.9973, h being the peak-height (absorbance) and C the concentration in g/L). The method was validated by analyzing artificial samples presenting accurately known concentrations of dextrose, and comparing the results with the known value and with value obtained by polarimetry. Recoveries were in the range 96.6-100.2%, and the difference with the polarimetric analysis was in the range 0.1-3.3%. Precision (R.S.D.,%) was better than 2.4%. Sampling frequency of the system was 90 samples/h, with a reagent consumption of 0.14 mL per sample. PMID- 15797809 TI - Exploiting calnexin expression on phagosomes to isolate Leishmania parasitophorous vacuoles. AB - We have developed a simple scheme for the isolation of parasitophorous vacuoles (PVs) that harbor Leishmania parasites. This scheme exploits the observation that PVs display endoplasmic reticulum molecules, including the transmembrane protein calnexin. The presence of calnexin at the surface of the PVs distinguishes them from late endosomal vesicles of comparable density. As a result, PVs can be isolated by calnexin affinity selection from an enriched PV fraction obtained by sucrose density fractionation. PMID- 15797810 TI - Immunopathogenesis of infection with the visceralizing Leishmania species. AB - Human leishmaniasis is a spectral disease that includes asymptomatic self resolving infection, localized skin lesions, and progressive visceral leishmaniasis. With some overlap, visceral and cutaneous leishmaniasis are usually caused by different species of Leishmania. This review focuses on host responses to infection with the species that cause visceral leishmaniasis, as they contrast with species causing localized cutaneous leishmaniasis. Data from experimental models document significant differences between host responses to organisms causing these diverse syndromes. The visceralizing Leishmania spp. cause localized organ-specific immune responses that are important determinants of disease outcome. Both the Leishmania species causing cutaneous and those causing visceral leishmaniasis require a Type 1 immune response to undergo cure in mouse models. However, during progressive murine infection with the visceralizing Leishmania sp., the Type 1 response is suppressed at least in part by TGF-beta and IL-10 without type 2 cytokine production. This contrasts with the cutaneous species L. major, in which a Type 2 response suppresses type 1 cytokines and leads to murine disease progression. Population and family studies are beginning to elucidate human genetic determinants predisposing to different outcomes of Leishmania infection. These studies should eventually result in a better understanding of the immunopathogenesis and the spectrum of human leishmaniasis. PMID- 15797812 TI - A pore-forming toxin produced by Aeromonas sobria activates Ca2+ dependent Cl- secretion. AB - Bacteria produce many types of hemolysin that induce diarrhea by mechanisms that are not completely understood. Aeromonas sobria hemolysin (ASH) is a major virulence factor produced by A. sobria, a human pathogen that causes diarrhea. Since epithelial cells in the intestine are the primary targets of hemolysin, we investigated the effects of ASH on ion transport in human colonic epithelial (Caco-2) cells. ASH increased short-circuit currents (Isc) in a dose-dependent manner, and it also activated a 125I efflux from Caco-2 cells. ASH-induced Isc increases and 125I efflux activations were both suppressed by low Ca2+ levels in the extracellular solution or by pretreatment with the Ca2+ chlelator BAPTA-AM. Intracellular Ca2+ levels were increased by ASH in a biphasic fashion characterized by a rapid sharp increase (peak 1) followed by a sustained low plateau (peak 2). ASH-induced peak 1 was inhibited by pretreatment with pertussis toxin, indicating that Ca2+ was mobilized from intracellular stores, and peak 2 was induced by an influx of extracellular Ca2+. Peak 2 but not peak 1 was related to Cl- secretion. These results indicate that ASH activates Ca2+-dependent Cl- secretion. PMID- 15797811 TI - Mechanisms underlying Mannheimia haemolytica leukotoxin-induced oncosis and apoptosis of bovine alveolar macrophages. AB - Mannheimia (Pasteurella) haemolytica leukotoxin (LktA) binds to the bovine beta2 integrins (such as LFA-1-CD11a/CD18) and leads to subsequent cellular effects in a dose dependent manner. The objectives of this study were to delineate the mechanisms that underlie LktA-induced oncosis and apoptosis and to examine the role of LktA/LFA-1 interaction in these events. The results demonstrate that LktA induced oncosis proceeds through a LFA-1 and caspase-1 dependent pathway referred to as 'pyrotosis', as well as through a LFA-1- and caspase-1-independent pathway. LktA-induced apoptosis in alveolar macrophages involves activation of caspase-3 and engages the extrinsic and intrinsic pathways of apoptosis, with the extrinsic pathway being dependent on LFA-1 signaling and TNFalpha. PMID- 15797813 TI - SlyA regulates the collagenase-mediated cytopathic phenotype in multiresistant Salmonella. AB - Salmonella enterica serotype Typhimurium phagetype DT104 (DT104) is a foodborne pathogen with a multiresistant phenotype conferred by a genomic-based integron structure designated as SGI1. Recently, a novel cytopathic phenotype was ascribed to several isolates of DT104 recovered from veal calves. This phenotype is dependent upon clg, a gene encoding a collagenase in Salmonella. Using a novel transposon system and an RT-PCR assay for detection of clg expression, we identified SlyA as a regulator of the collagenase-mediated phenotype. The function of SlyA, in regards to clg expression, is to repress the synthesis of Clg. Derepression ensued in the absence of SlyA or in the presence of a truncated version of SlyA with the latter being relevant for maintenance of another virulence aspect mediated by SlyA, i.e. survival within macrophages. The SlyA mediated effect on clg expression was restricted to DT104 and other Salmonella phagetypes and serotypes possessing SGI1 thus suggesting co-regulation by an SGI1 specific component. PMID- 15797814 TI - Quantification of ALS1 gene expression in Candida albicans biofilms by RT-PCR using hybridisation probes on the LightCycler. AB - The fungal pathogen Candida albicans has the ability to grow as a biofilm on synthetic materials. This presents a significant problem in the clinical situation when the organism grows as a biofilm on medical devices resulting in infections which are resistant to antifungal agents. Determining the extent to which certain genes are involved in biofilm formation is an important aspect for the development of strategies to control pathogenic biofilms. ALS1 is a member of the ALS (agglutinin-like sequence) family, the protein products of which are implicated in attachment to endothelial cells and biofilm formation. The expression of ALS1 in biofilms grown on silicone elastomer, a material used in the manufacture of medical devices, and planktonically grown cells was investigated using a novel real-time quantitative reverse transcriptase PCR (q-RT PCR) on the LightCycler. This study demonstrates quantitatively that ALS1 is clearly up-regulated during biofilm growth. The real-time q-RT PCR assay described here has the potential to be used as an indicator of biofilm formation on medical devices. PMID- 15797815 TI - Developing the use of mismatch binding proteins for discovering rare somatic mutations. AB - A method for detecting rare, unknown, somatic mutations could allow presymptomatic cancer screening from human fluids. Immobilized mismatch binding protein can bind DNA heteroduplexes while allowing homoduplexes to be washed away, thus enriching for rare mutations. We examined the potential use, for mutation enrichment, of a fusion protein of maltose binding protein and the mismatch binding protein TaqMutS (MBP-MutS). Unlabeled and fluorescent-labeled oligonucleotides, either perfectly complementary or with single nucleotide mismatches or deletions, were combined to form homo- or heteroduplexes that were then mixed at low ratios of hetero- to homoduplexes and enriched for heteroduplexes. Enrichment was observed using a capillary DNA sequencer. A single base deletion oligonucleotide was enriched by a factor of 29, and a mismatch oligonucleotide was enriched by a factor of 2. N-1 oligonucleotide synthesis fragments were enriched more than were mismatches, suggesting that these deletion fragments may compete for MutS and impede enrichment of mismatches. Purification of oligonucleotides by high pressure liquid chromatography or polyacrylamide gel electrophoresis failed to remove n-1 fragments, thus overcoming this obstacle to enrichment of mismatch mutations may require alternative strategies, such as developing new purification methods or avoiding the use of synthetic oligonucleotides before enrichment. PMID- 15797816 TI - Detection of lymphatic Wuchereria bancrofti in carriers and long-term storage blood samples using semi-nested PCR. AB - The semi-nested PCR was conducted for detection of Wuchereria bancrofti in patients' blood. The primers were designed based on the repetitive DNA sequences of the parasite. The results demonstrated that the semi-nested PCR could detect as little as 0.001 fg of parasite DNA. In addition, the primers showed no PCR amplification from human and other hemoparasites such as Brugia malayi, Plasmodium falciparum and P. vivax DNAs. This technique was used for detection of 18 W. bancrofti infected blood samples with a long-term storage, the data revealed that all samples were positive. The results obtained from this study clearly indicated that the semi-nested PCR is specific, sensitive, and suitable for detection of the disease carriers. PMID- 15797817 TI - Citrus huanglongbing in Sao Paulo State, Brazil: PCR detection of the 'Candidatus' Liberibacter species associated with the disease. AB - Symptoms of huanglongbing (HLB), one of the most serious diseases of citrus in Asia and Africa, have been noticed in March 2004 in the Araraquara region of Sao Paulo State, Brazil. HLB has not been reported previously from America. The causal HLB bacteria, Candidatus Liberibacter africanus in Africa and Candidatus Liberibacter asiaticus in Asia, can be detected in symptomatic citrus leaves by PCR amplification of their 16S rDNA with previously described primers. When this technique was applied to 43 symptomatic leaf samples from the Araraquara region, all PCR reactions were negative. This suggested that a new pathogen, not detected by the above primers, could be involved in HLB in the State of Sao Paulo. Indeed, by using universal primers for amplification of bacterial 16S rDNA, a new liberibacter species, Candidatus Liberibacter americanus, has recently been identified. Specific primers for PCR amplification of the 16S rDNA of Ca. L. americanus have been selected. Using these primers, the new liberibacter could be detected in 214 symptomatic leaf samples tested. The leaves of two additional samples were infected with Candidatus Liberibacter asiaticus, and two further samples contained both Ca. L. americanus and Ca. L. asiaticus. The samples came from 47 farms in 35 municipalities. The psyllid vector of Ca. L. asiaticus, Diaphorina citri, is established in South, Central, and North America (Florida and Texas). Ca. L. americanus could be detected by PCR in several batches of D. citri psyllids collected on symptomatic sweet orange trees infected with Ca. L. americanus, strongly suggesting that D. citri is the vector of Ca. L. americanus. The results reported here confirm the presence of HLB in the State of Sao Paulo. Ca. L. americanus is the most widely distributed pathogen. PMID- 15797818 TI - Similar sequence-free amplification of human glyceraldehyde-3-phosphate dehydrogenase for real time RT-PCR applications. AB - One of the major applications of real time polymerase chain reaction (PCR) is relative quantification, where the expression of a target gene is determined as a ratio to a stably expressed reference gene, the so-called housekeeping gene. Glyceraldehyde-3-phosphate dehydrogenase (GAPD) is a glycolytic enzyme, which is active in all mammalian tissues and is frequently used as housekeeping gene in expression studies. The functional locus maps to human chromosome 12p13, but several GAPD-related sequences, including processed pseudogenes, GenBank homologous sequences and computationally predicted sequences are present along the human genome. Due to the high level of GAPD-related sequences it is very important to avoid genomic DNA amplification when GAPD is used as endogenous control in mRNA quantification. We have outlined a GAPD couple of primers that avoid any genomic DNA amplification for real time reverse transcription PCR applications by SYBR-Green Dye. These new designed primers are an useful and chip alternative to probe technologies, and can carry out specific and reproducible data in mRNA expression studies. PMID- 15797819 TI - PCR identification of Campylobacter coli and Campylobacter jejuni by partial sequencing of virulence genes. AB - The objective of this study was to utilize a multiplex PCR assay for concurrent detection of Campylobacter spp. and C. coli or C. jejuni, using probes derived from genes cadF and ceuE and an undefined virulence gene. A total of 97 Campylobacter strains, isolated from turkey litter (n=74), chicken livers (n=15) and clinical (n=8) samples, were speciated using the PCR-based assay. PCR amplification of the isolates identified a 400-bp cadF gene, conserved in Campylobacter species, an 894-bp ceuE gene, specific for C. coli, and a 160-bp oxidoreductase gene, specific for C. jejuni. The approximately 35 kDa cadF adhesion proteins allow Campylobacter to bind to the intestinal epithelial cells and the 37 kDa ceuE lipoproteins are involved in siderophore transport. Sequencing of the 160-bp undefined gene yielded a 67% protein identical match with a gene encoding an oxidoreductase subunit in C. jejuni. The specificity of the assay was validated on 36 non-Campylobacter strains (11 Gram-positive and 25 Gram-negative bacteria). The PCR assay identified 59% of turkey and 47% of chicken isolates as C. jejuni, and 41% of turkey and 53% of chicken isolates as C. coli. All human isolates were identified as C. jejuni. The specificity of this assay to detect C. coli or C. jejuni was 97%. PMID- 15797820 TI - A DNA microarray for identification of virulence and antimicrobial resistance genes in Salmonella serovars and Escherichia coli. AB - Characterization of antimicrobial resistance and virulence gene profiles provides important information on the potential pathogenicity of bacteria. This information can be used to facilitate prompt and effective treatment of bacterial infections. We developed and tested a PCR-based microarray platform for detecting virulence and antimicrobial resistance genes in Salmonella serovars and Escherichia coli. Twelve Salmonella and seven E. coli isolates were screened for the presence of 25 virulence and 23 antimicrobial resistance genes. All S. Typhimurium DT104 isolates harbored virulence plasmids. E. coli O157:H7 isolates possessed virulence genes typical of enterohemorrhagic E. coli (EHEC), whereas E. coli O126 isolates contained virulence genes characteristic of enteropathogenic E. coli (EPEC) and E. coli O111, O78 and O147 isolates had virulence genes characteristic of enterotoxigenic E. coli (ETEC). Correlation between antimicrobial resistance phenotype and genotype was observed for each isolate. The aadA, tetA, and sulI genes were most commonly detected in bacteria resistant to streptomycin, tetracycline and sulfonamide, respectively. All isolates exhibiting resistance to third generation cephalosporins harbored the bla(CMY-2) and bla(TEM-1) genes. Microarray analysis is an effective method to rapidly screen Salmonella and E. coli for multiple antimicrobial resistance and virulence genes. PMID- 15797821 TI - Application of SSB-like protein from Thermus aquaticus in multiplex PCR of human Y-STR markers identification. AB - Here, we demonstrate the application of SSB-like protein from Thermus aquaticus (TaqSSB) in multiplex PCR of human Y-STR markers identification. The use of thermostable TaqSSB prevents or reduces primer dimmer formation, one of the particular problems known to cause inhibition of primer hybridization to the template and reduction of a number of primers available for annealing in PCR reaction. PMID- 15797822 TI - Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP. AB - The membrane-protein 81 gene (mb-mp81) of Mycoplasma bovis was cloned, sequenced and compared to membrane-protein 81 gene (ma-mp81) of Mycoplasma agalactiae. After alignment of both sequences, specific primers pairs were designed from variable or unchanging nucleotide segments. In this study, we describe the development and optimization of a multiplex-PCR (MPCR) for the rapid detection of M. agalactiae and M. bovis strains. In addition, a simple and rapid PCR restriction fragment length polymorphism (RFLP) assay, using the restriction enzymes AluI, DraI, RsaI and XbaI, is described to distinguish between both species. The results suggest that MPCR and PCR-RFLP assays could be used as an alternative method in routine diagnosis for rapid and specific simultaneous detection of M. agalactiae and M. bovis. PMID- 15797823 TI - Rapid PCR detection of enterohemorrhagic Escherichia coli (EHEC) in bovine food products and feces. AB - Although Escherichia coli (E. coli) O157:H7 is a major cause of foodborne illness, other types of E. coli can also cause illness. E. coli that possess the eae gene for attachment and effacing have the potential to cause disease. Many real-time, molecular-based assays have been developed to detect Enterohemorrhagic E. coli (EHEC) including E. coli O157:H7. However, no assay currently exists to detect the eae gene present in E. coli O157:H7 and other EHEC strains with a confirmed positive or negative result in less than 12 h. Raw beef food products (raw ground beef and raw boneless beef) at 25 and 375 g samples and bovine fecal samples at 2 g were inoculated with 10(1), 10(3), 10(4), and 10(5) organisms of E. coli O157:H7 to test the sensitivity of this assay. Fourteen different foodborne bacteria (including E. coli O157:H7) and 19 various E. coli strains, obtained from the United States Department of Agriculture-Agricultural Research Service (USDA-ARS) were tested for specificity. E. coli O157:H7 was detected at the level of 10(1) organisms in both 25 and 375 g samples of raw ground and raw boneless beef products as well as 2 g samples of bovine feces after pre enrichment and concentration. None of the 14 foodborne bacteria screened for cross-reactivity was detected. All USDA E. coli strains confirmed to contain the eae gene were detected. PMID- 15797824 TI - Detection of Mycobacterium avium subspecies paratuberculosis in free-ranging bison (Bison bison) by PCR. AB - Bacterial culture is the 'gold standard' for detecting Mycobacterium avium subspecies paratuberculosis (MAP) infection, but is time consuming, laborious, and recovery of organism varies with species of animal tested. PCR has been used for detection of MAP DNA in feces and tissues. We used PCR to detect MAP DNA isolated from tissues from 25 free-ranging North American bison (Bison bison), each with clinical signs compatible with Johne's disease. We report the performance of PCR to detect MAP DNA in both frozen and paraffin-embedded ileum, jejunum, and ileocecal lymph node samples collected at the time of slaughter. Specific oligonucleotide primers for PCR amplification were derived from 16S rRNA sequence M. avium subspecies (MAs) and insertion elements IS1245 (MAs avium), IS901 (MAs avium), IS900 (MAP), and hspX (MAP). Genomic DNA samples were prepared three different ways; crude DNA from frozen tissues, crude DNA from paraffin embedded tissues, and purified DNA from paraffin-embedded tissues. An animal was considered infected if MAP DNA was detected in at least two separate tissues using the IS900 primer set. Using these criteria, 25 of 25 bison tested were positive for MAP. The data indicate that these free-ranging bison have been infected by MAP. PMID- 15797825 TI - Introduction: new aspects of cellular sterol metabolism and transport. PMID- 15797826 TI - Sterol homeostasis in the budding yeast, Saccharomyces cerevisiae. AB - Lipid related diseases, such as obesity, type 2 diabetes, and atherosclerosis are epidemics in developed civilizations. A common underlying factor among these syndromes is excessive subcellular accumulation of lipids such as cholesterol and triglyceride. The homeostatic events that govern these metabolites are understood to varying degrees of sophistication. We describe here the utilization of a genetically powerful model organism, budding yeast, to identify and characterize novel aspects of sterol and lipid homeostasis. PMID- 15797827 TI - Caveolin, cholesterol, and lipid bodies. AB - In mammalian cells a complex interplay regulates the distribution of cholesterol between intracellular membrane compartments. One important aspect of cholesterol regulation is intracellular cholesterol storage in neutral lipid storage organelles called lipid droplets or lipid bodies (LBs). Recent work has thrust the LB into the limelight as a complex and dynamic cellular organelle. LBs play a crucial role in maintaining the cellular levels of cholesterol by regulating the interplay between lipid storage, hydrolysis and trafficking. Studies of caveolins, caveolar membrane proteins linked to lipid regulation, are providing new insights into the role of LBs in regulating cholesterol balance. PMID- 15797828 TI - Requirement of sterols in the life cycle of the nematode Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans represents an excellent model for studying many aspects of sterol function on the level of a whole organism. Recent studies show that especially two processes in the life cycle of the worm, dauer larva formation and molting, depend on sterols. In both cases, cholesterol or its derivatives seem to act as hormones rather than being structural components of the membrane. Investigations on C. elegans could provide information on the etiology of human diseases that display defects in the transport or metabolism of sterols. PMID- 15797829 TI - Molecular mechanisms of cholesterol absorption and transport in the intestine. AB - Many enzymes and transport proteins participate in cholesterol absorption. This review summarizes recent results on several proteins that are important for each step of the cholesterol absorption pathway, including the important roles of: (i) pancreatic triglyceride lipase (PTL), carboxyl ester lipase (CEL), and ileal bile acid transporter in determining the rate of cholesterol absorption; (ii) ATP binding cassette (ABC) transporters and the Niemann-Pick C-1 like-1 (NPC1L1) protein as intestinal membrane gatekeepers for cholesterol efflux and influx; and (iii) intracellular membrane vesicles and transport proteins in lipid trafficking through intracellular compartments prior to lipoprotein assembly and secretion to plasma circulation. PMID- 15797831 TI - Programmed cell death in development. PMID- 15797830 TI - Cholesterol homeostasis in neurons and glial cells. AB - Cholesterol is highly enriched in the brain compared to other tissues. Essentially all cholesterol in the brain is synthesized endogenously since plasma lipoproteins are unable to cross the blood-brain barrier. Cholesterol is transported within the central nervous system in the form of apolipoprotein E containing lipoprotein particles that are secreted mainly by glial cells. Cholesterol is excreted from the brain in the form of 24-hydroxycholesterol. Apolipoprotein E and cholesterol have been implicated in the formation of amyloid plaques in Alzheimer's disease. In addition, the progressive neurodegenerative disorder Niemann-Pick C disease is characterized by defects in intracellular trafficking of cholesterol. PMID- 15797832 TI - From oogenesis through gastrulation: developmental regulation of apoptosis. AB - Apoptosis is a mechanism employed by multicellular organisms throughout development as a means of eliminating damaged or otherwise unwanted cells. From oogenesis through fertilization and gastrulation, organisms use an array of cell- and tissue-specific mechanisms to regulate the apoptotic program in response to stress or developmental cues. Since cell death regulation is tightly interwoven with cell cycle and checkpoint controls, and embryos of the fly, fish and frog exhibit unique embryonic cell cycle regulation, it is of great interest to understand how early embryos coordinate these cellular functions. PMID- 15797833 TI - Genetic control of programmed cell death in Drosophila melanogaster. AB - Apoptosis is a genetically controlled form of cell death that is an important feature of animal development and homeostasis. The genes involved in the control and execution of apoptosis are conserved throughout evolution. However, the actual molecular mechanisms used by these genes vary from species to species. In this review, we focus on the genetic components of apoptosis in the fruit fly Drosophila melanogaster, and compare their mode of action to the one employed by the homologous genes in mammals. We also cover recent advances that show that apoptotic genes have a requirement in processes other than apoptosis. PMID- 15797834 TI - Mechanisms of steroid-triggered programmed cell death in Drosophila. AB - Studies in Drosophila have provided a detailed understanding of how programmed cell death is regulated by steroid hormones during development. This work has defined a two-step hormone-triggered regulatory cascade that results in the coordinate induction of central players in the death pathway, including the reaper and hid death activators, the Apaf-1 ortholog dark, and the dronc apical caspase gene. Recent transcriptional profiling studies have identified many new players in this pathway. In addition, genetic studies are providing new insights into the control of autophagic cell death and revealing how this response is related to, but distinct from, apoptosis. PMID- 15797835 TI - Programmed cell death in the germline. AB - In many organisms, programmed cell death of germ cells is required for normal development. This often occurs through highly conserved events including the transfer of vital cellular material to the growing gametes following death of neighboring cells. Germline cell death also plays a role in such diverse processes as removal of abnormal or superfluous cells at certain checkpoints, establishment of caste differentiation, and individualization of gametes. This review focuses on the cell death events that occur during gametogenesis in both vertebrates and invertebrates. It also examines the signals and machinery that initiate and carry out these germ cell deaths. PMID- 15797836 TI - Programmed cell death in the embryonic vertebrate limb. AB - The developing limb bud provides one of the best examples in which programmed cell death exerts major morphogenetic functions. In this work, we revise the distribution and the developmental significance of cell death in the embryonic vertebrate limb and its control by the BMP signalling pathway. In addition, paying special attention to the interdigital apoptotic zones, we review current data concerning the intracellular death machinery implicated in mesodermal limb apoptosis. PMID- 15797837 TI - Programmed cell death during amphibian metamorphosis. AB - The death of different types of cells occurs in regressing or remodeling organs to transform from a tadpole to a frog in both temporally and spatially regulated manners during amphibian metamorphosis. This morphological change is drastic and visible with the naked eye. This review summarizes our current understanding of the basic mechanism of the cell death during the metamorphosis. It focuses in particular on the tail resorption and the remodeling of intestine and skin where programmed cell death is executed by thyroid hormone-signaling through the cell autonomous response (suicide) and the degradation of the extracellular matrix (murder). PMID- 15797838 TI - Expanding roles of programmed cell death in mammalian neurodevelopment. AB - Programmed cell death is an orchestrated form of cell death in which cells are actively involved in their own demise. During neural development in mammals, many progenitor cells, immature cells or differentiated cells undergo the most clearly characterized type of cell death, apoptosis. Several pathways of apoptosis have been linked to neural development, but according to the numerous and striking phenotypes observed when apoptotic genes are inactivated, the mitochondrial death route is the most important pathway in this context. Here, we discuss the relative importance of pro-growth/pro-death factors in the control of neural tissue development. We also discuss the impact of studying programmed cell death in development in order to better understand the basis of several human diseases and embryonic defects of the nervous system. PMID- 15797839 TI - Clearance of apoptotic cells in Caenorhabditis elegans. AB - Programmed cell death, or apoptosis, is a genetically controlled process of cell suicide that is a common fate during an animal's life. In metazoans, apoptotic cells are rapidly removed from the body through the process of phagocytosis. Genetic analyses probing the mechanisms controlling the engulfment of apoptotic cells were pioneered in the nematode Caenorhabditis elegans. So far, at least seven genes have been identified that are required for the recognition and engulfment of apoptotic cells and have been shown to function in two partially redundant signaling pathways. Molecular characterization of their gene products has lead to the finding that similar genes act to control the same processes in other organisms, including mammals. In this paper, we review these exciting findings in C. elegans and discuss their implications in understanding the clearance of apoptotic cells in mammals. PMID- 15797840 TI - Determination of alveolar-capillary O2 partial pressure gradient by using 15NO. AB - We propose an approach for determining the alveolar-mean capillary oxygen (O(2)) partial pressure gradient to evaluate the efficiency of O(2) equilibration between alveolar space and pulmonary capillary blood. For this purpose, measurements of the pulmonary [(15)N]nitric oxide diffusing capacity are to be interpolated into the recording of O(2) consumption. We expect the O(2) partial pressure gradient amounting to 3.3 mmHg for breathing room air at rest, a third of that commonly given. The simplicity of our method allows its application to children or even artificially ventilated patients. Therefore, it will enable a new insight into pulmonary O(2) equilibration. PMID- 15797841 TI - A direct nitric oxide gas delivery system for bacterial and mammalian cell cultures. AB - Nitric oxide (NO) is the smallest known gaseous signaling molecule released by mammalian and plant cells. To investigate the pathophysiologic role of exogenous NO gas (gNO) in bacterial and mammalian cell cultures, a validated in vitro delivery method is required. The system should be able to deliver gNO directly to bacterial and/or cell cultures in a continuous, predictable, and reproducible manner over a long period of time (days). To accomplish this, a gas delivery system was designed to provide optimal growth conditions for bacteria and/or mammalian cells. Parameters for cell exposure, such as concentration of gNO, nitrogen dioxide (NO(2)), oxygen (O(2)), temperature, and relative humidity (RH) were continuously monitored and evaluated. Uptake of gNO into various media was monitored by measuring the nitrite concentration using the Griess reagent technique. A selection of standard growth media [saline, tryptic soy broth (TSB), Middlebrook 7H9 (MB 7H9), and Dulbecco's modified Eagle's medium (DMEM)] exposed to various concentrations of gNO revealed a steady and consistent transfer of gNO into the aqueous phase over a 48-h period. Validation of optimal growth conditions within the device, as compared to a conventional incubator, were accomplished by growing and observing viability of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and human fibroblast cultures in the absence of gNO. These results indicate that an optimal growth environment for the above tested cells was accomplished inside the proposed delivery system. Dose-dependent toxicological data revealed a significant bacteriostatic effect on P. aeruginosa and S. aureus with continuous exposure to 80 ppm gNO. No toxic effects were observed on dermal fibroblast proliferation at concentrations up to 400 ppm gNO for 48 h. In conclusion, the designed gNO exposure system is capable of supporting cellular viability for a representative range of prokaryote and eukaryotic cells. The exposure system is also capable of obtaining toxicological data. Therefore, the proposed device can be utilized to continuously expose cells to various levels of gNO for up to 72 h to study the in vitro effects of gNO therapy. PMID- 15797842 TI - A systematic review of nitric oxide donors and L-arginine in experimental stroke; effects on infarct size and cerebral blood flow. AB - BACKGROUND: Nitric oxide (NO) is a candidate treatment for acute ischaemic stroke, however published studies in experimental stroke have given conflicting results. METHODS: We performed a systematic review of published controlled studies of L-arginine (the precursor for NO) and NO donors in experimental stroke. Data were analysed using the Cochrane Collaboration Review Manager software. Standardised mean difference (SMD) and 95% confidence intervals (95% CI) were calculated. RESULTS: Altogether, 25 studies(s) were identified. L Arginine and NO donors reduced total cerebral infarct volume in permanent (SMD 1.21, 95% CI -1.69 to -0.73, p < 0.01, s = 10) and transient models of ischaemia (SMD -0.78, 95% CI -1.21 to -0.35, p < 0.01, s = 7). Drug administration increased cortical CBF in permanent (SMD +0.86, 95% CI 0.52-1.21, p < 0.01, s = 8) but not transient models (SMD +0.34, 95% CI -0.02 to 0.70, p = 0.07, s = 4). CONCLUSIONS: Administration of NO in experimental stroke reduces stroke lesion volume in permanent and transient models. This may be mediated, in part, by increased cerebral perfusion in permanent models. These data support clinical trials in stroke patients, although the presence of a narrow therapeutic time window may be a limiting factor. PMID- 15797843 TI - PGE2 enhances cytokine-elicited nitric oxide production in mouse cortical collecting duct cells. AB - It has been documented that arginine vasopressin (AVP) and prostaglandin E(2) (PGE(2)) regulate water reabsorption in renal tubular cells. The present study was attempted to delineate the downstream signaling of AVP and PGE(2) in a cortical collecting duct cell line (M-1 cell). Using RT-PCR, we detected mRNA for V2 and VACM-1 but not for V1a and AII/AVP receptors of AVP. Furthermore, neither AVP nor V2 receptor agonist and antagonist alter cellular cAMP. These together with unchanged cellular Ca(2+) by AVP suggested that AVP pathway was not operating in M-1 cells. All four classical PGE(2) receptors with EP3 and EP4 as the most prominent were detected in M-1 cells. PGE(2), 11-deoxy-PGE(1) (EP2 and EP4 agonist), and 17-phenyl-trinor-PGE(2) (EP1 agonist) increased cellular concentration of cAMP. There was no effect of PGE(2) or EP1 agonist on cellular Ca(2+). These findings provide evidence of the involvement of PGE(2) cascade in M 1 cells. M-1 cells were capable of synthesizing nitric oxide (NO). Although individual cytokines did not affect NO production, a mixture of tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma elevated NO concentration to 4.5-fold of the control. Addition of PGE(2) and db-cAMP to the cytokine mixture further increased NO production to 7.0- and 9.8-fold, respectively, of that seen in non-treated cells. PGE(2) or db-cAMP alone, however, had no effect on NO production. The results of the study led us to speculate that enhanced production of cAMP via PGE(2) signaling pathway in M-1 cells could either stimulate or attenuate water reabsorption in renal tubule. While an increase in cAMP alone may enhance water reabsorption, a concomitant increase in cAMP and cytokines may inhibit water reabsorption in renal tubule. PMID- 15797845 TI - Consistent interethnic differences in the distribution of clinically relevant endothelial nitric oxide synthase genetic polymorphisms. AB - A maldistribution of endothelial nitric oxide synthase (eNOS) genetic variants may explain differences in NO-mediated effects and response to drugs among black and white subjects. While interethnic differences in the distribution of eNOS genetic variants exist in the American population, it is not known whether such interethnic differences exist in other populations. To test this possibility, we examined the distribution of genetic variants of three clinically relevant eNOS polymorphisms (T(-786)C in the promoter, the VNTR in intron 4, and the Glu298Asp variant in exon 7) in 136 black and 154 white subjects from a Brazilian population, which is very heterogeneous. We also estimated the haplotype frequency and evaluated associations between these variants. The Asp298 variant was more common in whites (32.8%) than in blacks (15.1%) (P < 0.004). Similarly, the C(-786) variant was more common in whites (41.9%) than in blacks (19.5%) (P < 0.0004). However, the 4a variant was more common in blacks (32.0%) than in whites (17.9%) (P < 0.003). The most common predicted haplotype in both ethnic groups combined only wild-type variants. While the second most common haplotype in blacks includes the variant 4a and the wild-type variants for the remaining polymorphisms, the second most common haplotype in whites includes the variants Asp298 and C(-786) and the wild-type variant for polymorphism in intron 4. The marked interethnic differences that we found in Brazilians are very similar to those previously reported in Americans. These findings strongly suggest a consistent difference in the distribution of eNOS genetic variants in blacks compared with whites and indicate that the interethnic differences do not vary with geographic origin. PMID- 15797844 TI - Korea red ginseng water extract increases nitric oxide concentrations in exhaled breath. AB - Panax ginseng is well known to enhance the release of nitric oxide (NO) from endothelial cells of the rat aorta and to reduce blood pressure in animals. In this study, we investigated the effects of water extract of Korea red ginseng (KRG) on NO concentration levels in the exhaled breath, blood pressure, and heart rate of human volunteers. We also are interested in whether NO levels in exhaled breath are increased by KRG extract, and correlated with blood pressure and heart rate. Twelve healthy, non-smoking male volunteers were recruited for this study. A single administration of KRG water extract (500 mg/50 kg) increased NO levels in exhaled breath, and concomitantly decreased mean blood pressure and heart rate. The correlation value between NO levels and heart rate (r = 0.94), and the correlation value between NO levels and heart rate (r = 0.84) are significant (P < 0.01). Linear regression analysis shows the clear conversed correlation between NO levels and blood pressure as well as heart rate. Therefore, present data suggest that KRG may be useful for the treatment of hypertension and pulmonary vascular obstruction. PMID- 15797846 TI - Is there a difference between evaluation and research? PMID- 15797848 TI - Diabetes and osteoporotic fractures: the role of the diabetes educator. AB - The purpose of this article is to present information that will assist the diabetes educator in screening patients with diabetes for risk of osteoporosis and fracture, to offer appropriate treatment options for patients, and to identify potential referrals to other providers for patients with diabetes and increased risk of fracture. PMID- 15797849 TI - IT'S MINE: Initiating Treatment Success--My Individualized Nutrition and Exercise Plan. AB - The authors' class structure incorporates individual meal and exercise planning in the group setting. This study was conducted to (1) determine if class participants can create their own meal and exercise plans, (2) determine if class participants feel they will be able to adhere to their meal and exercise plans, and (3) assess adherence to the plans over time. Subjects were recruited from the classes offered at the Joslin Diabetes Center. Following completion of class, patients completed the evaluation questionnaire. Two and 6 months later, study participants received follow-up questionnaires by phone or mail. Among respondents, 63% were able to determine their own carbohydrate goals, with 95.9% indicating they could adhere to the plan, and 82.8% felt the plan would be easier than previous ones. At 2 and 6 months, respectively, 89% and 92% of the participants felt they were following the meal plan either some or most of the time. One hundred percent of the respondents were able to determine their own exercise plan, with 98% indicating they could adhere to the plan, and 85.7% felt the new plan would be easier than previous ones. At 2 and 6 months, respectively, 70% and 73% felt they were following their exercise plan either some or most of the time. Individualized meal and exercise plans can be successfully created in a group setting. PMID- 15797850 TI - The Spoken Knowledge in Low Literacy in Diabetes scale: a diabetes knowledge scale for vulnerable patients. AB - PURPOSE: The purpose of this study was to develop and validate a new knowledge scale for patients with type 2 diabetes and poor literacy: the Spoken Knowledge in Low Literacy patients with Diabetes (SKILLD). METHODS: The authors evaluated the 10-item SKILLD among 217 patients with type 2 diabetes and poor glycemic control at an academic general medicine clinic. Internal reliability was measured using the Kuder-Richardson coefficient. Performance on the SKILLD was compared to patient socioeconomic status, literacy level, duration of diabetes, and glycated hemoglobin (A1C). RESULTS: Respondents' mean age was 55 years, and they had diabetes for an average of 8.4 years; 38% had less than a sixth-grade literacy level. The average score on the SKILLD was 49%. Less than one third of patients knew the signs of hypoglycemia or the normal fasting blood glucose range. The internal reliability of the SKILLD was good (0.72). Higher performance on the SKILLD was significantly correlated with higher income (r = 0.22), education level (r = 0.36), literacy status (r = 0.33), duration of diabetes (r = 0.30), and lower A1C (r = -0.16). When dichotomized, patients with low SKILLD scores (< or = 50%) had significantly higher A1C (11.2% vs 10.3%, P < .01). This difference remained significant when adjusted for covariates. CONCLUSION: The SKILLD demonstrated good internal consistency and validity. It revealed significant knowledge deficits and was associated with glycemic control. The SKILLD represents a practical scale for patients with diabetes and low literacy. PMID- 15797851 TI - Implementing the chronic care model for improvements in diabetes care and education in a rural primary care practice. AB - PURPOSE: The purpose of this pilot study was to determine the impact of implementing elements of the chronic care model (CCM; decision support, self management, and delivery system redesign) on providers' diabetes care practices and patient outcomes in a rural practice setting. METHODS: In this pilot study, 104 patients with type 2 diabetes and 6 providers in a rural primary care practice were involved in an intervention that included a certified diabetes educator (CDE) who educated and supported providers on diabetes management and adherence to the American Diabetes Association (ADA) Standards of Care over the year of the project. The CDE also provided diabetes self-management education (DSME) at the office site for 29 of the 104 patients who received their care in the practice. The following variables were evaluated: provider perceived barriers to care and adherence to ADA standards of care and patient A1C, blood pressure, cholesterol, knowledge, and empowerment levels. RESULTS: Provider adherence to ADA Standards of Care increased significantly across all process measures. Patients who received DSME at point of service in the primary care practice setting gained improvements in knowledge, empowerment, A1C, and high-density lipoprotein cholesterol levels. CONCLUSIONS: Implementing systems to support decision support, self-management education, and delivery system redesign has a positive influence on practices and patient outcomes in outlying rural communities. PMID- 15797852 TI - Barriers to A1C testing among a managed care population. AB - PURPOSE: The purpose of this study is to explore reasons adults with diabetes do not receive at least 2 A1C tests per year as recommended by the American Diabetes Association (ADA). METHODS: ConnectiCare, a regional managed care company based in Farmington, Connecticut, identified adult members with diabetes who did not have a medical claim for an A1C laboratory test from their physician. A questionnaire was sent to 740 randomly selected members asking them to report the number of A1C tests they received in the past 12 months and reasons for not receiving the number of tests recommended by the ADA. After sending an automated telephone reminder to nonrespondents, a 26% (n = 192) response rate was achieved. RESULTS: Thirty-three percent of respondents (n = 63) reported having diabetes and receiving fewer than 2 A1C tests in the past year. Respondents were equally divided between men and women, with a mean age of 58 years. The primary reasons given for not obtaining at least 2 A1C tests as recommended by the ADA were that respondents were unaware that the test is recommended (49%), not informed of the need for the test by their physician (38%), never heard of the A1C test (33%), and not seen regularly by their physician (19%). CONCLUSIONS: Diabetes self management education remains an important means of encouraging adherence to important ADA recommendations such as regular A1C testing. Barriers to A1C testing can be addressed in multiple settings, including individual and group education, disease management programs, and physician education. PMID- 15797853 TI - Patient adherence improves glycemic control. AB - PURPOSE: The purpose of this study was to assess the influence of appointment keeping and medication adherence on HbA1c. METHODS: A retrospective evaluation was performed in 1560 patients with type 2 diabetes who presented for a new visit to the Grady Diabetes Clinic between 1991 and 2001 and returned for a follow-up visit and HbA1c after 1 year of care. Appointment keeping was assessed by the number of scheduled intervening visits that were kept, and medication adherence was assessed by the percentage of visits in which self-reported diabetes medication use was as recommended at the preceding visit. RESULTS: The patients had an average age of 55 years, body mass index (BMI) of 32 kg/m2, diabetes duration of 4.6 years, and baseline HbA1c of 9.1%. Ninety percent were African American, and 63% were female. Those who kept more intervening appointments had lower HbA1c levels after 12 months of care (7.6% with 6-7 intervening visits vs 9.7% with 0 intervening visits). Better medication adherence was also associated with lower HbA1c levels after 12 months of care (7.8% with 76%-100% adherence). After adjusting for age, gender, race, BMI, diabetes duration, and diabetes therapy in multivariate linear regression analysis, the benefits of appointment keeping and medication adherence remained significant and contributed independently; the HbA1c was 0.12% lower for every additional intervening appointment that was kept (P = .0001) and 0.34% lower for each quartile of better medication adherence (P = .0009). CONCLUSION: Keeping more appointments and taking diabetes medications as directed were associated with substantial improvements in HbA1c. Efforts to enhance glycemic outcomes should include emphasis on these simple but critically important aspects of patient adherence. PMID- 15797854 TI - Recruitment and retention of rural African Americans in diabetes research: lessons learned. AB - PURPOSE: The purpose of this article was to describe lessons learned about recruitment and retention of rural African Americans from published literature, the authors' research, and research experience. Two rural, community-based research studies with African Americans with diabetes are used to illustrate different issues and strategies in recruitment and retention. METHODS: Relevant MEDLINE articles and clinical studies were reviewed, and the design, implementation, and results of the 2 community-based studies were evaluated. Information from the literature, research results, and sample selection, participation, and attrition experiences were synthesized to determine effective approaches for recruitment and retention. RESULTS: Research funding, design, and implementation; recruitment methods; culturally competent approaches; caring, trusting provider-patient relationships; incentives; follow-up; and factors in the rural environment emerged as important issues influencing recruitment and retention. CONCLUSION: Recruitment and retention of African Americans in rural diabetes research is a significant challenge, and adequate funding should be sought early in the research process. Culturally competent approaches; caring, trusting relationships; incentives; and follow-up are important concepts in successful recruitment, participation, and retention of African Americans. The lessons learned may be applicable to the more widespread issue of recruitment and retention of rural African Americans in diabetes education programs. PMID- 15797855 TI - Heparan sulfate targets the HIV-1 envelope glycoprotein gp120 coreceptor binding site. AB - Human immunodeficiency virus (HIV) attachment to host cells is a multi-step process that involves interaction of the viral envelope gp120 with the primary receptor CD4 and coreceptors. HIV gp120 also binds to other cell surface components, including heparan sulfate (HS), a sulfated polysaccharide whose wide interactive properties are exploited by many pathogens for attachment and concentration at the cell surface. To analyze the structural features of gp120 binding to HS, we used soluble CD4 to constrain gp120 in a specific conformation. We first found that CD4 induced conformational change of gp120, dramatically increasing binding to HS. We then showed that HS binding interface on gp120 comprised, in addition to the well characterized V3 loop, a CD4-induced epitope. This epitope is efficiently targeted by nanomolar concentrations of size-defined heparin/HS-derived oligosaccharides. Because this domain of the protein also constitutes the binding site for the viral coreceptors, these results support an implication of HS at late stages of the virus-cell attachment process and suggest potential therapeutic applications. PMID- 15797856 TI - Deficiency of the G-protein alpha-subunit G(s)alpha in osteoblasts leads to differential effects on trabecular and cortical bone. AB - The G-protein alpha-subunit G(s)alpha is required for the intracellular cAMP responses to hormones and other agonists. G(s)alpha is known to mediate the cAMP response to parathyroid hormone and other hormones and cytokines in bone and cartilage. To analyze the in vivo role of G(s)alpha signaling in osteoblasts, we developed mice with osteoblast/osteocyte-specific G(s)alpha deficiency (BGsKO) by mating G(s)alpha-floxed mice with collagen Ialpha1 promoter-Cre recombinase transgenic mice. Early skeletal development was normal in BGsKO mice, because formation of the initial cartilage template and bone collar was unaffected. The chondrocytic zones of the growth plates also appeared normal in BGsKO mice. BGsKO mice had a defect in the formation of the primary spongiosa with reduced immature osteoid (new bone formation) and overall length, which led to reduced trabecular bone volume. In contrast, cortical bone was thickened with narrowing of the bone marrow cavity. This was probably due to decreased cortical bone resorption, because osteoclasts were markedly reduced on the endosteal surface of cortical bone. In addition, the expression of alkaline phosphatase, an early osteoblastic differentiation marker, was normal, whereas the expression of the late osteoblast differentiation markers osteopontin and osteocalcin was reduced, suggesting that the number of mature osteoblasts in bone is reduced. Expression of the osteoclast stimulating factor receptor activator of NF-kappaB ligand was also reduced. Overall, our findings have similarities to parathyroid hormone null mice and confirm that the differential effects of parathyroid hormone on trabecular and cortical bone are primarily mediated via G(s)alpha in osteoblasts. Osteoblast specific G(s)alpha deficiency leads to reduced bone turnover. PMID- 15797857 TI - Characterization of heparin affin regulatory peptide signaling in human endothelial cells. AB - Heparin affin regulatory peptide (HARP) is an 18-kDa secreted growth factor that has a high affinity for heparin and a potent role on tumor growth and angiogenesis. We have previously reported that HARP is mitogenic for different types of endothelial cells and also affects cell migration and differentiation (12). In this study we examined the signaling pathways involved in the migration and tube formation on matrigel of human umbilical vein endothelial cells (HUVEC) induced by HARP. We report for the first time that receptor-type protein-tyrosine phosphatase beta/zeta (RPTPbeta/zeta), which is a receptor for HARP in neuronal cell types, is also expressed in HUVEC. We also document that HARP signaling through RPTPbeta/zeta leads to activation of Src kinase, focal adhesion kinase, phosphatidylinositol 3-kinase, and Erk1/2. Sodium orthovanadate, chondroitin sulfate-C, PP1, wortmannin, LY294002, and U0126 inhibit HARP-mediated signaling and HUVEC migration and tube formation. In addition, RPTPbeta/zeta suppression using small interfering RNA technology interrupts intracellular signals and HUVEC migration and tube formation induced by HARP. These results establish the role of RPTPbeta/zeta as a receptor of HARP in HUVEC and elucidate the HARP signaling pathway in endothelial cells. PMID- 15797858 TI - Mutation in apolipoprotein B associated with hypobetalipoproteinemia despite decreased binding to the low density lipoprotein receptor. AB - Mutations in apolipoprotein B (APOB) may reduce binding of low density lipoprotein (LDL) to the LDL receptor and cause hypercholesterolemia. We showed that heterozygotes for a new mutation in APOB have hypobetalipoproteinemia, despite a reduced binding of LDL to the LDL receptor. APOB R3480P heterozygotes were identified among 9,255 individuals from the general population and had reduced levels of apoB-containing lipoproteins. Most surprisingly, R3480P LDL bound with lower affinity to the LDL receptor than non-carrier LDL in vitro, and these results were confirmed by turnover studies of LDL in vivo. In very low density lipoprotein (VLDL) turnover studies, the amount of VLDL converted to LDL in R3480P heterozygotes was substantially reduced, suggesting that this was the explanation for the hypobetalipoproteinemia observed in these individuals. Our findings emphasized the importance of combining in vitro studies with both human in vivo and population-based studies, as in vitro studies often have focused on very limited aspects of complex mechanisms taken out of their natural context. PMID- 15797860 TI - An NIH career: from bedside to basic research and back. PMID- 15797859 TI - p38 kinase-mediated transactivation of the epidermal growth factor receptor is required for dedifferentiation of renal epithelial cells after oxidant injury. AB - Renal proximal tubular cell (RPTC) dedifferentiation is thought to be a prerequisite for regenerative proliferation and migration after renal injury. However, the specific mediators and the mechanisms that regulate RPTC dedifferentiation have not been elucidated. Because epidermal growth factor (EGF) receptor activity is required for recovery from acute renal failure, we examined the role of the EGF receptor in dedifferentiation and the mechanisms of EGF receptor transactivation in primary cultures of RPTCs after oxidant injury. Exposure of confluent RPTCs to H2O2 resulted in 40% cell death, and surviving RPTCs acquired a dedifferentiated phenotype (e.g. elongated morphology and vimetin expression). The EGF receptor, p38, Src, and MKK3 were activated after oxidant injury and inhibition of the EGF receptor or p38 with specific inhibitors (AG1478 and SB203580, respectively) blocked RPTC dedifferentiation. Treatment with SB203580 or adenoviral overexpression of dominant negative p38alpha or its upstream activator, MKK3, inhibited EGF receptor phosphorylation induced by oxidant injury, whereas AG1478 had no effect on p38 phosphorylation. Inhibition of Src with 4-amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]-pyrimidine (PP1) blocked MKK3 and p38 activation, and inhibition of MKK3 blocked p38 activation. In addition, inactivation of Src, MKK3, p38, or the EGF receptor blocked tyrosine phosphorylation of beta-catenin, a key signaling intermediate that is involved in the epithelial-mesenchymal transition and vimentin expression. These results reveal that p38 mediates EGF receptor activation after oxidant injury; that Src activates MMK3, which, in turn, activates p38; and that the EGF receptor signaling pathway plays a critical role in RPTC dedifferentiation. PMID- 15797861 TI - Identification of key amino acids responsible for the substantially higher affinities of human type 1 3beta-hydroxysteroid dehydrogenase/isomerase (3beta HSD1) for substrates, coenzymes, and inhibitors relative to human 3beta-HSD2. AB - The human type 1 (placenta, breast tumors, and prostate tumors) and type 2 (adrenals and gonads) isoforms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD1 and 3beta-HSD2) are encoded by two distinct genes that are expressed in a tissue-specific pattern. Our recent studies have shown that His156 contributes to the 14-fold higher affinity that 3beta-HSD1 exhibits for substrate and inhibitor steroids compared with human 3beta-HSD2 containing Tyr156 in the otherwise identical catalytic domain. Our structural model of human 3beta-HSD localizes His156 or Tyr156 in the subunit interface of the enzyme homodimer. The model predicts that Gln105 on one enzyme subunit has a higher probability of interacting with His156 on the other subunit in 3beta-HSD1 than with Tyr156 in 3beta-HSD2. The Q105M mutant of 3beta-HSD1 (Q105M1) shifts the Michaelis-Menten constant (Km) for 3beta-HSD substrate and inhibition constants (Ki) for epostane and trilostane to the much lower affinity profiles measured for wild-type 3beta HSD2 and H156Y1. However, the Q105M2 mutant retains substrate and inhibitor kinetic profiles similar to those of 3beta-HSD2. Our model also predicts that Gln240 in 3beta-HSD1 and Arg240 in 3beta-HSD2 may be responsible for the 3-fold higher affinity of the type 1 isomerase activity for substrate steroid and cofactors. The Q240R1 mutation increases the isomerase substrate Km by 2.2-fold to a value similar to that of 3beta-HSD2 isomerase and abolishes the allosteric activation of isomerase by NADH. The R240Q2 mutation converts the isomerase substrate, cofactor, and inhibitor kinetic profiles to the 4-14-fold higher affinity profiles of 3beta-HSD1. Thus, key structural reasons for the substantially higher affinities of 3beta-HSD1 for substrates, coenzymes, and inhibitors have been identified. These structure and function relationships can be used in future docking studies to design better inhibitors of the 3beta-HSD1 that may be useful in the treatment of hormone-sensitive cancers and preterm labor. PMID- 15797862 TI - A specific interaction between muskelin and the cyclin-dependent kinase 5 activator p39 promotes peripheral localization of muskelin. AB - Previous studies implicate cyclin-dependent kinase 5 in cell adhesion and migration of epithelial cells of the cornea and lens. To explore molecular interactions underlying these functions, we performed yeast two-hybrid screening of an embryonic rat lens library for proteins that interact with cyclin-dependent kinase 5 and its regulators, p35 and p39. This screen identified a specific interaction between p39 and muskelin, an intracellular protein known to affect cytoskeletal organization in adherent cells. Immunohistochemistry detected muskelin in the developing lens and in other tissues, including brain and muscle. Glutathione S-transferase pull-down experiments and co-immunoprecipitations confirmed the specificity of the p39-muskelin interaction. Deletion analysis of p39 showed that muskelin binds to the p39 C terminus, which contains a short insertion (amino acids 329-366) absent from p35. Similar analysis of muskelin mapped the interaction with p39 to the fifth kelch repeat. Co-expression of p39 and muskelin in COS1 cells or lens epithelial cells altered the intracellular localization of muskelin, recruiting it to the cell periphery. These findings demonstrate a novel interaction between muskelin and the cyclin-dependent kinase 5 activator p39 and suggest that p39 may regulate the subcellular localization of muskelin. PMID- 15797863 TI - Presenilin-1 D257A and D385A mutants fail to cleave Notch in their endoproteolyzed forms, but only presenilin-1 D385A mutant can restore its gamma secretase activity with the compensatory overexpression of normal C-terminal fragment. AB - The enzyme gamma-secretase is involved in the cleavage of several type I membrane proteins, such as Notch 1 and amyloid precursor protein. Presenilin-1 (PS-1) is one of the critical integral membrane protein components of the gamma-secretase complex and is processed endoproteolytically, generating N- and C-terminal fragments (NTF and CTF, respectively). PS-1 is also known to incorporate into a high molecular weight complex by binding to other gamma-secretase components such as Nicastrin, Aph-1, and Pen-2. Mutations on PS-1 can alter the effects of gamma secretase on its many substrates to different extents. Here, we showed that PS-1 mutants have a different activity for Notch cleavage, which depended on the PS-1 mutation site. We demonstrated that defective PS-1 mutants located in CTF, i.e. D385A and C410Y, could restore their gamma-secretase activities with the compensatory overexpression of wild type CTF or of minimal deleted CTF (amino acids 349-467). However, the defective PS-1 D257A mutant could not restore their gamma-secretase activities with the compensatory overexpression of wild type NTF. In comparison, both D257A NTF and D385A CTF could abolish the gamma-secretase activity of wild type and pathogenic PS-1 mutants. We also showed that PS-1 NTF but not CTF forms strong high molecular weight aggregates in SDS-PAGE. Taken together, results have shown that NTF and CTF integrate differently into high molecular weight aggregates and that PS-1 Asp-257 and Asp-385 have different accessibilities in their unendoproteolyzed conformation. PMID- 15797864 TI - KCNQ-like potassium channels in Caenorhabditis elegans. Conserved properties and modulation. AB - The human KCNQ gene family encodes potassium channels linked to several genetic syndromes including neonatal epilepsy, cardiac arrhythmia, and progressive deafness. KCNQ channels form M-type potassium channels, which are critical regulators of neuronal excitability that mediate autonomic responses, pain, and higher brain function. Fundamental mechanisms of the normal and abnormal cellular roles for these channels may be gained from their study in simple model organisms. Here we report that a multigene family of KCNQ-like channels is present in the nematode, Caenorhabditis elegans. We show that many aspects of the functional properties, tissue expression pattern, and modulation of these C. elegans channels are conserved, including suppression by the M1 muscarinic receptor. We also describe a conserved mechanism of modulation by diacylglycerol for a subset of C. elegans and vertebrate KCNQ/KQT channels, which is dependent upon the carboxyl-terminal domains of channel subunits and activated protein kinase C. PMID- 15797865 TI - ApoA-I lipidation in primary mouse hepatocytes. Separate controls for phospholipid and cholesterol transfers. AB - The liver is the major site of both apolipoprotein A-I (apoA-I) synthesis and ATP binding cassette transporter A1 (ABCA1) expression. Here, we compare the lipidation with cholesterol and phospholipid of newly synthesized human apoA-I (hapoA-I) using adenoviral vector-mediated endogenous expression or exogenously added hapoA-I in wild type and ABCA1-null hepatocytes. Hepatocytes were labeled with [3H]cholesterol (delivered with LDL or methyl-beta-cyclodextrin), [3H]mevalonate, or [3H]choline. ABCA1 deficiency decreased apoA-I phospholipidation by 80%, but acquisition of de novo synthesized and exogenous cholesterol only decreased by 40-60%. The transfer of de novo synthesized cholesterol to apoA-I was decreased at all time points, but that of exogenously delivered cholesterol was independent of ABCA1 activity at the early time points. Progesterone does not affect apoA-I synthesis or its lipidation but inhibited the early phase of apoA-I cholesterol lipidation in both wild type and ABCA1-null hepatocytes. Fast protein liquid chromatography analysis of medium lipoproteins confirmed that with ABCA1 deficiency, the proportion of secreted high density lipoprotein-associated apoA-I and cholesterol decreased by about 50%. The very low density lipoprotein (VLDL)/LDL size fraction also contained a significant level of cholesterol in ABCA1 deficiency, consistent with the result of immunoprecipitations showing the presence of lipoproteins with both apoA-I and murine apoB. ApoA-I lipidation with newly synthesized cholesterol in ABCA1-null hepatocytes was significantly decreased by brefeldin A and monensin. In conclusion, we demonstrate that: (i) whereas most hepatic phospholipidation of apoA-I is mediated by ABCA1, acquisition of cholesterol depends on active transfer from intracellular compartments by ABCA1-dependent and -independent pathways, both sensitive to progesterone and (ii) there is separate regulation of phospholipid and cholesterol lipidation of apoA-I in hepatocytes. PMID- 15797866 TI - Blue light induces mitochondrial DNA damage and free radical production in epithelial cells. AB - Exposure of biological chromophores to ultraviolet radiation can lead to photochemical damage. However, the role of visible light, particularly in the blue region of the spectrum, has been largely ignored. To test the hypothesis that blue light is toxic to non-pigmented epithelial cells, confluent cultures of human primary retinal epithelial cells were exposed to visible light (390-550 nm at 2.8 milliwatts/cm2) for up to 6 h. A small loss of mitochondrial respiratory activity was observed at 6 h compared with dark-maintained cells, and this loss became greater with increasing time. To investigate the mechanism of cell loss, the damage to mitochondrial and nuclear genes was assessed using the quantitative PCR. Light exposure significantly damaged mitochondrial DNA at 3 h (0.7 lesion/10 kb DNA) compared with dark-maintained controls. However, by 6 h of light exposure, the number of lesions was decreased in the surviving cells, indicating DNA repair. Isolated mitochondria exposed to light generated singlet oxygen, superoxide anion, and the hydroxyl radical. Antioxidants confirmed the superoxide anion to be the primary species responsible for the mitochondrial DNA lesions. The effect of lipofuscin, a photoinducible intracellular generator of reactive oxygen intermediates, was investigated for comparison. Exposure of lipofuscin containing cells to visible light caused an increase in both mitochondrial and nuclear DNA lesions compared with non-pigmented cells. We conclude that visible light can cause cell dysfunction through the action of reactive oxygen species on DNA and that this may contribute to cellular aging, age-related pathologies, and tumorigenesis. PMID- 15797867 TI - Use of dominant-negative HrpA mutants to dissect Hrp pilus assembly and type III secretion in Pseudomonas syringae pv. tomato. AB - The Hrp pilus plays an essential role in the long-distance type III translocation of effector proteins from bacteria into plant cells. HrpA is the structural subunit of the Hrp pilus in Pseudomonas syringae pv. tomato (Pst) DC3000. Little is known about the molecular features in the HrpA protein for pilus assembly or for transporting effector proteins. From previous collections of nonfunctional HrpA derivatives that carry random pentapeptide insertions or single amino acid mutations, we identified several dominant-negative mutants that blocked the ability of wild-type Pst DC3000 to elicit host responses. The dominant-negative phenotype was correlated with the disappearance of the Hrp pilus in culture and inhibition of wild-type HrpA protein self-assembly in vitro. Dominant-negative HrpA mutants can be grouped into two functional classes: one class exerted a strong dominant-negative effect on the secretion of effector proteins AvrPto and HopPtoM in culture, and the other did not. The two classes of mutant HrpA proteins carry pentapeptide insertions in discrete regions, which are interrupted by insertions without a dominant-negative effect. These results enable prediction of possible subunit-subunit interaction sites in the assembly of the Hrp pilus and suggest the usefulness of dominant-negative mutants in dissection of the role of the wild-type HrpA protein in various stages of type III translocation: protein exit across the bacterial cell wall, the assembly and/or stabilization of the Hrp pilus in the extracellular space, and Hrp pilus-mediated long-distance transport beyond the bacterial cell wall. PMID- 15797868 TI - Neuroprotective effects of preconditioning ischemia on ischemic brain injury through down-regulating activation of JNK1/2 via N-methyl-D-aspartate receptor mediated Akt1 activation. AB - Our previous studies have demonstrated that the JNK signaling pathway plays an important role in ischemic brain injury and is mediated via glutamate receptor 6. Others studies have shown that N-methyl-d-aspartate (NMDA) receptor is involved in the neuroprotection of ischemic preconditioning. Here we examined whether ischemic preconditioning down-regulates activation of the mixed lineage kinase JNK signaling pathway via NMDA receptor-mediated Akt1 activation. In our present results, ischemic preconditioning could not only inhibit activations of mixed lineage kinase 3, JNK1/2, and c-Jun but also enhanced activation of Akt1. In addition, both NMDA (an agonist of NMDA receptor) and preconditioning showed neuroprotective effects. In contrast, ketamine, an antagonist of NMDA receptor, prevented the above effects of preconditioning. Further studies indicated that LY294002, an inhibitor of phosphoinositide 3-kinase that is an upstream signaling protein of Akt1, could block neuroprotection of preconditioning, and KN62, an inhibitor of calmodulin-dependent protein kinase, also achieved the same effects as LY294002. Therefore, both phosphoinositide 3-kinase and calmodulin-dependent protein kinase are involved in the activation of Akt1 in ischemic tolerance. Taken together, our results indicate that preconditioning can inhibit activation of JNK signaling pathway via NMDA receptor-mediated Akt1 activation and induce neuroprotection in hippocampal CA1 region. PMID- 15797870 TI - A novel isoform of SK2 assembles with other SK subunits in mouse brain. AB - The SK2 subtype of small conductance Ca2+-activated K+ channels is widely distributed throughout the central nervous system and modulates neuronal excitability by contributing to the afterhyperpolarization that follows an action potential. Western blots of brain membrane proteins prepared from wild type and SK2-null mice reveal two isoforms of SK2, a 49-kDa band corresponding to the previously reported SK2 protein (SK2-S) and a novel 78-kDa form. Complementary DNA clones from brain and Western blots probed with an antibody specific for the longer form, SK2-L, identified the larger molecular weight isoform as an N terminally extended SK2 protein. The N-terminal extension of SK2-L is cysteine rich and mediates disulfide bond formation between SK2-L subunits or with heterologous proteins. Immunohistochemistry revealed that in brain SK2-L and SK2 S are expressed in similar but not identical patterns. Heterologous expression of SK2-L results in functional homomeric channels with Ca2+ sensitivity similar to that of SK2-S, consistent with their shared core and intracellular C-terminal domains. In contrast to the diffuse, uniform surface distribution of SK2-S, SK2-L channels cluster into sharply defined, distinct puncta suggesting that the extended cysteine-rich N-terminal domain mediates this process. Immunoprecipitations from transfected cells and mouse brain demonstrate that SK2 L co-assembles with the other SK subunits. Taken together, the results show that the SK2 gene encodes two subunit proteins and suggest that native SK2-L subunits may preferentially partition into heteromeric channel complexes with other SK subunits. PMID- 15797869 TI - Akt-dependent expression of NAIP-1 protects neurons against amyloid-{beta} toxicity. AB - Neurotrophins are a family of growth factors that attenuate several forms of pathological neuronal cell death and may represent a putative therapeutic approach to neurodegenerative diseases. In Alzheimer disease, amyloid-beta (Abeta) is thought to play a central role in the neuronal death occurring in brains of patients. In the present study, we evaluate the ability of neurotrophin 3 (NT-3) to protect neurons against the toxicity induced by aggregated Abeta. We showed that in primary cultures of cortical neurons, NT-3 reduces Abeta-induced apoptosis by limiting caspase-8, caspase-9, and caspase-3 cleavage. This neuroprotective effect of NT-3 was concomitant to an increased level of Akt phosphorylation and was abolished by an inhibitor of the phosphatidylinositol-3 kinase (PI-3K), LY294002. In parallel, NT-3 treatment reduced Abeta induced caspase-3 processing to control levels. In an attempt to link PI-3K/Akt to caspase inhibition, we evaluated the influence of the PI-3K/Akt axis on the expression of a member of the inhibitors of apoptosis proteins (IAPs), the neuronal apoptosis inhibitory protein-1. We demonstrated that NT-3 induces an up regulation of neuronal apoptosis inhibitory protein-1 expression in neurons that promotes the inhibition of Abeta-induced neuronal apoptosis. Together, these findings demonstrate that NT-3 signaling counters Abeta-dependent neuronal cell death and may represent an innovative therapeutic intervention to limit neuronal death in Alzheimer disease. PMID- 15797871 TI - Increased very low density lipoprotein secretion and gonadal fat mass in mice overexpressing liver DGAT1. AB - Acyl-CoA:diacylglycerol acyltransferases (DGATs) catalyze the last step in triglyceride (TG) synthesis. The genes for two DGAT enzymes, DGAT1 and DGAT2, have been identified. To examine the roles of liver DGAT1 and DGAT2 in TG synthesis and very low density lipoprotein (VLDL) secretion, liver DGAT1- and DGAT2-overexpressing mice were created by adenovirus-mediated gene transfection. DGAT1-overexpressing mice had markedly increased DGAT activity in the presence of the permeabilizing agent alamethicin. This suggests that DGAT1 possesses latent DGAT activity on the lumen of the endoplasmic reticulum. DGAT1-overexpressing mice showed increased VLDL secretion, resulting in increased gonadal (epididymal or parametrial) fat mass but not subcutaneous fat mass. The VLDL-mediated increase in gonadal fat mass might be due to the 4-fold greater expression of the VLDL receptor protein in gonadal fat than in subcutaneous fat. DGAT2 overexpressing mice had increased liver TG content, but VLDL secretion was not affected. These results indicate that DGAT1 but not DGAT2 has a role in VLDL synthesis and that increased plasma VLDL concentrations may promote obesity, whereas increased DGAT2 activity has a role in steatosis. PMID- 15797872 TI - CO-dependent activity-controlling mechanism of heme-containing CO-sensor protein, neuronal PAS domain protein 2. AB - Neuronal PAS domain protein 2, which was recently established to be a heme protein, acts as a CO-dependent transcription factor. The protein consists of the basic helix-loop-helix domain and two heme-containing PAS domains (PAS-A and PAS B). In this study, we prepared wild type and mutants of the isolated PAS-A domain and measured resonance Raman spectra of these proteins. Upon excitation of the Raman spectrum at 363.8 nm, a band assignable to Fe3+-S stretching was observed at 334 cm(-1) for the ferric wild type protein; in contrast, this band was drastically weaker in the spectrum of C170A, suggesting that Cys170 is an axial ligand of the ferric heme. The Raman spectrum of the reduced form of wild type was mainly of six-coordinate low spin, and the nu11 band, which is sensitive to the donor strength of the axial ligand, was lower than that of reduced cytochrome c3, suggesting coordination of a strong ligand and thus a deprotonated His. In the reduced forms of H119A and H171A, the five-coordinate species became more prevalent, whereas no such changes were observed for C170A, indicating that His119 and His171, but not Cys170, are axial ligands in the ferrous heme. This means that ligand replacement from Cys to His occurs upon heme reduction. The nu(Fe-CO) versus nu(C-O) correlation indicates that a neutral His is a trans ligand of CO. Our results support a mechanism in which CO binding disrupts the hydrogen bonding of His171 with surrounding amino acids, which induces conformational changes in the His171-Cys170 moiety, leading to physiological signaling. PMID- 15797873 TI - Egg case protein-1. A new class of silk proteins with fibroin-like properties from the spider Latrodectus hesperus. AB - Spiders produce multiple types of silk that exhibit diverse mechanical properties and biological functions. Most molecular studies of spider silk have focused on fibroins from dragline silk and capture silk, two important silk types involved in the survival of the spider. In our studies we have focused on the characterization of egg case silk, a third silk fiber produced by the black widow spider, Latrodectus hesperus. Analysis of the physical structure of egg case silk using scanning electron microscopy demonstrates the presence of small and large diameter fibers. By using the strong protein denaturant 8 M guanidine hydrochloride to solubilize the fibers, we demonstrated by SDS-PAGE and protein silver staining that an abundant component of egg case silk is a 100-kDa protein doublet. Combining matrix-assisted laser desorption ionization tandem time-of flight mass spectrometry and reverse genetics, we have isolated a novel gene called ecp-1, which encodes for one of the protein components of the 100-kDa species. BLAST searches of the NCBInr protein data base using the primary sequence of ECP-1 revealed similarity to fibroins from spiders and silkworms, which mapped to two distinct regions within the ECP-1. These regions contained the conserved repetitive fibroin motifs poly(Ala) and poly(Gly-Ala), but surprisingly, no larger ensemble repeats could be identified within the primary sequence of ECP-1. Consistent with silk gland-restricted patterns of expression for fibroins, ECP-1 was demonstrated to be predominantly produced in the tubuliform gland, with lower levels detected in the major and minor ampullate glands. ECP-1 monomeric units were also shown to assemble into higher aggregate structures through the formation of disulfide bonds via a unique cysteine-rich N terminal region. Collectively, our findings provide new insight into the components of egg case silk and identify a new class of silk proteins with distinctive molecular features relative to traditional members of the spider silk gene family. PMID- 15797874 TI - NF-kappaB and not the MAPK signaling pathway regulates GADD45beta expression during acute inflammation. AB - The GADD45 (growth arrest and DNA damage-inducible) family of genes is involved in the regulation of cell cycle progression and apoptosis. To study signaling pathways affecting GADD45beta expression and to examine systematically in vivo the GADD45beta expression in tissues following various toxic stresses, we created a transgenic mouse by fusing the GADD45beta promoter to firefly luciferase (Gadd45beta-luc). In vivo GADD45beta expression was assessed by measuring the luciferase activity in the Gadd45beta-luc transgenic mouse using a non-invasive imaging system (IVIS Imaging System, Xenogen Corporation). We found that a number of agents that induce oxidative stress, such as sodium arsenite, CCl4, lipopolysaccharide (LPS), or tumor necrosis factor-alpha, are able to induce luciferase expression throughout the entire animal. In liver, spleen, lung, intestine, kidney, and heart, we observed an induction of luciferase activity after LPS treatment, which correlates with an increase of GADD45beta mRNA in these tissues. Processes that induce DNA damage activate the NF-kappaB signaling pathway. Several inhibitors of the NF-kappaB signaling pathway, including dexamethasone, thalidomide, and a proteasome inhibitor, bortezomib, showed inhibitory effects on LPS-induced GADD45beta expression as indicated by a decrease of the luciferase activity. Northern blot analysis confirmed a broad inhibitory effect of bortezomib on LPS-induced GADD45beta mRNA expression in spleen, lung, and intestine. In liver of bortezomib-treated mice, we observed a reverse correlation between the luciferase activity and the GADD45beta mRNA level. We speculate that such a discrepancy could be due to severe liver toxicity caused by bortezomib and LPS co-treatment. MAPK inhibitors had transient and inconsistent effects on LPS-induced luciferase expression. Our data are consistent with the notion that NF-kappaB, but not the MAPK signaling pathways, is involved in the in vivo regulation of GADD45beta expression. Thus, NF-kappaB signaling involves induction of GADD45beta expression, which supports the proposed role of GADD45beta in protecting cells against DNA damaged under various stress conditions. PMID- 15797875 TI - Dissection of synapse induction by neuroligins: effect of a neuroligin mutation associated with autism. AB - To study synapse formation by neuroligins, we co-cultured hippocampal neurons with COS cells expressing wild type and mutant neuroligins. The large size of COS cells makes it possible to test the effect of neuroligins presented over an extended surface area. We found that a uniform lawn of wild type neuroligins displayed on the cell surface triggers the formation of hundreds of uniformly sized, individual synaptic contacts that are labeled with neurexin antibodies. Electron microscopy revealed that these artificial synapses contain a presynaptic active zone with docked vesicles and often feature a postsynaptic density. Neuroligins 1, 2, and 3 were active in this assay. Mutations in two surface loops of neuroligin 1 abolished neuroligin binding to neurexin 1beta, a presumptive presynaptic binding partner for postsynaptic neuroligins, and blocked synapse formation. An analysis of mutant neuroligins with an amino acid substitution that corresponds to a mutation described in patients with an autistic syndrome confirmed previous reports that these mutant neuroligins have a compromised capacity to be transported to the cell surface. Nevertheless, the small percentage of mutant neuroligins that reached the cell surface still induced synapse formation. Viewed together, our data suggest that neuroligins generally promote artificial synapse formation in a manner that is associated with beta neurexin binding and results in morphologically well differentiated synapses and that a neuroligin mutation found in autism spectrum disorders impairs cell surface transport but does not completely abolish synapse formation activity. PMID- 15797876 TI - Specific IgE to colophony? AB - BACKGROUND: Colophony (rosin) is a natural product obtained from coniferous trees. It is used in a diverse range of products such as adhesives, ink, paints and soldering fluxes. Some workers exposed to colophony during soldering can develop occupational asthma; at present, no specific IgE test is available to assess sensitization to colophony. METHODS: Serum samples were obtained from exposed symptomatic individuals (n = 7), some with a likely diagnosis of occupational asthma, exposed asymptomatic individuals (n = 10) and unexposed individuals (n = 11). Serum was tested for specific IgE antibodies against a protein extract produced following in vitro challenge of mono-mac-6 cells with colophony extract. RESULTS: Serum from exposed symptomatic individuals showed increased binding of specific IgE antibodies to a range of colophony-cell protein conjugates [29% (2/7) of samples tested when cut-off > 0.1 or 86% (6/7) of samples tested when cut-off > 0%] compared with both the exposed asymptomatic [0% when cut-off > 0.1, or 20% when cut-off > 0% (2/10)] and the non-exposed control populations [0% when cut-off > 0.1, or 27% when cut-off > 0% (3/11)]. CONCLUSIONS: This novel approach for the production of conjugates to assess sensitization to colophony was able to detect specific IgE in colophony-exposed workers with a likely diagnosis of occupational asthma. PMID- 15797877 TI - A survey of the provision of educational supervision in occupational medicine in the Armed Forces. AB - OBJECTIVE: The aim of this study was to assess whether balanced educational supervision was being provided for occupational medicine (OM) specialist registrars (SpRs) in the Armed Forces and to consider the effectiveness of current supervision. METHODS: Anonymized postal questionnaires, using Likert five point response scales, were sent to Service OM SpRs and their educational supervisors (Ed Sup) to assess the degree of understanding of the purpose, effectiveness and organization of the educational supervision being provided. RESULTS: Completed questionnaires were returned from 35 (92%) specialists and 14 (61%) SpRs. Less than a third of the specialists were involved as Ed Sup and only three (11%) had undergone any formal medical education training. There was an agreement about the purpose and effectiveness of educational supervision between both the groups, though there appeared to be a bias towards providing supportive functions with patient management and educational components scoring less highly. CONCLUSIONS: A high response rate enabled an accurate assessment of OM supervision practices in the Armed Forces to be made. There was agreement over the relative effectiveness, importance and coverage of educational supervision, though this was being provided by a relatively small group of OM specialists, few of whom had undertaken any formal medical education training. The supervision provided also appeared to be biased towards providing supportive functions rather than patient management and educational components of training, which may result in an unbalanced training for the SpRs. Having assessed the relatively close-knit service OM community, there would be value in seeing whether similar patterns prevail in the wider OM community. PMID- 15797878 TI - Interleukin-1 gene cluster polymorphisms and alcoholism in Spanish men. AB - AIMS: In an attempt to explain differences in susceptibility to alcoholism and alcohol liver disease (ALD), different genes have been analysed, among them those encoding inflammatory cytokines. Thus, it has been reported recently that both the interleukin 1 receptor antagonist (IL1RN) and the IL1beta (IL1B) genes may influence the risk of ALD in Japanese alcoholics. We analysed the distribution of single nucleotide polymorphisms (SNPs) located in the IL1A, IL1B, IL1R1 and IL1RN genes in alcoholic and non-alcoholic Spanish subjects. METHODS: DNA samples were obtained from 139 male alcoholics, 78 of whom were diagnosed as alcohol dependent (32 patients with liver cirrhosis and 46 without ALD) and 61 as alcohol abusers (25 with liver cirrhosis and 36 without ALD). As a control, we studied 81 age- and sex-matched healthy volunteers. RESULTS: Alleles -511 IL1B*1 and IL1RN*1 were represented more in alcoholic patients than in the control group. We did not find any association of alcoholism or ALD with polymorphisms in the IL1A and IL1R1 genes. CONCLUSIONS: We conclude that the proteins encoded by the IL1RN and IL1B genes may be involved in susceptibility to alcoholism in Spanish men, probably through a different pathway from that involved in the regulation of the inflammatory response. PMID- 15797879 TI - Recent trends in incidence rates for selected alcohol-related cancers in the United States. AB - AIMS: To examine recent trends in incidence rates for cancer types most strongly associated with alcohol use, using data from US cancer registries. METHODS: Age standardized annual incidence rates (ASIRs) for squamous cell carcinomas of the oral cavity and pharynx, esophagus and larynx diagnosed in the most recent 10 year period (1992-2001) were examined for geographic areas included in the US National Cancer Institute's Surveillance, Epidemiology and End Results (SEER) program of high-quality cancer registries. RESULTS: For all geographic areas combined, ASIRs for these cancers declined over time, with no evidence for a recent plateau or upturn. This also held true for ASIRs in younger adults (age 20 54 years at diagnosis). For white males, declines in ASIRs occurred in each of the 11 geographic areas, and were statistically significant in nine areas. The declines in ASIRs were consistent with temporal declines in apparent alcohol consumption by state, although the prevalence of binge and heavy drinking in adults increased in some states. CONCLUSIONS: Although there was no evidence for a recent plateau in ASIRs, continued surveillance is needed, in view of recent increases in the prevalence of binge and heavy drinking among US adults. PMID- 15797880 TI - Hazardous drinking in elite New Zealand sportspeople. AB - AIMS: To examine the link between hazardous drinking and level of sport participation in New Zealand. METHODS: Sports science and general university students (n = 427) completed a sporting profile questionnaire that included the Alcohol Use Disorders Identification Test (AUDIT). RESULTS: Elite sportspeople (both provincial and international/country level) reported higher rates of hazardous drinking than non-sportspeople and non-elite sportspeople. Similar differences were observed in AUDIT subscale scores, with international/country level sportspeople reporting greater symptoms of dependence than other groups. PMID- 15797881 TI - The effect of data collection mode and ethnicity of interviewer on response rates and self-reported alcohol use among Turks and Moroccans in the Netherlands: an experimental study. AB - AIMS: To test the effects of data collection mode and ethnicity of interviewers on response rates and self-reported alcohol use among second-generation Turks and Moroccans in Rotterdam, The Netherlands. METHODS: Two hundred and sixty-nine Turks and 271 Moroccans were interviewed face-to-face, and 475 Turks and 482 Moroccans received a mailed questionnaire. Half of the Turks and Moroccans randomly allocated to the interview mode were ethnically matched to the interviewer; the remainder were allocated to a Dutch interviewer. RESULTS: Turks and Moroccans more often responded to a face-to-face interview than to a mailed questionnaire. No effect of ethnicity of interviewer on response rates was demonstrated. With respect to the effects on alcohol reports, Turks and Moroccans tended to report a higher alcohol use in the mailed survey than in the face-to face interview. They reported significantly more often excessive drinking in the mail survey than in the face-to-face interviews. Ethnicity of the interviewer resulted in Turks and Moroccans reporting a higher prevalence of alcohol use during the previous 6 months when interviewed by a Dutch interviewer compared with an ethnically matched interviewer. CONCLUSIONS: Among second-generation Turks and Moroccans, mail surveys seem most suitable to measure mean and excessive alcohol use. However, interviews held by Dutch interviewers seem to be the most appropriate method to study the prevalence of alcohol use during the previous 6 months. PMID- 15797882 TI - Sex differences in spatial visualization and episodic memory as a function of alcohol consumption. AB - AIMS: Sex differences in visuospatial ability as well as episodic memory have been reliably demonstrated, irrespective of alcoholism. Studies in alcoholics have consistently documented cognitive deficits in visuospatial ability, problem solving and memory function. This cross-sectional, population-based study examined if sex differences in cognitive performance could be impacted by alcohol consumption. METHODS: Drinking data were collected from 2224 randomly sampled adults, aged between 35 and 85 years, who participated in the Betula study on memory, health and aging. Participants were classified into non-, light, moderate and heavy drinking subgroups based on sex-adjusted normative values. Cognitive tasks demonstrating clear sex differences, such as episodic memory tasks (favouring women) and spatial visualization tasks (favouring men), were conducted and performance was assessed by sex and the drinking group. RESULTS: After controlling for age and education, overall analyses found expected sex differences in episodic memory and spatial visualization that were apparent across the entire population. When these sex differences were examined by drinking group, visuospatial performance favouring men disappeared for the moderate to heavy drinking groups, but higher performance by women on episodic memory tasks was consistent across all levels of alcohol consumption. Traditional biomarkers of increased alcohol consumption (GGT and MCV) correlated with the reported drinks/day. CONCLUSIONS: These results lend support to the theory that moderate alcohol intake may be beneficial to cognitive function in women, but not necessarily in men. PMID- 15797883 TI - Screening for hazardous or harmful drinking using one or two quantity-frequency questions. AB - AIMS: To address the accuracy of quantity-frequency (QF) questions in screening for hazardous or harmful drinking. METHODS: Three groups were interviewed: patients presenting to emergency departments for care of an acute injury (n = 1537) or a medical illness (n = 1151), and community controls interviewed by telephone (n = 1112). The first question about alcohol was a single alcohol screening question (SASQ), 'When was the last time you had more than X drinks in one day?', where X = 4 for women and 5 for men, with any time in the past 3 months considered a positive screen (1 drink = 14 g ethanol). The subsequent alcohol questions were a calendar-based review of recent drinking and the alcohol questions from the diagnostic interview schedule (DIS), which included questions about usual frequency and average quantity. Hazardous drinking was defined as drinking >4 drinks in 1 day or >14 drinks in 1 week for men (women 3 and 7) (Guidelines of the US National Institute on Alcohol Abuse and Alcoholism). Current alcohol use disorders were defined using DSM-IV criteria. The areas under the receiver operating characteristic (ROC) curves in identifying hazardous drinking or current alcohol use disorder were compared. RESULTS: The area under the ROC curves in the three samples combined were 0.81 for SASQ (95% confidence interval (CI) 0.79-0.82), 0.80 for a question about average quantity alone (0.79 0.82) and 0.85 for the product of usual frequency times average quantity (0.84 0.86). The QF product and the question about average quantity performed consistently across the three groups. CONCLUSIONS: In clinical settings, one way to put these findings into practice is to screen first with a single question, such as the SASQ, a single question about typical quantity, or a question about the frequency of heavy drinking such as the third item of the alcohol use disorders test (AUDIT). PMID- 15797884 TI - Comparison of serum fatty acid ethyl esters and urinary 5-hydroxytryptophol as biochemical markers of recent ethanol consumption. AB - AIMS: To examine the effects of an acute dose of ethanol on serum fatty acid ethyl esters (FAEEs) concentration and urinary 5-hydroxytryptophol (5-HTOL)/5 hydroxyindole-3-acetic acid (5-HIAA) ratio. METHODS: Sixteen (14 male, 2 female) heavy alcohol drinkers were tested in a single, 2-day long session. Six participants received 1.5 g/l of ethanol/l of body water (approximately 0.75 g/kg of body weight, low dose group: LD) and 10 participants received 2.0 g/l of ethanol ( approximately 1.0 g/kg of body weight, high dose group: HD) in four divided doses every 20 min. Blood, urine, and breath samples were collected repeatedly over 36 h following the ingestion of ethanol and were analyzed for the presence of FAEE, 5-HTOL/5-HIAA, and ethanol, respectively. Serum gamma glutamyltransferase (GGT), a marker of chronic ethanol use, was also included. RESULTS: The breath ethanol level peaked approximately 1 h after the last dose, at 95 and 120 mg/dl for the LD and HD groups, respectively. The mean ratio of urinary 5-HTOL/5-HIAA was significantly elevated 5 and 9 h after ethanol administration, but returned to baseline 13 h after ethanol administration. This ratio was twice as high for the HD group compared with the LD group. Serum levels of FAEEs were significantly elevated at 5 h, but not 13 h after ethanol administration. There were no time-dependent changes in serum GGT levels. CONCLUSIONS: Measuring the levels of FAEE and 5-HTOL/5-HIAA ratio provides a convenient method to detect recent, particularly binge-type, ethanol use, but these measures may have limited applicability in detecting ethanol use in traditional clinical trial settings. PMID- 15797885 TI - Clinical predictors of response to naltrexone in alcoholic patients: who benefits most from treatment with naltrexone? AB - AIMS: To determine the clinically ascertained variables that are related to satisfactory response to naltrexone (NTX) treatment of alcohol dependence after detoxification. METHODS: The use of intake and outcome variables were measured in a randomized 3-month open-controlled trial comparing the effects of naltrexone plus psychotherapy treatment versus psychotherapy treatment alone on the maintenance of abstinence in the final 28 days (n = 336, all male). RESULTS: Predictors of a positive response to NTX treatment were family history of alcoholism (P = 0.010), early age at onset of drinking problems (P = 0.014) and comorbid use of other drugs of abuse (P < 0.001). Among the subjects not treated with NTX, the greater the number of predictor variables, the lower the final 28 days abstinence rates (P = 0.00003), but this was not the case in patients treated with NTX (P = 0.844). CONCLUSIONS: Patients with these features, suggesting biological vulnerability overall have poorer outcomes, but this can be reduced with NTX treatment. The type of alcoholism should be considered before deciding on the pharmacological strategy. PMID- 15797886 TI - A challenge to the metabolic approach to myocardial ischaemia. AB - The negative results of glucose-insulin-potassium (GIK) in the very large CREATE ECLA trial that studied 20,201 patients with ST-elevation acute myocardial infarction (AMI), are disappointing and warrant thorough evaluation. We attempt to put the new data into perspective and uncover the serious flaws in the trial design, otherwise the whole metabolic concept will be disparaged. The crucial issue, developed from basic science data, is that GIK should be initiated very early, before, or at the time of reperfusion. Another problem with CREATE-ECLA is that the mortality in Killip class 1 reperfused patients was 7.1%, much higher than that of a recent Dutch study in which mortality was only 1.2%. Nonetheless, there was a strong trend towards a lower mortality in the sub-groups that received the best reperfusion therapy in CREATE-ECLA, as well as in the first of two rather small Dutch GIK trials. In the future, the ideal protocol to test would be if GIK were given in the ambulance as the patient is being transported to a specialized centre of percutaneous coronary intervention (PCI), with the aim of expanding the time window between pain onset and actual PCI. PMID- 15797887 TI - Efficacy of sustained low-efficiency dialysis in the treatment of salicylate toxicity. PMID- 15797888 TI - Unilateral pleural effusions associated with stenoses of left brachiocephalic veins in haemodialysis patients. PMID- 15797889 TI - A single bolus of a low molecular weight heparin to patients on haemodialysis depletes lipoprotein lipase stores and retards triglyceride clearing. AB - BACKGROUND: Low molecular weight heparins (LMWH) are increasingly used during haemodialysis (HD) to prevent clotting in the extracorporeal devices. It has been suggested that LMWH release endothelial-bound lipoprotein lipase (LPL) less efficiently than unfractionated heparin (UFH) does and thereby cause less disturbance of lipid metabolism. Evidence from in vitro studies and from animal experiments indicate, however, that both types of heparin preparations have the same ability to release endothelial LPL, but LMWH are less effective in preventing uptake and degradation of LPL in the liver. Model studies in humans indicate that LMWH cause as much depletion of LPL stores and impaired lipolysis of triglyceride (TG)-rich lipoproteins as UFH does. METHODS: Two anticoagulant regimes based on present clinical practice were compared in nine HD patients. UFH was administered as a primed infusion, whereas the LMWH (dalteparin) was given only as a single bolus pre-dialysis. Blood was sampled regularly for LPL activity and TG. RESULTS: LPL activity in blood was significantly lower during the dialysis with dalteparin. To explore the remaining activity at the endothelium, a bolus of UFH was given after 3 h of dialysis. The bolus brought out about the same amount of LPL, regardless of whether UFH or dalteparin had been used during dialysis. The increase in TG was significantly higher during dialysis with dalteparin. CONCLUSIONS: This study indicates that a single bolus of dalteparin pre-dialysis interferes with the LPL system as much as, or more than an infusion of UFH does. PMID- 15797890 TI - Recurrent IgA nephropathy after renal transplantation despite immunosuppressive regimens with mycophenolate mofetil. AB - BACKGROUND: Transplantation offers an excellent option for patients with immunoglobulin-A nephropathy (IgAN) with severe renal dysfunction. However, IgAN frequently recurs in allografts treated with azathioprine. We examined the impact of mycophenolate mofetil immunosuppression on recurrence of IgAN. METHODS: We reviewed the charts of patients transplanted for IgAN at our institution in the cyclosporin era. Patients were excluded from further analysis if follow-up was <12 months or if immunosuppression at engraftment did not include azathioprine or mycophenolate mofetil. Laboratory data, medications and allograft biopsy findings were compiled. RESULTS: 152 kidney transplantations met the study criteria. At engraftment, 61 allografts were treated with azathioprine and 91 with mycophenolate mofetil. By 3 years post-transplant, IgAN developed in six of 60 (10.0%) azathioprine-treated allografts and five of 62 (8.1%) mycophenolate mofetil-treated allografts (P = 0.76). Overall, 13 azathioprine-treated and seven mycophenolate mofetil-treated allografts showed recurrence. As expected in this retrospective study, the duration of observation was longer in the azathioprine group. The interval between engraftment and diagnosis of recurrent disease was also longer. Survival of allografts with recurrent IgAN was similar in the two groups. Survival of allografts with recurrent IgAN was worse than for allografts without recurrence or allografts transplanted into patients with non-IgAN renal failure. Neither switching azathioprine to mycophenolate mofetil nor using an angiotensin-converting enzyme inhibitor or angiotensin-II type 1 receptor blocker ameliorated the clinical course after a biopsy documented recurrent IgAN. CONCLUSIONS: Mycophenolate mofetil, compared with azathioprine, did not lessen the recurrence of IgAN or its clinical impact. PMID- 15797891 TI - Agonist of peroxisome proliferator-activated receptor-gamma, rosiglitazone, reduces renal injury and dysfunction in a murine sepsis model. AB - BACKGROUND: Agonists of the peroxisome proliferator-activated receptor-gamma may help to regulate inflammation by modulating the production of inflammatory mediators and adhesion molecules. The purpose of this study was to determine the protective effects of rosiglitazone on renal injury in a sepsis model and to explore the mechanism. METHODS: In lipopolysaccharide (LPS)-induced mouse sepsis, we examined the effect of rosiglitazone on LPS-induced overproduction of inflammatory mediators, on the expression of adhesion molecules in renal tubular epithelial cells and on renal function. The mechanism of the protective effect was investigated in vitro using human renal tubular epithelial cells. RESULTS: Rosiglitazone significantly decreased serum tumour necrosis factor (TNF)-alpha and interleukin (IL)-1beta levels during sepsis. The levels of blood urea nitrogen and creatinine were significantly lower in mice pre-treated with rosiglitazone than that in LPS-treated mice. Rosiglitazone reduced the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) in tubular epithelial cells and interstitium of LPS-treated mice. Pre treatment with rosiglitazone reduced the infiltration of macrophages/monocytes in renal tissue. In cultured tubular epithelial cells, rosiglitazone significantly decreased the expression of ICAM-1 and VCAM-1 induced by TNF-alpha or IL-1beta, inhibited the degradation of inhibitor kappaBalpha (IkappaBalpha) and blocked the activation of the p65 subunit of nuclear factor (NF)-kappaB. CONCLUSIONS: These results indicate that pre-treatment with rosiglitazone attenuated the production of TNF-alpha and IL-1beta and reduced adhesion molecule expression in renal tubular epithelial cells of LPS-treated mice. Rosiglitazone has an anti inflammatory effect in renal tubular epithelial cells through the inhibition of NF-kappaB activation. PMID- 15797892 TI - Do we need to change our administration practice with regard to sodium ferric gluconate complex in glucose? PMID- 15797893 TI - Use of clinical syndromes to target antibiotic prescribing in seriously ill children in malaria endemic area: observational study. AB - OBJECTIVES: To determine how well antibiotic treatment is targeted by simple clinical syndromes and to what extent drug resistance threatens affordable antibiotics. DESIGN: Observational study involving a priori definition of a hierarchy of syndromic indications for antibiotic therapy derived from World Health Organization integrated management of childhood illness and inpatient guidelines and application of these rules to a prospectively collected dataset. SETTING: Kilifi District Hospital, Kenya. PARTICIPANTS: 11,847 acute paediatric admissions. MAIN OUTCOME MEASURES: Presence of invasive bacterial infection (bacteraemia or meningitis) or Plasmodium falciparum parasitaemia; antimicrobial sensitivities of isolated bacteria. RESULTS: 6254 (53%) admissions met criteria for syndromes requiring antibiotics (sick young infants; meningitis/encephalopathy; severe malnutrition; very severe, severe, or mild pneumonia; skin or soft tissue infection): 672 (11%) had an invasive bacterial infection (80% of all invasive bacterial infections identified), and 753 (12%) died (93% of all inpatient deaths). Among P falciparum infected children with a syndromic indication for parenteral antibiotics, an invasive bacterial infection was detected in 4.0-8.8%. For the syndrome of meningitis/encephalopathy, 96/123 (76%) isolates were fully sensitive in vitro to penicillin or chloramphenicol. CONCLUSIONS: Simple clinical syndromes effectively target children admitted with invasive bacterial infection and those at risk of death. Malaria parasitaemia does not justify withholding empirical parenteral antibiotics. Lumbar puncture is critical to the rational use of antibiotics. PMID- 15797895 TI - Occupational exposure visualization comes of age. PMID- 15797894 TI - Alternative metrics for noise exposure among construction workers. AB - Although the exposure-response relationships for noise-induced hearing loss are relatively well established, there is not complete agreement on which metrics of noise exposure best represent risk of hearing damage. In particular, while L(eq), based on a 3 dB exchange rate (ER) is used by most agencies, US OSHA's standard is based on the L(avg), which uses a 5 dB ER. In addition, peak levels of exposure, which are commonly found in some industries, including construction, are believed to increase risk above that predicted by the L(eq). This paper presents an analysis of a large database of noise exposures among construction workers, comparing several noise metrics, and their application to a cohort of construction workers. Metrics examined were the L(avg), L(eq) and L(max), expressing average levels of exposure across an exposure interval. Two novel metrics were derived from these monitored metrics, L(eq)/L(avg) and L(max)/L(eq), as measures of exposure variability and 'peakiness', respectively. A total of 730 workshifts, including data on 361 492 min of exposure to workers in nine trades were examined. Correlations between average metrics (L(eq), L(avg) and L(max)) are generally very high, while the variability metrics are poorly correlated with either average levels, or with each other, indicating that they characterize different aspects of exposure. Alternative models for estimating exposure for the cohort were considered and the use of a task-within-trade specific mean level was adopted. The task-specific estimates of exposure using the various metrics will be applied to the cohort's work history to explore the importance of these alternative metrics in estimating risk of noise-induced damage. PMID- 15797896 TI - Chiropterophily in Sinningieae (Gesneriaceae): Sinningia brasiliensis and Paliavana prasinata are bat-pollinated, but P. sericiflora is not. Not yet? AB - BACKGROUND AND AIMS: Based on the bell shape and greenish colour of the flowers, bat-pollination was suggested for some Sinningieae species (Gesneriaceae). Actually, there are no reports on pollination biology and visitors for these species. This paper reports on pollination biology of Sinningia brasiliensis, Paliavana prasinata and P. sericiflora in south-eastern Brazil. METHODS: Flowers were observed in situ to determine phases of anthesis, colour patterns and scent intensity. Corolla measures were taken from fresh flowers. Nectar production and concentration were measured in flowers bagged at the pre-anthesis phases. Direct visual observations of visitors were made during the day and night, and photographs were taken to analyse their visiting behaviour. KEY RESULTS: Some floral features of the three species fit the bat-pollination syndrome: large, robust and gullet-shaped corollas, colour patterns and large amount of nectar. However, other floral features-such as onset of anthesis, nectar attributes and odour intensity-differ among the species. Nectar volume and total sugar production increased significantly at midnight in S. brasiliensis and P. prasinata, but in P. sericiflora there were no significant differences in the total nectar and sugar production during anthesis. Scent intensity is much higher in S. brasiliensis and P. prasinata than P. sericiflora. Flowers of S. brasiliensis and P. prasinata, whose features fit the chiropterophilous syndrome, are pollinated by glossophagine bat species. Paliavana sericiflora, on the other hand, presents floral features intermediate between bat and hummingbird syndromes, but is visited and pollinated only by hummingbirds. CONCLUSIONS: These data strengthen the statement that the bat syndrome in Sinningieae originated independently in Sinningia brasiliensis and in Paliavana species. Paliavana sericiflora may be another example of a plant 'in transition' from the hummingbird to the bat pollination, but a reversion in the direction of bat to hummingbird might not be excluded. It is also possible that this is a case of speciation on both bat and hummingbird pollination. Studies on Paliavana sister species may provide insights about origins and the evolutionary directions of the pollination systems of these species. PMID- 15797897 TI - Polar paths of diffusion across plant cuticles: new evidence for an old hypothesis. AB - BACKGROUND: The plant cuticle is an extracellular lipophilic biopolymer covering leaf and fruit surfaces. Its main function is the protection of land-living plants from uncontrolled water loss. In the past, the permeability of the cuticle to water and to non-ionic lipophilic molecules (pesticides, herbicides and other xenobiotics) was studied intensively, whereas cuticular penetration of polar ionic compounds was rarely investigated. RECENT PROGRESS: Recent work measuring cuticular penetration of inorganic and organic ions is presented; the effects of molecular size of ions, temperature, wax extraction, humidity and plasticizers strongly support the conclusion that ions penetrate cuticles via water-filled pores. The cuticle covering stomata and trichomes forms the preferential site of ion penetration. This indicates that cuticles possess a pronounced lateral heterogeneity: the largest fraction of the cuticle surface is covered by the lipophilic domains of cutin and wax, but to a certain extent polar domains are also present in the cuticle, which form preferential sites of penetration for polar compounds. THE FUTURE: The chemical nature of these polar domains awaits detailed characterization, which will be of major importance in agriculture and green biotechnology, since polar paths of diffusion represent the most important transport routes for foliar-applied nutrients. Furthermore, many compounds acting as inducers of gene expression in transgenic plants are ionic and need to penetrate the cuticle via polar paths in order to be active. PMID- 15797898 TI - Comparative structure and pollen production of the stamens and pollinator deceptive staminodes of Commelina coelestis and C. dianthifolia (Commelinaceae). AB - BACKGROUND AND AIMS: Flowers of Commelina coelestis and C. dianthifolia provide pollen alone as a floral reward, and rely on visual cues to attract pollinators. Three stamen types, all producing pollen, occur in each of these species: two cryptically coloured lateral stamens, a single cryptically coloured central stamen and three bright yellow staminodes that sharply contrast with the blue to purple corolla. The objective was to compare the stamen structure and pollen characteristics of each of the three stamen types, and to test the hypothesis that the staminodes are poor contributors of viable pollen for the siring of seed. The pollination roles of the three stamen types and the breeding systems of both species were also explored. METHODS: Light, fluorescence and scanning electron microscopy were utilized to examine stamen morphology and pollen structure and viability. Controlled hand pollinations were used to explore the breeding system of each species. Filament and style lengths were measured to investigate herkogamy and autogamy. KEY RESULTS: Pollen from all stamen morphs is viable, but staminode pollen has significantly lower viability. Pollen polymorphism exists both (a) between the lateral and central stamens and the staminodes, and (b) within each anther. Lateral and central stamens have thicker endothecia with a greater number of secondary cell wall thickenings than the staminodes. CONCLUSIONS: Both species are entomophilous and facultatively autogamous. Lateral stamen pollen is important for cross-pollination, central stamen pollen is utilized by both species as a pollinator reward and for delayed autogamy in C. dianthifolia, and the staminodes mimic, by means of both colour and epidermal features, large amounts of pollen to attract insects to the flowers. Pollen from all three anther morphs is capable of siring seed, although staminode pollen is inferior. The thin staminode endothecium with fewer secondary thickenings retards staminode dehiscence. PMID- 15797899 TI - Herkogamy and mating patterns in the self-compatible daffodil Narcissus longispathus. AB - BACKGROUND AND AIMS: Floral design in self-compatible plants can influence mating patterns. This study investigated Narcissus longispathus, a self-compatible bee pollinated species with wide variation in anther-stigma separation (herkogamy), to determine the relationship between variation in this floral trait and the relative amounts of cross- and self-fertilization. METHODS: Anther-stigma separation was measured in the field in six populations of N. longispathus from south-eastern Spain. Variation in herkogamy during the life of individual flowers was also quantified. Multilocus outcrossing rates were estimated from plants differing in herkogamy using allozyme markers. KEY RESULTS: Anther-stigma separation varied considerably among flowers within the six populations studied (range = 1-10 mm). This variation was nearly one order of magnitude larger than the slight, statistically non-significant developmental variation during the lifespan of individual flowers. Estimates of multilocus outcrossing rate for different herkogamy classes (t(m) range = 0.49-0.76) failed to reveal a monotonic increase with increasing herkogamy. CONCLUSIONS: It is suggested that the lack of a positive relationship between herkogamy and outcrossing rate, a result that has not been previously documented for other species, could be mostly related to details of the foraging behaviour of pollinators. PMID- 15797900 TI - Evaluating an internet weight loss program for diabetes prevention. AB - Being overweight or obese is a major risk factor for developing type 2 diabetes but weight loss through lifestyle interventions can markedly reduce its incidence. The Internet provides an opportunity for the development and implementation of lifestyle intervention programs that promote self-managed behavioural change. We developed an online weight loss program emphasizing physical activity and dietary modifications and conducted a short-term qualitative evaluation of it, examining participant recruitment from the general public, website usage and satisfaction and use of self-reported health risk appraisal records. From a total of 808 registered participants who accessed the online services, 683 (84.5%) completed at least one online health risk appraisal and of those, 464 (68%) people (364 female, aged 19-70 years; 100 male, aged 20 71 years) enrolled in the weight loss program. The program was met with a high level of satisfaction by participants, with 56% of feedback respondents agreeing that the program helped them achieve their goals. The program home page, the principal arrival destination of participants, was viewed an average of 29 times per participant, suggesting that the website's services were used with a high frequency. These preliminary findings indicate that the general public will use an Internet-based weight loss program that involves physical activity and dietary behavioural interventions. Whether Internet delivery of these interventions can significantly reduce the risk for developing type 2 diabetes is worthy of further investigation. The findings have relevance for development of health promotion policies and practices. PMID- 15797901 TI - Rural women's knowledge of AIDS in the higher prevalence states of India: reproductive health and sociocultural correlates. AB - This study aimed to identify socio-cultural and reproductive health correlates of knowledge about AIDS among rural women using multivariate analysis of 1998-1999 National Family Health Survey (NFHS) data from two Indian states, Maharashtra and Tamil Nadu, where the urban HIV prevalence is relatively high. Analysis using multiple logistic regression was undertaken, modelling women's knowledge of AIDS, of whether the disease can be avoided, and of effective means of protection. Although 47% of all rural women in Maharashtra were aware of AIDS only about 28% knew that one can avoid it, and only about 16% possessed correct knowledge about its transmission. In Tamil Nadu, where overall 82% of rural women had awareness of AIDS, about 71% knew that one can avoid the disease but only about 31% possessed correct knowledge about its transmission. In both states, women from socially and economically backward groups had lower odds both of having awareness of AIDS and knowledge of ways to avoid getting the disease. Associations with socio-cultural and reproductive variables and the impact of contact with family planning services differed in the two states. The spread of the epidemic to rural areas presents a need actively to disseminate AIDS related knowledge for health protection rather than waiting for knowledge to follow the appearance of the disease in communities. Approaches to health promotion that do not consider differing contextual factors are unlikely to succeed. In particular, innovative strategies to disseminate knowledge among disadvantaged population groups are needed. PMID- 15797902 TI - Development of a collaborative model to improve school health promotion in The Netherlands. AB - In recent decades, school health promotion programs have been developing into whole-school health approaches. This has been accompanied by a greater understanding among health promoters of the core-business of schools, namely education, and how health promotion objectives can be integrated into this task. Evidence of the positive impact of school health promotion on health risk behavior of students is increasing. This article focuses on the processes and initial results of developing a collaborative model tailored for whole-school health in the Netherlands, named schoolBeat. The Dutch situation is characterized by fragmentation, a variety of health and welfare groups supporting schools, and a lack of sound integrated youth policies. A literature review, observations, and stakeholder consultation provided a clear picture of the current situation in school health promotion, and factors limiting a comprehensive and needs-based approach to school health. This revealed that a health promotion team within a school is fundamental to an effective approach to tailored school health promotion. A respected member of school staff should chair this team. To strengthen the link with the school care team, the school care coordinator should be a member of both teams. To provide coordinated support to all schools in a region, participating organizations decided to share advisory tasks. These tasks are included in the regular health promotion work of their staff. This means working with one advisor representing all school-health organizations per school, and using a comprehensive overview of possible support and projects promoting health. Empowering schools in needs assessments and comprehensive school health promotion is an important element of the developed approach. This article concludes with an examination of emerging issues in evaluating collaborative school health support during the first 18 months of development, and implementation and future perspectives regarding sustainable collaboration and quality improvement. PMID- 15797903 TI - Mining SARS-CoV protease cleavage data using non-orthogonal decision trees: a novel method for decisive template selection. AB - MOTIVATION: Although the outbreak of the severe acute respiratory syndrome (SARS) is currently over, it is expected that it will return to attack human beings. A critical challenge to scientists from various disciplines worldwide is to study the specificity of cleavage activity of SARS-related coronavirus (SARS-CoV) and use the knowledge obtained from the study for effective inhibitor design to fight the disease. The most commonly used inductive programming methods for knowledge discovery from data assume that the elements of input patterns are orthogonal to each other. Suppose a sub-sequence is denoted as P2-P1-P1'-P2', the conventional inductive programming method may result in a rule like 'if P1 = Q, then the sub sequence is cleaved, otherwise non-cleaved'. If the site P1 is not orthogonal to the others (for instance, P2, P1' and P2'), the prediction power of these kind of rules may be limited. Therefore this study is aimed at developing a novel method for constructing non-orthogonal decision trees for mining protease data. RESULT: Eighteen sequences of coronavirus polyprotein were downloaded from NCBI (http://www.ncbi.nlm.nih.gov). Among these sequences, 252 cleavage sites were experimentally determined. These sequences were scanned using a sliding window with size k to generate about 50,000 k-mer sub-sequences (for short, k-mers). The value of k varies from 4 to 12 with a gap of two. The bio-basis function proposed by Thomson et al. is used to transform the k-mers to a high-dimensional numerical space on which an inductive programming method is applied for the purpose of deriving a decision tree for decision-making. The process of this transform is referred to as a bio-mapping. The constructed decision trees select about 10 out of 50,000 k-mers. This small set of selected k-mers is regarded as a set of decisive templates. By doing so, non-orthogonal decision trees are constructed using the selected templates and the prediction accuracy is significantly improved. PMID- 15797904 TI - A bioinformatic screen for novel A-I RNA editing sites reveals recoding editing in BC10. AB - MOTIVATION: Recent studies have demonstrated widespread adenosine-inosine RNA editing in non-coding sequence. However, the extent of editing in coding sequences has remained unknown. For many of the known sites, editing can be observed in multiple species and often occurs in well-conserved sequences. In addition, they often occur within imperfect inverted repeats and in clusters. Here we present a bioinformatic approach to identify novel sites based on these shared features. Mismatches between genomic and expressed sequences were filtered to remove the main sources of false positives, and then prioritized based on these features. This protocol is tailored to identifying specific recoding editing sites, rather than sites in non-coding repeat sequences. RESULTS: Our protocol is more sensitive for identifying known coding editing sites than any previously published mammalian screen. A novel multiply edited transcript, BC10, was identified and experimentally verified. BC10 is highly conserved across a range of metazoa and has been implicated in two forms of cancer. PMID- 15797905 TI - Identification of transcription factor binding sites with variable-order Bayesian networks. AB - MOTIVATION: We propose a new class of variable-order Bayesian network (VOBN) models for the identification of transcription factor binding sites (TFBSs). The proposed models generalize the widely used position weight matrix (PWM) models, Markov models and Bayesian network models. In contrast to these models, where for each position a fixed subset of the remaining positions is used to model dependencies, in VOBN models, these subsets may vary based on the specific nucleotides observed, which are called the context. This flexibility turns out to be of advantage for the classification and analysis of TFBSs, as statistical dependencies between nucleotides in different TFBS positions (not necessarily adjacent) may be taken into account efficiently--in a position-specific and context-specific manner. RESULTS: We apply the VOBN model to a set of 238 experimentally verified sigma-70 binding sites in Escherichia coli. We find that the VOBN model can distinguish these 238 sites from a set of 472 intergenic 'non promoter' sequences with a higher accuracy than fixed-order Markov models or Bayesian trees. We use a replicated stratified-holdout experiment having a fixed true-negative rate of 99.9%. We find that for a foreground inhomogeneous VOBN model of order 1 and a background homogeneous variable-order Markov (VOM) model of order 5, the obtained mean true-positive (TP) rate is 47.56%. In comparison, the best TP rate for the conventional models is 44.39%, obtained from a foreground PWM model and a background 2nd-order Markov model. As the standard deviation of the estimated TP rate is approximately 0.01%, this improvement is highly significant. PMID- 15797906 TI - Formalizing concepts of species, sex and developmental stage in anatomical ontologies. AB - MOTIVATION: Anatomy ontologies have a growing role in bioinformatics-for example, in indexing gene expression data in model organisms. To relate or draw conclusions from data so indexed, anatomy ontologies must be equipped with the formal vocabulary that would allow statements about meronomy to be qualified by constraints such as part of the male or part at the embryonic stage. Lacking such a vocabulary, anatomists have built this information into the structure of the ontology or into anatomical terms. For example, in the FlyBase anatomy for drosophila, the term larval abdominal segment encodes the stage in the term, while the terms male genital disc and female genital disc encode the sex. It remains implicit that a fly has one and only one of these parts during its larval stage. Such indicators of context can and should be represented explicitly in the ontology. RESULTS: The framework we have defined for anatomical ontologies allows the canonical anatomy structures of a given species to be those common to all sexes, and to have either male, female or hermaphrodite parts--but not combinations of the latter. Temporal aspects of development are addressed by associating a stage with organism parts and requiring a connected anatomy to have parts that exist at a common stage. Both sex and anatomical stage are represented by attributes. This formalization clarifies ontological structure and meaning and increases the capacity for formal reasoning about anatomy. The framework also supports generalizations such as vertebrate and invertebrate, thereby allowing the representation of anatomical structures that are common across a sub-phylum. PMID- 15797907 TI - GOR V server for protein secondary structure prediction. AB - SUMMARY: We have created the GOR V web server for protein secondary structure prediction. The GOR V algorithm combines information theory, Bayesian statistics and evolutionary information. In its fifth version, the GOR method reached (with the full jack-knife procedure) an accuracy of prediction Q3 of 73.5%. Although GOR V has been among the most successful methods, its online unavailability has been a deterrent to its popularity. Here, we remedy this situation by creating the GOR V server. PMID- 15797909 TI - Modularized learning of genetic interaction networks from biological annotations and mRNA expression data. AB - MOTIVATION: Inferring the genetic interaction mechanism using Bayesian networks has recently drawn increasing attention due to its well-established theoretical foundation and statistical robustness. However, the relative insufficiency of experiments with respect to the number of genes leads to many false positive inferences. RESULTS: We propose a novel method to infer genetic networks by alleviating the shortage of available mRNA expression data with prior knowledge. We call the proposed method 'modularized network learning' (MONET). Firstly, the proposed method divides a whole gene set to overlapped modules considering biological annotations and expression data together. Secondly, it infers a Bayesian network for each module, and integrates the learned subnetworks to a global network. An algorithm that measures a similarity between genes based on hierarchy, specificity and multiplicity of biological annotations is presented. The proposed method draws a global picture of inter-module relationships as well as a detailed look of intra-module interactions. We applied the proposed method to analyze Saccharomyces cerevisiae stress data, and found several hypotheses to suggest putative functions of unclassified genes. We also compared the proposed method with a whole-set-based approach and two expression-based clustering approaches. PMID- 15797908 TI - A practical false discovery rate approach to identifying patterns of differential expression in microarray data. AB - SUMMARY: Searching for differentially expressed genes is one of the most common applications for microarrays, yet statistically there are difficult hurdles to achieving adequate rigor and practicality. False discovery rate (FDR) approaches have become relatively standard; however, how to define and control the FDR has been hotly debated. Permutation estimation approaches such as SAM and PaGE can be effective; however, they leave much room for improvement. We pursue the permutation estimation method and describe a convenient definition for the FDR that can be estimated in a straightforward manner. We then discuss issues regarding the choice of statistic and data transformation. It is impossible to optimize the power of any statistic for thousands of genes simultaneously, and we look at the practical consequences of this. For example, the log transform can both help and hurt at the same time, depending on the gene. We examine issues surrounding the SAM 'fudge factor' parameter, and how to handle these issues by optimizing with respect to power. PMID- 15797910 TI - Genetic algorithm-based optimization of hydrophobicity tables. AB - SUMMARY: The genomic abundance and pharmacological importance of membrane proteins have fueled efforts to identify them based solely on sequence information. Previous methods based on the physicochemical principle of a sliding window of hydrophobicity (hydropathy analysis) have been replaced by approaches based on hidden Markov models or neural networks which prevail due to their probabilistic orientation. In the current study, an optimization of the hydrophobicity tables used in hydropathy analysis is performed using a genetic algorithm. As such, the approach can be viewed as a synthesis between the physicochemically and statistically based methods. The resulting hydrophobicity tables lead to significant improvement in the prediction accuracy of hydropathy analysis. Furthermore, since hydropathy analysis is less dependent on the basis set of membrane proteins is used to hone the statistically based methods, as well as being faster, it may be valuable in the analysis of new genomes. Finally, the values obtained for each of the amino acids in the new hydrophobicity tables are discussed. PMID- 15797912 TI - Correlation between gene expression profiles and protein-protein interactions within and across genomes. AB - MOTIVATION: Function annotation of an unclassified protein on the basis of its interaction partners is well documented in the literature. Reliable predictions of interactions from other data sources such as gene expression measurements would provide a useful route to function annotation. We investigate the global relationship of protein-protein interactions with gene expression. This relationship is studied in four evolutionarily diverse species, for which substantial information regarding their interactions and expression is available: human, mouse, yeast and Escherichia coli. RESULTS: In E.coli the expression of interacting pairs is highly correlated in comparison to random pairs, while in the other three species, the correlation of expression of interacting pairs is only slightly stronger than that of random pairs. To strengthen the correlation, we developed a protocol to integrate ortholog information into the interaction and expression datasets. In all four genomes, the likelihood of predicting protein interactions from highly correlated expression data is increased using our protocol. In yeast, for example, the likelihood of predicting a true interaction, when the correlation is > 0.9, increases from 1.4 to 9.4. The improvement demonstrates that protein interactions are reflected in gene expression and the correlation between the two is strengthened by evolution information. The results establish that co-expression of interacting protein pairs is more conserved than that of random ones. PMID- 15797911 TI - An algorithm for identification of bacterial selenocysteine insertion sequence elements and selenoprotein genes. AB - MOTIVATION: Incorporation of selenocysteine (Sec) into proteins in response to UGA codons requires a cis-acting RNA structure, Sec insertion sequence (SECIS) element. Whereas SECIS elements in Escherichia coli are well characterized, a bacterial SECIS consensus structure is lacking. RESULTS: We developed a bacterial SECIS consensus model, the key feature of which is a conserved guanosine in a small apical loop of the properly positioned structure. This consensus was used to build a computational tool, bSECISearch, for detection of bacterial SECIS elements and selenoprotein genes in sequence databases. The program identified 96.5% of known selenoprotein genes in completely sequenced bacterial genomes and predicted several new selenoprotein genes. Further analysis revealed that the size of bacterial selenoproteomes varied from 1 to 11 selenoproteins. Formate dehydrogenase was present in most selenoproteomes, often as the only selenoprotein family, whereas the occurrence of other selenoproteins was limited. The availability of the bacterial SECIS consensus and the tool for identification of these structures should help in correct annotation of selenoprotein genes and characterization of bacterial selenoproteomes. PMID- 15797913 TI - A robust neural networks approach for spatial and intensity-dependent normalization of cDNA microarray data. AB - MOTIVATION: Microarray experiments are affected by numerous sources of non biological variation that contribute systematic bias to the resulting data. In a dual-label (two-color) cDNA or long-oligonucleotide microarray, these systematic biases are often manifested as an imbalance of measured fluorescent intensities corresponding to Sample A versus those corresponding to Sample B. Systematic biases also affect between-slide comparisons. Making effective corrections for these systematic biases is a requisite for detecting the underlying biological variation between samples. Effective data normalization is therefore an essential step in the confident identification of biologically relevant differences in gene expression profiles. Several normalization methods for the correction of systemic bias have been described. While many of these methods have addressed intensity dependent bias, few have addressed both intensity-dependent and spatiality dependent bias. RESULTS: We present a neural network-based normalization method for correcting the intensity- and spatiality-dependent bias in cDNA microarray datasets. In this normalization method, the dependence of the log-intensity ratio (M) on the average log-intensity (A) as well as on the spatial coordinates (X,Y) of spots is approximated with a feed-forward neural network function. Resistance to outliers is provided by assigning weights to each spot based on how distant their M values is from the median over the spots whose A values are similar, as well as by using pseudospatial coordinates instead of spot row and column indices. A comparison of the robust neural network method with other published methods demonstrates its potential in reducing both intensity-dependent bias and spatial-dependent bias, which translates to more reliable identification of truly regulated genes. PMID- 15797915 TI - MADE4: an R package for multivariate analysis of gene expression data. AB - SUMMARY: MADE4, microarray ade4, is a software package that facilitates multivariate analysis of microarray gene-expression data. MADE4 accepts a wide variety of gene-expression data formats. MADE4 takes advantage of the extensive multivariate statistical and graphical functions in the R package ade4, extending these for application to microarray data. In addition, MADE4 provides new graphical and visualization tools that aid in interpretation of multivariate analysis of microarray data. PMID- 15797914 TI - Determining functional specificity from protein sequences. AB - MOTIVATION: Given a large family of homologous protein sequences, many methods can divide the family into smaller groups that correspond to the different functions carried out by proteins within the family. One important problem, however, has been the absence of a general method for selecting an appropriate level of granularity, or size of the groups. RESULTS: We propose a consistent way of choosing the granularity that is independent of the sequence similarity and sequence clustering method used. We study three large, well-investigated protein families: basic leucine zippers, nuclear receptors and proteins with three consecutive C2H2 zinc fingers. Our method is tested against known functional information, the experimentally determined binding specificities, using a simple scoring method. The significance of the groups is also measured by randomizing the data. Finally, we compare our algorithm against a popular method of grouping proteins, the TRIBE-MCL method. In the end, we determine that dividing the families at the proposed level of granularity creates very significant and useful groups of proteins that correspond to the different DNA-binding motifs. We expect that such groupings will be useful in studying not only DNA binding but also other protein interactions. PMID- 15797916 TI - Quasi-consensus-based comparison of profile hidden Markov models for protein sequences. AB - A simple approach for the sensitive detection of distant relationships among protein families and for sequence-structure alignment via comparison of hidden Markov models based on their quasi-consensus sequences is presented. Using a previously published benchmark dataset, the approach is demonstrated to give better homology detection and yield alignments with improved accuracy in comparison to an existing state-of-the-art dynamic programming profile-profile comparison method. This method also runs significantly faster and is therefore suitable for a server covering the rapidly increasing structure database. A server based on this method is available at http://liao.cis.udel.edu/website/servers/modmod PMID- 15797917 TI - Improved method for predicting beta-turn using support vector machine. AB - MOTIVATION: Numerous methods for predicting beta-turns in proteins have been developed based on various computational schemes. Here, we introduce a new method of beta-turn prediction that uses the support vector machine (SVM) algorithm together with predicted secondary structure information. Various parameters from the SVM have been adjusted to achieve optimal prediction performance. RESULTS: The SVM method achieved excellent performance as measured by the Matthews correlation coefficient (MCC = 0.45) using a 7-fold cross validation on a database of 426 non-homologous protein chains. To our best knowledge, this MCC value is the highest achieved so far for predicting beta-turn. The overall prediction accuracy Qtotal was 77.3%, which is the best among the existing prediction methods. Among its unique attractive features, the present SVM method avoids overtraining and compresses information and provides a predicted reliability index. PMID- 15797918 TI - Skin doses to patients undergoing coronary angiography in a Greek hospital. AB - In this study, the skin dose (SD) to patients undergoing coronary angiography (CA) were measured with thermoluminescence dosemeters (TLD) attached to various anatomical locations on the patient's skin during 93 CA procedures in a Greek hospital. The dose-area product (DAP) for every radiological projection was also measured for each procedure. The SD values were measured to be in the range 2.4 427.5 mGy, lower than the 2 Gy dose threshold for transient erythema. No general correlation was observed between the SD and the total DAP. PMID- 15797919 TI - Cytogenetic biodosimetry of an accidental exposure of a radiological worker using multiple assays. AB - A technician involved in the maintenance of X-ray equipment visited the occupational medicine service with complaints of skin lesions, apparently caused by an accidental exposure three months earlier. To estimate the dose received by the technician in the accident, biodosimetry was performed 6 and 18 months post exposure with the dicentric and micronucleus assays. Part of the latest blood sample was also used for retrospective dosimetry by fluorescence in situ hybridisation (FISH) analysis for translocations. The data obtained 6 and 18 months post-exposure indicate that both dicentrics and micronuclei disappear with a half-time of 1 y. After correction for delayed blood sampling, dose values of 0.75 Gy (95% confidence limits 0.56-1.05 Gy) from dicentrics and 0.96 Gy (95% confidence limits 0.79-1.18 Gy) from micronuclei were obtained. FISH analysis of translocations resulted in a dose estimate of 0.79 Gy (95% confidence limits 0.61 0.99 Gy). The satisfactory agreement between the three cytogenetic endpoints supports the use of the micronucleus assay for triage purposes in the case of large scale radiological accidents and provides further evidence for the valid use of FISH for translocations as a reliable retrospective biological dosimeter. PMID- 15797920 TI - Can default ICRP f1 values be applied to determine radiation dose from the intake of diet-incorporated thorium? AB - Data on the daily urinary excretion of thorium (Th) was obtained from 15 non exposed adult German subjects. A radiochemical neutron activation analysis method was developed and standardised especially for this purpose. The daily urinary excretion of 232Th was found to be in the range 1.9-14.9 microBq d(-1) with a mean (+/-SD) value of 6.5 (+/-4.3) microBq d(-1). Using this excretion value and reported data on dietary intake of Th for a similar German population, the gastrointestinal absorption factor (f1 value) proposed by the International Commission on Radiological Protection (ICRP) was tested. Although the daily excretion of 232Th observed in the present study was comparable to some of the currently reported values in certain other countries, it was higher than the excretion value calculated by applying the biokinetic model of Th proposed by ICRP for the dietary intake values. The study showed that the default ICRP values of the f1 factor for diet-incorporated Th may not be applicable. PMID- 15797921 TI - Occupational exposure to ionising radiation with thermoluminescence dosimetry system in Turkey, in 2003. AB - The individual annual dose information on classified workers who are occupationally exposed to extended radiation sources by using thermoluminescence dosimetry system, in Turkey, was assessed and analysed by the Ankara Nuclear Research and Training Centre (ANAEM) dosimetry service at the Turkish Atomic Energy Authority (TAEK) for the year 2003. A total of 3721 persons were monitored with TLD and the data presented in this report were obtained by using TLD technology in 2003. The annual mean effective doses received from external radiation in different fields of activities and the distribution of the annual effective dose by dose intervals are presented. The collective annual dose by field of activity is estimated and the contribution to the total annual collective dose is determined. PMID- 15797922 TI - A novel isoform of human Golgi complex-localized glycoprotein-1 (also known as E selectin ligand-1, MG-160 and cysteine-rich fibroblast growth factor receptor) targets differential subcellular localization. AB - The initial step in trafficking of leukocytes through the vascular endothelium is mediated by an adhesive interaction between molecules of the selectin family and their cognate receptors. Previously, a putative murine E-selectin ligand-1 (ESL 1) was identified and found to be identical to Golgi complex-localized glycoprotein-1 (GLG1), also known as MG-160, and to a previously identified basic fibroblast growth factor (bFGF)-binding protein known as cysteine-rich FGF receptor (CFR). We report here a novel variant of the human GLG1 gene product that we call GLG2, cloned from a human monocyte cDNA library. GLG2 encodes a polypeptide identical to GLG1 except with a unique 24-amino-acid extension at the C-terminus of its cytoplasmic domain. Transfection of chimeric constructs into human embryonic kidney epithelial 293 cells revealed that the cytoplasmic domains of GLG1 and GLG2 targeted the expression of each chimeric protein differentially, GLG1 to the cell surface and GLG2 to the Golgi. Genetic analysis suggests that GLG1 and GLG2 are the products of a single gene, the mRNA of which can be processed by alternative splicing to generate two different transcripts encoding either GLG1 or GLG2. Northern blot analysis showed that the relative amounts of the mRNAs for either isoform differ in a cell- and species-specific manner. These data suggest that alternative splicing of the GLG1 gene transcript might regulate the function of its product. PMID- 15797923 TI - Activation of either ERK1/2 or ERK5 MAP kinase pathways can lead to disruption of the actin cytoskeleton. AB - Oncogenic transformation often leads to the disruption of the actin cytoskeleton. Activation of the classical Ras-Raf-MEK1/2-ERK1/2 signalling cascade has been implicated in the effects of oncogenes such as Ras and Src on the cytoskeleton. Many of the studies of the effects of oncogenes on the cytoskeleton have made use of chemical inhibitors of MEK1/2 but it is now clear that these inhibitors also inactivate MEK5 in the MEK5-ERK5 MAP kinase pathway raising the possibility that this pathway may also be involved in oncogenic transformation. We therefore investigated whether activation of ERK5 can lead to disruption of the actin cytoskeleton. We show that activation of ERK5 can lead to loss of actin stress fibres, but by a distinct mechanism to ERK1/2. We demonstrate that ERK5 is activated by oncogenic Src as demonstrated by translocation of endogenous ERK5 from the cytoplasm to nucleus and activation of an ERK5-dependent transcriptional reporter and that ERK5 activation is required for Src-mediated transformation. We also show that in Src-transformed cells inhibition of ERK1/2 signalling is not sufficient for reappearance of the actin cytoskeleton and that ERK5 activation contributes to cytoskeletal disruption by Src. Our results suggest that multiple MAP kinase pathways downstream of oncogenes participate in cytoskeletal alterations. PMID- 15797924 TI - Mitomycin C-induced pairing of heterochromatin reflects initiation of DNA repair and chromatid exchange formation. AB - Chromatid interchanges induced by the DNA cross-linking agent mitomycin C (MMC) are over-represented in human chromosomes containing large heterochromatic regions. We found that nearly all exchange breakpoints of chromosome 9 are located within the paracentromeric heterochromatin and over 70% of exchanges involving chromosome 9 are between its homologues. We provide evidence that the required pairing of chromosome 9 heterochromatic regions occurs in G(0)/G(1) and S-phase cells as a result of an active cellular process initiated upon MMC treatment. By contrast, no pairing was observed for a euchromatic paracentromeric region of the equal-sized chromosome 8. The MMC-induced pairing of chromosome 9 heterochromatin is observed in a subset of cells; its percentage closely mimics the frequency of homologous interchanges found at metaphase. Moreover, the absence of pairing in cells derived from XPF patients correlates with an altered spectrum of MMC-induced exchanges. Together, the data suggest that the heterochromatin-specific pairing following MMC treatment reflects the initiation of DNA cross-link repair and the formation of exchanges. PMID- 15797925 TI - The nuclear rim protein Amo1 is required for proper microtubule cytoskeleton organisation in fission yeast. AB - Microtubules have a central role in cell division and cell polarity in eukaryotic cells. The fission yeast is a useful organism for studying microtubule regulation owing to the highly organised nature of its microtubular arrays. To better understand microtubule dynamics and organisation we carried out a screen that identified over 30 genes whose overexpression resulted in microtubule cytoskeleton abnormalities. Here we describe a novel nucleoporin-like protein, Amo1, identified in this screen. Amo1 localises to the nuclear rim in a punctate pattern that does not overlap with nuclear pore complex components. Amo1Delta cells are bent, and they have fewer microtubule bundles that curl around the cell ends. The microtubules in amo1Delta cells have longer dwelling times at the cell tips, and grow in an uncoordinated fashion. Lack of Amo1 also causes a polarity defect. Amo1 is not required for the microtubule loading of several factors affecting microtubule dynamics, and does not seem to be required for nuclear pore function. PMID- 15797926 TI - Assembly pathway of the anastral Drosophila oocyte meiosis I spindle. AB - Oocyte meiotic spindles of many species are anastral and lack centrosomes to nucleate microtubules. Assembly of anastral spindles occurs by a pathway that differs from that of most mitotic spindles. Here we analyze assembly of the Drosophila oocyte meiosis I spindle and the role of the Nonclaret disjunctional (Ncd) motor in spindle assembly using wild-type and mutant Ncd fused to GFP. Unexpectedly, we observe motor-associated asters at germinal vesicle breakdown that migrate towards the condensed chromosomes, where they nucleate microtubules at the chromosomes. Newly nucleated microtubules are randomly oriented, then become organized around the bivalent chromosomes. We show that the meiotic spindle forms by lateral associations of microtubule-coated chromosomes into a bipolar spindle. Lateral interactions between microtubule-associated bivalent chromosomes may be mediated by microtubule crosslinking by the Ncd motor, based on analysis of fixed oocytes. We report here that spindle assembly occurs in an ncd mutant defective for microtubule motility, but lateral interactions between microtubule-coated chromosomes are unstable, indicating that Ncd movement along microtubules is needed to stabilize interactions between chromosomes. A more severe ncd mutant that probably lacks ATPase activity prevents formation of lateral interactions between chromosomes and causes defective microtubule elongation. Anastral Drosophila oocyte meiosis I spindle assembly thus involves motor-associated asters to nucleate microtubules and Ncd motor activity to form and stabilize interactions between microtubule-associated chromosomes during the assembly process. This is the first complete account of assembly of an anastral spindle and the specific steps that require Ncd motor activity, revealing new and unexpected features of the process. PMID- 15797927 TI - Identification of the LEDGF/p75 HIV-1 integrase-interaction domain and NLS reveals NLS-independent chromatin tethering. AB - To investigate the basis for the LEDGF/p75 dependence of HIV-1 integrase (IN) nuclear localization and chromatin association, we used cell lines made stably deficient in endogenous LEDGF/p75 by RNAi to analyze determinants of its location in cells and its ability to interact with IN. Deletion of C-terminal LEDGF/p75 residues 340-417 preserved nuclear and chromatin localization but abolished the interaction with IN and the tethering of IN to chromatin. Transfer of this IN binding domain (IBD) was sufficient to confer HIV-1 IN interaction to GFP. HRP-2, the only other human protein with an identifiable IBD domain, was found to translocate IN to the nucleus of LEDGF/p75(-) cells. However, in contrast to LEDGF/p75, HRP-2 is not chromatin bound and does not tether IN to chromatin. A single classical nuclear localization signal (NLS) in the LEDGF/p75 N-terminal region ((146)RRGRKRKAEKQ(156)) was found by deletion mapping and was shown to be transferable to pyruvate kinase. Four central basic residues in the NLS are critical for its activity. Strikingly, however, stable expression studies with NLS(+/-) and IBD(+/-) mutants revealed that the NLS, although responsible for LEDGF/p75 nuclear import, is dispensable for stable, constitutive nuclear association of LEDGF/p75 and IN. Both wild-type LEDGF/p75 and NLS-mutant LEDGF/p75 remain entirely chromatin associated throughout the cell cycle, and each tethers IN to chromatin. Thus, these experiments reveal stable nuclear sequestration of a transcriptional regulator by chromatin during the nuclear cytosolic mixing of cell division, which additionally enables stable tethering of IN to chromatin. LEDGF/p75 is a multidomain adaptor protein that interacts with the nuclear import apparatus, lentiviral IN proteins and chromatin by means of an NLS, an IBD and additional chromatin-interacting domains. PMID- 15797928 TI - Endogenous Myc controls mammalian epidermal cell size, hyperproliferation, endoreplication and stem cell amplification. AB - The transcription factor Myc (c-Myc) plays an important role in cell growth and cell death, yet its physiological function remains unclear. Ectopic activation of Myc has been recently suggested to regulate cell mass, and Drosophila dmyc controls cellular growth and size independently of cell division. By contrast, it has been proposed that in mammals Myc controls cell division and cell number. To gain insights into this debate we have specifically knocked out Myc in epidermis. Myc epidermal knockout mice are viable and their keratinocytes continue to cycle, but they display severe skin defects. The skin is tight and fragile, tears off in areas of mechanical friction and displays impaired wound healing. Steady-state epidermis is thinner, with loss of the proliferative compartment and premature differentiation. Remarkably, keratinocyte cell size, growth and endoreplication are reduced, and stem cell amplification is compromised. The results provide new and direct evidence for a role for endogenous Myc in cellular growth that is required for hyperproliferative cycles and tissue homeostasis. PMID- 15797929 TI - Homologous and heterologous reconstitution of Golgi to chloroplast transport and protein import into the complex chloroplasts of Euglena. AB - Euglena complex chloroplasts evolved through secondary endosymbiosis between a phagotrophic trypanosome host and eukaryotic algal endosymbiont. Cytoplasmically synthesized chloroplast proteins are transported in vesicles as integral membrane proteins from the ER to the Golgi apparatus to the Euglena chloroplast. Euglena chloroplast preprotein pre-sequences contain a functional N-terminal ER-targeting signal peptide and a domain having characteristics of a higher plant chloroplast targeting transit peptide, which contains a hydrophobic stop-transfer membrane anchor sequence that anchors the precursor in the vesicle membrane. Pulse-chase subcellular fractionation studies showed that (35)S-labeled precursor to the light harvesting chlorophyll a/b binding protein accumulated in the Golgi apparatus of Euglena incubated at 15 degrees C and transport to the chloroplast resumed after transfer to 26 degrees C. Transport of the (35)S-labeled precursor to the chlorophyll a/b binding protein from Euglena Golgi membranes to Euglena chloroplasts and import into chloroplasts was reconstituted using Golgi membranes isolated from 15 degrees C cells returned to 26 degrees C. Transport was dependent upon extra- and intrachloroplast ATP and GTP hydrolysis. Golgi to chloroplast transport was not inhibited by N-ethylmaleimide indicating that fusion of Golgi vesicles to the chloroplast envelope does not require N ethylmaleimide-sensitive factor (NSF). This suggests that N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs) are not utilized in the targeting fusion reaction. The Euglena precursor to the chloroplast-localized small subunit of ribulose-1,5-bisphosphate carboxylase was not imported into isolated pea chloroplasts. A precursor with the N-terminal signal peptide deleted was imported, indicating that the Euglena pre-sequence has a transit peptide that functions in pea chloroplasts. A precursor to the small subunit of ribulose-1,5 bisphosphate carboxylase with the hydrophobic membrane anchor and the pre sequence region C-terminal to the hydrophobic membrane anchor deleted was imported localizing the functional transit peptide to the Euglena pre-sequence region between the signal peptidase cleavage site and the hydrophobic membrane anchor. The Euglena precursor to the small subunit of ribulose-1,5-bisphosphate carboxylase and the deletion constructs were not post-translationally imported into isolated Euglena chloroplasts indicating that vesicular transport is the obligate import mechanism. Taken together, these studies suggest that protein import into complex Euglena chloroplasts evolved by developing a novel vesicle fusion targeting system to link the host secretory system to the transit peptide dependent chloroplast protein import system of the endosymbiont. PMID- 15797930 TI - Ethnicity and variation in prognosis for patients newly hospitalised for heart failure: a matched historical cohort study. AB - OBJECTIVES: To compare mortality and factors predictive for outcome in age matched white and South Asian cohorts after first admission for heart failure. DESIGN: Matched historical cohort study. SETTING: One National Health Service trust comprising three acute care hospitals. PARTICIPANTS: 176 South Asian (mean age 68 (10) years, 45% women) and 352 age and sex matched white (70 (11) years, 42% women) patients hospitalised for the first time with heart failure. MAIN OUTCOME MEASURES: All cause survival, measures of disease severity, and the association of clinical variables with outcome. RESULTS: Compared with white patients, South Asian patients had similar rates of prior coronary heart disease but more often had prior hypertension (45% v 33%, p = 0.006) and diabetes (46% v 18%, p < 0.0001). Atrial fibrillation (15% v 31%, p = 0.0002) and prior diuretic use (39% v 48%, p = 0.041) were less common among South Asians. Left ventricular function was more often preserved (38% v 23%, p = 0.002) and less often severely impaired (18% v 28%, p = 0.025) among South Asians. During follow up (range 520 1880 days) 73 of 176 (41.2%) South Asian and 167 of 352 (47.4%) white patients died. South Asian ethnicity was associated with lower all cause mortality (odds ratio 0.71, 95% confidence interval 0.53 to 0.96, p = 0.02). Other predictors of outcome (admission age, lower systolic blood pressure, higher creatinine, higher plasma glucose, and lower haemoglobin) were similar in each cohort. CONCLUSIONS: At first hospitalisation, heart failure appears less advanced in South Asians, among whom diabetes and hypertension are more prevalent. Survival is better for South Asian than for white patients. Higher glucose and lower haemoglobin at admission provide useful prognostic information in heart failure. PMID- 15797932 TI - Aortic sclerosis: not an innocent murmur but a marker of increased cardiovascular risk. AB - The detection of an ejection systolic murmur in the aortic valve region often corresponds to a diagnosis of aortic sclerosis or minor disruption of the aortic valve with associated turbulence but minimal obstruction. Aortic sclerosis has two important clinical implications. Firstly, aortic sclerosis is an antecedent to clinically significant aortic valve obstruction and, secondly, it acts as a marker of increased risk of cardiovascular events. This article reviews the evidence that aortic sclerosis is a useful adjunctive tool in cardiovascular risk stratification and that its progression to haemodynamically significant aortic stenosis is a potential focus for individual monitoring and for interventional studies. PMID- 15797931 TI - Intravenous myocardial contrast echocardiography predicts regional and global left ventricular remodelling after acute myocardial infarction: comparison with low dose dobutamine stress echocardiography. AB - OBJECTIVE: To assess the role of intravenous myocardial contrast echocardiography (MCE) in predicting functional recovery and regional or global left ventricular (LV) remodelling after acute myocardial infarction (AMI) compared with low dose dobutamine stress echocardiography (LDSE). METHODS: 21 patients with anterior AMI and successful primary angioplasty underwent MCE and LDSE during the subacute stage (2-4 weeks after AMI). Myocardial perfusion and contractile reserve were assessed in each segment (12 segment model) with MCE and LDSE. The 118 dyssynergic segments in the subacute stage were classified as recovered, unchanged, or remodelled according to wall motion at six months' follow up. Percentage increase in LV end diastolic volume (%DeltaEDV) was also calculated. RESULTS: The presence of perfusion was less accurate than the presence of contractile reserve in predicting regional recovery (55% v 81%, p < 0.0001). However, the absence of perfusion was more accurate than the absence of contractile reserve in predicting regional remodelling (83% v 48%, p < 0.0001). The number of segments without perfusion was an independent predictor of %DeltaEDV, whereas the number of segments without contractile reserve was not. The area under the receiver operating characteristic curve showed that the number of segments without perfusion predicted substantial LV dilatation (%DeltaEDV > 20%) more accurately than did the number of segments without contractile reserve (0.88 v 0.72). CONCLUSION: In successfully revascularised patients with AMI, myocardial perfusion assessed by MCE is predictive of regional and global LV remodelling rather than of functional recovery, whereas contractile reserve assessed by LDSE is predictive of functional recovery rather than of LV remodelling. PMID- 15797933 TI - Effects of statins on endothelial function and lipid profile in HIV infected persons receiving protease inhibitor-containing anti-retroviral combination therapy: a randomised double blind crossover trial. PMID- 15797935 TI - Safety and cardiac chronotropic responsiveness to the early injection of atropine during dobutamine stress echocardiography in the elderly. AB - OBJECTIVE: To determine the safety and cardiac chronotropic responsiveness to early atropine dobutamine stress echocardiography (DSE) in the elderly. DESIGN: Retrospective study of 258 patients >or= 70 years who underwent early atropine DSE and 290 patients >or= 70 years who underwent conventional DSE. In the early atropine protocol, atropine was started at 20 microg/kg/min of dobutamine if heart rate was < 100 beats/min, up to 2 mg. The cardiac chronotropic responsiveness in the elderly was compared with a control group of patients < 70 years matched for sex, myocardial infarction, diabetes, and treatment with beta blockers and calcium channel blockers. RESULTS: The dose of dobutamine given to elderly patients was lower during early atropine than during conventional DSE (mean (SD) 29 (7) v 38 (4) microg/kg/min, p = 0.001). Early atropine DSE resulted in diminished incidence of ventricular extrasystoles, non-sustained ventricular tachycardia, bradycardia, and hypotension compared with conventional DSE. In comparison with patients < 70 years, elderly patients required lower doses of dobutamine and atropine and achieved a higher percentage of predicted maximum heart rate (92 (9)% v 88 (10)%, p = 0.0001). Except for more common hypotension (16% v 10%, p = 0.004), no other difference in adverse effects was observed between patients >or= 70 and < 70 years. CONCLUSIONS: Early atropine DSE is a safe strategy in the elderly resulting in lower incidence of minor adverse effects than with the conventional protocol. Elderly patients presented adequate cardiac chronotropic responsiveness to early injections of atropine, requiring lower doses of drugs to reach test end points. PMID- 15797934 TI - Effects of eplerenone on transcriptional factors and mRNA expression related to cardiac remodelling after myocardial infarction. AB - OBJECTIVE: To examine the effects of eplerenone, a selective aldosterone blocker, on cardiac function after myocardial infarction (MI) and myocardial remodelling related transcriptional factors and mRNA expression in non-infarcted myocardium. METHODS: MI was induced by ligation of the coronary artery in Wistar rats. Rats were randomly assigned to a vehicle treated group or an eplerenone treated group (100 mg/kg/day). RESULTS: At four weeks after MI, left ventricular (LV) end diastolic pressure, LV weight, and LV end diastolic dimension were increased in MI rats. Eplerenone significantly reduced the increase in LV end diastolic pressure, LV weight, and LV end diastolic dimension. In the MI rats the decreased ejection fraction indicated systolic dysfunction and the increased E wave to A wave ratio and E deceleration rate indicated diastolic dysfunction. Eplerenone significantly attenuated this systolic and diastolic dysfunction. Myocardial interstitial fibrosis, transcriptional activities of activator protein 1 and nuclear factor kappaB, and mRNA expression of monocyte chemoattractant protein 1, plasminogen activator inhibitor 1, atrial natriuretic peptide, brain natriuretic peptide, and collagen types I and III were significantly increased at four weeks after MI. Eplerenone significantly attenuated interstitial fibrosis and suppressed transcriptional activity and mRNA expression of these genes. CONCLUSIONS: When administered after MI, eplerenone prevents cardiac remodelling accompanied by systolic and diastolic dysfunction and inhibits abnormal myocardial transcriptional activities and gene expression. PMID- 15797936 TI - Validation of the EuroQol questionnaire in cardiac rehabilitation. AB - OBJECTIVE: To analyse the psychometric properties of the EuroQol questionnaire (EQ-5D) applied to patients with acute coronary syndromes (ACS). SETTING: Rehabilitation hospital. PATIENTS AND DESIGN: 106 consecutive patients with ACS (51% myocardial infarction, 42% coronary artery bypass grafting, 7% angina) completed the EQ-5D, the 36 item short form health survey (SF-36), and the MacNew questionnaire at admission, at discharge, and three months after inpatient cardiac rehabilitation. Acceptance, validity, reliability, and responsiveness of the EQ-5D were tested. RESULTS: The EQ-5D was highly accepted. The EQ-5D index showed substantial ceiling effects after rehabilitation. As expected the EQ-5D visual analogue scale (VAS) score (70.3 v 57.1) and EQ-5D index (77.8 v 64.5) were significantly better for patients with myocardial infarction than for patients who underwent surgery (both p < or = 0.001). Significant correlations were found between the EQ-5D VAS score, EQ-5D index, and domains of the SF-36 (r = 0.21 to r = 0.74). The correlation with the MacNew subscores and with the global score ranged between 0.55 and 0.78. With repeated measurement the EQ-5D showed reasonable reliability in stable patients with intraclass correlation ranging between 0.91 and 0.54. EQ-5D was responsive in patients who indicated improvement in health states between admission and discharge (effect size 0.74 0.82). CONCLUSION: The psychometric properties of the EQ-5D were satisfying. It is a reasonably valid, reliable, and responsive instrument for patients with ACS. It may be useful in clinical research and epidemiological studies to generate preference based valuations of health related quality life. PMID- 15797937 TI - The implantable cardioverter-defibrillator: postcode prescribing in the UK 1998 2002. AB - OBJECTIVE: To determine the rate of implantable cardioverter-defibrillator (ICD) implantation across the UK during the period 1998 to 2002. DESIGN: Observational self reporting with cross checking. SETTING: All ICD implanting centres coordinated by the National Pacemaker and ICD Database. PATIENTS: Every patient receiving an ICD in the UK from 1998 to 2002. MAIN OUTCOME MEASURES: Date of implantation and postcode of each ICD recipient during the study period. RESULTS: ICD implantation increased in the UK in the five year period studied but fell far short of the European average and national targets. Implantation rates varied greatly by region. CONCLUSIONS: The low rate of ICD implantation in the UK and the disparity between regions require further study to determine the barriers to this evidence based treatment. PMID- 15797939 TI - The stoichiometry and antenna size of the two photosystems in marine green algae, Bryopsis maxima and Ulva pertusa, in relation to the light environment of their natural habitat. AB - The stoichiometry and antenna sizes of the two photosystems in two marine green algae, Bryopsis maxima and Ulva pertusa, were investigated to examine whether the photosynthetic apparatus of the algae can be related to the light environment of their natural habitat. Bryopsis maxima and Ulva pertusa had chlorophyll (Chl) a/b ratios of 1.5 and 1.8, respectively, indicating large levels of Chl b, which absorbs blue-green light, relative to Chl a. The level of photosystem (PS) II was equivalent to that of PS I in Bryopsis maxima but lower than that of PS I in Ulva pertusa. Analysis of Q(A) photoreduction and P-700 photo-oxidation with green light revealed that >50% of PS II centres are non-functional in electron transport. Thus, the ratio of the functional PS II to PS I is only 0.46 in Bryopsis maxima and 0.35 in Ulva pertusa. Light-response curves of electron transport also provided evidence that PS I had a larger light-harvesting capacity than did the functional PS II. Thus, there was a large imbalance in the light absorption between the two photosystems, with PS I showing a larger total light harvesting capacity than PS II. Furthermore, as judged from the measurements of low temperature fluorescence spectra, the light energy absorbed by Chl b was efficiently transferred to PS I in both algae. Based on the above results, it is hypothesized that marine green algae require a higher ATP:NADPH ratio than do terrestrial plants to grow and survive under a coastal environment. PMID- 15797940 TI - Regulation of flooding tolerance of SAG12:ipt Arabidopsis plants by cytokinin. AB - A SAG12:ipt gene construct, which increases cytokinin biosynthesis in response to senescence, was introduced into Arabidopsis plants to delay senescence induced by flooding stress. Two forms of flooding stress, including total submergence and root waterlogging, were applied to SAG12:ipt (IPT) and wild-type (WT) plants for 1, 3, and 5 d. A separate experiment compared the recovery of WT and IPT plants subjected to flooding stress. Biomass accumulation, carbohydrate and chlorophyll contents, and cytokinin and abscisic acid were quantified to compare genotypic responses to flooding stress and post-flooding recovery. Real-time RT-PCR studies were performed to quantify ipt and SAG12 gene expression. IPT plants exposed to waterlogging accumulated greater quantities of cytokinins more rapidly than WT plants or those exposed to total submergence. Cytokinin accumulation was accompanied by phenotypic adaptations, including chlorophyll retention and increased biomass and carbohydrate content relative to WT plants. Abscisic acid accumulated rapidly in WT and IPT plants under waterlogging stress but remained low in all genotypes exposed to total submergence. IPT plants showed improved recovery after waterlogging stress was removed. Expression of ipt in submerged plants did not result in cytokinin accumulation until submergence stress was removed. At that point, IPT plants accumulated greater quantities of cytokinin and recovered to a greater extent than WT plants. This study established the relationship between flooding tolerance and cytokinin accumulation in IPT plants and suggested that translation of ipt transcripts and subsequent cytokinin accumulation were delayed under submergence stress. PMID- 15797938 TI - Superiority of sirolimus eluting stent compared with intracoronary beta radiation for treatment of in-stent restenosis: a matched comparison. AB - OBJECTIVE: To compare acute and follow up clinical and angiographic results after treatment of in-stent restenosis (ISR) by sirolimus eluting stents (SES) with results obtained after intracoronary radiation therapy (IRT). DESIGN: Matched pair analysis. METHODS: 62 consecutive ISR lesions (< 30 mm lesion length, reference diameter < 3.5 mm) in 62 patients were treated with SES. From a database of 174 lesions (n = 141 patients) treated for ISR by intracoronary beta radiation, 62 lesions (62 patients) were pair matched with the SES group for diabetes mellitus, lesion length, vessel size, and pattern of ISR. Six month angiographic and 12 month clinical follow up results were obtained. RESULTS: Baseline clinical and angiographic characteristics were similar between the groups (not significant). SES implantation resulted in significantly lower postprocedural in-lesion diameter stenosis than did IRT (mean (SD) 14.2 (9.5)% v 21.1 (10.6)%, p = 0.001), significantly higher minimum lumen diameter at follow up (1.91 (0.58) v 1.55 (0.72) mm, p = 0.005), and a higher net gain (1.16 (0.55) v 0.77 (0.70) mm, p = 0.002). Angiographic binary in-lesion restenosis rate at six months was 11% in the SES group and 29% in the IRT group (p = 0.046). In 16 ISR lesions SES were used after failed IRT and in 46 lesions for first time ISR. In-lesion late loss was higher after use of SES for failed IRT than after use of SES for first time ISR (0.61 (0.67) mm v 0.24 (0.41) mm, p = 0.018). In a multivariate analysis prior failed IRT was the only independent predictor for recurrent restenosis after SES for ISR (p = 0.052, odds ratio 5.8). Six patients (10%) in the SES group and 17 patients (27%) in the IRT group underwent target lesion revascularisation during the 12 months of follow up (p = 0.022). CONCLUSIONS: In this non-randomised matched cohort SES achieved acute and follow up results superior to IRT for treatment of ISR even if cases of failed IRT are included. Failed IRT is a predictor of impaired SES effectiveness. PMID- 15797946 TI - Abstracts of the Society for Experimental Biology Plant Frontier Meeting. PMID- 15797941 TI - Localization of aluminium in the maize root apex: can morin detect cell wall bound aluminium? AB - Morin is a fluorochrome which forms a fluorescent complex with aluminium (Al) and is thus used to localize Al in plant tissues. However, reports about the cellular distribution of Al-apoplastic versus symplastic-based on morin staining are often conflicting. The objective of this work was to investigate whether Al localization with morin staining can show the proper cellular distribution of Al. Fresh root cross-sections were made from root apices of maize (cv. Lixis) treated with 25 muM Al for 6 h and stained with morin. Fluorescence microscopic investigation showed Al-morin fluorescence in the cytosol, but not in the cell wall. This is in contrast to the growing evidence which shows that Al mainly accumulates in the cell wall, especially bound to the pectin matrix. Therefore, in vitro analyses were carried out to study whether morin can form a fluorescent complex with Al, which is bound to pectin, cell wall, and other Al-binding ligands such as phosphate, galacturonate, DNA, and ATP. Compared with the control treatment without Al-binding ligands, fluorescence intensity was reduced by about 10-fold in the presence of pectin and isolated cell walls, but fairly unaffected in the presence of phosphate and galacturonate. Al associated with DNA and ATP also formed a fluorescent complex with morin. This implies that, although Al is mainly accumulated in the cell wall, it cannot be detected with morin as it is tightly bound to cell-wall pectin. Thus, morin staining should not be used to study the distribution of Al between cell compartments. PMID- 15797947 TI - U.S. spending for mental health and substance abuse treatment, 1991-2001. AB - Spending for mental health and substance abuse (MHSA) treatment in the United States totaled dollar 104 billion in 2001, representing 7.6 percent of all health care spending. The nominal MHSA annual spending growth rate from 1991 to 2001 was 5.6 percent, almost one percentage point below the growth rate for all health care (6.5 percent). During this period, Medicaid has increased to be the largest payer of mental health care, with prescription drugs the fastest-growing spending component. Private insurance payment for substance abuse actually dropped in real dollars, increasing the public share of substance abuse spending. PMID- 15797948 TI - Plasma adiponectin in nonalcoholic fatty liver is related to hepatic insulin resistance and hepatic fat content, not to liver disease severity. AB - Plasma levels of adiponectin are decreased in patients with nonalcoholic fatty liver disease (NAFLD), but the relationship among plasma adiponectin, insulin sensitivity, and histological features is unclear. In 174 NAFLD patients and 42 controls, we examined plasma adiponectin concentrations in relation to 1) lipid profile, indices of insulin resistance, and features of the metabolic syndrome (n = 174); 2) hepatic insulin resistance (clamp technique with tracer infusion) (10 patients); and 3) histological features at liver biopsy (n = 116). When the data from all subjects were combined, plasma adiponectin levels were positively associated with increased age, female gender, and plasma high-density lipoprotein levels, and negatively associated with waist circumference, body mass index, triglycerides, indices of insulin resistance, and aminotransferase levels, and also predicted the presence of the metabolic syndrome. In step-wise regression, increased age, female gender, waist circumference, triglyceride levels, and homeostasis model assessment independently associated with adiponectin (adjusted R(2), 0.329). In NAFLD, adiponectin was only associated with increased age, female gender, and triglycerides (adjusted R(2), 0.245). When the measured histological parameters were included in the model, plasma adiponectin levels were also inversely proportional to the percentage of hepatic fat content (adjusted R(2), 0.221), whereas necroinflammation and fibrosis did not fit in the model. Adiponectin was negatively correlated with insulin-suppressed endogenous glucose production during the clamp (P = 0.011). The results demonstrate that decreased levels of circulating adiponectin in NAFLD are related to hepatic insulin sensitivity and to the amount of hepatic fat content. Hypoadiponectinemia in NAFLD is part of a metabolic disturbance characterized by ectopic fat accumulation in the central compartment. PMID- 15797949 TI - Detection of Turner syndrome using high-throughput quantitative genotyping. AB - CONTEXT: Turner syndrome (TS) is the most common genetic problem affecting women and occurs when an X chromosome is completely deleted, portions of an X chromosome are deleted, or chromosomal mosaicism occurs. Girls with TS may also have occult Y chromosome sequences. Whereas some girls with TS are identified in infancy or early childhood, many girls with TS are not detected until after 10 yr of age, resulting in delayed evaluation and treatment. OBJECTIVE: To prevent the delayed recognition and treatment of TS, a quantitative method of genotyping that can be performed as part of newborn screening is needed. DESIGN: To screen for sex chromosome abnormalities, we assembled a panel of informative single nucleotide polymorphism (SNP) markers that span the X chromosome from the dbSNP database. Pyrosequencing assays suitable for quantitative assessment of signal strength from single nucleotides were designed and used to genotype 46,XX; 46,XY; 45,X; and TS mosaics, examining zygosity and signal strength for individual alleles. Pyrosequencing assays were also designed for the detection of Y chromosome material. RESULTS: With just four informative SNP markers for the X chromosome, all TS girls with 45,X, partial X chromosome deletions, or mosaicism were identified with 100% sensitivity. In mosaic individuals, Y chromosomal material was detected with 100% sensitivity. CONCLUSION: These results suggest that inexpensive high-throughput screening is possible for TS and other sex chromosome disorders using quantitative genotyping approaches. PMID- 15797950 TI - Low-grade inflammation and estimates of insulin resistance during the menstrual cycle in lean and overweight women. AB - BACKGROUND: Inflammatory markers and insulin resistance are independent cardiovascular risk factors and are thought to be influenced by sex steroids. We investigated changes of inflammatory markers and estimates of insulin resistance during the menstrual cycle, their variances, and their relationship to each other, sex steroids, and regional body fat distribution. METHODS: Eight normal weight (body mass index, 21.6 +/- 1.9 kg/m(2)) and nine overweight (body mass index, 30 +/- 2.4 kg/m(2)) young women with normal ovarian function were assessed 15 times throughout the menstrual cycle. Regional fat distribution was assessed using dual x-ray absorptiometry. RESULTS: Concentrations of highly sensitive C reactive protein (hs-CRP) changed significantly during the menstrual cycle and were highest in the early follicular phase (P < 0.00001). SHBG concentrations were stable in the follicular phase but increased in the luteal phase (P < 0.00001). During the follicular phase, estimates of insulin resistance had a higher within-subject variance when determined by the homeostasis model assessment index (42%) than when estimated by SHBG concentrations (5%, P < 0.05). During the menstrual cycle, using repeated measurements, hs-CRP correlated inversely to estradiol (beta-coefficient, -0.23, P < 0.0001) and SHBG concentrations (beta-coefficient, -0.83, P = 0.004). Central accumulation of body fat correlated to the mean hs-CRP concentration (r = 0.63, P = 0.007) and the mean homeostasis model assessment index for insulin resistance (r = 0.75, P = 0.001). CONCLUSION: We demonstrate that serum concentrations of hs-CRP and SHBG significantly change during the menstrual cycle. We reveal a close link between sex steroids, subclinical inflammation, insulin resistance, and body fat distribution in regularly menstruating women. PMID- 15797951 TI - The Leu7Pro polymorphism of preproNPY is associated with decreased insulin secretion, delayed ghrelin suppression, and increased cardiovascular responsiveness to norepinephrine during oral glucose tolerance test. AB - CONTEXT: Neuropeptide Y (NPY) plays a role in angiogenesis, cardiovascular regulation, and hormone secretion. The leucine7 to proline7 (Leu7Pro) polymorphism of preproNPY is associated with vascular diseases and has an impact on hormone levels in healthy subjects. OBJECTIVE: The current study investigated the role of the Leu7Pro polymorphism in metabolic and cardiovascular autonomic regulation. DESIGN AND SUBJECTS: A 5-h oral glucose tolerance test was performed on 27 healthy volunteers representing two preproNPY genotypes (Leu7/Pro7 and Leu7/Leu7) matched for age, sex, body mass index and physical activity. MAIN OUTCOME MEASURES: Simultaneously we performed cardiovascular autonomic function tests and plasma measurements of sympathetic transmitters, glucose, insulin, and ghrelin. RESULTS: The subjects with Leu7/Pro7 genotype had decreased plasma NPY, norepinephrine (NE), and insulin concentrations and insulin to glucose ratios. The suppression of ghrelin concentrations after glucose ingestion was delayed in these subjects. They also had increased heart rate variability indices and baroreflex sensitivity. However, they displayed significant negative association of NE concentration with variability of low-frequency R-R-intervals and with baroreflex sensitivity. CONCLUSIONS: The Leu7Pro polymorphism of preproNPY is related to decreased level of basal sympathetic activity, decreased insulin secretion, and delayed ghrelin suppression during oral glucose tolerance test. The increased responsiveness of autonomic functions to NE associated with the polymorphism may be connected to increased cardiovascular vulnerability. PMID- 15797952 TI - Intrauterine exposure to diabetes is a determinant of hemoglobin A(1)c and systolic blood pressure in pima Indian children. AB - CONTEXT/OBJECTIVE: Given the increasing rates of both childhood obesity and type 2 diabetes (T2DM), we investigated whether maternal diabetes status during pregnancy is a determinant of risk factors associated with T2DM or cardiovascular disease in offspring during childhood. DESIGN/PARTICIPANTS: Forty-two Pima Indians, aged 7-11 yr, were identified retrospectively from maternal oral glucose tolerance tests as offspring of a diabetic pregnancy (22 ODM, eight males, 14 females) or offspring born before the mother developed diabetes (20 PRE, 12 males, eight females). SETTING/MAIN OUTCOME MEASURES: Weight, height, body mass index, percent body fat, blood pressure, and fasting concentrations of glucose, insulin, hemoglobin A1c (HbA1c), total cholesterol, triglycerides, and high density lipoprotein-cholesterol were measured while staying in an in-patient clinical research unit and compared in cross-sectional analyses. RESULTS: After adjustment for age and gender, ODM had significantly higher concentrations of HbA1c (ODM = 5.7 +/- 0.4, PRE = 5.0 +/- 0.4%, P = 0.002), higher systolic (SBP) blood pressure (ODM = 118 +/- 13, PRE = 107 +/- 10 mm Hg; P = 0.02), and lower concentrations of high-density lipoprotein (ODM = 41 +/- 9, PRE = 48 +/- 6 mg/dl, P = 0.03) than PRE. Maternal diabetes status during pregnancy persisted as a significant determinant of SBP (beta = 7.50, P = 0.03) and HbA1c (beta = 0.43, P = 0.002), independent of age, gender, and percent body fat. CONCLUSION: Intrauterine exposure to diabetes is a significant determinant of higher SBP and HbA1c during childhood, independent of adiposity and a genetic predisposition to T2DM. These data suggest that in utero exposure to diabetes confers an additional independent risk for the development of T2DM and/or cardiovascular disease later in life. PMID- 15797953 TI - Preproorexin and orexin receptors are expressed in cortisol-secreting adrenocortical adenomas, and orexins stimulate in vitro cortisol secretion and growth of tumor cells. AB - Orexins A and B are hypothalamic peptides that originate from the proteolytic cleavage of preproorexin and act through two subtypes of receptors, named OX1-R and OX2-R. OX1-R almost exclusively binds orexin-A, whereas OX2-R is nonselective for both orexins. We previously found that orexin-A, via the OX1-R, stimulates cortisol secretion from dispersed human adrenocortical cells. In this study, we demonstrate that six of eight cortisol-secreting adenomas expressed preproorexin mRNA, and seven of 10 adenomas contained measurable amounts of orexin-A but not orexin-B. Normal adrenal cortexes neither expressed preproorexin nor contained orexins. All adenomas expressed OX1-R and OX2-R mRNAs, and real-time PCR showed that the expression of both receptors was up-regulated in adenomas, compared with normal adrenal cortex. Orexin-A concentration-dependently raised basal cortisol secretion from freshly dispersed normal and adenomatous cells, minimal and maximal effective concentrations being 10(-10) and 10(-8) m, and the peptide efficacy (percent increase elicited by 10(-8) m orexin-A) was significantly higher in adenomas than in the normal adrenal cortex. Orexin-B was ineffective, thereby indicating that orexin secretagogue action is mediated by the OX1-R. In contrast, both orexins (10(-8) m) raised the proliferative activity of cultured normal and adenomatous cells, suggesting that this effect is mediated by OX2-R or both receptor subtypes. Collectively, our findings allow us to conclude that the orexin system is overexpressed in cortisol-secreting adenomas and suggest that orexin-A may act as an autocrine-paracrine regulator of the secretory activity and growth of some of these adrenal tumors. PMID- 15797954 TI - Prevalence of Vitamin D inadequacy among postmenopausal North American women receiving osteoporosis therapy. AB - PURPOSE: To evaluate serum 25-hydroxyvitamin D [25(OH)D] concentrations and factors related to vitamin D inadequacy in postmenopausal North American women receiving therapy to treat or prevent osteoporosis. METHODS: Serum 25(OH)D and PTH were obtained in 1536 community-dwelling women between November 2003 and March 2004. Multivariate logistic regression was used to assess risk factors for suboptimal (<30 ng/ml) 25(OH)D. RESULTS: Ninety-two percent of study subjects were Caucasian, with a mean age of 71 yr. Thirty-five percent resided at or above latitude 42 degrees north, and 24% resided less than 35 degrees north. Mean (sd) serum 25(OH)D was 30.4 (13.2) ng/ml: serum 25(OH)D was less than 20 ng/ml in 18%; less than 25 ng/ml in 36%; and less than 30 ng/ml in 52%. Prevalence of suboptimal 25(OH)D was significantly higher in subjects who took less than 400 vs. 400 IU/d or more vitamin D. There was a significant negative correlation between serum PTH concentrations and 25(OH)D. Risk factors related to vitamin D inadequacy included age, race, body mass index, medications known to affect vitamin D metabolism, vitamin D supplementation, exercise, education, and physician counseling regarding vitamin D. CONCLUSIONS: More than half of North American women receiving therapy to treat or prevent osteoporosis have vitamin D inadequacy, underscoring the need for improved physician and public education regarding optimization of vitamin D status in this population. PMID- 15797955 TI - The "obese insulin-sensitive" adolescent: importance of adiponectin and lipid partitioning. AB - There is a wide interindividual variation in peripheral insulin sensitivity at any given body mass index or percent body fat among obese adolescents with normal glucose tolerance. The goals of this study were to determine whether variability in insulin sensitivity is associated with differences in patterns of lipid partitioning or substrate use under fasting and hyperinsulinemic conditions. We compared 14 obese insulin-resistant adolescents with 14 obese insulin-sensitive controls, pair matched for age, gender, pubertal stage and body composition. Insulin sensitivity was assessed by the hyperinsulinemic-euglycemic clamp, intramyocellular lipid content by (1)H-nuclear magnetic resonance and visceral fat by magnetic resonance imaging. Obese insulin-sensitive subjects had lower intramyocellular (1.64 +/- 0.68 vs.2.26 +/- 0.62% of water peak, P = 0.017) and visceral lipid deposition (45 +/- 23 vs. 77 +/- 52 cm(2), P = 0.04) and a higher level of adiponectin, compared with their obese-resistant counterparts (8.8 +/- 3.6 vs. 6.5 +/- 1.8 mug/dl, P = 0.015). Glycerol fluxes were similar between the two obese groups yet occurred in the face of different concentrations of insulin. Intramyocellular lipid and visceral fat were negatively related to insulin sensitivity. Obese insulin-sensitive adolescents are characterized by lower lipid deposition in the intramyocellular and visceral compartments and greater levels of adiponectin, despite similar degree of adiposity. PMID- 15797957 TI - Tumor necrosis factor-alpha polymorphism, bone strength phenotypes, and the risk of fracture in older women. AB - TNFalpha is a proinflammatory cytokine that promotes osteoclastic bone resorption. We evaluated the association between a G-308A polymorphism (rs1800629) at the TNFA locus and osteoporosis phenotypes in 4306 older women participating in the Study of Osteoporotic Fractures. Femoral neck bone mineral density (BMD) and structural geometry were measured using dual-energy x-ray absorptiometry and hip structural analysis. Incident fractures were confirmed by physician adjudication of radiology reports. Despite similar femoral neck BMD, women with the A/A genotype had greater subperiosteal width (P = 0.01) and endocortical diameter (P = 0.03) than those with the G/G genotype. The net result of these structural differences was that there was a greater distribution of bone mass away from the neutral axis of the femoral neck in women with the A/A genotype, resulting in greater indices of bone bending strength (cross-sectional moment of inertia: P = 0.004; section modulus: P = 0.003). Among 376 incident hip fractures during 12.1 yr of follow-up, a 22% decrease in the risk of hip fracture was seen per copy of the A allele (relative risk 0.78; 95% confidence interval 0.63, 0.96), which was not influenced by adjustments for potential confounding factors, BMD, or bone strength indices. The G-308A polymorphism was not associated with a reduced risk of other fractures. These results suggest a potential role of genetic variation in TNFalpha in the etiology of osteoporosis. PMID- 15797956 TI - Age and uterine receptiveness: predicting the outcome of oocyte donation cycles. AB - CONTEXT: The impact of advancing age on uterine receptiveness has always been a concern of the medical establishment. Oocyte donation (OD) is the perfect model for ascertaining the extent of this relationship, but the literature is somewhat unreliable, mainly due to the limited samples on which the studies are based and insufficient control of important variables such as embryo quality. SETTING: The present work was developed in a private infertility clinic. PATIENTS OR OTHER PARTICIPANTS: We retrospectively evaluate the results of 3089 OD cycles that ended up in a d 3 embryo transfer. Severe male factor infertility was an exclusion criterion. MAIN OUTCOME MEASURES: The impact of patients' age on pregnancy, implantation, and miscarriage rates and obstetric outcome is analyzed, as is the relevance of endometrial thickness, serum estradiol levels, and duration of exogenous estrogen therapy to said rates. RESULTS: Pregnancy and implantation rates are significantly reduced and miscarriage rate is significantly increased from 45 yr of age onward. Concerning obstetric outcome, incidences of hypertension, proteinuria, premature rupture of membranes, second- and third-trimester hemorrhage, and preterm delivery are higher and mean birth weight is lower in this age group. With regard to endometrial preparation, estrogen therapy lasting more than 7 wk is associated with reduced PR and IR (P = 0.01 and P = 0.02, respectively). CONCLUSIONS: The results of OD cycles and obstetric outcome are significantly worse when recipients are 45 yr of age or older. Concerning endometrial preparation, results are significantly worse when estrogen therapy lasts more than 7 wk. PMID- 15797958 TI - Acute hyperglycemia does not affect the reactivity of coronary microcirculation in humans. AB - OBJECTIVE: There is some evidence that acute hyperglycemia (H) may cause vascular dysfunction in normal subjects. This study investigates whether acute, short-term H affects coronary vasodilatory function in healthy subjects. DESIGN: Diastolic peak flow velocity in the left anterior descending coronary artery was measured at rest and after dipyridamole (0.56 mg/kg over 4 min) using transthoracic color Doppler echocardiography in 13 healthy men. Coronary flow reserve (CFR) was defined as the ratio of dipyridamole-induced coronary peak diastolic to resting peak diastolic flow velocity. CFR was measured both in euglycemia (E) and after 3 h H ( approximately 14 mmol/liter) by a variable infusion of glucose and octreotide (0.4 mg/h) to prevent increase in insulin concentration. RESULTS: Fasting plasma glucose increased to 14.3 +/- 0.33 mmol/liter during the study and maintained variability within less than 10%. Plasma insulin remained nearly stable during H. Resting diastolic flow velocity was 18.5 +/- 0.6 cm/sec in E and increased to 20.0 +/- 0.7 cm/sec during H (P < 0.005). Dipyridamole infusion produced a marked increase in coronary flow velocity, which reached values of 50.8 +/- 2.9 cm/sec in E and 51.8 +/- 2.1 cm/sec in H (P = not significant). CFR was 2.78 +/- 0.16 in E and 2.59 +/- 0.12 in H (P = not significant). CONCLUSION: Our study indicates that short-term hyperglycemia does not affect the vasodilatory response of coronary microcirculation in healthy subjects. PMID- 15797959 TI - Prevention of bone loss after allogeneic stem cell transplantation by calcium, vitamin D, and sex hormone replacement with or without pamidronate. AB - CONTEXT: In controlled studies, bisphosphonates have been used to prevent bone loss after solid organ transplantations but not in conjunction with stem cell transplantation (SCT). OBJECTIVE: The objective of the study was to test whether additional iv pamidronate would prevent bone loss associated with SCT more effectively than the combination of calcium, vitamin D, and sex steroid replacement therapy alone. SETTING: The study was carried out at the Helsinki University Central Hospital. PATIENTS, DESIGN, INTERVENTION: Ninety-nine adult recipients of allogeneic SCT were randomized by age and gender into two groups. In one group, the patients received 1000 mg calcium carbonate and 800 IU vitamin D daily, and females received estrogen and males received testosterone replacement therapy. In another group, the patients received the same treatments plus six iv infusions of 60 mg pamidronate before and 1, 2, 3, 6, and 9 months after SCT. MAIN OUTCOME MEASURES: Bone mineral density (BMD) of the lumbar spine and the upper femur, measured by dual-energy x-ray absorptiometry, and bone turnover markers were followed for 12 months. RESULTS: In the pamidronate group, lumbar spine BMD remained stable but decreased in the other group by 2.9% at 12 months (P = 0.0084 between the groups over time). Total hip BMD reduced 5.1% in the pamidronate group and 7.8% in the other group by 12 months (P = 0.0015), and femoral neck BMD reduced 4.2 and 6.2%, respectively (P = 0.074). In the pamidronate group, serum type I procollagen amino-terminal propeptide (P = 0.032 between the groups over time) and urinary type I collagen amino-terminal telopeptide (P = 0.035) decreased 79 and 68% during the first 3 months, and remained lowered thereafter, but did not change in the other group. CONCLUSIONS: The recipients of allogeneic SCT receiving additional pamidronate sustain less bone loss than those treated with calcium, vitamin D, and sex steroid replacement alone. Despite all the efforts, however, bone loss is not totally abolished at the hip. PMID- 15797960 TI - Cutoff levels of 17-alpha-hydroxyprogesterone in neonatal screening for congenital adrenal hyperplasia should be based on gestational age rather than on birth weight. AB - OBJECTIVE: In newborn screening programs for congenital adrenal hyperplasia, 17 alpha-hydroxyprogesterone (17OHP) cutoff levels are based on birth weight (BW) or on gestational age (GA). We investigated which approach would result in the greatest specificity and sensitivity. STUDY DESIGN: For the determination of 17OHP, a neonatal 17OHP assay was used in filter paper blood of 9492 newborns. The relationships between 17OHP and BW and between 17OHP and GA were studied by regression analysis. Reference curves with a specificity of 99.95% were constructed with the method that summarizes the distribution by three smoothed curves representing the skewness (L curve), the median (M curve), and the coefficient of variation (S curve). Median cutoff levels for BW and for GA according to the 99.95% reference curves were calculated. RESULTS: Regression analysis showed that GA is a better predictor of 17OHP than BW (R(2) was 50.6 vs. 35.8%, respectively). At a specificity of 99.95%, the calculated median 17OHP cutoff level was lower for GA [12.6 microg/liter (38 nmol/liter)] than for BW [17.6 microg/liter (54 nmol/liter)], thus leading to a greater sensitivity. CONCLUSION: This study demonstrates that GA is a better predictor of 17OHP in newborns and will result in greater specificity than BW despite the fact that the determination of GA might be less reliable than BW. PMID- 15797961 TI - G protein receptors 7 and 8 are expressed in human adrenocortical cells, and their endogenous ligands neuropeptides B and w enhance cortisol secretion by activating adenylate cyclase- and phospholipase C-dependent signaling cascades. AB - Neuropeptides B and W (NPB and NPW) are regulatory peptides that act via two subtypes of G protein-coupled receptors, named GPR7 and GPR8. RT-PCR demonstrated the expression of these receptors in both zona glomerulosa and zona fasciculata reticularis (ZF/R) cells of the human adrenal cortex. NPB and NPW did not affect aldosterone secretion from dispersed zona glomerulosa cells but enhanced cortisol production from ZF/R cells, NPB being more effective than NPW. NPB evoked sizable cAMP and inositol triphosphate responses from ZF/R cells, which were abrogated by the adenylate cyclase inhibitor SQ-22536 and the phospholipase C inhibitor U 73122, respectively. Cortisol response to NPB was lowered by either SQ-22536 and the protein kinase (PK) A inhibitor H-89 or U-73122 and the PKC inhibitor calphostin-C and abolished by the simultaneous exposure to H-89 and calphostin-C. NPW elicited only a rise in cAMP production from dispersed ZF/R cells, and its cortisol response was suppressed by both SQ-22536 and H-89. PreproNPB and preproNPW mRNAs were detected in human adrenal cortexes. We conclude that: 1) NPB and NPW exert a secretagogue action on human ZF/R cells, probably acting in an autocrine-paracrine manner; and 2) the effect of NPB is mediated by both the adenylate cyclase/PKA and the phospholipase C/PKC cascades, whereas that of NPW involves only the activation of the former signaling pathway. PMID- 15797963 TI - The type 2 deiodinase A/G (Thr92Ala) polymorphism is associated with decreased enzyme velocity and increased insulin resistance in patients with type 2 diabetes mellitus. AB - The single-nucleotide polymorphism A/G in the type 2 deiodinase (D2) gene predicts a threonine (Thr) to alanine (Ala) substitution at codon 92 (D2 Thr92Ala) and is associated with insulin resistance in obese patients. Here, this association was investigated in 183 patients with type 2 diabetes mellitus, using homeostasis model assessment. The median fasting plasma insulin in Ala/Ala individuals was significantly higher than in patients with Ala/Thr or Thr/Thr genotypes (19.6 vs. 12.0 vs. 14.8 mIU/ml, respectively; P = 0.004). Assuming a recessive model, the homeostasis model assessment index was higher in the Ala/Ala group when compared with Ala/Thr-Thr/Thr group (8.50 vs. 4.85, P = 0.003). Although this polymorphism has not been associated with changes in D2 kinetics as measured in HEK-293 cells transiently expressing D2 Thr92Ala, we investigated whether such association could be detected in human tissue samples. Remarkably, in thyroid and skeletal muscle samples from subjects homozygous for the Ala allele, D2 velocity was significantly lower than in subjects with Ala/Thr-Thr/Thr genotypes (P = 0.05 and 0.04, respectively). In conclusion, the A/G polymorphism is associated with greater insulin resistance in type 2 diabetes mellitus patients and with lower D2 velocity in tissue samples. These findings suggest that the D2-generated T(3) in skeletal muscle plays a role in insulin resistance. PMID- 15797962 TI - Treatment with growth hormone and luteinizing hormone releasing hormone analog improves final adult height in children with congenital adrenal hyperplasia. AB - Final adult height is often compromised in children with congenital adrenal hyperplasia (CAH). This study examines the impact of GH and LHRH analog (LHRHa) on final adult height in patients with CAH due to 21-hydroxylase deficiency. Fourteen patients with CAH (eight males, six females) predicted to be more than 1.0 sd below their midparental target height received GH and LHRHa until final height. Each patient was matched at the start of GH therapy to a CAH patient treated only with glucocorticoids according to type of CAH, sex, and chronological age. Mean age, bone age, height, height prediction, and target height were the same in both groups at the beginning of GH therapy. Mean duration of GH treatment was 4.4 +/- 1.5 yr. Mean duration of LHRHa therapy was 4.2 +/- 2.0 yr. In the treatment group, final height sd score of -0.4 + 0.8 was significantly greater than both the initial prediction of -1.5 +/- 0.9 (P < 0.0001) and the final height sd score of the untreated group of -1.4 +/- 1.1 (P = 0.01). Our results indicate that the combination of GH and LHRHa improves final adult height in patients with CAH. PMID- 15797964 TI - Analysis of separate and combined effects of common variation in KCNJ11 and PPARG on risk of type 2 diabetes. AB - The separate and combined effects of the PPARG Pro(12)Ala polymorphism and the KCNJ11 Glu(23)Lys polymorphisms on risk of type 2 diabetes were investigated in relatively large-scale, case-control studies. Separate effects of the variants were examined among 1187/1461 type 2 diabetic patients and 4791/4986 middle-aged, glucose-tolerant subjects. The combined analysis involved 1164 type 2 diabetic patients and 4733 middle-aged, glucose-tolerant subjects. In the separate analyses, the K allele of the KCNJ11 Glu(23)Lys associated with type 2 diabetes (odds ratio, 1.19; P = 0.0002), whereas the PPARG Pro(12)Ala showed no significant association with type 2 diabetes. The combined analysis indicated that the two polymorphisms acted in an additive manner to increase the risk of type 2 diabetes, and we found no evidence for a synergistic interaction between them. Analysis of a model with equal additive effects of the two variants showed that the odds ratio for type 2 diabetes increased with 1.14/risk allele (P = 0.003). Together, the two polymorphisms conferred a population-attributable risk for type 2 diabetes of 28%. In conclusion, our results showed no evidence of a synergistic interaction between the KCNJ11 Glu(23)Lys and PPARG Pro(12)Ala polymorphisms, but indicated that they may act in an additive manner to increase the risk of type 2 diabetes. PMID- 15797966 TI - Effects of dehydroepiandrostenedione, superimposed on growth hormone substitution, on quality of life and insulin-like growth factor I in patients with secondary adrenal insufficiency: a randomized, placebo-controlled, cross over trial. AB - To assess whether dehydroepiandrostenedione (DHEA) substitution, superimposed on GH substitution, improves quality of life of patients with secondary adrenal failure, we studied the effects of DHEA (50 mg/d, 16 wk) vs. placebo (16 wk) in GH- and ACTH-deficient men (n = 15; age, 52 +/- 3 yr), and postmenopausal women (n = 16; age, 61 +/- 2 yr) in a double-blind, placebo-controlled, crossover study. All patients were receiving stable hormone replacement therapy, including a fixed dose of human recombinant GH during the study. The men received testosterone substitution. The female patients did not receive estrogen substitution. At baseline, multiple parameters of quality of life were impaired compared with age- and sex-matched controls, especially in female patients. These parameters were not improved by DHEA treatment. DHEA only slightly improved the depression score (women) and health perception (women and men), although these parameters were not abnormal at baseline. DHEA increased serum IGF-I concentrations in female patients (by approximately 18%; P < 0.001), but not in male patients. In neither group did DHEA affect IGF-binding protein-3 levels. We conclude that DHEA, superimposed on GH substitution, does not substantially improve quality of life in patients with secondary adrenal insufficiency regardless of gender. In addition, DHEA increases IGF-I levels only in estrogen depleted females, but not in testosterone-treated males, with secondary adrenal insufficiency. PMID- 15797965 TI - Changes in glucose tolerance over time in women with polycystic ovary syndrome: a controlled study. AB - We performed this study to access the changes in glucose tolerance over time in a group of women with polycystic ovary syndrome (PCOS) (n = 71) and control women (n = 23) with regular menstrual cycles and baseline normal glucose tolerance. Mean follow-up was between 2 and 3 yr for both groups (PCOS 2.5 +/- 1.7 yr; controls 2.9 +/- 2.1 yr). Based on World Health Organization glucose tolerance categories, there was no significant difference in the prevalence of glucose intolerance at follow-up in the PCOS group. In the PCOS group, 25 (37%) had impaired glucose tolerance (IGT) and seven (10%) had type 2 diabetes mellitus at baseline, compared with 30 (45%) and 10 (15%), respectively, at follow-up. There were also no differences within groups (PCOS or control) or between groups (PCOS vs. control) in the oral glucose tolerance test-derived measure of insulin sensitivity, but in the women with PCOS who converted to either IGT or type 2 diabetes mellitus, there was a significant decrease (P < 0.0001). At the follow up visit, the mean glycohemoglobin level was 6.1 +/- 0.9% in women with PCOS vs. 5.3 +/- 0.7% in the control women (P < 0.001). Women with PCOS and baseline IGT had a low conversion risk of 6% to type 2 diabetes over approximately 3 yr, or 2% per year. The effect of PCOS, given normal glucose tolerance (NGT) at baseline, is more pronounced with 16% conversion to IGT per year. Our study supports that women with PCOS (especially with NGT) should be periodically rescreened for diabetes due to worsening glucose intolerance over time, but this interval may be over several years and not annually. PMID- 15797967 TI - Virus-mediated oncolysis of thyroid cancer by a replication-selective adenovirus driven by a thyroglobulin promoter-enhancer region. AB - CONTEXT: Currently, there is no effective treatment for iodine-resistant thyroid cancers. OBJECTIVE: As a new approach to treatment, the efficacy of replication selective, human thyroglobulin (TG) enhancer and promoter-driven, adenovirus (AdhTGEP)-mediated oncolysis was investigated using two well-differentiated thyroid cancer cell lines, XTC (TG positive) and FTC-133 (TG negative), and other control tumor and nontumor cell lines (all TG negative). DESIGN: A cohort study design was used. SETTING: The study setting was laboratory bench-top experiments. SUBJECTS/PARTICIPANTS: In vitro TG-expressing and nonexpressing thyroid cell culture lines, nonthyroid tumor cell lines, as well as preclinical thyroid tumor bearing mice were studied. INTERVENTION: Adenoviral infection of cell lines was determined by immunohistochemistry, selective replication by one-step growth assays, and cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrozolium (MTS) assay. In vivo tumor growth inhibition was determined by a single intratumoral injection of 1 x 10(9) plaque-forming units AdhTGEP, AdLacZ (control virus), or PBS to 50- to 75 mm(3) tumors. XTC cells showed intense immunohistochemical staining, whereas FTC 133 and all other control cell lines showed minimal staining for viral infection with AdhTGEP. MAIN OUTCOME MEASURES: Cell survival and tumor growth inhibition after adenoviral infection were the main outcome measures. RESULTS: One-step growth assays showed at least a more than 60-fold titer of AdhTGEP in XTC than in FTC-133 cells. Cytotoxicity assays showed approximately 68% cell kill in XTC and minimal cell kill in FTC-133 and all other control cell lines at a multiplicity of infection of 250. There was significant in vivo growth inhibition of AdhTGEP treated XTC tumors (67 +/- 49 mm(3)) compared with AdLacZ-treated XTC (228 +/- 45 mm(3); P < 0.01), PBS-treated XTC (372 +/- 70 mm(3); P < 0.001), or AdhTGEP treated FTC-133 tumors (598 +/- 168 mm(3)). CONCLUSION: Replication-selective virus-mediated oncolysis is a potential therapy for recurrent, well differentiated, TG-secreting thyroid cancer that is unresponsive to standard treatment. PMID- 15797968 TI - Profiling of oxygen-modulated gene expression in early human placenta by systematic sequencing of suppressive subtractive hybridization products. AB - Villi from first-trimester human placenta were exposed to oxygen concentrations of either 2 or 20% during 3 h to construct two reciprocally subtracted libraries using the suppressive subtractive hybridization (SSH) methodology. After cloning, sequencing, and gene identification, the genes (1,071 clones corresponding to 822 different sequences) were classified according to 1) the subtracted library from which they originated and 2) within 58 groups of gene functions. We then developed a logarithm of the odds (LOD) test to identify a possible excess of genes in each group. We show that genes involved in angiogenesis are significantly overrepresented in the "hypoxic" condition (2% O2), whereas apoptotic genes are overrepresented in the "normoxic" condition (20% O2). Furthermore, we observed an excess of kinases relative to phosphatases and an excess of genes involved in proliferation over genes involved in cell growth in the hypoxic condition. To validate our results, we used quantitative RT-PCR to analyze the set of eight genes involved in angiogenesis on six independent placentas. Finally, we studied the distribution of gene clusters on human chromosomes to check whether their chromosomal distribution was random or not. We observed on human chromosome 11 a clear clustering of genes regulated similarly by O2 tension, and we also discovered indications that such clustering exists on chromosomes 6 and 12. PMID- 15797969 TI - Gene expression profiling of cerebellar development with high-throughput functional analysis. AB - We measured the expression levels of 450 genes during mouse postnatal cerebellar development by quantitative PCR using RNA purified from layers of the cerebellar cortex. Principal component analysis of the data matrix demonstrated that the first and second components corresponded to general levels of gene expression and gene expression patterns, respectively. We introduced 288 of the 450 genes into PC12 cells using a high-throughput transfection assay based on atelocollagen and determined the ability of each gene to promote neurite outgrowth or cell proliferation. Five genes induced neurite outgrowth, and seven genes enhanced proliferation. Evaluation of the functional data and gene expression patterns showed that none of these genes exhibited elevated expression at maturation, suggesting that genes characteristic of mature neurons are not likely to participate in neuronal development. These results demonstrate that functional data can facilitate interpretation of expression profiles and identification of new molecules that participate in biological processes. PMID- 15797970 TI - Alpha7-nicotinic acetylcholine receptor subunit is not required for parasympathetic control of the heart in the mouse. AB - Nicotinic acetylcholine receptors (nAChR) are assembled from a pool of nine alpha subunits and three beta-subunits into functional pentamers in peripheral autonomic neurons. The contribution of different subunits to native, physiologically important nAChR for synaptic transmission in autonomic ganglia is unclear. Here, we examined the importance of the alpha7-subunit for parasympathetic innervation of the heart. Normal (C57BL/6J), alpha7-deficient (Chrna7), and wild-type littermate mice were implanted with telemetry devices, and, under conscious, unsedated conditions, ECG recordings were obtained at baseline and after atropine, propranolol, and hexamethonium bromide administration. Spectral analysis of heart rate variability [power spectral analysis (PSA)] was performed for the evaluation of resting autonomic tone to the heart. At the completion of conscious studies, animals were anesthetized and underwent electrical stimulation of the vagus nerve (VS) while R-R intervals were recorded. Heart rate at baseline and after atropine, propranolol, or hexamethonium was similar in all three groups of animals. PSA curves were similar between normal, wild-type, and Chrna7 mice. VS showed no difference between control and Chrna7 mice throughout the range of stimulation (5-20 Hz). Mice deficient in the alpha7-nAChR subunit do not display differences in resting autonomic tone to the heart at baseline or under conditions of single and combined autonomic blockade. VS showed no difference in heart rate responses between normal and alpha7-deficient mice. These data support previous findings in vitro and highlight the important differences in function between nicotinic receptor subtypes because alpha3-deficient mice display major autonomic dysfunction. We conclude that the alpha7-subunit does not contribute critically to resting parasympathetic control of the heart. PMID- 15797971 TI - Cardiac transgenesis with the tetracycline transactivator changes myocardial function and gene expression. AB - The cardiac-specific tetracycline-regulated gene expression system (tet-system) is a powerful tool using double-transgenic mice. The cardiac alpha-myosin heavy chain promoter (alphaMHC) drives lifetime expression of a tetracycline-inhibited transcription activator (tTA). Crossing alphaMHC-tTA mice with mice containing a tTA-responsive promoter linked to a target gene yields double-transgenic mice having tetracycline-repressed expression of the target gene in the heart. Using the tet-system, some studies use nontransgenic mice for the control group, whereas others use single-transgenic alphaMHC-tTA mice. However, previous studies found that high-level expression of a modified activator protein caused cardiomyopathy. Therefore, we tested whether cardiac expression of tTA was associated with altered function of alphaMHC-tTA mice compared with wild-type (WT) littermates. We monitored in vivo and in vitro function and gene expression profiles for myocardium from WT and alphaMHC-tTA mice. Compared with WT littermates, alphaMHC-tTA mice had a greater heart-to-body weight ratio (approximately 10%), ventricular dilation, and decreased ejection fraction, suggesting mild cardiomyopathy. In vitro, submaximal contractions were greater compared with WT and were associated with greater myofilament Ca2+ sensitivity. Gene expression profiling revealed that the expression of 153 genes was significantly changed by >20% when comparing alphaMHC-tTA with WT myocardium. These findings demonstrate that introduction of the alphaMHC-tTA construct causes significant effects on myocardial gene expression and major functional abnormalities in vivo and in vitro. For studies using the tet-system, these results suggest caution in the use of controls, since alphaMHC-tTA myocardium differs appreciably from WT. Furthermore, the results raise the possibility that the phenotype conferred by a target gene may be influenced by the modified genetic background of alphaMHC-tTA myocardium. PMID- 15797974 TI - Water balance in rheumatoid arthritis. PMID- 15797972 TI - Increased cutaneous T-cell-attracting chemokine levels in sera from patients with systemic sclerosis. AB - OBJECTIVE: We sought to assess serum cutaneous T-cell-attracting chemokine (CTACK) levels and CTACK expression levels in skin from patients with systemic sclerosis (SSc), and determine whether serum CTACK levels correlate with clinical features in SSc patients. METHODS: Serum samples were obtained from 73 SSc patients, 32 patients with systemic lupus erythematosus and 26 patients with dermatomyositis. Serum CTACK levels were determined by enzyme-linked immunoabsorbent assay. CTACK mRNA expression in sclerotic skin was assessed by real-time reverse transcription-polymerase chain reaction. RESULTS: Serum CTACK levels were significantly increased in patients with diffuse cutaneous SSc (dSSc; n=32) and those with limited cutaneous SSc (lSSc; n=41) compared with normal controls (n=31; P<0.05 and P<0.0005, respectively). The presence of calcinosis and muscle involvement was more frequently detected in SSc patients with elevated CTACK levels (P<0.05 and P<0.05, respectively). Elevated C-reactive protein levels were also observed more frequently in SSc patients with increased CTACK levels (P<0.05). CTACK mRNA expression levels in the sclerotic skin of SSc patients were augmented. In a longitudinal study, serum CTACK levels were generally decreased during the follow-up. CONCLUSIONS: The increased serum CTACK levels and enhanced skin CTACK expression in SSc patients suggest that CTACK is related to the inflammation associated with SSc. PMID- 15797976 TI - Methotrexate in pregnancy. PMID- 15797975 TI - Systemic lupus erythematosus in a multiethnic US Cohort (LUMINA). XXX: association between C-reactive protein (CRP) gene polymorphisms and vascular events. AB - OBJECTIVES: To determine if a polymorphic GTn repeat in the intron of the C reactive protein (CRP) gene associates with occurrence of vascular arterial events in systemic lupus erythematosus (SLE). METHODS: We performed a nested case control study on the LUMINA cohort of 546 Hispanic, African-American and Caucasian SLE patients. Twenty-five patients who developed vascular arterial events (i.e. myocardial infarction, angina, coronary artery bypass graft surgery, stroke, claudication, gangrene or significant tissue loss and/or arterial peripheral thrombosis) after enrolment were selected as cases and 32 ethnically matched patients with no previous vascular arterial events served as controls. Their CRP gene GTn polymorphism and plasma CRP was determined. RESULTS: Patients with vascular events had more severe SLE and were more likely to have plasma CRP in the highest quintile of measured values. The overall distribution of GTn alleles for patients with vascular events had a greater number of the GT20 variant compared with controls [26.0% of alleles (13/50) vs 15.6% (10/64)]. This greater number of GT20 in patients with vascular events was observed for African Americans [29.2% (7/24) vs 21.0% (8/38)] and Hispanics [33.0% (4/12) vs 0% (0/16)] but not for Caucasians [14.3% (2/14) vs 20.0% (2/10)]. For African Americans and Hispanics combined (45 patients), the frequency of GT20 in those with vascular events (30.6%, 11/36) was significantly higher than in those without them (14.8%, 8/54) (P<0.05, one-tailed test for difference in proportions). When patients were categorized according to the number of GT20 alleles they carried (thus GT20/GT20, GT20/GTx or GTx/GTx, where x is any allele other than GT20), for both African-Americans and Hispanics the likelihood of vascular arterial events increased in proportion with the GT20 dose, and all GT20 homozygous patients developed vascular arterial events. CONCLUSIONS: The CRP GT20 variant is more likely to occur in African-American and Hispanic SLE patients than in Caucasian ones, and SLE patients carrying the GT20 allele are more likely to develop vascular arterial events. PMID- 15797978 TI - Translational research in autoimmunity: aims of therapy in vasculitis. PMID- 15797979 TI - Hand surgery in rheumatoid arthritis: state of the art and suggestions for research. AB - OBJECTIVE: The role of surgery in the clinical management of patients with rheumatoid arthritis (RA)-associated hand dysfunction is still a subject of controversy. The efficacy of surgery in RA-associated hand dysfunction is assessed through an exhaustive review of published studies. METHODS: A high sensitivity search strategy was used to identify in MedLine and CENTRAL original studies related to hand and wrist surgery in RA patients. We selected articles including at least two adult RA patients which evaluated clinical outcomes through an observational or experimental design. Eligible studies were evaluated by standardized criteria. Two investigators independently used a pre-defined form to extract data about patient population, intervention, follow-up and clinical outcomes. Disagreements were discussed and resolved. RESULTS: One hundred and ninety-six papers met inclusion criteria. Only five were randomized trials, while most studies followed an observational design, often of poor quality. As such, we could not pool data for statistical analysis; however, we were still able to provide a best evidence synthesis. A positive trend suggesting the efficacy of total carpal arthrodesis and metacarpophalangeal arthroplasty in reducing pain and improving function seemed to emerge from the published studies. CONCLUSIONS: Despite recent advances in medical treatment, surgery still plays a role in the clinical management of RA-associated hand dysfunction. However, the majority of the available studies showed methodological flaws that prevented a clear definition of both surgical indications and criteria for choosing any specific procedure. Suggestions for further investigations are also provided. PMID- 15797980 TI - Development and preliminary validation of a systemic lupus erythematosus-specific quality-of-life instrument (SLEQOL). AB - OBJECTIVES: Systemic lupus erythematosus (SLE), a chronic illness with an unpredictable and variable course, profoundly affects the quality of life (QOL). General health questionnaires are used to assess QOL in SLE, but a disease specific instrument could offer enhanced responsiveness and content validity. We detail the steps we took to develop and validate a new SLE-specific QOL instrument, SLEQOL. METHODS: Rheumatology professionals nominated items that they felt were important determinants of QOL of SLE patients. One hundred SLE patients were asked to assess the importance and frequency of occurrence of these items and to suggest those that had not been listed. Item reduction was performed using Rasch model and factor analyses to create a new questionnaire in English. This final questionnaire was administered to a cohort of 275 patients to study its psychometric properties. RESULTS: Fifty-one items covering a wide range of QOL concerns were identified. The patients' responses led to the elimination of 11. The new questionnaire of 40 items was found to have Cronbach's alpha of 0.95 and to consist of eight domains covering physical, mental and social QOL issues. It has good test-retest reliability, poor to fair cross-sectional correlation with the SF-36, with poor correlation with lupus activity or damage indices. The SLEQOL was more responsive to change than the SF-36. CONCLUSIONS: We have developed a new 40-item SLEQOL in English and showed that it is valid for use in SLE patients in Singapore. It offers better content validity and responsiveness to change than the SF-36. PMID- 15797982 TI - Efficacy of topical anesthetics to reduce pain in premature infants during eye examinations for retinopathy of prematurity. AB - BACKGROUND: Eye examinations for retinopathy of prematurity (ROP) are stressful and probably painful, but many ophthalmologists do not apply topical anesthetics because their efficacy in reducing pain has not been established. OBJECTIVE: To evaluate the potential benefits of topical anesthetic eye drops in reducing pain during neonatal eye examination for ROP. METHODS: Neonates born at < or =30 weeks' gestation and expected to have at least 2 examinations for ROP were included. Patients were randomly assigned to receive either proparacaine HCl ophthalmic solution 0.5% or NaCl 0.9% (saline) eye drops prior to an eye examination. In a subsequent examination, each patient received the alternate treatment. Eye drops were prepared in the pharmacy in identical tuberculin syringes, and physicians, nurses, and pharmacists were blinded to the treatment given. Pain was measured using a scoring system with both physical and physiologic measures of pain (Premature Infant Pain Profile [PIPP], possible range 1-21), which has been validated in preterm infants. PIPP scoring was performed simultaneously by 2 nurses: 1 and 5 minutes before and after the eye examination and during initial placement of the eye speculum. The same ophthalmologist performed all examinations. RESULTS: Twenty-two patients were studied, with 11 infants receiving proparacaine and 11 receiving saline as the first treatment. Crossover was performed with a median of 17.5 days between treatments. Patients experienced significantly less pain at speculum insertion with proparacaine than with saline (paired difference -2.5 +/- 3.4; p = 0.001). CONCLUSIONS: Topical anesthetic pretreatment reduces the pain response to eye examination for ROP and should become routine practice. Because this is not effective in all infants, additional measures to reduce pain should be taken. PMID- 15797981 TI - Blood pressure in Native Americans switched from celecoxib to rofecoxib. AB - BACKGROUND: Nonsteroidal antiinflammatory drugs have been associated with exacerbation of hypertension. Differing effects on blood pressure (BP) have been reported in studies comparing celecoxib and rofecoxib. Concern regarding the cardiovascular safety of the cyclooxygenase-2 (COX-2) inhibitor class has intensified since the removal of rofecoxib from the market. OBJECTIVE: To evaluate the effect of a formulary change from celecoxib to rofecoxib on the BP of Native American patients at an Indian Health Service medical center. METHODS: Medical records of patients switched from celecoxib to rofecoxib were retrospectively reviewed. BP during the respective treatments was compared as follows: measurements recorded while taking celecoxib within 6 months before the index date and while taking rofecoxib from 1 week after the index date through 6 months of treatment were averaged. Differences in systolic and diastolic BP before and after the therapy change were evaluated using a paired Student's t test. Subgroup analysis was performed for patients with preexisting hypertension. RESULTS: During rofecoxib therapy, the mean systolic BP was 2.9 mm Hg higher (p = 0.015) and the mean diastolic BP was 1.5 mm Hg higher (p = 0.042) than during celecoxib therapy. Among hypertensive patients, the respective mean systolic and diastolic BPs were 4.8 mm Hg (p = 0.009) and 2.0 mm Hg (p = 0.063) higher while taking rofecoxib. CONCLUSIONS: Switching patients from celecoxib to rofecoxib resulted in an increase in BP, with a larger difference observed in patients with hypertension. Future studies assessing the cardiovascular safety of currently marketed and investigational COX-2 inhibitors should evaluate the possible contribution of BP effects of these agents to overall risk. PMID- 15797983 TI - Update on personal digital assistant applications for the healthcare provider. AB - OBJECTIVE: To review the common general drug information applications and specialty drug information applications available for personal digital assistants (PDAs). DATA SOURCES: The literature was accessed through MEDLINE (2003-June 2004). Other information was obtained through secondary sources, such as Web sites describing common PDA applications as well as actual product trials. The key search terms used were handheld, PDA, personal digital assistants, drug information, pharmacokinetics, medical information, and medical applications. STUDY SELECTION AND DATA EXTRACTION: Articles or studies that provided a review of drug information references for the PDA since 2002 were included. Data pertaining to cost and application size were obtained from product or vendor Web sites. DATA SYNTHESIS: There are numerous medical applications available for the PDA including general drug information references, specialty drug information references (eg, pediatrics, cardiology, infectious diseases, oncology, psychology, herbals), diagnostic applications, medical calculators, nursing references, and patient tracking databases. Due to the huge array of programs, as well as factors such as cost and memory requirements, the healthcare provider must be selective in the medical applications that are placed on the PDA. CONCLUSIONS: There are many excellent PDA drug information applications that provide fast and accurate drug information and other features that assist the healthcare provider. PMID- 15797984 TI - Comment: current options in the management of olanzapine-associated weight gain. PMID- 15797985 TI - Effect of murine strain on metabolic pathways of glucose production after brief or prolonged fasting. AB - Background strain is known to influence the way a genetic manipulation affects mouse phenotypes. Despite data that demonstrate variations in the primary phenotype of basic inbred strains of mice, there is limited data available about specific metabolic fluxes in vivo that may be responsible for the differences in strain phenotypes. In this study, a simple stable isotope tracer/NMR spectroscopic protocol has been used to compare metabolic fluxes in ICR, FVB/N (FVB), C57BL/6J (B6), and 129S1/SvImJ (129) mouse strains. After a short-term fast in these mice, there were no detectable differences in the pathway fluxes that contribute to glucose synthesis. However, after a 24-h fast, B6 mice retain some residual glycogenolysis compared with other strains. FVB mice also had a 30% higher in vivo phosphoenolpyruvate carboxykinase flux and total glucose production from the level of the TCA cycle compared with B6 and 129 strains, while total body glucose production in the 129 strain was approximately 30% lower than in either FVB or B6 mice. These data indicate that there are inherent differences in several pathways involving glucose metabolism of inbred strains of mice that may contribute to a phenotype after genetic manipulation in these animals. The techniques used here are amenable to use as a secondary or tertiary tool for studying mouse models with disruptions of intermediary metabolism. PMID- 15797986 TI - Endogenous effectors of human liver glycogen phosphorylase modulate effects of indole-site inhibitors. AB - Phosphorylase is regulated by a number of small-molecular-weight effectors that bind to three sites on the enzyme. Recently, a fourth site referred to as the indole-inhibitor site has been identified. Synthetic compounds bind to the site and inhibit activity. However, the effects of these compounds in the presence of other endogenous effectors are unknown. We have determined the effects of four indole derivative glycogen phosphorylase inhibitors (GPI) on recombinant human liver glycogen phosphorylase a activity. The GPIs tested were all potent inhibitors. However, the endogenous inhibitors (glucose, ADP, ATP, fructose 1 phosphate, glucose 6-phosphate, UDP-glucose) and the activator (AMP) markedly reduced the inhibitory effect of GPIs. Consistent with these in vitro findings, the IC50 for the inhibition of glycogenolysis in cells and the liver drug concentration associated with glucose-lowering activity in diabetic ob/ob mice in vivo were also significantly higher than those determined in in vitro enzyme assays. The inhibitory effect of indole-site effectors is modulated by endogenous small-molecular-weight effectors of phosphorylase a activity. However, at higher concentrations (10-30 microM), the GPI effect was dominant and resulted in inhibition of phosphorylase a activity irrespective of the presence or absence of the other modulators of the enzyme. PMID- 15797987 TI - Alterations in carbohydrate metabolism in response to short-term dietary carbohydrate restriction. AB - Dietary carbohydrate restriction (CR) presents a challenge to glucose homeostasis. Despite the popularity of CR diets, little is known regarding the metabolic effects of CR. The purpose of this study was to examine changes in whole body carbohydrate oxidation, glucose availability, endogenous glucose production, and peripheral glucose uptake after dietary CR, without the confounding influence of a negative energy balance. Postabsorptive rates of glucose appearance in plasma (R(a); i.e., endogenous glucose production) and disappearance from plasma (R(d); i.e., glucose uptake) were measured using isotope dilution methods after a conventional diet [60% carbohydrate (CHO), 30% fat, and 10% protein; kcals = 1.3 x resting energy expenditure (REE)] and after 2 days and 7 days of CR (5% CHO, 60% fat, and 35% protein; kcals = 1.3 x REE) in eight subjects (means +/- SE; 29 +/- 4 yr; BMI 24 +/- 1 kg/m(2)) during a 9-day hospital visit. Postabsorptive plasma glucose concentration was reduced (P = 0.01) after 2 days but returned to prediet levels the next day and remained at euglycemic levels throughout the diet (5.1 +/- 0.2, 4.3 +/- 0.3, and 4.8 +/- 0.4 mmol/l for prediet, 2 days and 7 days, respectively). Glucose R(a) and glucose R(d) were reduced to below prediet levels (9.8 +/- 0.6 micromol x kg(-1) x min( 1)) after 2 days of CR (7.9 +/- 0.3 micromol x kg(-1) x min(-1)) and remained suppressed after 7 days (8.3 +/- 0.4 micromol x kg(-1) x min(-1); both P < 0.001). A greater suppression in carbohydrate oxidation, compared with the reduction in glucose R(d), led to an increased (all P T, MTHFR 1298A>C), methionine synthase (MTR 2756A>G), methionine synthase reductase (MTRR 66A>G), methylenetetrahydrofolate dehydrogenase (MTHFD1 1958G>A), serine hydroxymethyl transferase (SHMT1 1420C>T), thymidylate synthase (TS 2R3R), and the reduced folate carrier (RFC 80G>A) were detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) or real-time PCR. Patients with the MTHFR 1298AC variant or the MTRR 66 G-allele showed decreased in vitro MTX sensitivity measured under both test conditions. SHMT1 1420TT homozygotes only showed decreased MTX sensitivity in the TSI(50, cont). In conclusion, polymorphisms in the folate-related genes MTHFR, MTRR, and SHMT1 are related to MTX resistance in pediatric patients with ALL. PMID- 15797995 TI - Transforming growth factor-{beta}1 modulates responses of CD34+ cord blood cells to stromal cell-derived factor-1/CXCL12. AB - Disruption of stromal cell-derived factor-1 (SDF-1/CXCL12 [CXC chemokine ligand 12]) interaction leads to mobilization of stem/progenitor cells from bone marrow to circulation. However, prolonged exposure of CD34+ cells to SDF-1 desensitizes them to SDF-1. So how do cells remain responsive to SDF-1 in vivo when they are continuously exposed to SDF-1? We hypothesized that one or more mechanisms mediated by cytokines exist that could modulate SDF-1 responsiveness of CD34+ cells and the desensitization process. We considered transforming growth factor beta1 (TGF-beta1) a possible candidate, since TGF-beta1 has effects on CD34+ cells and is produced by stromal cells, which provide niches for maintenance and proliferation of stem/progenitor cells. TGF-beta1 significantly restored SDF-1 induced chemotaxis and sustained adhesion responses in cord blood CD34+ cells preexposed to SDF-1. Effects of TGF-beta1 were dependent on the dose and duration of TGF-beta1 pretreatment. Phosphorylation of extracellular signal-regulated kinase 1 (Erk1)/Erk2 was implicated in TGF-beta1 modulation of migratory and adhesion responses to SDF-1. Our results indicate that low levels of TGF-beta1 can modulate SDF-1 responsiveness of CD34+ cells and thus may facilitate SDF-1 mediated retention and nurturing of stem/progenitor cells in bone marrow. PMID- 15797996 TI - Critical role of host gammadelta T cells in experimental acute graft-versus-host disease. AB - Gammadelta T cells localize to target tissues of graft-versus-host disease (GVHD) and therefore we investigated the role of host gammadelta T cells in the pathogenesis of acute GVHD in several well-characterized allogeneic bone marrow transplantation (BMT) models. Depletion of host gammadelta T cells in wild-type (wt) B6 recipients by administration of anti-T-cell receptor (TCR) gammadelta monoclonal antibody reduced GVHD, and gammadelta T-cell-deficient (gammadelta-/-) BM transplant recipients experienced markedly improved survival compared with normal controls (63% vs 10%, P < .001). gammadelta T cells were responsible for this difference because reconstitution of gammadelta-/- recipients with gammadelta T cells restored GVHD mortality. gammadelta-/- recipients showed decreased serum levels of tumor necrosis factor alpha (TNF-alpha), less GVHD histopathologic damage, and reduced donor T-cell expansion. Mechanistic analysis of this phenomenon demonstrated that dendritic cells (DCs) from gammadelta-/- recipients exhibited less allostimulatory capacity compared to wt DCs after irradiation. Normal DCs derived from BM caused greater allogeneic T-cell proliferation when cocultured with gammadelta T cells than DCs cocultured with medium alone. This enhancement did not depend on interferon gamma (IFN-gamma), TNF-alpha, or CD40 ligand but did depend on cell-to-cell contact. These data demonstrated that the host gammadelta T cells exacerbate GVHD by enhancing the allostimulatory capacity of host antigen-presenting cells. PMID- 15797997 TI - Apoptosis-based therapies for hematologic malignancies. AB - Apoptosis is an intrinsic cell death program that plays critical roles in tissue homeostasis, especially in organs where high rates of daily cell production are offset by rapid cell turnover. The hematopoietic system provides numerous examples attesting to the importance of cell death mechanisms for achieving homeostatic control. Much has been learned about the mechanisms of apoptosis of lymphoid and hematopoietic cells since the seminal observation in 1980 that glucocorticoids induce DNA fragmentation and apoptosis of thymocytes and the demonstration in 1990 that depriving colony-stimulating factors from factor dependent hematopoietic cells causes programmed cell death. From an understanding of the core components of the apoptosis machinery at the molecular and structural levels, many potential new therapies for leukemia and lymphoma are emerging. In this review, we introduce some of the drug discovery targets thus far identified within the core apoptotic machinery and describe some of the progress to date toward translating our growing knowledge about these targets into new therapies for cancer and leukemia. PMID- 15797999 TI - Hepcidin levels in humans are correlated with hepatic iron stores, hemoglobin levels, and hepatic function. AB - Hepcidin, a key regulator of iron metabolism, is synthesized by the liver. Hepcidin binds to the iron exporter ferroportin to regulate the release of iron into plasma from macrophages, hepatocytes, and enterocytes. We analyzed liver samples from patients undergoing hepatic surgery for cancer or receiving liver transplants and analyzed correlations between clinical parameters and liver hepcidin mRNA and urinary hepcidin concentrations. Despite the many potential confounding influences, urinary hepcidin concentrations significantly correlated with hepatic hepcidin mRNA concentrations, indicating that hepcidin quantification in urine is a valid approach to evaluate hepcidin expression. Moreover, we found in humans that hepcidin levels correlated with hepatic iron stores and hemoglobin levels and may also be affected by hepatic dysfunction. PMID- 15797998 TI - Internal tandem duplications of the FLT3 gene are present in leukemia stem cells. AB - Internal tandem duplication mutations of the FLT3 gene (FLT3/ITD mutations) are the most frequent molecular abnormality in acute myeloid leukemia (AML) and are associated with a poor overall survival. While the normal FLT3 receptor is expressed in early hematopoietic progenitor cells, it has not been determined whether FLT3 mutations are present in the leukemic stem cells. In this study, we sorted primary AML samples into stem cell-enriched CD34+/CD38- fractions and then analyzed the sorted and unsorted cells for the FLT3 mutant-wild-type ratio. In each case, the FLT3 mutant-wild-type ratio was not changed by selection of CD34+/CD38- cells, implying that the mutations are present in the leukemic stem cells. We used the stem cell-enriched fraction to engraft nonobese diabetic severe combined immunodeficient (NOD-SCID) mice and then confirmed that the FLT3/ITD mutation was present in the resultant engrafted marrow. As a final test of the importance of FLT3/ITD signaling in this engraftment model, we used a small molecule FLT3 inhibitor, CEP-701, to inhibit engraftment of FLT3/ITD stem cells. Taken together, these experiments establish that the FLT3/ITD mutations are present in leukemia stem cells, and that FLT3 inhibitors may have activity against these cells. PMID- 15798000 TI - Human erythropoietin gene therapy for patients with chronic renal failure. AB - Gene therapy holds a major promise. However, until now, this promise was fulfilled only in few cases, in rare genetic diseases. One very common clinical condition is anemia. Patients with anemia of chronic renal failure are treated with erythropoietin. The objective of this study was to develop a therapeutic platform for serum-secreted proteins like erythropoietin. We developed a tissue protein factory based on dermal cores (Biopump) harvested and implanted autologously. In this study, an adenovector was designed to express the human erythropoietin under the control of the cytomegalovirus (CMV) promoter. This vector transduced the harvested dermal cores ex vivo. The transduced cores were implanted, and erythropoietin and reticulocyte counts were measured. Dermal cores were harvested from 13 patients with chronic renal failure, and implantation was performed in 10. There were no significant drug-related side effects to this procedure. Erythropoietin serum levels increased significantly to therapeutic levels from day 1 after implantation reaching a peak during the first week of follow-up. The expression period was transient for up to 14 days. The rise of erythropoietin was followed by a transient significant increase in reticulocyte counts. The decrease of erythropoietin expression coincided with a significant dermal infiltrate of CD8 cytotoxic T cells. Antierythropoietin antibodies were not detected until day 90 following implantation. Implantation of dermal cores ex vivo transduced with human genes could eventually be used in the clinical setting to express therapeutic serum proteins. However, nonimmunogenic delivery system should be tested as gene vehicles. PMID- 15798001 TI - Inhibitory effect of the 4-aminotetrahydroquinoline derivatives, selective chemoattractant receptor-homologous molecule expressed on T helper 2 cell antagonists, on eosinophil migration induced by prostaglandin D2. AB - Prostaglandin (PG) D2, a major cyclooxygenase metabolite generated from immunologically stimulated mast cells, is known to induce activation and chemotaxis in eosinophils, basophils, and T helper 2 (Th2) lymphocytes via a newly identified PGD2 receptor, chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2). CRTH2 is hypothesized to play an important role in the outcome of allergic responses. However, the absence of selective CRTH2 antagonists has prevented the elucidation of the role of CRTH2 in pathogenesis of allergic diseases. We now report compounds discovered as selective CRTH2 antagonists, (2R*,4S*)-N-(1-benzoyl-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)-N phenylisobutyramide (K117) and (2R*,4S*)-N-(1-benzoyl-2-methyl-1,2,3,4 tetrahydroquinolin-4-yl)-N-phenylcyclopropanecarboxamide (K604). K117 and K604 have inhibitory effects on human CRTH2 with Ki values of 5.5 and 11 nM, respectively. The effect of these compounds is CRTH2-specific with no cross reactivity against 15 other receptors and four arachidonic acid-metabolizing enzymes. K117 and K604 has no effect on the basal Ca2+ level and inhibited the Ca2+ response induced by PGD2 in 293EBNA cells expressing human CRTH2. Also, K117 and K604 inhibit PGD2-induced human eosinophil chemotaxis with IC50 values of 7.8 and 42.2 nM, respectively, but they do not inhibit the CC-chemokine receptor 3 agonist eotaxin-induced chemotaxis. These results indicate that K117 and K604 are highly potent and selective antagonists for human CRTH2. These compounds have possibilities to become useful tools to explore CRTH2 functions in allergic diseases. PMID- 15798002 TI - Activation of peroxisome proliferator-activated receptor alpha by substituted urea-derived soluble epoxide hydrolase inhibitors. AB - Soluble epoxide hydrolase (sEH) plays a major role in regulating vascular epoxyeicosatrienoic acid metabolism and function, and substituted urea derivatives that inhibit sEH activity reduce blood pressure in hypertensive rats. We found that substituted urea derivatives containing a dodecanoic acid group, besides effectively inhibiting sEH, increased peroxisome proliferator-activated receptor (PPAR) alpha activity. In PPARalpha transfected COS-7 cells, treatment with 10 microM N-cyclohexyl-N'-dodecanoic acid urea (CUDA) or N-adamantanyl-N' dodecanoic acid urea (AUDA) produced 6- and 3-fold increases, respectively, in PPARalpha activation. Neither CUDA nor AUDA activated PPARdelta or PPARgamma directly, indicating selectivity for PPARalpha. CUDA did not alter PPARalpha protein expression, and it competitively inhibited the binding of Wy-14643 (pirinixic acid) to the ligand binding domain of PPARalpha, suggesting that it functions as a PPARalpha ligand. CUDA and AUDA were metabolized to chain shortened beta-oxidation products, a process that reduced their potency as sEH inhibitors and their ability to bind and activate PPARalpha. N,N' Dicylclohexylurea and N-cyclohexyl-N'-dodecylurea, sEH inhibitors that do not contain a carboxylic acid group, did not activate PPARalpha. In HepG2 cells, CUDA increased the expression of the PPARalpha-responsive gene carnitine palmitoyltransferase 1A. We conclude that CUDA and AUDA, by virtue of their carboxylic acid substitution, activate PPARalpha in addition to potently inhibiting sEH. Further development of these compounds could lead to a class of agents with hypotensive and lipid-lowering properties that may be valuable for the prevention and treatment of cardiovascular disease. PMID- 15798004 TI - Stress impairs murine intestinal barrier function: improvement by glucagon-like peptide-2. AB - Stress-induced intestinal barrier dysfunction may be involved in chronic intestinal disorders. Glucagon-like peptide-2 (GLP-2) is an intestinotrophic growth hormone that can rapidly improve intestinal epithelial barrier function. Here, we investigated whether mouse intestine is responsive to chronic psychological stress and whether pretreatment with GLP-2 can ameliorate stress induced changes. Mice were subjected to water avoidance stress (WAS; 1 h/day for 10 days) with GLP-2 or saline administered 4 h before each WAS session. After the final stress period, the intestine was removed for assessment of physiological/morphological changes. Compared with controls (sham-stressed mice), stressed mice demonstrated enhanced ion secretion and permeability in the jejunum, ileum, and colon. In addition, increased numbers of bacteria were observed adhering to and/or penetrating the epithelium, associated with infiltration of mononuclear cells into the mucosa. GLP-2 treatment improved intestinal barrier function in stressed mice and ameliorated other aspects of impaired host defense. Our study extends previous findings in rats of stress induced intestinal dysfunction and provides insights into potential novel therapeutics. PMID- 15798003 TI - Protein kinase C epsilon regulates tumor necrosis factor-alpha-induced stannin gene expression. AB - Stannin (Snn) is a highly conserved vertebrate protein that has been closely linked to trimethyltin (TMT) toxicity. We have previously demonstrated that Snn is required for TMT-induced cell death. Others have shown that TMT exposure results in tumor necrosis factor-alpha (TNFalpha) production and that TNFalpha treatment induces Snn gene expression in human umbilical vein endothelial cells (HUVECs). In this study, we investigated a signaling mechanism by which Snn gene expression is regulated by TMT and demonstrated that TNFalpha stimulates Snn gene expression in a protein kinase C epsilon-dependent manner in HUVECs in response to TMT exposure. Supporting this, we show that TMT-induced toxicity is significantly blocked by pretreatment with an anti-TNFalpha antibody in HUVECs. Using a quantitative real-time polymerase chain reaction assay, we also show that the level of Snn gene expression is significantly increased in HUVECs in response to either TMT or TNFalpha treatment. This TNFalpha-induced Snn gene expression is blocked when HUVECs were pretreated with bisindolylmaleimide I, an inhibitor of protein kinase C (PKC). In contrast, when HUVECs were treated with phorbol 12 myristate 13-acetate, a PKC activator, we observed a significant increase in Snn gene expression. Using isotype-specific siRNA against PKC, we further show that knockdown of PKC epsilon, but not PKC delta or PKC zeta, significantly blocked TNFalpha-induced Snn gene expression. Together, these results indicate that TNFalpha-induced, PKC epsilon-dependent Snn expression may be a critical factor in TMT-induced cytotoxicity. PMID- 15798006 TI - Care neglect, supervisory neglect, and harsh parenting in the development of children's aggression: a replication and extension. AB - To understand the effects of neglectful parenting, poor supervision, and punitive parenting in the development of children's aggression, 218 children ages 4 to 8 years who were disadvantaged and their mothers were recruited from two states to develop a sample that was diverse with respect to degree of urbanization and ethnicity. Multimethod and multisource indices of the predictive constructs (Social Disadvantage, Denial of Care Neglect, Supervisory Neglect, and Punitive Discipline) and the criterion construct (Aggression) were used in a test of a theoretical model using structural equation modeling. The results established the role of care neglect, supervisory neglect, and punitive parenting as mediators of the role of social disadvantage in the development of children's aggression, the importance of distinguishing between two subtypes of neglect, and the need to consider the role of discipline in concert with neglect when attempting to understand the parenting in the development of aggression. PMID- 15798005 TI - A novel neurotrophic agent, T-817MA [1-{3-[2-(1-benzothiophen-5-yl) ethoxy] propyl}-3-azetidinol maleate], attenuates amyloid-beta-induced neurotoxicity and promotes neurite outgrowth in rat cultured central nervous system neurons. AB - Progressive neuronal loss in Alzheimer's disease (AD) is considered to be a consequence of the neurotoxic properties of amyloid-beta peptides (A beta). T 817MA (1-{3-[2-(1-benzothiophen-5-yl) ethoxy] propyl}-3-azetidinol maleate) was screened as a candidate therapeutic agent for the treatment of AD based on its neuroprotective potency against A beta-induced neurotoxicity and its effect of enhancing axonal regeneration in the sciatic nerve axotomy model. The neuroprotective effect of T-817MA against A beta(1-42) or oxidative stress induced neurotoxicity was assessed using a coculture of rat cortical neurons with glia. T-817MA (0.1 and 1 microM) was strongly protective against A beta(1-42) induced (10 microM for 48 h) or H2O2-induced (100 microM for 24 h) neuronal death. T-817MA suppressed the decrease of GSH levels induced by H2O2 exposure (30 microM for 4 h) in cortical neuron culture; therefore, T-817MA was likely to alleviate oxidative stress. Besides the neuroprotective effect, T-817MA (0.1 and 1 microM) promoted neurite outgrowth in hippocampal slice cultures and reaggregation culture of rat cortical neurons. T-817MA also increased the growth associated protein 43 content in the reaggregation culture of cortical neurons. These findings suggest that T-817MA exerts neuroprotective effect and promotes neurite outgrowth in rat primary cultured neurons. Based on these neurotrophic features, T-817MA may have a potential for disease modification and be useful for patients with neurodegenerative diseases, such as AD. PMID- 15798007 TI - Family connections: a program for preventing child neglect. AB - Family Connections was a demonstration program specifically designed to prevent child neglect. This article describes the development of prevention strategies and the assessment of outcomes for families who received two versions of the intervention. The sample included 154 families (473 children) in a poor, urban neighborhood who met risk criteria for child neglect and who were randomly assigned to receive either a 3- or 9-month intervention. Self-report and observational data were analyzed using analyses of variance (ANOVA) with repeated measures. Results for the entire sample indicated positive changes in protective factors (parenting attitudes, parenting competence, social support); diminished risk factors (parental depressive symptoms, parenting stress, life stress); and improved child safety (physical and psychological care of children) and behavior (decreased externalizing and internalizing behavior). Results further reflected no advantage of the 9-month intervention for improving parenting adequacy. Further testing of the intervention with other target populations is being conducted. PMID- 15798008 TI - Neglectful behavior by parents in the life history of university students in 17 countries and its relation to violence against dating partners. AB - This article reports the prevalence of neglectful behavior by parents of university students in 17 nations (6 in Europe, 2 in North America, 2 in Latin America, 5 in Asia, Australia, and New Zealand) and tests the hypothesis that neglect is a risk factor for violence against a dating partner. The percentage at each university who experienced neglectful behavior ranged from 3.2% to 36% (median 12%), and the percentage who perpetrated violence against dating partners ranged from 15% to 45% (median 28%). Multilevel modeling found that the more neglectful behavior experienced as a child the greater the probability of assaulting and injuring a dating partner and that the link between experiencing neglect and perpetrating violence is stronger at universities in which dating violence is more prevalent. Efforts to help parents avoid neglectful behavior can make an important contribution to primary prevention of partner violence and probably also other forms of child maltreatment. PMID- 15798009 TI - The role of fathers in risk for physical child abuse and neglect: possible pathways and unanswered questions. AB - Despite overrepresentation of fathers as perpetrators in cases of severe physical child abuse and neglect, the role they play in shaping risk for physical child abuse and neglect is not yet well understood. This article reviews the possible father pathways that may contribute to physical child abuse and neglect risk and their existing empirical support. The present empirical base implicates a set of sociodemographic factors in physical maltreatment risk, including fathers' absence, age, employment status, and income they provide to the family. As well, paternal psychosocial factors implicated in physical child maltreatment risk include fathers' abuse of substances, their own childhood experiences of maltreatment, the nature of fathers' relationships with mothers, and the direct care they provide to the child. However, the empirical base presently suffers from significant methodological limitations, preventing more definitive identification of risk factors or causal processes. Given this, the present article offers questions and recommendations for future research and prevention. PMID- 15798010 TI - The psychobiology of neglect. AB - Child neglect, the most prevalent form of child maltreatment, is associated with adverse psychological and educational outcomes. It is hypothesized that these outcomes may be caused by adverse brain development. However, there are very few published cross-sectional studies and no prospective studies that examine the neurodevelopmental consequences of neglect. In this article, the author comprehensively outlines the issues involved in the psychobiological research of child neglect. Pre-clinical and clinical studies will be reviewed. Throughout the article, suggestions for future research opportunities and novel ways to address methodological difficulties inherent in this field of study are offered. The results of recent neuroimaging studies of maltreated children may provide a basis for understanding the early effects of neglect on childhood brain development. The author is comprehensively examining these issues as part of the Federal Child Neglect Consortium. PMID- 15798012 TI - Toward a definition of neglect in young children. AB - This study examined the relationship between child experiences identified conceptually as "neglectful" prior to age 4 and child outcomes at age 4. This was done using measures from two sites collected as part of LONGSCAN. Child needs were included within categories of physical and psychological safety and security. Problems with residence safety or cleanliness and untreated behavioral problems predicted child impairments in language. CPS reports of failure to provide shelter predicted impairments in several developmental outcomes. A stimulating home environment predicted less impairment in cognitive development. Multiple changes in residence predicted externalizing behavior problems. Exposure to verbally aggressive discipline predicted more behavioral problems overall. Conversely, some indicators (such as caregiver transitions and lack of medical care) predicted less developmental impairment or fewer behavior problems in certain domains. The approach supports a conceptualization of neglect based on child developmental needs. Implications for practice and future research are discussed. PMID- 15798011 TI - Examination of a conceptual model of child neglect. AB - This study attempted to provide empirical support for conceptual definitions of child neglect. We identified 12 types of needs, conceptualizing neglect as occurring when children's basic needs are not adequately met. We examined measures administered to 377 children and caregivers at ages 4 and 6 years participating in longitudinal studies on child mal-treatment to identify potential indicators of these needs. Indicators were found for latent constructs, operationalizing three of the basic needs (emotional support and/or affection, protection from family conflict and/or violence, and from community violence). These latent constructs were used in a measurement model; this supported the conceptual definitions of neglect. A structural equation model then assessed whether the latent constructs were associated with child adjustment at age 8 years. Low level of perceived support from mother was associated with internalizing and externalizing behavior problems. Exposure to family conflict was also linked to these problems, and to social difficulties. Finally, children's sense of experiencing little early affection was associated with subsequent externalizing behavior and social problems. The approach of conceptualizing neglect in terms of unmet child needs, developing a measurement model to define latent neglect constructs, and relating these constructs to subsequent adjustment can build our understanding of neglect. PMID- 15798013 TI - Molecular interactions of the aryl hydrocarbon receptor and its biological and toxicological relevance for reproduction. AB - The dioxin/aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor responsive to both natural and man-made environmental compounds. AhR and its nuclear partner ARNT are expressed in the female reproductive tract in a variety of species and several indications suggest that the AhR might play a pivotal role in the physiology of reproduction. Furthermore, it appears to be the mediator of most, if not all, the adverse effects on reproduction of a group of highly potent environmental pollutants collectively called aryl hydrocarbons (AHs), including the highly toxic compound 2,3,7,8-tetrachlor-odibenzo-p-dioxin (TCDD). Although a large body of recent literature has implicated AhR in multiple signal transduction pathways, the mechanisms of action resulting in a wide spectrum of effects on female reproduction are largely unknown. Here we summarize the major types of molecular cross-talks that have been identified for the AhR and linked cell signaling pathways and that are relevant for the understanding of the role of this transcription factor in female reproduction. PMID- 15798014 TI - Bioscience-bioethics and life factors affecting reproduction with special reference to the Indigenous Australian population. AB - The demand for equality of recognition or respect is the dominant passion of modernity. The 20th century experienced a giant leap in technological inventiveness and ruthless use of technological power. In the 21st century, human welfare and environmental wellbeing demand fundamental political appraisal. We have the means, if we choose, to eradicate poverty and to responsibly protect the global environment. However, economic, political and cultural systems act to differentially allocate the benefits and risks for growth between socioeconomic groups. For example, it is a matter of pride that the neonatal mortality rate in affluent societies has dropped substantially since the late 1970s. However, the level of infant mortality (three times the national average) and low birthweight (13%) among the Indigenous Australian population is the highest in the country. With hindsight we now know that is the inevitable legacy of Australia's colonial history. Chronic physical and psychological stress is recognized as an important etiological factor in many lifestyle diseases of the cardiovascular, immune and reproductive systems. Diseases of adaptation are further advanced by non-adaptive lifestyle choices, depression, alcoholism and other drug dependencies. This review describes the principles of bioscience ethics and targets equity issues as they affect human reproduction across generations with particular reference to the Indigenous population of Australia. The review also considers ways we may advance global and cultural maturity from the Indigenous Australian perspective and proposes an ecologically based model of preventative care. If we are to embrace fundamental social change and protect future children without threatening parents' basic freedoms, then new beliefs and priorities--based on a compassionate understanding of biological systems--must evolve from the general public. Belief in human rights arising from a sense of human dignity is a collective outcome originating from individual commitment. The golden rule; that is, Nature's principle of reciprocity, is fundamental in bridging the gap between knowledge and effective action. PMID- 15798015 TI - Inducible nitric oxide synthase-derived nitric oxide regulates germinal vesicle breakdown and first polar body emission in the mouse oocyte. AB - The present study investigated the subcellular localization of inducible nitric oxide synthase (iNOS) during mouse oocyte meiotic maturation and fertilization using confocal microscopy, and further studied the roles of iNOS-derived NO in oocyte maturation by using an iNOS-specific inhibitor aminoguanidine (AG) and iNOS antibody microinjection. In germinal vesicle-stage oocytes, iNOS immunoreactivity was mainly localized in the germinal vesicle. Shortly after germinal vesicle breakdown, the iNOS immunoreactivity accumulated around the condensed chromosomes. At metaphase I and metaphase II, with the organization of chromosomes to the equatorial plate, iNOS immunoreactivity was concentrated around the aligned chromosomes, putatively the position of the metaphase spindle. The accumulation of iNOS immunoreactivity could not be detected at anaphase I and anaphase II. However, at telophase I and telophase II, the staining of iNOS was concentrated in the region between the separating chromosomes/chromatids. Furthermore, the staining of iNOS also accumulated in the male and female pronuclei in fertilized eggs. Germinal vesicle breakdown and the first polar body emission of the oocytes were significantly blocked by the iNOS-specific inhibitor AG in a dose-dependent manner. The germinal vesicle breakdown in oocytes injected with iNOS antibody was also inhibited. We found that the phosphorylation of mitogen-activated protein kinase in oocytes after germinal vesicle breakdown was inhibited by AG treatment. The control oocytes extruded a normal first polar body, while the AG-treated oocytes exhibited an elongated protrusion or no elongated protrusion. The results of confocal microscopy showed that the AG treated oocytes were arrested at anaphase I-telophase I. Our results suggest that the iNOS-derived NO pathway plays important roles in mouse oocyte meiotic maturation, especially in germinal vesicle breakdown and the anaphase-telophase transition. PMID- 15798016 TI - Oocytes recovered from cows treated with retinol become unviable as blastocysts produced in vitro. AB - Retinoids have been shown to enhance developmental competence of the oocyte in cattle, sheep and pigs. In this study we investigated whether exogenous retinol stimulates the bovine oocyte during its intrafollicular growth and the time limits of exposure to exogenous retinol. In addition, we also determined the efficiency of ovum pick-up techniques in combination with retinol treatment and the viability of embryos after transfer to recipients. In Experiment 1, heifers were injected with retinol or vehicle, and concentrations of retinol in the blood were analysed on Day 0 (prior to injection), Day 1 and, together with follicular fluid, Day 4. Blood retinol increased by Day 1 and cleared on Day 4, but retinol remained higher within the follicle. In Experiment 2, oocyte donors were injected weekly with retinol or vehicle four times during a twice-per-week cycle of eight recovery sessions (starting 4 days before the first session), followed by a second eight-session cycle without treatment. Oocytes recovered were fertilized and cultured in vitro. Retinol treatment yielded higher numbers of low-quality oocytes throughout, although retinol measured during cycles did not change. Total oocytes, and morulae and blastocyst rates, increased during the first five sessions following treatment with retinol. As previously shown with oocytes from slaughterhouse ovaries, retinoic acid stimulated blastocyst development. Following transfer to recipients, blastocysts from oocytes exposed to retinol were unable to establish pregnancy. Our study confirms the existence of an effect of retinol on the intrafollicular oocyte in the cow and provides evidence regarding the teratogenic effect of retinol. PMID- 15798017 TI - Brain-derived neurotrophic factor promotes bovine oocyte cytoplasmic competence for embryo development. AB - The ability of an oocyte to support early embryonic development requires both nuclear and cytoplasmic maturation. We have investigated the effects of brain derived neurotrophic factor (BDNF) on maturation of the bovine oocyte and embryo development after parthenogenetic activation. By RT-PCR and immunohistochemistry, cumulus and oocytes were shown to express mRNA and protein for BDNF and the p75 common neurotrophin receptor. However, mRNA for the BDNF-specific full length and truncated isoforms of the TrkB receptor are only detected in cumulus, suggesting that oocytes and cumulus differ in their capacity to respond to neurotrophin signalling. In in vitro maturation experiments, the proportion of cumulus oocyte complexes maturing to metaphase II was not altered by BDNF in groups lacking fetal calf serum (FCS), but was significantly lower than the positive control containing 10% FCS (P < 0.01). However, after maturation, the proportion of parthenogenetically activated oocytes forming blastocysts was highest for 10 ng/ml BDNF (24%, n = 95) followed by 100 ng/ml BDNF (18%, n = 91) and 10% FCS (15%, n = 103), which in turn were greater than no serum (10%, n = 83; P < 0.01). Maturation in the presence of a BDNF blocking antibody resulted in a blastocyst yield that was comparable to the absence of serum, and lower than in the presence of BDNF (P < 0.01). Similar effects on progression to metaphase II and blastocyst formation were observed using oocytes matured without cumulus. Together, these results provide the first evidence for a role for neurotrophins in promoting oocyte cytoplasmic competence to support embryonic development, despite being insufficient in the absence of serum to enhance nuclear maturation. PMID- 15798019 TI - Porcine embryo development and fragmentation and their relation to apoptotic markers: a cinematographic and confocal laser scanning microscopic study. AB - Porcine embryo selection prior to transfer is mainly influenced by morphological criteria. However, the relationship between embryonic morphology, developmental potential and cell death by apoptosis in porcine embryos is still unclear. The aim of this study was to establish embryo quality parameters for in vivo fertilised porcine embryos based on timing of development in vitro, embryo morphology and the presence of apoptosis. The kinetics of development and morphological parameters were investigated in a time-lapse cinematographic experiment. Possible links between embryo morphology and apoptosis were examined via a confocal laser scanning experiment, analysing nuclear changes, annexin V and terminal dUTP nick-end labelling. The timing of early cleavages was firmly linked to embryo developmental competence in vitro. Attainment of at least the 5 cell stage before 77 h post insemination and attainment of the morula stage before 102 h post insemination significantly increased the odds for reaching the early blastocyst stage. Overall, a negative effect of fragmentation percentage and fragmentation pattern on subsequent embryonic development was observed, but the developmental potential of embryos experiencing slight fragmentation (0-5%) was not different from embryos without fragmentation. Correlations detected between developmental arrest and fragmentation, and fragmentation and apoptosis were 0.60 and 0.87 (P < 0.05) respectively. Only a minority of the embryos arrested between the 1- and 4-cell stage displayed biochemical characteristics of apoptosis. Consequently, a significant correlation (0.57) between developmental arrest and apoptosis could only be established for embryos arrested after embryonic genome activation. PMID- 15798018 TI - The expression of calpain 1 and calpain 2 in spermatogenic cells and spermatozoa of the mouse. AB - There is some evidence suggesting that Ca2+ is involved in processes that occur during the development and function of spermatozoa. Calcium-dependent proteins, such as calmodulin, are expressed during mammalian spermatogenesis further suggesting that Ca2+ takes part in its regulation. However, the precise roles of Ca2+ in spermatogenesis remain to be elucidated. Calpains are a family of Ca(2+) dependent cysteine proteases whose members are expressed ubiquitously or in a tissue-specific manner. Calpain has been demonstrated to mediate specific Ca(2+) dependent processes including cell fusion, mitosis and meiosis. We herein followed the expression pattern of calpain's ubiquitous isoforms, 1 and 2, throughout spermatogenesis at the RNA and protein levels by RT-PCR and Western blotting analysis. Both RNA and protein studies revealed that these isoforms are expressed in all spermatogenic cells. The expression of calpain 1 levels is slightly higher in spermatocytes entering the meiotic phase. Both calpain isoforms are also expressed in mouse spermatozoa and are localized to the acrosomal cap. Inducing capacitated spermatozoa to undergo the acrosome reaction in the presence of a selective calpain inhibitor significantly reduced the acrosome reaction rate in a dose-dependent manner. Thus, calpain, a pluripotential protease with numerous substrates, may serve as an effector in more than one pathway in the complex process of spermatogenesis and in the events preceding fertilization, such as the acrosome reaction. PMID- 15798020 TI - Gene expression for LH receptor, 17 alpha-hydroxylase and StAR in the theca interna of preantral and early antral follicles in the bovine ovary. AB - The onset of gene expression for three proteins that play pivotal roles in theca interna function, namely the LH receptor (LH-R), cytochrome P450 17 alpha hydroxylase (17 alpha OH) and the steroidogenic acute regulatory protein (StAR), was determined. Ovaries were obtained on day 9 of the oestrus cycle from mature synchronized dairy cows (n = 5) and gene expression in preantral and antral follicles up to 4 mm in diameter was evaluated by in situ hybridization. LH-R and 17 alpha OH mRNAs were observed first, in the theca interna of large preantral follicles (type 4), concurrent with its morphological differentiation. StAR mRNA appeared later during follicular growth, in follicles >1 mm in diameter (type 6). LH-R and 17 alpha OH mRNAs were found exclusively in the thecal cells, whereas StAR mRNA appeared in thecal cells, granulosa cells of late atretic follicles and oocytes. In early atresia, thecal cells expressed all three mRNAs, and their expression decreased gradually as atresia progressed. Atresia in granulosa cells was characterized by massive apoptosis of periantral, but not peribasal cells, that differentiated into luteal-like cells expressing StAR. In summary, our study suggests that in spite of the presence of 17 alpha OH, a key enzyme in steroidogenesis, the ability to produce steroids by bovine follicles smaller than 1 mm in diameter must be very limited due to the absence of StAR protein. During the early stages of atresia, thecal cells remain morphologically and functionally healthy, and continue to express all three studied mRNAs. PMID- 15798021 TI - Heat stress diminishes gonadotropin receptor expression and enhances susceptibility to apoptosis of rat granulosa cells. AB - Heat stress inhibits ovarian follicular development in mammalian species. We hypothesized that heat stress inhibits the function of follicular granulosa cells and suppresses follicular development. To test this, immature female rats were injected with pregnant mare serum gonadotropin (PMSG) at 48 h after the start of temperature treatment (control: 25 degrees C, 50% RH; heat stress: 35 degrees C, 70% Relative Humidity). The ovaries and granulosa cells of follicles at different developmental stages were analyzed for gonadotropin receptor levels and aromatase activity; estradiol levels were measured in follicular fluid. Before injection, heat stress diminished only the amount of FSH receptor on granulosa cells of antral follicles. During PMSG-stimulated follicular development, heat stress strongly inhibited gonadotropin receptor levels and aromatase activity in granulosa cells, and estradiol levels in the follicular fluid of early antral, antral and preovulatory follicles. To examine apoptosis and mRNA levels of bcl-2 and bax in granulosa cells, follicles harvested 48 h after PMSG injection were cultured in serum-free conditions. Heat-stressed granulosa cells showed a time dependent increase in apoptosis. The bcl-2 mRNA levels were similar in control and heat-stressed granulosa cells; bax mRNA levels were increased in heat stressed granulosa cells. According to these results, heat stress inhibits expression of gonadotropin receptors in granulosa cells and attenuates estrogenic activity of growing follicles, granulosa cells of heat-stressed follicles are susceptible to apoptosis, and the bcl2/bax system is not associated with heat stress-induced apoptosis of granulosa cells. Our study suggests that decreased numbers and function of granulosa cells may cause ovarian dysfunction in domestic animals in summer. PMID- 15798022 TI - Bone morphogenetic protein 15 and growth differentiation factor 9 co-operate to regulate granulosa cell function. AB - The oocyte-secreted polypeptide growth factors, growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15, also known as GDF9B) have both been shown to be essential for ovarian follicular growth and function. The effects of murine (m) and ovine (o) GDF9 as well as oBMP15, alone or together, on 3H-thymidine uptake and progesterone and inhibin production by granulosa cells from rats were determined. Murine GDF9 stimulated thymidine incorporation by granulosa cells whereas oGDF9 and oBMP15 alone had no effect. However, oBMP15 given together with mGDF9 or oGDF9 was very potent in stimulating 3H-thymidine incorporation by granulosa cells with a greater than 3-fold stimulation compared with any growth factor alone. The synergistic effect of oBMP15 and oGDF9 was almost completely blocked by antibodies generated against these growth factors when administered either alone or in combination. While neither GDF9 (murine or ovine) nor oBMP15 were able to modulate FSH-stimulated progesterone production on their own, FSH-stimulated progesterone production by granulosa cells was potently inhibited when BMP15 and GDF9 were administered together. Immunoreactive alpha inhibin levels increased more than 15-fold from granulosa cells when BMP15 and GDF9 were given together whereas consistent stimulatory effects of either growth factor alone were not observed. The effects of GDF9 and BMP15, when added together, were different than those observed for the growth factors alone. Therefore, we hypothesize that within the ovary, these oocyte-secreted growth factors co-operate to regulate proliferation and gonadotropin-induced differentiation of granulosa cells in mammals. PMID- 15798023 TI - Bone morphogenetic protein 15 and growth differentiation factor 9 co-operate to regulate granulosa cell function in ruminants. AB - The oocyte-secreted polypeptide growth factors, growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15, also known as GDF9B) have both been shown to be essential for ovarian follicular development and ovulation rate. In addition, it is known from both in vivo and in vitro studies that these factors co-operate in some manner. To date, most studies examining the in vitro effects of these growth factors have used the rodent model. However, the evidence suggests that these growth factors have somewhat different roles between rodents and ruminants. Therefore, the objectives of these studies were to examine the effects of GDF9 and BMP15, alone and together, on the functions of ovine and bovine granulosa cells under in vitro conditions. Ovine (o)BMP15 given together with murine (m)GDF9 or oGDF9 was more potent in stimulating (3)H-thymidine incorporation by ovine granulosa cells compared with each growth factor alone. For bovine granulosa cells, there appeared to be little or no co-operativity between oBMP15 and oGDF9 as oBMP15 alone was as potent as any combination of the two growth factors in stimulating (3)H-thymidine uptake. The species of origin of GDF9 affected the progesterone response in ovine granulosa cells with mGDF9 stimulating and oGDF9 inhibiting progesterone production. Ovine BMP15 alone had no effect on progesterone production by ovine granulosa cells and these growth factors did not appear to co-operate. FSH-stimulated progesterone production by bovine granulosa cells was most potently inhibited when oBMP15 and murine or ovine GDF9 were administered together. As was observed for progesterone, the species of origin of GDF9 affected inhibin production by ovine granulosa cells where mGDF9 inhibited while oGDF9 stimulated production. Murine GDF9 also inhibited inhibin production from bovine granulosa cells. For both ovine and bovine granulosa cells, BMP15 alone had no effect on inhibin production and there did not appear to be any co-operation between GDF9 and BMP15. These results indicate that the effects of BMP15 and GDF9 varied with respect to the species of origin of the growth factor. Moreover, the effects of GDF9 and BMP15 together were often co-operative and not always the same as those observed for these growth factors alone. PMID- 15798024 TI - In vivo effects of pregnancy-associated plasma protein-A, activin-A and vascular endothelial growth factor on other follicular-fluid factors during follicle deviation in mares. AB - During a follicular wave in mares, the two largest follicles (F1 and F2) begin to deviate in diameter when F1 is a mean of 22.5 mm. The intrafollicular effects of pregnancy-associated plasma protein-A (PAPP-A), IGF-I, activin-A and vascular endothelial growth factor (VEGF) on other follicular-fluid factors during deviation were studied. In four treated groups (n = 7/group), a single dose of one of the four factors was injected into F2 when F1 was > or = 20.0 mm (expected beginning of deviation). In a control group (n = 7), F2 was injected with vehicle. One day after treatment, a sample of follicular fluid was taken from F1 and F2 of the control group and from F2 of the treated groups and was assayed for free IGF-I, oestradiol, androstenedione, activin-A, inhibin-A, follistatin and VEGF. In the control group, the means for all end points were significantly greater in F1 than in F2, except that concentrations of androstenedione were lower in F1 than in F2. The treatment effects for F2 were significant as follows: PAPP-A increased the concentrations of free IGF-I, inhibin-A, follistatin and VEGF and decreased the concentrations of androstenedione; IGF-I increased the concentration of inhibin-A and decreased the concentration of androstenedione; activin-A decreased the concentrations of follistatin and androstenedione and increased the diameter of F2; and VEGF increased the concentration of IGF-I and decreased the concentration of androstenedione. These results support the hypotheses that during deviation in mares PAPP-A increases the follicular-fluid concentrations of free IGF-I, follistatin responds to changes in follicular-fluid concentrations of activin-A, and VEGF affects the concentrations of other follicular-fluid factors. PMID- 15798025 TI - Effect of steroids and nitric oxide on pituitary hormone release in ovariectomized, peripubertal rats. AB - The purpose of this study was to determine the effects of the duration of steroid depletion on the steroid-induced luteinizing hormone and prolactin surges in ovariectomized, peripubertal female rats. Additionally, the role of nitric oxide (NO) in mediating the surge responses was determined. Peripubertal, 6-week-old, female Sprague-Dawley rats were ovariectomized. One or three weeks later, animals were injected with 17 beta-estradiol (50 microg, sc) followed 48 h later by progesterone (2.5 mg, sc). Effects of NO were examined by administering L arginine (300 mg/kg, ip). The response of ovariectomized, adult females to steroid treatment was also determined. One and three weeks after ovariectomy, steroid replacement produced an LH and prolactin surge in peripubertal animals. However, both the magnitude and duration of the LH surge was greater 3 weeks after ovariectomy. While L-arginine significantly enhanced the magnitude of the LH surge 1 week after ovariectomy, by 3 weeks L-arginine caused a decrease in the duration, but not the magnitude of the surge. In contrast, L-arginine did not affect either the magnitude or duration of the prolactin surge one week after ovariectomy, but diminished the magnitude after 3 weeks of steroid depletion. In adults, steroids induced significant increases in both LH and prolactin. These results demonstrate that sensitivity to NO stimulation of LH, but not prolactin secretion, is modulated by the duration of gonadal steroid hormone depletion. The differences in the responsiveness of LH and prolactin to steroid-induced stimulation in peripubertal animals demonstrate that these hormones are regulated by NO through different mechanisms. PMID- 15798026 TI - Impact of a mild scrotal heat stress on DNA integrity in murine spermatozoa. AB - An increase in scrotal temperature can lead to the production of poor quality spermatozoa and infertility. In the present study we have used mice to examine the impact of mild, scrotal heat stress (42 degrees C for 30 min) on numbers of spermatozoa as well as on the integrity of their DNA. Spermatozoa recovered from the epididymides hours (1 to 24) or days (7 to 32) after treatment were analysed using COMET and sperm chromatin structure (SCSA) assays. The treatment induced a stress response in both the testis and the epididymis that was associated with reduced expression of the cold inducible RNA binding protein (Cirp) and an increase in germ cell apoptosis (Apotag positive cells). Although spermatozoa present in the epididymis at the time of heating contained correctly packaged DNA, its integrity was compromised by heat stress. In addition, although some germ cells, which were present within the testis at the time of heat stress, were removed by apoptosis, many germ cells completed their development and were recovered as motile spermatozoa with damaged DNA. In conclusion, these data demonstrate that scrotal heat stress can compromise the DNA integrity of spermatozoa and this may have clinical implications for patients undergoing IVF and intra-cytoplasmic sperm injection (ICSI). PMID- 15798027 TI - The PATE gene is expressed in the accessory tissues of the human male genital tract and encodes a secreted sperm-associated protein. AB - The PATE gene is expressed in prostate and testis. To determine if PATE is expressed in other accessory tissues of the male genital tract, RT-PCR of the epididymis and seminal vesicle was performed. PATE mRNA was highly expressed in the epididymis and seminal vesicle. In situ hybridization of the testis showed PATE mRNA is strongly expressed in the spermatogonia. The PATE gene encodes a 14 kDa protein with a predicted signal sequence and a cleavage site between residues G21 and S22. To determine if PATE is a secreted protein, 293T cells were transfected with a pcDNA-PATE-myc-His plasmid and protein immunoprecipitated with anti-myc monoclonal antibody. Western blot analysis showed the presence of PATE myc-His protein was in the medium and the cell lysate. Confocal microscopy demonstrated that PATE-myc-His protein is found in the endoplasmic reticulum. The polyclonal antibody SOL-1 was generated by immunization of rabbits with recombinant PATE protein expressed and purified from Escherichia coli. Western blots were performed on extracts of prostate, testis, seminal vesicle and ejaculated spermatozoa, but PATE protein was only detected in the spermatozoa. Immunostaining of sperm smears revealed that PATE is located in a band-like pattern in the sperm head. Our data indicate that PATE is made by various sexual accessory tissues and secreted into the semen where it becomes associated with sperm, suggesting that PATE is a novel sperm-associated protein with a possible role in mammalian sperm maturation. PMID- 15798028 TI - Expression of TASK and TREK, two-pore domain K+ channels, in human myometrium. AB - Two-pore domain K+ channels are an emerging family of K+ channels that may contribute to setting membrane potential in both electrically excitable and non excitable cells and, as such, influence cellular function. The human uteroplacental unit contains both excitable (e.g. myometrial) and non-excitable cells, whose function depends upon the activity of K+ channels. We have therefore investigated the expression of two members of this family, TWIK (two-pore domain weak inward rectifying K+ channel)-related acid-sensitive K+ channel (TASK) and TWIK-related K+ channel (TREK) in human myometrium. Using RT-PCR the mRNA expression of TASK and TREK isoforms was examined in myometrial tissue from pregnant women. mRNAs encoding TASK1, 4 and 5 and TREK1 were detected whereas weak or no signals were observed for TASK2, TASK3 and TREK2. Western blotting for TASK1 gave two bands of approximately 44 and 65 kDa, whereas TREK1 gave bands of approximately 59 and 90 kDa in myometrium from pregnant women. TASK1 and TREK1 immunofluorescence was prominent in intracellular and plasmalemmal locations within myometrial cells. Therefore, we conclude that the human myometrium is a site of expression for the two-pore domain K+ channel proteins TASK1 and TREK1. PMID- 15798029 TI - Estradiol and leptin as conditional prognostic IVF markers. AB - We studied the concentration of serum estradiol and serum and follicular fluid leptin in 200 women undergoing their first in vitro fertilization with embryo transfer (IVF-ET) program at the time of human chorionic gonadotrophin administration and oocyte retrieval, in an attempt to assess their concerted role on embryo quality and the prognosis of IVF outcome. Low serum (46.49 +/- 8.4 ng/ml) and follicular fluid (52 +/- 9.8 ng/ml) leptin levels were associated with a high number of 'good-quality' embryos (73.6%) and high implantation (11.2%) and pregnancy (35.8%) rates and were observed in women with normal peak estradiol levels of between 1000 and 2000 pg/ml. It appears that leptin and estradiol interact coordinately in a concentration-dependent manner to control IVF outcome. Further studies will be required to substantiate and clarify the mechanism of proposed conditional interaction between the two hormonal systems. PMID- 15798030 TI - Modifications of bull spermatozoa induced by three extenders: Biociphos, low density lipoprotein and Triladyl, before, during and after freezing and thawing. AB - The success of artificial insemination with frozen semen implies the reduction of the deleterious effects on the cells induced by this technique. These effects can occur as early as during the first dilution in an extender, as well as at any step, during or after the freezing process. In this work, we have compared the modifications induced by Triladyl, low density lipoproteins (LDL) and Biociphos extenders, after dilution and cooling to 4 degrees C for 1, 4 and 24 h. Alterations in the cell structures were visualized by electron microscopy (EM). More than 80% of spermatozoa were injured after incubation for 4 h in Triladyl, while 3% and 47% were counted in LDL and Biociphos respectively. This latter extender was deleterious to cell membrane integrity after incubation for 4 h or longer. The ultrastructure of frozen spermatozoa was studied by EM of cryofixed cryosubstituted samples obtained from regular 0.5 ml French straws frozen using our usual protocol. The main differences between samples concerned the size and appearance of the frozen extender veins, while very few cell defects were found to be added by the freezing process at any depth in the straws. After thawing, semen motility was twofold higher (P < 0.05) in Biociphos (64%) and LDL (61%) than in Triladyl (32%) and the cells were less altered in LDL. We concluded that the LDL extender offers a better protection for storage of frozen spermatozoa, and can probably also be used for the preservation of fresh semen for short periods. PMID- 15798032 TI - Obstructive sleep apnoea syndrome: underestimated and undertreated. AB - The obstructive sleep apnoea syndrome (OSAS) was first identified only 40 years ago and its clinical importance is increasingly recognized. Although now acknowledged as a worldwide problem, which in Western countries affects 2-4% of middle-aged men and 1-2% of middle-aged women, the majority of affected individuals remain undiagnosed. OSAS is strongly associated with obesity but is also increasingly identified in the less obese, in whom a particular craniofacial structure is an important contributory factor. The prevalence of OSAS is likely to be increasing in parallel with the epidemic of obesity currently occurring in many countries. The common presenting complaints are excessive daytime sleepiness and loud snoring. The sleepiness impairs social functioning, work performance and driving ability, and accounts for a large socio-economic burden on the community. Hypertension is an important independent association. The treatment of choice is nocturnal continuous positive airway pressure which is highly effective and is also cost effective. PMID- 15798033 TI - Physicians, families, and population health. PMID- 15798034 TI - The family contribution to health status: a population-level estimate. AB - PURPOSE: Clinical studies have shown strong family influences on individual health, but the aggregate importance of family effects for population health is unknown. Our objective was to estimate, at a population level, the variance in individual health status attributable to the family. METHODS: Secondary data were used from the Community Tracking Study, a stratified random sample of the US population. Hierarchical linear modeling was used to estimate the individual and family components of health status. The setting was 60 US communities, which account for approximately one half of the population. Participants were US residents aged 18 years and older who shared a household with family members in the study (N = 35,055). Main outcome measures were the Short Form-12 (SF-12) self reported physical and mental subscales. RESULTS: Depending on the family configuration, 4.5% to 26.1% of the variance in individual health status was derived from the family. The proportion was highest for older married persons. The family effect on health status was generally similar for physical and mental health. Including age, family income, and health insurance status in the regression equations moderately reduced the family variance component. CONCLUSIONS: At a population level, the family contribution to individual health status is measurable and substantial. The shared characteristics of income and health insurance account for only a modest portion of the effect. Health policy and interventions should place more emphasis on the family's role in health. PMID- 15798035 TI - Long-term results from a randomized controlled trial to increase cancer screening among attendees of community health centers. AB - PURPOSE: We assessed whether increased cancer screening rates that were observed with Cancer Screening Office Systems (Cancer SOS) could be maintained at 24 months' follow-up, a period in which clinics were expected to be largely self sufficient in maintaining the intervention. METHODS: Eight primary care clinics serving disadvantaged populations in Hills-borough County, Fla, agreed to take part in a cluster-randomized experimental trial. Charts of independent samples of established patients aged 50 to 75 years were abstracted, with data collected at baseline (n = 1,196) and at 24 months' follow-up (n = 1,296). Papanicolaou (Pap) smears, mammography, and fecal occult blood testing were assessed. RESULTS: At 24 months of follow-up, intervention patients had received a greater number of cancer screening tests (mean 1.17 tests vs 0.94 tests, t test = 4.42, P <.0001). In multivariate analysis that controlled for baseline screening rates, secular trends, and other patient and clinic characteristics, the intervention increased the odds of mammograms slightly (odds ratio [OR]) = 1.26; 95% confidence interval [CI], 1.02-1.55; P = .03) but had no effect on fecal occult blood tests (OR = 1.17; 95% CI, 0.92-1.48; P =0.19) or Pap smears (OR = 0.88; 95% CI, 0.0.68-1.15; P = .34). CONCLUSIONS: The Cancer SOS intervention had persistent, although modest, effects on screening at 24 months' follow-up, an effect that had clearly diminished from results reported at 12 months' follow-up. Further study is needed to develop successful intervention strategies that are either self-sustaining or that are able to produce long-term changes in screening behavior. PMID- 15798038 TI - Office-based unsedated ultrathin esophagoscopy in a primary care setting. AB - PURPOSE: Gastroesophageal reflux disease is common and with time may be complicated by Barrett's esophagus and esophageal adenocarcinoma. Upper gastrointestinal endoscopy, including esophagoscopy, is the procedure of choice to diagnose Barrett's esophagus and other esophageal disease. The use of unsedated ultrathin esophagoscopy (UUE) has been reported by gastroenterologists in specialized endoscopy units and otolaryngologists in outpatient otolaryngology offices, but UUE has not been previously described in a primary care setting. This study examines the feasibility of office-based UUE in primary care. METHODS: This study is a retrospective chart review in a university-based family medicine clinic in the southeastern United States. Charts were reviewed of 56 adult outpatients who were referred for further evaluation of reflux symptoms that persisted after at least 4 weeks of therapy with histamine(2) receptor agonists or proton pump inhibitors and who elected to undergo UUE in the primary care setting. Patient demographics, procedure indications and findings, changes in clinical management, and procedure times were recorded. RESULTS: One hundred percent of patients asked to participate in UUE were willing to undergo the procedure (mean age 48.3 +/- 1.6 y, 57.1% women); 95% of the patients tolerated UUE. Barrett's esophagus was diagnosed in 5.7% (n = 3) of the patients. Mean procedure time was 5.5 +/- 1.7 min. No complications were reported in this series. CONCLUSIONS: Initial data suggest that UUE is feasible in primary care, with the majority of patients tolerating the procedure. UUE may be an efficient method of examining the distal esophagus. PMID- 15798036 TI - Liability implications of physician-directed care coordination. AB - PURPOSE: Various public and private initiatives encourage physicians to coordinate care for patients who have multiple chronic conditions, but physicians may resist doing so for fear of liability. This article assesses the extent of liability risk. METHODS: This qualitative study combines legal research with key informant interviews. Relevant legal authorities were identified through literature searches in legal databases. In-depth interviews were conducted with a purposive sample of 16 key informants who have relevant expertise in malpractice insurance, risk management, or liability law. Additionally, 19 other participants with relevant experience in case management or managed care were briefly queried about their liability experience relating to care management. RESULTS: Some aspects of care coordination for patients with multiple chronic conditions hold potential for higher liability. Physicians coordinating care have a broader responsibility for patients with complex conditions who have a greater chance of poor outcomes. Care coordinators may be held to a higher standard of care by adopting best practices guidelines or by making medical decisions on issues that require specialized expertise. Other aspects, however, lower liability risk: elderly patients are less likely to sue, care coordination should improve outcomes, and the information systems that support enhanced care coordination target the major sources of medical error in primary care. On balance, the liability risks of care coordination are commensurate with other risks in primary care practice. Liability insurers indicated no reluctance to insure physicians who coordinate care for patients with multiple chronic conditions and no strong tendency to attribute higher risk to this role. Physicians who currently perform these or similar functions have not encountered demonstrably higher liability. CONCLUSIONS: Physicians' perceptions about the liability risks of coordinating care for patients with multiple chronic conditions do not match evidence about the factors actually driving liability. It appears from many informed sources that there is no strong basis for physicians who perform these functions to have serious concerns about liability; instead, care coordination done well may lower liability risks. PMID- 15798037 TI - Outpatient colonoscopy by rural family physicians. AB - PURPOSE: Five percent of family physicians offer colonoscopy services, either in the office or the hospital, often in rural areas that have no gastroenterologist. Two previous large series have shown the quality and safety of colonoscopy performed by family physicians. The purpose of this study was to verify these findings in an outpatient setting, as well as to obtain patient satisfaction data. METHODS: Data were obtained from 731 colonoscopies performed between 1996 and 2001 in a rural Virginia family practice. These data included patients' age and sex, indications for the procedure, drug dosages for sedation, cecal intubation rates, pathologic findings, complications, and referral correlation findings compared with the original examination. A patient satisfaction survey was done. RESULTS: The adenoma detection rate was 27.2% for men and 21.4% for women older than age 50 years. Six adenocarcinomas and 5 large (>2 cm) villous adenomas were detected, and the patients were referred for definitive surgical resection. A total of 29 patients (4%) were referred: 10 to colorectal surgery and 19 to gastroenterology for resection of large polyps. Correlation of findings at referral with the initial examination was excellent. Cecal intubation rates increased from 89.5% from 1996-1998 to 94.6% from 1999-2001. Minor sedation complications occurred in 5 cases (<1%), and patients responded to supportive care. A high degree of satisfaction was reported by patients, with a mean satisfaction score of 8.8 on a scale from 1 to 10. CONCLUSIONS: Colonoscopy can be performed safely and competently by properly trained family physicians in an outpatient setting with a high degree of patient satisfaction. PMID- 15798040 TI - Patients' beliefs about racism, preferences for physician race, and satisfaction with care. AB - PURPOSE: Few studies have attempted to link patients' beliefs about racism in the health care system with how they use and experience health care. METHODS: Using telephone survey data from a national sample of 1,479 whites, 1,189 African Americans, and 983 Latinos, we explored patients' beliefs about racism, their preferences for the race and ethnicity of their physician, and their satisfaction with that physician. A scale was developed to reflect patients' beliefs about racism. Race-stratified analyses assessed associations between patients' beliefs, racial preferences for physicians, choice of physician, and satisfaction with care. RESULTS: Among African Americans, stronger beliefs about racial discrimination in health care were associated with preferring an African American physician (P <.001). Whereas only 22% of African Americans preferred an African American physician, those who preferred a African American physician and had an African American physician were more likely to rate their physician as excellent than did African Americans who preferred a African American physician but had a non-African American physician (57% vs 20%, P <.001). Latinos with stronger beliefs about discrimination in health care were more likely to prefer a Latino physician (P <.001). One third of Latinos preferred a Latino physician. Though not statistically significant, those who preferred and had a Latino physician rated their physician higher than Latinos who preferred a Latino physician but had a non-Latino physician (40% vs 29%). CONCLUSIONS: Many African Americans and Latinos perceive racism in the health care system, and those who do are more likely to prefer a physician of their own race or ethnicity. African Americans who have preferences are more often satisfied with their care when their own physicians match their preferences. PMID- 15798039 TI - Transferrin saturation, dietary iron intake, and risk of cancer. AB - PURPOSE: Transferrin saturation of more than 60% has been identified as a cancer risk factor. It is unclear whether dietary iron intake increases the risk of cancer among individuals with transferrin saturation of less than 60%. The purpose of this study was to examine the association of dietary iron intake and the risk of cancer among adults with increased transferrin saturation. METHODS: Analysis of a cohort study, the National Health and Nutrition Examination Survey I Epidemiologic Follow-Up Study, was performed. US adults (aged 25 to 74 years at baseline) were followed up from baseline in 1971-1974 to 1992 (N = 6,309). RESULTS: A total of 7.3% of the US population had a serum transferrin saturation of more than 45% at baseline. Intake of dietary iron was essentially uncorrelated with transferrin saturation (r = 0.04). Compared with individuals who had normal serum transferrin saturation and low dietary iron intake, individuals whose serum transferrin saturation was more than 45% and who had high dietary iron intake also had an increased adjusted relative risk of cancer (2.24; 95% confidence interval [CI], 1.02-4.89). Increased risk was not found for individuals with a transferrin saturation of more than 45% but a normal dietary iron intake (hazard ratio, 1.02; 95% CI, 0.69-1.49). Transferrin saturation levels could be set as low as 41%, and the individuals with high transferrin saturation and high dietary iron intake would still have an increased adjusted relative risk of cancer (hazard ratio, 2.00; 95% CI, 1.04-3.82). CONCLUSIONS: Among persons with increased transferrin saturation, a daily intake of dietary iron more than 18 mg is associated with an increased risk of cancer. Future research might focus on the benefits of dietary changes in those individuals with increased serum transferrin saturation. PMID- 15798041 TI - Opening the black box: cognitive strategies in family practice. AB - PURPOSE: We wanted to describe the cognitive strategies used by family physicians when structuring the decision-making tasks of an outpatient visit. METHODS: This qualitative study used cognitive task analysis, a structured interview method in which a trained interviewer works individually with expert decision makers to capture their stages and elements of information processing. RESULTS: Eighteen family physicians of varying levels of experience participated. Three dominant themes emerged: time pressure, a high degree of variation in task structuring, and varying degrees of task automatization. Based on these data and previous research from the cognitive sciences, we developed a model of novice and expert approaches to decision making in primary care. The model illustrates differences in responses to unexpected opportunity in practice, particularly the expert's use of attentional surplus (reserve capacity to handle problems) vs the novice's choice between taking more time or displacing another task. CONCLUSIONS: Family physicians have specific, highly individualized cognitive task-structuring approaches and show the decision behavior features typical of expert decision makers in other fields. This finding places constraints on and suggests useful approaches for improving practice. PMID- 15798045 TI - Screening for glaucoma: recommendation statement. PMID- 15798043 TI - Interpersonal continuity of care and care outcomes: a critical review. AB - PURPOSE: We wanted to undertake a critical review of the medical literature regarding the relationships between interpersonal continuity of care and the outcomes and cost of health care. METHODS: A search of the MEDLINE database from 1966 through April 2002 was conducted by the primary author to find original English language articles focusing on interpersonal continuity of patient care. The articles were then screened to select those articles focusing on the relationship between interpersonal continuity and the outcome or cost of care. These articles were systematically reviewed and analyzed by both authors for study method, measurement technique, and quality of evidence. RESULTS: Forty-one research articles reporting the results of 40 studies were identified that addressed the relationship between interpersonal continuity and care outcome. A total of 81 separate care outcomes were reported in these articles. Fifty-one outcomes were significantly improved and only 2 were significantly worse in association with interpersonal continuity. Twenty-two articles reported the results of 20 studies of the relationship between interpersonal continuity and cost. These studies reported significantly lower cost or utilization for 35 of 41 cost variables in association with interpersonal continuity. CONCLUSIONS: Although the available literature reflects persistent methodologic problems, it is likely that a significant association exists between interpersonal continuity and improved preventive care and reduced hospitalization. Future research in this area should address more specific and measurable outcomes and more direct costs and should seek to define and measure interpersonal continuity more explicitly. PMID- 15798044 TI - Screening for primary open-angle glaucoma in the primary care setting: an update for the US preventive services task force. AB - PURPOSE: Primary open-angle glaucoma (POAG) is a leading cause of blindness and vision-related disability. This review examines the effectiveness of screening for and treatment of early POAG in asymptomatic persons. METHODS: We identified studies of glaucoma screening and treatment from MEDLINE, the Cochrane Library, and glaucoma experts. Two reviewers abstracted relevant studies and graded articles according to US Preventive Services Task Force criteria. RESULTS: No randomized, controlled trials of population screening for POAG have been reported. Two randomized controlled trials compared the efficacy of treatment to lower intraocular pressure with no treatment for persons who have early primary open-angle glaucoma. In a Swedish trial, treatment reduced progression at 5 years from 62% without treatment to 45% with treatment (absolute risk reduction [ARR] 17%, number needed to treat 5.8, P = .007). In a US trial of patients with early POAG and normal intraocular pressure, progression at 5 years was observed in 39% of those without treatment and 33% of those with treatment (P = .21). The benefit of delaying progression of visual field loss on vision-related function in patients with early POAG is unclear. The principal harm of treatment is loss of visual acuity resulting from an increased risk of cataract formation. CONCLUSIONS: Treatment to lower intraocular pressure may delay progression of visual field deficits in some asymptomatic individuals with early POAG. Further studies of population screening are needed to show that early recognition and treatment of glaucoma in asymptomatic patients are effective in improving vision specific functional outcomes and health-related quality of life. PMID- 15798042 TI - The practice of acupuncture: who are the providers and what do they do? AB - PURPOSE: This study provides basic information about the training and practices of licensed acupuncturists. METHODS: Randomly selected licensed acupuncturists in Massachusetts and Washington state were interviewed and asked to record information on 20 consecutive patient visits. RESULTS: Most acupuncturists in both states had 3 or 4 years of academic acupuncture training and had received additional "postgraduate" training as well. Acupuncturists treated a wide range of conditions, including musculoskeletal problems (usually back, neck, and shoulder) (33% in Massachusetts and 47% in Washington), general body symptoms (12% and 9%, respectively) such as fatigue, neurological problems (10% and 12%, respectively) (eg, headaches), and psychological complaints (10% and 8%, respectively) (especially anxiety and depression). Traditional Chinese medicine (TCM) was the predominant style of acupuncture used in both states (79% and 86%, respectively). Most visits included a traditional diagnostic assessment (more than 99%), regular body acupuncture (95% and 93%, respectively), and additional treatment modalities (79% and 77%, respectively). These included heat and lifestyle advice (66% and 65%, respectively), most commonly dietary advice and exercise recommendations. Chinese herbs were used in about one third of visits. Although most patients self-referred to acupuncture, about one half received concomitant care from a physician. Acupuncturists rarely communicated with the physicians of their patients who were providing care for the same problem. CONCLUSIONS: This study contributes new information about acupuncturists and the care they provide that should be useful to clinicians interested in becoming more knowledgeable about complementary or alternative medical therapies available to their patients. PMID- 15798046 TI - Ecological change and the future of the human species: can physicians make a difference? AB - Global environmental change is occurring so rapidly that it is affecting the health and threatening the future of many of Earth's inhabitants, including human beings. Global warming; contamination of the air, water, and soil; and rampant deforestation have led to a collapse in biodiversity that threatens the integrity of the biophysical systems upon which all organisms depend.A basic cause of environmental degradation is human overpopulation and the nonsustainable consumption of natural resources by the human community. Everything that we have accomplished in the fields of medicine and public health could be undermined if we do not pay attention to these rapid environmental changes. As healers, human beings, and members of the biological community, we need to broaden our perspective on health and disease. Unless we devote our attention to stabilizing and repairing the ecosystem, our professional and personal accomplishments as health professionals may be swept away. Health care providers--particularly physicians--can play a role by adopting an ecosystem health perspective as we ply our trade. By helping people avoid unwanted pregnancies, by using resources parsimoniously, and by staying engaged in the natural world, we can help to prevent the collapse of the biological systems upon which we all depend. PMID- 15798047 TI - Academic family medicine chairs. PMID- 15798049 TI - Patient's Voice in Washington. PMID- 15798048 TI - 2005 Examination dates and test centers, 2004 Pisacano Scholars. PMID- 15798050 TI - Imaging evaluation of pulmonary and abdominal complications following hematopoietic stem cell transplantation. AB - Hematopoietic stem cell transplantation is used to treat hematologic disorders and as an adjunct treatment for solid organ malignancies. After undergoing transplantation, patients are at risk for opportunistic infections and other complications caused by dysfunction of the immune system. Pulmonary complications include cryptogenic organizing pneumonia, opportunistic pneumonias caused by Aspergillus and Zygomycetes species and cytomegalovirus, alveolar hemorrhage, and constrictive bronchiolitis. Abdominal complications include hepatic veno occlusive disease, graft-versus-host disease (GVHD), colitis, and hemorrhagic cystitis. Allogeneic transplant recipients are at risk for developing GVHD. Autologous and syngeneic transplant recipients are less likely to have chronic or late posttransplantation complications. Nonmyeloablative transplant recipients are less likely to develop opportunistic infections and other complications in the period immediately following transplantation, but are at risk for developing chronic GVHD and other chronic complications. Radiologic evaluation serves as the cornerstone for timely diagnosis of these complications, which is essential to reduce patient morbidity and mortality. Combining clinical factors-including the type of transplant and the point of time during the posttransplantation course with characteristic imaging features yields the most specific and accurate differential diagnosis for radiologic findings in stem cell transplant recipients. PMID- 15798052 TI - Fat-containing lesions of the liver: radiologic-pathologic correlation. AB - Fat-containing tumors of the liver are a heterogeneous group of tumors with characteristic histologic features, variable biologic profiles, and variable imaging findings. Benign liver lesions that contain fat include focal or geographic fatty change (steatosis), pseudolesions due to postoperative packing material (omentum), adenoma, focal nodular hyperplasia, lipoma, angiomyolipoma, cystic teratoma, hepatic adrenal rest tumor, pseudolipoma of the Glisson capsule, and xanthomatous lesions in Langerhans cell histiocytosis. Malignant liver lesions that can contain fat include hepatocellular carcinoma, primary and metastatic liposarcoma, and hepatic metastases. Identification of fat within a liver lesion can be critical in characterization of the lesion. The imaging characteristics of a lesion coupled with the pattern of intratumoral fatty change are helpful in narrowing the differential diagnosis. Although the presence of fat can be demonstrated with computed tomography or ultrasound, magnetic resonance imaging is the most specific imaging technique for demonstration of both microscopic and macroscopic fat. PMID- 15798053 TI - Real-time harmonic contrast material-specific US of focal liver lesions. AB - Recent advances in contrast material-enhanced ultrasonography (US) mainly include (a) development of low-acoustic-pressure (low-mechanical-index) harmonic software, capable of obtaining real-time images without disrupting contrast material microbubbles, and (b) commercialization of new contrast media ("second generation" contrast media), capable of producing intense echo signals in this low-mechanical-index setting. With use of low-mechanical-index continuous-mode contrast-enhanced US, the circulatory kinetic models of various focal liver lesions can be displayed dynamically. Hepatic lesions usually have typical perfusion characteristics and enhancement patterns through the various phases of parenchymal enhancement, which helps characterize lesions and, in most cases, allows definitive diagnosis, even among lesions that exhibit very similar baseline appearances. Because of the use of harmonic technologies at low emission frequencies, there is some loss of spatial resolution and overall image quality, typically resulting in a grainy appearance. In addition, lesion depth affects the detectability of vascularity to some degree in that poor signal arises from deep seated lesions. Moreover, liver attenuation (eg, in patients with steatosis or chronic liver disease) further reduces the sensitivity of contrast-enhanced US. Nevertheless, with its unique capacity to provide images in real time, low mechanical-index contrast-enhanced US is the dynamic imaging modality of choice in the differential diagnosis of focal liver lesions. PMID- 15798054 TI - Mesenteric lymph nodes seen at imaging: causes and significance. AB - With the advent of multidetector computed tomography, routine evaluation of mesenteric lymph nodes is now possible. For the first time, normal mesenteric nodes may be reliably identified noninvasively. Because of the increasing volume of cross-sectional imaging examinations being performed, lymph nodes in the mesentery are being detected with increasing frequency. This is often an unsuspected finding. Although the detected lymph nodes may be normal, there is a large number of disease processes that may lead to mesenteric lymphadenopathy. The most common causes of mesenteric lymphadenopathy are neoplastic, inflammatory, and infectious processes. Many of these causes may also result in lymphadenopathy elsewhere in the body. It is important to recognize mesenteric lymphadenopathy in patients with a history of a primary carcinoma because the lymphadenopathy affects the staging of the disease, which in turn will affect further management. In addition, mesenteric lymphadenopathy may be the only indicator of an underlying inflammatory or infectious process causing abdominal pain. The distribution of the lymph nodes may indicate the exact nature of the underlying disease process, and the correct treatment may then be instituted. Besides neoplastic, inflammatory, and infectious processes, many other disease processes may occasionally result in mesenteric lymphadenopathy. PMID- 15798055 TI - Of fractures, hip and hangman, of gamekeepers and red herrings. PMID- 15798056 TI - Neoplasms of the renal medulla: radiologic-pathologic correlation. AB - Tumors of the renal medulla cover a wide spectrum, with characteristic histomorphology and variable biologic profiles. Renal medullary tumors can be categorized into benign and malignant neoplasms based on histologic features and clinico-biologic behavior. They can be further classified into pediatric and adult tumors based on the patient age group. When small, renal medullary tumors may be differentiated from the more common renal adenocarcinomas by their central location and certain demographic characteristics. Although most large malignant medullary tumors demonstrate imaging findings that are indistinguishable from those of other renal malignancies, some tumors demonstrate imaging findings that may suggest a specific diagnosis. PMID- 15798057 TI - Rheumatoid arthritis: a practical guide to state-of-the-art imaging, image interpretation, and clinical implications. AB - Rheumatoid arthritis (RA) is a chronic systemic disease of unknown origin that predominantly involves synovial tissue. RA affects 0.5%-1.0% of the global population, with females affected more frequently than males. Early diagnosis and initiation of proper therapy help modify the course of the disease and reduce the degree of severe late sequelae. Radiology plays a key role in diagnosis and management of RA. Currently, magnetic resonance imaging is the best imaging modality because it depicts soft-tissue changes and damage to cartilage and bone even better and at an earlier stage than does computed tomography. Ultrasound and conventional radiography are more readily available but cannot show the entire spectrum of the disease. Diagnosis and differential diagnosis are achieved by identifying certain radiologic parameters, which are also used for grading purposes. The disease does not follow a linear course, especially with the early initiation of potent therapy. Knowledge of the imaging findings enables the radiologist to accurately select the most helpful imaging technique. Familiarity with the pathophysiologic mechanisms of RA, the imaging findings, and the grading systems and a basic knowledge of therapeutic regimens are prerequisites for a tailored diagnostic approach by the radiologist. PMID- 15798058 TI - Avascular necrosis of the talus: a pictorial essay. AB - The talus is predisposed to avascular necrosis (AVN), or bone death due to ischemia, owing to its unique structure, characteristic extraosseous arterial sources, and variable intraosseous blood supply. Both traumatic and atraumatic causes have been implicated in talar AVN. The risk of posttraumatic AVN can be predicted using the Hawkins classification system. In addition, the "Hawkins sign" can be used as a radiographic marker that excludes the development of AVN. At radiography, talar AVN typically manifests as an increase in talar dome opacity (sclerosis), followed by deformity and, in severe cases, articular collapse and bone fragmentation. At any stage of this sequence, the radiographic findings can vary depending on differences in the vascular status of the talus and the degree of bone repair. Magnetic resonance imaging is the most sensitive technique for detecting talar AVN and can be used when AVN is strongly suspected clinically despite normal radiographic findings. Computed tomography (CT) also demonstrates typical patterns and can be used to confirm radiographic findings. Coronal CT is required for viewing the articular surface of the talar dome to rule out subtle depression, collapse, and fragmentation. Nevertheless, radiography remains the mainstay of the diagnosis and temporal observation of talar AVN. PMID- 15798059 TI - Radiologic evaluation of uncommon inflammatory and reactive breast disorders. AB - The radiologic features of breast lesions caused by immunologic, reactive, and noncurrent infectious diseases often mimic those of malignancy, frequently constituting a diagnostic challenge even if the underlying disease is known. Churg-Strauss syndrome mimics carcinomatous mastitis. Amyloidosis usually manifests as a suspicious mass, often accompanied by microcalcifications. Wegener granulomatosis and sarcoidosis often manifest as irregular masses, although sarcoidosis can also manifest as round, well-defined masses reflecting intramammary node involvement. Diabetic mastopathy is a rare but well-known entity in patients with long-standing insulin-dependent diabetes. Breast involvement by necrobiotic xanthogranulomatosis is rare and manifests as multiple bilateral asymmetric lesions. Multiple clustered hypoechoic tubular structures in a large hypoechoic mass seen after pregnancy can be suggestive of granulomatous mastitis. Mammary tuberculosis can manifest with a nodular, diffuse, or sclerosing pattern. A granulomatous inflammatory reaction must be carefully evaluated because it constitutes the major feature of a diverse group of diseases that includes vasculitis, granulomatous mastitis, tuberculosis, and carcinoma associated sarcoidlike reactions. Core biopsy can play a major role in developing a differential diagnosis for these rare immunologic, inflammatory, or infectious disorders affecting the breast, and knowledge of these entities can, in the appropriate clinical setting, help the radiologist narrow the differential diagnosis, although cancer must be excluded definitively. PMID- 15798060 TI - Pitfalls in 16-detector row CT of the coronary arteries. AB - Recently developed 16-detector row computed tomography (CT) has been introduced as a reliable noninvasive imaging modality for evaluating the coronary arteries. In most cases, with appropriate premedication that includes beta-blockers and nitroglycerin, ideal data sets can be acquired from which to obtain excellent quality coronary CT angiograms, most often with multiplanar reformation, thin slab maximum intensity projection, and volume rendering. However, various artifacts associated with data creation and reformation, postprocessing methods, and image interpretation can hamper accurate diagnosis. These artifacts can be related to pulsation (nonassessable segments, pseudostenosis) as well as rhythm disorders, respiratory issues, partial volume averaging effect, high-attenuation entities, inappropriate scan pitch, contrast material enhancement, and patient body habitus. Some artifacts have already been resolved with technical advances, whereas others represent partially inherent limitations of coronary CT angiography. Familiarity with the pitfalls of coronary angiography with 16 detector row CT, coupled with the knowledge of both the normal anatomy and anatomic variants of the coronary arteries, can almost always help radiologists avoid interpretive errors in the diagnosis of coronary artery stenosis. PMID- 15798062 TI - Anatomic pitfalls of the heart and pericardium. AB - Recent technologic advances have led to more frequent dedicated cross-sectional imaging of the heart. Faster scanning techniques, cardiac gating, and advanced postprocessing software allow improved visualization of finer anatomic details of the heart and pericardium compared with older techniques and software. Use of thin-section computed tomography (CT) or image reformatting in nonaxial planes may be helpful in some cases. The cardiac and pericardial structures are usually readily demonstrated with CT, even if chest CT is performed for evaluation of noncardiac structures. However, radiologists are expected to evaluate all structures on an image, and incidental findings are common. Radiologists must first be familiar with the normal anatomic structures of the heart and pericardium (eg, atria, ventricles, cardiac valves, pericardial recesses, paracardiac structures) to avoid mistaking them for pathologic processes. PMID- 15798063 TI - From the archives of the AFIP: abdominal neoplasms in patients with neurofibromatosis type 1: radiologic-pathologic correlation. AB - Neurofibromatosis type 1 (NF1) is one of the most common genetic disorders. NF1 is a complex disease resulting from a spectrum of mutations that may occur at many locations along the large, complex NF1 gene, which is located on chromosome 17. Mutations of the NF1 gene lead to abnormal tumor suppression. Consequently, patients with NF1 have an increased prevalence of benign and malignant neoplasms throughout the body. There are five categories of NF1 tumors that occur in the abdomen: neurogenic, neuroendocrine, nonneurogenic gastrointestinal mesenchymal, embryonal, and miscellaneous. Many of these tumors are age related, occur at specific anatomic locations, and have unique imaging features. Notably, many patients have a variety of organs affected because there is a high prevalence of multiple tumors occurring in the same patient. Neurofibromas are the most common benign neoplasms and may occur in the retroperitoneum or visceral organs. Malignant peripheral nerve sheath tumor is an aggressive malignancy that is the most common malignant tumor of the abdomen in patients with NF1. Interpreting abdominal imaging studies in patients with NF1 can be challenging because of the wide spectrum and diverse nature of tumors that occur in this disease. PMID- 15798064 TI - Best cases from the AFIP: intrarenal teratoma. PMID- 15798065 TI - Best cases from the AFIP: supratentorial ependymoma. PMID- 15798066 TI - AAPM/RSNA physics tutorial for residents: technological and psychophysical considerations for digital mammographic displays. AB - Digital mammography is gradually replacing screen-film analog mammography, a transition driven by a desire to improve the efficiency and possibly the quality of the interpretation of mammograms. Digital mammography involves the use of electronic display devices to display the mammograms. Currently, two electronic technologies are used to display digital mammograms: the cathode-ray tube (CRT) and liquid crystal display (LCD). CRT and LCD devices have imaging characteristics markedly different from those of transilluminated film, which has conventionally been used to display analog mammograms. Consequently, the transition to digital mammography necessitates consideration of a number of psychophysical factors pertaining to effective display of mammograms. Some of these factors are related to specific performance characteristics of the display devices, whereas others are related to inherent characteristics of the human visual system. The main psychophysical factors that affect the interpretation of medical images are contrast, resolution, and noise. Optimal display of mammograms is achieved by taking these factors into consideration and by using time efficient, intuitive, and reader-specific user interfaces. Because display devices are susceptible to variations in hardware and calibration and to degradation over time, acceptance testing and quality control testing are necessary to maintain an adequate level of display quality. PMID- 15798067 TI - Medical devices of the abdomen and pelvis. AB - Medical devices in the abdomen and pelvis are probably less frequently seen than those in the chest or extremities, but they are important and should be recognized. These devices can be grouped into a few major categories: intestinal tubes, genitourinary devices, postoperative apparatus, and a wide variety of odds and ends. Many of these devices are used to monitor or treat gastrointestinal and genitourinary disease. Some of them, such as inferior vena cava filters and drug infusion pumps, treat systemic problems, and some of them are devices used in treating another anatomic region. It is strongly recommended that scout views for chest, abdominal, and pelvic computed tomographic studies be carefully examined for medical apparatus. Medical devices are often more easily recognized on scout images, and their inappropriate locations and complications can be better appreciated on the subsequent cross-sectional images if one is alerted to their presence in the first place. The evaluation of routine medical devices should be considered as important as any other aspect of a radiologic examination. PMID- 15798068 TI - Informatics in radiology (infoRAD): new tools for computer assistance in thoracic CT. Part 1. Functional analysis of lungs, lung lobes, and bronchopulmonary segments. AB - Owing to the rapid development of scanner technology, thoracic computed tomography (CT) offers new possibilities but also faces enormous challenges with respect to the quality of computer-assisted diagnosis and therapy planning. In the framework of the Virtual Institute for Computer Assistance in Clinical Radiology cooperative research project, a prototypical software application was developed to assist the radiologist in functional analysis of thoracic CT data. By identifying the anatomic compartments of the lungs, the software application enables assessment of established functional CT parameters for each individual lung, pulmonary lobe, and pulmonary segment. Such region-based assessment allows a more localized diagnosis of lung diseases such as emphysema and more accurate estimation of regional lung function from CT data. With close cooperation between computer scientists and radiologists, the software application was tested and optimized to achieve a high degree of usability. Several clinical studies were carried out, the results of which indicated that the software application improves quantification in diagnosis, therapy planning, and therapy monitoring with respect to accuracy and time required. PMID- 15798069 TI - Informatics in radiology (infoRAD): survey of personal digital assistant use in radiology. AB - There has been widespread adoption of personal digital assistants (PDAs) within medicine in recent years. However, information on the prevalence and usage of these devices among radiologists is limited. A survey was designed and mailed to randomly selected members of the Radiological Society of North America to determine the percentage of PDA users, their use patterns, and the types of applications that they would like to see in the future. The use patterns of attending radiologists were compared with those of trainees (residents and fellows). Overall usage was also compared with the relevant findings in two surveys of internal medicine users. It was found that slightly less than one-half of respondents used PDAs on a daily basis, a finding that was comparable to that in the internal medicine surveys. However, less than one-quarter of PDA users had radiology-specific applications installed on their devices, whereas a greater percentage of internal medicine users had software such as drug databases and clinical references on their PDAs. Radiology trainees had a higher rate of both PDA ownership and radiology application usage than did attending radiologists. It is likely that, as PDA hardware becomes more powerful, with higher display resolution, better wireless networking capabilities, and greater memory capacity, PDA ownership as well as radiology application usage will increase. PMID- 15798070 TI - Informatics in radiology (infoRAD): integrating MIRC-compliant semiautomated teaching files into PACS work flow. AB - A Medical Image Resource Center (MIRC)-compliant teaching file system was created that can be integrated into a picture archiving and communication system (PACS) environment. This system models the three-step process necessary for efficient teaching file creation in a PACS environment: (a) identifying and transferring a case quickly and easily during primary interpretation, (b) editing and authoring the case outside of primary interpretation time, and (c) publishing the case locally and via MIRC standard-based mechanisms. Images from interesting cases are e-mailed to the teaching file system from either the PACS workstation or the radiologist's personal computer. Notes and clinical information may be included in the e-mail text to prompt the recollection of case details. Images are automatically extracted from the e-mail and sent to an image repository, and text fields are captured in a database. The World Wide Web-based authoring component provides tools for final authoring of cases and for the manipulation of existing cases. Authors designate access levels for each case, which is then made available locally and, potentially, to the entire MIRC-compliant community. Although this application has not yet been implemented as a departmental solution, it promises to improve and streamline medical education and promote better patient care. PMID- 15798071 TI - Scenes from the past: sun-dried X rays. PMID- 15798074 TI - A review of family and social determinants of children's eating patterns and diet quality. AB - With the growing problem of childhood obesity, recent research has begun to focus on family and social influences on children's eating patterns. Research has demonstrated that children's eating patterns are strongly influenced by characteristics of both the physical and social environment. With regard to the physical environment, children are more likely to eat foods that are available and easily accessible, and they tend to eat greater quantities when larger portions are provided. Additionally, characteristics of the social environment, including various socioeconomic and sociocultural factors such as parents' education, time constraints, and ethnicity influence the types of foods children eat. Mealtime structure is also an important factor related to children's eating patterns. Mealtime structure includes social and physical characteristics of mealtimes including whether families eat together, TV-viewing during meals, and the source of foods (e.g., restaurants, schools). Parents also play a direct role in children's eating patterns through their behaviors, attitudes, and feeding styles. Interventions aimed at improving children's nutrition need to address the variety of social and physical factors that influence children's eating patterns. PMID- 15798075 TI - Beverage consumption patterns in elementary school aged children across a two year period. AB - OBJECTIVE: Existing data was reexamined to determine changes in beverage consumption and associations between beverages consumed and BMI Z-score in children (n = 164) across two years. METHODS: Beverages (milk, 100% juice, diet soda or sugar sweetened) and total caloric intake were calculated from a 24-hour diet recall. Height and weight were measured to calculate BMI. Subjects were categorized by BMI Z-score as normal weight, overweight, gained weight and lost weight. Data was collected at baseline and year 2. RESULTS: Significant decreases in milk and increases in diet soda were found over two years in all subjects and normal weight, whereas overweight had a significant increase in diet soda consumption and a decrease in milk consumption that did not reach significance. Change in milk consumption was inversely correlated with sugar-sweetened beverage consumption. Increases in diet soda consumption were significantly greater for overweight and subjects who gained weight as compared to normal weight subjects. Baseline BMI Z-score and year 2 diet soda consumption predicted 83.1% of the variance in year 2 BMI Z-score. CONCLUSION: Shifts in beverage consumption were found in this convenient sample across two years. Diet soda consumption was the only type of beverage associated with year 2 BMI Z-score, and consumption was greater in overweight subjects and subjects who gained weight as compared to normal weight subjects at two years. Additional longitudinal data examining associations between beverage consumption and BMI is needed in children and adolescents, as consumption of regular and diet soda has become more of a social norm. PMID- 15798077 TI - Effect of iron-fortified drinking water of daycare facilities on the hemoglobin status of young children. AB - BACKGROUND: Anemia is the most prevalent nutrition problem in young children. One possible strategy to prevent anemia is affordable fortification of drinking water. OBJECTIVE: The aim of this study was to evaluate the impact of iron fortified drinking water of daycare facilities on the hemoglobin and anthropometric status of pre-school children. DESIGN: Hemoglobin (Hb) status, weight and height measurements were assessed in 160 pre-school children aged 6 to 59 m before and after 8 m consumption of iron- (12 mg/L) and vitamin C- (90 mg/L) fortified drinking water. RESULTS: Initially, 43.2% (69) of the children evaluated as being anemic decreased to 21% (37) at the end of study. At baseline, 42 (26.3%) children suffered from moderate anemia and 27 (16.9%) suffered severe anemia, but after iron fortification, total number of children suffering from moderate and severe anemia had decreased to 32 (20.7%) and 5 (3%), respectively. Weight-for-age (WAZ), height-for-age (HAZ) and weight-for-height (WHZ) Z-scores increased significantly from -0.84 +/- 1.03 to 0.06 +/- 1.10, -0.84 +/- 1.11 to 0.54 +/- 1.10 and -0.39 +/- 0.94 to -0.18 +/- 1.14, respectively (p < 0.05). Daycare personnel reported increased appetite and food consumption and decreased absenteeism during intervention. CONCLUSION: Daily consumption of iron-fortified drinking water in daycare facilities is an effective, simple and inexpensive means of reducing and controlling for moderate and severe anemia in pre-school children. PMID- 15798076 TI - Potassium intake and the calcium economy. AB - BACKGROUND: Dietary potassium intake (K) lowers urinary calcium (Ca) excretion and, in short-term studies, may improve Ca balance. PURPOSE: Our objective was to assess K effects on the Ca economy under steady-state conditions. DESIGN: 8-day, inpatient metabolic studies of nitrogen, phosphorus, and Ca balance, combined with dual isotopic Ca tracer kinetics studies. Study diet matched to prestudy nutrient intakes. SUBJECTS: 191 single women studied from 1-5 times at approximately 5-year intervals, for a total of 644 inpatient studies. Median age at time of study: 50.2 yrs; 301 studies were performed postmenopausally without hormone replacement; 343 were either premenopausal or postmenopausal but on estrogen replacement therapy. RESULTS: Dietary K was highly significantly associated with urinary Ca excretion, with a coefficient of -0.0109 mmol urine Ca/mmol diet K. However, dietary K was negatively correlated with dual-tracer Ca absorption (coefficient for Ca absorption fraction: -0.00094/mmol dietary K), and was not associated with urine Ca after adjustment for Ca absorption. CONCLUSION: While a high K diet (i.e., one rich in fruits, vegetables, and dairy products) has multiple health benefits and clearly lowers urine Ca, it does not seem to exert any appreciable net influence on the Ca economy, largely because the reduced calciuria is offset by reduction of intestinal absorption. We note, however, that since the high K intakes in our studies come more from milk and meat than from fruits and vegetables, we cannot exclude a possible balance effect for different food sources of K. PMID- 15798078 TI - Dietary patterns of Korean Americans described by factor analysis. AB - OBJECTIVES: This study tested the hypothesis that the dietary behaviors of Korean Americans (KAs) can be grouped into dietary patterns and subsequently examined relations between major dietary patterns and sociodemographic characteristics. METHODS: A cross-sectional study was conducted with KAs residing in Michigan using a mail survey. Of 1,860 questionnaires mailed to KAs in Michigan, 637 (34%) responded and 497 first-generation KAs (263 men, 234 women, aged 30-87 y) were included in the final analyses. A factor analysis was used to identify dietary patterns from a 93-item food frequency questionnaire developed for KAs (KFFQ). RESULTS: Major dietary patterns of KAs were labeled "vegetable/fruit," and "traditional Korean" in both men and women and "acculturated American" in men and "traditional American" in women. The "traditional Korean" dietary pattern was negatively associated with length of residence in the U.S. for both men and women (p < 0.01). The other major dietary patterns were not associated with other sociodemographic variables examined in men; however, the "vegetable/fruit" dietary pattern was positively associated with length of residence in the U.S. (p < 0.05) and education level (p < 0.05) in women. CONCLUSIONS: Dietary pattern analysis can be used to understand dietary behaviors regarding health risks of ethnically different immigrants including KAs. PMID- 15798080 TI - Dietary protein to support anabolism with resistance exercise in young men. AB - Resistance exercise is fundamentally anabolic and as such stimulates the process of skeletal muscle protein synthesis (MPS) in an absolute sense and relative to skeletal muscle protein breakdown (MPB). However, the net effect of resistance exercise is to shift net protein balance (NPB = MPS - MPB) to a more positive value; however, in the absence of feeding NPB remains negative. Feeding stimulates MPS to an extent where NPB becomes positive, for a transient time. When combined, resistance exercise and feeding synergistically interact to result in NPB being greater than with feeding alone. This feeding- and exercise-induced stimulation of NPB is what, albeit slowly, results in muscle hypertrophy. With this rudimentary knowledge we are now at the point where we can manipulate variables within the system to see what impact these interventions have on the processes of MPS, MPB, and NPB and ultimately and perhaps most importantly, muscle hypertrophy and strength. We used established models of skeletal muscle amino acid turnover to examine how protein source (milk versus soy) acutely affects the processes of MPS and MPB after resistance exercise. Our findings revealed that even when balanced quantities of total protein and energy are consumed that milk proteins are more effective in stimulating amino acid uptake and net protein deposition in skeletal muscle after resistance exercise than are hydrolyzed soy proteins. Importantly, the finding of increased amino acid uptake would be independent of the differences in amino acid composition of the two proteins. We propose that the improved net protein deposition with milk protein consumption is also not due to differences in amino acid composition, but is due to a different pattern of amino acid delivery associated with milk versus hydrolyzed soy proteins. If our acute findings are accurate then we hypothesized that chronically the greater net protein deposition associated with milk protein consumption post-resistance exercise would eventually lead to greater net protein accretion (i.e., muscle fiber hypertrophy), over a longer time period. In young men completing 12 weeks of resistance training (5d/wk) we observed a tendency (P = 0.11) for greater gains in whole body lean mass and whole as greater muscle fiber hypertrophy with consumption of milk. While strength gains were not different between the soy and milk-supplemented groups we would argue that the true significance of a greater increase in lean mass that we observed with milk consumption may be more important in groups of persons with lower initial lean mass and strength such as the elderly. PMID- 15798079 TI - Costa Rican adolescents have a deleterious nutritional profile as compared to adults in terms of lower dietary and plasma concentrations of antioxidant micronutrients. AB - OBJECTIVE: Most prospective studies in adult populations show an inverse association between consumption of fruits and vegetables and the risk of cardiovascular disease and all-cause mortality. Fruits and vegetables are rich in antioxidants. The objective was to determine whether nutrient intakes and plasma concentrations of antioxidants of adolescents are similar to those of their parents or grandparents as a way of assessing whether dietary habits of parents are passed on to their adolescent children. METHODS: Parents or grandparents (n = 159) aged 29-77 y were selected from the participants of the myocardial case control study in Costa Rica. One adolescent child or grandchild (12-20 y) was identified for each parent or grandparent. Nutrient intakes and plasma antioxidant concentrations were assessed using a validated food-frequency questionnaire and by high performance liquid chromatography, respectively. RESULTS: As compared to adults, adolescents consumed greater quantities of fruit, dairy products, red meat, carbonated drinks and polyunsaturated fat, but lower quantities of fiber and micronutrients (carotenoids, vitamins A and B6, folate, potassium, magnesium and zinc). Plasma concentrations of all carotenoids, except for lycopene, were lower in adolescents than in adults (p < 0.05). Intakes of lycopene (r = 0.16, p = 0.05), alpha-tocopherol (r = 0.19, p = 0.03) and gamma tocopherol (r = 0.21, p = 0.01) in adolescents were correlated with those of adults. Plasma concentrations of lycopene (r = 0.20, p = 0.02), alpha-tocopherol (r = 0.18, p = 0.03) and gamma-tocopherol (r = 0.28, p = 0.001) in adolescents were also correlated with those of adults. These data may suggest that adolescents may acquire habitual consumption of foods rich in micronutrients from adults. CONCLUSION: As a proportion of energy, diets of Costa Rican adolescents contain less micronutrients than those of their adult relatives. Parents should be aware of this trend and its association with poor health status. PMID- 15798082 TI - Soy protein isolate and protection against cancer. AB - OBJECTIVE: Results from epidemiological and animal studies suggest that consuming soy-containing diets reduces the incidence of certain cancers. The purpose of this presentation was to evaluate the potential of soy protein to prevent occurrence of prostate, breast and colon cancer. METHODS: Meta-analyses of published epidemiologic studies associating cancer risk with soy intake were performed. The incidence of chemically-induced mammary or colon tumors was determined for rats fed AIN-93G diets made with either casein or soy protein isolate (SPI). Western and Northern blot and microarray analyses were performed on rat mammary and colon tissues to study mechanisms underlying the effects of soy. RESULTS: Meta-analyses revealed reductions in the mean overall risk estimate for mammary (0.78, p < 0.001), colon (0.70, p < 0.001) and prostate (0.66, p < 0.001) cancer for soy consumers. The incidence of AOM-induced colon tumors and DMBA-induced mammary tumors was reduced (p < 0.05) in rats fed SPI-containing diets. Lower incidence of mammary tumors in SPI-fed rats was associated with: 1) reduced terminal end bud numbers (p < 0.05), 2) lower expression of the phase I enzyme CYP1B1 (p < 0.05) and 3) reduced expression of the Ah Receptor and ARNT (p < 0.05). CONCLUSIONS: SPI may protect against cancer via multiple mechanisms, including: 1) increased mammary gland differentiation, 2) decreased activation of procarcinogens to carcinogens and 3) regulation of genes in signal transduction pathways underlying tumor initiation, promotion and/or progression. PMID- 15798081 TI - Role of protein and amino acids in the pathophysiology and treatment of sarcopenia. AB - Sarcopenia, the loss of skeletal muscle mass and function with aging, is a multifactorial condition that slowly develops over decades and becomes a significant contributor to disability in the older population. Malnutrition and alterations in the muscle anabolic response to nutritional stimuli have been identified as potentially preventable factors that may significantly contribute to sarcopenia. Thus, nutritional interventions may be useful for the prevention and treatment of sarcopenia. PMID- 15798083 TI - Optimal quantity and composition of protein for growing children. AB - Children have distinct nutritional needs relative to growth. Adequate intakes of energy and essential amino acids are necessary for optimal deposition of lean body mass and normal growth in young children. However, there are limited data concerning protein needs of children. Most recommendations for children represent an interpolation of data derived from infants and adults. Indeed, current protein requirements for young children, while scientifically based, are estimates at best. Historically, protein status in children was evaluated using classic nitrogen balance protocols. This work indicates that a wide range of protein intakes (0.6-2.9 g/kg) can be considered adequate for young, growing children. The ability of nitrogen balance studies to accurately reflect protein utilization has been examined and it appears that further investigations of protein utilization in children using stable isotope methodology, as well as traditional nitrogen balance protocols, are necessary to better evaluate protein needs of growing children. In addition, protein source may be an important factor in optimal diet design for growing children. PMID- 15798084 TI - 2-Cyano-3,12-dioxoolean-1,9-dien-28-oic acid and related compounds inhibit growth of colon cancer cells through peroxisome proliferator-activated receptor gamma dependent and -independent pathways. AB - 2-Cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO) and the corresponding methyl (CDDO-Me) and imidazole (CDDO-Im) esters induce peroxisome proliferator-activated receptor gamma (PPARgamma)-dependent transactivation in SW-480 colon cancer cells, and these responses were inhibited by small inhibitory RNA for PPARgamma. Moreover, in a mammalian two-hybrid assay using the PPARgamma(2)-VP16 fusion plasmid and GAL4-coactivator/corepressor chimeras and a construct (pGAL4) containing five tandem GAL4 response elements, CDDO, CDDO-Me, and CDDO-IM induce transactivation and PPARgamma interaction with multiple coactivators. A major difference among the three PPARgamma agonists was the higher activity of CDDO-Im to induce PPARgamma interactions with the corepressor SMRT. CDDO, CDDO-Me, and CDDO-Im inhibited SW-480, HCT-116, and HT-29 colon cancer cell proliferation at low concentrations and induced cell death at higher concentrations. Growth inhibition at lower concentrations correlated with induction of the tumor suppressor gene caveolin-1 which is known to inhibit colon cancer cell growth. Induction of caveolin-1 by CDDO, CDDO-Me, and CDDO-Im was inhibited by the PPARgamma antagonist N-(4'-aminopyridyl-2-chloro-5-nitrobenzamide (T007), whereas higher doses induced apoptosis [poly(ADP-ribose) polymerase cleavage], which was not inhibited by T007. These results illustrate that CDDO-, CDDO-Me, and CDDO-Im induce both PPARgamma-dependent and -independent responses in colon cancer cells, and activation of these pathways are separable and concentration-dependent for all three compounds. PMID- 15798085 TI - Epidermal growth factor activates nuclear factor-kappaB in human proximal tubule cells. AB - The promotion of cell survival and regeneration in acute renal failure (ARF) is important for the restitution of renal function. Epidermal growth factor (EGF) has been implicated in the regulation of cell proliferation. We provide evidence for a direct link between EGF, nuclear factor-kappaB (NF-kappaB), and cell cycle regulation (cyclin D1). EGF was found to stimulate NF-kappaB-dependent gene transcription and DNA binding. In addition, EGF stimulated cyclin D1 promoter activity as well as cyclin D1 expression. Moreover, inhibition of NF-kappaB caused a pronounced reduction of EGF-induced cyclin D1 promoter activity. Furthermore, both EGF-mediated NF-kappaB activation and cyclin D1 expression were inhibited by coexpression of super IkappaB. Taken together, these data identify NF-kappaB and cyclin D1 as downstream targets of EGF and establish a molecular link between stimulation of EGF via activation of NF-kappaB and cyclin D1 expression in human proximal tubular cells. PMID- 15798086 TI - WT1-interacting protein and ZO-1 translocate into podocyte nuclei after puromycin aminonucleoside treatment. AB - Podocyte differentiation is required for normal glomerular filtration barrier function and is regulated by the transcription factor WT1. We identified WT1 interacting protein (WTIP) and hypothesized that it functions as both a scaffold for slit diaphragm proteins and a corepressor of WT1 transcriptional activity by shuttling from cell-cell junctions to the nucleus after injury. Endogenous WTIP colocalizes with zonula occludens-1 (ZO-1) in cultured mouse podocyte adherens junctions. To model podocyte injury in vitro, we incubated differentiated podocytes with puromycin aminonucleoside (PAN; 100 microg/ml) for 24 h, which disassembled cell-cell contacts, rearranged actin cytoskeleton, and caused process retraction. Podocyte synaptopodin expression diminished after PAN treatment, consistent with podocyte dedifferentiation in some human glomerular diseases. To assess podocyte function, we measured albumin flux across differentiated podocytes cultured on collagen-coated Transwell filters. Albumin transit across PAN-treated cells increased to levels observed with undifferentiated podocytes. Consistent with our hypothesis, WTIP, as well as ZO 1, translocated from podocyte adherens junctions to nuclei in PAN-treated cells. Because WTIP is a transcriptional corepressor for WT1, we examined the effect of PAN on expression of retinoblastoma binding protein Rbbp7 (also known as RbAp46), a WT1 target gene expressed in S-shaped bodies during nephrogenesis. Rbbp7 expression in PAN-treated podocytes was reduced compared with untreated cells. In conclusion, WTIP translocates from cell-cell junctions to the nucleus in PAN treated podocytes. We suggest that WTIP monitors slit diaphragm protein assembly and shuttles into the nucleus after podocyte injury, translating changes in slit diaphragm structure into altered gene expression and a less differentiated phenotype. PMID- 15798087 TI - Regulated expression of renal and intestinal UT-B urea transporter in response to varying urea load. AB - Production, recycling, and elimination of urea are important to maintain nitrogen balance. Adaptation to varying loads of urea due to different protein intake or in renal failure may involve changes in urea transport and may possibly affect urea transporters. In this study, we examined the expression of the UT-B urea transporter in rats fed a low-protein diet (LPD), a high-protein diet (HPD), and a 20% urea-supplemented diet. In the kidney, UT-B protein abundance increased in the outer medulla of both LPD-fed rats and 20% urea-fed rats, without changes in the inner medulla of either group compared with controls. In HPD-fed rats, UT-B protein decreased significantly in both the outer and inner medulla. We identified expression of UT-B in the rat colon, as a 2-kb mRNA transcript and as an approximately 45-kDa protein, with apical localization in superficial colon epithelial cells. UT-B also is expressed in rat small intestine. In rat colon, UT B protein abundance was mildly, but significantly, decreased in LPD-fed and 20% urea-fed rats. UT-B abundance also was examined in the colon of 7/8 nephrectomized, uremic rats and in HPD-fed rats and was not significantly different from that in control rats. These findings indicate that UT-B expression is regulated in response to different loads of urea, with a pattern that suggests involvement of tissue-specific regulatory mechanism in kidney and colon. PMID- 15798088 TI - Rosiglitazone restores renal D1A receptor-Gs protein coupling by reducing receptor hyperphosphorylation in obese rats. AB - Dopamine D(1A) receptor function is impaired in obesity-induced insulin resistance, contributing to sodium retention. We showed previously that uncoupling of D(1A) receptors from G proteins is responsible for diminished natriuretic response to dopamine in obese Zucker rats (OZRs). We hypothesized that overexpression of G protein-coupled receptor kinases (GRKs) leads to increased phosphorylation of D(1A) receptors, which in turn causes uncoupling of the receptors from G(s) proteins in proximal tubules of OZRs. We also examined effects of an insulin sensitizer, rosiglitazone, in correcting these defects. We found that basal and agonist (fenoldopam)-induced coupling of D(1A) receptors to G(s) proteins was impaired in proximal tubules of OZRs compared with lean Zucker rats (LZRs). Moreover, basal serine phosphorylation of D(1A) receptors was elevated two- to threefold in proximal tubules of OZRs compared with LZRs. Fenoldopam increased D(1A) receptor phosphorylation in proximal tubules of LZRs but not OZRs. Compared with that in LZRs, GRK4 expression in OZRs was elevated 200-300% in proximal tubule cell lysates and GRK2 expression was approximately 30% higher in plasma membranes isolated from proximal tubules of OZRs. Rosiglitazone treatment restored basal and agonist-induced coupling of D(1A) receptors to G(s) proteins and reduced basal serine phosphorylation of D(1A) receptors, GRK4 expression, and translocation of GRK2 to the plasma membrane in proximal tubules of OZRs. Furthermore, rosiglitazone significantly reduced fasting blood glucose and plasma insulin in OZRs. Collectively, these results suggest that insulin resistance is responsible for GRK4 overexpression and GRK2 translocation leading to hyperphosphorylation of D(1A) receptors and their uncoupling from G(s) proteins as rosiglitazone treatment corrects these defects in OZRs. PMID- 15798089 TI - Differential renal gene expression in prehypertensive and hypertensive spontaneously hypertensive rats. AB - Development of hypertension stems from both environmental and genetic factors wherein the kidney plays a central role. Spontaneously hypertensive rats (SHR) and the nonhypertensive Wistar-Kyoto (WKY) controls are widely used as a model for studying hypertension. The present study examined the renal gene expression profiles between SHR and WKY at a prehypertensive stage (3 wk of age) and hypertensive stage (9 wk of age). Additionally, age-related changes in gene expression patterns were examined from 3 to 9 wk in both WKY and SHR. Five to six individual kidney samples of the same experimental group were pooled together, and quadruplicate hybridizations were performed using the National Institute of Environmental Health Sciences Rat version 2.0 Chip, which contains approximately 6,700 genes. Twenty two genes were found to be differentially expressed between SHR and WKY at 3 wk of age, and 104 genes were differentially expressed at 9 wk of age. Soluble epoxide hydrolase (Ephx2) was found to be significantly upregulated in SHR at both time points and was the predominant outlier. Conversely, elastase 1 (Ela1) was found to be the predominant gene downregulated in SHR at both time points. Analysis of profiles at 3 vs. 9 wk of age identified 508 differentially expressed genes in WKY rats. In contrast, only 211 genes were found to be differentially expressed during this time period in SHR. The altered gene expression patterns observed in the age-related analysis suggested significant differences in the vascular extracellular matrix system between SHR and WKY kidney. Together, our data highlight the complexity of hypertension and the numerous genes involved in and affected by this condition. PMID- 15798090 TI - Apical ammonia transport by the mouse inner medullary collecting duct cell (mIMCD 3). AB - The collecting duct is the primary site of urinary ammonia secretion; the current study determines whether apical ammonia transport in the mouse inner medullary collecting duct cell (mIMCD-3) occurs via nonionic diffusion or a transporter mediated process and, if the latter, presents the characteristics of this apical ammonia transport. We used confluent cells on permeable support membranes and examined apical uptake of the ammonia analog [(14)C]methylammonia ([(14)C]MA). mIMCD-3 cells exhibited both diffusive and saturable, transporter-mediated, nondiffusive apical [(14)C]MA transport. Transporter-mediated [(14)C]MA uptake had a K(m) of 7.0 +/- 1.5 mM and was competitively inhibited by ammonia with a K(i) of 4.3 +/- 2.0 mM. Transport activity was stimulated by both intracellular acidification and extracellular alkalinization, and it was unaltered by changes in membrane voltage, thereby functionally identifying an apical, electroneutral NH(4)(+)/H(+) exchange activity. Transport was bidirectional, consistent with a role in ammonia secretion. In addition, transport was not altered by Na(+) or K(+) removal, not inhibited by luminal K(+), and not mediated by apical H(+)-K(+) ATPase, Na(+)-K(+)-ATPase, or Na(+)/H(+) exchange. Finally, mIMCD-3 cells express the recently identified ammonia transporter family member Rh C glycoprotein (RhCG) at its apical membrane. These studies indicate that the renal collecting duct cell mIMCD-3 has a novel apical, electroneutral Na(+)- and K(+)-independent NH(4)(+)/H(+) exchange activity, possibly mediated by RhCG, that is likely to mediate important components of collecting duct ammonia secretion. PMID- 15798091 TI - Ischemic proximal tubular injury primes mice to endotoxin-induced TNF-alpha generation and systemic release. AB - Endotoxemia (LPS) can exacerbate ischemic tubular injury and acute renal failure (ARF). The present study tested the following hypothesis: that acute ischemic damage sensitizes the kidney to LPS-mediated TNF-alpha generation, a process that can worsen inflammation and cytotoxicity. CD-1 mice underwent 15 min of unilateral renal ischemia. LPS (10 mg/kg iv), or its vehicle, was injected either 45 min before, or 18 h after, the ischemic event. TNF-alpha responses were gauged 2 h post-LPS injection by measuring plasma/renal cortical TNF-alpha and renal cortical TNF-alpha mRNA. Values were contrasted to those obtained in sham operated mice or in contralateral, nonischemic kidneys. TNF-alpha generation by isolated mouse proximal tubules (PTs), and by cultured proximal tubule (HK-2) cells, in response to hypoxia-reoxygenation (H/R), oxidant stress, antimycin A (AA), or LPS was also assessed. Ischemia-reperfusion (I/R), by itself, did not raise plasma or renal cortical TNF-alpha or its mRNA. However, this same ischemic insult dramatically sensitized mice to LPS-mediated TNF-alpha increases in both plasma and kidney (approximately 2-fold). During late reperfusion, increased TNF alpha mRNA levels also resulted. PTs generated TNF-alpha in response to injury. Neither AA nor LPS alone induced an HK-2 cell TNF-alpha response. However, when present together, AA+LPS induced approximately two- to fivefold increases in TNF alpha/TNF-alpha mRNA. We conclude that modest I/R injury, and in vitro HK-2 cell mitochondrial inhibition (AA), can dramatically sensitize the kidney/PTs to LPS mediated TNF-alpha generation and increases in TNF-alpha mRNA. That ischemia can "prime" tubules to LPS response(s) could have potentially important implications for sepsis syndrome, concomitant renal ischemia, and for the induction of ARF. PMID- 15798092 TI - Analysis of tumor-host interactions by gene expression profiling of lung adenocarcinoma xenografts identifies genes involved in tumor formation. AB - Tumor cell lines are relied on extensively for cancer investigations, yet cultured cells in an in vitro environment differ considerably in behavior compared with those of the same cancer cells that proliferate and form tumors in vivo. To uncover gene expression changes related to tumor formation, gene expression profiles of human lung adenocarcinoma (A549) cells grown as lung tumors in immune-compromised mice were compared with profiles of the same cells grown in vitro. Additionally, profiles of uninvolved adjacent mouse tissue were determined. A profound interplay between cancer cells and the host was shown that affected a complex protein interaction network involving processes of extracellular interaction, growth factor signaling, hemostasis, immune response, and transcriptional regulation. Growth in vivo of A549 cells, which carry an activating k-ras mutation, induced changes in gene expression that corresponded highly to a pattern characteristic of human lung tumors with k-ras mutation. Cytokines interleukin-4, interleukin-6, and IFN-gamma each induced distinct in vitro genomic responses in cancer cells that emulated many of the changes in gene expression observed in vivo. Genes that were both selectively induced in vivo and overexpressed in human lung adenocarcinoma tumors included CSPG2, which has not been associated previously with tumor formation. Knockdown in A549 of CSPG2 by RNA interference significantly inhibited tumor growth in vivo but not in vitro. Thus, analysis of tumor xenografts by gene expression profiling has the potential for identifying genes involved in tumor development that may not be expressed in cancer cells grown in vitro. PMID- 15798093 TI - Components of DNA damage checkpoint pathway regulate UV exposure-dependent alterations of gene expression of FHIT and WWOX at chromosome fragile sites. AB - Common chromosome fragile sites are highly recombinogenic and susceptible to deletions during the development of environmental carcinogen-induced epithelial tumors. Previous studies showed that not only genetic but also epigenetic alterations in cancerous cells are involved in inactivation of the genes FHIT and WWOX at chromosome fragile sites, reported to be potential tumor suppressor genes. Here we investigated the effect of UV light on the gene expression. After exposure to UV, the mRNA and protein of the two genes in murine embryonic fibroblasts (MEF) were unstable, apparently at the G1-S phase of the cell cycle, which was consistent with nuclear run-on assay. A study of MEFs synchronized via a double thymidine block indicated that, after the exposure, the expression of Fhit and Wwox was reduced in E2f-1-deficient cells and markedly in wild-type cells, whereas the reduction was partially inhibited in Trp53-deficient cells; cells at the S phase seemed to be sensitive to exogenous FHIT, suggesting a role of the checkpoint at the G1-S phase in the stability of gene expression and a possible involvement of FHIT function at the S phase. The transfection experiment showed that the UV-induced decrease in expression was partially inhibited by transfection of kinase-dead Atr (ataxia telangiectasia mutated and Rad3 related), which is a sensor of UV-induced damage. Taken together, the present study showed that UV-induced alterations of the fragile site gene expression are involved at least partially in the checkpoint function, suggesting the role in the process of carcinogenesis after exposure to UV. PMID- 15798094 TI - The loss of a single telomere can result in instability of multiple chromosomes in a human tumor cell line. AB - Spontaneous telomere loss has been proposed as an important mechanism for initiating the chromosome instability commonly found in cancer cells. We have previously shown that spontaneous telomere loss in a human cancer cell line initiates breakage/fusion/bridge (B/F/B) cycles that continue for many cell generations, resulting in DNA amplification and translocations on the chromosome that lost its telomere. We have now extended these studies to determine the effect of the loss of a single telomere on the stability of other chromosomes. Our study showed that telomere acquisition during B/F/B cycles occurred mainly through translocations involving either the nonreciprocal transfer or duplication of the arms of other chromosomes. Telomere acquisition also occurred through small duplications involving the subtelomeric region of the other end of the same chromosome. Although all of these mechanisms stabilized the chromosome that lost its telomere, they differed in their consequences for the stability of the genome as a whole. Telomere acquisition involving nonreciprocal translocations resulted in the loss of a telomere on the donor chromosome, which consequently underwent additional translocations, isochromosome formation, or complete loss. In contrast, telomere acquisition involving duplications stabilized the genome, although the large duplications created substantial allelic imbalances. Thus, the loss of a single telomere can generate a variety of chromosome alterations commonly associated with human cancer, not only on a chromosome that loses its telomere but also on other chromosomes. Factors promoting telomere loss are therefore likely to have an important role in generating the karyotype evolution associated with human cancer. PMID- 15798095 TI - Epidermal growth factor induces WISP-2/CCN5 expression in estrogen receptor-alpha positive breast tumor cells through multiple molecular cross-talks. AB - Epidermal growth factor (EGF) is a mitogen for estrogen receptor (ER)-positive breast tumor cells, and it has been proven that EGF occasionally mimicked estrogen action and cross-talks with ER-alpha to exert its activity. Therefore, the present study was undertaken to explore whether EGF is able to modulate the expression of Wnt-1-induced signaling protein-2/connective tissue growth factor/cysteine-rich 61/nephroblastoma overexpressed 5 (WISP-2/CCN5), an estrogen responsive gene, in normal and transformed cell lines of the human breast and, if so, whether this induction is critical for EGF mitogenesis and what downstream signaling pathways are associated with this event. Here, we show that EGF-induced WISP-2 expression in ER- and EGF receptor-positive noninvasive MCF-7 breast tumor cells was dose and time dependent and that expression was modulated at transcription level. A synergism was seen in combination with estrogen. Moreover, small interfering RNA-mediated inhibition of WISP-2/CCN5 activity in MCF-7 cells resulted in abrogation of proliferation by EGF. The multiple molecular cross talks, including the interactions between phosphatidylinositol 3-kinase/Akt and mitogen-activated protein kinase signaling pathways and two diverse receptors (i.e., ER-alpha and EGFR), were essential in the event of EGF-induced WISP-2/CCN5 up-regulation in MCF-7 cells. Moreover, EGF action on WISP-2/CCN5 is restricted to ER- and EGFR-positive noninvasive breast tumor cells, and this effect of EGF cannot be instigated in ER-alpha-negative and EGFR-positive normal or invasive breast tumor cells by introducing ER-alpha. Finally, regulation of phosphorylation of ER-alpha and EGFR may play critical roles in EGF-induced transcriptional activation of WISP-2 gene in breast tumor cells. PMID- 15798096 TI - The progression of LNCaP human prostate cancer cells to androgen independence involves decreased FOXO3a expression and reduced p27KIP1 promoter transactivation. AB - The progression of human prostate cancer from the initial androgen-dependent phase to androgen independence involves diminished apoptosis and a release from the cell cycle block triggered by androgen ablation therapy. FOXO transcription factors play a central role in promoting expression of proapoptotic and cell cycle regulatory genes (e.g., FasL and p27KIP1). Reduced FOXO function might, therefore, play a role in androgen-independent progression of human prostate cancer. Herein, we show that FOXO function is compromised in androgen-independent prostate cancer cells (LNAI) versus androgen-dependent LNCaP cells. The FOXO3a protein, the most highly expressed FOXO family member in prostate cancer cells, is hyperphosphorylated in LNAI cells. FOXO3a expression is also markedly reduced in these androgen-independent LNAI cells when compared with parental LNCaP cells. Together, reduced FOXO3a expression coupled to FOXO3a hyperphosphorylation would suppress FOXO transcriptional activity. Accordingly, activity of the FOXO responsive p27KIP1 promoter is reduced 60% in these LNAI cells when compared with LNCaP cells. Moreover, mutation of a conserved FOXO response element suppresses p27KIP1 promoter activity, substantiating a regulatory role for this FOXO response element in p27KIP1 promoter transactivation. Finally, we show that the activity of a distinct FOXO-responsive promoter, the 3X-IRS promoter, is also reduced in LNAI cells. Collectively, these data show that reduced FOXO3a expression coupled to increased FOXO3a phosphorylation coincide with reduced FOXO responsive promoter activity in androgen-independent LNAI cells when compared with androgen-dependent LNCaP cells. To the extent that this model reflects human disease, these data suggest that FOXO function may be compromised with androgen independent progression of human prostate cancer. PMID- 15798097 TI - Pim-1 kinase stability is regulated by heat shock proteins and the ubiquitin proteasome pathway. AB - Elevated expression of the serine/threonine kinase Pim-1 increases the incidence of lymphomas in Pim-1 transgenic mice and has also been found to occur in some human cancers. Pim-1 acts as a cell survival factor and may prevent apoptosis in malignant cells. It was therefore of interest to understand to what extent maintenance and degradation of Pim-1 protein is affected by heat shock proteins (Hsp) and the ubiquitin-proteasome pathway in K562 and BV173 human leukemic cells. The half-life of Pim-1 protein in these cells was found to increase from 1.7 to 3.1 hours when induced by heat shock or by treating the cells with the proteasome inhibitor PS-341 (bortezomib). The Hsp90 inhibitor geldanamycin prevented the stabilization of Pim-1 by heat shock. Using immunoprecipitation, it was determined that Pim-1 is targeted for degradation by ubiquitin and that Hsp70 is associated with Pim-1 under these circumstances. Conversely, Hsp90 was found to protect Pim-1 from proteasomal degradation. A luminescence-based kinase assay showed that Pim-1 kinase bound to Hsp70 or Hsp90 remains active, emphasizing the importance of its overall cellular levels. This study shows how Pim-1 levels can be modulated in cells through degradation and stabilization. PMID- 15798099 TI - Focus issue: going for the gut. AB - This week's issues of Science and Science's STKE cover various aspects of gastrointestinal function and malfunction. Topics include the regulation of feeding behavior, host interactions with the organisms that colonize the gut, signaling pathways involved in gut barrier function and self-renewal, and factors associated with gastrointestinal inflammatory disorders and the pathogenesis of gastrointestinal cancers. PMID- 15798098 TI - Noncalcemic actions of vitamin D receptor ligands. AB - 1alpha,25-Dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)], the active metabolite of vitamin D(3), is known for the maintenance of mineral homeostasis and normal skeletal architecture. However, apart from these traditional calcium-related actions, 1,25-(OH)(2)D(3) and its synthetic analogs are being increasingly recognized for their potent antiproliferative, prodifferentiative, and immunomodulatory activities. These actions of 1,25-(OH)(2)D(3) are mediated through vitamin D receptor (VDR), which belongs to the superfamily of steroid/thyroid hormone nuclear receptors. Physiological and pharmacological actions of 1,25-(OH)(2)D(3) in various systems, along with the detection of VDR in target cells, have indicated potential therapeutic applications of VDR ligands in inflammation (rheumatoid arthritis, psoriatic arthritis), dermatological indications (psoriasis, actinic keratosis, seborrheic dermatitis, photoaging), osteoporosis (postmenopausal and steroid-induced osteoporosis), cancers (prostate, colon, breast, myelodysplasia, leukemia, head and neck squamous cell carcinoma, and basal cell carcinoma), secondary hyperparathyroidism, and autoimmune diseases (systemic lupus erythematosus, type I diabetes, multiple sclerosis, and organ transplantation). As a result, VDR ligands have been developed for the treatment of psoriasis, osteoporosis, and secondary hyperparathyroidism. Furthermore, encouraging results have been obtained with VDR ligands in clinical trials of prostate cancer and hepatocellular carcinoma. This review deals with the molecular aspects of noncalcemic actions of vitamin D analogs that account for the efficacy of VDR ligands in the above-mentioned indications. PMID- 15798100 TI - Food fight: the NPY-serotonin link between aggression and feeding behavior. AB - Aggressive behavior in mammals is linked to feeding. Interactions between neuropeptide tyrosine (NPY) and serotonergic neurons provide this link. Studies of aggressive behavior in mice in which the Y(1) subtype of NPY receptor was knocked out further define the synaptic circuits responsible for the interrelation of these instinctive behaviors. PMID- 15798101 TI - Signaling the junctions in gut epithelium. AB - This Perspective summarizes recent developments in our understanding of the signaling pathways involved in the regulation of epithelial cell adhesion in the gut. The role of phosphatidylinositol 3-kinase signaling in the modulation of adherens junctions, and the connections between tight junctions and nuclear transcription factors, are discussed. The effect of gastrins on adherens and tight junctions is presented as an example of the regulation of adhesion by growth factors. The consequences of dysregulation of adherens junctions and tight junctions for human pathology are also considered. PMID- 15798102 TI - Orchestration of aberrant epithelial signaling by Helicobacter pylori CagA. AB - Persistent colonization by Helicobacter pylori is the strongest risk factor for distal gastric adenocarcinoma, and H. pylori strains that harbor the cag pathogenicity island further augment cancer risk. The H. pylori cag island encodes a secretion system, and the product of the terminal gene in the island (CagA) is translocated into host epithelial cells after bacterial attachment, where it undergoes tyrosine phosphorylation by Src kinases and binds the eukaryotic phosphatase SHP-2. Higashi et al. now demonstrate that CagA-dependent SHP-2 activation leads to sustained activation of ERK (extracellular signal regulated kinase), culminating in morphological changes that mimic unrestrained stimulation by growth factors. These data implicate the cag island as a key mediator of pathogenic epithelial responses that may heighten the risk for gastric cancer. PMID- 15798103 TI - Central and peripheral signaling mechanisms involved in endocannabinoid regulation of feeding: a perspective on the munchies. AB - The endocannabinoid system is a critical regulator of energy homeostasis and food intake. Through cannabinoid (CB)(1) receptors in the brain and periphery, endocannabinoids exert powerful effects on the systems of the body that coordinate the balance between food intake, metabolism, and energy expenditure. These integrative systems control food intake both by modulating the inputs to various brain areas that monitor energy balance and by increasing the hedonic or reward value of the food consumed. Cannabinoids also alter metabolism, acting through both centrally located CB(1) receptors that drive neuronal pathways controlling metabolism and peripheral CB(1) receptors located in tissues throughout the body. PMID- 15798105 TI - Lack of occupational health research in Nepal. PMID- 15798106 TI - Addressing social determinants of health inequities: learning from doing. PMID- 15798107 TI - Using surveillance data to develop and disseminate local childhood lead poisoning screening recommendations: Miami-Dade County's experience. AB - Before 1999, few children in Florida's Miami-Dade County were being screened for lead poisoning. To improve screening rates, the county's department of health developed screening recommendations and a screening tool using surveillance and census data and disseminated these materials to primary care providers. Each year, recommendations have been reviewed to assess their sensitivity, and revised recommendations have been disseminated to health care providers. The percentage of children 6 years or younger screened in Florida who reside in Miami-Dade County increased from 4.1% in 1998 to 20.3% in 2002. Analysis and dissemination of blood lead surveillance data not only guide development of screening recommendations but also educate health care providers regarding the importance of childhood screening. PMID- 15798108 TI - Faces of change. PMID- 15798109 TI - From neurons to King County neighborhoods: partnering to promote policies based on the science of early childhood development. AB - A diverse community partnership in the Seattle area developed a policy agenda based on science, organized community support, and committed to monitor policy changes. It found that public health agencies are well positioned to develop a common knowledge base on early childhood development and to initiate community coalitions promoting policies to strengthen environments. It was challenging to maintain participants' focus on environments over time. Providing access to conditions that promote optimal development for all children will require ongoing commitment and alignment of many sectors to move political will and mobilize for change. PMID- 15798110 TI - Talking about public health: developing America's "second language". AB - The mission of public health--improving the health of populations--is difficult to advance in public discourse because a language to express the values animating that mission has not been adequately developed. Following on the work of Robert Bellah, Dan Beauchamp, and others, we argue that the first "language" of American culture is individualism. A second American language of community--rooted in egalitarianism, humanitarianism, and human interconnection--serves as the first language of public health. These values resonate with many Americans but are not easily articulated. Consequently, reductionist, individualistic understandings of public health problems prevail. Advancing the public health approach to the nation's health challenges requires invigorating America's second language by recognizing the human interconnection underlying the core social justice values of public health. PMID- 15798112 TI - Jacobson v Massachusetts at 100 years: police power and civil liberties in tension. AB - A century ago, the US Supreme Court in Jacobson v Massachusetts upheld the exercise of the police power to protect the public's health. Despite intervening scientific and legal advances, public health practitioners still struggle with Jacobson's basic tension between individual liberty and the common good. In affirming Massachusetts' compulsory vaccination law, the Court established a floor of constitutional protections that consists of 4 standards: necessity, reasonable means, proportionality, and harm avoidance. Under Jacobson, the courts are to support public health matters insofar as these standards are respected. If the Court today were to decide Jacobson once again, the analysis would likely differ--to account for developments in constitutional law--but the outcome would certainly reaffirm the basic power of government to safeguard the public's health. PMID- 15798111 TI - Manifold restraints: liberty, public health, and the legacy of Jacobson v Massachusetts. AB - February 2005 marks the centenary of one of the most important pieces of public health jurisprudence, the US Supreme Court case of Jacobson v Massachusetts, which upheld the authority of states to pass compulsory vaccination laws. The Court's decision articulated the view that the freedom of the individual must sometimes be subordinated to the common welfare. We examined the relationship between the individual and society in 20th-century public health practice and law and the ways that compulsory measures have been used to constrain personal liberty for the sake of protecting the public health. PMID- 15798113 TI - Jacobson v Massachusetts: it's not your great-great-grandfather's public health law. AB - Jacobson v Massachusetts, a 1905 US Supreme Court decision, raised questions about the power of state government to protect the public's health and the Constitution's protection of personal liberty. We examined conceptions about state power and personal liberty in Jacobson and later cases that expanded, superseded, or even ignored those ideas. Public health and constitutional law have evolved to better protect both health and human rights. States' sovereign power to make laws of all kinds has not changed in the past century. What has changed is the Court's recognition of the importance of individual liberty and how it limits that power. Preserving the public's health in the 21st century requires preserving respect for personal liberty. PMID- 15798114 TI - Integrating social theory into public health practice. AB - The innovative practice that resulted from the Ottawa Charter challenges public health knowledge about programming and evaluation. Specifically, there is a need to formulate program theory that embraces social determinants of health and local actors' mobilization for social change. Likewise, it is imperative to develop a theory of evaluation that fosters reflexive understanding of public health programs engaged in social change. We believe advances in contemporary social theory that are founded on a critique of modernity and that articulate a coherent theory of practice should be considered when addressing these critical challenges. PMID- 15798115 TI - Building on leadership and social capital to create change in 2 urban communities. AB - There has been an appeal to reduce health inequities by increasing community involvement and social capital. Poder es Salud/Power for Health is a community based participatory prevention research project that seeks to address health disparities in the African American and Latino communities by enhancing community level social capital. We provide specific examples of how this intervention uses community health workers and popular education to reduce language and cultural barriers and enhance community social capital. Although the communities share fundamental challenges related to health disparities, the ways in which the Latino and African American communities identify health concerns, create solutions, and think about social capital vary. Members of the project are working together to identify opportunities for cross-cultural collaboration. PMID- 15798116 TI - Becoming the Framingham Study 1947-1950. AB - In the epidemiological imagination, the Framingham Heart Study has attained iconic status, both as the prototype of the cohort study and as a result of its scientific success. When the Public Health Service launched the study in 1947, epidemiological knowledge of coronary heart disease was poor, and epidemiology primarily involved the study of infectious disease. In constructing their investigation, Framingham's initiators had to invent new approaches to epidemiological research. These scientific goals were heavily influenced by the contending institutional and personal interests buffeting the study. The study passed through vicissitudes and stages during its earliest years as its organizers grappled to define its relationship to medicine, epidemiology, and the local community. PMID- 15798117 TI - The community action model: a community-driven model designed to address disparities in health. AB - The community action model is a 5-step, community-driven model designed to build communities' capacity to address health disparities through mobilization. Fundamental to the model is a critical analysis identifying the underlying social, economic, and environmental forces that create health and social inequities in a community. The goal is to provide communities with the framework necessary to acquire the skills and resources to plan, implement, and evaluate health-related actions and policies. The model was developed in the context of tobacco-related health disparities. Concrete policy outcomes demonstrate the model's potential application to a wide variety of grassroots policy development efforts. PMID- 15798118 TI - "Come and take a walk": listening to Early Head Start parents on school-readiness as a matter of child, family, and community health. AB - We explored the perspectives and experiences of low-income, predominantly African American families regarding children's school-readiness. Our research, which involved qualitative interviews, ethnographic case studies, and "photovoice" methods, focused on families participating in the national evaluation of Early Head Start. While valuing academic skills, study parents emphasized the importance of social and emotional health in regard to both children's and parents' readiness to begin school. These developments are especially critical given the challenges parents perceive in local school environments. On the basis of a social ecology framework, we argue that psychological and environmental dimensions of school-readiness are public health matters and that understanding the perspectives of low-income and minority parents on such issues is a critical aspect of health communication dedicated to eliminating health disparities. PMID- 15798119 TI - Tackling the wider determinants of health disparities in England: a model for evaluating the New Deal for Communities regeneration initiative. AB - The National Strategy for Neighbourhood Renewal (NSNR) is a central component of British policy to reduce health disparities. This program seeks to improve local socioeconomic and physical environments through the intensive regeneration of disadvantaged communities. We describe the challenges facing evaluators tasked with assessing the impacts of 1 component of the NSNR--the New Deal for Communities initiative--and explore techniques that may be adopted in the evaluation process. PMID- 15798120 TI - Adult socioeconomic position and the association between height and coronary heart disease mortality: findings from 33 years of follow-up in the Whitehall Study. AB - In the Whitehall Study, which followed 17,139 male civil servants over 33 years, the association between tall stature and coronary heart disease (CHD) mortality differed between employment grades. In men without CHD at study entry, the hazard ratio per 15-cm increase in height was 0.77 (95% confidence interval [CI]=0.69, 0.85; P<0.001) for the highest grades, but 0.84 (95% CI=0.69, 1.03; P=.10) for middle and 0.95 (95% CI=0.75, 1.20; P=.65) for low grades, suggesting that childhood and adult social conditions may interact in their influence on coronary risk. PMID- 15798121 TI - The obesity epidemic as harbinger of a metabolic disorder epidemic: trends in overweight, hypercholesterolemia, and diabetes treatment in Geneva, Switzerland, 1993-2003. AB - Increases in obesity, hypercholesterolemia, and diabetes may be under way in Europe. We have reported the only data available from the 1990s for continuous monitoring of chronic disease risk factors in random samples of a general European population. In random surveys (1993-2003) of 6164 men and 6107 women in Geneva, overweight and obesity combined increased in both men and women; hypercholesterolemia prevalence also rose; diabetes treatment increased in men. Only population-based interventions can prevent the impending epidemic of obesity related disorders. PMID- 15798122 TI - Heat-related fatalities in North Carolina. AB - We used medical examiner's records to identify heat-related fatalities (N=161) that occurred during the period January 1, 1977, to December 31, 2001, in North Carolina. Estimates of the population at risk were derived from US census data. Annual fatality rates increased with increases in average summer temperature and with the number of days per year at 90 degrees F or higher. Of the occupational heat-related fatalities (n=40), 45% occurred among farm laborers, many of whom died unnoticed and without medical attention. PMID- 15798123 TI - Immigration and geographic access to prenatal clinics in Brooklyn, NY: a geographic information systems analysis. AB - We compared levels of geographic access to prenatal clinics in Brooklyn, NY, between immigrant and US-born mothers and among immigrant groups by country of birth. We used birth data to characterize the spatial distribution of mothers and kernel estimation to measure clinic density within a 2-mile radius of each mother. Results showed that geographic access to clinics differs substantially by country of birth. Certain groups (e.g., Pakistani, Bangladeshi) have relatively poor geographic access despite a high need for prenatal care. PMID- 15798124 TI - The National Public Health Leadership Institute: evaluation of a team-based approach to developing collaborative public health leaders. AB - Recent public health literature contains calls for collaborative public health interventions and for leaders capable of guiding them. The National Public Health Leadership Institute aims to develop collaborative leaders and to strengthen networks of leaders who share knowledge and jointly address public health problems. Evaluation results show that completing the institute training increases collaborative leadership and builds knowledge-sharing and problem solving networks. These practices and networks strengthen interorganizational relationships, coalitions, services, programs, and policies. Intensive team-and project-based learning are key to the program's impact. PMID- 15798125 TI - Healthy eating and exercising to reduce diabetes: exploring the potential of social determinants of health frameworks within the context of community-based participatory diabetes prevention. AB - OBJECTIVES: We examined a community-based participatory diabetes intervention to identify facilitators of and barriers to sustained community efforts to address social factors that contribute to health. METHODS: We conducted a case study description and analysis of the Healthy Eating and Exercising to Reduce Diabetes project in the theoretical context of a conceptual model of social determinants of health. RESULTS: We identified several barriers to and facilitators of analysis of social determinants of a community-identified disease priority (in this case, diabetes). Barriers included prevailing conceptual models, which emphasize health behavioral and biomedical paradigms that exclude social determinants of health. Facilitating factors included (1) opportunities to link individual health concerns to social contexts and (2) availability of support from diverse partners with a range of complementary resources. CONCLUSIONS: Partnerships that offer community members tangible resources with which to manage existing health concerns and that integrate an analysis of social determinants of health can facilitate sustained engagement of community members and health professionals in multilevel efforts to address health disparities. PMID- 15798126 TI - The Seattle-King County Healthy Homes Project: a randomized, controlled trial of a community health worker intervention to decrease exposure to indoor asthma triggers. AB - OBJECTIVES: We assessed the effectiveness of a community health worker intervention focused on reducing exposure to indoor asthma triggers. METHODS: We conducted a randomized controlled trial with 1-year follow-up among 274 low income households containing a child aged 4-12 years who had asthma. Community health workers provided in-home environmental assessments, education, support for behavior change, and resources. Participants were assigned to either a high intensity group receiving 7 visits and a full set of resources or a low-intensity group receiving a single visit and limited resources. RESULTS: The high-intensity group improved significantly more than the low-intensity group in its pediatric asthma caregiver quality-of-life score (P=.005) and asthma-related urgent health services use (P=.026). Asthma symptom days declined more in the high-intensity group, although the across-group difference did not reach statistical significance (P=.138). Participant actions to reduce triggers generally increased in the high-intensity group. The projected 4-year net savings per participant among the high-intensity group relative to the low-intensity group were 189-721 dollars. CONCLUSIONS: Community health workers reduced asthma symptom days and urgent health services use while improving caregiver quality-of-life score. Improvement was greater with a higher-intensity intervention. PMID- 15798127 TI - Neighborhood racial composition, neighborhood poverty, and the spatial accessibility of supermarkets in metropolitan Detroit. AB - OBJECTIVES: We evaluated the spatial accessibility of large "chain" supermarkets in relation to neighborhood racial composition and poverty. METHODS: We used a geographic information system to measure Manhattan block distance to the nearest supermarket for 869 neighborhoods (census tracts) in metropolitan Detroit. We constructed moving average spatial regression models to adjust for spatial autocorrelation and to test for the effect of modification of percentage African American and percentage poor on distance to the nearest supermarket. RESULTS: Distance to the nearest supermarket was similar among the least impoverished neighborhoods, regardless of racial composition. Among the most impoverished neighborhoods, however, neighborhoods in which African Americans resided were, on average, 1.1 miles further from the nearest supermarket than were White neighborhoods. CONCLUSIONS: Racial residential segregation disproportionately places African Americans in more-impoverished neighborhoods in Detroit and consequently reduces access to supermarkets. However, supermarkets have opened or remained open close to middle-income neighborhoods that have transitioned from White to African American. Development of economically disadvantaged African American neighborhoods is critical to effectively prevent diet-related diseases among this population. PMID- 15798128 TI - African Americans' access to healthy food options in South Los Angeles restaurants. AB - OBJECTIVES: We examined availability and food options at restaurants in less affluent (target area) and more affluent (comparison area) areas of Los Angeles County to compare residents' access to healthy meals prepared and purchased away from home. We also considered environmental prompts that encourage the purchase of various foods. METHODS: We designed an instrument to assess the availability, quality, and preparation of food in restaurants. We also assessed advertisements and promotions, cleanliness, and service for each restaurant. We assessed 659 restaurants: 348 in the target area and 311 in the comparison area. RESULTS: The nutritional resource environment in our target area makes it challenging for residents to eat healthy away from home. Poorer neighborhoods with a higher proportion of African American residents have fewer healthy options available, both in food selections and in food preparation; restaurants in these neighborhoods heavily promote unhealthy food options to residents. CONCLUSIONS: Environment is important in understanding health status: support for the healthy lifestyle associated with lower risks for disease is difficult in poorer communities with a higher proportion of African American residents. PMID- 15798129 TI - Primary care, social inequalities, and all-cause, heart disease, and cancer mortality in US counties, 1990. AB - OBJECTIVES: We tested the association between the availability of primary care and income inequality on several categories of mortality in US counties. METHODS: We used cross-sectional analysis of data from counties (n=3081) in 1990, including analysis of variance and multivariate ordinary least squares regression. Independent variables included primary care resources, income inequality, and sociodemographics. RESULTS: Counties with higher availability of primary care resources experienced between 2% and 3% lower mortality than counties with less primary care. Counties with high income inequality experienced between 11% and 13% higher mortality than counties with less inequality. CONCLUSIONS: Primary care resources may partially moderate the effects of income inequality on health outcomes at the county level. PMID- 15798130 TI - Father's occupational group and daily smoking during adolescence: patterns and predictors. AB - OBJECTIVES: We investigated the relationship among father's occupational group, daily smoking, and smoking determinants in a cohort of New Zealand adolescents. METHODS: The longitudinal Multidisciplinary Health and Development Study provided information on adolescents' self-reported smoking behavior and potential predictors of smoking, such as social and material factors, personality characteristics, educational achievement, and individual attitudes and beliefs regarding smoking. Longitudinal logistic generalized estimating equation analyses were used. RESULTS: Adolescents whose fathers were classified in the lowest status occupational group were twice as likely as those whose fathers occupied the highest-status occupational group to be daily smokers. This high risk of daily smoking among the adolescents from the lowest occupational group was largely predicted by their lower intelligence scores and by the higher prevalence of smoking among fathers and friends. CONCLUSIONS: To prevent socioeconomic differences in smoking, school-based interventions should seek to prevent smoking uptake among adolescents, particularly those of lower socioeconomic status. Programs need to provide positive, nonsmoking role models consonant with the culture and norms of lower-socioeconomic-status groups. Adolescents need to acquire resistance skills and protective behaviors against social pressure and influences. PMID- 15798132 TI - The epidemiology of alcohol, marijuana, and cocaine use among Mexican American, Puerto Rican, Cuban American, and other Latin American eighth-grade students in the United States: 1991-2002. AB - OBJECTIVES: We compared trends in and correlates of marijuana use, cocaine use, and heavy alcohol use for adolescents of Mexican American, Puerto Rican, Cuban, and other Latin American heritage in the United States. METHODS: We used/examined data from nationally representative samples of eighth-grade Hispanic students who participated in the Monitoring the Future study during the years 1991-2002 (n=24235). RESULTS: Drug use was significantly higher among boys and adolescents of almost all Hispanic ethnicities who did not live with both parents. In addition, drug use differed considerably according to ethnic group on language first spoken, parental education, urbanicity, and region. CONCLUSIONS: A better understanding of the homogeneity and heterogeneity of drug use patterns within and between Hispanic groups should assist in the development of prevention programs. PMID- 15798131 TI - Effects of race, neighborhood, and social network on age at initiation of injection drug use. AB - OBJECTIVES: We investigated individual- and neighborhood-level factors associated with adolescent initiation of injection drug use. METHODS: Injection drug users (IDUs) who had been injecting 2 to 5 years underwent HIV testing and completed a sociobehavioral risk survey. Modeling techniques accounting for intraneighborhood correlations were used in data analyses. RESULTS: Adolescent-initiating IDUs were less likely than adult-initiating IDUs to report high-risk sex and injection behaviors and more likely to report high-risk networks. African American IDUs from neighborhoods with large percentages of minority residents and low adult educational levels were more likely to initiate injection during adolescence than White IDUs from neighborhoods with low percentages of minority residents and high adult education levels. CONCLUSIONS: Racial segregation and neighborhood-level educational attainment must be considered when drawing inferences about age at initiation of injection drug use and related high-risk behaviors. PMID- 15798134 TI - Racial/ethnic variation in functional and self-reported health. AB - OBJECTIVES: We investigated whether racial/ethnic health disparities exist in Canada and whether socioeconomic or behavioral differences between racial/ ethnic minorities and nonminorities account for such disparities. METHODS: We used data from the National Population Health Survey, conducted by Statistics Canada in 1996 and 1997. We used regression models to examine differences in functional and self-reported health. RESULTS: Our study found no association between socioeconomic or behavioral differences and racial/ethnic health disparities. There was no clear pattern between racial/ethnic minority status and health. CONCLUSIONS: The state can play an important role in health outcomes, and public commitment to accessible health care may explain why socioeconomic status and health behaviors are weak indicators of racial/ethnic health variation in Canada. PMID- 15798133 TI - Childhood sexual abuse and age at initiation of injection drug use. AB - OBJECTIVES: We examined the relation between childhood sexual abuse and injection drug use initiation among young adult injection drug users. METHODS: We used mixed effect linear models to compare age at first injection among 2143 young injection drug users by first sexual abuse age categories. RESULTS: The participants were predominantly male (63.3%) and White (52.8%). Mean age and age at first injection were 23.7 and 19.6 years, respectively; 307 participants (14.3%) reported childhood sexual abuse. After adjustment for gender, race/ethnicity, noninjection drug use before first injection drug use, and recruitment site, childhood sexual abuse was independently associated with younger age at first injection. CONCLUSIONS: Childhood sexual abuse was associated with earlier initiation of injection drug use. These data emphasize the need to integrate substance abuse prevention with postvictimization services for children and adolescents. PMID- 15798135 TI - Asthma, suicidal ideation, and suicide attempts: findings from the Baltimore epidemiologic catchment area follow-up. AB - OBJECTIVES: We examined cross-sectional and longitudinal associations between asthma, suicidal ideation, and suicide attempt among adults in the community. METHODS: Data were drawn from 3 waves (1981,1982,1993-1996) of the Baltimore follow-up of the Epidemiologic Catchment Area study. Multiple logistic regression analyses were used to examine associations between asthma, asthma treatment, suicidal ideation, suicide attempt, and suicide completion. RESULTS: Asthma at wave 1 was associated with a significantly increased odds of suicidal ideation (odds ratio [OR] = 2.33; confidence interval [CI] = 1.03, 5.25) and suicide attempt (OR=3.54; CI=1.4, 8.99), which persisted independent of lifetime National Institute of Mental Health Diagnostic Interview Schedule/Diagnostic and Statistical Manual of Mental Disorders, Third Edition major depression and treatment for asthma at wave 2. CONCLUSIONS: These findings provide preliminary evidence suggestive of an association between asthma and an increased likelihood of suicidal ideation and suicide attempt among adults in the community. Neither lifetime major depression nor treatment for asthma explained this relation. These results provide important directions for future research, and if replicated these data may have clinical and public health implications. PMID- 15798136 TI - Depression treatment in primary care. AB - BACKGROUND: Depression costs the United States dollars 40 billion annually. Primary care physicians play a key role in the identification and treatment of depression. This study focused on the treatment options recommended by physicians and whether physicians were following the recommended treatment guidelines. METHODS: We recorded treatment recommendations by examining charts for all patients with newly detected depression. The patients were from 44 family medicine practitioners and 23 general internal medicine practitioners in a Midwest university medical center setting. RESULTS: For both medical specialties combined, pharmacotherapy was the most widely used intervention (recommended for 52% of patients), whereas psychotherapy alone was the least frequently used intervention (recommended for 4% of patients). Family medicine practitioners recommended combination treatment (pharmacotherapy and psychotherapy) more frequently than did general internal medicine practitioners (P = .022), and female physicians recommended combination treatment more frequently than did male physicians (P = .010). CONCLUSIONS: Pharmacotherapy was found to be the most widely used treatment despite current evidence-based recommendations. Barriers to effective treatment plan are discussed. The implications for mental health interventions, combination therapy, and cost offset are also discussed. Further research exploring the negotiation process during the patient-provider encounter would shed light on patient and physician factors influencing treatment decisions. PMID- 15798137 TI - Adherence to mental health treatment in a primary care clinic. AB - BACKGROUND: Patient nonadherence is common for the standard mental health treatments in primary care: antidepressants and referrals to specialty mental health treatment. This is one of few studies to prospectively identify predictors of nonadherence. METHODS: We observed 95 veterans attending an internal medicine clinic prescribed antidepressant medication or referred to mental health treatment. We collected information on sociodemographic factors, health beliefs, preferences about treatment, past experiences, and treatment knowledge. RESULTS: At 1 month, medication adherence was greater when patients experienced previous pharmacy trouble and traveled for less than 30 minutes to reach the clinic. Appointment attendance improved when patients were ready for treatment, perceived benefits, and saw their physician as collaborative. At 6 months, medication adherence was greater when patients reported a preference for medicine treatment, traveled for less than 30 minutes, and perceived greater benefits. Fewer negative effects from previous mental health treatment improved adherence to appointments. In multivariate analyses examining adherence to all treatments, greater readiness for treatment predicted 1-month adherence, whereas being unmarried and seeing the physician as more collaborative improved 6-month adherence. CONCLUSIONS: Adherence to antidepressant medications and to mental health referrals should be examined separately. A brief initial assessment for nonadherence risk factors may identify persons for targeted adherence promoting interventions. PMID- 15798138 TI - Heart rate variability and obesity indices: emphasis on the response to noise and standing. AB - BACKGROUND: Obese people have a higher prevalence of cardiovascular disease, but the mechanism of this result remains obscure. The purpose of this study was to prove heart rate variability (HRV) response at rest and during stimuli in these persons. METHODS: The subjects were 41 healthy persons (19 men, 22 women) ranging in age from 20 to 65 years. HRV was measured at rest and at given stresses with noise and standing. RESULTS: Higher levels of fat mass, percentage fat content, and waist/hip ratio were significantly associated with lower low frequency (LF) (r = -0.34, r = -0.43; P < .01, r = -0.33, P < .05), and lower root mean square differences of successive NN intervals (RMS standard deviation) (r = -0.33, r = 0.35, r = -0.38, P < .05). During rest, noise, and standing, the change amount of the standard deviation of NN interval (SDNN) and low frequency/high frequency ratio were not different between normal and obese groups (P > .05). CONCLUSION: Although there was no significant HRV response to stimuli, root mean square of successive differences (which reflects parasympathetic acivity) and low frequency (which mainly reflects sympathetic activity) were negatively correlated with fat mass, fat percentage, and waist-to-hip ratio at rest in obese persons. These results mean obesity can change cardiac autonomic nervous response, meaning that the mechanism by which obesity increases cardiac mortality would be explained, at least partially. PMID- 15798139 TI - Are family physicians appropriately screening for postpartum depression? AB - PURPOSE: Despite the availability of screening tools for postpartum depression (PPD), there is a general consensus that the condition is underdiagnosed. This study was conducted to determine how frequently family physicians screen for PPD, what methods they use to screen, and what influences their screening frequency. METHODS: A survey of members of the Washington Academy of Family Physicians was conducted. Three hundred sixty-two (60.9%) mailed surveys were returned. The 298 physicians who saw postpartum women and children younger than 1 year of age were included in the study. RESULTS: Of the study population, 70.2% always or often screened for PPD at postpartum gynecologic examinations, and 46% always or often screened mothers at well-child visits. Of those who screened, 30.6% reported using a validated screening tool. Of those, only 18% used a tool specifically designed to screen for PPD. Logistic regression modeling showed that female sex [odds ratio (OR) = 2.2], training in PPD during residency (OR = 8.1), training in PPD through medical literature (OR = 2.1), and agreement that postpartum depression is common enough to warrant screening (OR = 1.9) were all significantly associated with more frequent screening at postpartum gynecologic visits. Agreement that screening takes too much effort was associated with less frequent screening (OR = 0.8). CONCLUSIONS: Although family physicians believe that PPD is serious, identifiable, and treatable; screening is not universal and use of screening tools designed for PPD is uncommon. Training in postpartum depression and female sex are the variables most strongly associated with frequent screening. PMID- 15798140 TI - Fluid matters in choosing antihypertensive therapy: a hypothesis that the data speak volumes. AB - Assuming that blood pressure is lowered equivalently, diuretics are more effective than angiotensin-converting enzyme inhibitors (ACEIs), calcium channel blockers (CCBs), and alpha-adrenergic receptor blockers (alpha-blockers) at preventing heart failure, and they are more effective than ACEIs and alpha blockers at preventing strokes. Compared with beta-adrenergic receptor blockers (beta-blockers) and ACEIs, CCBs are less effective at reducing myocardial infarcts and heart failure. There is currently no conceptual framework by which to organize data indicating that some antihypertensive medications are better than others at preventing cardiovascular diseases. The thesis of this article is that the fluid reduction or fluid retention attributable to antihypertensive medications, either alone or in combination, provides a basis for ranking these medications. Diuretics have a theoretical advantage compared with other antihypertensive medications because they reduce total body fluid more than other agents. Therefore, they are the preferred drugs for treating hypertension. The other antihypertensive agents that promote fluid reduction, ACEIs and angiotensin receptor blockers (ARBs), are next in preference, ranking a close second to diuretics. Because beta-blockers have a neutral effect on total body fluid, they rank on a third tier of preference, after ACEIs and ARBs. CCBs and alpha-blockers are the least preferred medications for treating hypertension because they promote fluid retention. PMID- 15798141 TI - Barriers to the diagnosis and treatment of depression in Jordan. A nationwide qualitative study. AB - BACKGROUND: Depression is one of the most common causes of morbidity in developing countries. It is believed that there are many barriers to diagnosis and treatment in the primary care setting, but little research exists. METHODS: Five focus groups were conducted with the goal of exploring themes related to barriers to the diagnosis and treatment of depression, with a purposeful nationwide sample of 50 primary health care providers working in the public health clinics of the Jordanian Ministry of Health (MOH). Participant comments were transcribed and analyzed by the authors, who agreed on common themes. RESULTS: Lack of education about depression, lack of availability of appropriate therapies, competing clinical demands, social issues, and the lack of patient acceptance of the diagnosis were felt to be among the most important barriers to the identification, diagnosis, and treatment of patients with depression in this population. CONCLUSIONS: Continuing medical education for providers about depression, provision of counseling services and antidepressant medications at the primary care level, and efforts to destigmatize depression may result in increased rates of recognition and treatment of depression in this population. Systematizing traditional social support behaviors may be effective in reducing the numbers of patients referred for medical care. PMID- 15798142 TI - Unsuspected Klinefelter syndrome diagnosed during oncologic evaluation: a case series. AB - Klinefelter syndrome is an underdiagnosed chromosomal disorder that has significant implications for health and for medical management. This report presents 5 adult male patients diagnosed with previously unsuspected Klinefelter syndrome as a result of cytogenetic testing for suspected hematologic malignancies. This case series highlights the importance of maintaining a comprehensive and holistic approach to medical care. The medical, genetic, and psychosocial implications of the Klinefelter diagnosis are discussed. PMID- 15798143 TI - Churg-Strauss syndrome associated with HIV infection. AB - A 28-year-old woman with a history of asthma and recent deep venous thrombosis presented with fever, chest pain, and peripheral eosinophilia. The patient was subsequently diagnosed with Churg-Strauss syndrome and HIV infection, representing to our knowledge only the second case of this association. Rheumatologic manifestations of HIV may precede clinical signs of infection. This is significant because steroidal and cytotoxic therapy may potentially worsen HIV infection. As the prevalence of HIV infection rises, there may be atypical presentations of various rheumatologic syndromes. The following case demonstrates a patient whose initial presentation for HIV infection was Churg-Strauss syndrome. PMID- 15798144 TI - Anaphylaxis induced by glucocorticoids. AB - Glucocorticoids are frequently used to treat allergic reactions. Therefore, allergic reactions to systemic glucocorticoids in particular are considered most unlikely and are not well known. We report on a 23-year-old woman with atopic dermatitis who had an anaphylactic reaction after oral administration of prednisolone. On treatment with epinephrine, antihistamines and volume symptoms resolved. Skin testing with a panel of glucocorticoids showed immediate type reactions to prednisolone, prednisolone hydrogen succinate, prednisone, and betamethasone dihydrogen phosphate. In challenge testing the patient tolerated methyl prednisolone and dexamethasone. There is increasing evidence that true allergic immediate type reactions to glucocorticoids exist. The severity of the reaction can vary from a rash to anaphylaxis. However, a patient sensitized to one or a group of glucocorticoids does not have to refrain from all types of glucocorticoids. Careful challenge testing is by far the best way to select glucocorticoids that are safe for future treatment. Clinicians should be aware that allergic reactions to glucocorticoids can occur and that worsening of symptoms does not always mean treatment failure. PMID- 15798145 TI - Excess factor VIII: a common cause of hypercoagulability. AB - BACKGROUND: Elevations of coagulation factor VIII have recently been described in significant numbers of patients with venous thromboembolism (VTE) previously thought to have had an "idiopathic" event. Three patients from a family practice are presented with recurrent VTE and increased factor VIII levels. METHOD: The case histories and results of laboratory tests were reviewed. RESULTS: The 3 patients had no other clear coagulopathy. In 2 of the 3, the increase was confirmed with a second reading. CONCLUSIONS: Factor VIII assay should be considered in the work-up of idiopathic, recurrent VTE. Long-term anticoagulation may be appropriate in this setting. PMID- 15798147 TI - Colonoscopy in rural communities. PMID- 15798148 TI - Repeating failed policies will not improve outcomes. PMID- 15798152 TI - Quantitative neuropathologic correlates of changes in ratio of N-acetylaspartate to creatine in macaque brain. AB - PURPOSE: To elucidate the neuropathologic basis of transient changes in the ratio of N-acetylaspartate (NAA) to creatine (Cr) in the primate brain by using a simian immunodeficiency virus (SIV)-infected macaque model of the neurologic manifestation of acquired immune deficiency syndrome. MATERIALS AND METHODS: This study was approved by the Massachusetts General Hospital Subcommittee on Research and Animal Care and the Institutional Animal Care and Use Committee of Harvard University. Rhesus macaques infected with SIV were evaluated during the 1st month of infection. A total of 11 animals were studied, including four control animals, three animals sacrificed 12 days after infection, three animals sacrificed 14 days after infection, and one animal sacrificed 28 days after infection. All animals underwent in vivo proton ((1)H) magnetic resonance (MR) spectroscopy, and postmortem frontal lobe tissue was investigated by using high-spectral-resolution (1)H MR spectroscopy of brain extracts. In addition, quantitative neuropathologic analyses were performed. Stereologic analysis was performed to determine neuronal counts, and immunohistochemical analysis was performed to analyze three neuronal markers: synaptophysin, microtubule-associated protein 2 (MAP2), and calbindin. Analysis of variance (ANOVA) was used to determine substantial changes in neuropathologic and MR spectroscopic markers. Spearman rank correlations were calculated between plasma viral load and neuropathologic and spectroscopic markers. RESULTS: During acute infection with SIV, the macaque brain exhibited significant changes in NAA/Cr (P < .02, ANOVA) and synaptophysin (P < .013, ANOVA). There was no significant change in the concentration of Cr. No significant changes were found in neuronal counts or other immunohistochemical neuronal markers. With the Spearman rank test, a significant direct correlation was detected between synaptophysin and ex vivo NAA/Cr (r(s) = 0.72, P < .013). No correlation between NAA/Cr and neuronal counts, calbindin, or MAP2 was found. CONCLUSION: NAA/Cr is a sensitive marker of neuronal injury, not necessarily neuronal loss, and best correlates with synaptophysin, a marker of synaptodendritic dysfunction. PMID- 15798153 TI - Current realities of delivering mammography services in the community: do challenges with staffing and scheduling exist? AB - PURPOSE: To evaluate the current (2001-2002) capacity of community-based mammography facilities to deliver screening and diagnostic services in the United States. MATERIALS AND METHODS: Institutional review board approvals and patient consent were obtained. A mailed survey was sent to 53 eligible mammography facilities in three states (Washington, New Hampshire, and Colorado). Survey questions assessed equipment and staffing availability, as well as appointment waiting times for screening and diagnostic mammography services. Criterion related content and construct validity were obtained first by means of a national advisory committee of academic, scientific, and clinical colleagues in mammography that reviewed literature on existing surveys and second by pilot testing a series of draft surveys among community mammography facilities not inclusive of the study facilities. The final survey results were independently double entered into a relational database with programmed data checks. The data were sent encrypted by means of file transfer protocol to a central analytical center at Group Health Cooperative. A two-sided P value with alpha = .05 was considered to show statistical significance in all analyses. RESULTS: Forty-five of 53 eligible mammography facilities (85%) returned the survey. Shortages of radiologists relative to the mammographic volume were found in 44% of mammography facilities overall, with shortages of radiologists higher in not-for-profit versus for-profit facilities (60% vs 28% reported). Shortages of Mammography Quality Standards Act-qualified technologists were reported by 20% of facilities, with 46% reporting some level of difficulty in maintaining qualified technologists. Waiting times for diagnostic mammography ranged from less than 1 week to 4 weeks, with 85% performed within 1 week. Waiting times for screening mammography ranged from less than 1 week to 8 weeks, with 59% performed between 1 week and 4 weeks. Waiting times for both diagnostic and screening services were two to three times higher in high-volume compared with low-volume facilities. CONCLUSION: Survey results show shortages of radiologists and certified mammography technologists. PMID- 15798154 TI - Pulsed high-intensity focused ultrasound enhances systemic administration of naked DNA in squamous cell carcinoma model: initial experience. AB - PURPOSE: To determine whether exposures to pulsed high-intensity focused ultrasound can enhance local delivery and expression of a reporter gene, administered with systemic injection of naked DNA, in tumors in mice. MATERIALS AND METHODS: The study was performed according to an approved animal protocol and in compliance with guidelines of the institutional animal care and use committee. Squamous cell carcinoma (SCC7) tumors were induced subcutaneously in both flanks of female C3H mice (n = 3) and allowed to grow to average size of 0.4 cm(3). In each mouse, one tumor was exposed to pulsed high-intensity focused ultrasound while a second tumor served as a control. Immediately after ultrasound exposure, a solution containing a cytomegalovirus-green fluorescent protein (GFP) reporter gene construct was injected intravenously via the tail vein. The mouse was sacrificed 24 hours later. Tissue specimens were viewed with fluorescence microscopy to determine the presence of GFP expression, and Western blot analysis was performed, at which signal intensities of expressed GFP were quantitated. A paired Student t test was used to compare mean values in controls with those in treated tumors. Histologic analyses were performed with specific techniques (hematoxylin-eosin staining, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) to determine whether tumor cells had been damaged by ultrasound exposure. RESULTS: GFP expression was present in all sections of tumors that received ultrasound exposure but not in control tumors. Results of signal intensity measurement at Western blot analysis showed expressed GFP to be nine times greater in ultrasound-exposed tumors (160.2 +/- 24.5 [standard deviation]) than in controls (17.4 +/- 11.8) (P = .004, paired Student t test). Comparison of histologic sections from treated tumors with those from controls revealed no destructive effects from ultrasound exposure. CONCLUSION: Local exposure to pulsed high-intensity focused ultrasound in tumors can enhance the delivery and expression of systemically injected naked DNA. PMID- 15798155 TI - Tracheobronchomalacia: comparison between end-expiratory and dynamic expiratory CT for evaluation of central airway collapse. AB - PURPOSE: To compare dynamic expiratory and end-expiratory computed tomography (CT) for depicting central airway collapse in patients with acquired tracheobronchomalacia (TBM). MATERIALS AND METHODS: Institutional review board approval was obtained, and informed consent was not needed. Retrospective review was performed of all patients with a CT diagnosis of TBM in a 10-month period (n = 34) who underwent evaluation of airway disease by means of three different sequences at multi-detector row CT: end inspiration, dynamic expiration, and end expiration (the latter was performed only at the levels of the aortic arch, carina, and bronchus intermedius). Fourteen patients (11 men, three women; age range, 19-79 years) who had comparable images obtained with all three sequences at any of these three levels were included in the study. The degree of airway collapse was measured by two thoracic radiologists in consensus by calculating the percentage change in the area of the airway between inspiratory and expiratory scanning. Statistical analysis was performed by using the paired t test. RESULTS: Dynamic expiratory CT elicited a significantly greater degree of airway collapse than end-expiratory CT at all three levels (P < .005). The mean percentages of airway collapse at each of the three levels were as follows: aortic arch, 53.9% with dynamic expiration versus 35.7% with end expiration (P = .0046); carina, 53.6% with dynamic expiration versus 30.9% with end expiration (P < .0001); and bronchus intermedius, 57.5% with dynamic expiration versus 28.6% with end expiration (P = .0022). CONCLUSION: Dynamic expiratory CT elicits a significantly greater degree of airway collapse than standard end-expiratory CT in patients with TBM. PMID- 15798157 TI - Assessment of apparent diffusion coefficient in normal and degenerated intervertebral lumbar disks: initial experience. AB - PURPOSE: To determine prospectively the diffusibility of water in normal lumbar disks in adults by using the mean apparent diffusion coefficient (ADC) and to determine if a relationship exists between disk ADC and magnetic resonance (MR) findings of disk degeneration. MATERIALS AND METHODS: The study was approved by the Institutional Review Board, and all participants gave written informed consent prior to enrollment. Diffusion-weighted MR imaging of the lumbar spine was performed in 39 patients (all men; mean age, 53 years) and five volunteers (all men; mean age, 31 years). ADC values were recorded for each disk. All disks were visually graded by two independent observers as being normal or as showing at least one of three MR findings of degeneration on sagittal T2-weighted images. Mean ADC values of normal disks were compared with those of degenerated disks and were correlated with age and anatomic location. Data were analyzed by using Kendall correlation statistics, Mantel-Haenszel statistics, and a paired two tailed Student t test. RESULTS: The mean ADC value was 2.27 x 10(-3) mm(2)/sec +/ 0.58 (+/- standard deviation) in normal disks and 2.06 x 10(-3) mm(2)/sec +/- 0.47 in abnormal disks (9% reduction, P = .006). A statistically significant dependence of lumbar disk ADC on anatomic location was reported (analysis of variance, P < .001), with lower ADC values seen in more caudal disks. There was no association between age and mean disk ADC. CONCLUSION: A statistically significant decrease was seen in the ADC values of degenerated lumbar disks when compared with ADC values of normal disks. More caudal disks, even when normal, showed lower ADC values than more cephalic disks. PMID- 15798156 TI - Porcine liver: morphologic characteristics and cell viability at experimental radiofrequency ablation with internally cooled electrodes. AB - PURPOSE: To evaluate morphologic characteristics and cell viability of radiofrequency ablation zones in porcine liver. MATERIALS AND METHODS: Approval of the study protocol was obtained from the Ethics Committee on Use of Live Animals for Teaching and Research at University of Hong Kong. Internally cooled electrodes were used to produce 120 ablated zones ex vivo and 60 ablated zones in vivo with single electrodes (1-, 2-, and 3-cm exposed lengths) or clustered electrodes (1.0-, 2.0-, and 2.5-cm exposed lengths) at 4, 8, 12, and 16 minutes of ablation (ex vivo) and 8 and 12 minutes of ablation (in vivo). Morphologic measurements of each ablated zone were performed. Cell viability in each ablated zone was assessed qualitatively with histochemical staining and quantitatively with measurement of intracellular adenosine 5'-triphosphate (ATP) concentration. RESULTS: Exposed length of electrode (coefficient = 0.79, standard error = 0.04, P < .001), duration of ablation (coefficient = 0.14, standard error = 0.01, P < .001), and clustered electrode design (coefficient = 1.21, standard error = 0.05, P < .001) were independent factors that affected minimal transverse diameter and volume of ablated zone in ex vivo study. Similar morphologic characteristics existed among ablated zones in in vivo study. Mean distance of ablation beyond the electrode tip remained constant (ex vivo, 1.0 cm +/- 0.08 [standard deviation]; in vivo, 0.5 cm +/- 0.05) regardless of different ablation conditions. Histochemical staining revealed no viable hepatocytes from center to margins of white zone in each ablated area. Mean intracellular ATP concentration in margins of white zone (9.5 x 10(-12) mol/microg DNA +/- 1.43) was lower than that in red zone (4088 x 10(-12) mol/microg DNA +/- 65.97, P < .001) and in adjacent normal liver (4528 x 10(-12) mol/microg DNA +/- 52.74, P < .001). CONCLUSION: Distance of ablation beyond the tip of the electrode remained constant (ex vivo, 1.0 cm; in vivo, 0.5 cm) with different conditions of ablation. Complete and uniform cellular destruction was achieved in the white zone of ablated area. PMID- 15798159 TI - Contrast agents for imaging tumor angiogenesis: is bigger better? PMID- 15798158 TI - Hypotensive patients with blunt abdominal trauma: performance of screening US. AB - PURPOSE: To determine retrospectively the accuracy of screening ultrasonography (US) in patients with hypotension (systolic blood pressure or =2) (consensus by three readers) and were assigned to a low- or high-risk group according to the presence of hematuria and/or axial fracture on radiographs. Screening US results were compared with findings with the best available reference standard (computed tomography [CT]), repeat US, other diagnostic test, laparotomy, autopsy, clinical course). Data were compared by using chi(2) or Fisher exact test, depending on expected frequencies, with Bonferroni correction for multiple comparisons. Continuous variables were compared by using unpaired Student t test or Mann-Whitney U test, depending on data distribution. RESULTS: The study included 77 male and 51 female patients (mean age, 42 years). Sensitivity was 85% (44 of 52) for detection of any injuries, 97% (30 of 31) for surgical injuries (ie, injuries requiring surgery), and 100% (10 of 10) for fatal injuries. Specificity was 96% (73 of 76), 82% (80 of 97), and 69% (81 of 118), and accuracy was 91% (117 of 128), 86% (110 of 128), and 71% (91 of 128), for respective injury categories. One nonfatal surgical injury was missed in a high-risk patient. For each injury category, frequency of injury in patients with a fluid score of 2 or more was nine times that in patients with a score of 0 (P < .001 for all comparisons). Frequency of false-negative US findings in high-risk patients was eight times that in low-risk patients (P < .01). CONCLUSION: In patients who are hypotensive after blunt abdominal trauma and not hemodynamically stable enough to undergo diagnostic CT, negative US findings virtually exclude surgical injury, while positive US findings indicate surgical injury in 64% of cases. PMID- 15798161 TI - Is infiltrate a useful term in the interpretation of chest radiographs? Physician survey results. AB - PURPOSE: To determine how physicians interpret the word infiltrate when it is used in a chest radiography report and if the word is helpful in the clinical management of patients. MATERIALS AND METHODS: Informed consent from participants was not required, and the study protocol was granted exempt status. One hundred sixty-five questionnaires were distributed, in conjunction with various physician meetings, at three separate nonaffiliated training hospitals. In the three question survey, respondents were asked the definition of infiltrate, whether the term is helpful in guiding patient therapy, and whether it implies an etiology. The respondent demographic data obtained included date of graduation from medical school, amount of postgraduate training completed, primary or training specialty, and board or sub-board qualification. RESULTS: There were 151 physician respondents, 94 (62.3%) of whom were house staff members. One hundred fifteen (76%) responders chose bacterial pneumonia as a condition consistent with infiltrate. One hundred thirty (86.1%) respondents replied that infiltrate implied more than one pathophysiologic condition. Eighty-two (54.3%) of those surveyed thought infiltrate could mean any of six or more different pathophysiologic conditions, including nonspecific pneumonia, interstitial pneumonia, viral pneumonia, consolidation, or nonspecific interstitial process. The number of terms selected did not vary according to level of residency training (P = .23); however, there was a significant specialty-related difference in the number of terms selected (P = .018). Internists selected a median of 10 terms, while others selected a median of six. Only 54 (36%) respondents thought that the term infiltrate was helpful in patient care, and only five (3%) thought that the term implied an etiology. CONCLUSION: Infiltrate is a nonspecific and imprecise term when it is used as a radiograph descriptor, and use of this term does not usually enhance patient care. PMID- 15798163 TI - Colonoscopy: virtual and optical--another look, another view. PMID- 15798162 TI - Prevalence and costs of chronic disease in a health care system structured for treatment of acute illness. PMID- 15798164 TI - Building a CT colonography program: necessary ingredients for reimbursement and clinical success. PMID- 15798165 TI - Comparison of recall and cancer detection rates for immediate versus batch interpretation of screening mammograms. AB - PURPOSE: To retrospectively compare recall and cancer detection rates between immediate and subsequent batch methods for interpretation of screening mammograms. MATERIALS AND METHODS: Institutional review board approval was obtained, and informed consent was waived. Retrospective analysis was performed for 8698 screening mammograms obtained between January 1 and October 31, 2001, which were interpreted either immediately (n = 4113) or subsequently with batch method (n = 4585). Data were collected from data reporting system and patient billing records. Patients with high risk factors were excluded; 3441 patients were in the immediate group, and 3932 were in the batch group. The two groups were compared with respect to age, breast density, and availability of comparison films with Wilcoxon rank sum test. Recall rates and cancer detection rates for each group were determined and compared with Pearson chi(2) test; false-negative rates were compared with Fischer exact test. RESULTS: A significant difference (P < .001) was noted in recall rates between immediate (18%) and batch (14%) groups; however, no significant difference (P = .7) was noted in cancer detection rates (immediate, 0.5%; batch, 0.4%). Mean age of patients was 56.8 years (age range, 21-96 years) in the immediate group and 56.2 years (age range 24-98 years) in the batch group (P = .02). Comparison of breast densities between groups indicates no statistically significant difference (P = .4). The batch group had significantly fewer comparison mammograms (3106 [79%]) available than the immediate group (2856 [83%]) (P < .001). There was no significant difference in false-negative rates between the immediate group (0.1%) and the batch group (0.1%) (P > .99). CONCLUSION: Immediate interpretation of screening mammograms resulted in a statistically significant increase in recalls and additional clinical work-ups of perceived abnormalities; however, no significant difference in cancer detection rate was detected between groups. PMID- 15798168 TI - Case 81: antiphospholipid antibody syndrome with adrenal hemorrhage and Budd Chiari syndrome. PMID- 15798166 TI - Hormone replacement therapy in postmenopausal women: breast tissue perfusion determined with MR imaging--initial observations. AB - PURPOSE: To use magnetic resonance (MR) imaging to evaluate tissue perfusion in the normal breast parenchyma of postmenopausal women with current or recent hormone replacement therapy (HRT). MATERIALS AND METHODS: The study was approved by the institutional subcommittee on human studies, and informed consent was obtained from all patients prior to MR imaging. Sixty postmenopausal women (age range, 44-77 years) were grouped according to HRT received: estrogen replacement therapy (ERT) (n = 13), combined (estrogen and progesterone) replacement therapy (CRT) (n = 16), selective estrogen receptor modulator (SERM) therapy (n = 8), and no (hormone replacement) therapy (NT) (n = 23). MR imaging with a 1.5-T magnet was performed by using gradient-echo and dynamic contrast material-enhanced echo planar pulse sequences before and after gadopentetate dimeglumine injection. Precontrast T1 relaxation times were measured, after which extraction-flow product (EFP) maps were calculated with a multicompartmental model. Analysis of variance was performed. RESULTS: Age did not significantly differ between the groups (P > .3). Women receiving ERT or CRT at the time of MR imaging had higher EFP values (7.3 mL . 100 g(-1) . min(-1)+/- 2.6 and 7.1 mL . 100 g(-1) . min( 1)+/- 3.8, respectively) than did women receiving NT (4.4 mL . 100 g(-1) . min( 1)+/- 2.1) (P = .012 and P = .008, respectively) or SERM therapy (3.9 mL . 100 g( 1) . min(-1)+/- 1.1) (P = .015 and P = .013, respectively). Women who ended ERT or CRT 1-47 months before MR examination had lower EFP values than did women with current ERT or CRT and had higher EFP values than did women receiving NT or SERM therapy (6.2 mL . 100 g(-1) . min(-1)+/- 2.4 and 5.9 mL . 100 g(-1) . min(-1)+/- 3.8, respectively), but the observed differences were not significant (P > .1). Differences in T1 between all groups were not significant (P > .5). CONCLUSION: Higher breast tissue perfusion is observed in postmenopausal women receiving HRT. PMID- 15798169 TI - Relative procedure intensity with self-referral and radiologist referral: extremity radiography. AB - PURPOSE: To compare the relative use of bilateral versus unilateral extremity radiographic examinations when patients are referred to radiologists for imaging (radiologist referred) versus when studies are performed in the referring physician's office (self-referred). MATERIALS AND METHODS: We reviewed 1 year of claims data for extremity radiographic examinations performed by a referring physician or referred to a radiology facility and claims data for related patient office visits. Data were analyzed for orthopedics, podiatry, and rheumatology, and data were divided by the practice pattern of the referring physician into pure self-referring, pure radiologist-referring, and mixed-referring categories. We compared the percentage of unilateral and bilateral studies and the number of unilateral and bilateral studies per 100 office visits in each setting. Statistical analysis of each comparison was performed with a one-tailed Z test. RESULTS: A total of 13 094 (14%) self-referred studies were bilateral, while 778 (10%) radiologist-referred studies were bilateral (P < .001). The rate of self referred bilateral examinations was 2.21 times higher per 100 office visits than the rate of radiologist-referred bilateral examinations. Combined bilateral and unilateral use by self-referrers was only 1.86 times higher than use by radiologist-referrers. Orthopedists had no clinically meaningful difference in the percentage of self-referred and radiologist-referred bilateral studies, but they ordered 1.98 times as many studies per 100 visits when they self-referred studies. Self-referring podiatrists and rheumatologists ordered bilateral studies up to 3.25 times more frequently than did their radiologist-referring colleagues. Mixed-referring podiatrists had 2.70-times increased use of bilateral examinations when performing imaging in their offices, whereas mixed-referring rheumatologists had 6.40-times increase in that setting. CONCLUSION: Orthopedists, podiatrists, and rheumatologists use extremity radiography at a higher rate when they self-refer. Moreover, self-referring podiatrists and rheumatologists order radiographic examinations of increased intensity compared with radiologist-referring physicians. PMID- 15798170 TI - Detection of single and multiple targets in tissue phantoms with fluorescence enhanced optical imaging: feasibility study. AB - PURPOSE: To evaluate the feasibility of reconstructing single and multiple targets by using fluorescence-enhanced tomography of a breast tissue phantom. MATERIALS AND METHODS: Frequency-domain fluorescence measurements were performed in cup-shaped tissue phantoms of clinically relevant size (diameter, 10 cm) in which single or multiple 0.5-1.0-cm(3) targets that contained micromolar concentrations of indocyanine green with 1:0 and 100:1 target-to-background (T:B) contrast ratios had been embedded. Rapid acquisition of time-dependent fluorescent light measurements was performed at the phantom surface in response to point illumination of excitation light by using a gain-modulated intensified charge-coupled device detection system. Boundary surface measurements were used to tomographically reconstruct the interior targets located with various experimental conditions. RESULTS: Single 1.0-cm(3) targets located between 1.43 and 2.82 cm deep from the phantom surface at a T:B contrast ratio of 100:1 and three approximately 0.55-cm(3) targets located about 1.30 cm deep at a T:B contrast ratio of 1:0 were reconstructed with minimal or no artifacts by using boundary surface fluorescence measurements and an approximate extended Kalman filter algorithm. CONCLUSION: It is feasible to detect single or multiple fluorescent targets in tissue phantoms of clinically relevant size by using fluorescence-enhanced optical tomography. PMID- 15798171 TI - The hot nose sign. PMID- 15798172 TI - Comparison of hybrid echo-planar imaging and FLASH myocardial perfusion cardiovascular MR imaging. AB - The purpose of this study was to compare fast single-shot gradient-echo (FLASH) and hybrid echo-planar imaging (EPI) magnetic resonance (MR) technologies regarding the relative contrast-to-noise ratio (CNR), spatiotemporal resolution, size of inducible perfusion defects, and presence of artifacts in patients with coronary artery disease (CAD). Fifteen patients with CAD underwent rest and adenosine stress gadolinium first-pass perfusion cardiovascular MR examinations with EPI and FLASH. The study was approved by the local ethics committee, and each subject gave written informed consent. The spatial resolution of the two sequences was made similar in nine patients, and the temporal resolution was made similar in six. The images were assessed for CNR, artifact, and size of inducible perfusion defects. The CNR was significantly higher with the EPI sequence, whether matched for spatial (32 vs 22 [46%], P < .001) or temporal (35 vs 23 [51%], P < .001) resolution. There was no significant difference in scoring for artifact or area and transmural extent of inducible perfusion defects with EPI and FLASH, whether matched for temporal or spatial resolution. Further work is warranted to determine the relative diagnostic accuracy of the two techniques. PMID- 15798173 TI - Thoracic masses treated with percutaneous cryotherapy: initial experience with more than 200 procedures. AB - PURPOSE: To perform and report initial experience with percutaneous cryotherapy (PCT) of the thorax. MATERIALS AND METHODS: A human investigation committee approved the study protocol, and all patients gave informed consent. One hundred eighty-seven patients who were not surgical candidates underwent computed tomography (CT)-guided PCT for treatment of thoracic cancer masses. CT-visualized low-attenuating ice formation after PCT was compared with initial tumor size and location. At 1 week and at 1, 3, 6, and 12 months after PCT, the various findings seen on available CT scans and any complications were noted. chi(2) and Student t tests were used to identify significant differences in frequencies and mean values of imaging observations, respectively. RESULTS: Ice formation was identified at CT as reduced attenuation values (in Hounsfield units) within soft tissue masses, the mean sizes of which were 4.3 cm +/- 0.2 (standard deviation) in peripheral locations and 6.4 cm +/- 0.3 in central locations. Tumor size and location were independent predictors of tumor coverage by low-attenuating ice: Mean coverage was 99% for peripheral masses 4 cm or smaller (n = 101) and 80% for central masses larger than 4 cm (n = 58) (P < .001). An area of necrotic cavitation larger than the original mass developed in 80% (77 of 96) of masses within 1 week and was nearly resolved by 3 months in 7% (five of 76) of masses. By 6 months, minimal pulmonary scarring was noted in 56 patients and 86% of masses showed reduced or stable size. The overall rate of pneumothorax was only 12% (22 of 187 patients), and other side effects appeared to be self limited. No major bleeding or bronchial damage was noted. Two deaths in debilitated patients were temporally related, and two complications involved brachial and recurrent laryngeal nerve damage. The patient with laryngeal nerve damage regained speech within 2 months. CONCLUSION: CT-guided PCT yielded low procedural morbidity given the extent of freezing, even near mediastinal structures. Ongoing advances in cryotechnology, imaging guidance, and treatment planning may help to avoid the degree of undertreatment of larger central masses observed in this study. PMID- 15798174 TI - Automatic exposure control techniques for individual dose adaptation. PMID- 15798175 TI - Digital radiography of the chest: reassessment of the high-voltage technique? PMID- 15798176 TI - Analysis of atherosclerosis with micro-CT. PMID- 15798177 TI - Hormone receptor status in patients with breast cancer and liver metastases treated with laser-induced intersititial thermotherapy. PMID- 15798179 TI - Human progesterone receptor displays cell cycle-dependent changes in transcriptional activity. AB - The human progesterone receptor (PR) contains multiple Ser-Pro phosphorylation sites that are potential substrates for cyclin-dependent kinases, suggesting that PR activity might be regulated during the cell cycle. Using T47D breast cancer cells stably transfected with an mouse mammary tumor virus (MMTV) chloramphenicol acetyltransferase reporter (Cat0) synchronized in different phases of the cell cycle, we found that PR function and phosphorylation is remarkably cell cycle dependent, with the highest activity in S phase. Although PR expression was reduced in the G2/M phase, the activity per molecule of receptor was markedly reduced in both G1 and G2/M phases compared to the results seen with the S phase of the cell cycle. Although PR is recruited to the MMTV promoter equivalently in the G1 and S phases, recruitment of SRC-1, SRC-3, and, consequently, CBP is reduced in G1 phase despite comparable expression levels of SRC-1 and SRC-3. In G2/M phase, site-specific phosphorylation of PR at Ser162 and at Ser294, a site previously reported to be critical for transcriptional activity and receptor turnover, was abolished. Treatment with the histone deacetylase inhibitor trichostatin A elevated G1 and G2/M activity to that of the S phase, indicating that the failure to recruit sufficient levels of active histone acetyltransferase is the primary defect in PR-mediated transactivation. PMID- 15798180 TI - Numb proteins specify asymmetric cell fates via an endocytosis- and proteasome independent pathway. AB - Numb proteins are evolutionarily conserved signaling molecules that make the daughter cells different after asymmetric divisions by segregating to only one daughter. They contain distinct binding motifs for alpha-adaptin (alpha-Ada) and proteins with Eps15 homology (EH) domains, which regulate endocytosis, and for E3 ubiquitin ligases, which target proteins for proteasome-mediated degradation. In Drosophila melanogaster, Numb acts by inhibiting Notch activity to cause a bias in Notch-mediated cell-cell communication. These findings have led to the hypothesis that Numb modulates Notch signaling by using endocytosis and proteasomes to directly reduce Notch protein levels at the cell surface. Here we show that two Drosophila EH proteins, Eps15 homologue 1 (EH1) and the dynamin associated 160-kDa protein (Dap160), negatively regulate Notch signaling. However, neither elimination of the binding motifs for endocytic proteins nor simultaneous reduction of proteasome activity affects the activity of Numb proteins. Our findings indicate that an endocytosis- and proteasome-independent pathway may mediate Numb signaling in asymmetric cell fate specification. PMID- 15798178 TI - Class II histone deacetylases: from sequence to function, regulation, and clinical implication. PMID- 15798181 TI - The phosphoinositide phosphatase Sjl2 is recruited to cortical actin patches in the control of vesicle formation and fission during endocytosis. AB - The Saccharomyces cerevisiae synaptojanin-like proteins (Sjl1, Sjl2, and Sjl3) are phosphoinositide (PI) phosphatases that regulate PI metabolism in the control of actin organization and membrane trafficking. However, the primary sites of action for each of the yeast synaptojanin-like proteins remain unclear. In this study, we show that Sjl2 is localized to cortical actin patches, sites of endocytosis. Cortical recruitment of Sjl2 requires the actin patch component Abp1. Consistent with this, the SH3 domain-containing protein Abp1 physically associates with Sjl2 through its proline-rich domain. Furthermore, abp1Delta mutations confer defects resembling loss of SJL2; sjl1Delta abp1Delta double mutant cells exhibit invaginated plasma membranes and impaired endocytosis, findings similar to those for sjl1Delta sjl2Delta mutant cells. Thus, Abp1 acts as an adaptor protein in the localization or concentration of Sjl2 during late stages of endocytic vesicle formation. Overexpression of the Hip1-related protein Sla2 delayed the formation of extended plasma membrane invaginations in sjl2ts cells, indicating that Sla2 may become limiting or misregulated in cells with impaired PI phosphatase activity. Consistent with this, the cortical actin patch protein Sla2 is mislocalized in sjl1Delta sjl2Delta mutant cells. Together, our studies suggest that PI metabolism by the synaptojanin-like proteins coordinately directs actin dynamics and membrane invagination, in part by regulation of Sla2. PMID- 15798183 TI - Identification of cis-regulatory sequences in the human angiotensinogen gene by transgene coplacement and site-specific recombination. AB - The function of putative regulatory sequences identified in cell transfection experiments can be elucidated only through in vivo experimentation. However, studies of gene regulation in transgenic mice (TgM) are often compromised by the position effects, in which independent transgene insertions differ in expression depending on their location in the genome. In order to overcome such a dilemma, a method called transgene coplacement has been developed in Drosophila melanogaster. In this method, any two sequences can be positioned at exactly the same genomic site by making use of Cre/loxP recombination. Here we applied this method to mouse genetics to characterize the function of direct repeat (DR) sequences in the promoter of the human angiotensinogen (hAGT) gene, the precursor of the vasoactive octapeptide angiotensin II. We modified a hAGT bacterial artificial chromosome to use Cre/loxP recombination in utero to generate TgM lines bearing a wild-type or a mutant promoter-driven hAGT locus integrated at a single chromosomal position. The expression analyses revealed that DR sequences contribute 50 or >95% to hAGT transcription in the liver and kidneys, respectively, whereas same sequences are not required in the heart and brain. This is the first in vivo dissection of DNA cis elements that are demonstrably indispensable for regulating both the level and cell type specificity of hAGT gene transcription. PMID- 15798182 TI - MRG15 regulates embryonic development and cell proliferation. AB - MRG15 is a highly conserved protein, and orthologs exist in organisms from yeast to humans. MRG15 associates with at least two nucleoprotein complexes that include histone acetyltransferases and/or histone deacetylases, suggesting it is involved in chromatin remodeling. To study the role of MRG15 in vivo, we generated knockout mice and determined that the phenotype is embryonic lethal, with embryos and the few stillborn pups exhibiting developmental delay. Immunohistochemical analysis indicates that apoptosis in Mrg15-/- embryos is not increased compared with wild-type littermates. However, the number of proliferating cells is significantly reduced in various tissues of the smaller null embryos compared with control littermates. Cell proliferation defects are also observed in Mrg15-/- mouse embryonic fibroblasts. The hearts of the Mrg15-/- embryos exhibit some features of hypertrophic cardiomyopathy. The increase in size of the cardiomyocytes is most likely a response to decreased growth of the cells. Mrg15-/- embryos appeared pale, and microarray analysis revealed that alpha-globin gene expression was decreased in null versus wild-type embryos. We determined by chromatin immunoprecipitation that MRG15 was recruited to the alpha globin promoter during dimethyl sulfoxide-induced mouse erythroleukemia cell differentiation. These findings demonstrate that MRG15 has an essential role in embryonic development via chromatin remodeling and transcriptional regulation. PMID- 15798185 TI - 17beta-estradiol inhibits inflammatory gene expression by controlling NF-kappaB intracellular localization. AB - Estrogen is an immunoregulatory agent, in that hormone deprivation increases while 17beta-estradiol (E2) administration blocks the inflammatory response; however, the underlying mechanism is still unknown. The transcription factor p65/relA, a member of the nuclear factor kappaB (NF-kappaB) family, plays a major role in inflammation and drives the expression of proinflammatory mediators. Here we report a novel mechanism of action of E2 in inflammation. We observe that in macrophages E2 blocks lipopolysaccharide-induced DNA binding and transcriptional activity of p65 by preventing its nuclear translocation. This effect is selectively activated in macrophages to prevent p65 activation by inflammatory agents and extends to other members of the NF-kappaB family, including c-Rel and p50. We observe that E2 activates a rapid and persistent response that involves the activation of phosphatidylinositol 3-kinase, without requiring de novo protein synthesis or modifying Ikappa-Balpha degradation and mitogen-activated protein kinase activation. Using a time course experiment and the microtubule disrupting agent nocodazole, we observe that the hormone inhibits p65 intracellular transport to the nucleus. This activity is selectively mediated by estrogen receptor alpha (ERalpha) and not ERbeta and is not shared by conventional anti-inflammatory drugs. These results unravel a novel and unique mechanism for E2 anti-inflammatory activity, which may be useful for identifying more selective ligands for the prevention of the inflammatory response. PMID- 15798184 TI - Spatial distribution and function of sterol regulatory element-binding protein 1a and 2 homo- and heterodimers by in vivo two-photon imaging and spectroscopy fluorescence resonance energy transfer. AB - Sterol regulatory element-binding proteins (SREBPs) are a subfamily of basic helix-loop-helix-leucine zipper proteins that regulate lipid metabolism. We show novel evidence of the in vivo occurrence and subnuclear spatial localization of both exogenously expressed SREBP-1a and -2 homodimers and heterodimers obtained by two-photon imaging and spectroscopy fluorescence resonance energy transfer. SREBP-1a homodimers localize diffusely in the nucleus, whereas SREBP-2 homodimers and the SREBP-1a/SREBP-2 heterodimer localize predominantly to nuclear speckles or foci, with some cells showing a diffuse pattern. We also used tethered SREBP dimers to demonstrate that both homo- and heterodimeric SREBPs activate transcription in vivo. Ultrastructural analysis revealed that the punctate foci containing SREBP-2 are electron-dense nuclear bodies, similar or identical to structures containing the promyelocyte (PML) protein. Immunofluorescence studies suggest that a dynamic interplay exists between PML, as well as another component of the PML-containing nuclear body, SUMO-1, and SREBP-2 within these nuclear structures. These findings provide new insight into the overall process of transcriptional activation mediated by the SREBP family. PMID- 15798187 TI - Genome-wide prediction and analysis of yeast RNase III-dependent snoRNA processing signals. AB - In Saccharomyces cerevisiae, the maturation of both pre-rRNA and pre-small nucleolar RNAs (pre-snoRNAs) involves common factors, thereby providing a potential mechanism for the coregulation of snoRNA and rRNA synthesis. In this study, we examined the global impact of the double-stranded-RNA-specific RNase Rnt1p, which is required for pre-rRNA processing, on the maturation of all known snoRNAs. In silico searches for Rnt1p cleavage signals, and genome-wide analysis of the Rnt1p-dependent expression profile, identified seven new Rnt1p substrates. Interestingly, two of the newly identified Rnt1p-dependent snoRNAs, snR39 and snR59, are located in the introns of the ribosomal protein genes RPL7A and RPL7B. In vitro and in vivo experiments indicated that snR39 is normally processed from the lariat of RPL7A, suggesting that the expressions of RPL7A and snR39 are linked. In contrast, snR59 is produced by a direct cleavage of the RPL7B pre mRNA, indicating that a single pre-mRNA transcript cannot be spliced to produce a mature RPL7B mRNA and processed by Rnt1p to produce a mature snR59 simultaneously. The results presented here reveal a new role of yeast RNase III in the processing of intron-encoded snoRNAs that permits independent regulation of the host mRNA and its associated snoRNA. PMID- 15798186 TI - A novel SR-related protein is required for the second step of Pre-mRNA splicing. AB - The SR family proteins and SR-related polypeptides are important regulators of pre-mRNA splicing. A novel SR-related protein of an apparent molecular mass of 53 kDa was isolated in a gene trap screen that identifies proteins which localize to the nuclear speckles. This novel protein possesses an arginine- and serine-rich domain and was termed SRrp53 (for SR-related protein of 53 kDa). In support for a role of this novel RS-containing protein in pre-mRNA splicing, we identified the mouse ortholog of the Saccharomyces cerevisiae U1 snRNP-specific protein Luc7p and the U2AF65-related factor HCC1 as interacting proteins. In addition, SRrp53 is able to interact with some members of the SR family of proteins and with U2AF35 in a yeast two-hybrid system and in cell extracts. We show that in HeLa nuclear extracts immunodepleted of SRrp53, the second step of pre-mRNA splicing is blocked, and recombinant SRrp53 is able to restore splicing activity. SRrp53 also regulates alternative splicing in a concentration-dependent manner. Taken together, these results suggest that SRrp53 is a novel SR-related protein that has a role both in constitutive and in alternative splicing. PMID- 15798188 TI - Scaffold attachment factor B1 functions in development, growth, and reproduction. AB - Scaffold attachment factor B1 (SAFB1) is a multifunctional protein that can bind both DNA and RNA and is involved in RNA processing and stress response. In addition, SAFB1 contains a transcriptional repression domain and can bind certain hormone receptors and repress their activity. To assess the role of SAFB1 in vivo, we generated SAFB1 mutant mice through targeted deletion in embryonic stem cells. While viable homozygous mutant (SAFB1-/-) mice were obtained, genotypic distribution indicated that homozygous deficiency resulted in both prenatal and neonatal lethality. Mice lacking SAFB1 exhibited dwarfism, as a result of in utero growth retardation, and had low serum insulin-like growth factor 1 (IGF1) levels. In agreement with the previous characterization of SAFB1 as a corepressor for hormone receptors, we found that SAFB1-/- mice displayed dramatic defects in the development and function of the reproductive system. Male SAFB1 null mice were infertile, apparently because of low circulating levels of testosterone. SAFB1-/- testes were small and showed progressive degeneration of the germinal epithelium, increased apoptosis of germ cells, and Leydig cell hyperplasia. SAFB /- female mice were subfertile and showed progressive infertility, in part because of defects in oviductal transport and reduced numbers of follicles. Immortalized SAFB1-/- mouse embryonic fibroblasts showed cell-intrinsic defects including increased transcriptional estrogen receptor alpha activity and enhanced responsiveness to IGF1. Together, these in vivo findings establish a critical role for SAFB1 in development, growth regulation, and reproduction. PMID- 15798189 TI - Identification of a novel, intraperoxisomal pex14-binding site in pex13: association of pex13 with the docking complex is essential for peroxisomal matrix protein import. AB - The peroxisomal docking complex is a key component of the import machinery for matrix proteins. The core protein of this complex, Pex14, is thought to represent the initial docking site for the import receptors Pex5 and Pex7. Associated with this complex is a fraction of Pex13, another essential component of the import machinery. Here we demonstrate that Pex13 directly binds Pex14 not only via its SH3 domain but also via a novel intraperoxisomal site. Furthermore, we demonstrate that Pex5 also contributes to the association of Pex13 with Pex14. Peroxisome function was affected only mildly by mutations within the novel Pex14 interaction site of Pex13 or by the non-Pex13-interacting mutant Pex5(W204A). However, when these constructs were tested in combination, PTS1-dependent import and growth on oleic acid were severely compromised. When the SH3 domain-mediated interaction of Pex13 with Pex14 was blocked on top of that, PTS2-dependent matrix protein import was completely compromised and Pex13 was no longer copurified with the docking complex. We conclude that the association of Pex13 with Pex14 is an essential step in peroxisomal protein import that is enabled by two direct interactions and by one that is mediated by Pex5, a result which indicates a novel, receptor-independent function of Pex5. PMID- 15798190 TI - Imprinted Nesp55 influences behavioral reactivity to novel environments. AB - Genomic imprinting results in parent-of-origin-dependent monoallelic expression of selected genes. Although their importance in development and physiology is recognized, few imprinted genes have been investigated for their effects on brain function. Gnas is a complex imprinted locus whose gene products are involved in early postnatal adaptations and neuroendocrine functions. Gnas encodes the stimulatory G-protein subunit Gsalpha and two other imprinted protein-coding transcripts. Of these, the Nesp transcript, expressed exclusively from the maternal allele, codes for neuroendocrine secretory protein 55 (Nesp55), a chromogranin-like polypeptide associated with the constitutive secretory pathway but with an unknown function. Nesp is expressed in restricted brain nuclei, suggesting an involvement in specific behaviors. We have generated a knockout of Nesp55 in mice. Nesp55-deficient mice develop normally, excluding a role of this protein in the severe postnatal effects associated with imprinting of the Gnas cluster. Behavioral analysis of adult Nesp55 mutants revealed, in three separate tasks, abnormal reactivity to novel environments independent of general locomotor activity and anxiety. This phenotype may be related to prominent Nesp55 expression in the noradrenergic locus coeruleus. These results indicate a role of maternally expressed Nesp55 in controlling exploratory behavior and are the first demonstration that imprinted genes affect such a fundamental behavior. PMID- 15798191 TI - dDP is needed for normal cell proliferation. AB - To gain insight into the essential functions of E2F, we have examined the phenotypes caused by complete inactivation of E2F and DP family members in Drosophila. Our results show that dDP requires dE2F1 and dE2F2 for DNA-binding activity in vitro and in vivo. In tissue culture cells and in mutant animals, the levels of dE2F and dDP proteins are strongly interdependent. In the absence of dDP, the levels of dE2F1 and dE2F2 decline dramatically, and vice versa. Accordingly, the cell cycle and transcriptional phenotypes caused by targeting dDP mimic the effects of targeting both dE2F1 and dE2F2 and are indistinguishable from the effects of inactivating all three proteins. Although trans-heterozygous dDP mutant animals develop to late pupal stages, the analysis of somatic mutant clones shows that dDP mutant cells are at a severe proliferative disadvantage when compared directly with wild-type neighbors. Strikingly, the timing of S phase entry or exit is not delayed in dDP mutant clones, nor is the accumulation of cyclin A or cyclin B. However, the maximal level of bromodeoxyuridine incorporation is reduced in dDP mutant clones, and RNA interference experiments show that dDP-depleted cells are prone to stall in S phase. In addition, dDP mutant clones contain reduced numbers of mitotic cells, indicating that dDP mutant cells have a defect in G2/M-phase progression. Thus, dDP is not essential for developmental control of the G1-to-S transition, but it is required for normal cell proliferation, for optimal DNA synthesis, and for efficient G2/M progression. PMID- 15798192 TI - Adaptive myogenesis under hypoxia. AB - Previous studies have indicated that myoblasts can differentiate and repair muscle injury after an ischemic insult. However, it is unclear how hypoxia or glucose deprivation in the ischemic microenvironment affects myoblast differentiation. We have found that myogenesis can adapt to hypoxic conditions. This adaptive mechanism is accompanied by initial inhibition of the myoD, E2A, and myogenin genes followed by resumption of their expression in an oxygen dependent manner. The regulation of myoD transcription by hypoxia is correlated with transient deacetylation of histones associated with the myoD promoter. It is noteworthy that, unlike the differentiation of other cell types such as preadipocytes or chondroblasts, the effect of hypoxia on myogenesis is independent of HIF-1, a ubiquitous regulator of transcription under hypoxia. While myogenesis can also adapt to glucose deprivation, the combination of severe hypoxia and glucose deprivation found in an ischemic environment results in pronounced loss of myoblasts. Our studies indicate that the ischemic muscle can be repaired via the adaptive differentiation of myogenic precursors, which depends on the levels of oxygen and glucose in the ischemic microenvironment. PMID- 15798193 TI - PINCH1 plays an essential role in early murine embryonic development but is dispensable in ventricular cardiomyocytes. AB - PINCH1, an adaptor protein composed of five LIM domains, mediates protein-protein interactions and functions as a component of the integrin-integrin-linked kinase (ILK) complex. The integrin-ILK signaling complex plays a pivotal role in cell motility, proliferation, and survival during embryonic development of many animal species. To elucidate the physiological function of PINCH1 in mouse embryonic development, we have deleted the mouse PINCH1 gene by homologous recombination. Mice heterozygous for PINCH1 are viable and indistinguishable from wild-type littermates. However, no viable homozygous offspring were observed from PINCH1+/- intercrosses. Histological analysis of homozygous mutant embryos revealed that they had a disorganized egg cylinder by E5.5, which degenerated by E6.5. Furthermore, E5.5 PINCH1-/- embryos exhibited decreased cell proliferation and excessive cell death. We have also generated and analyzed mice in which PINCH1 has been specifically deleted in ventricular cardiomyocytes. These mice exhibit no basal phenotype, with respect to mouse survival, cardiac histology, or cardiac function as measured by echocardiography. Altogether, these data indicate that PINCH1 plays an essential role in early murine embryonic development but is dispensable in ventricular cardiomyocytes. PMID- 15798194 TI - PKR and GCN2 kinases and guanine nucleotide exchange factor eukaryotic translation initiation factor 2B (eIF2B) recognize overlapping surfaces on eIF2alpha. AB - Four stress-responsive protein kinases, including GCN2 and PKR, phosphorylate eukaryotic translation initiation factor 2alpha (eIF2alpha) on Ser51 to regulate general and gene-specific protein synthesis. Phosphorylated eIF2 is an inhibitor of its guanine nucleotide exchange factor, eIF2B. Mutations that block translational regulation were isolated throughout the N-terminal OB-fold domain in Saccharomyces cerevisiae eIF2alpha, including those at residues flanking Ser51 and around 20 A away in the conserved motif K79GYID83. Any mutation at Glu49 or Asp83 blocked translational regulation; however, only a subset of these mutations impaired Ser51 phosphorylation. Substitution of Ala for Asp83 eliminated phosphorylation by GCN2 and PKR both in vivo and in vitro, establishing the critical contributions of remote residues to kinase-substrate recognition. In contrast, mutations that blocked translational regulation but not Ser51 phosphorylation impaired the binding of eIF2B to phosphorylated eIF2alpha. Thus, two structurally distinct effectors of eIF2 function, eIF2alpha kinases and eIF2B, have evolved to recognize the same surface and overlapping determinants on eIF2alpha. PMID- 15798195 TI - Identification of neuronal enhancers of the proopiomelanocortin gene by transgenic mouse analysis and phylogenetic footprinting. AB - The proopiomelanocortin (POMC) gene is expressed in the pituitary and arcuate neurons of the hypothalamus. POMC arcuate neurons play a central role in the control of energy homeostasis, and rare loss-of-function mutations in POMC cause obesity. Moreover, POMC is the prime candidate gene within a highly significant quantitative trait locus on chromosome 2 associated with obesity traits in several human populations. Here, we identify two phylogenetically conserved neuronal POMC enhancers designated nPE1 (600 bp) and nPE2 (150 bp) located approximately 10 to 12 kb upstream of mammalian POMC transcriptional units. We show that mouse or human genomic regions containing these enhancers are able to direct reporter gene expression to POMC hypothalamic neurons, but not the pituitary of transgenic mice. Conversely, deletion of nPE1 and nPE2 in the context of the entire transcriptional unit of POMC abolishes transgene expression in the hypothalamus without affecting pituitary expression. Our results indicate that the nPEs are necessary and sufficient for hypothalamic POMC expression and that POMC expression in the brain and pituitary is controlled by independent sets of enhancers. Our study advances the understanding of the molecular nature of hypothalamic POMC neurons and will be useful to determine whether polymorphisms in POMC regulatory regions play a role in the predisposition to obesity. PMID- 15798196 TI - Identification of nuclear import and export signals within Fli-1: roles of the nuclear import signals in Fli-1-dependent activation of megakaryocyte-specific promoters. AB - The Ets factor Friend leukemia integration 1 (Fli-1) is an important regulator of megakaryocytic (Mk) differentiation. Here, we demonstrate two novel nuclear localization signals (NLSs) within Fli-1: one (NLS1) is located at the N terminus, and another (NLS2) is within the Ets domain. Nuclear accumulation of Fli-1 reflected the combined functional effects of the two discrete NLSs. Each NLS can independently direct nuclear transport of a carrier protein, with mutations within the NLSs affecting nuclear accumulation. NLS1 has a bipartite motif, whereas the NLS2 region contains a nonclassical NLS. Both NLSs bind importin alpha (IMPalpha) and IMPbeta, with NLS1 and NLS2 being predominantly recognized by IMPalpha and IMPbeta, respectively. Fli-1 also contains one nuclear export signal. Leptomycin B abolished its cytoplasmic accumulation, showing CRM1 dependency. We demonstrate that Ets domain binding to specific target DNA effectively blocks IMP binding, indicating that the targeted DNA binding plays a role in localizing Fli-1 to its destination and releasing IMPs for recycling back to the cytoplasm. Finally, by analyzing full-length Fli-1 carrying NLS1, NLS2, and combined NLS1-NLS2 mutations, we conclude that two functional NLSs exist in Fli-1 and that each NLS is sufficient to target Fli-1 to the nucleus for activation of Mk-specific genes. PMID- 15798197 TI - Coupling of human circadian and cell cycles by the timeless protein. AB - The Timeless protein is essential for circadian rhythm in Drosophila. The Timeless orthologue in mice is essential for viability and appears to be required for the maintenance of a robust circadian rhythm as well. We have found that the human Timeless protein interacts with both the circadian clock protein cryptochrome 2 and with the cell cycle checkpoint proteins Chk1 and the ATR-ATRIP complex and plays an important role in the DNA damage checkpoint response. Down regulation of Timeless in human cells seriously compromises replication and intra S checkpoints, indicating an intimate connection between the circadian cycle and the DNA damage checkpoints that is in part mediated by the Timeless protein. PMID- 15798198 TI - Stabilization and enhancement of the antiapoptotic activity of mcl-1 by TCTP. AB - Mcl-1 is one Bcl-2 family member that plays a pivotal role in animal development. The extremely labile nature of the Mcl-1 protein itself and the fact that the Mcl 1 level is a critical determinant in various cell survival pathways suggest that cellular processes that regulate Mcl-1 stability are as important as those that regulate Mcl-1 synthesis. Although transcriptional stimulation of Mcl-1 synthesis in response to various stimuli has been well documented, regulation of Mcl-1 stability has been hardly explored. In this study, we identified that the translationally controlled tumor protein (TCTP) was one cellular factor that interacted with Mcl-1 and modulated Mcl-1 stability. While overexpression of TCTP augmented the protein stability of Mcl-1, knockdown expression of TCTP by RNA interference destabilized Mcl-1. Furthermore, TCTP stabilized Mcl-1 through interfering with Mcl-1's degradation by the ubiquitin-dependent proteasome degradation pathway, and the TCTP binding-defective mutant of Mcl-1 (K257V) was much more susceptible to degradation and manifested a compromised antiapoptotic activity. Taken together, these results suggest that TCTP modulates Mcl-1's antiapoptotic activity by modulating its protein stability. The possible mechanism(s) involved in TCTP's modulation process is discussed. PMID- 15798200 TI - Wing-to-Leg homeosis by spineless causes apoptosis regulated by Fish-lips, a novel leucine-rich repeat transmembrane protein. AB - Growth, patterning, and apoptosis are mutually interactive during development. For example, cells that select an abnormal fate in a developing field are frequently removed by apoptosis. An important issue in this process that needs to be resolved is the mechanism used by cells to discern their correct fate from an abnormal fate. In order to examine this issue, we developed an animal model that expresses the dioxin receptor homolog Spineless (Ss) ectopically in the Drosophila wing. The presence of mosaic clones ectopically expressing ss results in a local transformation of organ identity, homeosis, from wing into a leg or antenna. The cells with misspecified fates subsequently activate c-Jun N-terminal kinase to undergo apoptosis in an autonomous or nonautonomous manner depending on their position within the wing, suggesting that a cell-cell interaction is, at least in some cases, involved in the detection of misspecified cells. Similar position dependence is commonly observed when various homeotic genes controlling the body segments are ectopically expressed. The autonomous and nonautonomous apoptosis caused by ss is regulated by a novel leucine-rich repeat family transmembrane protein, Fish-lips (Fili) that interacts with surrounding normal cells. These data support a mechanism in which the lack of some membrane proteins helps to recognize the presence of different cell types and direct these cells to an apoptotic fate in order to exclude them from the normal developing field. PMID- 15798199 TI - RAD-51-dependent and -independent roles of a Caenorhabditis elegans BRCA2-related protein during DNA double-strand break repair. AB - The BRCA2 tumor suppressor is implicated in DNA double-strand break (DSB) repair by homologous recombination (HR), where it regulates the RAD51 recombinase. We describe a BRCA2-related protein of Caenorhabditis elegans (CeBRC-2) that interacts directly with RAD-51 via a single BRC motif and that binds preferentially to single-stranded DNA through an oligonucleotide-oligosaccharide binding fold. Cebrc-2 mutants fail to repair meiotic or radiation-induced DSBs by HR due to inefficient RAD-51 nuclear localization and a failure to target RAD-51 to sites of DSBs. Genetic and cytological comparisons of Cebrc-2 and rad-51 mutants revealed fundamental phenotypic differences that suggest a role for Cebrc 2 in promoting the use of an alternative repair pathway in the absence of rad-51 and independent of nonhomologous end joining (NHEJ). Unlike rad-51 mutants, Cebrc 2 mutants also accumulate RPA-1 at DSBs, and abnormal chromosome aggregates that arise during the meiotic prophase can be rescued by blocking the NHEJ pathway. CeBRC-2 also forms foci in response to DNA damage and can do so independently of rad-51. Thus, CeBRC-2 not only regulates RAD-51 during HR but can also function independently of rad-51 in DSB repair processes. PMID- 15798201 TI - Akt-dependent cell size regulation by the adhesion molecule on glia occurs independently of phosphatidylinositol 3-kinase and Rheb signaling. AB - The role of cell adhesion molecules in mediating interactions with neighboring cells and the extracellular matrix has long been appreciated. More recently, these molecules have been shown to modulate intracellular signal transduction cascades critical for cell growth and proliferation. Expression of adhesion molecule on glia (AMOG) is downregulated in human and mouse gliomas, suggesting that AMOG may be important for growth regulation in the brain. In this report, we examined the role of AMOG expression on cell growth and intracellular signal transduction. We show that AMOG does not negatively regulate cell growth in vitro or in vivo. Instead, expression of AMOG in AMOG-deficient cells results in a dramatic increase in cell size associated with protein kinase B/Akt hyperactivation, which occurs independent of phosphatidylinositol 3-kinase activation. AMOG-mediated Akt phosphorylation specifically activates the mTOR/p70S6 kinase pathway previously implicated in cell size regulation, but it does not depend on tuberous sclerosis complex/Ras homolog enriched in brain (Rheb) signaling. These data support a novel role for a glial adhesion molecule in cell size regulation through selective activation of the Akt/mTOR/S6K signal transduction pathway. PMID- 15798202 TI - Inactivation of the arylhydrocarbon receptor nuclear translocator (Arnt) suppresses von Hippel-Lindau disease-associated vascular tumors in mice. AB - Patients with germ line mutations in the VHL tumor suppressor gene are predisposed to the development of highly vascularized tumors within multiple tissues. Loss of pVHL results in constitutive activation of the transcription factors HIF-1 and HIF-2, whose relative contributions to the pathogenesis of the VHL phenotype have yet to be defined. In order to examine the role of HIF in von Hippel-Lindau (VHL)-associated vascular tumorigenesis, we utilized Cre-loxP mediated recombination to inactivate hypoxia-inducible factor-1alpha (Hif-1alpha) and arylhydrocarbon receptor nuclear translocator (Arnt) genes in a VHL mouse model of cavernous liver hemangiomas and polycythemia. Deletion of Hif-1alpha did not affect the development of vascular tumors and polycythemia, nor did it suppress the increased expression of vascular endothelial growth factor (Vegf) and erythropoietin (Epo). In contrast, phosphoglycerokinase (Pgk) expression was substantially decreased, providing evidence for target gene-dependent functional redundancy between different Hif transcription factors. Inactivation of Arnt completely suppressed the development of hemangiomas, polycythemia, and Hif induced gene expression. Here, we demonstrate genetically that the development of VHL-associated vascular tumors in the liver depends on functional ARNT. Furthermore, we provide evidence that individual HIF transcription factors may play distinct roles in the development of specific VHL disease manifestations. PMID- 15798203 TI - Muscle-specific signaling mechanism that links actin dynamics to serum response factor. AB - Myocardin and the myocardin-related transcription factors (MRTFs) MRTF-A and MRTF B are coactivators for serum response factor (SRF), which regulates genes involved in cell proliferation, migration, cytoskeletal dynamics, and myogenesis. MRTF-A has been shown to translocate to the nucleus and activate SRF in response to Rho signaling and actin polymerization. Previously, we described a muscle specific actin-binding protein named striated muscle activator of Rho signaling (STARS) that also activates SRF through a Rho-dependent mechanism. Here we show that STARS activates SRF by inducing the nuclear translocation of MRTFs. The STARS-dependent nuclear import of MRTFs requires RhoA and actin polymerization, and the actin-binding domain of STARS is necessary and sufficient for this activity. A knockdown of endogenous STARS expression by using small interfering RNA significantly reduced SRF activity in differentiated C2C12 skeletal muscle cells and cardiac myocytes. The ability of STARS to promote the nuclear localization of MRTFs and SRF-mediated transcription provides a potential muscle specific mechanism for linking changes in actin dynamics and sarcomere structure with striated muscle gene expression. PMID- 15798204 TI - Oct-1 counteracts autoinhibition of Runx2 DNA binding to form a novel Runx2/Oct-1 complex on the promoter of the mammary gland-specific gene beta-casein. AB - The transcription factor Runx2 is essential for the expression of a number of bone-specific genes and is primarily considered a master regulator of bone development. Runx2 is also expressed in mammary epithelial cells, but its role in the mammary gland has not been established. Here we show that Runx2 forms a novel complex with the ubiquitous transcription factor Oct-1 to regulate the expression of the mammary gland-specific gene beta-casein. The Runx2/Oct-1 complex forms on a Runx/octamer element which is highly conserved in casein promoters. Chromatin immunoprecipitation, RNA interference, promoter mutagenesis, and transient expression analyses were used to demonstrate that the Runx2/Oct-1 complex contributes to the transcriptional regulation of the beta-casein gene. Analysis of the complex revealed autoinhibitory domains for DNA binding in both the N terminal and the C-terminal regions of Runx2. Oct-1 stimulates the recruitment of Runx2 to the beta-casein promoter by interacting with the C-terminal region of Runx2, suggesting that Oct-1 stimulates Runx2 recruitment by relieving the autoinhibition of Runx2 DNA binding. These findings demonstrate that Runx2 collaborates with Oct-1 and contributes to the expression of a mammary gland specific gene. PMID- 15798205 TI - Novel functions of the phospholipase D2-Phox homology domain in protein kinase Czeta activation. AB - It has been established that protein kinase Czeta (PKCzeta) participates in diverse signaling pathways and cellular functions in a wide variety of cells, exhibiting properties relevant to cellular survival and proliferation. Currently, however, the regulation mechanism of PKCzeta remains elusive. Here, for the first time, we determine that phospholipase D2 (PLD2) enhances PKCzeta activity through direct interaction in a lipase activity-independent manner. This interaction of the PLD2-Phox homology (PX) domain with the PKCzeta-kinase domain also induces the activation loop phosphorylation of PKCzeta and downstream signal stimulation, as measured by p70 S6 kinase phosphorylation. Furthermore, only the PLD2-PX domain directly stimulates PKCzeta activity in vitro, and it is necessary for the formation of the ternary complex with phosphoinositide-dependent kinase 1 and PKCzeta. The mutant that substitutes the triple lysine residues (Lys101, Lys102, and Lys103) within the PLD2-PX domain with alanine abolishes interaction with the PKCzeta-kinase domain and activation of PKCzeta. Moreover, breast cancer cell viability is significantly affected by PLD2 silencing. Taken together, these results suggest that the PLD2-mediated PKCzeta activation is induced by its PX domain performing both direct activation of PKCzeta and assistance of activation loop phosphorylation. Furthermore, we find it is an important factor in the survival of breast cancer cells. PMID- 15798206 TI - Histone dynamics on the interleukin-2 gene in response to T-cell activation. AB - Several models have been proposed for the mechanism of chromatin remodelling across the promoters of inducible genes in mammalian cells. The most commonly held model is one of cooccupation where histone proteins are modified by acetylation or phosphorylation and nucleosomes are remodelled, allowing the assembly of transcription factor complexes. Using chromatin immunoprecipitation, we observed an apparent decrease of histone acetylation and phosphorylation signals at the proximal promoter region of the inducible interleukin-2 and granulocyte-macrophage colony-stimulating factor genes in response to T-cell activation. We showed that this apparent decrease was due to a loss of histone H3 and H4 proteins corresponding to a decrease in nucleosome occupation of the promoter. This histone loss is reversible; it is dependent on the continual presence of appropriate activating signals and transcription factors and is not dependent on the acetylation status of the histone proteins. These data show for the first time that histone proteins are lost from a mammalian promoter upon activation of transcription and support a model of activation-dependent disassembly and reassembly of nucleosomes. PMID- 15798207 TI - Long-range interactions between three transcriptional enhancers, active Vkappa gene promoters, and a 3' boundary sequence spanning 46 kilobases. AB - The mouse immunoglobulin kappa (Igkappa) gene contains an intronic enhancer and two enhancers downstream of its transcription unit. Using chromosome conformation capture technology, we demonstrate that rearranged and actively transcribed Igkappa alleles in MPC-11 plasmacytoma cells exhibit mutual interactions over 22 kb between these three enhancers and Vkappa gene promoters. In addition, the 5' region of the active transcription unit exhibits a continuum of interactions with downstream chromatin segments. We also observe interactions between Ei and E3' with 3' boundary sequences 24 kb downstream of Ed, adjacent to a neighboring housekeeping gene. Very similar interactions between the enhancers are also exhibited by normal B cells isolated from mouse splenic tissue but not by germ line transcriptionally inactive alleles of T cells or P815 mastocytoma cells, which exhibit a seemingly linear chromatin organization. These results fit a looping mechanism for enhancer function like in the beta-globin locus and suggest a dynamic modulation of the spatial organization of the active Igkappa locus. Chromatin immunoprecipitation experiments reveal that the interacting Igkappa gene cis-acting sequences are associated with AP-4, E47, and p65NF-kappaB, potential protein candidates that may be responsible for initiating and/or maintaining the formation of these higher-order complexes. However, S107 plasmacytoma cells that lack NF-kappaB still exhibit mutual interactions between the Igkappa gene enhancers. PMID- 15798208 TI - Rhythmic serotonin N-acetyltransferase mRNA degradation is essential for the maintenance of its circadian oscillation. AB - Serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase [AANAT]) is the key enzyme in melatonin synthesis regulated by circadian rhythm. To date, our understanding of the oscillatory mechanism of melatonin has been limited to autoregulatory transcriptional and posttranslational regulations of AANAT mRNA. In this study, we identify three proteins from pineal glands that associate with cis-acting elements within species-specific AANAT 3' untranslated regions to mediate mRNA degradation. These proteins include heterogeneous nuclear ribonucleoprotein R (hnRNP R), hnRNP Q, and hnRNP L. Their RNA-destabilizing function was determined by RNA interference and overexpression approaches. Expression patterns of these factors in pineal glands display robust circadian rhythm. The enhanced levels detected after midnight correlate with an abrupt decline in AANAT mRNA level. A mathematical model for the AANAT mRNA profile and its experimental evidence with rat pinealocytes indicates that rhythmic AANAT mRNA degradation mediated by hnRNP R, hnRNP Q, and hnRNP L is a key process in the regulation of its circadian oscillation. PMID- 15798211 TI - Strong polyadenylation and weak pausing combine to cause efficient termination of transcription in the human Ggamma-globin gene. AB - The human gamma-globin genes form part of a 5-kb tandem duplication within the beta-globin gene cluster on chromosome 11. Despite a high degree of identity between the two genes, we show that while the upstream Ggamma-globin gene terminates transcription efficiently, termination in the Agamma gene is inefficient. This is primarily due to the different strengths of the polyA signals of the two genes; Ggamma-globin has a functionally stronger polyA signal than the Agamma gene. The probable cause of this difference in polyA efficiency characteristics lies with a number of base changes which reduce the G/U content of the GU/U-rich region of the Agamma polyA signal relative to that of Ggamma. The 3' flanking regions of the two gamma-globin genes have similar abilities to promote transcription termination. We found no evidence to suggest a cotranscriptional cleavage event, such as that seen in the human beta-globin gene, occurs in either gamma-globin 3' flank. Instead we find evidence that the 3' flank of the Ggamma-globin gene contains multiple weak pause elements which, combined with the strong polyA signal the gene possesses, are likely to cause gradual termination across the 3' flank. PMID- 15798209 TI - The p53 tumor suppressor protein represses human snRNA gene transcription by RNA polymerases II and III independently of sequence-specific DNA binding. AB - Human U1 and U6 snRNA genes are transcribed by RNA polymerases II and III, respectively. While the p53 tumor suppressor protein is a general repressor of RNA polymerase III transcription, whether p53 regulates snRNA gene transcription by RNA polymerase II is uncertain. The data presented herein indicate that p53 is an effective repressor of snRNA gene transcription by both polymerases. Both U1 and U6 transcription in vitro is repressed by recombinant p53, and endogenous p53 occupancy at these promoters is stimulated by UV light. In response to UV light, U1 and U6 transcription is strongly repressed. Human U1 genes, but not U6 genes, contain a high-affinity p53 response element located within the core promoter region. Nonetheless, this element is not required for p53 repression and mutant p53 molecules that do not bind DNA can maintain repression, suggesting a reliance on protein interactions for p53 promoter recruitment. Recruitment may be mediated by the general transcription factors TATA-box binding protein and snRNA activating protein complex, which interact well with p53 and function for both RNA polymerase II and III transcription. PMID- 15798210 TI - Myocyte enhancer factor 2 activates promoter sequences of the human AbetaH-J-J locus, encoding aspartyl-beta-hydroxylase, junctin, and junctate. AB - Alternative splicing of the locus AbetaH-J-J generates three functionally distinct proteins: an enzyme, AbetaH (aspartyl-beta-hydroxylase), a structural protein of the sarcoplasmic reticulum membrane (junctin), and an integral membrane calcium binding protein (junctate). Junctin and junctate are two important proteins involved in calcium regulation in eukaryotic cells. To understand the regulation of these two proteins, we identified and functionally characterized one of the two promoter sequences of the AbetaH-J-J locus. We demonstrate that the P2 promoter of the AbetaH-J-J locus contains (i) a minimal sequence localized within a region -159 bp from the transcription initiation site, which is sufficient to activate transcription of both mRNAs; (ii) sequences which bind known transcriptional factors such as those belonging to the myocyte enhancer factor 2 (MEF-2), MEF-3, and NF-kappaB protein families; and (iii) sequences bound by unknown proteins. The functional characterization of the minimal promoter in C2C12 cells and in the rat soleus muscle in vivo model indicates the existence of cis elements having positive and negative effects on transcription. In addition, our data demonstrate that in striated muscle cells the calcium-dependent transcription factor MEF-2 is crucial for the transcription activity directed by the P2 promoter. The transcription directed by the AbetaH-J J P2 promoter is induced by high expression of MEF-2, further stimulated by calcineurin and Ca2+/calmodulin-dependent protein kinase I, and inhibited by histone deacetylase 4. PMID- 15798212 TI - The SR protein SC35 is responsible for aberrant splicing of the E1alpha pyruvate dehydrogenase mRNA in a case of mental retardation with lactic acidosis. AB - Pyruvate dehydrogenase (PDH) complex deficiency is a major cause of lactic acidosis and Leigh's encephalomyelopathies in infancy and childhood, resulting in early death in the majority of patients. Most of the molecular defects have been localized in the coding regions of the E1alpha PDH gene. Recently, we identified a novel mutation of the E1alpha PDH gene in a patient with an encephalopathy and lactic acidosis. This mutation, located downstream of exon 7, activates a cryptic splice donor and leads to the retention of intronic sequences. Here, we demonstrate that the mutation results in an increased binding of the SR protein SC35. Consistently, ectopic overexpression of this splicing factor enhanced the use of the cryptic splice site, whereas small interfering RNA-mediated reduction of the SC35 protein levels in primary fibroblasts from the patient resulted in the almost complete disappearance of the aberrantly spliced E1alpha PDH mRNA. Our findings open the exciting prospect for a novel therapy of an inherited disease by altering the level of a specific splicing factor. PMID- 15798213 TI - Cotranscriptional recruitment of the pseudouridylsynthetase Cbf5p and of the RNA binding protein Naf1p during H/ACA snoRNP assembly. AB - H/ACA small nucleolar ribonucleoprotein particles (snoRNPs) are essential for the maturation and pseudouridylation of the precursor of rRNAs and other stable RNAs. Although the RNA and protein components of these RNPs have been identified, the mechanisms by which they are assembled in vivo are poorly understood. Here we show that the RNA binding protein Naf1p, which is required for H/ACA snoRNPs stability, associates with RNA polymerase II-associated proteins Spt16p, Tfg1p, and Sub1p and with H/ACA snoRNP proteins. Chromatin immunoprecipitation experiments show that Naf1p and the pseudouridylsynthetase Cbf5p cross-link specifically with the chromatin of H/ACA small nucleolar RNA (snoRNA) genes. Naf1p and Cbf5p cross-link predominantly with the 3' end of these genes, in a pattern similar to that observed for transcription elongation factor Spt16p. Cross-linking of Naf1p to H/ACA snoRNA genes requires active transcription and intact H/ACA snoRNA sequences but does not require the RNA polymerase II CTD kinase Ctk1p. These results suggest that Naf1p and Cbf5p are recruited in a cotranscriptional manner during H/ACA snoRNP assembly, possibly by binding to the nascent H/ACA snoRNA transcript during elongation or termination of transcription of H/ACA snoRNA genes. PMID- 15798214 TI - A novel domain in Set2 mediates RNA polymerase II interaction and couples histone H3 K36 methylation with transcript elongation. AB - Histone methylation and the enzymes that mediate it are important regulators of chromatin structure and gene transcription. In particular, the histone H3 lysine 36 (K36) methyltransferase Set2 has recently been shown to associate with the phosphorylated C-terminal domain (CTD) of RNA polymerase II (RNAPII), implying that this enzyme has an important role in the transcription elongation process. Here we show that a novel domain in the C terminus of Set2 is responsible for interaction between Set2 and RNAPII. This domain, termed the Set2 Rpb1 interacting (SRI) domain, is encompassed by amino acid residues 619 to 718 in Set2 and is found to occur in a number of putative Set2 homologs from Schizosaccharomyces pombe to humans. Unexpectedly, BIACORE analysis reveals that the SRI domain binds specifically, and with high affinity, to CTD repeats that are doubly modified (serine 2 and serine 5 phosphorylated), indicating that Set2 association across the body of genes requires a specific pattern of phosphorylated RNAPII. Deletion of the SRI domain not only abolishes Set2-RNAPII interaction but also abolishes K36 methylation in vivo, indicating that this interaction is required for establishing K36 methylation on chromatin. Using 6 azauracil (6AU) as an indicator of transcription elongation defects, we found that deletion of the SRI domain conferred a strong resistance to this compound, which was identical to that observed with set2 deletion mutants. Furthermore, yeast strains carrying set2 alleles that are catalytically inactive or yeast strains bearing point mutations at K36 were also found to be resistant to 6AU. These data suggest that it is the methylation by Set2 that affects transcription elongation. In agreement with this, we have determined that deletion of SET2, its SRI domain, or amino acid substitutions at K36 result in an alteration of RNAPII occupancy levels over transcribing genes. Taken together, these data indicate K36 methylation, established by the SRI domain-mediated association of Set2 with RNAPII, plays an important role in the transcription elongation process. PMID- 15798215 TI - YB-1 autoregulates translation of its own mRNA at or prior to the step of 40S ribosomal subunit joining. AB - YB-1 is a member of the numerous families of proteins with an evolutionary ancient cold-shock domain. It is involved in many DNA- and RNA-dependent events and regulates gene expression at different levels. Previously, we found a regulatory element within the 3' untranslated region (UTR) of YB-1 mRNA that specifically interacted with YB-1 and poly(A)-binding protein (PABP); we also showed that PABP positively affected YB-1 mRNA translation in a poly(A) tail independent manner (O. V. Skabkina, M. A. Skabkin, N. V. Popova, D. N. Lyabin, L. O. Penalva, and L. P. Ovchinnikov, J. Biol. Chem. 278:18191-18198, 2003). Here, YB-1 is shown to strongly and specifically inhibit its own synthesis at the stage of initiation, with accumulation of its mRNA in the form of free mRNPs. YB-1 and PABP binding sites have been mapped on the YB-1 mRNA regulatory element. These were UCCAG/ACAA for YB-1 and a approximately 50-nucleotide A-rich sequence for PABP that overlapped each other. PABP competes with YB-1 for binding to the YB-1 mRNA regulatory element and restores translational activity of YB-1 mRNA that has been inhibited by YB-1. Thus, YB-1 negatively regulates its own synthesis, presumably by specific interaction with the 3'UTR regulatory element, whereas PABP restores translational activity of YB-1 mRNA by displacing YB-1 from this element. PMID- 15798216 TI - p75-Ras-GRF1 is a c-Jun/AP-1 target protein: its up regulation results in increased Ras activity and is necessary for c-Jun-induced nonadherent growth of Rat1a cells. AB - The c-Jun/AP-1 transcription complex is associated with diverse cellular processes such as differentiation, proliferation, transformation, and apoptosis. These different biological endpoints are likely achieved by the regulation of specific target gene expression. We describe the identification of Ras guanine nucleotide exchange factor 1, Ras-GRF1, by microarray analysis as a c-Jun/AP-1 regulated gene essential for anchorage-independent growth of immortalized rat fibroblasts. Increased Ras-GRF1 expression, in response to inducible c-Jun expression in Rat1a fibroblasts, was confirmed by both real-time PCR and Northern blot analysis. We show that c-Jun/AP-1 can bind and activate the Ras-GRF1 promoter in vivo. A 75-kDa c-Jun/AP-1-inducible protein, p75-Ras-GRF1, was detected, and the inhibition of its expression with antisense oligomers significantly blocked c-Jun-regulated anchorage-independent cell growth. p75-Ras GRF1 expression occurred with a concomitant increase in activated Ras (GTP bound), and the activation of Ras was significantly inhibited by antisense Ras GRF1 oligomers. Moreover, p75-Ras-GRF1 could be coprecipitated with a Ras dominant-negative glutathione S-transferase (GST) construct, GST-Ras15A, demonstrating an interaction between p75-Ras-GRF1 and Ras. A downstream target of Ras activation, Elk-1, had increased transcriptional activity in c-Jun-expressing cells, and this activation was inhibited by dominant-negative Ras. In addition, c Jun overexpression resulted in an increase in phospho-AKT while phosphorylation of ERK1/2 remained largely unaffected. The inhibition of phosphatidylinositol 3 kinase (PI3K)-AKT signal transduction by Ly294002 and wortmannin significantly blocked c-Jun-regulated morphological transformation, while inhibition of basal MEK-ERK activity with PD98059 and U0126 had little effect. We conclude that c Jun/AP-1 regulates endogenous p75-Ras-GRF1 expression and that c-Jun/AP-1 regulated anchorage-independent cell growth requires activation of Ras-PI3K-AKT signal transduction. PMID- 15798217 TI - Transforming growth factor beta facilitates beta-TrCP-mediated degradation of Cdc25A in a Smad3-dependent manner. AB - Ubiquitin-dependent degradation of Cdc25A is a major mechanism for damage-induced S-phase checkpoint. Two ubiquitin ligases, the Skp1-cullin-beta-TrCP (SCFbeta TrCP) complex and the anaphase-promoting complex (APCCdh1), are involved in Cdc25A degradation. Here we demonstrate that the transforming growth factor beta (TGF-beta)-Smad3 pathway promotes SCF(beta-TrCP)-mediated Cdc25A ubiquitination. Cells treated with TGF-beta, as well as cells transfected with Smad3 or a constitutively active type I TGF-beta receptor, exhibit increased ubiquitination and markedly shortened half-lives of Cdc25A. Furthermore, Cdc25A is stabilized in cells transfected with Smad3 small interfering RNA (siRNA) and cells from Smad3 null mice. TGF-beta-induced ubiquitination is associated with Cdc25A phosphorylation at the beta-TrCP docking site (DS82G motif) and physical association of Cdc25A with Smad3 and beta-TrCP. Cdc25A mutant proteins deficient in DS82G phosphorylation are resistant to TGF-beta-Smad3-induced degradation, whereas a Cdc25A mutant protein defective in APCCdh1 recognition undergoes efficient degradation. Smad3 siRNA inhibits beta-TrCP-Cdc25A interaction and Cdc25A degradation in response to TGF-beta. beta-TrCP2 siRNA also inhibits Smad3 induced Cdc25A degradation. In contrast, Cdh1 siRNA had no effect on Cdc25A down regulation by Smad3. These data suggest that Smad3 plays a key role in the regulation of Cdc25A ubiquitination by SCFbeta-TrCP and that Cdc25A stabilization observed in various cancers could be associated with defects in the TGF-beta Smad3 pathway. PMID- 15798219 TI - Early embryonic lethality in mice with targeted deletion of the CTP:phosphocholine cytidylyltransferase alpha gene (Pcyt1a). AB - CTP:phosphocholine cytidylyltransferase (CCT) catalyzes a rate-controlling step in the biosynthesis of phosphatidylcholine (PtdCho). Multiple CCT isoforms, CCTalpha, CCTbeta2, and CCTbeta3, are encoded by two genes, Pcyt1a and Pcyt1b. The importance of CCTalpha in mice was investigated by deleting exons 5 and 6 in the Pcyt1a gene using the Cre-lox system. Pcyt1a-/- zygotes failed to form blastocysts, did not develop past embryonic day 3.5 (E3.5), and failed to implant. In situ hybridization in E11.5 embryos showed that Pcyt1a is expressed ubiquitously, with the highest level in fetal liver, and CCTalpha transcripts are significantly more abundant than transcripts encoding CCTbeta or phosphatidylethanolamine (PtdEtn) N-methyl transferase, two other enzymes capable of producing PtdCho. Reduction of the CCTalpha transcripts in heterozygous E11.5 embryos was accompanied by upregulation of CCTbeta and PtdEtn N-methyltransferase transcripts. In contrast, enzymatic and real-time PCR data revealed that CCTbeta (Pcyt1b) expression is not upregulated to compensate for the reduction in CCTalpha expression in adult liver and other tissues from Pcyt1a+/- heterozygous mice. PtdCho biosynthesis measured by choline incorporation into isolated hepatocytes was not compromised in the Pcyt1a+/- mice. Liver PtdCho mass was the same in Pcyt1a+/+ and Pcyt1a+/- adult animals, but lung PtdCho mass decreased in the heterozygous mice. These data show that CCTalpha expression is required for early embryonic development, but that a 50% reduction in enzyme activity has little detectable impact on the operation of the CDP-choline metabolic pathway in adult tissues. PMID- 15798218 TI - Posttranscriptional downregulation of c-IAP2 by the ubiquitin protein ligase c IAP1 in vivo. AB - Inhibitor of apoptosis proteins (IAPs) c-IAP1 and c-IAP2 were identified as part of the tumor necrosis factor receptor 2 (TNFR2) signaling complex and have been implicated as intermediaries in tumor necrosis factor alpha signaling. Like all RING domain-containing IAPs, c-IAP1 and c-IAP2 have ubiquitin protein ligase (E3) activity. To explore the function of c-IAP1 in a physiologic setting, c-IAP1 deficient mice were generated by homologous gene recombination. These animals are viable and have no obvious sensitization to proapoptotic stimuli. Cells from c IAP1(-/-) mice do, however, express markedly elevated levels of c-IAP2 protein in the absence of increased c-IAP2 mRNA. In contrast to reports implicating c-IAPs in the activation of NF-kappaB, resting and cytokine-induced NF-kappaB activation was not impaired in c-IAP1-deficient cells. Transient transfection studies with wild-type and E3-defective c-IAP1 revealed that c-IAP2 is a direct target for c IAP1-mediated ubiquitination and subsequent degradation, which are potentiated by the adaptor function of TRAF2. Thus, the c-IAPs represent a pair of TNFR associated ubiquitin protein ligases in which one regulates the expression of the other by a posttranscriptional and E3-dependent mechanism. PMID- 15798222 TI - Changes in the intracranial rheoencephalogram at lower limit of cerebral blood flow autoregulation. AB - Cerebral blood flow (CBF) reactivity monitoring is an appropriate primary parameter to evaluate cerebral resuscitation due to a systemic or regional cerebral injury leading to possible irreversible brain injury. Use of the electrical impedance method to estimate CBF is rare, as the method's anatomical background is not well understood. Use of intracranial rheoencephalography (iREG) during hemorrhage and comparison of iREG to other CBF measurements have not been previously reported. Our hypothesis was that iREG would reflect early cerebrovascular alteration (CBF autoregulation). Studies comparing iREG, laser Doppler flowmetry and ultrasound were undertaken on anesthetized rats to define CBF changes during hemorrhage. Blood was removed at a rate required to achieve a mean arterial blood pressure (MABP) of 40 mm Hg over 15 min. Estimation of CBF was taken with intracranial, bipolar REG (REG I; n=14), laser Doppler flowmetry (LDF; n=3) and carotid flow by ultrasound (n=11). Data were processed off-line. During the initial phase of hemorrhage, when MABP was close to 40 mm Hg, intracranial REG amplitude transiently increased (80.94%); LDF (77.92%) and carotid flow (52.04%) decreased and changed with systemic arterial pressure. Intracranial REG amplitude change suggests classical CBF autoregulation, demonstrating its close relationship to arteriolar changes. The studies indicate that iREG might reflect cerebrovascular responses more accurately than changes in local CBF measured by LDF and carotid flow. REG may indicate promise as a continuous, non-invasive life-sign monitoring tool with potential advantages over ultrasound, the CBF measurement technique normally applied in clinical practice. REG has particular advantages in non-hospital settings such as military and emergency medicine. PMID- 15798221 TI - Matrix metalloproteinases are not essential for aggrecan turnover during normal skeletal growth and development. AB - The growth plate is a transitional region of cartilage and highly diversified chondrocytes that controls long bone formation. The composition of growth plate cartilage changes markedly from the epiphysis to the metaphysis, notably with the loss of type II collagen, concomitant with an increase in MMP-13; type X collagen; and the C-propeptide of type II collagen. In contrast, the fate of aggrecan in the growth plate is not clear: there is biosynthesis and loss of aggrecan from hypertrophic cartilage, but the mechanism of loss is unknown. All matrix metalloproteinases (MMPs) cleave aggrecan between amino acids N341 and F342 in the proteinase-sensitive interglobular domain (IGD), and MMPs in the growth plate are thought to have a role in aggrecanolysis. We have generated mice with aggrecan resistant to proteolysis by MMPs in the IGD and found that the mice develop normally with no skeletal deformities. The mutant mice do not accumulate aggrecan, and there is no significant compensatory proteolysis occurring at alternate sites in the IGD. Our studies reveal that MMP cleavage in this key region is not a predominant mechanism for removing aggrecan from growth plate cartilage. PMID- 15798223 TI - Maxwell-Wagner relaxation in electrical imaging. AB - The electric field tomography (EFT) method exploits interaction of high-frequency electric field with an inhomogeneous conductive medium without contact with the electrodes. The interaction is accompanied by a high-frequency redistribution of free charges inside the medium and leads to small and regular phase shifts of the field in the area surrounding an object. Such a kind of phenomenon is referred to as the Maxwell-Wagner relaxation. Measuring the perturbations of the field using the set of electrodes placed around the object enables us to reconstruct the internal structure of the medium, generally the spatial distribution of a nonlinear combination of permittivity and resistivity. In the case of biomedical applications the result of measurements is determined mainly by the resistivity of the tissues. Three-dimensional simulation based on the finite element method has demonstrated the feasibility of the technique. PMID- 15798224 TI - Detection of emboli in vessels using electrical impedance measurements--phantom and electrodes. AB - A phantom was constructed to simulate the electrical properties of the neck. A range of possible electrode configurations was then examined in order to improve the sensitivity of the impedance measurement method for the in vivo detection of air emboli. The neck phantom consisted of simulated skin, fat and muscle layers made of agar and a conductive rubber tube mimicking the common carotid artery. The ring-shaped electrodes with a guard electrode showed the highest sensitivity to emboli at short distances. PMID- 15798225 TI - Prediction of mechanical properties of human trabecular bone by electrical measurements. AB - In trabecular bone, the interrelationships of electrical and dielectric properties with mechanical characteristics are poorly known. Information on these relations is crucial for evaluation of the diagnostic potential of impedance techniques. In this study, electrical and dielectric properties, i.e. permittivity, conductivity, phase angle, loss factor, specific impedance and dissipation factor of human trabecular bone samples (n=26, harvested from the distal femur and proximal tibia) were characterized in a wide frequency range (50 Hz-5 MHz). Mechanical properties, i.e. Young's modulus, ultimate strength, yield stress, yield strain and resilience of the samples (n=20) were determined by using destructive compressive testing. Subsequently, measurements of electrical and dielectric properties were repeated after mechanical testing. The measurements were also repeated for the control samples (n=6) that were not mechanically tested. Electrical, dielectric or mechanical properties showed no significant differences between the intact femoral and tibial samples. The electrical and dielectric parameters as well as the linear correlations between the dielectric and electrical parameters with mechanical parameters were strongly frequency dependent. At the frequency of 1.2 MHz, the relative permittivity showed the strongest linear correlations with the Young's modulus (r=0.71, p<0.01, n=20) and ultimate strength (r=0.73, p<0.01, n=20). Permittivity and dissipation factor showed statistically significant changes after mechanical testing. Our results suggest that the measurements of low frequency electrical and dielectric properties may provide information on the mechanical status of trabecular bone and, possibly, may even help to diagnose bone microdamage. In the future, these measurement techniques may be further developed for use during open surgery, such as bone grafting or total hip replacement surgery. PMID- 15798220 TI - Phosphorylation of p21 in G2/M promotes cyclin B-Cdc2 kinase activity. AB - Little is known about the posttranslational control of the cyclin-dependent protein kinase (CDK) inhibitor p21. We describe here a transient phosphorylation of p21 in the G2/M phase. G2/M-phosphorylated p21 is short-lived relative to hypophosphorylated p21. p21 becomes nuclear during S phase, prior to its phosphorylation by CDK2. S126-phosphorylated cyclin B1 binds to T57 phosphorylated p21. Cdc2 kinase activation is delayed in p21-deficient cells due to delayed association between Cdc2 and cyclin B1. Cyclin B1-Cdc2 kinase activity and G2/M progression in p21-/- cells are restored after reexpression of wild-type but not T57A mutant p21. The cyclin B1 S126A mutant exhibits reduced Cdc2 binding and has low kinase activity. Phosphorylated p21 binds to cyclin B1 when Cdc2 is phosphorylated on Y15 and associates poorly with the complex. Dephosphorylation on Y15 and phosphorylation on T161 promotes Cdc2 binding to the p21-cyclin B1 complex, which becomes activated as a kinase. Thus, hyperphosphorylated p21 activates the Cdc2 kinase in the G2/M transition. PMID- 15798227 TI - Phase angle is a predictor of basal metabolic rate in female patients with anorexia nervosa. AB - The aim of the study was to evaluate the relationship between basal metabolic rate (BMR) and bioelectrical impedance analysis (BIA) in undernourished female patients with anorexia nervosa. Participants were 86 female patients with anorexia nervosa (age 20.8+/-4.7 years; weight 39.3+/-5.2 kg; body mass index 15.4+/-1.6 kg m-2). BMR was measured by indirect calorimetry and single-frequency BIA was determined at 50 kHz on the whole body. The BIA variables considered were resistance, reactance, phase angle and the bioimpedance index (height2/resistance). Fat-free mass was calculated from subcutaneous skin fold thickness. In the study group BMR was 3782+/-661 kJ d-1 while bioimpedance index varied between 27.6 and 49.9 cm2 Omega-1 and phase angle between 2.54 degrees and 6.49 degrees. BMR was significantly correlated with weight, height, body mass index and fat-free mass, and, among BIA variables, with reactance and phase angle. Multiple regression analysis indicated that phase angle was a predictor of BMR not only when solely BIA variables were considered, but also in combination with either weight and age or fat-free mass. In conclusion, phase angle emerged as a strong predictor of BMR in female patients with anorexia nervosa. Nevertheless, further studies are necessary to confirm this finding in other forms of protein energy malnutrition and justify the inclusion of BIA variables in the equations used to predict BMR in the clinical setting. PMID- 15798226 TI - Body composition modeling in the calf using an equivalent circuit model of multi frequency bioimpedance analysis. AB - An equivalent electrical circuit model is used to describe the response of different tissue components in the calf to multi-frequency current. This model includes seven electrical components: skin resistance, contact capacitance, fat resistance, fat capacitance, extracellular resistance, intracellular resistance and cell membrane capacitance. Calf bioimpedance was measured on 30 pts using a multi-frequency bioimpedance device (Xitron 4200) with a range of frequency from 5 kHz to 1000 kHz. MRI was performed on each measured calf to provide body composition components: fat, muscle mass and bone. An equivalent circuit containing seven parameters (P1, P2, P3, P4, Q1, Q2, Q3) was constructed to represent the model. To identify the effect of different body compositions on their parameters, subjects were subgrouped according to (1) their range of fat mass: F1>0.4 kg, F2>0.4 & F2<0.25 kg and F3<0.25 kg; (2) their range of muscle mass: M1>1.2 kg, M2<1.2 & M2>1.0 kg and M3<0.25 kg. Curve fitting and simulation programs (Matlab Toolbox) were used to obtain the solution of the electrical equations. The results show a decrease in impedance with an increase in excitation frequency that differed among subjects with different fat contents. Simulation results show a high correlation (R2>0.98) between the bioimpedance measurements and the value calculated from the model. There are significant differences in parameters P1 (32.5+/-5.9 versus 26+/-4.4, p<0.05), P3 (-15,330+/ 3352 versus -10,973+/-3448, p<0.05) and P4 (42,640 versus 24,191, p<0.05) between groups F1 and F3. P2 is significantly different (1045+/-442 versus 1407+/-349, p<0.05) between groups M1 and M2. The parameters that characterize the bioimpedance data depend upon many more tissue characteristics of electrical properties than those incorporated in current models and they are affected by aspects of body composition that are not considered in the fitting of bioimpedance data. This study shows a new model and methodology to analyze bioimpedance data and further work is likely to lead to much better understanding of electrical properties of body tissue. PMID- 15798228 TI - Monitoring lung edema using the pacemaker pulse and skin electrodes. AB - Previous clinical studies have shown that impedance measurements using right ventricular (RV) leads can monitor congestion due to heart failure. We previously reported on a three-fold advantage of bipolar left ventricular (LV) leads, which are near the lung, over RV leads in detecting pulmonary edema with impedance. A combined system of internal and external electrodes is now investigated using computer models, for use with conventional cardiac resynchronization (CRT) systems with unipolar LV leads. The system uses the normal LV pacing pulse as current source, and the resultant voltage at two skin electrodes to obtain a lung edema impedance (Z) measurement. Using gated MRIs, thoracic computer models of 3.8 million control volumes were constructed. Changes of Z with edema were simulated with a conventional totally implanted system, as well as with combined implanted-external systems. Right atrial (RA), RV, RV defibrillator coil and LV leads were used. Per cent Z responses to edema were compared. The all implanted responses were RA: 11.8%, RV: 8.6%, RVcoil: 11.3%, LV: 23.8%. The combined system responses were LV-ext: 21.45%, RA-ext: 10.13%, LV-arm leg: 26.08%. The computer models suggest that combined internal-external systems can be as sensitive as the totally implanted ones. Lung edema may be monitored at follow up or home for LV paced patients with only two external electrodes. Using very low impedance configurations optimized by computer can greatly maximize the response, with a cost of poor stability. PMID- 15798229 TI - Bioimpedance dispersion width as a parameter to monitor living tissues. AB - In the case of living tissues, the spectral width of the electrical bioimpedance dispersions (closely related with the alpha parameter in the Cole equation) evolves during the ischemic periods. This parameter is often ignored in favor of other bioimpedance parameters such as the central frequency or the resistivity at low frequencies. The object of this paper is to analyze the significance of this parameter through computer simulations (in the alpha and beta dispersion regions) and to demonstrate its practical importance through experimental studies performed in rat kidneys during cold preservation. The simulations indicate that the dispersion width could be determined by the morphology of the extra-cellular spaces. The experimental studies show that it is a unique parameter able to detect certain conditions such as a warm ischemia period prior to cold preservation or the effect of a drug (Swinholide A) able to disrupt the cytoskeleton. The main conclusion is that, thanks to the alpha parameter in the Cole equation, the bioimpedance is not only useful to monitor the intra/extra cellular volume imbalances or the inter-cellular junctions resistance but also to detect tissue structural alterations. PMID- 15798230 TI - Automatic detection of detached and erroneous electrodes in electrical impedance tomography. AB - One unfortunate occurrence in experimental measurements with electrical impedance tomography is electrodes which become detached or poorly connected, such that the measured data cannot be used. This paper presents an automatic approach to detect such erroneous electrodes. It is based on the assumption that all valid measurements are related by the image reconstruction model, while the measurements from erroneous electrodes are unrelated. The method estimates the data at an electrode based on the measurements from all other electrodes, and compares it to the measurements. If these data match adequately, the set of electrodes does not contain an erroneous electrode. In order to detect an erroneous electrode amongst N electrodes, all sets of N-1 electrodes are tested, and the set with the best match between measurements and estimate is identified as the one which excludes the erroneous electrode. The method was tested on simulated and experimental data and showed consistent identification of erroneous electrodes with those made by experts. PMID- 15798232 TI - Design of electrode array for impedance measurement of lesions in arteries. AB - Use of impedance catheters can provide additional information about the composition and the morphology of early plaques in arteries. However, for a correct interpretation of the impedance data recorded inside a vessel, the extra vessel conditions should not influence the measurement results. In this paper, we estimate the influence of the extra-vessel conditions on the impedance measurement of a vessel wall by using FEM simulation and a two-layer model. Therefore sensitivity fields are simulated. The simulations are validated by experiments and compared to analytical solutions. Further, the influence of the inner radius of a vessel on the measurement result is determined by FEM simulations. From experiments based on the two-layer model, it is found that the apparent resistance depends on the thickness of the first layer and the separation distance of the electrode structure. The measured result corresponds to the results of the FEM simulations, whereas the analytical solution assuming point electrodes is different from the measurement and simulation results. Under the assumption of homogenous and linear volume conductors, the FEM simulated distributions of sensitivity fields are determined. The inner diameter of the artery has no influence on the measurement results. The FEM simulation can support the design of electrode configuration and geometries for impedance catheters. PMID- 15798231 TI - Excitation patterns in three-dimensional electrical impedance tomography. AB - Electrical impedance tomography (EIT) is a non-invasive technique that aims to reconstruct images of internal electrical properties of a domain, based on electrical measurements on the periphery. Improvements in instrumentation and numerical modeling have led to three-dimensional (3D) imaging. The availability of 3D modeling and imaging raises the question of identifying the best possible excitation patterns that will yield to data, which can be used to produce the best image reconstruction of internal properties. In this work, we describe our 3D finite element model of EIT. Through singular value decomposition as well as examples of reconstructed images, we show that for a homogenous female breast model with four layers of electrodes, a driving pattern where each excitation plane is a sinusoidal pattern out-of-phase with its neighboring plane produces better qualitative images. However, in terms of quantitative imaging an excitation pattern where all electrode layers are in phase produces better results. PMID- 15798233 TI - Spatial resolution improvement of 3D EIT images by the shrinking sLORETA-FOCUSS algorithm. AB - This paper describes the use of the shrinking sLORETA-FOCUSS algorithm to improve the spatial resolution of three-dimensional (3D) EIT images. Conventional EIT yields inaccurate, low spatial resolution images, due to noise, the low sensitivity of boundary voltages to inner conductivity perturbations and a limited number of boundary voltage measurements. The focal underdetermined system solver (FOCUSS) algorithm produces a localized energy solution based on the weighted minimum-norm least-squares (MNLS) solution. It was successfully applied for the spatial resolution improvement of EIT images of simulated and tank data for a 2D homogeneous circular disc. However, due to the fact that a 3D mesh system contains many more elements, much more memory is required to store the weighting matrix. In order to extend the work to 3D, the shrinking-FOCUSS method is utilized to shrink the solution space as well as the weighting matrix in each iteration step. The solution of the standardized low resolution electromagnetic tomography algorithm (sLORETA) is adopted as the initial estimate of the shrinking-FOCUSS. The effectiveness is verified by implementing the new algorithm on tank data for a three-dimensional homogeneous sphere. PMID- 15798234 TI - Using the GRID to improve the computation speed of electrical impedance tomography (EIT) reconstruction algorithms. AB - In our group at University College London, we have been developing electrical impedance tomography (EIT) of brain function. We have attempted to improve image quality by the use of realistic anatomical meshes and, more recently, non-linear reconstruction methods. Reconstruction with linear methods, with pre-processing, may take up to a few minutes per image for even detailed meshes. However, iterative non-linear reconstruction methods require much more computational resources, and reconstruction with detailed meshes was taking far too long for clinical use. We present a solution to this timing bottleneck, using the resources of the GRID, the development of coordinated computing resources over the internet that are not subject to centralized control using standard, open, general-purpose protocols and are transparent to the user. Optimization was performed by splitting reconstruction of image series into individual jobs of one image each; no parallelization was attempted. Using the GRID middleware 'Condor' and a cluster of 920 nodes, reconstruction of EIT images of the human head with a non-linear algorithm was speeded up by 25-40 times compared to serial processing of each image. This distributed method is of direct practical value in applications such as EIT of epileptic seizures where hundreds of images are collected over the few minutes of a seizure and will be of value to clinical data collection with similar requirements. In the future, the same resources could be employed for the more ambitious task of parallelized code. PMID- 15798236 TI - Cardiorespiratory monitoring equipment interferes with whole body impedance measurements. AB - Bioelectrical impedance measurements are widely used for the study of body composition. Commonly measurements are made at 50 kHz to estimate total body water or at low frequencies (<10 kHz) to estimate extracellular fluid volume. These measurements can be obtained as single measurements at discrete frequencies, or as fitted data interpolated from plots of measurements made at multiple frequencies. This study compared single frequency and multiple frequency (MF) measurements taken in the intensive care environment. MF bioimpedance (4 1000 kHz) was measured on an adult with and without cardiorespiratory monitoring, and on babies in the neonatal intensive care unit. Measurements obtained at individual frequencies were plotted against frequency and examined for the presence of outlying points. Fitted data for measurements obtained at 5 kHz and 50 kHz with and without cardiorespiratory monitoring were compared. Significant artefacts were detected in measurements at approximately 50 kHz and at integral divisions of this frequency as a result of interference from cardiorespiratory monitors. Single frequency measurements taken at these frequencies may be subject to errors that would be difficult to detect without the aid of information obtained from MF measurements. PMID- 15798235 TI - Dynamic electrical impedance imaging with the interacting multiple model scheme. AB - In this paper, an effective dynamical EIT imaging scheme is presented for on-line monitoring of the abruptly changing resistivity distribution inside the object, based on the interacting multiple model (IMM) algorithm. The inverse problem is treated as a stochastic nonlinear state estimation problem with the time-varying resistivity (state) being estimated on-line with the aid of the IMM algorithm. In the design of the IMM algorithm multiple models with different process noise covariance are incorporated to reduce the modeling uncertainty. Simulations and phantom experiments are provided to illustrate the proposed algorithm. PMID- 15798237 TI - Solution of the inverse problem of magnetic induction tomography (MIT). AB - Magnetic induction tomography (MIT) of biological tissue is used to reconstruct the changes in the complex conductivity distribution inside an object under investigation. The measurement principle is based on determining the perturbation DeltaB of a primary alternating magnetic field B0, which is coupled from an array of excitation coils to the object under investigation. The corresponding voltages DeltaV and V0 induced in a receiver coil carry the information about the passive electrical properties (i.e. conductivity, permittivity and permeability). The reconstruction of the conductivity distribution requires the solution of a 3D inverse eddy current problem. As in EIT the inverse problem is ill-posed and on this account some regularization scheme has to be applied. We developed an inverse solver based on the Gauss-Newton-one-step method for differential imaging, and we implemented and tested four different regularization schemes: the first and second approaches employ a classical smoothness criterion using the unit matrix and a differential matrix of first order as the regularization matrix. The third method is based on variance uniformization, and the fourth method is based on the truncated singular value decomposition. Reconstructions were carried out with synthetic measurement data generated with a spherical perturbation at different locations within a conducting cylinder. Data were generated on a different mesh and 1% random noise was added. The model contained 16 excitation coils and 32 receiver coils which could be combined pairwise to give 16 planar gradiometers. With 32 receiver coils all regularization methods yield fairly good 3D-images of the modelled changes of the conductivity distribution, and prove the feasibility of difference imaging with MIT. The reconstructed perturbations appear at the right location, and their size is in the expected range. With 16 planar gradiometers an additional spurious feature appears mirrored with respect to the median plane with negative sign. This demonstrates that a symmetrical arrangement with one ring of planar gradiometers cannot distinguish between a positive conductivity change at the true location and a negative conductivity change at the mirrored location. PMID- 15798239 TI - Design and performance of a planar-array MIT system with normal sensor alignment. AB - In this study the performance of a planar array for magnetic induction tomography (MIT) was investigated and the results of measurements to determine the precision and sensitivity of the sensor were undertaken. A planar-array MIT system utilizing flux-linkage minimization for the primary field has been constructed and evaluated. The system comprises 4 printed excitation coils of 4 turns which were shielded, 8 surface-mount inductors of inductance 10 microH as sensor, mounted such that in principle no primary-field flux threads them, and a calibration coil to produce a strong primary field. The excitation current was multiplexed via relays to drive the excitation and reference coils. The noise values were similar in real and imaginary components in the lower frequencies and the factor to which the primary field could be reduced was greatest in the nearest coil. Methods for determining the true real and imaginary components and for flux-linkage minimization for the primary field for variations in channel sensitivities are described and the results of measurements of the system's noise and drift are given. A SNR of 47 dB was observed at 4 MHz when a 0.3 Sm-1 saline filled tank of dimensions 20 cmx20 cmx10 cm was placed centrally over the array. Finally, images were reconstructed from measurements of saline samples in a free space background, with the samples moved past the array in 21 1 cm steps to emulate mechanical scanning of the array. The image reconstruction characteristics of the planar array in conjunction with the reconstruction technique employed are discussed. PMID- 15798238 TI - Generating accurate finite element meshes for the forward model of the human head in EIT. AB - The use of realistic anatomy in the model used for image reconstruction in EIT of brain function appears to confer significant improvements compared to geometric shapes such as a sphere. Accurate model geometry may be achieved by numerical models based on magnetic resonance images (MRIs) of the head, and this group has elected to use finite element meshing (FEM) as it enables detailed internal anatomy to be modelled and has the capability to incorporate information about tissue anisotropy. In this paper a method for generating accurate FEMs of the human head is presented where MRI images are manually segmented using custom adaptation of industry standard commercial design software packages. This is illustrated with example surface models and meshes from adult epilepsy patients, a neonatal baby and a phantom latex tank incorporating a real skull. Mesh quality is assessed in terms of element stretch and hence distortion. PMID- 15798240 TI - Effect of lower body negative pressure and gravity on regional lung ventilation determined by EIT. AB - The aim of our study was to check the effect of varying blood volume in the chest and gravity on the distribution of ventilation and aeration in the lungs. The change in intrathoracic blood volume was elicited by application of lower body negative pressure (LBNP) of -50 cmH2O. The variation of gravity in terms of hypogravity (approximately 0g) and hypergravity (approximately 2g) was induced by changes in vertical acceleration achieved during parabolic flights. Local ventilation magnitude and end-expiratory lung volume were determined in eight human subjects in the ventral and dorsal lung regions within a transverse cross section of the lower chest by electrical impedance tomography. The subjects were studied in a 20 degrees head-down tilted supine body position during tidal breathing and full forced expirations. During tidal breathing, a significant effect of gravity on local magnitude of ventilation and end-expiratory lung volume was detected in the dorsal lung regions both with and without LBNP. In the ventral regions, this gravity dependency was only observed during LBNP. During forced expiration, LBNP had almost no effect on local ventilation and end expiratory lung volume in either lung region. Gravity significantly influenced the end-expiratory lung volumes in dorsal lung regions. The results indicate that exposure to LBNP exerts a less appreciable effect on regional lung ventilation than the acute changes in gravity. PMID- 15798241 TI - Electrical conductivity images of biological tissue phantoms in MREIT. AB - We present cross-sectional conductivity images of two biological tissue phantoms. Each of the cylindrical phantoms with both diameter and height of 140 mm contained chunks of biological tissues such as bovine tongue and liver, porcine muscle and chicken breast within a conductive agar gelatin as the background medium. We attached four recessed electrodes on the sides of the phantom with equal spacing among them. Injecting current pulses of 480 or 120 mA ms into the phantom along two different directions, we measured the z-component Bz of the induced magnetic flux density B=(Bx, By, Bz) with a magnetic resonance electrical impedance tomography (MREIT) system based on a 3.0 T MRI scanner. Using the harmonic Bz algorithm, we reconstructed cross-sectional conductivity images from the measured Bz data. Reconstructed images clearly distinguish different tissues in terms of both their shapes and conductivity values. In this paper, we experimentally demonstrate the feasibility of the MREIT technique in producing conductivity images of different biological soft tissues with a high spatial resolution and accuracy when we use a sufficient amount of the injection current. PMID- 15798242 TI - Induced current magnetic resonance-electrical impedance tomography. AB - Magnetic resonance-electrical impedance tomography (MR-EIT) is a conductivity imaging method based on injecting currents into the object. In this study, a new MR-EIT method, whereby currents are induced inside the object by using external coils, is proposed. This new method is called induced current magnetic resonance electrical impedance tomography. In induced current MR-EIT surface electrodes are not used and thereby artifacts due to electrodes are eliminated. The reconstruction algorithm is based on the measurement of only one component of the secondary magnetic flux density. The algorithm is an iterative one, is 3D and is based on the solution of a linear matrix equation at each iteration. For the measurement of secondary magnetic flux density, a pulse sequence to be used in the MRI system is proposed. Numerical simulations are performed to test the algorithm for both noise-free and noisy cases. The singular value behavior of the matrix is monitored and it is observed that at least two current induction profiles improve the images significantly. It is shown that induced current MR EIT can be used to reconstruct absolute conductivity images without the need for any additional peripheral voltage measurement. PMID- 15798243 TI - A new type of gradiometer for the receiving circuit of magnetic induction tomography (MIT). AB - Magnetic induction tomography (MIT) is a low-resolution imaging modality which aims at the three-dimensional (3D) reconstruction of the electrical conductivity in objects from alternating magnetic fields. In MIT systems the magnetic field perturbations to be detected are very small when compared to the excitation field (ppm range). The voltage which is induced by the excitation field in the receiver coils must be suppressed for providing sufficient dynamic range. In the past, two very efficient strategies were proposed: adjusted planar gradiometers (PGRAD) and the orientation of a receiver coil with respect to the excitation coil such that the net magnetic flow is zero (zero flow coil, ZFC). In contrast to the PGRAD no voltage is induced in the ZFC by the main field. This is advantageous because two comparatively high voltages in the two gradiometer coils can never be subtracted perfectly, thus leaving a residual voltage which is prone to drift. However, a disadvantage of the ZFC is the higher susceptibility to interferences from far RF sources. In contrast, in the gradiometer such interferences are cancelled to a high degree. We developed a new type of gradiometer (zero flow gradiometer, ZFGRAD) which combines the advantages of ZFC and PGRAD. All three systems were compared with respect to sensitivity and perturbation to signal ratio (PSR) defined as the ratio of the signal change due to a magnetic perturbation field at the carrier frequency and the signal change due to shifting a metallic sphere between two test points. The spatial sensitivity of the three systems was found to be very similar. The PSR of the ZFGRAD was more than 12 times lower than that of the ZFC. Finally, the feasibility of image reconstruction with two arrays of eight excitation coils and eight ZFGRAD, respectively, was shown with a single step Gauss-Newton reconstructor and simulated measurement data generated for a cylindrical tank with a spherical perturbation. The resulting images show a clear, bright feature at the correct position of the sphere and are comparable to those with PGRAD arrays. PMID- 15798245 TI - Electrical bioimpedance readings increase with higher pressure applied to the measuring probe. AB - Electrical bioimpedance spectroscopy (EBIS) is a technique that uses a probe to calculate the transfer impedance from tissues. This transfer impedance can give information about the normal or pathological condition of the tissue. To take readings, pressure has to be applied to the probe in order to get a good contact between the electrodes and the tissue. We have been using EBIS to investigate the early diagnosis of dysplasia and cancer in the human cervix, oesophagus and bladder. We have found that, with increasing pressure (range used here was approximately 1 kPa to approximately 50 kPa), the resistivity readings increase in a consistent way up to 80%. In this paper, we show how this is a case in three different tissue types (oesophageal, gastric and vesical samples). These increases can be higher than those associated with the pathological changes that we are investigating (non-inflamed columnar tissue, for instance, shows values 50% higher than dysplastic columnar tissue). Finite-element modelling was also used to investigate the effect of volume reduction in the connective tissue or stroma. This simulation suggests no strong correlation between reduction of this structure and increase in resistivity. We hypothesize therefore that these changes may be mainly associated with the squeezing of water from the extracellular space. Finally, as pressure is difficult to control by hand, we raise the issue of the necessity of considering this variable when making EIS measurements. PMID- 15798244 TI - A comparison of sensors for minimizing the primary signal in planar-array magnetic induction tomography. AB - In magnetic induction tomography reducing the influence of the primary excitation field on the sensors can provide a significant improvement in SNR and/or allow the operating frequency to be reduced. For the purposes of imaging, it would be valuable if all, or a useful subset, of the detection coils could be rendered insensitive to the primary field for any excitation coil activated. Suitable schemes which have been previously suggested include the use of axial gradiometers and coil-orientation methods (Bx sensors). This paper examines the relative performance of each method through computer simulation of the sensitivity profiles produced by a single sensor, and comparison of reconstructed images produced by sensor arrays. A finite-difference model was used to determine the sensitivity profiles obtained with each type of sensor arrangement. The modelled volume was a cuboid of dimensions 50 cmx50 cmx12 cm with a uniform conductivity of 1 S m-1. The excitation coils were of 5 cm diameter and the detection coils of 5 mm diameter. The Bx sensors provided greater sensitivity than the axial gradiometers at all depths, other than on the surface layer of the volume. Images produced using a single-planar array were found to contain distortion which was reduced by the addition of a second array. PMID- 15798246 TI - Impedance changes in liver tissue exposed in vitro to high-energy ultrasound. AB - The present study reports the impedance changes observed in bovine liver samples exposed in vitro to high-intensity ultrasound. The measurement frequency ranged from 80 kHz to 2 MHz. The treatment resulted in the average increase of 20% in impedance magnitude at low frequency and the average decrease of 30% at high frequency. The phase angle increased significantly by more than 15 degrees at all measurement frequencies. The slope of the log-modulus of impedance against log frequency increased in treated tissue at frequencies above 500 kHz. This change was attributed to the alteration of the capacitive response of the tissue. The experimental observations are consistent with the known changes induced by high energy ultrasound in liver tissue. This study confirmed that ultrasound energy produces measurable changes in a tissue's impedance and that indices can be derived to distinguish between original and treated tissues. The results obtained in liver tissue need confirmation in organs treatable with therapeutic ultrasound, such as breast and prostate. PMID- 15798247 TI - Optimum electrode configuration for detection of leg movement using bio impedance. AB - Impedances and joint angles were simultaneously measured during ankle and knee movements. The correlation coefficients of the joint angle and the impedance change from human leg movement were obtained using an electro-goniometer and a four-channel impedance measurement system. Because the impedance changes resulting from ankle and knee movements depended heavily on the electrode placement, we determined the optimum electrode configurations for those movements by searching for high correlation coefficients, large impedance changes and minimum interferences in ten subjects (age: 20+/-4). Our optimum electrode configurations showed strong relationships between the ankle joint angle and lower leg impedance (correlation coefficient=-0.91+/-0.06) and between the knee joint angle and knee impedance (correlation coefficient=0.94+/-0.04). The reproducibilities of the impedance changes of five subjects due to the ankle and knee were 6.3+/-1.9% and 5.1+/-1.7% for the optimum electrode pairs, respectively. We propose that this optimum electrode configuration would be useful for future studies involving the convenient measurement of leg movements by the impedance method. PMID- 15798249 TI - EIT images of ventilation: what contributes to the resistivity changes? AB - One promising application of electrical impedance tomography (EIT) is the monitoring of pulmonary ventilation and edema. Using three-dimensional (3D) finite difference human models as virtual phantoms, the factors that contribute to the observed lung resistivity changes in the EIT images were investigated. The results showed that the factors included not only tissue resistivity or vessel volume changes, but also chest expansion and tissue/organ movement. The chest expansion introduced artifacts in the center of the EIT images, ranging from -2% to 31% of the image magnitude. With the increase of simulated chest expansion, the percentage contribution of chest expansion relative to lung resistivity change in the EIT image remained relatively constant. The averaged resistivity changes in the lung regions caused by chest expansion ranged from 0.65% to 18.31%. Tissue/organ movement resulted in an increased resistivity in the lung region and in the center anterior region of EIT images. The increased resistivity with inspiration observed in the heart region was caused mainly by a drop in the heart position, which reduced the heart area at the electrode level and was replaced by the lung tissue with higher resistivity. This study indicates that for the analysis of EIT, data errors caused by chest expansion and tissue/organ movement need to be considered. PMID- 15798248 TI - A six-ring probe for monitoring conductivity changes. AB - This paper presents the construction of a six-ring probe for monitoring immittance changes. The spatial sensitivity of the probe is defined. This is used to examine the uniqueness of the probe in terms of its application to monitoring conductivity changes. A spatial distribution of the sensitivity is presented for isotropic and anisotropic cases. The latter case is restricted only to anisotropy met when measuring muscles, i.e. diagonal anisotropy. Theoretical calculations performed using the finite element method were verified experimentally using a specially developed measuring system. An example of in vivo measurements is included. PMID- 15798250 TI - Effect of body mass index (BMI) on estimation of extracellular volume (ECV) in hemodialysis (HD) patients using segmental and whole body bioimpedance analysis. AB - The aim of the study was to investigate whether body mass index (BMI) influences the estimation of extracellular volume (ECV) in hemodialysis (HD) patients when using segmental bioimpedance analysis (SBIA) compared to wrist-to-ankle bioimpedance analysis (WBIA) during HD with ultrafiltration (UF). Twenty five HD patients (M:F 19:6,) were studied, and further subdivided into two groups of patients, one group with a high BMI (25 kg m-2) and the other with a low BMI (<25 kg m-2). Segmental (arm, trunk, leg) and wrist-to-ankle bioimpedance measurements on each patient were performed using a modified Xitron 4000B system (Xitron Technologies, San Diego, CA). No differences in extracellular resistance (R(E), ohms) between wrist-to-ankle (R(W)) and sum of segments (R(S)) were noted for either the high BMI (489.2+/-82 ohm versus 491.6+/-82 ohm, p=ns) or low BMI groups (560.8+/-77 ohm versus 557.5+/-75 ohm, p=ns). UF volume (UFV, liters) did not differ significantly between the groups (4.0+/-0.9 L versus 3.3+/-1.0 L, p=ns), but change in ECV (DeltaECV) differed not only between methods: WBIA versus SBIA in the high BMI group (2.74+/-1.1 L versus 3.64+/-1.4 L, p<0.001) and in the low BMI group (1.86+/-0.9 L versus 2.91+/-1.0 L, p<0.05) but also between the high and lower BMI groups with WBIA (2.74+/-1.1 L versus 1.86+/-0.9 L, p<0.01). However, there was no significant difference in SBIA between BMI groups. This study suggests that the segmental bioimpedance approach may more accurately reflect changes in ECV during HD with UF than whole body impedance measurements. PMID- 15798251 TI - Monte Carlo simulation of large electron fields. AB - Accurate simulation of large electron fields may lead to improved accuracy in Monte Carlo treatment planning while simplifying the commissioning procedure. We have used measurements made with wide-open jaws and no electron applicator to adjust simulation parameters. Central axis depth dose curves and profiles of 6-21 MeV electron beams measured in this geometry were used to estimate source and geometry parameters, including those that affect beam symmetry: incident beam direction and offset of the secondary scattering foil and monitor chamber from the beam axis. Parameter estimation relied on a comprehensive analysis of the sensitivity of the measured quantities, in the large field, to source and geometry parameters. Results demonstrate that the EGS4 Monte Carlo system is capable of matching dose distributions in the largest electron field to the least restrictive of 1 cGy or 1 mm, with D(max) of 100 cGy, over the full energy range. This match results in an underestimation of the bremsstrahlung dose of 10-20% at 15-21 MeV, exceeding the combined experimental and calculational uncertainty in this quantity of 3%. The simulation of electron scattering at energies of 15-21 MeV in EGS4 may be in error. The recently released EGSnrc/BEAMnrc system may provide a better match to measurement. PMID- 15798252 TI - Dual scattering foil design for poly-energetic electron beams. AB - The laser wakefield acceleration (LWFA) mechanism can accelerate electrons to energies within the 6-20 MeV range desired for therapy application. However, the energy spectrum of LWFA-generated electrons is broad, on the order of tens of MeV. Using existing laser technology, the therapeutic beam might require a significant energy spread to achieve clinically acceptable dose rates. The purpose of this work was to test the assumption that a scattering foil system designed for a mono-energetic beam would be suitable for a poly-energetic beam with a significant energy spread. Dual scattering foil systems were designed for mono-energetic beams using an existing analytical formalism based on Gaussian multiple-Coulomb scattering theory. The design criterion was to create a flat beam that would be suitable for fields up to 25 x 25 cm2 at 100 cm from the primary scattering foil. Radial planar fluence profiles for poly-energetic beams with energy spreads ranging from 0.5 MeV to 6.5 MeV were calculated using two methods: (a) analytically by summing beam profiles for a range of mono-energetic beams through the scattering foil system, and (b) by Monte Carlo using the EGS/BEAM code. The analytic calculations facilitated fine adjustments to the foil design, and the Monte Carlo calculations enabled us to verify the results of the analytic calculation and to determine the phase-space characteristics of the broadened beam. Results showed that the flatness of the scattered beam is fairly insensitive to the width of the input energy spectrum. Also, results showed that dose calculated by the analytical and Monte Carlo methods agreed very well in the central portion of the beam. Outside the useable field area, the differences between the analytical and Monte Carlo results were small but significant, possibly due to the small angle approximation. However, these did not affect the conclusion that a scattering foil system designed for a mono-energetic beam will be suitable for a poly-energetic beam with the same central energy. Further studies of the dosimetric properties of LWFA-generated electron beams will be done using Monte Carlo methods. PMID- 15798253 TI - Sensitivity of large-field electron beams to variations in a Monte Carlo accelerator model. AB - Adjustments made to Monte Carlo models during the commissioning of the simulation should be physically realistic and correspond to actual machine characteristics. Large electron fields, with the jaws fully open and the applicator removed, are sensitive to important source and geometry parameters and may provide the most accurate beam models, including those collimated by an applicator. We report on the results of a comprehensive Monte Carlo sensitivity study documenting the response of these large fields to changes in the configuration of a Siemens Primus linear accelerator. The study was performed for 6, 9 12, 15, 18 and 21 MeV configurations, and included variations of thickness, position and lateral alignment of all treatment head components. Variations of electron beam characteristics were also included in the study. Results were classified by their impact on central-axis depth dose distributions, including the bremsstrahlung tail, and on beam profiles near D(max) and in the bremsstrahlung region. Low energy results show an increased sensitivity to electron beam properties. High energy bremsstrahlung profiles are shown to be useful in determining misalignments between the beam axis and mechanical isocentre. For all energies, the alignment of the secondary scattering foil and monitor chamber are shown to be critical for correctly modelling beam asymmetries. The results suggest a methodology for commissioning of electron beams using Monte Carlo treatment head simulation. PMID- 15798254 TI - Experimental determination of electron source parameters for accurate Monte Carlo calculation of large field electron therapy. AB - Extensive work has been performed to validate Monte Carlo models for both photon and electron beams under standard conditions. However, for large field electron beam therapy, Monte Carlo simulations have not been able to provide good agreement when compared to the measured dose distributions. Since the accuracy of the calculation relies heavily on the geometry parameters of the linear accelerator and the characteristics of the incident electron beam, it is crucial to have a complete comprehension of these independent factors. In this work, the electron focal spot size for a CL21EX linac with various energies (6, 9, 12 and 16 MeV) was measured with a slit camera composed of alternating lead and paper sheets. For all the energies investigated, the electron focal spot is found to be elliptical and has a full width at half maximum (FWHM) ranging from 1.69 mm to 2.24 mm. A shift with respect to the crosshair was associated with each measured focal spot. In addition, we present an improved result for the large field in-air profile by utilizing a proposed divergent beam model in conjunction with the experimental focal spot dimension. This model can potentially provide a solution to the Monte Carlo validation of large field electron beam therapy. PMID- 15798255 TI - Development of a Monte Carlo model for the Brainlab microMLC. AB - Stereotactic radiosurgery with several static conformal beams shaped by a micro multileaf collimator (microMLC) is used to treat small irregularly shaped brain lesions. Our goal is to perform Monte Carlo calculations of dose distributions for certain treatment plans as a verification tool. A dedicated microMLC component module for the BEAMnrc code was developed as part of this project and was incorporated in a model of the Varian CL2300 linear accelerator 6 MV photon beam. As an initial validation of the code, the leaf geometry was visualized by tracing particles through the component module and recording their position each time a leaf boundary was crossed. The leaf dimensions were measured and the leaf material density and interleaf air gap were chosen to match the simulated leaf leakage profiles with film measurements in a solid water phantom. A comparison between Monte Carlo calculations and measurements (diode, radiographic film) was performed for square and irregularly shaped fields incident on flat and homogeneous water phantoms. Results show that Monte Carlo calculations agree with measured dose distributions to within 2% and/or 1 mm except for field size smaller than 1.2 cm diameter where agreement is within 5% due to uncertainties in measured output factors. PMID- 15798256 TI - The influence of beam model differences in the comparison of dose calculation algorithms for lung cancer treatment planning. AB - In this study, we show that beam model differences play an important role in the comparison of does calculated with various algorithms for lung cancer treatment planning. These differences may impact the accurate correlation of dose with clinical outcome. To accomplish this, we modified the beam model penumbral parameters in an equivalent path length (EPL) algorithm and subsequently compared the EPL doses with those generated with Monte Carlo (MC). A single AP beam was used for beam fitting. Two different beam models were generated for EPL calculations: (1) initial beam model (init_fit) and (2) optimized beam model (best_fit) , with parameters optimized to produce the best agreement with MC calculated profiles at several depths in a water phantom. For the 6 MV, AP beam, EPL(init_fit) calculations were on average within 2%/2 mm (1.4 mm max.) agreement with MC; the agreement for EPL(best_fit) was 2%/1.0 mm (1.3 mm max.) for EPL(best_fit). Treatment planning was performed using a realistic lung phantom using 6 and 15 MV photons. In all homogeneous phantom plans, EPL(best_fit) calculations were in better agreement with MC. In the heterogeneous 6 MV plan, differences between EPL(best_fit and init_fit) and MC were significant for the tumour. The EPL(init_fit), unlike the EPL(best_fit) calculation, showed large differences in the lung relative to MC. For the 15 MV heterogeneous plan, clinically important differences were found between EPL(best_fit or init_fit) and MC for tumour and lung, suggesting that the algorithmic difference in inhomogeneous cases, differences between EPL(best_fit) and MC for lung tissues were smaller compared to those between EPL(init_fit) and MC. Although the extent to which beam model differences impact the dose comparisons will be dependent upon beam parameters (orientation, field size and energy), and the size and location of the tumour, this study shows that failing to correctly account for beam model differences will lead to biased comparisons between dose algorithms. This may ultimately hinder our ability to accurately correlate dose with clinical outcome. PMID- 15798257 TI - Head-and-neck IMRT treatments assessed with a Monte Carlo dose calculation engine. AB - IMRT is frequently used in the head-and-neck region, which contains materials of widely differing densities (soft tissue, bone, air-cavities). Conventional methods of dose computation for these complex, inhomogeneous IMRT cases involve significant approximations. In the present work, a methodology for the development, commissioning and implementation of a Monte Carlo (MC) dose calculation engine for intensity modulated radiotherapy (MC-IMRT) is proposed which can be used by radiotherapy centres interested in developing MC-IMRT capabilities for research or clinical evaluations. The method proposes three levels for developing, commissioning and maintaining a MC-IMRT dose calculation engine: (a) development of a MC model of the linear accelerator, (b) validation of MC model for IMRT and (c) periodic quality assurance (QA) of the MC-IMRT system. The first step, level (a), in developing an MC-IMRT system is to build a model of the linac that correctly predicts standard open field measurements for percentage depth-dose and off-axis ratios. Validation of MC-IMRT, level (b), can be performed in a rando phantom and in a homogeneous water equivalent phantom. Ultimately, periodic quality assurance of the MC-IMRT system is needed to verify the MC-IMRT dose calculation system, level (c). Once the MC-IMRT dose calculation system is commissioned it can be applied to more complex clinical IMRT treatments. The MC-IMRT system implemented at the Royal Marsden Hospital was used for IMRT calculations for a patient undergoing treatment for primary disease with nodal involvement in the head-and-neck region (primary treated to 65 Gy and nodes to 54 Gy), while sparing the spinal cord, brain stem and parotid glands. Preliminary MC results predict a decrease of approximately 1-2 Gy in the median dose of both the primary tumour and nodal volumes (compared with both pencil beam and collapsed cone). This is possibly due to the large air-cavity (the larynx of the patient) situated in the centre of the primary PTV and the approximations present in the dose calculation. PMID- 15798258 TI - The importance of accurate linear accelerator head modelling for IMRT Monte Carlo calculations. AB - Two Monte Carlo dose engines for radiotherapy treatment planning, namely a beta release of Peregrine and MCDE (Monte Carlo dose engine), were compared with Helax TMS (collapsed cone superposition convolution) for a head and neck patient for the Elekta SLi plus linear accelerator. Deviations between the beta release of Peregrine and MCDE up to 10% were obtained in the dose volume histogram of the optical chiasm. It was illustrated that the differences are not caused by the particle transport in the patient, but by the modelling of the Elekta SLi plus accelerator head and more specifically the multileaf collimator (MLC). In MCDE two MLC modules (MLCQ and MLCE) were introduced to study the influence of the tongue-and-groove geometry, leaf bank tilt and leakage on the actual dose volume histograms. Differences in integral dose in the optical chiasm up to 3% between the two modules have been obtained. For single small offset beams though the FWHM of lateral profiles obtained with MLCE can differ by more than 1.5 mm from profiles obtained with MLCQ. Therefore, and because the recent version of MLCE is as fast as MLCQ, we advise to use MLCE for modelling the Elekta MLC. Nevertheless there still remains a large difference (up to 10%) between Peregrine and MCDE. By studying small offset beams we have shown that the profiles obtained with Peregrine are shifted, too wide and too flat compared with MCDE and phantom measurements. The overestimated integral doses for small beam segments explain the deviations observed in the dose volume histograms. The Helax-TMS results are in better agreement with MCDE, although deviations exceeding 5% have been observed in the optical chiasm. Monte Carlo dose deviations of more than 10% as found with Peregrine are unacceptable as an influence on the clinical outcome is possible and as the purpose of Monte Carlo treatment planning is to obtain an accuracy of 2%. We would like to emphasize that only the Elekta MLC has been tested in this work, so it is certainly possible that alpha releases of Peregrine provide more accurate results for other accelerators. PMID- 15798259 TI - Monte Carlo based modulated electron beam treatment planning using a few-leaf electron collimator--feasibility study. AB - Energy modulated electron beam therapy with conventional clinical accelerators has lagged behind photon IMRT despite its potential to achieve highly conformal dose distributions in superficial targets. One of the reasons for this is the absence of an automated collimating device that allows for the flexible delivery of a series of variable field openings. Electron-specific multileaf collimators attached to the bottom of the applicator require the use of a large number of motors and suffer from being relatively bulky and impractical for head and neck sites. In this work, we investigate the treatment planning aspects of a proposed 'few-leaf' electron collimator (FLEC) that consists of four motor-driven trimmer bars at the end of the applicator. The device is designed to serve as an accessory to standard equipment and allows for the shaping of any irregular field by combination of rectangular fieldlets. Using a Monte Carlo model of the FLEC, dose distributions are optimized using a simulated annealing (SA) inverse planning algorithm based on a limited number of Monte Carlo pre-generated, realistic phantom-specific dose kernels and user-specified dose-volume constraints. Using a phantom setup with an artificial target enclosed by organs at risk (OAR) as well as using a realistic patient case, we demonstrate that highly conformal distributions can be generated. Estimates of delivery times are made and show that a full treatment fraction can be kept to 15 min or less. PMID- 15798260 TI - Monte Carlo- versus pencil-beam-/collapsed-cone-dose calculation in a heterogeneous multi-layer phantom. AB - The aim of this work was to evaluate the accuracy of dose predicted in heterogeneous media by a pencil beam (PB), a collapsed cone (CC) and a Monte Carlo (MC) algorithm. For this purpose, a simple multi-layer phantom composed of Styrofoam and white polystyrene was irradiated with 10 x 10 cm2 as well as 20 x 20 cm2 open 6 MV photon fields. The beam axis was aligned parallel to the layers and various field offsets were applied. Thereby, the amount of lateral scatter was controlled. Dose measurements were performed with an ionization chamber positioned both in the central layer of white polystyrene and the adjacent layers of Styrofoam. It was found that, in white polystyrene, both MC and CC calculations agreed satisfactorily with the measurements whereas the PB algorithm calculated 12% higher doses on average. By studying off-axis dose profiles the observed differences in the calculation results increased dramatically for the three algorithms. In the regions of low density CC calculated 10% (8%) lower doses for the 10 x 10 cm2 (20 x 20 cm2) fields than MC. The MC data on the other hand agreed well with the measurements, presuming that proper replacement correction for the ionization chamber embedded in Styrofoam was performed. PB results evidently did not account for the scattering geometry and were therefore not really comparable. Our investigations showed that the PB algorithm generates very large errors for the dose in the vicinity of interfaces and within low density regions. We also found that for the used CC algorithm large deviations for the absolute dose (dose/monitor unit) occur in regions of electronic disequilibrium. The performance might be improved by better adapted parameters. Therefore, we recommend a careful investigation of the accuracy for dose calculations in heterogeneous media for each beam data set and algorithm. PMID- 15798261 TI - Dosimetric verification of IMRT treatment planning using Monte Carlo simulations for prostate cancer. AB - The purpose of this work is to investigate the accuracy of dose calculation of a commercial treatment planning system (Corvus, Normos Corp., Sewickley, PA). In this study, 30 prostate intensity-modulated radiotherapy (IMRT) treatment plans from the commercial treatment planning system were recalculated using the Monte Carlo method. Dose-volume histograms and isodose distributions were compared. Other quantities such as minimum dose to the target (D(min)), the dose received by 98% of the target volume (D98), dose at the isocentre (D(iso)), mean target dose (D(mean)) and the maximum critical structure dose (D(max)) were also evaluated based on our clinical criteria. For coplanar plans, the dose differences between Monte Carlo and the commercial treatment planning system with and without heterogeneity correction were not significant. The differences in the isocentre dose between the commercial treatment planning system and Monte Carlo simulations were less than 3% for all coplanar cases. The differences on D98 were less than 2% on average. The differences in the mean dose to the target between the commercial system and Monte Carlo results were within 3%. The differences in the maximum bladder dose were within 3% for most cases. The maximum dose differences for the rectum were less than 4% for all the cases. For non-coplanar plans, the difference in the minimum target dose between the treatment planning system and Monte Carlo calculations was up to 9% if the heterogeneity correction was not applied in Corvus. This was caused by the excessive attenuation of the non-coplanar beams by the femurs. When the heterogeneity correction was applied in Corvus, the differences were reduced significantly. These results suggest that heterogeneity correction should be used in dose calculation for prostate cancer with non-coplanar beam arrangements. PMID- 15798262 TI - IMRT head and neck treatment planning with a commercially available Monte Carlo based planning system. AB - The PEREGRINE Monte Carlo dose-calculation system (North American Scientific, Cranberry Township, PA) is the first commercially available Monte Carlo dose calculation code intended specifically for intensity modulated radiotherapy (IMRT) treatment planning and quality assurance. In order to assess the impact of Monte Carlo based dose calculations for IMRT clinical cases, dose distributions for 11 head and neck patients were evaluated using both PEREGRINE and the CORVUS (North American Scientific, Cranberry Township, PA) finite size pencil beam (FSPB) algorithm with equivalent path-length (EPL) inhomogeneity correction. For the target volumes, PEREGRINE calculations predict, on average, a less than 2% difference in the calculated mean and maximum doses to the gross tumour volume (GTV) and clinical target volume (CTV). An average 16% +/- 4% and 12% +/- 2% reduction in the volume covered by the prescription isodose line was observed for the GTV and CTV, respectively. Overall, no significant differences were noted in the doses to the mandible and spinal cord. For the parotid glands, PEREGRINE predicted a 6% +/- 1% increase in the volume of tissue receiving a dose greater than 25 Gy and an increase of 4% +/- 1% in the mean dose. Similar results were noted for the brainstem where PEREGRINE predicted a 6% +/- 2% increase in the mean dose. The observed differences between the PEREGRINE and CORVUS calculated dose distributions are attributed to secondary electron fluence perturbations, which are not modelled by the EPL correction, issues of organ outlining, particularly in the vicinity of air cavities, and differences in dose reporting (dose to water versus dose to tissue type). PMID- 15798263 TI - Effect of statistical uncertainties on Monte Carlo treatment planning. AB - This paper reviews the effect of statistical uncertainties on radiotherapy treatment planning using Monte Carlo simulations. We discuss issues related to the statistical analysis of Monte Carlo dose calculations for realistic clinical beams using various variance reduction or time saving techniques. We discuss the effect of statistical uncertainties on dose prescription and monitor unit calculation for conventional treatment and intensity-modulated radiotherapy (IMRT) based on Monte Carlo simulations. We show the effect of statistical uncertainties on beamlet dose calculation and plan optimization for IMRT and other advanced treatment techniques such as modulated electron radiotherapy (MERT). We provide practical guidelines for the clinical implementation of Monte Carlo treatment planning and show realistic examples of Monte Carlo based IMRT and MERT plans. PMID- 15798265 TI - Adjoint Monte Carlo method for prostate external photon beam treatment planning: an application to 3D patient anatomy. AB - Recently, the theoretical framework of the adjoint Monte Carlo (AMC) method has been developed using a simplified patient geometry. In this study, we extended our previous work by applying the AMC framework to a 3D anatomical model called VIP-Man constructed from the Visible Human images. First, the adjoint fluxes for the prostate (PTV) and rectum and bladder (organs at risk (OARs)) were calculated on a spherical surface of 1 m radius, centred at the centre of gravity of PTV. An importance ratio, defined as the PTV dose divided by the weighted OAR doses, was calculated for each of the available beamlets to select the beam angles. Finally, the detailed doses in PTV and OAR were calculated using a forward Monte Carlo simulation to include the electron transport. The dose information was then used to generate dose volume histograms (DVHs). The Pinnacle treatment planning system was also used to generate DVHs for the 3D plans with beam angles obtained from the AMC (3D-AMC) and a standard six-field conformal radiation therapy plan (3D CRT). Results show that the DVHs for prostate from 3D-AMC and the standard 3D-CRT are very similar, showing that both methods can deliver prescribed dose to the PTV. A substantial improvement in the DVHs for bladder and rectum was found for the 3D-AMC method in comparison to those obtained from 3D-CRT. However, the 3D AMC plan is less conformal than the 3D-CRT plan because only bladder, rectum and PTV are considered for calculating the importance ratios. Nevertheless, this study clearly demonstrated the feasibility of the AMC in selecting the beam directions as a part of a treatment planning based on the anatomical information in a 3D and realistic patient anatomy. PMID- 15798264 TI - A comparison of Monte Carlo dose calculation denoising techniques. AB - Recent studies have demonstrated that Monte Carlo (MC) denoising techniques can reduce MC radiotherapy dose computation time significantly by preferentially eliminating statistical fluctuations ('noise') through smoothing. In this study, we compare new and previously published approaches to MC denoising, including 3D wavelet threshold denoising with sub-band adaptive thresholding, content adaptive mean-median-hybrid (CAMH) filtering, locally adaptive Savitzky-Golay curve fitting (LASG), anisotropic diffusion (AD) and an iterative reduction of noise (IRON) method formulated as an optimization problem. Several challenging phantom and computed-tomography-based MC dose distributions with varying levels of noise formed the test set. Denoising effectiveness was measured in three ways: by improvements in the mean-square-error (MSE) with respect to a reference (low noise) dose distribution; by the maximum difference from the reference distribution and by the 'Van Dyk' pass/fail criteria of either adequate agreement with the reference image in low-gradient regions (within 2% in our case) or, in high-gradient regions, a distance-to-agreement-within-2% of less than 2 mm. Results varied significantly based on the dose test case: greater reductions in MSE were observed for the relatively smoother phantom-based dose distribution (up to a factor of 16 for the LASG algorithm); smaller reductions were seen for an intensity modulated radiation therapy (IMRT) head and neck case (typically, factors of 2-4). Although several algorithms reduced statistical noise for all test geometries, the LASG method had the best MSE reduction for three of the four test geometries, and performed the best for the Van Dyk criteria. However, the wavelet thresholding method performed better for the head and neck IMRT geometry and also decreased the maximum error more effectively than LASG. In almost all cases, the evaluated methods provided acceleration of MC results towards statistically more accurate results. PMID- 15798266 TI - Sensitivity study for CT image use in Monte Carlo treatment planning. AB - An important step in Monte Carlo treatment planning (MCTP), which is commonly performed uncritically, is segmentation of the patient CT data into a voxel phantom for dose calculation. In addition to assigning mass densities to voxels, as is done in conventional TP, this entails assigning media. Mis-assignment of media can potentially lead to significant dose errors in MCTP. In this work, a test phantom with exact-known composition was used to study CT segmentation errors and to quantify subsequent MCTP inaccuracies. For our test cases, we observed dose errors in some regions of up to 10% for 6 and 15 MV photons, more than 30% for an 18 MeV electron beam and more than 40% for 250 kVp photons. It is concluded that a careful CT calibration with a suitable phantom is essential. Generic calibrations and the use of commercial CT phantoms have to be critically assessed. PMID- 15798267 TI - Monte Carlo treatment planning for molecular targeted radiotherapy within the MINERVA system. AB - The aim of this project is to extend accurate and patient-specific treatment planning to new treatment modalities, such as molecular targeted radiation therapy, incorporating previously crafted and proven Monte Carlo and deterministic computation methods. A flexible software environment is being created that allows planning radiation treatment for these new modalities and combining different forms of radiation treatment with consideration of biological effects. The system uses common input interfaces, medical image sets for definition of patient geometry and dose reporting protocols. Previously, the Idaho National Engineering and Environmental Laboratory (INEEL), Montana State University (MSU) and Lawrence Livermore National Laboratory (LLNL) had accrued experience in the development and application of Monte Carlo based, three dimensional, computational dosimetry and treatment planning tools for radiotherapy in several specialized areas. In particular, INEEL and MSU have developed computational dosimetry systems for neutron radiotherapy and neutron capture therapy, while LLNL has developed the PEREGRINE computational system for external beam photon-electron therapy. Building on that experience, the INEEL and MSU are developing the MINERVA (modality inclusive environment for radiotherapeutic variable analysis) software system as a general framework for computational dosimetry and treatment planning for a variety of emerging forms of radiotherapy. In collaboration with this development, LLNL has extended its PEREGRINE code to accommodate internal sources for molecular targeted radiotherapy (MTR), and has interfaced it with the plugin architecture of MINERVA. Results from the extended PEREGRINE code have been compared to published data from other codes, and found to be in general agreement (EGS4-2%, MCNP-10%) (Descalle et al 2003 Cancer Biother. Radiopharm. 18 71-9). The code is currently being benchmarked against experimental data. The interpatient variability of the drug pharmacokinetics in MTR can only be properly accounted for by image-based, patient-specific treatment planning, as has been common in external beam radiation therapy for many years. MINERVA offers 3D Monte Carlo-based MTR treatment planning as its first integrated operational capability. The new MINERVA system will ultimately incorporate capabilities for a comprehensive list of radiation therapies. In progress are modules for external beam photon-electron therapy and boron neutron capture therapy (BNCT). Brachytherapy and proton therapy are planned. Through the open application programming interface (API), other groups can add their own modules and share them with the community. PMID- 15798269 TI - Monte Carlo dose calculations for spot scanned proton therapy. AB - Density heterogeneities can have a profound effect on dose distributions for proton therapy. Although analytical calculations in homogeneous media are relatively straightforward, the modelling of the propagation of the beam through density heterogeneities can be more problematical. At the Paul Scherrer Institute, an in-house dedicated Monte Carlo (MC) code has been used for over a decade to assess the possible deficiencies of the analytical calculations in patient geometries. The MC code has been optimized for speed, and as such traces primary protons only through the treatment nozzle and patient's CT. Contributions from nuclear interactions are modelled analytically with no tracing of secondary particles. The MC code has been verified against measured data in water and experimental proton radiographs through a heterogeneous anthropomorphic phantom. In comparison to the analytical calculation, the MC code has been applied to both spot scanned and intensity modulated proton therapy plans, and to a number of cases containing titanium metal implants. In summary, MC-based dose calculations could provide an invaluable tool for independently verifying the calculated dose distribution within a patient geometry as part of a comprehensive quality assurance protocol for proton treatment plans. PMID- 15798268 TI - Microionization chamber for reference dosimetry in IMRT verification: clinical implications on OAR dosimetric errors. AB - Intensity modulated radiotherapy (IMRT) has become a treatment of choice in many oncological institutions. Small fields or beamlets with sizes of 1 to 5 cm2 are now routinely used in IMRT delivery. Therefore small ionization chambers (IC) with sensitive volumes 0.1 cm3 are generally used for dose verification of an IMRT treatment. The measurement conditions during verification may be quite different from reference conditions normally encountered in clinical beam calibration, so dosimetry of these narrow photon beams pertains to the so-called non-reference conditions for beam calibration. This work aims at estimating the error made when measuring the organ at risk's (OAR) absolute dose by a micro ion chamber (microIC) in a typical IMRT treatment. The dose error comes from the assumption that the dosimetric parameters determining the absolute dose are the same as for the reference conditions. We have selected two clinical cases, treated by IMRT, for our dose error evaluations. Detailed geometrical simulation of the microIC and the dose verification set-up was performed. The Monte Carlo (MC) simulation allows us to calculate the dose measured by the chamber as a dose averaged over the air cavity within the ion-chamber active volume (D(air)). The absorbed dose to water (D(water)) is derived as the dose deposited inside the same volume, in the same geometrical position, filled and surrounded by water in the absence of the ion chamber. Therefore, the D(water)/D(air) dose ratio is the MC estimator of the total correction factor needed to convert the absorbed dose in air into the absorbed dose in water. The dose ratio was calculated for the microIC located at the isocentre within the OARs for both clinical cases. The clinical impact of the calculated dose error was found to be negligible for the studied IMRT treatments. PMID- 15798270 TI - 4D Monte Carlo simulation of proton beam scanning: modelling of variations in time and space to study the interplay between scanning pattern and time-dependent patient geometry. AB - When dosimetric effects in time-dependent geometries are studied, usually either the results of individual three-dimensional (3D) calculations are combined or probability-based approaches are applied. These methods may become cumbersome and time-consuming if high time resolution is required or if the geometry is complex. Furthermore, it is difficult to study double-dynamic systems, e.g., to investigate the influence of time-dependent beam delivery (i.e., magnetically moving beam spots in proton beam scanning) on the dose deposition in a moving target. We recently introduced the technique of 4D Monte Carlo dose calculation to model continuously changing geometries. In intensity modulated proton therapy, dose is delivered by individual pristine Bragg curves. Dose spots are positioned in the patient by varying magnetic field and beam energy. If the movement of these dose spots occurs during significant respiratory motion, interplay effects can take place. Because of the inhomogeneity of individual subfields, the consequences of motion can be more severe than in conventional proton therapy. We demonstrate how the technique of 4D Monte Carlo can be used to study interplay effects in proton beam scanning. Time-dependent beam delivery to a changing patient geometry is simulated in a single 4D dose calculation. Interplay effects between respiratory motion and beam scanning speed are demonstrated. PMID- 15798271 TI - Assigning nonelastic nuclear interaction cross sections to Hounsfield units for Monte Carlo treatment planning of proton beams. AB - In high energy clinical proton beams nonelastic nuclear interactions contribute substantially to the total dose. It is therefore of importance to know these contributions quantitatively and to be able to scale them correctly as a function of Hounsfield units obtained from CT data. In this work, the second of these issues has been addressed. The importance of taking material-dependent nonelastic nuclear interactions into account has been investigated for Monte Carlo calculations. A scaling curve for nonelastic nuclear interactions as a function of Hounsfield unit has been established and compared with similar data for the stopping powers. Monte Carlo simulations using McPTRAN.MEDIA and MCNPX have been performed in homogeneous media and in inhomogeneous slab geometries. The results show that for skeletal tissues and for adipose tissue, the tissue to water nonelastic cross section ratios differ up to 10% compared to the tissue to water stopping power ratios. This results in errors of the order of 2-3% when both contributions to the total dose are scaled in the same way (with stopping power ratios). Monte Carlo simulations in slab geometries with tissue materials for 200 MeV protons show similar effects, but when both contributions are scaled correctly the errors are not larger than 0.5% in the situations investigated here. PMID- 15798272 TI - A particle track-repeating algorithm for proton beam dose calculation. AB - A particle track-repeating algorithm has been developed for proton beam dose calculation for radiotherapy. Monoenergetic protons with 250 MeV kinetic energy were simulated in an infinite water phantom using the GEANT3 Monte Carlo code. The changes in location, angle and energy for every transport step and the energy deposition along the track were recorded for the primary protons and all secondary particles. When calculating dose for a patient with a realistic proton beam, the pre-generated particle tracks were repeated in the patient geometry consisting of air, soft tissue and bone. The medium and density for each dose scoring voxel in the patient geometry were derived from patient CT data. The starting point, at which a proton track was repeated, was determined according to the incident proton energy. Thus, any protons with kinetic energy less than 250 MeV can be simulated. Based on the direction of the incident proton, the tracks were first rotated and for the subsequent steps, the scattering angles were simply repeated for air and soft tissue but adjusted properly based on the scattering power for bone. The particle step lengths were adjusted based on the density for air and soft tissue and also on the stopping powers for bone while keeping the energy deposition unchanged in each step. The difference in nuclear interactions and secondary particle generation between water and these materials was ignored. The algorithm has been validated by comparing the dose distributions in uniform water and layered heterogeneous phantoms with those calculated using the GEANT3 code for 120, 150, 180 and 250 MeV proton beams. The differences between them were within 2%. The new algorithm was about 13 times faster than the GEANT3 Monte Carlo code for a uniform phantom geometry and over 700 times faster for a heterogeneous phantom geometry. PMID- 15798273 TI - The Monte Carlo SRNA-VOX code for 3D proton dose distribution in voxelized geometry using CT data. AB - This paper describes the application of the SRNA Monte Carlo package for proton transport simulations in complex geometry and different material compositions. The SRNA package was developed for 3D dose distribution calculation in proton therapy and dosimetry and it was based on the theory of multiple scattering. The decay of proton induced compound nuclei was simulated by the Russian MSDM model and our own using ICRU 63 data. The developed package consists of two codes: the SRNA-2KG, which simulates proton transport in combinatorial geometry and the SRNA VOX, which uses the voxelized geometry using the CT data and conversion of the Hounsfield's data to tissue elemental composition. Transition probabilities for both codes are prepared by the SRNADAT code. The simulation of the proton beam characterization by multi-layer Faraday cup, spatial distribution of positron emitters obtained by the SRNA-2KG code and intercomparison of computational codes in radiation dosimetry, indicate immediate application of the Monte Carlo techniques in clinical practice. In this paper, we briefly present the physical model implemented in the SRNA package, the ISTAR proton dose planning software, as well as the results of the numerical experiments with proton beams to obtain 3D dose distribution in the eye and breast tumour. PMID- 15798274 TI - Calculations of dose distributions using a neural network model. AB - The main goal of external beam radiotherapy is the treatment of tumours, while sparing, as much as possible, surrounding healthy tissues. In order to master and optimize the dose distribution within the patient, dosimetric planning has to be carried out. Thus, for determining the most accurate dose distribution during treatment planning, a compromise must be found between the precision and the speed of calculation. Current techniques, using analytic methods, models and databases, are rapid but lack precision. Enhanced precision can be achieved by using calculation codes based, for example, on Monte Carlo methods. However, in spite of all efforts to optimize speed (methods and computer improvements), Monte Carlo based methods remain painfully slow. A newer way to handle all of these problems is to use a new approach in dosimetric calculation by employing neural networks. Neural networks (Wu and Zhu 2000 Phys. Med. Biol. 45 913-22) provide the advantages of those various approaches while avoiding their main inconveniences, i.e., time-consumption calculations. This permits us to obtain quick and accurate results during clinical treatment planning. Currently, results obtained for a single depth-dose calculation using a Monte Carlo based code (such as BEAM (Rogers et al 2003 NRCC Report PIRS-0509(A) rev G)) require hours of computing. By contrast, the practical use of neural networks (Mathieu et al 2003 Proceedings Journees Scientifiques Francophones, SFRP) provides almost instant results and quite low errors (less than 2%) for a two-dimensional dosimetric map. PMID- 15798275 TI - Clinical use of a commercial Monte Carlo treatment planning system for electron beams. AB - In 2002 we fully implemented clinically a commercial Monte Carlo based treatment planning system for electron beams. The software, developed by MDS Nordion (presently Nucletron), is based on Kawrakow's VMC++ algorithm. The Monte Carlo module is integrated with our Theraplan Plustrade mark treatment planning system. An extensive commissioning process preceded clinical implementation of this software. Using a single virtual 'machine' for each electron beam energy, we can now calculate very accurately the dose distributions and the number of MU for any arbitrary field shape and SSD. This new treatment planning capability has significantly impacted our clinical practice. Since we are more confident of the actual dose delivered to a patient, we now calculate accurate three-dimensional (3D) dose distributions for a greater variety of techniques and anatomical sites than we have in the past. We use the Monte Carlo module to calculate dose for head and neck, breast, chest wall and abdominal treatments with electron beams applied either solo or in conjunction with photons. In some cases patient treatment decisions have been changed, as compared to how such patients would have been treated in the past. In this paper, we present the planning procedure and some clinical examples. PMID- 15798276 TI - Personal photon dosemeter trial--Devonport Royal Dockyard. AB - To establish an understanding of the operational responses of various personal dosemeters employed at Devonport and to assess new types of dosemeters, a photon dosemeter trial was conducted. Most day-to-day exposure is to relatively low dose rates. Therefore the suitability of each dosemeter for use within the relatively low Devonport dose rate environment has been assessed. The Panasonic TLD demonstrated a good representation of the dose within the medium to higher gamma energy ranges with an unexpected under-response at lower energies. The optically stimulated luminescent dosemeter showed a varied response within a degraded (60)Co environment. With consistent under-response, the NRPB TLD and film badge were found to generally be unsuitable for sites such as Devonport. The Harshaw TLD demonstrated a good representation of the dose. The RADOS RAD80 and QFD fail to meet current best industry standards. However, the QFD is the only direct reading dosemeter suitable for use where intrinsically safe equipment is required. The RADOS RAD52 and SAIC PD 2i show a reasonable representation of the dose received but should be adjusted to read within an operational (60)Co environment. Direct ion storage and Thermo [corrected] electronic personal dosemeters showed good representations of the dose. Inherent characteristics combined with the associated systems led to the conclusion that these dosemeters should be employed for preference. PMID- 15798277 TI - Reference biospheres for post-closure performance assessment: inter-comparison of SHETRAN simulations and BIOMASS results. AB - Example Reference Biosphere 2B (ERB2B) is a hypothetical river catchment, described in the IAEA-sponsored BIOMASS study on biosphere aspects of post closure radiological safety assessments for repositories for solid radioactive wastes. In ERB2B, a radioactively contaminated aquifer interacts with the soils and sediments of the river catchment. A 'semi-distributed', lumped-parameter model (SDLP) was set up for the site as part of the BIOMASS study. In the model, empirically derived transfer functions are used to reduce the complexity of real hydrological transport systems to readily calculable mass-balance accounting routines. In this work, a physically based, spatially distributed modelling system SHETRAN was set up for the site and comparison made with the existing SDLP model. The work has shown that, using standard soil properties in SHETRAN, the soil rapidly saturates and much of the hydrologically effective rainfall (precipitation less evapotranspiration) is lost as saturation-excess surface runoff. This is contrary to the assumptions in the SDLP model. The difficulty arose from the original formulation of catchment characteristics in BIOMASS. Specifically, there was a large water volume entering the soils from precipitation together with an upward flux of groundwater across the lower boundary of a substantial part of the catchment. This water had to be lost from the catchment in some way and the thinness of the soil zone precluded dominance of subsurface, lateral flow over surface runoff. Increasing the saturated conductivity from 1 to 20 m d(-1) reduced the surface flows to similar values to those assumed in the SDLP model (this could also have been achieved by increasing the soil depth). Even with the high saturated conductivity there were still major differences between the two representations. In the woodland on the upper slopes of the valley, the SHETRAN simulation was slightly wetter than the SDLP model, whereas in the shrubland and marshland near the river it was drier than the SDLP model. In the SDLP model, subsurface lateral flows are ignored if there is surface flow, and deep subsurface flows are ignored if there are shallow subsurface flows. In the SDLP model, there is a major assumed change in flow regime between summer and winter. This is not the case in the SHETRAN simulation. Overall, this work illustrates the problems of using 'semi-distributed', lumped parameter models without prior calibration against a physically based model and the potential for implying unexpected and possibly implausible hydrological characteristics through the specification of flows without considering whether they could occur for realistic soil depths and properties. As there is a need for application of such SDLP models, particularly when undertaking probabilistic calculations, it is suggested that, in future, explicit hydrological modelling should be undertaken first, so that a physically realistic representation can be produced as a basis for assessment studies of the migration of radionuclides or other contaminants. PMID- 15798278 TI - Uncertainty and power at low levels of incurred radiation dose. AB - It is common practice when calculating dose to exposed populations to average the variables that go into the dose calculation (e.g. environmental concentrations, air kerma, consumption rates, occupancy rates). This approach is simple and can be useful where data are obtained over different periods (weekly, monthly, quarterly), where samples may be bulked for some analyses but not others and where gaps in the data are present. However, such an approach does not yield information on the degree of uncertainty around the average dose calculated. An alternative approach is to estimate the dose to each individual and to obtain an average from this data set, which can then also be used to derive a measure of uncertainty around the central dose estimate. In this study, we demonstrate the variability in dose estimates using a hypothetical data set and consider the implications for sample size to achieve fixed confidence or resolving power. We recommend calculating the dose to every individual sampled, in order both to obtain the average dose and to estimate its variability. We argue that it is best practice to obtain information as complete as possible from the available sample of individuals. PMID- 15798279 TI - Field and model investigations of external gamma dose rates along the Cumbrian coast, NW England. AB - A survey of the contribution to external dose from gamma rays originating from intertidal sediments in the vicinity of the British Nuclear Group Sellafield site showed that the major anthropogenic contributions were due to (137)Cs and (60)Co. At some sites, traces of other anthropogenic radionuclides were detected, namely (106)Ru, (125)Sb, and (154)Eu. The proportions of fine grained material (<63 microm) were used to improve model predictions of dose contribution due to external exposure to gamma rays, using the CUMBRIA77/DOSE77 model. Model dose predictions were compared to those directly measured in the field. Using the new proportions of fine grained material (1-17.5%) in conjunction with field gamma ray spectra, model predictions were improved considerably for most sites. Exceptions were at Drigg Barn Scar and Whitehaven Coal Sands sites, which had their own unique characteristics. The highest (60)Co activity concentrations in this study were detected at Drigg Barn Scar. These relatively high activity concentrations of (60)Co were due to the presence of (60)Co in mussels and barnacles, hence upsetting the fine sediment relationships used in previous dose calculations. Whitehaven Coal Sands was unusual in that it contained higher levels of radionuclides than would be expected in sandy sediment. The mineralogy of these sediments was the controlling factor on (137)Cs binding, rather than the proportion of fine grained material. By adjusting the effective fine grained sediment proportions for calculations involving (60)Co and (137)Cs at Drigg Barn Scar and Whitehaven Coal Sands respectively, the CUMBRIA77/DOSE77 model predictions could be improved upon significantly for these sites. This work highlights the influence of particle size and sediment composition on external dose rate calculations, as well as the potential for external dose contributions from biota. PMID- 15798280 TI - Chromosome intra- and inter-changes determined by G-banding in radiation workers with in vivo exposure to plutonium. AB - Suggestions that exposure to intakes of alpha-emitting radionuclides such as plutonium could result in a specific profile of chromosome damage distinguishable from that of low LET irradiation have led to the re-analysis of the different types of chromosome aberrations in peripheral blood lymphocytes determined by G banding in a group of 20 plutonium workers from the British Nuclear Fuels plc facility at Sellafield, UK. Comparisons were made with a group of workers with negligible plutonium intakes but similar external gamma doses and with an unexposed control group. Examination of simple translocation frequencies in the three groups indicated a significant difference (P = 0.033), with the higher frequency in the plutonium workers indicating that exposure from plutonium was contributing to the aberration yield. Slightly raised frequencies of both intra chromosomal and complex aberrations were observed in the plutonium workers in comparison with the comparable external exposure group and the control group but the difference did not reach significance at the P = 0.05 level and there was no variation in the relative frequencies of the different aberration types between the three groups. There was, therefore, no firm indication from this study that either intra-chromosomal or complex aberrations could be used as a specific marker of high LET exposure in workers with historical intakes of plutonium. PMID- 15798282 TI - The influence of gamma radiation on the electro-optical characteristics of CR-39 polymer. AB - The influence of gamma radiation on the electro-optical properties of CR-39 polymer has been investigated by capacitive and interferometric measurements to see the order and tendency of irradiation effects on polymer CR-39, which is commonly used in track detectors. Seven samples of thickness 760 microm and other dimensions 25 mm x 25 mm have been exposed to gamma radiation in the range of zero to 70 kGy in 10 kGy steps. Experimental results exhibit radiation-dependent degradation even at relatively low doses. Modulation of electronic properties can be exploited as an alternative diagnosis tool. PMID- 15798281 TI - Analysis of the RB1 gene in children with retinoblastoma having residential connections to West Cumbria, England. AB - Six of eight cases of retinoblastoma previously identified as having a residential association with West Cumbria, England, in which the Sellafield nuclear installation is situated, were examined for the presence of a constitutional RB1 mutation. No mutations were detected, thus providing strong evidence against an environmental or occupational genotoxic effect causing germline mutations in the parents of these children. PMID- 15798283 TI - Radiation quantities and units--understanding the sievert. AB - International recommendations for changes to the SI brochure regarding the definition and use of the quantity dose equivalent are presented. The second part of this note is a simple explanation of the difference between a radiation quantity and its unit. PMID- 15798287 TI - SRP South West Regional Conference: codes and methods and prior risk assessments. PMID- 15798284 TI - Concerning CERRIE's conclusions and COMARE's response on doses and risks from internal emitters. PMID- 15798289 TI - System identification: a multi-signal approach for probing neural cardiovascular regulation. AB - Short-term, beat-to-beat cardiovascular variability reflects the dynamic interplay between ongoing perturbations to the circulation and the compensatory response of neurally mediated regulatory mechanisms. This physiologic information may be deciphered from the subtle, beat-to-beat variations by using digital signal processing techniques. While single signal analysis techniques (e.g., power spectral analysis) may be employed to quantify the variability itself, the multi-signal approach of system identification permits the dynamic characterization of the neural regulatory mechanisms responsible for coupling the variability between signals. In this review, we provide an overview of applications of system identification to beat-to-beat variability for the quantitative characterization of cardiovascular regulatory mechanisms. After briefly summarizing the history of the field and basic principles, we take a didactic approach to describe the practice of system identification in the context of probing neural cardiovascular regulation. We then review studies in the literature over the past two decades that have applied system identification for characterizing the dynamical properties of the sinoatrial node, respiratory sinus arrhythmia, and the baroreflex control of sympathetic nerve activity, heart rate and total peripheral resistance. Based on this literature review, we conclude by advocating specific methods of practice and that future research should focus on nonlinear and time-varying behaviors, validation of identification methods, and less understood neural regulatory mechanisms. Ultimately, we hope that this review stimulates such future investigations by both new and experienced system identification researchers. PMID- 15798290 TI - Microfluidics for flow cytometric analysis of cells and particles. AB - This review describes recent developments in microfabricated flow cytometers and related microfluidic devices that can detect, analyze, and sort cells or particles. The high-speed analytical capabilities of flow cytometry depend on the cooperative use of microfluidics, optics and electronics. Along with the improvement of other components, replacement of conventional glass capillary based fluidics with microfluidic sample handling systems operating in microfabricated structures enables volume- and power-efficient, inexpensive and flexible analysis of particulate samples. In this review, we present various efforts that take advantage of novel microscale flow phenomena and microfabrication techniques to build microfluidic cell analysis systems. PMID- 15798291 TI - Flow dependence and repeatability of interrupter resistance in lower airways and nose. AB - The interrupter technique, the simplest method for measuring airflow resistance (R(int)) is particularly valuable under field conditions. We investigate whether during tidal breathing, variations in the flow at which interruption occurs contribute to variability of results. Using a portable device with mouthpiece, sets of 10 measurements of R(int) (R(int,mo)) were made in inspiration and expiration at 0.05 l s(-1) intervals from 0.1 up to 0.9 l s(-1) flow in 22 normal adults, 11 children (5-9 years) and 12 COPD patients. R(int) was also measured via nasal-mask in normal adults (R(int,na)). Intra-subject coefficient of variation was obtained at each flow and flow-dependence of R(int) was assessed. In normal subjects, R(int)-flow relationships were consistent, with a narrow range of absolute values. R(int,na), but not R(int,mo), rose with increasing flow, especially >0.4 l s(-1). Repeatability was poor at flows <0.3 l s(-1) but improved with increasing flow and was better in inspiration than expiration. In children, repeatability was better than in adults and R(int,mo) was not flow dependent at < or =0.4 l s(-1). By contrast, in COPD patients repeatability was less good and R(int,mo) increased with increasing flows. R(int,mo) and R(int,na) should be measured at fixed inspiratory flows. The best signal-to-noise ratios were obtained at 0.4 l s(-1) for R(int) in normal adults and COPD patients and at 0.3 l s(-1) in children. PMID- 15798292 TI - Modeling for radio-frequency conductive keratoplasty: implications for the maximum temperature reached in the cornea. AB - Conductive keratoplasty (CK) is a new surgical technique for steepening the contours of the cornea to reduce hyperopia. It has been emphasized that during CK, tissue resistance to radio-frequency electrical current flow generates a localized heat with temperatures between 65 and 75 degrees C; however, we hypothesize that the maximum temperature reached in the cornea may be higher. For this reason, we developed a finite-element model to estimate the temperature distributions in the cornea during CK. The time evolution of the impedance obtained from computer simulations was compared to that obtained in an experimental study previously published. Our results show that during a typical CK with a 60% setting power (equivalent to 200 V peak-to-peak), the cornea may reach temperatures over 100 degrees C at the electrode tip. On the other hand, the initial impedance of the cornea has a significant influence on the temperature distribution, while the initial temperature of the cornea is not a significant parameter. The results also suggest that low power settings (30-40%) do not produce temperatures over 100 degrees C. Finally, although the actual voltage waveform during CK is exponential and pulsed, our model based on a constant voltage (with a value equal to the root mean square value) provides a better agreement between the theoretical impedance time evolution and that obtained experimentally. PMID- 15798293 TI - A comparative study of different references for EEG spectral mapping: the issue of the neutral reference and the use of the infinity reference. AB - Based on EEG data recorded from 11 subjects with eyes open and the left mastoid (M) reference, three data sets were generated by re-referencing to the conventional linked mastoids (L), average (A) and the new 'infinity' (I) reference provided by the reference electrode standardization technique (REST, Yao 2001 Physiol. Meas. 22 693-711). The EEG power in the alpha frequency band with the four different references was calculated and compared with respect to the total energy and spatial amplitude weight centre (AWC) coordinates, to compare the effects of different references on power mapping in the frequency domain. Compared with the I reference, the AWCs of the EEG with the M reference show significant shifts to the right, frontal and superficial positions, the L reference significant shifts to frontal and superficial positions, and the A reference shifts the AWC significantly to a deeper position. Furthermore, the power maps of the M and L references have larger total power than the I reference, while that of the A reference has the smallest total power. These results confirm that different choices of reference electrodes result in systematic changes in the distribution of EEG frequency power, and in order to reduce the effect of such systematic shifts on the explanation of EEG mappings, a common reference is necessary for EEG research. We recommend the I reference for objective use in cross-laboratory studies and clinical practices, as it is far from all the other electrodes and can act as a neutral reference. PMID- 15798295 TI - Simultaneous monitoring of separate fetal magnetocardiographic signals in twin pregnancy. AB - Fetal magnetocardiography (fMCG) allows the non-invasive recording of fetal cardiac electrical activity with increasing efficacy as gestation progresses. Many reports on the successful extraction of reliable fetal magnetocardiographic traces in singleton pregnancies exist in the literature, whereas there is only one report on the reconstruction of averaged fetal cardiac signals obtained in a twin pregnancy with the use of a double sensor array system. In this paper, we aimed at assessing the effectiveness of an ICA-based procedure to reconstruct the time course of fetal cardiac signals recorded with a single-shot multi-channel fMCG device in an uncomplicated twin pregnancy at 27 weeks. The evaluation of heart rate and beats synchronicity permitted the differentiation of fetal components; the quality of reconstructed fetal signals allowed visual inspection on single cycles and the simultaneous monitoring of separate fetal heart rate patterns. The proposed technique might be applied in twin pregnancies not only to characterize fetal arrhythmias, but also in all cases of discordant fetal growth, either in the case of intra-uterine growth retardation affecting one fetus, or in the case of twin-twin transfusion syndrome, a life-threatening condition where both fetuses are at risk of heart failure. PMID- 15798294 TI - Monitoring of lung edema using focused impedance spectroscopy: a feasibility study. AB - Currently only ionizing or invasive methods are used in clinical applications for the monitoring of extracellular lung water. Alternatively a method called focused conductivity spectroscopy (FCS) is suggested, which aims at reconstructing a pulmonary edema index (PEIX) by measuring the electrical conductivity of the region of interest (ROI) at several frequencies. In contrast to electrical impedance tomography (EIT) a minimum number of strategically placed electrodes is used. The goals of this study were the analysis of the sensitivity for the PEIX, an estimate of the optimal electrode configuration and the determination of the required frequencies. In order to calculate the solution of the FCS forward problem a realistic 3D model of a human torso was developed containing both lungs, the heart, the liver and the thorax musculature. The bioelectrical properties for each compartment were described with appropriate tissue models which relate the conductivity spectra to physiological parameters. The PEIX was defined as the interstitial volume fraction of the alveolar septa. Furthermore the model includes 48 electrodes subdivided into three layers. The optimal electrode configuration was selected by minimizing the number of electrodes, among certain subsets of these electrodes. The analysis shows that eight to ten electrodes and six frequencies are theoretically sufficient to obtain a coefficient of variation. PMID- 15798296 TI - Simultaneous measurement of rat brain cortex PtO2 using EPR oximetry and a fluorescence fiber-optic sensor during normoxia and hyperoxia. AB - Electron paramagnetic resonance (EPR) oximetry is a promising, relatively non invasive method of monitoring tissue partial pressure of oxygen (PtO(2)) that has proven useful in following changes in PtO(2) under various physiologic and pathophysiologic conditions. Optimal utilization of the method will be facilitated by systematic comparisons with other available methods. Here, we report on the absolute values and changes of rat brain PtO(2) using EPR oximetry and the OxyLite, an oxygen monitor based on fluorescence quenching, at adjacent locations in the same brain. EPR oximetry utilizes an implanted oxygen-sensitive material and reports tissue PtO(2) at the surface of the material. OxyLite measures PtO(2) using the fluorescence lifetime of a chromophore fixed to the tip of an optical fiber that is inserted into tissue. Measurements were made at a depth of 2-3 mm into the cortex during normoxia and during breathing of carbogen (95% O(2):5% CO(2)) followed by a return to normoxia. We conclude that in this study (1) PtO(2) values reported by the two methods are similar but not exactly the same, (2) both methods can record a baseline and rapid changes in PtO(2), (3) changes in PtO(2) induced by increasing FiO(2) from 0.26 to 0.95 (carbogen) were similar by the two methods and (4) in some rats breathing carbogen, absolute values of PtO(2) were above the sensitive range of the OxyLite method. PMID- 15798297 TI - Feasibility of the use of a novel soft tissue stiffness meter. AB - Simple but objective measurement of soft tissue consistency would be advantageous in the assessment of many neurological, lymphostatic and venous disorders. The aim of the present study was to evaluate the feasibility of using a novel hand held computerized soft tissue stiffness meter (STSM). The STSM describes the soft tissue stiffness (STS) in the form of the instantaneous force (N) by which the tissue resists the constant deformation produced by a cylindrical intender. Firstly, the STSM was used to test elastomer samples with known mechanical properties. In the in vivo assessment, 12 healthy, nondisabled adults (age range, 24-57 years) and 16 subjects with chronic myofascial neck pain syndrome (age range, 27-55 years) were studied. To study the reproducibility (coefficient of variation (CV(%))) of the method, the measurement sites were either marked with a marker pen (marked points) or localized anatomically (unmarked points). Measurements were made from the dorsal forearm (Arm), trapezius (Tra), levator scapulae (Lev), infraspinatus (Inf) and deltoideus (Del) muscle areas. STS in the forearm was studied during different types of short-term relative isometric loading of the muscle as well as during venous occlusion. STS values of the myofascial trigger points in the Lev muscles were evaluated bilaterally. A linear, positive relationship was found between the indenter force (N) and the dynamic compressive modulus (MPa) of elastomer stiffness (r(2) = 0.90, n = 9). Intra- and interrater CVs of marked and unmarked sites varied between 4.31% and 12.06%. STS increased linearly along the relative muscle load (r(2) = 0.96) and nonlinearly during the venous occlusion (r(2) = 0.97). Statistically significant regional variation of STS was found between the different measurement sites (p < 0.05). In conclusion, STSM can evaluate tissue stiffness quantitatively and yield reproducible data. PMID- 15798298 TI - Effect of esophagus status and catheter configuration on multiple intraluminal impedance measurements. AB - Multiple intraluminal impedance measurement is used to investigate the esophagus condition. This procedure provides information about the esophagus status, reflux occurrence and clearance mechanism. This paper presents finite-element approximation of the forward problem of the catheter inside the esophagus, relating the real data obtained from multiple intraluminal impedance to the solution obtained from the finite-element approximation. Investigation of the effect of the esophagus status on the impedance, and the correlation between various factors that affect the impedance are presented. The results of this paper provide theoretical bases for relating the esophagus condition to the impedance waveforms. PMID- 15798299 TI - Inertance measurements by jet pulses in ventilated small lungs after perfluorochemical liquid (PFC) applications. AB - Perfluorochemical liquid (PFC) liquids or aerosols are used for assisted ventilation, drug delivery, lung cancer hyperthermia and pulmonary imaging. The aim of this study was to investigate the effect of PFC liquid on the inertance (I) of the respiratory system in newborn piglets using partial liquid ventilation (PLV) with different volumes of liquid. End-inspiratory (I(in)) and end expiratory (I(ex)) inertance were measured in 15 ventilated newborn piglets (age < 12 h, mean weight 724 +/- 93 g) by brief flow pulses before and 80 min after PLV using a PFC volume (PF5080, 3 M) of 10 ml kg(-1) (N = 5) or 30 ml kg(-1) (N = 10). I was calculated from the imaginary part of the measured respiratory input impedance by regression analysis. Straight tubes with 2-4 mm inner diameter were used to validate the equipment in vitro by comparison with the analytically calculated values. In vitro measurements showed that the measuring error of I was <5% and that the reproducibility was better than 1.5%. The correlation coefficient of the regression model to determine I was >0.988 in all piglets. During gas ventilation, I(in) and I(ex) (mean +/- SD) were 31.7 +/- 0.8 Pa l(-1) s(2) and 33.3 +/- 2.1 Pa l(-1) s(2) in the 10 ml group and 32.4 +/- 0.8 Pa l(-1) s(2) and 34.0 +/- 2.5 Pa l(-1) s(2) in the 30 ml group. However, I of the 3 mm endotracheal tube (ETT) used was already 26.4 Pa l(-1) s(2) (about 80% of measured I). During PLV, there was a minimal increase of I(in) to 33.1 +/- 2.5 Pa l(-1) s(2) in the 10 ml group and to 34.5 +/- 2.7 Pa l(-1) s(2) in the 30 ml group. In contrast, the increase of I(ex) was dramatically larger (p < 0.001) to 67.7 +/- 13.3 Pa l(-1) s(2) and to 74.8 +/- 9.3 Pa l(-1) s(2) in the 10 ml and 30 ml groups, respectively. Measurements of I by jet pulses in intubated small animals are reproducible. PFC increases the respiratory inertance, but the magnitude depends considerably on its spatial distribution which changes during the breathing cycle. Large differences between I(in) and I(ex) are an indicator for liquid in airways or the ETT. PMID- 15798300 TI - Modelling the electrical properties of bladder tissue--quantifying impedance changes due to inflammation and oedema. AB - Electrical impedance spectroscopy has been developed as a potential method for the diagnosis of carcinoma in epithelial tissues. An understanding of the influence of structural changes in the tissue on the properties measured using this technique is essential for interpreting measured data and optimization of probe design. In contrast to other tissue types, carcinoma in situ of the bladder gives rise to an increase in electrical impedance over the kHz-MHz frequency range in comparison to normal tissue. Finite element models of the urothelium and the underlying superficial lamina propria have been constructed and solved in order to ascertain the influence of structural changes associated with malignancy, oedema and inflammation on the measured electrical properties of the tissue. Sensitivity analysis of results from a composite tissue model suggests that the increase in lymphocyte density in the lamina propria associated with an inflammatory response to the infiltration of urine into the tissue may explain these unusual electrical properties. PMID- 15798301 TI - Pressure reflectometry: in vitro recordings with a new technique for simultaneous measurement of cross-sectional area and pressure in a collapsible tube. AB - Pressure reflectometry is a new and easily applicable technique for simultaneous measurement of cross-sectional area and pressure in a collapsible biological tube. Related values of pressure and cross-sectional area are relevant for assessment of the mechanical properties of a biological tube. A very thin and highly flexible 6 cm long polyurethane bag with a diameter of 5 mm when expanded was introduced into test models. The cross-sectional area of the first 6 cm of the cavities up to 16 mm(2) (depending of the size of the bag) can be measured at the same time. The cross-sectional area was measured by acoustic reflectometry while pressures of 10-200 cmH(2)O were applied by a pump. The echoes from the cavity were analyzed and displayed by a computer technique. The aim of the study was to evaluate the technical capabilities and limitations of this new method. The accuracy and reproducibility were examined in eight different models with known cross-sectional areas in the interval of 4-16 mm(2). Two of the models had two successive constrictions reducing the cavity to 50% and 25% respectively. Measurements were reliable from about 1 to 5 cm within the cavity and at pressures from 10 to 200 cmH(2)O. The absolute error in models without constrictions did not exceed 0.9 mm(2) and at the first constriction the error did not exceed 1.2 mm(2). In cavities without constrictions, the coefficients of variation (CVs) were 0.2-11%. In models with constrictions the CVs were 6-25% at the first constriction when different pressures were applied to the plastic bag. The reproducibility was not influenced significantly by change in background noise, temperature, catheters, geometries of the cavities or new calibration. In conclusion, the accuracy of this new technique depends on the size of the measured cavity, on the pressure in the models and whether the model has constrictions or not. The measurements seem reliable for clinical use in the range of 4-16 mm(2) and at pressures from 10 to 200 cmH(2)O. PMID- 15798302 TI - Compliance and resistance levels and unloading in proportional assist ventilation. AB - During proportional assist ventilation (PAV), resistive and elastic unloading relieve the work of breathing. Excessive unloading, however, results in resonant oscillations and runaway pressures. Our aim was to determine the appropriate levels of unloading that could be applied to clinical practice. A lung model, resistance (50 or 150 cmH(2)O l(-1) s(-1)) and compliance (0.4 or 0.8 ml/cmH(2)O), was used. The volumes and airway pressures delivered by the ventilator at varying levels of resistive and elastic unloading and simulated breaths were recorded. Oscillations in airway pressure only occurred when the level of resistive unloading exceeded the model's resistance. When the level of unloading fully compensated for the model's compliance, peak inflating pressures greater than 40 cmH(2)O were delivered; peak pressure limits of 20 cmH(2)O, however, resulted in very short (0.2 s or less) inflation times. High peak pressures were not delivered if the level of elastic unloading used was limited to that which reduced the model's elastance to that of a 'normal lung'. In conclusion, these results suggest that when using PAV, it is important to assess the compliance and resistance of the infant and endotracheal tube, so that levels of unloading that fully compensate for the resistance and compliance levels can be avoided. PMID- 15798303 TI - Evaluation of arterial endothelial function using transit times of artificially induced pulses. AB - Impairment of arterial endothelial function is an early event in atherosclerosis and correlates with the major risk factors for cardiovascular disease. The most widely employed non-invasive measure of endothelial function involves brachial artery (BA) diameter measurement using ultrasound imaging before and after several minutes of blood flow occlusion. The change in arterial diameter is a measure of flow-mediated vasorelaxation (FMVR). The high between-laboratory variability of results and cost of instrumentation render this technique unsuitable for routine clinical use. We induce artificial pulses at the superficial radial artery using a linear actuator. An ultrasonic Doppler stethoscope detects these pulses 10-30 cm proximal to the point of pulse induction. The delay between pulse application and detection provides the pulse transit time (PTT). By measuring PTT before and after 5 min of BA occlusion and ensuing reactive hyperemia, FMVR may be measured based on the changes in PTT caused by changes in vessel caliber, smooth muscle tone and wall thickness. We (1) validate the sensitivity of this technique to arterial wall tone using sublingual nitroglycerin and (2) compare measurements of endothelial function to ultrasound BA diameter measurements in 12 human subjects. The PTT-based method is verified to measure arterial wall tone and is shown to provide 37% greater sensitivity (p < 0.05) to FMVR than BA diameter measurements. By measuring the change in pulse transit time before and after endothelial stimulus, a sensitive, reproducible and convenient measure of endothelial function may be obtained at a low cost. PMID- 15798304 TI - Non-invasive measurement of intrapulmonary shunt during inert gas rebreathing. AB - We tested the agreement between non-invasive measurement of intrapulmonary shunt, using oxygen uptake and pulmonary capillary blood flow measurement obtained by nitrous oxide rebreathing, with that measured using mixed venous blood sampling. Nine patients were recruited pre- and post-cardiac surgery resulting in 20 sets of measurements overall. Mean shunt fraction was 12.5%, and bias between methods (+/-95% confidence limits) was -0.7% (+/-0.8%). The standard deviation of the difference was 1.7% with limits of agreement between the two methods of +2.6% and -3.9%. Correlation coefficient r was 0.90. Agreement with the invasive standard was less accurate and precise where cardiac output was measured by bolus thermodilution (mean bias +1.6%, standard deviation of the difference 2.2%, limits of agreement between the two methods of +5.8% and -2.8%, r = 0.86). Good agreement was demonstrated between the non-invasive method and the invasive reference standard. PMID- 15798305 TI - Predictive regression equations and clinical uses of peripheral pulse timing characteristics in children. AB - Studies have shown that increased arterial stiffening can be an indication of cardiovascular diseases like hypertension. In clinical practice, this can be detected by measuring the blood pressure (BP) using a sphygmomanometer but it cannot be used for prolonged monitoring. It has been established that pulse wave velocity (PWV) is a direct measure of arterial stiffening but its usefulness is hampered by the absence of non-invasive techniques to estimate it. Pulse transit time (PTT) is a simple and non-invasive method derived from PWV. However, limited knowledge of PTT in children is found in the present literature. The aims of this study are to identify independent variables that confound PTT measure and describe PTT regression equations for healthy children. Therefore, PTT reference values are formulated for future pathological studies. Fifty-five Caucasian children (39 male) aged 8.4 +/- 2.3 yr (range 5-12 yr) were recruited. Predictive equations for PTT were obtained by multiple regressions with age, vascular path length, BP indexes and heart rate. These derived equations were compared in their PWV equivalent against two previously reported equations and significant agreement was obtained (p < 0.05). Findings herein also suggested that PTT can be useful as a continuous surrogate BP monitor in children. PMID- 15798306 TI - An in vitro assessment of gas trapping during high frequency oscillation. AB - Our aim was to assess the influence of oscillatory frequency, inspiratory-to expiratory (I:E) ratio and airway pressure on gas trapping during high frequency oscillatory ventilation (HFOV). A lung model was used, which had a high compliance (4.0 ml (cmH(2)O)(-1)) and resistance (160 cmH(2)O l(-1) s(-1)) resulting in a long time constant (0.64 s). To assess whether gas trapping occurred, the mean pressure within the lung model (equivalent to alveolar pressure) was compared to the mean airway pressure (MAP) measured at the manifold (manifold MAP) of the two oscillators used, the SLE 5000 (I:E ratio 1:1) and the Sensor Medics 3100A (tested at I:E ratios of 1:2 and 1:1). The effects were assessed of raising the MAP from 15 to 35 cmH(2)O, the oscillatory amplitude from 30 to 70 cmH(2)O and the frequency from 5 to 15 Hz (5 to 20 Hz SLE only). There were no significant trends for differences between the pressure within the lung model and the 'manifold' MAP to increase (i.e. no evidence of gas trapping), as MAP, amplitude or frequency was increased, regardless of which oscillator or I:E ratio was used. Increasing the pressure amplitude led to a progressive fall in the pressure within the lung model when an I:E ratio of 1:2 was used (p < 0.05). Our results suggest that significant gas trapping does not occur during HFOV even if there is high compliance and resistance. PMID- 15798307 TI - Prediction of radiation-induced normal tissue complications in radiotherapy using functional image data. AB - The aim of this study has been to explicitly include the functional heterogeneity of an organ as a factor that contributes to the probability of complication of normal tissues following radiotherapy. Situations for which the inclusion of this information can be advantageous to the design of treatment plans are then investigated. A Java program has been implemented for this purpose. This makes use of a voxelated model of a patient, which is based on registered anatomical and functional data in order to enable functional voxel weighting. Using this model, the functional dose-volume histogram (fDVH) and the functional normal tissue complication probability (fNTCP) are then introduced as extensions to the conventional dose-volume histogram (DVH) and normal tissue complication probability (NTCP). In the presence of functional heterogeneity, these tools are physically more meaningful for plan evaluation than the traditional indices, as they incorporate additional information and are anticipated to show a better correlation with outcome. New parameters m(f), n(f) and TD(50f) are required to replace the m, n and TD(50) parameters. A range of plausible values was investigated, awaiting fitting of these new parameters to patient outcomes where functional data have been measured. As an example, the model is applied to two lung datasets utilizing accurately registered computed tomography (CT) and single photon emission computed tomography (SPECT) perfusion scans. Assuming a linear perfusion-function relationship, the biological index mean perfusion weighted lung dose (MPWLD) has been extracted from integration over outlined regions of interest. In agreement with the MPWLD ranking, the fNTCP predictions reveal that incorporation of functional imaging in radiotherapy treatment planning is most beneficial for organs with a large volume effect and large focal areas of dysfunction. There is, however, no additional advantage in cases presenting with homogeneous function. Although presented for lung radiotherapy, this model is general. It can also be applied to positron emission tomography (PET)-CT or functional magnetic resonance imaging (fMRI)-CT registered data and extended to the functional description of tumour control probability. PMID- 15798308 TI - Development of the female voxel phantom, NAOMI, and its application to calculations of induced current densities and electric fields from applied low frequency magnetic and electric fields. AB - This paper outlines the development of a 2 mm resolution voxel model, NAOMI (aNAtOMIcal model), designed to be representative of the average adult female. The primary medical imaging data were derived from a high-resolution MRI scan of a 1.65 m tall, 23 year old female subject with a mass of 58 kg. The model was rescaled to a height of 1.63 m and a mass of 60 kg, the dimensions of the International Commission on Radiological Protection reference adult female. There are 41 tissue types in the model. The application of NAOMI to the calculations of induced current densities and electric fields from applied low frequency magnetic and electric fields is described. Comparisons are made with values from the male voxel model, NORMAN. The calculations were extended from 50 Hz up to 10 MHz. External field reference levels are compared with the ICNIRP guidelines. PMID- 15798309 TI - Modelling and validation of spectral reflectance for the colon. AB - The spectral reflectance of the colon is known to be affected by malignant and pre-malignant changes in the tissue. As part of long-term research on the derivation of diagnostically important parameters characterizing colon histology, we have investigated the effects of the normal histological variability on the remitted spectra. This paper presents a detailed optical model of the normal colon comprising mucosa, submucosa and the smooth muscle layer. Each layer is characterized by five variable histological parameters: the volume fraction of blood, the haemoglobin saturation, the size of the scattering particles, including collagen, the volume fraction of the scattering particles and the layer thickness, and three optical parameters: the anisotropy factor, the refractive index of the medium and the refractive index of the scattering particles. The paper specifies the parameter ranges corresponding to normal colon tissue, including some previously unpublished ones. Diffuse reflectance spectra were modelled using the Monte Carlo method. Validation of the model-generated spectra against measured spectra demonstrated that good correspondence was achieved between the two. The analysis of the effect of the individual histological parameters on the behaviour of the spectra has shown that the spectral variability originates mainly from changes in the mucosa. However, the submucosa and the muscle layer must be included in the model as they have a significant constant effect on the spectral reflectance above 600 nm. The nature of variations in the spectra also suggests that it may be possible to carry out model inversion and to recover parameters characterizing the colon from multi spectral images. A preliminary study, in which the mucosal blood and collagen parameters were modified to reflect histopathological changes associated with colon cancer, has shown that the spectra predicted by our model resemble measured spectral reflectance of adenocarcinomas. This suggests that an extended model, which incorporates parameters corresponding to an abnormal colon, may be effective for differentiation between normal and cancerous tissues. PMID- 15798310 TI - A comparative EPR, infrared and Raman study of natural and deproteinated tooth enamel and dentin. AB - The aim of the present work was to investigate how the native signal observed in the electron paramagnetic resonance (EPR) spectrum of tooth enamel and dentin is associated with the organic content of the two tissues. This was achieved by comparing the EPR native signal and the optical bands (Raman and infrared, IR) associated with organic components of tooth enamel and dentin, in natural and deproteinated samples. The main results were: (a) in natural samples, the organic optical bands are more intense in dentin than in enamel, in contrast with the EPR native signal which shows similar intensity in the two tissues; (b) after deproteination, the optical organic bands are completely suppressed in both dentin and enamel, while the EPR native signal is eliminated only in dentin. It is suggested that the IR and Raman organic bands are originated in the bulk of the organic matrix, while the paramagnetic centres associated with the EPR native signal are located in the organic-mineral interface. PMID- 15798311 TI - Ir-192 HDR transit dose and radial dose function determination using alanine/EPR dosimetry. AB - Source positioning close to the tumour in high dose rate (HDR) brachytherapy is not instantaneous. An increment of dose will be delivered during the movement of the source in the trajectory to its static position. This increment is the transit dose, often not taken into account in brachytherapeutic treatment planning. The transit dose depends on the prescribed dose, number of treatment fractions, velocity and activity of the source. Combining all these factors, the transit dose can be 5% higher than the prescribed absorbed dose value (Sang-Hyun and Muller-Runkel, 1994 Phys. Med. Biol. 39 1181-8, Nath et al 1995 Med. Phys. 22 209-34). However, it cannot exceed this percentage (Nath et al 1995). In this work, we use the alanine-EPR (electron paramagnetic resonance) dosimetric system using analysis of the first derivative of the signal. The transit dose was evaluated for an HDR system and is consistent with that already presented for TLD dosimeters (Bastin et al 1993 Int. J. Radiat. Oncol. Biol. Phys. 26 695-702). Also using the same dosimetric system, the radial dose function, used to evaluate the geometric dose degradation around the source, was determined and its behaviour agrees better with those obtained by Monte Carlo simulations (Nath et al 1995, Williamson and Nath 1991 Med. Phys. 18 434-48, Ballester et al 1997 Med. Phys. 24 1221-8, Ballester et al 2001 Phys. Med. Biol. 46 N79-90) than with TLD measurements (Nath et al 1990 Med. Phys. 17 1032-40). PMID- 15798314 TI - Reducing the number of monitor units in multileaf collimator field segmentation. AB - Multileaf collimators (MLCs) are the prevailing tool for the realization of radiation fields in intensity modulated radiation therapy (IMRT). One step in the treatment planning is to determine a set of leaf positions realizing a certain intensity modulated radiation field. In this paper we suggest two extensions in the use of the MLC that lead to considerable savings in terms of monitor units, thus potentially increasing the treatment quality. We test our method with random and with clinical sample matrices. PMID- 15798313 TI - Quantitative comparison of suitability of various beams for range monitoring with induced beta+ activity in hadron therapy. AB - In radiation therapy with hadron beams, it is important to evaluate the range of incident ions and the deposited dose distribution in a patient body for the effective utilization of such properties as the dose concentration and the biological effect around the Bragg peak. However, there is some ambiguity in determining this range because of a conversion error from the x-ray CT number to the charged particle range. This is because the CT number is related to x-ray absorption coefficients, while the ion range is determined by the electron density of the substance. Using positron emitters produced in the patient body through fragmentation reactions during the irradiation has been proposed to overcome this problem. The activity distribution in the patient body can be deduced by detecting pairs of annihilation gamma rays emitted from the positron emitters, and information about the range of incident ions can be obtained. In this paper, we propose a quantitative comparison method to evaluate the mean range of incident ions and monitor the activity distribution related to the deposited dose distribution. The effectiveness of the method was demonstrated by evaluating the range of incident ions using the maximum likelihood estimation (MLE) method and Fisher's information was calculated under realistic conditions for irradiations with several kinds of ions. From the calculated Fisher's information, we compared the relative advantages of initial beams to determine the range of incident ions. The (16)O irradiation gave the most information among the stable heavy ions when we measured the induced activity for 500 s and 60 s just after the irradiation. Therefore, under these conditions, we concluded that the (16)O beam was the optimum beam to monitor the activity distribution and to evaluate the range. On the other hand, if the positron emitters were injected directly as a therapeutic beam, the (15)O irradiation gave the most information. Although the relative advantages of initial beams as well as the measured activity distributions slightly varied according to the measurement conditions, comparisons could be made for different conditions by using Fisher's information. PMID- 15798312 TI - An experimental and Monte Carlo investigation of the energy dependence of alanine/EPR dosimetry: II. Clinical electron beams. AB - The energy dependence of alanine/EPR dosimetry for 8, 12, 18 and 22 MeV clinical electron beams was investigated by experiment and by Monte Carlo simulations. Alanine pellets in a waterproof holder were irradiated in a water phantom using an Elekta Precise linear accelerator. The dose rates at the reference point were determined following the TG-51 protocol using an NACP-02 parallel-plate chamber calibrated in a (60)Co beam. The EPR spectra of irradiated pellets were measured using a Bruker EMX 081 EPR spectrometer. Experimentally, we found no significant change in alanine/EPR response to absorbed dose-to-water over the energy range 8 22 MeV at an uncertainty level of 0.6%. However, the response for high-energy electrons is about 1.3 (+/-1.1)% lower than for (60)Co. The EGSnrc Monte Carlo system was used to calculate the ratio of absorbed dose-to-alanine to absorbed dose-to-water and it was shown that there is 1.3 (+/-0.2)% reduction in this ratio from the (60)Co beam to the electron beams, which confirms the experimental results. Alanine/EPR response per unit absorbed dose-to-alanine was also investigated and it is the same for high-energy electrons and (60)Co gamma-rays. PMID- 15798315 TI - The effect on IMRT conformality of elastic tissue movement and a practical suggestion for movement compensation via the modified dynamic multileaf collimator (dMLC) technique. AB - A major remaining problem in delivering radiotherapy, specifically intensity modulated radiation therapy (IMRT), is the need to accommodate and correct for intrafraction movement. The developing availability of 4D computed tomographic images can potentially form the basis of the new field of image-guided IMRT. It is important to understand the effects on delivered dose of the patient breathing during IMRT and this paper models the effect which applies whether there is or is not a time component to the IMRT delivery method. It then goes on to suggest a practical correction strategy. The 'stretch-and-shift-the-planned-modulations' strategy is proposed and a practical method to deliver this is explained. This practical strategy is based on a modification of the dynamic multileaf collimator IMRT method whereby the leaves are arranged to 'breath' in tandem with the breathing of the patient. Some examples are also given from a study of mismatching the patient and leaf-correction motions. PMID- 15798316 TI - Effects on electron beam penumbra using the photon MLC to reduce bremsstrahlung leakage for an add-on electron MLC. AB - Electron IMRT treatments have the potential to reduce the integral dose due to the limited range of the electrons. However, bremsstrahlung produced in the scattering foils could penetrate an added electron MLC (eMLC), thus producing an unmodulated dose contribution that could become unacceptable in electron IMRT treatments. To limit this bremsstrahlung contribution, the photon MLC (xMLC) was used to track the eMLC, but with a margin to avoid penumbra widening through partial screening of the effective electron source. The purpose of this work was to study the effect of the photon-electron MLC tracking on the electron beam penumbra for different treatment head designs. Both isocentric designs and designs where the eMLC is used close to the patient (proximity geometry) have been analysed using Monte Carlo simulations. At 22.5 MeV energy, a tracking margin of 1 cm was enough to avoid penumbra degradation for a helium-filled isocentric geometry, while air-filled geometries (including proximity geometries) require a 2-3 cm margin. Illustrated by an example of a chest wall treatment by electron IMRT, the use of 1 cm tracking margin will reduce the collimator leakage contribution by a factor of 36 as compared to using a static setting of the photon collimator. PMID- 15798317 TI - Imaging dose management using multi-resolution in CT-guided radiation therapy. AB - In image-guided radiation therapy, megavoltage computerized tomography (MVCT) delivers higher dose to the patient for lower image quality than diagnostic kilovoltage CT (kVCT). One way to reduce the mean imaging dose is to reduce the imaging volume, which is often sufficient for registration and dosimetry purposes. The filtered back projection using truncated data causes artefacts that degrade the image quality. Those artefacts can be effectively reduced by wavelet based multi-resolution analysis (WMRA), in which the detail and approximate information are reconstructed separately to bypass the non-locality of filtered back projection. In this study, WMRA was used to reconstruct local images from both very low-dose kVCT scans from a bench-top tomotherapy unit and MVCT scans from helical tomotherapy. Results show that mean imaging dose can be significantly reduced by imaging a small region of interest. In simulation, the root-mean-square error brought by the truncation is smaller than 1-2% and depends on the level of dose reduction. On the other hand, the same mean dose that would have been delivered by a low-quality global CT can be conformed to a smaller volume to improve the visibility of low-contrast organs and fine structures using WMRA. Organs at risk can be avoided during repeated daily CT imaging when irregular-shaped reconstruction areas are used. WMRA does not involve computationally expensive iterations and is suitable for image-guided radiation therapy where imaging speed is essential. Compared with extrapolation methods, errors are further reduced to improve the detection of low contrast and fine structures. PMID- 15798318 TI - Variable transformation of calibration equations for radiation dosimetry. AB - For radiation dosimetry, dosimetric equipment must be calibrated by using known doses. The calibration is done to determine an equation that relates the absorbed dose to a physically measurable quantity. Since the calibration equation is accompanied by unavoidable uncertainties, the doses estimated with such equations suffer from inherent uncertainties. We presented mathematical formulation of the calibration when the calibration relation is either linear or nonlinear. We also derived equations for the uncertainty of the estimated dose as a function of the uncertainties of the parameters in the equations and the measured physical quantity. We showed that a dosimeter with a linear calibration equation with zero dose-offset enables us to perform relative dosimetry without calibration data. Furthermore, a linear equation justifies useful data manipulations such as rescaling the dose and changing the dose-offset for comparing dose distributions. Considering that some dosimeters exhibit linear response with a large dose-offset or often nonlinear response, we proposed variable transformations of the measured physical quantity, namely, linear- and log-transformation methods. The proposed methods were tested with Kodak X-Omat V radiographic film and BANG polymer gel dosimeter. We demonstrated that the variable transformation methods could lead to linear equations with zero dose-offset and could reduce the uncertainty of the estimated dose. PMID- 15798319 TI - Dose verification of single shot gamma knife applications using VIPAR polymer gel and MRI. AB - This work describes an experimental procedure with potential to assess the overall accuracy associated with gamma knife clinical applications, from patient imaging and dosimetry planning to patient positioning and dose delivery using the automated positioning system of a Leksell Gamma Knife model C. The VIPAR polymer gel-MRI dosimetry method is employed due to its inherent three-dimensional feature and linear dose response over the range of gamma knife applications. Different polymer gel vials were irradiated with single shot gamma knife treatment plans using each of the four available collimator helmets to deliver a maximum dose of 30 Gy. Percentage relative dose results are presented not only in the form of one-dimensional profiles but also planar isocontours and isosurfaces in three dimensions. Experimental results are compared with corresponding Gammaplan treatment planning system calculations as well as acceptance test radiochromic film measurements. A good agreement, within the experimental uncertainty, is observed between measured and expected dose distributions. This experimental uncertainty is of the order of one imaging pixel in the MRI gel readout session (<1 mm) and allows for the verification of single shot gamma knife applications in terms of acceptance specifications for precision in beam alignment and accuracy. Averaging net R(2) results in the dose plateau of the 4 mm and 18 mm collimator irradiated gel vials, which were MR scanned in the same session, provides a crude estimate of the 4 mm output factor which agrees within errors with the default value of 0.870. PMID- 15798320 TI - Reassessing the clinical efficacy of two MR quantitative DSC PWI CBF algorithms following cross-calibration with PET images. AB - Clinical cerebral blood flow (CBF) maps generated through dynamic- susceptibility contrast (DSC) magnetic resonance (MR) perfusion imaging are currently cross calibrated with PET studies. The cross-calibration is achieved by rescaling the MR CBF values so that normal white matter CBF corresponds to 22 ml/100 g/min. Examples are provided in this paper to show how this rescaling procedure changes both the clinical interpretation of CBF maps and the manner by which the performance of a given deconvolution algorithm should be assessed. (i) Singular value decomposition-based (SVD) algorithms produce absolute CBF estimates that are inherently under-estimated for all tissue mean transit times (MTT) but, after rescaling, will generate CBF maps that are over-estimated for MTT >4.8 s. (ii) In principle, frequency-domain modelling techniques are expected to be inherently less sensitive to contrast recirculation biases than the time-domain SVD algorithms. However, it is shown that both CBF algorithms become greatly less sensitive to distortions from recirculation after clinical cross-calibration through rescaling has been performed. It is concluded that, when rescaling procedures are employed, it is relatively more important to develop deconvolution algorithms that produce CBF estimates with accuracies that vary little with MTT than to produce algorithms that provide inherently more accurate CBF estimates, but whose relative accuracy varies significantly with MTT. PMID- 15798321 TI - Parallel statistical image reconstruction for cone-beam x-ray CT on a shared memory computation platform. AB - Statistical reconstruction methods offer possibilities of improving image quality as compared to analytical methods, but current reconstruction times prohibit routine clinical applications. To reduce reconstruction times we have parallelized a statistical reconstruction algorithm for cone-beam x-ray CT, the ordered subset convex algorithm (OSC), and evaluated it on a shared memory computer. Two different parallelization strategies were developed: one that employs parallelism by computing the work for all projections within a subset in parallel, and one that divides the total volume into parts and processes the work for each sub-volume in parallel. Both methods are used to reconstruct a three dimensional mathematical phantom on two different grid densities. The reconstructed images are binary identical to the result of the serial (non parallelized) algorithm. The speed-up factor equals approximately 30 when using 32 to 40 processors, and scales almost linearly with the number of cpus for both methods. The huge reduction in computation time allows us to apply statistical reconstruction to clinically relevant studies for the first time. PMID- 15798322 TI - A biomagnetic system for in vivo cancer imaging. AB - An array of highly sensitive biomagnetic sensors of the superconducting quantum interference detector (SQUID) type can identify disease in vivo by detecting and imaging microscopic amounts of nanoparticles. We describe in detail procedures and parameters necessary for implementation of in vivo detection through the use of antibody-labelled magnetic nanoparticles as well as methods of determining magnetic nanoparticle properties. We discuss the weak field magnetic sensor SQUID system, the method of generating the magnetic polarization pulse to align the magnetic moments of the nanoparticles, and the measurement techniques to measure their magnetic remanence fields following this pulsed field. We compare these results to theoretical calculations and predict optimal properties of nanoparticles for in vivo detection. PMID- 15798323 TI - A lesion stabilization method for coronary angiography. AB - A method to make a coronary artery segment of interest appear stationary when viewing a sequence of angiographic images is proposed. The purpose of this method is to facilitate the assessment of lesions caused by coronary artery disease by improving detectability. A description of the stabilization algorithm based on template matching is given. Stabilization was performed on 41 clinical coronary angiograms exhibiting various stenoses and was successful in 39/41 cases. A quantitative analysis of stabilization errors was performed by introducing simulated moving vessels of decreasing contrast into sequences of clinical images. PMID- 15798324 TI - Two-dimensional waveform analysis in MR elastography of skeletal muscles. AB - A method for direct determination of anisotropic elastic coefficients using two dimensional shear wave patterns is introduced. Thereby, the symmetry of the wave patterns is approximated by a squared elliptic equation yielding an explicit relation between waveform and elasticity. The method is used to analyse MR elastography wave images of the biceps acquired by a continuous harmonic excitation at the distal tendon of the muscle. Typically V-shaped wave patterns were observed in this type of tissue, which could be well reproduced by the proposed elliptic approximation of the waveform assuming incompressibility and a transverse isotropic model of elasticity. Without additional experiments, the analysis of straightness, slope and interferences of the wave fronts enabled us to deduce two Young's moduli and one shear modulus, which fully describe the anisotropy of the elasticity of muscles. The results suggest strong anisotropy of the living human biceps causing a shear wave speed parallel to the muscle fibres that is approximately four times faster than the perpendicular shear wave speed. PMID- 15798325 TI - Artefacts in the analysis of temporal response functions measured by photon counting. AB - The least-squares (LS) method in fluorescence decay analyses and in time-domain analyses of the diffuse scattering light for data measured by the time-correlated single photon counting (TCSPC) technique is experimentally evaluated, and the artefact in LS analysis for data with different counting statistics is discussed. In single exponential decay analysis, the error of the decay parameter by the LS method is smaller than 10% of the expected true value when the average number of counts per bin (N/k) is more than 1, and the fitting region covers a period on the order of the decay time. In multi-exponential analysis, the decay parameters are sensitively dependent on the counting statistics. In contrast, the fitting by the maximum likelihood estimation (MLE), assuming Poissonian statistics, greatly reduces such dependence of parameters on the counting statistics. In another application, time-domain diffuse scattering measurements, the LS method is only accurate at N/k > 50 (10% error in the absorption coefficient). In particular, the absorption coefficient is largely dependent on the count. In both examples, the problem of stability in the fitting process by MLE still remains: the convergence of the fitting is critically dependent on the selection of initial guesses of the parameters in contrast to the convergence in the LS method. Thus, a hybrid method using the LS method for the determination of the initial guesses is a practical solution to this problem. PMID- 15798327 TI - Externally triggered gating of nuclear medicine acquisitions: a useful method for partitioning data. AB - Physiological gating in nuclear medicine image acquisition was introduced over 30 years ago to subdivide data from the beating heart into short time frames to minimize motion blurring and permit evaluation of contractile parameters. It has since been widely applied in planar gamma camera imaging, SPECT, positron tomography (PET) and anatomical modalities such as x-ray CT and MRI, mostly for cardiac or respiratory investigations. However, the gating capability of gamma cameras and PET scanners can be employed to produce multiply partitioned, statistically independent projection data that can be used in various ways such as to study the effect of varying total acquired counts or time, or administered radioactivity, on image quality and multiple observations for statistical image analyses. Externally triggered gating essentially provides 'something for nothing' as no data are lost and a 'non-gated' data set is easily synthesized post hoc, and there are few reasons for not acquiring the data in this manner (e.g., slightly longer processing time, extra disk space, etc). We present a number of examples where externally triggered gating and partitioning of image data has been useful. PMID- 15798326 TI - Localization of sentinel nodes in breast cancer: novel method and device to help pen marking of active nodes during gamma camera imaging. AB - Gamma camera imaging with Tc-99m marking is a widely used method to locate sentinel lymph nodes (SNs) in breast cancer patients. Prior to SN biopsy, the anterior and lateral location of the SN is marked on the patient's skin using an ink pen. The pen marks guide the surgeon during an operation. However, in many cases the marking is difficult due to limited space under the detectors of a gamma camera. The aim of this study was to improve the pen marking method. Eleven female patients were imaged 3-4 h after injection of Tc-99m labelled Nanocol. Injection was performed to parenchyma surrounding the breast tumour. To facilitate pen marking, two polycarbonate (PC) plates with 40 x 32 holes (spacing=10 mm) were engineered for anterior and lateral side imaging and then installed on the bed of a dual-head gamma camera. Two drops of Tc-99m were placed into the top corners of both the PC plates, in order to trace the corresponding x y coordinates first from the acquired images and then from the plates. After imaging, the x-y coordinates of the SN(s) were determined from the anterior and lateral side images. Subsequently, the location of each SN was marked with an ink pen on the skin through the small holes in the PC plates. According to the surgeon's evaluation, the distance between the marks and the true location of the SNs was 4.5+/-6.9 mm. Measurements with a custom made phantom revealed that the accuracy of the novel method was significantly (P=0.06) higher as compared with the traditional method (2.7+/-3.0 mm versus 9.2+/-3.0 mm). In addition, we were not able to mark the weakest activity (0.02 MBq) with the traditional method. Taken together, the marking process was considerably easier with the novel method, it had better accuracy and sensitivity than the traditional method and the device is simple enough to be adapted for most gamma cameras. PMID- 15798328 TI - A complete distortion correction for MR images: I. Gradient warp correction. AB - MR images are known to be distorted because of both gradient nonlinearity and imperfections in the B0 field, the latter caused either by an imperfect shim or sample-induced distortions. This paper describes in detail a method for correcting the gradient warp distortion, based on a direct field mapping using a custom-built phantom with three orthogonal grids of fluid-filled rods. The key advance of the current work over previous contributions is the large volume of the mapping phantom and the large distortions (>25 mm) corrected, making the method suitable for use with large field of view, extra-cranial images. Experimental measurements on the Siemens AS25 gradient set, as installed on a Siemens Vision scanner, are compared with a theoretical description of the gradient set, based on the manufacturer's spherical harmonic coefficients. It was found that over a volume of 320x200x340 mm3 distortions can be successfully mapped to within the voxel resolution of the raw imaging data, whilst outside this volume, correction is still good but some systematic errors are present. The phenomenon of through-plane distortion (also known as 'slice warp') is examined in detail, and the perturbation it causes to the measurements is quantified and corrected. At the very edges of the region of support provided by the phantom, through-plane distortion is extreme and only partially corrected by the present method. Solutions to this problem are discussed. Both phantom and patient data demonstrate the efficacy of the gradient warp correction. PMID- 15798329 TI - Integrating a MRI scanner with a 6 MV radiotherapy accelerator: dose increase at tissue-air interfaces in a lateral magnetic field due to returning electrons. AB - In the framework of the development of the integration of a MRI-scanner with a linear accelerator, the influence of a lateral, magnetic field on the dose distribution has to be determined. Dose increase is expected at tissue-air boundaries, due to the electron return effect (ERE): electrons entering air will describe a circular path and return into the phantom causing extra dose deposition. Using IMRT with many beam directions, this exit dose will not constitute a problem. Dose levels behind air cavities will decrease because of the absence of electrons crossing the cavity. The ERE has been demonstrated both by simulation and experiment. Monte Carlo simulations are performed with GEANT4, irradiating a water-air-water phantom in a lateral magnetic field. Also an air tube in water has been simulated, resulting in slightly twisted regions of dose increase and decrease. Experimental demonstration is achieved by film measurement in a perspex-air-perspex phantom in an electromagnet. Although the ERE causes dose increase before air cavities, relatively flat dose profiles can be obtained for the investigated cases using opposite beam configurations. More research will be necessary whether this holds for more realistic geometries with the use of IMRT and whether the ERE can be turned to our advantage when treating small tumour sites at air cavities. PMID- 15798330 TI - Use of an amorphous silicon electronic portal imaging device for multileaf collimator quality control and calibration. AB - Multileaf collimator (MLC) calibration and quality control is a time-consuming procedure typically involving the processing, scanning and analysis of films to measure leaf and collimator positions. Faster and more reliable calibration procedures are required for these tasks, especially with the introduction of intensity modulated radiotherapy which requires more frequent checking and finer positional leaf tolerances than previously. A routine quality control (QC) technique to measure MLC leaf bank gain and offset, as well as minor offsets (individual leaf position relative to a reference leaf), using an amorphous silicon electronic portal imaging device (EPID) has been developed. The technique also tests the calibration of the primary and back-up collimators. A detailed comparison between film and EPID measurements has been performed for six linear accelerators (linacs) equipped with MLC and amorphous silicon EPIDs. Measurements of field size from 4 to 24 cm with the EPID were systematically smaller than film measurements over all field sizes by 0.4 mm for leaves/back-up collimators and by 0.2 mm for conventional collimators. This effect is due to the gain calibration correction applied by the EPID, resulting in a 'flattening' of primary beam profiles. Linac dependent systematic differences of up to 0.5 mm in individual leaf/collimator positions were also found between EPID and film measurements due to the difference between the mechanical and radiation axes of rotation. When corrections for these systematic differences were applied, the residual random differences between EPID and film were 0.23 mm and 0.26 mm (1 standard deviation) for field size and individual leaf/back-up collimator position, respectively. Measured gains (over a distance of 220 mm) always agreed within 0.4 mm with a standard deviation of 0.17 mm. Minor offset measurements gave a mean agreement between EPID and film of 0.01+/-0.10 mm (1 standard deviation) after correction for the tilt of the EPID and small rotational misalignments between leaf banks and the back-up collimators used as a reference straight edge. Reproducibility of EPID measurements was found to be very high, with a standard deviation of <0.05 mm for field size and <0.1 mm for individual leaf/collimator positions for a 10x10 cm2 field. A standard set of QC images (three field sizes defined both by leaves only and collimators only) can be acquired in less than 20 min and analysed in 5 min. PMID- 15798331 TI - A priori identifiability of a one-compartment model with two input functions for liver blood flow measurements. AB - An extended dual-input Kety-Schmidt model can be applied to positron emission tomography data for the quantification of local arterial (f(a)) and local portal venous blood flow (f(p)) in the liver by freely diffusible tracers (e.g., [15O]H2O). We investigated the a priori identifiability of the three-parameter model (f(a), f(p) and distribution volume (Vd)) under ideal (noise-free) conditions. The results indicate that the full identifiability of the model depends on the form of the portal-venous input function (c(p)(t)), which is assumed to be a sum of m exponentials convolved with the arterial input function (c(a)(t)). When m>or=2, all three-model parameters are uniquely identifiable. For m=1 identifiability of f(p) fails if c(p)(t) coincides with tissue concentration (q(t)/Vd), which occurs if c(p)(t) is generated from an intestinal compartment with transit time Vd/f(a). Any portal input, f(p) c(p)(t), is balanced by the portal contribution, f(p) q(t)/Vd, to the liver efflux, leaving q(t) unchanged by f(p) and only f(a) and Vd are a priori uniquely identifiable. An extension to this condition of unidentifiability is obtained if we leave the assumption of a generating intestinal compartment system and allow for an arbitrary proportionality constant between c(p)(t) and q(t). In this case, only f(a) remains a priori uniquely identifiable. These findings provide important insights into the behaviour and identifiability of the model applied to the unique liver environment. PMID- 15798332 TI - Three-dimensional segmentation and skeletonization to build an airway tree data structure for small animals. AB - Quantitative analysis of intrathoracic airway tree geometry is important for objective evaluation of bronchial tree structure and function. Currently, there is more human data than small animal data on airway morphometry. In this study, we implemented a semi-automatic approach to quantitatively describe airway tree geometry by using high-resolution computed tomography (CT) images to build a tree data structure for small animals such as rats and mice. Silicon lung casts of the excised lungs from a canine and a mouse were used for micro-CT imaging of the airway trees. The programming language IDL was used to implement a 3D region growing threshold algorithm for segmenting out the airway lung volume from the CT data. Subsequently, a fully-parallel 3D thinning algorithm was implemented in order to complete the skeletonization of the segmented airways. A tree data structure was then created and saved by parsing through the skeletonized volume using the Python programming language. Pertinent information such as the length of all airway segments was stored in the data structure. This approach was shown to be accurate and efficient for up to six generations for the canine lung cast and ten generations for the mouse lung cast. PMID- 15798333 TI - Localization of the site of origin of reentrant arrhythmia from body surface potential maps: a model study. AB - We have developed a model-based imaging approach to estimate the site of origin of reentrant arrhythmia from body surface potential maps (BSPMs), with the aid of a cardiac arrhythmia model. The reentry was successfully simulated and maintained in the cardiac model, and the simulated ECG waveforms over the body surface corresponding to a maintained reentry have evident characteristics of ventricular tachycardia. The performance of the inverse imaging approach was evaluated by computer simulations. The present simulation results show that an averaged localization error of about 1.5 mm, when 5% Gaussian white noise was added to the BSPMs, was detected. The effects of the heart-torso geometry uncertainty on the localization were also initially assessed and the simulation results suggest that no significant influence was observed when 10% torso geometry uncertainty or 10 mm heart position shifting was considered. The present simulation study suggests the feasibility of localizing the site of origin of reentrant arrhythmia from non invasive BSPMs, with the aid of a cardiac arrhythmia model. PMID- 15798334 TI - Probabilistic two-stage model of cell inactivation by ionizing particles. AB - A model of biological effects of ionizing particles, especially of protons and other ions, is proposed. The model is based on distinguishing the single-particle and collective effects of the underlying radiobiological mechanism. The probabilities of individual particles causing severe damage to DNA, their synergetic or saturation combinations, and the effect of the cellular repair system are taken into account. The model enables one to describe linear, parabolic and more complex curves, including those exhibiting low-dose hypersensitivity phenomena, in a systematic way. Global shape as well as detailed structure of survival curves might be represented, which is crucial if different fractionation schemes in radiotherapy should be assessed precisely. Experimental cell-survival data for inactivation of V79 cells by low-energy protons have been analysed and corresponding detailed characteristics of the inactivation mechanism have been derived for this case. PMID- 15798335 TI - Off-axis chamber response in the depth of photon dose maximum. AB - Measurements as well as Monte Carlo simulations are presented to investigate the deviation between the dose to water and the value measured by an ionization chamber. These deviations are evaluated at different depths (1.5 and 10 cm) and at an off-axis position of 15 cm. It is shown that an ionization chamber can produce a measuring signal, which is up to 2.5% too low, compared to the dose, when measurements are performed at shallow depths and far off-axis. The reason for this underresponse is found in the variation of the wall correction factor. As a result of the variation of the radiation spectra with depth and position the dose to the air volume, which originates from the wall, varies and therefore changes the wall correction factor. PMID- 15798336 TI - Dosimetry of soft x-rays in thin liquid layers. AB - Very thin material layers (<100 microm) partially absorb ionizing radiation of low energy. When irradiating monolayer cell cultures from above, attention must be paid to absorption by the medium. Frequently, the volume of the nutrient medium is variable, and this leads to differences in the radiation doses delivered to the cells. In the present work these conditions were investigated for x-rays of energies between 13 kV and 100 kV in comparison with 60Co gamma rays using chemical dosimetry to measure the absorption by liquid layers between 25 microm and 500 microm thick. When the dose as measured with the ionization chamber was held constant, the dose absorbed in the Fricke solution was shown to increase with decreasing thickness of the layer of liquid because of a dose gradient. The effect of the dose gradient disappeared, however, in thick liquid layers of the Fricke solution by mixing during spectrophotometry. Secondary (photoeffect and Compton) electrons produced in air or filters are responsible for this effect in plastic petri dishes where back scattering at the interface does not occur. This interpretation is suggested by the same results of an analogous experimental setup using gamma rays with a 5-mm-thick Perspex plate. This dose increase in very thin layers, however, could not be verified by irradiating monolayer cells in poured-out plastic petri dishes because the secondary electrons are already absorbed in the remaining liquid film above the cells. PMID- 15798337 TI - Quantitation of the a priori dosimetric capabilities of spatial points in inverse planning and its significant implication in defining IMRT solution space. AB - In inverse planning, the likelihood for the points in a target or sensitive structure to meet their dosimetric goals is generally heterogeneous and represents the a priori knowledge of the system once the patient and beam configuration are chosen. Because of this intrinsic heterogeneity, in some extreme cases, a region in a target may never meet the prescribed dose without seriously deteriorating the doses in other areas. Conversely, the prescription in a region may be easily met without violating the tolerance of any sensitive structure. In this work, we introduce the concept of dosimetric capability to quantify the a priori information and develop a strategy to integrate the data into the inverse planning process. An iterative algorithm is implemented to numerically compute the capability distribution on a case specific basis. A method of incorporating the capability data into inverse planning is developed by heuristically modulating the importance of the individual voxels according to the a priori capability distribution. The formalism is applied to a few specific examples to illustrate the technical details of the new inverse planning technique. Our study indicates that the dosimetric capability is a useful concept to better understand the complex inverse planning problem and an effective use of the information allows us to construct a clinically more meaningful objective function to improve IMRT dose optimization techniques. PMID- 15798338 TI - Effect of patient variation on standard- and hypo-fractionated radiotherapy of prostate cancer. AB - Recent publications suggested that the alpha/beta ratio in the well-known linear quadratic (LQ) model could be as low as 1.5 Gy for prostate cancer, indicating that prostate cancer control might be very sensitive to changes in the dose fractionation scheme. This also suggests that the standard-fractionation scheme based on large alpha/beta ratios may not be optimal for the radio-therapeutic management of prostate cancer. Hypo-fractionated radiotherapy for prostate cancer has received more attention recently as an alternative treatment strategy, which may lead to reduced treatment time and cost. However, hypo-fractionated radiotherapy may be more sensitive to patient variation in terms of disease control than standard-fractionated radiotherapy. The variation of LQ parameters alpha and beta for a patient population may compromise the outcome of the treatment. This effect can be studied by the introduction of the sigmaalpha and sigmabeta parameters, which are the standard deviations of Gaussian distributions around alpha0 and beta0. The purpose of this study is to examine the effect of patient variation in alpha and beta on tumour control probability for standard- and hypo-fractionated radiotherapy of prostate cancer. The tumour control probability based on the LQ model is calculated using parameters alpha, beta, sigmaalpha and sigmabeta. Our results show that sigmaalpha is an important parameter for radiotherapy fractionation, independent of the alpha/beta ratio. A large sigmaalpha will result in a significant increase in the radiation dose required to achieve the same 95% TCP. Compared with the standard-fractionated scheme, sigmaalpha has a smaller effect on hypo-fractionated treatment at lower alpha/beta ratios. On the other hand, for lower alpha/beta ratios, the beta term also plays a more important role in cell-killing and therefore the patient variation parameter sigmabeta must be considered when designing a new dose fractionation scheme. PMID- 15798339 TI - Dosimetric study of a new polymer encapsulated palladium-103 seed. AB - The use of low-energy photon emitters for brachytherapy applications, as in the treatment of prostate or ocular tumours, has increased significantly over the last few years. Several new seed models utilizing 103Pd and 125I have recently been introduced. Following the TG43U1 recommendations of the AAPM (American Association of Physicists in Medicine) (Rivard et al 2004 Med. Phys. 31 633), dose distributions around these low-energy photon emitters are characterized by the dose rate constant, the radial dose function and the anisotropy function in water. These functions and constants can be measured for each new seed in a solid phantom (i.e. solid water such as WT1) using high spatial resolution detectors such as very small thermoluminescent detectors. These experimental results in solid water must then be converted into liquid water by using Monte Carlo simulations. This paper presents the dosimetric parameters of a new palladium seed, OptiSeed (produced by International Brachytherapy (IBt), Seneffe, Belgium), made with a biocompatible polymeric shell and with a design that differs from the hollow titanium encapsulated seed, InterSource103, produced by the same company. A polymer encapsulation was chosen by the company IBt in order to reduce the quantity of radioactive material needed for a given dose rate, and to improve the symmetry of the radiation field around the seed. The necessary experimental data were obtained by measurements with LiF thermoluminescent dosimeters (1 mm3) in a solid water phantom (WT1) and then converted to values in liquid water using Monte Carlo calculations (MCNP-4C). Comparison of the results with a previous study by Reniers et al (2002 Appl. Radiat. Isot. 57 805) shows very good agreement for the dose rate constant and for the radial dose function. In addition, the results also indicate an improvement in isotropy compared to a conventional titanium encapsulated seed. The relative dose (anisotropy value relative to 90 degrees ) from the seed at a distance of 3 cm is close to 70% at 0 degrees whereas that for the titanium encapsulated InterSource103seed is close to 40%. This paper also presents some new Monte Carlo calculations relating to shadowing produced by the seeds in an array implanted for a prostate cancer treatment. Recently, Mobit and Badragan (2004 Phys. Med. Biol. 49 3171) reported shadowing resulting in a 10% decrease in dose from titanium encapsulated 125I seed. We used Monte Carlo simulations (MCNP-4C) to evaluate shadowing for the InterSource103 titanium encapsulated seed and the OptiSeed polymer encapsulated seed. For a specific geometry specified, dose decreases of 13% and 7% were found for the InterSource103 titanium encapsulated and the OptiSeed polymer encapsulated seed, respectively. PMID- 15798340 TI - Modelling simple helically delivered dose distributions. AB - In a previous paper, we described quality assurance procedures for Hi-Art helical tomotherapy machines. Here, we develop further some ideas discussed briefly in that paper. Simple helically generated dose distributions are modelled, and relationships between these dose distributions and underlying characteristics of Hi-Art treatment systems are elucidated. In particular, we describe the dependence of dose levels along the central axis of a cylinder aligned coaxially with a Hi-Art machine on fan beam width, couch velocity and helical delivery lengths. The impact on these dose levels of angular variations in gantry speed or output per linear accelerator pulse is also explored. PMID- 15798341 TI - Fast LROC analysis of Bayesian reconstructed emission tomographic images using model observers. AB - Lesion detection and localization is an important task in emission computed tomography. Detection and localization performance with signal location uncertainty may be summarized by a scalar figure of merit, the area under the localization receiver operating characteristic (LROC) curve, A(LROC). We consider model observers to compute A(LROC) for two-dimensional maximum a posteriori (MAP) reconstructions. Model observers may be used to rapidly prototype studies that use human observers. We address the case background-known-exactly (BKE) and signal known except for location. Our A(LROC) calculation makes use of theoretical expressions for the mean and covariance of the reconstruction and, unlike conventional methods that also use model observers, does not require computation of a large number of sample reconstructions. We validate the results of the procedure by comparison to A(LROC) obtained using a gold-standard Monte Carlo method employing a large set of reconstructed noise samples. Under reasonable simulation conditions, our theoretical calculation is about one to two orders of magnitude faster than the conventional Monte Carlo method. PMID- 15798342 TI - Statistical image reconstruction for transmission tomography using relaxed ordered subset algorithms. AB - Statistical reconstruction methods offer possibilities for improving image quality as compared to analytical methods, but current reconstruction times prohibit routine clinical applications in x-ray computed tomography (CT). To reduce reconstruction times, we have applied (under) relaxation to ordered subset algorithms. This enables us to use subsets consisting of only single projection angle, effectively increasing the number of image updates within an entire iteration. A second advantage of applying relaxation is that it can help improve convergence by removing the limit cycle behaviour of ordered subset algorithms, which normally do not converge to an optimal solution but rather a suboptimal limit cycle consisting of as many points as there are subsets. Relaxation suppresses the limit cycle behaviour by decreasing the stepsize for approaching the solution. A simulation study for a 2D mathematical phantom and three different ordered subset algorithms shows that all three algorithms benefit from relaxation: equal noise-to-resolution trade-off can be achieved using fewer iterations than the conventional algorithms, while a lower minimal normalized mean square error (NMSE) clearly indicates a better convergence. Two different schemes for setting the relaxation parameter are studied, and both schemes yield approximately the same minimal NMSE. PMID- 15798343 TI - Helical cardiac cone beam CT reconstruction with large area detectors: a simulation study. AB - Retrospectively gated cardiac volume CT imaging has become feasible with the introduction of heart rate adaptive cardiac CT reconstruction algorithms. The development in detector technology and the rapid introduction of multi-row detectors has demanded reconstruction schemes which account for the cone geometry. With the extended cardiac reconstruction (ECR) framework, the idea of approximate helical cone beam CT has been extended to be used with retrospective gating, enabling heart rate adaptive cardiac cone beam reconstruction. In this contribution, the ECR technique is evaluated for systems with an increased number of detector rows, which leads to larger cone angles. A simulation study has been carried out based on a 4D cardiac phantom consisting of a thorax model and a dynamic heart insert. Images have been reconstructed for different detector set ups. Reconstruction assessment functions have been calculated for the detector set-ups employing different rotation times, relative pitches and heart rates. With the increased volume coverage of large area detector systems, low-pitch scans become feasible without resulting in extensive scan times, inhibiting single breath hold acquisitions. ECR delivers promising image results when being applied to systems with larger cone angles. PMID- 15798344 TI - Fusion of respiration-correlated PET and CT scans: correlated lung tumour motion in anatomical and functional scans. AB - Lower lobe lung tumours in particular can move up to 2 cm in the cranio-caudal direction during the respiration cycle. This breathing motion causes image artefacts in conventional free-breathing computed tomography (CT) and positron emission tomography (PET) scanning, rendering delineation of structures for radiotherapy inaccurate. The purpose of this study was to develop a method for four-dimensional (4D) respiration-correlated (RC) acquisition of both CT and PET scans and to develop a framework to fuse these modalities. The breathing signal was acquired using a thermometer in the breathing airflow of the patient. Using this breathing signal, the acquired CT and PET data were grouped to the corresponding respiratory phases, thereby obtaining 4D CT and PET scans. Tumour motion curves were assessed in both image modalities. From these tumour motion curves, the deviation with respect to the mean tumour position was calculated for each phase. The absolute position of the centre of the tumour, relative to the bony anatomy, in the RCCT and gated PET scans was determined. This 4D acquisition and 4D fusion methodology was performed for five patients with lower lobe tumours. The peak-to-peak amplitude range in this sample group was 1-2 cm. The 3D tumour motion curve differed less than 1 mm between PET and CT for all phases. The mean difference in amplitude was less than 1 mm. The position of the centre of the tumour (relative to the bony anatomy) in the RCCT and gated PET scan was similar (difference <1 mm) when no atelectasis was present. Based on these results, we conclude that the method described in this study allows for accurate quantification of tumour motion in CT and PET scans and yields accurate respiration-correlated 4D anatomical and functional information on the tumour region. PMID- 15798345 TI - Ultrasound phase-contrast transmission imaging of localized thermal variation and the identification of fat/tissue boundaries. AB - We present a new ultrasound technique for registering localized temperature changes in soft tissues. Conversely, small temperature changes may be induced in order to image tissue layers. The concept is motivated by the search for a compact, low cost method for guiding noninvasive thermal therapies; however its utility may extend to a wide range of imaging problems such as tumour imaging in the breast. This method combines ultrasound transmission imaging, planar projection techniques and phase-contrast theory. After outlining the theoretical foundation of the technique, its feasibility is tested by simulating localized heating within homogeneous tissue layers. Success of this imaging method is evaluated as a function of the ultrasound-imaging wavelength for a Gaussian shaped heated region over the frequency range from 0.1 to 2 MHz. Furthermore we simulate two-dimensional image reconstruction from a receiving array. We conclude that thermal phase-contrast imaging in tissues is plausible for detecting the treatment spot in thermal therapies while operating at frequencies below 1 MHz. Additionally, it may also be possible to use the method for noninvasive thermometry. However, thermometry would require operation at higher frequencies at the tradeoff of increased attenuation and higher sensitivity to scattering, which needs to be further explored. PMID- 15798346 TI - The use of detective quantum efficiency (DQE) in evaluating the performance of gamma camera systems. AB - The imaging properties of an imaging system can be described by its detective quantum efficiency (DQE). Using the modulation transfer function calculated from measured line spread functions and the normalized noise power spectrum calculated from uniformity images, DQE was calculated with the number of photons emitted from a plane source as a measure for the incoming SNR2. Measurements were made with 99mTc, using three different pulse height windows at 2 cm and 12 cm depths in water with high resolution and all purpose collimators and with two different crystal thicknesses. The results indicated that at greater depths a 15% window is the best choice. The choice of collimator depends on the details in the organ being investigated. There is a break point at 0.5 cycles cm-1 and 1.2 cycles cm-1 at 12 cm and 2 cm depths, respectively. A difference was found in DQE between the two crystal thicknesses, with a slightly better result for the thick crystal for measurements at 12 cm depth. At 2 cm depth, the thinner crystal was slightly better for frequencies over 0.5 cm-1. The determination of DQE could be a method to optimize the parameters for different nuclear medicine investigations. The DQE could also be used in comparing different gamma camera systems with different collimators to obtain a figure of merit. PMID- 15798347 TI - Non-invasive high-frequency vascular ultrasound elastography. AB - Non-invasive vascular elastography (NIVE) was recently introduced to characterize mechanical properties of superficial arteries. In this paper, the feasibility of NIVE and its applicability in the context of high-frequency ultrasound imaging is investigated. First, experiments were performed in vitro on vessel-mimicking phantoms. Polyvinyl alcohol cryogel was used to create two double-layer vessels with different mechanical properties. In both cases, the stiffness of the inner layer was made softer. Radial stress was applied within the lumen of the phantoms by applying incremental static pressure steps with a column of a flowing mixture of water-glycerol. The vessel phantoms were insonified at 32 MHz with an ultrasound biomicroscope to provide cross-section sequences of radio-frequency (RF) ultrasound data. The Lagrangian speckle model estimator (LSME) was used to assess the two-dimensional-strain tensors, and the composite Von Mises elastograms were computed. A new implementation of the LSME based on the optical flow equations was introduced. Deformation parameters were estimated using an inversion algorithm. For each in vitro experiment, both layers of approximately 1 mm were distinguished. Second, the use of the method for the purpose of studying small vessels (MicroNIVE) in genetically engineered rodents was introduced. Longitudinal scans of the carotid artery were performed at 40 MHz. The in vivo results give confidence in the feasibility of MicroNIVE as a potential tool to non-invasively study the impact of targeted genes on vascular remodelling in rodents. PMID- 15798348 TI - Neurorehabilitation of the multifaceted and complicated neurologic problems associated with lightning and electrical injuries. PMID- 15798349 TI - Spectrum of neurologic complications of lightning injuries. AB - Lightning is responsible for a wide spectrum of neurologic complications. Many of the persistent symptoms require long-term neurorehabilitation. The lightning related neurologic conditions are divided into four categories. Category I consists of signs and symptoms that are temporary and usually benign. Category II conditions are prolonged or permanent produced by significant central nervous system lesions. Two of the most devastating complications are lightning-related encephalopathy and myelopathy. The disabilities that accompany these lesions call for concentrated neurorehabilitation. A large number of patients are afflicted with neurobehavioral symptoms that may last for months or years. Category III contains delayed neurologic syndromes. Category IV encompasses neurologic lesions that are not directly activated by the lightning strike but are the result of trauma secondary to falls or blasts effects. The sensible course for individuals at risk is to learn and follow lightning safety recommendations. PMID- 15798350 TI - Neurorehabilitation of spinal cord injuries following lightning and electrical trauma. AB - While spinal cord injuries caused by lightning strike or electrical shock are rare, their clinical manifestations pose unique challenges to the clinician who must anticipate the interaction of multiple system involvement with the altered physiology of spinal cord injury. Spinal cord damage may be secondary to the direct effects of electrical current passing through neural tissue producing immediate or delayed impairment. Alternatively, lightning strike and electrical shock may lead to spinal cord damage due to the secondary consequences of injury such as spinal fractures sustained after a fall. In addition to effects on the spinal cord, electrical trauma may result in injury to the brain, peripheral nervous system, musculoskeletal system, skin, and cardiovascular system. This article will review the neurorehabilitation approach to this rare and challenging group of patients. PMID- 15798351 TI - Neurorehabilitation of cerebral disorders following lightning and electrical trauma. AB - The most devastating casualties in lightning and electrical trauma patients are the result of lesions of the nervous system, and especially lesions of the brain. The brain injuries can be divided into three categories: global dysfunction; focal brain injuries; and behavioral-cognitive sequelae without gross physical signs. Lightning injuries are usually the result of outdoor sports and leisure activities. Most electrical trauma cases are the result of workplace accidents. Rehabilitation planning should begin early after the incident and often needs to be continued for a long time. The goal of the rehabilitation team is to maximize functional return in patients with deficits related to brain lesions. The neurorehabilitation team includes the neurorehabilitation physician, physical therapists, occupational therapists, psychologists, speech therapists, and case managers. PMID- 15798352 TI - Autonomic nervous system dysfunction in lightning and electrical injuries. AB - Autonomic nervous system (ANS) dysfunction is a serious complication of lightning and electrical trauma (L/ET). The ANS regulates the normal vegetative functioning of many organ systems. When ANS is compromised after lightning and electric trauma, patients are vulnerable to serious medical problems. Three conditions of ANS dysfunction of particular concern for L/ET patients are complex regional pain syndrome (CRPS), cardiovascular abnormalities, and keraunoparalysis (KP). The patient with CRPS presents with pain, hyperpathia, sweating, and edema hours to days after trauma. Neurorehabilitation is exceedingly important. A primary goal is to keep the affected extremity mobile and functional. Some patients benefit from sympathetic blockade. Cardiovascular abnormalities associated with lightning and electrical trauma can be life threatening. Care for these patients require a multidisciplinary team including a cardiologist. Keraunoparalysis is a frightening and distressing complication of lightning strikes. The syndrome consists of limb paralysis, sensory symptoms, pallor, coolness and absent pulses. Release of excessive catecholamines is said to be responsible for these findings. Fortunately, the condition is transient. PMID- 15798353 TI - Neurorehabilitation of behavioral disorders following lightning and electrical trauma. AB - Neurobehavioral problems after lightning and electrical injuries are diverse. Commonly reported are decreased cognitive function, pain syndromes, depression, posttraumatic stress disorder, and significant alterations in social and work roles. While the problems resemble those following other kinds of accidents, the injury scenarios for lightning and electrical trauma are unique, and seem to invite more skepticism and controversy in medical and legal realms when the survivors seek help. Studies of lightning and electrical injuries have identified disabling neuropsychiatric changes for some survivors, often persistent and occasionally progressive, that appear weakly related to litigation status, inconsistently related to injury scenarios, and likely influenced by individual premorbid emotional and coping patterns. Standards of care in the fields of brain injury, behavioral medicine, and psychotherapy can inform rehabilitation strategies. Proper assessment is important, as well as an individualized approach to treatment. Multidisciplinary intervention focuses on managing symptoms, learning compensatory skills, providing psychosocial support, and preventing maladaptive behaviors. It has been therapeutic for some patients to become activists for better awareness and prevention. PMID- 15798355 TI - Lightning Strike and Electric Shock Survivors, International. AB - Prior to1989 no known support group existed for victims of lightning or electrical shock injury and their families. A lightning strike survivor of twenty years, Steve Marshburn, recognized the need both from his personal experience and after meeting two other survivors. With his wife and psychologist, he formed Lightning Strike and Electric Shock Survivors International. The organization has grown nearly one hundredfold from the thirteen who attended the first annual meeting. The support group serves people from around the world, has produced a number of publications, participated in research about these injuries and has become active in prevention activities. This paper will describe some of the factors that have led to such success for the organization. PMID- 15798354 TI - Rehabilitation of burn injured patients following lightning and electrical trauma. AB - Electrical burn injuries are complicated because of damage to many structures including: blood vessels, muscles, nerves, tendons, bone and skin. Surgeons must confront many problems such as wound healing coverage, scarring, loss of nerve and tendons, progressive joint stiffness and amputation. The goals of burn therapists are to achieve wound healing, functional recovery, and good cosmetic results. Rehabilitation is both preventive and therapeutic and is a fundamental part of managing these patients. In this article, rehabilitation is discussed with emphasis on the following: pain management, wound coverage, positioning, splinting, and exercises (range-of-motion and ambulation). The treatment and prevention of hypertrophic scarring is evaluated. Finally, the use of engineering and assistive technologies for rehabilitation of the electrical burn injured patient is discussed. Successful management of electrical burn injured patients involves communication among the different burn specialists, such as surgeons, anesthesiologists, neurologists, nurses, and kinesitherapists; engineers from the garments/prostheses companies; psychologists; and the patient him/herself. PMID- 15798356 TI - Roadblocks to return to work after electrical trauma. PMID- 15798358 TI - Reduction of spastic tone increase induced by peripheral repetitive magnetic stimulation is frequency-independent. AB - Repetitive magnetic stimulation is able to reduce spastic tone increase after applying it at the spinal cord. It was also found to induce frequency-dependent changes in H-reflex. In a small pilot investigation we stimulated a patient with severe spasticity of the lower limbs and applied different stimulation frequencies. The aim was to look for a clinical measurable effect of spastic reduction depending on the applied frequency. We were unable to show such a dependency, but could confirm the spastic-reducing effects of a peripheral repetitive magnetic stimulation using supratheshold intensities. PMID- 15798357 TI - Biophysical injury mechanisms associated with lightning injury. PMID- 15798359 TI - Letter to the editors. PMID- 15798360 TI - Vision restoration therapy (VRT) efficacy as assessed by comparative perimetric analysis and subjective questionnaires. AB - PURPOSE: We wished to evaluate the efficacy of vision restoration therapy (VRT) in patients with post-chiasmatic brain damage using different functional perimetric tests. These were compared with measures of subjective vision and reaction time. METHODS: An open trial was conducted with hemianopia/scotoma (n=16) patients. Before and after 6 months of VRT results of high resolution (HRP) and Tuebingen automated perimetry (TAP) were evaluated and compared to performance in a Scanning Laser Ophthalmoscope (SLO) as previously reported. Whereas TAP and HRP used above-threshold or near-threshold individual target stimuli on grey background, the SLO used a psychophysical task of detection of three black targets (reverse stimulus) on bright red, patterned background. Subjective testimonials of activities of daily living (ADL) were probed with questionnaires and interviews. RESULTS: Before VRT, the visual field border as assessed by SLO was located significantly closer to the vertical midline than the HRP and TAP border (border mismatch). After VRT the SLO border was still unchanged whereas HRP measurements revealed significant border shifts due to improved stimulus detection (p<0.0001) and improved reaction time (p<0.005) . Fewer misses were also observed in both eyes with TAP (p<0.01) which was primarily due to a significant shift of the absolute borders. Thus, VRT potentiated the mismatch between the SLO borders and the HRP/TAP borders. Fixation performance and the blind spot position remained unchanged after VRT. ADL ratings in the questionnaire improved significantly after VRT which was confirmed by independent patient testimonials. CONCLUSIONS: We replicated earlier findings that VRT improves stimulus detection in HRP and TAP perimetry which were accompanied by subjective, visual improvements. These changes are not caused by fixation or eye movement artifacts. Because the SLO border was located significantly closer to the vertical midline before VRT ("border mismatch") and, in contrast to HRP and TAP, did not change after VRT, we interpret this border mismatch to indicate that the SLO task was too difficult to perform and thus insensitive to VRT effects. Significant reaction time improvements indicate that plasticity of temporal processing might play an important role in vision restoration after brain damage. A further description of the precise psychophysical nature of the restored areas of residual vision is now warranted. PMID- 15798361 TI - BG01V: a variant human embryonic stem cell line which exhibits rapid growth after passaging and reliable dopaminergic differentiation. AB - PURPOSE: To explore a karyotypically abnormal variant human embryonic stem cell (hESC) line, BG01V, as a potential model for studies of dopaminergic neuronal differentiation. METHODS: The properties of BG01V cells were compared to those of normal BG01 cells using immunocytochemistry, RT-PCR, focused microarrays and in vitro differentiation, including dopaminergic differentiation, by culturing with the stromal cell line PA6. RESULTS: Despite the karyotypic abnormality (49, +12, +17 and XXY), undifferentiated BG01V cells expressed pluripotent ESC markers similar to BG01 cells, and retained the ability to differentiate into cell types characteristic of all three germ layers. When co-cultured with the stromal cell line PA6, BG01V cells differentiated into dopaminergic cells which exhibited properties similar to those of mature dopaminergic neurons. CONCLUSIONS: BG01V cells were easier to maintain in culture than karyotypically normal BG01 cells and can be used as an alternative pluripotent hESC type for in vitro developmental studies. PMID- 15798363 TI - Word learning can be achieved without feedback: implications for aphasia therapy. AB - PURPOSE: Children acquire new words through exposure, without the necessity for explicit feedback by caregivers. In aphasia therapy, feedback to the patient is considered an important asset even though the empirical base demonstrating superior learning with online feedback is lacking. The present study examined if healthy adults and patients with chronic aphasia can acquire a new lexicon by intense frequency of exposure alone. METHODS: We compared learning rates with "frequency of exposure alone" (no-feedback condition: n=19 healthy adults; two patients with chronic Broca's and Wernicke's aphasia, respectively) with a condition where subjects additionally received online feedback (feedback condition; n=19). The learning principle was higher statistical co-occurrences of "correct" picture-pseudoword pairings as compared to "incorrect" pairings. In the feedback condition, immediate online feedback on the correctness of respective choices was additionally provided. RESULTS: Both healthy groups successfully acquired the vocabulary. Feedback led to a slight initial acceleration of learning but did not improve latency to peak performance or long-term retention of lexical knowledge. These findings show that high frequency interactive exposure is a potent word learning mechanism in adults and that feedback is not crucial. This is further corroborated by our successful training of two patients with chronic aphasia without online feedback. CONCLUSIONS: Our findings demonstrate that word re-learning in aphasia could benefit from maximizing on the frequency of exposure and exploiting the therapeutic principle of "massed practice", which has been successful in physical rehabilitation after stroke. Additionally, economizing on feedback may prevent patients becoming discouraged by continuous confrontation with their deficits. PMID- 15798362 TI - In vivo electrical stimulation of rabbit retina with a microfabricated array: strategies to maximize responses for prospective assessment of stimulus efficacy and biocompatibility. AB - PURPOSE: Our primary goal was to assess the effects of varying stimulus parameters on the electrically evoked cortical potentials (EECPs) in rabbits, which we intend to use as one measure of biocompatibility of implanted retinal prosthetic devices. We also sought to exclude contamination of waveforms recorded over the occipital cortex from electrical activity from the retina and the degree of reproducibility of EECP recordings. METHODS: A concentric bipolar platinum electrode or microfabricated 5x5 electrode array delivered current to the retina of 43 Dutch-belted rabbits while the EECP was recorded from extradural electrodes over the occipital cortex. Electroretinogram (ERG) and visual evoked cortical potential (VECP) recordings were routinely obtained. Verification that occipital cortical recordings were not heavily contaminated by electrical potentials from the retina (i.e. the "validity" of the cortical recordings) was made by recording retinal and brain responses before and after intravitreal injection of tetrodotoxin. Electrical stimulation of the retina was performed with monopolar (with distant return) or bipolar electrode configurations. Cortical responses were computer-averaged over 100-500 stimulations. The effect of variation in stimulus current, charge, duration, frequency, polarity and spatial orientation of stimulating electrodes on cortical responses was studied. RESULTS: Progressive reduction of responses toward the anterior skull and abolition of posterior recordings by tetrodotoxin indicated that retinal activity did not significantly contaminate EECP recordings. Reproducibility testing revealed that inter-animal variability within the first hour of testing across all animals was not significantly greater than that found during prolonged testing of a single animal. The lowest current that yielded a reproducible EECP with monopolar stimulation was 75 microA (total current through 21 electrodes) using 200 microsec pulses, which yielded a 45 microV cortical response. Strength-duration curves were generally flat for fixed charge stimulation and linear for fixed current stimulation, at least up to a saturation point, which occurred at very high charge. Over 0.5-16 Hz stimulus frequencies, ERGs varied little but evoked potential responses showed a monotonic decline in amplitude at higher frequencies. Large negative-going initial pulses of a biphasic pair yielded the largest cortical amplitudes. EECP amplitudes varied significantly with the orientation of stimulating electrodes on the retina. CONCLUSIONS: This study provides novel data on the reproducibility of EECP recordings, and insight into stimulation parameters that affect retinal and cortical responses. This information can be used to improve the yield of retinal and evoked potential recordings, which will enhance the prospective assessment of the efficacy of stimulation and health of the stimulated tissues following. PMID- 15798365 TI - Improving recovery of spinal cord-injured rats by telomerase-driven human neural progenitor cells. AB - PURPOSE: Human neural progenitor cells hold great promise for treating a variety of human neurological diseases such as spinal cord injury. One of the issues limiting this technology is how to expand neural progenitor cells in vitro to obtain sufficient number of cells for clinical transplantation. We have established a homogeneous population of human neural progenitor cells (hNPC-TERT) immortalized by the human telomerase reverse transcriptase (hTERT) gene. Then we studied whether these cells could differentiate into neural cells in vivo and improve the recovery of spinal cord-injured rats. METHODS: The hNPC-TERT cells had been transplanted into the injured spinal cord and the functional recovery of the rats with spinal cord contusion injury was evaluated through BBB locomotor scale and Motor Evoked Potentials. Additionally, the differentiation of hNPC-TERT cells was shown by immunocytochemistry. RESULTS: As revealed by this animal model, hNPC-TERT cells developed into functional cells in the injured spinal cord and improved recovery from spinal cord injury in both locomotor scores and electrophysiological parameter in this animal model. CONCLUSIONS: This study is the first demonstration of the use of telomerase-driven human progenitor cells to treat spinal cord injury and should provide a new cell source for research of clinical application. PMID- 15798364 TI - Bromodeoxyuridine increases multipotency of human bone marrow-derived stem cells. AB - PURPOSE: Recent reports show that marrow derived mesenchymal stem cells (MeSCs) may have the ability to differentiate into diverse cell types unrelated to their phenotypical embryonic origin, including neural cells. While demonstrated "in vitro" and neonatally, efforts to demonstrate this ability in adult animal brains have had limited success. If it can be shown that human MeSC (HMeSC) can differentiate into neural cells in adult brain, it would open up the possibility that HMeSCs may be of potential therapeutic use in cell replacement therapies for neurological diseases. Here, we demonstrate that adult HMeSCs treated with 5 bromo-2-deoxyuridine (BrdU) for 3 weeks develop the capability to differentiate into neural and retinal cells when provided the appropriate lineage specific differentiation signals in vitro and in adult animals. HMeSC without BrdU treatment did not differentiate into neurons in vitro or adult animal or retinal cells in adult animal. METHODS: MeSCs isolated from adult human bone marrow were treated with BrdU (3 muM) for 3 weeks and then subjected to differentiation conditions both in vitro and in vivo. RESULTS: BrdU pretreated HMeSCs express neuronal and glial markers after co-culture with differentiated human neural stem cells and after transplantation into the adult rat brain. HMeSCs pretreated with BrdU and transforming growth factor-beta3 express a photoreceptor marker after transplantation into the adult rat vitreous. CONCLUSIONS: These results suggest that BrdU treatment may increase the multipotency of HMeSCs for possible use in autologous cell therapies for neurological and opthamological diseases. PMID- 15798366 TI - When and where to publish important findings: a casualty of biogerontology's rise to respectability. PMID- 15798367 TI - Selective mitochondrial autophagy, or mitophagy, as a targeted defense against oxidative stress, mitochondrial dysfunction, and aging. AB - In autophagy, portions of cytoplasm are sequestered into autophagosomes and delivered to lysosomes for degradation. Long assumed to be a random process, increasing evidence suggests that autophagy of mitochondria, peroxisomes, and possibly other organelles is selective. A recent paper (Kissova et al., J. Biol. Chem. 2004;279:39068-39074) shows in yeast that a specific outer membrane protein, Uth1p, is required for efficient mitochondrial autophagy. For this selective autophagy of mitochondria, we propose the term "mitophagy" to emphasize the non-random nature of the process. Mitophagy may play a key role in retarding accumulation of somatic mutations of mtDNA with aging. PMID- 15798368 TI - Mitochondrial DNA gene therapy: a gene therapy for aging? AB - Mutations in mitochondrial DNA cause a group of diverse diseases that affect an estimated half a million people worldwide. These disorders are remarkably resistant to conventional treatments, and thus several gene therapy approaches are being explored. As some of these approaches develop towards maturity, one can't help thinking that some day they may be used against a much more common health problem currently affecting about 6 billion people- aging, which also has been quite resistant to treatment. Unfortunately, we still do not know whether mtDNA mutations significantly contribute to the aging process or not. The prospect of success in mtDNA gene therapy makes getting the answer a high priority. PMID- 15798369 TI - Novel routes for metabolism of 7-ketocholesterol. AB - Oxysterols (oxygenated forms of cholesterol) are present at low levels in the circulation and accumulate is plasma and tissues in some pathologies. In atherosclerotic lesions, 7-oxygenated oxysterols, predominantly 7 ketocholesterol, accumulate and have been implicated in the pathology of the disease. Therefore, knowledge of the mechanisms for 7-ketocholesterol generation and metabolism may provide therapeutic drug targets. There is some in vivo and in vitro evidence that sterol 27-hydroxylase acts on 7-ketocholesterol to initiate its degradation to more polar, water-soluble products. Recent studies indicate an alternative mechanism, in which 7-ketocholesterol is reduced to 7 beta hydroxycholesterol by 11 beta-hydroxysteroid dehydrogenase type 1. PMID- 15798370 TI - Reactive oxygen species production in the mitochondrial matrix: implications for the mechanism of mitochondrial mutation accumulation. AB - The vicious cycle theory postulates that typical mitochondrial DNA (mtDNA) mutations cause their host mitochondria to generate more superoxide and other reactive oxygen species (ROS) than do normal mitochondria, thereby promoting the occurrence of additional mtDNA mutations at an ever-accelerating rate. However, nearly all the loss-of-function mtDNA mutations seen in vivo are large deletions, which (as the original statement of the theory indeed noted, though this has been widely overlooked) should not trigger a vicious cycle because they will prevent the assembly of the potentially superoxide-generating enzyme complexes. Consistent with this is the observation that each cell exhibiting loss of mtDNA encoded function in vivo contains copies of a single, evidently clonally expanded, mutant mtDNA species, whereas the vicious cycle theory predicts a spectrum of mutant forms in each cell. Two recent papers, however, unveil a way in which mtDNA mutations could indeed promote ROS production of their host mitochondria. MtDNA mutations probably shift the intramitochondrial NAD(+)/NADH redox couple towards NADH, and this is now shown in vitro to cause ROS production by alpha-ketoglutarate dehydrogenase, an essential enzyme of the TCA cycle. This does not revive the vicious cycle theory, but it has complex implications for the two most plausible more recent theories, known as "survival of the slowest" and "crippled mitochondria." It may also prove to explain other recent observations in mitochondrially mutant cells in vivo. PMID- 15798372 TI - Thermodynamics and information in aging: why aging is not a mystery and how we will be able to make rational interventions. AB - Currently, the aging research field lacks consensus in its focus and methodology. Foundational principles, such as the evolutionary origins and physiological definition of aging, remain controversial. The aim of this paper is to resolve these issues. By applying the concepts of thermodynamics and information in an evolutionary context, the aging phenotype can be derived from first principles. Life uses information storage to maintain its distance from thermodynamic equilibrium. Since it is impossible to make any process 100% efficient, a selective force (i.e., natural selection) is needed to maintain the information's viability. Natural selection operates upon generations, and for reasons discussed subsequently, the somatic body cannot implement an analogous selective process. The aging phenotype we see can be derived from this model along with a number of insights that will enhance our ability to make intelligent and rational interventions. PMID- 15798371 TI - Stable transformation of CHO Cells and human NARP cybrids confers oligomycin resistance (oli(r)) following transfer of a mitochondrial DNA-encoded oli(r) ATPase6 gene to the nuclear genome: a model system for mtDNA gene therapy. AB - Point and deletion mutations and a general depletion of mammalian mitochondrial DNA (mtDNA) give rise to a wide variety of medical syndromes that are refractory to treatment, possibly including aging itself. While gene therapy directed at correcting such deficits in the mitochondrial genome may offer some therapeutic benefits, there are inherent problems associated with a direct approach. These problems are primarily due to the high mitochondrial genome copy number in each cell and the mitochondrial genome being "protected" inside the double-membrane mitochondrial organelle. In an alternative approach there is evidence that genes normally present in the mitochondrial genome can be incorporated into the nuclear genome. To extend such studies, we modified the Chinese Hamster Ovary (CHO) mtDNA located ATPase6 gene (possessing a mutation which confers oligomycin resistance- oli(r)) by altering the mtDNA code to the universal code (U-code) to permit the correct translation of its mRNA in the cytoplasm. The U-code construct was inserted into the nuclear genome (nucDNA) of a wild type CHO cell. The expressed transgene products enabled the transformed CHO cell lines to grow in up to 1000 ng mL(-1) oligomycin, while untransformed sensitive CHO cells were eliminated in 1 ng mL(-1) oligomycin. This approach, termed allotopic expression, provides a model that may make possible the transfer of all 13 mtDNA mammalian protein encoding genes to the nucDNA, for treatments of mtDNA disorders. The CHO mtATPase6 protein is 85% identical to both the mouse and human mtATPase6 protein; these proteins are highly conserved in the region of the oligomycin resistance mutation. They are also well conserved in the regions of the oligomycin resistance mutation of the mouse, and in the region of a mutation found in Leigh's syndrome (T8993G), also called NARP (neurogenic weakness, ataxia, retinitis pigmentosum). It is likely that the CHO oli(r) mtATPase6 Ucode construct could impart oligomycin-resistance in human and mouse cells, as well as function in place of the mutant ATPase subunit in a NARP cell line. Preliminary experiments on human cybrids homoplasmic for the NARP mutation (kindly supplied by D.C. Wallace), transformed with our construct, display an increased oligomycin resistance that supports these suppositions. PMID- 15798373 TI - Genetically modified hairy roots of Withania somnifera Dunal: a potent source of rejuvenating principles. AB - Transgenic hairy roots were induced from Withania somnifera Dunal, by infecting leaf explants with Agrobacterium rhizogenes. Polymerase chain reaction for rol A gene and Southern blot confirmed the integration of T-DNA in the genome. Cultures were grown in Murashige and Skoog solid as well as in liquid medium. The antioxidant activity was assayed in roots grown in solid media and liquid media. Hairy roots grown in liquid media found to possess highly significant activity in 1,1-diphenyl-2-pecryl-hydrazyl radical, beta-carotene linoleic acid model system. The activity was 57.34%, 75.64%, and 93.41% in case DPPH model and 55.3%, 76.3%, and 90.5% in case of b-CLAMS in 25, 50, and 100 mg L(-1) concentration, respectively. In case of hydroxyl radical trapping and brain lipid peroxidation assay, the activity was more significant in hairy roots grown on solid medium in comparison with commercial formulation prepared using normal roots and standard withanaloids. Root extract grown in solid medium has shown 93.2% hydroxyl radical trapping activity at 100 mg L(-1) concentration, and 500 mg L(-1) has shown 83.6% in case of brain lipid peroxidation assay. High-performance liquid chromatography analysis demonstrated the presence of withanaloids in the hairy root extracts. The results of the study clearly indicate that there is enhancement of secondary metabolites in hairy roots, which is indicated through significant enhancement of the antioxidant activity, since these are the major constituents responsible for the activity. This is the first report on the presence of antioxidant principles in genetically modified roots of W. somnifera. These results of the present study may aid in utilization of the W. somnifera hairy roots for its rejuvenating principles. PMID- 15798374 TI - Gerontomodulatory and youth-preserving effects of zeatin on human skin fibroblasts undergoing aging in vitro. AB - Our studies have shown that zeatin, (6-[4-hydroxy-3-methyl-but-2 enylamino]adenine), a cytokinin plant growth factor, has gerontomodulatory, youth preserving and anti-aging effects on serially passaged human adult skin fibroblasts undergoing aging in vitro. There were no immediate negative or toxic effects in terms of cell attachment, cell proliferation, cell survival, cytoskeletal organization, and cellular growth by treatment with zeatin concentrations between 1 and 200 microM. During long-term treatment, cells could be maintained throughout their replicative lifespan in the presence of 40, 80, and 200 microM zeatin, but the optimal concentration of zeatin's anti-aging and youth preserving effects was found to be 80 microM. Life-long serial passaging of human skin fibroblasts in the presence of zeatin resulted in the prevention of cell enlargement, reduction of intracellular debris, prevention of actin polymerization, and enhancement of cellular ability to decompose hydrogen peroxide and to cope with ethanol and oxidative stresses. Most importantly, anti aging and beneficial effects of zeatin were observed without any induction of additional cell proliferation or an increase in the maximum proliferative capacity, thus ruling out any potentially harmful and carcinogenic effects. PMID- 15798375 TI - Rejuvenation Research 2004: a relaunch with a difference. PMID- 15798377 TI - An interview with Brian M. Delaney. Interview by Vicki Glaser. PMID- 15798378 TI - Validated worldwide supercentenarians for 2004. PMID- 15798380 TI - Early virologic failure in a pilot study evaluating the efficacy of therapy containing once-daily abacavir, lamivudine, and tenofovir DF in treatment-naive HIV-infected patients. AB - Previous investigational data using abacavir (ABC), lamuvidine (3TC), and zidovudine has suggested the possibility of triple nucleoside analogue reverse transcriptase inhibitors (NRTI) therapy as an option in the treatment of HIV infection. We performed a pilot study to assess the potency of once daily ABC+ 3TC+ tenofovir (TDF) in the treatment of HIV-infected naive patients. CD4 and HIV viral load (VL) were followed monthly. Patients were considered to be nonresponder/failing if there was no reduction in VL by >/= 2 log(10) by week 8 and/or a rebound in VL after initial suppression. Resistance testing was then obtained. Nineteen patients naive to antiretroviral therapy (3 women and 16 men) were enrolled, of whom, 2 did not return (withdrew from study at week 2). Median VL and CD4 count at baseline were 147,167 copies per milliliter (5.16 log(10); [range, 7650->750,000]) and 277 cells/mm(3) (range, 59-598). Eight patients had VL > 100000 at baseline. Of 17 patients eligible for follow-up, 5 (27%) were responders (virologic success). Twelve patients (63%) were considered nonresponders and/or with virologic failure. The study was prematurely interrupted because of a high rate of treatment failure. Resistance testing available for 11 nonresponders (58%) showed: 2 patients with wild-type, 5 patients with M184V (reducing susceptibility to 3TC and ABC), 4 patients with M184V+K65R (K65R is responsible for reducing susceptibility to ABC, 3TC and TDF), and none with K65R alone. In conclusion, the combination of ABC, 3TC and TDF cannot be recommended for the initial regimen in HIV treatment-naive patients. PMID- 15798381 TI - Differences in presentation and outcome of invasive pneumococcal disease among patients with and without HIV infection in the pre-HAART era. AB - Invasive pneumococcal disease (PD) occurs frequently among HIV-infected patients, but it is unclear whether its manifestations and outcome are different compared to those observed among patients without HIV-1 infection. Because the immune reconstitution that accompanies antiretroviral therapy may change some of these features and because most cases of HIV- 1 infection occur in resource-poor settings of the world where access to antiretroviral agents is limited, we compared PD among patients with and without HIV-1 infection in a North American population before the introduction of highly active antiretroviral therapy (HAART). The records of all pneumococcal cultures processed at this medical center over a period of 20 months were used to identify patients with invasive PD. Hospital records were reviewed for 103 of these patients (52 with and 51 without HIV-1 infection) and demographic, clinical, laboratory, radiographic, and microbiologic information was abstracted and subsequently analyzed. Despite similarities in presenting signs and symptoms, we found a higher incidence of bacteremia but a more favorable outcome with less frequent requirements for intubation and admission to intensive care units and better survival among individuals with HIV infection. Factors such as less advanced age, the presence of fewer comorbid conditions, or a less florid inflammatory response among HIV infected individuals may account for differences in outcome of invasive PD. PMID- 15798382 TI - Body image in patients with HIV/AIDS: assessment of a new psychometric measure and its medical correlates. AB - HIV infection and its treatment can have significant effects on physical appearance and functioning, which can affect self-perceived body image. We assessed the psychometric properties of a newly developed Body Image Scale (BIS), a subjective measure of body image perception in persons with HIV infection, as well as the scale's relationship to disease progression, symptoms, and demographic factors. HIV-positive men (n = 129) and women (n = 21) attending two outpatient HIV clinics were administered the BIS survey along with a one-page questionnaire. A subset (n = 38) were administered the survey on two occasions to assess test-retest reliability. Nearly half of the sample (46%) had AIDS and 25% had a CD4 count below 200 cells/mm(3) within the prior 3 months. The BIS had unidimensional factor structure, good internal consistency reliability (Chronbach alpha = 0.91), and good test-retest reliability (r = 0.71, p < 0.001) after controlling for the length of interval between assessments. Patients' current perception of their body image was worse then what they perceived it to be prior to HIV infection (p < 0.001), but better than their perception of how others view people with HIV (p < 0.001). The presence of symptomatic disease (p < 0.001) and a diagnosis of AIDS (p = 0.02) were associated with a less favorable body image, although laboratory markers of disease progression (CD4 count and plasma HIV viral load) were not. We conclude that the BIS has good construct validity and is a highly reproducible measure of self-perceptive of body image in HIV-infected patients. Further exploration of its relationship to psychological well being, medication adherence and other aspects of medical care is indicated. PMID- 15798383 TI - Anonymous versus confidential HIV testing: client and provider decision making under uncertainty. AB - Six intensive observational studies of HIV testing sites were undertaken in order to improve understanding of confidential and anonymous testing. Three sites offered only confidential testing (a large health maintenance organization's Urgent Care clinic, the same organization's HIV clinic, and a private medical practitioner's office), one offered only anonymous testing (a free clinic), and two offered a choice of confidential or anonymous testing (a thrift shop alternate testing site and a mobile testing unit). Multiple data collection strategies were used including direct field observation, semistructured interviews with clients and providers, and document and policy analysis. Using an organizational/interactional uncertainty framework, this study found that the choice between anonymous and confidential testing is a central aspect of the HIV testing process, that some clients are unclear about the differences between anonymous and confidential testing, that alternate testing sites' providers play a significant role in encouraging confidential rather than anonymous testing in order to further their organization's resource needs and public health goals, and that testing counselors' may consider that some clients prefer anonymous testing because of fear of stigma, discrimination, or loss of privacy. PMID- 15798385 TI - Development and implementation of a quality improvement program for Ryan White Title I care services using a stakeholder-based model. AB - Both medical and community support services for people living with HIV and/or AIDS have been implemented on a widespread basis since the implementation of the 1990 Ryan White CARE Act. However, many services are provided without adequate evaluation or quality assurance, in spite of federal directions to both evaluate and ensure quality. This report details the development and implementation of a quality improvement project to evaluate Ryan White CARE services using a community stakeholder-based effort. The evaluation was consumer rather than administratively driven, including both consumers and providers to define, measure, and improve services. Project phases included: (1) developing service standards for 14 areas of service provided under Title I of the Ryan White CARE Act; (2) creating and implementing a provider/consumer peer site visit instrument to assess agencies' activities in meeting the service standards; and (3) developing a mechanism to improve quality by linking agencies to technical assistance resources in the metropolitan provider community. By involving providers and consumers in evaluation roles, recommendations by peers could serve as the basis for ongoing quality improvement. PMID- 15798384 TI - Knowledge of structured treatment interruption and adherence to antiretroviral therapy. AB - We conducted a survey of 106 HIV-infected patients on antiretroviral therapy at a community hospital in Lancaster, Pennsylvania, to determine the extent of patient knowledge and attitudes about structured treatment interruption (STI) and whether these were factors in adherence to antiretroviral regimens. Thirty-six percent of patients possessed knowledge of STI as a treatment option and four patients reported that they had stopped taking antiretroviral therapy without specific recommendation from their physician based on information they had heard or read about STI. There was no difference in median adherence based on whether a patient was aware of STI, however, in the group who had heard of STI, attitude that STI is very beneficial was correlated with greater adherence to medication. More than one third of HIV-infected patients on antiretroviral therapy possessed knowledge of STI, and this knowledge affected adherence to antiretroviral regimens. Providers caring for HIV-infected patients should routinely inquire about patient knowledge of STI as another factor in assessing adherence to antiretroviral therapy. PMID- 15798386 TI - Awareness, acceptance, and action: an emerging framework for understanding AIDS stigmatizing attitudes among community leaders in Barbados. AB - AIDS stigma must be acknowledged and reduced to advance HIV prevention and HIV/AIDS care in a variety of settings worldwide, including in the West Indies where national epidemics are thought to be growing rapidly. Regarded by international health organizations as a formidable barrier to service delivery and receipt, AIDS stigma refers to prejudice and discrimination directed toward people living with HIV/AIDS (PLHA), persons perceived as being at risk for HIV infection, and the informal and formal service providers who care for PLHA. However, there is little evidence of successful antistigma interventions in the literature. Furthermore, beyond studies of willingness in various professions to serve clients or patients with HIV/AIDS, the stigmatizing attitudes and behaviors of service delivery personnel, paraprofessionals and volunteers have been inadequately studied. This paper uses data obtained during an AIDS awareness workshop with sports coaches in Barbados to illustrate principles of an antistigma intervention framework being developed for social service and health personnel. The Awareness/Acceptance/Action Model (AAAM) draws on principles of mindfulness, rooted in ancient Asian traditions, and recently adapted to a range of physical and mental health interventions in Western contexts. Mindfulness techniques encourage awareness of one's current state and environment, acceptance of the implications of one's attitudes and behaviors, and the development of intentional responses rather than habitually patterned reactions. In this initial effort, community leaders were guided through a series of self-reflective exercises focusing AAAM principles on their tendencies toward AIDS stigma, and exploring more compassionate and functional alternatives. PMID- 15798387 TI - HIV tests approved. PMID- 15798388 TI - Sea snails help neuropathic pain. PMID- 15798390 TI - An interview with Thomas Levelli. Interview by Vicki Glaser. PMID- 15798391 TI - Improving consistency of cell-based assays by using division-arrested cells. AB - In this article we describe the use of division-arrested cells for cell-based assays designed for high-throughput screening. Cells are the most critical and variable reagent for cell-based high-throughput screening. The robustness of robotic screening depends on the quality and consistency of cell reagents. We demonstrate that for most cell types commonly used for high-throughput screening, cells can be irreversibly division-arrested by mitomycin C treatment at doses that cause no apparent toxicity or obvious change to the cell signaling properties we measured. Our data also suggest that division-arrested cells perform favorably compared to regular growing cells in reporter and calcium flux assays, two platforms most commonly used in robotic screening. Division arrest technology effectively uncouples the process of cell production from robotic screening and brings the convenience of having quality-approved cell reagent on demand for cell-based high-throughput screening. PMID- 15798392 TI - Application of division arrest technology to cell-based HTS: comparison with frozen and fresh cells. AB - Cell-based functional assays are becoming popular in many HTS laboratories because of recent advances in detection and automation technologies. However, the supply of large amounts of live cells with consistent cellular response for day to-day screening operations over several days/weeks is a tremendous challenge. The high cost of cell culture, labor-intensive nature of the work, and inherent variability in cellular responses from time to time tend to be prohibitive for extensive applications of cell-based assays in HTS. We therefore tested division arrested cells that were prepared in a single batch and frozen at -80 degrees C before use in several cell-based assays and in a robotic screening campaign. Chinese hamster ovary cells expressing a Gq-coupled receptor were analyzed for the agonist-induced intracellular Ca2+ response measured on a fluorescent imaging plate reader. In this case, the division-arrested cells showed consistent agonist induced intracellular Ca2+ concentration response as reflected by signal-to-basal ratio and EC50 even 48 h after cell plating. In comparison, the responses from untreated frozen cells and fresh cells declined significantly approximately 30 h after cell plating. In other cell-based assays tested (cyclic AMP assay, reporter gene beta-lactamase assay, and ion-channel assay), the division-arrested cells performed as well as frozen, or fresh cells. We thus conclude that the use of alternate strategies such as frozen cells or division-arrested cells may alleviate the need for several batches of cell plating each day during HTS while maintaining the desired robotic throughput and assay quality. PMID- 15798393 TI - A microtiterplate-based screening assay to assess diverse effects on cytochrome P450 enzyme activities in primary rat hepatocytes by various compounds. AB - During the development of potential drugs it is useful to identify pharmacological and/or toxicological side effects of a compound as early as possible in order to exclude them from further development for reasons of time and cost. Activation or inactivation of members of the cytochrome P450-dependent monooxygenase system (CYP450) might indicate potential undesired effects of a given compound. However, results using CYP450 assay systems are often inconsistent because of different experimental settings. Therefore, it was the goal of the present study to optimize the CYP450 assay in primary rat hepatocytes with respect to the time point of addition of and duration of exposure to alpha naphthoflavone (ANF) and beta-naphthoflavone (BNF) as well as trans-resveratrol (RES), which have well-described stimulatory and inhibitory effects on CYP450 enzymes of the 1A and 2B family, respectively. Hepatocytes were also treated with putative lipoxygenase (LOX)/cyclooxygenase (COX) inhibitors with unknown impact on CYP450 enzyme activity in order to detect potential side effects. Cells were cultured for up to 7 days on 96-well microtiter plates, and enzyme activity was determined by a conventional fluorescence spectroscopy assay. ANF and BNF, given to the cells after 4 days of culture, stimulated CYP1A and 2B activities significantly in a concentration-dependent fashion after long-term exposure for at least 1 day. However, during short-term exposure for 1-6 h, CYP1A activity was inhibited, while CYP2B was increased weakly by ANF but not BNF. RES inhibited CYP1A activity during short- and long-term exposure without affecting CYP2B activity. From the results it was concluded that primary rat hepatocytes should be cultured for at least 3-4 days but no longer prior to the assay. The assay should be performed at two different time points of exposure, i.e., 6 h for short term and 24 h for long-term exposure. The compounds under investigation should be applied at two different concentrations, e.g., at one time and 10 times higher concentrations, which should be oriented to the ED50, provided it is known for the respective substance. Under these assay conditions the LOX/COX inhibitors tested activated CYP1A enzyme activity in long-term but instead inhibited it in short-term experiments. CYP2B activity was stimulated during short- and long-term exposure. These results indicated drug side effects recommending exclusion of the compounds from the drug developmental process. Hence, in order to assess the pharmacological potential of novel compounds it is adequate to perform both short and long-term experiments to concisely describe the effect of a compound on the CYP450 system. PMID- 15798394 TI - An exciplex-based, target-assembled fluorescence system with inherently low background to probe for specific nucleic acid sequences. AB - A novel detection technique, called ExciProbes, has been developed to proof-of principle level for DNA oligonucleotides. The new approach is based on the use of two short oligonucleotides complementary to a target nucleic acid sequence. Each short-probe oligonucleotide bears the separated parts of a new class of fluorescence detector, an exciplex. These isolated parts of the detector have no inherent signal at the detection wavelength. They are designed to detect biotarget by being assembled by the target itself to give a new molecular entity (the exciplex), with a characteristic fluorescence and very large Stokes shift (typically >150 nm). The technique is not related to fluorescence resonance energy transfer, and can potentially resolve to 1 base pair. ExciProbes can detect single or double mutations in a short sequence of DNA, and can be combined with temperature-filtering to provide allelic discrimination of single nucleotide polymorphism analysis. Compared to other fluorophore systems that have large backgrounds (typically >60%), ExciProbes show backgrounds of <1% under comparable conditions, and can be used with DNA, RNA, or synthetic nucleic acids such as locked nucleic acid. PMID- 15798396 TI - A high-throughput assay for modulators of ligand-gated chloride channels. AB - Invertebrate glutamate-gated chloride channels (GluCls) are important targets for anthelmintics and insecticides such as ivermectin. To facilitate screening for novel GluCl modulators, the Caenorhabditis elegans GluCl alpha2beta channel was chosen as a surrogate for parasite channels not yet cloned, and an inducible stable human embryonic kidney cell line was generated. Functional expression of the alpha2 and beta subunits was confirmed by whole-cell voltage clamp assays. Using this cell line, a high-throughput assay was developed that detects membrane potential changes associated with the activation of GluCls. In this assay, membrane depolarization was quantified via changes in fluorescence resonance energy transfer between two membrane-associated dyes. Robust and reproducible signals were detected in response to addition of glutamate or ivermectin. This assay was used for the screening of over 180,000 samples from natural and synthetic sources. PMID- 15798395 TI - Characterization of a hERG screen using the IonWorks HT: comparison to a hERG rubidium efflux screen. AB - The introduction of parallel patch clamp instruments offers the promise of moderate-throughput, high-fidelity voltage clamp for drug screening assays. One such device, the IonWorks HT (Molecular Devices, Sunnyvale, CA), was evaluated and compared to conventional human ethera- go-go-related gene (hERG) patch clamp data and an alternative functional screen based on rubidium flux. Data generated by the IonWorks HT and rubidium assays were compared to determine if either offered superior predictive value compared to conventional patch clamp. Concentration-effect curves for a panel of known hERG blockers were shifted to higher concentrations on the IonWorks HT compared to conventional voltage clamp determinations. The magnitude of the potency shifts was compound-specific and ranged from no shift (e.g., quinidine) to over 200-fold (astemizole). When the extreme value for astemizole was disregarded, the potency shift for 13 other known reference standards was 12-fold or less, with an average shift of fivefold. The same subset of compounds in the rubidium efflux assay exhibited an average potency shift of 12-fold. To provide a simulation of how the IonWorks HT assay might perform in a single concentration screening mode, a panel of test compounds was evaluated. The IonWorks HT screen did not outperform the rubidium efflux screen in predicting conventional voltage clamp measurements. The most likely explanation appears to rest with variable and compound-specific potency shifts in the IonWorks HT assay. The variable potency shifts make it difficult to select a screening concentration that meets the criterion of a high positive predictive value while avoiding false-positives. PMID- 15798397 TI - Development of a non-radioactive, 384-well format assay to detect inhibitors of the mitogen-activated protein kinase kinase 4. AB - Mitogen-activated protein kinase (MAPK) kinases (MKKs, also called MAPK/extracellular signal-regulated kinase [ERK] kinase [MEK]) are constituents of numerous signal transduction pathways involved in growth, differentiation, and stress response. One of its members, MKK4, directly phosphorylates and activates the c-Jun terminal kinases (also called stress-activated protein kinase [SAPK]) in response to stress and pro-inflammatory cytokines. Recent evidence suggest that control of MKK4 activity may provide a novel approach for the treatment of cancer or as anti-inflammatory therapy. To screen for novel low-molecular-weight inhibitors of MKK4, we established a quantitative, non-radioactive in vitro kinase assay. Human MKK4 was expressed as fusion protein with glutathione S transferase (GST) in Escherichia coli. Co-expression of a constitutive active fragment of the MAPK/ERK kinase kinase-1 yielded active GST-MKK4 using GST-SAPK alpha-kinase-negative (KN) mutant as substrate. We determined the kinetic constants for ATP and GST-SAPK alpha-KN. The apparent Km value for GST-SAPKalpha KN was 3.7 microM, while the apparent Km value for ATP was 0.17 microM. Staurosporine inhibited GST-MKK4 with an IC50 of 70 nM. The kinase assay was adapted to a 384-well non-radioactive format. After the kinase reaction the phosphorylated product was captured onto a streptavidin-coated microtiter plate, and phosphorylation was detected with a europium-labeled anti-phosphotyrosine antibody, which allowed time-resolved fluorescence measurement. PMID- 15798399 TI - Cell biology of the neurovascular unit: implications for drug delivery across the blood-brain barrier. AB - The central nervous system (CNS) neurovascular unit is a dynamic structure consisting of vascular endothelial cells, pericytes, and closely juxtaposed astrocytes and neurons. Contact and communication events between cells of the neurovascular unit regulate CNS development, modulate cerebral blood flow, and influence permeability properties of the blood-brain barrier. Dysregulation of proper neurovascular unit function is linked to many common human CNS pathologies, making it a target for a variety of neurotherapeutic interventions. Furthermore, manipulation of the neurovascular unit to enhance the delivery of drugs to the CNS is an active area of interest. In this review I summarize current data concerning the cell and molecular biology of the neurovascular unit. Additionally, I suggest how manipulation of novel protein components of the neurovascular unit may enhance delivery of neurotherapeutic drugs across the blood-brain barrier. PMID- 15798403 TI - New ultrasonic cystofiberscope for staging bladder tumors. AB - We report on a new ultrasonic cystofiberscope for staging bladder tumors. The endoscope makes it possible to view endoscopic and ultrasound images of bladder tumors at any location, either alternately in full screen or simultaneously on the same monitor, with higher resolution. The cystofiberscope is useful for assessing the depth of bladder tumor invasion accurately. PMID- 15798398 TI - Detection of apoptosis and drug resistance of human breast cancer cells to taxane treatments using quartz crystal microbalance biosensor technology. AB - Taxanes are used for the treatment of many human cancers, as first- and second line chemotherapeutics. In the course of treatment many patients develop resistance or hypersensitivity to one form of taxane and require a different taxane to rescue the therapeutic benefit of the drug. There is currently no method to reliably predict tumor responses to taxanes prior to therapy or when resistance or hypersensitivity develops. We adapted the quartz crystal microbalance (QCM) biosensor technique to study responses of human mammary epithelial tumor cells to taxanes. Studies indicate that stable frequency and resistance levels are reached at 24 h. Cells in the QCM can then be treated with taxanes and responses monitored in real time via frequency and resistance changes reflecting alterations of cell mass distribution and viscoelastic properties. Distinct shifts in frequency and resistance accurately predicted apoptosis or resistance to treatment, as determined in parallel convention assays. QCM analysis accurately predicted docetaxel was more effective than paclitaxel and MCF-7 cells were more resistant to taxanes compared to MDA-MB-231 cells. These studies suggest "signature" patterns for taxane responsivity could be compared to those of patient biopsy samples to predict therapy outcome prior to treatment for initial therapy or to rescue therapy efficacy. PMID- 15798404 TI - Evolution of hand-assisted laparoscopic surgery. PMID- 15798406 TI - Laparoscopic resection and ureteroureterostomy for congenital midureteral stricture. AB - A 16-year-old boy presented with flank pain and was found to have right-sided hydronephrosis and hydroureter on ultrasonography and an intravenous urogram. A retrograde pyelogram revealed a tight, short-segment, non-negotiable stricture in the midureter. A CT scan excluded extrinsic compression. In the absence of any other pathology, the stricture was considered to be congenital. The diseased segment of the ureter was resected laparoscopically, and an intracorporeally sutured ureteroureterostomy was fashioned over a double- J stent. The patient made an uneventful recovery and was well at follow-up 18 months later. To the best of our knowledge, this is the first reported case of laparoscopic resection of a congenital midureteral stricture. PMID- 15798405 TI - Incidental detection of purulent fluid in kidney at percutaneous nephrolithotomy for branched renal calculi. AB - BACKGROUND AND PURPOSE: Some patients undergoing percutaneous nephrolithotomy (PCNL) have purulent fluid in the pelvicaliceal system at the time of puncture, although preoperative features are not suggestive of infection. We report the management and outcome of 19 such patients. PATIENTS AND METHODS: Among the 639 patients who underwent PCNL at our center from July 2000 to October 2003, 19 had purulent fluid in the kidney at initial puncture. The preoperative findings, operative details, hospital course, and final outcome in these patients were analyzed. Twelve patients had stone removal at the first sitting (option 1), while seven had a nephrostomy tube placed initially with stone removal deferred for 3 to 7 days (option 2). RESULTS: With option 1, stones were cleared with one to three tracts in 45 to 120 minutes. Seven patients recovered smoothly, three had transient fever, while one had grade I and another had grade II sepsis. The purulent fluid grew E. coli in two cases and Proteus or Serratia in one case each and was sterile in eight. With option 2, stones were cleared using one to three tracts in 60 to 100 minutes. The fluid grew E. coli or Klebsiella in one case each and was sterile in five. Three patients had a smooth recovery, two had transient fever, one had grade I sepsis, and another had grade II sepsis. Risk factors for sepsis, irrespective of the option, were a recent history of febrile urinary-tract infection, borderline elevation of total leukocyte count, thick or foul pus as opposed to mere turbidity, the use of a single tube or tract or delayed creation of second tracts, and operating time >90 minutes. CONCLUSIONS: Despite normal preoperative urine and blood values, one may find purulence on puncture during PCNL, which is not always infected. If any one or more of the above risk factors is present, it is safer to drain the kidney initially, making sure that all blocked calices are drained, inserting multiple tubes if necessary. PMID- 15798407 TI - Laparoscopic pyeloplasty using endoscopic GIA stapler for ureteropelvic junction obstruction. AB - PURPOSE: We applied laparoscopic pyeloplasty in 10 patients with ureteropelvic junction (UPJ) obstruction. To evaluate the efficiency and safety of this procedure using an endoscopic GIA stapler, the clinical outcomes and our procedures are presented. PATIENTS AND METHODS: From August 1996 to March 2003, eight female and two male patients with a mean age of 22.3 years suffering from UPJ obstruction diagnosed by various combinations of ultrasonography, excretory urography, retrograde ureteropyelography, CT, and MRI were treated with laparoscopic dismembered Anderson-Hynes pyeloplasty with resection of a dilated redundant renal pelvis. In six cases, an endoscopic gastrointestinal automatic stapler (Endo-GIA) was used. The procedure was performed via an extraperitoneal approach in two cases and a transperitoneal approach in eight. RESULTS: Laparoscopic pyeloplasty was successful in all patients, including the six treated using an Endo- GIA stapler. The mean operating time was 291 minutes, and the mean anastomotic time was 105 minutes, with a mean estimated blood loss of 44 mL. Postoperative complications occurred in five cases: anastomotic urinary leakage in two and pyelonephritis in three. The mean time to full convalescence in the entire series was 22 days. No urolithiasis occurred in the patients treated with the Endo-GIA stapler during the follow-up period of 2 to 76 (mean 22) months. CONCLUSIONS: Laparoscopic dismembered pyeloplasty including the Endo GIA stapler technique is an efficient and safe procedure that provides excellent results for extrinsic or complicated UPJ stenosis. The risk of stone formation has not yet been determined. PMID- 15798409 TI - Ureteroscopy in the management of pediatric urinary tract calculi. AB - PURPOSE: To report our experience with ureteroscopy in the treatment of pediatric urinary tract calculi and present a review of the literature. PATIENTS AND METHODS: Between 1988 and 2003, 52 ureteroscopic procedures were performed in 25 male and 10 female children aged 11 months to 15 years (mean 5.9 years). Using a semirigid 6.8F 43-cm ureteroscope and routine antibiotic prophylaxis, stones were fragmented with a pulsed-dye laser (N = 14; stone size 6-15 mm with a mean of 9.6 mm), electrohydraulic lithotripsy (EHL) (N = 26; stone size 3-20 mm with a mean of 8.4 mm), or a holmium laser (N = 7; stone size 5-15 mm with a mean of 10 mm); removed by basket extraction (N = 5; stone size 5-8 mm with a mean of 7 mm); or both. Stenting or ureteral dilatation was not performed routinely. RESULTS: With the pulsed-dye laser, there was an overall stone-free rate of 72%. Complications consisted of one ureteral perforation and one stenosis of the intramural portion of a megaureter (14% complication rate). With EHL, the overall stone-free rate was 92%. Complications consisted of one case each of ureteral perforation and incipient urinary retention and five of mild fever (27%). With the holmium laser, the overall stone-free rate was 100%, and there were no complications. Basketing likewise produced a 100% stone-free rate, and there was one complication, a mucosal tear in a patient who also underwent pulsed-dye laser lithotripsy. CONCLUSION: Ureteroscopy is a safe and effective means of treating the majority of pediatric ureteral calculi, although retreatment rates are higher with multiple stones and in younger children. Dilatation of the vesicoureteral junction is usually not necessary with ureteroscopes <8F, nor is ureteral drainage required after uncomplicated ureteroscopy. The holmium laser is the most effective and safest method of fragmentation regardless of stone composition. Ureteroscopy for this indication should be performed only by an experienced endoscopist. PMID- 15798408 TI - Febrile complications following insertion of 100 double-J ureteral stents. AB - BACKGROUND AND PURPOSE: Implanted foreign bodies are associated with a higher risk of infection. Little has been published on infectious complications associated with ureteral double-J stents. The aim of this study was to define risk factors and characterize the febrile complications of the insertion of double-J stents. PATIENTS AND METHODS: One hundred consecutive cases of retrograde stent insertion (55 men and 45 women) were evaluated retrospectively. Eighty-one patients had an obstructing stone, either in the ureter or at the ureteropelvic junction; 10 had an obstructing tumor; and 9 had hydronephrosis and pain without calculi. Risk assessment was calculated for two major categories: urgent (N = 47) and elective (N = 53) insertion. Potentially confounding factors such as age, sex, accompanying medical problems, and types of prophylaxis were assessed. All but six patients received prophylactic antibiotics prior to stent insertion. RESULTS: All thirteen patients who had fever at the initial evaluation underwent urgent stent insertion. Of the 87 afebrile patients (53 elective and 34 urgent insertions), 22 (25%) developed fever (> or = 38 degrees C) after the procedure. Fever developed in 19 (56%) of the 34 urgent-insertion group who were not originally febrile as opposed to 3 (6%) of the elective-insertion group (P < 0.001). Six of these 22 patients (27%) had a positive urine culture after insertion. None had a positive blood culture, including the 13 patients who underwent the procedure while febrile. CONCLUSIONS: Urgent insertion of a double J stent is associated with a high risk of fever but a favorable outcome. PMID- 15798410 TI - Minimally invasive therapy for benign prostatic hyperplasia: practice patterns in Minnesota. AB - BACKGROUND AND PURPOSE: Benign prostatic hyperplasia (BPH) affects more than 50% of men by the age of 60 and 90% by age 85. Many of these men are not candidates for surgical procedures such as transurethral resection of the prostate (TURP), stimulating the development of less-invasive forms of therapy. We studied the utilization of these newer therapies by urologists practicing in Minnesota. MATERIALS AND METHODS: An anonymous questionnaire was sent to 174 members of the Minnesota Urological Society, of which 58 were available for analysis. A case scenario was presented of a patient with BPH refractory to medical therapy. The options were traditional and minimally invasive therapies. The physician was asked to select whether he or she would offer each option and perform the procedure or refer the patient within or outside the practice. Statistical analysis was performed using chi-square and two-sample t-tests on Minitab software. The results were considered significant at P < 0.05. RESULTS: While 59% of the respondents would offer both minimally invasive and traditional alternatives, 10% would offer only minimally invasive therapy, while 29% would offer only traditional therapy (P = 0.01). The most common minimally invasive therapies offered were transurethral microwave thermotherapy and (55%) and transurethral needle ablation (33%). If they offered a form of minimally invasive therapy, the majority of respondents would perform the procedure themselves. Rural urologists were less likely to offer minimally invasive therapy (43%) than metro physicians (81%; P = 0.035). There was no significant difference in the use of minimally invasive therapies by rural and urban urologists (P = 0.409) or urban and metropolitan urologists (P = 0.119). Urologists completing their training between 1960 and 1980 were less likely to offer minimally invasive therapy. There was no significant difference in the likelihood of offering traditional versus minimally invasive alternatives according to the percent of managed care in the practice. CONCLUSIONS: Urologists closer to the completion of their residency training are more likely to include a minimally invasive technique in their treatment plan, while urologists practicing in rural Minnesota are less likely to offer minimally invasive procedures. Further emphasis should be placed on increasing the availability of minimally invasive techniques in rural settings. PMID- 15798412 TI - Effect of urine specific gravity on effectiveness of shockwave lithotripsy. AB - PURPOSE: To determine the effect of urine specific gravity (SG) and radiographic contrast medium on the efficiency of extracorporeal shockwave lithotripsy (SWL) fragmentation using a previously published in vitro model. MATERIALS AND METHODS: Artificial kidney stones (BegoStone [BEGO USA, Smithfield, RI]) of a standard size (10 x 10 mm) and weight (mean 1.90 g; range 1.89-1.91 g) were randomly allotted to one of four test media with differing SG: group 1: synthetic urine, SG 1.003; group 2: synthetic urine, SG 1.040; group 3: 50% radiographic contrast (Cysto-Conray II, Mallinckrodt Inc., Hazelwood, MO), 50% synthetic urine, SG 1.069; and group 4: 100% contrast, SG 1.121. The stones were placed in cups made of 2.2-mm screen mesh and exposed to 500 shockwaves at 20 KV from a spark-gap lithotripter (Econolith 2000; Medispec, Germantown, MD). The stone fragmentation rate (i.e., percent fragmentation) was determined by dividing the dry residual stone weight by the prelithotripsy dry weight (X 100). RESULTS: The mean fragmentation rate for the four groups was 31.7%, 37.6%, 31.8%, and 27.2%, respectively. Statistical significance was not achieved (P = 0.1). CONCLUSION: Stone breakage tends to be more effective when urine SG is about 1.040 than at higher or lower values. Hence, the recommendation for an overnight fast prior to SWL is well founded. Introduction of contrast medium may inhibit stone breakage because it increases the specific gravity. PMID- 15798411 TI - Randomized double-blind comparison of lidocaine gel and plain lubricating gel in relieving pain during flexible cystoscopy. AB - PURPOSE: To compare the pain induced by outpatient flexible cystoscopy in men having local lidocaine jelly application or plain lubricant. PATIENTS AND METHODS: We performed a randomized, double-blind study to compare the anesthetic effect of intraurethral 2% lidocaine gel (N = 45) and plain lubricant (N = 46) in men undergoing outpatient flexible cystoscopy. The age, prostate size, and examination time were similar in the two groups. Before cystoscopy, we filled the urethra with 20 mL of gel, which was held for 15 minutes. A 15.5F flexible cystoscope was then used for examination. A 10-point visual analog pain scale (1 = least to 10 = most painful) and a four-point pain grade (grade 1 = least to grade 4 = most painful) were used to measure the pain perception. The amount of postoperative analgesic used and willingness to adopt the same anesthesia for future cystoscopy were also recorded. RESULTS: The mean pain scores for the lidocaine and plain gel groups were 2.8 +/- 1.1 and 2.5 +/- 1.1, respectively (P = 0.06), while the pain grades for the lidocaine gel and plain gel groups were 1.6 +/- 0.6 and 1.8 +/- 0.7, respectively (P = 0.19). In both groups, many patients (42.2% in the lidocaine gel group and 37% in the plain lubricant group) felt the external sphincter and prostatic urethra were the most uncomfortable areas during cystoscopy. Postcystoscopic analgesics were requested by 12 of 45 patients in the lidocaine group, but only 6 of 46 of the plain-gel group (P = 0.103). When patients were asked if they would desire general anesthesia for better pain control in future examinations, five of the lidocaine group responded positively, while three of the plain-gel group said they would (P = 0.62). The cost of lidocaine gel is about thrice that of the plain gel. CONCLUSION: Using plain lubricant is cheaper and faster than applying lidocaine gel. The pain score, pain grade, postcystoscopic analgesic requirement, and anesthetics requested for the next cystoscopy were similar in the two groups. However, a larger investigation will be needed to achieve more significant statistical power. PMID- 15798413 TI - Impact of a double-pigtail stent on ureteral peristalsis in the porcine model: initial studies using a novel implantable magnetic sensor. AB - BACKGROUND AND PURPOSE: The effect of stents on ureteral peristalsis in vivo is not entirely clear. We sought to develop a minimally invasive method for its study. MATERIALS AND METHODS: In female domestic pigs, electrical potentials from the ureter were measured by bipolar steel-wire electromyography electrodes delivered laparoscopically. Mechanical movement was measured by giant magneto resistive sensors mounted on custom-made aluminum strips. After baseline values were obtained, the animals were randomized to receive silicone or polyurethane stents, and ureteral peristalsis was measured for 8 hours acutely and for 4 hours 1 week later. RESULTS: Implantation of the devices took an average of 30 minutes. A consistent correlation was found between laparoscopically observed peristaltic waves and the peristalsis detected by the two measuring devices. The devices themselves did not affect peristalsis. Stent insertion increased peristaltic activity initially but later reduced or stopped it. There was no difference in the effects of the two types of stents. CONCLUSIONS: The new technique permits close monitoring of ureteral peristalsis in vivo. Smaller stents appear to have less immediate effect than larger ones, but all type of stents tested eventually caused aperistalsis. PMID- 15798414 TI - Genetic and dietary factors in urinary citrate excretion. AB - BACKGROUND AND PURPOSE: Hypocitraturia, an important risk factor for calcium oxalate nephrolithiasis, is the result of numerous factors. We studied citrate excretion by patients with and without stones consuming normal and controlled formula diets. SUBJECTS AND METHODS: Subjects with and without a history of calcium oxalate stones (N = 101 per group) provided two or three 24-hour urine specimens during consumption of self-selected diets. Data also were collected on subsets of subjects consuming formula (Ensure) diets. Citrate was determined using the citrate lyase method of Petrarulo and associates, and values for multiple specimens were averaged. The data were adjusted for creatinine excretion and examined on a per-day basis. RESULTS: The mean citrate excretion of the non stone formers was slightly but not significantly higher than that of the stone formers (442 +/- 217 versus 378 +/- 153 mg/g of creatinine). All statistical analyses revealed highly significant differences between, but not within, individuals, a result compatible with a genetic influence. In the normal population, 5% of subjects had a citrate excretion <200 mg/g of creatinine, whereas this result was seen in 34% of the stone-forming subjects. When the subjects consumed a formula diet, women in both groups had much higher citrate excretion than when on a self-selected diet, but little difference was seen in the men. The patterns of citrate recovery suggest low, intermediate, and high excretors. In the normal population, 15% of subjects excreted <340 mg/g of creatinine, whereas this was true of 43% of the stone-forming subjects. Analysis of six families suggested three excretor phenotypes, with a codominant pattern of inheritance. CONCLUSION: These findings imply a genetic influence on citrate excretion, as has already been demonstrated for calcium excretion. Further studies of genetic influences on calcium oxalate stone formation are warranted. PMID- 15798415 TI - Obstruction alters the effect of prostaglandin E2 on ureteral contractility. AB - BACKGROUND AND PURPOSE: Although our understanding of ureteral physiology during acute obstruction remains limited, we believe that prostanoids (prostaglandins [PGs], thromboxanes, prostacyclins) play a major role in modulation of ureteral contractility and that inhibition of prostanoid synthesis causes substantial reduction in in-vitro and in-vivo ureteral contractility rates. The purpose of this study was to determine the in-vitro effects of PGE2 on chronically obstructed human and acutely obstructed porcine ureters. MATERIALS AND METHODS: Female pigs underwent unilateral laparoscopic ureteral obstruction. Following 1, 2, 6, 24, and 48 hours of obstruction (n = 3 at all points), animals were euthanized, and obstructed, contralateral nonobstructed, and normal (from unobstructed pigs) ureters were harvested. Chronically obstructed human ureter was obtained from subjects who underwent nephrectomy to remove nonfunctioning kidneys. Normal human ureter was obtained from the discarded portion of excess distal ureter in patients undergoing elective donor nephrectomy. Rings 4 to 5 mm long were suspended in aerated Krebs buffer, and their spontaneous contractions and contraction in response to various concentrations of PGE2 were recorded. RESULTS: Prostaglandin E2 increased contractility in chronically obstructed human ureters. In acutely obstructed porcine ureteral segments, low concentrations of PGE2 inhibited ureteral contractility in a dose-dependent fashion, similar to controls. At higher concentrations of PGE2, contractility was increased. This increase was more pronounced with longer intervals of obstruction in a time dependent manner. CONCLUSION: Prostaglandin E2 increased contractility in obstructed ureters while relaxing normal and nonobstructed ureters. The response to PGE2 was accentuated by a longer duration of obstruction. Prostaglandin E2 may be a unique target for pharmacologic modulation in the treatment of symptoms associated with acute urinary obstruction. PMID- 15798416 TI - Three-dimensional CT virtual endoscopy in the detection of simulated tumors in a novel phantom bladder and ureter model. AB - BACKGROUND AND PURPOSE: Cystoscopy and ureteroscopy have limitations in the evaluation for urothelial tumors, and both are invasive. We studied the utility of three-dimensional (3D) CT virtual endoscopy in phantom models. MATERIALS AND METHODS: A phantom pelvis was constructed of Plexiglas, porcine pelvic bones, and processed animal fat and scanned at various table speeds in a four detector-row CT machine for ability to detect "tumors" of Solidwater plastic polymer. Images were reconstructed at slice thicknesses of 2.5 to 5.0 mm and reconstructed in 3D for evaluation by two radiologists with no knowledge of the scanning parameters or tumor location. Similar studies were performed with a ureter model. RESULTS: With 5-mm slices, the sensitivity for bladder tumors ranged from 67% for 2-mm tumors to 100% for 4-mm tumors, with 12 false-positive findings. The overall sensitivity was 86% with 3.75-mm slices with one false positive, and with 2.5-mm slices, the sensitivity was 93%, again with one false positive. For the ureteral tumors, the overall sensitivities and numbers of false positives were 88.9% and eight with 5.0-mm collimation, 88.9% and four with 3.75-mm collimation, and 100% and three with 2.5-mm collimation. The effective radiation dose for all studies was equivalent to that of a standard abdomen/pelvis scan. CONCLUSIONS: Although virtual endoscopy traditionally has had difficulty detecting tumors <5 mm, the multidetector-row CT protocols used in this study could detect most lesions smaller than this. The scan also depicts the other tissues of the pelvis, which is valuable for staging. The 3D images were produced using data from the CT urogram parameters standard at our institution. PMID- 15798418 TI - Assessment of novel ureteral occlusion device and comparison with stone cone in prevention of stone fragment migration during lithotripsy. AB - PURPOSE: To present the initial ex-vivo results of a new ureteral-occlusion device (Cook Urological, Spencer, IN) for preventing the retrograde migration of particles during ureteroscopy and its comparison with the Stone Cone (Microvasive Boston Scientific Corp., Spencer, IN). MATERIALS AND METHODS: We designed an ex vivo model using a porcine kidney with an intact ureter. Standardized plastic beads were placed in the proximal ureter. Each device was deployed proximal to the beads during ureteroscopy, and the number of beads impeded by each device was recorded. RESULTS: The new device was successful at preventing retrograde migration of beads > or = 1.5 mm. Furthermore, 1.0-mm beads were impeded by the center of the device, although they passed through the periphery. The Stone Cone blocked the retrograde migration of beads > or = 2.5 mm. CONCLUSION: The new occlusion device was efficacious in an ex-vivo model. It appears to prevent the migration of smaller particles than are blocked by the Stone Cone. PMID- 15798420 TI - Dismembered percutaneous endopyeloplasty: a new procedure. AB - PURPOSE: We recently described a novel technique of percutaneous non-dismembered endopyeloplasty (Fenger type). Herein, we extend this transrenal technique further and report percutaneous dismembered endopyeloplasty (Anderson-Hynes type). MATERIALS AND METHODS: In five pigs with unilateral ureteropelvic junction (UPJ) obstruction created 3 to 6 weeks earlier, percutaneous dismembered endopyeloplasty was performed. Percutaneous transrenal access to the UPJ was obtained, and the UPJ was completely dismembered from within the renal pelvis through the solitary percutaneous tract. The dismembered proximal ureter was circumferentially mobilized, and in two animals, the UPJ segment was completely excised and removed. A spatulated end-to-end endopyeloplasty anastomosis (Anderson-Hynes) was created transrenally with 5 to 10 interrupted sutures using a novel nephroscopic suturing device (Sew-Right SR-5; LSI Solutions, Rochester, NY). In two animals, the entire percutaneous procedure was performed with CO2 insufflation instead of fluid irrigation. RESULTS: The technique was developed in three pigs. Subsequently, two pigs were treated and sacrificed at 2 and 5 weeks. All UPJs were dismembered successfully, and a precisely sutured mucosa-to-mucosa anastomosis was created. Intraoperative bleeding was negligible, and the operative time ranged from 3 to 5 hours, with the majority of the time dedicated to transrenal retroperitoneal dissection of the scarred, fibrotic UPJ. Carbon dioxide insufflation was efficacious because it minimized fluid extravasation and tissue edema and additionally enhanced visibility. Postoperative pyelograms revealed an adequately funneled UPJ, with good flow into the distal ureter. The two survival animals had minimal apparent morbidity from the procedure, and retrograde pyelograms at euthanasia revealed a patent anastomosis without extravasation. A 6F catheter easily crossed the reconstructed UPJ at autopsy in all animals. CONCLUSIONS: Dismembered percutaneous Anderson-Hynes endopyeloplasty is technically feasible and is promising. Further technical experience and additional functional outcome analysis in the survival model are necessary. With the technique described herein, we introduce the concept of percutaneous intrarenal reconstructive surgery (PIRS), wherein advanced intrarenal and retroperitoneal dissection with reconstruction can be performed endourologically, further broadening the horizons of conventional percutaneous techniques. PMID- 15798417 TI - Correlation of preoperative three-dimensional magnetic resonance angiography with intraoperative findings in laparoscopic renal surgery. AB - BACKGROUND AND PURPOSE: Contrast-enhanced three-dimensional magnetic resonance angiography (3D-MRA) with image reconstruction has important applications in laparoscopic urologic surgery. We now use 3D-MRA as part of our preoperative evaluation in selected patients undergoing laparoscopic donor nephrectomy, pyeloplasty, radical nephrectomy, and partial nephrectomy. PATIENTS AND METHODS: From June 2001 to December 2002, 50 patients underwent preoperative 3D-MRA at 1.5 T prior to laparoscopic renal surgery. In general, preoperative 3D-MRA was obtained for donor nephrectomies and pyeloplasties and for cases where prior imaging suggested a possible vascular anomaly. Patients who underwent preoperative imaging included those having donor nephrectomy (N = 28), pyeloplasty (N = 12), radical nephrectomy (N = 5), partial nephrectomy (N = 3), and other laparoscopic renal procedures (N = 2). The 3D-MRA studies were interpreted by one radiologist, and all laparoscopic cases were performed by one of two surgeons. The findings of 3D-MRA were correlated with the intraoperative findings with special attention to aberrant vasculature, including duplicated renal arteries or veins, accessory vessels, or crossing vessels. RESULTS: Among patients undergoing laparoscopic donor nephrectomy, 3D-MRA correctly predicted the number of renal vessels in 27 of 28 cases (96%), including all 3 cases of left retroaortic renal vein. Also, 3DMRA correctly predicted the presence or absence of a crossing vessel in 10 of 12 cases (83%) of laparoscopic pyeloplasty. The imaging study also correctly predicted the number of hilar vessels in all five cases of radical nephrectomy, all three cases of partial nephrectomy, and both cases of other renal operations. Overall, 3D-MRA correctly defined the renal hilar anatomy in 48 of 50 patients, for an overall accuracy of 96%. CONCLUSIONS: Three-dimensional MRA findings correlate well (96%) with intraoperative findings in laparoscopic renal surgery. The imaging study provides exquisite vascular detail and is highly accurate, making it sufficient imaging prior to laparoscopic donor nephrectomy and useful for pyeloplasty and other complex renal operations. PMID- 15798419 TI - Retrieval and releasing capabilities of stone-basket designs in vitro. AB - BACKGROUND AND PURPOSE: A serious problem during calculus retrieval is the inability to release a stone from the basket. We evaluated a variety of basket designs to address this issue in two in-vitro models. MATERIALS AND METHODS: Four baskets--a modified, flat-wire, a double-helical, and two tipless Nitinol models (a passive and an articulating type)--were evaluated. The ability to retrieve and release beads of 4, 6, 8, 10, 12, and 14 mm diameter was compared in both a ureteral and a caliceal model. RESULTS: In the ureteral model, all baskets were successful in retrieving all sizes of beads as long as the bead was smaller than the measured basket diameter. Only the articulating (all sizes) and larger non articulating Nitinol models were able to release each of the bead sizes once engaged. The double-helical basket (larger size only) was able to release only the 4- and 6-mm beads after retrieval. For the 10- and 12-mm beads, the respective articulating Nitinol baskets could liberate the bead only when the articulating mechanism was used. In the caliceal model, the flat-wire and double helical baskets were unsuccessful in all attempts at retrieval. The larger non articulating Nitinol model was successful for beads >8 mm but had difficulty retrieving those smaller. Only the articulating Nitinol models were able to engage and release each of the beads. CONCLUSION: From these in-vitro studies, it is apparent that the Nitinol basket designs have the best retrieval capabilities. The ability to articulate the basket improves release of a stone once engaged. PMID- 15798421 TI - Comparison of a novel endoscope holder and traditional camera assistant for laparoscopic simple nephrectomy in a porcine model. AB - PURPOSE: To establish the feasibility of single-surgeon laparoscopy with application of a novel endoscope-holder device and to compare this technique with traditional assistant-driven laparoscopic camera control. MATERIALS AND METHODS: Bilateral simple nephrectomies were performed in six pigs. On one side, the operating surgeon employed the "scope holder" and performed the surgery without a laparoscopic assistant. On the contralateral side, an experienced camera operator was responsible for control of the laparoscopic field of vision in the traditional manner. The time required for hilar ligation and complete renal mobilization was documented. Pigs were sacrificed immediately after the procedure. RESULTS: The mean operative times for scope-holder and camera person assisted nephrectomy were 20.7 minutes and 19.3 minutes, respectively. The time to hilar ligation in the scope-holder and camera-operator-assisted cohorts was 13 and 14.5 minutes, respectively. There were no significant differences in operative times or blood loss in the two groups. The operative surgeon perceived some increase in shoulder and neck pain with use of the scope holder. CONCLUSIONS: This novel device provides a means for the operative surgeon to safely perform a laparoscopic nephrectomy alone without significantly increasing operative time or morbidity. PMID- 15798422 TI - Does prolonged pneumoperitoneum affect oxidative stress compared with open surgical procedures? AB - BACKGROUND AND PURPOSE: As laparoscopic surgery has evolved, it has become part of the urologic surgical armamentarium and is now used to perform more complex procedures. Carbon dioxide, used to create pneumoperitoneum, produces physiologic changes in various organs, including the kidneys. Such changes are associated with altered redox status because of the release of free radicals and changes in oxidative stress signals. It is unknown whether prolonged pneumoperitoneum is associated with an increase in oxidative stress compared with open surgery. The objective of this study was to compare oxidative stress in patients undergoing urologic laparoscopic and open operations. PATIENTS AND METHODS: Urine samples were obtained immediately preoperatively, immediately postoperatively, and at 6 and 18 hours after surgery from 10 patients who underwent urologic laparoscopic surgery and 10 patients who underwent open surgery. Concentrations of the oxidative stress marker isoprostane (8- iso-prostaglandin F2a) were measured, and the results were analyzed with respect to clinical factors associated with the type of surgery. RESULTS: Urinary isoprostane concentrations (mean +/- SEM) in the laparoscopic and open groups showed an increase immediately after surgery to 189.0 +/- 64.2% and 141.1 +/- 45.8% of the preoperative values, respectively. A decrease in isoprostane was subsequently observed in both groups at 6 hours postoperatively, with preoperative values restored at 18 hours postoperatively (126.3 +/- 19.7% and 89.5 +/- 55.9% at 6 and 18 hours, respectively, in the laparoscopic group and 130.7 +/- 41.6% and 88.7 +/- 20.4% at 6 and 18 hours, respectively, in the open-surgery group). Although in both groups the peak PGF 2a concentration was observed immediately (0 hours) postoperatively, no significant differences were observed between the groups at 0, 6, and 18 hours. In the laparoscopic-surgery group, the mean increase tended to be higher and the decrease to be less prolonged than in the open-surgery group. CONCLUSION: Oxidative stress, as measured by urinary 8-iso-prostaglandin F2a, is produced by both laparoscopic and open urologic surgery. The findings of our nonrandomized study suggest a pattern of increased oxidative stress postoperatively with either type of surgery, with subsequent return almost to preoperative levels. Prolonged laparoscopic operative time did not affect oxidative stress levels. PMID- 15798423 TI - Laparoscopic ureterocalicostomy: development of a technique simplified by application of Nitinol clips and a wet monopolar electrosurgery device. AB - PURPOSE: We developed a technique for laparoscopic ureterocalicostomy with the use of intracorporeal suturing and subsequently simplified the technique by application of experimental Nitinol clips. MATERIALS AND METHODS: We performed laparoscopic ureterocalicostomy on 16 domestic swine divided into four groups of four animals each. The kidney was exposed laparoscopically, and the renal artery was atraumatically clamped. The lower pole of the kidney was amputated to expose a lower-pole calix, and hemostasis of the cut renal surface was obtained with a wet monopolar electrosurgical device (Floating Ball device [FB]; TissueLink, Dover, NH). Anastomosis of the ureter to the lower-pole calix was performed over a guidewire using 3-0 Vicryl suture in group 1 and Nitinol clips in group 3. A double-J ureteral stent was then deployed retrograde under fluoroscopic guidance. In addition, we evaluated the use of fibrin glue as a sealant over the sutured or clipped anastomotic site (groups 2 and 4, respectively). Ureteral stents were removed after 3 weeks, and the animals were evaluated and sacrificed after an additional 3 weeks. RESULTS: Laparoscopic ureterocalicostomy was completed in all 16 animals. In each case, excellent renal parenchymal hemostasis was obtained with the FB device, with a mean hemostasis time of 4.1 minutes. The mean anastomotic time with standard suture reconstruction was 37.1 +/- 5.4 minutes, while the anastomotic time with the Nitinol clips was 29.0 +/- 8.0 minutes (P = 0.0339). Retrograde pyelograms in groups 1 and 3 (no fibrin glue) showed a patent anastomosis with no hydronephrosis in three of the four animals in each group. One animal in group 1 and one animal in group 3 developed large urinomas secondary to anastomotic failure. The animals that received fibrin glue over the anastomotic site (groups 2 and 4) all showed narrowed anastomoses with severe hydronephrosis. CONCLUSIONS: With available instrumentation, laparoscopic ureterocalicostomy is technically feasible. Nitinol clip technology significantly reduces collecting-system reconstruction time. Application of fibrin glue as a urinary tract sealant resulted in an unexpected adverse outcome. PMID- 15798425 TI - High-powered holmium-laser ablation: comparison of setting effectiveness using tissue-surrogate model. AB - BACKGROUND AND PURPOSE: The 100-W holmium laser, in conjunction with a sidefiring 550-microm fiber, can ablate prostate glands as large as 40 g to relieve lower urinary-tract symptoms. We evaluated the effect of various setting combinations on ablation efficiency using beef kidney as a tissue-surrogate model. MATERIALS AND METHODS: Beef kidney specimens (mean weight 44.8 +/- 3.1 g) were secured in a cylinder, which was submerged in a water-filled tank through which a 27F resectoscope and 550-microm sidefiring fiber were positioned. Four energy/frequency combinations were tested, with each used to treat 10 kidney specimens. The difference between the mean pretreatment and post-treatment weights of each treatment group were compared statistically with Student's t test. RESULTS: The largest mean weight difference after treatment (8.94 +/- 2.38 g) was achieved using 3.2 J and 25 Hz. This mass reduction was significantly greater than that of all other combinations except 2.5 J and 40 Hz. CONCLUSIONS: Use of the 3.2 J and 25 Hz setting combination resulted in the greatest amount of ablation in this tissue-surrogate model, suggesting that maximal energy settings may provide an advantage in tissue vaporization using the 100-W holmium laser. Clinical assessment must be performed to substantiate these findings. PMID- 15798426 TI - Geometry and temperature distribution during radiofrequency tissue ablation: an experimental ex vivo model. AB - PURPOSE: To investigate the temperature distribution and geometry of lesions created by radiofrequency (RF) tissue ablation. MATERIALS AND METHODS: We developed an ex-vivo thermal model based on the ability of egg whites to coagulate at 62 degrees to 65 degrees C. This property allows us to observe and record the formation of "lesions" created by different commercially available RF generators and probes. RESULTS: The lesions created by the Radionics Cool-Tip RF system were mostly cylindrical or barrel-shaped, while RITA StarBurst XL and StarBurst XLi electrodes produced cone-like or mushroom-shaped lesions. The time required to achieve maximum lesion size was between 3 and 5 minutes in most trials. The formation of lesions stopped when thermodynamic equilibrium was achieved. CONCLUSIONS: The geometry of lesions does not correlate precisely with manufacturer reports, which describe spherical lesions for both RF systems. The 12-minute treatment cycle recommended by company's algorithm may not be necessary because lesion formation was completed and thermodynamic equilibrium was reached significantly earlier. We believe the results of our study may be useful in better understanding the RF ablation process, better planning, and improvement of clinical outcome. PMID- 15798424 TI - The Heilbronn laparoscopic training program for laparoscopic suturing: concept and validation. AB - BACKGROUND AND PURPOSE: More than a decade after the first description of laparoscopic nephrectomy, an increasing number of laparoscopic procedures are being performed worldwide. Nevertheless, there still exists a significant lack of standardized training programs to teach ablative and, most important, reconstructive laparoscopic operations (i.e., pyeloplasty, radical prostatectomy). We evaluated and validated a new standardized step-by-step program to improve laparoscopic skills and enable trainees not experienced in laparoscopy to perform a urethrovesical anastomosis. MATERIALS AND METHODS: In an inanimate model (pelvic trainer) with defined trocar positions, the 10 participants were exposed to six reconstructive exercises. The steps consist of improvement of hand-eye coordination (two-row metal-pin model; step I), linear and curved suturing with changing angles of the needle (chicken leg and catheter model; steps II-V), and performance of an anastomosis in a porcine bladder (step VI). Times of 3, 15, 15, 10, 20, and 30 minutes for steps I, II, III, IV,V, and VI, respectively, were defined as the goal before proceeding to the next stage. The time required to succeed in each step and the increase in the speed of suturing and knotting activities were analyzed with the Wilcoxon signed-rank test. RESULTS: After a mean of 40 hours of training, all participants were able to perform all steps within the specified times and complete an accurate urethrovesical anastomosis in 30 minutes. The time required to succeed before and after training showed a significant decrease (P < 0.05). Continual training in reconstructive procedures decreased the time needed for suturing activities by between 66.3% and 72.2%. The time needed for the knotting activities decreased by between 34.3% and 38.3%. CONCLUSIONS: Our program enabled participants not experienced in laparoscopy to increase reproducible performance in reconstructive laparoscopy. These results indicate that the challenging parts of reconstructive laparoscopy such as intracorporeal suturing can be taught using a standardized concept. This experience could be incorporated easily by every department developing a laparoscopic training program. PMID- 15798427 TI - Contrast-enhanced ultrasound evaluation of radiofrequency ablation of the kidney: reliable imaging of the thermolesion. AB - BACKGROUND AND PURPOSE: Radiofrequency ablation (RFA) is limited by the inability to monitor progression of the thermal lesion. Contrast-enhanced ultrasonography (CEUS) imaging has considerable potential as a monitoring modality for RFA. We report our experience using CEUS to evaluate the lesion produced by RFA in a porcine model. MATERIALS AND METHODS: Five pigs underwent laparoscopic RFA twice, spaced by a 1-week interval. Post- RFA ultrasound imaging was performed transcutaneously immediately after ablation. The kidneys were assessed for a contrast void corresponding to the ablated tissue. The kidneys were then harvested and the gross RFA lesions measured to compare lesion size with that measured using CEUS. RESULTS: A clear lesion was identified at the site of each RFA application. As measured by CEUS, the acute lesions averaged 1.8 +/- 0.4, 1.7 +/- 0.4, and 1.8 +/- 0.3 cm in length, height, and width, respectively, compared with 2.0 +/- 0.5, 1.8 +/- 0.4, and 1.9 +/- 0.3 cm, respectively, by measurement in the gross specimen (P = 0.33, 0.13, and 0.44, respectively). At 1 week, the CEUS-measured dimensions of the lesions were 2.7 +/- 0.5, 2.6 +/- 0.6, and 2.6 +/ 0.6 cm, and the gross measurements of the lesion were 2.7 +/- 0.4, 2.7 +/- 0.9, and 2.6 +/- 0.4 cm (P = 0.75, 0.92, and 0.40, respectively). CONCLUSIONS: Contrast-enhanced ultrasonography appears to be an accurate modality for immediate monitoring of RFA defects. Further study is necessary to assess the clinical utility of CEUS for monitoring RFA of small renal lesions. PMID- 15798428 TI - LiteratureWatch, July-December 2004. PMID- 15798432 TI - Light microscopic, immunohistochemical, and ultrastructural alterations in patients with melasma. AB - Despite new technologies, few studies have assessed the histologic alterations in patients with melasma. Using current technologies, the present study was designed to re-evaluate the light microscopic, immunohistochemical, and ultrastructural changes of the hyperpigmented and adjacent normal skin of patients with melasma. Twenty-one patients were included in this study. Two millimeter punch biopsies were taken from the hyperpigmented and adjacent normal skin of the face. The integrity of the epidermis and dermis was assessed by light microscopy, computer assisted image analysis, immunohistochemistry, and electron microscopy. Stains included hematoxylin-eosin and Fontana-Masson for melanin detection. Immunostaining was performed using Mel-5 antibody and CD1a antibody as markers for melanin and Langerhans cells, respectively. However, mild lymphohistiocytic infiltrates were present in 75% of the hyperpigmented areas. The areas of hyperpigmentation showed increased deposition of melanin in the epidermis and dermis of all cases. There was a statistically significant increase in the content of epidermal melanin. There were no quantitative increases in melanocytes in the hyperpigmented areas of skin. However, the melanocytes in the hyperpigmented areas were larger, intensely stained cells with very prominent dendrites. Electron microscopy revealed more melanosomes in keratinocytes, melanocytes, and dendrites in the involved skin in comparison to the uninvolved skin. The results of this study suggest that melasma is a consequence of specific hyperfunctional melanocytes that cause excessive melanin deposition in the epidermis and dermis. PMID- 15798431 TI - Androgen receptor expression helps to differentiate basal cell carcinoma from benign trichoblastic tumors. AB - Histologic differentiation between basal cell carcinoma and benign trichoblastic neoplasms such as trichoepithelioma and trichoblastoma can be difficult on small biopsies. Therefore, several attempts have been made to identify immunohistochemical differences between these entities. Recent studies have shown androgen receptor expression in a number of mature epithelial structures in the skin and in epithelial neoplasms including basal cell carcinoma. In contrast, androgen receptor expression was absent in mature hair follicles or the few trichogenic neoplasms studied to date. These findings suggested that androgen receptor expression might be a useful adjunct in the histologic differential diagnosis between basal cell carcinoma and benign trichoblastic neoplasms. Therefore, we performed immunohistochemical analysis of androgen receptor expression in 32 basal cell carcinomas and 10 benign trichoblastic tumors (6 trichoepitheliomas and 4 trichoblastomas). In our study, at least focal expression of androgen receptor was detected in 78% of basal cell carcinomas. None of the trichoblastic tumors showed any androgen receptor immunoreactivity. These results confirm the lack of expression of androgen receptor in benign trichoblastic neoplasms and indicate that androgen receptor expression by tumor cells points to basal cell carcinoma as the most likely diagnosis. PMID- 15798434 TI - Melanocytic nevi of the auricular region: histologic characteristics and diagnostic difficulties. AB - We examined auricular melanocytic nevi to evaluate their architectural and cytologic features. A retrospective analysis of 21 auricular melanocytic nevi was conducted during 3 years. The nevi were evaluated for cytologic atypia, architectural disorder, location of epidermal nests, pagetoid spread, growth pattern (symmetry versus asymmetry), demarcation of lateral borders, and host response (lymphocytic infiltrate and/or lamellar fibroplasia). Eleven cases (52.4%) measured at least 6 mm in greatest dimension and 6 cases (28.6%) were asymmetric. Fourteen cases (66.7%) were ill demarcated. Pagetoid spread was present in 12 cases (57.1%). Ten cases (47.6%) showed moderate to severe cytologic atypia. Nucleoli were prominent in 9 cases (42.8%). None of the cases showed mitoses or apoptotic melanocytes. Auricular melanocytic nevi, like those occurring in the acral, flexural, and genital areas, may exhibit some histologic features commonly found in melanomas. Careful histologic interpretation of these lesions is recommended. PMID- 15798433 TI - Hidradenoma papilliferum with oxyphilic metaplasia: a clinicopathological study of 18 cases, including detection of human papillomavirus. AB - Reported here are 18 cases of hidradenoma papilliferum with oxyphilic metaplasia. All patients were women ranging in age from 29 to 74 years. Each presented clinically with a small, solitary tumor in the anogenital region. Microscopically, in addition to classic histopathological features, in every case there was oxyphilic metaplasia of the constituent epithelial cells. This finding could be likened to apocrine metaplasia, a term used in breast pathology. Other histopathological findings observed in this series, analogous to benign breast disease, included sclerosing adenosis-like changes, atypical apocrine adenosis like changes, changes corresponding to usual ductal epithelial hyperplasia, epitheliomatosis with a streaming growth pattern, lamprocyte-like changes, clear cell change of the myoepithelium, foamy histiocyte reaction, and stromal fibrosis. Immunohistochemistry inferred that in the majority of cases oxyphilic metaplasia resulted from more lysosomes, whereas numerous mitochondria were detected in only 3 cases. Using 2 different PCR methods we identified HPV in 4 of 15 cases of hidradenoma with oxyphilic metaplasia. In addition, HPV was detected in 3 of 16 conventional papillary hidradenomas used as a control group. The following HPV types were identified: 16, 31, 33, 53, and 56. The last type was found in 5 cases. More than one HPV type from a single lesion was seen in 5 cases. Our observations are consistent with previous publications noting similarities between tumors of the breast and sweat glands. Oxyphilic metaplasia, areas with solid growth, and changes simulating atypical apocrine adenosis are rare and poorly recognized in hidradenoma papilliferum and may cause diagnostic difficulties; in our cases several submitting pathologists suspected malignancy. A causal role for HPV in hidradenoma papilliferum cannot be confirmed from our results, as the detection rate is too low. The exact role of the HPV in etiology and pathogenesis of this neoplasm has yet to be determined. PMID- 15798435 TI - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is expressed in normal skin and cutaneous inflammatory diseases, but not in chronically UV exposed skin and non-melanoma skin cancer. AB - Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) is a member of the tumor necrosis factor family that preferentially induces apoptosis in transformed but not normal cells and that is constitutively expressed in many organs including the skin. In addition to its therapeutic potential, TRAIL might act as a natural guardian eliminating transformed cells at an early stage. Ultraviolet (UV) radiation is not only a potent carcinogen because of its mutagenic effects but also because of its capacity to paralyze natural protection mechanisms, including the tumor suppressor gene p53. Therefore, we studied the effect of UV exposure on the expression of TRAIL in the skin by immunohistochemical analysis. TRAIL and its receptors TRAIL-R1 and TRAIL-R4 were constitutively expressed in normal epidermis and not altered in a variety of inflammatory dermatoses including those associated with interface dermatitis. TRAIL was not altered in biopsies of acute sunburn, polymorphic light eruption, and photoprovocation testing, indicating that acute UV exposure does not affect TRAIL expression. No differences were observed in UV-protected and chronically UV exposed skin samples of younger adults. In contrast, TRAIL was significantly reduced in chronically UV-exposed skin of elderly individuals. In addition, TRAIL expression was reduced in actinic keratoses and Bowen disease and almost completely lost in basal cell and squamous cell carcinomas. In contrast, keratoacanthomas did not reveal any alterations in TRAIL expression. Taken together, these data indicate that chronic UV exposure in elderly patients results in the loss of TRAIL expression, which might contribute to the increased risk of skin cancer in this population. Down-regulation of TRAIL might represent another example of a natural protection mechanism that is eliminated by chronic UV exposure. PMID- 15798436 TI - A morphological study of ectopic plantar nail. AB - Ectopic nails are rare. Four adult patients presented to us, affected by true ectopic nails of the foot, sole, and toes, occurring bilaterally in one case. Full documentation of these anomalies was collected. The aim of this study was to better describe the morphologic features of ectopic plantar nails by using, besides histologic evaluation, transmission and scanning electron microscopy. New morphologic criteria were identified to determine if ectopic plantar nail differed from normal and ectopic hand nails. Morphologically, ectopic plantar nails are smaller than normal ones, but still form cuticles, nail grooves and nail folds, and periungual wrinkles. Furthermore, they form with a flat angle between the nail matrix and proximal nail fold, and have a nail plate interrupted by several transverse nails grooves. PMID- 15798437 TI - Malignant melanoma with osteoclast-like giant cells: an unusual host response: immunohistochemical and ultrastructural study of three cases and literature review. AB - Melanomas with unusual histologic features are very rarely reported in the literature and demonstrate the diversity of melanocytic expression. Three cases of malignant melanoma with osteoclast-like giant cells are reported. Two cases showed undifferentiated malignant cells without melanin pigment and one showed spindled cell morphology. Immunohistochemistry showed that the osteoclast- like giant cells expressed CD68, but not melanocytic markers (HMB45, Melan-A, and S100). Ultrastructural analysis further supports that these cells are reactive histiocytes rather than transformed malignant cells. This suggests they represent an unusual host response, similar to those rarely observed in other neoplasms. Awareness of this entity is important to avoid misdiagnosis of melanoma as a histiocytic tumor. Since only few cases have been reported, greater recognition and documentation may help to evaluate the prognosis of such cases with unusual morphology. PMID- 15798438 TI - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephaloapthy (CADASIL): a hereditary cerebrovascular disease, which can be diagnosed by skin biopsy electron microscopy. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited cerebrovascular disease characterized by recurrent subcortical ischemic strokes starting in the third or fourth decade as a result of mutations in the Notch3 gene. Granular osmiophilic material (GOM) deposition around the vascular smooth muscle cells is a specific feature and electron microscopic observations of skin biopsies are useful for this diagnosis. A 39-year-old female with dizziness, abnormal visual fields, and hemiplegia, and a 42-year-old male with tinnitus and dizziness, were suspected of suffering from CADASIL based on MRI findings. Both cases were shown to have characteristic deposits of GOM, 200 to 800 nm in diameter, around the vascular smooth muscle cells of small arteries in the deep dermis, and thus the diagnoses of CADASIL were made, although there was no family history of cerebrovascular disorders or dementia. Dermatologists should be aware of these ultra-structural findings because this disease may occur sporadically and might be more common than initially thought. PMID- 15798439 TI - Reactive syringofibroadenomatous hyperplasia in peristomal skin with formation of hybrid epidermal-colonic mucosa glandular structures, intraepidermal areas of sebaceous differentiation, induction of hair follicles, and features of human papillomavirus infection: a diagnostic pitfall. AB - We report a case of reactive syringofibroadenomatous hyperplasia in peristomal skin. The patient was a 62-year-old woman who had undergone abdominoperineal resection of the rectum for rectal adenocarcinoma with subsequent colostomy 2 years earlier. Clinically, a nodule and small, whitish, warty lesions developed at the outer margin of the stoma extending onto the adjacent skin. Following a clinical suspicion of adenocarcinoma, recurrent at the colostomy site, a 5 x 4 x 3-cm excision of the peristomal skin and the affected portion of the stoma was performed and submitted for histologic examination. The biopsy revealed a peculiar composite lesion of reactive syringofibroadenomatous hyperplasia and the excised part of the stoma. Several unusual histopathological features were detected in the syringofibroadenomatous part of the lesion such as the formation of plentiful hybrid epidermal-colonic mucosa glandular structures, intraepidermal areas of sebaceous differentiation, koilocytic changes, induction of rudimentary hair follicles, and intradermal mucinous lakes. The cellular composition of the glandular structures was mainly similar to that seen in a normal colonic mucosa epithelium. They also contained occasional Paneth cells. Being located at a distance from the stoma, these accentuated colonic mucosa epithelial glands reaching the epidermis may be a diagnostic pitfall prompting the consideration of adenocarcinoma involving the stoma. The rudimentary follicles and sebaceous differentiation were probably induced by an altered stroma and/or human papillomavirus (HPV): HPV, type 36 was identified by PCR using consensus primers followed by sequencing of the PCR products. PMID- 15798440 TI - Pseudoangiosarcomatous squamous cell carcinoma of skin arising adjacent to decubitus ulcers. AB - Pseudoangiosarcomatous, or pseudovascular, squamous cell carcinoma of skin is an unusual form of acantholytic (adenoid, pseudoglandular) squamous cell carcinoma that mimics the histolopathologic appearance of angiosarcoma. We report a case of pseudoangiosarcomatous squamous cell carcinoma arising adjacent to decubitus ulcers. The histopathologic examination of a wedge biopsy specimen revealed infiltrative cords of neoplastic cells that formed interanastomosing channels imitating angiosarcoma. Immunohistochemical staining was negative for the endothelial markers (CD31, CD34, and factor VIII-related antigen) and positive using cytokeratin antibodies (AE1/AE3 and 34 betaE12). Because of metastatic disease, palliative measures were undertaken and the patient died four months later. To our knowledge, our patient is the first with pseudoangiosarcomatous squamous cell carcinoma of skin developing within decubitus ulcer. PMID- 15798441 TI - Gastric mucosa in a bronchogenic cutaneous cyst in a child: case report and review of literature. AB - We report a case of cutaneous bronchogenic cyst, partially lined by gastric mucosa of antral type in a 9-year-old boy. This is, to the best of our knowledge, the first report of gastric mucosa in bronchogenic cutaneous cyst in the literature. PMID- 15798442 TI - Fibroepithelioma of pinkus is a fenestrated trichoblastoma. AB - Pinkus described "pre-malignant fibroepithelioma" as a proliferation that gave rise to many tiny basal cell carcinomas within each lesion. Later authors have generally considered it to be an unusual variant of basal cell carcinoma (BCC). The delineation of trichoblastoma as the general term for the benign counterpart of BCC raises the possibility that the fibroepithelioma of Pinkus (FEP) would be better classified under that rubric. To address this subject, we examined the records of 114 patients with FEP for body site, age and sex distribution, and sections from 75 lesions. All FEP examined show a blunt interface with the underlying dermis (where one could be seen), differentiation toward follicular bulbs and papillae, and large areas of cellular stroma. FEP has a slight female preponderance in contrast to BCC, which is more common in males. Unlike the common types of BCC, FEP has an overwhelming predilection for the trunk and extremities, and only 5% of tumors are set in a dermis with significant amounts of solar elastosis. Next, FEP, BCC, and FEP with BCC-like areas were stained with MIB-1 (to assess proliferation), p53 (an oncogene product), and CK20 (a Merkel cell marker) antisera. FEP shows a low level of staining for p53 and MIB-1, in contrast to conventional BCCs that over-express these markers. FEP also shows retention of Merkel cells, a characteristic of benign neoplasms with follicular germinative differentiation but not in general of BCC. The BCC-like areas in some FEP tumors reflect these staining tendencies with less striking differences. Given the contrast between FEP and BCC with respect to site of occurrence, relationship to sun damage, histopathologic features, and immunohistochemical studies, it appears that FEP more closely resembles trichoblastoma than BCC. PMID- 15798443 TI - Fibroepithelial tumor of pinkus is trichoblastic (Basal-cell) carcinoma. PMID- 15798444 TI - Staging of melanoma: a critique in historical perspective: part I. AB - During the last half of the 20th century until the present day, general pathologists have been obsessed by staging melanoma. That the method employed for achieving that desideratum, which had as its animating force a longing to determine prognosis for an individual patient with that malignant neoplasm, failed miserably can be inferred from the fact that successive efforts every few years by teams of supposed authorities, such as those of the American Joint Committee on Cancer (AJCC), resulted in a new system for staging that bore little resemblance to the one immediately before it or the one that was to come after it. In short, the various attempts to stage melanoma were nothing more than witchcraft and sorcery in which pathologists sought to be diviners, seers, and prophets about prognosis, rather than the diagnosticians that they were trained to be and should be. The job of a pathologist is to render an accurate diagnosis (which, in itself, has implications prognostic for a patient, eg, "benign" or "malignant"), not to speculate about prognosis. To guess about how a patient with melanoma will fare, the system for staging being nothing other than pure conjecture, not only is unscientific, it is unethical. PMID- 15798445 TI - Staging of melanoma: a critique of the most recent (2002) system proposed by the American Joint Committee on Cancer: part II. AB - The most recent effort in 2002 by the American Joint Committee on Cancer for staging melanoma is not a job more effective than the first endeavor of that Committee in 1962; conjecture, speculation, and supposition reign. PMID- 15798446 TI - A baffling basaloid blain. AB - Cutaneous metastases may present in a variety of histologic guises simulating more common primary tumors. We present the clinicopathologic features of a case masquerading as basal cell carcinoma. Epidermotropism, vascular invasion, and the absence of stromal retraction around tumor nests prompted further investigation. Clinical history and directed immunohistochemical staining ultimately revealed the diagnosis of cutaneous metastasis of transitional cell bladder carcinoma (TCC). PMID- 15798447 TI - Penoscrotal extramammary Paget's disease in a patient with rectal cancer: double primary adenocarcinomas differentiated by immunoperoxidase staining. PMID- 15798448 TI - Mucocele of the neck. PMID- 15798449 TI - Cutaneous epithelioid angiomatous nodule of the external ear. PMID- 15798450 TI - Cutaneous keratocyst arising independently of the nevoid basal cell carcinoma syndrome. PMID- 15798451 TI - CD10, a useful marker for atypical fibroxanthomas. PMID- 15798452 TI - Cutaneous lymphocytic infiltrates: let's get real. PMID- 15798453 TI - Alcohol interventions for trauma patients treated in emergency departments and hospitals: a cost benefit analysis. AB - OBJECTIVE: To determine if brief alcohol interventions in trauma centers reduce health care costs. SUMMARY BACKGROUND DATA: Alcohol-use disorders are the leading cause of injury. Brief interventions in trauma patients reduce subsequent alcohol intake and injury recidivism but have not yet been widely implemented. METHODS: This was a cost-benefit analysis. The study population consisted of injured patients treated in an emergency department or admitted to a hospital. The analysis was restricted to direct injury-related medical costs only so that it would be most meaningful to hospitals, insurers, and government agencies responsible for health care costs. Underlying assumptions used to arrive at future benefits, including costs, injury rates, and intervention effectiveness, were derived from published nationwide databases, epidemiologic, and clinical trial data. Model parameters were examined with 1-way sensitivity analyses, and the cost-benefit ratio was calculated. Monte Carlo analysis was used to determine the strategy-selection confidence intervals. RESULTS: An estimated 27% of all injured adult patients are candidates for a brief alcohol intervention. The net cost savings of the intervention was 89 US dollars per patient screened, or 330 US dollars for each patient offered an intervention. The benefit in reduced health expenditures resulted in savings of 3.81 US dollars for every 1.00 US dollar spent on screening and intervention. This finding was robust to various assumptions regarding probability of accepting an intervention, cost of screening and intervention, and risk of injury recidivism. Monte Carlo simulations found that offering a brief intervention would save health care costs in 91.5% of simulated runs. If interventions were routinely offered to eligible injured adult patients nationwide, the potential net savings could approach 1.82 billion US dollars annually. CONCLUSIONS: Screening and brief intervention for alcohol problems in trauma patients is cost-effective and should be routinely implemented. PMID- 15798454 TI - Alcohol interventions for trauma patients treated in emergency departments: can we afford not to intervene? PMID- 15798456 TI - Can we be sure polypropylene mesh causes infertility? PMID- 15798455 TI - Herniorrhaphy with polypropylene mesh causing inguinal vasal obstruction: a preventable cause of obstructive azoospermia. AB - OBJECTIVE: To report a multiinstitutional experience of men presenting with infertility secondary to inguinal hernia repair using polypropylene mesh. SUMMARY BACKGROUND DATA: An estimated 80% of inguinal hernia operations involve placement of a knitted polypropylene mesh to form a "tension-free" herniorrhaphy. The prosthetic mesh induces a chronic foreign-body fibroblastic response creating scar tissue that imparts strength to the floor and leads to fewer recurrences. However, little is known about the long-term effects of the polypropylene mesh on the vas deferens, especially with regard to fertility. METHODS: Eight institutions in the United States reported a total of 14 cases of azoospermia secondary to inguinal vasal obstruction related to previous polypropylene mesh herniorrhaphy. Patient characteristics and operative findings were forwarded to 1 center for tabulation of data. RESULTS: Mean patient age was 35.5 years with an average duration of infertility of 1.8 years. Mean number of years between urologic evaluation and herniorrhaphy was 6.3 years. Types of inguinal hernia repair previously performed were: open (10), laparoscopic (2), or both (2). Nine patients had bilateral obstruction and 5 patients had unilateral obstruction with contralateral testicular atrophy or epididymal obstruction. Surgical exploration revealed a dense fibroblastic response encompassing the polypropylene mesh with either trapped or obliterated vas in all patients. Surgical reconstruction was performed in 8 of 14 men (57%). CONCLUSION: Reconstruction to restore fertility can be difficult secondary to fibrotic reaction. Before undergoing polypropylene mesh herniorrhaphy, men, especially of young reproductive age or with a solitary testicle, need to be carefully advised of potential obstruction and compromise to future fertility. PMID- 15798457 TI - Systematic review of surgical options for idiopathic megarectum and megacolon. AB - OBJECTIVE: A subgroup of patients with intractable constipation has persistent dilatation of the bowel, which in the absence of an organic cause is termed idiopathic megabowel (IMB). The aim of this systematic review was to evaluate the published outcome data of surgical procedures for IMB in adults. METHODS: Electronic searches of the MEDLINE (PubMed) database, Cochrane Library, EMBase, and Science Citation Index were performed. Only peer-reviewed articles of surgery for IMB published in the English language were evaluated. Studies of all surgical procedures were included, providing they were performed on 3 or more patients, and overall success rates were documented. Studies were critically appraised in terms of design and methodology, inclusion criteria, success, mortality and morbidity rates, and functional outcomes. RESULTS: A total of 27 suitable studies were identified, all evidence was low quality obtained from case series, and there were no comparative studies. The studies involved small numbers of patients (median 12, range 3-50), without long-term follow-up (median 3 years, range 0.5 7). Inclusion of subjects, methods of data acquisition, and reporting of outcomes were extremely variable. Subtotal colectomy was successful in 71.1% (0%-100%) but was associated with significant morbidity related to bowel obstruction (14.5%, range 0%-29%). Segmental resection was successful in 48.4% (12.5%-100%), and recurrent symptoms were common (23.8%). Rectal procedures achieved a successful outcome in 71% to 87% of patients. Proctectomy, the Duhamel, and pull-through procedures were associated with significant mortality (3%-25%) and morbidity (6% 29%). Vertical reduction rectoplasty (VRR) offered promising short-term success (83%). Pelvic-floor procedures were associated with poor outcomes. A stoma provided a safe alternative but was only effective in 65% of cases. CONCLUSIONS: Outcome data of surgery for IMB must be interpreted with extreme caution due to limitations of included studies. Recommendations based on firm evidence cannot be given, although colectomy appears to be the optimum procedure in patients with a nondilated rectum, restorative proctocolectomy the most suitable in those with dilatation of the colon and rectum, and VRR in those patients with dilatation confined to the rectum. Appropriately designed studies are required to make valid comparisons of the different procedures available. PMID- 15798458 TI - A comparison of ductoscopy-guided and conventional surgical excision in women with spontaneous nipple discharge. AB - INTRODUCTION: Radiologic imaging is routinely used to evaluate women with spontaneous nipple discharge (SND), but definitive diagnosis is usually only achieved by surgical terminal duct excision (TDE). Ductoscopy has been reported to result in improved localization of intraductal lesions and may avoid surgery in women with endoscopically normal ducts. MATERIALS AND METHODS: We conducted a retrospective review of the records of 117 consecutive women who underwent ductoscopy to guide ductal excision (scope-DE) or received conventional TDE without ductoscopy. Two women had atypical ductal lavage cytology and the remainder presented with SND from 1 or more duct. Preoperative evaluation included radiologic imaging as clinically indicated. RESULTS: Fifty-nine women underwent scope-DE, and 58 underwent conventional TDE. There were no significant differences in age, race, discharge characteristics, or radiologic findings. The proportion of women with intraductal neoplasia was slightly greater in the group undergoing scope-DE (88% vs. 81%, P = 0.2). In the conventional TDE group, 8.5% were found to have atypical hyperplasia or duct carcinoma in situ compared with 18.6% in the scope-DE group. In the ductoscopy group, 22 of 59 (37.3%) had lesions >5 cm from the nipple, compared with 1 of 17 women for whom distance of the lesion from the nipple was known in the conventional group (P = 0.02). Of the ductoscopy-detected cancers, 5 of 6 had no symptoms other than SND, whereas 1 of the 4 malignancies in the conventional group presented as SND alone. DISCUSSIONS: Ductoscopy identifies intraductal lesions in a high proportion of women with SND, and it may contribute to more accurate resection of these. A prospective study is required to obtain an unbiased assessment of these possible advantages. PMID- 15798459 TI - Complete versus selective portal triad clamping for minor liver resections: a prospective randomized trial. AB - OBJECTIVE: To evaluate the feasibility, safety, efficacy, amount of hemorrhage, postoperative complications, and ischemic injury of selective clamping in patients undergoing minor liver resections. SUMMARY BACKGROUND DATA: Inflow occlusion can reduce blood loss during hepatectomy. However, Pringle maneuver produces ischemic injury to the remaining liver. Selective hemihepatic vascular occlusion technique can reduce the severity of visceral congestion and total liver ischemia. PATIENTS AND METHODS: Eighty patients undergoing minor hepatic resection were randomly assigned to complete clamping (CC) or selective clamping (SC). Hemodynamic parameters, including portal pressure and the hepatic venous pressure gradient (HVPG), were evaluated. The amount of blood loss, measurements of liver enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST), and postoperative evolution were also recorded. RESULTS: No differences were observed in the amount of hemorrhage (671 +/- 533 mL versus 735 +/- 397 mL; P = 0.54) or the patients that required transfusion (10% versus 15%; P = 0.55). There were no differences on postoperative morbidity between groups (38% versus 29%; P = 0.38). Cirrhotic patients with CC had significantly higher ALT (7.7 +/- 4.6 versus 4.5 +/- 2.7 mukat/L, P = 0.01) and AST (10.2 +/- 8.7 versus 4.9 +/- 2.1 mukat/L; P = 0.03) values on the first postoperative day than SC. The multivariate analysis demonstrated that high central venous pressure, HVPG >10 mm Hg, and intraoperative blood loss were independent factors related to morbidity. CONCLUSIONS: Both techniques of clamping are equally effective and feasible for patients with normal liver and undergoing minor hepatectomies. However, in cirrhotic patients selective clamping induces less ischemic injury and should be recommended. Finally, even for minor hepatic resections, central venous pressure, HVPG, and intraoperative blood loss are factors related to morbidity and should be considered. PMID- 15798460 TI - Long-term follow-up of a randomized trial comparing the beger and frey procedures for patients suffering from chronic pancreatitis. AB - OBJECTIVE: To report on the long-term follow-up of a randomized clinical trial comparing pancreatic head resection according to Beger and limited pancreatic head excision combined with longitudinal pancreatico-jejunostomy according to Frey for surgical treatment of chronic pancreatitis. SUMMARY BACKGROUND DATA: Resection and drainage are the 2 basic surgical principles in surgical treatment of chronic pancreatitis. They are combined to various degrees by the classic duodenum preserving pancreatic head resection (Beger) and limited pancreatic head excision combined with longitudinal pancreatico-jejunostomy (Frey). These procedures have been evaluated in a randomized controlled trial by our group. Long-term follow up has not been reported so far. METHODS: Seventy-four patients suffering from chronic pancreatitis were initially allocated to DPHR (n = 38) or LE (n = 36). This postoperative follow-up included the following parameters: mortality, quality of life (QL), pain (validated pain score), and exocrine and endocrine function. RESULTS: Median follow-up was 104 months (72-144). Seven patients were not available for follow-up (Beger = 4; Frey = 3). There was no significant difference in late mortality (31% [8/26] versus 32% [8/25]). No significant differences were found regarding QL (global QL 66.7 [0-100] versus 58.35 [0-100]), pain score (11.25 [0-75] versus 11.25 [0-99.75]), exocrine (88% versus 78%) or endocrine insufficiency (56% versus 60%). CONCLUSIONS: After almost 9 years' long-term follow-up, there was no difference regarding mortality, quality of life, pain, or exocrine or endocrine insufficiency within the 2 groups. The decision which procedure to choose should be based on the surgeon's experience. PMID- 15798461 TI - Glutamine supplementation of parenteral nutrition does not improve intestinal permeability, nitrogen balance, or outcome in newborns and infants undergoing digestive-tract surgery: results from a double-blind, randomized, controlled trial. AB - OBJECTIVE: To assess the effect of isocaloric isonitrogenous parenteral glutamine supplementation on intestinal permeability and nitrogen loss in newborns and infants after major digestive-tract surgery. SUMMARY BACKGROUND DATA: Glutamine supplementation in critically ill and surgical adults may normalize intestinal permeability, attenuate nitrogen loss, improve survival, and lower the incidence of nosocomial infections. Previous studies in critically ill children were limited to very-low-birthweight infants and had equivocal results. METHODS: Eighty newborns and infants were included in a double-blind, randomized trial comparing standard parenteral nutrition (sPN; n = 39) to glutamine-supplemented parenteral nutrition (GlnPN; glutamine target intake, 0.4 g kg day; n = 41), starting on day 2 after major digestive-tract surgery. Primary endpoints were intestinal permeability, as assessed by the urinary excretion ratio of lactulose and rhamnose (weeks 1 through 4); nitrogen balance (days 4 through 6), and urinary 3-methylhistidine excretion (day 5). Secondary endpoints were mortality, length of stay in the ICU and the hospital, number of septic episodes, and usage of antibiotics and ICU resources. RESULTS: Glutamine intake plateaued at 90% of the target on day 4. No differences were found between patients assigned sPN and patients assigned GlnPN regarding any of the endpoints. Glutamine supplementation was not associated with adverse effects. CONCLUSIONS: In newborns and infants after major digestive-tract surgery, we did not identify beneficial effects of isonitrogenous, isocaloric glutamine supplementation of parenteral nutrition. Glutamine supplementation in these patients therefore is not warranted until further research proves otherwise. PMID- 15798462 TI - 2003 report of the intestine transplant registry: a new era has dawned. AB - SUMMARY BACKGROUND DATA: The intestine has been more difficult to transplant than other solid organs. We analyzed registry data to determine the scope and success of intestine transplantation in the current era. METHODS: All known intestinal transplant programs participated. Patient- and graft-survival estimates were obtained using the Kaplan-Meier product limit method and were analyzed with the Wilcoxon statistic. RESULTS: Sixty-one programs provided data on 989 grafts in 923 patients. Four patients were lost to follow-up. The short-gut syndrome was the most common primary indication for transplantation. Sixty-one percent of the recipients were < or =18 years. Proportionally more combined intestinal and liver transplants were performed in this group. More than 80% of all current survivors had stopped parenteral nutrition and resumed normal daily activities. A multivariate analysis of cases within the last 5 years revealed that transplantation of patients waiting at home, recipient age, antibody induction immune suppression, and center experience with at least 10 cases were associated with improved patient survival. One-year graft survival rates of 81% were achieved in patients who were induced with antithymocyte globulin and maintained on tacrolimus. CONCLUSIONS: Transplantation is an effective therapy for the treatment of patients with end-stage intestine failure who cannot tolerate parenteral nutrition. With newer immune suppressive protocols, 1-year graft and patient survival rates approach the results of liver transplantation. Further improvement in survival are expected with early referral since suitable donor organs are scarce and survival rates are better when patients are well enough to wait at home for their transplant. PMID- 15798463 TI - A total fundoplication is not an obstacle to esophageal emptying after heller myotomy for achalasia: results of a long-term follow up. AB - OBJECTIVE: The aim of this study was to evaluate the role and efficacy of a total 360 degrees wrap, Nissen-Rossetti fundoplication, after esophagogastromyotomy in the treatment of esophageal achalasia. SUMMARY BACKGROUND DATA: Surgery actually achieves the best results in the treatment of esophageal achalasia; the options vary from a short extramucosal esophagomyotomy to an extended esophagogastromyotomy with an associated partial fundoplication to restore the main antireflux barrier. A total 360 degrees fundoplication is generally regarded as an obstacle to esophageal emptying. MATERIALS AND METHODS: Since 1992 to November 2003, a total of 195 patients (91 males, 104 females), mean age 45.2 years (range, 12-79 years), underwent laparoscopic treatment of esophageal achalasia. Intervention consisted of Heller myotomy and Nissen-Rossetti fundoplication with intraoperative endoscopy and manometry. RESULTS: In 3 patients (1.5%), a conversion to laparotomy was necessary. Mean operative time was 75 +/- 15 minutes. No mortality was observed. Overall major morbidity rate was 2.1%. Mean postoperative hospital stay was 3.6 +/- 1.1 days (range, 1-12 days). At a mean clinical follow up of 83.2 +/- 7 months (range, 3-141 months) on 182 patients (93.3%), an excellent or good outcome was observed in 167 patients (91.8%) (dysphagia DeMeester score 0-1). No improvement of dysphagia was observed in 4 patients (2.2%). Gastroesophageal pathologic reflux was absent in all the patients. CONCLUSIONS: Laparoscopic Nissen-Rossetti fundoplication after Heller myotomy is a safe and effective treatment of esophageal achalasia with excellent results in terms of dysphagia resolution, providing total protection from the onset of gastroesophageal reflux. PMID- 15798464 TI - A multicenter evaluation of utility of chest computed tomography and bone scans in liver transplant candidates with stages I and II hepatoma. AB - OBJECTIVE: To determine utility of practice of chest computed tomography (CCT) and bone scan (BS) in patients with early-stage hepatoma evaluated for transplantation (LT). SUMMARY BACKGROUND DATA: Consensus-based policy mandates routine CCT and BS in LT candidates with hepatoma. No data exist either to support or refute this policy. METHODS: From January 1999 to December 2002, stages I and II hepatoma patients evaluated at 4 centers were included. Scan interpretation was positive, indeterminate, or negative. Outcomes of evaluation and transplantation were compared between groups based on scans. Total charges incurred were derived from mean of charges at the centers. RESULTS: One hundred seventeen stages I and II patients were evaluated. None had positive scans, 78 had negative, 29 had at least 1 indeterminate, and 10 did not have 1 or both scans. Twelve patients were declined listing, 6 from progression of hepatoma but none from CCT or BS findings. Two listed patients were delisted for progression of the hepatoma. Proportion of patients listed, transplanted, clinical and pathologic stage of hepatoma, and recurrence after LT were similar in groups with negative and indeterminate scans. Indeterminate scans led to 6 invasive procedures, 1 patient died of complications of a mediastinal biopsy, and none of the 6 showed metastases. Charges of $2933 were generated per patient evaluated. CONCLUSIONS: Positive yield of routine CCT and BS in patients with hepatoma is very low despite substantial charges and potential complications. CCT and BS performed only when clinically indicated will be a more cost-effective and safer approach. PMID- 15798465 TI - Current perceptions regarding surgical margin status after breast-conserving therapy: results of a survey. AB - OBJECTIVE: The surgical margin status after breast-conserving surgery is considered the strongest predictor for local failure. The purpose of this study is to survey how radiation oncologists in North America (NA) and Europe define negative or close surgical margins after lumpectomy and to determine the factors that govern the decision to recommend reexcision based on the margins status. METHODS: A questionnaire was sent to active members of the European Society of Therapeutic Radiation Oncology and the American Society for Therapeutic Radiology and Oncology who had completed training in radiation oncology. Respondents were asked whether they would characterize margins to be negative or close for a variety of scenarios. A second survey was sent to 500 randomly selected radiation oncologists in the United States to assess when a reexcision would be recommended based on surgical margins. RESULTS: A total of 702 responses were obtained from NA and 431 from Europe to the initial survey. An additional 130 responses were obtained from the United States to the second survey regarding reexcision recommendations. Nearly 46% of the North American respondents required only that there be "no tumor cells on the ink" to deem a margin negative (National Surgical Adjuvant Breast and Bowel Project definition). A total of 7.4% and 21.8% required no tumor cells seen at <1 mm and <2 mm, respectively. The corresponding numbers from European respondents were 27.6%, 11.2%, and 8.8%, respectively (P <0.001). Europeans more frequently required a larger distance (>5 mm) between tumor cells and the inked edges before considering a margin to be negative. CONCLUSION: This study revealed significant variation in the perception of negative and close margins among radiation oncologists in NA and Europe. Given these findings, a universal definition of negative margins and consistent recommendations for reexcision are needed. PMID- 15798466 TI - Prognostic importance of histologic vascular invasion in papillary thyroid carcinoma. AB - OBJECTIVE: To conduct a retrospective study of 39 patients with papillary carcinoma of the thyroid with histologic vascular invasion (VI+) and 361 patients without any sign of vascular invasion (VI-). SUMMARY BACKGROUND DATA: In the present study, we undertook a retrospective analysis of papillary carcinoma of the thyroid to assess whether histologically determined vascular invasion can be considered a predictive factor for prognosis. METHODS: By means of a retrospective study, we evaluated the department's database of patients with papillary thyroid carcinoma who had undergone total thyroidectomy from January 1993 to December 1999. RESULTS: Group I consisted of papillary carcinoma without any sign of vascular invasion (VI-) comprising 361 patients. Group II consisted of papillary carcinoma with vascular invasion (VI+) comprising 39 patients. At the time of diagnosis, we observed no metastases in patients with VI-, whereas a pulmonary metastasis was observed in 1 patient with VI+ (P = 0.0023). In 3.6% patients with VI- and in 20.5% patients with VI+, we observed recurrences in the regional lymph nodes (P < 0.001); we observed 6 (1.66%) distant metastases in patients with VI- and in the 12.8% patients with VI+ (P < 0.001). Three patients with VI+ (7.7%) and 2 patients with VI- (0.6%) died of tumor-related causes; these figures were found to be statistically significant (P < 0.001). CONCLUSIONS: In papillary carcinoma, it should be noted that histologic vascular invasion may be considered as a sign of an increased tendency toward hematogenic invasion and consequent increase in the relative percentage of metastases; ultimately, this means a poorer prognosis. In the presence of risk factors indicating a possible increase in biologic aggressiveness, adequate postoperative treatment and close follow up become essential. PMID- 15798467 TI - Gene expression changes with time in skeletal muscle of severely burned children. AB - OBJECTIVE: The purpose of this study was to identify gene-expression changes in leg muscle for up to 24 months after a severe thermal injury. SUMMARY BACKGROUND DATA: Hypermetabolism associated with severe burns was thought to cease with wound healing and closure. It has been recently shown that hypermetabolism does not completely resolve after healing, and muscle catabolism continues after hospital discharge; however, just how long after discharge has not been established. METHODS: : Six children, admitted to our hospital within 1 week after injury, were studied. Patients ranged in age from 3 to 18 years, with flame or scald burns covering more than 40% of their body surface area. At 1.5, 6, 12, 18, and 24 months postburn, a biopsy of the vastus lateralis muscle was taken and snap frozen at -80 degrees C. Total RNA was isolated and in vitro transcribed and hybridized to HG-U95 Av.2 Affymetrix arrays. The images were scanned and analyzed using Affymetrix GeneChip Analysis Suite 5.2 and dChip programs. Using 1 to 7 days after injury as baseline, comparisons were made of expression profiles at the various time intervals after injury. RESULTS: When comparisons are made to nonburned children, 38 genes were significantly altered at 1.5 months, 10 genes remained altered at 6 months, 4 remained altered at 12 months, and 2 at 18 months. No differences could be shown at 24 months. Western blot analysis of beta 2 microglobulin and myosin light chain was used to corroborate the microarray data. CONCLUSIONS: Gene changes can be identified for up to 18 months after burn but not at 24 months. These gene changes may provide information concerning what genes in skeletal muscle contribute to recovery from burn trauma. PMID- 15798468 TI - Risk factors for tissue and wound complications in gastrointestinal surgery. AB - BACKGROUND: Surgical site infections and disruption of sutured tissue are frequent complications following surgery. We aimed to assess risk factors predictive of tissue and wound complications in open gastrointestinal surgery. METHODS: Data from 4855 unselected patients undergoing open gastrointestinal surgery from 1995 through 1998 were recorded in a clinical database and validated. The database embraced variables related to patient history, preoperative clinical condition, operative findings and severity, and the surgeon's training. Variables predictive of surgical site infection and dehiscence of sutured tissue within 30 days after surgery were assessed by multiple logistic regression analysis. RESULTS: Following elective operation, the incidence of tissue and wound complications was 6% compared with 16% in emergency surgery (P < 0.001). These complications resulted in prolonged hospitalization in 50% of the patients and a 3-fold higher risk of reoperation but not increased mortality. Factors associated with complications following elective operations were smoking, comorbidity, and perioperative blood loss. Following emergency operations, male gender, peritonitis, and multiple operations were predictors of complications. Irrespective of elective or emergency surgery, the type of operation was a predictor of complications. CONCLUSION: Factors known to affect the process of tissue and wound healing are independently associated with tissue and wound complications following gastrointestinal surgery. PMID- 15798469 TI - Association of angiotensin-converting enzyme insertion/deletion polymorphism with serum level and development of pulmonary complications following esophagectomy. AB - BACKGROUND: Pulmonary complications remain the major cause of postoperative mortality in patients with esophageal cancer undergoing esophagectomy. It was unclear whether this dismal complication has a genetic predisposition. We therefore investigated the role of an angiotensin-converting enzyme (ACE) insertion/deletion polymorphism in developing these complications. METHODS: We conducted a prospective study including 152 patients with esophageal cancer who underwent esophagectomy in National Taiwan University Hospital between 1996 and 2002. The ACE genotype was determined by polymerase chain reaction amplification of leukocyte DNA obtained before surgery. The serum ACE concentration was determined by enzyme-linked immunosorbent assay. RESULTS: Thirty-five patients (23%) developed pulmonary complications following esophagectomy. As compared with patients with the I/I and I/D genotypes, those with the D/D genotype had a higher risk for pulmonary complications (adjusted odds ratio [OR], 3.12; 95% confidence interval [CI], 1.01-9.65). The risk was additively enhanced by combination of the ACE D/D genotype with other clinical risk factors (old age, hypoalbuminemia, and poor pulmonary function). The circulating ACE level was also dose-dependently with the presence of ACE D allele. As compared with the patients with circulating ACE less than 200 ng/mL, the patients with circulating ACE of 200 to 400 ng/mL and over 400 ng/mL had ORs (95% CI) of 2.75 (1.12-6.67) and 15.00 (4.3-52.34) to present with ACE D allele, respectively. CONCLUSIONS: An ACE insertion/deletion polymorphism might modulate the function of ACE gene and play a role in affecting individual susceptibility to pulmonary injury following esophagectomy in patients of esophageal cancer. PMID- 15798470 TI - The severity of injury in children resulting from acts against civilian populations. AB - OBJECTIVE: To characterize the injuries to children by acts against civilian populations (AACP). SUMMARY BACKGROUND DATA: Only 2 articles have focused on the spectrum and severity of injuries to children by AACP. METHODS: A retrospective case study of children 0 to 18 years old who were entered into the Israel National Trauma Registry as a result of AACP between September 29, 2000, and June 30, 2002. RESULTS: A total of 158 children were hospitalized for injuries caused by AACP, accounting for 1.4% of all hospitalized injured children but for 10.9% of all in-hospital deaths for trauma. Explosions injured 114 (72.2%); shootings, 34 (21.5%); and other mechanisms such as stoning or stabbing, 10 (6.3%). Older children were injured by explosions more frequently than younger children (86.1% of 15- to 18-year-olds, 73.7% of 10- to 14-year-olds, 63.2% of 0- to 9-year-olds, P = 0.02). A higher percentage of children injured by explosions rather than by shootings were severely or critically injured (33.9% versus 18.8%, P = 0.10). The most frequently injured body regions were extremities (62.8%), head/face (47.3%), chest and abdomen (37.2%), and brain (18.2%). More than 1 body region was injured in 63.0%. Children injured by explosions as compared with shootings had a lower incidence of abdominal trauma (14.9% versus 20.6%), a similar incidence of chest trauma (16.7% versus 14.7%), but a higher incidence of extremity trauma (65.8% versus 53.0%). There were 7 in-hospital deaths, 6 due to severe head injury and 1 due to severe abdominal trauma; 6 of the 7 deaths were caused by explosions, and all but 1 occurred in children 15 to 18 years old. CONCLUSIONS: AACP cause significant morbidity and mortality in children, especially adolescents. Injury severity is significantly higher among children who are injured by explosions rather than by shootings. PMID- 15798472 TI - In the forest of academia, are printed scientific journals an endangered species? PMID- 15798471 TI - Is primary resection and salvage transplantation for hepatocellular carcinoma a reasonable strategy? PMID- 15798473 TI - Cataracts and cataract surgery in mentally retarded adults. AB - PURPOSE: To examine the prevalence of cataracts and cataract surgery in a population of mentally retarded adults. METHODS: The prevalence of cataracts and cataract surgery was examined in a retrospective chart review. A total of 119 consecutive patients from the Southside Virginia Training Center for mentally retarded adults were seen in the authors' ophthalmology clinic during a 3-year period from January 1, 1999 to December 31, 2001. Intraocular lens (IOL) implantation and postoperative complications were noted in patients who had undergone previous cataract surgery. RESULTS: Of the 119 patients seen, 64 (54%) had cataracts or a history of cataract surgery. Thirteen (20%) of these 64 patients had a history of cataract surgery (21 eyes total). Seventeen (81%) of these 21 eyes had IOL implantation (14 posterior-chamber IOLs [PCIOLs] and three anterior-chamber IOLs). Postoperative surgical complications occurred in 7 (33%) of 21 eyes and included anterior uveitis in three (14%) eyes, posterior capsular opacification in two (10%) eyes, rhegmatogenous retinal detachment in one (5%) eye, dislocated PCIOL in one (5%) eye, pseudophakic bullous keratopathy in one (5%) eye, and cystoid macular edema in one (5%) eye. There were no cases of postoperative endophthalmitis or retained lens fragments. CONCLUSIONS: The prevalence of cataracts in adult mentally retarded patients is higher in this series (P<0.001) than in the world literature. However, if only visually significant cataracts are considered, the results are consistent with previous studies. Postoperative complications were common. Patients experienced higher rates of anterior uveitis (P=0.001) and pseudophakic bullous keratopathy (P=0.002) in this study than in previous studies. In this population, careful preoperative screening is recommended to select those patients most likely to benefit from cataract surgery and least likely to experience severe postoperative complications. PMID- 15798474 TI - Effects of quorum sensing molecules of Pseudomonas aeruginosa on organism growth, elastase B production, and primary adhesion to hydrogel contact lenses. AB - PURPOSE: Extracellular proteinase production and biofilm structure of Pseudomonas aeruginosa are affected by acylated homoserine lactones (AHLs) termed quorum sensing (QS) factors. C12- and C4-AHLs of P. aeruginosa and a synthetic furanone signaling antagonist were evaluated for effects on growth, primary adhesion to hydrogel contact lenses, and elastase B production of P. aeruginosa strains, including QS mutants, and isolates of Staphylococcus aureus and Staphylococcus epidermidis. METHODS: Acylated homoserine lactones and furanone were added to cultures of bacteria at various growth stages. Primary adhesion of P. aeruginosa to lenses soaked in AHLs and furanone was examined by adenosine triphosphate analysis. Elastase B activity of cultures exposed to AHLs and furanone was determined by an elastin congo red assay. RESULTS: Acylated homoserine lactones had no significant effects on maximal cell densities or primary adhesion. Adhesion of a lasI-rhlI- QS mutant to lenses was significantly lower than its wild-type (WT) strain, regardless of added AHL or furanone. Exogenous AHLs had negligible effects on elastase production of WT P. aeruginosa but restored elastase production in QS-deficient mutants. The furanone was biocidal for staphylococci but enhanced growth of the lasI-rhlI- QS mutant. The furanone decreased elastase production of WT strains but increased QS mutant elastase production. CONCLUSIONS: Exogenous AHLs and the furanone were of minor importance in initial adhesion of P. aeruginosa to lenses. The furanone decreased staphylococci growth and elastase production of WT P. aeruginosa but increased growth and elastase production of QS mutants. The data further indicate strain diversity in P. aeruginosa and complexity of AHL systems. PMID- 15798476 TI - Laser in situ keratomileusis treatment for myopia in a patient with partial limbal stem cell deficiency. AB - PURPOSE: The authors report a patient with soft contact lens-induced partial corneal limbal stem cell deficiency who successfully underwent laser in situ keratomileusis (LASIK) for correction of myopia. BACKGROUND: A 23-year-old man with a 6-year history of daily soft contact lens wear developed partial limbal stem cell deficiency of both corneas. The corneal disease resolved with cessation of contact lens wear and topical corticosteroid treatment. LASIK was performed in both eyes 1 year later. RESULTS: The patient successfully underwent LASIK and achieved an uncorrected visual acuity of 20/15 in both eyes. No complications were observed intraoperatively or postoperatively. The cornea and refraction remained stable 3 months postoperatively. CONCLUSION: In this patient with partial limbal stem cell deficiency secondary to soft contact lens wear, LASIK was a viable and safe alternative for the correction of myopia. PMID- 15798475 TI - Amoebicidal activity of multipurpose contact lens solutions. AB - PURPOSE: To compare the amoebicidal activity of two commercially available multipurpose contact lens care systems, by using cyst and trophozoite growth of two strains each of Acanthamoeba castellanii and Acanthamoeba polyphaga. METHODS: Trophozoites and cysts of Acanthamoeba species, isolated from corneal ulcers and an ATCC strain, were exposed to two commonly used multipurpose solutions and a saline control for 4 or 6 hours at ambient temperature. After neutralization of the disinfecting solution, track forming units were enumerated on Escherichia coli-seeded nutrient agar plates. RESULTS: There were significant differences (P<0.01) between solutions, strains, and cysts or trophozoites, but not between the different periods. Solution 2, containing polyquaternium-1 and myristamidopropyl dimethylamine (among other ingredients), gave greater mean log reductions than did solution 1, containing polyaminopropyl biguanide and poloxamine (among other ingredients). Both solutions generally reduced more trophozoites than cysts. CONCLUSIONS: The multipurpose solution containing polyquaternium-1 and myristamidopropyl dimethylamine reduced more cysts and trophozoites than did the alternative solution tested. Because there were differences in the effect of the multipurpose solutions on strains, it is recommended that more than one strain or species of Acanthamoeba be tested to determine efficacy of disinfecting solutions. The lack of difference between disinfection times may indicate that 4 hours could be considered as an adequate disinfecting time to ensure sufficient kill of Acanthamoeba species. PMID- 15798477 TI - The comparison of overnight lens modalities (COLM) study. AB - PURPOSE: The Comparison of Overnight Lens Modalities (COLM) Study is a controlled, randomized, clinical pilot study to determine the sample size required to perform a multicenter clinical trial comparing Paragon CRT lenses with CIBA Vision Focus NIGHT and DAY 30-day continuous-wear silicone hydrogel lenses. METHODS: Twenty subjects were enrolled in the study. Eighteen subjects, 8 CRT and 10 Focus NIGHT and DAY lens wearers, completed the 3-month study. Visual acuity and refractive quality of life were assessed. Nonparametric statistics were used to analyze differences within and between treatment groups for refraction, visual acuity, and refractive quality of life. The Wilcoxon rank sum test was used to analyze differences between the groups in refraction, visual acuity, and refractive quality of life. The Wilcoxon signed rank test was also used to monitor changes in refraction, refractive quality of life, and visual acuity within a group. Sample size calculations were performed to determine the sample size needed for a large-scale clinical trial. RESULTS: No statistically significant difference was observed between the groups for 12 of 13 scales on the National Eye Institute Refractive Error Quality of Life Instrument (NEI RQL-42) after treatment. A statistically significant difference was observed between the CRT and Focus NIGHT and DAY groups after 3 months for the NEI RQL-42 dependence on-correction scale (P=0.0032). There was a significant change within the CRT group between baseline and 3 months (P=0.0156). No statistically significant difference was observed between the two groups in Bailey-Lovie high- and low contrast visual acuity after 3 months. CONCLUSIONS: The NEI RQL-42 may be able to detect differences in refractive quality of life between two contact lens treatment groups for the dependence-on-correction scale. Paragon CRT and CIBA Vision Focus NIGHT and DAY lenses produced similar high- and low-contrast visual acuity in this study. A sample size of 126 subjects per group is required to find a 10-unit difference on the NEI RQL-42. PMID- 15798478 TI - Intraoperative correction of induced astigmatism after spherical correction of hyperopia with conductive keratoplasty. AB - PURPOSE: To evaluate the treatment of surgically induced astigmatism intraoperatively during conductive keratoplasty (CK) for correcting hyperopia. METHODS: Conductive keratoplasty uses radiofrequency energy applied to the peripheral corneal stroma to shrink the collagen and alter the central cornea to correct hyperopia. Nineteen consecutive patients (27 eyes) who underwent CK for hyperopia and were treated intraoperatively for induced astigmatism were examined. By using automated keratometric readings taken during the procedure, additional spots were applied at the minus cylinder or flat axis at the 7-mm zone until the intraoperative astigmatism was 2 diopters (D) or less. RESULTS: The intraoperative treatment reduced the astigmatism by an average of 2.30 +/- 1.32 D (P=0.00001). The mean induced astigmatism was 3.33 +/- 0.14 D for eyes that received eight spots, 4.12 +/- 1.13 D for eyes that received 16 spots, 4.43 +/- 0.82 D for eyes that received 24 spots, and 4.60 +/- 1.08 D for eyes that received 32 spots. Additional spots reduced astigmatism in most patients to less than 2 D. CONCLUSIONS: Intraoperative treatment of astigmatism through the addition of more spots at the minus cylinder or flat axis reduced the degree of induced astigmatism. Surgically induced astigmatism was observed more frequently in patients who received 32 treatment spots and 6-mm treatment zone application. PMID- 15798479 TI - The biomechanics of keratoconus and rigid contact lenses. AB - PURPOSE: To examine aspects of the genesis and progression of keratoconus through an analysis of the biomechanical forces associated with this condition, including those generated by rigid contact lenses. METHODS: The biomechanics of applanation tonometry and rigid contact lenses serve as a basis for examining contact lens adherence and the potential for contact lens induced mid peripheral corneal applanation and apical molding with apical clearance fittings. DISCUSSION: Some physiologic and pathologic mechanisms for increases in intraocular pressure are reviewed. The possibility is raised that hard squeeze blinks, for example, could significantly increase the distending forces that bear on the corneal apex in keratoconus and that some apical clearance contact lens fittings could increase the risk of progressive ectasia. Support for these hypotheses is found among healthy eyes and eyes with keratoconus that show increased curvature when apical clearance fittings are worn. CONCLUSIONS: The known risk of scarring responses to excessively flat fitting rigid contact lenses must be balanced against the possible risk of molding and ectasia advancement responses to tight definite apical clearance lenses. Fittings within the range of minimal apical clearance and minimal apical contact (divided support) may be the most appropriate. The appearance and performance of these fittings may be difficult to distinguish clinically. Because some patients may develop pathologic levels of intraocular pressure with vigorous eye rubbing, strong squeeze blinks, inverted body positions, and strenuous muscular effort, patients with, or at risk for, keratoconus, glaucoma, or progressive myopia should be advised of the possible adverse significance of these activities. PMID- 15798481 TI - Crystallization of gatifloxacin after penetrating keratoplasty. PMID- 15798484 TI - Nerves, reflexes, and the enteric nervous system: pathogenesis of the irritable bowel syndrome. AB - The bowel exhibits reflexes in the absence of CNS input. To do so, epithelial sensory transducers, such as enterochromaffin (EC) cells, activate the mucosal processes of intrinsic (IPANs) and extrinsic primary afferent (sensory) neurons. EC cells secrete serotonin (5-HT) in response to mucosal stimuli. Submucosal IPANs, which secrete acetylcholine and calcitonin gene-related peptide, initiate peristaltic and secretory reflexes and are activated via "5-HT1P" receptors. Release of neurotransmitters is enhanced by 5-HT4 receptors, which are presynaptic and strengthen neurotransmission in prokinetic pathways. 5-HT3 receptors mediate signaling to the CNS and thus ameliorate cancer chemotherapy associated nausea and the visceral hypersensitivity of diarrhea-predominant irritable bowel syndrome (IBS-D); however, because 5-HT3 receptors also mediate fast ENS neurotransmission and activate myenteric IPANs, they may be constipating. 5-HT4 agonists are prokinetic and relieve discomfort and constipation in IBS-C and chronic constipation. 5-HT4 agonists do not initiate peristaltic and secretory reflexes but strengthen pathways that are naturally activated. Serotonergic signaling in the mucosa and the ENS is terminated by a transmembrane 5-HT transporter, SERT. Mucosal SERT and tryptophan hydroxylase-1 expression are decreased in experimental inflammation, IBS-C, IBS-D, and ulcerative colitis. Potentiation of 5-HT due to the SERT decrease could account for the discomfort and diarrhea of IBS-D, while receptor desensitization may cause constipation. Similar symptoms are seen in transgenic mice that lack SERT. The loss of mucosal SERT may thus contribute to IBS pathogenesis. PMID- 15798485 TI - Visceral hypersensitivity. AB - Visceral hypersensitivity is considered one of the causes of functional gastrointestinal disorders. The objectives of this review are to provide a practical description of neuroanatomy and physiology of gut sensation, to describe the diverse tests of visceral sensation and the potential role of brain imaging to further our understanding of visceral sensitivity in health and disease. Changes in motor function in the gut may influence sensory levels, eg, during contractions or as a result of changes in viscus compliance. New insights on sensory end organs, such as intraganglionic laminar endings, and basic neurophysiologic studies showing afferent firing during changes in stretch rather than tension illustrate the importance of different types of stimuli, not just tension, to stimulate afferent sensation. These insights provide the basis for understanding visceral sensation in health and disease, which will be extensively discussed in subsequent articles. PMID- 15798486 TI - Functional chest pain: nociception and visceral hyperalgesia. AB - Functional chest pain is a common, yet poorly understood entity. The focus of this review is to explore the evolving research and clinical approaches with a particular emphasis on the sensory or afferent neuronal dysfunction of the esophagus as a key player in the manifestation of this pain syndrome. Although once regarded as a psychologic or esophageal motility disorder, recent advances have shown that many of these patients have visceral hyperalgesia. Whether visceral hypersensitivity is a central or peripheral perturbation of the gut brain axis remains debatable. Response to empirical therapy with high-dose proton pump inhibitors, upper endoscopy, or prolonged recording of esophageal pH may identify gastroesophageal reflux disease as a source of chest pain. Esophageal balloon distension study can serve as a useful test for identifying hypersensitivity. Newer techniques, including functional magnetic resonance imaging, magnetoencephalogram, and cortical evoked potentials, are being investigated. High doses of proton pump inhibitors and low doses of tricyclic antidepressants or trazadone remain the mainstay of therapy, although several new approaches including theophylline have been shown to be beneficial. PMID- 15798487 TI - Pathophysiology and treatment of functional dyspepsia. AB - Functional dyspepsia is a highly prevalent symptom complex and a heterogenous disorder. Recent studies showed potential associations between specific pathophysiologic disturbances and dyspeptic symptoms. Delayed gastric emptying reported in about 30% of patients with functional dyspepsia is associated with the symptoms of postprandial fullness, nausea, and vomiting. Impaired gastric accommodation present in 40% of functional dyspepsia patients is found to be associated with early satiety. Hypersensitivity to gastric distension is observed in 37% of functional dyspepsia patients and associated with the symptoms of postprandial pain, belching, and weight loss. Psychosocial factors and altered response to duodenal lipids or acid have also been identified as pathophysiologic mechanisms. Therapeutic options are still limited but targeted therapy directed at the underlying pathophysiology seems desirable. Thus, efforts to further elucidate underlying pathophysiologic mechanisms and identify the appropriate patient population using some type of pathophysiologic testing will be required. PMID- 15798488 TI - Functional biliary-type pain: update and controversies. AB - Recurrent biliary-type abdominal pain is a perplexing clinical dilemma that occurs in patients with an acalculous gallbladder in situ or in patients who have undergone a previous cholecystectomy. The pathogenesis of functional biliary-type pain is often unclear; therefore, evaluation and management remain controversial. In patients with an acalculous gallbladder in situ, critical importance has been given to delayed gallbladder emptying using cholescintigraphy (CCK-CS) to determine if gallbladder dysfunction is present. However, several issues remain unresolved, including methodology, definition of delayed emptying, and the absence of high-quality studies to determine if CCK-CS can predict who will do well with cholecystectomy. In patients with previous cholecystectomy, the main area of controversy is the evaluation of patients with sphincter of Oddi Type III, including the role of endoscopic retrograde cholangiopancreatography with SO manometry and sphincterotomy in these patients. Suggested algorithms for management of both clinical scenarios are provided. PMID- 15798490 TI - Irritable bowel syndrome: a syndrome in evolution. AB - As a group, functional gastrointestinal disorders are the most common gastrointestinal disorder seen by both generalists and specialists. These disorders can be frustrating to both patients and physicians as they are usually chronic in nature and difficult to treat. These disorders are associated with frequent healthcare visits, the scheduling of multiple, expensive diagnostic tests, and the use of both over-the-counter and prescription medications. All of these factors lead to a significant economic burden to society. In addition, functional gastrointestinal disorders are associated with a reduction in quality of life for the patient. Irritable bowel syndrome (IBS) is the most common of the functional gastrointestinal disorders. This syndrome has been the focus of a large number of research studies over the past two decades. These studies have resulted in a number of significant changes in our definition of IBS. In addition, these research studies have produced considerable changes in our understanding of the etiology and pathogenesis of IBS. In this section, we will review some of the evolutionary changes that have occurred in IBS. We will discuss how the definition of IBS has changed, consider our evolving strategies to evaluate and diagnose IBS, and finally, provide a brief overview of treatment options for this common disorder. PMID- 15798489 TI - Nausea, gastroparesis, and aerophagia. AB - Nausea, gastroparesis, and aerophagia are gastrointestinal phenomena that have variable impact on affected patients. The causes of nausea are varied; treatment of these conditions relates to the underlying etiology. Antiemetic agents acting on several distinct receptor subtypes produce benefits in distinct patient subsets. Gastroparesis is characterized by delays in gastric emptying, usually defined scintigraphically. Standard care of gastroparesis relies on dietary modification, antiemetic drug therapy, and initiation of medications that stimulate gastric motor activity. Recent advances include pyloric injection of botulinum toxin and surgical implantation of an electrical neurostimulator. Other surgical therapies are reserved for refractory cases. Aerophagia presents in individuals of normal and impaired cognitive function, most commonly with symptoms of overdistension or eructation. There are no pharmaceutical remedies for this condition; thus, therapy relies on behavioral treatments. PMID- 15798491 TI - Use of diet and probiotic therapy in the irritable bowel syndrome: analysis of the literature. AB - GOAL: The goal of this report is to review the use of dietary intake and probiotics in patients with irritable bowel syndrome (IBS) in published reports. BACKGROUND: Dietary factors can be important in inducing symptoms that occur in patients with the IBS. Dietary intolerances, dietary allergies, specific food metabolites, and regular diet contents all may act as triggers and aggravate the symptoms of IBS; but when any of these mechanisms can be proven to cause the symptoms, then their elimination results in the resolution of that patient's IBS. METHODS: Our previous review was updated. In addition, a careful Medline search was made for the years from 1975 to 2004 to evaluate human research reports on diet and probiotics in the IBS. Forty-six manuscripts were reviewed on diet and six were available on probiotic use in IBS. The most common dietary factor evaluated in the literature was bran, and the most common probiotic used was Lactobacillus plantarum. CONCLUSIONS: Although investigations have shown that bran may be helpful in some patients, a complete review of the literature does not reveal conclusive evidence that diet therapy is effective in IBS. From the limited reports on probiotics, there appears to be a trend to decreasing symptoms. It is clear that much more prospective research is needed to study both dietary factors and probiotics in these areas. PMID- 15798492 TI - Psychotherapeutics and serotonin agonists and antagonists. AB - Treatment of irritable bowel syndrome (IBS) remains challenging for patients and practitioners. Current therapeutic choices include antidepressants and psychotherapy, which are thought to target central nervous system triggers of symptoms. Data supporting these treatments are reviewed. Therapeutic agents targeted at receptors in the enteric nervous system have recently been developed to act locally in the gut. Alosetron, an antagonist for serotonin-3 receptors, reduces intestinal motility, secretion, and possibly sensitivity. It is effective for diarrhea predominant IBS, although there are some potentially serious side effects. Tegaserod, a serotonin-4 receptor agonist, is a prokinetic agent that speeds small bowel transit and right colon transit in IBS, reducing symptoms of constipation, pain, and bloating. IBS symptoms are improved with integration of old and new therapies, combined with reassurance, education, and lifestyle adjustments. PMID- 15798493 TI - What does the future hold for irritable bowel syndrome and the functional gastrointestinal disorders? AB - Our understanding of irritable bowel syndrome and the functional GI disorders has grown considerably over the last 15 years. In part this relates changes in their classification and definition from being due solely to motility disturbances, to being symptom based (eg, Rome criteria). This opened the door to the study of many other factors that contribute to the clinical expression of these disorders, including visceral hypersensitivity, sensitization, altered mucosal immunity, and dysfunction in brain-gut regulatory processes. New knowledge has been gained in areas of genetics, central nervous system and enteric nervous system neurotransmitters of motility, sensitivity and secretion, the effect of altered mucosal inflammation on cytokine and paracrine activation, and neural sensitization, postinfectious disorders, the influence of psychologic stress on gut functioning via alterations in regulatory pathways (eg, hypothalamic pituitary adrenal axis, or pain regulatory system like the cingulate cortex), improved accuracy of diagnosis using Rome II criteria plus "red flags" the institution of behavioral treatments, and the use of new pharmacologic treatments both at the gut and brain level. Future research will improve upon this new knowledge via basic and translational studies of neuropeptide signaling with new neurotransmitters, new knowledge on the mechanisms for central nervous system enteric nervous system communication and dysfunction, and more advanced clinical research on education, communication skills and their effects on outcome, genetics, pharmacogenetics and genetic epidemiology, better understanding as to how certain psychosocial domains (eg, catastrophizing, abuse) affect symptom behavior and outcome, newer pharmacologic treatments, and the use of combined pharmacologic and behavioral treatment packages. I am pleased to have the opportunity to provide a personal perspective on what the future will be for irritable bowel syndrome and the other functional GI disorders. Having been involved in this field for almost 30 years, I have been fortunate to witness tremendous changes. The focus of this presentation is to address the advances that have recently occurred that set the stage for proposing future research to help move the field along and ultimately to help our patients. PMID- 15798494 TI - Short stories. PMID- 15798495 TI - Diluent choice for subcutaneous infusion: a survey of the literature and Australian practice. AB - Continuous subcutaneous infusion is a method frequently used in palliative care to manage patient symptoms. To deliver the dose required and prevent subcutaneous sites from becoming inflamed and painful, the drug is often diluted in a solution, most commonly sterile water for injection or sodium chloride. The use of sterile water for injection has been recommended for cyclizine yet beyond this example there appears to be limited clinical direction regarding diluent selection. Inconsistency or lack of guidelines can be problematic if a diluent that may enhance the effectiveness of a drug compared with an alternate is not used because of lack of knowledge or guidance. This investigation considered existing literature, drug databases and directories, and involved a survey of palliative care services to examine evidence and experience relating to diluent selection. A number of inconsistencies emerged in both the literature and practice. With the exception of five drugs for which only saline was recommended, there appeared to be an inclination to use water unless contraindicated. Given an increasing reliance on this method of symptom management, the absence of formal clinical evidence or recommendations and ambiguity in relation to the use of diluents highlights the need for these deficits to be addressed as quickly as possible. PMID- 15798496 TI - Palliative care: roles and perceptions. PMID- 15798497 TI - Evaluation of a pilot nurse-led, community-based treatment programme for lymphoedema. AB - AIM: to evaluate the effect of this treatment programme on lymphoedema specific fluid volume, pain, discomfort, limb function and quality of life outcomes. DESIGN: prospective, descriptive study. SAMPLE AND SETTING: a convenience sample of four women with breast cancer related lymphoedema referred to a community based specialist oncology nurse consultant programme in Canada that involved manual lymph drainage combined with compression bandaging. MEASURES: circumferential arm measurements to assess fluid volume, visual analogue scales to assess the impact of lymphoedema on specific practical, physical and psychosocial problems known to be affected by lymphoedema and field notes. RESULTS: although a reduction in arm volume was observed, quality of life outcomes worsened for most of the women as they realized that this symptom would require lifelong management, serving as a reminder of their breast cancer experience. CONCLUSIONS: lymphoedema treatment programmes must ensure that the multidimensional impact of this symptom on women's lives is recognized and addressed as part of the treatment process. PMID- 15798498 TI - Pain and agitation in long-term care residents with dementia: use of the Pittsburgh Agitation Scale. AB - AIM: to explore the relationship between total and sub-scores of the Pittsburgh Agitation Scale (PAS) and five proxy measures of pain in long-term care (LTC) residents. STUDY DESIGN: descriptive correlational design. SAMPLE AND SETTING: 58 residents in three LTC facilities in rural Western Canada with moderate to severe cognitive impairment took part in the study. Six full-time registered nurses working in the facilities and three palliative care nurse consultants provided pain and agitation assessments. MEASUREMENTS: registered nurses used PAS to assess agitation. The five proxy measures of pain were the Discomfort Scale for Dementia of the Alzheimer's Type (DS-DAT), number of pain diagnoses, use of analgesic medications, and pain ratings by both facility nurses and palliative care nurse consultants. RESULTS: there was a moderately strong relationship between total PAS agitation scores and total DS-DAT pain scores (r=0.51, P<0.01). The PAS sub-score "resisting care" was significantly correlated with total DS-DAT scores (r=0.46, P<0.01), and pain ratings by both facility nurses (r=0.48, P<0.01) and palliative care nurse consultants (r=0.51, P<0.01). CONCLUSIONS: for certain residents with dementia, PAS may allow assessment of both agitation and uncommunicated pain. It is possible that the PAS form of agitation "resistance to care" may indicate pain that individual cannot otherwise communicate. One possible response to such resistance would be to trial pain medication and reassess agitation. Nursing staff in LTC facilities may need additional training in pain assessment of residents with dementia. PMID- 15798499 TI - Surprised endings: the dying of people with learning disabilities in residential services. PMID- 15798500 TI - Evaluation of training on the use of Graseby syringe drivers for rural nonspecialist nurses. AB - AIM: to assess the impact of a training programme on nurse confidence in: setting up the Graseby syringe driver (GSD); explaining the GSD to patient and family; setting the rate on the GSD; putting appropriate type and dose of drugs in the GSD. STUDY DESIGN: training programme with pre-training, post-training and follow up questionnaires. SAMPLE AND SETTING: palliative care nurse consultants presented half-day training sessions to 270 non-specialist nurses throughout the rural Grampians Health Region of Victoria, Australia. Nurses were from rural acute and sub-acute care settings, aged care facilities, and district nursing and nurse education services. MEASUREMENTS: demographic details of participants, previous experience and training with GSDs, comparative analyses of the four confidence parameters and participants' assessment of interest, new knowledge and usefulness of the training programme. RESULTS: increases in confidence levels were found in participating nurses in relation to each of the four confidence parameters. A follow-up survey tested residual benefit three months after the training programme. Statistically significant variations were found in nurses' confidence levels in relation to frequency of use. CONCLUSIONS: regular use of, and/or refresher sessions about the GSD are recommended to maintain optimum confidence, effective and safe nursing use of the GSD in palliative care. PMID- 15798501 TI - How basic is palliative care? A response to the responses so far. PMID- 15798502 TI - Report claims that a third of student nurses don't qualify. PMID- 15798503 TI - Nurses need advanced skills in disaster health care. PMID- 15798505 TI - The management of ballistic trauma: an infection control perspective. AB - This article discusses the relationship between ballistic trauma, clostridial contamination and potential wound infection and considers the implications for care by nurses and infection control teams. An overview of ballistics is presented followed by an examination of the pathophysiology of wounding and its effects. The philosophy of war surgery is balanced against civilian concepts and the differing management strategies that prevail. It explores the causes of, and relationship between, wound contamination and the seriousness of wound infection. Nurses, inexperienced in dealing and caring for these types of patients, should be aware and understand the beliefs, arguments and controversies that surround ballistic trauma management. By so doing, nurses and infection control teams will be able to provide an enhanced level of holistic nursing care. PMID- 15798506 TI - Exploring the role of an acute pain specialist nurse in the Royal Navy. AB - There is a strong research base to support the value and importance of acute pain management. There have been significant improvements in this specialty in the NHS since the introduction of acute pain teams in the early 1990s. The Armed Forces have embraced the concept more recently, and are making progress in this valuable area. This article examines the role of an acute pain specialist nurse in the Royal Navy and the importance of high quality, evidence-based pain management in the Armed Forces. The value of a military specialist nurse working as an integral part of a large NHS trust and how that relates to an operational role in the Royal Navy is also explored. The article draws conclusions about the importance of retaining senior specialist nurses in the clinical setting in order to ensure that the Armed Forces have access to up-to-date clinical expertise, especially when medical staff are deployed in operational roles. PMID- 15798507 TI - Personal experience of aeromedical evacuation of patients in the Balkans. AB - Nurses in the Princess Mary's Royal Air Force Nursing Service (PMRAFNS) undertake a variety of different roles in providing nursing care to Armed Forces personnel wherever they may serve. One such role is the evacuation by air of sick or injured personnel. This article discusses the experience of a nurse lecturer in the PMRANS who undertook the role of aeromedical evacuation liaison officer in the Balkans for a 4-month period during winter. The role is described and details of how the aeromedical evacuation of patients was organized is presented. The benefits of evacuating patients by air are discussed along with some of the potential problems that were encountered in the Balkans, including logistical problems and the vagaries of the weather. Finally, several examples of the types of patients evacuated are presented. PMID- 15798508 TI - Promoting quality sleep in older people: the nursing care role. AB - Sleep is a basic activity of living. Its restorative properties in healthcare settings--both hospital and community--should be considered by healthcare workers. Its significance in gerontological nursing should not be underestimated. As older people are prone to multiple pathology, disease processes may mask their need for effective sleep and rest. "Sleep architecture" is a term that applies to the mechanics of sleep or its stages and clinicians may fail to identify when this is disrupted. "Sleep hygiene" refers to measures or interventions used to promote sleep and a person-centred approach is necessary, with support from specialist agencies to achieve effective sleep hygiene. The need for long periods of sleep may diminish owing to ageing. However, the quality of the sleep experiences, i.e. feeling rested and refreshed, should be the same. Several factors have been identified in the causation of insomnia. In healthcare settings, environmental factors such as noise and light pollution are the commonest features. Other issues that may be present in both healthcare and community settings are related to varied perspectives, such as physiological, psychological and spiritual-religious. Providing a climate conducive to health sleep in all settings becomes a requisite in the planning of care. PMID- 15798509 TI - Property: procedures for nurses to follow after a patient has died. PMID- 15798510 TI - Rapid recovery following cardiac surgery: a nursing perspective. AB - Fast track or rapid-recovery pathways following cardiac surgery are becoming common practice in many cardiac units in order to maximize use of scarce critical care resources. Within the UK, rapid recovery generally describes same-day discharge from the initial intensive care facility to a lower-dependency unit. There are no nationally agreed protocols to help guide this practice. In a London teaching hospital a nurse-led audit was undertaken to identify which patients were selected for rapid recovery and to evaluate safety (length of hospital stay and incidences of postoperative complications) compared to a conventional recovery pathway. The study also sought to gain insight into the patients' views on rapid recovery. Data were collected on 104 patients, all patients (n = 56) who followed a rapid-recovery pathway were included. A comparison group (n = 48) was selected from patients who followed a conventional recovery but who were eligible for rapid recovery. The primary outcome, median length of hospital stay was 6 days for both groups, but the rapid-recovery group experienced significantly fewer postoperative complications. Rapid recovery as currently practised on this unit is safe for carefully selected cardiac surgical patients but barriers to rapid recovery need to be explored. PMID- 15798512 TI - Agency nurse who used the same speculum to carry out smear tests. PMID- 15798511 TI - The duty of nurses to meet patients' spiritual and/or religious needs. AB - As part of giving holistic care, nurses have a duty to meet the spiritual, religious and cultural needs of patients. These aspects of care are clearly identified in the Nursing and Midwifery Council's standards for nurses. According to the latest Census, 76.8% of people claim to have a religion. It is recognized that while people may not have a religious affiliation, they may have spiritual needs. The article considers the concepts of spirituality and religions and the significance of meeting these needs from the patient's perspective. Research based evidence demonstrates that patients' spiritual and/or religious needs are not always addressed by nurses. Barriers to giving spiritual/religious care are identified and spiritual assessment tools are considered. PMID- 15798513 TI - A balanced approach to dealing with violence and aggression at work. AB - The Government has introduced tough measures in dealing with violence and aggression in the workplace. Employers have a duty to provide adequate safety measures and training to reduce risks and ensure, where possible, the health, safety and welfare of their employees. This article highlights the responsibilities of employers and the pressures in both hospital and community environments on staff at risk from violent or abusive incidents. The patient's perspective is discussed, suggesting that more understanding between patients and staff is needed. The article looks at the "NHS Zero Tolerance Zone Campaign" and whether this can be enhanced by introducing training of an empathetic nature along with other practical environmental issues. It suggests that there are alternative and more effective, techniques in dealing with violence and aggression that can be used to defuse a situation before it ever becomes a physical altercation. The philosophies of eastern martial arts can teach us a lot about personal self-esteem and confidence which are two key elements in managing conflict situations. PMID- 15798514 TI - The importance of the ward team must be remembered. PMID- 15798515 TI - [The cellular and molecular basis of axonal growth]. AB - INTRODUCTION: During embryonic and post-natal development, numerous axonal connections are formed establishing a functional nervous system. Knowledge of the underlying molecular and cellular mechanisms controlling this phenomenon is improving. STATE OF THE ART: In this review, we present the general principles of axon guidance together with the major families of guidance signals. This includes the tyrosine kinase receptors Eph and their ligands Ephrins, the netrins, the semaphorins, the slits and other major components of the extracellular matrix. These types of guidance signals share common functional properties leading to actin cytoskeleton remodelling. The direct or indirect interactions between the receptors of these guidance cues and actin modulators is the final step of the signalling cascade constituting the fundamental mechanism defining the orientation and extension of the axonal growth cone. These factors are involved in the formation of many, if not all, axonal projections for which they act as repulsive (inhibitory) or attractive (promoting) signals. PERSPECTIVES: the knowledge of these mechanisms is particularly interesting since the inhibition of axonal outgrowth is considered to be one of the major obstacles to nerve regeneration in the central nervous system. Indeed, most of the guidance signals expressed during brain development are up-regulated in lesion sites where they contribute to the lack of nerve re-growth. Here, we present the nature of the mechanical barrier, the so called glial scar, and we describe the major inhibitory molecules preventing axonal extension. CONCLUSION: the comprehension of the molecular mechanisms involved in axon growth and guidance represents a major advance towards the definition of novel therapeutic strategies improving nerve regeneration. The path to the clinical application of these molecular factors remains long. Nevertheless, the next decade will undoubtedly provide challenging data that will modify the current therapeutic approaches. PMID- 15798516 TI - [Gliomatosis cerebri]. AB - INTRODUCTION: Gliomatosis cerebri (GC) is defined as a diffuse neoplastic glial cell infiltration of the brain involving more than two cerebral lobes and, occasionally, the infratentorial structures or the spinal cord. The tumor may appear de novo (primary GC) or result from the spreading of a focal glioma (secondary GC). Diagnosis and management of GC are difficult. Because of the diffuse nature of gliomatosis cerebri (GC), surgery is not suitable and large field radiotherapy carries the risk of severe toxicity. STATE OF ART: The analysis of current literature shows that the male population (58 percent) is younger, has a higher incidence of oligodendroglial GC and better prognosis than the female population. Survival (median=14.5 months) is also better for young patients, with high performance status, low-grade gliomatosis, and oligodendroglial subtype. Initial chemotherapy results in nearly 30 percent clinical or radiological improvement. In this setting, temozolomide is well tolerated and appears to be a valuable alternative to procarbazine-CCNU vincristine, especially for slow-growing, low-grade GC. PERSPECTIVE: Genotyping could be helpful to predict the response to chemotherapy in GC patients. PMID- 15798517 TI - [Megalencephalic leucoencephalopathy with subcortical cysts: a study of a Lebanese family and a review of the literature]. AB - INTRODUCTION: Megalencephalic leukoencephalopathy with subcortical cysts is a rare disease with autosomal recessive inheritance. MATERIALS AND METHODS: Two brothers born from a consanguineous marriage, presenting with the phenotype of the disease, their parents, brothers and sisters were examined. Magnetic resonance imaging of the brain was performed for the two patients. Sequence analysis of MLC1 (GenBank mRNA accession no. NM_OI5166) was performed for the patients using intronic primers. PCR restriction fragment length polymorphism analysis was done in patients, their parents and in 100 Lebanese controls in order to exclude gene polymorphism. RESULTS: The clinical features were characteristic of the disease, consisting of an early-onset macrocephaly followed by slowly progressive ataxia, pyramidal tract involvement and epileptic seizures. In one patient, the clinical manifestations were aggravated by a trivial brain trauma. In his brother and in one female cousin, a status epilepticus was precipitated by a febrile syndrome. The diffuse cerebral white matter lesions and the subcortical temporo-polar and frontal cysts, best seen on MRI, allowed making the diagnosis. Molecular genetics revealed a new mutation involving the MLC1 gene (263G-->T, exon 3). As a consequence, it affects the second transmembrane domain predict (G88V) of the MLC protein (protein sequence NP_055981). The mutation was confirmed by PCR restriction fragment length polymorphism analysis. CONCLUSION: Megalencephalic leucoencephalopathy with subcortical cysts may be individualized on clinical and radiological basis and confirmed by molecular genetics. In this Lebanese family, a new mutation of the MLC1 gene is reported. PMID- 15798518 TI - [Parakinesia brachialis oscitans]. AB - INTRODUCTION: In some cases of hemiplegia the onset of yawning is associated with an involuntary raising of the paralyzed arm. PATIENTS AND METHOD: Four observations of this movement, which is seldom described probably because it is mostly neglected, were made in the neurology unit of the University Hospital of Poitiers. The descriptions were compared with other cases that have been published in the medical literature of the last 150 years. Cerebral imagery shows a lesion that is most often localized on the internal capsule. After comparison with experimental models in cats, it is proposed that the section of the cortico neocerebellum tract of the extra-pyramidal system disinhibits the spino archeocerebellum tract, enabling a motor stimulation of the arm by the lateral reticular nucleus, which harmonises central respiratory and locomotor rhythms. RESULTS AND CONCLUSION: Some subcortical structures, that are phylogenetically more ancient, thus disinhibit regained autonomy in the homeostasis process associating the massive inspiration of yawning--a form of reflex behavior that stimulates vigilance--with a motor control that is active during locomotion. For this phenomenon we coined the term "parakinesia brachialis oscitans". PMID- 15798519 TI - [Decision making and executive function in severe traumatic brain injured patients: validation of a decision-making task and correlated features]. AB - OBJECTIVE: At the chronic stage, severe traumatic brain injured (TBI) patients experience difficulty in making decisions. Several studies have demonstrated the involvement of the prefrontal cortex, in particular the orbitofrontal region, in decision-making. The aim of the present study was to validate a decision-making task in this population and to ascertain whether the components of their dysexecutive syndrome may affect their decision-making and lead to difficulties for social rehabilitation. PATIENTS AND METHODS: Fifteen TBI patients and 15 controlled subjects matched for age, sex and years of education were assessed by a battery of executive tests (GREFEX) and by the gambling task (GT). RESULTS: The TBI subjects performed significantly worse than the controlled group in five out of six GREFEX tests. The TBI choices are significantly more disadvantageous than the choices of the control group when considering the three last blocks of 20 cards of the GT. The GT total score correlated significantly with execution time of the Stroop interference condition and the Trail Making Task B, as well as with the two measures (correct sequence span and number of crossed boxes) of the double condition of Baddeley's task. CONCLUSION: We postulate that executive functioning (supervisory attentional system) influence performance in the gambling task through mechanisms of inhibitory control, divided attention and working memory. Thus, this task seems to be determined by multiple factors; the process of decision-making may depend on frontal integrity. PMID- 15798520 TI - [Acetylcholinesterase inhibitor treatment for Alzheimer's disease: the experience in Picardy]. AB - INTRODUCTION: Despite numerous advances in the management of patients with Alzheimer's disease (AD), the proportion of patients treated according to current recommendations remains unknown. METHODS: In order to examine this point, we performed a study assessing the proportion of AD patients receiving one acetylcholinesterase inhibitor. This study was performed in Picardy (1.9 millions of inhabitants) in 2000 and 2001. The number of patients receiving one acetylcholinesterase inhibitor (tacrine, donepezil, rivastigmine and galantamine) was determined using data from health insurance and from pharmaceutical companies. RESULTS: The prevalence of AD was estimated to vary from 10751 patients in 2000, to 10990 in 2001. The number of treated patients was 1798 in 2000 and 2572 in 2001 and this corresponded to 16.7 percent (95CI: 2.5) and 23.4 percent (95CI: 3.4) of prevalent cases, respectively. Following the exclusion of patients with moderate to severe AD (estimated to represent 25 percent of patients), the proportion of treated patients reached 22.4 percent (95CI: 2.3) in 2000 and 31.3 percent (95CI: 3.1) in 2001. CONCLUSIONS: Despite a significant increase between 2000 and 2001, this study shows that less than a third of AD patients with mild to moderate dementia were treated with acetylcholinesterase inhibitors. Although numerous factors might have affected our estimations, this study shows that effective care of AD patients remains largely insufficient. PMID- 15798521 TI - [Rhinocerebral mucormycosis]. AB - INTRODUCTION: Mucormycosis is a rare and acute fungal infection which is frequently lethal. The rhinocerebral form is usually seen in diabetics, but other localizations may occur in severely immunocompromised subjects. OBSERVATION: We report the case of a sphenoidal sinusitis associated with a probable cavernous sinus thrombosis and carotid artery thrombosis with middle cerebral artery territory infarction. Diagnosis was made on histological examination following sphenoidotomy. Early medical and surgical treatment led to a good outcome. CONCLUSION: The diagnosis of rhinocerebral mucormycosis should be considered in the clinical setting of necrotic sinusitis and acute neurologic deficit in diabetic patients. Early diagnosis and treatment are crucial factors leading to a good outcome. PMID- 15798522 TI - [Behcet's disease associated with peripheral neuropathy]. AB - INTRODUCTION: The involvement of the peripheral nervous system in Behcet's disease is very rare. CASE REPORT: We report a case of a 47-year-old man with a six-year history of Behcet's disease and a two-year history of peripheral nervous system involvement. This patient presented with paraesthesia and weakness of the upper and lower limbs, diarrhea and erectile dysfunction. The electromyogram showed evidence of an axonal sensorimotor neuropathy and the nerve biopsy showed an axonal neuropathy. Routine blood tests were normal, there was no increase of serum creatine kinase, aspartate aminotransfease or lactate dehydrogenase and no signs of hyperthyroidism. Fibroscopy and colonoscopy showed no signs of entero Behcet. The patient was treated with prednisone, cyclophosphamide and carbamazepine with an improvement of paraesthesia. CONCLUSION: The mechanism of the peripheral neuropathy in Behcet's disease is still unknown, it might be due to vasculitis of the vasa nervorum or to the side effects of colchicine. Our report is particular by the association of sensorimotor and autonomic involvement of peripheral neuropathy in a patient with Behcet's disease. PMID- 15798523 TI - [Cyclosporin-induced toxic neuromyopathy]. AB - INTRODUCTION: Cyclosporine is an immunosuppressive treatment whose side effects limit its usefulness. Among neurological side effects, neuropathies or myopathies have been reported, specially inpatients given combinations of cyclosporine with co-enzyme A reductase inhibitors. CASE REPORT: We report here the case of a 67 year-old woman who developed few months after a kidney graft sensorimotor disorders which progressed rapidly. Since all etiologies of such a disorder were ruled out, the hypothesis of toxicity exclusively induced by cyclosporine was suggested and confirmed by the improvement observed after its withdrawal. CONCLUSION: This observation highlights the fact that cyclosporine may induce neuromyopathies even when given alone at the therapeutic dosage. PMID- 15798524 TI - [Relapsing ophthalmoplegia as a presenting symptom of orbital myositis]. AB - INTRODUCTION: We report a case of orbital myositis revealed by a relapsing and initially painless ophthalmoplegia. CASE REPORT: A 50-year-old patient suddenly presented with an isolated palsy of the right lateral rectus muscle which recovered in 3 weeks after a short-lasting corticosteroid therapy but relapsed. Cerebral and orbital imaging were normal. Orbital pain with a fluctuating course occurred 6 months later. Two years after the clinical onset, orbital CT scan revealed a fusiform hypertrophy of the right lateral rectus muscle which was enhanced by contrast. After corticosteroid treatment, the symptoms improved and CT scan became normal. CONCLUSION: Orbital myositis may not be ruled out in case of painless ophthalmoplegia and systematic and repeated imaging is necessary. PMID- 15798525 TI - [Relapsing remitting multiple sclerosis or multiphasic disseminated encephalomyelitis?]. AB - INTRODUCTION: The imaging presentation of some forms of multiple sclerosis may be misleading. In patients with a history of recent infection or vaccination, especially for adolescents or young adults, the differential diagnosis with acute disseminated encephalomyelitis can be difficult. CASE REPORT: We report an unusual clinical and radiological presentation of multiple sclerosis, mimicking acute disseminated encephalomyelitis. We discuss clinical and radiological differential diagnosis, and the outcome after immunosuppressive treatment. CONCLUSION: Distinguishing between acute disseminated encephalomyelitis and the first relapse of multiple sclerosis can be difficult. Brain imaging is a precious tool for differentiating between the two diseases. PMID- 15798526 TI - [The role of MRI in the diagnosis of progressive supranuclear palsy]. PMID- 15798527 TI - [Statins and prevention of strokes]. AB - Statins have become a leading prescription drug in France. Indications have been greatly extended over the last five years subsequent to publication of many multicentric prospective trials. Besides their preventive properties for coronary artery disease, their efficacy for the prevention of stroke also appears to be demonstrated in coronary patients (CARE and LIPID studies) and in subjects at risk (HPS and ASCOT-LLA studies) irrespective of the serum cholesterol level, the relative risk of stroke being decreased by 30 percent compared with placebo. This preventive capacity would involve pathways other than cholesterol and/or LDL cholesterol lowering mechanisms, and would probably involve a direct effect on inflammation, coagulation, modulation of certain atherogenic processes, and endothelial function. There remains however certain questions concerning the exact role of statins on secondary prevention of stroke (SPARCL study), their real usefulness in the elderly subject (PROSPER study), and the relative superiority of individual statins. PMID- 15798528 TI - [Should systematic prophylaxis following exposure to sex be proposed in French Guiana?]. PMID- 15798529 TI - [Determination of early markers of a sanitary event. The example of the heat wave of August 2003 at the Samu 93-centre 15 in France]. AB - OBJECTIVE: The aim of this study was to determine, a posteriori, the parameters detecting an event in a French medical emergency dispatching centre (SAMU). METHODS: Six parameters were retained: total number of medical requests received by the Samu 93-centre 15: the number of decisions to send a mobile intensive care unit (MICU), number of decisions to send a non-medical unit, number of decisions to send a general practitioner and number of deaths observed by the physicians of the MICU. For each parameter, a daily referential was established over the five previous years (1998 to 2002) and compared with the results of August 2003 RESULTS: The number of decisions to send a non-medical unit and the number of decisions to send a general practitioner were unchanged. The number of deaths on the 8th of August observed by the MICU physician should have led to an alert being given: 5.0 deaths for a referential of 1.7 (+ 194%). The number of decisions to send an MICU on the 7th of August should have led to an alert being given: 41 interventions for a referential of 25 (+64%). The number of medical interventions on the 6th of August should have led to an alert being given: 351 interventions for a referential of 299 (+17%). CONCLUSION: The total number of medical interventions treated by the SAMU 93-centre 15 is a the most sensitive and earliest marker of a sanitary event, such as that observed in August 2003 with the heat wave. PMID- 15798530 TI - [The editorial quality of hospital drug prescriptions in the Ile de France area. A study by the Health Insurance]. AB - OBJECTIVES: To assess the editorial quality of drug prescriptions made by hospital for ambulatory patient (outpatients or on discharge from hospital). METHODS: This study was based on 500 drugs prescriptions with bills which were presented to the National Health Scheme for reimbursement during July 2000, from hospital centres within the Ile de France area. In a second phase, the sub-group of prescriptions for patients on discharge from hospital were analysed and permitted comparison of the results obtained with those of a study conducted in a university hospital centre in 1997 on this type of prescription. RESULTS: In most cases, these prescriptions were drawn up during a medical consultation at the hospital (57%), on discharge from hospital (25%), or after a visit to a department of emergency unit (10%). The principal mistakes in editorial modalities were the omission of compulsory identification elements for the patient, the practitioner and the health care facility: only 17% of the prescriptions associated all these elements. In 41% of the cases, the identification of the hospital was missing, the prescriber's name in 18% and his/her function in 64%, the patient's family name was missing in 6% of the cases, and first name in 31%. Analysis of the contents of the drug prescriptions showed that 95% bore sufficient detail for the drug to be dispensed. The non respect of rules regarding the duration of prescriptions for anxiolitics (23%) and notably hypnotics (62%) was observed. Potentially dangerous drug interactions and contraindications concerned 8% of the prescriptions studied. Comparison with other studies on the subject in the Ile de France area showed converging results regarding errors made and the absence of any improvement in editorial quality since 1997. CONCLUSION: This study underlines the poor editorial quality of hospital prescriptions. The development of a standardised prescription and promotion of a computerised system would reduce the number of errors that may result from such prescriptions. PMID- 15798531 TI - [In situ mammary duct carcinoma with microinvasion. Which axillary lymph node exploration?]. AB - OBJECTIVE: Specify the role of axillary lymph node removal in micro-invasive in situ duct carcinomas (DCIS-MI) of the breast with a series of 107 consecutive cases. METHODS: Single-centre, retrospective, anatomoclinical study with application of the European guidelines adopting as pathological definition the presence of areas of micro-invasion not exceeding 1mm. Axillary lymph node dissection was systematically complete and was preceded by the search for the sentinel node in 10 patients using the isotope method. RESULTS: Lymph node invasion was revealed in 8 cases in the global population (7.5%). All the cases except one exhibited typical deleterious histological features: comedo architectural sub-type, high nuclear grade, and size of the lesion>3 cm. The lymph node invasion was of 18.5% in the sub-group of high-grade micro-invasive comedo-carcinomas measuring more than 3 cm. CONCLUSION: Contrary to those exhibiting pure DCIS, DCIS-MI patients require surgical exploration of the armpit, the most appropriate modalities of which are currently debated: classical axillary lymph node dissection or search for the sentinel lymph node; the selective lymphadenectomy procedure is not yet a consensually validated technique. PMID- 15798532 TI - [Retroperitoneal fibrosis]. AB - OBJECTIVE: We studied the clinical, therapeutic and progressive characteristics of retroperitoneal fibrosis. METHODS: We analysed the observations of retroperitoneal fibrosis diagnosed between 1980 and 2002 in our hospital, from the summaries of 15 patients exhibiting retroperitoneal fibrosis (RPF). Therapeutical supervision was based on biology and radiology. RESULTS: There were 11 men and 4 women with a mean age of 44.5 years ranging from 28-64 years. Pain was predominantly lumbar or abdominal in all patients. An inflammatory syndrome was observed in all patients and renal failure in 11. Radiological examinations revealed single or bilateral in 14 cases and the fibrosis plaque in 13 cases. Treatment consisted in corticosteroids alone in 9 patients, surgery alone in 3 cases and surgery with corticosteroids in 3 patients. Ten relapses (range: 1-5) occurred in 4 patients when corticosteroids were stopped. After a mean follow-up of 36 months (range:18 days-11 years), one death was observed, 12 patients had normal renal function and 2 patients had persistent moderate renal failure. CONCLUSION: This study confirms the rarity of retroperitoneal fibrosis, the difficulty in its diagnosis, the frequency of pain, inflammatory syndrome and renal failure. Corticosteroids are efficient and regular follow-up is required. PMID- 15798533 TI - [Mucormycosis in the diabetic patient]. AB - INTRODUCTION: Mucormycosis is a rare, devastating, fungal infection, which disproportionately affects non-controlled diabetic patients, notably during ketoacidosis. It can be manifested in rhinocerebral, but also pulmonary and disseminated forms. OBSERVATIONS: Four consecutive diabetic patients who were admitted to the Farhat Hached Hospital, Sousse, Tunisia, between January 2001 and November 2002, are presented. Three patients exhibited ketoacidosis and one renal failure with hyperosmolarity. Infection was limited to the sinuses in two cases, to lower respiratory tract in one case, and was probably disseminated in one case. Diagnosis was confirmed by mycological and histological findings in all cases. Systemic Amphotericin B was associated with surgical debridement of the lesions in patients with rhinocerebral involvement. DISCUSSION: Despite aggressive therapy, mortality was high (3 out of 4 patients). Mucormycosis remains a severe, frequently fatal disease in diabetic patients. PMID- 15798535 TI - [Hodgkin's disease with bone marrow involvement. The role of positron tomography]. AB - INTRODUCTION: Hodgkin's disease with initially predominant bone marrow involvement is observed in 1% of cases. OBSERVATION: A case of Hodgkin's disease with massive bone marrow invasion, manifested by pancytopenia in the context of fever and alteration in general status of health, with little or non superficial peripheral lymph nodes. The bone marrow biopsy revealed extensive myelofibrosis with rare Reed-Sternberg cells. DISCUSSION: This case report is inscribed within the framework of the exceptional massive bone marrow forms of Hodgkin's disease, described by Duhamel et al. in 1979. We believe that metabolic imaging with 18F FDG positron emission tomography (PET) is a valid examination in orienting diagnosis and post-therapy assessment. PMID- 15798534 TI - [Pancreatic peritoneal fistula with bisalbuminemia]. AB - INTRODUCTION: Pancreatic fistulas are a complication that occur in 3 to 15% of cases during the progression of chronic or acute pancreatitis, usually alcohol induced. Bisalbuminemia is characterised by two albumin fractions on serum protein electrophoresis. The presence of Bisalbuminemia is inconsistent and has only rarely been reported. OBSERVATION: A 42 year-old man, excessive drinker, developed pancreatic ascites related to a pancreatic-peritoneal fistula and associated with transitory bisalbuminemia. Treatment was medical with good short term results. DISCUSSION: Ascites was secondary to a pancreatic-peritoneal fistula. It can be constitutional or acquired and transitory, and secondary to prolonged treatment with b-lactamines in a patient with kidney failure or a pancreatic fistula. PMID- 15798538 TI - [Physiopathology of pulmonary arterial hypertension. Cellular and molecular aspects]. AB - The combined effects of vasoconstriction, remodelling of the pulmonary vessel walls and in situ thrombosis contribute to the increase in pulmonary vascular resistance during pulmonary arterial hypertension. Vascular remodelling involves all the sheaths of the vessel wall and all the cell types of which it is composed (endothelial cells, smooth muscle cells, fibroblasts, inflammatory cells and platelets). Excessive vasoconstriction has been related to a defect in the function of expression of the potassium channels and endothelial dysfunction. This leads to chronic insufficiency in the production of vasodilators, notably nitrogen monoxide and prostacyclin and the excessive production of vasoconstrictors such as endotheline-1. These defects contribute to the increase in vascular tonus and pulmonary vascular remodelling and represent pertinent pharmacological targets. Certain growth factors, including those of the super family of transforming growth factor beta, angiopoietine-1 and serotonin, may play a part in the pathogenesis of pulmonary arterial hypertension. PMID- 15798539 TI - [Lupus panniculitis]. AB - Lupus panniculitis is characterized by a T-cell lymphocyte infiltration of the fat, with fibrosing and cicatricial progression. The lesions are predominantly found on the head and the upper part of the body; they are more frequent in young women. Diagnosis is based on the integration of the clinical and histological data, which renders a deep cutaneous biopsy indispensable. Histopathology is the key to the differential diagnosis, which essentially includes subcutaneous T-cell lymphoma. The lupus panniculitis lesions can be isolated or, more often, integrated within a known lupus disease, usually of good prognosis. Treatment relies above all on synthetic anti-malarials, occasionally associated with local or general corticosteroids. PMID- 15798537 TI - [Guidelines for good practices should be based on clinical proof]. PMID- 15798540 TI - [Role of LDL in cardiovascular diseases. New data and new recommendations]. AB - The management of hypercholesterolemia has rapidly progressed with the large clinical trials mainly on statins. Over the past 15 years, it has been demonstrated that the principle target was LDL-cholesterol and that treatment with statins decreased cardiovascular morbidity and global mortality. The studies have progressively targeted patients with increasingly lower cholesterol levels but always at high risk. Recent data are in favour of the so-called "aggressive" strategy aimed at reducing cholesterol to very low levels. The emergence of new therapies with stronger statins and the concept of combining lipid-lowering treatments has improved the chances of reaching this aim. Prompted by this progress, the transposition of this aggressive strategy to clinical practice is confronted with many problems, which stem from practitioners' and patients' reticence and fears (raising the question of compliance to treatment) and, more generally, the problem of cost. In this perspective, therapeutic education is one of the tools that would promote compliance. New treatments in the field of HDL would further support the progress made in such prevention. PMID- 15798541 TI - [Foreign bodies in the soft tissues. The interest of sonography]. PMID- 15798543 TI - [The control of asthma: a vital sign to be assessed on each consultation]. PMID- 15798544 TI - [Medical and socio-economical impact of drug-induced adverse reactions]. AB - OBJECTIVE: To analyse the nature of adverse drug reactions, their frequency, the drugs incriminated, their outcome, the different ways to treat them and their cost. METHODS: Descriptive, prospective and non-comparative study, based on spontaneous reporting. Each patient was followed-up over a 6-month period. RESULTS: One hundred and fifteen adverse events were reported in 113 patients (58 men and 55 women), with a mean age of 59.5 years. Cutaneous and haematological reactions predominated, with a majority of cardiovascular and psychotropic agents incriminated. Sixty-three percent of adverse reactions were serious: 49% led to (or prolonged) hospitalisation, 6% led to death, 4% to after effects, 3% were life-threatening and 1% led to congenital malformation. The mean duration of an adverse reaction was of 37.4 days. The total cost of these 115 adverse reactions was an estimated 610.110 Euros, with a mean global cost of 5.305 Euros per event. CONCLUSION: This study shows the individual and collective extent of the impact of adverse individual and collective reactions. PMID- 15798545 TI - [Impact of the consensus conference on the ambulatory treatment of bronchiolitis in infants]. AB - OBJECTIVE: To assess the impact of guidelines of the consensus conference in September 2000 on the ambulatory management of acute bronchiolitis in infants. METHODS: Prospective multicenter study, in four hospitals in the North department in France, during three epidemic periods: the winter preceding the consensus conference and the two following winters. All the infants between 30 days and 2 years of age, admitted to an emergency care unit for a first or second episode of bronchiolitis were included. The infants' characteristics and ambulatory treatments prescribed were collected. RESULTS: Six hundred thirty-eight infants were included: 169 before the conference and 469 after. Seventy-seven percent had consulted previously and a prescription had been drawn-up for 77%. There was no difference in the frequency of drug prescriptions or physiotherapy between the three periods. After the conferences, inhaled beta(2) agonists were still prescribed in 21% of cases, inhaled corticosteroids in 15%, oral corticosteroids in 34%, antibiotics in 53% and mucolytics in 54%. Physiotherapy was prescribed for 58% of the infants. CONCLUSION: Two years after the consensus conference, the ambulatory treatment of bronchiolitis has no changed, corticosteroids, beta(2) agonists, antibiotics and mucolytics are still prescribed in excess. Further studies, within a few years, are required to reassess the application of the guidelines, not only in outpatient but also in inpatient management. PMID- 15798547 TI - [Acquired childhood and juvenile subclinical hypothyroidism with or without goitre]. AB - OBJECTIVE: Primary acquired childhood and juvenile hypothyroidism is mainly due to chronic thyroiditis or to the after effects of cervical radiotherapy for cancer. In the presence of clinically evocative signs, with a palpable goitre or firm thyroid, hypothyroidism is searched for. However, cases of childhood and juvenile hypothyroidism without goitre have been described. METHOD: We retrospectively studied childhood and juvenile sub-clinical hypothyroidism in those in whom antithyroid antibodies had been searched for and thyroid ultrasonographic examination had been performed. RESULTS: In this age group, we studied 34 cases of subclinical hypothyroidism (plasma TSH: 3.6-20 microIU/ml, normal free thyroxin level). In a first group (17 girls/1 boy, mean age: 14.4 years), 50% of patients had an abnormal palpable thyroid and 100% have auto immunity markers. In the other group (7 girls/9 boys, mean age: 12.0 years), there was no goitre and no auto-immunity marker had been found. The clinical signs that prompted consultation were identical in the two groups and, if no goitre existed, measurement of TSH levels established the diagnosis of the onset of hypothyroidism. DISCUSSION: It seems that second group patients constituted an unusual entity: predominance of boys, thyroid usually small and lack of auto immunity markers. PMID- 15798546 TI - [Compared effects of etomidate and propofol for anaesthesia during electroconvulsive therapy]. AB - OBJECTIVE: Electroconvulsive therapy (ECT) requires repeated short-term anaesthesia with muscle relaxation and deep narcosis and uses several anaesthetic agents. The aim of this study was to assess the quality of the anaesthetic technique applied for ECT by comparing two products: propofol and etomidate. METHODS: This was a prospective randomised study that included ECT sessions. Patients were distributed into two groups. Patients of Group 1 underwent general anaesthesia with propofol (1.5mg.kg(-1)) and succinylcholine (0.75 mg.kg(-1)). Patients of Group 2 were administered etomidate (0.15 mg.kg(-1)) and succinylcholine (0.75 mg.kg(-1)). None of the patients included had any absolute or relative contraindication to ECT. RESULTS: 104 sessions were included, with 52 sessions per group. Group 1 was composed of 12 patients and Group 2 of 13. The demographical characteristics and indication for ECT were comparable in the two groups. There was no haemodynamic variation (notably drop in blood pressure) between the groups. The duration of seizures was significantly more prolonged in the etomidate group (28.76 +/- 3.29 seconds) than in the propofol group (23.84 +/ 7.18 seconds), with significant difference (p = 0.000018). Awakening was calm in both groups. CONCLUSION: The pharmacological properties of propofol and etomidate reply precisely to the requirements of anaesthesia for ECT. Nevertheless, no drop in blood pressure was observed with the greater prolongation of seizures in the etomidate group compared with the propofol group. PMID- 15798548 TI - [Cogan's syndrome revealed by haemorrhagic glairous diarrhoea]. AB - INTRODUCTION: Cogan's syndrome is defined by the combination of non syphilitic interstitial keratitis and inner ear dysfunction, similar to Meniere's disease. OBSERVATION: Cogan's syndrome was revealed by haemorrhagic glaireous diarrhoea in a 21 year-old woman whose medical history included Hirschsprung's disease. Cerebral MRI revealed viral-like bilateral neuritis of the eighth cranial nerve, never described in the literature before. The auditory and visual disorders regressed after treatment with corticosteroids. CONCLUSION: The early diagnosis of Cogan's syndrome permits the cure of the visual and auditory and notably inner ear symptoms with corticosteroid therapy. PMID- 15798549 TI - [Myocardial localisation of tuberculosis: the diagnostic value of cardiac MRI]. AB - INTRODUCTION: Tuberculosis can be responsible for myocardial damage, the frequency of which is probably underestimated because of the difficulty in its diagnosis. We studied the contribution of cardiac magnetic resonance imaging (MRI) in three patients. OBSERVATIONS: Three patients were treated for disseminated tuberculosis. They had moderate cardiac abnormalities (tachycardia, dyspnoea on effort). The electrocardiogram was normal in 2 patients and the echocardiography showed localized hyperkinesias. Cardiac MRI revealed intra myocardial nodular gadolinium enhancement and hyperkinesias. The clinical outcome in the 3 patients was favourable following anti-tuberculosis therapy; one patient was also administered corticosteroids. DISCUSSION: Cardiac MRI is a non-invasive examination that brought important arguments for the diagnosis of tubercular myocarditis in the 3 patients. PMID- 15798550 TI - [Bartonella hensalae encephalopathy]. AB - INTRODUCTION: Bartonella hensalae is a poorly known cause of encephalopathy in young subjects. OBSERVATION: A 17 year-old adolescent was admitted in a state of emergency because of frequent convulsive seizures and inter-critical drowsiness. The diagnosis of encephalopathy was made on the association of these clinical signs and electro-encephalographic abnormalities. The presence of a cat in his home, a right axillary lymph node that had appeared in a context of fever, and positive serological kinetics related this encephalopathy to a bartonellosis. The course was good. DISCUSSION: Diagnosis of a Bartonella hensalae encephalopathy is based on a range of anamnesic, clinical and microbiological arguments. The potential interest of antibiotic therapy and its modalities remains to be established. PMID- 15798551 TI - [Oesophageal ulcer associated with the use of bacampicillin]. AB - INTRODUCTION: Many drugs may provoke oesophageal disorders. In France, doxycyclin, potassium chloride non-steroidal anti-inflammatories are the most frequent agents incriminated. OBSERVATION: A 27 year-old man consulted for dysphagia and odynophagia due to oesophagitis following administration of bacampicillin. COMMENTS: Several drugs can provoke oesophageal disorders including ulcerations. Predisposing circumstances exist. Ingestion of the drugs with a sufficient quantity of water (60 to 100 ml), whilst standing or sitting should be able to limit this type of event. PMID- 15798552 TI - [The 308 nm excimer laser in dermatology]. AB - THE EFFICACY OF THE 308 NM EXCIMER LASER in the treatment of common psoriasis has been demonstrated. THE DOSES USED have progressively decreased, hence, limiting the adverse events that appear redhibitory with high doses. THE ADAPTATION OF THE DOSES not to the patients themselves but to each of the plaques treated should reduce the number of sessions and the cumulated close necessary to obtain clinical remission. THE 308 NM EXCIMER LASER is effective and tolerance is good in the treatment of vitiligo. It should be proposed for limited vitiligo and essentially of the "UV sensitive" areas, which have shown aesthetically correct percentage of repigmentation. THE PLACE AND INTEREST of its association with other treatments, notably with topical tacrolimus, remains to be defined. Although the results obtained in the treatment of vitiligo are promising, they have to be confirmed in larger cohorts and ensure the absence of median and long term side effects. This therefore limits its use in combined treatments in the context of controlled clinical traits. THE 30 NM EXCIMER LASER IS AN EFFECTIVE AND WELL TOLERATED TREATMENT in localised and non-nodular forms of mycosis fungoid (MF). Although the number of patients treated is limited, the clinical and histological cure observed demonstrates the interest of this new technique in the treatment of MF. These results must be confirmed in a greater number of patients. THE 308 NM EXCIMER LASER is an interesting therapeutic alternative in the treatment of plaques of alopecia areata, erosive oral lichen planus, post surgical hypopigmentation, vergetures and localised forms of atopic dermatitis. Because of the sparcity of data and in the absence of long term follow-up, it must not be proposed in first intention. PMID- 15798553 TI - [Therapeutic intensification and autologous stem cell transplantation in autoimmune diseases]. AB - THE PATHOPHYSIOLOGY of most autoimmune diseases is often poorly understood. EXPERIMENTAL CONSIDERATIONS and clinical experience suggest that high doses immunoablation followed by stem cell transplantation is a therapeutic option to consider for certain severe autoimmune disorders. THE CONCEPT OF RESTORING NORMAL IMMUNE REACTIVITY must in part br true since current results of 466 transplants (445 autologous, 21 allogeneic) patients suffering from various autoimmune diseases show a beneficial outcome in approximately 2/3 of the patients. TO IMPROVE THE EFFICACY AND SAFETY OF SUCH AN AGGRESSIVE PROCEDURE in patients with potentially affected vital organs by the underlying autoimmune disease, it is especially important to follow international consensus guidelines and to centrally collect clinical data for in depth analysis in the EBMT International Stem Cell Project for Autoimmune Disease in Basel, Switzerland. PHASE III STUDIES ARE RUNNING FOR SYSTEMIC SCLEROSIS (Astis, Autologous Stem cell Transplantation International Rheumatoid Arthritis Trial) started in 2003. A STUDY PROJECT IS PLANNED FOR MULTIPLE SCLEROSIS (Astims, Autologous Stem cell Transplantation International Multiple Sclerosis). PMID- 15798554 TI - [Guidelines for the medical follow-up of adult and adolescent asthmatic patients]. PMID- 15798555 TI - [Misleading pulmonary opacities]. PMID- 15798556 TI - [Panorama/In the literature When the thyroid is at the heart of debates]. PMID- 15798557 TI - [Cutaneous pathomimia: in favor of its management in dermatology without direct confrontation]. PMID- 15798558 TI - [Therapeutic training, a major stake in dermatology]. PMID- 15798559 TI - [Bullous pemphigoid in young patients: a retrospective study of 74 cases]. AB - INTRODUCTION: Bullous pemphigoid usually affects elderly people. Only a few isolated cases among people younger than 65 years have been reported. OBJECTIVES: Describe the clinical and biological characteristics of patients younger than 60 years suffering from bullous pemphigoid, compare them with the usual characteristics known among elderly people and search for potential pathological associations. PATIENTS AND METHODS: Retrospective, national, multicenter study. Clinical, biological and histological characteristics were recorded with a standardised questionnaire as well as treatments and associated pathologies. RESULTS: Seventy-four cases of bullous pemphigoid diagnosed between June 1970 and March 2002 were analyzed. Mean age at the beginning of the disease was 46 +/- 11.6 years. Further explorations by indirect immunofluorescence of separated skin and/or immuno-electron microscopy and/or immunoblotting were performed for 42 patients (56.8 p. 100). Clinical characteristics among this restricted population were comparable to those found among the 32 other cases. Compared to usual data on bullous pemphigoid in elderly people, we observed a greater proportion of extensive form of disease (75 p. 100), a more frequent head and neck involvement (39.2 p. 100) and an overexpression of anti-BP180 autoantibodies (48 p. 100). Neoplasm was notified for 7 patients (9.5 p. 100), 18 (24.3 p. 100) suffered from a pathology of the basement membrane zone (6 psoriasis, 6 atopic dermatitis and 6 lichen) and 13 from neurological disease, among which 4 were bedridden. Fourty six patients (62.2 p. 100) received drugs for the long term (mean 2.12 +/- 2.43), 4 patients were treated by PUVAtherapy and 2 by radiotherapy. DISCUSSION: Our results suggest that bullous pemphigoid among young people is more severe and more active than the usual form in the elderly. This particular form could be the result of a higher expression of anti-BP180 autoantibodies, which are considered as a marker of poor prognosis in this disease. We also found a high frequency of pathological associations and physical treatment, all responsible for damage to the basement membrane zone, which can involve auto-immunization against hemidesmosome components. PMID- 15798560 TI - [Vulvar contact dermatitis due to seminal allergy: 3 cases]. AB - BACKGROUND: Contact sensitivity deserves to be assessed in acute vulvitis as well as chronic vulvar dermatitis with a specific orientation toward semen allergy whenever the pathology is post-coital. OBSERVATIONS: Three observations of semen allergy are reported. Two women presented with post-coital vulvitis due to delayed hypersensitivity to semen proteins without associated specific IgE. The type 2 allergy was confirmed by positive patch-tests and disappearance of symptoms when using condoms. Another woman presented with oedematous vulvitis associated with asthma and malaise per and post-sexual intercourse. Positive prick-tests with husband's semen and blood tests for specific IgE led to the diagnosis of type 1 hypersensitivity to semen, with disappearance of both vulvar and systemic symptoms with the use of condoms. DISCUSSION: Immediate or delayed type hypersensitivity may be discovered using prick and patch tests respectively. The observation of vulvitis or vulvar discomfort should promote such investigations, whenever a post-coital chronology is suggested. The good clinical pertinence of positive skin tests must be emphasized. Although type I hypersensitivity to semen is documented in the literature, with easy detection of specific IgE, delayed type 2 allergy presents as an as yet under-diagnosed etiology of post-coital vulvitis. Patch tests may help to confirm that contact dermatitis may be due to seminal proteins. PMID- 15798561 TI - [Severe psoriasis: 160 cases]. AB - INTRODUCTION: Psoriasis is a frequent dermatosis, its prevalence is estimated of between 1 and 3 p. 100. The severe forms may threaten the functional and life prognosis of patients. PATIENTS AND METHODS: We conducted a retrospective study on 160 cases of severe psoriasis collected between 1990 and 2001. We included patients exhibiting severe psoriasis: pustular, erythrodermic or arthropathic psoriasis, the generalized forms that had developed for more than 6 months, without positive response to treatment and the forms with a PASI greater than 50. RESULTS: The patients' ages ranged from 6 to 85 years with a clear male predominance (96 men, 64 women). Nine familial cases were collected. Erythroderma was noted in 87 cases, pustular psoriasis in 31, generalized psoriasis in 31 and psoriatic rheumatism in 15 (4 of which were associated with pustular psoriasis). Local treatment with topical corticosteroids or a Vitamin D derivative was recommended in respectively 41.8 and 13.75 p. 100 of cases. The administration of general treated relied on methotrexate and was required in 68.75 p. 100 of cases. Progression was usually good, relapses were often noted when treatment was stopped. Four patients died. DISCUSSION: Although the diagnosis of psoriasis is often easy, the severe forms are still difficult to treat. Treatment is complicated, sometimes disappointing and often necessitates heavy and aggressive treatments that require strict surveillance. PMID- 15798562 TI - [Nodules on localized scleroderma or morphea]. AB - BACKGROUND: Localized scleroderma or morphea usually appears as flat or depressed lesions. OBSERVATIONS: We report 3 cases of morphea with atypical appearance, alternating pigmented and depigmented patches with nodules or sclerous bands, occurring in adult men. DISCUSSION: The occurrence of nodular elements on generalized or localized scleroderma, although rare, was first reported in the literature by Addisson in 1884. These nodules usually appear during evolution. These scleroderma are then described as being keloidal or nodular. We report 3 cases of nodules on localized scleroderma which appeared at the beginning of the dermatosis and where the scleroderma had a similar unusual irregularly pigmented appearance. PMID- 15798563 TI - [Disseminated histoplasmosis: an atypical ulcerous form in an HIV-infected patient]. AB - INTRODUCTION: Histoplasma capsulatum var capsulatum is a dimorphic fungi predominating on the American continent. It is responsible for disseminated histoplasmosis associated with AIDS. The presentation in the form of cutaneous ulceration is uncommon and misleading. OBSERVATION: A 25 year-old man presented with 3 ulcerations, of 2 to 4 cm in diameter, localized on the lower lip and knees. The patient exhibited fever, alteration in his general status of health and a pulmonary interstitial syndrome. He was seropositive for the human immunodeficiency syndrome (HIV). His lymphocyte CD4+ level was of 1/mm3. Diagnosis of histoplasmosis was established by direct examination and culture of the cutaneous ulcerations and bronchoalveolar washing fluid. DISCUSSION: The clinical aspect of cutaneous localizations of disseminated histoplasmosis is usually multiple, disseminated, papular or nodular-type lesions. Ulcerations represent less than 20% of the cases described. In our patient, the aspect of the lesions at first evoked cutaneous leishmaniosis. Direct mycological examination followed by culture confirmed the final diagnosis. PMID- 15798564 TI - [Cutaneous-mucosal paracoccidio-domycosis: the first case diagnosed in French Guiana]. AB - INTRODUCTION: Paracoccidio-domycosisis a deep mycosis due to a dimorphic fungus:Paracoccidioides brasiliensis. The principle endemic country is Brazil. We describe the first case of paracoccidio-domycosis, in its cutaneous-mucosal form, diagnosed in French Guiana. OBSERVATION: A 20 year-old Brazilian man, having mover to French Guiana a few months earlier, presented with multiple disseminated cutaneous lesions, predominating on the face, and composed of multiple nodules and two ulcerations. The clinical examination also revealed voluminous superficial lymph nodes and ulcerations of the pharynx and larynx. Direct examination, anatomopathology and culture of cutaneous biopsies revealed specific images of Paracoccidioides brasiliensis. HIV serology was negative. Treatment combining cotrimoxazole and itraconazole eliminated the lesions in one month. DISCUSSION: Because the patient had just moved to Guiana, this observation probably corresponded to an imported disease. The principle differential diagnosis was leishmaniosis. PMID- 15798566 TI - [Connubial contact dermatitis to an inhaled corticosteroid]. AB - BACKGROUND: Inhaled corticosteroids are widely used in allergic asthma and rhinitis. They are most often used alone or sometimes in association. Allergic side-effects of inhaled corticosteroids are less frequent than those of topical corticosteroids. We report a case of a connubial dermatitis to a budesonide spray. OBSERVATION: A 3-year old boy was treated for asthma by budesonide (Pulmicort) and terbutaline (Bricanyl) aerosols with an inhalation chamber (Babyhaler). From the fourth day of treatment onwards, his mother had swollen and itchy lesions on the face with conjunctivitis several hours after the administration of the corticosteroids using the inhalation chamber. The last eruptions were marked by extensive lesions. The patient reported a worsening of her eruption when she was treated with a desonide cream (Tridesonit). Prick-tests conducted later on confirmed the contact allergy to budesonide and Pulmicort spray. They were also positive for Tridesonit cream and triamcinolone acetonide. Repeated open application tests with a 17-butyrate hydrocortisone cream (Locoid) for three weeks remainded negative. DISCUSSION: Our observation is original: allergic contact dermatitis to inhaled corticosteroids is rare, the clinical presentation mimicked angioedema although it was a delayed-type hypersensitivity, hypersensitivity was limited to group B corticosteroids and it was in fact a connubial contact dermatitis. PMID- 15798565 TI - [Pellagra and panniculitis induced by chronic bacterial colonisation of the small intestine]. AB - INTRODUCTION: Pellagra is an exceptional disorder in France. The classical description of pellagra associates a photoinduced rash with neurological impairment and intestinal dysfunction. Without adapted treatment, the progression is fatal. CASE REPORT: A 62 year-old women developed a photoinduced rash, composed of circular and erythematous elements with pustular edges. She also had panniculitis, peripheral neuropathy, depressive mood and diarrhea. Her medical past was marked by epilepsy treated with sodium valproate and hydantoin. Biological exams revealed lowered plasma levels of vitamins PP, B1, B6 and zinc, secondary to intestinal impairment induced by bacteria proliferating in the small intestine. The rash resolved with vitamin PP and zinc supplementation. The bacterial colonisation was improved by long-term, sequential antibiotics. DISCUSSION: We report a rare clinical form of pellagroid rash. The rash was induced by chronic malabsorption resulting from excessive bacterial proliferation in the diverticules of the small intestine. The antiepileptic treatment could have facilitated vitamin PP et zinc deficiency. Panniculitis was related to the bacterial proliferation. We discuss in this paper the relationship between some vitamin deficiencies, their clinical manifestations and the direct role of intestinal bacterial proliferation in the cutaneous manifestations. PMID- 15798567 TI - [Polycythemia vera revealed by Wells syndrome]. AB - BACKGROUND: Wells syndrome is an eosinophilic dermatosis that is mainly reported in association with infections or insect bites, and more rarely during haematological disorders. CASE REPORT: A 32 year-old woman presented several recurrent episodes of massive swelling and erythema papulo-nodular inflammatory plaques on the buttocks that spontaneously resolved. The biopsy revealed marked infiltration of the dermis with eosinophils suggesting Wells syndrome. Examination found an isolated hepatosplenomegaly with true polycythemia and a myeloproliferative disorder. The diagnosis of Vaquez disease was made. DISCUSSION: Wells syndrome is a distinctive disease entity with a wide polymorphism of clinical and histological features, unspecific, and varying depending on the age of the lesions. Various triggering factors are involved, but this syndrome may also reveal hematological disorders. Cutaneous manifestations often occur before the hematological diagnosis, but are frequently misdiagnosed. Only two other cases of Wells syndrome associated with Vaquez disease have been reported. Our case report underlined the importance of systematic research for an hemopathy in Wells syndrome, especially in young patients. PMID- 15798569 TI - [Gene therapy and the skin]. PMID- 15798568 TI - [Trisomy 20 mosaicism revealed by pigmentary mosaicism of the Ito-type]. AB - INTRODUCTION: Ito hypomelanosis-type pigmentary mosaicism is characterized by congenital pigmentation disorders along Blaschko's lines. We report a case of Ito type pigmentary mosaicism associated with a congenital growth hormone deficiency having revealed trisomy 20 mosaicism. OBSERVATION: A 4 year-old boy presented with congenital pigmentation disorders. His history was marked by: inter-uterine delayed growth of unknown etiology, a dysmorphic syndrome, psychomotor retardation with speech problems, right cryptorchidia and an isolated, idiopathic, congenital growth hormone deficiency that had been treated with recombinant somatropine since the age of three. The clinical examination revealed alternating hypo and hyper-pigmented maculae with linear distribution on the limbs and in "twirls" on the trunk following Blaschko's lines. The blood karyotype was normal, the karyotype on fibroblasts of hypopigmented skin revealed trisomy 20 mosaicism. DISCUSSION: The occurrence of pigmentary mosaicism related to trisomy 20 mosaicism is exceptional. The combination of Ito hypomelanosis-type pigmentary mosaicism and delayed growth due to growth hormone deficiency has never been reported before. Our observation, unusual because of such an association, raises the question of the eventual existence of associated genes located on the chromosome 20 implied in the secretion of growth hormone and/or melanogenesis. It also underlines the interest of conducting cytogenic explorations on fibroblasts of damaged skin in the case of Ito-type pigmentary mosaicism, even if the blood karyotype is normal or in the absence of a patent phenotype abnormality. PMID- 15798570 TI - [Facial skin lesions following air bag activation]. PMID- 15798571 TI - [A simple technique for accessory salivary gland biopsy]. PMID- 15798572 TI - [Azatropin in severe atopic dermatitis: 24 cases]. PMID- 15798573 TI - [Fabry disease. Role of the dermatologist and therapeutic advance]. PMID- 15798574 TI - [Immunological mechanisms of cutaneous adverse drug reactions]. PMID- 15798575 TI - [Physical therapy in limb lymphedema]. PMID- 15798576 TI - [Auto-immune bullous dermatosis]. PMID- 15798577 TI - [Mutation in the gene encoding the alpha A regulatory subunit of cyclic dependant protein kinase in Carey's complex]. PMID- 15798578 TI - [Axillary basal cell carcinoma: a rare localization]. PMID- 15798579 TI - [Delayed contact dermatitis to corals]. PMID- 15798580 TI - [Mycetoma from Acremonium sp]. PMID- 15798581 TI - [Mirtazapin and gynecomastia-galactorrhea]. PMID- 15798582 TI - [Gelatinous bone marrow transformation in anorexia nervosa]. AB - Moderate hematologic abnormalities, like anemia or leukopenia, are frequently seen in anorexia nervosa, whereas pancytopenia and bone marrow abnormalities are uncommon. We report a case of tricytopenia with gelatinous bone marrow transformation in anorexia nervosa. Marrow gelatinous transformation (also called serous fat atrophy or starvation marrow) is characterized by the association of marrow hypoplasia and interstitial infiltration of a ground gelatinous substance (acidic mucopolysaccharides). Changes in peripheral blood cell counts are various and moderate, and do not always reflect the severity of bone marrow damage. The pathogenesis is not yet well elucidated but is certainly related to the nutritional status because gelatinous bone marrow transformation is found in anorexia nervosa and in other clinical situations with cachexia. Gelatinous transformation of the marrow is reversible with feeding. PMID- 15798584 TI - [Late puberty and/or gonadotrope insufficiency?]. PMID- 15798585 TI - [Micropolycystic ovaries and insulin-resistance syndrome]. PMID- 15798586 TI - [Steroid-secreting gonad tumors]. PMID- 15798587 TI - [Should we prescribe DHEA?]. PMID- 15798588 TI - [Update on menopause treatments, after recent controversies]. PMID- 15798589 TI - [New treatments for erectile dysfunction: an endocronologist's point of view]. PMID- 15798592 TI - [Current developments in lipid-lowering treatments]. PMID- 15798593 TI - [Type 2 diabetes in young subjects]. PMID- 15798594 TI - [Can type 2 diabetes be predicted?]. PMID- 15798595 TI - [Ultimate progress in therapeutic education: ambulatory treatment self-teaching]. PMID- 15798596 TI - [Pathology of the pituitary stalk and diabetes insipidis]. PMID- 15798597 TI - [Medical treatment of acromegaly]. PMID- 15798598 TI - [Treatment of somatotrope deficiency in adults]. PMID- 15798600 TI - [Primary hyperparathyroidism: before and during surgery]. PMID- 15798599 TI - [Endocrine tumors of the pancreas]. PMID- 15798601 TI - [Paraneoplastic hormone secreting syndromes]. PMID- 15798603 TI - [Autoimmune polyendocrinopathies]. PMID- 15798605 TI - [Papillary and/or vesicular thyroid cancer: before and during surgery]. PMID- 15798606 TI - [Monitoring papillary and vesicular cancers]. PMID- 15798607 TI - Cerebrospinal fluid MR dynamics and risk of falls in the elderly. AB - RATIONALE AND OBJECTIVES: To investigate the relationship between CSF dynamics and risk of falls of unknown origin in the elderly. POPULATION AND METHODS: Phase contrast MR studies allowed CSF aqueductal flow quantification on 23 community dwelling older people initially explored for mild cognitive impairment. Mobility assessment included report of falls, talking walking test, stance test, one leg standing test, up and go test, and measurement of fast gait speed. RESULTS: History of falls was associated with larger aqueduct, steeper diastolic slopes higher ratios RDV/SD of diastolic volume/CSF systole duration (p 0.05). The HGV infection seems to distribute in all ethnicities all over the world, implying the global importance of this hepatitis virus infection. Screening for HGV RNA might be an important test in blood bank process in the future. PMID- 15798661 TI - Histological spectrum of liver in HIV - autopsy study. AB - INTRODUCTION: Liver abnormalities are common in HIV positive patients. They are usually part of generalized process and rarely produce significant liver failure. AIM: To evaluate histological spectrum of liver disease in HIV positive patients and to ascertain if any pathologic features are widespread among HIV infection. MATERIAL & METHODS: Autopsy data from year 1991 to 2003 consisting of 60 HIV positive patients were evaluated. Demographic profile, clinical and laboratory data were obtained from hospital records. Macroscopic findings of all organs at autopsy were noted. Histological features of liver were studied in detail using routine H & E., Ziehl-Neelson stain for acid fast bacilli and other special stains such as PAS and GMS for fungal infection were done whenever indicated. RESULTS: Patients were in age group 19 to 55 years with mean age of 32.1 year; male to female ratio was 4:1. Evidence of tuberculosis either pulmonary or extrapulmonary was found in 35 (58.3%) cases. On histological examination of liver, tubercular granulomas were seen in 19 (31.6%) cases of disseminated tuberculosis. Granulomas were typical caseating epitheloid cell type in 14 (73.6%) and in 5 cases granulomas were not typical. Acid fast bacilli were demonstrated in 4 (6.6%) cases, all of which showed presence of granulomas. Other histological findings were sinusoidal and centrivenular congestion in 14 (23.3%), extensive fatty change 6 (10%), portal inflammation resembling chronic hepatitis 5 (8.3%), focal necrosis 2 (3.3%), Kupffer cell hyperplasia 1 (1.6%) and metastasis from known case of adenocarcinoma of pancreas 1 (1.6%). Associated hepatitis C and B infections were seen in 4 (6.6%) and 1 (1.6%) respectively. Opportunistic infection was seen in only 1 (1.6%) case with disseminated cryptococcosis involving liver. In remaining 6 (10%) liver histology was normal. Considering cause of death, 58.3% patients died due to disseminated tuberculosis. Patients with associated hepatitis B & C infection died due to liver cirrhosis and the remaining died of miscellaneous conditions, which were not related to HIV infection. CONCLUSION: Histopathological findings of the liver in HIV positive patients were mainly non specific. Tuberculosis was the commonest infection noted. There was no significant mortality observed specifically related to liver involvement in these patients. PMID- 15798662 TI - Trends in liver disease prevalence in Mexico from 2005 to 2050 through mortality data. AB - BACKGROUND AND AIMS: The epidemiology of liver cirrhosis differs across sex, ethnic groups, and geographic regions. In 2000, chronic liver disease was the fifth leading cause of death in Mexico. Accurate knowledge of the demographics of liver disease is essential in formulating health-care policies. Our main aim was to project the trends in liver disease prevalence in Mexico from 2005 to 2050 based on mortality data. METHODS: Data on national mortality reported for the year 2002 in Mexico were analyzed. Specific-cause mortality rates were calculated for a selected age population (> 25 years old) and classified by sex and projected year (2005-2050). The following codes of the International Classification of Diseases for liver diseases were included: non-alcoholic chronic liver disease and cirrhosis, alcoholic liver disease, liver cancer, and acute and chronic hepatitis B and C infection. The projected prevalence of a chronic liver disease was estimated using the following equation: P = (ID x T) / [(ID xT) + 1], where P = prevalence, ID = incidence density (mortality rate multiplied by 2), T = median survival with the disease (= 20 years). RESULTS: Nearly two million cases of chronic liver disease are expected. Alcohol-related liver diseases remain the most important causes of chronic liver disease, accounting for 996,255 cases in 2050. An emergent syndrome is non-alcoholic liver disease, which will be more important that infectious liver diseases (823,366 vs 46,992 expected cases, respectively). Hepatocellular carcinoma will be the third leading cause of liver disease. CONCLUSIONS: Chronic liver disease will be an important cause of morbidity and mortality in the future. Preventive strategies are necessary, particularly those related to obesity and alcohol consumption, to avoid catastrophic consequences. PMID- 15798663 TI - Combination of Peginterferon alpha-2b (12 kDa) and Lamivudine in difficult-to treat chronic hepatitis B- an Indian experience. AB - INTRODUCTION: Treatment strategies for chronic hepatitis B (CHB) using either interferon or Lamivudine can achieve sustained response in 30-40 % in HBeAg positive CHB. Retreatment of treatment failure in CHB and treatment of HBeAg negative patients pose major therapeutic challenge. Because of success story of Peginterferon in hepatitis C and some preliminary data in CHB, we undertook this open-labeled prospective study to study the response of Peginterferon alpha-2b (PegIFN) and Lamivudine combination in patients of CHB. MATERIALS AND METHODS: Following 4 groups of patients of CHB with persistently elevated transaminases were treated with Lamivudine 100 mg PO daily and PegIFN 1.5 microg/kg SC once a week- 1) HBeAg negative treatment naive patients- for 12 months 2) HBeAg negative patients who were nonresponsive to at least two treatment regimens for 12 months, 3) HBeAg positive treatment naive patients- for 6 months 4) HBeAg positive patients who were treatment failure at least with two regimens- for 6 months. Patients were tested for LFT, HBeAg, antiHBe, quantitative HBVDNA (only in HBeAg negative CHB) and liver biopsy when possible at inclusion. During treatment period, LFT was tested at monthly interval and HBeAg, antiHBe and HBVDNA (in HBeAg negative CHB) at 3 monthly intervals. End of treatment response (EOR) was assessed at end of treatment period and sustained response (SR) at 6 months post treatment. Treatment response was defined in HBeAg positive patients with normalization of enzymes and disappearances of HBeAg & in antiHBe positive patients with normalization of enzymes and loss of detectable HBVDNA. RESULTS: Total 25 patients with CHB were included in this study with mean age of 37.9 +/- 4.6 years (range 20-56) and male: female= 11.5:1. 1) In HBeAg negative treatment naive patients, 4/6 patients achieved EOR (66.6%), whereas in 3/6 patients (50%) had SR. 2) In HBeAg negative treatment failure group, 4/5 patients had EOR (80%) and 3/5 patients achieved SR (60%), of which 1 patient lost HBsAg. 3) In HBeAg positive treatment naive patients, 4/5 patients achieved EOR (80%) and 3/5 patients obtained SR (60%). One patient in this group lost HBsAg. 4) In HBeAg positive treatment failure group, 5/9 patients achieved EOR (55.5%), whereas 3/9 patients (33.3%) obtained SR. CONCLUSION: Overall, in difficult-to-treat patients (considering groups 1, 2 & 4- total 20 patients) with combination of PegIFN and Lamivudine, EOR was seen in 13 patients (65%), SR was obtained in 9 patients (45%) and loss of HBsAg was seen in 1 patient (5%). PMID- 15798664 TI - Hepatic angiosarcoma. PMID- 15798665 TI - Infection control in the Eastern Mediterranean region: time for collaborative action. PMID- 15798666 TI - In vitro activity of antimicrobial agents against extended-spectrum beta lactamase-producing Escherichia coli and Klebsiella pneumoniae at a tertiary care center in Lebanon. AB - Looking for therapeutic options, we assessed the minimum inhibitory concentrations (MICs) of 7 antimicrobial agents against extended-spectrum beta lactamase-producing Klebsiella pneumoniae (n = 58) and Escherichia coli (n = 84) isolates. High rates of susceptibility were shown for both E coli and K pneumoniae against ertapenem (100% for both), piperacillin/tazobactam (83% and 91%, respectively) and amikacin (96% and 82%, respectively). In addition, most K pneumoniae isolates were susceptible to quinolones (72%-75%) and cefepime (88%). However, clinical correlation is warranted. PMID- 15798667 TI - Ventriculostomy-associated infections: incidence and risk factors. AB - BACKGROUND: The objective of this study was to assess the incidence of ventriculostomy-associated infections (VAI) and to examine the related risk factors. METHODS: Data on all consecutive patients with ventriculostomy catheters admitted to the intensive care unit (ICU) in a tertiary care center over a 3-year period were identified from the ICU database and from medical records. VAI was documented using a preset definition. The following patient data were documented: demographics, severity of illness measures, indication for the catheter, presence of cerebrospinal fluid (CSF) leak, and length of stay and mortality. The following catheter data were collected: venue of catheter placement (operating room or nonoperating room areas), use of antibiotic irrigation and prophylactic systemic antibiotics, and number of catheter days. The frequency of CSF sampling was documented. RESULTS: In 84 patients, 99 catheters were placed, of which 19% developed VAI. There was a total of 586 catheter days (infection rate, 32 per 1000 catheter days). The risk of VAI increased steadily until catheter day 7 then reached a plateau. Among patients' factors, repeat catheter insertion was associated with a significant increase in VAI. There were no significant associations with age, severity of illness, indication for the catheter, craniatomy, or presence of CSF leak. Among catheter factors, the number of catheter days and repeat catheter insertion emerged as significant independent predictors on multivariate analysis. Placement outside the operating room was associated with a trend toward higher VAI. The use of prophylactic antibiotic or antibiotic irrigation did not significantly alter VAI rates. Routine surveillance cultures of CSF were no more likely to detect infection than cultures obtained when clinically indicated. Gram-negative bacilli were responsible for 50% of the infections, followed by gram-positive cocci (29%) and others (21%). CONCLUSIONS: The risk of VAI increases with increasing duration of catheterization and with repeated insertions. The use of local antibiotic irrigation or systemic antibiotics does not appear to reduce the risk of VAI. Routine surveillance cultures of CSF were no more likely to detect infection than cultures obtained when clinically indicated. These findings need to be considered in infection control policies addressing this important issue. PMID- 15798668 TI - Compliance to hepatitis B vaccination and subsequent development of seroprotection among health care workers of a tertiary care center of Saudi Arabia. AB - BACKGROUND: There is lack of reliable data on compliance to hepatitis B virus (HBV) vaccine and development of seroprotective levels of antibodies among health care workers (HCWs) from the countries with high HBV endemicity such as Saudi Arabia. This study aimed to assess the compliance with HBV vaccine and subsequent levels of seroprotection among HCWs of a large tertiary care center of the Eastern Province of Saudi Arabia. METHODS: All the HCWs (n = 1302) involved in direct patient care, including 374 (28.7%) physicians, 619 (47.6%) nurses, and 309 (23.7%) technicians, were enrolled for the study. Those having antibody to hepatitis B surface antigens (anti-HBs) levels less than 10 mIU/mL were advised to take 3 doses of yeast-derived recombinant HBV vaccine at 0, 1, and 6 months of 1 mL (20 microg/mL) in the deltoid muscle. Blood samples were checked for anti HBs antibody levels by enzyme immunoassay during the initial screening of HCWs and 3 months after the third dose of HBV vaccine. The group of physicians included 34.5% (129/374) of consultants, 16.8% (63/374) of specialists, and 48.6% (182/374) of residents. RESULTS: An overall HBV vaccine compliance rate of 71.6% (932/1302) was observed among HCWs including that of 79.5% (492/619) among nurses, 78.3% (242/309) among technicians, and 52.9% (198/374) among physicians. Thus, physicians recorded the lowest compliance (OR, 3.211; 95% CI, 2.259-4.567; P < .0001) to HBV vaccine. Among physicians, the lowest compliance of 42.3% (77/182) was observed in residents (OR, 3.690; 95% CI, 1.067-3.703; P < .0001). The overall seroprotection after vaccination was achieved in 92.2% of the compliant HCWs, and 7.8% of them failed to mount adequate response to HBV vaccine. Nonresponders included mainly the physicians (OR, 2.229; P = .05) consultants in particular (OR, 3.476; P < .0001). The mean age of nonresponders was higher than those who mounted an adequate anti-HBs response (46.7 +/- 6.3 vs. 32.2 +/- 3.3 years OR, 1.845; 95% CI, 0.999-3.414; P < .05). CONCLUSIONS: Poor compliance to HBV vaccine among physicians--residents in particular-is an issue of immense concern, which demands close examination and identification of the specific action that needs to be taken to enhance the uptake of the vaccine by this target population. PMID- 15798669 TI - The utility and interpretation of tuberculin skin tests in the Middle East. AB - Tuberculin skin test (TST) interpretation can be confounded by Bacillus Calmette Guerin (BCG) vaccination and infection with nontuberculosis mycobacteria (NTM). However, a wealth of epidemiologic information has allowed the formulation of recommendations for reasonably informed interpretation of the TST in most clinical situations. In the Middle East, the TST remains a useful test. BCG vaccination is given at birth, which should have minimal effect on TST reactions in adolescents or adults. In countries of the Middle East with moderate to high incidence of active smear-positive pulmonary TB (>20 per 100,000 per year), a positive TST will almost always indicate true TB infection. However, in Middle East countries with very low incidence of active TB (<10 per 100,000 per year), a positive TST will more likely be false positive because BCG vaccination is still routinely given; until BCG vaccination is abandoned, the TST will be less useful in these countries. These findings are applicable to countries in other regions of the world, and the utility TST will also be increased where the likelihood of TB infection is higher and lowered where TB infection is unlikely, yet BCG vaccination is still given. PMID- 15798670 TI - The attributable cost and length of hospital stay because of nosocomial pneumonia in intensive care units in 3 hospitals in Argentina: a prospective, matched analysis. AB - BACKGROUND: No information is available on the financial impact of nosocomial pneumonia in Argentina. To calculate the cost of nosocomial pneumonia in intensive care units, a 5-year, matched cohort study was undertaken at 3 hospitals in Argentina. SETTING: Six adult intensive care units (ICU). METHODS: Three hundred seven patients with nosocomial pneumonia (exposed) and 307 patients without nosocomial pneumonia (unexposed) were matched for hospital, ICU type, year admitted to study, length of stay more than 7 days, sex, age, antibiotic use, and average severity of illness score (ASIS). The patient's length of stay (LOS) in the ICU was obtained prospectively in daily rounds, the cost of a day was provided by the hospital's finance department, and the cost of antibiotics prescribed for nosocomial pneumonia was provided by the hospital's pharmacy department. RESULTS: The mean extra LOS for 307 cases (compared with controls) was 8.95 days, the mean extra antibiotic defined daily doses (DDD) was 15, the mean extra antibiotic cost was $996, the mean extra total cost was $2255, and the extra mortality was 30.3%. CONCLUSIONS: Nosocomial pneumonia results in significant patient morbidity and consumes considerable resources. In the present study, patients with nosocomial pneumonia had significant prolongation of hospitalization, cost, and a high extra mortality. The present study illustrates the potential cost savings of introducing interventions to reduce nosocomial pneumonia. To our knowledge, this is the first study evaluating this issue in Argentina. PMID- 15798671 TI - New methods in the evaluation of chemical disinfectants used in health care services. AB - BACKGROUND: A considerable number of tests are recommended in the literature to evaluate in vitro commercial chemical solutions. The variety of tests reflects their limitations and the need to enhance disinfection process. METHODS: In this study the efficacy of 4 chemical disinfectants selected by their practical use in Health Care Services and by literature recommendation in aerobic and in strict anaerobic bacteria were evaluated by their practical use in Health Care Services and by literature recommendation in aerobic and in strict anaerobic bacteria. Viability was tested in biofilms grown on glass and rubber tip carriers. RESULTS: The results showed microbial growth in chemical solutions at concentrations recommended by the literature or at very close concentrations to them. Viable cells were recovered from biofilms after 30 minutes (Bacteroides fragilis) and 60 minutes (Streptococcus mutans and Salmonella tiphymurium) contact with 2.4% glutaraldehyde and after 60 minutes (S tiphymurium) in 2.0% glutaraldehyde. In 70% ethyl alcohol, S tiphymurium was viable up to 10 minutes, Escherichia coli up to 30 minutes, and S mutans up to 60 minutes. In 1% sodium hypochlorite, S mutans was viable up to 30 minutes and S tiphymurium up to 45 minutes. Detection of cell viability could be related to methodologic differences, including biofilm formation, as demonstrated by scanning electron microscopy. It should be emphasized that B fragilis, the most clinically relevant obligate anaerobe, remained viable in one routinely used solution. CONCLUSION: These findings pointed out the need of periodic surveillance of disinfectants' activity used in Health Care Services and the need of reviewing routines of disinfection protocols. PMID- 15798672 TI - Microbial contamination of hospital bed handsets. AB - BACKGROUND: Hospital bed handsets, including nurse call equipment and television controls, have been found to contain biologic material and may be contaminated with microbes. OBJECTIVE: The aim of this study was to assess the microbial contamination of hospital bed handsets. METHODS: Hospital bed handsets were removed from 115 randomly chosen rooms in a suburban hospital. The handsets were transported to the laboratory in a sterile fashion and opened using a sterile technique, and cultures were obtained from both the anterior and posterior surfaces of the units. RESULTS: The cultures of 12 units (10.4%) revealed no microorganisms. One hundred three units (89.6%) had cultures that grew microorganisms. Of the handsets that were found to contain microorganisms, 48 units (46.6%) had only 1 microorganism, and 55 units (53.4%) had multiple organisms, including 33 units (32.0%) with 2 microorganisms, 21 units (20.4%) with 3 microorganisms, and 1 unit (1.0%) with 4 microorganisms. The microorganisms identified included 90 isolates (87.4%) of coagulase-negative staphylococcus, 51 isolates (49.5%) of bacillus species, 13 isolates (12.6%) of fungal species, 8 isolates (7.8%) of nonhemolytic streptococcus species, 7 isolates (6.8%) of alpha-hemolytic streptococcus species, 1 isolate (1.0%) of Staphylococcus aureus, and 1 isolate (1.0%) of methicillin-resistant Staphylococcus aureus. CONCLUSION: Hospital bed handsets were found to have a high incidence of contamination with bacteria and fungus and were found to contain organisms that are known to be the etiologic agents in nosocomial infections. Because of the frequency and duration of contact between hospital patients and hospital bed handsets, existing infection control measures should be studied that could reduce the level of contamination of such handsets or that could isolate the handsets from the patient. PMID- 15798673 TI - Assessing motivation for physicians to prevent antimicrobial resistance in hospitalized children using the Health Belief Model as a framework. AB - BACKGROUND: Antimicrobial resistance (AR) is a threat to hospitalized children, and more information is needed to motivate physicians to adopt evidence-based practices such as those in the Centers for Disease Control and Prevention's (CDC) Campaign to Prevent Antimicrobial Resistance. METHODS: The framework of the Health Belief Model (HBM) was used to assess personal perceptions and cues to action related to the prevention of AR in 3 national focus groups in August 2003. RESULTS: Twenty-five physicians who treat hospitalized children participated; all reported that they had cared for a patient with an antimicrobial-resistant organism. Physicians perceived that AR was more of a problem nationally (92%) than in their institution (76%) or practice (60%) and reported that issues such as nursing shortage, cost of health care, and lack of specialty services presented more of a challenge to the care of hospitalized children than AR. Reported preferences of cues to action included journal articles (80%), infectious disease experts (76%), and colleagues (52%). CONCLUSIONS: The HBM provides insight into physicians' perceptions about AR and preferred cues to action, which yield valuable information concerning the modes, methods, and messengers to intervene on problems such as antimicrobial resistance in hospitalized children. PMID- 15798674 TI - A National Guard outbreak of Burkholderia cepacia infection and colonization secondary to intrinsic contamination of albuterol nebulization solution. AB - BACKGROUND: Burkholderia cepacia, a gram-negative pathogen, has been a known cause of hospital outbreaks because of a contaminated common source such as multidose medications. We describe an outbreak with Burkholderia cepacia infection in 2 major hospitals affiliated to the National Guard, related to an intrinsic contamination of a locally manufactured, multidose Albuterol nebulization solution (Tabouk Pharmaceutical Company, Tabouk, Saudi Arabia) and we report the interventions taken to interrupt this outbreak. METHODS: During the outbreak period between May 2003 and March 2004, a combined prospective surveillance and a retrospective chart and microbiologic data review were conducted in 4 major hospitals affiliated to the National Guard. Microbiologic cultures were also performed on environmental objects of concern, as well as certain medications. In addition, a questionnaire was distributed to the respiratory therapy staff to evaluate the process of administering respiratory medications and their adherence to sound infection control practices. RESULTS: An intrinsic contamination of a locally manufactured brand of multidose Albuterol nebulization with B cepacia was identified. Two of the 4 hospitals were found to be involved: hospital A a 700-bed tertiary care center and Hospital B a 150-bed hospital. A total of 2121 patients were exposed to Albuterol nebulization as inpatients at hospital A and 318 as outpatients. For hospital B, a total of 283 inpatients and 34 outpatients were exposed to the Albuterol nebulization. Forty and 12 patients, from hospital A and hospital B, respectively, were found to have at least 1 positive culture for B cepacia. From hospital A, most samples were respiratory, and, from hospital B, most were from blood. Molecular typing of 34 available isolates showed that 23 cases were of a single strain of B cepacia that matched the strain isolated from the 3 different batches of multidose Albuterol nebulization. Three culture-positive patients never received Albuterol nebulization of that brand but were in the same room of a patient who had been receiving the medication. CONCLUSIONS: We identified a large outbreak of B cepacia in 2 major hospitals affiliated with the National Guard, linked to an intrinsic contamination of a multidose Albuterol nebulization solution. During the period of prospective surveillance, only a few cases were identified as a result of nosocomial transmission. Immediate notification of the Ministry of Health and withdrawal of the medication and revisiting the respiratory therapy practices were necessary to halt this outbreak. PMID- 15798676 TI - Saudi Arabia-United States collaboration in health research: a formula for success. AB - The aim of this article is to share our experiences from an international collaborative effort to study health outcomes among Saudi Arabian National Guard (SANG) soldiers following the 1991 Gulf War. By paying particular attention to distinct social and religious customs, geopolitical differences, and unique aspects of the health care system, we achieved a successful international collaboration in health research. PMID- 15798675 TI - Low prevalence of antibodies to human T-lymphotropic virus-I/II among blood donors in eastern Saudi Arabia. AB - The seroprevalence of human T-lymphotropic virus (HTLV)-I/II was assessed in 13,443 consecutive blood donors in eastern Saudi Arabia between 1998 and 2001. Screening by enzyme-linked immunosorbent assay (ELISA) and confirmation by Western blot resulted in 8 (0.060%) positive cases, of which 5 (0.056%) belonged to Saudi and 3 (0.113%) to non-Saudi donors. The majority of the HTLV-positive donations (6/8) were for patients, and none had a history of known risk factor for HTLV-I/II transmission. Although the very low prevalence of HTLV-I/II among Saudi donors does not support routine screening, screening of donors from other nationalities may be initiated, especially those from HTLV-I/II endemic areas. PMID- 15798677 TI - Factors influencing centric relation records in edentulous mouths. 1964. PMID- 15798678 TI - The Journal of Prosthetic Dentistry publication standards of ethical conduct. PMID- 15798679 TI - The use of porcelain laminate veneers and a removable gingival prosthesis for a periodontally compromised patient: a clinical report. AB - Periodontal disease may lead to tooth and tissue loss that can result in esthetic problems. Combined periodontal/prosthodontic treatment for patients with advanced periodontal disease is well documented. This clinical report illustrates a method of treatment for advanced tissue loss in an esthetic area using porcelain laminate veneers and a removable gingival prosthesis. PMID- 15798681 TI - An indirect repair technique for fractured metal-ceramic restorations: a clinical report. AB - Metal-ceramic crowns and fixed partial dentures have the potential for fracture of the ceramic veneer. Fractured porcelain poses a serious cosmetic and clinical problem. It may be desirable to repair a broken retainer of a fixed prosthesis rather than to remove the prosthesis and risk the possibility of destroying the entire restoration or damaging the abutment teeth. This article presents an indirect method that may be used to repair metal-ceramic restorations that are otherwise clinically and radiographically acceptable. With this technique, a new metal-ceramic crown is fabricated and is cemented on the prepared fractured retainer of the prosthesis. PMID- 15798680 TI - Contact stomatitis due to palladium in dental alloys: a clinical report. AB - A patient was treated with a maxillary gold-palladium alloy fixed partial denture (FPD). Shortly after placement, the patient contracted severe contact mucositis. Patch-testing revealed a positive reaction to palladium chloride (PdCl 2 , 1% petrolatum) but not to any other component of the prosthesis. After removal of the FPD and placement of a provisional acrylic resin restoration, all signs and symptoms disappeared. A definitive metal-ceramic (titanium-porcelain) prosthesis was placed, and no signs of lesions appeared. This clinical report demonstrates that titanium may be a satisfactory alternative for patients who require prostheses and are sensitive to other metals. PMID- 15798682 TI - Static implant loading caused by as-cast metal and ceramic-veneered superstructures. AB - STATEMENT OF PROBLEM: The passive fit of superstructures for implant-supported restorations is affected by each step of the fabrication process. In this context the question arises whether ceramic veneering would increase static implant loading. PURPOSE: The purpose of this study was to quantify the strain development of various fixed partial dentures (FPDs) both in the as-cast condition and after ceramic veneering. MATERIAL AND METHODS: Four different types (n = 10) of 5-unit FPDs (cementable, screw retained/plastic cylinder, screw retained/gold cylinder, screw retained/cemented) representing commonly used types of FPDs were investigated before and after ceramic veneering. Three implants were placed in a model simulating a patient situation, and strain gauges were mounted mesially and distally adjacent to the implants. The strain development was recorded during cement setting (provisional cement) and screw fixation. The data were analyzed statistically using multivariate 2-sample tests (alpha=.1). RESULTS: All FPDs revealed measurable amounts of strain. Neither the type of retention nor the mode of fabrication for conventional screw-retained FPDs had a significant influence on strain development. Ceramic veneering caused an increase in strain development for the conventional fixed partial dentures tested. The lowest strains were found in FPDs cemented to gold cylinders on the model for the metal frames and the ceramic-veneered FPDs. CONCLUSION: Conventional procedures were unable to produce superstructures with absolute passive fit. Ceramic veneering appeared to increase strain development and, thus, inaccuracy of the fit. The technique of cementing superstructures to prefabricated components directly on the implants may compensate for dimensional errors caused by impression making and superstructure fabrication. PMID- 15798683 TI - In vitro fracture resistance of endodontically treated central incisors with varying ferrule heights and configurations. AB - STATEMENT OF PROBLEM: The in vitro effectiveness of a uniform circumferential ferrule has been established in the literature; however, the effect of a nonuniform circumferential ferrule height on fracture resistance is unknown. PURPOSE: This in vitro study investigated the resistance to static loading of endodontically treated teeth with uniform and nonuniform ferrule configurations. MATERIAL AND METHODS: Fifty extracted intact maxillary human central incisors were randomly assigned to 1 of 5 groups: CRN, no root canal treatment (RCT), restored with a crown; RCT/CRN, no dowel/core, restored with a crown; 2 FRL, 2-mm ferrule, cast dowel/core and crown; 0.5/2 FRL, nonuniform ferrule (2 mm buccal and lingual, 0.5 mm proximal), cast dowel/core and crown; and 0 FRL, no ferrule, cast dowel/core and crown. The teeth were prepared to standardized specifications and stored for 72 hours in 100% humidity prior to testing. Testing was conducted with a universal testing machine with the application of a static load, and the load (N) at failure was recorded. Statistical analysis was performed with a 1-way analysis of variance and the Tukey Honestly Significant Difference test (alpha=.05). The mode of fracture was noted by visual inspection for all specimens. RESULTS: There was strong evidence of group differences in mean fracture strength ( P <.0001). Following adjustment for all pairwise group comparisons, it was found that the lack of a ferrule resulted in a significantly lower mean fracture strength (0 FRL: 264.93 +/- 78.33 N) relative to all other groups. The presence of a nonuniform (0.5 to 2-mm vertical height) ferrule (0.5/2 FRL: 426.64 +/- 88.33 N) resulted in a significant decrease ( P =.0001) in mean fracture strength when compared with the uniform 2-mm vertical ferrule (2 FRL: 587.23 +/- 110.25 N), the group without RCT (CRN: 583.67 +/- 86.09 N), and the RCT-treated tooth with a crown alone (CRN/RCT: 571.04 +/- 154.86 N). The predominant mode of failure was an oblique fracture extending from the lingual margin to the facial surface just below the insertion of the tooth into the acrylic resin. CONCLUSION: The results demonstrated that central incisors restored with cast dowel/core and crowns with a 2-mm uniform ferrule were more fracture resistant compared to central incisors with nonuniform (0.5 to 2 mm) ferrule heights. Both the 2-mm ferrule and nonuniform ferrule groups were more fracture resistant than the group that lacked a ferrule. PMID- 15798684 TI - The effect of box preparation on the strength of glass fiber-reinforced composite inlay-retained fixed partial dentures. AB - STATEMENT OF PROBLEM: Nonstandardized box dimensions for inlay-retained fixed partial dentures (FPDs) may result in uneven distribution of the forces on the connector region of such restorations. PURPOSE: The objective of this in vitro study was to evaluate the effect of box dimensions on the initial and final failure strength of inlay-retained fiber-reinforced composite (FRC) FPDs. MATERIAL AND METHODS: Twenty-one inlay-retained FPDs were prepared using FRC (everStick) frameworks with unidirectional fiber reinforcement between mandibular first premolars and first molars. Boxes were prepared using conventional inlay burs (Cerinlay), and small and large ultrasonic tips (SONICSYS approx). Box dimensions were measured after preparation with a digital micrometer. All restorations were subjected to thermal cycling (6000 cycles, 5 degrees C-55 degrees C). Fracture testing was performed in a universal testing machine (1 mm/min). Acoustic emission signals were monitored during loading of the specimens. Initial and final fracture strength values (2-way ANOVA, Bonferroni post hoc tests, alpha =.05) and failure types (Fisher exact test) were statistically compared for each group. RESULTS: Significant differences (P =.0146 and P =.0086) were observed between the groups in the dimensions of the boxes prepared using conventional burs buccolingually (2.8-3.0 mm in molars, 3.1-4.3 mm in premolars) and the small size (2.5-2.9, 2.9-3.8 mm) or large size (2.6-3.8, 3.2-4.9 mm) ultrasonic tips for the premolars and the molars, respectively. No significant differences were found at the initial and final failures between the conventionally prepared group (842 +/- 267 N, 1161 +/- 428 N) and those prepared with either small (1088 +/- 381 N, 1320 +/- 380 N) or large ultrasonic tips (1070 +/- 280 N, 1557 +/- 321 N), respectively. The failure analysis demonstrated no significant difference in failure types but predominant delamination of the veneering resin (85%) in all experimental groups. According to acoustic emission tests, a higher energy level was required for final failure of the FRC FPDs with boxes finished using small ultrasonic tips. CONCLUSION: Standardized box dimensions showed no significant effect on fracture strength at either initial or final failure of the fiber-reinforced FPDs. The FRC FPDs with boxes refined with small ultrasonic burs required a greater energy level before failure. The type of failure observed after the fracture tests was primarily delamination of the veneering resin. PMID- 15798685 TI - Influence of firing cycles on the margin distortion of 3 all-ceramic crown systems. AB - STATEMENT OF PROBLEM: Although all-ceramic restorations are widely used, there is a lack of information about how the fit is affected by fabrication procedures. The adequacy of the fit of all-ceramic restorations has been questioned. PURPOSE: This study examined the effect of porcelain and glaze firing cycles on the fit of 3 types of all-ceramic crowns. MATERIAL AND METHODS: Ten standardized all-ceramic crowns were fabricated on a metal die from each of 3 systems: conventional In Ceram, copy-milled In-Ceram, and copy-milled feldspathic crowns. Copings of the conventional and copy-milled In-Ceram crowns and nonglazed copy-milled feldspathic crowns served as the control. A device was used to apply a uniform load on specimens during measurement and to reposition the specimens on the measurement device after each manufacturing process. The specimens were not cemented and were measured on the metal die using a profile projector. Measurements were recorded at 18 points selected along horizontal and vertical planes. The crown systems were compared by use of the Student t test and 1-way analysis of variance (ANOVA). Data of measurements repeated at identical locations were analyzed with a multivariate repeated-measures ANOVA. The Bonferroni post hoc test was used for multiple comparisons (alpha=.05). RESULTS: The conventional In-Ceram (57 +/- 24 microm) and copy-milled In-Ceram (57 +/- 32 microm) crowns demonstrated nearly identical marginal discrepancy values, followed by the copy-milled feldspathic crowns with a mean of 17 +/- 12 microm in the vertical plane. The copy-milled In-Ceram crowns had a mean horizontal discrepancy value of -12 +/- 4 microm, followed by the copy-milled feldspathic crowns with a mean of -4 +/- 5 microm and the conventional In-Ceram crowns with a mean of -6 +/- 4 microm. Statistical analyses demonstrated no significant differences in the marginal discrepancy values among the 3 all-ceramic crown systems, except for the horizontal discrepancy values between the conventional and copy-milled In-Ceram crowns after the porcelain firing cycle. Results indicated that the addition of porcelain to the copings caused a significant change ( P <.05) in the marginal fit of the crowns, except for the fit in the horizontal plane of the conventional In-Ceram crowns. However, no significant changes occurred in the fit of the 3 all-ceramic crowns after the glaze firing cycle. There were significant differences in the marginal discrepancy values among the measurement locations ( P <.05), and the discrepancy value at each location was independent of the mean of the entire crown. CONCLUSIONS: Within the limitations of this study, it was concluded that the 3 all-ceramic crown systems demonstrated a comparable and acceptable marginal fit. The porcelain firing cycle affected the marginal fit of the all-ceramic crowns. However, the glaze firing cycle had no significant effect on fit. The conventional and copy-milled In-Ceram crowns demonstrated medial deformations at the labial and palatal surfaces that might result in occlusal displacement of the crown. PMID- 15798686 TI - Effects of glow-discharge and surfactant treatments on the wettability of vinyl polysiloxane impression materials. AB - STATEMENT OF PROBLEM: Although surfactant-modified vinyl polysiloxane (VPS) is described as hydrophilic by manufacturers, interaction with dental gypsum is relatively poor. PURPOSE: The purpose of this study was to compare and evaluate the wettability of surfactant-modified VPS after different surface treatments. MATERIAL AND METHODS: Eighty-four impression-material specimens (20 x 10 x 1 mm) were prepared from 3 different brands of VPS (Aquasil, Panasil Contact Plus, and Accuflex). Four groups (n = 7) of specimens were established according to the surface treatment method used: no treatment, topical application of 2 different surface wettability agents (Silwet Copolymer L-77 or Delar surfactant), or coating with an acrylic acid plasma in a glow-discharge reactor. Plasma treatment specimens were coated with acrylic acid in a glow-discharge system at a radio frequency of 13.56 MHz with 20-W discharge power and 5 minutes of exposure time. Wettability was measured using the contact-angle method. Values were recorded after the drop contacted the surface at 0.6 second, 30.7 seconds, and 210.8 seconds. Contact-angle mean values were compared using 3-way analysis of variance with repeated measures on 1 factor, followed by a post hoc Duncan test (alpha=.05). RESULTS: As the sphericity assumption was not satisfied, the Greenhouse-Geisser correction was used for contact-angle values. Glow-discharge treatment proved to be effective for increasing the surface energy of the VPS compared to controls at all time points studied (P <.0001). At 0.6 second, Silwet significantly decreased the contact-angle values for Aquasil and for Panasil Contact Plus (P <.0001). Delar treatment, at this time point, significantly increased the contact-angle values of GC (P <.0001) and decreased the contact angle values of Aquasil (P <.0001). At 30.7 seconds, Silwet significantly increased the wettability of Aquasil and Panasil Contact Plus ( P <.0001) but did not significantly increase the wettability of GC. Delar treatment, at 30.7 seconds, increased the contact-angle values of GC (P <.0001), whereas contact angle values of Aquasil significantly decreased (P <.0001). At 210.8 seconds, all surface-modification methods used were effective in increasing the wettability of VPS tested over untreated controls (P <.0001). CONCLUSION: Wettability of VPS based impression materials can be increased by coating the surface with acrylic acid in a glow-discharge reactor. The change in wettability achieved by the application of topical surfactants resulted in variations among surfactant-VPS combinations. PMID- 15798688 TI - Flexural strength and moduli of hypoallergenic denture base materials. AB - STATEMENT OF PROBLEM: Hypoallergenic denture base materials show no residual methyl methacrylate (MMA) or significantly lower residual MMA monomer content compared to polymethyl methacrylate-based (PMMA) heat-polymerizing acrylic resin. There is insufficient knowledge of the mechanical properties of hypoallergenic denture base materials to warrant their use in place of PMMA-based acrylic resins for patients with allergic reaction to MMA. PURPOSE: This in vitro study compared flexural strength and flexural modulus of 4 hypoallergenic denture base materials with flexural strength/modulus of a PMMA heat-polymerizing acrylic resin. MATERIAL AND METHODS: The following denture base resins were examined: Sinomer (heat-polymerized, modified methacrylate), Polyan (thermoplastic, modified methacrylate), Promysan (thermoplastic, enterephthalate-based), Microbase (microwave-polymerized, polyurethane-based), and Paladon 65 (heat-polymerized, methacrylate, control group). Specimens of each material were tested for flexural strength and flexural modulus (MPa, n = 5) according to ISO 1567:1999. The data were analyzed with 1-way analysis of variance and the Bonferroni-Dunn multiple comparisons post hoc analysis for each test variable (alpha=.05). RESULTS: Flexural strength of Microbase (67.2 +/- 5.3 MPa) was significantly lower than Paladon 65 (78.6 +/- 5.5 MPa, P <.0001). Flexural strength of Polyan (79.7 +/- 4.2 MPa, P =.599), Promysan (83.5 +/- 3.8 MPa, P =.412), and Sinomer (72.3 +/- 2.1 MPa, P =.015) did not differ significantly from the control group. Significantly lower flexural modulus was obtained from Sinomer (1720 +/- 30 MPa, P =.0007) compared to the PMMA control group (2050 +/- 40 MPa), whereas the flexural modulus of Promysan (2350 +/- 170 MPa, P =.0005) was significantly higher than the PMMA material. Microbase (2100 +/- 210 MPa, P =.373) and Polyan (2070 +/- 60 MPa, P =.577) exhibited flexural modulus similar to the PMMA material. The tested denture base materials fulfilled the requirements regarding flexural strength (>65 MPa). With the exception of Sinomer, the tested denture base resins passed the requirements of ISO 1567 regarding flexural modulus (>2000 MPa). CONCLUSION: Flexural modulus of Promysan was significantly higher than the PMMA material. Microbase and Sinomer exhibited significantly lower flexural strength and flexural modulus, respectively, than PMMA. The other groups did not differ significantly from the control group. PMID- 15798687 TI - Antibacterial properties of aged dental cements evaluated by direct-contact and agar diffusion tests. AB - STATEMENT OF PROBLEM: Since failure of fixed partial dentures is most frequently caused by caries, it would be advantageous if cements possessed antibacterial properties. PURPOSE: The purpose of this study was to evaluate the antibacterial properties of 3 dental cements using the direct-contact test and agar diffusion test. MATERIAL AND METHODS: For the direct-contact test, wells (n = 4) of microtiter plates were coated with the tested cements (Harvard cement, Duralon, and Ketac-Cem) while Streptococcus mutans suspension was placed directly on the cements. Bacterial growth was evaluated by a temperature-controlled microplate spectrophotometer. Eight wells of bacteria without the tested cements served as the positive control. Six wells of the tested cement without bacteria served as the negative control. For the agar diffusion test, triplicate specimens of freshly mixed cements were poured into uniform wells (5 mm in diameter) punched in the agar plates inoculated with Streptococcus mutans . After incubation at 37 degrees C for 24 hours, the agar plates were examined for bacterial growth and the diameter of the halo formed in the bacterial lawn was measured. In both tests, each cement was mixed in 2 different powder/liquid ratios. For the direct contact test, data were initially recorded after 1 hour of incubation. Additional experiments were performed on specimens that were aged for 24 hours, 1 week, 1 month, and 3 months before assessment by either direct-contact test or agar diffusion test. The data were subjected to 1-way ANOVA with the Tukey post hoc test (alpha=.05). RESULTS: Compared with the control group, Duralon and Harvard cements demonstrated antibacterial properties even after 3 months with the direct contact test (P <.002), while Ketac-Cem exhibited no antibacterial properties. In the agar diffusion test, no antibacterial activity was observed for any of the tested cements. The different powder/liquid ratios had a negligible effect on the antibacterial properties of the tested cements. CONCLUSIONS: Within the limitations of this study, Duralon and Harvard cements possessed prolonged antibacterial properties, while Ketac-Cem exhibited no antibacterial activity. The direct-contact test may be a more suitable test than the agar diffusion test to evaluate antibacterial properties of definitive cements. PMID- 15798689 TI - Surface roughness and fatigue performance of commercially pure titanium and Ti 6Al-4V alloy after different polishing protocols. AB - STATEMENT OF PROBLEM: Surface quality of cast metal is directly related to service performance under fatigue stress. Surface heterogeneities resulting from either finishing or polishing processes or by corrosive agents such as fluoridated solutions, can negatively affect fatigue life. Cast titanium frameworks are difficult to polish, and an accepted polishing protocol has not been established. PURPOSE: This study evaluated and compared surface roughness of cast commercially pure titanium (CP Ti) and Ti-6Al-4V alloy submitted to conventional or electrolytic polishing, correlating the results with corrosion fatigue strength testing performed in artificial fluoridated saliva. Specimens were also tested in air at room temperature to evaluate the effectiveness of the corrosion-fatigue test model. MATERIAL AND METHODS: For each metal, 40 dumb-bell shaped rods, 2.3 mm in diameter at the central segment, were cast. Conventional polishing was performed on 20 specimens of each metal following the manufacturer's instructions. A source of continuous electrical current was used for electrolytic polishing of the other 20 specimens of each metal, which were immersed in an electrolytic solution containing 5% fluoridric acid, 35% nitric acid, and 60% distilled water. Surface roughness, Ra (microm), was measured with a profilometer, and fatigue tests were carried out with a universal testing machine using a load 30% lower than the 0.2% offset yield strength. After failure, the fractured surfaces were examined using scanning electron microscopy. Surface roughness means were analyzed with a 2-way analysis of variance and the Tukey multiple comparisons test (alpha=.05). RESULTS: Electrolytic polishing (0.24 +/- 0.05 microm) provided significantly (P <.05) lower surface roughness values than conventional polishing (0.32 +/- 0.06 microm). Regardless of the polishing protocol, surface roughness of Ti-6Al-4V alloy (0.25 +/- 0.06 microm) was significantly lower (P <.05) than that of CP Ti (0.31 +/- 0.05 microm), and the fluoridated environment did not influence fatigue performance. There was no correlation between fatigue performance and surface roughness. CONCLUSION: Surface roughness of Ti-6Al-4V was significantly lower than that of CP Ti. For cast titanium frameworks, the electrolytic polishing regimen was found to be more effective than the manufacturer's polishing instructions with abrasives and rotary instruments. After polishing, differences in surface roughness values did not affect corrosion-fatigue performance. PMID- 15798690 TI - Restoring dentate appearance: a literature review for modern complete denture esthetics. AB - Despite the fact that solutions to functional and comfort problems are often available, successfully restoring the appearance of an edentulous patient remains a challenge. This review of the literature demonstrates the limited amount of useful evidence-based information available for restoration of dentate appearance in edentulous individuals. The English language peer-reviewed literature from 1880 to the present was reviewed. Articles were identified through previous literature reviews, an extensive hand search, and a search of MEDLINE using the key words esthetics and denture esthetics. Three main areas of information were found: published guidelines for achieving natural appearance, patient preference studies, and studies that have collected and analyzed anatomic norms. PMID- 15798691 TI - A technique for the fabrication of an immediate mandibular surgical stent securing a skin graft. AB - Patients who receive a skin graft following an ablative mandibular procedure may require an immediate intraoral surgical stent. This article describes an efficient and accurate method of fabricating an immediate mandibular surgical stent in the operating room setting. PMID- 15798692 TI - A double-impression technique for craniofacial defects. PMID- 15798693 TI - A technique of making impressions on patients with mandibular bony exotoses. PMID- 15798694 TI - Fabrication of removable partial denture remount casts prior to delivery. PMID- 15798695 TI - A time-saving technique for selection, insertion, and provisional restoration of angulated abutments for dental implants. PMID- 15798696 TI - [External counterpulsation: experience of institution of transplantology and artificial organs]. AB - External counterpulsation produces profound impact on hemodynamics especially in patients with myocardial ischemia. However the use of the method is not indicated in patients with left ventricular failure in whom in can provoke development of shock and pulmonary edema. PMID- 15798697 TI - [Effect of pacemaker implantation on clinical course of ischemic heart disease and choice of optimal pacing parameters]. AB - Effect of cardiac pacing on clinical course of ischemic heart disease was assessed in 154 patients with class II-IV angina pectoris with implanted pacemakers. Improvement of symptoms (decrease of number of anginal attacks, increase of exercise tolerance, reduction of number and doses of antianginal drugs) occurred in 72 patients (46.8%). In 30 patients (19.5%) increase of frequency of angina was accompanied with changed character, localization and duration of attacks as well as response to nitroglycerin. This was believed to be caused by augmented myocardial oxygen consumption due to 1.5-2 fold heart rate elevation during pacing and psychocardial syndrome. In 52 patients (33.8%) pacing was not associated with any changes of character of angina. It was shown that reprogramming of pacing parameters aimed at optimization of coronary reserve should be performed with consideration of angina class and presence of chronic heart failure. Optimal pacing rate was supposed to be 55-65 and 75-85 per min in patients with low coronary reserve and heart failure, respectively. PMID- 15798698 TI - [Lipid blood spectrum in patients with amiodarone associated hypothyroidism. Effect of replacement therapy with L-thyroxine]. AB - Blood lipid levels were measured in 23 patients with amiodarone associated hypothyroidism (most of them had ischemic heart disease). Abnormalities of lipid spectrum were found in 12 of these patients. All 12 patients were subjected to replacement therapy with l-thyroxine. Compensation of thyroid status was associated with average 12.6 and 12.3% lowering of total and low density lipoprotein cholesterol, respectively. However target levels of low density lipoprotein cholesterol were achieved only in 1 patient. There were no significant changes of high density lipoprotein cholesterol and triglycerides. PMID- 15798699 TI - [Level of endothelial vasodilatation and activity of angiotensin converting enzyme as criterion of efficacy of therapy in patients with mild and moderate hypertension]. PMID- 15798700 TI - [Recognition of actual periods of nocturnal rest from 24-hour heart rate profile]. AB - Inability to delineate exactly periods of nocturnal sleep and diurnal wakefulness during 24-hour blood pressure (BP) monitoring causes reporting of erroneous BP values for these periods. We suggested a simple mathematical algorithm for detection of periods of nocturnal rest determined as period of low values of cardiac rhythm using data of BP monitoring itself. AIM: To compare novel method of recognition of periods of sleep with 2 standard techniques: use of fixed time interval between 23 and 7 hours (1), or period of sleep according to patient's diaries (2). Reproducibility of nocturnal BP lowering between two 24-hour intervals during 48-hour blood pressure monitoring was used as a measure of precision of determination of diurnal/nocturnal BP. METHODS: Ambulatory 48-hour BP monitoring was carried out in 33 patients with uncomplicated stage II hypertensive disease. Automatic analysis of BP monitoring data was performed with the use of specially designed computer application. Standard deviation (SD) of differences (SDD) between pairs of nocturnal BP lowering during 48 hours was used as a measure of reproducibility. RESULTS: Reproducibility of values obtained with novel algorithm (SDD for systolic/diastolic BP 6.7/8.2 mm Hg) was substantially better than those obtained with standard methods (1) and (2) (SDD 13.0/14.8 and 13.5/18.3 mm Hg, respectively). CONCLUSION: The proposed method of recognition of the period of nocturnal rest substantially improved precision of automatic analysis of 24-hour BP monitoring. PMID- 15798701 TI - [Effect of low dose combination of antihypertensive drugs on the state of microcirculation]. PMID- 15798703 TI - [Chronic heart failure in patients with type-2 diabetes]. AB - Combination of diabetes and chronic heart failure is a frequently occurring syndrome. We studied interrelationship between state of carbohydrate metabolism, diabetic nephropathy and level of natriuretic peptide in patients with type 2 diabetes and concomitant chronic heart failure. The study confirmed high diagnostic value of natriuretic peptide as quantitative criterion of the presence of heart failure and revealed association of decompensation of carbohydrate metabolism and presence of diabetic nephropathy with more severe heart failure and left ventricular dysfunction. Among patients with renal failure those with more severe heart failure had low values of glycosylated hemoglobin. PMID- 15798702 TI - [Effect of long-term therapy with contemporary drugs on diastolic cardiac function in patients with chronic heart failure]. AB - Bisoprolol, carvedilol, enalapril are widely used for the treatment of patients with chronic heart failure. Definite role in the treatment of these patients is also played by angiotensin-II receptor blockers. Diastolic left ventricular function is widely spread among patients with chronic heart failure and its important value for prognosis has been demonstrated. However effect of modern drugs used for the treatment of chronic heart failure has been poorly studied. We assessed effects of enalapril, its combination with bisoprolol, carvedilol, and irbesartan on parameters of diastolic function in a randomized prospective controlled study on 84 patients with NYHA class III-IV heart failure and left ventricular ejection fraction <40%. It has been shown that left ventricular dysfunction is highly prevalent in patients with chronic heart failure and that differentiated approach to the choice of a drug for the treatment of chronic heart failure requires consideration of the type of diastolic dysfunction. PMID- 15798705 TI - [Potential of bone marrow mesenchymal stem cells for the treatment of diseases associated with damage of cardiac tissue]. AB - Ability of mononuclear and adhesive cells to form differentiated elements of various tissues and organs in tissue culture in vitro has been studied in experiments on CBA mice. It has been established that adhesion mesenchymal stem cells are present in bone marrow. These cells are able to generate muscular, nervous, endothelioid, epithelioid, reticular, fibroblastoid, chondrogenic, osteogenic, and other types of cells. Mesenchymal islets have been frequently found both as parts of colonies and as independent units. Potential developmental flexibility of mesenchymal cells allows to suppose their high efficacy in pathogenetic cell therapy of heart diseases when besides damaged muscular elements vascular and nervous elements should be also restored. Mononuclear cells have not demonstrated this ability. They formed mainly monocyte-macrophagal and lymphoid cells. PMID- 15798704 TI - [Protection of rat heart myocardium with a selective Na(+)/H(+) exchange inhibitor and ischemic preconditioning]. AB - Aim of this study was to compare effects of BIIB-722, a novel Na(+)/H(+) exchanger-1 inhibitor, and ischemic preconditioning (IP) on infarct size and metabolism of area at risk in rats. Regional ischemia was induced by 40-min occlusion of a diagonal branch of left anterior descending coronary artery (LAD); it was followed by 60-min reperfusion. Intravenous bolus injection of BIIB-722 (3 mg/kg) dissolved in 280 mM xylitol was performed before regional ischemia or during the first minute of reperfusion. In the control group 280 mM xylitol was infused before ischemia or at the beginning of reperfusion at the same mode. IP was initiated by two cycles of 5-min LAD occlusion followed by 5-min reperfusion prior to sustained regional ischemia. Microdialysis technique was used to monitor pH and inorganic phosphate (P(i)) in the interstitial fluid of the area at risk. Metabolic state of the area at risk was assessed by ATP, phosphocreatine (PCr) and lactate levels; cellular membrane damage was evaluated by total creatine (SigmaCr=PCr+Cr) tissue content. Myocardial infarct size was determined by computer planimetry after staining of left ventricular slices with 2,3,5 triphenyltetrazolium chloride. BIIB-722 administration before or after ischemia, as well as IP, had no effect on cardiac hemodynamics and acid-base indices of arterial blood throughout the experiments. The infarct size/area at risk ratio was 43.5+/-5.2% in control and was reduced to 11.4+/-3.1% with IP, and to 17.0+/ 3.6% and 25.8+/-2.6% with BIIB-722 infused on early reperfusion and before ischemia, respectively. BIIB-722 administration during the first minute of reperfusion as well as IP significantly augmented ATP and PCr contents, reduced lactate level and decreased ECr loss at the area at risk by the end of reperfusion as compared with values in control. Additionally significantly higher rates of pH recovery and reduction of P(i) concentration in the interstitial fluid were observed during reperfusion compared with these indices in control. BIIB-722 administration before ischemia had much effects on contents of energy and carbohydrate metabolites at area at risk. The results obtained indicate that ability of BIIB-722 to limit infarct size and improve metabolism in the area at risk is comparable to cardioprotective effects of IP. Therefore this study substantiates a possibility of application of a novel Na(+)/H(+) exchange inhibitor for clinical investigations. PMID- 15798706 TI - [Lamine gene mutations in dilated cardiomyopathy]. PMID- 15798708 TI - [Chronic pulmonary hypertension after acute pulmonary artery thromboembolism]. PMID- 15798707 TI - [Modification of lipid structure of cellular membrane in patients with ischemic heart disease with various extent of involvement of coronary vascular bed]. PMID- 15798710 TI - [Systemic thrombolysis: clinical functional assessment of electrical instability of the heart]. PMID- 15798709 TI - [Assessment of antihypertensive effect of telmizartan in patients with hypertensive disease]. PMID- 15798711 TI - [Endothelial dysfunction in cardiovascular diseases (Data from XIII European Conference on Arterial Hypertension)]. PMID- 15798717 TI - [Atrial fibrillation: choice of the method of pharmacological cardioversion]. AB - The paper considers circumstances under which it is expedient or not expedient to perform cardioversion in patients with paroxysmal or persistent atrial fibrillation. It contains discussion of benefits, limitations and drawbacks of electrical and pharmacological methods of cardioversion. American College of Cardiology/American Heart Association/European Society of Cardiology Guidelines for the Management of Patients With Atrial Fibrillation are presented. These guidelines suggest amiodarone, dofetilide, ibutilide, propafenone, flecainide, and quinidine as first line therapy because of their proven efficacy. Efficacy of intravenous disopyramide and procainamide in some patients with recent onset atrial fibrillation is recognized with certain reservations. The use of azimilide and dronedarone is considered promising. In Russia nibentan and ethacizine can be also used. Special emphasis is made on the possibility of wide use (including self-administration) of loading doses of oral propafenone for cardioversion in some categories of patients with atrial fibrillation of recent onset. PMID- 15798718 TI - [Technical approaches to registration of electrocardiogram]. PMID- 15798719 TI - [Scientific value of the EUROPA Study: mechanisms of beneficial effect of perindopril in atherosclerosis]. PMID- 15798720 TI - [Positron emission tomography with (18)F-fluoro-2-deoxyglucose in cardiological diagnosis]. AB - This clinically oriented review presents main principles of metabolism of cardiac muscle, pathophysiology of myocardial hibernation and stunning, as well as methodological principles of positron emission tomography (PET) of the heart with (18)F-fluoro-2-deoxyglucose ((18)F-FDG). Diagnostic and prognostic value of (18)F FDG PET and scintigraphic sings of disturbed myocardial viability, contractility and metabolism are also described. Efficacy of (18)F-FDG PET is compared with other imaging methods such as radionuclide, ultrasound and radiological. Literature data and clinical cases demonstrate importance of preoperative diagnosis of hibernating myocardium in patients with ischemic heart disease. (18)F-FDG PET is a basic method of detection of potentially reversible pathological states of the heart (hibernation and stunning); it has high sensitivity and specificity as well as predictive power in relation to forthcoming course of ischemic heart disease. This noninvasive method of investigation provides unique information on severity of ischemic heart disease for stratification of patients in risk groups and selection of candidates for coronary artery bypass surgery or cardiac transplantation. PMID- 15798721 TI - [Cytokines in complex treatment of posttraumatic empyema of the pleura]. AB - Results of treatment of 114 patients with posttraumatic empyema of the pleura (EP) after closed injury of the thorax (62), stab-incised (43) and missile (9) wounds are presented. In complex treatment of 25 patients one of cytokines - recombinant interleukin-1v (betaleukin) -- was administered intravenously (10 patients) and intrapleurally (15 patients). Control group consisted of 35 similar patients treated with traditional methods including drainage and sanitation of empyema's cavity with protheolytic enzymes, antibacterial and detoxication therapy. Comparative analysis has demonstrated that patients of the study group showed fast normalization of immunogram's parameters, decrease of duration of purulent process (51+/-4 days, on the average, in the control group and 28+/-3 days in the study group) due to fast obliteration of purulent cavity, and decrease of rate of chronic forms of EP (from 20 to 4%) which required surgical treatment. PMID- 15798722 TI - [Videoendosurgical treatment of postinjection abscesses and phlegmons]. AB - Method of treatment of postinjection abscesses and phlegmons was developed. This method includes endovideoscopy of purulent focus with creation of permanent liquid medium, visual revision of purulent cavity with evacuation of pus, controlled radical surgical treatment and introduction of perforated drainages for flowing drainage. Hand endoscopic instruments or special shaver were used for surgical necrectomy. Flowing-aspiration drainage of formed cavity was performed after surgery. This technique was used in 12 patients. It permitted to improve significantly results of treatment, reduce period of temporary disability and achieve good cosmetic effect. PMID- 15798724 TI - [Changes of motor-evacuatory function of the gastrointestinal tract after surgery in children with acute appendicitis]. AB - A new method of computed phonoenterography (CPEG) evaluates objectively motor evacuatory function of the gastrointestinal tract. Comparative study of changes in motor-evacuatory function of the intestine was carried out according to CPEG data in 60 children aged from 6 to 15 years after traditional surgery with Volkovich-Dyakonov incision, and after endovideosurgical appendectomy for plegmonous and gangrenous appendicitis. Inflammation of the appendix leads to inhibition of intestinal motor function. Low surgical trauma in endovideosurgical appendectomy promotes faster recovery of intestinal peristalsis according to CPEG. PMID- 15798723 TI - [Multispiral computed tomography in diagnosis of tumors of the mammary gland]. AB - For the first time, results of multispiral computed tomography (MSCT) with intravenous contrast enhancement in diagnosis of difficult case of breast tumors are presented. This method permits localization and staging of tumor. Contrast enhancement aids to reveal small tumors (less 1 cm). Thin sections and algorithm of high image resolution permit to diagnose microcalcinates which are early and important symptoms of malignant tumor. Evaluation of state of regional lymph nodes, analysis of bones permits to evaluate generalization of the process. MSCT mammography may be used as amplifying method in diagnosis of breast diseases. This method is also indicated in atypical location of cancer. Further studies of MSCT-mammography permit to increase diagnostic value of this method. PMID- 15798725 TI - [Surgical treatment of cancer of the stomach in patients over 70 years]. AB - Surgical treatment was performed in 322 patients over 70 years with cancer of the stomach. Radical operation was made in 220 (68.3%) patients: subtotal distal resection -- in 137, subtotal proximal resection -- in 18, gastrectomy -- in 55, extirpation of the gastric stump -- in 10. Symptomatic and test surgeries were performed in 102 (31.7%) patients. Postoperative complications were seen in 16,1+/-2,1% cases, lethality was 6.8+/-1.4%. Complications after radical resections were seen in 16.4+/-2.5% patients, lethality was 5.0+/-1.5%. The rate of purulent complications was maximal (8.4%). Cardiovascular complications and coagulopathies (4.7%) were the main causes of lethal outcome (in 11 of 15 patients). Score scale of assessment of postoperative risk was developed. Five years survival in stage I cancer of the stomach was 62.8%, stage II -- 42.6%, III -- 16.7%. Immediate and long-term results demonstrate necessity of radical surgical treatment of patients with cancer of the stomach over 70 years depending on the disease stage and risk factors. PMID- 15798726 TI - [Individual prognosis of severity of postoperative period and outcome of general peritonitis]. AB - Evaluation of severity according to SAPS scale and examination of dynamics of middle molecules in blood plasma (MCMn) were carried out in 167 patients with different surgical diseases complicated with general peritonitis. It is demonstrated that level of MCMn compared with SAPS scale data is objective index of endotoxicosis, severity of general peritonitis, and it can be regarded as criterion of prognosis of disease outcome. Dynamic assessment of MCMn level has the most diagnostic importance in groups of patients with 5 - 12 scores according to SAPS scale. Prognostic value of this index is small when infectious-toxic shock occurs. PMID- 15798727 TI - [Repeated surgeries after vagotomy in duodenal ulcers]. AB - Experience of surgical treatment of 73 patients with different postvagotomic syndromes was analyzed. The recurrent ulcers were seen most often -- in 54 (74.0%) patients. All the patients have undergone selective proximal vagotomy including isolated surgery in 19 patients and in combination with different drainage surgeries in 54 patients. High gastric secretion was the cause of recurrent ulcers in all the cases. Resection of the stomach was the most often reconstructive surgery -- 58 (79.5%) patients. Three (4.1%) patients died. It is concluded that postvagotomic syndromes are severe diseases of operated stomach with difficult treatment, further studies are necessary. PMID- 15798728 TI - [Surgical policy in echinococcosis of the liver with affection of bile ducts]. AB - Short-term results of surgical treatment of 277 patients with hepatic echinococcosis associated with affection of bile ducts are analyzed. If it is impossible, additional drainage of bile ducts and residual cavity is indicated. Pericystectomy is recommended when hydatid cyst has calcified fibrous capsule. It is demonstrated that rational surgical policy permits to achieve favorable results: postoperative complications were seen in 25.9% cases, lethality was 1.4%. PMID- 15798729 TI - [Treatment of epithelial coccygeal fistula]. AB - Analysis of treatment of 822 patients with epithelial coccygeal fistula has demonstrated that purulent complication is indication for primary radical surgery. Number of complications and recurrences decreases if the first focus of inflammation is treated and epithelial fistula is resected after healing of the wound. Anaerobic inflammation is seen in 0.4% patients with purulent epithelial coccygeal fistula. These patients require radical resection of affected tissues and general intensive therapy. There were no lethal outcomes after treatment of inflammatory complications. Hospital stay was 7.7 bed-days. All the surgical methods have disadvantages. Proposed method demonstrates the best results. Suppuration of the wounds was seen in 0.7% cases, recurrences -- in 1.1% patients. PMID- 15798730 TI - [Results of Shuldays' surgery in elective and urgent surgery]. AB - Results of treatment of inguinal hernias with multilayer plastic reconstruction (PR) according to Shuldays in elective and urgent surgery are analyzed. Advantages of multilayer PR are evaluated. In long-term period (up to 3 years) recurrences were seen in 0.5 to 1.0%. This is 10 times less than in plastic surgeries of anterior wall. Multilayer PR is particularly indicated for patients over 40 years with long anamnesis, weak tendinous structures, severe destruction of the posterior wall. This surgery is universal, can be performed in elective and urgent situations. PMID- 15798731 TI - [Storage of autologous hemocomponents in patients before endoprosthesis of great joints]. AB - Storage of autologous hemocomponents in patients prepared for elective surgery is the method of choice for adequate preparation before surgery. Autogenic erythrocytes and blood plasma increase infectious safety and efficacy of postoperative transfusion therapy. Up-to-date storage of autologous hemocomponents is indispensable condition for effective treatment of orthopedic patients. PMID- 15798732 TI - [Ultrasonic technologies in diagnosis and surgery planning in patients with nodular pathologies of the thyroid gland]. AB - Twenty patients of the control group without pathology of the thyroid gland were examined for development of the method of 3D-ultrasonic reconstruction and 3D angiography. The study group consisted of 74 patients with nodular pathologies of the thyroid gland, 53 of them underwent surgery. Ultrasonic data was compared with results of clinical and laboratory studies, puncture biopsy and intraoperative data. Three-dimensional ultrasonic pictures permitted to detect spatial orientation of the nodes relative to lobes and segments of the thyroid gland, vascular structures, to evaluate character of nodes vascularization, to analyze nature and invasiveness of the tumor. Based on these data optimal variant of surgery was chosen. In 12 cases virtual modeling of surgeries with 3D echography and ultrasonic angiography was performed. These methods permit to choose optimal variant of surgery and make possible prediction and prevention of complications. PMID- 15798733 TI - [Mini-invasive technologies in diagnosis and treatment of major vessels diseases]. AB - Experience with 150 endovascular stents and endoprostheses including ones of vertebral and carotid arteries (first experience in Russia) is analyzed. Mini invasive surgery is indicated for patients with isolated arterial lesions (segmental stenosis, short occlusions), changed and hard for surgery parts of vascular system (renal, visceral arteries, branches of aortic arch), restenosis after traditional surgeries and also for patients with severe concomitant diseases. Rate of complications of carotid endovascular stenting was 4,3%. The follow-up ranged from 2 months to 5,5 years. There were no restenoses in the stented zone. Complete absence of complains and regress of neurological symptoms were seen in 89% patients. PMID- 15798734 TI - [Choice of radical surgical method in patients with sutured perforated gastroduodenal ulcers]. AB - Long-term results of suturing of perforated gastroduodenal ulcer in 364 patients were analyzed. Within 16 years after surgery recurrence of ulcer was seen in 215 (62.9%) patients. Radical elective surgeries were performed in 197 (54.1%) from 2 mos to 16 years after perforation of ulcer. Indications for radical surgery were determined on the basis of the disease duration, presence and nature of complications, level of acid-production, etc., and also data of endoscopic ultrasonography. In early postoperative period 1 (0.5%) patient died. Long-term results were followed from 2 mos to 11 years in 168 (85.3%) patients. Recovery of reservoir and motor-evacuation function of the operated stomach was demonstrated with special examinations. The mean gastrointestinal index (GIQLI) in long-term period was 124.6 after resection surgeries and 120 after organ-retaining surgeries. PMID- 15798735 TI - [Evaluation of surgical traumaticity of standard and extended surgical procedures in cancer of a proximal part of the stomach]. AB - Analysis of the results and evaluation of surgical trauma of 454 various resections for cancer of a proximal part of the stomach were carried out. Proximal subtotal resection of the stomach with resection of the esophagus were performed in 151 patients (33.2% of all the resections). Gastrectomy was performed in 296 (65.2%) patients. Standard resection was carried out in 254 (55.9%) patients, extended -- in 72 (15.8%), combined -- in 64 (14,1%), extended combined -- in 64 (14,1%). It is demonstrated that number of complications after surgical treatment of cancer of a proximal part of the stomach is high enough due to traumaticity of abdominal-thoracic surgical procedures. Surgical approach, duration of surgery, intraoperative blood loss have significant influence on short-term results. Duration of surgery more than 300 min is the main factor making worse short-term results of extended resection of the stomach. In such duration of surgery postoperative complications were seen in 45.0+/-11.4%, lethal outcome -- in 30.0+/-10.5% cases. Extended surgical procedures demonstrate better long-term results. Five-years survival of patients with cancer of proximal part of the stomach (stage II and III) after extended surgeries is 53.3+/-13.3 and 33.3+/-6.3%, respectively. Standard resection is recommended for elderly patients with low functional reserves. PMID- 15798736 TI - [Complicated gastroduodenal ulcers]. AB - The results of the treatment of 2384 patients with gastric and duodenal ulcers are analyzed. It is demonstrated that over last years the number of complications has increased: perforations -- 2.6 fold, and bleedings -- 7 fold. Markers of a destructive injury of gastric and duodenal mucosa (helicobacter contamination, immunological disorders, level of tumor necrotic factor alpha, indices of catabolism of connective tissue) were studied. It is revealed that such disorders have direct influence on destruction of gastric and duodenal wall manifesting as life-threatening complications. PMID- 15798737 TI - [Policy and surgical treatment in malignant tumor of pancreatoduodenal zone]. AB - Experience in the treatment of 65 patients with tumors of pancreatoduodenal zone is analyzed, 36 of them underwent standard and radical pancreatoduadenal resections. Diagnostic value of preoperative examinations is analyzed, type of surgery is substantiated depending of localization and spread of tumor, degree of hepatodepression. Features of pancreatoduodenal resection are described in detail. Lethal outcome was in 1 case. It is concluded that radical surgeries can be more widely used in cancer of periampullar zone and chronic pseudotumoric pancreatitis. PMID- 15798738 TI - [Results of application of synthetic antioxidants in treatment of patients with destructive pancreatitis]. AB - Results of examination and treatment of 103 patients with pancreonecrosis are presented. Influence of antioxidant mexidol on acute pancreatitis (AP) was studied. It is demonstrated that mexidol in AP permits to slow down destructive processes in the pancreas, promotes localization of inflammation and normalization of clinical and laboratory picture of the disease that permits to decrease significantly number of severe complications of AP and reduce lethality. PMID- 15798739 TI - [Minimally invasive technologies in diagnosis and treatment of local purulent complications of destructive pancreatitis]. AB - Sixty patients with local purulent complications of destructive pancreatitis were treated. Interventional procedures under visual control were performed in 38 patients: in 26 (68.4%) -- under control of computed tomography and in 12 (31.6%) -- ultrasound-assisted procedures. Traditional surgeries were performed in 22 patients. Complete recovery was seen in 23 (60,6%) of 38 patients of the first group, all these patients have solitary, one-chamber and non-sequestered purulent cavities which are formed in 82.6% cases due to microfocal pancreonecrosis. Transcutaneous drainage was ineffective in 14 (36.8%) patients that required conversion to laparotomy. It is demonstrated that treatment policy in purulent necrotic complication of pancreonecrosis must be individual. Adequate method of local treatment must be determined by size and localization of purulent and necrotic lesions, presence of sequesters and septa. PMID- 15798740 TI - [Laparoscopy in early postoperative period in elderly and old patients]. AB - For diagnosis and treatment of early postoperative intraabdominal complications 118 laparoscopic procedures were performed in 95 patients aged 55 to 94 years. Examination was carried out from 2 hours to 17 days after the first surgery. Diagnostic laparoscopy after surgeries on the liver and bile ducts was performed in 51 patients (most often), after ones on the intestine -- in 12 patients, on the stomach and the duodenum -- in 7, on organs of the small pelvis -- in 6, after appendectomy -- in 6 and after herniotomy -- in 8 patients. Primary surgery was emergency and urgent in 76, elective -- in 19 patients. Results of treatment of similar complications were studied in 91 patients (control group) who underwent laparotomy without previous laparoscopy. Both groups of patients were comparable by sex, age, severity of disease and types of primary surgical procedures. It is demonstrated that diagnostic and curative laparoscopic procedures in early postoperative intraabdominal complications in elderly and old patients permit to reduce lethality 2.4 fold. PMID- 15798741 TI - [Balloon compression hemostasis in injuries of parenchymatous organs]. AB - Balloon compression method of hemostasis in injuries of parenchymatous organs is described. Developed device consists of inflatable balloon and double-lumen tube for drainage of abdominal cavity and filling of balloon with air. Hemostasis is achieved due to inflation of balloon and pressure on injured surface. Parameters of pressure and time of compression were determined in experimental study. This device can be used both in open and video-assisted laparoscopic surgeries. Described method was used in 29 patients (22 of them underwent laparoscopic surgery and in 7 patients the device was applied during laparotomy). This method permits to achieve reliable sutureless hemostasis, to control hemostasis and pressure on the wound, to drain abdominal cavity. PMID- 15798742 TI - [Xenogenous islets of Langerhans transplantation of insular cells without immunosuppressive therapy in the treatment of insulin-dependent diabetes]. AB - Results of experimental and clinical xenotransplantation without immunosuppressive therapy in the treatment of insulin-dependent diabetes are summarized. Morphologic study demonstrated possibility of long-term preservation of macroencapsular xenogeneous culture of the insular cells in the blood of the recipients without immunosuppressive therapy. Thirteen patients with diabetes underwent surgery according to the developed method. Laboratory and clinical examinations demonstrated an increasing level of C-peptide, immunoreactive insulin, a decrease of fructozamine and insulin demand to 70-85%. It is concluded that intravascular xenotransplantation is an effective procedure, and it may be regarded as alternative to allotransplantation. PMID- 15798743 TI - [Choice of plastic surgery in ventral hernias]. AB - Morphological changes in the zone of plastic surgery with polypropylene graft were studied in experiment. Results of treatment of 195 patients with ventral hernias were analyzed. Study group consisted of 86 (44.1%) patients who underwent hernioplasty with subaponeurotic localization of the graft. Subaponeurotic hernioplasty was performed in 109 (55.9%) patients (control group). Algorithm of choice of plastic surgery's method in ventral hernia was developed with consideration of localization and size of hernia that permitted to reduce rate of early postoperative complications and to improve long-term results. PMID- 15798744 TI - [Experimental-morphologic study of bone tissue reaction to carbon-containing material implantation with initiated X-ray contrast property]. AB - In experimental study on 15 rabbits (chinchilla) influence of titanium plates implanted lapped on adjacent tissues in the region of the lower jaw body (comparison group) and carbon material with added boron in the concentrations of 8 and 15% (the study group) was studied. Results of the experimental morphological investigation show that carbon-based materials with boron addition (with its content 8 and 15%) did not impede adaptive rebuilding of bone tissues and in particular bone structure regeneration in the process of reactive rebuilding of the "maternal" bone. Moreover, as the result of reactive processes developing in osseous tissues after implantation of the tested materials their successful integration in surrounding tissue structures was detected. PMID- 15798746 TI - [Factors of individual radiation sensitivity in caries prevalence in population of radiation polluted regions]. AB - Analysing dental diseases prevalence in population of radiation polluted regions after the Chernobyl accident, one should take into account not only the level of the soil contamination with radionuclides but also factors of individual sensitivity to radioactivity, main of which is the age of the exposed person. PMID- 15798745 TI - [Clinical aspects of use of preparations from seaweeds for caries and gingivitis prevention]. AB - Clinical application of seaweed's preparations has shown the appreciable preventive effect, which is connected, first of all, with structure of a water solution of a laminaria's mineral concentrate. This perspective direction contains the following ways of perfection of primary prevention of dental caries: improvement of salivary glands function, optimization of saturation of oral liquid by mineral components, amplification of autopurification of oral cavity, increase of dental tissue's resistance. Application of a fususe's water solution extract improves clinical condition of paradontium, reduces quantity of pathogenic microorganisms that testifies to high anti-inflammatory efficiency of an extract. PMID- 15798747 TI - [Oral fluid nitrate reductase complex activity in teenagers dependent on oral hygiene level and DMFT-index]. AB - This research is directed to determine the influence of carious process at the stage of forming of constant occlusion on nitratereductase complex activity in the oral liquid. The correlation between of nitratereductase complex activity and DMF index, as well as the sex and the age of children with variable and constant occlusions was discovered. It is possible to estimate objectively a condition of a oral cavity due to nitratereductase complex activity. PMID- 15798748 TI - [Teeth electroexcitability in periodontitis]. AB - Electroodontometric study of 756 intact teeth in 209 patients 18-65 years of age with normal periodontium and different stages of periodontal disease (periodontitis) showed that 96.8% of teeth in patients with mild periodontitis retained normal sensitivity, 68.4% -- with moderate periodontitis and 55% -- with severe periodontitis. Decreased pain threshold was detected in 3.2% of teeth of patients with mild periodontitis, 22.6% -- with moderate periodontitis and 12% -- with severe periodontitis. Increased pain threshold was not detected in teeth of patients with mild periodontitis, but in 9% with moderate periodontitis and in 29% with severe periodontitis it was obvious. PMID- 15798749 TI - [Gum hyperthrophy as side-effect of cyclosporine A therapy]. AB - The clinical and histopothologic findings in 48 patients with gingival fibrous hyperplasia (hyperthophy) associated with cyclosporin A therapy are described. PMID- 15798751 TI - [Supersorbicid in comprehensive treatment of acute pyoinflammatory processes in maxillofacial region]. AB - Antibiotic-containing Supersorbicid was used for drainage during treatment of 56 patients with abscesses and phlegmon of maxillo-facial region. Clinical laboratory and bacteriological assessment showed that in local application of antibiotic-containing Supersorbicid in the complex treatment of abscesses and phlegmon of maxillo-facial region liquidation of local and common signs of inflammation was faser and corbent accumulated microbes and their toxins and the wound cleaning from necrotic mass was faser that resulted in acceleration of wound healing by secondary adhesion. The results showed that leucocyte intoxication index was significantly decreased in the studied patients and the content of infectious agents was decreased too in patients in comparison with control subjects. PMID- 15798750 TI - [Integrated evaluation of chewing gums and candies influence in oral cavity]. AB - In a blind study on 210 medical students during 3 months the influence of 5 types of chewing gums with sugar substitutes (Orbit, Dirol, Stimorol) and 1 type of candy with sucrose upon saliva properties, saliva flow and oral microflora activity were examined. Flow rate of saliva, its pH (mixed saliva), influence upon test sugar and carbamide pH curves, concentrations of carbamide, Ca and phosphate in saliva were determined. It is shown than frequent and prolong use of chewing gums reduced stimulated and unstimulated saliva flow. Frequent use of chewing gums with carbamide provoked increased activity of ureapositive oral microflora. Degree of alkaline changes of mixed saliva pH in case of 3-month use of chewing gums Orbit and Stimorol pro Z was reduced. Chewing gums had weak hygienic efficacy. PMID- 15798752 TI - [A new method of sublingual salivary gland cyst treatment by silicone membrane]. AB - It is suggested to use silicone membrane in operations of plastic cystotomy in cases of ranula. 1-2-year follow-up results are good, no recurrence was observed. PMID- 15798753 TI - [Surgical treatment of cystous lesions of the jaws in children using biocomposite material]. AB - Assessment of surgical treatment of cystous lesions of the jaws in children was performed on the basis of the results of treatment of 25 patients with the use of granulated biocomposite material GAP-99. It was shown by the authors that this material allows one to shorten the time of bone wound healing, to improve cosmetic and functional effects during the hospital stage of treatment. In long term period full rehabilitation was achieved in patients operated with the use of this biocomposite material. PMID- 15798754 TI - [Long-term results and possible complications when subperiostal implants are used in patients with considerable jaw atrophy]. AB - Analysis of the results of subperiostal implants use in 130 patients with considerable atrophy of jaws bone tissues is presented. Early and delayed complications occurring in patients after subperiostal implants use are detailed. PMID- 15798755 TI - [Prevention of postoperative inflammatory complications in patients with maxillo facial region pathology]. AB - Results of the use of antibiotics for prevention and treatment of different maxillo-facial pathology in maxillo-facial surgery are presented. Antibiotics for prevention were employed in 185 patients and the results were compared with 165 patients serving as control group. For the prevention of postoperative complications single intravenous injections of antibiotics (Ceftriaxon, Rocifen, Lyncomycin) before narcosis were used in double doses. It was confirmed that this measure allow complete prevention of postoperative purulent complications in cases of closed mandibular fractures, reconstructive operations in maxillo-facial region, or reduce their frequency in cases of open fractures of mandible and nasal bones. PMID- 15798756 TI - [Follow-up analyses of surgical treatment of patients with deformities of nose end of congenital nature]. AB - Positive and negative features of the known techniques (suggested by Converse, Armstrong and Bourian) are discussed. The conclusion is made that it is necessary to modify the techniques to improve final results. PMID- 15798757 TI - [Results evaluation of surgical treatment of patients with lower eyelid defects]. AB - Surgical treatment of the patients with the lower lid defects are actual part of the maxillofacial surgeon's practice. Among different methods of treatment there are pedicle flaps, free skin flaps, tunnel flaps and hinged flaps, the latter are not widely used. However, as the study proved, hinded flaps have incontestable advantage and using it one can greatly widen their possibilities and improve plastic operations' RESULTS: The authors made an attempt to appreciate comparatively of the effectiveness of the various methods of the lower lid defects replacement. PMID- 15798758 TI - [Tooth bud fixation by the material LitAr]. AB - The results of the biotransformation investigation of the material LitAr after replacing through it the cystectomy area for the fixation of the permanent tooth rudiment are given here. The conversion process of the material LitAr into the bone tissue was checked roentgenologically and with the use of magnetic resonance image (MRI). In accordance with the MRI data all the bone tissue formation marks have been stated in 2 months after implanting the material LitAr into the rudiment fixation area. All the further observations (up to 10 months) have shown the bone tissue formation which was accompanied by cutting the tooth in the correct position and on time. PMID- 15798759 TI - [Correction of a vast craniofacial defect -- a case report]. AB - A case of a patient with a vast craniofacial defect after shooting is reported. The case is of interest because of a large size of the defect. The technique is described how to rehabilitate the patient with reconstruction the face contour and calvarium by silicone implants. PMID- 15798760 TI - Academic psychiatry in Brazil: confronting the challenges. PMID- 15798764 TI - Gastrin stabilises beta-catenin protein in mouse colorectal cancer cells. AB - As gastrin may play a role in the pathophysiology of gastrointestinal (GI) malignancies, the elucidation of the mechanisms governing gastrin-induced proliferation has recently gained considerable interest. Several studies have reported that a large percentage of colorectal tumours overexpress or stabilise the beta-catenin oncoprotein. We thus sought to determine whether gastrin might regulate beta-catenin expression in colorectal tumour cells. Amidated gastrin-17 (G-17), one of the major circulating forms of gastrin, not only enhanced beta catenin protein expression, but also one of its target genes, cyclin D1. Furthermore, activation of beta-catenin-dependent transcription by gastrin was confirmed by an increase in LEF-1 reporter activity, as well as enhanced cyclin D1 promoter activity. Finally, G-17 prolonged the tau(1/2) of beta-catenin protein, demonstrating that gastrin appears to exert its mitogenic effects on colorectal tumour cells, at least in part, by stabilising beta-catenin. PMID- 15798765 TI - Choice of geographic unit influences socioeconomic inequalities in breast cancer survival. AB - Socioeconomic differences in age-standardised crude survival for women diagnosed with breast cancer during 1991-1999 in England were influenced by the population of the geographic area used to assign the deprivation index, but not by the choice of index. PMID- 15798767 TI - Increased risk of thromboembolism in patients with malignant lymphoma: a single centre analysis. AB - An increased risk for thromboembolism in cancer patients has been observed in patients with solid tumours, whereas little data exist on malignant lymphoma. We found an overall thromboembolic event incidence of 7.7% in 1038 lymphoma patients treated in our institution, with a statistically significantly higher incidence in high-grade than in low-grade lymphoma. PMID- 15798766 TI - Second primary malignancies in patients with male breast cancer. AB - An international multicentre study of first and second primary neoplasms associated with male breast cancer was carried out by pooling data from 13 cancer registries. Among a total of 3409 men with primary breast cancer, 426 (12.5%) developed a second neoplasia; other than breast cancer, a 34% overall excess risk of second primary neoplasia, affecting the small intestine (standardised incidence ratio, 4.95, 95% confidence interval, 1.35-12.7), rectum (1.78, 1.20 2.54), pancreas (1.93, 1.14-3.05), skin (nonmelanoma, 1.65, 1.16-2.29), prostate (1.61, 1.34-1.93) and lymphohaematopoietic system (1.63, 1.12-2.29). A total of 225 male breast cancers was recorded after cancers other than breast cancer, but an increase was found only after lymphohaematopoietic neoplasms. BRCA2 (and to some extent BRCA1) mutations may explain the findings for pancreatic and prostate cancers. Increases at other sites may be related to unknown factors or to chance. This large study shows that the risks for second discordant tumours after male breast cancer pose only a moderate excess risk. PMID- 15798768 TI - Nasal-type NK/T cell lymphoma: clinical features and treatment outcome. AB - Nasal-type NK/T cell lymphoma is an increasingly recognised disease entity of aggressive clinical behaviour. The objective of this study was to investigate clinical features and treatment outcomes in patients with nasal-type NK/T cell lymphoma. From January 1991 to December 2003, 26 patients diagnosed as nasal-type NK/T cell lymphoma were included in the analysis. One half of patients presented with poor performance status (ECOG > or =2); 46% of patients were categorised as high intermediate or high-risk group according to IPI; and 46% of patients were diagnosed at advanced stage. The median survival for 26 patients with nasal-type NK/T cell lymphoma was 7.4 months (95% CI, 0.1, 16.9). The treatment outcome of primary anthracycline-based chemotherapy was poor: 60% CR rate in localised disease and 0% CR rate in advanced disease. After a median follow-up of 24.4 months (range 3.1-99.0) in patients with localised disease who had achieved a CR (range 29.6-165.7), three patients (50.0%) developed disease recurrence at 6.1, 21.8, and 52.1 months, respectively, and all patients presented with locoregional failure. The predictive factors for poor survival were of age greater than 60, advanced stage and poor performance in multivariate analysis. In conclusion, Nasal-type NK/T cell lymphomas showed a poor response to the conventional anthracycline-based chemotherapy, and thus an investigation for an innovative therapy is urgently needed to improve survival in these patients. PMID- 15798769 TI - Immune responses to the 105AD7 human anti-idiotypic vaccine after intensive chemotherapy, for osteosarcoma. AB - 105AD7 is a human monoclonal antibody that mimics the complement regulatory protein, CD55, overexpressed by many solid tumours including osteosarcoma. This study was designed to assess the toxicity and efficacy of this vaccine in a young age group of patients within 1-6 months of myleosuppressive chemotherapy. Out of 28, 20 (71%, 95% CI 51-87%) patients showed a significant T-cell proliferation response in vitro to the 105AD7 protein but not to human IgG. Furthermore, 13 out of 22 (59%, 95% CI 36-79%) patients showed antigen-specific gammaIFN secretion (range 20-370 U/ml). Nine out of 28 (32%, 95% CI 16-52%) patients made weak antibody responses to CD55. This study showed that 105AD7 was well tolerated in younger patients with osteosarcoma. In addition, two patients with possible clinical responses were given compassionate permission to continue immunisation quarterly for 2 years. They both remain alive and disease free 5.8 and 6.5 years from original diagnosis of osteosarcoma and showed no adverse effects of repeated immunisation. In conclusion, the majority of patients showed measurable T helper responses when vaccination was commenced within a 6-month window of intensive chemotherapy with no clinically significant toxicity. Future clinical trials incorporating immune stimulation strategies should include early introduction of vaccines during the highest risk period for relapse. PMID- 15798770 TI - Altered expression of topoisomerase IIalpha contributes to cross-resistant to etoposide K562/MX2 cell line by aberrant methylation. AB - KRN 8602 (MX2) is a novel morpholino anthracycline derivative having the chemical structure 3'-deamino-3'-morpholino-13-deoxo-10-hydroxycarminomycin hydrochloride. To investigate the mechanisms of resistance to MX2, we established an MX2 resistant phenotype (K562/MX2) of the human myelogeneous leukaemia cell line (K562/P), by continuously exposing a suspension culture to increasing concentrations of MX2. K562/MX2 cells were more resistant to MX2 than the parent cells, and also showed cross-resistance to etoposide and doxorubicin. Topoisomerase (Topo) IIalpha protein levels in K562/MX2 cells were lower of those in K562/P cells on immunoblot analysis and decreased expression of Topo IIalpha mRNA was seen in K562/MX2 cells. Topoisomerase II catalytic activity was also reduced in the nuclear extracts from K562/MX2 cells when compared with K562/P cells. Aberrant methylated CpG of Topo IIalpha gene was observed in K562/MX2 cells when compared with the parent line on methylation-specific restriction enzyme analysis. To overcome the drug resistance to MX2 and etoposide, we investigated treatment with 5-Aza-2'-deoxycytidine (5AZ), which is a demethylating agent, in K562/MX2 cells. 5-Aza-2'-deoxycytidine treatment increased Topo IIalpha mRNA expression in K562/MX2 cells, but not in K562/P cells, and increased the cytotoxicity of MX2 and etoposide. Methylated CpG was decreased in K562/MX2 cells after 5AZ treatment. We concluded that the mechanism of drug resistance to MX2 and etoposide in K562/MX2 cells might be the combination of decreased expression of Topo IIalpha gene and increased methylation, and that 5AZ could prove to be a novel treatment for etoposide resistant cell lines, such as K562/MX2. PMID- 15798771 TI - Ascites induces modulation of alpha6beta1 integrin and urokinase plasminogen activator receptor expression and associated functions in ovarian carcinoma. AB - Interactions between cancer cells and the surrounding medium are not fully understood. In this study, we demonstrate that ascites induces selective changes in the expression of integrins and urokinase plasminogen activator/urokinase plasminogen activator receptor (uPA/uPAR) in ovarian cancer cells. We hypothesise that this change of integrin and uPA/uPAR expression triggers signalling pathways responsible for modulating phenotype-dependent functional changes in ovarian cancer cells. Human ovarian surface epithelial (HOSE) cell lines and epithelial ovarian cancer cell lines were treated with ascites for 48 h. Ascites induced upregulation of alpha6 integrin, without any change in the expression of alphav, beta1 and beta4 integrin subunits. Out of the four ovarian cancer cell lines studied, ascites induced enhancement in the expression of uPA/uPAR in the more invasive OVCA 433 and HEY cell lines without any change in the noninvasive OVHS1 and moderately invasive PEO.36 cell lines. On the other hand, no change in the expression of alpha6 integrin or uPAR, in response to ascites, was observed in HOSE cells. In response to ascites, enhancement in proliferation and in adhesion was observed in all four ovarian cancer cell lines studied. In contrast, no significant increase in proliferation or adhesion by ascites was observed in HOSE cells. Ascites-induced expression of uPA/uPAR correlated with the increased invasiveness of HEY and OVCA 433 cell lines but was not seen in OVHS1, PEO.36 and HOSE cell lines. Upregulation of alpha6 integrin and uPA/uPAR correlated with the activation of Ras and downstream Erk pathways. Ascites-induced activation of Ras and downstream Erk can be inhibited by using inhibitory antibodies against alpha6 and beta1 integrin and uPAR, consistent with the inhibition of proliferation, adhesion and invasive functions of ovarian cancer cell lines. Based on these findings, we conclude that ascites can induce selective upregulation of integrin and uPA/uPAR in ovarian cancer cells and these changes may modulate the functions of ovarian carcinomas. PMID- 15798772 TI - Effective treatment of anal cancer in the elderly with low-dose chemoradiotherapy. AB - Chemoradiotherapy (CRT) is accepted as the standard initial treatment for squamous cell anal cancer. However, frail elderly patients cannot always tolerate full-dose CRT. This paper reports the results of a modified regimen for this group of patients. In all, 16 patients with biopsy-proven squamous cell carcinoma of the anal canal or margin and performance status or co-morbidity precluding the use of full-dose CRT were included in this protocol. The median age was 81 (range 77-91). Patients received a dose of 30 Gy to the gross tumour volume plus 3 cm margin in all directions. Concurrent chemotherapy comprised 5-fluorouracil 600 mg m(-2) given over 24 h on days 1-4 of radiotherapy. The treatment was well tolerated. All 16 patients completed treatment as planned. Only one patient experienced any grade 3 toxicity (skin). The local control at a median follow-up of 16 months was 73% (13 out of 16). The overall survival was 69% and disease specific survival 86%. This is a well-tolerated regimen for elderly/poor performance patients with anal cancer, which can achieve high rates of local control and survival. Longer follow-up will determine whether these encouraging results are maintained. PMID- 15798773 TI - Epigenetic alteration at the DLK1-GTL2 imprinted domain in human neoplasia: analysis of neuroblastoma, phaeochromocytoma and Wilms' tumour. AB - Epigenetic alterations in the 11p15.5 imprinted gene cluster are frequent in human cancers and are associated with disordered imprinting of insulin-like growth factor (IGF)2 and H19. Recently, an imprinted gene cluster at 14q32 has been defined and includes two closely linked but reciprocally imprinted genes, DLK1 and GTL2, that have similarities to IGF2 and H19, respectively. Both GTL2 and H19 are maternally expressed RNAs with no protein product and display paternal allele promoter region methylation, and DLK1 and IGF2 are both paternally expressed. To determine whether methylation alterations within the 14q32 imprinted domain occur in human tumorigenesis, we investigated the status of the GTL2 promoter differentially methylated region (DMR) in 20 neuroblastoma tumours, 20 phaeochromocytomas and, 40 Wilms' tumours. Hypermethylation of the GTL2 promoter DMR was detected in 25% of neuroblastomas, 10% of phaeochromocytoma and 2.5% of Wilms' tumours. Tumours with GTL2 promoter DMR hypermethylation also demonstrated hypermethylation at an upstream intergenic DMR thought to represent a germline imprinting control element. Analysis of neuroblastoma cell lines revealed that GTL2 DMR hypermethylation was associated with transcriptional repression of GTL2. These epigenetic findings are similar to those reported in Wilms' tumours in which H19 repression and DMR hypermethylation is associated with loss of imprinting (LOI, biallelic expression) of IGF2. However, a neuroblastoma cell line with hypermethylation of the GTL2 promoter and intergenic DMR did not show LOI of DLK1 and although treatment with a demethylating agent restored GTL2 expression and reduced DLK1 expression. As described for IGF2/H19, epigenetic changes at DLK1/GTL2 occur in human cancers. However, these changes are not associated with DLK1 LOI highlighting differences in the imprinting control mechanisms operating in the IGF2-H19 and DLK1-GTL2 domains. GTL2 promoter and intergenic DMR hypermethylation is associated with the loss of GTL2 expression and this may contribute to tumorigenesis in a subset of human cancers. PMID- 15798774 TI - A thromboxane effect of a hydroxytyrosol-rich olive oil wastewater extract in patients with uncomplicated type I diabetes. AB - OBJECTIVE: To assess the antioxidant/non-antioxidant effects of a hydroxytyrosol (HT)-rich phenolic extract from olive mill wastewaters administered with a breakfast. DESIGN, SETTING AND SUBJECTS: Five type I diabetic patients received 25 mg of HT the first day and 12.5 mg/day the following 3 days. Blood sampling was carried out at T(0) (baseline) and T(4d) just before the breakfast + HT administration and at time points 1, 2, 3 and 4 h after T(0). Urines (24-h) were collected from T(0) to T(4d). Baseline HbA1c was generally inferior to 10%, glycemia was within the range 6-24 mmol/l, whereas total cholesterol, HDL-chol and triglycerides were normal. RESULTS: The major finding was the 46% decrease in the serum TXB(2) production after blood clotting at T(4d). Plasma vitamin A, E, beta-carotene were not changed. Vitamin C tended to increase (P = 0.075). Plasma antioxidant capacity was enhanced at T(0)+1 h only, whereas its main determinants (albumin, bilirubin, uric acid) were not modified. Urinary 8-isoPGF(2alpha) levels were highly variable and were not affected significantly by HT administration. CONCLUSION: The major effect of HT accounts for an antiaggregating platelet action, leading to a possible prevention of thrombotic and microthrombotic processes. PMID- 15798775 TI - Prediction of DXA-determined whole body fat from skinfolds: importance of including skinfolds from the thigh and calf in young, healthy men and women. AB - OBJECTIVE: To investigate the relationship of percent body fat (%fat), assessed by dual energy-X-ray absorptiometry (DXA) or a four-compartment model, with upper body and lower limb skinfolds. DESIGN: Cross-sectional design involving forward stepwise and hierarchical multiple regression analyses to assess the relationship of %fat with skinfolds and a combination of four commonly used upper body skinfolds (biceps, triceps, subscapular and iliac crest) with the calf and thigh skinfolds. SETTING: University research laboratory. SUBJECTS: In all, 31 females, mean age 20.9 (+/-2.0) y, and 21 males, mean age 22.3 (+/-5.5) y volunteered for this study, which was approved by the Ethics Committee of the School of Sport, Health and Exercise Sciences, University of Wales, Bangor. MEASUREMENTS: %fat from DXA in both groups, and %fat from a four-compartment (water, bone mineral mass, fat and residual) model (%fat4C) in females only. Skinfolds were measured at the abdomen, iliac crest, biceps, triceps, subscapular, calf and thigh. RESULTS: All skinfolds were positively associated with DXA estimates of %fat (P < 0.01). In males and females, the thigh skinfold had the highest correlation with %fat. This was also observed when %fat4C was used as the criterion in females. Stepwise multiple regression analysis using %fatDXA as the criterion selected the thigh (R(2) = 0.82), calf (R(2) change 0.04) and iliac crest (R(2) change = 0.03) for females, and the thigh (R(2) = 0.79), iliac crest (R(2) change = 0.11) and abdomen (R(2) change = 0.03) for males (all P < 0.01). When %fat4C was used as the criterion in the females, only the thigh was selected as a significant predictor (R(2) = 0.76). Independent prediction factors were created from the sum of biceps, triceps, subscapular and iliac crest (sigma4skf) and from the sum of the thigh and calf (sigmathigh + calf). These factors were then entered into a hierarchical multiple linear regression analysis to predict percent fat. Order of entry was varied to allow the assessment of unique variance accounted for by each predictor. The sum of the thigh and calf explained more variance in %fatDXA than that explained by the sigma4skf alone, irrespective of the order of entry in both males and females. This was also observed when %fat4C was used as the criterion in the females. CONCLUSIONS: The results of this study confirm that lower body skinfolds are highly related to percent body fat in fit and healthy young men and women, and uphold current recommendations by the British Olympic Association to include the thigh skinfold with sigma4skf. Conventional use of the sigma4skf to estimate percent body fat is significantly enhanced by the inclusion of the thigh and calf skinfolds, either independently or in combination. In this group of males and females, the sum of the thigh and calf skinfolds accounted for the most variance in percent fat. PMID- 15798776 TI - Validity and reproducibility of resting metabolic rate measurements in rural Bangladeshi women: comparison of measurements obtained by Medgem and by Deltatrac device. AB - OBJECTIVE: To assess reproducibility and validity of resting metabolic rate (RMR) of Bangladeshi women as measured with the MedGem device and using the Deltatrac metabolic monitor as a reference; and (2) to evaluate the FAO/WHO/UNU basal metabolic rate (BMR)-prediction equations. DESIGN: In each of two sessions, resting oxygen consumption was measured in triplicate by MedGem and in triplicate by Deltatrac device. SETTING: Matlab area, the rural field research area of the Centre for Health and Population Research, Bangladesh (ICDDR,B). SUBJECTS: A total of 37 nonpregnant, nonlactating women, aged 27.6 +/- 4.5 y, BMI 20.8 +/- 3.1 kg/m(2) participated. RESULTS: The difference in oxygen consumption by MedGem and Deltatrac device was significantly level dependent. Within-subject within session variations (expressed as CV) were 9.0 and 3.0% (P < 0.01) and within subject between-session variations were 8.2 and 4.5% (P < 0.01) for MedGem and Deltatrac, respectively. Mean RMR measured by Deltatrac (5.17 +/- 0.51 MJ/day) was not significantly different from the BMR predicted by the FAO/WHO/UNU equations (5.16 +/- 0.42 MJ/day) in the second session and only 0.19 MJ/day higher than predicted in the first session (P < 0.05). CONCLUSION: Reproducibility and validity of the MedGem device was poor compared to the Deltatrac reference method. The FAO/WHO/UNU BMR-prediction equations give a good estimation of the BMR of rural, nonpregnant, nonlactating Bangladeshi women of 18 35 y. SPONSORSHIP: Wageningen University (The Netherlands) and ICDDR,B (Bangladesh). PMID- 15798777 TI - Mutational analysis of the TSC1 and TSC2 genes in a diagnostic setting: genotype- phenotype correlations and comparison of diagnostic DNA techniques in Tuberous Sclerosis Complex. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder characterised by the development of hamartomas in multiple organs and tissues. TSC is caused by mutations in either the TSC1 or TSC2 gene. We searched for mutations in both genes in a cohort of 490 patients diagnosed with or suspected of having TSC using a combination of denaturing gradient gel electrophoresis, single-strand conformational polymorphism, direct sequencing, fluorescent in situ hybridisation and Southern blotting. We identified pathogenic mutations in 362 patients, a mutation detection rate of 74%. Of these 362 patients, 276 had a definite clinical diagnosis of TSC and in these patients 235 mutations were identified, a mutation detection rate of 85%. The ratio of TSC2:TSC1 mutations was 3.4:1. In our cohort, both TSC1 mutations and mutations in familial TSC2 cases were associated with phenotypes less severe than de novo TSC2 mutations. Interestingly, consistent with other studies, the phenotypes of the patients in which no mutation was identified were, overall, less severe than those of patients with either a known TSC1 or TSC2 mutation. PMID- 15798778 TI - Molecular evolution: lateral gene transfer and other possibilities. PMID- 15798779 TI - Social interaction deficits caused by chronic phencyclidine administration are reversed by oxytocin. AB - Chronic administration of phencyclidine (PCP) has been advanced as a valid animal model of the social deficit symptoms of schizophrenia. In these studies, the cumulative time that male rats treated once a day for 14 days with PCP actively engaged in social behavior was decreased approximately 75% relative to saline treated control animals. In addition, these socially impaired rats had an increase in the relative amount of noncontact interactions compared with saline injected peers. Social behaviors were preferentially affected by PCP treatment because in two anxiety-related behavioral assays, the open field and light/dark emergence tests, there was a failure to differentiate between the PCP-treated rats and saline-injected control rats. Considering the general importance of the neuropeptides oxytocin and vasopressin in male social behaviors, studies of molecular markers related to these neuropeptides were performed. Hypothalamic oxytocin mRNA expression was significantly decreased while oxytocin receptor binding was increased in the central nucleus of the amygdala following chronic PCP treatment. Given the significance of central nucleus of the amygdala in social behavior, oxytocin was infused into the central nucleus of experimental and control male rats, and their postinfusion social interaction and open field behaviors were analyzed. A bilateral infusion of 1 mug of oxytocin into the central amygdala selectively restored the normal quantity and quality of social behavior in chronic PCP-treated male rats without altering open field behaviors. These findings suggest that deficits in the central oxytocinergic system may underlie the social impairment exhibited in this animal model of schizophrenia. PMID- 15798780 TI - ERbeta-selective estrogen receptor modulators produce antianxiety behavior when administered systemically to ovariectomized rats. AB - 17beta-Estradiol (E2) may influence anxiety behavior; however, its effects and mechanisms are not well understood. To determine whether E2's effects on anxiety behavior may involve actions at intracellular estrogen receptor (ER) alpha or beta isoforms, selective ER modulators (SERMs) were administered (10 microg; s.c.) to ovariectomized rats 48 h before testing for anxiety behavior. Rats received sesame oil vehicle, 17beta-E2, which has a high affinity for ERalpha and ERbeta, or SERMs that vary in their activity at ERalpha and beta. ERalpha selective SERMs were propyl pyrazole triol (PPT), which has more selective effects at ERalpha, than does the other ERalpha SERM utilized, 17alpha-E2, which also binds ERbeta. ERbeta-selective SERMs were diarylpropionitrile (DPN) and 7,12 dihydrocoumestan (coumestrol). DPN is more selective at ERbeta than coumestrol, which also binds ERalpha. 17beta-E2 and ERbeta-selective SERMs (DPN, coumestrol) produced clear antianxiety behavior in the open field, elevated plus maze, emergence, light-dark transition, defensive freezing, and Vogel punished drinking tasks. Anxiety behavior of rats administered ERalpha-selective SERMs (PPT, 17alpha-E2) was not different from vehicle; however, PPT and 17alpha-E2 enhanced sexual receptivity in a manner similar to 17beta-E2. Coadministration of tamoxifen (10 mg/kg) blocked the antianxiety behavior produced by 17beta-E2, DPN, or coumestrol. Together, these data suggest that actions at ERbeta may underlie some of E2's antianxiety effects. PMID- 15798781 TI - Rats maintained chronically on buprenorphine show reduced heroin and cocaine seeking in tests of extinction and drug-induced reinstatement. AB - Buprenorphine is being introduced as a maintenance therapy in opioid addiction, but it is not clear how buprenorphine will affect co-use of cocaine in opioid users. We examined the effects of chronic buprenorphine (BUP0: 0.0 mg/kg/day; BUP1.5: 1.5 mg/kg/day; BUP3: 3.0 mg/kg/day) on the locomotor activity effects of acute heroin (0.25 mg/kg, subcutaneously (s.c.)) and cocaine (20 mg/kg, intraperitoneally (i.p.)). Buprenorphine had no effect on the stimulatory effect of heroin, but potentiated the locomotor response to cocaine. To investigate further the interactions between buprenorphine (BUP1.5 and BUP3), heroin (0.125, 0.25 and 0.375 mg/kg, s.c.), and cocaine (10, 20 and 30 mg/kg, i.p.), we used in vivo microdialysis and high-performance liquid chromatography to analyze extracellular levels of dopamine (DA) in the nucleus accumbens (NAc). Buprenorphine attenuated the heroin-induced rise in NAc DA, but greatly potentiated the cocaine-induced rise. Finally, we examined the potential of the highest dose of buprenorphine (BUP3) to reduce heroin and cocaine seeking in the presence of drug-associated cues under extinction conditions and in tests for reinstatement induced by heroin (0.25 mg/kg, s.c.), cocaine (20 mg/kg, i.p.), and 15-min footshock stress (0.8 mA, 0.5 s/shock, 40 s mean OFF time) in rats trained to self-administer both drugs. Buprenorphine reduced heroin and cocaine seeking during extinction and following acute heroin and cocaine priming injections, but had no effect on stress-induced reinstatement. These results indicate that the suppression of responding following priming injections of drugs did not result from reduced motor activity, but possibly from a reduction in the salience of drug-associated cues induced by chronic buprenorphine treatment. PMID- 15798782 TI - Repetitive transcranial magnetic stimulation for the treatment of depression: to stimulate or not to stimulate? PMID- 15798783 TI - Efficacy of rapid-rate repetitive transcranial magnetic stimulation in the treatment of depression: a systematic review and meta-analysis. AB - OBJECTIVE: To systematically review the literature pertaining to rapid-rate repetitive transcranial magnetic stimulation (rTMS) compared with sham therapy for the treatment of a major depressive episode in order to arrive at qualitative and quantitative conclusions about the efficacy of rapid-rate rTMS. METHODS: MEDLINE, the Cochrane Library, the metaRegister of Controlled Trials and abstracts from scientific meetings were searched for the years 1966 until July 2003. The search terms "transcranial magnetic stimulation" and "transcranial magnetic stimulation AND depression" were used. Eighty-seven randomized controlled trials investigating the efficacy of rTMS were referenced on MEDLINE. Nineteen of these involved treatment of a major depressive episode, and these were reviewed. Six met more specific inclusion criteria including the use of rapid-rate stimulation, application to the left dorsolateral prefrontal cortex, evaluation with the 21-item Hamilton Rating Scale for Depression (HAM-D) and use of an intent-to-treat analysis. Scores on the 21-item HAM-D after treatment and standard deviations were extracted from each article for treatment and control subjects. A random-effects model was chosen for the meta-analysis, and the weighted mean difference was used as a summary measure. RESULTS: Six studies that met the inclusion criteria were identified and included in the meta-analysis. Two of these reported a significantly greater improvement in mood symptoms in the treatment versus the sham group. When combined in the meta-analysis, the overall weighted mean difference was -1.1 (95% confidence interval -4.5 to 2.3), and the results of a test for heterogeneity were not significant (chi2(5) = 5.81, p = 0.33). CONCLUSIONS: This meta-analysis suggests that rapid-rate rTMS is no different from sham treatment in major depression; however, the power within these studies to detect a difference was generally low. Randomized controlled trials with sufficient power to detect a clinically meaningful difference are required. PMID- 15798784 TI - A framework for targeting alternative brain regions with repetitive transcranial magnetic stimulation in the treatment of depression. AB - It has been argued that clinical depression is accompanied by reductions in cortical excitability of the left prefrontal cortex (PFC). In support of this, repetitive transcranial magnetic stimulation (rTMS), which is a method of enhancing cortical excitability, has shown antidepressant efficacy when applied over the left PFC, although the overall therapeutic effects remain inconclusive. The cerebral pathophysiology of depression is, however, not limited to dysfunctions in the PFC, thus, targeting alternative brain regions with rTMS may provide new therapeutic windows in the treatment of depression. Evidence from electroencephalography and lesion studies suggests that not only is the left PFC involved in depression but also the parietal cortex and cerebellum. Furthermore, rTMS over the parietal cortex and the cerebellum has been found to improve mood and emotional functioning, at least in healthy volunteers. We have integrated these findings in an rTMS-oriented theoretical framework for the neurobiology of low mood and depression. To establish the possible therapeutic efficacy of this model, whereby, for example, the application of slow rTMS over the right parietal cortex and fast rTMS over the cerebellum may be beneficial in different subtypes of depression, clinical rTMS studies that target the parietal cortex and cerebellum are warranted. PMID- 15798785 TI - Methylphenidate hydrochloride improves cognitive function in patients with advanced cancer and hypoactive delirium: a prospective clinical study. AB - OBJECTIVE: To investigate the clinical improvement observed in patients with advanced cancer and hypoactive delirium after the administration of methylphenidate hydrochloride. METHODS: Fourteen patients with advanced cancer and hypoactive delirium were seen between March 1999 and August 2000 at the Palliative Care Day Hospital and the inpatient Tertiary Palliative Care Unit of Montreal General Hospital, Montreal. They were chosen for inclusion in a prospective clinical study on the basis of (1) cognitive failure documented by the Mini-Mental State Examination (MMSE), (2) sleep-wake pattern disturbances, (3) psychomotor retardation, (4) absence of delusions or hallucinations, and (5) absence of an underlying cause to explain the delirium. All patients were treated with methylphenidate, and changes in their cognitive function were measured using the MMSE. RESULTS: All 14 patients showed improvement in their cognitive function as documented by the MMSE. The median pretreatment MMSE score (maximum score 30) was 21 (mean 20.9, standard deviation [SD] 4.9), which improved to a median of 27 (mean 24.9, SD 4.7) after the first dose of methylphenidate (p < 0.001, matched, paired Wilcoxon signed rank test). One patient died before reaching a stable dose of methylphenidate. In the other 13 patients, the median MMSE score further improved to 28 (mean 27.8, SD 2.4) (p = 0.02 compared with the median MMSE score documented 1 hour after the first dose of methylphenidate). All patients showed an improvement in psychomotor activities. CONCLUSIONS: Hypoactive delirium that cannot be explained by an underlying cause (metabolic or drug-induced) in patients with advanced cancer appears to be a specific syndrome that could be improved by the administration of methylphenidate. PMID- 15798786 TI - Changes in 5-HT1A receptor binding and G-protein activation in the rat brain after estrogen treatment: comparison with tamoxifen and raloxifene. AB - OBJECTIVE: It is thought that an imbalance in serotonergic neurotransmission may underlie many affective disorders. Thus, the serotonin-1A (5-HT1A) receptor is a target for antidepressant and neuroleptic drugs. It has been reported that estrogens modulate serotonergic neurotransmission. Therefore, we investigated the effect of long-term ovariectomy on 5-HT1A receptor-specific binding and G-protein activation in the brain. Correction therapy with estradiol was compared with treatments using the selective estrogen receptor modulators tamoxifen and raloxifene. METHODS: Four months after ovariectomy, Sprague-Dawley rats were treated with vehicle, 17beta-estradiol (80 microg/kg), tamoxifen (1 mg/kg) or raloxifene (1 mg/kg) subcutaneously for 2 weeks. Specific binding to 5-HT1A receptors was assessed by autoradiography of brain sections using the 5-HT1A agonist [3H]8-OH-DPAT. 5-HT1A receptor stimulation was measured using R-(+)-8-OH DPAT-stimulated [35S]GTPgammaS-binding autoradiography. RESULTS: Ovariectomy decreased uterine weight, which was corrected by estradiol; tamoxifen and raloxifene partially corrected this decrease. Hormonal withdrawal and replacement left [3H]8-OH-DPAT-specific binding unchanged in the cortex. In contrast, ovariectomy induced a decrease in R-(+)-8-OH-DPAT-stimulated [35S]GTPgammaS specific binding in the cortex; this was corrected by estradiol but was not corrected significantly by tamoxifen or raloxifene. In the hippocampus, ovariectomy had no effect on [3H]8-OH-DPAT-specific binding, whereas only 17beta estradiol treatment decreased this binding in a subregion of the CA3. Ovariectomy increased R-(+)-8-OH-DPAT-stimulated [35S]GTPgammaS-specific binding in the dentate gyrus (but not in the CA1 or CA3); this was corrected by estradiol and raloxifene, but not by tamoxifen. In the dorsal raphe nucleus, ovariectomy increased [3H]8-OH-DPAT-specific binding and R-(+)-8-OH-DPAT-stimulated [35S]GTPgammaS-specific binding; estradiol corrected this increase, but this was not corrected significantly by tamoxifen or raloxifene. CONCLUSIONS: An overall stimulation by estradiol of 5-HT1A receptor-specific binding and coupling was observed, decreasing raphe somatodendritic receptors and increasing cortical postsynaptic receptors. PMID- 15798787 TI - Perinatal complications in children with attention-deficit hyperactivity disorder and their unaffected siblings. AB - OBJECTIVES: Genetic and nonshared environmental factors (experienced by 1 family member to the exclusion of the others) have been strongly implicated in the causes of attention-deficit hyperactivity disorder (ADHD). Pregnancy, labour/delivery and neonatal complications (PLDNC) have often been associated with ADHD; however, no investigations aimed at delineating the shared or nonshared nature of these factors have been reported. We aimed to identify those elements of the PLDNC that are more likely to be of a nonshared nature. METHODS: We used an intrafamily study design, comparing the history of PLDNC between children diagnosed with ADHD, according to the criteria of the Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV), and their unaffected siblings. Children with ADHD were recruited from the outpatient, day treatment program of the Child Psychiatry Department, Douglas Hospital, Montreal. The unaffected sibling closest in age to the child with ADHD was used as a control. The history of PLDNC was assessed using the Kinney Medical and Gynecological Questionnaire and the McNeil-Sjostrom Scale for both children with ADHD and their siblings. Seventy children with ADHD along with 50 of their unaffected siblings agreed to participate in the study. Child Behavior Checklist (CBCL), Continuous Performance Test (CPT) and Restricted Academic Situation Scale (RASS) scores were also used as measures of ADHD symptoms in children with ADHD. RESULTS: The children with ADHD had significantly higher rates of neonatal complications compared with their unaffected siblings (F4,196 = 3.67, p < 0.006). Furthermore, neonatal complications in the children with ADHD were associated with worse CBCL total and externalizing scores and with poorer performance on the CPT. CONCLUSIONS: These results suggest that neonatal complications are probably a nonshared environmental risk factor that may be pathogenic in children with ADHD. PMID- 15798789 TI - Autistic-like findings associated with a urea cycle disorder in a 4-year-old girl. AB - A 4-year-old girl presented at our clinic with autistic-like symptoms, aggressivity and occasional hyperactivity. She had no history of neurologic or physical symptoms. Her condition was diagnosed as pervasive developmental disorder not otherwise specified, according to the criteria of the Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV). She received pharmacologic (thioridazine), educational and speech therapy. During this process, a urea cycle disorder was also identified, namely, ornithine transcarbamylase deficiency and arginase deficiency, because of the high level of ammonia in the patient's bloodstream, the high level of organic acids in the 24 hour urine collection and the constant presence of slow multifocal epileptic discharges on the electroencephalograms. The patient's protein intake was restricted, and she was treated with sodium benzoate and arginine. After 1 year of treatment, the autistic-like findings and hyperactivity were no longer apparent. PMID- 15798788 TI - Cortical hyperexcitability in post-traumatic stress disorder secondary to minor accidental head trauma: a neurophysiologic study. AB - OBJECTIVE: We applied paired transcranial magnetic stimulation (pTMS) to patients with post-traumatic stress disorder (PTSD) secondary to minor accidental head trauma. Our purpose was to determine the potential abnormality of motor cortex excitability in this pathologic condition. METHODS: pTMS stimulation, according to the conditioning-test paradigm employing interstimulus intervals (ISIs) of 1-6 ms, was used to investigate intracortical inhibition in control subjects and patients with PTSD. The study population consisted of 14 patients who had developed PTSD following minor head trauma, 12 healthy volunteers without a clinical history of head trauma and 11 healthy subjects who had reported accidental minor head trauma 1-4 months before the study. This clinical electrophysiologic study was performed at the Department of Neuroscience, University of Rome "Tor Vergata." RESULTS: All patients with PTSD exhibited a significantly lower motor evoked potential (MEP) inhibition than controls at 2 ms, 3 ms and 4 ms ISI. The statistical analysis of the pTMS protocol showed a significant effect (F2,36 = 25.63, p < 0.001) of the factor "group," because patients with PTSD showed a mean conditioned MEP amplitude higher than that observed in both control groups for all 6 ISIs analyzed. The "ISI" factor was also significant (F5,180 = 89.85, Greenhouse-Geisser epsilon = 0.35; p < 0.001), with the mean conditioned MEP amplitude increasing from 22.5% to 127.8% as the ISI increased from 1 ms to 6 ms. Finally, the interaction of group with ISI was also significant (F10,180 = 8.97, p < 0.001), showing that the condition of PTSD secondary to head trauma was able to affect the MEP amplitude at different ISIs. CONCLUSIONS: Our results demonstrate that PTSD can give rise to abnormalities in intracortical inhibition. Our results provide further evidence that alterations in cortical inhibitory circuits may underlie specific forms of neuroticism in humans. PMID- 15798791 TI - Selective serotonin reuptake inhibitors (SSRIs) and depression after myocardial infarction (MI). PMID- 15798792 TI - A biomaterial based approach for arsenic removal from water. AB - We demonstrate that the non-living, dried roots of the water hyacinth plant [Eichhornia crassipes(Mart.) Solms] can rapidly remove arsenic from water. Atomic absorption spectrometry was used to demonstrate that more than 93% of arsenite (As(iii)) and 95% of arsenate (As(v)) were removed from a solution containing 200 microg As l(-1) within 60 minutes of exposure to a powder produced from dried roots. No difference in removal efficiency was observed between the two oxidation states of As studied. The amount of arsenic remaining in solution was found to be less than 10 microg l(-1) which is the WHO guideline limit value for As in drinking water. The presence of arsenic in drinking water in a number of countries in the developing world has been found to be much higher than the WHO level, affecting the health of millions of people. In this paper, we show that a biomaterial produced from dried water hyacinth roots, a plant that is found in abundant supply in many parts of the world, can provide a simple, effective and yet cheap method for removing arsenic from contaminated water. PMID- 15798793 TI - Post-remediation evaluation of a LNAPL site using electrical resistivity imaging. AB - Present understanding of the earth's subsurface is most often derived from samples at discrete points (wells) and interpolations or models that interpret the space between these points. Electrical resistivity imaging techniques have produced an improved capability to map contaminants (especially NAPLs--NonAqueous Phase Liquids) away from traditional wells using actual field data. Electrical resistivity image data, confirmed by drilling, have demonstrated that LNAPL (Light NAPL--less dense than water, such as gasoline) contaminants exist outside of a delineated and remediated area in Golden, Oklahoma. The data also demonstrate that LNAPL exists between monitoring and remediation wells which indicate low contaminant levels when sampled. Additionally, the electrical images provided the drilling location with the highest concentration of hydrocarbon ever found on the site, even after two phases of remediation work had been performed, although the sampling protocols varied. The results indicate that current methods of post-remediation site characterization are inadequate for complete site characterization. PMID- 15798794 TI - Sea-urchin (Paracentrotus lividus) glutathione S-transferases and cholinesterase activities as biomarkers of environmental contamination. AB - Activities of glutathione S-transferases (GST) and cholinesterase (ChE) from Paracentrotus lividus were investigated as possible biomarkers of environmental contamination in the coastal zone. In the first phase of the study, the activity of both enzymes was determined in various tissues in order to select the most appropriate ones to be used in the following assays. In the second phase, the ChEs present in ambulacra were characterized using different substrates and selective inhibitors. In the next phase, laboratory bioassays were performed with dilutions of water-accommodated fraction of #4 fuel-oil (WAF) and benzo[a]pyrene (BaP) to determine the response of those enzymes to these pollutants and, finally, the activity of both enzymes was determined during a year in indigenous specimens from six sites on the Northwest coast of Portugal, with different pollution levels, to determine basal values and seasonal variations of ChE and GST activities. Among the several tissues tested, ambulacra and the anterior portion of the intestine were selected for ChE and GST assays, respectively. The determination of ChE in ambulacra tissue may be performed in a non-destructive way. Ambulacra ChE hydrolysed acetylthiocholine preferentially to propionylthiocholine and butyrylthiocholine and, inhibition by excess of substrate was observed. Enzymatic activity was almost fully inhibited by eserine sulfate (>98%) at concentrations equal or higher than 6.25 microM. Sensitivity to both BW284C51 (reaching 98% at 200 microM) and iso-OMPA (73% at 8 mM) was found. In laboratory bioassays, GSTs activity was inhibited by WAF and induced by BaP, whereas ChE activity was not affected by any of these environmental contaminants. Seasonal variations in enzymatic activities were found. For example, in a reference site, ChE values changed from 52.2 +/- 9.3 U mg(-1) protein in autumn to 71.8 +/- 13.3 U mg(-1) protein in spring, while GST activity changed from 129.9 +/- 29.8 U mg(-1) protein in winter to 279.0 +/- 48.0 U mg(-1) protein in autumn. Sea-urchins from reference sites presented significantly higher values of both ChE and GST than animals from contaminated sites in all seasons. In conclusion, the results of this study indicate that (i) ambulacra and the anterior portion of intestine are the most suitable tissues to measure ChE and GST activities, respectively; (ii) only one form of ChE seems to be present in ambulacra, showing properties of both typical acetylcholinesterase (AChE) and pseudocholinesterase (PChE); (iii)P. lividus GST is sensitive to both WAF and BaP even after acute exposures while ChE is not, and (iv) in spite of the significant seasonal variations observed in both enzymes in the field, P. lividus ChE and GST were capable of discriminate sites with different contamination levels and, thus, they are suitable for use as biomarkers in biomonitoring studies in the coastal zone. PMID- 15798795 TI - Solvation parameters for the 209 PCBs: calculation of physicochemical properties. AB - Descriptors in the Abraham equations for all the 209 PCBs have been obtained from literature chromatographic data. The six descriptors suffice for the prediction of water to octanol partitions, and for the prediction of various water to solvent and gas to solvent partitions, solubility in water, and water to micelle partition. For water to octanol, gas to dry octanol, gas to wet octanol, solubility of the solid PCBs in water, and gas to water processes, our predictive values agree excellently with the adjusted experimental values of Li et al. for sixteen particular PCBs, and with other experimental observations. We use our predictions to shed light on Henry's law constants of PCBs in water, which are the inverse of gas to water partitions. PMID- 15798796 TI - Relationship between different size classes of particulate matter and meteorology in three European cities. AB - Evidence on the correlation between particle mass and (ultrafine) particle number concentrations is limited. Winter- and spring-time measurements of urban background air pollution were performed in Amsterdam (The Netherlands), Erfurt (Germany) and Helsinki (Finland), within the framework of the EU funded ULTRA study. Daily average concentrations of ambient particulate matter with a 50% cut off of 2.5 microm (PM2.5), total particle number concentrations and particle number concentrations in different size classes were collected at fixed monitoring sites. The aim of this paper is to assess differences in particle concentrations in several size classes across cities, the correlation between different particle fractions and to assess the differential impact of meteorological factors on their concentrations. The medians of ultrafine particle number concentrations were similar across the three cities (range 15.1 x 10(3) 18.3 x 10(3) counts cm(-3)). Within the ultrafine particle fraction, the sub fraction (10-30 nm) made a higher contribution to particle number concentrations in Erfurt than in Helsinki and Amsterdam. Larger differences across the cities were found for PM2.5(range 11-17 microg m(-3)). PM2.5 and ultrafine particle concentrations were weakly (Amsterdam, Helsinki) to moderately (Erfurt) correlated. The inconsistent correlation for PM2.5 and ultrafine particle concentrations between the three cities was partly explained by the larger impact of more local sources from the city on ultrafine particle concentrations than on PM2.5, suggesting that the upwind or downwind location of the measuring site in regard to potential particle sources has to be considered. Also, relationship with wind direction and meteorological data differed, suggesting that particle number and particle mass are two separate indicators of airborne particulate matter. Both decreased with increasing wind speed, but ultrafine particle number counts consistently decreased with increasing relative humidity, whereas PM2.5 increased with increasing barometric pressure. Within the ultrafine particle mode, nucleation mode (10-30 nm) and Aitken mode (30-100 nm) had distinctly different relationships with accumulation mode particles and weather conditions. Since the composition of these particle fractions also differs, it is of interest to test in future epidemiological studies whether they have different health effects. PMID- 15798797 TI - Evaluation of interference to conventional and real-time PCR for detection and quantification of fungi in dust. AB - Advances in polymerase chain reaction (PCR) have permitted accurate, rapid and quantitative identification of microorganisms in pure cultures regardless of viability or culturability. In this study, a simple sample processing method was investigated for rapid identification and quantification of fungal spores from dust samples using both conventional and real-time PCR. The proposed method was evaluated for susceptibility to interference from environmental dust samples. Stachybotrys chartarum and Aspergillus fumigatus were used as test organisms. The sensitivity of detection in pure culture was 0.1 spore DNA equivalents per PCR reaction corresponding to 20 spores ml(-1) in the sample. However, 1 spore DNA equivalent per PCR reaction corresponding to 200 spores ml(-1) in the sample was the lowest amount of spores tested without interference in dust samples spiked with spores of either fungal species. The extent of inhibition was calculated using conventional and real-time PCR reactions containing fungal spores, specific primers, specific probes (for real-time PCR) and various amounts of dust. The results indicate that the extent of inhibition by dust on PCR varies with the type and amount of dust, and number of spores. No interference in the analysis of spiked samples was detected from 0.2 mg ml(-1) of four real-life dust samples at p-value >0.05 using 2 x 10(4) spores for conventional PCR and 2 x 10(5) spores for real-time PCR. However, samples containing >0.2 mg ml(-1) real-life dust compromised the PCR assay. These results suggest the potential usefulness of a simple sample processing method in conjunction with PCR for monitoring the fungal content of aerosols collected from indoor environments. PMID- 15798798 TI - Multivariate cluster analysis as a versatile tool for the quality assessment of short chain chloroparaffin quantification in environmental samples. AB - The compositions of commercially available solutions of short chain chlorinated paraffins (SCCP) and technical mixtures were determined by high resolution gas chromatography (HRGC) electron capture negative ionisation (ECNI) combined with low resolution mass spectrometry (LRMS). Differences and similarities in the relative distribution of congeners and homologues were investigated by cluster analysis. Samples could be separated into two groups/clusters with similar molecular mass, chlorination degree and congener pattern. With the exception of one sample, the use of CH4-ECNI-MS led to an overestimation of the chlorine content. Moreover, the influence of different SCCP mixtures on the quantification was studied. The results showed that small differences in the chlorine content could sometimes cause substantial systematic errors of up to 119% despite similar homologue and congener patterns. Errors of quantification corresponded to the deviations between response factors of sample and standard. As a consequence SCCPs should be quantified with a standard matching the overall response factor of the sample. PMID- 15798799 TI - Precipitation, pH and metal load in AMD river basins: an application of fuzzy clustering algorithms to the process characterization. AB - In the present work, Acid Mine Drainage (AMD) processes in the Chorrito Stream, which flows into the Cobica River (Iberian Pyrite Belt, Southwest Spain) are characterized by means of clustering techniques based on fuzzy logic. Also, pH behavior in contrast to precipitation is clearly explained, proving that the influence of rainfall inputs on the acidity and, as a result, on the metal load of a riverbed undergoing AMD processes highly depends on the moment when it occurs. In general, the riverbed dynamic behavior is the response to the sum of instant stimuli produced by isolated rainfall, the seasonal memory depending on the moment of the target hydrological year and, finally, the own inertia of the river basin, as a result of an accumulation process caused by age-long mining activity. PMID- 15798800 TI - Solvent-free sampling with di-n-butylamine for monitoring of isocyanates in air. AB - The solvent-free sampler for airborne isocyanates consisted of a polypropylene tube with an inner wall coated with a glass fibre filter, coupled in series with a 13 mm glass fibre filter. The filters were impregnated with reagent solution containing equimolar amounts of di-n-butylamine (DBA) and acetic acid. Air sampling was performed with an air flow of 0.2 l min(-1). The formed isocyanate DBA derivatives were determined using liquid chromatography and tandem mass spectrometry. The sampler was investigated in regard to collection principle and extraction of the formed derivatives with good results. The possibility to store the sampler before sampling and to perform long-term sampling was demonstrated. Field extraction of the sampler was not necessary, as there was no difference between immediately extracted samples and stored ones (2 days). In comparative studies, the sampler was evaluated against a reference method, impinger-filter sampling with DBA as reagent. The ratios between the results obtained with the sampler and the reference in a test chamber at a relative humidity (RH) of 45% was in the range of 83-109% for isocyanates formed during thermal decomposition of PUR. At RH 95%, the range was 72-101% with the exception of isocyanic acid. In two field evaluations, the ratios for fast curing 2,4'- and 4,4'-methylene bisphenyl diisocyanate (MDI) was in the range 81-113% and for the 3-ring MDI the range was 54-70%. For the slower curing 1,6-hexamethylene diisocyanate (HDI) and HDI isocyanurate, the ratios were in the range 78-145%. In conclusion, the solvent-free sampler is a convenient alternative in most applications to the more cumbersome impinger-filter sampler. PMID- 15798801 TI - Indoor air sampling of organophosphate triesters using solid phase extraction (SPE) adsorbents. AB - In this paper we demonstrate the use of SPE cartridges for sampling of organophosphate triesters in indoor air by adsorptive enrichment. The method has been optimised for the sampling and analysis of organophosphate triesters using a 25 mg aminopropyl silica SPE cartridge. The same cartridge is used for the active air sampling as well as for the subsequent extraction and clean-up of the sample. This makes the method fast and eliminates some tedious and time-consuming manual sample handling steps. Sampling and extraction efficiency was high for the investigated organophosphates, and limit of detection was in the range 0.1-0.3 ng m(-3). The method was applied to measurements of organophosphate triesters in two lecture halls and an electronics dismantling facility, and was compared with results from common filter/adsorbent sampling at each site. Analysis was made by GC with selective detection by NPD set in phosphorus mode, and by GC-MS. Thirteen organophosphate triesters (not counting isomers) were detected in the electronic dismantling facility. Chlorinated organophosphate triesters were detected in all locations with concentrations over 1 microg m(-3) in the lecture halls. This kind of adsorptive enrichment using an SPE cartridge could be adjusted to other types of analytes as well. PMID- 15798802 TI - Air quality and ventilation fan control based on aerosol measurement in the bi directional undersea Bomlafjord tunnel. AB - Aerosol, NO and CO concentration, temperature, air humidity, air flow and number of running ventilation fans were measured by continuous analysers every minute for a whole week for six different one-week periods spread over ten months in 2001 and 2002 at measuring stations in the 7860 m long tunnel. The ventilation control system was mainly based on aerosol measurements taken by optical scatter sensors. The ventilation turned out to be satisfactory according to Norwegian air quality standards for road tunnels; however, there was some uncertainty concerning the NO2 levels. The air humidity and temperature inside the tunnel were highly influenced by the outside metrological conditions. Statistical models for NO concentration were developed and tested; correlations between predicted and measured NO were 0.81 for a partial least squares regression (PLS1) model based on CO and aerosol, and 0.77 for a linear regression model based only on aerosol. Hence, the ventilation control system should not solely be based on aerosol measurements. Since NO2 is the hazardous polluter, modelling NO2 concentration rather than NO should be preferred in any further optimising of the ventilation control. PMID- 15798803 TI - Stratospheric temperature monitoring using a vibrational Raman lidar. Part 1: aerosols and ozone interferences. AB - Lidar measurements of temperature for the upper troposphere and lower stratosphere are commonly derived by the Raman technique. Lidar signals derived from vibrational Raman processes have been subjected to numerous simulation tests to examine their sensitivity to the presence of aerosols and ozone in the atmosphere. The influence of aerosols characteristics (wavelength dependence of aerosol extinction and particle phase function) and of ozone concentration on Raman temperature profiles is estimated. Simulations indicate large temperature deviations for post-volcanic conditions. For a Raman backscatter at 607 nm, bias is below 1 K for a total optical depth less than 9 x 10(-3) in the case of a stratospheric contamination and less than 6 x 10(-3) for a tropospheric contamination. The effect of aerosols depends on phase function and a few parameters such as altitude, optical depth and the shape of the high-altitude cloud. The wavelength dependence of aerosol extinction has some influence only for severe post-volcanic conditions (Scattering Ratio, SR >2). For a Raman backscatter at 387 nm, bias is larger and can be significant even in background aerosol conditions. Changes in the ozone density profile lead to significant Raman temperature deviations only for some specific conditions. Results suggest that both aerosol and ozone corrections are necessary to obtain an accuracy better than the 1 K requested for most atmospheric applications. PMID- 15798804 TI - In vivo distribution and fractionation of indium in rats after subcutaneous and oral administration of [(114m)In]InAs. AB - Two in vivo experiments were carried out in this study. In the first experiment five rats were given two subcutaneous injections of [(114m)In]InAs. Major sites of accumulation were spleen, liver and kidney. The intracellular distribution of indium was examined by differential centrifugation. The cytoplasmic fraction contained most of the indium activity followed by the mitochondrial fraction. Both outcomes are in close agreement with the results obtained in previous studies. Chromatographic separations on a preparative size exclusion column were carried out. It was shown that indium was mostly bound to high molecular mass compounds in serum and in the cytoplasmic fraction of spleen, liver and kidney. In a second experiment five rats were given four oral doses of [(114m)In]InAs over a short period. Prior to this experiment the in vitro solubility of cold InAs in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) was determined using graphite furnace atomic absorption spectroscopy. In the case of the SGF only 1.3% of an InAs suspension dissolved after 48 hours incubation at 37 degrees C. InAs was not soluble in SIF. Uptake of InAs after oral administration was minimal (<1%). Due to incomplete removal of traces of [(114m)In]InAs from the gastrointestinal tract, it was impossible to calculate accurately the in vivo distribution over the different organs. As the uptake and consequently the activity in the organs were very low, no further chromatographic separations could be carried out. Considering this very low uptake, it can be concluded that InAs will not accumulate in the body after oral exposure. PMID- 15798805 TI - Perfluorinated organic compounds in human blood serum and seminal plasma: a study of urban and rural tea worker populations in Sri Lanka. AB - Concentrations and accumulation of 13 fluorinated organic compounds (FOCs) in human sera and seminal plasma were measured in an Asian developing country, Sri Lanka. Six of the FOCs, PFOS (perfluorooctanesulfonate), PFHS (perfluorohexanesulfonate), PFUnA (perfluoroundecanoic acid), PFDA (perfluorodecanoic acid), PFNA (perfluorononanoic acid) and PFOA (perfluorooctanoic acid), were detected in all of the sera samples. Measurable quantities of two main perfluorosulfonates, PFOS and PFHS, were found in all seminal plasma samples. The detection frequency of the predominant perfluoroalkylcarboxylate, PFOA, in seminal plasma was >70%. Accumulation of PFOS in sera was significantly positively correlated with PFOA, PFHS and PFNA. Positive linear regressions were also found between PFNA and PFUnA and PFNA and PFDA suggesting that these compounds may have a similar origin of exposure and accumulation. Significantly positive associations were observed for partitioning of both PFOS and PFNA between sera and seminal plasma. The accumulation of FOCs was not significantly different in sera from Colombo (urban population) and Talawakele (rural conventional tea workers). However, the Haldummulla population (rural organic tea workers) had relatively lower exposure to FOCs compared to the other two groups, urban and rural conventional tea workers. Concentrations of FOCs in Sri Lanka were similar to those reported for industrialized countries suggesting that human exposure to such chemicals is widespread even in developing countries. The novel finding of FOCs in human seminal plasma implies that further studies are needed to determine whether long-term exposure in humans can result in reproductive impairments. PMID- 15798806 TI - Dioxins and polychlorinated biphenyls (PCDD/Fs and PCBs) in food from farms close to foot and mouth disease animal pyres. AB - To control the outbreak of foot and mouth disease, which occurred in the UK in early 2001, a large number of farm animals were slaughtered. Where it was not possible to render or landfill the carcasses, they were destroyed by burning on open pyres, with wood, coal and other materials. Uncontrolled combustion such as this is known to produce small quantities of dioxins and an investigation was made into whether, as a result of the burning, there was an elevation in the concentrations of these compounds in food produced in the areas close to the pyres. With few exceptions, concentrations of PCDD/Fs and PCBs were within the expected ranges as predicted by reference data. No accumulation over time was evident from a repeat milk sampling exercise. Where elevated concentrations of PCDD/Fs and PCBs were found in chickens and eggs, they were in samples not destined for the food chain. Elevated levels in some samples of milk from Dumfries and Galloway were not found in earlier or later samples and may have been found as a result of a temporary feeding regime. Elevated concentrations in lamb from Carmarthenshire were from very young animals which would not have entered the food chain. There was no evidence of any significant increase in dietary exposure to PCDD/Fs and PCBs as a result of the FMD pyres. PMID- 15798807 TI - Chronological trends in trace metals recorded by a tree bark pocket in Yakushima Island, Japan. AB - Bark included within the trunk of a 200-year-old Japanese cedar tree harvested in Yakushima Island, Japan, a World Natural Heritage Site located 150 km south of mainland Japan and 800 km east of Shanghai, China, was analysed for trace metals by ICP-MS providing a chronology of atmospheric pollution. The concentration of V, As and Pb in decadal sections of the bark pocket increased 30 to 50 fold from 1900-09 to 1960-69, indicating increased atmospheric deposition of these metals. The trend coincided with the establishment and expansion of heavy industries in Kyushu, Japan, resulting in locally high levels of air pollution. V, As and Pb subsequently declined, reflecting lower industrial emissions following air pollution control legislation from the late 1960's and decline in heavy industries. Ni, Cu and Zn showed a relatively small, 7 to 10 fold increase over time. Lead isotope ratios in the bark pockets shifted from about 0.84 to 0.86 for 207Pb/206Pb and from 2.04 to 2.10 for 208Pb/206Pb, showing that the origin of atmospheric lead changed over time from coal to more diverse sources. PMID- 15798808 TI - Bartonelosis (Carrion's Disease) in the pediatric population of Peru: an overview and update. AB - Bartonellosis, or Carrion's Disease, is an endemic and reemerging disease in Peru and Ecuador. Carrion's Disease constitutes a health problem in Peru because its epidemiology has been changing, and it is affecting new areas between the highland and the jungle. During the latest outbreaks, and previously in endemic areas, the pediatric population has been the most commonly affected. In the pediatric population, the acute phase symptoms are fever, anorexia, malaise, nausea and/or vomiting. The main signs are pallor, hepatomegaly, lymphadenopathies, cardiac murmur, and jaundice. Arthralgias and weight loss have also commonly been described. The morbidity and mortality of the acute phase is variable, and it is due mainly to superimposed infections or associated respiratory, cardiovascular, neurological or gastrointestinal complications. The eruptive phase, also known as Peruvian Wart, is characterized by eruptive nodes (which commonly bleed) and arthralgias. The mortality of the eruptive phase is currently extremely low. The diagnosis is still based on blood culture and direct observation of the bacilli in a blood smear. In the chronic phase, the diagnosis is based on biopsy or serologic assays. There are nationally standardized treatments for the acute phase, which consist of ciprofloxacin, and alternatively chloramphenicol plus penicillin G. However, most of the treatments are based on evidence from reported cases. During the eruptive phase the recommended treatment is rifampin, and alternatively, azithromycin or erythromycin. PMID- 15798809 TI - Medical dental prophylaxis of endocarditis. AB - Antibiotics have long been the main reason for the increase in man's longevity. Since their discovery, man has tried to reduce the level of infection by treating with antibiotics. At the same time, prophylactic use has been suggested, although this is controversial. Their routine use is not recommended, and empirical treatments at non-therapeutic doses, and indiscriminately, should be avoided, because they may become dangerous and harmful, causing among other things, the prevalence of resistant microorganisms and the eventual potentiation of an increase in morbid states. Infectious endocarditis is a systemic pathology that can start with a bacteremia, which comes either from dental procedures or/and chronic processes that already existed. Its etiopathogeny consists of a combination of bacteremia and two other factors: Cardiac injury, which can be congenital or/and acquired, and a debilitated immunological system (patients who have transplanted organs, or those who have auto-immune diseases, such as pemphigus vulgaris, systemic lupus erythematosus). The main goal is to prevent or to fight against the transient bacteremia, reducing its intensity and duration, and also to kill the bacteria in at-risk patients. In this way, infectious endocarditis can be prevented; the dental surgeon plays an important role in the prevention of this condition, which joins medical and dental aspects. This can be done by antibiotic prophylaxis. The dentist needs to be acquainted with the medical protocols of the heart health societies. PMID- 15798810 TI - Demographic and anthropometrical analysis and genotype distribution of chronic hepatitis C patients treated in public and private reference centers in Brazil. AB - Hepatitis C virus (HCV) infection is a serious public health problem, since 80% to 85% of HCV carriers develop a persistent infection that can progress into liver cirrhosis and hepatocarcinoma. Considering that the response of hepatitis C patients to combination therapy with interferon and ribavirin depends on HCV characteristics as well as on host features, we made a retrospective analysis of demographic and anthropometrical data and HCV genotype distribution of chronic hepatitis C patients treated in public and private reference centers in Brazil. The medical records of 4,996 patients were reviewed, 81% from public and 19% from private institutions. Patients' median age was 46 years, and there was a higher prevalence of male (62%) and white patients (80%). The analysis of HCV-infecting strains showed a predominance of genotype 1 (64%) over genotypes 2 and 3. The patients' mean weight was 70.6 kg, and 65% of the patients weighed less than 77 kg. Overweight and obesity were observed in 37.8% and 13.6% of the patients, respectively. Since a body weight of 75 kg or less has been considered an independent factor that significantly increases the odds of achieving a sustained virological response, the Brazilian population seems to have a more favorable body weight profile to achieve a sustained response than the American and European populations. The finding that 65% of chronic hepatitis C patients have a body weight of 77 kg or less may have a positive pharmacoeconomic impact on the treatment of genotype 1 HCV patients with weight-based doses of peginterferon. PMID- 15798811 TI - Prevalence of human Papilloma Virus in HIV-positive and HIV-negative patients in the State of Bahia: a pilot study. AB - Human Papilloma Virus (HPV) plays a central role in the development of cervical cancer. However, other coexisting factors, such as HIV infection, must be present for this to occur. We evaluated the prevalence of HPV in HIV-positive and HIV negative patients in the city of Salvador, Bahia, Brazil, and determined the most prevalent types of HPV in these patients. Fifty-five cases were selected from among patients attending three institutions providing cervical pathology services in the city of Salvador. HIV testing (Elisa/WB), HPV-DNA testing by PCR, colposcopy, cytology and biopsy were carried out in all patients. The histopathological results were classified as follows: 11 cases were normal/negative for neoplasia, 15 were diagnosed as cervical intraepithelial neoplasia grade 1 (CIN 1), 10 were CIN 2, 15 cases were CIN 3 and there were four cases of invasive squamous cell carcinoma. Among the 55 patients studied, 43 tested positive for HPV-DNA and 20 for HIV. All HIV-positive patients were positive for HPV-DNA. The most prevalent types of HPV were HPV 16, 52, 58, 53, 54, 33 and 51, and there was little difference between the groups of HIV-positive and HIV-negative patients with respect to the type of HPV encountered. The HIV positive patients were found to be infected with a greater number of types of HPV than the HIV-negative patients. This study corroborates the existence of regional variations in the distribution of certain types of HPV, which is probably due to the particular ethnic constitution found in this region of Brazil. PMID- 15798812 TI - Effect of N-acetyl-L-cysteine on lymphocyte apoptosis, lymphocyte viability, TNF alpha and IL-8 in HIV-infected patients undergoing anti-retroviral treatment. AB - N-acetyl-L-cysteine (NAC) has been proposed as an additional therapeutic agent for AIDS patients because it reduces human immunodeficiency virus type 1 (HIV-1) replication in stimulated CD4+ lymphocytes, and it ameliorates immunological reactivity. In a randomized, 180-day, double-blind, placebo-controlled trial performed with HIV-infected patients classified as A2 and A3 according to the criteria of the Center for Disease Control and Prevention, we investigated the effects of oral administration of NAC on HIV-infected patients undergoing their first anti-retroviral therapy; viral load, CD4+ lymphocyte, lymphocyte viability and apoptosis, and TNF-alpha and IL-8 levels were determined. Sixteen patients who received anti-retroviral therapy plus a placebo formed the control group and the study group consisted of 14 patients who received anti-retroviral therapy and NAC supplementation. A significant decrease was seen in viral load, TNF-alpha and IL-8 levels, and lymphocyte apoptosis, and a significant increase was found in levels of CD4+ lymphocytes and lymphocyte viability in both groups after anti retroviral treatment, but no measurable benefits of anti-retroviral therapy plus NAC oral supplementation (600 mg/day) were found in relation to anti-retroviral therapy alone, and the baseline levels of cysteine and glutathione in plasma were not recovered by this treatment. In conclusion, the daily doses of NAC necessary for the total recuperation of plasma cysteine and glutathione levels in HIV infected patients and the additional benefits following the supplementation of NAC in patients submitted to anti-retroviral therapy, need to be studied further. PMID- 15798814 TI - Serum ascorbic acid concentration in patients with acute Falciparum malaria infection: possible significance. AB - Falciparum malaria infection is associated with significant destruction of erythrocytes. This leads to the release of toxic metabolic products, including oxidant compounds. We measured the serum concentration of the antioxidant, ascorbic acid, in 129 patients presenting with acute falciparum malaria infection and in 65 healthy individuals. None of the study subjects administered any form of ascorbic acid supplementation within one week prior to participation in this study. The mean serum ascorbate concentration in infected adult males (n = 49, age range 18-50 years) was found to be 2.02 +/- 0.20 mg/dL, and it was 2.03 +/- 0.24 mg/dL in infected adult females (n = 56, age range 18-50 years). These values were significantly greater than the serum ascorbate levels (1.54 +/- 0.10 mg/dL) in healthy adult males (n = 28) and females (n = 28) (p < 0.05). In children (age range 3 to 5 years), the serum ascorbate concentration was significantly lower (1.95 +/- 0.20 mg/dL) during infection (n = 25) than in their healthy counterparts (2.9 +/- 0.24 mg/dL, n = 9) (p < 0.05). It is evident therefore that ascorbic acid plays a significant role in the pathogenesis of acute falciparum malaria in adults. Infected children also need to be given supplemental doses of ascorbate in view of the weakness of their immune system. PMID- 15798815 TI - Antibacterial effect of borage (Echium amoenum) on Staphylococcus aureus. AB - Borage (Echium amoenum) is a large annual plant of the Boraginaceae family, which grows in most of Europe and in northern Iran. The borage flower is used as a medicinal herb in France and other countries. Iranian borage is used in traditional medicine for infectious diseases, flu and as an anti-febrile. We tested the aqueous extract of borage dried flowers in vitro for its antibacterial activity. The extract showed concentration-dependent antibacterial activity against Staphylococcus aureus 8327. This activity was heat resistant, but the activity of freeze-dried extract gradually diminished during a 90-day period. The traditional use of Iranian borage flowers for infectious diseases and for controlling fever appears to be justified. PMID- 15798813 TI - Laboratory tests in the detection of extended spectrum beta-lactamase production: National Committee for Clinical Laboratory Standards (NCCLS) screening test, the E-test, the double disk confirmatory test, and cefoxitin susceptibility testing. AB - Extended spectrum beta-lactamase (ESBL) production by Klebsiella sp. and E. coli is an emerging problem. In this study, 107 clinical isolates (53 E. coli, 47 K. pneumoniae and 7 K. oxytoca) screened as ESBL producers by the NCCLS disk diffusion procedure were submitted to a double disk confirmatory test (DDT) and to the E-test double strip for confirmation of ESBL production by demonstration of clavulanic acid inhibition effect (CAIE). Only 72/107 (67%) of the isolates were confirmed as ESBL producers by DDT, with diverse results among species. By the E-test, 58/107 (54%) isolates were confirmed as ESBL producers, and 18/107 (17%) were not determinable. Susceptibility to cefoxitin was found in 57/68 (83%) of strains that did not show CAIE. ESBL detection remains a controversial issue and clinical laboratories are in need of a simple and effective way to recognize strains with this kind of resistance. PMID- 15798816 TI - Meningitis due to Enterobacter aerogenes subsequent to resection of an acoustic neuroma and abdominal fat graft to the mastoid. AB - Meningitis is an uncommon complication of neurosurgical procedures, with an incidence of 1.1% to 2.5%. Although unusual, the frequency of nosocomial Gram negative meningitis appears to be increasing. Gram-negative meningitis has been documented following disruption of the dura-arachnoid barrier secondary to trauma or surgery. The association of Gram-negative bacillary meningitis with neurosurgical procedures was first reported in the 1940's. Wolff et al. described the association between Enterobacter species and post-neurosurgical infection. More recently, risk factors for nosocomial Enterobacter meningitis have been characterized by Parodi et al. Adipose graft, as an independent risk factor has not yet been reported. A patient with acoustic neuroma resection, who developed bacterial meningitis from an abdominal fat pad graft to a mastoidectomy bed is described. A brief overview was made of post-neurosurgical Gram-negative meningitis. PMID- 15798817 TI - The importance of fluoride dentifrices to the current dental caries prevalence in Brazil. AB - Similar to that which occurred in most developed countries, dental caries have shown a significant decline in Brazil over the last two decades. Water fluoridation, expansion of preventive programs at schools, and especially, the widespread use of fluoride dentifrice are discussed as factors related to this reduction in caries. Data from epidemiological surveys and historical facts are presented to support the importance of fluoride dentifrices to the current caries prevalence in Brazil. PMID- 15798818 TI - Lack of repair of rat skull critical size defect treated with bovine morphometric protein bound to microgranular bioabsorbable hydroxyapatite. AB - The ability of a pool of bovine bone morphogenetic proteins bound to synthetic microgranular hydroxyapatite (BMPb-HA) to stimulate bone repair was determined in rat critical size defects. An 8-mm diameter defect was created in the calvaria of 25 rats. In 15 rats, the defects were filled with BMPb-HA homogenized with blood (experimental group), and in 10 rats the defects were filled only with blood clots (control). The calvariae of experimental rats were collected 1, 3 and 6 months after surgery and of the control rats at the end of surgery and 6 months thereafter. The morphometric results obtained in the radiographs showed an absence of new bone formation at 1 and 3 months post-surgery and, histologically, the defects were filled with fibrous connective tissue and numerous foci of a foreign body-type granulomatous reaction around hydroxyapatite agglomerates. At the end of 6 months, the number and size of the granulomatous foci decreased and the area of the defects was reduced by 22% compared to the 0-hour control due to the formation of new bone at their borders, although the mean area was similar to the 6-month control. We conclude that the use of BMPb-HA in the treatment of critical size bone defects of the rat skull leads to the formation of a foreign body-type granulomatous reaction that markedly inhibits new bone formation, suggesting that synthetic microgranular hydroxyapatite does not represent a good carrier for BMP-induced bone formation. PMID- 15798819 TI - Antimicrobial potential of medicaments used in healing process in dogs' teeth with apical periodontitis. AB - The objective of this study was to evaluate the influence of irrigants on the antimicrobial potential of calcium hydroxide paste in dogs' teeth with apical periodontitis. Forty-eight premolar teeth of adult mongrel dogs had their root canals opened to the oral environment for 6 months. The root canals were prepared and treated with different irrigating solutions and intracanal medicaments, according to the following groups: 1) 2.5% sodium hypochlorite (NaOCl) + calcium hydroxide paste (CHP); 2) 2% chlorhexidine (CHX) + CHP; 3) vinegar + CHP; 4) vinegar + vinegar. In group 4, both the irrigating solution and intracanal medicament were vinegar, which was renewed every 7 days. Each sample was collected by using two paper points maintained in position for 1 min, and individually transported and immersed in 7 mL of Letheen broth (LB), followed by incubation at 37 degrees C for 48 h. Microbial growth was analyzed by two methods: turbidity of the culture medium and subculture on a specific nutrient broth (brain heart infusion). After 21 days, all experimental groups had microbial growth, however, in different percentages: group 1 - 30%; group 2 - 30%; group 3 - 40%; group 4 - 60%. All materials tested had antimicrobial potential; however, the influence of calcium hydroxide paste on the control of microorganisms must be remembered. PMID- 15798820 TI - Remineralization of carious dentin. II: In vivo microradiographic and chemical studies in human permanent teeth capped with calcium hydroxide. AB - The main aim of this in vivo study was to evaluate a possible remineralization of human carious dentin by means of chemical and microradiographic studies. Eighty six samples of carious dentin were removed from 36 permanent teeth of 24 patients. These were divided into untreated (control) and chemically pure calcium hydroxide-capped (experimental) samples and analyzed at intervals varying from 10 to 120 days. They were classified according to depth of caries and degree of dentin softening and evaluated in relation to weight, phosphorus concentration, qualitative and quantitative microradiography and absolute values of total mineral content. One of two halves of each sample was selected for chemical studies and the other for total content of mineral salts. Experimental samples were examined with a light microscope and the results obtained showed a qualitative increase in radiopacity. Quantitatively, it was observed that, in the case of samples analyzed for phosphorus concentration, the average mean of differences in percentage increase after treatment was 9.6%, while for the samples evaluated microradiographically for total mineral content, it was 22.29%. In both cases, the differences were statistically significant. PMID- 15798821 TI - Influence of Er:YAG laser irradiation on apical sealing of four different sealers. AB - The sealing of the root canal system is of fundamental importance for successful endodontic treatment. To obtain an adequate apical seal, many factors must be considered such as the presence of smear layer and the sealer applied. After canal preparation, this layer must be removed because it prevents close contact between the dentinal walls and the sealing material. The goal of this study was to evaluate the sealing ability of four different sealers after smear layer removal with either 17% EDTA-T irrigation or Er:YAG laser irradiation of 46 teeth. The canals were sealed with four different sealers: Sealapex, Ketac Endo, AH Plus and N-Rickert. The method for smear layer removal did not influence apical sealing. AH Plus and N-Rickert allowed less dye leakage when compared to Sealapex and Ketac Endo. PMID- 15798822 TI - Microleakage study of three adhesive systems. AB - The purpose of this in vitro study was to evaluate the efficacy of three hydrophilic dentin adhesive systems to reduce class II restoration microleakage. A total of 60 human molar teeth were used in which two box cavities were made on the distal and mesial surfaces, with a cervical margin in dentin. These cavities were randomly divided into 3 groups (n = 40 each), according to adhesive system tested: G1: OptiBond SOLO; G2: Amalgambond Plus; G3: Etch & Prime 3.0. The cavities were restored with the composite resin Z-100. The groups were thermocycled 2000 times (5 +/- 1 degrees C and 55 +/- 1 degrees C) with a dwell time of 1 min. The teeth were then immersed in 2% methylene blue, pH 7.0, for 4 h, sectioned and observed with a stereomicroscope MEIJI 2000 (35X). The evaluation was made using scores (0-4) and the results were expressed through the sum of the ranks. G1 = 1994.00; G2 = 2294.00; G3 = 2972.00. The three groups were significantly different. The self-etching adhesive system Etch & Prime 3.0 was less effective in preventing microleakage. The OptiBond SOLO adhesive was the most effective in reducing microleakage in dentin margins when compared with Amalgambond Plus and Etch & Prime 3.0. PMID- 15798823 TI - Evaluation of depth of cure and Knoop hardness in a dental composite photo activated using different methods. AB - The aim of this study was to evaluate the depth of cure and Knoop hardness in the P60 composite resin photo-activated using different methods. A bipartite brass matrix (3 mm in diameter X 11 mm in height) was filled with the composite and photo-activation was performed using continuous light, exponential light, intermittent light, plasma arc curing (PAC) or light-emitting diodes (LED). After opening the matrix, the uncured material was removed with a steel spatula and the polymerized composite was measured using a pachymeter. The specimens were then included in self-curing acrylic resin and worn longitudinally and the hardness was measured on the surface and at depths of 1, 2, 3, 4 and 5 mm. The data were analyzed by ANOVA and Tukey's test (5%). The results showed that the depth of cure was higher with the intermittent light, followed by continuous light, exponential light, PAC and LED methods. Up to a depth of 2 mm, all methods revealed similar hardness values, but there were differences between them at other depths, at which LED demonstrated the lowest values followed by PAC. PMID- 15798824 TI - Average arch widths and associated changes between initial, post-treatment and post-retention measurements. AB - Computer image monitoring was used for evaluation of dental arch changes. A new special device captured geometrically calibrated images permitting comparison of several different dental casts. In the first part of this study 792 sets of study casts were screened. Measurements of dental arch width between reference points of canines, first premolars and first molars were made: upper jaw: men: canines: 35.1 +/- 0.13 mm; first premolars: 37.5 +/- 0.13 mm; first molars: 48.1 +/- 0.19 mm; women: canines: 33.4 +/- 0.13 mm; first premolars: 35.6 +/- 0.15 mm; first molars: 46.7 +/- 0.19 mm. In the second part of the study, changes between initial, post-treatment and post-retention alignment (5-years after orthodontic therapy) of upper and lower dental arch of 36 subjects were analyzed. Upper and lower arch compression in first premolars and molars area was visible before treatment. We conclude that computer image monitoring can be used for evaluation of dental arch changes during the different steps of treatment. PMID- 15798826 TI - Effects of aluminum-copper alloy filtration on photon spectra, air kerma rate and image contrast. AB - This study evaluated the performance of aluminum-copper alloy filtration, without the original aluminum filter, for dental radiography in terms of x-ray energy spectrum, air kerma rate and image quality. Comparisons of various thicknesses of aluminum-copper alloy in three different percentages were made with aluminum filtration. Tests were conducted on an intra-oral dental x-ray machine and were made on mandible phantom and on step-wedge. Depending on the thickness of aluminum-copper alloy filtration, the beam could be hardened and filtrated. The use of the aluminum-copper alloy filter resulted in reductions in air kerma rate from 8.40% to 47.33%, and indicated the same image contrast when compared to aluminum filtration. Aluminum-copper alloy filtration may be considered a good alternative to aluminum filtration. PMID- 15798825 TI - Reproducibility of sterilized rubber impressions. AB - Impressions, dentures and other dental appliances may be contaminated with oral micro-flora or other organisms of varying pathogenicity from patient's saliva and blood. Several approaches have been tried to control the transmission of infectious organisms via dental impressions and because disinfection is less effective and has several drawbacks for impression characterization, several sterilization methods have been suggested. This study evaluated the reproducibility of rubber impressions after sterilization by different methods. Dimensional accuracy and wettability of two rubber impression materials (vinyl polysiloxane and polyether) were evaluated after sterilization by each of three well-known methods (immersion in 2% glutaraldehyde for 10 h, autoclaving and microwave radiation). Non-sterilized impressions served as control. The effect of the tray material on impression accuracy and the effect of topical surfactant on the wettability were also evaluated. One-way ANOVA with Dunnett's method was used for statistical analysis. All sterilizing methods reduced the reproducibility of rubber impressions, although not always significantly. Microwave sterilization had a small effect on both accuracy and wettability. The greater effects of the other methods could usually be overcome by using ceramic trays and by spraying impression surfaces with surfactant before pouring the gypsum mix. There was one exception: glutaraldehyde still degraded dimensional accuracy even with ceramic trays and surfactant. We conclude that a) sterilization of rubber impressions made on acrylic trays was usually associated with a degree of dimensional change; b) microwave energy seems to be a suitable technique for sterilizing rubber impressions; c) topical surfactant application helped restore wettability of sterilized impressions. PMID- 15798827 TI - Comparison of two cleansing pastes for the removal of biofilm from dentures and palatal lesions in patients with atrophic chronic candidiasis. AB - The efficacy of 2 oral hygiene products, an experimental toothpaste specific for complete denture cleansing and a regular standard toothpaste, was compared in terms of denture biofilm removal and cure of palatal lesions in patients with atrophic chronic candidiasis. The degree of correlation between presence of biofilm and mucosa erythema was also evaluated. Twenty-four complete denture wearers (45-80 years old) were divided into 2 groups: experimental paste and standard toothpaste (Sorriso-Kolynos, Brazil). Both groups received soft toothbrushes. The internal surfaces of upper dentures were stained using 1% sodium fluorescein and photographed at a 45 masculine angle at 0, 15, 30 and 60 days. The slides were scanned and the areas of interest (denture total area and biofilm area) were measured (Image Tool software). The degree of erythema was evaluated on slides according to the Prosthesis Tissue Index. There was a significant reduction (1%) in the degree of biofilm (ANOVA/Tukey) between the two initial visits (0 and 15 days) and the two final visits (30 and 60 days), and in the average erythema scores (Kruskal-Wallis) between 0 and 60 days, in both groups. The Mann-Whitney test showed a significant difference (1%) between pastes in terms of biofilm degree, but no difference was found for the erythema score. Correlation values between biofilm and erythema degree were 0.3801 (experimental paste) and (0.3678 (standard toothpaste). We may therefore conclude that the experimental product was efficient for the removal of denture plaque biofilm. PMID- 15798828 TI - Influence of chemical and mechanical polishing on water sorption and solubility of denture base acrylic resins. AB - Influence of polishing methods on water sorption and solubility of denture base acrylic resins was studied. Eighty samples were divided into groups: Classico (CL), and QC 20 (QC) - hot water bath cured; Acron MC (AC), and Onda Cryl (ON) - microwave cured; and submitted to mechanical polishing (MP) - pumice slurry, chalk powder, soft brush and felt cone in a bench vise; or chemical polishing (CP) - heated monomer fluid in a chemical polisher. The first desiccation process was followed by storage in distilled water at 37 +/- 1 degrees C for 1 h, 1 day, 1, 2, 3 and 4 weeks. Concluding each period, water sorption was measured. After the fourth week, a second desiccation process was done to calculate solubility. Data were submitted to analysis of variance, followed by Tukey test (p 3 days before acyclovir therapy. (p = 0.01, two-tailed Fisher test). PMID- 15798869 TI - [Equine influenza in Chile (1963-1992): a possible human case]. AB - In Chile equine influenza has been strongly related to continental epizootics. First outbreak was described in 1963; main outbreaks were reported in 1977, 1985 and 1992. Equine influenza virus was isolated in three opportunities: H7N7 (A/equi/1/Santiago, Chile/1977); H3N8 (A/equi/2/Santiago, Chile/1985) and H3N8 (A/equi/2/Quillota, Chile/1992). The most serious outbreak was caused by strain H7N7 in 1977. Since 1992 equine influenza has not been reported in our country. In 1973 was described a case of influenza and seroconversion in a human being related with horses suffering a respiratory distress diagnosed as equine influenza; unfortunately the isolated virus was not typed. PMID- 15798870 TI - [Health decision making in AIDS family caregiver]. AB - AIDS infected patients receive care at home; a family member assumes the caregiver role and takes health decisions that affect the person's life. It is important to know the conflicts that the families are confronting in relation to health decision making, in order to plan intervention strategies according with their real needs. This is a descriptive study, in which a sample of 38 family caregivers of AIDS persons receiving ambulatory care at the Clinica Familia, were used for this research. An O'Connor and Jacobsen instrument was used for this study. CONCLUSIONS: the family caregivers are women, the majority of them are mothers that live with their sick son or daughter, and who had assumed the role at least for 2 years. The family caregivers have conflicts about to continuing or not caring for the sick person, and of telling others about the disease. The family caregivers are in one of the phases of the changing process. Consequently, they are receptive to helping strategies that must be based in interventions oriented to listening and supporting, more than just providing information. PMID- 15798871 TI - [Utility of stool culture in inpatient]. AB - Several studies have concluded that it is inappropriate to perform stool cultures (SC) to inpatients that have stayed hospitalized three days or more because nosocomial diarrhea is not due to enteric pathogenic bacteria that are searched through this exam. The aim of this paper was to analyze the SC yield performed to patients hospitalized in the UC Health Net since January to December 2002 and the rate of positive results obtained depending on the length of hospitalization in order to define if international guidelines are useful to the national reality. During twelve months 3474 SC were evaluated, 458 (13.2%) belonged to inpatients. Of them 16 (3.5%) were positive, 13 were obtained on the first day, 2 at the second day and 1 at the fifth day of hospitalization. Only 1/190 (0.5) SC obtained after two days of hospitalization yielded a positive result. These results reinforce the international recommendation of not performing SC to inpatients that have stayed more than three days at the hospital; besides, considering previous figures, we suggest to make a cutoff at day two of hospitalization, after two days there is no a significant clinical impact. This strategy means to reduce 41% of these analyses and to save the corresponding money. PMID- 15798872 TI - [Diagnosis, management and prevention of infections in cirrhotic patients]. AB - Infections in patients affected with liver cirrhosis are frequent, recurrent and associated to unfavorable outcome. They are facilitated by acquired and progressive defects on the innate immune and reticuloendothelial system that are aggravated by alcohol consumption. Infections in patients with cirrhosis are typically bacterial or viral in origin and have in most cases a stereotyped clinical presentation, although diagnosis may be difficult in some cases. Pneumonia, urinary tract infection, bacteremia and spontaneous bacterial peritonitis explain more than 90% of the cases. The latter requires a high clinical suspicion and a standardized diagnostic work up. Preventive strategies are important in the management of these patients and include chemoprophylaxis against spontaneous bacterial peritonitis in selected cases, vaccines against pneumococcal and influenza infections, and hepatitis A and B vaccine in susceptible patients. Due to limited seroconversion, active immunization should be applied as earlier as possible, before clinical deterioration ensues. PMID- 15798875 TI - [Leptospirosis: report of a fulminant fatal case and review]. AB - In relation to the explosive increase of new and reemerging infectious diseases that threaten the humanity, and as a result of this clinical case, a literature review is done on Leptospirosis stressing out the necessity of having in mind the leptospirosis in the differential diagnostic for fever and jaundice syndrome. PMID- 15798874 TI - [Non-typable Haemophilus influenzae severe pneumonia in an infant: case report]. AB - Haemophilus influenzae b (Hib) disease used to be a frequent cause of pneumonia in children. After the widespread use of Hib conjugate vaccines, non-b H. influenzae have become an important cause of pneumonia. We present the case of an infant with severe pneumonia with empyema and bacteremia after he had completed the Hib vaccination schedule. Non typable H. influenzae was recovered from blood and pleural effusion cultures. We review the literature about this unusual clinical presentation in our community and alert about the possible emergence of non typable H. influenzae as a frequent cause of community-acquired pneumonia in children. PMID- 15798873 TI - [Influenza: the reemergence of an ancient disease and its risk of pandemia]. AB - Influenza is a seasonally, acute respiratory disease, highly transmissible. The diversity of the natural reservoirs of influenza A virus and its faculty of reassortment increase the risk of a new pandemia. Prevention strategies during the outbreaks include vaccination indicated to risk population as infants between 6 to 2 years old, persons above 65 years old, pregnant women and patients with underlying diseases. Antiviral prophylaxis is useful to control small outbreaks and to be used in household contacts of risk population who have not been vaccinated. Antiviral drugs as a treatment should be considered in persons with severe disease. During a pandemia these prevention measures must be reinforced and rational use of antiviral drugs and vaccine with the pandemic strain should be emphasized. PMID- 15798876 TI - [Bretonneau and Louis: differentiation and characterization of typhoid fever]. PMID- 15798878 TI - Influence of pH on soil charge characteristics and cadmium sorption in some noncontaminated soils of Indian subtropics. AB - Concentrations of total dissolved cadmium (Cd) and activity of its free ions in soil solution are suggested to be influenced by soil pH, organic matter (OM) content, cation exchange capacity (CEC), and clay mineralogy. We investigated the sorption of Cd by taking 25-, 50-, and 100-microM Cd solutions in five noncontaminated soils of West Bengal, India, having differing chemical properties with batch sorption experiments. The charge characteristics and point of zero salt effect (PZSE) of all experimental soils were calculated by the potentiometric titration method measuring the adsorption of H+ and OH- on amphoteric surfaces in solutions of varying ionic strength (I). Sorption of Cd was more pronounced at pH levels greater than PZSE for all experimental soils. The CEC, OM content, clay mineralogy, and specific surface area (SSA) also had a great influence on the sorption of Cd from soil solution to soils. The relationships of Cd with those parameters were found to be consistent and the results concluded that Cd sorption in soils is strongly affected by the soil characteristics. PMID- 15798877 TI - Characterization of malignant melanoma using vibrational spectroscopy. AB - Malignant melanoma, a malignant neoplasm of epidermal melanocytes is the third most common skin cancer. In many cases, melanoma develops from nevus, which is considered as the nonmalignant stage. Fourier transform infrared microspectroscopy (FTIR-MSP), which is based on characteristic molecular vibrational spectra of cells, was used to investigate spectral differences between melanoma, nevus, and the corresponding normal epidermis. In the present work, FTIR-MSP was performed on formalin-fixed biopsies of melanoma and nevi along with the adjoining histologically normal epidermis to understand the biochemical variations from the epidermis and identify suitable parameters for differentiation of nevi from melanoma. The comparative analysis of various parameters calculated from the spectral data of the normal epidermis and the abnormal regions showed that the changes in the nucleic acids was a significant indicator of the abnormal nature of the tissues. The RNA/DNA ratio was decreased in case of both melanoma and nevus compared to the epidermis. The amide II/amide I ratio was greater for nevus and melanoma compared to the epidermis. In contrast to other organs, the analysis of carbohydrates was not found as a suitable indicator in case of malignant melanoma. Shifts in band wave number were found to be a major distinguishing feature between the melanoma and compound nevi. The present study helps in the identification of spectral features suitable for distinction of melanoma from nevus that appear similar even in FTIR spectral features and thus can pave the way for development of in vivo screening systems based on these diagnostic markers. PMID- 15798879 TI - Acute effect of gamma irradiation on gastric acid secretion and gastric mucosal integrity in the rat. AB - The effect of 3- or 6-Gray (Gy) whole-body gamma irradiation on basal and stimulated gastric acid secretion was studied in pylorus-ligated rats. Different groups of rats were irradiated with a single 3- or 6-Gy fraction and examined 7 days after irradiation. Exposure to 3-Gy fraction led to marked increase in basal (nonstimulated) gastric acid output in the 4-h pylorus-ligated rat (47.5% compared with unirradiated controls). After exposure to 6 Gy, only 18.2% increase in gastric acid output was noted compared with unirradiated controls. Under pentagastrin or histamine stimulation, gastric acid secretion in those irradiated with 3- or 6-Gy fraction was markedly reduced compared to that of unirradiated controls. Exposure to 3- or 6-Gy gamma irradiation intensified the degree of gastric mucosal injury evoked by indomethacin or 50% ethanol in a dose-dependent manner. It is concluded that in the pylorus-ligated rat model, lower doses of gamma irradiation increase basal gastric acid secretion and impair the gastric mucosal barrier with marked increase in its permeability to H+ following stimulation of acid secretion or exposure to barrier breakers. Exposure to irradiation is likely to result in failure of the parietal cell to respond to direct stimulation with histamine or pentagastrin. PMID- 15798880 TI - Clinical holistic medicine: chronic pain in internal organs. AB - Holistic medicine seems to be efficient in the treatment of chronic pain in internal organs, especially when the pain has no known cause. It is quite surprising that while chronic pain can be one of the toughest challenges in the biomedical clinic, it is often one of the simplest things to alleviate in the holistic clinic. These pains are regarded as being caused by repressed emotions and are explained as psychosomatic reactions. Using holistic medicine, the patients can often be cured of their suffering when they assume responsibility for the repressed feelings. The holistic process theory of healing states that the return to the natural (pain free) state of being is possible whenever the person obtains the resources needed for existential healing. This shift is explained by the related quality of life and life mission theories. The resources needed are "holding" or genuine care in the dimensions of awareness, respect, care, acknowledgment, and acceptance with support and processing in the dimensions of feeling, understanding, and letting go of negative attitudes and beliefs. The preconditions for the holistic healing to take place are "love" and trust. Obtaining the full trust of the patient, therefore, seems to be the biggest challenge of holistic medicine, especially when dealing with a patient in pain. PMID- 15798882 TI - Functional recovery following pertrochanteric hip fractures fixated with the Dynamic Hip Screw vs. the percutaneous compression plate. AB - The Dynamic Hip Screw (DHS) is currently the most frequently used implant for the treatment of pertrochanteric hip fractures. The Percutaneous Compression Plate (PCCP) is a recently developed, alternative device that involves minimal invasive surgery. The objective of the present study was to compare functional recovery following these two surgical procedures. A total of 76 consecutive elderly subjects (mean age and standard deviation, 80.6 +/- 5.5) following pertrochanteric hip fracture fixation were evaluated prospectively. Functional recovery was assessed 3 and 12 weeks and 2 years following surgery. Differences between groups 3 weeks postsurgery were found only in pain level during ambulation and in the weight-bearing capability of the operated extremity, which were both in favor of the PCCP. By 3 months, both groups had improved in all measures, but did not reach their preinjury level of independence. However, the PCCP group ambulated with fewer assistive devices and demonstrated better recovery of basic activities of daily living (BADL). While the majority of the subjects from both groups ambulated independently 2 years postsurgery, the PCCP group exhibited less pain during ambulation, was more independent in ADL, and required fewer assistive devices for ambulation. To summarize, the PCCP presents enhanced short- and long-term recovery of functional abilities in comparison to DHS. However, given the limited number of patients, further studies are necessary to substantiate these results. PMID- 15798881 TI - Impact of hydrophobic pollutants' behavior on occupational and environmental health. AB - This paper reviews the influence of hydrophobic pollutant behavior on environmental hazards and risks. The definition and examples of hydrophobic pollutants are given as a guide to better understand the sources of release and the media of dispersion in the environment. The properties and behavior of hydrophobic pollutants are described and their influence on environmental hazard and risk is reviewed and evaluated. The overall outcome of the assessment and evaluation showed that all hydrophobic pollutants are hazardous and risky to all organisms, including man. Their risk effects are due to their inherent persistence, bioaccumulation potential, environmental mobility, and reactivity. Their hazardous effects on organisms occur at varying spatial and temporal degrees of emissions, toxicities, exposures, and concentrations. PMID- 15798883 TI - Profiling heparan sulfate proteoglycans in ovarian carcinoma. PMID- 15798884 TI - On the possible leakage of ET-RR1 liquid waste tank: hydrological and migration modes studies. AB - The first Egyptian (ET-RR1) research reactor has been in operation since 1961 at the Egyptian Atomic Energy Authority (EAEA) Inshas site. Therefore, at present, it faces a serious problem due to aging equipment, especially those directly in contact with the environment such as the underground settling tanks of nuclear and radioactive waste. The possible leakage of radionuclides from these aging tanks and their migration to the aquifer was studied using instantaneous release. This study was done based on the geological and hydrological characteristics of the site, which were obtained from the hydrogeological data of 25 wells previously drilled at the site of the reactor[1]. These data were used to calculate the trend of water levels, hydraulic gradient, and formulation of water table maps from 1993-2002. This information was utilized to determine water velocity in the unsaturated zone. Radionuclides released from the settling tank to the aquifer were screened according to the radionuclides that have high migration ability and high activity. The amount of fission and activation products of the burned fuels that contaminated the water content of the reactor pool were considered as 10% of the original spent fuel. The radionuclides considered in this case were H-3, Sr-90, Zr-93, Tc-99, Cd-113, Cs-135, Cs-137, Sm 151, Pu-238, Pu-240, Pu-241, and Am-241. The instantaneous release was analyzed by theoretical calculations, taking into consideration the migration mechanism of the various radionuclides through the soil space between the tank bottom and the aquifer. The migration mechanism through the unsaturated zone was considered depending on soil type, thickness of the unsaturated zone, water velocity, and other factors that are specific for each radionuclide, namely retardation factor, which is the function of the specific distribution coefficient of each radionuclide. This was considered collectively as delay time. Meanwhile, the mechanism of radionuclide migration during their passage in the water body of the aquifer was the main focus of this study. The degree of water pollution in the aquifer at a point of contact with the main water body of Ismailia Canal 1000 m from the reactor site was assessed for the instantaneous release by comparing the results obtained with the regulations of the standard limit of radionuclides in drinking water. PMID- 15798885 TI - Child injury in Israel: emergency room visits to a children's medical center. AB - The object of this study was to provide data for policy making and prevention program planning in Israel. The study examined all visits to the Department of Emergency Medicine at the Schneider Children's Medical Center in 1996 (41,279 visits in total). Approximately 22.6% of the emergency room patients were admitted following injury. Most (97%) were unintentional injury. Approximately 42% of the patients were less than 4 years old and about 20% were 2 years old. In all age groups, the rate of boys was double. Approximately 92% were Jews. Despite this low rate of non-Jewish patients, however, they constituted 20% of later hospitalizations. The main injuries recorded were bruises and wounds from blunt objects, falls, motor vehicle-related accidents, and sport injuries. The most commonly injured body parts were the head and upper and lower limbs. In 82%, medical treatment was reported and 7% were hospitalized. In examining injuries over the year, there were no significant differences between the different months, but there were clusters of injuries around various holidays--bicycle and skateboard accidents at Rosh Hashanah, Yom Kippur, and Succoth; pedestrian accidents around Lag BaOmer; burns on Purim, Hannukkah, and Passover; and accidental poisoning around Passover. The findings gave an indication of the nature of the injured population groups. These data could be useful for prevention strategy, both on the level of physical injury as well as on the level of the times of the year, when the risk was higher. The data collected very strongly raise the urgent need for establishing a national surveillance system, which would allow tracking injury-related data with respect to young people throughout the country. PMID- 15798886 TI - Managing scoliosis in a young child with Rett syndrome: a case study. AB - Rett syndrome is a genetic disorder primarily affecting females. One of its most disabling features is the severe and rapid progression of scoliosis. So far, only surgical intervention has succeeded in reversing the development of scoliosis in Rett syndrome. The present study describes a new management approach implemented with a girl with Rett syndrome. The core of the management regime was intensive: asymmetrical activation of trunk muscles through equilibrium reactions. The X rays accompanying the article (evaluated by four experienced orthopedic surgeons blinded to the intervention process) suggested that the intervention was successful in reversing the progress of the scoliosis for the above-mentioned child. Discontinuation of treatment led to severe and rapid deterioration of the spinal curve. Due to the fact that this was a case study, generalization is limited, but we suggest further investigation and studies with this method. PMID- 15798887 TI - Arsenic intoxication-induced reduction of glutathione level and of the activity of related enzymes in rat brain regions: reversal by DL-alpha-lipoic acid. AB - The purpose of this study was to examine the effects of DL: -alpha-lipoic acid (LA) on arsenic (As) induced alteration of glutathione (GSH) level and of the activity of glutathione-related enzymes-glutathione peroxidase (GSH-Px), glutathione reductase (GR), and glucose-6-phosphate dehydrogenase (G6PDH)-in rat brain regions (cortex, hypothalamus, striatum, cerebellum and hippocampus). Male Wistar rats of 150+/-10 g weight were divided into four groups: control and three experimental groups supplemented with arsenic (sodium arsenite) alone (100 ppm mixed in drinking water), lipoic acid alone (70 mg kg(-1) body weight), arsenic plus lipoic acid (100 ppm arsenic in drinking water plus 70 mg lipoic acid kg(-1) body weight). The arsenic content of brain regions was found to increase with the administration of sodium arsenite. Arsenic exposure elicited a significant decline in glutathione content and in the activity of related enzymes, with the greatest decreases seen in the cortex, striatum, and hippocampus, whereas there were no significant differences between control rats and the group treated with lipoic acid alone. Highly elevated content of the thiobarbituric acid-reactive substance malondialdehyde (MDA) in the brain regions of arsenic-exposed rats reflected extensive lipid peroxidation (LPO) processes. Simultaneous lipoic acid treatment was effective in reducing brain regional arsenic levels and lipid peroxidation and in increasing the glutathione content and the activity of its related enzymes. Lipoic acid, by acting as an alternative sulfhydryl nucleophile to glutathione, prevents its oxidation to glutathione disulfide in detoxifying reactions against reactive oxygen species and consequently increases the activity of glutathione-related enzymes. PMID- 15798888 TI - Species differences in the metabolism of di(2-ethylhexyl) phthalate (DEHP) in several organs of mice, rats, and marmosets. AB - To clarify species differences in the metabolism of di(2-ethylhexyl) phthalate (DEHP) we measured the activity of four DEHP-metabolizing enzymes (lipase, UDP glucuronyltransferase (UGT), alcohol dehydrogenase (ADH), and aldehyde dehydrogenase (ALDH)) in several organs (the liver, lungs, kidneys, and small intestine) of mice (CD-1), rats (Sprague-Dawley), and marmosets (Callithrix jacchus). Lipase activity, measured by the rate of formation of mono(2 ethylhexyl) phthalate (MEHP) from DEHP, differed by 27- to 357-fold among species; the activity was highest in the small intestines of mice and lowest in the lungs of marmosets. This might be because of the significant differences between Vmax/Km values of lipase for DEHP among the species. UGT activity for MEHP in the liver microsomes was highest in mice, followed by rats and marmosets. These differences, however, were only marginal compared with those for lipase activity. ADH and ALDH activity also differed among species; the activity of the former in the livers of marmosets was 1.6-3.9 times greater than in those of rats or mice; the activity of the latter was higher in rats and marmosets (2-14 times) than in mice. These results were quite different from those for lipase or UGT activity. Because MEHP is considered to be the more potent ligand to peroxisome proliferator-activated receptor alpha involved in different toxic processes, a possibly major difference in MEHP-formation capacity could be also considered on extrapolation from rodents to humans. PMID- 15798889 TI - Effects of pesticides on human peripheral lymphocytes in vitro: induction of DNA damage. AB - Because of the widespread use of pesticides for domestic and industrial applications the evaluation of their genotoxic effects is of major concern to public health. Although various experimental data have provided evidence that pesticides can possess genotoxic properties in animals and in in vitro test systems after acute and chronic exposure, the information on the genotoxic effects of some of pesticides is limited and inconsistent. In the present study, the genotoxic potential of commonly used pesticides (i.e., dimethoate and methyl parathion from the organophosphate class, propoxur and pirimicarb from carbamates, and cypermethrin and permethrin from pyrethroids) have been evaluated. The genotoxic effects of these substances were examined using the single cell gel electrophoresis (comet) assay in freshly isolated human peripheral lymphocytes. The cells were incubated with 10, 50, 100 and 200 microg/ml concentrations of the test substances for 0.5 h at 37 degrees C and DNA damage was compared with that obtained in lymphocytes from the same donor not treated with substances. Hydrogen peroxide, 100 microM, was used as a positive control. Within the concentration ranges studied, no significant cytotoxic effects were observed. Dimethoate and methyl parathion at 100 and 200 microg/ml; propoxur at 50, 100 and 200 microg/ml, and pirimicarb, cypermethrin and permethrin at 200 microg/ml significantly increased DNA damage in human lymphocytes. PMID- 15798890 TI - Tumor necrosis factor is not required for particle-induced genotoxicity and pulmonary inflammation. AB - Particle-induced carcinogenicity is not well understood, but might involve inflammation. The proinflammatory cytokine tumor necrosis factor (TNF) is considered to be an important mediator in inflammation. We investigated its role in particle-induced inflammation and DNA damage in mice with and without TNF signaling. TNF-/- mice and TNF+/+ mice were exposed by inhalation to 20 mg m(-3) carbon black (CB), 20 mg m(-3) diesel exhaust particles (DEP), or filtered air for 90 min on each of four consecutive days. DEP, but not CB particles, induced infiltration of neutrophilic granulocutes into the lung lining fluid (by the cellular fraction in the bronchoalveolar lavage fluid), and both particle types induced interleukin-6 mRNA in the lung tissue. Surprisingly, TNF-/- mice were intact in these inflammatory responses. There were more DNA strand breaks in the BAL cells of DEP-exposed TNF-/- mice and CB-exposed mice compared with the air exposed mice. Thus, the CB-induced DNA damage in BAL-cells was independent of neutrophil infiltration. The data indicate that an inflammatory response was not a prerequisite for DNA damage, and TNF was not required for the induction of inflammation by DEP and CB particles. PMID- 15798891 TI - A life with lectins. PMID- 15798893 TI - The structure and functions of the presenilins. AB - The presenilins (PSs) were new proteins discovered in 1995 to be involved, among other functions, in the molecular mechanisms leading to Alzheimer's disease. These proteins have been the subject of many investigations since then to elucidate their molecular structures and functions. Until now, the conclusions about PS structure have been discordant, but the 8-TM structure has been accepted by the Alzheimer's community, with the evidence for the 7-TM structure largely ignored. Here the evidence is reviewed for the 6-TM, 7-TM, 8-TM and other proposed models of PS topography and possibilities offered for the differences in interpretation of the various sets of data. The conclusion is that at this stage, the 7-TM model for cell surface PS is most likely the correct one. PMID- 15798892 TI - Neurosteroid enhances glutamate release in rat prelimbic cortex via activation of alpha1-adrenergic and sigma1 receptors. AB - The present paper studied the effect and mechanism of neurosteroid pregnenolone sulfate (PREGS) on spontaneous glutamate release using electrophysiological and biochemical methods combined with a pharmacological approach. The results suggested that PREGS had a selective enhancing effect on spontaneous glutamate release in the prelimbic cortex and the hippocampus but not in the striatum. The effect of PREGS in the prelimbic cortex appeared to be via modulation of alpha1 adrenergic and sigma1 receptors, but in the hippocampus it might be dependent on sigma1 receptors only. The activation of alpha1-adrenergic receptors synergized sigma1 receptor activation in the prelimbic cortex. Intracellular calcium released from the endoplasmic reticulum, protein kinase C, adenylyl cyclase and protein kinase A played a key role in the effect of PREGS. Intracellular calcium, protein kinase C and adenylyl cyclase might be upstream events in the activation of protein kinase A after PREGS. PMID- 15798894 TI - Phosphodiesterase: overview of protein structures, potential therapeutic applications and recent progress in drug development. AB - Phosphodiesterases (PDEs) are essential regulators of cyclic nucleotide signaling with diverse physiological functions. Because of their great market potential and therapeutic importance, PDE inhibitors became recognized as important therapeutic agents in the treatment of various diseases. Currently, there are seven PDE inhibitors on the market, and the pharmacological and safety evaluations of many drug candidates are in progress. Three-dimensional (3D) structures of catalytic domains of PDE 1, -3, -4, -5 and -9 in the presence of their inhibitors are now available, and can be utilized for rational drug design. Recent advances in molecular pharmacology of PDE isoenzymes resulted in identification of new potential applications of PDE inhibitors in various therapeutic areas, including dementia, depression and schizophrenia. This review will describe the latest advances in PDE research on 3D structural studies, the potential of therapeutic applications and the development of drug candidates. PMID- 15798895 TI - The chemistry and biology of inhibitors and pro-drugs targeted to glutathione S transferases. AB - The cytosolic glutathione S-transferases are a family of structurally homologous enzymes with multiple functions, including xenobiotic detoxification, clearance of oxidative stress products, and modulation of cell proliferation and apoptosis signaling pathways. This wide-ranging functional repertoire leads to several possible therapeutic uses for isoform-specific GST inhibitors. These inhibitors may be used, in principle, to modulate tumor cell drug resistance, as sensitizers to therapeutically directed oxidative stress, to enhance cell proliferation and to augment anti-malarial drugs. With increasing knowledge of GST structural and function, rational design strategies and mechanism-based inhibitors have been exploited successfully. However, design of isoform specificity remains a significant challenge in GST inhibitor development. Strategies for further inhibitor design and their possible limitations, along with potential therapeutic uses, are summarized. PMID- 15798896 TI - Oxidation and tyrosine phosphorylation: synergistic or antagonistic cues in protein tyrosine phosphatase. AB - Protein tyrosine phosphatases (PTPs) have been generally recognised as key modulators of cell proliferation, differentiation, adhesion and motility. During signalling, several PTPs undergo two posttranslational modifications that greatly affect their enzymatic activity: tyrosine phosphorylation and cysteine oxidation. Although these modifications share their reversibility depending on the intracellular environment, their effects on enzymatic activity are opposite, tyrosine phosphorylation being correlated to enzyme activation and thiol oxidation to complete inactivation. Several papers have suggested that both these modifications occur in response to the same stimuli i.e. cell proliferation induced by numerous growth factors and cytokines. Conversely, the possibility that these two regulation mechanisms act simultaneously on PTPs has not been established and very few reports investigated this dual regulation of PTPs. To underline the relevance of the question, we discuss several possibilities: (i) that tyrosine phosphorylation and cysteine oxidation of PTPs may share the same target molecules but with different kinetics; (ii) that PTP phosphorylation and oxidation may take place on different subcellular pools of the same protein and (iii) that these two modifications, although having divergent effects on enzyme activity, cooperate in the integrated and coordinated function of PTPs during receptor tyrosine kinase signalling. We believe that our perspective will open new perspectives on an ancient problem--the apparent contradiction of opposing enzymatic regulation of many PTPs--thus clarifying their role as positive or negative transducers (or both) of many extracellular stimuli. PMID- 15798897 TI - Assessment of teeth as biomarkers for skeletal fluoride exposure. AB - Skeletal fluorosis and dental fluorosis are diseases related to fluoride (F) ingestion. Bone is the largest storage site of F in our body. Therefore, bone F concentrations are considered biomarkers for total F body burden (exposure). However, difficult accessibility limits its use as a biomarker. Thus, a more accessible tissue should be considered and analyzed as a biomarker for total F body burden. The objective of this study, which was divided into two parts, was to evaluate teeth as a biomarker for skeletal F exposure. In part 1 of the study, 70 mice of three different strains (SWR/J, A/J and 129P3/J) were exposed to different levels of water fluoridation (0, 25, 50 and 100 ppm). Bone (femora and vertebrae) and teeth from these mice were then analyzed for F concentration using Instrumental Neutron Activation Analysis (INAA). In part 2 of the study, human teeth (enamel and dentin) and bone from 30 study subjects were collected and analyzed for F concentration using INAA. Study subjects lived in areas with optimum levels of water fluoridation (0.7 and 1 ppm) and underwent therapeutic extraction of their unerupted third molars. The values of bone and teeth F concentration were correlated for parts 1 and 2 of this study. The results showed that in the animal model, where animals were exposed to a wide range of F in their drinking water, tooth [F] correlated with bone [F]. However, no correlation was seen between bone and enamel F concentrations or between bone and dentin F concentrations in the human samples. Therefore, teeth are not good biomarkers for skeletal F exposure in humans when exposure is confined to optimum levels of F in the drinking water. PMID- 15798898 TI - Reduction of abdominal wall blood flow by clamping or carbon dioxide insufflation increases tumor growth in the abdominal wall: an experimental study in rats. AB - BACKGROUND: We have previously demonstrated that there is a reduction of blood flow in the abdominal wall in rats insufflated with air concomitant with an increase in tumor growth. The present study was designed to examine whether a reduction of blood flow achieved by clamping or insufflation with carbon dioxide (CO(2)) would increase tumor growth in the abdominal wall. METHODS: In the first part of the experiments, laser Doppler blood flow of both rectus muscles was measured in 16 Wistar Fu rats. The left rectus muscle was clamped to reduce blood flow, and 5 x 10(4) adenocarcinoma cells were injected into both rectus muscles. Clamping was maintained for 45 min. In the second part, 22 rats had 5 x 10(4) adenocarcinoma cells injected into the rectus muscle and blood flow was measured. The experimental group (n = 11) was insufflated with CO(2) at 10 mmHg for 45 min; the control group (n = 11) was not insufflated. After 9 days, tumor weight and volume were analyzed. RESULTS: Clamping caused a 69% reduction of blood flow (p < 0.001), whereas no reduction was registered on the nonclamped side. Tumor weight (p = 0.028) and volume (p = 0.030) were increased on the clamped side. The insufflation of CO(2) caused a 71% reduction of blood flow, whereas no reduction was registered in the control group. Tumor weight (p = 0.006) and volume (p = 0.006) were increased in the insufflated group. CONCLUSION: Clamping, as well as CO(2) insufflation, causes a significant reduction of blood flow in the abdominal wall, which seems to increase tumor growth at the same site. PMID- 15798899 TI - Impact of hospital case volume on short-term outcome after laparoscopic operation for colonic cancer. AB - BACKGROUND: High hospital case volume has been associated with improved outcome after open operation for colorectal malignancies. METHODS: To assess the impact of hospital case volume on short-term outcome after laparoscopic operation for colon cancer, we conducted an analysis of patients who underwent laparoscopic colon resection within the COlon Cancer Laparoscopic or Open Resection (COLOR) trial. RESULTS: A total of 536 patients with adenocarcinoma of the colon were included in the analysis. Median operating time was 240, 210 and 188 min in centers with low, medium, and high case volumes, respectively (p < 0.001). A significant difference in conversion rate was observed among low, medium, and high case volume hospitals (24% vs 24% vs 9%; p < 0.001). A higher number of lymph nodes were harvested at high case volume hospitals (p < 0.001). After operation, fewer complications (p = 0.006) and a shorter hospital stay (p < 0.001) were observed in patients treated at hospitals with high caseloads. CONCLUSIONS: Laparoscopic operation for colon cancer at hospitals with high caseloads appears to be associated with improved short-term results. PMID- 15798901 TI - Identification of a cytoskeleton-bound form of phospholemman with unique C terminal immunoreactivity. AB - Phospholemman (PLM) is a 72-amino acid transmembrane protein thought to function in Na,K-ATPase regulation or assembly, similar to other members of the FXYD family of proteins. Unique to PLM among these regulatory proteins are sites for C terminal phosphorylation by PKA and PKC, although a role for phosphorylation in PLM function remains unclear. To study PLM phosphorylation, we used PLM phosphopeptides to generate antibodies to specifically detect phosphorylated PLM. Peptide affinity chromatography isolated two populations of antibodies: one reacting with standard PLM, a collection of closely-spaced 15-kDa protein bands by SDS-PAGE. About 20% of PLM antibodies reacted specifically with a single distinct form of PLM. Levels of this second immunological form (PLM-b) were increased with overexpression of PLM cDNA, and also reacted with a monoclonal antibody against the PLM N-terminus. In complete contrast to standard PLM, however, PLM-b was quantitatively insoluble in nonionic detergents and was released from tight binding by colchicine. Antibodies to PLM-b were present in two different antisera raised to the phosphorylated C-terminal peptide (residues 57-70), but not in antiserum raised to the non-phosphorylated C-terminal peptide. Despite an apparent relationship between PLM-b and phosphorylated PLM, PLM-b levels were not affected by treatment of heart cells with isoproterenol. PLM-b appears to represent a cytoskeleton-attached detergent-insoluble form of PLM with distinctive C-terminal immunoreactivity that might have implications for PLM structure and function. PMID- 15798900 TI - Biogenesis of CFTR and other polytopic membrane proteins: new roles for the ribosome-translocon complex. AB - Polytopic protein biogenesis represents a critical, yet poorly understood area of modern biology with important implications for human disease. Inherited mutations in a growing array of membrane proteins frequently lead to improper folding and/or trafficking. The cystic fibrosis transmembrane conductance regulator (CFTR) is a primary example in which point mutations disrupt CFTR folding and lead to rapid degradation in the endoplasmic reticulum (ER). It has been difficult, however, to discern the mechanistic principles of such disorders, in part, because membrane protein folding takes place coincident with translation and within a highly specialized environment formed by the ribosome, Sec61 translocon, and the ER membrane. This ribosome-translocon complex (RTC) coordinates the synthesis, folding, orientation and integration of transmembrane segments across and into the ER membrane. At the same time, RTC function is controlled by specific sequence determinants within the nascent polypeptide. Recent studies of CFTR and other native membrane proteins have begun to define novel variations in translocation pathways and to elucidate the specific steps that establish complex topology. This article will attempt to reconcile advances in our understanding of protein biogenesis with emerging models of RTC function. In particular, it will emphasize how information within the nascent polypeptide is interpreted by and in turn controls RTC dynamics to generate the broad structural and functional diversity observed for naturally occurring membrane proteins. PMID- 15798902 TI - Selection of 2'-fluoro-modified RNA aptamers for alleviation of cocaine and MK 801 inhibition of the nicotinic acetylcholine receptor. AB - The nicotinic acetylcholine receptor (nAChR) belongs to a group of five structurally related proteins that regulate signal transmission between approximately 10(12) cells of the mammalian nervous system. Many therapeutic agents and abused drugs inhibit the nAChR, including the anti-convulsant MK-801 and the abused drug cocaine. Many attempts have been made to find compounds that prevent inhibition by cocaine. Use of transient kinetic techniques to investigate the inhibition of the receptor by MK-801 and cocaine led to an inhibition mechanism not previously proposed. The mechanism led to the development of combinatorially synthesized RNA ligands that alleviate inhibition of the receptor. However, these ligands are relatively unstable. Here we determined whether much more stable 2'-fluoro-modified RNA ligands can be prepared and used to study the alleviation of receptor inhibition. Two classes of 2'-fluoro modified RNA ligands were obtained: One class binds with higher affinity to the cocaine-binding site on the closed-channel form and, as predicted by the mechanism, inhibits the receptor. The second class binds with equal or higher affinity to the cocaine-binding site on the open-channel form and, as predicted by the mechanism, does not inhibit the receptor, and does alleviate cocaine and MK-801 inhibition of the nAChR. The stability of these 2'-fluoro-RNAs expands the utility of these ligands. PMID- 15798903 TI - Inward-rectifier K+ current in guinea-pig ventricular myocytes exposed to hyperosmotic solutions. AB - Superfusion of heart cells with hyperosmotic solution causes cell shrinkage and inhibition of membrane ionic currents, including delayed-rectifer K+ currents. To determine whether osmotic shrinkage also inhibits inwardly-rectifying K+ current (I(K1)), guinea-pig ventricular myocytes in the perforated-patch or ruptured patch configuration were superfused with a Tyrode's solution whose osmolarity (T) relative to isosmotic (1T) solution was increased to 1.3-2.2T by addition of sucrose. Hyperosmotic superfusate caused a rapid shrinkage that was accompanied by a negative shift in the reversal potential of Ba(2+)-sensitive I(K1), an increase in the amplitude of outward I(K1), and a steepening of the slope of the inward I(K1)-voltage (V) relation. The magnitude of these effects increased with external osmolarity. To evaluate the underlying changes in chord conductance (G(K1)) and rectification, G(K1)-V data were fitted with Boltzmann functions to determine maximal G(K1) (G(K1)max) and voltage at one-half G(K1)max (V(0.5)). Superfusion with hyperosmotic sucrose solutions led to significant increases in G(K1)max (e.g., 28 +/- 2% with 1.8T), and significant negative shifts in V(0.5) (e.g., -6.7 +/- 0.6 mV with 1.8T). Data from myocytes investigated under hyperosmotic conditions that do not induce shrinkage indicate that G(K1)max and V(0.5) were insensitive to hyperosmotic stress per se but sensitive to elevation of intracellular K+. We conclude that the effects of hyperosmotic sucrose solutions on I(K1) are related to shrinkage-induced concentrating of intracellular K+. PMID- 15798904 TI - Opposite Cx32 and Cx26 voltage-gating response to CO2 reflects opposite voltage gating polarity. AB - Previous studies have shown that the V(j)-dependent gating behavior of gap junction channels is altered by CO(2) exposure. V(j)-dependent channel closure is increased by CO(2) in some connexin channels and decreased in others. Since the former type of channels gate on the relatively negative side by V(j) (negative gaters) and the latter at the positive side (positive gaters), it has been hypothesized that gating polarity determines the way CO(2) affects V(j) closure. To test this hypothesis, we have studied the CO(2)-mediated changes in V(j) gating in channels made of Cx32, Cx26, or a Cx32 mutant (Cx32-N2D) in which asparagine (N) at position 2 was replaced with aspartate (D). With exposure to CO(2), Cx32 channels (negative gaters) show increased V(j)-dependent closure, whereas Cx26 channels (positive gaters) respond in the opposite way to V(j). Additionally, Cx32-N2D channels (positive gaters) show decreased V(j) closure with exposure to CO(2). The reciprocal Cx26 mutant, Cx26-D2N (negative gater), could not be tested because it did not express functional homotypic channels. The data support the hypothesis that polarity of fast V(j) gating determines whether CO(2) increases or decreases the V(j) dependent closure of gap junction channels. PMID- 15798905 TI - Formic acid solubilization of marine biological tissues for multi-element determination by ETAAS and ICP-AES. AB - A simple, fast method is described for the determination of Ag, As, Cd, Cu, Cr, Fe, Ni, and Se in marine biological tissues by electrothermal atomic-absorption spectrometry (ETAAS) and Na, Ca, K, Mg, Fe, Cu, and Mn by inductively coupled plasma-atomic emission spectrometry (ICP-AES). Solubilization of the biological tissue was achieved by using formic acid with vortex mixing followed by heating to 50 degrees C in an ultrasonic bath. Once solubilized, the tissues were diluted to an appropriate volume with water for analysis. Aliquots were sampled into a graphite furnace and ICP-AES using a conventional autosampler. The method was validated by use of biological certified reference materials from NRC, DORM-2, DOLT-2, DOLT-3, LUTS-1, TORT-2, and NIST SRMs 1566b and 2976. Simplicity and reduced sample-preparation time prove to be the major advantages to the technique. PMID- 15798907 TI - Underpotential deposition of tin(II) on a gold disc electrode and determination of tin in a tin plate sample. AB - This work describes a study of the underpotential deposition (UPD) of Sn2+ on a polycrystalline gold disc electrode using cyclic voltammetry (CV) and chronocoulometry (CC). Sn2+ ions showed well-defined peaks from UPD and UPD stripping (UPD-S) in 1 mol/L HCl solutions, while bulk deposition (BD) and BD stripping (BD-S) of the ions were also observed. The measured UPD shifts, DeltaE(UPD), between the UPD-S and the BD-S peaks were more than 200 mV. The UPD charge and the surface coverage of tin were measured by CC. A new method for determining Sn2+ was therefore developed, based on the excellent electrochemical properties of the Au/Sn UPD system. A plot of the UPD-DPASV (differential pulse anodic stripping voltammetry) signal versus the Sn(II) concentration was obtained for [Sn(II)] of 1.98x10(-7) to 3.64x10(-5) M. The method developed here has been applied to determine the tin in a tin plate sample. PMID- 15798906 TI - A microwave-assisted microassay for lipases. AB - A quick and efficient two-step assay for monitoring and screening lipase activity that uses a microtitre plate is described. PMID- 15798908 TI - Osteoporosis fracture liaison experience: the Belfast experience. PMID- 15798909 TI - Transplacental passage of a nonionic contrast agent. AB - We report the presence of iopromide in the bowel and urine of a preterm infant, born 10 days after intravenous administration of the nonionic monomer to his mother. Excessive urinary iodine excretion and borderline hyperthyrotropinaemia were observed in the infant. Moreover, crossing of the fetal blood-brain barrier was demonstrated by detection of the angiographic material in CSF and thus direct fetal neurotoxic effects cannot be excluded. CONCLUSION: These widely used contrast media may cross the placenta and accumulate in various fetal tissues in significant amounts causing possible neonatal toxicity. Therefore the perinatal safety of these diagnostic agents should at least be questioned. PMID- 15798910 TI - Spontaneous atlantoaxial subluxation as a presenting manifestation of juvenile ankylosing spondylitis in a female HLA-B27-negative patient. PMID- 15798911 TI - Pharmacodynamics of chloral hydrate in former preterm infants. AB - The aim of this study was to document the pharmacodynamics of chloral hydrate in former preterm infants at term post-conception age. The degree of sedation (COMFORT), feeding behaviour and cardiorespiratory events (bradycardic events, apnoeas) before and after administration of chloral hydrate (oral, 30 mg/kg) were prospectively evaluated in former preterm infants during procedural sedation. Characteristics at birth, during neonatal stay and at inclusion were collected. Paired Wilcoxon and McNemar tests were used to study the impact of chloral hydrate. Characteristics of infants who displayed severe bradycardic events were compared to infants in whom no bradycardic events were recorded (Mann Whitney U, Fischer's exact). A significant increase in sedation (decrease COMFORT scale) was observed up to 12 h after administration. There was a minor but significant decrease in oral intake (161 to 156 ml/kg/day, P < 0.01). A significant increase in the number of bradycardic events (<80/min: 38 to 82 events, of which < 70/min: 30 to 79 of which < 60/min: 15 to 45; at least P < 0.01) and in the duration of the most severe bradycardic event (8-12.5 s) was observed. Therefore, further inclusion was stopped when 26 neonates were included. Infants who displayed severe bradycardic (< 60/min) events ( n = 13) after administration of chloral hydrate had a lower gestational age at birth without difference in post conception age at inclusion. CONCLUSION: Chloral hydrate was associated with an increase in unintended side-effects in former preterm infants, likely reflecting population specific pharmacodynamics and kinetics of chloral hydrate. PMID- 15798912 TI - Effect of ethane-1-hydroxy-1,1-bisphosphonate (EHBP) on endochondral ossification lesions induced by a lethal oral dose of uranyl nitrate. AB - A 350 mg/kg body weight (b.w.) oral dose of uranyl nitrate (UN) caused 100% mortality in mice three days after administration, due to resulting kidney lesions. Mortality decreased 50% after an oral (o) or subcutaneous (sc) dose of bisodic etidronate (EHBP). Given that bone is also a target organ for uranium (U) in acute intoxication, the aim of this work was to study the effect of exposure to a lethal oral dose of UN on endochondral ossification, and the latter's response to EHBP treatment. One hundred male Balb/c mice weighing 25 g were assigned to one of ten groups. Group I served as control. Group II received a lethal 350 mg/kg b.w. oral dose of UN by gavage. Groups III, IV, VII, and VIII received an equal dose of UN immediately followed by a single 500 mg/kg oral dose of EHBP in groups III and VII and a single 50 mg/kg subcutaneous dose of EHBP in groups IV and VIII. Groups V and IX only received a single 500 mg/kg oral dose of EHBP, and groups VI and X received a single 50 mg/kg subcutaneous dose of EHBP. The animals in groups II, III, IV, V, and VI were sacrificed 48 h after the onset of the experiment, whereas those in groups VII, VIII, IX, and X were killed at 14 days. Histologic and histomorphometric studies were performed on the femurs to determine growth cartilage width, bone volume, and metaphyseal bone activity. Our results showed that all growth cartilage and metaphyseal bone histomorphometric parameters were significantly lower in animals exposed to UN at 48 h than in controls. EHPB administration was found to prevent this condition at 48 h reaching similar values to those of controls. Although histomorphometric values did not reach control values at 14 days, they were higher than those of animals exposed to UN at 48 h not treated with EHBP. It is noteworthy that these values also decreased in animals only receiving EHBP at 14 days. Our results show that EHBP effectively ameliorates the adverse effects of a lethal dose of UN on endochondral ossification. PMID- 15798913 TI - L-Carnitine halts apoptosis and myelosuppression induced by carboplatin in rat bone marrow cell cultures (BMC). AB - Carboplatin (CP), a second generation platinum compound, is effective against various types of cancers, producing less nephrotoxicity and ototoxicity but more myelotoxicity than cisplatinum. CP-myelosuppression is the rate-limiting step of its clinical use. Prevention of CP-myelosuppression is a major target in the field of chemotherapy. Therefore, the present study investigates the use of L carnitine (LCR)-an antioxidant, cardioprotective, neuroprotective, and immunostimulant nontoxic natural compound-to protect against CP-induced myelosuppression. The viability of BMC was studied using a trypan blue exclusion technique following incubation with CP and/or LCR as a function of time and concentration. Apoptosis was tested for by detecting the amount of DNA fragmentation and the visualization of DNA ladders upon gel electrophoresis. Bone marrow progenitor cell function was examined by colony forming unit assay. Cellular contents of glutathione (GSH) and malondialdehyde (MDA) were also estimated. Results revealed that LC50 of CP is 4.7 mM and the highest safe concentration of LCR is 5 mM. Co-exposure of LCR+CP rescued BMC viability by 37% compared to the CP-treated cultures. The LCR halts CP-induced apoptosis and it significantly improves the function of the bone marrow progenitors by increasing the number of colony-forming units as a response to granulocyte/macrophage colony stimulating factors. Finally, LCR restores CP-induced GSH depletion and prevents MDA elevation in BMC. In summary, the results suggest that LCR is able to protect against CP-induced myelosuppression, which suggests its use as an adjuvant therapy. This finding merits further investigation into the mechanism(s) of such protection as well as its interaction with CP antitumor activity. PMID- 15798916 TI - Practical effects of educating nurses on the Natural Death Act. AB - BACKGROUND: The aim of this study was to investigate a nursing staff's knowledge of the Natural Death Act before and after an educational intervention. PATIENTS AND METHODS: This was a quasi-experimental designed study. Questionnaires were sent to 135 nurses in a medical center who volunteered to participate in the educational program. Demographic data and knowledge of the Natural Death Act were collected. The return rate was 85.37% (n=105). The educational program lasted for 6 h and included five subjects: hospice-palliative care, Natural Death Act, how to face suffering patients, nursing for agonized patients, and communicating with families. The program was a combination of lectures, case discussions, evaluation of individuals' values, and attitudes toward patient death. MAIN RESULTS: At baseline, the average total score in the 18-item questionnaire on the Natural Death Act was 12.96, with a range of 5-18. After the educational intervention, the average total score increased from 12.9 correct answers to 17.04 (of 18). The average increase was 4.08 points, a statistically significant difference. CONCLUSIONS: A provision of appropriate training for medical professionals appears to be a useful educational strategy, and this result shows that the Natural Death Act should be included in nursing school educational program. In the future, more effort should be made to fulfill patients' expectations and to follow medical ethics guidelines. PMID- 15798915 TI - The comparative effects of povidone-iodine and normal saline mouthwashes on oral mucositis in patients after high-dose chemotherapy and APBSCT--results of a randomized multicentre study. AB - Antimicrobial solutions are widely used in the nursing care of chemotherapy induced oral mucositis (OM). There is little evidence, however, supporting their use for reducing mucosal damage. In our study, 132 patients were randomized to use normal saline (n=65) or povidone-iodine diluted 1:100 (n=67) mouthwashes for OM prophylaxis and treatment after high-dose chemotherapy comprising BEAM or HD-L PAM followed by autologous peripheral stem cell transplantation. The study groups were well balanced in respect of age, sex, chemotherapy and the number of CD34+ cells in the graft. No significant difference was found between the groups in respect of OM characteristics, fever of unknown origin (FUO) and other infections. The antimicrobial solution was less tolerable for patients. OM occurred significantly more often in females than in males (86% vs 60%, P=0.0016) and was worse and of longer duration. The mechanical effect of mouthwashes might have a certain importance in FUO prevention. When indicating oral rinses, the patient's individual preference and tolerance of solutions offered should be considered. PMID- 15798914 TI - Cardiac toxicity assessment in locally advanced breast cancer treated neoadjuvantly with doxorubicin/paclitaxel regimen. AB - BACKGROUND: The psychological difficulty of accepting a mastectomy for locally advanced breast cancer (LABC) justifies the use of chemotherapy as neoadjuvant primary treatment. The aim of this prospective study was to assess the efficacy of the doxorubicin/paclitaxel (AT) schedule neoadjuvantly administered in terms of response rates and survival in patients with LABC, with a special focus on cardiac toxicity. PATIENTS AND METHOD: All patients were treated by doxorubicin (60 mg/m2 i.v.) bolus followed by paclitaxel (200 mg/m2) as a 3-h infusion. Treatment was repeated every 3 weeks for four or six courses and followed by surgery, radiotherapy, and hormonotherapy for patients with positive hormonal receptors. Patients with significant cardiovascular history or ECG abnormalities were not eligible for the study. Measurements of left ventricular ejection fraction (LVEF) were performed at baseline and at the end of chemotherapy. RESULTS: From 1998 to 2001, 34 consecutive patients followed up in our institution were entered into this study. Median age was 49 years (range, 32-68 years). Seventeen patients had stage IIB, 5 patients stage IIIA, and 12 patients stage IIIB disease. Twenty-one patients underwent conservative surgery, 7 radical surgery, and 6 patients no surgery due to metastatic disease occurring during treatment. An objective clinical response was noted in 22 (65%) of 34 patients (6 patients with histological complete response, 10 patients with rare malignant cells, and 6 patients with a partial response), 6 patients presented a progressive disease, and 8 patients a stable disease. Twenty-four patients have kept normal cardiac function, 7 patients had a cardiac toxicity as defined by the institution [4 (24%) of 17 patients received 360 mg/m2 of doxorubicin (A), 2 of 4 presented congestive heart failure (CHF), and 3 (21%) of 14 patients received 240 mg/m(2) of A without CHF]. Three patients did not receive four or six cycles as initially planned due to the progressive disease during the chemotherapy courses. These patients were excluded from the final analysis, particularly cardiac toxicity analysis. At time of median follow-up (42 months), 28 of 34 patients were alive (one death due to CHF, five others due to progressive disease). CONCLUSION: The AT regimen in neoadjuvant treatment for LABC remains efficient, but cardiac toxicity reported in this study underlies the necessity to optimize the schedule of AT combination. PMID- 15798917 TI - Stability and compatibility of binary mixtures of morphine hydrochloride with hyoscine-n-butyl bromide. AB - The aim of this study was to determine the compatibility and stability of morphine hydrochloride and hyoscine-N-butyl bromide combined in solution at three different concentrations and stored in polypropylene syringes at 4 degrees C and 25 degrees C over a period of 15 days. The doses assayed were 20, 60 and 120 mg/day for morphine hydrochloride and 40, 60 and 80 mg/day for hyoscine-N-butyl bromide. These dose ranges were chosen according to daily practice. At both temperatures, all mixtures can be considered as physically compatible since no evidence of incompatibility-that is precipitation, turbidity, colour change or opacity and gas production-were observed. After 15 days of storage, the percentages of hyoscine-N-butyl bromide remaining in the drug mixtures tested ranged between 96.07% and 92.23%. At the end of the study, the percentages of morphine hydrochloride remaining in the drug mixtures were 100% at both temperatures. PMID- 15798918 TI - Assessing task "burden" of daily activities requiring upper body function among women following breast cancer treatment. AB - GOALS OF WORK: To determine which individual or groups of "upper-body" daily tasks are considered most burdensome to women following breast cancer treatment, and to assess whether certain patient or treatment characteristics influence task burden. PATIENTS AND METHODS: A convenience sample of breast cancer survivors (n =619) completed a self-administered questionnaire regarding 48 daily tasks requiring upper-body function. Women were asked to rate how frequent and physically demanding each task was using a five-point Likert scale, and the product of task frequency and physical demand determined overall task burden. Tasks were ranked to identify the most burdensome individual tasks, while a factor analysis was performed to define independent constructs (groupings) among the tasks. Multiple linear regression models were fitted to consider the independent influences on task groups of various participant characteristics. MAIN RESULTS: Factor analysis identified seven distinct task groups and the individual tasks considered most burdensome fell in five of these groups, specifically whole body, flexibility, carrying/upper-body strength, hand and weighted flexion tasks. Having lymphoedema or poor fitness was associated with upper-body disability involving all seven task groups, whereas other patient and treatment characteristics were related only to certain types of activities. CONCLUSIONS: Breast cancer survivors report difficulty with a range of upper-body tasks, particularly if they also have lymphoedema or poor fitness. Using all or some of the tasks within the reported constructs in a questionnaire format, or the functional requirements of the most burdensome tasks to develop more objective and quantitative measures, would provide a solid base for the measurement of upper-body function in women with breast cancer. PMID- 15798919 TI - [Femoral neck prosthesis in the case of femoral neck fracture and ipsilateral long stem-knee tumour prosthesis]. AB - Femoral neck fracture in the case of a previously implanted ipsilateral tumour knee-prosthesis represents a challenging situation because of the long proximal stem of the knee implant. Avoiding total femur replacement, we implanted a femoral neck prosthesis (Eska Cut 2000) in a 65 year old patient to preserve as much of the femur free of implant as possible, minimizing the risk of a periprosthetic or interprosthetic fracture. At 2.5 years postoperatively, there were no signs of loosening, migration or periprosthetic fracture with a good functional result. In our case, the femoral neck prosthesis avoided the implantation of a total femur replacement. Nevertheless, this prosthesis must be considered critically and cannot be recommended as the standard implant for femoral neck fracture in the case of an ipsilateral long stem-knee prosthesis. PMID- 15798920 TI - Heat shock during early somitogenesis induces caudal vertebral column defects in Atlantic salmon (Salmo salar). AB - In several terrestrial vertebrates, heat shock (HS) during somitogenesis causes vertebral deformities. To determine if vertebral deformities can occur due to sudden temperature changes during early development in fish, Atlantic salmon embryos were HS treated during somitogenesis. Ten months later these individuals displayed a high prevalence of caudal vertebral column condensations (27-34%). The defects were located caudally of the abdominal cavity, displaying an even distribution in this region independent of time of HS. To determine if HS disturbed vertebral development during somitogenesis, two genes coding for markers of skeletal development were identified, namely, the secreted protein Shh (Sashh) and the transcription factor Twist (Satwist). These proteins are involved in the proliferation and specification of presumptive skeletal cells (sclerotome) in vertebrates. The spatial expression pattern of sashh and satwist in salmon indicated a functional conservation of these proteins. Furthermore, HS embryos displayed expressional disturbance in both sashh and satwist, indicating an effect of HS on sclerotomal cell patterning. However, the HS-protecting ability in embryos seems to be individually regulated because reduction in gene expression was not detected at all stages; in addition, HS did not induce somitic disturbance and vertebral deformity in all embryos. PMID- 15798922 TI - Meningitis following basal skull fracture in two in-line skaters. AB - INTRODUCTION: In-line skating has been reported to cause severe head injury. Basilar skull fracture (BSF) is associated with a high risk of complication. CASE REPORT: We report two children who had bacterial meningitis following seemingly trivial in-line skating injuries. In both, anterior BSF was diagnosed retrospectively following occurrence of Streptococcus pneumoniae meningitis. DISCUSSION: The clinical signs indicating BSF depend on the fracture location. Plain skull radiography and computed tomography (CT) are not sensitive enough to detect thin fractures in the anterior cranial fossa. We argue that high resolution multiple-plane CT and coronal T2-weighted magnetic resonance imaging are indicated to diagnose BSF. PMID- 15798921 TI - Positron emission tomography for the early postsurgical evaluation of pediatric brain tumors. AB - OBJECT: The object was to study the value of postoperative positron emission tomography (PET) to assess the extension of brain tumor resection. METHODS: Twenty children operated on for total resection of a glial tumor (18 low-grade, 2 anaplastic) presented a signal on postoperative magnetic resonance (MR) images raising the question of a possible tumor residue. PET was performed early ((18)F Fluoro-deoxyglucose in 1, (11)C-methionine in 16, both in 3) to further characterize the nature of the abnormal MR signal in order to consider second look surgery. An increased tracer uptake found in 14 children led to reoperation on 11 of them, confirming the tumor histologically. No (11)C-methionine uptake led to a conservative attitude in 6 children in whom MR imaging follow-up showed no tumor progression. CONCLUSIONS: The early postoperative PET, especially with (11)C-methionine, appears to be a valid basis for complementary therapeutic decisions, especially second-look surgery, in glial tumors for which a radical resection is a key factor for prognosis. PMID- 15798923 TI - [Status and integration of radiology in a large clinical center]. AB - Radiology has undergone enormous technological and hence organizational changes in the last few decades. These changes occurred more or less in unison with total state expenditures for healthcare, whereas individual disposable income increased twice as much in the same timeframe (1988-2003). Costs for medical technology account for approximately 3-5% of national healthcare expenditures. On the national level, but also in a central hospital setting, radiology does not constitute a major cost factor. Tomographic investigations have increased by 2-3% in recent years and currently represent 30% of all radiological procedures. Increasing sophistication of diagnostics and treatment using genetic and molecular techniques, novel information technologies, and a critical evidence based approach to all functional activity will result in a further explosion of knowledge. Radiology must meet these challenges head on by intensifying all forms of training, from basics to continuing professional education. PMID- 15798924 TI - [Strategies of the nasal mucous membranes for defense against infection. Current knowledge of antimicrobial peptides]. AB - Infections of healthy skin or mucous membranes are rare despite constant exposure to microbial colonization. Nonspecific and specific defense mechanisms correspond in the nose: the mucociliary clearance provides very effective mechanical cleaning. If this protection is insufficient, the nasal epithelium comes into close contact with microorganisms. This triggers the recently discovered production of the body's own specific antimicrobial peptides (AP). It seems that it is difficult for microorganisms to form enzyme-induced resistance to AP. This makes AP highly interesting for the development of new antibiotics. The observation that antimicrobial activity can be induced has also led to the hypothesis that natural substances may possibly stimulate endogenous AP production. If this line of defense also fails, the inflammatory cells migrate from the blood into the tissue and, with their capacity for phagocytosis, the enzymatic attack on bacteria can cause a cellular inflammatory reaction in the nasal mucous membrane. PMID- 15798925 TI - RADPED: an approach to teaching communication skills to radiology residents. AB - BACKGROUND: The Accreditation Council for Graduate Medical Education mandates that radiology residency programs teach communication skills to residents. OBJECTIVE: The purpose of this paper is to present a mnemonic, RADPED, that can be used to enhance communication in the radiology setting. It reminds the resident of the salient points to address during an imaging encounter with pediatric patients and their families for the purpose of enhancing communication. MATERIALS AND METHODS: Recent history and research in medical communication are reviewed. Various communication guides used by primary care physicians, such as SEGUE, and the Kalamazoo consensus statement are discussed. This methodology was adapted into a format that could be used to teach communication skills to radiology residents in the context of an imaging encounter. RESULTS: RADPED reminds the resident to establish rapport with the patient, ask questions as to why the patient and family are presenting for the study, discuss the exam, perform the procedure, use exam distractions, and discuss the results with the referring physician and family when appropriate. SUMMARY: This simple memory aid promotes the key points necessary to optimize the radiology resident's encounter with pediatric patients and their families. PMID- 15798926 TI - Popliteal artery entrapment syndrome in a young girl. AB - Popliteal artery entrapment syndrome (PAES) is well-described in adults, but is an uncommon cause of lower-limb pain in children. We present an 11.5-year-old girl with thrombosed aneurysm of the right popliteal artery, subsequently diagnosed with bilateral type I PAES. Multimodality illustration of the radiological findings is presented. PMID- 15798929 TI - Identification of two novel genes for blackleg resistance in Brassica napus. AB - Blackleg, caused by Leptosphaeria maculans, is a major disease of Brassica napus. Two populations of B. napus DH lines, DHP95 and DHP96, with resistance introgressed from B. rapa subsp. sylvestris, were genetically mapped for resistance to blackleg disease with restriction fragment length polymorphism markers. Examination of the DHP95 population indicated that a locus on linkage group N2, named LepR1, was associated with blackleg resistance. In the DHP96 population, a second locus on linkage group N10, designated LepR2, was associated with resistance. We developed BC1 and F2 populations, to study the inheritance of resistance controlled by the genes. Genetic analysis indicated that LepR1 was a dominant nuclear allele, while LepR2 was an incompletely dominant nuclear resistance allele. LepR1 and LepR2 cotyledon resistance was further evaluated by testing 30 isolates from Canada, Australia, Europe, and Mexico. The isolates were from B. napus, B. juncea, and B. oleracea and represented different pathogenicity groups of L. maculans. Results indicated that LepR1 generally conferred a higher level of cotyledon resistance than LepR2. Both genes exhibited race-specific interactions with pathogen isolates; virulence on LepR1 was observed with one isolate, pl87-41, and two isolates, Lifolle 5, and Lifolle 6, were virulent on LepR2. LepR1 prevented hyphal penetration, while LepR2 reduced hyphal growth and inhibited sporulation. Callose deposition was associated with resistance for both loci. PMID- 15798933 TI - Functional neuroimaging of sleep. AB - Sleep and sleep disorders have traditionally been viewed from a polysomnographic perspective. Although these methods provide information on the timing of various stages of sleep and wakefulness, they do not provide information regarding function in brain structures that have been implicated in the generation of sleep and that may be abnormal in different sleep disorders. Functional neuroimaging methods provide information regarding changes in brain function across the sleep wake cycle that provides information for models of sleep dysregulation in a variety of sleep disorders. Early studies show reliable increases in function in limbic and anterior paralimbic cortex in rapid eye movement (REM) sleep and decreases in function in higher-order cortical regions in known thalamocortical networks during non-REM sleep. Although most of the early work in this area has been devoted to the study of normal sleep mechanisms, a collection of studies in diverse sleep disorders such as sleep deprivation, depression, insomnia, dyssomnias, narcolepsy, and sleep apnea suggest that functional neuroimaging methods have the potential to clarify the pathophysiology of sleep disorders and to guide treatment strategies. PMID- 15798934 TI - Sleep and stroke. AB - More than 50% of stroke patients have sleep-disordered breathing (SDB), mostly in the form of obstructive sleep apnea (OSA). SDB represents both a risk factor and a consequence of stroke. The presence of SDB has been linked with poorer long term outcome and increased long-term stroke mortality. Continuous positive airway presure is the treatment of choice for OSA. Oxygen and other forms of ventilation may be helpful in other (e.g., central) forms of SDB. SDB can improve spontaneously after stroke. About 20 to 40% of stroke patients have sleep-wake disorders (SWD), mostly in form of insomnia, excessive daytime sleepiness/fatigue, or hypersomnia (increased sleep needs). Depression, anxiety, SDB, stroke complications, and medications may contribute to SWD and should be addressed first therapeutically. Brain damage per se, often at thalamic or brainstem level, can be also a cause of persisting SWD. In these patients, hypnotics, dopeminergic agents, and stimulants (e.g., modafinil) can be attempted. PMID- 15798935 TI - Sleep disorders and neuromuscular disease. AB - Breathing is the most important executive function of sleep. Compromise of the ventilatory mechanism is the principal consequence of a neuromuscular dysfunction in the individual who is asleep. This includes alterations of the lower motor neuron, the neuromuscular junction, and muscle. The primordial phenomenon in rapid eye movement (REM) sleep respiratory physiology is that the diaphragm assumes all functions of respiration. Any dysfunction of the diaphragm, whether neurogenic or neuromuscular, will interfere with breathing during REM sleep. So prevalent are sleep respiratory difficulties in patients with neuromuscular disorders that there should be a low threshold to obtain nocturnal polysomnography in these patients with sleep complaints. In patients with a neuromuscular disorder and nocturnal ventilatory compromise, positive airway pressure ventilation improves the quality of sleep and in doing so improves the overall quality of life. PMID- 15798936 TI - Sleep and neurodegenerative diseases. AB - Sleep disturbances are common in neurodegenerative diseases. Disturbed sleep can result in fatigue, irritability, morning headaches, impaired motor and cognitive skills, depression, and daytime somnolence. The major sleep complaints include: insomnia, hypersomnia, parasomnia, excessive nocturnal motor activity, circadian sleep-wake rhythm disturbance, and respiratory dysrhythmia. The pathogenetic mechanisms of sleep disturbances may be secondary to direct structural alteration of the sleep-wake generating neurons or from several other indirect mechanisms. At the biochemical level, neurodegenerative diseases may be largely classified as tauopathies, alpha-synucleinopathies, and other diseases. Overnight polysomnography (PSG) and multiple sleep latency test are the two most important diagnostic laboratory tests in the evaluation of sleep disturbances. Management of sleep disturbances is complex and is based primarily on the nature of the sleep disturbance. The clinical profiles, pathogenetic mechanisms, PSG findings, and management issues are discussed here with reference to some common neurodegenerative diseases. PMID- 15798937 TI - Sleep in the geriatric patient population. AB - Older patients often suffer from sleep disturbances caused by age-related physiological changes, polypharmacy, changes in circadian rhythm, retirement, and loss of spouse. Many seniors have readily diagnosed sleep disorders that can be treated to achieve improvement in daytime somnolence or cognitive impairments. After a discussion of normal age-related changes in sleep, specific sleep pathologies common in the elderly will be reviewed, including sleep apnea, restless legs syndrome, advanced sleep phase syndrome, and rapid eye movement sleep behavior disorder. PMID- 15798938 TI - Sleep disorders in multiple sclerosis. AB - Sleep disorders are pervasive in patients with multiple sclerosis (MS) although clinically underrecognized by most physicians. The most common sleep disorders seen in patients with MS include insomnia, nocturnal movement disorders, sleep disordered breathing, narcolepsy, and rapid eye movement sleep behavior disorder. Factors that influence the quality of sleep in this patient population include pain, nocturia, depression, medication effect, location of lesions, and disease severity. Disrupted sleep has the potential to cause daytime somnolence, increased fatigue, and nonrefreshing sleep, and it may be associated with dangerous respiratory events. Awareness and treatment of these conditions is vital to improving health and quality of life in patients with MS. PMID- 15798939 TI - Sleep-related headache syndromes. AB - An association between headache and sleep is long recognized in the medical literature and deemed highly clinically relevant for at least a portion of headache sufferers. Sleep-related headaches are not well understood, but recent advances in the neurophysiology of sleep and headache shed light on common biological processes potentiating their association. Respective diagnostic nosologies for headache and sleep disorders offer limited guidance but are evolving to better represent the known associations. A rational approach based on the available empiricism can assist the clinician in the assessment and treatment of headache. Each headache evaluation should include at least a brief sleep history. Headaches that occur during or after sleep, so called morning headache, may be secondary to a sleep disorder and call for a specific treatment. This case is best established for sleep-related breathing disorders, and treatment of apnea often leads to resolution of the headache. Although pure sleep-related headaches, such as hypoxemia-related headache and hypnic headache, are less prevalent and more easily recognized, primary headaches are often impacted to some degree by sleep. Additionally, common headache medications and comorbid conditions alter sleep architecture. Management of primary headache may be facilitated by attention to sleep complaints. PMID- 15798940 TI - Insomnia in neurological diseases. AB - Insomnia is the most common sleep complaint. Insomnia is not a disease itself but mostly a clinical sign of an underlying disease. Degenerative and vascular diseases involving the central nervous system (CNS) may impair sleep either as a result of the brain lesion or because of illness-related discomfort (motor immobility, social and familial impairment, depression, drugs). Some neurological conditions characterized by movement disorders that start or persist during sleep hinder sleep onset and/or sleep continuity, causing a poor sleep complaint. CNS lesions and/or dysfunction in three specific neurological conditions (fatal familial insomnia, Morvan's chorea, and delirium tremens) impair the basic mechanisms of sleep generation inducing a syndrome in which the inability to sleep is consistently associated with motor and sympathergic overactivation. Agrypnia excitata is the term that aptly defines this generalized overactivation syndrome. PMID- 15798941 TI - Psychiatric disorders associated with disturbed sleep. AB - Disturbed sleep is a feature of many psychiatric disorders. When clinicians are investigating insomnia, excessive daytime sleepiness, as well as parasomnia, they need to consider whether these complaints might be secondary to a psychiatric process. The association that is best supported by scientific evidence is between insomnia and mood disorders. Nonetheless, other psychiatric conditions including anxiety, psychosis, chemical dependency, and attention deficit hyperactivity disorder are closely linked with alteration in sleep quality and quantity. Treatment plans should take into account the nature of sleep symptomatology as well as the effect of medications on sleep. Ideally, management should include a combination of medications and behavioral treatment. PMID- 15798942 TI - Physiology and psychology of dreams. AB - The discovery of the close association between rapid eye movement (REM) sleep and dreaming and development of sleep laboratory techniques ushered in a new era in the study of dreams. For the first time, direct and systematic investigation could be made of such topics as the occurrence, qualities, recollection, and childhood development of dreaming. Experimental methodologies permitted investigation of the responsiveness of dreams to external stimulation and the effects of deprivation of REM sleep. Much effort was devoted to searching for parallels between physiological aspects of REM sleep and characteristics of associated dreams, with modest results. The leading theory of dreaming in the early decades of this research was the psychoanalytic, which views dreams as highly meaningful reflections of unconscious mental functioning. With developments in understanding of the neurophysiology of REM sleep, new theories of dreaming were proposed. The most prominent, the activation-synthesis hypothesis, derived its view of dreaming directly from the neurophysiology of REM sleep, in particular the role of the brain stem, and in its original form regarded dreams as not essentially meaningful. Further developments in neurobiological research, including lesion and brain imaging studies, have established a clearer view of the functional neuroanatomy of REM sleep and dreaming. To what degree, and in what way, implications can be drawn from these findings for the psychology of dreaming is controversial. Some more recent theories of dreaming emphasize an adaptive function related to emotion and a role in learning and memory consolidation. PMID- 15798943 TI - Sleep and pain: interaction of two vital functions. AB - Disturbed sleep is a key complaint of people experiencing acute and chronic pain. These two vital functions, sleep and pain, interact in complex ways that ultimately impact the biological and behavioral capacity of the individual. Polysomnographic studies of patients experiencing acute pain during postoperative recovery show shortened and fragmented sleep with reduced amounts of slow wave and rapid eye movement (REM) sleep, and the recovery is accompanied by normalization of sleep. Objective assessments of sleep in patients with various chronic pain conditions have been less definitive with some studies showing fragmented and shortened sleep and others showing normal sleep. Although daytime fatigue is a frequent complaint associated with complaints of pain-related disturbed sleep, objective assessments of daytime sleepiness reveal minimally elevated levels of sleepiness and emphasize the importance of distinguishing sleepiness and fatigue. The pain-sleep nexus has been modeled in healthy pain free subjects and the studies have demonstrated the bidirectionality of the sleep pain relation. Given this bidirectionality, treatment must focus on alleviation of both the pain and sleep disturbance. Few of the treatment studies have done such, and as a result no clear consensus on treatment approaches, much less on differential etiology-based treatment strategies, has emerged. PMID- 15798944 TI - Neurocognitive consequences of sleep deprivation. AB - Deficits in daytime performance due to sleep loss are experienced universally and associated with a significant social, financial, and human cost. Microsleeps, sleep attacks, and lapses in cognition increase with sleep loss as a function of state instability. Sleep deprivation studies repeatedly show a variable (negative) impact on mood, cognitive performance, and motor function due to an increasing sleep propensity and destabilization of the wake state. Specific neurocognitive domains including executive attention, working memory, and divergent higher cognitive functions are particularly vulnerable to sleep loss. In humans, functional metabolic and neurophysiological studies demonstrate that neural systems involved in executive function (i.e., prefrontal cortex) are more susceptible to sleep deprivation in some individuals than others. Recent chronic partial sleep deprivation experiments, which more closely replicate sleep loss in society, demonstrate that profound neurocognitive deficits accumulate over time in the face of subjective adaptation to the sensation of sleepiness. Sleep deprivation associated with disease-related sleep fragmentation (i.e., sleep apnea and restless legs syndrome) also results in neurocognitive performance decrements similar to those seen in sleep restriction studies. Performance deficits associated with sleep disorders are often viewed as a simple function of disease severity; however, recent experiments suggest that individual vulnerability to sleep loss may play a more critical role than previously thought. PMID- 15798945 TI - Case studies in sleep medicine. PMID- 15798947 TI - The effect of a high-carbohydrate meal on postprandial thermogenesis and sympathetic nervous system activity in boys with a recent onset of obesity. AB - The purpose of the present study was to investigate the thermic effect of food (TEF) and sympathetic nervous system (SNS) activity in obese boys. Ten obese (9.2+/-0.4 years) and 13 lean boys (8.8+/-0.4 years) were examined for energy expenditure and fat oxidation measured via indirect calorimetry for 3 hours after a high-carbohydrate (HC; 70% carbohydrate, 20% fat, and 10% protein) or a high fat (HF; 20% carbohydrate, 70% fat, and 10% protein) meal served on 2 different days at random. The activity of the SNS was assessed by means of a power spectral analysis of the heart rate variability. The TEF, expressed as a percentage of the consumed energy, was significantly lower in obese boys than in lean boys after the HC meal; however, such a difference was not observed after the HF meal. Multiple regression analysis revealed that obesity was a significant variable contributing to the variances in the TEF induced by the HC meal. Moreover, after the HC meal, the boys with a recent onset of obesity (duration, <3 years) manifested a lower TEF as well as a reduced very low frequency component of the heart rate variability, an index of thermoregulatory SNS functions, compared with the remaining obese and lean boys. In conclusion, obese boys possessed normal metabolic and sympathetic responses to the HF meal but showed a diminished thermogenic response to the HC meal, especially during the early phase of obesity. PMID- 15798946 TI - Phenotype-based treatment of dietary obesity: differential effects of fenofibrate in obesity-prone and obesity-resistant rats. AB - High-fat diets (HFDs) promote hyperphagia and adiposity in animals and human beings. To test the hypothesis that limitations on fat oxidation underlie this propensity for diet-induced obesity, rats were treated with fenofibrate, which enhances fat oxidation mainly in liver by inducing expression of enzymes and proliferation of organelles involved in fatty acid oxidation. Male Sprague-Dawley rats were fed a HFD (42% fat calorie) for 2 weeks. Rats ranked in the top and bottom thirds for weight gain during this feeding period were designated as obesity prone (OP) and obesity resistant (OR), respectively. Fenofibrate was added to the HFD (0.025% wt/wt) for half of the OP and OR rats. During the next 10 days, fenofibrate treatment significantly (P<.05) reduced food intake, weight gain, feed efficiency, and adiposity in OP rats to levels seen in control OR rats, but had no such effects in OR rats. Fenofibrate treatment increased whole body fatty acid oxidation, and in liver, the expression of carnitine palmitoyl transferase I only in OP rats, but enhanced expression of acyl-CoA oxidase in both OP and OR rats. Restricting food intake of OP rats to levels seen in rats given fenofibrate similarly reduced weight gain but had little effect on weight of fat pads. Treatment with the daily dosage of fenofibrate given as a bolus did not produce a conditioned flavor aversion. These results suggest that enhancement of mitochondrial fatty acid oxidation in liver may be an effective phenotype based treatment strategy for dietary obesity. PMID- 15798948 TI - The reciprocal association of adipocytokines with insulin resistance and C reactive protein in clinically healthy men. AB - In experimental models, adiponectin improves and tumor necrosis factor alpha (TNF alpha ) impairs insulin action, and the expression of these adipocytokines seems to have a reciprocal regulation. The aim was to examine whether in a cross sectional study, associations supporting this concept may be found in 58-year-old clinically healthy men, and also the relation to C-reactive protein (CRP). In 102 men, euglycemic hyperinsulinemic clamp was used to assess glucose infusion rate (GIR). Total body fat (dual-energy x-ray absorptiometry), plasma adiponectin (radioimmunoassay), TNF-alpha , and CRP (enzyme-linked immunosorbent assay) were measured. Adiponectin correlated positively to GIR (r=0.33, P<.001) and negatively to total fat mass (r=-0.29, P=.004), whereas TNF- alpha showed reverse associations (r=-0.31, P<.01, and r=0.31, P<.01). Adiponectin and TNF- alpha were negatively correlated (-0.28, P=.006). An interaction term (TNF- alpha /adiponectin ratio) and body fat together explained 31.3% (P<.001) in GIR variability. The odds ratio for having insulin resistance was 9.3 (95% CI, 2.2 38.9) in subjects with TNF-alpha values above and adiponectin levels below the median, as compared to subjects with TNF- alpha values below and adiponectin levels above the median. Total fat and TNF-alpha , but not adiponectin, were significantly associated with log CRP (R2=20%, P<.001). In conclusion, this study in man showed that plasma adiponectin and TNF-alpha were independently and reversely associated with insulin resistance. C-reactive protein levels were related to TNF-alpha and obesity. PMID- 15798949 TI - Different tumor necrosis factor-alpha-associated leptin expression in rats with dimethylnitrosamine and bile duct ligation-induced liver cirrhosis. AB - Although serum leptin concentrations are reported by several studies to increase in patients with liver cirrhosis, the mechanisms underpinning this increase remain unclear. Circulating tumor necrosis factor alpha (TNF-alpha) concentrations are also recognized to increase in liver cirrhosis. Furthermore, TNF-alpha administration to rodents results in increased expression and secretion of leptin from adipose tissue in a manner dependent on type 1 TNF-alpha receptor (TNF-RI). The present study was undertaken to examine adipose leptin expression and to explore potential relationships between leptin expression and TNF-alpha in subjects with liver cirrhosis. Liver cirrhosis was induced in male Sprague-Dawley rats by dimethylnitrosamine (DMN) administration or by common bile duct ligation (BDL). Ad libitum and pair-fed animals constituted controls. Serum leptin and TNF alpha concentrations were determined by immunoassay. Gene expression was determined by the reverse transcription-polymerase chain reaction, and protein levels were measured by Western blotting. Serum leptin values after adjustment of body fat mass in DMN-treated rats were significantly higher than in pair-fed or ad libitum groups. Leptin mRNA and protein levels in epididymal fat in DMN rats increased by 1.8-fold and 2.3-fold, respectively, as compared with ad libitum controls, and by 4-fold and 6-fold, respectively, as compared with the pair-fed group. Epididymal TNF-alpha and membranous TNF-RI (mTNF-RI) concentrations were both 2.3 times higher in DMN rats than in ad libitum controls but did not differ between ad libitum and pair-fed groups. Adipose leptin protein levels correlated directly with TNF-alpha and mTNF-RI concentrations in combined DMN, ad libitum, and pair-fed rats (r=0.64 and r=0.49, respectively; P<.05). In BDL-treated rats, however, serum and adipose leptin concentrations were identical to those in ad libitum controls despite 2.1-fold and 2.4-fold increase in epididymal TNF-alpha and mTNF-RI, respectively. TNF-alpha administration to fasting control animals increased serum and adipose leptin concentrations significantly. The observed TNF alpha-associated leptin up-regulation in DMN-induced, but not in BDL-induced, cirrhotic rats is consistent with distinctly different roles for TNF-alpha in rats with nonbiliary, as opposed to biliary, cirrhosis. PMID- 15798950 TI - Iron loading increases cholesterol accumulation and macrophage scavenger receptor I expression in THP-1 mononuclear phagocytes. AB - Epidemiological studies have established that a high level of iron body stores is associated with increased risk of acute coronary heart disease. To explain this association, it has been proposed that iron catalyzes the production of highly reactive forms of free oxygen species, and thus, promotes low-density lipoprotein (LDL) oxidation, a lipoprotein that plays a critical role in atherogenesis. However, few studies have provided evidence to support this hypothesis. In the present study, we determined the effect of iron loading of THP-1 mononuclear phagocytes on LDL metabolism. We demonstrated that iron loading of THP-1 cells stimulated conjugated diene formation in LDL in the culture medium. In addition, iron loading of THP-1 cells significantly increased cholesteryl ester accumulation in cells exposed to native LDL, suggesting that during the incubation of the cells with native LDL, the LDL became oxidized and was taken up by the cells. We further demonstrated that the degradation of 125I-oxidized LDL was significantly increased in iron-loaded THP-1 cells. Lastly, we demonstrated that iron loading of THP-1 cells stimulated scavenger receptor expression in these cells. In conclusion, this study demonstrates that loading of mononuclear phagocytes with iron leads to oxidization of LDL, increased cellular cholesterol accumulation and scavenger receptor expression, and supports the hypothesis that increased macrophage iron levels promote atherogenesis. PMID- 15798951 TI - Methimazole-induced hypothyroidism paradoxically decreases homocysteine. AB - Clinical hypothyroidism is associated with hyperhomocysteinemia, whereas the opposite is seen in hyperthyroidism. The effects of mild thyroid dysfunction on homocysteine concentrations are not known. We performed the following study to investigate this. Total homocysteine, vitamins B6 and B12, folate, fibrinogen, plasminogen activator inhibitor type 1, and lipids were measured in 11 subjects at baseline and after methionine loading. Subjects began methimazole (MMI), 40 mg daily, and were restudied during 2 stages of hypothyroidism. Liothyronine was added and subjects were restudied once thyrotropin normalized. Methimazole was stopped and studies were repeated during 2 stages of hyperthyroidism. Data were analyzed using repeated-measures analysis of variance. Post-methionine homocysteine decreased in each hypothyroid study compared with baseline (28.8+/ 10.7, 27.5+/-9.9 vs 34.4+/-9.2 micromol/L, respectively). In addition, both fasting and post-methionine homocysteine decreased in the euthyroid/MMI study arm compared with baseline despite equivalent thyrotropin values (fasting, 7.5+/-3.0 vs 8.8+/-3.5 micromol/L, P<.05; and post-methionine, 27.2+/-10.6 vs 34.4+/-9.2 micromol/L, P<.05, respectively). Fasting homocysteine decreased in the first hyperthyroidism study arm compared with baseline (6.6+/-2.3 vs 8.8+/-3.5 micromol/L, P<.05) and post-methionine homocysteine decreased in both hyperthyroid arms compared with baseline (25.2+/-8.1, 24.2+/-10 vs 34.4+/-9.2 micromol/L, P<.05 respectively). In conclusion, mild thyroid dysfunction changes homocysteine metabolism. Unexpectedly, our results suggest a homocysteine lowering effect of MMI. PMID- 15798952 TI - Associations of middle-aged mother's but not father's body mass index with 18 year-old son's waist circumferences, birth weight, and serum hepatic enzyme levels. AB - Mitochondrial dysfunction has been reported to contribute to insulin resistance (IR) in the elderly and type 2 diabetes. To test this hypothesis, we examined relations of insulin resistance in young men to their mother's body mass index (BMI) and compared with those to their father's BMI, because as a rule, mitochondrial DNA is exclusively maternally inherited and because mitochondria are fundamental in mediating effects on energy dissipation. We measured heights, weights, waist circumference, systolic and diastolic blood pressure (BP), and biochemical variables in sera from 193 male college students aged 18 to 20 years after an overnight fast. Birth weight was available from 184 students. Self reported heights and weights of their parents were obtained from 148 students. Insulin resistance and insulin secretion were estimated using homeostasis model assessment (HOMA-IR and HOMA-beta, respectively). Mother's BMI was associated with their son's birth weight (r=0.23, P=.008), BMI (r=0.37, P<.0001), waist circumference (r=0.42, P<.0001), fasting insulin (r=0.19, P=.02), and HOMA-IR (r=0.18, P=.03) but not with fasting glucose, HOMA-beta , and systolic and diastolic BP. In addition, high-density lipoprotein cholesterol and lipoprotein(a) [Lp(a)] were inversely associated with mother's BMI (r=-0.21, P=.01 and r=-0.17, P=.03, respectively). Furthermore, there were significant associations with aspartate (r=0.20, P=.01) and alanine (r=0.28, P=.0008) aminotransferase and gamma-glutamyl transpeptidase (r=0.30, P=.0003), all of which are associated with mitochondrial function. In contrast, none of those variables were associated with father's BMI, except for Lp(a), which showed a significant and inverse association (r=-0.17, P=.05). After adjustment for sons' BMI, waist circumference and 3 hepatic enzymes were associated with mother's BMI, whereas Lp(a) was associated with both mother's and father's BMI. In multiple regression analysis for HOMA-IR as a dependent variable, BMI of their own (beta=.10, P<.0001) and of their mothers (beta=.04, P=10) and birth weight (beta= .27, P=.10) emerged as determinants of HOMA-IR of the students(R2=0.30). Our results are consistent with clinical observations of a greater risk of transmission of type 2 diabetes from the mother than the father and suggest that son's IR may be influenced by maternal effect as well as their adiposity. PMID- 15798953 TI - Serum resistin level among healthy subjects: relationship to anthropometric and metabolic parameters. AB - Resistin is a novel adipocyte-secreted hormone that has been proposed to be the link between obesity and diabetes, although little appears to be known regarding the physiological role of resistin in human beings. We aimed to explore the relationship between serum resistin level and certain anthropometric and metabolic parameters. Seventy-one healthy subjects with a mean body mass index of 23 kg/m 2 or greater were recruited in this study. Anthropometric measurements including height, weight, body mass index, waist and hip circumferences, waist-to hip ratio, and blood pressure were recorded. Insulin resistance was measured by homeostasis model assessment (HOMA). Fasting serum resistin, insulin and plasma glucose, lipid profiles, and uric acid levels were measured. The results revealed that serum resistin level did not correlate with any markers for adiposity, blood pressure, fasting plasma glucose, or uric acid level for either sex. Serum resistin level correlated negatively with fasting insulin level (gamma=-0.455, P=.006) and HOMA (gamma=-0.455, P=.006) in women but not in men. Serum resistin level only correlated negatively with high-density lipoprotein cholesterol (HDL C) level in men (gamma=-0.347, P=.038); there was no correlation between serum resistin level and lipid profiles in women. Multiple linear regression analysis using the logarithm of resistin as a dependent variable revealed that only HDL-C level (beta=-.058, P=.019) was an independent significant predictor for resistin in men; however, the analysis revealed that HDL-C level (beta=-.044, P=.029) and HOMA (beta=-.719, P=.004) were independent significant predictors for resistin in women. In conclusion, resistin is not related to adiposity, blood pressure, insulin resistance, fasting plasma glucose level, and most lipid profiles. Resistin correlates negatively with HDL-C level for both sexes. The role of resistin in metabolic syndrome warrants further investigation. PMID- 15798954 TI - Association of low adiponectin levels with the metabolic syndrome--the Chennai Urban Rural Epidemiology Study (CURES-4). AB - The aim of the study was to assess the relation of adiponectin levels with the metabolic syndrome in Asian Indians, a high-risk group for diabetes and premature coronary artery disease. The study was conducted on 100 (50 men and 50 women) type 2 diabetic subjects and 100 age and sex matched subjects with normal glucose tolerance selected from the Chennai Urban Rural Epidemiology Study, an ongoing population study in Chennai in southern India. Metabolic syndrome was defined using modified Adult Treatment Panel III (ATPIII) guidelines. Adiponectin values were significantly lower in diabetic subjects (men: 5.2 vs 8.3 microg/mL, P=.00l; women: 7.6 vs 11.1 microg/mL, P<.00l) and those with the metabolic syndrome (men: 5.0 vs 6.8 microg/mL, P=.01; women: 6.5 vs 9.9 microg/mL, P=.001) compared with those without. Linear regression analysis revealed adiponectin to be associated with body mass index (P<.05), waist circumference (P<.01), fasting plasma glucose (P=.001), glycated hemoglobin (P<.001), triglycerides (P<.00l), high-density lipoprotein (HDL) cholesterol (P<.001), cholesterol/HDL ratio (P<.00l), and insulin resistance measured by homeostasis assessment model (P<.00l). Factor analysis identified 2 factors: factor 1, negatively loaded with adiponectin and HDL cholesterol and positively loaded with triglycerides, waist circumference, and insulin resistance measured by homeostasis assessment model; and factor 2, with a positive loading of waist circumference and systolic and diastolic blood pressure. Logistic regression analysis revealed adiponectin to be negatively associated with metabolic syndrome (odds ratio [OR], 0.365; P<.001) even after adjusting for age (OR, 0.344; P<.00l), sex (OR, 0.293; P<.001), and body mass index (OR, 0.292; P<.00l). Lower adiponectin levels are associated with the metabolic syndrome per se and several of its components, particularly, diabetes, insulin resistance, and dyslipidemia in this urban south Indian population. PMID- 15798955 TI - Hypoadiponectinemia is caused by chronic blockade of nitric oxide synthesis in rats. AB - Adiponectin is an adipocyte-derived anti-atherogenic protein. Adiponectin levels are decreased in patients and animal models with obesity, diabetes, and coronary artery disease. However, the mechanism by which adiponectin levels are reduced remains unknown. Since hypoadiponectinemia is closely linked to endothelial dysfunction, we examined the regulation of adiponectin in a rat model of chronic blockade of nitric oxide (NO) synthesis by N omega-nitro-L-arginine methyl ester (L-NAME). Decreased production of NO and increased production of O2- were observed in aorta from L-NAME-treated rats. Plasma adiponectin levels and adiponectin mRNA levels of adipose tissue were markedly decreased in L-NAME treated rats. Cotreatment of pioglitazone (PIO) or allopurinol (ALL) with L-NAME restored plasma adiponectin concentration and fat adiponectin mRNA levels to control levels. Thus, adiponectin levels were decreased in L-NAME-treated rats, however, they returned to normal following administration of PIO due to transcriptional activation of the adiponectin gene, as well as administration of ALL, likely due to elimination of oxidative stress. Oxidative stress appears to be an important cause of hypoadiponectinemia. PMID- 15798956 TI - Association study of G1704T and G82S polymorphisms of RAGE gene for microalbuminuria in Japanese type 2 diabetic patients. AB - To clarify whether polymorphisms G1704T and G82S of the RAGE gene were related to microalbuminuria, we performed a case-control study in Japanese type 2 diabetic patients. Polymorphisms G1704T and G82S of the RAGE gene were examined with genomic DNA obtained from 116 type 2 diabetic patients with microalbuminuria (urinary albumin/creatinine ratio between 30 and 300 mg/g of creatinine) (microalbuminuria group), and 232 patients with normoalbuminuria (urinary albumin/creatinine ratio <30 mg/g of creatinine) (normoalbuminuria group). The genotype distribution and T allele frequency of G1704T (9.9%) and S allele frequency of G82S (14.2%) in the microalbuminuria group did not significantly differ from those (T allele frequency, 8.4%; S allele frequency, 12.3%) in the normoalbuminuria group. There were no differences among the genotypes of G1704T and G82S of the RAGE gene regarding age, duration of diabetes, body mass index, glycosylated hemoglobin (HbA1c), blood pressure, and serum lipid levels. These data suggest that G1704T and G82S polymorphisms of the RAGE gene are not related to microalbuminuria in Japanese type 2 diabetic patients. PMID- 15798957 TI - Influence of sex and beta2 adrenergic receptor haplotype on resting and terbutaline-stimulated whole body lipolysis. AB - beta 2 adrenergic receptors ( beta 2 ARs) are important mediators of lipolysis. The beta 2 AR gene is highly polymorphic. To determine the contribution of beta 2 AR polymorphisms to variability in whole body lipolysis, we compared basal and terbutaline-stimulated lipolytic rates (Ra) using tracer techniques in 14 healthy, non-obese males (n=7) and females (n=7) who were homozygous for Cys 19/Arg16/Gln27 or Arg-19/Gly16/Glu27 haplotypes. Fasting (overnight) Ra values were higher in females compared to males. Mean+/-SD Ra, Ra/body weight, Ra/fat free mass, Ra/fat, and Ra/energy expenditure rates in males and females were 155+/-46 vs 311+/-111 micromol/min (P=.007); 2.0+/-0.61 vs 5.2+/-2.3 micromol/(min kg) (P=.006); 2.5+/-0.75 vs 7.8+/-3.4 micromol/(min kg) (P=.003); 10+/-3.7 vs 17+/-7.4 micromol/(min kg) (P=.09); and 144+/-45.5 vs 392+/-111 micromol/d (P=.0001), respectively. Mean+/-SD basal glycerol concentrations were higher in females compared to males: 62+/-5.6 vs 36+/-17 micromol/L (P=.003). Basal glycerol concentrations and Ra values were similar by beta2 AR haplotype. Basal glucose and insulin concentrations tended to be higher in males compared to females and were similar by haplotype. Terbutaline-stimulated changes in glycerol concentrations were variable and are not related to either sex or haplotype. We conclude that compared to haplotype, sex is a more important determinant of basal lipolysis after a 12-hour fast in healthy, non-obese individuals. PMID- 15798958 TI - Altered body composition and metabolism in the male offspring of high fat-fed rats. AB - An intrauterine environment may play a role in predisposing a developing fetus to metabolic diseases during adulthood. We investigated the hypothesis that a maternal diet high in omega-6 polyunsaturated fat can modify the programming of an offspring's glucose tolerance, insulin sensitivity, body composition, lipid metabolism, and insulin signaling. High omega-6 polyunsaturated fat diets were fed to female rats 4 weeks before mating and throughout the gestation period. The offspring were maintained on chow diet. At 3 months of age, indirect calorimetry, oral glucose tolerance tests, and dual x-ray absorptiometry measurements were performed. Triglyceride content and beta-hydroxyacyl coenzyme A dehydrogenase activity were determined in the liver and quadriceps muscle. Expression levels of key insulin signaling pathway proteins were measured in the liver and quadriceps muscle of the 3-month-old offspring. Offspring from the fat-fed dams had significantly increased proportions of both total body fat and abdominal fat. All offspring displayed normal insulin sensitivity and glucose tolerance, although the offspring from the fat-fed dams were significantly more hyperinsulinemic 15 minutes after an oral glucose challenge. Whole body fuel oxidation was not altered. The offspring of fat-fed dams had significantly elevated liver triglyceride content. Insulin signaling protein expression levels in the offspring of fat-fed dams were consistent with reduced hepatic insulin sensitivity but increased quadriceps insulin sensitivity. A maternal diet high in omega-6 polyunsaturated fat evokes programming within the metabolic processes of the offspring that may predispose the offspring to the development of metabolic diseases. PMID- 15798959 TI - Very long chain fatty acids (policosanols) and phytosterols affect plasma lipid levels and cholesterol biosynthesis in hamsters. AB - The aim of the current study was to examine the effects of very long chain fatty acids (VLCFA) alone at 2 dietary levels, or in combination of VLCFA at the lower level with lecithin (LT) or phytosterols (PS), on lipid profiles and cholesterol biosynthesis in hamsters. Seventy-five male Golden Syrian hamsters, weighing 100 to 120 g, were fed a regular rodent chow for 2 weeks before being randomly assigned into 5 groups of 15 animals each fed semisynthetic diets for 4 weeks. Group 1 was given a control diet that contained 0.25% cholesterol and 5% fat with a polyunsaturated to saturated fatty acids ratio of 0.4. Groups 2 to 5 were fed the control diet and given 25 mg/kg BW per day of VLCFA (Licowax) (VLCFA25), 50 mg/kg BW per day of VLCFA (VLCFA50), 25 mg/kg BW per day of VLCFA+1000 mg/kg BW per day of LT (VLCFA25/LT), and 25 mg/kg BW per day of VLCFA+1000 mg/kg BW per day of PS (Cholestatin, VLCFA25/PS), respectively. Results showed that HDL cholesterol (HDL-C) levels were not changed by VLCFA25, although increased by VLCFA50 (P<.05) relative to control. Total cholesterol (T-C) and non-HDL-C levels were not affected by VLCFA25 and VLCFA50 as compared with control. VLCFA25/LT had higher (P<.02) T-C and HDL-C levels than any other treatments and increased (P<.05) liver weight relative to control. In contrast, VLCFA25/PS reduced T-C (P=.0004) and non-HDL-C (P=.007) without effect on HDL-C levels compared with control. Triglyceride levels were not affected by any treatment. Cholesterol biosynthesis rate was higher (P<.05) in animals fed VLCFA25 and VLCFA50 than those fed control or VLCFA25/LT or VLCFA25/PS. Results suggest that PSs can decrease total and non-HDL-C cholesterol, whereas VLCFA may increase HDL-C in hamsters. PMID- 15798960 TI - Metabolism of amino acids by cultured rat Sertoli cells. AB - Sertoli cells support spermatogenesis both spatially and energetically; for this reason, these cells have important adaptations. The energetic metabolism of Sertoli cells was adapted to provide lactate and pyruvate to developing germ cells, because these substrates are essential for spermatocytes and spermatids. In this study, we investigated whether Sertoli cells use alanine, leucine, valine, and glycine as energetic substrates and whether the simultaneous addition of other nutrients, such as glucose and glutamine, might affect the metabolism of these amino acids. Alanine, leucine, valine, and glutamine are almost totally oxidized to CO2 by these cells. In contrast, glycine has been demonstrated to be a poor energetic substrate, being mainly incorporated into proteins, and their metabolism did not change in the presence of palmitic acid, glucose, and/or glutamine. The metabolism of the 3 other amino acids was modified by palmitic acid; besides, glucose changed alanine, leucine, and valine oxidation. Glutamine decreased the oxidation of alanine, leucine, and valine to CO2. Conversely, both alanine and leucine decreased the oxidation of glutamine. Our present findings show that Sertoli cells can adapt its energy metabolism to the oxidative substrates available to fulfill their role in spermatogenic energetic supply. PMID- 15798961 TI - Age and muscle-type modulated role of intramyocellular lipids in the progression of insulin resistance in nondiabetic Zucker rats. AB - The effect of muscle fiber type and maturation on intramyocellular lipid (IMCL) content and its relationship to insulin resistance was investigated. Intramyocellular lipid content in slow-twitch (soleus) and fast-twitch (tibialis anterior, TA) muscles of fa/fa (Zucker fatty rat, ZFR) and age-matched lean (Zucker lean rat, ZLR) Zucker rats were repeatedly measured over 3 months. Intramyocellular lipid levels in both the soleus and the TA were significantly higher in the ZFR relative to the ZLR. For the ZFR, IMCL TA increased by approximately 2-fold from 5.3 to 8.4 weeks of age. No subsequent accumulation of IMCL TA occurred in ZFR from 8.4 up to 13.1 weeks of age. For ZLR, IMCL TA contents steadily decreased from 6.6 to 13.1 weeks of age (-77%, P<.05). In contrast, IMCL levels in the soleus were not significantly altered in either rat strain over the course of the study. Maximum impairment in whole-body insulin sensitivity in ZFR was observed at 9-weeks of age, concomitant with peak IMCL TA accumulation. Insulin-stimulated 2-deoxy-D-glucose (2DG) transport in the TA muscle of 10.2- and 14.1-week-old ZFR was significantly impaired relative to age matched ZLR. Insulin-stimulated glucose uptake in the soleus of ZFR and ZLR decreased (P<.05) as the animals matured (ZFR, -49%; ZLR, -69%). Overall, these results support the hypothesis that fast-twitch glycolytic muscles play a major role during the onset of insulin resistance. In addition, proper timing may govern the success of a pharmacological studies aimed at measuring the impact of insulin-sensitizing drugs on IMCL. PMID- 15798962 TI - Improvement of liver function parameters in patients with type 2 diabetes treated with thiazolidinediones. AB - To increase our understanding of the effect of thiazolidinediones, a new class of antidiabetic drugs, on liver function as well as glycemic control, we investigated liver function before, during, and after treatment with troglitazone and pioglitazone. A total of 32 patients with type 2 diabetes were studied. Glycemic control and liver function were measured before, during, and after 4 to 12 weeks of treatment with troglitazone or pioglitazone. Glycemic control was assessed by fasting levels of plasma glucose, hemoglobin A 1c , and serum insulin, and liver function was assessed by asparatate aminotransferase (AST), alanine aminotransferase (ALT), and gamma -glutamyl transpeptidase ( gamma-GTP). Homeostasis model assessment for insulin resistance was used as an index of insulin resistance. During treatment with troglitazone, fasting plasma glucose and hemoglobin A 1c levels and homeostasis model assessment for insulin resistance were significantly decreased. Serum AST, ALT, and gamma-GTP levels were significantly decreased during treatment (AST, -17.4%; ALT, -27.2%; gamma GTP, -47.9%) and returned to pretreatment levels after 4 weeks of withdrawal of the drug. A similar tendency was observed during treatment with pioglitazone (AST, -4.7%; ALT, -16.4%; gamma-GTP, -30.8%). These data suggest that, in contrast to the deterioration of liver function reported in a small subset of patients treated with troglitazone, treatment with thiazolidinediones was associated with a decrease in serum transaminases in most patients. The improvement in liver function parameters known to be associated with fatty liver in the present study, together with an improvement in fatty liver reported for another class of insulin sensitizers, biguanides, suggests that thiazolidinediones may have a beneficial effect on fatty liver. PMID- 15798963 TI - Exercise training is not associated with improved levels of C-reactive protein or adiponectin. AB - The purpose of this study was to determine the effect of exercise training on the levels of C-reactive protein (CRP) and adiponectin, and to assess whether exercise-induced changes in insulin resistance could be explained in part by changes in these inflammation markers. Study participants included 51 middle-aged (45.3+/-8.3 years; mean+/-SD), overweight (33.7+/-4.8 BMI), insulin-resistant, nondiabetic individuals. Subjects had their insulin sensitivity, body fat, CRP, and adiponectin levels measured, and their predicted maximal fitness calculated before and after 16 weeks of moderate, intense, or no exercise training. Modest improvements in fitness, body composition, and insulin sensitivity were observed, but these changes were not associated with decreased CRP or increased adiponectin levels, even when subjects were stratified by their change in fitness or obesity. Regression analysis demonstrated that the change in percentage of body fat was significantly related to changes in insulin sensitivity, whereas changes in VO2 MAX, CRP, and adiponectin were not. Participation in moderate to intense exercise was not associated with improved measures of chronic inflammation markers, as measured by CRP and adiponectin. Moreover, improvements in insulin sensitivity resulting from exercise or modest weight loss did not appear to be related to changes in these markers. PMID- 15798965 TI - The effect of a simultaneous dietary administration of xylitol and ethanol on bone resorption. AB - Our previous studies have shown that dietary xylitol supplementation diminishes bone resorption in rats, as well as protects against ovariectomy-induced increase of bone resorption during experimental osteoporosis. Interestingly, ethanol, when given simultaneously with xylitol, is known to increase blood concentration of xylitol. On the other hand, ethanol, when given alone, has been shown to increase bone resorption. The aim of the present study was to evaluate the effects of a simultaneous dietary administration of 10% xylitol and 10% ethanol on bone resorption. Bone resorption was determined using measurement of urinary excretion of hydrogen 3 (3H) radioactivity in 3H-tetracycline prelabeled rats. Already 4 days after the beginning of dietary supplementations, excretion of 3H was about 15% lower in the xylitol group (diet supplemented with 10% xylitol) and about 25% lower in the xylitol-ethanol group (diet supplemented with 10% xylitol and 10% ethanol) as compared to the controls. The excretion of 3H in these groups remained smaller than that of the controls throughout the entire study period of 40 days. The excretion of 3 H in the xylitol-ethanol group remained also smaller than that of the xylitol group. Bone mineral density and bone mineral content were determined with a peripheral quantitative computed tomography (pQCT) system from the rat tibiae at the end of the experiment. Trabecular bone mineral density and trabecular bone mineral content were significantly greater in the xylitol group and in the xylitol-ethanol group compared to the controls. They were also greater in the xylitol-ethanol group as compared to the xylitol group. Cortical bone mineral density and cortical bone mineral content did not differ significantly between the groups. In conclusion, a simultaneous dietary supplementation with 10% xylitol and 10% ethanol seems to diminish bone resorption and to increase trabecular bone mineral density and trabecular bone mineral content in rats. These effects seem to be stronger than the effects induced by 10% xylitol supplementation alone. PMID- 15798964 TI - Contribution of impaired glucose tolerance in subjects with the metabolic syndrome: Baltimore Longitudinal Study of Aging. AB - BACKGROUND: In addition to fasting plasma glucose (FPG), we examined the contribution of the oral glucose tolerance test (OGTT) in the prevalence of subjects with the metabolic syndrome (MS). METHODS AND RESULTS: Study participants were white adults in the Baltimore Longitudinal Study of Aging who underwent a fasting 2-hour OGTT. In men between the ages of 20 to 39, 40 to 59, 60 to 79, and 80 to 95 years, the prevalence of the MS by Adult Treatment Panel (ATP) III criteria (which excludes OGTT) was 11%, 28%, 32%, and 15%, respectively; whereas in women the prevalence was 5%, 12%, 24%, and 16%, respectively. If the current ATPIII dysglycemia criteria also included a 2-hour postchallenge glucose (2hPG) of 7.8 mmol/L or higher, the prevalence of the MS increased from 25% to 33% in men and from 15% to 21% in women (P<.0001). In study participants with FPG less than 5.6 mmol/L, the prevalence of the MS increased from 16% to 23% in men and from 9% to 13% in women. In men between the ages of 20 to 39, 40 to 59, 60 to 79, and 80 to 95 years and FPG less than 5.6 mmol/L, the prevalence of the MS increased to 15%, 32%, 40%, and 29%, respectively (P<.005 for men between 40 and 95 years of age), with inclusion of an abnormal 2hPG. In women between the ages of 20 to 39, 40 to 59, 60 to 79, and 80 to 95 years and FPG less than 5.6 mmol/L, the prevalence of the MS increased to 7%, 14%, 33%, and 31%, respectively, with inclusion of an abnormal 2hPG (P<.001 for women between 60 and 95 years of age). CONCLUSION: The prevalence of the MS is significantly underestimated when the current ATPIII criteria of FPG 6.1 mmol/L or higher is the only determinant of dysglycemia. PMID- 15798967 TI - Emerging cardiovascular risk factors in subclinical hypothyroidism: lack of change after restoration of euthyroidism. PMID- 15798966 TI - High-sensitivity C-reactive protein is associated with insulin resistance and cardiovascular autonomic dysfunction in type 2 diabetic patients. AB - We tested the hypothesis that elevated levels of plasma high-sensitivity C reactive protein (HSCRP) are associated with insulin resistance/hyperinsulinemia and cardiovascular autonomic dysfunction in type 2 diabetic patients without insulin treatment. The study group consisted of 17 type 2 diabetic patients with high HSCRP (0.3-1.0 mg/dL; age, 59+/-8 years, mean+/-SD; high HSCRP group). The control group consisted of 18 age-matched type 2 diabetic patients with low HSCRP (<0.3 mg/dL; 59+/-7 years; low HSCRP group). Cardiovascular autonomic function was assessed by baroreflex sensitivity, heart rate variability, plasma norepinephrine concentration, and cardiac metaiodobenzylguanidine (MIBG) labeled with iodine 123 scintigraphic findings. Baroreflex sensitivity was lower in the high HSCRP group than in the low HSCRP group (P<.05). Early and delayed 123I-MIBG myocardial uptake values were lower (P<.05 and P<.005, respectively) and the percent washout rate of 123I-MIBG was higher (P<.01) in the high HSCRP group than in the low HSCRP group. Fasting plasma insulin concentration (P<.01) and the homeostasis model assessment index (P<.01) were higher in the high HSCRP group than in the low HSCRP group. Multiple regression analysis revealed that the level of HSCRP was independently predicted by fasting plasma insulin concentration and myocardial uptake of 123I-MIBG at a delayed phase. Our results suggest that high levels of HSCRP are associated with depressed cardiovascular autonomic function and hyperinsulinemia and that fasting plasma insulin concentration and myocardial uptake of 123I-MIBG at a delayed phase are independent predictors of HSCRP level in our Japanese patients with type 2 diabetes mellitus. PMID- 15798968 TI - Carbohydrates as the next frontier in pharmaceutical research. AB - Synthetic carbohydrates and glycoconjugates are used to study their roles in biological important processes such as inflammation, cell-cell recognition, immunological response, metastasis, and fertilization. The development of an automated oligosaccharide synthesizer greatly accelerates the assembly of complex, naturally occurring carbohydrates as well as chemically modified oligosaccharide structures and promises to have major impact on the field of glycobiology. Tools such as microarrays, surface plasmon resonance spectroscopy, and fluorescent carbohydrate conjugates to map interactions of carbohydrates in biological systems are presented. Case studies of the successful application of carbohydrates as active agents are discussed, for example, fully synthetic oligosaccharide vaccines to combat tropical diseases (e.g., malaria), bacterial infections (e.g., tuberculosis), viral infections such as HIV, and cancer. Aminoglycosides serve as examples of drugs acting through carbohydrate-nucleic acid interactions, while heparin works by carbohydrate-protein interactions. A general, modular strategy for the complete stereoselective synthesis of defined heparin oligosaccharides is presented. A carbohydrate-functionalized fluorescent polymer has been shown to detect miniscule amounts of bacteria faster than commonly used methods. PMID- 15798969 TI - Donor-substituted cyanoethynylethenes: pi-conjugation and band-gap tuning in strong charge-transfer chromophores. AB - An extensive series of silyl-protected cyanoethynylethenes (CEEs) and N,N dimethylanilino donor-substituted CEEs have been synthesized. More extended chromophores were constructed by selective silyl deprotection and subsequent oxidative acetylenic coupling. The strong electron-accepting nature of the CEEs was revealed by a combination of 13C NMR spectroscopic and electrochemistry measurements. Donor-substituted CEEs display strong intramolecular charge transfer (CT) character, resulting in intense, bathochromically shifted CT bands in the UV/Vis spectrum. Their structural diversity establishes them as suitable models for the study of pi-conjugation and band gap tuning in strong charge transfer chromophores. The extent of pi-conjugation in the donor-substituted CEEs was investigated by a combination of ground-state techniques, such as X-ray crystallography, electrochemistry, B3 LYP calculations, and NMR spectroscopy. The comparison of these ground-state results with the features observed in the UV/Vis spectra reveals that-contrary to expectations-more extensive pi-conjugation can lead to larger band gaps in molecules with strong donor and acceptor moieties. PMID- 15798970 TI - Giant meso-meso-linked porphyrin arrays of micrometer molecular length and their fabrication. AB - On the basis of the Ag(I)-promoted coupling reaction of zinc(II)-5,15-bis(3,5 dioctyloxyphenyl)porphyrin Z1, chain elongation has been attempted by using a stepwise doubling approach, which provides Z2, Z4, Z8, Z16, Z32, Z64, Z128, Z256, Z384, and Z512. The porphyrin arrays up to Z128 are sufficiently soluble in CHCl3 and THF despite their very long molecular lengths and rodlike structures, while the arrays over Z128 show a significant drop in solubility and stability. The discrete porphyrin arrays thus isolated were characterized by means of (1)H NMR spectroscopy, matrix-assisted laser desorption ionization time-of-flight (MALDI TOF) mass spectrometry, UV/Vis spectroscopy, gel-permeation chromatography (GPC), cyclic voltammetry (CV), single-crystal X-ray crystallography, scanning tunneling microscopy (STM), and atomic force microscopy (AFM). Contrary to expected linear conformations of the arrays Z n (where n is the number of porphyrins), the single molecular images of Z128, Z256, and Z512 revealed largely bent structures; this finding indicates the substantial conformational flexibility of Z n. We also exploited an effective synthetic route by means of which Z n can be fabricated with a thiol-protected aryl group to provide Z n S(2) through Z n Br(2), by bromination with N-bromosuccinimide and subsequent Pd-catalyzed Suzuki-Miyaura arylation. Finally, the reaction of Z256 provided Z512, Z768, and Z1024. Collectively, this work provides an important milestone in the preparation of sub microscale discrete organic molecules and the fabrication of molecular-based materials, hence significantly contributing to device applications. PMID- 15798971 TI - Synthesis of mono- and disubstituted porphyrins: A- and 5,10-A2-type systems. AB - General syntheses have been developed for meso-substituted porphyrins with one or two substituents in the 5,10-positions and no beta substituents. 5-Substituted porphyrins with only one meso substituent are easily prepared by an acid catalyzed condensation of dipyrromethane, pyrrole-2-carbaldehyde, and an appropriate aldehyde using a "[2+1+1]" approach. Similarly, 5,10-disubstituted porphyrins are accessible by simple condensation of unsubstituted tripyrrane with pyrrole and various aldehydes using a "[3+1]" approach. The yields for these reactions are low to moderate and additional formation of either di- or monosubstituted porphyrins due to scrambling of the intermediates is observed. However, the reactions can be performed quite easily and the desired target compounds are easily removed due to large differences in solubility. A complementary and more selective synthesis involves the use of organolithium reagents for S(N)Ar reactions. Reaction of in situ generated porphyrin (porphine) with 1.1-8 equivalents of RLi gave the monosubstituted porphyrins, while reaction with 3-6 equivalents of RLi gave the 5,10-disubstituted porphyrins in yields ranging from 43 to 90 %. These hitherto almost inaccessible compounds complete the series of different homologues of A-, 5,15-A(2)-, 5,10-A(2)-, A(3)-, and A(4) type porphyrins and allow an investigation of the gradual influence of type, number, and regiochemical arrangement of substituents on the properties of meso substituted porphyrins. They also present important starting materials for the synthesis of ABCD porphyrins and are potential synthons for supramolecular materials requiring specific substituent orientations. PMID- 15798972 TI - Intramolecular electron transfer between tyrosine and tryptophan photosensitized by a chiral pi,pi* aromatic ketone. AB - The photochemical reaction of Trp and Tyr and related peptides with Suprofen (SUP) as sensitizer in H2O/CH3CN (28:1 v/v) solutions has been studied by time resolved spectroscopy. The results show that SUP induces oxidation of both Trp and Tyr, as well as intramolecular-ET reactions in the related peptides. The influence of photosensitizer configuration on the involved processes has been studied by using the enantiomerically pure compounds. A significant chiral recognition is observed in which the concentration of the radicals formed after triplet quenching depends on the configuration of the chiral center; the quenching process is higher when using the (R)-SUP enantiomer. PMID- 15798973 TI - Synthesis and supramolecular properties of molecular clips with anthracene sidewalls. AB - Novel molecular clips with anthracene sidewalls (1 a-c) were synthesized; they form stable host-guest complexes with a variety of electron-deficient aromatic and quinoid molecules. According to single-crystal structure analyses of clip 1 c and 1,2,4,5-tetracyanobenzene (TCNB) complex 14@1 b, the clips' anthracene sidewalls have to be compressed substantially during the complex formation to provide attractive pi-pi interactions between the aromatic guest molecule and the two anthracene sidewalls in the complex. The compression and expansion of aromatic sidewalls are calculated by molecular mechanics to be low-energy processes, so the energy required for compression of the anthracene sidewalls during complex formation is apparently overcompensated by the gain in energy resulting from the attractive pi-pi interactions. The finding that complexes of the clips 1 a-c are more stable than those of the corresponding clips 2 a-c can be explained in terms of the larger van der Waals contact surfaces of the anthracene sidewalls in 1 a-c (relative to the naphthalene sidewalls in 2 a-c). Color changes resulting from charge-transfer (CT) bands are observed in complex formation by 1 a-c: from colorless to red or purple with TCNB (14), and from yellow to green with 2,4,7-trinitro-9-fluorenone TNF (17). Independently, the host 1 b and guest 14 fluoresce from their respective excited singlet states, whilst in the complex 14@1 b the charge-transfer state quenches the higher-energy singlet states of the two components, and as a result luminescence is only observed from this new CT state. To the best of our knowledge, complex 14@1 b is the first example of CT luminescence from a host-guest complex. The binding constant determined for the formation of the TCNB complex 14@1 b from a UV/Vis titration experiment (Ka = 12 400 m(-1)) agrees well with the value (K(a) = 12 800 m(-1)) obtained by 1H NMR titration. PMID- 15798974 TI - Convenient synthesis of multifunctional EDTA-based chiral metal chelates substituted with an S-mesylcysteine. AB - We describe the synthetic route to ethylenediaminetetraacetic acid (EDTA) derivatives that can be attached to surface-exposed thiol functional groups of cysteine residues in proteins, via a methylthiosulfonate moiety that is connected in a stereochemically unique way to the C-1 carbon atom of EDTA. Such compounds can be used to align proteins in solution without the need to add liquid crystalline media, and are, therefore, of great interest for the NMR spectroscopic analysis of biomolecules. The binding constant for the paramagnetic tag to lanthanide ions was determined by measuring luminescence. For the Tb(+3) ligand complex, a K(b) value of 6.5 x 10(17) M(-1) was obtained. This value is in excellent agreement with literature values for the related EDTA compound. In addition, it could be shown that there is no significant reduction in the luminescence intensity upon addition of a 10(4) excess of Ca2+ ions, indicating that this paramagnetic tag is compatible with buffers containing high concentrations of divalent alkaline earth ions. PMID- 15798975 TI - The kinetic acidity of oligofluorobenzenes correlated with their gas phase deprotonation energies. AB - The relative reactivities of fluorobenzene, all di-, tri-, and tetrafluorobenzenes and pentafluorobenzene toward sec-butyllithium have been assessed in tetrahydrofuran at -100 degrees C. At this temperature no subsequent transmetalation reactions take place but those compromise the outcome of the competition experiments if the latter are conducted at -75 degrees C. The rates determined at -100 degrees C reflect the basicity differences between the naked (oligo)fluorophenyl anions to the extent of 10 %. PMID- 15798976 TI - High charge-carrier mobility in pi-deficient discotic mesogens: design and structure-property relationship. AB - Hexaazatrinaphthylene (HATNA) derivatives with six alkylsulfanyl chains of different length (hexyl, octyl, decyl and dodecyl) have been designed to obtain new potential electron-carrier materials. The electron-deficient nature of these compounds has been demonstrated by cyclic voltammetry. Their thermotropic behaviour has been studied by means of differential scanning calorimetry and polarised optical microscopy. The supramolecular organisation of these discotic molecules has been explored by temperature-dependent X-ray diffraction on powders and oriented samples. In addition to various liquid crystalline columnar phases (Col(hd), Col(rd)), an anisotropic plastic crystal phase is demonstrated to exist. The charge-carrier mobilities have been measured with the pulse-radiolysis time-resolved microwave-conductivity technique. They are found to be higher in the crystalline than in the liquid crystalline phases, with maximum values of approximately 0.9 and 0.3 cm(2) V(-1) s(-1), respectively, for the decylsulfanyl derivative. Mobilities strongly depend on the nature of the side chains. PMID- 15798977 TI - On the reaction of Ph2PNHPPh2 with RNCS (R=Et, Ph, p-NO2C6H4): preparation of the zwitterionic ligand EtNHC(S)Ph2P==NPPh2C(S)NEt (HSNS) and the zwitterionic metalate [(SNS)Rh(CO)]. AB - The reaction of Ph(2)PNHPPh(2) (PNP) with RNCS (Et, Ph, p-NO(2)(C(6)H(4))) gives addition products resulting from the attack of the P atoms of PNP on the electrophilic carbon atom of the isothiocyanate. When PNP is reacted with EtNCS in a 1:2 molar ratio, the zwitterionic molecule EtNHC(S)PPh(2)==NP(+)Ph(2)C(S)N( )Et (HSNS) is obtained in high yield. HSNS can be protonated (H(2)SNS(+)) or deprotonated (SNS(-)), behaving in the latter form as an S,N,S-donor pincer ligand. The reaction of HSNS with [(acac)Rh(CO)(2)] (acac=acetylacetonate) affords the zwitterionic metalate [(SNS)Rh(CO)]. Other products can be obtained depending on the R group, the PNP/RNCS ratio (1:1 or 1:2), and the reaction temperature. The proposed product of the primary attack of PNP on RNCS, Ph(2)PN==PPh(2)C(S)NHR (A), cannot be isolated. Reaction of A with another RNCS molecule leads to 1:2 addition compounds of the general formula RNHC(S)PPh(2)==NP(+)Ph(2)C(S)N(-)R (1), which can rearrange into the non zwitterionic product RNHC(S)PPh(2)==NP(S)Ph(2) (2) by eliminating a molecule of RNC. Two molecules of A can react together, yielding 1:1 PNP/RNCS zwitterionic products of the formula RNHCH[PPh(2)==NP(S)Ph(2)]PPh(2)==NP(+)Ph(2)C(S)N(-)R (3). Compound 3 can then rearrange into RNHCH[PPh(2)==NP(S)Ph(2)](2) (4) by losing a RNC molecule. When R=Et (a), compounds 1 a, 2 a (HSNS), and 4 a have been isolated and characterized. When R=Ph (b), compounds 2 b and 4 b can be prepared in high yield. When R=p-NO(2)C(6)H(4) (c), only compound 3 c is observed and isolated in high yield. The crystal structures of HSNS, [(SNS)Rh(CO)], and of the most representative products have been determined by X-ray diffraction methods. PMID- 15798979 TI - Polymer-induced alignment of DL-alanine nanocrystals to crystalline mesostructures. AB - Crystallization of DL-alanine by cooling of a supersaturated solution in the presence of a chiral double-hydrophilic block copolymer poly(ethylene glycol) block-poly(ethylene imine)-S-isobutyric acid (PEG4700-PEI1200-S-iBAc) yields crystal superstructures with an astonishing morphology. Although analysis by light microscopy reveals these crystals to be apparently well facetted, electron microscopy shows that they consist of three-dimensionally, well-aligned nanocrystals that are scaffolded to a so-called mesocrystal. This mesocrystal is formed by polymer-mediated structuration, and provides evidence for the importance of mesoscopic events in a typical crystallization process. PMID- 15798978 TI - Total synthesis of (-)-laulimalide: Pd-catalyzed stereospecific ring construction of the substituted 3,6-dihydro[2H]pyran units. PMID- 15798980 TI - Asymmetric amplification using chiral cocrystals formed from achiral organic molecules by asymmetric autocatalysis. PMID- 15798981 TI - Redox-induced transformation from an extended to a pi-stacked conformer in acyclic bis(catecholacetal)s of acetylacetone. PMID- 15798982 TI - Monodisperse magnetic single-crystal ferrite microspheres. PMID- 15798983 TI - Arthur Eichengrun--tribute to a forgotten chemist, entrepreneur, and German Jew. PMID- 15798984 TI - Discotic liquid crystals with an inverted structure. PMID- 15798985 TI - Quantitative dynamic interconversion between Ag(I)-mediated capsule and cage complexes accompanying guest encapsulation/release. PMID- 15798986 TI - Chasing a phantom by total synthesis: the butylcycloheptylprodigiosin case. PMID- 15798987 TI - Wound closure in the invasive green alga Caulerpa taxifolia by enzymatic activation of a protein cross-linker. PMID- 15798988 TI - Scaling the alignment of small organic molecules in substituted polyglutamates by variable-angle sample spinning. PMID- 15798989 TI - One-nanometer-scale size-controlled synthesis of monodisperse magnetic iron oxide nanoparticles. PMID- 15798990 TI - A structure-based strategy for discovery of small ligands binding to functionally unknown proteins: combination of in silico screening and surface plasmon resonance measurements. AB - In the postgenomic era, many researchers and organizations have been engaged in structural and functional analyses of proteins. As a part of these efforts, searching for small organic compounds that bind specifically to target proteins is quite important. In this study, we have developed a rational strategy for ligand discovery based on the three-dimensional structures of target proteins, which were elucidated by X-ray crystallography and nuclear magnetic resonance spectroscopy. The strategy has three features: (i) rapid selection of candidate compounds by in silico screening, (ii) automated preparation of sample solutions with robotics, and (iii) reliable evaluation of the candidates with surface plasmon resonance. Applying the strategy to a protein, At2g24940 from Arabidopsis thaliana, we discovered four small ligands out of a commercially available library of about 150 000 compounds. Although these compounds had only weak affinities to the target protein, with dissociation constants ranging from 68 to 120 microM, they apparently possess common structural features. They would be leads for the development of specific inhibitors/drugs for At2g24940, and provide important clues toward elucidation of the protein function. PMID- 15798991 TI - Development of a lectin microarray for the rapid analysis of protein glycopatterns. PMID- 15798992 TI - Changes in serum alpha2u-globulin levels in castrated male rats treated with testosterone propionate in a Hershberger assay. AB - The Hershberger assay has been proposed as a candidate screening test method for the detection of androgenic and anti-androgenic chemicals and is being validated presently under the test guideline programme of the Organization for Economic Cooperation and Development (OECD). Rat alpha2u-globulin is male rat-specific protein appearing in their serum and urine, and the protein is known to be induced by androgens. We investigated the usefulness of measuring serum alpha2u globulin levels as a parameter for the androgenic activity of chemicals tested in the Hershberger assay. The serum alpha2u-globulin level was measured after the administration of testosterone propionate at dosages of 0, 20, 100 or 500 microg kg(-1) day(-1) for ten consecutive days in the castrated male rats. The ventral prostate, balbocavernosus/levator ani muscles, glans penis and Cowper's gland were collected and weighed. Although all the androgen-responsive organ weights were increased significantly at dosages of 100 and 500 microg kg(-1) day(-1), the serum alpha2u-globulin level was increased significantly only at a dosage of 500 microg kg(-1) day(-1). These results show that the serum alpha2u-globulin level may be a useful biomarker for detecting androgenic activity caused by test chemicals, but it is less sensitive than the organ weights of androgen-responsive tissues in the Hershberger assay. PMID- 15798993 TI - Discovering high mobility group A molecular partners in tumour cells. AB - DNA-based activities rely on an extremely coordinated sequence of events performed by several chromatin-associated proteins which act in concert. High Mobility Group A (HMGA) proteins are non-histone architectural nuclear factors that participate in the regulation of specific genes but they are also believed to have a more general role in chromatin dynamics. The peculiarity of these proteins is their flexibility, both in terms of DNA-binding and in protein protein interactions. Since these proteins act as core elements in the assembly of multiprotein complexes called enhanceosomes, and have already displayed the ability to interact with several different proteins, we started a proteomic approach for the systematic identification of their molecular partners. By a combination of affinity chromatography, two-dimensional gel electrophoresis and mass spectrometry we have identified about twenty putative HMGA interactors which could be roughly assigned to three different classes: mRNA processing proteins, chromatin remodelling related factors and structural proteins. Direct HMGA interaction with some of these proteins was confirmed by glutathione-S transferase pull-down assays and the HMGA domain involved was mapped. Blot overlay experiments reveal that members of the HMGA family share most of their molecular partners but, interestingly, it seems that there are some cell-type specific partners. Taken together, these experimental data indicate that HMGA proteins are highly connected nodes in the chromatin protein network. Since these proteins are strongly implicated with cancer development, the identification of molecules able to perturb the HMGA molecular network could be a possible tool to interfere with their oncogenic activity. PMID- 15798994 TI - Guggulu (Commiphora mukul) potentially ameliorates hypothyroidism in female mice. AB - The efficacy of guggulu, the gum resin of Commiphora mukul in regulating hypothyroidism was evaluated in female mice. In addition to estimating serum levels of thyroxine and triiodothyronine, hepatic 5' monodeiodinase, hepatic glucose-6-phospatase and lipid-peroxidation (LPO), the activities of the anti oxidative enzymes, superoxide dismutase (SOD) and catalase (CAT), were investigated. While 6-n-propyl-2-thiouracil (PTU, 10.00 mg/kg/d for 30 days) induced hypothyroidism in mice, as evidenced by a decrease in thyroid hormone concentration and in hepatic 5'D-I activity, simultaneous administration of guggulu (200 mg/kg/d for 30 days) reversed this effect, indicating its potential to stimulate thyroid function. Although in PTU treated animals a marginal increase in hepatic LPO was observed, when simultaneously treated with guggulu, it was decreased. A parallel increase in the activity of endogenous antioxidants, SOD and CAT, in the latter group indicated the safe and antiperoxidative nature of the drug. These findings suggest the possible use of guggulu in the amelioration of hypothyroidism. PMID- 15798995 TI - Bioactive triterpenoids from Vochysia pacifica interact with cyclic nucleotide phosphodiesterase isozyme PDE4. AB - Several species of the genus Vochysia (Vochysiaceae) are used by traditional communities in South America to relieve ailments linked to inflammation, such as skin sores, asthma and pulmonary congestion. As the cAMP phosphodiesterase 4 isozyme (PDE4) is currently considered as an intracellular target for new antiinflammatory drugs, several constituents of Vochysia pacifica Cuatrec., an endemic tree from the Western coast of Colombia, were tested for their ability to inhibit PDE4. Purification of the methanol extract of the stem bark of this species led to the isolation of seven known triterpene derivatives: betulinic acid, sericic acid, 24-hydroxytormentic acid, trachelosperogenin B, 24 hydroxytormentic acid glucosyl ester, quadranoside I and niga-ichigoside F1. One triterpene glycoside, quadranoside I, and two triterpenes, betulinic and sericic acids, exhibited mild inhibitory activity on the isolated PDE4 isozyme. PMID- 15798997 TI - Clinical toxicology study of an herbal medicinal extract of Paullinia cupana, Trichilia catigua, Ptychopetalum olacoides and Zingiber officinale (Catuama) in healthy volunteers. AB - In Brazil, a herbal medicinal extract named Catuama containing a mixture of Paullinia cupana (guarana; Sapindaceae), Trichilia catigua (catuaba; Meliaceae), Ptychopetalum olacoides (muirapuama; Olacaceae) and Zingiber officinale (ginger; Zingiberaceae) is used as a body stimulant, energetic, tonic and aphrodisiac. The present study investigated the chronic administration of 25 mL Catuama twice a day during 28 days for any toxic effect on healthy human volunteers of both sexes. No severe adverse reactions or haematological and biochemical changes were reported. PMID- 15798996 TI - In vitro antimycotic activity of some plant extracts towards yeast and yeast-like strains. AB - As part of screening aimed at the selection of novel antimycotic compounds of vegetable origin, leaf extracts of Camellia sinensis L., Cupressus sempervirens L. and Pistacia lentiscus L. and the seed extract of Glycine soja Sieb. et Zucc. were tested against yeast and yeast-like species implicated in human mycoses. Of the extracts only those of C. sinensis (obtained from a commercial preparation of green tea) exhibited broad activity towards Candida glabrata, Clavispora lusitatiae, Cryptococcus laurentii, Filobasidiella neoformans, Issatchenkia orientalis, Saccharomyces cerevisiae and Prototheca wickerhamii strains. MICs ranging from 300 to 4800 microg extract/mL (corresponding to 130-2010 microg/mL total polyphenols) were observed. Concentrations of the C. sinensis extract over 25 000 microg/mL caused a rapid decrease of viable cells of Fil. neoformans and its activity was dose-dependent. Tests carried out using the pure polyphenols present in C. sinensis extract composition, showed that only epicatechin-3-O gallate (ECG) and epigallocatechin-3-O-gallate (EGCG) possess antimycotic activity. PMID- 15798998 TI - Inhibitory effects of stilbenes in Sophora moorcroftiana BENTH ex BAKER on copper ion-induced protein oxidative modification of mouse brain homogenate in vitro. AB - We present the results of an in vitro investigation of the inhibitory effects of sophorastilbene A and (+)-alpha-viniferin isolated from Sophora moorcroftiana BENTH ex BAKER on copper ion-induced protein oxidative modification. They inhibited copper-induced protein oxidative modification. The order of these stilbenes and mannitol as a hydroxyl radical scavenger was sophorastilbene A > (+)-alpha-viniferin > mannitol. PMID- 15798999 TI - Potential biomedical properties of Pinus massoniana bark extract. AB - This study examined the potential biomedical properties of a bark extract from Pinus massoniana Lamb (PMBE). Its antioxidant activity, superoxide anion radical scavenging and free radical scavenging activities were examined. The antioxidant activity in a linoleic acid emulsion increased with increasing amounts of PMBE, with 100, 300 and 500 microg PMBE inhibiting 64.7%, 68.6% and 76.6% of peroxidation, respectively. Similarly, the radical scavenging activity by PMBE increased in a dose-dependent manner, suggesting that there may be a correlation between the antioxidant and scavenging activities. In addition, the effect of PMBE on human hepatocellular carcinoma BEL-7402 cells and normal liver L-02 cells was investigated in vitro. PMBE appeared to inhibit selectively the growth of BEL 7402 and slightly promoted the growth of L-02. Taken together, the results indicate that PMBE is a natural antioxidant that could potentially be used as a food supplement or as a candidate precursor substance for new anticancer therapeutics. PMID- 15799001 TI - Evaluation of chemopreventive action and antimutagenic effect of the standardized Panax ginseng extract, EFLA400, in Swiss albino mice. AB - In the present investigation the chemopreventive action and antimutagenic effects of a standardized Panax Ginseng extract (EFLA400, processed Panax ginseng extract containing a high titre of ginsenoside Rg3 (>3.0% w/w) known as Phoenix ginseng) in Swiss albino mice have been evaluated. The oral administration of EFLA400 at 1, 3 and 10 mg/kg body weight at pre, peri and post-initiational phases, showed significant reductions in the number, size and weight of the papillomas. A significant reduction in tumour incidence (71.41 +/- 6.73%, 72.19 +/- 4.54% and 70.46 +/- 0.38% at 1, 3 and 10 mg/kg body weight, respectively) was observed in animals in the EFLA400 treated group compared with 100% tumour incidence in the control group. The cumulative number of papillomas during an observation period of 16 weeks was significantly reduced in the EFLA400 treated group (24 +/- 0.94, 16 +/- 1.41 and 11 +/- 1.41 at 1, 3 and 10 mg/kg body weight, respectively). However, the average latent period was significantly increased from 10.81 +/- 0.1 weeks in the control group to 12.39 +/- 0.28 weeks in the treated group (10 mg/kg body weight). The average tumour weight was recorded as 128.55 +/- 8.48, 116.00 +/- 8.48 and 57.5 +/- 3.29 mg in 1, 3 and 10 mg/kg body weight EFLA400 treated groups respectively. Chromosomal aberrations and micronuclei induction was also evaluated in bone marrow cells. These genotoxicity end-points were compared with papilloma occurrence at the same dose levels of carcinogen and ginseng. In the EFLA400 treated groups significantly reduced frequencies of chromosomal aberrations and micronuclei induced by DMBA and croton oil were observed. However, the maximum decrease in the frequencies of chromosomal aberrations and micronuclei were recorded in the 10 mg/kg body weight EFLA400 treated group than that of the 1 and 3 mg/kg body weight EFLA400 treated animals. The results from the present study suggest the dose dependent effectiveness of EFLA400 in chemoprevention and antimutagenicity in Swiss albino mice. PMID- 15799000 TI - Evaluation of antioxidant potential of the cultured mycobiont of a lichen Usnea ghattensis. AB - The biologically active compounds, antioxidant activities and free radical scavenging effects of dry matter of cultural medium (DMCM), filtrate (DMF) and different solvent extracts of mycobiont from lichen species Usnea ghattensis in submerged culture (UGSC) were investigated. DMF showed the strongest inhibition of lipid peroxidation as a function of its concentration, and was comparable to the antioxidant activity of BHA at the same concentration of 0.2 mg/mL. The hexane extract of mycobiont had the weakest antioxidant ability, whereas other mycobiont extracts exhibited a modest to strong inhibition of lipid peroxidation and in free radical scavenging activity. The antioxidant activities of mycobiont extracts were correlated with the presence of total polyphenol content in the extract. It was found that antioxidant components in methanol, acetone, ethanol and hexane extracts must be derived from the secondary metabolites produced by the lichen mycobiont. The results presented herein indicate that extracts of cultured mycobiont of lichen Usnea ghattensis could possibly act as chemopreventing agent with respect to free radical-related diseases. PMID- 15799002 TI - Reversal of LPS-induced central and peripheral hyperalgesia by green tea extract. AB - Tea has recently attracted a great deal of attention for its beneficial health effects. Green tea polyphenols inhibit the production of arachidonic acid metabolites and leukotrienes resulting in decreased inflammatory responses. In the present study, the effect of green tea extract (GTE) on lipopolysaccharide (LPS)-induced thermal and behavioural hyperalgesia in mice and the possible involvement of the cyclooxygenase pathway in this paradigm was evaluated. GTE (25 mg/kg, i.p.), nimesulide (2 mg/kg, i.p.) and rofecoxib (2 mg/kg, i.p.) significantly attenuated LPS-induced thermal and behavioural hyperalgesia but per se did not modify any of the behavioural effects. Concurrent administration of a subeffective dose of GTE (10 mg/kg, i.p.) and rofecoxib (2 mg/kg, i.p.) or nimesulide (2 mg/kg, i.p.) significantly potentiated the antinociceptive effect of GTE in both LPS-induced thermal and behavioural hyperalgesia with nimesulide showing a more pronounced enhancing effect. Thus it can be concluded that GTE attenuates LPS-induced central and peripheral hyperalgesia by selective inhibition of cyclooxygenase-2 enzyme. PMID- 15799003 TI - Fucoidan inhibits the development of proteinuria in active Heymann nephritis. AB - Fucoidan, the sulphated polysaccharide extracted from brown seaweed, has various biological activities. The effect of fucoidan on the formation of proteinuria and renal functions in active Heymann nephritis was investigated in this study. Active Heymann nephritis was induced by administering brush border protein of rat proximal uriniferous tubules (FX1A). Fucoidan was administered by oral intubation to Heymann nephritis rats at three doses (50, 100 and 200 mg/kg) once daily for 4 weeks. The elevated urinary protein excretion and plasma creatinine due to the induction of Heymann nephritis were significantly reduced by fucoidan at doses of 100 and 200 mg/kg. The results indicated that fucoidan has a renoprotective effect on active Heymann nephritis and is a promising therapeutic agent for nephritis. PMID- 15799004 TI - Protective value of Aloe vera against some toxic effects of arsenic in rats. AB - Concomitant oral supplementation of Aloe vera, (1, 2 or 5% w[sol ]v in drinking water) during arsenic exposure (0.2 mg[sol ]kg, intraperitoneally, once daily for 3 weeks) was investigated in rats for its protective value. Animals exposed to arsenic (III) showed a significant inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity, a marginal decrease in glutathione (GSH) and an increase in zinc protoporphyrin (ZPP) level in blood. White blood corpuscles (WBC) level decreased while most of the other clinical blood parameters like red blood cells count, haemoglobin, MCV, MCH, MCHC ratio and platelet number, etc. remained unaltered on arsenic exposure. Hepatic reduced GSH, oxidized glutathione (GSSG) level remained unaltered, thiobarbituric acid reactive substance (TBARS) level increased significantly while the activity of alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and catalase decreased on arsenic exposure. Renal GSH contents decreased while superoxide dismutase (SOD) activity decreased significantly on arsenic exposure. Concomitant administration of Aloe vera had remarkable protective action on inhibited blood ALAD activity and restored blood GSH level while most of the other blood biochemical parameters remained unchanged on Aloe vera supplementation. Interestingly, most of hepatic biochemical variables indicative of oxidative stress showed protection; no effect of Aloe vera on blood and liver arsenic concentration was noted. Also, no effect of Aloe vera on most of the altered renal biochemical parameters were noticed. The results thus lead us to conclude that simultaneous supplementation of Aloe vera protects against arsenic induced oxidative stress but does not influence the arsenic concentration in these organs. PMID- 15799005 TI - Antiallergic and antihistaminic effect of two extracts of Capparis spinosa L. flowering buds. AB - The antiallergic properties of two lyophilized extracts obtained from Capparis spinosa L. flowering buds (capers) by methanol extraction, carried out at room temperature (CAP-C) or with heating at 60 degrees C (CAP-H), were investigated. The protective effects of CAP-H and CAP-C, orally administered (14.28 mg[sol ]kg), were evaluated against Oleaceae antigen challenge-induced and histamine induced bronchospasm in anaesthetized guinea-pigs. Furthermore, the histamine skin prick test was performed on humans, applying a gel formulation containing 2% CAP-C (the only extract able to protect against histamine-induced bronchospasm) on the skin for 1 h before histamine application and monitoring the erythema by reflectance spectrophotometry. The CAP-H showed a good protective effect against the bronchospasm induced by antigen challenge in sensitized guinea-pigs; conversely, a significant decrease in the responsiveness to histamine was seen only in CAP-C pretreated animals. Finally, the CAP-C gel formulation possessed a marked inhibitory effect (46.07%) against histamine-induced skin erythema. These two caper extracts displayed marked antiallergic effectiveness; however, the protective effect of CAP-H was very likely due to an indirect mechanism (for example, inhibition of mediator release from mast cells or production of arachidonic acid metabolites); conversely, CAP-C is endowed with direct antihistaminic properties. The different mechanisms of action of CAP-H and CAP-C may be related to a difference in the extraction procedure and, thus, in their qualitative[sol ]quantitative chemical profile. PMID- 15799006 TI - Bioequivalence of two recombinant granulocyte colony-stimulating factor formulations in healthy male volunteers. AB - To evaluate the equivalence of the pharmacokinetic, pharmacodynamic and safety properties of two recombinant G-CSF formulations in healthy male volunteers, a standard 2-way randomized crossover double-blind study, with a 3 week washout period, was conducted. A single 300 microg G-CSF dose was administered subcutaneously. Hebervital (Heber Biotec, Havana, formulation A) and Neupogen (Hoffmann-La Roche S.A, formulation B) were compared. Twenty-four healthy male volunteers were included. The serum G-CSF level was measured by enzyme immunoassay (EIA) during the first 36 h after administration. Absolute neutrophils (ANC), white blood cells (WBC) and CD34+ cells counts were the pharmacodynamic variables measured up to 120 h. Other clinical and laboratory determinations were used as safety criteria. The pharmacokinetic parameters for formulation A and B were very close to each other (i.e. AUC, 235.9 vs 270.0 ng.h/ml; C(max), 29.2 vs 33.4 ng/ml; T(max), 4.2 vs 4.7 h; half-life, 3.2 vs 2.8 h; CL, 260.9 vs 277.2 ml/h; V(d), 1.2 vs 1.1 l; and MRT, 7.58 vs 7.38 h). The confidence intervals for the means ratio of all these parameters were within or very close to the 0.8-1.25 acceptance range. The pharmacodynamics showed high similarity since ANC and WBC had the same profiles for both products and no differences were detected for the estimated parameters. The CD34+ cells count increments were evident for both formulations in a similar way as well. The treatments were well tolerated. Registered adverse events were similar; back/spine pain was the most frequent. According to the overall results these formulations could be considered as clinically comparable. PMID- 15799007 TI - Radioprotection by plant products: present status and future prospects. AB - The development of radioprotective agents has been the subject of intense research in view of their potential for use within a radiation environment, such as space exploration, radiotherapy and even nuclear war. However, no ideal, safe synthetic radioprotectors are available to date, so the search for alternative sources, including plants, has been on going for several decades. In Ayurveda, the traditional Indian system of medicine, several plants have been used to treat free radical-mediated ailments and, therefore, it is logical to expect that such plants may also render some protection against radiation damage. A systematic screening approach can provide leads to identifying potential new candidate drugs from plant sources, for mitigation of radiation injury. This article reviews some of the most promising plants, and their bioactive principles, that are widely used in traditional systems of medicine, and which have rendered significant radioprotection in both in vitro and in vivo model systems. Plants and their constituents with pharmacological activities that may be relevant to amelioration of radiation-mediated damage, including antiemetic, antiinflammatory, antioxidant, cell proliferative, wound healing and haemopoietic stimulatories are also discussed. PMID- 15799008 TI - Comments on the paper 'The effect of a herbal medicine (Mao-to) for the prevention of interferon-induced psychiatric complications in chronic hepatitis C: a pilot study'. PMID- 15799010 TI - Reboxetine for the treatment of patients with Cocaine Dependence Disorder. AB - BACKGROUND: Although several approaches have been attempted for cocaine dependence, the pharmacological treatment of this serious disorder remains unclear. To date, desipramine, a tricyclic antidepressant of great noradrenergic activity, has shown the best results. Reboxetine, a selective noradrenaline reuptake inhibitor, might be an effective therapeutic option for this severe drug addiction. The aim was preliminarily to assess reboxetine in a group of cocaine dependent patients, selected from The Madrid City Council Drug Addiction Program primary care centres. METHOD: Twenty six patients with a diagnosis of cocaine dependence disorder (DSM-IV 304.20) were selected to receive open treatment with reboxetine, 8 mg/day, for 12 weeks. Follow up assessments comprised cocaine consumption, treatment retention rate and change in standard structured psychometric instrument scores: cocaine selective severity assessment, Hamilton anxiety scale, Hamilton depression scale and clinical global impression, throughout the treatment period. RESULTS: Data were obtained from 20 patients; 10 of them remained abstinent, whereas the other 10 consumed cocaine at some time during the study. The treatment retention rate at week 12 was 61.5%. The psychometric instrument mean scores showed marked decreases throughout the treatment period. CONCLUSION: Reboxetine might be an effective and safe therapeutic option for cocaine dependence disorder. The aversive effects, as well as the high blockage reported by some patients consuming cocaine during the trial, might be related to treatment. If confirmed in large clinical trials, the trends suggested by this study would confirm the role of noradrenergic function in the treatment of cocaine dependence. PMID- 15799012 TI - Antifungal activity and studies on mode of action of novel xanthoxyline-derived chalcones. AB - Chalcones and chalcone-like compounds, most of them new ones, prepared by base catalyzed condensation of appropriate aldehydes and xanthoxyline, were tested for antifungal properties against a panel of yeasts, hialohyphomycetes as well as dermatophytes with the agar dilution assay. Results indicate that neither the sole presence of a "xanthoxyline-like" substitution pattern nor a 2'-OH substituent on ring A are sufficient for these compounds to have antifungal properties. The chalcone 3-(2-chlorophenyl)-1-(2'-hydroxy-4',6' dimethoxyphenyl)prop-2-en-1-one, with a Cl atom in the ortho position of benzene ring B showed the best antifungal activity against standardized strains of Trichophyton rubrum (MIC = 12.5 microg/mL) and inhibited all of the ten clinical isolates of T. rubrum tested (MIC at which 50 % [MIC(50)] and 90 % [MIC(90)] of the isolates were inhibited = 12.5 and 25 microg/mL). Regarding its mode of action, the Neurospora crassa assay showed a blotchy appearance in the inhibition halo produced by this chalcone, strongly suggesting that it could act by inhibiting the fungal cell wall. This chalcone seems to be an hyphal malformation inducer, since a clear curling of the hypha was observed in this hazy zone at a magnification of x 400. This work strongly contributes to the knowledge of the antifungal properties of hydroxy-chalcones. PMID- 15799011 TI - Parenteral ziprasidone: a new atypical neuroleptic for emergency treatment of psychosis in Parkinson's disease? AB - This is a report on a case series of five patients with psychosis in Parkinson's disease who were treated successfully with an intramuscular injection of ziprasidone (10-20 mg) for acute agitation. No deterioration of motor function or other clinically relevant side effects were seen. PMID- 15799013 TI - Synthesis and benzodiazepine receptor affinity of derivatives of the new tricyclic heteroaromatic system pyrido[3',2':5,6]thiopyrano[4,3-c]pyridazin 3(2H,5H)-one. AB - Derivatives 7-13 of a new tricyclic heteroaromatic system, pyrido[3',2':5,6]thiopyrano[4,3-c]pyridazin-3(2H,5H)-one, were prepared as potential ligands at the benzodiazepine receptor, in view of their structural analogy with potent ligands such as the pyrazoloquinolines of the CGS series II, and especially with the benzothiopyrano[4,3-c]pyridazinones VI. They were obtained starting from the versatile ketones 2,3-dihydrothiopyrano[2,3-b]pyridin 4(4H)-one 1 and the corresponding 7-methyl derivative 2, via condensation with glyoxylic acid, and reaction of the intermediate acid mixtures with hydrazine or substituted phenylhydrazines. When evaluated for their binding affinity at the benzo diazepine receptor in bovine cortical membranes, the target compounds 8-13 displayed an affinity in the micromolar/submicromolar order. A hypothesis is presented to rationalize these results. PMID- 15799014 TI - MRI correlates of alien leg-like phenomenon in corticobasal degeneration. AB - We describe the clinical and neuroradiologic correlates in two patients with the clinical picture of CBD and alien leg phenomena. The MRI brain scan in both had unique focal abnormalities in the corresponding leg area of the homunculus that may be the substrate for the alien limb features. PMID- 15799016 TI - Unusual nocturnal motor restlessness in a patient with spinocerebellar ataxia 3. AB - Video-polysomnographic monitoring of a female patient with spinocerebellar ataxia 3 (SCA-3) in whom rapid eye movement (REM) sleep behavior disorder was clinically suspected demonstrated recurrent episodes of unusual nocturnal motor restlessness occurring during non-REM sleep and severely disturbing both the patient and her bed partner's sleep. Even if nocturnal manifestations are common features of SCA 3 and increasingly recognized, such a case has never been reported before. We discuss this peculiar movement disorder as a particular manifestation of the wide spectrum of parasomnias occurring in such a rare neurodegenerative disease. PMID- 15799015 TI - Standardized clinical outcome rating scale for depression for use in clinical practice. AB - The integration of research into clinical practice to conduct effectiveness studies faces multiple obstacles. One obstacle is the burden of completing research measures of outcome. A simple, reliable, and valid measure that could be rated at every visit, incorporated into a clinician's progress note, and reflect the DSM-IV definition of a major depressive episode (including partial and full remission from the episode) would enhance the ability to conduct effectiveness research. The goal of the present study was to examine the reliability and validity of such a measure. Three hundred and three psychiatric outpatients who were being treated for a DSM-IV major depressive episode were rated on the Standardized Clinical Outcome Rating for Depression (SCOR-D), 17-item Hamilton Rating Scale for Depression, Montgomery-Asberg Depression Rating Scale, and the Global Assessment of Functioning. We examined the correlation between the SCOR-D and the other measures, and conducted an analyses of variance to compare mean values on these measures for each rating point on the SCOR-D. The inter-rater reliability of the SCOR-D dimensional ratings and categorical determination of remission were high. The SCOR-D was highly correlated with the other scales, and there were significant differences on the other measures of depression severity between each adjacent rating level of the SCOR-D. The SCOR-D is a brief standardized outcome measure linked to the DSM-IV approach toward defining remission that can be incorporated into routine clinical practice without adding undue burden to the treating clinician with some evidence of reliability and validity. This measure could make it more feasible to conduct effectiveness studies in clinical practice. PMID- 15799018 TI - Single crystal and powder diffraction characterization of three polymorphic forms of Acitretin. AB - Acitretin [all-trans-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8 nonatetraenoic acid or 3-methoxy-2-methyl-17-nor-1,2,3,4-tetradehydroretinoic acid], a widely marketed oral synthetic retinoid, introduced for clinical use as effective therapy against psoriasis, was found to crystallize in three polymorphic modifications (hereafter, I, II, and III), the crystal structures of which have been determined by single-crystal diffractometry (form I) or X-ray powder diffraction methods (form II and III) from conventional laboratory data only. In these latter cases, real space techniques (simulated annealing and whole profile pattern matching) have been employed. Polymorph I crystallizes in space group P2(1), Z = 8, with unit cell parameters a = 7.894(1), b = 58.454(6), c = 8.161(1) angstroms, beta = 102.04(1) degrees, and V = 3682.9(8) angstroms3. Polymorph II crystallizes in space group P2(1)/n, Z = 4, with unit cell parameters a = 13.999(2), b = 10.714(1), c = 12.465(2) angstroms, beta = 98.76(5) degrees, and V = 1847.9(3) angstroms3. Polymorph III crystallizes in space group P2(1)/c, Z = 4, with unit cell parameters a = 3.0751(4), b = 4.0487(4), c = 14.956(2) angstroms, beta = 100.41(7) degrees, and V = 1831.3(4) angstroms3. Polymorph I, found to be identical with that deposited in the European Pharmacopeia, shows four crystallographically independent Acitretin molecules, arranged in pairs through conventional hydrogen-bonded carboxylic dimers; also in form II, carboxylic dimers are observed, located on crystallographic inversion centres, while in form III, a catameric arrangement of the carboxylic residues, winding up about the rather short monoclinic axis, generates one-dimensional chains of hydrogen-bonded Acitretin molecules. Thermal analysis showed that form I can be quantitatively transformed into form II by moderate heating near 200 degrees C, under vacuum. These results show that ab initio structural studies from conventional laboratory X-ray powder diffraction (XRPD) data are fully providing the opportunity to investigate the structural aspects of moderately complex substances also in the absence of single crystals, disclosing the crystal chemistry of a few polymorphs of pharmaceutically relevant species. PMID- 15799017 TI - Oxycodone pharmacokinetics and pharmacodynamics in the rat in the presence of the P-glycoprotein inhibitor PSC833. AB - The objective of this study was to investigate the in vivo influence of the P glycoprotein (P-gp) inhibitor PSC833 on the plasma pharmacokinetics, total brain concentrations and tail-flick latency of oxycodone in rats. Eight rats each received an infusion of PSC833 or vehicle without PSC833. One hour later, all animals received 0.3 mg/kg oxycodone as a 1-h infusion. Plasma samples were taken, and tail-flick latency was monitored during the infusion and for 2 h thereafter. The brains were collected at the end of the experiment. There were no differences between the two groups in area under the plasma oxycodone concentration-time curve from time zero to infinity, or oxycodone plasma clearance, volume of distribution at steady-state, or half-life. There were no differences in average total brain oxycodone concentrations at 180 min, nor were there any differences in average tail-flick latency for the PSC833 and control groups. In conclusion, coadministration of PSC833 did not alter the plasma pharmacokinetics, brain concentrations, or associated tail-flick latency of oxycodone, indicating that oxycodone is not a P-gp substrate in the rat. This has important clinical implications, as it indicates that oxycodone, unlike some other opioids, will not interact at the blood-brain barrier (BBB) with concomitantly administered P-gp substrates. PMID- 15799019 TI - ER Ca2+ depletion triggers apoptotic signals for endoplasmic reticulum (ER) overload response induced by overexpressed reticulon 3 (RTN3/HAP). AB - Perturbance of endoplasmic reticulum (ER) function, either by the mutant proteins not folding correctly, or by an excessive accumulation of proteins in the organelle, will lead to the unfolded protein response (UPR) or ER overload response (EOR). The signal-transducing pathways for UPR have been identified, whereas the pathway for EOR remains to be elucidated. Our previous study demonstrated that the overexpression of reticulon 3 (RTN3, also named HAP, homologue of ASY protein) caused apoptosis with the depletion of ER Ca(2+) stores. In present research, we characterized RTN3 as a novel EOR-induced protein, triggering the apoptotic signals through the release of ER Ca(2+) and the elevation of cytosolic Ca(2+). Our studies showed that overexpressed RTN3 induced EOR, eliciting ER-specific apoptosis with activation of caspase-12 and mitochondrial dysfunction through ER Ca(2+) depletion and the sustained elevation of cytosolic Ca(2+). Furthermore, we demonstrated that overexpressed RTN3 and stimuli that activate both EOR and UPR, not UPR only, were able to induce up regulation of inducible nitric oxide synthase (iNOS) in HeLa cells through ER Ca(2+) release and reactive oxygen intermediates (ROIs), resulting in endogenous calcium-dependent nitric oxide protecting cells against ER specific apoptosis, which suggested that the nitric oxide and iNOS represented a likely protective response to EOR, not the UPR. These results supported that the release of ER Ca(2+) stores triggered the initial signal-transducing pathways for EOR induced by overexpressed RTN3. PMID- 15799020 TI - Only tryptophan hydroxylase (TPH)-2 is relevant to the CNS. PMID- 15799021 TI - Trichloroethylene exposure and congenital heart defects. PMID- 15799022 TI - Go G-proteins mediate rapid heterologous desensitization of G-protein coupled receptors in Xenopus oocytes. AB - We have shown previously that responses to lysophosphatidic acid (LPA) in Xenopus oocytes exhibit pronounced rapid homologous desensitization mediated by Go family of G-proteins (Itzhaki-Van Ham et al., 2004, J Cell Physiol, 200: 125-133). The present study was aimed at examining the involvement of Go G-proteins in rapid heterologous desensitization of native and expressed G-protein-coupled receptors in Xenopus oocytes. Threshold stimulation of the native lysophosphatidic acid receptors (LPA-Rs) induced about 50% rapid desensitization of responses evoked by stimulation of either native trypsin or expressed M1-muscarinic cholinergic receptors (M1-Rs). Similarly, threshold stimulation of expressed M1-Rs or thyrotropin-releasing hormone receptors induced 40% rapid desensitization of responses to LPA. Inactivation of all Gi/o G-proteins with pertussis toxin (PTX) completely abolished rapid heterologous desensitization in all protocols. Depletion of either Galphao or Galphao1 by antisense oligodeoxynucleotides targeted at either member of the Galphao family decreased or completely abolished rapid heterologous desensitization. Expression of two dominant negative mutants of the human Galphao family, highly homologous to oocyte Galphao species, either decreased or virtually abolished rapid desensitization. Homologous and heterologous desensitizations of the LPA response were non-additive and proceeded, apparently, via the same pathway. We conclude that Go G-proteins mediate both homologous and heterologous rapid desensitization of responses mediated by G-protein-coupled receptors (GPCRs) coupled to the phosphoinositide phospholipase C-inositol 1,4,5-trisphosphate-Ca(2+) (PI-PLC-InsP(3)-Ca(2+)) pathway in Xenopus oocytes. PMID- 15799023 TI - Transforming growth factor-beta1 signaling participates in the physiological and pathological regulation of mouse inner ear development by all-trans retinoic acid. AB - BACKGROUND: Retinoic acid (RA) is a vitamin A derivative that participates in patterning and regulation of inner ear development. Either excess RA or RA deficiency during a critical stage of inner ear development can produce teratogenic effects. Previous studies have shown that in utero exposure of the developing mouse inner ear to a high dose of all-trans RA (atRA) results in severe malformations of the inner ear that are associated with diminished levels of endogenous transforming growth factor-beta1 (TGF-beta(1)) protein. METHODS: In this study, the effects of a teratogenic level of atRA on levels and patterns of expression of TGFbeta receptor II (TGFbetaRII) and Smad2, a downstream component of the TGFbeta signal transduction pathway, are investigated in the developing mouse inner ear. The expression pattern of endogenous RA receptor alpha (RARalpha) and the ability of an RARalpha(1)-specific antisense oligonucleotide (AS) to modulate otic capsule chondrogenesis are demonstrated in the inner ear and in culture. RESULTS: Endogenous TGFbetaRII and Smad2 are downregulated in the inner ear following in utero atRA treatment. In addition, a reduction in endogenous TGFbeta(1) and a marked suppression of chondrogenesis occur in RARalpha(1) AS-treated cultures in comparison to untreated or oligonucleotide treated control cultures. This chondrogenic suppression can be partially overcome by supplementation of RARalpha(1) AS-treated cultures with exogenous TGFbeta(1) protein. CONCLUSIONS: Our findings support a role for TGFbeta in the physiological and pathological effects of RA on inner ear development. PMID- 15799025 TI - Reduced folate carrier gene is a risk factor for neural tube defects in a Chinese population. AB - BACKGROUND: There is a considerable body of data demonstrating that periconceptional supplementation of folic acid can prevent a significant proportion of neural tube defects (NTDs). At present, the mechanism by which folic acid exerts its beneficial effect remains unknown. Folate transporter genes, including the reduced folate carrier gene (RFC1), have been proposed as NTD risk factors. METHODS: The study population included 104 nuclear families with NTDs and 100 nonmalformed control families. We investigated the possible association between a common RFC1 polymorphism (A80G) and NTD risk among offspring, as well as potential gene-environment interactions between the infant RFC1 genotype and maternal periconceptional use of folic acid through a population-based case-control study. RESULTS: We observed that the infants of the GG genotype were associated with a 2.56-fold increased risk of NTDs when compared to the AA genotype (odds ratio [OR], 2.56; 95% confidence interval [CI], 1.04 6.36) in our study population. Among mothers who did not utilize folic acid supplements, the risk for having a child with an NTD was 3.30 (95% CI, 1.15-9.65) for offspring with the GG genotype, compared to the reference (AA) genotype. Children who had the GG genotype and whose mothers did not take folic acid had an elevated risk for NTDs (OR, 8.80; 95% CI, 2.83-28.69), compared to offspring with the AA and GA genotypes whose mothers utilized folic acid supplements. CONCLUSIONS: Our findings suggest that the RFC1 G allele is likely to be an important genetic factor in determining folate transport and subsequently may be a risk factor for NTDs in this Chinese population. PMID- 15799026 TI - Identification of early-responsive genes correlated to valproic acid-induced neural tube defects in mice. AB - BACKGROUND: Valproic acid (VPA) causes the failure of neural tube closure in newborn mice. However, the molecular mechanism of its teratogenesis is unknown. This study was conducted to investigate the genomewide effects of VPA disruption of normal neural tube development in mice. METHODS: Microarray analysis was performed on the head part of NMRI mouse embryos treated for 1 hr with VPA on gestational day (GD) 8. Subsequently, we attempted to isolate genes that changed in correlation with the teratogenic action of VPA by employing reduced teratogenic VPA analogs, valpromide (VPD) and valnoctamide (VCD), in a real-time PCR study. RESULTS: Microarray results demonstrated that during neurulation, many genes, some of whose functions are known and some unknown, were either increased or decreased after VPA injection. Some genes were affected by VPD or VCD in the same way as VPA, but others were not changed by the analogs. In this way, our system identified 11 increased and 20 decreased genes. Annotation analysis revealed that the increased genes included gadd45b, ier5, per1, phfl3, pou3f1, and sox4, and the decreased genes included ccne2, ccnl, gas5, egr2, sirt1, and zfp105. CONCLUSIONS: These findings demonstrate that expression changes in genes having roles in the cell cycle and apoptosis pathways of neural tube cells were strongly expected to relate to the teratogenic, but not antiepileptic, activity of VPA. Our approach has allowed the expansion of the catalog of molecules immediately affected by VPA in the developing neural tube. PMID- 15799027 TI - Cooperation between antioxidants and 1,25-dihydroxyvitamin D3 in induction of leukemia HL60 cell differentiation through the JNK/AP-1/Egr-1 pathway. AB - Vitamin D derivatives have demonstrated anti-cancer activity, but their clinical use is precluded by hypercalcemia. Previously, we found that carnosic acid potentiates differentiation of human leukemia cells induced by low concentrations of 1alpha,25-dihydroxyvitamin D(3) (1,25D(3)). In this study, we investigated if this effect is a general property of antioxidants, and whether there is a common mechanism whereby antioxidants potentiate monocytic differentiation. We found that all antioxidants tested enhanced differentiation-related cell cycle arrest induced by a low (1 nM) concentration of 1,25D(3). Addition of antioxidants to 1,25D(3) activated the JNK pathway as indicated by increased phosphorylation of c jun and ATF-2, although each compound alone had a minimal effect. Antioxidants also enhanced the 1,25D(3)-induced AP-1 DNA binding and transactivation ability. Expression of Egr-1 and c-fos was increased by combinations of antioxidants and 1,25D(3), in parallel with the activation of the JNK pathway. The potentiation of differentiation by antioxidants was inhibited by JNK inhibitor SP600125 and a dominant negative JNK 1/2 construct, and Egr-1 and c-fos expression was proportionally decreased, suggesting that JNK pathway regulates these transcription factors. While potentiating the prodifferentiation effect of 1,25D(3), antioxidants did not promote the elevation of basal levels of intracellular calcium by 1,25D(3). The results indicate that JNK-AP1 pathway has an important role in the potentiation of 1,25D(3)-induced differentiation by antioxidants, and regulates expression of Egr-1 and c-fos. Combinations of antioxidants with 1,25D(3) should be further evaluated for use in cancer chemoprevention and therapy. PMID- 15799028 TI - Teratogenic effects of retinoic acid are modulated in mice lacking expression of epidermal growth factor and transforming growth factor-alpha. AB - BACKGROUND: Epidermal growth factor (EGF) and transforming growth factor-alpha (TGFalpha) regulate cell proliferation and differentiation in the embryo. The induction of cleft palate (CP) by all trans-retinoic acid (RA) was associated with altered expression of TGFalpha, EGF receptor, and binding of EGF. This study uses knockout (KO) mice to examine the roles of EGF and TGFalpha in teratogenic responses of embryos exposed to RA. METHODS: Pregnant wild-type (WT) mice of mixed genetic background, EGF KO, C57BL/6J, and TGFalpha KO mice were given a single oral dose of RA (100 mg/kg, 10 ml/kg) or corn oil on GD 10 at 12 PM, GD 11 at 12 PM or 4 PM, or GD 12 at 8 AM or 12 PM (plug day = GD 0). GD 18 fetuses were examined for external, visceral, and skeletal effects. RESULTS: After exposure to RA on GD 12, the incidence of CP in EGF KO was significantly reduced relative to WT. In TGFalpha KO fetuses, RA exposure on GD 10 increased the incidence of CP versus C57BL/6J. The incidence of skeletal defects in the limbs, vertebrae, sternebrae, and ribs were also affected by lack of expression of EGF or TGFalpha with region-specific amelioration or exacerbation of the effects of RA. In TGFalpha KO fetuses, incidences of forelimb long bone and digit defects increased relative to C57BL/6J. In EGF KO fetuses, relative to WT, the incidence of hindlimb oligodactyly was increased. In EGF KO, but not WT, RA produced short, bent radius, humerus, and ulna. Both TGFalpha and EGF KO mice had increased incidences of dilation of the renal pelvis and this was reduced by RA. CONCLUSIONS: RA exposure produced skeletal and visceral defects in all genotypes; however, EGF or TGFalpha KO influenced the incidence and severity of defects. This study supports a role for EGF and TGFalpha in the response to RA. PMID- 15799029 TI - Osteoprotegerin (OPG) is localized to the Weibel-Palade bodies of human vascular endothelial cells and is physically associated with von Willebrand factor. AB - Recent studies demonstrate roles for osteoprotegerin (OPG) in both skeletal and extra-skeletal tissues. Although its role in preventing osteoclast (OC) formation and activity is well documented, emerging evidence suggests a role of OPG in endothelial cell survival and the prevention of arterial calcification. In this communication, we show that vascular endothelial cells in situ, and human umbilical vein endothelial cells (HUVEC) in vitro, express abundant OPG. In HUVEC, OPG co-localizes with P-selectin and von Willebrand factor (vWF), within the Weibel-Palade bodies (WPB). Treatment of HUVEC with the pro-inflammatory cytokines, tumor necrosis factor (TNF)-alpha and IL-1beta, resulted in mobilization from the WPBs and subsequent secretion of OPG protein into the culture supernatant. Furthermore, TNF-alpha treatment of HUVEC resulted in a sustained increase in OPG mRNA levels and protein secretion over the 24-h treatment period. Reciprocal immunoprecipitation experiments revealed that while not associated with P-Selectin, OPG is physically complexed with vWF both within the WPB and following secretion from endothelial cells. Interestingly, this association was also identified in human peripheral blood plasma. In addition to its interaction with vWF, we show that OPG also binds with high avidity to the vWF reductase, thrombospondin (TSP-1), raising the intriguing possibility that OPG may provide a link between TSP-1 and vWF. In summary, the intracellular localization of OPG in HUVEC, in association with vWF, together with its rapid and sustained secretory response to inflammatory stimuli, strongly support a modulatory role in vascular injury, inflammation and hemostasis. PMID- 15799030 TI - Effects of amphotericin B on ion transport proteins in airway epithelial cells. AB - Topical intranasal application of the antifungal Amphotericin B (AmphoB) has been shown as an effective medical treatment of chronic rhinosinusitis. Because this antibiotic forms channels in lipid membranes, we considered the possibility that it affects the properties and/or cell surface expression of ion channels/pumps, and consequently transepithelial ion transport. Human nasal epithelial cells were exposed apically to AmphoB (50 microM) for 4 h, 5 days (4 h daily), and 4 weeks (4 h daily, 5 days weekly) and allowed to recover for 18-48 h. AmphoB significantly reduced transepithelial potential difference, short-circuit current, and the amiloride-sensitive current. This was not due to generalized cellular toxicity as judged from normal transepithelial resistance and mitochondrial activity, but was related to inhibitory effects of AmphoB on ion transport proteins. Thus, cells exposed to AmphoB for 4 h showed decreased apical epithelial sodium channels (ENaC) activity with no change in basolateral Na(+)K(+)-ATPase activity and K(+) conductance, and reduced amount of alphaENaC, alpha1-Na(+)K(+)-ATPase, and NKCC1 proteins at the cell membrane, but no change in mRNA levels. After a 5-day treatment, there was a significant decrease in Na(+)K(+)-ATPase activity. After a 4-week treatment, a decrease in basolateral K(+) conductance and in alphaENaC and alpha1-Na(+)K(+)-ATPase mRNA levels was also observed. These findings may reflect a feedback mechanism aimed to limit cellular Na(+) overload and K(+) depletion subsequently to formation of AmphoB pores in the cell membrane. Thus, the decreased Na(+) absorption induced by AmphoB resulted from reduced cell surface expression of the ENaC, Na(+)K(+) ATPase pump and NKCC1 and not from direct inhibition of their activities. PMID- 15799031 TI - Chondrocyte cluster formation in agarose cultures as a functional assay to identify genes expressed in osteoarthritis. AB - Understanding altered gene expression in osteoarthritic cartilage can lead to new targets for drug intervention. We established a functional assay based on chondrocyte cluster formation, a phenotype associated with osteoarthritis (OA), to screen an OA cartilage gene library. Previous reports have demonstrated that normal chondrocytes grown in suspension culture maintain their chondrocytic phenotype, however, certain growth factors such as basic fibroblast growth factor (bFGF) will induce the cells to proliferate in tight clusters similar to those seen in osteoarthritic cartilage. In this study we validate that overexpression of bFGF by retrovirally transduced normal chondrocytes would similarly induce the proliferation of tight cell clusters. We then used this approach as a basis to set up a functional screen where an entire OA cartilage cDNA library was tranduced into normal chondrocytes to search for other genes that would also induce cluster formation. Seven potential genes were isolated from the OA gene library, including BPOZ, IL-17 receptor C, NADH ubiquinone oxidoreductase, COMP, Soluble carrier 16 (MCT 3), C1r, and bFGF itself. None of the identified genes were upregulated by bFGF, however, all of them upregulated the expression of bFGF suggesting a common pathway. Although cluster formation is not considered to be destructive in OA cartilage, it is consistent with the disease and could yield answers to the altered phenotype. Further studies are needed to elucidate how these genes are linked to the disease state. PMID- 15799032 TI - Differential regulation of rodent hepatocyte and oval cell proliferation by interferon gamma. AB - Hepatocytes and intrahepatic progenitor cells (oval cells) have similar responses to most growth factors but rarely proliferate together. Oval cells constitute a reserve compartment that is activated when hepatocyte proliferation is inhibited. Interferon gamma (IFN-gamma) increases in liver injury that involves oval cell responses, but it is not upregulated during liver regeneration after partial hepatectomy. Based on these observations, we used well-characterized lines of hepatocytes (AML-12 cells) and oval cells (LE-6 cells) to investigate the potential mechanisms that regulate differential growth responses in hepatocytes and oval cells. We show that IFN-gamma blocks hepatocyte proliferation in vivo, and that in combination with either tumor necrosis factor (TNF) or lipopolysaccharide (LPS), it causes cell cycle arrest in hepatocytes but stimulates oval cell proliferation in cultured cells. The hepatocyte cell cycle arrest is reversible, is p53-independent, and is not associated with apoptosis. Treatment of AML-12 hepatocytes with IFN-gamma/LPS or IFN-gamma/TNF, but not with individual cytokines, induced NO synthase and generated NO, while similarly treated oval cells produced little if any NO. Generation of NO by an NO donor reproduced the inhibitory effect of the cytokine combinations on AML-12 cell replication, while NO inhibitors abolish the replication deficiency. In conclusion, we propose that IFN-gamma, in conjunction with TNF or LPS, can both inhibit hepatocyte proliferation through the generation of NO and stimulate oval cell replication. The response of hepatocytes and oval cells to cytokine combinations may contribute to the differential proliferation of these cells in hepatic growth processes. PMID- 15799033 TI - Adenoviral infection of survivin antisense sensitizes prostate cancer cells to etoposide in vivo. AB - BACKGROUND: We aimed to investigate whether use of a survivin antisense fragment carried by an adenovirus vector (Ad.survivin-AS) could enhance the therapeutic efficacy of chemotherapy for androgen-independent prostate cancer. METHODS: We used Ad.survivin-AS to promote apoptosis through inhibition of survivin expression. Recombinant adenoviruses alone or in combination with chemotherapeutic agents were tested for anti-cancer activity both in vitro and in vivo. RESULTS: Infection with Ad.survivin-AS strongly inhibited survivin expression in a dose- and time-dependent manner, resulting in significant antitumor activity in vitro and in vivo. Downregulation of survivin expression potentiated induction of apoptosis by the chemotherapeutic agents docetaxel and etoposide in DU145 cells. In particular, the combination of etoposide and Ad.survivin-AS demonstrated dramatic growth inhibition with no tumor regrowth being observed during the experimental period. CONCLUSIONS: The prominent synergy of this combination may provide a basis for clinical application of Ad.survivin AS as a chemosensitizer of etoposide. PMID- 15799034 TI - Mechanisms of benzarone and benzbromarone-induced hepatic toxicity. AB - Treatment with benzarone or benzbromarone can be associated with hepatic injury. Both drugs share structural similarities with amiodarone, a well-known mitochondrial toxin. Therefore, we investigated the hepatotoxicity of benzarone and benzbromarone as well as the analogues benzofuran and 2-butylbenzofuran. In isolated rat hepatocytes, amiodarone, benzarone, and benzbromarone (20 micromol/L) decreased mitochondrial membrane potential by 23%, 54% or 81%, respectively. Benzofuran and 2-butylbenzofuran had no effect up to 100 micromol/L. In isolated rat liver mitochondria, amiodarone, benzarone, and benzbromarone, but not benzofuran, decreased state 3 oxidation and respiratory control ratios for L-glutamate (50% decrease of respiratory control ratio at [micromol/L]: amiodarone, 12.9; benzarone, 10.8; benzbromarone, <1). Amiodarone, benzarone, and benzbromarone, but not benzofuran, also uncoupled oxidative phosphorylation. Mitochondrial beta-oxidation was decreased by 71%, 87%, and 58% with 100 micromol/L amiodarone or benzarone and 50 micromol/L benzbromarone, respectively, but was unaffected by benzofuran, whereas ketogenesis was not affected. 2-Butylbenzofuran weakly inhibited state 3 oxidation and beta-oxidation only at 100 micromol/L. In the presence of 100 micromol/L amiodarone, benzarone or benzbromarone, reactive oxygen species production was increased, mitochondrial leakage of cytochrome c was induced in HepG2 cells, and permeability transition was induced in isolated rat liver mitochondria. At the same concentrations, amiodarone, benzarone, and benzbromarone induced apoptosis and necrosis of isolated rat hepatocytes. In conclusion, hepatotoxicity associated with amiodarone, benzarone, and benzbromarone can at least in part be explained by their mitochondrial toxicity and the subsequent induction of apoptosis and necrosis. Side chains attached to the furan moiety are necessary for rendering benzofuran hepatotoxic. PMID- 15799035 TI - Fast, accurate, and automatic extraction of the modified Talairach cortical landmarks from magnetic resonance images. AB - The Talairach transformation is the most prevalent way to normalize brains and is hindered by, among others things, a lack of automatic determination of cortical landmarks. An algorithm to locate the modified Talairach cortical landmarks in three steps is proposed: determination of the three planes containing the landmarks; segmentation of the planes based on range-constrained thresholding and morphologic operations; and local refinement of the segmentation to locate the landmarks. The algorithm has been validated against 62 T(1)-weighted and SPGR MR diversified data sets. For each data set, it takes less than 2 s on a Pentium 4 to extract all six landmarks. The average landmark location errors are below 0.9 mm. The algorithm is robust due to incorporation of anatomic knowledge. A low computational cost results from processing of three 2D images and employing only simple operations like thresholding, basic morphologic operations, and distance transform. PMID- 15799036 TI - Short breath-hold, volumetric coronary MR angiography employing steady-state free precession in conjunction with parallel imaging. AB - An ECG-gated, 3D steady-state free precession (SSFP) technique in conjunction with sensitivity encoding (SENSE)-based parallel imaging was implemented for short breath-hold, volumetric coronary MR angiograpy (CMRA). Two parallel imaging acquisition strategies (employing 1 R-R and 2 R-R intervals, respectively) were developed to achieve 1) very short breath-hold times (12 s for a heart rate of 60 bpm), and 2) small acquisition windows to minimize sensitivity to physiologic motion. Both strategies were examined in CMRA applications over a range of heart rates. A four-point scale blinded reading (with 4 indicating the most desirable features) revealed substantial image quality improvements for the accelerated data as compared to the nonaccelerated approach. The 1 R-R interval scheme yielded an image score of 3.39 +/- 0.60, and was found to be particularly suitable for low heart rates (P = 0.0008). The 2 R-R interval strategy yielded an image score of 3.35 +/- 0.64, and was more appropriate for higher heart rates (P = 0.03). The results demonstrate that 3D SSFP combined with parallel imaging is a versatile method for short breath-hold CMRA while maintaining high spatial resolution. This strategy permits imaging of the major coronary artery distributions in two to three breath-holds using targeted slabs, and offers the potential for single breath-hold, large-volume CMRA. PMID- 15799037 TI - Contrast-to-noise ratios of diffusion anisotropy indices. AB - The degree of diffusion anisotropy in different brain regions is usually measured by a diffusion anisotropy index (DAI) such as relative anisotropy (RA) and fractional anisotropy (FA). FA has been reported to have a higher contrast-to noise ratio (CNR) than RA. The present work compares the CNRs of seven DAIs in theoretical propagation-of-error calculations, in simulations, and in human brain measurements over small and large anisotropy differences. In simulations all seven CNRs were similar for small anisotropy differences. Small differences among the DAIs appeared at higher anisotropy levels and lower signal-to-noise ratios with certain tensor orientations. The DAIs fell into three groups based on algebraic relationships and small CNR differences. The group with RA and FA had the best CNR. Human brain regions with small anisotropy differences had similar CNR for all seven DAIs, and the scatter in the data was greater than any expected differences. With large anisotropy differences, a small advantage appeared for RA over FA in some simulations and for FA over RA in other simulations. The CNR between brain regions with very different anisotropies was different for each DAI. The apparent reported advantage of FA over RA is explained by biologic heterogeneity and by noise-induced bias in the DAI values and their standard deviations. PMID- 15799038 TI - Contrast-enhanced MRI with new biodegradable macromolecular Gd(III) complexes in tumor-bearing mice. AB - The structures of polydisulfide-based biodegradable macromolecular Gd(III) complexes were modified to improve their in vivo retention time and MRI contrast enhancement. Steric hindrance was introduced around the disulfide bonds to control their access to free thiols in order to alter the degradation rate of the copolymers. Two new macromolecular agents, (Gd-DTPA)-cystine copolymers (GDCP) and (Gd-DTPA)-cystine diethyl ester copolymers (GDCEP), were prepared. Both agents were readily degraded in vitro and in vivo by the disulfide-thiol exchange reaction, but at a slow rate. The introduction of COOH and COOEt groups slowed down the degradation of the copolymers in the incubation with 15 microM cysteine. Metabolic degradation products were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry in the urine samples from rats injected with the agents. The T(1) relaxivity (r(1)) was 5.43 mM(-1)s(-1) for GDCP, and 5.86 mM(-1)s(-1) for GDCEP, respectively, at 3T. MRI contrast enhancement of both agents was studied in nude mice bearing MDA-BM-231 human breast carcinoma xenografts, on a Siemens Trio 3T scanner. The modified agents resulted in more significant contrast enhancement in the blood pool and tumor periphery than (Gd-DTPA)-cystamine copolymers (GDCC) and a low-molecular weight control agent, Gd-(DTPA-BMA), at a dose of 0.1 mmol-Gd/kg. The results demonstrate that the structural modification of the biodegradable macromolecular Gd(III) complexes resulted in a relatively slow degradation of the macromolecules and significantly improved in vivo contrast enhancement. The modified agents show promise for use in investigations of blood pool and cancer by contrast-enhanced (CE) MRI. PMID- 15799039 TI - Generalized scalar measures for diffusion MRI using trace, variance, and entropy. AB - This paper details the derivation of rotationally invariant scalar measures from higher-rank diffusion tensors (DTs) and functions defined on a unit sphere. This was accomplished with the use of an expression that generalizes the evaluation of the trace operator to tensors of arbitrary rank, and even to functions whose domains are the unit sphere. It is shown that the mean diffusivity is invariant to the selection of tensor rank for the model used. However, this rank invariance does not apply to the anisotropy measures. Therefore, a variance-based, general anisotropy measure is proposed. Also an information theoretical parametrization of anisotropy is introduced that is frequently more consistent with the meaning attributed to anisotropy. We accomplished this by associating anisotropy with the amount of orientational information present in the data, regardless of the imaging technique used. Using a simplified model of fibrous tissue, we simulated anisotropy values with varying orientational complexity and tensor models. Simulations suggested that a lower-rank tensor model may produce artificially low anisotropy values in voxels with complex structure. This was confirmed with a spin-echo experiment performed on an excised rat brain. PMID- 15799040 TI - Visualization of cortical spreading depression using manganese-enhanced magnetic resonance imaging. AB - Cortical spreading depression (CSD) was visualized using manganese-enhanced MRI (MEMRI) following topical application of KCl to the exposed rat cortex. MEMRI signal increase in the ipsilateral cortex relative to the contralateral control region was 60 +/- 30% following two KCl applications. MEMRI signal increase for a single (40%) versus double (80%) KCl application suggests an integration effect over successive CSD episodes. CSD-induced MEMRI enhancement involved cortical layers containing dense regions of apical dendrites, supporting the contention that these neuronal structures are necessary for propagation of CSDs. Subcortical enhancement was present in hippocampal and thalamic regions, most likely a result of neuronal connections with cortical layers 4 and 5. These results are consistent with previous studies of CSD using diffusion-weighted MRI and T(2) (*) weighted MRI and should be useful for investigating CSD itself and its role in other neurologic disorders. PMID- 15799041 TI - 1H MR spectroscopy using TE averaged PRESS: a more sensitive technique to detect neurodegeneration associated with Alzheimer's disease. AB - A sensitive proton magnetic resonance spectroscopy ((1)H MRS) acquisition scheme that is capable of discriminating between normal controls and a group of patients with early Alzheimer's disease (AD) is presented. The performance of this newly developed method, TE averaged PRESS (PRESS-J), in detecting neurodegeneration associated with early AD is compared with that of short echo time (TE) PRESS. A stepwise discriminant function analysis is used to construct discriminant functions for both pulse sequences. These functions are each composed of a single predictor: the N-acetyl aspartate (NAA)/creatine (Cr) ratio for PRESS-J, and the NAA/myoInositol (mI) ratio for PRESS. We observed lower P-values, higher areas under the receiver operating characteristic curves, and higher sensitivity at a given specificity for the PRESS-J discriminating function in comparison with the PRESS discriminating function. The higher sensitivity of PRESS-J is due to decreased variability when the singlets are fit in the spectra. This increased sensitivity enables new MR applications and, among other benefits, allows for smaller group sizes in drug trials, which can significantly reduce the cost of such trials. PMID- 15799042 TI - Variability of metabolite yield using STEAM or PRESS sequences in vivo at 3.0 T, illustrated with myo-inositol. AB - Using as an example the myo-inositol (mI) band at approximately 3.6 ppm in the proton spectrum from brain, an evaluation is presented that highlights the difficulties of quantifying metabolites with strongly coupled spins with either STEAM or PRESS and demonstrates some advantages of prospective sequence analysis when measuring their concentrations. The analysis emphasizes the variation in coupled-spin signal yield and lineshape, compared with that of uncoupled singlets such as N-acetylaspartate, a variation that differs from one metabolite spin system to another. This difference in variation between a target metabolite (e.g., mI) and its contaminating background metabolites (e.g., glutamate and taurine, etc.) is shown to provide in certain circumstances a substantial reduction in background contamination (both metabolite and macromolecule) while maintaining sufficient signal-to-noise ratio for precise quantification. For example, sequence times are demonstrated, both for STEAM and for PRESS, that, relative to the short echo-time sequences typical in the literature, enhance the signal to metabolite background of the 3.6-ppm band of mI by factors of 1.7 and 1.3, respectively, essentially eliminate the macromolecular baseline, and yet in vivo retain an S/N approximately 10 in both cases. PMID- 15799043 TI - Iopamidol: Exploring the potential use of a well-established x-ray contrast agent for MRI. AB - Iopamidol is one of the most common contrast media used for diagnostic CT-based clinical protocols. Chemically, this molecule contains two pools of mobile protons (amide and alcoholic) that are in exchange with water. At 7.05 T, pH 7.4, and 312 K, the exchange rate of the alcoholic protons is too fast to affect the NMR properties of water protons, whereas the slowly exchanging amide protons induce a T(2)-shortening effect on the "bulk" water signal that is detectable when the concentration is about 12 mM. Moreover, a more pronounced contrast is observed when the amide resonances are saturated by the application of an appropriate RF irradiation field, making iopamidol a potential chemical exchange saturation transfer (CEST) agent whose effect can be detected at a concentration as low as 7 mM (at 7.05 T). The exploitation of the MRI properties of iopamidol could facilitate novel and interesting diagnostic applications for combined MRI and CT studies. PMID- 15799044 TI - Dynamic autocalibrated parallel imaging using temporal GRAPPA (TGRAPPA). AB - Current parallel imaging techniques for accelerated imaging require a fully encoded reference data set to estimate the spatial coil sensitivity information needed for reconstruction. In dynamic parallel imaging a time-interleaved acquisition scheme can be used, which eliminates the need for separately acquiring additional reference data, since the signal from directly adjacent time frames can be merged to build a set of fully encoded full-resolution reference data for coil calibration. In this work, we demonstrate that a time-interleaved sampling scheme, in combination with autocalibrated GRAPPA (referred to as TGRAPPA), allows one to easily update the coil weights for the GRAPPA algorithm dynamically, thereby improving the acquisition efficiency. This method may update coil sensitivity estimates frame by frame, thereby tracking changes in relative coil sensitivities that may occur during the data acquisition. PMID- 15799045 TI - 3He lung imaging in an open access, very-low-field human magnetic resonance imaging system. AB - The human lung and its functions are extremely sensitive to gravity; however, the conventional high-field magnets used for most laser-polarized (3)He MRI of the human lung restrict subjects to lying horizontally. Imaging of human lungs using inhaled laser-polarized (3)He gas is demonstrated in an open-access very-low magnetic-field (<5 mT) MRI instrument. This prototype device employs a simple, low-cost electromagnet, with an open geometry that allows variation of the orientation of the imaging subject in a two-dimensional plane. As a demonstration, two-dimensional lung images were acquired with 4-mm in-plane resolution from a subject in two orientations: lying supine and sitting in a vertical position with one arm raised. Experience with this prototype device will guide optimization of a second-generation very-low-field imager to enable studies of human pulmonary physiology as a function of subject orientation. PMID- 15799046 TI - Adrenergic stimulation of Rb+ uptake in pig hearts in vivo assessed by 87Rb MRS. AB - It was previously shown that the adrenergic agonist dobutamine stimulates Rb(+) uptake in isolated pig hearts. In the present work we examined whether dobutamine can increase Rb(+) uptake rate in vivo. Open-chest domestic pigs (N = 14) were used under general anesthesia. The surface coil was placed against the anterior left ventricular wall to obtain (87)Rb spectra using a spectrometer interfaced with the 7T, 30-cm horizontal bore magnet. RbCl was infused at the rate of 1.35 +/- 0.14 mmol/kg/hr without or with dobutamine (0.6 mg/kg/hr) over a 60-min period, and then the infusions were terminated. The rate constant (k x 10(3), from 13 +/- 2.4 to 36 +/- 12 min(-1)) and Rb(+) influx rate (from 2.5% to 4.8%/min) were increased by dobutamine despite lower plasma [Rb(+)] (0.59 +/- 0.16 vs. 0.84 +/- 0.24 mM in control), while the tissue/plasma Rb ratios were identical (38 +/- 9). Heart rate (HR) and systolic blood pressure (SBP) were increased from 106 +/- 9 to 161 +/- 15 bpm (+52%) and from 78 +/- 7 to 93 +/- 11 mmHg (+19%), respectively. The stimulation of Rb(+) uptake by dobutamine is consistent with the activation of Na(+)/K(+) ATPase previously observed in isolated hearts. However, the 50% increase in HR and the double coronary flow caused by the SBP increase and vasodilatation may also contribute to this effect. PMID- 15799048 TI - Real-time magnetic resonance with physiologic monitoring for improved scan localization. AB - Imaging of the coronary arteries at diagnostic resolutions is made difficult due to cardiac and respiratory motion during data acquisition. Cardiac gating and respiratory gating or breath holding are effective ways to reduce the effects of motion. The optimal cardiac and respiratory timings vary widely across individuals. This work presents a real-time magnetic resonance imaging approach with physiologic monitoring that can be used to predict the optimal timings on a subject-by-subject basis during a brief real-time prescan. The feasibility of this approach at determining the optimal cardiac trigger delay and respiratory phase is demonstrated. PMID- 15799047 TI - Dynamic dephasing changes in developing ischemic cerebral infarction in rats studied by Carr-Purcell T2 magnetic resonance imaging. AB - Carr-Purcell (CP) T(2) MRI with adiabatic pulses, acquired with varying interecho interval (tau(CP)), was used to study the time course of T(2) and relative dynamic-dephasing contrast in the rat brain. Exposure to 30 min of middle cerebral artery occlusion (MCAo) resulted in an irreversible increase in absolute CP-T(2) relaxation times. This was not associated with signal change in the relative dynamic-dephasing images, as computed by subtracting short tau(CP) CP T(2) images from long tau(CP) images and normalizing for long tau(CP) images. A day after MCAo strong CP-T(2) hyperintensity and low apparent diffusion coefficient were evident in the striatum with a decline in relative dynamic dephasing contrast. Low dynamic dephasing contrast prevailed in striatum until day 5 post-MCAo, returning to control levels with similar time course to normalizing T(2) and diffusion. The present results show a novel behavior of dynamic-dephasing contrast in poststroke brain tissue, providing data to assess the age of infarction in association to T(2) images. PMID- 15799049 TI - Quantification of choline compounds in human hepatic tumors by proton MR spectroscopy at 3 T. AB - The quantification of choline-containing compounds (Cho) in hepatic tumors by (1)H MR spectroscopy (MRS) is of great interest because such compounds have been linked to malignancy. In this study, a practical external phantom replacement method for the absolute quantification of hepatic metabolites is demonstrated. We performed experiments at 3 T using a body coil, and used an external phantom containing choline chloride for calibration. We first tested the quantification strategy to confirm its suitability in vivo using a phantom of known concentration and normal brain tissue. The results obtained after coil loading and T(1) and T(2) effects were corrected for were consistent with the known concentration and previously published values. To demonstrate its feasibility, we applied the technique to liver studies conducted on five normal volunteers and four patients with hepatocellular carcinoma, and one patient (also in the latter group) who had undergone post-transcatheter arterial chemoembolization (TACE). The Cho concentrations in the four patients were estimated to be 3.4, 6.3, 7.4, and 14.0 mM, respectively. These values are substantially higher than those obtained from the healthy volunteers (1.3 +/- 0.9 mM (mean +/- SD)). The results indicate that the proposed method is accurate and requires fewer tedious procedures for MRS; therefore, it may be a promising technique for evaluating response to treatment in liver cancer. PMID- 15799050 TI - Input parameter sensitivity analysis and comparison of quantification models for continuous arterial spin labeling. AB - The regional cerebral blood flow (rCBF) values determined using continuous arterial spin labeling (CASL) are subject to several sources of variability, including natural physiologic variations, sensitivity to the input parameters, and the use of different quantification models. To date, a thorough analysis of the impact of input parameters and the choice of quantification model has not been performed. These sources of variability were investigated through computer simulations using bootstrap techniques on actual CASL data. Coefficients of variation for representative single voxels were 6.7% for gray matter and 29% for white matter, and for eight-voxel regions of interest they were 4.5% for gray matter and 23% for white matter. Comparison of nine CASL quantification models showed differences in gray matter rCBF values of up to 42%. An analysis of the sensitivity of the rCBF to input parameters for each of the nine quantification models demonstrated that accurate quantification of the inversion efficiency, tissue and arterial blood longitudinal relaxation times, and transit times were critical in calculating precise rCBF values. The large potential variations in rCBF and the effect of the choice of quantification model suggest that interpreting absolute rCBF values in CASL studies can be challenging and requires great care. PMID- 15799051 TI - Open half-volume quadrature transverse electromagnetic coil for high-field magnetic resonance imaging. AB - A half-volume quadrature head transverse electromagnetic (TEM) coil has been constructed for 4 T imaging applications. This coil produces a sufficiently large homogeneous B(1) field region for the use as a volume coil. It provides superior transmission efficiency, resulting in significantly lower power deposition, as well as greater sensitivity and improved patient comfort and accessibility compared with conventional full-volume coils. Additionally, this coil suppresses the RF penetration artifact that distorts the RF magnetic field profile and alters the intensity in high-field images recorded with linear surface and volume coils. These advantages make it possible to apply this device as an efficient transmit/receive coil for high-field imaging with a restricted field of view. PMID- 15799052 TI - 23Na MRI combined with contrast-enhanced 1H MRI provides in vivo characterization of infarct healing. AB - Although (23)Na MRI has been shown to delineate acute myocardial infarction (MI), the time course of in vivo (23)Na MRI during infarct healing remains unknown. In this study (23)Na MRI was combined with contrast-enhanced (CE) (1)H MRI to noninvasively characterize infarct healing in vivo. Serial in vivo 3D (23)Na MRI and (1)H MRI were performed for up to 9 weeks postinfarction in 10 dogs. Radioactive microspheres were used to measure myocardial perfusion, and Hematoxylin-Eosin (H&E) and Masson's trichrome (MT) staining were used to assess interstitial cell infiltrate and collagen content. In vivo (23)Na MRI accurately delineated infarct size up to day 5 postinfarction in comparison with (1)H MRI (8.9% +/- 8.1% vs. 8.6% +/- 7.9% on day 1 postinfarction, P = NS; and 6.3% +/- 6.2% vs. 6.2% +/- 6.2% on days 4/5 postinfarction, P = NS). The in vivo (23)Na MRI signal intensity, expressed as the signal intensity ratio of infarcted tissue vs. noninfarcted tissue (MI/R) peaked on day 1 of infarction (2.04 +/- 0.23) but decreased significantly to 1.27 at 9 weeks postinfarction (P < 0.05) due to granulation tissue infiltrate and collagen deposition. To confirm the MI/R decrease during scar formation ex vivo, we performed (23)Na MRI in 12 rats on day 3 post-MI (N = 5) and after 6 weeks (N = 7). H&E and Picrosirius Red staining confirmed granulation tissue infiltrate on day 3 and scar formation after 6 weeks. MI/R decreased significantly from 1.91 +/- 0.45 on day 3 post-MI to 1.3 +/ 0.09 after 6 weeks. Thus, in vivo (23)Na MRI accurately delineates infarct size up to day 5 postinfarction. In vivo (23)Na MRI signal intensity decreases during infarct healing as a result of the underlying infarct healing process. PMID- 15799053 TI - Effects of breathing and cardiac motion on spatial resolution in the microscopic imaging of rodents. AB - One can acquire high-resolution pulmonary and cardiac images in live rodents with MR microscopy by synchronizing the image acquisition to the breathing cycle across multiple breaths, and gating to the cardiac cycle. The precision with which one can synchronize image acquisition to the motion defines the ultimate resolution limit that can be attained in such studies. The present work was performed to evaluate how reliably the pulmonary and cardiac structures return to the same position from breath to breath and beat to beat across the prolonged period required for MR microscopy. Radiopaque beads were surgically glued to the abdominal surface of the diaphragm and on the cardiac ventricles of anesthetized, mechanically ventilated rats. We evaluated the range of motion for the beads (relative to a reference vertebral bead) using digital microradiography with two specific biological gating methods: 1) ventilation synchronous acquisition, and 2) both ventilation synchronous and cardiac-gated acquisitions. The standard deviation (SD) of the displacement was < or =100 microm, which is comparable to the resolution limit for in vivo MRI imposed by signal-to-noise ratio (SNR) constraints. With careful control of motion, its impact on resolution can be limited. This work provides the first quantitative measure of the motion-imposed resolution limits for in vivo imaging. PMID- 15799056 TI - Rotating-frame intermolecular double-quantum spin-lattice relaxation T1rho, DQC weighted magnetic resonance imaging. AB - In this study, spin-locking techniques were added as a part of intermolecular multiple-quantum experiments, thereby introducing the concept of rotating-frame intermolecular double-quantum spin-lattice relaxation, T(1rho, DQC). A novel magnetic resonance imaging methodology based on intermolecular multiple-quantum coherences is demonstrated on a 7.05-T microimaging scanner. The results clearly reveal that the intermolecular double-quantum coherence T(1rho, DQC)-weighted imaging technique provides an alternative contrast mechanism to conventional imaging. PMID- 15799055 TI - Numerical solution of the Bloch equations provides insights into the optimum design of PARACEST agents for MRI. AB - Paramagnetic lanthanide complexes that display unusually slow water exchange between an inner sphere coordination site and bulk water may serve as a new class of MRI contrast agents with the use of chemical exchange saturation transfer (CEST) techniques. To aid in the design of paramagnetic CEST agents for reporting important biological indices in MRI measurements, we formulated a theoretical framework based on the modified Bloch equations that relates the chemical properties of a CEST agent (e.g., water exchange rates and bound water chemical shifts) and various NMR parameters (e.g., relaxation rates and applied B(1) field) to the measured CEST effect. Numerical solutions of this formulation for complex exchanging systems were readily obtained without algebraic manipulation or simplification. For paramagnetic CEST agents of the type used here, the CEST effect is relatively insensitive to the bound proton relaxation times, but requires a sufficiently large applied B(1) field to highly saturate the Ln(3+) bound water protons. This in turn requires paramagnetic complexes with large Ln(3+)-bound water chemical shifts to avoid direct excitation of the exchanging bulk water protons. Although increasing the exchange rate of the bound protons enhances the CEST effect, this also causes exchange broadening and increases the B(1) required for saturation. For a given B(1), there is an optimal exchange rate that results in a maximal CEST effect. This numerical approach, which was formulated for a three-pool case, was incorporated into a MATLAB nonlinear least square optimization routine, and the results were in excellent agreement with experimental Z-spectra obtained with an aqueous solution of a paramagnetic CEST agent containing two different types of bound protons (bound water and amide protons). PMID- 15799058 TI - Addressing efficiency and residual magnetization cross talk in multi-slice 2D steady-state free precession imaging of the heart. AB - This work presents an efficient method for achieving steady state in multi-slice 2D balanced steady-state free precession (SSFP) imaging of cardiac function. With current techniques, data acquisition for each slice is preceded by one or two heartbeats of dummy excitations. Depending on the number of heartbeats required for data acquisition, these dummy heartbeats can represent a large fraction of the total imaging time. As described here, FIESTA-SP (FIESTA with steady-state preparation) increases the imaging efficiency to nearly 100% by eliminating dummy heartbeats. Steady state for each slice is achieved using a linear flip angle series of excitations during the first cardiac phase of the first heartbeat for each slice. Because imaging proceeds immediately from one slice to the next, a heretofore-unseen issue arises where residual magnetization from each slice contaminates subsequent acquisitions. Accelerating the approach to steady state for each slice and eliminating slice cross talk are important for both multi slice and interactive real-time imaging. PMID- 15799059 TI - Development of a 0.014-inch magnetic resonance imaging guidewire. AB - The purpose of this study was to develop a standard 0.014-inch intravascular magnetic resonance imaging guidewire (MRIG), a coaxial cable with an extension of the inner conductor, specifically designed for use in the small vessels. After a theoretical analysis, the 0.014-inch MRIG was built by plating/cladding highly electrically conductive materials, silver or gold, over the inside and outside of the coaxial conductors. The conductors were made of superelastic, nonmagnetic, biocompatible materials, Nitinol or MP35N. Then, in comparison with a previously designed 0.032-inch MRIG, the performance of the new 0.014-inch MRIG in vitro and in vivo was successfully evaluated. This study represents the initial work to confirm the critical role of highly conductive and superelastic materials in building such small-size MRIGs, which are expected to generate high-resolution MR imaging of vessel walls/plaques and guide endovascular interventional procedures in the small vessels, such as the coronary arteries. PMID- 15799060 TI - Multislice and multicoil phase-sensitive inversion-recovery imaging. AB - Phase-sensitive inversion-recovery (PSIR) imaging may provide enhanced T(1) contrast. However, clinical implementation of PSIR imaging is hindered because image reconstruction with this method often lacks robustness and requires manual intervention, particularly for data acquired in multiple slices and with phased array coils. In this paper, a new algorithm suitable for automatic PSIR image reconstruction of multislice and multicoil data is presented. This algorithm phase corrects by region-growing, employing both the magnitude and the phase information of image pixels. Specifically, phase gradients of the original complex image are first calculated and then used to determine the sequence of the region-growing. The signal direction relating to the phase error for each pixel is then determined during the region-growing using both the magnitude and the phase of the previously determined pixels that are located within a boxcar neighborhood of the pixel. Finally, the intrinsic intercoil and interslice correlation is exploited to ensure consistency in the global polarity of all of the PSIR images. The results are demonstrated with in vivo human brain images acquired at 3 Tesla with an eight-channel phased-array coil. PMID- 15799061 TI - Mathematical framework for simulating diffusion tensor MR neural fiber bundles. AB - White matter (WM) fiber tractography (i.e., the reconstruction of the 3D architecture of WM fiber pathways) is known to be an important application of diffusion tensor magnetic resonance imaging (DT-MRI). For the quantitative evaluation of several fiber-tracking properties, such as accuracy, noise sensitivity, and robustness, synthetic ground-truth DT-MRI data are required. Moreover, an accurate simulated phantom is also required for optimization of the user-defined tractography parameters, and objective comparisons between fiber tracking algorithms. Therefore, in this study a mathematical framework for simulating DT-MRI data, based on the physical properties of WM fiber bundles, is presented. We obtained a model of a WM fiber bundle by parameterizing the various features that characterize this bundle. We then evaluated three different synthetic DT-MRI models using experimental data in order to test the proposed methodology, and to determine the optimum model and parameter settings for constructing a realistic simulated DT-MRI phantom. Several examples of how the mathematical framework can be applied to compare fiber-tracking algorithms are presented. PMID- 15799062 TI - Magnetic resonance imaging of inflammation with a specific selectin-targeted contrast agent. AB - E-selectin-targeted contrast enhancement of blood vessels in inflamed tissues was investigated with a new contrast agent, Gd-DTPA-B(sLe(x))A, which was recently obtained by grafting a synthetic mimetic of sialyl-Lewis(x), an E-selectin ligand, onto Gd-DTPA. The pharmacokinetics, biodistribution, and potential to image inflammation by MRI of this E-selectin-targeted contrast agent were evaluated. The inhibition (by 15-34%) produced by Gd-DTPA-B(sLe(x))A on Sialyl Le(x)-PAA-biotin binding to E-selectin confirmed the specific interaction of the new contrast agent with this adhesion molecule. Gd-DTPA-B(sLe(x))A was tested at a dose of 0.1 mmol/kg b.w. on mice and rats in a fulminant hepatitis model induced by the co-administration of D-galactosamine and E. coli lipopolysaccharide. A significant and prolonged contrast enhancement between blood vessels and liver parenchyma was obtained in pathological conditions, which attests to the specificity of the agent for E-selectin. The prolonged vascular residence (48.9 min in hepatitis vs. 29.8 min in healthy animals), as evidenced by the pharmacokinetic characterization, suggests that Gd-DTPA-B(sLe(x))A interacts with the specific receptors expressed during inflammation. The biodistribution of the compound indicates its retention in inflamed liver by both specific mechanisms and nonspecific accumulation due to the necrotic lesions. The same mechanisms are invoked to account for its retention in the spleen. PMID- 15799063 TI - Experimental measurement of extravascular parenchymal BOLD effects and tissue oxygen extraction fractions using multi-echo VASO fMRI at 1.5 and 3.0 T. AB - Quantitative interpretation of BOLD fMRI signal changes has predominantly employed empirical models for the whole parenchyma and a calibration step is usually needed to determine the physiological parameters during activation. Although analytical expressions are available for the extravascular and intravascular components of the BOLD effects, it is difficult to experimentally separate tissue from blood signal contributions at the low magnetic fields in which most fMRI studies are performed. Even if this can be achieved, an additional problem that remains is the separation of two types of extravascular BOLD effects, namely those around microvasculature (in the parenchyma close to the site of activation) and those around draining macrovasculature (e.g., in tissue and CSF more remote from the site of activation). In the recently developed vascular space occupancy technique, blood signals are nulled and the activations are localized predominantly in gray matter, allowing experimental measurement of parenchymal extravascular R(2)* and its changes accompanying activation. When comparing such data with total parenchymal R(2)* changes in BOLD fMRI, the extravascular fractions were found to be 47 +/- 7% (mean +/- SEM, n = 4) and 67 +/- 6% at 1.5 and 3.0 T, respectively, in line with expectations that intravascular BOLD contributions are reduced at higher field. The present approach provides a noninvasive means to determine parenchymal oxygen extraction fraction (OEF) in situ. During visual stimulation, OEF values measured at 1.5 and 3.0 T were in good agreement, giving 0.23 +/- 0.01 and 0.21 +/- 0.01, respectively. PMID- 15799064 TI - Multi-slice echo-planar spectroscopic MR imaging provides both global and local metabolite measures in multiple sclerosis. AB - MR spectroscopy (MRS) provides information about neuronal loss or dysfunction by measuring decreases in N-acetyl aspartate (NAA), a metabolite widely believed to be a marker of neuronal viability. In multiple sclerosis (MS), whole-brain NAA (WBNAA) has been suggested as a marker of disease progression and treatment efficacy in treatment trials, and the ability to measure NAA loss in specific brain regions early in the evolution of this disease may have prognostic value. Most spectroscopic studies to date have been limited to single voxels or nonlocalized measurements of WBNAA only, and longitudinal studies have often been hampered by standardization and reproducibility problems. Multi-slice echo-planar spectroscopic imaging (EPSI) is presented as a promising alternative to single voxel or nonlocalized spectroscopy for obtaining global metabolite estimates in MS. In the same session, measurements of metabolites in specific brain areas chosen after image acquisition (e.g., normal-appearing white matter (NAWM), gray matter (GM), and lesions) can be obtained. The identification and exclusion of regions that are inadequate for spectroscopic evaluation in global assessments can significantly improve quality and reproducibility, as demonstrated by a low within-subject variance in healthy controls. The reproducibility of the technique makes it a promising tool for future longitudinal spectroscopic studies of MS. PMID- 15799066 TI - Starter sequence for steady-state free precession imaging. AB - The dynamic equilibrium exploited by balanced steady-state free precession imaging develops slowly because its formation is dependent on both spin-spin and spin-lattice relaxation times. Attempting to image before steady state is established results in artifacts due to transient signal oscillations. Using a starter sequence to precondition the spin system can significantly reduce the delay before imaging. An improved design for a steady-state starter sequence is presented. The new sequence has the advantage of uniformly exciting the steady state response for all resonance offsets and can be phase cycled to suppress banding artifacts. PMID- 15799067 TI - Flip angle calculation for consistent contrast in spoiled gradient echo imaging. AB - In spoiled gradient echo sequences, the T(1)-weighting of image contrast is strongly affected by a nonlinear interaction of two sequence parameters, repetition time (TR) and flip angle (alpha). If alpha is not properly adjusted to compensate for variation in TR due to changing resolution, bandwidth, or number of slices, any optimization of contrast-to-noise may be compromised. Currently, there is no direct way to compare or reproduce the contrast properties of one sequence to another with a different TR. Here, it is demonstrated that for short TRs alpha may be calculated and automatically adjusted such that relative contrast--the shape of the signal-versus-T(1) curve--remains consistent and signal scales proportionally to radicalTR in all tissues. TR is then free to vary to accommodate a range of sequence parameters without impacting relative contrast and the T(1)-weighting of one sequence can be compared to, or reproduced in, another study with a different TR. PMID- 15799065 TI - Uncovering hidden in vivo resonances using editing based on localized TOCSY. AB - A novel single-shot spectral editing technique for in vivo proton NMR is proposed to recover resonances of low-concentration metabolites obscured by very strong resonances. With this new method, editing is performed by transferring transverse magnetization to J-coupled spins from selected coupling partners using a homonuclear Hartmann-Hahn polarization transfer with adiabatic pulses. The current implementation uses 1D-TOCSY with single-voxel localization based on LASER to recover the H1 proton of beta-glucose at 4.63 ppm from under water and the lactate methyl resonances from beneath a strong lipid signal. The method can be extended to further spin systems where conventional editing methods are difficult to perform. PMID- 15799068 TI - Exchange-influenced T2rho contrast in human brain images measured with adiabatic radio frequency pulses. AB - Transverse relaxation in the rotating frame (T(2rho)) is the dominant relaxation mechanism during an adiabatic Carr-Purcell (CP) spin-echo pulse sequence when no delays are used between pulses in the CP train. The exchange-induced and dipolar interaction contributions (T(2rho,ex) and T(2rho,dd)) depend on the modulation functions of the adiabatic pulses used. In this work adiabatic pulses having different modulation functions were utilized to generate T(2rho) contrast in images of the human occipital lobe at magnetic field of 4 T. T(2rho) time constants were measured using an adiabatic CP pulse sequence followed by an imaging readout. For these measurements, adiabatic full passage pulses of the hyperbolic secant HSn (n = 1 or 4) family having significantly different amplitude-and frequency-modulation functions were used with no time delays between pulses. A dynamic averaging (DA) mechanism (e.g., chemical exchange and diffusion in the locally different magnetic susceptibilities) alone was insufficient to fully describe differences in brain tissue water proton T(2rho) time constants. Measurements of the apparent relaxation time constants (T(2) (dagger)) of brain tissue water as a function of the time between centers of pulses (tau(cp)) at 4 and 7 T permitted separation of the DA contribution from that of dipolar relaxation. The methods presented assess T(2rho) relaxation influenced by DA in tissue and provide a means to generate T(2rho) contrast in MRI. PMID- 15799069 TI - Calculation of left ventricle relative pressure distribution in MRI using acceleration data. AB - Measurements of pressure variations within the cardiac chambers could provide important information for clinical assessments of cardiovascular function. In this work an MRI method for evaluating spatial distributions of intracardiac relative pressure is presented. We first calculated pressure gradients from MR maps of blood acceleration by applying the NS equation. We then used an original algorithm to compute pressure distribution in a region of interest (ROI) by minimizing the pressure gradient curl so that the result in a given pixel is independent of the integration path. The method was assessed in five healthy volunteers by means of MR 2D maps of the blood acceleration in the left ventricle (LV) during ejection and filling phases. The pressure variations calculated from acceleration mapping fit the known physiological variations better than those based on velocity maps acquired in the same volunteers. Furthermore, the optimization algorithm presented here produced the same results as iterative algorithms proposed by other authors, but in much less time and without requiring adjustable parameters or boundary conditions. PMID- 15799070 TI - Identification of degradation products formed during performic oxidation of peptides and proteins by high-performance liquid chromatography with matrix assisted laser desorption/ionization and tandem mass spectrometry. AB - Oxidation of proteins with performic acid is extensively used to cleave disulfide bonds. Due to its efficiency and many other advantages it deserves more attention especially in proteomics as a method for sample treatment. However, some unwanted degradations can occur during performic oxidation. In this work the degradation products during performic oxidation of two peptides and bovine serum albumin as model substrates were explored by coupling high-performance liquid chromatography (HPLC) to matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI-TOF/TOFMS). In addition to well-known modifications such as oxidation of tryptophan and oxidation and chlorination of tyrosine, novel degradation products including nonspecific cleavage after asparagine or tryptophan, formylation of lysine, and beta-elimination of cysteine, were observed. Although almost all of these modification/degradation products except oxidation products of tryptophan were formed at sub-stoichiometric levels, they can cause confusion as a result of the sensitivity of mass spectrometry in analysis of the oxidized samples, especially in proteomics research. The results presented here will facilitate the interpretation of analytical data for performate-oxidized samples, and help to select appropriate methods for each unique sample. PMID- 15799071 TI - Determination of urinary S-phenylmercapturic acid, a specific metabolite of benzene, by liquid chromatography/single quadrupole mass spectrometry. AB - A high-performance liquid chromatography/single quadrupole mass spectrometry (LC/MS) method is described for the determination of urinary S-phenylmercapturic acid (S-PMA), a specific metabolite of benzene. Urine samples were spiked with [13C6]S-PMA (used as the internal standard) and acidified; then they were purified by solid-phase extraction (SPE) on C18 cartridges. Analyses were conducted on a reversed-phase column by gradient runs with 1% aqueous acetic acid/methanol mixtures at different proportions as the mobile phase. The detector was used in electrospray negative ion mode (ESI-), the ions m/z 238 for S-PMA and 244 for [13C6]S-PMA being recorded simultaneously. The detection limit (for a signal-to-noise ratio = 3) was 0.2 microg/L, thus allowing for the measurement of background excretion of S-PMA in the general population. The use of the internal standard allowed us to obtain good precision (CV% values < 3%) and a linear calibration curve within the range of interest for monitoring occupational exposure to benzene (up to 500 microg/L). The method was applied to assay the metabolite concentration in a group of 299 workers (68 smokers and 231 non smokers) occupationally exposed to relatively low levels of benzene (environmental concentration = 0.4-220 microg/m3, mean 11.4 microg/m3 and 236 non exposed subjects (134 smokers and 102 non-smokers). The results clearly showed that smoking must be taken into account for the correct interpretation of the results of S-PMA measurements for the assessment of work-related benzene exposure. When only non-smokers were selected, the mean excretion of S-PMA was significantly higher in workers exposed to benzene (1.2 +/- 0.9 microg/g creatinine) than in the control group (0.7 +/- 0.6 microg/g creatinine) (p < 0.001), thus confirming the role of S-PMA as a biomarker of benzene on a group basis, even for relatively low exposure degrees. PMID- 15799072 TI - Screening for dementia in an Irish community sample using MMSE: a comparison of norm-adjusted versus fixed cut-points. AB - BACKGROUND: The MMSE is a widely-used instrument in screening for dementia in the community. The traditional cut-point of <24/30 may be unsuitable for use in some settings, due to biases in age and education. OBJECTIVES: This study aimed to investigate whether cut-points derived from age- and education-specific norms would improve the performance of the MMSE as a screening tool for dementia and cognitive impairment in an Irish community setting, potentially reducing the number of onward referrals. A secondary aim was to obtain score norms in an Irish sample. METHODS: One thousand one hundred and fifteen people aged over 65 (mean age 74.8+/-6.8 years; 68% female) were assessed in their own homes, using MMSE (world) and the AGECAT computerised diagnostic system for mental illness in the elderly. The performance of the MMSE in identifying case- and sub-case-level dementia was investigated using cut-points of <24 and <23, as well as adjusted cut-points based on published norms and norms derived from the Irish sample. RESULTS: Published norms did not prove useful in improving screening accuracy. Cut-points based on 10th percentile Irish norms and the <23 cut-point performed comparably well, both yielding 93% specificity, with overall accuracy of 90% and 91% respectively (as sensitivity was higher for the <23 cut-point). CONCLUSIONS: Locally-derived norms yielded better screening accuracy than did published norms or the traditional <24 cut-point. The importance of selecting an appropriate percentile cut-off when using norms is discussed. The best results were obtained with a simple <23 cut-point, and this may be optimal when screening for dementia in an Irish community setting. PMID- 15799073 TI - Enantiomer ratio of MK-0767 in humans and nonclinical species. AB - MK-0767, (+/-)-5-[(2,4-dioxothiazolidin-5-yl)methyl]-2-methoxy-N-[[(4 trifluoromethyl)phenyl]methyl]benzamide, is a thiazolidinedione-containing dual peroxisome proliferator-activated receptor (PPAR) alpha/gamma agonist that has been studied as a potential treatment for patients with type 2 diabetes. MK-0767 contains a chiral center at the C-5 position of the thiazolidinedione ring and was being developed as the racemate, due to the rapid interconversion of its enantiomers in biological samples. In the present work the in vitro and in vivo concentration ratios of the (+)-(R) to (-)-(S) enantiomers of MK-0767 were determined in plasma from humans (in vitro only) and nonclinical species used in the toxicological evaluation of rac-MK-0767, namely CD-1 mice, Sprague-Dawley rats, beagle dogs, New Zealand white rabbits, and rhesus monkeys. The R/S ratio was determined by chiral liquid chromatography/tandem mass spectrometry. Species differences were observed in the in vitro and in vivo enantiomeric ratios, as well as differences between in vitro and in vivo in some species. The in vitro R/S ratio was similar in dogs and humans (approximately 1.5-1.7). In rats and monkeys, the ratio was approximately unity, both in vitro and in vivo. In mice, the ratio was higher in vitro (approximately 1) than in vivo (approximately 0.6), while in rabbits it was higher in vivo (approximately 1) than in vitro (approximately 0.5). These results suggested that differential binding of the MK 0767 enantiomers to plasma and tissue proteins and other macromolecules may be affecting the R/S ratio both in vitro and in vivo, since in protein-free systems MK-0767 exists as the racemate. PMID- 15799074 TI - The fast track day hospital: improved care of patients with dementia and associated behavioural disorder. PMID- 15799075 TI - Survival of a cohort of elderly patients with advanced dementia: nasogastric tube feeding as a risk factor for mortality. AB - OBJECTIVES: To evaluate and identify factors determining survival in elderly patients with advanced dementia. METHODS: A prospective, follow-up, observational analysis in a cohort of 67 community-based patients aged 65 years or older with dementia defined by DSM-IV and stage 7A or above on the FAST scale. Data were recorded on socio-demographic variables, FAST, Katz index, language, swallowing ability, diet, nutritional status (from anthropometric and laboratory data), associated diseases and medical complications during the previous 12 months. Survival was analyzed by the Kaplan-Meier method. Prognostic factors for survival were identified by the Cox proportional hazards regression model. RESULTS: The median follow-up was 832 days. The mean age was 82.2+/-6.7 years and 92.5% were women. A comorbid condition was present in 71.6%, most frequently hypertension (22.4%). A clinical event had occurred in 52 (77.6%) patients during the previous year (pneumonia, urinary infection, stroke, pressure sore, dehydration, sepsis or others). A total of 25 (37.3%) patients died. The mean survival was 676 days (95% confidence interval, 600-752 days). Cox proportional hazards model showed that independent prognostic factors for mortality were having pneumonia within the previous year (RR:3.7; p=0.001), a permanent nasogastric tube (RR:3.5; p=0.003) and serum albumin values below 3.5 g/dL (RR:2.9; p=0.028). CONCLUSIONS: In patients with advanced dementia, hypoalbuminemia and pneumonia are strongly and positively associated with mortality. Artificial nutrition via a nasogastric tube reduces survival in these patients. PMID- 15799077 TI - Utility of a Scottish regional intensive psychiatric care unit for demented elders. PMID- 15799076 TI - Prevalence and correlates of depression in late life: a population based study from a rural Greek town. AB - BACKGROUND: Depression in late life is common and has serious consequences on function, medical co-morbidity, quality of life, and use of medical services. OBJECTIVE: To estimate the age- and gender-specific prevalence of depression among people over 60 years of age, and to examine correlates of depression, in particular the relationship between depression and cognitive impairment. METHOD: From a total of 965 inhabitants, aged over 60 years, in Velestino, a rural town in central Greece, 608 were accessible and constituted the target population. During a five-month period in 2000, a trained health visitor interviewed all study participants. The interview covered socio-demographic characteristics, medical history, and administration of the 15-question Geriatric Depression Scale (GDS-15) and the Mini Mental Scale Examination instrument (MMSE). RESULTS: The prevalence of mild or more severe depression (GDS> or =7) was 27%, while the prevalence of moderate to severe depression (GDS> or =11) was 12%. Increasing age, female gender, lower education, and being currently unmarried were associated with higher risk of depression in univariate regression models, but these associations disappeared after controlling for cognitive function, except for the association with marital status. Cognitive impairment was strongly associated with increased risk for depression. The co-morbid presence of digestive, neurological and heart conditions was also associated with increased risk for depression, while cancer was not. CONCLUSION: In a rural Greek area, the prevalence of depression in late life is high. Depression was more common among unmarried individuals, those with significant cognitive impairment, and in association with specific medical conditions. PMID- 15799078 TI - Needs assessment in dementia. AB - BACKGROUND: Resource allocation and service development traditionally focuses on diagnostic categories and consequent perceived need. Identification of the actual level of need in the elderly with dementia, and the degree to which it is unmet is necessary to plan services both individually and as a group. The aim of this study was to characterise the needs of a sample of community dwelling elderly patients with dementia who were referred to an old age psychiatry service in Ireland between July 2002 and July 2003. METHODOLOGY: Eighty-two consecutively referred community dwelling patients with ICD-10 diagnosis of dementia were assessed on The Care Needs Assessment Pack for Dementia (CareNap-D). Data on needs across seven domains (health and mobility, self-care and toileting, social interaction, thinking and memory, behaviour and mental state, housecare, community living) is presented (Reynolds T et al., 1998). RESULTS: Subjects had a mean of 33 (range: 13-56) identified needs. Approximately 1/3 of these were unmet with a mean of 13 (range: 0-37) and a mean of 20 (range: 4-39) were met. High levels of unmet need was identified in the domains of behaviour and mental state (84% of those with agitation) and of social interaction (79% of those with 'partaking in activities' need). The specific item of repetitive questioning occurred in 68 individuals and was unmet in 88% of these cases. Increasing age, lower MMSE score, and living alone were associated with greater total levels of unmet need. CONCLUSION: This data underlines the degree of unmet need in the community dwelling elderly with dementia and the importance of developing a spectrum of services on the basis of the actual needs identified. PMID- 15799079 TI - The effect of integrated emotion-oriented care versus usual care on elderly persons with dementia in the nursing home and on nursing assistants: a randomized clinical trial. AB - OBJECTIVES: To examine the effect of integrated emotion-oriented care on nursing home residents with dementia and nursing assistants. DESIGN: A multi-site randomized clinical trial with matched groups, and measurements at baseline and after seven months. SETTING: Sixteen psychogeriatric wards in fourteen nursing homes in the Netherlands. PARTICIPANTS: One hundred and forty-six elderly residents with the diagnosis dementia of the Alzheimer (DAT) type, mixed DAT and vascular dementia, and dementia syndrome (NAO) and 99 nursing assistants. INTERVENTIONS: Integrated emotion-oriented care and usual care. MESUREMENTS: Demented elderly: Behaviour and mood related to adaptation to the illness and the institutionalization. Nursing assistants: General health as measured by feelings of stress, stress reactions, feeling of competence and illness. RESULTS: Positive effects in favour of the integrated emotion-oriented care were found in mild to moderately demented residents on two adaptive tasks: maintaining an emotional balance (less anxiety) and preserving a positive self-image (less dissatisfaction). In the trained group of nursing assistants fewer stress reactions were found only in those who perceived improvement in their emotion oriented care skills after training. CONCLUSION: Emotion-oriented care is more effective with regard to the emotional adaptation in nursing homes of persons with a mild to moderate dementia. For the severely demented elderly we did not find this surplus value. This outcome is of clinical importance for elderly persons with dementia who are cared for in nursing homes. With respect to the nursing assistants it is concluded that emotion-oriented care has a positive influence on stress reactions in some of them. PMID- 15799080 TI - Factors affecting timely recognition and diagnosis of dementia across Europe: from awareness to stigma. AB - BACKGROUND: Timely recognition and diagnosis of dementia is the pre-condition for improving dementia care, but diagnosis often occurs late in the disease process. OBJECTIVE: To compare facilitators and obstacles to the timely recognition of dementia across eight European Union states, in order to implement established policies for earlier diagnosis. METHODS: A modified focus group technique, including a pre and posterior procedure. RESULTS: Twenty-three participants from different disciplines, purposively sampled for professional expertise in dementia research and innovative practice, attended two focus groups. Stigma in ageing and dementia, accompanied by a sense that there is little to offer until later on in the disease, underpinned the widespread reluctance of GPs to recognise dementia at an early stage and were major obstacles to the timely diagnosis of dementia across all eight countries. Dementia care services varied widely across Europe. Countries with the greatest development of dementia health care services were characterised by national guidelines, GPs fulfilling a gatekeeper function, multi disciplinary memory clinics and innovative programmes that stimulated practice and new services. Dementia-related stigma was perceived as being less prominent in these countries. CONCLUSIONS: Overcome of delays in the timely diagnosis of dementia needs more than specialist services. They should address the processes associated with stigma, age and dementia, especially where these relate to physician practice and diagnostic disclosure. Stigma is perceived as variable across European States, with a promising finding that its impact is relatively small in countries with the widest range of dementia care services. PMID- 15799081 TI - The effects of psychosocial methods on depressed, aggressive and apathetic behaviors of people with dementia: a systematic review. AB - OBJECTIVES: This systematic review seeks to establish the extent of scientific evidence for the effectiveness of 13 psychosocial methods for reducing depressed, aggressive or apathetic behaviors in people with dementia. METHODS: The guidelines of the Cochrane Collaboration were followed. Using a predefined protocol, ten electronic databases were searched, studies selected, relevant data extracted and the methodological quality of the studies assessed. With a Best Evidence Synthesis the results of the included studies were synthesized and conclusions about the level of evidence for the effectiveness of each psychosocial method were drawn. RESULTS: There is some evidence that Multi Sensory Stimulation/Snoezelen in a Multi Sensory Room reduces apathy in people in the latter phases of dementia. Furthermore, there is scientific evidence, although limited, that Behavior Therapy-Pleasant Events and Behavior Therapy Problem Solving reduce depression in people with probable Alzheimer's disease who are living at home with their primary caregiver. There is also limited evidence that Psychomotor Therapy Groups reduce aggression in a specific group of nursing home residents diagnosed with probable Alzheimer's disease. For the other ten psychosocial methods there are no or insufficient indications that they reduce depressive, aggressive or apathetic behaviors in people with dementia. CONCLUSIONS: Although the evidence for the effectiveness of some psychosocial methods is stronger than for others, overall the evidence remains quite modest and further research needs to be carried out. PMID- 15799082 TI - The factor structure of the Hospital Anxiety and Depression Scale in older individuals with acquired amputations: a comparison of four models using confirmatory factor analysis. AB - BACKGROUND: There has been little attention to the underlying dimensional structure of the Hospital Anxiety and Depression Scale (HADS) in analyses involving individuals older than 65 years of age despite its routine application in this age group. METHODS: The factor structure of the HADS was investigated using a sample of 680 veterans with limb amputations who were aged at least 66 years (mean 79.0, SD 5.02; range 66-92) Four models were specified and estimated using Lisrel 8.54. Model 1 specified a uni-dimensional structure. Model 2 specified a correlated two-factor model. Model 3 specified a correlated two factor model but with cross factor loadings for item 7. Model 4 specified a three factor model after Clark and Watson's (1991) tripartite theory of anxiety and depression. RESULTS: Models 2, 3 and 4 were found to adequately fit the data. However, on balance, model 4 was found to be a better explanation of the data than the alternative specifications. CONCLUSIONS: The results indicate that factor models for the HADS developed in younger samples are replicable with older adults, thus supporting the suggestion that the HADS structure is invariant for age. However, considering the composition of the current sample, i.e. veterans with limb amputations, further research is necessary to determine whether these findings are generalisable to the wider population of older adults. PMID- 15799083 TI - The sensitivity of somatic symptoms in post-stroke depression: a discriminant analytic approach. AB - BACKGROUND: Somatic and neurocognitive symptoms of depression may overlap with the physical symptoms of stroke, and thus make the diagnosis of post-stroke depression difficult. AIM: To assess the sensitivity of individual depressive symptoms and their contribution to the diagnosis of post-stroke depression. PATIENTS AND METHODS: Two hundred and six patients with first-ever stroke, participating in a longitudinal study, were administered the Structured Clinical Interview for DSM-IV and the Hamilton Depression Rating Scale (HAM-D). In a discriminant analysis the relative contribution of the individual HAM-D items to the diagnosis of major depressive disorder was evaluated. RESULTS: The cumulative incidence of post-stroke major depressive disorder was 32%. The discriminant model based on HAM-D item scores was highly significant (p<0.001) and classified 88.3% of patients correctly as depressed or nondepressed. As expected, 'depressed mood' discriminated best between depressed and non-depressed stroke patients. 'Reduced interests' had a relatively low sensitivity and may in part reflect 'apathy', which often is considered a separate construct. With the exception of 'suicidal thoughts', most psychological symptoms, such as 'hypochondriasis', 'lack of insight' and 'feelings of guilt', were not very sensitive. Some somatic symptoms, such as 'reduced appetite', 'psychomotor retardation', and 'fatigue' had high discriminative properties. CONCLUSION: Psychological, neurocognitive and somatic symptoms of depression differ among themselves in terms of diagnostic sensitivity, and should be considered individually. Some somatic symptoms are highly sensitive for depression and should not be neglected by following an 'exclusive' or 'attributional' approach to the diagnosis of PSD. PMID- 15799084 TI - Group III metabotropic glutamate receptor activation inhibits Ca2+ influx and nitric oxide synthase activity in bone marrow stromal cells. AB - Nitric oxide (NO) is pivotal to bone physiology. In the central nervous system constitutive, Ca(2+)-calmodulin regulated NO synthase activity and glutamate signalling are intimately linked. Since L-glutamate signalling occurs in bone and is implicated in bone regulation, we have investigated the effect of L-glutamate on NO synthase in bone-derived cells. Treatment of marrow stromal cells with L glutamate reduced basal NO synthase activity by 40%. Imaging showed that L glutamate caused a rapid, usually localised and slowly-reversible fall in [Ca(2+)](i). This effect was resistant to disruption of intracellular Ca(2+) stores but sensitive to extracellular La(3+) or omission of extracellular Ca(2+), demonstrating that glutamate acts by inhibition of membrane Ca(2+) influx. The only previous description of such an effect of L-glutamate is via activation of the group III receptor, mGluR6, in the retina. Using Western blotting and RT-PCR we detected mGluR6 protein and transcripts in marrow stromal cells. The effects of L-glutamate on NOS activity and [Ca(2+)](i) in marrow stromal cells were abolished by a group III mGluR inhibitor, (S)-2-amino-2-methyl-4-phosphonobutyric acid. Recording of membrane potential showed that, similarly to the effects of retinal mGluR6 activation, L-glutamate induced membrane hyperpolarisation (-16 +/ 2 mV), which was also sensitive to group III mGluR inhibition. L-glutamate had no effect on cAMP levels. We conclude that activation of a group III mGluR in bone marrow stromal cells inhibits a Ca(2+)-permeable plasma membrane channel, reducing [Ca(2+)](i) and suppressing generation of NO. These observations directly link bone L-glutamate signalling to processes central to bone growth and regulation. PMID- 15799085 TI - Comparison of the prevalence and risk factors for depressive symptoms among elderly nursing home residents in Taiwan and Hong Kong. AB - BACKGROUND: Depression in the elderly has become a serious health care issue worldwide. However, no studies have compared the prevalence and risk factors for depressive symptoms among institutionalized Chinese elders living in different regions. OBJECTIVES: To explore and compare the prevalence and risk factors for depressive symptoms among elderly residents of nursing homes in Taiwan and Hong Kong. METHODS: Random sampling was used to recruit participants from eight nursing homes in each region. A total of 150 elders from Taiwan and 214 elders from Hong Kong participated. RESULTS: The prevalence of depressive symptoms was significantly higher in participants from Hong Kong (65.4%) than in Taiwan (43.3%). Logistic regression analysis indicated that gender, satisfaction with living situation, perceived health condition, and perceived income adequacy significantly predicted depressive symptoms in elderly nursing home residents in Taiwan. Significant predictors of depressive symptoms in the Hong Kong sample were satisfaction with living situation, cognitive status, and functional status. CONCLUSIONS: It is important to consider risk factors specific to a target population when developing depression intervention programs. PMID- 15799087 TI - When the "last gasp" is an SOS. PMID- 15799086 TI - Going downhill can be healthy for your heart. PMID- 15799088 TI - Ask the doctors. I've read that the medication spironolactone may cause cardiac arrest. Please discuss the pros and cons of this medicine. Does The Cleveland Clinic prescribe such a regimen for conditions after heart surgery? PMID- 15799090 TI - Massage and health. Hands-on care. PMID- 15799089 TI - Ask the doctors. I had a heart attack two years ago. Since then, I have been taking a statin, and my cholesterol numbers are excellent. My chiropractor suggests I should be taking CoQ10 along with my statin for even better cholesterol health. Does that make sense? PMID- 15799091 TI - Moderate weight loss. Many health benefits. PMID- 15799092 TI - I know that some ways of preparing fish add fat and calories. Are there other health considerations when choosing how to prepare fish? PMID- 15799094 TI - Protecting the mental health of Gaza's inhabitants. PMID- 15799093 TI - The trials of tenofovir trials. PMID- 15799095 TI - Fighting to close the condom gap in Uganda. PMID- 15799096 TI - Are cardiac surgeons' mortality rates helpful? PMID- 15799097 TI - Clinical research is essential to help achieve Europe's goals. PMID- 15799098 TI - Throwing light on nipple discharge. AB - Five percent of the patients presenting to a symptomatic breast clinic have nipple discharge. Conventional surgical management for the nipple discharge includes microdochectomy or total duct excision. Breast duct micro-endoscopy (BDME) is a new technique, which helps evaluate the underlying cause of nipple discharge. We describe a case of nipple discharge with a unique etiology: mammary duct foreign body. PMID- 15799099 TI - The evolving role of DNA demethylating agents in the treatment of sickle cell disease and thalassemia. PMID- 15799100 TI - Asthma inhalers may pose risks in the milk-allergic child. PMID- 15799101 TI - Hepatitis B--another vaccine success story. PMID- 15799102 TI - Navigating the world of allergy and asthma products. PMID- 15799103 TI - Product recalls. Recalls: girls' hooded sweaters with drawstrings...and Fargglad multi-color children's chair. PMID- 15799104 TI - Breakfast and your health. PMID- 15799105 TI - Prostate cancer surgery: is nerve-sparing safe? PMID- 15799106 TI - Stress and the heart: treatment helps. PMID- 15799107 TI - On call. I am a 71-year-old man with diabetes and high blood pressure. I take Glucophage for my diabetes and Zestril for my blood pressure, and my doctor says both problems are under good control, but I've developed a very annoying problem. Every time I eat, I develop terrible sweating on my face and neck. It only lasts 10-15 minutes, but my shirt collar gets soaked and I'm too embarrassed to go out to dinner. Am I allergic to something in food? Am I getting hot flashes as my wife did when she had the change of life? PMID- 15799108 TI - Falling apart: dissociation and it disorders. How can psychotherapy heal divided minds? PMID- 15799109 TI - Treating opiate addiction, Part II: alternatives to maintenance. PMID- 15799110 TI - The treatment of attention deficit disorder: new evidence. PMID- 15799111 TI - Long-distance psychotherapy. PMID- 15799112 TI - What is modafinil? PMID- 15799113 TI - Nanometric position and displacement measurement of the six degrees of freedom by means of a patterned surface element. AB - A method is presented for position and displacement measurements of the six degrees of freedom by use of a patterned surface element observed by a static interferometric vision system. The surface element is made of a regular pattern of circular holes etched on a chromium layer deposited onto a flat glass plate. The in-plane coordinates (x, y, theta(z)) are reconstructed with a subpixel resolution by a vision method based on phase measurements. The out-of-plane coordinates (z, theta(x), theta(y)) are reconstructed by phase-shifting interferometry. Resolutions achieved by the proposed method are in the range of microradians for the measurement of angles and of nanometers for the position and displacement. PMID- 15799114 TI - Numerous budget proposals would affect group practice. PMID- 15799115 TI - Of death, drugs, recalls, and arrogance. PMID- 15799116 TI - Functional nervous disturbances. PMID- 15799117 TI - Professional practice solutions.... How does a healthcare organization deal with conflicting directives between HCFA regulations and Joint Commission standards when obligated to abide by both? PMID- 15799118 TI - Colon and rectal surgery. PMID- 15799119 TI - Medical toxicology. PMID- 15799120 TI - [Endocarditis on pulmonary valve by Enterococcus faecalis]. PMID- 15799121 TI - An item response analysis of the international restless legs syndrome study group rating scale for restless legs syndrome. AB - BACKGROUND AND PURPOSE: Restless legs syndrome (RLS) is a common central nervous system disorder; however, there is currently a lack of well-validated and easily administered measures of RLS severity available. The International Restless Legs Syndrome Study Group has recently developed a 10-item scale to meet this need. The International Restless Legs Severity Scale (IRLS) has been shown to have a high degree of reliability, validity, and internal consistency. In order to further demonstrate the validity of the IRLS, the present study examined the relationship between scores on individual IRLS items and overall RLS severity. PATIENTS AND METHODS: The 10-item IRLS was administered to 196 RLS patients. Option characteristic curves (the probability of scoring different options for a given item as a function of overall IRLS score) were generated in order to illustrate the scoring patterns for each item across the range of total RLS severity. Item characteristic curves (the expected score on an item as a function of overall IRLS score) were also generated to illustrate the relationship between scores on the individual items and total RLS severity. RESULTS: The IRLS items demonstrated excellent item response properties, with option and item characteristic curves closely approximating those of an ideal item. Item 3 (relief of arm or leg discomfort from moving around) was the most problematic item in that a 'floor' effect was evident; however, the item response characteristics for this item were still acceptable. CONCLUSIONS: Each IRLS item showed a good relationship between responses on that item and overall RLS severity, providing further evidence for the validity of the IRLS as a measure of RLS severity in RLS patients. PMID- 15799122 TI - The influence of sociodemographic factors on awareness, knowledge and attitude toward andropause among health professionals in Ile-Ife, Nigeria. AB - A descriptive study of the awareness, knowledge and attitude of health professionals toward andropause was conducted in Ile-Ife, Nigeria with the aim of assessing the influence of sociodemographic variables of the respondents on their perspectives of the subject matter. The study employed a structured questionnaire to assess respondents' level of awareness and knowledge, and Likert-type scales to rate respondents' attitudes. A total of 187 (45%) respondents indicated previous awareness of andropause, with younger people (aged below 40 years) displaying better awareness compared with the older ones (p = 0.05), and more doctors than 'other technical health professionals' displaying better awareness when compared with health administrators (p < 0.001). However, only 93 (23%) respondents demonstrated a good knowledge of andropause, with more females compared with males recording good knowledge scores (p = 0.01). While a slightly higher proportion of older respondents (aged 40 years and above) compared with younger ones demonstrated good knowledge of andropause, age and marital status were not significantly related to knowledge of the subject matter. While only 23 (5.4%) respondents displayed a positive attitude toward andropause, and respondents' knowledge was found to positively influence their attitude toward it, none of the sociodemographic variables of age, sex or marital status was significantly related to respondents' attitudes. The study concluded that there is still a low level of awareness and knowledge of andropause among health workers in Nigeria, unlike what obtains in more developed countries of the world, and called for active education of both health professionals and the general public on the subject matter of andropause and other related male reproductive health concerns in the country. PMID- 15799123 TI - Reflections concerning biochemical parameters of androgenicity. AB - Due to the highly variable concentration of sex hormone binding globulin (SHBG) and the many factors affecting it, the evaluation of the androgen status may require the measurement of a parameter of bioactive plasma testosterone. As, however, no practical, clinical useful direct method for measurement of plasma androgen bioactivity is available, indirect biochemical parameters are used. All have their limitations and pitfalls. In this paper are discussed some of the factors influencing the values obtained with different methods (direct measurement of free testosterone by analog radioimmunoassay, dialysis, ammonium sulfate precipitation, free testosterone index, calculated free and bioavailable testosterone), all of which may explain the variability of data reported in the literature. It is concluded that, whereas determination of bioavailable testosterone by dialysis or ammonium sulfate precipitation of SHBG-bound testosterone is work-intensive and not really suitable for clinical routine, while direct measurement of free testosterone by analog immunoassay yields unreliable results, only the free testosterone index and calculated bioavailable testosterone are adapted for clinical routine. The limitations of the free testosterone index, a dimensionless parameter which does not reflect a defined bioavailable testosterone concentration, are discussed. As the same measurements of testosterone and SHBG required for determination of the free testosterone index permit the calculation of bioavailable testosterone, which yields a defined androgen concentration, it is advisable to prefer the latter over the free testosterone index, which should no longer be used. PMID- 15799124 TI - Clinical effects of growth hormone on bone: a review. AB - Growth hormone (GH) stimulates bone turnover. Deficiency of GH due to hypopituitarism is related to low bone mineral density and increased fracture risk. GH substitution increases and thus normalizes bone mineral density in these patients, which is one of a number of arguments for GH substitution in hypopituitarism. In contrast, a possible therapeutic use of GH in idiopathic osteoporosis and glucocorticoid-induced osteoporosis is speculative and not established. Reduction of osteoporosis risk is an argument brought up for a use of GH in healthy elderly persons (anti-aging medicine). However, since only very limited data are available yet, this cannot be based on scientific evidence, and there are important concerns about the safety of use of GH in healthy elderly persons. PMID- 15799125 TI - Andropause (or symptomatic late-onset hypogonadism): facts, fiction and controversies. AB - The existence of the so-called 'andropause' is an irrefutable fact, although the terms 'SLOH' (symptomatic late-onset hypogonadism) or 'symptomatic ADAM' (androgen deficiency of the aging male) are more accurate. The term 'andropause' is, in most cases, inappropriate, except when the gonads cease functioning. Testosterone production decreases as a function of age, but this decrease is not universal. Several clinical manifestations are associated with hypogonadism, but these are not solely attributable to hypogonadism. Other hormones (i.e. dehydroepiandrosterone, growth hormone, thyroxine and melatonin) also decrease with age. Such multi-hormone alterations are closely inter-related and may influence 'andropause-related' symptoms. Many patients with SLOH, although by no means all, respond well to testosterone therapy. Although testosterone therapy can induce adverse effects, these can be largely minimized by proper monitoring by a knowledgeable clinician. Extrapolation of the effects of estrogens + progesterone in menopausal women to the use of testosterone in hypogonadal men is mythical, and more research on the effect of exogenous sex steroids in aging men and women is needed. However, to restrict the prescription of such hormones until all issues have been fully addressed is impossible. Indeed, the discourse on SLOH would benefit considerably from more science and less speculation. PMID- 15799126 TI - Clinical experiences with testosterone therapy: prostate safety. AB - Due to a decrease in Leydig cell function, a considerable proportion of men over 50 years of age will develop hypogonadism. Consequently, loss of libido and several other testosterone-dependent symptoms may become evident. When decreased levels of biologically available testosterone are found, and corresponding symptoms are present, these men could be eligible for testosterone substitution therapy. Testosterone treatment in testosterone-deprived men has been shown to improve general well-being, osteoporosis, muscle atrophy, libido and--if present- anemia. Despite these positive effects, testosterone treatment has to be performed with caution. Although it has not been proven that elevation of the serum testosterone level to the normal range results in a greater risk of developing prostate cancer, the effects of testosterone on a prostate cancer already present are well established. Several studies have demonstrated that testosterone treatment does not result in a significant increase in serum levels of prostate-specific antigen (PSA) or prostate volume. The long-term effects, however, are currently unknown. For these reasons, testosterone treatment should be performed only when the presence of prostate cancer is unlikely; i.e. when PSA levels are within normal limits and digital rectal examination does not reveal any suspicious findings. These examinations may still miss some small prostate cancers that could be promoted by testosterone treatment. The determination of PSA levels under testosterone treatment is necessary every 3 months, at least for the first year. Steadily rising PSA levels require immediate cessation of testosterone administration and the initiation of further diagnostic procedures (prostate biopsy), to rule out prostate cancer. PMID- 15799127 TI - Testosterone therapy in erectile dysfunction. AB - Studies in animals have indicated that the nitric oxide erectile pathway is testosterone-dependent. Castration induces erectile dysfunction and a reduction in nitric oxide synthase-stained nerves in erectile tissue. Furthermore, castration adversely affects penile hemodynamics and smooth muscle content, leading to veno-occlusive dysfunction. Testosterone replenishment reverses these physiological, biochemical and structural changes. Several clinical studies have demonstrated the benefits of a combination of testosterone and sildenafil. A recently published, multicenter study evaluated the safety and efficacy of testosterone gel 1% (Testogel; Schering AG, Germany/AndroGel; Solvay Pharmaceuticals) vs. placebo gel in conjunction with sildenafil, in producing an erectile response in hypogonadal men who did not respond to treatment with sildenafil alone for erectile dysfunction. The selection criteria required subjects to have had erectile dysfunction for at least 3 months, to be non responsive to 100 mg sildenafil and to have low testosterone levels (< 400 ng/dl). The primary efficacy measurement was the mean change from baseline in the Erectile Function domain of the International Index of Erectile Function (IIEF). Secondary outcome measures included the mean change from baseline in the other domains and the total sum of the IIEF. Subjects were randomized to receive either testosterone gel + sildenafil, or placebo gel + sildenafil for 12 weeks. Testosterone therapy with testosterone gel improved the erectile response to sildenafil. Therefore, testosterone therapy may be considered for the treatment of erectile dysfunction in men with low to low-normal testosterone levels, who have failed prior treatment with sildenafil alone. Consequently, it is important to screen for hypogonadism in men who fail PDE5 inhibitors. PMID- 15799128 TI - Testosterone therapy--what, when and to whom? AB - Testosterone therapy has been used for more than 60 years in the treatment of male hypogonadism. The classical forms of hypogonadism are comprised of primary testicular failure or insufficient testicular stimulation due to the lack of pituitary gonadotropins. Typical causes of primary hypogonadism are Klinefelter's syndrome, anorchia or acquired disturbances of testicular function. Secondary hypogonadism is characterized by insufficient production of pituitary gonadotropins, due either to pituitary failure or defects at the hypothalamic level. It is unequivocally accepted in clinical practice that any male with inadequately low testosterone production for his age will require androgen therapy. In addition to the classical forms of hypogonadism, the past decade of research has clearly demonstrated that, with increasing age, many men will suffer from decreasing testosterone production. About 15-25% of men over the age of 50 years will experience serum testosterone levels well below the threshold considered normal for men between 20 and 40 years of age. Studies substituting testosterone in elderly men with low serum testosterone have shown that men with clinical symptoms identical to the symptomatology of classical hypogonadism will benefit most from such therapy. Therefore, it is the general consensus to treat men with age-related hypogonadism only when clinical symptoms are present that can be potentially corrected by testosterone administration. Until recently, intramuscular injections of esters, such as testosterone enanthate, have been the mainstay of testosterone therapy. The introduction of testosterone patches has not challenged this approach, since many users of patches suffer from moderate to severe skin reactions. Some oral testosterone formulations have proven to be problematic, as absorption can be variable, bioavailability is frequently poor, due to the first-pass effect of the liver, and frequent administration is often required. Oral testosterone undecanoate avoids, at least partially, the first pass effect of the liver. However, plasma testosterone levels generally undergo large fluctuations. The large fluctuations in serum testosterone levels caused by conventional intramuscular injections result in unsatisfactory shifts in mood and sexual function in some men, which, combined with the frequency of injections, make the intramuscular mode of delivery far from ideal. Recently, a hydroalcoholic gel containing 1% testosterone has proven to be as efficient as a testosterone patch, but with fewer side-effects and a higher grade of patient satisfaction. Doses of 50-100 mg gel applied once daily on the skin deliver sufficient amounts of testosterone to restore normal hormonal values and correct the signs and symptoms of hypogonadism. The gel has been shown to be effective and successful in patients in the United States, who have benefited from its availability for almost 3 years. In the near future, intramuscular injections of testosterone undecanoate will become commercially available. Such injections have a very favorable pharmacokinetic profile, with one injection every 3 months maintaining serum testosterone well within the normal range. In phase III studies, intramuscular testosterone undecanoate proved to be as efficient as testosterone enanthate, with only one-quarter of the number of injections required and more stable serum testosterone levels. Thus, the new application modes--hydroalcoholic gel (for example, Testogel, Schering AG, Germany) and intramuscular testosterone undecanoate (Nebido, Schering AG, Germany)--appear to be the methods of choice in the near future, one being very suitable for hormone therapy in elderly men, the other for long-term substitution in classical forms of hypogonadism. PMID- 15799130 TI - Dyslexia. PMID- 15799129 TI - Androgen supplementation in elderly males: is dihydrotestosterone to be preferred? PMID- 15799131 TI - Neurobehavioural outcomes of mild head injury in children and adolescents. AB - The vast majority of closed-head injuries (CHI) in children are of mild severity. Even if only a small proportion of children with mild CHI suffer persistent negative outcomes, then mild CHI is a serious public health problem. This paper summarizes the existing literature regarding the neurobehavioural outcomes associated with mild CHI in children and adolescents, focusing on the longstanding debate regarding post-concussive syndrome. The paper also discusses conceptual and methodological issues that arise in research on the outcomes of mild CHI, including the definition of mild CHI; selection of comparison groups; measurement of outcomes; assessment of risk factors; timing of outcome assessments; and prediction of outcomes for individual children. The paper describes an ongoing research project that may help to resolve some of the longstanding controversies and uncertainties regarding the outcomes of mild CHI in children and concludes with a review of likely future directions for research on the outcomes of mild CHI. The long-term goal of research in this area is to develop a comprehensive and integrated biopsychosocial model of outcomes that can help guide clinical management. PMID- 15799132 TI - Adult life with Duchenne muscular dystrophy: observations among an emerging and unforeseen patient population. AB - The knowledge of adult life with Duchenne muscular dystrophy (DMD) is sparse. The purpose of this study was to review existing information and describe body functional, social participatory and quality of life profiles of the ordinary adult Danish DMD patient. Sixty-five study subjects aged 18-42 years were included in a cross-sectional survey based on data from a semi-structured questionnaire comprising 197 items. The ordinary adult DMD patient states his quality of life as excellent; he is worried neither about his disease nor about the future. His assessment of income, hours of personal assistance, housing, years spent in school and ability to participate in desired activities are positive. Despite heavy immobilization, he is still capable of functioning in a variety of activities that are associated with normal life. He lacks qualifying education and he is in painful need of a love life. The frequency of pains is surprisingly high; nearly 40% has pains daily. The nature, magnitude, consequence and possible cure of these reported pains must be scrutinized. Parents and professionals, paediatricians not the least, must anticipate in all measures taken that the DMD boy grows up to manhood and will need competences for adult social life in all respects. PMID- 15799133 TI - Interventions for post-traumatic stress disorder in children and adolescents, Alan Carr: clinical implications. PMID- 15799134 TI - Is rehabilitation the Cinderella of health, education and social services for children? PMID- 15799136 TI - Potential benefits of physical activity for children with acute lymphoblastic leukaemia. AB - Acute lymphoblastic leukaemia (ALL) is the most common form of paediatric leukaemia. The survival rate in children with ALL has improved significantly over the past several years, which makes quality of life an important focus for researchers. Some of the side effects of treatment (i.e. osteoporosis and obesity) are not realized until years after conclusion of therapy. Few studies have addressed the impact of physical activity (PA) on the side effects that occur during treatment of children with ALL. This paper discusses the increased risk for both osteoporosis and obesity due to treatment for ALL and suggests ways that PA may attenuate bone loss and risk of obesity by discussing what is known about effects of PA in healthy children and children with other chronic diseases. Recommendations will be made for PA interventions and future research in children with ALL. PMID- 15799135 TI - Lower extremity sensory function in children with cerebral palsy. AB - OBJECTIVES: (1) To determine the feasibility of qualitative sensory testing in the lower extremities (LE) of children with cerebral palsy (CP), especially spastic diplegia. (2) To determine if there is a detectable difference in qualitative LE sensory function in children with CP compared to typical children. (3) To determine if dorsal rhizotomy results in detectable changes in LE sensory function in children with spastic diplegia. DESIGN: Objectives 1 and 2: Prospective observational cohort study. Objective 3: Add-on to prospective interventional studies. SETTING: Regional tertiary children's hospital. PARTICIPANTS: Objectives 1 and 2: 62 children with CP and 65 typical children between 3-18 years of age. Objective 3: 34 children with spastic diplegia. INTERVENTIONS: Objectives 1 and 2: None. Objective 3: Dorsal rhizotomy. MAIN OUTCOME MEASURES: Pain, light touch, direction of scratch, vibration, toe position and knee position using standard qualitative techniques. RESULTS: Objective 1: 32 (52%) children with CP and 55 (85%) typical children completed all items (p = 0.09). Objective 2: Summary scores for separate LE sensory modalities were lower in children with CP for direction of scratch (p < 0.001), toe position (p = 0.01) and vibration sense (p = 0.01). Objective 3: No changes of LE sensory function. CONCLUSIONS: LE sensory testing in young children with CP is feasible. There is a qualitative sensory deficit in this sample of children with CP and specifically in children with spastic diplegia that is traditionally associated with dorsal column sensory modalities. A conservative dorsal rhizotomy does not produce a measurable change in LE sensory function in this sample of children with spastic diplegia. PMID- 15799137 TI - Micro-switch clusters to enhance hand responses and appropriate head position in two children with multiple disabilities. AB - This study evaluated the use of micro-switch clusters to improve response activity and posture with two children with multiple disabilities. The children were first taught a hand response and then required to combine this response with appropriate head position. The micro-switch clusters adopted for this purpose consisted of a pressure or mercury micro-switch for the hand response combined with a mercury micro-switch for the head position. Both children had an increase in the frequency of the hand response and in the percentage of times this occurred in combination with appropriate head position. These changes were maintained at a 2-month post-intervention check. PMID- 15799138 TI - Bedtime diazepam enhances well-being in children with spastic cerebral palsy. AB - In work with children with cerebral palsy at Ashirvad, Child Development and Research Centre, Chennai, India, the authors were confronted with fretful children who resisted any attempt to mobilize their limbs due to hypertonia and muscle spasm. It was found that administering a bedtime dose of diazepam to reduce hypertonia and muscle spasm alongside passive stretching exercises significantly improved the behaviour of the child. There was significant improvement in the well-being of the child during the activities of daily living and this reduced the family's burden of caring for the child. In this double blind, placebo-controlled, randomized clinical trial, each child received a bedtime dose of diazepam or placebo. The bedtime diazepam relaxed the muscles and this made the passive stretching easy and the movements sustained the muscle relaxation during the day. There were fewer unwarranted crying spells during the day and less wakefulness during the night. The adverse effect of day time sedation was not observed with the use of a single dose of diazepam at bedtime. PMID- 15799139 TI - Voices from the past: early institutional experience of children with disabilities--the case of Scotland. AB - In the countries of the western world, the institutional option for confining and endeavouring to 'correct' traits and attributes of people who did not conform to a rising consciousness of 'normalcy' developed rapidly during the 19th century. Scotland, a small, but rapidly urbanizing and industrializing country on the edge of Europe, was not an exception, but in its adoption of the institutional option for children with disabilities, its responses and objectives to different forms of impairment followed an uneven path. This article considers the reason for this erratic response, and the varied objectives of those responses, in the context of different forms of impairment and the differing interests of those with a professional or philanthropic involvement. The responses of some of the children directly affected are also considered. PMID- 15799140 TI - Spasticity: clinical perceptions, neurological realities and meaningful measurement. PMID- 15799141 TI - Clinical scales for the assessment of spasticity, associated phenomena, and function: a systematic review of the literature. AB - PURPOSE: To characterise clinical assessment methods for spasticity and/or its functional consequences in clinical patient populations at risk to suffer from spasticity. METHOD: Systematic literature search and manual-based two-step review process of psychometric properties of clinical assessment scales for spasticity and associated phenomena, as well as of functional scales with an association with spasticity. Reviewed psychometric properties included internal consistency, interrater, intrarater as well as retest reliability, construct validity, ecological validity, and responsiveness. RESULTS: Until May 2003 electronic database searches established a reference pool of 4151 references of which 90 references contributed to the review objectives. An additional 20 references were identified by an informal reference search. Twenty-four clinical scales that assess spasticity and/or related phenomena as well as 10 scales for 'active function' and three scales for 'passive function' with an association with spasticity could be identified. Some evidence signals that a high interrater reliability of the Ashworth and modified Ashworth scales can be achieved, however not in all circumstances. For many scales, reliability data is, however, missing. This is especially true for test retest reliability. Information about construct validity can promote our understanding of what individual scales are likely to assess. Many scales have been able to document changes after therapeutic intervention. CONCLUSIONS: The collated evidence can guide our clinical decision about when to use which scale and can promote evidence-based assessment of spasticity and related clinical phenomena. PMID- 15799142 TI - Biomechanical approaches applied to the lower and upper limb for the measurement of spasticity: a systematic review of the literature. AB - PURPOSE: To review and characterise biomechanical approaches for the measurement of spasticity as one component of the upper motor neurone syndrome. METHOD: Systematic literature searches based on defined constructs and a four-step review process of approaches used or described to measure spasticity, its association with function or associated phenomena. Most approaches were limited to individual joints and therefore, to reflect this trend, references were grouped according to which body joint(s) were investigated or whether it addressed a functional activity. For each joint, references were further sub-divided into the types of measurement method described. RESULTS: A database of 335 references was established for the review process. The knee, ankle and elbow joints were the most popular, perhaps reflecting the assumption that they are mono-planar in movement and therefore simpler to assess. Seven measurement methods were identified: five involving passive movement (manual, controlled displacement, controlled torque, gravitational and tendon tap) and two involving active movement (voluntary and functional). Generally, the equipment described was in an experimental stage and there was a lack of information on system properties, such as accuracy or reliability. Patient testing was either by cohort or case studies. The review also conveyed the myriad of interpretations of the concept of spasticity. CONCLUSIONS: Though biomechanical approaches provide quantitative data, the review highlighted several limitations that have prevented them being established as an appropriate method for clinical application to measure spasticity. PMID- 15799143 TI - Neurophysiological methods for the assessment of spasticity: the Hoffmann reflex, the tendon reflex, and the stretch reflex. AB - PURPOSE: To review the literature concerning neurophysiological methods to assess spasticity with respect to mechanisms and methodology, and to describe the three most commonly used methods: the Hoffmann reflex (H-reflex), the Tendon reflex (T reflex), and the Stretch Reflex (SR). METHOD: A systematic internet database search was performed to identify neurophysiological measurement methods of spasticity. A systematic exclusion procedure resulted in 185 included references, completed by additional informal search. For this paper, information about the H , T- and stretch reflexes was extracted from these references. RESULTS: Although the reflexes are basically monosynaptic, there are many supraspinal pathways which modulate the responses in terms of their amplitude and latency. As a consequence the methods are sensitive to a considerable number of experimental conditions and are characterized by a moderate reliability and sensitivity. Correlations with other (i.e. biomechanical, neurophysiological or clinical) spasticity assessment parameters are moderate to poor. Standardised and broadly accepted protocols are still largely lacking preventing an effective exchange of knowledge. CONCLUSIONS: The clinical and experimental use of the three methods is restricted due to moderate reliability and sensitivity. It is recommended to perform combined neurophysiological-biomechanical assessment of spasticity during active, functional movement. PMID- 15799145 TI - New proposals for training in neurosurgery. AB - The rapidly evolving changes in working patterns, career structure and the regulation of training of doctors have provided an ideal opportunity for proposals to improve the programme for the training of neurosurgeons. The Education and Training Committee of the Society of British Neurological Surgeons (D.G. Hardy, A. J. W. Steers, N. T. Gurusinghe, P. M. Foy, P. van Hille, R. A. Cowie, H. A. Crockard, O. Sparrow and S. Burn) has, in recent months, worked closely with the Specialist Advisory Committee (SAC) in neurosurgery, Department of Health (Modernizing Medical Careers Group, H. A. Crockard, A. Havers, T. Hobbs) and colleagues from the major neuroscience specialties to develop a new programme based on a 'Common Stem' approach. This article describes the principles of the programme. The proposals have received approval by the Council of the SBNS, the Presidents of the four Surgical Royal Colleges and the Senate of Surgery. PMID- 15799146 TI - A study of acute rehabilitation after head injury. AB - In 1999 a report from the Royal College of Surgeons of England drew attention to the fact that, in the UK, there is little structured acute inpatient rehabilitation for patients admitted after traumatic brain injury (TBI). This paper reports the results of a retrospective study of acute inpatient rehabilitation (AR) after TBI, comparing aspects of resource use in 92 patients who received structured unit-based multidisciplinary AR with 97 patients who received usual practice. About 10% of patients admitted via casualty after TBI required AR after neurosurgical consultation or care. These patients remained in AR for a mean of 20.5 days; length of stay in an acute bed was not prolonged compared with patients in usual practice. Sixty per cent of patients discharged home from unit-based care were referred to community-based rehabilitation on discharge compared with no patients discharged home from neurosurgical care. There is an urgent need to discuss and plan at national level the structured secondary provision of acute rehabilitation after acquired brain injury. PMID- 15799144 TI - Theoretical and methodological considerations in the measurement of spasticity. AB - PURPOSE: To discuss the measurement of spasticity in the clinical and research environments, make recommendations based on the SPASM reviews of biomechanical, neurophysiological and clinical methods of measuring spasticity and indicate future developments of measurement tools. METHOD: Using the results of the systematic reviews of the biomechanical, neurophysiological and clinical approaches, methods were evaluated across three dimensions: (1) validity, reliability and sensitivity to change; (2) practical quality such as ease of use and (3) qualities specific to the measurement of spasticity, for example ability to be applied to different muscle groups. Methods were considered in terms of applicability to research and clinical applications. RESULTS: A hierarchy of measurement approaches was identified from highly controlled and more objective (but unrelated to function) to ecologically valid, but less objective and subject to contamination from other variables. The lack of a precise definition of spasticity may account for the problem of developing a valid, reliable and sensitive method of measurement. The reviews have identified that some tests measure spasticity per se, some phenomena associated with spasticity or consequential to it and others the effect of spasticity on activity and participation and independence. CONCLUSIONS: Methods appropriate for use in research, particularly into the mechanism of spasticity did not satisfy the needs of the clinician and the need for an objective but clinically applicable tool was identified. A clinical assessment may need to generate more than one 'value' and should include evaluation of other components of the upper motor neurone syndrome. There is therefore a need for standardized protocols for 'best practice' in application of spasticity measurement tools and scales. PMID- 15799147 TI - Posterolateral thoracic disc disease: clinical presentation and surgical experience with a modified approach. AB - Posterolateral thoracic disc disease is often misdiagnosed and labelled as demyelination. A high index of suspicion and awareness of this disease entity is the key to diagnosis and treatment. Our experience with a modified posterolateral facetal-sparing pedicle-sparing approach is highlighted. PMID- 15799148 TI - Association between dynamic cerebral autoregulation and mortality in severe head injury. AB - The objective of the study was to test the hypothesis that dynamic cerebral pressure-autoregulation is associated with the outcome of patients with severe head injury and to derive optimal criteria for future studies on the predictive value of autoregulation indices. Repeated measurements were performed on 32 patients with severe head injury. Arterial blood pressure (ABP) was measured continuously with an intravascular catheter, intracranial pressure (ICP) was recorded with a subdural semiconductor transducer and cerebral blood flow velocity (CBFV) was measured with Doppler ultrasound in the middle cerebral artery. Transfer function analysis was performed on mean beat-to-beat values, using ABP or CBFV as input variables and CBFV or ICP as the output variables. A dynamic index of autoregulation (ARI) ranging between 0 and 9 was extracted from the CBFV step response for a change in ABP. No significant differences between survivors and non-survivors were found due to mean values of ICP, ABP, CPP, CBFV, pCO2, GCS, age or heart rate. The transfer functions between ABP-ICP and CBFV-ICP did not show any significant differences either. The median [lower, upper quartiles] ARI was significantly lower for non-survivors compared with survivors [4.8 (0.0, 5.9) v. 6.9 (5.9, 7.4), p= 0.004]. The correlation between ARI and GOS was also significant (r=0.464, p=0.011). Cohen's coefficient was optimal for a threshold of ARI= 5.86 (kappa 0.51, p=0.0036), leading to a sensitivity for death of 75%, specificity=76.5%, odds ratio =9.75 and overall precision = 75.8%. The difference in ARI values between survivors and non-survivors persisted when results were adjusted for GCS (p = 0.028). A similar analysis for the Marshall CT scale did not reach significance (p = 0.072). A logistic regression analysis confirmed that apart from the ARI, no other variables had a significant contribution to predict outcome. In this group of patients, death following severe head injury could not be explained by traditional indices of risk, but was strongly correlated to indices of dynamic cerebral pressure-autoregulation extracted by means of transfer function analysis. Future studies using a prospective design are needed to validate the predictive value of the ARI index, as estimated by transfer function analysis, in relation to death and other unfavourable outcomes. PMID- 15799149 TI - Reliability of diagnosis of soft cervical disc prolapse using Spurling's test. AB - The purpose of this study was to determine the sensitivity and specificity of the Spurling's test in predicting the diagnosis of a soft lateral cervical disc prolapse. A prospective study was performed involving 50 patients with neck and arm pain; of these, 25 patients were managed surgically (Group 1) and the other 25 patients were managed conservatively (Group 2). A Spurling's test was performed in all patients and scored as positive or negative. In patients in Group 1, the operative findings were recorded as a soft or hard disc prolapse (osteophyte). In Group 2 patients, the MR findings were recorded as positive for a soft disc prolapse or negative (no disc prolapse or osteophyte only). The results of the Spurling's test were compared with the surgical or MR findings, which were considered as the gold standard. Spurling's test had a sensitivity of 92% and specificity of 95% with positive predictive value (PPV) of 96.4% and negative predictive value (NPV) of 90.9% in predicting a diagnosis of a soft lateral cervical disc prolapse. The high positive predictive value of the test can be utilized to improve the yield of positive MR examinations in patients with cervical radiculopathy. PMID- 15799150 TI - Extradural arachnoid cysts: a study of seven cases. AB - Analysis of the management of 7 cases of extradural arachnoid cyst is presented. A brief review of the literature on the subject is presented. Seven cases of extradural arachnoid cysts were treated surgically between the years 1996 to 2002. These patients were analysed retrospectively. The average follow-up period was 32 months. Three patients were males and four were females and their ages ranged from 12 to 35 years. The patients presented with predominant complaint of progressive motor deficits. There were two cysts in the dorsal region, four in the dorsolumbar region and one in the cervicodorsal region. The extradural arachnoid cyst was located posterior to the dural tube and in the midline in six cases and all these cysts had bilateral multiple intervertebral foraminal extensions. In the case with a cervicodorsal cyst, the cyst was anterolateral to the dural tube and there was no foraminal extension. The surgery comprised of wide exposure of the cyst and radical marsupialization or resection of the cyst wall. There was no recurrence of the cyst during the period of follow-up. Radical resection of the walls leads to a lasting cure from extradural arachnoid cyst. The site of communication of the cyst contents with subarachnoid CSF could not be identified during the surgery in the series. PMID- 15799151 TI - Idiopathic granulomatous hypophysitis presenting as non-functioning pituitary adenoma: description of six cases and review of literature. AB - The aim of this study was to analyse the complete profile and outcome of patients with idiopathic giant cell granulomatous hypophysitis. Six consecutive cases of idiopathic giant cell granulomatous hypophysitis were studied from 1993 to 2002. Headache and visual disturbances were the most frequent presenting symptoms. All patients had hypogonadism, four had hypoadrenalism and three were hypothyroid at presentation. None of them had diabetes insipidus preoperatively. A sellar mass with suprasellar extension on MRI with loss of the posterior pituitary 'bright spot' was a consistent observation in all patients. All patients underwent surgical excision of the mass lesion with histopathological confirmation of giant cell granulomatous hypophysitis. Other systemic granulomatous diseases were excluded by appropriate investigations. Postoperatively, all patients became hypothyroid and hypogonad, five patients had adrenal insufficiency, while two developed permanent diabetes insipidus. The clinical presentation of giant cell granulomatous hypophysitis is that of an expanding sellar mass lesion with a varying degree of endocrine dysfunction. Preoperative diagnosis of 'hypophysitis' is usually difficult; however, stalk thickening and loss of posterior pituitary 'bright spot' on MR imaging are clues to the diagnosis. PMID- 15799152 TI - Hydroxyurea chemotherapy for meningiomas: enlarged cohort with extended follow up. AB - Meningiomas account for 18-20% of all intracranial tumours and often recur despite surgical resection. Hydroxyurea is under evaluation as adjuvant therapy of meningiomas. In the authors' initial report of 17 patients with meningioma, hydroxyurea demonstrated modest efficacy, with a median time to progression (TTP) of 80 weeks. In the current study, 21 patients with meningioma have been placed on hydroxyurea (20 mg/kg/day orally), with extended follow-up of the original cohort. Eighteen of 20 evaluable patients (90%) responded with stable disease ranging from 20 to 328 + weeks (median TTP 176 weeks; 11 patients censored). Five of the stabilized patients progressed after 20, 56, 36, 216 and 56 weeks, respectively. Two patients had progressive disease after 10 weeks. Toxicity was mainly haematological. Hydroxyurea has modest activity against meningiomas and should be considered for patients who are poor surgical candidates, have unresectable or large residual meningiomas, or have progressed after surgical resection or irradiation, or both. PMID- 15799153 TI - Dramatic response of malignant craniopharyngioma to cis-platin-based chemotherapy. Should craniopharyngioma be considered as a suprasellar 'germ cell' tumour? AB - The case is described of a 21-year-old woman, who developed a malignant tumour arising from a craniopharyngioma 14 years after the original diagnosis. The remarkable response of this malignant tumour ex-craniopharyngioma to cis-platin based chemotherapy, together with other midline tumour characteristics of craniopharyngioma, raise the question as to whether craniopharyngioma should any longer be separately considered from suprasellar germ cell tumour. This subject is discussed. PMID- 15799154 TI - A 25-gauge needle used as an arachnoid knife in microneurosurgery. PMID- 15799156 TI - Multicystic, heterogeneously enhancing meningioma in an octogenerian. AB - A multicystic meningioma in an octogenerian whose tissue diagnosis was ill defined and misleading on preoperative neuroradiologic imaging is presented. Nauta has described four cyst types that can develop in cystic meningiomas. We report the first case in which three cyst types are demonstrated concurrently, describe the histopathology and surgical management. This case represents a rare variant of a common tumour in an unusual age group, and underscores the need for definitive biopsy and resection as indicated. Furthermore, the diagnosis of multicystic meningioma does not favour an aggressive histopathology in this case. PMID- 15799157 TI - Idiopathic chronic hypertrophic pachymeningitis causing thoracic cord compression. AB - Chronic idiopathic pachymeningitis is a rare cause of compressive myelopathy, especially of the thoracic spinal cord. The hypertrophic dura causes symptoms either by pressure on the spinal cord or by vascular compromise. The authors report the diagnostic dilemmas in a 48-year-old lady who presented with progressive spastic paraparesis of 4 months duration. MRI showed an extradural hypointense band, surrounding and flattening the thoracic cord in the anteroposterior direction, extending from C7 to T11. Neither the CT scan nor the myelogram provided any clue to the diagnosis. A T1-T10 laminectomy was performed. The dura was hypertrophied and found compressing the cord. The posterior dura was excised and the spinal cord was decompressed. The histopathology showed hypertrophic pachymeningitis. The patient improved neurologically. She was started empirically on anti-tubercular chemotherapy, though she was serologically negative for all known causes of pachymeningitis. PMID- 15799158 TI - Diffuse primary non-Hodgkin's lymphoma of the cranial vault. AB - Primary non-Hodgkin's lymphoma of the skull vault is a rare disease. We describe a case occurring in a 72-year-old woman presenting with generalized tonic clonic seizures on a background of a 1-year history of headaches and progressively enlarging scalp masses. Imaging showed diffuse infiltration of the skull vault with multifocal intra- and extracranial soft tissue masses, causing compression and probably infiltration of the cerebral cortex. Further investigation failed to identify any other evidence of systemic lymphoma. Biopsy of one of the scalp masses showed a small to intermediate cell B cell lymphoma. The other nine reported cases of primary skull vault lymphoma are reviewed. The diffuse vault infiltration as well as the multiple intracranial, scalp and temporalis muscle masses renders this case unique. PMID- 15799155 TI - Complications after trans-sphenoidal surgery: our experience and a review of the literature. AB - The incidence of surgical complications after trans-sphenoidal surgery for pituitary lesions is low. The influence of size of the lesion and its pathology on the incidence of different types of complications and the remission rate of functioning adenomas are addressed in this retrospective study. Between 1996 and 2001, 126 trans-sphenoidal operations were performed on 108 patients with pituitary pathologies. Diabetes insipidus (DI) was the commonest (23%) and the incidence was higher with microadenomas and in those with Cushing's disease (p < 0.05). Other important complications were: postoperative hypopituitarism 22%, CSF leak 13%, meningitis 5.5%, pneumocephalus 2.4%, visual deterioration 1.5%, haematoma 0.8% and 30-day mortality rate of 0.8%. Other complications encountered were subdural haematoma, epistaxis, SIADH, sphenoid abscess, deep vein thrombosis, thalamic infarct and hydrocephalus. No vascular injuries were encountered. The overall remission rate for acromegaly and Cushing's disease (22 patients each) was 77 and 81%, respectively. The remission rate was 100% for microadenomas, 89% for both acromegaly and Cushing's disease. More aggressiveness towards complete tumour removal increases the biochemical cure rate of functioning adenomas and on the other hand results in higher incidence of CSF leak and hypopituitarism. PMID- 15799159 TI - Pregnancy related pituitary enlargement mimicking macroadenoma. AB - A 38-year-old woman presented at 30 weeks gestation with bi-temporal upper quadrantanopia. MRI showed appearances typical of a pituitary macroadenoma. It was agreed that pituitary decompresssion would only be indicated if the visual field defect worsened. MRI imaging 2 weeks postdelivery showed complete resolution of the pituitary lesion. PMID- 15799160 TI - Fibrous dysplasia of the cervical spine presenting as a pathological fracture. AB - Fibrous dysplasia (FD) is an uncommon benign fibro-osseous abnormality of bone, of unknown aetiology and equal sex incidence, which is most commonly restricted to one bone (monostotic FD: MFD, 70%). Less commonly multiple bones are affected (polyostotic FD: PFD, 27%). Vertebral involvement is uncommon (4%), but more common with PFD (7 - 24%) than MFD (1%). Of 20 cases of FD involving the cervical spine, only three have represented MFD. Unlike cases associated with PFD, all cases presented with acute neck pain without significant neurological impairment after minor trauma. We present the case of a 35-year-old male with MFD who developed a pathological fracture of C3 following minor trauma. Radiographs showed collapse and typical 'ground glass' lucency of C3. CT revealed replacement of C3 cancellous bone by hypodense tissue extending into the right lateral mass. The cortex was thinned and fractured, and encroached upon the right foramen transversarium and spinal canal. Magnetic resonance imaging demonstrated hypo intensity on both T1 and T2, with uniform contrast enhancement. Subtotal excision was achieved via an anterior C3 corpectomy, with residual FD left within the right lateral mass. Stability was achieved utilizing an iliac crest strut autograft, C2-4 plate-and-screws, and mobilization in a halo frame for 3 months. At 18 months, he remained asymptomatic and without deficit. Radiography, CT and MRI confirmed graft fusion without FD invasion, but with residual right lateral mass FD unchanged in size. PMID- 15799161 TI - Complex neurobehavioural syndrome due to bilateral thalamic glioma. AB - We present a 65-year-old female with bilateral thalamic astrocytoma. The unusual long survival of this patient allowed the manifestation of a complex neurobehavioural syndrome due to gradual involvement of several thalamic nuclei. An attempt is made to approach the complexity of symptoms according to the anatomical areas, nuclei and thalamic connections infiltrated. PMID- 15799162 TI - Bone wax can cause foreign body granuloma in the medulla oblongata. AB - Bone wax is commonly used in neurosurgical practice as a haemostatic and sealing agent. It is a safe agent, but may lead to adverse effects such as infection, epistaxsis, allergic reaction or foreign body granuloma. There are very few reported cases of the neurological complications of remnant bone wax in the subdural or subarachnoid space. The authors report the case of a foreign body granuloma infiltration through the medulla oblongata due to remnant bone wax in the subarachnoid space after posterior fossa decompressive surgery. PMID- 15799163 TI - Improvement in cognitive function after right temporal arteriovenous malformation excision. AB - A patient is presented in whom right temporal AVM excision led to improved cognitive function. Neuropsychological assessment showed a significant postoperative improvement in Performance IQ (p = < 0.05), Full Scale IQ (p = < 0.05), and clinically in visual memory (p = < 0.10). Reversal of vascular steal may lead to increases in cognitive function. We recommend that patients undergoing AVM excision have a complete neuropsychological evaluation. PMID- 15799164 TI - Stab wound of the cervical spinal cord and ipsilateral vertebral artery injury. AB - Stab wound of the cervical spinal cord with ipsilateral vertebral artery injury is rare. We report a case of a penetrating injury by scissors to the cervical spinal cord and right vertebral artery. The management is discussed and literature reviewed. PMID- 15799166 TI - Multiple brain metastases from malignant melanoma with long-term survival. AB - We present a case of multiple malignant melanoma metastases in the brain who is leading a normal life 16 years after the brain secondaries were managed by surgical resection, stereotactic radiation and chemotherapy. The primary lesion in the left upper arm was excised 4 years prior to the brain metastases. His most recent MRI shows him to be disease free. To the best of our knowledge, this is longest survival reported of any patient with multiple brain metastases from malignant melanoma. PMID- 15799165 TI - Intramedullary subependymoma of the lower spinal cord. AB - A 22-year-old woman developed an increasing distal paraparesis, which resulted in a severe ataxic gait. MRI demonstrated a very large intramedullary tumour mass from D11 to L2, which could not be identified as one of the common neoplastic diseases of the spinal cord. The tumour was removed completely and the diagnosis of subependymoma confirmed. With respect to our patient and to the limited reports in the literature, we describe some typical features that may help in the diagnostic and preoperative assessment of this very rare spinal cord tumour. PMID- 15799167 TI - Posterior-fossa intra-tumoural abscess: a report of three patients and literature review. AB - Intraparenchymal posterior fossa neoplasms containing intratumoural abscesses are rare. Intrasellar or parasellar tumours are among the common neoplasms developing such a complication, as the result of direct extension of the microbial flora from the contiguous infected sinuses. All meningiomas reported with such a complication have been located parasagittally. A short series of three posterior fossa tumours, containing intra-tumoural abscesses, are described, of which only one was diagnosed pre-operatively. A haematogenous pathway was considered as the mode of spread in all three patients. Magnetic resonance imaging was performed in all patients and the features are discussed. PMID- 15799168 TI - Supratentorial intraventricular schwannoma. AB - A rare case of supratentorial intraventricular schwannoma is described. A 16-year old male with asymptomatic papilloedema was found to have a 5-cm intraventricular tumour arising from the trigone of the right lateral ventricle. A craniotomy and total macroscopic excision was performed. PMID- 15799169 TI - The case report of Al-Anazi et al. PMID- 15799170 TI - Changes in serum interleukin-6 and high-sensitivity C-reactive protein levels in patients with acute coronary syndrome and their responses to simvastatin. AB - The role of inflammation in acute coronary syndrome (ACS) and the mechanism by which statin treats ACS is explored. Serum high-sensitivity C-reactive protein (hs-CRP) and interleukin-6 (IL-6) levels were measured in 50 patients with ACS [including 30 cases with unstable angina (UA) and 20 patients with acute myocardial infarction (AMI)], 34 patients with stable angina (SA), and 30 controls. Patients in the ACS group were randomly assigned to a simvastatin group (including a simvastatin AMI subgroup, n = 11 and a simvastatin UA subgroup, n = 14) and a routine group (including a routine AMI subgroup, n = 9 and a routine UA subgroup, n = 16). The simvastatin group was given simvastatin 20mg/day and the routine group a placebo. After a 3-week follow-up, serum hs-CRP, IL-6 levels, and serum lipid concentrations were measured again. Both serum IL-6 and hs-CRP levels were significantly higher in the ACS group (including the UA and AMI subgroups) than in the SA and control groups (P < 0.001). After 3 weeks of treatment with simvastatin, the serum IL-6, hs-CRP, total cholesterol, and low-density lipoprotein cholesterol levels were decreased significantly in the simvastatin group (P < 0.001), but no significant changes were observed in the routine group. No relationship was observed between the rate of decrease of serum IL-6 or hs-CRP and serum lipids levels. The hs-CRP level showed a significant correlation with IL-6 by Spearman's rank correlation analysis (P < 0.01). Inflammation plays an important role in the initiation of ACS. Simvastatin possesses an anti inflammatory effect, independent of its lipid-lowering action, which may play an important role in the early treatment of ACS. PMID- 15799171 TI - Effects of interferon-alpha therapy on cardiac function in patients with chronic hepatitis B infection. AB - Various types of cardiovascular complications, such as myocardial infarction, cardiac arrhythmias, cardiomyopathy, and myocarditis attributed to interferon therapy have been reported. The aim of this study was to evaluate the cardiac effects of interferon-alpha (IFN-alpha) in patients with chronic hepatitis B infection. Forty-five patients with chronic hepatitis B infection (41 men and 3 women; mean age 34.2 +/- 11.5 years) were included in the study, and 10MU IFN alpha 2b was administered three times a week for 6 months to the patients. Cardiac evaluation (detailed medical history, physical examination, electrocardiography, systolic and diastolic function parameters by echocardiography) was performed at the beginning, and at the 1st, 3rd, and 6th months of therapy. No patients had any cardiac symptoms during interferon therapy, and systolic and diastolic blood pressure and heart rate were not significantly affected (P > 0.05). None of the patients revealed cardiac rhythm disturbance on electrocardiography before or during the therapy period. No significant changes were detected in systolic (ejection fraction, fractional shortening, pre-ejection period, left ventricular ejection time, the ratio of pre ejection period/ejection time, Q-V peak) and diastolic (E peak, A peak of transmitral flow velocity, E/A ratio, deceleration time, isovolumic relaxation time by conventional echocardiography, and E peak, A peak, deceleration time of E wave at the medial and lateral corners of the mitral annulus by tissue Doppler echocardiography) left ventricular function parameters between the beginning and the 1st, 3rd, and 6th months of therapy. The results of this study suggest that IFN-alpha therapy does not cause a significant deterioration in cardiac function in patients with chronic hepatitis B infection, and it may be used safely in patients without cardiac disease. PMID- 15799173 TI - Documentation of slow coronary flow by the thrombolysis in myocardial infarction frame count in habitual smokers with angiographically normal coronary arteries. AB - The thrombolysis in myocardial infarction (TIMI) frame count is a simple clinical tool for assessing quantitative indexes of coronary blood flow. In this study we aimed to evaluate the effects of long-term cigarette smoking on the TIMI frame count in patients with angiographically proven normal coronary arteries. Between May 2001 and January 2002, 41 habitual smokers and 41 sex-matched nonsmokers with angiographically proven normal coronary arteries were included in the study. The TIMI frame count was determined for each major coronary artery in each patient. The TIMI frame count of the smoking group was significantly higher than that of nonsmokers for all three coronary arteries: left anterior descending (corrected), 39 +/- 13 vs 22 +/- 8; right coronary artery, 35 +/- 13 vs 24 +/- 11; and left circumflex artery, 37 +/- 13 vs 25 +/- 8 (P < 0.001 for all). The smokers tended to be younger than nonsmokers (46 +/- 7 vs 49 +/- 9 years; P = 0.07). We have found that smokers with angiographically normal coronary arteries have a higher TIMI frame count than nonsmokers with angiographically normal coronary arteries. An increased TIMI frame count can be regarded as an index of the harmful effects of smoking on coronary circulation regardless of the underlying mechanism. PMID- 15799172 TI - Factors affecting the blood concentration of ionized magnesium in patients in the acute phase of myocardial infarction. AB - Magnesium is physiologically active in its free state (Mg2+). In the present study, we attempted to clarify factors affecting blood concentrations of Mg2+ in the acute phase of myocardial infarction (AMI). Subjects were 84 consecutive patients with AMI. Blood samples were collected at the time of admission, 24h after admission, and 1 week after admission, to measure blood concentration of Mg2+ and noradrenaline (NA). Furthermore, to assess daily Mg intake the hardness of local drinking water was determined, and a survey was conducted regarding dietary preferences and habits. Based on the results of this survey, the patients were defined as having a low Mg intake (L Group) or not. In addition, based on chronological shifts in blood Mg2+ concentrations, subjects were divided into the following four groups: Normal group, blood concentration of Mg2+ within normal range at all measurement points; Early recovery group, low at time of admission, but normalized on the next day; Delayed recovery group, low at time of admission, but normalized 1 week after admission; and Unrecovered group, below normal range at all measurement points. The mean blood Mg2+ concentration on admission was 0.52 +/- 0.06 mmol/l, significantly lower than the normal range (P < 0.05). A negative correlation between blood Mg2+ and NA concentrations on admission was observed (r = 0.49, P < 0.005). As a result, blood Mg2+ concentrations were normalized in 94% of subjects by 1 week after admission. Mean blood Mg2+ concentration on admission in the L Group was 0.47 +/- 0.05 mmol/l, significantly lower than that found in other subjects (0.52 +/- 0.05 mmol/l, P < 0.01). Eighty percent of the patients classified into the Unrecovered group belonged to the L Group. These findings suggest that lower blood concentrations of Mg2+ and higher plasma NA levels may be a result of serious AMI. However, chronic Mg intake deficiency may play a partial role in patients whose blood concentrations of Mg2+ remain low for long periods of time. PMID- 15799174 TI - The prevalence and clinical predictors of atherosclerotic renal artery stenosis in patients undergoing coronary angiography. AB - Renal artery stenosis is an important cause of secondary hypertension as well as ischemic nephropathy. The purpose of this study was to determine the clinical predictors in patients with renal artery stenosis in a population referred for coronary angiography. From March 1998 to July 1999, 1459 patients undergoing coronary angiography for various indications were routinely screened for renal artery stenosis by undergoing abdominal aortography. Coronary angiography, carotid angiography, and abdominal aortography was performed via either the radial or the femoral approach. The data were analyzed retrospectively. Out of 1459 patients undergoing abdominal aortography, 158 (10.8%) were found to have significant renal artery stenosis with 24 of the patients having bilateral stenosis. Significant coronary artery diseases were found in 994 of the 1459 study population (68.1%), with 134 (13.5%) of these patients having concomitant renal artery stenosis. Multivariate logistic regression showed that extracranial carotid artery stenosis odds ratio [(OR) 4.89 (95% confidence interval 2.57 9.33), P < 0.001], peripheral artery disease [OR 4.64 (2.65-9.33), P < 0.001], renal insufficiency [OR 2.68 (1.43-5.02), P = 0.002], significant coronary artery disease [OR 2.01 (1.12-3.59), P = 0.019], hypercholesterolemia [OR 1.92 (1.07 3.43), P = 0.028], hypertension [OR 1.85 (1.16-2.95), P = 0.010], and old age (> 60 years) [OR 1.64 (1.01-2.64), P = 0.044] were significant clinical predictors of renovascular disease. The prevalence of indolent atherosclerotic renal artery stenosis is relatively high in selected groups of patients with high clincial risk factors for this underdiagnosed disease. Renal artery stenosis should be highly suspected in patients who have these risk factors because early detection of this disease may reverse the progression to chronic renal failure and end stage renal disease. PMID- 15799176 TI - Angiotensin-converting enzyme gene polymorphism, left ventricular remodeling, and exercise capacity in strength-trained athletes. AB - The mechanisms that regulate the development of human physiological cardiac hypertrophy remain poorly understood. The renin-angiotensin system, which is modulated by genetic polymorphism, plays an important role in the regulation of vascular tone and myocardial hypertrophy. Although a few studies have analyzed the association of angiotensin-converting enzyme (ACE) polymorphism and left ventricular (LV) hypertrophy in isotonic exercise-trained subjects who developed eccentric cardiac hypertrophy, there has been no research done in power athletes who developed concentric cardiac hypertrophy. We have hypothesized that ACE genotypic modulation characteristics may affect LV mass in power athletes. This study included 29 elite Caucasian wrestlers (mean age, 22.6 years) and 51 age matched sedentary subjects. According to the absence or presence of the insertion segment in the polymerase chain reaction (PCR) product, the subjects were classified as homozygous deletion-deletion (DD), insertion-insertion (II), or heterozygous insertion-deletion (ID). The association of LV hypertrophy with ACE gene insertion/deletion (I/D) polymorphism was analyzed. Left ventricular mass and index were determined by echocardiography. Angiotensin-converting enzyme genotyping was performed on peripheral leukocytes using the polymerase chain reaction technique. The study and control group subjects were similar in height and weight. Left ventricular hypertrophy in the athletes was more apparent than in the controls. Angiotensin-converting enzyme genotype II frequency was 17.2% (5) in the athletes, 17.6% (9) in the controls; ID frequency was 51.7% (15) in the athletes, 56.8% (29) in the controls; and the DD frequency was 31% (9) in the athletes and 25.4% (13) in the controls. Left ventricular mass and mass index were found to be higher in genotype DD (126.2 +/- 2.9g/m2) than genotype II (85.5 +/- 4.0g/m2) or genotype ID (110.1 +/- 2.3g/m2) in the athletes (P < 0.001). Furthermore, maximal oxygen consumption in genotype DD was found to be higher than in II and ID. An association was found between ACE gene I/D polymorphism and LV hypertrophy in strength-trained athletes. PMID- 15799175 TI - The association between coffee consumption and plasma total homocysteine levels: the "ATTICA" study. AB - The aim of this work was to investigate the association between homocysteine levels and coffee consumption in a sample of cardiovascular disease-free men and women. From May 2001 to December 2002, we randomly enrolled 1514 men and 1528 women, stratified by age and gender, from the greater area of Athens. Blood samples were collected in the fasting state. Among other investigated factors, dietary habits (including coffee consumption in ml per day, adjusted for 28% caffeine containment) were evaluated using a validated food frequency questionnaire. Men consumed higher quantities of coffee compared with women (250 +/- 55 vs 150 +/- 60 ml/day, P = 0.001), while homocysteine values were also higher in men than in women (14.5 +/- 6 vs 10.8 +/- 3.5 micromol/l, P = 0.001). A dose-response relationship of homocysteine levels with coffee consumption was observed (r = 0.10, P = 0.034). In particular, we found that homocysteine levels were 11.2 +/- 5 micromol/l for no consumption, 11.7 +/- 7 micromol/l for <100 ml/day, 12.5 +/- 7 micromol/l for 200-400 ml/day, and 12.7 +/- 4 micromol/l for >500 ml/day consumption (P = 0.018). The observed trend remained significant even after controlling for the interactions between coffee consumption with gender, smoking habits, physical activity status, and eating habits. However, the sole effect of the consumption of filtered coffee on homocysteine levels was significant only in those who consumed more than 500 ml/day (P = 0.043). Although our findings cannot be evidence for causality, they can be the basis for hypotheses about the relation between homocysteine and coffee that can partially explain the mechanisms by which elevated homocysteine levels may influence coronary risk. PMID- 15799178 TI - Assessment of coronary artery abnormalities in a patient with Kawasaki disease by multislice computed tomography. AB - The high spatial resolution of multislice computed tomography (MSCT) permits direct visualization of the coronary artery system. In this report, we describe coronary artery abnormalities in a young adult with Kawasaki disease. MSCT detected a giant coronary artery aneurysm, coronary artery stenosis in the first diagonal artery, and a multi-layered structure in the right coronary artery and the left circumflex artery. These findings corresponded well to those obtained by coronary angiography. MSCT has the potential to be the standard diagnostic tool for the follow-up evaluation of coronary artery disease in adolescents and young adults with Kawasaki disease. PMID- 15799177 TI - The first spontaneous coronary artery perforation due to disruption of atherosclerotic plaque. AB - Spontaneous coronary artery rupture is rarely seen and most of the reorted cases in the world literature are related to rupture of a coronary artery aneurysm or of a saphenous vein graft. There is no report in the literature of a patient with spontaneous coronary artery perforation due to disruption of coronary atherosclerotic plaque. We can confirm that our patient is the first to be successfully treated with intracoronary grafted stent implantation for spontaneous coronary artery perforation as a result of disruption of atherosclerotic plaque. PMID- 15799179 TI - Factors associated with delivery complications in rural Bangladesh. AB - OBJECTIVES: There are only a few studies on maternal morbidity, delivery complications and maternal mortality in Bangladesh. This study analyzes data from a follow-up study conducted by the Bangladesh Institute of Research for Health and Technologies (BIRPERHT) on maternal morbidity in rural Bangladesh in 1993. METHODS: A total of 1020 pregnant women were interviewed in the follow-up component of the study. The survey collected information on socioeconomic and demographic characteristics, pregnancy-related care and practice, morbidity during the period of follow-up as well as in the past, information concerning complications at the time of delivery and during the postpartum period. For the purpose of this study, we selected 993 pregnant women with at least one antenatal follow-up. Both bivariate and multivariate analyses were conducted to identify the potential risk factors for complication during delivery and duration of labor. RESULTS AND CONCLUSIONS: It appears that complications during the antenatal period can result in various complications at the time of delivery. Some of the important findings are: hemorrhage during the antenatal period increases the risk of excessive hemorrhage during delivery, the risk of obstructed labor increases significantly if abdominal pain is observed during the antenatal period, prolonged labor appears to be significantly higher for the first pregnancy, and pregnancies suffering from abdominal pain during pregnancy tend to have a higher risk of prolonged labor during delivery. The duration of labor appears to be negatively associated with the number of previous pregnancies, being longest for the first pregnancies. The duration of labor pain is significantly higher for the respondents who reported the index pregnancy as undesired, and, similarly, the respondents who were reported to be involved with gainful employment would have a shorter duration of labor pain than those having no involvement with gainful employment. PMID- 15799180 TI - The resistance to contraceptive use in young Italian women. AB - OBJECTIVE: To investigate the personal relationship and social factors that influence young women not to use contraceptives. METHODS: A total of 104 young women aged 14-23 years were studied using a 26-item questionnaire which was to be filled out individually. The questionnaire addressed use of contraceptives at the first intercourse and current use, together with various items related to age, educational level, self-image, fantasy about parenthood, and characteristics of the relationships with the partner and the family. A descriptive analysis and a multivariate logistic regression model investigated the relationship between contraceptive non-use and selected variables. RESULTS: Twenty-four percent of the girls did not use any effective contraceptive method at the first sexual intercourse and 21% were current non-users. The study of characteristics associated with inconsistent contraceptive use shows a complex picture where young age, poor knowledge of the partner, an older partner and living in a incomplete family nucleus or outside the family represent significant risk factors. Considering our results as a whole, contraceptive non-use is not only a marker of risk-taking behavior, but sometimes expresses irrational feelings which emerge when facing new sexual experience. PMID- 15799181 TI - A closer look at traditional contraceptive use in Turkey. AB - Withdrawal is the main method used amongst couples in Turkey to prevent pregnancy. Discontinuation of use is most likely to be due to the desire to become pregnant or failure of the method. Withdrawal users are less likely to switch to another contraceptive method; however, among users who do switch, they will most likely switch to a modem method. The strongest three determinants that predict withdrawal use are using withdrawal as one's first method, the woman's age, and the husband's education. The woman's work status and the couple's ethnicity are also important predictors of withdrawal use. Family planning programs should target different segments of the population and focus on correct knowledge of all methods. Results indicate that men and women need to learn about family planning options earlier in their lives or prior to marriage. PMID- 15799182 TI - Factors influencing discontinuation of intrauterine contraceptive devices: an assessment in the Indian context. AB - Sociocultural and behavioral factors are associated with decision to use and the selection of a contraceptive method, continuation of use and reasons for discontinuation of a contraceptive. This paper tries to distinctly outline the determinants of discontinuation of the intrauterine contraceptive device (IUCD), especially in the Indian context. Data on medical reasons for discontinuation are available through clinical trials. However, sociodemographic studies provide one with a wider spectrum to analyze the factors associated with the discontinuation of IUCDs. Information on service providers can be used to improve the quality of family planning services in the country. In India, a thorough review of birth spacing methods, especially the IUCD, is needed since the surveys show a high rate of discontinuation. The emphasis in this paper is upon compilation of reasons for discontinuation of the IUCD and research thereof. The review of the literature is directed towards giving a new direction to assessment of family planning programs, especially in India where population control is largely dependent upon permanent methods. Delineation of factors is important to improve the family planning program. PMID- 15799183 TI - Men and reproductive health in rural Pakistan: the case for increased male participation. AB - OBJECTIVES: In Pakistan, the role of men has always been considered to dominate in the decision-making process pertaining to women's fertility and birth spacing. This study was done to explore men's knowledge, perceptions and behavior on various reproductive health issues. METHODS: A cross-sectional survey was carried out among 180 married adult males in 12 rural districts of Pakistan in 2000. The study was qualitative, utilizing tools such as in-depth and key-informant interviews. RESULTS: The findings pointed out gaps in knowledge and misconceptions among men on a range of reproductive health issues and stress the need for health education. The findings suggest that strategies such as couple counseling, door-to-door campaigns by village-based male family planning workers and small group meetings could be effective. This study indicates a pressing need for incorporating effective intervention strategies, both at the community and the clinic level, backed with efficient counseling, motivation, and provision of services with appropriate education of males in rural areas. CONCLUSION: The findings suggest that investment into promotion of healthy fertility control practices among men will be effective in the near future, as they are interested in acquiring new knowledge and are willing to become partners. PMID- 15799184 TI - The Coraliance study: non-compliant behavior. Results after a 6-month follow-up of patients on oral contraceptives. AB - OBJECTIVES: This follow-up study was planned to establish the frequency with which women miss their contraceptive pill, and to observe their behavior when they forget it. In those women who changed from a continuous cycle to an interrupted type of cycle, or vice versa, the study also aimed to evaluate the impact of this change on the pattern of omission of pills. METHODS: The longitudinal, prospective cohort study included healthy women of child-bearing age for whom a change of pill was being prescribed by their gynecologist. Data were recorded during the 6 months preceding inclusion in the study, and for the 6 months of follow-up; the women were asked to complete a diary in which they recorded the number and exact times of pill omission, and their behavior at each omission. RESULTS: A total of 617 gynecologists included 3316 women into the study; of these, a group of 2418 (73%) revisited the same gynecologist at follow up. The groups who either visited the same or a different gynecologist were similar with respect to age, oral contraception type, omission type and frequency. A large non-compliance rate and women's difficulties in maintaining safe contraception after missing a pill were observed in the group with follow up. Women were never risk-free when they missed a pill; they turned to numerous sources for discordant or conflicting information; 15% of 'not-forgetting' women at the pre-inclusion cycle recorded at least one omission at the last cycle of the 6-month follow-up period. Omission fluctuations during the observational period make it difficult to designate 'forgetful' or 'non-forgetful' classes of women. Administration of the pill in a continuous cycle, and probably 'study' and 'auto-questionnaire' effects, contributed to an improvement in compliance. In the group taking the continuous cycle pill, the omission number slightly decreased, particularly on the first day and week of the cycle, irrespective of the initial cycle type. CONCLUSIONS: The importance of the phenomenon of non-compliance rate is confirmed as well as women's difficulties in knowing how to maintain contraceptive safety. The continuous cycle regimen is likely to improve women's compliance during the critical period of the cycle. PMID- 15799185 TI - Acceptability and side-effects of Implanon in Switzerland: a retrospective study by the Implanon Swiss Study Group. AB - DESIGN: A multicenter, retrospective study of the single-rod contraceptive implant, Implanon (NV Organon, Oss, The Netherlands), was carried out in Switzerland in 1183 women. METHODS: Assessments included duration of use, bleeding pattern, side-effects and subjective acceptability and satisfaction with the method. RESULTS: A total of 991 women (84%) had at least one follow-up visit and 306 (26%) had two visits with a mean duration between insertion and follow-up of 224 days (7.4 months) and 347 days (11.4 months), respectively. Implanon was removed prematurely in 235 women (24%), primarily because of side-effects (20%) and for family planning reasons (4%). Side-effects leading to discontinuation were mainly bleeding disturbances (45%), acne (12%) and other reasons (15%). The mean duration between insertion and removal for discontinuers was 280 days (9.2 months). Side-effects related to bleeding (visit 1) included infrequent bleeding (28%), amenorrhea (33%), prolonged bleeding (15%), and metromenorrhagia (frequent and heavy bleeding) (16%). Other reported side-effects at visit 1 included dizziness (12%), acne (11%), mood swings (8%) and headache (5%). The incidence of side-effects reported at visit 2 was generally comparable to that at visit 1. CONCLUSIONS: The results of this study show that early discontinuation of implant use is primarily due to bleeding problems. Extensive counseling before implant insertion may help to prevent this. It is also important to develop an easy solution to successfully treating progestogen-induced bleeding disturbances. PMID- 15799186 TI - Minimizing missed opportunities: an approach to decrease the unmet need for family planning. AB - OBJECTIVES: The aim of the study was to determine the participants with unmet need and to show that family planning consultancy given to women and men attending the first-level health facilities would increase contraceptive usage and reduce unmet need for family planning. METHODS: All married women aged 15-49 years and men having wives of the same age group attending the health centers for any service other than family planning were included in the study. Among 2021 attendants, a total of 1701 participated in the study (84.2%). Participants were interviewed after they took the services for which they attended. At the end of the inquiries, all willing participants were referred to the family planning unit in the same building. RESULTS: The unmet need (need for any or more effective contraceptive methods) was 43.1%. All participants with unmet need were referred to the family planning unit in the same building and willing participants were given family planning services. Of the participants, 23.9% attended the family planning unit and 17.8% took services immediately at the time of the attendance. CONCLUSIONS: Linking family planning services with other health services can be a cheap and practical way to reduce the unmet need. PMID- 15799187 TI - Acute rehabilitation after severe head injury. PMID- 15799188 TI - Is there a clinical correlate to the histological and radiological evidence of inflammation in trans-ligamentous extruded and sequestered lumbar disc herniaton? AB - The pain of lumbar disc herniation is related to direct compression of the nerve root and to the foreign-body inflammatory response to the herniated disc material. In this study, we attempted to identify disc-related inflammation in trans-ligamentous extruded and sequestered lumbar disc herniation using gadolinium-DPTA-enhanced T1-weighted and short-time inversion recovery (STIR) magnetic resonance imaging. We assessed how these results related to each other, and to patients' clinical status, and to immunohistochemistry findings in disc specimens removed at surgery. Forty-one patients with trans-ligamentous extruded or sequestered disc herniation were investigated clinically and radiologically. Twenty-five patients who did not respond to initial medical treatment or who had progressive neurological deficits underwent discectomy. Specimens of disc material removed during surgery were studied immunohistochemically and inflammatory cell types were identified. The other 16 patients showed good response to treatment, and were given further medical therapy and monitored for a mean of 6.3 months of follow-up. All radiological diagnoses in the operated cases were confirmed at surgery. There were no significant differences among the rates of detection of inflammation by contrast MRI, STIR technique, and immunohistochemistry in the surgical treated cases. In the medically treated cases, the frequency of radiological evidence of inflammation on the initial contrast enhanced T1-weighted and STIR images was similar to the frequency observed on the most recent scans done during follow-up. The study showed that the radiological and immunohistochemical evidence of inflammation in these cases does not correspond with patients' clinical pain profiles over time. It also revealed that STIR and contrast T1W are equally reliable for detecting inflammation in trans-ligamentous extruded or sequestered lumbar disc herniation. In future, studies that focus on physical and chemical mechanisms of pain in lumbar disc herniation in larger series of conservatively and surgically treated patients may clarify the link between inflammation and radicular pain in these types of disc herniation. PMID- 15799189 TI - Uninstrumented ventral surgery for subaxial cervical spine tuberculosis: clinical and radiological outcome. AB - The clinical and radiological outcomes in 14 patients with subaxial cervical spine tuberculosis following uninstrumented anterior decompression surgery and medical treatment were retrospectively reviewed. All the patients underwent an anterior decompression with bone graft followed by immediate mobilization or a period of bed rest for 4 to 6 weeks. The clinical status and whole spine curvature of the cervical spine were assessed preoperatively and at follow up. There was an improvement in the Nurick's grade from a preoperative mean of 2.4 (range 0 - 5) to 1.2 (range 0 - 4) at follow up (p = 0.004). The whole spine curvature showed an improvement in 5 patients, was maintained in 6 patients and showed a kyphotic change in 1 of the 12 patients at follow up. There was evidence of good bony fusion in 12 of the 14 patients for whom data were available at follow up. Anterior decompression with autologous iliac bone graft led to a good clinical and radiological outcome in patients with subaxial cervical spine tuberculosis. PMID- 15799190 TI - Complications of fixation to the occiput-anatomical and design implications. AB - Internal fixation provides an increased immediate stability for the craniovertebral junction; however, there is no current consensus on the optimal method of occipitocervical (OC) fusion. In this report, we present 25 cases of craniovertebral instability treated with OC fusion by plates and screws instrumentation. The 25 cases comprised 12 men and 13 women, whose ages ranged 20 to 78 years. The principal aetiologies that lead to the OC instability of the patients in this series included trauma, rheumatoid arthritis, neoplasm and congenital abnormality. The fusion levels ranged from occiput-C3 to occiput-C6. Two mortalities occurred. The other patients showed satisfactory union after a follow-up of eight to 24 months. OC fusion using a plates and screws system is a safe and highly effective method for providing immediate internal stability to the OC junction. The internal occipital anatomy, which cannot be seen at surgery, is important when dealing with this taxing and potentially dangerous aspect of surgery. PMID- 15799192 TI - Elderly patients with severe head injury in coma from the outset--has anything changed? AB - Advancing age is known to be a determinant of outcome in head injury. We have sought to discover whether there has been any change in the outcome of elderly patients with severe head injury in Newcastle, where these patients have continued to be treated with maximum intervention. A review of prospectively collected data from the Newcastle Head Injury Database for the period 1990 to 2000 was carried out. All patients aged 70 years and above who had sustained a severe head injury (Glasgow Coma Score of 8 or less from the outset) were included. The Glasgow Outcome Score (GOS) was determined at 6 months. Seventy-one patients were identified. Fifty-seven (80%) died and 2 (3%) were in a vegetative state, 11 (16%) had severe disability, 1 (1%) had moderate disability and no patients made a good recovery. The natural history of this condition remains unchanged and due consideration should be given to this when evaluating interventions for elderly patients with a severe head injury. PMID- 15799191 TI - Early prediction of infection after craniotomy for brain tumours. AB - Reliably predicting infection soon after craniotomies would prevent infection and reduce treatment costs and hospitalization expenses. Therefore we analysed potential risk factors and blood count data after craniotomies for brain tumours in order to predict infection as soon as possible after surgery. We analysed 139 patients who underwent craniotomies for brain tumours from January 1997 to December 2001, and divided them into four categories (Types A to D) according to the following: increase in their white blood counts (WBCs) from Day 0 to Day 1, maximum WBC between Day 0 and Day 2, and maximum c-reactive protein (CRP) between Day 0 and Day 4 after surgery. We evaluated potential risk factors and the blood count data for infections via logistic regression analysis. Type D patients had a significantly higher rate of infection (p = 0.0123) than the other Types, while Type B patients had the lowest rate among the four groups (p = 0.0006). When Type A patients suffered CSF leakages, they had a significantly higher possibility of meningitis (p < 0.0001) or scalp infection (p = 0.012). In those Type A or D patients who were male, more than 70 years old, and suffered from metastases from primary lung cancer lesions, the possibility of pneumonia was significantly higher (p = 0.0178). In conclusion, we are able to predict infection within four days after craniotomies for brain tumours according to standard blood count data and certain risk factors. This possibility allows for improved care and better clinical outcomes in patients that undergo craniotomies for brain tumours. PMID- 15799193 TI - Surgical management of ruptured and unruptured symptomatic posterior inferior cerebellar artery aneurysms. AB - This retrospective study analyses the outcome of posterior inferior cerebellar artery aneurysms treated surgically. Thirteen consecutive ruptured and unruptured PICA aneurysm patients from January 1998 to January 2004 were reviewed retrospectively. The mean age was 49.1 +/- 7.4 years. Three were unruptured aneurysms and ten presented with acute subarachnoid haemorrahge. Surgery was performed immediately after completed 4 vessel angiograms using the far lateral approach. Eight were fusiform while five were saccular aneurysms. The saccular aneurysms were clipped. Treatment of the fusiform aneurysms included 3 trappings and three proximal clippings. One patient refused surgery and had endovascular occlusion of a giant PICA aneurysm. Eight out of ten (80%) operated patients needed CSF shunting for hydrocephalus. All check angiograms during follow up demonstrated adequate exclusion of the aneurysms from the circulation. None developed the PICA syndrome clinically or showed infarction on brain scans. All good grade patients recovered without neurological deficits. Our experience showed that early surgery for ruptured PICA aneurysm carries a good prognosis with low morbidity. In aneurysms that cannot be clipped, sacrifice of the PICA without revascularisation procedures in proximally located PICA aneurysms may still be feasible if the occlusion is done distal to the perforators. PMID- 15799194 TI - Pilocytic astrocytoma of the adult--clinical features, radiological features and management. AB - Pilocytic astrocytoma is a histological subtype of astrocytoma classically located in the cerebellum of children and young adults. Cases occurring over the age of 30 years are rare and have not been classified in terms of their clinical features and management. Suitable cases were identified using diagnostic coding and by reviewing a neuropathology database. Casenotes and neuroradiology were reviewed retrospectively. Ten cases were identified over a 6 year period with an incidence of 0.49 cases per million population per year. Tumours were equally distributed between the supra- and infra-tentorial spaces. The most common symptom was headache occurring in 90%. No patient suffered seizures. Total macroscopic treatment remains the treatment of choice. Pilocytic astrocytoma of the adult is a rare tumour with a favourable prognosis. PMID- 15799196 TI - Ventriculo-atrial shunt via the left axillary vein--a novel approach. AB - This report describes a technique to divert cerebrospinal fluid into the right atrium via the left axillary vein in a patient in whom other methods had been attempted without success. PMID- 15799195 TI - High dose radiation induced meningioma. AB - Radiation therapy has played an important integral part in the management of various intracranial rumours. However, radiation has been implicated in the development of intracranial tumours. We describe three cases of radiation-induced meningiomas following high dose cranial irradiation. All these patients developed tumours within the previous radiation field and satisfied the criteria used for the definition of radiation-induced neoplasm. The interval between the irradiation and the onset of meningioma was significantly less in the younger patient. All the cases had several unique features of radiation-induced meningioma including features of atypical meningioma. Two patients developed multiple site meningiomas and one patient developed early recurrence of the tumour. This report confirms that patients exposed to cranial irradiation are at lifelong risk of developing radiation-induced tumours such as meningiomas. Hence these patients require long-term clinical and radiological surveillance to detect occurrence as early as possible. PMID- 15799197 TI - Unusual cervical nerve root arrangement exposed during surgery: case report and review of the literature. AB - Anatomical variations of neural structures in the cervical spine are rare and are not necessarily visible on pre-operative imaging. The authors report an unusual arrangement of neural structures identified during cervical foraminotomy. Anatomical variations of the cervical nerve roots are reviewed and their importance in neurosurgical practice is discussed. PMID- 15799198 TI - Isolated conjugate gaze palsy after frontal lobe tumour surgery. AB - Isolated conjugate gaze palsy following unilateral frontal lobe surgery is uncommon. When it does occur, usually it recovers within hours. We report a case of isolated conjugate gaze palsy which persisted for four weeks post-operatively before recovery commenced. PMID- 15799199 TI - Bilateral testicular enlargement and seminoma in a patient with acromegaly. AB - Growth hormone (GH) excess has been associated with soft tissue changes and an increase in the incidence of colonic polyps and gastrointestinal cancer. We describe a patient with endogenous GH excess caused by a pituitary tumour, resulting in acromegaly. The patient had bilateral testicular enlargement, with tumour of the right testicle. Pituitary MRI revealed a macroadenoma. After resection of the pituitary tumour, GH levels fell below 0.5 ng/ml, with acromegaly resolution. Testicular resection revealed seminoma. Following acromegaly resolution, the enlarged remaining testicle decreased in size. The implications of the testicular enlargement and seminoma in the presence of a GH secreting tumour are discussed. PMID- 15799200 TI - Isolated cervical intramedullary sarcoidosis--a histological surprise. AB - We present a case of cervical intramedullary sarcoidosis. A 56 year old woman presented with progressive paraesthesia affecting the lower limbs. MRI revealed an intramedullar lesion from C4-C7. A laminectomy and subtotal resection was carried out for this presumed intramedullary tumour. Pathology revealed this to be a granulomatous lesion with features indicative of sarcoidosis. Postoperatively, there was no change in her neurological function and her symptoms improved with steroid therapy. PMID- 15799201 TI - Meningioma presenting with transient ischaemic attacks. AB - A case of sphenoid wing meningioma mimicking the presentation of a transient ischaemic attack (TIA) is described. Based on findings from neuroimaging in this case, possible explanations of the mechanism of this uncommon presentation of meningiomas are discussed and the literature is reviewed. PMID- 15799202 TI - Intraradicular lumbar disc herniation: report of two cases and review of the literature. AB - Intraradicular lumbar disc herniation is very rare. The exact mechanism of the dural tear by a herniated disc is not known. Diagnosis of intraradicular lumbar disc herniation is difficult, so that it is rarely suspected preoperatively. This pathological entity may be a factor in the failure of lumbar disc surgery. We present the seventeenth and eighteenth cases of intraradicular lumbar disc herniation. We emphasise its importance and review the literature on intraradicular disc herniation. PMID- 15799203 TI - Misleading imaging features of intracranial dural plasmacytoma: report of two cases. AB - Dural plasmacytomas are very rare and often disregarded in the differential diagnosis of more common lesions of the convexity. We report two cases. Both patients exhibited headache as a main symptom. Computed tomography and magnetic resonance showed a homogeneously enhanced lesion without calcification and without bone involvement. PMID- 15799204 TI - Intracranial plasmacytoma presenting as chronic subdural haematoma. PMID- 15799205 TI - Multiple forms of neurocysticercosis in a single patient. PMID- 15799206 TI - Venous thromboembolism and oral contraceptives: current status and clinical implications. AB - Recent studies of currently available oral contraceptives indicate that the risk of major sequelae is low in young women (aged between 20 and 24 years). Venous thromboembolism remains one event that can occur in users independent of the presence of risk factors. However, the attributable risk is small, with a range of approximately 7 to 18 events per 100 000 women annually. This risk is directly proportional to estrogen dosage starting at levels of 30-35 microg. The type of progestogen (progestin) may also influence risk, though recent studies are controversial. In particular, there is debate surrounding whether desogestrel and gestodene carry a greater risk of thromboembolism than levonorgestrel. Modifiable risk factors for venous thromboembolism include hemostatic disorders, especially factor V Leiden, and possibly obesity. Cigarette smoking is not a significant risk factor in oral contraceptive users. With the exception of avoiding oral contraceptive use among women with a either a personal history of venous thromboembolism or a strong family history (until evaluated for hemostatic abnormalities), and perhaps limiting the use of desogestrel- or gestodene containing oral contraceptives, there is little clinicians can do to reduce the risk of this disorder. PMID- 15799207 TI - Optimizing somatostatin analog therapy in acromegaly: long-acting formulations. AB - Somatostatin peptide analogs have revolutionized the medical treatment of patients with acromegaly. More recent deep intramuscular depot preparations have further improved control, with consistent suppression of growth hormone secretion and optimal lowering of insulin-like growth factor-1. Effective control of growth hormone should, with long-term use, reduce morbidity and mortality from acromegaly and has been shown to result in partial involution of the pituitary adenoma in the majority of treated patients. The currently available depot formulations allow for an injection frequency of 14 days (lanreotide LA 30mg) to 28 days (octreotide LAR 20mg) according to the manufacturers' recommendations. In clinical practice, dose titration by evaluating a growth hormone day profile prior to the next injection can extend the interval between injection (to 6 or even 8 weeks in certain individuals). This is especially true for octreotide LAR, which also has increased flexibility regarding dosage with a 10 and 30mg preparation. The annual 'drug cost' is broadly similar between the two formulations though the additional expenditure on nurse time and clinic visits incurred by an increased injection frequency is a significant consideration. Decreased injection frequency improves acceptability for the patient without a loss in treatment efficacy. A subjective return of typical acromegalic symptoms, such as sweating and headache, also seem to be useful in predicting the timing of the next injection. Other formulations and doses of lanreotide are currently being evaluated, but more interestingly, newer analogs with greater efficacy at the type 5 somatostatin receptor subtype, and pan-receptor analogs, are being developed. These peptides, in conjunction with the likely availability of a growth hormone receptor blocking agent (pegvisomant), will further expand the medical therapy options for patients with acromegaly. PMID- 15799208 TI - Identification and treatment of eating disorders in women with type 1 diabetes mellitus. AB - A series of case reports in the early 1980s and prevalence studies in the 1990s highlighted the serious medical consequences of coexisting eating disorders and diabetes mellitus. Diabetes-specific treatment issues, such as the need to carefully monitor diet, exercise, and blood glucose, may contribute to the development of eating disorder symptoms among women with diabetes mellitus. The attention to food portions and bodyweight that is part of routine diabetes mellitus management parallels the rigid thinking about food and body image found in women with eating disorders who do not have diabetes mellitus. Additionally, intensive insulin management of diabetes mellitus, the current standard of care, has been shown to be associated with bodyweight gain. Following from this, it may be that the very goals of state-of-the-art diabetes mellitus care increase the risk for developing an eating disorder. Once an eating disorder and recurrent insulin omission becomes entrenched, a pattern develops which is hard to break - one of chronic hyperglycemia, depressed mood, fear of bodyweight gain, and frustration with diabetes management. Eating disorders predispose women with diabetes mellitus to many complex medical risks. For example, insulin omission and reduction, eating disorder symptoms unique to diabetes mellitus, are strongly associated with an increased risk of diabetic ketoacidosis and with microvascular complications of diabetes mellitus such as retinopathy. For this reason, it is critical that diabetes mellitus clinicians understand more about eating disorders so as to improve the likelihood of early detection, appropriate treatment, and prevention of acute and long-term medical complications within this high-risk group of women. PMID- 15799209 TI - Treatment of microalbuminuria in patients with type 2 diabetes mellitus. AB - The incidence of type 2 diabetes mellitus is increasing world-wide, and is now one of the leading causes of end-stage renal disease in Western countries. Type 2 diabetes mellitus is also a major risk factor for cardiovascular events. Therefore, the early identification of patients at greatest risk, and the subsequent initiation of renal and cardiovascular protective treatments, are of the utmost importance. Microalbuminuria refers to a subclinical increase in urinary albumin excretion. By definition it corresponds to an albumin excretion rate of 20 to 200 microg/min (30 to 300 mg/day) or an albumin to creatinine ratio (mg/mmol) of 2.5 to 25 in males and 3.5 to 35 in females. Microalbuminuria is an important clinical finding because it is not only associated with an increased risk of progression to overt proteinuria (macroalbuminuria) and renal failure, but also cardiovascular events. In patients who progress to overt nephropathy, microalbuminuria usually precedes macroalbuminuria by an interval of 5 to 10 years. In patients with type 1 diabetes mellitus, blood pressure increases and renal function declines after the onset of macroalbuminuria. However, in patients with type 2 diabetes mellitus, hypertension and a decline in renal function may occur when albumin excretion is still in the microalbuminuric range. Large clinical trials have demonstrated that achieving tight glycemic (i.e. glycosylated hemoglobin < 7.0%) and blood pressure (i.e. < 130/85mm Hg) control retards the progression of renal disease. There is accumulating evidence to suggest that the use of antihypertensive agents which target the renin angiotensin system (RAS) can slow the progression of renal disease and provide cardioprotection in patients with type 2 diabetes mellitus and microalbuminuria. Antihypertensive agents which target the RAS also appear to have advantages over and above reductions in systemic blood pressure. In summary, the annual screening of patients with type 2 diabetes mellitus for microalbuminuria, and the initiation of measures to retard the progression of renal and cardiovascular disease, are now considered part of routine clinical practice. In particular, the finding of microalbuminuria should provoke an intensified modification of the common risk factors for renal and cardiovascular disease, that is hyperglycemia, hypertension, dyslipidemia and smoking. Antihypertensive therapy in patients with microalbuminuria and type 2 diabetes mellitus should be initiated with angiotensin converting enzyme (ACE) inhibitors or angiotensin-II type 1 receptor antagonists. PMID- 15799210 TI - Parathyroid hormone, its fragments and their analogs for the treatment of osteoporosis. AB - The susceptibility to traumatic fracturing of osteopenic bones, and the spontaneous fracturing of osteoporotic bones by normal body movements caused by the microstructural deterioration and loss of bone, are currently treated with antiresorptive drugs, such as the bisphosphonates, calcitonin, estrogens, and selective estrogen receptor modulators. These antiresorptive agents target osteoclasts and, as their name indicates, reduce or stop bone resorption. They cannot directly stimulate bone formation, increase bone mass above normal values in ovariectomized rat models, or improve microstructure. However, there is a family of agents - the parathyroid hormone (PTH) and some of its fragments and their analogs - which directly stimulate bone growth and improve microstructure independently from impairing osteoclasts. These drugs are about to make their clinical debut in treating patients with osteoporosis and, probably not too far in the future, for accelerating fracture healing. They stimulate osteoblast accumulation and bone formation in three ways via signals from the type 1 PTH/PTH related protein (PTHR1) receptors on proliferatively inactive preosteoblasts, osteoblasts, osteocytes and bone-lining cells. The receptor signals shut down the proliferative machinery in preosteoblasts and push their maturation to osteoblasts, cause the osteoblastic cells to make and secrete several factors that stimulate the extensive proliferation of osteoprogenitors without PTHRI receptors, stimulate the reversion of bone-lining cells to osteoblasts, and extend osteoblast lifespan and productivity by preventing them from suicidally initiating apoptosis. The first of the PTHs to reach the clinic will be teriparatide [recombinant human (h)PTH-(1-34)], which was recommended for approval in 2001 by the US Food and Drug Administration Endocrinology and Metabolic Drugs Advisory Committee for the treatment of postmenopausal osteoporosis. Teriparatide has been shown to considerably increase cancellous and cortical bone mass, improve bone microstructure, prevent fractures and thus provide benefits that cannot be provided by current antiresorptive drugs, when administered subcutaneously at a daily dose of 20 microg for no longer than 2 years to patients with osteoporosis. PMID- 15799212 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- 15799211 TI - Spotlight on estradiol-intranasal in the management of menopause. AB - Estradiol-intranasal is a nasal spray formulation containing an aqueous solution of 17beta-estradiol that has a unique pulse-like pharmacokinetic profile. In a well designed, placebo-controlled trial estradiol-intranasal 200 to 400 microg/day significantly reduced the incidence and severity of climacteric symptoms in women with moderate to severe menopausal symptoms after 4 and 12 weeks' treatment. The efficacy of estradiol-intranasal 300 microg/day was similar to that of oral estradiol 2 mg/day in this and another double-blind placebo controlled trial. This equivalent efficacy was maintained in a subgroup of women with initially severe symptoms, and in smokers. Reductions in the incidence of atrophic vaginal mucosa and genitourinary symptoms and increases in the karyopyknotic index achieved with estradiol-intranasal 300 microg/day were also similar to those observed with oral estradiol 2 mg/day. Assessments of the effects of estradiol-intranasal on the complications of menopause (increased risk of cardiovascular disease and osteoporosis) are ongoing; however, estradiol intranasal (sequentially combined with a progestogen) produced significant beneficial effects on some lipid parameters and on markers of bone resorption and formation, and bone mineral density in postmenopausal women. Estradiol-intranasal had no significant effects on serum levels of most of the assessed hemostatic factors, or on angiotensinogen or insulin levels. Estradiol-intranasal 100 to 600 microg/day was generally well tolerated in clinical trials and most adverse events were mild to moderate. The most commonly reported events were nasal symptoms and mastalgia. There was no evidence of endometrial hyperplasia with up to 1 year's treatment with estradiol-intranasal 300 microg/day combined with a progestogen. The incidence of mastalgia and withdrawal or breakthrough bleeding was lower with estradiol-intranasal 300 microg/day than with oral estradiol 2 mg/day (both administered with a progestogen) in one trial. In another trial, the incidence of mastalgia was lower with estradiol-intranasal 300 microg/day than with estradiol transdermal 50microg (both administered with a progestogen). However, the overall incidence of adverse events was similar between the two treatments in this trial. In conclusion, estradiol-intranasal 200 to 400 microg/day (optimal initiating dose 300 microg/day) reduces the incidence and severity of menopausal climacteric symptoms and has a good tolerability profile. Thus, evidence to date suggests that estradiol-intranasal is a useful treatment option for menopausal symptoms. PMID- 15799213 TI - Impaired glucose tolerance: is there a case for pharmacologic intervention? AB - Impaired glucose tolerance (IGT) is determined by measuring plasma glucose levels 2 hours after glucose loading in the oral glucose tolerance test. There is good evidence from epidemiologic and prospective trials [e.g. Diabetes Epidemiology: Collaborative Analysis of Diagnostic Criteria in Europe (DECODE)] linking IGT with the development of type 2 diabetes mellitus and cardiovascular disease (CVD). IGT is characterized by an increase in postprandial glucose levels, which is considered the earliest metabolic abnormality in type 2 diabetes mellitus. It is one of a series of risk factors for CVD (hypertension, high triglyceride levels, low high-density lipoprotein-cholesterol and central obesity), known as the metabolic syndrome. The different factors making up this syndrome are intimately related. An impaired lipid profile can contribute to insulin resistance, as IGT may play a pathogenic role on other cardiovascular risk factors. IGT is the first easily identifiable step in the pathophysiology of type 2 diabetes mellitus. It is associated with high risk for type 2 diabetes mellitus and subsequent vascular morbidity and mortality. It is currently unknown whether treating IGT will reduce the incidence of macrovascular complications, as studies addressing this issue have yet to be conducted. Therefore, the main reason to identify and treat IGT is to prevent or delay the onset of type 2 diabetes mellitus. It has been demonstrated that lifestyle intervention with diet and exercise can reduce the incidence of type 2 diabetes mellitus. Pharmacologic intervention with metformin and acarbose is also effective. Other drugs, such as those indicated to treat other parameters of the metabolic syndrome, may also be useful. We can now be assured that prevention or delay of onset of type 2 diabetes mellitus is possible in individuals with IGT, either by changes in lifestyle or by pharmacotherapy. PMID- 15799214 TI - Subclinical hypothyroidism: how should it be managed? AB - The term 'subclinical hypothyroidism' applies to patients who have mildly increased levels of serum thyrotropin hormone (TSH) and normal levels of thyroxine and liothyronine (triiodiothyronine). This very common condition, also called 'mild thyroid failure', accounts for 75% of patients who have increased serum TSH. For patients with sustained increases above 10 mIU/L, there is uniform agreement that thyroxine therapy is indicated. Therapy for milder forms of hypothyroidism is controversial. Some randomized clinical trials favor therapy for mild thyroid failure, but they are inconclusive because they lack stratification for the subgroup of patients with TSH levels below 10 mIU/L. For this subgroup, we recommend individualized management. The presence of goiter, positive thyroperoxidase (TPO) antibodies, manic-depressive disorder, fertility problems, or pregnancy or the anticipation of pregnancy favors the initiation of therapy. Positive TPO antibodies are a strong indication for therapy because of the high likelihood in these patients of progression to overt hypothyroidism; patients who are already receiving thyroxine should have adjustments of their dosage. Children and adolescents with mild thyroid failure should also be treated because of possible adverse effects on growth and development. It has been suggested that subclinical hypothyroidism is a cardiovascular risk factor, however further investigation is needed. The controversy surrounding therapy will not be resolved until more randomized studies are available for the subgroup of patients with TSH <10 mIU/L, and until the question of cardiovascular risk factors is further clarified. PMID- 15799215 TI - Male hormonal contraception: an update on research progress. AB - New male contraceptive options are needed to provide couples with additional well tolerated, effective, and reversible contraceptive choices. Research and development of new male hormonal contraceptives have produced many exciting possibilities over the past 20 years. Two large, multicenter, World Health Organization (WHO) trials demonstrated that high-dosage testosterone provides effective suppression of follicle-stimulating hormone, luteinizing hormone, and spermatogenesis. In men whose sperm counts suppress to < or = 3 million/ml, high dosage testosterone provides contraceptive efficacy similar to female oral contraceptives. Since the WHO efficacy studies, investigators have focused on evaluating combination regimens that include lower dosages of testosterone plus gonadotropin-releasing hormone (GnRH) analogs or progestogens (progestins). In small, randomized trials, combination regimens of testosterone plus GnRH analogs or progestogens have produced more uniform suppression of spermatogenesis than high dosage testosterone alone. The major adverse effects of most of the studied male hormonal contraceptives are modest bodyweight gain and suppression of high density lipoprotein-cholesterol levels. The latest research studies indicate that a well-tolerated, effective male hormonal contraceptive may soon be developed. PMID- 15799216 TI - Somatropin and glucose homeostasis: considerations for patient management. AB - More than 60 years ago it was shown, in dogs, that anterior pituitary extracts may cause glucose intolerance and that hypophysectomy was associated with increased insulin sensitivity. Accordingly, active acromegaly is characterized by insulin resistance at the hepatic and muscular level, whereas children with growth hormone (GH) deficiency are insulin hypersensitive and prone to developing fasting hypoglycemia. Somewhat unexpectedly, hypopituitary adults with untreated GH deficiency tend to be insulin resistant, which may be aggravated by somatropin (GH) therapy. The explanation for this apparent paradox has not been fully established. It is, however, likely that high circulating levels of free fatty acids (FFA) are responsible for insulin resistance, both before and after somatropin therapy. In the untreated state, patients have abdominal obesity, which increases circulating FFA levels. Since GH has potent lipolytic effects, somatropin therapy will further increase FFA levels. Theoretically, this GH replacement effect will eventually be compensated for by favorable alterations in body composition, including a reduction of fat mass. Subcutaneous somatropin therapy, however, will cause some degree of hypersomatropinemia in the prandial phase, which will inevitably antagonize the physiologic effects of insulin. At present, the best way to circumvent this inherent problem is to employ evening injections of somatropin and to ensure that the dosage is not too high. In the latter regard, it is important to realize that dosage requirements are lower in adults compared with children, and that the dosage will probably need to be reduced with age in the individual patient. PMID- 15799217 TI - Optimizing insulin therapy in pregnant women with type 1 diabetes mellitus. AB - Pregnancy complicated by type 1 diabetes mellitus is associated with an increased risk of complications in the mother and infant. Normal or near normal glycemic control prior to and during pregnancy reduces many of these risks to levels observed in the general population. This degree of glycemic control is generally achievable only with intensive insulin therapy: multiple daily injections (MDI) or continuous subcutaneous insulin infusion (CSII) via an insulin pump. These therapeutic regimens have been found to result in comparable glycemic control, although CSII provides increased flexibility in terms of patient lifestyle, and may reduce the incidence of hypoglycemia. Frequent home blood glucose monitoring is imperative during pregnancy in order to optimize glycemic control and reduce the risk of hypoglycemia. Furthermore, insulin requirements change significantly over the course of pregnancy. The new short-acting insulin analogs, insulin lispro and insulin aspart, have pharmacodynamic properties which make them ideal for use during pregnancy. Although the number of published studies evaluating the use of insulin lispro during pregnancy is limited, the majority support its safety. No studies of insulin aspart in pregnancy have been published in full. In addition to optimization of glycemic control, frequent assessment for development and/or progression of microvascular complications is necessary during pregnancy. PMID- 15799218 TI - Modern therapy for Paget's disease of bone: focus on bisphosphonates. AB - There has been a dramatic change in the therapeutic approach to patients with Paget's disease of bone over the last 40 years. In the 1960s, only symptomatic therapy could be given, with control of pain the main objective. Analgesics and nonsteroidal anti-inflammatory drugs were the most commonly used agents. From 1968 onwards, antiosteoclastic agents became available, including calcitonin, plicamycin (mithramycin) and etidronate (etidronic acid), a first-generation bisphosphonate. Limitations with these agents, including potentially deleterious effects on bone mineralization with etidronate (etidronic acid), has cleared the way for increasingly potent second- and third-generation bisphosphonates, including clodronate (clodronic acid), pamidronate (pamidronic acid), alendronate (alendronic acid) and risedronate (risedronic acid). Even more potent bisphosphonates will become available in the near future. With the newer bisphosphonates, there is some hope for long-term remission (if not definitive healing) of pagetic lesions, as well as prevention of long-term complications in both symptomatic and asymptomatic patients. Thus, indications for therapy have been extended to include younger patients to prevent bone deformity of the limbs and skull, leading to secondary osteoarthritis, facial deformities and potentially to sarcoma transformation; as well as to elderly patients to prevent bone fragility, leading to fracture, and pagetic vascular steal syndromes. The increased potency and longer duration of action of newer bisphosphonates more than compensates for their marginally increased cost compared with older bisphosphonates. PMID- 15799220 TI - Hydrophilized poly(lactide-co-glycolide) nanospheres with poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) triblock copolymer. AB - A novel method for preparing the PLGA nanospheres with hydrophilic surface has been designed and characterized. Because of good solubility of tetraglycol in water, PLGA (poly(lactide-co-glycolide)) nanospheres were formed by spraying the PLGA/tetraglycol solution into water. The size of PLGA nanospheres was manipulated by changing the concentration of PLGA/tetraglycol solution. Based on the hydrophobic interaction between PLGA and poly(propylene oxide) domain of F 127 (one of Pluronics, poly(ethylene oxide)-poly(propylene oxide)poly(ethylene oxide) triblock copolymer, F-127-coated PLGA nanospheres was prepared to enhance the stability of PLGA nanospheres in the aqueous media. For the application as a drug delivery vehicle, it was characterized by measuring the loading amount, the encapsulation efficiency and the release pattern of drug. Paclitaxel used as a potent anti-cancer drug was selected as a model drug. PMID- 15799221 TI - Formulation and evaluation of novel tableted chitosan microparticles for the controlled release of clozapine. AB - Controlled release formulations of clozapine microparticulated tablets were prepared by using chitosan. Microparticles were characterized for particle size and size distribution. Microparticles were compressed into tablets using the directly compressible excipients. SEM photographs of the fractured part of the tablet revealed the presence of discrete particles in the tablets, suggesting that the system chosen is ideal for tableting. Drug release from the tableted microparticles exhibited an initial burst effect, but the release decreased with increasing extent of cross-linking. Tablets were coated with chitosan or cellulose acetate, which significantly lowered the initial burst effect when compared to uncoated tablets. Drug release from chitosan-coated tablets was slightly higher than the tablets coated with cellulose acetate. Tablets prepared were effective in delivering clozapine over a period of 12 h. PMID- 15799222 TI - In vivo and in vitro characteristics for insulin-loaded PLA microparticles prepared by w/o/w solvent evaporation method with electrolytes in the continuous phase. AB - Insulin-loaded poly(lactide) (PLA) microparticles were successfully prepared by 6% w/v PLA in the organic phase, 10% w/v PVP and varied types of 5%w/v electrolytes in the continuous phase, by using a water-in-oil-in-water emulsion/ solvent extraction technique. Addition of electrolytes such as NaCl, CaCl2 into the external phase significantly improved insulin entrapment efficiency compared to the case of no additives. NaCl was the most effective for obtaining high entrapment efficiency, with microparticle yield 81.2%, trapping efficiencies 49%, insulin-loading level 5.5% w/w and mean particle size 14.8 microm. The distribution (%) of insulin on the PLA microparticles surface, outer layer and core were 8, 37 and 43%, respectively. The cumulative release of insulin had an upper limit of approximately 24% of the insulin load at 24 days. A steady release rate was 0.5 microg insulin/mg microparticles/day of insulin release maintained for 24 days. Total protein-leaking amount was reduced after addition of electrolytes in the continuous aqueous phase. Rabbit glucose levels were evaluated after subcutaneous 20 mg insulin-loaded PLA microparticles or PLA blank microparticles. Study results show that the insulin-loaded PLA microparticles significantly reduced the glucose level than PLA blank microparticles. The insulin-loaded PLA microparticles, physicochemical characterization data and the animal result obtained in this study may be relevant in optimizing the PLA microparticle formulation incorporation and delivery insulin carriers. PMID- 15799223 TI - Development and characterization of different low methoxy pectin microcapsules by an emulsion-interface reaction technique. AB - In the controlled release area, biodegradable microcapsules are one of the most useful devices to deliver materials in an effective, prolonged and safe manner. A new charged film microcapsular carrier system, using three different pectins, is described. The study utilized pectin microcapsules prepared by two encapsulation mechanisms of interfacial reaction explored through interaction of charged droplet-oil-anionic surfactant-calcium or oil-cationic surfactant with negatively charged pectin. A method for drug encapsulation was developed based on the type of pectin, surfactants and emulsification technique. Both types of surfactant, anionic sodium dodecyl sulphate (SDS) and cationic benzalkonium chloride (BzACl) promoted polymer film formation on the oil droplet surfaces, probably through cross-linking and electrostatic interaction, respectively. Microcapsules consisting of pectin as shell and hydrophobic oil as core were characterized. The resulting microcapsules were relatively small particles (d< 3 microm), had high total particle number, specific surface area and drug encapsulation efficiency. They also demonstrated good stability with minimum particle aggregation. Correlation between physicochemical and drug release kinetic parameters were investigated with regard to the effect of pectin macromolecular structure and nature of surfactant used as a counterion in the manufacturing of microcapsules. The release rate of the encapsulated material (prednisolone) in three microcapsules can be controlled by manipulating the conformational flexibility of pectins in the presence of different counterions. As a result, biodegradable pectin microcapsules offer a novel approach for developing sustained release drug delivery systems that have potential for colonic drug delivery. PMID- 15799224 TI - Biodegradable microspheres of ketorolac tromethamine for parenteral administration. AB - Ketorolac tromethamine loaded microspheres were prepared using two different polyesters, namely poly (lactic acid) and poly (glycolic acid) by solvent evaporation technique. The morphology of microspheres was analysed by scanning electron microscopy. In vitro release profiles of these microspheres were studied in phosphate buffered saline pH 7.4. The release kinetics of ketorolac tromethamine from the microspheres was evaluated by fitting the release data to the zero-order, Higuchi and korsemeyer-peppas equations. All microspheres showed initial burst release, followed by fickian diffusion of drug through microspheres. These microspheres were formulated as parenterals to have controlled release system. PMID- 15799225 TI - In vitro cytotoxicity of mitoxantrone-incorporated albumin microspheres on acute promyelocytic leukaemia cells. AB - In the present study, the preparation and characterization of bovine serum albumin (BSA) microspheres and the evaluation of the in vitro cytotoxicity of these microspheres on acute promyelocytic leukaemia (HL-60) cells were described. Mitoxantrone (MTZ)-incorporated microspheres were evaluated for particle size, drug loading, release characteristics and surface morphology. The biological effect of MTZ released from BSA microspheres was determined on an in vitro cultured HL-60 cell line, showing that, after encapsulation, MTZ still retains cytotoxic activity. For this purpose, methyl-thiazol-tetrazolium (MTT) assay was used to evaluate the in vitro cytotoxicity of MTZ-loaded microspheres. Particle size of BSA microspheres was determined between 17.61-20.38 microm and they were smooth and spherical in shape. Encapsulation efficiency of the drug-loaded microspheres was between 22.26-60.50%. For MTZ-containing microspheres, the cell death ratios were greater than 80% for all formulations. This study demonstrate that BSA microspheres were well suited for the controlled release of MTZ and were promising for anti-cancer chemotherapy. PMID- 15799226 TI - Nasal delivery of insulin using chitosan microspheres. AB - Nasal delivery of insulin is an alternative route for administration of this drug. The objective of this study was preparation of chitosan microspheres for insulin nasal delivery. After preparation of insulin chitosan microspheres by emulsification-cross linking process, the effect of chitosan quantity (200 400mg), cross-linker type (ascorbic acid or ascorbyl palmitate) and amount (70 140 mg) were studied on the morphology, particle size, loading efficiency, flow and release of insulin from the microspheres by a factorial design. Optimized formulation was administered nasally in four groups of diabetic rats and their serum insulin levels were analysed by the insulin enzyme immunoassay kit and the serum glucose by the glucose oxidase kits. Insulin loading in microspheres was between 4.7-6.4% w/w, preparation efficiency more than 65% and mean particle size was 20-45 microm. In most cases, drug released followed a Higuchi model. Ascorbic acid caused an increase in stability, particle size and T50%, while decreased the loading efficiency and production efficiency. Increasing the chitosan content, increased particle size, flow and insulin release rate form the microspheres. The increase of cross-linking percentage decreased the flow and size of the microspheres while increase of cross-linking percentage promoted the stability and decreased DE8% of insulin. Microspheres containing 400mg of chitosan and 70mg ascorbyl palmitate caused a 67% reduction of blood glucose compared to i.v. route and absolute bioavaliability of insulin was 44%. The results showed that chitosan microspheres of insulin are absorbable from nasal route. PMID- 15799227 TI - An improvement of double emulsion technique for preparing bovine serum albumin loaded PLGA microspheres. AB - A modified double emulsion technique was adopted to prepare bovine serum albumin (BSA) loaded poly (D,L-lactic-co-glycolic acid) (PLGA) microspheres. In the formulations, polysorbates (Tween) such as Tween20, Tween40 or Tween80, instead of frequently used poly (vinyl alcohol) (PVA), was used as the emulsifier. Microspheres with porous surface, large particle size, low microsphere yield (approximately 65.4%) and BSA entrapment efficiency (approximately 25.2%) were obtained when Tween80 aqueous solution alone was used as the outer aqueous phase. However, microspheres with smooth surface, high yield and BSA entrapment efficiency could be produced successfully by introducing sodium chloride or glucose into the outer aqueous phase. Adding 5.0%(w/v) sodium chloride into the continuous phase led to increase in microsphere yield and BSA entrapment efficiency from 65.4% and 25.2% to approximately 100% and 76.6%, respectively. Microsphere yield and BSA entrapment efficiency increased from 64.5% and 25.2% to 97.2% and 89.3%, respectively, when 15.0%,(w/v) glucose was added into the continuous phase. In constrast to the microspheres prepared in the presence of additive, a more marked burst release was observed for microspheres prepared without additive in the continuous phase, which may be attributed to the porous morphology of the latter. PMID- 15799228 TI - On the diameter and size distributions of Bovine Serum Albumin (BSA)-based microspheres. AB - A fluid mechanics-based correlation for the average size of Bovine Serum Albumin (BSA) microspheres, prepared using a water-in-oil emulsion technique, is presented. The correlation is formulated based on the theory of turbulent dispersion and a non-dimensional Weber number. Measured average diameters of the BSA microspheres prepared in olive oil at different stirring speeds are used to construct the correlation formula. The correlation gives good agreement with experimentally measured average diameters for a wide range of Weber numbers. This correlation is particularly useful to the pharmaceutical industry for predicting the size of encapsulated microspheres used in drug delivery prior to microsphere preparation. The effect of additives on microsphere size was also explored. The average diameter of the BSA microspheres was doubled by the addition of a bioadherent vitelline protein B solution. In addition, a Rosin-Rammler statistical distribution function is shown to accurately represent the microsphere size distribution obtained at different stirring speeds. PMID- 15799229 TI - Literature alerts. PMID- 15799230 TI - Secular change in height, sitting height and leg length in rural Oaxaca, southern Mexico: 1968-2000. AB - OBJECTIVE: To evaluate secular changes in height, sitting height and estimated leg length between 1968 and 2000 in residents in a rural Zapotec-speaking community in Oaxaca, southern Mexico. MATERIALS AND METHODS: Height and sitting height were measured in school children 6-13 years (1968; 1978, 2000), in adolescents 13-17 years (1978, 2002) and adults 19-29 years (1978, 2000). Leg length was estimated as height minus sitting height. The sitting height/ height ratio was calculated. Subjects were grouped by sex into four age categories: 6-9, 10-13, 13-17 and 19-29 years for analysis. The Preece-Baines Model I growth curve was fitted to cross-sectional means for 1978 and 2000. RESULTS: There were no differences between children 6-9 and 10-13 years in 1968 and 1978 with the exception of the sitting height ratio in girls 6-9 years. Children of both sexes 6-13 years and adolescent boys 13-17 years were significantly larger in the three dimensions in 2000 compared to 1978; adolescent girls differed only in height and sitting height. Adult males in 2000 were significantly taller with longer legs than those in 1978, but the samples did not differ in sitting height and the ratio. Adult females in 1978 and 2000 did not differ significantly in the three dimensions. Rates of secular change in height and sitting height between 1978 and 2000 were reasonably similar in the three age groups of male children and adolescents, but the rate for estimated leg length was highest in 10-13-year-old boys. Secular gains were smaller in adult males, but were proportionally greater in estimated leg length. Girls 6-9 and 10-13 years experienced greater secular gains in height, sitting height and estimated leg length than adolescent and young adult females, while secular gains and rates decreased from adolescent girls to young adult women. Ages of peak velocity for height, sitting height and estimated leg length declined in boys, while only ages of peak velocity for height and estimated leg length declined in girls. CONCLUSIONS: There are major secular increases in height, sitting height and estimated leg length of children and adolescents of both sexes since 1978. Secular gains in height are of similar magnitude in boys and girls 6-13 years, but are greater in adolescent and young adult males than females. The secular increase in height of young adults of both sexes is smaller than that among adolescents. Estimated leg length accounts for about 60% of the secular increase in height in children of both sexes. Estimated leg length and sitting height contribute equally to the secular increase in height in adolescent boys, whereas estimated leg length accounts for about 70% of the secular increase in height in young adult males. Sitting height contributes about two-thirds of the secular increase in height in adolescent and young adult females. PMID- 15799231 TI - Secular change in age at menarche in rural Oaxaca, southern Mexico: 1968--2000. AB - OBJECTIVE: To evaluate secular change in the age at menarche between 1978 and 2000 in residents of a rural Zapotec-speaking community in Oaxaca, southern Mexico, using status quo and retrospective methods. MATERIALS AND METHODS: Status quo menarcheal status of girls 9-18 years of age in 1978 (n= 101) and 2000 (n=238) and retrospective ages at menarche of adult women 19+ years of age in 1978 (n = 228) and 2000 (n = 246) were obtained via interview. Probit analysis was used to estimate median ages at menarche and 95% confidence intervals (CI) for the status quo data. Analysis of variance and linear regression was used to compare the retrospective data across surveys. RESULTS: The median ages at menarche of adolescents are 14.8+/-1.2 years (0.24 year, 95% CI 14.2-15.4 years) in 1978 and 13.0+/-1.0 years (0.10 year, 95% CI 12.7-13.3 years) in 2000. Age at menarche has declined by 1.8 years over about 23 years, 0.78 year/decade (95% CI = 0.65-0.91 year/decade). Recalled ages at menarche do not differ by age group in the 1978 survey, but differ significantly by age group in the 2000 survey (p <0.001). Within the 2000 survey, the two youngest age groups (< 29, 30-39 years) do not differ, but attain menarche earlier than women in the four older age groups (p < 0.05), who do not differ from each other in age at menarche. The estimated rates of secular decline in age at menarche in adult women vary between 0.38 and 0.42 years/decade (0.26-0.56 year/decade). CONCLUSIONS: There is a major secular decline in the age at menarche of adolescent girls and young adult women between 1978 and 2000. The estimated rate of decline in adult women is about one half of that in adolescent girls. The secular decline in age at menarche is consistent with corresponding secular gains height, sitting height and estimated leg length of children and adolescents in the community; corresponding secular gains are smaller in young adults 19-29 years. PMID- 15799232 TI - A cross-sectional examination of growth indicators from Nicaraguan adolescent girls: a comparison of anthropometric data from their Guatemalan counterparts. AB - BACKGROUND: Few studies have examined growth data from adolescent girls in Latin America and almost none have been conducted in Nicaragua. Thus this study examines growth data from Nicaraguan adolescent girls. AIM: This study describes cross-sectional growth data from a sample of Nicaraguan girls. These data are compared with data from Mexican-US and Guatemalan girls. It is hypothesized that (a) Mexican-US and Guatemalan wealthy ladina girls will show better indicators of nutritional status when compared with the Nicaraguan girls, and (b) the Nicaraguan girls will show similar anthropometric indicators when compared with poor Guatemalan girls. SUBJECTS AND METHODS: Anthropometric and age at menarche data were collected from a sample of 154 adolescent girls ages 10-17 years living in Managua, Nicaragua. RESULTS: The findings revealed that these girls are significantly shorter and lighter than US, Mexican-US and wealthy Guatemalan adolescent girls. These girls are also significantly taller and heavier than their poor Guatemalan counterparts. CONCLUSION: This study reveals that the Nicaraguan girls are undernourished when compared with other Latin American girls living in an improved overall environment. The data also reveal that the Nicaraguan girls have better indicators of nutritional status when compared with a cohort of poor Guatemalan girls. PMID- 15799233 TI - Genetic variation and relationships at six VNTR loci in two distinct sample populations in Brazil. AB - BACKGROUND: The Brazilian population has been the focus of intensive genetic study due to admixture characteristics whereas there are few reports on the variability of VNTR loci in Brazil. PRIMARY OBJECTIVE: The aim of this study was to analyse genetic parameters in sample populations from two geographically distant regions: Sao Luis City, in Maranhao State and Campinas City, in Sao Paulo State. We investigated if distinct colonization influences could produce detectable differences in genetic background. SUBJECT AND METHODS: DNA samples from peripheral drained blood were obtained from unrelated individuals who underwent paternity testing. Allelic variation in six VNTR loci (D2S44, D4S139, D5S110, D8S358, DI0S28 and D17S79) was evaluated. The results were compared to reference databases available for general Latin-derived European and African American populations as well as for other Brazilian groups. RESULTS: This study reveals that forensic population parameters did not show differences among regions, although we detected admixture values varying between the south-east and north-east of Brazil. CONCLUSIONS: Differences between the two samples are probably due to different admixture proportions of European- and African-derived alleles in each region: both populations are in Hardy Weinberg equilibrium. In addition, the allelic frequency for all loci, in both populations, can be used as database for forensic purposes. PMID- 15799234 TI - Admixture estimates for Churuguara, a Venezuelan town in the State of Falcon. AB - BACKGROUND: The present Venezuelan population is the admixture product of Amerindians, Europeans and Africans, a process which was not homogeneous over the country. Blood groups, STRs and VNTRs, specifically D1S80, have been used successfully in admixture studies, but few have been made in Venezuela. AIM: This study aims to estimate the admixture components of Churuguara, Venezuela, and to evaluate the genetic relationship of this population with other Venezuelan as well as worldwide populations through principal component analysis and the study of dendrograms based on genetic distances. SUBJECTS AND METHODS: Gene frequencies of blood groups ABO and Rh (only anti D), of STRs VWA, F13A01, FES/FPS and VNTR D1S80 were studied in a sample of 60 individuals born in Churuguara, a Venezuelan town of admixed ancestry in the State of Falc6n. Admixture was estimated with Chakraborty's gene identity method, and Nei's standard genetic distance was used to build two dendrograms with the neighbour-joining approach, one based on the three STRs and the other based only on D1S80. Principal component analyses with the gene frequencies of these markers were also performed. RESULTS: The frequency of allele ABO*O was 0.788, of ABO*A was 0.187 and of RH*D was 0.74. D1S80 showed 16 different alleles with a heterozygosity of 0.880, whilst the three STRs showed only eight different alleles and heterozygosities between 0.733 and 0.797. The estimates of admixture obtained in this analysis were 52.5% for the Spanish parental group, 27.6% for the African and 19.9% for the Amerindian. Comparison of Churuguara with other Latin American populations shows that its African component is not as high as that observed in Colombian Choco, but it is higher than that observed in other samples from Colombia, Chile and Maracaibo (Venezuela). CONCLUSIONS: Results of the admixture analysis are consistent with those obtained with two dendrograms and principal component analyses, suggesting that the strong initial Amerindian component of 500 years ago has been diluted by the continuous flow of European genes, mainly Spanish, to this region. PMID- 15799235 TI - Reference charts of body proportion for Japanese girls and boys. AB - BACKGROUND: Data from previous studies provided the mean values of body proportions for each age group of healthy Japanese children, but not the standard curves describing the distribution. AIM: The aim of the study was to construct a chart of body proportion of girls and boys in Japan. SUBJECTS AND METHODS: About 4% of the nationwide student population in Japan, more than 20,000 children per group of both boys and girls of all ages, was included randomly. The upper/lower ratio (ULR) was calculated from correlation tables containing stature and sitting height data reported in the Statistical Report on School Health and Hygiene. The subischial leg length was calculated by subtracting sitting height from stature. RESULTS: The present study showed that Japanese boys (13.5-17.5 years old) were taller and had relatively longer legs than Japanese girls in the same age group, and that the median ULRs of Japanese girls and boys (5.5-17.5 years old) were larger than the average of Caucasian girls and boys reported previously. In boys, the percentile values reached the lowest at 13.5 years, then increased slightly. On the other hand, in girls, the percentile values remained constant after 11.5 years, which is different in boys in the present study and in other Caucasian populations obtained from several studies. CONCLUSION: The new chart provides useful information in the assessment of the body proportions of Japanese children. PMID- 15799236 TI - Correcting physical activity energy expenditure for body size in children. AB - Being able to compare the energy cost of physical activity across and between populations is important. However, energy expenditure is related to body size, so it is necessary to appropriately adjust for differences in body size when comparisons are made. This study examined the relationship between the daily energy cost of activity and body weight in 47 children aged 6-10 years. Log-log regression showed weight1.0 to be an inappropriate adjustment for activity energy expenditure in children, with a more valid adjustment being weight0.3. Clearly, both weight dependent and non-weight dependent activities are part of everyday living in children. This balance influences how energy expenditure is correctly adjusted for body size. Investigators interpreting data of energy expenditure in children from children of different body sizes need to take this into consideration. PMID- 15799237 TI - Re: international definitions of overweight and obesity for children: a lasting solution? PMID- 15799238 TI - The proteasome. AB - The proteasome is an abundant multicatalytic enzyme complex present in the cytoplasm and nucleus of all eukaryotic cells. The primary function of the proteasome is to degrade proteins. While it was once thought to act primarily as a cellular "garbage disposal" that removed damaged or misfolded proteins from cells, the proteasome is now known to also remove various short-lived proteins that regulate the cell cycle, cell growth, and differentiation. By regulating the turnover of these proteins via timely degradation and recycling, the proteasome plays a critical role in the maintenance of cellular homeostasis. Substrates of the proteasome include cell-cycle regulators, signaling molecules, tumor suppressors, transcription factors, and antiapoptotic proteins; over 80% of all cellular proteins are recycled through the proteasome. This article discusses the structure and function of the proteasome, and its role in malignant cells and as a therapeutic target. PMID- 15799240 TI - Unraveling biologic therapy for Bcl-2-expressing malignancies. AB - Cancer cells that express excessive levels of Bcl-2 pose a major problem in the delivery of curative therapy. Most treatments for such cancer involve chemotherapy to induce the apoptotic process. While these therapies often result in disease control for periods of time, failure to initiate apoptosis as a result of acquired resistance limits the effectiveness of treatment for many common hematopoietic and solid malignancies, and ultimately death from the malignancy still occurs. Various anti-apoptotic proteins of the Bcl-2 family that localize to the mitochondria appear to be involved in this resistance mechanism. However, recent advances in the understanding of malignant cell biology, achieved through both genomics and proteomics, have made it possible to explore novel approaches directed at re-establishing sensitivity to chemotherapy, presenting an attractive strategy for cancer treatment. In this article we discuss how this may be achieved by lowering Bcl-2 anti-apoptotic protein expression using antisense oligonucleotides or, alternatively, by functionally antagonizing Bcl-2 using ligands of the mitochondrial benzodiazepine receptor. PMID- 15799239 TI - Mammalian target of rapamycin. AB - Targeted molecular therapeutics are tailored toward the genetic abnormalities that cause tumor progression. Modulation of certain signaling pathways that are aberrant in cancer cells has the potential to provide an effective, nontoxic approach to therapy in a broad range of cancers. Agents targeting BCR-ABL (imatinib mesylate [formerly known as STI-571], Gleevec; Novartis Pharmaceuticals Corp, East Hanover, NJ), retinoid receptor fusion proteins (all-trans retinoic acid), ErbB-2 or HER2/neu (trastuzumab, Herceptin; Genentech, Inc, South San Francisco, CA), epidermal growth factor receptor (IMC-C225 and ZD1839), and the phosphatidylinositol 3-kinase pathway (CCI-779) have all induced remarkable, nontoxic responses in a subset of patients with cancer and abnormalities in the corresponding signal transduction cascades. To achieve successful individualized therapy, the specific components within the aberrant signaling pathways that are driving the pathophysiology of the tumors must be identified in each patient. Molecular diagnostics can identify patients in whom the target is aberrant; linking molecular diagnostics with effective molecular therapeutics will be necessary to translate these concepts into approaches that will alter the outcome for patients with cancer. In addition, intermediary markers and/or molecular imaging techniques must be used to identify the biologically relevant dose that is sufficient to inhibit the target of interest. This review focuses on the P13K pathway, and novel molecules targeting this pathway, to illustrate the questions and challenges underlying the implementation of molecular therapeutics in breast and ovarian cancer. PMID- 15799242 TI - Clinical update: novel targets in multiple myeloma. AB - Novel therapies that can target the multiple myeloma (MM) cell, the MM cell patient bone marrow interaction, and the bone marrow milieu can overcome resistance to conventional therapy in preclinical models and clinical trials. Both proteasome inhibitor bortezomib and immunomodulatory drug Revimid (Celgene Corporation, Warren, NJ) have gone from laboratory bench to bedside in 3 to 4 years, and achieve responses even in patients with relapsed refractory MM. They are now undergoing clinical evaluation, alone and combined with conventional and novel therapies, for treatment of patients earlier in their disease course, and offer great promise to improve patient outcome in MM. PMID- 15799241 TI - Clinical update: novel targets in gynecologic malignancies. AB - The proteasome inhibitor bortezomib has shown activity in chemotherapy-resistant tumors and is approved for treatment of multiple myeloma. The critical component of bortezomib's antitumor activity is the inhibition of nuclear factor-kappa B (NF-kappaB). Patients with ovarian cancer respond to initial platinum-based chemotherapy, such as cisplatin. However, these agents have been shown to induce tumor cell survival by inducing NF-kappaB activity. Phase I trials of bortezomib in solid tumors, including ovarian cancer, are summarized and examined to determine if the compound can overcome the impact of chemoresistance. In one trial of single-agent bortezomib in advanced malignancies, it was deemed a safe and manageable drug with potential efficacy in solid tumors. A second phase I trial explored inhibition of NF-kappaB with bortezomib to see if the drug rendered platinum agents more sensitive in ovarian cancer patients. Seven of the nine patients in the study had major responses to the combination of carboplatin and bortezomib. The two trials indicate promising results for bortezomib in patients with solid tumors and patients with recurrent ovarian cancer, but further investigation is warranted. PMID- 15799243 TI - Airborne exposure to trihalomethanes from tap water in homes with refrigeration type and evaporative cooling systems. AB - This study evaluates airborne concentrations of common trihalomethane compounds (THM) in selected living spaces of homes supplied with chlorinated tap water containing >85 ppb total THM. Three small homes in an arid urban area were selected, each having three bedrooms, a full bath, and approximately 1000 square feet; two homes had standard (refrigeration-type) central air conditioning and the third had a central evaporative cooling system ("swamp cooler"). A high-end water-use pattern was used at each home in this exposure simulation. THM were concurrently measured on 4 separate test days in tap water and air in the bathroom, living room, the bedroom closest to the bathroom, and outside using Summa canisters. Chloroform (trichloromethane, TCM), bromodichloromethane (BDCM), and dibromochloromethane (DBCM) concentrations were quantified using U.S. EPA Method TO-14. The apparent volatilization fraction consistently followed the order: TCM > BDCM > DBCM. Relatively low airborne THM concentrations (similar to outdoors) were found in the living room and bedroom samples for the home with evaporative cooling, while the refrigeration-cooled homes showed significantly higher THM levels (three- to fourfold). This differential remained after normalizing the air concentrations based on estimated THM throughput or water concentrations. These findings indicate that, despite higher throughput of THM containing water in homes using evaporative coolers, the higher air exchange rates associated with these systems rapidly clears THM to levels similar to ambient outdoor concentrations. PMID- 15799244 TI - Diesel exhaust particulate matter dispersed in a phospholipid surfactant induces chromosomal aberrations and micronuclei but not 6-thioguanine-resistant gene mutation in V79 cells. AB - Diesel exhaust particulate material (DPM) was assayed for induction of chromosomal aberrations (CA), micronucleus (MN) formation, and 6-thioguanine resistant (TG9 gene mutation in V79 cells as a dispersion in dipalmitoyl phosphatidylcholine (DPPC) in physiological saline, a simulated pulmonary surfactant. Filter-collected automobile DPM provided for the study was not organic solvent extracted, but was directly mixed into DPPC in saline dispersion as a model of pulmonary surfactant conditioning of a soot particle depositing in a lung alveolus. A statistically significant difference was found between treated and control groups at all concentrations tested in a CA assay. Assay for MN induction also gave a positive response: Above 50 microg/ml, the frequencies of micronucleated cells (MNC) were about 2 times higher than those in the control group. The forward gene mutation assay did not show a positive response when cells were treated with up to 136 microg DPM/ml for 24 h, as dispersion in DPPC in saline. Some comparison assays were run on direct dispersions of the DPM into dimethyl sulfoxide, with results equivalent to those seen with a DPPC-saline preparation: DPM in dimethyl sulfoxide (DMSO) was positive for MN induction but was negative for forward gene mutation in V79 cells. The positive clastogenicity results are consistent with other studies of DPM dispersed into DPPC-saline surfactant that have shown activity in mammalian cells for sister chromatid exchange, unscheduled DNA synthesis, and MN induction. The forward gene mutation negative results are consistent with studies of that assay applied to V79 cells challenged with DPM solvent extract. PMID- 15799245 TI - JWA--a novel environmental-responsive gene, involved in estrogen receptor associated signal pathway in MCF-7 and MDA-MB-231 breast carcinoma cells. AB - The pyrethroid insecticide fenvalerate and the organophosphorus insecticide phoxim are now the most widely used agents for indoor pest control in China. Fenvalerate was shown to mimic estrogenic activity, whereas phoxim did not induce similar effects. Our previous studies demonstrated that JWA, a novel retinoic acid-inducible and cytoskeleton-associated gene, is also a potential environmental-responsive gene with increased expression to oxidative and heat shock stresses. In the present study, the influence of both fenvalerate and phoxim was examined on the expression of JWA in MCF-7 (ER+) and MDA-MB-231 (ER-) human breast carcinoma cell lines. Concentrations of 0.01, 1, and 100 micromol/L of fenvalerate or phoxim were selected to treat both MCF-7 and MDA-MB-231 cells at 1, 3, and 5 d, respectively. The MTT results only showed that fenvalerate stimulated MCF-7 cell proliferation. Western blot assay was employed to detect the expressions of JWA and heat-shock proteins (hsp27 and hsp70). The results showed that after treatment with fenvalerate, both JWA and hsp70 showed similar expression patterns in the both cell lines; however, all the expression patterns of JWA, hsp27, and hsp70 were evidently reversed between ER+ and ER- cells. In addition, phoxim-treated cells showed a concentration-dependent relationship in JWA expression at all time points. These results suggest that JWA has similar functions with respect to hsp27 and hsp70, and might be a novel signal molecule in estrogen receptor-related signal transduction pathways in mammalian cells. PMID- 15799247 TI - Effects of PCB 126 on primary immune organs and thymocyte apoptosis in chicken embryos. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and polychlorinated biphenyl (PCB) 126 produce thymic atrophy and immunosuppression. This study explored the hypothesis that the thymic atrophy produced by developmental exposure to PCB 126 is associated with an increase in apoptotic thymocytes at the end of incubation in chicken embryos. Eggs were injected via the air cell with PCB 126 (0.05, 0.13, 0.32, 0.64, and 0.80 ng/g egg) on d 0 of incubation, and tissues were collected on d 20. Controls included noninjected and vehicle-injected (sunflower oil) eggs. Thymocytes were cultured for 6 h and analyzed by flow cytometry for decreased DNA content (propidium iodide staining) and cell size (forward scatter), which indicate apoptosis. PCB 126 induced dose-dependent mortality with an LD50 of 1.01 ng/g and lowest-observed-effect concentration (LOEC) of 0.32 ng/g. Teratogenic effects commonly associated with TCDD and planar PCBs, including cranial and foot deformities and subcutaneous edema, tended to increase with dose of PCB 126. PCB 126 reduced thymus mass by approximately 20% at 0.64 and 0.8 ng/g, the number of viable thymocytes by approximately 20-24% at and above 0.13 ng/g, and the number of bursal lymphoid cells by 57% at 0.64 ng/g. The percentage of apoptotic thymocytes increased with dose, reaching levels 2 times greater than controls at 0.8 ng/g. Electrophoresis of low-molecular-weight DNA from thymocytes of all doses demonstrated fragments in multiples of 180 bp. This DNA laddering is a hallmark of apoptosis. At all doses, thymocytes exhibited caspase-3 activation, another indicator of apoptosis. The results of this experiment supported the hypothesis that the thymic atrophy produced by developmental exposure to PCB 126 in chicken embryos is associated with an increase in apoptotic thymocytes on embryonic d 20. PMID- 15799248 TI - [Strengthening the mother-child relationship, and its impact on substance misuse therapy]. PMID- 15799246 TI - Does high organochlorine (OC) exposure impair the resistance to infection in polar bears (Ursus maritimus)? Part II: Possible effect of OCs on mitogen- and antigen-induced lymphocyte proliferation. AB - Previous studies have reported alarmingly high levels of organochlorines (OCs), particularly polychlorinated biphenyls (PCBs), in free-ranging polar bears (Ursus maritimus). In this study plasma concentration of PCBs ranged from 14.8 to 200 ng/g wet weight. The aim of the study was to investigate associations between OCs and lymphocyte proliferation after in vitro stimulation with different mitogens and antigens. In 1998 and 1999, 26 and 30 free-ranging polar bears from Svalbard and Churchill, Canada, respectively, were recaptured 32-40 d following immunization with inactivated tetanus toxoid and hemocyanin from keyhole limpets (KLH) to sensitize lymphocytes. At recapture, blood was sampled for determination of plasma levels of PCBs and organochlorine pesticides (OCPs) and lymphocyte proliferation after in vitro stimulation with specific mitogens- phytohemagglutinin (PHA), pokeweed mitogen (PWM), concanavalin A (Con A), lipopolysaccharide (LPS), and purified protein derivative of Mycobacterium avium subsp. paratuberculosis (PPD)--and antigens: tetanus toxoid and KLH. The combinations of sum(PCBs) (sum of 12 individual PCB congeners), sum(OCPs) (sum of 6 OCPs), and their interactions contributed up to 15% of the variations in the lymphocyte responses. By using multiple regression analyses, followed by classical mathematic function analyses, thresholds for immunomodulation were estimated. Depending on the lymphocyte proliferation response studied, the estimated thresholds for significant immunomodulation were within the concentration ranges 32-89 ng/g wet weight (ww) and 7.8-14 ng/g ww for sum(PCBs) and sum(OCPs), respectively. Thus, this study demonstrated that OC exposure significantly influences specific lymphocyte proliferation responses and part of the cell-mediated immunity, which also is associated with impaired ability to produce antibodies (Lie et al., 2004). PMID- 15799249 TI - [Are Alzheimer's drugs effective?]. PMID- 15799250 TI - [Prioritizing instead of waiting in line]. PMID- 15799251 TI - [HLA and infections]. PMID- 15799252 TI - [Radiology supporting the treatment of back pain]. PMID- 15799253 TI - [Follow-up at home for children under oral anticoagulant treatment]. PMID- 15799254 TI - [Buprenorphine treatment during pregnancy--risk factors and advantages need to be considered carefully]. PMID- 15799255 TI - [Update on current care guidelines. Headache among children]. PMID- 15799256 TI - [Metabolic acidosis quickly resolved]. PMID- 15799257 TI - [ Duodecim's comment on circumcision of boys]. PMID- 15799258 TI - [Confusing conclusion about circumcision]. PMID- 15799259 TI - [Duodecim society and ritualistic circumcision]. PMID- 15799260 TI - [More comments on circumcision of boys]. PMID- 15799261 TI - [Series of current care recommendation articles are reaching more popularity]. PMID- 15799262 TI - [Ice fishing]. PMID- 15799263 TI - [Cold effects our health in many ways]. PMID- 15799264 TI - [Exposure to cold and the symptoms thereof]. PMID- 15799266 TI - [Cold, hot and mortality]. PMID- 15799265 TI - [Response to cold exposure and physiology of the body]. PMID- 15799267 TI - [How does frost affect breathing? Asthma and pulmonary congestion in cold climate]. PMID- 15799268 TI - [Cold and infections]. PMID- 15799269 TI - [Frostbite and other problems of skin expose to cold]. PMID- 15799270 TI - [Any advantage of cold and cryotherapy?]. PMID- 15799272 TI - Training. SHAs take on NHSU role. AB - Most of the functions of the NHSU, the corporate training body for NHS staff, are expected to be devolved to strategic health authorities, according to a report by a 'transition team' that is due to land on ministers' desks this weeks. Meanwhile, an e-mail to staff from departing NHSU chief executive Professor Bob Fryer claims that 'not everyone wanted NHSU to succeed'. PMID- 15799273 TI - Public health. A weight on PCTs' minds: how to get LDPs into shape on time. AB - The public health delivery paper tells the service how to set about achieving the goals laid out in November's white paper. But with only weeks to go before PCTs must submit their local delivery plans to SHAs and worries about the lack of ringfenced funding, the pressure is on. We look at the problems PCTs face and detail the key areas for action. PMID- 15799274 TI - On treating patients well. PMID- 15799276 TI - Barometer. Acute trusts February 2005. AB - Almost two thirds of acute trusts rate the quality of commissioning from their PCTs at three out of 10 or less, according to the latest HSJ Barometer survey. This is the lowest score since we began the survey a year ago. Confidence in their performance against the 98 per cent four-hour A&E target fell sharply from a December high to an average of 6.87. The survey also found that fewer than one in seven trusts were confident of winning foundation status by the end of 2006 07. PMID- 15799275 TI - The HSJ debate. Self-management of chronic disease doesn't work. AB - The policy focus on self-management of long-term conditions is misplaced and not supported by good evidence, acccording to Professor Mike Bury, author of the new book Health and Illness. In a debate with David Pink of the Long Term Medical Conditions Alliance, he also argues that the levels of chronic disease in the U.K. are likely to have been exaggerated. PMID- 15799277 TI - Mental health. Respect your elders. AB - Mental health services for older people tend to be neglected by policy in favour of general adult mental health services. Inadequate staffing, lack of capacity and insufficient intermediate care are all contributing factors. Salford and Trafford trust now has accredited training for primary care nurses in the identification and initial management of mental health problems in older people. PMID- 15799278 TI - On mental health. PMID- 15799279 TI - On clinical management. PMID- 15799280 TI - Regional focus: South East. Out of the city. PMID- 15799281 TI - Executive salaries. Because you're worth it? PMID- 15799283 TI - On mobile health. PMID- 15799282 TI - Training. Stage school. PMID- 15799284 TI - Choose and book. By the book. AB - The National Audit Office predicts that 60-70 per cent of hospitals and GP practices will be linked to the electronic booking service by the end of the year. By the end of last year the system had seen just 63 referrals, despite an earlier target of 250,000. Various surveys continue to show strong suspicion and lack of knowledge among GPs. PMID- 15799285 TI - National programmes. State of the nations. AB - Besides some centralised procurement, Scotland's IT programme is being implemented on a bottom-up basis. Wales has adopted a less rigid approach than England, encouraging stakeholder involvement. Northern Ireland's programme is more centralised, with all its hospital IT systems in two service centres. PMID- 15799286 TI - Information workforce. Tomorrow people. Interview by Steve Mathieson and Nick Edwards. PMID- 15799287 TI - Outside edge. National program for IT. PMID- 15799288 TI - The 26.2-mile security rule: is your organization gaining on compliance or just running in place? PMID- 15799289 TI - HIPAA marches on--or does it? PMID- 15799290 TI - HIPAA audit and system activity review. Developing a process that focuses on the greatest risks first. PMID- 15799291 TI - Real-world RHIO. A regional health information organization blazes a trail in Upstate New York. PMID- 15799292 TI - Project management 101: skills for leading and working in teams, Part 3. PMID- 15799293 TI - Ready for the EHR? A new survey measures EHR implementation and individual readiness. PMID- 15799294 TI - Safeguarding ePHI from fire the dry way. PMID- 15799295 TI - The CCR--not an EHR. PMID- 15799296 TI - Reporting security incidents. PMID- 15799297 TI - Getting to know PubMed: an overview. PMID- 15799298 TI - Data linkage--ready or not, it's here. PMID- 15799299 TI - The trouble with DRGs: Part 2. PMID- 15799300 TI - New procedures for joint resurfacing of the knee. PMID- 15799301 TI - The backbone of the security rule. PMID- 15799302 TI - [Differential diagnosis of COPD and asthma]. PMID- 15799303 TI - [Recent progress in leukotriene research]. PMID- 15799304 TI - [The characteristic of the pollen dispersal of Cryptomeria japonica]. PMID- 15799305 TI - [Development of patient satisfaction questionnaire in allergic rhinitis]. AB - Recent suggestions emphasize the importance of assessing medical care outcomes from the point of view of both professional opinion and patient's subject like patient satisfaction as well as quality of life. Self-administered questionnaires are often used as an instrument for this purpose. When compared with quality of life, there is less useful questionnaire for patient satisfaction. Accordingly, we here aimed to develop the questionnaire, which is disease-specific, comprehensive, simple, clinically available and satisfied psychometric validation study. The items were collected from patients and satisfaction-related literature's review, and the total 20 were selected finally from them after excluding inadequate or overlapped items. Then, we conducted validation study in 214 subjects sampled randomly from 603 patients with Japanese cedar pollinosis who had visited clinics in 2003 by mailing self-administered questionnaire in spring, 2004. The response rate was 65.3%. In factor analysis, items were divided into four domains such as doctor-patient relationship, treatment outcome, geniality in hospital and convenience of medical service. The former two domains were the most correlated well with patient satisfaction. The quality of this questionnaire such as responsiveness, reliability, construct validity, convergent/discriminate validity, and clinical validity satisfied international standard. Thus, it was verified that our newly developed questionnaire is a useful and scientific tool for examination of patient satisfaction with medical care in allergic rhinitis. PMID- 15799307 TI - [Serum rheumatoid factor and peripheral blood eosinophil counts in patients with bronchial asthma]. AB - These days some reports say that bronchial asthma, especially severe asthma, is systemic inflammatory and its severity correlates with eosinophil counts. Some patients with asthma have positive tests for rheumatoid factor (RF) which is detected in systemic hyper immunoreactive desease such as rheumatoid arthritis. So we investigated each of these factors in asthmatic patients. We randomly selected 100 asthmatics from regular outpatients of our hospital. In a positive RF level group (over 21 IU/ml), Eosinophil counts were significantly higher than those in a negative RF level group. RF levels were significantly and positively correlated with the logarithm of eosinophil count. And in severe groups (patients with severe asthma, treated with high dose inhaled steroids, and with history of systemic steroids use for the last one month), RF levels were significantly higher than those in other groups. Our result suggested that RF levels reflect eosinophilia and asthma severity. PMID- 15799306 TI - [Effects of dexamethasone on apoptosis of eosinophils infiltrated into bronchoalveolar lavage fluid after Sephadex bead treatment in rat]. AB - Eosinophilic inflammation of the airways is a key characteristic of asthma. Glucocorticoids can suppress the inflammatory response in part by promotion of eosinophilic apoptosis. We investigated the effects of glucocorticoids on leukocyte infiltration and apoptotic resolution of eosinophils and neutrophils in Sephadex-treated rat lung. Sephadex beads were injected intravenously, followed 24 h later by i.p. administration of dexamethasone (DEX, 0.1 mg/kg) or its vehicle. At 24 h post-DEX treatment, the bronchoalveolar lavage fluid (BALF) was collected. Differential leukocyte counts and the numbers of apoptotic eosinophils and neutrophils, and macrophages with engulfed eosinophils or neutrophils in BALF were determined microscopically from Diff-Quik stained cytospin preparations. Sephadex beads markedly increased cell counts of eosinophils and neutrophils in BALF. Compared with a vehicle-treated group, the DEX treatment significantly decreased the number of eosinophils, but not neutrophils, in BALF. Dexamethasone in BALF also significantly increased eosinophilic apoptosis and engulfment of apoptotic eosinophils by macrophages, but had no effect on neutrophilic apoptosis and engulfment of apoptotic neutrophils by macrophages. These results suggest that the increased clearance of eosinophils from airways by glucocorticoids may be partly due to the promotion of eosinophilic apoptosis. PMID- 15799308 TI - [Classification and diagnosis of rheumatoid arthritis]. PMID- 15799309 TI - [Epidemiology of rheumatoid arthritis in Japan]. PMID- 15799310 TI - [Course of rheumatoid arthritis]. PMID- 15799311 TI - [Predictive factors in prognosis of rheumatoid arthritis]. PMID- 15799312 TI - [Clinical epidemiology of rheumatoid arthritis--activities of J-ARAMIS]. PMID- 15799313 TI - [K/BxN mouse]. PMID- 15799314 TI - [Collagen-induced arthritis (CIA)]. PMID- 15799315 TI - [SKG mice as a model of RA]. PMID- 15799316 TI - [Antibody-induced arthritis]. PMID- 15799317 TI - [Adjuvant-induced arthritis]. PMID- 15799318 TI - [IL-1 receptor antagonist knockout mice]. PMID- 15799319 TI - [SCID-HuRAg mouse]. PMID- 15799320 TI - [gp130F759 mouse]. PMID- 15799321 TI - [Structure and function of the synovial joint]. PMID- 15799322 TI - [Autoimmune abnormality]. PMID- 15799323 TI - [Involvement of osteoclasts in cartilage and bone destruction in rheumatoid arthritis]. PMID- 15799324 TI - [Immune and skeletal systems]. PMID- 15799325 TI - [Histopathological study on the process of joint destruction in rheumatoid arthritis]. PMID- 15799326 TI - [Role of apoptosis in the pathogenesis of rheumatoid arthritis]. PMID- 15799327 TI - [Nitric oxide in rheumatoid arthritis]. PMID- 15799328 TI - [Current studies on susceptibility genes in rheumatoid arthritis]. PMID- 15799329 TI - [HLA-DRB1 gene for rheumatoid arthritis]. PMID- 15799330 TI - [Susceptibility genes in rheumatoid arthritis: mutation in DR3 gene]. PMID- 15799331 TI - [Susceptibility genes in rheumatoid arthritis: peptidylarginine deiminase (PADI) type 4]. PMID- 15799332 TI - [Environmental exposures associated with rheumatoid arthritis]. PMID- 15799333 TI - [Role of prostaglandins in rheumatoid arthritis]. PMID- 15799334 TI - [Bradykinin--with special reference to its action on nociceptors]. PMID- 15799336 TI - [Interleukin-1 in pathogenesis of rheumatoid arthritis]. PMID- 15799335 TI - [Tumor necrosis factor alpha and beta (TNF-alpha and -beta) in pathogenesis of rheumatoid arthritis]. PMID- 15799337 TI - [Interleukin-6 in pathogenesis of rheumatoid arthritis]. PMID- 15799338 TI - [Interleukin-8 in pathogenesis of rheumatoid arthritis]. PMID- 15799340 TI - [Interleukin-16 in pathogenesis in rheumatoid arthritis]. PMID- 15799341 TI - [Interleukin-18 in pathogenesis of rheumatoid arthritis]. PMID- 15799339 TI - [Interleukin-15 in pathogenesis of rheumatoid arthritis]. PMID- 15799342 TI - [Interferon-gamma in pathogenesis of rheumatoid arthritis]. PMID- 15799343 TI - [Osteopontin in rheumatoid arthritis]. PMID- 15799345 TI - [Role of chemokines in the pathogenesis of rheumatoid arthritis]. PMID- 15799344 TI - [Angiogenic growth factors in rheumatoid arthritis]. PMID- 15799346 TI - [Adhesion molecule in pathogenesis of rheumatoid arthritis]. PMID- 15799348 TI - [Systemic features of rheumatoid arthritis]. PMID- 15799347 TI - [Proteinases in pathogenesis of rheumatoid arthritis]. PMID- 15799349 TI - [Joint involvement in rheumatoid arthritis]. PMID- 15799350 TI - [Dermatologic conditions & nail manifestations in rheumatoid arthritis]. PMID- 15799351 TI - [Rheumatic diseases and ocular features]. PMID- 15799352 TI - [Mental status in patients with rheumatism]. PMID- 15799353 TI - [Pleuropulmonary manifestations in patients with rheumatoid arthritis]. PMID- 15799354 TI - [Renal disease in rheumatoid arthritis]. PMID- 15799355 TI - [Rheumatoid heart lesions]. PMID- 15799356 TI - [Gastrointestinal manifestations in rheumatoid arthritis]. PMID- 15799357 TI - [Spinal disorders associated with rheumatoid arthritis]. PMID- 15799358 TI - [Neuromuscular involvement in rheumatoid arthritis]. PMID- 15799359 TI - [Reticuloendotherial system involvement in rheumatoid arthritis]. PMID- 15799360 TI - [Rheumatoid vasculitis]. PMID- 15799362 TI - [Basic evaluation of patients with arthritis rheumatoid: history and physical examination]. PMID- 15799361 TI - [Secondary amyloidosis associated with rheumatoid arthritis (RA)]. PMID- 15799364 TI - [Disease activity and clinical examination in rheumatoid arthritis]. PMID- 15799363 TI - [Progress in early diagnosis of rheumatoid arthritis]. PMID- 15799365 TI - [Simple hematological blood tests and urine and stool tests for diagnosis of rheumatoid arthritis]. PMID- 15799366 TI - [Serum biochemical values in rheumatoid arthritis]. PMID- 15799367 TI - [Inflammatory markers in rheumatoid arthritis]. PMID- 15799368 TI - [Immunological and serological tests for diagnosis of rheumatoid arthritis- introduction]. PMID- 15799370 TI - [Immunological and serological tests for diagnosis of rheumatoid arthritis: IgM RF]. PMID- 15799369 TI - [Immunological and serological tests for diagnosis of rheumatoid arthritis: MMP 3]. PMID- 15799371 TI - [Immunological and serological tests for diagnosis of rheumatoid arthritis: IgG RF]. PMID- 15799372 TI - [Immunological and serological tests for diagnosis of rheumatoid arthritis: anti agalactosyl IgG antibody]. PMID- 15799373 TI - [immunological and serological tests for diagnosis of rheumatoid arthritis: antinuclear antibodies]. PMID- 15799374 TI - [Anti-cyclic citrullinated peptide antibody as a serological marker for the diagnosis of rheumatoid arthritis]. PMID- 15799375 TI - [Anti-calpastatin antibody as a serological marker for the diagnosis of rheumatoid arthritis]. PMID- 15799376 TI - [Antifilaggrin antibody as a serological marker for the diagnosis of rheumatoid arthritis]. PMID- 15799377 TI - [Anti-type II collagen autoantibodies in rheumatoid arthritis]. PMID- 15799378 TI - [Laboratory tests of coagulation and fibrinolysis for the diagnosis of rheumatoid arthritis]. PMID- 15799379 TI - [Significance of synovial fluid analysis in the diagnosis of rheumatoid arthritis]. PMID- 15799380 TI - [Histopathological examination of rheumatoid arthritis by biopsy]. PMID- 15799381 TI - [Pulmonary function tests in patients with rheumatoid arthritis]. PMID- 15799382 TI - [Renal function tests in patients with rheumatoid arthritis]. PMID- 15799383 TI - [Electrodiagnosis of neuropathy and myopathy in rheumatoid arthritis]. PMID- 15799384 TI - [Salivary gland examinations in patients with rheumatoid arthritis]. PMID- 15799385 TI - [Abnormalities in electrocardiogram of patients with rheumatoid arthritis]. PMID- 15799386 TI - [Radiographic imaging in rheumatoid arthritis]. PMID- 15799387 TI - [CT for diagnosis of rheumatoid arthritis]. PMID- 15799388 TI - [Ultrasonographic assessment of joints in rheumatoid arthritis]. PMID- 15799389 TI - [Magnetic resonance imaging in rheumatoid arthritis]. PMID- 15799390 TI - [Bone densitometry in patients with rheumatoid arthritis]. PMID- 15799391 TI - [Scintigraphy in rheumatoid arthritis]. PMID- 15799392 TI - [Diagnostic arthroscopy for rheumatoid arthritis]. PMID- 15799393 TI - [Special techniques in differentiating RA from the remaining confusing connective tissue diseases]. PMID- 15799395 TI - [Guidelines for RA therapies]. PMID- 15799394 TI - [Lesson from ACR guidelines for management of RA]. PMID- 15799397 TI - [Strategy for the management of RA patients]. PMID- 15799396 TI - [Guidelines for usage of biological agents for treatment of rheumatoid arthritis]. PMID- 15799398 TI - [Treatment of juvenile idiopathic arthritis]. PMID- 15799399 TI - [Psychosomatic therapy of chronic pain in patients with rheumatoid arthritis]. PMID- 15799400 TI - [Osteoporosis in RA: pathophysiology and treatment]. PMID- 15799401 TI - [Corticosteroids for treatment of rheumatoid arthritis]. PMID- 15799402 TI - [NSAIDs for therapy of rheumatoid arthritis. Introduction]. PMID- 15799404 TI - [Disease modifying anti-rheumatic drugs (DMARDs): an overview]. PMID- 15799403 TI - [Selective COX-2 inhibitors for therapy of rheumatoid arthritis]. PMID- 15799405 TI - [Leflunomide for therapy of rheumatoid arthritis]. PMID- 15799406 TI - [Methotrexate for therapy of rheumatoid arthritis]. PMID- 15799407 TI - [T-614 for therapy of rheumatoid arthritis]. PMID- 15799408 TI - [Tacrolimus for therapy of rheumatoid arthritis]. PMID- 15799410 TI - [Infliximab for therapy of rheumatoid arthritis]. PMID- 15799409 TI - [Biological agents for therapy of rheumatoid arthritis--overview]. PMID- 15799411 TI - [Etanercept for therapy of rheumatoid arthritis]. PMID- 15799412 TI - [Adalimumab for therapy of rheumatoid arthritis]. PMID- 15799413 TI - [Anakinra for therapy of rheumatoid arthritis]. PMID- 15799414 TI - [MRA for therapy of rheumatoid arthritis]. PMID- 15799415 TI - [CD20 antibody for treatment of rheumatoid arthritis]. PMID- 15799416 TI - [Combination drug therapy in rheumatoid arthritis]. PMID- 15799418 TI - [Adverse effects of NSAIDs in treatment of rheumatoid arthritis]. PMID- 15799417 TI - [Adverse effects of corticosteroids in treatment of rheumatoid arthritis]. PMID- 15799419 TI - [Adverse effects of DMARDs in treatment of rheumatoid arthritis]. PMID- 15799421 TI - [Intra-articular injection for rheumatoid arthritis]. PMID- 15799420 TI - [Adverse effects of biologic agents in treatment of rheumatoid arthritis- prevention of tuberculosis]. PMID- 15799423 TI - [Therapeutic exercise in rheumatoid arthritis]. PMID- 15799422 TI - [Efficacy of clinical path in the treatment of patients with rheumatoid arthritis]. PMID- 15799424 TI - [Occupational therapy for rheumatoid arthritis]. PMID- 15799425 TI - [Physical therapy for rheumatoid arthritis]. PMID- 15799426 TI - [Splints and orthoses for treating rheumatoid arthritis]. PMID- 15799428 TI - [Surgical treatment for upper extremity disorders in rheumatoid arthritis]. PMID- 15799427 TI - [Role of Rheumatologists and medical services for rheumatoid arthritis in Japan]. PMID- 15799429 TI - [Surgical treatment for the lower limb in rheumatoid arthritis]. PMID- 15799430 TI - [Rheumatoid arthritis of the cervical spine]. PMID- 15799431 TI - [Short term (four weeks) in-patient rheumatoid arthritis (RA) education program for teaching patients how to cope with RA]. PMID- 15799432 TI - [Collaboration among hospitals and clinics for the long-term management of patients with rheumatoid arthritis]. PMID- 15799433 TI - [Social psychological problems in patients with functional disability from rheumatoid arthritis]. PMID- 15799434 TI - [Holistic medicine for treatment of rheumatoid arthritis]. PMID- 15799435 TI - [Apheresis treatment for rheumatoid arthritis]. PMID- 15799436 TI - [Leukocytapheresis for therapy of rheumatoid arthritis]. PMID- 15799437 TI - [Gene therapy for arthritis--past and future]. PMID- 15799438 TI - [Potential application of oligodeoxynucleotides for rheumatoid arthritis]. PMID- 15799439 TI - [Regenerative medicine for rheumatoid arthritis--current status and problems]. PMID- 15799440 TI - [Cell transplantation for therapy of rheumatoid arthritis]. PMID- 15799441 TI - [Factors for bone and cartilage regeneration]. PMID- 15799442 TI - [Repair of arthritic lesions using cultured mesenchymal cells and artificial joints]. PMID- 15799443 TI - [Tailor-made medicine and genomic drug discovery in rheumatoid arthritis]. PMID- 15799444 TI - [Robotic systems in orthopedic surgery for rheumatoid arthritis]. PMID- 15799445 TI - [Support system for medical examination and treatment and for clinical research in rheumatic diseases]. PMID- 15799446 TI - [Rehabilitation and long-term care insurance: rheumatoid arthritis]. PMID- 15799447 TI - [Anti-rheumatoid biologics and pharmacoeconomic evaluation]. PMID- 15799448 TI - [Potential involvement of cellular senescence in the pathogenesis of bone and joint disease]. PMID- 15799450 TI - Effect of cooking on the loss of persistent organic pollutants from salmon. AB - Recent studies have raised concern over the presence of high levels of persistent organic pollutants (POPs) in farmed fish relative to wild specimens of the same species, particularly salmon. Although cooking is known to reduce the burden of POPs in fish, the mechanisms of loss/degradation are not clearly understood. This study investigated the loss of POPs, including polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), p,p'-DDT [2,2-bis(p-chlorophenyl)-1,1,1 trichloroethane] and its related metabolites (sum noted as DDTs), and chlordane congeners, from salmon (Salmo salar) steaks when subjected to baking, boiling, frying, or microwave cooking. Ranges in the raw flesh were 25.1-62.9 ng/g wet weight (ww) for PCBs, 2.5-7.6 ng/g ww for PBDEs, 2.4-5.3 ng/g ww for chlordanes, and 17.5-43.8 ng/g ww for DDTs. Analysis of raw steaks from along the fish body revealed a significant variation of POP concentrations along the fish body, with higher concentrations at the head end than the tail, with a peak in the central section. After cooking, levels of POPs decreased in salmon steak with an average loss of 26 +/- 15% relative to the initial POP load in the raw steak. The removal of the skin from the cooked salmon steak resulted in a further average loss of 9 +/- 3%. The loss of POPs did not differ significantly between cooking methods. Losses of POPs were significantly and linearly correlated with the losses of lipid during cooking, suggesting removal of lipids is the critical factor for POPs reduction in cooked fish. Cooking of raw fish contaminated with POPs can therefore be expected to reduce the consumption exposure risk to human health. PMID- 15799449 TI - Estrogenic activity of cosmetic components in reporter cell lines: parabens, UV screens, and musks. AB - In this work, the estrogenic effects of three classes of substances included in cosmetic formulations-parabens, ultraviolet (UV) screens, and musk fragrances were studied. Their estrogenic activity was measured with the use of three reporter cell lines: HELN, HELN ERalpha, and HELN ERbeta. These three cell lines allowed for the measurement of estrogenic activity toward estrogen receptors alpha and beta (ERalpha and ERbeta, while taking nonspecific interactions into account. Eight of the 15 substances tested showed specific estrogenic activity with the following degree of potency on ERalpha butylparaben > propylparaben > homosalate = octyl-dimethyl-PABA = 4-methyl-benzylidenecamphor = octyl methoxycinnamate > ethylparaben = galaxolide. Among these active substances, parabens activated ERalpha and ERbeta similarly, UV screens activated ERalpha moderately and had almost no effect on ERbeta, and fragrances did not activate ERbeta. Methylparaben, ethylparaben, musk moskene, celestolide, and cashmeran did not activate estrogenic responses up to 10(-5) M. Musk ketone and benzophenone-3 were not considered estrogenic at 10(-5) M. PMID- 15799451 TI - Oxidative stress and sex steroid levels in fish following short-term exposure to pulp-mill effluents. AB - This study investigates the relationship between oxidative stress and reproductive dysfunction in wild white sucker (Catostomus commersoni) with short term exposures to pulp-mill effluent. Hepatic oxidative damage, as quantified using 2-thiobarbituric acid-reactive substances (TBARS), was often increased with effluent exposure within 4-8 d, but responses varied by species, sex, and effluent. Fatty acyl-coenzyme A (CoA) oxidase (FAO) and ethoxyresorufin O deethylase (EROD) activities were also significantly induced between 4 and 8 d of exposure. There were marked species differences in oxidative stress, as TBARS, FAO, and EROD responses in white sucker differed dramatically from those of longnose sucker (Catostomus catostomus) exposed under identical conditions. Exposure for 8 d to pulp-mill effluent delayed ovulation in white sucker, and these delays were independent of changes in circulating testosterone and 17alpha,20beta-dihydroxy-4-pregnen-3-one titers. Evaluations of the effects of pulp mill effluent on in vivo plasma steroid levels and in vitro steroidogenic capacities were compromised due to caging stress. In vivo preexposure to pulp mill effluent did not reduce in vitro ovarian follicle steroidogenic capacities when exposed to additional reactive oxygen species (ROS) generators. Endocrine and oxidative stress parameters may be interrelated, as the in vivo administration of ferric nitrilotriacetate (Fe(3+)NTA) significantly reduced circulating sex steroids. Administration of a superactive GnRH analog containing a dopamine inhibitor significantly increased TBARS within 24 h, indicating endocrine status is capable of modifying oxidative stress responses. This study provides new knowledge regarding the onset of oxidative stress and changes in reproductive endpoints in fish following pulp-mill effluent exposure. PMID- 15799452 TI - Characterization of the metabolic interaction between trihalomethanes and chloroacetic acids using rat liver microsomes. AB - The aim of this study was to investigate the in vitro metabolism of trihalomethanes (THMs) in the presence of trichloroacetic acid (TCA), dichloracetic acid (DCA), monochloroacetic acid (MCA), and 4-methylpyrazole (4 MP) using liver microsomes from male Sprague-Dawley rats. Using the vial equilibration technique, initial experiments were carried out with starting concentrations of approximately 40 ppm THMs and 12-22 mM chloroacetic acids. The results indicated a mutual metabolic inhibition between THMs present as binary or quaternary mixtures. Although DCA and MCA had no influence on THMs, TCA produced a marked inhibition of the metabolism of all THMs: chloroform (CHCl3) (55%), bromodichloromethane (BDCM) (34%), dibromochloromethane (DBCM) (30%), and bromoform (TBM) (23%). The presence of 4-MP also reduced THM metabolism, the importance of which decreased in the following order: CHCl3 > BDCM > DBCM = TBM. In further vial equilibration experiments, using 9-140 ppm as starting concentrations of THMs, enzyme kinetic parameters (i.e., Michaelis constant, K(m), and maximum velocity, V(max)) were determined both in the absence and in the presence of TCA (12.2 mM). Results are consistent with a competitive inhibition between TCA and CHCl3, whereas the metabolic inhibition of BDCM and TMB by TCA was non-competitive. As for DBCM, results suggest a more complex pattern of inhibition. These results suggest that CYP2E1 is involved in the metabolism of THMs as well as in the metabolic interaction between THMs and TCA. PMID- 15799453 TI - Metabolism of territrem B and C in liver microsomes from 14-wk-old Wistar rats is catalyzed by cytochrome P-450 3A. AB - The metabolism of territrem B (TRB) and territrem C (TRC) in liver microsomes of 14-wk-old male and female Wistar rats was investigated. Metabolism of TRB to 4beta-hydroxylmethyl-4beta-demethylterritrem B (MB2), O-demethylation of the methoxy group of the aromatic moiety of TRB to form MB4 (same structure as TRC), and metabolism of TRC to 4beta-hydroxylmethyl-4beta-demethylterritrem C (MC) were observed in both genders. However, the amounts of MB2, MB4, and MC formed in females were much lower than in males. To investigate which cytochrome P-450 (CYP450) isoforms were involved in each step, four CYP450 isotype-specific inhibitors (furafylline, orphenadrine, cimetidine, and troleandomycin) and antibodies against CYP1A1, CYP2B1, CYP2C11, or CYP3A2 were used. Formation of MB2, MB4, and MC was markedly inhibited by cimetidine and troleandomycin, but less by furafylline and orphenadrine. Anti-CYP3A2 antibody completely inhibited MB, MB, and MC formation, while antibodies against CYP1A1, CYP2B1, or CYP2C11 produced no marked effect. Of the seven tested supersomes from baculovirus transformed insect cells expressing rat CYP450 isoforms (1Al, 1A2, 2B1, 2C11, 2C12, 3A1, and 3A2), only those expressing CYP3A1 and CYP3A2 metabolized TRB and TRC. The amounts of MB2, MB4, and MC formed by male and female rat liver microsome preparations were related to the testosterone 6beta-hydroxylase activity and CYP3A1/2 protein content of the preparation. Immunoblotting showed that CYP3A1 was expressed in both genders, but at different levels, while CYP3A2 was only expressed in males. These results suggest that the formation of MB2, MB4, and MC in liver microsomes from 14-wk-old rats of either gender is mediated by CYP3A1 and CYP3A2. PMID- 15799454 TI - Key issues relating to decentralization at the provincial level of health management in Cambodia. AB - The following study was conducted as part of a review of management systems at a Provincial Health Department (Kampong Cham Province) and a National Health Programme (National Immunization Program) in 2002-2003 in Cambodia. The aim of this paper is to identify, analyse and recommend those management system factors that are critical to the success of health management performance, with a particular focus on provincial management. The review has identified critical success factors associated with health management performance at the sub-national level that include a stronger role for effective human resource management in health sector reform, elevation of the status of planning in senior level management, and the development of a more comprehensive and transparent finance system. These success factors will position the provincial level of health management to respond more effectively to the reform challenges of administrative de-concentration and political decentralization that are currently underway across a range of government sectors in Cambodia. PMID- 15799455 TI - Decentralizing EPI services and prospects for increasing coverage: the case of Tanzania. AB - Primary health Care (PHC) strategies were adopted widely in 1978 after the Alma Ata declaration to improve accessibility to health services and the health of the people. Of the strategies of PHC was the decentralization of health services to lower levels in order to enhance participation and responsiveness of the health system to local problems. While PHC was being promoted vertical programmes such as the expanded programme on immunization (EPI) were also being promoted and achieved substantial benefits. However, almost 25 years later many countries have not been able to achieve these health goals. This study addressed the question: Can we make the process of health care decentralization more likely to support health system and EPI goals? This study analysed the experience of EPI decentralization at national, regional and district levels. Several stakeholders were identified who were supportive and others who were non-supportive of the process. Community support to EPI measured by using willingness to pay (WTP) for kerosene (to keep vaccines cool) was low. It was significantly (p < 0.05) associated with whether providers in the nearest health facility properly attended the target population and whether the providers in the facility were available when needed. There was a substantial stakeholder support and opposition to the process of decentralization at the district level. Community support was not high possibly due to the perceived non-availability of the service providers and their lack of awareness of the population they serve. It was proposed that reforms should give priority to the involvement of communities and peripheral health facility providers in the process. PMID- 15799456 TI - Inequalities in self-reported health within Spanish Regional Health Services: devolution re-examined? AB - There is evidence of small inter-regional inequalities in both health and health expenditure both before and after the decentralization of the Spanish National Health Service (Spanish NHS). However, it still has not been established whether devolution has exerted any influence on the development of intra-regional inequalities in health. This study examines the existence of socioeconomic health inequalities within the different Spanish healthcare services, using concentration indices. Data were taken from the most recent Encuesta Nacional de Salud (Spanish National Health Survey, 1997) and self-perceived health was used to measure health. It is argued that while inter-regional inequalities in health may have been unaffected by decentralization, intra-regional inequalities may be the undesired consequences of an efficiency-based NHS. The results suggest that devolution may have helped pro-equity policies but only in areas where the private sector is small. PMID- 15799457 TI - Health care cost containment strategies: the Jordanian experience. AB - Jordan's relative success in containing costs is the result of public financing of the health insurance system, the health care system reform strategy, and expanding the primary care network, which allows for cost containment and universal access based on the need for services rather than the ability to pay. The shift of costs from the public to the private sector must be curtailed. The determinants of health care (i.e. environment; human biology; life style; and health care system) are the main factors that determine future spending on health. PMID- 15799459 TI - Complexes and clusters of water relevant to atmospheric chemistry: H2O complexes with oxidants. AB - Experimental observations and data from quantum chemical calculations on complexes between water molecules and small, oxygen-containing inorganic species that play an important role as oxidants in the atmosphere (O(1D), O(3P), O2(X3sigmag), O2(b1sigmag+), O3, HO, HOO, HOOO, and H2O2) are reviewed, with emphasis on their structure, hydrogen bonding, interaction energies, thermodynamic parameters, and infrared spectra. In recent years, weakly bound complexes containing water have increasingly attracted scientific attention. Water in all its phases is a major player in the absorption of solar and terrestrial radiation. Thus, complexes between water and other atmospheric species may have a perceivable influence on the radiative balance and contribute to the greenhouse effect, even though their concentrations are low. In addition, they can play an important role in the chemistry of the Earth's atmosphere, particularly in the oxidation of trace gases. Apart from gas-phase complexes, the interactions of oxidants with ice surfaces have also received considerable advertency lately due to their importance in the chemistry of snow, ice clouds, and ice surfaces (e.g., ice shields in polar regions). In paleoclimate- respectively paleoenvironmental--studies, it is essential to understand the transfer processes from the atmosphere to the ice surface. Consequently, special attention is being paid here to the intercomparison of the properties of binary complexes and the complexes and clusters of more complicated compositions, including oxidants adsorbed on ice surfaces, where ice is considered a kind of large water cluster. Various facts concerning the chemistry of the Earth's atmosphere (concentration profiles and possible influence on radical reactions in the atmosphere) are discussed. PMID- 15799458 TI - The financial burden of malaria in Tanzania: implications for future government policy. AB - Malaria is the cause of more mortality and morbidity in Tanzania than any other disease, in large part due to growing resistance to anti-malarial drugs. This study estimates that over 1% of GDP is devoted to the disease, representing US$2.2 per capita, and 39% of total health expenditure nationally. Government facilities devote almost one-third of their resources to the disease. Private expenditure, primarily on drugs, coils, sprays and bed-nets, represents 71% of total expenditures. Given the dominance of malaria treatment outside Government facilities, strategies to control behaviour in the private sector are critical. Together with regulations on private providers, and other interventions such as promoting the use of bed-nets in rural areas, greater research into and use of information strategies is required. Public policies should be designed to influence behaviour, to encourage households to seek adequate diagnosis of fever and to complete appropriate treatment with the right drugs. PMID- 15799460 TI - Molecular rotation as a tool for exploring specific solute-solvent interactions. AB - Solute-solvent interactions play an important role in determining the physicochemical properties of liquids and solutions. As a consequence, understanding these interactions has been one of the long-standing problems in physical chemistry. This Minireview describes our approach towards attaining this goal, which is to investigate rotational relaxation of a pair of closely related, medium-sized nondipolar solutes in a set of appropriately chosen solvents. Our studies indicate that solute-solvent hydrogen bonding significantly hinders solute rotation. We have also examined the role of solvent size both in the absence and presence of specific interactions and it has been observed that the size of the solvent has a bearing on solute rotation especially in the absence of specific interactions. Our results point to the fact that only strong solute solvent hydrogen bonds have the ability to impede the rotation of the solute molecule because, in such a scenario, hydrogen-bonding dynamics and rotational dynamics transpire on comparable time scales. This aspect has been substantiated by measuring the reorientation times of the chosen solutes in solvents such as ethanol and trifluoroethanol, which have distinct hydrogen-bond donating and accepting abilities, and correlating them with solute-solvent interaction strengths. As an alternative treatment, it has been shown that specific interactions between the solute and the solvent can be modeled as dielectric friction with the extended charge distribution model. This approach is not unrealistic considering the fact that specific as well as non-specific interactions are electrostatic by nature and the differences between them are subtle. PMID- 15799462 TI - Two-component membrane lithography via lipid backfilling. PMID- 15799461 TI - Femtochemistry of inter- and intramolecular hydrogen bonds. PMID- 15799463 TI - Isolated nanoribbons of carbon nanotubes and peapods. PMID- 15799464 TI - A barrier-free nonatomic radical-molecule reaction: CCN + CH4. PMID- 15799465 TI - IR spectra of isolated arenium ions: hydroxyl and halogen substitution effects on aliphatic CH bonds. PMID- 15799466 TI - Infrared spectroscopy of protonated phenylsilane in the gas phase. PMID- 15799467 TI - Direct observation of segregation of sodium and chloride ions at an ice surface. PMID- 15799469 TI - Covalent linking of CdTe nanocrystals to amino-functionalized surfaces. PMID- 15799468 TI - Equilibrium coverage fluctuations: a new approach to quantify reversible adsorption of proteins. PMID- 15799470 TI - What are the implications of nonequilibrium in the O + OH and O + HO2 reactions? AB - Vibrationally excited O2, OH, and HO2 species have been suggested (J. Phys. Chem. A 2004, 108, 758) to provide clues for explaining the "ozone deficit problem" and "HOx dilemma" in the middle atmosphere under conditions of local thermodynamic disequilibrium (LTD), but the question arises of how much LTD will affect the title ozone sink reactions. Besides providing novel kinetic results, it is shown that LTD tends to disfavor ozone depletion relative to traditional atmospheric modeling under Boltzmann equilibration, which is partly due to competition between the various reactive channels. The calculations also suggest that the title LTD processes can be important sources of highly vibrationally excited O2 in the middle atmosphere. Moreover, LTD is shown to offer an explanation for the fact that some down revision of the O + HO2 rate constant, or the ratio of the O + HO2 to O + OH rate constants, is required to improve agreement between the predictions of traditional modeling and observation. This, in turn, provides significant evidence supporting LTD at such altitudes. PMID- 15799471 TI - Pressure effects on Friedel-Crafts alkylation reactions in supercritical difluoromethane. AB - Dielectrometry is used as a novel technique for following the rate of a Friedel Crafts alkylation reaction in supercritical (sc) difluoromethane. Although the process was not optimized, good product yields were obtained and reaction rates were found to be larger than in CO2 at comparable conditions. The reaction order is determined using the initial rate method and the reaction is shown to be first order with respect to both anisole and t-butyl chloride. The reaction rate constant is unaffected by pressures above 120 bar but close to the critical pressure the value decreases with increasing pressure. It is also shown that the product distribution for the alkylation of anisole shows significant pressure dependence with substitution at the ortho-position being favored at lower pressures, which is ascribed to hydrogen bonding. This pressure dependency is not observed in sc CO2 or using toluene as a substrate, which supports the idea that hydrogen bonding may be important in the reaction mechanism. The effect of the different reagents and temperature on the rate of the alkylation reaction was also determined. PMID- 15799472 TI - Testing the pairwise additive potential approximation using DFT: coadsorption of CO and N on Rh (100). AB - The interaction between adsorbates is a key issue in surface science, because these interactions can influence strongly the properties of chemisorbed species with consequences for the thermodynamics and kinetics of surface processes. The simplest representation of adsorbate-adsorbate interactions is based on the assumption that all interactions are pairwise additive. This approach has been satisfactorily used in the modeling of temperature-programmed desorption (TPD) spectra using both continuum and Monte Carlo methods. However, the energies estimated within the pairwise approximation have never been compared to the energies calculated using density functional theory (DFT) methods. We demonstrate that the pairwise additive potential approximation is indeed a good representation of the adsorbate-adsorbate interactions, and that we do not need to include three-body interactions or higher-order terms to estimate the perturbation of the adsorption energy of an adsorbate by the presence of other coadsorbates. Moreover, we show for the first time how DFT can be used to explain the desorption features that one finds in TPD experiments, thus linking the TPD desorption features with actual microscopic configurations. PMID- 15799473 TI - Fabrication of ultrathin films with large third-order nonlinear optical properties. AB - An ultrathin composite film containing anionic Ag-His complexes (His: L Histidine) and oppositely charged BH-PPV was fabricated by means of the electrostatic layer-by-layer self-assembly technique. UV/Vis spectra showed a continuous deposition process of Ag-His complexes and BH-PPV. The film structure was characterized by using small-angle X-ray diffraction, AFM, and SEM. The nonlinear optical properties of the ultrathin film were studied by using the Z scan technique with a laser duration of 8 ns at a wavelength of 532 nm. The film sample exhibited a strong nonlinear saturated absorption, with an alpha2 value of -3.9 x 10(-5) mW(-1) and a self-defocusing effect with an n2 value of -4.78 x 10 (-12) m2W(-1). PMID- 15799474 TI - Focal-point conformational analysis of ethanol, propanol, and isopropanol. AB - Conformational analysis of three small alcohols--ethanol, propanol, and isopropanol--was carried out by systematically improving the basis set and the level of electron correlation. Correlation energy contributions to conformational energies are strongly basis-set-dependent but accurate energy contributions can be obtained by extrapolation to the basis-set limit. At the basis-set limit, second- and third-order electron correlation effects play a significant role for rotations around the CC-OH, HC-CO, and CC-CO bonds. Specifically, second- and third-order correlation effects strongly stabilize structures in which the hydroxylic hydrogen eclipses with the adjacent carbon; a lesser stabilization is present in structures where the CC-OH moiety is in the gauche form. Fourth-order correlation effects to the CC-OH rotation are small due to a partial cancellation of the singles, doubles, and quadruples contribution by the triples contribution. Electron correlation significantly lowers barriers for methyl-group rotations in ethanol and isopropanol, and in these cases the fourth-order correlation effects are noticeable. The relatively large overall importance of third-order correlation energy contributions raises a concern that the inability to accurately estimate this slowly converging contribution may become a limiting factor when highly accurate conformational energies in larger molecules are sought. PMID- 15799475 TI - C-H...O hydrogen bonds in cyclohexenone reveal the spectroscopic behavior of Csp3 H and Csp2-H donors. AB - C-H...O hydrogen bonds in liquid 2-cyclohexen-1-one are studied to assess the vibrational spectroscopic behavior of the Csp2-H and Csp3-H donors. The presence of a pseudo-isosbestic point in the vC = O region supports the assignment of the two observed bands to two species in equilibrium, considered to be the free and 1:1 associated forms. The values of deltaH degrees =-18.5 +/- 0.6 kJmol(-1) and deltaS degrees = -76 +/- 2 J K(-1) mol(-1) for the dimerization through C-H...O hydrogen bonds were obtained from the dimerization constant at different temperatures. The concentration-dependent intensity of the vCH2 band profile is ascribed to the presence of a blue-shifted band from the hydrogen-bonded Csp3-H group. However, the most surprising result is the absence of concentration- or temperature-dependent intensities in the bands assigned to the stretching modes of the Csp2-H donors. PMID- 15799476 TI - Theoretical study of the electronic excited states of tetracyanoethylene and its radical anion. AB - The low-lying electronic states of tetracyanoethylene (TCNE) and its radical anion were studied using multiconfigurational second-order perturbation theory (CASPT2) and extended atomic natural orbital (ANO) basis sets. The results obtained yield a full interpretation of the electronic absorption spectra, explain the spectral changes undergone upon reduction, give support to the occurrence of a bound excited state for the anionic species, and provide valuable information for the rationalization of the experimental data obtained with electron transmission spectroscopy. PMID- 15799477 TI - Through versus cross electron delocalization in polytriacetylene oligomers: a computational analysis. AB - Herein, we report a systematic theoretical investigation of the molecular and electronic properties of unsubstituted polytriacetylene (PTA) and iso polytriacetylene (iso-PTA) oligomers, which are characterized by through and cross pi-conjugation pathways, respectively. The goal of the study is to compare through versus cross conjugation on the basis of the computed molecular geometries of the neutral, anionic, and cationic species, the electron affinities, ionization potentials, excitation energies, and nonlinear optical properties for oligomers up to the nonamer. Differences in the effective conjugation length are directly related to electron delocalization in cross- and through-conjugated pathways. As in the through-conjugated oligomers, that is, the PTAs, the frontier orbitals of the iso-PTA oligomers are delocalized along the entire carbon backbone, suggesting that pi-delocalization can extend through cross-linked carbon atoms. However, in contrast to the PTA oligomers, the bond lengths remain strictly constant and the reduction of the energy gap beyond the trimer is completely due to the correlation contribution. On the other hand, in the anions and cations, the bond lengths do change significantly with increasing chain length. Therefore, oxidation or reduction of the iso-PTA oligomer appears to switch on delocalization through cross-linked carbon atoms. Obviously, the effective conjugation length is specific and depends on the observable considered. PMID- 15799478 TI - Crystallographic and electrochemical characteristics of La(0.7)Mg(0.3)Ni(5.0 x)(Al(0.5)Mo(0.5)x hydrogen-storage alloys. AB - The structure, hydrogen-storage property and electrochemical characteristics of La(0.7)Mg(0.3)Ni(5.0-x)(Al(0.5)Mo(0.5))x (x = 0-0.8) hydrogen-storage alloys have been studied systematically. X-ray diffraction Rietveld analysis shows that all the alloys consist of an La (La,Mg)2Ni9 phase and an LaNi5 phase. The pressure composition isotherms indicate that the hydrogen-storage capacity first increases and then decreases with increasing x, and the equilibrium pressure decreases with increasing x. Electrochemical measurements show that the maximum discharge capacity and the exchange-current density of the alloy electrodes increase as x increases from 0 to 0.6 and then decrease when x increases further from 0.6 to 0.8. Moreover, the low-temperature dischargeability of the alloy electrodes increases monotonically with increasing x in the alloys. PMID- 15799479 TI - A comparative electrochemical study of diffusion in room temperature ionic liquid solvents versus acetonitrile. AB - Measurements on the diffusion coefficient of the neutral molecule N,N,N',N' tetramethyl-para-phenylenediamine and the radical cation and dication generated by its one- and two-electron oxidation, respectively, are reported over the range 298-348 K in both acetonitrile and four room temperature ionic liquids (RTILs). Data were collected using single and double potential step chronoamperometry at a gold disk electrode of micrometer dimension, and analysed via fitting to the appropriate analytical expression or, where necessary, to simulation. The variation of diffusion coefficient with temperature was found to occur in an Arrhenius-type manner for all combinations of solute and solvent. For a given ionic liquid, the diffusional activation energies of each species were not only closely equivalent to each other, but also to the RTIL's activation energy of viscous flow. In acetonitrile supported with 0.1 M tetrabutylammonium perchlorate, the ratio in diffusion coefficients of the radical cation and dication to the neutral molecule were calculated as 0.89 +/- 0.05 and 0.51 +/- 0.03, respectively. In contrast, amongst the ionic liquids the same ratios were determined to be on average 0.53 +/- 0.04 and 0.33 +/- 0.03. The consequences of this dissimilarity are considered in terms of the modelling of voltammetric data gathered within ionic liquid solvents. PMID- 15799480 TI - Simultaneous AFM manipulation and fluorescence imaging of single DNA strands. AB - We report combined atomic force and far-field fluorescence microscopic experiments which allow the simultaneous atomic force manipulation and optical observation of individual dye-labeled DNA molecules. A detailed understanding of the binding properties of DNA to different transparent surfaces is prerequisite for these investigations. Atomic force spectroscopy and fluorescence microscopy of single DNA strands yielded detailed insight into two different types of DNA binding onto transparent polylysine-coated and silanized glass surfaces. We subsequently demonstrate how the different binding can be exploited to perform two types of nanomanipulation experiments: On polylysine, strong electrostatic interactions over the whole length of the DNA strand enable the writing of micrometer-sized patterns. By contrast, the strong pointwise attachment of DNA to silanized surfaces allows horizontal stretching of single DNA strands to lengths exceeding 1.6 times the contour length of the DNA strand. With this new approach it is possible to directly observe the rupture of the strongly bonded DNA strand. PMID- 15799481 TI - High-spin chloro mononuclear MnIII complexes: a multifrequency high-field EPR study. AB - The isolation, structural characterization, and electronic properties of two six coordinated chloromanganese (III) complexes, [Mn(terpy)(Cl)3] (1) and [Mn(Phterpy)(Cl)3] (2), are reported (terpy = 2,2':6'2"-terpyridine, Phterpy = 4' phenyl-2,2':6',2"-terpyridine). These complexes complement a series of mononuclear azide and fluoride Mn(lll) complexes synthesized with neutral N tridentate ligands, [Mn(L)(X)3] (X = F- or N3 and L = terpy or bpea [N,N-bis(2 pyridylmethyl)-ethylamine)], previously described. Similar to these previous complexes, 1 and 2 exhibit a Jahn-Teller distortion of the octahedron, characteristic of a high-spin Mn(III) complex (S = 2). The analysis of the crystallographic data shows that, in both cases, the manganese ion lies in the center of a distorted octahedron characterized by an elongation along the tetragonal axis. Their electronic properties were investigated by multifrequency EPR (190-475 GHz) performed in the solid state at different temperatures (5-15 K). This study confirms our previous results and further shows that: i) the sign of D is correlated with the nature of the tetragonal distortion; ii) the magnitude of D is not sensitive to the nature of the anions in our series of rhombic complexes, contrary to the porphyrinic systems; iii) the [E/D] values (0.124 for 1 and 0.085 for 2) are smaller compared to those found for the [Mn(L)(X)3] complexes (in the range of 0.146 to 0.234); and iv) the E term increases when the ligand-field strength of the equatorial ligands decreases. PMID- 15799482 TI - Wetting of surfactant solutions by alkanes. AB - Ellipsometry, surface tensiometry, and contact-angle measurement have been used to study the transition between partial wetting and pseudo-partial wetting of surfactant solutions by alkanes. In the partial wetting regime, the air-water surface tension is the same with and without alkane. In the pseudo-partial wetting regime, the air-water surface tension is lowered by the presence of alkane, showing that oil is solubilised into the surfactant monolayer. A discontinuous change in the coefficient of ellipticity with increasing surfactant concentration provides unequivocal evidence for the first-order nature of the wetting transitions. Ellipsometry has been used to explore the generality of wetting transitions of alkanes (dodecane, hexadecane, and squalane) on surfactant solutions [dodecyltrimethylammonium bromide, tetredecyltrimethylammonium bromide, dibucaine hydrochloride, and Aerosol OT (AOT)]. Of the systems studied, only hexadecane on AOT solutions did not show a wetting transition. Excess alkane remains as a lens on the surface of the surfactant solutions at all concentrations, but the contact angle is a minimum at the wetting transition. A semiquantitative model for the variation of the contact angle with surfactant concentration is provided. PMID- 15799483 TI - Researchers abroad. PMID- 15799484 TI - Neonatal care for premature infants. PMID- 15799485 TI - Neonatal care for premature infants. PMID- 15799486 TI - Neonatal care for premature infants. PMID- 15799487 TI - Neonatal care for premature infants. PMID- 15799488 TI - Neonatal care for premature infants. PMID- 15799490 TI - Neonatal care for premature infants. PMID- 15799489 TI - Neonatal care for premature infants. PMID- 15799491 TI - Will new ways of creating stem cells dodge the objections? PMID- 15799492 TI - My injury, your blood. PMID- 15799493 TI - Reaching disclosure. PMID- 15799495 TI - The VIP floors. PMID- 15799494 TI - The end of the Great Bioethics Compromise. PMID- 15799496 TI - Conflicts of interest at the NIH: no easy solution. PMID- 15799497 TI - Genetic privacy, abandonment, and DNA dragnets: Is Fourth Amendment jurisprudence adequate? PMID- 15799499 TI - The Hippocratic underground: civil disobedience and health care reform. PMID- 15799498 TI - Justice and the human development approach to international research. PMID- 15799500 TI - The perfect storm of enhancement. PMID- 15799502 TI - Are you a professional? PMID- 15799501 TI - "Ethnic drugs". PMID- 15799503 TI - Efficacy of preoperative antimicrobial skin preparation solutions on biofilm bacteria. AB - RESEARCH ON THE MEDICAL EFFICACY of topical antimicrobials and antibiotics against infections has focused largely on the effect on free-floating, planktonic bacteria. IN THE PRESENCE OF nonbiological surfaces (eg, catheters, prosthetic devices, biomaterials), however, bacteria form highly complex biofilm systems that resist traditional medical treatment. BACTERIAL PATHOGENS commonly found in chronic infections in both the planktonic and biofilm state were challenged with a variety of commonly used topical antimicrobial formulations. BIOFILM BACTERIA were shown to be more resistant to killing than planktonic bacteria. Antimicrobial skin preparation times were adequate to significantly reduce bacterial populations protected in biofilms. PMID- 15799504 TI - Perioperative nurses and patient outcomes--mortality, complications, and length of stay. AB - THE PURPOSE OF THE STUDY described in this article was to identify the relationship between RN staffing factors in the OR and surgical patient outcomes. THE STUDY ADDRESSED two main questions: whether the level of RN staffing in the OR is related to postoperative complications, mortality, and length of stay (LOS) and whether certification, RN agency use, 24-hour staffing, and the performance of multidisciplinary code drills are related to complications, mortality, and LOS. ACCORDING TO THIS STUDY, selected organizational factors in ORs had a significant influence on patient outcomes. PMID- 15799505 TI - Swedish patients' perceptions of a preoperative skin test. AB - PREOPERATIVE PREPARATION is an important aspect of creating a safe and pleasant hospital experience for surgical patients. Evaluating patient satisfaction, however, also depends on determining patients' perceptions of the care they receive. THIS STUDY used a descriptive, qualitative method with a phenomenographic approach to determine how patients perceived undergoing a preoperative skin test that was developed to ascertain what substances might result in intraoperative reactions in patients with allergies, asthma, or eczema. OVERALL, the patients had positive perceptions of the skin test; however, the results indicate a need for improved and individualized preoperative instruction that focuses on patients from a holistic perspective. PMID- 15799506 TI - A critical shortage of surgical technologists creates collaboration between rivals. AB - TWO RIVAL HEALTH CARE SYSTEMS collaborated to meet an increasing demand for surgical technologists in the Milwaukee area by developing and implementing a nine-month surgical technologist program. THIS UNIQUE APPROACH included using grant funds from the Private Industry Council of Milwaukee County to help fulfill local health care organizations' commitment to employee development and adequate staffing. TWENTY-ONE SURGICAL TECHNOLOGISTS graduated from the program and successfully began health care careers; three of the graduates are continuing their education in nursing school. PMID- 15799507 TI - A look at a successful perioperative nurse extern-intern program. AB - IN 2000, St Mary-Corwin Medical Center, Pueblo, Colo, launched a perioperative nurse extern-intern program to address the critical nursing shortage the facility's OR was experiencing. THE PROGRAM HAS THREE LEVELS, all of which focus on applying lessons learned in the classroom to the clinical setting. The final level of the program is designed to help participants' transition from being students to being clinical practitioners. ALL STUDENTS who participated in the program passed the National Council Licensure Examination on the first attempt. In addition, the program has improved employee recruitment and retention rates, reduced orientation and training costs, and resulted in positive feedback from participants. PMID- 15799508 TI - Creating an optimal operating room schedule. AB - SCHEDULING IN THE OR suite is a particularly daunting task for many surgical services managers. CREATING AN OPTIMAL OR schedule requires looking at constraining factors as well as historical data. THIS ARTICLE presents one solution to creating an OR schedule that results in increased use of the OR suite and better profitability. PMID- 15799509 TI - AORN Foundation shares results of scholarship recipient survey. AB - SINCE 1992, the AORN Foundation has been awarding scholarships to provide AORN members with funding to attain their education goals. THE SCHOLARSHIP PROGRAM addresses the daunting challenges facing perioperative nurses, including the perioperative nursing shortage, the aging perioperative workforce, lack of tuition funding from hospitals, and the need for perioperative nurses with a bachelor's or higher degree. IN THE SPRING OF 2004, the AORN Scholarship Committee surveyed scholarship recipients to assess the program's effectiveness in meeting these challenges. THIS ARTICLE provides information about the scholarship program, summarizes the survey results, and discusses the effectiveness of the scholarship program. PMID- 15799510 TI - Graduates of foreign nursing schools in US ORs. PMID- 15799511 TI - Addressing the trauma of anesthesia awareness. PMID- 15799512 TI - Recommended practices for electrosurgery. PMID- 15799513 TI - Recommended practices for endoscopic minimally invasive surgery. PMID- 15799514 TI - AORN guidance statement: sharps injury prevention in the perioperative setting. AB - Occupational exposure to bloodborne pathogens via percutaneous injuries is one of the most serious dangers perioperative team members face on a daily basis. The risk of sustaining a percutaneous injury can be decreased through employee education, clear communication, device engineering, and focused work practice controls. Risk reduction strategies should include specific practices aimed at reducing the unique risks of percutaneous injuries encountered in the perioperative environment. AORN recognizes the various settings in which perioperative RNs practice, and the suggested risk reduction strategies in this guidance statement are intended to be adaptable to any setting where surgical or other invasive procedures are performed. PMID- 15799515 TI - The pros and cons of medical liability reform. PMID- 15799516 TI - Volatile substances from larval habitats mediate species-specific oviposition in Anopheles mosquitoes. AB - Oviposition site selection has been recognized as critical both for the survival and population dynamics of mosquitoes. Volatile substances released from larval habitats have been implicated as potential olfactory cues mediating oviposition. In our continuing studies of cues involved in oviposition site selection, we collected material from the larval habitats of Anopheles albimanus Wiedemann and Anopheles vestitipennis Dyar & Knab, i.e., cyanobacterial mats and Typha domingensis Pers. litter, respectively. The volatile compounds were extracted by freeze-drying the material and trapping the volatilized material on a -55 degrees C titanium condenser. For oviposition trials conducted with wild-caught females, the tested volatile materials were pipetted onto filters floating on the surface of distilled water in Teflon beakers that were placed within oviposition cages. For both species, volatile materials in low concentrations increased oviposition, assessed as egg density, whereas there was a shift to reduced oviposition at higher concentrations. Volatile effect was strongly habitat/species-specific as shown by reciprocal treatment tests. PMID- 15799517 TI - Comparisons of human-landing catches and odor-baited entry traps for sampling malaria vectors in Senegal. AB - A comparative study of human-landing catches (HLCs) and odor-baited entry traps (OBETs) for sampling malaria vectors was conducted in two different bioclimatic areas of Senegal, the Sahelian and Sudano-Guinean phytogeographic zones, from September to December 2002. Mosquitoes were collected by the two methods both indoors and outdoors. The reliability of OBET samples was tested by comparing the two methods. Overall, HLC was more effective indoors and for surveying the anopheline fauna. Both methods were effective in sampling the four known malaria vectors in Senegal [Anopheles gambiae s.s., An. arabiensis Patton, An. funestus Giles, and An. nili (Theobald) ], and mosquito age structures and infectivity rates did not differ between methods. PMID- 15799518 TI - Selection of D2S3, an Aedes aegypti (Diptera: Culicidae) strain with high oral susceptibility to Dengue 2 virus and D2MEB, a strain with a midgut barrier to Dengue 2 escape. AB - Family-based phenotypic selection was used to breed two genetic strains of Aedes aegypti L. that differ in susceptibility to infection with dengue serotype 2 virus (DEN-2) strain JAM1409. A Dengue 2 Susceptible on 3 chromosomes (D2S3) strain was bred from Ae. aegypti aegypti and Ae. aegyptiformosus P1 parents to have a high midgut infection rate (MIR) and a high disseminated infection rate (DIR). A Dengue 2 Midgut Escape Barrier (D2MEB) strain was bred from D2S3 and Houston P1 parents to have a high MIR and a low DIR. After selection in the F2 generation, single strand conformation polymorphism genotypes were determined at cDNA marker loci throughout the genome to test for Mendelian ratios and thereby identify regions containing deleterious or lethal alleles. Both strains were orally challenged with two other DEN-2 genotypes, two DEN-1 genotypes, one DEN-3 genotype, and two DEN-4 genotypes. There were significant differences in MIR and DIR for the different virus strains in both the D2S3 and the D2MEB mosquito lines. PMID- 15799520 TI - Ectoparasites associated to two species of Corynorhinus (Chiroptera: Vespertilionidae) from the Guanacevi mining region, Durango, Mexico. AB - As a part of an inventory of bats in abandoned mines at the municipality of Guanacevi, Durango, Mexico, a sample of long-eared bats (genus Corynorhinus) was collected and ectoparasites were taken. Twenty-three specimens of Corynorhinus mexicanus Allen, 1916, and 18 of Corynorhinus townsendii (Cooper, 1937) were collected in four sampling periods coincident with the seasons. In total, 98 ectoparasites of 10 species and seven families were examined. Five species are recorded for the first time on C. mexicanus and four on C. townsendii. Macronyssus cyclaspis and Trichobius corynorhini had the highest frequency of infestation in both bats. Differences in number of arthropods per bat among seasons were nonsignificant for both species. PMID- 15799519 TI - Using in situ hybridization to detect endosymbiont Wolbachia in dissected tissues of mosquito host. AB - The endosymbiont Wolbachia, extensively occurring in arthropods, usually causes reproductive distortions of the host, such as mosquitoes. In past years, detection of Wolbachia in host tissues has highly relied on transmission electron microscopy (TEM) that is tedious and usually unable to gain satisfactory results without experienced techniques and expensive instruments. Polymerase chain reaction (PCR) recently has become popular in Wolbachia identification. However, necessity of DNA extraction from host individuals or dissected tissues has limited its application in extensiveness and versatility. At present, in situ hybridization has increased its role in examination of various microbes. This report provides a technique for rapid detection and localization of Wolbachia in tissues dissected from mosquitoes and possibly other infected organisms. To detect Wolbachia and to localize them in host tissues more precisely, in situ hybridization by using digoxigenin (DIG)-labeled probes was invented and applied to Wolbachia detection in this study. The results showed that Wolbachia preferentially aggregate in ovarioles, which is consistent with previous observations by TEM. The endobacteria also were detected in salivary glands, mostly in lateral lobes. Ultrastructurally, Wolbachia has been shown to occur in the cytoplasma of salivary gland cells. PMID- 15799521 TI - Insect contribution to horizontal transmission of Reticuloendotheliosis virus. AB - The involvement of insects in Reticuloendotheliosis virus (REV) transmission was examined by testing insects trapped at commercial farms and by controlled feeding experiments using mosquitoes, Culex pipiens L. and house flies, Musca domestica L. We established sensitive methods of REV detection, including reverse transcription-polymerase chain reaction (PCR) for REV-LTR and REV-gag genes, REV antigenemia measurements by enzyme-linked immunosorbent assay, and virus isolation in tissue cultures. A variety of blood-sucking species of insects were trapped at farms with infected poultry and tested, but none were positive. To rule out the possibility of PCR inhibition by insect RNA, spiking experiments were conducted and no interference was observed. Because Cx. pipiens mosquitoes were trapped frequently at farms, we performed feeding experiments with mosquito females fed on a REV-containing tissue culture medium and chicken blood mixture. Virus was detected in the mosquitoes up to 5 h postfeeding, compared with 96 h in the feeding mixture, indicating that Cx. pipiens can only harbor REV for a short period. House flies were suspected to be involved in the virus transmission because they frequently were trapped on positive farms. In contrast to mosquitoes, REV was harbored within the house fly digestive tract for up to 72 h and could infect chickens, as demonstrated by seroconversion and by detection of viral gag-sequence in the cloaca. The current study is supportive for the role of house flies as a mechanical vector of REV among poultry. PMID- 15799522 TI - Drought-induced amplification and epidemic transmission of West Nile virus in southern Florida. AB - We show that the spatial-temporal variability of human West Nile (WN) cases and the transmission of West Nile virus (WNV) to sentinel chickens are associated with the spatial-temporal variability of drought and wetting in southern Florida. Land surface wetness conditions at 52 sites in 31 counties in southern Florida for 2001-2003 were simulated and compared with the occurrence of human WN cases and the transmission of WNV to sentinel chickens within these counties. Both WNV transmission to sentinel chickens and the occurrence of human WN cases were associated with drought 2-6 mo prior and land surface wetting 0.5-1.5 mo prior. These dynamics are similar to the amplification and transmission patterns found in southern Florida for the closely related St. Louis encephalitis virus. Drought brings avian hosts and vector mosquitoes into close contact and facilitates the epizootic cycling and amplification of the arboviruses within these populations. Southern Florida has not recorded a severe, widespread drought since the introduction of WNV into the state in 2001. Our results indicate that widespread drought in the spring followed by wetting during summer greatly increase the probability of a WNV epidemic in southern Florida. PMID- 15799523 TI - Isolation of a myoinhibitory peptide from Leishmania major (Kinetoplastida: Trypanosomatidae) and its function in the vector sand fly Phlebotomus papatasi (Diptera: Psychodidae). AB - Protozoan parasites in the genus Leishmania are ingested by sand flies with blood and multiply in the gut until they are transmitted to a vertebrate host when the sand fly blood feeds again. Infections of the enzootic vector Phlebotomus papatasi Scopoli result in distended midguts with no spontaneous gut contractions. Using a P. papatasi hindgut contraction bioassay, a paralytic factor sensitive to trypsin, chymotrypsin, proteinase-K, and heating at 56 degrees C was detected in crude lysates of Leishmania major promastigotes. Application of parasite lysate to isolated hindguts resulted in reversible, dose dependent inhibition of spontaneous contractions. Mean volume of isolated midguts and hindguts increased by 50-60% after application of L. major lysate. L. major paralytic factor was purified 10(4)-fold over the total protein preparation and yielded a hydrophobic 12-kDa peptide. Myoinhibitory activity eluted as a single peak in reverse phase-high-pressure liquid chromatography. Tandem mass spectrometry resulted in 15 amino acid sequences, three of them sharing 45-73% homology with short hypothetical gene products of undefined function from Pseudomonas, Halobacterium, and Drosophila. This unique protozoan peptide mimics the function of endogenous insect neuropeptides that control visceral muscle contractions. By this novel mechanism, parasites persist in the expanded, relaxed midgut after blood meal and peritrophic matrix digestion. This allows time for development and migration of infective forms, facilitating sand fly vector competence and parasite transmission. PMID- 15799524 TI - Variation among geographically separated populations of Dermacentor andersoni (Acari: Ixodidae) in midgut susceptibility to Anaplasma marginale (Rickettsiales: Anaplasmataceae). AB - Anaplasma marginale is a tick-borne rickettsial pathogen of cattle that is endemic throughout large areas of the United States. Cattle that survive acute infection become life-long persistently infected carriers. In the intermountain west the Rocky Mountain wood tick, Dermacentor andersoni Stiles, is the most common vector of A. marginale. Male D. andersoni acquire A. marginale when feeding on persistently infected cattle and biologically transmit it when they transfer from infected to susceptible hosts. Host-seeking adult D. andersoni were collected from four widely separated natural populations and tested for susceptibility to midgut colonization with A. marginale. Male ticks were fed on calves persistently infected with a strain of A. marginale naturally transmitted by D. andersoni. Gut infection rates ranged from 12.5% of ticks collected from a mountain site near Hamilton, MT, to 62.5% of ticks from a rangeland site near Riley, OR. Sites near Miles City, MT, and Kamloops, British Columbia, Canada, had intermediate levels of susceptibility. The infection rates differed significantly among populations, and the same populations sampled in two consecutive years were not significantly different from one year to the next. Although there was variation among the populations in the size of ticks, size was unrelated to acquisition of gut infection. Quantitative polymerase chain reaction (PCR) demonstrated that there was no significant difference between populations in the mean number of genome copies in the guts of infected ticks. A. marginale from infected ticks was genotyped to confirm that they were all infected with the laboratory strain, and a sample of 682 field-collected D. andersoni was surveyed for A. marginale by nested PCR; none were found to be naturally infected. Infection of the gut is an essential constituent of vector competence for A. marginale; in this study, we have demonstrated significant variation among populations in this key component of vector competence. PMID- 15799525 TI - Detection of Rickettsia felis in a New World flea species, Anomiopsyllus nudata (Siphonaptera: Ctenophthalmidae). AB - The flea and rodent samples studied in this project were collected from field study sites in New Mexico from winter 1998 to spring 2001. During this period, 155 small rodents (14 different species) were live-trapped and combed for the presence of fleas. A total of 253 fleas were collected, comprising 21 species. Two of the 253 fleas collected were polymerase chain reaction (PCR) positive for the Rickettsia 17-kDa protein gene. These two fleas were both Anomiopsyllus nudata Baker, each collected from an individual Neotoma albigula Hartley, on two occasions. Individual fleas positive for the Rickettsia 17-kDa protein gene were then tested with primers targeting the rickettsial genes for citrate synthase (gltA) and two major outer membrane proteins (ompA and ompB). The nucleotide sequences of the PCR products of these two fleas were identical to each other and were 100% (394/394), 100% (1150/1150), 99.8% (469/470), and 99.3% (818/824) similar to the corresponding sequences of the 17-kDa, gltA, ompA, and ompB genes of Rickettsia felis, respectively. Flea homogenates of individual PCR-positive fleas were inoculated into shell vials seeded with Vero cells, and the Gimenez stain technique was used to demonstrate the presence of Rickettsia-like organisms in detached cells found in aspirated medium 19 d after inoculation. These cells were harvested and tested by PCR, targeting portions of the 17-kDa and gltA genes, resulting in products 100% identical to R. felis. This work comprises the first report of R. felis detection in a flea species (A. nudata) endemic to the New World. PMID- 15799526 TI - Borrelia burgdorferi sensu lato infection in larval Ixodes ricinus (Acari: Ixodidae) feeding on blackbirds in northwestern Italy. AB - Birds belonging to 59 species (n = 1,206) were live captured in Piemonte, northwestern Italy, in 2001. Ixodes ricinus (L.) larvae were collected from 59 birds belonging to nine species, and nymphs were recovered on 79 birds belonging to 10 species. Eurasian blackbirds, Turdus merula L., had significantly higher levels of infestation by ticks than other passerine species. Larval I. ricinus of blackbirds peaked in summer, when prevalence was 39% (95% confidence interval 24.2-55.5) and mean number of ticks per host was 3.3 (1.6-7.2), whereas nymphs peaked in spring, when prevalence was 72.2% (54.8-85.8) and mean number of ticks per host was 6.9 (4.4-10.7). Immature I. ricinus were coincidentally aggregated on blackbirds, with 15 blackbirds feeding 67.4% of nymphs and 40.3% of larvae, and coinfestation by both stages was relatively high in summer: Kappa = 0.64 (0.40-0.88). Borrelia burgdorferi sensu lato was identified by polymerase chain reaction (PCR) in 58.3% (35.9-78.5) of larvae with engorgement ratio > or = 3 that were collected from blackbirds. Larvae that were collected from other passerine species gave negative PCR results. Sixteen of 21 PCR-positive samples belonged to B. garinii (76.2%), and five (23.8%) were Borrelia valaisiana. Results of this study suggest that blackbirds play an important role as hosts for immature I. ricinus and as reservoir of Borrelia garinii in northwestern Italy. PMID- 15799527 TI - Observations on distribution and seasonal activity of the Gulf Coast tick in Mississippi. AB - Medical importance of the Gulf Coast tick, Amblyomma maculatum Koch, has been highlighted with the recent discovery of a new rickettsial pathogen associated with this tick. Accordingly, distribution and seasonal collection records of A. maculatum were assessed in Mississippi by using three sources of data: sampling of experimental field plots, national tick collection records, and Mississippi Department of Health human tick biting records. A. maculatum was collected in 17/82 (21%) Mississippi counties, mostly in central and southern regions. Estimates of seasonal activity were made using collection data for 2,217 adult, 11 nymphal, and 426 larval specimens. Adult A. maculatum were collected during March through November, with a peak during late July through early August. Nymphs were collected during February through August, and larvae were collected once during each of June, September, and October. Small sample sizes of the immature stages precluded determination of their peak activity. PMID- 15799528 TI - Entomological characteristics of malaria transmission in Manhica, a rural area in southern Mozambique. AB - From October 1997 to September 1998, an entomological survey was carried out in Manhica, Mozambique, to describe the anopheline population and intensity of malaria transmission. Ten different huts were randomly selected for entomological surveillance throughout the year. CDC light trap collections were conducted during three nights each month. Additional knockdown spraying catches were carried out in the morning, after the last catch. A total of 17,245 Culicinae and 1,251 Anophelinae were collected during the study. There was substantial house to house variation and seasonality in the distribution of Anophelinae population, with a peak in April towards the end of the warm and rainy season. Four species of genus Anopheles (Diptera: Culicidae) were described: Anopheles funestus Giles, Anopheles tenebrosus Donitz, Anopheles arabiensis Patton, and Anopheles merus Donitz. An. funestus constitutes 72.3% of the anopheline population. The estimated sporozoite rate was 1.2% and the average entomological inoculation rate for the area was 15 infective bites per person per year. PMID- 15799529 TI - Cordylobia anthropophaga (Diptera: Calliphoridae) outside Africa: a case of furuncular myiasis in a child returning from Congo. AB - International travel to tropical countries accounts for an increasing incidence of imported diseases. An unusual case of furuncular myiasis due to Cordylobia anthropophaga (Blanchard) is reported in northern France in a 9-mo-old infant, after a 4-mo stay in Congo. A review has been made of the major cases of imported furuncular myiasis due to Cordylobia, as well as identification of second larval instars and management of the myiasis. PMID- 15799530 TI - Insect succession and decomposition of wildlife carcasses during fall and winter in Louisiana. AB - Fourteen fresh animal carcasses were monitored throughout decomposition in a mixed flatwood forest in East Baton Rouge Parish, LA, from 5 October to 7 December 1999 (fall) and from 18 January to 30 March 2000 (winter). Species composition and residency patterns of necrophilous insects were documented for four animal species per seasonal experiment: one Louisiana black bear (threatened species), two white-tailed deer, two alligators, and two swine (experimental reference). Results suggested variation in species composition associated with temperature (fall versus winter conditions) and carcass type. In total, 89 species from 39 families and three classes were manually collected from the seven fall carcasses. Ninety-five species from 38 families and three classes were collected at the seven winter carcasses. Overall arthropod diversity was greatest for fall deer and winter swine carrion. Fall alligator carcasses were associated with fewer taxa than the three mammal species during both seasons. The hairy maggot blow fly, Chrysomya rufifacies (Macquart), was the dominant species of the fall study, impacting developmental rates and overall carrion community structure. The winter study was characterized by prolonged carcass decomposition and reduced insect activity due to fluctuating ambient temperatures. PMID- 15799531 TI - Isolation of Ganjam virus from ticks collected off domestic animals around Pune, Maharashtra, India. AB - Studies on viruses of zoonotic importance in certain villages around Pune were undertaken between December 2000 and January 2002. A total of 1,138 adult ticks belonging to six different species were collected off domestic animals and processed for virus isolation. Six virus isolates were obtained. All six isolates were identified as Ganjam virus by Quick Complement Fixation test and reverse transcriptase-polymerase chain reaction using RNA nucleocapsid gene amplification. Five isolates were from the pools of adult Hemaphysalis intermedia ticks, and one isolate was from a pool of adult Rhipecephalus hemaphysaloides. This is the first report of isolation of Ganjam virus from Maharashtra state of India. PMID- 15799533 TI - Tarasoff at thirty: victim's knowledge shrinks the psychotherapist's duty to warn and protect. PMID- 15799532 TI - Comparison of in vitro and in vivo ectoparasiticide activity of experimental benzimidazole-carbamate with permethrin and amitraz. AB - A series of in vitro and in vivo bioassays were conducted to assess the ectoparasiticide activity of isopropyl-4-nitro-2,6-bis(trifluoromethyl)-1 benzimidazole-carbamate, an experimental benzimidazole-carbamate class compound. This compound was less potent than permethrin against ectoparasiticide susceptible larvae of the lone star tick, Amblyomma americanum (L.) (Acari: Ixodidae); larvae of the southern cattle tick, Boophilus microplus (Canestrini); and adult stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae) in vitro, but it was significantly more potent than permethrin against the Santa Luiza strain of B. microplus known to possess high-level resistance to amitraz and pyrethroids. In contrast, the benzimidazole-carbamate was substantially more efficacious than permethrin when applied topically onto rats that were infested with A. americanum nymphs. These results suggest that this experimental compound may be a viable candidate ectoparasiticide that retains significant activity against resistant B. microplus and also suggests that the benzimidazole-carbamate chemistry may be useful for addressing the growing problem of resistance in ectoparasites. PMID- 15799535 TI - Working towards a better understanding of attention deficit hyperactivity disorder as a legal disability in employment law. PMID- 15799534 TI - Pregnant drug abusers are treated like criminals or not treated at all: a third option proposed. PMID- 15799536 TI - The struggle for rural pharmacies: will Medicare's new privately insured prescription drug covera ge jeopardize valuable pharmacy services for rural seniors? PMID- 15799537 TI - What defines a public health emergency? An analysis of the strategic national stockpile and the National Childhood Vaccine Injury Act: the need for prevention of nonterror national medical emergencies. PMID- 15799538 TI - Effect of alginate composition and purity on alginate microspheres. AB - BACKGROUND: Alginate is commonly used to microencapsulate islets in experiments with islet allografts and xenografts for the treatment of Type I diabetes. The purpose of the present study is to determine the effects of alginate composition and purity on the morphology and size of microspheres. METHODS: Microcapsules produced with the impure alginate types, medium-viscosity high-guluronic acid (IMVG), low-viscosity high-G (ILVG), low-viscosity high-mannuronic acid (ILVM) and medium-viscosity high-M (IMVM) were compared with one another and others generated with a highly purified LVM (HPLVM) alginate. Droplets of 1.5% alginate from an air-syringe pump were gelled in 1.1% CaCl2 solution. While leaving the alginate pressure and needle recess constant, the air-jacket pressure was varied between 9.5-10.5 PPSI to enhance stable microcapsule generation and different batches of microbeads were made from each alginate type. RESULTS: The sizes of the high-guluronic acid alginate microbeads were consistently bigger than those of the corresponding high-mannuronic acid alginate beads at all air-jacket settings. At the optimal air-jacket pressure of 9.0 PPSI, the mean+SD diameter of the IMVG microbeads was 780 + 20 microm, while that of IMVM was 607 + 44 microm (p < 0.0001, n=30). Similarly, the mean ILVG microbead diameter was 816+28 microm compared to 656+26 microm for ILVM capsules (p<0.0001, n=30). Less polymorphism was found with the HPLVM microspheres than with the ILVM microbeads. CONCLUSION: Highly purified high-mannuronic acid alginate will provide smaller, spherical microcapsules suitable for islet cell transplantation. PMID- 15799539 TI - Alginate microparticles for Polymyxin B Peyer's patches uptake: microparticles for antibiotic oral administration. AB - Microparticles with size less than 3 microm, able to be taken up by M cell of Peyer's patches for the drug delivery to the Gut Associated Limphoid Tissue (GALT), were developed in order to improve oral bioavailability of Polymyxin B (PMB). Less than 3 microm alginate microparticles resistant to gastro-intestinal media were prepared by spray-drying technique and cross-linking by calcium ions and chitosan. The cross-linked microparticles were evaluated for PMB content by spectrophotometric method, alginate/PMB interaction by rheological study, cross linking degree by EDS analysis and PMB activity by microbiological assay. By modulating the polymer cross-linking degree, cationic PMB interacted on alginate chains leading to a proper PMB loading as well as antibiotic retention in gastric environment and sustained delivery in intestinal fluid. Moreover, the procedure resulted suitable for PMB biological activity preservation. PMID- 15799540 TI - Polycarboxylic acid nanoparticles for ophthalmic drug delivery: an ex vivo evaluation with human cornea. AB - In ophthalmic drug delivery, a major problem is retaining an adequate concentration of a therapeutic agent in the pre-corneal area. Polycarboxylic acid carriers such as polyacrylic acid and polyitaconic acid in sub-colloidal, nanoparticulate hydrogel form have a strong potential for sustained release of a drug in ocular delivery. Formulations have been prepared of brimonidine loaded in polycarboxylic (polyacrylic and polyitaconic) acid nanoparticles for potential ophthalmic delivery. These particles were prepared by a reverse micro-emulsion polymerization technique with sizes in the range of 50 nm. The loading efficiencies of the drug brimonidine in the particles were shown to be between 80 85% for polyacrylic acid nanoparticles and between 65-70% for polyitaconic nanoparticles. The loading efficiency was also found to be pH dependent. In a preliminary biocompatibility test, human corneal epithelial cells incubated with polyacrylic acid nanoparticles were found to retain their viability, whereas polyitaconic acid nanoparticles were found to be toxic. Two-photon laser scanning microscopic studies of the fluorescently labelled polyacrylic acid nanoparticles and human cornea shows that they are adhesive on the corneal surface. The polyacrylic acid nanoparticles demonstrated a controlled release of the opthalmological drug (Brimonidine) through the human cornea as compared to that of the commercial formulation, Alphagan. PMID- 15799541 TI - Formulation and characterization of triclosan sub-micron emulsions and nanocapsules. AB - Triclosan, a non-ionic, broad spectrum anti-microbic agent, has recently demonstrated its effectiveness as an anti-malarial drug by inhibition of the growth of Plasmodium Falciparum. The aim of this work was to formulate suitable triclosan colloidal carriers with the final objective of obtaining a drug delivery system suitable for a potential anti-malarial oral treatment. Two different nanotechnological approaches were experimented with that could be suitable for developing effective triclosan formulations against this established and re-emerging infectious disease. Sub-micron emulsions were prepared by the solvent displacement method, using different oily amounts in order to vary the drug amount entrapped in the formulation. Chitosan-coated nanocapsules were obtained with chitosan hydrochloride at two different viscosity degrees (Cl 113 and Cl 213). All formulations were appropriately characterized by determining drug loading capacity and encapsulation efficiency and measuring particle size and zeta potential. Morphological characterization of the different systems was performed by TEM analysis, whereas release studies were carried out by reverse bag dialysis method. All preparations resulted stable. Cl 113-coated nanocapsules appeared particularly suitable as triclosan carriers for obtaining a systemic drug release, owing to both chitosan's good mucoadhesive and enhancer properties as well as the effectiveness shown by its coating in adequately controlling drug release rate. PMID- 15799542 TI - Novel solvent-based method for preparation of alginate beads with improved roundness and predictable size. AB - Attempts to determine conditions or processes within alginate gel beads often suffer from inaccuracies due to an improper roundness of the analysed beads. Therefore, a novel solvent-based method for the preparation of alginate beads with improved shape was developed: An aqueous solution of 2% (w/v) alginate in water was injected into a solvent layering consisting of hexane, n-butanol, n butanol with 1% (w/v) CaCl2 and finally 2% (w/v) CaCl2 in water. Beads of up to 3.5 mm in diameter obtained with this method had a roundness which was approximately 5% better than comparable beads prepared by dropping an alginate solution into a CaCl2-hardening bath. This was determined by a software supported quantitative analysis of bead size and shape. Additionally, the novel solvent based method allows for highly reproducible preparation of alginate beads with exactly predictable sizes. The biggest beads obtained with this method were 9 mm in diameter. Thus, with the solvent-based preparation of alginate beads it is now possible to easily obtain beads of exactly the type needed for a specific analytical purpose. PMID- 15799543 TI - The mechanism of surface-indented protein-loaded PLGA microparticle formation: the effects of salt (NaCl) on the solidification process. AB - The purpose of this study was to evaluate ovalbumin (OVA) leakage pathways and to explore the mechanism of the surface-indented microparticle formation in the preparation of OVA-loaded microparticles. OVA-loaded poly (D,L-lactic-co-glycolic acid) (PLGA) microparticles were prepared by a water-in oil-in water (w/o/w) solvent evaporation method associated with varied NaCl (NaCl) concentrations and adjusted with urea at 1240mOsm kg(-1) in the external aqueous phase. To evaluate dichloromethane (DCM)-related OVA leakage, three stirring rates, 600, 800, 1000rpm at 25 degrees C were carried out during the solvent evaporation stage. Both DCM and OVA levels in the external phase medium and total dispersion were sampled and measured. The time course of particle characteristics was evaluated by microscopy or SEM photography. The surface adsorptive capacities of the prepared microparticles were measured by using bovine serum albumin conjugated with fluorescein isothiocyanate (FITC-BSA). The findings were that the DCM related OVA leakage accounted for approximately 34%, of the total leakage. By combining NaCl in the external phase, a faster solidifying crust-like structure was formed as a barrier to remarkably reduce OVA loss and improve OVA content from 40.1 to 72.8 microg mg(-1). The yield and OVA content for formulations containing NaCl were much improved by the ionic effect, in addition to the osmotic effect. The total entrapment efficiency was also highly increased from 43 to 72%. The formations of the crust-like surface structure of the microparticle were affected by entrapped drugs, salt content in the external phase and aqueous volume in the inner phase. A scheme was proposed to interpret the formation mechanism of the surface-indented microparticles. In comparison to the surface smooth microparticles, the surface adsorptive capacities of the surface-indented microparticles were highly improved from 26.6 to 87.0%, determined by the adsorption of FITC-BSA. PMID- 15799544 TI - Preparation of sodium fluoride-loaded gelatin microspheres, characterization and cariostatic studies. AB - Sodium fluoride-loaded gelatin microspheres (NaF-GMS) were prepared using double phase emulsified condensation polymerization. The average diameter of microspheres was (11.33+/-5.56) microm. The drug content and encapsulation efficiency were 8.80% and 76.73%, respectively. The fluoride releasing profiles of NaF-GMS in physiological saline and artificial saliva (pH 4.5, pH 6.8) showed that NaF-GMS had a sustained-release property and fluoride release rate was increased in pH 4.5 artificial saliva. Experiments conducted in rabbits' oral cavity using NaF-GMS and NaF solution as control revealed NaF-GMS could maintain oral fluoride retention longer than NaF solution. Cariostatic abilities of NaF GMS including demineralization prohibition in vitro, fluoride deposition in artificial dental plaque and the ability of targeting to cariogenic bacteria were investigated in artificial dental plaque. The results indicated NaF-GMS with lower fluoride concentrations could achieve equivalent cariostatic effect to the concentrated NaF solution, at the same time, could prolong fluoride retention in dental plaque. Microscopic observation showed that NaF-GMS carrying fusion protein of glucan-binding domain could adhere more bacteria than NaF-GMS and this might indicate the possibility of targeting to cariogenic bacteria when NaF-GMS were properly modified. PMID- 15799545 TI - Literature alerts. PMID- 15799546 TI - Breaking the news in amyotrophic lateral sclerosis. AB - Diagnosis communication is a mainstay of the patient-physician relationship and in ALS represents the beginning of the treatment. In Europe, the diagnosis is now communicated to most patients, but with some differences from north to south. Communication is the basis of the patient's autonomy. When adequately informed, patients are able to consider their preferences about life-extending interventions. However, this discussion is sometimes entertained too late. Both the style and the context of this discussion have profound impact on patient satisfaction. The content of the communication should include all relevant information about the disease and should be performed honestly while preserving hope. Collusion with relatives can create difficulties in the relationship with a patient and isolate them from their family. Diagnostic communication is an ongoing process and should form part of every follow-up visit. Communication with the dying patient is often reinforced by non-verbal messages, acknowledgements of patients' emotions, and by listening. Written material may increase satisfaction of patients and their relatives, but should never substitute for face-to-face discussion. PMID- 15799547 TI - Sequential designs for clinical trials in amyotrophic lateral sclerosis. AB - BACKGROUND: The objective of this paper is to discuss the sequential trial design and its advantages in clinical trials for ALS. The sequential trial design is an alternative to the classical trial design, which permits stopping a study as soon as a treatment effect can be significantly demonstrated or denied. METHODS: As an example of a sequential survival analysis, a recently completed clinical trial is described. A secondary outcome measure used in the same trial, the decline of the vital lung capacity, was re-analyzed sequentially, in order to illustrate the use of the sequential method for a non-survival variable. To compare the classical with the sequential trial design, the number of patients needed in trials aiming at survival effects ranging from 10% to 20% with a power of 80% or 90% was calculated for both designs. RESULTS: In the given examples the time needed to prove the null hypothesis in the survival analysis, and the number of patients needed to prove the null hypothesis in the analysis of the vital capacity is lower than would have been the case in a classical analysis. In 18 of 24 different situations, the chance is at least 90% that with a sequential trial design fewer patients are needed. CONCLUSIONS: We argue that, particularly for ALS trials, a sequential design may be superior to a classical trial design, as it most often requires fewer patients than classically designed trials of equal power, and more importantly may avoid unnecessary continuation. PMID- 15799548 TI - Survival of 793 patients with amyotrophic lateral sclerosis diagnosed over a 28 year period. AB - We reviewed retrospectively 793 consecutive patients discharged with ALS diagnosis between 1971 and 1998 to identify survival predictors. Vital status in July 2001 was known for all patients. Mean age at onset was 56 (SD 11.7) years and mean age at diagnosis was 57.4 (SD 11.5) years. Onset symptoms were bulbar in 20.8% of patients. The median survival time from symptoms onset was 2.9 years (95% CI 2.7-3.1). Survival from onset was 93% after one, 48% after three and 24% after five years. Median survival was shorter in patients with time between onset and diagnosis <12 months than in those diagnosed> 23 months; (2.1 years vs. 5.9 years; P<0.001). Patients with onset after age 65 had a 4.2 times greater risk of death than patients less than 40 years. Bulbar onset was associated with 1.4 times greater risk of death than spinal onset. Patients diagnosed after 1990 had longer median survival (3.3 years) than those diagnosed in 1971-1979 (2.4 years) (P<0.001). As expected, age, bulbar onset and short time from onset to diagnosis were independent predictors of survival. We also found a significant increase in survival over time, probably due to improved comprehensive treatment of ALS patients in Italy. PMID- 15799549 TI - Inflammation in amyotrophic lateral sclerosis spinal cord and brain is mediated by activated macrophages, mast cells and T cells. AB - Recent studies have shown inflammatory markers in affected neural tissues of amyotrophic lateral sclerosis (ALS) patients. We examined immunocytochemically spinal cord tissues of six patients with ALS, two with corticospinal tract degeneration secondary to cerebral infarcts and three control subjects without neuropathologic abnormalities. ALS spinal cords had dense macrophage infiltration (one log greater than control spinal cords) involving the white and gray matter, with heaviest infiltration of lateral and ventral columns and, in one patient, prefrontal gyrus and the occipital lobes of the brain. Macrophages in ALS spinal cord showed strong expression of cyclooxygenase-2 (COX-2) (one log greater than control tissues) and inducible nitric oxide synthase. In the gray matter, macrophages surrounded and appeared to phagocytize neurons (NeuN-positive) that appeared to be dying. Vessels showed damage to the tight junction protein ZO-1 in relation to perivascular CD40 receptor-positive macrophages and CD40 ligand positive T lymphocytes. ALS spinal cords, but not control cords, were sparsely infiltrated with mast cells. In control cases with corticospinal tract degeneration following hemispheric cerebral infarction, macrophage infiltration of the white matter was COX-2-negative and restricted to lateral and anterior corticospinal tracts. Our data suggest that inflammation in ALS spinal cord and cortex is based on innate immune responses by macrophages and mast cells and adaptive immune responses by T cells. PMID- 15799550 TI - CGP 3466B has no effect on disease course of (G93A) mSOD1 transgenic mice. AB - BACKGROUND: There is an accumulating body of evidence that apoptosis is involved in the motor neuron death that occurs in ALS, and in the (G93A) mSOD1 transgenic mouse model (mSOD1 mice). CGP 3466B, a tricyclic propargylamine structurally related to (-)-deprenyl, was found to inhibit apoptosis in a wide variety of in vitro and in vivo models. We therefore studied the effect of CGP 3466B in mSOD1 mice. METHODS: As the effect of CGP 3466B was previously reported to have a bell shaped curve, we performed a dose-ranging study. High-copy G93A mSOD1 mice were treated subcutaneously from the age of 50 days until death with four concentrations of CGP 3466B (0.39 microg kg(-1), 3.9 microg kg(-1), 39 microg kg( 1), and 390 microg kg(-1)). Behavioural tests were performed daily to determine disease onset, disease progression and survival. At the age of 110 days, two mice per group were sacrificed for histopathological analysis of the lumbar ventral horn and for semiquantitative analysis of motor neuron number. RESULTS: We observed no effect on disease onset, disease progression, or survival of the mice. We also did not observe a significant effect on the number of motor neurons due to CGP 3466B. CONCLUSIONS: We conclude that in high-copy G93A mSOD1 mice, chronic subcutaneous treatment with CGP 3466B offers no clinical benefit. PMID- 15799551 TI - Development and implementation of the Dutch protocol for rehabilitative management in amyotrophic lateral sclerosis. AB - INTRODUCTION: In the Netherlands, rehabilitation care plays an important role in the symptomatic and palliative treatment of ALS patients. However, until 1999 there were no guidelines or practice parameters available for the management of ALS. Therefore, the Dutch protocol for rehabilitative management in ALS was developed. We describe the development process, the outcome and implementation of the protocol. METHODS: A concept management protocol was written and the Delphi method was selected to develop the protocol further. This method comprises repetitive discussion sessions from postulates, using a combination of written questionnaires and work-conferences. Between 80 and 90 persons (rehabilitation team members of different professional backgrounds and neurologists) were involved in this process. The protocol was implemented by sending it to all consultants in rehabilitation medicine in the Netherlands; they were asked to inform all the treatment team members about the final protocol and to implement it in their treatment of ALS patients. RESULTS: The protocol was developed in 1999, implemented in 2000 and evaluated in 2001. Recommendations for improvement were made during the evaluation and improvements are currently being developed by an expert group. The protocol is widely used (88.9%) by consultants in rehabilitation medicine and their treatment teams in the Netherlands. CONCLUSIONS: The Dutch protocol for rehabilitative management was developed to provide an optimal and adequate care plan for patients with ALS. It is widely used in the Netherlands. PMID- 15799552 TI - Physical decline and quality of life in amyotrophic lateral sclerosis. PMID- 15799553 TI - Inter-evaluator reliability of the ALS functional rating scale. AB - The ALS Functional Rating Scale-Revised (ALSFRS-R) is used in the clinic to chart progression and as a primary endpoint measure in clinical trials. In the original description of the scale, evaluations were to be performed by the patient or caregiver, but ratings are commonly performed by health care providers. We determined whether there are differences in scoring based on whether the provider, patient or caregiver performed the evaluation. Overall, all evaluators assessed similar changes in function over time, but significant visit-specific differences were found with higher scores when the patient and provider were the evaluators. We modeled how a change in evaluators at the end of a 9-month trial would influence statistical analysis, if the patient was unable to travel to the study site and the final assessment was performed by telephone. If 25% of the final visit assessments were performed by the patient, in place of the provider, the change in score is less than one point on the ALSFRS-R (-0.7: 95% Confidence Interval -2.1 to 0.3). We conclude that the ALSFRS-R can be successfully used even if evaluators change. PMID- 15799554 TI - Changes in the management of ALS since the publication of the AAN ALS practice parameter 1999. AB - OBJECTIVE: To determine if the publication in 1999 of the AAN Practice Parameter on ALS was associated with an improvement in the standard of management of the disease. METHODS: Data on 646 patients enrolled in the ALS CARE database and on 465 patients who died in the period May 2001 to November 2002 were compared with similar data obtained from the database from 1996 to May 1999. RESULTS: The specialty ALS clinics were the most important source of information about ALS. The internet was a source for 39%. The treatment of sialorrhea, pseudobulbar emotional lability, and failure of swallowing and breathing had all improved significantly in the period after the publication of the Practice Parameter. However many patients still did not receive a gastrostomy tube or non-invasive positive pressure ventilation when indicated by the Practice Parameter, mainly because of lack of patient compliance. Cost was the main reason why 41% of patients did not receive riluzole, though they spent a third of the cost of this medication on alternative medicines. CONCLUSIONS: The publication of the AAN Practice Parameter was associated with improvement in the standard of care. Most cases in the database come from specialized ALS centers, and further information on the community care of ALS patients is needed. PMID- 15799555 TI - Depression and bulbar involvement in amyotrophic lateral sclerosis. AB - OBJECTIVE: Patients with amyotrophic lateral sclerosis (ALS) often develop depressive symptoms. Little is known of the factors that predict or influence depression in ALS patients. PATIENTS AND METHODS: In 41 ALS patients we compared a self-rating depression scale with the ALS Functional Rating Scale (ALS-FRS), duration of disease, age, sex, education and participation in a self-help group. RESULTS: There was no significant relation between the total ALS-FRS score and the self-rating depression scale. In contrast, we found a significant correlation between the swallowing (r=-0.453; P=0.003) and breathing (r=-0.333; P=0.033) items of the ALS-FRS and the depressive scale. Depressive symptoms were negatively correlated with the duration of the disease (r=-0.377; P=0.016); there was no influence of age or sex. CONCLUSION: We found no evidence for a direct association between the loss of physical ability in general and depression, but for a decrease of depressive symptoms in relation to the length of time since diagnosis. Therefore, depressive symptoms in ALS patients seem to occur mainly as a depressive reaction following the communication of the diagnosis. In addition, patients with bulbar and respiratory symptoms should carefully be screened for depressive symptoms. PMID- 15799556 TI - A Phase I safety study of hyperbaric oxygen therapy for amyotrophic lateral sclerosis. AB - BACKGROUND: Vascular endothelial growth factor and mitochondrial abnormalities have been described in ALS and its animal models. We have reported that hyperbaric oxygen (HBO) treatment delayed the onset of weakness in the wobbler mouse. OBJECTIVE: To perform a Phase I safety study of HBO in patients with ALS. METHODS: Five patients with ALS were treated for 60min with 100% oxygen at 2 atmospheres pressure daily for five days a week for four weeks. The patients reported any deterioration in their condition after each treatment, and their neurological condition was measured serially during the four weeks of the treatment, and for four further weeks. RESULTS: Four patients reported decreased fatigue, while one patient dropped out at three weeks because of increased fatigue. Maximum isometric voluntary contraction (MVIC) of all muscle groups except right hand grip improved significantly by up to 97%. Most improvement occurred during the four weeks after treatment. It is possible that the improvement in muscle strength was a placebo or a learning effect, though no such effects have been detected in prior therapeutic trials in ALS using MVIC. No change was detected in other measures of neuromuscular function. CONCLUSIONS: A longer duration, placebo controlled trial in a larger number of patients is needed to determine the safety and efficacy of HBO. Until that is completed, it is not recommended that ALS patients should be treated with HBO. PMID- 15799557 TI - Randomised controlled trial in non-invasive ventilation: what trial? PMID- 15799558 TI - Monitoring of the generation of non-mutagenic benzoic acid compounds during the manufacture of erythrosine. AB - The generation of 2-(2',4'-dihydroxybenzoyl) benzoic acid (DHBBA) and 2-(2',4' dihydroxy-3',5'-diiodobenzoyl) benzoic acid (DHDIBBA) during the manufacture of erythrosine (Food Red No. 3) was examined. DHBBA is formed as an intermediate during the synthesis of fluorescein, and as fluorescein is produced, it is gradually consumed. However, under inappropriate reaction conditions, it remains in the resulting fluorescein at the termination of synthesis. DHDIBBA is easily obtained by the iodination of DHBBA. These compounds are also found when erythrosine is heated in excessive alkali. The results of the Ames test using DHBBA and DHDIBBA showed they did not possess mutagenic activity. The results clearly demonstrated that establishment of an upper limit for DHBBA and DHDIBBA is important in the quality control of fluorescein and erythrosine. PMID- 15799559 TI - Improved enzymatic method for the rapid determination of histamine in wine. AB - Fermented foods are frequently contaminated by histamine generated by microorganisms possessing histidine decarboxylase activity. The ingestion of large amounts of histamine can cause serious toxicological problems in man. Thus, it becomes important to set a reliable method for rapid histamine quantification in foods. The detection of bacteria exhibiting histidine decarboxylase activity is also important to estimate the risk of contamination of food. Previous enzymatic methods used to quantify histamine in fish gave erroneously high values due to interference when applied to wine. A new enzymatic method is described that allows the direct determination of histamine concentrations in this type of sample. It can be used for the detection of histamine in synthetic media, grape must or wine (white, rose, red) without polyphenols or sugar interferences. This new enzymatic method shows a good correlation (R2 = 0.996, p < 0.001) between the histamine concentrations and absorbances in the interval 0.4-160 mg l(-1). Comparison between this enzymatic method and a high-performance liquid chromatography method showed a high correlation (R2 =0.9987, p<0.001). A miniaturized enzymatic method is also proposed, which is particularly useful when high numbers of samples must be analysed. PMID- 15799560 TI - Effects of storage time and temperature on the concentration of ethyl carbamate and its precursors in wine. AB - A 3-year study was carried out on the effects of time and temperature on the concentration of ethyl carbamate in wine. The study monitored the changing concentration of ethyl carbamate and of urea and citrulline, which are two major precursors of ethyl carbamate in wine. In addition to the formation of ethyl carbamate, both urea and citrulline decayed in other reactions. Kinetic analysis was carried out to model the formation of ethyl carbamate and its dependence on the concentrations of ethanol, urea and citrulline. This led to the development of an equation that can be used to predict the concentration of ethyl carbamate in wine at the point of consumption, resulting from any given storage time and temperature profile. The results were in good agreement with data obtained from similar studies. PMID- 15799561 TI - Survey of butyltin compounds in 12 types of foods collected in China. AB - Twelve representative food types including beans, fruit, vegetables, fish, meat, potatoes, sugar, beverages, alcohol, eggs, milk and corn were collected from four districts in China for the analysis of butyltin compounds. The hyphenated system of capillary gas chromatography-flame photometric detector using quartz surface induced tin emission was used to measure proyylated butyltins in various samples. Low levels (ng g(-1)) of butyltin compounds were found in some samples and possible sources of contamination and their significance are discussed. PMID- 15799562 TI - Analysis of heat-processed corn foods for fumonisins and bound fumonisins. AB - Thirty retail samples of heat-processed corn foods, i.e. corn flakes, corn-based breakfast cereals, tortilla chips and corn chips, were analysed for fumonisins- fumonisin B1 (FB1), fumonisin B2 (FB2) and hydrolysed FB1 (HFB1)--as well as for protein- and total-bound FB1. Bound (hidden) fumonisins cannot be detected by conventional analysis. Improved methods for the determination of bound FB1 were developed. The protein-bound FB1 was extracted with 1% sodium dodecylsulfate (SDS) solution. The SDS, which interfered with high-performance liquid chromatography (HPLC) analysis, was then separated from protein-bound FB1 by complexing with methylene blue followed by solvent extraction and hydrolysis with 2 N KOH. To measure total-bound FB1, the sample itself was hydrolysed with KOH. In both cases, clean-up was accomplished on an OASIS polymeric solid-phase extraction column and the bound fumonisins were determined by HPLC measurement of HFB1. Fourteen of 15 samples of corn flakes and other corn-based breakfast cereals analysed contained detectable levels of FB1 with a mean in positive samples of 67ng g(-1) (13-237 ng g(-1)). Two samples also had detectable levels of FB2 (21-23ng g(-1)). Bound FB1 was found in all samples; the mean protein bound FB1 measured was 58 ng g(-1) (22-176 ng g(-1)) and the mean total-bound FB1 measured was 106 ng g(-1) (28-418 ng g(-1)), reported as FB1 equivalents after correction for recoveries of HFB1. There was an average of about 1.3 times more FB1 in the bound form compared with extractable FB1, and this was about twice as much as protein-bound FB1. Seven of the 15 samples of alkali-processed corn-based foods, such as tortilla chips and corn chips, contained FB1 and three contained HFB1 with means in measurable positive samples of 78 (48-134) and 29 (13-47) ng g(-1), respectively. Five of these alkali-processed corn foods contained bound FB1; the mean measurable protein-bound FB1 was 42 ng g(-1) (39-46 ng g(-1)) and the mean measurable total-bound FB1 was 100 ng g(-1) (54-209 ng g(-1)). HFB1 derived from bound FB1 in selected samples was confirmed by HPLC with mass spectrometry (MS). PMID- 15799563 TI - Method of test and survey of caprolactam migration into foods packaged in nylon 6. AB - An analytical method for the determination of the nylon-6 monomer caprolactam in foods is described. The foodstuff was extracted with ethanol: water (1:2) containing capryllactam as internal standard and the extract was defatted using hexane. The extract was analysed by liquid chromatography coupled with mass spectrometry. The test method was calibrated down to 0.7 mg kg(-1). The repeatability of the method was good, with a relative standard deviation of 9% at the 15 mg kg(-1) level. The method was demonstrated to be accurate in an independent external check sample exercise. The new method was applied to the analysis of 50 retail foodstuffs packaged in nylon-6. Caprolactam was detected and confirmed in nine of the 50 food samples, in the range 2.8-13 mg kg(-1). The presence of caprolactam was indicated in a further 15 samples, in the range 0.8 11 mg kg(-1), but these samples did not meet all of the five confirmation criteria applied. All migration levels (both confirmed and unconfirmed) were below the European specific migration limit for caprolactam, which is 15 mg kg( 1). The average migration for all 50 samples, setting non-detectables at half the limit of detection, was 2.6 mg kg(-1) with a standard deviation of 3.1 mg kg(-1) (n = 50). All samples found to contain detectable levels of caprolactam migration were for applications involving heating the food in the packaging. They were packs of, for example, sausage meat for which the food would have been heat processed in the nylon casing, or they were nylon pouches for heating foods by boiling, microwaving or roasting. PMID- 15799564 TI - Partition coefficients in food/packaging systems: a review. AB - Food contamination can result from various interactions between food and packaging materials. Migration of volatiles, additives, monomers and oligomers from packaging materials into food or adsorption of volatile compounds from the food by the polymer are important considerations from safety, hygienic and economic points of view. The term 'migration' includes two phenomena (partition and diffusion) that can be important in determining the concentration of contaminants in a food system at any time. An estimation of the partition coefficient, K, in food/packaging systems has been the major objective of numerous different studies. Various parameters can influence K such as temperature, pH, the chemical structure of the migrant, molecular size and structure, fat content, and degrees of crystallinity. Some theoretical approaches such as the quantitative structure-property relationship method could be of interest in the near future. PMID- 15799565 TI - Feasibility study for the development of certified reference materials for specific migration testing. Part 1: initial migrant concentration and specific migration. AB - The paper describes a project with the main objective of developing the know how to produce certified reference materials (CRMs) for specific migration testing. Certification parameters discussed are the initial concentration of the migrant in the polymer (C(P),0) and the specific migration into a food simulant under certain temperature/time conditions. Sixteen preliminary candidate CRMs were defined and produced. The most important polymers (low- and high-density polyethylene (LDPE and HDPE), polypropylene (PP), polystyrene (PS), polyethylene terephtalate (PET), plasticized polyvinyl chloride (PVC), rigid PVC, polyamides (PA)) and additives as well as monomers representing different physicochemical properties as target substances for migration were chosen. The stability and homogeneity of the migrants in the materials were tested and methods for the determination of the certification parameters were developed and validated. > From the 16 materials produced, the six most suitable CRM candidates (LDPE//Irganox 1076/Irgafos 168, LDPE//1,4-diphenyl-1,3-butadiene (DPBD), HDPE//Chimassorb 81/Uvitex OB, PP homo//Irganox 1076/Irgafos 168, HIPS, 1% mineral oil//styrene, PA 6//caprolactam) were selected. The feasibility of CRM production for the six candidate materials was demonstrated and a trial certification exercise was performed with participation of all four partner laboratories. All six materials showed suitable properties for future production as certified reference materials. PMID- 15799566 TI - Nodal disease in temporal bone squamous carcinoma. PMID- 15799567 TI - Round window membrane and labyrinthine pathological changes: an overview. AB - The round window membrane is considered the most likely pathway from the middle to the inner ear. Various substances placed in the middle ear have been seen to pass through the round window membrane. Once toxic substances or inflammatory mediators such as cytokines and nitric oxide enter the inner ear, various inner ear sequelae such as labyrinthitis, endolymphatic hydrops, sensorineural hearing loss or more insidious diseases can occur. PMID- 15799568 TI - Enhanced sodium absorption in middle ear epithelial cells cultured at air-liquid interface. AB - CONCLUSION: As we demonstrated previously that transcription of alpha-ENaC was correlated with oxygen tension in the culture medium, this study suggests that the increase in alpha-ENaC expression observed under ALI conditions may result from greater oxygenation of ME cells. OBJECTIVE: The physiology of the middle ear (ME) is primarily concerned with keeping the cavities fluid-free, to allow transmission of sound vibrations from the eardrum to the inner ear. ME epithelial cells are thought to play a key role in this process as they actively absorb sodium and water in order to clear any excess fluid present in the cavities. MATERIAL AND METHODS: As an air-liquid interface (ALI) model has been shown to improve differentiation and enhance sodium absorption in other respiratory epithelia, we established an ALI model for ME cells. RESULTS: ME cells cultured under ALI conditions exhibited a fourfold increase in sodium absorption, which was not related to either a metabolic effect or to enhanced morphological differentiation, but instead to an increase in expression of the alpha-subunit of the epithelial sodium channel (alpha-ENaC). PMID- 15799569 TI - Heat shock protein 70 delays gentamicin-induced vestibular hair cell death. AB - In this study, geranylgeranylacetone (GGA) was shown to induce heat shock protein (HSP)70 in the vestibular end organs of the guinea pig and to alleviate gentamicin (GM) ototoxicity. This was accomplished without thermal preconditioning. In isolated guinea pig vestibular end organs we demonstrated possible prophylactic (preventive) effects of GGA on GM ototoxicity by actively inducing HSP70. When HSP70 was pre-incubated with GGA, its content in sensory cell cytoplasm and transitional dark cells was increased. Pre-incubation of vestibular end organs with GGA gave sensory cells partial protection from GM toxicity. These findings show that administration of GGA can protect vestibular sensory cells from GM ototoxicity and suggest that induction of HSP70 by GGA may be a useful adjunct for the treatment of vestibular disorders. PMID- 15799570 TI - Application of a rapid-prototyped temporal bone model for surgical planning. AB - CONCLUSION: This 3D prototyped model constructed using a selective laser sintering method serves as a good material for surgical simulation of a malformed ear. OBJECTIVE: In order to perform surgery of the malformed ear, a thorough understanding of the anatomy of the ear is required, together with sufficient experience. The aim of this study was to investigate the validity of a prototyped temporal bone model for surgical simulation. MATERIAL AND METHODS: Simulated 3D models of two cases of congenital aural atresia were prototyped using a selective laser sintering method. Conventional surgical instruments were used to dissect the models. RESULTS: Abnormal structures of the middle ear, such as the ossicles, labyrinth and facial nerve, were identified by dissecting the model. Presurgical dissection informed the surgeon of the optimum orientation for surgery. The model was disinfected and could be handled by the surgeon in the operating room. Surgery could be carried out safely although in one case it was canceled because of the highly abnormal location of the middle ear cleft and facial nerve. PMID- 15799571 TI - Experimental study of an adjustable-length prosthesis in a temporal bone model. AB - CONCLUSIONS: This prosthesis has the advantage of rapid adjustment at the time of insertion in order to achieve optimal tension and, as a result, optimal sound transmission. OBJECTIVE: To test the acoustic performance of a new, adjustable incus replacement prosthesis in a human temporal bone model. MATERIAL AND METHODS: Experiments were performed in seven human temporal bones, before and after removal of the incus and insertion of the prosthesis. The input comprised 406 pure tones ranging in frequency between 0.1 and 10 kHz at an intensity of 80 dB SPL at the tympanic membrane. The output measurement was stapes footplate displacement, determined by means of a laser Doppler vibrometer. Three lengths of the prosthesis were investigated: optimal, optimal +0.2 mm and optimal +0.4 mm. RESULTS: The optimal-length prosthesis produced similar results to those of an intact middle ear. The slightly longer prostheses decreased middle ear sound transmission at all test frequencies, except those near 1.5 kHz. PMID- 15799572 TI - Biocompatibility of vitallium as ossicular reconstruction material in the middle ear: experimental animal study. AB - CONCLUSION: Although long-term data will be necessary for confirmation, the result of this preliminary study indicates that vitallium may be a good alternative material for ossicular replacement prostheses in the middle ear. OBJECTIVES: To investigate the biocompatibility of vitallium (Co-Cr-Mo) as ossicular reconstruction material in the rabbit middle ear, and to compare the results with those obtained with titanium, well known as a highly biocompatible material, and non-implanted control groups. MATERIAL AND METHODS: Eighteen female New Zealand White rabbits were anesthetized. The tympanomeatal flap was elevated and 12 vitallium and 12 titanium implants were placed in the bulla away from the ossicles in 24 middle ears. Six rabbits were used as non-implanted controls. All animals were sacrificed under general anesthesia on the 180th day after implantation. The temporal bones were removed, fixed in 10% buffered paraformaldehyde and decalcified for a week in EDTA. Tissue samples were then prepared using an Autotechnicon and embedded in paraffin. Sections (30-microm thick) were cut with a microtome, stained with hematoxylin-eosin, von Gieson's stain and fibroblast growth factor (FGF) and examined under a light microscope. The numbers of lymphocytes, collagen fibers and FGF-positive cells were determined in all three groups. RESULTS: There was no significant difference in the numbers of collagen fibers between the groups (p > 0.05). No significant differences were found in the numbers of lymphocytes and FGF-positive cells between the titanium and vitallium groups (p > 0.05). The differences in the numbers of lymphocytes and FGF-positive cells between the control and other groups were found to be significant (p <0.05). PMID- 15799573 TI - Expression of Clara cell secretory protein in experimental otitis media in the rat. AB - CONCLUSION: These results suggest that CCSP is upregulated in OME and may play a protective role in the pathogenesis of OME. OBJECTIVE: Clara cell secretory protein (CCSP) is an abundant 16-kDa homodimeric protein and is secreted by non ciliated secretory epithelial cells in the lung. It has an important protective role against the intrapulmonary inflammatory process. The aim of this study was to investigate the expression of CCSP in endotoxin-induced otitis media with effusion (OME) in the rat. MATERIAL AND METHODS: We instilled endotoxin and saline (control) into the middle ear cavity of the rat. Middle ear mucosa were taken at 0, 1, 3, 6 and 12 h and 1, 3, 7 and 14 days, and the expression of both CCSP mRNA and protein were then evaluated using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry. RESULTS: RT-PCR revealed that the expression of CCSP was first identified at 1 h after endotoxin instillation, was dramatically increased between 1 h and Day 1, with maximal expression at 12 h, and then decreased after Day 3. The expression pattern of CCSP protein identified by means of Western blotting was similar to the CCSP mRNA patterns observed using RT-PCR. Expression of CCSP at both mRNA and protein levels was not detected in either normal middle ear mucosa or saline-instilled middle ear mucosa. Immunohistochemistry revealed that some epithelial cells in the middle ear mucosa were stained. PMID- 15799574 TI - Symptomatic high frequency/acceleration vestibular loss: consideration of a new clinical syndrome of vestibular dysfunction. AB - CONCLUSION: Symptomatic high frequency/acceleration vestibular loss is a distinct clinical entity that can be missed on conventional ENG with caloric testing. Under certain circumstances, symptomatic patients with a high frequency/acceleration vestibular loss should undergo an MSSC study for confirmation, if required. OBJECTIVE: To document that normal electronystagmography (ENG) with conventional bithermal caloric testing is inadequate for diagnosing clinically significant high frequency/acceleration vestibular loss. MATERIAL AND METHODS: Patients with clinical symptoms and signs of persistent peripheral vestibular dysfunction despite normal conventional bithermal caloric testing on ENG underwent high frequency/acceleration horizontal magnetic scleral search coil (MSSC) eye movement studies. The clinical findings and results from audiometric tests, conventional ENG with bithermal caloric tests and MSSC tests were reviewed. RESULTS: Eleven patients were identified as having an abnormal MSSC study, indicating a high frequency/acceleration vestibular loss consistent with their clinical history despite normal or equivocal bithermal caloric responses on conventional ENG. Although valuable, ENG caloric testing evaluates lateral semicircular canal function and should be considered a non physiological test primarily of low frequency vestibular function. High frequency/acceleration head thrust testing clinically detected a "high frequency/acceleration vestibular loss" in 8/11(72.7%) cases. PMID- 15799575 TI - Quality of life in elderly patients with dizziness: analysis of the Short-Form Health Survey in 197 patients. AB - CONCLUSION: Elderly patients with chronic dizziness had a poor HRQoL, which could be successfully assessed using the SF-36, a generic health-status instrument. The frequency of dizziness and comorbid psychological distress were predictive of impairment in QoL. OBJECTIVES: To evaluate health-related quality of life (HRQoL) and its relationship to characteristics of dizziness in elderly persons with chronic dizziness. MATERIAL AND METHODS: A standardized dizziness questionnaire was used to evaluate characteristics of dizziness as well as medical, functional and demographic data in 197 consecutive patients with chronic dizziness aged > or =60 years. HRQoL was evaluated using the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) and the Hospital Anxiety and Depression Scale (HADS). RESULTS: Compared with normative data, results from patients with dizziness indicated a pervasive multidimensional decline in SF-36 scores. This decline was most notable in terms of role limitations in the physical and emotional dimensions. The frequency of attacks of dizziness correlated with perceived disability. Patients with chronic dizziness also had great psychological distress, as measured by the HADS score, which had a detrimental influence on their QoL. PMID- 15799576 TI - Influence of visually induced self-motion on postural stability. AB - CONCLUSION: Our results indicate that the illusion of self-motion is a significant factor leading to spatial disorientation. OBJECTIVE: Under normal circumstances, self-motion is perceived in response to motion of the head and body. However, under certain conditions, such as virtual reality environments, visually induced self-motion can be perceived even though the subject is not actually moving, a phenomenon known as "vection". The aim of this study was to examine the possible influence of illusory self-rotation (circular vection) on postural adjustments. MATERIAL AND METHODS: The subjects were 10 young females with no history of ocular or vestibular disease. Video-motion analysis was applied to measure postural movements during vertical optokinetic stimulation. RESULTS: For most subjects, movement of the visual surroundings induced head and body displacements in the same direction as that of the visual stimulus, regardless of the onset of self-motion perception. However, there was a significant increase in postural instability after the subjects began to perceive false self-motion in the opposite direction to that of the visual stimulus. PMID- 15799577 TI - Immunohistochemical localization of 3-nitrotyrosine in the nasal respiratory mucosa of patients with vasomotor rhinitis. AB - CONCLUSION: This study demonstrates that, in the nasal respiratory mucosa of patients with vasomotor rhinitis, oxidative stress following peroxynitrite formation is confined to the respiratory epithelium. This suggests that the role of peroxynitrite in vasomotor rhinitis differs from its role in other diseases of the respiratory tract. The results of this study also support the concept that different pathogenetic mechanisms are probably involved in vasomotor rhinitis. OBJECTIVE: Previous studies indicated that nitric oxide (NO) is involved in the pathogenesis of vasomotor rhinitis, strong expression of NO synthase being detected in the smooth muscle cells of the cavernous sinuses and in the respiratory epithelium. However, most adverse effects of high levels of NO originate from the reaction of NO with superoxide anions to form peroxynitrite. Therefore, in this study we evaluated the involvement of peroxynitrite in the pathogenesis of vasomotor rhinitis. MATERIAL AND METHODS: Sites of peroxynitrite formation were identified by immunolabelling for 3-nitrotyrosine (3NT), its footprint in tissues. Samples of nasal mucosa were obtained from vasomotor rhinitis patients and from control subjects who had undergone corrective surgery of the nasal septum. All samples were obtained by reduction of the inferior turbinate. RESULTS: Examination of specimens from vasomotor rhinitis patients revealed that 3NT is absent in epithelium with a normal appearance, cells of the subepithelial connective tissue, the glands and the blood vessels, including the cavernous sinuses. In contrast, intense 3NT immunolabelling was found in the disrupted respiratory epithelium. 3NT was not present in any of the specimens from control subjects. PMID- 15799578 TI - Comparison between polymerase chain reaction and fungal culture for the detection of fungi in patients with chronic sinusitis and normal controls. AB - CONCLUSION: PCR using panfungal gene primers is a more sensitive method for fungus detection than fungus culture, both in patients with chronic sinusitis and in normal controls. The presence of fungi alone, however, was insufficient to implicate them as pathogens in chronic sinusitis. OBJECTIVE: Previous findings have suggested that polymerase chain reaction (PCR)-based methods are more sensitive and reliable than conventional culture methods for the detection of fungal DNA. We therefore compared these methods in 82 patients with chronic sinusitis and 40 normal controls. MATERIAL AND METHODS: The noses of the subjects were irrigated with sterile saline, and the samples collected. The sediment from each irrigation was used for fungus culture and PCR analysis. RESULTS: PCR analysis using panfungal gene primers showed that 76/82 (92.5%) patients with chronic sinusitis and 39/40 (97.5%) normal controls were positive. In contrast, fungus cultures were positive in 19/82 (23.2%) patients with chronic sinusitis and 12/40 (30.0%) normal controls. We observed no significant between-group differences in the prevalence of fungus or in the fungal species detected. PMID- 15799580 TI - Hoarseness and misdirected swallowing before and after antireflux surgery. AB - CONCLUSION: Patients with hiatus hernia can be relieved from H, MSL and MSN by hiatus hernia repair. OBJECTIVE: It has been hypothesized that respiratory symptoms in patients with gastro-oesophageal reflux disease (GORD) may, in some cases, be due to misdirected swallowing as a consequence of defective opening of the upper oesophageal sphincter. The aim of this study was to investigate whether patients with hiatus hernia are relieved from symptoms of misdirected swallowing to the larynx (MSL) and nose (MSN), as well as hoarseness (H), as a result of hiatus hernia repair. MATERIAL AND METHODS: A questionnaire concerning symptoms of H, MSL and MSN was administered to 90 patients under investigation for hiatus hernia repair before and after surgery. RESULTS: Before surgery, MSL occurred in 30% of patients, MSN in 30% and H in 25%. These symptoms were significantly interrelated (p <0.008). After antireflux surgery, all symptoms were significantly reduced (p <0.001). Symptom reduction was not related to the weight of the patients. PMID- 15799579 TI - MUC8 as a ciliated cell marker in human nasal epithelium. AB - CONCLUSIONS: This study indicates that MUC8 protein is expressed in ciliated cells from human nasal epithelial cells and is upregulated by IL-1beta treatment. These results suggest that MUC8 gene and protein expression levels could be used as a ciliated cell marker in human nasal epithelium. OBJECTIVES: To examine MUC8 mRNA expression patterns according to the mucociliary differentiation of normal human nasal epithelial (NHNE) cells, and to investigate the localization of MUC8 proteins in nasal polyps. MATERIAL AND METHODS: Passage-2 NHNE cells were cultured using an air-liquid interface technique. On Days 2, 7, 14 and 28 after confluence, ciliated cells were counted by means of cytospin slide immunostaining using H6C5 and beta-tubulin, and MUC8 mRNA levels were determined using real-time quantitative polymerase chain reaction (PCR). After synthesizing polyclonal anti MUC8 peptide antibodies, MUC8 immunostaining was performed using nasal polyps. MUC8 mRNA and protein levels were determined in NHNE cells treated with IL-1beta (10 ng/ml for 24 h) using reverse transcriptase-PCR and Western blot analysis. RESULTS: The increases in the number of ciliated cells and the MUC8 gene expression level with increasing culture time in the NHNE cells were quite similar. MUC8 was expressed in the ciliated cells of human nasal polyps. The MUC8 protein level and the mRNA level were upregulated as a result of IL-1beta treatment. PMID- 15799581 TI - Cholesterol granuloma of the temporal bone: a pathologic designation or a clinical diagnosis? PMID- 15799582 TI - Supracricoid laryngectomy with cricohyoidopexy for recurrence of early-stage glottic carcinoma after irradiation. Long-term oncological and functional results. AB - CONCLUSIONS: These results suggest that, in selected cases, SCL-CHP may be used to treat laryngeal carcinomas after radiation failure, with good oncological and functional results. OBJECTIVES: Radiotherapy and surgery are believed to be equally effective and highly successful in the management of T1-staged glottic carcinomas. An almost normal post-therapy voice is considered the main advantage of irradiation over the surgical approach. On the other hand, when a tumour recurs after radiotherapy, it is more likely to extend beyond its original site, making total laryngectomy necessary in most cases. However, in selected cases, conservative laryngeal surgery is possible. At present only a few reports in the literature have documented the oncological and functional outcome of supracricoid laryngectomy with cricohyoidopexy (SCL-CHP) in the treatment of laryngeal recurrence after irradiation. MATERIAL AND METHODS: We describe seven cases of rT2-T3 laryngeal squamous cell carcinomas that recurred after radiotherapy and were treated with salvage SCL-CHP between 1989 and 1997 at the Department of Otolaryngology, University of Ferrara. The disease-free interval following initial radiotherapy ranged from 13 to 132 months (mean 54 months) in 6 patients; 1 patient underwent 2 courses of irradiation treatment 103 and 8 months prior to surgery. RESULTS: The mean post-SCL-CHP follow-up period exceeded 10 years (range 72-173 months). Only the patient who underwent 2 courses of radiation therapy before surgery experienced laryngeal recurrence 48 months after surgery. All patients were decannulated and recovered the ability to swallow. Vocal quality was significantly rough and breathy but was satisfactorily intelligible in all patients. Four patients had wound healing problems; 1 of them developed a laryngotracheocutaneous fistula 15 months after SCL-CHP as a consequence of chondronecrosis, prompting total laryngectomy. PMID- 15799583 TI - CYP17 and tumor necrosis factor-alpha gene polymorphisms are associated with risk of oral cancer in Chinese patients in Taiwan. AB - CONCLUSION: Patients who carry the T/T homozygote of CYP17 C/T gene polymorphism may have a higher risk of developing oral cancer. OBJECTIVE: Cancer of the oral cavity is the most commonly seen malignancy in Taiwan, and its rising incidence poses a formidable challenge to oncologists. The CYP17 gene encodes P450c17alpha, an enzyme involved in the metabolism of steroid hormones. Tumor necrosis factor alpha (TNF-alpha) is one of the cytokines produced by macrophages, and its function has been postulated to play a role in cancer formation. We investigated whether polymorphisms of CYP17 and TNF-alpha genes are associated with oral cancer. MATERIAL AND METHODS: Polymorphisms of CYP17 and TNF-alpha genes were detected by polymerase chain reaction-based restriction analysis in 137 patients with oral cancer and 102 normal controls. RESULTS: The results for the CYP17 gene revealed a significant difference between oral cancer patients and normal controls (p =0.0063), but there was no significant difference for the TNF-alpha gene (p =0.4753). PMID- 15799584 TI - Intubation laryngeal mask: atraumatic diagnostic tool in suspension laryngoscopy. AB - CONCLUSION: The new technique of ILM-guided vocal cord biopsy and APC is safe, cost-effective and non-invasive. It provides excellent airway control, adequate exposure of the vocal cords and effective treatment of laryngeal bleeding. OBJECTIVE: Suspension microlaryngoscopy is a standard diagnostic procedure for vocal cord biopsy. In experienced hands failure of the procedure is extremely rare and may indicate a need for more invasive techniques. The aim of this study was to present a new technique of laryngoscopy/vocal cord biopsy and to review the relevant literature. MATERIAL AND METHODS: After induction of general anesthesia and preoxygenation an intubation laryngeal mask (ILM) was inserted. A flexible bronchoscope was passed through the ILM and an anterior lesion was identified at the vocal cord. Biopsies were taken with forceps inserted through the instrument's working channel. The procedure was performed using video endoscopic guidance. Bleeding from the wound surface was adequately treated with argon-plasma coagulation (APC). RESULTS: Identification of the vocal cords was readily accomplished using the ILM. Only four literature reports matched our search criteria; all used the standard laryngeal mask or other instruments such as a laser or did not use the procedure for definitive therapy. PMID- 15799585 TI - Thymic cyst in the neck. AB - A rare case of a thymic cyst in the neck containing both thymus and parathyroid tissue in a 7-year-old boy is presented. The clinical presentation, diagnostic evaluation, surgical management and histopathological features are described. The embryology of cervical thymic cysts and the differential diagnosis of cystic neck masses in children are briefly reviewed. The diagnosis is seldom made preoperatively. Surgical resection is the treatment of choice for definitive diagnosis, resolution of symptoms and cure. PMID- 15799586 TI - An assessment of risk associated with the photography of microbiological materials. AB - This study investigated risks associated with the photography of materials that are a potential source of infection, the extent to which medical illustrators are aware of these risks, the relevant government regulations and guidance, and whether recommended control measures are adopted. To seek information about current practice, a questionnaire was sent to randomly selected Medical Illustration Units across the United Kingdom. The data indicated that a significant proportion of medical illustrators routinely photograph materials that could be a source of infection, but that a high percentage know little about the specific regulations and guidance which govern the handling of such materials. Apparent deficiencies in safety procedures currently applied in photographing microbiological materials were identified; recommendations that address these problems at little or no cost are proposed. PMID- 15799587 TI - Setting up a store-and-forward telemedicine service for acute trauma in a hospital trust. AB - Telemedicine is a valuable, but as yet underused, resource for the delivery of health-care to patients. This paper describes the development of a new store-and forward telemedicine service at the Queen Victoria Hospital, which is a tertiary referral centre for plastic surgery in the United Kingdom. The practical requirements of such a system, and the technical and legal difficulties experienced during the initial stages, are also discussed. PMID- 15799588 TI - An Intranet-based clinical imaging service. AB - This paper explains how the Photographic Department at The Queen Victoria Hospital, East Grinstead, has transformed its clinical photography service through the use of digital technology. The Department now plays a more proactive role in patient care, and its intranet-based digital imaging system has been recognized as an area of best practice by The Department of Health Information Policy Unit and by the Medicines and Healthcare Products Regulatory Agency. PMID- 15799589 TI - Contact dermatitis. PMID- 15799590 TI - Accessible Web design. AB - As part of the Institute of Medical Illustrators' (IMI) scheme for continuing professional development (CPD), worksheets will be published at regular intervals in this Journal. These are designed to provide the members of IMI with a structured CPD activity that offers one way to earn credits. It is recognized that this worksheet requires some time spent undertaking the exercises. This activity has been tested, and hours have been allocated to individual tasks (see clocks), but these are intended only as a guide. The answers to the questions, along with any notes and reflections you make or other publications you find, should be kept in your CPD portfolio. PMID- 15799591 TI - Tobacco risk perceptions and behavior: implications for tobacco control. PMID- 15799592 TI - Do smokers want to know more about the cigarettes they smoke? Results from the EDUCATE study. AB - The present study (a) assessed smokers' receptivity to receiving information about the product features of their cigarette brand, (b) tested whether the use of targeted (personalized), brand-specific information affected participants' attention to the information, and (c) tested whether attention to the targeted information affected participants' beliefs about the product features and their smoking behavior. The study population included current cigarette smokers who called the New York State Smokers' Quit Line seeking assistance to stop smoking in February and March 2003. Subjects were randomized to one of three experimental groups. Group 1 received telephone counseling and the quit line's stop-smoking booklet, which included information on ingredients found in cigarettes. Group 2 received the same intervention as Group 1 plus a basic brochure with a generic cover. Group 3 received the same intervention as Group 2 except that the cover to the brochure was targeted to individual cigarette brand and type. All smokers who called the quit line were receptive to receiving information about their cigarette brand. In a 6-week follow-up interview, 60% of those who received the targeted product information brochure recalled receiving it vs. 51% of those who received the identical guide with the nontargeted cover. Recall of the material discussed in the brochure was slightly higher (not statistically significant) among subjects who received the brochure with the targeted cover compared with the same brochure with a basic cover. Regardless of whether the brochure was targeted, smokers' beliefs about different product features or their smoking behavior were not affected measurably, although those who reported reading some or all of the brochure had higher levels of awareness regarding low-tar, filtered, and no-additive cigarettes. Smokers are receptive to receiving information about their cigarette brand, but either persistent efforts or possibly more potent interventions to personalize the information are needed to ensure that they recall information about the cigarette brand they smoke. PMID- 15799593 TI - Stop-smoking medications: who uses them, who misuses them, and who is misinformed about them? AB - This study assessed smokers' beliefs about nicotine and the safety of nicotine medications and examined how these beliefs influence the use of nicotine medications. The data for this paper came from a nationally representative, random-digit-dialed telephone survey of 1,046 adults (18 years of age and older) current cigarette smokers conducted between May and September 2001. Respondents were questioned about their use of stop smoking medications, beliefs about nicotine, and the safety/efficacy of nicotine medications. Nearly all adult smokers in our survey had heard of nicotine patches (97%) or gum (94%), with lower levels of awareness reported for the nicotine inhaler (41%), and nasal spray (9%). Thirty-eight percent of smokers had previously used nicotine medications, with the nicotine patch being the most commonly used medication. The data reveal that most smokers are misinformed about the health risks of nicotine and the safety/efficacy of nicotine medications. Approximately half incorrectly reported that the reduction in nicotine in cigarettes has made cigarettes less dangerous to health and only one-third correctly reported that nicotine patches were less likely to cause a heart attack than smoking cigarettes. Smokers who were more knowledgeable about the health risks of nicotine and the safety and efficacy of nicotine medications were more likely to report past use of nicotine medications. Misperceptions about the health risks of nicotine and the safety/efficacy of nicotine medications may discourage some smokers from considering the use of these medications to help them stop smoking. PMID- 15799594 TI - Use of and beliefs about light cigarettes in four countries: findings from the International Tobacco Control Policy Evaluation Survey. AB - This study examined reported use of, and beliefs about, so-called light cigarettes among adult smokers in four countries: Australia (Aus), Canada (Can), the United Kingdom (U.K.) and the United States (U.S.). The method used was parallel telephone surveys among 9,046 smokers across the four countries. The results indicated that more than half of all smokers in each country except the U.K. reported smoking light cigarette brands. A majority of smokers surveyed in each country except Canada continue to believe that light cigarettes offer some health benefit compared to regular cigarettes (Canada 43%, U.S. 51%, Australia 55%, U.K. 70%). A majority of smokers in all four countries believed that light cigarettes are smoother on the throat and chest than regular cigarettes. Predictors of use of light cigarettes and beliefs about possible benefits were very similar in the four countries. These results demonstrate an ongoing need for public education about why light cigarettes do not reduce harm and do not make quitting easier. The results provide further evidence for the need for regulatory measures in all four countries to prohibit the use of misleading light and mild descriptors including package imagery in product marketing (as prescribed in Article 11 of the Framework Convention on Tobacco Control), abandon the use of standard FTC/ISO tar and nicotine yields as consumer information, and adopt policies to regulate deceptive design features of cigarettes, such as ventilated filters. PMID- 15799595 TI - What do Marlboro Lights smokers know about low-tar cigarettes? AB - The present study examined the extent to which Marlboro Lights smokers perceive lower health risks associated with using a low-tar cigarette and the extent to which they are aware of filter vents in their cigarettes. The data for this study came from a nationally representative random-digit-dialed telephone survey of 1,046 adult current cigarette smokers (aged 18 years or older) conducted between May and September 2001. Respondents were asked about the brand and type of cigarettes smoked, beliefs about the health benefits associated with low-tar and filtered cigarettes, and awareness of ventilation holes in the filters of their cigarettes. Marlboro Lights was the most popular brand, smoked by 19% of survey respondents. Only 32% of Marlboro Lights smokers reported that the filters on their cigarettes were ventilated. Many Marlboro Lights smokers believed incorrectly that lower tar, light, and ultra-light cigarettes were less harmful compared with higher tar, full-flavored cigarettes. For example, only 11% of Marlboro Lights smokers knew that the tar delivery of a light cigarette was about the same as that of a full-flavored cigarette. The responses of Marlboro Lights smokers to questions about the hazards of low-tar cigarettes were similar to those expressed by smokers of other cigarette brands. The data presented in this paper reveal that smokers of the leading light cigarette brand sold in the United States today, Marlboro Lights, are for the most part unaware of filter vents in the cigarettes they smoke and are misinformed about the health risk of using low tar and filtered cigarettes. PMID- 15799596 TI - Are smokers adequately informed about the health risks of smoking and medicinal nicotine? AB - The present study assessed smokers' beliefs about the health risks of smoking and the benefits of smoking filtered and low-tar cigarettes, and their awareness of and interest in trying so-called reduced-risk tobacco products. Results were based on a nationally representative random-digit-dialed telephone survey of 1,046 adult (aged 18 years or older) current cigarette smokers. Data were gathered on demographic characteristics, tobacco use behaviors, awareness and use of nicotine medications, beliefs about the health risks of smoking, content of smoke and design features of cigarettes, and the safety and efficacy of nicotine medications. In addition, respondents were asked about their interest in and perceived ability to stop smoking and about their desire for more information about the health risks of smoking. Smokers were least knowledgeable about low-tar and filter cigarettes (65% of responses were incorrect or "don't know") and most knowledgeable about the health risks of smoking (39% of responses were incorrect or "don't know"). The smokers' characteristics most commonly associated with misinformation when all six indices were combined into a summary index were as follows: those aged 45 years or older, smokers of ultralight cigarettes, smokers who believe they will stop smoking before they experience a serious health problem caused by smoking, smokers who have never used a stop-smoking medication, and smokers with a lower education level. Those who believed they would stop smoking in the next year were more knowledgeable about smoking. Some 77% of respondents reported a desire for additional information from tobacco companies on the health dangers of smoking. The present findings demonstrate that smokers are misinformed about many aspects of the cigarettes they smoke and stop-smoking medications and that they want more information about ways to reduce their health risks. PMID- 15799597 TI - The near-universal experience of regret among smokers in four countries: findings from the International Tobacco Control Policy Evaluation Survey. AB - Regret may be a key variable in understanding the experience of smokers, the vast majority of whom continue to smoke while desiring to quit. We present data from the baseline wave (October-December 2002) of the International Tobacco Control Policy Evaluation Survey, a random-digit-dialed telephone survey of a cohort of over 8,000 adult smokers across four countries--Canada, the United States, the United Kingdom, and Australia--to estimate the prevalence of regret and to identify its predictors. The proportion of smokers who agreed or agreed strongly with the statement "If you had to do it over again, you would not have started smoking" was extremely high--about 90%--and nearly identical across the four countries. Regret was more likely to be experienced by older smokers, women, those who had tried to quit more often, those who perceived quitting as conferring benefits, those with higher levels of perceived addiction, those who worried about future damage to health, those who perceived smoking as lowering their quality of life, those who perceived higher monetary costs of smoking, and those who believed that smoking is not socially acceptable. This predictive model was the same in all four countries. Regret is thus a near-universal experience among smokers in all four countries, and the factors that predict regret are universal across these four countries. Among other implications for cessation treatment and smoking prevention, this near universality of regret casts doubt on the view of some policy analysts and economists that the decisions to take up and continue smoking are welfare-maximizing for the consumer. PMID- 15799598 TI - Smokers' responses to advertisements for regular and light cigarettes and potential reduced-exposure tobacco products. AB - This study examines smokers' responses to advertisements for potentially reduced exposure tobacco products (PREP), light cigarettes, and regular cigarettes. A convenience sample of 600 adult smokers reviewed one actual advertisement for each type of product. Smokers ranked the products on health risk, amount of tar, and carcinogenicity, and identified the messages they perceived the advertisements to convey. Smokers perceived PREP products as having lower health risks (mean = 5.4 on a scale of 1-10) and carcinogens (6.6) than light cigarettes (5.8 and 6.9, respectively, p < .001), and lights as having lower health risks and carcinogen levels than regular cigarettes (8.2 and 8.8, respectively, p <.001). The average PREP rating for level of tar (5.3) was not significantly less than the light mean of 5.4, but both were significantly less than the regular mean of 8.4 (p <.001). Although no advertisements explicitly said that the products were healthy or safe, advertisements for PREP products and light cigarettes were interpreted as conveying positive messages about health and safety. Most smokers believed that claims made in cigarette advertisements must be approved by a government agency. The results indicate that advertisements can and do leave consumers with perceptions of the health and safety of tobacco products that are contrary to the scientific evidence. Explicit and implicit advertising messages may be strengthened by the perceived government endorsement. This supports the Institute of Medicine's recommendation to regulate the promotion, advertising, and labeling of PREP tobacco products and light cigarettes. Effective regulation may need to focus on consumer perceptions resulting from advertisements rather than the explicit content of advertising text. PMID- 15799600 TI - Perceptions about cigarette smoking and risks among college students. AB - The objective of the present study was to describe how college students perceive the risks of cigarette smoking and addiction to nicotine. Data came from a self administered survey of 1,020 college students enrolled in two 4-year liberal arts colleges in the United States. The survey was conducted in the fall of 2001. Smokers and nonsmokers differed markedly in their perceptions about the health risks associated with short-term exposure to smoking. College students in this sample who smoked did not fully comprehend the risks associated with smoking. Smokers were half as likely as nonsmokers to believe that there are health risks from smoking only on weekends or a couple of days a week. Anti-tobacco messages for young adult smokers need to communicate more effectively the concept that each cigarette they smoke is doing them damage. PMID- 15799599 TI - Predictors of cessation in a cohort of current and former smokers followed over 13 years. AB - The present study attempted to identify predictors of smoking cessation in a cohort of cigarette smokers followed over 13 years. Data are reported on 6,603 persons who resided in one of 20 U.S. communities involved in the National Cancer Institute's Community Intervention Trial for Smoking Cessation (COMMIT) study, were current smokers in the COMMIT trial in 1988, and completed detailed tobacco use telephone surveys in 1988, 1993, and 2001. A person was classified as a former smoker if at the time of follow-up he or she reported not smoking for at least 6 months prior to the interview. Reasons and methods for quitting also were assessed in 1993 and 2001. Among smokers in 1988, 24% had stopped smoking by 1993 and 42% were not smoking by 2001. The most frequently cited reasons for quitting were health and cost reasons, while assisted methods to quit were more common in more recent years. Measures of nicotine dependence were much more strongly associated with cessation than measures of motivation. Other predictors included male gender, older age, higher income, and less frequent alcohol consumption, although the gender effect no longer existed when cessation from cigarettes as well as other tobacco products was considered as the outcome. The present study shows that nicotine dependence is a major factor predicting long-term cessation in smokers. This finding has implications for tobacco control policy and treatment approaches. PMID- 15799601 TI - Accuracy and optimism in smokers' beliefs about quitting. AB - Data collected in two nationwide surveys (N=361 and N=788) provide insight into the extent to which smokers recognize the ease of addiction and the difficulty of quitting. Smokers generally acknowledge that addiction occurs quickly and that smokers have considerable difficulty quitting. Youth smokers, however, generally believe that they would have less difficulty than other smokers and that they are less addicted than the average smoker. Adult smokers tend to say they are not different from other smokers in addiction or ability to quit. Nevertheless, both youth and adult smokers who want to quit greatly overestimate the likelihood that they will succeed in the coming year. Few smokers think about how long they will smoke when they first begin, and the reluctance to believe that they could have serious difficulty quitting may help in the progression from experimental to established smoker. PMID- 15799602 TI - Evolution and revolution at the menopause. PMID- 15799603 TI - Menopause: why does fertility end before life? AB - Menopause is associated with an ultimate cessation of child-bearing potential. Medical research on menopause focuses mostly on the underlying physiological changes associated with menopause. By contrast, evolutionary biologists are interested in understanding why women lose their potential to reproduce before the end of their lives. Evolution by natural selection predicts that the behaviors that we observe today are products of generations of selection on the genes that govern those behaviors. Since one would expect an individual reproducing throughout its life to produce more offspring than an individual stopping early, one would seldom expect genes for menopause to be selected for during our evolutionary past. This article discusses how menopause and prolonged lifespan might be explained by evolutionary theory, and highlights some angles for future research. PMID- 15799604 TI - Guidelines for hormone treatment of women in the menopausal transition and beyond. PMID- 15799605 TI - Hormones and depression in women. AB - The biological plausibility for the effect of sex hormones on the central nervous system is now supported by a considerable amount of clinical data. This critical review guides the reader through the plethora of data, from the earliest reports of menstrual madness in the nineteenth century to neurobiological work in the new millennium. It illustrates through the scientific evidence base that, although the effect of estrogen on the central nervous system, particularly on mood and depression, remains a controversial area, there is now considerable evidence for the psychotherapeutic benefits of estrogens in the triad of hormone-responsive depressive disorders: postnatal depression, premenstrual depression and perimenopausal depression. The article also reviews the compelling data that testosterone supplementation has positive effects for depression, libido and energy, particularly where patients have only partially responded to estrogen therapy. PMID- 15799606 TI - Transdermal hormone therapy: gels and patches. AB - Hormone therapy (HT) in the climacteric has a number of beneficial effects including mitigation of climacteric symptoms and prevention of osteoporosis. Administration of HT via the transdermal route avoids hepatic first-pass metabolism and therefore the high plasma levels of estrogen metabolites that are associated with oral administration. Patch formulations have traditionally been the most common form of transdermal HT. However, as patches may be associated with local skin reactions, gel formulations have been developed in an attempt to improve acceptability and compliance with transdermal HT. Patch and gel formulations are equally as effective in treating climacteric symptoms and improving bone mineral density, and the effects are comparable to those achieved by oral HT. PMID- 15799607 TI - A longitudinal population study of climacteric symptoms and their treatment in a random sample of Swedish women. AB - OBJECTIVES: To assess, first, the prevalence and severity of symptoms associated with the climacteric period and their treatment and, second, the prevalence of exercise, smoking and body weight in a population-based sample of Swedish women. MATERIAL AND METHODS: Prospective, longitudinal cohort study, initiated in 1992, in women aged 46, 50, 54, 58 and 62 years with a follow-up 6 years later. Information was obtained from the same women (n=3816) on both occasions using a postal questionnaire regarding sociodemographic variables, general and reproductive health, the occurrence of climacteric symptoms and their severity, and the use of hormone replacement therapy (HRT). RESULTS: The prevalences of climacteric symptoms were as follows (1992/1998): vasomotor symptoms, 52%/62%; depression/irritability, 57%/65%; sleeping disturbances, 51%/69%; muscle/joint pain, 55%/70%; and loss of libido, 38%/57%. HRT with medium-potency estrogens was currently being used by 34% (1992: 14%), and 12% (1992: 8%) were using low potency estrogens. The maximum prevalence of HRT (medium-potency estrogens) use was found in the 56-year-old group, at 46% (1992: 25% in the 54-year-old group). Body mass for the whole group had increased from 66.3 to 68.9 kg. Exercise was more frequent in all age groups in 1998 compared to 1992. There was a decrease in current smokers from 32 to 26% between the two periods. Compared with 1992, the women in all five birth cohorts considered themselves to be less healthy and quality of life had decreased for the whole group. CONCLUSIONS: The prevalence of symptoms associated with the climacteric period and the use of HRT had increased markedly in this longitudinal study of the same women followed between 1992 and 1998. During the same period, smoking decreased, while body weight and exercise frequency increased. PMID- 15799608 TI - Effect of hormone replacement therapy on lipids and left ventricular function in postmenopausal smokers. AB - OBJECTIVE: Menopause and smoking have negative effects on the cardiovascular system. The study was planned to investigate the influence of oral hormone replacement therapy (HRT) on heart function and lipids in postmenopausal smokers. METHODS: Lipid levels and left ventricular systolic and diastolic function by means of echocardiography were assessed before entering the study and at 6-month intervals during the 12 months of oral HRT in 62 postmenopausal women, 30 of whom were smokers and 32 were non-smokers. RESULTS: Oral HRT caused a significant decrease in levels of total cholesterol and low density lipoprotein (LDL) cholesterol and a significant increase in high density lipoprotein (HDL) cholesterol in non-smokers. This effect was not evident in smokers. Echocardiography revealed a significant improvement of systolic function (ejection fraction, left ventricular outflow tract velocity, forward velocity integral, acceleration time and mean systolic acceleration) and diastolic function (diastolic time, duration of the early filling phase, peak velocity of early mitral flow, and the ratio of late to early peak mitral flow) in non smokers. In smokers, a significant increase in some parameters of systolic function (ejection fraction, acceleration time and mean systolic acceleration) and an insignificant change in diastolic function were observed. Oral HRT of 12 months' duration has very limited beneficial effects on CONCLUSION: lipids and left ventricular heart function in postmenopausal women who smoke. PMID- 15799609 TI - The menopausal transition: a 9-year prospective population-based study. The Melbourne Women's Midlife Health Project. AB - OBJECTIVES: To describe the natural history of the menopause in Australian-born women. To determine the hormonal changes relating to the menopausal transition (MT) and how these affect quality of life, bone mineral density, body composition, cardiovascular disease (CVD) risk and memory. DESIGN: A 9-year prospective, observational study of a population-based sample of 438 Australian born women aged 45-55 years at baseline. By the 9th year, the retention rate was 88%. Interviews, blood sampling, menstrual calendars, quality of life and physical measures were taken annually, and bone mineral density was measured bi annually. RESULTS: The late MT coincides with changes in estradiol, follicle stimulating hormone, and free testosterone index, decreases in bone density and mastalgia, and increases in central adiposity, vasomotor symptoms, insomnia and vaginal dryness. Levels of total testosterone and dehydroepiandrosterone sulfate are unchanged by the MT. An increase in CVD risk was associated with increases in weight and free testosterone index and a decrease in estradiol. Depressed mood is increased by symptoms and by stressors occurring in the MT. Sexual functioning significantly deteriorates with the MT and aging, but relational factors have major effects. Menstrual cycles became more variable and longer closer to the final menstrual period. CONCLUSIONS: As hormonal changes during the MT directly or indirectly adversely affect quality of life, body composition and CVD risk, maintenance of health parameters in the premenopausal years is crucial for a healthy postmenopause. PMID- 15799610 TI - Smoking reduces breast tenderness during oral estrogen-progestogen therapy. AB - OBJECTIVE: We wished to study the influence of smoking upon the occurrence of breast tenderness during oral estrogen-progestogen therapy (EPT). METHODS: Data from 626 healthy postmenopausal women participating in three double-blind, randomized, controlled long-term trials of EPT versus placebo were included. The studies covered sequential, continuous and interrupted regimens of estradiol opposed by a selection of progestins. All studies were mono-center studies and performed in the period 1988-1997. Data on breast tenderness were collected from adverse event reporting and information on smoking status was obtained by interview. RESULTS: Smoking was associated with an earlier age at menopause (difference: 1.3 years, p < 0.001) and a slightly lower body mass index (difference: 0.8 kg/m2, p < 0.01). Smoking women in the EPT groups had significantly lower on-treatment estradiol levels compared to non-smoking women (p < 0.001), whereas no differences were observed in the placebo group. In parallel, the incidence of breast tenderness during EPT was reduced by about one half (p < 0.001) in smokers compared to non-smokers, whereas no differences were seen on placebo. CONCLUSION: Current smoking reduces the incidence of breast tenderness in women receiving oral EPT. This may be caused by the increased degradation of estradiol during smoking. PMID- 15799611 TI - Phytoestrogens possess a weak antioxidant activity on low density lipoprotein in contrast to the flavonoid quercetin in vitro in postmenopausal women. AB - BACKGROUND: Phytoestrogens are a family of plant-derived compounds with weak estrogenic and antiestrogenic properties. The antioxidant capacity of phytoestrogens has been proposed as one of the important mechanisms that explain their health benefits. OBJECTIVE: To determine the in vitro potency of three phytoestrogens, ubiquitous in food, (biochanin A, daidzein and genistein) as antioxidants of low density lipoprotein (LDL) and to compare them with the well established antioxidant actions of estradiol and quercetin, an ubiquitous flavonoid which is found in high concentration in onions, tea and berries. METHODS: LDL was isolated by ultracentrifugation from the plasma of ten healthy postmenopausal women who were not on hormone therapy. Aliquots containing 0.5 mg of protein were incubated for 4 h with CuSO4 15 micromol/l to induce oxidative stress and with one of the five compounds studied: estradiol, quercetin, biochanin A, daidzein, and genistein, in doses of 0, 5, 15, 50, 500, 1000 and 2000 micromol/l. In addition, we studied the combined effect of estradiol 1 micromol/l plus quercetin 1 micromol/l, comparing their antioxidant action with that of each compound separately. Malonaldehyde (MDE nmol/ mg protein) was measured as a marker of LDL oxidation. RESULTS: Estradiol and quercetin induced a dose-dependent decrease in MDE concentration (p < 0.01). Comparing the areas under the curve, the antioxidant effect of quercetin was 8 times higher than the one observed with estradiol (p < 0.01). A 50% decrease in MDE was reached by quercetin at a concentration of 3.4 micromol/l, estradiol at 29 micromol/l, genistein at 280 micromol/l, biochanin at 1312 mmol/l and daidzein at 8007 mmol/l. Estradiol 1 micromol/l and quercetin 1 micromol/l did not modify MDE generation separately, but, when incubated combined, there was a significant decrease of MDE (p < 0.02). CONCLUSION: The phytoestrogens studied showed a weak antioxidant activity in vitro. The flavonoid quercetin, in contrast, showed the most potent antioxidant activity in vitro, higher than estradiol. Estradiol and quercetin showed additive antioxidant activity. We speculate that different compounds with variable antioxidant effects could amplify their antioxidant capacity when acting combined. PMID- 15799613 TI - Second thoughts on the women's health initiative study: the effect of age on the safety of HRT. PMID- 15799612 TI - The acceptability of a small intrauterine progestogen-releasing system for continuous combined hormone therapy in early postmenopausal women. AB - OBJECTIVE: To assess the acceptability, ease of insertion, tolerance and associated bleeding of a novel intrauterine progestogen-releasing system, combined with transdermal estradiol, in postmenopausal women. DESIGN: An open non comparative study of 294 postmenopausal women with an intact uterus at 27 centers in six countries. METHOD: All subjects had requested treatment for menopausal symptoms and had received transdermal estradiol 50 microg/day by patch and an experimental intrauterine system (MLS) releasing levonorgestrel 10 microg/day. Details of the insertion were recorded and subsequent bleeding, side-effects and adverse events were documented on a daily diary. Endometrial thickness was measured by transvaginal ultrasound scan prior to insertion of the MLS and after 12 months. The study is of 3 years duration. This report summarizes the results after the first year. RESULTS: The median age of the subjects was 52.6 years (range 41.7-59.6 years), 90% were within 3-5 years of menopause and 78% had previously used hormone therapy (HT). The MLS was inserted at the first attempt in 297/294 (94%) subjects and was facilitated by dilatation of the cervical canal in 65 (22%) subjects. Local anesthesia was given to 30 (10%) subjects. Of the 17 with a failed first insertion, two subjects did not want a second attempt, in 14 a second attempt succeeded, facilitated by cervical dilatation in seven and local anesthetic in three subjects. There was one failed insertion. Investigators rated 80% of first insertions as easy, 14% as slightly difficult and 6% as very difficult. The second insertion was easy in 9/15 subjects, slightly difficult in 4/15 and very difficult in one (data from one subject missing). During insertion, 34% subjects had no pain, 49% said it was mild, in 15% it was moderate and in 2% the pain was severe. At 12 months, one MLS had been expelled. Spotting was the most common form of bleeding and this decreased from a median of 9 to 0 days at 4 months, although 10% of subjects continued to report spotting up to 12 months. Bleeding days were few and mainly in the first 2 months. At the end of month 11, 66.8% were amenorrheic and 87% non-bleeding. Only nine subjects discontinued because of bleeding. There was no correlation between the bleeding profiles and number of years since menopause, previous HT use or body mass index. The median endometrial thickness at entry to the study (without HT) was 2.6 mm and 3.4 mm at 12 months. Of the adverse events, after the pain associated with insertion, headache was the next most common at 13.3% and mastalgia was experienced by 7.8% of subjects. Overall, the dropout rate was low at 28 subjects (9.5%). There were favorable changes in the serum lipid profile consistent with the effects of estradiol and suggesting minimal attenuation by the intrauterine progestogen. Conclusion This interim report of a 3-year study has confirmed that the menopausal levonorgestrel intrauterine system is easy to insert and is well tolerated by postmenopausal women. PMID- 15799614 TI - Published results on breast cancer and hormone replacement therapy in the Million Women Study are correct. PMID- 15799615 TI - Effect of static magnetic fields on the amplitude of action potential in the lateral giant neuron of crayfish. AB - PURPOSE: To investigate whether exposure to static magnetic field (SMF) affects the passive properties of neurons that mediate tail-flip escape behavior in crayfish. MATERIALS AND METHODS: A permanent magnet was placed under the isolated nerve cord of crayfish to experience SMF at 4.74 to 43.45 mT intensity for various period of time (20 seconds to 3 hours). An intracellular electrode was impaled on the axon of the lateral giant neuron (LG) of the last abdominal ganglion of crayfish to record the evoked action potential (AP) and excitatory postsynaptic potential (EPSP). The amplitudes of evoked AP and EPSPs before and after SMF exposure were measured to study the effect of SMF exposure. RESULTS: The exposure to SMF increased the amplitude of AP in the LG depending upon both the intensity of field and duration of field exposure. The changes in AP by field exposure are likely to be mediated by the increasing level of intracellular Ca2+ in the LG because the chelating of intracellular Ca2+ would block the effects by SMF exposure, while the injection of Ca2+ into the LG could mimic the effects of SMF exposure. SMF exposure also increases the input resistance of the LG membrane. Therefore, the magnitude of the EPSP in LG evoked by electrical shock on the sensory nerves was found to be enhanced after SMF exposure. CONCLUSION: SMF is usually considered to be safe for the biological issues since no electrical current is induced via the Faraday effect. Our results showed that some passive membrane properties of neurons are affected by SMF exposure. The increase in magnitude of evoked AP and EPSP suggests an increase in the sensitivity of the LG neuron. These changes by SMF exposure may not necessarily to be harmful to animals; however, further study is needed to address the biological effects from SMF exposure, especially in nervous systems. PMID- 15799617 TI - Differentiation status of human squamous cell carcinoma xenografts does not appear to correlate with the repopulation capacity of clonogenic tumour cells during fractionated irradiation. AB - PURPOSE: To investigate the magnitude and kinetics of repopulation in a moderately well differentiated UT-SCC-14 human squamous cell carcinoma [hSCC] in nude mice. This question is of interest because clinical data indicate a higher repopulation capacity in those SCC that have preserved characteristics of differentiation, which appears to be in contrast to results on FaDu and GL hSCC previously reported from this laboratory. METHODS AND MATERIALS: UT-SCC-14 tumours were transplanted subcutaneously into the right hind leg of NMRI nu/nu mice. Fractionated radiation treatments were delivered, either under clamped hypoxia at 5.4 Gy/fraction or under ambient conditions (consistent with an OER of 2.7). Tumours were irradiated every day, every 2nd day, or every 3rd day with 6, 12 or 18 fractions. 1, 2 or 3 days after the last fraction, graded top-up-doses under clamped conditions were given for the purpose of estimating the 50% tumour control dose (TCD50). A total of 22 TCD50 assays were performed and analysed using maximum likelihood techniques. RESULTS: The data demonstrate a slow but significant repopulation of clonogenic cells during fractionated irradiation of UT-SCC-14 hSCC. The results under hypoxic conditions are consistent with a constant repopulation rate, with a clonogenic doubling time (Tclon) of 15.6 days (95% CI: 9.7, 21.4). This contrasts with ambient conditions where Tclon was 68.5 days (95% CI: 124, 161). Both Tclon values are longer than the 6-day volume doubling time of untreated tumours. CONCLUSIONS: Less pronounced repopulation for irradiation under ambient compared to clamped hypoxic conditions might be explained by preferential survival of hypoxic and therefore non-proliferating clonogenic cells. Taken together with previous studies on poorly differentiated FaDu and moderately well differentiated GL hSCC, the results are consistent with considerable variability in the magnitude and kinetics of repopulation in different experimental squamous cell carcinomas, and with a relationship between reoxygenation and repopulation during fractionated irradiation. The differentiation status of hSCC growing in nude mice does not to appear to correlate with the proliferative capacity of clonogenic tumour cells during treatment. The results do not support the hypothesis gained from clinical data of higher repopulation in well-differentiated tumours. PMID- 15799616 TI - Vibration as a possible explanation for putative electromagnetic field effects: a case study on marine diatoms. AB - PURPOSE: We examined the conjecture that mechanical vibration, being an uncontrolled and variable factor from one trial to another, might explain the inconsistency of results from investigations of the influence of extremely low frequency electromagnetic fields on the Ca-dependent motility of marine diatoms (Amphora coffeaeformis, strain #2038). MATERIALS AND METHODS: Owing to slight differences in culture of diatoms in previous studies, culture techniques are described in detail here. Diatoms showed maximal motility in logarithmic growth and motility was dependent on external [Ca], reducing at <.25 mM added Ca. Ninety six different vertical vibration treatments were applied to the petri dishes containing the agar on which the diatoms were placed. The envelope of amplitudes varied from 50-500 pmicro at 10Hz to 100 nm-l1mmicro at 500 Hz. RESULTS: No significant effect of the mechanical vibrations on the motility response of diatoms was observed. We were unable to impose deliberate vibrations in a horizontal direction though some component of horizontal movement was probably present in our tests. CONCLUSIONS: The results imply that the variability of earlier experiments with extremely low frequency electromagnetic fields is unlikely to be ascribable to mechanical vibration. PMID- 15799618 TI - Involvment of c-Abl in the radiation-induced inhibition of myoblast differentiation. AB - PURPOSE: While radiation-induced differentiation is common in many cell types it has been shown (Ikeda et al. 2000) that irradiation inhibits differentiation of murine myoblast cell line C2C12. To investigate the mechanisms responsible for this phenomenon we looked into the role of the non-receptor tyrosine kinase c Abl. MATERIAL AND METHODS: C-Abl activity was determined using an immune-complex kinase assay and the functional role of c-Abl activity was assessed by using the kinase inhibitor STI 571 (Gleevec) and by transfecting a dominant negative c-Abl mutant (K290R). RESULTS: We found an up-regulation of c-Abl activity 6h after differentiation induction when compared to uninduced control cells. Exposure of C2C12 cells to ionizing radiation, which inhibits myotube formation, inhibited this up-regulation. Transfected dominant negative c-Abl as well as STI 571 treatment and ionizing radiation inhibited myotube formation but did not significantly suppress the expression of the muscle-specific transcription factor Myogenin or of the differentiation marker Myosin heavy chain. CONCLUSIONS: These data suggest that c-Abl is involved in the radiation-induced inhibition of myoblast differentiation and lead to the hypothesis that c-Abl plays a role in late events during myogenic differentiation probably in the process of myoblast fusion. PMID- 15799619 TI - Analysis of alpha-particle induced chromosome aberrations in human lymphocytes, using pan-centromeric and pan-telomeric probes. AB - PURPOSE: The aim of the present study has been the evaluation of the incomplete chromosome aberrations induced after alpha-particle irradiation by the simultaneous detection of all centromeres and telomeres present in human lymphocytes. Moreover, a study on the lengths of the different acentric fragments is presented. MATERIALS AND METHODS: Attached lymphocytes were irradiated at doses of 0.2, 0.5, 0.7 and 1 Gy using a 241Am source. Flourescent in-situ hybridization (FISH) techniques were applied using pan-centromeric and pan telomeric probes. All abnormal cells were digitalised and analysed using a Cytovision FISH workstation. The description of all abnormalities observed, and the length of the acentric fragments was recorded. RESULTS: A total of 378 incomplete chromosomes plus incomplete acentrics was found. Cases with more than 92 telomeres were not detected. The ratio between total incomplete elements and multicentrics was 1.00. The total number of acentric (ace) fragments was 822; 57% of them were complete fragments ace (+,+), 26% incomplete fragments ace (+,-), and 17% interstitial fragments ace(-,-); the mean relative lengths were 2.91 +/- 0.06, 1.91 +/- 0.07 and 1.63 +/- 0.07, respectively. In all three cases a secondary peak in the length distribution was found, corresponding to a relative length between 3.5 and 4. CONCLUSION: The percentage of incomplete rejoinings is higher after alpha-particle exposure than that described previously for low linear energy transfer (LET) radiation exposures. The results seem to indicate that compared to low-LET radiation, after alpha-particle exposure centromere containing elements are more likely to be repaired.Many interstitial fragments are large linear forms that cannot be considered as non-distinguishable acentric rings. PMID- 15799620 TI - Chromosomal radiosensitivity in BRCA1 and BRCA2 mutation carriers. AB - PURPOSE: The chromosomal radiosensitivity of a selected group of familial breast cancer patients carrying a mutation in BRCA1 (n=11) or BRCA2 (n=9) and a group of healthy mutation carriers (n=12) was investigated and compared to a reference group of breast cancer patients without a BRCA1/2 mutation (n=78) and a group of healthy women carrying no mutation (n=58). MATERIALS AND METHODS: The chromosomal radiosensitivity was assessed with the G2 and the G0-micronucleus (MN)-assay on fresh blood samples and on Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines. For the MN-assay, lymphocytes were exposed in vitro to 3.5 Gy and 2 Gy 60Co gamma-rays at a high dose rate (HDR) or low dose rate (LDR). 70-h post irradiation cultures were arrested and micronuclei were scored in 1000 binucleate cells. For the G2-assay lymphocytes were irradiated in vitro with a dose of 0.4 Gy 60Co gamma-rays after 71h incubation. Cultures were arrested 90 min after irradiation and chromatid breaks were scored in 50 metaphases. RESULTS: The group of breast cancer patients with a BRCA1 or 2 mutation was on average more radiosensitive than the control group, but not different from breast cancer patients without a BRCA mutation. The radiation response of healthy BRCA1/2 carriers was not significantly different from the control group and also not different from relatives without a BRCA mutation. Comparing the radiation response in EBV cell lines derived from breast cancer patients with or without a BRCA1 mutation revealed no significant difference. CONCLUSIONS: Our results reveal that chromosomal radiosensitivity observed in breast cancer patients heterozygous for BRCA1 or 2 mutations, could not be demonstrated in healthy BRCA1/2 mutation carriers. This suggests that mutations in BRCA1 or 2 genes are not playing a main role in chromosomal radiosensitivity, this although BRCA1 and 2 are both involved in DNA repair/signalling processes. PMID- 15799621 TI - Ionizing radiation-induced chromosomal rearrangements occur in transcriptionally active regions of the genome. AB - PURPOSE: The mechanism by which ionizing radiation induces chromosomal rearrangements in mammalian cells has for long been a subject of debate. In order to dissect these events at a molecular level, we have studied the sequences involved in gamma irradiation-induced rearrangements. MATERIALS AND METHODS: An inverse polymerase chain reaction (PCR)-based methodology was used to amplify rearrangements that had occurred between one of four target regions (in or neighbouring the avian myelocytomatosis viral oncogene homologue (c-MYC), cyclin dependent kinase inhibitor 1A (CDKN1A), fibroblast growth factor receptor 2 (FGFR2), or retinoblastoma 1 (RB1) genes) and sequences elsewhere in the genome, following gamma irradiation and subsequent incubation at 37 degrees C of normal human IMR-90 fibroblasts. RESULTS: The sequences of 90 such rearrangements, including both inter- and intra-chromosomal events, have been analysed. Sequence motifs (including DNA topoisomerase recognition sites) were not found to be consistently present either at or near rearrangement breakpoint sites. Statistical analysis suggested that there was significantly more homology between the sites of DNA rearrangement breakpoints than would be expected to occur by chance, however, most DNA rearrangements showed little or no homology between the interacting sequences. The rearrangements were shown to predominantly involve transcriptionally active sequences, a finding that may have significant implications for our understanding of radiation-induced carcinogenesis. CONCLUSION: The results obtained are difficult to reconcile with most models for ionizing radiation-induced chromosomal aberration formation, but are consistent with the Transcription-Based model. PMID- 15799622 TI - Influence of heavy metals upon the retention and mobilization of polonium-210 in rats. AB - PURPOSE: To provide information about the tissue retention and mobilization of the alpha-emitting radionuclide, polonium-210 (210Po), in rats under combined exposure to heavy metal ions and the chelating agent, 2, 3-dimercaptopropane-1 sulfonate (DMPS). MATERIALS AND METHODS: Rats were pre-exposed intraperitoneally to either CdCl2 or Pb(CH3COO)2. 9 or 15 h later they received 210Po nitrate intravenously. The retention and excretion of 210Po via the urine and faeces of pre-exposed rats, as well as in pre-exposed rats treated with DMPS, were followed. The radioactivity due to 210Po in a broad spectrum of body tissues and excreta was measured by the liquid scintillation counting after sample digestion in a mixture of perchloric acid and hydrogen peroxide. The immunohistochemical localization of metallothioneins (MT) was studied using a mixture of murine monoclonal antibodies directed against MT I+II. RESULTS: The present study revealed different tissue distributions of polonium-210 in the rats pre-exposed to lead or cadmium ions when compared with that in 210Po only controls. Under combined exposure to Pb or Cd, the spontaneous excretion of 210Po was enhanced and could be further enhanced by treatment with DMPS. Treatment with this chelator was efficient even when its start was postponed until 24h after internal contamination of the body with 210Po. CONCLUSIONS: Polonium-210 is bound in vivo to binding sites on various biomolecules, among them erythrocytic enzymes and MT. This phenomenon explains the different affinity and overall distribution of 210Po in control body tissues. When the appropriate binding sites are occupied by lead or cadmium, enhanced natural excretion of polonium-210 occurs. PMID- 15799623 TI - Comparative absorption parameters of Pu and Am from PuO2 and mixed oxide aerosols measured after in vitro dissolution test and inhalation in rats. AB - PURPOSE: To compare specific absorption parameter values obtained from in vitro dissolution studies (this paper) and in vivo experiments (data published by Ramounet et al, 2000) and to determine their influence on Dose Per Unit Intake (DPUI) calculations. MATERIALS AND METHODS: Experiments were performed on plutonium oxide (PuO2) and two Mixed Oxide (MOx) preparations containing 5% Pu (w/w) made according to the industrial process in vitro using a static test and in vivo after rat inhalation. RESULTS: Behaviour of Pu and Am shows, in vitro, at shorter times, a greater rapid dissolution fraction f(r) for Pu (factor 10) and Am (factor 2) with MOx powders compared with PuO2, whereas in vivo results show a greater fraction f(r) for Pu (factor 5) and Am (factor 15) with PuO2 compared with MOx powders. This phenomenon has not been observed for slow dissolution absorption parameter s(s). The in vivo parameters for Pu and Am in these materials were very close to the default values recommended by International Commission for Radiological Protection for default Type S. CONCLUSIONS: Results obtained have shown that solubility of Pu from the mixed oxide was higher than that of Pu from PuO2. Nevertheless, no significant difference was observed between the three compounds in the corresponding dose coefficients in vivo or in vitro. Therefore, for these particular compounds, variation in the chemical composition of the aerosols had no significant influence on DPUI. Consequently, in vitro, the dissolution test can provide a good estimate of the in vivo behaviour. Studies of variation of % Pu (w/w) from MOx are in progress in our laboratory to confirm these conclusions. PMID- 15799624 TI - Seasonal changes in poisoning exposures reported to a regional poison center from coastal resort areas. AB - This study examined the role of seasonality in the reporting of poisoning exposures from geographically distinct regions, specifically from coastal resort and vacation areas. The monthly distribution of calls received by a regional poison control center from counties with popular beach and vacation resorts was compared with the monthly distribution of the overall calls to the center. A chi square goodness-of-fit test was used to determine if there was a significant difference between the monthly distribution of calls received from the resort counties and the overall calls received by the poison center. Further, exposure and information calls from the resort counties were separately examined to determine if they were equally distributed between months. The monthly distribution of calls received from coastal resort counties was significantly different from the monthly distribution of overall calls received by the center. Significantly more calls were received from the resort counties during the months of July and August at the height of the vacation season. While there was no seasonal variation in the number of information calls from these counties, the poisoning exposure calls were not equally distributed between months, as there were more such calls during the months of July and August. Seasonality appears to play a role in the number of calls received by a regional poison center from coastal areas with popular beach resorts. Poisoning exposure calls seem to increase particularly during the months of July and August. A greater effort may have to be put into activities related to poison control and prevention into such areas during the vacation season. PMID- 15799625 TI - Decreasing trend in renal disease mortality after cessation from arsenic exposure in a previous arseniasis-endemic area in southwestern Taiwan. AB - Arsenic has been well documented as a major risk factor for blackfoot disease (BFD), a unique peripheral vascular disease that was endemic in the southwestern coast of Taiwan, where residents imbibed artesian well water containing excessive amounts of arsenic for more than 50 yr. Long-term arsenic exposure has also been reported to be associated with mortality attributed to renal disease. A tap-water supply system was implemented in the early 1960s in the BFD endemic areas. Artesian well water was no longer used for drinking and cooking after the mid 1970s. The objective of this study was to examine whether mortality attributed to arsenic-induced renal diseases decreased after the improvement of the drinking water supply system through elimination of arsenic exposure from artesian well water. Standardized mortality ratios (SMRs) for renal diseases were calculated for the BFD endemic area for the years 1971-2000. Results show that mortality from renal disease declined gradually after improvement of the drinking-water supply system to eliminate arsenic from artesian well water. Based on the reversibility criterion, the association between arsenic exposure and mortality attributed to renal disease is likely to be positively correlated. PMID- 15799626 TI - Comparison of tissue dosimetry in the mouse following chronic exposure to arsenic compounds. AB - Several chronic bioassays have been conducted in multiple strains of mice in which various concentrations of arsenate or arsenite were administered in the drinking water without a tumorigenic effect. However, one study (Ng et al., 1999) reported a significant increase in tumor incidence in C57Bl/6J mice exposed to arsenic in their drinking water throughout their lifetime, with no tumors reported in controls. A physiologically based pharmacokinetic model for arsenic in the mouse has previously been developed (Gentry et al., 2004) to investigate potential differences in tissue dosimetry of arsenic species across various strains of mice. Initial results indicated no significant differences in blood, liver, or urine dosimetry in B6C3F1 and C57Bl/6 mice for acute or subchronic exposure. The current work was conducted to compare model-predicted estimates of tissue dosimetry to additional kinetic information from the (C57Bl/6 xCBA)F1 and TgAc mouse. The results from the current modeling indicate that the pharmacokinetic parameters derived based on information in the B6C3F1 mouse adequately describe the measured concentrations in the blood/plasma, liver, and urine of both the (C57Bl/6 x CBA)F1 and TgAc mouse, providing further support that the differences in response observed in the chronic bioassays are not related to strain-specific differences in pharmacokinetics. One significant finding was that no increases in skin or lung concentrations of arsenic species in the (C57Bl/6 x CBA)F1 strain were observed following administration of low concentrations (0.2 or 2 mg/U of arsenate in the drinking water, even though differences in response in the skin were reported. These data suggest that pharmacodynamic changes may be observed following exposure to arsenic compounds without an observable change in tissue dosimetry. These results provided further indirect support for the existence of inducible arsenic efflux in these tissues. PMID- 15799628 TI - Effects of maternal versus direct exposure to pulp and paper mill effluent on rainbow trout early life stages. AB - The acute and chronic effects of secondary-treated effluent from a New Zealand pulp and paper mill were assessed using both long-term adult and early life stage (ELS) laboratory exposures of rainbow trout. The relative impact of maternal exposure versus ELS exposure was assessed by a comparison of directly exposed eggs and larvae with the eggs and larvae of exposed adult trout that were reared in reference water. Rainbow trout were exposed to a secondary-treated mixed thermomechanical/bleached kraft mill effluent at a concentration of 15% or to reference water from the egg through to 320-d-old juveniles. The 2 adult rainbow trout exposures were undertaken with nominal concentrations of 10% and 12% treated effluent, respectively. There was no marked effect of water hardening with 15% effluent on fertility or survival of eggs to 16 d. In a subsequent exposure (with hardening in reference water), no significant effects were found on mortality to hatch, time to hatch, length at hatch, mortality to swim-up, mortality to 320 d, or deformity rate at hatch. At experimental termination (320 d), direct-exposed juveniles had smaller livers and reduced condition factor, likely due to differences in food consumption. In 2 subsequent consecutive experiments, exposure of adult trout to 10% and 12% effluent for 2 mo, followed by incubation of the fertilized eggs in reference water, produced no impact on fertility, survival to hatch, survival to swim-up, or length and weight of fry at swim-up. Exposure of adult trout to 12% treated effluent for 8 mo prior to egg fertilization also did not result in differing rates of fertility, mortality to hatch or mortality to swim-up. However, the 8-mo maternal exposure did result in swim-up fry that were significantly shorter and weighed less than the reference swim-up fry. This difference was directly attributable to smaller eggs in the 8 mo-exposed female trout. These results demonstrate that this pulp and paper mill effluent is more likely to elicit indirect impacts on progeny size through chronic exposure of adults to effluent during gonadal recrudescence rather than through direct exposure of early life stages to effluent. PMID- 15799627 TI - Prevention of zearalenone-induced hyperestrogenism in prepubertal mice. AB - Previous methods for the control of zearalenone (ZEN)-induced hyperestrogenism in animals have proven largely ineffective. The main objective in this study was to identify an enterosorbent that decreases the dietary bioavailability, and subsequent estrogenic effects, of ZEN. Initial in vitro screenings in aqueous solution (4 microg ZEN/ml) indicated that an activated carbon (AC) was the most efficient sorbent (99%), followed by a combination of 2 parts AC plus 3 parts HEC (hectorite) (69%), cetylpyridinium-exchanged low-pH montmorillonite (CP-LPHM) clay (58%), hexadecyltrimethylammonium-exchanged low-pH montmorillonite (HDTMA LPHM) clay (54%), and HEC alone (28%). Results from the adult hydra bioassay suggested that the addition of either AC or HEC effectively decreased the effects of ZEN on Hydra attenuata without toxicity, as was observed with the use of either CP-LPHM or HDTMA-LPHM. Based on these results, AC, HEC, and 2AC:3HEC were evaluated in prepubertal mice. At a dietary inclusion level of 0.8% (w/w), AC alone significantly protected mice against the estrogenic effects induced by 35 mg ZEN/kg feed. Inclusion of 1.2% HEC with the 0.8% AC showed no additional protection; whereas 1.2% HEC alone failed to decrease the estrogenic effects. Ground flaxseed (25% w/w) in the diet also elicited protection, but to a lesser extent. Preliminary studies suggested that three similar carbons failed to decrease ZEN bioavailability. These findings suggest that the AC used in this study may be efficacious as an enterosorbent in animals consuming ZEN contaminated diets. However, further studies are needed to evaluate the binding specificity, as well as the safety of chronic exposure. PMID- 15799629 TI - Antioxidant activity of isoacteoside from Clerodendron trichotomum. AB - The antioxidant properties of isoacteoside, isolated from Clerodendron trichotomum (Verbenaceae), were investigated. This compound scavenged intracellular reactive oxygen species (ROS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and prevented lipid peroxidation. This radical scavenging activity of isoacteoside protected cell viability of Chinese hamster lung fibroblast (V79-4) cells exposed to hydrogen peroxide (H2O2). Furthermore, isoacteoside reduced the apoptotic cells formation induced by H2O2, as demonstrated by the decreased number of sub-G1 hypo-diploid cells and apoptotic cell body formation. However, isoacteoside increased the activities of cellular antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT). Taken together, these findings suggest that isoacteoside, isolated from C. trichotomum, possesses antioxidant properties. PMID- 15799630 TI - Anatomy of a dilemma. PMID- 15799631 TI - Research takes a village: 2004 Doris Schwartz award lecture. PMID- 15799632 TI - Nurse staffing in nursing homes in the United States: Part II. AB - This study compared state minimum staffing standards with actual nurse staffing levels in nursing homes in each state. The actual nurse staffing levels in nursing homes were compared to the nurse staffing levels recommended by experts in 2000 and 2001. Data for the study came from primary data collected during a survey of state staffing standards and secondary data of actual staffing federal administrative from records. Actual nurse staffing levels in nursing homes were higher than state minimum standards, showing no evidence that minimum staffing standards become the average staffing level. When actual nurse staffing levels were compared with the minimum standards recommended in a federal report, 97% of all nursing homes had staffing levels below those considered safe. Considerable new federal and state resources are needed to bring the state staffing standards and the actual nursing home staffing levels up to the levels recommended by experts. PMID- 15799633 TI - Assessing pain as a fifth vital sign in long-term care facilities: Recommendations from the field. AB - In long-term care facilities, pain management is complex because dementia, delirium, and other reasons for residents' altered communication ability are a significant barrier to pain assessment. The purpose of this study was to explore the status of implementation of pain as a fifth vital sign in a sample of long term care facilities. A three-round Delphi survey was used to obtain consensus from personnel in 60 long-term care facilities in NY State. Findings are presented in terms of recommendations related to pain criteria, assessment methods, frequency of pain assessment, responsibility for pain assessment, monitoring strategies, education, documentation, and pain management education. The results of this study highlight many important considerations in the treatment of pain as a fifth vital sign in long-term care facilities. Evidence based practice will be facilitated by further research related to underexplored aspects of pain assessment and management, and further attention to care delivery systems that support continued knowledge acquisition and the implementation of best practices. PMID- 15799634 TI - Preventing falls in acute care: an innovative approach. AB - Falls among elderly individuals have been significant sources of disability and death. Falls have affected as many as 10% of older adults during an acute care inpatient stay. The acute care environment has contributed to elderly patient falls. Additionally, manifestations of acute illness, polypharmacy, and medication side effects have been risk factors for falls in the acute care setting. Individualized fall prevention strategies, initial patient assessments, and ongoing patient reassessments have been linked to a decrease in falls in the acute care setting. Approaches to fall prevention have included identification of high-risk patients, communication among staff and family members about an individual's risk of falls, and both case-specific and universal interventions for fall prevention. The purpose of this article is to describe a fall prevention program instituted in an acute care setting in southern Arizona that has produced encouraging results. Moreover, this article addresses individualizing interventions through a continuous clinical feedback loop, which provides patient care areas with relevant information about their patients who fell and recommendations for improving fall prevention. PMID- 15799635 TI - A training program for managing agitation of residents in long-term care facilities: description and preliminary findings. AB - The purpose of this article is to describe a training program for managing agitation of long-term care residents and to report results of a pilot study. The program emphasizes the development of behavioral skills for the assessment, prevention, and reduction of both aggressive and non-aggressive agitated behavior. It includes 8 hours of class instruction followed by 8 hours of weekly supervision by the trainers. The nursing staff of all working shifts of a unit located in a large nursing home and the residents of this unit participated in the pilot study. Residents and staff were assessed prior to and after the 2-month training program. Staff members reported using behavioral techniques to a greater extent and feeling more effective in managing agitation after training. More than 90% of staff members were satisfied with the training program. During the supervision period, the staff developed and implemented individualized interventions for two residents. The interventions involved providing more attention to these residents and, in one case, modifying some aspects of the direct environment which seemed to trigger agitation. Both residents were less agitated after the interventions were implemented. Moreover, there was a reduction in the number and frequency of agitated behaviors for the other residents of the trainees' unit following staff training. PMID- 15799636 TI - Grandparents parenting grandchildren: assessing health status, parental stress, and social supports. AB - This study describes the physical and emotional health of grandparents raising grandchildren and the extent to which the health of this at-risk population is affected further by the caregiving role. A sample of 17 grandparents who are primary caregivers to their grandchildren and reside within a three-state metropolitan area were interviewed in their homes during a 1-year period. Data were collected on physical and mental health status and health problems, level of health, level of stress, economic difficulties, and family and supports. Qualitative data analyses using a panel of experts and NUD*IST, and quantitative methods were used. Results showed wide variation in physical and mental health status; however, most participants tended to score high on parental stress particularly in the child domain. Disruption of social activities, financial problems, and alterations in family relationships were of significant concern. PMID- 15799637 TI - [Women with pelvic complaints and atypical varicose veins, varicose veins of the vulva and insufficiency of the pelvic veins; treatment with embolisation]. AB - In three women, aged 34, 52 and 30 years, respectively, who suffered from chronic pelvic symptoms such as a heavy feeling and abdominal pain, atypical varicose veins were observed on the medial side of the upper thighs together with varicose veins of the vulva; these are indicators of insufficiency of the pelvic veins. The symptoms were reduced following embolisation of the insufficient pelvic veins. These symptoms are also known as the pelvic congestive syndrome. The exact correlation between insufficiency of the pelvic veins and the complaints mentioned above has not yet been determined, but it seems that in most patients the symptoms diminish or disappear following embolisation of the insufficient pelvic veins. When patients complain of chronic pelvic pain of unknown aetiology, one should look for atypical varices and vulval varices and consider pelvic-vein insufficiency as a possible cause. PMID- 15799638 TI - [Improved diagnostics in invasive aspergillosis and systematic monitoring in patients at high risk of infection]. AB - Invasive aspergillosis remains an important cause of morbidity and mortality in patients with prolonged and severe immune suppression such as following haematopoietic stem-cell transplantation. Consensus definitions, which allow categorisation of patients based on diagnostic criteria, are an important improvement in uniform registration of invasive mycoses in clinical trials. Prospective monitoring of high-risk patients for the circulating aspergillus cell wall component galactomannan, results in earlier diagnosis in two-thirds of patients when compared with conventional diagnostic methods. High-resolution CT (HRCT) enables the lesions characteristic of invasive mycoses to be detected earlier and better than by chest radiograph. In addition, invasive mycoses cause characteristic lesions on the HRCT scan including the halo-sign and the air crescent sign. The pre-emptive management strategy which combines monitoring of patients for surrogate markers with a HRCT scan appears to be a promising approach to the early identification and treatment of patients with invasive aspergillosis. PMID- 15799639 TI - [The diagnosis of pulmonary embolism: the best strategies according to the results of a large Dutch multicentre study]. AB - The Dutch national guidelines for the diagnosis of patients with clinically suspected pulmonary embolism (from 1992 en 1998) are poorly followed in clinical practice, due especially to practical objections. A large multicentre trial to investigate the diagnostic accuracy of the available modalities and of recently developed techniques such as D-dimer determination, (99m)Tc-gas-scintigraphy and spiral CT scanning was started in I997. In the first phase, the diagnostic value was assessed of: spiral CT (sensitivity 69%, specificity 84%; sensitivity for segmental and larger pulmonary embolisms (PE) 86% and for subsegmental PE 21%), (99m)Tc-gas-ventilation scintigraphy (no improvement compared to conventional ventilation scintigraphy), D-dimer determination (sensitivity for segmental PE 93% and for subsegmental PE 53%, specificity 63%), clinical decision rules (in combination with D-dimer determination; sensitivity 100%, specificity 11%) and echography of the deep venous system (sensitivity 26% for segmental PE and 7% for subsegmental PE, specificity 97%). In the second phase, the feasibility of two new potentially cost-effective diagnostic algorithms was evaluated on the basis of the results obtained in the first phase and data in the literature. In 631 patients, a clinical risk estimate was made and D-dimer determination was done, followed by a ventilation-perfusion scan and serial compression echography of the leg veins. An apparent recurrence of PE occurred in 6 of 466 patients in whom no PE had been found originally (1.3%; 95% CI: 0.5-2.8). The average costs were 812 Euro,--per patient. In 510 patients, a spiral CT followed by compression echography was performed. Recurrent PE occurred in 3 of 378 patients with initial normal tests (0.8%; 95% CI: 0.2-2.3). The average costs were 883 Euro,--per patient. A combination of both strategies can be cost-effective with a cost 674 Euro,--per patient (recurrence rate: 1.9%). Both the strategy starting with a clinical-risk estimate and a D-dimer determination as well as the strategy consisting of spiral CT and serial echography were safe and cost-effective. According to the results of a survey of hospital directors, internists and pulmonologists, both are well accepted in clinical practice. PMID- 15799641 TI - [Diagnostic image (231). A woman with bluish sweat]. AB - A 56-year-old woman had bluish discoloration of the upper part of white cotton underclothing, caused by apocrine chromhidrosis. PMID- 15799640 TI - [Avoidable visual impairment in The Netherlands: the project "Vision 2020 Netherlands" of the World Health Organization]. AB - In the year 2004 there were an estimated 220,000-320,000 people in The Netherlands with visual impairment. In 150,000-220,000 (70%) of them the visual impairment is either curable or could have been prevented. Those most at risk are people with intellectual disabilities, elderly people in care institutions, elderly people in general and diabetics. 'Vision 2020 Netherlands', an initiative of the World Health Organization, was launched to eliminate avoidable visual impairment in the Netherlands by the year 2020 by means of awareness campaigns, implementation of screening programmes and by expanding eye care capacity through efficient cooperation between the professional groups involved in eye care. PMID- 15799642 TI - [Kidney transplant survival rates and surgical complications in kidney transplants in children; experiences in the Emma Children's Hospital AMC]. AB - OBJECTIVE: To describe surgical complications, and patient and kidney transplant survival rates in children who have had a kidney transplant. DESIGN: Retrospective. METHOD: Data were analysed concerning the children who had been treated with a renal transplantation in the period 1985-2001 because of terminal renal insufficiency in the Emma Children's Hospital of the Academic Medical Centre in Amsterdam, The Netherlands, with arbitrary end date October 7, 2002. RESULTS: In the study period, 55 primary kidney transplantations were performed on 24 girls and 31 boys. 13 living related and 42 post mortem transplantations were performed. The extra-peritoneal approach was performed in all recipients. Up to October 7, 2002, 14 surgical complications (25%) developed: 4 cases of renovascular thrombosis, 4 urinary leakages, 6 urethral strictures, 1 urethral necrosis, 5 haematomas and 3 lymphoceles. 6 patients lost their grafts due to surgical complications (11%); 4 due to thrombosis, 1 due to urethral necrosis and 1 due to haemorrhage or haematoma. Due to hypertension secondary to the underlying kidney disease 2 patients died within one year after transplantation from hypertensive encephalopathy and cerebral bleeding respectively. The one- and five-year graft survival was 83% and 74% for living related transplantations respectively, and for post mortem transplantations 78% and 68% respectively. The main causes of graft loss were chronic rejection (9/21; 43%), acute rejection (4/21; 19%), thrombosis (4/21; 19%) and surgical complications (2/21; 10%). Primary nonfunction was the only factor with a negative prognostic value for graft survival. PMID- 15799643 TI - [Complete atrioventricular septal defect in children with Down's syndrome: good results of surgical correction at younger and younger ages]. AB - OBJECTIVE: To evaluate the results of cardiosurgical treatment of children with Down's syndrome and a complete atrioventricular septal defect (cAVSD). DESIGN: Retrospective. METHOD: Data were collected from the records of all patients with Down's syndrome who had been subjected to primary corrective surgery for cAVSD in the period 1980-2003 in Leiden, The Netherlands. Exclusion criteria were: concomitant tetralogy of Fallot or interruption of the aortic arch and palliative banding of the pulmonary artery. Children with an abnormal shape of the ventricle making it impossible to correct biventricularly were not considered in the study. Data on mortality >30 days after the operation and on repeated surgery were included in the analysis if the duration of follow-up was at least 5 years. RESULTS: The group comprised 148 children: 75 girls and 73 boys. The median age at time of surgery was 20 weeks (range: 6 weeks to 3.7 years) and showed a statistically significant decrease during the period under investigation. Of 4 children lost to clinical follow-up actual data could be obtained. The median duration offollow-up was 6 years and 7 months (range: 38 days to 23 years and 11 months). 28 children (19%) died. The mortality within 30 days after the operation decreased from 0-38% in the period 1980-1989 via 0-30% in the period 1990-1999 to 0% in 2000-2003. The percentage ofreoperations related to the correction ofcAVSD (14%; 14/98) did not seem to increase. Of the correction-related reoperations, 14% (2/14) were followed by a second reoperation. Conclusion. In the period 1980 2003, children with Down's syndrome and a cAVSD were corrected surgically at a younger and younger age. The early mortality decreased to 0% in the years 2000 2003. Echocardiography in the first weeks of life for all children with Down's syndrome makes it possible to diagnose those with a cAVSD early enough for surgical repair. PMID- 15799644 TI - [Allergic bronchopulmonary aspergillosis in asthma and cystic fibrosis]. AB - Three children developed allergic bronchopulmonary aspergillosis (ABPA) as a complication of either asthma or cystic fibrosis (CF). The first patient was a 14 year-old boy with CF who presented with an episode of haemoptysis and a decrease in lung function. He was initially treated with intravenous antibiotics but there was no improvement of his lung function. After starting prednisone-itraconazole his condition improved substantially. The second patient was a 16-year-old girl with CF complicated by ABPA. She was treated for 2 years with prednisone itraconazole. Although the symptoms worsened when the prednisone dosage was gradually reduced, her growth retardation and increased weight decided us to stop prednisone treatment. Two years later, her CF was once again complicated by ABPA. The third patientwas a 16-year-old boy with asthma who had initially been treated for an asthma exacerbation. In retrospect, the cause of his pulmonary exacerbation was probably an ABPA episode. These cases illustrate how important but also how difficult the early diagnosis of ABPA is, and the dilemmas faced in treatment to prevent the fibrotic end stage. PMID- 15799645 TI - [Pneumothorax as a complication of chronic necrotizing pulmonary aspergillosis]. AB - A 65-year-old man with chronic obstructive pulmonary disease who had frequently been treated with prednisone for recurrent exacerbations was admitted to hospital with a pneumothorax. This proved to be a complication of a chronic necrotizing pulmonary aspergillosis. Despite maximal therapy the patient died 15 months after diagnosis from total respiratory insufficiency, the end stage of the bullous emphysema. PMID- 15799646 TI - [Neuroleptic malignant syndrome in users of risperidone]. PMID- 15799648 TI - [Clinical reasoning and decision-making in practice. A 31-year-old woman with transient monocular blindness and polycythaemia]. PMID- 15799647 TI - [Toxicity of dioxins in humans]. PMID- 15799649 TI - Lucky for some. PMID- 15799650 TI - Delayed childbearing--ensuring life choices are informed. PMID- 15799651 TI - Patient perspective. PMID- 15799652 TI - SNAP guide. PMID- 15799653 TI - Management of infertility in Australian general practice. AB - The BEACH program, a continuous national study of general practice activity in Australia, gives us an analysis of the management of infertility between April 1998 and March 2004. Subfertility, which accounts for about 10% of the diagnoses analysed here, is included in the label 'infertility'. Because management rates and methods differ markedly between male and female patients, each gender is analysed separately. This provides a backdrop against which relevant articles in this issue of Australian Family Physician can be further considered. PMID- 15799654 TI - 'We're having trouble conceiving...'. AB - BACKGROUND: The growing trend for Australian women to delay childbearing has a significant impact on their individual fertility and the consequent need for assisted reproductive technology. OBJECTIVE: This article defines infertility and highlights typical male, female, and unexplained combined causes of infertility. Initial fertility investigations best undertaken in general practice are outlined, and suggestions on when to refer a couple to a fertility specialist is clarified. DISCUSSION: Initial assessment of the subfertile couple is best managed by the general practitioner whose relationship can facilitate quick understanding of how long the couple has been trying to conceive, age, and medical history. The woman can then undergo simple global blood and hormone tests while a semen analysis is appropriate for the man. On review, any abnormal tests can indicate referral to a fertility specialist for review and management. PMID- 15799655 TI - Management of the infertile couple. AB - BACKGROUND: With the further development of reproductive technology over the past 2 decades, options for subfertile couples have greatly increased. OBJECTIVE: This article discusses the assessment of common male and female causes of infertility and describes a logical routine for the investigation and treatment of the subfertile couple using a flow chart approach. DISCUSSION: We suggest an approach to assessment for general practitioners, and outline the likely course of more sophisticated testing and treatments that their patients may then experience. PMID- 15799656 TI - The ART of assisted reproductive technology. AB - BACKGROUND: One in six Australian couples of reproductive age experience difficulties in conceiving a child. Once a couple has been appropriately assessed, assisted reproductive technology (ART) techniques can be used to overcome problems with ovulation, tubal patency, male fertility or unexplained infertility. OBJECTIVE: This article discusses the range of ART techniques available to subfertile couples. DISCUSSION: Intracytoplasmic sperm injection is now the most common form of ART used in Australia, being used in almost half of all fresh cycles, with in vitro fertilisation being used in approximately one third of all fresh cycles. PMID- 15799657 TI - Developments in infertility therapy--diagnosis of genetic disease in embryos. AB - BACKGROUND: The general practitioner plays a vital role in assisting couples through the difficult and emotional field of assisted reproductive techniques including in vitro fertilisation (IVF). OBJECTIVE: This article discusses the most recent developments in IVF so that GPs may better explain this ever changing field of medicine to their patients. DISCUSSION: This article begins by explaining how sperm can be brought together with oocytes for fertilisation outside the body, and then moves on to selecting the most genetically appropriate embryos for implantation. From this comes the issue of examining the fertilised oocytes to detect abnormalities, and the options available to couples with ethical concerns. The promise of regenerative medicine through the use of sperm cells is also discussed. PMID- 15799659 TI - Infertility and its treatment--an emotional roller coaster. AB - BACKGROUND: Receiving a diagnosis of infertility is a significant life crisis, and subsequent in vitro fertilisation (IVF) treatment presents a physical and psychological challenge for many patients. OBJECTIVE: This article aims to provide the health professional with an insight into the psychological stressors associated with the experience of infertility, and provide strategies for providing support during this difficult journey. DISCUSSION: Feelings of grief and loss are very common as couples come to terms with the fact that they are not able to conceive naturally. Different emotional coping styles between men and women may add to what is an already stressful time. The grieving process in IVF is often stagnated and chronic, and acceptance and resolution is not psychologically possible until closure is achieved, either by becoming pregnant and giving birth, or ending infertility treatment and ceasing trying to conceive. PMID- 15799658 TI - Polycystic ovary syndrome--a management update. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is the most common endocrinopathy in women and is associated with both reproductive and metabolic disorders. OBJECTIVE: This article discusses the health risks, clinical assessment and investigations consistent with the new internationally agreed definition of PCOS, available treatments, and the long-term monitoring of women with PCOS. DISCUSSION: Women with PCOS have an increased risk of endometrial carcinoma, type 2 diabetes mellitus, and possibly cardiovascular disease. Polycystic ovary syndrome is a diagnosis of exclusion of other causes of hyperandrogenism. Screening for diabetes is important. Treatment is directed at the presenting symptom as the primary cause is unknown. Long-term medical treatment may include the oral contraceptive pill or metformin. Long-term surveillance is recommended for the early detection and treatment of potential metabolic complications. PMID- 15799660 TI - Patient education. Basal body temperature chart. PMID- 15799661 TI - Management of benign breast conditions. Part 1--Painful breasts. AB - This is the first article in a series on breast disorders with an emphasis on diagnosis and management in the general practice setting. This article provides an overview of the investigation of patients with a breast symptom and discusses the assessment and management of mastalgia. PMID- 15799662 TI - Patient education. Breast pain. PMID- 15799663 TI - Recurrent vulvovaginal candidiasis. AB - BACKGROUND: Recurrent vulvovaginal candidiasis affects up to 5% of premenopausal women. It is often associated with significant morbidity and may be difficult to manage. OBJECTIVE: This article discusses the pathogenesis investigations and management of recurrent vulvovaginal candidiasis. DISCUSSION: Recurrent vulvovaginal candidiasis may be misdiagnosed as presenting signs and symptoms are not specific. Examination and microbial testing are required to confirm the diagnosis. Some women appear to have an abnormal host response to the presence of candida species in the vagina, making them susceptible to recurrent episodes of symptomatic infection. Women with recurrent vulvovaginal candidiasis generally respond to a course of suppressive treatment, but many relapse after ceasing therapy. PMID- 15799664 TI - Motor vehicle accidents during episodes of hypoglycaemia--case reports and lessons to be learnt. AB - BACKGROUND: Hypoglycaemia is a common adverse effect of insulin treatment in individuals with diabetes, potentially leading to cognitive impairment, altered levels of consciousness and delayed reactions. If this happens while driving a motor vehicle, a disastrous accident may ensue. OBJECTIVE: This article describes the clinical scenarios of five men with type 1 diabetes who were involved in motor vehicle accidents due to severe hypoglycaemia and discusses the potential risk factors for hypoglycaemia. DISCUSSION: Reference is made to the specific management guidelines set out by the Australian Driver Licensing Authorities for individuals with diabetes. Sometimes, important ethical decisions can be difficult when managing diabetic individuals who fail to comply with therapy or who suffer hypoglycaemia related events and continue to drive a motor vehicle. PMID- 15799665 TI - Implantable hearing devices--beyond hearing aids. PMID- 15799666 TI - Sex, consent and intellectual disability. AB - BACKGROUND: People with intellectual disability, as well as people with dementia or mental health disability, are vulnerable to sexual abuse and may lack the capacity to give valid consent to medical treatment. Their reproductive choices may be limited by their support needs, by social pressures or by sterilisation at a young age. OBJECTIVE: This article uses case vignettes to illustrate key areas in dealing with sexual and reproductive health, and consent issues for people with intellectual disability. It also provides brief guidelines for assessing capacity to consent, and procedures to follow when there is no, or uncertain, capacity. DISCUSSION: People with intellectual disability require education to help them resist abuse, and advocacy to ensure that their environment protects them. They require support and education when making reproductive choices and health care decisions. In cases where they do not have the capacity for informed consent, a substitute decision maker is required. PMID- 15799667 TI - Answering a question about the treatment of croup. PMID- 15799668 TI - The IVF experience--a patient perspective. PMID- 15799669 TI - Failure to diagnose: ectopic pregnancy. AB - Case histories are based on actual medical negligence claims, however certain facts have been omitted or changed by the author to ensure the anonymity of the parties involved. Medical negligence claims alleging 'failure to diagnose' are a common cause of claims and complaints against general practitioners. This article outlines strategies to minimise the possibility of an adverse patient outcome arising out of a delay in diagnosis of ectopic pregnancy. PMID- 15799670 TI - Population and public health in Australian general practice--changes, challenges and opportunities. AB - Public health and medicine have evolved into different disciplines with (at times) competing values and philosophies. The public health role in general practice is expanding. This article reviews the interface between population health, public health, and 'new' public health and general practice, and suggests educational strategies to foster collaboration between these disciplines. PMID- 15799671 TI - Undergraduate general practice attachments--implications and challenges. PMID- 15799672 TI - Reasons for delaying childbearing--a survey of women aged over 35 years seeking assisted reproductive technology. AB - BACKGROUND: Many women seeking assisted reproductive technology for their first planned pregnancy may be unaware of the impact of age on fertility. METHOD: An anonymous mailed survey to women aged over 35 years with no previous planned pregnancies, registered with Monash IVF between January 2002 and May 2003. RESULTS: One hundred and fifty-two out of 266 surveys were completed (57%). The most common reported reasons for delaying pregnancies were: not having a partner (50%), wanting financial security (32%) and a career (19%) before having a family, only recently becoming interested in having children (26%), and being unaware of the impact of age on fertility (18%). DISCUSSION: Some reported reasons for delaying childbearing were beyond personal control. Lack of awareness of the impact of age on fertility may have contributed to the delay in a proportion of the women surveyed. PMID- 15799673 TI - Patient social and economic circumstances--GP perceptions and their influence on management. AB - BACKGROUND: Socioeconomic status (SES) is a major determinant of health. There is little research on if or how general practitioners assess this in individual patients and use it in their clinical practice. METHODS: An exploratory pilot study was conducted using qualitative methodology. Telephone interviews were conducted with a purposive sample of 11 GPs. RESULTS: The participating GPs commonly assessed SES through observation of patient behaviour and presentation, contextualised with knowledge of the patient's background and the community, rather than direct enquiry. The GPs understood the link between low SES and poor health primarily in terms of a higher prevalence of behavioural risk factors. Participating GPs were generally pessimistic about achieving behaviour change for patients they identified as being of low SES. DISCUSSION: General practitioners' complex understanding of SES is supported by newer theories of health inequalities and social position. However, GP attitudes may potentially play a role in reinforcing inequalities. Further research is needed regarding the relationship between GP perceptions and objective indicators of SES. PMID- 15799674 TI - Childhood asthma diagnosis and use of asthma medication. AB - AIM: To determine the burden of asthma in children. METHODS: A cross sectional, randomised, computer assisted telephone survey of a community based sample of 2020 children aged 5-14 years in western Sydney (New South Wales) over a 20 day period from June 2000 to July 2000. RESULTS: Main outcome measures were carer reported history of asthma diagnosis, hospital presentation/admission for asthma, recent use of anti-asthma medications, and recent respiratory symptoms. Diagnosed asthma was reported in 31% (of whom 42% were diagnosed aged 2 years or under) and asthma medications used in the previous year by 21% of children. Factors significantly associated with a reported asthma diagnosis included: male gender (OR: 1.51), birth in Australia (OR: 1.64), living in an English speaking household (OR: 1.47), Aboriginality (OR: 2.32), possession of a health care card (OR: 1.28), previous pneumonia (OR: 2.4) or pertussis (OR: 2.0), and a recent episode of croup (OR: 1.9). Exposure to tobacco smoke and immunisation status were not significant. DISCUSSION: We confirm a high prevalence of asthma and medication use for asthma. The high proportion of children diagnosed asthmatic at 2 years or under (when asthma cannot be diagnosed reliably) suggests overdiagnosis of asthma may contribute to the apparent high prevalence. PMID- 15799675 TI - Self management beliefs--attitudes and behaviour of adults with severe life threatening asthma requiring an admission to hospital. AB - BACKGROUND: Adults who have had an admission for severe life threatening asthma are at high risk of future attacks or death. Optimal self management is the key to reducing this risk but relies on people recognising the severity of symptoms and engaging in specific health care behaviour. METHODS: We used a focus group to examine self management beliefs, attitudes and behaviours in five adults admitted to hospital for asthma. RESULTS: There were a number of themes. Patients delayed seeking medical attention until asthma symptoms were severe despite ownership of a peak expiratory flow meter, written plan, or experience of previous attack. DISCUSSION: Insight into the significance of the severity of symptoms and need to change self management behaviour in response to symptoms is key to reducing the risk of future attacks. The general practitioner has a key role in determining and addressing self management behaviour and attitudes that place patients at risk. PMID- 15799676 TI - Barriers to the use of spirometry in general practice. AB - BACKGROUND: Guidelines advise chronic obstructive pulmonary disease (COPD) should be diagnosed and managed by using spirometry to demonstrate irreversible airflow limitation and monitor change in smokers and ex-smokers aged over 35 years. METHODS: A cross-sectional study of patients and their general practitioners investigating use of spirometry in COPD in two practices by lung function assessment, review of practice records, interviews and focus groups. RESULTS: Sixteen GPs, and 38 patients with a diagnosis of COPD participated. At diagnosis, although 72% had spirometry, this occurred in only 41% of 17 patients diagnosed by a GP; but in all 19 cases when a specialist was involved. Diagnosis often occurred late, despite all patients having previously recorded symptoms typical of COPD. General practitioners expressed a preference to diagnose COPD on clinical grounds. Although 58% of patients had recent spirometry for current management, only 32% were performed by their GP. There were organisational and technical barriers to spirometry and poor recognition of the essential role of spirometry in the diagnosis of COPD. DISCUSSION: There are a number of potentially reversible factors that hinder practice recommendations regarding the use of spirometry in general practice to diagnose and manage COPD. PMID- 15799677 TI - Becoming a GP--a qualitative study of the career interests of medical students. AB - AIM: Only a minority of Australian graduates are interested in careers in general practice. The factors influencing medical students toward general practice as a career choice are poorly understood, even though this is important to the makeup of the medical workforce. METHODS: We ran 10 focus groups involving 82 first and final year medical students from three Australian medical schools in 2002, and analysed factors influencing participants' interest in working in general practice. RESULTS: About half the students were interested in general practice. Attractive factors were: the nature of the work (including its diversity), continuity of care, community context, and working conditions (including flexibility of training and work, availability of part time work and portability of qualifications). Negative factors included: the breadth of knowledge needed, boring work (in urban general practice), having to run a business, and working conditions (including relatively poor remuneration, overwork in rural general practice, and poor status of general practitioners). Some students were strongly influenced by negative attitudes of the GPs they were taught by, deciding against general practice as a career. DISCUSSION: Medical educators and GPs should be aware of this important influence. PMID- 15799678 TI - Safety and efficacy of topical calcineurin inhibitors in the treatment of childhood atopic dermatitis. AB - Approximately 10-20% of infants in industrialized countries experience atopic dermatitis. In recent decades topical corticosteroids have been the first-choice therapy for treatment of flares. However, this form of therapy may induce skin atrophy, especially after application to facial lesions or with long-term use. Thus, development of new anti-inflammatory topical agents for the treatment of childhood atopic dermatitis was needed. The topical calcineurin inhibitors tacrolimus and pimecrolimus have an effect on various cells of the cutaneous immune system, specifically on T cells, by inhibiting the phosphatase calcineurin and preventing the transcription of proinflammatory cytokines. In several clinical studies of children and adults with atopic dermatitis, topical calcineurin inhibitors were found to be effective both on the face and the trunk and extremities, in both short- and long-term treatment regimens. Tachyphylaxis or rebound were not observed. In most patients an improvement of their eczema occurred during the first week of treatment, as measured by subjective and objective clinical signs of atopic dermatitis. Treatment significantly reduced the incidence of flares and the need for corticosteroids in children and adults. Treatment success, commonly defined as 'excellent improvement' or 'clearing of all lesions', was observed in more than one-third of all children treated with 0.03% or 0.1% tacrolimus or 1% pimecrolimus. Topical application of pimecrolimus and tacrolimus does not lead to significant blood concentrations of these agents in the majority of children with atopic dermatitis, and any increase in blood concentrations decreases after a few days of therapy. No changes in laboratory parameters were observed in short- and long-term studies in patients with atopic dermatitis. The most common adverse effect following the application of topical calcineurin inhibitors is mild to moderate symptoms of irritation such as burning, erythema and pruritus, which occurred in up to 20% of all children treated with tacrolimus and 10% of children treated with pimecrolimus, and usually faded after a few days. In contrast to topical corticosteroids, calcineurin inhibitors do not induce skin atrophy, even after long-term use. Topical calcineurin inhibitors have been proven to be effective and have a good safety profile during short-term and long-term use for up to 1 year with pimecrolimus and up to 4 years with tacrolimus. Given the lack of extensive experience with use of topical calcineurin inhibitors over longer periods, regular use of these agents, particularly in children, should be undertaken only after careful consideration of individual cases. Sun protection should also be advised. PMID- 15799679 TI - Cutaneous adverse reactions following anti-infective vaccinations. AB - Although widely administered, anti-infective vaccinations are rarely responsible for cutaneous adverse effects. In this context, hepatitis B and bacillus Calmette Guerin vaccines are the most frequently incriminated products. Cutaneous adverse effects are less frequently encountered following administration of vaccines against varicella, diphtheria/tetanus/pertussis (primary and booster doses), measles, poliomyelitis, rubella, pneumococcus, tick-borne encephalitis, smallpox, Meningococcus and influenza. The adverse effects can occur at the site of or at a distance from the injection. The patho-mechanisms of local adverse cutaneous reactions include predominantly nonspecific lymphoid or granulomatous reactions. Allergic reactions to the vaccine strain, adjuvants, conservatives or other components are less frequently involved in local vaccine adverse effects. Systemic reactions are mainly mediated by immediate type or immune complex related allergic reactions to toxoid-, ovalbumin-, gelatin- or pneumococcal containing vaccines. Systemic reactions are sometimes related to a specific vaccine strain. Other cutaneous reactions may also occur through unknown patho mechanisms. No vaccine type or strain is specifically associated with a particular type of cutaneous adverse effect. This article presents seven case reports of cutaneous adverse effects following anti-infective vaccination then reviews the relevant literature on this subject. PMID- 15799680 TI - Microdermabrasion. AB - Microdermabrasion is a simple, safe, office cosmetic procedure in which aluminum oxide crystals or other abrasive substances are blown onto the face, then vacuumed off, using a single handpiece. This procedure has rapidly become widely utilized for a variety of cosmetic objectives, including the improvement of photoaging, hyperpigmentation, acne, scars and stretch marks. Despite its widespread use, little is known about its actual mechanism of action. The few published studies suggest that patients and physicians alike report a mild benefit when microdermabrasion is utilized for photoaging. Histologic evaluation reveals little actual abrasion of the skin with the procedure, yet changes are seen in the dermis. Given the safety, simplicity and patient satisfaction associated with microdermabrasion, it is likely to remain a popular treatment. PMID- 15799681 TI - Cicatricial pemphigoid (mucous membrane pemphigoid): current and emerging therapeutic approaches. AB - The treatment of cicatricial pemphigoid, also called mucous membrane pemphigoid (MMP), poses a great challenge, because the condition often takes an intransigent course despite all therapeutic efforts. Because of its diverse clinical manifestations, patients with MMP often have to be treated by a variety of specialists, including dermatologists, ophthalmologists, ear, nose, and throat specialists, and dentists. Since there are almost no randomized, controlled, double-blind studies comparing the use of various therapeutic agents in this condition, treatment decisions still rely heavily on individual clinicians' experience. Many different therapeutic regimens have been described in the literature, but only a few seem to hold up as valid alternatives. Systemic corticosteroids are still the agent of first choice, especially as rescue medication, for curtailing acute exacerbations. However, because of their well known long-term adverse effects, corticosteroids must be combined with immunosuppressive and/or anti-inflammatory agents. To determine which drug to choose, it is helpful to categorize patients -- as recommended by the First International Consensus -- in terms of high- and low-risk depending on the site and severity of their disease and on how rapidly it progresses. The recommended treatment for high-risk patients (i.e. patients with ocular, genital, laryngeal, esophageal or nasopharyngeal involvement) is a combination of prednisone and cyclophosphamide, or alternatively azathioprine. Once clinical improvement is evident, the corticosteroids should be slowly tapered. Dapsone is another alternative that may be used in high-risk patients, but patients who do not show any short-term improvement on this regimen should be switched to cyclophosphamide. Intravenous immunoglobulins are another effective, but expensive, treatment option in high-risk patients. Low-risk patients may well be managed with topical therapy alone, such as corticosteroids or cyclosporine. Other systemic options include dapsone, tetracycline, and nicotinamide as well as azathioprine in combination with low doses of corticosteroids. Various other systemic and topical agents, and recently biologics such as etanercept, have been reported to be effective in the treatment of MMP. However, most of the reported cases consisted of only small patient numbers and the true benefit of such agents in the condition is therefore not yet clear. PMID- 15799682 TI - Lack of drug interaction conformity in commonly used drug compendia for selected at-risk dermatologic drugs. AB - BACKGROUND: Systemic treatments that have significant efficacy for dermatologic conditions can often cause severe adverse reactions when improperly combined with interacting drugs. We have noted discrepancies in the reporting of such drug-drug interactions among major drug compendia used in hospitals and physicians' offices. OBJECTIVE: The aim of this study was to compare the consistency of drug drug interaction listings among four of the most widely used English language drug compendia in the US. METHODS: Drug monographs from Mosby's GenRx, USP DI, AHFS Drug Information, and the Physicians' Desk Reference were compared. The respective drug-drug interactions provided by the various compendia for four systemic agents -- dapsone, erythromycin, methotrexate and prednisone -- commonly prescribed for dermatologic purposes were compiled. RESULTS: We found considerable discrepancies among the compendia with respect to the number of drug interactions listed. Agreement among the compendia was especially poor when more than two sources were compared. CONCLUSIONS: There is a need for a drug compendium that reconciles drug-drug interaction reporting discrepancies under standardized criteria based on the scientific literature and clinical significance. PMID- 15799683 TI - Efalizumab. AB - Efalizumab is a humanized monoclonal antibody that binds to CD11a, the alpha subunit of lymphocyte function-associated antigen-1, and consequently inhibits T cell activation. In randomized, double-blind, placebo-controlled trials, efalizumab 1.0 mg/kg, administered subcutaneously once weekly for 12 weeks, significantly reduced disease activity in patients with chronic, moderate-to severe plaque psoriasis. Significantly more efalizumab recipients had a > or =75% decrease in the Psoriasis Area and Severity Index (PASI) score [22.4-38.9%] than placebo recipients (2.4-4.9%); an additional 12 weeks of treatment resulted in sustained or increased PASI responses. The efficacy of weekly subcutaneous efalizumab was maintained during 15 months of treatment. Efalizumab significantly improved health-related quality of life in patients with chronic plaque psoriasis, with significant improvements in all the Dermatology Life Quality Index domains. Efalizumab was generally well tolerated in patients with chronic, moderate-to-severe plaque psoriasis, with few serious adverse events or treatment withdrawals. The most common adverse events were headache, chills, myalgia, pain, and fever; these most often occurred within 2 days of administration of the drug, were most frequent after the first or second dose, and decreased in frequency over time. PMID- 15799687 TI - Etanercept: a review of its use in the management of plaque psoriasis and psoriatic arthritis. AB - Etanercept (Enbrel), a tumor necrosis factor-alpha antagonist produced by recombinant technology, is approved for use in the US as subcutaneous monotherapy in adults with moderate-to-severe psoriasis who are candidates for systemic therapy or phototherapy. The drug is also indicated in patients with psoriatic arthritis, in whom it may be used in combination with methotrexate. In well designed trials in patients with moderate-to-severe psoriasis, short-term etanercept therapy (typically 25 or 50 mg twice weekly) significantly increased the proportion of patients achieving a 75% reduction in the Psoriasis Area and Severity Index score compared with placebo. Similarly, in well designed trials in patients with psoriatic arthritis, treatment with short-term etanercept 25 mg twice weekly, alone or in combination with methotrexate, improved clinical features of the disease, while radiographic progression of joint damage appeared to be significantly slowed in a nonblind 1-year extension. Short-term etanercept therapy was well tolerated in patients with psoriasis or psoriatic arthritis. Etanercept is thus a valuable new option for the treatment of patients with chronic moderate-to-severe plaque psoriasis (who are candidates for systemic therapy or phototherapy or have failed other systemic therapies) or with psoriatic arthritis. PMID- 15799689 TI - Effects of N1, N13-diethylnorspermine (DENSPM) and X-radiation treatment on human colorectal tumor clones with varying X-radiation and drug responses. AB - This study was designed to examine the effects of treatment with N1, N13 diethylnorspermine (DENSPM), a spermine analog, and X radiation on survival and on the polyamine and spermidine/spermine N1-acetyltransferase (SSAT) levels in closely related human colorectal tumor (HCT116) clones exhibiting a wide range of X-radiation and drug responses. After treatment with DENSPM and X radiation, clonogenic cell survival was measured. SSAT protein levels were measured by Western blot analysis and SSAT enzymatic activities by the conversion of [1 14C]acetyl-CoA into [1-14C]acetylspermidine. Polyamine [i.e. putrescine (PUT), spermine (SPM) and spermidine (SPD)] levels were measured with high-performance liquid chromatography. DENSPM enhanced the efficacy of radiation treatment in HCT116, HCT116-Clone2 (a radiation-resistant clone) and HCT116-Clone10 (a clone with similar X-radiation response as the parental HCT116 cells) but not in HCT116 CloneK (an X-radiation-sensitive but relatively drug-resistant clone). Treatment with DENSPM without X radiation caused the most significant increase in SSAT activity (approximately 22-fold) and an almost complete depletion of SPD levels in HCT116-CloneK. Our results suggest that (a) the lack of sensitization of X radiation treatment by DENSPM in HCT116-CloneK was likely due to the prior depletion of SPD levels by DENSPM alone, (b) natural polyamine contents and/or inducibility of SSAT may be important factors influencing cellular response to combined X-radiation and DENSPM treatments, and (c) more importantly, there may be a potentially novel role for combining polyamine analogs such as DENSPM with X rays. PMID- 15799690 TI - Detection of oxidative stress induced by low- and high-linear energy transfer radiation in cultured human epithelial cells. AB - A standardized dichlorofluorescin (DCF) fluorometric assay capable of measuring radiation-induced oxidative stress was used to determine the effectiveness of protons and high-mass, high-atomic number (Z) and high-energy (HZE) particles to produce oxidative stress in vitro. Protons were found to be about equally as effective as X rays in the generation of oxidative stress in cultured cells. However, 56Fe-ion beams with energies of 1 GeV/nucleon and 5 GeV/nucleon were less effective than X rays or gamma rays in inducing dichlorofluorescin (DCFH) oxidation. The relatively lower slope values for the dose responses of HZE particle radiation-induced DCFH oxidation indicate that the sensitivity of the DCF fluorometric assay is probably dependent on the linear energy transfer (LET) of the radiation beam. PMID- 15799691 TI - DNA-incorporated 125I induces more than one double-strand break per decay in mammalian cells. AB - The Auger-electron emitter 125I releases cascades of 20 electrons per decay that deposit a great amount of local energy, and for DNA-incorporated 125I, approximately one DNA double-strand break (DSB) is produced close to the decay site. To investigate the potential of 125I to induce additional DSBs within adjacent chromatin structures in mammalian cells, we applied DNA fragment-size analysis based on pulsed-field gel electrophoresis (PFGE) of hamster V79-379A cells exposed to DNA-incorporated 125IdU. After accumulation of decays at -70 degrees C in the presence of 10% DMSO, there was a non-random distribution of DNA fragments with an excess of fragments <0.5 Mbp and the measured yield was 1.6 DSBs/decay. However, since these experiments were performed under high scavenging conditions (DMSO) that reduce indirect effects, the yield in cells exposed to 125IdU under physiological conditions would most likely be even higher. In contrast, using a conventional low-resolution assay without measurement of smaller DNA fragments, the yield was close to one DSB/decay. We conclude that a large fraction of the DSBs induced by DNA-incorporated 125I are nonrandomly distributed and that significantly more than one DSB/decay is induced in an intact cell. Thus, in addition to DSBs produced close to the decay site, DSBs may also be induced within neighboring chromatin fibers, releasing smaller DNA fragments that are not detected by conventional DSB assays. PMID- 15799692 TI - Alterations of DNA copy number and expression in genes involved in cell cycle regulation and apoptosis signal pathways in gamma-radiation-sensitive SX9 cells and -resistant SR-1 cells. AB - In the present study, genomic differences related to sensitivity to radiation were examined by comparative genomic hybridization and GeneChip 45K microarray in SX9 cells (radiation-sensitive) and their parental line, SR-1 (radiation resistant). SX9 cells have defective DNA-dependent protein kinase catalytic subunit (DNA-PKcs) activity. DNA-PKcs is a DNA double-strand break repair protein that maintains chromosomal stability through nonhomologous end joining. However, the molecular basis of the radiation sensitivity of SX9 cells is unclear. Flow cytometry analysis showed that SR-1 and SX9 cells had a larger G2/M-phase population at 12 h after 4 Gy gamma irradiation, while only SR-1 cells progressed to G1/S at 24-36 h. SX9 and SR-1 cells had similar patterns of DNA copy number alteration, but the gains were observed on chromosome 9 (cent-E2), 11 (cent-A3), and 12 (C1-E) only in SX9 cells. Expression of genes located on those regions is higher in SX-9 cells than in SR1 cells, and the regions include genes associated with apoptosis and cell cycle regulation. Time-course data for gene expression at 0, 1, 3, 6 and 12 h after 4 Gy gamma irradiation revealed that the genes whose expression was altered in SX9 cells but not in SR-1 cells are in 16 clusters. Three of these clusters included genes for cell cycle regulation: JNK, PKC (PRKC) and ceramide cascade protein. These results suggest that amplification and altered expression of genes associated with cell cycle and apoptosis regulators in DNA-PK-deficient SX9 cells affect the differences in response to gamma radiation between SX9 and SR-1 cells. PMID- 15799693 TI - Medium from irradiated cells induces dose-dependent mitochondrial changes and BCL2 responses in unirradiated human keratinocytes. AB - Exposure of unirradiated human keratinocytes to irradiated cell conditioned medium (ICCM) is known to cause a cascade of events that leads to reproductive death and apoptosis. This study investigates the effect of ICCM on clonogenic survival, mitochondrial mass and BCL2 expression in unirradiated keratinocytes. Exposure to 5 mGy, 0.5 Gy and 5 Gy ICCM resulted in a significant decrease in clonogenic survival. Human keratinocytes incubated with ICCM containing an antioxidant, N-acetylcysteine, showed no significant decrease in clonogenic survival. HPV-G cells incubated with ICCM containing a caspase 9 inhibitor showed no significant decrease in clonogenic survival when the ICCM dose was < or =0.5 Gy. A significant increase in mitochondrial mass per cell was observed after exposure to 5 mGy and 0.5 Gy ICCM. A change in the distribution of the mitochondria from a diffuse cytoplasmic distribution to a more densely concentrated perinuclear distribution was also observed at these doses. No significant increase in mitochondrial mass or change in distribution of the mitochondria was found for 5 Gy ICCM. Low BCL2 expression was observed in HPV-G cells exposed to 5 mGy or 0.5 Gy ICCM, whereas a large significant increase in BCL2 expression was observed in cells exposed to 5 Gy ICCM. This study has shown that low-dose irradiation can cause cells to produce medium-borne signals that can cause mitochondrial changes and the induction of BCL2 expression in unirradiated HPV-G cells. The dose dependence of the mitochondrial changes and BCL2 expression suggests that the mechanisms may be aimed at control of response to radiation at the population level through signaling pathways. PMID- 15799694 TI - Genetic factors influencing bystander signaling in murine bladder epithelium after low-dose irradiation in vivo. AB - Radiation-induced bystander effects occur in cells that are not directly hit by radiation tracks but that receive signals from hit cells. They are well documented in vitro consequences of low-dose exposure, but their relevance to in vivo radiobiology is not established. To investigate the in vivo production of bystander signals, bladder explants were established from two strains of mice known to differ significantly in both short-term and long-term radiation responses. These were investigated for the ability of 0.5 Gy total-body irradiation in vivo to induce production of bystander signals in bladder epithelium. The studies demonstrate that irradiated C57BL/6 mice, but not CBA/Ca mice, produce bystander signals that induce apoptosis and reduce clonogenic survival in reporter HPV-G-transfected keratinocytes. Transfer of medium from explants established from irradiated animals to explants established from unirradiated animals confirmed these differences in bladder epithelium. The responses to the in vivo-generated bystander signal exhibit genotypic differences in calcium signaling and also in signaling pathways indicative of a major role for the balance of pro-apoptosis and anti-apoptosis proteins in determining the overall response. The results clearly demonstrate the in vivo induction of bystander signals that are strongly influenced by genetic factors and have implications for radiation protection, medical imaging, and radiotherapy. PMID- 15799696 TI - Semiquantitative detection of cytokine messages in X-irradiated and regenerating rat thymus. AB - We investigated the expression of cytokine mRNA derived from thymocytes or thymic epithelial cells in X-irradiated (8 Gy) and recovering rat thymuses, according to our previous observation (Mizutani et al., Radiat. Res. 157, 281-289, 2002). The changes in mRNA expression level of interleukin 2 (Il2), Il4, tumor necrosis factor alpha (Tnf), interferon gamma (Ifng), and transforming growth factor beta (Tgfb) were examined. The mRNA expression of Il2 and Il4 decreased from day 5 to day 14 after irradiation. Thereafter, the expression level of Il2 mRNA recovered to normal control levels; however, the expression of Il4 mRNA tended toward significantly low levels. Tnf mRNA expression decreased on day 5 after irradiation and then showed a gradual increase back to normal control levels. Tgfb mRNA expression did not change significantly. Ifng mRNA expression was transiently enhanced from day 11 to day 14. The mRNA expression levels of Il10 increased significantly from day 3 to day 7 after irradiation. In addition, the mRNA expression of thymic epithelial cell-derived Il7 showed a transient decrease on day 3; however, then it showed a continuous increase from day 5 to day 21, finally reaching twice the normal control levels after X irradiation. These observations suggest that the expression of cytokine messages in the irradiated thymus changed significantly and did not return to normal for a long time after 8 Gy irradiation. PMID- 15799697 TI - Radiation-induced increase in plasma Flt3 ligand concentration in mice: evidence for the implication of several cell types. AB - Circulating T lymphocytes were proposed as the main producer of Flt3 ligand. However, during aplasia, there is a drastic reduction in the number of T lymphocytes, while plasma Flt3 ligand concentration is increased. This contradiction prompted us to compare variations in plasma Flt3 ligand during radiation-induced aplasia in BALB/c mice and in T-lymphocyte-deficient NOD-SCID mice to delineate the role of T lymphocytes in the increase in Flt3 ligand concentration. The results showed that plasma Flt3 ligand concentration was increased similarly in the two strains of mice, and that Flt3 ligand concentration was negatively correlated to the number of residual hematopoietic progenitors. Moreover, the Flt3 ligand mRNA expression and Flt3 ligand protein concentration were similar in the two strains of mice in all organs tested, i.e. thymus, spleen, bone marrow, liver, brain and blood cells. These results confirm that Flt3 ligand concentration in the blood is a reflection of bone marrow function and that T lymphocytes are not the main regulator of Flt3 ligand variations during aplasia. PMID- 15799695 TI - Intravascular immune surveillance by CXCR6+ NKT cells patrolling liver sinusoids. AB - We examined the in vivo behavior of liver natural killer T cells (NKT cells) by intravital fluorescence microscopic imaging of mice in which a green fluorescent protein cDNA was used to replace the gene encoding the chemokine receptor CXCR6. NKT cells, which account for most CXCR6(+) cells in liver, were found to crawl within hepatic sinusoids at 10-20 microm/min and to stop upon T cell antigen receptor activation. CXCR6-deficient mice exhibited a selective and severe reduction of CD1d-reactive NKT cells in the liver and decreased susceptibility to T-cell-dependent hepatitis. CXCL16, the cell surface ligand for CXCR6, is expressed on sinusoidal endothelial cells, and CXCR6 deficiency resulted in reduced survival, but not in altered speed or pattern of patrolling of NKT cells. Thus, NKT cells patrol liver sinusoids to provide intravascular immune surveillance, and CXCR6 contributes to liver-based immune responses by regulating their abundance. PMID- 15799698 TI - Further study of prolongation of life span associated with immunological modification by chronic low-dose-rate irradiation in MRL-lpr/lpr mice: effects of whole-life irradiation. AB - MRL-lpr/lpr mice carry a deletion in the apoptosis-regulating Fas gene that markedly shortens life due to multiple severe diseases. In our previous study (Radiat. Res. 161, 168- 173, 2004), chronic low-dose-rate gamma irradiation of mice at 0.35 or 1.2 mGy/h for 5 weeks markedly prolonged the life span, accompanied by immunological activation. This report shows that extension of the irradiation period to the entire life of the mice at the same dose rates improved survival further. The 50% survival time for untreated mice, 134 days, was prolonged to 502 days by 1.2 mGy/h life-long irradiation. Also obtained were a time course and a radiation dose-rate response for the activation of the immune system as indicated by a significant increase in CD4+ CD8+ T cells in the thymus and CD8+ T cells in the spleen and also by a significant decrease in CD3+ CD45R/B220+ cells and CD45R/B220+ CD40+ cells in the spleen. Drastic ameliorations of multiple severe diseases, i.e. total-body lymphadenopathy, splenomegaly and serious autoimmune diseases including proteinuria, and kidney and brain-central nervous system syndromes, were found in parallel with these immunological activations, with lifelong low-dose-rate irradiation being more effective than 5-week irradiation at low dose rates. PMID- 15799699 TI - Long-term follow-up for brain tumor development after childhood exposure to ionizing radiation for tinea capitis. AB - Ionizing radiation is an established risk factor for brain tumors, yet quantitative information on the long-term risk of different types of brain tumors is sparse. Our aims were to assess the risk of radiation-induced malignant brain tumors and benign meningiomas after childhood exposure and to investigate the role of potential modifiers of that risk. The study population included 10,834 individuals who were treated for tinea capitis with X rays in the 1950s and two matched nonirradiated groups, comprising population and sibling comparison groups. The mean estimated radiation dose to the brain was 1.5 Gy. Survival analysis using Poisson regression was performed to estimate the excess relative and absolute risks (ERR, EAR) for brain tumors. After a median follow-up of 40 years, an ERR/Gy of 4.63 and 1.98 (95% CI = 2.43-9.12 and 0.73-4.69) and an EAR/Gy per 10(4) PY of 0.48 and 0.31 (95% CI = 0.28-0.73 and 0.12-0.53) were observed for benign meningiomas and malignant brain tumors, respectively. The risk of both types of tumors was positively associated with dose. The estimated ERR/Gy for malignant brain tumors decreased with increasing age at irradiation from 3.56 to 0.47 (P = 0.037), while no trend with age was seen for benign meningiomas. The ERR for both types of tumor remains elevated at 30-plus years after exposure. PMID- 15799700 TI - Radiation abolishes inducer binding to lactose repressor. AB - The lactose operon functions under the control of the repressor-operator system. Binding of the repressor to the operator prevents the expression of the structural genes. This interaction can be destroyed by the binding of an inducer to the repressor. If ionizing radiations damage the partners, a dramatic dysfunction of the regulation system may be expected. We showed previously that gamma irradiation hinders repressor-operator binding through protein damage. Here we show that irradiation of the repressor abolishes the binding of the gratuitous inducer isopropyl-1-beta-D-thiogalactoside (IPTG) to the repressor. The observed lack of release of the repressor from the complex results from the loss of the ability of the inducer to bind to the repressor due to the destruction of the IPTG binding site. Fluorescence measurements show that both tryptophan residues located in or near the IPTG binding site are damaged. Since tryptophan damage is strongly correlated with the loss of IPTG binding ability, we conclude that it plays a critical role in the effect. A model was built that takes into account the kinetic analysis of damage production and the observed protection of its binding site by IPTG. This model satisfactorily accounts for the experimental results and allows us to understand the radiation-induced effects. PMID- 15799701 TI - Track structure in DNA irradiated with heavy ions. AB - The spatial properties of trapped radicals produced in heavy-ion-irradiated solid DNA at 77 K have been probed using pulsed electron paramagnetic double resonance (PELDOR or DEER) techniques. Salmon testes DNA hydrated to 12 water molecules per nucleotide was irradiated with 40Ar ions of energy 100 MeV/nucleon and LET ranging from 300 to 400 keV/microm. Irradiated samples were maintained at cryogenic temperature at all times. PELDOR measurements were made using a refocused echo detection sequence that allows dipolar interaction between trapped radicals to be observed. The EPR spectrum is attributed to electron loss/gain DNA base radicals and neutral carbon-centered radicals that likely arise from sugar damage. We find a radical concentration of 13.5 x 10(18) cm(-3) in the tracks and a track radius of 6.79 nm. The cross section of these tracks is 144 nm2, yielding a lineal radical density of 2.6 radicals/nm. Based on the yields determined previously for particles having calculated LET values of 300-400 keV/microm and our measured lineal density, we obtain an LET of 270 keV/microm, which is in good agreement with the calculated range of values. These measurements of radical density and spatial extent provide the first direct experimental determination of track characteristics in irradiated DNA. PMID- 15799702 TI - Yield of OH radicals in water under high-density energy deposition by heavy-ion irradiation. AB - The purpose of the present study was to evaluate the dependence of the OH radical yield on the atomic number and the energy of the heavy ions to understand chemical reactions of aqueous solutions. The total yields of oxidized products from phenol in water increased superlinearly as the incident energy increased from 5 MeV/nucleon to 18 MeV/nucleon for carbon and neon ions. The radiolytic yields of OH radicals produced by the ions were determined by analyzing the relationships of the oxidation yields of phenol to the incident energies up to 18 MeV/nucleon for ions in the range of LET from 110 eV/nm to 550 eV/nm and from 320 eV/nm to 1100 eV/nm for carbon and neon ions, respectively. The yields of the OH radicals increased with the specific energy for the same kind of ion and decreased with the atomic number for different ions used at the same specific energy. PMID- 15799703 TI - Assessment of 90Sr concentration in dental tissue using thin-layer beta-particle detectors and verification with numerical calculations. AB - Electron paramagnetic resonance (EPR) measurements of tooth enamel can be used as an individual biological dosimeter for external dose assessment. However, the presence of 90Sr in the tooth tissues makes the task of interpreting EPR tooth dosimetry more complicated. The determination of the dose contribution of incorporated 90Sr in calcified tissue to the total dose measured by EPR is one of the main aspects of correct interpretation of EPR tooth dosimetry. In this work, experimental and numerical calculations were performed to convert the measured beta-particle dose rate to 90Sr concentration in calcified tissue. The cumulative beta-particle dose was measured by exposing artificially contaminated dentin and enamel to thin-layer alpha-Al2O3:C detectors in two different exposure geometries. Numerical calculations were performed for experimental exposure conditions using calculations of electron transport and secondary photons [Monte Carlo n-Particle Transport code version 4C2 (MCNP)]. Numerical calculations were performed to optimize the sample size and exposure geometry. The applicability of two different exposure conditions to be used in routine analysis was tested. Comparison of the computational and experimental results demonstrated very good agreement. PMID- 15799704 TI - Monte Carlo simulation of the spatial distribution of energy deposition for an electron microbeam. AB - Dosimetry calculations characterizing the spatial variation of the energy deposited by the slowing and stopping of energetic electrons are reported and compared with experimental measurements from an electron microbeam facility. The computations involve event-by-event, detailed-histories Monte Carlo simulations of low-energy electrons interacting in water vapor. Simulations of electron tracks with starting energies from 30 to 80 keV are used to determine energy deposition distributions in thin cylindrical rings as a function of penetration and radial distance from a beam source. Experimental measurements of the spatial distribution of an electron microbeam in air show general agreement with the density-scaled simulation results for water vapor at these energies, yielding increased confidence in the predictions of Monte Carlo track-structure simulations for applications of the microbeam as a single-cell irradiator. PMID- 15799705 TI - Radiation-induced genomic instability and bystander effects; implications for evolutionary biology. PMID- 15799706 TI - Curvature in the dose response of the life span study cancer mortality data: comments on "effect of recent changes in atomic bomb survivor dosimetry on cancer mortality risk estimates" by Preston et al. (Radiat. Res. 162, 377-389, 2004). PMID- 15799708 TI - An unbiased cell morphology-based screen for new, biologically active small molecules. AB - We have implemented an unbiased cell morphology-based screen to identify small molecule modulators of cellular processes using the Cytometrix (TM) automated imaging and analysis system. This assay format provides unbiased analysis of morphological effects induced by small molecules by capturing phenotypic readouts of most known classes of pharmacological agents and has the potential to read out pathways for which little is known. Four human-cancer cell lines and one noncancerous primary cell type were treated with 107 small molecules comprising four different protein kinase-inhibitor scaffolds. Cellular phenotypes induced by each compound were quantified by multivariate statistical analysis of the morphology, staining intensity, and spatial attributes of the cellular nuclei, microtubules, and Golgi compartments. Principal component analysis was used to identify inhibitors of cellular components not targeted by known protein kinase inhibitors. Here we focus on a hydroxyl-substituted analog (hydroxy-PP) of the known Src-family kinase inhibitor PP2 because it induced cell-specific morphological features distinct from all known kinase inhibitors in the collection. We used affinity purification to identify a target of hydroxy-PP, carbonyl reductase 1 (CBR1), a short-chain dehydrogenase-reductase. We solved the X-ray crystal structure of the CBR1/hydroxy-PP complex to 1.24 A resolution. Structure-based design of more potent and selective CBR1 inhibitors provided probes for analyzing the biological function of CBR1 in A549 cells. These studies revealed a previously unknown function for CBR1 in serum-withdrawal-induced apoptosis. Further studies indicate CBR1 inhibitors may enhance the effectiveness of anticancer anthracyclines. Morphology-based screening of diverse cancer cell types has provided a method for discovering potent new small-molecule probes for cell biological studies and anticancer drug candidates. PMID- 15799709 TI - Evolutionary origins of genomic repertoires in bacteria. AB - Explaining the diversity of gene repertoires has been a major problem in modern evolutionary biology. In eukaryotes, this diversity is believed to result mainly from gene duplication and loss, but in prokaryotes, lateral gene transfer (LGT) can also contribute substantially to genome contents. To determine the histories of gene inventories, we conducted an exhaustive analysis of gene phylogenies for all gene families in a widely sampled group, the gamma-Proteobacteria. We show that, although these bacterial genomes display striking differences in gene repertoires, most gene families having representatives in several species have congruent histories. Other than the few vast multigene families, gene duplication has contributed relatively little to the contents of these genomes; instead, LGT, over time, provides most of the diversity in genomic repertoires. Most such acquired genes are lost, but the majority of those that persist in genomes are transmitted strictly vertically. Although our analyses are limited to the gamma Proteobacteria, these results resolve a long-standing paradox-i.e., the ability to make robust phylogenetic inferences in light of substantial LGT. PMID- 15799710 TI - Systematic association of genes to phenotypes by genome and literature mining. AB - One of the major challenges of functional genomics is to unravel the connection between genotype and phenotype. So far no global analysis has attempted to explore those connections in the light of the large phenotypic variability seen in nature. Here, we use an unsupervised, systematic approach for associating genes and phenotypic characteristics that combines literature mining with comparative genome analysis. We first mine the MEDLINE literature database for terms that reflect phenotypic similarities of species. Subsequently we predict the likely genomic determinants: genes specifically present in the respective genomes. In a global analysis involving 92 prokaryotic genomes we retrieve 323 clusters containing a total of 2,700 significant gene-phenotype associations. Some clusters contain mostly known relationships, such as genes involved in motility or plant degradation, often with additional hypothetical proteins associated with those phenotypes. Other clusters comprise unexpected associations; for example, a group of terms related to food and spoilage is linked to genes predicted to be involved in bacterial food poisoning. Among the clusters, we observe an enrichment of pathogenicity-related associations, suggesting that the approach reveals many novel genes likely to play a role in infectious diseases. PMID- 15799713 TI - The reconstituted P-glycoprotein multidrug transporter is a flippase for glucosylceramide and other simple glycosphingolipids. AB - The Pgp (P-glycoprotein) multidrug transporter, which is linked to multidrug resistance in human cancers, functions as an efflux pump for non-polar drugs, powered by the hydrolysis of ATP at its nucleotide binding domains. The drug binding sites of Pgp appear to be located within the cytoplasmic leaflet of the membrane bilayer, suggesting that Pgp may function as a 'flippase' for hydrophobic compounds. Pgp has been shown to translocate fluorescent phospholipids, and it has been suggested that it may also interact with GlcCer (glucosylceramide). Here we use a dithionite fluorescence quenching technique to show that reconstituted Pgp can flip several NBD (nitrobenzo-2-oxa-1,3-diazole) labelled simple glycosphingolipids, including NBD-GlcCer, from one leaflet of the bilayer to the other in an ATP-dependent, vanadate-sensitive fashion. The rate of NBD-GlcCer flipping was similar to that observed for NBD-labelled PC (phosphatidylcholine). NBD-GlcCer flipping was inhibited in a concentration dependent, saturable fashion by various Pgp substrates and modulators, and inhibition correlated well with the Kd for binding to the protein. The addition of a second sugar to the headgroup of the glycolipid to form NBD-lactosylceramide drastically reduced the rate of flipping compared with NBD-PC, probably because of the increased size and polarity contributed by the additional sugar residue. We conclude that Pgp functions as a broad-specificity outwardly-directed flippase for simple glycosphingolipids and membrane phospholipids. PMID- 15799711 TI - Regulatory variation at glypican-3 underlies a major growth QTL in mice. AB - The genetic basis of variation in complex traits remains poorly understood, and few genes underlying variation have been identified. Previous work identified a quantitative trait locus (QTL) responsible for much of the response to selection on growth in mice, effecting a change in body mass of approximately 20%. By fine mapping, we have resolved the location of this QTL to a 660-kb region containing only two genes of known function, Gpc3 and Gpc4, and two other putative genes of unknown function. There are no non-synonymous polymorphisms in any of these genes, indicating that the QTL affects gene regulation. Mice carrying the high growth QTL allele have approximately 15% lower Gpc3 mRNA expression in kidney and liver, whereas expression differences at Gpc4 are non-significant. Expression profiles of the two other genes within the region are inconsistent with a factor responsible for a general effect on growth. Polymorphisms in the 3' untranslated region of Gpc3 are strong candidates for the causal sequence variation. Gpc3 loss of-function mutations in humans and mice cause overgrowth and developmental abnormalities. However, no deleterious side-effects were detected in our mice, indicating that genes involved in Mendelian diseases also contribute to complex trait variation. Furthermore, these findings show that small changes in gene expression can have substantial phenotypic effects. PMID- 15799714 TI - C-reactive protein and statins: IL-8 as a molecular link? AB - CRP (C-reactive protein) has not only emerged as a useful biomarker for cardiovascular disease, but also as a mediator of atherosclerosis. CRP directly activates vascular endothelial cells, amplifying the inflammatory response underlying atherogenesis. The expression of IL (interleukin)-8 appears to serve as one of the downstream effects of CRP. Kibayashi and co-workers in this issue of Clinical Science confirm that CRP induces IL-8 production in human aortic endothelial cells in vitro, via the activation of MAPKs (mitogen-activated protein kinases), an effect that can be inhibited by pitavastatin. PMID- 15799716 TI - Optimal conditions for the papain digestion of polyclonal ovine IgG for the production of bio-therapeutic Fab fragments. AB - In the present paper, we describe a rapid method for the determination of optimum conditions for papain digestion of polyclonal ovine IgG (purified by Na(2)SO(4) precipitation) for the production of bio-therapeutic Fabs (antigen-binding fragments). To determine the optimum conditions for digestion, a factorial approach to the design of experiments was undertaken. The resulting experimental data were used to construct the mathematical models using Design Expert 6.06(R) (Stat-Ease, Minneapolis, MN, U.S.A.) to predict the optimum conditions for a robust IgG digestion step. Optimum conditions were evaluated experimentally, and the applicability of the conditions for large-scale manufacture of bio therapeutic Fab fragments was assessed. The results and methods described in the present paper suggest that, provided the time and temperature are maintained at the high settings evaluated (24 h, 40 degrees C), the modelled data predict IgG digestion close to 100% for all the papain concentrations used. Provided papain is used at >2.5% (w/w), either time and/or temperature may be reduced. The results and methods described in the present paper may also be applicable to the generation of therapeutic Fab fragments from other immunoglobulins, including monoclonal antibodies purified from mammalian cell culture. PMID- 15799715 TI - Electrostatic stabilization in a pre-organized polar active site: the catalytic role of Lys-80 in Candida tenuis xylose reductase (AKR2B5) probed by site directed mutagenesis and functional complementation studies. AB - Lys-80 of Candida tenuis xylose reductase (AKR2B5) is conserved throughout the aldo-keto reductase protein superfamily and may prime the nearby Tyr-51 for general acid catalysis to NAD(P)H-dependent carbonyl group reduction. We have examined the catalytic significance of side-chain substitutions in two AKR2B5 mutants, Lys-80-->Ala (K80A) and Asp-46-->Asn Lys-80-->Ala (D46N K80A), using steady-state kinetic analysis and restoration of activity with external amines. Binding of NAD+ (Kd = 24 microM) and NADP+ (Kd = 0.03 microM) was 10- and 40-fold tighter in K80A than the wild-type enzyme, whereas binding of NADH (Kd = 51 microM) and NADPH (Kd = 19 microM) was weakened 2- and 16-fold in this mutant respectively. D46N K80A bound NAD(P)H and NAD(P)+ uniformly approx. 5-fold less tightly than the wild-type enzyme. The second-order rate constant for non covalent restoration of NADH-dependent reductase activity (kmax/Kamine) by protonated ethylamine was 0.11 M(-1).s(-1) for K80A, whereas no detectable rescue occurred for D46N K80A. After correction for effects of side-chain hydrophobicity, we obtained a linear free energy relationship of log (kmax/Kamine) and amine group pKa (slope = +0.29; r2 = 0.93) at pH 7.0. pH profiles of log (kcat/Km) for carbonyl group reduction by wild-type and D46N K80A revealed identical and kinetically unperturbed pKa values of 8.50 (+/-0.20). Therefore the protonated side chain of Lys-80 is not an essential activator of general acid catalysis by AKR2B5. Stabilized structurally through the salt-link interaction with the negatively charged Asp-46, it is proposed to pull the side chain of Tyr-51 into the catalytic position, leading to a preorganized polar environment of overall neutral charge, in which approximation of uncharged reactive groups is favoured and thus hydride transfer from NAD(P)H is strongly preferred. Lys-80 affects further the directional preference of AKR2B5 for NAD(P)H-dependent reduction by increasing NAD(P)H compared with NAD(P)+-binding selectivity. PMID- 15799717 TI - Effect of coffee on endothelial function in healthy subjects: the role of caffeine. AB - Coffee is one of the most widely used pharmacologically active beverages. The present study was designed to evaluate the acute effect of coffee ingestion on endothelial function in healthy individuals, and the potential role of caffeine. We studied 17 healthy young adults (28.9+/-3.0 years old; nine men), who were regular non-heavy coffee drinkers. The endothelial performance was estimated by endothelium-dependent FMD (flow-mediated dilatation) of the brachial artery before and 30, 60, 90 and 120 min after ingestion of a cup of caffeinated coffee (80 mg of caffeine) or the corresponding decaffeinated beverage (< 2 mg of caffeine) in two separate sessions, following a randomized single-blind cross over design. There was no difference in baseline FMD values between the two sessions [7.78 compared with 7.07% after caffeinated and decaffeinated coffee respectively; P = NS (not significant)]. Caffeinated coffee led to a decline of FMD (7.78, 2.86, 2.12, 4.44 and 4.57% at baseline, 30, 60, 90 and 120 min respectively; P < 0.001). This adverse effect was focused at 30 (P = 0.004) and 60 min (P < 0.001). No significant effect on FMD was found with the decaffeinated coffee session (7.07, 6.24, 5.21, 7.41 and 5.20%; P = NS). The composite effect of the type of coffee consumed over time on FMD was significantly different (P = 0.021). In conclusion, coffee exerts an acute unfavourable effect on the endothelial function in healthy adults, lasting for at least 1 h after intake. This effect might be attributed to caffeine, given that decaffeinated coffee was not associated with any change in the endothelial performance. PMID- 15799718 TI - Resolving the energy paradox of chaperone/usher-mediated fibre assembly. AB - Periplasmic chaperone/usher machineries are used for assembly of filamentous adhesion organelles of Gram-negative pathogens in a process that has been suggested to be driven by folding energy. Structures of mutant chaperone-subunit complexes revealed a final folding transition (condensation of the subunit hydrophobic core) on the release of organelle subunit from the chaperone-subunit pre-assembly complex and incorporation into the final fibre structure. However, in view of the large interface between chaperone and subunit in the pre-assembly complex and the reported stability of this complex, it is difficult to understand how final folding could release sufficient energy to drive assembly. In the present paper, we show the X-ray structure for a native chaperone-fibre complex that, together with thermodynamic data, shows that the final folding step is indeed an essential component of the assembly process. We show that completion of the hydrophobic core and incorporation into the fibre results in an exceptionally stable module, whereas the chaperone-subunit pre-assembly complex is greatly destabilized by the high-energy conformation of the bound subunit. This difference in stabilities creates a free energy potential that drives fibre formation. PMID- 15799719 TI - Structural characterization of the feline-immunodeficiency-virus envelope glycoprotein 36 ectodomain for the development of new antivirals. AB - In the fight against the human HIV, new targets are being explored, such as the proteins involved in the process of fusion of the virus with the host cell. Recently, the first generation of fusion inhibitors (enfuvirtide), targeting gp41 (virus envelope glycoprotein 41), has become commercially available. However, this promising class of drugs has to be improved in respect of their efficacy and bioavailability. Considering the strong homologies between HIV and FIV (feline immunodeficiency virus), as well as the highly conserved structure of the transmembrane envelope protein among species, FIV represents a relevant model of pre-screening studies for HIV. Taking into account (i) sequence homologies between the ectodomain of HIV gp41 and FIV gp36 (envelope glycoprotein 36), (ii) structural data available for gp41 and (iii) the fact that synthetic peptides derived from gp36 are effective inhibitors of FIV infection, we designed several peptides derived from gp36 sequence. We checked that these peptides had the same structural features as the corresponding peptides from gp41 HIV by CD, analytical ultracentrifugation and 1H-2H (hydrogen-deuterium) exchange combined with MS. By combining this latter technique with surface-plasmon-resonance assays, we identified the amino acid residues of the C-terminal region of the ectodomain of gp36 that are critical for interaction with the N-terminal region. This gave clues for therapy and vaccines against FIV, thus providing helpful data for treatments against HIV. PMID- 15799723 TI - Natural course of occult choroidal neovascularization in age-related macular degeneration: development of classic lesions in fluorescein angiography. AB - PURPOSE: To analyse the frequency of conversion from occult with no classic choroidal neovascularization (CNV) and occult with minimally classic CNV into predominantly classic CNV secondary to age-related macular degeneration. METHODS: Retrospective evaluation of baseline and repeat angiograms of 54 eyes with a follow-up of 6-12 months. RESULTS: In the group with initially occult with no classic CNV, nine of 40 eyes (23%) progressed to a predominantly classic lesion, whereas in the group of occult with a minimally classic CNV, 10 of 14 eyes (71%) eyes developed a predominantly classic CNV. CONCLUSIONS: The proportion of eyes that develop predominantly classic CNV is much higher in eyes with initially occult and minimally classic CNV, compared with eyes with initially occult and no classic CNV. Our data suggest that the natural course of CNV secondary to AMD begins as occult and progresses to classic CNV. PMID- 15799721 TI - Structure-activity studies with high-affinity inhibitors of pyroglutamyl peptidase II. AB - Inhibitors of PPII (pyroglutamyl-peptidase II) (EC 3.4.19.6) have potential applications as investigative and therapeutic agents. The rational design of inhibitors is hindered, however, by the lack of an experimental structure for PPII. Previous studies have demonstrated that replacement of histidine in TRH (thyrotropin-releasing hormone) with asparagine produces a competitive PPII inhibitor (Ki 17.5 microM). To gain further insight into which functional groups are significant for inhibitory activity, we investigated the effects on inhibition of structural modifications to Glp-Asn-ProNH2 (pyroglutamyl asparaginyl-prolineamide). Synthesis and kinetic analysis of a diverse series of carboxamide and C-terminally extended Glp-Asn-ProNH2 analogues were undertaken. Extensive quantitative structure-activity relationships were generated, which indicated that key functionalities in the basic molecular structure of the inhibitors combine in a unique way to cause PPII inhibition. Data from kinetic and molecular modelling studies suggest that hydrogen bonding between the asparagine side chain and PPII may provide a basis for the inhibitory properties of the asparagine-containing peptides. Prolineamide appeared to be important for interaction with the S2' subsite, but some modifications were tolerated. Extension of Glp-Asn-ProNH2 with hydrophobic amino acids at the C-terminus led to a novel set of PPII inhibitors active in vitro at nanomolar concentrations. Such inhibitors were shown to enhance recovery of TRH released from rat brain slices. Glp-Asn-Pro-Tyr-Trp-Trp-7-amido-4-methylcoumarin displayed a Ki of 1 nM, making it the most potent competitive PPII inhibitor described to date. PPII inhibitors with this level of potency should find application in exploring the biological functions of TRH and PPII, and potentially provide a basis for development of novel therapeutics. PMID- 15799724 TI - Transpupillary thermotherapy for occult subfoveal choroidal neovascularization: a 1-year, prospective randomized pilot study. AB - PURPOSE: To evaluate the 1-year effect of transpupillary thermotherapy (TTT) on occult choroidal neovascularization membranes (CNV). METHODS: In this prospective, randomized, controlled pilot study of 28 patients with occult or minimally classic (< 10%) neovascularization membranes with a diameter less than 4500 microm, 19 patients were treated with TTT, while nine received sham treatment. Outcome measures were membrane diameter, visual acuity and reading ability. RESULTS: The median age of patients randomized to TTT was 78 years (range 24 years); that of patients randomized to sham was 79 years (range 9 years). There was no difference regarding membrane diameter at baseline between the two groups; the median membrane diameters were 3400 microm (range 2400 microm) in the TTT group and 3200 microm (range 2300 microm) in the sham group (p = 0.639). Visual acuity (VA) was similar, with a median of 0.2 (minimum-maximum 0.08-0.5) in the TTT group and a median of 0.16 (min-max 0.10-0.32) in the sham group. A total of 21 patients were followed for 1 year, 13 in the TTT group (2.7 treatments/patient) and eight in the sham group. Membrane diameter increased in both groups, by a median of 350 microm (range 1600 microm) in the TTT group and 800 microm (range 1700 microm) in the sham group (p = 0.414), respectively, and there was a loss in VA of > or = 15 letters in 5/13 patients (38%) in the TTT group compared with 2/8 patients (25%) in the sham group (p = 0.266). Reading ability deteriorated equally over time in both groups. Seven patients were lost to follow-up due to reluctance to continue in the study (n = 4) or development of a classical component > 50% (n = 3) requiring photodynamic therapy (PDT). CONCLUSION: The results from this randomized, prospective pilot study of TTT for occult CNV did not indicate that TTT has a beneficial effect on visual outcome 1 year after treatment compared with the visual outcome that results from the natural course of the disease. The small study size limits statistical power and results from large control studies are needed. PMID- 15799725 TI - Topical dorzolamide increases pericentral visual function in age-related maculopathy: pilot study findings with short-wavelength automated perimetry. AB - PURPOSE: Numerous studies have confirmed the enhancement of ocular circulation by carbonic anhydrase inhibitors (CAIs). Topical CAI treatment with dorzolamide averts the significant pericentral visual function loss accompanying retinal and choroidal vasoconstriction during acute hyperventilation-induced hypocapnia. This study was designed to discern whether dorzolamide might similarly enhance macular function in patients with age-related maculopathy (ARM). METHODS: In a masked, placebo-controlled study, 40 patients with ARM and acuity > 20/50 were randomized to receive either dorzolamide or placebo for 12 weeks, thrice daily. After pre study perimetric training, pericentral function (mean sensitivity) was quantified using Humphrey 10-2 short-wavelength automated perimetry (SWAP), before and after 12 weeks of topical therapy. RESULTS: Dorzolamide-treated eyes demonstrated a significant increase in mean sensitivity of + 1.55 dB (p = 0.04); placebo-treated eyes showed no significant change (+ 0.58 dB; p = 0.10). Given the non significant increase of mean sensitivity in the placebo-treated group, fewer than 100 subjects per group would be required to afford > 70% power to yield a significant direct comparative difference between treatment and placebo in a prospective, randomized study of equally short duration. CONCLUSIONS: This study demonstrated a significant increase in short-wavelength sensitivity in ARM with dorzolamide and the lack thereof with placebo. These encouraging pilot study data suggest a potential role for topical CAIs in ARM patients, and establish objective parameters for prospective studies to further evaluate the effects of dorzolamide in ARM. PMID- 15799720 TI - Doxorubicin activates nuclear factor of activated T-lymphocytes and Fas ligand transcription: role of mitochondrial reactive oxygen species and calcium. AB - Doxorubicin (DOX), a widely used antitumour drug, causes dose-dependent cardiotoxicity. Cardiac mitochondria represent a critical target organelle of toxicity during DOX chemotherapy. Proposed mechanisms include generation of ROS (reactive oxygen species) and disturbances in mitochondrial calcium homoeostasis. In the present study, we probed the mechanistic link between mitochondrial ROS and calcium in the embryonic rat heart-derived H9c2 cell line and in adult rat cardiomyocytes. The results show that DOX stimulates calcium/calcineurin dependent activation of the transcription factor NFAT (nuclear factor of activated T-lymphocytes). Pre-treatment of cells with an intracellular calcium chelator abrogated DOX-induced nuclear NFAT translocation, Fas L (Fas ligand) expression and caspase activation, as did pre-treatment of cells with a mitochondria-targeted antioxidant, Mito-Q (a mitochondria-targeted antioxidant consisting of a mixture of mitoquinol and mitoquinone), or with adenoviral-over expressed antioxidant enzymes. Treatment with GPx-1 (glutathione peroxidase 1), MnSOD (manganese superoxide dismutase) or a peptide inhibitor of NFAT also inhibited DOX-induced nuclear NFAT translocation. Pre-treatment of cells with a Fas L neutralizing antibody abrogated DOX-induced caspase-8- and -3-like activities during the initial stages of apoptosis. We conclude that mitochondria derived ROS and calcium play a key role in stimulating DOX-induced 'intrinsic and extrinsic forms' of apoptosis in cardiac cells with Fas L expression via the NFAT signalling mechanism. Implications of ROS- and calcium-dependent NFAT signalling in DOX-induced apoptosis are discussed. PMID- 15799726 TI - Fundus autofluorescence in acute and chronic-recurrent central serous chorioretinopathy. AB - PURPOSE: Central serous chorioretinopathy (CSC) affects mostly young and middle aged adults. Typically, a serous retinal detachment occurs with a focal leakage point in fluorescein angiography. Fundus autofluorescence (AF) is related to the lipofuscin within the retinal pigment epithelium (RPE). As CSC is thought to be associated with RPE disorders, AF measurements might be able to detect distinct changes within the RPE level. The purpose of this study was to compare AF patterns in acute and chronic-recurrent CSC and to evaluate the potential value of AF as a non-invasive monitoring tool. METHODS: From 85 patients diagnosed with CSC (mean age 43 years), AF images were retrospectively evaluated and compared with angiographic and ophthalmoscopic findings. Fluorescein angiography and AF measurements were performed using a Heidelberg retina angiograph. Autofluorescence was excited at a wavelength of 488 nm and detected above 500 nm. RESULTS: Acute CSC as determined by an acute decrease in visual acuity (VA) within the last 6 weeks, focal point leakage and neurosensory retinal detachment was diagnosed in 39 patients. In 36 of those patients (92%), a significantly decreased AF at the leakage point (72%) and/or decreased AF in the area of neurosensory detachment (77%) were observed. Regarding chronic-recurrent CSC as determined by a decrease in VA for longer than 6 weeks and mottled hyperfluorescent appearance in angiography, abnormalities in AF were observed in 44/46 patients (96%). In those patients, decreased or mottled AF was observed at the leakage point itself (76%), whereas significantly increased AF was seen in the area of residual neurosensory retinal detachment (85%). CONCLUSIONS: In acute CSC, decreased AF is presumably due to a blockage caused by oedema, whereas in chronic-recurrent forms, irregular and increased AF is observed, possibly reflecting reactive RPE changes secondary to RPE defects and neurosensory detachment. If these findings could be systematically underlined by prospective clinical studies, AF might be an interesting non-invasive tool for monitoring RPE changes in CSC and for performing differential diagnosis. PMID- 15799727 TI - Metamorphopsia after successful retinal detachment surgery: an optical coherence tomography study. AB - PURPOSE: To investigate the findings in optical coherence tomography (OCT) in eyes with metamorphopsia after scleral buckling surgery for macula-off retinal detachment. METHODS: A total of 46 patients (46 eyes) with macula-off retinal detachment were prospectively studied 2 months after successful scleral buckling surgery. Patients were examined with the Amsler grid, fundus biomicroscopy and OCT. RESULTS: At the 2 month follow-up, 31/46 (67%) patients had metamorphopsia; 24/31 (77%) of these patients had an abnormal macular structure on OCT and 7/31 (23%) showed a normal macula. However, of the 15 patients in the non metamorphopsia group, 4/15 (27%) had an abnormal macula and 11/15 (73%) had a normal macula (p = 0.0015). The most common finding on OCT was neurosensory retinal detachment. CONCLUSIONS: Metamorphopsia after scleral buckling surgery in macula-off detachment is more common in patients in whom abnormal macular structure is seen on the OCT scan compared to those showing a normal macular structure. Neurosensory retinal detachment is the most common pathology in these patients. PMID- 15799728 TI - Combined phacoemulsification and pars plana vitrectomy for macular hole treatment. AB - PURPOSE: To assess the outcome of simultaneous phacoemulsification, pars plana vitrectomy and intraocular lens (IOL) implantation in eyes with macular hole. METHODS: A retrospective study was conducted in 38 eyes (36 patients) after combined phacoemulsification, insertion of a posterior capsule IOL and pars plana vitrectomy. RESULTS: The macular hole was successfully closed in 32 of the 38 eyes (84%). In six eyes (16%) the hole failed to close and one eye underwent a second operation. Vision improved by two or more Snellen lines in 29 eyes (73%), there was no change in seven eyes (18%), and visual acuity decreased in two eyes (5%). Intraoperative and postoperative complications included retinal tears in nine eyes (24%), posterior capsule rupture in two eyes (5%), transient postoperative increase of intraocular pressure in eight eyes (21%), and posterior capsule opacification in five eyes (13%). CONCLUSION: Combining phacoemulsification, IOL insertion and pars plana vitrectomy for macular hole repair can reduce the need for cataract surgery in the future, decrease costs, shorten postoperative recovery time and allow for clearer intraoperative visualization, making the procedure safer and more effective. PMID- 15799729 TI - Clinical outcomes of phacoemulsification cataract surgery in diabetes patients: visual function (VF-14), visual acuity and patient satisfaction. AB - PURPOSE: To evaluate the visual outcomes (visual acuity [VA] and visual function) after phacoemulsification cataract surgery in patients with diabetic retinopathy (DR), and assess patient satisfaction with final surgical outcome. METHODS: This prospective study comprised 74 eyes of 74 patients with different stages of DR. One surgeon (AW) performed all cataract surgery in a standardized fashion. Patients were assessed using the VF-14 (Visual Function-14) questionnaire. The following groups of patients were compared: those with no apparent retinopathy; those with mild non-proliferative DR (NPDR); those with severe NPDR, and those with proliferative DR (PDR). Visual acuity and visual function questionnaire (VF 14) responses were recorded preoperatively and 3 months postoperatively, during which the non-operated fellow eye showed no progression in retinopathy. RESULTS: Improvements in visual outcomes were significantly higher in groups 1 and 2 compared to groups 3 and 4 (Tukey-Kramer, p < 0.001). Comparisons between groups 1, 2 and 3, 4 showed significant differences in improvements in VA (Tukey-Kramer, p < 0.01), yet no statistically significant differences in functional (VF-14) improvements emerged between these groups. CONCLUSIONS: Patients with more advanced levels of DR showed no functional improvements despite improvements in VA. This emphasizes the relevance of patient education prior to surgery. In particular, it should be explained to patients with more advanced DR that, although surgery may be required, their functional improvement may be limited. PMID- 15799730 TI - Pharmacokinetics of intravitreal 5-fluorouracil prodrugs in silicone oil: experimental studies in pigs. AB - PURPOSE: To examine the in vivo pharmacokinetics of intravitreal 5-Fluorouracil (5-FU) following tamponade with 5-FU prodrug silicone oil formulations. METHOD: Two different alkoxycarbonyl 5-FU prodrugs denoted C12 and C18 were synthesized and formulated as silicone oil suspensions. A total of 26 pigs underwent conventional three-port lens-sparing pars plana vitrectomy. Approximately 1.6 ml of the prodrug-silicone oil formulation was placed in the vitreous cavity. Operated eyes were enucleated between 20 min and 168 hours postoperatively, and analysed for their content of free 5-FU by high performance liquid chromatography. RESULTS: With the C12 prodrug silicone oil formulation, the concentration of free 5-FU in the vitreous water phase 1 hour after surgery was 3.30 +/- 1.62 microg/ml. After 4 hours this concentration had declined to 1 microg/ml. With the C18 prodrug, the concentration of free vitreal 5-FU never reached 1 microg/ml during the 7 days these experiments lasted. A mathematical model is presented that can explain the measured data if the clearance of 5-FU from the vitreous water phase follows first order kinetics with a half-life of 20 min. CONCLUSION: These experiments, and the model analysis, suggest that the elimination half-life of 5-FU in the vitreous cavity of a vitrectomized, silicone oil-filled eye is very fast. The model analysis indicates that an alkoxycarbonyl 5-FU prodrug with a specific release rate constant of 10.7 microg/square root h cm(2) can maintain an intravitreal 5-FU concentration above 1 microg/ml for 5 days in the porcine eye. PMID- 15799731 TI - Central corneal thickness and vascular risk factors in normal tension glaucoma. AB - BACKGROUND: Normal tension glaucoma (NTG) has been shown to be associated with reduced central corneal thickness (CCT). The association of NTG with vascular risk factors is well documented. It has been postulated that a subset of NTG patients are misclassified due to incorrect intraocular pressure measurements on thin corneas. The aim of this study was to establish whether corneal thickness in NTG differs between patients with vascular risk factors specific to NTG and those without. METHODS: The study comprised a retrospective analysis of 108 eyes of 54 patients with NTG and 54 patients with primary open-angle glaucoma (POAG). Corneal thickness was measured in all patients. Vascular risk factors were recorded. Patients with NTG were divided into two groups depending on the presence (group A) or absence (group B) of vascular risk factors. RESULTS: The mean CCT was 549 +/- 34 microm in patients with POAG and 528 +/- 31 microm in patients with NTG (p = 0.001). Mean CCT was 512 +/- 31 microm in group A (n = 13) and 533 +/- 31 microm in group B (n = 41) (p = 0.034). A total of 40.9% of those with thin corneas (n = 22) had vascular risk factors versus only 12.5% of those with CCT within the normal range (n = 32) (p < 0.05). CONCLUSION: Central corneal thickness in NTG was significantly lower than in POAG and corneas were thinner in NTG patients with vascular risk factors than in those without. Vascular risk factors were significantly more common in patients with thin corneas. The finding of reduced corneal thickness in NTG does not obviate the need to consider vascular risk factors in the pathophysiology of the disease. PMID- 15799732 TI - Protocol for mitomycin C use in glaucoma surgery. AB - PURPOSE: To evaluate the results of a protocol described for mitomycin C (MMC) use in trabeculectomy or combined surgery (phacoemulsification and trabeculectomy). METHODS: A total of 143 eyes (60 trabeculectomies and 83 combined surgeries) of 124 patients were divided into four groups: group 1 (without MMC); group 2 (with 0.1 mg/ml MMC); group 3 (with 0.2 mg/ml MMC), and group 4 (with 0.4 mg/ml MMC). Two-minute MMC was used in every case in groups 2, 3 and 4. The results were analysed after 1 year of follow-up. Intraocular pressure (IOP) and complications were evaluated. Successful IOP control was defined when IOP was <21 mmHg and <16 mmHg if advanced glaucoma was present, always without additional medical treatment. RESULTS: Mean preoperative IOP decreased from 24.60 mmHg (SD 1.40 mmHg) to 13.47 mmHg (SD 0.37 mmHg) (p < 0.00001), 12 months postoperatively. Control in IOP was achieved in 79.02% of eyes. No significant differences were found in final mean IOP values (p > 0.196) or in postoperative complications (p > 0.120) in groups 2, 3 and 4. CONCLUSION: With the protocol described, a selection of concentration of MMC has been made in different clinical forms of glaucoma. No significant differences in IOP control and postoperative complications were noticed among the groups. PMID- 15799734 TI - Efficacy of lidocaine 2% gel in pterygium surgery. AB - PURPOSE: Anaesthesia during pterygium surgery is usually achieved with local injection of anaesthetic medications. The aim of this study was to determine whether lidocaine 2% gel is an effective topical anaesthetic agent for pterygium surgery. METHODS: We conducted a prospective, randomized clinical trial. Fifty four consecutive patients who had primary pterygium underwent surgical excision of the lesion and conjunctival autograft. Patients were randomized into two groups. Group 1 received 1 ml of lidocaine 2% solution subconjunctivally and group 2 received lidocaine 2% gel topically. The primary outcome of interest was the pain experienced during anaesthetic administration and surgery. Following the dissection and excision of the pterygium, conjunctival autograft was transplanted in each of the patients. Immediately after the operation, pain and discomfort scores were determined using a 10-point linear visual analogue scale. RESULTS: The mean pain score during anaesthetic administration was 4.26 +/- 1.18 in the injection group and 0.92 +/- 0.56 in the gel group. The mean pain score during surgery was 3.96 +/- 0.95 in the injection group and 4.0 +/- 1.01 in the gel group. There was a statistically significant difference in mean pain scores experienced during anaesthetic administration (p = 0.01). There was no significant difference in mean pain scores experienced during surgery (p = 0.55). No adverse events were noted. CONCLUSION: In light of these results, we conclude that topical administration of lidocaine 2% gel can be used as an alternative method of anaesthesia in pterygium surgery, especially for patients with needle phobia. PMID- 15799733 TI - Comparison of lidocaine 2% gel versus retrobulbar anaesthesia for implantation of Ahmed glaucoma drainage. AB - PURPOSE: To compare patient comfort and the efficacy of lidocaine 2% gel versus retrobulbar anaesthesia for Ahmed glaucoma implant surgery. PATIENTS AND METHODS: This prospective study included 32 eyes of 32 patients scheduled to undergo implantation of an Ahmed glaucoma drainage device for refractory glaucoma in one institution, randomized to receive either topical lidocaine 2% gel or retrobulbar anaesthesia. Sixteen eyes were randomized to each group. Patient assessment of pain was recorded during delivery of anaesthesia, during surgery and after surgery, using a visual analogue pain scale. The need for additional anaesthesia during tissue manipulation was recorded. Surgeon assessments of operative conditions, patient co-operation and intraoperative complications were also recorded. RESULTS: The patients in the retrobulbar group reported significantly more discomfort during administration of the anaesthetic agent than the patients in the topical anaesthesia group (p = 0.000). There was no significant difference in patient-reported pain scores intraoperatively (p = 0.317) or postoperatively (p = 0.875). Surgeon assessment showed similar levels of patient co-operation in both groups (p = 0.615). The overall operating conditions were overwhelmingly positive in each group (p = 0.186). The mean duration of surgery was significantly longer in the topical group compared with the retrobulbar group (p = 0.049). CONCLUSIONS: Lidocaine 2% gel is a safe alternative to retrobulbar anaesthesia for Ahmed drainage implant surgery and avoids the significantly greater pain associated with the administration of retrobulbar anaesthesia. It provides a reasonably safe and comfortable surgical environment and is well tolerated by patients. PMID- 15799735 TI - Molecular analysis of the NDP gene in two families with Norrie disease. AB - PURPOSE: To describe the molecular defects in the Norrie disease protein (NDP) gene in two families with Norrie disease (ND). METHODS: We analysed two families with ND at molecular level through polymerase chain reaction, DNA sequence analysis and GeneScan. RESULTS: Two molecular defects found in the NDP gene were: a missense mutation (265C > G) within codon 97 that resulted in the interchange of arginine by proline, and a partial deletion in the untranslated 3' region of exon 3 of the NDP gene. Clinical findings were more severe in the family that presented the partial deletion. We also diagnosed the carrier status of one daughter through GeneScan; this method proved to be a useful tool for establishing female carriers of ND. CONCLUSION: Here we report two novel mutations in the NDP gene in Mexican patients and propose that GeneScan is a viable mean of establishing ND carrier status. PMID- 15799736 TI - Basal cell carcinoma of the eyelid in Finland during 1953-97. AB - PURPOSE: To study the incidence of basal cell carcinoma (BCC) of the eyelid in Finland. METHODS: We studied 6241 cases of BCC of the eyelid reported to the nationwide Finnish Cancer Registry during 1953-97. We determined the age- and sex specific incidence rates and overall rates adjusted for age to the world standard population, and social class- and occupation-specific standardized incidence ratios, with the total Finnish population as reference. RESULTS: The incidence rates of BCC of the eyelid varied between 0.7 and 3.0 per 100 000 person-years in men and between 0.5 and 2.8 per 100 000 person-years in women during the study period. The age-adjusted incidence rates of BCC of the eyelid increased during 1953-87 (p < 0.0001). The incidence of BCC of the eyelid rose significantly with age. There were no significant differences in standardized incidence ratios (SIRs) for BCC of the eyelid between different social class and occupation categories. CONCLUSION: Age-adjusted incidence rates showed that BCC of the eyelid was more than twice as frequent during 1978-97 than before 1968. Ageing may partly explain the increased incidence of BCC of the eyelid, whereas there were no differences in the SIRs for BCC of the eyelid between different social class and occupation categories in Finland. PMID- 15799737 TI - Impact of UVR-A on whole human lenses, supernatants of buffered human lens homogenates, and purified argpyrimidine and 3-OH-kynurenine. AB - PURPOSE: Yellow chromophores and fluorescent compounds accumulate in the lens with age. Some of these compounds are photochemically active. The present study aimed to examine the photochemical effect of ultraviolet radiation-A (UVR-A) on the human lens. METHODS: Intact human lenses and supernatants of buffered lens homogenates were exposed to UVR-A. The effect of UVR-A was evaluated by time resolved and steady-state fluorescence spectroscopy, visual evaluation of colour and protein gel electrophoresis. RESULTS: Intact lenses exposed to UVR-A showed no changes in time-resolved or steady-state fluorescence properties but the yellow coloration was visibly attenuated. The supernatants of buffered lens homogenates exposed to UVR-A demonstrated a reduction in time-resolved and steady state fluorescent properties and protein cross-linking. CONCLUSIONS: Exposure of the intact lens to UVR-A causes chromophore bleaching without affecting fluorescence, indicating that non-fluorescent chromophores have been destroyed. After homogenization, both chromophores and fluorophores from the lens suffer damage and proteins aggregate. This indicates that powerful mechanisms of protection against UVR-A found in the intact lens are disturbed by homogenization of the lens, suggesting that isolated lens proteins cannot be used as a model system for studying cataractogenesis. Hypothetically, the protective mechanism could be related to the rigidly packed three-dimensional structure of the lens proteins or to the abundance of antioxidative and free radical scavenging defence systems. PMID- 15799738 TI - Ascorbate in the guinea pig lens: dependence on drinking water supplementation. AB - PURPOSE: To investigate whether lens ascorbate concentration can be elevated with drinking water supplementation. METHODS: Pigmented guinea pigs received drinking water supplemented with L-ascorbate, concentration 0.00, 2.84, 5.68 or 8.52 mm for a duration of 4 weeks. In addition, the chow fed to all animals contained 125 mmol L-ascorbate per kg of chow. At the end of the supplementation period, the guinea pigs were killed. Each lens was extracted. The lens was processed and ascorbate concentration was measured using high performance liquid chromatography (HPLC) with 254 nm ultraviolet radiation detection. The data were analysed with regression. RESULTS: At the end of the test period, all lenses were devoid of cataract as observed by slit-lamp examination. All lenses contained a detectable concentration of ascorbate. Estimated 95% confidence intervals for mean animal averaged lens ascorbate concentrations (micromol/g wet weight of whole lens) per group were 0.51 +/- 0.04 (0.00 mm; n = 6), 0.70 +/- 0.18 (2.84 mm; n = 6), 0.71 +/- 0.11 (5.68 mm; n = 5), and 0.71 +/- 0.06 (8.52 mm; n = 6). Animal-averaged lens ascorbate concentration [Asc(lens)] (micromol/g wet weight lens) increased with ascorbate supplementation in drinking water [Asc(water)] (M), in agreement with the model: [Asc(lens)] = A - Be(-kAsc(water)]. CONCLUSION: Lens ascorbate concentration increases with drinking water supplementation in the guinea pig without cataract development. The currently presented method for measurement of whole lens ascorbate content is suitable. PMID- 15799740 TI - Surgical drainage of submacular haemorrhage from ruptured retinal arterial macroaneurysm. PMID- 15799741 TI - YAG curios #1: repeat Nd:YAG laser posterior capsulotomy. AB - A descriptive case series of five patients identified either opportunistically or from a retrospective review of the laser register is presented. All underwent cataract surgery between 1995 and 1999 and required initial Nd:YAG laser posterior capsulotomy 15-30 months after surgery. They subsequently developed reclosure of the capsulotomy, necessitating repeat laser capsulotomy 11-82 months later. One patient, who received a hydrogel intraocular lens, required a third capsulotomy after a further 12 months. The rate of repeat Nd:YAG laser capsulotomy in our unit was 0.31%. PMID- 15799739 TI - Soluble CD163 and interleukin-6 are increased in aqueous humour from patients with endothelial rejection of corneal grafts. AB - PURPOSE: To evaluate soluble CD163 (sCD163) as a new marker of macrophage activity in aqueous humour from patients with corneal rejection and to investigate correlations between sCD163, the CD163 inducing interleukin-6 (IL-6), and albumin; to investigate whether increases in sCD163 and IL-6 levels in aqueous humour were results of intra- or extraocular production, and to investigate the impact of sCD163 and IL-6 levels on graft outcome. METHODS: Aqueous humour was obtained from 19 patients with endothelial rejection of corneal grafts, seven cataract patients and five cataract patients with uncomplicated corneal grafts. The presence of sCD163 was investigated by immunoprecipitation and immunoblotting. The concentrations of sCD163, IL-6 and albumin were measured by ELISA. RESULTS: Soluble CD163 was detected in aqueous humour from patients with corneal rejection. Soluble CD163, IL-6 and albumin were significantly increased in aqueous humour from patients with corneal rejection when compared with control patients. When normalizing sCD163 and IL-6 with albumin, the differences between rejection patients and control groups became insignificant for sCD163 but remained significant for IL-6. Neither IL-6 nor sCD163 were related to the outcome of the corneal rejection. CONCLUSIONS: Both sCD163 and IL-6 are present in high levels in aqueous humour from patients with rejection of corneal grafts. Our data suggest that IL-6 is produced locally, whereas sCD163 is at least partly derived from influx from plasma. PMID- 15799742 TI - YAG curios #2: Nd:YAG laser treatment following suction posterior capsulorhexis. AB - Suction posterior capsulorhexis (SPC) is a relatively new technique for creating a posterior capsulorhexis. We present a case series of seven patients who required Nd:YAG laser treatment 27 months after SPC. No patient had pre-existing risk factors for increased postoperative inflammation and all received a one piece polymethylmethacrylate intraocular lens placed in-the-bag. Although it reduces the need for Nd:YAG laser treatment, SPC, like the more usual forceps posterior capsulorhexis, does not completely eliminate it. PMID- 15799743 TI - Intravitreal triamcinolone acetonide for treatment of cystoid macular oedema in patients with retinitis pigmentosa. AB - PURPOSE: To evaluate the anatomic and visual outcomes of intravitreal triamcinolone acetonide injection in patients with cystoid macular oedema (CMO) secondary to retinitis pigmentosa (RP). METHODS: Five eyes of five patients with CMO secondary to RP, aged 25-41 years (mean 33.2 years) made up the study population. All eyes had persistent CMO despite medical treatment with 250 mg of oral acetazolamide twice daily for 1 month. Intravitreal injection of 4 mg (0.1 ml) triamcinolone acetonide was offered to treat macular oedema. The visual and anatomic responses were observed, as well as complications related to the injection procedure and corticosteroid medication. RESULTS: Follow-up periods varied between 6 and 8 months (mean 6.8 months); all patients completed 6 months of follow-up. After intravitreal triamcinolone acetonide injection all patients showed good anatomic response. The baseline median central macular thickness was 418 microm (range 376-626 microm). At 1 month, the median central macular thickness had decreased to 224 microm (range 214-326 microm). At 3 and 6 months, the median central macular thicknesses were 275 microm (range 215-584 microm) and 312 microm (range 239-521 microm), respectively. Recurrent CMO was found in one patient at the 3-month follow-up and in two patients at the 6-month follow-up. Retreatment was performed in these patients. At the 1-month follow-up, no patient was found to have lost vision and two patients showed improvement. At the 3- and 6-month follow-ups, no patient had lost vision from baseline but no patient had maintained their improved visual acuity (VA). CONCLUSIONS: In our small series, all patients showed an anatomic improvement in CMO after intravitreal injection of triamcinolone acetonide. However, in three out of five patients, despite good anatomic results, no improvement in VA was achieved. Because of the limitations of this pilot study, it is difficult to explain why no improvement in VA was achieved despite good anatomic results in some patients. Further study with longer follow-up periods and larger series is warranted to assess the efficacy of the treatment. PMID- 15799744 TI - Capsular bag distension syndrome after combined cataract and glaucoma surgery. AB - PURPOSE: To report four cases of capsular bag distension syndrome (CBDS) after combined cataract and glaucoma surgery. METHODS: We describe the clinical features and evolution of each case after individual treatment. CASE REPORTS: We report four cases of CBDS after phacoemulsification, combined with trabeculectomy in two cases and with an Ahmed aqueous drainage device implantation in the other two cases. The space between the intraocular lens (IOL) and the posterior capsule was occupied by an optically clear liquid in two cases and by a turbid liquid in the other two cases, with posterior bowing of the capsule into the anterior vitreous. A myopic shift and anterior chamber shallowing occurred in three cases. After posterior Nd:YAG laser capsulotomy, the CBDS resolved in three cases. In case 1, a surgical posterior capsulotomy and anterior vitrectomy were necessary. DISCUSSION: Capsular bag distension syndrome should be included in the differential diagnosis of a shallow anterior chamber after combined cataract and glaucoma surgery. PMID- 15799745 TI - Valsalva retinopathy: diagnostic challenges in a patient with pars-planitis. AB - PURPOSE: To describe a case of Valsalva retinopathy in a patient with pars planitis and to discuss its management. METHODS: We present a case of a 19-year old male with a six-month history of bilateral intermediate uveitis, referred for evaluation of recent-onset spontaneous bilateral vitreous hemorrhages. Clinical evaluation and fluorescein angiography were negative for the presence of neovascularization. On closer questioning the patient revealed that he had his most recent symptoms after heavy weight lifting. RESULTS: The patient was instructed to avoid heavy weight lifting. Over the next two months, vitreal hemorrhages had resolved. However, despite the treatment with systemic prednisone, the patient's intermediate uveitis was still active, and cystoid macular edema had developed. Therefore, we proceeded to systemic immunomodulatory therapy, which controlled the inflammation and preserved vision in each eye. CONCLUSIONS: Although a very rare cause, Valsalva maneuver should be included in the differential diagnosis of bilateral vitreous hemorrhages. Ophthalmologists should advise patients with known eye problems that can predispose to vitreal hemorrhages to refrain from it. PMID- 15799747 TI - Explantation of an acrylic Acrysof intraocular lens without wound enlargement. PMID- 15799746 TI - Cytomegalovirus retinitis in Wegener's granulomatosis: case report and review of the literature. AB - PURPOSE: To present the first case of cytomegalovirus (CMV) retinitis in a Wegener's granulomatosis patient on systemic cyclosporine A (CsA) therapy. METHODS: A 41-year-old patient was treated with systemic CsA for c-ANCA positive Wegener's granulomatosis. Four weeks later he presented with blurred vision and a retinitis was diagnosed in the right eye. After detection of CMV DNA by polymerase chain reaction (PCR), the patient was treated with intravenous gancyclovir. Due to bone marrow toxicity, systemic antiviral therapy was discontinued and an intravitreal gancyclovir pellet was implanted. RESULTS: The retinitis lesions were scarified under this therapy and the patient's visual function was restored completely. CONCLUSIONS: Cytomegalovirus retinitis can occur under systemic monotherapy with CsA in patients with systemic vasculitis. Intraocular virus DNA detection by PCR is recommended to rule out retinal involvement of the underlying vasculitis in atypical cases and to preserve the patient's visual acuity. PMID- 15799749 TI - Bullous keratopathy treated with honey. PMID- 15799750 TI - Effects of perfluorohexyloctane in the anterior chamber. PMID- 15799751 TI - Neurosyphilitic episcleritis. PMID- 15799752 TI - Dural ectasia of the optic nerve sheath. PMID- 15799753 TI - Optical coherence tomography findings in macular cavernous haemangioma. PMID- 15799754 TI - Abducens nerve paresis in spontaneous intracranial hypotension syndrome. PMID- 15799757 TI - Bacterial vaginosis--a laboratory and clinical diagnostics enigma. AB - Diagnosing bacterial vaginosis (BV) has long been based on the clinical criteria of Amsel et al., whereby three of four defined criteria must be satisfied. Though there are other criteria and scoring methods which function well in comparison (i.e. Nugent scoring), it is not certain that they will always identify the same category of patients. Point-of-care methods based on various combinations of microbial products, presence of RNA, or more complex laboratory instrumentations such as sensor arrays, have also been introduced for the diagnosis of BV. No method for diagnosing BV can at present be regarded as the best. It could be that -based partly on tacit knowledge on the part of the clinical investigators scoring in the clinic--various scoring systems have been chosen to fit a particular BV-related problem in a particular population. In this review we critically examine these pertinent issues influencing clinical scoring and laboratory diagnostics of BV. PMID- 15799758 TI - The value of tartrate resistant acid phosphatase (TRAP) immunoreactivity in diagnosis of hairy cell leukemia. AB - Hairy cell leukemia (HCL) is a rare chronic B-cell lymphoproliferative disorder characterized by splenomegaly, pancytopenia, and circulating atypical lymphocytes with circumferential cytoplasmic projections. We investigated the specificity and the sensitivity of anti-TRAP antibody immunoreactivity in 57 cases of HCL. We found that there is a statistically highly significant difference between TRAP immunoreactivities of the study and the control groups, and HCL can be diagnosed by TRAP immunoreactivity in bone marrow trephine biopsy materials with a specificity of 98.27 % and a sensitivity of 100%. PMID- 15799759 TI - Complement activation in acute coronary syndromes. AB - The complement system is part of the host defence response. However, considerable evidence suggests that complement plays an important role in the pathophysiology of ischemic heart disease. The aim of this study was to evaluate complement activation in patients with all forms of acute coronary syndromes (ACS) and to examine the relationship between the degree of complement activation and myocardial injury. The study population included 152 subjects (26 females): 82 with ACS (35 acute myocardial infarction (AMI), 22 non-Q wave MI (NQMI), 25 unstable angina (UAP)) (Group A), 35 stable angina (SA) (Group B), and 35 healty control subjects (Group C). Complement 3 (C3), Complement 4 (C4), C-reactive protein (CRP), troponin I (TnI) as well as creatine kinase MB (CK-MB) were evaluated. Patients' blood samples were taken on admission (day 1) and after 2, 3 and 7 days in group A. However, only one measurement was performed in the groups B and C. Plasma C3 and C4 peak levels were significantly higher in patients with AMI (141+/-29 and 35+/-11 mg/dl) and NQMI (136+/-13 and 35+/-7 mg/dl) than in patients with SA (128+/-14 and 27+/-10 mg/dl) and the control subjects (114+/-22 and 22+/-7 mg/dl) (p<0.03). Also, C3 and C4 serum levels in patients with SA and UAP (126+/-16 and 31+/-7 mg/dl) were significantly higher than those in control subjects (p<0.01, p<0.03, respectively). At 1-week follow-up, there were no significant differences between the plasma levels of C3 and C4 in patients with UAP (p>0.05). However, plasma levels of C3 and C4 were significantly different between days in patients with AMI and NQMI (p<0.0001). Plasma C3 and C4 levels in ACS showed a relationship with peak CK-MB and Tn I levels (p<0.01). Plasma CRP level in ACS showed positive correlation with C3 (p<0.01) and C4 (p<0.001). In this study, we determined that plasma C3 and C4 levels were elevated in ACS and SA. Although C3 and C4 were higher in ACS and SA, the systemic levels of inflammatory markers in patients with SA and UAP were lower than those found in the AMI and NQMI groups. The relationship between C3, C4 levels and ACS further suggests that the complement activation is related to necrosis within the myocardium. PMID- 15799760 TI - Comparison of HER-2/neu, ER and PCNA expression in premenopausal and postmenopausal patients with breast carcinoma. AB - We attempted to compare the pattern of HER-2/neu, ER and PCNA in premenopausal and postmenopausal patients with breast carcinoma to identify potential biological differences. Five hundred and forty-eight samples from 318 premenopausal and 230 postmenopausal women with invasive ductal carcinoma of the breast were evaluated for HER-2/neu, ER and PCNA expression by immunohistochemistry. HER-2/neu expression showed 27.4% positivity in premenopausal and 24.8% in postmenopausal women; there was no significant difference between the two groups (p>0.05). In contrast, HER-2/neu expression was found to be significantly associated with ER negativity in the two groups (p<0.05 in premenopausal, p<0.001 in postmenopausal patients). However, it was significantly associated with PCNA expression only in the postmenopausal group (p<0.001). 54.4% showed premenopausal tumor cell ER positivity, whereas 64.3% of the postmenopausal group showed positivity. ER expression showed a significant correlation with patient menopausal status (p<0.05). The prevalence of PCNA positivity in the tumor cell components is slightly higher in postmenopausal compared to premenopausal women (p>0.20). The current study is consistent with reports from other groups regarding the correlation of HER-2/neu with adverse pathologic features and with expression of other markers in carcinoma. We also observed there was no trend toward increased HER-2/neu expression in either premenopausal or postmenopausal patients, i.e. there was similar HER-2/neu expression in the two groups. This suggests that HER-2/neu status could be used to determine assignment to specific intensive adjuvant therapy and evaluation of biological behavior in both pre- and postmenopausal patients with breast carcinoma. PMID- 15799761 TI - Pharyngeal carriage of serogroup W135 Neisseria meningitidis in Hajjees and their family contacts in Morocco, Oman and Sudan. AB - In 2000 the global outbreak that began in Saudi Arabia was caused by a W135:2a:P1.5,2 strain of Neisseria meningitidis belonging to the ET-37 complex and to ST-11. There was concern that introduction of this epidemic clone (EC) might lead to a wave of outbreaks in the African meningitis belt. The WHO therefore initiated studies of meningococcal carriage among pilgrims and their family contacts in Morocco, Oman and Sudan, 3 to 12 months after the Hajj 2000. In Morocco, 1186 persons were swabbed 3 times. Ninety-five meningococcal strains were isolated from 2.7% of the specimens. Pulsed-field gel electrophoresis showed that 32 (33.6%) were identical with the EC. In Sudan, 5 strains identical with the EC were obtained after sampling 285 persons. In Oman, among 18 meningococcal strains isolated from 399 subjects, 11 (61.1%) belonged to the EC. The important pharyngeal carriage of W135 (EC) and its role in the 2001-2002 outbreaks in Burkina Faso argues for the necessity of reinforcing surveillance, and adapting and planning responses in Africa and the Middle East using the most appropriate vaccine. PMID- 15799762 TI - Carbapenem resistance mechanisms in Pseudomonas aeruginosa: alterations of porin OprD and efflux proteins do not fully explain resistance patterns observed in clinical isolates. AB - Imipenem resistance in Pseudomonas aeruginosa is considered to be associated with loss of the porin OprD combined with activity of chromosomal beta-lactamase (AmpC), while overexpression of multidrug efflux pumps is considered to confer meropenem resistance. Carbapenem resistance can also result from production of metallo-beta-lactamases. Transcription of oprD and efflux pump genes mexB, mexY and mexF was analysed in 23 clinical isolates of P. aeruginosa by quantitative RT PCR. oprD was sequenced in all, and mexR, regulator of efflux pump MexAB-OprM, in selected isolates. Four isolates that were imipenem susceptible had significant reduction of oprD mRNA and presence of oprD mutations causing frameshift or translational stop. In strains only resistant to imipenem no significant difference in transcription of oprD was observed between low-level and high-level resistant isolates. The differences could not be explained by either pattern of oprD mutations. Increased transcription of mexB generally correlated well with meropenem resistance. One high-level meropenem-resistant isolate showed no significant change in mexB mRNA, but sequencing confirmed presence of a nalB mutation. Furthermore, one meropenem-susceptible isolate showed significant increase in mexB transcription, but no mexR mutations. In summary, our findings indicate that the resistance patterns observed cannot be fully explained by the currently described carbapenem resistance mechanisms. PMID- 15799763 TI - Cross-reactivity between human adenoviruses in delayed-type hypersensitivity. AB - The aim of this study was to determine the antigen responsible for the induction of delayed-type hypersensitivity (DTH) by human adenoviruses (Ads). The estimation of DTH was based on measurement of the extent of swelling of the hind footpads of mice. CsCl density gradient-purified human Ad serotype 6 (Ad6) induced DTH in a dose-dependent manner. In Ad6-sensitized mice, DTH could be elicited by serotypes belonging to the same species of human Ads (types 1 and 5) and by a serotype (type 3) belonging to another species. Latex particles coated with purified hexon antigen prepared from Ad5 had the capacity to sensitize mice and elicit a DTH reaction. We suggest that, for serotypes belonging to species C, the cross-reactive highly conserved T cell epitope of the hexon protein might be responsible for the DTH induction, and furthermore the same epitope might result in the cross-reactivity between serotypes 3 and 6. The possible importance of these data is discussed in relation to human gene therapy through the application of Ad vectors. PMID- 15799764 TI - Lack of agreement between biochemical and genetic identification of Aeromonas spp. AB - Biochemical and genetic identification by RFLP (restriction fragment length polymorphism) of the PCR-amplified 16S r-RNA sequence were compared for a selection of 171 clinical and environmental isolates of Aeromonas spp. The investigation revealed large differences between the two methods. The species phenotypic identification scheme and the genetic technique applied to the environmental strains gave divergent results for 96% of the strains tested. There was 46% discrepancy between the two methods for the clinical isolates. The distribution of species differed between clinical and environmental isolates. A. hydrophila, A. caviae, A. jandaei and A. veronii dominated the clinical material (81% of isolates by RFLP), whilst only 21% of the environmental isolates belonged to those four species. From the environmental group A. salmonicida, A. bestiarum, A. sobria, A. media, and A. encheleia contributed 72% of the strains tested. The poor parity between the biochemical and the genetic identification of the environmental isolates, and to a lesser extent for the clinical isolates, underlines the fact that our current biochemical methods cannot adequately differentiate Aeromonas spp. This work also shows that the biochemical schemes derived from clinical isolates are incomplete for the identification of environmental strains. PMID- 15799765 TI - The value of cytology in the diagnostics of lung cancer. AB - In order to elucidate the relative contributions made by cytology and histology in the diagnosis of lung cancer, we studied the cytology and histology reports of all patients who received a microscopic diagnosis of lung cancer in our hospital during the 7 years 1996-2002. This gave a total of 407 patients. The most frequent diagnoses were squamous cell carcinoma (34.9%), adenocarcinoma (24.8%), and small cell carcinoma (17.8%). One hundred and fifteen patients (28.3%) received their microscopic diagnosis based only on cytology, which therefore proved to be of great diagnostic value. The most useful type of cytology specimen was taken by bronchial lavage or bronchial brushing. These types of specimens provided the diagnosis in 71 patients (17.4%). Cytology was especially capable of finding squamous cell carcinomas. Small cell carcinomas were underrepresented (9.6% versus 17.8%) and unspecified carcinomas greatly overrepresented (9.6% versus 2.9%) among the diagnoses obtained by cytology alone. We conclude that cytology is of considerable diagnostic value, although not as specific as histology for the subtyping of carcinomas. Clinicians should be more aware of the usefulness of cytology, especially in cases where it is difficult to obtain bronchoscopic biopsy samples for histological examination. PMID- 15799766 TI - Weekly point prevalence of Streptococcus pneumoniae, Hemophilus influenzae and Moraxella catarrhalis in the upper airways of normal young children: effect of respiratory illness and season. AB - The aim was to determine the effect of respiratory illness and season on carriage rates in the upper airways of Streptococcus pneumoniae, Hemophilus influenzae and Moraxella catarrhalis in normal children. Sixteen healthy children, 1-10 years old, amenable to weekly sampling were followed longitudinally for at least three seasons of the year. Respiratory symptoms were recorded daily; weekly nasal aspirate/wash samples were cultured on selective agars. Urea concentration in samples was used to define dilution of secretion. 68% of 950 samples were culture positive; 44% of positives had two or all three species. Each species was detected in about one third of samples. Bacteria were detected in 76% of samples during illness vs. 65% during wellness (p=0.004). Seasonal carriage rates varied from 56% in summer and fall to 85% in winter. There was a strong inverse correlation between dilution of secretion and bacterial detection rate in illness and wellness aspirate samples during the four seasons (r=-0.82, p=0.01). Detection of bacteria varied with the amount of secretion in the sample. This variation accounts for the apparent differences in bacterial carriage during illness vs. wellness and during different seasons. PMID- 15799767 TI - Follicular lymphoid hyperplasia of the oral cavity representing progressive transformation of germinal center. AB - Follicular lymphoid hyperplasia (FLH) of the oral cavity is a rare and poorly understood lymphoproliferative disorder. We present a case of FLH of the oral cavity presenting with progressive transformation of germinal center (PTGC). The patient was a 49-year-old Japanese woman presenting with a hard mass in the right cheek. The resected specimen contained numerous lymphoid follicles with active germinal centers and a portion of the lymphoid follicles exhibited PTGC. The PTGCs contained a few large lymphoid cells resembling lymphocytic and histiocytic Reed-Sternberg cells of nodular lymphocyte-predominant Hodgkin lymphoma. The PTGC was surrounded by groups of epithelioid cells. In situ hybridization studies demonstrated strong expression of Epstein-Barr virus (EBV)-encoded small RNA in scattered large lymphoid cells in the PTGC. Although the etiology of FLH of the oral cavity remains unclear, the present case suggests that a subset of FLH of the oral cavity appears to be an EBV-associated lymphoproliferative disorder. PMID- 15799768 TI - Brain abscess due to Actinobacillus actinomycetemcomitans. AB - Actinobacillus actinomycetemcomitans, a constituent of the oral flora, is a rare cause of brain abscesses. We report the case of a 47-year-old male who presented with multiple brain abscesses due to this organism, presumably originating from his poor dentition. Problems met in isolating and identifying A. actinomycetemcomitans suggest that its true rate of isolation from non-oral samples may have been underestimated. PMID- 15799769 TI - Dysgonomonas capnocytophagoides bacteraemia in a neutropenic patient treated for acute myeloid leukaemia. AB - Dysgonomonas capnocytophagoides, formerly known as CDC group DF-3, is an opportunistic pathogen associated with diarrhoea and very rarely bacteraemia. We report a case of D. capnocytophagoides found in blood cultures from a severely neutropenic patient treated for acute myeloid leukaemia. The isolate was found resistant to penicillin, cephalosporins, meropenem, aminoglycosides and ciprofloxacin, and susceptible to ampicillin, tetracycline, chloramphenicol, clindamycin and trimethoprim-sulphamethoxazole. It was identified using conventional phenotypic testing but remained unidentified by the automated identification system (Vitek-2) as this system did not contain DF-3 or D. capnocytophagoides in its database. PMID- 15799770 TI - Non-invasive monitoring of muscle blood perfusion by photoplethysmography: evaluation of a new application. AB - AIM: To evaluate a specially developed photoplethysmographic (PPG) technique, using green and near-infrared light sources, for simultaneous non-invasive monitoring of skin and muscle perfusion. METHODS: Evaluation was based on assessments of changes in blood perfusion to various provocations, such as post exercise hyperaemia and hyperaemia following the application of liniment. The deep penetrating feature of PPG was investigated by measurement of optical radiation inside the muscle. Simultaneous measurements using ultrasound Doppler and the new PPG application were performed to elucidate differences between the two methods. Specific problems related to the influence of skin temperature on blood flow were highlightened, as well. RESULTS: Following static and dynamic contractions an immediate increase in muscle perfusion was shown, without increase in skin perfusion. Liniment application to the skin induced a rapid increase in skin perfusion, but not in muscle. Both similarities and differences in blood flow measured by Ultrasound Doppler and PPG were demonstrated. The radiant power measured inside the muscle, by use of an optical fibre, showed that the near-infrared light penetrates down to the vascular depth inside the muscle. CONCLUSIONS: The results of this study indicate the potentiality of the method for non-invasive measurement of local muscle perfusion, although some considerations still have to be accounted for, such as influence of temperature on blood perfusion. PMID- 15799771 TI - Short-term effects of repetitive arm work and dynamic exercise on glucose metabolism and insulin sensitivity. AB - AIM: To determine whether repetitive arm work, with a large component of static muscle contraction alters glucose metabolism and insulin sensitivity. METHOD: Euglycemic clamps (2 h) were started in ten healthy individuals 15 min after 37 min periods of: (1) repetitive arm work in a simulated occupational setting; (2) dynamic concentric exercise on a cycle ergometer at 60% of VO(2max) and (3) a resting regime as a control. During the experimental periods, blood samples were collected, blood pressure was measured repeatedly and electrocardiogram (ECG) was recorded continuously. During the clamps, euglycemia was maintained at 5 mmol l( 1) and insulin was infused at 56 mU m(-2) min(-1) for 120 min. RESULTS: The insulin-mediated glucose disposal rate (M-value) for the steady-state period (60 120 min) of the clamp, tended to be lower following arm work than for both cycling and resting regimes. When dividing the steady-state period into 20-min intervals, the insulin sensitivity index (ISI) was significantly lower for arm work compared with the resting control situation between 60-80 min (P = 0.04) and 80-100 min (P = 0.01), respectively. Catecholamines increased significantly for arm work and cycling compared with resting regime. Data from heart rate variability (HRV) measurements indicated significant sympathetic activation during repetitive arm work. CONCLUSION: The results indicate that repetitive arm work might acutely promote insulin resistance, whereas no such effect on insulin resistance was produced by dynamic concentric exercise. PMID- 15799773 TI - Transcriptional responses of rat skeletal muscle following hypoxia-reoxygenation and near ischaemia-reperfusion. AB - AIM: The effect of ischaemia/reperfusion or hypoxia/reoxygenation on gene expression has not been extensively studied. We hypothesized that in skeletal muscle, tissue hypoxia of similar magnitude but induced by different mechanisms would lead to different transcriptional responses. METHODS: Muscle gene transcription was assessed using microarray analysis and reverse transcriptase polymerase chain reaction in 18 rats exposed to regional hind limb near ischaemia/reperfusion (n = 6), hypoxia/reoxygenation (n = 6) or sham operation (n = 6). Hypoxic burden was measured by the area under the PtO(2)-time curve. RESULTS: PtO(2) was reduced in both the near ischaemia/reperfusion and hypoxia/reoxygenation groups. Although the hypoxic burden was similar, the genomic response was different for each condition. Near ischaemia/reperfusion had a greater effect on gene expression than hypoxia/reoxygenation. Using stringent criteria for changes in gene expression (i.e. more than or equal to twofold change vs. control) unique patterns of gene expression could be identified suggesting individualized transcriptional responses to each of these injuries. Several genes, including insulin-like growth factor 1 (IGF-1) and cyclin dependent kinase inhibitor (p27(Kip1)) were induced by both injury types and these may have potential clinical application as markers of tissue damage. In contrast, no single gene was downregulated by both injury conditions. CONCLUSIONS: The mechanism of skeletal muscle hypoxia has a profound effect on its subsequent transcriptional response. We identified several potential candidates as markers of skeletal muscle ischaemic damage. PMID- 15799772 TI - Chronic aerobic exercise enhances components of the classical and novel insulin signalling cascades in Sprague-Dawley rat skeletal muscle. AB - AIM: The aim of this study was to provide a more extensive evaluation of the effects of chronic aerobic exercise on various components of the insulin signalling cascade in normal rodent skeletal muscle because of the limited body of literature that exists in this area of investigation. METHODS: Male Sprague Dawley rats were assigned to either control (n = 7) or chronic aerobic exercise (n = 7) groups. Aerobic exercise animals were run 3 day week(1) for 45 min on a motor-driven treadmill (32 m min(1), 15% grade) for a 12 week period. Following the training period, all animals were subjected to hind limb perfusion in the presence of 500 microU mL(1) insulin to determine what effect chronic aerobic training had on various components of the insulin signalling cascade, c-Cbl protein concentration and c-Cbl phosphorylation. RESULTS: Twelve weeks of aerobic training did not alter skeletal muscle Akt 1/2 protein concentration, Akt Ser 473 phosphorylation, Akt Thr 308 phosphorylation, Akt 1 activity, aPKC-zeta protein concentration, aPKC-lambda protein concentration or c-Cbl protein concentration. In contrast, chronic aerobic exercise increased insulin-stimulated phosphatidylinositol 3-kinase, Akt 2 kinase and aPKC-zeta/lambda kinase activities, as well as c-Cbl tyrosine phosphorylation, in a fibre type specific response to aerobic training. In addition, chronic aerobic exercise enhanced insulin-stimulated plasma membrane glucose transporter 4 (GLUT4) protein concentration. CONCLUSION: Collectively, these findings suggest that chronic aerobic exercise enhances components of both the classical and novel insulin signalling cascades in normal rodent skeletal muscle, which may contribute to an increased insulin-stimulated plasma membrane GLUT4 protein concentration. PMID- 15799774 TI - Intramuscular pressure and tissue oxygenation during low-force static contraction do not underlie muscle fatigue. AB - AIM: To test the hypothesis that time-wise increase in intramuscular pressure (IMP) and subsequent decrease in muscle tissue oxygenation (TO(2)) results in muscle fatigue development during a non-exhaustive, low-force contraction evidenced by changes in electromyogram (EMG) and particular mechanomyogram (MMG). METHODS: Seven subjects performed static elbow flexion at 10% maximal voluntary contraction (MVC) for 10 min (10% MVC(10 min)). Surface EMG, MMG, IMP and TO(2) measured by near-infrared spectroscopy was recorded from m. biceps brachii during 10% MVC(10 min) and during 5% MVC test contractions of 1 min duration performed before 10% MVC(10 min), 10 and 30 min post-exercise. EMG and MMG were analysed for root mean square (rms) and mean power frequency (mpf). RESULTS: During 10% MVC(10 min) MMGrms increased from initial level of 0.04 +/- 0.01 to 0.11 +/- 0.07 m s(-2) in the last minute and MMGmpf and EMGmpf decreased from 34.9 +/- 8.2 to 21.3 +/- 3.8 Hz and from 71.7 +/- 10.9 to 61.7 +/- 10.0 Hz respectively. Similar changes were present in 5% MVC test contractions 30 min post-exercise. Initially, TO(2) decreased by 6.9 +/- 6.5% of resting level but returned to rest within 1 min. IMP remained constant during the contraction after an initial fourfold increase from resting level of 12.2 +/- 10.4 mmHg. CONCLUSIONS: IMP was anticipated to increase with time of contraction due to e.g. increased muscle water content; but this was not confirmed. Consequently, muscle blood flow was unlikely to be impeded with contraction time, which may account for the maintenance of TO(2). Thus, decreased TO(2) did not underlie either acute or long term muscle fatigue development evidenced by changes in EMG and particular MMG variables. PMID- 15799775 TI - Effects of cold-water immersion on VEGF mRNA and protein expression in heart and skeletal muscles of rats. AB - AIM: The effects of cold exposure on gene and protein expression of vascular endothelial growth factor (VEGF), in heart and skeletal muscles, were studied in male adult Wistar rats. METHODS: Cold immersion was accomplished by submerging the rats in shoulder-deep water maintained at approximately 18 degrees C, either acutely (1 h) or chronically (1 h day(-1), 5 days week(-1) for 20 weeks). The expressions of VEGF mRNA and protein in heart, gastrocnemius, and soleus muscles were examined by Northern and Western blotting and competitive-polymerase chain reaction techniques. RESULTS: The expressions of VEGF mRNA and protein were markedly increased in cardiac muscle of the cold-immersed group, particularly in the 1-hour exposure group, whereas VEGF mRNA and protein in gastrocnemius were decreased significantly after an acute exposure. Although the protein level in gastrocnemius remained low in the chronically exposed group, the expression of mRNA of VEGF(165) with chronic exposure in this group returned to the control level and that of VEGF(206) was 15% greater than that in controls. The expression of mRNA for VEGF(165) in soleus was also lowered by acute cold exposure, although that for VEGF(206) was stable. However, VEGF protein was increased by 50%. After 20 weeks, all of these parameters were increased over the levels found in the controls. CONCLUSION: These results suggest that the VEGF gene may be a major regulatory factor in cardiac and skeletal muscle adaptation to the cold environment stimulating angiogenesis and thermogenesis. PMID- 15799776 TI - The de novo methylation activity of Dnmt3a is distinctly different than that of Dnmt1. AB - BACKGROUND: Though Dnmt1 is considered the primary maintenance methyltransferase and Dnmt3a and Dnmt3b are considered de novo methyltransferases in mammals, these three enzymes may work together in maintaining as well as establishing DNA methylation patterns. It has been proposed that Dnmt1 may carry out de novo methylation at sites in the genome with transient single-stranded regions, such as replication origins, and then spread methylation from these nucleation sites in vivo, even though such activity has not been reported. RESULTS: In this study, we show that Dnmt3a does not act on single-stranded substrates in vitro, indicating that Dnmt3a is not likely to initiate DNA methylation at such proposed nucleation sites. Dnmt3a shows similar methylation activity on unmethylated and hemimethylated duplex DNA, though with some substrate preference. Unlike Dnmt1, pre-existing cytosine methylation at CpG sites or non-CpG sites does not stimulate Dnmt3a activity in vitro and in vivo. CONCLUSION: The fact that Dnmt3a does not act on single stranded DNA and is not stimulated by pre-existing cytosine methylation indicates that the de novo methylation activity of Dnmt3a is quite different from that of Dnmt1. These findings are consistent with a model in which Dnmt3a initiates methylation on one of the DNA strands of duplex DNA, and these hemimethylated sites then stimulate Dnmt1 activity for further methylation. PMID- 15799778 TI - Shooting darts: co-evolution and counter-adaptation in hermaphroditic snails. AB - BACKGROUND: Evolutionary conflicts of interest between the sexes often lead to co evolutionary arms races consisting of repeated arisal of traits advantageous for one sex but harmful to the other sex, and counter-adaptations by the latter. In hermaphrodites, these antagonistic interactions are at least an equally important driving force. Here, we investigate the evolution of one of the most striking examples of sexual conflict in hermaphrodites, the so-called shooting of love darts in land snails. Stabbing this calcareous dart through the partner's skin ultimately increases paternity. This trait is obviously beneficial for the shooter, but it manipulates sperm storage in the receiver. Hence, an arms race between the love-dart and the spermatophore receiving organs may be expected. RESULTS: We performed a detailed phylogenetic analysis of 28S ribosomal RNA gene sequences from dart-possessing land snail species. Both the Shimodaira-Hasegawa test and Bayesian posterior probabilities rejected a monophyletic origin of most reproductive structures, including the love-dart, indicating that most traits arose repeatedly. Based on the inferred phylogenetic trees, we calculated phylogenetically independent contrasts for the different reproductive traits. Subsequent principal component and correlation analyses demonstrated that these contrasts covary, meaning that correlated evolution of these traits occurred. CONCLUSION: Our study represents the first comprehensive comparative analysis of reproductive organ characteristics in simultaneous hermaphrodites. Moreover, it strongly suggests that co-evolutionary arms races can result from sexual conflict in these organisms and play a key role in the evolution of hermaphroditic mating systems. PMID- 15799777 TI - Floral gene resources from basal angiosperms for comparative genomics research. AB - BACKGROUND: The Floral Genome Project was initiated to bridge the genomic gap between the most broadly studied plant model systems. Arabidopsis and rice, although now completely sequenced and under intensive comparative genomic investigation, are separated by at least 125 million years of evolutionary time, and cannot in isolation provide a comprehensive perspective on structural and functional aspects of flowering plant genome dynamics. Here we discuss new genomic resources available to the scientific community, comprising cDNA libraries and Expressed Sequence Tag (EST) sequences for a suite of phylogenetically basal angiosperms specifically selected to bridge the evolutionary gaps between model plants and provide insights into gene content and genome structure in the earliest flowering plants. RESULTS: Random sequencing of cDNAs from representatives of phylogenetically important eudicot, non-grass monocot, and gymnosperm lineages has so far (as of 12/1/04) generated 70,514 ESTs and 48,170 assembled unigenes. Efficient sorting of EST sequences into putative gene families based on whole Arabidopsis/rice proteome comparison has permitted ready identification of cDNA clones for finished sequencing. Preliminarily, (i) proportions of functional categories among sequenced floral genes seem representative of the entire Arabidopsis transcriptome, (ii) many known floral gene homologues have been captured, and (iii) phylogenetic analyses of ESTs are providing new insights into the process of gene family evolution in relation to the origin and diversification of the angiosperms. CONCLUSION: Initial comparisons illustrate the utility of the EST data sets toward discovery of the basic floral transcriptome. These first findings also afford the opportunity to address a number of conspicuous evolutionary genomic questions, including reproductive organ transcriptome overlap between angiosperms and gymnosperms, genome-wide duplication history, lineage-specific gene duplication and functional divergence, and analyses of adaptive molecular evolution. Since not all genes in the floral transcriptome will be associated with flowering, these EST resources will also be of interest to plant scientists working on other functions, such as photosynthesis, signal transduction, and metabolic pathways. PMID- 15799779 TI - Mathematical model of blunt injury to the vascular wall via formation of rouleaux and changes in local hemodynamic and rheological factors. Implications for the mechanism of traumatic myocardial infarction. AB - BACKGROUND: Blood viscosity is fundamentally important in clinical practice yet the apparent viscosity at very low shear rates is not well understood. Various conditions such as blunt trauma may lead to the appearance of zones inside the vessel where shear stress equals zero. The aim of this research was to determine the blood viscosity and quantitative aspects of rouleau formation from erythrocytes at yield velocity (and therefore shear stress) equal to zero. Various fundamental differential equations and aspects of multiphase medium theory have been used. The equations were solved by a method of approximation. Experiments were conducted in an aerodynamic tube. RESULTS: The following were determined: (1) The dependence of the viscosity of a mixture on volume fraction during sedimentation of a group of particles (forming no aggregates), confirmed by published experimental data on the volume fractions of the second phase (f2) up to 0.6; (2) The dependence of the viscosity of the mixture on the volume fraction of erythrocytes during sedimentation of rouleaux when yield velocity is zero; (3) The increase in the viscosity of a mixture with an increasing erythrocyte concentration when yield velocity is zero; (4) The dependence of the quantity of rouleaux on shear stress (the higher the shear stress, the fewer the rouleaux) and on erythrocyte concentration (the more erythrocytes, the more rouleaux are formed). CONCLUSIONS: This work represents one of few attempts to estimate extreme values of viscosity at low shear rate. It may further our understanding of the mechanism of blunt trauma to the vessel wall and therefore of conditions such as traumatic acute myocardial infarction. Such estimates are also clinically significant, since abnormal values of blood viscosity have been observed in many pathological conditions such as traumatic crush syndrome, cancer, acute myocardial infarction and peripheral vascular disease. PMID- 15799780 TI - Effect of testosterone replacement or duration of castration on baroreflex bradycardia in conscious rats. AB - BACKGROUND: In this study, we tested the hypothesis that 17beta-estradiol contributes to testosterone-mediated restoration of baroreflex-mediated bradycardia in short-term (3 weeks) castrated rats. Further, a reported increase in serum testosterone after long-term (6 weeks) castration constituted a basis for testing the hypothesis that a spontaneous increase in serum testosterone or androstenedione in this model causes a commensurate increase in baroreflex mediated bradycardia. RESULTS: Testosterone (1 week) replacement enhanced baroreflex-mediated bradycardia in short-term castrated rats without changing 17beta-estradiol level. A spontaneous recovery of baroreflex-mediated bradycardia occurred following long-term castration, although circulating testosterone and androstenedione remained suppressed. CONCLUSION: The data suggest: 1) 17beta Estradiol does not contribute to testosterone restoration of the baroreflex mediated bradycardia in short-term castrated rats. 2) The long-term modulation of baroreflex-mediated bradycardia occurs independent of androgens, or the baroreflex mechanism may become adapted to low levels of circulating androgens. PMID- 15799781 TI - Hyperbolastic growth models: theory and application. AB - BACKGROUND: Mathematical models describing growth kinetics are very important for predicting many biological phenomena such as tumor volume, speed of disease progression, and determination of an optimal radiation and/or chemotherapy schedule. Growth models such as logistic, Gompertz, Richards, and Weibull have been extensively studied and applied to a wide range of medical and biological studies. We introduce a class of three and four parameter models called "hyperbolastic models" for accurately predicting and analyzing self-limited growth behavior that occurs e.g. in tumors. To illustrate the application and utility of these models and to gain a more complete understanding of them, we apply them to two sets of data considered in previously published literature. RESULTS: The results indicate that volumetric tumor growth follows the principle of hyperbolastic growth model type III, and in both applications at least one of the newly proposed models provides a better fit to the data than the classical models used for comparison. CONCLUSION: We have developed a new family of growth models that predict the volumetric growth behavior of multicellular tumor spheroids with a high degree of accuracy. We strongly believe that the family of hyperbolastic models can be a valuable predictive tool in many areas of biomedical and epidemiological research such as cancer or stem cell growth and infectious disease outbreaks. PMID- 15799783 TI - The GABBR1 locus and the G1465A variant is not associated with temporal lobe epilepsy preceded by febrile seizures. AB - BACKGROUND: Polymorphism G1465A in the GABBR1 gene has been suggested as a risk factor for non-lesional temporal lobe epilepsy (TLE); however, this genetic association study has not been independently replicated. We attempted to replicate this study in our cohort of patients with TLE. Furthermore, we also analyzed the coding sequence of this gene and searched for disease-causing mutations. METHODS: We included 120 unrelated individuals with TLE that was preceded by febrile seizures (FS) who did not have any evidence of structural lesions suggesting secondary epilepsy. 66 individuals had positive family history of TLE epilepsy and 54 were sporadic. Each patient was genotyped for the presence of G1465A polymorphism. All exons of the GABBR1 gene were screened by single strand confirmation polymorphism method. Genotypes were compared with two independent matched control groups. RESULTS: We detected two A alleles of the G1465A polymorphism in one homozygous control subject (0.87% of all alleles) and one A allele in a patient with TLE (0.45%, not significant). Other detected polymorphisms in coding regions had similar frequencies in epilepsy patients and control groups. No disease causing mutations in the GABBR1 gene were detected in patients with sporadic or familial TLE. CONCLUSION: Our results indicate that TLE preceded by FS is not associated with the polymorphisms or mutations in the GABBR1 gene, including the G1465A polymorphism. The proportion of TLE patients with FS in the original study, reporting this positive association, did not differ between allele A negative and positive cases. Thus, our failure to reproduce this result is likely applicable to all non-lesional TLE epilepsies. PMID- 15799784 TI - The measurement of response shift in patients with advanced prostate cancer and their partners. AB - BACKGROUND: There is increasing evidence to support the phenomenon of response shift (RS) in quality of life (QoL) studies, with many current QoL measures failing to allow for this. If significant response shift occurs amongst prostate cancer patients, it will be necessary to allow for this in the design of future clinical research and to reassess the conclusions of previous studies that have not allowed for this source of bias. This study therefore aimed to assess the presence of RS and psychosocial morbidity in patients with advanced prostate cancer and their partners. METHODS: 55 consecutive advanced prostate cancer patients and their partners completed the Prostate Cancer Patient & Partner questionnaire (PPP), shortly after diagnosis and again at 3 months and 6 months. At the follow-up visits, both patients and partners also completed a then-test in order to assess RS. RESULTS: Partners consistently showed greater psychological morbidity than patients in relation to the prostate cancer. This was most marked on the General Cancer Distress (GCD) subscale (p < 0.001, paired t-test), and regarding worries about treatment (p = 0.01). Significant RS was identified in partners and patients by the use of the then-test technique, particularly on the GCD subscale, the concerns about treatment and the concerns about urinary symptoms items. CONCLUSION: These results suggest the presence of RS in patients with advanced prostate cancer and their partners, with higher levels of psychosocial morbidity noted amongst partners. This is the first study to identify RS in partners and calls into question the interpretation of all studies assessing changes in QoL that fail to allow for this phenomenon. PMID- 15799785 TI - Correlation test to assess low-level processing of high-density oligonucleotide microarray data. AB - BACKGROUND: There are currently a number of competing techniques for low-level processing of oligonucleotide array data. The choice of technique has a profound effect on subsequent statistical analyses, but there is no method to assess whether a particular technique is appropriate for a specific data set, without reference to external data. RESULTS: We analyzed coregulation between genes in order to detect insufficient normalization between arrays, where coregulation is measured in terms of statistical correlation. In a large collection of genes, a random pair of genes should have on average zero correlation, hence allowing a correlation test. For all data sets that we evaluated, and the three most commonly used low-level processing procedures including MAS5, RMA and MBEI, the housekeeping-gene normalization failed the test. For a real clinical data set, RMA and MBEI showed significant correlation for absent genes. We also found that a second round of normalization on the probe set level improved normalization significantly throughout. CONCLUSION: Previous evaluation of low-level processing in the literature has been limited to artificial spike-in and mixture data sets. In the absence of a known gold-standard, the correlation criterion allows us to assess the appropriateness of low-level processing of a specific data set and the success of normalization for subsets of genes. PMID- 15799782 TI - MAPPER: a search engine for the computational identification of putative transcription factor binding sites in multiple genomes. AB - BACKGROUND: Cis-regulatory modules are combinations of regulatory elements occurring in close proximity to each other that control the spatial and temporal expression of genes. The ability to identify them in a genome-wide manner depends on the availability of accurate models and of search methods able to detect putative regulatory elements with enhanced sensitivity and specificity. RESULTS: We describe the implementation of a search method for putative transcription factor binding sites (TFBSs) based on hidden Markov models built from alignments of known sites. We built 1,079 models of TFBSs using experimentally determined sequence alignments of sites provided by the TRANSFAC and JASPAR databases and used them to scan sequences of the human, mouse, fly, worm and yeast genomes. In several cases tested the method identified correctly experimentally characterized sites, with better specificity and sensitivity than other similar computational methods. Moreover, a large-scale comparison using synthetic data showed that in the majority of cases our method performed significantly better than a nucleotide weight matrix-based method. CONCLUSION: The search engine, available at http://mapper.chip.org, allows the identification, visualization and selection of putative TFBSs occurring in the promoter or other regions of a gene from the human, mouse, fly, worm and yeast genomes. In addition it allows the user to upload a sequence to query and to build a model by supplying a multiple sequence alignment of binding sites for a transcription factor of interest. Due to its extensive database of models, powerful search engine and flexible interface, MAPPER represents an effective resource for the large-scale computational analysis of transcriptional regulation. PMID- 15799787 TI - A ring-like nucleoid is not necessary for radioresistance in the Deinococcaceae. AB - BACKGROUND: Transmission electron microscopy images of Deinococcus radiodurans R1 suggest that the nucleoid of this species exists as a "ring-like" body, and have led to speculation that this structure contributes to the radioresistance of the species. Since extreme radioresistance is characteristic of six other species of Deinococcus, we have attempted to correlate nucleoid morphology and radioresistance by determining whether the genomic DNA of each of these species exhibit similar structures. RESULTS: The nucleoid morphologies of seven recognized species of Deinococcus, the radioresistant bacterium Rubrobacter radiotolerans, and the more radiosensitive deinococcal relative Thermus aquaticus were evaluated using epifluorescence and deconvolution techniques. Although the nucleoids of Deinococcus murrayi, Deinococcus proteolyticus, Deinococcus radiophilus, and Deinococcus grandis have structures similar to D. radiodurans, the majority of nucleoids found in Deinococcus radiopugnans and Deinococcus geothermalis lack any specific organization. The nucleoid of R. radiotolerans consists of multiple highly condensed spheres of DNA scattered throughout the cell. The genomic DNA of Thermus aquaticus is uniformly distributed throughout the cell. CONCLUSION: There is no obvious relationship between the shape of a species' nucleoid and extreme radioresistance. However, the genomes of all extremely radioresistance species examined are highly condensed relative to more radiosensitive species. Whether DNA in this tightly packed configuration contributes to the radioresistance of these bacteria remains unknown, but this common structural feature appears to limit diffusion of fragments generated post irradiation even in cells incapable of repairing strand breaks. PMID- 15799789 TI - Microsurgical anatomy and approaches to the cavernous sinus. AB - OBJECTIVE: The aim of this article is to describe the anatomy of the cavernous sinus and to provide a guide for use when performing surgery in this complex area. Clinical cases are used to illustrate routes to the cavernous sinus and its contents and to demonstrate how the cavernous sinus can be used as a pathway for exposure of deeper structures. METHODS: Thirty cadaveric cavernous sinuses were examined using x3 to x40 magnification after the arteries and veins were injected with colored silicone. Distances between the entrance of the oculomotor and trochlear nerves and the posterior clinoid process were recorded. Stepwise dissections of the cavernous sinuses, performed to demonstrate the intradural and extradural routes, are accompanied by intraoperative photographs of those approaches. RESULTS: The anatomy of the cavernous sinus is complex because of the high density of critically important neural and vascular structures. Selective cases demonstrate how a detailed knowledge of cavernous sinus anatomy can provide for safer surgery with low morbidity. CONCLUSION: A precise understanding of the bony relationships and neurovascular contents of the cavernous sinus, together with the use of cranial base and microsurgical techniques, has allowed neurosurgeons to approach the cavernous sinus with reduced morbidity and mortality, changing the natural history of selected lesions in this region. Complete resection of cavernous sinus meningiomas has proven to be difficult and, in many cases, impossible without causing significant morbidity. However, surgical reduction of such lesions enhances the chances for success of subsequent therapy. PMID- 15799786 TI - The asthma epidemic and our artificial habitats. AB - BACKGROUND: The recent increase in childhood asthma has been a puzzling one. Recent views focus on the role of infection in the education of the immune system of young children. However, this so called hygiene hypothesis fails to answer some important questions about the current trends in asthma or to account for environmental influences that bear little relation to infection. DISCUSSION: The multi-factorial nature of asthma, reflecting the different ways we tend to interact with our environment, mandates that we look at the asthma epidemic from a broader perspective. Seemingly modern affluent lifestyles are placing us increasingly in static, artificial, microenvironments very different from the conditions prevailed for most part of our evolution and shaped our organisms. Changes that occurred during the second half of the 20th century in industrialized nations with the spread of central heating/conditioning, building insulation, hygiene, TV/PC/games, manufactured food, indoor entertainment, cars, medical care, and sedentary lifestyles all seem to be depriving our children from the essential inputs needed to develop normal airway function (resistance). Asthma according to this view is a manifestation of our respiratory maladaptation to modern lifestyles, or in other words to our increasingly artificial habitats. The basis of the artificial habitat notion may lie in reduced exposure of innate immunity to a variety of environmental stimuli, infectious and non-infectious, leading to reduced formulation of regulatory cells/cytokines as well as inscribed regulatory pathways. This could contribute to a faulty checking mechanism of non functional Th2 (and likely Th1) responses, resulting in asthma and other immuno dysregulation disorders. SUMMARY: In this piece I discuss the artificial habitat concept, its correspondence with epidemiological data of asthma and allergy, and provide possible immunological underpinning for it from an evolutionary perspective of health and disease. PMID- 15799788 TI - Polymorphism screening and haplotype analysis of the tryptophan hydroxylase gene (TPH1) and association with bipolar affective disorder in Taiwan. AB - BACKGROUND: Disturbances in serotonin neurotransmission are implicated in the etiology of many psychiatric disorders, including bipolar affective disorder (BPD). The tryptophan hydroxylase gene (TPH), which codes for the enzyme catalyzing the rate-limiting step in serotonin biosynthetic pathway, is one of the leading candidate genes for psychiatric and behavioral disorders. In a preliminary study, we found that TPH1 intron7 A218C polymorphism was associated with BPD. This study was designed to investigate sequence variants of the TPH1 gene in Taiwanese and to test whether the TPH1 gene is a susceptibility factor for the BPD. METHODS: Using a systematic approach, we have searched the exons and promoter region of the TPH1 gene for sequence variants in Taiwanese Han and have identified five variants, A-1067G, G-347T, T3804A, C27224T, and A27237G. These five variants plus another five taken from the literature and a public database were examined for an association in 108 BPD patients and 103 controls; no association was detected for any of the 10 variants. RESULTS: Haplotype constructions using these 10 SNPs showed that the 3 most common haplotypes in both patients and controls were identical. One of the fourth common haplotype in the patient group (i.e. GGGAGACCCA) was unique and showed a trend of significance with the disease (P = 0.028). However, the significance was abolished after Bonferroni correction thus suggesting the association is weak. In addition, three haplotype-tagged SNPs (htSNPs) were selected to represent all haplotypes with frequencies larger than 2% in the Taiwanese Han population. The defined TPH1 htSNPs significantly reduce the marker number for haplotype analysis thus provides useful information for future association studies in our population. CONCLUSION: Results of this study did not support the role of TPH1 gene in BPD etiology. As the current studies found the TPH1 gene under investigation belongs to the peripheral serotonin system and may link to a cardiac dysfunction phenotype, a second TPH gene that functions predominantly in the brain (i.e., nTPH or TPH2) should be the target for the future association study. PMID- 15799790 TI - Exposure of midline cranial base without a facial incision through a combined craniofacial-transfacial procedure. AB - OBJECTIVE: A single-stage combined craniofacial-transfacial approach that exposes the midline cranial base without visible facial incisions is described. METHODS: Between 1992 and 1998, eight patients underwent surgery for five different anterior cranial base pathological findings: four angiofibromas, one mesenchymal chondrosarcoma, one esthesioneuroblastoma, one odontogenic myxoma, and one encephalocele. In all cases, the surgical exposure consisted of a bicoronal scalp incision with a bifrontal craniotomy and fronto-orbitonasal osteotomy, and then a sublabial incision for transmaxillary exposure. RESULTS: Gross total resection was achieved in five cases. The encephalocele was resected with complete reconstruction of the bony defect. Seven patients developed complications, primarily wound infections, cerebrospinal fluid leaks, and anemia. Postoperative Karnofsky Performance Scale scores ranged between 80 and 100 (mean, 92.5). Long term follow-up information (mean, 56 mo; median, 59.5 mo; range, 5-108 mo) was available for all patients. CONCLUSION: Large anterior cranial base lesions can be resected and excellent cosmetic outcomes can be achieved with a single-stage combined transfacial-craniofacial approach that exposes the entire midline cranial base and requires no facial incisions. PMID- 15799791 TI - Lateral ventricle tumors: surgical strategies according to tumor origin and development--a series of 72 cases. AB - OBJECTIVE: Optimal surgical management in lateral ventricle tumors remains controversial. We conducted a retrospective study of patients with these lesions treated with a surgical strategy on the basis of tumor origin: primary or secondary ventricular and associated transependymal development. METHODS: A total of 72 patients underwent surgery for lateral ventricle tumors. The mean patient age was 39 years (range, 6 mo to 78 yr). Raised intracranial pressure occurred in 53% of patients, followed by mental disturbances or psychiatric symptoms (32%) and motor deficits (21%). The transcortical approach was used in 44 patients, and an interhemispheric approach was used in 28 patients; a transcallosal approach was used in 16 patients, and a parasplenial approach was used in 12 patients. Neuropsychological tests were performed in selected patients. RESULTS: Total resection was performed in 82% of patients. Sixty-five percent of tumors were benign and low-grade tumors. There was no surgical mortality, and the morbidity rate was 11%. Postoperative epilepsy (5.9%) was significantly increased in the transcortical group. The mean follow-up period was 55 months; 59% of patients achieved good recovery and moderate disability. In postoperative neuropsychological testing sessions, deficits in verbal memory were observed in six patients (8%). Final morbidity correlated well with preoperative clinical condition and pathological diagnosis. CONCLUSION: Lateral ventricle tumors can be treated best by careful selection of the approach according to tumor origin and development. Overall, the transcallosal approach is preferred, but in patients with transependymal growth or large primary or secondary ventricular tumors, the transcortical is a better option. PMID- 15799792 TI - Surgical technique of lumbar artificial disc replacement with the Charite artificial disc. AB - Lumbar artificial disc technology has been commercially available outside the United States for nearly 2 decades. With the Food and Drug Administration approval of the Charite Artificial Disc in October 2004, an entirely new spinal surgeon population will be able to offer this technology to their patients as a treatment option. As with other techniques in spinal surgery, indications for lumbar total disc replacement are paramount to the success of the procedure. The correct surgical technique is also important to a successful outcome. This article describes the technique for placement of the Charite Artificial Disc in indicated patients. The technique is similar to that of an anterior lumbar interbody fusion procedure, but many differences between the techniques make lumbar total disc replacement a unique procedure in the spinal surgeon's armamentarium. Although this article is thorough in its description of the surgical technique for total disc replacement with the Charite Artificial Disc, it should not be used as a substitute for company-sponsored training. PMID- 15799793 TI - Cervical arthroplasty with the Bryan disc. AB - Cervical arthroplasty is a promising nonfusion alternative for the treatment of degenerative disc disease. After anterior cervical discectomy for neurological decompression, the intervertebral space is reconstructed by use of a metal and polymer prosthesis, allowing semiconstrained motion in multiple planes. This approach allows for preservation of cervical motion, potentially reducing the risk of transitional-level disease. PMID- 15799794 TI - Reconstruction options for complex middle cerebral artery aneurysms. AB - OBJECTIVE: To describe techniques of reconstruction for unclippable and uncoilable middle cerebral artery aneurysms. METHODS: A retrospective review was performed of seven patients who underwent eight operations during a 9-year period to treat complex middle cerebral artery bifurcation aneurysms not amenable to direct clipping or endovascular coiling. All preoperative and postoperative clinical and imaging data were reviewed. Follow-up was obtained for all patients via clinic visit and/or telephone. RESULTS: The operative techniques used included saphenous vein graft bypass (n = 1), radial artery graft interposition (n = 2), radial artery patch (n = 1), superficial temporal artery interposition graft (n = 1), superior thyroid artery interposition graft (n = 1), direct reimplantation of branch (n = 1), and reconstruction of trifurcation (n = 1). There was no mortality. Six patients had excellent outcomes with Glasgow Outcome Scale scores of 5, and one patient had a good outcome with a Glasgow Outcome Scale score of 4. CONCLUSION: Techniques for middle cerebral artery reconstruction may remain important and useful in the age of endovascular aneurysm treatment. PMID- 15799795 TI - The cut flow index: an intraoperative predictor of the success of extracranial intracranial bypass for occlusive cerebrovascular disease. AB - OBJECTIVE: There has been a resurgence of interest in selective extracranial intracranial bypass for revascularization of cerebrovascular occlusive disease. We evaluated the usefulness of intraoperative blood flow measurements in predicting graft success after extracranial-intracranial bypass. METHODS: A retrospective review of 51 cases of extracranial-intracranial bypass for purposes of flow augmentation in the setting of cerebrovascular occlusive disease was performed. In all cases, free flow from the cut end of the donor vessel, termed cut flow, was measured. The cut flow index (CFI) (bypass flow [ml/min] / cut flow [ml/min]) was derived and correlated with bypass patency, postoperative bypass flow, cerebrovascular reserve testing, and clinical outcome. RESULTS: The CFI was a significant predictor of bypass patency (P = 0.002). Using a CFI of 0.5 as a threshold, the bypass patency rate was 92% in cases with a CFI greater than 0.5 compared with 50% in cases with a CFI less than 0.5. Intraoperative bypass flow correlated well with postoperative measurements obtained from quantitative phase contrast magnetic resonance imaging. An analysis of cases with a poor CFI indicated that a logical interpretation of bypass function can be performed during surgery. CONCLUSION: A poor CFI can alert surgeons to potential difficulties with the donor vessel, anastomosis, or recipient vessel during surgery. Furthermore, a CFI closely approximating 1.0 provides physiological confirmation of impaired cerebrovascular reserve in the recipient bed. PMID- 15799796 TI - Cortical surface tracking using a stereoscopic operating microscope. AB - OBJECTIVE: To measure and compensate for soft tissue deformation during image guided neurosurgery, we have developed a novel approach to estimate the three dimensional (3-D) topology of the cortical surface and track its motion over time. METHODS: We use stereopsis to estimate the 3-D cortical topology during neurosurgical procedures. To facilitate this process, two charge-coupled device cameras have been attached to the binocular optics of a stereoscopic operating microscope. Before surgery, this stereo imaging system is calibrated to obtain the extrinsic and intrinsic camera parameters. During surgery, the 3-D shape of the cortical surface is automatically estimated from a stereo pair of images and registered to the preoperative image volume to provide navigational guidance. This estimation requires robust matching of features between the images, which, when combined with the camera calibration, yields the desired 3-D coordinates. After the 3-D cortical surface has been estimated from stereo pairs, its motion is tracked by comparing the current surface with its previous locations. RESULTS: We are able to estimate the 3-D topology of the cortical surface with an average error of less than 1.2 mm. Executing on a 1.1-GHz Pentium machine, the 3-D estimation from a stereo pair of 1024 x 768 resolution images requires approximately 60 seconds of computation. By applying stereopsis over time, we are able to track the motion of the cortical surface, including the pulsatile movement of the cortical surface, gravitational sag, tissue bulge as a result of increased intracranial pressure, and the parenchymal shape changes associated with tissue resection. The results from 10 surgical patients are reported. CONCLUSION: We have demonstrated that a stereo vision system coupled to the operating microscope can be used to efficiently estimate the dynamic topology of the cortical surface during surgery. The 3-D surface can be coregistered to the preoperative image volume. This unique intraoperative imaging technique expands the capability of the current navigational system in the operating room and increases the accuracy of anatomic correspondence with preoperative images through compensation for brain deformation. PMID- 15799797 TI - Functional identification of the primary motor area by corticospinal tractography. AB - OBJECTIVE: For quick and stable identification of the primary motor area (PMA), diffusion tensor imaging (DTI) data were acquired and corticospinal tractography was mathematically visualized. METHODS: Data sets of DTI, anatomic magnetic resonance imaging, and functional magnetic resonance imaging with finger-tapping tasks were acquired during the same investigation in 30 patients with a brain lesion affecting the motor system. Off-line processing of DTI data was performed to visualize the corticospinal tract, placing a seed area in the cerebral peduncle of the midbrain, where the corticospinal tract is densely concentrated. Somatosensory evoked magnetic fields and intraoperative cortical somatosensory evoked potentials were recorded with electrical stimulation of the median nerve to confirm the results of the corticospinal tractography. RESULTS: Functional magnetic resonance imaging and somatosensory evoked magnetic fields failed to identify the PMA in eight patients (16.7%) and one patient (3.8%) investigated, respectively, because of cortical dysfunctions caused by brain lesions. DTI data were acquired within 3 minutes without patient tasks. Using the appropriate seed area and fractional anisotropy, corticospinal tractography successfully indicated the PMA location in all patients. The suspected PMA and central sulcus locations were confirmed by the cortical somatosensory evoked potentials. CONCLUSION: Corticospinal tractography enables identification of the PMA and is beneficial, particularly for patients who present with dysfunction of the PMA. PMID- 15799798 TI - Frameless stereotaxy to facilitate anterolateral thoracolumbar surgery: technique. AB - With the advent of frameless stereotaxy and its application to the spine, more precise and less invasive spinal procedures are possible. In addition to being less invasive, these techniques may increase surgeon confidence and allow shorter operating times. Described here is a case of Pott's disease of the thoracolumbar spine and how intraoperative image guidance can facilitate operative progress and accuracy in a patient in whom the underlying disease has severely deformed the normal anatomy of the spine. Added confidence about the location of vital structures as the surgeon proceeds with resection of the vertebral bodies and discs is depicted. Facilitation with image-guided placement of bicortical vertebral body screws and an interbody device is demonstrated. A diagram of the recommended positioning of the equipment in the operating room is provided along with "pearls" learned from our experience with this application. We believe that even the most experienced and skilled surgeon will find facilitation of anterior thoracolumbar surgery with image guidance to be of considerable benefit. PMID- 15799799 TI - Bulbar compression by an ectatic vertebral artery: a novel neurovascular construct relieved by microsurgical decompression. AB - OBJECTIVE: Brainstem compression caused by vascular abnormalities has rarely been reported in the literature. We describe five cases of large ectatic vertebral artery causing compression and distortion of the medulla oblongata with pyramidal tract signs and low cranial nerve dysfunction. Microvascular decompression by retracting the vertebral artery and anchoring it to the dura has been the treatment of choice. METHODS: Five patients, four male and one female, presented with progressive myelopathic features and lower cranial nerve dysfunction, especially dysphonia and dysphagia. Four patients were affected by systemic arterial hypertension. Magnetic resonance imaging showed impingement of the right vertebral artery in three patients and the left vertebral artery in two patients, on the right and left lateral medulla, respectively. In two patients, hypoplasia of the contralateral vertebral artery was documented. RESULTS: All patients underwent neurovascular decompression of the medulla oblongata. The ectatic and tortuous vertebral artery was detached from the medulla, shifted away, and repositioned by anchoring to the nearby dura mater using a Gore-Tex vascular slip. Postoperatively, all patients but one had improvement of their previous neurological symptoms. CONCLUSION: Brainstem dysfunction caused by a tortuous ectatic vertebral artery might be less uncommon than expected. It should be considered a new distinct clinical entity, the real incidence of which needs to be carefully evaluated by an appropriate diagnostic protocol, which includes primarily magnetic resonance imaging with specific three-dimensional sequences. Awareness of this condition is necessary to ensure the appropriate treatment. Surgical microvascular decompression seems very effective. PMID- 15799800 TI - Microsurgical removal of intraventricular lesions using endoscopic visualization and stereotactic guidance. AB - OBJECTIVE: To demonstrate the technique of stereotactic microsurgical endoscopic removal of intraventricular tumors or colloid cysts assisted by intraoperative computed tomography. METHODS: We adapted a tubular "ventriculoport" for stereotactic insertion of an endoscope into the ventricle. This facilitated microsurgical resection of 14 intraventricular tumors or colloid cysts by use of intraoperative stereotactic microsurgical endoscopic removal of intraventricular tumors or colloid cysts assisted by intraoperative computed tomography. RESULTS: Gross total resection was achieved in 12 patients and confirmed by intraoperative computed tomographic scanning and postoperative magnetic resonance imaging. Patients with preoperative hydrocephalus had relief of their symptoms. Perioperative morbidity was limited to mild headache associated with postoperative pneumocephalus. The average length of stay was 3.6 days. Twelve patients had significant improvement in their symptoms. CONCLUSION: The combination of intraoperative computed tomography-guided stereotactic technique and rigid endoscopy facilitated an accurate, minimally invasive, microsurgical removal of these intraventricular masses. This approach minimized retraction and provided satisfactory visualization. PMID- 15799801 TI - Sequential visualization of brain and fiber tract deformation during intracranial surgery with three-dimensional ultrasound: an approach to evaluate the effect of brain shift. AB - OBJECTIVE: We present a technique that allows intraoperative display of brain shift and its effects on fiber tracts. METHODS: Three patients had intracranial lesions (one malignant glioma, one metastasis, and one cavernoma) in contact with either the corticospinal or the geniculostriate tract that were removed microneurosurgically. Preoperatively, magnetic resonance diffusion-weighted imaging (DWI) was performed to visualize the fiber tract at risk. DWI data were fused with those obtained from anatomic T1-weighted magnetic resonance imaging. A single-rack three-dimensional ultrasound neuronavigation system, which simultaneously displays the MRI scan and the corresponding ultrasound image, was used intraoperatively for 1) navigation; 2) definition of fixed and potentially shifting ultrasound landmarks near the fiber tract; and 3) sequential image updating at different steps of resection. The result was time-dependent brain deformation data. With a standard personal computer equipped with standard image software, the brain shift-associated fiber tract deformation was assessed by use of sequential landmark registration. After surgery, DWI was performed to confirm the predicted fiber tract deformation. RESULTS: The lesions were removed without morbidity. Comparison of three-dimensional ultrasound with DWI and T1-weighted magnetic resonance imaging data allowed us to define fixed and potentially shifting landmarks close to the respective fiber tract. Postoperative DWI confirmed that the actual fiber tract position at the conclusion of surgery corresponded to the sonographically predicted fiber tract position. CONCLUSION: By definition and sequential intraoperative registration of ultrasound landmarks near the fiber tract, brain shift-associated deformation of a tract that is not visible sonographically can be assessed correctly. This approach seems to help identify and avoid eloquent brain areas during intracranial surgery. PMID- 15799802 TI - Waterjet dissection versus ultrasonic aspiration in epilepsy surgery. AB - OBJECTIVE: Waterjet dissection is currently under close investigation in neurosurgery. Experimentally, precise brain parenchyma dissection with vessel preservation has been demonstrated. Clinically, the safety of the instrument has already been proved. However, precise data demonstrating that waterjet dissection indeed reduces surgical blood loss are still missing. METHODS: The authors applied the waterjet device in a prospective randomized study in comparison with the ultrasonic aspirator. Because there is little variability in the procedure, 30 patients with temporal lobe epilepsy receiving a tailored temporal lobe resection between December 1999 and October 2002 were selected for this study. Intraoperative vessel preservation, intraoperative blood loss, surgical complications, and epilepsy outcome were evaluated. All patients were followed at 3-month intervals. RESULTS: During surgery, both instruments were easy to handle. Only with the waterjet dissector, however, were even small intraparenchymal blood vessels preserved. Intraoperative blood loss was significantly reduced with the waterjet (mean, 70 +/- 46 ml) compared with the ultrasonic aspirator (mean, 121 +/- 48 ml). However, no difference in the necessity for blood transfusion occurred. No difference was observed with respect to operation time (238.6 +/- 37.0 min with the waterjet, 247.5 +/- 41.5 min with the ultrasonic aspirator), surgical complications, and outcome. CONCLUSION: The waterjet dissector enables a significant reduction of intraoperative blood loss in the investigated setting. However, further studies are needed to confirm these results with a larger number of patients. Studies also are needed to prove that the reduction of blood loss is of clinical relevance for the outcome of the patients. PMID- 15799803 TI - Using Dermabond for wound closure in lumbar and cervical neurosurgical procedures. AB - OBJECTIVE: 2-Octylcyanoacrylate (Dermabond; Ethicon, Inc., Somerville, NJ) is a liquid adhesive being used with increasing frequency for the closure of lacerations and surgical incisions. Dermabond provides excellent cosmetic closure, and recent studies have demonstrated very low infection risks when it is properly applied. There are no published studies using Dermabond on lumbar or cervical procedures. This study was undertaken to determine whether Dermabond is safe and efficacious to use in these common neurosurgical procedures. METHODS: Records of 200 consecutive patients with Dermabond closure after anterior cervical discectomy, microlumbar discectomy, or lumbar laminectomy by the senior author (JEB) with a mean follow-up time of 5.42 months were retrospectively reviewed. Suspected infections with or without confirmatory cultures, erythema, and incisional drainage were documented. RESULTS: Of 200 patients, 85 underwent microlumbar discectomy, 22 lumbar laminectomy, and 93 anterior cervical discectomy. There was only 1 definitive infection, which was a culture-proven discitis in a microlumbar discectomy patient. Of the remaining 85 microlumbar discectomies, there was 1 transient incisional erythema. Of the 22 lumbar laminectomies, there was 1 patient with clinical superficial wound infection with negative cultures and 4 patients with transient incisional drainage without infection. Of the 93 anterior cervical discectomies, 2 had transient incisional drainage without infection. CONCLUSION: This study demonstrates that Dermabond is safe to use in neurosurgery patients undergoing lumbar or cervical procedures, with only 1 patient of 200 having a proven infection. Patients are able to shower and do not have sutures or staples to remove. Patient responses are overwhelmingly positive. PMID- 15799804 TI - Biomechanical comparison of two stabilization techniques of the atlantoaxial joints: transarticular screw fixation versus screw and rod fixation. AB - OBJECTIVE: To compare the biomechanical stability imparted to the C1 and C2 vertebrae by either transarticular screw fixation (TSF) or screw and rod fixation (SRF) techniques in a cadaver model. METHODS: Ten fresh ligamentous human cervical spine specimens were harvested from cadavers. The specimens were tested sequentially in the intact state, after injury and stabilization (unilateral left side and bilateral), and after fatiguing to 5000 cycles (0.5 Hz) at +/-1.0 N.m of flexion and extension. The specimens were stabilized by use of TSF in 5 spines or SRF in the other 5 spines. The data were converted to angular displacements, and the stabilized cases were compared with intact states for evaluating the efficacies of the two techniques in stabilizing the C1-C2 segments. RESULTS: In the TSF group, the unilateral fixation using one screw imparted a significant stability in only the axial rotation mode. The unilateral procedure in the SRF group was effective in stabilization in all modes except in extension. The bilateral procedure in both of the groups was effective across the C1-C2 segment. However, the SRF group afforded higher stability than the corresponding TSF group in the flexion and extension modes. The degree of stability did not change after fatigue compared with the prefatigue data. CONCLUSION: In general, a surgeon should undertake a bilateral fixation to achieve sufficient stability across the atlantoaxial complex, and either technique will provide satisfactory results, although the SRF technique may be better in the flexion and extension modes. One should use the SRF procedure while trying to achieve stability with a unilateral system. PMID- 15799805 TI - Landmarks for vertebral artery repositioning in bulbar compression syndrome: anatomic and microsurgical nuances. AB - OBJECTIVE: The purpose of this study was to better elucidate the anatomic relationship between the vertebral artery (VA) along with its perforating vessels and the brainstem to develop anatomic guidelines that would be helpful when decompressing medulla oblongata compressed by the VA. METHODS: Microanatomy dissection was performed in six formalin-fixed cadaveric heads. The VA, posteroinferior cerebellar artery (PICA), anteroinferior cerebellar artery (AICA), and lower brainstem perforators were examined under magnification using a surgical microscope. The outer diameters of the VA, PICA, and AICA were measured. The distance between the VA, lying within the lateral cerebellomedullary cistern, and the medulla oblongata was quantified. The lower brainstem perforating vessels were examined in relation to their course, outer diameter, and length. RESULTS: In four of six brains, the left VA was dominant, presenting an outer diameter at least 0.8 mm larger than the contralateral VA. The average distance between the VA and the medulla oblongata was 3 mm. The perforating branches presented a mean outer diameter of 0.7 mm. In particular, perforators arising from the VA and PICA (Groups 1 and 2) presented an average outer diameter of 0.2 mm and an average length of 7 mm. Perforating vessels arising from the AICA and vertebrobasilar junction (Groups 3 and 4) presented an average outer diameter of 0.4 mm and an average length of 12 mm. CONCLUSION: This work provides new information that may be useful to minimize the risk of injury of perforators when operating on medulla oblongata compression by the VA. PMID- 15799806 TI - Surgical exposure of the sciatic nerve in the gluteal region: anatomic and historical comparison of two approaches. AB - OBJECTIVE: To increase awareness among neurosurgeons of alternative surgical approaches to lesions of the sciatic nerve in the gluteal region. METHODS: The dominant surgical approach to lesions of the proximal sciatic nerve involves detachment and medial reflection of the gluteus maximus through a question-mark incision. An alternative to this infragluteal exposure is a transgluteal approach, which provides access to the sciatic nerve by splitting the gluteus maximus through a curvilinear incision. We explored the anatomy and surgical history of these approaches through cadaveric study, our own case series, and a literature review. RESULTS: The infragluteal approach uses a larger incision, extensive dissection, and postoperative bracing while allowing wide exposure of the nerve inferiorly into the thigh. By contrast, the transgluteal approach minimizes dissection and spares muscle attachments but requires meticulous attention to hemostasis and provides a more focal exposure of the sciatic nerve. During the past century, the infragluteal approach has been described more frequently and has become increasingly popular among peripheral-nerve surgeons. For comparison, we present three patients in whom the transgluteal approach was used to treat substantial lesions of the proximal sciatic nerve. CONCLUSION: At the present time, the majority of peripheral nerve surgeons use an infragluteal approach to the proximal sciatic nerve. However, for select patients with well defined and localized lesions, the transgluteal approach may provide sufficient nerve exposure with lowered operative complexity and postoperative morbidity. PMID- 15799807 TI - Orbitozygomatic approach to basilar apex aneurysms. AB - Basilar apex region aneurysms are among the most complex cerebral aneurysms. They are not, however, among the most common aneurysms, and increased use of endovascular treatment has further decreased the number of patients with these lesions who undergo surgery. Nonetheless, not all basilar apex aneurysms are amenable to coil embolization, and neurosurgeons must be prepared to treat patients with basilar apex aneurysms surgically. We prefer an orbitozygomatic craniotomy and transsylvian approach. Meticulous exercise of the basic tenets of aneurysm surgery (proximal vascular control, sharp dissection, and preservation of perforating vessels) is crucial to optimal patient outcomes. PMID- 15799808 TI - Anatomic and surgical basis of the sphenoid ridge keyhole approach for cerebral aneurysms. AB - In vascular neurosurgery, the pterional approach has been used primarily for the treatment of a wide variety of diseases (cavernous angiomas, arteriovenous malformations, etc.), and it is used to take advantage of naturally occurring planes and spaces to expose the major structures of the circle of Willis. It provides access to the major part of the anterior circulation aneurysms and those occurring in the upper and most proximal part of the posterior circulation. Conversely, there has been an increasing interest in the so-called minimally invasive procedures or keyhole approaches to treating cerebral aneurysms in specific locations. In this work, we describe a novel keyhole approach that was conceived to achieve the angle of vision and advantages of the classic pterional approach. This surgical approach is based on the anatomic location of the sphenoid ridge and its relationship with the sylvian fissure and basal cisterns. The initial incision is made over the hairline behind the external border of the eye on the side selected. A skin and muscular flap is reflected anteriorly, and a small 3 x 3-cm craniotomy is completed around the external landmarks of the sphenoid ridge. Further extradural drilling is completed down to the anterior clinoid process. The dura is opened in a semilunar manner, and the sylvian fissure is opened completely to reach the sylvian and basal cisterns. Thereafter, the aneurysm is dissected and clipped according to the standard microtechnique of the neurosurgeon. A step-by-step description of the approach is offered in this work to facilitate a clear understanding of it. We recommend this approach for treatment of aneurysms arising at the anterior part of the circle of Willis. It has the advantages of less operative time, fewer days of hospitalization, and similar morbidity and mortality compared with the standard pterional craniotomy (5.7% on our service for nongiant ruptured aneurysms). PMID- 15799809 TI - Superficial temporal artery-to-middle cerebral artery bypass. AB - The superficial temporal artery-to-middle cerebral artery bypass represents the mainstay of bypass operations for cerebral revascularization. The role of this operation in the setting of cerebrovascular occlusive disease, with compromised cerebral hemodynamic reserve, is a topic of current investigation. In this report, we describe technical nuances to avert potential pitfalls in the performance of this procedure. The use of intraoperative quantitative blood flow measurements as a reliable method of assessing the success of the bypass is also described. PMID- 15799810 TI - Techniques for the ventral correction of postsurgical cervical kyphotic deformity. AB - Progressive kyphotic deformity of the cervical spine may be a late sequela of congenital cervical anomalies, degenerative disc disease, neoplasm, cervical trauma, and surgical procedures. Of these, postsurgical kyphosis is the most common and can occur after ventral and dorsal surgeries of the cervical spine. The purpose of this article is to review the causes and clinical presentation of postsurgical cervical kyphosis and to focus on the operative planning and ventral correction techniques. PMID- 15799811 TI - Ideal suture diameter is critical for consistent middle cerebral artery occlusion in mice. AB - OBJECTIVE: The use of transgenic and knockout mice has led to a need for a consistent model of mouse transient focal cerebral ischemia. In a great majority of the published mouse middle cerebral artery (MCA) occlusion studies, the methods indicated the type of intraluminal suture used without indicating the actual suture diameter after modification. We attempted to determine the ideal suture diameter to produce consistent occlusion in the MCA of adult male C57BL/6 mice. METHODS: Suture tips were coated to a depth 4 mm with glue, and 6-0 sutures of eight different, precisely measured diameters were produced. The coated 6-0 sutures in different diameters were introduced 10 mm into the internal carotid artery via the external carotid artery of the mice to produce MCA occlusion (n = 40; five animals for each diameter), and the mice (22-24 g) were transaortically perfused with saline. The base of the brain was exposed, and photographs of the vessels were obtained before and after transaortic injection of Evans blue dye to determine the consistency of MCA occlusion for each suture diameter. Cerebral blood flow was measured 10 minutes before occlusion and 20 minutes after reperfusion, and 2,3,5-triphenyltetrazolium chloride staining was performed to demonstrate the ischemic damage in additional animals with 110-microm (n = 5) and 180-microm (n = 8) diameter sutures. RESULTS: Sutures measuring 170 microm and 180 microm in diameter consistently occluded the MCA of C57BL/6 mice. In addition, 2,3,5-triphenyltetrazolium chloride staining demonstrated consistent infarction with 180-microm diameter sutures. The infarct volume was 36.3 +/- 4.2 mm3. CONCLUSION: Small changes in the diameter of the occlusion suture tip affect consistency in the mouse MCA occlusion model. PMID- 15799812 TI - Awake craniotomy for microsurgical obliteration of mycotic aneurysms: technical report of three cases. AB - OBJECTIVE AND IMPORTANCE: Infectious (mycotic) aneurysms that do not resolve with medical treatment require surgical obliteration, usually requiring sacrifice of the parent artery. In addition, patients with mycotic aneurysms frequently need subsequent cardiac valve repair, which often necessitates anticoagulation. Three cases of awake craniotomy for microsurgical clipping of mycotic aneurysms are presented. Awake minimally invasive craniotomy using frameless stereotactic guidance on the basis of computed tomographic angiography enables temporary occlusion of the parent artery with neurological assessment before obliteration of the aneurysm. CLINICAL PRESENTATION: A 56-year-old woman presented with progressively worsening mitral valve disease and a history of subacute bacterial endocarditis and subarachnoid hemorrhage 30 years previously. A cerebral angiogram revealed a 4-mm left middle cerebral artery (MCA) angular branch aneurysm, which required obliteration before mitral valve replacement. The second patient, a 64-year-old woman with a history of rheumatic fever, had an 8-mm right distal MCA aneurysm diagnosed in the setting of pulmonary abscess and worsening cardiac function as a result of mitral valve disease. The third patient, a 57 year-old man with a history of fevers, night sweats, and progressive mitral valve disease, had an enlarging left MCA angular branch aneurysm despite the administration of antibiotics. Because of their location on distal MCA branches, none of the aneurysms were amenable to preoperative test balloon occlusion. INTERVENTION: After undergoing stereotactic computed tomographic angiography with fiducial markers, the patients underwent a minimally invasive awake craniotomy with frameless stereotactic navigation. In all cases, the results of the neurological examination were unchanged during temporary parent artery occlusion and the aneurysms were successfully obliterated. CONCLUSION: Awake minimally invasive craniotomy for an infectious aneurysm located in eloquent brain enables awake testing before permanent clipping or vessel sacrifice. Combining frameless stereotactic navigation with computed tomographic angiography allowed us to perform the operation quickly through a small craniotomy with minimal exploration. PMID- 15799813 TI - Use of "bonnet" bypass with radial artery interposition graft in a patient with recurrent cranial base carcinoma: technical report of two cases and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Two patients with recurrent cranial base carcinomas involving the carotid artery received a "bonnet" bypass using the contralateral superficial temporal artery as the donor vessel because the ipsilateral common and external carotid arteries were unavailable. The radial artery was used as the graft. CLINICAL PRESENTATION: A 58-year-old man with ear pain and an enlarging mass involving the left cranial base and neck had undergone a right partial glossectomy and modified neck dissection followed by radiotherapy for squamous cell carcinoma. Recurrent carcinoma extensively involved the left internal carotid artery. A 46-year-old man with jaw pain and hoarseness had undergone multiple resections and radiation therapy for medullary thyroid carcinoma. Magnetic resonance imaging showed recurrent tumor on the right. INTERVENTION: Both patients underwent a pterional craniotomy. The supraclinoid internal carotid artery was exposed. The radial artery was harvested. The contralateral superficial temporal artery was dissected at its bifurcation into the frontal and parietal branches. The radial artery graft was anastomosed to the superficial temporal artery and a recipient ipsilateral branch of the middle cerebral artery. The internal carotid artery was clip-ligated. After surgery, both patients remained neurologically stable. Angiography confirmed that the bypasses were patent and that the middle cerebral artery territory filled. The patients' carcinomas were resected aggressively. CONCLUSION: When aggressive resection of cranial base tumors is needed and the ipsilateral carotid artery is unavailable as a donor vessel, a "bonnet" bypass with carotid artery sacrifice may be performed. Compared with vein grafts, microsurgical anastomosis is easier and the patency rate is higher with a radial artery graft. PMID- 15799814 TI - Traumatic atlanto-occipital dislocation: presentation of a new posterior occipitoatlantoaxial fixation technique in an adult survivor: technical case report. AB - OBJECTIVE AND IMPORTANCE: More survivors of traumatic atlanto-occipital dislocation (AOD) in adults have recently been reported. Surgical management options are therefore of increasing interest. We present a new technique of posterior C0-C1-C2 fixation. CLINICAL PRESENTATION: A 29-year-old motorcyclist survived a traumatic vertical AOD of 15 mm. No spinal cord or medullary lesion was present. Brain contusion and diffuse axonal injuries led to a cortical biplegia, which recovered progressively over a period of 6 months. Twelve months after surgery, no neurological deficit was present except for slightly increased deep tendon reflexes. INTERVENTION: Posterior C0-C1-C2 fixation was performed with two bicortical occipital screws, one bicortical lateral mass screw in the atlas, and one monocortical pars screw in the axis on each side, connected to a plate-rod on the right and left sides. Fusion was performed with monocortical bone graft from the posterior iliac crest. CONCLUSION: The surgical technique described was thought to be safe to perform and resulted in immediate stability without external immobilization. Solid fusion was achieved 6 months after surgery. PMID- 15799815 TI - Pars screw fixation of a hangman's fracture: technical case report. AB - OBJECTIVE AND IMPORTANCE: Traumatic spondylolisthesis of the axis may be treated by external immobilization or surgical fixation. CLINICAL PRESENTATION: We report the case of a 23-year-old man who sustained an Effendi Type II fracture of the axis, for which halo immobilization did not provide adequate stability. INTERVENTION: The unstable fracture was treated by placing lag screws in the pars interarticularis of C2, which reduced the fracture directly but sacrificed no normal spinal motion. The patient developed a solid fusion, and cervical alignment was normal at his 6-month follow-up examination. CONCLUSION: Although this technique has been reported previously, it is more commonly used in multilevel cervical fusions than for stand-alone repair of C2. Management options, anatomy, and technical considerations for the treatment of traumatic spondylolisthesis are reviewed. PMID- 15799816 TI - Cervicothoracic junction arthroplasty after previous fusion surgery for adjacent segment degeneration: case report. AB - OBJECTIVE AND IMPORTANCE: This is the first reported case of cervical arthroplasty using the Bryan Cervical Disc Prosthesis System (Medtronic Sofamor Danek, Inc., Memphis, TN) in the management of adjacent segment degeneration associated with previous fusion surgery and surgery at the cervicothoracic junction. CLINICAL PRESENTATION: This case report describes a 25-year-old woman who initially underwent a two-level anterior cervical fusion in 1998, 2 years after being involved in a motor vehicle accident. She was well until 18 months before presentation, when she developed bilateral shoulder pain, mechanical neck pain worse on flexion, and bilateral C8 distribution arm pain and paresthesia. On clinical examination, no focal deficits were found, although the range of motion was reduced. Preoperative cervical spine x-rays and magnetic resonance scanning confirmed accelerated degeneration of the C4-C5 and C7-T1 disc spaces, with evidence of neural compression at those levels. INTERVENTION: After careful consideration of various treatment options and failure of all conservative measures, the patient underwent an anterior C4-C5 and C7-T1 decompression with removal of the anterior cervical plate and placement of two artificial disc prostheses. After surgery, her course was uncomplicated and she was discharged from hospital well. There was complete resolution of the arm symptoms and reduction of the neck pain, with a reduction in the amount of analgesia she was taking. Seven months after surgery, she remains well with repeat x-rays confirming motion at the operated levels. CONCLUSION: This case demonstrates that cervical arthroplasty is a reasonable treatment option for patients who have had previous surgery in which interbody fusion has been performed and who have developed degeneration of adjacent levels. Despite the altered biomechanics at the cervicothoracic junction, no adverse features were noted with arthroplasty at this level. PMID- 15799817 TI - Endoscopic aqueductal stenting via the fourth ventricle under navigating system guidance: technical note. AB - OBJECTIVE: We report three patients with symptomatic isolated fourth ventricle after ventriculoperitoneal shunt placement for hydrocephalus associated with ventricular hemorrhage. All three patients were treated successfully with our new method of endoscopic aqueductal stenting under navigating system guidance. METHODS: A therapeutic rigid endoscope was inserted through the thin cerebellar hemisphere, and endoscopic aqueductal stenting was performed via the enlarged fourth ventricle under navigating system guidance. RESULTS: All three patients underwent successful procedures with good outcomes. CONCLUSION: Our method of aqueductal stenting is a reasonable choice for initial treatment of patients with isolated fourth ventricle, and it entails less invasive neurosurgery. PMID- 15799818 TI - Titanium cage-assisted polymethylmethacrylate reconstruction for cervical spinal metastasis: technical note. AB - OBJECTIVE: Reconstruction and stabilization of the cervical spine after vertebrectomy is an important goal in the surgical management of spinal metastasis. The authors describe their reconstruction technique using a titanium cage-Silastic tube construct injected with polymethylmethacrylate (PMMA) augmented by an anterior cervical plate. The surgical results using this technique are reviewed. METHODS: Six patients ranging from 43 to 70 years of age underwent resection of metastatic tumor in the cervical spine followed by cage assisted PMMA reconstruction of the anterior spinal column. The following reconstruction technique was performed. A Silastic tube is incised longitudinally and placed circumferentially around a titanium cage with the opening facing anteriorly. The cage-Silastic tube construct is carefully tapped into the corpectomy defect and filled with PMMA. The final construct is then augmented with anterior cervical plate fixation. RESULTS: Two patients required additional posterior stabilization with lateral mass screws and rods. All patients achieved immediate stabilization, restoration of vertebral body height and normal lordosis, and preservation of the ability to walk independently. Five patients experienced significant palliation of biomechanical neck pain. There were no complications of neurological worsening, postoperative hematoma, wound infection, subsidence, graft dislodgement, or construct failure during a follow-up period of 1 to 19 months (mean, 6.8 mo). CONCLUSION: Titanium cage-assisted PMMA reconstruction augmented with an anterior cervical plate is an effective means of reconstruction after tumor resection in patients with cervical spinal metastasis. The Silastic tube holds the PMMA within the cage and protects the spinal cord from potential thermal injury. PMID- 15799819 TI - Epigenetic silencing of the protocadherin family member PCDH-gamma-A11 in astrocytomas. AB - In a microarray-based methylation analysis of astrocytomas [World Health Organization (WHO) grade II], we identified a CpG island within the first exon of the protocadherin-gamma subfamily A11 (PCDH-gamma-A11) gene that showed hypermethylation compared to normal brain tissue. Bisulfite sequencing and combined bisulfite restriction analysis (COBRA) was performed to screen low- and high-grade astrocytomas for the methylation status of this CpG island. Hypermethylation was detected in 30 of 34 (88%) astrocytomas (WHO grades II and III), 20 of 23 (87%) glioblastomas (WHO grade IV), and 8 of 8 (100%) glioma cell lines. There was a highly significant correlation (P = .00028) between PCDH-gamma A11 hypermethylation and decreased transcription as determined by competitive reverse transcription polymerase chain reaction in WHO grades II and III astrocytomas. After treatment of glioma cell lines with a demethylating agent, transcription of PCDH-gamma-A11 was restored. In summary, we have identified PCDH gamma-A11 as a new target silenced epigenetically in astrocytic gliomas. The inactivation of this cell-cell contact molecule might be involved in the invasive growth of astrocytoma cells into normal brain parenchyma. PMID- 15799820 TI - Clinical implications of promoter hypermethylation in RASSF1A and MGMT in retinoblastoma. AB - We investigated the epigenetic silencing and genetic changes of the RAS associated domain family 1A (RASSF1A) gene and the O6-methylguanine-DNA methyltransferase (MGMT) gene in retinoblastoma. We extracted DNA from microdissected tumor and normal retina tissues of the same patient in 68 retinoblastoma cases. Promoter methylation in RASSF1A and MGMT was analyzed by methylation-specific PCR, RASSF1A sequence alterations in all coding exons by direct DNA sequencing, and RASSF1A expression by RT-PCR. Cell cycle staging was analyzed by flow cytometry. We detected RASSF1A promoter hypermethylation in 82% of retinoblastoma, in tumor tissues only but not in adjacent normal retinal tissue cells. There was no expression of RASSF1A transcripts in all hypermethylated samples, but RASSF1A transcripts were restored after 5-aza-2' deoxycytidine treatment with no changes in cell cycle or apoptosis. No mutation in the RASSF1A sequence was found. MGMT hypermethylation was present in 15% of the retinoblastoma samples, and the absence of MGMT hypermethylation was associated (P = .002) with retinoblastoma at advanced Reese-Ellsworth tumor stage. Our results revealed a high RASSF1A hypermethylation frequency in retinoblastoma. The correlation of MGMT inactivation by promoter hypermethylation with lower-stage diseases indicated that MGMT hypermethylation provides useful prognostic information. Epigenetic mechanism plays an important role in the progression of retinoblastoma. PMID- 15799821 TI - Bone microenvironment modulates expression and activity of cathepsin B in prostate cancer. AB - Prostate cancers metastasize to bone leading to osteolysis. Here we assessed proteolysis of DQ-collagen I (a bone matrix protein) and, for comparison, DQ collagen IV, by living human prostate carcinoma cells in vitro. Both collagens were degraded, and this degradation was reduced by inhibitors of matrix metallo, serine, and cysteine proteases. Because secretion of the cysteine protease cathepsin B is increased in human breast fibroblasts grown on collagen I gels, we analyzed cathepsin B levels and secretion in prostate cells grown on collagen I gels. Levels and secretion were increased only in DU145 cells--cells that expressed the highest baseline levels of cathepsin B. Secretion of cathepsin B was also elevated in DU145 cells grown in vitro on human bone fragments. We further investigated the effect of the bone microenvironment on cathepsin B expression and activity in vivo in a SCID-human model of prostate bone metastasis. High levels of cathepsin B protein and activity were found in DU145, PC3, and LNCaP bone tumors, although the PC3 and LNCaP cells had exhibited low cathepsin B expression in vitro. Our results suggest that tumor-stromal interactions in the context of the bone microenvironment can modulate the expression of the cysteine protease cathepsin B. PMID- 15799822 TI - The role of heparanase in lymph node metastatic dissemination: dynamic contrast enhanced MRI of Eb lymphoma in mice. AB - Heparanase expression has been linked to increased tumor invasion, metastasis, and angiogenesis and with poor prognosis. The aim of the study was to monitor the effect of heparanase expression on lymph node metastasis, in heparanase overexpressing subcutaneous Eb mouse T-lymphoma tumors, and their draining lymph node. Dynamic contrast-enhanced magnetic resonance imaging (MRI) using biotin-BSA GdDTPA-FAM/ROX was applied for analysis of blood volume, vascular permeability, and interstitial convection, and for detection of very early stages of such metastatic dissemination. Eb tumors increased extravasation, interstitial convection, and lymphatic drain of the contrast material. Interstitial flow directions were mapped by showing radial outflow interrupted in some tumors by directional flow toward the popliteal lymph node. Heparanase expression significantly increased contrast enhancement of the popliteal lymph node but not of the primary tumor. Changes in MR contrast enhancement preceded the formation of pathologically detectable metastases, and were detectable when only a few enhanced green fluorescent protein (EGFP)-expressing Eb cells were found near and within the nodes. These results demonstrate very early, heparanase-dependent vascular changes in lymph nodes that were visible by MRI following administration of biotin-BSA-GdDTPA-FAM/ROX, and can be used for studying the initial stages of lymph node infiltration. PMID- 15799823 TI - Bone marrow-derived lin(-)c-kit(+)Sca-1+ stem cells do not contribute to vasculogenesis in Lewis lung carcinoma. AB - The development of tumor vasculature is thought to occur through two complementary processes: sprouting angiogenesis from preexisting blood vessels of the host, and vasculogenesis, which involves the spontaneous development of vessels through specific recruitment, differentiation, and vascular incorporation of circulating endothelial cells (EC), endothelial progenitor cells (EPC), or potentially bone marrow-derived cells. Recent reports, however, have challenged the belief that bone marrow-derived cells contribute to tumor neovascularization, claiming an exclusive role for sprouting angiogenesis in tumor blood vessel development. In the present study, we explored the recruitment behavior of bone marrow-derived lin(-)c-kit(+)Sca-1+ stem cells to subcutaneously implanted Lewis lung carcinoma in a syngeneic bone marrow transplantation model. We observed that although lin(-)c-kit(+)Sca-1+ and their derived cells demonstrate significant recruitment to carcinomas in vivo, they do not appear to functionally contribute to tumor neovascularization. Furthermore, our results support the hypothesis that new vessel formation in carcinomas occurs primarily through endothelialization from adjacent and preexisting vasculature. PMID- 15799824 TI - Prostate-specific antigen modulates the expression of genes involved in prostate tumor growth. AB - Prostate-specific antigen (PSA) is a serine protease that is widely used as a surrogate marker in the early diagnosis and management of prostate cancer. The physiological relevance of tissue PSA levels and their role in prostate tumor growth and metastasis are not known. Free-PSA (f-PSA) was purified to homogeneity from human seminal plasma by column chromatography, eliminating hk2 and all known PSA complexes and retaining its protease activity. Confluent monolayers of prostate cancer cell lines, PC-3M and LNCaP, were treated with f-PSA in a series of in vitro experiments to determine the changes in expression of various genes that are known to regulate tumor growth and metastasis. Gene array, quantitative polymerase chain reaction (QPCR), and enzyme-linked immunosorbent assay (ELISA) results show significant changes in the expression of various cancer-related genes in PC-3M and LNCaP cells treated with f-PSA. In a gene array analysis of PC 3M cells treated with 10 muM f-PSA, 136 genes were upregulated and 137 genes were downregulated. In LNCaP cells treated with an identical concentration of f-PSA, a total of 793 genes was regulated. QPCR analysis reveals that the genes for urokinase-type plasminogen activator (uPA), VEGF, and Pim-1 oncogene, known to promote tumor growth, were significantly downregulated, whereas IFN-gamma, known to be a tumor-suppressor gene, was significantly upregulated in f-PSA-treated PC 3M cells. The effect of f-PSA on VEGF and IFN-gamma gene expression and on protein release in PC-3M cells was distinctly dose-dependent. In vivo studies showed a significant reduction (P = .03) in tumor load when f-PSA was administered in the tumor vicinity of PC-3M tumor-bearing BALB/c nude mice. Our data support the hypothesis that f-PSA plays a significant role in prostate tumor growth by regulating various proangiogenic and antiangiogenic growth factors. PMID- 15799825 TI - Antisense-mediated suppression of Heparanase gene inhibits melanoma cell invasion. AB - Cancer metastasis, is a frequent manifestation of malignant melanoma progression. Successful invasion into distant organs by tumor cells must include attachment to microvessel endothelial cells, and degradation of basement membranes and extracellular matrix (ECM). Heparan sulfate proteoglycans (HSPG) are essential and ubiquitous macromolecules associated with the cell surface and ECM of a wide range of cells and tissues. Heparanase (HPSE-1) is an ECM degradative enzyme, which degrades the heparan sulfate (HS) chains of HSPG at specific intrachain sites. To investigate effects of changes in heparanase gene expression in metastatic melanoma cells, we constructed adenoviral vectors containing the full length human HPSE-1 cDNA in both sense (Ad-S/hep) and antisense orientations (Ad AS/hep). We found increased HPSE-1 expression and activity in melanoma cell lines following Ad-S/hep infection by Western blot analyses and specific HPSE-1 activity assay. Conversely, HPSE-1 content was significantly inhibited following infection with Ad-AS/Hep. Importantly, HPSE-1 modulation by these adenoviral constructs correlated with invasive cellular properties in vitro and in vivo. Our results suggest that HPSE-1 not only contributes to the invasive phenotype of melanoma cells, but also that the Ad-AS/hep-mediated inhibition of its enzymatic activity can be efficacious in the prevention and treatment of melanoma metastasis. PMID- 15799826 TI - Utilizing optical tomography with ultrasound localization to image heterogeneous hemoglobin distribution in large breast cancers. AB - PURPOSE: Angiogenesis in advanced breast cancers is highly distorted and heterogeneous. Noninvasive imaging that can monitor angiogenesis may be invaluable initially for diagnosis and then for assessing tumor response to treatment. By combining ultrasound (US) and near-infrared (NIR) optical imaging, a reliable new technique has emerged for localizing and characterizing tumor angiogenesis within the breast. METHODS: This new technique employs a commercial US transducer coupled with an array of NIR optical fibers mounted on a hand-held probe. The US image is used for lesion localization and for guiding optical imaging reconstruction. Optical sensors are used for imaging tumor total hemoglobin distribution, which is directly related to tumor angiogenesis. RESULTS: Six large breast carcinomas were studied and microvessel density count was then performed on tissue samples obtained from these cancers. Two patients had locally advanced breast cancers and received neoadjuvant chemotherapy for 3 months. In one patient, before chemotherapy, the total hemoglobin distribution showed a high concentration at the cancer periphery; the distribution was later confined to the core area after 3 months of treatment. In another patient, as treatment progressed, the maximum hemoglobin concentration decreased from 255.3, to 147.5, to 76.9 micromol/l with an associated reduction in spatial extension. The other four patients had cancers of 2.0 to 3.0 cm in size and were imaged either at the time of core biopsy or definitive surgery. The histologic microvessel density counts from these tumor samples correlate to hemoglobin distributions with a correlation coefficient of 0.64 (P < .05). CONCLUSION: These initial results suggest that this new imaging technique may have great potential in imaging the heterogeneous vascular distribution of larger breast cancers in vivo and in monitoring treatment-related changes in angiogenesis during chemotherapy. PMID- 15799827 TI - Integrin alpha v beta 3-targeted imaging of lung cancer. AB - A series of radiolabeled cyclic arginine-glycine-aspartic acid (RGD) peptide ligands for cell adhesion molecule integrin alpha v beta 3-targeted tumor angiogenesis targeting are being developed in our laboratory. In this study, this effort continues by applying a positron emitter 64Cu-labeled PEGylated dimeric RGD peptide radiotracer 64Cu-DOTA-PEG-E[c(RGDyK)]2 for lung cancer imaging. The PEGylated RGD peptide indicated integrin alpha v beta 3 avidity, but the PEGylation reduced the receptor binding affinity of this ligand compared to the unmodified RGD dimer. The radiotracer revealed rapid blood clearance and predominant renal clearance route. The minimum nonspecific activity accumulation in normal lung tissue and heart rendered high-quality orthotopic lung cancer tumor images, enabling clear demarcation of both the primary tumor at the upper lobe of the left lung, as well as metastases in the mediastinum, contralateral lung, and diaphragm. As a comparison, fluorodeoxyglucose (FDG) scans on the same mice were only able to identify the primary tumor, with the metastatic lesions masked by intense cardiac uptake and high lung background. 64Cu-DOTA-PEG E[c(RGDyK)]2 is an excellent position emission tomography (PET) tracer for integrin-positive tumor imaging. Further studies to improve the receptor binding affinity of the tracer and subsequently to increase the magnitude of tumor uptake without comprising the favorable in vivo kinetics are currently in progress. PMID- 15799837 TI - Social Security... or Social Sick-urity? PMID- 15799828 TI - Inverse relationship between 15-lipoxygenase-2 and PPAR-gamma gene expression in normal epithelia compared with tumor epithelia. AB - 15-Lipoxygenase-2 (15-LOX-2) synthesizes 15-S-hydroxyeicosatetraenoic acid (15-S HETE), an endogenous ligand for the nuclear receptor, peroxisome proliferator activated receptor-gamma (PPAR-gamma). Several studies have described an inverse relationship between 15-LOX-2 and PPAR-gamma expression in normal versus tumor samples. To systematically determine if this is a ubiquitous phenomenon, we used a variety of epithelial and nonepithelial cells and some tissues to further evaluate the extent of this inverse relationship. The levels of mRNA or protein were measured by reverse transcriptase polymerase chain reaction or Western gray level intensity, whereas distribution was determined by in situ hybridization or immunofluorescence. 15-S-HETE was measured by liquid chromatography/tandem mass spectrometry. Normal epithelial cells/samples generally expressed high levels of 15-LOX-2 along with the enzyme product 15-S-HETE, but both levels were reduced in cancer cells/samples. In contrast, most cancer cells expressed high levels of PPAR-gamma mRNA and protein, which were absent from normal epithelial cells. Overall, the inverse relationship between these two genes was primarily restricted to epithelial samples. Forced expression of PPAR-gamma reduced 15-LOX 2 protein levels in normal cells, whereas forced expression of 15-LOX-2 in tumor cells suppressed PPAR-gamma protein levels. These results suggest that feedback mechanisms may contribute to the loss of 15-LOX-2 pathway components, which coincide with an increase in PPAR-gamma in many epithelial cancers. PMID- 15799838 TI - Healthy workplace environments support a nurturing student educational climate. AB - Due to the shortage of medical laboratory scientist personnel in the United States, the successful integration of students into the Medical Laboratory Science (MLS) profession is vital. Education Facilitators are being used increasingly to assist clinical site managers in actively supporting and contributing to a healthier workplace environment, thereby inspiring students to become part of the organization. Education Facilitators associated with a single university-based MLS Program were the participant target group in a study. Face to-face, in-depth, semi-structured interviews were conducted with five Education Facilitators at two clinical sites. The findings of this study suggest that organizational efforts to improve the learning environment for MLS students should focus on providing a supportive and nurturing educational climate. Education Facilitators' descriptions revealed an organizational leadership that communicated a commitment to provide people, space, and equipment in an effort to promote a positive clinical rotation experience. Additionally, the facilitators expressed a need to "pay more attention to students," be "willing to work with students," and make sure the students don't get "lost in the shuffle." They said that concern for a healthy work/life balance should be communicated to MLS students through day-to-day educational contact with bench technologists and Education Facilitators. PMID- 15799829 TI - PPAR-gamma activation inhibits angiogenesis by blocking ELR+CXC chemokine production in non-small cell lung cancer. AB - Activation of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) results in inhibition of tumor growth in various types of cancers, but the mechanism(s) by which PPAR-gamma induces growth arrest has not been completely defined. In a recent study, we demonstrate that treatment of A549 (human non small cell lung cancer cell line) tumor-bearing SCID mice with PPAR-gamma ligands troglitazone (Tro) and pioglitazone significantly inhibits primary tumor growth. In this study, immunohistochemical analysis of Tro-treated and Pio-treated tumors with factor VIII antibody revealed a significant reduction in blood vessel density compared to tumors in control animals, suggesting inhibition of angiogenesis. Further analysis showed that treatment of A549 cells in vitro with Tro or transient transfection of A549 cells with constitutively active PPAR-gamma (VP16-PPAR-gamma) construct blocked the production of the angiogenic ELR+CXC chemokines IL-8 (CXCL8), ENA-78 (CXCL5), and Gro-alpha (CXCL1). Similarly, an inhibitor of NF-kappa B activation (PDTC) also blocked CXCL8, CXCL5, and CXCL1 production, consistent with their NF-kappa B-dependent regulation. Conditioned media from A549 cells induce human microvascular endothelial cell (HMVEC) chemotaxis. However, conditioned media from Tro-treated A549 cells induced significantly less HMVEC chemotaxis compared to untreated A549 cells. Furthermore, PPAR-gamma activation inhibited NF-kappa B transcriptional activity, as assessed by TransAM reporter gene assay. Collectively, our data suggest that PPAR-gamma ligands can inhibit tumor-associated angiogenesis by blocking the production of ELR+CXC chemokines, which is mediated through antagonizing NF kappaB activation. These antiangiogenic effects likely contribute to the inhibition of primary tumor growth by PPAR-gamma ligands. PMID- 15799839 TI - Direct access testing: nuts and bolts of starting DAT in a hospital environment. AB - Direct Access Testing (DAT) is a consumer-driven phenomenon that is sweeping the country. It is a highly controversial subject, the implementation of which requires methodical planning and development to be successful. DAT's purpose is to allow the consumer to order a battery of laboratory tests previously available only with a physician's order. With proper planning, DAT can be beneficial to a medical facility in a number of ways. Whether introduced to a hospital or an integrated health system, physician buy-in is vital to the success of DAT. Physicians need to publicly champion the program, even if members of the laboratory are doing the behind-the-scenes work. DAT is growing rapidly, but everyone still wants the stamp of approval from their physician(s) when it comes to their health. PMID- 15799840 TI - Using data to make decisions and drive results: a LEAN implementation strategy. AB - During the process of facility planning, Fairview Laboratory Services utilized LEAN manufacturing to maximize efficiency, simplify processes, and improve laboratory support of patient care services. By incorporating the LEAN program's concepts in our pilot program, we were able to reduce turnaround time by 50%, improve productivity by greater than 40%, reduce costs by 31%, save more than 440 square feet of space, standardize work practices, reduce errors and error potential, continuously measure performance, eliminate excess unused inventory and visual noise, and cross-train 100% of staff in the core laboratory. In addition, we trained a core team of people that is available to coordinate future LEAN projects in the laboratory and other areas of the organization. PMID- 15799841 TI - Building the culture of quality: you have the tools in your laboratory. PMID- 15799842 TI - Blowing the whistle on poor laboratory performances, part 2. To report or to not report. PMID- 15799843 TI - How to write manuals to fulfill CLIA requirements. PMID- 15799844 TI - [Effect of the Helicobacter pylori eradication in patients with functional dyspepsia: randomised placebo-controlled trial]. AB - BACKGROUND AND OBJECTIVE: The Helicobacter pylori eradication in patients with functional dyspepsia has been the subject of controversy because trials come to contradictory conclusions. The objective of this trial was to evaluate the effect, compared with placebo, of the eradication treatment in patients with functional dyspepsia. PATIENTS AND METHOD: Randomized double blind placebo controlled trial. We included 158 patients attended by family physicians (Area 10 Primary Care, Health Institute of Madrid) with functional dyspepsia and Helicobacter pylori infection detected by the ureasa test in endoscopy. An OCA (ameprazole clarithromycin and amoxicillin group (n = 79) and a placebo group (n = 79) were randomized. During 7 days, patients at the OCA group received omeprazole (20 mg bid), clarithromycin (500 mg bid) and amoxicillin (1000 mg bid) daily, and patients at the control group received the placebo agent twice daily. Dyspepsia improvement according to a Likert scale (5 steps), and eradication of H. pylori by 13C-urea breath test were evaluated during one year. RESULTS: Both groups were homogeneous in relation to age, sex and dyspepsia degree. The average age (standard deviation) of studied patients was 41.99 (13.93) years. At one year of follow up, H. pylori was eradicated in 81.01% (64/79) of the OCA group and 5.06% (4/79) of the placebo group. The difference of dyspepsia improvement (22.78%; 95% confidence interval [CI], 7.62-37.79) between the OCA group (41.77%; 95% CI, 30.77-53.41), and the placebo group (18.99%; 95% CI 11.03-29.38) was significant (p = 0.0018). CONCLUSION: Eradication of Helicobacter pylori in patients with functional dyspepsia is more effective improving symptoms than placebo. PMID- 15799845 TI - [PREVENCAT study: control of cardiovascular risk in primary care]. AB - BACKGROUND AND OBJECTIVE: Most studies of cardiovascular risk factors (CVRF) conducted in our environment concentrate in a single CVRF. The PREVENCAT study was designed to estimate the control of CVRF in the population attended in primary care presenting arterial hypertension (HT), type 2 diabetes mellitus (DM2) and/or hypercholesterolemia (HC) as well as to assess the prevalence of Metabolic Syndrome in these patients. PATIENTS AND METHOD: Multicenter, cross sectional study, in patients with HT, DM2 and/or HC, consecutively recruited by primary care physicians in Spain. The blood pressure, cholesterol, basal glycaemia, obesity, smoking and physical activity were assessed. The degree of control of these CVRF and the prevalence of MS were estimated. RESULTS: 2,649 patients were recruited, aged 64 (11.3) years, with a 51.6% of women. The most frequent diagnosis was HT (78.9%), followed by HC (58.4%) and DM2 (37.4%). In the whole sample, the percentages of patients who had a control or had initially normal values of blood pressure, cholesterol and basal glycemia were 40.0% (confidence interval [CI], 95% 38.2-41.9), 42.6% (95% CI, 40.5-44.7) and 62.7% (95% CI, 60.8-64.5), respectively. 15.6% of cases (95% CI, 14.3-17.0) had body mass index < or = 25 kg/m2; 87.5% were non-current smokers (95% CI, 86.2-88.8); and 46.2% practiced regular physical activity (95% CI, 44.3-48.1). 40% of patients had < or = 2 CVRF in good control. The prevalence of metabolic syndrome was 50.6% (95% CI, 48.7-52.5). CONCLUSIONS: The control of the CVRF considered in primary care attended population is insufficient. Hardly one of each 2 patients with HT, DM2 and HC is under control. The overweight and sedentarism control is still poorer. PMID- 15799846 TI - [Probability of developing and dying of cancer in Catalonia during the period 1998-2001]. AB - BACKGROUND AND OBJECTIVE: We intended to estimate the probability of developing and dying from cancer in Catalonia during the period 1998-2001. PATIENTS AND METHOD: We used a Bayesian model which incorporates data from the Tarragona and Girona Cancer Registries and from the Catalonia Mortality Registry. The probability of developing and dying from cancer has been calculated using a competitive risk-based methodology. RESULTS: Lifetime probability of developing cancer in Catalonia is almost 1 out of 2 (43.7%) for men and 1 out of 3 (32.1%) in women. The probability of dying from cancer is 29.1% for men and 17.9% in women. 67% of men and 56% of women diagnosed with cancer will die from this disease. One out of 14 men will develop a lung cancer during his life and 1 out of 11 women will develop breast cancer. CONCLUSIONS: The observed rising in the probability of developing cancer in Catalonia over the last ten years highlights even more than ever the importance of this health problem. PMID- 15799847 TI - [Control of cardiovascular risk factors in primary healthcare: do we control the factors or the risk?]. PMID- 15799848 TI - [DNA polymorphisms and alcoholism]. PMID- 15799849 TI - [Reactive Airways Dysfunction Syndrome. A study of 18 cases]. AB - BACKGROUND AND OBJECTIVE: We intended to perform a descriptive-evolutive study of a series of patients diagnosed with Reactive Airways Dysfunction Syndrome. PATIENTS AND METHOD: Retrospective study of 18 patients diagnosed with Reactive Airways Dysfunction Syndrome. Clinical data, functional respiratory study, and allergic and laboratory tests were collected. The clinical, functional and occupational evolution one year after diagnosis was evaluated. RESULTS: Twelve men and 4 women aged 27 to 66 years were evaluated. More frequent activities were metallurgy and cleaning, and the related agents were derived from chlorine and sulphur. Time from the beginning of exposure to development of symptoms was 1 hour in 10 (55%) patients and less than 24 hours in all cases. All patients reported to have dyspnea (100%), 12 (67%) wheezing, 8 (44%) nose and throat symptoms, 8 (44%) general symptoms such as headache and vomits, and 4 (22%) ocular manifestations. Although all patients received medical assistance after exposure, a correct diagnosis was made some months or years later (average time 3.6 years). At that time there was an airway obstruction in 6 of 18 (33%) patients. With regard to the clinical evolution, there was an improvement in 8 of 18 (44%) patients, while the rest remained stable. In the functional follow up, 8 of 17 (47%) showed no changes, 6 of 17 (35%) improved and 3 of 17 (18%) worsened. Among the patients who inhaled in the workplace, 7 of 14 (50%) continued working in the same job, 5 of 14 (36%) were off sick and 2 of 14 (14%) changed their role in the same company. CONCLUSIONS: Reactive Airways Dysfunction Syndrome is not frequently suspected and thus its diagnosis and treatment can be delayed for months or years. PMID- 15799850 TI - [Development of endocrinological systemic diseases in the adult, the fetal programming hypothesis]. PMID- 15799851 TI - [Role of endoscopic capsule in the diagnosis of digestive diseases]. AB - The recent development of the endoscopic capsule allows for the endoscopic study of the whole small intestine on a safe and non-invasive basis. Digestive hemorrhage of unknown origin is the main indication of the technique (sensitivity 89%, specificity 95%). In these cases, a high diagnostic accuracy is seen in patients with active bleeding (92.3%), which nevertheless decreases notably in patients with previous non-active bleeding (12.9%). Capsule endoscopy has also been proposed in patients with a clinical and laboratory suspicion of Crohn's disease involving the small bowel which has not been diagnosed through conventional techniques. Finally, the usefulness of capsule endoscopy for the management of other diseases such as celiac disease, intestinal tumors, gastrointestinal polyposis, abdominal pain of unknown origin, in pediatric patients or in the bowel involvement of systemic diseases should be established by means of well-designed, broad studies. PMID- 15799852 TI - [Glucose metabolic disorders in relation with insulin resistance in pediatric age]. PMID- 15799853 TI - [Evaluation of a guideline for Jehova's witnesses in a surgical practice]. PMID- 15799854 TI - [Cholinesterase as a marker in acute septic conditions]. PMID- 15799855 TI - [Study of malnutrition in hospitalized elderly patients]. PMID- 15799856 TI - [Prevalence of thyroid dysfunction in the elderly]. PMID- 15799857 TI - [Treatment of choroidal melanoma. Study of 13 cases]. PMID- 15799858 TI - [Migraine but not epilepsy is the commonest neurological disease during pregnancy]. PMID- 15799859 TI - [Additions to the CONSORT statement]. PMID- 15799860 TI - Efficacy of lamivudine in the treatment of children with chronic hepatitis B. AB - The major aim of our study was the assessment of the efficacy of lamivudine therapy in children with chronic hepatitis B (CHB). One hundred and six children with CHB, from 5- to 17-years old, were administered lamivudine 3mg/kg/day, up to 100mg/day, for 48 weeks. To assess the efficacy of the therapy several biochemical and virological parameters were monitored before, during, at the end of treatment and 24 weeks after therapy. The applied therapy resulted in a statistically significant decrease of ALT activity and HBV replication within the studied group. Biochemical and virological responses to anti-viral therapy were simultaneously observed in 38 children (36%), compared with 4 children (9%) in the control group. HBsAg was not eliminated in any of them. The response was worse in patients with a high level of HBV viremia before treatment. However, the initial level of ALT activity did not influence the final outcome of the therapy. The effectiveness of lamivudine was also unaffected by other tested factors, such as: route of transmission, duration of infection, undergone IFN therapy. Lamivudine seems to be an effective and well-tolerated antiviral drug in children with CHB. However, further studies are necessary to optimize this therapy. PMID- 15799861 TI - Treatment of accidental intakes of plutonium and americium: guidance notes. AB - The scientific basis for the treatment of the contamination of the human body by plutonium, americium and other actinides is reviewed. Guidance Notes are presented for the assistance of physicians and others who may be called upon to treat workers or members of the public who may become contaminated internally with inhaled plutonium nitrate, plutonium tributyl phosphate, americium nitrate or americium oxide. PMID- 15799862 TI - Biokinetics of inhaled radioactive methane in rats: a pilot study. AB - Current dosimetric models for radioactive methane assume 1% of inhaled methane is absorbed, all 3H activity is converted immediately to [3H]H2O, and 14C activity is converted immediately to [14C]CO2 (50%) and organically bound carbon (50%). Results of a pilot study using rats to test these models suggest the models overestimate uptake but underestimate organic fixation of 3H and 14C, especially in liver. Also, the biokinetic properties of organically bound 3H and 14C in liver were markedly different from other tissues. Preliminary dose estimates based on observed uptake and organic fixation of label suggest current methane models likely overestimate radiation doses from radioactive methane by 3- to 10 fold. PMID- 15799863 TI - Absorbed dose estimates at the cellular level for 131I. AB - Microdosimetric calculations of 131I have been evaluated for a single cell and for cell clusters. A VsBasic program has been used to calculate stopping power, linear energy transfer, range values and deposited energies per decay for beta particles, Auger and conversion electrons of 131I. The chemical composition of the cell has been taken into account in this model; results were compared with water medium. Besides, total absorbed doses have been calculated for the radionuclides distributed randomly within the cell and clusters. Cross-fire irradiation has been considered for clusters of cells. In this case, absorbed doses per cell within a cluster were found to be significantly higher than absorbed doses per single cell, depending on the cluster size. Results showed that 131I is a promising radionuclide for therapy of tumors from millimeter to centimeter dimensions. PMID- 15799864 TI - Rapid analysis of 14C and 3H in graphite and concrete for decommissioning of nuclear reactor. AB - A rapid oxidizing combustion method using a commercial Sample Oxidizer has been investigated to determine separately the 14C and 3H activities in graphite and concrete. By this method the sample preparation time can be reduced to 2-3 min. The detection limits for 3H and 14C are 0.96 and 0.58 Bq/g graphite and 0.11 and 0.06 Bq/g concrete, respectively. The cross contamination of 14C and tritium in the preparation of samples is less than 0.2%. The interference of other radionuclides in the determination of 14C and tritium in graphite is insignificant. The analytical accuracy, investigated by the standard addition method, is better than 95%. In addition, an acid digestion method has also been used to examine the graphite and concrete activities, to allow comparison with the method developed herein. The two methods show good agreement for graphite samples. Graphite samples were collected from the Danish Reactors DR-2 and DR-3, in addition to two concrete cores drilled in the Danish reactor DR-2; these were analysed for 3H and 14C using the method that has been developed. PMID- 15799865 TI - Scintigraphic imaging with 99mTc- exorphin C in rabbits. AB - Exorphin C is a peptide with five amino acids [(Tyr-Pro-Ile-Ser-Leu) Trifluoroacetate salt] (Sigma) that has an affinity to opioid receptor-expressing tissues and tumors. Exorphin-C was labeled with 99mTc using glucoheptonate (GH) as bifunctional chelating agent. Then, we investigated its radiopharmaceutical potential as opioid receptor-expressing tissue on rabbits. Quality controls were performed by ITLC, paper electrophoresis and HPLC. Labeling efficiency was higher than 98%. The compound was stable for at least 5 h at room temperature. Scintigraphic imaging with 99mTc-GH-exorphin C (99mTc-GE) was performed on male Albino rabbits. Static images were obtained from anterior projection using a Camstar XR/T gamma camera at several time intervals. Although a significant amount of activity was seen in the brain, less activity was seen on receptor saturation studies at 30 min. Slight hepatobiliary excretion was seen, though the main excretion route was renal. After saturating, the receptor hepatobiliary excretion was not seen; the only excretion route was renal. PMID- 15799866 TI - Factors affecting quality control of [18F]FDG injection: bacterial endotoxins test, aluminum ions test and HPLC analysis for FDG and CIDG. AB - High concentrations of citrates and phosphates which are often used in the manufacturing of [18F] fluro-D-glucose (FDG) preparations and wide deviation in the pH value from the neutral level often disturb the detection of endotoxins and aluminum ions using the turbidimetric and aluminum ion paper test method. The column temperature was found to be a major factor influencing the sensitivity of ClDG detection with the HPAEC/PAD method. PMID- 15799867 TI - Preparation of the hypoxia imaging PET tracer [18F]FAZA: reaction parameters and automation. AB - 18F-labeling of the nitroimidazole nucleoside analogue 1-(5-fluoro-5-deoxy-alpha D-arabinofuranosyl)-2-nitroimidazole (FAZA) was developed to use this tracer in PET for detection of hypoxia. Parameters for labeling and hydrolysis were optimized with regard to amount of precursor, temperature and time. Labeling yields reached a maximum of 62+/-4% at 100 degrees C within 5 min using 5 mg of precursor. Hydrolysis was best performed with 1 mL of 0.1 N NaOH at 20 degrees C for 2 min. Transfer of these conditions to an automated synthesizer resulted in an overall radiochemical yield of 20.7+/-3.5%. Absolute yields at EOS were 9.8+/ 2.3 GBq of [18F]FAZA ready for injection (n=21; 50 min after EOB; irradiation parameters: 35 microA, 60 min). Thus, a convenient approach suitable for large scale production of [18F]FAZA was developed by an automated process. PMID- 15799868 TI - Synthesis, radiosynthesis, in vitro and preliminary in vivo evaluation of biphenyl carboxylic and hydroxamic matrix metalloproteinase (MMP) inhibitors as potential tumor imaging agents. AB - Excess matrix degradation is one of the hallmarks of cancer and is an important factor in the process of tumor progression. It is implicated in invasion, metastasis, growth, angiogenesis and migration. Many characteristics of matrix metalloproteinases (MMPs) make them attractive therapeutic and diagnostic targets. MMP expression is upregulated at the tumor site, with localization of activity in the tumor or the surrounding stroma, providing a target for medical imaging techniques. Radioiodinated carboxylic and hydroxamic MMP inhibitors 2-(4' [123I] iodo-biphenyl-4-sulfonylamino)-3-methyl-butyric acid (9) and 2-(4'-[123I] iodo-biphenyl-4-sulfonylamino)-3-methyl-butyramide (11), their unlabelled standards and precursors were synthesized. Radioiodination was conducted by electrophilic aromatic substitution of the tributylstannyl precursors and resulted in radiochemical yields of 70+/-5% (n=6) and 60+/-5% (n=4), respectively. In vitro zymography and enzyme assays showed for both hydroxamic acid and carboxylic acid compounds a good inhibition activity and a high selectivity for MMP-2. In vivo biodistribution in NMRI mice showed no long-term accumulation in organs and the possibility to accumulate in the tumor in a later phase of this study. PMID- 15799869 TI - A kit for labeling of [188Re] human serum albumin microspheres for therapeutic use in nuclear medicine. AB - Intra-arterial infusion of labeled particles is an effective method for local endoradiotherapy of tumors. In this study, we present an efficient method of radiolabeling biodegradable human serum albumin (HSA) microspheres with the short lived beta-emitter 188Re using a simple and reliable kit. Up to now, it was not possible to label particles with more than 85-90% efficiency. Under optimized reaction conditions, we achieved nearly 100% uptake of 188Re. Using the proposed kit, we added tartrate solution and increased the pH from 2 to 4-5 after 1 h reaction time at 95 degrees C. The 188Re binding to the particles was found to be stable in vitro. PMID- 15799870 TI - Facile synthesis of bis(hydroxamamide)-based tetradentate ligands for 99mTc radiopharmaceutical. AB - A facile, two-step synthesis of the bis(hydroxamamide)-based tetradentate ligands for 99mTc-radiopharmaceuticals is described. Firstly, the hydroxamamide was converted to hydroximic acid chloride by reaction with sodium nitrite in hydrochloric acid at 0 degrees C. Secondly, treating the halide with the ethylenediamine or 1,3-propylenediamine in absolute ethanol formed the desired products, N,N'-ethylene bis(1-(4-nitroimidazole-1-yl)-propan-hydroxyiminoamide) (I) and N,N'-propylene bis(1-(4-nitroimidazole-1-yl)-propanhydroxyiminoamide) (II). The corresponding 99mTc complexes showed high yields and were found by paper electrophoresis to be electrically neutral under physiological conditions. The partition coefficients indicated a distinct difference between the two complexes. PMID- 15799871 TI - In situ measurements of the sub-surface gamma dose from Chernobyl fallout. AB - Methods of estimating external radiation exposure of soil-dwelling organisms are currently of much research and regulatory interest. In this paper, we report the first in situ measurements of the sub-surface gamma dose rate for 137Cs contaminated land that quantify variation in dose rate with depth. Two contrasting sites have been investigated. The first site comprised a mineral type soil with a low percentage of organic matter and the second site chosen was in a peat-bog. The different soil compositions afford different 137Cs mobility and this results in variations in the measured gamma dose-rate with soil depth. For each site the paper reports the measured dose rates, the 137Cs activity depth profile, the 137Cs inventory and a description of the soil-characteristics. It is suggested that these data can be used to produce estimates of the sub-surface gamma dose rate in other sites of 137Cs contamination. PMID- 15799872 TI - Statistical evaluation of FRAM gamma-ray isotopic analysis data. AB - High-purity germanium (HPGe) detector gamma-ray spectra were analyzed using the FRAM (fixed energy, response function analysis with multiple efficiencies) gamma ray isotopic analysis software. The analyses are based on multiple measurements of samples having well-documented isotopic composition from mass spectrometry measurements. Statistical analyses of the FRAM results are reported, the errors in FRAM analyses arising from the choice of detector type and the energy region are discussed, and the errors that resulted from sample-dependent and analysis dependent effects are quantified. PMID- 15799873 TI - Towards models for analysis of time-resolved luminescence spectra from quartz. AB - Simple and stretched-exponential models for use in the analysis of time-resolved luminescence spectra from quartz are compared. The methods have been applied to evaluate lifetimes from portions of time-resolved luminescence spectra measured either during or after the luminescence stimulating pulse. We discuss the basis for use of the exponential functions as well as examine the application of, in this context, atypical stretched-exponential functions for analysis of time resolved luminescence spectra from quartz. PMID- 15799874 TI - Direct evidence of the temperature dependence of Gd-BOPTA transport in the intact rat liver. AB - The aim was to study the influence of temperature on the transport of the hepatobiliary contrast agent Gadobenate dimeglumine (Gd-BOPTA). Rat livers were isolated and perfused with Gd-BOPTA at 12, 25, 30, 36 and 38 degrees C. After the perfusion period, biopsies were collected and the MR signal intensity was measured. Uptake and biliary excretion were quantified with radiolabeled Gd BOPTA. MR signal intensity decreased with temperature of perfusion. This phenomenon was appropriately quantified with 153Gd and 153Sm labeling, in contrast to 67Ga. PMID- 15799875 TI - Effects of high-frequency electromagnetic fields emitted from card readers of access control systems on electronic pocket dosimeters. AB - High-frequency electromagnetic fields in the 120 kHz band emitted from card readers for access control systems caused abnormally high doses on electronic pocket dosimeters (EPDs). All EPDs recovered their normal performance by resetting after the exposure ceased. The electric and magnetic immunity levels of the EPDs were estimated by using the distances needed to prevent electromagnetic interference. PMID- 15799876 TI - Comparison of outdoor activity size distributions of 220Rn and 222Rn progeny. AB - Inhalation of 222Rn and 220Rn progeny from the domestic environment contributes the greatest fraction of the natural radiation exposure to the public. Dosimetric models are most often used in the assessment of human lung doses due to inhaled radioactivity because of the difficulty in making direct measurements. These models require information about the parameters of activity size distributions of thoron and radon progeny. The present study presents measured data on the attached and unattached activity size distributions of thoron and radon progeny in outdoor air in El-Minia, Egypt. The attached fraction was collected using a low-pressure Berner cascade impactor technique. A screen diffusion battery was used for collecting the unattached fraction. Most of the attached activities for 222Rn and 220Rn progeny were associated with aerosol particles of the accumulation mode. The activity size distribution of thoron progeny was found to be shifted to slightly smaller particle size compared to radon progeny. PMID- 15799877 TI - A solid-state NMR application of the anomeric effect in carbohydrates: galactosamine, glucosamine, and N-acetyl-glucosamine. AB - Simple 2D 13C/15N heteronuclear correlation solid-state NMR spectroscopy was implemented to resolve the 15N resonances of the alpha and beta anomers of three amino monosaccharides: galactosamine (GalN), glucosamine hydrochloride (GlcN), and N-acetyl-glucosamine (GlcNAc) labeled specifically with 13C1/15N spin pairs. Although the 15N resonances could not be distinguished in normal 1D spectra, they were well resolved in 2D double CP/MAS correlation spectra by taking advantage of the 13C spectral resolution. The alpha and beta resonances shifted apart by 3-5 ppm in their 13C chemical shifts, and differed by 1-2 ppm in the extended 15N dimension. Aside from this, the detection of other 13C/15N correlations over short distances was also achieved arising from the C2, C3 and CO carbons present in natural abundance. 2D double CP/MAS chemical shift correlation NMR spectroscopy is a simple and powerful technique to characterize the anomeric effect of amino monosaccharides. Applications of the 2D method reveal well resolved 15N and 13C chemical shifts might be useful for structural determination on carbohydrates of biological significance, such as glycopeptide or glycolipids. PMID- 15799878 TI - Correlation between 1H FID and T1rho components in heterogeneous polymer systems: an application to SBS. AB - Wideline 1H FID and relaxation measurements of a relatively simple motionally heterogeneous system, the triblock copolymer styrene-butadiene-styrene, have been performed in a temperature range between the polystyrene and polybutadiene glass transition temperatures. The two FID and the two spin lattice relaxation time in the rotating frame (T1rho) components found at each temperature have been correlated by means of a two-dimensional approach. It is shown that this approach allows dynamic information, not accessible simply by interpreting proton T1 and T1rho data, to be revealed. In the case examined, the correlation found could be confirmed by high-resolution 1H T1rho-selective 13C Cross Polarization experiments. PMID- 15799879 TI - High-resolution solid-state 119Sn and 195Pt NMR studies of MPtSn semiconductors (M = Ti, Zr, Hf, Th). AB - Solid-state 119Sn and 195Pt magic-angle spinning (MAS) NMR spectra are reported on a series of MPtSn compounds (M = Ti, Zr, Hf, Th). In favorable cases (TiPtSn and ZrPtSn) the spectra reveal expected J-coupling patterns originating from indirect spin coupling between Pt and Sn nuclei. MAS has no effect on the broad and asymmetric spectra of either 119Sn and 195Pt nuclei in HfPtSn. PMID- 15799880 TI - Measurement of J-couplings between spin-(1/2) and quadrupolar nuclei by frequency selective solid-state NMR. AB - We report a REDOR-based scheme for the measurement of heteronuclear J-couplings in solid samples with well defined structure, containing spin-12 and quadrupolar nuclei, which can be used with selective RF irradiation to target a specific spin pair, and which provides direct information about the number of coupled spins. PMID- 15799881 TI - Calculation of bridging oxygen 17O quadrupolar coupling parameters in alkali silicates: a combined ab initio investigation. AB - Ab initio band-structure calculations based on the density functional theory have been performed for several crystalline Li, Na, and K-silicates to obtain electric field gradients (efg) for oxygen atoms. The efg for bridging oxygen environments in these compounds were also investigated by performing ab initio self-consistent field Hartree-Fock molecular orbital calculations on silicate clusters, and there is good agreement between these two approaches. By performing additional ab initio quantum chemistry calculations on model silicate clusters the factors influencing the 17O quadrupole coupling parameters for bridging oxygen environments in alkali silicates have been examined. The quadrupolar asymmetry parameter was found to be dependent on the Si-O-Si angle and the nature of the modifier cation, in agreement with previous studies. In contrast, the quadrupolar coupling constant was found to have a strong dependence on Si-O distance, as well as Si-O-Si angle and the nature of the modifier cation. Analytical expressions describing these dependencies are proposed, which should assist in describing the local environments of bridging oxygen in crystalline and amorphous materials. PMID- 15799882 TI - Studies of silicophosphate derivatives by 31P-->29Si CP MAS NMR. AB - We show that it is possible to efficiently transfer magnetization from 31P to 29Si, using variable amplitude CP MAS experiment. This experiment is demonstrated by using Si5O(PO4)6, the synthesis protocol of which is described. From the obtained results, we show that the experiment allows the spectral edition of 29Si spectra from 31P-->29Si CP, enabling the studies of derivatives involving Si-O-P linkages, such as phosphosilicate glasses, microporous silicoaluminophosphates (SAPO) and bioactive phosphosilicates. PMID- 15799883 TI - A study of the headgroup motion of sphingomyelin using 31P NMR and an analytically soluble model. AB - A 31P NMR investigation has been carried out of the headgroup dynamics of sphingomyelin molecules in bilayers for the L alpha and L beta' phases. The resulting line shapes have been analysed in terms of a reduced-parameter model, using van Faassen's method for obtaining an analytic solution to first-order stochastic differential equations to simulate the line shapes of oriented and non oriented samples. Our treatment results in good fits to measured data but using fewer parameters than traditional methods. Angles and correlation times (tau parallel and tau perpendicular) describing the geometry and dynamics of the headgroup are obtained by optimising the agreement between simulated and experimental data. The results are contrasted with those obtained for the lecithins DMPC and DPPC using a similar analysis. Not only are tau parallel and tau perpendicular now equal in value for the L alpha phase, but this value is also found to be nearly four times larger than the longest correlation time for the lecithins. We interpret this as evidence of inter- and intramolecular hydrogen bonding in the L(alpha) phase of sphingomyelin. In the L beta' phase of sphingomyelin, however, although tau(parallel) and tau(perpendicular) remain equal their value is now 32% smaller than that of the lecithins in the same phase. This indicates less difference between the fluidities in the headgroup region of the two phases of sphingomyelin as compared to that of the lecithins. Another significant difference between the L beta' phases is that the tilt angle for sphingomyelin is found to be nearly twice as large as for the lecithins. We argue that these combined observations point to the existence of hydrogen bonding in this phase also. Again in contrast to our previous work on lecithins, our results provide evidence of a negative diamagnetic anisotropy in sphingomyelin molecules, even in the L beta' phase. This is provided for in our model by the assumption that our unoriented samples consist of prolate ellipsoidal liposomes whose major axes are oriented parallel to the static magnetic field. The apparently different diamagnetic behaviour of sphingomyelin in the present work is due to the higher static field used rather than any intrinsic difference in this respect between sphingomyelin and the lecithins DMPC and DPPC. PMID- 15799884 TI - Orientational deconvolution of two-dimensional static disorder by a Tikhonov regularized method for 2H solid state NMR of nano-tubular-oriented structures. AB - A general trend of supramolecular chemistry is the building of mesoscopic oriented nanotubes. When the typical radius of the channel almost fits the mean supramolecular cross-sectional radius, the guest supramolecule is ultraconfined in the channel. In that case, only rotational disorder around the channel axis is allowed. For sufficiently low temperatures, we expect this 2D disorder to be static on the NMR timescale, and it should reflect the local symmetry of the channels. In this article, we show that experiments performed with deuterium single crystal solid-state NMR as a function of rotation around the channel axis may lead to important information concerning the static orientational disorder of the confined supramolecules. Using an inversion method with Tikhonov regularization and a positivity constraint, that also takes into account of the nonideal pulse sequence response, the orientational probability density can be obtained from 1D experiments performed at different angles, even for significant disorder. As a first step, the method is validated on four different theoretical distributions. It is then applied to 2H NMR single crystal experiments performed on an archetype of parallel channels intergrowth compounds: selectively deuterated 1,10-decanedicarboxylic diacids in hydrogenated urea channels. The diacids form one-dimensional infinite hydrogen-bonded chains ultraconfined in urea linear channels. PMID- 15799885 TI - Double-quantum NMR spectroscopy based on finite pulse RFDR. AB - We demonstrate that the finite pulse RFDR sequence (J. Chem. Phys. 114 (2001) 8473) can be used effectively for 31P double-quantum NMR spectroscopy at a spinning frequency of 10 kHz. The 31P NMR data measured for hydroxyapatite and octacalcium phosphate show that sizable double-quantum excitation efficiency can be obtained with the ratio of the recoupling field to spinning frequency set equal to 1.67. PMID- 15799886 TI - Have PhD, will travel. AB - This autobiography contains some of the clinical and research highlights of my career in the context of my personal odyssey and my appreciation of science as a potential check on political and economic forces in psychology. Typically, my research arose from clinical problems encountered at work and from my disinclination to rely on methods that lacked prior demonstrated validity for the type of problem and client in question. Otherwise, I feared my recommendations were no better than those based on common prejudices and biases, which I encountered throughout my work life as a dark-skinned, outspoken, non-Christian from the Bronx. PMID- 15799887 TI - An illustration of multilevel factor analysis. AB - The factor analysis of repeated measures psychiatric data presents interesting challenges for researchers in terms of identifying the latent structure of an assessment instrument. Specifically, repeated measures contain both within and between individual sources of variance. Although a number of techniques exist for separating out these 2 sources of variance, all are problematic. Recently, researchers have proposed that exploratory multilevel factor analysis (MFA) be used to appropriately analyze the latent structure of repeated measures data. The chief objective of this report is to provide a didactic step-by-step guide on how MFA may be applied to psychiatric data. In the discussion, we describe difficulties associated with MFA and consider challenges in factor analyzing life event appraisals in psychiatric samples. PMID- 15799888 TI - The interpersonal circumplex as a model of interpersonal capabilities. AB - In this study, we sought to challenge the existing conceptualization of interpersonal capabilities as a distinct construct from interpersonal traits by explicitly taking into account the general factor inherent within most models of circumplexes. A sample of 206 college students completed a battery of measures including the Battery of Interpersonal Capabilities (BIC; Paulhus & Martin, 1987). Principal components analysis and the randomization test of hypothesized order relations demonstrated that contrary to previous findings, the BIC adhered to a circular ordering. Joint analysis of the BIC with the Interpersonal Adjective Scale (Wiggins, 1995) using principal components analysis and structural equation modeling demonstrated that the 2 measures represented similar constructs. Furthermore, the general factor in the BIC was not correlated with measures of general self-competence, satisfaction with life, or general pathology. PMID- 15799889 TI - Stability in optimism-pessimism in relation to bad news: a study of women with breast cancer. AB - In this study, we investigated the stability of optimism-pessimism in relation to receiving bad news (i.e., positive lymph nodes, more advanced cancer stage) after breast cancer surgery and in comparison to the stability of anxiety and depression level. The women (n = 165) completed the Life Orientation Test-Revised (Scheier, Carver, & Bridges, 1994) and the Hospital Anxiety and Depression Scale (Zigmond & Snaith, 1983) at time of diagnosis, 3 months, and 12 months after surgery. Overall, women's optimism-pessimism levels remained stable over the follow-up period whether they received bad news or not. In contrast, women's anxiety and depression level decreased significantly over the same time period with 1 exception. Women's level of depression remained fairly stable among those who received bad news. Optimism-pessimism appears to remain stable over situations as well as time and regardless of negative affects. PMID- 15799890 TI - Assessment of combat-related stress and physical symptoms of Gulf War veterans: criterion validity of selected hand test variables. AB - We examined the utility of selected Hand Test (Wagner, 1983) variables in relation to posttraumatic stress and physical symptoms in Gulf War (GW) veterans. In this study, we sought to replicate and expand on prior empirical findings that have demonstrated efficacy of the Hand Test in the assessment of posttraumatic stress disorder (PTSD; Walter, Hilsenroth, Arsenault, Sloan, & Harvill, 1998). Based on this previous research, Hand Test variables were selected a priori and examined across three groups of veterans: (a) a control group of participants who were in a reserve unit not deployed to the GW theater of operations, (b) a subclinical group of deployed GW veterans who reported 1 to 5 Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) criteria for PTSD, and (c) a group of deployed GW veterans who met DSM-IV criteria for PTSD. Analyses demonstrated significant differences across the three groups and significant relationships among selected Hand Test variables with the number of DSM-IV symptoms of PTSD reported in the interviews as well as with the number of physical problems reported by these veterans. We discuss these findings in relation to the assessment and treatment of posttraumatic stress symptomatology. PMID- 15799891 TI - Symptom correlates of MMPI-2 scales and code types in a private-practice setting. AB - The purpose of this study was to determine empirical symptom correlates of MMPI-2 (Butcher et al., 2001) scales for private-practice clients, a very understudied yet important population. We examined the scores of 240 male and 407 female clients on the Clinical scales, Content scales, Supplementary scales, and frequently occurring code types. We used a factor analyzed version of the Adult Clinical scales of the Multiaxial Diagnostic Inventory (Doverspike, 1990) as a criterion measure. The results generally indicated that the symptom correlates for the MMPI-2 scales and code types were quite similar to those that have been previously reported in other outpatient and inpatient settings. We concluded that descriptors of MMPI-2 scales and code types generated in other settings may be used when interpreting MMPI-2 profiles of private-practice clients. PMID- 15799892 TI - Validity of a short form of the Achievement-Motive Grid (AMG-S): evidence for the three-factor structure emphasizing active and passive forms of fear of failure. AB - My primary objective in this research was to examine diverse validity aspects of a short form of the Achievement-Motive Grid (AMG-S; Schmalt, 1999). The AMG is a picture-based measure that combines features of the Thematic Apperception Test (TAT; Murray, 1943) with features of self-report questionnaires. Similar to the TAT, the motive gets aroused by presenting a set of pictured achievement situations. Similar to a questionnaire, the motive is measured with a set of statements representing characteristic components of the achievement motive. Quite in line with the original version, factor analyses of the AMG-S all recommend the adoption of a 3-factor solution in which 1 hope-of-success (HS) and 2 fear-of-failure (FF) concepts are differentiated. The HS and passive FF factor (FFp) are based on positive and negative efficacy evaluations, respectively. The active FF factor (FFa) describes anticipations of impending failure. FFp is clearly a "withdrawal" factor, whereas FFa is clearly an "invigoration" factor. In the construct validity section of this research, I focus on classical and recent elaborations of approach and avoidance motivation in which I show that HS, FFp, and FFa play a substantial role in a motive-goals-outcomes framework of achievement motivation. PMID- 15799893 TI - Psychometric properties of the Revised Cheek and Buss Shyness Scale. AB - Although the Revised Cheek and Buss Shyness Scale (RCBS; Cheek, 1983) is widely used, its psychometric properties largely are unknown. In this investigation, we examined the normative data, factor structure, internal consistency, test-retest reliability, and convergent/discriminant validity of the RCBS using a sample of 261 university students. Results provided strong support for the stability of normative data over time, reliability of the measure, and its predicted associations with contemporary measures of shyness, social anxiety, and related constructs. Although support was obtained for a unifactorial conceptualization of shyness, an exploratory factor analysis revealed an alternative 3-factor solution that was supportive of a previously proposed meta-analytic model of shyness (Jones, Briggs, & Smith, 1986) and was consistent with other prominent shyness theories (Buss, 1980; Pilkonis, 1977a, 1977b; Zimbardo, 1977). This factor model was replicable on a holdout sample, and there were some data to support the discriminant validity of factors. PMID- 15799894 TI - Linking personality disorders and clinical syndromes on the MCMI-III. AB - We examined the relationship between personality disorders (PDs) and clinical syndromes (CSs) as measured by the Millon Clinical Multiaxial Inventory-III (MCMI III; Millon, 1997) in a large, heterogeneous sample of psychiatric patients (N = 2,366) who completed the instrument as part of routine assessment following presentation for treatment. Using separate sets of base rate (BR) and nonoverlapping scale scores, we factor analyzed the PD and CS scales together and then separately. We correlated results from the latter analyses to determine how trait dimensions were associated with syndrome dimensions. We also studied co occurrence at the scale level by examining CS score profiles of patients who were grouped according to their highest PD scale elevation > or = BR75. Results for the two score sets were very similar and were consistent with previous research on the MCMI-III and its predecessors that identified 3 underlying dimensions loading both PD and CS scales. Three fourths (76.2%) of the sample had a highest PD scale > or = BR75, and among these, 90% had at least 1 CS scale > or = BR75, whereas 62.4% had 3 or more CS scales above this elevation. Findings underscore the substantial overlap between PDs and CSs along 3 dimensions that resemble Horney's (1945) tripartite interpersonal distinction of moving toward, away, and against, as well as Eysenck's (1994) higher order factors of neuroticism, extraversion, and psychoticism. PMID- 15799895 TI - Supervising personality assessment: the integration of intersubjective and psychodynamic elements in the supervisory process. AB - The supervision of personality assessment can be likened to the process of teaching art. Much like artistic training, supervision of personality assessment is a complex process that calls for creative integration of different working processes, some more technical and formal and others broader and more complex. On the surface, compared to psychotherapy supervision, supervision of personality assessment is fairly structured and centered on specific tasks such as scoring, making a diagnosis, or determining personality organization. Nevertheless, principles of psychodynamic psychotherapy such as parallel processes are relevant as well. We discuss the process of integrating intersubjective and psychodynamic elements into personality assessment supervision and argue that this integration helps the supervisory work to proceed effectively. We present the role of knowledge, the institution, and parallel processes in the supervisory work. PMID- 15799897 TI - Brain magnetic resonance imaging abnormalities following off-pump cardiac surgery. AB - BACKGROUND: Neurological and cognitive deficits are known complications after coronary artery bypass surgery (CABG) and are believed to be secondary to brain ischemia. Diffusion-weighted magnetic resonance imaging (DW-MRI) of the brain is especially sensitive and can depict ischemic areas that may not be evident clinically or with conventional MRI. Abnormalities found at brain MRI following CABG performed with cardiopulmonary bypass (ie, on pump) have been reported, but data are limited for CABG performed without use of cardiopulmonary bypass (ie, off pump). The objective of this study was to determine the rate of DW-MRI detection of brain lesions following off-pump CABG. METHODS AND RESULTS: Sixteen patients consecutively undergoing off-pump CABG underwent DW-MRI prior to and after surgery. A neuroradiologist blinded to patient data coded the location and size of lesions. Neurological function was assessed with the National Institutes of Health Stroke Scale. Five (31%) of the patients had new focal ischemic lesions found at postoperative DW-MRI. Risk factors for postoperative brain ischemic lesions were similar between patients with and those without lesions found at DW MRI. No patient had a definite clinical stroke after surgery. CONCLUSIONS: Ischemic lesions found at DW-MRI are seen after off-pump CABG at a rate similar to that reported for CABG with cardiopulmonary bypass. This finding suggests that these lesions are not totally due to cardiopulmonary bypass. Further prospective clinical studies would be helpful for determining which factors are causally related to brain ischemia following CABG. PMID- 15799898 TI - Noncompaction of ventricular myocardium in a patient with congenitally corrected transposition of the great arteries treated surgically: case report. AB - Noncompaction of the ventricular myocardium is a rare disorder that represents numerous prominent trabeculations and intratrabecular recesses in the ventricles. It is believed to represent not only an arrest in endomyocardial morphogenesis but also an unclassified cardiomyopathy. The pathology has been almost invariably associated with other congenital cardiac malformations. A female patient with noncompaction of the myocardium of both ventricles and congenitally corrected transposition of the great arteries (cTGA), situs inversus totalis, and atrial and ventricular septal defects is described. When she was 7 days old a permanent pacemaker was implanted because of complete heart block. Prazosin (Minipress), an alpha-receptor blocker, was administered, and the cardiac ejection fraction showed a striking increase from 20% to 42%. Despite careful and regular follow-up evaluations, the general condition of the patient slowly worsened. Five months after surgery she died of hepatorenal failure and low cardiac output. This case report is thought to be the first description of congenital complete heart block, cTGA, and situs inversus totalis with noncompaction of the myocardium of both ventricles. PMID- 15799899 TI - An unusual case of thyroid Hurtle cell carcinoma with direct extension to the right brachiocephalic vein, right auricle, and right atrium: case report. AB - The clinical behavior of various types of thyroid tumor have been much studied during the past several decades, and the histologic features, surgical management, and prognostic factors of follicular and papillary tumors in particular have been clarified to a considerable degree. On the other hand, there is still controversy concerning management of Hurtle cell tumor (HCT) of the thyroid. HCT is not a common disease, making it impossible to obtain sufficient clinical data at a single institution. It has been reported that all HCT are aggressive and should be treated as malignant tumors. It is believed that an accurate differential diagnosis can be made between cancer and adenoma on the basis of pathological studies. We describe a patient with HCT of the thyroid extending into the right atrium. To our knowledge, after a Medline search, this is the first such case in the medical literature. The interesting features are described, and the relevant literature is briefly reviewed. PMID- 15799900 TI - Remission of high-output heart failure after surgical repair of 30-month arteriovenous femoral fistula: case report. AB - We present a 15-year-old male patient who was admitted to our hospital because of breathlessness and palpitations at minimal physical effort (New York Heart Association class II). The patient had a history of an abdominal and left thigh firearm wound that was surgically treated 30 months earlier. Auscultation over the left femoral groin region revealed a systolodiastolic murmur. X-ray examination of the chest demonstrated significant cardiomegaly. Transthoracic echocardiography revealed an enlargement of 4 cardiac chambers, as well as significant mitral and tricuspid regurgitation. Vascular ultrasound of the femoral artery and vein confirmed the diagnosis of a traumatic arteriovenous fistula. The patient underwent surgical correction of the fistula, after which the symptoms subsided rapidly. Follow-up echocardiography performed 2 months after surgical repair showed a substantial reduction of cardiac size and a nearly complete absence of valvular regurgitations. This case highlights the importance of the recognition of arteriovenous fistulas as a cause of unexpected heart failure and demonstrates that the condition may improve substantially and rapidly after fistula correction. PMID- 15799901 TI - The technical aspect of the gastroepiploic artery graft skeletonization with the harmonic scalpel: the samurai technique. AB - A novel skeletonization technique using the scissors-type harmonic scalpel (Ethicon Endo-Surgery, Cincinnati, OH, USA) is presented. This "samurai technique," which uses the harmonic scalpel by frequently turning over the scissors, facilitates the handling of the gastroepiploic artery, enlarges the caliber size, and allows easy skeletonization without any vessel injury. PMID- 15799902 TI - Synergy of old and new technology results in successful revascularization of the anterior myocardium with relief of angina in the absence of suitable targets. AB - BACKGROUND: Diffuse and distal left anterior descending (LAD) coronary disease that is refractory to conventional surgical and/or percutaneous revascularization represents a clinical and economic dilemma. Transmyocardial laser revascularization (TMLR) has improved angina without clear measurable improvement of myocardial perfusion. This study was undertaken to determine if combining a Vineberg implant with TMLR of the LAD distribution enhances myocardial perfusion and relieves symptoms. METHODS: Twenty-one patients with an obliterated LAD and a viable anterior wall underwent off-pump coronary artery bypass grafting (OPCAB) (2.6 grafts/patient). Eight were studied with preoperative, postoperative-early (4-9 days), and postoperative-late (3-5 months) stress and rest nuclear imaging. In all but 3 cases, the Vineberg implant was modified such that the distal end of the conduit, as it emerged from the muscular tunnel, was anastomosed to any patent LAD segment. The anterior wall, to the left of the LAD, was also instrumented with a Holmium yttriumaluminum- garnet laser (8-16 sites). RESULTS: There has been 100% follow-up with durations ranging from 6 to 36 months. There were no deaths. All patients had complete relief of their angina. Serial perfusion scans demonstrated a 2-phase improvement in perfusion. Three of the patients underwent angiography of the implant at 6 to 9 months; angiography in each case demonstrated a patent robust conduit. The 1 patient studied at 24 months demonstrated several sites of a myocardial "blush" consistent with neovascularization. CONCLUSIONS: Although some of the benefits of TMLR/Vineberg may be a consequence of collateral blood flow from other revascularized regions, we believe there to be a synergistic effect on perfusion and angina relief by these combined procedures which may be related to angiogenesis. PMID- 15799903 TI - Initial experience of an automated anastomotic distal device in off-pump CABG. AB - BACKGROUND: Recent progress in minimally invasive technology in the field of coronary artery bypass grafting (CABG) stimulates interest in anastomotic devices used to facilitate distal coronary anastomosis. We assessed the feasibility of the automated anastomotic distal device (AADD) on arterial grafts in patients who underwent off-pump CABG (OPCAB) and evaluated the early anastomotic patency and clinical results of the AADD based on an elliptical nitinol ring with attached 8 pins. METHODS: Fourteen patients scheduled for multivessel OPCAB using arterial grafts between August 2003 and February 2004 were studied. Among 19 patients enrolled, 5 patients were excluded: 2 patients because of failure of graft flaring onto the implant pins, 2 because of small and diseased target coronary artery (<1.5 mm in diameter), and 1 because of conversion to cardiopulmonary bypass. The distal anastomosis using the AADD was performed for the nondominant coronary artery. RESULTS: The total number of distal anastomoses was 48 (34 hand sewn sutures and 14 AADD sutures), and the average number of distal anastomoses per patient was 3.4 +/- 1.0. The grafts used for the AADD were right gastroepiploic artery (RGEA) in 10 patients, saphenous vein anastomosed to the end of the RGEA in 2 patients, and internal thoracic artery in 2 patients. The average time required for distal anastomosis using the AADD (from arteriotomy to anastomosis completion) was 2.9 +/- 0.7 minutes (range, 1.5-4 minutes). The mean flow and pulsatility index of the AADD grafts measured intraoperatively by transit time flow measurement were 20.0 +/- 10.3 mL/min and 2.4 =/- 1.2, respectively. Early postoperative coronary angiographies demonstrated widely patent grafts in 32 of 34 hand-sewn anastomoses and 13 of 14 AADD sutures. There were no adverse events related to the use of the device. CONCLUSIONS: Our initial experience demonstrated that distal anastomosis using the AADD was feasible in most of the patients who underwent OPCAB using arterial grafts. Distal anastomosis using the AADD had the advantage of shortening the actual suturing duration and might provide a method for standardizing the anastomotic procedure. PMID- 15799904 TI - A rare presentation of cardiac hydatid cyst: stroke and acute aortic occlusion. AB - Cardiac involvement in hydatid disease is uncommon. We report a case of a surgically treated ruptured left ventricular hydatid cyst, which presented with acute stroke and was later complicated by distal aortic embolism due to perioperative dislodgement of the germinative membrane. PMID- 15799905 TI - Surgery using median sternotomy for diffuse aneurysmal disease of the thoracic aorta. AB - We present our experience using an anterior approach for the replacement of an extensive aneurysm of the thoracic aorta. In recent years we have performed surgery on 20 patients by means of a median sternotomy for aneurysms of the ascending aorta, aortic arch, or descending thoracic aorta. In all but 1 of the patients, a procedure in the ascending aorta was also performed. In some patients a small anterior left thoracotomy at the 4th intercostal space was required to allow the replacement of the aorta as for the diaphragm. Antegrade selective cerebral perfusion (ASCP) according to Kazui's technique was used as a brain protection method. All procedures were performed successfully and the aneurysm was completely resected. No neurologic complications or other major complications occurred. We believe that the anterior approach for extensive thoracic aorta replacement is feasible, allows the use of ASCP, and has shown encouraging results. PMID- 15799906 TI - Advantages of off-pump coronary artery bypass grafting in long-term hemodialysis patients: multicenter analysis. AB - OBJECTIVE: Long-term hemodialysis remains a major risk factor for coronary artery bypass grafting (CABG). Off-pump CABG (OPCAB) is expected to offer benefits for these high risk patients; however, this issue has not been clarified. To elucidate the issue, we conducted a multicenter retrospective review of long-term hemodialysis patients who underwent on- or off-pump CABG. PATIENTS: Between January 1998 and December 2002, 53 hemodialysis patients underwent elective CABG at 14 centers. Nineteen patients underwent OPCAB, and 34 patients underwent conventional CABG (CCAB). Preoperative and perioperative variables, morbidity, and mortality were compared in the 2 groups. There were no significant differences in preoperative variables between the 2 groups. RESULTS: The length of intensive care unit (ICU) stay (3.7 versus 5.9 days), the amount of blood loss (668 versus 1100 mL), and the amount of red cell transfusion (4.7 versus 12.2 units) were less in the OPCAB group. The perioperative morbidity was significantly lower in the OPCAB group (0.0% versus 26.5%). The hospital mortality was not significant but was low in the OPCAB group (0% versus 14.7%, P = .079). CONCLUSION: OPCAB significantly decreased blood loss, blood transfusion, ICU stay, and major perioperative morbidity compared with the values for CCAB. OPCAB may have advantages over CCAB in long-term hemodialysis patients. PMID- 15799907 TI - A novel device for clampless proximal anastomosis in OPCAB surgery: the IPAD. AB - The Coalescent IPAD (Coalescent Surgical, Sunnyvale, CA, USA) is a novel device that enables creation of a compliant proximal vein anastomosis to the aorta during off-pump coronary artery bypass (OPCAB) surgery while reducing the risk of cerebrovascular emboli and early stenosis. We performed IPAD-created anastomoses in 76 patients having standard OPCAB surgery. Following the procedures, no patients developed myocardial infarctions or reported angina, suggesting patency of the vein grafts to be of high quality. We found the device to be a useful adjunct for minimally invasive CAB graft surgery. PMID- 15799908 TI - Optimizing intraoperative cerebral oxygen delivery using noninvasive cerebral oximetry decreases the incidence of stroke for cardiac surgical patients. AB - BACKGROUND: A recent study demonstrated that almost 75% of strokes after coronary artery revascularization surgery occur in patients classified preoperatively as low to medium risk. Thus, despite the use of risk classification, most strokes can occur when not expected. We hypothesized that optimization of cerebral oxygen delivery variables by using noninvasive cerebral oximetry could reduce the incidence of stroke. METHODS: Cerebral oximetry was used by all surgeons to monitor cerebral oxygen saturation in all cardiac surgery patients from January 1, 2002, until June 30, 2003 (n = 1034; 18 months, treatment group). Cerebral oxygen delivery was optimized during surgery by modifying oxygen delivery and consumption variables to maintain oximetry values at or near the patient's preinduction baseline. Stroke was defined according to guidelines of the Society of Thoracic Surgeons. The incidence of stroke in the treatment group was compared with that for patients who underwent cardiac surgery between July 1, 2000, and December 31, 2001, (n = 1245; 18 months, control group) before cerebral oximetry was incorporated. RESULTS: Age and sex distribution were similar in the 2 groups. The study group had significantly more patients in New York Heart Association (NYHA) classes III and IV than the control group, and patients in the study group were sicker overall. Despite this difference, the study group overall had fewer permanent strokes (10 [0.97%] versus 25 [2.5%]; P < .044). This difference remained significant when the results were controlled for NYHA class and on-pump or off-pump surgery. When the patients were examined by NYHA class, the proportion of patients requiring prolonged ventilation was significantly smaller in the study group (6.8% versus 10.6%; P < .0014), as was the length of hospital stay (P < .046). CONCLUSIONS: The treatment group, which underwent all cardiac surgeries with optimized cerebral oxygen delivery using cerebral oximetry monitoring, demonstrated a significantly lower incidence of permanent stroke. Because our study is retrospective, a prospective randomized trial is warranted. PMID- 15799909 TI - Total arterial revascularization with a single y-composite graft for triple vessel disease: comparison of 2- and 3-artery grafts. AB - BACKGROUND: Treating triple-vessel disease by grafting a single Y-composite graft with 1 arterial conduit attached to the side of the internal thoracic artery is sometimes not used because of the possibility of hypoperfusion. METHODS: Between March 2001 and June 2003, 271 patients who underwent total arterial revascularization for triple-vessel disease were divided into 2 groups. Group I (n = 188) received a Y-composite graft with 2 arterial conduits, and group II (n = 83) received a Y-composite graft and 1 additional arterial conduit. We retrospectively reviewed the data and compared the clinical results of the 2 groups. RESULTS: The numbers of distal anastomoses were 3.9 +/- 0.7 (mean +/- SD) per patient in group I and 4.1 +/- 0.8 in group II (P = .021). There were 2 in hospital deaths in group I and none in group II (difference not statistically significant). Perioperative myocardial infarction occurred in 2 patients in group I and none in group II (not statistically significant). No patient in either group needed a new intra-aortic balloon pump postoperatively, and no patients experienced hypoperfusion syndrome. A cerebrovascular accident occurred in 1 patient of each group. During the follow-up period (15.9 +/- 6.7 months in group I, 25.2 +/- 8.3 months in group II), 5 patients died (4 in group I, 1 in group II). Two patients in group I and 1 patient in group II needed an intervention on the right coronary artery, but the other patients required no additional procedures and had no symptoms of angina. There were no statistically significant differences in survival and reintervention-free survival between the 2 groups. CONCLUSION: Single Y-composite grafts with 2 arterial conduits may be a safe and effective strategy for treating triple-vessel disease. PMID- 15799910 TI - Surgical treatment of interrupted aortic arch with extraanatomical bypass simultaneous to coronary artery bypass grafting and aortic valve replacement. AB - An interrupted aortic arch accompanied by further surgically reparable cardiac lesions is a rare combination in adult patients. We describe treatment of an interrupted aortic arch, coronary artery bypass grafting (CABG), and aortic valve replacement (AVR) performed simultaneously through median sternotomy in a 64-year old man. The patient underwent surgery performed using standard cardiopulmonary bypass with cannulation of the ascending aorta and the right atrium, hypothermia (24.6degreesC), and blood cardioplegic arrest. Four aortocoronary vein grafts and pericardial aortic valve replacement were carried out. Finally, the posterior pericardium was opened, and a 16-mm prosthesis was anastomosed to the descending aorta during side clamping using a 4-0 monofilament continuous suture. Optimal placement of the prosthesis was obtained by guiding it to the ascending aorta laterally to the right atrium and passing it between the inferior vena cava and right inferior lung vein. The operation was carried out without complications, and the postoperative course was uneventful. Magnetic resonance imaging showed competent aortic valve prosthesis and highly decreased collateral flow via the internal mammary arteries. Postoperatively both inguinal pulses were present, and the patient was free of angina. In the presence of an interrupted aortic arch, extraanatomical bypass via the posterior pericardium between the ascending and descending aorta can safely be performed at the same time as CABG and AVR through a median sternotomy. PMID- 15799911 TI - Monopolar and bipolar radiofrequency ablation surgery: 3-year experience in 90 patients with permanent atrial fibrillation. AB - OBJECTIVE: In our population, permanent atrial fibrillation (pAF) is a serious concomitant problem in patients scheduled for open heart surgery. The high incidence necessitates reliable methods of treating pAF efficiently. We report our 3-year experience with a safe concept of using monopolar and bipolar radiofrequency (RF) ablation procedures. METHODS: Ninety patients (mitral, n = 56; aortic, n = 22; aortic and mitral, n = 1; coronary artery bypass grafting, n = 11) underwent either monopolar (n = 77) or, recently, bipolar (n = 13) RF ablation procedures that produced encircling isolation lesions around the left and the right pulmonary veins (PVs) and a connection line between the two. Amiodarone was given for 3 months after surgery. RESULTS: Hospital mortality was 2.2%. At follow-up, 75% of the patients were in stable sinus rhythm (SR). Preoperative pAF duration, etiology of heart disease, and type of RF energy application were not predictive of the risk of persisting pAF after surgery. Whereas patients (50 of 90) with small preoperative left atrial (LA) diameters (<56 mm) had SR in almost 90% of cases, large preoperative LA diameters (>or=56 mm; 40 of 90 patients) were associated with a significant risk of persisting pAF (P < .05). CONCLUSION: Particularly in cases of small preoperative LA diameters, isolation of the PVs using either monopolar or bipolar RF ablation procedures in combination with amiodarone therapy represents a safe and efficient option for curing pAF in patients undergoing open heart surgery. PMID- 15799912 TI - Congenital fistula between the left internal mammary artery (LIMA) and the pulmonary artery: cause of LIMA bypass occlusion? AB - Congenital fistulas from the left internal mammary artery to the pulmonary artery are rare. We describe a 49-year-old patient with severe aortic valve regurgitation and coronary artery disease. Percutaneous transluminal coronary angioplasty and left anterior descending artery (LAD) stenting had been performed because of a significant proximal LAD lesion. Repeated coronary angiogram 3 months later revealed a patent stent but severe sclerosis up to a 40% stenosis of the LAD after the area of stenting. An aortic valve replacement and a left internal mammary artery (LIMA) bypass to LAD were performed during standard cardiopulmonary bypass (CPB). Because of patient chest pain, a control angiogram was carried out 2 years after surgery and revealed a LIMA-bypass occlusion and a large fistula deriving from the proximal part of the LIMA to the pulmonary artery. The fistula was occluded by coils during an interventional cardiological procedure. Diminished flow in the LIMA bypass due to the fistula in combination with a nonsignificant proximal LAD stenosis are possible reasons for IMA-bypass occlusion. From this case we conclude that angiography of the IMA to detect malformations preoperatively should be mandatory in all cases of arterial coronary revascularization using IMA bypasses. PMID- 15799913 TI - Factors influencing the results of double-valve surgery in patients with fulminant endocarditis: the importance of valve selection. AB - BACKGROUND: Extension of infection below the aortic valve is a serious complication, especially with mitral valve involvement. Mortality is substantial and reinfection can strongly influence outcome. PATIENTS: Of 327 surgical patients with active infective aortic valve endocarditis admitted to the Deutsches Herzzentrum Berlin for surgical treatment between December 1996 and December 2003, 108 had root abscess, and 53 (25.5%) had diagnoses of secondary infective mitral valve disease (SMVD). The mean age (+/-SD) was 53 +/- 14.2 years; there were 37 men and 16 women. METHODS: The secondary lesion on the mitral valve was classified as SMVD requiring double-valve surgery (DVS). This prospective clinical and echocardiographic study revealed 2 paths of infection extension into the mitral valve. In the DVS group, 38 patients (71.7%) had tissue metastatic lesions, and 15 patients (28.3%) had a jet lesion on the mitral valve. Most patients (42) with SMVD had an aortic ring abscess as the primary lesion. RESULTS: All patients with destructive endocarditic doublevalve disease received aortic and mitral valve surgery. In 19 cases (35.8%), mitral valve reconstruction was undertaken; in 4 cases, mitral valve replacement had to be carried out after attempted mitral valve reconstruction. Concomitant mitral valve replacement because of severe damage to the valvular and subvalvular apparatus was performed in 30 patients (56.6%). Other types of surgery performed in 11 cases (20.8%) were 8 closures of a septic ventricular septal defect and 3 closures of a fistula to the right ventricle or right atrium. Twenty-seven patients were treated with a Shelhigh prosthesis, 18 were treated with double-valve replacement (both Shelhigh), and 9 were treated with an aortic Shelhigh prosthesis and concomitant mitral valve reconstruction. Homografts were used in 17 patients, with mitral valve reconstruction carried out in 10 patients and a stented mitral prosthesis in 7. In 9 cases, 2 stented valve prostheses were used. There were 14 early (60 days) deaths (26.4%). Septic shock, severe annular and subannular destruction, and poor left ventricular function (end-diastolic dimension >65 mm, ejection fraction <40%) were the significant risk factors determined in the multivariate analysis. Function of Implants: Continuous and Color Doppler Investigation: Comparative studies of 2 different implants in the aortic position were performed late postoperatively (325 +/- 251 days) for homografts and the Shelhigh stentless prosthesis. The calculated instantaneous (maximal Doppler) gradient and the mean pressure gradient through the aortic implants were 19 +/- 10.4 mm Hg and 12 +/- 5.7 mm Hg, respectively, for the homografts and 24 +/- 8.4 mm Hg and 15 +/- 4.6 mm Hg, respectively, for the Shelhigh stentless prosthesis (not significantly different for the 2 groups). There was no mitral or aortic valve dysfunction. A trivial paravalvular leakage in the mitral position in 1 patient and a pseudoaneurysm of the left ventricular out- flow tract without leakage or valvular dysfunction in another were diagnosed by postoperative Doppler investigation. CONCLUSIONS: The mortality in patients with destructive endocarditis requiring DVS depends mostly on the patients' preoperative hemodynamic situation. The risk of reinfection can be minimized if valve substitutes are properly selected (homografts, Shelhigh No-React SuperStentless and No-React BioConduit in the aortic position, or Shelhigh BioMitral in the mitral position). Concomitant mitral valve reconstruction procedures do not increase the risk of mitral reinfection. PMID- 15799914 TI - Endoscopic radial artery harvest for coronary artery bypass grafting: initial clinical experience. AB - BACKGROUND: Recently, an endoscopic technique was developed to harvest the radial artery (RA) via a 3-cm wrist incision in patients undergoing coronary artery bypass graft (CABG). The aim of this study was to evaluate our initial clinical experience with this technology. METHODS: Data were prospectively collected on 75 consecutive patients undergoing CABG with endoscopic RA harvest using the Ultra Retractor (CardioVations, Somerville, NJ, USA) and the harmonic scalpel (Ethicon Endo-Surgery, Cincinnati, OH, USA). RESULTS: There were 66 men (88%) and 9 women (12%) with a mean age of 60 years (range, 31-77 years). Forty-eight (64%) of cases had non-elective surgery. Thirty-one (41%) of patients had diabetes. Sixty seven percent of the operations were performed on pump and 33% off pump. Average RA harvest time was 66 minutes (range, 25-120 minutes) with a significant learning curve (75 minutes for the first 20 cases and 63 minutes for the last 50 cases). Two (2.6 %) radial arteries were discarded, one because of extensive calcifications and the other because of damage to the conduit. No patients suffered death, perioperative myocardial infarction, or stroke. There was one reexploration of the forearm for a tunnel hematoma. Follow-up was achieved in 100% of patients and averaged 3.6 months (range, 0.5-13 months). Two patients died during the follow-up. There were no myocardial infarctions or reinterventions, with 96% of patients in functional class I /II. There were no motor deficits. There were no sensory deficits in the distribution of the lateral antebrachial cutaneous nerve, but transient mild dorsal thenar numbness or paresthesias were observed in 86% of patients. Cosmetic results were defined as good to excellent in 82% of patients. CONCLUSIONS: Short-term results of endoscopic RA harvest are excellent. There is a significant learning curve. Longterm follow-up as well as structural and functional assessments of the conduit are indicated. PMID- 15799915 TI - Autologous bone marrow-derived stem cell therapy in combination with TMLR. A novel therapeutic option for endstage coronary heart disease: report on 2 cases. AB - We report 2 cases in which patients with coronary heart disease not amenable for conventional revascularization underwent transmyocardial laser revascularization (TMLR) and implantation of AC133+ bone-marrow stem cells. The reason for using TMLR in combination with stem cell application is to take advantage of the synergistic angiogenic effect. The local inflammatory reaction induced by TMLR should serve as an informational platform for stem cells and may trigger their angiogenic differentiation. Functional analysis of myocardial performance after treatment in these 2 cases revealed dramatic improvement of the wall motion at the site of the TMLR and stem cell application. Because TMLR does not enhance myocardial contractility and there was no angiographic evidence of major collaterals to the ischemic region in either patient, we assume that the synergistic effect of stem cells and TMLR-induced angiogenesis occurred; however, our assumption is of a speculative nature. We think that TMLR in combination with stem cell transplantation might become a novel revascularization therapy for ischemic myocardium. PMID- 15799916 TI - Surgical ventricular restoration in end-stage ischemic cardiomyopathy patients. AB - BACKGROUND: Surgical ventricular restoration (SVR) has generally been contraindicated in patients with an ejection fraction (EF) <20%, with pulmonary arterial (PA) pressure >60 mm Hg, and being treated with inotropic agents. PATIENTS AND METHODS: The patients in this study were 6 men and 5 women 50 to 78 years of age (mean, 62.4 years). Three patients were in New York Heart Association (NYHA) class III with an EF <30%. Eight patients were in NYHA class IV with EF <20%, PA pressure >70 mm Hg, and left ventricular asynergy. Three patients had had recent myocardial infarction (MI) with shock and multiple organ failure. Three patients had mitral regurgitation, 1 patient had ventricular septal defect (VSD), 4 patients had diabetes mellitus, and 5 had morbid obesity. All patients underwent intraoperative transesophageal echocardiography and were being treated with milrinone or nesiritide. Seven patients had intraaortic balloon pumps. All patients underwent coronary artery bypass (CAB), receiving 1 to 5 (average, 3.54) grafts per patient. The SVR (Dor) procedure was performed with a Chase Mannequin device. Preoperative end-diastolic volume was 240 to 330 mL, and postoperative volume decreased to 110 to 130 mL. Two patients underwent mitral valve repair, and 1 underwent VSD closure. One patient underwent microwave ablation for atrial fibrillation. RESULTS: Ten (91%) of 11 patients were discharged home in 10 to 14 days. There was 1 death: A 78-year-old man with acute MI died 43 days later of septic shock due to hemodialysis. CONCLUSION: End-stage ischemic cardiomyopathy patients with EF <20% can safely undergo surgery after meticulous preoperative preparation to decrease PA pressure, pulmonary capillary wedge pressure, and peripheral vascular resistance and to increase CO by SVR, CAB, and correction of associated lesions. Mortality was 9% with improved hemodynamics and relief of congestive heart failure in all survivors for 3 to 12 months. PMID- 15799917 TI - Analysis of platelet function during left ventricular support with the Incor and Excor system. AB - Improvements in pump technology and the scarcity of donor organs have led to an increased use of mechanical assist devices, but the problem of thromboembolism has still not been solved. We report on our initial experience with sequentially analyzing platelet function in patients provided with the Incor left ventricular assist device (LVAD) and the Excor LVAD system. Thirteen patients 5 to 61 years old with acute or end-stage heart failure were included in a pilot study. Five of the 10 Incor patients underwent LVAD placement under emergency conditions, and 5 were electively scheduled for surgery. All 3 patients with an Excor device had been connected to an extracorporeal membrane oxygenation system prior to insertion of the device. An anticoagulation protocol including heparin, aspirin, and clopidogrel was employed, and the patients were closely monitored with a special platelet analyzing system that allows dose optimization for antiplatelet drugs. Initial platelet function was normal in only 2 patients (15%). During a follow-up period of 1770 days (cumulative >4.8 years), no early (<30 days) bleeding complications related to device implantation occurred. Late cerebral thromboembolic events were noted in 3 patients. One patient experienced severe stroke mandating neurosurgery during mechanical assist, and 1 patient experienced systemic embolism. The PAP platelet analyzer offers a cheap and reliable alternative to the more expensive thromboelastography method for adequately surveying the efficacy of aspirin and clopidogrel treatment, even if late thromboembolic events cannot be prevented. PMID- 15799918 TI - Virchow triad, but not use of an aortic connector device, predicts early graft failure after off-pump coronary bypass. AB - The risk of early thrombosis of coronary artery bypass (CAB) grafts may be increased after off-pump CAB, particularly after the use of an aortic connector device (ACD) to perform the proximal aortosaphenous anastomosis. We have been investigating tools that quantify the risk of early vein graft thrombosis after off-pump CAB on the basis of the Virchow triad of hypercoagulability, endothelial disease, and low conduit flow. These tools were applied in this prospective trial of a heterogeneous group of patients with varying degrees of aortic disease and who received the Symmetry ACD. After controlling for the Virchow risk factor triad, we hypothesized that the ACD does not independently influence graft thrombosis. There was no statistically significant difference in the early thrombosis rates of vein grafts connected with an ACD versus hand-sewn grafts (6.7% versus 6.5%). We found a wide range of graft flow measurements, platelet function, and vein endothelial phenotypes in patients with traditionally hand sewn vein grafts and with grafts connected with an ACD in patients with a wide range of ascending aortic atherosclerosis. The perioperative combination of platelet hyperreactivity, marginal graft flow, and endothelial disease proved to be highly predictive of early graft failure as seen with postoperative computed tomographic angiography. PMID- 15799919 TI - Effects on reperfusion injury of adding diltiazem to tepid blood cardioplegia. AB - BACKGROUND: Although the present techniques of myocardial preservation for limiting ischemia/reperfusion injury in open heart operations yield excellent results for most patients, certain subgroups of patients with advanced coronary artery disease present a challenge in terms of intraoperative safety. METHODS: In a prospective, randomized, controlled study, we assessed the myocardial protective effects of a total dose of 150 +/- 150 = 300 microg/kg diltiazem added to induction and terminal (reperfusion) doses of tepid blood cardioplegia. We determined the myocardial morphological (ultrastructural) and enzymatic (serum assays for the cardiospecific isoenzyme of creatine kinase [CK-MB]) changes and functional recovery (atrioventricular [AV]-node recovery time and postoperative need for inotropic support) in patients undergoing elective coronary artery bypass operations. The determinations were made with respect to values for control patients, who received the same cardioplegia but without the addition of diltiazem. RESULTS: The mean isoenzyme CK-MB levels and semiquantitative ultrastructural score values of the diltiazem group were significantly less than those of the control group. Although AV-node recovery time was significantly prolonged (P < .05), this factor did not have major clinical impact. CONCLUSIONS: We concluded that the addition of 150 +/- 150 microg/kg diltiazem to the induction and terminal doses of tepid cardioplegia enhanced myocardial protection in elective aortocoronary bypass surgery in high-risk patients and presented no significant additional operative risk. PMID- 15799920 TI - Experience on the way to totally endoscopic atrial septal defect repair. AB - BACKGROUND: Remote-access perfusion and robotics have enabled totally endoscopic closure of atrial septal defect (ASD) and patent foramen ovale (PFO). We report on a stepwise approach to a totally endoscopic procedure. METHODS: Seventeen patients (median age, 39 years; range, 21-55 years) underwent limited-access ASD or PFO closure. As a preparative step, the operation was carried out through minithoracotomy in 11 patients. In parallel, experience with robotic surgery was gained with totally endoscopic coronary artery bypass grafting procedures. After performance of ASD closures in dry-laboratory models using the da Vinci telemanipulation system, 6 patients were operated on in a totally endoscopic fashion. RESULTS: With the endoscopic approach, significant learning curves were noted for cardiopulmonary bypass time y(min) = 226 - 41 * ln(x) (P = .03) and aortic cross-clamp time y(min) = 134 - 42 * ln(x) (P = .01) (x = number of procedures). There was no hospital mortality, and no residual shunts were detected at postoperative echocardiography. Median ventilation time was 9 hours (range, 0-18 hours) for the minithoracotomy group and 6 hours (range, 4-19 hours) for the totally endoscopic group. Median intensive care unit stay was 20 hours (range, 18-24 hours) and 18 hours (range, 18- 120 hours), respectively. CONCLUSIONS: The implementation of robotic totally endoscopic closure of ASD or PFO in a heart surgery program seems to be safe. An intermediate step of performing the operations through minithoracotomy, adapting to remote access perfusion systems, and gaining experience in other robotic cardiac surgical procedures seems worthwhile. Learning curves are apparent, and adequate defect closure does not seem to be compromised by the totally endoscopic approach. PMID- 15799921 TI - First report of no-react bovine internal mammary artery performance and patency. AB - BACKGROUND: No-React treatment is known to render tissues resistant to calcific degeneration and to reduce early inflammatory response. No-React bovine internal mammary artery (NR-IMA) is available for restricted use in Europe. In this first study, our aim was to use magnetic resonance imaging (MRI) to investigate the clinical performance and patency rates of this conduit. METHODS: Seven patients received 8 grafts with NR-IMA. Approval from the Medical Devices Agency of the United Kingdom was obtained for use of this material. One patient needed salvage coronary artery bypass grafting (CABG). Graft patency was investigated with cardiac MRI. One patient was excluded from the MRI study because of the presence of intracerebral metal clips. The mean follow-up period was 2.5 years with a range of 1 to 4.5 years. RESULTS: There was no mortality in this group. After treatment 6 patients were asymptomatic, and 1 patient had class II anginal symptoms. Four (57%) of the 7 NR-IMA grafts remained patent. The longest patency was 4.5 years in a patient who underwent salvage CABG. Other associated grafts in this cohort of patients were 5 left internal mammary arteries (all patent), 1 radial artery graft (patent), and 7 saphenous vein grafts (4 [57%] of 7 patent). There were no occluded NR-IMA grafts in a patient with patent vein grafts. CONCLUSION: We concluded that at 2.5 year follow-up, NR-IMA had a patency rate of 57% (4 of 7 cases). This rate matched the vein graft patency rate in this cohort of patients. With the longest patency of 4.5 years, use of NR-IMA seems to hold promise for the future. PMID- 15799922 TI - The role of arterial grafts in coronary artery bypass grafting: an update. AB - Based on the legacy of the success of the internal thoracic artery as a bypass conduit for myocardial revascularization, the effort to achieve total arterial revascularization continued to evolve in the last 2 decades. Arterial conduits were used as alternatives to saphenous vein for coronary artery bypass grafting. In this review we attempted to assess the success and failure of these conduits and the current trend in myocardial revascularization. PMID- 15799923 TI - Adult patent ductus arteriosus: successful surgical therapy in a rare presentation of a missed finding. AB - Delayed clinical presentation of patent ductus arteriosus (PDA) in adults is very rare. The clinical presentation in adults consists of either aortic or pulmonary valve endocarditis. We report the case of 34-year-old patient with a known history of rheumatic heart disease in childhood and chronic heart failure for 5 years who presented with acute heart failure and no evidence of PDA at prior echocardiography. Blood cultures grew Staphylococcus epidermidis, and echocardiography showed infective endocarditis of both aortic and pulmonary valves in the context of a large and severely inflamed PDA. Cardiopulmonary bypass and deep hypothermic circulatory arrest were used to interrupt the PDA from within the pulmonary artery because of inability to ligate the severely inflamed and calcified PDA prior to bypass. Combined pulmonary and aortic valve replacement and high-dose inotropic support were used. Dialysis was used for renal failure. The patient recovered and was discharged 10 days postoperatively. Six months later the patient was asymptomatic, and renal failure had resolved. The history and surgical management are discussed with an updated review of the literature. PMID- 15799924 TI - Valvuloplasty for anterior mitral leaflet prolapse due to infective endocarditis: report of 2 cases. AB - We report 2 clinical cases of successful repair of an infected mitral valve with a broad-range prolapse of the anterior mitral leaflet (AML) with large vegetation. The chordal transfer technique was used in both cases, and autologous pericardium was transplanted in one case. There was neither residual mitral regurgitation nor recurrence of mitral valve infection. Chordal transfer is a useful technique for AML lesions of infective endocarditis if the corresponding area of the posterior ML remains normal, and even in the case of a broadly destroyed lesion of the AML, it is valuable to try to apply this technique with an autologous pericardial patch. PMID- 15799925 TI - Open versus endo: early experience with endovascular abdominal aortic aneurysm repair beyond the clinical trials. AB - OBJECTIVE: To analyze and compare open (OR) versus endovascular (EVAR) abdominal aortic aneurysm repair at our institution. METHODS: EVAR was attempted in 256 patients at the University of Tennessee Medical Center, Knoxville, between December 1999 and November 2002. One hundred forty patients underwent attempted EVAR, and 116 underwent OR. All patients were included on an intent-to-treat basis, and results were reviewed retrospectively. Statistical methods included the Student t test and chi-square analysis. RESULTS: Patients were age matched between the 2 groups (70.2 years versus 69.0 years; P = .936). Patients in the OR group had significantly higher American Society of Anesthesiologists classes than the EVAR group (2.96 versus 3.07; P = .006). However, there was no difference between the groups, OR versus EVAR, with respect to the presence of chronic obstructive pulmonary disease (55% versus 46%; P = .129), coronary artery disease (69% versus 66%; P = .638), diabetes mellitus (12% versus 18%; P = .167), mean left ventricular ejection fraction (51.8% versus 53.9%; P = .28), or mean preoperative creatinine level (1.2 mg/dL versus 1.1 mg/dL; P = .167). Tobacco use was more prevalent in the OR group (78.4% versus 64.2%; P = .013), and known carotid artery disease was more prevalent in the EVAR group (20.0% versus 6.9%; P = .003). The EVAR group had significantly shorter lengths of stay ( 4.2 versus 9.0 days; P = .000), intensive care unit days (0 versus 3.2; P = .000), time in the operating room (119.6 minutes versus 225.7 minutes; P = .000), and estimated blood loss (189.1 mL versus 897.9 mL; P = .000). Mean aneurysm size was larger in the OR group (5.6 cm versus 4.9 cm; P = .000). Perioperative complications occurred in 31 patients in the OR group and 5 in the EVAR group (P = .000). Two perioperative deaths occurred in the OR group and none in the EVAR group. As of this writing there has been no significant difference in all-cause mortality in the 2 groups (OR 9.6% versus EVAR 8.0%; P = .651). Seven patients in the EVAR group needed secondary interventions. Six were managed with endovascular techniques, and 1 underwent femoral-femoral bypass. CONCLUSIONS: Patients who undergo EVAR have significantly less morbidity and mortality in the perioperative period than do equally matched patients undergoing open repair. In midterm follow up (2-5 years), mortality is no different. Morbidity conferred by the need for secondary intervention in the endovascular group is minimal. PMID- 15799926 TI - Early regional assessment of LV mass regression and function after stentless valve replacement: comparative randomized study. AB - Early regional performance and hypertrophy regression after stentless aortic valve replacement are still incompletely characterized. We compared early postoperative changes of segmental thickness and function after stentless and stented aortic valve replacement as assessed by cardiac magnetic resonance (CMR). In 16 patients randomly assigned to stented (Mosaic, 8 patients) and stentless (Freestyle, 8 patients) groups, 4 parallel short-axis images at the level of the apex (slice 4), midventricle (slices 2-3), and mitral valve (slice 1) were obtained with a 1.5 T CMR scanner (Magnetom Sonata, Siemens) before and 1 month after surgery. Cine images were obtained using an echo gradient sequence. Left ventricle mass was calculated as the difference between the left ventricular end diastolic volume at the epicardial and endocardial borders multiplied by a myocardium density factor (1.05). Each slice was divided into 8 segments (octants) from anterior (octant I-II) to septal (octant V-VIII). A total of 32 segments encompassed the entire heart. From each of these elements end diastolic thickness and systolic function (fractional thickening) were calculated. In stentless valves significant reduction of septal octant thickness on the midventricular slice was noted. There was no difference in regional systolic function-segment thickening. In stented valves no segmental thickness changes were observed. In stentless valves there was early postoperative thickness reduction of septal segments at the midventricular level. However, this finding did not coincide with changes in segmental function. PMID- 15799927 TI - Inferior partial sternotomy for surgical closure of isolated ventricular septal defects in children. AB - BACKGROUND: Surgical closure of isolated ventricular septal defect (VSD) through partial inferior sternotomy offers the advantages of a much shorter, cosmetically superior skin incision, potentially improved sternal stability, a lower rate of infection, and less postoperative pain. We report our technique and results of use of inferior partial sternotomy for closure of isolated VSD in children. PATIENTS AND METHODS: From July 2002 to July 2003, 24 consecutive patients with a median age of 4.5 months (range, 1 month-4.5 years) underwent partial inferior sternotomy for isolated VSD closure. The length of the incision ranged from 4 to 6 cm. Special features of the approach included T incision of the lower sternum (from the fourth intercostal space to the xiphoid), establishment of cardiopulmonary bypass with central cannulation, aortic cross-clamping, and cardioplegic arrest. All VSDs were approached through right atriotomy. Perimembranous VSDs were exposed after detachment of the anterior leaflet of the tricuspid valve and were closed with a continuous suture. Muscular VSDs were approached directly. Perioperative and postoperative echocardiographic findings were available for all patients. Follow-up was complete. RESULTS: There was no mortality or significant surgical morbidity. Median cross-clamping and cardiopulmonary bypass times were 43 and 103 minutes, respectively. All patients were in sinus rhythm. Perioperative and postoperative echocardiography confirmed the absence of any residual defects in perimembranous VSDs and the presence of a trace residual VSD in 4 patients with muscular VSDs. Optimal healing of the partial sternotomy was obtained in all patients. CONCLUSIONS: Inferior partial sternotomy is less invasive than and cosmetically superior to full sternotomy. It provides excellent results when applied to isolated VSD with standard surgical techniques. PMID- 15799928 TI - Evaluation of myocardial flow reserve using pharmacological stress thallium-201 single-photon emission computed tomography: is there a difference between total arterial off-pump coronary artery bypass grafting and conventional coronary artery bypass grafting? AB - BACKGROUND: The advantage of total arterial off-pump coronary bypass grafting (OPCAB) over conventional onpump coronary artery bypass grafting with 1 internal thoracic artery and veins (CCAB) in terms of myocardial flow reserve has not been studied. We studied these procedures using thallium- 201 perfusion single-photon emission computed tomography (Tl-201 perfusion SPECT). METHODS: Between 1997 and 2001, 152 patients were recruited from our database (OPCAB, n = 100; CCAB, n = 52). All patients underwent pharmacological stress Tl-201 perfusion SPECT 3 to 12 months after bypass surgery. Myocardial perfusion was analyzed semiquantitatively with a 5-point scoring system in a 20-segment model (0, normal, to 4, absence of uptake). Summed stress (SSS), rest (SRS), and difference score (SDS) of the entire myocardium as well as average scores (ASS, ARS, ADS) of individual walls (anterior, septal, lateral, and inferior) were compared by Student t test as well as by repeated-measures analysis of variance with Bonferroni correction. RESULTS: The SSS, SRS, and SDS of OPCAB versus those of CCAB were 6.86 +/- 0.72 versus 7.17 +/- 0.92, 3.95 +/- 0.57 versus 3.75 +/- 0.73, and 2.91 +/- 0.47 versus 3.42 +/- 0.74 (P > .05). However, the lateral wall showed lower scores in OPCAB (ASS, 0.18 versus 0.41, P = .015; ARS, 0.12 versus 0.20, P = .168; ADS, 0.06 versus 0.21, P = .031). The septal wall had higher scores in OPCAB (ASS, 0.33 versus 0.12, P = .003; ARS, 0.18 versus 0.07, P = .037; ADS, 0.14 versus 0.04, P = .030). The anterior and inferior walls were not different between the 2 groups. CONCLUSIONS: OPCAB led to results similar to those of CCAB. The better results in the lateral wall have been the effect of grafting radial artery rather than vein. The similarity in myocardial reserve in the inferior wall between the 2 groups needs further study. There was no deleterious effect of off-pump as opposed to on pump CAB. PMID- 15799929 TI - The right internal thoracic artery and right gastroepiploic artery: alternative sites for proximal anastomosis in patients with atherosclerotic calcified aorta. AB - BACKGROUND: Atherosclerotic or calcified ascending aorta is an important predictor of adverse cerebrovascular events. Using off-pump coronary artery bypass (OPCAB) with composite and in situ arterial grafting to avoid aortic manipulation and clamping may reduce the risk of stroke related to aortic atheroembolism. When the aorta is calcified and cannot serve as a safe site for proximal anastomosis, this anastomosis can be performed on the proximal segment of the right internal thoracic artery (ITA) and right gastroepiploic artery (RGEA). Four such cases are described. METHODS: In 2 patients, the proximal right ITA was used as the site for proximal saphenous vein graft (SVG) anastomosis. Chronic obstructive lung disease in one patient and insulin-dependent diabetes in the other precluded performance of OPCAB with bilateral ITA. In addition, positive Allen test precluded performance of composite T-graft with radial artery (RA) on ITA. Both SVGs were anastomosed distally to the posterior descending artery. In 2 other patients, RA was connected end-to-side to the proximal segment of the RGEA. Both of these patients had repeat operations. The distal end of the RGEA was too small, and concerns regarding the future flow to a very large coronary bed precluded its use as an in situ graft. RESULTS: All patients underwent the operative procedures without any neurological or cardiovascular adverse effects, and all are midterm, symptom-free survivors. Postoperative graft patency was confirmed intraoperatively with flow measurements and postoperatively with control angiography or coronary imaging computed tomography. CONCLUSIONS: The RITA and proximal RGEA can serve safely as sites for proximal anastomosis in patients with atherosclerotic calcified aorta undergoing OPCAB. PMID- 15799930 TI - Reconstructed bicuspid aortic valve after 10 years: clinical and echocardiographic follow-up. AB - BACKGROUND: Sixteen patients (mean age, 30.9 +/- 12.9 years; range, 9-79 years) with incompetent bicuspid aortic valves underwent valve-sparing correction between 1992 and 1995. METHODS: All patients underwent triangular resection of the enlarged leaflet. In addition, 13 patients underwent commissuroplasty. In 3 patients leaflet perforations were corrected with glutaraldehyde-fixed autologous pericardial patch. All patients underwent annual echocardiographic and clinical examinations at our institution. RESULTS: During follow-up 1 patient died of heart failure, and 3 patients underwent reoperations because of valve incompetence or dilatation of the sinus. Two patients underwent reoperation perioperatively. At long-term follow-up (mean, 10.06 +/- 1.01 years) the remaining 10 patients were in New York Heart Association class I. No patient was receiving anticoagulation, and no thromboembolic, bleeding, or endocarditis events were observed in 107.2 cumulative patientyears of follow-up. Mean grade of regurgitation was 0.7 +/- 0.5; mean aortic flow velocity was 2.29 +/- 0.47 m/s. Optimal valvular function led to normal ventricular diameters (left ventricular end systolic diameter, 39.2 +/- 4.3 mm; left ventricular end diastolic diameter, 56.2 +/- 5.9 mm) and normal ventricular function (fractional shortening, 31.5% +/ 0.1%). The dimensions of the aortic root were stable from 1-year to late followup (mean aortic annulus, 27.1 +/- 6.8 mm; sinus of Valsalva, 33.0 +/- 7.1 mm; sinotubular junction, 34.1 +/- 7.7 mm; ascending aorta, 31.6 +/- 7.4 mm). CONCLUSION: In contrast to early follow-up results of 5 reoperations, clinical and echocardiographic results were excellent for 10 patients who had undergone reconstruction of bicuspid incompetent aortic valves, and the patients were in stable condition after 10 years. However, the mode of early failure is unknown. Reconstruction of bicuspid valves is possible in selected patients. PMID- 15799931 TI - Revascularization of the left anterior descending artery with drug-eluting stents: comparison with arterial off-pump surgery. AB - BACKGROUND: This study compared early results of left anterior descending artery (LAD) stenting using drug-eluting stents (Cypher) with off-pump bilateral internal thoracic arterial (BITA) grafting. METHODS: From June 2002 to June 2003, 200 consecutive patients underwent myocardial revascularization of the LAD territory, 100 by Cypher and 100 by BITA. The 2 groups were similar; however, left main disease and triple-vessel disease (20% and 75% versus 2% and 28%), age >70 (36% versus 17%) and intraaortic balloon pump (7% versus 0%) were more prevalent in the BITA group, and prior percutaneous coronary angiogplasty to the LAD was more prevalent in the Cypher group (28% versus 16%). RESULTS: The number of coronary vessels treated per patient in the BITA group was higher (2.7 versus 1.45, P < .01). Thirty-day mortality was 1% in the BITA group and 0% in the Cypher group. Mean follow-up was 12 months. There was 1 late death in each group. Angina returned in 32% of the Cypher group and in 1% of the BITA group. There were 9 reinterventions in the Cypher group: 7 coronary angioplasties (including 2 to the LAD) and 2 surgical interventions. There was neither recurrent angina nor reintervention in the surgical group. CONCLUSIONS: Despite a higher risk profile of patients treated with BITA, their clinical outcome is better. A longer and more complete angiographic follow-up is required to determine the role of drug eluting stents in LAD revascularization. PMID- 15799932 TI - Total arterial revascularization for multiple vessel coronary artery disease: with or without cardiopulmonary bypass. AB - BACKGROUND: To assess the usefulness of off-pump technique for more technically demanding coronary artery bypass procedures using exclusively arterial conduits. METHODS: Analysis of perioperative data of 324 consecutive patients in whom total arterial revascularization for multiple- vessel coronary artery disease was performed--181 cases on-pump and 143 cases off-pump. RESULTS: On average in the on-pump group 2.7 +/- 0.8 (range, 2-5) grafts per patient were constructed versus 2.4 +/- 0.7 (range, 2-4) grafts per patient in the off-pump group (P < .001). Of the total number of 490 anastomoses performed on-pump, 83 (17%) were side-to-side and of 349 anastomoses performed off-pump, 51(15%) were side-to-side, a nonsignificant difference (P = .4). The aorta was used as a site for proximal anastomosis of 1 or more arterial conduits in 105 patients (58%) who underwent on pump surgery and in 57 patients (40%) who underwent off-pump surgery (P = .002). In the off-pump group, the right internal thoracic artery (RITA) was rarely (12%) routed through the transverse sinus to circumflex branches compared with the on pump group (34%) (P = .017). RITA in off-pump patients was more often used to revascularize the anterior wall (47% versus 29%; P = .08). We observed no difference in mortality (1.7% versus 0%; P = .3), incidence of perioperative myocardial infarction (8.8% versus 7.7%; P = .8), stroke (1.7% versus 1.4%; P = .8), or atrial fibrillation (24% versus 19%; P = .3). We observed less inotropic support and less blood-product use in off-pump patients. CONCLUSION: Total arterial revascularization for multiple-vessel coronary artery disease may be safely performed off-pump. We observed tendency to somewhat smoother postoperative course in the off-pump group. PMID- 15799933 TI - Early clinical experience with a new tubular equine pericardial stentless aortic valve. AB - BACKGROUND: This study details a single-center experience with the 3F Therapeutics stentless aortic bioprosthesis(investigational device) and is part of a prospective,nonrandomized worldwide multicenter study. We assessed the implantation procedure and the valve's efficacy in terms of early mortality and morbidity and early echocardiographic valve performance. METHODS: Between June 2001 and March 2004, 24 patients (14 men/10 women) underwent aortic valve replacement(AVR) with a 3F valve. Mean age was 72 +/- 13 years(range, 31-88 years). Combined revascularization was performed in 12 patients; 1 patient received biatrial ablation therapy, 1 patient a myectomy, and 3 patients combined carotid endarterectomy. Echocardiographic systolic gradient and valve performance were investigated intra- and postoperatively by Doppler echocardiography. RESULTS: There were 2 perioperative deaths, 1 non-valve related, due to aortic rupture in an 83-year-old woman, and 1 fatal cerebral embolism in a 77-year-old woman 5 days postoperatively. Cardiopulmonary bypass time was 102 +/- 32 minutes,aortic cross-clamp time was 79 +/- 24 minutes. Sizes for implanted 3F valves were 5 x 23 mm, 6 x 25 mm, 7 x 27 mm,and 6 x 29 mm. Follow-up systolic gradient results were 11.5 +/- 4.7 mm Hg at 30 days (n = 21), 11.4 +/- 4.5 mm Hg at 6 months (n = 18), and 13.3 +/- 4.4 mm Hg at 12 months (n =13). During the follow-up period trivial central aortic valve regurgitation was found in 6 patients. CONCLUSION: The 3F aortic valve shows favorable preliminary hemodynamic results. Owing to the new valve design,implantation technique is simplified compared with other stentless valves. Anticoagulation treatment is mandatory for the first 3 months postoperatively. Long-term observation is necessary to assess life span and durability. PMID- 15799934 TI - The clinical and fiscal impact of endovascular repair of abdominal aortic aneurysms. AB - PURPOSE: This study analyzed cost, reimbursement, application, and outcome differences between endovascular (ER) and open repair (OR) of abdominal aortic aneurysms (AAA) in a community hospital. METHOD: A total of 187 consecutive elective AAA repairs by both methods (69 ER, 118 OR) performed at a single center between July 2001 and March 2003 were analyzed. Average values in postoperative clinical and fiscal demographics were calculated for this period. RESULTS: The average length of stay was higher for OR than ER (9.38 days versus 1.94 days, P < .001). Significant reductions in operative time (4 hours versus 2.67 hours) and intensive care unit use (100% versus 4.05%) were seen in the ER population. Total hospital costs were slightly higher for OR than ER (21,989 dollars versus 19,668 dollars) despite a considerable difference in cost of the grafts (>11,000 dollars for ER versus <500 dollars for OR). However, hospital charges were much lower for ER than OR (32,660 dollars versus 48,877 dollars), and there was an average loss of 4986 dollars on ER cases versus a profit of 2064 dollars on OR procedures. Thirty-day mortality was not significantly improved in the ER population (1.45% versus 2.54%, P = .05). CONCLUSIONS: Our data suggest that ER offers improvements in hospital convalescence and operating room times but no improvements in fiscal impact or overall morbidity/ mortality rates when similar preoperative medical risks exist. PMID- 15799935 TI - Challenges associated with the integration of endovascular repair of abdominal aortic aneurysms in a community hospital. AB - PURPOSE: There has been considerable debate regarding the proper place for endovascular repair (ER) of abdominal aortic aneurysms (AAAs) versus traditional open repair (OR). Our study compared preoperative patient demographics and outcomes for elective, asymptomatic AAA repairs performed at our center over a 33 month period. METHODS: For this study, we selected 342 consecutive elective infrarenal AAA repairs performed between July 1, 2000, and March 31, 2003, at Riverside Methodist Hospital. The patients underwent either ER or OR, depending on patient and surgeon collaborative determinations. Ruptured and symptomatic AAAs were excluded from our study. Preoperative demographics, anesthesia, complications, and discharge status for the 2 groups were analyzed, and statistical analysis was done to determine statistically significant differences. RESULTS: The preoperative status of the ER and OR patient groups were essentially similar. There were only 3 significant differences between the 2 groups: alcohol use was higher for the OR group than for the ER group (12.0% versus 5.2%; P = .04), and the incidence of type II diabetes mellitus and peripheral vascular disease were lower for the OR group compared with the ER group (6.7% versus 13.4% [P = .04] and 18.3% versus 30.6% [P = .008], respectively). The OR group used more general anesthesia than the ER group (99% versus 86%; P < .001) and had more complications, including dysrhythmia (8.65% versus 1.59%; P = .005), ileus (13.94% versus 0.79%; P < .0001), infection (8.17% versus 0.0%; P = .0007), respiratory complications (12.50% versus 1.59%; P = .0003), and renal complications (5.29% versus 0.79%; P = .032). The ER group had a higher rate of wound hematoma (4.76% versus 0.48%; P = .007). ER patients also had significantly less blood loss (379 mL versus 1930 mL; P < .001), a better independent discharge status (P < .0001), a shorter length of stay (1.8 days versus 8.2 days; P < .001), and a lower mortality rate (0.75% versus 3.85%; P = .0954). CONCLUSIONS: From our study we cautiously continue to encourage the consideration of the ER of AAAs in our patient population while being mindful of its limitations. PMID- 15799936 TI - Oblique groin incision for endovascular repair of abdominal aortic aneurysms. AB - PURPOSE: Groin incision for access to the femoral artery is necessary for stent graft repair of abdominal aortic aneurysm (AAA). However, lymphatic and infectious complications can occur during wound healing after surgery. A vertical incision traditionally is used, but a new oblique incision technique has the potential to reduce complications. We report our results from AAA stent repairs performed via oblique incisions at our center for a 33-month period. METHODS: Data for 134 consecutive patients undergoing elective stent repair of asymptomatic infrarenal AAA performed by 2 cardiovascular surgeons at a single center from July 1, 2000, to March 31, 2003, were gathered for analysis. Wound complication percentages for infections, paresthesias, sepsis, and seroma were calculated. RESULTS: In 134 patients, a total of 278 incisions were made. Reasons for extra incisions included improving catheter entry angle (5 cases), tunneling for vascular tapes (3 cases), and initial approach too low (2 cases). The percentages were, however, calculated on a per-patient basis. Thirty day analysis revealed a 0% infection rate, 4.7% incidence of paresthesias, 2.38% rate of wound seroma, and 0% wound sepsis rate. CONCLUSION: Because of the nonexistent infection rate and low wound complication rate, our data supports the use of oblique groin incisions for stent repair of asymptomatic infrarenal AAA. PMID- 15799937 TI - The role of palmaris muscle tendon in mitral valve annulus reconstruction: a novel technique for mitral valve repair. AB - BACKGROUND: Autogenous tissue for heart valve repair is limited to pericardium and fascia lata. Prosthetic annuloplasty rings have limitations and are expensive. No previous cadaveric study has documented autotransplantation of forearm tendons for mitral valve repair. The purposes of this anatomical study were (1) to determine the feasibility of using tendons for annular reconstruction during mitral valve repair (band or ring shape) and (2) to compare the metric dimensions and gross morphology with those of prosthetic rings. METHODS: The palmaris tendon (PM) (n = 7) was harvested from forearms of human cadavers and prepared in the dissecting room. The tendon was incorporated along the valve annulus. Surgery consisted of mitral valve annuloplasty performed with an undersized and a complete tendon. RESULTS: On gross handling PM tissue was morphologically resilient. Preparation and use of PM for annuloplasty were feasible. Anatomical reconstruction of the annulus with autogenous tissue using this tendon also was feasible. The annulus behaved as a flexible ring that allowed for improved atrioventricular continuity. Complete (n = 7) and partial (n = 7) annuloplasty rings were constructed. Metric dimensions were similar to those of prosthetic rings. CONCLUSION: PM is a suitable novel autogenous tissue that can be harvested together with the radial artery and has direct relevance in ischemic mitral valve regurgitation. These tendons can be easily utilized to achieve results similar to those of pericardial reconstruction and prosthetic annuloplasty rings. The tissue is readily available from the patient, and no extra cost is incurred. Further chemical studies in the human subjects are warranted. PMID- 15799938 TI - New techniques and innovative technology: an integrated element of our cardiovascular and thoracic specialty. PMID- 15799939 TI - Harmonic scalpel for pericardiectomy: novel approach to an old cardiac dilemma. AB - BACKGROUND: Pericardiectomy for constrictive or calcific pericarditis is a technical challenge because of dense adhesions to the epicardial surface of the heart. The procedure is fraught with the possibility of urgent cardiopulmonary bypass from excessive bleeding or cardiac laceration. We propose the use of a harmonic scalpel to perform adhesiolysis with less bleeding and cardiac trauma. METHODS AND MATERIALS: A retrospective review of 7 pericardiectomies performed with a handheld harmonic scalpel over the past 2 years was performed. Requirements for blood products, the need for cardiopulmonary bypass, and mortality were examined. RESULTS: Four of the patients underwent pericardiectomy alone, and 3 patients underwent pericardiectomy with additional cardiac procedures. The 30-day mortality was zero. No patient needed blood transfusions or urgent cardiopulmonary bypass for bleeding. No patient developed malignant arrhythmias. CONCLUSIONS: Use of a harmonic scalpel is a safe and efficient technique for pericardiectomy. Adhesiolysis is less treacherous because of the bloodless operative field; moreover, the harmonic scalpel is not arrhythmogenic. PMID- 15799940 TI - New flexible polymeric heart valve prostheses for the mitral and aortic positions. AB - OBJECTIVE: Current prosthetic heart valves necessitate permanent anticoagulation or have limited durability and impaired hemodynamic performance compared with natural valves. We report in vivo and in vitro results with new polymeric valve prostheses that have a special design for the mitral and aortic positions. The aims are improved durability and elimination of the need for permanent anticoagulation. METHODS: The mitral and aortic prostheses (Adiam Life Science, Erkelenz, Germany) are made entirely of polycarbonate urethane (PCU). The bileaflet asymmetric mitral valve mimics natural, nonaxial inflow, which creates a left ventricular vortex, saving energy for systolic ejection of blood. The trileaflet aortic prosthesis has diminished pressure loss and reduced stress and strain peaks at the commissures. The valves were subjected to long-term in vitro testing and in vivo testing in a growing calf model (20 weeks; 7 mitral and 7 aortic valves) with comparison with 2 commercial bioprostheses (7 mitral, 2 aortic). Two-dimensional echocardiography was performed after implantation and prior to sacrifice with autopsy and valve examination. RESULTS: In vitro durability of the PCU valves was proved up to 20 years. In vivo durability and hemodynamics were superior to those of all bioprostheses. Survival of PCU valves versus bioprostheses was 7 versus 2 mitral valves and 5 versus 0 aortic valves, respectively. Two animals with PCU aortic valves died of pannus overgrowth that caused severe left ventricular outflow tract obstruction without changes in the valves. Degeneration and calcification were mild (mitral) and moderate (aortic) in PCU valves but were severe in biological valves. There was no increased thrombogenicity of the PCU valves compared with bioprostheses. CONCLUSION: The new flexible polymeric aortic and mitral valve prostheses were superior to current bioprostheses in animal testing. PMID- 15799941 TI - Effects on weed and invertebrate abundance and diversity of herbicide management in genetically modified herbicide-tolerant winter-sown oilseed rape. AB - We evaluated the effects of the herbicide management associated with genetically modified herbicide-tolerant (GMHT) winter oilseed rape (WOSR) on weed and invertebrate abundance and diversity by testing the null hypotheses that there is no difference between the effects of herbicide management of GMHT WOSR and that of comparable conventional varieties. For total weeds, there were few treatment differences between GMHT and conventional cropping, but large and opposite treatment effects were observed for dicots and monocots. In the GMHT treatment, there were fewer dicots and monocots than in conventional crops. At harvest, dicot biomass and seed rain in the GMHT treatment were one-third of that in the conventional, while monocot biomass was threefold greater and monocot seed rain almost fivefold greater in the GMHT treatment than in the conventional. These differential effects persisted into the following two years of the rotation. Bees and Butterflies that forage and select for dicot weeds were less abundant in GMHT WORS management in July. Year totals for Collembola were greater under GMHT management. There were few other treatment effects on invertebrates, despite the marked effects of herbicide management on the weeds. PMID- 15799942 TI - Force enhancement and relaxation rates after stretch of activated muscle fibres. AB - The residual force enhancement following muscle stretch might be associated with an increase in the proportion of attached cross-bridges, as supported by stiffness measurements. In this case, it could be caused by an increase in the attachment or a decrease in the detachment rate of cross-bridges, or a combination of the two. The purpose of this study was to investigate if the stretch-induced force enhancement is related to cross-bridge attachment/detachment kinetics. Single muscle fibres dissected from the lumbrical muscle of frog were place at a length approximately 20% longer than the plateau of the force-length relationship; they were maximally activated, and after full isometric force was reached, ramp stretches were imposed with amplitudes of 5 and 10% fibre length, at a speed of 40% fibre length s(-1). Experiments were performed in Ringer's solution, and with the addition of 2, 5 and 10 nM of 2,3 butanedione monoxime (BDM), a drug that places cross-bridges in a pre-power stroke, state, inhibiting force production. The total force following stretch was higher than the corresponding force measured after isometric contraction at the corresponding length. This residual force enhancement was accompanied by an increase relaxation time. BDM, which decreases force production during isometric contractions, considerably increased the relative levels of force enhancement. BDM also increased relaxation times after stretch, beyond the levels observed during reference contractions in Ringer's solution, and beyond isometric control tests at the corresponding BDM concentrations. Together, these results support the idea that force enhancement is caused, at least in part, by a decrease in cross-bridge detachment rates, as manifested by the increased relaxation times following fibre stretch. PMID- 15799943 TI - Hamilton and Zuk meet heterozygosity? Song repertoire size indicates inbreeding and immunity in song sparrows (Melospiza melodia). AB - Hamilton and Zuk's influential hypothesis of parasite-mediated sexual selection proposes that exaggerated secondary sexual ornaments indicate a male's addictive genetic immunity to parasites. However, genetic correlated of ornaments and immunity have rarely been explicitly identified. Evidence supporting Hamilton and Zuk's hypothesis has instead been gathered by looking for positive phenotypic correlations between ornamentation and immunity; such correlations are assumed to reflect causal, addictive relationships between these traits. We show that in a song sparrows, Melospiza melodia, male's song repertoire size, a secondary sexual trait, increased with his cell-mediated immune response (CMI) to an experimental challenge. However, this phenotypic correlation could be explained because both repertoire size and CMI declined with a male's inbreeding level. Repertoire size therefore primarily indicated a male's relative heterozygosity, a non-addictive genetic predictor of immunity. Caution may therefore be required when interpreting phenotypic correlations as support for Hamilton and Zuk's addictive model of sexual selection. However, our results suggest that female song sparrows choosing with large repertoires would on average acquire more outbred and therefore more heterozygous mates. Such genetic dominance effects on ornamentation are likely to influence evolutionary trajectories of female choice, and should be explicitly incorporated into genetic models of sexual selection. PMID- 15799944 TI - Orientation in the wandering albatross: interfering with magnetic perception does not affect orientation performance. AB - After making foraging flights of several thousands of kilometers, wandering albatrosses (Diomedea exulans) are able to pinpoint a specific remote island where their nests are located. This impressive navigation ability is highly precise but its nature is mysterious. Here we examined whether albatrosses rely on the perception of the Earth's magnetic field to accomplish this task. We disturbed the perception of the magnetic field using mobile magnets glued to the head of nine albatrosses and compared their performances with those of 11 control birds. We then used satellite telemetry to monitor their behavior. We found that the ability of birds to home specific nest sites was unimpaired by this manipulation. In particular, experimental and control birds did not show significant differences with respect to either foraging trip duration, or length, or with respect to homing straightness index. Our data suggest that wandering albatrosses do not require magnetic cues to navigate back to their nesting birds. PMID- 15799945 TI - Low effective population size and evidence for inbreeding in an overexploited flatfish, plaice (Pleuronectes platessa L.). AB - Overexploitation and subsequent collapses of major worldwide fisheries has made it clear that marine stocks are no inexhaustible. Unfortunately, the perception remains that marine fished are resilient to large population reductions, as even a commercially 'collapsed' stock will still consist of millions of individuals. Coupled with this notion is the idea that fisheries can, therefore, have little effect on the genetic diversity of stocks. We used DNA from archived otoliths collected between 1924 and 1972 together with 2002 juvenile;s tissue to estimate effective population size (Ne) in plaice (Pleuronrctes platessa). Ne was estimated at 20,000 in the North Sea and 2000 in Iceland. These values are five orders of magnitude smaller than the estimated census size foe the two locations. Populations examined between 1924 and 1960 were in Hardy-Weinberg equilibrium, whereas populations examined after 1970 were not. Extensive testing was performed to rule out genotyping artefacts and Wahlund effects. The significant heterozygote deficiencies found from 1970 onward were attributed to inbreeding. The emergence of inbreeding between 1905 and 19070 coincides with the increase in fishing mortality after World War II. Although the biological mechanisms remain speculative, our demonstration of inbreeding signals the need for understanding the social and mating behaviour in commercially important fishes. PMID- 15799946 TI - A parasite-mediated life-history shift in Daphnia magna. AB - The impact of parasitism on host populations will be modulated by both genetic variation for susceptibility, and phenotypically plastic-life-history traits that are altered to lessen the fitness consequences of infection. In this study we tested for life-history shifts in the crustacean Daphnia magna following exposure to the horizontally transmitted microsporidian, Glugoides intestinalis. In two separate experiments, we exposed hosts to parasite spores and measured their fecundity relative to controls. We show that host exposed G. intestinalis show fecundity compensation, i.e. hosts shift their life-history strategy towards early production. Our experiments included multiple host genotypes, and subtle differences among them indicated that fecundity compensation could be subject to parasite-mediated natural selection. PMID- 15799947 TI - Host cell preference and variable transmission strategies in malaria parasites. AB - Malaria and other haemosporin parasites must undergo a round of sexual reproduction in their insect vector in order to produce stages that can be transmitted to vertebrate hosts. Consequently, it is crucial that parasites produce the sex ratio (proportion of male sexual stages) that will maximize the number of fertilization and thus, transmission to new vertebrate hosts. There is some evidence to show that, consistent with evolutionary theory, the sex ratios of malaria parasites are negatively correlated to their inbreeding rate. However, recent theory has shown that when fertilization success is compromised, parasites should respond by increasing their investment in sexual stages or by producing a less female biased ration than predicted by their inbreeding rate alone. Here, we show that two species of rodent malaria, Plasmodium chabaudi and Plasmodium vinckei petteri, adopt different strategies in response to host anaemia, a factor though to compromise transmission success: P. chabaudi increases investment in sexual stages, whereas P. vinckei produces a less female biased sex ratio. We suggest that these different transmission strategies may be due to marked differences in host cell preference. PMID- 15799948 TI - Comparison of climate space and phylogeny of Marmota (Mammalia: Rodentia) indicates a connection between evolutionary history and climate preference. AB - Palaeobiologists have investigated the evolutionary responses to extinct organisms to climate change, and have also used extinct organisms to reconstruct palaeoclimates. There is evidence of a disconnection between climate change and evolution that suggest that organism may not be accurate paleoclimate indicators. Here, marmots (Marmota sp.) are used as a case study to examine whether similarity of climate preferences is correlated with evolutionary relatedness of species. This study tests for a relationship between phylogenetic distance and 'climate distance' of species with a clade. There should be a significant congruence between maximus likelihood distance and standardized Euclidian distance between climates if daughter species tend to say in environments similar to parent species. Marmots make a good test case because there are many extant species, their phylogenetics are well established and individual survival is linked to climatic factors. A Mantel test indicates a significant correlation between climate and phylogenetic distance matrices, but this relationship explains only a small fraction of the variance (regression R(2) = 0.114). These results that (i) closely related species of marmots tend to stay in similar environments; (ii) marmots may be more susceptible than may mammals to global climate change; and (iii) because of the considerable noise in this system, the correlation cannot be used for detailed palaeoclimate reconstruction. PMID- 15799949 TI - Mitochondria as integrators of information in an early-evolving animal: insights from a triterpenoid metabolite. AB - Mitochondria have the capacity to integrate environmental signals and, in animals with active stem cell populations, trigger responses in terms of growth and growth form. Colonial hydroids, which consists of feeding polyps connected by tube-like stolons, were treated with avicis, triterpenoid electrophiles whose anti-cancer properties in human cells are mediated in part by mitochondria. In treated hydroids, both oxygen uptake and mitochondrial reactive oxygen species were diminished relative to controls, similar to that observed in human cells exposed to avicins. While untreated colonies exhibit more stolon branches and connections in the centre of the colony than at the periphery, treated colonies exhibit the opposite: fewer stolon branches in the centre of the colony than at the periphery. The resulting growth form suggest an inversion of the normal pattern of colony development mediated by mitochondrial and redox-related perturbations. An as-yet-uncharacterized gradient within the colony may determine the ultimate phenotypic effects of avicin perturbation. PMID- 15799950 TI - Phenotypic plasticity in the developmental integration of morphological trade offs and secondary sexual trait compensation. AB - Trait exaggeration through sexual selection will tale place alongside other changes in phenotype. Exaggerated morphology might be compensated by parallel changes in traits that support, enhance or facilitate exaggeration: 'secondary sexual trait compensation' (SSTC). Alternatively, exaggeration might be realized at the expense of other traits through morphological trade-offs. For the most part, SSTC has only been examined interspecifically. For these phenomena to be important intraspecifically, the sexual trait must be developmentally integrated with the compensatory or competing trait. We studied developmental integration in two species with different development: the holometabolous beetle Onthophagus taurus and the hemimetabolous earwig Forficula auricularia. Male-dimorphic variation in trait exaggeration was exploited to expose both trade-offs and SSTC. We found evidence for morphological trade-offs in O. taurus, but no F. auricularia, supporting the notion that trade-offs are more likely in closed developmetal systems. However, we found these trade-offs were not limited solely to traits growing close together. Developmental integration of structures involved in SSTC were detected in both species. The developmental integration of SSTC was phenotypically plastic, such that the compensation for relatively larger sexual traits was greater in the exasperated male morphs. Evidence of intraspecific SSTC demands studies of the selective, genetic and developmental architecture of phenotypic integration. PMID- 15799951 TI - Among- and within-patch components of genetic diversity respond at different rates to habitat fragmentation: an empirical demonstration. AB - Habitat fragmentation is a ubiquitous by-product of human activities that can alter the genetic structure of natural populations, with potentially deleterious effects on population persistence and evolutionary potential. When habitat fragmentation results in the subdivision of a population, random genetic drift then leads to the erosion of genetic diversity from within the resulting subpopulation, random genetic drift then leads to the erosion of genetic diversity from within the resulting subpopulations and greater genetic divergence among them. Theoretical and simulation analyses predict that these two main genetic effects of fragmentation, greater differentiation among resulting subpopulation and reduced genetic diversity within them, will proceed at very different rates. Despite important implications for the interpretation of genetics data from fragmented populations, empirical evidence for this phenomenon has been lacking. In this analysis, we carry out an empirical study in population of an alpine meadow-dwelling butterfly, which have become fragmented increasing forest cover over five decades. We show that genetic differentiation among subpopulations (G(ST)) is most highly correlated with contemporary forest cover, while genetics diversity within subpopulation (expected heterozygosity) is better correlated with the spatial pattern of forest cover 40 years in the past. Thus, where habitat fragmentation has occurred in recent decades, genetic differentiation among subpopulation can be near equilibrium while contemporary measures of within subpopulation diversity may substantially overestimate the equilibrium values that will eventually be attained. PMID- 15799952 TI - The consequences of Zipf's law for syntax and symbolic reference. AB - Although many species possess rudimentary communication systems, humans seem to be unique with regard to making use of syntax and symbolic reference. Recent approaches to the evolution of language formalize why syntax is selectively advantageous compared with isolated signal communication systems, but do not explain how signals naturally combine. Even more recent work has shown that if a communication system maximizes communicative efficiency while minimizing the cost of communication, or if a communication system constrains ambiguity in a non trivial way while a certain entropy is maximized, signal frequencies will be distributed according to Zipf's law. Here we show that such communication principles give rise not only to signals that have many traits in common with the linking words in real human languages, but also to a rudimentary sort of syntax and symbolic reference. PMID- 15799953 TI - Evaluating real-life high-throughput screening data. AB - High-throughput screening (HTS) is the result of a concerted effort of chemistry, biology, information technology, and engineering. Many factors beyond the biology of the assay influence the quality and outcome of the screening process, yet data analysis and quality control are often focused on the analysis of a limited set of control wells and the calculated values derived from these wells. Taking into account the large number of variables and the amount of data generated, multiple views of the screening data are necessary to guarantee quality and validity of HTS results. This article does not aim to give an exhaustive outlook on HTS data analysis but tries to illustrate the shortfalls of a reductionist approach focused on control wells and give examples for further analysis. PMID- 15799954 TI - Rescue and in situ selection and evaluation (RISE): a method for high-throughput panning of phage display libraries. AB - Phage display has proven to be an invaluable instrument in the search for proteins and peptides with optimized or novel functions. The amplification and selection of phage libraries typically involve several operations and handling large bacterial cultures during each round. Purification of the assembled phage particles after rescue adds to the labor and time demand. The authors therefore devised a method, termed rescue and in situ selection and evaluation (RISE), which combines all steps from rescue to binding in a single microwell. To test this concept, wells were precoated with different antibodies, which allowed newly formed phage particles to be captured directly in situ during overnight rescue. Following 6 washing steps, the retained phages could be easily detected in an enzyme-linked immunosorbent assay (ELISA), thus eliminating the need for purification or concentration of the viral particles. As a consequence, RISE enables a rapid characterization of phage-displayed proteins. In addition, this method allowed for the selective enrichment of phages displaying a hemagglutinin (HA) epitope tag, spiked in a 10(4)-fold excess of wild-type background. Because the combination of phage rescue, selection, or evaluation in a single microwell is amenable to automation, RISE may boost the high-throughput screening of smaller sized phage display libraries. PMID- 15799955 TI - Comparison of 3 AT1 receptor binding assays: filtration assay, ScreenReady Target, and WGA Flashplate. AB - In this article, the study of 3 different angiotensin II type 1 (AT(1)) receptor binding assays in terms of reproducibility, robustness, and feasibility for high throughput screening (HTS) is described. The following methods were used: a nonhomogeneous filtration assay in a 96-well format using CHO-AT(1) cell membranes and 2 homogeneous assays, which include the commercially available ScreenReady Target for the AT(1) receptor and the wheat germ agglutinin (WGA) Flashplate, which was coated "in-house" with the CHO-AT(1) cell membranes. Receptors were labeled with [(125)I]-Sar(1)-Ile(8)-angiotensin II, and radioligand binding was displaced using the antagonist losartan and the natural agonist angiotensin II. Reproducible K(d), B(max), and K(i) values and good total binding/nonspecific binding (TB/NSB) ratios were obtained with both the ScreenReady Targets and the filtration assay, whereas the WGA Flashplates showed unacceptably high nonspecific binding and high variation when applied as a homogeneous assay. However, when applied as a heterogeneous assay (i.e., when a wash step at the end of the assay is included), the results were significantly better. Interestingly, ligand affinities were consistently lower in Flashplate based assays than in the filtration assay. This may be due to the immobilization of the receptors onto the solid surface of the plate, affecting their conformation. In terms of reproducibility, robustness, and feasibility for HTS, the authors conclude that the ScreenReady Target plates are most suitable for AT(1) receptor binding screening. PMID- 15799956 TI - Use of Caenorhabditis elegans G{alpha}q chimeras to detect G-protein-coupled receptor signals. AB - G-protein-coupled receptors (GPCRs) activate heterotrimeric G-proteins (G(i)-, G(s)-, G(q)-, or G(12)-like) to generate specific intracellular responses, depending on the receptor/G-protein coupling. The aim was to enable a majority of GPCRs to generate a predetermined output by signaling through a single G-protein supported pathway. The authors focused on calcium responses as the output, then engineered Galpha(q) to promote promiscuous receptor interactions. Starting with a human Galpha(q) containing 5 Galpha(z) residues in the C-terminal receptor recognition domain (hGalpha(q/z5)), they evaluated agonist-stimulated calcium responses for 33 diverse GPCRs (G(i)-, G(s)-, and G(q)-coupled) and found 20 of 33 responders. In parallel, they tested Caenorhabditis elegans Galpha(q) containing 5 or 9 C-terminal Galpha(z) residues (cGalpha(q/z5), cGalpha(q/z9)). Signal detection was enhanced with cGalpha(q/z5) and cGalpha(q/z9) (yielding 25/33 and 26/33 responders, respectively). In a separate study of Galpha(s) coupled receptors, the authors compared hGalpha(q/s5) versus hGalpha(q/s9), cGalpha(q/s9), andcGalphaq/s21 and observed optimal function with cGalpha(q/s9). Cotransfection of an engineered Galpha(q) "cocktail" (cGalpha(q/z5) plus cGalpha(q/s9)) provided a powerful and efficient screening platform. When the chimeras included N-terminal myristoylation sites (to promote membrane localization), calcium responses were sustained or improved, depending on the receptor. This approach toward a "universal functional assay" is particularly useful for orphan GPCRs whose signaling pathways are unknown. PMID- 15799957 TI - Homogeneous, bioluminescent protease assays: caspase-3 as a model. AB - Using caspase-3 as a model, the authors have developed a strategy for highly sensitive, homogeneous protease assays suitable for high-throughput, automated applications. The assay uses peptide-conjugated aminoluciferin as the protease substrate and a firefly luciferase that has been molecularly evolved for increased stability. By combining the proluminescent caspase-3 substrate, Z-DEVD aminoluciferin, with a stabilized luciferase in a homogeneous format, the authors developed an assay that is significantly faster and more sensitive than fluorescent caspase-3 assays. The assay has a single-step format, in which protease cleavage of the substrate and luciferase oxidation of the aminoluciferin occurs simultaneously. Because these processes are coupled, they rapidly achieve steady state to maintain stable luminescence for several hours. Maximum sensitivity is attained when this steady state occurs; consequently, this coupled enzyme system results in a very rapid assay. The homogeneous format inherently removes trace contamination by free aminoluciferin, resulting in extremely low background and yielding exceptionally high signal-to-noise ratios and excellent Z' factors. Another advantage of a luminescent format is that it avoids problems of cell autofluorescence or fluorescence interference that can be associated with synthetic chemical and natural product libraries. This bioluminescent, homogeneous format should be widely applicable to other protease assays. PMID- 15799958 TI - Development of a microplate-based scintillation proximity assay for MraY using a modified substrate. AB - MraY is an established target for the discovery of antibacterial agents. The conventional assay for MraY uses radioactive substrate and analysis of products after paper chromatography or butanol extraction. Synthesis of radiolabeled substrate has been done in vitro using purified enzymes or by growing cells on radiolabeled precursors. The authors report a simple and rapid method to chemically radiolabel MraY substrate, UDP-MurNAc-pentapeptide. Specific activity obtained by this method was more than 100 times higher than the conventionally labeled substrate, and yields are high enough to support the requirements of high throughput screening (HTS). The authors have developed a microplate-based homogeneous assay for MraY in which the product is captured on wheat germ agglutinin (WGA) scintillation proximity assay (SPA) beads. The assay was validated by showing inhibition by specific inhibitors of MraY but not by inhibitors of other enzymes of peptidoglycan synthesis. The assay uses wild-type membranes of Escherichia coli, giving it an advantage over recently described assays that need the protein to be overexpressed. In addition, it has an advantage over the high-throughput MraY-MurG coupled assay reported in the literature because it is MraY specific, and therefore hits obtained in this assay do not need further deconvolution. It has potential for use in HTS approaches to find novel inhibitors of MraY. PMID- 15799959 TI - Differential inhibition of inducible T cell cytokine secretion by potent iron chelators. AB - Effector functions and proliferation of T helper (Th) cells are influenced by cytokines in the environment. Th1 cells respond to a synergistic effect of interleukin-12 (IL-12) and interleukin-18 (IL-18) to secrete interferon-gamma (IFN-gamma). In contrast, Th2 cells respond to interleukin-4 (IL-4) to secrete IL 4, interleukin-13 (IL-13), interleukin-5 (IL-5), and interleukin-10 (IL-10). The authors were interested in identifying nonpeptide inhibitors of the Th1 response selective for the IL-12/IL-18-mediated secretion of IFN-gamma while leaving the IL-4-mediated Th2 cytokine secretion relatively intact. The authors established a screening protocol using human peripheral blood mononuclear cells (PBMCs) and identified the hydrazino anthranilate compound 1 as a potent inhibitor of IL 12/IL-18-mediated IFN-gamma secretion from CD3(+) cells with an IC(50) around 200 nM. The inhibitor was specific because it had virtually no effect on IL-4 mediated IL-13 release from the same population of cells. Further work established that compound 1 was a potent intracellular iron chelator that inhibited both IL-12/IL-18- and IL-4-mediated T cell proliferation. Iron chelation affects multiple cellular pathways in T cells. Thus, the IL-12/IL-18 mediated proliferation and IFN-gamma secretion are very sensitive to intracellular iron concentration. However, the IL-4-mediated IL-13 secretion does not correlate with proliferation and is partially resistant to potent iron chelation. PMID- 15799960 TI - Identifying modulators of hERG channel activity using the PatchXpress planar patch clamp. AB - The authors used the PatchXpress 7000A system to measure compound activity at the hERG channel using procedures that mimicked the "gold-standard" conventional whole-cell patch clamp. A set of 70 compounds, including hERG antagonists with potencies spanning 3 orders of magnitude, were tested on hERG302-HEK cells using protocols aimed at either identifying compound activity at a single concentration or obtaining compound potency from a cumulative concentration dependence paradigm. After exposure to compounds and subsequent washout of the wells to determine reversibility of the block, blockade by a reference compound served as a quality control. Electrical parameters and voltage dependence were similar to those obtained using a conventional whole-cell patch clamp. Rank order of compound potency was also comparable to that determined by conventional methods. One exception was flunarizine, a particularly lipophilic compound. The PatchXpress accurately identified the activity of 29 moderately potent antagonists, which only weakly displace radiolabeled astemizole and are false negatives in the binding assay. Finally, no false hits were observed from a collection of relatively inactive compounds. High-quality data acquisition by PatchXpress should help accelerate secondary screening for ion channel modulators and the drug discovery process. PMID- 15799962 TI - Proteins in human brain cortex are modified by oxidation, glycoxidation, and lipoxidation. Effects of Alzheimer disease and identification of lipoxidation targets. AB - Diverse oxidative pathways, such as direct oxidation of amino acids, glycoxidation, and lipoxidation could contribute to Alzheimer disease pathogenesis. A global survey for the amount of structurally characterized probes for these reactions is lacking and could overcome the lack of specificity derived from measurement of 2,4-dinitrophenylhydrazine reactive carbonyls. Consequently we analyzed (i) the presence and concentrations of glutamic and aminoadipic semialdehydes, N(epsilon)-(carboxymethyl)-lysine, N(epsilon)-(carboxyethyl) lysine, and N(epsilon)-(malondialdehyde)-lysine by means of gas chromatography/mass spectrometry, (ii) the biological response through expression of the receptor for advanced glycation end products, (iii) the fatty acid composition in brain samples from Alzheimer disease patients and age-matched controls, and (iv) the targets of N(epsilon)-(malondialdehyde)-lysine formation in brain cortex by proteomic techniques. Alzheimer disease was associated with significant, although heterogeneous, increases in the concentrations of all evaluated markers. Alzheimer disease samples presented increases in expression of the receptor for advanced glycation end products with high molecular heterogeneity. Samples from Alzheimer disease patients also showed content of docosahexaenoic acid, which increased lipid peroxidizability. In accordance, N(epsilon)-(malondialdehyde)-lysine formation targeted important proteins for both glial and neuronal homeostasis such as neurofilament L, alpha-tubulin, glial fibrillary acidic protein, ubiquinol-cytochrome c reductase complex protein I, and the beta chain of ATP synthase. These data support an important role for lipid peroxidation-derived protein modifications in Alzheimer disease pathogenesis. PMID- 15799963 TI - Unsaturated fatty acids induce cytotoxic aggregate formation of amyotrophic lateral sclerosis-linked superoxide dismutase 1 mutants. AB - Formation of misfolded protein aggregates is a remarkable hallmark of various neurodegenerative diseases including Alzheimer disease, Parkinson disease, Huntington disease, prion encephalopathies, and amyotrophic lateral sclerosis (ALS). Superoxide dismutase 1 (SOD1) immunoreactive inclusions have been found in the spinal cord of ALS animal models and patients, implicating the close involvement of SOD1 aggregates in ALS pathogenesis. Here we examined the molecular mechanism of aggregate formation of ALS-related SOD1 mutants in vitro. We found that long-chain unsaturated fatty acids (FAs) promoted aggregate formation of SOD1 mutants in both dose- and time-dependent manners. Metal deficient SOD1s, wild-type, and mutants were highly oligomerized compared with holo-SOD1s by incubation in the presence of unsaturated FAs. Oligomerization of SOD1 is closely associated with its structural instability. Heat-treated holo SOD1 mutants were readily oligomerized by the addition of unsaturated FAs, whereas wild-type SOD1 was not. The monounsaturated FA, oleic acid, directly bound to SOD1 and was characterized by a solid-phase FA binding assay using oleate-Sepharose. The FA binding characteristics were closely correlated with the oligomerization propensity of SOD1 proteins, which indicates that FA binding may change SOD1 conformation in a way that favors the formation of aggregates. High molecular mass aggregates of SOD1 induced by FAs have a granular morphology and show significant cytotoxicity. These findings suggest that SOD1 mutants gain FA binding abilities based on their structural instability and form cytotoxic granular aggregates. PMID- 15799964 TI - Wallerian degeneration involves Rho/Rho-kinase signaling. AB - Local axon degeneration is a common pathological feature of many neurodegenerative diseases, whereas the underlying molecular mechanisms are largely unknown. In this study, we used the degeneration of transected axons, termed "Wallerian degeneration," as a model to examine the possible involvement of Rho. Nogo-66, a myelin-derived inhibitor of axon regeneration, significantly accelerated axon degeneration of the dorsal root ganglion explant in vitro, whereas inhibiting Rho-kinase activity abolished the effect. Rho activation was observed in the distal part of the injured axons after spinal cord injury. We demonstrate that degeneration of the injured cortico-spinal axons was significantly retarded by a Rho-kinase inhibitor in vivo. Our findings suggest that inhibiting the signaling pathway may retard axon degeneration in pathological conditions. PMID- 15799965 TI - A family of K+ channel ancillary subunits regulate taste sensitivity in Caenorhabditis elegans. AB - We have identified a family of ancillary subunits of K(+) channels in Caenorhabditis elegans. MPS-1 and its related members MPS-2, MPS-3, and MPS-4 are detected in the nervous system of the nematode. Electrophysiological analysis in ASE neurons and mammalian cells and epigenetic inactivation by double-stranded RNA interference (RNAi) in vivo show that each MPS can associate with and functionally endow the voltage-gated K(+) channel KVS-1. In the chemosensory neuron ADF, three different MPS subunits combine with KVS-1 to form both binary (MPS-1.KVS-1) and ternary (MPS-2.MPS-3.KVS-1) complexes. RNAi of mps-2, mps-3, or both, enhance the taste of the animal for sodium without altering the susceptibility to other attractants. When sodium is introduced in the test plate as background or as antagonist attractant, the nematode loses the ability to recognize a second attractant. Thus, it appears that the chemosensory apparatus of C. elegans uses sensory thresholds and that a voltage-gated K(+) channel is specifically required for this mechanism. PMID- 15799967 TI - Taurine chloramine, an oxidant derived from neutrophils, induces apoptosis in human B lymphoma cells through mitochondrial damage. AB - Taurine chloramine (TN-Cl) is one of the most abundant compounds generated by activated neutrophils. In contrast to HOCl, which causes necrosis, TN-Cl is a potent inducer of apoptosis in tumor cells. Here we show that the apoptosis induced by TN-Cl in human B lymphoma cells is dependent upon oxidant-mediated mitochondrial damage, a decrease in mitochondrial membrane potential, and caspase 9 activation. Further, we show that TN-Cl is taken up into the cells and is concentrated in the mitochondria, where it induces opening of the permeability transition pore and mitochondrial swelling. Identical activity is seen upon treatment of isolated mitochondria with TN-Cl and is blocked by the permeability transition pore inhibitors bongkrekic acid and cyclosporin A, as well as by the sulfhydryl-reducing agent tris(2-carboxyethyl)-phosphine. The data suggest that TN-Cl causes apoptosis through direct damage to the mitochondria. PMID- 15799966 TI - COMMD proteins, a novel family of structural and functional homologs of MURR1. AB - MURR1 is a multifunctional protein that inhibits nuclear factor kappaB (NF kappaB), a transcription factor with pleiotropic functions affecting innate and adaptive immunity, apoptosis, cell cycle regulation, and oncogenesis. Here we report the discovery of a new family of proteins with homology to MURR1. These proteins form multimeric complexes and were identified in a biochemical screen for MURR1-associated factors. The family is defined by the presence of a conserved and unique motif termed the COMM (copper metabolism gene MURR1) domain, which functions as an interface for protein-protein interactions. Like MURR1, several of these factors also associate with and inhibit NF-kappaB. The proteins designated as COMMD or COMM domain containing 1-10 are extensively conserved in multicellular eukaryotic organisms and define a novel family of structural and functional homologs of MURR1. The prototype of this family, MURR1/COMMD1, suppresses NF-kappaB not by affecting nuclear translocation or binding of NF kappaB to cognate motifs; rather, it functions in the nucleus by affecting the association of NF-kappaB with chromatin. PMID- 15799968 TI - Concerted auto-regulation in yeast endosomal t-SNAREs. AB - In yeast, the assembly of the target (t)-SNAREs [Tlg2p/Tlg1p,Vti1p] and [Pep12p/Tlg1p,Vti1p] with the vesicular (v)-SNARE Snc2p promotes endocytic fusion. Here, selected mutations and truncations of SNARE proteins were tested in an in vitro fusion assay to identify potential regulatory regions in these proteins, and two distinct regions were found. The first is represented by the combined effect of the three t-SNARE N-terminal regions and the second is located within the Tlg1p SNARE motif. These internal controls provide a potential mechanism to enable SNARE-dependent fusion to be regulated. PMID- 15799969 TI - Nuclear targeting of transforming growth factor-beta-activated Smad complexes. AB - Upon stimulation by the transforming growth factor beta (TGF-beta), Smad2 and Smad3 are phosphorylated at their C termini and assemble into stable heteromeric complexes with Smad4. These complexes are the functional entities that translocate into the nucleus and regulate the expression of TGF-beta target genes. Here we report that the TGF-beta-activated phospho-Smad3/Smad4 complex utilizes an importin-independent mechanism for nuclear import and engages different nucleoporins for nuclear import compared with the monomeric Smad4. Within the heteromeric complex, phospho-Smad3 appears to dominate over Smad4 in the nuclear import process and guides the complex to its nuclear destination. We also demonstrate that the binding of phospho-Smad3 to Smad4 prevents Smad4 from interacting with the nuclear export receptor chromosome region maintenance 1. In this way, TGF-beta signaling suppresses nuclear export of Smad4 by chromosome region maintenance 1 and thereby targets Smad4 into the nucleus. Indeed tumorigenic mutations in Smad4 that affect its interaction with Smad2 or Smad3 impair nuclear accumulation of Smad4 in response to TGF-beta. PMID- 15799970 TI - The unfolded protein response modulates toxicity of the expanded glutamine androgen receptor. AB - Kennedy disease, a degenerative disorder caused by an expanded glutamine tract, is mediated by misfolding of the mutant androgen receptor (AR) protein, a process that may disrupt proteasome function. We hypothesized that this might lead to endoplasmic reticulum (ER) stress and induction of the unfolded protein response (UPR), a complex physiologic pathway that regulates cell survival. To test this hypothesis, we used aminoterminal fragments of wild type (AR16Q) or mutant (AR112Q) AR that triggered glutamine length-dependent cell death and activated an ER stress-inducible promoter. To evaluate the role of the UPR, we examined the contributions of three proximal sensors of ER stress: activating transcription factor 6 (ATF6), inositol requiring 1 (IRE1), and PKR-like endoplasmic reticulum kinase (PERK). AR112Q toxicity was significantly increased by a dominant negative ATF6 mutant and significantly decreased by a constitutively active ATF6 mutant, indicating that ATF6 promoted cell survival. In contrast, co-transfection with three separate IRE1alpha dominant negative mutants failed to alter glutamine length-dependent toxicity, suggesting that this arm of the UPR did not significantly affect AR112Q induced cell death. Activation of PERK, an ER transmembrane protein that functions as the third proximal UPR sensor, promoted glutamine length-dependent toxicity. Although nuclear localization sequence- and nuclear export sequence-targeted proteins both activated the UPR, this pathway more potently influenced toxicity when proteins were targeted to the cytoplasm. Taken together, our data demonstrate that the UPR is activated in cells expressing long glutamine tracts and that this pathway modulates polyglutamine toxicity. PMID- 15799971 TI - Identification of WNK1 as a substrate of Akt/protein kinase B and a negative regulator of insulin-stimulated mitogenesis in 3T3-L1 cells. AB - Insulin signaling through protein kinase Akt/protein kinase B (PKB), a downstream element of the phosphatidylinositol 3-kinase (PI3K) pathway, regulates diverse cellular functions including metabolic pathways, apoptosis, mitogenesis, and membrane trafficking. To identify Akt/PKB substrates that mediate these effects, we used antibodies that recognize phosphopeptide sites containing the Akt/PKB substrate motif (RXRXX(p)S/T) to immunoprecipitate proteins from insulin stimulated adipocytes. Tryptic peptides from a 250-kDa immunoprecipitated protein were identified as the protein kinase WNK1 (with no lysine) by matrix-assisted laser desorption ionization time-of-flight mass spectrometry, consistent with a recent report that WNK1 is phosphorylated on Thr60 in response to insulin-like growth factor I. Insulin treatment of 3T3-L1 adipocytes stimulated WNK1 phosphorylation, as detected by immunoprecipitation with antibody against WNK1 followed by immunoblotting with the anti-phosphoAkt substrate antibody. WNK1 phosphorylation induced by insulin was unaffected by rapamycin, an inhibitor of p70 S6 kinase pathway but abolished by the PI3K inhibitor wortmannin. RNA interference-directed depletion of Akt1/PKB alpha and Akt2/PKB beta attenuated insulin-stimulated WNK1 phosphorylation, but depletion of protein kinase C lambda did not. Whereas small interfering RNA-induced loss of WNK1 protein did not significantly affect insulin-stimulated glucose transport in 3T3-L1 adipocytes, it significantly enhanced insulin-stimulated thymidine incorporation by about 2 fold. Furthermore, depletion of WNK1 promoted serum-stimulated cell proliferation of 3T3-L1 preadipocytes, as evidenced by a 36% increase in cell number after 48 h in culture. These data suggest that WNK1 is a physiologically relevant target of insulin signaling through PI3K and Akt/PKB and functions as a negative regulator of insulin-stimulated mitogenesis. PMID- 15799972 TI - Membrane depolarization induces the undulating phosphorylation/dephosphorylation of glycogen synthase kinase 3beta, and this dephosphorylation involves protein phosphatases 2A and 2B in SH-SY5Y human neuroblastoma cells. AB - Changes in plasma membrane electrical potential evoke signals that regulate the expressions of various genes in the nervous system. However, the role of glycogen synthase kinase 3beta (GSK-3beta) in this process has not been elucidated. Thus, this study was performed to examine whether membrane depolarization can regulate the phosphorylation of GSK-3beta and to identify the molecular mechanisms involved in this regulation. The depolarization by treating with 100 mm KCl for 5 min resulted in the undulating phosphorylation of GSK-3beta at Ser-9 in SH-SY5Y human neuroblastoma cells, in H19 -7/IGF-IR rat embryonic hippocampal cells, and in PC12 rat pheochromocytoma cells, but not in A172 human glioblastoma cells. Cellular beta-catenin contents showed a temporal pattern similar to that of the Ser-9 phosphorylation of GSK-3beta. Treatment with wortmannin or calphostin C or the expression of dominant negative Akt inhibited phosphorylation of GSK-3beta at Ser-9 following the KCl-induced depolarization of SH-SY5Y cells. Moreover, pretreatment with okadaic acid or cyclosporin A blocked the dephosphorylation of GSK-3beta at Ser-9 at 0, 15, and 30 min after KCl-induced depolarization, and the activity of protein phosphatases (PP) 2A and 2B increased at these times. Treatment with nifedipine or calcium-free medium inhibited GSK-3beta dephosphorylation following membrane depolarization, and the amounts of co immunoprecipitated GSK-3beta and PP2A changed in parallel with GSK-3beta dephosphorylation. Our study demonstrated that KCl-induced depolarization caused undulating GSK-3beta phosphorylation/dephosphorylation, which was regulated for the most part by phosphatidylinositol 3-kinase and Akt (phosphorylation) and PP2A and PP2B (dephosphorylation), respectively. PMID- 15799973 TI - Age-dependent motor deficits and dopaminergic dysfunction in DJ-1 null mice. AB - Mutations in the DJ-1 gene were recently identified in an autosomal recessive form of early-onset familial Parkinson disease. Structural biology, biochemistry, and cell biology studies have suggested potential functions of DJ-1 in oxidative stress, protein folding, and degradation pathways. However, animal models are needed to determine whether and how loss of DJ-1 function leads to Parkinson disease. We have generated DJ-1 null mice with a mutation that resembles the large deletion mutation reported in patients. Our behavioral analyses indicated that DJ-1 deficiency led to age-dependent and task-dependent motoric behavioral deficits that are detectable by 5 months of age. Unbiased stereological studies did not find obvious dopamine neuron loss in 6-month- and 11-month-old mice. Neurochemical examination revealed significant changes in striatal dopaminergic function consisting of increased dopamine reuptake rates and elevated tissue dopamine content. These data represent the in vivo evidence that loss of DJ-1 function alters nigrostriatal dopaminergic function and produces motor deficits. PMID- 15799974 TI - Domain interactions of the mannose 6-phosphate/insulin-like growth factor II receptor. AB - The mannose 6-phosphate/insulin-like growth factor II receptor (M6P/IGF2R) forms oligomeric structures important for optimal function in binding and internalization of Man-6-P-bearing extracellular ligands as well as lysosomal biogenesis and growth regulation. However, neither the mechanism of inter receptor interaction nor the dimerization domain has yet been identified. We hypothesized that areas near the ligand binding domains of the receptor would contribute preferentially to oligomerization. Two panels of minireceptors were constructed that involved truncations of either the N- or C-terminal regions of the M6P/IGF2R encompassing deletions of various ligand binding domains. alpha FLAG or alpha-Myc-based immunoprecipitation assays showed that all of the minireceptors tested were able to associate with a full-length, Myc-tagged M6P/IGF2R (WT-M). In the alpha-FLAG but not alpha-Myc immunoprecipitation assays, the degree of association of a series of C-terminally truncated minireceptors with WT-M showed a positive trend with length of the minireceptor. In contrast, length did not seem to affect the association of the N-terminally truncated minireceptors with WT-M, except that the 12th extracytoplasmic repeat appeared exceptionally important in dimerization in the alpha-FLAG assays. The presence of mutations in the ligand-binding sites of the minireceptors had no effect on their ability to associate with WT-M. Thus, association within the heterodimers was not dependent on the presence of functional ligand binding domains. Heterodimers formed between WT-M and the minireceptors demonstrated high affinity IGF-II and Man-6-P-ligand binding, suggesting a functional association. We conclude that there is no finite M6P/IGF2R dimerization domain, but rather that interactions between dimer partners occur all along the extracytoplasmic region of the receptor. PMID- 15799975 TI - Acid-triggered membrane insertion of Pseudomonas exotoxin A involves an original mechanism based on pH-regulated tryptophan exposure. AB - Exposure to low endosomal pH during internalization of Pseudomonas exotoxin A (PE) triggers membrane insertion of its translocation domain. This process is a prerequisite for PE translocation to the cytosol where it inactivates protein synthesis. Although hydrophobic helices enable membrane insertion of related bacterial toxins such as diphtheria toxin, the PE translocation domain is devoid of hydrophobic stretches and the structural features triggering acid-induced membrane insertion of PE are not known. Here we have identified a molecular device that enables PE membrane insertion. This process is promoted by exposure of a key tryptophan residue. At neutral pH, this Trp is buried in a hydrophobic pocket closed by the smallest alpha-helix of the translocation domain. Upon acidification, protonation of the Asp that is the N-cap residue of the helix leads to its destabilization, enabling Trp side chain insertion into the endosome membrane. This tryptophan-based membrane insertion system is surprisingly similar to the membrane-anchoring mechanism of human annexin-V and could be used by other proteins as well. PMID- 15799976 TI - Identification of a novel pharmacophore for peptide toxins interacting with K+ channels. AB - KappaM-conotoxin RIIIK blocks TSha1 K+ channels from trout with high affinity by interacting with the ion channel pore. As opposed to many other peptides targeting K+ channels, kappaM-RIIIK does not possess a functional dyad. In this study we combine thermodynamic mutant cycle analysis and docking calculations to derive the binding mode of kappaM-conotoxin RIIIK to the TSha1 channel. The final model reveals a novel pharmacophore, where no positively charged side chain occludes the channel pore. Instead the positive-charged residues of the toxin form a basic ring; kappaM-RIIIK is anchored to the K+ channel via electrostatic interactions of this basic ring with the loop and pore helix residues of the channel. The channel amino acid Glu-354 is likely to be a fundamental determinant of the selectivity of kappaM-RIIIK for the TSha1 channel. The Cgamma-OH of Hyp-15 is in contact with the carbonyls of the selectivity filter, disturbing the charge distribution pattern necessary for the coordination of K+ ions. This novel, experimentally based pharmacophore model proves the existence of diverse binding modes of peptidic toxins to K+ channels and underlines the role of intermolecular electrostatic interactions involving channel loop side chains in determining the selectivity of toxins for specific K+ channel types. PMID- 15799977 TI - Portal fibroblasts regulate the proliferation of bile duct epithelia via expression of NTPDase2. AB - Bile duct epithelia are the target of a number of "cholangiopathies" characterized by disordered bile ductular proliferation. Although mechanisms for bile ductular proliferation are unknown, recent evidence suggests that extracellular nucleotides regulate cell proliferation via activation of P2Y receptors. Portal fibroblasts may regulate bile duct epithelial P2Y receptors via expression of the ecto-nucleotidase NTPDase2. Thus, we tested the hypothesis that portal fibroblasts regulate bile duct epithelial proliferation via expression of NTPDase2. We generated a novel co-culture model of Mz-ChA-1 human cholangiocarcinoma cells and primary portal fibroblasts. Cell proliferation was measured by bromodeoxyuridine uptake. NTPDase2 expression was assessed by immunofluorescence and quantitative real-time reverse transcription PCR. NTPDase2 expression in portal fibroblasts was blocked using short interfering RNA. NTPDase2 overexpression in portal myofibroblasts isolated from bile duct-ligated rats was achieved by cDNA transfection. Co-culture of Mz-ChA-1 cells with portal fibroblasts decreased their proliferation to 26% of control. Similar decreases in Mz-ChA-1 proliferation were induced by the soluble ecto-nucleotidase apyrase and the P2 receptor inhibitor suramin. The proliferation of Mz-ChA-1 cells returned to baseline when NTPDase2 expression in portal fibroblasts was inhibited using NTPDase2-specific short interfering RNA. Untransfected portal myofibroblasts lacking NTPDase2 had no effect on Mz-ChA-1 proliferation, yet portal myofibroblasts transfected with NTPDase2 cDNA inhibited Mz-ChA-1 proliferation. We conclude that portal fibroblasts inhibit bile ductular proliferation via expression of NTPDase2 and blockade of P2Y activation. Loss of NTPDase2 may mediate the bile ductular proliferation typical of obstructive cholestasis. This novel cross-talk signaling pathway may mediate pathologic alterations in bile ductular proliferation in other cholangiopathic conditions. PMID- 15799978 TI - Characterization of the functional insulin binding epitopes of the full-length insulin receptor. AB - Mutational analyses of the secreted recombinant insulin receptor extracellular domain have identified a ligand binding site composed of residues located in the L1 domain (amino acids 1-470) and at the C terminus of the alpha subunit (amino acids 705-715). To evaluate the physiological significance of this ligand binding site, we have transiently expressed cDNAs encoding full-length receptors with alanine mutations of the residues forming the functional epitopes of this binding site and determined their insulin binding properties. Insulin bound to wild-type receptors with complex kinetics, which were fitted to a two-component sequential model; the Kd of the high affinity component was 0.03 nM and that of the low affinity component was 0.4 nM. Mutations of Arg14, Phe64, Phe705, Glu706, Tyr708, Asn711, and Val715 inactivated the receptor. Alanine mutation of Asn15 resulted in a 20-fold decrease in affinity, whereas mutations of Asp12, Gln34, Leu36, Leu37, Leu87, Phe89, Tyr91, Lys121, Leu709, and Phe714 all resulted in 4-10-fold decreases. When the effects of the mutations were compared with those of the same mutations of the secreted recombinant receptor, significant differences were observed for Asn15, Leu37, Asp707, Leu709, Tyr708, Asn711, Phe714, and Val715, suggesting that the molecular basis for the interaction of each form of the receptor with insulin differs. We also examined the effects of alanine mutations of Asn15, Gln34, and Phe89 on insulin-induced receptor autophosphorylation. They had no effect on the maximal response to insulin but produced an increase in the EC50 commensurate with their effect on the affinity of the receptor for insulin. PMID- 15799979 TI - Future development of nursing home quality indicators. AB - Nursing home quality indicators have been developed over the past 10 years to quantify nursing home quality and to draw systematic comparisons between facilities. Although these indicators have been applied widely for nursing home regulation, quality improvement, and public reporting, researchers and stakeholders have raised concerns about their accuracy and usefulness. We critically evaluate nursing home quality indicators from the standpoint of theory, measurement, and application, and we recommend strategies to make the indicators more valuable as quality assessment tools. We recommend that (a) more comprehensive quality indicators should be developed in conjunction with the new Minimum Data Set 3.0; (b) the validity and reliability of the indicators should be evaluated thoroughly with respect to both measurement and application; (c) statistical criteria should be incorporated explicitly into quality indicator scoring and outlier targeting; (d) the dimensionality and theoretical structure of the quality indicators should be carefully examined; (e) risk adjustment methods should be refined and broadened; and (f) quality indicator reporting systems should be strengthened and tailored to stakeholders' needs. PMID- 15799980 TI - Characteristics predicting nursing home admission in the program of all-inclusive care for elderly people. AB - PURPOSE: This study determined overall risk and predictors of long-term nursing home admission within the Program of All-Inclusive Care for the Elderly (PACE). DESIGN AND METHODS: DataPACE records for 4,646 participants aged 55 years or older who were enrolled in 12 Medicare- and Medicaid-capitated PACE programs during the period from June 1, 1990, to June 30, 1998, were obtained. Participants were enrolled for at least 30 days and had baseline evaluations within 30 days of enrollment. Cox proportional hazard models predicting an outcome of nursing home admission of 30 days or longer were estimated. RESULTS: The cumulative risk of admission to nursing homes for 30 days or longer was 14.9% within 3 years. Individuals enrolled from a nursing home were at very high risk for future admission, with a relative risk of 5.20 when compared with those living alone. Among individuals enrolled in PACE from the community, age, instrumental activity of daily living dependence, and bowel incontinence were predictive of subsequent nursing home admission. Asians and Blacks had a lower risk of institutionalization than Whites. However, other characteristics were not independently predictive of institutionalization, namely poor cognitive status, number of chronic conditions, activity of daily living deficits, urinary incontinence, several behavioral disturbances, and duration of program operation. Before adjusting for other variables, there was substantial site variability in risk of nursing home admission; this decreased considerably after other characteristics were adjusted for. IMPLICATIONS: Despite the fact that 100% of the PACE participants were nursing home certifiable, the risk of being admitted to a nursing home long term following enrollment from the community is low. The presence of some reversible risk factors may have implications for early intervention to reduce risk further, although the effect of these interventions is likely to be modest. Individuals who received long-term care in a nursing home prior to enrollment in PACE remain at high risk of readmission, despite the availability of comprehensive services. PMID- 15799981 TI - Effect of a consumer-directed voucher and a disease-management-health-promotion nurse intervention on home care use. AB - PURPOSE: We describe the impact of two interventions, a consumer-directed voucher for in-home supportive services and a chronic disease self-management-health promotion nurse intervention, on the probability of use of two types of home care skilled home health care and personal assistance services-received by functionally impaired Medicare beneficiaries. DESIGN AND METHODS: The Medicare Primary and Consumer-Directed Care Demonstration was a randomized controlled trial in 19 counties in New York, West Virginia, and Ohio with four groups: disease-management-health-promotion nurse, consumer-directed voucher, combination (nurse plus voucher), and control. We estimated a bivariate probit model for the 1,394 individuals aged 65 or older who had no private long-term-care insurance. RESULTS: Whereas the nurse intervention alone had no effect on the probability of using either type of home care, the voucher alone increased the probability of personal assistance services use by 13% (p =.002) but not that of skilled home health care use. The combination of the two interventions increased the probability of personal assistance services use by 18% (p <.001). IMPLICATIONS: The implementation of disease-management-health-promotion nurse interventions should not lead to a greater probability of skilled home health care or personal assistance services use, whereas the provision of consumer-directed vouchers should result in an increased probability of personal assistance services use, as intended. PMID- 15799982 TI - Early community-based service utilization and its effects on institutionalization in dementia caregiving. AB - PURPOSE: The present study attempts to determine whether utilizing community based long-term-care services early in the dementia caregiving career delays time to nursing home placement (adjusting for severity of dementia). DESIGN AND METHODS: With a reliance on data from 4,761 dementia caregivers recruited from eight catchment areas in the United States and followed over a 3-year period, a Cox proportional hazards model was conducted that considered key components of the stress process (e.g., context of care, primary objective and subjective stressors, and resources), duration, and community-based long-term-care use. RESULTS: An analysis of interaction terms in the Cox regression model found that those individuals who utilized in-home help services earlier in their dementia caregiving careers were more likely to delay institutionalization. IMPLICATIONS: The findings suggest the practical importance and cost-effectiveness implications of early community-based service use, and they emphasize the role of timing when one is conceptualizing the proliferation of stress in the dementia caregiving career. PMID- 15799983 TI - Turnover begets turnover. AB - PURPOSE: This study examined the association between turnover of caregivers and turnover of nursing home top management. The top managers examined were administrators and directors of nursing, and the caregivers examined were registered nurses, licensed practical nurses, and nurse aides. DESIGN AND METHODS: The data came from a survey of 419 nursing facilities and the Online Survey, Certification, and Reporting system. Multinomial logistic regression analyses were used to examine the association between turnover of nursing home top management and turnover of caregivers. RESULTS: A 10% increase in top management turnover is associated (p <.05) with a 21% increase in the odds that a facility will have a high turnover rate of nurse aides and is associated (p <.05) with an 8% decrease in the odds that a facility will have a low turnover rate of nurse aides. A 10% increase in top management turnover is associated (p <.1) with a 30% increase in the odds that a facility will have a high turnover rate for registered and licensed practical nurses. IMPLICATIONS: This study provides preliminary evidence that the turnover of top managers may have an important influence on caregiver turnover in nursing homes. PMID- 15799984 TI - Predictors of organizational commitment among staff in assisted living. AB - PURPOSE: This study examines the role of organizational culture, job satisfaction, and sociodemographic characteristics as predictors of organizational commitment among staff in assisted living. It is particularly important to examine organizational commitment, because of its close links to staff turnover. DESIGN AND METHODS: Data were collected from 317 staff members in 61 facilities, using self-administered questionnaires. The facilities were selected from licensed assisted living programs and were stratified into small, traditional, and new-model homes. Staff questionnaires were distributed by a researcher during 1-day visits to each facility. Organizational commitment was measured by the extent of staff identification, involvement, and loyalty to the organization. RESULTS: Organizational culture, job satisfaction, and education were strong predictors of commitment, together explaining 58% of the total variance in the dependent variable. Higher levels of organizational commitment were associated with more favorable staff perceptions of organizational culture and greater job satisfaction. In addition, more educated staff members tended to report higher levels of organizational commitment. Other than education, sociodemographic characteristics failed to account for a significant amount of variance in organizational commitment. IMPLICATIONS: Because job satisfaction and organizational culture were strong predictors of commitment, interventions aimed at increasing job satisfaction and creating an organizational culture that values and respects staff members could be most effective in producing higher levels of organizational commitment. PMID- 15799985 TI - Elder disability as an explanation for racial differences in informal home care. AB - PURPOSE: Adjusting for sociodemographic characteristics and disability levels, this study examines whether differences exist in the structure and function of community-dwelling Black and White frail elders' informal care networks. DESIGN AND METHODS: Data from in-person assessments of Michigan's Home and Community Based Medicaid Waiver applicants were analyzed by using logistic and ordinary least squares regression. The sample consisted of 936 Black and 3,182 White frail elderly persons. RESULTS: When sociodemographic characteristics were controlled for, racial differences were found in all informal care components except out-of home chores. Differences in functional components (amount of care, scope of assistance, and personal care) were largely accounted for by disability, whereas racial differences in the structural components (source of care, living arrangement, and sole caregiver) and in-home chores were not. IMPLICATIONS: Findings suggest that Blacks are not better off than Whites in the receipt of informal care. Although Black elders receive more informal help, this difference is primarily because Blacks are more disabled. This study calls for heightened awareness of disability among low-income Blacks and the potential burden experienced by their caregivers. PMID- 15799986 TI - Change in self-rated health and mortality among community-dwelling disabled older women. AB - PURPOSE: Our study assessed whether change in self-rated health is a stronger predictor of mortality than baseline self-rated health and the most recent self rated health (prior to death or loss to follow-up) among disabled older women. DESIGN AND METHODS: The Women's Health and Aging Study examined disabled older women at baseline and every 6 months for 3 years. During the follow-up period, 253 out of the 905 examined participants died. Cox regression models with time dependent covariates were used. RESULTS: After baseline characteristics were adjusted for, baseline self-rated health was not related to mortality. After covariates at the most recent observation and covariates measured only at baseline were controlled for, the most recent self-rated health was not associated with mortality either. After time-dependent covariates and covariates measured only at baseline were adjusted for, decline in self-rated health was significantly associated with increased mortality. IMPLICATIONS: Change in self rated health is a stronger predictor of mortality than self-rated health at baseline and at the most recent observation. Older women with "fair" health are worse off if they are on a declining health trajectory than if their "fair" health is stable. Family caregivers and clinicians need to closely monitor change in self-rated health among disabled older women. PMID- 15799987 TI - Cognitive impairment as a strong predictor of incident disability in specific ADL IADL tasks among community-dwelling elders: the Azuchi Study. AB - PURPOSE: We examined differential effects of cognitive impairment on each of the activities of daily living (ADL) and instrumental activities of daily living (IADL) tasks. DESIGN AND METHODS: In a 3-year follow-up of community-dwelling elderly persons in Azuchi, Japan, we assessed cognition by using the Hasegawa Dementia Scale. We examined (a) the cross-sectional association between cognitive impairment and functional disability in each ADL-IADL item; (b) cognitive impairment as a risk factor for incident disability in each ADL-IADL item, through logistic regression models; and (c) probabilities of incident loss of ADL IADL abilities or death in 3 years, using multinomial logistic regression models. We also calculated the population attributable risk (PAR%) of cognitive impairment on incident loss of task-specific ADL-IADL abilities. RESULTS: Cross sectionally, the severity of cognitive impairment was associated with disability in each ADL-IADL task, with larger effects shown for ADL items. Longitudinally, minimally or mildly cognitively impaired individuals had a significantly higher risk of losing functional abilities compared with those with intact cognition. The PAR% indicated that cognitive impairment accounts for 11% to 36% of incident disability in ADL-IADL tasks, with the highest PAR% shown for the ability to feed oneself. IMPLICATIONS: Cognitively impaired subjects are heterogeneous; the severity of cognitive impairment has a different impact on incident loss of task specific ADL-IADL abilities, and comorbidities could affect disabilities differently. Consideration of these heterogeneities will enrich future studies on the impact of cognitive impairment on ADL-IADL abilities. PMID- 15799988 TI - Physicians' perspectives on caring for cognitively impaired elders. AB - PURPOSE: This study aims to develop an in-depth understanding of the issues important to primary care physicians in providing care to cognitively impaired elders. DESIGN AND METHODS: In-depth interviews were conducted with 20 primary care physicians. Text coded as "cognitive impairment" was retrieved and analyzed by use of grounded theory analysis techniques. RESULTS: A patient's impaired ability to provide an accurate history and to participate in self-care hindered the usual process of care, often resulting in greater medical uncertainty and feelings of inadequacy and frustration for the physician. Shifting the goal of care from "curing" the patient's illness to "caring" for the patient's quality of life was also problematic. The doctor-patient relationship changed dramatically as others became involved in care, often with attendant ethical dilemmas related to patient autonomy and the locus of decision making. Many physicians described a deep sense of loss and grief as the personhood of patients faded. The increased complexity and prominent social and emotional issues were difficult to manage in the context of the current model of practice. IMPLICATIONS: Profound changes occur in the process of care with cognitively impaired patients. The increased complexity mandates an expanded model of care that addresses the prominent psychosocial and ethical aspects of care as well as the medical ones. PMID- 15799989 TI - Development of the 12-item Expectations Regarding Aging Survey. AB - PURPOSE: This study describes the development of a short version of the Expectations Regarding Aging Survey (ERA-38), a 38-item survey measuring expectations regarding aging. DESIGN AND METHODS: In 1999, surveys containing the ERA-38 were mailed to 588 adults aged > or = 65 years who were recruited through physicians; 429 individuals (73%) returned completed surveys. The mean age of participants was 77 years; 76% were White. In 2001, we surveyed 643 adults aged > or = 65 years recruited at 14 senior centers. The mean age of participants was 78 years; 37% were Latino and 16% were African American. With the 1999 data, we selected items for the shorter version of the ERA-38 by using qualitative criteria and by evaluating the items' factor structure, internal consistency reliability of scales, and correlations with age and self-reported measures of health. Then, using the 2001 data, we evaluated the selected items with confirmatory factor analysis, and we reevaluated the internal consistency reliability and associations of the scales with age and self-reported measures of health. RESULTS: The factor analyses of the ERA-12 on both samples provided support for three 4-item scales (expectations regarding physical health, expectations regarding mental health, and expectations regarding cognitive function), and one global expectations regarding aging scale combining all 12 items. In both samples, internal consistency reliability estimates for all scales exceeded 0.74, and the 12 items together explained over 88% of the variance in the ERA-38 total score. We found comparable associations of the ERA-12 scales with age and self-reported health measures in both samples. IMPLICATIONS: The ERA 12 demonstrated acceptable reliability and validity to estimate expectations regarding aging. PMID- 15799990 TI - Health services utilization between older and younger homeless adults. AB - PURPOSE: Our purpose in the current study was to examine the relationship between health services utilization delivered by means of the Homeless Outreach Medical Services (HOMES) program and health services utilization delivered by means of the Parkland emergency room and inpatient units among a sample of older and younger homeless adults being served by the Parkland Health and Hospital System. DESIGN AND METHODS: The HOMES program uses two 40-ft-long mobile medical units and a fixed-site outpatient clinic to provide primary health care on a walk-in, no-charge basis to individuals who are homeless. We used a quasi-experimental design, and we randomly selected health services utilization data from 293 male (50.4%) and 288 female (49.6%) homeless patients from among the 14,876 homeless patients aged 20-64 years who used HOMES and the Parkland Health and Hospital System between June 1, 1992, and June 30, 1999. The dependent variables were measurements of inpatient and outpatient utilization of psychiatric, substance abuse, and musculoskeletal services by homeless patients. Inpatient and outpatient utilization patterns by age were assessed. Data were analyzed using a split-plot repeated measures multivariate analysis of variance (MANOVA), one-way analysis of variance (ANOVA), and simple linear regression. RESULTS: Older homeless individuals utilized the HOMES more than inpatient services for substance abuse and musculoskeletal conditions, but they utilized inpatient services the most for psychiatric-related conditions. IMPLICATIONS: This level of inpatient utilization is perhaps a result of greater severity or progression of mental illness among older homeless adults. The level of outpatient utilization is probably because the HOMES program mitigates many of the barriers that prevent homeless individuals from obtaining primary medical care. PMID- 15799991 TI - Developing an interdisciplinary innovative Master of Gerontology program. AB - PURPOSE: This study describes the goals, development, and outcomes of an innovative, interdisciplinary, Master of Gerontology department, one of two programs in Israel developed to answer Israel's needs for highly educated researchers and practitioners. DESIGN: Key program components reflecting the program's innovative nature are described: (a) level of development and promotion of knowledge; (b) goals; (c) influence; and (d) patronage. RESULTS: An interdisciplinary faculty and student body both report a high satisfaction level, involvement, and the obtainment of wide knowledge bases in gerontology and geriatrics. The number of candidates for the program increases each year. A new unique program for physicians has evolved, with strong support from the University of Haifa, the Council for Higher Education in Israel, and key service organizations for the elderly population. IMPLICATIONS: Such a program enhances the level of professional leadership in the field of aging, and it expands the cadre of academic faculty. PMID- 15799992 TI - Grandparents raising their grandchildren: a review of the literature and suggestions for practice. AB - An increasingly prevalent family constellation is a home headed by a grandparent who is raising grandchildren. We explore the state of our knowledge about such grandparents with particular attention to its implications for service providers and researchers. In our review we address several key areas: (a) the costs and benefits of raising a grandchild; (b) the heterogeneity of custodial grandparent caregivers; (c) the critical need for social support among custodial grandparents; (d) parenting practices and attitudes among grandparents raising grandchildren; and (e) helping efforts at multiple levels with custodial grandparents. We also discuss directions for research and practice concerning custodial grandparents. PMID- 15799993 TI - Workplace exposures and employment patterns in adult onset asthmatics and diabetics. AB - BACKGROUND: Little is known about the impact of working conditions on employment decisions after the onset of chronic diseases in adult life. AIMS: To determine the effects of job characteristics on employment in patients attending asthma and diabetes clinics. METHODS: Patients aged 16-60 years were interviewed when attending outpatient clinics or undergoing day case surgery. The asthmatics (n=165) and diabetics (n=283) were taking regular prescribed medication for their condition, and were in paid employment at the time of diagnosis. Job titles at diagnosis were rated for exposures with the potential to cause or aggravate asthma or diabetes, and median ratings were used to assess the impact of exposures on the probability of changing the job held at diagnosis. RESULTS: Exposure to irritants or sensitizers at the time of diagnosis was more likely in asthmatics than in the comparison group (odds ratio = 7.09; 95% CI 3.12-16.17); physical activity was unrelated to diagnosis. At interview, on average 8.5 years after diagnosis, more asthmatics were unemployed (44%; mean age 47 years) than diabetics (31%; mean age 49 years) with ill-health being the most common reason given. No relation was found between exposure in the job held at diagnosis and subsequent job change. CONCLUSIONS: No evidence was found that changing jobs was used as a strategy to find more tolerable working conditions. The high proportion of unemployment due to ill-health among adult onset asthmatics deserves further investigation. PMID- 15799994 TI - The decay of blood lead levels in workers suspended under the control of lead at work regulations. AB - AIMS: To study the rate of decline in blood lead levels post-suspension under Control of Lead at Work Regulations (CLAW) and thereby suggest sampling frequencies for follow-up blood lead measurements. METHODS: A retrospective cohort of lead workers with blood lead levels over the current suspension level were identified from blood lead records. Data on their suspension and follow-up blood lead measurements were obtained. RESULTS: Sixteen per cent of the identified cohort did not appear to return to lead work under CLAW. Twenty-seven suspension cases with an initial mean blood lead of 79 microg/dl (3.82 micromol/l) formed the dataset for analysis of decline in blood lead levels. The mean length of time between the blood sample indicating suspension and the first follow-up blood sample was 32 days. The mean length of suspension under CLAW was 61 days. The mean initial rate of blood lead decay was 0.659 microg/dl per day (0.032 micromol/l per day), although with a wide range. The rate of decline in blood lead after suspension was increased by the blood lead level at suspension, but was decreased by increasing past cumulative exposure. CONCLUSIONS: A follow up blood lead sample 1 month after suspension should show a mean decrease between 13 and 26 microg/dl (0.63-1.25 micromol/l), which is substantially greater than that due to analytical 'noise' associated with two sequential measurements (approximately 5 microg/dl). Therefore, a follow-up blood sample taken around 3-4 weeks after suspension would seem practical. A decrease in blood lead of 7-8 mug/dl (0.36 micromol/l) or less in the month after suspension may suggest continuing lead exposure. PMID- 15799995 TI - Validity of the HSE stress tool: an investigation within four organizations by the Corporate Health and Performance Group. AB - AIM: To examine the validity and accuracy of the HSE Management Standards Stress Tool. METHODS: A cross-sectional questionnaire was administered to employees of the occupational health and human resource departments of four of the Corporate Health and Performance Group organizations. The relationship between the six first-pass stress filter questions and self-rated health were examined using the General Health Questionnaire (GHQ12) and the Short Form. The relationship between the filter questions and objective work characteristics such as sickness absence, self-rated performance and job satisfaction was also examined. RESULTS: Two hundred and thirty-five employees participated, giving an overall response rate of 68%. The study sample met the proposed HSE of 85 and 65% pass rates and would not normally progress to further analysis. Almost 40% of the group were above threshold on the GHQ12; 19% rated their work as very or extremely stressful. While the filters were highly specific, they were insensitive and at best detected for less than 40% of those with psychological or work stress. All but one of the filters had less than 50% positive predictive value for work stress and the majority of those identified by the filters did not consider themselves stressed, either overall or by work. Five of the filter questions were significantly associated with self-rated performance, three with absence and two with other objective work indicators. CONCLUSIONS: Further developmental work on these filter questions is indicated. Use of work absence as a measure of the impact or 'cost' of stress may lead to a significant underestimation of the 'true cost' of psychosocial hazards in the workplace. PMID- 15799996 TI - Changing medical criteria and medical severance payments may reduce the rate and costs of ill-health retirement. AB - OBJECTIVES: To identify the core best practice standards in ill-health retirement (IHR) procedures. To investigate whether changing medical criteria and introducing medical severance payments affect the rate and cost of IHR. METHODS: The core standards for best practice in IHR procedures were distilled from the published literature. On 1st April 2000 the study pension scheme altered the IHR medical criteria to define permanent incapacity and introduced medical severance payments for employees with temporary incapacity. Rates and costs of IHR were measured before and after these changes. RESULTS: Following the changes, the annual rate of IHR fell from 8.89 to 2.90 per 1000 members (P < 0.001), the median age at IHR rose from 50 to 55 years (P = 0.01) and pension scheme costs fell by 25 million pounds sterlings per year. CONCLUSIONS: Changing medical criteria and introducing medical severance payments may reduce the rate and costs of ill-health retirement. Target rates of four cases of IHR per 1000 active members per year, and 15% of total retirements, are proposed for schemes serving industries with average health risks. PMID- 15799997 TI - The Arabidopsis gene CAD1 controls programmed cell death in the plant immune system and encodes a protein containing a MACPF domain. AB - To clarify the processes involved in plant immunity, we have isolated and characterized a single recessive Arabidopsis mutant, cad1 (constitutively activated cell death 1), which shows a phenotype that mimics the lesions seen in the hypersensitive response (HR). This mutant shows spontaneously activated expression of pathogenesis-related (PR) genes, and leading to a 32-fold increase in salicylic acid (SA). Inoculation of cad1 mutant plants with Pseudomonas syringae pv tomato DC3000 shows that the cad1 mutation results in the restriction of bacterial growth. Cloning of CAD1 reveals that this gene encodes a protein containing a domain with significant homology to the MACPF (membrane attack complex and perforin) domain of complement components and perforin proteins that are involved in innate immunity in animals. Furthermore, cell death is suppressed in transgenic cad1 plants expressing nahG, which encodes an SA-degrading enzyme. We therefore conclude that the CAD1 protein negatively controls the SA-mediated pathway of programmed cell death in plant immunity. PMID- 15799998 TI - A novel statistical ligand-binding site predictor: application to ATP-binding sites. AB - Structural genomics initiatives are leading to rapid growth in newly determined protein 3D structures, the functional characterization of which may still be inadequate. As an attempt to provide insights into the possible roles of the emerging proteins whose structures are available and/or to complement biochemical research, a variety of computational methods have been developed for the screening and prediction of ligand-binding sites in raw structural data, including statistical pattern classification techniques. In this paper, we report a novel statistical descriptor (the Oriented Shell Model) for protein ligand binding sites, which utilizes the distance and angular position distribution of various structural and physicochemical features present in immediate proximity to the center of a binding site. Using the support vector machine (SVM) as the classifier, our model identified 69% of the ATP-binding sites in whole-protein scanning tests and in eukaryotic proteins the accuracy is particularly high. We propose that this feature extraction and machine learning procedure can screen out ligand-binding-capable protein candidates and can yield valuable biochemical information for individual proteins. PMID- 15799999 TI - The single AmphiTrk receptor highlights increased complexity of neurotrophin signalling in vertebrates and suggests an early role in developing sensory neuroepidermal cells. AB - Neurotrophins (Nt) and their tyrosine kinase Trk receptors play an essential role in the development and maintenance of the complex vertebrate nervous system. Invertebrate genome sequencing projects have suggested that the Nt/Trk system is a vertebrate innovation. We describe the isolation and characterisation of the amphioxus Trk receptor, AmphiTrk. Its ancestral link to vertebrate Trk receptors is supported by phylogenetic analysis and domain characterisation. The genomic structure of AmphiTrk strongly suggests that a ProtoTrk gene emerged by means of exon-shuffling prior to the cephalochordate/vertebrate split. We also examined the physiological response of AmphiTrk to vertebrate neurotrophins, and found that despite 500 million years of divergence, AmphiTrk transduces signals mediated by NGF, BDNF, NT3 and NT4. Markedly, AmphiTrk is able to activate survival and differentiation pathways, but fails to activate the PLCgamma pathway, which is involved in synaptic plasticity in higher vertebrates. AmphiTrk is expressed during amphioxus embryogenesis in sensory neural precursors in the epidermis, which possesses single migratory cells. We propose that the duplication and divergence of the Nt/Trk system, in tandem with recruitment of the PLCgamma pathway, may have provided the genetic basis for a key aspect of vertebrate evolution: the complexity of the nervous system. PMID- 15800000 TI - Fgf10 expression identifies parabronchial smooth muscle cell progenitors and is required for their entry into the smooth muscle cell lineage. AB - Lineage formation in the lung mesenchyme is poorly understood. Using a transgenic mouse line expressing LacZ under the control of Fgf10 regulatory sequences, we show that the pool of Fgf10-positive cells in the distal lung mesenchyme contains progenitors of the parabronchial smooth muscle cells. Fgf10 gene expression is slightly repressed in this transgenic line. This allowed us to create a hypomorphic Fgf10 phenotype by expressing the LacZ transgene in a heterozygous Fgf10 background. Hypomorphic Fgf10 mutant lungs display a decrease in beta galactosidase-positive cells around the bronchial epithelium associated with an accumulation of beta-galactosidase-expressing cells in the distal mesenchyme. This correlates with a marked reduction of alpha smooth muscle actin expression, thereby demonstrating that FGF10 is mostly required for the entry of mesenchymal cells into the parabronchial smooth muscle cell lineage. The failure of exogenous FGF10 to phosphorylate its known downstream targets ERK and AKT in lung mesenchymal cultures strongly suggests that FGF10 acts indirectly on the progenitor population via an epithelial intermediate. We provide support for a role of epithelial BMP4 in mediating the formation of parabronchial smooth muscle cells. PMID- 15800001 TI - The CARD-carrying caspase Dronc is essential for most, but not all, developmental cell death in Drosophila. AB - The initiator caspase Dronc is the only Drosophila caspase that contains a caspase activation and recruitment domain (CARD). Although Dronc has been implicated as an important effector of apoptosis, the genetic function of dronc in normal development is unclear because dronc mutants have not been available. In an EMS mutagenesis screen, we isolated four point mutations in dronc that recessively suppress the eye ablation phenotype caused by eye-specific overexpression of hid. Homozygous mutant dronc animals die during pupal stages; however, at a low frequency we obtained homozygous adult escapers. These escapers have additional cells in the eye and wings that are less transparent and slightly curved down. We determined that this is due to lack of apoptosis. Our analyses of dronc mutant embryos suggest that dronc is essential for most apoptotic cell death during Drosophila development, but they also imply the existence of a dronc independent cell death pathway. We also constructed double mutant flies for dronc and the apoptosis inhibitor diap1. dronc mutants can rescue the ovarian degeneration phenotype caused by diap1 mutations, confirming that dronc acts genetically downstream of diap1. PMID- 15800002 TI - The BLADE ON PETIOLE genes act redundantly to control the growth and development of lateral organs. AB - Developmental processes in multicellular organisms involve an intricate balance between mechanisms that promote cell division activity and growth, and others that promote cell differentiation. Leaf development in Arabidopsis thaliana is controlled by genes like BLADE ON PETIOLE1 (BOP1), which prevent the development of ectopic meristematic activity that leads to the formation of new organs, and JAGGED (JAG), which control the proximodistal development of the leaf by regulating cell-division activity. We have isolated and characterized the BOP1 gene together with a functionally redundant close homolog that we name BOP2. The BOP genes are members of a gene family containing ankyrin repeats and a BTB/POZ domain, suggesting a role in protein-protein interaction. We show that the BOP genes are expressed in the proximal parts of plant lateral organs where they repress the transcription not only of class 1 knox genes but also of JAG. We also show that the BOP genes are acting together with the flower meristem identity gene LEAFY in the suppression of bract formation. These findings show that the BOP genes are important regulators of the growth and development of lateral organs. PMID- 15800003 TI - The homeobox transcription factor Barx2 regulates chondrogenesis during limb development. AB - Among the many factors involved in regulation of chondrogenesis, bone morphogenetic proteins (BMPs) and members of the Sox and homeobox transcription factor families have been shown to have crucial roles. Of these regulators, the homeobox transcription factors that function during chondrogenesis have been the least well defined. We show here that the homeobox transcription factor Barx2 is expressed in primary mesenchymal condensations, digital rays, developing joints and articular cartilage of the developing limb, suggesting that it plays a role in chondrogenesis. Using retroviruses and antisense oligonucleotides to manipulate Barx2 expression in limb bud micromass cultures, we determined that Barx2 is necessary for mesenchymal aggregation and chondrogenic differentiation. In accordance with these findings, Barx2 regulates the expression of several genes encoding cell-adhesion molecules and extracellular matrix proteins, including NCAM and collagen II (Col2a1) in the limb bud. Barx2 bound to elements within the cartilage-specific Col2a1 enhancer, and this binding was reduced by addition of Barx2 or Sox9 antibodies, or by mutation of a HMG box adjacent to the Barx2-binding element, suggesting cooperation between Barx2 and Sox proteins. Moreover, both Barx2 and Sox9 occupy Col2a1 enhancer during chondrogenesis in vivo. We also found that two members of the BMP family that are crucial for chondrogenesis, GDF5 and BMP4, regulate the pattern of Barx2 expression in developing limbs. Based on these data, we suggest that Barx2 acts downstream of BMP signaling and in concert with Sox proteins to regulate chondrogenesis. PMID- 15800004 TI - Valois, a component of the nuage and pole plasm, is involved in assembly of these structures, and binds to Tudor and the methyltransferase Capsuleen. AB - Using the Capsuleen (Csul) methyltransferase as bait in the yeast two-hybrid system, we have identified a novel Drosophila protein containing multiple WD repeats and encoded by the valois (vsl) gene, which acts in pole plasm function. Vls is homologous to human MEP50, which forms a complex with the PRMT5 methyltransferase--the human homologue of Csul. We found that Vls localizes to the nuage in the nurse cells and to the pole plasm in the oocyte. Moreover vls is required for the synthesis and/or stability of Oskar and the localization of Tudor (Tud) in both the nuage and at the posterior pole of the oocyte. Furthermore, we show that Vls and a fragment of Tud interact directly in binding assay. As the PMRT5/MEP50 complex is involved in ribonucleoprotein complex assembly, we hypothesize that the Vls complex may play a similar function in assembling the nuage in nurse cells and the polar granules in the oocyte. PMID- 15800005 TI - A comparison of the diagnostic accuracy and prognostic value of the first and second anti-cyclic citrullinated peptides (CCP1 and CCP2) autoantibody tests for rheumatoid arthritis. AB - OBJECTIVES: To compare the diagnostic performance and prognostic value of the anti-cyclic citrullinated peptide (CCP1) and anti-CCP2 autoantibody tests in a clinical setting. METHODS: Anti-CCP1 and anti-CCP2 antibody tests were performed on the same serum samples obtained from 467 patients with early arthritis from the Leiden Arthritis Cohort. The sensitivity, specificity, positive predictive value, and negative predictive value for discriminating between rheumatoid arthritis (RA) and non-RA at 1 year's follow up were calculated for both tests. Results were graphically presented using receiver operating characteristic curves. Progression of radiological joint damage was assessed over 4 years in patients with RA and used to assess the prognostics values of the CCP tests. RESULTS: At a similar specificity the CCP2 test had a higher sensitivity than the CCP1 test. Both tests identified a subgroup of patients with RA with an increased rate of joint damage progression. The anti-CCP2 test identified more patients with an increased rate of joint damage progression than the anti-CCP1 test, and in multiple regression analysis CCP2 was the better predictor of joint damage. CONCLUSIONS: The CCP2 test had better diagnostic and prognostic ability than the CCP1 test. PMID- 15800006 TI - Differential effect of IL10 and TNF{alpha} genotypes on determining susceptibility to discoid and systemic lupus erythematosus. AB - OBJECTIVE: To ascertain the possible involvement of functional interleukin 10 (IL10) and tumour necrosis alpha (TNFalpha) cytokine promoter polymorphisms on the susceptibility to discoid and systemic lupus erythematosus (DLE, SLE), and their associations with immunological features. METHODS: Single nucleotide polymorphisms of the IL10 (-1082, -819, and -592) and TNFalpha (-308) genes were determined using allele specific probes in 248 lupus patients and 343 matched controls. To assess functional significance of genotypes, basal mRNA cytokine levels were quantified in 106 genotyped healthy controls by real time RT-PCR. Specific autoantibodies and cutaneous manifestations were analysed in SLE patients and associated with functional genotypes. RESULTS: After analysing the distribution of IL10 and TNFalpha transcript levels according to promoter genotypes in healthy individuals, patients and controls were classified into functional single and combined genotypes according to the expected high or low constitutive cytokine production. High TNFalpha genotypes (-308AA or AG) were associated with SLE independently of IL10 alleles, whereas the risk of developing DLE and the prevalence of discoid lesion in SLE were higher in the high IL10/low TNFalpha producer group (-1082GG/-308GG). Cytokine interaction also influences the appearance of autoantibodies. Antibodies against Sm are prevalent among low producer patients for both cytokines, a genotype not associated with lupus incidence, whereas low IL10/high TNFalpha patients have the highest frequency of antibodies to SSa and SSb. CONCLUSIONS: IL10/TNFalpha interaction influences susceptibility to DLE and the appearance of specific autoantibodies in SLE patients, whereas high TNFalpha producer genotypes represent a significant risk factor for SLE. PMID- 15800007 TI - Direct costs related to rheumatoid arthritis: the patient perspective. AB - OBJECTIVE: To determine rheumatoid arthritis related out of pocket expenditure (OOPE) in Germany and to disaggregate the total OOPE into contributing cost domains. METHODS: Data for the cost analysis were drawn from a multicentre randomised controlled prospective trial to assess the effectiveness of clinical quality management in patients with rheumatoid arthritis. Both payer sources and patient cost questionnaires were used to generate health care utilisation data. All cost domains of a recently published matrix were reviewed and potential sources of OOPE were determined. Health care utilisation data were developed throughout 2001. Co-payment regulations as per January 2004 were applied in order to indicate the most recent level of OOPE in Germany. Data were analysed in both physical and monetary units using descriptive statistics. RESULTS: In all, 136 patients with rheumatoid arthritis were included. Mean total OOPE per patient and year was 417.20 Euro (SEM 38.8, median 271.2). OOPE accounted for 15.3% of the total direct costs of rheumatoid arthritis. Total OOPE were further subdivided into cost domains: "non-physician service utilisation"' (194.40 Euro per patient and year; SEM 24.2), "medication" (99.00 Euro; 6.1), "transportation" (56.20 Euro; 17.4), "visits to physicians" (38.40 Euro; 0.6), "hospital facilities" (24.00 Euro; 5.6), and "devices and aids" (5.10 Euro; 0.8). CONCLUSIONS: Rheumatoid arthritis is associated with substantial OOPE, imposing a considerable economic burden for patients. OOPE contribute significantly to the total health care expenditure in rheumatoid arthritis. The patient perspective has to be taken into account when calculating the overall direct costs of rheumatoid arthritis from a societal point of view. PMID- 15800009 TI - A comparison of two primary care trials on tennis elbow: issues of external validity. AB - OBJECTIVE: To assess clinical heterogeneity across two studies with respect to study population, interventions, and outcome measures, and to evaluate the influence of these sources of heterogeneity on the results of the studies. METHODS: The individual patient data were used from two randomised controlled trials investigating the effectiveness of conservative treatments in patients with tennis elbow in primary care. Patients were allocated at random to treatment with steroid injection, wait and see policy, non-steroidal anti-inflammatory drugs, placebo tablets, or physiotherapy. Outcome measures included severity of the main complaint, inconvenience of the elbow complaints, pain during the day, elbow disability, pain-free grip strength, and global improvement. All outcomes were assessed at 1, 6, and 12 months after randomisation. RESULTS: The two study populations were similar with respect to age, sex, comorbid neck/shoulder complaints, and baseline scores for the severity of pain. However, significant differences were observed for employment status, duration of elbow complaints, dominant side affected, previous history of elbow complaints, and use of analgesics. Local injections differed between the two studies with respect to volume, number, and steroid preparation. However, after 1, 6, and 12 months, the treatment effects of steroid injections were very similar between the study populations. CONCLUSIONS: Despite large differences in study population at baseline, the responses to steroid injections were remarkably similar. Also the responses to other conservative interventions and the placebo treatment were very consistent, suggesting a uniform course of a tennis elbow and a lack of influence of clinical heterogeneity. PMID- 15800008 TI - Hypoxia inducible factor (HIF) in rheumatology: low O2! See what HIF can do! AB - Maintenance of oxygen homoeostasis is the basic principle in cell proliferation, differentiation, survival, and function in all higher organisms. The transcription factor, HIF (hypoxia inducible factor) has a central role in oxygen homoeostasis, and is indispensably linked to energy metabolism. Abnormally reduced oxygen concentrations leading to dysfunctional cell metabolism are found in rheumatoid arthritis and hence, knowledge of the molecular adaptive responses to hypoxia and the involvement of HIF in the pathogenesis of RA are interesting. PMID- 15800010 TI - Prevalence of rheumatoid arthritis in France: 2001. AB - BACKGROUND: Prevalence estimates of rheumatoid arthritis (RA) vary across Europe. Recent estimates in southern European countries showed a lower prevalence than in northern countries. OBJECTIVES: To estimate the prevalence of RA in France in a multiregional representative sample in the year 2001. METHODS: A two stage random sample was constituted in seven areas (20 counties) from the national telephone directory of households and by the next birthday method in each household. Patient-interviewers, member of self help groups, were trained to administer telephone surveys using a validated questionnaire for case detection of inflammatory rheumatism, and conducted the survey under quality control. All suspected cases of RA were confirmed by their rheumatologist or by clinical examination. Prevalence estimates after probability sampling correction were standardised for age and sex (national census 1999). RESULTS: An average response rate of 64.7% (two stages combined) led to a total of 9395 respondents. Standardised prevalence was 0.31% (95% confidence interval 0.18 to 0.48) for RA, 0.51% in women and 0.09% in men, with a higher age-specific prevalence in the 65 74 year age band. A geographical analysis of county clustering showed significant variation across the country. CONCLUSION: This national multiregional cooperative study demonstrates the usefulness of working in association with patients of self help groups. It showed a similar prevalence of RA to that of the spondyloarthropathies estimated concomitantly during the survey. It provides a reliable basis for definition of population targets for healthcare delivery and drug treatments. PMID- 15800011 TI - Limited proteolysis differentially modulates the stability and subcellular localization of domains of RPGRIP1 that are distinctly affected by mutations in Leber's congenital amaurosis. AB - The retinitis pigmentosa GTPase regulator (RPGR) protein interacts with the retinitis pigmentosa GTPase regulator interacting protein-1 (RPGRIP1). Genetic lesions in the cognate genes lead to distinct and severe human retinal dystrophies. The biological role of these proteins in retinal function and pathogenesis of retinal diseases is elusive. Here, we present the first physiological assay of the role of RPGRIP1 and mutations therein. We found that the monoallelic and homozygous mutations, DeltaE1279 and D1114G, in the RPGR interacting domain (RID) of RPGRIP1, enhance and abolish, respectively, its interaction in vivo with RPGR without affecting the stability of RID. In contrast to RID(WT) and RID(D1114G), chemical genetics shows that the interaction of RID(DeltaE1279) with RPGR is resistant to various stress treatments such as osmotic, pH and heat-shock stimuli. Hence, RID(D1114G) and RID(DeltaE1279) constitute loss- and gain-of-function mutations. Moreover, we find that the isoforms, bRPGRIP1 and bRPGRIP1b, undergo limited proteolysis constitutively in vivo in the cytoplasm compartment. This leads to the relocation and accumulation of a small and stable N-terminal domain of approximately 7 kDa to the nucleus, whereas the cytosolic C-terminal domain of RPGRIP1 is degraded and short-lived. The RID(D1114G) and RID(DeltaE1279) mutations exhibit strong cis-acting and antagonistic biological effects on the nuclear relocation, subcellular distribution and proteolytic cleavage of RPGRIP1 and/or domains thereof. These data support distinct and spatiotemporal subcellular-specific roles to RPGRIP1. A novel RPGRIP1-mediated nucleocytoplasmic crosstalk and transport pathway regulated by RID, and hence by RPGR, emerges with implications in the molecular pathogenesis of retinopathies, and a model to other diseases. PMID- 15800012 TI - SNPs, microarrays and pooled DNA: identification of four loci associated with mild mental impairment in a sample of 6000 children. AB - Mild mental impairment (MMI) represents the low extreme of the quantitative trait of general intelligence and is highly heritable. Quantitative trait loci (QTLs) conferring susceptibility to MMI, as for most complex traits, are likely to be of small effect size. Using a novel approach we call SNP-MaP (SNP Microarrays and Pooling), we have identified four loci associated with MMI. These four loci have been replicated in two SNP-MaP studies and verified by individual genotyping. The two SNP-MaP studies conducted were a case versus control comparison (n = 515 and n = 1028, respectively) and a low versus high general intelligence extremes group comparison (n = 503 and n = 505, respectively). Each of the four groups consisted of five independent 'subpools', with each subpool assayed on a separate microarray. Twelve loci showing the largest significant differences in both SNP MaP studies were individually genotyped on 6154 children. Of the four loci positively associated with MMI, the minor allele of each conferred the greater risk for MMI. Two of the loci are close to known genes and may be in linkage disequilibrium with them. One of the loci is between the candidate genes KLF7 and CREB1, but given possible long-range effects on expression and the unknown importance of untranslated elements such as micro-RNAs, all four loci deserve attention as candidates. Although each SNP accounts for a small amount of variance, their effects are additive and they can be combined in a 'SNP set' that can be used as a genetic risk index for MMI in behavioral genomic analyses. PMID- 15800013 TI - Decreased expression of ABCD4 and BG1 genes early in the pathogenesis of X-linked adrenoleukodystrophy. AB - Childhood cerebral adrenoleukodystrophy (CCER), adrenomyeloneuropathy (AMN) and AMN with cerebral demyelination (AMN-C) are the main phenotypic variants of X linked adrenoleukodystrophy (ALD). It is caused by mutations in the ABCD1 gene encoding a half-size peroxisomal transporter that has to dimerize to become functional. The biochemical hallmark of ALD is the accumulation of very-long chain fatty acids (VLCFA) in plasma and tissues. However, there is no correlation between the ALD phenotype and the ABCD1 gene mutations or the accumulation of VLCFA in plasma and fibroblast from ALD patients. The absence of genotype phenotype correlation suggests the existence of modifier genes. To elucidate the mechanisms underlying the phenotypic variability of ALD, we studied the expression of ABCD1, three other peroxisomal transporter genes of the same family (ABCD2, ABCD3 and ABCD4) and two VLCFA synthetase genes (VLCS and BG1) involved in VLCFA metabolism, as well as the VLCFA concentrations in the normal white matter (WM) from ALD patients with CCER, AMN-C and AMN phenotypes. This study shows that: (1) ABCD1 gene mutations leading to truncated ALD protein are unlikely to cause variation in the ALD phenotype; (2) accumulation of saturated VLCFA in normal-appearing WM correlates with ALD phenotype and (3) expression of the ABCD4 and BG1, but not of the ABCD2, ABCD3 and VLCS genes, tends to be correlated with the severity of the disease, acting early in the pathogenesis of ALD. PMID- 15800014 TI - Functional analysis of Peutz-Jeghers mutations reveals that the LKB1 C-terminal region exerts a crucial role in regulating both the AMPK pathway and the cell polarity. AB - Germline mutations of the LKB1 gene are responsible for the cancer-prone Peutz Jeghers syndrome (PJS). LKB1 encodes a serine-threonine kinase that acts as a regulator of cell cycle, metabolism and cell polarity. The majority of PJS missense mutations abolish LKB1 enzymatic activity and thereby impair all functions assigned to LKB1. Here, we have investigated the functional consequences of recurrent missense mutations identified in PJS and in sporadic tumors which map in the LKB1 C-terminal non-catalytic region. We report that these C-terminal mutations neither disrupt LKB1 kinase activity nor interfere with LKB1-induced growth arrest. However, these naturally occuring mutations lessened LKB1-mediated activation of the AMP-activated protein kinase (AMPK) and impaired downstream signaling. Furthermore, C-terminal mutations compromise LKB1 ability to establish and maintain polarity of both intestinal epithelial cells and migrating astrocytes. Consistent with these findings, mutational analysis reveals that the LKB1 tail exerts an essential function in the control of cell polarity. Overall, our results ascribe a crucial regulatory role to the LKB1 C terminal region. Our findings further indicate that LKB1 tumor suppressor activity is likely to depend on the regulation of AMPK signaling and cell polarization. PMID- 15800015 TI - Pathogenic effects of a novel heterozygous R350P desmin mutation on the assembly of desmin intermediate filaments in vivo and in vitro. AB - Mutations of the human desmin gene on chromosome 2q35 cause a familial or sporadic form of skeletal myopathy frequently associated with cardiac abnormalities. Here, we report the pathogenic effects of a novel heterozygous R350P desmin missense mutation, which resides in the evolutionary highly conserved coil 2B domain of the alpha-helical coiled-coil desmin rod domain, on the assembly of desmin intermediate filaments (IF) in cultured cells and in vitro. By transfection experiments, we show that R350P desmin is incapable of de novo formation of a desmin IF network in vimentin-free BMGE+H, MCF7 and SW13 cells and that it disrupts the endogenous vimentin cytoskeleton in 3T3 fibroblast cells. Hence, transfected cells displayed abnormal cytoplasmic protein aggregates reminiscent of desmin-positive protein deposits seen in the immunohistochemical and ultrastructural analysis of skeletal muscle derived from the index patient of the affected family. To study the functional effects of the R350P desmin mutation at the protein level, we performed in vitro assembly studies with wild-type (WT) and mutant desmin protein. Our analysis revealed that the in vitro assembly process of R350P desmin is already disturbed at the unit length filament level and that further association reactions generate huge, tightly packed protein aggregates. On assessing the pathogenic effects of R350P desmin in various mixtures with WT desmin, we show that a ratio of 1 : 3 (R350P desmin/WT desmin) is sufficient to effectively block the normal polymerization process of desmin IFs. Our findings indicate that the heterozygous R350P desmin mutation exerts a dominant negative effect on the ordered lateral arrangement of desmin subunits. This disturbance of the lateral packing taking place in the first phase of assembly is ultimately leading to abnormal protein aggregation. PMID- 15800016 TI - Brain natriuretic peptide in heart failure: an improving prognosis? PMID- 15800017 TI - Increased circulating C-reactive protein and macrophage-colony stimulating factor are complementary predictors of long-term outcome in patients with chronic coronary artery disease. AB - AIMS: We investigated, in a 6 year follow-up study, whether circulating levels of C-reactive protein (CRP) and macrophage colony stimulating factor (MCSF) have an independent or complementary prognostic value in patients with chronic coronary artery disease (CAD). METHODS AND RESULTS: MCSF and CRP were measured in 100 patients with chronic CAD. Of 95 (33%) patients, 31 who completed the 6 year follow-up presented adverse events (death, myocardial infarction, and unstable angina). In multivariable analysis (including traditional risk factors and medications), the upper tertiles of MCSF (> or =814 pg/mL) and CRP (> or =2.5 mg/L) levels were independently associated with a 13- and 6-fold increase in risk of events, respectively (P<0.01). Patients with combined high CRP and MCSF had a higher absolute risk of events than patients with elevated MCSF or CRP alone (75 vs. 59 vs. 32%, respectively, P<0.01). The mean event-free time was 39, 64, and 52 months in patients with elevated MCSF, elevated CRP, and their combination, respectively. CONCLUSION: In patients with chronic CAD, the prognostic value of MCSF is independent and complementary to that of CRP. MCSF is a particularly useful prognostic marker when CRP levels are low, but also provides additional information concerning risk and time-course of events in patients with elevated CRP. PMID- 15800018 TI - Are even impaired fasting blood glucose levels preoperatively associated with increased mortality after CABG surgery? AB - AIMS: Impaired fasting glucose (IFG) below the diagnostic threshold for diabetes mellitus (DM) is associated with macrovascular pathology and increased mortality after percutaneous coronary interventions. The study goal was to determine whether pre-operative fasting blood glucose (fB-glu) is associated with an increased mortality after coronary artery bypass grafting (CABG). METHODS AND RESULTS: During 2001-03, 1895 patients underwent primary CABG [clinical DM (CDM) in 440/1895; complete data on fB-glu for n=1375/1455]. Using pre-operative fB glu, non-diabetics were categorized as having normal fB-glu (<5.6 mmol/L), IFG (5.6< or =fB-glu<6.1 mmol/L), or suspected DM (SDM) (> or =6.1 mmol/L). fB-glu was normal in 59%. The relative risks of 30 day and 1 year mortality compared with patients with normal fB-glu was 1.7 [95% confidence interval (CI): 0.5-5.5] and 2.9 (CI: 0.8-11.2) with IFG, 2.8 (CI: 1.1-7.2) and 1.9 (CI: 0.5-6.3) with SDM vs. 1.8 (CI: 0.8-4.0) and 1.6 (CI: 0.6-4.3) if CDM, respectively. The receiver operator characteristic area for the continuous variable fB-glu and 1 year mortality was 0.65 (P=0.002). CONCLUSION: The elevated risk of death after CABG surgery known previously to be associated with CDM seems also to be shared by a group of similar size that includes patients with IFG and undiagnosed DM. PMID- 15800019 TI - Prognostic implication of activated partial thromboplastin time after reteplase or half-dose reteplase plus abciximab: results from the GUSTO-V trial. AB - AIMS: To evaluate the relationship between activated partial thromboplastin time (aPTT) and clinical outcomes in the Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO-V) trial comparing standard-dose reteplase to half-dose reteplase and abciximab. METHODS AND RESULTS: We analysed data on 11,420 patients receiving unfractionated heparin. Peak aPTT levels recorded during the hospitalization were correlated with clinical outcomes. Multivariable logistic regression models examined the relationship between peak aPTT levels and (i) moderate-to-severe bleeding, (ii) intracerebral haemorrhage, (iii) reinfarction, and (iv) 30-day mortality. Non-linear relationships were explored in the models using cubic spline functions. Higher rates of significant complications were seen in both groups when aPTT levels were <50 s or when levels were >70 s. In the combination therapy group, the relationship between aPTT levels and bleeding appeared accentuated. Reinfarction rates increased gradually as aPTT levels were >70 s in both groups, but the relationships were not statistically significant. Peak aPTT levels <50 s were associated with increased 30-day mortality even after multivariable adjustment. CONCLUSION: Peak aPTT levels <50 s and >70 s are associated with worse clinical outcomes in the modern era of fibrinolytic therapy; these relationships are different in patients receiving standard reteplase vs. combination therapy. PMID- 15800020 TI - Advanced heart failure: feasibility study of long-term continuous axial flow pump support. AB - AIMS: A lack of donor hearts has stimulated interest in using blood pumps to treat severe heart failure. We tested the hypothesis that a new continuous flow circulatory assist device could be employed safely to relieve symptoms of heart failure and evaluated the potential to prolong life. METHODS AND RESULTS: An intracardiac axial flow pump was implanted in 17 heart failure patients [idiopathic dilated (12), ischaemic (4), or amyloid cardiomyopathy (1)]. All were deemed ineligible for transplantation. Implantation of the device was by left thoracotomy (15) or median sternotomy (2). Power delivery was by a skull-mounted titanium pedestal. All patients survived surgery. None needed right ventricular support. There were three hospital deaths, two early from subdural haematoma and aortic thrombosis, one late after switching to transplantation. A total of 14 patients left hospital with a cumulative support-time of 15.9 years (median: 293 days, interquartile range: 286 days, 1-44 months). Actuarial 1-, 2-, and 3-year survivals were 56, 47, and 24%, respectively. There was no pump failure. Quality of life scores improved. Two superficial pedestal infections were successfully treated. Four patients had cerebral thrombo-embolism: two early events attributed to inadequate anticoagulation and two late with near-complete resolution. An improved anticoagulant regime addressed this problem. Late death occurred in five patients from battery disconnection, subdural haematoma, bowel ischaemia, respiratory failure, and after cardiac transplantation. CONCLUSION: Continuous flow blood pumps provided symptomatic relief of severe heart failure with high quality of life. Event-free survival reached 4 years. Analysis of adverse events led to improved management strategies. There is potential for widespread use of blood pumps in the community. A controlled trial is required. PMID- 15800021 TI - Neuroprotection by inhibition of matrix metalloproteinases in a mouse model of intracerebral haemorrhage. AB - Intracerebral haemorrhage (ICH) is an acute neurological disorder without effective treatment. Mechanisms of acute brain injury after ICH remain to be clarified. Although a few studies suggested a detrimental role for the gelatinase matrix metalloproteinase (MMP)-9 in ICH, the relationship between MMP-9 activity and acute brain injury after ICH is not determined. In this study, we first examined the expression of gelatinases in vivo using a collagenase-induced mouse model of ICH. Gel zymography revealed that MMP-9 was activated and upregulated after ICH. In situ zymography showed that gelatinase activity was mostly co localized with neurons and endothelial cells of the blood vessel matrix. Inhibition with a broad-spectrum metalloproteinase inhibitor GM6001 (100 mg/kg) ameliorated dysregulated gelatinase activity, neutrophil infiltration, production of oxidative stress, brain oedema and degenerating neurons. Functional improvement and a decrease in injury volume were also observed. We provide evidence that MMP-9 may play a deleterious role in acute brain injury within the first 3 days after ICH. Blockade of MMP activity during this critical period may have efficacy as a therapeutic strategy for the treatment of acute brain injury after ICH. PMID- 15800022 TI - Short-lived plasma blasts are the main B cell effector subset during the course of multiple sclerosis. AB - Multiple sclerosis is a chronic inflammatory and demyelinating disorder of the CNS with an unknown aetiology. Although intrathecal immunoglobulin G (IgG) synthesis is a key feature of the disease, little is still known about the B cell response in the CNS of multiple sclerosis patients. We analysed the phenotype and kinetics of different B cell subsets in patients with multiple sclerosis, infectious disease (IND) and non-inflammatory neurological disease (NIND). B cells were detected in the CSF of multiple sclerosis and IND patients, but were largely absent in NIND patients. In the CSF, the majority of B cells had a phenotype of memory B cells and short-lived plasma blasts (PB); plasma cells were absent from the compartment. The proportion of PB was highest in multiple sclerosis patients and patients with acute CNS infection. While PB disappeared rapidly from the CSF after resolution of infection in IND patients, these cells were present at high numbers throughout the disease course in multiple sclerosis patients. CSF PB numbers in multiple sclerosis patients strongly correlated with intrathecal IgG synthesis and inflammatory parenchymal disease activity as disclosed by MRI. This study identifies short-lived plasma blasts as the main effector B cell population involved in ongoing active inflammation in multiple sclerosis patients. PMID- 15800023 TI - Visuo-spatial consciousness and parieto-occipital areas: a high-resolution EEG study. AB - Conscious and unconscious visuo-spatial processes are mainly related to parieto occipital cortical activation. In this study, the working hypothesis was that a specific pattern of parieto-occipital activation is induced by conscious, as opposed to unconscious, visuo-spatial processes. Electroencephalographic data (128 channels) were recorded in 12 normal adults during a visuo-spatial task. A cue stimulus appeared on the right or the left (equal probability) monitor side for a 'threshold time' inducing approximately 50% of correct recognitions. It was followed (after 2 s) by visual go stimuli at spatially congruent or incongruent positions with reference to the cue location. The left (right) mouse button was clicked if the go stimulus appeared on the left (right) monitor side. Subjects were required to say 'seen' if they had detected the cue stimulus or 'not seen' if they missed it (self-report). 'Seen' and 'not seen' electroencephalographic trials were averaged separately to form visual evoked potentials. Sources of these potentials were estimated by LORETA software. Reaction time to go stimuli was shorter during spatially congruent than incongruent 'seen' trials, possibly due to covert attention on cue for self-report. It was also shorter during spatially congruent than incongruent 'not seen' trials, as an objective sign of unconscious processes. Cue stimulus evoked parieto-occipital potentials which has the same peak latencies in the 'seen' and 'not seen' cases. Sources of these potentials were located in occipital area 19 and parietal area 7. Source strength was significantly stronger in 'seen' than 'not seen' cases at approximately +300 ms post-stimulus. These results may unveil features of parieto-occipital activation accompanying visuo-spatial consciousness. PMID- 15800024 TI - Serial and parallel processing in the human auditory cortex: a magnetoencephalographic study. AB - Although anatomical, histochemical and electrophysiological findings in both animals and humans have suggested a parallel and serial mode of auditory processing, precise activation timings of each cortical area are not well known, especially in humans. We investigated the timing of arrival of signals to multiple cortical areas using magnetoencephalography in humans. Following click stimuli applied to the left ear, activations were found in six cortical areas in the right hemisphere: the posteromedial part of Heschl's gyrus (HG) corresponding to the primary auditory cortex (PAC), the anterolateral part of the HG region on or posterior to the transverse sulcus, the posterior parietal cortex (PPC), posterior and anterior parts of the superior temporal gyrus (STG), and the planum temporale (PT). The mean onset latencies of each cortical activity were 17.1, 21.2, 25.3, 26.2, 30.9 and 47.6 ms respectively. These results suggested a serial model of auditory processing along the medio-lateral axis of the supratemporal plane and, in addition, implied the existence of several parallel streams running postero-superiorly (from the PAC to the belt region and then to the posterior STG, PPC or PT) and anteriorly (PAC-belt-anterior STG). PMID- 15800025 TI - A mutually stimulating loop involving emx2 and canonical wnt signalling specifically promotes expansion of occipital cortex and hippocampus. AB - The correct size of the different areas composing the mature cerebral cortex depends on the proper early allocation of cortical progenitors to their distinctive areal fates, as well as on appropriate subsequent tuning of their area-specific proliferation-differentiation profiles. Whereas much is known about the genetics of the former process, the molecular mechanisms regulating proliferation and differentiation rates within distinctive cortical proto-areas are still largely obscure. Here we show that a mutual stimulating loop, involving Emx2 and canonical Wnt signalling, specifically promotes expansion of the occipito-hippocampal anlage. Collapse of this loop occurring in Emx2-/- mutants leads progenitors within this region to slow down DNA synthesis and exit prematurely from the cell cycle, due to misregulation of cell cycle-, proneural- and lateral inhibition-molecular machineries, and eventually results in dramatic and selective size-reduction of occipital cortex and hippocampus. Reactivation of canonical Wnt signalling in the same mutants rescues a subset of molecular abnormalities and corrects differentiation rates of occipito-hippocampal progenitors. PMID- 15800026 TI - Principles governing auditory cortex connections. AB - Topographic maps are common constituents of the primary auditory, visual, and somatic sensory cortex. However, in most cortical areas, no such maps have yet been identified, posing a conceptual problem for theories of cortical function centered on topography. What principle guides the organization of these other areas? We investigated this issue in cat auditory cortex. The connectional topography of five tonotopic areas and eight non-tonotopic areas was assessed using retrograde tract tracing and quantified by three metrics: clustering, dispersion, and separation. Clustering measures the spatial density of labeled neurons, dispersion provides an index of their spread, and separation serves as a scaling metric. These parameters each show that all auditory cortical regions receive precise and equally topographic connections from thalamic, corticocortical, and commissural sources. This isotropic principle suggests a common substrate for coordinating communication across the cortex and may reflect common mechanisms related to the developmental patterning of connections. This unifying principle extends to auditory and prefrontal cortex, and perhaps to other neocortical areas. PMID- 15800027 TI - Synergistic effect of SCF and TNF-alpha on the up-regulation of cell-surface expression of ICAM-1 on human leukemic mast cell line (HMC)-1 cells. AB - Intercellular adhesion molecule-1 (ICAM-1) has been shown to play crucial roles in mast cell interaction with other inflammatory cells and recruitment into the inflamed tissue. In the present study, human mast cell line-1 (HMC-1) was stimulated with different cytokines including stem cell factor (SCF), tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-13, IL-18, and IL-25. Cell surface expression of ICAM-1 was assessed by flow cytometry. To elucidate the intracellular signal transduction regulating the ICAM-1 expression, phosphorylated extracellular signal-regulated kinase (ERK), phosphorylated p38 mitogen-activated protein kinase (MAPK), and nuclear factor (NF)-kappaB translocation were assessed by enzyme-linked immunosorbent assay. Results showed that SCF, TNF-alpha, and IL-13 but not IL-18 and IL-25 could up-regulate the surface expression of ICAM-1 on HMC-1 cells. A synergistic effect of SCF and TNF alpha on ICAM-1 expression was demonstrated. This synergistic effect was shown to be dose-dependently enhanced by SCF but not TNF-alpha. Results indicated that SCF activated ERK, and TNF-alpha activated the p38 MAPK and NF-kappaB pathway. Selective inhibitor of ERK, PD098059, and c-kit inhibitors, STI571 and PP1, suppressed the combined SCF and TNF-alpha-induced ICAM-1 expression. BAY117082 but not SB203580, which are the inhibitors of NF-kappaB and p38 MAPK, respectively, suppressed the TNF-alpha-induced ICAM-1 expression. Therefore, SCF and TNF-alpha acted through ERK and the NF-kappaB pathway to regulate the ICAM-1 expression and elicited the synergistic effect. In conclusion, our results provide insight for cross-talk between different signaling pathways that can help in understanding the fine control of adhesion molecule expression under the concerted effects of cytokines. PMID- 15800028 TI - Expression of the common heat-shock protein receptor CD91 is increased on monocytes of exposed yet HIV-1-seronegative subjects. AB - The significantly higher surface expression of the surface heat-shock protein receptor CD91 on monocytes of human immunodeficiency virus type-1 (HIV-1) infected, long-term nonprogressors suggests that HIV-1 antigen uptake and cross presentation mediated by CD91 may contribute to host anti-HIV-1 defenses and play a role in protection against HIV-1 infection. To investigate this further, we performed phenotypic analysis to compare CD91 surface expression on CD14(+) monocytes derived from a cohort of HIV-1-exposed seronegative (ESN) subjects, their seropositive (SP) partners, and healthy HIV-1-unexposed seronegative (USN) subjects. The median fluorescent intensity (MFI) of CD91 on CD14(+) monocytes was significantly higher in ESN compared with SP (P = 0.028) or USN (P = 0.007), as well as in SP compared with USN subjects (P = 0.018). CD91 MFI was not normalized in SP subjects on highly active antiretroviral therapy (HAART) despite sustainable, undetectable plasma viraemia. Data in three SP subjects experiencing viral rebounds following interruption of HAART showed low CD91 MFI comparable with levels in USN subjects. There was a significant positive correlation between CD91 MFI and CD8(+) T cell counts in HAART-naive SP subjects (r = 0.7, P = 0.015). Increased surface expression of CD91 on CD14(+) monocytes is associated with the apparent HIV-1 resistance that is observed in ESN subjects. PMID- 15800029 TI - Production of matrix metalloproteinase-9 by activated human monocytes involves a phosphatidylinositol-3 kinase/Akt/IKKalpha/NF-kappaB pathway. AB - Matrix metalloproteinase-9 (MMP-9) is considered to be an important component in the progression of inflammation. Monocytes/macrophages are prominent at inflammation sites, and activation of these cells by stimulants, such as lipopolysaccharide (LPS) or tumor necrosis factor alpha and granulocyte macrophage-colony stimulating factor, leads to the production of significant amounts of MMP-9. Here, we show that LPS stimulation of monocytes results in MMP 9 production through a phosphatidylinositol-3 kinase (PI-3K)/Akt/inhibitor of kappaB (IkappaB) kinase-alpha (IKKalpha)/nuclear factor (NF)-kappaB pathway. This new role for Akt in signaling leading to MMP-9 production was demonstrated by inhibitor and immunoprecipitation studies. LY294002 or wortmannin, inhibitors of PI-3K, suppressed LPS-induced Akt activity and MMP-9 production. Evidence for the participation of Akt in monocyte MMP-9 synthesis was demonstrated by the inhibition of MMP-9 by SH-5, a specific inhibitor of Akt. The mechanism by which Akt regulates MMP-9 is through the activation of NF-kappaB, as shown by coimmunoprecipitation of the phosphorylated form of IKKalpha and Akt as well as the SH-5 suppression of the dissociation of IkappaB from NF-kappaB and the activation of NF-kappaB p65. The role of NF-kappaB in regulation of MMP-9 was demonstrated further by the inhibition of MMP-9 production by proteasome inhibitors, lactacystin and MG-132, which prevented the ubiquitination and dissociation of IkappaB from NF-kappaB. This is the first demonstration that Akt is involved in the signaling pathway leading to the production of monocyte MMP-9 and provides an additional approach in the regulation of this enzyme in human primary monocytes. PMID- 15800030 TI - 2,3,5-tris(Glutathion-S-yl)hydroquinone (TGHQ)-mediated apoptosis of human promyelocytic leukemia cells is preceded by mitochondrial cytochrome c release in the absence of a decrease in the mitochondrial membrane potential. AB - 2,3,5-tris(Glutathion-S-yl)hydroquinone (TGHQ), a metabolite of benzene, induces apoptosis in human promyelocytic leukemia (HL-60) cells. However, the mechanisms by which TGHQ induces apoptosis are unclear, and they were the focus of the present investigation. TGHQ stimulated the rapid formation (30 min) of reactive oxygen species (ROS) in HL-60 cells, and co-treatment with catalase or the antioxidant N-acetylcysteine (NAC) completely blocked TGHQ-induced apoptosis, implicating a causative role for ROS in HL-60 cell death. Western blot analysis revealed the complete disappearance of pro-caspase 9 between 1 and 2 hours after exposure of HL-60 cells to TGHQ, concomitant with the appearance of cleaved caspase 9 and increases in caspase 9 activity. The appearance of two cleaved forms of caspase 3 occurred subsequent to increases in caspase 9 activity. Levels of the anti-apoptotic Bcl-2 protein remained constant during TGHQ-induced apoptosis of HL-60 cells, but Bcl-2 S70 phosphorylation decreased. In contrast, changes in the subcellular localization of the pro-apoptotic molecule Bax were observed, with a rapid (15-60 min) increase in the ratio of cytosolic to mitochondrial Bax. Cytochrome c release from mitochondria to the cytosol occurred after Bax translocation and the dephosphorylation of pS70 Bcl-2. However the mitochondrial inner transmembrane potential (deltapsi(m)) was maintained, even after cytochrome c was released from the mitochondria. Cyclosporin A, an inhibitor of the mitochondrial membrane permeability transition pore (PTP), did not completely rescue HL-60 cells from apoptosis. Taken together, we conclude that TGHQ facilitates ROS production, alters the post-translational modification of Bcl-2 and subcellular localization of Bax, culminating in the release of cytochrome c and caspase activation. PMID- 15800031 TI - Enhancement of cyanide-induced mitochondrial dysfunction and cortical cell necrosis by uncoupling protein-2. AB - Uncoupling protein 2 (UCP-2) is expressed in the inner mitochondrial membrane and modulates mitochondrial function by partially uncoupling oxidative phosphorylation, and it has been reported to modulate cell death. Cyanide is a potent neurotoxin that inhibits complex IV to alter mitochondrial function to induce neuronal death. In primary rat cortical cells KCN produced an apoptotic death at 200-400 microM. Higher concentrations of potassium cyanide (KCN) (500 600 microM) switched the mode of death from apoptosis to necrosis. In necrotic cells, ATP levels were severely depleted as compared to cortical cells undergoing apoptosis. To determine if UCP-2 expression could alter KCN-induced cell death, cells were transiently transfected with full-length human UCP-2 cDNA (UCP-2+). Overexpression switched the mode of death produced by KCN (400 microM) from apoptosis to necrosis. The change in cell death was mediated by impaired mitochondrial function as reflected by a marked decrease of ATP levels and reduction in mitochondrial membrane potential. RNA interference or transfection with a dominant interfering mutant blocked the necrotic response observed in UCP 2+ cells. Additionally, treatment of UCP-2+ cells with cyclosporin A blocked necrosis, indicating the involvement of mitochondrial permeability pore transition in the necrotic death. These results show that increased expression of UCP-2 alters the response to a potent mitochondrial toxin by switching the mode of cell death from apoptosis to necrosis. It is concluded that UCP-2 levels influence cellular responses to cyanide-induced mitochondrial dysfunction. PMID- 15800032 TI - Neurotoxicological and statistical analyses of a mixture of five organophosphorus pesticides using a ray design. AB - Environmental exposures generally involve chemical mixtures instead of single chemicals. Statistical models such as the fixed-ratio ray design, wherein the mixing ratio (proportions) of the chemicals is fixed across increasing mixture doses, allows for the detection and characterization of interactions among the chemicals. In this study, we tested for interaction(s) in a mixture of five organophosphorus (OP) pesticides (chlorpyrifos, diazinon, dimethoate, acephate, and malathion). The ratio of the five pesticides (full ray) reflected the relative dietary exposure estimates of the general population as projected by the US EPA Dietary Exposure Evaluation Model (DEEM). A second mixture was tested using the same dose levels of all pesticides, but excluding malathion (reduced ray). The experimental approach first required characterization of dose-response curves for the individual OPs to build a dose-additivity model. A series of behavioral measures were evaluated in adult male Long-Evans rats at the time of peak effect following a single oral dose, and then tissues were collected for measurement of cholinesterase (ChE) activity. Neurochemical (blood and brain cholinesterase [ChE] activity) and behavioral (motor activity, gait score, tail pinch response score) endpoints were evaluated statistically for evidence of additivity. The additivity model constructed from the single chemical data was used to predict the effects of the pesticide mixture along the full ray (10-450 mg/kg) and the reduced ray (1.75-78.8 mg/kg). The experimental mixture data were also modeled and statistically compared to the additivity models. Analysis of the 5-OP mixture (the full ray) revealed significant deviation from additivity for all endpoints except tail-pinch response. Greater-than-additive responses (synergism) were observed at the lower doses of the 5-OP mixture, which contained non-effective dose levels of each of the components. The predicted effective doses (ED20, ED50) were about half that predicted by additivity, and for brain ChE and motor activity, there was a threshold shift in the dose-response curves. For the brain ChE and motor activity, there was no difference between the full (5 OP mixture) and reduced (4-OP mixture) rays, indicating that malathion did not influence the non-additivity. While the reduced ray for blood ChE showed greater deviation from additivity without malathion in the mixture, the non-additivity observed for the gait score was reversed when malathion was removed. Thus, greater-than-additive interactions were detected for both the full and reduced ray mixtures, and the role of malathion in the interactions varied depending on the endpoint. In all cases, the deviations from additivity occurred at the lower end of the dose-response curves. PMID- 15800033 TI - Temporal and dose-dependent hepatic gene expression patterns in mice provide new insights into TCDD-Mediated hepatotoxicity. AB - In an effort to further characterize the mechanisms of 2,3,7,8-tetrachlorodibenzo p-dioxin (TCDD)-mediated toxicity, comprehensive temporal and dose-response microarray analyses were performed on hepatic tissue from immature ovariectomized C57BL/6 mice treated with TCDD. For temporal analysis, mice were gavaged with 30 microg/kg of TCDD or vehicle and sacrificed after 2, 4, 8, 12, 18, 24, 72, or 168 h. Dose-response mice were gavaged with 0, 0.001, 0.01, 0.1, 1, 10, 100, or 300 microg/kg of TCDD and sacrificed after 24 h. Hepatic gene expression profiles were monitored using custom cDNA microarrays containing 13,362 cDNA clones. Gene expression analysis identified 443 and 315 features which exhibited a significant change at one or more doses or time points, respectively, as determined using an empirical Bayes approach. Functional gene annotation extracted from public databases associated gene expression changes with physiological processes such as oxidative stress and metabolism, differentiation, apoptosis, gluconeogenesis, and fatty acid uptake and metabolism. Complementary histopathology (H&E and Oil Red O stains), clinical chemistry (i.e., alanine aminotransferase [ALT], triglyceride [TG], free fatty acids [FFA], cholesterol) and high-resolution gas chromatography/mass spectrometry assessment of hepatic TCDD levels were also performed in order to phenotypically anchor changes in gene expression to physiological end points. Collectively, the data support a proposed mechanism for TCDD-mediated hepatotoxicity, including fatty liver, which involves mobilization of peripheral fat and inappropriate increases in hepatic uptake of fatty acids. PMID- 15800034 TI - Supplement to the Carcinogenic Potency Database (CPDB): results of animal bioassays published in the general literature through 1997 and by the National Toxicology Program in 1997-1998. AB - The Carcinogenic Potency Database (CPDB) is a systematic and unifying resource that standardizes the results of chronic, long-term animal cancer tests which have been conducted since the 1950s. The analyses include sufficient information on each experiment to permit research into many areas of carcinogenesis. Both qualitative and quantitative information is reported on positive and negative experiments that meet a set of inclusion criteria. A measure of carcinogenic potency, TD(50) (daily dose rate in mg/kg body weight/day to induce tumors in half of test animals that would have remained tumor-free at zero dose), is estimated for each tissue-tumor combination reported. This article is the ninth publication of a chronological plot of the CPDB; it presents results on 560 experiments of 188 chemicals in mice, rats, and hamsters from 185 publications in the general literature updated through 1997, and from 15 Reports of the National Toxicology Program in 1997-1998. The test agents cover a wide variety of uses and chemical classes. The CPDB Web Site (http://potency.berkeley.edu/) presents the combined database of all published plots in a variety of formats as well as summary tables by chemical and by target organ, supplemental materials on dosing and survival, a detailed guide to using the plot formats, and documentation of methods and publications. The overall CPDB, including the results in this article, presents easily accessible results of 6153 experiments on 1485 chemicals from 1426 papers and 429 NCI/NTP (National Cancer Institute/National Toxicology program) Technical Reports. A tab-separated format of the full CPDB for reading the data into spreadsheets or database applications is available on the Web Site. PMID- 15800035 TI - The refinement of uncertainty/safety factors in risk assessment by the incorporation of data on toxicokinetic variability in humans. AB - The derivation of safe levels of exposure in humans for compounds that are assumed to cause threshold toxicity has relied on the application of a 100-fold uncertainty factor to a measure for the threshold, such as the no observed adverse effect level (NOAEL) or the benchmark dose (BMD). This 100-fold safety factor consists of the product of two 10-fold factors allowing for human variability and interspecies differences. The International Programme on Chemical Safety has suggested the subdivision of these 10-fold factors to allow for variability in toxicokinetics and toxicodynamics. This subdivision allows the replacement of the default uncertainty factors with a chemical-specific adjustment factor (CSAF) when suitable data are available. This short review describes potential options to refine safety factors used in risk assessment, with particular emphasis on pathway-related uncertainty factors associated with variability in kinetics. These pathway-related factors were derived from a database that quantified interspecies differences and human variability in phase I metabolism, phase II metabolism, and renal excretion. This approach allows metabolism and pharmacokinetic data in healthy adults and subgroups of the population to be incorporated in the risk-assessment process and constitutes an intermediate approach between simple default factors and chemical-specific adjustment factors. PMID- 15800036 TI - Functional genomic analysis of the rates of protein evolution. AB - The evolutionary rates of proteins vary over several orders of magnitude. Recent work suggests that analysis of large data sets of evolutionary rates in conjunction with the results from high-throughput functional genomic experiments can identify the factors that cause proteins to evolve at such dramatically different rates. To this end, we estimated the evolutionary rates of >3,000 proteins in four species of the yeast genus Saccharomyces and investigated their relationship with levels of expression and protein dispensability. Each protein's dispensability was estimated by the growth rate of mutants deficient for the protein. Our analyses of these improved evolutionary and functional genomic data sets yield three main results. First, dispensability and expression have independent, significant effects on the rate of protein evolution. Second, measurements of expression levels in the laboratory can be used to filter data sets of dispensability estimates, removing variates that are unlikely to reflect real biological effects. Third, structural equation models show that although we may reasonably infer that dispensability and expression have significant effects on protein evolutionary rate, we cannot yet accurately estimate the relative strengths of these effects. PMID- 15800037 TI - Color-blind fluorescence detection for four-color DNA sequencing. AB - We present an approach called pulsed multiline excitation (PME) for measurements of multicomponent, fluorescence species and demonstrate its application in capillary electrophoresis for DNA sequencing. To fully demonstrate the advantages of PME, a fluorescent dye set has been developed whose absorption maxima span virtually the entire visible spectrum. Unlike emission wavelength-dependent approaches for identifying fluorescent species, the removal of the spectral component in PME confers a number of advantages including higher and normalized signals from all dyes present in the assay, the elimination of spectral cross talk between dyes, and higher signal collection efficiency. Base-calling is unambiguously determined once dye mobility corrections are made. These advantages translate into significantly enhanced signal quality as illustrated in the primary DNA sequencing data and provide a means for achieving accurate base calling at lower reagent concentrations. PMID- 15800038 TI - Decline in skeletal muscle mitochondrial function with aging in humans. AB - Cumulative mtDNA damage occurs in aging animals, and mtDNA mutations are reported to accelerate aging in mice. We determined whether aging results in increased DNA oxidative damage and reduced mtDNA abundance and mitochondrial function in skeletal muscle of human subjects. Studies performed in 146 healthy men and women aged 18-89 yr demonstrated that mtDNA and mRNA abundance and mitochondrial ATP production all declined with advancing age. Abundance of mtDNA was positively related to mitochondrial ATP production rate, which in turn, was closely associated with aerobic capacity and glucose tolerance. The content of several mitochondrial proteins was reduced in older muscles, whereas the level of the oxidative DNA lesion, 8-oxo-deoxyguanosine, was increased, supporting the oxidative damage theory of aging. These results demonstrate that age-related muscle mitochondrial dysfunction is related to reduced mtDNA and muscle functional changes that are common in the elderly. PMID- 15800039 TI - Conformational transition of amyloid beta-peptide. AB - The amyloid beta-peptides (Abetas), containing 39-43 residues, are the key protein components of amyloid deposits in Alzheimer's disease. To structurally characterize the dynamic behavior of Abeta(40), 12 independent long-time molecular dynamics (MD) simulations for a total of 850 ns were performed on both the wide-type peptide and its mutant in both aqueous solution and a biomembrane environment. In aqueous solution, an alpha-helix to beta-sheet conformational transition for Abeta(40) was observed, and an entire unfolding process from helix to coil was traced by MD simulation. Structures with beta-sheet components were observed as intermediates in the unfolding pathway of Abeta(40). Four glycines (G(25), G(29), G(33), and G(37)) are important for Abeta(40) to form beta-sheet in aqueous solution; mutations of these glycines to alanines almost abolished the beta-sheet formation and increased the content of the helix component. In the dipalmitoyl phosphatidylcholine (DPPC) bilayer, the major secondary structure of Abeta(40) is a helix; however, the peptide tends to exit the membrane environment and lie down on the surface of the bilayer. The dynamic feature revealed by our MD simulations rationalized several experimental observations for Abeta(40) aggregation and amyloid fibril formation. The results of MD simulations are beneficial to understanding the mechanism of amyloid formation and designing the compounds for inhibiting the aggregation of Abeta and amyloid fibril formation. PMID- 15800040 TI - Modeling gene and genome duplications in eukaryotes. AB - Recent analysis of complete eukaryotic genome sequences has revealed that gene duplication has been rampant. Moreover, next to a continuous mode of gene duplication, in many eukaryotic organisms the complete genome has been duplicated in their evolutionary past. Such large-scale gene duplication events have been associated with important evolutionary transitions or major leaps in development and adaptive radiations of species. Here, we present an evolutionary model that simulates the duplication dynamics of genes, considering genome-wide duplication events and a continuous mode of gene duplication. Modeling the evolution of the different functional categories of genes assesses the importance of different duplication events for gene families involved in specific functions or processes. By applying our model to the Arabidopsis genome, for which there is compelling evidence for three whole-genome duplications, we show that gene loss is strikingly different for large-scale and small-scale duplication events and highly biased toward certain functional classes. We provide evidence that some categories of genes were almost exclusively expanded through large-scale gene duplication events. In particular, we show that the three whole-genome duplications in Arabidopsis have been directly responsible for >90% of the increase in transcription factors, signal transducers, and developmental genes in the last 350 million years. Our evolutionary model is widely applicable and can be used to evaluate different assumptions regarding small- or large-scale gene duplication events in eukaryotic genomes. PMID- 15800041 TI - Quantitative prediction of perceptual decisions during near-threshold fear detection. AB - A fundamental goal of cognitive neuroscience is to explain how mental decisions originate from basic neural mechanisms. The goal of the present study was to investigate the neural correlates of perceptual decisions in the context of emotional perception. To probe this question, we investigated how fluctuations in functional MRI (fMRI) signals were correlated with behavioral choice during a near-threshold fear detection task. fMRI signals predicted behavioral choice independently of stimulus properties and task accuracy in a network of brain regions linked to emotional processing: posterior cingulate cortex, medial prefrontal cortex, right inferior frontal gyrus, and left insula. We quantified the link between fMRI signals and behavioral choice in a whole-brain analysis by determining choice probabilities by means of signal-detection theory methods. Our results demonstrate that voxel-wise fMRI signals can reliably predict behavioral choice in a quantitative fashion (choice probabilities ranged from 0.63 to 0.78) at levels comparable to neuronal data. We suggest that the conscious decision that a fearful face has been seen is represented across a network of interconnected brain regions that prepare the organism to appropriately handle emotionally challenging stimuli and that regulate the associated emotional response. PMID- 15800042 TI - Evidence for the presence of Mn(III) intermediates in the bacterial oxidation of Mn(II). AB - Bacterial oxidation of Mn(II) to Mn(IV) is believed to drive the oxidative segment of the global biogeochemical Mn cycle and regulates the concentration of dissolved Mn(II) in the oceanic water column, where it is a critical nutrient for planktonic primary productivity. Mn(II) oxidizing activity is expressed by numerous phylogenetically diverse bacteria and fungi, suggesting that it plays a fundamental and ubiquitous role in the environment. This important redox system is believed to be driven by an enzyme or enzyme complex involving a multicopper oxidase, although the biochemical mechanism has never been conclusively demonstrated. Here, we show that Mn(II) oxidation by spores of the marine Bacillus sp. strain SG-1 is a result of two sequential one-step electron transfer processes, both requiring the putative multicopper oxidase, MnxG, in which Mn(III) is a transient intermediate. A kinetic model of the oxidation pathway is presented, which shows that the Mn(II) to Mn(III) step is the rate-limiting step. Thus, oxidation of Mn(II) appears to involve a unique multicopper oxidase system capable of the overall two-electron oxidation of its substrate. This enzyme system may serve as a source for environmental Mn(III), a strong oxidant and competitor for siderophore-bound Fe(III) in nutrient-limited environments. That metabolically dormant spores catalyze an important biogeochemical process intimately linked to the C, N, Fe, and S cycles requires us to rethink the role of spores in the environment. PMID- 15800043 TI - Transcriptome analysis of the aphid bacteriocyte, the symbiotic host cell that harbors an endocellular mutualistic bacterium, Buchnera. AB - Aphids possess bacteriocytes, cells specifically differentiated to harbor obligatory mutualistic bacteria of the genus Buchnera, which have lost many genes that are essential for common bacterial functions. To understand the host's role in maintaining the symbiotic relationship, bacteriocytes were isolated from the pea aphid, Acyrthosiphon pisum, and the host transcriptome was investigated by using EST analysis and real-time quantitative RT-PCR. A number of genes were highly expressed specifically in the bacteriocyte, including (i) genes for amino acid metabolism, including those for biosynthesis of amino acids that Buchnera cannot produce, and those for utilization of amino acids that Buchnera can synthesize; (ii) genes related to transport, including genes for mitochondrial transporters and a gene encoding Rab, a G protein that regulates vesicular transport; and (iii) genes for putative lysozymes that degrade bacterial cell walls. Significant up-regulation of i clearly indicated that the bacteriocyte is involved in the exchange of amino acids between the host aphid and Buchnera, the key metabolic process in the symbiotic system. Conspicuously high expression of ii and iii shed light on previously unknown aspects of the host-Buchnera interactions in the symbiotic system. PMID- 15800044 TI - Protein folding pathways from replica exchange simulations and a kinetic network model. AB - We present an approach to the study of protein folding that uses the combined power of replica exchange simulations and a network model for the kinetics. We carry out replica exchange simulations to generate a large ( approximately 10(6)) set of states with an all-atom effective potential function and construct a kinetic model for folding, using an ansatz that allows kinetic transitions between states based on structural similarity. We use this network to perform random walks in the state space and examine the overall network structure. Results are presented for the C-terminal peptide from the B1 domain of protein G. The kinetics is two-state after small temperature perturbations. However, the coil-to-hairpin folding is dominated by pathways that visit metastable helical conformations. We propose possible mechanisms for the alpha-helix/beta-hairpin interconversion. PMID- 15800045 TI - Simulations of the pressure and temperature unfolding of an alpha-helical peptide. AB - We study by molecular simulations the reversible folding/unfolding equilibrium as a function of density and temperature of a solvated alpha-helical peptide. We use an extension of the replica exchange molecular dynamics method that allows for density and temperature Monte Carlo exchange moves. We studied 360 thermodynamic states, covering a density range from 0.96 to 1.14 g.cm(-3) and a temperature range from 300 to 547.6 K. We simulated 10 ns per replica for a total simulation time of 3.6 micros. We characterize the structural, thermodynamic, and hydration changes as a function of temperature and pressure. We also calculate the compressibility and expansivity of unfolding. We find that pressure does not affect the helix-coil equilibrium significantly and that the volume change upon pressure unfolding is small and negative (-2.3 ml/mol). However, we find significant changes in the coordination of water molecules to the backbone carbonyls. This finding predicts that changes in the chemical shifts and IR spectra with pressure can be due to changes in coordination and not only changes in the helical content. A simulation of the IR spectrum shows that water coordination effects on frequency shifts are larger than changes due to elastic structural changes in the peptide. PMID- 15800046 TI - Glutamate receptor activation triggers a calcium-dependent and SNARE protein dependent release of the gliotransmitter D-serine. AB - The gliotransmitter D-serine is released upon (S)-alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionic acid/kainate and metabotropic glutamate receptor stimulation, but the mechanisms involved are unknown. Here, by using a highly sensitive bioassay to continuously monitor extracellular D-serine levels, we have investigated the pathways used in its release. We reveal that D-serine release is inhibited by removal of extracellular calcium and augmented by increasing extracellular calcium or after treatment with the Ca(2+) ionophore A23187. Furthermore, release of the amino acid is considerably reduced after depletion of thapsigargin-sensitive intracellular Ca(2+) stores or chelation of intracellular Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetate-acetoxymethyl ester. Interestingly, D-serine release also was markedly reduced by concanamycin A, a vacuolar-type H(+)-ATPase inhibitor, indicating a role for the vesicular proton gradient in the transmitter storage/release. In addition, agonist-evoked D serine release was sensitive to tetanus neurotoxin. Finally, immunocytochemical and sucrose density gradient analysis revealed that a large fraction of D-serine colocalized with synaptobrevin/VAMP2, suggesting that it is stored in VAMP2 bearing vesicles. In summary, our study reveals the cellular mechanisms subserving D-serine release and highlights the importance of the glial cell exocytotic pathway in influencing CNS levels of extracellular D-serine. PMID- 15800047 TI - Kaposi's sarcoma-associated herpesvirus expresses an array of viral microRNAs in latently infected cells. AB - MicroRNAs (miRNAs) are an endogenously encoded class of small RNAs that have been proposed to function as key posttranscriptional regulators of gene expression in a range of eukaryotic species, including humans. The small size of miRNA precursors makes them potentially ideal for use by viruses as inhibitors of host cell defense pathways. Here, we demonstrate that the pathogenic human herpesvirus Kaposi's sarcoma-associated herpesvirus (KSHV) encodes an array of 11 distinct miRNAs, all of which are expressed at readily detectable levels in latently KSHV infected cells. Individual KSHV miRNAs were expressed at up to 2,200 copies per cell. The KSHV miRNAs are expressed from what appears to be a single genetic locus that largely coincides with an approximately 4-kb noncoding sequence located between the KSHV v-cyclin and K12/Kaposin genes, both of which are also expressed in latently infected cells. Computer analysis of potential mRNA targets for these viral miRNAs identified a number of interesting candidate genes, including several mRNAs previously shown to be down-regulated in KSHV-infected cells. We hypothesize that these viral miRNAs play a critical role in the establishment and/or maintenance of KSHV latent infection in vivo and, hence, in KSHV-induced oncogenesis. PMID- 15800049 TI - Developmental expression and biophysical characterization of a Drosophila melanogaster aquaporin. AB - Aquaporins (AQPs) accelerate the movement of water and other solutes across biological membranes, yet the molecular mechanisms of each AQP's transport function and the diverse physiological roles played by AQP family members are still being defined. We therefore have characterized an AQP in a model organism, Drosophila melanogaster, which is amenable to genetic manipulation and developmental analysis. To study the mechanism of Drosophila Malpighian tubule (MT)-facilitated water transport, we identified seven putative AQPs in the Drosophila genome and found that one of these, previously named DRIP, has the greatest sequence similarity to those vertebrate AQPs that exhibit the highest rates of water transport. In situ mRNA analyses showed that DRIP is expressed in both embryonic and adult MTs, as well as in other tissues in which fluid transport is essential. In addition, the pattern of DRIP expression was dynamic. To define DRIP-mediated water transport, the protein was expressed in Xenopus oocytes and in yeast secretory vesicles, and we found that significantly elevated rates of water transport correlated with DRIP expression. Moreover, the activation energy required for water transport in DRIP-expressing secretory vesicles was 4.9 kcal/mol. This low value is characteristic of AQP-mediated water transport, whereas the value in control vesicles was 16.4 kcal/mol. In contrast, glycerol, urea, ammonia, and proton transport were unaffected by DRIP expression, suggesting that DRIP is a highly selective water-specific channel. This result is consistent with the homology between DRIP and mammalian water-specific AQPs. Together, these data establish Drosophila as a new model system with which to investigate AQP function. PMID- 15800048 TI - A contact-activated kinase signals Candida albicans invasive growth and biofilm development. AB - For mammalian cells, contact-dependent regulatory controls are crucially important for controlling cellular proliferation and preventing diseases such as cancer. Candida albicans, an opportunistic fungal pathogen that normally resides within a mammalian host, also exhibits contact-dependent cellular behaviors such as invasive hyphal growth and biofilm development. Results reported here demonstrate that, in C. albicans, physical contact results in activation of the mitogen-activated protein kinase Mkc1p. This kinase is part of the fungal cell integrity pathway, a signal transduction pathway known to be activated by cell wall stress. It is demonstrated here that Mkc1p is required for invasive hyphal growth and normal biofilm development. Therefore, Mkc1p signaling contributes to contact-dependent regulation. Because responding to contact appropriately allows coordinated cellular behavior in a metazoan, commensal C. albicans cells behave like a part of the host, using contact-activated signaling to regulate fungal behavior. PMID- 15800050 TI - RNA interference targeted to multiple P2X receptor subtypes attenuates zinc induced calcium entry. AB - A postulated therapeutic avenue in cystic fibrosis (CF) is activation of Ca(2+) dependent Cl(-) channels via stimulation of Ca(2+) entry from extracellular solutions independent of CFTR functional status. We have shown that extracellular zinc and ATP induce a sustained increase in cytosolic Ca(2+) in human airway epithelial cells that translates into stimulation of sustained secretory Cl(-) transport in non-CF and CF human and mouse airway epithelial cells, cell monolayers, and nasal mucosa. On the basis of these studies, the Ca(2+) entry channels most likely involved were P2X purinergic receptor channels. In the present study, molecular and biochemical data show coexpression of P2X(4), P2X(5), and P2X(6) subtypes in non-CF (16HBE14o(-)) and CF (IB3-1) human bronchial epithelial cells. Other P2X receptor Ca(2+) entry channel subtypes are expressed rarely or not at all in airway epithelia, epithelial cell models from other CF-relevant tissues, or vascular endothelia. Novel transient lipid transfection-mediated delivery of small interference RNA fragments specific to P2X(4) and P2X(6) (but not P2X(5)) into IB3-1 CF human airway epithelial cells inhibited extracellular zinc- and ATP-induced Ca(2+) entry markedly in fura-2 Ca(2+) measurements and "knocked down" protein by >65%. These data suggest that multiple P2X receptor Ca(2+) entry channel subtypes are expressed in airway epithelia. P2X(4) and P2X(6) may coassemble on the airway surface as targets for possible therapeutics for CF independent of CFTR genotype. PMID- 15800051 TI - Direct block of cloned hKv1.5 channel by cytochalasins, actin-disrupting agents. AB - The action of cytochalasins, actin-disrupting agents on human Kv1.5 channel (hKv1.5) stably expressed in Ltk(-) cells was investigated using the whole cell patch-clamp technique. Cytochalasin B inhibited hKv1.5 currents rapidly and reversibly at +60 mV in a concentration-dependent manner with an IC(50) of 4.2 microM. Cytochalasin A, which has a structure very similar to cytochalasin B, inhibited hKv1.5 (IC(50) of 1.4 microM at +60 mV). Pretreatment with other actin filament disruptors cytochalasin D and cytochalasin J, and an actin filament stabilizing agent phalloidin had no effect on the cytochalasin B-induced inhibition of hKv1.5 currents. Cytochalasin B accelerated the decay rate of inactivation for the hKv1.5 currents. Cytochalasin B-induced inhibition of the hKv1.5 channels was voltage dependent with a steep increase over the voltage range of the channel's opening. However, the inhibition exhibited voltage independence over the voltage range in which channels are fully activated. Cytochalasin B produced no significant effect on the steady-state activation or inactivation curves. The rate constants for association and dissociation of cytochalasin B were 3.7 microM/s and 7.5 s(-1), respectively. Cytochalasin B produced a use-dependent inhibition of hKv1.5 current that was consistent with the slow recovery from inactivation in the presence of the drug. Cytochalasin B (10 microM) also inhibited an ultrarapid delayed rectifier K(+) current (I(K,ur)) in human atrial myocytes. These results indicate that cytochalasin B primarily blocks activated hKv1.5 channels and endogenous I(K,ur) in a cytoskeleton independent manner as an open-channel blocker. PMID- 15800052 TI - Changes in intracellular Ca2+ and pH in response to thapsigargin in human glioblastoma cells and normal astrocytes. AB - Despite extensive work in the field of glioblastoma research no significant increase in survival rates for this devastating disease has been achieved. It is known that disturbance of intracellular Ca(2+) ([Ca(2+)](i)) and intracellular pH (pH(i)) regulation could be involved in tumor formation. The sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) is a major regulator of [Ca(2+)](i). We have investigated the effect of inhibition of SERCA by thapsigargin (TG) on [Ca(2+)](i) and pH(i) in human primary glioblastoma multiforme (GBM) cells and GBM cell lines, compared with normal human astrocytes, using the fluorescent indicators fura-2 and BCECF, respectively. Basal [Ca(2+)](i) was higher in SK-MG 1 and U87 MG but not in human primary GBM cells compared with normal astrocytes. However, in tumor cells, TG evoked a much larger and faster [Ca(2+)](i) increase than in normal astrocytes. This increase was prevented in nominally Ca(2+)-free buffer and by 2-APB, an inhibitor of store-operated Ca(2+) channels. In addition, TG-activated Ca(2+) influx, which was sensitive to 2-APB, was higher in all tumor cell lines and primary GBM cells compared with normal astrocytes. The pH(i) was also elevated in tumor cells compared with normal astrocytes. TG caused acidification of both normal and all GBM cells, but in the tumor cells, this acidification was followed by an amiloride- and 5-(N,N-hexamethylene)-amiloride sensitive recovery, indicating involvement of a Na(+)/H(+) exchanger. In summary, inhibition of SERCA function revealed a significant divergence in intracellular Ca(2+) homeostasis and pH regulation in tumor cells compared with normal human astrocytes. PMID- 15800053 TI - HAI-1 regulates activation and expression of matriptase, a membrane-bound serine protease. AB - Hepatocyte growth factor activator inhibitor-1 (HAI-1) was initially identified as cognate inhibitor of matriptase, a membrane-bound serine protease. Paradoxically, HAI-1 is also required for matriptase activation, a process that requires sphingosine 1-phosphate (S1P)-mediated translocation of the protease to cell-cell junctions in human mammary epithelial cells. In the present study, we further explored how HAI-1 regulates this protease. First, we observed that after S1P treatment HAI-1 was cotranslocated with matriptase to cell-cell junctions and that the cellular ratio of HAI-1 to matriptase was maintained during this process. However, when this ratio was changed by cell treatment with HAI-1 small interfering RNA or anti-HAI-1 MAb M19, spontaneous activation of matriptase occurred in the absence of S1P-induced translocation; S1P-induced matriptase activation was also enhanced. These results support a role for HAI-1 in protection of cell from uncontrolled matriptase activation. We next expressed matriptase, either alone or with HAI-1 in breast cancer cells that do not endogenously express either protein. A defect in matriptase trafficking to the cell surface occurred if wild-type matriptase was expressed in the absence of HAI 1; this defect appeared to result from matriptase toxicity to cells. Coexpression with matriptase of wild-type HAI-1, but not HAI-1 mutants altered in its Kunitz domain 1, corrected the trafficking defect. In contrast, catalytically defective matriptase mutants were normal in their trafficking in the absence of HAI-1. These results are also consistent with a role for HAI-1 to prevent inappropriate matriptase proteolytic activity during its protein synthesis and trafficking. Taken together, these results support multiple roles for HAI-1 to regulate matriptase, including its proper expression, intracellular trafficking, activation, and inhibition. PMID- 15800054 TI - Gender dimorphism in rat liver mitochondrial oxidative metabolism and biogenesis. AB - In the present study, we have investigated gender differences in rat liver mitochondrial oxidative metabolism. Total mitochondrial population (M) as well as the heavy (M1), medium (M3), and light (M8) mitochondrial fractions obtained by means of differential centrifugation steps at 1,000, 3,000, and 8,000 g, respectively, were isolated. Electron microscopic analysis was performed and mitochondrial protein content and cardiolipin levels, mitochondrial O(2) flux, ATP synthase activity, mitochondrial membrane potential, and mitochondrial transcription factor A (TFAM) protein levels were measured in each sample. Our results indicate that mitochondria from females have higher protein content and higher cardiolipin levels, greater respiratory and phosphorylative capacities, and more-energized mitochondria in respiratory state 3. Moreover, protein levels of TFAM were four times greater in females than in males. Gender differences in the aforementioned parameters were more patent in the isolated heavy M1 and M3 mitochondrial fractions. The present study demonstrates that gender-related differences in liver mitochondrial function are due mainly to a higher capacity and efficiency of substrate oxidation, likely related to greater mitochondrial machinery in females than in males, which is in accord with greater mitochondrial differentiation in females. PMID- 15800055 TI - SLC26A9 is expressed in gastric surface epithelial cells, mediates Cl-/HCO3- exchange, and is inhibited by NH4+. AB - HCO3- secretion by gastric mucous cells is essential for protection against acidic injury and peptic ulcer. Herein we report the identification of an apical HCO3- transporter in gastric surface epithelial cells. Northern hybridization and RT-PCR demonstrate the expression of this transporter, also known as SLC26A9, in mouse and rat stomach and trachea (but not kidney). In situ hybridization in mouse stomach showed abundant expression of SLC26A9 in surface epithelial cells with apical localization on immunofluorescence labeling. Functional studies in HEK-293 cells demonstrated that SLC26A9 mediates Cl-/HCO3- exchange and is also capable of Cl--independent HCO3- extrusion. Unlike other anion exchangers or transport proteins reported to date, SLC26A9 activity is inhibited by ammonium (NH4+). The inhibitory effect of NH4+ on gastric HCO3- secretion was also indicated by reduced gastric juxtamucosal pH (pHjm) in rat stomach in vivo. This report is the first to describe the inhibition of HCO3- transport in vitro and the reduction of pHjm in stomach in vivo by NH4+. Given its critical localization on the apical membrane of surface epithelial cells, its ability to transport HCO3 , and its inhibition by NH4+, we propose that SLC26A9 mediates HCO3- secretion in surface epithelial cells and is essential for protection against acidic injury in the stomach. Disease states that are associated with increased ammonia (NH3)/NH4+ generation (e.g., Helicobacter pylori) may impair gastric HCO3- secretion and therefore predispose patients to peptic ulcer by inhibiting SLC26A9. PMID- 15800056 TI - Insulin-activated, K+-channel-sensitive Akt pathway is primary mediator of ML-1 cell proliferation. AB - Voltage-gated K(+) channel activities are involved in regulating growth factor stimulated cell proliferation in a variety of cell types. Here we report that suppression of a voltage-gated K(+) channel with 4-aminopyridine (4-AP), barium, and tetraethylammonium inhibited both EGF- and insulin-stimulated myeloblastic leukemia ML-1 cell proliferation in a concentration-dependent manner. Both MAPK/ERK and Akt pathways are known to mediate cell proliferative signals of a variety of growth factors including insulin. In serum-starved ML-1 cells, insulin rapidly stimulated phosphorylation of ERK1/2 and Akt, and the phosphorylation levels peaked approximately 30 min after treatment. Pretreatment of ML-1 cells with 4-AP potently and dose-dependently prevented phosphorylation of ERK1/2 and Akt. However, insulin-induced activation of the Akt pathway also played a role in promoting ML-1 cell proliferation. Flow cytometry analysis revealed that although ML-1 cells were primarily arrested at G(1) phase by serum starvation for 36 h, they reentered the cell cycle after treatment with serum or insulin for 24 h. However, concomitant 4-AP treatment was able to attenuate cell cycle progression in synchronized ML-1 cells stimulated with growth factors. Our results strongly suggest that a 4-AP-sensitive K(+) channel activity plays an important role in controlling proliferation of ML-1 cells by affecting the activation of multiple signal transduction processes induced by insulin. PMID- 15800057 TI - Arginine vasopressin stimulation of cerebral microvascular endothelial cell Na-K Cl cotransporter activity is V1 receptor and [Ca] dependent. AB - Ischemia-induced brain edema formation is mediated by increased transport of Na and Cl across an intact blood-brain barrier (BBB). Our previous studies have provided evidence that a luminally located BBB Na-K-Cl cotransporter is stimulated during cerebral ischemia to increase transport of Na and Cl into the brain. The main focus of the present study was to evaluate the effects of arginine vasopressin (AVP), previously shown to be increased in the brain during ischemia and to promote edema formation, on activity of the BBB cotransporter. Cerebral microvascular endothelial cell (CMEC) monolayers were cultured in astroglial cell conditioned medium, and Na-K-Cl cotransporter activity was assessed as bumetanide-sensitive (86)Rb influx. In both human and bovine CMECs, as well as in freshly isolated microvessels, AVP stimulated cotransport activity. This stimulatory effect was mimicked by V(1) but not V(2) vasopressin agonists and was blocked by V(1) but not V(2) vasopressin antagonists. Consistent with a V(1) vasopressin receptor mechanism of action, AVP caused an increase in CMEC intracellular [Ca] that was blocked by a V(1) antagonist. Exposing the cells to [Ca]-free media and/or reducing intracellular [Ca] by BAPTA also blocked AVP stimulation of CMEC cotransporter activity, as did the phospholipase C inhibitor U-73122. Finally, we found that while stimulation of CMEC cotransporter activity by AVP occurred within minutes, it was also sustained for hours in the continued presence of AVP. These findings support the hypothesis that AVP, through a V(1) receptor- and [Ca]-dependent mechanism, stimulates the BBB Na-K-Cl cotransporter to participate in ischemia-induced edema formation. PMID- 15800058 TI - p115 Interacts with the GLUT4 vesicle protein, IRAP, and plays a critical role in insulin-stimulated GLUT4 translocation. AB - Insulin-regulated aminopeptidase (IRAP) is an abundant cargo protein of Glut4 storage vesicles (GSVs) that traffics to and from the plasma membrane in response to insulin. We used the amino terminus cytoplasmic domain of IRAP, residues 1 109, as an affinity reagent to identify cytosolic proteins that might be involved in GSV trafficking. In this way, we identified p115, a peripheral membrane protein known to be involved in membrane trafficking. In murine adipocytes, we determined that p115 was localized to the perinuclear region by immunofluorescence and throughout the cell by fractionation. By immunofluorescence, p115 partially colocalizes with GLUT4 and IRAP in the perinuclear region of cultured fat cells. The amino terminus of p115 binds to IRAP and overexpression of a N-terminal construct results in its colocalization with GLUT4 throughout the cell. Insulin-stimulated GLUT4 translocation is completely inhibited under these conditions. Overexpression of p115 C-terminus has no significant effect on GLUT4 distribution and translocation. Finally, expression of the p115 N-terminus construct has no effect on the distribution and trafficking of GLUT1. These data suggest that p115 has an important and specific role in insulin-stimulated Glut4 translocation, probably by way of tethering insulin-sensitive Glut4 vesicles at an as yet unknown intracellular site. PMID- 15800059 TI - Dictyostelium LIS1 is a centrosomal protein required for microtubule/cell cortex interactions, nucleus/centrosome linkage, and actin dynamics. AB - The widespread LIS1-proteins were originally identified as the target for sporadic mutations causing lissencephaly in humans. Dictyostelium LIS1 (DdLIS1) is a microtubule-associated protein exhibiting 53% identity to human LIS1. It colocalizes with dynein at isolated, microtubule-free centrosomes, suggesting that both are integral centrosomal components. Replacement of the DdLIS1 gene by the hypomorphic D327H allele or overexpression of an MBP-DdLIS1 fusion disrupted various dynein-associated functions. Microtubules lost contact with the cell cortex and were dragged behind an unusually motile centrosome. Previously, this phenotype was observed in cells overexpressing fragments of dynein or the XMAP215 homologue DdCP224. DdLIS1 was coprecipitated with DdCP224, suggesting that both act together in dynein-mediated cortical attachment of microtubules. Furthermore, DdLIS1-D327H mutants showed Golgi dispersal and reduced centrosome/nucleus association. Defects in DdLIS1 function also altered actin dynamics characterized by traveling waves of actin polymerization correlated with a reduced F-actin content. DdLIS1 could be involved in actin dynamics through Rho-GTPases, because DdLIS1 interacted directly with Rac1A in vitro. Our results show that DdLIS1 is required for maintenance of the microtubule cytoskeleton, Golgi apparatus and nucleus/centrosome association, and they suggest that LIS1-dependent alterations of actin dynamics could also contribute to defects in neuronal migration in lissencephaly patients. PMID- 15800060 TI - Differential roles for actin polymerization and a myosin II motor in assembly of the epithelial apical junctional complex. AB - Differentiation and polarization of epithelial cells depends on the formation of the apical junctional complex (AJC), which is composed of the tight junction (TJ) and the adherens junction (AJ). In this study, we investigated mechanisms of actin reorganization that drive the establishment of AJC. Using a calcium switch model, we observed that formation of the AJC in T84 intestinal epithelial cells began with the assembly of adherens-like junctions followed by the formation of TJs. Early adherens-like junctions and TJs readily incorporated exogenous G-actin and were disassembled by latrunculin B, thus indicating dependence on continuous actin polymerization. Both adherens-like junctions and TJs were enriched in actin related protein 3 and neuronal Wiskott-Aldrich syndrome protein (N-WASP), and their assembly was prevented by the N-WASP inhibitor wiskostatin. In contrast, the formation of TJs, but not adherens-like junctions, was accompanied by recruitment of myosin II and was blocked by inhibition of myosin II with blebbistatin. In addition, blebbistatin inhibited the ability of epithelial cells to establish a columnar phenotype with proper apico-basal polarity. These findings suggest that actin polymerization directly mediates recruitment and maintenance of AJ/TJ proteins at intercellular contacts, whereas myosin II regulates cell polarization and correct positioning of the AJC within the plasma membrane. PMID- 15800061 TI - Association of the leukocyte plasma membrane with the actin cytoskeleton through coiled coil-mediated trimeric coronin 1 molecules. AB - Coronin 1 is a member of the coronin protein family specifically expressed in leukocytes and accumulates at sites of rearrangements of the F-actin cytoskeleton. Here, we describe that coronin 1 molecules are coiled coil-mediated homotrimeric complexes, which associate with the plasma membrane and with the cytoskeleton via two distinct domains. Association with the cytoskeleton was mediated by trimerization of a stretch of positively charged residues within a linker region between the N-terminal, WD repeat-containing domain and the C terminal coiled coil. In contrast, neither the coiled coil nor the positively charged residues within the linker domain were required for plasma membrane binding, suggesting that the N-terminal, WD repeat-containing domain mediates membrane interaction. The capacity of coronin 1 to link the leukocyte cytoskeleton to the plasma membrane may serve to integrate outside-inside signaling with modulation of the cytoskeleton. PMID- 15800062 TI - Neutrophil migration across tight junctions is mediated by adhesive interactions between epithelial coxsackie and adenovirus receptor and a junctional adhesion molecule-like protein on neutrophils. AB - Neutrophil (polymorphonuclear leukocytes [PMN]) transepithelial migration during inflammatory episodes involves a complex series of adhesive interactions and signaling events. Previous studies have shown that key adhesive interactions between leukocyte CD11b/CD18 and basally expressed fucosylated glycoproteins followed by binding to desmosomal-associated JAM-C are key elements of the transmigration response. Here we provide the first evidence that PMN-expressed junctional adhesion molecule-like protein (JAML) regulates transmigration via binding interactions with epithelial coxsackie and adenovirus receptor (CAR). Experiments with a JAML fusion protein revealed specific binding of JAML to epithelial CAR expressed at tight junctions in T84 cell monolayers and normal human colonic mucosa. Furthermore, JAML-CAR binding is mediated via the membrane distal immunoglobulin (Ig) loop of CAR and the membrane proximal Ig loop of JAML. PMN bound to immobilized CAR but not JAML in a divalent cation-independent manner. Lastly, in assays of PMN transepithelial migration, JAML/CAR fusion proteins and their antibodies significantly inhibited transmigration in a specific manner. Taken together, these results indicate that JAML and CAR are a novel pair of adhesion molecules that play an important role in modulating PMN migration cross epithelial tight junctions. These findings add a new element to a multistep model of PMN transepithelial migration and may provide new targets for anti-inflammatory therapies. PMID- 15800063 TI - Galectin-3 is a downstream regulator of matrix metalloproteinase-9 function during endochondral bone formation. AB - Endochondral bone formation is characterized by the progressive replacement of a cartilage anlagen by bone at the growth plate with a tight balance between the rates of chondrocyte proliferation, differentiation, and cell death. Deficiency of matrix metalloproteinase-9 (MMP-9) leads to an accumulation of late hypertrophic chondrocytes. We found that galectin-3, an in vitro substrate of MMP 9, accumulates in the late hypertrophic chondrocytes and their surrounding extracellular matrix in the expanded hypertrophic cartilage zone. Treatment of wild-type embryonic metatarsals in culture with full-length galectin-3, but not galectin-3 cleaved by MMP-9, mimicked the embryonic phenotype of Mmp-9 null mice, with an increased hypertrophic zone and decreased osteoclast recruitment. These results indicate that extracellular galectin-3 could be an endogenous substrate of MMP-9 that acts downstream to regulate hypertrophic chondrocyte death and osteoclast recruitment during endochondral bone formation. Thus, the disruption of growth plate homeostasis in Mmp-9 null mice links galectin-3 and MMP-9 in the regulation of the clearance of late chondrocytes through regulation of their terminal differentiation. PMID- 15800064 TI - Mto2p, a novel fission yeast protein required for cytoplasmic microtubule organization and anchoring of the cytokinetic actin ring. AB - Microtubules regulate diverse cellular processes, including chromosome segregation, nuclear positioning, and cytokinesis. In many organisms, microtubule nucleation requires gamma-tubulin and associated proteins present at specific microtubule organizing centers (MTOCs). In fission yeast, interphase cytoplasmic microtubules originate from poorly characterized interphase MTOCs and spindle pole body (SPB), and during late anaphase from the equatorial MTOC (EMTOC). It has been previously shown that Mto1p (Mbo1p/Mod20p) function is important for the organization/nucleation of all cytoplasmic microtubules. Here, we show that Mto2p, a novel protein, interacts with Mto1p and is important for establishing a normal interphase cytoplasmic microtubule array. In addition, mto2Delta cells fail to establish a stable EMTOC and localize gamma-tubulin complex members to this medial structure. As predicted from these functions, Mto2p localizes to microtubules, the SPB, and the EMTOC in an Mto1p-dependent manner. mto2Delta cells fail to anchor the cytokinetic actin ring in the medial region of the cell and under conditions that mildly perturb actin structures, these rings unravel in mto2Delta cells. Our results suggest that the Mto2p and the EMTOC are critical for anchoring the cytokinetic actin ring to the medial region of the cell and for proper coordination of mitosis with cytokinesis. PMID- 15800065 TI - Quantitative SUMO-1 modification of a vaccinia virus protein is required for its specific localization and prevents its self-association. AB - Vaccinia virus (VV), the prototype member of the Poxviridae, a family of large DNA viruses, carries out DNA replication in specialized cytoplasmic sites that are enclosed by the rough endoplasmic reticulum (ER). We show that the VV gene product of A40R is quantitatively modified by SUMO-1, which is required for its localization to the ER-enclosed replication sites. Expression of A40R lacking SUMO-1 induced the formation of rod-shaped cytoplasmic aggregates. The latter likely consisted of polymers of nonsumoylated protein, because unmodified A40R interacted with itself, but not with the SUMO-1-conjugated protein. Using a bacterial sumoylation system, we furthermore show that unmodified A40R is mostly insoluble, whereas the modified form is completely soluble. By electron microscopy, the A40R rods seen in cells were associated with the cytosolic side of the ER and induced the apposition of several ER cisternae. A40R is the first example of a poxvirus protein to acquire SUMO-1. Its quantitative SUMO-1 modification is required for its proper localization to the viral "mini-nuclei" and prevents its self-association. The ability of the nonsumoylated A40R to bring ER membranes close together could suggest a role in the fusion of ER cisternae when these coalesce to enclose the VV replication sites. PMID- 15800066 TI - Control of Ste6 recycling by ubiquitination in the early endocytic pathway in yeast. AB - We present evidence that ubiquitination controls sorting of the ABC-transporter Ste6 in the early endocytic pathway. The intracellular distribution of Ste6 variants with reduced ubiquitination was examined. In contrast to wild-type Ste6, which was mainly localized to internal structures, these variants accumulated at the cell surface in a polar manner. When endocytic recycling was blocked by Ypt6 inactivation, the ubiquitination deficient variants were trapped inside the cell. This indicates that the polar distribution is maintained dynamically through endocytic recycling and localized exocytosis ("kinetic polarization"). Ste6 does not appear to recycle through late endosomes, because recycling was not blocked in class E vps (vacuolar protein sorting) mutants (Deltavps4, Deltavps27), which are affected in late endosome function and in the retromer mutant Deltavps35. Instead, recycling was partially affected in the sorting nexin mutant Deltasnx4, which serves as an indication that Ste6 recycles through early endosomes. Enhanced recycling of wild-type Ste6 was observed in class D vps mutants (Deltapep12, Deltavps8, and Deltavps21). The identification of putative recycling signals in Ste6 suggests that recycling is a signal-mediated process. Endocytic recycling and localized exocytosis could be important for Ste6 polarization during the mating process. PMID- 15800067 TI - Human ether-a-go-go-related gene 1 channels are physically linked to beta1 integrins and modulate adhesion-dependent signaling. AB - Adhesive receptors of the integrin family are primarily involved in cell extracellular matrix adhesion. Additionally, integrins trigger multiple signaling pathways that are involved in cell migration, proliferation, survival, and differentiation. We previously demonstrated that the activation of integrins containing the beta(1) subunit leads to a selective increase in potassium currents carried by the human ether-a-go-go-related gene (hERG) channels in neuroblastoma and leukemia cells; this current activation modulates adhesion dependent differentiation in these cells. We hypothesized that the cross-talk between integrins and hERG channels could be traced back to the assembly of a macromolecular signaling complex comprising the two proteins. We tested this hypothesis in both SH-SY5Y neuroblastoma cells and in human embryonic kidney 293 cells stably transfected with hERG1 and, therefore, expressing only the full length hERG1 protein on the plasma membrane. The beta(1) integrin and hERG1 coprecipitate in these cells and colocalize in both intracellular and surface membrane compartments. The two proteins also coprecipitate with caveolin-1, suggesting the localization of the complex in lipid rafts/caveolae. hERG1 transfected cells undergo an activation of hERG currents after beta(1) integrin mediated adhesion to fibronectin; concomitant with this activation, the focal adhesion kinase associates with the hERG1 protein and becomes tyrosine phosphorylated. Using hERG1-specific inhibitors, we show that the tyrosine phosphorylation of focal adhesion kinase is strictly dependent on hERG channel activity. Similarly, the activity of the small GTPase Rac1 turned out to be dependent on hERG currents. On the whole, these data indicate that the hERG1 protein associates with beta(1) integrins and modulates adhesion receptor signaling. PMID- 15800068 TI - Polarized gene expression determines woronin body formation at the leading edge of the fungal colony. AB - The Woronin body (WB) is a peroxisome-related organelle that is centered on a crystalline core of the HEX-1 protein, which functions to seal septal pores of filamentous ascomycetes in response to cellular damage. Here, we investigate the cellular and genetic control of WB-formation and show that polarized hex-1 gene expression determines WB-biogenesis at the growing hyphal apex. We find that intron splicing is coupled to efficient hex-1 gene expression and strikingly, when the yellow fluorescent protein was expressed from hex-1 regulatory sequences, we observed a fluorescent gradient that was maximal in apical cells. Moreover, endogenous hex-1 transcripts were specifically enriched at the leading edge of the fungal colony, whereas other transcripts accumulated in basal regions. Time-lapse confocal microscopy showed that HEX-1 crystals normally formed in the vicinity of the hyphal apex in large peroxisomes, which matured and were immobilized at the cell periphery as cells underwent septation. When the hex 1 structural gene was expressed from regulatory sequences of an abundant, basally localized transcript, WB-core formation was redetermined to basal regions of the colony, and these strains displayed loss-of-function phenotypes specifically in apical hyphal compartments. These results show that apically localized gene expression is a key determinant of spatially restricted WB-assembly. We suggest that this type of regulation may be widely used to determine cellular activity in apical regions of the fungal hypha. PMID- 15800070 TI - Corticotrophin-releasing factor augments the I(H) in rat hypothalamic paraventricular nucleus parvocellular neurons in vitro. AB - The goal of this study was to characterize the effects of corticotrophin releasing factor (CRF) on rat paraventricular nucleus (PVN) putative parvocellular neurons using whole cell patch-clamp recordings and single-cell reverse transcription-multiplex polymerase chain reaction (single-cell RT-mPCR) techniques. Under current clamp, CRF (10-600 nM) increased the neuronal basal firing rate and depolarized neurons in a dose-dependent manner. CRF-induced depolarization was unaffected by co-perfusion with TTX, 6-cyano-7 nitroquinoxaline-2 3-dione (CNQX), and bicuculline but was completely inhibited by ZD7288. Under voltage clamp, 300 nM CRF significantly increased the hyperpolarization-activated cation current (I(H)) in a voltage-dependent manner, shifted the I(H) conductance-voltage relationship (V 1/2) toward depolarization by approximately 7.8 mV, and enhanced the I(H) kinetics without changing the slope constant (k). Extracellular application of ZD7288 completely blocked I(H) and the CRF-induced increase in I(H). Furthermore, CRF-induced effects were completely blocked by extracellular application of 1 microM alpha-helical CRF-(9 14) (alpha-hCRF), a nonselective CRF receptor antagonist, but were not affected by extracellular application of antisauvagine-30, a selective CRF-receptor 2 antagonist. Single-cell RT-mPCR analysis showed that these neurons co-expressed CRF receptor 1 mRNA and CRF receptor 2 mRNA. Furthermore, CRF-sensitive neurons co-expressed HCN1 channel mRNA, HCN2 channel mRNA, and HCN3 channel mRNA, but not HCN4 channel mRNA. These results suggest that CRF modulates the subpopulation of PVN parvocellular neuronal function by CRF-receptor 1-mediated potentiation of HCN ion channel activity. PMID- 15800069 TI - Clues to CD2-associated protein involvement in cytokinesis. AB - Cytokinesis requires membrane trafficking coupled to actin remodeling and involves a number of trafficking molecules. CD2-associated protein (CD2AP) has been implicated in dynamic actin remodeling and membrane trafficking that occurs during endocytosis leading to the degradative pathway. In this study, we present several arguments for its implication in cytokinesis. First, endogenous CD2AP was found concentrated in the narrow region of the midzone microtubules during anaphase and in the midbody during late telophase. Moreover, we found that CD2AP is a membrane- and not a microtubule-associated protein. Second, the overexpression of the first two Src homology 3 domains of CD2AP, which are responsible for this localization, led to a significant increase in the rate of cell multinucleation. Third, the CD2AP small interfering RNA interfered with the cell separation, indicating that CD2AP is required for HeLa cells cytokinesis. Fourth, using the yeast two-hybrid system, we found that CD2AP interacted with anillin, a specific cleavage furrow component, and the two proteins colocalized at the midbody. Both CD2AP and anillin were found phosphorylated early in mitosis and also CD2AP phosphorylation was coupled to its delocalization from membrane to cytosol. All these observations led us to propose CD2AP as a new player in cytokinesis. PMID- 15800071 TI - Does a unique type of CA3 pyramidal cell in primates bypass the dentate gate? AB - The predominant excitatory synaptic input to the hippocampus arises from entorhinal cortical axons that synapse with dentate granule cells, which in turn synapse with CA3 pyramidal cells. Thus two highly excitable brain areas--the entorhinal cortex and the CA3 field--are separated by dentate granule cells, which have been proposed to function as a gate or filter. However, unlike rats, primates have "dentate" CA3 pyramidal cells with an apical dendrite that extends into the molecular layer of the dentate gyrus, where they could receive strong, monosynaptic, excitatory synaptic input from the entorhinal cortex. To test this possibility, the dentate gyrus molecular layer was stimulated while intracellular recordings were obtained from CA3 pyramidal cells in hippocampal slices from neurologically normal macaque monkeys. Stimulus intensity of the outer molecular layer of the dentate gyrus was standardized by the threshold intensity for evoking a dentate gyrus field potential population spike. Recorded proximal CA3 pyramidal cells were labeled with biocytin, processed with diaminobenzidine for visualization, and classified according to their dendritic morphology. In response to stimulation of the dentate gyrus molecular layer, action potential thresholds were similar in proximal CA3 pyramidal cells with different dendritic morphologies. These findings do not support the hypothesis that dentate CA3 pyramidal cells receive stronger synaptic input from the entorhinal cortex than do other proximal CA3 pyramidal cells. PMID- 15800072 TI - Adrenal chromaffin cells exhibit impaired granule trafficking in NCAM knockout mice. AB - Neural cell adhesion molecule (NCAM) plays several critical roles in neuron path finding and intercellular communication during development. In the clinical setting, serum NCAM levels are altered in both schizophrenic and autistic patients. NCAM knockout mice have been shown to exhibit deficits in neuronal functions including impaired hippocampal long term potentiation and motor coordination. Recent studies in NCAM null mice have indicated that synaptic vesicle trafficking and active zone targeting are impaired, resulting in periodic synaptic transmission failure under repetitive physiological stimulation. In this study, we tested whether NCAM plays a role in vesicle trafficking that is limited to the neuromuscular junction or whether it may also play a more general role in transmitter release from other cell systems. We tested catecholamine release from neuroendocrine chromaffin cells in the mouse adrenal tissue slice preparation. We utilize electrophysiological and electrochemical measures to assay granule recruitment and targeting in wild-type and NCAM -/- mice. Our data show that NCAM -/- mice exhibit deficits in normal granule trafficking between the readily releasable pool and the highly release-competent immediately releasable pool. This defect results in a decreased rate of granule fusion and thus catecholamine release under physiological stimulation. Our data indicate that NCAM plays a basic role in the transmitter release mechanism in neuroendocrine cells through mediation of granule recruitment and is not limited to the neuromuscular junction and central synapse active zones. PMID- 15800073 TI - Longitudinal depolarization gradients along the somatodendritic axis of CA1 pyramidal cells: a novel feature of spreading depression. AB - We studied the subcellular correlates of spreading depression (SD) in the CA1 rat hippocampus by combining intrasomatic and intradendritic recordings of pyramidal cells with extracellular DC and evoked field and unitary activity. The results demonstrate that during SD only specific parts of the dendritic membranes are deeply depolarized and electrically shunted. Somatic impalements yielded near zero membrane potential (V(m)) and maximum decrease of input resistance (R(in)) whether the accompanying extracellular negative potential (V(o)) moved along the basal, the apical or both dendritic arbors. However, apical intradendritic recordings showed a different course of local V(m) that is hardly detected from the soma. A decreasing depolarization gradient was observed from the edge of SD affected fully depolarized subcellular regions toward distal dendrites. Within apical dendrites, the depolarizing front moved toward and stopped at proximal dendrites during the time course of SD so that distal dendrites had repolarized in part or in full by the end of the wave. The drop of local R(in) was initially maximal at any somatodendritic loci and also recovered partially before the end of SD. This recovery was stronger and faster in far dendrites and is best explained by a wave-like somatopetal closure of membrane conductances. Cell subregions far from SD-affected membranes remain electrically excitable and show evoked unitary and field activity. We propose that neuronal depolarization during SD is caused by current flow through extended but discrete patches of shunted membranes driven by uneven longitudinal depolarization. PMID- 15800074 TI - Temperature affects voltage-sensitive conductances differentially in octopus cells of the mammalian cochlear nucleus. AB - Temperature is an important physiological variable the influence of which on macroscopic electrophysiological measurements in slices is not well documented. We show that each of three voltage-sensitive conductances of octopus cells of the mammalian ventral cochlear nucleus (VCN) is affected differently by changes in temperature. As expected, the kinetics of the currents were faster at higher than at lower temperature. Where they could be measured, time constants of activation, deactivation, and inactivation had Q10 values between 1.8 and 4.6. The magnitude of the peak conductances was differentially affected by temperature. While the peak magnitude of the high-voltage-activated K+ conductance, g(KH), was unaffected by changes in temperature, the peak of the low-voltage-activated K+ conductance, g(KL), was reduced by half when the temperature was lowered from 33 to 23 degrees C (Q10 = 2). Changing the temperature changed the kinetics and the magnitude of the hyperpolarization-activated mixed cation conductance, g(h), but the changes in magnitude were transient. The voltage sensitivity of the three conductances was unaffected by temperature. The action of temperature on these conductances is reflected in the resting potentials and in the shapes of action potentials. PMID- 15800075 TI - Control of locomotor cycle durations. AB - In intact animals and humans, increases in locomotor speed are usually associated with decreases in step cycle duration. Most data indicate that the locomotor central pattern generator (CPG) shortens cycle duration mainly by shortening the durations of extensor rather than flexor phases of the step cycle. Here we report that in fictive locomotion elicited by electrical stimulation of the midbrain locomotor region (MLR) in the cat, spontaneous variations in cycle duration were due more to changes in flexor rather than extensor phase durations in 22 of 31 experiments. The locomotor CPG is therefore not inherently extensor- or flexor biased. We coined the term "dominant" to designate the phase (flexion or extension) showing the larger variation. In a simple half-center oscillator model, experimental phase duration plots were fitted well by adjusting two parameters that corresponded to background drive ("bias") and sensitivity ("gain") of the oscillator's timing elements. By analogy we argue that variations in background drive to the neural timing elements of the CPG could produce larger variations in phase duration in the half-center receiving the lower background drive, i.e., background drive may determine which half-center is dominant. The fact that data from normal cats were also fitted well by the model indicates that sensory input and central drive combine to determine locomotor phase durations. We conclude that there is a considerable flexibility in the control of phase durations in MLR-induced fictive locomotion. We posit that this may be explained by changes in background excitation of neural timing elements in the locomotor CPG. PMID- 15800076 TI - Reducing contralateral SI activity reveals hindlimb receptive fields in the SI forelimb-stump representation of neonatally amputated rats. AB - In adult rats that sustained forelimb amputation on the day of birth, >30% of multiunit recording sites in the forelimb-stump representation of primary somatosensory cortex (SI) also respond to cutaneous hindlimb stimulation when cortical GABA(A+B) receptors are blocked (GRB). This study examined whether hindlimb receptive fields could also be revealed in forelimb-stump sites by reducing one known source of excitatory input to SI GABAergic neurons, the contralateral SI cortex. Corpus callosum projection neurons connect homotopic SI regions, making excitatory contacts onto pyramidal cells and interneurons. Thus in addition to providing monosynaptic excitation in SI, callosal fibers can produce disynaptic inhibition through excitatory synapses with inhibitory interneurons. Based on the latter of these connections, we hypothesized that inactivating the contralateral (intact) SI forelimb region would "unmask" normally suppressed hindlimb responses by reducing the activity of SI GABAergic neurons. The SI forelimb-stump representation was first mapped under normal conditions and then during GRB to identify stump/hindlimb responsive sites. After GRB had dissipated, the contralateral (intact) SI forelimb region was mapped and reversibly inactivated with injections of 4% lidocaine, and selected forelimb stump sites were retested. Contralateral SI inactivation revealed hindlimb responses in approximately 60% of sites that were stump/hindlimb responsive during GRB. These findings indicate that activity in the contralateral SI contributes to the suppression of reorganized hindlimb receptive fields in neonatally amputated rats. PMID- 15800077 TI - Reduction of common synaptic drive to ankle dorsiflexor motoneurons during walking in patients with spinal cord lesion. AB - It is possible to obtain information about the synaptic drive to motoneurons during walking by analyzing motor-unit coupling in the time and frequency domains. The purpose of the present study was to compare motor-unit coupling during walking in healthy subjects and patients with incomplete spinal cord lesion to obtain evidence of differences in the motoneuronal drive that result from the lesion. Such information is of importance for development of new strategies for gait restoration. Twenty patients with incomplete spinal cord lesion (SCL) participated in the study. Control experiments were performed in 11 healthy subjects. In all healthy subjects, short-term synchronization was evident in the discharge of tibialis anterior (TA) motor units during the swing phase of treadmill walking. This was identified from the presence of a narrow central peak in cumulant densities constructed from paired EMG recordings and from the presence of significant coherence between these signals in the 10- to 20-Hz band. Such indicators of short-term synchrony were either absent or very small in the patient group. The relationship between the amount of short-term synchrony and the magnitude of the 10- to 20-Hz coherence in the patients is discussed in relation to gait ability. It is suggested that supraspinal drive to the spinal cord is responsible for short-term synchrony and coherence in the 10- to 20-Hz frequency band during walking in healthy subjects. Absence or reduction of these features may serve as physiological markers of impaired supraspinal control of gait in SCL patients. Such markers could have diagnostic and prognostic value in relation to the recovery of locomotion in patients with central motor lesions. PMID- 15800078 TI - Whole cell stochastic model reproduces the irregularities found in the membrane potential of bursting neurons. AB - Irregular intrinsic behavior of neurons seems ubiquitous in the nervous system. Even in circuits specialized to provide periodic and reliable patterns to control the repetitive activity of muscles, such as the pyloric central pattern generator (CPG) of the crustacean stomatogastric ganglion (STG), many bursting motor neurons present irregular activity when deprived from synaptic inputs. Moreover, many authors attribute to these irregularities the role of providing flexibility and adaptation capabilities to oscillatory neural networks such as CPGs. These irregular behaviors, related to nonlinear and chaotic properties of the cells, pose serious challenges to developing deterministic Hodgkin-Huxley-type (HH-type) conductance models. Only a few deterministic HH-type models based on experimental conductance values were able to show such nonlinear properties, but most of these models are based on slow oscillatory dynamics of the cytosolic calcium concentration that were never found experimentally in STG neurons. Based on an up to-date single-compartment deterministic HH-type model of a STG neuron, we developed a stochastic HH-type model based on the microscopic Markovian states that an ion channel can achieve. We used tools from nonlinear analysis to show that the stochastic model is able to express the same kind of irregularities, sensitivity to initial conditions, and low dimensional dynamics found in the neurons isolated from the STG. Without including any nonrealistic dynamics in our whole cell stochastic model, we show that the nontrivial dynamics of the membrane potential naturally emerge from the interplay between the microscopic probabilistic character of the ion channels and the nonlinear interactions among these elements. Moreover, the experimental irregular behavior is reproduced by the stochastic model for the same parameters for which the membrane potential of the original deterministic model exhibits periodic oscillations. PMID- 15800079 TI - Dissection of bidirectional synaptic plasticity into saturable unidirectional processes. AB - In populations of synapses, overall synaptic strength can undergo either a net strengthening (long-term potentiation) or weakening (long-term depression). These phenomena have distinct induction pathways, but the functional outcome is usually measured as a single lumped quantity. In hippocampal CA3-CA1 synapses, we took two approaches to study the activity dependence of each phenomenon in isolation. First, we selectively blocked one process by applying kinase or phosphatase inhibitors known, respectively, to block potentiation or depression. Second, we saturated depression or potentiation and examined the activity dependence of the converse process. The resulting unidirectional learning rules could be recombined to give a well-known bidirectional frequency-dependent learning rule under the assumption that when both pathways are activated kinases dominate, resulting in potentiation. Saturation experiments revealed an additional process in which potentiated synapses can be locked at high strength. Saturability of the components of plasticity implies that the amount of plasticity contributed by each pathway depends on the initial level of strength of the synapses. Variation in the distribution of initial synaptic strengths predicts a form of metaplasticity and can account for differences in learning rules observed under several physiological and genetic manipulations. PMID- 15800080 TI - Whither "chronic prostatitis"? PMID- 15800081 TI - Lymphogranuloma venereum in the United Kingdom. PMID- 15800082 TI - Foundation training and genitourinary medicine. PMID- 15800083 TI - Life in the littoral zone: lactobacilli losing the plot. AB - Recurrent bacterial vaginosis is a challenge for those affected by it, and their physicians. Our inability to prevent relapse after treatment, may be because of the flawed approach of using antibiotics to treat a condition that is an imbalance rather than an infection. The maintenance of a healthy lactobacillus population offers an approach to preventing relapse: the problem is how best to do this. Physiological approaches such as the use of hydrogen peroxide, lactic acid, and exogenous lactobacilli need to be explored further. The role of bacterial vaginosis as a risk factor for acquisition of HIV and other STIs is a further impetus to attempting to prevent bacterial vaginosis from recurring. PMID- 15800084 TI - A systematic review of the epidemiology and interaction of herpes simplex virus types 1 and 2. AB - OBJECTIVES: To explore epidemiological evidence about the interaction of herpes simplex virus (HSV) 1 and HSV-2 infections. METHODS: A systematic review was undertaken of published epidemiological studies describing the pattern of HSV-1 or HSV-2 by age, and the coincidence of the two viral infections. RESULTS: In cross sectional studies the unadjusted odds of HSV-2 are greater in those with HSV-1 infection in study populations categorised as "low risk" (p = 0.06) and across European populations (p = 0.001). This was not evident in "high risk" populations or in the United States. This increased risk of HSV-2 in those with HSV-1 infection does not agree with the results of prospective studies where there is a non-significant trend towards a lower risk of HSV-2 infection associated with previous HSV-1 infection. CONCLUSIONS: "Low risk" and European populations have a relatively low HSV-2 seroprevalence and infection is more concentrated in those with characteristics putting them at high risk for both HSV 1 and HSV-2. This confounding could mask any protective effect of HSV-1, which is hinted at, but not demonstrated, in prospective and adjusted studies. PMID- 15800085 TI - Attitudes to HIV and HIV testing in high prevalence areas of China: informing the introduction of voluntary counselling and testing programmes. AB - OBJECTIVES: To assess knowledge and attitudes towards HIV and its testing among pregnant women and health professionals in Yunnan Province, south west China, to inform the introduction of voluntary counselling and testing (VCT) programmes. METHODS: The study design was a cross sectional survey using self completion questionnaires. It was carried out in 12 hospitals in four high prevalence areas of Yunnan Province. Questionnaires were completed under examination conditions by health professionals, and at the routine antenatal examination by pregnant women. RESULTS: Completed questionnaires were obtained from 840 pregnant women and 780 health professionals. Knowledge of HIV and its modes of transmission were good in health professionals but patchy in pregnant women. The weakest area in both groups was knowledge of maternal to child transmission. There was strong support for compulsory testing in pregnancy and at the premarital examination. But attitudes towards HIV/AIDS were negative: 23% of health professionals and 45% of pregnant women thought HIV was a disease of "low class and illegal" people, 48% of health professionals and 59% of pregnant women thought that HIV positive individuals should not be allowed to get married, and 30% of the health professionals were not willing to treat an HIV positive individual. Levels of knowledge were higher and attitudes more positive in younger health professionals and better educated pregnant women. CONCLUSIONS: Community education programmes and intensive training of health workers must precede or accompany VCT programmes. They must particularly address negative attitudes towards people with HIV. Pilot VCT programmes are now under way in two of the areas studied. PMID- 15800086 TI - Discriminatory attitudes towards people living with HIV/AIDS and associated factors: a population based study in the Chinese general population. AB - OBJECTIVES: To examine the level of discriminatory attitudes towards people living with HIV/AIDS (PLWHA) and factors in association with such attitudes. METHODS: A population based cross sectional telephone survey was conducted. A total of 808 Hong Kong Chinese aged 18-50 randomly selected from the general population participated in the study. RESULTS: Around 42% of the respondents exhibited discriminatory attitudes in at least five out of the 20 relevant items. For instance, about 42% would avoid making physical contact with PLWHA; 35% believed that all infected medical staff should be dismissed and about 47% would agree with enacting a law to prohibit PLWHA from visiting Hong Kong. A sizeable proportion of the respondents also hold negative perceptions about PLWHA (for example, 43.7% agreed that the majority of PLWHA are promiscuous, 20.7% thought that PLWHA are merely receiving the punishment they deserve, etc). Multiple regression analysis found that age, HIV related knowledge, the above mentioned negative perceptions about PLWHA, fear related to AIDS, and exposure to HIV related information were independent predictors of discriminatory attitudes towards PLWHA. About 30% would give PLWHA the lowest priority in resource allocation among five groups of patients with chronic diseases. CONCLUSIONS: The general public in Hong Kong has formed some negative perceptions of PLWHA. Discriminatory attitudes towards PLWHA are common and cover different aspects of their life. Intervention programmes are warranted and an integrated approach is required. PMID- 15800087 TI - The effects of urethritis on seminal plasma HIV-1 RNA loads in homosexual men not receiving antiretroviral therapy. AB - OBJECTIVES: To examine the effects of urethritis and its treatment on semen plasma HIV-1 RNA load in HIV-1 infected men not receiving antiretroviral therapy (ART), in a developed world setting. METHODS: Prospective case-control study. HIV 1 infected homosexual men, not receiving ART for at least 3 months, with (cases) and without (controls) symptomatic urethritis, were recruited. Blood and semen were collected for HIV-1 RNA quantification at presentation, before antibiotic therapy, and at 1 and 2 weeks. RESULTS: 20 cases (13 gonococcal urethritis and/or chlamydial urethritis (GU/CU) and seven non-specific urethritis (NSU)) and 35 controls were recruited. Baseline characteristics and blood plasma viral load were similar in cases and controls. Mean log semen plasma viral loads were higher among those with GU/CU compared with controls (4.27 log versus 3.55 log respectively; p = 0.01) but not in those with NSU (3.48 log; p = 0.82). Following antibiotics, semen plasma viral loads fell by a mean of 0.25 log (95% CI: 0.03 to 0.47) in those with GU/CU. Semen plasma viral loads did not fall in those with NSU. CONCLUSIONS: In this study of 55 homosexual men not on ART, semen plasma viral loads were approximately fivefold higher in those with GU/CU, but not NSU, compared with controls. Treatment of GU/CU resulted in reduction in semen plasma viral loads. Although absolute effects were considerably lower when compared to patients from a similar study from sub-Saharan Africa, our data demonstrate the potential for sexually transmitted infections to enhance HIV infectivity of men not receiving ART in the developed world. PMID- 15800088 TI - Incorporating a social networking approach to enhance contact tracing in a heterosexual outbreak of syphilis. AB - OBJECTIVE: This report describes the outcomes of a social networking approach (SNA) in a heterosexually transmitted syphilis epidemic by street nurses in British Columbia, Canada. METHODS: Street nurses used SNA interviewing cues, environmental observation, peer outreach, serial interviewing, conducted blood tests, and offered treatment on the street. Rates of cases of syphilis identified by outreach nurses and cases linked to another case were compared before and after adoption of an SNA. RESULTS: SNA resulted in a significantly increased proportion of cases identified by the street nurses (p = 0.01) and increased the percentage of cases linked to a previous case (p = 0.03). CONCLUSION: This preliminary study confirms that SNA can increase the number of cases and contacts identified in an epidemic of a sexually transmitted disease in a vulnerable, hard to reach population. PMID- 15800090 TI - Early syphilis presenting as a painful polyradiculopathy in an HIV positive individual. AB - Neurosyphilis and neurological complications from syphilis may be commoner in HIV disease. With outbreaks of early syphilis in HIV positive individuals being observed over recent years, rare neurological manifestations of secondary syphilis will be observed more commonly. We describe a case of an HIV positive individual whose first presenting feature of early syphilis was a polyradiculopathy. PMID- 15800089 TI - Biological and hormonal markers of chlamydia, human papillomavirus, and bacterial vaginosis among adolescents attending genitourinary medicine clinics. AB - OBJECTIVE: To assess maturity indices, menstrual patterns, hormonal factors, and risk of adolescent genital tract infections. METHODS: Cross sectional study in three genitourinary medicine clinics. Females 17 years or less, within 5 years of menarche, or reporting oligo-amenorrhoea were screened for genital tract infections and menstrual cycle characteristics determined. The outcome measures were risk factors associated with chlamydia, human papillomavirus (HPV DNA) and bacterial vaginosis (BV), separately and pooled. Correlations between estrone-3 glucuronide (E3G) and pregnanediol-3alpha-glucuronide (P3G) hormone concentrations and chlamydia, HPV, and BV. RESULTS: Among 127 adolescents, HPV was present in 64.4% (95% CI: 54.5 to 74.3), BV in 33.9% (19.1 to 34.5), and chlamydia in 26.8% (19.1 to 34.5). Breast maturity, oligomenorrhoea, and older gynaecological age were associated with lower risk of all infections. After adjustment for calendar age, race, and behavioural factors, gynaecological age remained significant (OR = 0.7, 0.6-0.9; p = 0.008). Behavioural risk factors differed by infection. Smoking was protective for HPV (OR = 0.1, 0.0 to 0.9; p = 0.007), and a recent new partner for chlamydia (OR = 0.3, 0.1 to 0.9; p = 0.024). Sex during menses was associated with increased BV risk (OR = 3.3, 1.5 to 7.2; p = 0.003). Chlamydia was higher among adolescents who used emergency contraception (2.5; 1.1 to 5.9, p = 0.029) and lower among those using condoms at last sex (OR = 0.3, 0.1 to 0.9; p = 0.015). Among 25 adolescents not using hormonal contraceptives, 15 had disturbed or anovulatory cycles. Chlamydia risk was inversely associated with P3G concentrations (Mann-Whitney; p = 0.05). CONCLUSIONS: Adolescents engaging in high risk behaviour at a young gynaecological age are susceptible to multiple infections. Adolescent clinical assessment should include gynaecological age. PMID- 15800091 TI - Mexican physicians' knowledge and attitudes about the human papillomavirus and cervical cancer: a national survey. AB - OBJECTIVE: To assess Mexican physicians' knowledge about the human papillomavirus (HPV) and cervical cancer and their opinions and practices related to screening, managing, and counselling women on these topics. METHODOLOGY: In August 2002 we surveyed 1206 general practitioners (GPs) and obstetricians-gynaecologists (Ob Gyns) working in a nationally representative sample of public and private facilities in urban Mexico. Eligible physicians completed a self administered questionnaire. We conducted a weighted analysis and used chi(2) tests to compare GPs and Ob-Gyns on outcome variables. RESULTS: 76% of recruited physicians responded to the survey. 43% of Ob-Gyns had performed a hysterectomy in the last year to treat a case of CIN I or II. With respect to HPV, while 80% of respondents identified the virus as the principal cause of cervical cancer, many lacked detailed knowledge about this association. Ob-Gyns were more likely than GPs to have heard about specific oncogenic strains of HPV (p<0.001). Nearly all respondents thought that women should be informed that HPV causes cervical cancer; nevertheless, physicians believed that positioning cervical cancer as a sexually transmitted infection (STI) could cause problems in partner relationships (60%), confusion (40%), and unnecessary anxiety among women (32%). CONCLUSIONS: Mexican physicians support patient education on the HPV-cervical cancer link. However, findings suggest the need to present clear messages to women (emphasising, for example, that only certain types of HPV are oncogenic), to consider the conflicts such information might create for couples, and to further educate physicians about this topic and about overall cervical cancer screening and treatment protocols. PMID- 15800092 TI - The value of anal cytology and human papillomavirus typing in the detection of anal intraepithelial neoplasia: a review of cases from an anoscopy clinic. AB - BACKGROUND: Previous studies have reached differing conclusions about the utility of anal cytology as a screening tool for anal intraepithelial neoplasia (AIN). There is a need also to establish whether HPV typing offers a useful adjunct to screening. METHODS: We analysed data from 99 consecutive homosexual/bisexual male patients (89 HIV-1 positive) who underwent high resolution anoscopy. Follow up visits for these patients were also included, giving a total of 160 anoscopic procedures. Comparison was made between results of anal cytology using the sampling method of Palefsky, and histological findings of biopsies taken from abnormal areas seen on high resolution anoscopic examination of the anal canal. Swabs taken concurrently with the cytology were analysed for the presence of human papillomavirus (HPV) DNA and compared with the cytological and histological findings. RESULTS: The sensitivity of the cytology was 83%, and the specificity 38% when compared with histology. At screening of 34 asymptomatic men, 83% had anal cytological dysplasia and 78% had AIN. There were no significant differences in the prevalence of hrHPV genotypes between different cytological or histological grades of abnormalities. CONCLUSION: Anal cytology by the Palefsky method is simple to undertake, has a sensitivity and specificity comparable with cervical cytology, and can therefore be used as the basis of a pilot screening project in centres with large cohorts of HIV positive homosexual men who have a high risk of developing anal carcinoma. HPV genotyping is not a useful adjunct to cytological screening. PMID- 15800093 TI - The response to sertraline in men with chronic pelvic pain syndrome. AB - OBJECTIVES: Male chronic pelvic pain syndrome (CPPS) is difficult to manage. Although antidepressants are frequently used in clinical practice, to date no interventional study has been published. We investigated men with CPPS to assess their response to the serotonin specific reuptake inhibitor (SSRI) antidepressant, sertraline. METHODS: Men with CPPS underwent a four glass test to exclude an infective cause for their symptoms. They were randomised to sertraline or matched placebo for 13 weeks after which they were unblinded. They were then allowed to either continue sertraline or cross over to active treatment for a further 13 weeks. Prostatic symptom severity (PSS) and frequency (PSF) scores, the Hospital Anxiety and Depression (HAD) scale and a psychosexual (PSex) questionnaire were completed at 0, 6, 13, and 26 weeks. Statistical analysis was by the Mann-Whitney U and Wilcoxon signed rank tests. RESULTS: 14 men enrolled. At week 13 there was a mean reduction in PSS scores of 6.1 in the active and 2.0 in placebo group, and in PSF scores of 3.6 and 1.0, respectively. There was no statistically significant difference in the PSS and PSF scores between the active versus placebo group because of the small subject numbers. If analysed as a case series, there was a significant reduction in PSS (11.7; p = 0.01) and PSF (5.9; p = 0.03) from baseline following 13 weeks of sertraline. There was also a decrease in mean HAD depression score from 4.6 at baseline to 2.4. CONCLUSION: Sertraline led to a significant improvement in prostatic symptom severity and frequency from baseline following 13 weeks of treatment. Although this analysis does not exclude a placebo effect, the randomised placebo controlled findings show a trend to improvement with sertraline when compared to placebo. PMID- 15800094 TI - Patient, provider, and clinic characteristics associated with public STD clinic patient satisfaction. AB - OBJECTIVES: There is a lack of information describing levels of patient satisfaction among patients seeking sexually transmitted diseases (STD) care in a public clinic setting. We sought to identify patient, provider, and clinic characteristics associated with patient satisfaction within public STD clinics. METHODS: A cross sectional survey with random sampling was conducted among patients attending two public STD clinics. Satisfaction was assessed using questions from validated national surveys. Outcomes for multivariate logistic regression analysis were ratings of overall health care and clinician. RESULTS: 499/605 (82%) patients were enrolled. Patients were mean age 29 years, 51% male, 94% black. Lower rating of clinician technical skills (OR = 15.6 clinic A, OR = 7.9 clinic B) and clinic environment (OR = 3.9 clinic A, OR = 9.6 clinic B) were associated with lower healthcare rating, as was lower rating of television/video in waiting room (OR = 10.2, clinic A) and dysuria (OR = 4.2, clinic B). Higher clinician rating (OR = 0.58, clinic A) and receiving written materials (OR = 0.44, clinic B) were protective of lower healthcare rating. Risks for lower clinician rating at clinic A were greater pain, problems getting care, lower rating of clinician technical skill, and overall health care, while receiving written materials was protective. At clinic B, lower rating of clinician technical skill and clinic environment were risks for lower clinician rating. CONCLUSIONS: Patient satisfaction was associated with modifiable provider and clinic characteristics. Results from our study indicate a need to examine whether health outcomes of STD management vary by patient satisfaction. PMID- 15800095 TI - Methods employed by genitourinary medicine clinics in the United Kingdom to diagnose bacterial vaginosis. AB - OBJECTIVE: To determine the methods used by genitourinary medicine (GUM) clinics in the United Kingdom for the diagnosis of bacterial vaginosis (BV). METHODS: A questionnaire survey of UK GUM clinics was conducted. RESULTS: 148/221 (67%) clinics returned a questionnaire. 96/148 (64.9%) clinics reported using Amsel's criteria to diagnose BV but only 29 (30.5%) of these used all four of the composite criteria. 139/148 (93.9%) clinics used the appearance of a Gram stained vaginal smear as an aid in BV diagnosis, although a variety of scoring methods was employed. In the majority of clinics, 92/148 (62.2%), one staff discipline provided the microscopy service, in 50 (33.8%) clinics two staff disciplines provided microscopy services. The bulk of microscopy services within UK GUM clinics is provided by nurses. CONCLUSIONS: Most UK GUM clinics utilise the appearance of a Gram stained vaginal smear for the diagnosis of BV although there is little consensus at present about the type of scoring method employed. Adaptation of a uniform scoring method would have enormous benefits, including consistency and reproducibility of results and the development of quality assurance schemes for BV diagnosis on a national basis. There are important issues to be addressed regarding the initial training and ongoing support for nurses providing microscopy services within UK GUM clinics. PMID- 15800096 TI - "We don't really have cause to discuss these things, they don't affect us": a collaborative model for developing culturally appropriate sexual health services with the Bangladeshi community of Tower Hamlets. AB - OBJECTIVE: To identify barriers to accessing sexual health care among the Bangladeshi community of east London and to develop a model of community participation in service development. METHODS: Qualitative study using one to one interviews with sexual health service users plus focus groups in community settings. RESULTS: 58 people participated in the study, 12 in individual interviews and the remainder in six focus groups. All were of Bangladeshi origin. Four main themes were reported as impacting on access to services; confidentiality concerns, relevance of services to the community, problems with discussing sexual issues, and problems with previous experiences of health promotion. Community values regarding sex outside of marriage were an important underlying factor in participants' responses. Existing sexual health services were seen as culturally insensitive by patients and community groups. CONCLUSIONS: Community based health initiatives among hard to reach ethnic minority groups should use existing networks of statutory and non-statutory groups to benefit from local expertise and relationships. Steering groups composed of members of the local communities served by the clinic can usefully inform service development. PMID- 15800097 TI - Symptoms of non-gonococcal urethritis in heterosexual men: a case control study. AB - OBJECTIVE: To determine microbial and behavioural factors contributing to non gonococcal urethral symptoms in men. METHODS: Case-control study of heterosexual men with non-gonococcal urethral symptoms (cases) and without urethral symptoms (controls) attending Melbourne Sexual Health Centre, Australia. Sexual behaviour and condom use were measured by questionnaire. First stream urine was tested for potential pathogens: Chlamydia trachomatis (ligase chain reaction), Mycoplasma genitalium (polymerase chain reaction, PCR), Ureaplasma urealyticum (culture and PCR), and Streptococcus spp, Gardnerella vaginalis, and Haemophilus species (culture). Urethral smears from cases were examined for polymorphonuclear leucocytes. RESULTS: 80 cases and 79 controls were recruited over 4 months in 2002-3. 49 cases (61%) had urethritis by microscopic criteria, 17 (21%) had Chlamydia trachomatis (adjusted odds ratio (OR) 27 (95% confidence interval (CI): 3.4 to 222)), five (6%) had Mycoplasma genitalium (OR 6.1 (95% CI: 0.6 to 61)), and 11 (14%) had Gardnerella vaginalis (OR 9.0 (95% CI: 1.6 to 52)). Other organisms were not significantly associated with urethral symptoms. The presence of urethritis on urethral smear did not predict the presence of Chlamydia trachomatis (OR 1.7 (95% CI: 0.5 to 5.4)). Urethral symptoms were significantly associated with unprotected vaginal sex with more than one casual partner (OR 9.3 (95% CI: 1.3 to 65)) and unprotected anal sex with a regular partner in the past month (OR 3.5 (95% CI: 1.0 to 13)). CONCLUSION: Gardnerella vaginalis and unprotected anal sex may cause symptoms of non-gonococcal urethritis. Microscopy of the urethral smear to diagnose urethritis in this population does not help to identify which men with urethral symptoms require treatment for chlamydia. PMID- 15800098 TI - Results of a randomised trial of male condom promotion among Madagascar sex workers. AB - OBJECTIVES: To test the effect of supplementing peer promotion of male condom use with clinic based counselling, measured in terms of STI prevalence and reported male condom use. METHODS: 1000 female sex workers in Madagascar were randomised to two study arms: peer education supplemented by individual risk reduction counselling by a clinician (peer + clinic) versus condom promotion by peer educators only (peer only). STI testing was conducted at baseline and 6 months. Behavioural interviews were administered at baseline, 2, 4, and 6 months. RESULTS: At baseline, women in the peer only arm had prevalences of 16.0%, 23.6%, and 12.1% for chlamydia, gonorrhoea, and trichomoniasis respectively, with an aggregate prevalence of 38.2%. Baseline STI prevalences for the peer + clinic arm were slightly lower and 34.1% in aggregate. At 6 months, aggregate STI prevalence increased in the peer only arm to 41.4%, whereas the aggregate prevalence diminished slightly to 32.1% in the peer + clinic arm. In logistic regression analyses, the estimated odds ratios (ORs) for chlamydia, gonorrhoea, trichomoniasis, and aggregate STI were 0.7 (95% confidence interval 0.4 to 1.0), 0.7 (0.5 to 1.0), 0.8 (0.6 to 1.2), and 0.7 (0.5 to 0.9) respectively, comparing the peer + clinic arm with the peer only arm. The logistic regression OR for reported condom use with clients in the past 30 days increased from 1.1 at 2 months to 1.8 at 6 months, comparing the peer + clinic arm with the peer only arm, and was 1.4 overall (1.1 to 1.8). Adjustment for baseline factors changed the regression results little. CONCLUSIONS: The impact of male condom promotion on behaviour can be heightened through more concentrated counselling on risk reduction. Persistently high STI prevalence despite increases in reported condom use by sex workers supports the need for multidimensional control programmes. PMID- 15800100 TI - Female sex workers and fear of stigmatisation. PMID- 15800099 TI - Building a sentinel surveillance system for sexually transmitted infections in Germany, 2003. AB - BACKGROUND/OBJECTIVES: Increases in STIs have been reported from the United States and Europe. Since 2001, only syphilis and HIV are notifiable in Germany. A sentinel surveillance system has been set up to assess the occurrence and trends of STIs and identify risk groups. METHODS: Through the sentinel system data are collected from local health offices (LHO), hospital based STI clinics and private practitioners (dermato-venerology, urology, gynaecology, or HIV). For every newly diagnosed laboratory confirmed infection of HIV, gonorrhoea, chlamydia, syphilis, or trichomoniasis physicians complete a standardised questionnaire regarding diagnosis, source of infection, and demographic information. Patients complete a questionnaire about sexual risk behaviour. The patient form is matched with the diagnosis form using a unique identifier number. Characteristics of sentinel patients were compared with those reported through the HIV and syphilis national notification system. RESULTS: 58 LHO, 14 hospital based STI clinics, and 160 private practitioners (53.1% dermato-venerologists) from all federal states participated in the study. 176 (75.9%) sentinel sites are located in cities of >100 000 inhabitants. From 1 March 2003-29 February 2004, a total of 1833 STIs have been reported, among them 452 chlamydia, 321 syphilis (10.9% of notified syphilis), 343 gonorrhoea, 269 HIV (15.7% of notified HIV). 925 (50.5%) of the patients were male, the median age was 31 years. Female patients were more often of foreign origin (chi(2) test; 70.0% v 26.3%; p<0.001). CONCLUSIONS: Our sentinel system will provide a base for detection of STI trends in Germany. In addition, information about sexual risk behaviour will enable us to target prevention at those most at risk for STIs. PMID- 15800101 TI - Hepatitis B and C viral infections among STD clinic patients in India. PMID- 15800102 TI - A video mobile phone and herpes simplex. PMID- 15800103 TI - Clinically resistant trichomoniasis. PMID- 15800104 TI - Non-disclosure of previously known HIV seropositivity in patients "newly" diagnosed with HIV infection. PMID- 15800105 TI - Molluscum contagiosum presenting as penile horn in an HIV positive patient. PMID- 15800106 TI - Atypical presentation of lobar nephronia in an adult co-infected with HIV and hepatitis C. PMID- 15800107 TI - Contraception's proved potential to fight HIV. PMID- 15800108 TI - Collecting the essence of man: semen collection for HIV transmission studies in sub-Saharan Africa. PMID- 15800109 TI - Distribution and risk factors of hepatitis B, hepatitis C, and HIV infection in a female population with "illegal social behaviour". PMID- 15800110 TI - First case of spectinomycin resistant Neisseria gonorrhoeae isolate in New Delhi, India. PMID- 15800111 TI - High HIV risk profile among female commercial sex workers in Vinnitsa, Ukraine. PMID- 15800112 TI - Are all genital Chlamydia trachomatis infections pathogenic? PMID- 15800113 TI - Contrasting roles of complement activation and its regulation in membranous nephropathy. AB - The complement system is involved in defense against microorganisms, the processing of immune complexes and apoptotic debris, and the development of an appropriate immune response. Along with these physiologic effects, complement activation has the potential to result in tissue pathology. To limit this, various complement regulatory proteins (CRP) are present on host cells, including the glomerular podocyte. Experimental data from the Heymann nephritis (HN) rat model of human membranous nephropathy (MN) have shown that IgG antibodies in subepithelial immune deposits initiate complement activation and C5b-9-mediated damage of the overlying podocyte. Although IgG can activate the classical pathway, there also is evidence that alternative pathway activation occurs in MN, which could occur because of absent, dysfunctional, or inhibited podocyte CRP. Related to this are experimental data in HN showing the presence of antibodies that bind and inhibit podocyte CRP; although such antibodies have not been documented in human MN, a decrease in CR1 quantity on the podocyte has been observed. A s a result of a relative lack of CRP and the exposure of activating complement proteins to tubular cells, alternative complement pathway activation and C5b-9-mediated tubular injury can occur in MN and other proteinuric diseases. Overall, in a disease such as MN, the balance between complement regulation and activation is tipped toward its being activated. Therefore, a number of therapeutic approaches have been developed to counteract this, including recombinant forms of endogenous CRP and complement-inhibitory monoclonal antibodies. There is good reason to be optimistic that approaches to block complement activation will become viable therapy for human MN in the future. PMID- 15800114 TI - Membranous nephropathy: a long road but well traveled. PMID- 15800115 TI - Complement factor H mutation in familial thrombotic thrombocytopenic purpura with ADAMTS13 deficiency and renal involvement. AB - Thrombotic thrombocytopenic purpura is a rare disorder of small vessels that is associated with deficiency of the von Willebrand factor-cleaving protease ADAMTS13, which favors platelet adhesion and aggregation in the microcirculation. The disease manifests mainly with central nervous system symptoms, but cases of renal insufficiency have been reported. Presented are findings of the genetic basis of phenotype heterogeneity in thrombotic thrombocytopenic purpura in two sisters within one family. The patients had ADAMTS13 deficiency as a result of two heterozygous mutations (causing V88M and G1239V changes). In addition, a heterozygous mutation (causing an S890I change) in factor H of complement was found in the patient who developed chronic renal failure but not in her sister, who presented with exclusive neurologic symptoms. PMID- 15800116 TI - Membranoproliferative glomerulonephritis type II (dense deposit disease): an update. AB - Membranoproliferative glomerulonephritis type II (MPGN II) is a rare disease characterized by the deposition of abnormal electron-dense material within the glomerular basement membrane of the kidney and often within Bruch's membrane in the eye. The diagnosis is made in most patients between the ages of 5 and 15 yr, and within 10 yr, approximately half progress to end-stage renal disease, occasionally with the late comorbidity of visual impairment. The pathophysiologic basis of MPGN II is associated with the uncontrolled systemic activation of the alternative pathway (AP) of the complement cascade. In most patients, loss of complement regulation is caused by C3 nephritic factor, an autoantibody directed against the C3 convertase of the AP, but in some patients, mutations in the factor H gene have been identified. For the latter patients, plasma replacement therapy prevents renal failure, but for the majority of patients, there is no proven effective treatment. The disease recurs in virtually all renal allografts, and a high percentage of these ultimately fail. The development of molecular diagnostic tools and new therapies directed at controlling the AP of the complement cascade either locally in the kidney or at the systemic level may lead to effective treatments for MPGN II. PMID- 15800117 TI - Management of membranous nephropathy: when and what for treatment. PMID- 15800118 TI - Serum creatinine as marker of kidney function in South Asians: a study of reduced GFR in adults in Pakistan. AB - Migrant populations of South Asian origin have a higher risk for chronic kidney disease than the native whites. Several formulas have been developed to estimate kidney function from serum creatinine concentration. However, none of these has been validated in the South Asian population, which generally has different muscle mass composition than whites. A population-based cross-sectional study was performed on 262 individuals who were aged > or = 40 yr in Karachi, Pakistan. Reduced GFR was defined as creatinine clearance (Ccr) measured in 24-h urine collection of <60 ml/min per 1.73 m2. Creatinine excretion was compared with age- and gender-matched white individuals by comparison of observed versus expected results on the basis of a formula using t test. The agreement among Cockcroft Gault (CG) Ccr and Modification of Diet in Renal Disease (MDRD) Study GFR equations was assessed by regression analyses, and the degree of accuracy of estimated versus measured GFR was determined. Mean (95% confidence interval) creatinine excretion was 1.7 (1.0 to 2.4) mg/kg per d lower than expected for age and gender-matched white individuals (P < 0.001). The coefficient of determination for measured Ccr on the logarithmic scale was 66.7 and 55.6% for the CG and MDRD Study equations, respectively. The proportion of estimates within 20, 30, and 50% of measured Ccr values was 47.7 versus 32.8% (P < 0.001), 64.9 versus 49.6% (P < 0.001), and 79.4 versus 72.9 (P = 0.07) for CG versus MDRD Study equations, respectively. Lower mean creatinine excretion in these individuals may explain, in part, suboptimal agreement between estimated versus measured GFR. Inclusion of terms for ethnic and racial groups other than white and black might improve the performance of GFR estimating equations. PMID- 15800119 TI - Cellular response to injury in membranous nephropathy. AB - The pathogenesis of membranous nephropathy (MN) involves in situ formation of subepithelial immune deposits that produce glomerular injury by damaging and/or activating podocytes through complement-dependent processes. C5b-9 formation and insertion into podocyte cell membranes causes glomerular injury in MN. C5b-9 in sublytic quantities stimulates podocytes to produce proteases, oxidants, prostanoids, extracellular matrix components, and cytokines including TGF-beta. C5b-9 also causes alterations of the cytoskeleton that lead to abnormal distribution of slit diaphragm protein and detachment of viable podocytes that are shed into Bowman's space. These events result in disruption of the functional integrity of the glomerular basement membrane and the protein filtration barrier of podocytes with subsequent development of massive proteinuria. Complement components in proteinuric urine also induce tubular epithelial cell injury and mediate progressive interstitial disease in MN. Measurements of urinary C5b-9 or podocyte excretion in the urine may be useful in the diagnosis of MN and as measures of disease activity and response to therapy. Recent studies of cell cycle proteins and DNA damage in podocytes have clarified why podocytes fail to proliferate in response to C5b-9-mediated injury and podocyte loss in MN, resulting in the development of glomerular sclerosis and renal failure. Improved understanding of the role of complement in the pathogenesis of MN and of the cellular response to C5b-9 attack creates several new opportunities for therapeutic intervention that may benefit patients with MN in the future. PMID- 15800120 TI - Molecular pathomechanisms of membranous nephropathy: from Heymann nephritis to alloimmunization. AB - Membranous nephropathy (MN), the most common cause of idiopathic nephrotic syndrome in white adults, is characterized by an accumulation of immune deposits on the outer aspect of the glomerular basement membrane. In Heymann nephritis, the rat experimental model for MN, megalin--the target antigen of the nephritogenic antibodies--is expressed on the surface of podocytes, where immune complexes are formed, leading to complement activation and nephrotic-range proteinuria. However, megalin cannot be held responsible for human MN because it has not been found in human podocytes or detected in subepithelial immune deposits in patients with MN. Several potential antigens have been identified in so-called secondary forms of MN, but there is no real proof that these antigens are pathogenic. In a subgroup of infants with antenatal MN, neutral endopeptidase (NEP) has been identified as the first protein target on human podocytes of nephritogenic antibodies. The infants' mothers became immunized during pregnancy against NEP expressed on syncytiotrophoblastic cells because they were NEP deficient as a result of truncating mutations in the MME gene. Severity of neonatal renal disease was determined by the mothers' IgG response that led to the formation of the membrane attack complex of complement in the subepithelial deposits. Alloimmunization against NEP is a novel pathomechanism of MN that might also account for some cases of MN after renal or bone marrow transplantation. Other types of alloimmunization should be investigated in MN but also in other renal and nonrenal diseases, particularly those that affect the pediatric age. PMID- 15800121 TI - Race and electronically measured adherence to immunosuppressive medications after deceased donor renal transplantation. AB - Nonadherence to immunosuppressive medications may partly explain the worse allograft outcomes among black recipients of renal transplants. In a prospective cohort study of recipients of deceased donor renal transplants, microelectronic cap monitors were placed on bottles of one immunosuppressive medication to (1) measure average daily percentage adherence during the first posttransplantation year and (2) determine the factors associated with adherence. A total of 278 transplant recipients who provided sufficient microelectronic adherence data were grouped into four categories of average daily percentage adherence: 95 to 100% adherence (41.0% of patients), 80 to 95% adherence (32.4%), 50 to 80% adherence (12.9%), and 0 to 50% adherence (13.7%). In the unadjusted ordinal logistic regression model, black race was associated with decreased adherence (odds ratio [OR], 0.43; 95% confidence interval [CI], 0.26 to 0.72; P = 0.001). Cause of renal disease, Powerful Others health locus of control, transplant center, and dosing frequency were also associated with adherence. After adjustment for transplant center and dosing frequency, the association between black race and decreased adherence was substantially attenuated (OR, 0.65; 95% CI, 0.38 to 1.14, P = 0.13). Transplant center (P = 0.003) and increased dosing frequency (OR, 0.43; 95% CI, 0.22 to 0.86, for three or four times per day dosing; OR, 2.35; 95% CI, 1.01 to 5.45, for daily dosing; versus two times per day dosing; P = 0.003) remained independently associated with adherence. Other baseline demographic, socioeconomic, medical, surgical, and psychosocial characteristics were not associated with adherence. The transplant center and dosing frequencies of immunosuppressive medications are associated with adherence and explain a substantial proportion of the race-adherence relationship. PMID- 15800122 TI - Economic impact of case-mix adjusting the dialysis composite rate. AB - The Medicare program reimburses dialysis providers a flat rate for a bundle of services that comprise the basic dialysis treatment. This payment system is being modified to incorporate case-mix adjustment for age and body size, which have been shown to influence dialysis costs. This study simulated the economic impact of the recently issued Medicare rule on case-mix adjustment by estimating the variation in payments across patients, facilities, and broad classes of facilities. Case-mix adjustment results in considerable patient-level variation in payments (dollar 12.99 SD in case-mix adjusted payments). The variation across dialysis facilities is smaller but still economically significant (dollar 3.77 SD). However, there was little evidence that particular classes of facilities (e.g., ownership, chain membership, size) will be substantially advantaged or disadvantaged by case-mix adjustment. There do seem to be modest changes in the regional distribution of payments. PMID- 15800123 TI - High lipoprotein(a) levels and small apolipoprotein(a) size prospectively predict cardiovascular events in dialysis patients. AB - Lipoprotein(a) [Lp(a)] levels are increased in dialysis patients, suggesting that they may play a role in the elevated atherosclerotic cardiovascular disease (ASCVD) risk in this population. Few prospective studies of Lp(a) level, apolipoprotein(a) [apo(a)] size, and ASCVD have been performed in the dialysis population. An inception cohort of 833 incident dialysis patients were followed prospectively. Baseline Lp(a) was measured by apo(a) size-independent ELISA and apo(a) size by Western blot after SDS-agarose gel electrophoresis. A combined prospective nonfatal and fatal ASCVD end point included myocardial infarction, coronary revascularization, cerebrovascular accident, carotid endarterectomy, peripheral revascularization, gangrene, or limb amputation. Survival analyses were performed with adjustment for baseline demographics, comorbid conditions, ASCVD risk factors, albumin, lipids, and C-reactive protein. Median follow-up was 27.4 mo, with 297 ASCVD events, 130 non-ASCVD deaths, and seven losses to follow up over 1649 person-years. In multivariate Cox regression models, both high Lp(a) concentration (>/=53 nmol/L) and low molecular weight (LMW) apo(a) isoforms (123 nmol/L, the relative hazard (RH) of ASCVD was 1.73 (P < 0.0005), compared with high molecular weight apo(a) and Lp(a) level <123 nmol/L. No interactions by age, race, gender, diabetes, or ASCVD were present. Both LMW apo(a) size and high Lp(a) level predict ASCVD risk in dialysis patients, but the association of ASCVD with LMW isoforms is stronger than the association with high Lp(a) concentration. PMID- 15800124 TI - Relative glomerular hyperfiltration in primary aldosteronism. AB - Experimental and clinical data suggest that primary aldosteronism (PA) may be associated with cardiovascular hypertrophy and fibrosis, in part independent of the BP level. Whether PA may also result in specific deleterious effects on the kidneys was less studied. In 25 patients with tumoral PA, renal studies (urinary excretion of proteins, GFR, and effective renal plasma flow [ERPF], as clearances of technetium-labeled diethylene triaminopentaacetic acid and 131I-ortho iodohippurate, respectively) were performed both before and 6 mo after surgical cure. A control group consisting of patients with essential hypertension (EH) was studied before and after 6 mo of antihypertensive therapy. At baseline, PA and EH patients were similar with respect to demographic data, duration and level of hypertension, and GFR and ERPF. Urinary excretion of albumin and beta2 microglobulin were higher in PA than EH (88 +/- 26 versus 39 +/- 12 and 0.91 +/- 0.23 versus 0.26 +/- 0.19 mg/24 h, respectively; both P < 0.05). Adrenalectomy was followed by a decrease in arterial BP (by 28 +/- 3/13 +/- 2 mmHg), urinary excretion of albumin and beta2 microglobulin (by 48 +/- 19 and 0.53 +/- 0.21 mg/24 h, respectively), and GFR and ERPF (by 15 +/- 3 and 54 +/- 15 ml/min per 1.73 m(2), respectively). In EH, a similar decrease in pressure was associated with a decrease in albuminuria but no change in GFR or ERPF. In 17 of the 25 PA patients who received a 6-mo treatment of spironolactone, both GFR and ERPF decreased in parallel with BP, similar to what was observed after surgery. These data suggest that PA was associated with relative hyperfiltration, unmasked after suppression of aldosterone excess. PMID- 15800125 TI - Vacuolar H+-ATPase d2 subunit: molecular characterization, developmental regulation, and localization to specialized proton pumps in kidney and bone. AB - The ubiquitous multisubunit vacuolar-type proton pump (H+- or V-ATPase) is essential for acidification of diverse intracellular compartments. It is also present in specialized forms at the plasma membrane of intercalated cells in the distal nephron, where it is required for urine acidification, and in osteoclasts, playing an important role in bone resorption by acid secretion across the ruffled border membrane. It was reported previously that, in human, several of the renal pump's constituent subunits are encoded by genes that are different from those that are ubiquitously expressed. These paralogous proteins may be important in differential functions, targeting or regulation of H+-ATPases. They include the d subunit, where d1 is ubiquitous whereas d2 has a limited tissue expression. This article reports on an investigation of d2. It was first confirmed that in mouse, as in human, kidney and bone are two of the main sites of d2 mRNA expression. d2 mRNA and protein appear later during nephrogenesis than does the ubiquitously expressed E1 subunit. Mouse nephron-segment reverse transcription-PCR revealed detectable mRNA in all segments except thin limb of Henle's loop and distal convoluted tubule. However, with the use of a novel d2-specific antibody, high intensity d2 staining was observed only in intercalated cells of the collecting duct in fresh-frozen human kidney, where it co-localized with the a4 subunit in the characteristic plasma membrane-enhanced pattern. In human bone, d2 co localized with the a3 subunit in osteoclasts. This different subunit association in different tissues emphasizes the possibility of the H+-ATPase as a future therapeutic target. PMID- 15800126 TI - Identifying individuals with a reduced GFR using ambulatory laboratory database surveillance. AB - The use of outpatient laboratory databases to identify people with a low GFR may be part of an effective strategy to increase their use of treatments to prevent kidney failure. All renal function data from 17 independent outpatient laboratories in Eastern Ontario were combined to determine the proportion of adults with at least one serum creatinine measurement during a 1-yr period. The detection rates of low GFR were measured using different algorithms, and what proportion of identified low GFR was transient was considered. Canadian census data were used to calculate rates and proportions. Renal function testing was common. Of the 1,090,000 adult residents, 32% of the entire population and 63% of seniors had at least one serum creatinine measured during the study year. Sixteen percent of the population (49% of those with tests performed) had at least one GFR <80 ml/min per 1.73 m2, 5% (16%) had at least one GFR <60 ml/min per 1.73 m2, and 0.6% (1.7%) had at least one GFR <30 ml/min per 1.73 m2. Low GFR were usually not transient: 68% of individuals with subsequent testing at least 30 d later had a similar or worse GFR. Ambulatory laboratory database case finding, particularly in older patients, seems to be a promising method for easily identifying large segments of the population with persistent reductions in GFR. Whether such identification leads to improved health outcomes warrants further study. PMID- 15800127 TI - Race, gender, and incident dialysis patients' reported health status and quality of life. AB - Patient-reported physical and mental health status, along with disease-specific concerns ("quality of life") reported by patients, provide information about experience on dialysis that can help to improve patient care. Among prevalent hemodialysis (HD) patients, blacks have been shown to report higher health status and quality of life scores than whites, but whether similar race differences characterize incident patients who start regular dialysis is not known. Health status and quality of life, measured by the Kidney Disease Quality of Life-Short Form, reported by black men, black women, white men, and white women who initiated HD and peritoneal dialysis (PD) in the Dialysis Morbidity and Mortality Study (DMMS) Wave 2 were examined. After adjustment for patients' sociodemographic and clinical characteristics, the only race differences observed were that among HD patients, black women scored highest on the burden of kidney disease measure (lower perceived burden), and among PD patients, black men had the lowest satisfaction with care scores. It is possible that differences between blacks' and whites' reported health status and quality of life become more evident as patients' dialysis vintage increases, a hypothesis that requires following a well-characterized incident cohort over time. These findings add to existing evidence that minority patients on dialysis report less satisfaction with their care than do white patients, an important area for continued study. PMID- 15800128 TI - Profound mishandling of protein glycation degradation products in uremia and dialysis. AB - The aim of this study was to define the severe deficits of protein glycation adduct clearance in chronic renal failure and elimination in peritoneal dialysis (PD) and hemodialysis (HD) therapy using a liquid chromatography-triple quadrupole mass spectrometric detection method. Physiologic proteolysis of proteins damaged by glycation, oxidation, and nitration forms protein glycation, oxidation, and nitration free adducts that are released into plasma for urinary excretion. Inefficient elimination of these free adducts in uremia may lead to their accumulation. Patients with mild uremic chronic renal failure had plasma glycation free adduct concentrations increased up to five-fold associated with a decline in renal clearance. In patients with ESRD, plasma glycation free adducts were increased up to 18-fold on PD and up to 40-fold on HD. Glycation free adduct concentrations in peritoneal dialysate increased over 2- to 12-h dwell time, exceeding the plasma levels markedly. Plasma glycation free adducts equilibrated rapidly with dialysate of HD patients, with both plasma and dialysate concentrations decreasing during a 4-h dialysis session. It is concluded that there are severe deficits of protein glycation free adduct clearance in chronic renal failure and in ESRD on PD and HD therapy. PMID- 15800129 TI - Aminoglycosides induce acute cell signaling and chronic cell death in renal cells that express the calcium-sensing receptor. AB - The aminoglycoside antibiotics (AGAs) are calcium-sensing receptor (CaR) agonists that are toxic to the renal proximal tubule. Proximal tubule-derived opossum kidney (OK) cells express CaR-like proteins and respond to AGAs with intracellular Ca2+ mobilization and extracellular regulated protein kinase (ERK) phosphorylation. To examine the possible cellular basis of AGA toxicity, acute and chronic responses to AGA treatment in OK cells and in CaR stably transfected HEK-293 cells (CaR-HEK) were studied. Changes in cell-fate signaling, proliferation, and cell death were detected by semiquantitative Western blotting, Hoechst staining, cell counting, and FACS analysis. Confocal microscopy was used to study the relative internalization of fluorophore-labeled gentamicin in CaR transfected and -nontransfected cells. Here it is reported that the AGA neomycin and gentamicin elicit acute, phosphatidylinositol-3 kinase-dependent phosphorylation of Akt, glycogen synthase kinase 3beta, and p38 mitogen-activated protein kinase. After 24 h of gentamicin treatment, OK cell proliferation was observed, whereas after 4 d, the OK cells underwent cell death, an effect that was mimicked by the CaR agonists spermine and polyarginine. Furthermore, gentamicin elicited substantially more cell death in CaR-HEK cells than in nontransfected HEK-293 cells. The pan-specific caspase inhibitor Z-VAD significantly inhibited cell death in both OK and CaR-HEK cells. Finally, the intracellular uptake of Texas Red-labeled gentamicin was equivalent in both CaR transfected and vector-transfected HEK-293 cells, suggesting that the CaR does not enhance drug uptake. Together, these observations demonstrate that the AGAs induce both acute and chronic cell fate changes in OK cells and CaR-HEK cells and that the proximal tubular CaR is likely to contribute to signaling underlying the renal toxicity of the AGAs. PMID- 15800130 TI - Dialysate as food: combined amino acid and glucose dialysate improves protein anabolism in renal failure patients on automated peritoneal dialysis. AB - Protein-energy malnutrition as a result of anorexia frequently occurs in dialysis patients. In patients who are on peritoneal dialysis (PD), dialysate that contains amino acids (AA) improves protein anabolism when combined with a sufficient oral intake of calories. It was investigated whether protein anabolism can be obtained with a mixture of AA plus glucose (G) as a source of proteins and calories during nocturnal automated PD (APD). A random-order cross-over study was performed in eight APD patients to compare in two periods of 7 d each AA plus G dialysate obtained by cycler-assisted mixing of one bag of 2.5 L of AA (Nutrineal 1.1%, 27 g of AA) and four bags of 2.5 L of G (Physioneal 1.36 to 3.86%) versus G as control dialysate. Whole-body protein turnover was determined using a primed continuous infusion of L-[1-13C]leucine, and 24-h nitrogen balance studies were performed. During AA plus G dialysis, when compared with control, rates of protein synthesis were 1.20 +/- 0.4 and 1.10 +/- 0.2 micromol/kg per min leucine (mean +/- SD), respectively (NS), and protein breakdown rates were 1.60 +/- 0.5 and 1.72 +/- 0.3 micromol/kg per min (NS). Net protein balance (protein synthesis minus protein breakdown) increased on AA plus G in all patients (mean 0.21 +/- 0.12 micromol leucine/kg per min; P < 0.001). The 24-h nitrogen balance changed by 0.96 +/- 1.21 g/d, from -0.60 +/- 2.38 to 0.35 +/- 3.25 g/d (P = 0.061, NS), improving in six patients. In conclusion, APD with AA plus G dialysate improves protein kinetics. This dialysis procedure may improve the nutritional status in malnourished PD patients. PMID- 15800131 TI - Genetic coadaptation of the amylase gene system in Drosophila melanogaster: evidence for the selective advantage of the lowest AMY activity and of its epistatic genetic background. AB - In natural populations of Drosophila melanogaster, an amylase isozyme with the lowest alpha-amylase activity (AMY(1,1)) is predominant. To evaluate the selective significance of AMY(1,1) and its regulatory factor(s), we examined selection experiments in laboratory populations on two distinct food environments. After 300 generations, AMY(1,1) became predominant (89%) in a glucose (a product of AMY)-rich environment, while an isozyme with higher alpha amylase activity, AMY(1,6), became predominant (83%) in a starch (substrate)-rich environment. We found that the identical alleles of the amylase (Amy) gene, which encodes each of AMY(1,1) and AMY(1,6), were shared between the two populations in the different food environments, employing the nucleotide sequencing of the duplicated Amy genes. Nevertheless, AMY(1,6) homozygotes selected in the starch rich environment had a twofold higher AMY enzyme activity than those selected in the glucose-rich environment, suggesting a coadaptation of the coding region and its regulatory factor(s) on the genetic background. Such a difference in AMY enzyme activity was not detected between AMY(1,1) homozygotes, suggesting that the effect of the genetic background is epistatic. Our results indicate that natural selection is working on the Amy gene system as a whole for flies to adapt to the various food environments of local populations. PMID- 15800132 TI - Searching for methods to detect, prevent, and treat Alzheimer's disease. PMID- 15800133 TI - Mind, brain, and personality disorders. AB - OBJECTIVE: The use of the terms "mind" and "brain" in psychiatry is often associated with a set of polarities. Concepts such as environment, psychosocial, and psychotherapy are linked with "mind," while genes, biology, and medication are often associated with "brain." The author examines these dichotomies as they apply to personality disorders. METHOD: Research on antisocial and borderline personality disorders that is relevant to these dichotomies is evaluated. The implications of the findings for the understanding of pathogenesis and treatment are reconsidered. RESULTS: In the clinical setting, it is problematic to lump together terms such as "genes," "brain," and "biological" as though they are separate and distinct from terms such as "environment," "mind," and "psychosocial." These dichotomies are problematic, because genes and environment are inextricably intertwined in the pathogenesis of personality disorders, psychosocial experiences may result in permanent changes in the brain, and psychotherapy may have its effect by altering brain structure and function. The "theory of mind" is a useful construct for bridging "mind" and "brain" in the treatment of personality disorders. CONCLUSIONS: Severe personality disorders are best understood and treated without "either-or" dichotomies of brain and mind. Each domain has a different language, however, and the language of the mind is necessary to help the patient develop a theory of mind. PMID- 15800134 TI - The efficacy of light therapy in the treatment of mood disorders: a review and meta-analysis of the evidence. AB - OBJECTIVE: The purpose of this study was to assess the evidence base for the efficacy of light therapy in treating mood disorders. METHOD: The authors systematically searched PubMed (January 1975 to July 2003) to identify randomized, controlled trials of light therapy for mood disorders that fulfilled predefined criteria. These articles were abstracted, and data were synthesized by disease and intervention category. RESULTS: Only 13% of the studies met the inclusion criteria. Meta-analyses revealed that a significant reduction in depression symptom severity was associated with bright light treatment (eight studies, having an effect size of 0.84 and 95% confidence interval [CI] of 0.60 to 1.08) and dawn simulation in seasonal affective disorder (five studies; effect size=0.73, 95% CI=0.37 to 1.08) and with bright light treatment in nonseasonal depression (three studies; effect size=0.53, 95% CI=0.18 to 0.89). Bright light as an adjunct to antidepressant pharmacotherapy for nonseasonal depression was not effective (five studies; effect size=-0.01, 95% CI=-0.36 to 0.34). CONCLUSIONS: Many reports of the efficacy of light therapy are not based on rigorous study designs. This analysis of randomized, controlled trials suggests that bright light treatment and dawn simulation for seasonal affective disorder and bright light for nonseasonal depression are efficacious, with effect sizes equivalent to those in most antidepressant pharmacotherapy trials. Adopting standard approaches to light therapy's specific issues (e.g., defining parameters of active versus placebo conditions) and incorporating rigorous designs (e.g., adequate group sizes, randomized assignment) are necessary to evaluate light therapy for mood disorders. PMID- 15800135 TI - Hippocampus, IV: relational memory. PMID- 15800136 TI - Juliano Moreira (1873-1933): founder of scientific psychiatry in Brazil. PMID- 15800137 TI - Conversion from mild cognitive impairment to probable Alzheimer's disease predicted by brain magnetic resonance spectroscopy. AB - OBJECTIVE: Mild cognitive impairment has been regarded as a pre-Alzheimer condition, but some patients do not develop dementia. Given the available therapies for Alzheimer's disease, early diagnosis is of paramount importance. The authors' objective was to determine whether findings from magnetic resonance spectroscopy (MRS) of the hippocampus and other cortical areas would predict conversion from amnestic mild cognitive impairment to probable Alzheimer's disease. METHOD: A longitudinal inception cohort of 53 consecutive and incident subjects fulfilling the criteria of amnestic mild cognitive impairment was followed for a mean period of 3 years. At baseline, a neuropsychological examination (Mini-Mental State Examination, Blessed Dementia Rating Scale, Clinical Dementia Rating, verbal fluency test, and memory tests) and standard blood tests were performed, and three cortical areas were examined by proton MRS: left hippocampus, right parietal cortex, and left occipital cortex. The patients were evaluated periodically to detect conversion to probable Alzheimer's disease. The statistical analysis of predictions was based on receiver operating characteristic curves. RESULTS: By the follow-up assessment that occurred on average after 3 years, 29 patients (55%) had developed probable Alzheimer's disease. An occipital cortex N-acetylaspartate/creatine ratio < or =1.61 predicted dementia at 100% sensitivity and 75% specificity (area under the curve=0.91, 95% CI=0.80-0.97). The positive predictive value was 83%, and the negative predictive value was 100%, with an overall cross-validated classification accuracy of 88.7%. None of the values in the hippocampus and parietal cortex had significant predictive value. CONCLUSIONS: MRS of the brain performed on patients with mild cognitive impairment is a valuable tool in predicting conversion to probable Alzheimer's disease. Occipital values were more reliable than hippocampal values in this prediction. PMID- 15800138 TI - Does donepezil treatment slow the progression of hippocampal atrophy in patients with Alzheimer's disease? AB - OBJECTIVE: The only approved pharmacological approach for the symptomatic treatment of Alzheimer's disease in Japan is the use of a cholinesterase inhibitor, donepezil hydrochloride. Recent in vivo and in vitro studies raise the possibility that cholinesterase inhibitors can slow the progression of Alzheimer's disease. The purpose of the present study was to determine whether donepezil has a neuroprotective effect in Alzheimer's disease by using the rate of hippocampal atrophy as a surrogate marker of disease progression. METHOD: In a prospective cohort study, 54 patients with Alzheimer's disease who received donepezil treatment and 93 control patients with Alzheimer's disease who never received anti-Alzheimer drugs underwent magnetic resonance imaging (MRI) twice at a 1-year interval. The annual rate of hippocampal atrophy of each subject was determined by using an MRI-based volumetric technique. Background characteristics, age, sex, disease duration, education, MRI interval, apolipoprotein E (APOE) genotype, and baseline Alzheimer's Disease Assessment Scale score were comparable between the treated and control groups. RESULTS: The mean annual rate of hippocampal volume loss among the treated patients (mean=3.82%, SD=2.84%) was significantly smaller than that among the control patients (mean=5.04%, SD=2.54%). Upon analysis of covariance, where those confounding variables (age, sex, disease duration, education, MRI interval, APOE genotype, and baseline Alzheimer's Disease Assessment Scale score) were entered into the model as covariates, the effect of donepezil treatment on hippocampal atrophy remained significant. CONCLUSIONS: Donepezil treatment slows the progression of hippocampal atrophy, suggesting a neuroprotective effect of donepezil in Alzheimer's disease. PMID- 15800139 TI - Effect of raloxifene on prevention of dementia and cognitive impairment in older women: the Multiple Outcomes of Raloxifene Evaluation (MORE) randomized trial. AB - OBJECTIVE: This investigation examined whether raloxifene, a selective estrogen receptor modulator, affects the risk for Alzheimer's disease. METHOD: The Multiple Outcomes of Raloxifene Evaluation was a randomized, placebo-controlled trial among postmenopausal women with osteoporosis. The effect of raloxifene (60 or 120 mg/day) on vertebral fractures was the primary outcome. Development of mild cognitive impairment and dementia was a secondary outcome. Women were given clinical and cognitive evaluations at baseline and annually. After 3 years, among the 5,386 women enrolled at participating sites, those who had clinical symptoms of dementia or scored in the lowest 10th percentile on cognitive screening were evaluated by a blinded dementia specialist and had brain scans and laboratory tests to evaluate dementia etiology. Dementia was diagnosed by a blinded adjudication committee. RESULTS: Of the 5,386 women, 5,153 (95.7%) were classified as cognitively normal, 181 (3.4%) had mild cognitive impairment, and 52 (1.0%) had dementia, 36 with Alzheimer's disease. Compared to those taking placebo, women receiving 120 mg/day of raloxifene had a 33% lower risk of mild cognitive impairment (relative risk, 0.67; 95% confidence interval [CI], 0.46 0.98) and somewhat lower risks of Alzheimer's disease (relative risk=0.52, 95% CI=0.22-1.21) and any cognitive impairment (relative risk=0.73, 95% CI=0.53 1.01). Risks of mild cognitive impairment, Alzheimer's disease, and any impairment were not significantly different in the group taking 60 mg/day of raloxifene. CONCLUSIONS: Raloxifene at a dose of 120 mg/day, but not 60 mg/day, resulted in reduced risk of cognitive impairment in postmenopausal women. PMID- 15800140 TI - Neuropsychological differences between late-onset and recurrent geriatric major depression. AB - OBJECTIVE: Executive dysfunction, possibly related to vascular pathology, has been well documented in patients with a first episode of major depressive disorder in later life (late-onset geriatric major depression). However, it is unclear whether the neuropsychological presentation differs in patients with a lifetime history of major depressive disorder (recurrent geriatric major depressive disorder). The purpose of this study was to explore differences in neuropsychological function, symptoms, and cardiovascular comorbidity between patients with late-onset and recurrent geriatric major depression. METHOD: The study used a two-by-two factorial design in which one factor was current major depressive disorder (present versus absent) and the second factor was lifetime history of depression (present versus absent). Neuropsychological measures of executive functioning and episodic memory, as well as psychopathological symptoms and comorbid medical illness, were examined in a total of 116 older adults. RESULTS: Patients with late-onset major depressive disorder showed specific deficits in attention and executive function, whereas patients with recurrent major depressive disorder exhibited deficits in episodic memory. The rates of anhedonia and comorbid cardiovascular illness were higher in patients with late onset geriatric major depressive disorder. CONCLUSIONS: In contrast to recurrent geriatric major depressive disorder, late-onset major depressive disorder is characterized by specific deficits in tasks of attention and executive function, consistent with increased anhedonia and cardiovascular comorbidity. These findings, if confirmed, suggest that recurrent and late-onset geriatric major depressive disorder may represent distinct phenomenological entities. Such phenomenological differences as a function of lifetime history of major depression can guide research in the neurophysiology, prevention, and treatment of geriatric major depressive disorder. PMID- 15800141 TI - MRI hyperintensities and depressive symptoms in a community sample of individuals 60-64 years old. AB - OBJECTIVE: Previous studies have found associations of magnetic resonance imaging (MRI) signal hyperintensities with depression in the elderly. The present study investigates the association in a younger community sample (age 60-64 years) of depressed subjects and comparison groups for potential mediating and confounding variables. METHOD: A subsample of 475 persons 60-64 years of age from a larger community survey underwent brain MRI scans. White matter hyperintensities were quantified by using an automated procedure, and basal ganglia hyperintensities were quantified by using semiquantitative visual ratings. The study also assessed depressive symptoms and use of antidepressant medication. Potential mediating or confounding variables assessed included physical disability, hypertension, stroke, diabetes, head injury, cortisol, thyroid-stimulating hormone, cognitive functioning, smoking, and alcohol use. RESULTS: Depressive symptoms were found to be related to total brain white matter hyperintensities but not to basal ganglia hyperintensities. However, associations disappeared when statistical adjustment was made for physical disability and smoking. CONCLUSIONS: Depressive symptoms are related to white matter hyperintensities in mid-adult life in a community sample. Physical disability appears to play an important role in this association. PMID- 15800142 TI - Predictors of antidepressant use among older adults: have they changed over time? AB - OBJECTIVE: Antidepressant use increased substantially among older adults with the introduction of the new-generation medications such as the selective serotonin reuptake inhibitors. The authors analyzed data from two follow-up intervals-1986 1987 to 1989-1990 (interval 1) and 1992-1993 to 1996-1997 (interval 2)-from a community-based cohort of 4,162 older adults to determine predictors of future antidepressant use. METHOD: Information on antidepressant use, demographic and health characteristics, and categories of depressive symptoms-positive affect, negative affect, somatic complaints, and interpersonal problems-were obtained. Logistic regression was used to control simultaneously for multiple variables predicting antidepressant use during the two intervals. Repeated-measures logistic regression (with generalized estimating equations) was employed to model the probability of antidepressant use, with adjustment for the effect of time. RESULTS: Prior antidepressant use and white race were strong predictors of future use during both intervals. Negative affect was the only additional significant predictor of use during interval 1. In contrast, low positive affect scores, cognitive impairment, and poorer health were additional significant predictors during interval 2. In a repeated-measures model, race, prior antidepressant use, poor health, low positive affect scores, and somatic complaints varied as predictors over time. Negative affect and cognitive impairment were consistent predictors over time. CONCLUSIONS: The predictors of antidepressant use by older adults changed over time, with health-related measures of quality of life, such as positive affect, health status, and somatic complaints, becoming more prominent as predictors of use. PMID- 15800143 TI - Use of psychotherapy for depression in older adults. AB - OBJECTIVE: The authors examine national patterns in psychotherapy for older adults with a diagnosis of depression and analyze correlates of psychotherapy use that is consistent with Agency for Health Care Policy and Research guidelines for duration of treatment. METHOD: Linked Medicare claims and survey data from the 1992-1999 Medicare Current Beneficiary Survey were used. The data were merged with the Area Resource File to assess the effect of provider-supply influences on psychotherapy treatment. An episode-of-care framework approach was used to analyze psychotherapy use and treatment duration. Multiple logistic regression analysis was used to predict psychotherapy use and its consistency. RESULTS: The authors identified 2,025 episodes of depression treatment between 1992 and 1999. Overall, psychotherapy was used in 25% (N=474) of the episodes, with 68% of episodes with psychotherapy involving services received only from psychiatrists. (Percentages were weighted for the complex design of the Medicare Current Beneficiary Survey.) Use of psychotherapy was correlated with younger patient age, higher patient educational attainment, and availability of local psychotherapy providers. Among episodes in which psychotherapy was used, only a minority (33%, N=141) involved patients who remained in consistent treatment, defined as extending for at least two-thirds of the episode of depression. Availability of local providers was positively correlated with consistent psychotherapy use. In analyses with adjustment for provider-related factors, patients' socioeconomic and demographic characteristics did not affect the odds of receiving consistent psychotherapy. CONCLUSIONS: Use of psychotherapy remains uncommon among depressed older adults despite its widely acknowledged efficacy. Some of the disparities in psychotherapy utilization suggest supply-side barriers. Increasing the geographic availability of mental health care providers may be one way of increasing access to psychotherapy for depressed older adults. PMID- 15800145 TI - Critical periods of suicide risk in Huntington's disease. AB - Contrary to popular belief, receiving a diagnosis of a devastating fatal disease does not exacerbate, and may even alleviate, the risk of suicide. Suicidal ideation was examined in 4,171 individuals in the Huntington Study Group database. Participants were grouped according to a standardized neurological examination from 0 (i.e., normal examination) to 3 (definite Huntington's disease). Patients with an unequivocal diagnosis of Huntington's disease were further divided by stage of disease, from stage 1 (early) to stage 5 (end stage). Findings showed that the frequency of suicidal ideation doubled from 9.1% in at risk persons with a normal neurological examination to 19.8% in at-risk persons with soft neurological signs and increased to 23.5% in persons with "possible Huntington's disease." In persons with a diagnosis of Huntington's disease, 16.7% had suicidal ideation in stage 1, and 21.6% had suicidal ideation in stage 2, whereas the proportion of Huntington's disease patients with suicidal ideation diminished thereafter. Findings suggest two critical periods for increased risk of suicide in Huntington's disease. The first critical period is immediately before receiving a formal diagnosis of Huntington's disease, and the second is in stage 2 of the disease, when independence diminishes. Although the underlying mechanisms of suicidal ideation in Huntington's disease are poorly understood, it is critical for health care providers to be aware of periods during which patients may be at an increased risk. PMID- 15800144 TI - Remission in depressed geriatric primary care patients: a report from the PROSPECT study. AB - OBJECTIVE: This study compared time to first remission for elderly depressed patients in primary care for practices that implemented a care management model versus those providing usual care. In addition, it sought to identify risk factors for nonremission that could guide treatment planning and referral to care managers or specialists. METHOD: Prevention of Suicide in Primary Care Elderly: Collaborative Trial (PROSPECT) data were analyzed. Participants were older patients (> or =60 years) selected following screening of 9,072 randomly identified primary care patients. The present analysis examined patients with major depression and a 24-item Hamilton Depression Rating Scale score of 18 or greater who were followed for at least 4 months (N=215). Primary care practices were randomly assigned to offer the PROSPECT intervention or usual care. The intervention consisted of services of trained care managers, who offered algorithm-based recommendations to physicians and helped patients with treatment adherence over 18 months. RESULTS: First remission occurred earlier and was more common among patients receiving the intervention than among those receiving usual care. For all patients, limitations in physical and emotional functions predicted poor remission rate. Patients experiencing hopelessness were more likely to achieve remission if treated in intervention practices. Similarly, the intervention was more effective in patients with low baseline anxiety. CONCLUSIONS: Longitudinal assessment of depression, hopelessness, anxiety, and physical and emotional functional limitations in depressed older primary care patients is critical. Patients with prominent symptoms or impairment in these areas may be candidates for care management or mental health care, since they are at risk for remaining depressed and disabled. PMID- 15800146 TI - Symptom fluctuation in eating disorders: correlates of diagnostic crossover. AB - OBJECTIVE: The course of anorexia nervosa often includes the emergence of bulimic symptoms and a crossover to the full syndrome of bulimia nervosa. However, clinicians' ability to predict who will develop bulimia nervosa is limited. The converse phenomenon, crossover from bulimia nervosa to anorexia nervosa, has not been investigated as thoroughly. The authors identified factors that are associated with crossover from anorexia nervosa to bulimia nervosa and from bulimia nervosa to anorexia nervosa. METHOD: All participants were from the International Price Foundation Genetic Study. Two groups were studied. The first comprised 88 individuals with an initial diagnosis of anorexia nervosa, of whom 32 developed bulimia nervosa. The second included 350 individuals with bulimia nervosa, of whom 93 developed anorexia nervosa. Several variables, including DSM IV axis I and II disorders and personality traits, were evaluated as potential predictors of crossover. RESULTS: For the majority of affected individuals, crossover occurred by the fifth year of illness. A low level of self-directedness was associated with crossover in both directions. Other factors differed by diagnosis: high parental criticism was associated with crossover from anorexia nervosa to bulimia nervosa, whereas alcohol abuse/dependence and a low level of novelty seeking were associated with crossover from bulimia nervosa to anorexia nervosa. CONCLUSIONS: Low self-directedness may be associated with diagnostic instability in general, whereas other specific factors are related to the direction of diagnostic crossover. These results indicate that personality and family characteristics may influence the course of eating disorders and may be informative for planning interventions. PMID- 15800147 TI - Three psychotherapies for anorexia nervosa: a randomized, controlled trial. AB - OBJECTIVE: Few randomized, controlled trials have examined the efficacy of treatments for anorexia nervosa. Cognitive behavior therapy and interpersonal psychotherapy are effective in a related disorder, bulimia nervosa. There are theoretical and treatment indications for these therapies in anorexia nervosa. METHOD: Fifty-six women with anorexia nervosa diagnosed by using strict and lenient weight criteria were randomly assigned to three treatments. Two were specialized psychotherapies (cognitive behavior therapy and interpersonal psychotherapy), and one was a control treatment combining clinical management and supportive psychotherapy (nonspecific supportive clinical management). Therapy consisted of 20 sessions over a minimum of 20 weeks. RESULTS: For the total study group (intent-to-treat group), there were significant differences among therapies in the primary global outcome measure. Nonspecific supportive clinical management was superior to interpersonal psychotherapy, while cognitive behavior therapy was intermediate, neither worse than nonspecific supportive clinical management nor better than interpersonal psychotherapy. For the women completing therapy, nonspecific supportive clinical management was superior to the two specialized therapies. CONCLUSIONS: The finding that nonspecific supportive clinical management was superior to more specialized psychotherapies was opposite to the primary hypothesis and challenges assumptions about the effective ingredients of successful treatments for anorexia nervosa. PMID- 15800148 TI - The incidence of anorexia nervosa on Curacao. AB - OBJECTIVE: Although anorexia nervosa was once thought to occur only in affluent societies, cases have now been documented across the globe. To examine whether anorexia nervosa emerges in societies undergoing socioeconomic transition, the authors studied the incidence of anorexia nervosa on the Caribbean island of Curacao. METHOD: The authors contacted the full range of community health and service providers on Curacao, including dietitians, school counselors, and all 82 general practitioners. They also studied inpatient records for 84,420 admissions to Curacao General Hospital and two private hospitals in 1995-1998. Probable incident subjects were interviewed. RESULTS: The incidence rates in 1995-1998 per 100,000 person-years for anorexia nervosa on Curacao were 1.82 (95% confidence interval [CI]=0.74-2.89) for the total population and 17.48 (95% CI=4.13-30.43) for the high-risk group of 15-24-year-old females. No cases were found among the majority black population. For the Curacao mixed and white population, the incidence rate per 100,000 person-years for anorexia nervosa was 9.08 (95% CI=3.71-14.45). CONCLUSIONS: The overall incidence of anorexia nervosa on Curacao is much lower than in the affluent societies of the United States and Western Europe. Within Curacao, sociocultural factors appear to be associated with differential incidence rates of anorexia nervosa. The incidence of anorexia nervosa among the majority black population is nil, while the incidence among the minority mixed and white population on Curacao is similar to that of the United States and the Netherlands. PMID- 15800149 TI - Anorexia nervosa mortality in Northeast Scotland, 1965-1999. AB - OBJECTIVE: Most previous studies of mortality in anorexia nervosa patients have shown an increased risk of premature death but have been limited by methodological constraints. This study aimed to overcome some of these constraints by having a large original sample size, diagnosis confirmed by case note review, a long duration of follow-up, and a clear base population. METHOD: The authors identified 524 anorexia nervosa cases seen in specialist services in Northeast Scotland; anorexia nervosa diagnosis was confirmed by scrutinizing case notes. Those who had died were identified from the National Health Service register or register of deaths. The death rates and causes of death were analyzed. RESULTS: Twenty-three patients died, giving a crude death rate of 4.4% and a standardized mortality rate of 3.3 (95% CI=2.2-4.9). In only one-third of the cases was anorexia nervosa on the death certificate, but an eating disorder or other psychiatric pathology probably contributed to several of the other deaths. Older age at the time the patient was seen at the specialist service was the only identifiable risk factor in the group of patients who died. The median length of time between diagnosis and death was 11 years. CONCLUSIONS: Anorexia nervosa is associated with increased risk of premature death. It is possible that death rates could be reduced by early diagnosis and by long-term specialist care. PMID- 15800150 TI - Neuropsychological impairment and its neurological correlates in adult offspring with heightened risk for schizophrenia and affective psychosis. AB - OBJECTIVE: Schizophrenia is generally considered to be a neurodevelopmental disorder reflected in findings of neuropsychological impairments and neurological abnormality in patients and their relatives. The authors investigated whether neuropsychological impairments are related to neurological abnormality and whether such deficits also characterize risk for affective psychosis. METHOD: In a longitudinal study with a 93% rate of effective follow-up, the authors investigated neuropsychological impairment and its relation to neurological abnormality at a mean age of 22.3 years in 74 offspring of mothers with a history of psychotic disorders (38 offspring with heightened risk for schizophrenia and 36 with risk for affective psychosis) and 88 normal-risk offspring born to mothers with no history of psychosis. RESULTS: Offspring with genetically heightened risk for schizophrenia showed significantly impaired verbal memory, selective attention, and grammatical reasoning, compared with normal-risk offspring. Having impaired verbal memory, attention, and grammatical reasoning functions identified a significantly larger subgroup (16%) among offspring with heightened risk for schizophrenia than among offspring with heightened risk for affective psychosis (0%) and among normal-risk offspring (3%). Multiple neuropsychological functions were significantly related to neurological abnormality in offspring with heightened risk for schizophrenia and in normal risk offspring but not among offspring with heightened risk for affective psychosis. The extension of schizophrenia and affective psychosis risk groups to include additional offspring of mothers with psychosis-spectrum disorders yielded results similar to those for the core risk groups. CONCLUSIONS: The neurocognitive dysfunction attending heightened risk for schizophrenia is likely based on genetically mediated neurodevelopmental factors, with schizophrenia and affective psychosis belonging to different biological spheres. PMID- 15800151 TI - Maternal exposure to toxoplasmosis and risk of schizophrenia in adult offspring. AB - OBJECTIVE: The authors examined the relationship between maternal antibody to toxoplasmosis and the risk of schizophrenia and other schizophrenia spectrum disorders in offspring. Toxoplasmosis is known to adversely affect fetal brain development. METHOD: In a nested case-control design of a large birth cohort born between 1959 and 1967, the authors conducted serological assays for Toxoplasma antibody on maternal serum specimens from pregnancies giving rise to 63 cases of schizophrenia and other schizophrenia spectrum disorders and 123 matched comparison subjects. Toxoplasma immunoglobulin (Ig)G antibody was quantified by using the Sabin-Feldman dye test. The Ig titers were classified into three groups: negative (<1:16) (reference), moderate (1:16-1:64), and high (> or =1:128). RESULTS: The adjusted odds ratio of schizophrenia/schizophrenia spectrum disorders for subjects with high maternal Toxoplasma IgG antibody titers was 2.61 (95% confidence interval=1.00-6.82). There was no association between moderate Toxoplasma Ig antibody titers and the risk of schizophrenia/spectrum disorders. CONCLUSIONS: These findings suggest that maternal exposure to toxoplasmosis may be a risk factor for schizophrenia. The findings may be explained by reactivated infection or an effect of the antibody on the developing fetus. Given that toxoplasmosis is a preventable infection, the findings, if replicated, may have implications for reducing the incidence of schizophrenia. PMID- 15800152 TI - Psychosis in the community and in prisons: a report from the British National Survey of psychiatric morbidity. AB - OBJECTIVE: Reports of increased rates of psychosis in prisons could be due to sampling and ascertainment differences. The authors compared two samples of subjects 16-64 years of age: those from the general population of residents in Great Britain and prisoners in England and Wales. METHOD: A random sample of remanded and sentenced male and female prisoners (N=3,142) and a two-phase, cross sectional random sample of household residents (N=10,108) were assessed with structured questionnaires and the semistructured Schedules for Clinical Assessment in Neuropsychiatry. RESULTS: The weighted prevalence of probable functional psychosis in the past year was 4.5 per thousand (95% CI=3.1 to 5.8) in the household survey. In the prison survey, the weighted prevalence was over 10 times greater: 52 per thousand (95% CI=45 to 60). One in four prisoners with a psychotic disorder had psychotic symptoms attributed to toxic or withdrawal effects of psychoactive substances. The proportion of subjects with specific types of hallucinations or delusions did not differ between prison and household psychosis cases. CONCLUSIONS: This large study using standardized comparisons showed that the prevalence of psychosis in prisons is substantially higher than in the community and is deserving of greater attention to treatment and prevention. Apart from a minority of prisoners with symptoms attributable to psychoactive substances, the clinical symptom profile of psychosis is the same in both settings. Longitudinal research is needed to better understand these prevalence differences. PMID- 15800153 TI - Impact of referral source and study applicants' preference for randomly assigned service on research enrollment, service engagement, and evaluative outcomes. AB - OBJECTIVE: The inability to blind research participants to their experimental conditions is the Achilles' heel of mental health services research. When one experimental condition receives more disappointed participants, or more satisfied participants, research findings can be biased in spite of random assignment. The authors explored the potential for research participants' preference for one experimental program over another to compromise the generalizability and validity of randomized controlled service evaluations as well as cross-study comparisons. METHOD: Three Cox regression analyses measured the impact of applicants' service assignment preference on research project enrollment, engagement in assigned services, and a service-related outcome, competitive employment. RESULTS: A stated service preference, referral by an agency with a low level of continuity in outpatient care, and willingness to switch from current services were significant positive predictors of research enrollment. Match to service assignment preference was a significant positive predictor of service engagement, and mismatch to assignment preference was a significant negative predictor of both service engagement and employment outcome. CONCLUSIONS: Referral source type and service assignment preference should be routinely measured and statistically controlled for in all studies of mental health service effectiveness to provide a sound empirical base for evidence-based practice. PMID- 15800154 TI - Phenylthiocarbamide perception in patients with schizophrenia and first-degree family members. AB - OBJECTIVE: The inability to taste phenylthiocarbamide (PTC) has been associated with medical and neurological illnesses not typically related to taste. The authors examined PTC sensitivity in schizophrenia patients and their non-ill relatives to determine whether this represented a vulnerability marker. METHOD: PTC sensitivity was assessed in 42 schizophrenia patients, 23 healthy comparison subjects, and 12 first-degree relatives of the patients. RESULTS: More nontasters were found among patients and family members than healthy comparison subjects. Among patients, nontasters had more positive symptoms. Differences were not explained by sex, age, medication, smoking, or cognitive impairment. CONCLUSIONS: The prevalence of PTC nontasters was greater among schizophrenia patients and non ill first-degree family members. Phenotypic variation in PTC sensitivity is genetic in origin. This suggests a higher risk for illness among subjects with recessive alleles. PMID- 15800155 TI - Further evidence for altered cerebellar neuronal integrity in schizophrenia. AB - OBJECTIVE: The authors' goal was to investigate the distribution of metabolites and voxel composition in the pons and three cerebellar subregions and compare metabolite integral values and differences in voxel composition between patients with schizophrenia and healthy subjects. METHOD: Proton magnetic resonance spectroscopic imaging was used to study the cerebellum and pons of 14 patients with schizophrenia and 14 healthy comparison subjects. RESULTS: The voxel composition was not significantly different between the groups, but the patients with schizophrenia had significantly lower N-acetylaspartate levels in the cerebellar cortex and vermis. CONCLUSIONS: The lower integral value of N acetylaspartate in the cerebellar cortex and the vermis of patients with schizophrenia supports the theory of a dysfunctional corticocerebellar-thalamic cortical circuit in schizophrenia. PMID- 15800156 TI - The longitudinal course of thought disorder in geriatric patients with chronic schizophrenia. AB - OBJECTIVE: This study longitudinally assessed previously validated dimensions of thought disorder--verbal underproductivity and disconnection--in geriatric schizophrenia and replicated previous cross-sectional differences in communication disorders. METHOD: Two hundred twenty patients with chronic schizophrenia were assessed with the Thought, Language, and Communication Scale over a mean follow-up period of 2.3 years. RESULTS: Patients had increases in verbal underproductivity but stable scores on disconnection over the follow-up period. Worsening in verbal underproductivity was associated with concurrent worsening in Mini-Mental State Examination scores and older age at baseline. CONCLUSIONS: Verbal underproductivity worsens with aging and is associated with cognitive worsening, although disconnected speech is relatively stable. Some geriatric patients with schizophrenia may lose the skills necessary to report symptoms, leading to the impression that their clinical status is improving. PMID- 15800157 TI - Is combination olanzapine and antidepressant medication associated with a more rapid response trajectory than antidepressant alone? AB - OBJECTIVE: The authors' goal was to determine if prescription of antidepressant medication plus olanzapine initiates a more rapid response than prescription of antidepressant alone. METHOD: Twenty patients with major depression were studied. For 2 weeks the patients were blindly assigned to receive antidepressant plus olanzapine or antidepressant plus placebo. After 2 weeks, olanzapine augmentation was initiated for patients who did not improve with placebo augmentation. Response to medication was measured primarily by Hamilton Depression Rating Scale score. Other measures were the CORE, Clinical Global Impression, Beck Depression Inventory, and Daily Rating Schedule. RESULTS: Hamilton depression scores improved nonsignificantly in response to olanzapine combination therapy, but that trend was not evident on any secondary measure. Four patients who did not improve while receiving antidepressant and placebo showed rapid remission following late olanzapine augmentation. CONCLUSIONS: Failure to demonstrate any benefit from initial combination therapy may reflect an underpowered rather than a negative study. The distinct impact of late olanzapine augmentation suggests that pretreatment with an antidepressant may be required to facilitate a rapid antidepressant response to combined treatment. PMID- 15800158 TI - Suicide risk in placebo-controlled trials of treatment for acute manic episode and prevention of manic-depressive episode. AB - OBJECTIVE: The authors' goal was to investigate whether there is a greater suicide risk in the placebo arms of placebo-controlled studies of active medication for the treatment of acute manic episode and the prevention of manic/depressive episode. If so, this would be a strong ethical argument against the conduct of such studies. METHOD: All placebo-controlled, double-blind, randomized trials of medication for the treatment of acute manic episode and the prevention of manic/depressive episode that were part of a registration dossier submitted to the regulatory authority of the Netherlands, the Medicines Evaluation Board, between 1997 and 2003, were reviewed for occurrence of suicide and attempted suicide. RESULTS: In 11 placebo-controlled studies of the treatment of acute manic episode, including 1,506 patients (117 person-years) in the combined active compound group and 1,005 patients (71 person-years) in the combined placebo group, no suicides and no suicide attempts occurred. In four placebo-controlled studies of the prevention of manic/depressive episode, including 943 patients (406 person-years) in the combined active compound group and 418 patients (136 person-years) in the combined placebo group, two suicides (493/100,000 person-years of exposure) and eight suicide attempts (1,969/100,000 person-years of exposure) occurred in the combined active compound group, but no suicides and two suicide attempts (1,467/100,000 person-years of exposure) occurred in the combined placebo group. CONCLUSIONS: Concern about greater risk of suicide or attempted suicide in the placebo group should not be an argument against the conduct of placebo-controlled trials for these indications, provided that appropriate precautions are taken. PMID- 15800159 TI - Platelet serotonin reuptake inhibition and response to SSRIs in depressed adolescents. AB - OBJECTIVE: The authors examined platelet serotonin reuptake inhibition and response to selective serotonin reuptake inhibitor (SSRI) treatment in depressed adolescents. METHOD: Twenty-three depressed adolescents participating in pharmacokinetic studies of SSRIs had platelet serotonin reuptake measured before and after 14-28 days of treatment. The Clinical Global Impression (CGI) improvement rating was determined on the basis of all clinical information and was performed blind to the platelet data. RESULTS: Improvement in depressive symptoms as rated with the CGI improvement subscale was significantly associated with the percentage change in platelet serotonin reuptake inhibition from pre- to posttreatment. Improvement in depression was also associated with absolute decrease in platelet serotonin reuptake when adjusted for the magnitude of baseline reuptake. CONCLUSIONS: Platelet serotonin reuptake inhibition may be an appropriate surrogate biological marker for the pharmacodynamic activity of SSRIs in depressed adolescents. PMID- 15800160 TI - Effects of tryptophan depletion on serum levels of brain-derived neurotrophic factor in unmedicated patients with remitted depression and healthy subjects. AB - OBJECTIVE: Data suggest the involvement of serotonergic and neurotrophic systems in major depressive disorder. To investigate their potential interaction, the authors studied changes in serum levels of brain-derived neurotrophic factor (BDNF) during tryptophan depletion and sham depletion in unmedicated patients with remitted major depressive disorder and in a group of healthy comparison subjects. METHOD: Twenty-seven patients with remitted major depressive disorder and 20 healthy subjects underwent tryptophan depletion and sham depletion in a randomized, placebo-controlled, double-blind crossover study. Serum BDNF concentrations and plasma tryptophan concentrations as well as behavioral assessments were obtained. RESULTS: During tryptophan depletion, BDNF levels increased in healthy volunteers. By contrast, patients with remitted major depressive disorder were unable to mount this presumed compensatory response, and BDNF levels remained low in these patients. CONCLUSIONS: The results further substantiate the potential role of BDNF in major depressive disorder. PMID- 15800161 TI - Enhanced early morning salivary cortisol in neuroticism. AB - OBJECTIVE: Neuroticism is a predisposing factor for major depression. The increase in salivary cortisol that follows waking provides a reliable measure of adrenocortical activity, and this response is increased in recovered depressed patients. This study compared waking cortisol levels in healthy subjects with high and low levels of neuroticism without a previous history of depression. METHOD: Salivary cortisol levels were measured upon waking and at 15-minute intervals for the next hour in volunteers selected to have high (>19/23) or low (<4/23) neuroticism based on the Eysenck Personality Inventory. RESULTS: Subjects with high neuroticism showed significantly greater levels of salivary cortisol 30 minutes after waking, which were maintained for the next half hour. CONCLUSIONS: Abnormalities in waking cortisol are associated with neuroticism in a way similar to those seen in major depression. Elevated waking cortisol may represent a vulnerability marker for mood disorder. PMID- 15800162 TI - Multiple endocrine neoplasia type 1 presenting as psychosis. PMID- 15800163 TI - Neurotoxicity associated with free valproic acid. PMID- 15800164 TI - Increase in risperidone plasma level with lamotrigine. PMID- 15800165 TI - Liepmann's phenomenon during benzodiazepine withdrawal. PMID- 15800166 TI - Neuroleptic malignant syndrome induced by quetiapine and fluvoxamine. PMID- 15800167 TI - Mania in a boy treated with modafinil for narcolepsy. PMID- 15800168 TI - Elevated plasma ghrelin levels in night-eating syndrome. PMID- 15800169 TI - Pramipexole, ropinirole, and mania in Parkinson's disease. PMID- 15800170 TI - Clozapine-induced agranulocytosis after 11 years of treatment. PMID- 15800171 TI - 5-methoxy-N,N-diisopropyltryptamine-induced flashbacks. PMID- 15800172 TI - Do smokers who commit suicide have high blood levels of nicotine? PMID- 15800173 TI - Rate of tardive dyskinesia in hospitalized patients. PMID- 15800174 TI - Child psychopharmacology, effect sizes, and the big bang. PMID- 15800175 TI - Child psychopharmacology, effect sizes, and the big bang. PMID- 15800176 TI - Dementia with Lewy bodies, visual hallucinations, and medications. PMID- 15800177 TI - Child psychopharmacology, effect sizes, and the big bang. PMID- 15800178 TI - Intracytoplasmic sperm injection effects in infertile azh mutant mice. AB - Several reports in the literature describe men with infertility resulting from abnormal sperm head shape or decapitation defects of their spermatozoa. These defects are similar to those shown for the spermatozoa from azh (abnormal spermatozoon head shape) mice. The present study examines the efficiency and effects of intracytoplasmic sperm injection (ICSI) in successive generations of azh mice generated with this method. Three successive generations of azh mice were produced with ICSI. In all three ICSI series, more than 80% of 2-cell embryos were obtained, and more than 35% of embryos transferred gave rise to normal live offspring. In addition, ICSI was used to cross homozygous azh/azh males with homozygous azh/azh females, and live offspring were obtained. The ICSI derived males were tested for their fecundity and abnormalities of sperm morphology. Spermatozoa from ICSI-derived azh/+ males did not show any impairment of fecundity in in vitro fertilization. These spermatozoa successfully fertilized oocytes from both C57BL/6 and B6D2F1 females, with fertilization rates ranging from 70%- 92%. The proportion of morphologically normal spermatozoa was similar in azh/+ males from three successive generations of ICSI (57.8%, 54.8%, and 49.0%, respectively), and no differences were noted when comparing ICSI-derived males with males derived by mating (57.6%) and with wild-type controls (61.6%). Detailed analysis differentiating between specific types of anomalies of sperm morphology did not reveal significant differences among the examined groups. The results of the present study demonstrate that ICSI does not enhance the azh mutation phenotype in the offspring and brings no risks when applied continuously. Moreover, serial (successive generations) ICSI is highly efficient in maintaining valuable mice with fertility problems. PMID- 15800179 TI - Activated macrophages inhibit human cytotrophoblast invasiveness in vitro. AB - Pre-eclampsia is associated with inadequate cytotrophoblast invasion and remodeling of the uterine spiral arterioles, as well as by an aberrant maternal immune response. This study determined the effect of activated macrophages and one of its products, tumor necrosis factor (TNF)-alpha, on cytotrophoblast invasiveness. Coculture with human lipopolysaccharide-activated macrophages decreased the ability of immortalized HTR-8/ SVneo human trophoblast cells to invade through reconstituted extracellular matrix (P < 0.05). This effect of activated macrophages on trophoblast invasiveness was paralleled by abrogation of a 55-kDa caseinolytic activity corresponding to prourokinase plasminogen activator (pro-uPA) and an increased secretion of plasminogen activator inhibitor 1 (PAI1), as determined by gel zymography and ELISA, respectively. Coculture with nonactivated macrophages did not significantly affect trophoblast invasiveness or pro-uPA and PAI1 secretion. Activated macrophages secreted detectable levels of TNF, and administration of exogenous TNF significantly decreased trophoblast invasiveness (P < 0.05), increased the secretion of PAI1 (P < 0.01), and completely inhibited the pro-uPA-associated caseinolytic activity by binding to the TNF receptor 1. Moreover, addition of up to 10 ng/ml of TNF did not increase the rate of apoptosis in HTR-8/SVneo cells. Finally, the increased secretion of PAI1 by trophoblast cells cocultured with activated macrophages was significantly inhibited when a neutralizing anti-TNF antibody was added to the cocultures. These results suggest that the aberrant presence of activated macrophages around uterine vessels may contribute to inadequate trophoblast invasion and remodeling of the uterine spiral arterioles. Thus, the presence of activated macrophages may be important in the etiology of pre-eclampsia. PMID- 15800180 TI - Adherens junctions in myelinating Schwann cells stabilize Schmidt-Lanterman incisures via recruitment of p120 catenin to E-cadherin. AB - Schwann cell myelin contains highly compacted layers of membrane as well as noncompacted regions with a visible cytoplasm. One of these cytoplasmic compartments is the Schmidt-Lanterman incisure, which spirals through the compacted layers and is believed to help sustain the growth and function of compact myelin. Incisures contain adherens junctions (AJs), the key components of which are E-cadherin, its cytoplasmic partners called catenins, and F-actin. To explore in vivo the role of cadherin and catenins in incisures, E-cadherin mutant proteins that completely replace endogenous cadherin have been delivered to the cells using adenovirus. When the introduced cadherin lacked its extracellular domain, association of p120 catenin (p120ctn) with the cadherin did not occur, and incisures disappeared. Remarkably, the additional replacement of two phosphorylatable tyrosines by phenylalanine in the cytoplasmic tail of the mutant cadherin restored both p120ctn binding and incisure architecture, indicating that p120ctn recruitment is critical for incisures maintenance and might be regulated by phosphorylations. In addition, the ability of the p120ctn/cadherin complex to support incisures was blocked by mutation of the Rho GTPase regulatory region of the p120ctn, and downregulation of Rac1 activity at the junction reversed this inhibition. Because Rho GTPases regulate the state of the actin filaments, these findings suggest that one role of p120ctn in incisures is to organize the cytoskeleton at the AJ. Finally, developmental studies of Schwann cells demonstrated that p120ctn recruitment from the cytoplasm to the AJ occurs before the appearance of Rac1 GTPase and F-actin at the junction. PMID- 15800181 TI - Chronic antidepressants reduce depolarization-evoked glutamate release and protein interactions favoring formation of SNARE complex in hippocampus. AB - Glutamate neurotransmission was recently implicated in the action of stress and in antidepressant mechanisms. We report that chronic (not acute) treatment with three antidepressants with different primary mechanisms (fluoxetine, reboxetine, and desipramine) markedly reduced depolarization-evoked release of glutamate, stimulated by 15 or 25 mm KCl, but not release of GABA. Endogenous glutamate and GABA release was measured in superfused synaptosomes, freshly prepared from hippocampus of drug-treated rats. Interestingly, treatment with the three drugs only barely changed the release of glutamate (and of GABA) induced by ionomycin. In synaptic membranes of chronically treated rats we found a marked reduction in the protein-protein interaction between syntaxin 1 and Thr286-phosphorylated alphaCaM kinase II (alpha-calcium/calmodulin-dependent protein kinase II) (an interaction previously proposed to promote neurotransmitter release) and a marked increase in the interaction between syntaxin 1 and Munc-18 (an interaction proposed to reduce neurotransmitter release). Furthermore, we found a selective reduction in the expression level of the three proteins forming the core SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex. These findings suggest that antidepressants work by stabilizing glutamate neurotransmission in the hippocampus and that they may represent a useful tool for the study of relationship between functional and molecular processes in nerve terminals. PMID- 15800182 TI - A dissociation of encoding and retrieval processes in the human hippocampus. AB - The hippocampal formation performs two related but distinct memory functions: encoding of novel information and retrieval of episodes. Little evidence, however, resolves how these two processes are implemented within the same anatomical structure. Here we use high-resolution functional magnetic resonance imaging to show that distinct subregions of the hippocampus are differentially involved in encoding and retrieval. We found that regions early in the hippocampal circuit (dentate gyrus and CA fields 2 and 3) were selectively active during episodic memory formation, whereas a region later in the circuit (the subiculum) was active during the recollection of the learning episode. Different components of the hippocampal circuit likely contribute to different degrees to the two basic memory functions. PMID- 15800183 TI - High-frequency gamma-band activity in the basal temporal cortex during picture naming and lexical-decision tasks. AB - Gamma-band activity (GBA) in electroencephalograms (EEGs) has been shown to reflect various cognitive processes. GBA has typically been recorded in the 30-60 Hz range in scalp EEGs. Recently, task-related "high GBA" (HGBA) with frequencies up to 100 Hz has been observed in studies with invasive electrocorticograms (ECoGs). In the present study, we recorded ECoGs from the bilateral basal temporal cortices in a patient with epilepsy and evaluated the task-related HGBA (most prominently in the 80-120 Hz range) accompanying picture-naming and lexical decision tasks. We examined picture naming using two categories (line drawings of animals and tools). The lexical-decision task was performed using words and pseudowords of two distinct Japanese writing forms, kanji (morphograms) and kana (syllabograms). Task-related HGBA was observed bilaterally during the naming task. Recordings from some electrodes revealed significant differences in HGBA between animal and tool pictures. In contrast to the naming task, there was apparent left dominance in the lexical-decision task. Furthermore, significant differences in HGBA were observed between the Japanese kanji and kana words and between the kanji words and kanji pseudowords. A number of differences in the HGBA observed in the recordings from the basal temporal area were consistent with previous findings from neuroimaging and patient studies and suggest that HGBA is a good correlate of visual cognitive functions. PMID- 15800184 TI - Beta-adrenergic receptor activation facilitates induction of a protein synthesis dependent late phase of long-term potentiation. AB - Long-term potentiation (LTP) is activity-dependent enhancement of synaptic strength that can critically regulate long-term memory storage. Like memory, LTP exhibits at least two mechanistically distinct temporal phases. Early LTP (E-LTP) does not require protein synthesis, whereas the late phase of LTP (L-LTP), like long-term memory, requires protein synthesis. Hippocampal beta-adrenergic receptors can regulate expression of both E-LTP and long-term memory. Although beta-adrenergic receptor activation enhances the ability of subthreshold stimuli to induce E-LTP, it is unclear whether such activation can facilitate induction of L-LTP. Here, we use electrophysiological recording methods on mouse hippocampal slices to show that when synaptic stimulation that is subthreshold for inducing L-LTP is paired with beta-adrenergic receptor activation, the resulting LTP persists for over 6 h in area CA1. Like L-LTP induced by multiple trains of high-frequency electrical stimulation, this LTP requires protein synthesis. Unlike tetanus-induced L-LTP, however, L-LTP induced by beta adrenergic receptor activation during subthreshold stimulation appears to involve dendritic protein synthesis but not somatic transcription. Maintenance of this LTP also requires activation of extracellular signal-regulated kinases (ERKs). Thus, beta-adrenergic receptor activation elicits a type of L-LTP that requires translation and ERK activation but not transcription. This form of L-LTP may be a cellular mechanism for facilitation of behavioral long-term memory during periods of heightened emotional arousal that engage the noradrenergic modulatory system. PMID- 15800185 TI - Decisions under uncertainty: probabilistic context influences activation of prefrontal and parietal cortices. AB - Many decisions are made under uncertainty; that is, with limited information about their potential consequences. Previous neuroimaging studies of decision making have implicated regions of the medial frontal lobe in processes related to the resolution of uncertainty. However, a different set of regions in dorsal prefrontal and posterior parietal cortices has been reported to be critical for selection of actions to unexpected or unpredicted stimuli within a sequence. In the current study, we induced uncertainty using a novel task that required subjects to base their decisions on a binary sequence of eight stimuli so that uncertainty changed dynamically over time (from 20 to 50%), depending on which stimuli were presented. Activation within prefrontal, parietal, and insular cortices increased with increasing uncertainty. In contrast, within medial frontal regions, as well as motor and visual cortices, activation did not increase with increasing uncertainty. We conclude that the brain response to uncertainty depends on the demands of the experimental task. When uncertainty depends on learned associations between stimuli and responses, as in previous studies, it modulates activation in the medial frontal lobes. However, when uncertainty develops over short time scales as information is accumulated toward a decision, dorsal prefrontal and posterior parietal contributions are critical for its resolution. The distinction between neural mechanisms subserving different forms of uncertainty resolution provides an important constraint for neuroeconomic models of decision making. PMID- 15800186 TI - Amantadine inhibits NMDA receptors by accelerating channel closure during channel block. AB - The channel of NMDA receptors is blocked by a wide variety of drugs. NMDA receptor channel blockers include drugs of abuse that induce psychotic behavior, such as phencyclidine, and drugs with wide therapeutic utility, such as amantadine and memantine. We describe here the molecular mechanism of amantadine inhibition. In contrast to most other described channel-blocking molecules, amantadine causes the channel gate of NMDA receptors to close more quickly. Our results confirm that amantadine binding inhibits current flow through NMDA receptor channels but show that its main inhibitory action at pharmaceutically relevant concentrations results from stabilization of closed states of the channel. The surprising variation in the clinical utility of NMDA channel blockers may in part derive from their diverse effects on channel gating. PMID- 15800187 TI - Adaptivity of tuning functions in a generic recurrent network model of a cortical hypercolumn. AB - The representation of orientation information in the adult visual cortex is plastic as exemplified by phenomena such as perceptual learning or attention. Although these phenomena operate on different time scales and give rise to different changes in the response properties of neurons, both lead to an improvement in visual discrimination or detection tasks. If, however, optimal performance is indeed the goal, the question arises as to why the changes in neuronal response properties are so different. Here, we hypothesize that these differences arise naturally if optimal performance is achieved by means of different mechanisms. To evaluate this hypothesis, we set up a recurrent network model of a visual cortical hypercolumn and asked how each of four different parameter sets (strength of afferent and recurrent synapses, neuronal gains, and additive background inputs) must be changed to optimally improve the encoding accuracy of a particular set of visual stimuli. We find that the predicted changes in the population responses and the tuning functions were different for each set of parameters, hence were strongly dependent on the plasticity mechanism that was operative. An optimal change in the strength of the recurrent connections, for example, led to changes in the response properties that are similar to the changes observed in perceptual learning experiments. An optimal change in the neuronal gains led to changes mimicking neural effects of attention. Assuming the validity of the optimal encoding hypothesis, these model predictions can be used to disentangle the mechanisms of perceptual learning, attention, and other adaptation phenomena. PMID- 15800188 TI - Dissociable retrosplenial and hippocampal contributions to successful formation of survey representations. AB - During everyday navigation, humans encounter complex environments predominantly from a first-person perspective. Behavioral evidence suggests that these perceptual experiences can be used not only to acquire route knowledge but also to directly assemble map-like survey representations. Most studies of human navigation focus on the retrieval of previously learned environments, and the neural foundations of integrating sequential views into a coherent representation are not yet fully understood. We therefore used our recently introduced virtual reality paradigm, which provides accuracy and reaction-time measurements precisely indicating the emergence of survey knowledge, and functional magnetic resonance imaging while participants repeatedly encoded a complex environment from a first-person ground-level perspective. Before the experiment, we gave specific instructions to induce survey learning, which, based on the clear evidence for emerging survey knowledge in the behavioral data from 11 participants, proved successful. Neuroimaging data revealed increasing activation across sessions only in bilateral retrosplenial cortices, thus paralleling behavioral measures of map expertise. In contrast, hippocampal activation did not follow absolute performance but rather reflected the amount of knowledge acquired in a given session. In other words, hippocampal activation was most prominent during the initial learning phase and decayed after performance had approached ceiling level. We therefore conclude that, during navigational learning, retrosplenial areas mainly serve to integrate egocentric spatial information with cues about self-motion, whereas the hippocampus is needed to incorporate new information into an emerging memory representation. PMID- 15800189 TI - Molecular determinants for modulation of persistent sodium current by G-protein betagamma subunits. AB - Voltage-gated sodium channels are responsible for the upstroke of the action potential in most excitable cells, and their fast inactivation is essential for controlling electrical signaling. In addition, a noninactivating, persistent component of sodium current, I(NaP), has been implicated in integrative functions of neurons including threshold for firing, neuronal bursting, and signal integration. G-protein betagamma subunits increase I(NaP), but the sodium channel subtypes that conduct I(NaP) and the target site(s) on the sodium channel molecule required for modulation by Gbetagamma are poorly defined. Here, we show that I(NaP) conducted by Na(v)1.1 and Na(v)1.2 channels (Na(v)1.1 > Na(v)1.2) is modulated by Gbetagamma; Na(v)1.4 and Na(v)1.5 channels produce smaller I(NaP) that is not regulated by Gbetagamma. These qualitative differences in modulation by Gbetagamma are determined by the transmembrane body of the sodium channels rather than their cytoplasmic C-terminal domains, which have been implicated previously in modulation by Gbetagamma. However, the C-terminal domains determine the quantitative extent of modulation of Na(v)1.2 channels by Gbetagamma. Studies of chimeric and truncated Na(v)1.2 channels identify molecular determinants that affect modulation of I(NaP) located between amino acid residue 1890 and the C terminus at residue 2005. The last 28 amino acid residues of the C terminus are sufficient to support modulation by Gbetagamma when attached to the proximal C terminal domain. Our results further define the sodium channel subtypes that generate I(NaP) and identify crucial molecular determinants in the C-terminal domain required for modulation by Gbetagamma when attached to the transmembrane body of a responsive sodium channel. PMID- 15800190 TI - Retinoic acid-induced chromatin remodeling of mouse kappa opioid receptor gene. AB - The mouse kappa opioid receptor (KOR) gene is constitutively expressed in P19 embryonic stem cells but is first suppressed and reactivated during retinoic acid (RA)-induced neuronal differentiation. However, no RA response element (RARE) can be found in this gene regulatory region. The suppression and reactivation of the KOR gene in this neuronal differentiation model suggested chromatin remodeling occurred on this gene promoter triggered by RA induction. This study asks whether RA induces alteration in the nucleosomal structure of this gene promoter that has no apparent RARE and, if so, how RA remodels chromatin of this promoter. The results revealed two loose nucleosomes, N1 at -44 (3' boundary) from the transcription initiation site and N2 spanning the transcription initiation site, that are relevant to active transcription. RA formed a repressive chromatin configuration of this promoter by compacting nucleosome N1, followed by nucleosome N2 condensation. Chromatin immunoprecipitation assay demonstrated RA induced replacement of the c-Myc/Max complex with the Max/Mad1 complex on the E box located within nucleosome N1, coinciding with reduced Sp1 binding to GC boxes located within nucleosome N2 and recruitment of chromatin remodeling factor Brahma-related gene 1 (BRG-1) to this promoter. Consistently, histone deacetylation, Lys9 methylation, and hypophosphorylation of RNA polymerase II C terminal domain were detected on this promoter after RA treatment. It is concluded that RA induces KOR gene suppression, as early neuronal differentiation marker, by inducing substitution of c-Myc/Max with Max/Mad on the E box and by BRG-1 involved nucleosome recruitment and chromatin condensation, thereby abolishing Sp1 binding. PMID- 15800191 TI - Brain-derived neurotrophic factor enhances GABA release probability and nonuniform distribution of N- and P/Q-type channels on release sites of hippocampal inhibitory synapses. AB - Long-lasting exposures to brain-derived neurotrophic factor (BDNF) accelerate the functional maturation of GABAergic transmission in embryonic hippocampal neurons, but the molecular bases of this phenomenon are still debated. Evidence in favor of a postsynaptic site of action has been accumulated, but most of the data support a presynaptic site effect. A crucial issue is whether the enhancement of evoked IPSCs (eIPSCs) induced by BDNF is attributable to an increase in any of the elementary parameters controlling neurosecretion, namely the probability of release, the number of release sites, the readily releasable pool (RRP), and the quantal size. Here, using peak-scaled variance analysis of miniature IPSCs, multiple probability fluctuation analysis, and cumulative amplitude analysis of action potential-evoked postsynaptic currents, we show that BDNF increases release probability and vesicle replenishment with little or no effect on the quantal size, the number of release sites, the RRP, and the Ca2+ dependence of eIPSCs. BDNF treatment changes markedly the distribution of Ca2+ channels controlling neurotransmitter release. It enhances markedly the contribution of N- and P/Q-type channels, which summed to >100% ("supra-additivity"), and deletes the contribution of R-type channels. BDNF accelerates the switch of presynaptic Ca2+ channel distribution from "segregated" to "nonuniform" distribution. This maturation effect was accompanied by an uncovered increased control of N-type channels on paired-pulse depression, otherwise dominated by P/Q-type channels in untreated neurons. Nevertheless, BDNF preserved the fast recovery from depression associated with N-type channels. These novel presynaptic BDNF actions derive mostly from an enhanced overlapping and better colocalization of N- and P/Q-type channels to vesicle release sites. PMID- 15800192 TI - Step-response analysis of chemotaxis in Caenorhabditis elegans. AB - The sensorimotor transformation underlying Caenorhabditis elegans chemotaxis has been difficult to measure directly under normal assay conditions. Thus, key features of this transformation remain obscure, such as its time course and dependence on stimulus amplitude. Here, we present a comprehensive characterization of the transformation as obtained by inducing stepwise temporal changes in attractant concentration within the substrate as the worm crawls across it. We found that the step response is complex, with multiple phases and a nonlinear dependence on the sign and amplitude of the stimulus. Nevertheless, the step response could be reduced to a simple kinetic model that predicted the results of chemotaxis assays. Analysis of the model showed that chemotaxis results from the combined effects of approach and avoidance responses to concentration increases and decreases, respectively. Surprisingly, ablation of the ASE chemosensory neurons, known to be necessary for chemotaxis in chemical gradient assays, eliminated avoidance responses but left approach responses intact. These results indicate that the transformation can be dissected into components to which identified neurons can be assigned. PMID- 15800193 TI - A GIT1/PIX/Rac/PAK signaling module regulates spine morphogenesis and synapse formation through MLC. AB - Three of seven recently identified genes mutated in nonsyndromic mental retardation are involved in Rho family signaling. Two of the gene products, alpha p-21-activated kinase (PAK) interacting exchange factor (alphaPIX) and PAK3, form a complex with the synaptic adaptor protein G-protein-coupled receptor kinase interacting protein 1 (GIT1). Using an RNA interference approach, we show that GIT1 is critical for spine and synapse formation. We also show that Rac is locally activated in dendritic spines using fluorescence resonance energy transfer. This local activation of Rac is regulated by PIX, a Rac guanine nucleotide exchange factor. PAK1 and PAK3 serve as downstream effectors of Rac in regulating spine and synapse formation. Active PAK promotes the formation of spines and dendritic protrusions, which correlates with an increase in the number of excitatory synapses. These effects are dependent on the kinase activity of PAK, and PAK functions through phosphorylating myosin II regulatory light chain (MLC). Activated MLC causes an increase in dendritic spine and synapse formation, whereas inhibiting myosin ATPase activity results in decreased spine and synapse formation. Finally, both activated PAK and activated MLC can rescue the defects of GIT1 knockdown, suggesting that PAK and MLC are downstream of GIT1 in regulating spine and synapse formation. Our results point to a signaling complex, consisting of GIT1, PIX, Rac, and PAK, that plays an essential role in the regulation of dendritic spine and synapse formation and provides a potential mechanism by which alphaPIX and PAK3 mutations affect cognitive functions in mental retardation. PMID- 15800194 TI - Kv1.1 deletion augments the afferent hypoxic chemosensory pathway and respiration. AB - Mutations in the potassium channel gene Kv1.1 are associated with human episodic ataxia type 1 (EA-1) syndrome characterized by movement disorders and epilepsy. Ataxic episodes in EA-1 patients are often associated with exercise or emotional stress, which suggests a prominent role for the autonomic nervous system. Many of these alterations are reproduced in the Kv1.1-null mouse. Kv1.1 also regulates excitability of sensory neurons essential in cardiovascular and respiratory reflexes. We examined the neural control of the respiratory system of littermate wild-type (control) and Kv1.1-null mice during low O2 (hypoxia). Immunohistochemical studies demonstrated Kv1.1 in the afferent limb of the carotid body chemoreflex (the major regulator in the response to hypoxia), consisting of the carotid body, petrosal ganglion, and nucleus of the solitary tract (NTS). Respiration was examined by plethysmography. Null mice exhibited a greater increase in respiration during hypoxia compared with controls. In vitro carotid body sensory discharge during hypoxia was greater in null than control mice. In the caudal NTS, evoked EPSCs in brainstem slices were similar between control and null mice. However, the frequency of spontaneous and miniature EPSCs was greater in null mice. Null mice also exhibited more asynchronous release after a stimulus train. These results demonstrate the important role of Kv1.1 in afferent chemosensory activity and suggest that mutations in the human Kv1.1 gene have functional consequences during stress responses that involve respiratory reflexes. PMID- 15800196 TI - Neuronal mechanisms of repetition priming in occipitotemporal cortex: spatiotemporal evidence from functional magnetic resonance imaging and electroencephalography. AB - Repeated stimulus presentation (priming) is generally associated with a reduction in neuronal firing, macroscopically mirrored by a decrease in oscillatory electrophysiological markers as well as reduced hemodynamic responses. However, these repetition effects seem to be dependent on stimulus familiarity. We investigate the spatiotemporal correlates of repetition priming in cortical word recognition networks and their modulation by stimulus familiarity (i.e., words vs pseudowords). Event-related functional magnetic resonance imaging results show reduced activation for repeated words in occipitotemporal cortical regions. Electroencephalogram recordings reveal a significant reduction of induced gamma band responses (GBRs) between 200 and 350 ms after stimulus onset, accompanied by a decrease in phase synchrony between electrode positions. Pseudoword repetition, in contrast, leads to an activation increase in the same areas, to increased GBRs, and to an increased phase coupling. This spatiotemporal repetition by stimulus type interaction suggests that qualitatively distinct mechanisms are recruited during repetition priming of familiar and unfamiliar stimuli. Repetition of familiar stimuli leads to a "sharpening" of extrastriate object representations, whereas the repetition of unfamiliar stimuli results in the "formation" of a novel cortical network by means of synchronized oscillatory activity. In addition to isolating these mechanisms, the present study provides the first evidence for a possible link between induced electrophysiological and hemodynamic measures of brain activity. PMID- 15800195 TI - Relationship between membrane phosphatidylinositol-4,5-bisphosphate and receptor mediated inhibition of native neuronal M channels. AB - The relationship between receptor-induced membrane phosphatidylinositol-4'5' bisphosphate (PIP2) hydrolysis and M-current inhibition was assessed in single dissociated rat sympathetic neurons by simultaneous or parallel recording of membrane current and membrane-to-cytosol translocation of the fluorescent PIP2/inositol 1,4,5-trisphosphate (IP3)-binding peptide green fluorescent protein tagged pleckstrin homology domain of phospholipase C (GFP-PLCdelta-PH). The muscarinic receptor agonist oxotremorine-M produced parallel time- and concentration-dependent M-current inhibition and GFP-PLCdelta-PH translocation; bradykinin also produced parallel time-dependent inhibition and translocation. Phosphatidylinositol-4-phosphate-5-kinase (PI5-K) overexpression reduced both M current inhibition and GFP-PLCdelta-PH translocation by both oxotremorine-M and bradykinin. These effects were partly reversed by wortmannin, which inhibits phosphatidylinositol-4-kinase (PI4-K). PI5-K overexpression also reduced the inhibitory action of oxotremorine-M on PIP2-gated G-protein-gated inward rectifier (Kir3.1/3.2) channels; bradykinin did not inhibit these channels. Overexpression of neuronal calcium sensor-1 protein (NCS-1), which increases PI4 K activity, did not affect responses to oxotremorine-M but reduced both fluorescence translocation and M-current inhibition by bradykinin. Using an intracellular IP3 membrane fluorescence-displacement assay, initial mean concentrations of membrane [PIP2] were estimated at 261 microm (95% confidence limit; 192-381 microm), rising to 693 microm (417-1153 microm) in neurons overexpressing PI5-K. Changes in membrane [PIP2] during application of oxotremorine-M were calculated from fluorescence data. The results, taken in conjunction with previous data for KCNQ2/3 (Kv7.2/Kv7.3) channel gating by PIP2 (Zhang et al., 2003), accorded with the hypothesis that the inhibitory action of oxotremorine-M on M current resulted from depletion of PIP2. The effects of bradykinin require additional components of action, which might involve IP3 induced Ca2+ release and consequent M-channel inhibition (as proposed previously) and stimulation of PIP2 synthesis by Ca2+-dependent activation of NCS-1. PMID- 15800197 TI - Monosynaptic connections between pairs of spiny stellate cells in layer 4 and pyramidal cells in layer 5A indicate that lemniscal and paralemniscal afferent pathways converge in the infragranular somatosensory cortex. AB - Monosynaptic interlaminar connections between spiny stellate cells in layer 4 (L4), the main cortical recipient layer for thalamic projections, and pyramidal cells in layer 5A (L5A), one of the main cortical output layers, were examined anatomically and functionally by paired recordings in acute brain slices. The somata of pairs forming interlaminar L4-to-L5A connections were located predominantly close to or directly under the barrel-septum wall in layer 4. Superposition of spiny stellate axon arbors and L5A pyramidal cell dendritic arbors suggested an innervation domain underneath an L4 barrel wall. Functionally, the L4-to-L5A connections were of high reliability and relatively low efficacy, with a unitary EPSP amplitude of 0.6 mV, and the connectivity was moderately high (one in seven pairs tested was connected). The EPSP amplitude was weakly depressing (paired-pulse ratio of approximately 0.8) during repetitive presynaptic action potentials at 10 Hz. The existence of monosynaptic L4-to-L5A connections indicates that the specific "lemniscal" thalamic input from the ventro-basal nucleus of the thalamus to the cortex and the more unspecific "paralemniscal" afferent thalamic projections from the posterior medial nucleus of the thalamus merge already at an initial stage of cortical signal processing. These monosynaptic connections establish a monosynaptic coupling of the input to the cortex and its output, thereby effectively bypassing the supragranular layers. PMID- 15800198 TI - Sonic hedgehog has a dual effect on the growth of retinal ganglion axons depending on its concentration. AB - The stereotypical projection of retinal ganglion cell (RGC) axons to the optic disc has served as a good model system for studying axon guidance. By both in vitro and in vivo experiments, we show that a secreted molecule, Sonic hedgehog (Shh), may play a critical role in the process. It is expressed in a dynamic pattern in the ganglion cell layer with a relatively higher expression in the center of the retina. Through gel culture and stripe assays, we show that Shh has a dual effect on RGC axonal growth, acting as a positive factor at low concentrations and a negative factor at high concentrations. Results from time lapse video microscopic and stripe assay experiments further suggest that the effects of Shh on axons are not likely attributable to indirect transcriptional regulation by Shh. Overexpression of Shh protein or inhibition of Shh function inside the retina resulted in a complete loss of centrally directed projection of RGC axons, suggesting that precise regulation of Shh level inside the retina is critical for the projection of RGC axons to the optic disc. PMID- 15800199 TI - AMPA/kainate receptor-mediated downregulation of GABAergic synaptic transmission by calcineurin after seizures in the developing rat brain. AB - Hypoxia is the most common cause of perinatal seizures and can be refractory to conventional anticonvulsant drugs, suggesting an age-specific form of epileptogenesis. A model of hypoxia-induced seizures in immature rats reveals that seizures result in immediate activation of the phosphatase calcineurin (CaN) in area CA1 of hippocampus. After seizures, CA1 pyramidal neurons exhibit a downregulation of GABA(A) receptor (GABA(A)R)-mediated inhibition that was reversed by CaN inhibitors. CaN activation appears to be dependent on seizure induced activation of Ca2+-permeable AMPA receptors (AMPARs), because the upregulation of CaN activation and GABA(A)R inhibition were attenuated by GYKI 52466 [1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine hydrochloride] or Joro spider toxin. GABA(A)R beta2/3 subunit protein was dephosphorylated at 1 h after seizures, suggesting this subunit as a possible substrate of CaN in this model. Finally, in vivo administration of the CaN inhibitor FK-506 significantly suppressed hypoxic seizures, and posttreatment with NBQX (2,3-dihydroxy-6-nitro-7-sulfonyl-benzo[f]quinoxaline) or FK-506 blocked the hypoxic seizure-induced increase in CaN expression. These data suggest that Ca2+-permeable AMPARs and CaN regulate inhibitory synaptic transmission in a novel plasticity pathway that may play a role in epileptogenesis in the immature brain. PMID- 15800200 TI - Development of a new genetic model for absence epilepsy: spike-wave seizures in C3H/He and backcross mice. AB - To characterize the genetic basis of spike-wave discharges (SWDs) detected by electroencephalography (EEG) in C3H/He mice, substrains of C3H mice were evaluated by EEG and sensitivity to ethosuximide. Crosses with the SWD-negative strain C57BL/6J were performed to map the underlying gene(s). C3H/He substrains exhibited a modest incidence (average of 19 SWDs per hour) of 7-8 Hz SWDs when at rest, compared with the C3HeB/Fe subline (four SWDs per hour). In the mapping backcross, however, many mice showed a very high incidence (50-220 SWDs per hour) throughout the recording period. SWDs were first detected at 3.5 weeks of age, were associated with behavioral arrest, were suppressed by ethosuximide, and were strongest in the cerebral cortex and thalamus. The major C3H determinant of SWDs, spkw1 (spike-wave 1), mapped to chromosome (Chr 9), and together with a C57BL/6J determinant on Chr 8, spkw2, accounted for more than one-half of the phenotypic variation in the backcross mice. The modest SWD incidence in C3H/He mice and the high incidence in backcrosses implies that SWD could be a confounding variable for other behaviors. Because C3H/He mice have no other brain abnormalities, they are an attractive alternative for studying idiopathic absence epilepsy. PMID- 15800201 TI - Specializations for chromatic and temporal signals in human visual cortex. AB - Neurological case studies and qualitative measurements suggest that regions within human extrastriate cortex are specialized for different perceptual functions, including color. However, there are few quantitative measurements of human extrastriate color specializations. We studied the chromatic and temporal responses in several different clusters of human visual field maps using functional magnetic resonance imaging. Contrast response functions were measured for luminance [(L + M)-cone], red-green [(L - M)-cone] and blue-yellow (S-cone) modulations at various temporal frequencies. In primary visual cortex (V1), temporal responsivities to luminance and red-green modulations are approximately constant up to 10 Hz, but responsivities to blue-yellow modulations decrease significantly. In ventral occipital cortex (VO), all colors elicit strong responses, and, for each color, low temporal frequency modulations are more effective than high temporal frequency modulations. Hence, VO represents the full range of color information but does not respond well to rapid modulations. Conversely, in human motion-selective cortex (MT+) and V3A, blue-yellow modulations elicit very weak responses, whereas luminance and red-green high temporal frequency modulations are equally or more effective than low temporal frequency modulations. Hence, these dorsal occipital regions respond well to rapid modulations, but not all color information is represented. Similar to human motion perception, MT+ and V3A respond powerfully to all temporal frequencies but only to some colors. Similar to human color perception, VO responds powerfully to all colors but only to relatively low temporal frequencies. PMID- 15800202 TI - Neurodegenerative illness in transgenic mice expressing a transmembrane form of the prion protein. AB - Although PrP(Sc) is thought to be the infectious form of the prion protein, it may not be the form that is responsible for neuronal cell death in prion diseases. (Ctm)PrP is a transmembrane version of the prion protein that has been proposed to be a neurotoxic intermediate underlying prion-induced pathogenesis. To investigate this hypothesis, we have constructed transgenic mice that express L9R-3AV PrP, a mutant prion protein that is synthesized exclusively in the (Ctm)PrP form in transfected cells. These mice develop a fatal neurological illness characterized by ataxia and marked neuronal loss in the cerebellum and hippocampus. (Ctm)PrP in neurons cultured from transgenic mice is localized to the Golgi apparatus, rather than to the endoplasmic reticulum as in transfected cell lines. Surprisingly, development of the neurodegenerative phenotype is strongly dependent on coexpression of endogenous, wild-type PrP. Our results provide new insights into the cell biology of (Ctm)PrP, the mechanism by which it induces neurodegeneration, and possible cellular activities of PrP(C). PMID- 15800203 TI - Microanatomy of axon/glial signaling during Wallerian degeneration. AB - How do myelinated axons signal to the nuclei of cells that enwrap them? The cell bodies of oligodendrocytes and Schwann cells are segregated from axons by multiple layers of bimolecular lipid leaflet and myelin proteins. Conventional signal transduction strategies would seem inadequate to the challenge without special adaptations. Wallerian degeneration provides a model to study axon-to Schwann cell signaling in the context of nerve injury. We show a hitherto undetected rapid, but transient, activation of the receptor tyrosine kinase erbB2 in myelinating Schwann cells after sciatic nerve axotomy. Deconvolving microscopy using phosphorylation state-specific antibodies shows that erbB2 activation emanates from within the microvilli of Schwann cells, in direct contact with the axons they enwrap. To define the functional role of this transient activation, we used a small molecule antagonist of erbB2 activation (PKI166). The response of myelinating Schwann cells to axotomy is inhibited by PKI166 in vivo. Using neuron/Schwann cell cocultures prepared in compartmentalized cell culture chambers, we show that even transient activation of erbB2 is sufficient to initiate Schwann cell demyelination and that the initiating functions of erbB2 are localized to Schwann cells. PMID- 15800204 TI - Analysis of host response to bacterial infection using error model based gene expression microarray experiments. AB - A key step in the analysis of microarray data is the selection of genes that are differentially expressed. Ideally, such experiments should be properly replicated in order to infer both technical and biological variability, and the data should be subjected to rigorous hypothesis tests to identify the differentially expressed genes. However, in microarray experiments involving the analysis of very large numbers of biological samples, replication is not always practical. Therefore, there is a need for a method to select differentially expressed genes in a rational way from insufficiently replicated data. In this paper, we describe a simple method that uses bootstrapping to generate an error model from a replicated pilot study that can be used to identify differentially expressed genes in subsequent large-scale studies on the same platform, but in which there may be no replicated arrays. The method builds a stratified error model that includes array-to-array variability, feature-to-feature variability and the dependence of error on signal intensity. We apply this model to the characterization of the host response in a model of bacterial infection of human intestinal epithelial cells. We demonstrate the effectiveness of error model based microarray experiments and propose this as a general strategy for a microarray-based screening of large collections of biological samples. PMID- 15800205 TI - A GFP-based reporter system to monitor nonsense-mediated mRNA decay. AB - Aberrant mRNAs whose open reading frame (ORF) is truncated by the presence of a premature translation-termination codon (PTC) are recognized and degraded in eukaryotic cells by a process called nonsense-mediated mRNA decay (NMD). Here, we report the development of a reporter system that allows monitoring of NMD in mammalian cells by measuring the fluorescence of green fluorescent protein (GFP). The NMD reporter gene consists of a T-cell receptor-beta minigene construct, in which the GFP-ORF was inserted such that the stop codon of GFP is recognized as PTC. The reporter mRNA is therefore subjected to NMD, resulting in a low steady state mRNA level, an accordingly low protein level and hence a very low green fluorescence in normal, NMD-competent cells that express this reporter gene. We show that the inactivation of NMD by RNAi-mediated knockdown of the essential NMD factor hUpf1 or hSmg6 increases the NMD reporter mRNA level, resulting in a proportional increase of the green fluorescence that can be detected by flow cytometry, spectrofluorometry and fluorescence microscopy. With these properties, our GFP-based NMD reporter system could be used for large-scale screenings to identify NMD-inhibiting drugs or NMD-deficient mutant cells. PMID- 15800206 TI - Correcting errors in synthetic DNA through consensus shuffling. AB - Although efficient methods exist to assemble synthetic oligonucleotides into genes and genomes, these suffer from the presence of 1-3 random errors/kb of DNA. Here, we introduce a new method termed consensus shuffling and demonstrate its use to significantly reduce random errors in synthetic DNA. In this method, errors are revealed as mismatches by re-hybridization of the population. The DNA is fragmented, and mismatched fragments are removed upon binding to an immobilized mismatch binding protein (MutS). PCR assembly of the remaining fragments yields a new population of full-length sequences enriched for the consensus sequence of the input population. We show that two iterations of consensus shuffling improved a population of synthetic green fluorescent protein (GFPuv) clones from approximately 60 to >90% fluorescent, and decreased errors 3.5- to 4.3-fold to final values of approximately 1 error per 3500 bp. In addition, two iterations of consensus shuffling corrected a population of GFPuv clones where all members were non-functional, to a population where 82% of clones were fluorescent. Consensus shuffling should facilitate the rapid and accurate synthesis of long DNA sequences. PMID- 15800207 TI - Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces. AB - While it is universally accepted that intact RNA constitutes the best representation of the steady-state of transcription, there is no gold standard to define RNA quality prior to gene expression analysis. In this report, we evaluated the reliability of conventional methods for RNA quality assessment including UV spectroscopy and 28S:18S area ratios, and demonstrated their inconsistency. We then used two new freely available classifiers, the Degradometer and RIN systems, to produce user-independent RNA quality metrics, based on analysis of microcapillary electrophoresis traces. Both provided highly informative and valuable data and the results were found highly correlated, while the RIN system gave more reliable data. The relevance of the RNA quality metrics for assessment of gene expression differences was tested by Q-PCR, revealing a significant decline of the relative expression of genes in RNA samples of disparate quality, while samples of similar, even poor integrity were found highly comparable. We discuss the consequences of these observations to minimize artifactual detection of false positive and negative differential expression due to RNA integrity differences, and propose a scheme for the development of a standard operational procedure, with optional registration of RNA integrity metrics in public repositories of gene expression data. PMID- 15800208 TI - L1 retrotransposon-mediated stable gene silencing. AB - RNA interference (RNAi) is widely used for functional studies and has been proposed as a potential therapeutic agent. Current RNAi systems are largely efficient, but have limitations including transient effect, the need for viral handling and potential insertional mutations. Here, we describe a simple L1 retrotransposon-based system for the delivery of small interfering RNA (siRNA) and stable silencing in human cells. This system demonstrated long-term siRNA expression and significant reduction in both exogenous and endogenous gene expression by up to 90%. Further characterization indicated that retrotransposition occurred in a controlled manner such that essentially only one RNAi-cassette was integrated into the host genome and was sufficient for strong interference. Our system provides a novel strategy for stable gene silencing that is easy and efficient, and it may have potential applications for ex vivo and in vivo molecular therapy. PMID- 15800209 TI - Removal of mismatched bases from synthetic genes by enzymatic mismatch cleavage. AB - The success of long polynucleotide de novo synthesis is largely dependent on the quality and purity of the oligonucleotides used. Generally, the primary product of any synthesis reaction is directly cloned, and clones with correct products have to be identified. In this study, a novel strategy has been established for removing undesired sequence variants from primary gene synthesis products. Single base-pair mismatches, insertions and deletions were cleaved with specific endonucleases. Three different enzymes--T7 endonuclease I, T4 endonuclease VII and Escherichia coli endonuclease V--have been tested. As a model, a synthetic polynucleotide encoding the bacterial chloramphenicol-acetyltransferase (cat) was synthesized using different methods for one step polynucleotide synthesis based on ligation of oligonucleotides. The influence of enzymatic mismatch cleavage (EMC) as an error correction step on the frequency of correct products was analyzed by functional cloning of the synthetic cat and comparing the error rate with that of untreated products. Significant reduction of all mutation types was observed. Statistical analysis revealed that the T4 and E.coli endonucleases reduced the occurrence of mutations in cloned synthetic gene products. The EMC treatment was successful especially in the removal of deletions and insertions from the primary ligation products. PMID- 15800210 TI - Efficient and seamless DNA recombineering using a thymidylate synthase A selection system in Escherichia coli. AB - Lambda-red system-based recombinogenic engineering is a powerful new method to engineer DNA without the need for restriction enzymes or ligases. Here, we report the use of a single selectable marker to enhance the usefulness of this approach. The strategy is to utilize the thymidylate synthase A (thyA) gene, which encodes an enzyme involved in the synthesis of thymidine 5'-triphosphate, for both positive and negative selection. With this approach, we successfully created point mutations in plasmid and bacterial artificial chromosome (BAC) DNA containing the mouse Col10a1 gene. The results showed that the thyA selection system is highly efficient and accurate, giving an average of >90% selection efficiency. This selection system produces DNA that is free from permanent integration of unwanted sequences, thus allowing unlimited rounds of modifications if required. PMID- 15800211 TI - Dynamic relocalization of hOGG1 during the cell cycle is disrupted in cells harbouring the hOGG1-Cys326 polymorphic variant. AB - Numerous lines of evidence support the role of oxidative stress in different types of cancer. A major DNA lesion, 8-oxo-7,8-dihydroguanine (8-oxoG), is formed by reactive oxygen species in the genome under physiological conditions. 8-OxoG is strongly mutagenic, generating G.C-->T.A transversions, a frequent somatic mutation in cancers. hOGG1 was cloned as a gene encoding a DNA glycosylase that specifically recognizes and removes 8-oxoG from 8-oxoG:C base pairs and suppresses G.C-->T.A transversions. In this study, we investigated the subcellular localization and expression of hOGG1 during the cell cycle. Northern blots showed cell-cycle-dependent mRNA expression of the two major hOGG1 isoforms. By using a cell line constitutively expressing hOGG1 fused to enhanced green fluorescence protein (EGFP), we observed a dynamic relocalization of EGFP hOGG1 to the nucleoli during the S-phase of the cell cycle, and this localization was shown to be linked to transcription. A C/G change that results in an amino acid substitution from serine to cysteine in codon 326 has been reported as a genetic polymorphism and a risk allele for a variety of cancers. We investigated the cellular localization of the corresponding protein, hOGG1-Cys326, fused to EGFP and observed a dramatic effect on its localization that is explained by a change in the phosphorylation status of hOGG1. PMID- 15800212 TI - Torsional restraint: a new twist on frameshifting pseudoknots. AB - mRNA pseudoknots have a stimulatory function in programmed -1 ribosomal frameshifting (-1 PRF). Though we previously presented a model for how mRNA pseudoknots might activate the mechanism for -1 PRF, it did not address the question of the role that they may play in positioning the mRNA relative to the ribosome in this process [E. P. Plant, K. L. M. Jacobs, J. W. Harger, A. Meskauskas, J. L. Jacobs, J. L. Baxter, A. N. Petrov and J. D. Dinman (2003) RNA, 9, 168-174]. A separate 'torsional restraint' model suggests that mRNA pseudoknots act to increase the fraction of ribosomes directed to pause with the upstream heptameric slippery site positioned at the ribosome's A- and P-decoding sites [J. D. Dinman (1995) Yeast, 11, 1115-1127]. Here, experiments using a series of 'pseudo-pseudoknots' having different degrees of rotational freedom were used to test this model. The results of this study support the mechanistic hypothesis that -1 ribosomal frameshifting is enhanced by torsional resistance of the mRNA pseudoknot. PMID- 15800213 TI - A computational study of off-target effects of RNA interference. AB - RNA interference (RNAi) is an intracellular mechanism for post-transcriptional gene silencing that is frequently used to study gene function. RNAi is initiated by short interfering RNA (siRNA) of approximately 21 nt in length, either generated from the double-stranded RNA (dsRNA) by using the enzyme Dicer or introduced experimentally. Following association with an RNAi silencing complex, siRNA targets mRNA transcripts that have sequence identity for destruction. A phenotype resulting from this knockdown of expression may inform about the function of the targeted gene. However, 'off-target effects' compromise the specificity of RNAi if sequence identity between siRNA and random mRNA transcripts causes RNAi to knockdown expression of non-targeted genes. The complete off-target effects must be investigated systematically on each gene in a genome by adjusting a group of parameters, which is too expensive to conduct experimentally and motivates a study in silico. This computational study examined the potential for off-target effects of RNAi, employing the genome and transcriptome sequence data of Homo sapiens, Caenorhabditis elegans and Schizosaccharomyces pombe. The chance for RNAi off-target effects proved considerable, ranging from 5 to 80% for each of the organisms, when using as parameter the exact identity between any possible siRNA sequences (arbitrary length ranging from 17 to 28 nt) derived from a dsRNA (range 100-400 nt) representing the coding sequences of target genes and all other siRNAs within the genome. Remarkably, high-sequence specificity and low probability for off-target reactivity were optimally balanced for siRNA of 21 nt, the length observed mostly in vivo. The chance for off-target RNAi increased (although not always significantly) with greater length of the initial dsRNA sequence, inclusion into the analysis of available untranslated region sequences and allowing for mismatches between siRNA and target sequences. siRNA sequences from within 100 nt of the 5' termini of coding sequences had low chances for off-target reactivity. This may be owing to coding constraints for signal peptide-encoding regions of genes relative to regions that encode for mature proteins. Off-target distribution varied along the chromosomes of C.elegans, apparently owing to the use of more unique sequences in gene-dense regions. Finally, biological and thermodynamical descriptors of effective siRNA reduced the number of potential siRNAs compared with those identified by sequence identity alone, but off-target RNAi remained likely, with an off-target error rate of approximately 10%. These results also suggest a direction for future in vivo studies that could both help in calibrating true off-target rates in living organisms and also in contributing evidence toward the debate of whether siRNA efficacy is correlated with, or independent of, the target molecule. In summary, off-target effects present a real but not prohibitive concern that should be considered for RNAi experiments. PMID- 15800214 TI - Srs2 and RecQ homologs cooperate in mei-3-mediated homologous recombination repair of Neurospora crassa. AB - Homologous recombination and post-replication repair facilitate restart of stalled or collapsed replication forks. The SRS2 gene of Saccharomyces cerevisiae encodes a 3'-5' DNA helicase that functions both in homologous recombination repair and in post-replication repair. This study identifies and characterizes the SRS2 homolog in Neurospora crassa, which we call mus-50. A knockout mutant of N.crassa, mus-50, is sensitive to several DNA-damaging agents and genetic analyses indicate that it is epistatic with mei-3 (RAD51 homolog), mus-11 (RAD52 homolog), mus-48 (RAD55 homolog) and mus-49 (RAD57 homolog), suggesting a role for mus-50 in homologous recombination repair. However, epistasis evidence has presented that MUS50 does not participate in post-replication repair in N.crassa. Also, the N.crassa mus-25 (RAD54 homolog) mus-50 double mutant is viable, which is in contrast to the lethal phenotype of the equivalent rad54 srs2 mutant in S.cerevisiae. Tetrad analysis revealed that mus-50 in combination with mutations in two RecQ homologs, qde-3 and recQ2, is lethal, and this lethality is suppressed by mutation in mei-3, mus-11 or mus-25. Evidence is also presented for the two independent pathways for recovery from camptothecin-induced replication fork arrest: one pathway is dependent on QDE3 and MUS50 and the other pathway is dependent on MUS25 and RECQ2. PMID- 15800216 TI - Phenylthiocarbamide produces conditioned taste aversions in mice. AB - Previous work has demonstrated that SWR/J (SW) mice avoid phenylthiocarbamide (PTC) to a greater degree than C3HeB/FeJ mice in 48 h, two-bottle preference tests given in ascending series. The authors hypothesized, based also on previous work, that SW mice might form a conditioned taste aversion over time due to the toxic properties of PTC. We directly tested this hypothesis by attempting to condition a taste aversion to sucrose by injections of PTC. In experiment 1, PTC was nearly as effective as a strong dose of LiCl in reducing sucrose drinking. In experiment 2, the sucrose aversions were parametrically modified by both sucrose concentration and PTC dose, a hallmark of conditioned taste aversion. We conclude that PTC can cause a conditioned taste aversion and discuss the importance of considering toxic effects of aversive tastants when analyzing behavioral strain differences. PMID- 15800215 TI - The liver-enriched transcription factor CREB-H is a growth suppressor protein underexpressed in hepatocellular carcinoma. AB - We have previously characterized transcription factor LZIP to be a growth suppressor targeted by hepatitis C virus oncoprotein. In search of proteins closely related to LZIP, we have identified a liver-enriched transcription factor CREB-H. LZIP and CREB-H represent a new subfamily of bZIP factors. CREB-H activates transcription by binding to cAMP responsive element, box B, and ATF6 binding element. Interestingly, CREB-H has a putative transmembrane (TM) domain and it localizes ambiently to the endoplasmic reticulum. Proteolytic cleavage that removes the TM domain leads to nuclear translocation and activation of CREB H. CREB-H activates the promoter of hepatic gluconeogenic enzyme phosphoenolpyruvate carboxykinase. This activation can be further stimulated by cAMP and protein kinase A. CREB-H transcript is exclusively abundant in adult liver. In contrast, the expression of CREB-H mRNA is aberrantly reduced in hepatoma tissues and cells. The enforced expression of CREB-H suppresses the proliferation of cultured hepatoma cells. Taken together, our findings suggest that the liver-enriched bZIP transcription factor CREB-H is a growth suppressor that plays a role in hepatic physiology and pathology. PMID- 15800217 TI - Temporal changes in NCAM immunoreactivity during taste cell differentiation and cell lineage relationships in taste buds. AB - Neural cell adhesion molecule (NCAM) is a type III cell marker in the taste buds. In order to clarify the cell type of Mash1-expressing cells in taste buds, expression of NCAM was examined in Mash1-expressing taste cells of adult mice in comparison with gustducin- and T1r3-expressing cells, using a combination of NCAM immunohistochemistry and in situ hybridization. About 98% of Mash1-expressing cells were NCAM immunopositive (IP), suggesting that Mash1-expressing cells should be categorized as type III cells. Unexpectedly, small subsets of gustducin and T1r3-expressing cells were also found to be NCAM-IP, contradicting previous immunohistochemical studies in rats, in which gustducin-IP cells were observed specifically in type II cells, which do not have NCAM immunoreactivity. Examinations of developing taste buds showed temporal changes in the ratio of NCAM-IP cells in gustducin- and T1r3-expressing cells; the ratio of NCAM-IP cells in these gene-expressing cells were approximately 90% at 0.5 days after birth and decreased markedly during development. In contrast, the majority of Mash1 expressing cells showed constant NCAM immunoreactivity throughout development. In addition, BrdU-labeling experiments showed that the differentiation of Mash1 expressing cells precedes those of gustducin- and T1r3-expressing cells in taste buds of adult mice. These results suggest that T1r3- and gustducin-expressing cells are NCAM-IP at the beginning of cell differentiation, and that NCAM immunoreactivity in gustducin- and T1r3-expressing cells might remain from the previous developmental stage expressing Mash1. PMID- 15800218 TI - Evidence for multiple calcium response mechanisms in mammalian olfactory receptor neurons. AB - Olfactory receptor neurons employ a diversity of signaling mechanisms for transducing and encoding odorant information. The simultaneous activation of subsets of receptor neurons provides a complex pattern of activation in the olfactory bulb that allows for the rapid discrimination of odorant mixtures. While some transduction elements are conserved among many species, some species specificity occurs in certain features that may relate to their particular physiology and ecological niche. However, studies of olfactory transduction have been limited to a relatively small number of vertebrate and invertebrate species. To better understand the diversity and evolution of olfactory transduction mechanisms, we studied stimulus-elicited calcium fluxes in olfactory neurons from a previously unstudied mammalian species, the domestic cat. Isolated cells from cat olfactory epithelium were stimulated with odorant mixtures and biochemical agents, and cell responses were measured with calcium imaging techniques. Odorants elicited either increases or decreases in intracellular calcium; odorant induced calcium increases were mediated either by calcium fluxes through the cell membrane or by mobilization of intracellular stores. Individual cells could employ multiple signaling mechanisms to mediate responses to different odorants. The physiological features of these olfactory neurons suggest greater complexity than previously recognized in the role of peripheral neurons in encoding complex odor stimuli. The investigation of novel and unstudied species is important for understanding the mechanisms of odorant signaling that apply to the olfactory system in general and suggests both broadly conserved and species-specific evolutionary adaptations. PMID- 15800219 TI - Contribution of alpha-gustducin to taste-guided licking responses of mice. AB - We examined the necessity of alpha-gustducin, a G protein alpha-subunit expressed in taste cells, to taste-mediated licking responses of mice to sapid stimuli. To this end, we measured licking responses of alpha-gustducin knock-out (Gus-/-) mice and heterozygotic littermate controls (Gus+/-) to a variety of 'bitter', 'umami', 'sweet', 'salty' and 'sour' taste stimuli. All previous studies of how Gus-/- mice ingest taste stimuli have used long-term (i.e. 48 h) preference tests, which may be confounded by post-ingestive and/or experiential effects of the taste stimuli. We minimized these confounds by using a brief-access taste test, which quantifies immediate lick responses to extremely small volumes of sapid solutions. We found that deleting alpha-gustducin (i) dramatically reduced the aversiveness of a diverse range of 'bitter' taste stimuli; (ii) moderately decreased appetitive licking to low and intermediate concentrations of an 'umami' taste stimulus (monosodium glutamate in the presence of 100 microM amiloride), but virtually eliminated the normal aversion to high concentrations of the same taste stimulus; (iii) slightly decreased appetitive licking to 'sweet' taste stimuli; and (iv) modestly reduced the aversiveness of high, but not low or intermediate, concentrations of NaCl. There was no significant effect of deleting alpha-gustducin on licking responses to NH4Cl or HCl. PMID- 15800220 TI - Images in clinical medicine. Uremic frost. PMID- 15800221 TI - Raising the safety bar--the FDA's coxib meeting. PMID- 15800222 TI - Learning the value of drugs--is rofecoxib a regulatory success story? PMID- 15800223 TI - Veterans' mental health in the wake of war. PMID- 15800224 TI - Mental health care needs among recent war veterans. PMID- 15800225 TI - Standards of ethics at the National Institutes of Health. PMID- 15800226 TI - Comparison of warfarin and aspirin for symptomatic intracranial arterial stenosis. AB - BACKGROUND: Atherosclerotic intracranial arterial stenosis is an important cause of stroke. Warfarin is commonly used in preference to aspirin for this disorder, but these therapies have not been compared in a randomized trial. METHODS: We randomly assigned patients with transient ischemic attack or stroke caused by angiographically verified 50 to 99 percent stenosis of a major intracranial artery to receive warfarin (target international normalized ratio, 2.0 to 3.0) or aspirin (1300 mg per day) in a double-blind, multicenter clinical trial. The primary end point was ischemic stroke, brain hemorrhage, or death from vascular causes other than stroke. RESULTS: After 569 patients had undergone randomization, enrollment was stopped because of concerns about the safety of the patients who had been assigned to receive warfarin. During a mean follow-up period of 1.8 years, adverse events in the two groups included death (4.3 percent in the aspirin group vs. 9.7 percent in the warfarin group; hazard ratio for aspirin relative to warfarin, 0.46; 95 percent confidence interval, 0.23 to 0.90; P=0.02), major hemorrhage (3.2 percent vs. 8.3 percent, respectively; hazard ratio, 0.39; 95 percent confidence interval, 0.18 to 0.84; P=0.01), and myocardial infarction or sudden death (2.9 percent vs. 7.3 percent, respectively; hazard ratio, 0.40; 95 percent confidence interval, 0.18 to 0.91; P=0.02). The rate of death from vascular causes was 3.2 percent in the aspirin group and 5.9 percent in the warfarin group (P=0.16); the rate of death from nonvascular causes was 1.1 percent and 3.8 percent, respectively (P=0.05). The primary end point occurred in 22.1 percent of the patients in the aspirin group and 21.8 percent of those in the warfarin group (hazard ratio, 1.04; 95 percent confidence interval, 0.73 to 1.48; P=0.83). CONCLUSIONS: Warfarin was associated with significantly higher rates of adverse events and provided no benefit over aspirin in this trial. Aspirin should be used in preference to warfarin for patients with intracranial arterial stenosis. PMID- 15800227 TI - Sirolimus for Kaposi's sarcoma in renal-transplant recipients. AB - BACKGROUND: Recipients of organ transplants are susceptible to Kaposi's sarcoma as a result of treatment with immunosuppressive drugs. Sirolimus (rapamycin), an immunosuppressive drug, may also have antitumor effects. METHODS: We stopped cyclosporine therapy in 15 kidney-transplant recipients who had biopsy-proven Kaposi's sarcoma and began sirolimus therapy. All patients underwent an excisional biopsy of the lesion and one biopsy of normal skin at the time of diagnosis. A second biopsy was performed at the site of a previous Kaposi's sarcoma lesion six months after sirolimus therapy was begun. We examined biopsy specimens for vascular endothelial growth factor (VEGF), Flk-1/KDR protein, and phosphorylated Akt and p70S6 kinase, two enzymes in the signaling pathway targeted by sirolimus. RESULTS: Three months after sirolimus therapy was begun, all cutaneous Kaposi's sarcoma lesions had disappeared in all patients. Remission was confirmed histologically in all patients six months after sirolimus therapy was begun. There were no acute episodes of rejection or changes in kidney-graft function. Levels of Flk-1/KDR and phosphorylated Akt and p70S6 kinase were increased in Kaposi's sarcoma cells. The expression of VEGF was increased in Kaposi's sarcoma cells and even more so in normal skin cells around the Kaposi's sarcoma lesions. CONCLUSIONS: Sirolimus inhibits the progression of dermal Kaposi's sarcoma in kidney-transplant recipients while providing effective immunosuppression. PMID- 15800228 TI - Morphine, gabapentin, or their combination for neuropathic pain. AB - BACKGROUND: The available drugs to treat neuropathic pain have incomplete efficacy and dose-limiting adverse effects. We compared the efficacy of a combination of gabapentin and morphine with that of each as a single agent in patients with painful diabetic neuropathy or postherpetic neuralgia. METHODS: In this randomized, double-blind, active placebo-controlled, four-period crossover trial, patients received daily active placebo (lorazepam), sustained-release morphine, gabapentin, and a combination of gabapentin and morphine--each given orally for five weeks. The primary outcome measure was mean daily pain intensity in patients receiving a maximal tolerated dose; secondary outcomes included pain (rated according to the Short-Form McGill Pain Questionnaire), adverse effects, maximal tolerated doses, mood, and quality of life. RESULTS: Of 57 patients who underwent randomization (35 with diabetic neuropathy and 22 with postherpetic neuralgia), 41 completed the trial. Mean daily pain (on a scale from 0 to 10, with higher numbers indicating more severe pain) at a maximal tolerated dose of the study drug was as follows: 5.72 at baseline, 4.49 with placebo, 4.15 with gabapentin, 3.70 with morphine, and 3.06 with the gabapentin-morphine combination (P<0.05 for the combination vs. placebo, gabapentin, and morphine). Total scores on the Short-Form McGill Pain Questionnaire (on a scale from 0 to 45, with higher numbers indicating more severe pain) at a maximal tolerated dose were 14.4 with placebo, 10.7 with gabapentin, 10.7 with morphine, and 7.5 with the gabapentin morphine combination (P<0.05 for the combination vs. placebo, gabapentin, and morphine). The maximal tolerated doses of morphine and gabapentin were lower (P<0.05) with the combination than for each drug as single agent. At the maximal tolerated dose, the gabapentin-morphine combination resulted in a higher frequency of constipation than gabapentin alone (P<0.05) and a higher frequency of dry mouth than morphine alone (P<0.05). CONCLUSIONS: Gabapentin and morphine combined achieved better analgesia at lower doses of each drug than either as a single agent, with constipation, sedation, and dry mouth as the most frequent adverse effects. PMID- 15800229 TI - Blast injuries. PMID- 15800230 TI - Chronic inflammatory demyelinating polyneuropathy. PMID- 15800231 TI - Images in clinical medicine. Benign subcutaneous emphysema of the upper extremity. PMID- 15800232 TI - Case records of the Massachusetts General Hospital. Case 10-2005. A 73-year-old man with weakness and pain in the legs. PMID- 15800233 TI - Warfarin, aspirin, and intracranial vascular disease. PMID- 15800234 TI - Immunosuppressive drugs and the risk of cancer after organ transplantation. PMID- 15800235 TI - Combination therapy for neuropathic pain--which drugs, which combination, which patients? PMID- 15800236 TI - Amyotrophic lateral sclerosis--a new role for old drugs. PMID- 15800237 TI - Bacteremia among Kenyan Children. PMID- 15800238 TI - Adiposity and physical activity as predictors of mortality. PMID- 15800239 TI - Fecal DNA for colorectal-cancer screening. PMID- 15800240 TI - Levodopa and the progression of Parkinson's disease. PMID- 15800241 TI - Illness and Secrecy on the Supreme Court. PMID- 15800242 TI - Rivastigmine for dementia associated with Parkinson's disease. PMID- 15800243 TI - Medical education in Cuba. PMID- 15800244 TI - A Holter hazard. PMID- 15800245 TI - Stroke in the very elderly. PMID- 15800246 TI - Postcarotid endarterectomy hyperfusion or reperfusion syndrome. PMID- 15800247 TI - Embolus detection and differentiation using multifrequency transcranial Doppler. PMID- 15800248 TI - Emerging therapies: clopidogrel and aspirin. PMID- 15800250 TI - Editorial comment--transient ischemic attacks are emergencies. PMID- 15800251 TI - Editorial comment--time to burn the TOAST. PMID- 15800252 TI - Guidelines for the early management of patients with ischemic stroke: 2005 guidelines update a scientific statement from the Stroke Council of the American Heart Association/American Stroke Association. PMID- 15800253 TI - Clinical trial registration: a statement from the International Committee of Medical Journal Editors. PMID- 15800254 TI - Single-unit implants versus conventional treatments for compromised teeth: a brief review of the evidence. AB - The goal of preserving the natural dentition has long provided the foundation for clinical decision making in dentistry. Current trends in implant dentistry have weakened this paradigm as many practitioners have moved quickly to adopt implant dentistry as a new standard of care; however, the rapidity of this shift is a cause for concern among others. Many short-term studies have reported favorable data supporting the growth of single-unit implant dentistry, but the lack of standardized outcome evaluations and broadly conceived dimensions of performance makes it difficult to compare these reports. Thus, even with the exciting new treatment options implant dentistry offers patients and practitioners, all due consideration should first be given to treatments aimed at preserving and restoring compromised teeth before pursuing extraction and replacement. This article examines this premise from five perspectives: form, function, survival, management of complications, and quality of life. PMID- 15800255 TI - Understanding dental students' knowledge and perceptions of older people: toward a new model of geriatric dental education. AB - Increasing numbers of older people and the decreasing rates of edentulism highlight the importance of dental education that focuses on oral health and aging. This evaluation study assessed dental students' knowledge and beliefs about older people as well as their awareness of the biopsychosocial concerns that are potential barriers to oral health care. Dental students' (N=202) knowledge and perceptions of older people were evaluated before and after the first year of a new educational program. Students completed the Palmore Facts on Aging Quiz II (FAQ II) and answered questions about health problems and social concerns that may influence patient care. The intervention was twofold: 1) the CARES (Counseling, Advocacy, Referral, Education, and Service) Program, a clinical collaboration between the schools of Dental Medicine and Social Work, was initiated; and 2) all students were exposed to geriatric educational interventions. FAQ II scores did not significantly change, but dental students' awareness of mental health, independence, and social concerns increased between Times 1 and 2. The results of the study suggest that positive interactions with older adults by health care providers may depend more on positive perceptions toward older people than increased knowledge about aging. Future research will focus on positive experiences with older adults and attitudes of dental students toward the elderly. PMID- 15800256 TI - Comparing standardized measures of diligence and achievement with dental student academic performance. AB - Utilizing a reliable and valid instrument to measure a student's application of energy and effort towards a goal (diligence) or the ability to reach goals (achievement) would enable dental educators to anticipate academic performance. This knowledge could be used to better distribute faculty and educational resources, as additional tutors could be provided for students who score low on diligence and achievement instruments. In this study, fourth-year dental students completed the Diligence Inventory and the NachNaff Scale (which measures desire to achieve) immediately prior to graduation. The scores from both inventories were correlated with nine measures of academic performance. For males, the NachNaff Scale positively correlated (p<.05) only with the quantity of exceptional performance reports (EPR) and for females negatively correlated only with science DAT scores. The modest positive correlations, which differ for gender, suggest that the NachNaff may be of limited use to predict dental student performance. For males, the Total Diligence mean positively correlated with EPR and National Board Parts I and II scores. For females, the Total Diligence mean positively correlated with EPR and a predental biology-chemistry-physics grade composite. Given the simplicity of the Diligence Inventory and its significant correlations with academic performance demonstrated in this study, it appears to be a useful tool to gain insight into students' diligence in striving to obtain goals. PMID- 15800257 TI - Internal dental school environmental factors promoting faculty survival and success. AB - A career in dental academics offers ample rewards and challenges. To promote successful careers in dental education, prospective and new dental faculty should possess a realistic view of the dental school work environment, akin to the informed consent so valuable to patients and doctors. Self-assessment of personal strengths and weaknesses provides helpful information in matching faculty applicants with appropriate dental schools. Essential prehiring information also includes a written job description detailing duties and responsibilities, professional development opportunities, and job performance evaluation protocol. Prehiring awareness of what constitutes excellence in job performance will aid new faculty in allotting time to productive venues. New faculty should not rely solely on professional expertise to advance careers. Research and regular peer reviewed publications are necessary elements in academic career success, along with the ability to secure governmental, private foundation, and corporate grant support. Tactful self-promotion and self-definition to the dental school community are faculty responsibilities, along with substantial peer collaboration. The recruitment period is a singular opportunity to secure job benefits and privileges. It is also the time to gain knowledge of institutional culture and assess administrative and faculty willingness to collaborate on teaching, research, professional development, and attainment of change. Powerful people within dental schools and parent institutions may influence faculty careers and should be identified and carefully treated. The time may come to leave one's position for employment at a different dental school or to step down from full-time academics. Nonetheless, the world of dental and health professional education in 2005 is rapidly expanding and offers unlimited opportunities to dedicated, talented, and informed educators. PMID- 15800258 TI - Family medicine residency directors' knowledge and attitudes about pediatric oral health education for residents. AB - The Surgeon General's report on oral health called for improved education about oral health, a renewed understanding of relationships between oral and overall health, and an interdisciplinary approach to oral health involving primary care providers. This study examined the following: 1) family medicine residency directors' knowledge of preventive dental care, 2) status of an oral heath curriculum in family medicine residencies, and 3) the likelihood of initiating an oral health curriculum. We conducted a fifty-item survey of family medicine program directors emphasizing pediatric oral health assessed demographics, knowledge of preventive procedures, existing oral health curriculum, composition, and time commitment for an oral health curriculum. Directors returned 208 (45 percent) surveys. Approximately 95 percent agreed that oral health knowledge should be a component of residency training. Most programs are teaching anticipatory guidance. The mean time program directors felt should be given to an oral health curriculum was four hours. Program directors lack knowledge of preventive dental procedures and oral health care recommendations. Oral health care knowledge is felt to be an important component of residency training. Program directors need faculty development for a successful delivery of an oral health curriculum. PMID- 15800259 TI - The effects of finger rest positions on hand muscle load and pinch force in simulated dental hygiene work. AB - One of the techniques taught in dental and dental hygiene programs is to use finger rests to stabilize the instrument while performing dental scaling or other types of dental work. It is believed that finger rests may also reduce muscle stress and prevent injury due to muscle fatigue. In this study the effects of three different finger rest positions on hand muscle activity and thumb pinch force were compared. Twelve predental students performed simulated dental scaling tasks on a manikin using three different finger rest positions: 1) no finger rest, 2) one finger rest, and 3) two finger rests. Muscle activity and thumb pinch force were measured by surface electromyography and a pressure sensor, respectively. Using two finger rests was always associated with reduced thumb pinch force and muscle activity, as compared to not using any finger rests (p<0.05), while using one finger rest reduced thumb pinch force and muscle activity in most cases. Hence, using finger rests plays an important role in reducing the muscle load of the hand in students performing simulated dental hygiene work. It is concluded that dental and dental hygiene students may benefit from instructions for using finger rests at an early stage of their clinical training. Including biomechanical and ergonomic principles in dental and dental hygiene curricula will raise awareness of ergonomics among dental practitioners and help them incorporate these principles into daily practice. PMID- 15800260 TI - Dental education in India and Japan: implications for U.S. dental programs for foreign-trained dentists. AB - An understanding of international dental education systems is critical for the education of foreign-trained dentists in U.S. dental programs. However, there is little information on this topic. This article provides information regarding 1) dental history, 2) dental school system, 3) curriculum/examination at dental school, and 4) dental licensure in India and Japan. There are 185 dental schools in India and twenty-nine in Japan. The number of first-year dental students is 12,872 and 2,647 in India and Japan, respectively. A five-year dental education, which includes 4,035 curriculum hours, leads to the B.D.S. degree in India, whereas a six-year dental education program, which includes 5,060 curriculum hours, leads to the D.D.S. degree in Japan. No undergraduate predental study is needed prior to entry into dental school in both countries. In India, the entrance examination is extremely competitive; however, there is no nationwide licensure examination. In Japan, dental schools use more sophisticated dental materials and equipment in the clinical phases of the curriculum than in India, but there is no clinical examination at the time of graduation. Several implications for U.S. dental programs for foreign-trained dentists with respect to screening applicants and curriculum development are discussed. PMID- 15800261 TI - Use of the Internet among dental students in Turkey. AB - The extent to which dental students in Suleyman Demirel University (Turkey) use the Internet during their undergraduate dental training and their attitudes towards the Internet in general were assessed in this study. Ninety-three percent of the students use the Internet in various frequencies. Seven percent stated that they do not know how to use the Internet and, in this regard, the gender proportion was higher in the female students (p=0.026). In general, male students use the Internet more often than their female counterparts (p<0.001). Although a similar proportion of students (approximately 55 percent) use the Internet to retrieve information for general and dental topics, dental topics were searched in the Internet less frequently. To obtain information on dental topics, students generally referred to lecture handouts or textbooks (76 percent) while 32 percent accessed the Internet. Students who use the Internet to obtain information on dental topics most commonly preferred the university sites (71 percent), followed by search engines by performing key word searches (47 percent). The texts are the most commonly accessed materials (83 percent). Clinical photographs were accessed by 53 percent and histopathologic and radiographic materials by 14 percent of the students. PMID- 15800262 TI - Polygenic effects and cigarette smoking account for a portion of the familial aggregation of nuclear sclerosis. AB - Cataract is the most common cause of blindness worldwide. Nuclear cataract, an advanced stage of nuclear sclerosis, is the most common type of age-related cataract. The authors assessed data from 2,089 persons within 620 extended pedigrees who participated in the 1988-1990 Beaver Dam Eye Study in Wisconsin to determine whether the observed familial aggregation of nuclear sclerosis could be explained by inheritance of a major gene. Familial correlations were examined and segregation analyses were performed on nuclear sclerosis measurements adjusted for age, sex, and pack-years of cigarette smoking. There was modest correlation among close family members after adjustment for age, sex, and pack-years of cigarette smoking: 0.084 between parents and offspring, and 0.198 between sibling pairs. Although results do not support involvement of a single major locus in the etiology of nuclear sclerosis, models that allowed for familial correlation, attributable in part to polygenic effects, did provide a better fit to the observed data than models without a polygenic effect. This finding suggests that several genes of modest effect may influence development of nuclear lens opacity, possibly in conjunction with environmental factors. Cigarette smoking was an important covariate in these analyses. Overall, results highlight the complex etiology of nuclear sclerosis. PMID- 15800263 TI - Relative contributions of genes, environment, and interactions to blood lipid concentrations in a general adult population. AB - The authors evaluated the contributions of nine genetic (G) variants (selected from 275 single nucleotide polymorphisms in 11 reverse cholesterol transport pathway genes), five environmental (E) factors (selected from 10), and G x G, E x E, and G x E interactions in explaining population variance of blood lipid concentrations. Total cholesterol, triglycerides, and high density lipoprotein (HDL) cholesterol were measured, and low density lipoprotein (LDL) cholesterol and HDL cholesterol/LDL cholesterol ratio were calculated in a population-based random sample of 1,543 men and women in Geneva, Switzerland, aged 35-74 years in 1999-2001. Explained variances (R2) for HDL cholesterol/LDL cholesterol ratio, HDL cholesterol, and LDL cholesterol, respectively, were 34%, 33%, and 19%, decomposed into main effects of G (6%, 4%, and 5%) and E (25%, 28%, and 11%), with just 3%, 2%, and 3% due to G x G, E x E, and G x E interactions, respectively. Risk factor clustering was only moderate: 70% of study subjects had < or =3 variants, 75% had < or =2 environmental exposures, and 69% had < or =5 of both types of factors. Multiple genes with weak associations, together with more dominating environmental factors, are involved in determining blood lipid concentrations. Interactions added little explained variance. Increasing trends in hypercholesterolemia are attributable to environmental changes affecting populations as a whole. Reducing obesity and smoking and moderating alcohol intake in entire populations should remain the primary strategies for lipid control. PMID- 15800264 TI - The probability of giving birth among women who were born preterm or with impaired fetal growth: a Swedish population-based registry study. AB - The primary aim of this study was to investigate whether women born prematurely or with impaired fetal growth have a reduced probability of giving birth. Using Swedish population-based registries, the authors identified 148,281 women born in 1973-1975 for follow-up until 2001. Of these women, 4.1% were born preterm and 0.32% very preterm, 0.29% were born with a very low birth weight, and 5.4% were small for gestational age. Outcome measures were the hazard ratios for giving birth during the study period. Adjustments were made for socioeconomic factors. Very-low-birth-weight women displayed a reduced probability of giving birth (hazard ratio = 0.74, 95% confidence interval: 0.60, 0.91), most apparent among women aged 25 or more years. There were also tendencies of reduced hazard ratios of giving birth among women born preterm or very preterm in this age interval. Women born small for gestational age (below -2 standard deviations) seemed to be more likely to have given birth (hazard ratio = 1.09, 95% confidence interval: 1.04, 1.14), but when a more extreme group of small-for-gestational-age women (below -3 standard deviations) was defined, the association was less evident (hazard ratio = 1.04, 95% confidence interval: 0.94, 1.16). The results suggest that very-low-birth-weight women and, possibly, women born preterm or very preterm have a reduced probability of giving birth, while the results regarding small for gestational age are less clear. PMID- 15800265 TI - Mercury exposure from dental filling placement during pregnancy and low birth weight risk. AB - Several European countries have guidelines suggesting that women should not receive mercury-containing dental amalgam fillings during pregnancy. One concern raised by several studies is that mercury exposure during pregnancy may lead to decreased birth weight. A population-based, case-control study was designed to investigate whether placement of mercury-containing fillings in 1993-2000 during pregnancy increased the low-birth-weight risk. Cases and controls were sampled from enrollees of a dental insurance plan with live singleton births in Washington State; 1,117 women with low-birth-weight infants (< 2,500 g) were compared with a random sample of 4,468 women with infants weighing 2,500 g or more. The results indicated that 13% of a dentally insured population had one or more restorative procedures during pregnancy that, regardless of chemical composition, did not increase the low-birth-weight risk (odds ratio = 0.96, 95% confidence interval: 0.88, 1.05). The 4.9% of the women (n = 249) who had at least one mercury-containing amalgam filling during pregnancy were not at an increased risk for a low-birth-weight infant (odds ratio = 0.75, 95% confidence interval: 0.45, 1.26) and neither were women who had 4-11 amalgam fillings placed (odds ratio = 1.00, 95% confidence interval: 0.27, 3.68). This study found no evidence that mercury-containing dental fillings placed during pregnancy increased low-birth-weight risk. PMID- 15800266 TI - Reproductive and menstrual risk factors for pancreatic cancer: a population-based study of San Francisco Bay Area women. AB - Study results regarding risks associated with reproductive characteristics and pancreatic cancer have been mixed. Using data from a population-based case control study of pancreatic cancer, the authors assessed the role of menstrual factors, reproductive factors, and hormone use in the etiology of pancreatic cancer among women (241 cases, 818 controls). Rapid case ascertainment was used in six San Francisco Bay Area counties in California between 1995 and 1999. Controls were sampled by using random digit dialing. All statistical tests were two sided. Age at menopause (> or = 45 years vs. < 45 years) was associated with a 1.8-fold increased risk of pancreatic cancer (95% confidence interval: 1.2, 2.8). No association was found between age at menarche, parity, oral contraceptive use, estrogen replacement therapy (ERT), or history of oophorectomy and pancreatic cancer. The adjusted odds ratio for current smoking and pancreatic cancer was stronger for women who had never used oral contraceptives or ERT (odds ratio = 11.5, 95% confidence interval: 3.5, 38.1) than for those who reported using both (odds ratio = 1.7, 95% confidence interval: 0.56, 5.0). Other than a possible reduced risk estimate for smoking-related pancreatic cancer for users of exogenous hormones (oral contraceptives and ERT), results did not show a consistent pattern for reproductive factors, estrogen exposure, and risk of pancreatic cancer. PMID- 15800267 TI - Repeated occurrence of basal cell carcinoma of the skin and multifailure survival analysis: follow-up data from the Nambour Skin Cancer Prevention Trial. AB - The aim of this study was to apply multifailure survival methods to analyze time to multiple occurrences of basal cell carcinoma (BCC). Data from 4.5 years of follow-up in a randomized controlled trial, the Nambour Skin Cancer Prevention Trial (1992-1996), to evaluate skin cancer prevention were used to assess the influence of sunscreen application on the time to first BCC and the time to subsequent BCCs. Three different approaches of time to ordered multiple events were applied and compared: the Andersen-Gill, Wei-Lin-Weissfeld, and Prentice Williams-Peterson models. Robust variance estimation approaches were used for all multifailure survival models. Sunscreen treatment was not associated with time to first occurrence of a BCC (hazard ratio = 1.04, 95% confidence interval: 0.79, 1.45). Time to subsequent BCC tumors using the Andersen-Gill model resulted in a lower estimated hazard among the daily sunscreen application group, although statistical significance was not reached (hazard ratio = 0.82, 95% confidence interval: 0.59, 1.15). Similarly, both the Wei-Lin-Weissfeld marginal-hazards and the Prentice-Williams-Peterson gap-time models revealed trends toward a lower risk of subsequent BCC tumors among the sunscreen intervention group. These results demonstrate the importance of conducting multiple-event analysis for recurring events, as risk factors for a single event may differ from those where repeated events are considered. PMID- 15800268 TI - Intakes of calcium and vitamin D and risk of colorectal cancer in women. AB - In vivo and in vitro studies have suggested a protective role of calcium and vitamin D in the development of colorectal cancer. However, epidemiologic data have been inconclusive. The authors prospectively assessed intakes of calcium and vitamin D in relation to risk of colorectal cancer in a large, prospective, female cohort from the US Women's Health Study. In 1993, 39,876 women aged > or = 45 years and free of cardiovascular disease and cancer were enrolled in the study. During an average follow-up of 10 years, 223 of 36,976 women eligible for the present study developed colorectal cancer. Intakes of calcium and vitamin D from dietary sources and supplements were assessed with a baseline food frequency questionnaire. Cox proportional hazards regression was used to estimate relative risks and 95% confidence intervals. Intakes of total calcium and vitamin D were not associated with risk of colorectal cancer; multivariate relative risks comparing the highest with the lowest quintile were 1.20 (95% confidence interval: 0.79, 1.85; p for trend = 0.21) for total calcium and 1.34 (95% confidence interval: 0.84, 2.13; p for trend = 0.08) for total vitamin D. Intakes of both nutrients from specific types of sources, including diet and supplements, were also not significantly associated with colorectal cancer risk. Data provide little support for an association of calcium and vitamin D intake with colorectal cancer risk. PMID- 15800269 TI - Application of the case-crossover design to reduce unmeasured confounding in studies of condom effectiveness. AB - This analysis examined how unmeasured confounding affects estimates of the effectiveness of condoms in preventing sexually transmitted infections. Data were analyzed from a prospective cohort study of 1,122 female sexually transmitted disease clinic patients in Alabama (1992-1995), wherein participants were evaluated for sexually transmitted infections at six 1-month intervals. Associations between condom use and incident gonorrhea and chlamydia infection were compared between case-crossover and cohort analyses. In a case-crossover analysis of 228 follow-up visits ending in gonorrhea/chlamydia ("case intervals") and 743 self-matched follow-up visits not ending in gonorrhea/chlamydia ("noncase intervals") (183 women), consistent condom use without breakage or slippage was associated with significantly reduced risk of infection relative to nonuse (adjusted risk odds ratio = 0.49, 95% confidence interval: 0.26, 0.92). Conversely, a cohort analysis of 245 case intervals and 3,896 noncase intervals (919 women) revealed no significant reduction in infection risk from consistent use of condoms (adjusted risk odds ratio = 0.79, 95% confidence interval: 0.53, 1.17). Dose-response relations between the number of unprotected sex acts and infection were stronger in the case-crossover analysis (p for trend = 0.009) than in the cohort analysis (p for trend = 0.18). These findings suggest that epidemiologic studies confounded by unmeasured differences between condom users and nonusers underestimate condom effectiveness against these infections. The case-crossover method provides an additional technique for reducing unmeasured confounding in studies of condom effectiveness. PMID- 15800270 TI - Young age at first sexual intercourse and sexually transmitted infections in adolescents and young adults. AB - The authors examined the relation between age at first vaginal intercourse and a positive nucleic acid amplification test for sexually transmitted infection (STI). A nationally representative sample of 9,844 respondents aged 18-26 years was tested for chlamydial infection, gonorrhea, and trichomoniasis in wave 3 (2001-2002) of the National Longitudinal Study of Adolescent Health. The authors used multiple logistic regression to assess the relation between age at first sexual intercourse and these STIs and to examine variation by current age, sex, race, and ethnicity. Younger ages at first intercourse were associated with higher odds of STI in comparison with older ages, but the effect diminished with increasing current age. For example, the odds of having an STI for an 18-year-old who first had intercourse at age 13 were more than twice those of an 18-year-old who first had intercourse at age 17 (prevalence odds ratio = 2.25, 95% confidence interval: 1.42, 3.59). In contrast, the odds of having an STI among 24-year-olds with first intercourse at age 13 versus those with first intercourse at age 17 were the same (prevalence odds ratio = 1.11, 95% confidence interval: 0.88, 1.39). Thus, earlier initiation of sexual intercourse is strongly associated with STIs for older adolescents but not for young adults over age 23 years. PMID- 15800271 TI - Substantial overestimation of standard errors of relative survival rates of cancer patients. AB - Relative survival rates are among the most commonly reported outcome measures of cancer patients. They are calculated as ratios of observed survival rates and the expected survival rates in the absence of cancer. Standard errors of relative survival rates are commonly calculated by dividing the standard error for absolute survival rates by the expected survival, without taking possible random variation of the latter into account. The aim of this study was to empirically assess the validity of these commonly reported standard errors. Using data from the nationwide Finnish Cancer Registry, the authors calculated 5- and 10-year absolute, expected, and relative survival rates for patients with 25 common forms of cancer in Finland in 1989. The authors used bootstrap analysis to empirically assess the random error of absolute and relative survival rates and then compared the results with conventionally derived estimates of standard errors. The conventional and bootstrap standard errors were closely similar for all estimates of absolute survival. By contrast, the conventional estimates of standard errors of 5- and 10-year relative survival exceeded the bootstrap estimates by up to 17% and 32%, respectively. The authors conclude that conventional derivation may substantially overestimate standard errors for relative survival. PMID- 15800272 TI - Improving population attributable fraction methods: examining smoking attributable mortality for 87 geographic regions in Canada. AB - Smoking-attributable mortality (SAM) is the number of deaths in a population caused by smoking. In this study, the authors examined and empirically quantified the effects of methodological problems in the estimation of SAM through population attributable fraction methods. In addition to exploring common concerns regarding generalizability and residual confounding in relative risks, the authors considered errors in measuring estimates of risk exposure prevalence and mortality in target populations and estimates of relative risks from etiologic studies. They also considered errors resulting from combining these three sources of data. By modifying SAM estimates calculated using smoking prevalence obtained from the 2000-2001 Canadian Community Health Survey, a population-based survey of 131,535 Canadian households, the authors observed the following effects of potential errors on estimated national SAM (and the range of effects on 87 regional SAMs): 1) using a slightly biased, mismatched definition of former smoking: 5.3% (range, 1.8% to 11.6%); 2) using age-collapsed prevalence and relative risks: 6.9% (range, 1.1% to 15.5%) and -15.4% (range, -7.9% to 21.0%), respectively; 3) using relative risks derived from the same cohort but with a shorter follow-up period: 8.7% (range, 4.5% to 11.8%); 4) using relative risks for all diseases with age-collapsed prevalence: 49.7% (range, 24.1% to 82.2%); and 5) using prevalence estimates unadjusted for exposure-outcome lag: 14.5% (range, -20.8% to 42.6%) to -1.4% (range, -0.8% to -2.7%), depending on the method of adjustment. Applications of the SAM estimation method should consider these sources of potential error. PMID- 15800273 TI - Hormone give-and-take. PMID- 15800274 TI - Feeling spunky With JNK. PMID- 15800275 TI - Mopping up nuclear waste. PMID- 15800276 TI - Evidence-based recommendations for best practices in weight loss surgery. PMID- 15800277 TI - Commonwealth of Massachusetts Betsy Lehman Center for Patient Safety andMedical Error Reduction Expert Panel on Weight Loss Surgery: executive report. PMID- 15800278 TI - Best practice recommendations for surgical care in weight loss surgery. AB - OBJECTIVE: To establish evidence-based guidelines for best practices for surgical care in weight loss surgery (WLS). RESEARCH METHODS AND PROCEDURES: We carried out a systematic search of English-language literature on WLS in MEDLINE and the Cochrane Library. Key words were used to narrow the field for a selective review of abstracts. Data extraction was performed, and evidence categories were assigned according to a grading system based on established evidence-based models. RESULTS: We assessed types of WLS, recommended guidelines for appropriateness, developed strategies for medical error reduction, established criteria for credentialing of systems and practitioners, and specified research needed for the future. DISCUSSION: Surgeon training, credentialing, and type of surgery performed were identified as key factors in patient safety. Other important issues in the delivery of best practice care included appropriate patient selection; use of a multidisciplinary treatment team; facility staffing, equipment, and administrative support; and early recognition and proper management of complications. PMID- 15800279 TI - Criteria for patient selection and multidisciplinary evaluation and treatment of the weight loss surgery patient. AB - OBJECTIVE: To provide evidence-based guidelines for patient selection and to recommend the medical and nutritional aspects of multidisciplinary care required to minimize perioperative and postoperative risks in patients with severe obesity who undergo weight loss surgery (WLS). RESEARCH METHODS AND PROCEDURES: Members of the Multidisciplinary Care Task Group conducted searches of MEDLINE and PubMed for articles related to WLS in general and medical and nutritional care in particular. Pertinent abstracts and literature were reviewed for references. Multiple searches were carried out for various aspects of multidisciplinary care published between 1980 and 2004. A total of 3000 abstracts were identified; 242 were reviewed in detail. RESULTS: We recommended multidisciplinary screening of WLS patients to ensure appropriate selection; preoperative assessment for cardiovascular, pulmonary, gastrointestinal, endocrine, and other obesity-related diseases associated with increased risk for complications or mortality; preoperative weight loss and cessation of smoking; perioperative prophylaxis for deep vein thrombosis and pulmonary embolism (PE); preoperative and postoperative education and counseling by a registered dietitian; and a well-defined postsurgical diet progression. DISCUSSION: Obesity-related diseases are often undiagnosed before WLS, putting patients at increased risk for complications and/or early mortality. Multidisciplinary assessment and care to minimize short- and long-term risks include: comprehensive medical screening; appropriate pre-, peri-, and postoperative preparation; collaboration with multiple patient care disciplines (e.g., anesthesiology, pulmonary medicine, cardiology, and psychology); and long-term nutrition education/counseling. PMID- 15800280 TI - Behavioral and psychological factors in the assessment and treatment of obesity surgery patients. AB - OBJECTIVE: To provide evidence-based guidelines on the psychological and behavioral screening of weight loss surgery (WLS) candidates and the impact of psychosocial factors on behavior change after gastric bypass surgery. RESEARCH METHODS AND PROCEDURES: The members of the Behavioral and Psychological subgroup of the Multidisciplinary Care Task Group conducted searches of MEDLINE and PubMed for articles related to WLS, behavior changes, and mental health, including quality of life (QOL) and behavior modification. Pertinent abstracts and literature were reviewed for references. A total of 198 abstracts were identified; 17 papers were reviewed in detail. Search periods were from 1980 to 2004. RESULTS: We found a high incidence of depression, negative body image, eating disorders, and low QOL in severely obese patients. Our task subgroup recommended that all WLS candidates be evaluated by a licensed mental health care provider (i.e., psychiatrist, psychologist, or social worker), experienced in the treatment of severely obese patients and working within the context of a multidisciplinary care team. We also recommended development of pre- and postsurgical treatment plans that address psychosocial contraindications for WLS and potential barriers to postoperative success. DISCUSSION: The psychological consequences of obesity can range from lowered self-esteem to clinical depression. Rates of anxiety and depression are three to four times higher among obese individuals than among their leaner peers. A comprehensive multidisciplinary program that incorporates psychological and behavior change services can be of critical benefit in enhancing compliance, outcome, and QOL in WLS patients. PMID- 15800281 TI - Best practice guidelines on informed consent for weight loss surgery patients. AB - OBJECTIVE: To provide evidence-based guidelines on informed consent and the education that underlies it for legally competent, severely obese weight loss surgery (WLS) patients. RESEARCH METHODS AND PROCEDURES: We conducted a systematic review of the scientific literature published on MEDLINE between 1984 and 2004. Three articles focused on informed consent for WLS; none was based on empirical studies. We summarized each paper and assigned evidence categories according to a grading system derived from established evidence-based models. We also relied on informed consent and educational materials from six WLS programs in Massachusetts. All evidence is Category D. Recommendations were based on a review of the available literature, informed consent materials from WLS programs, and expert opinion. RESULTS: This Task Group found that the informed consent process contributes to long-term outcome in multiple ways but is governed by limited legal requirements. We focused our report on the legal and ethical issues related to informed consent, i.e., disclosure vs. comprehension. Recommendations centered on the importance of assessing patient comprehension of informed consent materials, the content of those materials, and the use of active teaching/learning techniques to promote understanding. DISCUSSION: Although demonstrated comprehension is not a legal requirement for informed consent in Massachusetts or other states, the members of this Task Group found that the best interests of WLS patients, providers, and facilities are served when clinicians engage patients in active learning and collaborative decision making. PMID- 15800282 TI - Best practice recommendations for anesthetic perioperative care and pain management in weight loss surgery. AB - OBJECTIVE: To develop evidence-based recommendations that optimize the safety and efficacy of perioperative anesthetic care and pain management in weight loss surgery (WLS) patients. RESEARCH METHODS AND PROCEDURES: This Task Group examined the scientific literature on anesthetic perioperative care and pain management published in MEDLINE from January 1994 to March 2004. We also reviewed additional data from other sources (e.g., book chapters). The search yielded 195 abstracts, of which 35 references were reviewed in detail. Task Group consensus was used to provide recommendations when evidence in the literature was insufficient. RESULTS: We developed anesthesia practice and patient safety advisory recommendations for preoperative evaluation, intraoperative management, and postoperative care and pain management of WLS patients. We also provided suggestions related to medical error reduction and systems improvements, credentialing, and future research. DISCUSSION: Obesity-related comorbidities including obstructive sleep apnea place WLS patients at increased risk for complications perioperatively. Regarding perioperative safety and outcomes, conclusive evidence beyond the accepted standard of care in the reviewed literature is limited. Few reports specifically address the perioperative needs of severely obese patients. In this advisory, we synthesize current knowledge and make best practice recommendations for perioperative care and pain management in WLS patients. These recommendations require periodic review as further medical knowledge and evidence evolve. PMID- 15800283 TI - Best practices for perioperative nursing care for weight loss surgery patients. AB - OBJECTIVE: To describe the unique nursing responsibilities involved in providing nursing care to severely obese weight loss surgery patients and to develop evidence-based guidelines for safe patient care. RESEARCH METHODS AND PROCEDURES: We performed a systematic review of the scientific literature using MEDLINE and CINAHL. A specific search of nursing journals from 1985 to 2004 identified 134 articles; 16 were found to be pertinent. These were reviewed in detail and used in the context of this report. The quality of the evidence was graded according to a system derived from established evidence-based models. Recommendations were developed from published evidence and expert opinion. RESULTS: This Task Group found that safe and competent nursing care requires assessment of, and provision for, the complex physical and psychological needs of weight loss surgery patients. We developed evidence-based guidelines for preoperative, perioperative, and postoperative care that address risk factors unique to severely obese patients. We also addressed issues related to the use of proper body mechanics and positioning to avoid on-the-job injury to nursing staff. DISCUSSION: We found that patient safety is best served when nurses are specifically trained to deal with the physical, medical, and psychosocial needs of severely obese patients and when they play an integral role in the multidisciplinary healthcare team. This role should start with a patient's first contact with the system and continue through discharge and follow-up. Special attention needs to be paid to the widespread bias and discrimination that severely obese individuals often experience. PMID- 15800284 TI - Best practice guidelines in pediatric/adolescent weight loss surgery. AB - OBJECTIVE: To establish evidence-based guidelines for best practices in pediatric/adolescent weight loss surgery (WLS). RESEARCH METHODS AND PROCEDURES: We carried out a systematic search of English-language literature in MEDLINE on WLS performed on children and adolescents. Key words were used to narrow the field for a selective review of abstracts. Data were extracted, and evidence categories were assigned according to a grading system based on established evidence-based models. Eight pertinent case series, published between 1980 and 2004, were identified and reviewed. These data were supplemented with expert opinions and literature on WLS in adults. RESULTS: Recommendations focused on patient safety, reduction of medical errors, systems improvements, credentialing, and future research. We developed evidence-based criteria for eligibility, assessment, treatment, and follow-up; recommended surgical procedures based on the best available evidence; and established minimum guideline requirements for data collection. DISCUSSION: Lack of adequate data and gaps in knowledge were cited as important reasons for caution. Physiological status, comprehensive screening of patients and their families, and required education and counseling were identified as key factors in assessing eligibility for surgery. Data collection and peer review were also identified as important issues in the delivery of best practice care. PMID- 15800285 TI - Specialized staff and equipment for weight loss surgery patients: best practice guidelines. AB - OBJECTIVE: To provide evidence-based guidelines on the specialized personnel, equipment, and physical plant required for safe and effective care of severely obese weight loss surgery (WLS) patients. RESEARCH METHODS AND PROCEDURES: We examined MEDLINE (Ovid and PubMed) and the Cumulative Index of Nursing and Allied Health Literature for articles on facilities resources for care of WLS patients published in English between January 1980 and March 2004. We queried several web sites for appropriate references; these included the Agency for Healthcare Research and Quality and the American College of Surgeons. The majority of reference material was descriptive and not specific to facilities resources for WLS patients. We identified a substantial body of literature on the general subject of patient safety; three of these articles were used to develop recommendations on the use of technology for medical error reduction. All other recommendations are based on 11 expert opinion reports. RESULTS: We recommended adequate training and credentialing for all medical staff; dedicated support and administrative personnel; and specialized interventional, diagnostic, operating room, and transport equipment. We specified needed adaptations to the physical plant and developed evidence-based guidelines for medical error reduction and systems improvements. DISCUSSION: Specialized resources and dedicated staff are needed to protect the health of WLS surgery patients and staff. Adaptations include preoperative preparation for safe means of patient transport; techniques of anesthesia and intraoperative exposure; provisions for postoperative recovery; and measures to assure postoperative patient safety, hygiene, and comfort. PMID- 15800286 TI - Coding and reimbursement for weight loss surgery: best practice recommendations. AB - OBJECTIVE: To review the use and usefulness of billing codes for services related to weight loss surgery (WLS) and to examine third party reimbursement policies for these services. RESEARCH METHODS AND PROCEDURES: The Task Group carried out a systematic search of MEDLINE, the Internet, and the trade press for publications on WLS, coding, reimbursement, and coding and reimbursement policy. Twenty-eight articles were each reviewed and graded using a system based on established evidence-based models. The Massachusetts Dietetics Association provided reimbursement data for nutrition services. Three suppliers of laparoscopic WLS equipment provided summaries of coding and reimbursement information. WLS program directors were surveyed for information on use of procedure codes related to WLS. RESULTS: Recommendations focused on correcting or improving on the current lack of congruity among coding practices, reimbursement policies, and accepted clinical practice; lack of uniform coding and reimbursement data across institutions; inconsistent and/or inaccurate diagnostic and billing codes; inconsistent insurance reimbursement criteria; and inability to leverage reimbursement and coding data to track outcomes, identify best practices, and perform accurate risk-benefit analyses. DISCUSSION: Rapid changes in the prevalence of obesity, our understanding of its clinical impact, and the technologies for surgical treatment have yet to be adequately reflected in coding, coverage, and reimbursement policies. Issues identified as key to effective change include improved characterization of the risks, benefits, and costs of WLS; anticipation and monitoring of technological advances; encouragement of consistent patterns of insurance coverage; and promotion of billing codes for WLS procedures that facilitate accurate tracking of clinical use and outcomes. PMID- 15800287 TI - Data collection systems for weight loss surgery: an evidence-based assessment. AB - OBJECTIVE: To evaluate the existence and efficacy of data collection systems for weight loss surgery (WLS) and establish evidence-based guidelines for the development of a statewide WLS registry in Massachusetts. RESEARCH METHODS AND PROCEDURES: We conducted two systematic searches of English language literature in MEDLINE. The first was on data collection registries related to WLS; the second was an expanded search encompassing other surgical fields (e.g., cardiac and thoracic surgery) and registries (i.e., cancer). Fourteen articles were found to be pertinent. Data were extracted, and evidence categories were assigned according to a grading system based on established evidence-based models. Recommendations were derived from these literature reviews and expert opinion. RESULTS: This task group found that there are no standardized data collection systems for WLS in Massachusetts (or any other states) and no mandated reporting of WLS-specific outcomes. We described existing WLS databases and systems in other surgical fields. Recommendations focused on the importance and feasibility of data collection for WLS and the need to conduct a pilot study and explore options for creating a statewide WLS database. DISCUSSION: A statewide WLS data collection system would optimize patient care by enabling the collection, analysis, and dissemination of best practice data. A broad-based effort is needed to meet challenges involved in defining and implementing such a system. PMID- 15800288 TI - Changes in insulin secretion and glucose metabolism induced by dexamethasone in lean and obese females. AB - OBJECTIVE: In healthy lean individuals, changes in insulin sensitivity occurring as a consequence of a 2-day dexamethasone administration are compensated for by changes in insulin secretion, allowing glucose homeostasis to be maintained. This study evaluated the changes in glucose metabolism and insulin secretion induced by short-term dexamethasone administration in obese women. RESEARCH METHODS AND PROCEDURES: Eleven obese women with normal glucose tolerance were studied on two occasions, without and after 2 days of low-dose dexamethasone administration. A two-step hyperglycemic clamp (7.5 and 10 mM glucose) with 6,6 (2)H(2) glucose was used to assess insulin secretion and whole body glucose metabolism. Results were compared with those obtained in a group of eight lean women. RESULTS: Without dexamethasone, obese women had higher plasma insulin concentrations in the fasting state, during the first phase of insulin secretion, and at the two hyperglycemic plateaus. However, they had normal whole body glucose metabolism compared with lean women, indicating adequate compensation. After dexamethasone, obese women had a 66% to 92% increase in plasma insulin concentrations but a 15.4% decrease in whole body glucose disposal. This contrasted with lean women, who had a 91% to 113% increase in plasma insulin concentrations, with no change in whole body glucose disposal. DISCUSSION: Dexamethasone administration led to a significant reduction in whole body glucose disposal at fixed glycemia in obese but not lean women. This indicates that obese women are unable to increase their insulin secretion appropriately. PMID- 15800289 TI - Heritability of body composition measured by DXA in the diabetes heart study. AB - OBJECTIVE: The purpose of this study was to investigate the heritability of body composition measured by DXA in the Diabetes Heart Study (DHS). RESEARCH METHODS AND PROCEDURES: Participants were 292 women and 262 men (age, 38 to 86 years; BMI, 17 to 57 kg/m(2)) from 244 families. There were 492 white and 49 African American sibling pairs. DXA measurements of percentage fat mass (FM), whole body FM, and lean mass (LM), as well as regional measurements of trunk fat mass (TFM) and appendicular lean mass (ALM), were obtained. Heritability of FM, LM, and BMI were estimated using Sequential Oligogenic Linkage Analysis Routines. RESULTS: After adjusting for age, gender, ethnicity, and height, the heritability estimates of various compositional attributes were %FM = 0.64, whole body FM = 0.71, TFM = 0.63, whole body LM = 0.60, ALM = 0.66, and BMI = 0.64 (all p < 0.0001). Additional adjustment for diabetes status, smoking, dietary intake, and physical activity resulted in only minor changes in the heritability estimates (h(2) = 0.63 to 0.72, all p < 0.0001). Furthermore, heritability of TFM after additional adjustment for whole body FM was significant (h(2) = 0.55, p < 0.0001), and heritability of ALM after additional adjustment for whole body LM was also significant (h(2) = 0.51, p < 0.0001). DISCUSSION: These data suggest that FM and LM measured by DXA are highly heritable and can be effectively used in designing linkage studies to locate genes governing body composition. In addition, regional distribution of FM and LM may be genetically determined. PMID- 15800290 TI - Depressive mood and abdominal fat distribution in overweight premenopausal women. AB - OBJECTIVE: There is increasing evidence that depressive mood is associated with central obesity, but little is known about the association between depression and abdominal fat distribution. This study investigated this relationship in premenopausal women. RESEARCH METHODS AND PROCEDURES: We recruited 101 overweight premenopausal women who had no eating disorders as defined using the DSM IV criteria. Depressive mood was assessed using Zung's Self-Rating Depression Scale (SDS). Areas of visceral (VAT) and subcutaneous (SAT) adipose tissue at the level of vertebral body L(4)-L(5) were measured using computed tomography. Associations of VAT, SAT, and the ratio of VAT to SAT with natural logarithmic transformation [(ln)]SDS were evaluated using linear regression. Anthropometric indices and physical fitness were also measured. Information on socioeconomic status, education level, and alcohol and smoking habits was obtained using self administered questionnaires. A hospital nutritionist assessed nutritional status. All of these factors were adjusted for as possible confounding factors in the analyses. RESULTS: The (ln)SDS score showed a positive association with the area of VAT, even after adjusting for the confounders mentioned above (p < 0.01). BMI, waist circumference, maximal oxygen uptake, and age were also associated with the area of VAT (all p < 0.05). In contrast, the (ln)SDS score was not associated with SAT (p > 0.10). DISCUSSION: We showed that depressive mood is associated with VAT, not with SAT, in overweight premenopausal women. These findings may explain some of the association between depression and coronary heart disease. More studies are needed to elucidate the causal relationship. PMID- 15800291 TI - Pediatricians' own weight: self-perception, misclassification, and ease of counseling. AB - OBJECTIVES: Pediatricians underdiagnose overweight and feel ineffective at counseling. Given the relationship between physicians' health and health habits and counseling behaviors, we sought to determine the 1) percentage of pediatricians who are overweight; 2) accuracy of pediatricians' own weight status classification; and 3) relationship between weight self-perception and perceived ease of obesity counseling. RESEARCH METHODS AND PROCEDURES: This study was a cross-sectional, mail survey of North Carolina pediatricians that queried about their weight status and ease of counseling. Accuracy of pediatricians' self classification of weight status was compared with BMIs derived from self-reported height and weight. Using logistic regression, controlling for potential confounding variables, we examined the association between weight perception and ease of counseling. RESULTS: The unadjusted response rate was 62%, and the adjusted response rate was 71% (n = 355). Nearly one-half (49%) of overweight pediatricians did not identify themselves as such. Men had greater adjusted odds of misclassifying overweight than women [odds ratio (OR), 3.61; 95% confidence interval (CI) = 1.81, 7.21]. Self-classified "thin" pediatricians had nearly six times the odds of reporting more counseling difficulty as a result of their weight than "average" weight pediatricians (OR = 5.69; 95% CI = 2.30, 14.1), and self-identified "overweight" pediatricians reported nearly four times as great counseling difficulty as "average" weight physicians (OR = 3.84; 95% CI = 1.11, 13.3), after adjustment for self-reported BMI weight status and other potential confounders. DISCUSSION: The roles that physician weight misclassification and self-perception potentially play in influencing rates of obesity counseling warrant further research. PMID- 15800292 TI - Prevalence of weight cycling and its relation to health indicators in Finland. AB - OBJECTIVE: This study was performed to identify weight cyclers and to assess the prevalence of weight cycling and its relation to health indicators in Finnish adults. RESEARCH METHODS AND PROCEDURES: Data for the study consisted of 3320 men and 3540 women (25 to 64 years of age). The subjects went through a health examination, and data on intentional weight losses and regains during the last 10 years were collected by a questionnaire. The subjects were divided into five groups: severe weight cyclers (weight loss >/= 5 kg at least three times with regain), mild weight cyclers (weight loss >/= 5 kg one to two times and regain), successful dieters (weight loss >/= 5 kg with no regain), nonobese nondieters, and obese nondieters. RESULTS: Approximately 7% of men and 10% of women were defined as severe weight cyclers, and an additional 11% and 19% were defined as mild weight cyclers, respectively. In men, 20% of severe weight cyclers and 15% of obese nondieters perceived their health as poor, whereas in other groups, these proportions were <10%. In both sexes, severe weight cyclers seemed to have visited a doctor more frequently than nonobese nondieters. Severe weight cyclers were also more likely to use some medication compared with other groups. DISCUSSION: Weight cycling is more common in women than in men, and it seems to be associated with more regular visits to a doctor and poor self-perceived health. However, because of the cross-sectional design of the study, causal conclusions cannot be drawn. PMID- 15800293 TI - Obesity among people with and without mental retardation across adulthood. AB - OBJECTIVE: This study was designed to explore obesity during adulthood and the likelihood of moving out of obesity among 1809 adults without disability and 680 adults with mental retardation who received care at the same primary care practices during the period of 1990 to 2003. RESEARCH METHOD AND PROCEDURES: A retrospective observational design using medical records first identified patients with mental retardation (MR) and age-matched controls without disabilities. Data on BMI collected during each primary care visit allowed exploration of obesity at three levels. Moving out of obesity was defined as having a BMI <25 kg/m(2). We also abstracted data on age, sex, race, and other medical conditions. RESULTS: For adults 20 to 29 years of age, 33.1% of patients without disability and 21% of patients with MR had a BMI >30 kg/m(2). Between the ages of 50 and 59 years, 40.5% of the patients without disability and 35.2% of the patients with MR had a BMI >30 kg/m(2). Patients with mild MR had similar prevalence rates of obesity and patients with severe MR had significantly lower prevalence of obesity compared with the patients without disability through 50 years of age. Throughout the period from 20 to 60 years of age, between 15% and 40% of individuals with and without MR, who were previously obese, were not currently obese. DISCUSSION: Throughout the adult years, an increasing proportion of individuals with and without MR are obese. However, obesity is not a chronic state; many people transition back to a normal body weight. PMID- 15800294 TI - Knee structural alteration and BMI: a cross-sectional study. AB - OBJECTIVE: To describe the associations among BMI, knee cartilage morphology, and bone size in adults. RESEARCH METHODS AND PROCEDURES: A cross-sectional convenience sample of 372 male and female subjects (mean age, 45 years; range, 26 to 61 years) was studied. Knee articular cartilage defect score (0 to 4) and prevalence (defect score of >/=2), volume, and thickness, as well as bone surface area and/or volume, were determined at the patellar, tibial, and femoral sites using T1-weighted fat-saturation magnetic resonance imaging. Height, weight, BMI, and radiographic osteoarthritis were measured by standard protocols. RESULTS: In multivariate analysis in the whole group, BMI was significantly associated with knee cartilage defect scores (beta: +0.016/kg/m(2) to +0.083/kg/m(2), all p < 0.05) and prevalence (odds ratio: 1.05 to 1.12/kg/m(2), all p < 0.05 except for the lateral tibiofemoral compartment). In addition, BMI was negatively associated with patellar cartilage thickness only (beta = -0.021 mm/kg/m(2); p = 0.039) and was positively associated with tibial bone area (medial: beta = +7.1 mm(2)/kg/m(2), p = 0.001; lateral: beta = +3.2 mm(2)/kg/m(2), p = 0.037). Those who were obese also had higher knee cartilage defect severity and prevalence and larger medial tibial bone area but no significant change in cartilage volume or thickness compared with those of normal weight. DISCUSSION: This study suggests that knee cartilage defects and tibial bone enlargement are the main structural changes associated with increasing BMI particularly in women. Preventing these changes may prevent knee osteoarthritis in overweight and obese subjects. PMID- 15800295 TI - Additive interactions of maternal prepregnancy BMI and breast-feeding on childhood overweight. AB - OBJECTIVE: To examine the interactions of maternal prepregnancy BMI and breast feeding on the risk of overweight among children 2 to 14 years of age. RESEARCH METHODS AND PROCEDURES: The 1996 National Longitudinal Survey of Youth, Child and Young Adult data in the United States were analyzed (n = 2636). The weighted sample represented 51.3% boys, 78.0% whites, 15.0% blacks, and 7.0% Hispanics. Childhood overweight was defined as BMI >/=95th percentile for age and sex. Maternal prepregnancy obesity was determined as BMI >/=30 kg/m(2). The duration of breast-feeding was measured as the weeks of age from birth when breast-feeding ended. RESULTS: After adjusting for potential confounders, children whose mothers were obese before pregnancy were at a greater risk of becoming overweight [adjusted odds ratio (OR), 4.1; 95% confidence interval (CI), 2.6, 6.4] than children whose mothers had normal BMI (<25 kg/m(2); p < 0.001 for linear trend). Breast-feeding for >/=4 months was associated with a lower risk of childhood overweight (OR, 0.6; 95% CI, 0.4, 1.0; p = 0.06 for linear trend). The additive interaction between maternal prepregnancy obesity and lack of breast-feeding was detected (p < 0.05), such that children whose mothers were obese and who were never breast-fed had the greatest risk of becoming overweight (OR, 6.1; 95% CI, 2.9, 13.1). DISCUSSION: The combination of maternal prepregnancy obesity and lack of breast-feeding may be associated with a greater risk of childhood overweight. Special attention may be needed for children with obese mothers and lack of breast-feeding in developing childhood obesity intervention programs. PMID- 15800296 TI - Obesity and the use of health care services. AB - OBJECTIVE: This study investigated differences in the use of health care services and associated costs between obese and nonobese patients. RESEARCH METHODS AND PROCEDURES: New adult patients (N = 509) were randomly assigned to primary care physicians at a university medical center. Their use of medical services and related charges was monitored for 1 year. Data collected included sociodemographics, self-reported health status using the Medical Outcomes Study Short Form-36, evaluation for depression using the Beck Depression Index, and measured height and weight to calculate BMI. RESULTS: Obese patients included a significantly higher percentage of women and had higher mean age, lower mean education, lower mean health status, and higher mean Beck Depression Index scores. Obese patients had a significantly higher mean number of visits to both primary care (p = 0.0005) and specialty care clinics (p = 0.0006), and a higher mean number of diagnostic services (p < 0.0001). Obese patients also had significantly higher primary care (p = 0.0058), specialty clinic (p = 0.0062), emergency department (p = 0.0484), hospitalization (p = 0.0485), diagnostic services (p = 0.0021), and total charges (p = 0.0033). Controlling for health status, depression, age, education, income, and sex, obesity was significantly related to the use of primary care (p = 0.0364) and diagnostic services (p = 0.0075). There was no statistically significant relationship between obesity and medical expenditures in any of the five categories or for total charges. DISCUSSION: Obesity is a chronic condition requiring long-term management, with an emphasis on prevention. If this critical health issue is not appropriately addressed, the prevalence of obesity and obesity-related diseases will continue to grow, resulting in escalating use of health care services. PMID- 15800297 TI - The importance of cognitive outcomes. PMID- 15800298 TI - Ubiquitous but unfinished: online information retrieval systems. PMID- 15800299 TI - The value of diagnostic information to patients with chest pain suggestive of coronary artery disease. AB - BACKGROUND: The results of diagnostic tests often have a profound impact on the way patients view their health. Decision analyses and clinical guidelines do not routinely consider this, in part because methods are not well developed for measuring the value of testing to patients. OBJECTIVES: To measure the value of stress testing to patients with chest pain suggestive of coronary artery disease (CAD) and to improve methods for measuring the value of diagnostic information. METHODS: The authors conducted a prospective cohort study of patients with chest pain who were referred from 44 primary care practices for treadmill testing (N = 320). Current health status, perceived life expectancy, anxiety, uncertainty, and preferences for current health states were measured before and 1 week after testing and receipt of the results. Patients also reported the diagnosis given by their physicians after testing. The authors used paired t tests to assess changes before and after testing. RESULTS: Perceived life expectancy lengthened, anxiety decreased, and uncertainty decreased 1 week after exercise testing, compared to before (P < 0.01). For many patients, sytoms were less bothersome after testing than before. There were few changes in perceptions of current health status, as measured by the SF-36. The authors found evidence of reassurance among patients who reported that CAD had been excluded and no evidence of psychological harm among patients who reported a new CAD diagnosis. CONCLUSIONS: Patients experienced measurable psychological benefits from noninvasive diagnostic testing for CAD. Similar measurements should be standard components of diagnostic test evaluation. PMID- 15800300 TI - Invariance and inconsistency in utility ratings. AB - PURPOSE: To assess utilities of composite health states for dependence in activities of daily living (ADLs) for invariance (i.e., when subjects provide a utility of 1 for all health states) and order inconsistency (i.e., when subjects order their utilities such that their utility for a combination of ADL dependencies is greater than their utility for any subset of the combination). METHODS: Each of the 400 subjects, age 65 y and older, enrolled in one of several regional medical centers of the Kaiser Permanente Medical Care Program of Northern California and provided standard-gamble utilities for single ADL dependencies (e.g., bathing, dressing, continence) and for dependence in 8 other combinations of ADL dependencies. For order-inconsistent responses, the authors calculated the maximum magnitude of inconsistency as the maximum difference between the utility for the combined ADL dependence health state and that of its inconsistent subset. RESULTS: A total of 76 subjects (19%) gave a utility of 1.0 for all health states presented to them; 19 (5%) gave the same utility other than 1.0 for all health states; 130 (33%) gave at least 1 utility < 1.0 and had no order inconsistencies; and 175 (44%) had at least 1 order inconsistency. Invariance was associated with a Mini-Mental Status Examination score < 28.6 (P = 0.01), with education < 12 y (P = 0.004), with race/ethnicity other than non Hispanic White/Caucasian (P = 0.001), and with shorter time spent on the utility elicitation task (P < 0.0001). Among the inconsistent subjects, 69% had a maximal magnitude of inconsistency that was within 1 standard deviation of the mean utilities. The maximal magnitude of inconsistency was associated with longer time spent on the elicitation task (P < 0.0001) and race/ethnicity other than non Hispanic White/Caucasian (P = 0.005). The mean (s) utility for dependence in continence among consistent subjects who were not invariant (0.88 [0.24]) was higher than among inconsistent subjects (0.80 [0.27]; P = 0.01). CONCLUSIONS: Invariance and order inconsistencies in utility ratings for complex health states occur frequently. Utilities of consistent subjects may differ from those of inconsistent subjects. Utility assessments should attempt to measure and report these patterns. PMID- 15800301 TI - A new logistic regression approach for the evaluation of diagnostic test results. AB - The value of a dichotomous diagnostic test is often described in terms of sensitivity, specificity, and likelihood ratios (LRs). Although it is known that these test characteristics vary between subgroups of patients, they are generally interpreted, on average, without considering information on patient characteristics, such as clinical signs and symptoms, or on previous test results. This article presents a reformulation of the logistic regression model that allows to calculate the LRs of diagnostic test results conditional on these covariates. The proposed method starts with estimating logistic regression models for the prior and posterior odds of disease. The regression model for the prior odds is based on patient characteristics, whereas the regression model for the posterior odds also includes the diagnostic test of interest. Following the Bayes theorem, the authors demontsrate that the regression model for the LR can be derived from taking the differences between the regression coefficients of the 2 models. In a clinical example, they demonstrate that the LRs of positive and negative test results and the sensitivity and specificity of the diagnostic test varied considerably between patients with different risk profiles, even when a constant odds ratio was assumed. The proposed logistic regression approach proves an efficient method to determine the performance of tests at the level of the individual patient risk profile and to examine the effect of patient characteristics on diagnostic test characteristics. PMID- 15800302 TI - The impact of an online evidence system on confidence in decision making in a controlled setting. AB - OBJECTIVE: To examine the impact of online evidence retrieval on clinicians' decision-making confidence and to determine if this differs for experienced doctors and nurses. METHODS: A sample of 44 doctors and 31 clinical nurse consultants (CNCs) answered 8 clinical scenarios (600 scenario answers) before and after the use of online evidence resources. Clinicians rated their confidence in scenario answers and in the evidence they found using the information system. RESULTS: Prior to using online evidence, 37% of doctors and 18% of CNCs answered the scenarios correctly. These clinicians were more confident (56% very confident or confident) in their answers than those with incorrect (34%) answers. Doctors with incorrect answers prior to searching rated their confidence significantly higher than did nurses who were incorrect. After searching, both groups answered 50% of scenarios correctly. Clinicians with correct answers had greater confidence in the evidence found compared to those with incorrect answers. Doctors were more confident in evidence found confirming an initially correct answer than were nurses. More than 50% of clinicians who persisted with an incorrect answer after searching reported that they were confident or very confident in the evidence found. Clinicians who did not know scenario answers before searching placed equal confidence in evidence that led them to a correct or incorrect answer. CONCLUSIONS: The information obtained from an online evidence system influenced clinicians' confidence in their answers to the clinical scenarios. The relationship between confidence in answers and correctness is complex. Both existing knowledge and professional role were mediating factors. The finding that many clinicians placed confidence in information that led them to incorrect answers warrants further investigation. PMID- 15800303 TI - Options for handling missing data in the Health Utilities Index Mark 3. AB - BACKGROUND: The Health Utilities Index Mark 3 (HUI3) is a tool composed of 41 questions, covering 8 attributes: vision, hearing, speech, ambulation, dexterity, emotion, cognition, and pain. Responses to these questions can define more than 972,000 health situations. This tool allows respondents to answer "Don't Know," for which there is no scoring instruction, to any given question. This situation creates a break in the scoring algorithm and leads to considerable amounts of missing data. The goal of this study is to develop strategies to deal with HUI3 scores for participants who have missing data. METHODS: The authors used data from 248 individuals enrolled in the Cataract Management Trial, focusing on the HUI3 vision and ambulation attributes, which had 19% and 10% of attribute levels missing, respectively. Inspection and deduction were used to fill in values independent of the value of the missing data, then alternative analytic techniques were compared, including mean substitution, model scoring, hot deck, multiple imputation, and regression imputation. RESULTS: Inspection and logical deduction reduced the percentage of missing information in the HUI3 by 49% to 87%. A comparison of analytic techniques used for the remaining HUI3 vision data missing demonstrated the value of building models based on internal response patterns and that simple analytic techniques fare as well as more complicated ones when the number of missing cases is small. CONCLUSION: Analyzing the pattern of responses in cases where the attribute level score is missing reduces the amount of missing data and can simplify the analytic process for the remaining missing data. PMID- 15800304 TI - A clinically based discrete-event simulation of end-stage liver disease and the organ allocation process. AB - BACKGROUND: The optimal allocation of scarce donor livers is a contentious health care issue requiring careful analysis. The objective of this article was to design a biologically based discrete-event simulation to test proposed changes in allocation policies. METHODS: The authors used data from multiple sources to simulate end-stage liver disease and the complex allocation system. To validate the model, they compared simulation output with historical data. RESULTS: Simulation outcomes were within 1% to 2% of actual results for measures such as new candidates, donated livers, and transplants by year. The model overestimated the yearly size of the waiting list by 5% in the last year of the simulation and the total number of pretransplant deaths by 10%. CONCLUSION: The authors created a discrete-event simulation model that represents the biology of end-stage liver disease and the health care organization of transplantation in the United States. PMID- 15800305 TI - Withholding or starting antibiotic treatment in patients with dementia and pneumonia: prediction of mortality with physicians' judgment of illness severity and with specific prognostic models. AB - BACKGROUND: To help decision makers plan treatment, the authors assessed clinical predictors of mortality from nursing home-acquired pneumonia in patients with dementia. METHODS: Pneumonia patients treated without (n = 165) or with antibiotics (n = 541) were enrolled in a prospective cohort study in 61 nursing homes. RESULTS: In both groups, clinical judgment of illness severity was a strong predictor for 1-week mortality. Despite large differences in frailty and mortality (83% in untreated patients and 15% in treated patients), separate multivariable logistic models included similar specific predictors. DISCUSSION: Despite profound differences between the 2 independent groups, predictors for short-term mortality were largely similar. We found that, when combined with physicians' clinical judgment, 3 readily assessed predictors (respiratory rate, fluid intake, and eating dependency) helped predict mortality. Our results, if confirmed in an independent population, can help make decision making about antibiotic treatment of pneumonia in patients with dementia more evidence-based. PMID- 15800306 TI - Medical decision making with incomplete evidence--choosing a platelet glycoprotein IIbIIIa receptor inhibitor for percutaneous coronary interventions. AB - BACKGROUND: Medical decision making must often be performed despite incomplete evidence. An example is the choice of a glycoprotein IIb/IIIa (GP2b3a) inhibitor, a class of potent antiplatelet medications, as adjunctive therapy during percutaneous coronary interventions (PCIs). GP2b3a inhibitor efficacy in reducing adverse outcomes has been well documented with multiple placebo-controlled randomized trials, but there is a paucity of comparative data about their individual equivalency. Substantial cost differentials are also present between the drugs. METHODS: A systematic review of the literature was performed to identify all randomized placebo-controlled trials of GP2b3a inhibitors as adjunctive therapy for PCI. Three complimentary methods were used to assist in decision making regarding drug equivalency. First, the data from the single direct comparative trial are analyzed from a Bayesian perspective. Next, prior information from other GP2b3a inhibitor trials in similar but not identical patient populations is incorporated. In the 3rd method, indirect comparisons of GP2b3a inhibitors are carried out using a hierarchical meta-analytic model of the placebo-controlled trials identified by the systematic review. RESULTS: A total of 12 randomized trials were identified involving 3 agents (abciximab, eptifibatide, tirofiban), but only 1 involved a direct comparison of 2 drugs (abciximab v. tirofiban). In contradiction to the original publication, the authors' Bayesian analysis both without (method 1) and with (method 2) the inclusion of some prior information suggests a reasonable probability of equivalency. The indirect comparisons from all randomized placebo-controlled trials (method 3) also failed to provide support for superiority of any agent over the others. CONCLUSION: Decision making with incomplete evidence is a difficult but frequently occurring medical dilemma. The authors propose 3 methods that may elucidate the process and illustrate them in the context of the choice of GP2b3a inhibitor for adjunctive therapy during PCI. Further data may or may not eventually lead to a different conclusion, but based on the evidence available to date, the authors' 3 methods suggest clinical equivalency between GP2b3a inhibitors, in contrast to the initial conclusions from the single comparative randomized trial. PMID- 15800307 TI - Jury rationality. PMID- 15800308 TI - Randomized trial of amifostine in locally advanced non-small-cell lung cancer patients receiving chemotherapy and hyperfractionated radiation: radiation therapy oncology group trial 98-01. AB - PURPOSE: To test the ability of the cytoprotectant, amifostine, to reduce chemoradiotherapy-induced esophagitis and evaluate its influence on quality of life (QOL) and swallowing symptoms. PATIENTS AND METHODS: A total of 243 patients with stage II to IIIA/B non-small-cell lung cancer received induction paclitaxel 225 mg/m(2) intravenously (IV) days 1 and 22 and carboplatin area under the curve (AUC) days 1 and 22, followed by concurrent weekly paclitaxel (50 mg/m(2) IV) and carboplatin (AUC 2), and hyperfractionated radiation therapy (69.6 Gy at 1.2 Gy bid). Patients were randomly assigned at registration to amifostine (AM) 500 mg IV four times per week or no AM during chemoradiotherapy. Beyond standard toxicity end points, physician dysphagia logs (PDLs), daily patient swallowing diaries, and QOL (EORTC QLQ-C30/LC-13) were also collected. Swallowing AUC analyses were calculated from patient diaries and PDLs. RESULTS: A total of 120 patients were randomly assigned to receive AM, and 122, to receive no AM (one patient was ineligible); 72% received AM per protocol or with a minor deviation. AM was associated with higher rates of acute nausea (P = .03), vomiting (P = .007), cardiovascular toxicity (P = .0001), and infection or febrile neutropenia (P = .03). The rate of >/= grade 3 esophagitis was 30% with AM versus 34% without AM (P = .9). Patient diaries demonstrated lower swallowing dysfunction AUC with amifostine (z test P = .025). QOL was not significantly different between the two arms, except for pain, which showed more clinically meaningful improvement and less deterioration at 6 weeks follow-up (v pretreatment) in the AM arm (P = .003). The median survival rates for both arms were comparable (AM, 17.3 v no AM, 17.9 months; P = .87). CONCLUSION: AM did not significantly reduce esophagitis >/= grade 3 in patients receiving hyperfractionated radiation and chemotherapy. However, patient self-assessments suggested a possible advantage to AM that is being explored with modified dosing route strategies. PMID- 15800309 TI - Phase II study of efficacy, safety, and pharmacokinetics of trastuzumab monotherapy administered on a 3-weekly schedule. AB - PURPOSE: This phase II study investigated the efficacy, safety, and pharmacokinetics of trastuzumab monotherapy given as first-line treatment once every 3 weeks (3-weekly) in women with human epidermal growth factor receptor 2 (HER2) -positive metastatic breast cancer (MBC). PATIENTS AND METHODS: Patients with previously untreated HER2-positive MBC received a loading dose of trastuzumab, 8 mg/kg intravenously (IV) and then 6 mg/kg IV at 3-week intervals until disease progression or patient withdrawal. RESULTS: In total, 105 patients received a median of five cycles of therapy (range, 1 to 35+). The overall response rate was 19% (23% in patients with measurable centrally confirmed immunohistochemistry [IHC] 3+ and/or fluorescence in situ hybridization [FISH] positive disease) and clinical benefit rate (complete and partial responses plus stable disease for at least 6 months) was 33% (36% in patients with measurable centrally confirmed IHC 3+ and/or FISH-positive disease). Median time to progression was 3.4 months (range, 0.6 to 23.6 months). The most common treatment related adverse events were rigors, pyrexia, headache, nausea, and fatigue. Median baseline left ventricular ejection fraction was 63%; this did not significantly change over the course of the study. The average exposure to trastuzumab observed in this study was similar to that in previous studies of the weekly regimen. However, as expected, mean trough trastuzumab concentrations were lower and peak levels were higher with 3-weekly trastuzumab compared with weekly treatments. CONCLUSION: Administering higher doses on a 3-weekly schedule did not compromise the efficacy and safety of trastuzumab in women with HER2-positive MBC, and average exposure was similar to that observed with weekly therapy. Three weekly trastuzumab may represent a convenient alternative to weekly administration. PMID- 15800310 TI - Postoperative adjuvant therapy with tamoxifen, tegafur plus uracil, or both in women with node-negative breast cancer: a pooled analysis of six randomized controlled trials. AB - PURPOSE: This article reports the results of a pooled analysis of six randomized trials conducted to study the efficacy of uracil and tegafur (UFT) in the adjuvant treatment of node-negative breast cancer patients. PATIENTS AND METHODS: Six randomized controlled trials on node-negative breast cancer patients were conducted from 1992 through 1995 in Japan that included the three, three-arm trials (control [no adjuvant], UFT, and tamoxifen [TAM] groups) and the three, four-arm trials (control, UFT, TAM, and UFT plus TAM groups). Pooled analysis was performed on the data obtained from these six trials (involving 2,934 patients). RESULTS: Overall survival was compared between the UFT group (including both the UFT group and the TAM plus UFT group) and the non-UFT group (control group and TAM group). A significant difference (P = .04) was observed in 5-year survival rates between the UFT (95.9%) and the non-UFT (94.0%) groups. Overall survival was also compared between the TAM group (TAM group and TAM plus UFT group) and the non-TAM group (control group plus UFT group). The 5-year survival rate (95.2%) in the TAM group was not significantly different from that (93.9%) in the non-TAM group, but the subset analysis showed a significant (P = .01) improvement in the estrogen receptor-positive subset. CONCLUSION: Adjuvant UFT improves the overall survival of node-negative breast cancer patients. Given that UFT has milder adverse effects, it is suggested that UFT can be a useful alternative to doxorubicin and cyclophosphamide, or cyclophosphamide, methotrexate, and fluorouracil in the adjuvant treatment for node-negative breast cancer. PMID- 15800311 TI - Patients with an unclassified genetic variant in the BRCA1 or BRCA2 genes show different clinical features from those with a mutation. AB - PURPOSE: To obtain and compare the probabilities of finding a mutation in the BRCA1 or BRCA2 genes, the clinical features, and the family history among patients with an unclassified variant (UV) and those with a pathogenic mutation. PATIENTS AND METHODS: The study included 70 patients: 24 with a UV (BRCA1, n = 4; BRCA2, n = 19; both, n = 1), and 46 with a mutation (BRCA1, n = 32; BRCA2, n = 14). Two of the UVs were novel variants; the rest had been reported previously as UVs. Probabilities of finding a mutation were retrospectively obtained using BRCAPRO and Myriad II programs. RESULTS: The probability to detect a mutation was significantly lower in the group of patients with a UV than in those with a mutation (BRCAPRO [mean +/- standard deviation], 0.297 +/- 0.312 v 0.627 +/- 0.315, P = .001; and Myriad II, 0.124 +/- 0.090 v 0.283 +/- 0.176, P = .001, respectively). Independent predictive factors of finding either a UV or a mutation were number of affected relatives (2.9 +/- 1.4 v 4.0 +/- 1.9; P = .039) and number of tumors among relatives (3.3 +/- 1.4 v 4.4 +/- 1.8; P = .031), respectively. CONCLUSION: The combined data about the predictive models show significant differences between both groups. Individual probabilities can be regarded as a help to guide the clinical management of patients with a UV in those genes. However, a definitive conclusion about the pathogenicity of a UV can not be obtained from the clinical features alone, but only in combination with biochemical and epidemiologic data. PMID- 15800312 TI - Improving accrual of older persons to cancer treatment trials: a randomized trial comparing an educational intervention with standard information: CALGB 360001. AB - PURPOSE: To design and test a geriatric educational intervention to improve accrual of cancer patients age 65 years and older to cooperative group-sponsored treatment trials. METHODS: Main member institutions of the Cancer and Leukemia Group B (CALGB) and its affiliates were randomly assigned to receive standard information (n = 73) or educational intervention (n = 53). Standard information included CALGB Web site access and periodic notification about existing trials. The geriatric educational intervention included standard information plus: (1) an educational seminar; (2) educational materials; (3) a list of available protocols for use on charts; (4) a monthly e-mail and mail reminders for 1 year; and (5) a case discussion seminar. The main outcome was percentage of accrual of older persons to phase II and III treatment protocols after study initiation compared with baseline. RESULTS: There were 3,032 patients entered onto trials in the baseline year, and 2,160 and 1,239 during the 2 years postintervention, respectively. Overall percentage of accrual of older patients was 37% at baseline, and 33% and 31% during the first and second years after intervention. There was no improvement in accrual in the intervention versus control arm: 36% v 32% in the first year and 31% v 31% in the second year. CONCLUSION: Accrual of older patients was not increased by this intervention. Reasons for lack of effect include low intervention intensity, high baseline accrual rates, and closure of several high-accruing protocols during the study. More intense and multifaceted approaches will be needed to change physician (and patient) behavior and to increase accrual of older persons to clinical trials. PMID- 15800313 TI - Primary CNS lymphoma of T-cell origin: a descriptive analysis from the international primary CNS lymphoma collaborative group. AB - PURPOSE: To describe the demographic and tumor related characteristics and outcomes for patients with primary T-cell CNS lymphoma (TPCNSL). PATIENTS AND METHODS: A retrospective series of patients with TPCNSL was compiled from twelve cancer centers in seven countries. RESULTS: We identified 45 patients with a median age of 60 years (range, 3 to 84 years). Twenty (44%) had Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1. Twenty-six (58%) had involvement of a cerebral hemisphere and sixteen (36%) had lesions of deeper sites in the brain. Serum lactate dehydrogenase was elevated in 7 (32%) of 22 patients, and CSF protein was elevated in 19 of 24 patients (79%) with available data. The median disease-specific survival (DSS) was 25 months (95% CI, 11 to 38 months). The 2- and 5-year DSS were 51% (95% CI, 35% to 66%) and 17% (95% CI, 6% to 34%), respectively. Univariate and multivariate analyses were conducted for age ( 60 years), PS (0 or 1 v 2, 3, or 4), involvement of deep structures of the CNS (no v yes), and methotrexate (MTX) use in the primary treatment (yes v no). Only PS and MTX use were significantly associated with better outcome with hazard ratios of 0.2 (95% CI, 0.1 to 0.4) and 0.4 (95% CI, 0.2 to 0.8), respectively. CONCLUSION: This is the largest series ever assembled of TPCNSL. The presentation and outcome appear similar to that of B cell PCNSL. PS 0 or 1 and administration of MTX are associated with better survival. PMID- 15800314 TI - Concurrent administration of high-dose rituximab before and after autologous stem cell transplantation for relapsed aggressive B-cell non-Hodgkin's lymphomas. AB - PURPOSE: We investigated the efficacy and safety of administering high-dose rituximab (HD-R) in combination with high-dose carmustine, cytarabine, etoposide, and melphalan chemotherapy and autologous stem-cell transplantation (SCT) in patients with recurrent B-cell aggressive non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Sixty-seven consecutive patients were treated. Rituximab was administered during stem-cell mobilization (1 day before chemotherapy at 375 mg/m(2) and 7 days after chemotherapy at 1,000 mg/m(2)), together with granulocyte colony-stimulating factor 10 mug/kg and granulocyte-macrophage colony stimulating factor 250 microg/m(2) administered subcutaneously daily. HD-R of 1,000 mg/m(2) was administered again days 1 and 8 after transplantation. The results of this treatment were retrospectively compared with those of a historical control group receiving the same preparative regimen without rituximab. RESULTS: With a median follow-up time for the study group of 20 months, the overall survival rate at 2-years was 80% (95% CI, 65% to 89%) for the study group and 53% (95% CI, 34% to 69%) for the control group (P = .002). Disease-free survival was 67% (95% CI, 51% to 79%) for the study group and 43% (95% CI, 26% to 60%) for the control group (P = .004). The median time to recovery of absolute neutrophil count to >/= 500 cells/microL was 11 days (range, 8 to 37 days) for the rituximab group and 10 days (range, 8 to 17 days) for the matched control group (P = .001). However, infections were not significantly increased in patients treated with rituximab. CONCLUSION: The results of this study suggest that using HD-R and autologous SCT is a feasible and promising treatment for patients with B-cell aggressive NHL. PMID- 15800315 TI - Prognostic significance of allelic imbalance at the c-kit gene locus and c-kit overexpression by immunohistochemistry in pediatric osteosarcomas. AB - PURPOSE: Since the recent development of biologic agents targeting oncogenes, increasing attention has been focused on determining the role of tyrosine kinase receptors in the pathogenesis of tumors. Our study was designed to investigate the status of region 4q12, which contains the candidate gene c-kit, and the expression of c-kit by immunohistochemistry (IHC). PATIENTS AND METHODS: Paired blood and biopsy specimens of 68 children treated for high-grade primary osteosarcomas were collected. Microsatellite analysis at two genomic sites containing c-kit gene was performed on paired DNA using a sensible fluorescent polymerase chain reaction technology. To confirm the DNA data, we studied c-kit protein expression by IHC in 56 available paraffin-embedded tumor tissues. RESULTS: The frequency of allelic imbalance (AI) at locus 4q12 was 39% in the overall population. In agreement with previous studies, we did not detect microsatellite instability, allowing us to hypothesize that this pathway is not implicated. Furthermore, the normal status at locus 4q12 was associated with a significantly better survival in the whole osteosarcoma population (P = .05). IHC overexpression of c-kit was concordant in all cases presenting an AI. However, normal status at locus 4q12 was correlated to an absence of c-kit protein expression in 19 (65.5%) of 29 informative cases. CONCLUSION: Allelotyping of locus 4q12, which contains the c-kit gene, could help pediatric osteosarcoma prognostic screening and showed a strong correlation with overexpression of c-kit protein. These results allowed us to hypothesize that, in some cases, a mutation of c-kit gene could lead to a protein overexpression. PMID- 15800316 TI - Serial evaluation of academic and behavioral outcome after treatment with cranial radiation in childhood. AB - PURPOSE: To evaluate academic and behavioral outcome in radiated survivors of posterior fossa (PF) tumors. PATIENTS AND METHODS: Fifty-three patients (36 males) treated for malignant PF tumors were seen for evaluation of academics and/or behavioral functioning. Forty-six patients were treated for medulloblastoma, and seven patients were treated for ependymoma. Fourteen patients were treated with reduced-dose cranial radiation, and 34 patients were treated with standard-dose cranial radiation (dose was not available for four patients). All patients received an additional boost to the PF. One patient was treated with PF radiation only. Standardized achievement tests and behavioral questionnaires were administered at different times after diagnosis for each child. First, the influence of demographic and medical variables on outcome was examined. Second, the rate of change in scores was determined using mixed model regression for patients seen for serial assessment. RESULTS: The presence of hydrocephalus was related to poorer academics, but outcome was not related to radiation dose, extent of surgery, or treatment with chemotherapy. Younger age predicted poor reading ability and lower parent rating of academic achievement. Children's performance declined for spelling, mathematics, and reading. Significant declines were also evident in parent and teacher's ratings of academic ability. Behavioral functioning was generally not related to medical and demographic variables, and few clinically significant problems in externalizing behavior were evident. Increases in social and attention problems emerged over time. CONCLUSION: Cranial radiation is associated with declines in academic ability, social skills, and attention. However, neither psychological distress nor behavior problems were a significant concern for this sample. PMID- 15800317 TI - Early responses to chemotherapy of normal and malignant hematologic cells are prognostic in children with acute lymphoblastic leukemia. AB - PURPOSE: Improved cure rates for children with acute lymphoblastic leukemia (ALL) have resulted from better relapse prediction, using clinical and laboratory features at diagnosis, and more intensive therapy in high-risk patients. More recently, measurements of the variation in the response of malignant lymphoblasts to chemotherapy in vivo have further improved relapse prediction. It is unknown whether the variation in the response of nonmalignant hematologic cells after chemotherapy correlates with the response of lymphoblasts or risk of relapse. PATIENTS AND METHODS: We retrospectively evaluated myelosuppression during induction and consolidation chemotherapy in 227 children uniformly treated for ALL on consecutive Australian and New Zealand Children's Cancer Study Group protocols. The early response to treatment was assessed in a representative subset (n = 62) by determining minimal residual disease (MRD) level by molecular techniques on the end-of-induction bone marrow sample. RESULTS: We found that a slow rate of myeloid recovery at the end of induction chemotherapy, reflected in a low absolute neutrophil count (ANC), was highly predictive of relapse (P < .0001). Additionally, patients with a high end-of-induction MRD level had a high risk of relapse (P = .001). Multivariate analysis confirmed the independent prognostic significance of MRD and ANC at the end of induction chemotherapy (P < .05). There was no significant association between other measures of myelotoxicity and MRD or relapse. CONCLUSION: We conclude that the responses of normal myeloid cells and malignant lymphoblasts to chemotherapy predict outcome by distinct mechanisms. While these results are promising, their use in the clinical setting needs to be examined in a future randomized controlled trial. PMID- 15800318 TI - Risk of new cancers after radiotherapy in long-term survivors of retinoblastoma: an extended follow-up. AB - PURPOSE: Many children diagnosed with retinoblastoma (Rb) survive into adulthood and are prone to subsequent cancers, particularly hereditary patients, who have germline Rb-1 mutations. We have extended the follow-up of a large cohort of Rb patients for 7 more years to provide new information on the risk of additional cancers after radiotherapy in long-term survivors. PATIENTS AND METHODS: We analyzed the risk of new cancers through 2000 in 1,601 Rb survivors, diagnosed from 1914 to 1984, at two US medical centers. The standardized incidence ratio (SIR) was calculated as the ratio of the observed number of cancers after hereditary and nonhereditary Rb to the expected number from the Connecticut Tumor Registry. The cumulative incidence of a new cancer after hereditary and nonhereditary Rb and radiotherapy was calculated with adjustment for competing risk of death. RESULTS: Subsequent cancer risk in 963 hereditary patients (SIR, 19; 95% CI, 16 to 21) exceeded the risk in 638 nonhereditary Rb patients (SIR, 1.2; 95% CI, 0.7 to 2.0). Radiation further increased the risk of another cancer in hereditary patients by 3.1-fold (95% CI, 2.0 to 5.3). Hereditary patients continued to be at significantly increased risk for sarcomas, melanoma, and cancers of the brain and nasal cavities. The cumulative incidence for developing a new cancer at 50 years after diagnosis of Rb was 36% (95% CI, 31% to 41%) for hereditary and 5.7% (95% CI, 2.4% to 11%) for nonhereditary patients. CONCLUSION: Hereditary Rb predisposes to a variety of new cancers over time, with radiotherapy further enhancing the risk of tumors arising in the radiation field. PMID- 15800319 TI - Unequivocal delineation of clinicogenetic subgroups and development of a new model for improved outcome prediction in neuroblastoma. AB - PURPOSE: Neuroblastoma is a genetically heterogeneous pediatric tumor with a remarkably variable clinical behavior ranging from widely disseminated disease to spontaneous regression. In this study, we aimed for comprehensive genetic subgroup discovery and assessment of independent prognostic markers based on genome-wide aberrations detected by comparative genomic hybridization (CGH). MATERIALS AND METHODS: Published CGH data from 231 primary untreated neuroblastomas were converted to a digitized format suitable for global data mining, subgroup discovery, and multivariate survival analyses. RESULTS: In contrast to previous reports, which included only a few genetic parameters, we present here for the first time a strategy that allows unbiased evaluation of all genetic imbalances detected by CGH. The presented approach firmly established the existence of three different clinicogenetic subgroups and indicated that chromosome 17 status and tumor stage were the only independent significant predictors for patient outcome. Important new findings were: (1) a normal chromosome 17 status as a delineator of a subgroup of presumed favorable-stage tumors with highly increased risk; (2) the recognition of a survivor signature conferring 100% 5-year survival for stage 1, 2, and 4S tumors presenting with whole chromosome 17 gain; and (3) the identification of 3p deletion as a hallmark of older age at diagnosis. CONCLUSION: We propose a new regression model for improved patient outcome prediction, incorporating tumor stage, chromosome 17, and amplification/deletion status. These findings may prove highly valuable with respect to more reliable risk assessment, evaluation of clinical results, and optimization of current treatment protocols. PMID- 15800320 TI - Merkel cell carcinoma: prognosis and treatment of patients from a single institution. AB - PURPOSE: Merkel cell carcinoma (MCC) is an uncommon cutaneous malignancy. Most reports consist of single-institution experiences of fewer than 30 patients. The natural history of MCC is poorly defined. PATIENTS AND METHODS: A review was performed of Memorial Sloan-Kettering Cancer Center's MCC database, identifying 251 patients who had been treated between 1970 and 2002. Patient, tumor, and treatment-related factors were analyzed for their association with recurrence and survival. RESULTS: The average follow-up for all patients was 40 months and 46 months for patients alive at last follow-up. The 5-year disease-specific survival rate was 64%. Disease stage was the only independent predictor of survival (stage I, 81%; stage II, 67%; stage III, 52%; stage IV, 11%; P = .001). Pathologic staging of the draining nodal basin was performed in 71 (40%) of 177 patients who presented with clinically negative nodes, and 16 of these patients (23%) were found to have node-positive disease. Pathologic nodal staging was associated with improved stage-specific survival probabilities (clinical node-negative, 75% v pathologic node-negative disease, 97%; P = .009) and decreased nodal recurrence (44% v 11%, P < .001). The median time to recurrence was 9 months, and 102 patients (43%) recurred. Local recurrence developed in 8% of patients after margin-negative excision. CONCLUSION: These data demonstrate that the natural history of MCC is variable and dependent on the stage of disease at presentation. Pathologic nodal staging identifies a group of patients with excellent long-term survival. After margin-negative excision and pathologic nodal staging, local and nodal recurrence rates are low. PMID- 15800321 TI - Chemoradiation with and without surgery in patients with locally advanced squamous cell carcinoma of the esophagus. AB - PURPOSE: Combined chemoradiotherapy with and without surgery are widely accepted alternatives for the curative treatment of patients with locally advanced esophageal cancer. The value of adding surgery to chemotherapy and radiotherapy is unknown. PATIENTS AND METHODS: Patients with locally advanced squamous cell carcinoma (SCC) of the esophagus were randomly allocated to either induction chemotherapy followed by chemoradiotherapy (40 Gy) followed by surgery (arm A), or the same induction chemotherapy followed by chemoradiotherapy (at least 65 Gy) without surgery (arm B). Primary outcome was overall survival time. RESULTS: The median observation time was 6 years. The analysis of 172 eligible, randomized patients (86 patients per arm) showed overall survival to be equivalent between the two treatment groups (log-rank test for equivalence, P < .05). Local progression-free survival was better in the surgery group (2-year progression free survival, 64.3%; 95% CI, 52.1% to 76.5%) than in the chemoradiotherapy group (2-year progression-free survival, 40.7%; 95% CI, 28.9% to 52.5%; hazard ratio [HR] for arm B v arm A, 2.1; 95% CI, 1.3 to 3.5; P = .003). Treatment-related mortality was significantly increased in the surgery group than in the chemoradiotherapy group (12.8% v 3.5%, respectively; P = .03). Cox regression analysis revealed clinical tumor response to induction chemotherapy to be the single independent prognostic factor for overall survival (HR, 0.30; 95% CI, 0.19 to 0.47; P < .0001). CONCLUSION: Adding surgery to chemoradiotherapy improves local tumor control but does not increase survival of patients with locally advanced esophageal SCC. Tumor response to induction chemotherapy identifies a favorable prognostic group within these high-risk patients, regardless of the treatment group. PMID- 15800322 TI - Low microsatellite instability is associated with poor prognosis in stage C colon cancer. AB - PURPOSE: The significance of low microsatellite instability (MSI-L) in colorectal cancer is poorly understood. No clear biologic distinction has been found between MSI-L and microsatellite stable (MSS) colorectal cancer, and these two phenotypes are usually combined when analyzed against the well-defined high MSI (MSI-H) phenotype. Evidence is emerging that an O(6)-methylguanine DNA methyltransferase (MGMT) gene defect is associated with MSI-L. Therefore, to further define this phenotype, we undertook a detailed analysis of the prognostic significance of MSI L and loss of MGMT expression in colon cancer. PATIENTS AND METHODS: The study cohort was 183 patients with clinicopathologic stage C colon cancer who had not received adjuvant therapy. We analyzed MSI status, MGMT, and mismatch repair protein expression, as well as MGMT and p16 promoter hypermethylation. RESULTS: We showed that MSI-L defines a group of patients with poorer survival (P = .026) than MSS patients, and that MSI-L was an independent prognostic indicator (P = .005) in stage C colon cancer. Loss of MGMT protein expression was associated with the MSI-L phenotype but was not a prognostic factor for overall survival in colon cancer. p16 methylation was significantly less frequent in MSI-L than in MSI-H and MSS tumors and was not associated with survival. CONCLUSION: MSI-L characterizes a distinct subgroup of stage C colon cancer patients, including the MSI-L subset of proximal colon cancer, who have a poorer outcome. Neither the MGMT defect nor p16 methylation are likely to contribute to the worse prognosis of the MSI-L phenotype. PMID- 15800323 TI - Outcome results of the 1996-1999 patterns of care survey of the national practice for patients receiving radiation therapy for carcinoma of the esophagus. AB - PURPOSE: A Patterns of Care Study of patients treated from 1996 to 1999 evaluated the national practice for patients receiving radiation therapy for carcinoma of the esophagus in the United States. METHODS: A national survey was conducted at 59 institutions in a stratified random sample selected from a master list of radiation therapy facilities throughout the United States. Patient, tumor, and treatment characteristics were evaluated. Multivariate comparisons of survival times were made using the Cox proportional hazards model. RESULTS: Adenocarcinoma was diagnosed in 51% of patients and squamous cell carcinoma in 49% of patients. Sixteen percent of patients were clinical stage (CS) I (using the 1983 American Joint Committee on Cancer system), 39% were CS II, and 33% were CS III. Significant variables in the multivariate analysis of survival times included clinical stage, treatment approach, and facility size. Patients with CS III disease had a higher hazard risk of death as compared with CS I patients (hazard ratio [HR], 2.01; P = .001), whereas those treated with chemoradiotherapy followed by surgery (HR, 0.32; P < .0001) had a decreased risk of death compared with chemoradiotherapy-only patients. Patients at small centers had a higher risk of death (HR, 1.32; P = .03) compared with patients treated at larger facilities. CONCLUSION: Concurrent chemoradiotherapy continued to be the most commonly utilized treatment approach during the time period studied. The observation that patients undergoing surgical resection following chemoradiation have a decreased HR or chance of death compared with other treatment schemes supports the need for a randomized trial comparing these strategies. PMID- 15800324 TI - Combining gemcitabine and capecitabine in patients with advanced biliary cancer: a phase II trial. AB - PURPOSE: Biliary cancer has a poor prognosis, and chemotherapy has had little impact. The objectives of this trial were to determine the response rate, time to disease progression, survival, and safety profile of the combination of gemcitabine and capecitabine (GemCap) in patients with advanced biliary cancer. PATIENTS AND METHODS: Eligible patients had pathologically proven, locally advanced or metastatic adenocarcinoma arising from the intra- and extrahepatic bile ducts or gallbladder with no prior chemotherapy. Patients were treated on a 3-week cycle consisting of capecitabine at 650 mg/m(2) orally twice a day for 14 days and gemcitabine at a fixed dose of 1,000 mg/m(2) intravenously over 30 minutes on days 1 and 8. RESULTS: Forty-five patients were enrolled between July 2001 and January 2004. Fifty-three percent of patients had cholangiocarcinoma, 47% had gallbladder cancer, and 89% had metastatic disease. The overall objective response rate was 31%, with an additional 42% of patients with stable disease, for a disease control rate of 73%. The median overall survival time was 14 months (95% CI, 7.3 months to not available), and the median progression-free survival time was 7 months (95% CI, 4.6 to 11.8 months). This chemotherapy combination was generally well tolerated. Transient neutropenia, thrombocytopenia, fatigue, and hand-foot syndrome were commonly observed but were easily managed without discontinuing further treatment. CONCLUSION: The significant antitumor activity combined with a mild toxicity profile seen in this study argue that GemCap chemotherapy may benefit patients with advanced biliary cancer. This regimen warrants further evaluation in a randomized study with survival and quality of life end points. PMID- 15800325 TI - Prognostic role of interleukin-1beta gene and interleukin-1 receptor antagonist gene polymorphisms in patients with advanced gastric cancer. AB - PURPOSE: A high interleukin-1beta (IL-1B) and interleukin-1 receptor antagonist (IL-RN) ratio underlies an unfavorable proinflammatory status. Also, it seems to be involved in the mechanisms of cancer cachexia and tumor angiogenesis and metastasis. Two single nucleotide polymorphisms in IL-1B gene (IL-1B-511C/T,IL-1B 31T/C) and a variable number of tandem repeat polymorphisms in IL-RN gene (IL 1RNlong/2) enhance the circulating levels of the two cytokines. The prognostic role of IL-1B/IL-1RN genotypes was investigated in patients with relapsed and metastatic gastric cancer treated with palliative chemotherapy. PATIENTS AND METHODS: Before starting palliative chemotherapy, 123 prospectively enrolled patients supplied peripheral-blood samples for DNA extraction. Survival data were analyzed according to IL-1RN/IL-1B genotypes. RESULTS: Forty-two patients showed wild-type genotypes (IL-1RNlong/long, IL-1B-511C/C, and IL-1B-31T/T; group A). Forty-five patients showed the IL-1RN2 polymorphism, with wild-type IL-1B genotypes in seven patients and with IL-1B-511C/T and/or IL-1B-31T/C polymorphisms in 38 patients (group B). The remaining 36 patients demonstrated wild-type IL-1RN, with IL-1B-511C/T and/or IL-1B-31T/C polymorphisms (group C). In group A and B patients, the median progression-free survival (PFS) was 25 and 26 weeks, respectively, and median overall survival (OS) was 42 and 43 weeks, respectively. Group C patients showed worse PFS (median, 16 weeks) and OS (median, 28 weeks) than group A (P = .006 for PFS; P = .0001 for OS) and group B patients (P = .01 for PFS; P = .0001 for OS). The long/T/C haplotype was overrepresented in patients with shortened PFS (P = .001) and OS (P = .0005). CONCLUSION: In patients with advanced gastric cancer, IL-1B polymorphisms showed adverse prognostic influence when coupled with wild-type IL-1RN genotype. These findings deserve further investigation for potential anticancer activity of recombinant IL-RN. PMID- 15800327 TI - Extracapsular extension of pelvic lymph node metastases from urothelial carcinoma of the bladder is an independent prognostic factor. AB - PURPOSE: To analyze the prognostic impact of risk factors for urothelial carcinoma of the bladder (UCB) with pelvic lymph node (LN) metastases. PATIENTS AND METHODS: We analyzed a consecutive series of 507 patients with UCB who were preoperatively staged N0M0. One hundred one of 124 eligible patients who were treated with radical cystectomy and standardized extended bilateral pelvic lymphadenectomy with curative intent and had postoperatively confirmed LN metastases were evaluated in regard to recurrence-free and overall survival. RESULTS: A median of 22 nodes per patient (range, 10 to 43 nodes) were removed and examined. Median recurrence-free and overall survival durations were 17 months and 21 months (range for both, 1 to 191 months), respectively. In the multivariate analysis for recurrence-free survival, extracapsular extension of LN metastases was the strongest prognostic factor (P = .019). Other variables such as tumor stage (pT1/2 v pT3 and pT4), the number (< five v >/= five), and the percentage (< 20% v >/= 20%) of metastatic nodes had a significant influence on recurrence-free and overall survival in the univariate analysis. However, they all failed to be significant prognostic factors in the multivariate analysis. CONCLUSION: The results of this study indicate that radical cystectomy with bilateral lymphadenectomy can have a curative effect in a subset of patients with pelvic LN positive UCB. Provided that a representative number of LNs are removed by meticulous lymphadenectomy and that a thorough histologic examination is performed, prognosis for such patients can be determined quite reliably. In the multivariate analysis, extracapsular extension of LN metastases was the strongest factor predicting prognosis. PMID- 15800326 TI - Adoptive cell transfer therapy following non-myeloablative but lymphodepleting chemotherapy for the treatment of patients with refractory metastatic melanoma. AB - PURPOSE: We investigated the combination of lymphodepleting chemotherapy followed by the adoptive transfer of autologous tumor reactive lymphocytes for the treatment of patients with refractory metastatic melanoma. PATIENTS AND METHODS: Thirty-five patients with metastatic melanoma, all but one with disease refractory to treatment with high-dose interleukin (IL) -2 and many with progressive disease after chemotherapy, underwent lymphodepleting conditioning with two days of cyclophosphamide (60 mg/kg) followed by five days of fludarabine (25 mg/m(2)). On the day following the final dose of fludarabine, all patients received cell infusion with autologous tumor-reactive, rapidly expanded tumor infiltrating lymphocyte cultures and high-dose IL-2 therapy. RESULTS: Eighteen (51%) of 35 treated patients experienced objective clinical responses including three ongoing complete responses and 15 partial responses with a mean duration of 11.5 +/- 2.2 months. Sites of regression included metastases to lung, liver, lymph nodes, brain, and cutaneous and subcutaneous tissues. Toxicities of treatment included the expected hematologic toxicities of chemotherapy including neutropenia, thrombocytopenia, and lymphopenia, the transient toxicities of high dose IL-2 therapy, two patients who developed Pneumocystis pneumonia and one patient who developed an Epstein-Barr virus-related lymphoproliferation. CONCLUSION: Lymphodepleting chemotherapy followed by the transfer of highly avid antitumor lymphocytes can mediate significant tumor regression in heavily pretreated patients with IL-2 refractory metastatic melanoma. PMID- 15800328 TI - Effects of parenteral hydration in terminally ill cancer patients: a preliminary study. AB - PURPOSE: Most patients with cancer develop decreased oral intake and dehydration before death. This study aimed to determine the effect of parenteral hydration on overall symptom control in terminally ill cancer patients with dehydration. PATIENTS AND METHODS: Patients with clinical evidence of mild to moderate dehydration and a liquid oral intake less than 1,000 mL/day were randomly assigned to receive either parenteral hydration with 1,000 mL (treatment group) or placebo with 100 mL normal saline administered over 4 hours for 2 days. Patients were evaluated for target symptoms (hallucinations, myoclonus, fatigue, and sedation), global well-being, and overall benefit. RESULTS: Twenty-seven patients randomly assigned to the treatment group had improvement in 53 (73%) of their 73 target symptoms versus 33 (49%) of 67 target symptoms in the placebo group (n=22; P = .005). Fifteen (83%) of 18 and 15 (83%) of 18 patients had improved myoclonus and sedation after hydration versus eight (47%) of 17 and five (33%) of 15 patients after placebo (P = .035 and P = .005, respectively). There were no significant differences of improvement in hallucinations or fatigue between groups. When blinded to treatment, patients (17 [63%] of 77) and investigators (20 [74%] of 27) perceived hydration as effective compared with placebo in nine (41%) of 22 patients (P = .78) and 12 (54%) of 22 investigators (P = .15), respectively. The intensity of pain and swelling at the injection site were not significantly different between groups. CONCLUSION: Parenteral hydration decreased symptoms of dehydration in terminally ill cancer patients who had decreased fluid intake. Hydration was well tolerated, and a placebo effect was observed. Studies with larger samples and a longer follow-up period are justified. PMID- 15800329 TI - Randomized phase II study of temozolomide and radiotherapy compared with radiotherapy alone in newly diagnosed glioblastoma multiforme. AB - PURPOSE: Surgery remains the standard treatment for glioma, followed by radiotherapy (RT) with or without chemotherapy. Despite multidisciplinary approaches, the median survival time for patients with glioblastoma multiform (GBM) remains at less than 1 year from initial diagnosis. Temozolomide (TMZ), an oral alkylating agent, has shown promising activity in the treatment of malignant gliomas. We conducted a multicenter randomized phase II study comparing the efficacy and safety of TMZ administered concomitantly and sequentially to RT versus RT alone in patients with newly diagnosed GBM. PATIENTS AND METHODS: One hundred thirty patients with pathologically confirmed, newly diagnosed GBM were randomly assigned (110 assessable patients) to receive either TMZ 75 mg/m(2)/d orally, concomitantly with RT (60 Gy in 30 fractions; group A, n = 57), followed by six cycles of TMZ (150 mg/m(2) on days 1 through 5 and 15 to 19 every 28 days), or RT alone (60 Gy in 30 fractions; group B, n = 53). RESULTS: Median time to progression was 10.8 months in group A and 5.2 months in group B (P = .0001). One-year progression-free survival rate was 36.6% in group A and 7.7% in group B. Median overall survival (OS) time was also significantly better in group A versus group B (13.4 v 7.7 months, respectively; P < .0001), as was the 1-year OS at 56.3% v 15.7% (P < .0001), respectively. Toxicity was mainly hematologic. One patient with grade 4 myelotoxicity died as a result of sepsis. The other side effects were mild. CONCLUSION: TMZ combined with RT (concomitantly and sequentially) seems to be more effective than RT alone in patients with newly diagnosed GBM. The combined-modality treatment was well tolerated. PMID- 15800330 TI - Effectiveness of physical activity on cardiorespiratory fitness and health related quality of life in young and middle-aged cancer patients shortly after chemotherapy. AB - PURPOSE: To evaluate the effectiveness of a supervised home-based flexible training program on cardiorespiratory fitness (CRF), mental distress, and health related quality of life (HRQOL) parameters in young and middle-aged cancer patients shortly after curative chemotherapy. PATIENTS AND METHODS: One hundred eleven patients age 18 to 50 years who had received chemotherapy for lymphomas or breast, gynecologic, or testicular cancer completed the trial. These patients were randomly allocated to either an intervention group (n = 59), which underwent a 14-week training program, or a control group (n = 52) that received standard care. Primary outcome was change in CRF, as determined by Astrand-Rhyming indirect bicycle ergometer test (maximum oxygen uptake [VO(2max)]), between baseline (T0) and follow-up (T1). Secondary outcomes were mental distress, as assessed by the Hospital Anxiety and Depression Scale, and HRQOL, as assessed by the European Organisation for Research and Treatment of Cancer Core Quality of Life Questionnaire. Two-way analysis of covariance was used to analyze changes from T0 to T1. RESULTS: VO(2max) increased by 6.4 mL/kg(-1)/min(-1) in patients in the intervention group and by 3.1 mL/kg(-1)/min(-1) in patients in the control group (P < .01). The fatigue score decreased by 17.0 points in the control group compared with only 5.8 points in the intervention group (P < .01). There were no intergroup differences in mental distress or HRQOL. CONCLUSION: A supervised, home-based, flexible training program has significant effect on CRF in young and middle-aged cancer patients shortly after curative chemotherapy, but it has no favorable effect on patients' experience of fatigue, mental distress, or HRQOL. PMID- 15800331 TI - Study of anxiety disorder and depression in long-term survivors of testicular cancer. AB - PURPOSE: To increase our knowledge of the prevalence of anxiety disorder and depression in long-term testicular cancer survivors (TCSs), and to identify variables associated with such caseness. PATIENTS AND METHODS: Participants were 1,408 TCSs treated between 1980 and 1994 in Norway. Participants provided information about their medical, social, and familial situation on a questionnaire. They also completed the Hospital Anxiety and Depression Scale (HADS). Anxiety disorder and depression were defined by a score >/= 8 on the HADS subscales. The prevalence rates were compared with age-adjusted norm data. RESULTS: HADS-defined anxiety disorder was more prevalent in TCSs (19.2%; 95% CI, 17.2% to 21.3%) than in the norm sample (13.5%; 95% CI, 13.1% to 13.9%; P < .001), whereas the prevalence of HADS-defined depression did not differ from the norm (TCSs, 9.7%; 95% CI, 8.1% to 11.2% v norm, 10.1%, 95% CI, 9.5 to 10.5; P = .56). The relative risk for anxiety disorder was 1.49 (95% CI, 1.31 to 1.69) and for depression the relative risk was 0.96 (95% CI, 0.81 to 1.14) in TCSs compared with norm. In multivariate analyses, HADS-defined anxiety disorder in TCSs was associated with young age, peripheral neuropathy, economic problems, alcohol problems, sexual problems, relapse anxiety, and having been treated for mental problems. CONCLUSION: Long-term TCSs have an increased risk of HADS-defined anxiety disorder that warrants clinical attention. Checking easily available demographic and TC-related data and use of a simple screening test such as HADS assists the identification of TCSs with anxiety disorder. PMID- 15800332 TI - Arsenic trioxide: new clinical experience with an old medication in hematologic malignancies. AB - Arsenic trioxide has shown great promise in the treatment of patients with relapsed or refractory acute promyelocytic leukemia (APL). In clinical trials, arsenic trioxide induces complete remission in 87% of patients and molecular remission in 83% of patients. Two-year overall and relapse-free survival estimates are 63% and 49%, respectively. Treatment with arsenic trioxide may be associated with the APL differentiation syndrome, leukocytosis, and electrocardiographic abnormalities. The expanded use of arsenic trioxide in APL for postremission therapy, in conjunction with transplantation, and in patients with newly diagnosed APL is under investigation. The multiple mechanisms of action of arsenic trioxide suggest that it may have antitumor activity in malignancies other than APL and that it may be used in combination with other agents to expand its potential use. This article reviews the clinical use of arsenic trioxide to date and discusses new therapeutic strategies evolving from its diverse biologic activities. PMID- 15800333 TI - Possible future issues in the treatment of glioblastomas: special emphasis on cell migration and the resistance of migrating glioblastoma cells to apoptosis. AB - PURPOSE: The present review aims to emphasize that malignant gliomas are characterized by the diffuse invasion of distant brain tissue by a myriad of single migrating cells that exhibit decreased levels of apoptosis (programmed cell death type I), thus a resistance to cytotoxic insult. METHODS: The present review surveys the molecular mechanisms of migration in malignant gliomas and potential issues arising from treatments, in addition to relationships between glioma cell migration and resistance to apoptosis in terms of the molecular signaling pathways. RESULTS: Clinical and experimental data demonstrate that glioma cell migration is a complex combination of multiple molecular processes, including the alteration of tumor cell adhesion to a modified extracellular matrix, the secretion of proteases by the cells, and modifications to the actin cytoskeleton. Intracellular signaling pathways involved in the acquisition of resistance to apoptosis by migrating glioma cells concern PI3K, Akt, mTOR, NF kappaB, and autophagy (programmed cell death type II). CONCLUSION: A number of signaling pathways can be constitutively activated in migrating glioma cells, thus rendering these cells resistant to cytotoxic insults. However, these pathways are not all constitutively activated at the same time in any one glioma. Particular inhibitors should therefore only be chosen if the target is present in the tumor tissue, but this is only possible if individual patients are submitted to the molecular profiling of their tumors before undergoing any treatment to combat their migratory glioma cells. Specific antimigratory compounds should be added to conventional radio- and/or chemotherapy. PMID- 15800334 TI - Side effects and good effects from new chemotherapeutic agents. Case 1. Gefitinib induced interstitial fibrosis. PMID- 15800335 TI - Side effects and good effects from new chemotherapeutic agents. Case 2. Thalidomide-induced interstitial pneumonitis. PMID- 15800336 TI - Side effects and good effects from new chemotherapeutic agents. Case 3. Bortezomib in primary refractory plasmacytoma. PMID- 15800337 TI - Witness. PMID- 15800339 TI - Adjuvant interferon therapy for melanoma. PMID- 15800341 TI - Highly active antiretroviral therapy in AIDS-associated Kaposi's sarcoma (KS): implications for the design of therapeutic trials in patients with advanced symptomatic KS. PMID- 15800342 TI - Elevated body mass index predicts for longer overall survival duration in men with metastatic hormone-refractory prostate cancer. PMID- 15800343 TI - CA125 Nadir values as a prognostic factor in epithelial ovarian cancer. PMID- 15800346 TI - Rationing and decision making. PMID- 15800348 TI - Overestimation of lung cancer mortality in a computed tomography-screened population. PMID- 15800351 TI - Pattern of care at the end of life: does age make a difference? PMID- 15800352 TI - Looking for the role of NSAIDs in cancer pain. PMID- 15800354 TI - The paradox of promoting choice in a collectivist system. PMID- 15800355 TI - An antidote to the emerging two tier organ donation policy in Canada: the Public Cadaveric Organ Donation Program. AB - In Canada, as in many other countries, there exists an organ procurement/donation crisis. This paper reviews some of the most common kidney procurement and allocation programmes, analyses them in terms of public and private administration, and argues that privately administered living donor models are an inequitable stopgap measure, the good intentions of which are misplaced and opportunistic. Focusing on how to improve the publicly administered equitable cadaveric donation programme, and at the same time offering one possible explanation for its current failure, it is suggested that the simple moral principle of "give and you shall receive", already considered by some, be extended further. This would allow for those who are willing to sign up to be a public cadaveric donor be given a priority for receiving an organ donation should they ever require it. It is argued that this priority may provide the motivation to give that is so far lacking in Canada. This model is called the Public Cadaveric Organ Donation Program. PMID- 15800356 TI - How should we measure informed choice? The case of cancer screening. AB - Informed choice is increasingly recognised as important in supporting patient autonomy and ensuring that people are neither deceived nor coerced. In cancer screening the emphasis has shifted away from just promoting the benefits of screening to providing comprehensive information to enable people to make an informed choice. Cancer screening programmes in the UK now have policies in place which state that it is their responsibility to ensure that individuals are making an individual informed choice. There is a need to evaluate whether such policies mean that those people invited for screening are making informed choices, and how comprehensive information affects other variables such as uptake, cost effectiveness, and satisfaction. At the present time, there is no validated measure of informed choice in cancer screening. Such a measure could be used to evaluate the effectiveness of interventions to increase informed choice and levels of informed choice in a population invited for screening. It could encourage health professionals to be accountable. Factors important when measuring informed choice in cancer screening include an individual's understanding of the limitations of screening, the ability to make an autonomous choice, and the difference between choice and behaviour. PMID- 15800357 TI - Laboratory animals and the art of empathy. AB - Consistency is the hallmark of a coherent ethical philosophy. When considering the morality of particular behaviour, one should look to identify comparable situations and test one's approach to the former against one's approach to the latter. The obvious comparator for animal experiments is non-consensual experiments on people. In both cases, suffering and perhaps death is knowingly caused to the victim, the intended beneficiary is someone else, and the victim does not consent. Animals suffer just as people do. As we condemn non-consensual experiments on people, we should, if we are to be consistent, condemn non consensual experiments on animals. The alleged differences between the two practices often put forward do not stand up to scrutiny. The best guide to ethical behaviour is empathy--putting oneself in the potential victim's shoes. Again to be consistent, we should empathise with all who may be adversely affected by our behaviour. By this yardstick, too, animal experiments fail the ethical test. PMID- 15800358 TI - Pain, vivisection, and the value of life. PMID- 15800360 TI - Legitimate requests and indecent proposals: matters of justice in the ethical assessment of phase I trials involving competent patients. AB - The death of Jesse Gelsinger in 1999 during a gene therapy trial raised many questions about the ethical review of medical research. Here, the author argues that the principle of justice is interpreted too narrowly and receives insufficient emphasis and that what we permit in terms of bodily invasion affects the value we place on individuals. Medical research is a societally supported activity. As such, the author contends that justice requires that invasive medical research demonstrates sufficiently compelling societal benefit. Many consider this societal benefit to be self evident. However, medical research is a complex activity; it yields new treatments but also creates financial rewards and affects health resource allocation. As research evolves into a multibillion pound, multinational enterprise, justice requires a much broader analysis of societal benefit. Without such evaluation we risk undermining the value of bodily integrity and of research participants. PMID- 15800361 TI - Therapeutic optimism in the consent forms of phase 1 gene transfer trials: an empirical analysis. AB - BACKGROUND: "Therapeutic misconception" arises when human subjects interpret a clinical trial as aimed primarily at therapy rather than producing knowledge. Therapeutic misconceptions may be more prevalent in trials enrolling gravely ill subjects or involving novel and well publicized investigational agents. OBJECTIVE: To examine the extent to which investigators express therapeutic optimism in phase 1 human gene transfer consent documents, whether highly active gene transfer researchers are more prone to expressing therapeutic optimism, and whether consent forms have grown more optimistic in their descriptions of personal benefit over the last decade. DESIGN: Content analysis was performed on 277 consent documents to measure the number of sentences describing possibility of benefit, terminology used for experimental agents, the proportion of statements describing personal versus societal benefits, and whether investigators attempted to thwart therapeutic misconceptions. RESULTS: Consent forms generally used therapeutic terminology to describe study agents, devoted more sentences to describing possible personal benefits than to describing benefits to society, and infrequently explained that a particular benefit was unlikely. Consent documents used by highly active gene transfer researchers tended to portray significantly greater optimism about personal benefit than less active investigators, though they were also significantly more cautious with agent terminology. Finally, therapeutic optimism expressed in consent forms has declined over the past decade. CONCLUSIONS: Consent documents used in phase 1 gene transfer trials, although increasingly attentive to possible therapeutic misconceptions, are inappropriately optimistic about direct benefits of trial participation. Such optimism is expressed more emphatically in trials involving highly active gene transfer researchers as principal investigators. PMID- 15800362 TI - In the world of Dolly, when does a human embryo acquire respect? AB - For most of the 20th century, it was possible to regard fertilisation as the identifiable point when life begins, because this moment could be defined unequivocally and was thought to be the single most essential biological step in the establishment of a new human entity. Since the successful reproductive cloning of Dolly and other mammals, it is clear that any human cell has the potential to supply the full genome of an embryo, and hence a person, without going through fertilisation. At what point in time do such embryos acquire the respect accorded to human beings? The authors argue that the time of implantation is the most useful point at which the potential and the intention to create a new person are translated into reality, because from that point a new life develops. Implantation differentiates a somatic cell in culture (which is not due respect) from a human entity that has acquired its own identity and developmental potential. The authors examine the value of quickening or viability as alternative developmental stages in the process of acquiring respect for the Dolly embryo. PMID- 15800363 TI - The moral status of the embryo post-Dolly. AB - Cameron and Williamson have provided a provocative and timely review of the ethical questions prompted by the birth of Dolly. The question Cameron and Williamson seek to address is "In the world of Dolly, when does a human embryo acquire respect?". Their initial discussion sets the scene by providing a valuable overview of attitudes towards the embryo, summarising various religious, scientific, and philosophical viewpoints. They then ask, "What has Dolly changed?" and identify five changes, the first being that fertilisation is no longer required to create an embryo. Following this analysis they then ask when an embryo created other than by fertilisation begins to acquire respect. This paper explores the ethical and legal issues highlighted by Cameron and Williamson's paper. PMID- 15800364 TI - Challenges in the provision of ICU services to HIV infected children in resource poor settings: a South African case study. AB - The HIV/AIDS epidemic has placed increasing demands on limited paediatric intensive care services in developing countries. The decision to admit HIV infected children with Pneumocystis carinii pneumonia (PCP) into the paediatric intensive care unit (PICU) has to be made on the best available evidence of outcome and the ethical principles guiding appropriate use of scarce resources. The difficulty in confirming the diagnosis of HIV infection and PCP in infancy, issues around HIV counselling, and the variance in the outcome of HIV infected children with PCP admitted to the PICU in African studies compound this process. Pragmatic decision making will require evaluation of at least three ethical questions: are there clinical and moral reasons for admitting HIV positive children with PCP to the PICU, should more resources be committed to caring for HIV children who require the PICU, and how can we morally choose candidates for the PICU? Those working in the PICU in HIV endemic regions need to make difficult personal decisions on effective triage of admissions of HIV infected children with PCP based on individual case presentation, availability of resources, and applicable ethical principles. PMID- 15800365 TI - Just diagnosis? Preimplantation genetic diagnosis and injustices to disabled people. PMID- 15800366 TI - The case for physician assisted suicide: not (yet) proven. AB - The legalization of physician assisted suicide (PAS) in Oregon and physician assisted death (PAD) in The Netherlands has revitalized the debate over whether and under what conditions individuals should be able to determine the time and manner of their deaths, and whether they should be able to enlist the help of physicians in doing so. Although the change in the law is both dramatic and recent, the basic arguments for and against have not really changed since the issue was debated by Glanville Williams and Yale Kamisar nearly 50 years ago. In this paper, the author argues in favour of Kamisar's consequentialist framework. Any change in law and social policy should not be based solely on individual cases, heart wrenching though these may be. Instead, we need to assess the need for PAS, and weigh this against the risks of mistake and abuse. PMID- 15800367 TI - Scientific research is a moral duty. PMID- 15800368 TI - Barriers to the socialization of information. PMID- 15800369 TI - Mining for associations between text and brain activation in a functional neuroimaging database. AB - We describe a method for mining a neuroimaging database for associations between text and brain locations. The objective is to discover association rules between words indicative of cognitive function as described in abstracts of neuroscience papers and sets of reported stereotactic Talairach coordinates. We invoke a simple probabilistic framework in which kernel density estimates are used to model distributions of brain activation foci conditioned on words in a given abstract. The principal associations are found in the joint probability density between words and voxels. We show that the statistically motivated associations are well aligned with general neuroscientific knowledge. PMID- 15800370 TI - A probabilistic functional atlas of the human subthalamic nucleus. AB - This paper introduces a method for generation and validation of a probabilistic functional brain atlas of subcortical structures from electrophysiological and neuroimaging data. The method contains three major steps: (1) acquisition of pre, intra, and postoperative multimodal data; (2) selection of an accurate data set for atlas generation; and (3) generation of the atlas from the selected data set. The method is applied to construct the probabilistic functional atlas of the human subthalamic nucleus (STN). The STN atlas has been built from data collected during surgical treatment of 184 patients with Parkinson's disease. It is based on preoperative X-ray ventriculography imaging, intraoperative electrophysiological measurements and X-ray imaging, and postoperative neurological assessment. The atlas features a high resolution of 0.25 mm(3) and a high accuracy of 0.25 mm. It is dynamic and can be rapidly recalculated for arbitrary resolution and extended by adding new patient data. The atlas can easily be reformatted and warped to match patient-specific data. Its applications include planning of subthalamic stimulations and neuroscience research to study functional properties of the STN. The presented method is general and can be applied for constructing human and animal probabilistic brain atlases. PMID- 15800371 TI - Artificial neural networks and artificial organisms can predict Alzheimer pathology in individual patients only on the basis of cognitive and functional status. AB - Data from several studies have pointed out the existence of a strong correlation between Alzheimer's disease (AD) neuropathology and cognitive state. However, because of their highly complex and nonlinear relationship, it has been difficult to develop a predictive model for individual patient classification through traditional statistical approaches. When exposed to complex data sets, artificial neural networks (ANNs) can recognize patterns, learn the relationship of different variables, and address classification tasks. To predict the results of postmortem brain examinations, we applied ANNs to the Nun Study data set, a longitudinal epidemiological study, which includes annual cognitive and functional evaluation. One hundred seventeen subjects from the study participated in this analysis. We determined how demographic data and the cognitive and functional variables of each subject during the last year of her life could predict the presence of brain pathology expressed as Braak stages, neurofibrillary tangles (NFTs) and neuritic plaques (NPs) count in the neocortex and hippocampus, and brain atrophy. The result of this analysis was then compared with traditional statistical models. ANNs proved to be better predictors than Linear Discriminant Analysis in all experimentations (+ approximately 10% in overall accuracy), especially when assembled in Artificial Organisms (+ approximately 20% in overall accuracy). Demographic, cognitive, and clinical variables were better predictors of tangles count in the neocortex and in the hippocampus when compared to NPs count. These findings strengthen the hypothesis that neurofibrillary pathology may represent the major anatomic substrate of the cognitive impairment found in AD. PMID- 15800372 TI - An FPGA-based approach to high-speed simulation of conductance-based neuron models. AB - The constant requirement for greater performance in neural model simulation has created the need for high-speed simulation platforms. We present a generalized, scalable field programmable gate array (FPGA)-based architecture for fast computation of neural models and focus on the steps involved in implementing a single-compartment and a two-compartment neuron model. Based on timing tests, it is shown that FPGAs can outperform traditional desktop computers in simulating these fairly simple models and would most likely provide even larger performance gains over computers in simulating more complex models. The potential of this method for improving neural modeling and dynamic clamping is discussed. In particular, it is believed that this approach could greatly speed up simulations of both highly complex single neuron models and networks of neurons. Additionally, our design is particularly well suited to automated parameter searches for tuning model behavior and to real-time simulation. PMID- 15800373 TI - Insulysin: an allosteric enzyme as a target for Alzheimer's disease. AB - That the zinc metalloendopeptidase insulysin (insulin-degrading enzyme IDE) is a major b-amyloid (A(beta)) peptide-degrading enzyme in vivo is shown by the higher A(beta) peptide levels in the brain of an insulysin-deficient mouse. Insulysin was shown to initially cleave A(beta)1-40and A(beta)1-42 at His13-Gln14, His14 Gln15, and Phe19-Phe20. The insulysin-dependent cleavage of A(beta) prevents both the neurotoxic effects of the peptide as well as the ability of A(beta) to deposit onto synthetic amyloid plaques. The kinetics of the reaction of insulysin with the synthetic peptide substrate Abz-G-G-F-L-R-K-H-G-Q-EDDnp displays allosteric properties indicative of a regulated enzyme. Small peptide substrates increase the activity of insulysin toward the hydrolysis of A(beta)1-40 without affecting the activity of the enzyme toward insulin. These studies indicate that insulysin is a target for drug development in which small-molecule peptide analogs can be used to increase the rate of A(beta) clearance without affecting insulin levels. PMID- 15800374 TI - Prolonged morphine treatment selectively increases membrane recruitment of delta opioid receptors in mouse basal ganglia. AB - In recent years, we demonstrated that prolonged (48-h) treatment of rats or mice with selective m-opioid receptor ((mu)OR) agonists induced a translocation of delta-opioid receptors ((delta)ORs) from intracellular compartments to neuronal plasma membranes in the dorsal horn of the spinal cord. It remained to be determined whether this phenomenon also occurred in the brain. To resolve this issue, we analyzed by immunogold histochemistry the subcellular distribution of (delta)ORs in the nucleus accumbens, dorsal neostriatum, and frontal cortex in mice treated or not with morphine (48 h). We observed that prolonged treatment with morphine induced a translocation of (delta)ORs from intracellular to subplasmalemmal and membrane compartments in dendrites from both the nucleus accumbens and the dorsal neostriatum but not from the frontal cortex. We propose that this (mu)OR-(delta)OR interaction might prolong and modulate the sensitivity of neurons to opiates in specific target regions. PMID- 15800376 TI - Activity-dependent neurotrophic factor-9 and NAP promote neurite outgrowth in rat hippocampal and cortical cultures. AB - Activity-dependent neurotrophic factor (ADNF) is a novel, femtomolar-acting, glial-derived polypeptide (14 kDa) known to protect neurons from a variety of toxic insults. The active site for ADNF function is localized to a 9-amino-acid stretch (SALLRSIPA; ADNF-9). A few years later, a novel ADNF-9-like active peptide (NAPVSIPQ or NAP) was identified and shown to be expressed in the CNS and exhibit an activity profile similar to ADNF-9. Such studies suggest that ADNF-9 and NAP might function like other known neurotrophins and play a role in neural development and maintenance. The purpose of the present studies was to determine if ADNF-9 or NAP affects neurite outgrowth and synaptogenesis in rat hippocampal and cortical cultures. Using MAP2-FITC immunofluorescent labeling, we found that ADNF-9 and NAP promoted neurite outgrowth in a concentration-dependent manner, with maximal activity observed at femtomolar concentrations. Both peptides stimulated robust outgrowth in hippocampal cells (approximately 150% of control; p < 0.01) with a modest effect on cortical cells (approximately 20% of control; p < 0.05) similar to other known growth factors. However, the outgrowth-promoting effect was abolished in the absence of serum, suggesting that soluble factors might be necessary for the neurotrophic activity. Finally, we found that ADNF-9 and NAP increased synaptophysin expression in both rat hippocampal and cortical cultures. These results suggest that ADNF-9 and NAP might contribute to neuronal plasticity associated with development and repair after injury. PMID- 15800375 TI - Receptors for VIP and PACAP in guinea pig cerebral cortex: effects on cyclic AMP synthesis and characterization by 125I-VIP binding. AB - Receptors for vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating polypeptide (PACAP) in guinea pig cerebral cortex were characterized by (1) radioreceptor binding of 125I-labeled VIP (human/rat/porcine), and (2) cyclic AMP (cAMP) formation. Saturation analysis of 125I-VIP binding to membranes of guinea pig cerebral cortex resulted in a linear Scatchard plot, suggesting the presence of a single class of high-affinity receptor-binding sites, with a Kd of 0.63 nM and a B(max) of 77 fmol/mg protein. Various peptides from the PACAP/VIP/secretin family displaced the specific binding of 125I-VIP to guinea pig cerebrum with the relative rank order of potency: chicken VIP (cVIP) > or = PACAP38 approximately PACAP27 approximately guinea pig VIP (gpVIP) > or = mammalian (human/rat/porcine) VIP (mVIP) > peptide histidine-methionine (PHM) > peptide histidine-isoleucine (PHI) > secretin. Analysis of the competition curves revealed displacement of 125I-VIP from high- and lower-affinity binding sites, with IC50 values in the picomolar and the nanomolar range, respectively. About 70% of the specific 125I-VIP-binding sites in guinea pig cerebral cortex were sensitive to Gpp(NH)p, a nonhydrolyzable analog of GTP. Pituitary adenylate cyclase-activating polypeptide 38 (PACAP38), PACAP27, cVIP, gpVIP, mVIP, PHM, and PHI stimulated cAMP production in [3H]adenine-prelabeled slices of guinea pig cerebral cortex in a concentration-dependent manner. Of the tested peptides, the most effective were PACAP38 and PACAP27, which at a 1 microM concentration produced a 17- to 19-fold rise in cAMP synthesis, increasing the nucleotide production to approx 11% conversion above the control value. The three forms of VIP (cVIP, mVIP, and gpVIP) at the highest concentration used, i.e., 3 microM, produced net increases in cAMP production in the range of 8-9% conversion, whereas 5 microM PHM and PHI, by, respectively, 6.7% and 4.9% conversion. It is concluded that cerebral cortex of guinea pig contains VPAC- type receptors positively linked to cAMP formation. In addition, the observed stronger action of PACAP (both PACAP38 and PACAP27), when compared to any form of VIP, on cAMP production in this tissue, suggests its interaction with both PAC1 and VPAC receptors. PMID- 15800377 TI - Corticotropin-releasing factor mRNA and substance P receptor binding in the paraventricular hypothalamic nucleus, central nucleus of the amygdala, and locus coeruleus of Sprague-Dawley rats following restraint-induced stress. AB - The central mechanism of stress is poorly understood. This study was designed to examine how corticotropin-releasing factor (CRF) neurons, together with substance P (SP) receptors in the paraventricular hypothalamic nucleus (PVN), central nucleus of the amygdala (CeA), and locus coeruleus (LC), are affected by stress. Sprague-Dawley rats were restrained for 2 h. Animals were sacrificed by decapitation immediately after the 2-h restraint (the 0-h group) and 4, 24, or 48 h after restraint. Tissue sections were cut and collected on two sets of slides. Tissue sections of the first set were processed for studying CRF mRNA using 33P labeled 60-mer oligonucleotide probe. Immediately adjacent tissue sections were processed for studying SP receptor-binding capacity using 125I-SP ligand. Quantitative results showed that CRF mRNAs in the PVN were significantly up regulated at the 4- and 24-h stages, and they seemed not to be regulated by SP receptors. In addition, SP receptors in the CeA were up-regulated at the 24- and 48-h stages, whereas SP receptors were down-regulated in the LC at the same stages. In concert with the literature indicating SP antagonist's antidepressive effects, up-regulated SP receptors in the CeA might contribute to the development of stress-related depression. PMID- 15800378 TI - Altered calmodulin response to light in the suprachiasmatic nucleus of PAC1 receptor knockout mice revealed by proteomic analysis. AB - In mammals circadian rhythms are generated by a light-entrainable oscillator located in the hypothalamic suprachiasmatic nucleus (SCN). Light signals reach the SCN via a monosynaptic neuronal pathway, the retinohypothalamic tract, originating in a subset of retinal ganglion cells. The nerve terminals of these cells contain the classical neurotransmitter glutamate and the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP), and there is evidence that these two transmitters interact to mediate photoentrainment of the oscillator in the SCN. To elucidate light-provoked PACAP receptor signaling we used proteomic analysis. Wild-type mice and mice lacking the PAC1 receptor (PAC1 /-) were light stimulated at early night, and the SCN was examined for proteins that were differentially expressed using two-dimensional gel electrophoresis and identification by tandem mass spectrometry. The most striking finding, which was subsequently confirmed by Western blotting, was a significant reduction of calmodulin (CaM) in wild-type mice as compared with PAC1-/- mice. Analysis at the mRNA level by quantitative in situ hybridization histochemistry was inconclusive, indicating that a translational mechanism might be involved. The findings indicate that PAC1 receptor signaling in the SCN in response to light stimulation induces a down-regulation of CaM expression and that CaM is involved in the photic-entrainment mechanism. PMID- 15800380 TI - Endogenous neuropeptide Y prevents recurrence of experimental febrile seizures by increasing seizure threshold. AB - Febrile seizures (FSs) typically occur at the onset of fever and do not recur within the same febrile episode despite enduring or increased hyperthermia. Recurrent seizures during the same febrile episode are considered "complex," with potentially altered prognosis. A characterized immature rat model of FS was used to test the hypotheses that (1) a first FS influences the threshold temperature for subsequent ones, and (2) the underlying mechanisms involve the release and actions of the endogenous inhibitory hippocampal neuropeptide Y (NPY). Experimental FSs were induced two or three times, at 3- to 4-h intervals, and threshold temperatures measured. To determine the potential effects of seizure induced endogenous NPY on thresholds for subsequent seizures, an antagonist of the major hippocampal NPY receptor (type 2) was infused prior to induction of the second seizure. As an indicator of NPY release, NPY expression was determined 4 and 24 h later. Threshold core and brain temperatures for hyperthermic seizures were consistent with those observed during human fever. Threshold temperatures for a second and third seizure were significantly and progressively higher than those required for the first. This "protective" effect involved induction of endogenous NPY because it was abolished by the NPY antagonist. In addition, NPY mRNA expression was increased in dentate gyrus, CA3 and CA1, after an experimental FS, consistent with peptide release. Collectively these data indicate that the absence of repetitive seizures during a febrile episode involves the inhibitory actions of endogenous NPY, suggesting that the signaling cascade triggered by this peptide might provide targets for therapeutic intervention. PMID- 15800379 TI - Brain effects of chronic IBD in areas abnormal in autism and treatment by single neuropeptides secretin and oxytocin. AB - Recent research points to the connection between behavioral and gut disorders. Early adverse events are associated with inflammatory bowel disease (IBD). In animal models, maternal deprivation and social isolation predispose to gastric erosion and brain pathology. This study examined (1) brain effects of chronic gastrointestinal inflammation in a rat model of acquired IBD and (2) whether such changes are resolved by individual secretin (S) or oxytocin (OT) peptide treatment. Neurological manifestations of IBD were mapped by c-fos gene expression in male Sprague-Dawley rats (n = 10) with trinitrobenzene sulfonic acid (TNBS)-induced IBD vs controls (n = 11). IBD was characterized by moderate/severe infiltration of inflammatory cells 10 d after TNBS infusion. Age matched pairs were processed for immunocytochemical detection of Fos, expressed when neurons are stimulated. S or OT (100 mg/250 mL saline) or equivolume saline was administered iv by Alzet pump for 20 d after disease onset. Degree of resolution of colitis-induced brain activation was assessed by c-fos expression, and mean numbers of Fos-immunoreactive nuclei for each group were compared using Independent Samples T-test. Chronic IBD activated periventricular gray, hypothalamic/visceral thalamic stress axes and cortical domains, and septal/preoptic/amygdala, brain areas abnormal in autism. Single peptide treatment with S or OT did not alter the effects of inflammation on the brain. Brain areas concomitantly activated by visceral inflammation are those often abnormal in autism, suggesting that IBD could be a model for testing treatments of autism. Other single and combined peptide treatments of IBD should be tested. The clinical implications for treating autism, IBD, and concomitant sickness behaviors with peptide therapy, with or without maternal nurturing as a natural equivalent, are presented. PMID- 15800382 TI - "Local Food - Nutraceuticals": an example of a multidisciplinary research project on local knowledge. AB - Currently, dietary patterns change rapidly all over the world. Most notably, there is a fast increase in the convenience food market. Here we discuss the overall theoretical framework and strategy of an EU-funded project on local food, a common resource in many parts of the Mediterranean. Such food is often only available seasonally and is consumed either fresh (e.g. spring salads and vegetables, fruits in autumn) or in a conserved form (dried, fermented, pickled). There is an urgent need to document and analyse such local resources, which are today at the brink of disappearance. In this project, selected species were studied using a multidisciplinary approach, including strategies and methods from pharmacology, nutritional sciences and anthropology (i.e. ethnopharmacological or ethnonutritional ones). For example, all extracts were profiled using HPLC-MS, by determining their polyphenol content and using a variety of in vitro anti-oxidant assays (incl. guaiacol oxidation, xanthine oxidase inhibition, HOCl scavenging, eNOS activity). Such research also points to ways for ascertaining the intergenerational transmission of the knowledge and for sustainable development and management. Examples from field studies in southern Italy and from pharmacological studies using a variety of targets are used to illustrate the potential of such neglected resources. The wider implications of such an approach, for example, for the study of similar traditions in Central and Eastern Europe are also discussed. PMID- 15800381 TI - Microarray analysis of postictal transcriptional regulation of neuropeptides. AB - Unlike adults, kainic acid (KA)-induced status epilepticus (SE) in immature rats causes neither cell death nor recurrent spontaneous seizures. To elucidate the mechanisms of these distinct responses, transcriptional changes in neuropeptides were examined following KA-induced SE. We aimed to determine whether neuropeptides with anticonvulsant/neuroprotective properties were preferentially increased in immature rats while those with a proconvulsant/neurotoxic role were elevated to a greater extent in mature rats. We used high-density oligonucleotide gene arrays and directly compared transcriptional regulation of seven select neuropeptides at P15 and P30 over five time points. Total RNAs were isolated from hippocampi of 12 animals and pooled to hybridize to triplicate Affymetrix Genechips. Microarray results were validated by real-time quantitative RT-PCR (qRT-PCR). Independent individual RNA samples were purified for triplicate runs of qRT-PCR. Neuropeptides are significantly regulated by seizures in both immature and mature hippocampus. The magnitude of increase is significantly higher at P30 compared with that at P15, not only for neuropeptides with neurotoxic/proconvulsant properties but also for those with neuroprotective/ anticonvulsant properties. Galanin is induced at 24 h only in P30 rats. CST shows high expression in immature hippocampus and is further increased after KA-induced SE only in P15. The expression trends seen in the microarray data are confirmed by qRT-PCR for all six neuropeptides analyzed. CST might play a neuroprotective role in immature rats, and its overexpression might prevent neuronal loss after seizure in adults. Also, suppression of tachykinin and corticotropin-releasing hormone might be effective in alleviating seizure-induced neuronal damage. PMID- 15800383 TI - Flavonoids and the aging brain. AB - Like in all other organs, the functional capacity of the human brain deteriorates over time. Pathological events such as oxidative stress, due to the elevated release of free radicals and reactive oxygen or nitrogen species, the subsequently enhanced oxidative modification of lipids, protein, and nucleic acids, and the modulation of apoptotic signaling pathways contribute to loss of brain function. The identification of neuroprotective food components is one strategy to facilitate healthy brain aging. Flavonoids were shown to activate key enzymes in mitochondrial respiration and to protect neuronal cells by acting as antioxidants, thus breaking the vicious cycle of oxidative stress and tissue damage. Furthermore, recent data indicate a favorable effect of flavonoids on neuro-inflammatory events. Whereas most of these effects have been shown in vitro, limited data in vivo are available, suggesting a rather low penetration of flavonoids into the brain. Nevertheless, several reports support the concept that flavonoid intake inhibits certain biochemical processes of brain aging, and might thus prevent to some extent the decline of cognitive functions with aging as well as the development or the course of neurodegenerative diseases. However, more data are needed to assess the true impact of flavonoids on brain aging. PMID- 15800384 TI - Mediterranean food and health: building human evidence. AB - Adherence to a Mediterranean style diet affords protection from degenerative diseases such as cardiovascular disorders and cancer. Identification of the active constituents of the Mediterranean diet is crucial to the formulation of appropriate dietary guidelines. Also, research on the pharmacological properties of the "minor components" of this diet, eg vitamins and polyphenols, is very active and might lead to the formulation of functional foods and nutraceuticals. Even though in vitro data are plentiful, human studies are difficult to perform due to ethical and practical reasons. Yet, intervention trials represent the best approach to validate claims of healthful activities. This article reviews human evidence of the biological properties of olive oil and tomato constituents and illustrates a research approach by which the bioactive elements of a wild plant (Cynara cardunculus) are first studied in vitro to build biochemical evidence, then in vivo to obtain proof of their vasomodularoty activity. PMID- 15800385 TI - Nutraceuticals as anti-angiogenic agents: hopes and reality. AB - Angiogenesis, the formation of new blood vessels from preexisting vascular network is a driving force of organ development in ontogeny, is necessary for ovulation and hair growth, and is prerequisite for proper wound healing. It is also a critical mechanism of numerous diseases, the most important of which are cancer and atherosclerosis. Therefore, modulation of angiogenesis is considered as therapeutic strategies of great importance for human health. Numerous bioactive plant compounds, often referred to as nutraceuticals are recently tested for the potential clinical applications. Among the most frequently studied are resveratrol, a polyphenol present in red-wine and grape-seed, epigallocatechin-3-gallate (EGCG) from green tea and curcumin from Curcuma longa. It is also possible that components of other plants, including the constituents of local food diet may find application for modulation of angiogenesis, provided that their effectiveness will be confirmed in controlled, scientifically validated trials. PMID- 15800386 TI - Potential parapharmaceuticals in the traditional Polish diet. AB - The basic parapharmaceuticals in the Polish diet include natural anti-oxidants - bioflavonoids found in berry fruit. They were proven to have the ability to regulate genetic transcription and increase the synthesis of nitric oxide which counteracts dysfunction of the vascular endothelium. They also display anti oxidant action through the inhibiting effect on cyclooxygenase - COX-2, and increase the level of adiponectin. We have also more and more proof of the important biological role of short-chained fatty acids formed as a result of fermentation of fibre by probiotic bacteria. Through their effects on peroxisome proliferators activated receptors (PPAR), butyric and propionic acids may reduce the expression of adhesion molecules and exert anti-inflammatory action both in the gastrointestinal tract as well as systemically. PMID- 15800387 TI - Bonistein (synthetic genistein), a food component in development for a bone health nutraceutical. AB - In the discussion of the risk-benefit relation of the hormone replacement therapy (HRT) for elder women phytochemicals with estrogenic activity received a great deal of attention. Phytoestrogens are naturally occurring compounds with structural similarity to 17beta-estradiol. Especially genistein, an isoflavone most abundant in soy, possess a high and selective binding-affinity to the mammalian estrogen receptors. It has been found, that genistein exert in humans both: weak estrogenic and anti-estrogenic effects, similar to the SERMs. Consequently, it was concluded, that genistein might provide an alternative to prevent postmenopausal bone-loss and ameliorate menopausal symptoms without side effects similar to HRT. Pre-clinical experiments and results from clinical pilot studies with pure genistein confirmed its efficacy in these indications. Nevertheless, currently some open issues still exist to recommend its intake thoughtlessly. Bonistein, pure synthetic genistein developed by DSM Nutritional Products, was tested extensively in appropriate models for bone health. A battery of toxicological studies was conducted to determine safe intake levels. In the early clinical development pharmacokinetic studies were performed in healthy volunteers and in postmenopausal women. Now large-scale studies are in preparation to investigate Bonistein's efficacy in postmenopausal bone-loss and climacteric syndrome. PMID- 15800388 TI - The ethnobotanical study of local Mediterranean food plants as medicinal resources in Southern Spain. AB - We studied medicinal and food plant species, recording an extraordinary number of species and uses in Castilla-La Mancha, Murcia and Valencia in Spain. Focusing on two demographically distinct regions - Castilla-La Mancha and Lower Segura Valley. A high proportion of the flora (20 to 30 %) is known for its medicinal properties, and, interestingly, a high number of medicinal-food plants (5 to 7 %) is recorded. The concept of "Local Food" involves the whole repertory of species that characterises the local diet (incl. local cultivars and non-cultivated gathered food plants). The number of food plant species varies between 15 and 25 % of the vascular flora, and for the gathered food plants (GFP) this decreases to a 3 to 8 %. Those GFP that are also used for medicinal purposes are only 2 to 4 % of the total vascular flora. The relevant plant families are very similar in relative numbers along the different areas: Compositae, Rosaceae and Umbelliferae, followed of Boraginaceae, Liliaceae, Cruciferae, and Caryophyllaceae. Chenopodiaceae, Polygonaceae and Gramineae are less uniformly represented or in lesser numbers. The high species diversity does not imply a general dietary relevance of this particular plant family. For instance Caryophyllaceae with a relatively low number of species comprises the "collejas" (Silene vulgaris) that have shown to be the more widely consumed species. Information regarding 145 species has been recorded. Among the Gathered Food Plant Species 81 are used in medicine, in double proportion than the cultivated food plants. 61 are orally administered, in the same form as food. PMID- 15800389 TI - Antioxidant properties of Mediterranean food plant extracts: geographical differences. AB - Locally grown, wild food plants seasonally contribute a considerable portion of the daily diet in certain Mediterranean areas and it has been suggested that the beneficial effects of the Mediterranean diet on human health partly originate from the antioxidant effect of flavonoid-rich food plants. The nutrient content of most wild plants is higher than that of cultivated ones and may vary depending on the prevailing environmental conditions. Accordingly, three local Mediterranean plant foods (i.e. Cichorium intybus, Sonchus oleraceus, Papaver rhoeas) were collected in Greece (Crete), southern Italy, and southern Spain in order to assess possible differences in their in vitro antioxidant potential. The biological assays revealed diverse intra-plant specific antioxidant effects for the tested extracts ranging from no activity to almost complete protection. Furthermore, substantial differences in the polyphenol content were found for the nutritionally used part of the same plant originating from different locations. However, no clear correlations between the polyphenol content and the extracts' antioxidant activities were found. Taken together, the data suggest that certain local Mediterranean plant foods possess promising antioxidant activity and that the observed biological effects are possibly influenced by the geographically dependent environmental conditions prevailing during plant growth. PMID- 15800390 TI - Effects of plant extracts on angiogenic activities of endothelial cells and keratinocytes. AB - Numerous bioactive chemical compounds of plant origin may influence the angiogenic activity of various cell types and may thus affect the formation of blood vessels. Here we present the angiogenic effects of extracts of edible plants collected in Crete, Southern Italy and Southern Spain. Extracts have been applied to cultured human microvascular endothelial cells (HMEC-1), human umbilical vein endothelial cells (HUVEC) and human keratinocytes (HaCaT). About half out of 96 extracts exerted an inhibitory effect on HMEC-1 proliferation. Additionally, we have noted the inhibitory effects of extracts on HUVEC differentiation on a Matrigel layer. None of the extracts showed a stimulatory activity. The extract of Thymus piperella exerted moderate inhibitory effect on cobalt-chloride induced VEGF synthesis, however, CoCl(2)-induced activation of hypoxia responsive element of VEGF promoter was significantly attenuated only by extract of Origanum heracleoticum. Our study indicates that extracts of local food plants, of potential value as nutraceuticals, contain chemical compounds which may inhibit angiogenesis. Demonstration of their real influence on human health requires, however, extensive animal studies and controlled clinical investigations. PMID- 15800391 TI - Anti-inflammatory effects of extracts from some traditional Mediterranean diet plants. AB - It is believed that bioactive compounds from plant foods may have health beneficial effects and reduce the risk of chronic inflammatory diseases. In this study extracts of 121 plants typical for the traditional Mediterranean diet have been screened for their potential anti-inflammatory activities. The ability of the extracts to inhibit cytokine-stimulated, iNOS-dependent synthesis of nitric oxide in murine endothelial cells, without affecting cell viability, was the primary indicator of their anti-inflammatory properties. Based on these experiments we selected eight plant extracts for further analysis: Chrysanthemum coronarium L., Scandix pecten-veneris L., Urospermum picroides (L.) Scop. Ex F. W. Smith, Amaranthus cf. graecizans L., Onopordum macracanthum Schousboe, Eryngium campestre L., Artemisia alba Turra and Merendera pyrenaica (Pourret) Fourn. Only the effects of Onopordum macracanthum could be non-specific since the extract strongly inhibited total protein synthesis. All remaining 7 extracts decreased nitric oxide and TNFalpha synthesis in the cells of monocyte origin activated with LPS, and 4 of them significantly reduced surface expression of VCAM1 on TNFalpha-stimulated endothelial cells. All seven plant extracts decreased cytokine or LPS-stimulated iNOS mRNA levels in both cell types. Further research to identify bioactive compounds influencing intracellular signaling pathways activated by cytokines and LPS will consequently be needed in order to better understand these in vitro effects. PMID- 15800392 TI - Transcription factors as targets of the anti-inflammatory treatment. A cell culture study with extracts from some Mediterranean diet plants. AB - During the inflammatory response at least 2 transcription factors, NF-kappaB and AP-1, are involved in the altered profile of gene expression. We used human hepatoma (HepG2) and human umbilical vein endothelial cells (HUVEC) as a model system: NF-kappaB and AP-1 were activated by the proinflammatory cytokine IL-1 in the absence or presence of 21 selected plant extracts and the effect was evaluated by the electrophoretic mobility shift assay (EMSA). In both types of cells activation of NF-kappaB by IL-1 was significantly inhibited by extracts from Scandix australis and Artemisia alba, whereas extracts from Amaranthus sp., Eryngium campestre, Thymus pulegioides and Reichardia picroides elicited cell type dependent response. The IL-1-induced AP-1 activation was diminished by extracts from Scandix australis, Amaranthus sp. and Artemisia alba more potently in HUVEC, while extracts from Urospermum picroides and Scandix pecten-veneris in HepG2 cells. Inhibitory activities of plant extracts towards cytokine activated NF-kappaB and AP-1 depend to some extent on the order of addition of IL-1 and plant extract to the cell culture, but the mechanism of action of extract components is not clear: although plant polyphenols may participate they are unlikely to be the only mediators, and MAP kinases were found generally not involved in down-regulation of transcription factors activity by plant extracts. PMID- 15800393 TI - Dietary intake and biochemical risk factors for cardiovascular disease in two rural regions of Crete. AB - The aim of the present study was to identify any possible protective effects of wild greens on certain biochemical cardiovascular disease (CVD) risk factors in elderly women leaving in rural Crete. For the needs of the study a region with high consumption of wild greens (Avdou) and a region with low consumption (Anogia) were identified and a representative population from each area was recruited (n= 37 and n=35 respectively). Serum lipids and fibrinogen levels, total antioxidant capacity (TAC), soluble intercellular adhesion molucule-1 (sVCAM-1), soluble vascular adhesion molucule-1 (s-ICAM-1) and haematological factors were measured in both regions during winter and summer time when wild green plants consumption is high and low, respectively. Regarding classic lipid risk factors for arteriosclerosis no significant different between the two regions were detected. TAC was found higher in Avdou compared to Anogia during winter screening, but null in both regions during summer. Fibrinogen was found higher in Avdou compared to Anogia while no differences between the two regions were detected for sVCAM-1 and s-ICAM-1 in both screenings. The significantly higher TAC and fibrinogen values detected in Avdou could be attributed to the higher wild green consumption in that region since recent data indicated that their content in vitamin C and E as well as flavonoids is particularly high. PMID- 15800394 TI - The protective ability of the Mediterranean plant extracts against the oxidative DNA damage. The role of the radical oxygen species and the polyphenol content. AB - The polyphenol plant extracts content seems to be responsible for the scavenging activity of the reactive oxygen species (ROS), resulting in protection against DNA damage induced by the oxidative stress. This assumption was verified analyzing the effect of six Mediterranean plant extracts (Crepis vesicaria L, Origanum heracleoticum, Scandix australis L, Amaranthus sp., Scolymus hispanicus L, Thymus piperella L) on the oxidative DNA damage induced in lymphocytes by H(2)O(2) in relation to the polyphenolic content and the lymphocyte scavenging ability of ROS. The comet assay was used to evaluate oxidative DNA damage and the polyphenol content was analyzed by the Folin-Ciocalteu method. The fluorescence resulting from oxidation of ROS-sensitive dye, dihydrorofluorescein (DHF), was utilized as indicator of the ROS level. Pretreatment with all plant extracts produced the dose-dependent increase in the DNA protection up to the 0.2 microg/ml polyphenol content and the decrease above that dose. Only the Thymus piperella, similarly to quercetin, showed a strong positive correlation between the DNA protection and the polyphenol content, but negative correlation with ROS formation. In conclusion, the DNA protective ability of plant extracts seems to be related to the low polyphenol concentration and only to certain extent depends on the polyphenol ROS scavenging activity. PMID- 15800395 TI - The estrogenic status and the COMT genotype of female blood donors influence the protective ability of the Mediterranean plant extracts against the hydrogen peroxide-induced DNA damage in lymphocytes. AB - The extent of oxidative DNA damage in lymphocytes can be used as a biomarker of the level of oxidative stress in the body. The comet assay has been widely used to measure such damage. The aim of our study was to evaluate: i) the extent of the oxidative DNA damage in lymphocytes isolated from blood of female donors taken in early and late follicular phases [low (LE) and high (HE) concentration of 17beta-estradiol, respectively], ii) the susceptibility of these lymphocytes to hydrogen peroxide exposure, and iii) the protective ability of five plant extracts against the hydrogen peroxide-induced DNA damage. The effect of the catechol-O-methytransferase genotype (wild COMT H/H and mutated homozygote COMT L/L) of female donors was also analyzed. The amount of endogenous DNA damage was higher in HE lymphocytes as compared with LE ones, independently of the genotype. When lymphocytes were stratified by COMT genotype, the level of DNA damage was higher in L/L donors. The protective effect of pretreatment with plant extracts (1 and 10 micro g/ml for 1 h) against the H(2)O(2) (25 microM, 5 min. at 4 degrees C)-induced oxidative DNA damage was observed only in H/H HE lymphocytes. In contrary, the plant extract pre-incubation enhanced the DNA damage in L/L HE lymphocytes. The plant extracts alone did not induce the DNA damage. The results showed that concentration of the circulating 17beta-estradiol influenced the extent of endogenous oxidative DNA damage while the beneficial or hazardous effects of the plant extracts might depend on the COMT genotype and the estrogen level. PMID- 15800396 TI - Gastroprotective effects of flavonoids in plant extracts. AB - The purpose of this paper is to overview the relations between plant-originated substances and their bioactivity measured in terms of antioxidant, cytoprotective and antiulcer activities. In addition, we assessed whether these compounds are capable of affecting the gastric mucosal lesions induced by absolute ethanol applied intragastrically (i.g.). The following plant-originated flavonoid substances were considered; Solon (Sophoradin extract), Amaranth seed extract, grapefruit-seed extract (GSE) and capsaicin (extract of chilly pepper). The area of gastric mucosa lesions and gastric blood flow were measured in rats with ethanol-induced lesions without (control) and with one of the tested substances without and with capsaicin denervation of afferent nerves or administration of L nitro-arginine (L-NNA), an inhibitor of nitric oxide synthase (NOS). Male Wistar rats, weighing 180-220 g fasted for 24 h before the study where used 100% ethanol was applied i.g. to induce gastric lesions, whose area was determined by planimetry. Gastric blood flow was assessed using electrolytic regional blood flowmeter. All tested plant-originated substances afforded gastroprotection against ethanol-induced damage and this was accompanied by increase in gastric microcirculation, both changes being reversed by pretreatment with neurotoxic dose of capsaicin or by pretreatment with L-NNA. We conclude that plant originated flavonoid substances are highly gastroprotective probably due to enhancement of the expression of constitutive NOS and release of NO and neuropeptides such as calcitonin gene related peptide (CGRP) released from sensory afferent nerves increasing gastric microcirculation. PMID- 15800397 TI - Lack of combination hepatocarcinogenicity of harman, norharman and amitrole when given with NaNO2 in the rat. AB - N-nitrosocompounds, which induce cancers in various organs, may be formed endogenously with intake of amino compounds such as secondary amines and sodium nitrite (NaNO(2)) in combination. The present study was performed to investigate whether three amino compounds, 1-methyl-9H-pyrido[3,4-b]indole (harman), 9H pyrido[3,4-b]indole (norharman) and 2-amino-1,3,4-triazole (amitrole), might be converted in vivo to compounds capable of promoting hepatocarcinogenesis when given with NaNO(2). However, in an 8-week model, no modifying potential was evident in terms of numbers and areas of putative preneoplastic glutathione S transferase placental form (GST-P)-positive foci in any of the groups receiving paired treatments. These results demonstrate that combinations of harman, norharman and amitrole with NaNO(2) lack promoting effects for liver carcinogenesis in our medium-term bioassay system. PMID- 15800398 TI - Effects of maternal exposure to a low dose of diethylstilbestrol on sexual dimorphic nucleus volume and male reproductive system in rat offspring. AB - Diethylstilbestrol (DES) was administered subcutaneously at 0.5, 1.5 or 4.5 microg/kg/day (DES 0.5, 1.5 and 4.5 groups, respectively) to pregnant SD rats daily on days 7-21 of gestation, to investigate its effects on the development and functions of the reproductive system in their male offspring. Of the 10 pregnant rats in the DES 4.5 group, only 1 delivered, and this rat could not suckle the pups. Rat pups in the DES 0.5 and 1.5 groups were autopsied at 1, 3, 6 and 15 weeks after birth. The testosterone concentrations in the DES 1.5 and 0.5 groups at 6 weeks were significantly decreased and the plasma LH concentrations were not altered. In the DES 1.5 group, DES treatment did not change the volume of the sexually dimorphic nucleus in the preoptic area (SDN-POA) in the male offspring, although this dose of DES increased the volume of SDN-POA in female offspring. The DES treatment altered frequencies in the cycles of the seminiferous tubules, and suppressed histological maturation in the epididymis and the prostate weight. These observations indicate that prenatally administered DES impairs testicular endocrine function continuously as well as pituitary function, but the induced low level of testosterone disrupts spermatogenesis and permanently inhibits the morphological development of epididymis and prostate. PMID- 15800399 TI - Phototoxicity study of a ketoprofen poultice in guinea pigs. AB - Ketoprofen has been reported to have such side effects as photosensitive dermatitis in humans (The Ministry of Health, Labour and Welfare, 2001). In the present study, as part of a safety evaluation of Miltax, an application drug containing ketoprofen, phototoxicity of Miltax was examined in guinea pigs. In the present skin phototoxicity study, Miltax was applied for 12 hr. Ultraviolet (UV) rays were irradiated to examine whether or not Miltax elicited skin reaction. Two kinds of UV-A plus UV-B dual irradiation and UV-A single irradiation were used for the elicitation. With UV-A plus UV-B dual irradiation on the Miltax application site, no skin reaction was observed at UV irradiation side in any animals, in contrast to the case of the positive control article, 8 methoxypsoralen (8-MOP). Similar results were obtained with UV-A single irradiation. From these results, Miltax that contained ketoprofen did not show any skin phototoxicity in the guinea pig. PMID- 15800400 TI - Susceptibility of newborn rats to hepatotoxicity of 1,3-dibromopropane and 1,1,2,2-tetrabromoethane, compared with young rats. AB - Newborn rat studies were conducted with oral administration of 1,3-dibromopropane (DBP) and 1,1,2,2-tetrabromoethane (TBE) from postnatal Days 4 to 21 to allow comparison of NOAELs and unequivocally toxic levels with those from 28-day young rat studies starting at 5-6 weeks of age. The unequivocally toxic level was estimated by our specified criteria, requiring simultaneous change of organ weights, histopathology, some biochemical parameters and body weights, because in this study only hypertrophy of hepatocytes was observed as a major histopathological change. DBP caused centrilobular hypertrophy of hepatocytes with alteration in biochemical parameters, as well as lowering of body weights, regardless of sex, in both newborn and young rats. NOAELs and unequivocally toxic levels were considered to be 50 and 150 mg/kg/day in newborn rats and 10 and 250 mg/kg/day in young rats, respectively. In the newborn rat study of TBE, some hepatic effects observed at the top dose of 50 mg/kg were not considered adverse because of the lack of histopathological changes. Significant lowering of body weight was noted at 200 mg/kg in the dose-finding study but histopathological data were not available. In the young rat study, there was no definite toxicity at 6 mg/kg and hypertrophic changes in liver and thyroids without body weight change occurred at 200 mg/kg. There were no clear sex differences in both the newborn and young rat studies. NOAELs were considered to be 50 and 6 mg/kg/day in newborn and young rats, respectively, but unequivocally toxic levels for both rats could not be estimated. Abnormalities of external and sexual development and reflex ontogeny in the newborn were not observed with either chemical. Based on these results, it can be concluded that the target organ of DBP and TBE is the liver in both newborn and young rats, and that while the doses at which toxic signs began to appear are higher in newborn rats, those causing clear toxicity may be paradoxically lower in the newborn case. PMID- 15800401 TI - Roles of mu-opioid receptors in development of tolerance to diisopropylfluorophosphate (DFP). AB - Anatomical evidence indicates that cholinergic and opioidergic systems are co localized and acting on the same neuron. However, the regulatory mechanisms between cholinergic and opioidergic system have not been well characterized. In the present study, the potential involvement of mu-opioid receptors in mediating the changes of toxic signs and muscarinic receptor binding after administration of irreversible anti-acetylcholinesterase diisopropylfluorophosphate (DFP) was investigated. DFP (1 mg/kg/day, subcutaneous injection, s.c.)-induced tremors and chewing movements were monitored during the 28-day treatment period in mu-opioid receptor knockout and wild type mice. Autoradiographic studies of total, M1, and M2 muscarinic receptors were conducted using [(3)H]-quinuclidinyl benzilate, [(3)H]-pirenzepine, and [(3)H]-AF-DX384 as ligands, respectively. DFP-induced tremors in both mu-opioid receptor knockout and wild type mice showed tolerance development. However, DFP-induced tremors in mu-opioid receptor knockout mice showed delayed tolerance development than that of DFP-treated wild type controls. DFP-induced chewing movements in both mu-opioid receptor knockout and wild type mice failed to show development of tolerance after four weeks of treatment. M2 muscarinic receptor binding of DFP-treated mu-opioid receptor knockout mice was significantly decreased than that of the DFP-treated wild type controls in the striatum, but not in the cortex and hippocampus. However, there were no significant differences in total and M1 muscarinic receptor binding between DFP treated mu-opioid receptor knockout and wild type mice in the cortex, striatum and hippocampus. These studies indicate that mu-opioid receptors play an important role through the striatal M2 muscarinic receptors to regulate the development of tolerance to DFP-induced tremors. PMID- 15800402 TI - Investigation of a hepatotoxicity screening system in primary cell cultures - "what biomarkers would need to be addressed to estimate toxicity in conventional and new approaches?". AB - High throughput toxicological estimation is required for safety evaluation in the early stage of drug discovery. In this context, establishment of an in vitro screening system reflecting in vivo toxicity is demanded for earlier safety assessment. We investigated LDH release and mitochondrial respiration (WST-1 reduction assay; WST-1) to detect cytotoxicity, morphological evaluation, and proteomics for estimating the reliable and sensitive biomarkers by using rat primary hepatocytes exposed to the compounds (acetaminophen, amiodarone, tetracycline and carbon tetrachloride) that are known to induce hepatotoxicity. In LDH release, no significant difference was detected between the control and compound exposed cells after exposure for 3 or 6 hr, but a dose-dependent increase was observed after exposure for 24 hr. Regarding the WST-1 assay, a dose dependent reduction was detected after exposure for 6 and 24 hr to all of the compounds evaluated. In the proteomics analysis, 31 candidate proteins were identified from among the 103 demonstrating altered expression spots after exposure to acetaminophen. It was concluded that the cytotoxicity was detected earlier by measuring WST-1 than by measuring LDH release because the reduction of mitochondrial respiration is an expressions of earlier toxicity for cellular function, while the measured increase in the LDH release occurs after the failure of the cell membrane. Mitochondrial respiration ability was a useful parameter for cytotoxicity in in vitro hepato-toxicity screening, as cytotoxicity can be detected during the early stage of exposure. In addition to the conventional biomarkers, several protein biomarkers which relate to oxidative stress and metabolism-regulation were detected. Further comprehensive analysis of defined proteins would be necessary to estimate the more sensitive toxicology biomarker. PMID- 15800405 TI - Metalloporphyrins for the treatment of neonatal jaundice. AB - PURPOSE OF REVIEW: To evaluate the safety and efficacy of metalloporphyrins for the treatment of neonatal hyperbilirubinemia. RECENT FINDING: Since the 1980s and 1990s, there have been no publications on trials to determine the efficacy of metalloporphyrins in the treatment of neonatal jaundice. In the past year, a single case report was presented on the compassionate use of tin mesoporphyrin in a very low birth weight infant with intrauterine growth retardation who did not respond to phototherapy. Subcutaneous administration of a single dose of tin mesoporphyrin at 46 hours of life was associated with a greater than 25% reduction in serum bilirubin. This further supports existing evidence that tin mesoporphyrin is efficacious in lowering bilirubin production. In the laboratory, most metalloporphyrins were shown to induce heme oxygenase, and in addition, metalloporphyrins modulate cardiac cell function in vitro. These observations suggest that the therapeutic benefits may be obviated if such considerations hold true in humans. SUMMARY: Recent case reports and previous evidence from larger clinical trials conducted in Greece and Argentina in the 1980s and 1990s demonstrate that tin mesoporphyrin is useful in the treatment of neonatal jaundice. Its long-term safety is not well understood but will be important to determine, before its widespread or prophylactic use in neonates with hyperbilirubinemia can be recommended. PMID- 15800406 TI - Feeding strategies and necrotizing enterocolitis. AB - PURPOSE OF REVIEW: To update clinical trials concerning feeding strategies or feeding interventions to prevent necrotizing enterocolitis (NEC). RECENT FINDINGS: The overall incidence of NEC remains unchanged. Several studies have shown that it is safe to use small feeding volumes early in life even in selected circumstances, such as feeding during the use of indomethacin to treat symptomatic patent ductus arteriosus. Although descriptive case reports have linked some feeding interventions such as thickened feedings to NEC, there is no evidence to establish a causal relation. Interestingly, one report showed that neither the presence of-nor characteristics of-gastric residuals was helpful to identify those infants who subsequently develop NEC. Numerous studies in animals show that vascular and host defense responses are not adequate in the immature gut. A few small clinical trials showed that various feeding strategies may alter these responses in preterm infants. However, larger clinical trials around these issues await further characterization of the mechanisms that regulate these two aspects of gastrointestinal function with respect to feeding. SUMMARY: Studies continue to demonstrate how fragile the immature gut is, but further work directed to understanding the mechanisms that regulate gastrointestinal responses to feeding is needed to design meaningful interventions for large prospective trials. PMID- 15800407 TI - Strategies for preventing bronchopulmonary dysplasia. AB - PURPOSE OF REVIEW: Neonatologists and pulmonary biologists have long sought preventive treatments for bronchopulmonary dysplasia (BPD). The purpose of this review is to highlight recent reports of a number of potential treatments intended to prevent BPD and to discuss the controversies surrounding preventive strategies. RECENT FINDINGS: The evolution of BPD from a disorder of pulmonary injury affecting moderately preterm infants, to one characterized by a developmental pulmonary arrest among survivors of extreme prematurity has important implications for BPD prevention. Recent recognition that the pathogenesis of BPD might have prenatal origins raises new challenges and opportunities for studies of BPD prevention; however, most current preventive strategies for BPD focus on respiratory management. Neither past nor current clinical trials have shown a conclusive benefit of a single preventive treatment strategy. Promising but still largely unproven preventive respiratory treatments include: high frequency oscillatory ventilation, permissive hypercapnea, and inhaled nitric oxide. Observational and recent laboratory data support the need for randomized clinical trials of continuous positive airway pressure versus mechanical ventilation. Additionally, clinical trials are needed to address the deficit in our knowledge of the potential benefits and risks of postnatal low dose corticosteroid treatment. Further study of superoxide dismutase, inositol, and alpha-1 proteinase inhibitor also are warranted on the basis of recent clinical trials or meta-analyses. SUMMARY: Only Vitamin A has proven a safe and effective preventive treatment for BPD. Additional studies of respiratory technologies, management strategies, and protective molecules are needed. Directed cytokine and genetic therapies are on the horizon. PMID- 15800408 TI - Current controversies in the pathophysiology and prevention of sudden infant death syndrome. AB - PURPOSE OF REVIEW: To examine recent research relevant to sudden infant death syndrome (SIDS) to determine whether there is a place for home monitoring in the care of children believed to be at risk. RECENT FINDINGS: Current SIDS research has focused on the genetics of SIDS, brainstem abnormalities and arousal failures, the effects of tobacco smoke and other environmental agents, the role of infectious diseases, and prenatal factors that may contribute to SIDS. Investigations have suggested that there are infants who appear to respond less effectively when challenged by certain environmental or infectious agents. These infants have blunted responses to stress and diminished arousal to hypoxemia, in part because of failures in genetically determined brainstem function. It is unclear at this time whether home monitoring would offer protection in all circumstances, but it may be helpful in certain patients. SUMMARY: There appears to be progress in understanding the causes of SIDS. As additional studies emerge, the optimal approaches to care will become more apparent, with home monitoring one of the possible interventions. PMID- 15800409 TI - Ethical dilemmas in the care of the most premature infants: the waters are murkier than ever. AB - PURPOSE OF REVIEW: Summarize the literature relevant to ethical issues surrounding decisions to provide intensive care to extremely premature newborns. RECENT FINDINGS: A Texas Supreme Court decision and a position paper are noteworthy for health professionals participating in management decisions with families at risk for extremely preterm delivery. SUMMARY: In Miller v HCA, the Millers sued the Hospital Corporation of America for resuscitating their approximately 23-week gestation daughter against their wishes. The baby survived with severe neurodevelopmental disabilities. They were awarded $59.9 million in a jury trial. However, the judgment was reversed by the court of appeals, which ruled that parents have no right to withhold urgently needed life-sustaining medical treatment from children with non-terminal impairments, deformities, or disabilities, regardless of their severity. The Supreme Court of Texas upheld that ruling, but reasoned that parents have no right to refuse resuscitation of extremely premature infants prior to birth because they cannot be fully evaluated until birth; therefore, decisions before birth could not be fully informed. Robertson (Hasting Center Report 2004) supports precluding parental refusal of resuscitation before birth. He argues that parents have a right to withhold or withdraw medical treatment from a non-terminally ill child, but only if the child will lack capacity for symbolic interaction. Such severe limitation of quality of life concerns in decision making for extremely premature newborns is inconsistent with current published guidelines, the positions of noted bioethicists, and the practice of many neonatologists. Further, the additional information attained by initiating intensive care in the most premature infants does not justify doing so without parental consent. PMID- 15800411 TI - Cellular and molecular derangements in acute tubular necrosis. AB - PURPOSE OF REVIEW: Acute tubular necrosis secondary to ischemic acute renal failure remains a common clinical problem with serious consequences and unsatisfactory therapeutic options. The purpose of this review is to summarize recent advances that have provided an improved understanding of the underlying cellular and molecular derangements, and have resulted in the design of novel therapeutic approaches. RECENT FINDINGS: Sophisticated morphologic studies have identified apoptosis and vascular changes as significant novel findings in human acute tubular necrosis. Promising roles for inhibitors of apoptosis have been proposed. Activation of tubuloglomerular feedback, previously thought to contribute to acute tubular necrosis, has now emerged as a potentially beneficial phenomenon. The role of reactive oxygen molecules has been further elucidated, and novel antioxidants and iron chelators have been identified. Genome-wide screening techniques have identified the molecular mechanisms underlying the regeneration and repair processes, and have provided clues towards accelerating recovery from acute renal failure. An improved understanding of the role of inflammation has suggested strategies to target this previously underappreciated aspect of acute tubular necrosis. SUMMARY: The cellular and molecular tools of modern science have provided critical new insights into the roles of apoptosis, oxidant and iron-mediated injury, endothelial changes, regeneration, and the inflammatory response in the pathogenesis of acute tubular necrosis. Novel strategies that modulate these pathways hold tremendous promise for the proactive treatment of human acute renal failure. PMID- 15800412 TI - Hemolytic uremic syndrome; pathogenesis, treatment, and outcome. AB - PURPOSE OF REVIEW: The hemolytic uremic syndrome (HUS) is the most common cause of acute renal failure in infants and young children, and is a substantial cause of acute mortality and chronic morbidity. It is therefore relevant and appropriate that pediatricians remain familiar with the various subsets of the disease including its classification, management, and outcome. RECENT FINDINGS: This review will focus on recent information relative to epidemiology, pathogenesis, treatment, and outcome. It will include some of the newer associations between HUS and a variety of infections, including, but not limited to E. coli 0157:H7 (Shiga toxin-mediated) HUS, as well as the ever-increasing number of associations between HUS and a variety of drugs. It will review some of the newer therapies for the more common subsets, but will acknowledge that choosing evidence-based therapies is often limited by our incomplete understanding of the various pathogenic cascades, and that with the possible exception of Shiga toxin-mediated HUS(D+HUS), long-term outcome information is often limited by small numbers and limited follow-up. SUMMARY: This review should provide a framework for making the proper diagnosis, implementing appropriate treatment, and advising the family about anticipated outcome. PMID- 15800413 TI - Renal replacement therapy and acute renal failure. AB - PURPOSE OF REVIEW: Acute renal failure (ARF) is a syndrome that occurs when there is a sudden decline in the glomerular filtration rate. The purpose of this review is to examine new developments and clinical applications of renal replacement therapies including hemodialysis, continuous renal replacement therapy, the bioartificial kidney, and peritoneal dialysis in the management of this complicated syndrome. RECENT FINDINGS: New developments in hemodialysis include in-line hematocrit monitoring and improved biocompatible dialyzer membranes. While recent studies indicate that increased delivery of dialysis improves the outcome of patients with ARF, the optimal regimen of intermittent dialysis or continuous renal replacement therapy remains to be determined. The bioartificial kidney, combining hemofiltration with a device containing human tubular cells, is currently in clinical trials and represents another alternative in the management of ARF. In peritoneal dialysis, new solutions using icodextrin may improve fluid removal and blood pressure. SUMMARY: The optimal choice of renal replacement therapy depends on many factors. Use of new options in renal replacement therapy and early initiation of dialysis may help to improve survival and outcome of patients with ARF. PMID- 15800414 TI - Advances in the pathogenesis and management of hypertensive crisis. AB - PURPOSE OF REVIEW: This review summarizes recent insights into to the pathophysiology of hypertensive crisis with the emphasis on newly discovered molecular mechanisms underlying hypertension and also updates current therapeutic options for treating hypertensive crisis in children. RECENT FINDINGS: There is growing evidence that the renin-angiotensin system plays a key role in the pathogenesis of hypertensive crises. Recent studies have shown that oxidative stress and factors affecting endothelial function are also important. Treatment of hypertensive crisis still focuses on lowering of blood pressure in an expeditious but safe manner. There is growing experience with IV nicardipine, which is becoming a viable alternative to sodium nitroprusside in children. SUMMARY: Current knowledge of hypertensive crisis emphasizes the need for additional animal and translational studies with the goal of identifying the underlying molecular pathogenesis and developing new therapies to optimize future treatment of hypertensive emergencies. PMID- 15800416 TI - New drug therapies for diseases in children: from atypical antipsychotics to antivenom for snakebites. PMID- 15800415 TI - Future strategies in the treatment of acute renal failure: growth factors, stem cells, and other novel therapies. AB - PURPOSE OF REVIEW: Acute renal failure remains a significant cause of morbidity and mortality in adults and children. Despite advances in understanding the pathophysiology of acute renal failure, little progress has been made in its treatment. This review assesses the recent data on current and promising new therapies for acute renal failure. RECENT FINDINGS: The first section of the review describes the recent therapies that have been used in humans, all of whom have been adults. The second section evaluates the use of agents given in experimental animal models during or after the onset of acute renal failure. The third section describes the many animal studies using therapies before the onset of experimental renal failure. The final section discusses how the emerging field of stem cell research might be used to treat acute renal failure. SUMMARY: Among the recent studies in humans, the most intriguing have been the use of atrial natriuretic peptide in patients with nonoliguric renal failure and isotonic sodium bicarbonate infusions to prevent radiocontrast medium-induced renal failure. Among the agents used in animal studies, those with the greatest potential were hepatocyte growth factor and ethyl pyruvate, because they seem to protect against or accelerate recovery from acute renal failure after the renal insult. Finally, stem cell therapy may someday offer the best option for recovery from acute renal failure. PMID- 15800417 TI - Update on pediatric resuscitation drugs: high dose, low dose, or no dose at all. AB - PURPOSE OF REVIEW: Pediatric resuscitation has been a topic of discussion for years. It is difficult to keep abreast of changing recommendations, especially for busy pediatricians who do not regularly use these skills. This review will focus on the most recent guidelines for resuscitation drugs. RECENT FINDINGS: Three specific questions will be discussed: standard dose versus high-dose epinephrine, amiodarone use, and the future of vasopressin in pediatric resuscitation. The issue of using high-dose epinephrine for cardiopulmonary resuscitation refractory to standard dose epinephrine has been a topic of debate for many years. Recently, a prospective, double-blinded study was performed to help settle the debate. These results will be reviewed and compared with previous studies. Amiodarone is a medication that was added to the pediatric resuscitation algorithms with the most recent recommendations from the American Heart Association in 2000. Its use and safety will also be discussed. Another topic that is resurfacing in resuscitation is the use of vasopressin. Its mechanism and comparisons to other agents will be highlighted, although its use in the pediatric patient has not been thoroughly studied. SUMMARY: Pediatric resuscitation is a constantly evolving subject that is on the mind of anyone taking care of sick children. Clinicians are continually searching for the most effective methods to resuscitate children in terms of short- and long-term outcomes. It is important to be familiar with not only the agents being used but also the optimal way to use them. PMID- 15800418 TI - Toxicology and overdose of atypical antipsychotic medications in children: does newer necessarily mean safer? AB - PURPOSE OF REVIEW: Atypical antipsychotic medications (second-generation antipsychotics) have been increasingly used in the treatment of a number of psychotic disorders since their introduction in 1988, with the newest medication introduced in 2002. Justification for their use includes claims of equal or improved antipsychotic activity over first-generation antipsychotics, increased tolerability, and decreased side effects. However, there are still significant adverse effects and toxicities with this class of medications. Toxicologic exposures and fatalities associated with atypical antipsychotics continue to increase in the United States, with 32,422 exposures and 72 deaths in 2003. There have also been Food and Drug Administration warnings in the past year about how some atypical antipsychotics have been marketed to minimize the potentially fatal risks and claiming superior safety to other atypical antipsychotics without adequate substantiation, indicating the toxicologic potential of these agents may be underestimated. RECENT FINDINGS: Continued research to evaluate adverse effects and tolerability of atypical antipsychotics compared with first generation antipsychotics and each other is reviewed. This article also reviews the pharmacodynamics, pharmacokinetics, and drug interactions with these medications. New therapeutic monitoring recommendations for this class of medications have also been proposed. Finally, clinical toxicity in overdose and management are reviewed. SUMMARY: While new atypical antipsychotic medications may have a safer therapeutic and overdose profile than first-generation antipsychotic medications, many adverse and toxic effects still need to be considered in therapeutic monitoring and overdose management. PMID- 15800419 TI - Antivenom therapy for snakebites in children: is there evidence? AB - PURPOSE OF REVIEW: A new Fab fragment antivenom (CroFab) for the treatment of crotaline envenomation, the predominant venomous snakebite in the United States, has drastically changed snakebite management since its release in December 2000. This review examines the evidence supporting the use of CroFab, with particular attention on the pediatric population. RECENT FINDINGS: The published experience with CroFab in humans consists of six studies and some case reports. These publications demonstrate that CroFab is highly efficacious in treating both the local and systemic toxic effects of crotaline envenomation. They identify an important phenomenon of recurrent or delayed toxicity in some patients. The studies report a very low incidence of acute or delayed hypersensitivity reactions to the antivenom. They suggest comparable efficacy and safety of CroFab in the pediatric population as in adults. SUMMARY: Based on limited data, CroFab has been shown to be a safe and efficacious antivenom for use in children as well as adults. Further studies are needed to refine our understanding of its efficacy, safety, indications, and dosing. PMID- 15800420 TI - An update of N-acetylcysteine treatment for acute acetaminophen toxicity in children. AB - PURPOSE OF REVIEW: Acetaminophen poisoning accounts for a disproportionate percentage of all toxic ingestions, and can be life-threatening. This article reviews the mechanism and presentation of acetaminophen toxicity, as well as its treatment, including current thinking and treatment recommendations. RECENT FINDINGS: N-acetylcysteine acts to detoxify acetaminophen in several ways, but primarily by increasing the synthesis and availability of glutathione, which binds and inactivates the highly reactive and hepatotoxic acetaminophen metabolite N-acetyl-p-benzoquinoneimine. The US Food and Drug Administration has approved an intravenous formulation of N-acetylcysteine, thus allowing the treatment time to be decreased from the 72 hr most commonly used for the oral regimen, to only 20 hr. This comes after many years of accepted intravenous N acetylcysteine use in Europe and Canada, and much controversy as to the superiority of both treatments. This review summarizes this controversy, and offers a framework to develop a safe treatment plan that has the optimal outcome for the patient, as well as reflecting knowledge of the potential caveats at work. It describes side effects of N-acetylcysteine treatment, as well as relative indications to choose one route of treatment over the other. SUMMARY: Acetaminophen can lead to irreversible liver damage and even death in acute overdose. Outcome is related to the swiftness in which the antidote (N acetylcysteine) is provided. In the United States, there are now available both the oral and intravenous forms of N-acetylcysteine, and pros and cons exist for each. With brisk and adequate treatment using either route, recovery can be complete, and liver function can be restored. PMID- 15800421 TI - Update on child maltreatment with a special focus on shaken baby syndrome. AB - PURPOSE OF REVIEW: The many forms of child maltreatment exact an unacceptably heavy toll on our children. Recent work exploring the risk factors for child abuse and neglect, long-term consequences of child maltreatment, and potential for primary prevention will be reviewed. In the past year there have been many notable contributions in the field of shaken baby syndrome that have deepened our understanding of this devastating form of child abuse. RECENT FINDINGS: Evidence of the long-term sequelae of child abuse and neglect continues to accumulate, but with increased understanding of the clinical manifestations of child maltreatment has come greater ability to intervene early and perhaps improve long-term outcomes. Although the effectiveness of some home visiting programs to prevent child maltreatment is coming into question, rigorous review of program components is helping to improve quality. There remain controversies in the diagnosis of shaken baby syndrome but several carefully done case-series are clarifying our knowledge of the myriad presentations of this form of non-accidental head trauma. SUMMARY: Pediatricians have an important role to play in the prevention and recognition of child maltreatment. PMID- 15800422 TI - Update on Munchausen syndrome by proxy. AB - PURPOSE OF REVIEW: Munchausen syndrome by proxy (MBP) is a complicated form of child maltreatment. Difficulties remain in properly defining the condition, as well as in detection and differentiation from organic illness. This review will discuss the epidemiology and diagnosis of MBP, as well as the role of the physician in sorting out these cases. RECENT FINDINGS: Several recent case studies, including two in which children were diagnosed with celiac disease, add to our knowledge of the protean manifestations of MBP. There is growth in our understanding of how sudden infant death syndrome (SIDS) and the symptom complex seen in acute life-threatening events (ALTEs) may in fact represent manifestations of MBP. Recent legal issues in the United Kingdom pose concern for all physicians engaged in child protection work. SUMMARY: In spite of these challenges, the high mortality and recidivism rates associated with MBP make it imperative that pediatricians be familiar with the condition, the subtle signs and symptoms with which it may present, and methods to best protect the children in their care. PMID- 15800423 TI - Update on childhood sexual abuse. AB - PURPOSE OF REVIEW: Childhood sexual abuse (CSA) is a problem of epidemic proportion affecting children and communities worldwide. This review will examine the most recent published CSA literature emphasizing not only new findings in diagnosis, management, and prevention, but also practice implications for the medical and community-based provider. Sexual assault and rape, though forms of sexual abuse, are broader than the scope of this article and not discussed in detail at this time. RECENT FINDINGS: Prevalence rates for CSA are difficult to obtain. To identify populations at greatest risk and tailor treatment and prevention initiatives, consistent methods for defining and reporting cases of CSA are needed. Appreciating the quality and quantity of adverse health outcomes associated with CSA has helped clinicians and community providers expand and develop treatment programs. The physical and psychological late effects of abuse can be permanent and life threatening and the primary care provider should assess for mental health problems as well as physical maladies in all routine examinations. Children do not often disclose experiences of abuse for months to years and caregivers must be cognizant of the medical and behavioral indicators that signal early abuse. Child advocacy centers are community-based programs that incorporate medical, psychological, and legal support services for children and their families. This therapeutic model is based on a multidisciplinary team assessment aimed at decreasing stress, providing adequate protection and services for the child, and optimizing chances for a successful legal outcome. SUMMARY: While prevention remains the ultimate goal, improved awareness and education for communities and professionals alike is required to ensure appropriate and quality care for all children who are sexually abused. PMID- 15800424 TI - Update: attention deficit/hyperactivity disorder in the primary care office. AB - PURPOSE OF REVIEW: Attention-deficit/hyperactivity disorder (AD/HD) affects 7.5% of children, making it among the more common behavioral disorders of childhood. Pediatricians increasingly are expected to recognize AD/HD, as well as diagnose and manage it in the primary care setting. This article reviews recent developments in the care of the pediatric AD/HD patient, with emphasis on information enhancing primary care management. RECENT FINDINGS: Studies published in 2004 provide evidence to guide the treatment of AD/HD. The AD/HD literature continues to support the important role of genetics in its etiology. The absence of universal genetic or neuroimaging findings indicates that history from multiple sources and physical exam remain the standard diagnostic method. Comorbid medical problems, such as sleep disruption and growth suppression, continue to be better understood in the setting of AD/HD, as do the substantial impacts of comorbid learning and psychiatric disorders. Despite great interest in alternative, nonstimulant and behavioral treatments, methylphenidate and amphetamine-based medications remain the mainstay of AD/HD intervention. SUMMARY: AD/HD is a common medical condition with implications for long-term safety and life function, such as academic success, accident occurrence, and drug use. Identification and treatment is increasingly based in the primary care office, where children must be monitored for co-occurring disorders and referred for additional supports when necessary. Tools and guidelines provided by the American Academy of Pediatrics (AAP) provide a framework for consistent and competent AD/HD care supported by current evidence. PMID- 15800425 TI - Hypothalamic obesity in a patient with craniopharyngioma: dysregulation of neurohormonal control of energy balance. PMID- 15800427 TI - Natural history of patients with lumbar disc herniation observed by magnetic resonance imaging for minimum 7 years. AB - OBJECTIVE: The aim of this work was to elucidate the relation between the clinical course and morphologic changes of lumbar disc herniation on magnetic resonance imaging (MRI). METHODS: Twenty-one patients with lumbar disc herniation treated nonsurgically were followed for a minimum of 7 years and investigated with regard to their clinical outcome and the initial, 2-year, and final stage MRI findings. The space-occupying ratio of herniation to the spinal canal and the degree of disc degeneration were evaluated on serial MRI. RESULTS: The mean space occupying ratio of herniation showed significant reduction both on the 2-year and on the final scans. Progression of degeneration of the intervertebral disc was seen in all patients at the final investigation. Comparing patients with and without symptoms, no factors were detected on the initial and 2-year MR images capable of distinguishing patients who were and were not destined to develop lumbago and/or sciatica in the future. Morphologic changes of lumbar disc herniation continued to occur even after 2 years. CONCLUSIONS: Clinical outcome did not depend on the size of herniation or the grade of degeneration of the intervertebral disc in the minimum 7-year follow-up. PMID- 15800428 TI - Lumbar synovial cysts of the spine: an evaluation of surgical outcome. AB - OBJECTIVE: Our aim was to study the outcomes and results of surgically treated patients with synovial cysts of the lumbar spine in our institution. METHODS: Retrospective data from 39 consecutive patients, treated during the period of December 1996 to August 2004, were analyzed. Twenty-eight men (70%) and 11 women (30%) of mean age 63.3 years were studied. All pre- and postoperative signs, symptoms, extension/flexion radiographs, magnetic resonance imaging (MRI), and computed tomography (CT) with or without myelography were reviewed. All underwent surgery for synovial cysts with excision and decompression. Additional fusion in 26 patients was performed; 22 of them had degenerative spinal spondylolisthesis. Nine (23%) patients had prior decompression procedures, with three (8%) having had prior spinal instrumentation. Surgical outcomes were evaluated according to a questionnaire scoring system (scale of 1-4; 4 = excellent, 3 = good, 2 = fair, 1 = poor). Various preoperative attributes such as gender, age, weight, and height were analyzed to see if they had any effect on the outcome of surgery. Modified musculoskeletal outcomes data evaluation and management system (MODEM), questionnaire was provided to all; 24 (62%) responded. The following categories were determined: excellent (<20), very good (21-40), good (41-60), fair (61-80), and poor (81-100). Postoperative complications were also recorded. RESULTS: All patients had pain in their lower extremities, with 62% experiencing pain bilaterally. Ninety-five percent had pain in their back and 36% in the buttocks (36%). Eighteen (46%) patients had CT myelography. A total of 42 cysts were found. Two patients had bilateral cysts at L4-L5 level. Histology revealed two hemorrhagic cysts. The average duration of surgery was 231 minutes (range 92-391 minutes), and a mean blood loss of 930 mL (range 200-2500 mL) was recorded. Two operative dural tears and one postoperative wound dehiscence were observed. One patient had a recurrent synovial cyst at the site of original surgery. Eight patients (four each in the fusion and nonfusion group) had junctional degeneration and symptoms. A regression analysis performed on age, height, weight, and gender showed that they were not determining factors of surgical outcome. Surgery of spinal cysts at L4-L5 segment produced good and those at L5 S1 and multilevel excellent results. Patients with spinal segment fusion had superior outcomes, with 80% having excellent or good outcomes versus approximately 70% without fusion. With the modified MODEM questionnaire, 22 of the 24 (92%) patients scored between excellent, very good, and good. Two patients scored in the fair range, and none of the 24 patients scored in the range of poor. CONCLUSIONS: Spinal cysts are commonly found at the L4-L5 level, the site of maximum instability. MRI is the tool of choice for diagnosis. The etiology is still unclear, but underlying spinal instability has a strong association for formation of spinal cysts and worsening symptoms. Synovial cysts resistant to conservative therapy should be treated surgically. Resection and decompression with fusion remain an appropriate option. The optimal approach for patients with juxtafacet cysts remains unclear. The best surgical treatment approach for each particular individual appears to remain speculative. PMID- 15800429 TI - Morphometric analysis of the working zone for endoscopic lumbar discectomy. AB - OBJECTIVE: Our study's purpose was to analyze the working zone for the current practice of endoscopic discectomy at the lateral exit zone of the intervertebral foramen (IVF) and to define a safe point for clinical practice. METHODS: One hundred eighty-six nerve roots of the lumbar IVFs of cadaveric spines were studied. Upon lateral inspection, we measured the distance from the nerve root to the most dorsolateral margin of the disc and to the lateral edge of the superior articular process of the vertebra below at the plane of the superior endplate of the vertebra below. The angle between the root and the plane of the disc was also measured. RESULTS: The results showed that the mean distance from the nerve root to the most dorsolateral margin of the disc was 3.4 +/- 2.7 mm (range 0.0-10.8 mm), the mean distance from the nerve root to the lateral edge of the superior articular process of the vertebra below was 11.6 +/- 4.6 mm (range 4.1-24.3 mm), and the mean angle between the nerve root and the plane of the disc was 79.1 degrees +/- 7.6 degrees (range 56.0-90.0 degrees ). CONCLUSIONS: The values of the base of the working zone have a wide distribution. Blind puncture of annulus by the working cannula or obturator may be dangerous. The safer procedure would be the direct viewing of the annulus by endoscopy before annulotomy; the working cannula should be inserted into the foramen as close as possible to the facet joint. PMID- 15800430 TI - Tectorial membrane injuries in children. AB - We report three cases of tectorial membrane injury in children. An increased interspinous ratio was identified on cervical spine radiographs. The tectorial membrane injuries were diagnosed by magnetic resonance imaging. The three children were restrained passengers in high-speed motor vehicle accidents, and all sustained polytrauma. Two children with partial tears of the tectorial membrane were immobilized in a halo, and one with a longitudinal tear of the tectorial membrane had an occiput-to-C2 fusion. PMID- 15800431 TI - Vertebral rotation and thoracic torsion in adolescent idiopathic scoliosis: what is the best radiographic correlate? AB - BACKGROUND: As a result of the increased appreciation of the three-dimensional nature of scoliosis and modern spinal instrumentation's improved corrective capabilities, there has been renewed interest in the correction and measurement of vertebral rotation. Computed tomography (CT), the gold standard for accuracy, is limited in its clinical utility owing to cost, radiation exposure, and the effects of postural changes on scoliosis curves and vertebral rotation. Consequently, the Perdriolle and Nash-Moe techniques remain the standard measurements for providing a reasonable estimate of pre- and postoperative vertebral rotation because of their simplicity. However, these techniques have considerable interobserver variability, and pedicle screw instrumentation obscures the landmarks necessary for utilizing these techniques for postoperative vertebral rotation assessment. The purpose of the present study was to assess the utility of alternate radiographic measures to assess vertebral rotation and thoracic torsion when compared with conventional measures on pre- and postoperative radiographs and CT evaluation. METHODS: We reviewed the preoperative, immediate postoperative, and final follow-up radiographs, as well as the pre- and postoperative CT scans, of 19 patients (average age 15 years, 6 months) with Lenke 1 curves (average 55 degrees , range 47-66 degrees ), all treated with anterior spinal fusion. Coronal and sagittal Cobb angles as well as vertebral rotation (Perdriolle and Nash-Moe) at the superior uninstrumented, superior instrumented, apical, inferior instrumented, and inferior uninstrumented vertebrae were measured on all films, and vertebral rotation was assessed on the CT scans by a previously described method. Additionally, several measures of thoracic torsion (as a proxy for vertebral rotation and overall deformity improvement) were assessed. These included the rib-vertebral angle difference (RVAD), apical rib hump prominence (RH), apical vertebral body-rib ratio (AVB-R), and apical rib spread difference (ARSD). RESULTS: The postoperative main thoracic curve averaged 26 degrees (range 16-39 degrees , 52% correction) and 29 degrees (range 22-40 degrees , 47% correction) at final follow-up. For apical derotation, the postoperative CT improved from -11.5 degrees to -6.6 degrees and correlated significantly with the Cobb main thoracic curves (42% correction, r = 0.48, P = 0.003). There was weakly positive, but statistically significant, correlation between the pre- and postoperative CT scans and the corresponding Perdriolle and Nash-Moe measures of segmental rotation (r = 0.32-0.40, all P < 0.0001). The RVAD demonstrated poor correlation with the main thoracic curve values and correction, Perdriolle rotation and correction, and CT rotation and correction (r = -0.22 0.37, all P > 0.20). The apical RH demonstrated good correlation with the main thoracic curve (r = 0.65, P < 0.0001), apical Perdriolle rotation (r = 0.57, P < 0.0001), and CT apical rotation (r = 0.53, P = 0.002). We also found moderate correlation between the AVB-R and the main thoracic Cobb, apical Perdriolle, and CT (r = 0.57, 0.59, and 0.49, respectively; all P < 0.005). Similar relationships were found with the ARSD (r = 0.51, 0.47, and 0.43, respectively; all P < 0.02). CONCLUSIONS: The RH, AVB-R, and the ARSD-measures of thoracic torsion demonstrated moderate to good overall correlation with the main thoracic curve Cobb angles, apical Perdriolle rotation, and apical CT rotation. These should be useful as clinical measures for assessing three-dimensional deformity correction on plane radiographs, especially for the intraoperative evaluation of vertebral derotation and thoracic symmetry restoration. PMID- 15800432 TI - Residual neck pain after traumatic spondylolisthesis of the axis. AB - OBJECTIVE: Most patients with traumatic spondylolisthesis of the axis are treated nonsurgically. Some patients do not develop symptoms, but others experience strong and persistent neck pain. To clarify the factor that plays a major role in residual neck pain after this fracture, we reviewed nine patients who underwent conservative treatment. METHODS: Patients were assessed via telephone interviews an average of 62 months after injury and were divided into two groups: those with and those without neck pain. Angulation and anterior translation between the second and third cervical vertebrae were measured on lateral radiograms at the time of injury, removal of brace, and final follow-up. Fracture lines were also investigated by computed tomography. RESULTS: At the time of final follow-up, the average angulation was 6.3 degrees and the average translation was 5.3 mm in the neck-pain group and 0.6 degrees and 0.2 mm, respectively, in the no-symptom group. In patients with fracture lines present on the articular surface of the inferior face joints, angulation persisted or increased, even though the fractures were hemilateral. This was compatible with severe neck pain. In contrast, angulation and translation were improved in patients with an intact inferior articular surface of the axis, and pain was not reported. CONCLUSIONS: Injury on the inferior articular surface of the axis may disturb spontaneous healing of C2-C3 subluxation and cause residual neck pain. It is necessary to assess the presence of injury to the inferior facet bilaterally with plain or computed tomograms. PMID- 15800433 TI - Reliability analysis for digital adolescent idiopathic scoliosis measurements. AB - OBJECTIVE: Analysis of adolescent idiopathic scoliosis (AIS) requires a thorough clinical and radiographic evaluation to completely assess the three-dimensional deformity. Recently, these radiographic parameters have been analyzed for reliability and reproducibility following manual measurements; however, most of these parameters have not been analyzed with regard to digital measurements. The purpose of this study is to determine the intra- and interobserver reliability of common scoliosis radiographic parameters using a digital software measurement program. METHODS: Thirty sets of preoperative (posteroanterior [PA], lateral, and side-bending [SB]) and postoperative (PA and lateral) radiographs were analyzed by three independent observers on two separate occasions using a software measurement program (PhDx, Albuquerque, NM). Coronal measures included main thoracic (MT) and thoracolumbar-lumbar (TL/L) Cobb, SB MT Cobb, MT and TL/L apical vertical translation (AVT), C7 to center sacral vertical line (CSVL), T1 tilt, LIV tilt, disk below lowest instrumented vertebra (LIV), coronal balance, and Risser, whereas sagittal measures included T2-T5, T5-T12, T2-T12, T10-L2, T12 S1, and sagittal balance. Analysis of variance for repeated measures or Cohen three-way kappa correlation coefficient analysis was performed as appropriate to calculate the intra- and interobserver reliability for each parameter. RESULTS: The majority of the radiographic parameters assessed demonstrated good or excellent intra- and interobserver reliability. The relationship of the LIV to the CSVL (intraobserver kappaa = 0.48-0.78, fair to excellent; interobserver kappaa = 0.34-0.41, fair to poor), interobserver measurement of AVT (rho = 0.49 0.73, low to good), Risser grade (intraobserver rho = 0.41-0.97, low to excellent; interobserver rho = 0.60-0.70, fair to good), intraobserver measurement of the angulation of the disk inferior to the LIV (rho = 0.53-0.88, fair to good), apical Nash-Moe vertebral rotation (intraobserver rho = 0.50-0.85, fair to good; interobserver rho = 0.53-0.59, fair), and especially regional thoracic kyphosis from T2 to T5 (intraobserver rho = 0.22-0.65, poor to fair; interobserver rho = 0.33-0.47, low) demonstrated lesser reliability. In general, preoperative measures demonstrated greater reliability than postoperative measures, and coronal angular measures were more reliable than sagittal measures. CONCLUSIONS: Most common radiographic parameters for AIS assessment demonstrated good or excellent reliability for digital measurement and can be recommended for routine clinical and academic use. Preoperative assessments and coronal measures may be more reliable than postoperative and sagittal measurements. The reliability of digital measurements will be increasingly important as digital radiographic viewing becomes commonplace. PMID- 15800434 TI - Limited clinical utility of pain drawing in assessing patients with low back pain. AB - OBJECTIVE: The aim of this study was to assess the use of pain drawing by studying its ability to identify patients with low back pain and abnormal psychological profile. The intraevaluator repeatability of the penalty point method of scoring of pain drawing was also evaluated. METHODS: A total of 331 consecutive patients with low back pain were prospectively recruited. The psychologic profile was assessed using the Hospital Anxiety and Depression Scale. The pain drawings were scored with the penalty point method. The ability of pain drawing to identify patients with significant anxiety and depression was assessed by calculating the sensitivity, specificity, and positive predictive value. The intraevaluator repeatability was calculated for scoring done at an interval of 1 month using the kappa statistic. RESULTS: There were statistically significant differences in the anxiety and depression scores in patients with normal (n = 200) and abnormal (n = 131) pain drawing (anxiety: 9 +/- 4 vs 10.3 +/- 3.7; depression: 8.1 +/- 3.5 vs 8.8 +/- 3.6; P < 0.005). The pain drawing had a low sensitivity for detecting patients with any degree of anxiety (43%) or depression (40%). The positive predictive value of pain drawing for anxiety and depression was 78% and 69%, respectively. The kappa value for intraobserver assessment was 0.6 (P < 0.05). CONCLUSION: Though there are differences in anxiety and depression scores in patients with normal and abnormal pain drawing, the performance characteristics of pain drawing are less than acceptable and therefore limit its use in clinical practice. PMID- 15800435 TI - Material sensitivity study on lumbar motion segment (L2-L3) under sagittal plane loadings using probabilistic method. AB - OBJECTIVE: In this study, the probabilistic responses of a three-dimensional finite element L2-L3 motion segment, with and without posterior elements, tested under sagittal plane loadings, are presented. Understanding the effect of biologic uncertainties and variations on the biomechanical response provides an insight into spinal behavior under normal and degenerated conditions. METHODS: The biologic variability of 19 spinal components (nucleus, annulus, ligament, cortical/cancellous bone, endplate, and ligaments) in the motion segment was incorporated using statistical distributions into the model. A total of 2000 runs were performed using Monte Carlo probabilistic algorithms to compute the probabilistic response. RESULTS: This study establishes the relative importance of the spinal components in resisting the loading modes. The results show that for an intact motion segment, posterior ligaments are more dominant than intervertebral disc in resisting flexion moment. In extension, the capsular ligaments were found to be the most influential parameter. The intervertebral disc (ie, nucleus and annulus) affects the angular response of the disc body segment more than the hard tissues (ie, cortical and cancellous bone). CONCLUSIONS: The application of the probabilistic analysis provides a new approach whereby the influences of inherent uncertainties and variations in biologic structures can be studied and the biomechanical response assessed. PMID- 15800437 TI - Anterior correction of cervical spine lordosis using an adjustable depth tap: technical note. AB - OBJECTIVE: In cases of multilevel anterior cervical spine decompression where intervening vertebral bodies need to be incorporated into the anterior plate construct, it may be difficult to apply the plate flush against the entire anterior cortical surface of the spine segment. Some have suggested using the levering property of bicortical screws to "pull" the body against the plate. Others have recommended contouring the anterior cervical plate with a plate bender. The objective of this work was to describe a novel technique using the adjustable depth tap provided in the Atlantis plating system for achieving cervical spine alignment against the lordotic plate after multilevel anterior decompression. METHODS: We describe an illustrative case of a 14-year-old girl with complex cervical spine fractures after trauma. We used the 4.0-mm adjustable depth tap in the Atlantis plating system to lever the C4 body against the premachined lordotic plate. RESULTS: Cervical lordosis and anterior cervical plate placement were accomplished after multilevel anterior cervical decompression with the use of an adjustable depth tap. CONCLUSIONS: Bicortical screw placement and plate bending have drawbacks in trying to apply the anterior cervical plate firmly to the anterior surface of the cervical spine. We describe a new and safe technique of using an adjustable depth tap for achieving that same goal without any obvious drawbacks. PMID- 15800436 TI - Microsurgical posterior herniotomy with en bloc laminoplasty: alternative method for treating cervical disc herniation. AB - OBJECTIVE: At the present time, the anterior cervical discectomy and fusion procedure is widely accepted for treating cervical disc herniation. Recently, however, several authors have reported new disease due to degeneration of an adjacent segment. On the other hand, posterior discectomy, which can preserve mobility at the affected disc level, has been considered risky and technically difficult, especially for central or paracentral disc herniation. We are performing a new surgical technique, microsurgical posterior herniotomy with en bloc laminoplasty, for patients with myelopathy and radiculomyelopathy caused by cervical disc herniation. METHODS: Here, the surgical outcomes and radiographic changes were retrospectively investigated. Thirty patients (13 patients with myelopathy, 13 patients with radiculomyelopathy, and 4 patients with C5 dissociated motor loss) who underwent this procedure were reviewed. The average age was 50 years (range 31-70 years), and the average follow-up period was 28 months (range 12-76 months). Neurologic improvements were evaluated using the Japanese Orthopaedic Association (JOA) Scoring System as well as radicular pain and deltoid muscle power. Postoperative axial symptoms were scored, and radiographic changes were noted. RESULTS: The mean JOA score improvement was 74.2% (range 27.3-100%). In all 13 patients, preoperative radicular pain completely resolved after surgery. Deltoid power (in cases of C5 dissociated motor loss) markedly increased postoperatively. Cervical lordosis significantly increased at the time of the last follow-up (P = 0.01). The postoperative axial symptom score significantly correlated with the numbers of opened laminae (P = 0.03). CONCLUSIONS: This technique was safe and effective. Radiographically, the range of motion in the cervical spine and at the affected disc levels was preserved. In the future, this surgical procedure can become an alternative method for cervical disc herniation treatment. PMID- 15800438 TI - Diagnostic efficacy of thin slice CT in osteoid osteoma of the thoracic spine: report of two cases. AB - We present a 24-year-old man and a 31-year-old woman who complained of persistent back pain with osteoid osteoma of the thoracic spine. Computed tomography (CT) revealed a round sclerotic lesion in the posterior element of the thoracic spine, although their plain radiographs showed no abnormalities except a slight scoliosis. The patients underwent total excision of the tumor via a posterior approach. They are currently asymptomatic with no recurrence of the lesion and have returned to full activity. The thin slice CT is one of the most important diagnostic tools for osteoid osteoma of the spine. PMID- 15800439 TI - Intradural migration of broken IDET catheter causing a radiculopathy. AB - This case report describes the migration of a broken intradiscal electrothermal therapy catheter tip from the disc space into the thecal sac, leading to a radiculopathy that improved after removing the catheter. PMID- 15800440 TI - Discography-induced acute lumbar disc herniation: a report of five cases. AB - The reported complication rate of provocative lumbar discography is low, ranging from 0-2.5%. We report five cases of acute lumbar disc herniation precipitated by discography, a previously unreported complication. The cases reported comprise of four men and one woman with ages ranging from 23-45 years. All developed an acute exacerbation of radicular leg pain following multilevel provocative lumbar discography. One patient developed an acute foot drop. Comparison of lumbar MRI scans before and after discography demonstrated either a new herniated disc fragment or an increase in size of a preexisting herniation in all cases. On review of each discogram study and pre-discogram MRI an annular tear or small disc herniation was noted in all cases. In each case the patients' symptoms failed to resolve necessitating surgical intervention in all cases. In conclusion, annular deficiency is an obvious predisposing factor to discogram related disc herniation. New onset or a persistent exacerbation of radicular symptoms following provocative discography merits further investigation. PMID- 15800441 TI - Atypical presentation of sinus histiocytosis with massive lymphadenopathy as an epidural spinal cord tumor: a case presentation and literature review. AB - Sinus histiocytosis with massive lymphadenopathy is a benign lymphoproliferative disorder growing in frequency as awareness of the disease increases. Also known as Rosai-Dorfman disease, it typically presents as painless cervical lymphadenopathy with fever and malaise. A review of the literature reveals approximately 400 cases of extranodal involvement and approximately 44 cases of central nervous system involvement. Less than 10 of the reported central nervous system tumors have presented as an epidural spinal cord tumor. The authors describe the case of a 29-year-old woman with progressive paraplegia and leg pain. Magnetic resonance imaging of the thoracic spine revealed a tumor mass from T5 to T9. The tumor was resected, and the spinal column was stabilized with pedicle screw fixation and fusion. Postoperative treatment was initiated with radiation and physical rehabilitation. The following is a case report with literature review of the entity. PMID- 15800442 TI - Tuberculoma of the cervical spinal canal mimicking en plaque meningioma. AB - A previously healthy, HIV-negative, 40-year-old man presented with a 2-month history of progressive weakness of his left arm. Magnetic resonance imaging revealed an intradural, extramedullary plaque-shaped lesion at C6-T1 levels with high contrast enhancement. Based on the patient's clinical and radiologic findings, it was believed that the patient had an en plaque meningioma, and he was operated on. Histologic examination of the mass revealed granulomas with multinucleated and Langhans-type giant cells, typical of a tuberculoma. Intradural extramedullary tuberculomas should be considered in the differential diagnosis of en plaque meningioma as a rare entity. PMID- 15800443 TI - Pelvic ring reconstruction with a vascularized pedicle iliac bone graft for a large sacral schwannoma. AB - A case of pelvic ring reconstruction with a vascularized pedicle iliac bone graft for removal of a huge sacral schwannoma is reported. It is necessary to reconstruct a pelvic ring when its integrity is impaired. Spinal instrumentation is essential for a temporary stability of the pelvic ring in the course of complete healing, which at times results in instrumentation failure. Several cases of pelvic ring reconstruction using vascularized fibular bone grafts have also indicated positive results attributable to their good blood circulation, but techniques with them are relatively complicated. We applied a vascularized pedicle iliac bone graft to the pelvic ring reconstruction after resection of a huge sacral schwannoma. When a stable pelvic ring was regained by using a lumbosacral instrumentation technique, a tricortical iliac bone graft with its vascular pedicle was harvested, transported into the dead space, and tied to the right residual sacrum and the left ilium. The vascularized graft healed rapidly, and continuity of the pelvic ring was regained. This method is effective for pelvic ring reconstruction in that it does not require microvascular techniques and rapid bone healing can be obtained. PMID- 15800444 TI - Congenital absence of a lumbar pedicle presenting with contralateral lumbar radiculopathy. AB - The authors describe a rare case of a congenital absence of the right L5 pedicle in a 54-year-old man presenting with low back pain and radicular pain of his left leg. Plain x-ray films, computed tomography scan (CT) after myelography, and three-dimensional CT revealed the absence of the L5 pedicle and anomaly of the L4 L5 facet joint on his right side. On the left side, there were severe degenerative changes that were thought to be the result of an overload and instability. The degenerative changes led to late-onset neurologic impairment of the contralateral side, which was treated with spinal fusion. To our knowledge, this is the first report of contralateral symptoms due to unilateral defect of the facet joint accompanied by aplasia of a pedicle. PMID- 15800445 TI - Letter to the editor. PMID- 15800446 TI - Recurrent pruritic papules in the elbows of a 23-year-old man. PMID- 15800447 TI - [New therapeutic approaches in glioblastomas]. AB - INTRODUCTION: Current treatment of glioblastomas relies on surgical resection, radiotherapy and chemotherapy. However, the efficacy of these therapeutics is still limited and new therapeutic approaches based on the understanding of brain tumor biology are emerging. STATE OF ART: High expression of the EGF receptor by tumor cells, activation of the PI3K/Akt and the Ras/Raf pathways represent interesting targets for new selective drugs under development. Proteases inhibitors and antiangiogenic agents are also under investigations in clinical trials. Perspectives. In addition, the recent development of convection-enhanced delivery technique allows the administration of drugs which do not cross the blood-brain-barrier, such as selective toxins or immunostimulating oligonucleotides. CONCLUSION: Though the results of clinical trials have been somewhat disappointing, using different drug combinations or drug-radiotherapy associations will probably enable an improvement in the prognosis for these patients in the future. PMID- 15800448 TI - [Polyneuropathy associated with neurofibromatosis]. AB - INTRODUCTION: We review the literature on the spectrum of polyneuropathies associated with neurofibromatosis 1 (NF1) and 2 (NF2). BACKGROUND: Symptomatic neuropathies in NF1 are rarer than in NF2, but constitute a potentially severe complication associated with frequent morbidity related to the neuropathy itself, spinal cord compression or peripheral nerve sheath tumor malignant degeneration. Neuropathies are typically observed in young men with subcutaneous neurofibromas (NF1) or cutaneous schwannomas (NF2) and are characterized by a chronic slowly worsening sensorimotor polyneuropathy of the lower limbs. In NF1, demyelinating neuropathy may occur alone or in association with axonal features, whereas in NF2, axonal neuropathies are reported. Large multinodular roots and nerves, which can be easily detected by limb MRI, are characteristic features of NF1-associated polyneuropathies. PERSPECTIVES AND CONCLUSION: Although an associated pathogenic factor may reveal an asymptomatic neuropathy, patients should be monitored carefully because of the increased morbidity and mortality related to the significant proportion of malignant nerve-sheath tumors in these patients with NF1. PMID- 15800449 TI - [Creatine deficiency syndromes]. AB - INTRODUCTION: Creatine deficiency syndromes are a newly described group of inborn errors of metabolism affecting creatine metabolism. Three diseases have been described: deficiency of arginine: glycine amidinotransferase (AGAT), deficiency of guanidinoacetate methyltransferase (GAMT) and creatine transporter defect (CRTR). STATE OF ART: These syndromes are characterized by a depletion of creatine/phosphocreatine in the brain. Clinically, most of the patients develop a variable mental retardation and a severe speech delay associated with epilepsy, extra-pyramidal syndrome and behavior disturbances. These diseases are often diagnosed during infancy but a few adult cases have been reported recently. Diagnosis is established by measurement of guanidinoacetate and creatine in biologic fluids and in vivo proton magnetic resonance spectroscopy by the total lack of intra-cerebral creatine/phosphocreatine demonstrating. GAMT and AGAT deficiencies are treatable by oral creatine supplementation whereas patients with CRTR do not respond to the treatment. CONCLUSION: Better knowledge of these syndromes is necessary to optimize diagnosis and patient management of these rare but potentially treatable disorders. PMID- 15800450 TI - [A global intervention program for institutionalized demented patients]. AB - INTRODUCTION: Over the last decades many approaches have been developed to manage cognitive and behavioral disturbances in dementia. The present work describes a global intervention program carried out with moderately to severely demented institutionalized patients. The aims of the intervention program are to stimulate and maintain the preserved abilities of demented patients in a supportive context, to decrease the behavioral disturbance and to avoid burnout of care-unit staff. METHODS: This intervention combines different means: psychosocial care (validation therapy, social interaction), cognitive stimulation (memory and verbal training), and motor and sensitive stimulation. The global intervention program requires a special trained team composed of a supervisor, six aid-nurses, an occupational therapist, a speech therapist, a psychomotor therapist and a psychologist. The team cared for the patients five days per week over a three month period. Assessments were conducted before and after the intervention program to measure the benefit. RESULTS: Positive effects were shown for cognitive abilities, nutritional problems and staff burnout. However, due to the small sample size for this study, more research is needed to verify the effectiveness of this global intervention program, particularly the implications for nutrition. CONCLUSION: This global intervention combined with pharmacological treatment seems to be useful for managing psychological and behavioral disorders of institutionalized demented patients. PMID- 15800451 TI - [Specific remedial therapy in a specialist unit: evaluation of 31 children with severe, specific language or reading disorders over one academic year]. AB - INTRODUCTION: Up to 3 percent of the children in France present severe and specific language and/or reading disorders, despite regular remedial therapies. Few studies have measured the effectiveness of treatment administered in a specialist unit. PATIENTS AND METHOD: The aims of this study, focusing on children diagnosed as dysphasic and/or dyslexic, were: During the academic year 2001-2002, 31 children (18 dyslexic and 13 dysphasic) were attending school in our unit. The teaching program and intensive speech therapy (3 hours/week) were tailored for each child according to his/her specific disorders. Reading, spelling and numeracy developmental skills of each child were evaluated by appropriate tools at the beginning and at the end of the year. Impairment was defined by measuring the gap between the observed and the expected skills, according to each child's age. Using a self-control method, progress achieved by each child throughout the year was calculated with each tool, in each subject, by subtracting the impairments disclosed at the beginning from those disclosed at the end of the year. Progression was classified within three groups according to the progress normally expected over an academic year (i.e. nine months) from children with no disabilities attending school regularly; a progression fewer than three months was considered as no progression. Uni- and multivariate analyses including age (< or= or />9), type of pathology (dysphasia/dyslexia), and intellectual quotient (IQ) as covariates was carried out to search for independent prognosticators. RESULTS: The entire group demonstrated during the year significant progress for reading (p = 0.0001), spelling (p = 0.0001) and numeracy (p = 0.0001). Nineteen children (61 percent) showed more progress in reading than normally expected over nine months. Out of the remaining 12 children, 10 demonstrated more progress in spelling and/or numeracy than normally expected over nine months. All three reading evaluation tools disclosed a progression although one was less efficient (p = 0.05). Multivariate analysis disclosed age< or=9 and dysphasia as independent progress prognosticators. CONCLUSION: Placement in a specialist unit allows children suffering from severe dyslexia and dysphasia to lessen the gap in reading, spelling and numeracy. The two prognosticators disclosed highlight the importance of early diagnosis (i.e. before nine years old) and treatment of specific language and/or reading disorders. PMID- 15800452 TI - [Epidemiology and management of stroke patients in emergency departments of the Centre region of France]. AB - INTRODUCTION: Stroke is a leading cause of death and disability. Patients with suspected stroke are usually managed in emergency departments (ED). Stroke units must be created in our country, but only few French epidemiological data are available to define needs in stroke care. OBJECTIVE: A prospective study was planned to evaluate epidemiology and stroke care for patients with suspected stroke admitted in the 22 ED of our region in the center of France during a two month period. METHODS: Patients with suspected stroke seen at the 22 ED were prospectively followed until discharge or one month after admission. Data on demographic characteristics, mode of transport, delay of arrival and imaging, clinical findings at arrival, department of admission, diagnosis, Rankin scale at day 7, in-hospital mortality, and outcome at a month were collected. A global statistical analysis and a comparison between rural and urban ED were performed. RESULTS: Five hundred and ninety-eight patients were included. Mean age was 75 years. Median admission delay was 4 hours and 52 minutes. Predominant mode of transport was a private ambulance. A CT scan was obtained in 91 p.cent of cases with a median delay of 2 hours and 30 minutes. Only a third of the patients were admitted in departments of Neurology. Final diagnosis was: ischemic stroke (61 p.cent), transient ischemic attack (16 p.cent), hemorrhagic stroke (10 p.cent), other vascular disease (3 p.cent), non vascular disease (10 p.cent). In-hospital mortality was 20 p.cent, factors significantly associated with death rate were elevated age and a Glasgow coma scale<10 at admission. Mean length of stay was 12 days for stroke patients. At one month, 63 percent of patients were discharged to their home, 28 percent were transferred to an institution or in a rehabilitation unit, and 15 percent were still hospitalized. Significant differences in stroke care were found between rural and urban ED. CONCLUSION: This prospective study provides epidemiological data for our region. Creation of stroke units and definition of acute stroke networks are necessary to improve stroke care. PMID- 15800453 TI - [Presentation of Niemann-Pick type C disease with psychiatric disturbance in an adult]. AB - INTRODUCTION: Niemann-Pick Type C disease (NPC) is an autosomal recessive neurovisceral lysosomal lipid storage disorder. CASE REPORT: A 31-year-old right handed woman had suffered from schizophrenia for 13 years. At 25 years of age, she developed a gait disorder with a static and kinetic cerebellar syndrome, dysarthria, vertical supranuclear gaze palsy and cognitive impairment. Brain MRI was normal. Abdominal ultrasonography was performed because of hypercholesterolemia and elevated transaminases and revealed hepatosplenomegaly, which in conjunction with other signs and symptoms, suggested the diagnosis of NPC. The diagnosis was confirmed by demonstration of lysosomal storage of unesterified cholesterol (filipin staining) and of a reduced rate of LDL-induced cholesterol esterification. Implication of the NPC1 gene was assessed by genetic complementation analysis. DISCUSSION: The phenotypic presentation of NPC is remarkably variable. The rarer adult-onset form has a slowly progressive course. Psychotic manifestations are often prominent and may precede neurologic symptoms. Exposure to neuroleptics delays the diagnosis of NPC. CONCLUSION: Psychotic manifestations associated with cerebellar syndrome, vertical supranuclear gaze palsy, and splenomegaly are very suggestive of NPC disease which can be reliably diagnosed on cultured skin fibroblasts by filipin staining. PMID- 15800454 TI - [Cerebral angiitis caused by periarteritis nodosa]. AB - INTRODUCTION: We report a case of periarteritis nodosa revealed by a cerebral angiitis which recovered under treatment. OBSERVATION: A 52-year-old patient suddenly presented with a left sensory syndrome and a fluctuating aphasia due to ischemia involving both parietal lobes. The diagnosis of periarteritis nodosa was based on the following criteria: severe loss of weight, renal insufficiency, hypertension, angiography suggesting an arteritis. Instead of an ileo-cecal perforation, the patient recovered under corticosteroid and immunosuppressive therapy. CONCLUSIONS: Stroke in periarteritis nodosa may occur early be and associated with a good outcome. PMID- 15800455 TI - [Progressive BK virus associated multifocal leukoencephalopathy in an immunocompromised patient treated with corticosteroids]. AB - INTRODUCTION: BK virus has been described previously in renal transplant patients, but has also been reported in cases of progressive multifocal leukoencephalopathy. METHOD: A case of progressive BK virus multifocal leukoencephalopathy induced by long-term corticosteroid therapy is reported. RESULTS: In our case, BK virus was detected in cerebrospinal fluid by polymerase chain reaction (PCR). A detailed review of similar cases in the literature showed most of them occurred in AIDS patients, but four involved immunocompetent patients. CONCLUSION: Though BK virus infection usually leads to urinary tract symptoms in immunologically suppressed patients, it is important to be aware of its neurological manifestations and to recognize BK infections that can occur in immunocompetent patients. PMID- 15800456 TI - [Mutation of the aprataxin gene presenting with Charcot-Marie-Tooth-like neuropathy and cerebellar ataxia]. AB - BACKGROUND: Phenotype-genotype correlations, generally based on predominant associated signs, are being increasingly used to distinguish different types of autosomal recessive cerebellar ataxias (ARCA). CASE REPORTS: Two brothers developed signs of cerebellar ataxia with peripheral axonal motor and sensory neuropathy, distal muscular atrophy, pes cavus and steppage gait as seen in Charcot-Marie-Tooth neuropathy. The examination also showed oculomotor apraxia. Sural nerve biopsy revealed conspicuous reduction in the density of myelinated fibres but preservation of unmyelinated nerve fibres. Blood tests revealed low serum albumin and elevated cholesterol. A homozygous W279X truncating mutation was identified in exon 6 of the APTX gene, confirming the diagnosis of cerebellar ataxia with oculomotor apraxia type 1 (AOA1). CONCLUSIONS: These cases illustrate the presentation of AOA1 type of ARCA and discuss the role of peripheral neuropathy in the differential diagnostic of the ARCAs variants. PMID- 15800459 TI - [Hereditary Creutzfeldt-Jakob disease caused by a mutation at codon 200]. AB - INTRODUCTION: A typical case of genetic Creutzfeldt-Jakob disease in a 39-year old woman without remarkable familial history is described. CASE REPORT: Initial symptoms were disequilibrium, cerebellar syndrome and complex neurovisual complaints. EEG was pseudoperiodic. NSE and 14-3-3 protein levels were elevated in the CSF. MRI showed anomalies of the anterior parts of the putamen and the caudate nuclei on the MRI T2 FLAIR sequence, mainly on diffusion sequences. A quinacrine test did not yield any effect. Death eventually occurred 8 months after the first symptoms. CONCLUSION: Current data on genetic Creutzfeldt-Jakob disease are briefly reviewed. PMID- 15800460 TI - [Open and closed schizencephalie]. PMID- 15800461 TI - [Behavioral and psychological symptoms in dementia: description and management]. AB - Behavioral and Psychological Symptoms in Dementia (BPSD) are, beside cognitive disorders, major features of Alzheimer's disease and related disorders. Diagnosis is important to enhance our knowledge of the pathophysiology of dementia and of their functional consequences for patients and caregivers. Pharmacological and non-pharmacological management of dementia depends to a large extent on the presence of BPSD. A committee of geriatricians, neurologists and psychiatrists specialized in dementia (THEMA 2) has promoted an epidemiological, diagnostic and therapeutic update in this field. This work was based on the BPSD Consensus Conference Report edited in 2000 by the International Psychogeriatric Association. This report was updated with the most recent literature reports, and was adapted to the French environment. This paper is a synthesis of this meeting, validated and corrected by the entire Thema 2 group. PMID- 15800462 TI - Oral ulcers in children under chemotherapy: clinical characteristics and their relation with Herpes Simplex Virus type 1 and Candida albicans. AB - OBJECTIVE: The objective of this study was to determine the clinical characteristics of oral ulcers in pediatric oncology patients undergoing chemotherapy and their relation with the presence of Herpes Simplex Virus (HSV) type 1 and Candida albicans. STUDY DESIGN: The sample consisted of 20 ulcerative lesions from 15 children treated with chemotherapy in the Pediatric Service of the Regional Hospital of Concepcion, Chile. Two calibrated clinicians performed clinical diagnosis of the ulcers and registered general data from the patients (age, general diagnosis, absolute neutrophil count, and number of days after chemotherapy) and clinical characteristic of the ulcers: number, size, location, presence or absence of pain and inflammatory halo, edge characteristics, and exudate type. Additional to clinical diagnosis, culture for Candida albicans (C) and polymerase chain reaction (PCR) for Herpes Simplex Virus type 1 was performed. RESULTS: Ten ulcers occurred in patients with acute lymphoblastic leukemia, five in patients with acute myeloblastic leukemia and five in patients with other neoplastic diseases. Eight ulcers were HSV (+) / C (-), 6 HSV (-) / C (-), 4 HSV (+) / C (+) and 2 HSV (-) / C (+). Preferential location was the hard palate. Most lesions were multiple, painful, with inflammatory halo, irregular edges and fibrinous exudate. The average size was 6,5 millimeters, and the mean number of days after chemotherapy was 7.5 days. CONCLUSIONS: Oral ulcers in children with oncological diseases did not present a specific clinical pattern. They were strongly associated with HSV. PMID- 15800463 TI - Ki-67 expression in cytologic scrapes from oral squamous cell carcinoma before and after 24 gray radiotherapy--a study on 43 patients. AB - OBJECTIVE: To study the Ki-67 labeling indices in surface scrape smears from patients with oral squamous cell carcinoma before and after 24 Gray radiotherapy. STUDY DESIGN: Forty three patients with histologically documented squamous cell carcinoma of the oral cavity were sampled by means of surface scrape smears prior to therapy and after receiving 24 Gray fractionated radiotherapy. These smears were stained for Ki-67 expression using the avidin biotin alkaline phosphatase technique. RESULTS: Ki-67 expression was seen in an extremely small number of cells. Only 10 tumors showed positive cells, and the labeling index in them varied from 0.1 % to 0.01 %. After 24 Gray irradiation, no case showed any Ki-67 positive cells. CONCLUSIONS: The overall yield of malignant cells in surface smears is low even prior to therapy and their number decreases further after irradiation. This, along with other factors including low concentration of proliferating cells on the surface of the lesion and obscuring inflammatory cells, anucleate squames, bacterial colonies and proteinaceous material could have accounted for the low labeling indices obtained. Radiation induced decline in proliferation has been described previously. The major conclusion, in balance, is that conventional oral scrape cytology may not be the optimal tool for immunocytochemical evaluation of proliferation in oral squamous cell cancer. PMID- 15800464 TI - Temporomandibular disorders: referred cranio-cervico-facial clinic. AB - The bond between temporomandibular disorders and referred craniofacial symptomatology is more and more evident. In it subsists the prevailing necessity of understanding the temporomandibular disorders and the cranio-cervico-facial referred symptomatology from a neurophysiologic and muscle-skeletal perspective contained in the stomatognatic system. Diagnosis in head and neck areas is difficult because of its complex anatomy. Some painful craniofacial syndromes exhibit the same symptoms although they don.t seem objectively possible and that is what confuses the specialist and the patient. Pain in the head and the neck is one of the most complex to diagnose because of its varied origins that can be neurological, vascular, muscular, ligamental and bony. This article seeks to show some reasonable anatomical and pathophysiological connections of this muscle skeletal disorder expressed with symptoms like tinnitus, otic fullness, otalgia and migraine among others. Disciplines in health such as neurology, the otolaryngology and dentistry share common anatomical and pathophysiological roads constructed in an increased muscular activity that generates muscle-skeletal disorders and is difficult to locate referred craniofacial symptomatology. This revision aspires to sensitize the medical specialist and the odontologist in the understanding of the important interdisciplinary handling in the detection of this disorder. This offers better tools in the conservative therapy phase of this craniofacial referred symptomatology. PMID- 15800465 TI - Candida albicans, Staphylococcus aureus and Streptococcus mutans colonization in patients wearing dental prosthesis. AB - BACKGROUND: Denture stomatitis is associated to Candida albicans, different bacteria and other co-factors such as an acid pH, a carbohydrate ingestion increase, different systemic illnesses and pharmacological treatments. OBJECTIVE: The aim of this study was to determine Candida albicans, Staphylococcus aureus and Streptococcus mutans prevalence in the mucous membrane and prosthesis of patients with and without atrophic denture stomatitis and its relationship with other potential clinical co-factors. STUDY DESIGN: Saliva was collected from 105 patients (62 female and 43 male) wearing dental prosthesis in order to measure their pH. Oral samples of the mucous membrane and the internal surface of dental prosthesis were taken with sterile cotton to proceed with the microbiological study. The identification of the isolated microorganisms was performed using conventional microbiological methods. RESULTS: Diabetes and Hypertension were the most frequent systemic illnesses. High carbohydrate ingestion was observed in numerous patients. Atrophic denture stomatitis was reported in 50 patients and the pH average in saliva was of 5.2. The presence of C albicans, S. aureus and S. mutans in the mucous membrane and prosthesis was of 51.4%, 52.4% and 67.6%, respectively. C. albicans was isolated in 66.7% from the prosthesis, whereas S. aureus and S. mutans were isolated in 49.5% of those same prosthesis. C. albicans was isolated in 86% of the patients with atrophic denture stomatitis and S. aureus was isolated in a similar percentage (84% of patients). The isolation of S. mutans was less frequent, and it was observed in 16% of the oral samples of these patients. CONCLUSIONS: C. albicans, S. aureus and S. mutans frequently colonize the oral mucous of patients wearing dental prosthesis. This illness bearing condition is more frequent in patients with denture stomatitis, even though dental prosthesis colonization is lower than in the oral mucous. PMID- 15800466 TI - Functions of the cytokines in relation oral lichen planus-hepatitis C. AB - BACKGROUND: Lichen planus (LP) is a chronic common mucocutaneous inflammatory disorder of uncertain aetiology. An association between hepatitis C virus (HCV) infection and LP has been recognised, particularly in Italy, Spain and Japan. The pathogenesis of such an association is unclear, but it may be due to cell mediated cytotoxicity to an epitope shared by HCV and damaged keratinocytes. Recent studies using in situ hybridization suggest that HCV may replicate in the oral mucosa. OBJECTIVES: The aim of the present study was to examine the oral epithelium of patients with oral LP for evidence of HCV-RNA by polymerase chain reaction (PCR) and to examine the relationship to cytokines including interferon (INF-gamma), interleukins (IL-1, IL-2, IL-4, IL-6, IL-8 , and IL-10), tumour necrosis factor (TNF-alpha) and transforming growth factor (TGFbeta-1). PATIENTS/METHODS: We selected 100 Italian patients, and divided them into 4 groups. Group A consisted of 25 HCV+ve patients with erosive oral LP. Group B was a control group constituted by 25 healthy HCV -ve subjects with no LP. Group C consisted of 25 HCV-ve patients with oral reticular LP and Group D was made of 25 HCV-ve patients with oral erosive LP. The patients of group A (test group) were submitted to oral biopsy with 2 samples of epithelium, lesional and non-lesional, and a 10 ml peripheral blood sample was taken. The patients of group B (negative control), C and D (comparison groups) were submitted to oral epithelial biopsy and a 10 ml peripheral blood sample was collected. PCR was used to search for HCV RNA in biopsy material. Cytokines INF-gamma ,IL-1, IL-2, IL-4, IL-6, IL-8 , IL-10 and TNF-alpha and TGFbeta-1 were assayed in serum. RESULTS AND CONCLUSIONS: PCR did not detect the viral genome in oral epithelium of the patients with oral LP and HCV+ve (group A), but there was an increase in levels of TNF-alpha and a reduction of IL-1, INF-gamma and IL-8 compared to patients who had oral reticular LP but HCV-ve and to patients who had oral erosive LP but HCV-ve, and compared to negative controls. The results indicate that patients of group A showed a reduction of pro-inflammatory but an increase in immunomodulant cytokines. The results suggest the possibility that HCV exerts an indirect effect, mediated possibly by the induction of cytokines and lymphokines. PMID- 15800467 TI - Implant rehabilitation of the atrophic upper jaw: a review of the literature since 1999. AB - The severely resorbed maxilla presents serious limitations for conventional implant placement. As a result, different techniques have been developed in the last two decades, with variable results. The most significant approaches comprise the placement of implants in anatomical abutments, elevation of the sinus floor, and reconstructive surgery with bone grafting. The present study reviews the most important articles on the management of the atrophic upper jaw published in the literature since 1999. PMID- 15800468 TI - Basal cell nevus syndrome. Presentation of six cases and literature review. AB - Basal cell nevus syndrome, also known as Gorlin-Goltz syndrome, is an autosomal dominant inherited disorder which is characterised by the presence of multiple maxillary keratocysts and facial basal cell carcinomas, along with other less frequent clinical characteristics such us musculo-skeletal disturbances (costal and vertebrae malformations), characteristic facies, neurological (calcification of the cerebral falx, schizophrenia, learning difficulties), skin (cysts, lipomas, fibromas), sight, hormonal, etc. On occasions it can be associated with aggressive basal cell carcinomas and malignant neoplasias, for which early diagnosis and treatment is essential, as well as family detection and genetic counselling. Currently there are new lines of investigation based on biomolecular studies, which aim at identifying the molecules responsible for these cysts and thus allowing an early diagnosis of these patients. In its clinical management and follow up, the odonto-stomatologist, the maxillofacial surgeon and several other medical specialists are involved. In this paper a review of the literature, and six cases of patients affected by multi-systemic and varied clinical expression of basal cell nevus syndrome, are presented. PMID- 15800469 TI - Periapical surgery using the ultrasound technique and silver amalgam retrograde filling. A study of 71 teeth with 100 canals. AB - INTRODUCTION: Periapical surgery using ultrasound allows the treatment of root canals of difficult access, with the sacrifice of little root tissue. As a result, periapical disorders which were condemned to treatment failure in the past can now be dealt with successfully. MATERIAL AND METHODS: In 71 teeth presenting 100 root canals treated with ultrasound and subjected to retrograde filling with silver amalgam, the course and short-term success of management was evaluated in relation to lesion size, the magnitude of apical resection, and the size of the retrograde filling cavity. The duration of follow-up was one year, with post-treatment controls after 6 and 12 months. RESULTS: After 6 months, the percentage clinical and radiological success was 92% and 58%, respectively. One year after periapical surgery the corresponding percentages were 95% and 80%. Global success after 6 months was 63%, versus 84.2% after 12 months. No statistically significant relation was observed between treatment success and the size of the periapical lesion, the amount of apex resected, or the size of retrograde filling. CONCLUSION: Periapical surgery using ultrasound and retrograde filling with silver amalgam affords a high success rate after 12 months. PMID- 15800470 TI - Undecalcified bone samples: a description of the technique and its utility based on the literature. AB - The evaluation of bone regeneration and peri-implantary bone apposition requires laboratory techniques that allow the evaluation of hard tissues without needing to decalcify them. In this study, we describe the cutting-grinding procedure (EXAKT system) for obtaining samples of undecalcified bone and aim to establish the procedures for their histological and histomorphometric evaluation. A review is made of the literature on histological and histomorphometric evaluation using undecalcified samples in surgical and oral implantology procedures in which bone regeneration or bone apposition is produced on the dental implants. The technique is a lengthy process, and is principally indicated in the investigation of bone biology. The use of undecalcified samples allows the differentiation of immature from mature bone, and the quantification of variables such as the bone-implant interface, the density of the bone area and the speed of bone apposition. PMID- 15800471 TI - DNA cytometry of oral leukoplakia and oral lichen planus. AB - BACKGROUND: DNA cytometry is a technique that allows densitometric analysis of nuclear DNA of cells and in this way the evaluation of DNA ploidy. OBJECTIVE: The purpose of the present study was the evaluation of DNA ploidy in oral leukoplakia and oral lichen planus and correlation with histological dysplasia. METHODS: We analyzed oral incisional biopsies from a group of 40 patients with oral leukoplakias (20 homogeneous and 20 non-homogeneous) and 45 patients with oral lichen planus (25 erosive, 20 reticular), by DNA cytomorphometry, after conventional histological examination of sections stained with haematoxylin and eosin. RESULTS AND CONCLUSIONS: Eight of 20 non-homogeneous leukoplakias showed aneuploidy; (3 showed mild aneuploidy) and only 2 specimens of erosive lichen planus showed moderate aneuploidy, while all 20 cases of reticular lichen planus were diploid. There was not concordance between DNA ploidy and the degree of histological assessed dysplasia. PMID- 15800472 TI - Improving influenza, pneumococcal polysaccharide, and hepatitis B vaccination coverage among adults aged <65 years at high risk: a report on recommendations of the Task Force on Community Preventive Services. AB - The Task Force on Community Preventive Services conducted systematic reviews to evaluate the effectiveness of interventions to improve targeted vaccination coverage (i.e., coverage with vaccines recommended for some but not all persons in an age range on the basis of risk for exposure or disease) among adults aged <65 years at high risk when implemented alone (single-component interventions) and in combination with other interventions (multicomponent interventions). A 1999 report by the Task Force examined the effectiveness of interventions to increase coverage with universally recommended vaccinations (i.e., vaccines recommended for all persons in particular age groups). Three targeted vaccinations recommended for populations at risk are addressed in this review: influenza, pneumococcal polysaccharide, and hepatitis B. The Task Force identified evidence that certain combinations of interventions have improved vaccination coverage. To increase targeted vaccination coverage, the Task Force recommends a combination of interventions that include selected interventions from two or three categories of interventions (i.e., increasing community demand for vaccinations, enhancing access to vaccination services, and provider- or system-based interventions). The Task Force also recommends provider reminders, when implemented alone, to improve targeted vaccination coverage. This report provides additional information about population-based interventions to improve the coverage of influenza, pneumococcal polysaccharide, and hepatitis B vaccines among populations at risk, briefly describes how the reviews were conducted, and provides information that can help in applying the interventions locally. PMID- 15800473 TI - Tobacco use, access, and exposure to tobacco in media among middle and high school students--United States, 2004. AB - Two of the national health objectives for 2010 are to reduce the prevalence of any tobacco use during the preceding month to < or =21% and the prevalence of current cigarette use to < or =16% among high school students (objectives 27-2a and 27-2b). The National Youth Tobacco Survey (NYTS), conducted by CDC in 2004, provided estimates of current use of tobacco products and selected indicators related to tobacco use, including youth exposure to tobacco-related media and access to cigarettes. This report summarizes data from the 2004 NYTS and describes changes in tobacco use and indicators related to tobacco use since 2002. During 2002-2004, middle school students reported decreases in pipe use, seeing actors using tobacco on television or in movies, and seeing advertisements for tobacco products on the Internet. Among high school students, no changes were observed in the use of tobacco or in access to tobacco products; however, seeing actors using tobacco on television or in movies declined slightly, and seeing advertisements for tobacco products on the Internet increased. The lack of substantial decreases in the use of almost all tobacco products among middle and high school students underscores the need to fully implement evidence-based strategies (e.g., increasing the retail price of tobacco products, implementing smoking-prevention media campaigns, and decreasing minors' access as part of comprehensive tobacco-control programs) that are effective in preventing youth tobacco use. PMID- 15800474 TI - Inadvertent laboratory exposure to Bacillus anthracis--California, 2004. AB - On June 9, 2004, the California Department of Health Services (CDHS) was notified of possible inadvertent exposure to Bacillus anthracis spores at Children's Hospital Oakland Research Institute (CHORI), where workers were evaluating the immune response of mice to B. anthracis. This report summarizes the subsequent investigation by CDHS and CDC, including assessment of exposures, administration of postexposure chemoprophylaxis, and serologic testing of potentially exposed workers. The findings underscore the importance of using appropriate biosafety practices and performing adequate sterility testing when working with material believed to contain inactivated B. anthracis organisms. PMID- 15800475 TI - Estimated influenza vaccination coverage among adults and children--United States, September 1, 2004-January 31, 2005. AB - In response to the unexpected shortfall in the 2004--05 influenza vaccine supply, CDC recommended in October 2004 that vaccine be reserved for persons in certain priority groups, including persons aged > or =65 years and 6--23 months, persons aged 2-64 years with conditions that increased their risk for influenza complications, residents of chronic-care facilities, close contacts of infants aged <6 months, and health-care workers with direct patient contact. In late December 2004, based on declining demand among these groups, two additional groups (i.e., healthy persons aged 50-64 years and household contacts of all persons at high risk) were added to the list of vaccination priority groups. To monitor influenza vaccination coverage during the 2004-05 season, the Behavioral Risk Factor Surveillance System (BRFSS), an ongoing, state-based, telephone survey of civilian, noninstitutionalized persons, added new questions to collect information on priority status and the month and year of vaccination for adults and children. This report is based on analysis of data collected during February 1-27, 2005, regarding respondent-reported receipt of influenza vaccination during September 1, 2004-January 31, 2005. The results of this analysis indicated that influenza vaccination coverage levels through January 2005 among adults in priority groups nearly reached those in recent years, whereas coverage levels among adults not in priority groups were approximately half of levels in 2003, in part because 9.3% of those unvaccinated persons in nonpriority groups declined vaccination this season. The results further suggested that designation of the priority groups successfully directed the nation's influenza vaccine supply to those at highest risk. In addition, vaccination coverage among children aged 6-23 months was notable (48.4%), given that 2004-05 was the first year this group was recommended for influenza vaccination . PMID- 15800476 TI - Influenza vaccine prebooking and distribution strategies for the 2005-06 influenza season. AB - For the 2004-05 influenza season, CDC, in coordination with the Advisory Committee on Immunization Practices (ACIP), issued interim influenza vaccine use recommendations after Chiron Corporation announced that none of its inactivated influenza vaccine (Fluvirin) would be available in the United States. To plan for the upcoming 2005-06 influenza season, CDC has met with influenza vaccine manufacturers, including those intending to apply for approval to sell in the United States, to develop supply projections and distribution strategies, including prebooking (i.e., advance ordering of vaccine) and partial shipment of orders to those customers who prebook. As of March 25, 2005, the supply of inactivated influenza vaccine projected for the 2005-06 season appeared adequate to meet the historical demand from persons in the priority groups established by ACIP during the 2004-05 season. If more vaccine becomes available, additional groups can also be targeted for vaccination. PMID- 15800477 TI - Outbreak of Marburg virus hemorrhagic fever--Angola, October 1, 2004-March 29, 2005. AB - On March 23, 2005, the World Health Organization (WHO) confirmed Marburg virus (family Filoviridae, which includes Ebola virus) as the causative agent of an outbreak of viral hemorrhagic fever (VHF) in Uige Province in northern Angola. Testing conducted by CDC's Special Pathogens Branch detected the presence of virus in nine of 12 clinical specimens from patients who died during the outbreak. PMID- 15800483 TI - New insight into the pathogenesis of sepsis and the sepsis syndrome. PMID- 15800484 TI - Prognostic factors of surgical resection in middle and distal bile duct cancer: an analysis of 55 patients concerning the significance of ductal and radial margins. AB - BACKGROUND: The surgical outcome of middle and/or distal bile duct cancer remains unsatisfactory. Although the resectional margin is known to be a predictive factor, the prognostic significance of a positive ductal margin and other radial margin has never been evaluated independently. METHODS: The clinicopathologic data of 55 patients who had undergone surgical resection for middle and/or distal bile duct cancer between 1987 and 2003 were reviewed retrospectively. The surgical procedures consisted of pancreatoduodenectomy in 42 patients (76%), extrahepatic bile duct resection in 8 patients (15%), major hemihepatectomy (Hx) in 3 patients (5%), and pancreatoduodenectomy plus Hx in 2 patients (4%). In all the patients, intraoperative diagnosis of the ductal margins was performed using frozen sections. Twenty-one clinicopathologic factors, including the status of the ductal margins and of other radial margins, were evaluated using univariate and multivariate analyses. RESULTS: The overall 5-year survival rate and the median survival time were 24% and 38 months, respectively. There were 4 (7%) postoperative deaths. Fifteen of the remaining 51 patients (29%) were determined to have positive hepatic-side ductal margins during operation, and 14 of them underwent additional resection of the bile duct (1.6[range, 1-3] times, on average). As a result, hepatic-side ductal margin (hm) and duodenal-side ductal margin were found to be positive in 6 and 0 patients on the final pathologic analysis, respectively. Two of the 6 patients (33%) with positive hm have developed ductal recurrence so far, but the status of hm was not found to be a significant predictor. The depth of neoplastic invasion into the bile duct wall, pancreatic invasion, radial margin, and blood transfusion were significant prognostic factors by the univariate analysis. Multivariate analysis revealed that the depth of neoplastic invasion and blood transfusion were the independent prognostic factors. CONCLUSIONS: In the treatment of middle and distal bile duct cancer, it is of importance to secure a negative radial margin, although it may be less beneficial to obtain a negative hm. Surgeons should make efforts to obtain negative radial margins and to avoid blood transfusion. PMID- 15800485 TI - Hepatectomy for hepatocellular carcinoma with major portal or hepatic vein invasion: results of a multicenter study. AB - BACKGROUND: The role of hepatic resection in patients with hepatocellular carcinoma (HCC) and invasion of a main portal or hepatic vein branch is controversial. We evaluated the efficacy of hepatic resection and the factors affecting survival after resection in such patients. METHODS: The records of 102 patients who underwent resection for HCC with major vascular invasion between 1984 and 1999 were reviewed. Prognostic factors were evaluated by univariate and multivariate analysis. RESULTS: The study included 87 men and 15 women. The median age was 59 years. The perioperative mortality rate was 5.9%. Median survival was 11 months (median follow-up, 93 months). The 1-, 3-, and 5-year survival rates were 45%, 17%, and 10%; the longest-living survivor was still alive at 14.8 years. Absence of moderate to severe fibrosis and absence of high nuclear grade were associated with a better 5-year survival rate (23% vs 5%; P = .001 and 21% vs 9%; P = .04, respectively). On multivariate analysis, moderate to severe fibrosis remained a significant predictor of both short-term (< or = 6 months) and long-term (>6 months) survival ( P < .03 and P < .01, respectively). CONCLUSIONS: Hepatic resection for HCC with major vascular invasion is associated with median survival exceeding historical survival in patients not treated surgically. Patients with HCC and major vascular invasion who derive long-term benefit from resection have no or minimal underlying fibrosis. PMID- 15800487 TI - Surgical treatment of peritoneal carcinomatosis from well-differentiated digestive endocrine carcinomas. AB - BACKGROUND: The presence of peritoneal carcinomatosis (PC) in association with endocrine carcinomas (EC) is generally considered to have no impact on life expectancy, contrary to liver metastases. This study was aimed at assessing the actual prognostic impact of PC and to evaluate a new treatment with respect to survival times. PATIENTS AND METHODS: Among 111 patients undergoing surgery for progressive, well-differentiated EC, 37 (33%) presented a histologically proven PC, with synchronous liver metastases in 36 of them. The origin was ileal or appendiceal (carcinoid tumors) in at least 81% of cases. The patients were divided into 2 groups. Patients in group 1 (n = 20) could not undergo complete resection of PC, while those in group 2 (n = 17) underwent complete cytoreductive surgery, followed by immediate intraperitoneal chemotherapy. Partial hepatectomy was performed in 65% of patients in group 2. The median follow-up was 6.9 years. RESULTS: There was no postoperative mortality, and the morbidity rate was 47%. In group 1, 15 of the 20 patients died (5-year survival rate, 40.9%). Deaths were caused either by liver failure (60% of patients) or bowel obstruction from PC (40%). In group 2, six of the 17 patients died (5-year survival rate, 66.2%; P = .007). These patients died of liver failure (n = 4, 23.5%), bowel obstruction (n = 1, 5.8%), and cerebral hemorrhage (n = 1, 5.8%). CONCLUSIONS: PC associated with EC is not a rare event; it is mainly caused by carcinoid tumors and is always associated with liver metastases. When present, PC is the direct cause of death in 40% of patients if no specific treatment is undertaken. Treatment of PC with maximal cytoreductive surgery and immediate intraperitoneal chemotherapy appears promising, even though it can only be considered as palliative. PMID- 15800488 TI - Invited commentary: Surgery and chemotherapy for peritoneal carcinomatosis--more about selection and biology than treatment? PMID- 15800489 TI - Reelevation of parathyroid hormone level after parathyroidectomy in patients with primary hyperparathyroidism: importance of decreased renal parathyroid hormone sensitivity. AB - BACKGROUND: We hypothesized that impaired peripheral sensitivity to parathyroid hormone (PTH) may play a role in reelevation of PTH after successful operation for primary hyperparathyroidism (pHPT). METHODS: Factors affecting reelevation of PTH were determined in 90 patients who underwent parathyroidectomy for pHPT. PTH/nephrogenous cyclic adenosine monophosphate ratio, as an index of renal resistance to PTH, was examined in relation to factors shown to influence reelevation of PTH. RESULTS: Serum PTH levels were elevated above the upper limit of normal in 23 patients (26%) at 1 week and in 39 patients (43%) at 1 month after parathyroidectomy. These 39 normocalcemic patients with elevated serum PTH at 1 month after parathyroidectomy had a higher preoperative serum level of PTH and lower serum phosphate and 25-hydroxyvitamin D (25OHD) concentrations than those with normal PTH (n = 59). Elevated PTH and low 25OHD were shown by multivariate analysis to be significant predictors of reelevation of PTH. Renal resistance to PTH was higher in patients with vitamin D deficiency or renal insufficiency than in patients with normal serum vitamin D concentrations or normal renal function, and it increased according to increases in levels of PTH. CONCLUSIONS: The mechanism of PTH reelevation in patients with pHPT after successful parathyroidectomy appears to be renal resistance to PTH. PMID- 15800490 TI - Sleep disturbances before and after parathyroidectomy for secondary hyperparathyroidism. AB - BACKGROUND: The prevalence of insomnia in patients with symptomatic secondary hyperparathyroidism and the improvements of sleep disturbances after parathyroidectomy have not been previously reported. METHODS: Thirty-one patients who had undergone successful total parathyroidectomy and autotransplantation for symptomatic secondary hyperparathyroidism were enrolled in the study. The symptoms of skin itching, bone pain, and general weakness were recorded. Preoperatively, serum levels of calcium, phosphorus, alkaline phosphatase (Alk ptase), intact parathyroid hormone (iPTH), am melatonin (noon), and pm melatonin (midnight) were measured in association with a simple yes/no questionnaire of 5 items about sleep disturbances as described previously. The severity of insomnia was the sum of positive items of sleeping disturbances. The sleep hours per night and the habit of taking sleeping pills were also recorded. One week after surgery, serum levels of calcium, phosphorus, Alk-ptase, iPTH, am melatonin, and pm melatonin were measured again. Three months after surgery, symptoms of skin itching, bone pain, and general weakness were recorded, and serum levels of calcium, phosphorus, Alk-ptase and iPTH were measured in association with a yes/no questionnaire of sleep disturbances. The severity of insomnia, sleep hours per night, and the habit of taking sleeping pills were recorded again. RESULTS: One week after parathyroidectomy, serum levels of calcium, phosphorus, and iPTH decreased significantly; serum levels of Alk-ptase and am melatonin increased significantly; serum levels of pm melatonin did not change significantly. Three months after parathyroidectomy, symptoms of skin itching, bone pain, and general weakness decreased significantly; serum levels of calcium, phosphorus, Alk-ptase, and iPTH decreased significantly; sleeping disturbances and severity of insomnia improved significantly in association with longer sleep hours per night. A significant reduction of the habit of taking sleeping pills was also noted. Preoperatively, the severity of insomnia was correlated with skin itching, general weakness, and levels of iPTH. Postoperatively, the severity of insomnia was correlated with skin itching, general weakness, and bone pain. CONCLUSIONS: Preoperatively, a high prevalence of sleep disturbances (97%) was found in patients with symptomatic secondary hyperparathyroidism. We conclude that nocturnal melatonin levels do not change after parathyroidectomy; the improvements in sleep disturbance and the decreases in severity of insomnia are found 3 months after surgery in association with longer sleep hours per night. Decreases of symptoms such as skin itching, bone pain, and general weakness may be the reasons for the improvement in sleep and the decrease in insomnia. PMID- 15800491 TI - Quick PTH assay cannot predict incomplete parathyroidectomy in patients with renal hyperparathyroidism. AB - BACKGROUND: Contradictory reports on the value of intraoperative quick parathyroid hormone (PTH) monitoring in renal hyperparathyroidism have been published. METHODS: Thirty-five consecutive patients underwent total parathyroidectomy, central neck dissection, bilateral thymectomy, and immediate autotransplantation. PTH levels were measured by PTH assay at induction of anesthesia (baseline level) and in 5-minute intervals after excision of the last parathyroid gland. Parathyroidectomy was considered "total" in patients with PTH levels <10 pg/mL (subgroup 1), "subtotal" between 10 and 65 pg/mL (subgroup 2) and "insufficient" at >65 pg/mL (subgroup 3) within the first postoperative week. RESULTS: Fifteen minutes after excision of the last gland, PTH levels dropped to 19.4 +/- 15.7% (subgroup 1), 14.9 +/- 5.9% (subgroup 2), and 18 +/- 6.7% (subgroup 3) from baseline among 22 patients on hemodialysis, to 22.1 +/- 18.7% and 17.5% in 9 patients (subgroups 1 and 2) after successful kidney transplantation, and to 10.7% and 17.5% (subgroup 1) and 12.8% and 31.4% (subgroup 2) in 4 patients with reduced renal function after kidney transplantation. CONCLUSIONS: Currently available QPTH assays are not useful to predict insufficient resection of hyperfunctioning parathyroid tissue. PMID- 15800492 TI - Induction of IL-6 within the rodent intestinal muscularis after intestinal surgical stress. AB - BACKGROUND: Postoperative ileus is a poorly understood surgical problem characterized by leukocyte extravasation into the intestinal muscularis and suppression in muscle function. The study objective was to delineate a mechanistic inflammatory cascade initiated by intestinal manipulation. METHODS: ACI and Sprague-Dawley rats, and IL-6 +/+ and IL-6 -/- mice were subjected to intestinal manipulation. One group of rats received adhesion molecule-blocking antibodies (1A29 and WT.3) before intestinal manipulation. Interleukin-6 (IL-6) messenger RNA (mRNA) levels and electrophoretic mobility shift assay for signal transducers and activators of transcription (STAT) activation were measured in tissue extracts. IL-6 protein levels were assessed by immunohistochemistry and enzyme-linked immunosorbent assay. RESULTS: IL-6 mRNA from muscularis extracts demonstrated a significant induction after intestinal manipulation. No IL-6 induction was observed in mucosal extracts. Adhesion molecule blockade resulted in a marked decrease of cellular infiltration but did not change IL-6 mRNA expression in muscularis extracts. Resident macrophages in the muscularis stained for IL-6 by immunohistochemistry after intestinal manipulation. The isolated manipulated muscularis demonstrated a significant increase in IL-6 release. Electrophoretic mobility shift assay of manipulated muscularis showed a marked increase in IL-6-dependent Stat3 activation. CONCLUSIONS: This study demonstrates that manipulation of the small bowel during an abdominal operation initiates downstream induction, translation, release, and functional activity of IL-6 within the muscularis. PMID- 15800493 TI - Relationship between iNOS expression and aortic cell proliferation and apoptosis in an elastase-induced model of aorta aneurysm and the effect of 1400 W administration. AB - BACKGROUND: In the present study, we employed an elastase infusion-dependent abdominal aortic aneurysm (AAA) model to examine inducible nitric oxide synthase (iNOS) expression in relation to cellular proliferation and apoptosis in this pathologic condition. Furthermore, we employed N-(3 (aminomethyl)benzyl)acetamidine (1400 W), a previously shown selective iNOS inhibitor, to further explore this relationship. METHODS: Adult male Wistar rats were randomized into separate groups. Group A served as a control and received an intra-aortic saline infusion, while groups B, C, and D received an intra-aortic elastase infusion according to standard protocols. The animals in group C were administered postoperatively the highly selective iNOS inhibitor, 1400 W, while rats in group D received regularly the same compound preoperatively and postoperatively. The animals were killed at postoperative days 7 and 14. Aorta diameter and nitric oxide (NO), nitrite/nitrate, and MDA levels were measured. iNOS expression was assessed by immunohistochemistry and Western blot analysis, while Ki-67 immunohistochemistry and TUNEL assay were used to evaluate cellular proliferation and apoptosis, respectively. RESULTS: Increased iNOS and NO levels accompanied aneurysm development in groups B, C, and D, but these levels were significantly lower in groups C and D, compared with group B. Interestingly, very low but detectable levels of iNOS were found in the control group, indicating a basal constitutive level. Cell growth parameters were augmented in group B compared with group A. In contrast, groups C and D exhibited a significant decrease of the cellular growth parameters but did not attain normal values. CONCLUSIONS: iNOS-derived NO is associated with the cellular growth parameters of the vessel cells, predominantly smooth muscle cells. Selective iNOS blockage ameliorates the cellular remodeling in AAAs. PMID- 15800495 TI - MMP-12 has a role in abdominal aortic aneurysms in mice. AB - BACKGROUND: Matrix metalloproteinase (MMP)-12 levels are increased in the abdominal aortic aneurysm (AAA), implicating this protease in AAA pathogenesis. The purpose of this study was to assess the role of MMP-12 in aneurysm formation. METHODS: A murine aneurysm model was generated by periaortic application of 0.25 mol/L calcium chloride (CaCl 2 ) for 15 minutes. Aortic diameters were measured and compared before and 10 weeks after aneurysm induction. Aortic diameter changes for wild type (WT) and MMP-12 knockout (MMP-12 -/- ) mice were determined. MMP-12 production in mouse aorta was analyzed by casein zymography. MMP-2 and MMP-9 expressions were examined by gelatin zymography. Immunohistochemical study was used to measure macrophage infiltration into the aorta. RESULTS: There is an increase of 63 +/- 5% (mean +/- SEM) in aortic diameters of WT mice after CaCl 2 inductions, while MMP-12 -/- mice increased only 26 +/- 14%. Connective tissue staining of aortic sections from WT mice showed disruption and fragmentation of medial elastic fibers, while MMP-12 -/- mice showed only focal elastic lamellae breakdown. MMP-12 levels in WT mice were significantly increased after CaCl 2 treatment, whereas no MMP-12 was detected in MMP-12 -/- mice. There was no difference in the MMP-2 and MMP-9 productions between WT and MMP-12 -/- mice. Immunohistochemical analysis demonstrated that infiltrating macrophages in the aorta of MMP-12 -/- mice were significantly less than WT controls. CONCLUSIONS: MMP-12 deficiency attenuates aneurysm growth, possibly by decreasing macrophage recruitment. PMID- 15800496 TI - Progressive fascial wound failure impairs subsequent abdominal wall repairs: a new animal model of incisional hernia formation. AB - BACKGROUND: Fascial wound failure alters the phenotype of the abdominal wall. This study introduces a novel animal model of progressive failure of the ventral abdominal wall fascia, which generates large incisional hernias. MATERIAL AND METHODS: A mechanistic model of incisional hernia was compared with a model of acute myofascial defect hernia repair. Using biological tissue repair markers, tensiometric measurements and recurrent hernia rate, we measured the mechanism by which incisional hernias regenerate abdominal wall structure and function after mesh and suture herniorrhaphy. RESULTS: Recurrent incisional hernia formation was significantly increased after repairs of the hernia model, compared with the myofascial defect model (6/16 vs 0/16, P < .05). In the hernia model, there were significant decreases in the recovery of wound strength, energy, and extensibility before mechanical disruption, compared with the myofascial defect model. Unexpectedly, excision of fascial hernia wound edges did not significantly improve tissue repair outcomes in the hernia model group. CONCLUSIONS: Clinically accurate animal modeling can recreate the wound pathology expressed in mature incisional hernias. Progressive fascial wound failure decreases the fidelity of subsequent incisional hernia repair, compared with identically sized acute abdominal wall defect repairs. The mechanism appears to include decreased fascial wound strength and decreased tissue compliance after herniorrhaphy. PMID- 15800497 TI - Invited commentary: The whistle-blower act (qui tam). PMID- 15800498 TI - Huge cystic communicating duplication of the right colon with perforated appendicitis. PMID- 15800499 TI - Solid pseudopapillary tumor of the pancreas diagnosed postpartum. PMID- 15800500 TI - Protein C deficiency as a cause of simultaneous acute thrombosis of the superior mesenteric vein and inferior vena cava with jejunal infarction. PMID- 15800504 TI - Status of activator methods chiropractic technique, theory, and practice. AB - OBJECTIVE: To provide an historical overview, description, synthesis, and critique of the Activator Adjusting Instrument (AAI) and Activator Methods Chiropractic Technique of clinical assessment. METHODS: Online resources were searched including Index to Chiropractic Literature, EBSCO Online, MANTIS, CHIROLARS, CINAHL, eJournals, Ovid, MDConsult, Lane Catalog, SU Catalog, and Pubmed. Relevant peer-reviewed studies, commentaries, and reviews were selected. Studies fell into 2 major content areas: instrument adjusting and the analysis system for therapy application. Studies were categorized by research content type: biomechanical, neurophysiological, and clinical. Each study was reviewed in terms of contribution to knowledge and critiqued with regard to quality. DISCUSSION: More than 100 studies related to the AAI and the technique were found, including studies on the instrument's mechanical effects, and a few studies on clinical efficacy. With regard to the analysis, there is evidence for good reliability on prone leg-length assessment, but to date, there is only 1 study evaluating the Activator Methods Chiropractic Technique analysis. CONCLUSION: A body of basic science and clinical research has been generated on the AAI since its first peer-reviewed publication in 1986. The Activator analysis may be a clinically useful tool, but its ultimate scientific validation requires testing using sophisticated research models in the areas of neurophysiology, biomechanics, and statistical analysis. PMID- 15800505 TI - On the subject of human subjects. PMID- 15800506 TI - Chiropractic in North America: a descriptive analysis. AB - OBJECTIVE: This paper provides descriptive data on chiropractors, their practice, and their patients in North America in the past decade. METHOD: Five sites in the United States and 1 in Canada were chosen, and a random sample of chiropractors was interviewed. In each practice, 10 patients were systematically selected on a single day. A total of 131 chiropractors and 1275 patients were interviewed. SUMMARY: The results suggest that doctors of chiropractic have firmly established themselves within the health care system in the United States and Canada and are able to attract patients who come to them directly for treatment, for largely back-related conditions, and who are willing to pay for their care. PMID- 15800507 TI - The Nordic back pain subpopulation program: a 1-year prospective multicenter study of outcomes of persistent low-back pain in chiropractic patients. AB - OBJECTIVES: The aims of the study are to describe the low-back pain and disability status at baseline, the fourth visit, and at 3 and 12 months in Norwegian patients treated by chiropractors for persistent low back pain (LBP) and to describe movements between various subgroups over time. DESIGN: Prospective uncontrolled multicenter study. METHODS: Self-reported pain was measured with a 0-10 box scale and disability with the revised Oswestry LBP questionnaire. The main outcome measures were mean pain or disability values and numbers of LBP-free patients. LBP status was assessed through patient questionnaires at baseline, the fourth visit, and after 3 and 12 months. STUDY SUBJECTS AND SETTING: Of 205 invited chiropractors, 115 Norwegian chiropractors were each willing to recruit 10 consecutive patients who had LBP for at least 2 weeks at the time of consultation and a minimum of 30 days altogether within the preceding year. The numbers of participants were 875 (baseline), 799 (fourth visit), 598 (3 months), and 512 (12 months). RESULTS: Considerable improvement was noted between baseline and the fourth visit both for mean values and in numbers of LBP-free patients. There was virtually no further mean improvement up to the third month, whereas the number of LBP-free individuals doubled. At 12 months, no additional improvement was noted, and 80% reported that they had experienced recurrent problems. Less than 1% reported considerable worsening. Severity of symptoms at baseline determined the subsequent outcome, mild symptoms tending to worsen, and severe symptoms tending to improve. CONCLUSION: The outcome pattern is similar to that found in other clinical studies. Treatment outcome should be measured early with follow-up at 3 rather than at 12 months, because patients will improve or recover quickly but may experience recurring problems. Numbers "cured" appear to be a feasible outcome variable in this type of study population. PMID- 15800508 TI - Recruiting seniors with chronic low back pain for a randomized controlled trial of a self-management program. AB - OBJECTIVE: To identify recruitment challenges and elucidate specific strategies that enabled recruitment of seniors for a randomized trial on low back pain comparing the Chronic Disease Self-management Program of the Stanford University to a 6-month wait-list control group. DESIGN: Recruitment for a randomized controlled trial. SETTING: Community-based program offered at 12 locations. PARTICIPANTS: Community-dwelling seniors 60 years and older with chronic low back pain of mechanical origin. METHODS: Passive recruitment strategies included advertisement in local and senior newspapers, in senior e-mail newsletters and listservs, in local community centers and businesses. Active strategies included meeting seniors at health fairs, lectures to the public and organizational meetings, and the help of trusted professionals in the community. RESULTS: A total of 100 white and 20 African American seniors were recruited. The program seemed to have the most appeal to white, middle-class older adults, educated through high school level. Advertisement failed to attract any participants to the program. Successful strategies included interaction with seniors at health fairs and lectures on health care, especially when the program was endorsed by a trusted community professional. CONCLUSION: Generating interest in the self management program required keen communication skills because the idea of "self management" was met with a myriad of responses, ranging from disinterest to disbelief. Generating interest also required active participation within the communities. Initial contacts had to be established with trusted professionals, whose endorsement enabled the project managers to present the concept of self management to the seniors. More complex recruitment strategies were required for this study involving the self-management approach to back pain than for studies involving treatment. PMID- 15800509 TI - Changes in cerebellar blood flow after manipulation of the cervical spine using Technetium 99m-ethyl cysteinate dimer. AB - BACKGROUND: Cervical spine manipulation is one of the many interventions practiced by health professionals to treat musculoskeletal disorders of the cervical spine. Although serious consequences of manipulation have been documented, the incidence is thought to be rare. More frequently, there may be minor transient side effects after manipulation of the cervical spine, such as headache, dizziness, and nausea. One of the hypothesis is that these side effects are caused by ischemia in the areas perfused by the vertebral arteries. OBJECTIVE: The purpose of this study was to investigate whether manipulation of the cervical spine can influence blood flow in the brain. METHODS: Single photon emission computed tomography was used to examine changes in regional cerebral blood flow caused by cervical spine manipulation (CSM) performed by a physiotherapist to 15 volunteers, using a 1-day split-dose Technetium 99m-ethyl cysteinate dimer single photon emission computed tomography activation paradigm. RESULTS: One brain region was identified showing a decreased regional cerebral blood flow after manipulation. This region was situated in the anterior lobe of the left cerebellum (-42, -48, -24). CONCLUSIONS: These findings suggest that cerebellar hypoperfusion may occur after CSM. This could explain why certain people experience headache, dizziness, or nausea after CSM. Further investigation into patient symptoms in the presence of cerebellar hypoperfusion and the possible link of these findings with other adverse reactions are warranted. PMID- 15800510 TI - Variance in manual treatment of nonspecific low back pain between orthomanual physicians, manual therapists, and chiropractors. AB - OBJECTIVE: The aim of the study was to identify differences in the diagnosis and treatment of nonspecific low back pain among 3 professional groups in the Netherlands: orthomanual physicians, manual therapists, and chiropractors. METHODS: Information was obtained from training materials from professional groups, literature searches, and observation of selected practitioners at work. RESULTS: In The Netherlands, there are differences in education between the 3 professional groups. The focus of orthomanual medicine is on abnormal positions of components of the skeleton and symmetry in the spine. Manual therapy focuses on functional disorders of the musculoskeletal system. Chiropractic focuses on the musculoskeletal and nervous systems in relation to patients' health in general. Orthomanual medicine considers inspection and palpation the most important diagnostic tools. Manual therapists and chiropractors additionally perform tests to determine functional disorders and manual therapists evaluate psychosocial influences. Chiropractors take radiographs if necessary. Orthomanual physicians apply mobilization techniques using fixed protocols. Manual therapists and chiropractors use various manipulation and mobilization techniques and their manipulation techniques differ in amplitude and velocity. CONCLUSIONS: Diagnostic techniques and treatment methods of the 3 professional groups differ considerably. For more accurate reporting of the efficacy of manipulative and mobilizing therapies, the characteristics of treatments should be described in more detail when reported in studies such as randomized clinical trials. PMID- 15800511 TI - Influence of axial rotation on chiropractic pelvic radiographic analysis. AB - OBJECTIVE: The aim of the study is to explore associations between axial (y-axis) rotation of the pelvis and pelvic radiographic measurements. STUDY DESIGN: Descriptive film statistics of an incrementally rotated pelvis and linear regression analysis were performed. METHODS: A phantom pelvic model was incrementally imaged (1 degrees increments) at 40-in source-to-image distance through 10 degrees of axial rotation. Chiropractic line drawing analysis was performed. The chiropractic examiner was blinded to the degree of rotation during the film analysis. Regression analysis was performed between axial rotation and pubic symphysis deviation, sacral width, and innominate and femur head heights. Each measurement corresponds to a chiropractic listing within the Gonstead system. RESULTS: Regression analysis revealed a strong association between the degree of axial rotation and each response variable: pubic symphysis deviation, sacral width, innominate height, and femur head height. The strongest relationship existed between y-axis rotation and pubic symphysis deviation. CONCLUSIONS: Chiropractic pelvic listings are strongly influenced by positioning of the subject. A few degrees of axial rotation may create apparent misalignments of several millimeters. PMID- 15800512 TI - Reliability of McKenzie classification of patients with cervical or lumbar pain. AB - BACKGROUND: In the McKenzie system, patients are classified first into syndromes, then into subsyndromes. At present, the reliability of classification with this system is unclear. No study has included patients with cervical pain, and the studies to date have reported conflicting results. OBJECTIVE: The aim of the study is to investigate the interexaminer reliability of the McKenzie classification system for patients with cervical or lumbar pain. SUBJECTS: Fifty patients with spinal pain (25 with lumbar pain and 25 with cervical pain) were included in the study. METHOD: The patients were assessed simultaneously by 2 physical therapists (14 in total) trained in the McKenzie method. Agreement was expressed using the multirater kappa coefficient and percent agreement for classification into (i) syndromes and (ii) subsyndromes. RESULTS: The reliability for syndrome classification was kappa = 0.84 with 96% agreement for the total patient pool, kappa = 1.0 with 100% agreement for lumbar patients, and kappa = 0.63 with 92% agreement for cervical patients. The reliability for subsyndrome classification was kappa = 0.87 with 90% agreement for the total patient pool, kappa = 0.89 with 92% agreement for lumbar patients, and kappa = 0.84 with 88% agreement for the cervical patients. CONCLUSION: The McKenzie assessment performed by persons trained in the McKenzie method may allow for reliable classification of patients with lumbar and cervical pain. PMID- 15800513 TI - Back belt use for prevention of occupational low back pain: a systematic review. AB - BACKGROUND: Back pain continues to be the leading overall cause of morbidity and lost productivity in the workplace. Recently, there has been a renewed interest in the use of back belts by industry to reduce occupational low back pain (LBP). OBJECTIVES: To examine the literature and evaluate the effectiveness of back belt use for the primary prevention of occupational LBP. METHODS: MEDLINE, CINAHL, EMBASE, and HEALTHSTAR were searched for relevant articles published up to July 2003. Studies were included if participants were material handlers, and outcomes included the incidence and/or duration of lost time of reported LBP among workers who wore back belts compared with those who did not. The quality of the evidence was scored independently by 2 reviewers using a double rating method, first according to research design followed by an internal validity rating. Final synthesis of the evidence was performed in which the evidence was classified as good, fair, conflicting, or insufficient. RESULTS: Ten epidemiologic studies meeting inclusion criteria were identified. Of 5 randomized controlled trials, 3 showed no positive results with back belt use; 2 cohort studies had conflicting results; and 2 nonrandomized controlled studies and 1 survey showed positive results. CONCLUSIONS: Currently, because of conflicting evidence and the absence of high-quality trials, there is no conclusive evidence to support back belt use to prevent or reduce lost time from occupational LBP. PMID- 15800514 TI - Chiropractic maintenance care and quality of life of a patient presenting with chronic low back pain. AB - OBJECTIVE: To report on a 26-year-old female patient presenting with uncomplicated chronic low back pain who received chiropractic maintenance care using 2 quality of life outcome assessment instruments. OUTCOME MEASURES: Short form (SF-36) subscales, Quality of Well-Being Scale, Visual Analog Scale, and number of tender vertebral spinous processes. RESULTS: After 9 months of care the SF-36 subscale scores showed improvement. The SF-36, although low before care, approached normal on 3 subscales and exceeded normal population values on 5 subscales after 9 months. The SF-36 physical and mental composite scores improved from mean baseline scores of 23.4 and 25.3 to 43.7 and 62.8, respectively, after 9 months of care. The Quality of Well-Being Scale scores improved from a mean pre intervention score of 1.1 to a post-intervention score of 8.2. The Visual Analog Scale scores improved from a mean pre-intervention score of 8 to a post intervention score of 1.5. The mean number of chiropractic vertebral subluxations, detected via palpation of spinous process tenderness, went from a pre-care mean of 6.5 to a post-care mean of 4. CONCLUSION: The patient appeared to experience improvement in quality of life while showing signs suggestive of improved spinal function. The relationship between indicators of vertebral subluxation and quality of life deserves further investigation using a research design that allows for exploration of possible causal relationships. PMID- 15800515 TI - Chiropractic treatment of lumbar spine synovial cysts: a report of two cases. AB - OBJECTIVE: To present the treatment of low back and radicular pain due to synovial cysts of the lumbar spine including chiropractic distraction manipulation and physiological therapeutic care. CLINICAL FEATURES: Two patients (71-year-old man and 59-year-old woman) with magnetic resonance imaging (MRI) diagnosed large synovial cysts at the L3 through L4 and L4 through L5 vertebral levels, respectively, had lower extremity pain, numbness, and tingling of the respective L4 and L5 dermatomes. INTERVENTION AND OUTCOME: Chiropractic distraction manipulation was performed at the level of the synovial cyst. The manipulation was performed daily until 50% pain relief was attained, followed by diminished frequency of care. Physiotherapy included positive galvanism, iontophoresis, tetanizing electrical stimulation, stabilization exercises, and home cryotherapy. The male patient's pain was reduced by 50% in 14 days and 100% at 60 days. Range of motion of the thoracolumbar spine increased, walking distance increased from 1 to 2 blocks to 1 mile without pain, and repeat MRI showed reduced size of the cyst. The female patient, under the same treatment regimen, was pain free in 6 weeks. CONCLUSION: Chiropractic distraction manipulation and physiological therapeutic care relieved 2 patients with low back and radicular pain attributed to MRI-confirmed synovial cysts of the lumbar spine. This treatment may be an initial conservative treatment option for synovial cysts with careful patient monitoring for progressive neurologic deficit which would necessitate surgery. Distraction manipulation may be a safe and effective conservative treatment of synovial cyst causing radicular pain; further data collection of clinical outcomes is warranted. PMID- 15800516 TI - Magnetic resonance imaging of the hip: detection of labral and chondral abnormalities using noncontrast imaging. AB - PURPOSE: Traditional imaging techniques have limited ability to detect subtle chondral and labral injuries of the hip. We performed a retrospective review of patients who underwent magnetic resonance imaging (MRI) of the hip and subsequent hip arthroscopy in order to evaluate the ability of optimized, noncontrast MRI to identify tears of the acetabular labrum and defects in articular cartilage. TYPE OF STUDY: Retrospective review of a consecutive sample. METHODS: Between January 1997 and July 2000, 92 patients had MRI of the hip, followed by arthroscopic surgery of that hip by 1 of 2 surgeons (R.B., D.E.P.). Two musculoskeletal MR radiologists blinded to the initial MRI and surgical findings, independently interpreted the studies, looking for the location and degree of articular cartilage and acetabular labral pathology. RESULTS: Of the 92 patients studied, each of 2 radiologists correctly identified 83 (94%) and 84 (95%) of the 88 labral tears present at surgery, respectively. There was 92% interobserver agreement on the MRI studies. For articular cartilage defects on the femoral head and acetabulum, there was good agreement (92% and 86% within 1 grade) between MRI and surgical grading and between the 2 MR readers (kappa of 0.8 for femoral head cartilage and 0.7 for acetabular cartilage). CONCLUSIONS: This study shows that noncontrast MRI of the hip, using an optimized protocol, can noninvasively identify labral and chondral pathology. Such information may facilitate deciding which patients warrant surgical intervention, thus preserving hip arthroscopy as a therapeutic tool. LEVEL OF EVIDENCE: Level II, Development of Diagnostic Criteria Study. PMID- 15800517 TI - Arthroscopic rotator cuff repair: the learning curve. AB - PURPOSE: The purpose of this study was to answer the question: How many cases are required for a surgeon to become proficient in performing arthroscopic rotator cuff repair? We hypothesize that as surgical experienced is gained, learning can be quantitatively shown by a significant decrease in operative time. TYPE OF STUDY: Prospective case series. METHODS: Rotator cuff repair time (RCRT) in minutes (as well as other time components comprising total surgical time) was recorded for 100 consecutive patients having arthroscopic rotator cuff repair performed by a single surgeon beginning with his first case in private practice. Mean RCRTs for consecutive blocks of 10 cases were compared. Learning is graphically represented by plotting the RCRT by case number and generating a logarithmic trend curve. A best-fit linear equation (y = mx + b) allows comparison of the initial 10 cases with the subsequent 90 cases, where m , the slope, represents the rate of decrease in RCRT (learning). RESULTS: Mean RCRT decreased significantly (P < .05) from the first block of 10 cases to the second block of 10 cases. There were no significant changes in mean RCRT when comparing other consecutive blocks of 10 cases. The slope of the line fitting the first block of 10 cases is -8.75; the slope (m) of the line fitting the subsequent 90 cases is -0.23. There is no significant difference in mean RCRT when cases are stratified by tear size. CONCLUSIONS: Graphic representation of RCRT by case number generates a learning curve whereby learning is quantitatively shown as a significant decrease in operative time as surgical experience is gained. CLINICAL RELEVANCE: Qualification of the learning curve for arthroscopic rotator cuff repair provides a guide for orthopaedic surgeons contemplating the expected time line for acquiring proficiency in this technique. PMID- 15800519 TI - Arthroscopic sliding knot: how many additional half-hitches are really needed? AB - PURPOSE: To evaluate the optimal number of additional half-hitches needed to achieve optimal knot-holding capacity (KHC) of lockable sliding knots. TYPE OF STUDY: Mechanical testing study. METHODS: Four configurations of arthroscopic knots (Duncan loop, Field knot, Giant knot, and SMC knot) were tested for their knot-holding capacity. For each knot configuration, 6 sequential knots were made including the initial sliding knot and an additional 5 knots by increasing the half-hitches 1 at a time. Each additional half-hitch was made as a reverse half hitch with alternate posts. For each sequential knot configuration, 12 knots were made using No. 2 braided sutures. On the Servo-hydraulic materials testing system (Instron 8511; MTS, Minneapolis, MN), cyclic loading, load to clinical failure (3 mm displacement), load to ultimate failure, and mode of failure were measured. RESULTS: Most of the initial loops without additional half-hitches showed dynamic failure with cyclic loading. However, after 1 additional half-hitch, all 3 (SMC, Field, and Giant) knots showed resistance to dynamic cyclic load. After 2 additional half-hitches, the Duncan loop was secured without slippage from the cyclic loading test. The mean displacement after the end of cyclic loading decreased with each additional half-hitch. In particular, the SMC and Giant knot reached plateau at 0.1-mm or less displacement after 1 additional half-hitch. The Field knot and Duncan loop needed 3 additional half-hitches. The SMC knot and Duncan loop needed 1 additional half-hitch to reach greater than 80 N at clinical failure, whereas the other 2 knots needed 2 additional half-hitches. For the load exceeding 100 N for clinical failure, the SMC knot required 3 additional half hitches and the other 3 knots needed 4 additional half-hitches. Addition of more than 3 half-hitches did not increase the load to clinical failure in the SMC knot. However, load to clinical failure increased up to 4 additional half-hitches in the other 3 knots (P < .05). The load to ultimate failure reached plateau when 3 or more additional half-hitches were made for all knot configurations. As the number of additional half-hitches increased, the mode of failure switched from pure loop failure (slippage) to material failure (breakage). The Duncan loop showed poor loop security--even with 5 additional half-hitches, some failed by slippage (17%). On the other hand, after 3 additional half-hitches, the 3 other knots showed greater than 75% of failure by material breakage mode (SMC and Field 92%, Giant 75%). CONCLUSIONS: Even with its own locking mechanism, a lockable sliding knot alone does not withstand the initial dynamic cyclic load. For all tested variables, the SMC knot required a minimum of 2 additional half-hitches. All knots showed a near plateau in knot security with 3 or more additional half hitches. The Duncan loop may need more than 3 additional half-hitches for optimal security. CLINICAL RELEVANCE: The study shows that the knots tested needed at least 2 additional half-hitches, so this should become standard clinical practice. PMID- 15800518 TI - Cartilage thickness of the talar dome. AB - PURPOSE: To perform a successful operation, accurate information on the articular cartilage thickness of the talus is essential. The purpose of this study was to determine the articular cartilage thickness of the talar dome in the area where osteochondritis dissecans is common. TYPE OF STUDY: Convenient samples. METHODS: Articular cartilage thickness of the mid talar dome was measured in 29 ankles of 17 cadavers. The average age of the specimens was 70 years (range, 53 to 91 years). Twenty-two were from men and 7 were from women. Coronal sections of the mid talar domes were obtained from the specimens measuring 2 mm in width and 5 mm in depth. Radiographs of the sectioned specimens were taken and were enlarged with a personal computer. The thickness of the articular cartilage was measured at 9 areas: medial gutter (area 1), medial corner (area 2), medial edge of the dome plateau (area 3), lateral edge of the dome plateau (area 7), lateral corner (area 8), lateral gutter (area 9), and quarter points between areas 3 and 7 (areas 4, 5, and 6). RESULTS: The average thickness of the total areas was 1.35 +/- 0.22 mm in the male specimens and 1.11 +/- 0.28 mm in the female specimens. The thickest was area 2, which represented the medial corner, and the thinnest was area 9, which represented the lateral gutter in both men and women. There was a relationship between the thickness of the cartilage and the width of the talar dome. CONCLUSIONS: The thickness was usually less than 2 mm. However, the cartilage thickness varies widely by gender, area, and individual. CLINICAL RELEVANCE: Our report will help surgeons predict the volume of cartilage that needs to be repaired and produce treatment at a reasonable cost without an excessive invasion of patients. PMID- 15800520 TI - Clinical evaluation of arthroscopically assisted anterior cruciate ligament reconstruction: patellar tendon versus gracilis and semitendinosus autograft. AB - PURPOSE: To evaluate the clinical results of anterior cruciate ligament (ACL) reconstruction in patients with ACL-deficient knee in 2 similar groups of patients. TYPE OF STUDY: A prospective randomized comparison of patellar tendon (PT) versus semitendinosus and gracilis tendon (STG) autografts for ACL reconstruction. METHODS: Between 1994 and 1996, 85 consecutive male patients with chronic ACL-deficient knees underwent arthroscopically assisted reconstruction with either autologous PT or double-loop STG (4-strand) graft. PT grafts were used in patients with even-numbered birth dates and STG grafts for those with odd numbered birth dates. Preoperatively, no significant differences between the 2 groups were noted with respect to age, level of activity, and degree of laxity (chi-square analysis). A standardized rehabilitation program was used for both groups postoperatively that included immediate active extension and early weight bearing and gradual flexion. Return to sports was permitted 8 months postoperatively. Assessment of the patients was carried out using a questionnaire, clinical assessment, Lysholm knee scores, the International Knee Documentation Committee scale, and radiological examination. RESULTS: At a mean follow-up of 81 months, there was no significant difference between the 2 groups with respect to subjective complaints (recurrent giving way, functional level) or objective laxity evaluation, including KT-1000 measurement or return to sports. Loss of extension of < or =5 degrees was greater in the PT group (12 patients, 30 %) than in the STG group (8 patients, 17 %). There was loss of flexion of < or =15 degrees in 5 patients (12 %) in the PT group and 1 patient (2.2 %) in the STG group. Anterior knee pain was recorded in 10 patients (24 %) in the PT group and 3 patients (5 %) in the STG group. The Lysholm knee score was 91.6 and 92.7 for the PT and STG groups, respectively, and the Tegner activity score decreased from 8.9 preoperatively for both groups to 7.9 for the PT group and 7.8 for the STG group. CONCLUSION: In this study, the 2 groups had comparable results in terms of patient satisfaction, activity level, and knee function. Our study showed that patellofemoral problems and loss of knee motion are more frequent in patients with PT grafts than in those with STG grafts. LEVEL OF EVIDENCE: Level I, Randomized Controlled Trial. PMID- 15800521 TI - Revision anterior cruciate ligament reconstruction: three- to nine-year follow up. AB - PURPOSE: With the increasing number of primary anterior cruciate ligament (ACL) reconstructions, revisions are more frequent. The literature quotes inferior results for revision cases when compared with primary ACL reconstruction. The purpose of the study was to review our institution's experience with revision ACL reconstruction. TYPE OF STUDY: Retrospective case series. METHODS: Thirty-five revision cases were performed between 1993 and 1999. Twenty-nine were available for follow-up. Subjective scores were calculated for Lysholm, Tegner, and International Knee Documentation Committee (IKDC) forms. Objective IKDC scores were determined. KT-1000 measurements were performed as well as isokinetic strength testing of quadriceps and hamstrings. Plain film radiographs were obtained to assess degenerative changes. RESULTS: The average patient age at time of revision was 30.2 years, the average time to revision was 56 months, the follow-up from last revision was 67 months. Twenty-two patients had bone-patellar tendon-bone (BPTB) allograft, 6 had contralateral BPTB autograft, and 1 patient had Achilles allograft. Overall, KT-1000 measurement showed an average of 2.78 mm side-to-side difference of displacement. The allograft versus the autograft group was 3.21 mm versus 1.33 mm, respectively. Prerevision data were unavailable. However, all patients had a positive pivot-shift test before revision. Average postrevision Lysholm, Tegner, and subjective IKDC scores were 86.6, 11.86, and 85.86, respectively. Concerning the IKDC objective scores, 15 patients had an A score, 8 had a B score, and 4 had a C score. All 29 patients available for follow up reported that they would have the surgery again. The average strength of quadriceps and hamstrings ranged from 82% to 88% of uninvolved side. CONCLUSIONS: This study provides long-term follow-up with good results for revision ACL reconstruction. Attention to principles when performing revision ACL surgery is critical to provide satisfactory results. LEVEL OF EVIDENCE: Level IV. PMID- 15800522 TI - Intra-articular findings in the reconstructed multiligament-injured knee. AB - PURPOSE: Intra-articular chondral and meniscal injury in the multiligament injured knee has not been examined. The purpose of this study was to determine the pattern of intra-articular chondral and meniscal damage in subjects with multiligament knee injuries undergoing surgery. TYPE OF STUDY: Prospective cohort study. METHODS: Analysis was performed on prospectively collected data for the presence of meniscal tears and significant chondral injury on subjects who underwent knee ligament surgery. Subjects were grouped by their multiligament injury combination. Chondral injury was graded using the Modified Outerbridge Classification system. Criteria for significant chondral defect were any grade 2 lesion involving 50% or more of condylar width and all grade 3 or 4 lesions. Groups with more than 10 subjects were compared against the anterior cruciate ligament (ACL)-only group. RESULTS: Data were collected on 2,265 subjects. The ACL/medial collateral ligament (MCL) injury pattern was the most common multiligament injury, comprising 70.5% of all multiligament injuries. Lateral meniscal damage was significantly higher in the ACL/MCL group (P < .01). Medial meniscal damage was significantly lower in the ACL/MCL group (P < .01). Medial and lateral meniscal damage was significantly lower in the ACL/lateral collateral ligament (LCL) group (P < .01). Articular damage to the medial tibial plateau was significantly lower in the ACL/MCL group (P < .01). All other multiligament injury patterns showed chondral damage similar to the ACL-only group. Ligament injuries repaired acutely had significantly less articular and medial meniscal damage than chronic repairs (P < .01). CONCLUSIONS: The ACL/MCL injury pattern was the most common multiligament injury pattern and showed a high incidence of lateral meniscal tears. ACL/LCL injuries had fewer meniscal injuries. Multiligament-injured knees showed increased risk of a torn medial meniscus as chronicity increased. LEVEL OF EVIDENCE: Level IV. PMID- 15800523 TI - Factors related to additional knee injuries after anterior cruciate ligament injury. AB - PURPOSE: We reviewed 1,375 consecutive patients to determine whether patient gender, age at injury, or activity level at injury were related to the risk of additional knee injuries over time following anterior cruciate ligament (ACL) injury. TYPE OF STUDY: Retrospective case series. METHODS: Survival analysis was used to analyze the effect of each factor while statistically controlling for the effect of time from ACL injury to reconstruction. RESULTS: The risk of meniscus injury was significantly higher among male subjects (odds ratio [OR] = 1.5, P < .001). When undergoing ACL reconstruction more than 6 months after injury, the risk of meniscus injury increased by 1.5 times for male subjects (P = .021) but increased by 3.4 times for female subjects (P < .001). The risk of meniscus injuries was significantly higher among patients undergoing ACL reconstruction more than 6 months after injury when compared with patients undergoing reconstruction within 2 weeks of injury (OR = 2.2, P < .001). The risk of articular cartilage lesions was significantly higher among patients undergoing ACL reconstruction more than 1 year after injury when compared with patients undergoing reconstruction within 2 weeks of injury (OR = 2.1, P < .001). CONCLUSIONS: Men had a consistently higher occurrence rate of meniscus injuries than did women. The risk of meniscus injuries increased at a higher rate over time among women. The risk of meniscus injuries increases when ACL reconstruction is performed more than 6 months after injury. The risk of articular cartilage lesions increases when ACL reconstruction is performed more than 1 year after injury. LEVEL OF EVIDENCE: Level IV, Case Series. PMID- 15800524 TI - Contact pressure in anterior cruciate ligament bone tunnels: comparison of endoscopic and two-incision technique. AB - PURPOSE: To investigate differences in contact pressure in the bone tunnel between the inside-out and outside-in techniques used for anterior cruciate ligament reconstruction. TYPE OF STUDY: In vitro cadaveric analysis. METHODS: Four fresh-frozen cadaver knees were tested. A femoral tunnel was generated through the tibial tunnel (inside-out technique). Another femoral tunnel was created using a rear-entry guide at the same intra-articular exit (outside-in technique). Aluminum cylinders containing 4-way (anterior, posterior, medial, and lateral) conductive rubber pressure sensors at the entrance of the joint were inserted into the tibial and femoral tunnels. Dynamic changes in the contact pressure of grafts in the femoral and tibial tunnels during 0 degrees to 130 degrees of knee flexion were measured. RESULTS: Contact pressure of the graft migrated from the anterior to the posterior portion of the femoral tunnel upon alignment of the femoral and tibial tunnels. At 0 degrees, 30 degrees, and 60 degrees of knee flexion, significantly greater contact pressure was observed at the anterior portion of the femoral tunnel made by the inside-out technique compared with that in the tunnel made by the outside-in technique. At 0 degrees, 30 degrees, and 60 degrees of knee flexion, significantly greater contact pressure was observed at the lateral portion of the femoral tunnel made by the outside-in technique. Constant contact pressure was observed in the medial portion of the tunnel made by the inside-out technique. Pressure changes in the tibial tunnel were slight; moreover, changes were not influenced by the femoral tunnel direction under any conditions. CONCLUSIONS: This study shows that contact pressure in the femoral tunnel is influenced by the direction of the femoral tunnel. CLINICAL RELEVANCE: The femoral tunnel direction in anterior cruciate ligament reconstruction is an important factor for femoral tunnel enlargement. A horizontal direction of the femoral tunnel might be needed for mature bone graft incorporation. PMID- 15800525 TI - Ganglion cysts of the anterior cruciate ligament: a series of 15 cases. AB - PURPOSE: To increase awareness of the presence of ganglion cysts of the anterior cruciate ligament and to consider this diagnosis in any knee that has lost range of motion (ROM) in the absence of osteoarthritis. TYPE OF STUDY: Case series. METHODS: We present a series of 15 cases recorded over a period of 5 years illustrating the clinical presentation and additional pathology seen at arthroscopy. International Knee Documentation Committee scores were calculated at the recent follow-up to establish outcome of the surgery. RESULTS: The cysts occurred predominantly in men with the most common presenting complaint being decreased ROM and pain. Overall, results were excellent with no recurrence of symptoms at a mean 36-month follow-up, but outcome does appear to be associated with other pathology present. ROM was improved with arthroscopic excision of the cyst in all cases that were impaired. CONCLUSIONS: Although rare, ganglion cysts related to the cruciate ligaments of the knee should be considered in the differential diagnosis of a painful knee especially when associated with a decreased ROM and no evidence of osteoarthritis on radiographs. Other pathology can often be present, which may affect the overall outcome, but arthroscopic debridement of the ganglion cyst should be considered the treatment of choice in order to reliably restore active ROM. LEVEL OF EVIDENCE: Level IV, Case Series. PMID- 15800526 TI - The effect of electrothermal shrinkage on the biomechanical properties of the anterior cruciate ligament: an experimental study. AB - PURPOSE: The purpose of this study was to determine the acute effects of electrothermal shrinkage on the biomechanical properties of the anterior cruciate ligament (ACL). TYPE OF STUDY: Randomized trial. METHODS: Fifty fresh femur-ACL tibia complexes harvested from fully matured pigs were used. The femur anteromedial bundle-tibia complex specimens were randomly divided into 5 groups of 10 specimens each. In each group, radiofrequency energy set at nonablative levels was applied to the anteromedial bundle of the ACL with a bipolar radiofrequency generator. In groups I and II, radiofrequency treatment was applied to the ACL using 28 W and 45 W power, respectively, for 30 seconds. In groups III and IV, radiofrequency treatment was applied to the ACL using 28 W and 45 W power, respectively, for 60 seconds. In group V, no treatment was applied in order to obtain normal control data. The treatment was performed in physiological saline solution under a 1-N load. In each group, 8 of the 10 specimens were used for biomechanical evaluation, and the remaining 2 were used for histologic observation. RESULTS: Concerning the length of the anteromedial bundle after the treatment, the analysis of variance (ANOVA) showed a significant difference among the 5 groups. The length in groups I, II, III, and IV was significantly shorter than that of group V. In tensile testing, the stiffness was 120, 116, 113, 89, and 156 N/mm in groups I, II, III, IV, and V, respectively. The ANOVA showed a significant difference among the groups. Groups I, II, III, and IV were significantly lower than group V, respectively. Histologic examination showed diffuse collagenous denaturation and pyknotic nuclear changes in fibroblasts at the treated portion. The collagen crimp pattern was not present in the treated area. CONCLUSIONS: Application of RF energy to the specimens caused both shortening and weakening according to the magnitude and duration of the application. CLINICAL RELEVANCE: These results may explain one of the causes of the poor results reported in some clinical studies. This study warns against a too optimistic application of electrothermal shrinkage to the ACL as a clinical treatment. PMID- 15800527 TI - Subjective results of nonoperatively treated, acute, isolated posterior cruciate ligament injuries. AB - PURPOSE: To evaluate subjective results obtained prospectively of patients who had acute isolated posterior cruciate ligament injuries. TYPE OF STUDY: Prospective cohort study. METHODS: Between 1983 and 2001, 271 patients who were seen for an acute, isolated posterior cruciate ligament (PCL) injury were asked to enroll in a long-term study (grade 1, n = 100; grade 1.5, n = 43; grade 2, n = 128). Patients were evaluated using a modified Noyes subjective knee survey (mailed yearly), an activity survey (since 1998), and the International Knee Documentation Committee (IKDC) Subjective Knee Survey (since 2000). RESULTS: The most recent modified Noyes survey was obtained from 215 patients at a mean time of 7.8 years after injury (range, 1 to 18 years), and the mean total score was 85.6 +/-15.0 points. IKDC subjective scores were obtained from 85 patients at a mean time of 8.8 years after injury, and the mean score was 82.7 +/- 16.0 points. There was a statistically significant correlation between the IKDC subjective score and the modified Noyes total score (R2 = .56426, P < .0001). Patients with greater PCL laxity did not have statistically significant lower subjective scores than patients with lesser PCL laxity. There were 146 patients who had at least 4 modified Noyes subjective surveys that could be evaluated for their consistency of total scores through time after injury. Total scores were consistently excellent for 40%, consistently good for 10%, consistently fair for 6%, consistently poor for 2%, consistently improving scores for 16%, decreasing scores for 12%, and inconsistent scores for 14%. Of 67 patients who scored less than 85 points in the first 2 years after injury, only 34 had a score of less than 85 points at their most recent survey. CONCLUSIONS: Subjective scores of patients with acute, isolated PCL injuries were independent of grade of PCL laxity and mean scores did not decrease with time from injury. No identifiable characteristics were identified that would help determine which patients with isolated PCL injuries would have deteriorating knee function. LEVEL OF EVIDENCE: Level I, Prospective Prognostic Study. PMID- 15800528 TI - Multiple osteochondral arthroscopic grafting (mosaicplasty) for cartilage defects of the knee: prospective study results at 2-year follow-up. AB - PURPOSE: To prospectively evaluate the mosaicplasty technique for treatment of femoral condyle cartilage lesions (Outerbridge grade IV) less than 2.5 cm2 in homogeneous group of young active patients. TYPE OF STUDY: Case series. METHODS: Thirty-seven patients (10 female, 27 male; mean age, 29.5 years) with full thickness knee chondral lesions were treated by the arthroscopic mosaicplasty technique. All patients practiced sports. There were 12 ACL reconstructions, 11 medial meniscectomies, and 8 lateral meniscectomies associated; 10 meniscectomies, 9 ACL reconstructions, and 5 cartilage reparative operations had been previously performed. All patients were evaluated at a 2-year follow-up. The International Cartilage Repair Society (ICRS) form, return to sports, computed tomography, or magnetic resonance imaging were used for clinical evaluation. In some cases, second-look arthroscopy was performed. RESULTS: The ICRS showed 78.3% good and excellent results; 27 patients returned to sports at the same level and 5 at a lower level, but 5 were not able to resume sports. Results in the lateral condyles were significantly better than those in medial condyles, and younger patients had a better clinical outcome than did older patients. Cases with associated surgery had better clinical results; previous surgery did not significantly influence the clinical outcome. CONCLUSIONS: The results of this technique at medium-term follow-up are encouraging with 78.3% clinically satisfactory results. Better results can be obtained in young patients with associated surgery, with localized grade 4 lesions of the lateral condyles. This arthroscopic 1-step surgery appears to be a valid solution for the treatment of grade III-IV cartilage defects not more than 2.5 cm2. LEVEL OF EVIDENCE: Level IV, Case Series. PMID- 15800529 TI - The multiple-ligament injured knee: evaluation, treatment, and results. AB - The multiple-ligament injured knee is a complex problem in orthopaedic surgery. Most dislocated knees involve tears of the anterior and posterior cruciate ligaments (ACL/PCL) and at least 1 collateral ligament complex. Careful assessment of the extremity vascular status is essential because of the possibility of arterial and/or venous compromise. These complex injuries require a systematic approach to evaluation and treatment. Physical examination and imaging studies enable the surgeon to make a correct diagnosis and to formulate a treatment plan. Arthroscopically assisted combined ACL/PCL reconstruction is a reproducible procedure. Knee stability is improved postoperatively when evaluated using knee ligament rating scales, arthrometer testing, and stress radiographic analysis. Acute medial collateral ligament (MCL) tears, when combined with ACL/PCL tears, may in certain cases be treated with bracing. Posterolateral corner injuries combined with ACL/PCL tears are best treated with primary repair as indicated, combined with reconstruction using a post of strong autograft (split biceps tendon, biceps tendon, semitendinosus) or allograft (Achilles tendon, bone-patellar tendon-bone) tissue. Surgical timing depends on the ligaments injured, the vascular status of the extremity, reduction stability, and the overall health of the patient. We prefer to use allograft tissue for reconstruction in these cases because of the strength of these large grafts and the absence of donor site morbidity. PMID- 15800530 TI - Arthroscopic reconstruction of an isolated avulsion fracture of the lesser tuberosity. AB - Isolated avulsion fracture of the lesser tuberosity represents an extremely rare injury that occurs mainly in younger patients. This report outlines the case of a 35-year-old healthy man who sustained a bony avulsion of the subscapularis tendon during an abduction external rotation trauma to his left shoulder. The injury was diagnosed by physical examination, standard radiographs, and magnetic resonance imaging. An arthroscopic reconstruction including closed reduction and internal fixation using suture anchors was performed. After a follow-up of 6 months, the patient's Constant Score increased from 62.4 points preoperatively to 91.3 points. At the latest follow-up, the patient was completely pain free, had regained a full work activity level, and returned to overhead sports. On postoperative standard radiographs and magnetic resonance images, the bony fragment was shown to be consolidated in an anatomic position. In cases of displaced isolated fractures of the lesser tuberosity without significant subluxation or dislocation of the long head of the biceps arthroscopic reconstruction using suture anchors can lead to a good clinical and radiological result. PMID- 15800531 TI - Disintegration of an absorbable rotator cuff anchor six weeks after implantation. AB - Rotator cuff failure by suture-bone or suture anchor pull-out, suture breakage, knot slippage, and tendon pull-out are well described. I report a case of early disintegration of a bioabsorbable suture anchor. A 77-year-old woman underwent arthroscopic rotator cuff repair. On suspecting failure, the repair was repeated 40 days later. Arthroscopy revealed disintegration of the suture loop from the anchor. Open rotator cuff repair was then performed with transosseous suture and metallic anchors. PMID- 15800532 TI - The HAGL lesion: an arthroscopic technique for repair of humeral avulsion of the glenohumeral ligaments. AB - Glenoid avulsion of the capsulolabral complex and associated capsular laxity are well-described results of traumatic anterior dislocation. A less common consequence of traumatic anterior instability is the humeral avulsion of the glenohumeral ligaments (HAGL) lesion. An understanding of the arthroscopic pathoanatomy of the HAGL lesion will assist the surgeon in recognizing this uncommon entity. We describe a suture anchor technique for arthroscopic repair of HAGL lesions that anatomically reapproximates the torn edge of the glenohumeral ligament complex to its humeral head insertion. The technique is technically straightforward and does not require special equipment beyond that typical for arthroscopic labral repairs. PMID- 15800533 TI - Lateral elbow reconstruction using a new fixation technique. AB - A radial collateral ligament reconstruction using interference fit screws instead of bone tunnels is described that uses a new screw design. A standard free graft is harvested. The isometric points on the lateral epicondylar ridge and ulna are identified. Fixation is completed after the isometricity of the insertion site is ensured and the graft is appropriately tensioned. A new interference-fit screw system is used (Arthrex, Naples, FL). There is much less trauma to the elbow, the fixation is stronger, and the scarring I believe will be minimized based on this technique. PMID- 15800534 TI - A new arthroscopic technique for revision of the posterior compartment in symptomatic total knee arthroplasty. AB - Arthroscopy is effective in treating the symptomatic knee following total knee arthroplasty. A small number of patients complain about postoperative pain in the posterior compartment. Loose bodies and degenerated remnants of the menisci are the main reasons for this pain. We present a new technique for diagnostic and therapeutic arthroscopy of the posterior compartments in patients with symptomatic total knee arthroplasty. After standard anterior arthroscopy, a wide circular notchplasty is performed to allow easy access to the posterior lateral compartment of the knee joint and to apply a posterolateral working portal under direct arthroscopic control. If inspection of the posteromedial compartment is necessary, the loose tissue behind the posterior cruciate ligament is resected to gain access to the posteromedial compartment. PMID- 15800549 TI - What to tell your clients about eating fish. PMID- 15800550 TI - The request for proposal: a primer. PMID- 15800551 TI - Improving nutrition for American seniors: a new look at the Older Americans Act. PMID- 15800552 TI - Genetic dietetics: nutrigenomics and the future of dietetics practice. PMID- 15800553 TI - Dietary intake in human immunodeficiency virus-infected adults: a comparison of dietary assessment methods. AB - OBJECTIVE: The objective of this study was to compare estimated nutrient intakes from 3-day food records vs Food Frequency Questionnaire (FFQ) in a large cohort of individuals living with human immunodeficiency virus (HIV) infection. DESIGN: Dietary data from 315 HIV-positive participants enrolled in a longitudinal cohort study were collected. Nutrient intake data estimated from the Block FFQ were compared with multiple 3-day food records done over the same time period. SUBJECTS/SETTING: Participants enrolled in Nutrition for Healthy Living, an ongoing cohort study based in Boston, MA, were studied. STATISTICAL ANALYSES PERFORMED: Using 3-day food records as the gold standard, we compared estimated nutrient intakes between the two assessment methods by (a) median of individual differences in intake, (b) correlation coefficients, (c) quintile ranking, and (d) intakes less than the Dietary Reference Intakes. Nutrient intake estimates between the two methods were compared using Spearman, Pearson, and deattenuated correlation coefficients. RESULTS: Median reported intakes of all macronutrients and most micronutrients were significantly less on the FFQ for both men and women. Deattenuated correlations were less than 0.5 for 86% of the men and for 68% of the women. Mean percent agreement for quintile ranking was 30%. CONCLUSIONS: In this large cohort of HIV-infected adults, we found significant differences between FFQs and food records in estimates of absolute dietary intakes, correlation coefficients, quintile rankings, and proportions of subjects consuming less than the Dietary Reference Intakes. These findings have implications for the interpretation and application of dietary intake data from FFQs in specialized populations. PMID- 15800554 TI - Parental pressure, dietary patterns, and weight status among girls who are "picky eaters". AB - OBJECTIVE: To determine whether mothers' fruit and vegetable intake and mothers' use of pressure in the feeding domain when their daughters were 7 years old predicted picky eating and dietary intake when their daughters were 9 years old, and to examine diet and weight status in picky and nonpicky eaters. DESIGN/SUBJECTS: Participants were 173 9-year-old non-Hispanic white girls and their mothers. A longitudinal analysis was used to assess maternal influences on picky eating and diet. A cross-sectional analysis was used to examine diet and weight status between picky and nonpicky eaters. Measures included maternal feeding practices, daughters' pickiness, mothers' fruit and vegetable intake, daughters' food intake, and weight status. STATISTICAL ANALYSES: t tests examined differences between picky and nonpicky eaters. Structural equation modeling examined relationships among mothers' fruit and vegetable intake; child feeding practices; daughters' pickiness; and fruit, vegetable, micronutrient, and fiber intakes. RESULTS: Mothers consuming more fruits and vegetables were less likely to pressure their daughters to eat and had daughters who were less picky and consumed more fruits and vegetables. Picky eaters consumed fewer fruits and vegetables, but also fewer fats and sweets. All girls consumed low amounts of vitamin E, calcium, and magnesium, but more picky girls were at risk for not meeting recommendations for vitamins E and C and also consumed significantly less fiber. In addition, picky eaters were less likely to be overweight. CONCLUSIONS: Mothers influenced daughters' fruit and vegetable intake via their own patterns of fruit and vegetable intake and by influencing their daughters' tendencies to be picky eaters. Both picky and nonpicky eaters had aspects of their diets that did not meet recommendations. Taken together, these findings suggest that parents should focus less on "picky eating" behavior and more on modeling fruit and vegetable consumption for their children. PMID- 15800555 TI - Assessment of nutritional status in adult patients with cystic fibrosis: whole body bioimpedance vs body mass index, skinfolds, and leg-to-leg bioimpedance. AB - OBJECTIVE: To investigate whether body mass index (BMI) or body fat percentage estimated from BMI, skinfolds, or leg-to-leg bioimpedance are good indicators of nutritional status in adult patients with cystic fibrosis. Body fat percentage measured by whole-body bioimpedance was used as the reference method. DESIGN: Cross-sectional study using four methods to estimate body fat percentage. All patients filled out a food frequency and a physical activity questionnaire for assessment of their habitual food intake and energy requirements, respectively. SUBJECTS/SETTING: Thirty-five adult patients (23 men/12 women) with cystic fibrosis, age range 18 to 46 years, were measured during their yearly visit at the outpatient clinic of the Cystic Fibrosis Center in Utrecht, the Netherlands. STATISTICAL ANALYSIS: Mean+/-standard deviation was calculated for all measurements and 95% confidence intervals for differences between methods. Bland Altman plots were used to assess differences between the measures of body composition and Pearson correlation coefficients were calculated to determine the relationships between them, and between the energy requirements and the energy intakes. RESULTS: For men the whole-body body fat percentage reference was 14.1%+/-3.0, body fat percentage estimated from BMI was 15.8%+/-4.3, body fat percentage estimated from skinfolds was 8.6%+/-4.8, and body fat percentage estimated from leg-to-leg bioimpedance was 13.1%+/-4.9. For women the whole-body body fat percentage reference was 24.0%+/-5.9, body fat percentage estimated from BMI was 25.1%+/-4.0, body fat percentage estimated from skinfolds was 17.0%+/ 4.8, and body fat percentage estimated from leg-to-leg bioimpedance was 25.0%+/ 6.9. Body fat percentage estimated from BMI and body fat percentage estimated from skinfolds were significantly different from the reference value for body fat percentage (P <.05). The correlation coefficients between the reference body fat percentage and body fat percentage estimated from BMI, from skinfolds, and from leg-to-leg bioimpedance were all more than 0.72. In all but one patient, nutritional status was correctly assessed by BMI: those with a BMI less than 18.5 had body fat percentage less than 10% (men) or less than 20% (women). The mean energy intake of the men was 141% of the Recommended Dietary Allowance as proposed in European and Dutch guidelines. The mean energy intake of the women was 94% of the Recommended Dietary Allowance. CONCLUSIONS: A simple calculation of BMI is adequate to diagnose nutritional status in adult patients with cystic fibrosis. Bioimpedance measurements are only needed when nutritional therapy specifically focuses on lean body mass. PMID- 15800557 TI - Glycemic index of potatoes commonly consumed in North America. AB - OBJECTIVE: To determine the effect of variety and cooking method on glycemic response and glycemic index of common North American potatoes. DESIGN: Study 1: subjects consumed 200 g Russet or white potatoes that were either (a) precooked, refrigerated, and reheated (precooked) or (b) cooked and consumed immediately (day-cooked). Incremental area under the curve was determined. Study 2: subjects consumed 50 g carbohydrate portions of white bread or potatoes (six different varieties and two different cooking methods). Glycemic index values were calculated. In both studies meals were consumed after a 10- to 12-hour overnight fast and finger-prick capillary-blood glucose was measured before and at intervals for 2 hours after consumption. SUBJECTS: The study groups were as follows: Study 1 comprised four men and six women, aged 20 to 44 and Study 2 comprised 11 men and one woman, aged 18 to 50. STATISTICAL ANALYSES: Repeated measures analysis of variance with Newman-Kuels to protect for multiple comparisons (criterion of significance two-tailed P <.05). RESULTS: Study 1: Precooked Russet potatoes elicited lower area under the curve than day-cooked (P <.05), while precooking had no effect on boiled white potatoes. Study 2: The glycemic index values of potatoes varied significantly, depending on the variety and cooking method used (P =.003) ranging from intermediate (boiled red potatoes consumed cold: 56) to moderately high (roasted California white potatoes: 72; baked US Russet potatoes: 77) to high (instant mashed potatoes: 88; boiled red potatoes: 89). CONCLUSIONS: The glycemic index of potatoes is influenced by variety and method of cooking and US Russet potatoes have only a moderately high glycemic index. Individuals who wish to minimize dietary glycemic index can be advised to precook potatoes and consume them cold or reheated. PMID- 15800558 TI - Patients receiving maintenance hemodialysis with low vs high levels of nutritional risk have decreased morbidity. AB - OBJECTIVE: To compare the demographic and clinical characteristics and outcomes (morbidity) of 442 patients receiving maintenance hemodialysis who are at different levels of nutritional risk. DESIGN: A retrospective, longitudinal, chart review. SETTING/SUBJECTS: An urban, outpatient hemodialysis unit in New York City. Subjects were stratified according to their number of nutritional risk factors: zero to one=low risk, two to three=moderate risk, four to six=high risk. MAIN OUTCOME MEASURES: Mean values for serum albumin <37 g/L, creatinine <884 micromol/L, total cholesterol <4.42 mmol/L, normalized protein nitrogen appearance <0.9 g/kg/day, weight change > -2.5 kg, and body mass index <24. Morbidity indicators were frequency and duration of hospitalizations. STATISTICAL ANALYSES: Descriptive statistics, analysis of variance, and chi 2 analysis were used to summarize data and to analyze mean differences between the groups and differences in categorical variables, respectively. RESULTS: Compared with the high-risk group, the majority of subjects in the low-risk group were younger, male, and did not have diabetes; fewer had two or more comorbidities. The high risk group had 75% more hospitalizations and spent 195% more days in the hospital than the low-risk group. CONCLUSIONS: Declining values of the nutritional risk factors and higher hospitalization rates were present in the high-risk group. Older subjects, those with diabetes, and those with two or more comorbidities comprised the majority of the high-risk group. More aggressive nutrition counseling and interventions may be needed for high-risk group members to determine if their risk for morbidity could be reduced. PMID- 15800559 TI - Effect of high protein vs high carbohydrate intake on insulin sensitivity, body weight, hemoglobin A1c, and blood pressure in patients with type 2 diabetes mellitus. AB - BACKGROUND: Extremely low carbohydrate/high protein diets are popular methods of weight loss. Compliance with these diets is poor and long-term effectiveness and the safety of these diets for patients with type 2 diabetes is not known. OBJECTIVE: The objective of the current study was to evaluate effects of less extreme changes in carbohydrate or protein diets on weight, insulin sensitivity, glycemic control, cardiovascular risk factors (blood pressure, lipid levels), and renal function in obese inner-city patients with type 2 diabetes. DESIGN: Study patients were admitted to the General Clinical Research Center for 24 hours for initial tests including a hyperinsulinemic-euglycemic clamp (for measurement of insulin sensitivity), bioelectrical impedance analysis (BIA) and anthropometric measurements (for assessment of body composition), indirect calorimetry (for measurement of REE), electronic blood pressure monitoring, and blood chemistries to measure blood lipids levels along with renal and hepatic functions. Six patients with type 2 diabetes (five women and one man) were randomly assigned to the high-protein diet (40% carbohydrate, 30% protein, 30% fat) and six patients (four women and two men) to the high-carbohydrate diet (55% carbohydrate, 15% protein, 30% fat). All patients returned to the General Clinical Research Center weekly for monitoring of food records; dietary compliance; and measurements of body weight, blood pressure, and blood glucose. After 8 weeks on these diets, all patients were readmitted to the General Clinical Research Center for the same series of tests. INTERVENTION: Twelve study patients were taught to select either the high-protein or high-carbohydrate diet and were followed for 8 weeks. MAIN OUTCOME MEASURES: Insulin sensitivity, hemoglobin A1c, weight, and blood pressure were measured. STATISTICAL ANALYSES: Statistical significance was assessed using two-tailed Student's t tests and two-way repeated measures analysis of variance. RESULTS: Both the high-carbohydrate and high-protein groups lost weight (-2.2+/ 0.9 kg, -2.5+/-1.6 kg, respectively, P <.05) and the difference between the groups was not significant (P =.9). In the high-carbohydrate group, hemoglobin A1c decreased (from 8.2% to 6.9%, P <.03), fasting plasma glucose decreased (from 8.8 to 7.2 mmol/L, P <.02), and insulin sensitivity increased (from 12.8 to 17.2 micromol/kg/min, P <.03). No significant changes in these parameters occurred in the high-protein group, instead systolic and diastolic blood pressures decreased (-10.5+/-2.3 mm Hg, P =.003 and -18+/-9.0 mm Hg, P <.05, respectively). After 2 months on these hypocaloric diets, each diet had either no or minimal effects on lipid levels (total cholesterol, low-density lipoprotein, high-density lipoprotein), renal (blood urea nitrogen, serum creatinine), or hepatic function (aspartate aminotransferase, alanine aminotransferase, bilirubin). PMID- 15800560 TI - Nutrient distribution for type 2 diabetes: what's a dietetics professional to do? PMID- 15800561 TI - Incorporating human genetics into dietetics curricula remains a challenge. AB - This descriptive survey was undertaken to assess integration of genetics into undergraduate didactic dietetic education. The response rate was 35% (n=82) of all directors (N=232) of accredited or approved Didactic Programs in Dietetics. Although most directors (n=58 of 82) agreed that genetics is an important component of dietetics education, they expressed concerns about already-crowded curricula and lack of time, resources, and knowledge. Thirty-eight directors indicated that they had no familiarity with the core competencies in genetics for all health professionals. Genetics is included in some way in 69 of the 82 programs that responded. Courses in which genetics was most likely to be incorporated included nutrition, physiology, microbiology, and biochemistry. Only four directors reported a required course entirely devoted to genetics. Programs were most likely to meet the knowledge competency of the role of genetic factors in maintaining health and preventing disease and least likely to address the genetic counseling process and indications for referral to specialists. Applications of genetics in dietetics will continue to grow in importance. Evidence from this study indicates that current curricula provide little to no genetics content. Nutrition faculty must become more knowledgeable about genetics before being expected to increase genetics content in entry-level dietetics curricula. PMID- 15800562 TI - Nutritional genomics in practice: where do we begin? AB - Nutritional genomics, which studies the genome-wide influences of nutrition, has far-reaching potential in the prevention of diet-related disease. It is highly likely that during the next decade the nutritional supplement and functional food industries will continue robust growth in response to advances in nutritional genomics research and its applications. Parallel to this growth will be impressive progress in understanding the specific influence of certain food components on metabolic pathways and on long-term risk for disease. As genetic information about individuals becomes available, such data are likely to redefine the current concept of preventive medicine. Dietetics professionals have the potential to harness this information and influence health promotion and disease prevention on a global scale. For these reasons, the dietetics profession has an exciting opportunity that, if seized and properly executed, could enhance the scientific foundation of clinical practice, improve therapeutic outcomes, and significantly expand career and economic opportunities for practitioners. The future of dietetics is unquestionably intertwined with nutritional genomics. PMID- 15800563 TI - College athletes with an elevated body mass index often have a high upper arm muscle area, but not elevated triceps and subscapular skinfolds. AB - Body mass index (BMI, calculated as kg/m 2 ) is increased by high amounts of both lean and fat tissue. Therefore, a very muscular individual with low body fat could be classified as overweight by BMI. To evaluate this problem, the relationship between BMI, body fat as indicated by the sum of triceps and subscapular skinfolds, and muscle mass as indicated by upper arm muscle area (UAMA) was studied in 107 male and 106 female National Collegiate Athletic Association Division III athletes. Sex, the sum of triceps and subscapular skinfolds, and UAMA were significantly related to BMI (overall P <.0001, R 2 =0.617), although the relationship was nonlinear and more complex in females than in males. Thirty-eight athletes had a BMI of 25 or higher, indicating overweight or obesity. Of these, only four had excess body fat, as indicated by a sum of triceps and subscapular skinfolds greater than the 85th percentile, but 27 had high muscle mass, indicated by a UAMA greater than the 85th percentile. In the nonrandom sample of athletes we studied, BMI frequently classified muscular individuals who did not have high skinfold measurements as overweight. PMID- 15800564 TI - Nutrition concerns in discharge planning for older adults: a need for multidisciplinary collaboration. AB - This survey of 84 case managers (CMs) (88% response rate) in 11 south Florida hospitals, in one for-profit health care system, sought to identify their discharge planning concerns regarding the food and nutrition needs of older patients, how they addressed these concerns, and the degree to which registered dietitians (RDs) were involved. Most CMs were female (82 of 84, 98%), older than age 40 (59 of 84, 70%), and were registered nurses (51 of 84, 61%). Almost all (82 of 84, 98%) reported job barriers, including excessive patient loads and responsibilities and limited community services. Almost all said that nutrition related diseases and factors (eg, chewing/swallowing problems, poor appetite, modified diets, poor dentition) strongly influenced discharge planning. Many perceived community nutrition resources (eg, congregate meals, food stamps, shopping assistance, outpatient dietitians) as not readily available. While physicians, nurses, social workers, and physical therapists were identified as very important in discharge planning, RDs were not; almost half of CMs consulted them infrequently, if at all. Strategies for the six nutrition-related case scenarios were inconsistent. Home-health agencies were chosen most often and outpatient RDs least often. Comprehensive discharge planning must include more attention to nutrition with greater input from clinical, outpatient, home health, and community RDs through, for example, attendance at hospital discharge planning rounds, inservices for CMs, and better marketing of RD services. More RDs can and should become CMs to help serve the number of older adults with nutrition-related chronic conditions. PMID- 15800565 TI - Urinary ketones reflect serum ketone concentration but do not relate to weight loss in overweight premenopausal women following a low-carbohydrate/high-protein diet. AB - This study examined the effect of a low-carbohydrate/high-protein diet on serum and urine ketone body concentrations. Thirteen overweight premenopausal women aged 32 to 45 years consumed < or =20 g carbohydrate/day with liberal intakes of protein and fat for 2 weeks; thereafter, carbohydrate intake increased 5 g/week for 10 weeks. Women were weighed and provided fasting urine and blood samples to detect urinary ketones and quantify serum ketone concentrations, respectively, at baseline and weeks 1 to 4, 6, and 12. Women lost 8.3%+/-2.8% of initial body weight by week 12. Serum beta-hydroxybutyrate production was highest at week 1 and declined weekly, with all values higher than baseline (P <.05). Each week, serum beta-hydroxybutyrate was correlated with presence of urinary ketones (P <.05), but no relationship was found between weekly weight change and serum ketone production. Urinary ketones are detected in premenopausal women complying with a low-carbohydrate/high-protein diet and are associated with serum ketone concentration. However, serum ketones do not reflect weight loss. PMID- 15800566 TI - Resting energy expenditure is lower than predicted in people taking atypical antipsychotic medication. AB - Resting energy expenditure (REE) is lower than predicted in persons taking atypical antipsychotic medication, and weight management is a significant clinical challenge for some of them. However, to date there have been no published guidelines to assist clinicians in choosing appropriate prediction equations to estimate energy expenditure in persons taking atypical antipsychotic medications. The objectives of this study were to measure REE in a group of men taking the atypical antipsychotic clozapine and to determine whether REE can be accurately predicted for this population using previously published regression equations. REE was measured using indirect calorimetry via a ventilated hood on eight men who had completed at least 6 months of treatment with clozapine. Comparisons between measured REE and predicted REE using five different equations were undertaken. The commonly-used Harris-Benedict and Schofield equations systematically overestimated REE. Predictions of REE from other equations were too variable for clinical use. When estimating energy requirements as part of a weight-management program in men who have been taking clozapine for 6 months, predictions of REE from the equations of Harris-Benedict and Schofield should be reduced by 280 kcal/day. PMID- 15800567 TI - Position paper of the American Dietetic Association: nutrition across the spectrum of aging. AB - It is the position of the American Dietetic Association that older Americans receive appropriate care; have broadened access to coordinated, comprehensive food and nutrition services; and receive the benefits of ongoing research to identify the most effective food and nutrition programs, interventions, and therapies across the spectrum of aging. Food and water and nutritional well-being are essential to the health, self-sufficiency, and quality of life for the fast growing, heterogeneous, multiracial, and ethnic populations of older adults. Many people, as they age, remain fully independent and actively engaged in their communities; however, others fare less well and need more support. A broad array of appropriate, culturally sensitive food and nutrition services, physical activities, and health and supportive care customized to the population of older adults are necessary. National, state, and local policies that promote coordination and integration of food and nutrition services into health and supportive systems are needed to maintain independence, functional ability, chronic disease management, and quality of life. Dietetics professionals can take the lead by researching and developing national, state, and local collaborative networks to incorporate effectively the food and nutrition services across the spectrum of aging. PMID- 15800568 TI - American Dietetic Association: scope of dietetics practice framework. PMID- 15800569 TI - American Dietetic Association: standards of practice in nutrition care and updated standards of professional performance. PMID- 15800573 TI - What are the calorie requirements for patients with cystic fibrosis? PMID- 15800578 TI - A state of ignorance. PMID- 15800576 TI - IRF-7 is the master regulator of type-I interferon-dependent immune responses. AB - The type-I interferon (IFN-alpha/beta) response is critical to immunity against viruses and can be triggered in many cell types by cytosolic detection of viral infection, or in differentiated plasmacytoid dendritic cells by the Toll-like receptor 9 (TLR9) subfamily, which generates signals via the adaptor MyD88 to elicit robust IFN induction. Using mice deficient in the Irf7 gene (Irf7-/- mice), we show that the transcription factor IRF-7 is essential for the induction of IFN-alpha/beta genes via the virus-activated, MyD88-independent pathway and the TLR-activated, MyD88-dependent pathway. Viral induction of MyD88-independent IFN-alpha/beta genes is severely impaired in Irf7-/- fibroblasts. Consistently, Irf7-/- mice are more vulnerable than Myd88-/- mice to viral infection, and this correlates with a marked decrease in serum IFN levels, indicating the importance of the IRF-7-dependent induction of systemic IFN responses for innate antiviral immunity. Furthermore, robust induction of IFN production by activation of the TLR9 subfamily in plasmacytoid dendritic cells is entirely dependent on IRF-7, and this MyD88-IRF-7 pathway governs the induction of CD8+ T-cell responses. Thus, all elements of IFN responses, whether the systemic production of IFN in innate immunity or the local action of IFN from plasmacytoid dendritic cells in adaptive immunity, are under the control of IRF-7. PMID- 15800579 TI - Drug safety on trial. PMID- 15800580 TI - Indonesia spared tsunami as disaster quake strikes. PMID- 15800581 TI - Millennium group nails down the financial value of ecosystems. PMID- 15800582 TI - Deal paves way for Congress vote on stem-cell rules. PMID- 15800583 TI - Stray seeds had antibiotic-resistance genes. PMID- 15800584 TI - WWF warns that China's forests are not out of the woods. PMID- 15800585 TI - Journals lack explicit policies for separating eds from ads. PMID- 15800586 TI - Viennese lab renovations stall as cash goes unspent. PMID- 15800587 TI - Biologists snub 'kangaroo court' for Darwin. PMID- 15800588 TI - Political aide sails into top job at maritime institute. PMID- 15800589 TI - Space agencies mull over mutual mission to Europa. PMID- 15800591 TI - Drug safety special: the safety catch. PMID- 15800592 TI - Drug safety special: chasing shadows. PMID- 15800593 TI - Authors say that they prefer online submission. PMID- 15800594 TI - Driving passion brought rare bird to the masses. PMID- 15800595 TI - We Africans must take our future in our hands. PMID- 15800596 TI - Time to speak up for climate-change science. PMID- 15800597 TI - Confronting the human dilemma. PMID- 15800602 TI - The Janus face of Mnemosyne. PMID- 15800603 TI - Structural biology: DNA search and rescue. PMID- 15800604 TI - Techniques: spectroscopy at a stretch. PMID- 15800605 TI - Evolutionary biology: why sex is good. PMID- 15800606 TI - Aceh-Andaman earthquake: what happened and what's next? PMID- 15800607 TI - Cell biology: the more MAD, the merrier. PMID- 15800608 TI - Cosmochemistry: a breath of solar air. PMID- 15800611 TI - Seismology: speed and size of the Sumatra earthquake. AB - Our seismological results reveal that Indonesia's devastating Sumatra-Andaman earthquake on 26 December 2004 was 2.5 times larger than initial reports suggested--second only to the 1960 Chilean earthquake in recorded magnitude. They indicate that it slowly released its energy by slip along a 1,200-km fault, generating a long rupture that contributed to the subsequent tsunami. Now that the entire rupture zone has slipped, the strain accumulated from the subduction of the Indian plate beneath the Burma microplate has been released, and there is no immediate danger of a similar tsunami being generated on this part of the plate boundary, although large earthquakes on segments to the south still present a threat. PMID- 15800609 TI - Biochemistry: a pore way to die. PMID- 15800612 TI - Seismology: energy radiation from the Sumatra earthquake. AB - We determined the duration of high-frequency energy radiation from Indonesia's great Sumatra-Andaman earthquake (26 December 2004) to be about 500 seconds. This duration can be translated into a rupture length of about 1,200 km, which is more than twice as long as that inferred from body-wave analyses performed soon after the event. Our analysis was able rapidly to define the extent of rupture, thereby aiding the assessment of seismic hazard in the immediate future. PMID- 15800613 TI - Chromatin remodelling and epigenetic features of germ cells. AB - Germ cells have the unique capacity to start a new life upon fertilization. They are generated during a sex-specific differentiation programme called gametogenesis. Maturation of germ cells is characterized by an impressive degree of cellular restructuring and gene regulation that involves remarkable genomic reorganization. These events are finely tuned, but are also susceptible to the introduction of various types of error. Because stable genetic transmission to future generations is essential for life, understanding the control of these processes has far-reaching implications for human health and reproduction. PMID- 15800614 TI - An olivine-free mantle source of Hawaiian shield basalts. AB - More than 50 per cent of the Earth's upper mantle consists of olivine and it is generally thought that mantle-derived melts are generated in equilibrium with this mineral. Here, however, we show that the unusually high nickel and silicon contents of most parental Hawaiian magmas are inconsistent with a deep olivine bearing source, because this mineral together with pyroxene buffers both nickel and silicon at lower levels. This can be resolved if the olivine of the mantle peridotite is consumed by reaction with melts derived from recycled oceanic crust, to form a secondary pyroxenitic source. Our modelling shows that more than half of Hawaiian magmas formed during the past 1 Myr came from this source. In addition, we estimate that the proportion of recycled (oceanic) crust varies from 30 per cent near the plume centre to insignificant levels at the plume edge. These results are also consistent with volcano volumes, magma volume flux and seismological observations. PMID- 15800615 TI - CDK-dependent phosphorylation of BRCA2 as a regulatory mechanism for recombinational repair. AB - Inherited mutations in BRCA2 are associated with a predisposition to early-onset breast cancers. The underlying basis of tumorigenesis is thought to be linked to defects in DNA double-strand break repair by homologous recombination. Here we show that the carboxy-terminal region of BRCA2, which interacts directly with the essential recombination protein RAD51, contains a site (serine 3291; S3291) that is phosphorylated by cyclin-dependent kinases. Phosphorylation of S3291 is low in S phase when recombination is active, but increases as cells progress towards mitosis. This modification blocks C-terminal interactions between BRCA2 and RAD51. However, DNA damage overcomes cell cycle regulation by decreasing S3291 phosphorylation and stimulating interactions with RAD51. These results indicate that S3291 phosphorylation might provide a molecular switch to regulate RAD51 recombination activity, providing new insight into why BRCA2 C-terminal deletions lead to radiation sensitivity and cancer predisposition. PMID- 15800616 TI - Structure of a repair enzyme interrogating undamaged DNA elucidates recognition of damaged DNA. AB - How DNA repair proteins distinguish between the rare sites of damage and the vast expanse of normal DNA is poorly understood. Recognizing the mutagenic lesion 8 oxoguanine (oxoG) represents an especially formidable challenge, because this oxidized nucleobase differs by only two atoms from its normal counterpart, guanine (G). Here we report the use of a covalent trapping strategy to capture a human oxoG repair protein, 8-oxoguanine DNA glycosylase I (hOGG1), in the act of interrogating normal DNA. The X-ray structure of the trapped complex features a target G nucleobase extruded from the DNA helix but denied insertion into the lesion recognition pocket of the enzyme. Free energy difference calculations show that both attractive and repulsive interactions have an important role in the preferential binding of oxoG compared with G to the active site. The structure reveals a remarkably effective gate-keeping strategy for lesion discrimination and suggests a mechanism for oxoG insertion into the hOGG1 active site. PMID- 15800617 TI - A non-terrestrial 16O-rich isotopic composition for the protosolar nebula. AB - The discovery in primitive components of meteorites of large oxygen isotopic variations that could not be attributed to mass-dependent fractionation effects has raised a fundamental question: what is the composition of the protosolar gas from which the host grains formed? This composition is probably preserved in the outer layers of the Sun, but the resolution of astronomical spectroscopic measurements is still too poor to be useful for comparison with planetary material. Here we report a precise determination of the oxygen isotopic composition of the solar wind from particles implanted in the outer hundreds of nanometres of metallic grains in the lunar regolith. These layers of the grains are enriched in 16O by >20 +/- 4 per thousand relative to the Earth, Mars and bulk meteorites, which implies the existence in the solar accretion disk of reactions--as yet unknown--that were able to change the 17O/16O and 18O/16O ratios in a way that was not dependent strictly on the mass of the isotope. Photochemical self-shielding of the CO gas irradiated by ultraviolet light may be one of these key processes, because it depends on the abundance of the isotopes, rather than their masses. PMID- 15800618 TI - Unconventional superconductivity in PuCoGa5. AB - In the Bardeen-Cooper-Schrieffer theory of superconductivity, electrons form (Cooper) pairs through an interaction mediated by vibrations in the underlying crystal structure. Like lattice vibrations, antiferromagnetic fluctuations can also produce an attractive interaction creating Cooper pairs, though with spin and angular momentum properties different from those of conventional superconductors. Such interactions have been implicated for two disparate classes of materials--the copper oxides and a set of Ce- and U-based compounds. But because their transition temperatures differ by nearly two orders of magnitude, this raises the question of whether a common pairing mechanism applies. PuCoGa5 has a transition temperature intermediate between those classes and therefore may bridge these extremes. Here we report measurements of the nuclear spin-lattice relaxation rate and Knight shift in PuCoGa5, which demonstrate that it is an unconventional superconductor with properties as expected for antiferromagnetically mediated superconductivity. Scaling of the relaxation rates among all of these materials (a feature not exhibited by their Knight shifts) establishes antiferromagnetic fluctuations as a likely mechanism for their unconventional superconductivity and suggests that related classes of exotic superconductors may yet be discovered. PMID- 15800619 TI - Two-dimensional spectroscopy of electronic couplings in photosynthesis. AB - Time-resolved optical spectroscopy is widely used to study vibrational and electronic dynamics by monitoring transient changes in excited state populations on a femtosecond timescale. Yet the fundamental cause of electronic and vibrational dynamics--the coupling between the different energy levels involved- is usually inferred only indirectly. Two-dimensional femtosecond infrared spectroscopy based on the heterodyne detection of three-pulse photon echoes has recently allowed the direct mapping of vibrational couplings, yielding transient structural information. Here we extend the approach to the visible range and directly measure electronic couplings in a molecular complex, the Fenna-Matthews Olson photosynthetic light-harvesting protein. As in all photosynthetic systems, the conversion of light into chemical energy is driven by electronic couplings that ensure the efficient transport of energy from light-capturing antenna pigments to the reaction centre. We monitor this process as a function of time and frequency and show that excitation energy does not simply cascade stepwise down the energy ladder. We find instead distinct energy transport pathways that depend sensitively on the detailed spatial properties of the delocalized excited state wavefunctions of the whole pigment-protein complex. PMID- 15800620 TI - Decline of the marine ecosystem caused by a reduction in the Atlantic overturning circulation. AB - Reorganizations of the Atlantic meridional overturning circulation were associated with large and abrupt climatic changes in the North Atlantic region during the last glacial period. Projections with climate models suggest that similar reorganizations may also occur in response to anthropogenic global warming. Here I use ensemble simulations with a coupled climate-ecosystem model of intermediate complexity to investigate the possible consequences of such disturbances to the marine ecosystem. In the simulations, a disruption of the Atlantic meridional overturning circulation leads to a collapse of the North Atlantic plankton stocks to less than half of their initial biomass, owing to rapid shoaling of winter mixed layers and their associated separation from the deep ocean nutrient reservoir. Globally integrated export production declines by more than 20 per cent owing to reduced upwelling of nutrient-rich deep water and gradual depletion of upper ocean nutrient concentrations. These model results are consistent with the available high-resolution palaeorecord, and suggest that global ocean productivity is sensitive to changes in the Atlantic meridional overturning circulation. PMID- 15800621 TI - Structural and temporal requirements for geomagnetic field reversal deduced from lava flows. AB - Reversals of the Earth's magnetic field reflect changes in the geodynamo--flow within the outer core--that generates the field. Constraining core processes or mantle properties that induce or modulate reversals requires knowing the timing and morphology of field changes that precede and accompany these reversals. But the short duration of transitional field states and fragmentary nature of even the best palaeomagnetic records make it difficult to provide a timeline for the reversal process. 40Ar/39Ar dating of lavas on Tahiti, long thought to record the primary part of the most recent 'Matuyama-Brunhes' reversal, gives an age of 795 +/- 7 kyr, indistinguishable from that of lavas in Chile and La Palma that record a transition in the Earth's magnetic field, but older than the accepted age for the reversal. Only the 'transitional' lavas on Maui and one from La Palma (dated at 776 +/- 2 kyr), agree with the astronomical age for the reversal. Here we propose that the older lavas record the onset of a geodynamo process, which only on occasion would result in polarity change. This initial instability, associated with the first of two decreases in field intensity, began approximately 18 kyr before the actual polarity switch. These data support the claim that complete reversals require a significant period for magnetic flux to escape from the solid inner core and sufficiently weaken its stabilizing effect. PMID- 15800622 TI - Sex increases the efficacy of natural selection in experimental yeast populations. AB - Why sex evolved and persists is a problem for evolutionary biology, because sex disrupts favourable gene combinations and requires an expenditure of time and energy. Further, in organisms with unequal-sized gametes, the female transmits her genes at only half the rate of an asexual equivalent (the twofold cost of sex). Many modern theories that provide an explanation for the advantage of sex incorporate an idea originally proposed by Weismann more than 100 years ago: sex allows natural selection to proceed more effectively because it increases genetic variation. Here we test this hypothesis, which still lacks robust empirical support, with the use of experiments on yeast populations. Capitalizing on recent advances in the molecular biology of recombination in yeast, we produced by genetic manipulation strains that differed only in their capacity for sexual reproduction. We show that, as predicted by the theory, sex increases the rate of adaptation to a new harsh environment but has no measurable effect on fitness in a new benign environment where there is little selection. PMID- 15800623 TI - Functional consequences of a CKIdelta mutation causing familial advanced sleep phase syndrome. AB - Familial advanced sleep phase syndrome (FASPS) is a human behavioural phenotype characterized by early sleep times and early-morning awakening. It was the first human, mendelian circadian rhythm variant to be well-characterized, and was shown to result from a mutation in a phosphorylation site within the casein kinase I (CKI)-binding domain of the human PER2 gene. To gain a deeper understanding of the mechanisms of circadian rhythm regulation in humans, we set out to identify mutations in human subjects leading to FASPS. We report here the identification of a missense mutation (T44A) in the human CKIdelta gene, which results in FASPS. This mutant kinase has decreased enzymatic activity in vitro. Transgenic Drosophila carrying the human CKIdelta-T44A gene showed a phenotype with lengthened circadian period. In contrast, transgenic mice carrying the same mutation have a shorter circadian period, a phenotype mimicking human FASPS. These results show that CKIdelta is a central component in the mammalian clock, and suggest that mammalian and fly clocks might have different regulatory mechanisms despite the highly conserved nature of their individual components. PMID- 15800624 TI - Deletion of active ADAMTS5 prevents cartilage degradation in a murine model of osteoarthritis. AB - Human osteoarthritis is a progressive disease of the joints characterized by degradation of articular cartilage. Although disease initiation may be multifactorial, the cartilage destruction appears to be a result of uncontrolled proteolytic extracellular matrix destruction. A major component of the cartilage extracellular matrix is aggrecan, a proteoglycan that imparts compressive resistance to the tissue. Aggrecan is cleaved at a specific 'aggrecanase' site in human osteoarthritic cartilage; this cleavage can be performed by several members of ADAMTS family of metalloproteases. The relative contribution of individual ADAMTS proteases to cartilage destruction during osteoarthritis has not been resolved. Here we describe experiments with a genetically modified mouse in which the catalytic domain of ADAMTS5 (aggrecanase-2) was deleted. After surgically induced joint instability, there was significant reduction in the severity of cartilage destruction in the ADAMTS5 knockout mice compared with wild-type mice. This is the first report of a single gene deletion capable of abrogating the course of cartilage destruction in an animal model of osteoarthritis. These results demonstrate that ADAMTS5 is the primary 'aggrecanase' responsible for aggrecan degradation in a murine model of osteoarthritis, and suggest rational strategies for therapeutic intervention in osteoarthritis. PMID- 15800625 TI - ADAMTS5 is the major aggrecanase in mouse cartilage in vivo and in vitro. AB - Aggrecan is the major proteoglycan in cartilage, endowing this tissue with the unique capacity to bear load and resist compression. In arthritic cartilage, aggrecan is degraded by one or more 'aggrecanases' from the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family of proteinases. ADAMTS1, 8 and 9 have weak aggrecan-degrading activity. However, they are not thought to be the primary aggrecanases because ADAMTS1 null mice are not protected from experimental arthritis, and cleavage by ADAMTS8 and 9 is highly inefficient. Although ADAMTS4 and 5 are expressed in joint tissues, and are known to be efficient aggrecanases in vitro, the exact contribution of these two enzymes to cartilage pathology is unknown. Here we show that ADAMTS5 is the major aggrecanase in mouse cartilage, both in vitro and in a mouse model of inflammatory arthritis. Our data suggest that ADAMTS5 may be a suitable target for the development of new drugs designed to inhibit cartilage destruction in arthritis, although further work will be required to determine whether ADAMTS5 is also the major aggrecanase in human arthritis. PMID- 15800626 TI - Cyclophilin D-dependent mitochondrial permeability transition regulates some necrotic but not apoptotic cell death. AB - Mitochondria play an important role in energy production, Ca2+ homeostasis and cell death. In recent years, the role of the mitochondria in apoptotic and necrotic cell death has attracted much attention. In apoptosis and necrosis, the mitochondrial permeability transition (mPT), which leads to disruption of the mitochondrial membranes and mitochondrial dysfunction, is considered to be one of the key events, although its exact role in cell death remains elusive. We therefore created mice lacking cyclophilin D (CypD), a protein considered to be involved in the mPT, to analyse its role in cell death. CypD-deficient mice were developmentally normal and showed no apparent anomalies, but CypD-deficient mitochondria did not undergo the cyclosporin A-sensitive mPT. CypD-deficient cells died normally in response to various apoptotic stimuli, but showed resistance to necrotic cell death induced by reactive oxygen species and Ca2+ overload. In addition, CypD-deficient mice showed a high level of resistance to ischaemia/reperfusion-induced cardiac injury. Our results indicate that the CypD dependent mPT regulates some forms of necrotic death, but not apoptotic death. PMID- 15800627 TI - Loss of cyclophilin D reveals a critical role for mitochondrial permeability transition in cell death. AB - Mitochondria play a critical role in mediating both apoptotic and necrotic cell death. The mitochondrial permeability transition (mPT) leads to mitochondrial swelling, outer membrane rupture and the release of apoptotic mediators. The mPT pore is thought to consist of the adenine nucleotide translocator, a voltage dependent anion channel, and cyclophilin D (the Ppif gene product), a prolyl isomerase located within the mitochondrial matrix. Here we generated mice lacking Ppif and mice overexpressing cyclophilin D in the heart. Ppif null mice are protected from ischaemia/reperfusion-induced cell death in vivo, whereas cyclophilin D-overexpressing mice show mitochondrial swelling and spontaneous cell death. Mitochondria isolated from the livers, hearts and brains of Ppif null mice are resistant to mitochondrial swelling and permeability transition in vitro. Moreover, primary hepatocytes and fibroblasts isolated from Ppif null mice are largely protected from Ca2+-overload and oxidative stress-induced cell death. However, Bcl-2 family member-induced cell death does not depend on cyclophilin D, and Ppif null fibroblasts are not protected from staurosporine or tumour-necrosis factor-alpha-induced death. Thus, cyclophilin D and the mitochondrial permeability transition are required for mediating Ca2+- and oxidative damage induced cell death, but not Bcl-2 family member-regulated death. PMID- 15800628 TI - Structural insights into mRNA recognition from a PIWI domain-siRNA guide complex. AB - RNA interference and related RNA silencing phenomena use short antisense guide RNA molecules to repress the expression of target genes. Argonaute proteins, containing amino-terminal PAZ (for PIWI/Argonaute/Zwille) domains and carboxy terminal PIWI domains, are core components of these mechanisms. Here we show the crystal structure of a Piwi protein from Archaeoglobus fulgidus (AfPiwi) in complex with a small interfering RNA (siRNA)-like duplex, which mimics the 5' end of a guide RNA strand bound to an overhanging target messenger RNA. The structure contains a highly conserved metal-binding site that anchors the 5' nucleotide of the guide RNA. The first base pair of the duplex is unwound, separating the 5' nucleotide of the guide from the complementary nucleotide on the target strand, which exits with the 3' overhang through a short channel. The remaining base paired nucleotides assume an A-form helix, accommodated within a channel in the PIWI domain, which can be extended to place the scissile phosphate of the target strand adjacent to the putative slicer catalytic site. This study provides insights into mechanisms of target mRNA recognition and cleavage by an Argonaute siRNA guide complex. PMID- 15800629 TI - Structural basis for 5'-end-specific recognition of guide RNA by the A. fulgidus Piwi protein. AB - RNA interference (RNAi) is a conserved sequence-specific gene regulatory mechanism mediated by the RNA-induced silencing complex (RISC), which is composed of a single-stranded guide RNA and an Argonaute protein. The PIWI domain, a highly conserved motif within Argonaute, has been shown to adopt an RNase H fold critical for the endonuclease cleavage activity of RISC. Here we report the crystal structure of Archaeoglobus fulgidus Piwi protein bound to double-stranded RNA, thereby identifying the binding pocket for guide-strand 5'-end recognition and providing insight into guide-strand-mediated messenger RNA target recognition. The phosphorylated 5' end of the guide RNA is anchored within a highly conserved basic pocket, supplemented by the carboxy-terminal carboxylate and a bound divalent cation. The first nucleotide from the 5' end of the guide RNA is unpaired and stacks over a conserved tyrosine residue, whereas successive nucleotides form a four-base-pair RNA duplex. Mutation of the corresponding amino acids that contact the 5' phosphate in human Ago2 resulted in attenuated mRNA cleavage activity. Our structure of the Piwi-RNA complex, and that determined elsewhere, provide direct support for the 5' region of the guide RNA serving as a nucleation site for pairing with target mRNA and for a fixed distance separating the RISC-mediated mRNA cleavage site from the anchored 5' end of the guide RNA. PMID- 15800630 TI - Friction and torque govern the relaxation of DNA supercoils by eukaryotic topoisomerase IB. AB - Topoisomerases relieve the torsional strain in DNA that is built up during replication and transcription. They are vital for cell proliferation and are a target for poisoning by anti-cancer drugs. Type IB topoisomerase (TopIB) forms a protein clamp around the DNA duplex and creates a transient nick that permits removal of supercoils. Using real-time single-molecule observation, we show that TopIB releases supercoils by a swivel mechanism that involves friction between the rotating DNA and the enzyme cavity: that is, the DNA does not freely rotate. Unlike a nicking enzyme, TopIB does not release all the supercoils at once, but it typically does so in multiple steps. The number of supercoils removed per step follows an exponential distribution. The enzyme is found to be torque-sensitive, as the mean number of supercoils per step increases with the torque stored in the DNA. We propose a model for topoisomerization in which the torque drives the DNA rotation over a rugged periodic energy landscape in which the topoisomerase has a small but quantifiable probability to religate the DNA once per turn. PMID- 15800631 TI - Sizing up the odds. PMID- 15800632 TI - Ontario. PMID- 15800634 TI - Recruiters and industry. Job creation and environmental protection. PMID- 15800638 TI - A feat worth replicating. PMID- 15800637 TI - Purified Argonaute2 and an siRNA form recombinant human RISC. AB - Genetic, biochemical and structural studies have implicated Argonaute proteins as the catalytic core of the RNAi effector complex, RISC. Here we show that recombinant, human Argonaute2 can combine with a small interfering RNA (siRNA) to form minimal RISC that accurately cleaves substrate RNAs. Recombinant RISC shows many of the properties of RISC purified from human or Drosophila melanogaster cells but also has surprising features. It shows no stimulation by ATP, suggesting that factors promoting product release are missing from the recombinant enzyme. The active site is made up of a unique Asp-Asp-His (DDH) motif. In the RISC reconstitution system, the siRNA 5' phosphate is important for the stability and the fidelity of the complex but is not essential for the creation of an active enzyme. These studies demonstrate that Argonaute proteins catalyze mRNA cleavage within RISC and provide a source of recombinant enzyme for detailed biochemical studies of the RNAi effector complex. PMID- 15800639 TI - Editor in the field Princeton's Lewis-Sigler Institute. PMID- 15800640 TI - A manually curated functional annotation of the human X chromosome. PMID- 15800641 TI - Frequency of new copy number variation in humans. PMID- 15800643 TI - Partners in crime. PMID- 15800644 TI - Sirtuins for healthy neurons. PMID- 15800645 TI - Defining stroke risks in sickle cell anemia. PMID- 15800646 TI - The first steps in adaptive evolution. PMID- 15800647 TI - The X chromosome: not just her brother's keeper. PMID- 15800651 TI - 3',5'-cyclic nucleotide phosphodiesterase 11A: localization in human tissues. AB - 3',5'-Cyclic nucleotide phosphodiesterase 11 (PDE11) is the most recently discovered family of human 3',5'-cyclic nucleotide phosphodiesterases (PDEs). This family contains one gene, PDE11A, with four splice variants (PDE11A1 PDE11A4). The physiological role of PDE11A has not been determined. Tadalafil (Cialis), a PDE5A inhibitor used for the treatment of male erectile dysfunction, has been reported to partially inhibit PDE11. It was therefore of interest to consider the pattern of expression of PDE11 in human tissues. Although four PDE11A mRNA transcripts have been reported, we detected protein corresponding to only one of them, PDE11A4, in human prostate, pituitary, heart and liver. Using immunohistochemistry, there was strong PDE11A antibody staining in the glandular epithelium of the prostate and weak staining of neuronal cells within parasympathetic ganglia in the heart. No PDE11A protein was detected in blood vessels or cardiac myocytes. None of the four potential PDE11A proteins were detected in human skeletal muscle, testis, or penis. PMID- 15800652 TI - Assessment of sexual functions in patients with chronic liver disease. AB - We aimed to determine the sexual problems and the prevalence of erectile dysfunction (ED) in patients with chronic liver disease by means of International Index of Erectile Function (IIEF). A total of 81 patients with stable chronic liver disease were included in the study. Patients were grouped as mild to moderate (Child Pugh A and B) (n=10), chronic hepatitis (hepatitis B, C and D) (n=28) and carriers (n=43) according to the type of their chronic liver disease. All patients were asked to complete a questionnaire including IIEF and demographics. The IIEF domain scores were calculated and ED grading was determined on erectile function domain. IIEF domain scores were compared between these groups. The mean age was 54.8 +/- 10.8 y. Using the IIEF, the prevalence of any ED was 50.6% for all patients, and 50, 50, and 51.1% for cirrhosis, chronic hepatitis and carriers, respectively. The IIEF domain scores were not significantly different among the patient groups. Sexual functions did not appear to be affected by the stable chronic liver disease. PMID- 15800653 TI - Water-soluble lipopolymer as a gene carrier to corpus cavernosum. AB - Adenovirus or naked plasmid DNA (pDNA) has been used to deliver the therapeutic gene into corpus cavernosum. However, the potential risks of viral vector and inefficiency of naked pDNA have limited their clinical application. In this study, water-soluble lipopolymer (WSLP) was evaluated as a gene carrier to corpus cavernosum. The WSLP/pDNA complex was transfected to smooth muscle cells in vitro. WSLP had high transfection efficiency, which was comparable to poly(ethylenimine) (PEI). In addition, WSLP had much less cytotoxicity than PEI, suggesting that WSLP is a safer carrier than PEI. To evaluate the transfection efficiency to corpus cavernosum, the WSLP/pDNA complex was injected into the rat corpus cavernosum. As a result, the WSLP/pDNA complex showed higher transfection efficiency than naked pDNA. In addition, the gene expression was dependent upon the dose of the complex. The results suggest that WSLP may be useful for gene therapy of erectile dysfunction. PMID- 15800654 TI - Phosphodiesterase 11 (PDE11) regulation of spermatozoa physiology. AB - Fertilization is well correlated with sperm concentration, rate of forward motility, and percentage of live, uncapacitated ejaculated spermatozoa, which is regulated in part by cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). Phosphodiesterases (PDEs) hydrolyze cyclic nucleotides to their corresponding monophosphates, thereby counterbalancing the activities of cAMP and cGMP, and PDE11 is highly expressed in the testis, prostate, and developing spermatozoa. However, a physiological role of PDE11 is not known. We generated PDE11 knockout (PDE11-/-) mice to investigate the role of PDE11 in spermatozoa physiology. Ejaculated sperm from PDE11-/- mice displayed reduced sperm concentration, rate of forward progression, and percentage of live spermatozoa. Pre-ejaculated sperm from PDE11-/- mice displayed increased premature/spontaneous capacitance. These data are consistent with human data and suggest a role for PDE11 in spermatogenesis and fertilization potential. This is the first phenotype described for the PDE11-/- mouse and the first report of a physiological role for PDE11. PMID- 15800657 TI - Detection of plasmid DNA vectors following gene transfer to the murine airways. AB - Non-viral gene therapy is being considered as a treatment for cystic fibrosis. In clinical studies and in studies using the mouse airways as a model, current formulations result in only transient transgene expression. A number of reasons for this have been proposed including the loss of plasmid DNA from cells. The aim of these studies was to investigate why transgene expression from non-viral vectors is transient in the mouse lung. Plasmid DNA encoding the luciferase reporter gene was complexed with the cationic lipid GL67 and delivered to the mouse airways. The persistence of plasmid DNA in the mouse lungs was investigated using quantitative PCR and Southern hybridization. Results showed that intact plasmid DNA persisted in the mouse lung in the absence of any detectable luciferase activity. The de novo methylation of plasmid DNA in vivo was investigated as a potential cause of this transient gene expression but results suggested that plasmid DNA does not become de novo methylated in the mouse lung. Therefore processes other than the loss of plasmid DNA from the lung or the de novo methylation of plasmid DNA vectors must be responsible for the transient transgene expression. PMID- 15800658 TI - Combination of gemcitabine and Ad5/3-Delta24, a tropism modified conditionally replicating adenovirus, for the treatment of ovarian cancer. AB - Conditionally replicating adenoviruses (CRAds) represent a novel approach for the treatment of cancers resistant to conventional therapies. The efficacy of CRAds might be further improved by using chemotherapeutic agents in a multimodal antitumor approach. We have evaluated the use of Ad5/3-Delta24, a serotype 3 receptor targeted Rb/p16 pathway selective CRAd, in combination with gemcitabine against human ovarian adenocarcinoma. The combination of these agents showed synergistic cell killing in vitro compared to single treatments. However, the effect was dependent on dose and sequencing of the agents. Our results also indicate that gemcitabine reduces the initial rate of Ad5/3-Delta24 replication without affecting the total amount of virus produced. Possible reasons for synergy between Ad5/3-Delta24 and gemcitabine include the chemosensitizing activity of E1A and/or altered replication kinetics. In an orthotopic murine model of peritoneally disseminated ovarian cancer, the combination increased the survival of mice over either agent alone, and almost 60% of treated mice were cured. Sequencing of the agents was critical for toxicity versus efficacy. Mice remained free from intraperitoneal disease, but some succumbed to treatment related hepatic or bone marrow toxicity. This suggests that improved efficacy may uncover treatment-related toxicity, which needs to be monitored closely in clinical trials. PMID- 15800656 TI - Cancer immunotherapy using a DNA vaccine encoding a single-chain trimer of MHC class I linked to an HPV-16 E6 immunodominant CTL epitope. AB - The potency of DNA vaccines may be affected by the efficiency of intracellular processing and MHC class I presentation of encoded antigens. Since a single-chain trimer (SCT) composed of peptide, beta2-microglobulin (beta2m), and MHC class I heavy chain has been shown to bypass antigen processing and lead to stable presentation of peptides, we investigated the efficacy of a DNA vaccine encoding a SCT composed of an immunodominant CTL epitope of human papillomavirus type 16 (HPV-16) E6 antigen, beta2m, and H-2Kb MHC class I heavy chain (pIRES-E6-beta2m Kb). Transfection of 293 cells with pIRES-E6-beta2m-Kb can bypass antigen processing and lead to stable presentation of E6 peptide. Furthermore, C57BL/6 mice vaccinated with pIRES-E6-beta2m-Kb exhibited significantly increased E6 peptide-specific CD8+ T-cell immune responses compared to mice vaccinated with DNA encoding wild-type E6. Most importantly, 100% of mice vaccinated with pIRES E6-beta2m-Kb DNA were protected against a lethal challenge of E6-expressing TC-1 tumor cells. In contrast, all mice vaccinated with wild-type E6 DNA or control plasmid DNA grew tumors. Our data indicate that a DNA vaccine encoding a SCT can lead to stable enhanced MHC class I presentation of encoded antigenic peptide and may be useful for improving DNA vaccine potency to control tumors or infectious diseases. PMID- 15800659 TI - A hypoxia-inducible vigilant vector system for activating therapeutic genes in ischemia. AB - Hypoxia represents an endogenous pathophysiological signal underlying cell growth, adaptation and death in a variety of diseases, including ischemic heart diseases, stroke and solid tumors. A vigilant vector system depends on a gene switch which can sense the hypoxia signal occurring in ischemic events and turn on/off protective gene expressions when necessary. This system uses the oxygen dependent degradation domain derived from hypoxia-inducible factor 1alpha as the hypoxia sensor and a double-vector system as signal amplifier. For treating ischemic heart diseases, a cardiac-specific MLC-2v promoter is used to deliver transgenes specifically to the heart. When tested in cardiomyocyte cultures, it produced a rapid and robust gene induction upon exposure to low oxygen. In a mouse model for myocardial infarction, the vigilant vectors turned on therapeutic genes such as heme oxygenase-1 in response to ischemia, significantly reduced apoptosis in the infarct area and improved cardiac functions. The hypoxia regulated gene transfer afforded by the vigilant vectors may provide a powerful tool for delivering therapeutic proteins specifically to ischemic tissues with optimal physiological control. PMID- 15800660 TI - Inhibition of NF-kappaB enhances the cytotoxicity of virus-directed enzyme prodrug therapy and oncolytic adenovirus cancer gene therapy. AB - Virus-directed enzyme prodrug therapy utilizing the bacterial enzyme nitroreductase delivered by a replication-defective adenovirus vector to activate the prodrug CB1954 is a promising strategy currently undergoing clinical trials in patients with a range of cancers. Similarly, selectively replicating oncolytic adenoviruses are entering clinical trials. An understanding of interactions between vector and target cell are critical to the development of these strategies. We demonstrate that adenovirus vectors activate cellular pathways that promote cell survival in an NF-kappaB-dependent manner, and consequently have a negative effect on the efficacy of cell killing induced by cancer gene therapy strategies. This provides a potential therapeutic target to enhance the cytotoxicity of these approaches. PMID- 15800661 TI - Transcriptional targeting of acute hypoxia in the tumour stroma is a novel and viable strategy for cancer gene therapy. AB - Deregulated tumour growth and neovascularization result in an inadequate tumour blood supply, leading to areas of chronic hypoxia and necrosis. Irregular vascular structure and abnormal tumour physiology also cause erratic blood flow in tumour vessels. We reasoned that tumour stroma, including vascular endothelial cells, would consequently experience transient hypoxia that may allow transcriptional targeting as part of an antivascular gene therapy approach to cancer. To exploit hypoxia for transcriptional regulation, retroviral vectors were generated with modified LTRs: a 6-mer of hypoxia response elements in place of the viral enhancer produced near wild-type levels of expression in hypoxia but was functionally inert in normoxia. In a tumour xenograft model, expression was mainly around areas of necrosis, which were shown to be hypoxic; no expression was detected in tumour stroma. Time-course experiments in vitro demonstrated that expression was transient in response to a hypoxic episode, such that a reporter gene would be insensitive to acute hypoxia in vivo. In contrast, a significant therapeutic effect was seen upon ganciclovir administration with a vector expressing thymidine kinase (TK) in the tumour stroma. Expression of TK was more effective when targeted to acute hypoxia in the stroma compared to chronic hypoxia in the poorly vascularized regions of the tumour cell compartment. The data presented here are evidence that hypoxia in the stromal compartment does occur and that transient hypoxia constitutes a valid therapeutic target. PMID- 15800662 TI - Inhibition of ets, an essential transcription factor for angiogenesis, to prevent the development of abdominal aortic aneurysm in a rat model. AB - The pathophysiology of abdominal aortic aneurysms (AAA) is considered to be complicated. As matrix degradation contributes to the progression of AAA, the destruction and degradation of elastin fibers caused by an increase in matrix metalloproteinases (MMPs) plays a pivotal role in the development of AAA. Although ets, an essential transcription factor for angiogenesis, regulates MMPs, the role of ets in the development of AAA has not yet been clarified. Thus, we evaluated the role of ets in a rat AAA model using a decoy strategy. Transfection of ODN into AAA was performed by transient aortic perfusion of elastase and by wrapping the AAA in a delivery sheet containing decoy ODN. The inhibitory effect of ets decoy ODN on ets binding activity was confirmed by gel mobility shift assay. MMPs expression was decreased in the aorta transfected with ets decoy ODN as compared to scrambled decoy ODN. Also, ultrasound study demonstrated that elastase-induced aneurismal dilation was significantly suppressed by transfection of ets decoy ODN at 4 weeks after treatment as compared to scrambled decoy ODN. Moreover, the destruction of elastin fibers was inhibited in the aorta transfected with ets decoy ODN, accompanied by a reduction of MMPs expression. An inhibitory effect of decoy ODN on MMP expression was confirmed by ex vivo experiments showing that transfection of decoy ODN into an organ culture of human aorta resulted in significant inhibition of the secretion of both MMP-1 and MMP 9. Here, we demonstrated that ets may play a pivotal role in the progression of AAA through the activation of MMPs in a rat model. Ets might be a potential target to develop pharmacotherapy/gene therapy to treat AAA through the inhibition of MMPs. PMID- 15800663 TI - A novel DNA vaccine based on ubiquitin-proteasome pathway targeting 'self' antigens expressed in melanoma/melanocyte. AB - Cancer vaccine that targets 'self'-antigens expressed at high levels in tumor cells is a potentially useful immunotherapy, but immunological tolerance often defeats this strategy. Here, we describe the use of a naked DNA vaccine encoding a self tumor antigen, tyrosinase-related protein 2, to whose N-terminus ubiquitin is fused in a 'nonremovable' fashion. Unlike conventional DNA vaccines, this vaccine broke the tolerance and induced protective immunity to melanoma in C57BL/6 mice, as evaluated by tumor growth, survival rate and lung metastasis. The protective immunity was cancelled in the proteasome activator PA28alpha/beta knockout mice. Moreover, this vaccination exhibited therapeutic effects on melanoma implanted before vaccination. Our findings provide evidence for the first time that naked DNA vaccines encoding a ubiquitin-fused self-antigen preferentially induce the main effector CD8+ T cells through efficient proteolysis mediated by the ubiquitin-proteasome pathway, and lead the way to strategies aimed at targeting tissue differentiation antigens expressed by tumors. PMID- 15800664 TI - The cost-effectiveness of candesartan-based antihypertensive treatment for the prevention of nonfatal stroke: results from the Study on COgnition and Prognosis in the Elderly. AB - Patients who survive a first stroke are often left with permanent disabilities, and have significant needs for rehabilitation and long-term care. Antihypertensive treatment reduces the risk of cardiovascular events such as stroke. The purpose of this study was to investigate the cost-effectiveness of candesartan-based antihypertensive treatment for the prevention of nonfatal stroke. The cost-effectiveness analysis was based on data from Study on COgnition and Prognosis in the Elderly (SCOPE), where patients were randomly assigned to receive the angiotensin receptor blocker candesartan or placebo, with open-label active antihypertensive treatment added as needed. The analysis was carried out using a Markov model, which combined clinical and resource utilization data from SCOPE with Swedish retail prices for drugs and unit costs for in-patient stays, and outpatient visits. The cost per patient was 1949 EUR in the candesartan group and 1578 EUR in the control group. The largest share of the cost was attributed to antihypertensive treatment in the candesartan group and to the long-term cost of stroke in the control group. Candesartan-based antihypertensive treatment was associated with 0.0289 additional quality-adjusted life-years (QALYs) per patient and an incremental cost per QALY gained of approximately 13,000 EUR. Sensitivity analyses showed that these results were fairly stable. In conclusion, the cost per QALY gained with candesartan-based antihypertensive treatment lies within the range of society's willingness to pay for health gains. The results indicate that candesartan-based antihypertensive treatment is cost-effective for the prevention of nonfatal stroke. PMID- 15800665 TI - Sleep-disordered breathing and hypertension among African Americans. AB - This study investigated differences in sleep-disordered breathing (SDB) between hypertensives without a family history of hypertension and hypertensives with a family history. Furthermore, it examined whether these two groups differed in the severity of SDB. Patients were African Americans (n=162, mean age=51.19+/-13.77 years; mean body mass index (BMI)=37.85+/-9.51 kg/m2, male=57%), who were referred to the clinic because of a sleep complaint. Sleep was recorded in the laboratory using standard physiological parameters; all parameters were analysed by a trained scorer. Altogether, 91% of the patients received an SDB diagnosis. Of these patients, 25% were hypertensives without a family history, 20% were hypertensives with a family history, and 55% were normotensives. We found a significant difference between these patient groups regarding the severity of SDB (F14,158=1.823, P<0.05), but no significant group difference was observed in the rate of SDB. Increasing weight was accompanied by increasing severity of SDB. The finding that hypertensive patients with or without a positive family history showed worse oxygenation and respiratory characteristics than did normotensives is consistent with previous research. Of note, hypertensives reporting a family history were characterized by a greater number of oxygen desaturations and apnoea hypopnoea index than those typified only by a current diagnosis of hypertension. Hypertensives with a family history are likely to show a profile of greater blood pressure, higher BMI, and more severe SDB, which by all accounts are more common among African Americans. PMID- 15800666 TI - Short intensive sequential therapy followed by autologous stem cell transplantation in adult Burkitt, Burkitt-like and lymphoblastic lymphoma. AB - The feasibility and efficacy of up-front high-dose sequential chemotherapy followed by autologous stem cell transplantation (ASCT) in previously untreated adults (median age 33 years; range 15-64) with Burkitt lymphoma (BL), Burkitt like lymphoma (BLL) or lymphoblastic lymphoma (LyLy), both without central nervous system or extensive bone marrow involvement was investigated in a multicenter phase II study. Treatment consisted of two sequential high-dose chemotherapy induction courses incorporating prednisone, cyclophosphamide, doxorubicin, etoposide and mitoxantrone, without high-dose methotrexate or high dose cytarabine. Patients with at least PR went on with BEAM and ASCT. Protocol treatment was completed by 23/27 (85%) BL/BLL and 13/15 (87%) LyLy patients. Median treatment duration until BEAM was 70 (range: 50-116) days. No toxic deaths occurred. Response to treatment was complete response (CR) 81% and partial response (PR) 11% for BL/BLL, CR 73% and PR 20% for LyLy. At a median follow-up of 61 months of patients still alive, six BL/BLL and eight LyLy patients have died. The actuarial 5-year overall and event-free survival estimates are 81 and 73% for BL/BLL vs 46 and 40% for LyLy patients. In conclusion, this short up front high-dose sequential chemotherapy regimen, followed by ASCT is highly effective in adults with BL/BLL with limited bone marrow involvement, but less so in patients with LyLy. PMID- 15800668 TI - G1 arrest by p16INK4A uncouples growth from cell cycle progression in leukemia cells with deregulated cyclin E and c-Myc expression. AB - The cell cycle inhibitor p16(INK4A) is frequently inactivated in acute lymphoblastic T-cell leukemia (T-ALL). We analyzed mechanisms and consequences of p16(INK4A) reconstitution in T-ALL cells lacking this tumor suppressor. CCRF-CEM cells with tetracycline-regulated p16(INK4A) expression underwent stable G1-phase cell cycle arrest for 72 h followed by massive apoptosis. p16(INK4A) expression caused pRB hypophosphorylation and repression of certain E2F target genes. Interestingly, cyclin E and c-Myc were not affected, suggesting pRB/E2F independent expression of these E2F targets. Cyclin E/CDK2, however, was inactive due to stabilization and redistribution of p27(Kip1) from CDK4/CDK6 to CDK2. Analyses of c-Myc target genes suggested that c-Myc was transcriptionally inactive, which correlated with hypophosphorylation of the c-Myc inhibitor p107. Thus, p16(INK4A), although unable to repress the expression of deregulated cyclin E and c-Myc, functionally inactivated these potential oncogenes. p16(INK4A) arrested cells showed morphologic changes, induction of T-cell-specific surface markers and repression of telomerase activity, suggesting differentiation. Moreover, p16(INK4A) reconstitution was associated with increased cellular volume, normal protein synthesis rates and elevated ATP levels. Taken together, p16(INK4A) reconstitution in p16(INK4A)-deficient T-ALL cells induced cell cycle arrest in the presence of cyclin E and c-Myc expression, uncoupled growth from cell cycle progression and caused a sequential process of growth, differentiation and apoptosis. PMID- 15800669 TI - Five decades of therapy for multiple myeloma: a paradigm for therapeutic models. PMID- 15800667 TI - Hematopoietic cell transplantation from HLA-identical sibling donors after low dose radiation-based conditioning for treatment of CML. AB - A total of 24 patients (median age 58; range, 27-71 years) with chronic myeloid leukemia (CML) in first chronic (CP1) (n=14), second chronic (n=4), or accelerated phase (n=6) who were not candidates for conventional hematopoietic cell transplantation (HCT), received nonmyeloablative HCT from HLA-matched siblings a median of 28.5 (range, 11-271) months after diagnosis. They were conditioned with 2 Gy total body irradiation (TBI) alone (n=8) or combined with fludarabine, 90 mg/m(2) (n=16). Postgrafting immunosuppression included cyclosporine and mycophenolate mofetil. All patients initially engrafted. However, 4 of 8 patients not given fludarabine experienced nonfatal rejection while all others had sustained engraftment. With a median follow-up of 36 (range, 4-49) months, 13 of 24 patients (54%) were alive and in complete remission. There were five (21%) deaths from nonrelapse mortality, one (4%) during the first 100 days after transplant. The proportions of grade II, III, and IV acute GVHD were 38, 4, and 8%, respectively. The 2-year estimate of chronic GVHD was 32%. The 2 year survival estimates for patients in CP1 (n=14) and beyond CP1 (n=10) were 70 and 56%, respectively. This study shows encouraging remission rates for patients with CML not eligible for conventional allografting. PMID- 15800670 TI - CLL family 'Pedigree 14' revisited: 1947-2004. AB - The notion that inherited predisposition contributes to the development of haematological malignancies is generally thought of as being a relatively new idea. However, Videbaek made a clear enunciation of such a hypothesis in 1947, from a study of tumour incidence in relatives of patients with different leukaemias. To gain further insight into inherited susceptibility to chronic lymphocytic leukaemia (CLL), we followed up the descendants of Videbaek's 'Pedigree 14' series of families. Using the Danish medical and pedigree databases, complete tracing of 222 descendants of the original 57 family members was achieved. To date, 10 family members have been diagnosed with CLL, one with T cell lymphoma and 17 with nonhaematological cancers, including five with breast cancer. The detailed follow up of this family provides further support for inherited predisposition to CLL and illustrates the value of follow-up studies of previously published family material for genetic analyses. PMID- 15800671 TI - ZAP-70 is expressed by a subset of normal human B-lymphocytes displaying an activated phenotype. AB - The Syk family tyrosine kinase ZAP-70 is essential for normal T-cell development and signaling. Recently, leukemic cells from some patients with B-cell chronic lymphocytic leukemia (B-CLL) were shown to express ZAP-70. Owing to the prognostic value of B-CLL ZAP-70 expression, this phenotype may reflect intrinsic biological differences between the two subsets of disease. However, it remains unclear whether CLL-B cells aberrantly acquire ZAP-70 expression during the transformation process or whether ZAP-70 may be expressed under certain conditions in normal human B-lymphocytes. To discriminate between these two possibilities, we assessed ZAP-70 expression in normal human B-lymphocytes. Our data demonstrate that ZAP-70 is expressed in a subpopulation of tonsillar and splenic normal B-lymphocytes that express an activated phenotype. Furthermore, ZAP-70 expression can be induced in vitro upon stimulation of blood and tonsillar B cells. Finally, we show that phosphorylation of ZAP-70 occurs in tonsillar B cells with stimulation through the B-cell receptor. These results provide new insight into normal human B-cell biology as well as provide clues about the transformed cell in B-CLL. PMID- 15800672 TI - Donor leukocyte infusion after hematopoietic stem cell transplantation in patients with juvenile myelomonocytic leukemia. AB - Juvenile myelomonocytic leukemia (JMML) is a clonal myeloproliferative disorder of early childhood. In all, 21 patients with JMML who received donor leukocyte infusion (DLI) after allogeneic hematopoietic stem cell transplantation (HSCT) for either mixed chimerism (MC, n=7) or relapse (n=14) were studied. Six patients had been transplanted from an HLA-matched sibling and 15 from other donors. Six of the 21 patients (MC: 3/7 patients; relapse: 3/14 patients) responded to DLI. Response rate was significantly higher in patients receiving a higher total T cell dose (> or =1 x 10(7)/kg) and in patients with an abnormal karyotype. None of the six patients receiving DLI from a matched sibling responded. Response was observed in five of six patients who did and in one of 15 children who did not develop acute graft-versus-host disease following DLI (P=0.01). The overall outcome was poor even for the responders. Only one of the responders is alive in remission, two relapsed, and three died of complications. In conclusion, this study shows that some cases of JMML may be sensitive to DLI, this providing evidence for a graft-versus-leukemia effect in JMML. Infusion of a high number of T cells, strategies to reduce toxicity, and cytoreduction prior to DLI may improve the results. PMID- 15800673 TI - The t(8;17)(p11;q23) in the 8p11 myeloproliferative syndrome fuses MYO18A to FGFR1. AB - The 8p11 myeloproliferative syndrome (EMS) also known as stem cell leukemia lymphoma syndrome (SCLL) is associated with translocations that disrupt FGFR1. The resultant fusion proteins are constitutively active tyrosine kinases, and different FGFR1 fusions are associated with subtly different disease phenotypes. We report here a patient with a t(8;17)(p11;q23) and an unusual myelodysplastic/myeloproliferative disease (MDS/MPD) characterized by thrombocytopenia due to markedly reduced size and numbers of megakaryocytes, with elevated numbers of monocytes, eosinophils and basophils. A novel mRNA fusion between exon 32 of the myosin XVIIIA gene (MYO18A) at chromosome band 17q11 and exon 9 of FGFR1 was identified. Partial characterization of the genomic breakpoints in combination of bubble-PCR with fluorescence in situ hybridization revealed that the t(8;17) arose from a three-way translocation with breaks at 8p11, 17q11 and 17q23. MYO18A-FGFR1 is structurally similar to other fusion tyrosine kinases and is likely to be the causative transforming lesion in this unusual MDS/MPD. PMID- 15800674 TI - Can application of serine protease inhibitors TPCK and TLCK provide evidence for possible involvement of serine protease Omi/HtrA2 in imatinib mesylate-induced cell death of BCR-ABL-positive human leukemia cells? PMID- 15800675 TI - Angiogenesis in Hodgkin's lymphoma: a morphometric approach in 286 patients with prognostic implications. AB - The significance of angiogenesis in Hodgkin's lymphoma (HL) is not well defined. The aim of this study was to evaluate various morphometric characteristics of microvessels in lymph node sections of 286 patients with HL at diagnosis and investigate their relationship with clinicopathologic parameters and prognosis. Microvessel density (MVD), total vascular area (TVA) and several size- and shape related microvascular parameters were quantitated--after anti-CD34 immunohistochemical staining--in the region of most intense vascularization, using image analysis. An increase in microvessel caliber parameters (area, perimeter, major and minor axis length) and a decrease in MVD were noted with increasing stage. An inverse relationship was recorded between MVD and the number of involved sites (NIS) and LDH. In univariate analysis, overall disease-specific survival was adversely affected by MVD and TVA, whereas inferior failure-free survival (FFS) was associated with the presence of more flattened vessel sections. Multivariate analysis disclosed that the extent of angiogenesis (MVD/TVA), age and the NIS independently affected overall survival. Accordingly, FFS was independently linked to the shape of microvessels and albumin levels or the NIS. In conclusion, our data support the view that angiogenesis in HL provides independent prognostic information, requiring the concomitant evaluation of quantitative and qualitative aspects of microvascular network. PMID- 15800676 TI - Genomics of lung cancer may change diagnosis, prognosis and therapy. AB - Despite significant improvements in tumor management in general, the prognosis of lung cancer patients remains dismal. It is a hope that our increasing knowledge in molecular aspects of tumor development, growth and progression will open new targets for therapeutic interventions. In this review we discuss some of the more recent results of this field. This includes the susceptibility factors, an association between genetic changes in EGFR pathway and tyrosine kinase inhibitors, the role of gene hypermethylation and genetic profiling, as well as different molecular aspects of tumor progression. Available data all support that lung cancer is a group of diseases with not only distinct histological but with similarly different genetic characters. Accordingly, the diagnosis, prognosis and therapy must accommodate this heterogeneity. PMID- 15800677 TI - Gastrointestinal stromal tumors: a clinicopathologic and immunohistochemical study of 136 cases. AB - The clinicopathologic features of 136 gastrointestinal stromal tumors were analyzed. The tumors occurred in 60 women and 76 men, ranging in age from 19 to 88 years (median 59 years, mean 59.2 years). Sixty-one cases arose from stomach, 38 from small intestine and 11 from colon or rectum. Abdominal cavity was indicated as tumor site in 10 cases, but the extra-gastrointestinal origin using strict criteria was not proved. Four locally recurrent cases and 12 metastatic samples were also included. The primary and recurrent tumors ranged in size from 0.5 to 30 cm (mean 8.3 cm). The large number of high-grade cases (85 of 112 classifiable) is alarming and emphasize the importance of oncology care. Histologically, ninety-two cases were classified as spindle cell while 11 as epithelioid GIST. Mixed cellularity was seen in 33 cases. Skeinoid fibers were present in 14 and coagulation necrosis in 40 primary cases. Ulceration observed by microscopic examination was common (36 of 110 cases, 32.7%), explaining the clinically frequently observed gastrointestinal bleeding. Unusual histological features such as stromal hyalinization and nuclear palisading were present in 30 and 27 cases, respectively. Immunohistochemical CD117 (c-kit) positivity was documented in 133 cases. Three cases with CD117 negative results were included, because their morphology was most consistent with GIST and immunohistochemical reactions excluded the possibility of other neoplasms. CD34 positivity was seen in 70%, alpha-smooth muscle actin positivity in 39.6% of examined cases. Only one case showed desmin reactivity and seven had S100 positive tumor cells. For h caldesmon 39 cases proved to be positive (60.9% of the tested cases). PMID- 15800678 TI - Polymorphisms of the XRCC1 DNA repair gene in head and neck cancer. AB - Inherited polymorphisms in the genes controlling the cell cycle or functioning in the DNA repair mechanisms may impair their function and contribute to genetic susceptibility. Abnormalities in the DNA repair have been reported in head and neck cancer. The XRCC1 gene functions in singlestrand break and base excision repair processes. In this study, two polymorphisms of the XRCC1 gene, Arg194Trp and Arg399Gln were investigated in 95 patients with head and neck carcinoma. The polymorphic regions were amplified by PCR followed by digestion with methylation specific restriction enzymes, and analyzed electrophoretically. Genotype and allele frequencies were calculated, and association with cancer risk or clinical parameters was investigated. No association was observed between the genotypes and head and neck cancer for either polymorphism. Distribution of the alleles did not significantly differ between the patients and the control group. A significant association was only found for the Trp194 allele among the smoking individuals. Our data indicate that the Arg194Trp and Arg399Gln polymorphisms do not confer a significant risk for head and neck carcinogenesis. PMID- 15800679 TI - Changes of cell adhesion and extracellular matrix (ECM) components in cervical intraepithelial neoplasia. AB - Cell-cell and cell-extracellular matrix interaction is crucial in tumor progression. Tight junction (TJ) proteins as occludin and claudins (CLDNs) play important role in this process together with several extracellular matrix components, as syndecan. Our previous work suggested significant changes in the expression of claudins even in the early stages of cervical carcinogenesis. The aim of our present work was to study the expression of occludin and syndecan-1, as compared to CLDNs, in early phases of cervical carcinogenesis. Paraffin sections of 50 samples were studied by immunohistochemistry, including cervical intraepithelial neoplasias (CINI-II-III), in situ carcinomas (CIS) and normal cervical samples. Occludin and CLDN-2 were found colocalized in the basal layer, while syndecan-1 and CLDN-1, -4 and -7 were coexpressed in the parabasal and intermedier layers in normal epithelia. Intensity of occludin staining decreased in CIN/CIS lesions, although it was more extended towards the upper epithelial layers with inverse relation with grades, as seen in the case of CLDN-2 expression. CLDN-1, -2, -4 and -7 were detected in the entire epithelium in CIN, showing decrease in CIS. The progression of CIN was associated with reduced syndecan-1 expression, in contrast to CLDN-1, -4 and -7 which increased toward CIS. The obtained data suggest that significant changes occur in the composition of cell adhesion complexes even in early stages of cervical carcinogenesis. The pattern of expression is characteristic for the alteration, the changes in the different components, however, are not parallel with each other. PMID- 15800680 TI - Immunohistochemical analysis of c-myc, c-jun and estrogen receptor in normal, hyperplastic and neoplastic endometrium. AB - To evaluate the role of c-jun and c-myc proto-oncogenes in normal, hyperplastic and neoplastic endometrium in relation to estrogen receptor (ER) status and to investigate whether these genes can be related to other histopathological features of endometrial carcinoma, 32 endometrial carcinomas, 38 endometrial hyperplasias and 22 cyclic endometria (10 proliferative and 12 secretory) were evaluated histologically. Endometrial hyperplasia cases were classified as simple and complex hyperplasia without atypia, and atypical hyperplasia. Endometrial carcinoma cases were subtyped according to the International Society of Gynecological Pathologists. Modified FIGO system was used for both grading and staging. Immunohistochemical examination was performed using antibodies to ER alpha, c-myc and c-jun with streptavidin-biotin-peroxidase technique. The mean percentage of ER-alpha positive cells changed cyclically during the menstrual cycle, and it was the highest (96%) and the lowest (31.6%) in proliferative and carcinomatous endometrium, respectively. There was a statistically significant difference between proliferative and secretory phases and proliferative and carcinomatous endometrium in relation to ER-alpha staining (p<0.05). There was also a statistically significant difference with respect to ERalpha reactivity between secretory phase and each hyperplastic group, as well as between the carcinoma group and each hyperplastic group (p<0.05). Although not significant, the mean percentage of c-myc expressing cells in the carcinoma group was higher (15.3%) than that of proliferative phase and hyperplastic groups. The mean percentage of c-jun positive cells in proliferative endometrium was slightly higher than in secretory endometrium, and it was the highest in atypical hyperplastic endometrium (28.3%), but there was no statistically significant difference between the groups. In carcinoma cases, a positive correlation was observed between c-jun positivity and tumor grade (p=0.027, r=0.3908), but such a correlation with c-myc was not found. A positive correlation was detected between ER-alpha and c-myc expression (p=0.038, r=0.3686). A progressive loss of ER seems to be correlated with increasing malignant transformation. C-myc expression might play a role in the development of endometrial carcinoma via ER. The association between c-jun and ER appears to be lost in endometrial carcinoma. The relationship between c-myc, c-jun and ER appears to be altered in endometrial carcinoma compared to that of menstrual endometrium. PMID- 15800681 TI - Is quadrant biopsy sufficient in men likely to have advanced prostate cancer? Comparison with extended biopsy. AB - We hypothesized that quadrant prostate biopsy (QPB) provides sufficient first line pathological evaluation of patients with presumed advanced prostate cancer (PC). The aim of this study was to investigate whether the reduction of core number in first-line PB from 6-12 to 4 in patients with presumed advanced PC leads to loss of clinically relevant information. We retrospectively studied 113 men that underwent PB, classified in two groups: "H" (high) and "L" (low likelihood of having advanced PC), according to PSA, digital rectal and transrectal ultrasound findings. Pathological results of 6-12-core PB and QPB were retrospectively compared for the presence of malignancy, percentage of positive cores, Gleason score (GS), and the presence of high-grade prostatic intraepithelial neoplasia (HGPIN). PC detection rate was not impaired in group H but dropped significantly in group L, and the percentage of positive cores was not significantly changed in group H (p=0.39), but decreased in group L (p=0.04), due to sampling scheme reduction. No HGPIN was missed with QPB in group H, while 2 HGPINs were missed in group L. No significant change in GS in either group was observed (p=0.12, p=0.13) due to reduction to QPB. We conclude that in patients with presumed advanced PC, reduction of the number of cores in PB may be an acceptable diagnostic strategy, but further studies are needed to analyze the impact of PB scheme reduction on other relevant pathological information obtained from PB. PMID- 15800683 TI - ROC analysis as an additional method to characterize time to event data. AB - The receiver operating characteristic (ROC) is predominantly used to assess the discriminant power of diagnostic tests. The present paper proposes this method as an additional alternative to compare survival data of in oncology or related fields. Survival data of brain tumors were analysed with conventional Kaplan Meier method and ROC. The Area und the Curve (ROC-curve) gives additional illustrative information to distinguish between two therapeutic approaches and low or high grade brain tumors. PMID- 15800682 TI - Expression of p53, Ki-67 and Bcl-2 in parathyroid adenoma and residual normal tissue. AB - The aim of this study was to investigate the expression of Ki-67, bcl-2 and p53 in parathyroid adenomas and their residual rim of normal parathyroid tissue. Specimens from 26 parathyroid adenomas were studied by immunohistochemical analysis for Ki-67, bcl-2 and p53 expression. Positive findings were noted for p53 in 4 (15%) adenomas and none of the residual rims of normal parathyroid tissue (p = 0.055); for Ki-67 in 15 (56%) adenomas and none of the residual rims of normal parathyroid tissue (p = 0.00002); and for bcl-2 in 19 (73%) adenomas and 8 (31%) residual rims of normal parathyroid tissue (p < 0.01). The high rate of Ki-67 expression may indicate susceptibility of parathyroid adenomas to clonal proliferation. The weak immunoreactive expression of p53, combined with a relatively strong expression of bcl-2, may contribute to the characteristic slow progression of these tumors. PMID- 15800684 TI - Cystic thymoma. AB - Thymic cysts are rare lesions located along the anatomical course of the third pharyngeal pouch. While most of the cases represent congenital cysts, they may also be related to neoplasms. We report a case of a micronodular thymoma with lymphoid stroma, which was completely built of small cysts, discuss the pathologic features of this tumor type and review the etiology and other aspects of thymic cysts. PMID- 15800685 TI - Endoscopic manpower in Romania seems deficient: appropriate training is mandatory. PMID- 15800686 TI - Esomeprazole in the treatment of patients with heartburn and other upper gastrointestinal symptoms, referred to primary care -- results of the in-practice evaluation program in Romania. AB - BACKGROUND AND AIM: The real-life clinical setting has been known to provide, sometimes, results different from controlled study outcomes. IPEP (InPractice Evaluation Program) was designed to evaluate the efficacy of esomeprazole in providing symptom relief in patients with reflux disease who present to primary care physicians for heartburn and symptoms that are poorly controlled by other drugs, thus testing the efficacy of the drug in day-to-day practice. METHODS: 119 primary care physicians and 22 internal medicine specialists, widely distributed geographically, participated in the program. The frequency and severity of three symptoms (heartburn, acid regurgitation and upper abdominal pain) were assessed before and at the end of one month treatment with esomeprazole 40 mg once daily. RESULTS: Follow-up data from 747 patients enrolled in IPEP was available to evaluate the efficacy of the intervention. Over 80% of the enrolled patients reported a decrease in the frequency of symptoms to less than one episode per week for all the three major symptoms (heartburn, acid regurgitation and upper abdominal pain). Regarding the severity of symptoms, 50-60% of the patients reported total relief, while 30-40% reported persistence of symptoms, but admitted them to be milder and with lower frequency. At the end of treatment, no patient reported severe or extremely severe symptoms. CONCLUSIONS: The results of this routine clinical practice program are consistent with data reported in controlled trials indicating the efficacy of esomeprazole even in patients with upper gastrointestinal symptoms "refractory" to treatments prescribed by primary care practitioners. PMID- 15800687 TI - Colonoscopy in Romania. AB - BACKGROUND: To date, colonoscopy is considered the gold standard for the investigation of the colon and also the gold standard method for colorectal cancer screening. AIM of this paper was to assess if and how is Romania prepared to cope with screening of colon cancer by means of colonoscopy. METHOD: We sent a study-type questionnaire addressed to all the centers in Romania known to perform digestive endoscopy and we inquired about the total number of colonoscopies and flexible sigmoidoscopies performed in 2003 (the questionnaire was sent to 43 centers). RESULTS: Thirty-eight centers responded to the question-naire. The total number of colonoscopies performed in Romania in 2003, obtained by collecting the data from the study centers, was 22,324. The number of sigmoidoscopies performed during the same period was 12,349. The ratio between the number of colonoscopies and sigmoidoscopies was 1.8/1. There were 106.3 colonoscopies /100,000 inhabitants. CONCLUSION: Considering the population of Romania (about 21 million inhabitants), the number of colonoscopies performed is insufficient for our country. The number of centers performing colonoscopy in Romania is also insufficient. PMID- 15800688 TI - Gastroparesis in diabetes mellitus: an ultrasonographic study. AB - BACKGROUND AND AIM: To investigate diabetic gastro-paresis, a complication of diabetes mellitus and its determinants. MATERIAL AND METHODS: A group of 36 patients with diabetes and 20 healthy controls, with comparable age and sex ratio were investigated with an established ultrasonographic method. A mixed test meal of 400 kcal was given. Antral fasting and postprandial area, postprandial distension and emptying time were assessed. Glucose control was estimated by the measurement of fasting and postprandial glucose as well as by assessing HbA1c. RESULTS: Antral area was nonsignificantly larger in diabetes mellitus. Postprandial antral area and postprandial antral distension were higher in diabetes than in the controls. The prevalence of gastroparesis was 52.7%, without differences between the two types of diabetes: 53.8% in type I and 52.2% in type II diabetes mellitus. Gastroparesis was associated with poor short and long time control of blood glucose and with positive parasympathetic tests. Gastroparesis did not correlate with symptoms. CONCLUSION: Gastroparesis is a real complication in diabetes mellitus and can be documented using an ultrasonographic method. Poor glucose control and autonomic neuropathy are associated with gastroparesis. PMID- 15800689 TI - Significance of anti-sacharomices cerevisiae antibodies (ASCA) in patients with inflammatory bowel diseases in Romania. AB - INTRODUCTION: Anti-Sacharomyces cerevisiae antibodies (ASCA) represent the immunologic marker correlated with Crohn's disease because of their high specificity (80-95%). The aim of this study is to confirm their value in a Romanian population with IBD. MATERIAL AND METHODS: A prospective longitudinal study was performed, which included patients admitted to Center of Gastroenterology and Hepatology Fundeni, Bucharest, in 2000 with ulcerative colitis (33 patients) - UC group, or Crohn's disease (40 patients) - CD group, and a control group (C) consisting of 22 healthy subjects. ASCA determination from serum samples was performed in the Erasmus University by ELISA technique. RESULTS: ASCA+ prevalence in CD group was 5 in 40 patients (12.5%), in UC group 0/33 (0%), 1/21 C group (4.9%), p=0.05. ASCA+ phenotype was found only in patients with CD diagnosed before the age of 40 years (A1), p=0.04. Also, ASCA+ phenotype correlated significantly with the colonic (L2) or ileocolonic (L3) extension (p=0.05). ASCA+ status did not correlate with the evolutive pattern of CD (stricturing, penetrating or non- stricturing non-penetrating), and neither did the clinical severity. CONCLUSIONS: ASCA+ prevalence in CD patients is much lower compared with North-American or West-European studies (12.5% versus 40 70%). In Romanian patients, ASCA assessment may be helpful in achieving a diagnosis of Crohn's disease, especially in those younger than 40 years, or in those with colonic or ileocolonic extension. PMID- 15800690 TI - Thymidine phosphorylase expression in patients with chronic viral hepatitis. AB - BACKGROUND: The role of thymidine phosphorylase (TP), an angiogenic factor, in chronic viral hepatitis is unclear. In the present study we investigated TP expression in chronic viral hepatitis due to hepatitis B virus (HBV) and hepatitis C virus (HCV) infection. METHODOLOGY: Fifty-eight patients suffering from chronic viral hepatitis and 13 cirrhotic patients due to either HBV or HCV were studied. TP expression was investigated using a standard immunohistochemical technique and the specific antibody P-GF.44C. RESULTS: The pattern of TP staining was predominantly nuclear, or mixed nuclear/cytoplasmic, but no significant difference in such expression was noted between chronic hepatitis B and C: nuclear immunostaining, t=0.0616, p=0.9511; cytoplasmic immunostaining, t=0.5188, p=0.6060. There was, however, a marked difference in TP expression between patients with chronic hepatitis B/C and cirrhosis. Notably, this finding applied to both nuclear (p=0.00754, p=0.0143, respectively) and cytoplasmic (p=0.000091, p=0.000031, respectively) expression. CONCLUSION: These observations indicate that TP expression, although linked to liver cirrhosis, may not be virus-related. PMID- 15800691 TI - Diagnosis and treatment of extrahepatic cholangiocarcinoma: results in a series of 124 patients. AB - AIM: The aim of the study was to evaluate the diagnostic methods and therapeutical results in patients with extrahepatic cholangiocarcinoma admitted between 2000 and 2003 and to assess the efficacy of various imaging methods. MATERIAL AND METHOD: We included 124 consecutive patients with suspected cholangiocarcinoma who underwent endoscopic retrograde cholangiopancreatography (ERCP) between 2000 and 2003. Every patient underwent an ultrasound (US) examination before ERCP. ERCP was considered as the "gold standard" for diagnosis. The therapy applied consisted of endoscopic stenting, US guided biliary drainage or surgery. The sensitivity, specificity and accuracy were calculated for every tumour localisation and also globally for all tumours. RESULTS: The sensitivity, specificity and accuracy of US were 85.9 %, 76.9 %, and 84.4 % for hilar localization, 59.1 %, 50 % and 57.1 % for the mid bile duct (CBD) and 33.3 %, 42.8 % and 36.8 % for the distal CBD tumours. The global performances for US in diagnosing extrahepatic cholangiocarcinoma were 73.5 %, 61.5 % and 70.9 %. In 73 cases (74.5%) an endoscopic stent was placed. In 11 cases (11.2%) we performed an US guided biliary drainage and in 14 cases (14.2%) surgery was recommended. CONCLUSIONS: Ultrasonography proved to be a reliable method for the diagnosis of cholangiocarcinoma in spite of its low accuracy for distal localization. The combination of the two investigations (US and ERCP) was very efficient in the management of these patients. Endoscopic stenting for hilar localization is a good therapeutical option. PMID- 15800692 TI - Ambulatory hemorrhoid therapy with radiofrequency coagulation. Clinical practice paper. AB - BACKGROUND: Despite availability of numerous surgical and non-surgical options for the treatment of hemorrhoids like sclerotherapy, rubber band ligation, cryosurgery, infrared photocoagulation, bipolar diathermy, and electro coagulation, none of these therapies has been acclaimed as the ultimate. Coagulation of hemorrhoids using a radio-frequency device is a new therapy to be added to the list. PATIENTS AND METHODS: In the present retrospective study, the early and long -term effects of radiofrequency coagulation on patients presenting with hemorrhoids is described. An Ellman radiofrequency generator was used for this procedure. In a separate, randomized, and blinded study, a comparative evaluation was carried out between radiofrequency coagulation and rubber band ligation in terms of their effectiveness and patient comfort. RESULTS: Two hundred and forty patients with Grade I and II hemorrhoids were treated by radiofrequency coagulation technique and were followed up for a period of 16 months. While 33 patients reported persistence or recurrence of bleeding, only few complained of pain or discomfort. The comparative study showed that though rubber band ligation is an effective procedure, its pain quotient is greater than the radiofrequency coagulation. CONCLUSION: This study shows that radiofrequency coagulation is an easy and effective alternative to conventional techniques employed in the treatment of bleeding hemorrhoids. It is easy to perform, is less painful, and has a low rate of complications. However, further results based on a longer follow-up of larger number of patients and its comparison with other conventional treatment techniques are called for. PMID- 15800693 TI - The emerging problem of nonalcoholic steatohepatitis (NASH). AB - Nonalcoholic steatohepatitis (NASH) is an increasing recognized form of chronic liver condition affecting both children and adults within the wide spectrum of fatty liver diseases. Recently NASH has been often associated with insulin resistance and has shown potential harmful evolution towards end-stage liver disease. Its incidence and prevalence is increasing, paralleling the rise in obesity and diabetes mellitus in Western Countries. Once all the other causes of persistent elevation of serum transaminase levels are excluded, the diagnosis of NASH can be only confirmed by liver histology. Non-invasive diagnostic tools, however, are awaited to allow the follow-up of patients at higher risk for major liver dysfunction. This article focus on current thoughts on the natural history and clinical presentation of NASH and describes current trends in the diagnosis and treatment of this emerging condition. PMID- 15800694 TI - Genetic factors in pancreatitis. AB - The understanding of pathogenesis of acute and chronic pancreatitis has benefited from the progress made in genetic investigations. The discoveries of the gain of function mutations of cationic trypsinogen gene (PRSS1) and the loss of function mutations of pancreatic secretory trypsin inhibitor (SPINK 1) or other potential defects in genes that regulate pancreatic secretory function or modulate inflammatory response to pancreatic injury has changed our current concepts on the pathogenesis of pancreatitis. Genetic factors play an important role in the susceptibility to pancreatic injury, severity and evolution of inflammatory process, leading in some cases to chronic inflammation and/or fibrosis. Acute pancreatitis is viewed as an event and chronic pancreatitis as a process, sequentially linked, reflecting a complex interaction between genetic and environmental factors. PMID- 15800695 TI - McKittrick-Wheelock syndrome - a rare cause of acute renal failure. AB - AIM: Fluid and electrolyte hypersecretion in the villous adenoma of the rectum is presented in the case of a 74 year old man presenting with a severe fluid imbalance. CASE REPORT: The patient had a 2-year history of mucous diarrhea and, on admission, presented prerenal uremia, hyponatremia and severe hypokalemia. At sigmoidoscopy, a 6/4 cm villous adenoma of the rectum was found. The increased loss of volume, followed by exhaustion of the physiological compensation mechanisms, led to a life-threatening hypokalemia, as well as to acute renal failure. Conservative treatment was followed by a temporary improvement of the renal function. Alternative treatment was: endocavitary irradiation, endoscopic resection and radical tumor surgery. The surgical removal of the adenoma led to complete recovery of the symptoms. CONCLUSION: The McKittrick-Wheelock syndrome can be a problem of difficult diagnosis, both for the gastroenterologist and also for the nephrologist. The patient may develop severe complications, which require a sustained treatment. PMID- 15800696 TI - Primary intraabdominal synovial sarcoma: a case report. AB - The authors report a case of intra-abdominal synovial sarcoma in a 41-year-old female. The tumor, which had an unusual location, the ascending mesocolon, had a mono-phasic spindle cell pattern. Immunohistochemical positivity for the epithelial membrane antigen (EMA) and cytokeratin AE1/AE3 differentiated the lesion from other spindle cell sarcomas with similar histology. The pathological and clinical features of the entity are briefly discussed. PMID- 15800697 TI - Hepatic tuberculomas. A case report. AB - BACKGROUND: The liver is often involved secondarily by sytemic/extrahepatic vascular, metabolic, infectious, granulomatous, and malignant processes. CASE REPORT: A 57-year old woman was admitted because of fatigue, malaise, night sweating and right upper abdominal quadrant pain. Physical findings revealed pale skin and mucosae, and hepatomegaly. Laboratory data showed normocytic, normochromic anemia and marked inflammatory syndrome. Real-time ultrasonography revealed one hypo-echogenic nodule, 25 mm in size, in the segment II of the liver, and three hypoechogenic nodules, 11-25 mm in diameter, in segments III and IV. Enlarged lymph nodes, 20-50 mm, were observed in the region of the pancreatic head. Histo-pathological examination of the liver and peritoneum biopsies evidenced granulomas with caseous necrosis and multinuclear Langhans' giant cells, indicating hepatic and peritoneal tuberculosis. The tuberculostatic chemotherapy (isoniazid 300 mg/24h; rifampycin 600 mg/24h and pyrazinamid 2000 mg/24h) was applied for 12 months. The patient responded well to the treatment, followed-up for 6-12 months. CONCLUSION: In the differential diagnosis of focal liver lesions, pseudotumoral hepatic tuberculomas must be considered. PMID- 15800698 TI - Complicated endoscopic management of Bouveret's syndrome. A case report and review. AB - A rare case of gastric outlet obstruction due to a large gallstone (Bouveret's syndrome) presenting with abdominal pain and vomiting is reported. The endoscopic management of the gallstone was unsuccessful because of the size of the stone that got stuck in oesophagus. The final management of the patient was surgical and the gallstone was retrieved via a gastrotomy. The case report is followed by a brief review of Bouveret's syndrome. PMID- 15800699 TI - Endoscopic clipping focused on "triclip" for bleeding Dieulafoy's lesion in the colon. AB - Endoclips were among the earliest accessories developed for gastrointestinal endotherapy. Now they are currently used for the control of gastrointestinal bleeding and for the closure of perforations, fistulas, and anastomotic leaks. Numerous technical improvements of the original device have led to the development of new devices: rotatable clip, preloaded hemoclip, TriClip and repositioning clip device. The endoclip devices are easy to use. The technique comprises three steps: exposing the clip from the sheath, opening the clip to the maximum, and deploying the clip. Dieulafoy's lesion is an abnormal, submucosal "caliber-persistent artery" that protrudes through a minute mucosal defect. Although it accounts account for up to 5% of acute upper gastrointestinal bleeding, only a few cases involving colo-rectum have been reported. We report a case of Dieulafoy's lesion of the colon which has given us the opportunity to briefly review the indications and technique of endoclipping. PMID- 15800700 TI - Ultrasonography of acute pancreatitis -- an essay in images. AB - Acute pancreatitis is a relatively frequent disorder with an increased risk potential. Ultrasound may represent the first imaging technique to be used in disease evaluation. The diagnosis is usually made based on the increase in pancreas volume and on the changes in structure detected on the ultrasound examination. The most frequent complications are: local and distal collections, parenchyma necrosis, superinfection and vascular involvement. PMID- 15800701 TI - Liver biopsy under ultrasound control for diffuse liver disease -- toward a faster, safer, cost-effective and easy acceptable procedure. PMID- 15800702 TI - Subclinical thyroid disorders and inflammatory bowel disease. PMID- 15800703 TI - Progression of liver fibrosis in young blood donors infected with hepatitis C virus: different predictive factors? PMID- 15800705 TI - Clinical results of intensity-modulated radiation therapy (IMRT) for tumors of the head and neck region. AB - Intensity-modulated radiation therapy (IMRT) is an advanced form of the three dimensional conformal radiation therapy (3D-CRT). Highly conformal dose distribution is the basic feature of IMRT. The head and neck region is suitable for this new technology since the primary tumor is often surrounded by several critical structures. IMRT offers the ability of dose escalation due to steep dose gradient towards healthy tissues. In this review, clinical results of IMRT in several head and neck sites are presented, including intracranial tumors. Parotid sparing strategies and patterns of local-regional failures are analyzed. The possibilities of irradiation of recurrent malignancies are mentioned. In perspective, the potential of IMRT should be explored in conjunction with altered fractionation regimens, including simultaneous integrated boost (SIB). Particularly, studies with dose escalation are desirable. PMID- 15800706 TI - The prognostic role of clinical, morphological and molecular markers in oral squamous cell tumors. AB - Despite of considerable advances in the diagnostic and therapeutic possibilities, the prognosis of epithelial tumors in the oral cavity is still very poor. A knowledge of the prognostic factors at the beginning of treatment is therefore indispensable for determination of the appropriate therapy for the given patient. These factors may be linked to the patient (e.g. age, sex, general condition and immunological parameters) or to the tumor localization. A survey of the literature reveals that the TNM stage, the grade, the mode of invasion and the depth of the tumor infiltration are generally the most important factors influencing the fate of the patient. The prognosis depends primarily on the clinicopathological parameters, though even if they are known, it is not possible to screen out those patients who are at particular risk of a relapse. During the past 10 years, study of the DNA content distribution, the proliferation markers and certain oncogenes has come into the focus of attention; great interest is also shown in the extracellular matrix components and the metalloproteinases, which play key roles in the invasion and metastasis formation. PMID- 15800707 TI - Peritoneal cytology in endometrial cancer. AB - The results as well as prognostic interpretation of peritoneal washing cytology in endometrial cancer cases is still controversial. The incidence rate of positive cytology varies widely and the clinical significance of the positive results, especially in patients with early stage of endometrial cancer remains also controversial. Prognostic significance of malignant peritoneal cytology in endometrial cancer patients in comparison with those with ovarian cancer has been less well defined. At present, positive peritoneal cytology is not a negative prognostic factor itself, but it enhance other negative prognostic indicators. Literature regarding the significance of peritoneal cytology in endometrial cancer was reviewed in order to draw conclusions for possible therapeutic implications in patients with positive cytologic findings. PMID- 15800708 TI - Bcl-2 expression in acute myelogenous leukemia: the relation to myeloid antigen expression and response to therapy in Iranian patients. AB - Product of Bcl-2 gene prolongs survival of hematopoietic cells by inhibition of programmed cell death. The aim of this study is to examine the expression of the bcl-2 protein in a group of patients with AML and its relation to clinical features and response to therapy. Slides from the bone marrow or peripheral blood of 70 patients with AML were assessed for the expression of bcl-2 by immunocytochemistry. The expression of myeloid and non-lineage associated markers was detected by indirect immunofluorescence method. Correlation between bcl-2 and markers expression and patients characteristics was determined. More than 20% positivity for bcl-2 was found in 22 (31.4%) patients. Bcl-2 expression showed an association with M4 and M5 subtypes (p<0.01) and was correlated with clinical parameters including WBC and platelet count, extramedullary disease and Hb level. Bcl-2 expressing cells were significantly higher in CD15(+) and CD13(+) patients and lower in CD11b(+) and CD33(+) cases (p<0.001). Complete remission (CR) rate was significantly lower in cases with 20% or more bcl-2 positivity than others (24.4% v 75.6%). A shorter CR duration was observed in bcl-2+ patients when compared with bcl-2- ones (571+/-50 versus 850+/-17 days)(p=0.0001). The expression of bcl-2 was also associated with shorter survival (p=0.0001). Survival time for bcl-2+ patients was 831+/-44 days versus 1119+/-17 days for bcl 2- ones. CD11b and CD33 positivity was associated with longer survival whereas CD13 and CD15 positivity was correlated with lower survival (p<0.007). In multivariate analysis bcl-2 positivity was associated with poor survival. Bcl-2 expression showed a prognostic value in our patients indicating that even despite of some differences in treatment regimen, immunocytochemical analysis of this marker is still a simple and inexpensive method for evaluation of prognosis in AML patients. Bcl-2 expression may be related to the expression of differentiation associated markers. PMID- 15800709 TI - CYP19 gene expression and aromatase activity in endometrial cancer tissue: importance of the type of the disease. AB - Aromatase (CYP19) activity in malignant endometrium presents local mechanism with potential ability to support tumor growth. The data on interrelation between activity of this enzyme and its mRNA signal in endometrial cancer (EC) tissue are very scarce and inconclusive. To correct this gap we studied aromatase activity and gene expression totally in 19 samples of EC (17 of them -- from postmenopausal women) collected during surgery. Aromatase activity was evaluated by tritium water release assay and CYP19 gene expression -- with rt-PCR. Additionally, all studied EC cases were divided on the basis of case history and some characteristics of host and tumor and in accordance with existing classification into cases belonging to type I or II of the disease (correspondingly, 13 and 6 observations). Positive correlation between enzyme activity and CYP19 mRNA signal (R(s)=+0.63, p<0.05) was revealed in the whole group of studied samples. Aromatase activity varied in evaluated material between 1.45 fM/mg prot./hr and 11.49 fM/mg prot./hr, and in type II cases it was higher (7.27+/-0.96 fM/mg prot/hr) than in type I observations (4.96+/-0.66 fM/mg prot./hr, p=0.066). CYP19 mRNA expression was not revealed in 6 cases and all of them belonged to the type I of disease. Thus, although type II of EC is frequently considered as hormone non-dependent, increased ability of this tumor type to estrogen biosynthesis (on CYP19 gene and protein level) may lead to reconsideration of such conclusion and warrants further investigation. PMID- 15800710 TI - Disease status in patients with chronic myeloid leukemia is better characterized by BCR-ABL/BCR transcript ratio than by BCR-ABL transcript level, which may suggest a role of normal BCR gene in the disease pathogenesis. AB - Monitoring of BCR-ABL transcript level is widely used in chronic myeloid leukemia (CML) to follow up response to therapy. In this study we compare abilities of two quantitative RT-PCR assays to characterize the disease status in CML patients: RT PCR quantifying the BCR-ABL transcript concentration and RT-PCR determining the BCR-ABL/BCR transcript ratio (R). We demonstrate that in non-responders only R, but not BCR-ABL, unambiguously characterizes the state of disease. Moreover, R values >1 found in all poor responders indicate lower BCR expression compared to BCR- ABL in these patients. Our results demonstrate the importance of BCR-ABL/BCR transcript ratio for the disease status and the disease prognosis characterization and suggest a possible role of the normal BCR gene expression in CML pathogenesis. PMID- 15800712 TI - Significance of qualitative PCR detection method for preemptive therapy of cytomegalovirus infection in patients after allogeneic hematopoietic stem cell transplantation -- single-centre experience. AB - Both early cytomegalovirus (CMV) monitoring and prophylactic antiviral therapy can decrease clinical complications or can prevent them in patients after allogeneic hematopoietic stem cell transplantation (HSCT). Presented paper summarizes experiences with using regular monitoring of reactivation of CMV after allogeneic HSCT by qualitative polymerase chain reaction (PCR) method to prevent the development of symptomatic CMV disease. Samples of peripheral blood leukocytes (PBL) in 71 patients were monitored. Because of retransplantation in two patients, 73 transplantations, each followed by the monitoring, were performed. Patients were monitored weekly after the transplantation for CMV DNA emia in PBL. An episode of CMV infection representing an indication for preemptive ganciclovir (GCV) or foscarnet (FOS) therapy was defined as two consecutive positive PCR results in 4-7 days. Median time of monitoring was 313 days. The CMV infection was found in 28/73 monitorings (38.4%) and always was followed by preemptive therapy. One recurrence of CMV infection was observed in 4/28 (14.3%) monitorings and two recurrences in 1/28 (3.6%) monitorings. Presented approach resulted in complete prevention of overt CMV disease and this study enable to show that qualitative PCR method for determination of incipient CMV infection followed by preemptive therapy is suitable for preventing patients after allogeneic transplantation from CMV disease. PMID- 15800711 TI - Peroxisome proliferator-activated receptors (PPAR) agonists affect cell viability, apoptosis and expression of cell cycle related proteins in cell lines of glial brain tumors. AB - The nuclear receptors PPARs (peroxisome proliferator-activated receptors) are transcription factors activated by specific ligands. PPARs play an important role in carcinogenesis, inflammation, atherosclerosis, lipid metabolism and diabetes. There is evidence that activation of PPARs by specific ligands is able to suppress the growth of different types of human cancer by mechanisms including the growth arrest, apoptosis and induction of differentiation, although the detailed signalling pathways have not been completely elucidated to date. The aim of our study was to determine whether synthetic ligands of PPARalpha and PPARgamma could affect the viability, proliferation, differentiation, apoptosis and expression of some cell cycle related proteins in glial tumor cell lines. The study was performed on human glioblastoma cell lines U-87 MG, T98G, A172 and U 118 MG. Cell lines were treated by ligands of PPARalpha (bezafibrate, gemfibrozil) and PPARgamma (ciglitazone). MTT, flow cytometry, TUNEL assay and immunoblotting were used for detection of changes in cell viability, proliferation, differentiation and apoptosis. Bezafibrate, ciglitazone and gemfibrozil inhibited viability of glioblastoma cell lines. The synthetic ligands significantly reduced or induced the expression of cyclins, p27Kip1, p21Waf1/Cip1, MDM-2, Bcl-2, Bax, PARP, Caspase 3, androgen receptors, etc. and did not affect the expression of the differentiation marker GFAP. Flow cytometry confirmed arrest of the cell cycle although the detection of apoptosis was controversial. Apart from hypolipidemic and hypoglycaemic effects, PPAR ligands may also have significant cytostatic effects of potential use in anticancer treatment. PMID- 15800713 TI - Split-course accelerated hyperfractionation (CHA-CHA) for advanced head and neck cancers -- preliminary results. AB - The purpose of the study was to evaluate toxicity and effectiveness of the split course, accelerated hyperfractionation with a planned gap (CHA-CHA) on a base of the pilot group treatment results. The material consisted of 27 patients with advanced (T2N2c-3, T3-4N2), oral cavity, oro- hypopharynx and larynx cancers, irradiated with a df 1.6 Gy twice a day, with 6-hour gap, up to TD 64 Gy in 28 days, with 8-day gap in a middle. Mean tumor dimensions were 4.2 and 3.9 cm in X and Y axes, respectively and mean nodes dimensions 3.9 cm in both axes. The course and intensity of the mucositis were evaluated in the Dische scale together with the degree of a tumor and nodes regression. Survival analysis and dependencies evaluation between physical/biological factors and treatment results were performed. The mucositis intensity was high from 2nd to 5th week (the highest mean - 15 Dische points in 4th week), but on acceptable level. 54.5% of CR of the tumor and 57.1% of nodes were noted. Survival analysis showed 44% of a 2-year and 35% of a 4-year OS and 44% of a 2-year and 38% of a 4-year survival excluding deaths from distant metastases. Correlation between the hemoglobin level and the nodal regression two months after the radiotherapy (p=0.01), showed the tumor diameter and TCP (p=0.047), the OS and the nodes diameter (p=0.004), the PS (p=0.02) and the hemoglobin level (p=0.04). CHA-CHA could be a valid method in advanced head and neck cancer treatment but its efficacy should be confirmed in randomized phase of trial. PMID- 15800714 TI - Cerebral metastases of malignant melanoma: contemporary treatment modalities and survival outcome. AB - The aim of our study was to analyze prognostic factors, effect of treatment and survival outcome of a contemporary cohort of melanoma patients with cerebral metastases and eventually propose new recommendations regarding therapy. Sixty four patients with melanoma brain metastases were treated in our department within a 15-year period. We performed a retrospective analysis of their survival with respect to the type of treatment instituted. Four groups were formed according to treatment: Group A patients treated with surgery followed by radiotherapy; group B temozolomide as first-line treatment and radiotherapy after cerebral disease progression; group C radiotherapy alone; group D supportive care only. Patients* characteristics influenced the selection of treatment modality: Group A (7.8%) patients with a single brain metastasis (p=0.001) and controlled extra-cranial disease (p<0.0001), while Group D (21.8%) patients with ulcerated primary lesions (p=0.010) and uncontrolled extra-cranial disease (p<0.0001). Only group B (26.6%) and C (43.7%) patients with similar characteristics including more than one brain lesion. Median overall survival was 3 months. In univariate analysis, median survival for groups A, B, C and D was 12, 5, 3 and 2 months, respectively (p<0.0001). The survival difference between the surgery and non surgery groups was statistically significant (p=0.0011). Patients treated with supportive care had the worse prognosis (p<0.0001). A survival benefit for patients receiving first-line treatment with temozolomide, as compared to those receiving radiotherapy alone was noted (p=0.0267). In multivariate survival analysis, the number of brain lesions (p=0.0138), the absence of uncontrolled extra-cranial disease (p=0.00221) and the type of treatment for the cerebral disease (p=0.0053) remained significant independent survival predictors. Patients' characteristics remain a critical factor for treatment selection. The number of brain metastases, the extent of disease and the type of treatment represent independent survival predictors. Melanoma patients with a single brain metastasis and controlled extra-cranial disease gain a survival benefit, if surgically treated. Including temozolomide in the first-line treatment of melanoma patients with brain metastases who would have been treated with radiotherapy alone, might present a promising future direction affecting the length of survival. PMID- 15800715 TI - Prevention of febrile neutropenia in cancer patients by probiotic strain Enterococcus faecium M-74. Pilot study phase I. AB - Febrile neutropenia (FN) remains a potentially life-threatening complication of anticancer chemotherapy. Bacterial translocation via intestinal mucosa is a significant mechanism of FN development. Competitive inhibition of bowel colonization by pathogenic microorganisms by lactic acid bacteria could be a useful prevention of FN. The aim of the study was the evaluation of dose and safety of probiotic strain Enterococcus faecium M-74 enriched with organic selenium in patients with solid and hematological malignancies. Eleven (9 M/2F) patients were included in the study. In the first phase six patients with germ cell tumors treated by chemotherapy were included. They received prophylaxis by nonpathogenic strain E. faecium M-74 during 2 cycles of chemotherapy. The planned daily dose was 6 x 10(9) bacteria. Regarding the insufficient colonization of the gut, the dose was further increased up to 18 x 10(9) tid. After safety evaluation, five patients were included with relapse of acute leukemia. In patients with germ cell cancer, severe neutropenia G3/4 was noted in 10 of 12 cycles of chemotherapy. The febrile episode was not observed in any of the patients. The gut colonization by enterococci reaches 10(6) CFU/g stool. In 5 patients with acute leukemia during 127 days of severe neutropenia 12 febrile episodes occurred. There was not noted any febrile episode or infection provoked by the tested strain. Tolerance of therapy was excellent without significant undesirable effects. Optimal dose was assessed and safety of probiotic strain was evaluated in neutropenic patients with solid, or hematological malignancies. Based on these results we plan phase II study to evaluate the effectiveness of this strain in FN prophylaxis. PMID- 15800716 TI - Inoperable oropharyngeal carcinoma treated with concomitant irradiation, mitomycin C and bleomycin - long term results. AB - Patients with inoperable head and neck tumors were treated concomitantly with radiochemotherapy with mitomycin C and bleomycin in our prospective randomized clinical trial (1991- 1993). For the subgroup of patients with oropharyngeal carcinoma the results with radiochemotherapy were significantly superior to irradiation alone. Such scheme of treatment was then adopted as standard method. Here we present the long-term results and dose- response relationships in patients with inoperable oropharyngeal carcinoma treated by the same radiochemotherapy scheme till 1997. Ninety-five patients with stage III and IV inoperable oropharyngeal squamous cell carcinoma were treated with curative intent, concomitantly with supra-voltage irradiation 2 Gy/day 5 times weekly to 60-73 Gy, bleomycin 5 mg 2 times weekly and. one application of mitomycin C 15 mg/m(2) after 10 Gy. Logistic dose- response curve was calculated. Median follow up was 85 months. The loco-regional control, disease- free survival and overall survival at 5 years were 55%, 51% and 32% (95% CI: 44-67%, 41-62%, 22-42%), respectively. The probability of new primary malignancy at 5 years was 23%. In multivariate analysis performance status, biological equivalent dose, dose of bleomycin, and stage were identified as independent prognostic factors for loco regional control, disease-free, and overall survival. Th gamma-value of dose response curve was 2.86. The outcome of the disease was directly proportional to intensity of irradiation and chemotherapy. It appears that in our concomitant radiochemotherapy MiC increased radioresponsiveness of the tumor by its effect on hypoxic fraction. PMID- 15800717 TI - The influence of thalidomide therapy on cytokine secretion, immunophenotype, BCL 2 expression and microvessel density in patients with resistant or relapsed multiple myeloma. AB - Thalidomide (THAL) is currently used as a novel drug in patients with chemotherapy resistant or relapsed multiple myeloma. THAL antitumor activity seems to be very complex, however the precise mechanisms of its action are still not fully understood. The aim of this study was to assess some of possible mechanisms of THAL action both in in vivo analysis of immune cells phenotype and in in vitro cultures with THAL. The study involved 30 patients with relapsed or chemotherapy refractory multiple myeloma who were qualified to THAL treatment. We assessed immunophenotype of malignant plasma cells and T lymphocytes in both peripheral blood (PB) and bone marrow (BM) samples taken before and after 4 and 8 weeks of THAL treatment. Before therapy cytokine secretion (VEGF, HGF, bFGF, TNF, IL-6 an sIL-6R) and Bcl-2 expression in PB and BM cell cultures with THAL were analyzed. We used flow cytometry technique and ELISA method. The clinical response to therapy was assessed after 4 and 8 weeks of treatment. We also investigated microvessel density (MVD) in bone marrow samples before the THAL treatment and after 6 months of therapy in the group of responding patients. In cell cultures with THAL we detected statistically significant lowering of analyzed cytokines concentration and the decrease in Bcl-2 expression by malignant plasma cells in BM and CD8(+) T lymphocytes in BM and PB. In the group of patients responding to therapy we observed the decrease in the number of myeloma cells and significant increase of CD4(+) and CD8(+) cells in both PB and BM samples. There was statistically significant increase of CD3(+)/CD69(+) cells in the course of therapy, while the percentage of CD3+/HLA-DR(+) cells was significantly lower after 8 weeks of therapy. We also detected lowering of MVD after THAL therapy in responders group. The obtained results demonstrate that THAL efficacy in MM is multidirected and included such mechanisms like down regulation of proangiogenic cytokines, that could lead to lowering of MVD, induction of apoptosis and influence on malignant cells and T lymphocytes immunophenotype. PMID- 15800718 TI - Endostatin serum level in acute myeloid leukemia. AB - Increased levels of tumor angiogenesis have been demonstrated in variety of solid tumors and hematological malignancies including acute myeloid leukemia (AML). The aim of the study was to evaluate serum level of endostatin in newly diagnosed patients with AML before chemotherapy and after achieving complete remission (CR). Serum samples from 68 adult patients (28 females and 40 males, median age 42 years, range 21-83 years) with AML had been taken before chemotherapy was administered. In addition 21 out of 68 patient were analyzed again after achieving CR. Endostatin levels were measured using ChemiKine sandwich ELISA kit (Chemicon International). Twelve samples from healthy volunteers (5 females and 7 males, median age 40 years; range 35-65 years) were evaluated as the control. Endostatin serum levels were significantly higher in untreated AML patients than in the normal controls. In AML patients baseline endostatin levels were significantly lower than in CR. We did not found any correlation between white cell count or percentage of blasts in the bone marrow and endostatin level. Moreover endostatin levels did not differ statistically among AML FAB subgroups. Increased endostatin plasma levels may reflect intensity of inhibition of angiogenesis and may by useful in prognosis of CR in AML. Chemotherapy can modulate the regulation of angiogenesis in AML patients. PMID- 15800719 TI - SARS: two years on... PMID- 15800720 TI - Biostatistics 304. Cluster analysis. PMID- 15800721 TI - Presentation of severe acute respiratory syndrome (SARS) patients in a screening centre. AB - INTRODUCTION: On 22 March 2003, the Ministry of Health, Singapore, designated Tan Tock Seng Hospital as the nationwide severe acute respiratory syndrome (SARS) hospital and its Emergency Department (ED) took over the role as the screening center for SARS on 26 March 2003. We describe the initial clinical characteristics of probable or suspect SARS patients that presented to the ED. METHODS: A retrospective study of patients who were admitted through the ED and subsequently diagnosed to have probable SARS and suspect SARS was done. The data of these patients from the ED log were reviewed and analysed. RESULTS: From 13 March 2003 to 31 May 2003, 11,461 patients were screened for SARS and 1,386 patients were admitted. Of these, 117 patients were diagnosed to have probable SARS and 146 suspect SARS. Their mean age was 36.7 years (range 1-80). Among these patients, there were 122 men (46.4 percent), and 205 were Singaporeans (77.9 percent). 13 patients had no initial contact history upon presentation to the ED. The mean duration between onset of symptom to presentation to the ED was 3.1 days. Travel history was less common in probable SARS cases than in suspect SARS cases as the epidemic was due mainly to local transmission. Fever was the most common presenting symptom (91.6 percent), and gastrointestinal symptoms were the least (6.9 percent). In the ED, 249 (94.7 percent) patients had chest radiographs and 86 (32.7 percent) had full blood count done. 22.2 percent of probable SARS patients had normal chest radiographs when they first presented to the ED. CONCLUSION: The World Health Organisation criteria were important screening tools and admission guides, but should not be strictly followed. It was difficult to differentiate between probable and suspect SARS patients in the ED. PMID- 15800722 TI - Surveillance of severe acute respiratory syndrome (SARS) in the post-outbreak period. AB - INTRODUCTION: This retrospective one-month survey evaluated the practicality of post-severe acute respiratory syndrome (SARS) surveillance recommendations in previously SARS-affected countries, namely Singapore. These included staff medical sick leave for febrile illness, inpatient fevers, inpatient pneumonia, atypical pneumonia, febrile illnesses with significant travel history and sudden unexplained deaths from pneumonia/ adult respiratory distress syndrome (ARDS). METHODS: Surveillance data on medical sick leave of staff, all inpatient fevers, all febrile (temperature greater than or equal to 38 degrees Celsius) inpatient pneumonia, including atypical pneumonia, and deaths from pneumonia were collected from sick leave reports, ward reports, isolation room rounds and mortuary reports from 1 to 28 September 2003. RESULTS: Baseline results show 167 (1.4/1000 staff days) observed in staff sick leave for febrile illnesses, and 1798 (71.3/1000 bed days) observed for inpatient fever. There were 40, 31 and 12 instances, respectively, of staff having temperatures of high fever (greater than or equal to 38 degrees Celsius), prolonged sick leave (3 days or more), and repeated sick leave (within 7 days) for febrile illnesses. An average of 4.6 wards a day potentially fulfilled the World Health Organisation SARS alert criteria. Of 27 cases with fever, pneumonia and a total white count of less than 10,000 cells per cubic mm as per Ministry of Health, Singapore criteria for the diagnosis of atypical pneumonia, only five were identified by clinicians. CONCLUSION: Surveillance is time-consuming and current recommendations are not specific enough to be used practically. Surveillance indicators for inpatients must overcome a high degree of background noise. PMID- 15800723 TI - Otorhinolaryngeal foreign bodies in children presenting to the emergency department. AB - INTRODUCTION: Accidents with foreign bodies are common in the paediatric population. It is impossible to mandate that all foreign bodies (FB) in the ear, nose and throat (ENT) of children should be removed by the specialty-trained physicians. This study evaluates the management of ENT FB removal in children achieved by emergency physicians not trained in otolaryngology in an urban tertiary care paediatric emergency department. METHODS: A retrospective study was conducted on consecutive paediatric patients presenting with suspected foreign body in the ear, nose or throat to the children's emergency department (ED) of KK Women's and Children's Hospital over a 10-month period. Removal methods, foreign body types, rates of successful removal and associated complications were evaluated. RESULTS: There were 353 patients, most of whom presented after office hours. An attempt at removal of FB by the emergency physician was made in 76.8 percent of the cases. ENT specialist referral in the ED was made in 1.7 percent of the cases. 50.1 percent of cases were discharged after successful removal of FB in the ED. 4.2 percent of cases were admitted for removal of FB and 44.8 percent of cases were referred to the ENT specialist clinic for further assessment. CONCLUSION: The emergency physician managed most cases in the ED and urgent referral to ENT specialists was not required. Complications and morbidity often occur from repeated attempts at removal of the FB. ENT opinion should be sought whenever there is doubt. The ED physician should be skilled in techniques of FB removal, especially throat FB, which had the lowest rate of success in our study. PMID- 15800724 TI - Kimura's disease: a diagnostic and therapeutic challenge. AB - INTRODUCTION: Kimura's disease (KD) is a rare, benign, chronic inflammatory disease with unknown aetiology. Its manifestation is protean. KD has a predilection for the head and neck area, and typically presents as tumour-like lesions that could be easily misdiagnosed. We review our experience with four recent cases. METHODS: Over a four-year period, all patients admitted to Singapore General Hospital with KD of the head and neck region were retrospectively reviewed. Biodata, presenting symptoms and clinical parameters, especially serum eosinophil levels, preoperative investigations, type of surgical procedures and outcome were documented. RESULTS: Four patients presented with KD of the head and neck and displayed varied manifestations of the disease. All the patients had raised serum eosinophil levels. None of them had renal involvement. Preoperative computed tomography were performed in two of the patients and showed features suggestive of KD. Fine-needle aspiration cytology that was performed in two patients was not useful in the diagnosis. All the patients underwent surgical excision of the lesions. Only one patient had multiple recurrence, both at the original and remote sites in the head and neck. CONCLUSION: The clinical presentation and behaviour of KD is very variable. Preoperative imaging is useful in the diagnosis of the disease but the final diagnosis is histological. Surgical excision is the current treatment of choice but recurrence is common. A high index of suspicion and awareness is vital in the early diagnosis and management of KD. PMID- 15800725 TI - Circadian rhythmicity of plasma lipid peroxidation and antioxidants in oral squamous cell carcinoma. AB - INTRODUCTION: Oral cancer is one of the leading cancers in India accounting for 30 to 40 percent of all cancers. Disturbances in lipid peroxidation and antioxidants status have been implicated in the pathogenesis of several cancers including oral cancer. However, circadian disturbances of oxidants and antioxidants in oral cancer patients were not reported. METHODS: The levels of plasma thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) and activity of glutathione peroxidase (GPx) in ten oral cancer patients and an equal number of age-matched healthy subjects were assayed at every 6 hour intervals using colorimetric methods and their circadian characteristics were analysed using Cosinorwin computer software programme. RESULTS: Alterations in mesor, amplitude, acrophase and r value of the chosen parameters were noticed. DISCUSSION: The desynchronisation of plasma thiobarbituric acid reactive substances and the altered circadian characteristics of antioxidants observed in this study, may deserve further investigation for the early diagnosis, prognosis and for the efficacy of cancer chronotherapy. PMID- 15800726 TI - Missed posterior fracture-dislocation of the humeral head following an electrocution injury to the arm. AB - Posterior dislocation of the shoulder is a rare and commonly missed injury. While prompt diagnosis and treatment is important to prevent untoward sequelae, it is often diagnosed and treated too late. The first reported case of a posterior fracture-dislocation of the humeral head following a domestic electrocution accident in Singapore is described in a 52-year-old man. The injury was missed by several doctors before a humeral head replacement was done. The diagnostic pitfalls and management of this injury and ways to avoid a missed or delayed diagnosis are discussed. PMID- 15800727 TI - Multifocal pigmented villonodular synovitis in a child. AB - Pigmented villonodular synovitis is a rare benign disorder and usually affects young and middle-aged adults. It occurs either as a localised pedunculated form or more common diffuse form, and is almost always unifocal. Only few cases of multifocal involvement have been reported, all of them in children. Multifocal pigmented villonodular synovitis occurring in a 5-year-old girl is presented. Both her knees and left elbow were involved. All three lesions were of localised pedunculated type and were completely excised. She is believed to be one of the youngest patients reported. PMID- 15800728 TI - Painless scrotal swelling: ultrasonographical features with pathological correlation. AB - Scrotal swelling may be due to extratesticular and intratesticular lesions. The majority of extratesticular lesions are benign while the majority of intratesticular lesions are malignant. Ultrasonography (US) is helpful in separating extra- from intratesticular lesions. US can show whether a mass is cystic, solid or complex, and also features such as associated calcifications, epididymal involvement, scrotal skin thickening and colour Doppler flow pattern. Extratesticular lesions include hydrocoele, spermatocoele, varicocoele, epididymal cyst, hernia and tumours of the epididymis and cord structures. Intratesticular lesions include primary tumour, metastases, lymphoma and leukaemia. Tuberculous epididymitis or epididymo-orchitis may also present with painless scrotal swelling. US features of these disease patterns, with pathological correlation, are presented in this pictorial essay. PMID- 15800730 TI - Selection pressure-driven aerobic granulation in a sequencing batch reactor. AB - In recent years, the research on aerobic granulation has been intensive. So far, almost all aerobic granules can form only in sequencing batch reactors (SBR), while the reason is not yet understood. This paper attempts to review the factors involved in aerobic granulation in SBR, including substrate composition, organic loading rate, hydrodynamic shear force, feast-famine regime, feeding strategy, dissolved oxygen, reactor configuration, solids retention time, cycle time, settling time and exchange ratio. The major selection pressures responsible for aerobic granulation are identified as the settling time and exchange ratio. A concept of the minimal settling velocity of bioparticles is proposed; and it is quantitatively demonstrated that the effects of settling time and exchange ratio on aerobic granulation in SBR can be interpreted and unified on the basis of this concept very well. It appears that the formation and characteristics of aerobic granules can be manipulated through properly adjusting either the settling time or the exchange ratio in SBR. Consequently, theoretical and experimental evidence point to the fact that aerobic granulation is a selection pressure-driven cell-to cell immobilization process. PMID- 15800731 TI - Different control mechanisms regulate glucoamylase and protease gene transcription in Aspergillus oryzae in solid-state and submerged fermentation. AB - Solid-state fermentation (SSF) with Aspergillus oryzae results in high levels of secreted protein. However, control mechanisms of gene expression in SSF have been only poorly studied. In this study we show that both glucoamylase (glaB) and protease (alpA, nptB) genes are highly expressed during surface cultivation on wheat-based solid medium, and even higher during cultivation on wheat kernels. In wheat-based liquid medium, low levels of gene expression are observed. Typical SSF cultivation conditions, such as low water activity and the formation of aerial hyphae, did not contribute to the high-level gene expression on wheat based solid medium. Analysis of wheat-based solid and liquid cultivations showed differences in carbon and nitrogen utilisation and external pH. The results presented show that the difference in regulation of transcription of the alpA and nptB genes in wheat-based liquid and solid medium could be pH dependent, involving a pH-dependent transcription regulator. The results obtained suggest that the difference in regulation of transcription of the glaB gene in wheat based liquid and solid medium is caused by a difference in carbohydrate degradation and consumption under the different culture conditions. PMID- 15800732 TI - Production of 3-hydroxy-n-phenylalkanoic acids by a genetically engineered strain of Pseudomonas putida. AB - Overexpression of the gene encoding the poly-3-hydroxy-n-phenylalkanoate (PHPhA) depolymerase (phaZ) in Pseudomonas putida U avoids the accumulation of these polymers as storage granules. In this recombinant strain, the 3-OH-acyl-CoA derivatives released from the different aliphatic or aromatic poly-3 hydroxyalkanoates (PHAs) are catabolized through the beta-oxidation pathway and transformed into general metabolites (acetyl-CoA, succinyl-CoA, phenylacetyl-CoA) or into non-metabolizable end-products (cinnamoyl-CoA). Taking into account the biochemical, pharmaceutical and industrial interest of some PHA catabolites (i.e., 3-OH-PhAs), we designed a genetically engineered strain of P. putida U (P. putida U DeltafadBA-phaZ) that efficiently bioconverts (more than 80%) different n-phenylalkanoic acids into their 3-hydroxyderivatives and excretes these compounds into the culture broth. PMID- 15800733 TI - Anatomical variations of the second thoracic ganglion. AB - In recent years the second thoracic ganglion has gained anatomical significance as an important conduit for sympathetic innervation of the upper extremity. Thoracoscopic excision of the second thoracic ganglion is now widely recognized as affording the most effective treatment option for palmar hyperhidrosis. This study recorded the incidence, location and associated additional neural connections of the second thoracic ganglion. Bilateral dissection of 20 adult cadavers was undertaken, and all neural connections of the second thoracic ganglion were recorded. Nineteen cadavers (95%) demonstrated additional neural connections between the first thoracic ventral ramus and second intercostal nerve. These were classified as either type A (47.5%) or type B (45%) using the intrathoracic ramus (nerve of Kuntz) between the second intercostal nerve and the ventral ramus of the first thoracic nerve as a basis on both right and left sides. The second thoracic ganglion was commonly located (92.5%) in the second intercostal space at the level of the intervertebral disc between the second and third thoracic vertebrae. Fused ganglia between the second thoracic and first thoracic (5%) and stellate (5%) ganglia were noted. These findings should assist the operating surgeon with a clear knowledge of the anatomy of the second thoracic ganglion during thoracoscopic sympathectomy with a view to improving the success rate for upper limb sympathectomy. PMID- 15800734 TI - Gross morphology of the bridges over the vertebral artery groove on the atlas. AB - The bony bridges of the atlas over the "groove of the vertebral artery" are commonly seen in plain radiographs of the cervical spine, and it is a subject of controversy whether they cause compression of the underneath lying vertebral artery. To clarify this we examined a total of 176 dried and complete atlas vertebrae and found the presence of a "canal for the vertebral artery" (CVA) in 10.23% and an incomplete "canal for the vertebral artery" in 24.43%. The CVA and incomplete CVA is more common in males (11.11% and 24.9%) than in females (9.3% and 24.42%). We found a higher incidence of CVA in laborers (37.5%) than in nonlaborers (4.16%). The incomplete CVA appeared to be more characteristic in the age group of 5-44 years. In the age group of 45-90 years the CVA was characteristic, which probably means that an incomplete CVA is the precursor of a CVA. The superoinferior diameter of the CVA canal ranged from 5.1 to 6.1 mm at the right side and from 4.6 to 5.8 mm at the left side, while the anteroposterior diameter was 5.6-6.9 mm at the right side and 6.1-7.2 mm at the left side. We also found a high incidence of coexistence of CVA and the "retrotransverse foramen" (72.22%) which means that because of possible compression of the vertebral veins the blood flow is directed into the small vein of the retrotransverse foramen. Finally, in 93.5% of unilateral CVA a deeply excavated contralateral "groove of the vertebral artery" was found. PMID- 15800735 TI - Effects of food supplementation on the physiological ecology of female Western diamond-backed rattlesnakes (Crotalus atrox). AB - Food availability is an important factor in the life histories of organisms because it is often limiting and thus can affect growth, mass change, reproduction, and behaviors such as thermoregulation, locomotion, and mating. Experimental studies in natural settings allow researchers to examine the effects of food on these parameters while animals are free to behave naturally. The wide variation among organisms in energy demands and among environmental food resources suggest that responses to changes in food availability may vary among organisms. Since most supplemental feeding field experiments have been conducted on species with high energy demands, we conducted a supplemental feeding study on free-ranging, female Western diamond-backed rattlesnakes (Crotalus atrox), a species with low energy demands and infrequent reproductive investment. Snakes were offered thawed rodents 1-4 times per week. Over two active seasons, we collected data on surface activity, home range size, growth, mass change, and reproduction of supplementally fed and control snakes. Fed and control snakes did not differ in surface activity levels (proportion of time encountered above versus below ground) or home range size. Fed snakes grew and gained mass faster, and had a dramatically higher occurrence of reproduction than control snakes. Also, fed snakes were in better body condition following reproduction than snakes that were not fed. However, litter characteristics such as offspring number and size were not increased by feeding, suggesting that these characteristics may be fixed. These data experimentally demonstrate that food availability can directly impact some life history traits (i.e., growth and reproduction for C. atrox), but not others (i.e., surface activity and home range size for C. atrox). The relationship between food availability and life history traits is affected in a complex way by ecological traits and physiological constraints, and thus interspecific variation in this relationship is likely to be high. PMID- 15800736 TI - Egg mass in an asynchronously hatching parrot: does variation offset constraints imposed by laying order? AB - Life history theory predicts phenotypic trade-offs between the number and quality of offspring produced. Intraspecific variation in egg mass is common in birds and increased egg size can have positive effects on offspring fitness. However, evidence of a trade-off with clutch size is limited. We analyzed variation in mass of 5,743 Green-rumped parrotlet (Forpus passerinus) eggs laid over 15 years to evaluate the potential for facultative adjustment of egg mass and factors governing variation. Heavier eggs had an increased probability of both hatching and fledging but egg mass did not affect postfledging recruitment. Offspring egg mass differed between populations and the potential for seasonal adjustment to egg mass may be related to environmental factors such as seed density. Egg mass was moderately heritable (h2 = 0.42) which accounts for some of the individual variation detected, and these results are likely attributable to strong maternal effects. We found an effect of female age on egg mass, but no effects of previous reproductive experience. Finally, egg mass was strongly governed by position within the laying sequence, independent of clutch size, and such adjustment may facilitate brood reduction under some conditions in this highly asynchronous species. PMID- 15800737 TI - Genotypic variation in tolerance and resistance to fouling in the brown alga Fucus vesiculosus. AB - In this study, we examined genetic variation in resistance and tolerance to fouling organisms in the brown alga Fucus vesiculosus. We first grew 30 algal genotypes in the field, where we allowed fouling organisms to colonise the genotypes at natural levels. We then conducted a manipulative experiment, where we grew 20 genotypes of algae in aquaria with or without fouling organisms. We measured host resistance as the load of fouling organisms and tolerance as the slope of the regression of algal performance on fouling level. Fouling organisms decreased host growth and contents of phlorotannins and thus have the potential to act as selective agents on algal defenses. We found significant among-genotype variation in both resistance and tolerance to fouling. We did not find a trade off between resistance and tolerance. We found a marginally significant cost of resistance, but no cost of tolerance. Our results thus indicate that both the tolerance and resistance of F. vesiculosus can evolve as a response to fouling and that the costs of resistance may maintain genetic variation in resistance. PMID- 15800738 TI - Can greenhouses eliminate the development of cold resistance of the leafminers? AB - Latitudinal patterns for quantitative traits in insect are commonly used to investigate climatic adaptation. We compare the cold resistance of the leafminer (Liriomyza sativae) pupa among populations distributed from tropical to temperate regions, incorporating the thermal overwintering limit of the insect's range. The patterns of cold resistance for northern and southern populations differ. The southern populations significantly increased their cold resistance with latitude, showing a latitudinal pattern independent of seasons, acclimation regimes, and assay methods. In contrast, the northern populations showed no stable patterns; they were always intermediate in cold hardiness between the low-latitude and high latitude populations within the overwintering limit. Integration of these data with those of the biologically similar congeneric leafminer, L. huidobrensis, suggests that a pattern shift in stress tolerance associated with the overwintering range limit is probably a general adaptive strategy adopted by freeze-intolerant species that have a high-latitude boundary of distribution, but can only overwinter and develop in protected greenhouses in harsh seasons. Considering the widespread availability of greenhouses for overwintering insects in northern China, we speculated that the large-scale existence of thermally buffered microhabitats in greenhouses might eliminate the development of cold resistance of the leafminer populations. However, results suggest a strong selection for increased cold resistance for natural populations of Liriomyza species at higher latitudes that can overwinter in the field, but not for populations at latitudes above the thermal limit. Thus, habitat modification associated with greenhouses can limit gene flow and reduce cold tolerances even at latitudes above where the leafminers can overwinter in the field. PMID- 15800739 TI - Water turbidity affects predator-prey interactions in a fish-damselfly system. AB - Community structure may differ dramatically between clear-water and turbid lakes. These differences have been attributed to differences in the cascading effect of fish on prey populations, owing to the reduced efficiency of fish predation in the presence of macrophytes. However, recent theoretical ideas suggest that water turbidity may shape predator-prey interactions, and it is predicted that prey will relax its antipredation behaviour in turbid water (H1). As a result, the nature of predator-prey interactions is expected to shift from both direct and indirect in clear water to dominantly direct in turbid water (H2). We tested these ideas in a fish-damselfly predator-prey system. In a first behavioural experiment, we looked at antipredation behaviour of damselfly larvae isolated from habitats that differ in turbidity, in the presence of fish in clear and turbid water. As predicted in H1, the larvae were more active in turbid than in clear water. In a complementary enclosure experiment, we reared larvae in a clear water pond and a turbid pond, respectively, and manipulated the origin of the larvae (clear-water, turbid pond), fish presence (absent, present), and vegetation density (sparse, abundant). In both ponds, fish had a direct negative effect on survival of the larvae, which was mitigated in the presence of vegetation. In the fish treatment, the change in average body mass tended to be higher in the turbid pond than in the clear-water pond, suggesting indirect effects of fish were mitigated in the turbid pond. This was supported by a negative effect of fish on the effective growth rate of larvae in the clear pond, but not in the turbid pond. These results are compatible with the idea that predator-prey relationships are mainly governed by direct effects in turbid water, and by direct and indirect effects in clear water. PMID- 15800740 TI - Spatio-temporal variability of richness estimators: coastal marine fish communities as examples. AB - We assessed the performance of two estimators of species richness, the Chao2 and the Coleman 'random placement curve'. Using a dataset of intertidal fish from the Norwegian Skagerrak coast, we found that Chao2 was effective for low sampling intensity, often reaching asymptotic values for few samples, but for higher sampling intensity the performance deteriorated. For large samples, the Coleman random placement curve was more effective than the Chao2 estimates when comparing spatio-temporal patterns of species richness. Spatial patterns were clearly and consistently identified by both methods, whereas the coastal fish communities displayed too much variability in the early summer for any sensible measure of temporal patterns of fish-species richness to be made. To control for spurious results due to systematic differences in mean abundance of the samples the analyses were performed also on data standardised by the number of individuals in the samples, without any significant change in the results. We conclude that modest sampling effort is sufficient to characterise spatial patterns of coastal fish-species richness, while a detailed and high-precision description of seasonal patterns could not be obtained with any reasonable sampling effort. PMID- 15800741 TI - Seed weight increases with altitude in the Swiss Alps between related species but not among populations of individual species. AB - Seed weight is a crucial plant life history trait, determining establishment success and dispersal ability. Especially in stressful environments, larger seeds may be selected at the expense of seed number, because larger seeds have a better chance of giving rise to an established offspring. We tested the hypotheses that between related species-pairs and among populations of single species a similar trend for increasing seed weight with increasing altitude should be present. Firstly, we measured seed weights from 29 species-pairs, with one species occurring in lowland areas and a congeneric species from high altitudes. Seeds of the alpine species were 28+/-8% larger than seeds from lowland species (P < 0.01). Compared to the related lowland species, 55% of the alpine species had heavier seeds, 3% (one species) had lighter, and 41% had seeds of approximately equal weight. Secondly, we compared seed weights among populations of four species from different habitats and with different life histories. Seeds from between 11 and 34 populations per species were sampled along altitudinal gradients of 800-1,500 m (ca. 800 m in Scabiosa lucida, ca. 1,000 m in Saxifraga oppositifolia, ca. 1,000 m in Epilobium fleischeri, and ca. 1,500 m in Carex flacca). In all the four species, we found no indication for heavier seeds at higher altitudes. Our results indicate a selection pressure for species with heavier seeds at higher altitude, but the trend does not seem to operate across all cases. Phylogenetic constraints may limit the correlation among altitude and seed weight, operating particularly against selection for larger seed size, the closer populations and species are related to each other. PMID- 15800742 TI - Catching ants with honey: an experimental test of distraction and satiation as alternative modes of escape from flower-damaging ants. AB - According to the distraction hypothesis, extrafloral nectaries (EFN) evolved under selection to entice ants away from floral nectaries, reducing ant-mediated damage to flowers and/or interference with pollinators. Predator-satiation, through production of nectar in either surplus flowers or EFN, provides an alternative mechanism for reducing the impact of ants as flower visitors. I tested these two hypotheses by experimentally adding EFN to flowering plants of the alpine wildflower, Polemonium viscosum, and by surveying the relationship between ant visitation and nectary number in nature. Plants of P. viscosum lack EFN and experience flower damage by ants of Formica neorufibarbus gelida. Ant behavior was compared on plants with five flowers and three experimental EFN and on controls with equal floral display, but no EFN. Addition of EFN increased flower visitation by ants. The effect of EFN on flower visitation did not depend on proximity of EFN to flowers or attractiveness of EFN to ants. Findings suggest that ants perceived patch quality on a whole plant basis, rather than responding to EFN and flowers as distinct nectar patches. Ant visitation did not keep pace with nectary number in nature. The relationship between ant visitation and nectary number per plant was weak and shallow as predicted under satiation. Ant foraging choices on experimental inflorescences showed that ants bypass flowers avoided by earlier ants, enhancing probability of escape via satiation. Results do not support the idea that EFN evolve to reduce flower visitation by ants, but show instead that nectar in surplus flowers can satiate ants and reduce their negative impacts on flower function and integrity. PMID- 15800743 TI - Indirect effects of alternative food resources in an ant-plant interaction. AB - The seeds of many plant species present a food body that is consumed by animal dispersers. In theory, if the animals are polyphagous, the availability of alternative food resource other than the diaspore itself may influence its dispersal and survival. We used the myrmecochore Helleborus foetidus L. (Ranunculaceae), the seeds of which are attached to a lipid-rich elaiosome that is attractive to ants, as a model system to investigate (1) whether alternative foods that are present along with the plant affect ant foraging behavior and diaspore removal and (2) whether food availability in an ant nest affects seed predation and germination. In a field experiment, artificial diaspore depots were offered together with either sugar, insect corpses, seed, or no food (control). Contrary to the prediction that ants would rather concentrate their foraging effort on the highly rewarding alternative foods only, many workers, attracted by the sugar, switched to the hellebore diaspores, which significantly enhanced removal rate. Results obtained in the laboratory further indicated that the larvae of Aphaenogaster iberica (a major seed disperser) predated more on the H. foetidus embryos when no alternative food was available. This, in turn, slightly reduced seed germination. Overall, these results shed light, for the first time, on the potential indirect effects of alternative resources on the fate of diaspores adapted for ant dispersal. PMID- 15800744 TI - Indirect cost of a defensive trait: variation in trichome type affects the natural enemies of herbivorous insects on Datura wrightii. AB - The costs and benefits of defensive traits in plants can have an ecological component that arises from the effect of defenses on the natural enemies of herbivores. We tested if glandular trichomes in Datura wrightii, a trait that confers resistance to several species of herbivorous insects, impose an ecological cost by decreasing rates of predation by the natural enemies of herbivores. For two common herbivores of D. wrightii, Lema daturaphila and Tupiocoris notatus, several generalized species of natural enemies exhibited lower rates of predation on glandular compared to non-glandular plants. Lower rates of predation were associated with reductions in the residence time and foraging efficiency of natural enemies on plants with glandular trichomes, but not with direct toxic effects of glandular exudate. Our results suggest that the benefit of resistance to herbivores conferred by glandular trichomes might be offset by the detrimental effect of this trait on the natural enemies of herbivores, and that the fitness consequences of this trichome defense might depend on the composition and abundance of the natural-enemy community. PMID- 15800745 TI - Amazonia and the modern carbon cycle: lessons learned. AB - In this paper, we review some critical issues regarding carbon cycling in Amazonia, as revealed by several studies conducted in the Large Scale Biosphere Atmosphere Experiment in Amazonia (LBA). We evaluate both the contribution of this magnificent biome for the global net primary productivity/net ecosystem exchange (NPP/NEE) and the feedbacks of climate change on the dynamics of Amazonia. In order to place Amazonia in a global perspective and make the carbon flux obtained through the LBA project comparable with global carbon budgets, we extrapolated NPP/NEE values found by LBA studies to the entire area of the Brazilian Amazon covered by rainforest. The carbon emissions due to land use changes for the tropical regions of the world produced values from 0.96 to 2.4 Pg C year(-1), while atmospheric CO2 inversion models have recently indicated that tropical lands in the Americas could be exchanging a net 0.62+/-1.15 Pg C year( 1) with the atmosphere. The difference calculated from these two methods would imply a local sink of approximately 1.6-1.7 Pg C year(-1), or a source of 0.85 ton C ha(-1) year(-1). Using our crude extrapolation of LBA values for the Amazon forests (5 million km2) we estimate a range for the C flux in the region of -3.0 to 0.75 Pg C year(-1). The exercise here does not account for environmental variability across the region, but it is an important driver for present and future studies linking local process (i.e. nutrient availability, photosynthetic capacity, and so forth) to global and regional dynamic approaches. PMID- 15800746 TI - Stable isotope and band-encounter analyses delineate migratory patterns and catchment areas of white-throated sparrows at a migration monitoring station. AB - The Canadian Migration Monitoring Network consists of several fixed migration monitoring stations (MMS) that apply constant-effort protocols to track changes in the abundance of migratory birds. Such monitoring will be important for tracking long-term population trends of songbirds, especially for species breeding in remote areas such as the North American boreal forest. The geographical catchment sampled by individual MMS, however, remains largely unknown. Here, we used hydrogen isotope measurements (deltaD) of feathers of white-throated sparrows (Zonotrichia albicollis) moving through Delta Marsh MMS in Manitoba, Canada, to determine both wintering and breeding ground catchment areas monitored by this station. The deltaD of tail feathers, collected from spring and fall migrants delineated previous breeding or natal latitudes, ranging from the northern to the southern extremes of the western boreal forest. The deltaD values of head feathers grown on the wintering grounds and collected during spring migration revealed that individuals wintered in a broad region of the southeastern United States. The isotope data showed no relationship between estimated breeding/natal and wintering latitudes of white-throated sparrow populations. Stable isotope data provided little information on longitude. Band encounter analyses, however, indicated a clear east-west segregation of these sparrows across Canada, supporting connectivity among breeding/natal and wintering longitudes over the entire scale of this species' range. Isotope analyses of multiple feather types representing different periods and geographic regions of the annual cycle can provide key information on migratory connectivity for species moving through dedicated MMS. PMID- 15800747 TI - Effects of maternal and embryo characteristics on post-fertilization provisioning in fishes of the genus Gambusia. AB - Maternal provisioning of embryos in Gambusia (Poeciliidae) entails both production of large, yolky eggs and mother-to-embryo transfer of nutrients, the latter of which is readily quantified using injection of radiolabeled nutrients. We assayed patterns of nutrient transfer in broods of 26 Gambusia geiseri and 23 Gambusia affinis females, using injection of tritiated leucine. We examined maternal and embryo characteristics affecting the instantaneous rate of transfer and characterized the pattern of transfer to individual embryos within broods. Maternal (female size and condition) and brood characteristics (mean embryo size, developmental stage, brood size) did not predict the mean level of nutrient transfer to embryos in a brood for either species. Within broods, individual provisioning of embryos was not related to developmental stage, but was related to embryo mass in G. affinis with nutrient transfer higher to larger embryos. In addition, overall within-brood variation in nutrient transfer, measured as coefficient of variation in embryo radioactivity, was higher in G. affinis than in G. geiseri. PMID- 15800748 TI - Chronic herbivory negatively impacts cone and seed production, seed quality and seedling growth of susceptible pinyon pines. AB - Although herbivory often reduces the reproduction of attacked trees, few studies have examined how naturally occurring insect-resistant and susceptible trees differ in their reproduction, nor have these effects been experimentally examined through long-term herbivore removals. In addition, few studies have examined the effects of herbivory on the quality of seeds produced and the implications of reduced seed quality on seedling establishment. We evaluated the impact of chronic herbivory by the stem-boring moth, Dioryctria albovittella, on cone and seed production of the pinyon pine (Pinus edulis) during two mast years. Three patterns emerged. First, moth herbivory was associated with reductions in cone production, viable seed production and seed mass. Specifically, pinyons susceptible to moth attack had 93-95% lower cone production, and surviving cones produced 31-37% fewer viable seeds, resulting in a 96-97% reduction in whole tree viable seed production. In addition, surviving seeds from susceptible trees had 18% lower mass than resistant trees. Second, long-term experimental removal of the herbivore resulted in increased rates of cone and seed production and quality, indicating that moth herbivory was the driver of these reductions. Third, seed size was positively associated with seed germination and seedling biomass and height, suggesting that trees suffering chronic herbivory produce poorer quality offspring. Thus, the resistance traits of pinyons can affect the quality of offspring, which in turn may affect subsequent seedling establishment and population dynamics. PMID- 15800749 TI - Salinity tolerance in Daphnia magna: characteristics of genotypes hatching from mixed sediments. AB - The hatching of diapausing eggs is a means of temporal dispersal that can provide populations with genotypes adapted to different environments. In a salinity variable shallow lake, we predicted that the mixing of different age-classes of eggs in the sediment may yield genotypes with different salinity optima. The alternative would be the absence of local adaptation and the presence of a homogenous population of salt-tolerant genotypes with high phenotypic plasticity. We tested these alternatives by isolating Daphnia magna resting eggs from different sediment depths, exposing them to hatching cues at different salinity levels and measuring the performance of hatched individuals. Results revealed a homogeneous sediment with generally broad-tolerance genotypes and some genotypes with low salt tolerance, which supports the second hypothesis. However, the disturbed character of the sediment hampered historical reconstruction. The absence of local adaptation in the diapausing egg bank may be the result of various scenarios in the response of D. magna populations to severe salinity changes in the lake. PMID- 15800750 TI - Habitat corridors function as both drift fences and movement conduits for dispersing flies. AB - Corridors connect otherwise isolated habitat patches and can direct movement of animals among such patches. In eight experimental landscapes, we tested two hypotheses of how corridors might affect dispersal behavior. The Traditional Corridor hypothesis posits that animals preferentially leave patches via corridors, following them into adjacent patches. The Drift Fence hypothesis posits that animals dispersing through matrix habitat are diverted into patches with corridors because they follow corridors when encountered. House flies (Musca domestica L.), a species that prefers the habitat of our patches and corridors, were released in a central patch (100x100 m) and recaptured in peripheral patches that were or were not connected by a corridor. Flies were captured more frequently in connected than unconnected patches, thereby supporting the Traditional Corridor hypothesis. The Drift Fence hypothesis was also supported, as flies were captured more frequently in unconnected patches with blind (dead end) corridors than in unconnected patches of equal area without blind corridors. A second experiment tested whether these results might be dependent on the type of patch-matrix boundary encountered by dispersing flies and whether edge following behavior might be the mechanism underlying the observed corridor effect in the first experiment. We recorded dispersal patterns of flies released along forest edges with dense undergrowth in the forest ("closed" edges) and along edges with little forest understory ("open" edges). Flies were less likely to cross and more likely to follow closed edges than open edges, indicating that when patch and corridor edges are pronounced, edge-following behavior of flies may direct them along corridors into connected patches. Because edges in the first experiment were open, these results also suggest that corridor effects for flies in that experiment would have been even stronger if the edges around the source patches and corridors had been more closed. Taken together, our results suggest that corridors can affect dispersal of organisms in unappreciated ways (i.e., as drift fences) and that edge type can alter dispersal behavior. PMID- 15800751 TI - The effect of dietary protein quality on nitrogen isotope discrimination in mammals and birds. AB - We tested the competing hypotheses that (1) nitrogen discrimination in mammals and birds increases with dietary nitrogen concentration or decreasing C:N ratios and, therefore, discrimination will increase with trophic level as carnivores ingest more protein than herbivores and omnivores or (2) nitrogen discrimination increases as dietary protein quality decreases and, therefore, discrimination will decrease with trophic level as carnivores ingest higher quality protein than do herbivores. Discrimination factors were summarized for five major diet groupings and 21 different species of birds and mammals. Discrimination did not differ between mammals and birds and decreased as protein quality (expressed as biological value) increased with trophic level (i.e., herbivores to carnivores). Relationships between discrimination factors and dietary nitrogen concentration or C:N ratios were either the opposite of what was hypothesized or non significant. Dietary protein quality accounted for 72% of the variation in discrimination factors across diet groupings. We concluded that protein quality established the baseline for discrimination between dietary groupings, while other variables, such as dietary protein intake relative to animal requirements, created within-group variation. We caution about the care needed in developing studies to understand variation in discrimination and subsequently applying those discrimination factors to estimate assimilated diets of wild animals. PMID- 15800752 TI - The interactive effects of grazing ungulates and aboveground production on grassland diversity. AB - The variable and nonlinear relationships between plant species richness (SR) and aboveground production (NAP) among terrestrial ecosystems indicate that the energetic capacity of ecosystems interacts with other environmental factors to control diversity. One contributing factor determining plant diversity is herbivory; but few studies have effectively examined the interaction of herbivores and NAP on SR. The objective of this study was to investigate how NAP and herds of native migrating ungulates determine plant SR in grasslands of Yellowstone National Park. Plant SR at peak aboveground biomass was compared inside and outside ungulate exclosures at two spatial scales, 1.0 m2 ("local") and 100 m2 ("community"), in ten variable grasslands. NAP also was determined inside and outside exclosures. The relationship between SR and NAP was unimodal for grazed and ungrazed grassland at both spatial scales. Grazers increased local SR, independent of NAP. In contrast, herbivore effects on community SR ranged from no effect among low-productive grassland to an increasingly positive influence as NAP increased. In addition, ungulates reduced beta diversity (the contribution to community SR attributed to variability among local patches) at dry, low-productive and wet, high-productive sites. These results suggest that the size of the pool of species available to colonize grassland is an important factor controlling the response of grassland SR to herbivory, particularly from low- to intermediate-productive grassland. PMID- 15800753 TI - Medium-energy shock wave therapy in the treatment of rotator cuff calcifying tendinitis. AB - To evaluate the results of the treatment with medium-energy extracorporeal shock wave therapy (ESWT) in rotator cuff calcifying tendinitis. Fifty-four non consecutive patients, who were referred to our institute for rotator cuff calcifying tendinitis, were managed with a standardized protocol in four sessions of medium-energy (0.11 mJ/mm2) ESWT administered with an electromagnetic lithotriptor. Pain was evaluated at the end of each session, functional state of shoulder was assessed at 1 and 6 months after the end of procedure. All patients underwent radiographs and sonography imaging. No systemic or local complications. Thirty-eight patients (70%) reported satisfactory functional results. Radiographs and sonographs showed a disappearance of calcium deposit in 29 patients (54%) and in 19 patients (35%) it appeared to be reduced more than a half. A correlation was found between residual calcium deposit and the clinical outcome, but some patients showed a reduced pain without modification of calcium deposit. These results were unmodified at 6 months follow-up. Our protocol of medium-energy ESWT provides good results overall about pain modulation. PMID- 15800754 TI - Time dependent changes in patellar tracking with medial parapatellar and midvastus approaches. AB - The aim of this study was to compare the alteration of patellar tracking by time, which was performed through two different approaches: midvastus and medial parapatellar. Twenty-one patients undergoing simultaneous bilateral primary total knee replacements were randomized prospectively to perform the surgery via a medial parapatellar approach on one knee and midvastus approach on the other knee. All of the patellae were resurfaced. Preoperative, early and late postoperative (mean 22 month) skyline views of the patellae were taken. We found that preoperative 2.24 degrees lateral tilt of patella had not changed by time at the midvastus side (2.95 degrees and 2.57 degrees ). Moreover, preoperative 0.48% lateral subluxation of the patella for midvastus approach had not changed (1.48% and 1.67%). Although 2.19 degrees patellar lateral tilt had not significantly changed at the early postoperative period, which was performed via medial parapatellar approach, there had been a significant increase to 5.38 degrees by time (P=0.037) compared to the preoperative radiographs. Additionally, the preoperative lateral subluxation of the patella (0.57%) at the medial parapatellar side had increased to 5.43% at the early (P=0.009) and 5.62% at the late (P=0.012) postoperative measurements. Midvastus approach is superior to medial parapatellar approach concerning the late patellar tracking. PMID- 15800755 TI - Should we resurface the patella in total knee replacement? Experience with the Nex Gen prothesis. AB - Patellar resurfacing is a controversial issue in primary total knee replacement. So, we took advantage of a large study to try and answer this question. An open prospective multicentric study was conducted about the outcome of the Nex Gen Total Knee prosthesis with clinical and radiological follow-up at regular intervals (1, 2 and 5 years). The data were centrally collected and analyzed by independent observers. A total of whole 5,915 cases were included (487 in the French group); incidences of patello-femoral pain were observed in both French and international groups and compared between cases with or without resurfaced patellae. Only osteoarthritic cases were selected for this particular study. Statistical analysis was performed using a Pearson's Chi-square test with a 95% confidence interval. The frequency of preoperative pain was significantly higher in the French group; 85.1% versus 66.6% (p<0.05). No patello-femoral pain appeared in preoperatively painless patients in the French group. The frequency of lateral patellar release was similar in both groups. 35.7% of the patellae were resurfaced in the international group versus 98.8% in the French one. At 2 years, patello-femoral pain was observed in 6.3% of cases in the former group versus 0.7% in the latter one. The risk of having patello-femoral pain was 9.45 times higher in the international group. When considering resurfaced cases in both groups the risk of having anterior knee pain was not significantly different (p=0.35). In the French group, one case out of nine (11.1%) with patello-femoral pain had not been resurfaced, while 129 cases out of 151 (85.4%) had not had a resurfacing in the international group. This emphasizes the effect of patellar implant on pain. Although patello-femoral pain is multifactorial, this analysis tool is powerful in getting rid of the "center effect " (cf Swedish register). Even if a longer follow-up for confirmation is needed, our data favor patellar resurfacing with this type of implant in spite of the fact that the trochlear design was considered as anatomical. PMID- 15800756 TI - [Allergic and irritant skin and airway diseases in masseurs and medical bath attendants]. AB - The profession of masseur or bath attendant involves intense contact with multiple potential allergens and irritants. There is a complete lack of epidemiological studies for this professional group in spite of the general agreement that sensitization to ingredients of beauty and healthcare products such as fragrances is common. In this review, we critically review the relevant literature, and present own results from recently performed analyses of the German Information Network of Departments of Dermatology (IVDK). Fragrances, preservatives, emulsifiers, and antioxidants are relevant from the allergological perspective. In order to evaluate the particular importance of these and other potentially important allergens for occupational dermatology, further systematic research in allergology as well as epidemiological studies are needed. PMID- 15800757 TI - [Treatment of paraungual HPV73-positive Bowen disease with imiquimod cream]. AB - Bowen disease - a squamous cell carcinoma in situ - is associated with oncogenic human papilloma viruses (HPV). The association is best established for genital Bowen disease but also holds for extragenital lesions. The immunomodulatory substance imiquimod is used for the treatment of HPV-induced skin disorders. In the case of paraungual Bowen disease, established treatment options as excision, cryosurgery, radiotherapy or laser treatment might cause persistent nail damage. We successfully treated HPV73-positive Bowen disease in this location with topical imiquimod. PMID- 15800758 TI - Contribution of flavonoid antioxidants to the preventive effect of mesna in cyclophosphamide-induced cystitis in rats. AB - Cyclophosphamide (CP) is widely used, alone or in combination with other chemotherapeutic agents, for treatment of neoplastic diseases. Its urotoxicity may cause dose-limiting side-effects, for example hemorrhagic cystitis. The agent most often used to prevent this side-effect is mesna (2-mercaptoethane sulfonate). Overproduction of reactive oxygen species during inflammation is one reason for possible urothelial injury. The aim of this study was to evaluate whether combinations of quercetin and epigallocatechin 3-gallate (EGCG), flavonoid antioxidants and mesna could prevent cystitis induced by cyclophosphamide, better than mesna alone. A total of 38 male Sprague-Dawley rats were divided into five groups. Four groups received single dose of CP (100 mg kg( 1)) intraperitoneally at the same time. Group 2 received CP only, group 3 received mesna (3 x 21.5 mg kg(-1)), group 4 received a single dose of mesna+EGCG (2 x 20 mg kg(-1)), and group 5 received a single dose of mesna+quercetin (2 x 20 mg kg(-1)), before and after CP injection. Group 1 (not treated) served as control. CP injection alone resulted in severe cystitis. Mesna resulted in some, but not full, protection against CP toxicity. Quercetin and catechine, together with mesna, resulted in full protection against CP toxicity, on the basis of histopathology of the urinary bladder. It was concluded that oxidants might be important in the pathogenesis of CP-induced cystitis, and that flavonoid antioxidants, used in addition to mesna, may help to ameliorate bladder damage. PMID- 15800762 TI - Chemical vapor generation: are further advances yet possible? PMID- 15800761 TI - Microreactors for drug discovery: the importance of integrating chemical synthesis with real-time analytical detection. PMID- 15800759 TI - Preliminary evaluation of an in utero-lactation assay using 6-n-propyl-2 thiouracil. AB - In this preliminary study, the potential of an in utero-lactation assay to detect thyroid effectors was evaluated by treating three dams/group with 6-n-propyl-2 thiouracil (PTU), a known thyroid antagonist, by oral gavage at doses of 0, 0.0032, 0.016, 0.08 and 0.4 mg/kg/day during fetal organogenesis and lactation. Hearing disturbances and an elevated relative thyroid weight were observed in offspring of both sexes in the 0.4 mg/kg/day group. The Biel-type water T-maze test showed an increase in the number of errors made by females in the 0.4 mg/kg/day group. Histopathologically, flattening of follicular epithelium, a decrease in resorptive colloid droplets, degeneration of follicular epithelium, and hyperplasia of follicular epithelium were observed in males belonging to the 0.4 mg/kg/day group. Histopathological abnormalities were also observed in some offspring belonging to the 0.08 mg/kg/day group. In the dams, hypertrophy of the follicular epithelium of the thyroid was observed in the 0.4 mg/kg/day group. Although we could not explain the mechanism for the difference in the effects seen in the offspring and the dams, the effect of PTU in utero through lactational exposure is apparently different from that resulting from exposure in homeostatically mature rats. Most reports studying PTU have involved administration in water or in food, and reports on the oral gavage of PTU during the fetal organogenesis and lactation period are very rare. This assumes that dosages >0.4 mg/kg/day would also produce clear anti-thyroid effects by oral gavage and, possibly, emphasizes that dosages <0.4 mg/kg/day did not have a noticeable effect. Based on the present results, a study to determine the reproducibility of the data in a much larger number of dams will be performed to confirm the findings in the present study, and to evaluate other endpoints, such as hormonal evaluation of dams and their offspring, sexual developmental landmarks, and fertility of the offspring. PMID- 15800763 TI - Analysis of organic colouring and binding components in colour layer of art works. AB - Two methods of analysis of organic components of colour layers of art works have been tested: IR microspectroscopy of indigo, Cu-phthalocyanine, and Prussian blue, and MALDI-TOF-MS of proteinaceous binders and a protein-containing red dye. The IR spectra distortion common for smooth outer surfaces and polished cross sections of colour layer of art works is suppressed by reflectance measurement of microtome slices. The detection limit of the three blue pigments examined is approximately 0.3 wt% in reference colour layers in linseed oil binder with calcite as extender and lead white as a drying agent. The sensitivity has been sufficient to identify Prussian blue in repaints on a Gothic painting. MALDI-TOF MS has been used to identify proteinaceous binders in two historical paintings, namely isinglass (fish glue) and rabbit glue. MALDI-TOF-MS has also been proposed for identification of an insect red dye, cochineal carmine, according to its specific protein component. The enzymatic cleavage with trypsin before MALDI-TOF MS seems to be a very gentle and specific way of dissolution of the colour layers highly polymerised due to very long aging of old, e.g. medieval, samples. PMID- 15800764 TI - The 18S rDNA sequence of Synchytrium endobioticum and its utility in microarrays for the simultaneous detection of fungal and viral pathogens of potato. AB - Resting spores extracted from wart (Synchytrium endobioticum)-infected potato tubers were used for DNA extraction and amplification of 18S rDNA. Analysis of the cloned, sequenced fragment revealed high similarity to members of the Chytridiomycota. Using this information, specific oligonucleotide probes were designed and arrayed onto glass slides for detection of the pathogen. Viral sequence information available in the databank was retrieved, or new viral sequences were generated, and used to design probes for specific detection of important quarantine viruses of potato. To determine the sensitivity and specificity of the oligonucleotide probes, total RNA from infected plants was reverse transcribed, labelled with Cyanine 5, and hybridised with the microarray. A significant number of the oligonucleotide probes exhibited high specificity to S. endobioticum, Andean potato latent virus, Andean potato mottle virus, Potato black ringspot virus, and Potato spindle tuber viroid. Hybridisation signals of sub-arrays within slides were reproducible (r = 0.79) with a high correlation coefficient of hybridisation repetitions (0.73). Our results demonstrate the potential of microarray-based hybridisation for identification of multiple pathogen targets, which will find application in quarantine laboratories, where parallel testing for diverse pathogens is essential. PMID- 15800766 TI - Antibiotic resistant Streptococcus pneumoniae and hemolytic uremic syndrome. AB - The hemolytic uremic syndrome (HUS) is the most common cause of acute renal failure in young children and most often follows an episode of gastroenteritis caused by an enterohemorrhagic strain of Escherichia coli (O157:H7). HUS induced by Streptococcus pneumoniae (SP) is rare. We report an 18-month-old patient who presented with HUS associated with SP resistant to penicillin and cephalosporins. CONCLUSION: Despite a protracted course including renal failure requiring 15 days of peritoneal dialysis, her kidney function completely recovered. PMID- 15800765 TI - [Correlation of histologic results with PET findings for tumor regression and survival in locally advanced non-small cell lung cancer after neoadjuvant treatment]. AB - CT cannot provide useful information in a timely manner after neoadjuvant treatment. To evaluate the role of (18)F FDG PET after neoadjuvant chemoradiation for early therapy response and its effect on survival as compared to histopathologic tumor response, findings in 32 patients were analyzed prospectively in an ongoing multicenter trial (LUCAS-MD). INCLUSION CRITERIA: histologically confirmed NSCLC stage IIIA/IIIB. Neoadjuvant treatment: 2-3 cycles with paclitaxel/carboplatin and a block of chemoradiation followed by surgery. Pretherapeutic staging: PET scan in addition to a spiral CT and/or MRI. Second PET scan after completion of neoadjuvant therapy prior to surgery. Documentation of lymph node involvement. Assessment of SUV and the metabolic tumor index for primary tumor and metastatic lymph nodes. Image fusion of PET with CT data followed by molecular radiation treatment planning. Evaluation of histologic regression grade and correlation with PET for primary tumor and each lymph node location. All patients (10/32) with complete response in lymph node metastases detected by PET prior to surgery, had no vital tumor cells (i.e. histologic regression grade/RG III, sensitivity 100%). In primary tumors showing complete response, the RG was IIb or III, in one patient IIa (false negative in PET). False positive findings in PET are due to inflammation (5 patients, histologically confirmed). Univariate analyses: actuarial tumor-specific survival for complete metabolic remission vs. incomplete remission after 24 months: 76 vs. 20% (p=0,0079); for RG III/IIb vs. RG IIa/I after 24 months: 63 vs. 36% (p=0,0123).(18)F FDG PET precedes CT in measuring the tumor response and may predict (long term) therapeutic outcome in stage III NSCLC. Histologic regression grade correlates well with metabolic remission as detected by PET. PMID- 15800767 TI - Correlation between "malfunctioning events" and catheter-related infections in pediatric cancer patients bearing tunneled indwelling central venous catheter: results of a prospective observational study. AB - AIM OF THE STUDY: To evaluate the possible link between malfunctioning events and catheter related infections in indwelling central venous devices in children with cancer. PATIENTS AND METHODS: Prospective observation of 418 devices inserted in 2 Italian tertiary care pediatric cancer centers. The presence of a relationship was identified if a malfunctioning event was followed by a catheter related infection within 10 days, or vice versa. RESULTS: The 418 catheters were followed for a period of 107,012 days. Among the malfunctioning events 2 out of 141 (1%) were followed by a catheter related infection while among infectious episodes 3 out of 93 (3%) were followed by a malfunctioning event. CONCLUSIONS: Malfunctioning events followed by catheter related infections and catheter related infections followed by malfunctioning are both rare events in children with cancer. PMID- 15800768 TI - Ending treatment: the course of emotional adjustment and quality of life among breast cancer survivors immediately following radiation therapy. AB - GOAL OF WORK: This study investigated changes in psychological adjustment and quality of life among breast cancer patients following completion of radiation therapy. PATIENTS AND METHODS: Ninety-four patients completed measures of depressed mood, anxiety, and quality of life via interview at five time points: the end of radiation therapy, 2 weeks posttreatment, the first radiation oncology follow-up appointment (4-6 weeks after treatment), 3 months posttreatment, and 6 months posttreatment. MAIN RESULTS: At the conclusion of radiation treatment, participants reported elevated levels of depression, low levels of anxiety, and diminished quality of life. By 2 weeks posttreatment, depression decreased significantly and overall quality of life improved significantly, as well as quality of life in the specific FACT-B domains of Physical and Functional Well Being and the Breast Cancer Subscale. Following that time, the only significant change involved further improvement in breast-cancer-specific concerns. CONCLUSIONS: Results suggest that the primary psychological changes associated with ending breast cancer treatment occur quickly following the conclusion of treatment. Thereafter, psychological status appears to stabilize. The implications of these findings for treatment and directions for future research are discussed. PMID- 15800769 TI - Development of a prognostic index in cancer patients with low performance status. AB - BACKGROUND: Prognosis is a very important medical function. In advanced cancer it is also important to help planning the care to deliver to individual patients with more accuracy, in the process of decision about the opportunity for some interventions and in the selection of patients for clinical trials. Although the performance status indexes are by themselves prognostic factors, among patients in all stages there are wide variations in survival. In what concerns bedridden patients, survival varies between hours and months. Therefore it would be useful to develop a method which could allow a more precise estimation of the length of survival in these patients. PATIENTS AND METHODS: We have studied prospectively 110 bedridden patients exploring six variables: consciousness level, recognition of familiar people, continence and capacity to communicate, to eat and to swallow. Each factor was scored on a scale of 0 to 2. Our aim was to construct a classification system with the sum of the scores of the variables significantly correlated with survival. The cut-off-points were calculated using the percentiles < or =25, 50 and > or =75 according to the method of Altman et al. (J Natl Cancer Inst 86:829-835. 1994). RESULTS: Four of the variables were significantly associated with survival, and an index with three stages was constructed with the sum of these four variables: I-0 to 3; II-4 to 6; III-7 to 8. The differences in survival among the stages are statistically significant and the probability of survival at the 7th, 30th, 60th, and 90th days is also different. CONCLUSION: The differences in survival observed among the stages can be clinically relevant to the establishment of a prognostic to individual patients. PMID- 15800770 TI - Awareness of diagnosis, and information-seeking behavior of hospitalized cancer patients in Greece. AB - PURPOSE: The goal of our study was to evaluate the extent of disease-related knowledge and the need for further information of cancer patients in Greece. MATERIALS AND METHODS: We evaluated 203 mentally competent adult cancer patients hospitalized in general and oncological hospitals in the city of Athens and its suburbs. Data were collected by means of semistructured interviews. Patients were evaluated as to whether they had awareness of their diagnosis. Those who did so (n = 83) were further questioned about additional disease-related information. RESULTS: The majority of patients (59%) claimed to have no knowledge of their diagnosis. Women (p = 0.004) as well as high school and university graduates (p = 0.024) showed significantly superior levels of information when compared to men and graduates of elementary schools, respectively. Age was also a factor that influenced the level of the awareness of the diagnosis and the request for additional information: patients who were informed about the diagnosis and patients who asked for more information were significantly younger than their counterparts who ignored the diagnosis (p < 0.001) and those who didn't ask for further information, respectively (p = 0.03). Hospital specialization (oncological versus general, p < 0.001) and department specialization (medical versus surgical, p = 0.004) were associated with significantly increased levels of information. The patient's educational level was associated with increased request for additional information (p = 0.006). Most patients with knowledge of their diagnosis requested detailed information about severity of their disease and prognosis. Only 13% of those in the informed group claimed they would have benefited psychologically by having been unaware of their diagnosis. CONCLUSIONS: Although Greek patients with diagnoses of malignancies want and need to be adequately informed, the amount of information they receive is inadequate. Over half of those patients evaluated were not aware of their diagnosis. Attitudes of health care professionals, preestablished family beliefs, "mind-set" difficulties, and organizational issues should not become barriers to the patients' right to be fully informed of their diagnoses and choices of potential therapies. PMID- 15800771 TI - Pheromone reception in mammals. AB - Pheromonal communication is the most convenient way to transfer information regarding gender and social status in animals of the same species with the holistic goal of sustaining reproduction. This type of information exchange is based on pheromones, molecules often chemically unrelated, that are contained in body fluids like urine, sweat, specialized exocrine glands, and mucous secretions of genitals. So profound is the relevance of pheromones over the evolutionary process that a specific peripheral organ devoted to their recognition, namely the vomeronasal organ of Jacobson, and a related central pathway arose in most vertebrate species. Although the vomeronasal system is well developed in reptiles and amphibians, most mammals strongly rely on pheromonal communication. Humans use pheromones too; evidence on the existence of a specialized organ for their detection, however, is very elusive indeed. In the present review, we will focus our attention on the behavioral, physiological, and molecular aspects of pheromone detection in mammals. We will discuss the responses to pheromonal stimulation in different animal species, emphasizing the complicacy of this type of communication. In the light of the most recent results, we will also discuss the complex organization of the transduction molecules that underlie pheromone detection and signal transmission from vomeronasal neurons to the higher centers of the brain. Communication is a primary feature of living organisms, allowing the coordination of different behavioral paradigms among individuals. Communication has evolved through a variety of different strategies, and each species refined its own preferred communication medium. From a phylogenetic point of view, the most widespread and ancient way of communication is through chemical signals named pheromones: it occurs in all taxa, from prokaryotes to eukaryotes. The release of specific pheromones into the environment is a sensitive and definite way to send messages to other members of the same species. Therefore, the action of an organism can alter the behavior of another organism, thereby increasing the fitness of either or both. Albeit slow in transmission and not easily modulated, pheromones can travel around objects in the dark and over long distances. In addition, they are emitted when necessary and their biosynthesis is usually economic. In essence, they represent the most efficient tool to refine the pattern of social behaviors and reproductive strategies. PMID- 15800772 TI - The 15th International Chromosome Conference (ICCXV), 5-10 September 2004, Brunel University, UK (Meeting Report). PMID- 15800773 TI - Detection of hepatocellular carcinoma: comparison of Gd-DTPA- and ferumoxides enhanced MR imaging. AB - The aim was to compare the diagnostic performance of dynamic Gd-DTPA- and ferumoxides-enhanced MRI for hepatocellular carcinoma (HCC). Twenty-five patients with chronic hepatitis or liver cirrhosis underwent both dynamic gadopentetate- and ferumoxides-enhanced MRI studies of the liver for HCC detection on the same day. MR data of both studies were retrospectively and independently analyzed. Two observers determined in consensus the grade of diffuse fibrotic liver changes (mild, moderate or severe) and the number of focal lesions. HCCs were confirmed by histology (n=22) and/or follow-up studies for at least six months (n=64). Differences in results obtained from both MR data sets were tested for significance with the McNemar's test (p<0.05). Ferumoxides-enhanced MR images detected 84 of 99 hepatic lesions, including 82 of 86 HCCs and 2 false positive, nonmalignant lesions, while Gd-DTPA-enhanced MR images detected 92 of 99 hepatic lesions, including 81 of 86 HCCs and 11 false positive, nonmalignant lesions. Sensitivity of MRI for detection of HCCs was not significantly different between ferumoxides-enhanced (95.3%; p>0.05) and Gd-DTPA-enhanced scans (94.2%). Gd-DTPA- and ferumoxides-enhanced MRI perform equally well for HCC detection. The majority of small hypervascular hepatic lesions, detected on dynamic Gd-DTPA-enhanced MRI but not on ferumoxides-enhanced MRI, represent no HCCs. PMID- 15800776 TI - [Blood pressure independent effects of antihypertensive agents]. AB - Antihypertensive medications belong to different pharmacological classes. Besides blood pressure lowering properties, many substances, particularly ACE inhibitors and AT1-receptor antagonists but also in part calcium antagonists and aldosterone receptor antagonists, exert additional anti-inflammatory and antifibrotic effects as well as protective effects on endothelium. Delay of disease progression in chronic kidney disorders by inhibition of the renin-angiotensin system also as the result of blood pressure independent effects has been documented in clinical trials. On the other hand, in patients with essential hypertension without end organ damage, it remains unclear whether the clinically proven blood pressure independent effects of antihypertensive agents are also clinically relevant. However, in clinical studies ACE inhibitors and AT1-receptor antagonists reduce the de novo occurrence of the diabetic metabolic state. Inhibition of the renin angiotensin system decreases the incidence of diabetic nephropathy. This contribution presents currently available data on possible blood pressure independent effects of antihypertensive agents. PMID- 15800777 TI - [Alopecia totalis, hypotension and erectile dysfunction in a 34 year old patient. Difficult clarification of a common cause]. AB - The occurrence of both autoimmune endocrinopathies and endocrinopathies caused by other reasons is called polyglandular autoimmune syndrome (PAS-syndrome). In a 34 years old man with weakness, weight loss and erectile dysfunction we found low cortisol caused by an autoimmune adrenalitis and low testosterone caused by a hypophysitis with impaired gonadotropin secretion. Thyroid autoantibodies and islet cell autoantibodies without any hormone deficiencies were further signs of a broad endocrine autoimmune syndrome. In the following 11 years the patient developed three autoimmune disorders: paradrenal glandular insufficiency, hypogonadism caused by hypophysitis, Diabetes mellitus type 1. In the same time several non endocrine autoimmune diseases became manifest: alopecia totalis, vitiligo, retrobulbar neuritis and keratoconjunctivitis. PMID- 15800778 TI - ["A man, who is so dependent on his body as I!". On the 200th anniversary of the death of Friedrich Schiller (1759-1805)]. PMID- 15800779 TI - Agrobacterium-mediated transformation of Phalaenopsis by targeting protocorms at an early stage after germination. AB - A transformation procedure for phalaenopsis orchid established by using immature protocorms for Agrobacterium infection was aimed at the introduction of target genes into individuals with divergent genetic backgrounds. Protocorms obtained after 21 days of culture on liquid New Dogashima medium were inoculated with Agrobacterium strain EHA101(pIG121Hm) harboring both beta-glucuronidase (GUS) and hygromycin resistance genes. Subculture of the protocorms on acetosyringone containing medium 2 days before Agrobacterium inoculation gave the highest transformation efficiencies (1.3-1.9%) based on the frequency of hygromycin resistant plants produced. Surviving protocorms obtained 2 months after Agrobacterium infection on selection medium containing 20 mg l(-1) hygromycin were cut transversely into two pieces before transferring to recovery medium without hygromycin. Protocorm-like bodies (PLBs) proliferated from pieces of protocorms during a 1-month culture on recovery medium followed by transfer to selection medium. Hygromycin-resistant phalaenopsis plants that regenerated after the re-selection culture of PLBs showed histochemical blue staining due to GUS. Transgene integration of the hygromycin-resistant plants was confirmed by Southern blot analysis. A total of 88 transgenic plants, each derived from an independent protocorm, was obtained from ca. 12,500 mature seeds 6 months after infection with Agrobacterium. Due to the convenient protocol for Agrobacterium infection and rapid production of transgenic plants, the present procedure could be utilized to assess expression of transgenes under different genetic backgrounds, and for the molecular breeding of phalaenopsis. PMID- 15800780 TI - Gastrointestinal stromal tumor (GIST) of stomach in a pediatric patient. PMID- 15800781 TI - Functional interaction of CARD15/NOD2 and Crohn's disease-associated TNFalpha polymorphisms. AB - BACKGROUND AND AIMS: Mutations/polymorphisms in the CARD15/NOD2 gene and in the promoter region of the TNFalpha gene are associated with susceptibility to and modulate the phenotype of Crohn's disease (CD). The molecular mechanisms for this genotype-phenotype correlation are yet to be elucidated. CARD15 is an intracellular receptor for bacterial muramyl dipeptide (MDP), and can elicit an inflammatory response via activation of the NF-kappaB pathway. MDP is also known to induce the expression of pro-inflammatory cytokines including TNFalpha, through a still poorly characterized signaling pathway. We sought to determine whether CARD15-mediated NF-kappaB activation can contribute to MDP-induced TNFalpha production and, consequently, if polymorphisms in both genes affect the control of such induction. METHODS/RESULTS: Transfection and electrophoretic mobility shift assays (EMSA) experiments in HEK293 cells demonstrated that MDP exposure stimulates TNFalpha gene transcription, as a result of CARD15-induced NF kappaB activation and binding to TNFalpha promoter. When the CD-associated CARD15 1007fs variant was analyzed, induction of TNFalpha promoter activity was found to be defective. Different combinations of CARD15 and TNFalpha promoter polymorphisms gave rise to distinct TNFalpha transcription levels. CONCLUSIONS: CARD15 and TNFalpha promoter polymorphisms interact to exert a functional effect on MDP-induced TNFalpha production. This gene-gene interaction may contribute to interindividual variation in susceptibility to, and manifestation of, Crohn's disease. PMID- 15800782 TI - Laparoscopic versus open total mesorectal excision: a case-control study. AB - BACKGROUND AND AIMS: Because definitive long-term results are not yet available, the oncological safety of laparoscopic surgery for treatment of rectal cancer remains unproven. The aim of this prospective non-randomised study was to assess the feasibility and short-term outcome of laparoscopic total mesorectal excision (LTME) after 25--30 Gy preoperative radiotherapy and to compare the results with a matched-control group of open TME (OTME). MATERIALS AND METHODS: A series of 41 patients with primary rectal cancer underwent LTME for rectal cancer and were matched with a historical control group of 41 patients who underwent OTME. Both groups received preoperative short-term radiotherapy. RESULTS: There was no mortality in the LTME group and 2% mortality in the OTME group. The overall postoperative morbidity was 37% in the LTME group and 51% in the OTME group, including an anastomotic leakage of 9 and 14% in the LTME and OTME groups respectively. A positive circumferential margin was found in 7% of patients in the LTME group and in 12% of the patients in the OTME group. CONCLUSION: This study shows that LTME is technically feasible and can be performed safely. We show at least a similar surgical completeness using a laparoscopic technique compared with open surgery. PMID- 15800783 TI - [Development of neuroradiology. From the visualization of bones to molecular imaging]. AB - Since the discovery of X-rays, rapid and significant progress has been and continues to be made in imaging techniques, particularly neuroradiology. Milestones along the way included use of contrast agents, digital subtraction angiography, computed tomography, and magnetic resonance imaging. The most recent achievements are visualization of cerebral activation and fiber systems in the brain parenchyma. Application of new contrast agents seems to make imaging at the "molecular" level also possible. PMID- 15800785 TI - Primary effusion lymphoma: successful treatment with highly active antiretroviral therapy and rituximab. PMID- 15800784 TI - Comparison of gray matter and metabolic reduction in mild Alzheimer's disease using FDG-PET and voxel-based morphometric MR studies. AB - PURPOSE: The aim of this study was to investigate regional differences between morphologic and functional changes in the same patients with mild Alzheimer's disease (AD) using statistical parametric mapping (SPM) and voxel-based morphometry (VBM). METHODS: Thirty patients with very mild AD (mean age 66.8 years, mean MMSE score 24.0) and 30 age- and sex-matched normal volunteers underwent both( 18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) and three-dimensional spoiled gradient echo (SPGR) magnetic resonance imaging (MRI). Statistical parametric mapping was used to conduct VBM analysis of the morphological data, which were compared voxel by voxel with the results of a similar analysis of the glucose metabolic data. RESULTS: In AD patients, VBM data indicated a significant gray matter volume density decrease in bilateral amygdala/hippocampus complex (p < 0.05, corrected), while FDG-PET analysis showed significant glucose metabolic reductions in the posterior cingulate gyri and the right parietal lobule, compared with those in the normal control group. CONCLUSION: In very mild AD, morphological change occurs in the medial temporal lobes, while in contrast, metabolic changes occur in the posterior cingulate gyri and parietal lobule. PMID- 15800786 TI - Response rate and survival after thalidomide-based therapy in 248 patients with myelodysplastic syndromes. PMID- 15800787 TI - Remission of transfusion-dependent myelodysplastic syndrome in association with respiratory tract infection. AB - We describe a case of blood transfusion-dependent myelodysplastic syndrome (refractory anaemia), associated with macrocytosis and elevated percentage of hypochromic cells. Following an acute hospital admission with a respiratory tract infection, the patient entered a complete and sustained remission. PMID- 15800788 TI - A gene (pks2) encoding a putative 6-methylsalicylic acid synthase from Glarea lozoyensis. AB - A gene that encodes for a polyketide synthase (PKS) was cloned from the fungus Glarea lozoyensis and characterized. The gene (pks2) consists of four exons interrupted by three introns of 51, 59, and 65 bp, which are clustered at the 5' end. Its predicted product is a 1791-amino-acid protein containing five catalytic motifs typical of fungal PKSs, including a beta-ketosynthase, an acyltransferase, a dehydratase, a beta-ketoacyl reductase, and an acyl carrier region. The gene is transcribed from an initiation site located 375 bp upstream of the translational start codon and extends to a transcriptional termination site 244 bp downstream of the translational stop codon. The gene function is not required for either vegetative growth of G. lozoyensis or for production of pneumocandin, as shown by Agrobacterium-mediated pks2 gene disruption. Previously reported cluster analysis of ketosynthase motifs from 37 fungal polyketide synthases had grouped the Pks2p from G. lozoyensis with PKSs involved in the biosynthesis of 6-methylsalicylic acid. To verify the function of the gene, it was transferred into Aspergillus nidulans under the control of the trpC promoter. 5'-and 3'-RACE experiments confirmed that it was transcribed in the heterologous host, and was associated with the synthesis of a compound identified as 6-methylsalicylic acid by NMR and mass spectrometry. In G. lozoyensis, pks2 is flanked by a gene that encodes a putative drug resistance efflux pump. The Aspergillus pks2 transformants, which were arginine prototrophs, also exhibited precocious pigmentation and accumulated a benzophenone that appeared to be a precursor of emericellin (variecoxanthone B), a known product of A. nidulans. The buildup of the benzophenone may be related to the use of an alternative splice site for the removal of intron 1 of the pks2 transcript in the heterologous host. PMID- 15800789 TI - Management and classification of children with head injury. AB - BACKGROUND: Traumatic brain injury is the most common cause of morbidity and mortality during childhood. AIM: This study was conducted to describe the primary management and classification of children admitted to the emergency department for head trauma. SERIES: All children (0-15 years) with a history of head injury who were admitted to the outpatient emergency department at a single paediatric trauma centre in the Stockholm region during 1 month were included. RESULTS: There were 242 children (137 girls and 105 boys; mean age 5.3+/-4.4 years) who fulfilled the inclusion criteria. Based on the Scandinavian Head Injury Classification, 73% were classified as minimal, 17% mild and 2% moderate head injury. This classification seemed to be more accurately reflective than the ICD 10-based classification in the management of the children with head trauma. PMID- 15800790 TI - Far lateral supracerebellar infratentorial approach for the treatment of upper brainstem gliomas: clinical experience with pediatric patients. AB - OBJECTIVE: Surgical exposure of intrinsic lesions located lateral to the brainstem still represents a challenging task. The aim of this study was to assess the feasibility of the extracerebral far lateral supracerebellar infratentorial (FLSI) approach for the treatment of gliomas located in the upper brainstem in the pediatric population. METHODS AND RESULTS: Between 1992 and 2002, seven patients (mean age 8.7 years) with tumors of glial origin (WHO I-IV) located mainly in the pontomesencephalic region were operated with the FLSI approach in a sitting position. Satisfactory extent of resection without additional morbidity was achieved. CONCLUSION: In a carefully selected pediatric patient population, the FLSI approach proved to be a feasible and effective surgical route for the treatment of upper brainstem gliomas. PMID- 15800791 TI - Rhabdoid meningioma: a new subtype of malignant meningioma also apt to occur in children. AB - CASE REPORT: The case of a 14-year-old girl who presented with a 2-week history of raised intracranial pressure is reported. A left frontal extra-axial tumor was totally removed, whose histopathologic diagnosis was rhabdoid meningioma (RM). DISCUSSION: Rhabdoid meningiomas constitute a special malignant phenotype of meningioma that has been recently included in the WHO classification of tumors of the nervous system. Usually, RMs affect middle-aged and elderly individuals. We report the fourth case of a RM occurring in a child to illustrate that the diagnosis of this tumor subtype, given its prognostic implications, must also be considered in pediatric patients. PMID- 15800792 TI - Intraventricular neurocytoma with massive brain stem involvement in a 5-year-old child. AB - INTRODUCTION: Central neurocytoma is a tumor of neuronal origin that should be taken into consideration in the differential diagnosis of intraventricular neoplasms. Reports of neurocytomas with an extraventricular localization are rare: to our knowledge, the case described here is the first in which a neurocytoma developed within the ventricles but also invaded the brain stem. CASE REPORT: The authors describe the unusual case of a 5-year-old boy with an intraventricular neurocytoma presenting with massive involvement of the basal nuclei and the brain stem. The patient underwent first biopsy and then surgery for ventricular-peritoneal shunting and partial removal of the tumor. Histology showed the tumor to be a typical neurocytoma with Mib-1 <2%. The postoperative course was uneventful. At 3 years' follow-up, the patient's clinical condition is stable and there are no signs of disease progression. DISCUSSION: The literature is reviewed and the characteristics of this unusual tumor are discussed. PMID- 15800794 TI - American Chemical Society--229th National Meeting. PMID- 15800793 TI - Modeling signal and background components of electrosensory scenes. AB - Weakly electric fish are able to detect and localize prey based on microvolt level perturbations in the fish's self-generated electric field. In natural environments, weak prey-related signals are embedded in much stronger electrosensory background noise. To better characterize the signal and background components associated with natural electrolocation tasks, we recorded transdermal voltage modulations in restrained Apteronotus albifrons in response to moving spheres, tail bends, and large nonconducting boundaries. Spherical objects give rise to ipsilateral images with center-surround structure and contralateral images that are weak and diffuse. Tail bends and laterally placed nonconducting boundaries induce relatively strong ipsilateral and contralateral modulations of opposite polarity. We present a computational model of electric field generation and electrosensory image formation that is able to reproduce the key features of these empirically measured signal and background components in a unified framework. The model comprises an array of point sources and sinks distributed along the midline of the fish, which can conform to arbitrary body bends. The model is computationally fast and can be used to estimate the spatiotemporal pattern of activation across the entire electroreceptor array of the fish during natural behaviors. PMID- 15800795 TI - American Chemical Society--229th National Meeting. Oncology. PMID- 15800796 TI - American Chemical Society--229th National Meeting. Kinase modulators. PMID- 15800798 TI - Anti-arthritis therapies 2005. PMID- 15800797 TI - American Chemical Society--229th National Meeting. Oral and poster presentations. PMID- 15800799 TI - Biotechnology--Fourth Croatian International Scientific Conference. Biotechnology and immunomodulatory drugs. PMID- 15800800 TI - Economist conferences 11th Annual Pharmaceuticals Conference. Busting the blocks to growth. PMID- 15800801 TI - Obesity and related disorders--SMi's Second Annual Conference. PMID- 15800802 TI - Retroviruses and opportunistic infections--12th conference. PMID- 15800803 TI - Retroviruses and opportunistic infections--12th conference--HAART. PMID- 15800804 TI - Drug repurposing: identify, develop and commercialize new uses for existing or abandoned drugs. Part I. PMID- 15800805 TI - Drug repurposing: identify, develop and commercialize new uses for existing or abandoned drugs. Part II. PMID- 15800806 TI - MC4 receptor antagonists: a potential treatment for cachexia. AB - Cachexia (involuntary weight loss) is a devastating syndrome associated with many chronic diseases including cancer, and heart, lung, kidney and liver failure. There are currently no effective treatments capable of reversing the loss of lean body mass that is believed to be a major contributor to morbidity and mortality in these chronic diseases. Recent findings strongly indicate that blockade of central melanocortin signaling through the MC4 receptor subtype attenuates cachexia. This review summarizes the evidence supporting the role of MC4 receptors in cachexia, and highlights the progress achieved in the development of small-molecule MC4 antagonists, which have recently proved to be effective in animal models of cachexia. MC4 antagonists are an attractive therapeutic approach for cachexia that may ameliorate the loss of lean body mass in cachectic patients. PMID- 15800807 TI - Treating myelodysplastic syndromes. AB - The myelodysplastic syndromes (MDS) are a diverse group of disorders of hematopoietic stem cells that are characterized by ineffective hematopoiesis, and a variable risk of transformation to acute myeloid leukemia. The prognosis (as estimated by the International Prognostic Scoring System), co-morbidities and age of a patient must all be considered when deciding treatment strategies. Therapeutic modalities vary from supportive care to allogeneic stem cell transplantation (SCT), and growth factors and immunosuppressive therapy may benefit selected patients. The re-emergence of compounds such as thalidomide and arsenic trioxide, and new 'targeted therapy', such as DNA methlytransferase inhibitors and farnesyl tranferase inhibition, provide novel therapeutic options. Furthermore, development of reduced intensity conditioning regimens may result in more patients with MDS benefiting from SCT approaches. PMID- 15800808 TI - Anecortave (Alcon Laboratories). AB - Anecortave, an angiostatic steroid administered as a posterior juxtascleral depot, is under development by Alcon Laboratories Inc for the potential prevention and treatment of ocular diseases, in particular age-related macular degeneration. PMID- 15800809 TI - [Support of psychotherapy research]. PMID- 15800810 TI - [Social anxieties in a clinical sample]. AB - Under the label of social phobia a complex of ailments was introduced into the syndromal diagnostic classification system of ICD-10 and DSM-IV, which is characterized by fear of embarrassing and humiliating experiences in relation to other persons. According to our experiences, however, social phobias and other problems in interpersonal relations are not yet paid the clinical attention that these problems deserve, when the severity of impairments subjects affected by social phobias suffer from is taken into account. There is a considerable lack of research of the psychodynamic aspects of social phobia as well. For this reason, we studied the frequency and extent of social anxieties in a sample of patients requiring psychiatric-psychotherapeutic inpatient treatment suffering from social anxieties (N = 930). This sample was compared to another sample of patients without social anxieties. We applied standardized diagnostic and psychometric instruments in order to collect information about anamnestic and social data as well as data about symptom severity, interpersonal impairments and comorbid disorders. The results do not only show the high frequency of social anxieties (45 %), but also the severe impairments in various domains of personal functioning of patients suffering from social anxieties compared to the patients without severe social anxieties. According to our clinical experiences and to the results presented here, the construct of social phobia as it is currently used in the psychiatric classification systems is an syndromal additive description that falls short of a form of suffering that is characterized by the facts that it is interwoven with the complete social existence of an affected person. For this reason, the term social phobia should be replaced by the more appropriate term of social anxiety. PMID- 15800811 TI - [Psychotherapy service utilization and psychotherapy motivation in a representative community sample of the elderly -- results of the zurich older age study]. AB - OBJECTIVE: To study the extent of real and potential psychotherapy service utilization in the older (65 - 96) population. METHOD: We used the Zurich Older Age Study on PTSD, complicated grief and depressive disorders which consists of a random sample of 712 older adults based on a two-stage screening design. Utilization of psychotherapy was assessed by a checklist on service utilization for psychological problems/disorders. Potential psychotherapy motivation was assessed by means of a validated Questionnaire on Psychotherapy Motivation (FPTM 23). RESULTS: Around 5 % indicated to have used psychotherapy since age 65 (either alone or in combination with pharmacological treatment) and 10 % of participants indicated use of any therapeutic treatment for psychological problems with women twice as frequent as men. Potential psychotherapy motivation was medium highly pronounced and varied only slightly between the sexes. Predictors of high psychotherapy motivation are the extent of suffering as well as the absence of rejection of psychological need for assistance. DISCUSSION: The frequency numbers of psychotherapy service utilization and motivation indicate their moderate extents in old age. Limitations of the study are discussed, e. g., the possibly higher density of service providers in an urban population like the city of Zurich. PMID- 15800812 TI - [Consensus paper on the terminological differentiation of various aspect of body experience]. AB - In the past, phenomenological research on subjective body experience was characterised by vaguely defined terminology and methodological shortcomings. The term "body image" has been applied heterogeneously in literature in order to describe a variety of bodily phenomena. In this paper, the German terminology applied to the phenomenology of body experiences is described systematically. In developing a systematic terminology the authors refer to scientific evidence as well as recent reviews, and closely adhere to definitions commonly used in English literature. Different perspectives are utilised, particularly anthropological concepts and theories from developmental and self-psychology. Distinct aspects of body experience are described within the context of a network of external determinants and along a continuum between somatic and mental anchor points. Applying the term "body experience" as umbrella term, different aspects are defined: perceptive (body schema/-perceive), affective (body-cathexis), cognitive-evaluative (body-image, body-ego) and body-consciousness. It is emphasized, that the distinct description of functional levels has to be taken as an approximation of the reality of integrated body experience. PMID- 15800813 TI - [Assessing psychosomatic aspects of chronic orofacial pain syndrome]. AB - BACKGROUND: The diagnosis of COS is often formulated when classification fails for the lack of clear symptoms or persisting complaints in spite of evidence based dental care. PATIENTS AND METHODS: Eight patients suffering from COS and referred to us by the Dental Clinic of the Munich University were invited to complete the self-rating SASB-Intrex questionnaire. In addition, the patients were examined in a clinical psychodynamic interview. RESULTS: All patients suffered from undefined, very often bizarre, complaints of the teeth, oral cavity, and face. These dysesthetic syndromes occurred after previous dental interventions. However, the complaints surpass symptomatic pain due to local aetiologies or neurological causes. There was clinical evidence of underlying unconscious conflicts, e. g., concerning control vs. submission. All patients showed clinically-relevant psychosomatic co-morbidity which could be formulated as cyclic maladaptive interpersonal patterns. CONCLUSION: COS patients are often not sufficiently examined and undergo multiple, sometimes irreversible and invalidating dental or surgical interventions. Given their exclusively somatic attribution style, referral to psychotherapy is frequently unsuccessful. Characteristic cyclic maladaptive interaction patterns may contribute to the chronification process. This study shows the difficulties and chances of an interdisciplinary diagnosis and treatment program. PMID- 15800814 TI - [The Kansas City Cardiomyopathy Questionnaire (KCCQ) -- a new disease-specific quality of life measure for patients with chronic heart failure]. AB - Patients' health-related quality of life is increasingly being included as an additional endpoint when evaluating the treatment of chronic heart failure. Although generic self-report instruments measuring health-related quality of life are available, there is a lack of disease-specific instruments covering various dimensions of quality of life with high reliability, validity and sensitivity to chance. Thus, the aim of the present study was to evaluate the German version of a new heart failure-specific quality of life measure, the Kansas City Cardiomyopathy Questionnaire (KCCQ). The sample consisted of 233 consecutively recruited outpatients of a university department in Germany. Test-retest reliability was high (intraclass correlation coefficient 0.93 for both the Functional State and the Clinical Summary total scores). Construct validity was demonstrated with strong correlations to respective subscales of the SF-36. Known groups validity was shown by both statistically and clinically significant differences between NYHA classes. The examination of sensitivity to change yielded promising results. The questionnaire was well accepted by the participating patients. The KCCQ proved to be a reliable and valid self-report instrument for measuring disease-specific quality of life in chronic heart failure. PMID- 15800815 TI - [The Inventory "State-Trait Operation Anxiety" (STOA): construction and empirical findings]. AB - This article reports on the construction and empirical evaluation of an instrument for the measurement of surgery-related state and trait anxiety. The inventory "State-Trait Operation Anxiety" (STOA) separately assesses surgery related anxiety as a comparatively stable personality trait as well as the cognitive and affective components of state anxiety. Results of explorative and confirmatory factor analyses corroborated the unifactorial structure of trait anxiety and the two-factorial structure of state anxiety. Internal consistencies of all scales were highly satisfactory (Cronbach's alpha around 0.90). External relationships of the STOA with trait anxiety, dispositional and actual coping, observed anxiety, as well as indicators of perioperative adjustment confirmed the inventory's validity. An analysis of the scores of the two state anxiety components across the perioperative period demonstrated that these scales are sensitive indicators of state anxiety changes. PMID- 15800816 TI - [How useful is the calculation of total scores for questionnaires concerning health related quality of life?]. AB - Questionnaires for health related quality of life generally comprise several subscales. These subscales can be summarized into a total score. The aim of this article is to test the justification of such an integration from a statistical point of view. By means of a representative sample of the German population (n = 1981) the correlative relationships among six questionnaires are assessed: NHP, EORTC QLQ-C30, HADS, MFI-20, GBB-24 and Whiteley index. The corrrelations between the total scores are markedly higher than the mean correlations among the subscales between the tests. Even in those cases where the subscales share the same subdimension the correlations are not higher than the correlations between these subscales and the (unspecific) total scores. The advantage of sharing a common area is as great as the disadvantage of the lower number of items. By means of the Spearman-Brown formula the relationship between the number of items and the reliability of the scale is discussed in detail. It can be concluded that the calculation of total scores is useful and justified from a statistical perspective. PMID- 15800817 TI - [The dream of flying]. AB - More than a 100 years ago the Wright brothers succeeded in performing the first motor flight in the history of mankind. But irrespective of its technical realisation man has always dealt with flying. So myths, rites and fairy-tales as well reflect the different ideas of flying as these conceptions come to light again and again in dreams and visions. Whether ascension, expression of desire and yearning or sexual metaphor -- the idea of flying seems to be a universal magic figure of thinking. PMID- 15800818 TI - [Against a morality of indifference towards suffering]. PMID- 15800819 TI - [Prevention of colorectal carcinoma: a conditio sine qua non!]. PMID- 15800820 TI - [Comparison of different stool tests for the detection of cancer of the colon]. AB - BACKGROUND AND OBJECTIVE: Colonoscopy is the gold standard for the diagnosis of colonic neoplasia. Because of the low compliance, the discomfort of bowel preparation and the procedure itself and the (albeit small) risk of perforation or bleeding alternative procedures such as stool tests are being focused on. PATIENTS AND METHODS: After informed consent stool samples of 116 patients (44 male, 72 female, median age 47 years), scheduled for colonoscopy and 22 patients (17 m, 5 f, 69 y) with known colorectal cancer stool samples were collected. The samples were investigated by three methods: a biochemical (Guajak) test for fecal occult blood, an immunological test for fecal occult blood and a test determining the dimer pyruvat kinase M2 (tumor M2-PK). RESULTS: Sensitivity for detection of colorectal cancer or polyps was 27 % and 10 % for the biochemical test, 91 % and 19 % for the immunological test and 77 % and 48 % for the M2-PK-test, respectively. Specificity was 89 %, 94 % and 72 %, respectively. CONCLUSIONS: Both methods for detection of occult blood had a similar specificity. The sensitivity of the immunological test for the detection of colorectal cancer was significantly higher. The M2-PK-test had a markedly lower specificity in diagnosing cancer. Because of the low sensitivity for polyps the usefulness of stool tests is questionable. Reducing incidence and mortality of colorectal cancer should be achieved by colonoscopy, a recommendation that requires specific communication to the public. PMID- 15800821 TI - [Codeine-induced pancreatitis]. PMID- 15800822 TI - [Colon cancer: molecular markers]. PMID- 15800823 TI - [A new method for the treatment of depression: repetitive transcranial magnetic stimulation]. AB - Recent data suggest that repetitive transcranial magnetic stimulation (rTMS) is effective in treating depressive symptoms to a lesser extent compared with classical electroconvulsive therapy. However, rTMS represents an economical and well tolerable procedure in relation to the expenditure of electroconvulsive therapy with anaesthesia. Usually, rTMS is applicated as an add-on-therapy accompanying psychopharmacological treatment. So far, it has predominantly been used for patients with long-standing and so called treatment-refractory symptoms. However, even in the early phase of a depressive episode rTMS would be possibly more effective. In many cases, the standard procedure-application of up to 10 rTMS-sessions will not be enough to produce therapeutic benefit. Therefore rTMS series including up to 20 sessions are recommended. Long-term studies are needed to clarify the role of rTMS for relapse prevention and to determine the optimal frequency and duration of rTMS in such an indication. Although numerous results of newer studies suggest a moderate antidepressive effect of rTMS, its application in daily clinical routine practice cannot be recommended yet. Larger, accurate designed and controlled studies, especially involving patients of old age, are needed to evaluate the true tolerability and effectiveness of rTMS as a new treatment option for depressive symptoms. PMID- 15800824 TI - [Obesity and cardiovascular morbidity]. PMID- 15800825 TI - [Integrity and autonomy at the end of life]. PMID- 15800826 TI - [Duty of the hospital representative to appoint attending physicians -- decision of Labor Court Wilhelmshaven, 9/23/2004]. PMID- 15800827 TI - [Proinsulin as marker -- what for?]. PMID- 15800828 TI - [Postmyocardiotomy syndrome]. PMID- 15800829 TI - [Stress processing in medicine -- concerning the articles in DMV 3/2005]. PMID- 15800834 TI - [Osteoporosis in patients under oral anticoagulant treatment. Concerning the article in DMW 50/2004]. PMID- 15800835 TI - [San Antonio Breast Cancer Symposium -- highlights 2004]. AB - The adjuvant therapy of postmenopausal, hormonereceptor positive breast cancer patients with aromatase inhibitors in ATAC, ABCSG 8/ ARNO 95 and IES trial revealed just a benefit in terms of disease-free survival, but not for over-all survival. Thus MA-17 trial is so far the only study which could demonstrate a survival benefit for node positive disease by "extended adjuvant" therapy with letrozole. Node- and receptorpositive, postmenopausal women show a higher benefit by anthracycline containing chemotherapy in combination with tamoxifen, than by endocrine therapy alone. But tamoxifen should given sequentially to chemotherapy (Intergroup 0100/ SWOG-8814 trial). Adjuvant chemotherapy with three cycles FEC followed by three cycles docetaxel reveals a survival benefit compared to six cycles FEC, especially in women older than 50 years (PACS 01 trial). Furthermore there is evolving evidence that dose-dense, interval shortened chemotherapy is superior to standard chemotherapy in adjuvant setting. The impaired survival of receptornegative, nodalpositive breast cancer patients seems to be converted to a survival improvement compared with receptorpositive patients by modern adjuvant cytotoxic regimens (metaanalysis CALGB 8541, 9344, 9741). In primary systemic chemotherapy survival is strongly associated with the rate of pathohistological complete remissions (NSABP B-27). The combination of cytotoxic drugs has no further influence on survival, but the sequential administration of taxane increases the rate of pCR. A difference in survival stratified by pCR and non-pCR in breast and positive nodal status could not be detected, so that the number of involved lymph nodes seems to be the strongest prognostic factor and is independent of tumor response in the breast. The sentinel node biopsy is a safe and reliable method, but the rate of false negative findings depends on the pathohistological methods (NSABP B-32). The morbidity of this method is low compared with conventional axillary dissection in multicenter studies. Local tumor control is an important parameter which offers a significant survival benefit in all age cohorts. PMID- 15800836 TI - [Forensic aspects of female breast reconstructive surgery]. PMID- 15800837 TI - [Treatment of breast cancer patients with brain metastases]. AB - Treatment of patients with brain metastases is based on an interdisciplinary approach. It is essential to perform a differenciated indication for procedures with consideration of all therapeutical options like modalities of radiotherapy, surgery and oncology. In the case of multiple brain metastases the whole brain radiotherapy is the standard of treatment, while in case of a single or solitary brain metastasis surgical procedures followed by radiotherapy should be preferred. PMID- 15800838 TI - [Magnetic resonance imaging (MRI) of the pelvis in diagnosing endometriosis]. AB - Widely accepted as an established method in the diagnostic imaging of diseases of the female pelvis, magnetic resonance imaging (MRI) holds promising potential for the diagnosis of endometriosis. This review describes the general prerequisites for performing MRI scans of the pelvis and presents, based on recent studies, an imaging protocol suited to the diagnosis of endometriosis. Typical manifestations and sites of endometriosis are discussed and illustrated with sample images. For endometriotic cysts and implants, axial T (1)-weighted, fat-saturated turbo spin echo sequences (TSE) have the highest diagnostic value. In diagnosing adenomyosis, sagittal T (2)-weighted TSE sequences visualise changes in the uterine zonal anatomy. In essence, MRI is capable of detecting many manifestations of endometriosis in a non-invasive, yet reliable manner, and its value in relation to laparoscopy must be considered as complementary. PMID- 15800839 TI - [Prevention of preeclampsia]. AB - Preeclampsia is accompanied by high fetal and maternal morbidity and mortality and to a high degree responsible for preterm delivery. The pathophysiologic mechanisms underlying this disease remain poorly understood. Accordingly, only few causative or preventive therapeutical strategies are known. One such example for a preventive strategy is the use of aspirin (ASA) which directly affects the imbalance of vasodilative prostacycline and vasoconstrictive thromboxane. Recently, some studies are indicating a preventive effect of vitamin C and E substitution. In contrast, early antihypertensive therapy did not prevent later progression of the disease. Furthermore, sodium restriction, calcium and magnesium substitution, fish oil substitution, or steroid therapy are without any effect regarding the later development of preeclampsia. It is of utmost importance to further elucidate underlying pathophysiological mechanisms to improve therapeutical and preventive strategies. PMID- 15800840 TI - [Anaesthetic-related complications of caesarean section]. AB - In spite of an increase in the number of caesarean sections, the incidence of anaesthetic-related complications remains low. This is due primarily to the increasing use of regional anaesthesia (spinal and or epidural anaesthesia) as general anaesthesia is associated with a 17-fold increase in complications, in particular failed endotracheal intubation, aspiration of gastric contents and hypoxia. It is most important that all obstetric patients deemed at risk for general anaesthesia (e. g. morbidly obese, hypertension, placenta praevia) should be identified as such by the obstetricians and referred to the anaesthetic department at an early stage. Furthermore there should be common procedural guidelines to define the degree of urgency of caesarean section as most anaesthetic complications occur during "emergency" cases under general anaesthesia. Many of these so-called emergency cases are not real emergencies and could be equally well performed under regional anaesthesia. A continuing audit of maternal deaths in Germany should be established along similar lines to the Confidential Enquiries into Maternal Deaths in the United Kingdom. Most importantly, the increasing use of regional anaesthesia should be propagated as its use in Germany still lies well behind other countries such as Switzerland, the USA and the United Kingdom. PMID- 15800841 TI - [Vaginal foreign body -- cause for a rectovaginal fistula]. AB - A case of 56-year-old patient is presented with the diagnosis "carcinoma of the cervix". She suffered from brown, bad smelling vaginal discharge since half a year without any pain. Three weeks before she had noticed a postmenopausal bleeding. At the examination under anaesthesia with cysto- and rectoscopy we saw a rectovaginal fistula. In the vagina a white plastic object was found which could not be removed. Some days later a hysterectomy with extraction of the foreign body (aerosol cap) was done and the fistula was treated together with the surgeons. There was no evidence of a carcinoma. The patient suffers from multiple sclerosis since 20 years. On asking she told us that the foreign body was in place since about two years. She was not willing to relate any other information. PMID- 15800842 TI - [Severe virilization in a postmenopausal woman: a case report]. AB - Severe postmenopausal virilization is a rare event in clinical practice. To evaluate ovarian or adrenal hyperandrogenism endocrine tests and imaging are useful diagnostic tools. We report a case of a postmenopausal woman with hirsutism and androgenetic alopecia. A malignant cause for the present disorder could be excluded by imaging. Selective venous sampling was administered with increased testosterone level of the right adrenal vein. Right adrenalectomy and right oophorctomy was recommended. PMID- 15800843 TI - A novel STX16 deletion in autosomal dominant pseudohypoparathyroidism type Ib redefines the boundaries of a cis-acting imprinting control element of GNAS. AB - A unique heterozygous 3-kb microdeletion within STX16, a closely linked gene centromeric of GNAS, was previously identified in multiple unrelated kindreds as a cause of autosomal dominant pseudohypoparathyroidism type Ib (AD-PHP-Ib). We now report a novel heterozygous 4.4-kb microdeletion in a large kindred with AD PHP-Ib. Affected individuals from this kindred share an epigenetic defect that is indistinguishable from that observed in patients with AD-PHP-Ib who carry the 3 kb microdeletion in the STX16 region (i.e., an isolated loss of methylation at GNAS exon A/B). The novel 4.4-kb microdeletion overlaps with the previously identified deletion by 1,286 bp and, similar to the latter deletion, removes several exons of STX16 (encoding syntaxin-16). Because these microdeletions lead to AD-PHP-Ib only after maternal transmission, we analyzed expression of this gene in lymphoblastoid cells of affected individuals with the 3-kb or the 4.4-kb microdeletion, an individual with a NESP55 deletion, and a healthy control. We found that STX16 mRNA was expressed in all cases from both parental alleles. Thus, STX16 is apparently not imprinted, and a loss-of-function mutation in one allele is therefore unlikely to be responsible for this disorder. Instead, the region of overlap between the two microdeletions likely harbors a cis-acting imprinting control element that is necessary for establishing and/or maintaining methylation at GNAS exon A/B, thus allowing normal G alpha(s) expression in the proximal renal tubules. In the presence of either of the two microdeletions, parathyroid hormone resistance appears to develop over time, as documented in an affected individual who was diagnosed at birth with the 4.4-kb deletion of STX16 and who had normal serum parathyroid hormone levels until the age of 21 mo. PMID- 15800844 TI - Over- and underdosage of SOX3 is associated with infundibular hypoplasia and hypopituitarism. AB - Duplications of Xq26-27 have been implicated in the etiology of X-linked hypopituitarism associated with mental retardation (MR). Additionally, an expansion of a polyalanine tract (by 11 alanines) within the transcription factor SOX3 (Xq27.1) has been reported in patients with growth hormone deficiency and variable learning difficulties. We report a submicroscopic duplication of Xq27.1, the smallest reported to date (685.6 kb), in two siblings with variable hypopituitarism, callosal abnormalities, anterior pituitary hypoplasia (APH), an ectopic posterior pituitary (EPP), and an absent infundibulum. This duplication contains SOX3 and sequences corresponding to two transcripts of unknown function; only Sox3 is expressed in the infundibulum in mice. Next, we identified a novel seven-alanine expansion within a polyalanine tract in SOX3 in a family with panhypopituitarism in three male siblings with an absent infundibulum, severe APH, and EPP. This mutation led to reduced transcriptional activity, with impaired nuclear localization of the mutant protein. We also identified a novel polymorphism (A43T) in SOX3 in another child with hypopituitarism. In contrast to findings in previous studies, there was no evidence of MR or learning difficulties in our patients. We conclude that both over- and underdosage of SOX3 are associated with similar phenotypes, consisting of infundibular hypoplasia and hypopituitarism but not necessarily MR. PMID- 15800847 TI - Epithelial cell polarity genes are required for neural tube closure. AB - Human neural tube defects (NTD) are a heterogeneous group that exhibit complex inheritance, making it difficult to identify the underlying cause. Due to the uniform genetic background, inbred mouse strains are a more amenable target for genetic studies. We investigated the loop-tail (Lp) mouse as a model for the severe NTD, craniorachischisis. A homozygous point mutation was identified in the transmembrane protein Vangl2, which in Drosophila has been shown to function in the planar cell polarity (PCP) pathway. Morphological analysis of the Lp mice shows that the defect results from an abnormally broad floor plate, most likely through a failure in convergent extension. The elevated neural folds remain too far apart to contact, inhibiting neural tube closure. Recently, two other mouse mutants (crash and circletail) were described with a similar phenotype to Lp and were investigated as potentially new alleles. Mapping studies, however, showed that both mutants segregated to distinct loci. In the crash (Crsh) mouse, a mutation was identified in Celsr1, a seven pass transmembrane receptor that encodes a protein orthologous to Drosophila Flamingo. Like Vangl2, this gene also functions in the PCP pathway. While in circletail, a point mutation was identified introducing a premature stop codon into the apical-basal cell polarity gene scribble (Scrb1). We subsequently demonstrated a genetic interaction between all three genes, where double heterozygotes exhibit the same homozygous NTD phenotype. This strongly suggests both a candidate gene pathway and that interaction between independent recessive alleles may be a possible explanation for the complex inheritance in severe human NTD. PMID- 15800845 TI - Quantitative founder-effect analysis of French Canadian families identifies specific loci contributing to metabolic phenotypes of hypertension. AB - The Saguenay-Lac St-Jean population of Quebec is relatively isolated and has genealogical records dating to the 17th-century French founders. In 120 extended families with at least one sib pair affected with early-onset hypertension and/or dyslipidemia, we analyzed the genetic determinants of hypertension and related cardiovascular and metabolic conditions. Variance-components linkage analysis revealed 46 loci after 100,000 permutations. The most prominent clusters of overlapping quantitative-trait loci were on chromosomes 1 and 3, a finding supported by principal-components and bivariate analyses. These genetic determinants were further tested by classifying families by use of LOD score density analysis for each measured phenotype at every 5 cM. Our study showed the founder effect over several generations and classes of living individuals. This quantitative genealogical approach supports the notion of the ancestral causality of traits uniquely present and inherited in distinct family classes. With the founder effect, traits determined within population subsets are measurably and quantitatively transmitted through generational lineage, with a precise component contributing to phenotypic variance. These methods should accelerate the uncovering of causal haplotypes in complex diseases such as hypertension and metabolic syndrome. PMID- 15800846 TI - Microduplication and triplication of 22q11.2: a highly variable syndrome. AB - 22q11.2 microduplications of a 3-Mb region surrounded by low-copy repeats should be, theoretically, as frequent as the deletions of this region; however, few microduplications have been reported. We show that the phenotype of these patients with microduplications is extremely diverse, ranging from normal to behavioral abnormalities to multiple defects, only some of which are reminiscent of the 22q11.2 deletion syndrome. This diversity will make ascertainment difficult and will necessitate a rapid-screening method. We demonstrate the utility of four different screening methods. Although all the screening techniques give unique information, the efficiency of real-time polymerase chain reaction allowed the discovery of two 22q11.2 microduplications in a series of 275 females who tested negative for fragile X syndrome, thus widening the phenotypic diversity. Ascertainment of the fragile X-negative cohort was twice that of the cohort screened for the 22q11.2 deletion. We also report the first patient with a 22q11.2 triplication and show that this patient's mother carries a 22q11.2 microduplication. We strongly recommend that other family members of patients with 22q11.2 microduplications also be tested, since we found several phenotypically normal parents who were carriers of the chromosomal abnormality. PMID- 15800850 TI - Understand and use your rights. PMID- 15800849 TI - Technical note: a useful guide catheter modification for anomalous right coronary arteries. PMID- 15800852 TI - Mouse models of neural tube defects: investigating preventive mechanisms. AB - Neural tube defects (NTD), including anencephaly and spina bifida, are a group of severe congenital abnormalities in which the future brain and/or spinal cord fail to close. In mice, NTD may result from genetic mutations or knockouts, or from exposure to teratogenic agents, several of which are known risk factors in humans. Among the many mouse NTD models that have been identified to date, a number have been tested for possible primary prevention of NTD by exogenous agents, such as folic acid. In genetic NTD models such as Cart1, splotch, Cited2, and crooked tail, and NTD induced by teratogens including valproic acid and fumonisins, the incidence of defects is reduced by maternal folic acid supplementation. These folate-responsive models provide an opportunity to investigate the possible mechanisms underlying prevention of NTD by folic acid in humans. In another group of mouse models, that includes curly tail, axial defects, and the Ephrin-A5 knockout, NTD are not preventable by folic acid, reflecting the situation in humans in which a subset of NTD appear resistant to folic acid therapy. In this group of mutants alternative preventive agents, including inositol and methionine, have been shown to be effective. Overall, the data from mouse models suggests that a broad-based in utero therapy may offer scope for prevention of a greater proportion of NTD than is currently possible. PMID- 15800851 TI - Role of Folbp1 in the regional regulation of apoptosis and cell proliferation in the developing neural tube and craniofacies. AB - Folic acid is essential for many cellular reactions, including synthesis of nucleotides and regulation of cell cycle. Folic acid-binding protein one (Folbp1), a membrane-bounded protein, is the primary mediator of folic acid transport. Mice deficient in Folbp1 gene die in utero with multiple malformations, including severe exencephaly and craniofacial defects. Fusion of the neural tube and craniofacies require precisely regulated interactions of apoptosis, cell proliferation, and differentiation. To understand the role of Folbp1 in regulating the fusions of these primordia, levels of dead and proliferating precursor cells from Folbp1 embryos were quantified before the fusion processes. Massive apoptosis was detected in the Folbp1-/- defective tissues, with Bax and activated caspase-3 distributed evenly across the apico basal axis of the lateral neural plate. 5-Bromodeoxyuridine (BrdU) and PCNA labeling assays revealed a reduced cell proliferation as well. However, telomerase activity was unaltered, arguing against telomere shortening and consequently, chromosomal instability, as the cause of the apoptosis. Notably, Islet-1 and 2H3 immunohistochemistry demonstrated the presence of differentiating neuronal cells, albeit in decreased numbers. Interestingly, Folbp1-/- embryos also elaborated novel neural structures that sprouted orthogonally from the embryonic neuraxis. Assays on the defective craniofacies exhibited similar phenomena, suggesting the neural crest precursor population that gives rise to both these structures is selectively vulnerable to Folbp1 inactivation. The results demonstrate a prominent role of Folbp1 in the regional regulation of apoptosis and cell proliferation that underlies the aberrant neural tube and craniofacial defects. PMID- 15800853 TI - Current perspectives on the causes of neural tube defects resulting from diabetic pregnancy. AB - Maternal diabetes increases the risk for neural tube, and other, structural defects. The mother may have either type 1 or type 2 diabetes, but the diabetes must be existing at the earliest stages of pregnancy, during which organogenesis occurs. Abnormally high glucose levels in maternal blood, which leads to increased glucose transport to the embryo, is responsible for the teratogenic effects of maternal diabetes. Consequently, expression of genes that control essential developmental processes is disturbed. In this review, some of the biochemical pathways by which excess glucose metabolism disturbs neural tube formation are discussed. Research from the author's laboratory has shown that expression of Pax3, a gene required for neural tube closure, is significantly reduced by maternal diabetes, and this is associated with significantly increased neural tube defects (NTD). Pax3 encodes a transcription factor that has recently been shown to inhibit p53-dependent apoptosis. Evidence in support of this model, in which excess glucose metabolism inhibits expression of Pax3, thereby derepressing p53-dependent apoptosis of neuroepithelium and leading to NTD will be discussed. PMID- 15800854 TI - Non-multifactorial neural tube defects. AB - Although most neural tube defects (anencephaly, spina bifida) occur as isolated malformations, a substantial proportion are attributable to chromosome anomalies, known teratogens, or component manifestations of multiple anomaly syndromes. This review describes known chromosome alterations and the candidate genes residing in the altered region, as well as syndromes associated with neural tube defects and causative genes, if known. PMID- 15800855 TI - Osteocalcin enhances bone remodeling around hydroxyapatite/collagen composites. AB - The effect of osteocalcin (OC), an extracellular bone matrix protein, on bone healing around hydroxyapatite/collagen composites was investigated. Cylindrical nanocrystalline hydroxyapatite implants of 2.5-mm diameter containing 2.5% biomimetically mineralized collagen type I were inserted press-fit into the tibial head of adult Wistar rats. To one implant group, 10 mug/g OC was added. Six specimens per group were analyzed at 2, 7, 14, 28, and 56 days. After 14 days, newly formed woven bone had reached the implant surface of the OC implants whereas a broad fibrous interface could still be observed around controls. Woven bone was formed directly around both implant groups after 28 days and had been replaced partially by lamellar bone around the OC implants only. No significant differences in total bone contact were seen between both groups after 56 days. The higher number of phagocytosing cells and osteoclasts characterized immunohistochemically with ED1, cathepsin D, and tartate-resistant alkaline phosphatase around the OC implants at the early stages of bone healing suggests an earlier onset of bone remodeling. The earlier and increased expression of bone specific matrix proteins and multifunctional adhesion proteins (osteopontin, bone sialoprotein, CD44) at the interface around the OC implants indicates that OC may accelerate bone formation and regeneration. This study supports the observations from in vitro studies that OC activates both osteoclasts and osteoblasts during early bone formation. PMID- 15800856 TI - Refractive index measurement in viable cells using quantitative phase-amplitude microscopy and confocal microscopy. AB - BACKGROUND: The refractive index (RI) of cellular material provides fundamental biophysical information about the composition and organizational structure of cells. Efforts to describe the refractive properties of cells have been significantly impeded by the experimental difficulties encountered in measuring viable cell RI. In this report we describe a procedure for the application of quantitative phase microscopy in conjunction with confocal microscopy to measure the RI of a cultured muscle cell specimen. METHODS: The experimental strategy involved calculation of cell thickness by using confocal optical sectioning procedures, construction of a phase map of the same cell using quantitative phase microscopy, and selection of cellular regions of interest to solve for the cell RI. RESULTS: Mean cell thickness and phase values for six cell regions (five cytoplasmic and one nuclear) were determined. The average refractive index calculated for cytoplasmic and nuclear regions was 1.360 +/- 0.004. The uncertainty in the final RI value represents the technique measurement error. CONCLUSIONS: The methodology we describe for viable cell RI measurement with this prototype cell has broad generic application in the study of cell growth and functional responses. The RI value we report may be used in optical analyses of cultured cell structure and morphology. PMID- 15800857 TI - Cytotoxic capacity of hepatitis C virus (HCV)--specific lymphocytes after in vitro immunization with HCV-derived lipopeptides. AB - BACKGROUND: Hepatitis C virus (HCV)-derived lipopeptides can induce epitope specific immune responses in lymphocytes from HCV-naive individuals. We analyzed whether such T cells generated by in vitro immunization with HCV core-derived lipopeptides exert HCV-specific cytolytic activity. METHODS: Using a sensitive flow cytometric cytotoxicity assay we characterized HCV-specific cytotoxicity in T cells generated in vitro with HCV core-derived 25-mer lipopeptides. In addition, we studied expressions of Fas ligand and perforin and interferon-gamma (IFN-gamma) secretion in HLA-A2-HCV(core_35-44) tetramer-positive T cells generated with lipopeptide amino acid 20-44 (LP20-44). RESULTS: CD8+ T cells induced in vitro with HCV core-derived lipopeptides only infrequently exerted HCV specific cytotoxicity, irrespective of whether antigen-coated T2 cells or autologous B lymphoblasts were used as targets. Detailed analysis of HLA-A2 HCV(core_35-44) tetramer-positive T cells generated with LP20-44 revealed that in vitro immunization resulted in T cells that secreted IFN-gamma after antigen specific restimulation and that upregulated expression of Fas ligand but not of perforin. CONCLUSIONS: Our data confirm at the functional level that HCV lipopeptides induce antigen-specific T lymphocytes that produce IFN-gamma but exert significant cytotoxicity in only a minority of experiments, probably because expression of cytolytic effector molecules is not enhanced in their granules. PMID- 15800858 TI - Efficiency of magnetic liposomal transforming growth factor-beta 1 in the repair of articular cartilage defects in a rabbit model. AB - We evaluated the efficacy of a magnetic liposomal delivery system of transforming growth factor (TGF)-beta(1) in the treatment of articular cartilage defects in a rabbit model. Articular cartilage defects were created in the patellar groove of rabbits, and a permanent magnet or a nonmagnetic alloy was implanted in the defect site. Magnetic liposomal drugs, prepared by the conventional film method and sonication, were injected into the defect site 1 week after surgery. First, the efficacy of the magnetic liposomal delivery system was evaluated by using a model compound fluorescence-labeled dextran 40,000 (FD-40). Then, the therapeutic efficiency of magnetic liposomal TGF-beta(1) was evaluated by cartilage histological scoring at 4, 8, and 12 weeks after surgery. The injected magnetic liposomal FD-40 accumulated at the target site where a permanent magnet had been implanted. The histological score showed that the injection of magnetic liposomal TGF-beta(1) under magnetic force was significantly effective in the repair of the defect site over 12 weeks after surgery. Injection of TGF-beta(1) into the cartilage defect was effective as a magnetic liposomal preparation under magnetic force, resulting in acceleration of the cartilage repair, probably because of the desirable accumulation of TGF-beta(1) at the target site. PMID- 15800860 TI - Fluorene and phenanthrene uptake by Pseudomonas putida ATCC 17514: kinetics and physiological aspects. AB - Pseudomonas putida ATCC 17514 was used as a model strain to investigate the characteristics of bacterial growth in the presence of solid fluorene and phenanthrene. Despite the lower water-solubility of phenanthrene, P. putida degraded this polycyclic aromatic hydrocarbon (PAH) at a maximum observed rate of 1.4 +/- 0.1 mg L(-1) h(-1), higher than the apparent degradation rate of fluorene, 0.8 +/- 0.07 mg L(-1) h(-1). The role of physiological processes on the biodegradation of these PAHs was analyzed and two different uptake strategies were identified. Zeta potential measurements revealed that phenanthrene-grown cells were slightly more negatively charged (-57.5 +/- 4.7 mV) than fluorene grown cells (-51.6 +/- 4.9 mV), but much more negatively charged than glucose grown cells (-26.8 +/- 3.3 mV), suggesting that the PAH substrate induced modifications on the physical properties of bacterial surfaces. Furthermore, protein-to-exopolysaccharide ratios detected during bacterial growth on phenanthrene were typical of biofilms developed under physicochemical stress conditions, caused by the presence of sparingly water-soluble chemicals as the sole carbon and energy source for growth, the maximum value for TP/EPS during growth on phenanthrene (1.9) being lower than the one obtained with fluorene (5.5). Finally, confocal laser microscopy observations using a gfp-labeled derivative strain revealed that, in the presence of phenanthrene, P. putida::gfp cells formed a biofilm on accessible crystal surfaces, whereas in the presence of fluorene the strain grew randomly between the crystal clusters. The results showed that P. putida was able to overcome the lower aqueous solubility of phenanthrene by adhering to the solid PAH throughout the production of extracellular polymeric substances, thus promoting the availability and uptake of such a hydrophobic compound. PMID- 15800861 TI - Temperature-triggered purification of antibodies. AB - In this article the unique capability of elastin-like protein (ELP) to reversibly precipitate was combined with the high affinity and specificity of antibody binding domains such as Protein G, Protein L, or Protein LG as a general method for antibody purification that combines in a unique manner the simplicity and robustness of temperature-triggered precipitation with the selectivity of affinity interactions. In a single precipitation step, antibodies derived from different sources (animal sera or hybridoma cell cultures) were selectively recovered by a simple temperature trigger. Due to the versatility of the binding ligands toward different classes of antibodies, we believe that this technology will be useful as an economical, highly efficient, and universal platform for the purification of antibodies. PMID- 15800862 TI - Membrane fouling in a membrane bioreactor (MBR): sludge cake formation and fouling characteristics. AB - A submerged membrane bioreactor (MBR) with a working volume of 1.4 L and a hollow fiber microfiltration membrane was used to treat a contaminated raw water supply at a short hydraulic retention time (HRT) of approximately 1 h. Filtration flux tests were conducted regularly on the membrane to determine various fouling resistances, and confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were employed to characterize the biofouling development and sludge cake formation on the membrane. The experimental results demonstrate that the MBR is highly effective in drinking water treatment for the removal of organic pollutants, ammonia, and UV absorbance. During the MBR operation, the fouling materials were not uniformly distributed on the entire surface of all of the membrane fibers. The membrane was covered partially by a static sludge cake that could not be removed by the shear force of aeration, and partially by a thin sludge film that was frequently washed away by aeration turbulence. The filtration resistance coefficients were 308.4 x 10(11) m(-1) on average for the sludge cake, 32.5 x 10(11) m(-1) on average for the dynamic sludge film, and increased from 10.5 x 10(11) to 59.7 x 10(11) m(-1) for the membrane pore fouling after 10 weeks of MBR operation at a filtration flux of 0.5 m3/m2 x d. Polysaccharides and other biopolymers were found to accumulate on the membrane, and hence decreased membrane permeability. More important, the adsorption of biopolymers on the membrane modified its surface property and led to easier biomass attachment and tighter sludge cake deposition, which resulted in a progressive sludge cake growth and serious membrane fouling. The sludge cake coverage on the membrane can be minimized by the separation, with adequate space, of the membrane filters, to which sufficient aeration turbulence can then be applied. PMID- 15800863 TI - Gene delivery through cell culture substrate adsorbed DNA complexes. AB - Efficient gene delivery is a fundamental goal of biotechnology and has numerous applications in both basic and applied science. Substrate-mediated delivery and reverse transfection enhance gene transfer by increasing the concentration of DNA in the cellular microenvironment through immobilizing a plasmid to a cell culture substrate prior to cell seeding. In this report, we examine gene delivery of plasmids that were complexed with cationic polymers (polyplexes) or lipids (lipoplexes) and subsequently immobilized to cell culture or biomaterial substrates by adsorption. Polyplexes and lipoplexes were adsorbed to either tissue culture polystyrene or serum-adsorbed tissue culture polystyrene. The quantity of DNA immobilized increased with time of exposure, and the deposition rate and final amount deposited depended upon the properties of the substrate and complex. For polyplexes, serum modification enhanced reporter gene expression up to 1500-fold relative to unmodified substrates and yielded equivalent or greater expression compared to bolus delivery. For lipoplexes, serum modification significantly increased the number of transfected cells relative to unmodified substrates yet provided similar levels of expression. Immobilized complexes transfect primary cells with improved cellular viability relative to bolus delivery. Finally, this substrate-mediated delivery approach was extended to a widely used biomaterial, poly(lactide-co-glycolide). Immobilization of DNA complexes to tissue culture polystyrene substrates can be a useful tool for enhancing gene delivery for in vitro studies. Additionally, adapting this system to biomaterials may facilitate application to fields such as tissue engineering. PMID- 15800865 TI - Serum albumin-catalyzed trigger system by using a tandem kemp elimination/beta elimination reaction. PMID- 15800864 TI - Biomass characteristics and membrane aeration: toward a better understanding of membrane fouling in submerged membrane bioreactors (MBRs). AB - Fouling in submerged membrane bioreactors (MBRs) was studied under different operating conditions and with varying biomass characteristics. Fouling rates were determined using a flux-step method for seven biomass conditions with mixed liquor solids concentrations ranging from 4.3 to 13.5 g x l(-1), six permeate fluxes (5.5, 11.0, 16.5, 22.0, 27.5, and 33.0 l x m(-2) x h(-1)), and three membrane airflow velocities (0.07, 0.10, and 0.13 m x s(-1)). Statistical analysis was used to specify the degree of influence of each of the biomass characteristics (solids concentration, dewaterability, viscosity, particle size distribution, concentrations of protein and carbohydrate in the soluble microbial products, SMP, and extracellular polymer substances, EPS), the permeate flux and the membrane aeration velocity on the membrane fouling rate. Among all these variables, only the permeate flux, the solids concentration (correlated to the viscosity and the dewaterability), the carbohydrate concentration in the EPS, and the membrane aeration velocity were found to affect the fouling rate. The permeate flux had the greatest effect. A transitional permeate flux was observed between 16.5 and 33 l x m(-2) x h(-1), below which no significant fouling was observed regardless of the biomass characteristics, the permeate flux, and the membrane aeration velocity. PMID- 15800866 TI - Potent and selective inhibition of class II alpha-D-mannosidase activity by a bicyclic sulfonium salt. PMID- 15800867 TI - Beyond oil and gas: the methanol economy. PMID- 15800868 TI - The intermolecular Pauson-Khand reaction. AB - Five membered carbocycles are important building blocks for many biologically active molecules. Moreover, substituted cyclopentenones (e.g. cyclopentenone prostaglandins) exhibit characteristic biological activity. The efficiency and atom economy of the Pauson-Khand reaction render this process potentially one of the most attractive methods for the synthesis of such compounds. Although it was discovered in its intermolecular form, the scope of the intermolecular Pauson Khand reaction has always been limited by the poor reactivity and selectivity of the alkene component. The past decade, especially the last three years, has seen concerted efforts to broaden the scope of this reaction. In this overview, we provide a comprehensive and critical coverage of the intermolecular Pauson-Khand reaction based on the reactivity characteristics of different classes of alkenes and a rationalization of successes and misfortunes in this area. PMID- 15800869 TI - An exceptionally selective lead(II)-regulatory protein from Ralstonia metallidurans: development of a fluorescent lead(II) probe. PMID- 15800870 TI - Reversible binding of dioxygen by a non-transition-metal complex. PMID- 15800871 TI - Incarcerated carbenes. PMID- 15800872 TI - Changes in the proteomic profile during differentiation and maturation of human monocyte-derived dendritic cells stimulated with granulocyte macrophage colony stimulating factor/interleukin-4 and lipopolysaccharide. AB - Dendritic cells (DCs) are highly specialized antigen-presenting cells that play an essential role in the immune response. We used the proteomic approach based on two-dimensional gel electrophoresis and mass spectrometry to identify the protein changes that occur during differentiation of DCs from monocytes (Mo) stimulated with granulocyte macrophage colony stimulating factor/interleukin-4 (GM-CSF/IL-4) and during the maturation of immature DCs stimulated with lipopolysaccharide. Sixty-three differentially expressed proteins (+/- two-fold) were unambiguously identified with sequence coverage greater than 20%. They corresponded to only 36 different proteins, because 11 were present as 38 electrophoretic forms. Some proteins such as tropomyosin 4 and heat shock protein 71 presented differentially expressed electrophoretic forms, suggesting that many of the changes in protein expression that accompany differentiation and maturation of DCs occur in post translationally modified proteins. The largest differences in expression were observed for actin (21-fold in Mo), Rho GDP-dissociation inhibitor 2 (20-fold in Mo), vimentin (eight-fold in immature DCs), lymphocyte-specific protein 1 (12 fold in mature DCs) and thioredoxin (14-fold in mature DCs). Several proteins are directly related to functional and morphological characteristics of DCs, such as cytoskeletal proteins (cytoskeleton rearrangement) and chaperones (antigen processing and presentation), but other proteins have not been assigned specific functions in DCs. Only a few proteins identified here were the same as those reported in proteomic studies of DCs, which used different stimuli to produce the cells (GM-CSF/IL-4 and tumor necrosis factor-alpha). These data suggest that the DC protein profile depends on the stimuli used for differentiation and especially for maturation. PMID- 15800873 TI - Proteomics of ischemia/reperfusion injury in rabbit myocardium reveals alterations to proteins of essential functional systems. AB - Brief periods of myocardial ischemia prior to timely reperfusion result in prolonged, yet reversible, contractile dysfunction of the myocardium, or "myocardial stunning". It has been hypothesized that the delayed recovery of contractile function in stunned myocardium reflects damage to one or a few key sarcomeric proteins. However, damage to such proteins does not explain observed physiological alterations to myocardial oxygen consumption and ATP requirements observed following myocardial stunning, and therefore the impact of alterations to additional functional groups is unresolved. We utilized two-dimensional gel electrophoresis and mass spectrometry to identify changes to the protein profiles in whole cell, cytosolic- and myofilament-enriched subcellular fractions from isolated, perfused rabbit hearts following 15 min or 60 min low-flow (1 mL/min) ischemia. Comparative gel analysis revealed 53 protein spot differences (> 1.5 fold difference in visible abundance) in reperfused myocardium. The majority of changes were observed to proteins from four functional groups: (i) the sarcomere and cytoskeleton, notably myosin light chain-2 and troponin C; (ii) redox regulation, in particular several components of the NADH ubiquinone oxidoreductase complex; (iii) energy metabolism, encompassing creatine kinase; and (iv) the stress response. Protein differences appeared to be the result of isoelectric point shifts most probably resulting from chemical modifications, and molecular mass shifts resulting from proteolytic or physical fragmentation. This is consistent with our hypothesis that the time course for the onset of injury associated with myocardial stunning is too brief to be mediated by large changes to gene/protein expression, but rather that more subtle, rapid and potentially transient changes are occurring to the proteome. The physical manifestation of stunned myocardium is therefore the likely result of the summed functional impairment resulting from these multiple changes, rather than a result of damage to a single key protein. PMID- 15800874 TI - Proteomic analysis of Lyme disease: global protein comparison of three strains of Borrelia burgdorferi. AB - The Borrelia burgdorferi spirochete is the causative agent of Lyme disease, the most common tick-borne disease in the United States. It has been studied extensively to help understand its pathogenicity of infection and how it can persist in different mammalian hosts. We report the proteomic analysis of the archetype B. burgdorferi B31 strain and two other strains (ND40, and JD-1) having different Borrelia pathotypes using strong cation exchange fractionation of proteolytic peptides followed by high-resolution, reversed phase capillary liquid chromatography coupled with ion trap tandem mass spectrometric analysis. Protein identification was facilitated by the availability of the complete B31 genome sequence. A total of 665 Borrelia proteins were identified representing approximately 38% coverage of the theoretical B31 proteome. A significant overlap was observed between the identified proteins in direct comparisons between any two strains (>72%), but distinct differences were observed among identified hypothetical and outer membrane proteins of the three strains. Such a concurrent proteomic overview of three Borrelia strains based upon only the B31 genome sequence is shown to provide significant insights into the presence or absence of specific proteins and a broad overall comparison among strains. PMID- 15800877 TI - Epidemiology of neural tube defects. AB - The epidemiological investigation of the common open neural tube defects (NTDs), anencephaly, and spina bifida, has a long history. The most significant finding from these past studies of NTDs was the identification of the protective effect of maternal, periconceptional supplementation with folic acid. Fortuitously, the association between folic acid and NTDs became widely accepted in the early 1990s, at a time when genetic association studies of complex traits were becoming increasingly feasible. The confluence of these events has had a major impact on the direction of epidemiological, NTD research. Association studies to evaluate genes that may influence the risk of NTDs through their role in folate-related processes, or through other metabolic or developmental pathways are now commonplace. Moreover, the study of genetic as well as non-genetic, factors that may influence NTD risk through effects on the nutrient status of the mother or embryo has emerged as a major research focus. Research efforts over the past decade indicate that gene-gene, gene-environment, and higher-order interactions, as well as maternal genetic effects influence NTD risk, highlighting the complexity of the factors that underlie these conditions. The challenge for the future is to design studies that address these complexities, and are adequately powered to detect the factors or combination of factors that influence the development of NTDs. PMID- 15800878 TI - Rapid progression of pericardial calcification in a patient with end-stage renal disease. AB - Dialysis pericarditis is a relatively uncommon cause of pericardial constriction and may be found in patients with end-stage renal disease receiving adequate renal replacement therapy. We present a patient with end-stage renal disease maintained on chronic peritoneal dialysis who developed severe myopericardial calcification over a 2-month period demonstrated by sequential chest computed tomographic scanning. The characteristic hemodynamic findings of constrictive effusive pericarditis, obtained during cardiac catheterization, are presented and discussed. PMID- 15800879 TI - Improved detection of minimal acute myeloid leukemia cells by the use of the combined parameters of XE-2100 hematology analyzer. AB - BACKGROUND: For the diagnosis and therapy of acute leukemia, it is important to detect a small number of leukemic cells precisely. Although several automated hematology analyzers that carry blast-detecting programs have been developed, they do not exert sufficient detection sensitivity to exceed the sensitivity of manual eye counting method. METHODS: We constructed a new blast-detecting program by combining the numerical information acquired from five cytometric parameters presented by XE-2100. The sensitivity and specificity of this blast multi-scoring program were assessed in comparison with the Blasts flag program equipped originally in XE-2100. RESULTS: The blast-detecting sensitivity was found to be highly improved in the blast multi-scoring program as compared with the Blasts flag program without much decreasing the specificity. A small number of leukemic myeloblasts was detected at the better sensitivity than the eye counting method in the clinical course of the patients with acute myeloid leukemia. CONCLUSIONS: The daily practical use of this blast multi-scoring program will surely contribute to sensitive, objective, and real-time evaluation of the control of acute myeloid leukemia with a low cost. PMID- 15800880 TI - Serum parathyroid hormone-related peptide is associated with systemic inflammation and adverse prognosis in gastroesophageal carcinoma. AB - BACKGROUND: Parathyroid hormone-related peptide (PTHrP) is a tumor-derived circulating factor that has been associated with hypercalcemia of malignancy. The role of PTHrP as a prognostic indicator remains unclear. Studies suggest that it may function as a growth factor; and, recently, the ability of PTHrP to induce cytokine expression has been described. PTHrP also has been proposed as a procachectic factor. In this study, the authors investigated the prognostic value of PTHrP in patients who had gastroesophageal carcinoma without hypercalcemia and determined whether PTHrP was associated with systemic inflammation and adverse nutritional status. METHODS: Patients were recruited at the time of diagnosis. Serum was collected for determination of c-terminal fragment PTHrP (cPTHrP) levels (by radioimmunoassay) and calcium levels as well as levels of serum cytokines and acute-phase proteins (with an enzyme-linked immunosorbent assay). Nutritional assessment of patients was undertaken at the same time as serum collection. Patients underwent routine staging, and survival duration was recorded. RESULTS: One hundred fifty-one patients with esophagogastric carcinoma were recruited. Six of 151 patients (4.0%) patients were hypercalcemic, and 26 patients (17.2%) had elevated serum cPTHrP levels. There was no association between the cPTHrP level and either serum calcium concentrations (P = 0.72) or adverse nutritional status. Elevated cPTHrP, however, was associated with significantly higher serum levels of soluble tumor necrosis factor receptor (P = 0.008) and with significantly lower levels of transferrin (P = 0.009) and albumin (P = 0.02). There was also a weak association with C-related protein levels (P = 0.06). Elevated cPTHrP levels also were associated with an adverse prognosis, as determined by reduced survival duration, on univariate analysis (P = 0.038), but not on multivariate analysis (P = 0.15). CONCLUSIONS: Elevated serum cPTHrP levels were present in approximately 17% of patients with gastroesophageal carcinoma in the absence of hypercalcemia and was associated with markers of systemic inflammation and with an adverse prognosis. PMID- 15800881 TI - NOR expression increases on metaphase chromosomes of Down syndrome lymphocytes in concordance with mitogen concentration in culture medium. AB - BACKGROUND: Regulation of nucleolus organizer region (NOR) expression in trisomy 21 (Down syndrome [DS]) cells is not fully explained. This work compared NOR expression on metaphase chromosomes in gradiently stimulated lymphocytes from DS patients with those from healthy controls. METHOD: Conventional peripheral blood culture (72 h) and chromosomal preparation procedures were used except that blood samples from each individual were cultivated in the same but gradiently increasing concentrations (0.37, 0.75, 1.48, and 2.21 ml) of phytohemagglutinin (PHA) per 100 ml of medium. One hundred consecutive metaphases per concentration were analyzed for scoring the means of the active NORs bearing chromosomes (AgNOR+ chromosome) per individual and per concentration. RESULTS: In contrast to healthy controls (n=24), AgNOR+ chromosomal number in lymphocytes from 30 DS patients increased in concordance to the gradient of PHA concentration in the culture medium. CONCLUSION: DS lymphocytes do not downregulate their NOR expression in the limit of control cells. This in vitro result may serve as a clue for the explanation of the DS phenotype due to the wasted energy in producing unnecessary rRNA transcripts and AgNOR proteins in utero during organogenesis. These results also indicate that precautions must be used in routine work of NOR evaluation/interpretation in DS lymphocytes. PMID- 15800882 TI - Impact of preanalytical variables on granulocytic surface antigen expression: a review. AB - BACKGROUND: There is a gradual but steady increase in the use of granulocytic surface marker studies to diagnose several inherited and acquired blood and bone marrow disorders. Diagnosis and follow-up of patients with inflammation and infection are other areas of quantitative flow cytometric application. Despite the increased use of flow cytometry to study granulocytes, there seems to be no well-established standards regarding specimen handling for these studies. METHODS: This review summarizes the effect of preanalytical variables on granulocytic surface markers. RESULTS AND CONCLUSIONS: Storing specimens in sodium heparin at room temperature for up to 72 h seems satisfactory. Other anticoagulants, although acceptable, may have a shorter storage time. Storage time could be prolonged further with the use of some preservation media. Lysed whole blood is the preferred technique. Techniques should avoid major temperature change and excessive manipulation and should maintain storage and methodologic temperature with minimal fluctuation. Fixation before staining with the antibody may result in decreased expression of some surface antigens. PMID- 15800883 TI - A multicenter evaluation of the PanLeucogating method and the use of generic monoclonal antibody reagents for CD4 enumeration in HIV-infected patients in Thailand. AB - BACKGROUND: The current method of CD4 enumeration in Thailand, based on the three tube, three-color method recommended by the Centers for Disease Control and Prevention, is expensive and thus unavailable to most patients who have the human immunodeficiency virus (HIV). Less expensive, simpler protocols (i.e., PanLeucogating and primary CD4 gating) have been described but require more published validation data to gain widespread acceptance. We describe a multicenter evaluation of the PanLeucogating method. METHODS: The PanLeucogating method using generic reagents was evaluated in comparison with the standard three tube, three-color method using commercial reagents. Percentage of CD4+ T cells among lymphocytes and absolute CD4+ T-cell counts were determined in 611 HIV infected individuals recruited from four sites. Linear regression and Bland Altman tests were used for statistical analysis. RESULTS: The correlation of percentage of CD4+ T cells and absolute CD4+ T-cell counts obtained with the PanLeucogating strategy and the standard predicate method was high (r2 = 0.96 and 0.95, respectively, for the entire study population and r2 > 0.95 and 0.93, respectively, for each study group). Absolute CD4+ T-cell counts of the overall study pool and of the two subdivisions of absolute CD4+ T-cell counts (i.e., 0 250 cells/microl and > 250 cells/microl) derived from the two methods demonstrated excellent agreement, with mean biases of +18 cells/microl, +11 cells/microl, and +24 cells/microl, respectively. CONCLUSIONS: These observations demonstrate that CD4 enumeration by PanLeucogating is reliable and can be performed to an identical standard in a quality-assured network of collaborating laboratories as a new cost-effective approach to HIV monitoring. PMID- 15800884 TI - Pexiganan-incorporated collagen matrices for infected wound-healing processes in rat. AB - The use of peptide-based drugs is limited by their rapid degradability and toxicity at high concentration during their therapeutic application. These problems could be managed by the use of a peptide delivery agent for sustained release in the site of action. Collagen is one of the most proven biomaterials of good biocompatibility with an exceptional ligand encapsulating property. In this work, we have shown that pexiganan, an antimicrobial, 22-amino-acid peptide could be incorporated and delivered to the wound-healing site against bacterial strains Pseudomonas aeruginosa and Staphylococcus aureus. The release profiles of pexiganan collagen films with different collagen concentration were studied. The release of pexiganan from 2.5% w/w of collagen film showed a sustainable activity over 72 h with effective antimicrobial concentrations. Pexiganan-incorporated collagen (PIC)-treated groups were compared with open wound (OW)- and collagen film (CF)-treated rats. PIC-treated animals showed a diminishing level of bacterial growth as compared with OW- and CF-treated animals. The biochemical parameters such as hydroxyproline, protein, DNA, uronic acid, hexosamine, SOD, and catalase content in the granulation tissue of the healing wound revealed increased proliferation of cells involved in tissue reconstruction in PIC-treated groups when compared with OW- and CF-treated groups. Furthermore, spectroscopic studies suggested that collagen structure is not perturbed by pexiganan incorporation. This study provides rationale for application of collagen membrane for antimicrobial peptide delivery in infected wounds. PMID- 15800886 TI - Pilomyxoid astrocytoma in a patient with neurofibromatosis. AB - Pilomyxoid astrocytoma (PMA), a recently described variant of low-grade astrocytoma is associated with a high rate of recurrence and a propensity for CSF seeding. While cases of PMA have been reported in infants and young children, there has been no report of PMA in patients with neurofibromatosis. The first reportable case of PMA occurring in a child with neurofibromatosis type 1 (NF1) is described. Following presentation with obstructive hydrocephalus, the patient underwent a partial resection of a third ventricular tumor. Histology confirmed the typical features of PMA. The patient demonstrated a partial response to chemotherapy. The authors review the literature on PMA and discuss the specific issues associated with this diagnosis in the context of a child with neurofibromatosis. PMID- 15800887 TI - The Murray Goodman memorial issue. PMID- 15800888 TI - An autonomously folding beta-hairpin derived from the human YAP65 WW domain: attempts to define a minimum ligand-binding motif. AB - WW domains are broadly distributed among natural proteins; these modules play a role in bringing specific proteins together. The ligands recognized by WW domains are short segments rich in proline residues. We have tried to identify the minimum substructure within a WW domain that is required for ligand binding. WW domains typically comprise ca. 40 residues and fold to a three-stranded beta sheet. Structural data for several WW domain/ligand complexes suggest that most or all of the intermolecular contacts involve beta-strands 2 and 3. We have developed a 16-residue peptide that folds to a beta-hairpin conformation that appears to mimic beta-strands 2 and 3 of the human YAP65 WW domain, but this peptide does not bind to known ligands. Thus, the minimum binding domain is larger than the latter two strands of the WW domain beta-sheet. PMID- 15800890 TI - Percutaneous closure of an aortic prosthetic paravalvular leak with an Amplatzer duct occluder. AB - Following aortic valve replacement, up to 5% of patients may develop clinically significant paravalvular leaks. Reoperation is associated with higher mortality and an excess risk of recurrent paravalvular insufficiency. No specifically designed transcatheter device is available for paravalvular leak repair. We describe in a patient with severe aortic prosthetic paravalvular leak the use of an Amplatzer duct occluder device resulting in effective closure and symptomatic improvement. PMID- 15800889 TI - Impact of stents and abciximab on survival from cardiogenic shock treated with percutaneous coronary intervention. AB - This retrospective observational review compares patient characteristics and in hospital and long-term outcomes of cohorts of patients undergoing percutaneous coronary intervention (PCI) for cardiogenic shock complicating acute myocardial infarction (MI) prior to the use of stents (as well as glycoprotein IIb/IIIa inhibitor and dual-antiplatelet therapy) with PCI in the stent era. Cardiogenic shock remains the leading cause of hospital mortality from acute MI. This is a report of consecutive patients with cardiogenic shock complicating acute MI, without mechanical complication, referred for emergency catheterization to a single operator at two consecutive Veterans Affairs medical centers over a 15 year period (1988 to August 2003). PCI was attempted in all 93 cases: 44 consecutive patients in the present era and 49 consecutive patients in the stent era. Patients with comparable extent of coronary disease, more ST elevation myocardial infarction, multiple areas of infarction, and greater comorbidity underwent PCI in the stent era. Nevertheless, PCI in the stent era was associated with higher rates of acute success and improved in-hospital survival. Kaplan Meier curves and log-rank testing showed highly significant improvement in overall survival (P < 0.0001). Logistic regression of in-hospital survival demonstrated that stent use (colinear with glycoprotein IIb/IIIa use and dual antiplatelet therapy) was significantly associated with survival in a model adjusting for extent of coronary disease and comorbidities (P = 0.007). Stents and abciximab have been associated with improved acute angiographic and procedural success of PCI for cardiogenic shock, leading to improved survival. PMID- 15800891 TI - Rapid prototyping to create vascular replicas from CT scan data: making tools to teach, rehearse, and choose treatment strategies. AB - Our goal was to develop and prove the accuracy of a system that would allow us to re-create live patient arterial pathology. Anatomically accurate replicas of blood vessels could allow physicians to teach and practice dangerous interventional techniques and might also be used to gather basic physiologic information. The preparation of replicas has, until now, depended on acquisition of fresh cadaver material. Using rapid prototyping, it should be able to replicate vascular pathology in a live patient. We obtained CT angiographic scan data from two patients with known arterial abnormalities. We took such data and, using proprietary software, created a 3D replica using a commercially available rapid prototyping machine. From the prototypes, using a lost wax technique, we created vessel replicas, placed those replicas in the CT scanner, then compared those images with the original scans. Comparison of the images made directly from the patient and from the replica showed that with each step, the relationships were maintained, remaining within 3% of the original, but some smoothing occurred in the final computer manipulation. From routinely obtainable CT angiographic data, it is possible to create accurate replicas of human vascular pathology with the aid of commercially available stereolithography equipment. Visual analysis of the images appeared to be as important as the measurements. With 64 and 128 slice detector scanners becoming available, acquisition times fall enough that we should be able to model rapidly moving structures such as the aortic root. PMID- 15800892 TI - Electrostatic energies and forces computed without explicit interparticle interactions: a linear time complexity formulation. AB - A rapid method for the calculation of the electrostatic energy of a system without a cutoff is described in which the computational time grows linearly with the number of particles or charges. The inverse of the distance is approximated as a polynomial, which is then transformed into a function whose terms involve individual particles, instead of particle pairs, by a partitioning of the double sum. In this way, the electrostatic energy that is determined by the interparticle interactions is obtained without explicit calculation of these interactions. For systems of positive charges positioned on a face-centered cubic lattice, the calculation of the energy by the new method is shown to be faster than the calculation of the exact energy, in many cases by an order of magnitude, and to be accurate to within 1-2%. The application of this method to increase the accuracy of conventional truncation-based calculations in condensed-phase systems is also demonstrated by combining the approximated long-range electrostatic interactions with the exact short-range interactions in a "hybrid" calculation. For a 20-A sphere of water molecules, the forces are shown to be six times as accurate using this hybrid method as those calculated with conventional truncation of the electrostatic energy function at 12 A. This is accomplished with a slight increase in speed, and with a sevenfold increase in speed relative to the exact all-pair calculation. Structures minimized with the hybrid function are shown to be closer to structures minimized with an exact all-pair electrostatic energy function than are those minimized with a conventional 13-A cutoff-based electrostatic energy function. Comparison of the energies and forces calculated with the exact method illustrate that the absolute errors obtained with standard truncation can be very large. The extension of the current method to other pairwise functions as well as to multibody functions, is described. PMID- 15800893 TI - A Phase I clinical trial of ixabepilone (BMS-247550), an epothilone B analog, administered intravenously on a daily schedule for 3 days. AB - BACKGROUND: The epothilones are a novel class of microtubule-stabilizing agents. Ixabepilone (BMS-247550; NSC 710428) is a semisynthetic analog of the natural product epothilone B. The authors conducted a Phase I study by administering ixabepilone to patients as a 1-hour intravenous infusion daily for 3 consecutive days every 21 days. METHODS: Twenty-six patients were enrolled and received ixabepilone at a starting dose of 8 or 10 mg/m(2) per day for 3 consecutive days. RESULTS: One hundred and nineteen cycles were administered to 26 patients. The maximum-tolerated dose was 8 mg/m(2) per day of ixabepilone administered as a 1 hour intravenous infusion daily for 3 consecutive days every 21 days. The dose limiting toxicity (DLT) was neutropenia. Other nonhematologic Grade 3 toxicities included fatigue (3 cycles), hyponatremia (1 cycle), anorexia (1 cycle), ileus (1 cycle), stomatitis (1 cycle), and emesis (1 cycle). Prolonged disease stabilization was observed in patients with mesothelioma, ovarian carcinoma, and renal cell carcinoma. CONCLUSIONS: The recommended Phase II dose of ixabepilone on the daily schedule for 3 days was 8-10 mg/m(2) per day. Neutropenia was the DLT. Peripheral neuropathy was mild, even after multiple cycles of therapy, and was not dose limiting. PMID- 15800894 TI - Screening women at high risk for breast cancer with mammography and magnetic resonance imaging. AB - BACKGROUND: The authors compared the performance of screening mammography versus magnetic resonance imaging (MRI) in women at genetically high risk for breast cancer. METHODS: The authors conducted an international prospective study of screening mammography and MRI in asymptomatic, genetically high-risk women age >/= 25 years. Women with a history of breast cancer were eligible for a contralateral screening if they had been diagnosed within 5 years or a bilateral screening if they had been diagnosed > 5 years previously. All examinations (MRI, mammography, and clinical breast examination [CBE]) were performed within 90 days of each other. RESULTS: In total, 390 eligible women were enrolled by 13 sites, and 367 women completed all study examinations. Imaging evaluations recommended 38 biopsies, and 27 biopsies were performed, resulting in 4 cancers diagnosed for an overall 1.1% cancer yield (95% confidence interval [95%CI], 0.3-2.8%). MRI detected all four cancers, whereas mammography detected one cancer. The diagnostic yield of mammography was 0.3% (95%CI, 0.01-1.5%). The yield of cancer by MRI alone was 0.8% (95%CI, - 0.3-2.0%). The biopsy recommendation rates for MRI and mammography were 8.5% (95%CI, 5.8-11.8%) and 2.2% (95%CI, 0.1-4.3%). CONCLUSIONS: Screening MRI in high-risk women was capable of detecting mammographically and clinically occult breast cancer. Screening MRI resulted in 22 of 367 of women (6%) who had negative mammogram and negative CBE examinations undergoing biopsy, resulting in 3 additional cancers detected. MRI also resulted in 19 (5%) false-positive outcomes, which resulted in benign biopsies. PMID- 15800895 TI - Cutting balloon recanalization for femoral artery stenosis in a child: a case report. AB - A chronic femoral artery stenosis associated with an ipsilateral leg length discrepancy was found resistant to high-pressure balloon angioplasty. Improvement in vessel caliber and relief of the stenosis were achieved after a cutting balloon. PMID- 15800896 TI - Folate supplementation in three genetic models: implications for understanding folate-dependent developmental pathways. AB - Supplementation of a pregnant mother's diet with folate has been shown to protect the developing embryo from birth defects in humans as well as rodent animal models. Folate supplementation not only reverses a potential nutritional deficiency; folate effectively prevents defects even when the mother's nutritional status is normal. These findings indicate that folate is able to interact with the molecular pathways that control normal embryonic development. Supplementation studies in animals provide the experimental starting point for the identification of such folate-responsive pathways. This review summarizes the progress to date in understanding the folate response in genetic models of birth defects in the mouse. PMID- 15800897 TI - Purkinje cell dendrites grow in alignment with Bergmann glia. AB - The pattern of growth of Purkinje cell dendrites has been analyzed and related to their interactions with Bergmann glial radial processes. In cerebellar slice cultures from mice expressing green fluorescent protein (GFP) under the glial fibrillary acidic protein (GFAP) promoter, Purkinje cells were transfected and imaged with two-photon microscopy over 2 days. We report that while the Purkinje cell dendritic tree grows, individual dendrites increase or decrease in length. Importantly, we demonstrate that vertical growth of Purkinje cell dendrites occurs primarily in alignment with radial glial processes. These findings suggest that radial glial processes provide a structural substrate for the directional growth of Purkinje cell dendrites, thus influencing the shape of the dendritic tree. PMID- 15800902 TI - Pharmaceutical-mediated inactivation of p53 sensitizes U87MG glioma cells to BCNU and temozolomide. AB - Pifithrin-alpha (PFTalpha) is a small molecule inhibitor of p53. By reversibly blocking apoptosis in response to DNA damage, PFTalpha protects normal cells from lethal doses of gamma-radiation (Komarov et al., Science, 1999;285:1733-7). We examined the effect of PFTalpha on the chemosensitivity of a human cancer in which cell cycle arrest, not apoptosis, is the principle cellular consequence of p53 activation. This was of interest because E6 silencing of p53 sensitizes U87MG astrocytic glioma cells to BCNU and temozolomide (TMZ), cytotoxic drugs that are modestly helpful in the treatment of aggressive astrocytic gliomas. We observed that exposure of U87MG cells to PFTalpha before cytotoxic chemotherapy attenuated p53-mediated induction of p21WAF1 protein levels, sensitizing U87MG cells to BCNU and TMZ. Sensitization of U87MG cells was associated with G1 arrest, delayed entry into S-phase and decreased repair of DNA damage by BCNU. Our findings suggest that in addition to protecting normal cells from the toxic effects of radiation and chemotherapy, small molecule inhibitors of p53, like PFTalpha, might play a role in clinical oncology by sensitizing certain resistant cancers to cytotoxic chemotherapies. PMID- 15800903 TI - Inappropriate tall stature and renal ectopy in a male patient with X-linked congenital adrenal hypoplasia due to a novel missense mutation in the DAX-1 gene. AB - Mutations in DAX-1 gene cause congenital adrenal hypoplasia (AHC). We present a male patient affected by X-linked adrenal hypoplasia congenita due to a novel DAX 1 missense mutation. The mutation V287G affects the C-terminal end of the DAX-1 protein which plays an important role in functioning of the receptor. In addition, our patient presented an inappropriate tall stature and renal ectopy, which have not been described in AHC so far. PMID- 15800904 TI - Oral squamous cell carcinoma cells induce osteoclast differentiation by suppression of osteoprotegerin expression in osteoblasts. AB - The invasion of oral squamous cell carcinoma (SCC) cells into the mandibular bone is a common clinical problem. It has been reported that BHY cells, a human oral SCC cell line, are capable of invading mandibular bone of nude mice. These results led us to examine possible mechanisms of osteoclastogenesis induced by BHY cells using in vitro culture systems. When BHY cells were cocultured with mouse bone marrow cells (BMCs), only few osteoclasts were formed, even though BHY cells express the receptor activator of NF-kappaB ligand (RANKL). However, adding BHY cells to a coculture of mouse primary osteoblasts (POBs) and BMCs markedly induced osteoclastogenesis in the absence of osteotropic factors. Furthermore, another oral SCC cell line, HSC-2, which does not express RANKL, also induced osteoclastogenesis in our cocultures. These effects were significantly, but not completely, inhibited by adding osteoprotegerin (OPG). In addition, we also found that TNFalpha released from these cells partially contributes to osteoclastogenesis via a RANKL-independent mechanism. Adding BHY or HSC-2 cells suppressed mouse OPG mRNA expression and protein production by POBs in cocultures of POBs and human oral SCC cells. This finding is consistent with the result that BHY cells and HSC-2 cells did not enhance osteoclastogenesis in cocultures of BMCs and POBs from OPG-deficient mice. Immunohistochemical analysis showed a reduction of OPG expression in osteolytic lesions as compared to normal lesions from oral SCC patients. Therefore, oral SCC-induced suppression of OPG expression in POBs appears critical for osteoclastogenesis, rather than expression of RANKL in SCC cells. PMID- 15800905 TI - Ethnic group-related differences in CpG hypermethylation of the GSTP1 gene promoter among African-American, Caucasian and Asian patients with prostate cancer. AB - The incidence and mortality of prostate cancer (PC) is approximately 2-fold higher among African-Americans as compared to Caucasians and very low in Asian. We hypothesize that inactivation of GSTP1 genes through CpG methylation plays a role in the pathogenesis of PC, and its ability to serve as a diagnostic marker that differs among ethnic groups. GSTP1 promoter hypermethylation and its correlation with clinico-pathological findings were evaluated in 291 PC (Asian = 170; African-American = 44; Caucasian = 77) and 172 benign prostate hypertrophy samples (BPH) (Asian = 96; African-American = 38; Caucasian = 38) using methylation-specific PCR. In PC cells, 5-aza-dC treatment increased expression of GSTP1 mRNA transcripts. The methylation of all CpG sites was found in 191 of 291 PC (65.6%), but only in 34 of 139 BPH (24.5%). The GSTP1 hypermethylation was significantly higher in PC as compared to BPH in each ethnic group (p < 0.0001). Logistic regression analysis (PC vs. BPH) showed that African-Americans had a higher hazard ratio (HR) (13.361) compared to Caucasians (3.829) and Asian (8.603). Chi-square analysis showed correlation of GSTP1 hypermethylation with pathological findings (pT categories and higher Gleason sum) in Asian PC (p < 0.0001) but not in African-Americans and Caucasian PC. Our results suggest that GSTP1 hypermethylation is a sensitive biomarker in African-Americans as compared to that in Caucasians or Asian, and that it strongly influences tumor progression in Asian PC. Ours is the first study investigating GSTP1 methylation differences in PC among African-American, Caucasian and Asian. PMID- 15800906 TI - Autosomal dominant microtia and ocular coloboma: new syndrome or an extension of the oculo-auriculo-vertebral spectrum? AB - The oculo-auriculo-vertebral (OAV) spectrum is an etiologically heterogeneous condition classically consisting of microtia, hemifacial microsomia, epibulbar dermoids, and vertebral anomalies. Other eye findings described in OAV include upper eyelid colobomas, ptosis, and varying degrees of microphthalmia or even anophthalmia. Iris and/or retinal colobomas have rarely been reported. We describe two familial cases of apparent OAV with ocular colobomas. We postulate that iris and/or retinal colobomas associated with OAV may represent a subgroup within the OAV spectrum with autosomal dominant inheritance, as in the families described herein. Since microtia can result from aberrant migration of neural crest cells into the first and second branchial arches during early embryonic development, and concomitant deficient neural crest migration into the developing eye can lead to ocular coloboma and or iris heterochromia, it may be that the altered gene or genes in our familial cases are involved with regulation of neural crest development. PMID- 15800907 TI - Mental retardation in a boy with anterior cervical hypertrichosis. AB - Anterior cervical hypertrichosis (ACH) is a rare form of localized hypertrichosis with 15 previously reported cases. ACH has been considered to be a dominant phenotype, either X-linked or autosomal [OMIM 600457]. ACH was associated with hereditary motor and sensory neuropathy (HMSN) in one family, in which the proband also exhibited severe chorioretinal degeneration and optic atrophy, probably as a different entity [OMIM 239840]. A Mexican boy with congenital ACH associated with moderate mental retardation, abnormal EEG, mild microcephaly, hypertrichosis on the back, and hallux valgus is presented here. An equal sex ratio found in 16 reported cases as well as the suggestion of a paternal age effect in one report appear most consistent with an autosomal dominant mode of inheritance for this trait. It remains unclear if isolated ACH, ACH-HMSN, or other associated findings reported in patients with ACH, including unusual features found in our case, are part of ACH or fortuitous associations, due to the small number of affected patients and different ascertainment biases present in previous reports. PMID- 15800908 TI - Lacking immunocytological GD2 expression in neuroblastoma: report of 3 cases. AB - Immunocytological bone marrow assessment for contamination with neuroblastoma cells is based on their characteristic GD2 surface staining. Neuroblastoma without GD2 expression have been rarely and only after antibody therapy reported. Conventional cytology was performed using Pappenheim staining. For immunocytology, the APAAP method was utilized with the 14G2a anti-GD2 mouse monoclonal antibody. 7 x 10(5) cells on cytospin preparations were investigated. In 2003, 288 bone marrow samples from 191 neuroblastoma patients were investigated by cytology and immunocytology. Three cases demonstrated GD2 negativity on cytologically unambiguous neuroblastoma cells. Two female cases (94 and 37 months of age) with stage 4 neuroblastoma had GD2 expressing neuroblastoma cells in bone marrow at diagnosis. At 2nd relapse 25 and 23 months after diagnosis and 8 months and 12 months after anti-GD2 antibody treatment (ch14.18), the bone marrow infiltrating neuroblastoma cells lacked GD2 staining. The third patient, a 63-month-old girl with bone marrow replacement by neuroblastoma cells showed at diagnosis a mixture of GD2-unstained tumor clumps and very weakly stained neuroblastoma cells. Neuroblastoma cells may lack GD2 expression at diagnosis and at recurrence. This observation has diagnostic and therapeutic implications. PMID- 15800909 TI - Progressive external ophthalmoplegia: a new family with tremor and peripheral neuropathy. PMID- 15800910 TI - Metastatic malignant melanoma presenting as pancytopenia in a three-year-old boy. AB - Malignant melanoma is rare in childhood and has never been reported to cause pancytopenia due to bone marrow metastases in a child. We report a 3-year-old boy with a large congenital melanocytic nevus who presented with bone pain and pancytopenia due to diffuse bone and bone marrow infiltration with metastatic melanoma without an identifiable primary site. Despite treatment with imatinib mesylate there was no response and the patient died with progressive disease. This case illustrates an unusual presentation of bone marrow failure secondary to malignant melanoma in a young child with symptomatic metastatic marrow infiltration, a rarely reported site of melanoma involvement in adults or children. PMID- 15800911 TI - Activation of human cancer/testis antigen gene, XAGE-1, in tumor cells is correlated with CpG island hypomethylation. AB - Expression of the XAGE-1 antigen is restricted to germ cells of the testis and a variety of neoplastic tissues. To date, the molecular mechanism for regulating expression of this cancer/testis antigen gene has been unknown. To evaluate methylation as a potential mechanism for regulating expression of this gene, we first correlated gene methylation status (measured by sequencing of bisulfide modified DNA and COBRA) to expression of XAGE-1 mRNA in normal and cancerous cells. This analysis revealed dense methylation of the CpG island in the XAGE-1 gene promoter for the normal and cancerous cells that do not express this gene but loss of this methylation in normal testis, cancer cell lines and the primary gastric cancers where the gene is highly expressed. Further supporting the role of methylation in regulating expression of XAGE-1 were observations that treatment of 2 heavily methylated cell lines, SNU620 and HT29, with 5'-aza deoxycytidine resulted in demethylation of XAGE-1 promoter and corresponding expression of this gene. Finally, we cloned various segments of the CpG-rich XAGE 1 gene promoter linked to a luciferase reporter construct and transiently transfected this construct into HCT116 cells. These experiments confirmed transcriptional regulatory activity for the promoter region that incorporates the CpG island and demonstrated that in vitro methylation of this island results in loss of promoter activity. Collectively, these studies indicate that XAGE-1 expression in normal and cancerous tissues is regulated by methylation of the CpG island in the gene promoter. PMID- 15800912 TI - Recombinant adeno-associated virus-mediated TRAIL gene therapy suppresses liver metastatic tumors. AB - To evaluate the tumoricidal activity of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on disseminated liver metastatic tumors, we constructed a recombinant adeno-associated virus (rAAV) expressing the extracellular domain (95-281aa) of human TRAIL (TRAIL(95-281), and the recombinant virus was designated as rAAV-TRAIL) using the 3-plasmid, helper-virus free, packaging system. Transduction of mouse lymphoma EL-4 cells and Jurkat T cells lead to the expression of TRAIL(95-281) protein in both virus-transduced cells and the culture media, along with apoptosis of these cells in vitro. The therapeutic potential of rAAV-TRAIL was then evaluated in an orthotopic transplanted mouse model mimicking liver cancer metastasis, which was established by injection of EL-4 cells into the liver of C57BL/6 mice via the hepatic portal veins. Subsequent intraportal vein injection of rAAV-TRAIL, not the control virus, into the liver of these mice resulted in significant suppression of tumor growth and prolonged survival, while normal hepatocyte toxicity is undetectable. Histological and biochemical analysis in tumor tissue and serum confirmed that TRAIL(95-281) was stably expressed in relatively high level in hepatocytes and was secreted into the serum in active trimeric form. Futhermore, the mechanism for rAAV-TRAIL to inhibit tumor growth was by inducing apoptosis of the tumor cells metastasizing to the livers. These results strongly suggest that the rAAV TRAIL-mediated gene delivery could be a promising approach for the treatment of liver metastasis cancer. PMID- 15800913 TI - Engineered vascular-targeting antibody-interferon-gamma fusion protein for cancer therapy. AB - A number of cytokines are either approved drugs or are in advanced clinical trials, yet these biopharmaceuticals do not typically localize efficiently in solid tumors and manifest their therapeutic potential at the expense of severe side effects. The targeted delivery of cytokines to solid tumors is a promising avenue for increasing the therapeutic index of these biopharmaceuticals. We engineered a fusion protein between scFv(L19), a human antibody fragment specific to the EDB domain of fibronectin, and a cysteine-free mutant of murine interferon gamma. The resulting fusion protein was capable of targeting new blood vessels in solid tumors, and the targeting efficiency was strikingly increased in tumor bearing knockout mice lacking the interferon-gamma receptor. ScFv(L19)-interferon gamma displayed a strong antitumor effect in both subcutaneous and metastatic murine F9 teratocarcinomas, but was not efficacious as single agent when used to treat C51 and CT26 tumors. The potency of this fusion protein could be substantially enhanced by combination with doxorubicin and other immunocytokines. These findings are of clinical relevance, as the EDB domain is a marker of angiogenesis, with identical sequence in mouse and man, which is abundantly expressed in a variety of aggressive solid tumors but is undetectable in most normal tissues. PMID- 15800914 TI - Dendritic cells delivered inside human carcinomas are sequestered by interleukin 8. AB - In the course of a clinical trial consisting of intratumoral injections of dendritic cells (DCs) transfected to produce interleukin-12, the use of (111)In labeled tracing doses of DCs showed that most DCs remained inside tumor tissue, instead of migrating out. In search for factors that could explain this retention, it was found that tumors from patients suffering hepatocellular carcinoma, colorectal or pancreatic cancer were producing IL-8 and that this chemokine attracted monocyte-derived dendritic cells that uniformly express both IL-8 receptors CXCR1 and CXCR2. Accordingly, neutralizing antihuman IL-8 monoclonal antibodies blocked the chemotactic attraction of DCs by recombinant IL 8, as well as by the serum of the patients or culture supernatants of human colorectal carcinomas. In addition, tissue culture supernatants of colon carcinoma cells inhibited DC migration induced by MIP-3beta in an IL-8-dependent fashion. IL-8 production in malignant tissue and the responsiveness of DCs to IL 8 are a likely explanation of the clinical images, which suggest retention of DCs inside human malignant lesions. Impairment of DC migration toward lymphoid tissue could be involved in cancer immune evasion. PMID- 15800915 TI - Melatonin potentiates flavone-induced apoptosis in human colon cancer cells by increasing the level of glycolytic end products. AB - Melatonin is a natural compound synthesized by a variety of organs. It has been described to possess cell protecting activity in normal cells but was shown to induce apoptotic cell death in cancer cells. We determined to which extent and based on which molecular mechanisms melatonin is able to cause apoptosis in HT-29 human colon cancer cells. Induction of apoptosis was assessed by caspase-3-like activity, nuclear fragmentation and chromatin condensation. Melatonin, when given alone at a concentration of 1 mM, did not affect any of the apoptosis markers. It potentiated apoptosis induced by the flavonoid flavone significantly. Whereas flavone alone at a concentration of 150 microM led to a 8-fold increase in caspase-3-like activity associated with around 40% of cells displaying DNA fragmentation, a combination of flavone and melatonin increased caspase-3-like activity 30-fold and 80% of cells exhibited fragmentation of DNA when compared to untreated controls. Melatonin caused an increase in cytosolic lactate levels that most likely allows the flavone-induced activation of the mitochondrial pyruvate/lactate importer to deliver more substrates to mitochondrial respiration. The subsequent increased production of mitochondrial O2-* in the presence of flavone was further increased by melatonin. Scavenging mitochondrial O2-* by benzoquinone or blocking the lactate/pyruvate transporter by 5-nitro-2-(3 phenylpropylamino) benzoate inhibited mitochondrial O2-* -generation and apoptosis execution mediated by flavone and melatonin. Our study provides evidence that melatonin potentiates flavone-induced apoptosis in HT-29 human colon cancer cells by enhancing the level of oxidizable substrates that can be transported into mitochondria in the presence of flavone. PMID- 15800916 TI - Overexpression of S100A2 protein as a prognostic marker for patients with stage I non small cell lung cancer. AB - S100A2, a calcium-binding protein, recently became of major interest because of its differential expression during transformation and metastasis in various tumors. The purpose of this study was to investigate the prognostic significance of S100A2 expression in the early-stage non small lung cancer (NSCLC). Immunohistochemical analysis to determine the percentage of cells staining positive for S100A2 was performed on 11 NSCLC tissue microarray slides containing samples from 113 patients with pathologic stage I NSCLC who had undergone curative surgery. S100A2 was expressed in samples from 79 patients (69.9%). Kaplan-Meier analysis showed that patients whose tumors had positive S100A2 expression had a significantly lower overall survival and disease-specific survival rate at 5 years after surgery than did patients with negative S100A2 expression (p < 0.001 and p < 0.001, respectively). Age at diagnosis, histologic type of cancer, degree of differentiation and smoking history did not have a statistically significant effect on survival. Multivariate analysis confirmed that S100A2 expression is a better predictor for disease-specific survival than were other clinical and histologic variables tested. Our results suggested that the expression of the S100A2 protein in stage I NSCLC indicates poor prognosis and may be used to identify patients with early-stage NSCLC who might benefit from adjuvant treatment. PMID- 15800917 TI - Is there an association between maternal weight and the risk of testicular cancer? An epidemiologic study of Norwegian data with emphasis on World War II. AB - Since registration started in the 1950s, the incidence of testicular cancer (TC) in the Western world has increased, which is also the case in Norway. Men born in Norway during World War II (WWII), however, have a lower TC incidence than men born in the years before or after WWII. Increased fetal exposure to estrogen during the first trimester of pregnancy has been proposed as a risk factor for the development of TC later in life. Increased maternal weight is associated with higher insulin levels, leading to lower sex hormone-binding globulin levels and thereby increased levels of bioavailable estrogens for transplacental transfer from mother to fetus. The aim of the present study was therefore to examine whether there was an association between maternal weight and the incidence of TC among those who were born in a time period where the nutritional conditions changed, i.e., around the time of WWII. We compared data for a random sample of women giving birth in Oslo, Norway, in the years 1931 to 1955 with the TC incidence among men born in the whole country in the same time period. Maternal weight at delivery was used as a proxy for first-trimester weight. We found a correlation (Spearman's rho = 1.00, p < 0.01; Pearson's r = 0.95, p = 0.02) between the TC incidence and maternal weight adjusted for birth weight and maternal age. Although one cannot draw firm conclusions from ecologic correlations, these findings suggest that the increase in TC incidence over the past decades could be at least partly attributed to the increased maternal body weight observed in most populations in the relevant time period since TC is thought to be associated with in utero conditions. PMID- 15800918 TI - Mechanisms of tumor vascular shutdown induced by 5,6-dimethylxanthenone-4-acetic acid (DMXAA): Increased tumor vascular permeability. AB - The novel vascular targeting agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA) has completed phase 1 clinical trial and has shown tumor antivascular activity in both mice and humans. We have investigated its ability to change tumor vascular permeability, relating it to tumor vascular perfusion and other responses. The murine colon 38 adenocarcinoma was grown in C57Bl wild-type mice and mice lacking expression of either tumor necrosis factor receptor-1 (TNFR1(-/-)) or TNF (TNF-/ ). Tumor vascular permeability, as measured by extravasation of albumin-Evans Blue complexes 4 hr after DMXAA treatment, was significantly increased in tumor tissue in C57Bl, TNFR1-/- and TNF-/- mice but not in normal (skin) tissue. Significant linear relationships were found between increased tumor vascular permeability, decreased functioning tumor blood vessels (measured by Hoechst 33342 staining at 4 hr), increased plasma 5-hydroxyindole-3-acetic acid concentrations (as a measure of serotonin release by platelets) and the degree of induced tumor hemorrhagic necrosis. The results support the hypothesis that DMXAA increases tumor vascular permeability both directly and through the induction of other vasoactive mediators, including TNF. DMXAA might be useful clinically to potentiate the vascular permeability of other anticancer modalities such as cytotoxic drugs, antibodies, drug conjugates and gene therapy. PMID- 15800919 TI - Five-year follow-up of health-related quality of life after primary treatment of localized prostate cancer. AB - Although with earlier detection of prostate cancer more men face the long-term consequences of primary treatment, studies on the impact of treatment on long term health-related quality of life (HRQoL) are scarce. We followed 314 men with newly diagnosed localized prostate cancer from 1 month before until 5 years after radical prostatectomy (n = 127) or external beam radiotherapy (n = 187; median follow-up = 52 months). Questionnaires addressing disease-specific (UCLA PCI) and generic (SF-36, EQ-5D) HRQoL were sent 1 month before and 6, 12 and 52 months after treatment. Repeated-measures modeling was used to study HRQoL over time. Regular urinary leakage was reported by 12% of prostatectomy patients before treatment and by 31% at the 52-month assessment. Erectile dysfunction before treatment was reported by 31% of prostatectomy patients and by 40% of radiotherapy patients; at the 52-month assessment, these percentages were 88% and 64%, respectively. Erectile dysfunction present at 1 year posttreatment can be considered permanent. Prostatectomy patients reported better generic functioning both before and after treatment than radiotherapy patients, who were on average 5.9 years older and had more comorbid conditions. General physical functioning of prostatectomy patients slightly improved over time, but declined in radiotherapy patients. The relation between age and physical scores was found to be nonlinear. The long-term physical decline in radiotherapy patients partly resulted from aging and its nonlinear impact on health, although treatment effects cannot be excluded. Scores of both patient groups remained above those of norm populations. Innovative graphs describing disease-specific and generic functions after treatment can help patients and physicians in their treatment choices. PMID- 15800920 TI - Expression patterns of mitotic and meiotic cell cycle regulators in testicular cancer and development. AB - Mitotic and meiotic cell cycle regulation is essential for normal development and tumor prevention. The underlying molecular mechanisms are not completely characterized. The aim of our analysis was to derive a global expression map of cell cycle regulators in mitosis and meiosis. First, the expression of cyclins, CDKs and CDK inhibitors was determined during postnatal testis maturation in mice using microarrays and quantitative RT-PCR. The abundance of cyclins A1, B2, K, M4, CDK2, all CDKLs, CDKN2c, CDKN2d and INCA1 increased during testis maturation. In contrast, cyclins A2, B1, D2, G1, G2, CDK1, CDK4 and CDK2AP1 showed a maturation-associated decrease. Gene expression profiles of isolated germ cells and testicular somatic cells confirmed these results. Second, we determined cyclin expression patterns in human normal and malignant testis samples (n = 36) modeling the reciprocal difference between meiosis and mitosis. Testicular tumors strictly expressed cell cycle regulators identified in mitotically dividing germ cells. Expression of several transcripts was histology-specific in testicular tumors, providing novel molecular markers and potential therapeutic targets. Taken together, our data provide a comprehensive expression map of cell cycle regulators at the switch between mitosis and meiosis in testis development and in cancerogenesis. PMID- 15800921 TI - Sequential, randomized trial of a low-fat, high-fiber diet and soy supplementation: effects on circulating IGF-I and its binding proteins in premenopausal women. AB - Despite evidence supporting the involvement of the IGF system in the development of breast and other cancers, the major determinants of interindividual variability in circulatory IGF-I levels are not well understood. Previous research has pointed to important genetic influences as well as dietary effects through marked calorie or protein restriction. We conducted a randomized trial to determine the effects of 2 dietary patterns on serum IGF-1, IGFBP1 and IGFBP3 in free-living premenopausal women: phase 1, an isocaloric low-fat, high-fiber (LFHF) vs. usual diet, and phase 2, a soy supplement either with or without isoflavones (soy+IF vs. soy-IF). Participants completed 12 menstrual cycles on phase 1 and then were randomly assigned to a soy supplement for 3 cycles while maintaining the phase 1 diet. Before and after each phase, 154 women provided serum. We found no difference in the change in IGF-I, BP1 or BP3 in the LFHF group compared to the usual diet group. In phase 2, there were no differences in any IGF protein between the soy+IF and the soy-IF groups or any evidence of interaction between isoflavone exposure and the background diet. However, there was a small but statistically significant decrease (2.3%) in BP3 and an increase in the IGF-I:BP3 molar ratio among all 153 subjects following either soy supplement. These changes were correlated with changes in intake of calcium, total vegetable protein and soy. The results are compatible with previous data suggesting that increases in dietary calcium, protein and soy, in particular, could increase circulating levels of bioavailable IGF-I. PMID- 15800922 TI - Antioxidant vitamin and mineral supplementation and prostate cancer prevention in the SU.VI.MAX trial. AB - Randomized trials have shown, unexpectedly, that supplementation with selenium or vitamin E is associated with a reduction of prostate cancer risk. We assess whether a supplementation with low doses of antioxidant vitamins and minerals could reduce the occurrence of prostate cancer and influence biochemical markers. The SU.VI.MAX trial comprised 5,141 men randomized to take either a placebo or a supplementation with nutritional doses of vitamin C, vitamin E, beta-carotene, selenium and zinc daily for 8 years. Biochemical markers of prostate cancer risk such as prostate-specific antigen (PSA) and insulin-like growth factors (IGFs) were measured on plasma samples collected at enrollment and at the end of follow up from 3,616 men. Cox regression models were used to estimate the hazard ratio and related 95% confidence interval of prostate cancer associated with the supplementation and to examine whether the effect differed among predetermined susceptible subgroups. During the follow-up, 103 cases of prostate cancer were diagnosed. Overall, there was a moderate nonsignificant reduction in prostate cancer rate associated with the supplementation (hazard ratio = 0.88; 95% CI = 0.60-1.29). However, the effect differed significantly between men with normal baseline PSA (< 3 microg/L) and those with elevated PSA (p = 0.009). Among men with normal PSA, there was a marked statistically significant reduction in the rate of prostate cancer for men receiving the supplements (hazard ratio = 0.52; 95% CI = 0.29-0.92). In men with elevated PSA at baseline, the supplementation was associated with an increased incidence of prostate cancer of borderline statistical significance (hazard ratio = 1.54; 95% CI = 0.87-2.72). The supplementation had no effect on PSA or IGF levels. Our findings support the hypothesis that chemoprevention of prostate cancer can be achieved with nutritional doses of antioxidant vitamins and minerals. PMID- 15800923 TI - CDC91L1 (PIG-U) mRNA expression in urothelial cell carcinomas. AB - CDC91L1 (PIG-U) was recently discovered as a new oncogene in human bladder cancer and showed mRNA overexpression in 36% of primary bladder tumor tissues compared to normal urothelium. We further investigated CDC91L1 mRNA expression in 8 bladder cancer cell lines, 14 normal bladder tissues and 42 urothelial cell carcinomas by real-time quantitative PCR. The prognostic value of CDC91L1 mRNA expression was also investigated. Surprisingly, only one (2.4%) tumor tissue showed overexpression compared to normal urothelium. No significant relationship of CDC91L1 mRNA expression with increasing pathologic stage (p = 0.962) or grade (p = 0.557) was observed. Median normalized CDC91L1 mRNA expression values were 0.19 for superficial tumors (n = 21) and 0.18 for invasive tumors (n = 21). Grade I, grade II and grade III tumors had median normalized expression values of 0.26, 0.18 and 0.33, respectively. CDC91L1 mRNA expression level was not indicative of early tumor recurrence (log rank p = 0.1629), tumor progression (log rank p = 0.9307) or overall and disease-specific survival (log rank p = 0.9193 and 0.4710, respectively). Our results suggest, in contrast to those of Guo et al. (Nat Med 2004;10:374-81), that the oncogene CDC91L1 is not overexpressed at the mRNA level in urothelial cell carcinomas and cannot be used to predict the course of the disease. PMID- 15800924 TI - CYP19 (aromatase) haplotypes and endometrial cancer risk. AB - Endogenous estrogen exposure is an important determinant of endometrial cancer risk. Aromatase, encoded by CYP19, catalyzes the aromatization of androstenedione and testosterone to estrone and estradiol, respectively. Several common genetic polymorphisms in CYP19 have been identified, including a TCT insertion/deletion and a (TTTA)(n) repeat polymorphism in intron IV as well as a 3'UTR C/T polymorphism. We evaluated these 3 polymorphisms plus an additional 9 noncoding polymorphisms as individual genotypes and predicted haplotypes as risk factors for endometrial cancer using a nested case-control study design. Invasive endometrial cancer cases (n = 222) and matched controls (n = 666) were identified among participants in the Nurses' Health Study who had provided a blood sample in 1989-1990 (n = 32,826). We estimated haplotypes from unphased genotype data spanning > 123 kb of CYP19. Six haplotypes constructed from 10 SNPs were estimated with a frequency > or = 5%. The highest prevalence haplotype (33% among cases, 28% among controls) was significantly associated with endometrial cancer risk (p = 0.03). Loci with variant alleles that comprise the risk haplotype were independently associated with endometrial cancer, with relative risk estimates ranging from 1.68 (95% CI 1.13-2.48) to 2.07 (95% CI 1.33-3.23), comparing variant allele carriers to wild-type homozygotes. We observed significant interactions between menopausal status and 2 of the high-risk loci (p = 0.03 and p < 0.01), with > 2-fold increased risk for variant allele carriers who were postmenopausal but no association between genotype and endometrial cancer among premenopausal women. We evaluated associations between CYP19 haplotypes and plasma steroid hormone levels. The haplotype associated with endometrial cancer risk is also significantly associated with the ratios of estrone to androstenedione and estradiol to testosterone, the products and substrates of the enzyme aromatase, encoded by CYP19. Our data suggest that there is a high frequency CYP19 haplotype related to higher estrogen to androgen ratios and increased risk of endometrial cancer and that this association may primarily pertain to postmenopausal women. PMID- 15800925 TI - Signal transduction pathways regulating cyclooxygenase-2 in lipopolysaccharide activated primary rat microglia. AB - Microglia are the major cell type involved in neuroinflammatory events in brain diseases such as encephalitis, stroke, and neurodegenerative disorders, and contribute significantly to the release of prostaglandins (PGs) during neuronal insults. In this report, we studied the immediate-early intracellular signalling pathways in microglia, following bacterial lipopolysaccharide (LPS) stimulation, leading to the synthesis and release of PGE2. Here we show that LPS induces cyclooxygenase (COX) 2 by activating sphingomyelinases leading to the release of ceramides, which in turn, activate the p38 mitogen-activated protein kinases (MAPK), but not the p42/44 MAPK. We further show that exogenously added ceramide analogue (C2-ceramide) also induce PGE2 synthesis through a p38 MAPK-dependent pathway. This potential nature of ceramides in activating microglia suggests that endogenously produced ceramides during neuronal apoptosis in ischemia or neurodegenerative diseases could also contribute to the amplification of neuroinflammatory events. In contrast to protein kinase C (PKC) and phosphocholine-specific phospholipase C (PC-PLC), which transcriptionally regulate LPS-induced COX-2 synthesis, inhibition of phospholipase A2 (PLA2) has no effect on COX-2 transcription, although it inhibits the release of PGE2. Transcriptional regulation of LPS-induced COX-2 by PKC is further proved by the ability of the PKC inhibitor, Go 6976, to inhibit LPS-induced 8-isoprostane synthesis, but not affecting LPS-induced COX-2 activity. Our data with 8 isoprostane also indicates that COX-2 plays a major role in ROS production in LPS activated microglia. This detailed view of the intracellular signaling pathway in microglial activation and COX-2 expression opens a new therapeutic window in the search for new and more effective central anti-inflammatory agents. PMID- 15800926 TI - A paracrine signal mediates the cell transformation response to low dose gamma radiation in JB6 cells. AB - The carcinogenic response to radiation is complex and may involve adaptive cellular responses as well as a bystander effect mediated by paracrine or intercellular signaling activities. Using a newly developed co-culture model we have examined whether low dose gamma radiation induces the transformation of JB6 mouse epidermal cells as well as non-irradiated bystander cells. Cell transformation response is defined as the acquisition of anchorage-independent growth properties and is quantified by counting colonies on soft agar. Exposure of JB6 cells to low dose (2-20 cGy) gamma radiation resulted in an approximate 1.9 +/- 0.1 and 2.8 +/- 0.5-fold increase in cell transformation response when cells were seeded at 1 x 10(4) or 1 x 10(5) cells/dish, relative to respective sham exposed controls. We developed a co-culture model where sham exposed or irradiated JB6 cells were mixed with non-irradiated JB6 cells that had been stably transfected with the enhanced yellow fluorescent protein (EYFP) to enable the distinction of fluorescent bystander-specific colonies. A significant increase in the number of bystander-specific colonies was observed in co-culture with 10 cGy irradiated JB6 cells (224 +/- 9), relative to the number of bystander specific colonies arising in co-culture with sham exposed JB6 cells (55 +/- 16). Our results indicate that low dose radiation induces the transformation of JB6 cells and that a soluble paracrine factor that is secreted by irradiated cells induces the transformation of non-irradiated bystander cells. PMID- 15800927 TI - E1A-F is overexpressed early in human colorectal neoplasia and associated with cyclooxygenase-2 and matrix metalloproteinase-7. AB - Studies suggest the expression of cyclooxygenase-2 (COX-2) and matrilysin (MMP-7) increase in the early stages of colorectal carcinogenesis, however their interaction with other molecular markers is poorly understood. Results from cell line studies and mouse models suggest polyomavirus enhancer activator 3 (PEA3) may play a role in the activation of COX-2 and MMP-7 promoters. However, the role of E1A-F, the human homolog of murine PEA3, in colorectal cancer (CRC) development has not been elucidated. In this study, we used real-time reverse transcription (RT)-polymerase chain reaction (PCR) to measure the levels of E1A F, COX-2, and MMP-7 in matched normal mucosa, adenomas, and/or carcinomas from 128 patients. Our results demonstrate significant overexpression of E1A-F and MMP 7 in adenomas and E1A-F, COX-2, and MMP-7 in carcinomas. In carcinomas, E1A-F expression was significantly associated with both COX-2 and MMP-7 overexpression. These results suggest E1A-F is overexpressed in early stages of human CRC development and may be an important factor in the overexpression of COX-2 and MMP 7. PMID- 15800928 TI - Induction of blood-brain barrier properties in cultured brain capillary endothelial cells: comparison between primary glial cells and C6 cell line. AB - The communication between glial cells and brain capillary endothelial cells is crucial for a well-differentiated blood-brain barrier (BBB). It has been suggested that in vitro primary glial cells (GCs) be replaced by the glial C6 cell line to standardise the model further. This study compares directly the structural and functional differentiation of bovine brain capillary endothelial cells (BBCECs) induced by co-culture with rat primary GCs or C6 cells, for the first time. Trans-endothelial electrical resistance (TEER) measurements showed that under no condition were C6 cells able to reproduce TEER values as high as in the presence of GCs. At the same time, permeability of the BBCECs to both radioactive sucrose and FITC-inulin was 2.5-fold higher when cells were co cultured with C6 than with GCs. Furthermore, immunocytochemistry studies showed different cell morphology and less developed tight junction pattern of BBCECs co cultured with C6 toward GCs. Additionally, studies on P-glycoprotein (P-gp) showed much lower P-gp presence and activity in BBCECs co-cultured with C6 than GCs. Both VEGF mRNA expression and protein content were dramatically increased when compared with GCs, suggesting that VEGF could be one of the factors responsible for higher permeability of BBB. Our results clearly indicate that, in the presence of the glial C6 cell line, BBCECs did not differentiate as well as in the co-culture with primary GCs at both structural and functional levels. PMID- 15800929 TI - Combined subcarcinogenic benzo[a]pyrene and UVA synergistically caused high tumor incidence and mutations in H-ras gene, but not p53, in SKH-1 hairless mouse skin. AB - Combined subcarcinogenic doses of benzo[a]pyrene (BaP) and UVA induced H-ras, but not p53, gene mutations 8 weeks before tumor emergence in SKH-1 mice. Neither UVA (40 kJ/m2) nor BaP (8 nmol) induced any tumors after mice were topically treated 3 times/week for 25 weeks. However, combined BaP-UVA treatment synergistically increased tumor incidence and multiplicity. All tumors induced by BaP-UVA were malignant. The epidermis was collected from mice treated for 2, 6 and 10 weeks. DNA from UVB- (0.3 kJ/m2) or BaP-UVA-(8 nmol and 40 kJ/m2-induced tumors was isolated and screened for H-ras and p53 mutations. Four types of point mutation, GGC-->GAC, GCC, GTC and CGC, occurred in UVB-induced tumors at H-ras codon 13; and one type of point mutation, GGA-->GAA, at codon 12. Treatment with either BaP alone or BaP-UVA for 10 weeks caused GGA-->GAA mutation at codon 12 or GGC-->GAC mutation at codon 13 in nontumor skin, respectively, as well as in tumors induced by BaP-UVA. All of the 10-week samples treated with either BaP or BaP-UVA showed detectable mutations at codons 12 and 13, but the genetic load was significantly higher in BaP-UVA-treated mice than in those exposed only to BaP. UVA alone induced mutations at codon 12 in only one-third of samples. G-->A mutations induced by BaP or BaP-UVA at position 38 of codon 13 have not been reported previously. C-->T transitions were detected in p53 hot spots of exon 8 in 2 of 19 BaP-UVA-induced tumors but were not found in nontumor skin. PMID- 15800930 TI - Stem/progenitor cell proliferation factors FGF-2, IGF-1, and VEGF exhibit early decline during the course of aging in the hippocampus: role of astrocytes. AB - Dentate neurogenesis, important for learning and memory, declines dramatically by middle age. Although studies have shown that this age-related decrease can be reversed to some extent by exogenous applications of mitogenic factors, it is unclear whether one or more of these factors exhibits decline by middle age. We hypothesize that multiple stem/progenitor cell proliferation factors exhibit early decline during the course of aging in the hippocampus, and some of these declines are linked to age-related alterations in hippocampal astrocytes. We measured the concentrations of fibroblast growth factor-2 (FGF-2), insulin-like growth factor-1 (IGF-1), and vascular endothelial growth factor (VEGF) in the hippocampus of young, middle-aged, and aged F344 rats, using enzyme-linked immunosorbent assay (ELISA). In addition, we quantified the total number of FGF-2 immunopositive (FGF-2+) and glial fibrillary acidic protein immunopositive (GFAP+) cells in the dentate gyrus and the entire hippocampus. Our results provide new evidence that the concentrations of FGF-2, IGF-1, and VEGF decline considerably by middle age but remain steady between middle age and old age. Further, decreased concentrations of FGF-2 during aging are associated with decreased numbers of FGF-2+ astrocytes. Quantification of GFAP+ cells, and GFAP and FGF-2 dual immunostaining analyses, reveal that aging does not decrease the total number of astrocytes but fractions of astrocytes that express FGF-2 decline considerably by middle age. Thus, dramatically decreased dentate neurogenesis by middle age is likely linked to reduced concentrations of FGF-2, IGF-1, and VEGF in the hippocampus, as each of these factors can individually influence the proliferation of stem/progenitor cells in the dentate gyrus. Additionally, the results demonstrate that decreased FGF-2 concentration during aging is a consequence of age-related impairment in FGF-2 synthesis by astrocytes. PMID- 15800931 TI - Phenotypic persistence after reoxygenation of hypoxic neuroblastoma cells. AB - Fast-growing solid tumors are usually insufficiently vascularized, leading to areas with necrosis and/or poorly oxygenated cells. Tumor cells adapt to acute hypoxic stress. Central to this adaptation are the hypoxia-inducible transcription factors (HIFs), which are degraded at normoxic but become stabilized at hypoxic conditions. Hypoxic (1% O2) neuroblastoma cells downregulate sympathetic nervous system marker genes, whereas neural crest cell markers are upregulated, suggesting that hypoxic tumor cells adopt a less mature phenotype, which in the clinical setting would translate to more aggressive tumors with increased metastatic potential. Here, we compared gene expression patterns in neuroblastoma cells grown at 1%, 5% (a physiologic oxygen level) and 21% O2. At 5% O2, cells developed a weak hypoxic phenotype and HIF-2 alpha, but not HIF-1 alpha, was acutely stabilized. At 1% O2, HIF-2 alpha protein remained present in long-term cultures, while HIF-1 alpha was present only transiently. The stability of the hypoxia-induced dedifferentiated phenotype in cells acutely reoxygenated at either 21% or 5% O2 persisted for at least 24 hr. Genes associated with a differentiated state, like NPY, ChrA and ChrB, were still downregulated and hypoxia-induced genes, like TH and Id2, remained upregulated. Thus, if these culture conditions are viewed as models for acute reoxygenation of metastasizing hypoxic tumor cells, our data suggest that an aggressive hypoxic phenotype persists for 24 hr or more, which might be long enough for the cells to be able to home to secondary sites, in part as a consequence of their immature hypoxic characteristics. PMID- 15800932 TI - In vivo and in vitro antitumor activity of butyroyloxymethyl-diethyl phosphate (AN-7), a histone deacetylase inhibitor, in human prostate cancer. AB - AN-7, a prodrug of butyric acid, induced histone hyperacetylation and differentiation and inhibited proliferation of human prostate 22Rv1 cancer cells in vitro and in vivo. In nude mice implanted with these cells, 50 mg/kg AN-7 given orally thrice a week led to inhibition of tumor growth and metastasis, tumor regression in >25% of animals and increased survival. Median time to the experimental end point (tumor volume 2 cm3 or death) in the untreated was 52 days, and average tumor volume was 0.8 +/- 0.18 cm3. At the same time, 94.4% of AN-7-treated mice survived and had average tumor volumes of 0.37 +/- 0.1 cm3. PSA expression was a useful marker for 22Rv1 lung metastasis detection. Sizeable metastases positively stained for PSA and limited air gaps were found in lungs of untreated mice. In animals treated with AN-7, lung morphology appeared normal. Primary tumors of treated animals were highly positive for PSA and had an elevated level of p21 and the proapoptotic protein Bax. Sections taken from AN-7 treated animals, examined under an electron microscope, exhibited condensed chromatin and apoptotic bodies. PSA serum levels were higher in untreated compared to treated animals and correlated with tumor volume. Since prolonged oral administration with 50 mg/kg or a single oral dose of 1.2 g/kg AN-7 did not cause adverse effects and the former exhibited significant anticancer activity, AN-7 is likely to display a high therapeutic index and may be beneficial for prostate cancer patients. PMID- 15800933 TI - Presence and specificity of tumor associated lymphocytes from ascites fluid in prostate cancer. AB - BACKGROUND: In the present study, we had the rare opportunity to study immunological responses of TAL from ascites fluid in a patient with hormone refractory prostate cancer. METHODS: We evaluated tumor antigen-specific T-cell responses, induced by either prostate specific antigen (PSA) pulsed dendritic cells (DCs) or PSA peptides, in TAL and peripheral blood lymphocytes. RESULTS: DC stimulation with PSA protein induced recognition of naturally processed PSA epitopes by both blood and ascites T cells. In contrast, only ascites T cells recognized the PSA-3 peptide, after stimulation with PSA-pulsed DCs or peptides. Finally, although IFNgamma secreting T cells were detectable in both blood and ascites by ELISPOT, multiplex cytokine assay detected the presence of predominantly Th2 cytokines. CONCLUSIONS: Although tumor antigen-specific TAL were detected in ascites fluid, these cells were producing immunosuppressive cytokines which may contribute to tumor escape from recognition and/or destruction by the immune system. PMID- 15800934 TI - IGF-I and IGFBP-3 polymorphisms and risk of prostate cancer. AB - BACKGROUND: Insulin-like growth factor-I (IGF-I) is a potent mitogen for both normal and malignant prostate epithelial cells. The majority of circulating IGF-I is bound in a complex with IGF binding protein-3 (IGFBP-3), which in turn limits IGF-I bioavailability. Multiple studies suggest that higher IGF-I and/or lower IGFBP-3 serum levels are positively associated with prostate cancer risk. Several polymorphisms within the IGF-I and IGFBP-3 coding regions have been associated with increased serum protein levels. METHODS: To ascertain the potential relationship between serum levels and polymorphism, and prostate cancer risk, we investigated the role of two polymorphisms the IGF-I cytosine-adenosine (CA) repeat and the IGFBP-3 Ala32Gly, and prostate cancer in a population-based, case control, study of middle-aged men. RESULTS: We found no significant association between the IGFBP-3 Ala32Gly polymorphism and prostate cancer risk, even though the presence of at least one Gly allele did correlate with increased serum levels of IGFBP-3. For IGF-I, more controls (42%) than cases (38%) were homozygous for 19-CA-repeats (odds ratio, OR = 0.85; 95% confidence interval (CI) = 0.66-1.09). After stratifying by disease characteristics, 19-CA-repeat homozygous men displayed a decreased risk of low-grade disease (OR = 0.50; 95% CI = 0.27-0.93), but no associations were observed with more aggressive features of disease. Additionally, there was no correlation between mean serum IGF-I protein levels and IGF-I genotype in controls. CONCLUSIONS: Further evaluation of the IGF-I CA repeat polymorphism and prostate cancer is necessary to determine if the modest risk reduction associated with the 19-CA-repeat homozygous state is observed in other study populations. PMID- 15800935 TI - Role of endothelin axis in progression to aggressive phenotype of prostate adenocarcinoma. AB - BACKGROUND: Mitogenic and anti-apoptotic actions of endothelin-1 (ET-1) are mediated through endothelin A (ET(A)) receptors. We investigated endothelin receptor expression in increasingly aggressive phenotype and in vivo effects of combination therapy using ET(A) antagonist with paclitaxel. METHODS: Dunning prostate cancer cells ranged in aggressiveness from non-tumorigenic G, to tumorigenic, non-metastatic AT-1, and to tumorigenic and metastatic MLL. Binding assays were performed alongside Q-PCR to assess receptor density. MLL xenografts were treated with vehicle, atrasentan, paclitaxel, and paclitaxel+atrasentan. RESULTS: Saturation binding assays demonstrated endothelin receptor density of MLL and AT-1 cells seven- and threefold higher than G cells, respectively. Q-PCR showed 9- and 4.5-fold greater ET(A) mRNA expression in MLL and AT-1 than G cells, respectively and no endothelin receptor B (ET(B)) expression. Combination therapy had significant effect on reduction of tumor volume than paclitaxel or atrasentan alone. CONCLUSIONS: ET(A) expression increases in aggressive prostate carcinoma. ET(A) blockade combined with paclitaxel may reduce tumor growth in advanced prostate carcinoma. PMID- 15800936 TI - BM18: A novel androgen-dependent human prostate cancer xenograft model derived from a bone metastasis. AB - BACKGROUND: Androgen-dependent prostate cancer (PrCa) xenograft models are required to study PrCa biology in the clinically relevant in vivo environment. METHODS: Human PrCa tissue from a femoral bone metastasis biopsy (BM18) was grown and passaged subcutaneously through male severe combined immune-deficient (SCID) mice. Human mitochondria (hMt), prostate specific antigen (PSA), androgen receptor (AR), cytokeratin-18 (CK-18), pan-cytokeratin, and high molecular weight cytokeratin (HMW-CK) were assessed using immunohistochemistry (IHC). Surgical castration was performed to examine androgen dependence. Serum was collected pre- and post-castration for monitoring of PSA levels. RESULTS: BM18 stained positively for hMt, PSA, AR, CK-18, pan keratin, and negatively for HMW-CK, consistent with the staining observed in the original patient material. Androgen deprivation induced tumor regression in 10/10 castrated male SCID mice. Serum PSA levels positively correlated with BM18 tumor size. CONCLUSIONS: BM18 expresses PSA and AR, and rapidly regresses in response to androgen withdrawal. This provides a new clinically significant PrCa model for the study of androgen dependent growth. PMID- 15800937 TI - Double-blind, placebo-controlled, unforced titration parallel trial of quetiapine for dopaminergic-induced hallucinations in Parkinson's disease. AB - We completed a single site, double-blind, placebo-controlled, parallel design study of quetiapine for hallucinations in PD. Thirty-one subjects with PD and prominent visual hallucinations and Mini-Mental State Examination score >21 were randomly assigned in a 2:1 drug to placebo ratio, up to 200 mg daily of quetiapine or matching placebo given in two doses. They were seen at 3 weeks (100 mg/day) and 12 weeks (200 mg/day, with optional dose reduction). Evaluation included the Unified Parkinson's Disease Rating Scale (UPDRS), the Baylor PD Hallucination Questionnaire, and a battery of neuropsychological tests. The demographics between subjects randomized to drug (n = 21) vs. placebo (n = 10) were similar. The final dose of active drug was 200 (n = 11), 150 (n = 2), 100 (n = 3), and 75 (n = 1) mg per day. All placebo subjects were on the equivalent of 200 mg per day. The UPDRS Activities of Daily Living and Motor scores did not significantly change compared to placebo. Compared to placebo, there were no significant changes in our hallucination questionnaire, the Brief Psychiatric Rating Scale (BPRS), or question 12 (hallucination item) of the BPRS. There were no significant changes on any of the neuropsychological measures. Adverse events on drug included sedation (n = 9), but no drug-related adverse events precipitated discontinuation and none were rated as serious. Quetiapine, up to 200 mg daily, was well tolerated and did not worsen UPDRS scores; however, there was no significant improvement in psychosis rating scales compared to placebo. Larger doses of drug and greater sample sizes might be considered in future studies. PMID- 15800938 TI - Interferon-gamma induces neuroendocrine-like differentiation of human prostate basal-epithelial cells. AB - BACKGROUND: Prostatic neuroendocrine (NE) cells are intraglandular hybrid epithelial-neural-endocrine cells that express and secrete numerous hormones and neuropeptides, which presumably regulate growth, differentiation, and secretory activity of the prostatic epithelium. This specialized cell type appears to differentiate from a common basal/precursor/stem cell that also gives rise to the secretory epithelium. In order to elucidate mechanisms of NE-differentiation the effects of type 1 (alpha, beta) and type 2 (gamma) interferons (IFNs) on human prostate basal cells (PrECs) were evaluated. METHODS AND RESULTS: Application of alpha/beta IFN increased the expression of the cell-cycle inhibitor p21(CIP1) and inhibited DNA synthesis, while only IFN-gamma led to increased apoptosis, cell cycle inhibitor p27(KIP1) upregulation, and differentiation of PrECs into NE-like cells. In vitro differentiated NE-like cells expressed the glycolytic enzyme neuron-specific enolase (NSE) and chromogranin A (CgA), known markers of NE-cells in vivo in the prostate. These NE-like cells also changed cytokeratin (CK) expression patterns by upregulating CK 8/18, predominantly found in terminally differentiated secretory luminal/NE epithelial cells. CONCLUSIONS: IFN-gamma produced locally in the prostate by basal cells and, under proinflammatory conditions, by infiltrating lymphocytes could support NE cell differentiation and play a role in NE differentiation processes of tumor cells in hormone-refractory prostate cancer. PMID- 15800939 TI - Connexin43 is overexpressed in Apc(Min/+)-mice adenomas and colocalises with COX 2 in myofibroblasts. AB - The expression of gap junction proteins, connexins, in the intestine and their role in tumorigenesis are poorly characterised. Truncating mutations in the tumour suppressor gene adenomatous polyposis coli (APC) are early and important events, both in inheritable (familial adenomatous polyposis, FAP) and spontaneous forms of intestinal cancer. Multiple intestinal neoplasia (Min) mice, a FAP model with inherited heterozygous mutation in Apc, spontaneously develop numerous intestinal adenomas. We recently reported reduced expression of connexin32 in Paneth cells of Min-mice. We further examine the expression of connexin43 (Cx43) and other connexins as a function of heterozygous and homozygous Apc mutation in normal intestinal tissues and adenomas of Min-mice. Qualitative analysis of connexin mRNA in intestine revealed a similar expression pattern in Min- and wild type (wt) mice. Connexin26 and connexin40 proteins were found in equal amounts in Min and wt epithelia of large and small intestine, respectively. Interestingly, the connexin43 level was increased in the stroma of Min-mice adenomas, in close proximity to epithelial cells with nuclear beta-catenin staining. Cx43 and COX-2 were located to the same areas of the adenomas, and immunostaining exhibited coexpression in the myofibroblasts. Prostaglandin E2 induces Cx43 expression and COX-2 is the rate-limiting enzyme in the prostaglandin synthesis. However, the COX-2-specific inhibitor, celecoxib, did not reduce Cx43 expression. Although both Cx43 and COX-2 are target genes for beta-catenin, they were overexpressed in stromal cells but not in epithelial tumour cells. We hypothesise that gap junctions may be of importance in the transfer of signals between epithelium and stroma. PMID- 15800940 TI - Physical activity and risk for lung cancer in a Danish cohort. AB - Physical activity might reduce the risk for lung cancer by various mechanisms, but the evidence is inconclusive. We therefore examined the relationship between physical activity and risk for lung cancer in a large population-based Danish cohort with detailed information about number of hours per week spent on specific physical activities as well as lifetime smoking patterns. Between 1993 and 1997, a total of 57,053 persons aged 50-64 years agreed to participate in the cohort. After exclusions of persons with cancer diagnosis before invitation and persons for whom data regarding study variables were missing, 26,070 men and 28,352 women remained for study. By 31 December 2002, lung cancer had been diagnosed in 194 men and 175 women. A questionnaire registered average number of hours per week spent on each of 6 types of leisure time physical activity. Level of occupational physical activity was registered in 5 categories. Cox's proportional hazard model stratified according to age at entry (1-year intervals) was adjusted for smoking, school education, possible occupational exposure to lung carcinogens and intake of fruit and vegetables. No significant association was found between number of hours per week spent on 6 types of physical activity during leisure time and the incidence rate ratio (IRR) for lung cancer. For each type of activity, the IRR of lung cancer was lower for active compared to nonactive women, whereas for men lower IRRs were only observed for sports and gardening. Higher levels of occupational physical activity had no protective effect; the lowest IRR was found for sitting work. Our study shows no convincing protective effect of physical activity on lung cancer risk. PMID- 15800941 TI - Parathyroid hormone-related protein (PTHrP) as a causative factor of cancer associated wasting: possible involvement of PTHrP in the repression of locomotor activity in rats bearing human tumor xenografts. AB - Nude rats bearing the LC-6-JCK human lung cancer xenograft displayed cancer associated wasting syndrome in addition to humoral hypercalcemia of malignancy. In these rats, not only PTHrP but also several other human proinflammatory cytokines, such as IL-6, leukemia-inducing factor, IL-8, IL-5 and IL-11, were secreted to the bloodstream. Proinflammatory cytokines induce acute-phase reactions, as evidenced by a decrease of serum albumin and an increase in alpha1 acid glycoprotein. Tumor resection abolished the production of proinflammatory cytokines and improved acute-phase reactions, whereas anti-PTHrP antibody affected neither proinflammatory cytokine production nor acute-phase reactions. Nevertheless, tumor resection and administration of anti-PTHrP antibody similarly and markedly attenuated not only hypercalcemia but also loss of fat, muscle and body weight. Body weight gain by anti-PTHrP antibody was associated with increased food consumption; increased body weight from anti-PTHrP antibody was observed when animals were freely fed but not when they were given the same feeding as those that received only vehicle. Furthermore, nude rats bearing LC-6 JCK showed reduced locomotor activity, less eating and drinking and low blood phosphorus; and anti-PTHrP antibody restored them. Although alendronate, a bisphosphonate drug, decreased blood calcium, it affected neither locomotor activity nor serum phosphorus level. These results indicate that PTHrP represses physical activity and energy metabolism independently of hypercalcemia and proinflammatory cytokine actions and that deregulation of such physiologic activities and functions by PTHrP is at least in part involved in PTHrP-induced wasting syndrome. PMID- 15800942 TI - Migration-stimulating factor displays HEXXH-dependent catalytic activity important for promoting tumor cell migration. AB - Like most extracellular matrix (ECM) components, fibronectin (Fn) is proteolyzed generating specific activities. Fibronectin proteinase (Fn-proteinase) represents such a cryptic activity located in the gelatin-binding domain (GBD) of Fn and displays a zinc metalloproteinase activity. The migration-stimulating factor (MSF) is a truncated Fn isoform generated by alternative mRNA splicing and corresponds to the N-terminal part of Fn that comprises the GBD. We show that several human mammary epithelial cells express MSF and constitutively produce Fn proteinase activity. Furthermore, recombinant MSF produced by HEK-293 and MCF-7 cells possesses a constitutive Fn-proteinase activity. Mutating the putative zinc binding motif, HEXXH, of the protein abolishes its activity thereby demonstrating its specificity. Using PCR, we showed that MSF is barely expressed in normal breast tissues, whereas its expression is significantly increased in tumors. Furthermore, an association between MSF expression and invasive capacity is observed in various breast adenocarcinoma cell lines. Indeed, when stably transfected in non-invasive MCF-7 cells, MSF promotes cell migration in a mechanism mostly dependent on its Fn-proteinase activity. In summary, our study shows that: (i) MSF displays constitutive Fn-proteinase activity; (ii) MSF expression is induced in human breast cancer; and (iii) MSF confers pro-migratory activity that depends mostly on its Fn-proteinase activity. These results suggest that MSF may be involved in tumor progression. PMID- 15800943 TI - Case-control study of familial lung cancer risks in UK women. AB - Family history data from a case-control study of lung cancer conducted in the United Kingdom between 1999 and 2004 were analysed to estimate familial risks of the disease. Comparison of lung cancer prevalence in first-degree relatives of 1,482 female lung cancer cases and 1,079 female controls was undertaken using logistic regression adjusting for age and tobacco exposure. Overall, lung cancer in a first-degree relative was associated with a significant increase in the risk of lung cancer [odds ratio (OR) 1.49; 95% confidence interval (CI), 1.13-1.96]. For cases with early onset of the disease (< 60 years), the OR of lung cancer was 2.02 (95% CI, 1.22-3.34). Having 2 or more affected relatives was associated with an OR of 2.68 (95% CI, 1.29-5.55), with a significant trend in risk according to the number of relatives affected (p = 0.001). An increased risk of lung cancer associated with family history of the disease was observed when analysis was restricted to lifetime nonsmokers, although this did not reach significance (OR 1.23; 95% CI, 0.65-2.31). Results confirm previous findings and support the role of a familial predisposition to lung cancer. PMID- 15800944 TI - Herceptin-induced inhibition of ErbB2 signaling involves reduced phosphorylation of Akt but not endocytic down-regulation of ErbB2. AB - The anti-proliferative effect of the ErbB2 specific antibody Herceptin in cells overexpressing ErbB2 has previously been explained by endocytic downregulation of ErbB2. However, in the following, we demonstrate that Herceptin inhibited proliferation of ErbB2 overexpressing cells without downregulating ErbB2. Herceptin did also not induce endocytosis of ErbB2. Herceptin was found to blunt proliferation of SKBr3 cells overexpressing EGFR, ErbB2, and ErbB3 and expressing functional PTEN, probably by recruiting PTEN to the plasma membrane. Akt was found to be constitutively phosphorylated both in SKBr3 cells overexpressing EGFR, ErbB2 and ErbB3, and in SKOv3 cells, overexpressing EGFR and ErbB2. However, phosphorylation of Akt was inhibited by Herceptin only in SKBr3 cells. SKOv3 cells, which lack the tumour suppressor protein Ras homolog member I, was found to have constitutively phosphorylated mitogen activated protein kinase and functionally increased Ras activity. SKOv3 cells further had low expression levels of PTEN. We thus confirm that the anti-proliferative effect of Herceptin in SKBr3 cells is due to recruitment of PTEN to the plasma membrane and conclude that Herceptin does not blunt phosphatidyl inositol 3 kinase-induced growth in cells with constitutive Ras activity. We further conclude that endocytic downregulation of ErbB2 does not contribute to Herceptin's antiproliferative effect. PMID- 15800945 TI - Palifermin reduces diarrhea and increases survival following irinotecan treatment in tumor-bearing DA rats. AB - Mucositis is a common side effect of cancer chemotherapy for which there is currently no treatment. Irinotecan is a commonly used effective chemotherapeutic agent, causing severe gastrointestinal mucositis and diarrhea. Previous research suggests that palifermin is potentially antimucotoxic. The primary aim of this study was to determine whether palifermin was effective in ameliorating irinotecan-induced gastrointestinal mucositis. We also determined the protective effects of single large and multiple small doses of palifermin. Tumor-bearing DA rats were treated with a single large (10 mg/kg) dose of palifermin 3 days prior to, or multiple small (3 mg/kg day for 3 days) doses of palifermin or vehicle control prior to, receiving 2 doses of 150 mg/kg irinotecan. Animals were killed at 6, 24, 48, 72, 96, 120, or 144 hr after treatment. The primary endpoints were diarrhea and mortality. Gastrointestinal morphometry, histopathology and apoptosis were assessed. Tumor weights and mitoses were measured to ensure palifermin did not promote tumor growth. Data were analyzed using Peritz' F-test, Student's t-test and Tukey-Kramer multiple comparison test. Animals pretreated with palifermin tolerated irinotecan treatment better than control animals with less severe diarrhea (5% in animals receiving 10 mg/kg palifermin, 11% in animals receiving 3 x 3 mg/kg palifermin and 28% in animals receiving irinotecan only) and reduced mortality (2% in animals receiving 10 mg/kg palifermin, 11% in animals receiving 3 x 3 mg/kg palifermin and 28% in animals receiving irinotecan only). Small and large intestinal weights were maintained. Intestinal morphometry was not maintained in palifermin-pretreated rats despite being increased prior to irinotecan treatment. Palifermin pretreatment did not prevent apoptosis that peaked at 6 hr in the jejunum or colon, but prevented apoptosis at 96 hr in the small intestine. Palifermin pretreatment in both treatment regimens significantly reduces diarrhea and mortality following irinotecan administration without adversely affecting tumor growth. This positive response warrants further investigation, particularly in humans. PMID- 15800946 TI - Polymorphisms of XRCC1 gene, alcohol consumption and colorectal cancer. AB - To evaluate contribution of polymorphisms of the XRCC1 gene to the risk of colorectal cancer, we conducted a case-control study of 209 colorectal cancer cases and 209 age- and gender-matched controls in the Korean population. We tested the hypothesis by constructing allele combinations with known SNP. Allelic variants of the XRCC1 gene at codons 194, 280 and 399 were analyzed in lymphocyte DNA by PCR-RFLP. We observed an increased risk of colorectal cancer associated with the 399Gln allele. The odds ratio (OR) was 1.61 (95% confidence interval [CI] 1.09-2.39) for the 399Gln allele. When combined allele-specific OR were calculated after estimating frequencies, 3 common allele combinations were found to be associated with an increased risk of colorectal cancer. The OR for the 194Trp-280Arg-399Arg was 1.48 (95% CI = 1.06-2.07) using 194Arg-280Arg-399Arg as the reference. The OR for the 194Arg-280His-399Arg and the 194Arg-280Arg-399Gln were 1.78 (95% CI = 1.09-2.89) and 1.78 (95% CI = 1.23-2.59), respectively. Analysis after controlling for smoking, exercise and dietary habits indicated that alcohol consumption (> or =80 g/week) is a significant risk factor of colorectal cancer (OR = 2.60, 95% CI = 1.46-4.62). An increased risk for colorectal cancer was identified in alcohol drinkers with the risky allele combinations. Our results suggest that polymorphisms in the XRCC1 genes may contribute to colorectal cancer susceptibility, and some evidence was obtained of a genetic modification for the relationship between alcohol intake and colorectal cancer. PMID- 15800947 TI - Induction of autoantibodies to syngeneic prostate-specific membrane antigen by xenogeneic vaccination. AB - Prostate-specific membrane antigen (PSMA) is a prototypical differentiation antigen expressed on normal and neoplastic prostate epithelial cells, and on the neovasculature of many solid tumors. Monoclonal antibodies specific for PSMA are in development as therapeutic agents. Methodologies to actively immunize against PSMA may be limited by immunologic ignorance and/or tolerance that restrict the response to self-antigens. Our studies have previously shown that xenogeneic immunization with DNA vaccines encoding melanosomal differentiation antigens induces immunity in a mouse melanoma model. Here we apply this approach to PSMA to establish proof of principle in a mouse model. Immunization with xenogeneic human PSMA protein or DNA induced antibodies to both human and mouse PSMA in mice. Monoclonal antibodies specific for mouse PSMA were generated to analyze antibody isotypes and specificity for native and denatured PSMA at the clonal level. Most antibodies recognized denatured PSMA, but C57BL/6 mice immunized with xenogeneic PSMA DNA followed by a final boost with xenogeneic PSMA protein yielded autoantibodies that reacted with native folded mouse PSMA. Monoclonal antibodies were used to confirm the expression of PSMA protein in normal mouse kidney. These results establish the basis for clinical trials to test PSMA DNA vaccines in patients with solid tumors that either express PSMA directly or that depend on normal endothelial cells expressing PSMA for their continued growth. PMID- 15800948 TI - Ewing tumours and synovial sarcomas have critical features of karyotype evolution in common with epithelial tumours. AB - We have analysed the accumulated cytogenetic data on karyotypic evolution in Ewing tumours (ET) and synovial sarcomas (SS). Both tumour types frequently show balanced translocations, t(11;22) and t(X;18), respectively, that result in specific fusion genes. The analyses revealed +8, +12, +1q, and 16q- as important secondary changes to t(11;22) in ET and the imbalances showed a distinct temporal order. By principal component analysis, one major karyotypic pathway dominated by gains and one minor dominated by losses were identified. The kartyotypic evolution pattern in SS was less distinct. Both ET and SS showed a power law distribution of the number of acquired aberrations, which in both tumour types conformed to a distribution with an exponent equal to 1. Similar distributions are frequently found in epithelial tumours. ET and SS differ in this respect from other malignancies with balanced translocations resulting in fusion genes, which typically show a power law distribution of the number of acquired aberrations with exponents close to 2. This suggests that chromosome changes in ET and SS may develop through mechanisms more similar to those in epithelial tumours lacking recurrent balanced rearrangements than in haematological malignancies characterised by balanced translocations leading to fusion genes. PMID- 15800949 TI - Incidence differences for gallbladder cancer between occupational groups suggest an etiological role for alcohol. PMID- 15800950 TI - Aerosol gemcitabine inhibits the growth of primary osteosarcoma and osteosarcoma lung metastases. AB - Osteosacarcoma (OS) lung metastases are often resistant to chemotherapy. Most anticancer drugs are administered systemically. In many cases this is followed by dose-dependent toxicity, which may not allow the achievement of therapeutic levels in lungs to eradicate metastases. We determined the efficacy of gemcitabine (GCB) by administering it directly to the lungs via aerosol and studied the role of the Fas pathway in response to the therapy. We used 2 osteosarcoma lung metastases animal models: human LM7 cells that form lung metastases in mice following intravenous injection and murine LM8 cells, which grows subcutaneously in mice and spontaneously metastasize to the lung. Treatment was initiated when the presence of lung metastases had been established. Aerosol GCB inhibited the growth of lung metastases in mice. Intraperitoneal GCB administration at similar dosage had no effect on lung metastases. Besides its direct effect on lung metastases, aerosol GCB suppressed the growth of subcutaneous LM8 tumor. Histopathological examination of mice receiving aerosol GCB showed no evidence of toxicity. Lungs are distinguished from other tissues by the constitutive expression of FasL. Since exposure of tumor cells to GCB upregulated Fas expression, we hypothesized that the susceptibility of the tumor cells to ligand-induced cell death by resident lung cells may be increased. Therefore, the Fas pathway may contribute to the therapeutic effect of aerosol GCB. PMID- 15800951 TI - Serum cytochrome c indicates in vivo apoptosis and can serve as a prognostic marker during cancer therapy. AB - Despite significant progress in cancer therapy, the outcome of the treatment is often unfavorable. Better treatment monitoring would not only allow an individual more effective, patient-adjusted therapy, but also it would eliminate some of the side effects. Using a cytochrome c ELISA that was modified to increase sensitivity, we demonstrate that serum cytochrome c is a sensitive apoptotic marker in vivo reflecting therapy-induced cell death burden. Furthermore, increased serum cytochrome c level is a negative prognostic marker. Cancer patients whose serum cytochrome c level was normal 3 years ago have a twice as high probability to be still alive, as judged from sera samples collected for 3 years, analyzed recently and matched with survival data. Moreover, we show that serum cytochrome c and serum LDH-activity reflect different stages and different forms of cell death. Cellular cytochrome c release is specific for apoptosis, whereas increased LDH activity is an indicator of (secondary) necrosis. Whereas serum LDH activity reflects the "global" degree of cell death over a period of time, the sensitive cytochrome c-based method allows confirmation of the individual cancer therapy-induced and spontaneous cell death events. The combination of cytochrome c with tissue-specific markers may provide the foundation for precise monitoring of apoptosis in vivo, by "lab-on-the-chip" technology. PMID- 15800952 TI - Phospholipid polymer surfaces reduce bacteria and leukocyte adhesion under dynamic flow conditions. AB - Persistence of infection can occur when the host immune response is compromised because of the presence of a foreign implant. Surface modification of biomaterials with phospholipid polymers may enhance biocompatibility and reduce incidence of infection by impeding bacterial and leukocyte adhesion. A rotating disk model, which generates shear stress from 0 to 18 dynes/cm(2), was used to characterize adhesion of neutrophils, monocytes, and bacteria in phosphate buffered saline (PBS) or 25% human serum on polyethylene terephthalate surfaces coated with a phospholipid polymer, poly[omega-methacryloyloxyalkyl phosphorylcholine (MAPC)-co-n-butyl methacrylate (BMA)]. The material designated PMB30 contains a methylene chain length, (CH(2))(n), of n = 2, whereas PMHB30 contains a chain length of n = 6. In PBS, bacterial adhesion was shear stress dependent with the lowest bacterial density observed on PMB30. However, the presence of serum proteins eliminated shear stress and surface chemistry effects in addition to bacterial adhesion reduced to <10% of adhesion in PBS. Trends for leukocyte adhesion in serum demonstrated shear dependence with PMB30 exhibiting the lowest cell density throughout the range of shear stresses. In conclusion, modification of the polyethylene terephthalate surfaces with phospholipid polymers resulted in reduced bacterial and leukocyte adhesion. Furthermore, shortening the methylene chain length of the MAPC copolymer most effectively reduced adhesion. PMID- 15800953 TI - The role of primary repair for colonic injuries in wartime. AB - BACKGROUND: The study reviewed an experience of selective primary repair for penetrating colonic injuries incorporating a number of procedures during the 1992 1995 Bosnia-Herzegovina conflict. METHODS: Of 5370 casualties, 259 (4.8 per cent) had injuries to the colon. The patients were divided into two groups: those who had primary repair and those who needed a colostomy. The patients' records were reviewed to determine the cause of injury (explosive weapons or bullets), the position and type of colon injury, associated injuries, the surgical procedure(s) done, complications related to the colonic wound or its management, and mortality. RESULTS: Some 122 (47.1 per cent) patients had primary colonic repair and 137 (52.9 per cent) had a colostomy. One hundred and fifty (57.9 per cent) were injured by explosive weapons, 108 (41.7 per cent) had bullet wounds and one (0.4 per cent) a stab injury. Associated injuries were seen in 249 (96.1 per cent) patients. Complications related to the colonic wound or its management developed in 27 per cent of patients after primary repair and 30 per cent after colostomy. Mortality rates were 8.2 per cent and 7.2 per cent, respectively. CONCLUSION: Primary repair was a safe and effective treatment for penetrating colonic injuries during war. PMID- 15800954 TI - Laparoscopic repair of large hiatal hernias. AB - BACKGROUND: The repair of large hiatal hernias can be technically challenging. Most series describing laparoscopic repair report only symptomatic outcomes and the true recurrence rate, including asymptomatic recurrence, is not well documented. This study evaluated the long-term outcome of laparoscopic repair of large hiatal hernias. METHODS: All patients who had undergone laparoscopic repair of a large hiatus hernia (more than 50 per cent of the stomach in the hernia) with a minimum 2-year clinical follow-up were identified from a prospectively maintained database. A standardized questionnaire was used to assess symptoms and a barium swallow radiograph was performed to determine anatomy. Multivariate analysis was used to identify factors associated with recurrence. RESULTS: Of 100 eligible patients, clinical follow-up was available in 96. Follow-up ranged from 2 to 8 (median 4) years. In patients with preoperative reflux symptoms, there were significant improvements in heartburn and dysphagia scores after surgery. Overall, 80 per cent of patients rated their outcome as good or excellent. Sixty patients underwent a postoperative barium meal examination that identified 14 radiological hernia recurrences (eight small, three medium and three large). Four other patients in this group of 60 had previously undergone reoperation for early and late recurrence (two of each), giving an overall recurrence rate of 18 of 60 (30 per cent). One third of patients with recurrence were totally asymptomatic and the presence of postoperative symptoms did not reliably predict the presence of anatomical recurrence. Younger age and increased weight at operation were independent risk factors contributing to recurrence. CONCLUSIONS: Laparoscopic repair of large hiatal hernias yields good clinical outcome. Recurrence after laparoscopic repair seems to be more common than previously thought. Objective anatomical studies are required to determine the true recurrence rate. The majority of recurrences are not large and do not cause significant symptoms. PMID- 15800955 TI - Validity of the Glasgow Aneurysm Score and the Hardman Index in predicting outcome after ruptured abdominal aortic aneurysm repair. AB - BACKGROUND: The Glasgow Aneurysm Score and the Hardman Index have been recommended as predictors of outcome after repair of ruptured abdominal aortic aneurysm (AAA). This study aimed to assess their validities. METHODS: Patients admitted to a single unit with a ruptured AAA over a 2-year interval (2000-2001) were identified from a prospectively compiled database. Hospital records of all patients undergoing attempted operative repair were reviewed. The Glasgow Aneurysm Score and the Hardman Index were calculated retrospectively and related to clinical outcome. RESULTS: One hundred patients were admitted with a ruptured AAA. Of these, 82 underwent attempted operative repair and were included in the study: 68 men and 14 women, of median age 73 (range 54-87) years. Thirty (37 per cent) patients died after the operation. The Glasgow Aneurysm Score was a poor predictor of postoperative mortality. The area under the Receiver-Operator Characteristic curve was 0.606 (P = 0.112, 95 per cent c.i. 0.483-0.729). Similarly, the Hardman Index failed to predict postoperative mortality accurately (P = 0.211, chi(2) for trend). Of nine patients in this series with three or more Hardman criteria, generally held to be fatal, six survived. CONCLUSION: Contrary to previous reports, The Glasgow Aneurysm Score and the Hardman Index were poor predictors of postoperative mortality after repair of a ruptured AAA in this study. PMID- 15800956 TI - HIV coinfection shortens the survival of patients with hepatitis C virus-related decompensated cirrhosis. AB - The impact of human immunodeficiency virus (HIV) coinfection on the survival of patients with hepatitis C virus (HCV)-related end-stage liver disease (ESLD) is unknown. Because HIV infection is no longer considered an absolute contraindication for liver transplantation in some countries, it has become a priority to address this topic. The objective of this study was to compare the survival of HIV-infected and HIV-uninfected patients with decompensated cirrhosis due to HCV. In a retrospective cohort study, the survival of 1,037 HCV monoinfected and 180 HCV/HIV-coinfected patients with cirrhosis after the first hepatic decompensation was analyzed. Of the group, 386 (37%) HCV-monoinfected and 100 (56%) HCV/HIV-coinfected subjects died during the follow-up. The median survival time of HIV-infected and HIV-uninfected patients was 16 and 48 months, respectively (P < .001). The relative risk (95% CI) of death for HIV-infected patients was 2.26 (1.51-3.38). Other independent predictors of survival were age older than 63 years (2.25 [1.53-3.31]); Child-Turcotte-Pugh class B versus class A (1.95 [1.41-2.68]) and class C versus class A (2.78 [1.66-4.70]); hepatitis D virus infection (1.56 [1.12-4.77]); model for end-stage liver disease score, (1.05 [1.01-1-11]); more than one simultaneous decompensation (1.23 [1.12-3.33]); and the type of the first hepatic decompensation, with a poorer prognosis associated with encephalopathy compared with portal hypertensive gastrointestinal bleeding (2.03 [1.26-3.10]). In conclusion, HIV coinfection reduces considerably the survival of patients with HCV-related ESLD independently of other markers of poor prognosis. This fact must be taken into account to establish the adequate timing of liver transplantation in HIV-coinfected subjects. PMID- 15800957 TI - Evaluation of gestational age and admission date assumptions used to determine prenatal drug exposure from administrative data. AB - OBJECTIVE: Our aim was to evaluate the 270-day gestational age and delivery date assumptions used in an administrative dataset study assessing prenatal drug exposure compared to information contained in a birth registry. STUDY DESIGN AND SETTING: Kaiser Permanente Colorado (KPCO), a member of the Health Maintenance Organization (HMO) Research Network Center for Education and Research in Therapeutics (CERTs), previously participated in a CERTs study that used claims data to assess prenatal drug exposure. In the current study, gestational age and deliveries information from the CERTs study dataset, the Prescribing Safely during Pregnancy Dataset (PSDPD), was compared to information in the KPCO Birth Registry. Sensitivity and positive predictive value (PPV) of the claims data for deliveries were assessed. The effect of gestational age and delivery date assumptions on classification of prenatal drug exposure was evaluated. RESULTS: The mean gestational age in the Birth Registry was 273 (median = 275) days. Sensitivity of claims data at identifying deliveries was 97.6%, PPV was 98.2%. Of deliveries identified in only one dataset, 45% were related to the gestational age assumption and 36% were due to claims data issues. The effect on estimates of prevalence of prescribing during pregnancy was an absolute change of 1% or less for all drug exposure categories. For Category X, drug exposures during the first trimester, the relative change in prescribing prevalence was 13.7% (p = 0.014). CONCLUSION: Administrative databases can be useful for assessing prenatal drug exposure, but gestational age assumptions can result in a small proportion of misclassification. PMID- 15800958 TI - Randomized clinical trial of pylorus-preserving duodenopancreatectomy versus classical Whipple resection-long term results. AB - BACKGROUND: It is not known whether pylorus-preserving duodenopancreatectomy is as effective as the classical Whipple procedure in the resection of pancreatic and periampullary tumours. A prospective randomized trial was undertaken to compare the results of the two procedures. METHODS: Clinical data, histological findings, short-term results, survival and quality of life of all patients having surgery for suspected pancreatic or periampullary cancer between June 1996 and September 2001 were analysed. RESULTS: Two hundred and fourteen patients were randomized to undergo either a standard or a pylorus-preserving Whipple resection. After exclusion of 84 patients on the basis of intraoperative findings, 130 patients (66 standard Whipple operation and 64 pylorus-preserving resection) were entered into the trial. Of these, 110 patients with proven adenocarcinoma (57 standard Whipple and 53 pylorus-preserving resection) were analysed for long-term survival and quality of life. There was no difference in perioperative morbidity. Long-term survival, quality of life and weight gain were identical after a median follow-up of 63.1 (range 4-93) months. At 6 months, capacity to work was better after the pylorus-preserving procedure (77 versus 56 per cent; P = 0.019). CONCLUSION: Both procedures were equally effective for the treatment of pancreatic and periampullary cancer. Pylorus-preserving Whipple resection offers some minor advantages in the early postoperative period, but not in the long term. PMID- 15800959 TI - Vibrational infrared conformational studies of model peptides representing the semicrystalline domains of Bombyx mori silk fibroin. AB - The structural organization of Bombyx mori silk fibroin was investigated by infrared (IR) spectroscopy. To this aim, (AG)15 and other model peptides of varying chain length, containing tyrosine (Y), valine (V), and serine (S) in the basic (AG)n sequence were synthesized by the solid phase method and their spectroscopic properties were determined. Both the position and the relative content of Y, V, and S residues in the (AG)n model system appeared critical in determining the preferred conformation, i.e., silk I, silk II, and unordered structures. Curve fitting analysis in the amide I range showed that the model peptides with prevailing silk II structure displayed different beta-sheet content, which was dependent on the degree of interruption of the (AG)n sequence. In this regard, the bands at about 1000 and 980 cm(-1), specifically assigned to the AG sequence of the B. mori silk fibroin chain, were identified as marker of the degree of interruption of the (AG)n sequence.A stable silk I structure was observed only when the Y residue was located near the chain terminus, while a silk I --> silk II conformational transition occurred when it was positioned in the central region of the peptide. Analysis of the second-derivative spectra in the amide I range allowed us to identify a band at 1639 cm(-1) (4 --> 1 hydrogen bonded type II beta-turns), which is characteristic of the silk I conformation. PMID- 15800960 TI - Chip-based microfluidic devices coupled with electrospray ionization-mass spectrometry. AB - We present the current status of the development of microfluidic devices fabricated on different substrates for coupling with electrospray ionization-mass spectrometry (ESI-MS). Until now, much success has been gained in fabricating the ESI chips, which show better performances due to miniaturization when compared with traditional methods. Integration of multiple steps for sample preparation and ESI sample introduction, however, remains a great challenge. This review covers the main technical development of electrospray device that were published from 1997 to 2004. This article does not attempt to be exclusive. Instead, it focuses on the publications that illustrated the breath of the development and applications of microchip devices for MS-based analysis. PMID- 15800961 TI - Detection of a novel familial catalase mutation (Hungarian type D) and the possible risk of inherited catalase deficiency for diabetes mellitus. AB - The enzyme catalase is the main regulator of hydrogen peroxide metabolism. Recent findings suggest that a low concentration of hydrogen peroxide may act as a messenger in some signalling pathways whereas high concentrations are toxic for many cells and cell components. Acatalasemia is a genetically heterogeneous condition with a worldwide distribution. Yet only two Japanese and three Hungarian syndrome-causing mutations have been reported. A large-scale (23 130 subjects) catalase screening program in Hungary yielded 12 hypocatalasemic families. The V family with four hypocatalasemics (60.6 +/- 7.6 MU/L) and six normocatalasemic (103.6 +/- 23.5 MU/L) members was examined to define the mutation causing the syndrome. Mutation screening yielded four novel polymorphisms. Of these, three intron sequence variations, namely G-->A at the nucleotide 60 position in intron 1, T-->A at position 11 in intron 2, and G-->T at position 31 in intron 12, are unlikely to be responsible for the decreased blood catalase activity. However, the novel G-->A mutation in exon 9 changes the essential amino acid Arg 354 to Cys 354 and may indeed be responsible for the decreased catalase activity. This inherited catalase deficiency, by inducing an increased hydrogen peroxide steady-state concentration in vivo, may be involved in the early manifestation of type 2 diabetes mellitus for the 35-year old proband. PMID- 15800962 TI - Isoelectric focusing in cyclic olefin copolymer microfluidic channels coated by polyacrylamide using a UV photografting method. AB - As an alternative material to glass or silicon, microfluidic devices made from a cyclic olefin copolymer (COC) were fabricated. This material is of interest because of the relative ease of fabrication, low costs, and solvent resistance. However, as a result of the strong hydrophobic interactions normally present, COC surfaces are not suitable for protein separations. To reduce the protein adsorption and make COC suitable for protein separations, UV-initiated grafting of polyacrylamide was used to coat the surface of COC devices. The change in surface properties caused by different graft times was studied. The surface hydrophilicity and electroosmotic mobility were characterized by contact angle and electroosmosis measurements. Isoelectric focusing was performed to test protein separations in polyacrylamide-coated COC microchannels. A single protein, carbonic anhydrase, was used to analyze the focusing effects and peak capacities in uncoated and polyacrylamide-coated COC devices. Peak capacities ranging from 75 to 190 were achieved with a polyacrylamide-coated surface. A mixture of two proteins, conalbumin labeled with Alexa Fluor 488 and beta-lactoglobulin A labeled with Alexa Fluor 546, was used to test protein separations. Linear and rapid separation of proteins was achieved in the polyacrylamide-coated COC microfluidic device. PMID- 15800963 TI - Tris(2,2'-bipyridyl)ruthenium(II)-zirconia-Nafion composite films applied as solid-state electrochemiluminescence detector for capillary electrophoresis. AB - The major goal of this work was to develop a new solid-state electrochemiluminescence (ECL) detector suitable for capillary electrophoresis (CE). The detector was fabricated by coating a sol-gel derived zirconia (ZrO(2)) Nafion composite film on a graphite electrode, then the zirconia-Nafion modified electrode was immersed in tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)(3) (2+)) solution to immobilize this active chemiluminescence reagent. The voltammetric and ECL behaviors of the detector were investigated and optimized in tripropylamine solution. The ratio of 53% for zirconia in the zirconia-Nafion composite provided the highest luminescence intensity of immobilized Ru(bpy)(3) (2+). The ECL can maintain its stability very well in the phosphate solution in the period of 5-90 h when the solid-state ECL detector was immersed in the solution all the time. The optimum distance of capillary outlet to the solid state ECL detector has been found to be ca. 50-80 microm for a 75 microm capillary. The effects of ionic strength and pH of ECL solution on peak height were investigated. The CE with solid-state ECL detector system was successfully used to detect tripropylamine, lidocaine, and proline. The detection limits (S/N = 3) were 5 x 10(-9) mol.L(-1) for tripropylamine, 1 x 10(-8) mol.L(-1) for lidocaine and 5 x 10(-6) mol.L(-1) for proline, and the linear ranges were from 1.0 x 10(-8) to 1.0 x 10(-5) mol.L(-1) for tripropylamine, 5.0 x 10(-7) mol.L(-1) to 1.0 x 10(-5) mol.L(-1) for lidocaine and 1.0 x 10(-5) to 1.0 x 10(-3) mol.L( 1) for proline, respectively. PMID- 15800964 TI - A generic approach to the impurity profiling of drugs using standardised and independent capillary zone electrophoresis methods coupled to electrospray ionisation mass spectrometry. AB - Three standardised, capillary zone electrophoresis-electrospray ionisation mass spectrometry (CZE-ESI-MS) methods were developed for the analysis of six drug candidates and their respective process-related impurities comprising a total of 22 analytes with a range of functional groups and lipophilicities. The selected background electrolyte conditions were found to be: 60/40 v/v 10 mM ammonium formate pH 3.5/organic, 60/40 v/v 10 mM ammonium acetate pH 7.0/organic and 10 mM piperidine, pH 10.5, where the organic solvent is 50/50 v/v methanol/acetonitrile. The coaxial sheath flow consisted of either 0.1% v/v formic acid in 50/50 v/v methanol/water, or 10 mM ammonium acetate in 50/50 v/v methanol/water, depending on the mixture being analysed. Factor analysis and informational theory were used to quantify the orthogonality of the methods and predict their complementarities. The three selected CZE-ESI-MS methods allowed the identification of 21 out of 22 of all the drug candidates and their process related impurities and provided orthogonality with four established high performance liquid chromatography-mass spectrometry (HPLC-MS) methods. These methodologies therefore form the basis of a generic approach to impurity profiling of pharmaceutical drug candidates and can be applied with little or no analytical method development, thereby offering significant resource and time savings. PMID- 15800965 TI - Cyclical electrical field flow fractionation. AB - Cyclical electrical field flow fractionation (Cy/ElFFF) is demonstrated in a standard electrical field flow fractionation (ElFFF) channel for the first time. Motivation for the use of alternating current (AC) fields in a traditionally direct current (DC) technique are discussed. The function of the system over a wide range of operating conditions is explored and challenges associated with various operating conditions reported. Retention of polystyrene nanoparticle standards is accomplished and the effect of varying parameters of the applied field, such as voltage and frequency, are explored. The first separations using this technique are demonstrated. The experimental results are compared to analytical models previously reported in the literature. The general trend of the experimental results is similar to those predicted in theoretical models and possible reasons for discrepancies are elucidated. Suggestions are made for improving the separation and analysis method, and possible applications explored. PMID- 15800966 TI - Capillary electrophoresis hyphenated to inductively coupled plasma-sector field mass spectrometry for the detection of chromium species after incubation of chromium in simulated sweat. AB - The presence of chromium in chromium-tanned leather represents a considerable health problem since it can lead to chronic allergic contact dermatitis. Apart from trivalent chromium (Cr(III)), which is used for tanning, leather often contains hexavalent chromium (Cr(VI)), resulting from the oxidation of Cr(III) during the tanning process. This study deals with the chromium compounds in simulated sweat when brought into contact with Cr(III) or Cr(VI) and with chromium-tanned leathers. A capillary electrophoresis (CE) method was developed, with inductively coupled plasma-sector field-mass spectrometry (ICP-SF-MS) for element-specific detection. Two different electrophoretic runs, applying once the positive and once the negative polarity mode, were necessary for the detection of positively and negatively charged chromium species. Although sometimes described in the literature, a pre-run derivatization of the chromium-species was not performed here to prevent species transformation. 50 mmol.L(-1) sodium phosphate at a pH of 2.5 was used as CE separation buffer and as make-up liquid for the CE ICP-SF-MS interface. When applied to simulated sweat samples incubated with Cr(VI), this method showed that methionine is responsible for the reduction of Cr(VI) into Cr(III), which, at its turn, forms a complex with lactic acid. In the case of sweat plus Cr(III), the latter step was also seen. Applied to simulated sweat in contact with leather samples, the method developed showed the presence of the former species among a much more complex pattern. PMID- 15800967 TI - Analysis of positional isomers of hydroxylated aromatic cytokinins by micellar electrokinetic chromatography. AB - A micellar electrokinetic chromatography (MEKC) method was developed for the separation of six positional isomers of hydroxylated aromatic cytokinins (topolin and topolin riboside), including ortho-topolin, meta-topolin, para-topolin, ortho topolin riboside, meta-topolin riboside, and para-topolin riboside. Optimum resolution and analysis time (ca. 20 min) for the six aromatic cytokinin standards were achieved with a running buffer at pH 8.0 consisting of 20 mM boric acid, 50 mM sodium dodecyl sulfate (SDS), and 20% v/v methanol. The method has good reproducibility and has been successfully applied to detect the presence of a putative ortho-topolin in coconut water extract sample purified using C(18) and mixed-mode solid-phase extraction (SPE) columns. Other advantages of this MEKC method are short analysis time, low solvent consumption, and separation of positional isomers which could be achieved by a simple aqueous buffer system without the use of expensive chromatographic columns. In addition, a high performance liquid chromatography (HPLC) method with baseline separation of the six topolin and topolin riboside standards was developed for the confirmation of the endogenous ortho-topolin in coconut water sample. Finally, the presence of ortho-topolin was further confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) based on its characteristic fragmentation pattern. PMID- 15800968 TI - The first two-dimensional reference map of the fission yeast, Schizosaccharomyces pombe proteins. AB - Cytosolic proteins of Schizosaccharomyces pombe were separated by two-dimensional (2-D) gel electrophoresis, to construct the first 2-D reference map. In the pI range 4-7, more than 500 spots were detected by silver staining, and 70 different proteins corresponding to 111 spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and tandem mass spectrometry, where necessary. In the pI range 6-9, approximately 330 spots were detected, and 31 proteins corresponding to 38 spots were identified by mass spectrometry. More than 50% of the identified proteins were involved in amino acid, carbohydrate or nucleotide metabolism, and energy production. A second large group of identified proteins comprises heat shock and other stress related proteins and chaperones. PMID- 15800969 TI - Simultaneous determination of anthraquinones in Rhubarb by pressurized liquid extraction and capillary zone electrophoresis. AB - Rhubarb, a well-known Chinese herbal medicine, is also used in Europe and other places of the world. Anthraquinones derivatives are thought to be the major active components. A pressurized liquid extraction (PLE) and capillary zone electrophoresis (CZE) separation were developed for simultaneous determination of five anthraquinones including aloe-emodin, emodin, chrysophanol, physcion, and rhein in Rhubarb. The effects of the experimental variables on PLE and CZE have been optimized. The optimum conditions of PLE were: solvent, methanol; temperature, 140 degrees C; particle size, 0.13-0.2 mm; static extraction time, 5 min; pressure, 1500 psi; and one extraction. The best separation of the five anthraquinones could be obtained using 50 mM borate buffer (pH 8.2) containing 25% isopropyl alcohol and 25% acetontrile as modifier, while the separation voltage was 25 kV and the temperature was at 20 degrees C. The method developed is accurate, simple, and reproducible, and could be used for quality control of Rhubarb and its medical preparations. PMID- 15800970 TI - Large-scale identification of proteins in human salivary proteome by liquid chromatography/mass spectrometry and two-dimensional gel electrophoresis-mass spectrometry. AB - Human saliva contains a large number of proteins and peptides (salivary proteome) that help maintain homeostasis in the oral cavity. Global analysis of human salivary proteome is important for understanding oral health and disease pathogenesis. In this study, large-scale identification of salivary proteins was demonstrated by using shotgun proteomics and two-dimensinal gel electrophoresis mass spectrometry (2-DE-MS). For the shotgun approach, whole saliva proteins were prefractionated according to molecular weight. The smallest fraction, presumably containing salivary peptides, was directly separated by capillary liquid chromatography (LC). However, the large protein fractions were digested into peptides for subsequent LC separation. Separated peptides were analyzed by on line electrospray tandem mass spectrometry (MS/MS) using a quadrupole-time of flight mass spectrometer, and the obtained spectra were automatically processed to search human protein sequence database for protein identification. Additionally, 2-DE was used to map out the proteins in whole saliva. Protein spots 105 in number were excised and in-gel digested; and the resulting peptide fragments were measured by matrix-assisted laser desorption/ionization-mass spectrometry and sequenced by LC-MS/MS for protein identification. In total, we cataloged 309 proteins from human whole saliva by using these two proteomic approaches. PMID- 15800971 TI - The wheat (Triticum aestivum L.) leaf proteome. AB - The wheat leaf proteome was mapped and partially characterized to function as a comparative template for future wheat research. In total, 404 proteins were visualized, and 277 of these were selected for analysis based on reproducibility and relative quantity. Using a combination of protein and expressed sequence tag database searching, 142 proteins were putatively identified with an identification success rate of 51%. The identified proteins were grouped according to their functional annotations with the majority (40%) being involved in energy production, primary, or secondary metabolism. Only 8% of the protein identifications lacked ascertainable functional annotation. The 51% ratio of successful identification and the 8% unclear functional annotation rate are major improvements over most previous plant proteomic studies. This clearly indicates the advancement of the plant protein and nucleic acid sequence and annotation data available in the databases, and shows the enhanced feasibility of future wheat leaf proteome research. PMID- 15800972 TI - Developmental changes in the metabolic protein profiles of wheat endosperm. AB - A combined two-dimensional gel electrophoresis-mass spectrometry approach was utilized to identify over 250 proteins of wheat (Triticum aestivum L., cv. Butte 86) starchy endosperm that participate in 13 biochemical processes: ATP interconversion reactions, carbohydrate metabolism, cell division, cytoskeleton, lipid metabolism, nitrogen metabolism, protein synthesis/assembly, protein turnover, signal transduction, protein storage, stress/defense, transcription/translation, and transport. Endosperm protein populations were compared at early (10 days post-anthesis, dpa) and late (36 dpa) stages of grain development. Analysis of protein number and spot volume revealed that carbohydrate metabolism, transcription/translation, and protein synthesis/assembly were the principal endosperm functions at 10 dpa followed by nitrogen metabolism, protein turnover, cytoskeleton, cell division, signal transduction, and lipid metabolism. Carbohydrate metabolism and protein synthesis/assembly were also major functions at 36 dpa, but stress/defense and storage were predominant. The results provide insight into biochemical events taking place during wheat grain development and highlight the value of proteomics in characterizing complex biochemical processes. Further, the proteome maps will facilitate future studies addressing the effects of genetic and environmental factors on the development and quality of wheat grain. PMID- 15800973 TI - Proteomic identification of human neutrophil alpha-defensins in chronic lung allograft rejection. AB - Chronic allograft rejection remains a leading cause of morbidity and mortality in lung transplant recipients. Currently, diagnosis is based on lung biopsies or the presence of bronchiolitis obliterans syndrome (BOS). To identify a biomarker of rejection we performed a proteome survey of archived bronchoalveolar lavage fluid (BALF) acquired from lung transplant recipients between 1993 and 1996 using mass spectrometry (MS). A total of 126 BALF samples from 57 individuals were tested. Initial MS assessment revealed numerous differences in a majority of individuals who experienced BOS, but three unusually intense peaks at m/z = 3373, 3444, and 3488. These were identified as human neutrophil peptides 1-3 (HNP). Quantification by enzyme-linked immunoabsorbent assay showed an elevated HNP level (>0.3 ng/microg protein) in 89% of patients who developed BOS2-3 within 15 months, reaching as high as 6% of the total BALF protein. In control patients, 35% demonstrated a slightly elevated HNP level that declined in all who had subsequent BALF available for testing. HNP levels did not correlate with episodes of acute rejection, cytomegalovirus or fungal infection. In conclusion, elevated HNP levels are associated with the onset of BOS and can predate the clinical onset of disease up to 15 months. PMID- 15800974 TI - Use of two-dimensional gel electrophoresis to differentiate morphospecies of Alexandrium minutum, a paralytic shellfish poisoning toxin-producing dinoflagellate of harmful algal blooms. AB - Contamination of shellfish with paralytic shellfish poisoning toxins (PST) produced by toxic harmful algal blooms (HABs) have been negatively affecting the shellfish and aquaculture industries worldwide. Therefore, accurate and early identification of toxic phytoplankton species is crucial in HABs surveillance programs that allow fish-farmers to take appropriate preventive measures in shellfish harvesting and other aquaculture activities to overcome the negative impacts of HABs on human health. The identification of toxic dinoflagellates present in the water is currently a time-consuming operation since it requires skillful taxonomists and toxicologists equipped with optical and scanning electron microscopes as well as sophisticated equipment, for example, high performance liquid chromotography-fluorescence detection. In this paper, a two dimensional gel electrophoresis (2-DE)-based proteomic approach was applied to discriminate between toxic and nontoxic strains of Alexandrium minutum. Variation in morphological features between toxic and nontoxic strains was minimal and not significant. Also, variation in 2-DE protein patterns within either toxic or nontoxic strains was low, but pronounced differences were detected between toxic and nontoxic strains. The most notable differences between these strains were several abundant proteins with pIs ranging from 4.8 to 5.3 and apparent molecular masses between 17.5 and 21.5 kDa. Groups of proteins, namely NT1, NT2, NT3, and NT4, were consistently found in all nontoxic strains, while T1 and T2 were prominent in the toxic strains. These specific protein spots characteristic for toxic and nontoxic strains remained clearly distinguishable irrespective of the various growth conditions tested. Therefore, they have the potential to serve as "taxonomic markers" to distinguish toxic and nontoxic strains within A. minutum. Initial studies revealed that the expression pattern of T1 was tightly correlated to toxin biosynthesis in the examined alga and may be used to serve as a potential toxin indicator. PMID- 15800975 TI - Overexpression of alpha enolase in hepatitis C virus-related hepatocellular carcinoma: association with tumor progression as determined by proteomic analysis. AB - To identify proteins that could be molecular targets for diagnosis and treatment of hepatitis C virus-related hepatocellular carcinoma (HCV-related HCC), we used a proteomic approach to analyze protein expression in samples of human liver. Twenty-six pairs of tumorous and corresponding nontumorous liver samples from patients with HCV-related HCC and six normal liver samples were analyzed by two dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry. One of the numerous spots that showed stronger intensity in tumorous than in nontumorous samples was identified as alpha enolase, a key enzyme in the glycolytic pathway. Expression of this protein increased with tumor dedifferentiation and was significantly higher in poorly differentiated HCC than in well-differentiated HCC. This pattern was reproduced by immunoblot analysis and immunohistochemistry. Expression of alpha enolase also correlated positively with tumor size and venous invasion. These results suggest that alpha enolase is one of the candidates for biomarkers for tumor progression that deserves further investigation in HCV-related HCC. PMID- 15800976 TI - Role of insulin/insulin-like growth factor 1 signaling pathway in longevity. AB - The insulin/insulin-like growth factor 1 (IGF-1) signaling pathway is evolutionary conserved in diverse species including C.elegans, saccharomyces cerevisiae, Drosophila melanogaster, rodents and humans, which is involved in many interrelated functions that are necessary for metabolism, growth and reproduction. Interestingly, more and more research has revealed that insulin/IGF 1 signaling pathway plays a pivotal role in the regulation of longevity. Generally, disruption of the power of this pathway will extend longevity in species ranging from C.elegans to humans. The role of insulin/IGF-1 in longevity is probably related to stress resistance. Although the underlying mechanisms of longevity are not fully understood, the Insulin/IGF-1 signaling pathway has attracted substantial attention and it will be a novel target to prevent or postpone age-related diseases and extend life span. In this review, we mainly focus on the similar constitution and role of insulin/IGF-1 signaling pathway in C.elegans, saccharomyces cerevisiae, rodents and humans. PMID- 15800977 TI - Tumor cyclooxygenase-2 levels correlate with tumor invasiveness in human hepatocellular carcinoma. AB - AIM: Recent studies suggested that cyclooxygenase-2 (COX-2) enhances tumor angiogenesis via upregulation of vascular endothelial growth factor (VEGF). Although COX-2 expression has been demonstrated in hepatocellular carcinoma (HCC), the significance of COX-2 in progression of HCC remains unclear. This study evaluated the clinico-pathological correlation of COX-2 level and its relationship with VEGF level in HCC. METHODS: Fresh tumor tissues were obtained from 100 patients who underwent resection of HCC. COX-2 protein expression was examined by immunohistochemistry, and quantitatively by an enzyme immunometric assay (EIA) of tumor cytosolic COX-2 levels. Tumor cytosolic VEGF levels were measured by an ELISA. RESULTS: Immunostaining showed expression of COX-2 in tumor cells. Tumor cytosolic COX-2 levels correlated with VEGF levels (r = 0.469, P<0.001). Correlation with clinicopathological features showed significantly higher tumor cytosolic COX-2 levels in the presence of multiple tumors (P = 0.027), venous invasion (P = 0.030), microsatellite lesions (P = 0.037) and advanced tumor stage (P = 0.008). Higher tumor cytosolic COX-2 levels were associated with worse patient survival. CONCLUSION: This study shows that elevated tumor COX-2 levels correlate with elevated VEGF levels and invasiveness in HCC, suggesting that COX-2 plays a significant role in the progression of HCC. PMID- 15800978 TI - Inhibitory effect of tumor suppressor p33(ING1b) and its synergy with p53 gene in hepatocellular carcinoma. AB - AIM: To investigate the inhibitory effect of tumor suppressor p33(ING1b) and its synergy with p53 gene in hepatocellular carcinoma (HCC). METHODS: Recombinant sense and antisense p33(ING1b) plasmids were transfected into hepatoma cell line HepG2 with lipofectamine. Apoptosis, G0/G1 arrest, cell growth rate and cloning efficiency in soft agar of HepG2 were analyzed after transfection. In three hepatoma cell lines with different endogenous p53 gene expressions, the synergistic effect of p33(ING1b) with p53 was analyzed by flow cytometry and luciferase assay was performed to detect the activation of p53 downstream gene p21(WAF1/CIP1). In addition, the expression and mutation rates of p33(ING1b) in HCC tissues were measured by immunohistochemistry and polymerase chain reaction single strand conformation polymorphism (PCR-SSCP). RESULTS: Overexpression of p33(ING1b) inhibited cell growth of HepG2, induced more apoptosis and protected cells from growth in soft agar. Combined transfer of p33(ING1b) and p53 gene promoted hepatoma cell apoptosis, G0/G1 arrest and elevated expression of p21(WAF1/CIP1). Immunostaining results showed co-localized P33(ING1b) with P53 protein in HCC tissues and there was a significant relation between protein expression rates of these two genes (P<0.01). Among 28 HCC samples, p33(ING1b) presented a low gene mutation rate (7.1%). CONCLUSION: p33(ING1b) collaborates with p53 in cell growth inhibition, cell cycle arrest and apoptosis in HCC. Loss or inactivation of p33(ING1b) normal function may be an important mechanism for the development of HCC retaining wild-type p53. PMID- 15800979 TI - Cell proliferation, apoptosis and the related regulators p27, p53 expression in hepatocellular carcinoma. AB - AIM: To investigate the expression of cell apoptosis, proliferation and the related regulators p27, p53 in hepatocellular carcinoma (HCC). METHODS: The expression of p27, p53, proliferating cell nuclear antigen (PCNA) and apoptosis in 47 HCC specimens and 42 surrounding non-cancerous tissues were detected by the immunohistochemistry and terminal deoxy-nucleotidyl transferase-mediated nick end labeling (TUNEL) technique. Meanwhile, the clinical significance of them was analyzed combining with the clinicopathological factors and follow-up data. RESULTS: (1) The average proliferating index and apoptotic index in HCC were significantly higher than that in adjacent liver tissues. The proliferating index was associated with extrahepatic metastasis. The apoptotic index was significantly lower in TNM stage I-II than in stage III-IV. The proliferating index of groups with p53-/p27+ was significantly lower than that in group with p53+/p27- (P = 0.030); (2) The level of p27 in the cytoplasmic fraction was higher in non-tumoral liver tissues and was associated with clinical stage; (3) Survival analysis showed advanced stage (P = 0.031) and with extrahepatic metastasis (P = 0.045) was significantly associated with shorter survival. In addition, the prognosis of patients with p53-/p27+ was longer than that of patients with p53+/p27- (P = 0.0356). CONCLUSION: The p53 mutation and decreased p27 expression might be involved in the imbalance of proliferation and apoptosis in HCC. Cytoplasmic displacement might lead to the inactivation of p27 protein in HCC cells and acts early during carcinogenesis of HCC. The combined examination of p27, and p53 expression allows reliable estimation of prognosis for patients with primary hepatic carcinoma. PMID- 15800980 TI - Avidin chase reduces side effects of radioimmunotherapy in nude mice bearing human colon carcinoma. AB - AIM: To evaluate the influence of avidin chase on the side effects of radioimmunotherapy (RIT) in nude mice bearing human colon carcinoma and therapeutic outcome. METHODS: Purified anti-CEA monoclonal antibody (McAb) was biotinylated with NHS-biotin, and then radiolabeled with 188Re by the direct method. 188Re-labeled biotinylated anti-CEA McAb (188Re-CEA McAb-Bt) was intravenously injected followed by intravenous injection of avidin after 24 h. SPECT imaging and biodistribution study were performed at 28-48 h after the injection of 188Re-CEA McAb-Bt. Three groups of nude mice subcutaneously grafted with human colon carcinoma were treated 7 d after the graft. Mice in the avidin chase group received intravenous injection of 188Re-CEA McAb-Bt (11.1 MBq/20 microg) followed by intravenous injection of cold avidin (80 microg) after 24 h. Mice in the control group (treated group without avidin chase) only received the injection of 188Re-CEA McAb-Bt (11.1 MBq/20 microg), another control group (non treated group) only received 0.1 mL normal saline solution. Toxicity was evaluated on the basis of change of body weight and peripheral WBC counts, and therapy effects were determined by variation in tumor volume. Histological analysis of tumors was also performed. RESULTS: Avidin chase markedly accelerated the clearance of 188Re-CEA McAb-Bt from the blood and normal tissues. The tumor uptakes of 188Re-CEA McAb-Bt at 28 h were 5.90 and 6.42% ID/g, respectively, in chase group and in non-chase group, while the tumor-to-background (T/NT) ratios were 3.19 and 0.56, respectively. The tumor uptake was slightly decreased by avidin chase, but the T/NT ratios were increased. In treated groups the growth rate of body weight and the number of WBC decreased after injection of 188Re-CEA McAb-Bt, and the WBC counts recovered earlier in the group with avidin chase than in the group without avidin chase. Compared to the non-treated group, treated groups with and without avidin chase showed significant anti-tumor effects. CONCLUSION: Avidin chase can effectively reduce the side effects of RIT, and improve therapeutic efficacy. PMID- 15800981 TI - Relationship between serum b2-microglobulin levels and virological breakthrough in HBeAg-negative chronic hepatitis B patients, under long-term treatment schedules including lamivudine. AB - AIM: Predictive value of serum b2-microglobulin (b2m) levels for virological breakthrough (VB) in HBeAg-negative chronic hepatitis B (CHB) patients under long term treatment schedules including lamivudine (LAM). METHODS: Serum b2m levels were calculated during treatment in 25 CHB patients under long-term LAM monotherapy (group A) and 12 patients under initial interferon plus LAM treatment followed by LAM monotherapy (group B), using the MEIA technology. We used Cox proportional hazard models in order to investigate the association between serum b2m levels and VB. RESULTS: Seven of 25 patients (28%), 9/25 (36%) and 14/25 (56%) from group A and 0/12, 2/12 (16.6%) and 3/12 (25%) from group B exhibited VB at months 12, 24 and 36 of treatment, respectively. All patients, from both groups, who did not show VB exhibited b2m elevation in mo 3. The duration of b2m elevation was significantly longer in the virological responder's subgroup from group A than the non-responder's one (7.3+/-2.6 vs 3.8+/-3.4 mo, P = 0.02). In comparison to group A patients whose b2m levels were increased at 3 mo, patients whose b2m levels were decreased had 4.6 times higher risk of experiencing VB (RR = 4.6, P = 0.024). When baseline variables were simultaneously included in the same Cox model, decreased b2m status was still associated with increased risk of VB (RR = 12.2, P = 0.03). CONCLUSION: In HBeAg-negative CHB patients under either long-term LAM monotherapy or initial combination treatment, serum b2m levels at 3 mo of treatment, compared to baseline ones, might be a predictor of risk for VB. PMID- 15800982 TI - Transforming growth factor-beta1 gene polymorphisms are associated with progression of liver fibrosis in Caucasians with chronic hepatitis C infection. AB - AIM: Considerable attention is focused on polymorphisms in the gene encoding transforming growth factor-beta1 (TGF-beta1), a multifunctional cytokine that is in turn a potent growth inhibitor involved in wound healing and differentiation. In humans, it promotes the pathogenesis of organ fibrosis, atherosclerosis, cancer, autoimmune and inflammatory diseases, keloid disease, and hypertrophic scarring. For this reason, much emphasis has been placed on studies elucidating the impact of TGF-beta1 and its gene variations for the susceptibility and pathogenesis of these diseases. Unfortunately, some studies have serious limitations. METHODS: We have recently described a high-throughput method for investigation the Arg25Pro polymorphism of human TGF-beta1 gene and showed that the frequency of the Pro25 allele is significantly associated with hepatic fibrogenesis. In this report, we describe two novel LightCycler (LC) techniques that facilitate the examination of the two other known alterations in the coding region of TGF-beta1. We investigated whether these polymorphisms contribute to hepatitis-induced progression of fibrogenesis in Chinese and Caucasians. RESULTS: In the Chinese ancestry, the gene polymorphisms at codons 25 and 263 were not found and the genetic variant at codon 10 is unlikely to confer susceptibility to hepatic fibrosis. Contrarily, in Caucasians TGF-beta1 allelic variations are more frequent and the presence of prolines either in codon 25 or 10 is associated with the interindividual variability in developing more severe fibrosis during chronic hepatitis C infection. CONCLUSION: In summary, these results confirm the hypothesis that TGF-beta1 polymorphisms are associated with fibrosis progression in Caucasians chronically infected with hepatitis C. PMID- 15800983 TI - Gene expression profiling reveals sequential changes in gastric tubular adenoma and carcinoma in situ. AB - AIM: To analyze the expression profiles of premalignant and/or preclinical lesions of gastric cancers. METHODS: We analyzed the expression profiles of normal gastric pit, tubular adenoma and carcinoma in situ using microdissected cells from routine gastric biopsies. For the DNA microarray analysis of formalin fixed samples, we developed a simple and reproducible RNA extraction and linear amplification procedure applying two polymerase-binding sites. The amplification procedure took only 8 h and yielded comparable DNA microarray data between formalin-fixed tissues and unfixed controls. RESULTS: In comparison with normal pit, adenoma/carcinoma showed 504 up-regulated and 29 down-regulated genes at the expected false significance rate 0.15%. The differential expression between adenoma and carcinoma in situ was subtle: 50 and 22 genes were up-, and down regulated in carcinomas at the expected false significance rate of 0.61%, respectively. Differentially expressed genes were grouped according to patterns of the sequential changes for the 'tendency analysis' in the gastric mucosa adenoma-carcinoma sequence. CONCLUSION: Groups of genes are shown to reflect the sequential expression changes in the early carcinogenic steps of stomach cancer. It is suggested that molecular carcinogenic pathways could be analyzed using routinely processed biopsies. PMID- 15800984 TI - Macrophage migration inhibitory factor stimulated by Helicobacter pylori increases proliferation of gastric epithelial cells. AB - AIM: Helicobacter pylori (H pylori) is associated with increased gastric inflammatory and epithelial expression of macrophage migration inhibitory factor (MIF) and gastric epithelial cell proliferation. This study aimed at determining whether H pylori directly stimulates release of MIF in monocytes, whether the cag pathogenicity island (PAI) is involved for this function, and whether MIF stimulated by H pylori increases gastric epithelial cell proliferation in vitro. METHODS: A cytotoxic wild-type H pylori strain (TN2), its three isogenic mutants (TN2Deltacag, TN2DeltacagA and TN2DeltacagE) were co-cultured with cells of a human monocyte cell line, THP-1, for 24 h at different organism/cell ratios. MIF in the supernatants was measured by an ELISA. Cells of a human gastric cancer cell line, MKN45, were then co-cultured with the supernatants, with and without monoclonal anti-MIF antibody for 24 h. The cells were further incubated for 12 h after addition of 3H-thymidine, and the levels of incorporation of 3H-thymidine were measured with a liquid scintillation counter. RESULTS: The wild-type strain and the isogenic mutants, TN2DeltacagA and TN2 DeltacagE, increased MIF release at organism/cell ratios of 200/1 and 400/1, but not at the ratios of 50/1 and 100/1. However, the mutant TN2delta cag did not increase the release of MIF at any of the four ratios. 3H-thymidine readings for MKN-45 cells were significantly increased with supernatants derived from the wild-type strain and the mutants TN2DeltacagA and TN2DeltacagE, but not from the mutant TN2Deltacag. Moreover, in the presence of monoclonal anti-MIF antibody, the stimulatory effects of the wild type strain on cell proliferation disappeared. CONCLUSION: H pylori stimulates MIF release in monocytes, likely through its cag PAI, but not related to cagA or cagE. H pylori-stimulated monocyte culture supernatant increases gastric cell proliferation, which is blocked by anti-MIF antibody, suggesting that MIF plays an important role in H pylori-induced gastric epithelial cell proliferation. PMID- 15800985 TI - Protective effect of low dose of melatonin against cholestatic oxidative stress after common bile duct ligation in rats. AB - AIM: To investigate the role of oxidative injury and the effect of exogenous melatonin administration on liver damage induced by bile duct ligation (BDL), and second, to evaluate the role of nitric oxide (NO), a free oxygen radical, in oxidative injury. METHODS: Thirty-two Sprague-Dawley rats were assigned to four groups: sham operation (SO), BDL, BDL+melatonin, and BDL+vehicle. Cholestasis was achieved by double ligature of the common bile duct. Melatonin was injected intraperitoneally 500 microg/(kg.d) for 8 d. Hepatic oxidative stress markers were evaluated by changes in the amount of lipid peroxides, measured as malondialdehyde (MDA), and reduced GSH. Total nitrite (NOX) concentrations were determined in hepatic homogenates. Histopathological examination was performed using a histological scoring system. RESULTS: The histopathological changes including portal inflammation, necrosis, apoptosis, focal inflammation and fibrosis were severe in the BDL and BDL+vehicle groups. There were numerous large areas of coagulation necrosis. Histological Activity Index scores of these groups were significantly higher than that of the SO group. Treatment with melatonin reduced these alterations significantly. The degree of necro-inflammation and fibrosis showed significant difference between the BDL and BDL+melatonin groups. BDL was accompanied by a significant increase in MDA and NOX, and a significant decrease in GSH levels. Mean+/-SE values of MDA, GSH and NOX levels of SO group were 147.47+/-6.69, 0.88+/-0.33 micromol/g and 180.70+/-6.58 nm/g, respectively. The values of BDL group were 200.14+/-21.30, 0.65+/-0.02 micromol/g, and 400.46+/ 48.89 nm/g, respectively, whereas the values of BDL+melatonin group were 115.93+/ 6.8, 0.74+/-0.02 micromol/g, and 290.38+/-32.32 nm/g, respectively. Melatonin treatment was associated with a significant recovery of MDA, GSH and NOX levels. CONCLUSION: We have concluded that oxidative stress is associated with the pathogenesis of cholestatic liver damage and NO contributes to oxidative damage. Melatonin, even at low dose, is an efficient agent in reducing negative parameters of cholestasis. PMID- 15800986 TI - Adenosine 5'triphosphate transport and accumulation during the cold preservation of rat hepatocytes in University of Wisconsin solution. AB - AIM: We used isolated hepatocytes to investigate how different concentrations of ATP in the University of Wisconsin (UW) solution affected both cellular ATP content and cell viability during the cold storage and the rewarming step. The mechanism involved in ATP transport and accumulation in hypothermia was also determined. METHODS: The cells were preserved up to 72 h in different conditions: UW solution without ATP (a-group), UW+5 mmol/L ATP (b-group), and UW+10 mmol/L ATP (c-group). The ATP content and the cell viability (LDH release) were determined during the cold storage and the rewarming step. In the groups a and c, the respiratory function of the cells at rewarming was studied. In addition, the cell volume of hepatocytes and the mechanism involved in ATP transport and accumulation were assessed. The extracellular degradation of exogenous nucleotides during transport experiments was investigated by a HPLC technique. RESULTS: After three days of cold storage a loss of cellular ATP content was observed in hepatocytes preserved either without nucleotides (a-group) or with 5 mmol/L ATP (b-group). In contrast, 10 mmol/L ATP (c-group) was able to maintain a normal ATP cellular content, with only a 6% diminution after 72 h of cold storage. The respiratory function was significantly different in hepatocytes preserved with 10 mmol/L ATP than without ATP. No significant change was detected for the three groups in cellular volume during the cold storage. We also report that the time course accumulation of (3H)-ATP by cold stored hepatocytes is a rapid process that is completed after 180 s with linear dependence on the extracellular ATP concentration (linear fitting results in a slope of 0.5624+/ 0.1179 mmol/L ATP intracell/mmol/L ATP extracell). CONCLUSION: Our results show that, during hypothermic storage in UW solution, hepatocytes are permeable to ATP by a diffusive mechanism. Also, we found that it is ATP the main extracellular nucleotide available for transport and it is not the breakdown products. PMID- 15800987 TI - Amylase level in extrahepatic bile duct in adult patients with choledochal cyst plus anomalous pancreatico-biliary ductal union. AB - AIM: To investigate the relationship between pancreatic amylase in bile duct and the clinico-pathological features in adult patients with choledochal cyst and anomalous pancreatico-biliary ductal union (APBDU). METHODS: From 39 patients who underwent surgery for choledochal cyst between March 1995 and March 2003, we selected 15 adult patients who had some symptoms and were radiologically diagnosed as APBDU, and their clinico-pathological features were subsequently evaluated retrospectively. However, we could not obtain biliary amylase in all the patients because of the surgeon's slip. Therefore, we measured the amylase level in gall bladder of 10 patients and in common bile duct of 11 patients. RESULTS: Levels of amylase in common bile duct and gall bladder ranged from 11,500 to 212,000 IU/L, and the younger the patients, the higher the biliary amylase level (r = -0.982, P<0.01). Pathologically, significant correlation was found between the size of choledochal cyst and the grade of inflammation (r = 0.798, P<0.01). And, significant correlation was found between the level of amylase in gall bladder and the grade of hyperplasia. On the other hand, there was no correlation to the age of symptomatic onset or inflammatory grade (r = 0.743, P<0.05). Level of lipase was elevated from 6,000 to 159,000 IU/L in bile duct and from 14,400 to 117,000 IU/L in the gall bladder; however, there was no significant correlation with age or clinico-pathological features. CONCLUSION: The results support the notion that amylase has a particular role in the onset of symptoms, and suggest that a large amount of biliary amylase induces early onset of symptom, thereby making early diagnosis possible. PMID- 15800988 TI - Use of a device that applies external kneading-like force on the abdomen for treatment of constipation. AB - AIM: To evaluate the efficacy of a recently developed device that applies kneading-like motion on the abdomen in improving constipation in elderly long term care patients. METHODS: Thirty constipated elderly patients were randomly selected from two nursing homes. They were instructed to use the device once daily for 20 min. Rate of bowel movements, volume and consistency of stool and the use of laxantia were all recorded during a 3-wk baseline period and for 12-wk treatment period. Colonic transit time (CTT) was measured in 13 patients by radiopaque markers during the baseline and at the end of treatment. RESULTS: Bowel movement rate (BM/week) increased from 1.4+/-0.4 BM/wk during baseline to 3.9+/-0.8 BM/wk during treatment (P<5.0x10(-7)). Stool amount that was "low" in 30 patients during baseline increased in 21 patients at the end of the study period (chi2 = 19.048-P = 1.3x10(-5)). Stool consistency, that was "hard" in 25 patients and "soft" in 5 patients during baseline, ameliorated in 23 patients at the end of the study (only 2 patients referred "hard" stool) (chi2 = 21.043-P = 4.0x10(-6)). The mean baseline CTT measured was 92.3+/-32.3 h at baseline and decreased to 49.4+/-31.3 h during the study period (P = 0.000208). No side effects were observed during the study period. CONCLUSION: External mechanical vibration of the abdomen reduced CTT and helped to relieve severe constipation in elderly constipated patients. PMID- 15800989 TI - Distribution of hepatitis B virus genotypes in patients with chronic hepatitis B in Turkey. AB - AIM: Hepatitis B virus (HBV) strains isolated worldwide has been classified into eight genomic groups deduced from genome comparisons and designated as genotypes A to H. We aimed to investigate prevalence of HBV genotypes and subtypes in Turkey. METHODS: A total of 88 chronic hepatitis B (CHB) patients from 15 hospitals throughout the country were included. Patients who were HBsAg positive in serum at least for 6 mo, who had HBV-DNA in serum and elevation of ALT levels more than two times upper limit of normal, and who had percutaneous liver biopsy within 6 mo were included. Genotyping of HBV was done by restriction fragment length polymorphism (RFLP). The patients received subcutaneous 9 MU interferon alpha 2a thrice a week for a period of 6 mo. RESULTS: Genotype D was detected in 78 of 88 (88.7%) patients, however, genotyping failed in two patients (2.3%), while no product was obtained in eight (9.0%) patients. Regarding subtypes, D2 was more prevalent (67 patients between 78% and 85.9%) followed by subtype D2+deletion (seven patients of 78 or 8.9%), subtype D1 (three patients of 78% or 3.9%) and subtype D3 (one patient of 78% or 1.3%). Thirty-three patients (37.5%) were HBeAg positive compared to 55 (62.5%) anti-HBe positive patients. The endpoint for the viral response of HBeAg positive patients was 27.2%, while it was found 52.7% in HBeAg negative patients (P<0.05). Long-term persistent viral response was 29.5% for all patients. CONCLUSION: This multi-center study indicates that the predominant genotype with CHB patients in Turkey like in other Mediterranean countries is genotype D. PMID- 15800990 TI - A prospective study assessing the efficacy of abdominal computed tomography scan without bowel preparation in diagnosing intestinal wall and luminal lesions in patients presenting to the emergency room with abdominal complaints. AB - AIM: To evaluate the positive predictive value of abdominal non-prepared computed tomography (CT) for diagnosing intestinal lumen or wall lesions in patients presenting to the emergency room (ER) with abdominal complaints. METHODS: For 1 year we prospectively evaluated all ER patients hospitalized after abdominal CT scan detected either intraluminal or intestinal wall lesions. These patients underwent colonoscopy serving as gold standard. Patients with prior abdominal pathology or CT findings of appendicitis or diverticulitis were excluded. RESULTS: Five hundred and sixty-eight abdominopelvic CT scans were performed in the ER, 96 had positive colonic findings. Sixty-two patients were excluded, 46 because of diverticulitis or appendicitis, 16 because of prior abdominal pathology. Of the remaining 34 patients, 14 did not undergo colonoscopy during hospitalization. Twenty eligible patients were included in the study. The positive predictive value of the CT scans performed in the ER was calculated to be 45% (95% CI 25-67). CONCLUSION: CT findings correlated with colonoscopic findings only in approximately half of the cases. Relying on non-prepared CT scan findings in planning patient management and colonoscopy may lead to unnecessary diagnostic work-ups. PMID- 15800991 TI - Serum chromogranin-A in hepatocellular carcinoma: diagnostic utility and limits. AB - AIM: The utility of serum alpha-fetoprotein (alpha-FP) for the detection of hepatocellular carcinoma (HCC) is questionable. High serum levels of chromogranin A (CgA) have recently been reported in HCC. Impaired hepatic, renal, and heart functions influence circulating CgA. The aim of this study was to assess sensitivity and specificity of serum CgA as a marker of HCC in patients with liver cirrhosis (LC). METHODS: Serum CgA levels were measured by RIA in 339 patients of which 54 HCC, 132 LC, 45 chronic hepatitis (CH), 27 chronic heart failure (CHF), 36 chronic renal failure (CRF), 45 chronic inflammatory bowel disease (IBD) as disease controls and in 75 healthy controls. Patients with liver disease or IBD and concomitant renal and/or heart failure were excluded. Pearson correlation, non-parametric combination test and confidence interval analysis were used for statistical analysis. RESULTS: Serum CgA above normal values (100 ng/mL) were found in 83% of HCC patients, in 48% of LC patients, in 20% of CH patients, in 33% of IBD patients, in 92% of CRF patients, in 100% of CHF patients, and in none of the healthy controls. The mean CgA values in HCC (769+/ 1 046), in LC (249+/-369), in CH (87+/-94), in CRF (1,390+/-1,401), in CHF (577+/ 539), in IBD (146+/-287) were significantly higher than those in healthy controls (48+/-18). HCC patients had higher CgA values (P<0.01) than LC, CH, and IBD patients but did not differ from those with CRF or CHF. The 95% CI for the mean (250-1 289 ng/mL) in HCC patients was selected as a CgA range and the lower value of such range was assumed as cut-off. Sensitivity and specificity of CgA, calculated in relation to the cut-off in patients with cirrhosis and HCC, were respectively 61% (CI 48-73%) and 82% (CI 75-88%). Serum alpha-FP values were >200 ng/mL in 21% of the HCC patients and in none of the LC patients. No significant correlation was found between alpha-FP and CgA in patients with HCC and in patients with cirrhosis. CONCLUSION: When HCC is suspected and alpha-FP is normal or <200 ng/mL, CgA serum values represent a complementary diagnostic tool, unless kidney or heart failure is present. PMID- 15800992 TI - Enhancement of leukocyte adhesion after percutaneous irradiation in rats with hepatocellular carcinoma. AB - AIM: To evaluate the effects of percutaneous radiation on leukocyte-endothelium interaction (LEI) in experimental hepatocellular carcinoma (HCC). METHODS: Twelve ACI rats underwent HCC-inoculation, six of which on day 12 received low-dose external radiation and six did not. After 12 h intravital microscopy was performed. RESULTS: LEI was significantly reduced in tumor tissue. However, irradiation of liver sinusoids and tumor tissue with 6 Gy led to a significant activation of leukocyte adhesion in the tumor with a marked increase of the proinflammatory cytokine TNF-alpha. CONCLUSION: The findings indicate that the immunological tumor-endothelial barrier can be overcome by external irradiation. PMID- 15800993 TI - Hepatic gene expression profiles associated with fibrosis progression and hepatocarcinogenesis in hepatitis C patients. AB - AIM: To determine fibrosis progression and hepatocellular carcinoma (HCC), using simultaneous gene expression analysis. METHODS: Total RNA samples were extracted from liver biopsies from 19 patients with hepatitis C virus (HCV) infection and 3 patients without HCV infection. Among the 19 HCV-infected patients, 7 and 12 patients had grade F1-2 and F3-4 fibrosis, respectively. Of the 12 patients with F3-4 fibrosis, 8 had HCC. Gene expression in the liver samples was determined using an oligonucleotide microarray. The following comparisons were performed: normal livers vs HCV-infected livers; F1-2 vs F3-4; and F3-4 with HCC vs F3-4 without HCC. Genes that were differentially expressed between these groups were identified based on signal-to-noise ratios. RESULTS: In the HCV-infected livers, genes involved in immune responses were highly expressed. Expression levels of genes for plasma proteins and drug-metabolizing enzymes were decreased and those of genes involved in the cell cycle and oncogenesis were increased in the F3-4 cases as compared to the F1-2 cases. Among the F3-4 cases, genes involved in carbohydrate metabolism tended to be more highly expressed in patients with HCC than in patients without HCC. CONCLUSION: We identified genes that are associated with fibrosis progression and hepatocarcinogenesis. This information may be used to detect increased carcinogenic potential in the livers of patients with HCV infection. PMID- 15800995 TI - Traditional Chinese medicine syndromes of chronic hepatitis B with precore mutant. AB - AIM: This study aims at exploring the distribution of TCM syndromes in CHB patients with HBV pre-core mutation (1896) and the relationship between pre-core mutation and T lymphocytes subgroup, through which to provide objective data on clinical syndrome differentiation of TCM, and further to suggest the therapeutic principle and guide clinical treatment. METHODS: One hundred and forty CHB patients were evenly divided into two study groups, HBV pre-core mutant group and HBV pre-core wild-type group. Besides, 30 healthy blood donors were selected as a healthy control group. HBV-labeled compound, T lymphocytes subgroup, and HBV-DNA pre-core mutant were tested in the study groups. T lymphocytes subgroup were also tested in the control group. All the patients were both diagnosed by syndrome differentiation of TCM and western medicine. RESULTS: The most common syndrome in mutant group was damp-heat combined with blood stasis, and the most common syndrome in the wild-type group was damp-heat stasis in the middle-jiao. There were more cases of medium and severe hepatitis in mutant group than that in wild type group. The content of CD4+ lymphocytes and CD4+/CD8+ ratio were decreased gradually (healthy control group>wild-type group>mutant group). In the wild-type group, severe and medium CHB patients had considerably lower level of them than mild CHB patients. However, in the mutant group, the opposite result appeared. Meanwhile, the content of HBV-DNA in mutant group was higher than that in wild type group. CONCLUSION: Damp, heat, toxin and blood stasis were the basic pathogens of CHB, whether pre-core mutant or not. CHB with precore mutant may lead to more severe hepatitis. The decreased content of CD4+ lymphocytes and ratio of CD4+/CD8+ may be taken as one of the indices in confirming the deficiency syndrome of CHB patients with pre-core mutation. PMID- 15800994 TI - Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi. AB - AIM: To study the effect of inhibited E-cadherin expression on invasion of cancer cells. METHODS: We designed the nucleotide sequence of siRNA corresponding to 5' non-coding and coding sequence of E-cadherin. 21-nucleotide dssiRNA was synthesized by in vitro transcription with Ambion Silencer TM siRNA Construction Kit. siRNA was transfected into gastric cancer MKN45 using TransMessenger transfection Kit. RT-PCR and immunofluorescent assay were used to investigate the inhibition of the expression of mutated E-cadherin. Invasive ability of cancer cells was determined by Transwell assay. RESULTS: The synthesis of E-cadherin mRNA rather than protein expression was suppressed dramatically 7 d after interference. Decreased protein expression was observed on d 10 after interference. On d 11, invasion ability was enhanced significantly. CONCLUSION: siRNA targeted at non-coding and coding sequence of E-cadherin showed significant inhibition on mRNA and protein expression. Inhibited E-cadherin expression results in increased invasion ability of cancer cells. PMID- 15800996 TI - Stomach-interposed cholecystogastrojejunostomy: a palliative approach for periampullary carcinoma. AB - AIM: For patients of periampullary carcinoma found to be unresectable at the time of laparotomy, surgical palliation is the primary choice of treatment. Satisfactory palliation to maximize the quality of life with low morbidity and mortality is the gold standard for a good procedure. Our aim is to explore such a procedure as an alternative to the traditional ones. METHODS: A modified double bypass procedure is performed by, in addition to the usual gastrojejunostomy, implanting a mushroom catheter from the gall bladder into the jejunum through the interposed stomach as an internal drainage. A retrospective review was performed including 22 patients with incurable periampullary carcinomas who underwent this surgery. RESULTS: Both jaundice and impaired liver function improved significantly after surgery. No postoperative deaths, cholangitis, gastrojejunal, biliary anastomotic leaks, recurrent jaundice or late gastric outlet obstruction occurred. Delayed gastric emptying occurred in two patients. The total surgical time was 150+/-26 min. The estimated blood loss was 160+/-25 mL. The mean length of hospital stay after surgery was 22+/-6 d. The mean survival was 8 mo (range 1.5-18 mo). CONCLUSION: In patients of unresectable periampullary malignancies, stomach-interposed cholecystogastrojejunostomy is a safe, simple and efficient technique for palliation. PMID- 15800997 TI - Liver fibrosis caused by choledocholith to regress after biliary drainage. AB - AIM: To study the correlation between liver fibrosis severity and biliary drainage in patients with choledocholith. METHODS: A follow-up study on seven patients with liver fibrosis due to choledocholith was made. The data, including biochemical tests (aspartate aminotransferase, alanine aminotransferase) and liver histological features before and after biliary drainage, were collected and studied. The fibrosis severity was scored on a scale from 0 to 3, with 0 denoting none, 1 portal and periportal fibrosis, 2 the presence of numerous fiber septa, and 3 cirrhosis. The average liver fibrosis severity scores of the first and second biopsy were compared with statistical method. RESULTS: The first, second liver fibrosis severity scores of these seven patients were 2,1; 2,1; 1,0; 1,1; 2,1; 2,1; 1,0 respectively. The results showed that the average liver fibrosis severity score of the second liver biopsy decreased significantly compared with the first liver biopsy (n = 7, t = 4.25, P<0.05). CONCLUSION: Liver fibrosis due to choledocholith may regress after biliary drainage. PMID- 15800998 TI - Effects of psychological stress on small intestinal motility and bacteria and mucosa in mice. AB - AIM: To investigate the effects of psychological stress on small intestinal motility and bacteria and mucosa in mice, and to explore the relationship between small intestinal dysfunction and small intestinal motility and bacteria and mucosa under psychological stress. METHODS: Sixty mice were randomly divided into psychological stress group and control group. Each group were subdivided into small intestinal motility group (n = 10), bacteria group (n = 10), and D-xylose administered to stomach group (n = 10). An animal model with psychological stress was established housing the mice with a hungry cat in separate layers of a two layer cage. A semi-solid colored marker (carbon-ink) was used for monitoring small intestinal transit. The proximal small intestine was harvested under sterile condition and processed for quantitation for aerobes (Escherichia coli) and anaerobes (Lactobacilli). The quantitation of bacteria was expressed as log10(colony forming units/g). D-xylose levels in plasma were measured for estimating the damage of small intestinal mucosa. RESULTS: Small intestinal transit was inhibited (39.80+/-9.50% vs 58.79+/-11.47%, P<0.01) in mice after psychological stress, compared with the controls. Psychological stress resulted in quantitative alterations in the aerobes (E. coli). There was an increase in the number of E. coli in the proximal small intestinal flora (1.78+/-0.30 log10 (CFU/g) vs 1.37+/-0.21 log10 (CFU/g), P<0.01), and there was decrease in relative proportion of Lactobacilli and E. coli of stressed mice (0.53+/-0.63 vs 1.14+/ 1.07, P<0.05), while there was no significant difference in the anaerobes (Lactobacilli) between the two groups (2.31+/-0.70 log10 (CFU/g) vs 2.44+/-0.37 log10 (CFU/g), P>0.05). D-xylose concentrations in plasma in psychological stress mice were significantly higher than those in the control group (2.90+/-0.89 mmol/L vs 0.97+/-0.33 mmol/L, P<0.01). CONCLUSION: Small intestinal dysfunction under psychological stress may be related to the small intestinal motility disorder and dysbacteriosis and the damage of mucosa probably caused by psychological stress. PMID- 15800999 TI - Hypermethylation of CpG island in O6-methylguanine-DNA methyltransferase gene was associated with K-ras G to A mutation in colorectal tumor. AB - AIM: To investigate the functions of promoter hypermethylation of O6 methylguanine-DNA methyltransferase (MGMT) gene in colorectal tumorigenesis and progression. METHODS: The promoter hypermethylation of MGMT gene was detected in 27 sporadic colorectal adenomas, 62 sporadic colorectal carcinomas and 20 normal colorectal mucosa tissues by methylation-specific PCR. At the same time, the expression of MGMT protein was carried out in the same samples using immunohistochemistry. Mutant-allele-specific amplification was used to detect K ras G to A point mutation in codon 12. RESULTS: None of the normal colorectal mucosa tissues showed methylated bands. Promoter hypermethylation was detected in 40.7% (11 of 27) of adenomas and 43.5% (27 of 62) of carcinomas. MGMT proteins were expressed in nucleus and cytoplasm of normal colorectal mucosa tissues. Loss of MGMT expression was found in 22.2% (6 of 27) of adenomas and 45.2% (28 of 62) of carcinomas. The difference between them was significant (P = 0.041). In the 6 adenomas and 28 carcinomas losing MGMT expression, 5 and 24 cases presented methylation, respectively (P = 0.027, P<0.001). Thirteen of the 19 colorectal tumors with K-ras G to A point mutation in codon 12 had methylated MGMT (P = 0.011). The frequencies of K-ras G to A point mutation were 35.3% (12 of 34) and 12.7% (7 of 55) in tumors losing MGMT expression and with normal expression, respectively. CONCLUSION: Promoter hypermethylation and loss of expression of MGMT gene were common events in colorectal tumorigenesis, and loss of expression of MGMT occurs more frequently in carcinomas than in adenomas in sporadic patients. Hypermethylation of the CpG island of MGMT gene was associated with loss of MGMT expression and K-ras G to A point mutation in colorectal tumor. The frequency of K-ras G to A point mutation was increased in tumors losing MGMT expression. It suggests that epigenetic inactivation of MGMT plays an important role in colorectal neoplasia. PMID- 15801001 TI - An autopsy case of acute pancreatitis with a high serum IgG4 complicated by amyloidosis and rheumatoid arthritis. AB - We report an autopsy case of acute pancreatitis with a high serum IgG4 concentration complicated by systemic amyloid A amyloidosis and rheumatoid arthritis (RA). The patient was a 42-year-old Japanese female with a 22-year history of rheumatoid arthritis. She was diagnosed with myasthenia gravis when she was 31-year old. At the onset of pancreatitis, the patient was anti-nuclear antibody-positive, and had high serum gamma globulin and IgG4 levels. Dexamethasone and conventional therapy induced clinical remission and significantly decreased the serum IgG4 and gamma globulin. However, despite the decreased disease parameters, the patient developed a bleeding pseudocyst and died of cardiac failure. In the autopsy examination, it was determined that pancreatitis was probably caused by ischemia due to vascular obstruction caused by amyloid deposition in the pancreas. Even though acute pancreatitis is a rare complication in RA patients, we speculate that an autoimmune pancreatitis-related mechanism and ischemia due to vascular obstruction by amyloid deposition might be attributable to a single source that leads to acute pancreatitis in our particular case. PMID- 15801000 TI - Identification of effective siRNA against K-ras in human pancreatic cancer cell line MiaPaCa-2 by siRNA expression cassette. AB - AIM: We shall construct the small interfering RNA (siRNA) expression cassette (SEC) targeting activated K-ras gene sequence, identify more effective siRNA sequence against K-ras gene in human pancreatic cancer cell line MiaPaCa-2 by SEC and reveal the anti-cancer effects of RNA interference (RNAi) and its therapeutic possibilities. METHODS: Three different sites of SECs were constructed by PCR. K1/siRNA, K2/siRNA and K3/siRNA are located at sites 194, 491 and 327, respectively. They were transfected into MiaPaCa-2 cells by liposome to inhibit the expression of activated K-ras. In the interfering groups of sites 194 and 491, we detected the apoptosis in cells by FACS after they were incubated for 48 h, then we tested the alternation of K-ras gene in MiaPaCa-2 cells by RT-PCR immunofluorescence, respectively. RESULTS: Introduction of the K1/siRNA and K2/siRNA against K-ras into MiaPaCa-2 cells leads to increased apoptosis, and the number of apoptotic cells is increased compared with control cells. The tests of RT-PCR immunofluorescence show the effects of inhibiting expression of activated K-ras gene by RNA interference in the K1/siRNA and K2/siRNA groups. We also find that the introduction of K3/siRNA has no effect on MiaPaCa-2 cells. CONCLUSION: K1/siRNA and K2/siRNA can inhibit the expression of activated K-ras but K3/siRNA has no effect, demonstrating that K1/siRNA and K2/siRNA are effective sequences against K-ras gene and K3/siRNA are not. We conclude that specific siRNA against K-ras expression may be a powerful tool to be used therapeutically against human pancreatic cancer. PMID- 15801002 TI - Transhepatic fibrinolysis of mesenteric and portal vein thrombosis in a patient with ulcerative colitis: a case report. AB - AIM: To present a case of acute mesenteric and portal vein thrombosis treated with thrombolytic therapy in a patient with ulcerative colitis in acute phase and to review the literature on thrombolytic therapy of mesenteric-portal system. Treatment of acute portal vein thrombosis has ranged from conservative treatment with thrombolysis and anticoagulation therapy to surgical treatment with thrombectomy and/or intestinal resection. METHODS: We treated our patient with intraportal infusion of plasminogen activator and then heparin through a percutaneous transhepatic catheter. RESULTS: Thrombus resolved despite premature interruption of the thrombolytic treatment for neurological complications, which subsequently resolved. CONCLUSION: Conservative management with plasminogen activator, could be considered as a good treatment for patients with acute porto mesenteric thrombosis. PMID- 15801003 TI - Hypokalemic myopathy in a patient with gluten-sensitive enteropathy and dermatitis herpetiformis Duhring: a case report. AB - The case of a 22-year-old patient with symptomatic hypokalemia caused by rhabdomyolysis is presented as a rarely reported complication of gluten-sensitive enteropathy (GSE) and dermatitis herpetiformis Duhring. The patient's myopathy ceased on potassium supplementation and her other complaints resolved while on gluten-free diet. Recovery was otherwise uneventful with a rapid decline in serum CPK level. At the time of her last follow-up a few months later, she was free of symptoms and CPK remained stable. Patients with GSE may present with hypokalemia in association with diarrhea and emesis, and if potassium loss is rapid, rhabdomyolysis may occur. PMID- 15801004 TI - Regression of liver metastases of occult carcinoid tumor with slow release lanreotide therapy. AB - Few clinical studies have demonstrated an anti-proliferative activity of somatostatin (SST) analogs in carcinoids. We report the case of a woman with liver metastases of neuroendocrine tumor and no evidence of the primary tumor. The liver metastases were characterized by high proliferation index, immunoreactiviy for somatostatin receptor (SSTR)-1, 2, 3 and 5 and positive octreoscan. Urinary 5-hydroxyindolacetic acid, serum serotonin and chromogranin A were elevated. Slow release lanreotide (SR-LAN) therapy for 3 mo controlled clinical and biochemical signs of carcinoid tumor and caused a clear-cut reduction in the diameter of two liver metastases and disappearance of another lesion, with further reduction after 6 and 18 mo. We demonstrated a clear-cut long-lasting anti-proliferative effect of SR-LAN on liver metastases of occult carcinoid with high proliferation index and immunoreactivity for SSTR-1, 2, 3, and 5. Immuno-histochemistry for SSTRs could be a suitable method for the selection of patients with metastatic carcinoid that may benefit from SST analog therapy. PMID- 15801005 TI - Mucinous nonneoplastic cyst of the pancreas: a truly novel pathological entity? AB - Recently, a novel and distinct pancreatic cystic tumor termed "mucinous nonneoplastic" cyst was described in the literature. We report our experience with a 71-year-old female with a cystic tumor in the body of the pancreas demonstrating features suggestive of this diagnosis. We also review the literature regarding this "novel" pathological entity and discuss critically its existence and its differential diagnoses. PMID- 15801006 TI - Cardiac metastases of gallbladder carcinoma. AB - This report describes the case of a 68-year-old woman diagnosed with advanced gallbladder cancer, whose autopsy revealed multiple metastases, including cardiac metastases. PMID- 15801007 TI - Everyone has the right to work. PMID- 15801008 TI - Patient questionnaires for clinical research and improved standard patient care: is it better to have 80% of the information in 100% of patients or 100% of the information in 5% of patients? PMID- 15801009 TI - Biochemical markers for osteoarthritis: from the present to the future and back to the past. PMID- 15801010 TI - Looking out for the backside! PMID- 15801011 TI - Expanding the definition of clinical differences: from minimally clinically important differences to really important differences. Analyses in 8931 patients with rheumatoid arthritis. AB - OBJECTIVE: Minimally clinically important differences (MCID) have become an important way to interpret data of randomized clinical trials (RCT), but do not reflect the degree of improvement consistent with a "really important difference" (RID). To define RID, we compared mean and/or least desirable clinical states with best and/or most desirable states. METHODS: In total, 8931 patients with rheumatoid arthritis (RA) < 65 years of age completed the Health Assessment Questionnaire (HAQ) and Medical Outcomes Survey Short Form 36 Physical Component Score (PCS). Definitions of RID were based on values for HAQ and PCS corresponding with the best and worst category of the following conditions: disabled vs not disabled: joint replacement vs no joint replacement; < poverty level vs > poverty level; very satisfied with health vs not; and independent in participation activities vs not. RESULTS: In contrast to published MCID values for the HAQ of approximately 0.22, RID was as high as 0.76 using objective reference conditions and 0.87 using the subjective measure of dependence vs independence. The HAQ score of independent RA patients was 0.38 (SD 0.45), and was 0.42 (SD 0.53) for those very satisfied with their health. The difference in HAQ scores between disabled and working patients was approximately 0.75. PCS differences were similarly increased. CONCLUSION: RID values are 3 to 4 times greater than MCID values. Although MCID are meaningful statistics for RCT, the RID percentage achieved [(actual improvement/RID) 100%] is a simple way to put the results of RCT in a broader perspective that gives an idea of how much additional treatment effect is needed. PMID- 15801012 TI - Physical function and health related quality of life: analysis of 2-year data from randomized, controlled studies of leflunomide, sulfasalazine, or methotrexate in patients with active rheumatoid arthritis. AB - OBJECTIVE: To determine whether improvements in physical function and health related quality of life (HRQOL) are sustained over 2 years of blinded treatment with leflunomide (LEF), methotrexate (MTX), or sulfasalazine (SSZ) in patients with active rheumatoid arthritis (RA). METHODS: Three phase III randomized, controlled trials compared LEF, MTX, and SSZ in patients with active RA. Improvements in physical function were assessed by Health Assessment Questionnaire Disability Index (HAQ-DI) and Modified Health Assessment Questionnaire (MHAQ); monthly MHAQ and mean HAQ scores were used to calculate American College of Rheumatology responses; HAQ-DI was assessed at baseline and 6 month intervals. In US301, the Medical Outcomes Study 36-Item Short-Form questionnaire (SF-36) assessed treatment-associated changes in HRQOL at baseline and 6-month intervals. RESULTS: Mean and median improvements in HAQ-DI after 12 and 24 months of active treatment in all phase III protocols significantly exceeded -0.22 or a minimum clinically important difference (MCID). These improvements closely reflected positive changes in SF-36 that met or exceeded MCID in all domains with LEF and MTX treatment. Problem Elicitation Technique Top 5 scores reflected improvements in performance of physical activities most important to patients. CONCLUSION: Improvements in physical function were sustained over 24 months of successful treatment with LEF, MTX, and SSZ, and reflected improvements in mental as well as physical domains of HRQOL. PMID- 15801013 TI - A pilot study to determine whether disability and disease activity are different in African-American and Caucasian patients with rheumatoid arthritis in St. Louis, Missouri, USA. AB - OBJECTIVE: To compare the levels of disability and disease activity in African Americans and Caucasians with rheumatoid arthritis (RA) in an academic medical center practice, and to determine whether the differences are independently associated with ethnicity. METHODS: Data on socioeconomic, disease related, psychological, and behavioral variables were obtained from 100 outpatients (67 Caucasians, 33 African-Americans) with RA. Functional status was assessed with the Health Assessment Questionnaire (HAQ) and Disease Activity Score (DAS-28). Chi-square and Student t tests were used to test for differences between groups. Multiple regression analysis was used to determine whether ethnicity was associated with these differences independent of other factors known to influence disease outcome. RESULTS: African-Americans and Caucasians did not differ with respect to age, sex, disease duration, rheumatoid factor positivity, and medication compliance. African-Americans had higher scores than Caucasians for HAQ (1.5 +/- 0.8 vs 0.9 +/- 0.7; p < 0.001) and DAS-28 (5.5 +/- 1.3 vs 4.3 +/- 1.4; p < 0.001). Regression models showed that ethnicity was not independently associated with the higher HAQ and DAS-28 scores when controlled for demographic, socioeconomic, psychological, and behavioral factors. Arthritis self-efficacy approaches significance in the regression model. CONCLUSION: HAQ disability and RA disease activity were higher in African-Americans than Caucasians in this sample from an academic medical center practice. However, ethnicity was not independently associated with these outcomes when socioeconomic and psychological factors were taken into account. Improvement in self-efficacy has the potential to improve outcome in African-Americans with RA. PMID- 15801014 TI - Tuberculosis in systemic lupus erythematosus in an endemic area and the role of isoniazid prophylaxis during corticosteroid therapy. AB - OBJECTIVE: The efficacy of isoniazid (INAH) prophylaxis against tuberculosis (TB) in patients taking corticosteroid remains controversial. Hong Kong is an endemic area for TB, with an annual risk of 0.11/100 in the general population. Patients with systemic lupus erythematosus (SLE) have an increased susceptibility to TB because of their intrinsic immunocompromised state and the use of corticosteroid therapy. We examined the usefulness of INAH in the prevention of recurrences of TB in patients with SLE receiving high dose corticosteroid therapy. METHODS: Medical records of a cohort of patients with SLE were reviewed. Patients with a history of TB who had previously been adequately treated were retrospectively examined for subsequent recurrence of TB. A comparison was performed based on the use of INAH at the discretion of the attending physician in some patients (INAH group) but not others (non-INAH group) during lupus exacerbation that required the use of prednisolone >/= 15 mg/day or equivalent. RESULTS: A total of 91 episodes of TB from 76 individuals in a cohort of 652 SLE patients with a duration of followup of 13.9 +/- 7.5 years were identified (prevalence of 1.06/100 patient-years). 43 episodes were given INAH while 48 were not. There were 18 recurrences of TB (recurrence rate of 1.66/100 patient-years). Recurrence rates in the INAH and non-INAH groups were 1.59 and 1.74 per 100 patient-years (p = 0.72). However, patients in the INAH group had more lupus exacerbations. Further, extrapulmonary TB was also found to have a higher recurrence rate than pulmonary TB. A case-controlled analysis was thus performed (n = 46) matching patients for the number of lupus exacerbations, cumulative doses of prednisolone, and initial site of TB. There was no difference in the recurrence rates of TB between the matched INAH (0.55/100 patient-years) and non-INAH (1.100 patient years) groups (p = 0.66). CONCLUSION: Patients with SLE have a higher prevalence of TB infection than the general population. They are at risk of TB recurrence when given immunosuppressive doses of corticosteroid. PMID- 15801015 TI - Effects of prasterone on bone mineral density in women with systemic lupus erythematosus receiving chronic glucocorticoid therapy. AB - OBJECTIVE: To assess the effects of treatment with prasterone (dehydroepiandrosterone) on bone mineral density (BMD) in female patients with mild to moderate systemic lupus erythematosus (SLE) receiving chronic treatment with glucocorticoids. METHODS: Fifty-five female patients with SLE who had received prednisone (or glucocorticoid equivalent) /= 6 months were treated for 1 year with either prasterone 200 mg/day (n = 24) or placebo (n = 31) in this randomized, double blind trial. Prasterone or placebo was added to each patient's one or more concomitant standard SLE medications, including glucocorticoids, nonsteroidal antiinflammatory drugs, antimalarials, methotrexate, azathioprine, and other immunosuppressives, which were to be maintained at fixed doses for the duration of the study. RESULTS: BMD was significantly improved in patients who received prasterone compared to placebo. At the lumbar spine, there was a mean (SEM) gain in BMD of 1.7 +/- 0.8% in the prasterone group compared to a mean loss in BMD of -1.1 +/- 0.5% in the placebo group (p = 0.003 between groups). For the total hip, mean gain was 2.0 +/- 0.9% in the prasterone group vs a mean loss of -0.3 +/- 0.4% in the placebo group (p = 0.013 between groups). In the prasterone treatment group, the mean gains from baseline at both lumbar spine and hip were statistically significant. CONCLUSION: Prasterone treatment prevented BMD loss and significantly increased BMD at both the lumbar spine and total hip in female patients with SLE receiving exogenous glucocorticoids. PMID- 15801016 TI - Antiphospholipid antibody profiles and their clinical associations in Chinese patients with systemic lupus erythematosus. AB - OBJECTIVE: Different prevalences of antiphospholipid antibodies (aPL) have been reported in different populations of patients with systemic lupus erythematosus (SLE). Chinese are generally believed to have lower risk of vascular thrombosis. We examined the prevalence of aPL including lupus anticoagulant (LAC), anticardiolipin (aCL) and anti-beta2-glycoprotein I (anti-beta2-GPI) antibodies, the level of thrombotic risk, and the association of aPL with thrombotic and pregnancy outcomes in a Chinese cohort with SLE at the university lupus clinic during the period 1986-2003. METHODS: aPL were measured in 272 SLE patients, and medical records were reviewed for vascular thrombosis and pregnancy outcomes. RESULTS: The prevalence of LAC, IgG aCL, and IgG anti-beta2-GPI antibodies was 22.4%, 29.0%, and 7.7%, respectively. There were 38 episodes of thrombosis after a mean duration of followup of 11.0 +/- 6.8 SD years, giving a thrombotic rate of 1.26/100 patient-years. All aPL were shown to be associated with vascular thrombosis. IgG anti-beta2-GPI antibodies were found to be associated with recurrent thrombosis [8.0/100 patient-years or 25.0% (7/28)]. Patients taking hydroxychloroquine were found to have fewer thrombotic complications than those who were not (OR 0.17, 95% CI 0.07-0.44; p < 0.0001). LAC was the strongest factor associated with recurrent miscarriages [relative risk 12.3, 95% CI 1.22 123.31; p = 0.03). The diagnosis of secondary antiphospholipid syndrome was satisfied in 8.9% of patients. CONCLUSION: The lifetime and recurrent thrombotic rates in our patients with aPL were not particularly different from those in the literature. However, the lower prevalence of aPL in our cohort may suggest a role of other prothrombotic factors in predisposition to thrombosis. PMID- 15801017 TI - Anti-lipoprotein lipase antibody in systemic sclerosis: association with elevated serum triglyceride concentrations. AB - OBJECTIVE: The vascular damage systemic sclerosis (SSc) consists mainly of microvascular changes, but recently macrovascular changes with dyslipidemia were recognized. In systemic lupus erythematosus (SLE), autoantibody to lipoprotein lipase (LPL), a key enzyme that hydrolyzes triglycerides, suggested a role of autoimmunity for elevated serum triglyceride levels and atherosclerosis. We investigated the prevalence and levels of anti-LPL antibodies, their clinical correlation, and their functional significance in patients with SSc. METHODS: Serum samples from patients with diffuse cutaneous SSc (dSSc; n = 55), limited cutaneous SSc (lSSc; n = 75), SLE (n = 21), and dermatomyositis (DM; n = 21) and healthy controls (n = 41) were examined by ELISA. The presence of anti-LPL antibody was evaluated by immunoblotting analysis using purified LPL. To determine the functional relevance of anti-LPL antibody in vivo, we assessed whether anti-LPL autoantibody was able to inhibit LPL activity using the LPL activity kit. RESULTS: ELISA revealed that IgG or IgM anti-LPL antibodies were detected in 35% of SSc patients, while they were also positive in 67% of SLE patients and 43% of DM patients. The presence of IgG anti-LPL antibody was associated with elevated serum triglyceride levels, greater extent of skin fibrosis, and more frequent presence of lung fibrosis, heart involvement, and anti-topoisomerase I antibodies. The presence of anti-LPL autoantibody was confirmed by immunoblotting analysis. LPL activity was inhibited by IgG anti-LPL antibodies in sera from SSc patients with elevated serum triglyceride levels. CONCLUSION: Our results suggest that anti-LPL autoantibody contributes to elevated serum triglyceride levels by inhibiting LPL enzyme activity in patients with SSc. PMID- 15801018 TI - Increased incidence of carcinoma of the tongue in patients with systemic sclerosis. AB - OBJECTIVE: To describe the incidence of carcinoma of the tongue in a large cohort of patients with systemic sclerosis (SSc). METHODS: In total, 769 patients with SSc were prospectively followed over 16 years for the development of cancer. Patients with a diagnosis of carcinoma of the tongue were identified to determine the incidence of this cancer in SSc. The results were compared to the incidence of tongue cancer in the SEER cancer registries. RESULTS: A total of 3775 patient years of followup of 769 patients with SSc (392 diffuse cutaneous, 377 limited cutaneous) prospectively evaluated for the occurrence of cancer disclosed 9 patients who were diagnosed with oral cavity and pharyngeal carcinomas. Six of these patients had squamous cell carcinoma of the tongue. One of these had both pharyngeal and tongue squamous cell carcinomas within a 4-year period, and another had 3 separate squamous cell carcinomas of the tongue. The standardized incidence ratio of squamous cell carcinoma of the tongue observed in this cohort of patients with SSc was 25-fold higher than that expected in an age adjusted population from the SEER cancer registries. All patients with SSc identified within this cohort with oral cavity carcinomas had the diffuse subset of the disease. CONCLUSION: There is a highly significant increase in the incidence of squamous cell carcinoma of the tongue in patients with SSc. A remarkable observation was that all patients within this cohort who developed oral cancer had the diffuse subset of SSc. This suggests a relationship between the etiology or pathogenesis of the diffuse form of SSc and development of squamous cell carcinoma of the tongue in this group of patients. PMID- 15801019 TI - Surgery of the hand in patients with systemic sclerosis: outcomes and considerations. AB - OBJECTIVE: To assess the current status of hand surgery in patients who have systemic sclerosis (SSc) and to elucidate special issues of surgery in this patient group. METHODS: A systematic review of English language original studies of surgical procedures of the hand in patients with SSc was performed using Medline, PreMedline, Embase, and Web of Science, from 1975 to March 15, 2004. RESULTS: Thirty-four studies were reviewed: 5 describing surgical procedures on joints, 13 on calcinosis removal, and 20 on digital sympathectomy. When the hand is affected by advanced contracture and deformity due to scleroderma, a nominal measured improvement in position and function may lead to a substantial improvement in the patient's adaptive ability to perform certain activities of daily living. A major concern is the potential for postoperative digital ischemia secondary to vascular involvement, as most of these patients exhibit blood vessel wall changes and Raynaud's phenomenon. Surgical wounds generally heal well following fusion of the proximal interphalangeal (PIP) or distal interphalangeal joint. Correction of severe flexion contractures of the PIP joint improves function and may reduce the frequency of dorsal skin ulceration. Recurrent digital tip ulceration occurs in 31.8-71.4% (median 45.2%) of scleroderma patients, reported to progress to gangrene and autoamputation in 14-29% of cases. Microsurgical revascularization of the hand, digital arterial reconstruction, and peripheral sympathectomy may improve digital vascular perfusion, heal digital ulcers, and relieve pain. Subcutaneous calcifications occur in 8.9-73.1% (median 44.1%) of SSc patients, most commonly at the fingertip, causing pain, functional impairment, and ulceration. Calcinosis can be partially removed with a high-speed burr or carbon dioxide laser. CONCLUSION: The goals of surgery for advanced SSc affecting the hand are limited and include pain relief through sympathectomy and increased perfusion, repositioning the digit, providing a functional position of fusion, and modest mobilization through resection arthroplasty. PMID- 15801020 TI - Kidney disease other than renal crisis in patients with diffuse scleroderma. AB - OBJECTIVE: To determine the frequency and severity of kidney abnormalities in patients with diffuse scleroderma. METHODS: All patients with diffuse scleroderma seen at the University of Pittsburgh between 1972 and 1993 were included in the study. Kidney function tests were routinely obtained as part of the Pittsburgh Scleroderma Outcome Study. Additional kidney tests were obtained as part of the 1992 biannual outcome assessment. Patients who had kidney abnormalities including a serum creatinine > 1.2 mg/dl or proteinuria prior to 1993 were identified. The clinical setting and longterm outcome of kidney disease were evaluated. RESULTS: Renal crisis occurred in 129/675 (19.5%) patients. Kidney function abnormalities or proteinuria were present in 173 (26%); 48% had no abnormalities. Most patients had other explanations for the abnormality. Only 12 (2%) of the 675 patients with diffuse scleroderma had no explanation for the elevated creatinine level. Most patients with proteinuria had toxicity from D-penicillamine. No explanations for proteinuria were found in 16 (2%) of this cohort. Thus, a total of only 28 (4%) of these 675 patients had an unknown cause for their kidney dysfunction or proteinuria. None of these patients, who were followed for a mean of 10 years after onset of scleroderma, have developed chronic renal insufficiency that progressed to dialysis. CONCLUSION: Patients with diffuse scleroderma without renal crisis rarely have significant increases in serum creatinine or proteinuria that cannot be explained by other etiologies. These patients with scleroderma should be carefully evaluated for non-scleroderma causes of kidney disease. PMID- 15801021 TI - Effect of granulocyte macrophage-colony stimulating factor on extracellular matrix deposition by dermal fibroblasts from patients with scleroderma. AB - OBJECTIVE: . To investigate the in vitro effect of granulocyte macrophage-colony stimulating factor (GM-CSF) on the deposition of extracellular matrix (ECM) in fibroblasts obtained from the skin of patients with systemic sclerosis (Ssc), compared to healthy controls. METHODS: Dermal fibroblasts obtained from 14 patients with SSc (7 with the diffuse form and 7 with CREST syndrome) and from 7 controls were studied. Both SSc and normal skin fibroblast cultures were stimulated for 4 and 8 days with 100 ng/ml GM-CFS. GM-CSF receptor (GM-CSFR) expression was determined by Western blot of cell lysates. Immunofluorescence was used to determine GM-CSFR expression and to investigate the deposition of ECM (type I collagen, fibronectin, and tenascin). Quantitative analysis of ECM was performed by ELISA. Expression of type I collagen and metalloproteinase 1 (MMP-1) mRNA was determined by real-time quantitative PCR. RESULTS: Deposition of ECM by normal fibroblasts appeared not to be influenced by stimulation with GM-CSF; in contrast, after stimulation with GM-CSF SSc fibroblasts showed increased deposition of fibronectin and tenascin, while type I collagen production was decreased; these results were found with both immunofluorescence and ELISA. Quantitative PCR revealed that GM-CSF inhibited the expression of mRNA type I collagen in SSc fibroblasts but not in normal fibroblasts, whereas levels of the main collagenolytic enzyme, MMP-1, were not affected. CONCLUSION: These results suggest that in SSc fibroblasts GM-CSF exerts a blocking effect on the deposition of type I collagen, through an inhibitory action on mRNA, while the production of other components of ECM such as fibronectin and tenascin is increased by stimulation with this cytokine. PMID- 15801022 TI - Perinatal characteristics and risk of developing primary Sjogren's syndrome: a case-control study. AB - OBJECTIVE: To study perinatal characteristics as risk factors for developing primary Sjogren's syndrome (SS). METHODS: This was a case control study with extraction of information from birth records comprising 32 cases with SS (fulfilling the unified American-European classification criteria) and 159 controls. Cases were selected from a patient register of SS cases in Malmo, Sweden. For each case, 5 controls (living in the same catchment area, matched by date of birth, sex, and delivery unit) from the general population were identified. The relative risks of developing SS were assessed as odds ratios (OR). The primary predictor searched for was birth weight. Secondary predictors were breastfeeding during postpartum hospital stay, paternal occupation, placenta weight, gestational length, diseases during pregnancy, maternal age, parity, and history of miscarriage. RESULTS: Significantly increased OR were observed for high birth weight (>/= 4000 vs 3000-3999 g, OR = 3.8 95% confidence interval, CI: 1.3-11.7) and low maternal age (p < 0.05). Low paternal socioeconomic status (OR = 3.2, 95% CI: 1.0-10.5) and being first-born (OR = 2.5 95% CI: 1.0-5.0) tended to be associated with SS. CONCLUSIONS: Our findings suggest that characteristics of the perinatal period may be of etiologic importance in the pathogenesis of SS. Possible mechanisms include modulation of the immune system early in life. It is conceivable that birth weight may be a marker for qualitative and/or quantitative differences in the immune system. PMID- 15801023 TI - Computer-administered bath ankylosing spondylitis and Quebec Scale outcome questionnaires for low back pain: agreement with traditional paper format. AB - OBJECTIVE: To measure the agreement between computer and paper-administered versions of Bath ankylosing spondylitis (AS) questionnaires and the Quebec Scale for low back pain (LBP). METHODS: Fifty patients with LBP completed the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Functional Index (BASFI), Global Score (BAS-G), and the Quebec Scale. Outcome measures were administered both in traditional paper format and by computerized touch-screen system. The order of completion was randomly assigned to each participant. The length of time required to complete each set of questionnaires was recorded and a "washout" period of at least 40 minutes was ensured between completion of the first and second set of outcome measures. RESULTS: There was no statistically significant difference in completion time between the 2 methods of administration. A small systematic difference between computer and paper-administered versions was observed in the Quebec Scale and in the BAS-G results. However, there was a high degree of agreement between paper and computer-administered versions of the Quebec Scale, the BASDAI, BASFI, and BAS-G. Out of the 50 subjects, 84% indicated a preference for the computer-administered method. CONCLUSION: The Bath AS questionnaires and the Quebec Scale can be reliably administered by a computerized touch-screen system. Given the ease of data integration and analysis supported by computer-administered versions of these outcome measures, their excellent reliability, and their popularity among study participants, the computerized versions of the BASDAI, BASFI, BAS-G, and Quebec Scale seem preferable to the traditional paper format. PMID- 15801024 TI - Older adults' knowledge and beliefs about osteoporosis: results of semistructured interviews used for the development of educational materials. AB - OBJECTIVE: Although osteoporosis and associated fractures have been recognized as a significant public health problem, underdiagnosis and undertreatment are common. We investigated older adults' knowledge and beliefs regarding osteoporosis and its prevention, in order to develop effective osteoporosis health education messages and materials. These messages will be used as part of a trial that will test the efficacy of both public and doctor education to improve osteoporosis management. METHODS: We conducted semistructured one-on-one interviews with 15 older adult volunteers. A standard interview guide was developed and used for all interviews, which were audiotaped and transcribed. Key themes were extrapolated by 3 study staff using data abstraction forms. The data forms were then compared for consistency. RESULTS: We found that the term "osteoporosis" was well recognized, but many participants had only a fragmented understanding of its meaning. All participants identified osteoporosis as a serious condition, but many did not perceive themselves to be at personal risk for developing the condition. Many participants were confused about the difference between osteoporosis and osteoarthritis. Participants expressed reservations about taking prescription medications because of concerns over cost, side effects, and interactions with their current medications. CONCLUSION: Osteoporosis awareness is high, but the older adults interviewed had an incomplete understanding of the condition. This could hinder efforts to improve prevention and treatment of osteoporosis. PMID- 15801025 TI - Birthweight, vitamin D receptor gene polymorphism, and risk of lumbar spine osteoarthritis. AB - OBJECTIVE: To investigate risk factors for adult lumbar spine osteoarthritis (OA) including polymorphisms of the vitamin D receptor gene (VDR) and birthweight. METHODS: Plain radiographs of the lumbar spine were taken in 392 healthy subjects and graded for osteophytes and disc space narrowing (DSN); demographic data were collected. Details of birthweight and weight at 1 year were retrieved from historical records. VDR gene allelic variation was analyzed in 291 subjects. RESULTS: The mean age of the cohort was 65.8 years; mean weight was 68.9 kg in women and 80.1 kg and men. Osteophytes of grade >/= 2 were found in 63.5% of this cohort; DSN >/= 2 was present in 14.3% of subjects. Increasing osteophyte severity was significantly associated with age, adult weight, and manual social class; DSN was not. Presence and severity of osteophytes were associated with low birthweight and lower weight at 1 year in men, but not in women. No associations were found for DSN. The B allele of the VDR gene was associated with increasing severity of osteophyte. There was a significant interaction between birthweight and VDR gene in determining risk of osteophytosis in men (p for interaction = 0.04). The VDR-birthweight interaction pattern was similar but not statistically significant in women. CONCLUSION: Lumbar spine OA was a prevalent finding in this cohort. Both birthweight and polymorphisms in the VDR gene were associated with the presence of lumbar spine osteophytes and a significant interaction was observed between these 2 factors in men. PMID- 15801026 TI - Diagnosing hypovitaminosis D: serum measurements of calcium, phosphate, and alkaline phosphatase are unreliable, even in the presence of secondary hyperparathyroidism. AB - OBJECTIVE: To ascertain the ability of routine biochemical markers of bone turnover to predict vitamin D insufficiency. METHODS: Receiver operating characteristic (ROC) analysis was used to assess the value of serum alkaline phosphatase, calcium, and phosphate concentrations in the detection of hypovitaminosis D (< 20 nmol/l) in 467 patients between 1998 and 2000 (Cohort 1). The same analysis was repeated in a subsequent group of 719 patients between 2001 and 2003 (Cohort 2), in whom values of parathyroid hormone (PTH) were also available. Samples with elevated parathyroid levels from Cohort 2 were also analyzed to determine whether, in this subset, serum levels of calcium, phosphate, and alkaline phosphatase could reliably predict hypovitaminosis D. A subset of 50 patients from Cohort 1, with serum Vitamin D < 12 nmol/l, were reviewed by case note and telephone interview to determine demographic characteristics and the prevalence of risk factors for severe hypovitaminosis D. RESULTS: The areas under the ROC curves for alkaline phosphatase, calcium, and phosphate were all less than 0.7 (the criterion for a useful test) in both Cohorts 1 and 2. In the subset of Cohort 2 with elevated serum PTH levels (n = 337), the area under the ROC curve for calcium was 0.701 (95% confidence interval 0.643-0.758), and less than 0.7 for alkaline phosphatase and phosphate. In the 50 patients from Cohort 1 with severe hypovitaminosis D, risk factors were prevalent: 66% were vegetarian or vegan, clothing was partially or completely occlusive of sunlight (veiling) in 72%, and 60% of this cohort went outdoors less than 5 times per week. Symptoms were non-specific in the majority. CONCLUSION: Routine measurements of calcium, phosphate, and alkaline phosphatase are not reliable predictors of hypovitaminosis D, even when vitamin D insufficiency has been sufficient to produce a PTH response. Clinical suspicion based upon history and an awareness of risk factors should remain the gold standard for requesting serum vitamin D measurements. Inadequate sunlight exposure (through veiling and poor outdoor exposure) and poor dietary intake are highly prevalent features of hypovitaminosis D in severely affected patients. PMID- 15801027 TI - Inhibitors of hyaluronan export prevent proteoglycan loss from osteoarthritic cartilage. AB - OBJECTIVE: Osteoarthritis (OA) is characterized by cartilage erosion, proteolysis of aggrecan and collagen, and disturbed synthesis rates of aggrecan and hyaluronan by chondrocytes. The hypothesis is tested that hyaluronan overproduction contributes to aggrecan loss from osteoarthritic cartilage. METHODS: Human chondrocytes or bovine cartilage explants were incubated with interleukin 1beta (IL-1beta) to induce upregulation of hyaluronan and downregulation of aggrecan. OA was induced by injection of iodoacetate into the synovial cavity in rat knees. Hyaluronan export was inhibited by ATP-binding cassette transporter inhibitors such as the multidrug resistance (MDR) inhibitors valspodar or verapamil. The concentration of aggrecan was measured in cell culture media or visualized histochemically in cartilage tissue sections. RESULTS: Valspodar inhibited hyaluronan export from human chondrocytes in cell culture selectively without reducing aggrecan secretion. Valspodar and other MDR inhibitors prevented loss of aggrecan from osteoarthritic cartilage explants in culture. Verapamil prevented loss of aggrecan from cartilage in osteoarthritic rat knees. CONCLUSION: Hyaluronan is synthesized at plasma membranes and exported out of the cell. We recently identified an ATP-binding cassette transport system that is responsible for hyaluronan export. A number of ATP-binding cassette transport inhibitors are known and are in use clinically. These inhibitors were used here to inhibit hyaluronan export and to prevent aggrecan loss from arthritic cartilage. New drugs for treatment of arthritis are suggested by these studies. PMID- 15801028 TI - Cross-sectional association of 10 molecular markers of bone, cartilage, and synovium with disease activity and radiological joint damage in patients with hip osteoarthritis: the ECHODIAH cohort. AB - OBJECTIVE: To investigate the associations of molecular markers of joint tissue turnover with clinical and radiological variables in patients with hip osteoarthritis (OA). METHODS: Patients of the ECHODIAH trial cohort (60% female; mean age 63 yrs, disease duration 5 yrs) fulfilling the American College of Rheumatology criteria for hip OA were studied. Pain was assessed using a 100 mm visual analog scale, and the presence of night pain and morning stiffness was observed as the index of joint inflammation. Joint space width (JSW) and subchondral bone sclerosis were assessed on hip radiographs. Ten markers were measured, 8 in serum: N-propeptides of collagen type I (PINP) and type III (PIIINP), cartilage oligomeric matrix protein (COMP), YKL-40, hyaluronan (HA), matrix metalloproteases (MMP1 and MMP3), and ultrasensitive C-reactive protein (CRP); and 2 in urine: C-terminal crosslinking telopeptides of collagen type I (CTX-I) and type II (CTX-II). Analyses of 376 patients with measurements of all the markers included principal component analyses to identify independent clusters of markers; followed by stepwise multivariate regressions to determine associations between markers, clinical variables, and radiographic signs of joint damage. RESULTS: Markers could be segregated into independent clusters: CTX-II, PINP, and CTX-I for cartilage degradation and bone turnover; COMP, PIIINP, and HA as potential markers of synovitis; and CRP and YKL-40, which are likely to indicate systemic inflammation; plus MMP1 and MMP3. After adjustment for age, sex, and body mass index, pain was significantly associated with CTX-II (p = 0.0095) and CRP (p = 0.046) and joint inflammation with COMP (p = 0.013). Radiographic signs of joint damage were associated with CTX-II (p = 0.001 for JSW; p = 0.007 for bone sclerosis). CONCLUSION: This cross-sectional study of OA molecular markers in a large cohort may provide biological evidence of different pathophysiological processes involved in hip OA. Among the markers measured, CTX II showed the most consistent association with the symptoms and joint damage of OA. PMID- 15801029 TI - Leukotriene and prostaglandin synthesis pathways in osteoarthritic synovial membranes: regulating factors for interleukin 1beta synthesis. AB - OBJECTIVE: To study the mechanisms responsible for the cross-talk between lipoxygenase (LOX) and cyclooxygenase (COX) pathways in human osteoarthritic (OA) synovial explants, and to confirm the arachidonic acid (AA) shunting phenomenon and its influence on interleukin 1beta (IL-1beta) synthesis. METHODS: Synovial membrane explants were cultured in the absence or presence of different drugs that inhibit COX and/or LOX activities. Concentrations of prostaglandin E2 (PGE2), leukotriene B4 (LTB4), lipoxin A4 (LXA4), and IL-1beta were measured. RESULTS: When membrane explants were incubated with naproxen (COX inhibitor) under unstimulated conditions, the production of LTB4 was dose-dependently enhanced, reaching a 5-fold increase over the control. This shunt could be partially reversed by the addition of exogenous PGE2. Under lipopolysaccharide (LPS) stimulation, both licofelone (COX/LOX inhibitor) at therapeutic concentrations and NDGA (LOX inhibitor) inhibited LTB4 production, whereas naproxen did not amplify the LPS-induced LTB4 production. Conversely, using NDGA, it was found that a shunt of AA from the LOX to the COX pathway did not occur. Under LPS conditions, both naproxen and licofelone inhibited LXA4, inducing an increase in the LTB4/LXA4 ratio with naproxen treatment but not with licofelone. Under these conditions, naproxen treatment induced a higher level of IL-1beta production. CONCLUSION: We demonstrated in OA synovium that a shunt from AA to the LOX pathway occurred and that treatment with a nonselective COX inhibitor could increase the production of LTB4 and secondarily the synthesis of IL-1beta. Therefore treatment with licofelone, which can act on both COX and LOX pathways, may have some interesting properties in the treatment of OA. PMID- 15801030 TI - Interface tissue fibroblasts from loose total hip replacement prosthesis produce receptor activator of nuclear factor-kappaB ligand, osteoprotegerin, and cathepsin K. AB - OBJECTIVE: The highly osteolytic interface tissue between the bone and loosening total hip prosthesis is characterized by low pH, formation of foreign body giant cells, osteoclasts, and production of receptor activator of nuclear factor-kappaB (RANKL) and cathepsin K. We hypothesized that fibroblasts in the interface tissue may form a source for RANKL production. METHODS: Primary interface tissue fibroblasts, fibrous joint capsule fibroblasts, and trabecular bone osteoblasts were stimulated with tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), IL-6, IL-11, or 1alpha,25-(OH)2 vitamin D3. Cellular RANKL and released cathepsin K were detected by Western blotting. RANKL in cell lysates and osteoprotegerin (OPG) in cell culture medium were measured by ELISA. RANKL, OPG, and cathepsin K mRNA were measured with quantitative reverse transcriptase polymerase chain reaction. RESULTS: Interface tissue fibroblasts were found to produce RANKL. 1alpha,25-(OH)2 vitamin D3 stimulation increased RANKL mRNA expression. TNF-alpha was found to be the most potent OPG inducer in interface tissue fibroblasts. Cathepsin K mRNA production in fibroblasts was upregulated roughly 3-fold (p < 0.01) after 1alpha,25-(OH)2D3 stimulation, and both pro- and active cathepsin K protein was released to fibroblast culture media. CONCLUSION: Interface tissue fibroblasts are able to produce RANKL, OPG, and cathepsin K and may contribute indirectly and directly to pathologic periprosthetic collagenolysis and bone destruction. PMID- 15801031 TI - Employment across chronic inflammatory rheumatic diseases and comparison with the general population. AB - OBJECTIVE: To compare labor force participation across chronic inflammatory rheumatic diseases in order to assess the influence of the disease, disease duration, sex, education, and labor market conditions on employment. METHODS: Data from the German rheumatological database on outpatients of working age (20 59 yrs) between 1993 and 2001 were analyzed. The patients had rheumatoid arthritis (RA; n = 26,071), ankylosing spondylitis (AS; n = 5564), psoriatic arthritis (PsA; n = 6041), systemic lupus erythematosus (SLE; n = 4603), systemic sclerosis (SSc; n = 802), or Wegener's granulomatosis (WG; n = 385). Using population data, standardized employment ratios (SER) and part-time employment ratios of observed versus expected cases with 95% CI were calculated by means of indirect standardization for age and year of documentation. RESULTS: Across all diseases the overall employment rates were significantly lower than in the general population. Significant differences in SER were found between the diseases. The lowest SER of 0.76 to 0.81 (1.0 = population) were found in patients with RA, SLE, SSc, and WG. Higher SER were seen in AS (0.94) and PsA (0.92). In patients with a disease duration > 10 years the relative risk of being employed compared to RA, was 1.42 for AS, 1.26 for PsA, and 1.15, 1.03, 0.62 for PsA, SLE, SSc and WG, respectively. Comparing areas with low and high unemployment rates, a highly significant influence of labor market conditions on the SER was observed. The SER were significantly lower in patients with < 10 years of school education. CONCLUSION: Differences between employment rates in the population and the rates for the diseases under study are smaller than assumed by most clinical studies, especially in AS and PsA. However, these differences increase with longer disease duration. Specific measures to prevent patients from losing their job are needed, especially in areas with overall high unemployment. PMID- 15801032 TI - Bone mineral density in children and adolescents with systemic lupus erythematosus, juvenile dermatomyositis, and systemic vasculitis: relationship to disease duration, cumulative corticosteroid dose, calcium intake, and exercise. AB - OBJECTIVE: To describe the frequency of abnormal bone mineralization in a population of children with juvenile systemic lupus erythematosus (JSLE), juvenile dermatomyositis (JDM), and systemic vasculitis; and to investigate the relationship of bone mineral density (BMD) to cumulative corticosteroid dose, disease duration, Tanner stage, calcium intake, and exercise in these patients. METHODS: A retrospective chart review of children attending the pediatric rheumatology clinic at British Columbia's Children's Hospital was conducted to obtain demographic data (sex, ethnicity, disease duration, cumulative corticosteroid dose, and mean daily corticosteroid dose). All patients had at least one BMD measurement by dual energy x-ray absorptiometry (DEXA) at lumbar spine, hip, and total body. BMD was expressed as g/cm2 and Z scores; an abnormal Z score was defined as 0.85, and typical error was < 20% for the peak power and mean power on the WAnT. On the aerobic exercise test the R and ICC were > 0.95, and typical error was < 6% for the VO2peak and Wmax. CONCLUSION: We found acceptable reliability for the WAnT and very good reliability for the aerobic exercise test. This indicated that these exercise tests could be performed reliably in patients with JDM. PMID- 15801035 TI - A child with dermatomyositis and a suspicious lymphadenopathy. AB - Dermatomyositis (DM) in adults is frequently associated with cancer. In contrast, juvenile DM (JDM) is predominantly idiopathic and rarely reported with occult neoplasm. We describe a patient who presented with DM that was found to be a paraneoplastic manifestation of nasopharyngeal carcinoma. Although rare, paraneoplastic JDM must be suspected in the presence of unusual features such as elevated inflammatory markers or lymphadenopathy. Accurate clinical assessment including appropriate biopsies is needed before starting glucocorticoid therapy. PMID- 15801034 TI - Proliferative lupus nephritis and leukocytoclastic vasculitis during treatment with etanercept. AB - Tumor necrosis factor-alpha (TNF-alpha) is a proinflammatory cytokine. Agents that neutralize TNF-alpha are effective in the treatment of disorders such as rheumatoid arthritis, juvenile rheumatoid arthritis (JRA), spondyloarthropathies, and inflammatory bowel disease. TNF-alpha antagonist therapy has been associated with the development of antinuclear antibodies (ANA) and double-stranded DNA (dsDNA) antibodies, as well as the infrequent development of systemic lupus erythematosus (SLE)-like disease. We describe the first case of biopsy-confirmed proliferative lupus nephritis and leukocytoclastic vasculitis in a patient treated with etanercept for JRA. PMID- 15801036 TI - Intrafamilial variable phenotypic expression of a CIAS1 mutation: from Muckle Wells to chronic infantile neurological cutaneous and articular syndrome. AB - Among hereditary inflammatory disorders, Muckle-Wells syndrome, chronic infantile neurological cutaneous and articular syndrome (CINCA), and familial cold urticaria have recently been shown to be caused by dominantly inherited mutations in the CIAS1 gene. Reports suggest that these 3 diseases result from distinct missense mutations, with very few overlapping symptoms. We describe a French family presenting an intrafamilial overlapping clinical phenotype of CINCA and Muckle-Wells syndrome, caused by a mutation in CIAS1 gene. Clinical and genetic observations suggest that Muckle-Wells syndrome, CINCA, and familial cold urticaria are various phenotypic expressions of the same disease. PMID- 15801037 TI - Development of Crohn's disease in a patient taking etanercept. AB - In addition to its well known proinflammatory effects, tumor necrosis factor alpha (TNF-a) has complex effects on the growth, differentiation, and death of immune cells. TNF antagonists have had dramatic effects on the suppression of rheumatoid arthritis and other rheumatic inflammatory diseases. However, TNF inhibition of RA has led to an increased incidence of drug induced anti-dsDNA production, with cases of systemic lupus erythematosus as well as exacerbations of multiple sclerosis. While etanercept does not generally alter the course of Crohn's disease we describe a rare instance where this agent may have contributed to the development of clinically significant inflammatory bowel disease. PMID- 15801038 TI - Reversing myocardial microvascular disease in a patient with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is associated with increased cardiovascular mortality. This is regarded as being due to accelerated coronary atherosclerosis. We describe a 62-year-old man with seropositive erosive RA and extraarticular manifestations but no history of cardiovascular disease. Noninvasive assessment of myocardial blood flow by adenosine stressed thallium scanning showed reversible ischemia and diffusely poor myocardial perfusion. Repeat assessment after intensive immunosuppression for rheumatoid vasculitis revealed resolution of the ischemic changes and generally increased myocardial perfusion. Coronary angiography revealed no significant atheroma, suggesting that myocardial microvascular disease was responsible for the ischemia. This may be an important determinant of cardiovascular outcome in RA, and this case indicated that it can be reversed with immunosuppression. PMID- 15801039 TI - Isaacs' syndrome (autoimmune neuromyotonia) in a patient with systemic lupus erythematosus. AB - Patients with systemic lupus erythematosus (SLE) often produce autoantibodies against a large number of antigens. A case of SLE is presented in which muscle twitching and muscle cramps were associated with an autoantibody directed against the voltage-gated potassium channel of peripheral nerves (Isaacs' syndrome). PMID- 15801042 TI - Infliximab induced chilblain lupus in a patient with rheumatoid arthritis. PMID- 15801041 TI - Successful treatment of rheumatoid vasculitis-associated foot-drop with infliximab. PMID- 15801043 TI - Should vitamins be avoided early in life? PMID- 15801044 TI - On call. I've heard that men are making fewer sperm cells than they used to, and birth rates may suffer. What's the latest information? PMID- 15801045 TI - Abortion: time to clarify Australia's confusing laws. PMID- 15801046 TI - Throwing the baby out with the spa water? PMID- 15801047 TI - Substituting dietary linoleic acid with alpha-linolenic acid improves insulin sensitivity in sucrose fed rats. AB - This study describes the effect of substituting dietary linoleic acid (18:2 n-6) with alpha-linolenic acid (18:3 n-3) on sucrose-induced insulin resistance (IR). Wistar NIN male weanling rats were fed casein based diet containing 22 energy percent (en%) fat with approximately 6, 9 and 7 en% saturated fatty acids (SFA), monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) respectively for 3 months. IR was induced by replacing starch (ST) with sucrose (SU). Blends of groundnut, palmolein, and linseed oil in different proportions furnished the following levels of 18:3 n-3 (g/100 g diet) and 18:2 n-6/18:3 n-3 ratios respectively: ST-220 (0.014, 220), SU-220 (0.014, 220), SU-50 (0.06, 50), SU-10 (0.27, 10) and SU-2 (1.1, 2). The results showed IR in the sucrose fed group (SU-220) as evidenced by increase in fasting plasma insulin and area under the curve (AUC) of insulin in response to oral glucose load. In SU-220, the increase in adipocyte plasma membrane cholesterol/phospholipid ratio was associated with a decrease in fluidity, insulin stimulated glucose transport, antilipolytic effect of insulin and increase in basal and norepinephrine stimulated lipolysis in adipocytes. In SU-50, sucrose induced alterations in adipocyte lipolysis and antilipolysis were normalized. However, in SU-2, partial corrections in plasma insulin, AUC of insulin and adipocyte insulin stimulated glucose transport were observed. Further, plasma triglycerides and cholesterol decreased in SU-2. In diaphragm phospholipids, the observed dose dependent increase in long chain (LC) n-3 PUFA was associated with a decrease in LC-n-6 PUFA but insulin stimulated glucose transport increased only in SU-2. Thus, this study shows that the substitution of one-third of dietary 18:2 n-6 with 18:3 n-3 (SU-2) results in lowered blood lipid levels and increases peripheral insulin sensitivity, possibly due to the resulting high LCn-3 PUFA levels in target tissues of insulin action. These findings suggest a role for 18:3 n-3 in the prevention of insulin resistant states. The current recommendation to increase 18:3 n-3 intake for reducing cardiovascular risk may also be beneficial for preventing IR in humans. PMID- 15801048 TI - Plexiform neurofibromatosis involving the colon. PMID- 15801049 TI - Coronary interventions "to go": Is faster better? PMID- 15801050 TI - European workshop on MR and CT Imaging of the heart: Seville, Spain, October 2004. PMID- 15801051 TI - Basics of cardiac MDCT: techniques and contrast application. AB - The purpose of this article is to review the basic techniques of cardiac MDCT including the most recent 64-slice scanner generation. Besides the technical developments of different MDCT generations within recent years, the basic principles of ECG-related CT algorithms are discussed. Retrospective ECG-gated data acquisitions are considered as a prerequisite for reliable and reproducible calcium scoring as well as for volumetric assessment of the coronary artery tree. While data acquisition itself is rather straightforward, the optimization of image quality, the knowledge of adequate scan protocols and especially knowledge of possibly arising artefacts is mandatory for a stable high image quality. As data acquisition is becoming faster with the most recent advent of 64-slice MDCT, protocols have to be adapted to this new scanner generation. On the other hand, the improvement in spatial as well as in temporal resolution shows impact on the accuracy and stability of coronary CT angiography. However, the use of dose reducing technical improvements is strongly recommended. PMID- 15801052 TI - Basic techniques of cardiac MR. AB - Cardiovascular magnetic resonance (CMR) can be performed routinely due to the continued developments of MR systems, coil technology and sequence techniques, which have considerably shortened acquisition times. Techniques for the physiological monitoring of the patients are available for example, to synchronize the data acquisition with the individual heart cycle. The development of vectorcardiography-based (VCG) triggering improves the detection of the R waves and reduces the number of misinterpreted ECG events to a minimum. Depending on the clinical question, spin or gradient echo-based sequences with different k space filling methods are used in CMR. For spin preparation, inversion or saturation recovery techniques are used to suppress the signal of a specific tissue, or to maximize the T1 contrast between different soft tissues. Parallel acquisition methods (PAT) are available to speed up CMR imaging even further. PMID- 15801053 TI - Estimation of cardiac event risk by MDCT. AB - Coronary calcifications are specific markers for coronary atherosclerosis. The amount of coronary calcium is related to the likelihood of vulnerable plaques. Vulnerable plaques may rupture and may result in sudden coronary thrombus formation, occlusion, ischemia and ventricular fibrillation and finally cardiac death. Therefore, it is reasonable to believe that the risk of cardiac events can be assessed by the quantification of the extent of coronary calcium. However, until now, the predictive value of coronary calcium and the advantage over conventional risk factors has not yet been proven by any prospective cohort study. In practice uncertainty exists in the group of patients with an intermediate risk for cardiac events. In this particular cohort it is likely that the assessment of coronary atherosclerosis may help in the decision to initiate or discard a specific therapy. For this purpose it has been suggested to replace the Framingham age score by a score corrected by the amount of coronary calcium. Follow-up investigations may be helpful in the short term to determine the efficiency of different therapeutical options. To determine a significant progression of the amount of coronary calcium, the absolute mass should be determined in a period of 1 year. PMID- 15801054 TI - Assessment of cardiac function by CMR. AB - Cardiovascular magnetic resonance (CMR) has emerged as a valuable tool for the non-invasive morphological and functional assessment of the heart. By means of a variety of pulse sequences, accurate estimations of ventricular mass, volumes, ejection fraction and flow volume can be readily performed. When carefully prescribed, CMR sequences provide values on these parameters that can be considered as referential for both the right and left ventricle. This is due to the anatomical approach of the CMR method that, in contrast with other imaging techniques, is based on direct measurements rather than on indirect geometrical assumptions. PMID- 15801055 TI - Coronary CT angiography in symptomatic patients. AB - The currently best available spatial and temporal resolution for retrospectively ECG gated coronary multi-detector-row CT angiography is 0.4 mm and 165 ms, respectively. These acquisition parameters are already rather close to cardiac catheter. Studies so far compared non-invasive coronary CT and convention angiography for the detection of coronary artery stenoses. The most promising result reported by all authors was the high negative predictive value of the CTA. It now needs to be determined if CTA is a reliable tool to rule out coronary artery stenoses in a patient cohort with low likelihood of CAD, such as those with atypical chest pain or ambiguous stress test. CTA may furthermore establish as a rapid and widely available tool to detect vulnerable plaques or intracoronary thrombus in patients with acute coronary syndrome and unstable angina. In patients with chronic stable angina, tools that determine myocardial ischemia under stress such as SPECT and MRI are probably better suited to determine the relevance of coronary artery stenoses. In this particular cohort, by displaying the extent and morphology of coronary atherosclerosis, CTA may help to direct the therapy to either intervention or surgery. PMID- 15801056 TI - Role of CMR in assessment of myocardial perfusion. AB - Ischaemic heart disease is a major cause of morbidity and mortality. Assessment of myocardial perfusion is important in assessing the severity and extent of coronary artery lesions and in helping to guide revascularisation strategies where stenoses are documented. It provides information on the haemodynamic significance and a guide to prognosis. There has been much interest in the role of CMR assessment of perfusion and the opportunity for subendocardial resolution. We present an overview on its current status. PMID- 15801057 TI - Diagnosis of coronary artery disease with dobutamine-stress MRI. AB - Dobutamine-stress cardiovascular magnetic resonance (CMR) is a new diagnostic tool for the non-invasive detection of coronary artery disease. Technological advances in CMR have evolved this technique to an adequate alternative to the standard cardiac stress tests. Its high reproducibility and excellent image quality of the anatomical features of the left ventricle and left ventricular function at rest and during stress make it an ideal technique for the comprehensive evaluation of patients with suspected coronary artery disease. Besides its ability to detect myocardial ischemia, CMR has proved to be diagnostic for myocardial viability as well. A recent technical refinement in CMR using myocardial tagging has improved the diagnostic accuracy for myocardial ischemia even further. Dobutamine-stress CMR is used to identify wall motion abnormalities of the left ventricle in patients with proven or suspected coronary artery disease [1-4]. Dobutamine-stress CMR has emerged as a highly accurate and safe diagnostic modality [1-4]. Recently, the use of high-dose dobutamine CMR in combination with the myocardial tagging technique has been reported, with excellent diagnostic results. The use of this new technique and the clinical applications are discussed. PMID- 15801059 TI - Bypass graft imaging and coronary anomalies in MDCT. AB - Coronary artery bypass graft (CABG) is used to restore the blood flow in an ischemic area of myocardium using conduits bypassing a diseased coronary artery. Until now, conventional angiography has been the recognized technique to study patients with bypass grafts. Nowadays, non-invasive methods such as multi detector CT (MDCT) emerge as reliable imaging methods in the study of CABG. Thus, radiologists play an important role in this field, not only to define if the bypass graft is occluded or stenosed but also to report further information such as CABG technique performed, type of conduit used or pre-operative findings. This paper analyzes the most practical information that the radiologist must know in a study of CABG. Another theme which will be briefly described is the use of MDCT in coronary anomalies studies, with particular emphasis on the course of the abnormal vessel and its relation to great vessels. PMID- 15801058 TI - Delayed contrast-enhanced MRI: use in myocardial viability assessment and other cardiac pathology. AB - As in other organs, tissue characterization is important for many cardiac diseases. For example, in ischemic heart disease, differentiation between reversibly and irreversibly damaged myocardium in patients with a prior myocardial infarction is crucial in determining disease severity, functional recovery and patient outcome. With the recent advent of the single inversion recovery contrast-enhanced magnetic resonance imaging (MRI) sequence (delayed contrast-enhanced MRI), contrast between normal and abnormal tissues could be significantly enhanced compared with the conventional cardiac MRI sequences, enabling even subtle abnormalities to be visualized. Together with other advances in cardiac MRI (e.g. functional imaging, coronary artery imaging), MRI has become one of the preferred non-invasive modalities to study cardiac diseases. In this paper an overview of the versatility of delayed contrast-enhanced MRI for investigating cardiac diseases is given. PMID- 15801060 TI - Another dose of dose. PMID- 15801061 TI - Autosomal dominant polycystic kidney disease presenting as subarachnoid hemorrhage. AB - Intracranial aneurysms occur in patients with autosomal dominant polycystic kidney disease (ADPKD) approximately five times more often than in the general population, and in the same patient group, subarachnoid hemorrhage from rupture of aneurysms occurs about a decade earlier than in the general population. We present a case of unsuspected ADPKD presenting as spontaneous subarachnoid hemorrhage from a ruptured intracranial aneurysm. PMID- 15801062 TI - [Life-threatening complications after plastic tracheostomy closure. Which length of hospital stay according to clinical and forensic viewpoints is necessary?]. AB - BACKGROUND: Plastic tracheostomy closure represents a safe surgical method with rare but potentially life-threatening complications such as dyspnea. Because of the general tendency to reduce hospital stay, the medically necessary time of hospitalization should be evaluated. METHODS: Ninety-eight patients operated in succession were examined retrospectively for extent, time, and therapeutic procedures in cases of life-threatening dyspnea. Moreover, disorders of wound healing which had to be treated surgically were analyzed. RESULTS: Of 98 patients 17% (17/98) developed dyspneas, 10% (10/98) up to the 3rd postoperative day, 2% (2/98) on the 6th postoperative day, and 5% (5/98) between the 17th and 92nd postoperative days; 13% (13/98) had to be treated because of extensive disorders of wound healing, 85% (11/13) up to the 3rd day. CONCLUSION: From the clinical and forensic viewpoint, a hospital stay for at least 3 postoperative days is necessary to record the majority of life-threatening complications after plastic tracheostomy closure. PMID- 15801063 TI - [The anterosigmoidal approach. A function-preserving surgical method for petroclival tumors]. AB - BACKGROUND AND OBJECTIVE: The antero- or persigmoid approach preserves all functional structures of the petrous bone and, therefore, is an alternative to the classic laterobasal approaches for exploring the petroclival region. As high morbidity is assumed and it is a time consuming procedure, this approach is not well known. METHODS AND PATIENTS: Initially, a cadaver study using computer assisted surgery was used to determine the surgical workflow before we optimized our surgical procedures. RESULTS: In a series of 7 patients with benign tumors growing from the petrous apex to the petroclival region, we were able to resect all tumors completely via the anterosigmoidal surgical corridor using computer assisted surgery. CONCLUSIONS: The navigation-guided anterosigmoidal approach is an excellent method, allowing a good overview of the petroclival region. In patients suffering from complex petroclival tumors, it offers a real chance to achieve complete microsurgical tumor resection without functional defects by an interdisciplinary neuro-oto-surgical session. PMID- 15801064 TI - [Chronic hyperplastic laryngitis following treatment of hypertension with angiotensin converting enzyme-inhibitor]. AB - We report the case of a 42 year old patient who developed chronic hyperplastic laryngitis during treatment with the angiotensin converting enzyme-inhibitor Cibacen 10. A severe cough and vocal restrictions with hoarseness were only incompletely cured after changing this anti-hypertensive medication to a adrenergic blocker, combined with a vocal rest and anti-inflammatory inhalation. Therefore we performed a laryngoscopy under general anesthesia and excised the swelling of the vocal cords. Additionally, voice therapy was prescribed and complete restitution achieved. Although hoarseness is documented as a potential side effect of angiotensin converting enzyme-inhibitors, morphological alterations in the vocal cords have not been linked to this type of drug. In our case, prolonged medication with Cibacen 10 led to chronic hyperplastic laryngitis. Initial coughing might have induced the trauma of the epithelium of the vocal cords. Due to the morphological alterations to the vocal cords the patient developed additional functional dysphonia. PMID- 15801065 TI - [Cystic myoepithelioma. A rare differential diagnosis of a cystic lesion of the parotid gland]. AB - By sonography, we found a sharply demarcated tumor with cystic areas in the parotid gland of a 41 year old male, indicating Warthin's tumor. Subtotal parotidectomy was performed. Microscopy showed an encapsulated tumor with myoepithelial cells and, in particular, central pseudocysts. Immunohistochemically, the tumor cells expressed cytokeratin 5/6 and S-100 protein as well as smooth muscle-actin. These features led to the diagnosis of a cystic myoepithelioma. Histopathologically, several different lesions of the salivary glands should be considered in the differential diagnosis of myoepithelioma, especially of this hitherto unique case in the parotid gland. The differential diagnoses are reviewed and discussed. Treatment is by surgical resection. Because of the tendency of myoepitheliomas to recur and to malignant transformation, tumor-free margins are recommended. PMID- 15801066 TI - [The cochlear implant. Molecular arguments favouring early implantation]. AB - BACKGROUND: An important factor in the clinical outcome of cochlear implantation is the age of the patient. Compared to older patients, children with congenital deafness have a better outcome when the implantation is made before the age of 2 years. The cause may lie in the molecular biology of the brain, which changes during postnatal maturation. METHODS: Protein probes were obtained from tissue of the rat inferior colliculus at different ages. The probes were analyzed using 2 dimensional SDS electrophoresis. RESULTS: The expression of GAP-43, a protein expressed by neurons during axonal outgrowth and synaptogenesis, and the total number of the protein species showed a significant reduction during ontogenesis. This shows that while neurons gradually assume their specific function, they downregulate GAP-43 and the molecular complexity decreases. CONCLUSIONS: Due to a lack of neuronal pluripotency at later developmental stages, the flexibility to adapt to the afferent activation provided by a cochlear implant is increasingly limited. PMID- 15801067 TI - Diagnostic performance of anti-beta2 glycoprotein I and anticardiolipin assays for clinical manifestations of the antiphospholipid syndrome. AB - The objective of the present study was to analyse the performance of the tests for detection of anti-beta2 glycoprotein I (beta2 GP I) and anticardiolipin (aCL) antibodies for identification of clinical manifestations of the antiphospholipid syndrome (APS). Patients with systemic lupus erythematosus (SLE) as well as carriers of infectious diseases such as Kala-azar, syphilis and leptospirosis were studied. Particular interest was given to the presence of clinical complications related to APS. Anticardiolipin and anti-beta2 GP I antibodies were searched using an enzyme-linked immunosorbent assay (ELISA) assay. Clinical manifestations of APS were observed in 34 of the 152 patients (22.3%) with SLE and no patient with infectious disease had such manifestations. Antibodies to cardiolipin in moderate or high levels and anti-beta2 GP I were detected in 55 of 152 (36.1%) and 36 of 152 (23.6%) patients with SLE, respectively, and in 2 of 30 (6.6%) and 16 of 30 (53.3%) patients with Kala-azar, in 9 of 39 (23%) and 6 of 34 (17.6%) patients with leptospirosis, and 14 of 74 (18.9%) and 8 of 70 (11.4%) cases of syphilis, respectively. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and likelihood ratio (LR) of the anti-beta2 GP I test for the identification of the clinical manifestation of APS were, respectively, 29% [95% confidence interval (CI) = 24%-34%], 78% (95% CI = 73-83%), 15% (95% CI = 11-19%), 89% (95% CI = 85-93%) and 1.38. Regarding the aCL assay, the figure was 29% (95% CI = 24-34%), 76% (95% CI = 71-81%), 14% (95% CI = 10-18%), 89% (95% CI = 86-92%) and 1.26. As the validity and performance of the anti-beta2 GP I assay were similar to the aCL in demonstrating the presence of clinical phenomena associated with APS and due to the difficulties in performing as well as the lack of standardisation of the anti beta2 GP I test, we suggest that the test for aCL should continue to be the first one performed when the presence of APS is suspected. PMID- 15801068 TI - Osteocalcin synthesis by human osteoblasts from normal and osteoarthritic bone after vitamin D3 stimulation. AB - Alterations in osteoblast metabolism are involved in the pathogenesis of typical subchondral bone changes in osteoarthritis (OA). Osteocalcin is a specific bone protein, synthesised by the osteoblasts, which can be considered a marker of metabolic activity of these cells. In this study we correlated osteocalcin production from human osteoblasts isolated from healthy and osteoarthritic subjects to the degree of cartilage damage, before and after stimulation with 1,25(OH)2-vitamin D3, the active metabolite of vitamin D3. We isolated human osteoblasts from cancellous bone of healthy subjects and from subchondral bone of osteoarthritic subjects and considered the osteoblasts corresponding to different degrees of cartilage damage as different cell populations. We determined the osteocalcin production in normal and osteoarthritic osteoblasts from maximal and minimal cartilage damage areas both under basal conditions and after vitamin D3 stimulation. Compared to normal osteoblasts, under basal conditions osteocalcin production is significantly greater in osteoarthritic osteoblasts, corresponding both to maximal and minimal damage joint areas. No differences were observed between osteoblasts from maximal and minimal damage areas. The response of osteoblasts to vitamin D3 stimulation appeared to be proportional to the degree of joint damage, as the vitamin D3-induced increase in osteocalcin is proportionally greater in maximally damaged osteoblasts compared to minimally damaged ones. Thus, after vitamin D3 stimulation, a significant increase in osteocalcin production by maximally damaged osteoblasts compared to the minimally damaged ones was observed. This study confirms abnormal osteoarthritic osteoblast behaviour and indicates that osteoblasts from different areas of the same affected joint may be metabolically different, supporting the hypothesis that subchondral osteoblasts may play an essential role in the pathogenesis of OA. PMID- 15801069 TI - Clinical features of ten cases of osteopoikilosis. AB - A retrospective study was carried out of the ten cases of osteopoikilosis seen at this Orthopedic Unit over a 15-year period in order to determine the reasons why patients seek consultation, preliminary diagnosis, and associated lesions. Eight patients consulted for problems not related to the locomotor apparatus, and diagnosis was by chance; the other two presented joint pain. The preliminary diagnosis was osteoblastic metastasis in five patients and osteopoikilosis in the other five. None of the patients displayed skin or visceral involvement, but three presented bone alterations. Definitive diagnosis was made by measurement of biochemical markers of bone remodeling, radiography of both hands, and bone scan. Bone biopsy was performed in one case. Although rare, the radiographic symptoms of osteopoikilosis are sufficiently specific to avoid false diagnoses, which may give rise to rigorous and expensive studies for other important disorders. PMID- 15801070 TI - Utility of age, gender, ANA titer and pattern as predictors of anti-ENA and dsDNA antibodies. AB - Monovariate and multivariate analyses including logistic regression were performed to determine associations among predicting variables [age, gender, immunofluorescence pattern, and anti-nuclear antibody (ANA) titer] and anti extractable nuclear antigen (ENA) and -dsDNA antibodies (Abs) in 1089 patients with positive fluorescent ANA (FANA) test results. Samples with high titer ANAs had an increased frequency of anti-ENA and -dsDNA Abs. The receiver operating (ROC) curves of the ANA titer for anti-ENA Abs had a larger under the curve area compared to the ROC curve for anti-dsDNA Abs, indicating that ANA titer is better for predicting anti-ENA Abs than anti-dsDNA Abs. There was no relation noticed between immunofluorescence patterns and anti-ENA and -dsDNA Abs except an increased frequency of anti-dsDNA Abs found in samples with a homogeneous pattern. Probability calculations on the basis of the ANA pattern and the titer showed that samples with low titer ANAs (1:160 or less) had low probabilities for anti-ENA Abs (0.002-0.009) regardless of immunofluorescence patterns. However, samples with a homogeneous pattern at any titers including low titers had high probabilities for anti-dsDNA Abs. A decreased frequency of anti-dsDNA Abs as measured by Crithidia assay was noticed in samples from patients aged 50 or older. In contrast, no association was noticed between age and anti-ENA Abs. There was no female preponderance found in the presence of anti-ENA and -dsDNA Abs. In conclusion, our study shows that the ANA titer but not the immunofluorescence pattern is useful in predicting anti-ENA Abs. In contrast, both the ANA titer and the immunofluorescence pattern help in predicting anti dsDNA Abs. Samples with low titer ANAs (1:160 or less) may not need a further test for anti-ENA Abs unless an ANA-associated disease is highly suspected. However, a test for anti-dsDNA Abs should be considered in samples with a homogeneous pattern at any titer including low titers. PMID- 15801072 TI - Intravenous immunoglobulin in treatment of cardiac tamponade in a patient with systemic lupus erythematosus. AB - We describe a 23-year-old female patient with a history of systemic lupus erythematosus and pulmonary hypertension who developed a large pericardial effusion with cardiac tamponade. Invasive interventions such as pericardial window or pericardiectomy were ruled out because of the posterior localization of the effusion and high risk of general anesthesia in a patient with severe pulmonary hypertension. The patient received high-dose steroids intravenously with no response. A 5-day course of intravenous immunoglobulin resulted in gradual decrease of the pericardial effusion and resolution of cardiac tamponade within 2 weeks. PMID- 15801071 TI - Association between systemic sclerosis and breast cancer: eight new cases and review of the literature. AB - Several studies have demonstrated an increased frequency of cancer in patients with systemic sclerosis (SSc), specially lung and breast cancers. The pathogenesis of the association between SSc and cancer is not fully established. The aim of this study was to describe new cases of the association between SSc and breast cancer and to perform a review of the literature. We retrospectively studied the medical files of eight patients followed in our institution for SSc and breast cancer. We analyzed them with data available in the literature for a total of 46 patients. Cutaneous extension of SSc was clearly mentioned in 17 cases: the SSc was limited in 10 cases and diffuse in 7 cases The median age at the diagnosis of cancer was 54 years (range: 40-71). The median duration between SSc onset and breast cancer diagnosis was 11.5 months (range: 0-288). The duration between SSc onset and breast cancer diagnosis was < or = 12 months in 27 of 44 patients (61.4%), and in 11 (25%) of them the diagnosis of both diseases was made simultaneously. It was clearly mentioned for 35 patients whether the diagnosis of breast cancer was made before or after the onset of SSc. The diagnosis of breast cancer was made before SSc onset in 17 of 35 patients (48.6%) and after SSc onset in 18 of 35 patients (51.4%). For 33 patients, the follow-up was available: 18 (54.5%) died, 11 (33.3%) of them within the 1st year after the diagnosis of the cancer. For none of the patients did the anticancer treatment improve the SSc. The close temporal relationship between SSc onset and breast cancer diagnosis is highly suggestive of a pathophysiological link. SSc is probably not a paraneoplastic disease since the anticancer treatment has no influence on the evolution of SSc. However, it can be suggested that SSc could be a disease facilitating breast cancer and/or metastases development. PMID- 15801073 TI - Pregnancy in Wegener's granulomatosis: successful treatment with intravenous immunoglobulin. AB - We present the case of a women diagnosed with Wegener's disease at the age of 26 years, refractory to corticosteroid and cyclophosphamide therapy. Treatment with intravenous immunoglobulin (IVIg) was started, leading to partial clinical remission of disease. During IVIg treatment she became pregnant. IVIg therapy was continued, the disease went into remission, and after 40 weeks the patient delivered a healthy boy. After 6 months from the delivery, the patient became pregnant again. Now she is at the 22nd week of pregnancy and she is doing very well. This case supports the beneficial effect of IVIg in Wegener's granulomatosis and illustrates its safety and efficacy during pregnancy. PMID- 15801074 TI - A case of Takayasu arteritis complicated with glomerulonephropathy mimicking membranoproliferative glomerulonephritis. AB - We report on a 65-year-old Japanese woman with Takayasu arteritis who developed severe proteinuria, hypertension, and renal dysfunction. Renal angiography demonstrated moderate irregular narrowing of both renal arteries. Renal biopsy showed glomerulonephropathy mimicking membranoproliferative glomerulonephritis (MPGN) with glomerular capillary wall thickening ("double contour") accompanied by mesangial cell proliferation and moderate increase of mesangial matrix without deposits of C3. Electron microscopy showed no subendothelial deposit and no circumferential mesangial interposition (CMI), and these findings are different from MPGN. In this report we present a case of Takayasu arteritis associated with glomerulonephropathy mimicking MPGN. PMID- 15801075 TI - Intravenous cyclophosphamide therapy in a case with refractory thrombotic microangiopathic hemolytic anemia and SLE. AB - The case of a 27-year-old woman who simultaneously presented with SLE and severe refractory thrombotic microangiopathic hemolytic anemia (TMHA) is reported. She had extremely high levels of platelet-associated IgG (PAIgG), and her TMHA was refractory to plasma exchange and corticosteroid therapy. However, the TMHA was effectively controlled by i.v. cyclophosphamide therapy. ITP and TTP are generally considered distinct diseases; however, TMHA may occur secondary to platelet aggregation via autoimmune mechanisms in certain cases. Immunosuppressive therapy at an early stage of the disease may be beneficial in refractory cases of TMHA with autoimmune features. PMID- 15801076 TI - A handicap in treatment and follow-up of ankylosing spondylitis: sports hematuria. AB - A 19-year-old male patient with a 7-year history of ankylosing spondylitis (AS) was admitted to our clinic. After completion of the laboratory and radiologic investigations, we prescribed an exercise program besides his medical therapy. After these exercises, he complained of painless dark urine sometimes with passage of clot that disappeared the following day. To investigate this hematuria related with exercise, further laboratory and radiologic studies were carried out. After exclusion of the other causes of hematuria, we concluded that the diagnosis was sports hematuria. PMID- 15801077 TI - Severe carpal tunnel syndrome in a patient with juvenile idiopathic arthritis due to proximal migration of hypertrophic lumbrical muscles. AB - We report a new case of pediatric carpal tunnel syndrome in a patient with juvenile rheumatoid arthritis. Symptoms were mainly motor weakness and severe atrophy of the thenar. PMID- 15801079 TI - Dermatomyositis and duodenal carcinoid. PMID- 15801078 TI - Pulmonary coinfection by Pneumocystis carinii and Aspegillus fumigatus in a seronegative arthritis patient treated with low-dose methotrexate. PMID- 15801080 TI - A case of recurrent pancreatitis due to hyperlipidemia misdiagnosed as familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is prevalent among Arabic, Turkish, Armenian, and Jewish people and it must always be considered in the differential diagnosis of patients from these ethnic groups presenting with recurrent abdominal pain with fever. In cases of fever and recurrent abdominal pain, acute pancreatitis is an important clinical condition, which should be considered in the differential diagnosis. Serum amylase concentration in acute pancreatitis is usually more than three times the upper limit of normal. However, in recurrent pancreatitis secondary to hypertriglyceridemia, serum amylase levels, for reasons that are not well understood, may be normal or mildly elevated. Recurrent pancreatitis secondary to hypertriglyceridemia may thus pose a problem in the differential diagnosis and may lead to an erroneous diagnosis of FMF. Measurement of serum triglyceride along with amylase levels should be required for a suspected diagnosis. Computerized examination of the abdomen may need to be undertaken to exclude acute pancreatitis in the presence of hypertriglyceridemia since serum amylase levels may be normal or slightly elevated. PMID- 15801081 TI - HIV infection mimicking SLE. PMID- 15801082 TI - Re: "Coexisting seronegative rheumatoid arthritis and ankylosing spondylitis". PMID- 15801083 TI - Endotoxin as a factor for joint pain and rheumatoid arthritis. PMID- 15801084 TI - More research on women's unique heart risks. Now that studies of heart disease include women, we're learning about "heart-felt" sex differences. PMID- 15801085 TI - Treating rheumatoid arthritis. Just opening a jar can be a harsh reminder of the limitations rheumatoid arthritis places on daily life. Early and aggressive treatment can often help. PMID- 15801086 TI - Light therapy for winter depression. If you suffer from seasonal affective disorder, you don't need to wait for longer days to get some relief. PMID- 15801087 TI - Bad actor in cardiovascular disease may play role in fractures. PMID- 15801088 TI - By the way, doctor. Which calcium supplement is better for older women, calcium citrate or calcium carbonate? PMID- 15801089 TI - By the way, doctor. A preventive doctor told me to take indolplex (bioavailable diindolylmethane or DIM) to aid estrogen metabolism. Is there any research available about this supplement? PMID- 15801090 TI - Five for 2005: five reasons to forgive. PMID- 15801091 TI - Gynecologist or internist? PMID- 15801092 TI - Anesthesia and how to prepare for it. Understanding your options and knowing what to expect will increase your chances of having a smooth surgical experience and recovery. PMID- 15801093 TI - Ultra-low dose estrogen patch improves bone, appears safe for the uterus. PMID- 15801094 TI - Ductal lavage not a good screening test for breast cancer. PMID- 15801095 TI - By the way, doctor. I get cold sores every few months. I'm worried I'll have them when I'm away on vacation next month. What can I do to prevent them? PMID- 15801096 TI - Kitchen scalds: a poorly recognized hazard to young children. PMID- 15801097 TI - New autism campaign launched this month. PMID- 15801098 TI - "Hypnosis" can help children with painful medical procedures. PMID- 15801099 TI - Child safety. 15-passenger vans can pose special risks. PMID- 15801100 TI - Product recalls. Children's costume bracelets...and children's necklaces. PMID- 15801101 TI - The elephant from the other side of silence (or HIV news from four compass points). XV International AIDS Conference, Bangkok, 11-16 July 2004. PMID- 15801103 TI - High time to vastly scale up action on human rights. PMID- 15801102 TI - Focus on hepatitis. Final APRICOT, ACTG A5071 results. PMID- 15801104 TI - Still underground: searching for progress in realizing the human rights of women in prostitution. PMID- 15801105 TI - Steps forward, backward, and sideways: Canada's bill on exporting generic pharmaceuticals. AB - In May 2004 Canada's Parliament passed Bill C-9, amending the Patent Act to provide for the compulsory licensing of patented pharmaceutical products. The bill allows generic manufacturers to make cheaper, generic versions of patented products and export them to countries that do not have sufficient capacity to produce their own. Canada thus became the first country to pass legislation implementing a World Trade Organization (WTO) Decision that relaxed WTO rules on pharmaceutical patents. Civil society organizations campaigned to enhance the bill before it passed, and succeeded in obtaining significant improvements. However, the final bill is marred by flaws that mean it falls short of being a "model" that should simply be replicated elsewhere. This article is the second in a series tracing the evolution of Canada's legislation. It reviews the developments leading up to the adoption of Bill C-9 in its final form, and analyzes its positive and negative aspects. Hopefully, other advocates can learn from this experience and other countries can avoid replicating the negative aspects as they implement the WTO Decision. PMID- 15801106 TI - Ontario: police disclose HIV status of accused under Police Services Act. AB - In Ontario, people suspected of or charged with aggravated assault for exposing another person to HIV through unprotected sexual intercourse risk not only a criminal conviction but also having their names, dates of birth, descriptions, photographs, addresses, sexual history, employment information, and HIV status made public. This disclosure without the consent of the person suspected or charged has usually taken the form of a "public safety advisory." To date, at least five such advisories have been released by police services across Ontario. PMID- 15801107 TI - BC: campaign launched to protect personal health information. AB - A diverse group of rights, health, union, and HIV/AIDS organizations has launched a province-wide campaign to demand that the British Columbia government cease contracting out the administration of its medical plans to a private US company. The Right to Privacy Campaign (RPC) believes that the government's contract with Maximus Inc. places British Columbians' confidential health and related information within easy reach of US government agencies as a result of provisions in the USA PATRIOT Act and other US legislation. PMID- 15801108 TI - Nova Scotia: "blood samples" legislation passed. AB - On 18 October 2004 the Nova Scotia legislature passed the Mandatory Testing and Disclosure Act, thus becoming the third Canadian province or territory to pass similar legislation. PMID- 15801109 TI - British Columbia: studies show positive public impacts of harm-reduction measures for drug users. AB - Two recent studies from the BC Centre for Excellence in HIV/AIDS show that HIV prevention measures for IV drug users increase public order, and can potentially save substantial public resources. PMID- 15801110 TI - Nurse, legal society receive human rights award. AB - The 2004 Canadian Awards for Action on HIV/AIDS and Human Rights have been presented to Megan Oleson, a Vancouver nurse who set up a temporary, unauthorized safe injection site (SIS) for intravenous drug users in the Downtown Eastside; and to the Pivot Legal Society (PLS), also of Vancouver. The PLS worked with Oleson to set up the temporary site while Canada's first legal SIS was still under construction. PMID- 15801111 TI - Proposed amendments to medical marijuana regulations released for comment. AB - On 23 October 2004, Health Canada released proposed amendments to the Marihuana Medical Access Regulations (MMAR) for public comment. The amendments are intended to respond to consumer concerns about the onerous bureaucratic requirements for obtaining legal permission to possess and cultivate marijuana for medical purposes, and to address the need for a safe, reliable, legal source of medical marijuana. Under the proposed amendments, police forces throughout Canada will be able to access limited information about people authorized to possess marijuana, people licensed to produce marijuana, and legal grow operations. PMID- 15801113 TI - Vancouver: gay posters banned by billboard company. PMID- 15801112 TI - MP calls for review of solicitation laws. PMID- 15801114 TI - Groups distribute harm-reduction kits to crack users. PMID- 15801115 TI - British Columbia study finds high levels of poverty and disability among HIV positive. PMID- 15801116 TI - HIV/AIDS, law, and discrimination in Guyana. AB - In June 2004 the Guyanese National AIDS Committee completed a comprehensive national assessment on HIV/AIDS, law, ethics, and human rights - the first to be completed under the Pan-Caribbean Partnership against HIV/AIDS (PANCAP) Project on Law, Ethics and Human Rights. In this article the author of the report, Arif Bulkan, reviews the application of Guyanese law to HIV/AIDS-related discrimination, and notes the considerable gap between law and practice. PMID- 15801117 TI - AIDS out of focus? PMID- 15801118 TI - Is PEPFAR tackling HIV drug supply in wrong way? PMID- 15801119 TI - State of AIDS care 2004: one world, multiple standards. July 15, 2004 - Bangkok. PMID- 15801120 TI - Hepatitis coinfection and LPV/RTV. PMID- 15801122 TI - Abstracts from the 4th European Congress on Nutrition and Health in the Elderly. November 4-5, 2004. Toulouse, France. PMID- 15801121 TI - Proceedings of the Fourth International Cytoprotection Investigators' Congress. May 2004. Philadelphia, Pennsylvania, USA. PMID- 15801123 TI - Proceedings of an international conference "Targeting RNA: artificial ribonucleases, conformational traps and RNA interference" and a workshop "Biology and biochemistry of extracellular nucleic acids." June 18-21, 2003. Novosibirsk, Russia. PMID- 15801124 TI - Each genomic RNA in HIV-1 heterozygous virus generate new virions. AB - Retrovirus are unique because they present two complete copies of the genomic RNA in each virion. It is believed that only one proviral DNA is formed from the two genomic RNAs. To check this hypothesis, we constructed two deleterious HIV-1 variants in gag gene which upon transfection in Cos-1 cells were able, by complementation, to form heterozygous viruses, used to infect MT4 cells in a plaque assay. Analysis of the proviral DNA of the eight plaques obtained indicated that five were recombinants between the two deleterious mutants. Three other plaques showed three bands corresponding to the reverse transcription of both strands of one heterozygous virion and to the recombination of the two genomes. These results demonstrate that the two genomic RNAs in HIV-1 heterozygous virions could be used in the generation of new viruses. This mechanism permits the recovery of deleterious mutants and enhances the evolutive potential of HIV-1. PMID- 15801125 TI - Proceedings of the Twenty-Third International Symposium on the Separation of Proteins, Peptides, and Polynucleotides. November 9-12, 2003. Delray Beach, Florida, USA. PMID- 15801126 TI - Cellular evidence of allelopathic interference of benzoic acid to mustard (Brassica juncea L.) seedling growth. AB - Cellular changes in the roots of mustard (Brassica juncea L.) grown in soil treated with 1.09, 1.46 and 1.83 mg benzoic acid per g soil, a known allelochemical, were analyzed after 7 days. The recoverable concentration of 1.09, 1.46 and 1.8 mg benzoic acid per g soil (measured by high performance liquid chromatography) was 68, 150 and 250 microg benzoic acid per g soil, respectively. The benzoic acid treatments suppressed root growth by 30.5%, 58.8% and 81.1% with increasing concentrations. Transmission electron microscopy studies of roots showed irregular shaped cells arranged in disorganized manner and disruption of cell organelles at cellular level. Root cells showed dissolution of middle lamella (at 68 and 150 microg benzoic acid per g soil) but intact middle lamella with increased wall deposits was observed with 250 microg benzoic acid per g soil. Damage to the mustard root at cellular level was evidenced by changes in cell morphology and internal organization. PMID- 15801127 TI - Proceedings of the First International Society of Chemotherapy International Conference on Cancer Therapeutics-Molecular Targets, Pharmacology and Clinical Applications. February, 19-21, 2004. Florence, Italy. PMID- 15801128 TI - Abstracts from the XVIIth International Symposium on Technological Innovations in Laboratory Hematology. May 13-15, 2004. Barcelona, Spain. PMID- 15801129 TI - Festschrift in honor of the 60th birthday of Bill Hardcastle. PMID- 15801130 TI - Proceedings of the 9th Central European Congress of Coloproctology and the 4th Symposium of Coloproctology of Yugoslavia. Belgrade 7-9 October 2004. PMID- 15801131 TI - JournalScan. PMID- 15801132 TI - Nonpayment of benefits when the Social Security Administration receives notice that an insured person is departed or removed from the United States. Final rule. AB - To implement part of the Social Security Protection Act of 2004 (SSPA), we are revising our regulations that prohibit payment of monthly benefits and the lump sum death payment under title II of the Social Security Act (the Act) when SSA receives notice that an insured person is deported or removed from the United States under certain provisions of the Immigration and Nationality Act (INA). PMID- 15801133 TI - Regarding the case report published recently. PMID- 15801134 TI - Using patent foramen ovale (PFO) for percutaneous balloon mitral valvuloplasty. PMID- 15801135 TI - Stenting of anomalous right coronary artery. PMID- 15801136 TI - Festschrift in memory of Dr. Herbert Remmer. PMID- 15801137 TI - ["Euthanasia" debates in Germany (1895-1945)]. PMID- 15801139 TI - South Asia slashes polio cases by nearly half. PMID- 15801138 TI - Complications of cervical spine manipulation therapy: 5-year retrospective study in a single-group practice. PMID- 15801140 TI - Global tobacco treaty enters into force with 57 countries already committed. PMID- 15801141 TI - CDC launches "Learn the Signs, Act Early." campaign. PMID- 15801142 TI - CDC breaks ground on new lifestyle center, dedicates new streetscape. PMID- 15801143 TI - HIV vaccine global partners strengthen collaboration to speed up progress. PMID- 15801144 TI - Coast-to-coast polio drive to counter epidemic in Africa. PMID- 15801145 TI - Sagittal alignment and the Bryan cervical artificial disk. PMID- 15801146 TI - Sagittal alignment and the Bryan cervical artificial disk. PMID- 15801147 TI - PPAR activators and gastroenterology. PMID- 15801148 TI - [A virtual reality based navigation system for endovascular therapy training]. PMID- 15801149 TI - [Troponin Ic and renal insufficiency]. PMID- 15801150 TI - [Cryoglobulinemia: updating]. PMID- 15801151 TI - Guidelines on confidentiality for population-based cancer registration. PMID- 15801152 TI - International rules for multiple primary cancers. PMID- 15801153 TI - Preparation of chromosomes from plant leaf meristems for karyotype analysis and in situ hybridization. AB - A reliable method for preparing metaphase chromosomes from plant leaf tissues is described. The chromosomes are suitable for karyotype analysis and gene mapping by fluorescence in situ hybridisation (FISH). The method is based on enzymatic digestion of young leaf tissues (shoot-tips) after which the resulting protoplasts are treated hypotonically before being dropped onto microscopic slides. Compared to root-tip chromosomes, leaf chromosomes tend to be longer, or less condensed, and hence more karyotypically differentiated. Metaphase index in young leaf tissues is also very high. Metaphase spread consists of evenly and well-distributed chromosomes and this allows accurate counting. The plant used to demonstrate this method is birch (Betula L.), a group of tree species that has extremely small chromosomes. Root-tip chromosomes of these plants are difficult to obtain, as cutting does not produce roots readily. Seedling chromosomes do not represent the same genomic constitution as their mother trees due to introgressive hybridisation. Furthermore, sample collection in the field is convenient and actively growing leaf buds are available throughout the growing season. FISH experiments with these leaf chromosomes also give good results comparable to those obtained with root-tip chromosomes or even better as mapping on long or extended chromosomes has high resolution in general. Mapping of the 16S-28S ribosomal genes on birch leaf chromosomes has been shown to differentiate between birch species and therefore can accurately confirm their interspecific hybrids. PMID- 15801154 TI - An in vitro method for analysis of chondrogenesis in limb mesenchyme from individual transgenic (hdf) embryos. AB - The present study describes a simple, rapid protocol for culture for limb tissue from individual 10.5-day post coitum mouse embryos that supports cartilage differentiation over a six-day period. This technique differs from other commonly used methods utilizing pooled limb tissue in that: 1) forelimbs from individual embryos were used as donor tissue; 2) limb tissue was dissociated by very gentle enzymatic digest (0.1% trypsin, 5 min); and, 3) cell suspensions were plated at a lower density (1 x 10(7) vs. 2 x 10(7) cells/ml) in a reduced volume of 3-5 microl. Under these modified conditions to increase limb cell yield from each embryo, histochemical and immunohistochemical analyses demonstrated reproducible formation of precartilage aggregates and subsequent overt chondrogenesis over a predictable time course. Using this culture protocol, analysis of limb mesenchyme from heterozygous hdf embryos, which bear an insertional mutation of the Cspg2 gene encoding the core protein of the chondroitin sulfate proteoglycan, versican, revealed an overall similar chondrogenic potential to that observed for wild-type littermates. This technique readily enables in vitro culture of limb bud mesenchyme from individual mouse embryos at this developmental stage and may be utilized by investigators to study the effects of the hdf and other transgenic mutations on mammalian limb development in vitro. PMID- 15801155 TI - Development and characterisation of pilchard (Sardinops sagax neopilchardus) cell lines derived from liver and heart tissues. AB - Two cell lines have been established from juvenile pilchards (Sardinops sagax neopilchardus) caught in waters off the Victorian coast of Australia. Following establishment of primary cultures derived from different pilchard tissues, using various cell culture media, a pilchard liver (PL) cell line and a pilchard heart (PH) cell line have been maintained in Eagle's minimal essential medium supplemented with 10% foetal bovine serum for over four years. The cell lines have been cryopreserved in liquid nitrogen and can be recovered from storage with good cell viability. Stock cell cultures have been maintained at 20-22 degrees C on a continuous basis in normal atmosphere (100% air), with weekly subculture at a split ratio of 3:1. The origin of the cell cultures was confirmed by PCR analysis using primers designed to be specific for pilchard mitochondrial DNA. In addition, the liver cell line was cloned and both the parental cell line and clones thereof were shown to be susceptible to a broad range of marine and freshwater viral pathogens of fish. PMID- 15801156 TI - Fluorescence microplate assay for the detection of oxidative burst products in tobacco cell suspensions using 2',7'-dichlorofluorescein. AB - The technique of 2',7'-dihydrodichlorofluorescin diacetate (H2DCF-DA)-derived fluorescence was applied to measurements of the oxidative burst reaction in plant cell suspension cultures in an automatic fluorometric multiwell microplate assay. The developed procedure was found to be versatile and effective for the determination of the oxidative burst reaction in plant cell cultures. Using this assay, cumulative production of reactive oxygen intermediates may be monitored and recorded non-destructively on a real-time scale for a large number of samples at frequent and continual time intervals for time course experiments. Through the use of various inhibitors and inducers or elicitors of the oxidative burst in combination with H2DCF-DA, this assay aids in the dissection of the signal transduction pathways and the determination of the origins of the oxidative burst in plant cells. PMID- 15801158 TI - Sterile polystyrene culture dishes induce transformation of polyps into medusae in Aurelia aurita (Scyphozoa, Cnidaria). AB - Polyps of Aurelia aurita produce medusae once a year. Under laboratory conditions this process, termed strobilation, can be induced by lowering the incubation temperature for about two weeks. Here we report a fast induction of strobilation by sterile polystyrene culture dishes. The effect is abolished when the culture dishes are washed twice with hot water prior to the experiment. We recommend that polystyrene cultureware should be pre-washed whenever there is an indication of unusual effects. PMID- 15801157 TI - Intracellular Ca2+ measurements in live cells by rapid line scan confocal microscopy: simplified calibration methodology. AB - Altered intracellular Ca2+ dynamics are characteristically observed in cardiomyocytes from failing hearts. Studies of Ca2+ handling in myocytes predominantly use Fluo-3 AM, a visible light excitable Ca2+ chelating fluorescent dye in conjunction with rapid line-scanning confocal microscopy. However, Fluo-3 AM does not allow for traditional ratiometric determination of intracellular Ca2+ concentration and has required use of mathematic correction factors with values obtained from separate procedures to convert Fluo-3 AM fluorescence to appropriate CA2+ concentrations. This study describes methodology to directly measure intracellular Ca2+ levels using inactivated, Fluo-3 AM loaded cardiomyocytes equilibrated with Ca2+ concentration standards. Titration of Ca2+ concentration exhibits a linear relationship to increasing Fluo-3 AM fluorescence intensity. Images obtained from individual myocyte confocal scans were recorded, average pixel intensity values were calculated, and a plot is generated relating the average pixel intensity to known Ca2+ concentrations. These standard plots can be used to convert transient Ca2+ fluorescence obtained with experimental cells to Ca2+ concentrations by linear regression analysis. Standards are determined on the same microscope used for acquisition of unknown Ca2+ concentrations, simplifying data interpretaton and assuring accuracy of conversion values. This procedure eliminates additional equipment, ratiometric imaging, and mathematic correction factors and should be used to investigators requiring a straightforward method for measuring Ca2+ concentrations in live cells using Ca2+-chelating dyes exhibiting variable fluorescence intensity. PMID- 15801159 TI - Epithelial cell cultures from Botryllus schlosseri palleal buds: accomplishments and challenges. AB - This study focuses on recent improvement in epithelial monolayer cultures originating from whole extirpated Botryllus schlosseri (Urochordata) buds. Buds (n = 2,000) were taken at different ('A' to 'D') blastogenic stages. We tested the suitability of 35 combinations of various substrates and media on attachment, cell spread, epithelial growth frequencies and on monolayer lifespans. Under favorable conditions, cultured buds at blastogenic stages 'B' to 'D' (but not stage 'A') started to attach to the substrates following a 3-day transient period that leads to formation of spheres and attached monolayers. Substrate type is important for the attachment and the development of monolayers. Under various culture conditions, some of stages 'B' and 'C' buds develop (3-20 days) one or more large (1 mm diameter) spheres. Stage 'D' buds develop monolayers (up to 20% of buds) without going through a sphere phase. Neither spheres nor attached monolayers of epithelium were observed in stage 'A' bud cultures. Spheres grew at a rate of 60 microm in diameter per day using specific medium types and did not attach unless the appropriate substrate was present. When attached, epithelial monolayers expanded at a rate of 200 microm in diameter per day, for 3-15 days, and subsequently detached and died. Sixteen types of media were tested. Medium and substrate combinations were found to determine epithelial lifespan. These results revealed significant improvements in the culture of epithelial monolayers from Botryllus palleal buds. However, an early senescence of the developed epithelial sheets (up to two weeks from onset of appearance) may indicate an internal ageing clock that should be taken into consideration in future approaches. PMID- 15801160 TI - A reproducible modified method for direct preparation of chorionic villi cytogenetic analysis. AB - Chorionic villi are finger like projections, which surrounds the embryonic sac in early pregnancy and will later form the placenta. The outer layer of the chorionic villi consists of trophoblast cells. These cells are actively dividing and can be used for direct preparation or short-term culture for prenatal diagnosis of chromosomal abnormalities. We herein describe a 100% reproducible modified technique for one day direct culturing (direct preparation) of trophoblast cells. Direct preparation of chorionic villi provides the physician with additional diagnostic information during prenatal diagnosis and genetic counseling. Direct preparation also eliminates the risk of maternal cell contamination. This method was 100% reproducible provided all the precautions mentioned herewith were taken into consideration and can be completed in 45 minutes upon arrival of the samples in the laboratories. PMID- 15801161 TI - Establishment, characterization and viral susceptibility of 3 new cell lines from snakehead, Channa striatus (Blooch). AB - Three cell lines were established from muscle (SHMS), heart (SHHT) and swim bladder (SHSB) of snakehead (Channa striatus). The cells grew initially at 25 degrees C in L15 medium supplemented with 20% fetal bovine serum and have been subcultured 13-18 times since their initiation on June 25, 2002. Growth of the snakehead cells was serum-dependent and plating efficiencies ranged from 22-29%. These snakehead cells grew well in RPMI 1640 and L-15 media, which are commonly used for cultivation of animal and mammalian cells and retained 95.9-96.6% cell viability following storage for 4 months in liquid nitrogen. Karyotyping indicated that these snakehead-derived cell lines remained diploid with a chromosome count of 44 at their early passage (passage 8-14). These cell lines were sensitive to CCV, VHSV, SVCV, IPN and SHRV; they were refractory to IHNV. These newly established cell lines are currently being used for the investigation of snakehead viral diseases in Hawaii and will be available for future isolation and study of snakehead viruses. PMID- 15801162 TI - A new method to obtain epithelial and stromal explants from human Corneo-Scleral Discs for the routine culture of corneal epithelial and fibroblast cells. AB - Acquisition of human corneal cells for culture is hindered not only by the scarcity of donor tissues but also by some of the standard enzymatic and mechanical isolation techniques. Good yields have been reported from full thickness explant and sclero-limbal pieces. However, due to their greater proliferative capacity, fibroblasts will encroach and subsequently overwhelm epithelial cultures whichever technique is used. The novel approach presented here is to minimise this by removal of the whole stroma from the epithelial layers at the outset. This is achieved by selective sectioning with the Webb mini microtome developed in the Norwich Eye Research Laboratory. The microtome can be sterilised by alcohol spraying or autoclaving and is small enough to use in the culture hood. A selective cut in the region of the Bowman's membrane results in the isolation of the epithelium from the stroma and thus exposed, the basal epithelial layers are released from contact inhibition to allow growth. The stroma is further cut to produce multiple sections for the culture of fibroblasts. Both pure epithelial and stromal fibroblast cultures have been successfully generated in serum-enriched medium as well as defined serum-free media with growth supplements, from the corneo-scleral discs of donors of all ages. PMID- 15801163 TI - Isolation and primary culture of gill and digestive gland cells from the common mussel Mytilus edulis. AB - As the marine mussel Mytilus edulis is commonly used as a sentinel species, it would be useful to develop a primary culture of the target organs most often in contact with the marine environment. This study reports an improved method for dissociating the digestive gland and gills of M. edulis and considers the effect of mussel storage on cell viability and functionality before culture initiation. Viability and enzymatic activities such as those of esterase and peroxidase were monitored by flow cytometry, a sensitive, objective technique allowing large volumes of cells to be counted within a short time. A primary culture of digestive gland showed more than 75% viability after 72 h. Mussels were maintained in an aquarium containing clean, oxygenated seawater at 12 degrees C for two days before culture initiation, and dissociation was performed mechanically and chemically with Ca-Mg-free saline to obtain digestive gland cells. Application of non-specific esterase activity, using fluorescein diacetate (FDA test) coupled with flow cytometry, characterised the functionality of digestive gland and gill cells in culture. PMID- 15801164 TI - Analysis of cytosolic and lysosomal pH in apoptotic cells by flow cytometry. AB - Several reports indicate that the cytosol is acidified during apoptosis although the mechanism is not yet fully elucidated. The most acidic organelle found in the cell is the lysosome, raising the possibility that lysosomal proton release may contribute to the cytosolic acidification. We here describe methods for measurement of the cytosolic and lysosomal pH in U937 cells by a dual-emission ratiometric technique suitable for flow cytometry. Cytosolic pH was analysed in cells loaded with the fluorescent probe BCECF, while lysosomal pH was determined after endocytosis of FITC-dextran. Standard curves were obtained by incubating cells in buffers with different pH in the presence of the proton ionophore nigericin. Apoptosis was induced by exposure of cells to 10 ng/ml TNF-alpha for 4 h, and apoptotic cells were identified using a fluorescent marker for active caspases. By gating of control and apoptotic cells, the cytosolic and lysosomal pH were calculated in each population. The cytosolic pH was found to decrease from 7.2+/-0.1 to 5.8+/-0.1 and the lysosomal increased from 4.3+/-0.4 to 5.2+/ 0.3. These methods will be useful in future attempts to evaluate the involvement of lysosomes in the acidification of the cytosol during apoptosis. PMID- 15801165 TI - Data reproducibility in fluorescence image analysis. AB - Fluorescence image analysis provides quantitative data on fluorescence in situ hybridization signals (FISH), immunofluorescence labelings, Green Fluorescent Protein (GFP) expression and microarrays. It is a valuable tool for decision making in the fields of biology and medicine. The aim of this study was to evaluate the reproducibility of fluorescence intensity measurements and standardization when acquisitions are performed under various but well defined conditions. Fluorescent intensity of standard beads (Inspeck series, Molecular Probes) was repeatedly measured using an image analyzer and automated procedures. Images were acquired using several integration times and neutral filter sets. A standardization procedure was used for expressing the data in a same unit: data were multiplied by the light attenuation factor and were divided by the CCD integration times. Results show that 1) standardization is possible 2) accurate and reliable fluorescence measurements can be obtained and 3) specimens showing large differences in fluorescence intensity can be objectively compared. Moreover fluorescent test slides including fluorochrome solutions and altuglas slides were tested for shading correction and as overall test systems. PMID- 15801166 TI - Cobblestone area measuring (CAM) assay: a new way of assessing the potential of human haemopoietic stem cells. AB - A new in vitro way of scoring the potential (proliferative- differentiative) of human haemopoietic stem cells (HHSC) is presented. We have applied it in the investigation of HHSC behaviour in normal bone marrow specimens under culture conditions. This system describes a modification of some existing culture assays for primitive HHSC function, and proliferation of the cobblestone area forming cell (CAFC) was determined by counting the cobblestone areas (CA) produced every week. The main steps of the assay are: (a) culture of MNC fraction on pre established fibroblastic confluent layers from the M2.10B4 mouse cell line; (b) weekly counting of CA over a period of 5 weeks; (c) weekly in situ observation of CA morphology; (d) enumeration of secondary progenitors per CA, in the 5th week. The CAM index, expressing the value of the CA kinetic line slope and the number of secondary progenitors per CA were evaluated as factors indicating HHSC proliferative and differentiative potential, respectively. This culture system has the potential to serve as an in vitro assay of human stem cell transplantation. It could be applied to evaluating the potential of HHSC contained in haemopoietic collections of diverse origin, only in combination with measurements of the starting number of CAFC. PMID- 15801167 TI - Establishment, growth, cryopreservation and species of origin identification of three cell lines from white sturgeon, Acipenser transmontanus. AB - Three cell lines derived from fin (WSF), head soft tissue (WSHST) and body muscle (WSBM) were established from white sturgeon (Acipenser transmontanus). Characterization included determination of optimal growth kinetics, karyotyping, and mitochondrial ribosomal RNA (rRNA) genotyping. The primary cultures of these cells were generated by the explant technique using the L-15 medium supplemented with 20% fetal bovine serum and epidermal/fibroblast growth factors. The cells grew between 15-30 degrees C, but optimal growth occurred at 25 degrees C with a doubling time of 48 hours. The cell lines can be readily maintained in-vitro and have been subcultured over 35 times. Following cryopreservation in liquid nitrogen, thawed cells exhibit a viability of > 90% after a 16-month storage period. Chromosomal typing of these cell lines at their 17th passage revealed a chromosomal distribution of 242 to 278 with an apparent peak ranging from 250 to 260. Polymerase chain reaction amplification of mitochondrial 16S ribosomal RNA and sequence analysis indicated 100% identity of the sequences found in the cell lines with those found in the source animal, confirming that the cell lines were of A. transmontanus origin. PMID- 15801168 TI - Coulter counter use in the enumeration of muscle and fat stem cells. AB - Although the manual counting chamber (hemacytometer) is the gold standard for counting cells, this method is subject to great variability due to the 'human factor'. The automated cell counter (Coulter Counter) can enumerate cells in less time and with greater accuracy than the hemacytometer by removing many of the steps in which errors are made. While the Coulter Counter (and others of its type) has been used for many years in the cell culture field, there have been few studies to validate its use with specific cell types. We conducted several experiments in which we assessed the accuracy of the Coulter Counter over counts made with a hemacytometer as well as validated its use for the counting of satellite cells and preadipocytes. PMID- 15801169 TI - Short-term serum supplementation improves glucose-oxygen deprivation in primary cortical cultures grown under serum-free conditions. AB - Brain ischemia can be studied in vitro by depriving primary neurons of oxygen and glucose by replacing oxygen with argon and glucose with its antimetabolite 2 deoxy-D-glucose. In this contribution, we explain how to construct a reliably functioning ischemia chamber and use it to study neuronal cell death in neuron enriched fetal primary cortical cultures grown under serum-free conditions. We observed that these cultures exhibited a significant cell death even during exposure to oxygenated balanced salt solution used as control for oxygen-glucose deprivation. We show that addition of only 2% fetal calf serum 24 h prior, during, and after treatment almost abolished this undesirable cell loss and proportionally increased cell death induced by oxygen-glucose deprivation. Western blots and immunocytochemistry showed that these effects were mainly due to an increase in neuronal viability under control conditions accompanied by a limited glial proliferation independent of the treatment condition. Under these modified conditions, the cultures could also still be effectively preconditioned by a short-term oxygen-glucose deprivation. In summary, this modified protocol combines the advantages of serum-free neuronal culture, where potentially toxic antimitotic substances can be omitted, with a serum-mediated protection of neurons against unspecific factors and concomitant sensitization for oxygen glucose deprivation. PMID- 15801170 TI - A rapid and reliable flow cytometric method for determining Hsp70 levels in tobacco protoplasts. AB - Current methods to determine heat shock protein (Hsp) synthesis or accumulation in plant cells, such as Western blotting and biometabolic labelling are either indirectly quantitative, labour-intensive or biohazardous. An optimal flow cytometric protocol was developed to measure the intracellular Hsp70/Hsc70 levels in tobacco protoplasts. After heat treatments, protoplasts were fixed in 2% paraformaldehyde-phosphate-buffered saline and dehydrated overnight in methyl cellusolve, followed by permeabilization with Triton X-100 (0.1% in Protoplast Wash Fluid). Immunolabelling of Hsp70/Hsc70 was done for 1 hour with a mouse monoclonal antibody and detected by R-Phycoerythrin-conjugated goat anti-mouse IgG using flow cytometry. Flow cytometry detected a significant 1.2-fold increase in Hsp70/Hsc70 accumulation (P < 0.001) in protoplasts, while Western blotting, quantified by image analysis, showed induction under similar conditions but at lower significance (P < 0.05). The coefficients of variance for flow cytometry and Western blotting were 30.7 and 49.8 respectively. Optimum temperature of heat induced Hsp70/Hsc70 accumulation in tobacco protoplasts occurred at 40 degrees C. Flow cytometry is proposed as a quantitative, more reproducible and rapid alternative to Western blotting for the detection of Hsp70 accumulation in plant cells. PMID- 15801171 TI - Measurement of hemoglobin in long-term fixed erythroid cells: application development for cell science experiments in microgravity. AB - Studying the effects of microgravity on cell differentiation will enhance our understanding of fundamental biology and is indispensable for a sustained space program. Rauscher murine erythroleukemic cells were chosen as a model system to study erythroid cell differentiation aboard the International Space Station because these cells undergo differentiation in response to the natural inducer, erythropoietin, as well as various chemical-inducers. We have now developed a method to quantify hemoglobin in Rauscher cells after weeks of fixation and storage required for such space biology experiments. By exploiting the pseudoperoxidase activity of hemoglobin and by using reagents that yield a soluble chromophore that freely passes out of fixed cells, we developed a highly specific and sensitive assay applicable to cells fixed as long as 4 months. PMID- 15801172 TI - The status of hospital discharge data in Denmark, Scotland, West Germany, and the United States. AB - This section has surveyed characteristics of the health service systems in Denmark, Scotland, West Germany, and the United States. Certain characteristics in each country are likely to increase hospital utilization. In the United States the characteristics include economic incentives to physicians and patients for hospital use and the high percent of physicians in specialty practices. In Denmark the high proportion of hospital-based physicians may increase hospital use. In Scotland the use of capitation to pay office-based physicians and the absence of a nursing home system probably increase hospital use, and in West Germany the large number of physicians per population and the lack of sufficient alternative facilities for long-term care are likely to increase hospital use. On the other hand, each health services system has some characteristics that should decrease hospital use: for instance, the large number of alternative facilities for long-term care in the United States, the well-established programs for home care in Denmark and Scotland, and the small percent of physicians who are specialists in West Germany. Further research is needed to understand the interactions of these factors and the effects they have on hospital use. Increased understanding of the effects of health services system characteristics should result in more useful comparisons of hospital utilization statistics. PMID- 15801173 TI - The influence of the ionic medium on the behaviour of sodium glycocholate and cholate aqueous solutions. AB - In the framework of research on the sodium bile salts, solutions of sodium cholate and glycocholate had been studied only at three ionic medium concentrations. In this paper, experiments in two other concentrations of ionic medium are carried out both to verify the behaviour dependence on the ionic medium and to apply the results of this paper to a future investigation on solutions containing contemporary sodium cholate and glycocholate. To this aim, solubility, protonation constants, lead (II) salts solubility products at 25 degrees C and in 0.300 and 0.750 mol dm(-3) N(CH3)4Cl were determined. The formed species in solutions containing lead (II) together with the selected bile salts and the behaviour of the sodium salts in micellar and premicellar solutions were investigated, too. The obtained results were compared with those obtained at the other ionic medium concentrations. PMID- 15801174 TI - PM-10 and heavy metals in particulate matter of the province of Lecce (Apulia, Southern Italy). AB - This paper shows the results of a preliminary study of air monitoring in the province of Lecce (Apulia, Southern Italy). In particular, the attention has been focused on the determination of the PM-10 level and of the concentration of nine metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb, V and Zn) present on the filters which were collected in two towns (Lecce and Campi Salentina) from 2002 until 2003. The metals have been chosen on the basis of their toxicity and of their possible use as chemical tracers. The results have proved that PM-10 values and metals concentrations did not show substantial difference between the two towns and that PM-10 level and lead concentration are below the limit established by Italian law in force. The experiments have demonstrated a high correlation between iron and manganese in both sampling sites; this could be attributed to pollution of metallurgical origin. Multivariate statistical analysis, carried out by HCA and PCA methods, has been used in order to differentiate samples in relation to sampling sites, sampling period and meteorological conditions. PMID- 15801175 TI - Analysis of pellets found in the north area of the Asiago plateau (Vicenza, Italy), a locality of the 1914-1918 World War. AB - Pellets of unknown material contained in an aluminium cylinder were found in the north area of the Asiago plateau (Vicenza), a locality of the First World War (1914-1918). Elemental analysis, infrared, chromatography and NMR experiments indicate that the main product is pentaerythritol tetranitrate (PETN). This substance was probably an igniting primer used by Austro-Hungarian (A.U.) military engineering. Hypothesis of medical use of the pellets can be disregarded while it seems improbable the use of this explosive by German Army during the Second War World. PMID- 15801176 TI - Multivariate optimization of an axially-viewed inductively coupled plasma multichannel-based emission spectrometer for the analysis of environmental samples. AB - An experimental design procedure was applied to optimize the operating conditions of an axially-viewed inductively coupled plasma emission spectrometer instruments equipped with echelle optics with cross dispersion and charge transfer device. The multivariate effect of carrier gas flow rate and r.f. power on several analytical figures was investigated and discussed. Both ultrasonic and pneumatic nebulization were used. For the final choice of the optimum, different criteria were taken into account, mainly plasma robustness, instrumental precision, analyte and background net emission, detection limits and signal-to-background ratios. It was found that the use of moderate power (1100W) and mean carrier gas flow rate (0.75 L/min) allows to obtain sufficient plasma robustness, satisfactory precision, and excellent signal-to-background ratios and limits of detection, favorable for ultratrace element determinations in environmental matrices. PMID- 15801177 TI - Azorhodanine derivatives as inhibitors for acidic corrosion of nickel. AB - Azorhodanine derivatives (HL1-HL5) were tested as corrosion inhibitors for nickel in 2M HNO3 solution using weight loss and galvanostatic polarization techniques. The results showed that these derivatives act as inhibitors for nickel in this medium. The inhibition was assumed to occur via adsorption of the inhibitor molecule on the metal surface. Polarization measurements indicated that these compounds act as mixed-type inhibitors, but the cathode is more polarized when an external current was applied. This means that these compounds retard the rate of hydrogen evolution and the rate of dissolution of the metal. Results showed that azorhodanine derivatives are adsorbed on the nickel surface following Temkin's adsorption isotherm. The activation energy and thermodynamic parameters were calculated and discussed at different temperatures (30-45 degrees C). PMID- 15801178 TI - Simultaneous determination of phenol and mononitrophenol isomers using PLS regression and conventional and derivative spectrophotometry. AB - The partial least squares regression method (PLS) was tested as a calibration procedure for the simultaneous determination of phenol, o-nitrophenol, m nitrophenol and p-nitrophenol by both conventional and first derivative UV/Vis spectrophotometry. The experiments were conducted in the acidic, neutral and basic media. The results obtained by the application of the PLS procedure on the conventional and first derivative spectra in two solvent media were compared. It was found that the results obtained in the basic medium have better performance characteristics than those obtained in the acidic or neutral media. Comparable results were obtained in the case of both conventional and first derivative absorbance data. The proposed method was applied to the determination of the four phenol derivatives in natural spiked water samples at concentration levels between 1.0 and 10.0 microg ml(-1) with average recoveries in the range 96% - 99%. PMID- 15801179 TI - Determination of trace cadmium in waters by flame atomic absorption spectrophotometry after preconcentration with 1-nitroso-2-naphthol-3,6-disulfonic acid on Ambersorb 572. AB - A procedure for the determination of trace amount of cadmium after adsorption of its 1-nitroso-2-naphthol-3,6-disulfonic acid chelate on Ambersorb 572 has been proposed. This chelate is adsorbed on the adsorbent in the pH range 3-8 from large volumes of aqueous solution of water samples with a preconcentration factor of 200. After being sorbed, cadmium was eluted by 5 mL of 2.0 mol L(-1) nitric acid solution and determined directly by flame atomic absorption spectrophotometery (FAAS). The detection limit (3sigma) of cadmium was 0.32 microg L(-1). The precision of the proposed procedure, calculated as the relative standard deviation of recovery in sample solution (100 mL) containing 5 microg of cadmium was satisfactory (1.9%). The adsorption of cadmium onto adsorbent can formally be described by a Langmuir equation with a maximum adsorption capacity of 19.6 mg g(-1) and a binding constant of 6.5 x 10(-3) L mg(-1). Various parameters, such as the effect of pH and the interference of a number of metal ions on the determination of cadmium, have been studied in detail to optimize the conditions for the preconcentration and determination of cadmium in water samples. This procedure was applied to the determination of cadmium in tap and river water samples. PMID- 15801180 TI - Kinetic spectrophotometric determination of rutin by its inhibitory effect on the oxidation of amaranth by potassium periodate. AB - A novel kinetic spectrophotometric method is described for the determination of rutin. The method is based on the inhibitory effect of rutin on the oxidation reaction of amaranth by potassium periodate in acidic media at 100 degrees C. The linear range for the determination of rutin is 0.02 - 0.50 microg/ml, and the detection limit is 0.014 microg/ml. The method has been successfully applied to the determination of rutin in medicine of rutin tablet and traditional Chinese medicine. PMID- 15801181 TI - Physicochemical profile studies of soil along the banks of Ubu river in Ekwusigo and Nnewi local government areas of Anambra State of Nigeria. AB - A physicochemical profile study on the soil along the banks of the Ubu river that runs across Ekwusigo and Nnewi Local Government Areas (LGA) of Anambra State of Nigeria was investigated for possible evidence of industrial waste disposal by the various large to medium scale industrial activities taking place within the two LGAs. When compared with soil samples from Uzoakwa Ihiala (pH 5.44) with little or no industrial activities, the soil along the banks of the course of the river were found to be more acidic with mean pH values that are lower than 4.7. Similarity in mean sulphate (7.01 mg/kg), phosphate (12.95 mg/kg) and chloride (242 mg/kg) contents was observed between the soils from the banks at Nnewi with those of the Uzoakwa Ihiala that was used as control. This was attributed to contributions from domestic sources rather than from industrial sources, and partly to the similarity of their soil chemistry that may be indigenous to both areas. Electrical conductivity values (55 -74 microS/cm) show that the ionic content of the soil is lower than those of the control (129 microS/cm). This may have been explained by the higher extractible Ca (175.73 mg/kg), K (34.32 mg/kg), and Mg (1.94 mg/kg) recorded for the control. The abundance of the metals extractible from the soils of the river banks follow the sequence Ca>K>Na>Fe. Results for the digested soil samples show that Fe is the most abundant metal in the soil with mean values that are close to 13,000 mg/kg, but is much lower than the mean value of 39,210 mg/kg recorded for the control. Both bank and control samples can be characterized as ferrous. The relative abundance of metal ions observed in the digested soil samples from the river banks follow the sequence Fe>>Al>Ca>Na>K>Zn>Cr. Values of 4,565 mg/kg and 300 mg/kg were recorded for total hydrocarbon content and lead respectively for soils around Cento Group, indicating that there were industrial waste disposal or spillages on soil around the area, which was not observed in other areas. PMID- 15801182 TI - Buffers for the physiological pH range: acidic dissociation constants of zwitterionic compounds in various hydroorganic media. AB - The measurement of pH of a physiological buffer is an integral part of clinical diagnosis. For the standardization of pH and control of acidity in the physiological region of pH 7 to 9, as found in blood and plasma. Good et al., Ferguson et al., and Bellini et al. have listed hydrogen buffers which are N substituted amino acids compatible with common biological media. Recently, the acid base behavior of BICINE, in aqueous solution and in different solvent mixtures has been studied by us. Other investigators reported similar results for (CAPS and CAPSO), (BES), (ACES and CHES), TRICINE, TES, ADA, (HEPPS and MOPSO), (MOPS), and (MOBS and HEPBS). These studies have provided precise knowledge about the suitability of a biological buffer. No work seems to have been done on the determination of second stage dissociation constants of 2-(4 Morphlinomethyl)propenoic acid, 2-[bis(2-hydroxyethyl)amino methyl]propenoic acid, 2-[bis(2-hydroxypropyl)aminomethyl]propenoic acid, and 2-[N-ethyl-N-(2 hydroxyethyl)aminomethyl]propenoic acid in aquo-organic solvent mixtures. PMID- 15801183 TI - Synthesis and efficiency of a spherical macroporous epoxy-polyamide chelating resin for preconcentrating and separating trace noble metal ions. AB - The determination of noble metals in various materials usually requires their preconcentration and separation from other elements. In spite of the improvements in analytical instrumentation and the development of new analytical techniques such as ICP-MS, which are capable of detecting metal ions at ppt levels, the interference caused by the sample matrix still exists and is perhaps the most serious problem, making a pre-determination enrichment step necessary. Thus, the search for efficient preconcentration and separation methods is essential. A series of chelating resins that can selectively adsorb noble metal ions from aqueous solutions have been described. Functional groups, such as salicylaldoxime and thiosemicarbazide have been incorporated in cross-linked polymers or porous silica gel. These resins have very high selectivity for one or several types of noble metal ion. However, desorption of noble metals from these resins is usually difficult. Hence, the development of an adsorbent from which noble metals can be easily desorbed is needed. In this paper, a new spherical macroporous epoxy polyamide chelating resin that met this requirement was synthesized by one step reaction. The synthesis of the resin was safe, rapid and more simple and economical than many report adsorbents. Meanwhile, the resin showed more advantages: better acid and alkali resistance; higher adsorption capacity and lower preconcentration concentrations. A resin column procedure combined with inductively coupled plasma atomic emission spectrometry (ICP-AES) for the determination of trace Rh(III), Ru(III) and Ir(IV) in real samples was established. PMID- 15801184 TI - Is there a double standard when it comes to dementia care? AB - Dementia, a diagnostic syndrome that includes Alzheimer's disease (AD) and vascular dementia, affects more than 4 million Europeans. Its prevalence will increase as the population ages, even though half of cases remain undiagnosed. Dementia has a marked impact on the general health economy. The direct costs of caring for persons with AD alone exceed the cost of heart disease, cancer and stroke combined, primarily because of the need for expensive care in the later stages of the disease. Yet research spending for AD remains disproportionately small. Effective planning for the public health issues posed by AD mandates reduction of either the dependency or the prevalence of the disease. Some drug treatments have alleviated the symptoms of AD and improved the quality of life for patients and their families. However, many countries have introduced inappropriately stringent regulations for the use of medical therapy. These and related issues were addressed during the recent Facing Dementia Forum, in which 175 experts in dementia care discussed results of an extensive international survey. The experts identified three main barriers to effective care: (i) dementia is not considered a health care priority; (ii) there is a mistaken belief that nothing can be done about dementia; and (iii) early dementia is difficult to recognise. These observations provided a basis for the Agenda for Change, a series of strategies to overcome these barriers. The specific findings of the survey, as well as the Agenda for Change, will be reviewed in subsequent articles in this supplement. PMID- 15801185 TI - Inequalities in dementia care across Europe: key findings of the Facing Dementia Survey. AB - The Facing Dementia Survey was undertaken to assess the awareness of and behaviours surrounding Alzheimer's disease (AD) and dementia among all key stakeholders in Europe. Interviews were conducted with more than 2500 persons, including caregivers, members of the general population, physicians, persons with AD and influencers of health care policy. Four key messages emerged from the results. First, AD often remains undiagnosed until symptoms become moderate or severe. This delay may be because of the difficulty of recognising the symptoms of early AD and the attribution of symptoms to so-called normal ageing, the fear of AD common among older people, inadequate screening tools for use by physicians and/or a delay in the confirmation of the diagnosis once suspicion is raised. Second, a majority of respondents perceive their governments as indifferent to the economic, social and treatment burdens associated with AD. Third, a substantial majority of caregivers, physicians and the general population appreciate the wide-ranging impact that AD can have on the quality of life of people who suffer from it and their informal caregivers. While most caregivers reported life-changing negative effects, a few also noted some positive aspects to their experience. Finally, survey results revealed that most caregivers and members of the general public do not have sufficient information about the benefits of treatment and care. The findings of the Facing Dementia Survey support the development of an agenda for change in dementia care across Europe. PMID- 15801186 TI - A caregiver's perspective: the family's life is changed forever. PMID- 15801187 TI - Implications of the Facing Dementia Survey for the general population, patients and caregivers across Europe. AB - The Facing Dementia Survey included interviews with approximately 1200 members of the general public, 600 persons who served as primary caregivers for a person with Alzheimer's disease (AD) and 96 persons with AD. Members of the general public and the caregivers indicated level of agreement on a 5-point scale to a series of attitudinal statements about the disease as well as other diagnostic questions. Persons with AD underwent a qualitative interview. Results indicate that Europeans are acutely aware of the potentially devastating consequences of AD, but few are familiar with the early-stage symptoms. Most members of the general population accept that early intervention and therapy are key to slowing the effects of the disease. Caregivers often delayed consulting a physician about a loved one's behaviour, partly because of uncertainty about early signs of AD. They also expressed doubt about the ability of physicians to make the diagnosis. Caregivers experience profound effects of AD in every area of their life- emotional, physical, social and financial. Persons with AD responded to their diagnosis in one of three ways: a fatalistic attitude, a belief that their problems are linked to old age or a lack of acceptance. Survey respondents from three populations were critical of the level of government investment in and support of AD. PMID- 15801188 TI - A caregiver's perspective: how we kept Kirsten's spirit alive. PMID- 15801189 TI - Implications of the Facing Dementia Survey for health care professionals across Europe. AB - Approximately 600 general practitioners, primary care physicians and specialists in six European nations (France, Germany, Italy, Poland, Spain and UK) who treat patients with Alzheimer's disease (AD) were interviewed during the Facing Dementia Survey. Compared with generalists, specialists displayed the most optimism regarding the effects of age, believing that health and memory do not inevitably deteriorate as one grows older. Most physician respondents agreed that the diagnosis of AD is too often delayed. A primary reason cited for this delay was the difficulty experienced by both physicians and the general public in identifying early signs of AD. Many physicians believed treatments are available that can slow the disease course. The vast majority surveyed in each nation believed that early treatment of AD can delay disease progression [mean, 87%; range, 68% (United Kingdom) to 96% (Poland)]. More than half of physicians who initiate treatment in France (66%), Germany (59%), Italy (82%), Poland (82%) and Spain (69%) said they institute treatment for AD immediately after diagnosis. The exception was the United Kingdom, where 48% initiated treatment immediately, whereas more than half waited at least a month to start therapy. To a large extent, physicians saw the governments of their countries as a hindrance rather than a help in caring for persons with AD. PMID- 15801190 TI - A caregiver's perspective: fighting to make dementia patients more visible. PMID- 15801191 TI - Implications of the Facing Dementia Survey for policy makers and third-party organisations across Europe. AB - The Facing Dementia Survey included qualitative interviews conducted with 60 persons who influence health care policies in six European countries (France, Germany, Italy, Poland, Spain and the United Kingdom). Respondents generally reinforced the perception of other survey participants that governments do not adequately fund research into the causes and cure of Alzheimer's disease (AD) and dementia. (AD is the most common cause of dementia, and throughout this discussion, issues raised in reference to AD apply also to dementia in general.) Access to care services and treatment remains highly restricted in some countries and may vary even within countries. Yet, AD presents an enormous but unappreciated social and economic burden for a growing segment of society. AD is associated with unique challenges to health care systems, because diagnosis is difficult and the disease has an enormous impact on the affected persons and their caregivers alike. Moreover, there is a lack of trained professionals to diagnose and manage the disease, a lack of human and financial resources to provide care and services for people with AD as well as their families and a lack of infrastructure to deliver needed services. These difficulties are compounded by inadequate education of both the general public and physicians. Pressure to change current governmental policy towards neurodegenerative diseases may come from the increasing prevalence of AD as the population ages and from broader awareness of the total societal costs of long-term care. PMID- 15801192 TI - A patient's and a caregiver's perspective: James' diagnosis was not easy for our family. PMID- 15801193 TI - An agenda for change for dementia care across Europe. PMID- 15801194 TI - [In the foreground of fundamental investigations. The 50th anniversary of "Fiziologichnyi Zhurnal"]. AB - 50 years have passed from time of publication of the first number of "Fiziologichnyi Zhurnal", that is the scientific-theoretical edition of a National Academy of Sciences of Ukraine and O.O. Bogomolets Institute of Physiology. "Fiziologichnyi Zhurnal" is a leading Ukrainian academical periodical edition, informing scientific community about a condition of physiology and pathophysiology, state-of-the-practice in physiology. In different years the journal was headed by the outstanding scientists--physiologists academicians G.V. Folbort, A.F. Makarchenko, F.N. Serkov. Now the editor-in-chief of "Fiziologichnyi Zhurnal" is corresponding member NAS of Ukraine V.F. Sagach. PMID- 15801195 TI - [Attenuation of acetylcholine-induced hyperpolarization of aortic endothelial cells in rats fed by high-salt diet and its correction with enalapril]. AB - High-salt diet is known to result in attenuation of endothelium-dependent relaxation in rat aorta. We investigated the possible alterations in electrical responses of rat aortic endothelial cells to acetylcholine as one of the eventual mechanisms that lead to inhibition of endothelium-dependent relaxation induced by high-salt diet. It was shown that endothelial resting membrane potential in rats fed by high-salt diet did not differ significantly from that observed in the control group (-40.6+/-2.3 mV and -41.2+/-0.9 mV, respectively). The amplitude of acetylcholine-induced hyperpolarization of endothelial cells in rats fed by high salt diet was significantly attenuated (9.0+/-1.5 mV vs 22.7+/-1.3 mV in the control group). The amplitude of hyperpolarization increased to 25.7+/-1.3 mV when high-salt diet was administered with enalapril, the angiotensin-converting enzyme inhibitor. We conclude that high-salt diet leads to attenuation of hyperpolarization of rat aortic endothelial cells in response to acetylcholine, that may mediate inhibition of NO synthesis and endothelium-dependent relaxation. Angiotensin-converting enzyme inhibitor enalapril prevents the observed alterations in endothelial electrical responses induced by high-salt diet. PMID- 15801196 TI - [Dynamic interaction between plasma membrane structures and intracellular calcium stores in neurons of the spinal cord ganglia]. AB - We studied the dynamic contribution of endoplasmic reticulum and mitochondria to depolarization-induced Ca2+ transients in small (18-24 microm) DRG neurons of rats. We have used the application of 10 microM of mitochondrial protonophore CCCP for switching off the calcium uptake by mitochondrial uniporter. For depletion of the store of endoplasmic reticulum we applied 1 microM of thapsigargin. Depolarization-induced transients in control conditions and in conditions when one of the mechanisms (mitochondria or endoplasmic reticulum) does not participate in the forming of the shape of Ca2+ transient have been studied. This allowed us to clarify the kinetics of mitochondrial and endoplasmic reticulum uptake and release of calcium in the process of the neuronal activity. We have determined the main characteristics of functioning of above-named calcium stores in the process of cell excitation, such as time of the beginning of uptake, time and duration of maximum activity etc. We concluded, that mitochondria and endoplasmic reticulum are acting in opposite directions at least in the phase of the beginning of the transient. Mitochondria are limiting the amplitude of the transient during depolarization, at the same time the endoplasmic reticulum is increasing the amplitude of the transient by CICR (calcium-induced calcium release) mechanism. Mitochondria store calcium released from endoplasmic reticulum by application of 30 mM caffeine. Inhibition of the mitochondrial uniporter results in reduction of amplitude of repetitive caffeine application compared with control conditions. We have compared the kinetics of mitochondrial participation in the formation of calcium signal when the initial sources of calcium ions were different. Our results allow us to suggest a close functional dynamic interactions between mitochondria and endoplasmic reticulum during calcium signaling in sensory neurons. PMID- 15801197 TI - [The effects of omega-3 polyunsaturated fatty acids on lipid peroxidation]. AB - The aim of this study was to evaluate the effects of a diet supplemented with omega-3 polyunsaturated fatty acids (PUFAs) (plant-derived alpha-linolenic acid and eicosapentaenoic and docosahexaenoic acids with alpha-tocopherol (product "Tekom") on the myocardial phospholipid fatty acids composition, lipid peroxidation and activity antioxidant enzymes--superoxide dismutase and catalase in myocardial tissue in control and after ischemia/reperfusion of isolated working rat hearts. Inclusion of omega-3 PUFAs into the animals' diet within 4 weeks demonstrated that alpha-linolenic acid and "Tekom" increased omega-3 polyunsaturated fatty acids content in cardiomyocytes membranes and limited lipid peroxidation (decreased content of congugates of dienes and malone dialdehyde, reduced hemiluminescence of myocardial tissue). Additionally omega-3 PUFAs caused the beneficial effects on activity of antioxidant enzymes in cardiac tissue (increased superoxide dismutase and catalase activity). PMID- 15801198 TI - [Effect of nitric oxide on the efficiency of oxygen consumption by the working skeletal muscle in fatigue]. AB - The influence of NO on the efficiency of oxygen usage by a skeletal muscle under fatigue of dog's gastrocnemius muscle was investigated. In control experiments was shown, that 10 short-term (30") electrical stimulation (8 Hz, 5 ms, 20 V) with 5" interval resulted in significant reduction of the muscle contraction force (more than 40%) and increased considerably oxygen cost of muscle gastrocnemius work (more than 130%) compared to the initial parameters. The registered depression of the muscle contraction force testified to development of gastrocnemius muscle fatigue, accompanied by mitochondrial factor (MF) appearance in blood from femoralis vein, which, as shown by us earlier, is a marker of the mPTP opening. Injection of L-NMMA, a NOS inhibitor (2.7 mg/kg, i.a.) resulted in pronounced fall (more than 1.5 times) of the initial force parameters, in comparison with the control experiments. Under these conditions the magnitude of oxygen cost of gastrocnemius muscle work exceeded control parameters considerably. The development of gastrocnemius muscle fatigue under L-NMMA action was accompanied, as well as in the control condition by the mPTP opening. The preliminary injection of sodium nitroprusside, a NO donor (0.2 mg/kg, i.v.) prevented a fall of muscle contractions force and considerable inhibition of oxygen usage efficiency by gastrocnemius muscle under conditions similar to control. Furthemore, gastrocnemius muscle fatigue was not developed, and MF concentration in blood from femoralis vein was much lower, than in the control experiments, that testified to absence of the mPTP opening. Apparently, preliminary short-term (30") electrical stimulation (8 Hz, 5 ms, 20 V) with 2' interval, created the precondition effect and raised the level of authentic NO. Under these conditions, as well as under preliminary injection of the NO donor, we did not register the marked inhibition of oxygen usage efficiency and development of gastrocnemius muscle fatigue. At the same time, MF in blood from v. femoralis was practically absent, that testify to absence of the mPTP opening. Thus, NO in physiological concentration by inhibition of mPTP opening, can prevent decrease of oxygen usage efficiency and development of the working skeletal muscle fatigue. PMID- 15801200 TI - [Adenosine triphosphate-dependence of volume sensitive chloride current in LNCaP cell line of human prostate cancer]. AB - Although sensitivity to intracellular ATP is considered to be one of the hallmarks of swelling activated Cl- current (I(Cl,swell)) involved in regulatory volume decrease (RVD) following hypotonic stress, the type and manner of such sensitivity seems to vary in different cell types. Here by using whole-cell patch clamp recording we investigated ATP sensitivity of I(Cl,swell) in LNCaP human prostate cancer cell line. Suppression of endogenous ATP production with metabolic inhibitors (oligomycin, iodoacetate and rotenone) during cell dialysis with ATP- and Mg2+-free pipette solution did not prevent I(Cl,swell) in response to hypotonic exposure. However, supplementing this solution with 5 mM Na-ATP led to the development of I(Cl,swell) with nearly 305 higher density and less pronounced voltage-dependent inactivation (manifested mainly by the increase of non-inactivated current component) at positive potentials. On the contrary, inclusion of 1 mM Mg2+ in the patch pipette resulted in even smaller I(Cl,swell) (30% lower density compared to Mg2+-free conditions), which inactivated completely on prolonged depolarization. The presence of 5 mM Mg-ATP in the pipette did not affect I(Cl,swell) density. Neither intervention significantly altered the rate of I(Cl,swell) development in response to hypotonicity. We conclude that intracellular ATP, a positive modulator of I(Cl,swell)-carrying volume-regulated anion channel (VRAC) in LNCaP cells most likely acts via binding rather than hydrolysis and/or phosphorylation reactions, whereas intracellular Mg2+ is VRAC inhibitor. PMID- 15801199 TI - [Role of nitric oxide in effects of intramedullary injected gamma-aminobutyric acid on blood circulation]. AB - In acute experiments on anaesthetized with urethane normotensive rats, we studied the influence of modulation of neuronal NO-synthase (nNOS) activity on the effects of GABA injected in the populations of cardiovascular neurons within the nucleus of tractus solitarius (NTS), n.ambiguus (AMB), paramedian reticular nucleus (PMn) and lateral reticular nucleus (LRN). The data obtained give evidence for nNOS involvement in the effects of intramedullary injected GABA on the cardiovascular system. PMID- 15801201 TI - [Thapsigargin-sensitive and insensitive intracellular calcium stores in acinar cells of the submandibular salivary gland in rats]. AB - Acinar cells of rat submandibular salivary gland are characterized by heterogeneity of intracellular Ca2+ stores. In the present work we have studied this heterogeneity using Arsenazo III dye to measure a cellular total calcium content and Fura-2/AM, to determine free cytosolic calcium concentration ([Ca2+]i). We have found that the amount of Ca2+ released by inhibition of Ca2+ ATPase of the ER with thapsigargin comprises approximately 30% of total ER calcium. This result was obtained in experiments on both intact and permeabilized acinar cells. We have also shown that both Ca2+ ATPase inhibition with thapsigargin and emptying the stores with acetylcholine (ACh) led to activation of store-operated Ca2+ influx (an increase in total calcium content of approximately 14%). In permeabilized cells application of ACh after preincubation with thapsigargin led to a further decrease in total cellular calcium content (approximately 38%). At the same time in intact cells it resulted in generation of [Ca2+]i transients with gradually decreasing amplitudes. Thus, ACh is capable of producing an additional release of Ca2+ from thapsigargin-insensitive stores. This additional release is IP3-dependent since it was completely blocked by heparin. We conclude that in acinar cells of rat submandibular gland thapsigargin sensitive and thapsigargin-insensitive Ca2+ stores could exist. PMID- 15801202 TI - [Allelic polymorphism of endothelial NO-synthase (T(-786)-->C) promoter gene as risk factor of acute coronary syndrome]. AB - Frequency of promoter endothelial NO-synthase gene allelic polymorphism by using polymerase chain reaction and restriction fragment length polymorphism (RFLP-PCR) was determined in 221 patients with acute coronary syndrome (ACS) and in 83 almost healthy subjects. Data obtained indicate that different promoter allelic variant frequency differs significantly in patients with ACS and in control group. Correlation of normal homozygotes (T/T), heterozygotes (T/C) and pathologic homozygotes (C/C) was 48%, 36% and 16% respectively in patients, and in control it was 48%, 46%, 6% (P<0.05 by c2-test). Thus, in patients with ACS in Ukrainian population pathologic C/C variants of 5'-flanking region of eNOS gene were found 2.7-times more often in ACS patients, than in control. This allows us to suggest, that this allelic polymorphism can be considered as one of genetic risk factors of ACS development. PMID- 15801203 TI - [Age-related characteristics of the thymus and adrenal cortex function in CBA mice immunized by T-dependent antigen]. AB - The thymic serum factor (FTS) titer and corticosterone level in blood of adult and old CBA mice after immunization by T-dependent antigen was investigated. It has been revealed that in adult mice these indices influenced by phase changes in dynamics of immunization whereas in old mice fluctuations of FFS and corticosterone levels were monotonous. In both adult and old mice the correlation exists between thymic hormone and corticosterone. The increasing of thymic function by thymaline administration did not improve the dynamics of FTS titer and corticosterone level in immunized old mice. PMID- 15801204 TI - [Effect of constant darkness on circadian rhythm of the kidney excretory function in white rats]. AB - In experiments on White rats was established, that the steady-state darkness during 8 days results in desynchronizes of the circadian rhythms of sodium ions, titrate acids and ammonium excretion with moving from 23.00-2.00 hours to 2.00 14.00 hours. PMID- 15801205 TI - [Blood circulation and secretory function in the gastric mucosa during inhibition of M1-cholinergic receptors and L-Ca2+-channels]. AB - The gastric secretion and gastric mucosal blood flow (GMBF) under simultaneous action of M1-cholinergic antagonist gastrocepine (3 mg/kg i.v.) and L-Ca2+ channels blocker verapamile (1.25 mg/kg i.v.) were investigated in urethane anaesthetized rats. The stomach was perfused with saline, and GMBF was measured by using hydrogen gas clearance technique. The acid production under combined action of both blockers was lower by 57.3% and 18.6%, than in case of separate action of gastrocepine and verapamile accordingly. Pepsin concentration decreased by 32%, which did not differ from verapamile action alone. The above changes of gastric secretion was accompanied by significant decrease of GMBF. Inhibition of M1-cholinergic receptors by gastrocepine did not show their key role in basal gastric secretion in rats, but they exacerbate L-Ca2+-channels blocker action. We conclude, that enhance of acid output inhibition is related to the simultaneous depression of nervous impulses in ganglionic neurons and acethylcholine release from parasympathetic postganglionic neurons. Synchronous block of M1-cholinergic receptors and L-Ca2+-channels of smooth muscles stomach vessels resulted in reducing of gastric mucosal blood flow. PMID- 15801206 TI - [The model of hypokinesia in rats and preventing its negative effects]. AB - The special container for modeling of long-term hypokinesia state in rats was developed. Such construction allows us to examine simultaneously hypokinesia and its negative influence correction by gas mixture regulated with different oxygen partial pressure. PMID- 15801207 TI - [Role of genetic factors in the development of osteoporosis]. AB - Osteoporosis which often accompanies the normal aging and a number of pathological conditions is closely related to bone remodeling disturbances. This process is provided mainly by the genesis of osteoblasts and osteoclasts, their activities and interactions. All the above in turn is dependent on numerous systemic and local regulatory factors such as hormones, nutrients, minerals, vitamins, cytokines, their receptors etc. Recently the crucial role of genetic polymorphism of many receptors in bone remodeling, bone loss and susceptibility to osteoporosis development has been shown. This review is devoted to the genetic background of interrelationships among the hormones, cytokines, vitamins and their receptors and discussion of possible sources of discrepancies in data obtained by many authors who studied populations like genetic, diet, life-styles and environmental influences. PMID- 15801208 TI - [Adenine triphosphate-sensitive potassium channels: classification, structure and functions]. AB - The review is dedicated to classification, structure and function of ATP sensitive potassium channels. A general classification of K+ channels into families is based upon the primary amino acid sequence of the pore-forming subunits. The earliest classification of potassium channels was made according to the main mechanism of activation or opening of the channels. Activation of the voltage-sensitive channels is regulated by changes in the membrane potential, and of the ligand-sensitive channels by a number of ligands (calcium ions, ATP, neurotransmitters and G-protein). PMID- 15801209 TI - [A new rapid method for indication of tick-borne encephalitis viral antigen]. AB - A coagglutinizing reagent has been designed, which makes it possible to make a rapid diagnosis of tick-borne encephalitis in the coagglutination test within 2-5 minutes. Laboratory tests of the agent have shown that the coagglutination test is sensitive and specific (it is many times more sensitive than indirect hemagglutination test and 2-4 times more sensitive than enzyme immunoassay). The trials conducted on natural and clinical materials also have confirmed the sensitivity and specificity of this method. The complex use of the coagglutination test and enzyme immunoassay to examine ticks whether they contain tick-borne encephalitis viral antigen allows their diagnostic effectiveness to be enhanced by 2.4 times. The clinical trial has indicated that the coagglutination test may be useful in detecting the antigen of tick-borne encephalitis virus in the blood of infected and sick patients in acute and chronic forms. PMID- 15801210 TI - [The clinical features of the acute period of mixed infection of tick-borne encephalitis and Ixodes tick-borne borreliosis in children]. AB - The paper presents the results of a study of 176 cases of mixed infection of tick borne encephalitis and Ixodes tick-borne borreliosis in children in the Kuznetsk coal fields. The mixed infection presented as meningeal, meningoencephalitic, and feverish forms. Mixed infection is characterized by a severe course and it is not attended by the symptoms that are characteristic for monoinfections, which requires the immunological verification of their diagnosis. Unlike tick-borne encephalitis, the meningeal and meningoencephalitic forms of the mixed infection are marked by the great magnitude and duration of a temperature reaction and overall cerebral and focal manifestations. The outcome is characterized by the formation of symptoms of chronic tick-borne encephalitis in 25% of the cases of the meningoencephalitic form of the mixed infection. PMID- 15801211 TI - [Algid malaria]. AB - Algid is a rare complication of tropical malaria and it occurs in 0.37% of cases. Algid malaria is characterized by hemodynamic disorders as shock with pronounced metabolic changes and hypothermia. A number of factors are involved in the development of algid malaria. These include: 1. Pathological phenomena that are associated with the changes in the state of red blood cells and lead to impaired microcirculation (cytoadherence, sequestration, rosetting); 2. Tumor necrosis factor (TNF) that provokes hypoglycemia, coagulopathy, and impaired erythropoiesis; 3. Altered acid-alkali balance with the development of metabolic acidosis; 4. Gastrointestinal lesion. Adherence of contaminated red blood cells in the intestinal mucosal vessels induces epithelial ischemic damage. Impaired absorption of liquid and its loss with vomiting and diarrhea result in acute hypovolemia; 5. Algid malaria is associated with the addition of gram-negative septicemia. The paper describes a case of algid malaria. PMID- 15801212 TI - [The ecological and epidemiological principles of prevention of ascariasis under the conditions of large-scale solid waste storage]. AB - The paper presents the results of a study of the impact of large-scale solid waste storage on ascariasis morbidity in the population. The use of sewage sediments as an organic soil fertilizer to grow strawberries and table greens is shown to substantially increase the risk of Ascaris infection in the population. Storage of solid domestic garbage on specialized dumping grounds does not lead to mass environmental pollution with geohelminthic eggs. PMID- 15801213 TI - [Organization of the epidemiological surveillance system for parasitic diseases in the Republic of Armenia under reorganization of a State Sanitary and Epidemiological surveillance]. PMID- 15801215 TI - [Health and the incidence of enterobiasis and ascariasis in children in the Sverdlovsk Region of Perm]. PMID- 15801214 TI - [Trichinosis in the Republic of North Ossetia-Alania]. AB - The present paper presents the biological, epidemiological, clinical, and immunological characteristics of trichinosis in the Republic of North Ossetia Alania, which is based on the results of long-term studies and follow-ups and on the 1971-2002 report data from medical and veterinary services and hunt inspectorate agencies. It has established that a total of 675 clinical cases of trichinosis were recorded in Ossetia-Alania in the period under study. The incidence rate of the disease ranged from 0.5 to 16.9 cases per 100,000 individuals, which was 5-20 times or more than the average federal rate. The source of infection of human trichinosis was meat from pigs in 70.7% of the cases and that from wild animals (boars and bears) in 21.3%. Immunological examinations (red cell-linked-antigen test and enzyme immunoassay) of patients with trichinosis who had ingested infected meat, but had not fallen ill with its manifestative form revealed 81.8 and 44.2% seropositive subjects, respectively. In different periods, a positive response ranged from 3.6 to 4.7% in healthy urban residents and from 6.4 to 11.8% in rural ones. The extensiveness of invasion was 0.06 to 1.4% in pigs from individual farms and 0.0033 to 0.034% in those from collective farms. In the Digorsk Region, the trichinosis infection rates were 13.4% in dogs, 4.8% in cats, and 1.3% in murine rodents. The extensiveness of invasion in wild boars was 6.35%. A population of one capsule forming type of Trichinella spiralis was shown to circulate in the animals of natural and synanthropic biocenoses and it is just this population that determines the specific features of an epizootic process in this area. The natural and social factors that contribute to the formation and maintenance of the activity of endemic foci of trichinosis in the Ossetia-Alania under the present socioeconomic conditions are analyzed. PMID- 15801216 TI - [Malaria as a re-emerging disease in the countries of the WHO European Region: lessons of history and the present-day situation. Communication 2. The Middle Asia]. PMID- 15801217 TI - [Malaria mosquitoes in Moscow and in the Moscow Region: cytogenetic analysis]. AB - The species and karyotypical composition of malaria mosquitoes in Moscow and in the Moscow Region have been examined. A cytogenetic analysis in the Moscow Region has revealed 3 types of malaria mosquitoes: Anopheles claviger Mg., An. maculipennis Mg., and An. messeae Fall. An. messeae predominates in the Solnechnogorsk and Noginsk Regions, An. maculipennis prevails in the Ramensk and Serpukhov Regions. In the combined habitats, the ratio of two predominant types has been ascertained to vary during a season. The proportion of An. maculipennis increases by the end of summer and decreases during hibernation. The more northern type of An. messeae has been shown to be superior to An. maculipennis in fertility. Thus, the increased relative size of An. maculipennis during summer is due to the higher viability of the larvae of this type. In the Moscow water reservoirs, the larvae of both types develop concurrently; the proportion of An. messeae is 89.8%. The high level of chromosomal variability was found in the urban populations of An. messeae by the inversions of XL1, XL3, 2R1, 3R1. A role of chromosomal variability in the adaptation of mosquitoes to urban conditions is discussed. PMID- 15801218 TI - [Synanthropic fleas (Siphonaptera) biting people in Moscow and control of their size]. AB - The species composition of fleas attacking man in Moscow, the time course of changes in their size, and the efficiency of disinfecting measures against them have been studied. The basement premises are the commonest habitat of fleas. The absolute predominant species in these biotopes are Cetenocephalides felis (Boushe) whose feeders are dogs, synanthropic rodents, and humans, other than cats. Micropopulations of the fleas have been existed in some structures for more than 5 years (a follow-up period). A mathematical analysis of the factors that influence the efficiency of disinfestation has established that the most influential factors were the factors associated with the poor state of basement premises (the presence of a substrate for the breeding and shelter of fleas, the possibility of homeless animals and rodents, the feeders of fleas, to penetrate there) that reduce the residual effect of insecticides. Out of 15 applied insecticides, those based on chlorpyrifos, cypermethrin, and fenvalerate showed the highest efficacy. The combined use of liquid and powered insecticides is most effective in disinfestation. The treatments made in the periods when the number of fleas is small in the basements (April-May and August-September) provide the most prolonged residual effect. PMID- 15801219 TI - [Experimental rationale for the optimization of chemotherapy for alveococciasis]. PMID- 15801220 TI - [Investigation of the trichinocidal activity of the agent VIG-03]. PMID- 15801221 TI - [Method for catching helminthic eggs and protozoan cysts from domestic-drinking water supply sources]. PMID- 15801222 TI - [Outbreak of hemorrhagic fever with renal syndrome in the Ulyanovsk Region]. PMID- 15801223 TI - [On registration of a case of mixed infection: malaria-HIV/AIDS in Minsk]. PMID- 15801224 TI - [Dirofilariasis in the Autonomous Republic of Crimea]. AB - The paper presents data on the prevalence of dirofilariasis in the Autonomous Republic of Crimea during 5 years from 1999 to 2003. The nematode Dirofilaria repens is a causative agent of the disease. Sporadic cases of helminthiasis are annually registered in some urban and rural areas on the plain of the peninsula. The disease is absent on the foothills and Black sea coast of the southern and eastern parts of Crimea. The localization of the parasite is typical of this helminthiasis. Dirofilariasis in Crimea may be regarded as a transmissible helminthiasis with natural focal trends. PMID- 15801225 TI - [Prevalence of toxoplasmosis and methods for its laboratory diagnosis]. AB - The review presents data on the prevalence of toxoplasmosis in different countries and regions. It gives a comparative assessment of the methods of serodiagnosis of toxoplasmosis, which permit identification of specific immunoglobulins G, M, A, and E. The authors show a role of laboratory diagnostic techniques in the differentiation of the acute form of toxoplasmosis from its chronic form. They provide the currently available modifications of the basic procedure for enzyme immunoassay, molecular genetic methods, and a direct fluorescent antibody test for the detection of Toxoplasma gondii antigens. The current view of a cellular immune response and its role in monitoring the development of toxoplasmosis are considered. PMID- 15801226 TI - [Human fascioliasis in South America]. PMID- 15801227 TI - [Cerebral malaria: pathogenesis, clinical picture and treatment]. PMID- 15801228 TI - [Genodiagnosis of cancer: current status and prospects]. PMID- 15801229 TI - [Response of cerebral hemodynamics to combined stress impacts]. AB - Changes in cerebral hemodynamics (CH) was assessed with transcranial color duplex scanning in the middle cerebral artery (MSA) of 15 healthy male volunteers aged 20-23 years in response to psychoemotional stress (PES). The latter was induced by calculations, mild exercise (Martine's test), hypercapnia in inspiration of 6% CO2 and their various combinations. The results showed that a combined action of the stressors differs quantitatively and qualitatively from their isolated action. As a rule, in combination the stressors potentiated each other. The response of CH to a combined action of the stressors, PES and hypercapnia, in particular, may be also characterized by a prevalent effect of one of the stressors, in this case by a stronger action of hypercapnia. PMID- 15801230 TI - [Peripheral mechanisms of vagal sinus arrhythmia: a role of typical affiliation of cardiac M-cholinoreceptors]. AB - Salvo impulses on the vagus nerve of a cat with a varying in each cardiocycle interval after ECG P wave led to recurrent variations of ECG interval P-P manifesting as sinus arrhythmia. A chronotropic effect of the vagus nerve experienced moderate changes in dislocation of the vagus stimulus position in the beginning of cardiocycle lasting about 200 ms after ECG P wave and in ECG segment T-P. Distinct (in the range 100-300 ms) variations of the interval ECG P-P lengthening took place in the change of the vagus stimulus generation moment in the narrow zone of cardiocycle of 40-70 ms duration. The zone location corresponded to the end of ECG T wave. Metoctramine or gallamine block of M2 cholinoreceptors reduced maximal lengthening of the interval P-P and severity of cycle changes of P-P interval in arrhythmia and its variation amplitude under changing moment of vagus stimulus generation in the zone of cardiocycle showing marked shifts of a chronotropic effect. M1-cholinoreceptor block with pirenzepin or receptors of M3-subtype in the action of 4-DAMP produced an opposite effect enhancing these parameters. PMID- 15801231 TI - [Age-specific changes of carbohydrate metabolism in rat liver in immobilization stress]. AB - We have studied peculiarities of the main pathways of glucose utilization in the liver of adult and old rats under immobilization stress. Our data indicate that in both adult and old rats stress is accompanied by an increase in glycolysis intensity in the liver. In stressed adult rats stimulation of glycolysis is associated with metabolism changes characteristic of decreased oxidative decarboxylation of pyruvate in the liver. In immobilized old rats glycolysis stimulation took place in maintanence of effective oxidation of pyruvic acid and enhancement of glucose catabolism in pentose phosphate pathway. PMID- 15801232 TI - [Changes in lipid metabolism and hemostasis in intravenous and intrahepatic transplantation of neonatal allogenic hepatocytes in conditions of experimental hypercholesterolemia]. AB - We studied effects of neonatal hepatocytes transplantation (NHT) on lipid metabolism and hemostasis in experimental hypercholesterolemia. A positive NHT influence on lipid metabolism, lipid peroxidation and hemostasis was established. The experimental data evidence for preferential influence of intrahepatic NHT on lipid metabolism and hemostasis compared to intravenous NHT. PMID- 15801233 TI - [Characteristics of the kidney functional state after a single dose of thyroxine in white rats]. AB - A 50 mcg/100 g b.w. single dose of thyroxine was injected to non-inbred white rat males with body mass 140-180 g in conditions of 5% water and osmotic (3% solution of sodium chloride) load, then renal function was studied. This injection reduced creatinine clearance and phosphate excretion, raised nitrate and protein excretion. The kidney of the hyperthyroid animals retains ability to increase creatinine clearance in response to osmotic load. Thus, the above model demonstrates that in the absence of thyroxine renal toxicity it changes homeostatic renal functions both at the vascular-glomerular and tubular levels. PMID- 15801234 TI - [Effect of opiod peptides on free radical oxidation in a bone regenerate after fracture]. AB - Opioid peptides (DSLET and DAGO) stimulating bone tissue regeneration were studied for effects on content of free radical products in regenerate tissue from the region of leg fracture in mice at various terms of reparative osteogenesis. These opioids reduce concentration of malonic dialdehyde and dienic conjugastes in bone for 10 days after fracture. PMID- 15801235 TI - [A new method of sanation and drainage of the abdominal cavity in experimental generalized suppurative peritonitis]. AB - Elimination of microflora and its products from the abdomena, prevention of commissures, abscesses in postoperative peritonitis are essential for the disease outcome. The sack for this purpose made of polyvinilpirrolidone film (PVF) differs from the standard one by aceptic solution which washes the small intestine, eliminates microflora and products via the tube. This diminishes the count of microorganisms in the abdomena, arrests toxicity, activates immune system. PMID- 15801236 TI - [Mechanisms of immunotropic effect of the drug composition Taban-Arshan (Tibet medicine) on lymphocytes of patients with early rheumatoid arthritis and atopic bronchial asthma]. AB - The extract of the Tibetan medicine Taban-Arshan corrects changes of the immune system in inflammatory diseases--rheumatoid arthritis and atopic bronchial asthma. An immunomodulating effect of the drug consists in its ability to suppress hyperactivation of lymphocytes by normalization of expression of basic activation antigens. PMID- 15801237 TI - [Study of neurotrophic effects of yashaltin mud in experimental model of denervation-reinnervation syndrome of shank muscles in mice]. AB - On the experimental model of denervation-reinnervation syndrome of mouse shank muscles neurotrophic effects of jashaltinsky medicinal mud were studied. In experimental mice therapeutic mud has been applied on the small back, tail and rib areas of the body 10 days after nerve crushing. On day 11 after nerve crushing a stable reproduction of a compound nerve action potential with constant amplitude was revealed in repetitive (50 Hz) nerve stimulation. PMID- 15801238 TI - [The role of endorphines in regulation of pain sensitivity]. PMID- 15801239 TI - Eastern red cedar (red savin), Juniperus virginiana, is the most abundant juniper throughout the eastern United States and southeastern Ontario. PMID- 15801240 TI - The rhinitis-asthma link revisited. PMID- 15801241 TI - Health effects of indoor fungi. AB - OBJECTIVE: To review the nontoxic harmful effects that poor indoor air quality caused by fungi can have on health. DATA SOURCES: We searched PubMed for publications related to the various topics discussed in this review, and we relied on our knowledge of the field. STUDY SELECTION: Where more than one publication was relevant, we attempted to identify a consensus of the reports and cited the most relevant articles. Priority was given to randomized controlled trials and expert reports when available, although much of the information herein relates to laboratory research. RESULTS: Actively growing fungal colonies can release volatile substances that have an unpleasant smell, leading to psychological responses in the occupants such as fatigue and nausea. Symptoms that are likely caused by indoor fungi include respiratory complaints that involve the nose and lungs, eye symptoms, and mucous membrane irritation. These adverse effects can occur by a variety of mechanisms, including IgE-mediated hypersensitivity, fungal infection, irritant reaction to spores or fungal metabolites, and possibly toxic reaction to mycotoxins. CONCLUSIONS: Reduced fungal exposure can reasonably be expected to improve health. Removal of moisture from the indoors and proper maintenance of air filters can aid in prevention and elimination of fungi from the home environment. Small areas of present contamination can be cleaned with a dilute bleach solution, which kills viable colonies and removes their mycelia. If fungal contamination is not addressed early, substantial damage can occur, requiring professional remediation. Above all, the individual should not panic at the first sight of fungi growing in the home. Regular inspection and cleaning can prevent many fungus-related problems. PMID- 15801243 TI - Cough and weight loss in a patient with cystic fibrosis. PMID- 15801244 TI - Correlation between nasal symptoms and asthma severity in patients with atopic and nonatopic asthma. AB - BACKGROUND: The association between allergic rhinitis and asthma has been well recognized, and it has been postulated that rhinitis may worsen asthma. OBJECTIVE: To investigate the severity of asthma among patients with atopic and nonatopic asthma with and without nasal symptoms. METHODS: Atopic asthmatic patients and nonatopic asthmatic patients were identified from the records of a university-based asthma clinic. A comparison of demographic clinical features was made within and between these 2 asthmatic groups, dichotomized according to the presence or absence of rhinitis. RESULTS: A total of 178 patients were classified as having atopic asthma and 218 as having nonatopic asthma. The atopic asthmatic patients with nasal symptoms compared with those without had a higher mean forced expiratory volume in 1 second (FEV1), a higher forced vital capacity (FVC), and a higher FEV1/FVC ratio, used fewer oral steroids, and had fewer hospitalizations. The nonatopic asthmatic patients with nasal symptoms compared with those without used more inhaled steroids (and they were also more likely to have nasal polyps on examination). Atopic, relative to nonatopic, asthmatic patients were younger, had a longer duration of asthma, had a higher FEV1/FVC ratio, and took fewer oral steroids. CONCLUSION: Contrary to current hypotheses, in this study the severity of asthma among atopic asthmatic patients was less in those with nasal symptoms. Conversely, among the nonatopic asthmatic patients, asthma was more severe among those with nasal symptoms than those without nasal symptoms. PMID- 15801242 TI - The role of endotoxin and its receptors in allergic disease. AB - OBJECTIVE: To summarize the existing literature on the association of endotoxin with respiratory diseases and allergic sensitization and to review the potentially modifying effects of endotoxin receptor polymorphisms. DATA SOURCES: English-language articles were identified from the MEDLINE and PubMed databases using combinations of the following search terms: endotoxin, toll-like receptor, polymorphisms, atopy, asthma, and allergy. Other sources included experts in the field and the bibliographies of pertinent articles. STUDY SELECTION: Relevant articles were selected based on the authors' expert opinion. RESULTS: Cross sectional studies, particularly those of children raised in rural European communities, suggest that early endotoxin exposure may protect against the development of allergic sensitization and atopic asthma. However, endotoxin exposure may also contribute to other nonatopic respiratory disorders and may exacerbate disease in individuals with preexisting asthma. Paradoxically, among individuals exposed to high levels of endotoxin, carriers of a functional mutation in toll-like receptor 4, which reduces cellular responsiveness to endotoxin, may be at lower risk of developing allergic sensitization. CONCLUSIONS: The effect of endotoxin exposure on allergic sensitization and asthma appears to be influenced by the timing of exposure, the presence or absence of preexisting disease, and polymorphisms in the genes that encode endotoxin receptors. Further studies are needed to define the window period for this effect, as well as the underlying immunologic mechanism. PMID- 15801245 TI - Efficacy and safety of an extended-release formulation of desloratadine and pseudoephedrine vs the individual components in the treatment of seasonal allergic rhinitis. AB - BACKGROUND: Antihistamine-decongestant combination products generally provide more benefit than individual components for adequately treating patients who have seasonal allergic rhinitis (SAR) with moderate-to-severe nasal congestion. OBJECTIVE: To compare the effectiveness and safety of a 24-hour, extended-release formulation of desloratadine and pseudoephedrine with the individual components in patients who have SAR with moderate-to-severe nasal congestion. METHODS: Patients with SAR and significant nasal congestion were enrolled in a multicenter, randomized, double-blind, double-dummy study. Patients were randomly assigned for 2 weeks to once-daily treatment with desloratadine-pseudoephedrine, 5/240-mg tablets; desloratadine, 5 mg; or pseudoephedrine, 240 mg. Primary efficacy variables for the antihistamine and decongestant components of desloratadine-pseudoephedrine were morning and evening reflective total symptom score (TSS), excluding nasal congestion, and morning and evening reflective nasal congestion score during the 2-week treatment period, respectively. Secondary variables included morning instantaneous (end-of-interval) TSS (excluding congestion), nasal congestion score, reflective morning and evening individual symptom scores, overall condition of SAR, and therapeutic response. RESULTS: A total of 1121 patients were enrolled in the study, and 1047 patients completed the 2-week study. Desloratadine-pseudoephedrine was significantly more effective than desloratadine or pseudoephedrine monotherapy in reducing morning and evening reflective TSS (excluding nasal congestion) during the entire treatment period. Desloratadine-pseudoephedrine also was significantly more effective in reducing the morning and evening reflective nasal congestion score compared with desloratadine or pseudoephedrine monotherapy. Significant differences were seen between the desloratadine-pseudoephedrine and monotherapy groups for changes in morning instantaneous TSS (excluding nasal congestion) and nasal congestion scores. No unusual or unexpected adverse events were reported. CONCLUSION: Desloratadine-pseudoephedrine, 5/240-mg once-daily tablets, provided additional benefit over desloratadine, 5 mg, or sustained-release pseudoephedrine, 240 mg, monotherapy in the treatment of patients with SAR and moderate-to-severe nasal congestion. PMID- 15801246 TI - A modified pulmonary index score with predictive value for pediatric asthma exacerbations. AB - BACKGROUND: Several clinical asthma scores have been derived from combinations of physical findings in pediatric asthmatic patients. OBJECTIVE: To test the reproducibility and validity of one such score, the Modified Pulmonary Index Score (MPIS), and to evaluate its predictive value in children hospitalized for asthma. METHODS: In the MPIS, 6 categories are evaluated: oxygen saturation, accessory muscle use, inspiratory to expiratory flow ratio, degree of wheezing, heart rate, and respiratory rate. For each of these 6 measurements or observations, a score of 0 to 3 is assigned. To evaluate the reproducibility of the MPIS, inpatients with status asthmaticus were examined by an attending physician, nurse, and respiratory therapist who were blinded to the other observers' scores. To evaluate the validity of the MPIS as a scale of severity of illness in asthmatic patients, the score at admission was compared with selected outcomes in the same patients. RESULTS: A total of 30 patients participated in this study (mean +/- SD age, 7.6 +/- 5.5 years). Our finding revealed that the MPIS is highly reproducible with a high degree of interrater reliability across caregiver groups (physician to nurse: r = 0.98; 95% confidence interval [CI], >0.96; physician to respiratory therapist: r = 0.95; 95% CI, >0.92; nurse to respiratory therapist: r = 0.94; 95% CI, >0.90). The admission MPIS positively correlated with intensive care unit admission (P < .001), days of continuous albuterol therapy (P = .002), days of supplemental oxygen (P = .002), and length of hospital stay (P = .004). CONCLUSIONS: The MPIS is a highly reproducible and valid indicator of severity of illness in children with asthma. To our knowledge, this is the first pediatric clinical asthma score demonstrated to be reproducible across groups of health care professionals who treat pediatric patients with asthma. PMID- 15801247 TI - Monitoring nebulizer use in children: comparison of electronic and asthma diary data. AB - BACKGROUND: Measurement of nebulizer medication adherence that relies on self report of medication use is subject to recall bias and increased patient burden. Electronic monitoring of nebulizer medication use is relatively new technology and provides an objective measure of nebulizer use. OBJECTIVE: To examine levels of agreement for nebulizer use between self-report on diary cards and electronic monitor data in young inner-city children with asthma. METHODS: Of 221 enrolled children with persistent asthma, 157 (71%) provided 12 weeks of diary card and nebulizer monitor recordings that were matched by date across days and by patient. Concordance, sensitivity, specificity, and kappa coefficients were calculated between self-report and electronic data. RESULTS: The children were predominantly African American (89%) and male (66%), with a mean age of 4.6 years. Their persistent asthma was categorized as mild (61%) or moderate to severe (35%). Concordance between diary and electronic data was 85%, with overreporting on diary cards noted on 15% of the total days. Sensitivity of the diary data relative to the electronic data ranged from 0.80 to 0.91 during the 12 week study. Diary return rates decreased from 75% during the initial 3 weeks to 44% at 12 weeks. CONCLUSIONS: Electronic monitoring of nebulizer use provides a more precise measure of long-term medication use than does self-report on diary cards, and it is feasible for use in high-risk populations. However, diary cards seem to be a valid alternative for short-term monitoring of nebulizer use, resulting in only a slight overestimation of medication use. PMID- 15801248 TI - Exercise-induced dyspnea in children and adolescents: if not asthma then what? AB - BACKGROUND: Exercise-induced dyspnea (EID) in children and adolescents is a common manifestation of asthma and is therefore commonly attributed to exercise induced asthma (EIA) when present in otherwise healthy children. OBJECTIVE: To report the outcome of evaluations for EID when other symptoms and signs of asthma were absent or if there was no response to previous use of an inhaled beta2 agonist. METHODS: We reviewed the results of all exercise tests performed in otherwise healthy patients with EID during 1996 to 2003. Physiologic measures included preexercise and postexercise spirometry with the addition of oxygen uptake, carbon dioxide production, continuous oximetry, and electrocardiogram monitoring during most tests. EIA was diagnosed if symptoms were reproduced in association with a 15% or greater decrease in forced expiratory volume in 1 second from baseline. Endoscopy was performed if stridor and/or decreased maximal inspiratory flow were present. Criteria were established for restrictive abnormalities, physical conditioning, exercise-induced hyperventilation, and normal physiologic limitation. RESULTS: A total of 142 patients met our criteria for inclusion. EID had been present in these patients for a mean duration of 30.2 months (range, <1 to 192 months) before evaluation and had been previously attributed to asthma by the referring physician in 98 of them. Symptoms of EID were reproduced during exercise testing in 117 patients. EIA was identified as the cause of EID in only 11 of those 117. Seventy-four demonstrated only normal physiologic exercise limitation; 48 of these 74 had normal to high cardiovascular conditioning, and 26 had poor conditioning. Other diagnoses associated with reproduced EID included restrictive abnormalities in 15, vocal cord dysfunction in 13, laryngomalacia in 2 (1 of whom had unilateral vocal cord paralysis), primary hyperventilation in 1, and supraventricular tachycardia in 1. CONCLUSION: The diagnosis of EIA should be questioned as the etiology of EID in children and adolescents who have no other clinical manifestations of asthma and who do not respond to pretreatment with a beta2-agonist. Exercise testing that reproduces symptoms while monitoring cardiac and respiratory physiology is then indicated to identify causes of EID other than EIA. PMID- 15801249 TI - Trends in asthma prevalence, hospitalization risk, and inhaled corticosteroid use among alaska native and nonnative medicaid recipients younger than 20 years. AB - BACKGROUND: Few trend data on asthma prevalence exist for U.S. indigenous populations, and none exist for Alaska Natives. OBJECTIVE: To document the epidemiologic features of asthma in Alaska Natives and nonnatives stratified by urban (Anchorage) and rural (non-Anchorage) residence. METHODS: We conducted a retrospective review of Alaskans younger than 20 years enrolled in Medicaid during 1999 to 2002. Asthma was defined as a claim for International Classification of Diseases, Ninth Revision, codes 493.0x to 493.9x plus asthma associated medication during the same calendar year. RESULTS: Among 117,080 Medicaid enrollees, the 4-year asthma prevalence was 3.1% and was 40% to 90% greater for urban residents regardless of race. Yearly prevalence increased from 1.0% to 2.2% (P < .001), with increases in all subgroups. Of 4 predominantly Alaska Native census areas, the area with resident pediatricians and previous participation in asthma research had a 4-year asthma prevalence 5- to 11-fold higher than the other areas. Among persons with asthma, yearly hospitalization risk decreased (from 9.3% to 6.8%; P = .02) concurrent with an increase in the yearly use of inhaled corticosteroids (from 50% to 64%; P < .001). Urban Alaska Natives had the greatest decrease in hospitalization risk and the greatest increase in inhaled corticosteroid use. CONCLUSIONS: Relatively dramatic demographic differences and temporal trends in asthma prevalence occurred in the absence of known differences or changes in risk factor prevalences. This suggests a role for differences in the use of asthma as a diagnosis for respiratory illness. Failure to diagnose and thus treat asthma may affect outcomes because decreases in hospitalization risk were temporally associated with increases in inhaled corticosteroid use. PMID- 15801250 TI - Cardiodepressant and neurologic actions of Solenopsis invicta (imported fire ant) venom alkaloids. AB - BACKGROUND: We hypothesized that the alkaloid compounds that are the majority components of fire ant (Solenopsis invicta) venom are capable of producing cardiovascular and central nervous system toxic effects in mammals. OBJECTIVE: To evaluate toxic effects of synthetic S. invicta alkaloids in rodent models. METHODS: Cardiovascular effects of intravenous injection of the racemic (+/-)-cis and trans-isomers of 2-methyl-6-nundecylpiperidine (isosolenopsin A and solenopsin A, respectively) were evaluated in anesthetized, gallamine-paralyzed rats who had received artificial ventilation and in isolated, perfused rat hearts. RESULTS: (+/-)-Solenopsin A dose dependently (3-30 mg/kg [10 to 104 micromol/kg]) depressed cardiovascular function. Maximal percent changes following injection of 30 mg/kg were -42.96% +/- 5.8% for blood pressure, -29.13% +/- 3.6% for heart rate, and -43.5% +/- 9.2% for left ventricular contractility (dP/dt). (+/-)-Isosolenopsin A (3-15 mg/kg [10 to 52 micromol/kg]) produced responses similar to those seen with the corresponding doses of solenopsin A. In conscious, spontaneously breathing rats, solenopsin A (30 mg/kg intravenously) caused seizures, respiratory arrest, and death. Infusion of working, isolated, perfused hearts with solenopsin A reduced contractile function (dP/dt) at 10 microM and caused cardiac arrest at 100 microM. CONCLUSIONS: Two alkaloid components of imported fire ant venom possess robust cardiorespiratory depressant activity and elicit seizures in the rat. Such effects identify these alkaloids as toxic compounds in biological systems and may explain the cardiorespiratory failure noted in some individuals who experience massive fire ant stings. PMID- 15801251 TI - Natural history of large local and generalized cutaneous reactions to imported fire ant stings in children. AB - BACKGROUND: There are no published data on the natural history of large local and generalized cutaneous reactions to imported fire ant (IFA) stings in children. OBJECTIVE: To determine the natural history of large local and generalized cutaneous reactions to IFA stings in children not treated with immunotherapy by reviewing medical records from a venom clinic during the past 20 years. METHODS: Patients were selected for the study if they were 16 years or younger at the time of the reaction, had only cutaneous symptoms (generalized cutaneous or large local), and did not initiate immunotherapy. Each patient's parents were asked to recall whether the patient had experienced any further stings since the last evaluation at the venom clinic. RESULTS: We contacted 31 of 57 patients evaluated between July 10, 1984, and February 5, 2004. Twenty patients (65%) reported that they had not developed more severe reactions with subsequent stings. Reactions remained cutaneous only. Eleven patients (35%) had not been stung again since the original evaluation. Five of these patients had moved out of the IFA-endemic region. None of the previously evaluated patients reported subsequent life threatening anaphylaxis from IFA stings. CONCLUSIONS: These limited data on IFA stings suggest a benign outcome in children 16 years and younger with large local or generalized cutaneous reactions. Larger and more extensive studies need to be conducted to further define the natural history of cutaneous reactions to IFA stings in children. PMID- 15801252 TI - Squamous cell carcinoma-related antigen in children with acute asthma. AB - BACKGROUND: Increased serum levels of squamous cell carcinoma-related antigen (SCCA) have been observed in patients with allergic disorders, such as atopic dermatitis and bronchial asthma. T(H)2 cytokines, which are known to be involved in the pathogenesis of allergic disorders, stimulate new synthesis of SCCA in cultured human airway epithelial cells. OBJECTIVE: To investigate whether SCCA levels increase during acute exacerbations of asthma in children and whether the T(H)2 cytokines, interleukin 4 (IL-4) and IL-13, are associated with SCCA levels. METHODS: Serum levels of SCCA, IL-4, and IL-13 were measured by enzyme immunoassay during the acute phase of an asthma exacerbation (on hospital admission) and in the recovery phase (after symptoms had subsided). RESULTS: In the 35 children who participated in this study, serum levels of SCCA were significantly elevated in the acute phase (mean +/- SD, 3.09 +/- 2.03 ng/mL) compared with the recovery phase (mean +/- SD, 1.47 +/- 0.64 ng/mL) of an asthma exacerbation (P < .001). In 12 children, the IL-13 levels were observed to correlate with SCCA levels during the recovery phase (r = 0.68, P = .02) but not during the acute phase of an asthma exacerbation. CONCLUSIONS: Serum SCCA levels increase during the acute phase of an asthma exacerbation. During this phase, the increased synthesis of SCCA is not associated with IL-13 but rather mediated by other undefined stimuli. IL-13 may contribute to the basal production of SCCA in asthmatic children. PMID- 15801253 TI - Acrodermatitis enteropathica-like eruption and food allergy. AB - BACKGROUND: Acrodermatitis enteropathica-like eruption (AE) is a distinct rash associated with profound zinc deficiency. It is seen in a variety of conditions but has not been reported as a presentation of food allergy. OBJECTIVE: To report AE as an unusual presentation of food allergy in infants. METHODS: Acrodermatitis enteropathica-like eruption was diagnosed by a characteristic rash and a low serum zinc level. The diagnosis of food allergy was made by history, serum total IgE and food specific IgE levels, or oral challenge with suspected foods. RESULTS: Two infants with AE, diarrhea, and low serum zinc levels were evaluated. Food allergy was found in both infants. The first infant had a serum IgE level of 4642 IU/mL. Specific IgE levels to milk, soybean, wheat, and peanut were 39.04, 10.14, 5.65, and 102.61 kU/L, respectively. Oral challenges to milk and peanut were positive and to soybean were negative. The second infant had a serum IgE level of 991 IU/mL; specific IgE levels to soybean and milk were 36.9 and 0.53 kU/L, respectively. Evaluation for other possible causes of diarrhea revealed homozygous delta F508 in the first infant, confirming the coexistence of cystic fibrosis; findings in the second infant were negative. CONCLUSIONS: Undiagnosed food allergy can lead to profound zinc deficiency. Food allergy should be suspected in a child with acquired AE. PMID- 15801254 TI - Late-onset anaphylaxis to fermented soybeans: the first confirmation of food induced, late-onset anaphylaxis by provocation test. AB - BACKGROUND: Late-onset anaphylactic reactions without early-phase reactions are rarely reported. The hypothesized mechanism of late-onset anaphylaxis to fermented soybeans is delayed absorption or release into the bowel rather than an immunologic phenomenon. OBJECTIVES: To investigate the mechanisms of late-onset anaphylaxis to fermented soybeans in 2 patients and to characterize the allergens involved in anaphylaxis caused by fermented soybeans. METHODS: Two patients underwent skin prick-by-prick tests with fermented soybeans as is. We used an open challenge for the provocation test of anaphylaxis and measured changes in plasma histamine, plasma tryptase, serum eosinophil cationic protein, and plasma leukotriene B4 levels in 1 patient. In addition, specific IgE against fermented soybeans and the allergens of fermented soybeans were detected by enzyme-linked immunosorbent assay and immunoblotting, respectively. RESULTS: The results of the prick-by-prick tests with fermented soybeans as is were positive in both patients and negative in control subjects. The challenge with 50 g of fermented soybeans caused generalized urticaria and dyspnea 13 hours after ingestion of fermented soybeans in 1 patient. In addition, his plasma histamine and tryptase levels transiently elevated during the anaphylactic event. In enzyme-linked immunosorbent assay, the patients showed elevated IgE levels to the proteins of fermented soybeans. Serum IgE antibodies of patients 1 and 2 were bound to approximately 5- and 26-kDa proteins in immunoblotting of fermented soybeans, respectively. CONCLUSION: To our knowledge, this is the first report of late onset anaphylaxis provoked by the challenge test half a day after ingestion of fermented soybeans. PMID- 15801255 TI - Safety of ebastine. PMID- 15801256 TI - Antihistamines and driving ability: evidence from on-the-road driving studies during normal traffic. PMID- 15801257 TI - Combination therapy with a long-acting beta2-agonist and an inhaled corticosteroid as initial maintenance treatment in asthma. PMID- 15801258 TI - Delta-aminolevulinic acid-based photodynamic therapy for acne on the body. AB - Intractable acne on the body was clinically treated with photodynamic therapy (PDT) combining oral administration of delta-aminolevulinic acid (ALA) with polychromatic visible light from a metal halide lamp. The total number of body acne patients was 51. A 10 mg/kg B.W. of ALA was administered to the patients, and the acne lesions were exposed to the light emitted from the metal halide lamp. The wavelengths of the light ranged from 540 to 800 nm. The diameter of the irradiated area and the irradiating distance from the lamp were 100 mm and 130 mm, respectively. Under the irradiation conditions, the total fluence rate and the energy-density uniformity of the light were 69.2 mW/cm2 and 11.5%. The total light energy dose in one session was 60-80 J/cm2 for the body. All patients underwent two sessions of PDT and received no other treatments after PDT or during the follow-up period. The interval between the two PDT sessions was 2-4 weeks. The physician's clinical assessment scores as worsened, unchanged, improved, and markedly improved were 0 (0%), 4 (7.8%), 16 (31.4%), and 31 (60.8%) patients, respectively. Adverse effects of very slight discomfort, burning, and stinging were reported during the irradiation. The blood tests of patients before and after oral administration of ALA demonstrated that no liver dysfunction occurred in this study with ALA dosages within 10 mg/kg B.W. Consequently, orally administered ALA-PDT with a metal halide lamp was considered to be effective for treatment of acne on the body. PMID- 15801259 TI - Bicentre experience in the treatment of severe generalised atopic dermatitis with extracorporeal photochemotherapy. AB - Pilot studies have shown an improvement of atopic dermatitis in approximately 65% of patients during extracorporeal photopheresis (ExP) therapy. The purpose of the present clinical trial was to investigate the response to ExP by controlling clinical and laboratory parameters during short term ExP therapy in patients with severe generalised atopic dermatitis. Thirty-five patients with severe, therapy resistant atopic dermatitis were treated with ExP in an open clinical trial at two week intervals over a period of 6 to 10 cycles. Disease activity was measured before each cycle by SCORAD index together with a standardized protocol for blood samples. ExP led to a significant decrease (p < 0.05) in SCORAD from 74.4 +/- 15.5 before to 36.8 +/- 16.8 after ExP therapy (mean 10 cycles). Approximately 70% (24/33 patients = responder) of patients had a favourable response to ExP requiring at least 6 cycles. The decrease in SCORAD was accompanied by a significant decrease of eosinophil cationic protein (27%), sE-selectin (37%) and sIL-2R (53%) levels in serum (p < 0.05). No significant correlation between a decrease in these levels and values of blood eosinophils or lymphocytes was found (p > 0.05). In comparison to responders, most non-responders were characterised by very high levels of total IgE before and during therapy (p < 0.05). The present clinical trial confirms that short term ExP is an effective treatment for certain patients with severe atopic dermatitis based on anti-inflammatory mechanisms. Total IgE could be a predictor of outcome in ExP treatment. PMID- 15801260 TI - A cross-sectional analysis of quality of life in Japanese acne patients using the Japanese version of Skindex-16. AB - We measured the quality of life of Japanese patients with acne using the Japanese version of Skindex-16, a semantically equivalent and validated translation of the original version produced by Chren et al. A total of 210 acne patients were enrolled in this study. The results showed that patients with acne experienced more severe emotional effects from their skin disease than functional or symptomatic effects. The scores for emotional aspects were related to the severity of the patients' acne following a prescribed scale, but the scores were high even when the patients only had a few comedones. Akaike's information criterion showed that the scores for symptoms were related to the severity of the patients' acne, and the patients' emotions were related to the interval between clinic visits. To improve patients' quality of life, treatment of comedones should be fully respected. In addition, dermatologists should encourage patients to visit clinics regularly to help them improve the emotional aspects of their quality of life. PMID- 15801261 TI - Measurement of the impact of atopic dermatitis on patients' quality of life: a cross-sectional and longitudinal questionnaire study using the Japanese version of Skindex-16. AB - The impact of atopic dermatitis on patients' quality of life was measured using the Japanese version of Skindex-16 in a cross-sectional and longitudinal questionnaire study. One hundred sixty-two adult patients completed Skindex-16 and were followed-up with a standard medical therapy. Three to six months after the initial testing, 135 (83.3%) of the patients again completed Skindex-16 and also answered a general question about whether their skin condition had improved, remained the same, or become worse. The scores of Skindex-16 of 162 patients with atopic dermatitis were significantly higher than those of patients with isolated lesions, particularly in the Symptoms and Emotions scales. Patients with severe atopic dermatitis showed significantly higher scores in the three scales (Symptoms, Emotions, and Functioning), and there was a significant positive correlation between the severity and the 3-scale scores. After the follow-up period, 78 of 135 patients (57.8%) reported that their skin condition had improved. Forty-six patients (34.1%) reported that their skin condition had remained the same, and 11 (8.1%) became worse. Among the patients who said their dermatitis had improved, the scores of Skindex-16 significantly decreased. On the other hand, patients who reported their dermatitis worse showed an increase in the scores. These findings suggest that Skindex-16 responsively measures the disease severity and clinical change in the estimation of the effects of atopic dermatitis on patients' quality of life. This practical and sensitive, skin disease specific, quality-of-life instrument is valuable for assessing patients' outcomes, especially their response to therapy, and is useful to understanding and improving the quality of life of patients suffering with atopic dermatitis. PMID- 15801262 TI - The significance of serum nitric oxide levels in Behcet's disease and recurrent aphthous stomatitis. AB - Behcet's disease (BD) is an inflammatory multisystem disorder characterized by recurrent oral and genital aphthous ulcers, arthritis, uveitis, and thrombophlebitis; it can involve several organs. However, recurrent aphthous stomatitis (RAS) can be seen without a confirmed diagnosis of BD. Moreover, there is no way of predicting whether a patient with RAS will develop BD. Nitric oxide (NO) is a free radical synthesized from L-arginine by one of the family of nitric oxide synthase (NOS) enzymes. Increased production of NO during several inflammatory and infectious processes has been recently postulated. Our aim was to investigate the serum NO levels in patients with active and inactive BD and RAS. Forty-six patients with BD, 30 patients with RAS and 30 healthy controls were enrolled in the study. The patients with BD were separated into two groups: clinically active (n = 24) and inactive (n = 22). A blood sample was collected from all subjects in order to determine their serum NO levels. In patients with active BD, higher serum levels of NO metabolite were found in comparison with patients with inactive BD, in patients with RAS, or healthy controls (p < 0.05). We also found higher serum NO metabolite levels in patients with RAS than in healthy controls (p < 0.05). In patients with inactive BD, statistically significant higher levels of serum NO levels were found in comparison with the control group (p < 0.05). However, we found no statistically significant difference between the patients with inactive BD and RAS, which indicated that inactive BD cannot be distinguished from RAS by serum NO levels. We conclude that serum NO levels may be an important marker for estimating the severity of BD. However, further studies are needed to confirm our findings. PMID- 15801263 TI - The treatment of common warts with infrared coagulation. AB - Treatment of warts can involve medical and surgical methods. Infrared coagulation is a surgical method very rarely used in the treatment of warts. We planned an open-labeled prospective study to research the effect of infrared coagulation in the treatment of common warts, comparing it with electrocoagulation, and discussing its applicability as an alternative therapy regimen. Eighteen patients with common warts were included in this study. There were 49 warts of various sizes (1 mm to 1 cm) in these 18 patients. Twenty-seven warts were treated with infrared coagulation, and 22 warts were treated with electrocoagulation. Sixteen patients were treated with both infrared coagulation and electrocoagulation; two patients were treated with only infrared coagulation. The patients were followed up for six months after treatment. Changes in sizes of warts, healing times, and cure rates were compared by Mann-Whitney U test. The mean healing time was 35.5 +/- 5.7 days with infrared coagulation and 32.9 +/- 4.0 days with electrocoagulation. There was not any significant difference between healing times (p > 0.05). Bacterial infection was seen on seven (31.8%) warts treated with electrocoagulation, but no side effects were seen with infrared coagulation. There was a significant difference between side effect rates in two groups (p < 0.01). Nine (33.3%) of the warts treated with infrared coagulation and seven (31.8%) of the warts treated with electrocoagulation recurred in the six-month follow-up period. The overall cure rates at the end of the follow-up period were 66.7% and 68.2% respectively. There was not any significant difference between the cure rates in the two groups (P > 0.05). We conclude that infrared coagulation is a safe and cheap method and should be kept in mind as an alternative modality for the treatment of common warts. PMID- 15801264 TI - Subungual glomus tumor: clinical manifestations and outcome of surgical treatment. AB - Surgical excision is the treatment of choice for subungual glomus tumor. However, the anatomical location has inherent difficulties. We report the outcomes of surgical treatments for subungual glomus tumor. Sixteen patients, who were seen over an eight-year period (1995-2003) and confirmed as gloums tumor by histopathologic examination were reviewed. The most common subjective symptom was pain induced by contact in 81%. The tumor presented as a discolorated spot or subungual nodule and 38% of tumors were acccompanied with nail dystrophy. All tumors showed discolorated spots or subungual nodules. As shown in the Table 2, the dystrophic nail change was found in 38% of tumors. Differently oriented incisions were made according to the location of tumor, matrix, or bed. The original nail plate was restored in eight patients. Thirteen patients (81%) had cosmetically excellent nail plates, and three patients (19%) had partial distal splits of nail plates. There was no recurrence. Our series suggests that a transungual approach with nail avulsion and an incision selected according to the tumor location can produce an excellent outcome with minimal postoperative complications. Dressing with a trimmed nail plate may also be beneficial in managing the wound and preventing postoperative nail deformity. PMID- 15801265 TI - Immunohistochemical differentiation of extra-ocular sebaceous carcinoma from other skin cancers. AB - We performed an immunohistochemical study using routinely processed formalin fixed and paraffin-embedded tissue specimens from 26 cases of extra-ocular sebaceous carcinoma (EOSC) and eight easily available antibodies. They were polyclonal anti-carcinoembryonic antigen (CEA) antibody, monoclonal anti-CEA antibody, anti-breast carcinoma associated antigen-225 antibody (CU18), anti CA15.3 antibody (CA15.3), anti-CD15 antibody (CD15), anti-breast carcinoma associated antigen antibody (B6.2), anti-gross cystic disease fluid antigen-15 antibody (GCDFP15) and anti-Thomsen-Friedenreich antigen antibody (TFA). Squamous cell carcinoma, porocarcinoma, syringomatous carcinoma, malignant clear cell hidradenoma, apocrine adenocarcinoma, and extramammary Paget's disease with underlying adenocarcinoma were used as controls. EOSC was positive for CU18 and CA15.3 in most cases, and for CD15 in a few cases. Squamous cell carcinoma of the skin was positive for CA15.3 in only one case. Porocarcinoma, syringomatous carcinoma and malignant clear cell hidradenoma were positive for CEA, CU18, CA15.3, and B6.2 in most cases. Apocrine adenocarcinoma and extramammary Paget's disease with underlying adenocarcinoma were positive for CEA, CU18, CD15, GCDFP15, CA15.3, and B6.2 in most cases. TFA was positive not only in EOSC but also in other skin cancers. Immunohistochemical examinations using these seven of eight antibodies except for TFA and routinely processed formalin-fixed and paraffin-embedded tissue specimens are beneficial in differentiating EOSC from other skin cancers. PMID- 15801266 TI - Drug-induced hypersensitivity syndrome due to anticonvulsants in a two-year-old boy. AB - Drug-induced hypersensitivity syndrome (DIHS) usually refers to severe cutaneous drug eruption associated with systemic involvement and potentially fatal outcome. We report a 2-year-old Caucasian boy who developed DIHS due to phenytoin and phenobarbital and who showed extensive internal organ involvement. We are alerting that failure to recognize this drug eruption and discontinue the culprit drug may result in increased severity, greater extent of internal organ involvement, and fatal outcome. The recent research about the influence of human herpesvirus 6 co-infection on the pathogenesis of DIHS is also discussed by the authors in this paper. PMID- 15801267 TI - Unsuccessful treatment of extragenital lichen sclerosus with topical 1% pimecrolimus cream. AB - Lichen sclerosus most commonly affects the anogenital region. Spreading into the extragenital regions is rare, and its course is most commonly asymptomatic. Women have been reported to be affected 6 to 10 times more often than men. The etiology of lichen sclerosus is still unknown. The disease is characterized by ivory-white atrophic plaques, and no treatment ensuring complete recovery is available. T cells are also involved in its pathogenesis. Pimecrolimus is a topical inhibitor of T-cells. In the present paper, we present a male patient with lichen sclerosus located only in extragenital regions and report an unsuccessful outcome of treatment with pimecrolimus 1% cream administered topically twice a day for 16 weeks. PMID- 15801268 TI - Subcutaneous solitary fibrous tumor. AB - We describe a unique case of subcutaneous solitary fibrous tumor (SFT) in a 56 year-old female patient. The patient had been aware of a painless soft mass in her back for ten years. The lesion was surgically excised. Histological examination revealed that the well-defined mass was composed of a proliferation of spindle-shaped fibroblastic cells and polygonal cells embedded in a fibrous matrix corresponding to the so-called "patternless pattern". A prominent pericytomatous pattern (hemangiopericytoma-like structures), focal myxoid changes, and thick hyalinized collagen fibers were also observed. Immunohistochemical stainings for CD34 and bcl-2 were positive in the tumor cells. These features are compatible with SFT. We suggest that SFT should be included in the differential diagnosis of subcutaneous spindle cell tumors. PMID- 15801269 TI - Langerhans cell histiocytosis involving the skin of an elderly woman: a satisfactory remission with oral prednisolone alone. AB - Langerhans cell histiocytosis (LCH) is a disorder characterized by neoplastic proliferation of Langerhans cells that rarely involves the skin in adults. A 74 year-old woman presented with a fourteen year history of eosinophilic granuloma and bone involvement caused by LCH. She had received three combination therapy courses of curettage and radiation since 1987 and had remained free of LCH signs for seven years, after which she started to notice brown nodules on her left leg. Biopsy specimens taken from the lesions showed massive proliferations of large histiocytic cells. Immunoperoxidase stainings for CD1a and S-100 protein were positive. Electron microscopy identified Birbeck granules in the cytoplasm of the atypical Langerhans cells. Treatment with oral prednisolone alone has resulted in the patient remaining in complete remission for 12 months. PMID- 15801270 TI - A case of bullous Sezary syndrome. AB - Sezary syndrome is an aggressive variant of cutaneous T cell lymphoma with poor prognosis and clinically characterized by erythroderma and Sezary cells in the blood. Here we report a case of bullous Sezary syndrome. A seventy-year-old male presented with erythroderma and inguinal lymph node swelling. Histopathological examination showed dermal and epidermal infiltration of atypical lymphocytes and Sezary cells could be detected in peripheral blood samples. He was therefore diagnosed as Sezary syndrome. Four months after the onset, he developed bullae on axillary and inguinal areas, featuring subepidermal blistering with basal cell degeneration and dense infiltration of atypical lymphocytes. Autoimmune bullous diseases were excluded by negative immunofluorescence. Bullous forms of Sezary syndrome are extremely rare although several cases of a bullous variant of mycosis fungoides have been reported. PMID- 15801271 TI - Angiotropic B-cell lymphoma with telangiectasia, accompanied by panniculitic formation. AB - A 67-year-old female presented with diffuse edema, generalized telangiectasia, and indurated skin plaques. Histopathological findings of the skin lesion included CD45+/CD79+/CD20+ large lymphoma cells that filled the vascular lumina and infiltrated the subcutaneous tissues with panniculitic formations. Three cycles of a CHOP (cyclophosphamide, adriamycin, vincristine, and prednisolone) chemotherapy regimen resolved the skin manifestations. After five cycles of CHOP, however, she developed dementia-like symptoms. Three cycles of additional intrathecal chemotherapy (methotrexate, cytarabine and prednisolone) did not improve the neurological symptoms. PMID- 15801273 TI - Interobserver agreement in the use of the ABCD rule for dermoscopy. PMID- 15801272 TI - A case of systemic osteomyelitides due to Mycobacterium avium. AB - A case of multiple osteomyelitides due to Mycobacterium avium (M. avium) infection with osteosclerotic bone lesions is reported. A 67-year-old male had been suffering from persistent fever and back pain since October 1999, and 20.0 2.5 mg prednisolone per day was prescribed for continuous inflammatory symptoms in January 2000. Six months later, computed tomography revealed osteosclerotic lesions in the left femur and thoracic vertebrae, but no skin lesion associated with mastocytosis or internal malignancy was identified. In September of 2002, a dome-shaped, soft subcutaneous tumor developed on the upper sternum. Histopathological findings revealed subcutaneous adipose tissue with several foci of tiny abscesses. Two weeks later, creamy pus was discharged through a draining sinus at the center of the wound. M. avium was demonstrated in the pus by Zeel Nielsen staining and microplate hybridization. PMID- 15801274 TI - Effectiveness of distal protection with the GuardWire Plus during primary angioplasty for acute myocardial infarction. AB - OBJECTIVES: To evaluate the effectiveness of distal protection with the GuardWire Plus during primary angioplasty in patients with acute myocardial infarction. METHODS: Thirty-eight consecutive patients undergoing stent implantation with distal protection using the GuardWire Plus (DP-group) were compared with a matched control group undergoing conventional stent implantation after balloon angioplasty without distal protection (NDP-group). Microvascular circulation after revascularization was assessed by Thrombolysis in Myocardial Infarction (TIMI) flow grade, myocardial blush grade (MBG), serum creatine kinase peak release, and ST resolution. Left ventricular ejection fraction was measured by echocardiography at discharge. Follow-up quantitative coronary angiography and left ventriculography were performed 6 months after percutaneous coronary intervention. Quantitative coronary angiography data, restenosis rate, target lesion revascularization rate and follow-up left ventricular ejection fraction were also compared between the two groups. RESULTS: No significant differences were observed in baseline clinical and angiographic characteristics between the two groups. The TIMI flow grade 3 (DP-group 81.6% vs NDP-group 57.9%)and MBG 3 (57.9% vs 30.6%)were significantly greater in the DP-group respectively (p < 0.05). Post procedural ST-segment resolution > or = 50% was found in a significantly higher percentage of patients in the DP-group (68.4% vs 42.1%, p < 0.05). Left ventricular ejection fraction at discharge was significantly greater in the DP-group (55.5 +/- 8.5% vs 45.7 +/- 11.1%, p < 0.05). However, 6 months after the percutaneous coronary intervention, no significant difference was observed between the two groups. Restenosis rate and target lesion revascularization rate were similar in the two groups. CONCLUSIONS: Distal protection with the GuardWire Plus improved the microvascular circulation as assessed by TIMI flow grade, MBG, and ST resolution. Furthermore, left ventricular ejection fraction at discharge was improved. PMID- 15801275 TI - Combined treatment with statin and angiotensin-receptor blocker after stenting as a useful strategy for prevention of coronary restenosis. AB - OBJECTIVES: 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin) enhances the inhibitory effects of angiotensin-receptor blocker (ARB) on vascular neointimal formation in mice. The present case-control study investigated the efficacy of combined treatment with statin and ARB for preventing restenosis in patients with coronary artery disease. METHODS: We examined 210 patients with angina pectoris undergoing elective coronary stenting for de novo lesions of native coronary arteries. All enrolled patients received aspirin and ticlopidine. The subjects included patients who received no statin (control group, n = 137) or started statin treatment (statin group, n = 73) with or without ARB treatment after stenting. RESULTS: The rate of restenosis at 6 months after stent implantation in the statin group (19%) was significantly lower than that in the control group (32%). The restenosis rate in self-expanding Radius stents. (23%) was significantly lower than that in balloon-expandable Velocity stents (42%) independent of statin treatment. Patients treated with statins and ARBs were least likely [odds ratio (95% confidence interval): 0.30 (0.12-0.74)] to develop coronary restenosis, as assessed by multiple logistic regression analysis. CONCLUSIONS: These findings indicate that combined treatment with statin and ARB after stenting is a useful strategy for the prevention of coronary restenosis. PMID- 15801276 TI - [Coronary artery embolism from ruptured plaque in the left main trunks with difficulty in detection of culprit lesion: a case report]. AB - A 60-year-old man complained of severe chest pain and was emergently admitted to our hospital with a dignosis of anterior acute myocardial infarction. Emergent coronary angiography revealed significant stenosis in segment 7 and filling defect in segment 11 without flow delay. Haziness was observed in segment 5. Coronary thromboembolism was suspected, but the embolic source or culprit lesion was hard to detect. Intravascular ultrasonography detected ruptured plaque with lipid pooling in segment 5. Stent implantation for segment 5 was performed successfully and the patient had an excellent clinical course. Coronary thromboembolism is rare and intravascular ultrasonography may be useful to detect the culprit lesion. PMID- 15801277 TI - Three episodes of inferior acute myocardial infarction due to thrombotic occlusion of the right coronary artery: a case report. AB - A 77-year-old female with two previous inferior myocardial infarctions was transferred to our medical center with a third inferior acute myocardial infarction. Coronary angiography revealed 99% stenosis with rich thrombus in the distal right coronary artery [Thrombolysis in Myocardial Infarction (TIMI) grade 2 flow]. The angiographic appearance of the right coronary artery was similar to the two previous myocardial infarctions. Coronary aspiration was performed and TIMI grade 3 flow was established. To confirm the presence of thrombus, intravascular ultrasound (IVUS) and coronary angioscopy were performed at pre discharge. IVUS showed a thrombus-like low-density area at the mid right coronary artery. Red thrombi were observed in the same area using coronary angioscopy. Although warfarin had been prescribed for secondary prevention since the first acute myocardial infarction, both the second and third acute myocardial infarction occurred after cessation of warfarin. Patients with acute myocardial infarction due to thrombotic occlusion, confirmed by IVUS or angioscopy, might be good candidates for permanent warfarin therapy. PMID- 15801278 TI - Recurrent cardiogenic shock caused by dynamic mitral regurgitation in a patient with hypertrophic obstructive cardiomyopathy: a case report. AB - A 68-year-old man with hypertrophic obstructive cardiomyopathy developed recurrent cardiogenic shock due to dynamic mitral regurgitation. The pressure gradient in the left ventricular outflow tract under medication was 30 mmHg, and he complained of no symptom. He developed shock with dyspnea suddenly after bathing and defecation. Echocardiography and left ventriculography revealed massive mitral regurgitation and mild increment of pressure gradient in the left ventricular outflow tract (50 mmHg). He underwent successful mitral valve replacement following treatment with beta-blocker under intraaortic balloon pumping support. This case illustrates that exacerbation of the systolic anterior motion of the mitral anterior leaflet can cause dynamic severe mitral regurgitation with 'mild' increment of pressure gradient in the left ventricular outflow tract, resulting in cardiogenic shock with severe lung edema. PMID- 15801279 TI - [Diastolic abnormal flow in the left ventricular outflow tract observed in a 25 year-old woman]. PMID- 15801280 TI - [Guidelines for the diagnosis, treatment and prevention of pulmonary thromboembolism and deep vein thrombosis (JCS 2004)]. PMID- 15801281 TI - [Clinical evaluation of meningeal carcinomatosis associated with primary lung cancer]. AB - We evaluated diagnosis and treatment of four cases of meningeal carcinomatosis associated with primary lung cancer: case 1; small cell carcinoma (64 years old), case 2; small cell carcinoma (50 years old), case 3; adenocarcinoma (53 years old), and case 4; adenocarcinoma (55 years old). Determination of tumor markers in cerebrospinal fluid (CSF) together with the MRI findings that Gd-DTPA-enhanced T1-weighted image showing high intensity signal along the spinal cord was clinically useful in the diagnosis of meningeal carcinomatosis. Two of four patients received intrathecal chemotherapy and/or CSF drainage through Ommaya Reservoir, resulting in dramatic improvement of various symptoms such as motor weakness and vesicorectal disorder. Intrathecal chemotherapy and placement of an Ommaya-Reservoir for CSF drainage should be considered to provide better Quality of Life (QOL) when patient can tolerate it. PMID- 15801282 TI - [Factors affecting the success or failure of smoking cessation using nicotine patches]. AB - We identified the initial diagnostic factors that influenced the success or failure of patients trying to quit smoking using nicotine patches. In a smoking cessation treatment program at a smoking clinic, each patient received about 30 minutes of counseling in the initial diagnosis, then undertook a 2-month smoking cessation program using the nicotine patch. Between March 2000 and June 2002, 45 patients consulted the clinic. We attempted to monitor 30 patients whose smoking status we were able to observe. The patient group consisted of 5 women and 25 men who ranged in age from 22 to 75 years (mean age, 49 years). A follow-up survey by telephone was carried out (median follow-up time: 184.5 days). Actuarial smoking cessation curves were calculated according to the Kaplan-Meier method, and comparisons were made with the generalized Wilcoxon test. The Cox proportional hazards model was used for multivariate analysis. At the end of the two-month period, 86.3% of the patients had not resumed smoking; at one year after the program began, 56.7% had not resumed smoking. In the univariate analysis, the significant factors in the failure to maintain cessation were: a smoking start age of under 18 years, no affective disease, and smoking the day's first cigarette within 5 minutes after waking up (p < 0.05). In the multivariate analysis, the independent predictive factors in failure were: a starting smoking age of under 18 years and no affective disease (p < 0.05). Thus, patients who started smoking at a young age or who were free of affective disease were more likely to fail in their attempt to quit smoking. Attention to these factors is necessary as part of the guidance provided for smoking cessation. PMID- 15801283 TI - [Carbamazepine-induced pneumonitis definitively diagnosed by accidental readministration]. AB - An 82-year-old man had been treated by carbamazepine for convulsions. A month later he felt febrile and malaise. Laboratory data revealed liver dysfunction, hypoxemia, and chest radiograph and computed tomography (CT) of the thorax showed ground glass opacity in both lungs and mediastinal lymphadenopathy. Analysis of bronchoalveolar lavage fluid revealed pulmonary lymphocytosis. Drug-induced lymphocyte stimulation test (DLST) for carbamazepine using peripheral blood lymphocytes was negative. We gave him a possible diagnosis of carbamazepine induced liver damage and pneumonitis. After stopping carbamazepine, we initiated corticosteroid therapy and he recovered well. One year later he again suffered from convulsions and was treated with carbamazepine in another hospital. A week later he became febrile and suffered appetite loss, and came to our hospital. His chest radiograph and CT again showed ground glass opacity and pleural effusion in both lungs. Along with the laboratory data we established a definitive diagnosis of carbamazepine-induced pneumonitis. We discuss the mechanism of carbamazepine induced pneumonitis in which the DLST results were negative and the challenge test was positive. PMID- 15801284 TI - [A case of Kartagener's syndrome]. AB - This case describes a 57-year-old woman in whom situs inversus had been noted at her birth. She had bronchial asthma and bilateral sinusitis during her childhood. She married and experienced childbirth. In December 2003, she was admitted to our Division complaining of wheezing, expectoration and dyspnea on effort. Bronciectasis was visualized on chest X-ray and CT. Electron microscopic examination of the nasal cavity epithelium and bronchial epithelial cilia revealed a deficit of bilateral dynein arms. These findings, helped establish a diagnosis of Kartagener's syndrome, which is characterized by primary ciliary dyskinesia. The restrictive and obstructive pulmonary dysfunction with increase of residual volume in the lung function tests and diffuse centrilobular small nodules with hyperinflation on chest CT were consistent with the findings of diffuse panbronchilitis (DPB) and suggested extended obliterative peripheral airway disease. Clarithromycin which is highly effective for DPB failed to prevent the aggravation of airway infection, arousing the concern about the progression into chronic respiratory failure. PMID- 15801285 TI - [A case of successful medical treatment for necrotizing fasciitis of the chest wall with diabetic nephropathy]. AB - A 52-year-old man was given a diagnosis of type 2 diabetes mellitus at age 39. At age 46, he stopped taking medication. Two weeks after burning his legs at low temperature, he fell, using his right arm to protect his legs. The next day, he complained of pain and slight swelling from his right shoulder to his anterior chest and came to our hospital. At that time, a plain computed tomography scan suggested gasogenic bacterial infection and we discussed the indications for debridment. Although his widespread inflammation required extensive treatment including thoracostomy, we abandoned surgical treatment and administered several antibiotics in appropriate combination because of his severe condition. After admission, the mass grew rapidly and it was diagnosed as necrotizing fasciitis based on percutaneous needle biopsy and clinical findings. Although both inflammatory reactions and mass size tended to improve, he had repeated recurrence of pain and swelling in his right anterior chest. When he had a second recurrence, he received additional short-term steroid therapy. Afterwards he had no further recurrence. In this case, early clinical diagnosis, using broad spectrum antibiotics prior to definite diagnosis, and additional short-term steroid therapy at the time of the recurrence were effective. PMID- 15801286 TI - [A case of pulmonary parenchymal endometriosis]. AB - In 1982, a 45-year-old woman was hospitalized because of an abnormal lung shadow. The chest film provided evidence of pulmonary nodules. She was asymptomatic and had no labolatory abnormalities. Exploratory excision was performed to further assess the pulmonary nodules. The initial diagnosis was a suspected "hemangiopericytoma". She received combination chemotherapy but it was not effective. The nodules increased and decreased in size and number over the next decade from 1982, reaching a maximum by 1990 and a minimum by the end of 1992. However, her clinical condition remained stable during this period. Retrospective immunohistochemical analysis of CD10 and progesterone receptors confirmed a diagnosis of pulmonary parenchymal endometriosis. This was an unusual case of pulmonary parenchymal endometriosis that demonstrated various changes in the size and number of nodules without symptoms. There are few previous reports that demonstrated such nodular variation without symptoms over the course of disease. PMID- 15801287 TI - [A case of corticosteroid-resistant nonspecific interstitial pneumonia associated with dermatomyositis successfully treated with cyclosporin A]. AB - We report a case of corticosteroid-resistant nonspecific interstitial pneumonia associated with dermatomyositis. The patient was successfully treated with a combination of cyclosporin A (CsA) and prednisolone. A 28-year-old man was admitted complaining of skin eruptions, myalgia, and weakness, and dyspnea on exertion. Characteristic skin lesions such as erythema around the nails, teleangiectasis, and edema of the eyelids were observed. Dermatomyositis was diagnosed based on the diagnostic criteria, including the skin biopsy findings. Laboratory examination showed elevation of creatine kinase, but the anti-Jo-1 antibody was negative. Chest X-ray on admission showed reticular shadows and airspace consolidation mainly in both lower lung fields. Lung biopsy specimens taken under video-assisted thoracoscopic surgery revealed infiltration of lymphocytes into the thickened alveolar septa, and this revelation was compatible with the description of cellular nonspecific interstitial pneumonia. Administration of corticosteroids with steroid pulse therapy resulted in with improvement of interstitial pneumonia. However, after the combination therapy of corticosteroid with daily oral administration of 100-150 mg of CsA had been initiated, hypoxemia recovered swiftly in 5 days, and respiratory symptoms improved gradually, but steadily. Oral administration of corticosteroid and CsA was eventually tapered off and terminated completely 5 years later. The patient has been quite well with no treatment for two and a half years. The case presented here implies that CsA combined with corticosteroid may be efficacious in the treatment of interstitial pneumonia associated with dermatomyositis which is refractory to corticosteroid therapy. PMID- 15801288 TI - Declassifying coverage. New guidance documents issued by the CMS may clarify the great unknown of Medicare coverage determination. AB - Heard of Section 731 of the Medicare Modernization Act? The provision, meant to clear up the generally murky and unwieldy Medicare reimbursement approval process, could quicken the national approval process for medical procedures and devices. The changes will create a more definable approach to OK'ing new technologies. "I like the idea of bringing more science to bear," says Richard Pico, left. PMID- 15801289 TI - Personal and confidential. Lawmakers consider protecting identity of providers who report errors. PMID- 15801290 TI - JCAHO plans safety center. PMID- 15801291 TI - Taking it to the states. Bills would increase restrictions on niche facilities. PMID- 15801292 TI - Desperate states. Governors feel the heat to revamp Medicaid. PMID- 15801293 TI - Tough enough? Senate panel to decide if GPO plan has teeth. PMID- 15801294 TI - Budget showdown. Tug of war may ensue over planned Medicaid cuts. PMID- 15801295 TI - And another one. Healthcare exec nominated for seat on J&J board. PMID- 15801296 TI - IRS anxiety. Hospitals receiving executive compensation letters. PMID- 15801297 TI - Looking to boost Medicaid. Louisiana, Texas ponder taxing hospitals to aid besieged program. PMID- 15801298 TI - Corruption allegations renewed. Ill. influence-peddling scandal heats up with filing. PMID- 15801299 TI - CPOE is far from KO'd. JAMA study only proves that no technology cures problems in isolation. PMID- 15801300 TI - Mean what you say. Before achieving IT interoperability, leaders must agree on a definition. PMID- 15801301 TI - Trustees of the Year 2005. Leading the clean-up. PMID- 15801302 TI - Trustees of the Year 2005. Driving a system's success. PMID- 15801303 TI - Surgery centers warned of hospital lobby tactics. PMID- 15801304 TI - Robots: the next generation. Whether it's automation, transportation or advanced computerization, machines are hard at work in America's healthcare organizations. PMID- 15801305 TI - Coded excitation for diagnostic ultrasound: a system developer's perspective. AB - Resolution and penetration are primary criteria for clinical image quality. Conventionally, high bandwidth for resolution was achieved with a short pulse, which results in a tradeoff between resolution and penetration. Coded excitation extends the bounds of this tradeoff by increasing signal-to-noise ratio (SNR) through appropriate coding on transmit and decoding on receive. Although used for about 50 years in radar, coded excitation was successfully introduced into commercial ultrasound scanners only within the last 5 years. This delay is at least partly due to practical implementation issues particular to diagnostic ultrasound, which are the focus of this paper. After reviewing the basics of biphase and chirp coding, we present simulation results to quantify tradeoffs between penetration and resolution under frequency-dependent attenuation, dynamic focusing, and nonlinear propagation. Next we compare chirp and Golay code performance with respect to image quality and system requirements, then we show clinical images that illustrate the current applications of coded excitation in B mode, harmonic, and flow imaging. PMID- 15801306 TI - Coded excitation for synthetic aperture ultrasound imaging. AB - Peak acoustic power limits the signal-to-noise ratio (SNR) of real-time ultrasound images. For most conventional scan formats, however, the average power is well below heating limits. This means the SNR can be significantly increased using coded excitation. A coded system transmits a broadband, temporally elongated excitation pulse with a finite time-bandwidth product. The received signal must be decoded to produce an imaging pulse with improved SNR resulting from the higher average power in the elongated excitation. Decoding can produce significant range side lobes, however, greatly reducing image quality. All practical coding designs, therefore, represent a trade-off between SNR gain and range side lobes. A specific coding scheme appropriate for synthetic aperture imaging is presented. A 14.5 dB SNR improvement with acceptable range side lobes is demonstrated on a forward-looking imaging system appropriate for intravascular applications. PMID- 15801307 TI - Use of modulated excitation signals in medical ultrasound. Part I: Basic concepts and expected benefits. AB - This paper, the first from a series of three papers on the application of coded excitation signals in medical ultrasound, discusses the basic principles and ultrasound-related problems of pulse compression. The concepts of signal modulation and matched filtering are given, and a simple model of attenuation relates the matched filter response with the ambiguity function, known from radar. Based on this analysis and the properties of the ambiguity function, the selection of coded waveforms suitable for ultrasound imaging is discussed. It is shown that linear frequency modulation (FM) signals have the best and most robust features for ultrasound imaging. Other coded signals such as nonlinear FM and binary complementary Golay codes also have been considered and characterized in terms of signal-to-noise ratio (SNR) and sensitivity to frequency shifts. Using the simulation program Field II, it is found that in the case of linear FM signals, a SNR improvement of 12 to 18 dB can be expected for large imaging depths in attenuating media, without any depth-dependent filter compensation. In contrast, nonlinear FM modulation and binary codes are shown to give a SNR improvement of only 4 to 9 dB when processed with a matched filter. Other issues, such as depth-dependent matched filtering and use of filters other than the matched filter (inverse and Wiener filters) also are addressed. PMID- 15801308 TI - Use of modulated excitation signals in medical ultrasound. Part II: Design and performance for medical imaging applications. AB - In the first paper, the superiority of linear FM signals was shown in terms of signal-to-noise ratio and robustness to tissue attenuation. This second paper in the series of three papers on the application of coded excitation signals in medical ultrasound presents design methods of linear FM signals and mismatched filters, in order to meet the higher demands on resolution in ultrasound imaging. It is shown that for the small time-bandwidth (TB) products available in ultrasound, the rectangular spectrum approximation is not valid, which reduces the effectiveness of weighting. Additionally, the distant range sidelobes are associated with the ripples of the spectrum amplitude and, thus, cannot be removed by weighting. Ripple reduction is achieved through amplitude or phase predistortion of the transmitted signals. Mismatched filters are designed to efficiently use the available bandwidth and at the same time to be insensitive to the transducer's impulse response. With these techniques, temporal sidelobes are kept below 60 to 100 dB, image contrast is improved by reducing the energy within the sidelobe region, and axial resolution is preserved. The method is evaluated first for resolution performance and axial sidelobes through simulations with the program Field II. A coded excitation ultrasound imaging system based on a commercial scanner and a 4 MHz probe driven by coded sequences is presented and used for the clinical evaluation of the coded excitation/compression scheme. The clinical images show a significant improvement in penetration depth and contrast, while they preserve both axial and lateral resolution. At the maximum acquisition depth of 15 cm, there is an improvement of more than 10 dB in the signal-to-noise ratio of the images. The paper also presents acquired images, using complementary Golay codes, that show the deleterious effects of attenuation on binary codes when processed with a matched filter, also confirmed by presented simulated images. PMID- 15801309 TI - Use of modulated excitation signals in medical ultrasound. Part III: High frame rate imaging. AB - This paper, the last from a series of three papers on the application of coded excitation signals in medical ultrasound, investigates the possibility of increasing the frame rate in ultrasound imaging by using modulated excitation signals. Linear array-coded imaging and sparse synthetic transmit aperture imaging are considered, and the trade-offs between frame rate, image quality, and SNR are discussed. It is shown that FM codes can be used to increase the frame rate by a factor of two without a degradation in image quality and by a factor of 5, if a slight decrease in image quality can be accepted. The use of synthetic transmit aperture imaging is also considered, and it is here shown that Hadamard spatial encoding in transmit with FM emission signals can be used to increase the frame rate by 12 to 25 times with either a slight or no reduction in signal-to noise ratio and image quality. By using these techniques a complete ultrasound phased array image can be created using only two emissions. PMID- 15801310 TI - Spatio-temporal coding in complex media for optimum beamforming: the iterative time-reversal approach. AB - Spatio-temporal encoding in transmit and receive modes is of major importance in the development of ultrasound imaging devices. Classically, the assumption of constant sound speed in the medium allows one to restrict the beamforming process to the application of a cylindrical time-delay law on the elements of a multiple transducer array. Here is proposed an iterative time-reversal method capable of taking into account all the heterogeneities of the medium, concerning density, speed of sound, and absorption variations. It will be shown that this iterative focusing process converges toward a spatio-temporal inverse filter focusing, the first step of the process being a time-reversal focusing on the targeted point. This method can be seen as a calibration process and has been successfully applied to transskull focusing and intraplate echoes suppression. It is leading the way to promising applications such as high-resolution ultrasonic brain imaging and high-resolution focusing through complex reverberating media, in nondestructive testing and telecommunications. This work highlights the advantages of using spatio-temporal coding to focus through complex media. Such codes require the use of fully programmable, multichannel electronics to implement this technique in real time. PMID- 15801311 TI - Coded pulse excitation for ultrasonic strain imaging. AB - Decorrelation strain noise can be significantly reduced in low echo-signal-to noise (eSNR) conditions using coded excitation. Large time-bandwidth-product (>30) pulses are transmitted into tissue mimicking phantoms with 2.5-mm diameter inclusions that mimic the elastic properties of breast lesions. We observed a 5 10 dB improvement in eSNR that led to a doubling of the depth of focus for strain images with no reduction of spatial resolution. In high eSNR conditions, coded excitation permits the use of higher carrier frequencies and shorter correlation windows to improve the attainable spatial resolution for strain relative to that obtained with conventional short pulses. This paper summarizes comparative studies of strain imaging in noise-limited conditions obtained by short pulses and four common aperiodic codes (chirp, Barker, suboptimal, and Golay) as a function of attenuation, eSNR and applied strain. Imaging performance is quantified using SNR for displacement (SNRd), local modulation transfer function (LMTF), and contrast-to-noise ratio for strain (CNRepsilon). We found that chirp and Golay codes are the most robust for imaging soft tissue deformation using matched filter decoding. Their superior performance is obtained by balancing the need for low-range lobes, large eSNR improvement, and short-code duration. PMID- 15801312 TI - Harmonic chirp imaging method for ultrasound contrast agent. AB - Coded excitation is currently used in medical ultrasound to increase signal-to noise ratio (SNR) and penetration depth. We propose a chirp excitation method for contrast agents using the second harmonic component of the response. This method is based on a compression filter that selectively compresses and extracts the second harmonic component from the received echo signal. Simulations have shown a clear increase in response for chirp excitation over pulse excitation with the same peak amplitude. This was confirmed by two-dimensional (2-D) optical observations of bubble response with a fast framing camera. To evaluate the harmonic compression method, we applied it to simulated bubble echoes, to measured propagation harmonics, and to B-mode scans of a flow phantom and compared it to regular pulse excitation imaging. An increase of approximately 10 dB in SNR was found for chirp excitation. The compression method was found to perform well in terms of resolution. Axial resolution was in all cases within 10% of the axial resolution from pulse excitation. Range side-lobe levels were 30 dB below the main lobe for the simulated bubble echoes and measured propagation harmonics. However, side-lobes were visible in the B-mode contrast images. PMID- 15801313 TI - Design and preliminary tests of a family of adaptive waveforms to measure blood vessel diameter and wall thickness. AB - In this article we consider the adaptive design of waveforms to be used in vascular ultrasound. The advantage of these waveforms, when used with the proposed processing scheme, is that their application results in increased reflected energy, especially when compared with more conventional methods such as a short-gated sinusoid. This increase in reflected energy has potential to permit inferences to be made about wall thickness and vessel diameter from deeper vessels than possible with more traditional techniques. Here, the use of waveforms of the type A(t)ej(kt2), 0 < or = t < or = b, where A(t) is a specially designed envelope and k a sweep frequency, is proposed. Theorems are proved that describe how to choose an A(t) which results in either a maximum of reflected energy signal-to-noise ratio (SNR), or range resolution. The design of the waveform is adaptive in that both A(t) and k are derived in consideration of a specific blood vessel whose transfer function has been obtained experimentally. Numerical simulations illustrate the advantages of using these waveforms as well as the effects of the parameters. A simple experimental implementation of the methodology is presented on a brachial artery. The measurement of the impulse response of the artery is presented in this context. Results indicate that a processing gain in SNR over the instantaneous values obtained from the raw echo waveforms of 11 dB to 14 dB can be obtained via this methodology. PMID- 15801314 TI - ICARUS: imaging pulse compression algorithm through remapping of ultrasound. AB - In this work we tackle the problem of applying to echographic imaging those synthetic aperture focusing techniques (SAFT) in the frequency domain commonly used in the field of synthetic aperture radars (SAR). The aim of this research is to improve echographic image resolution by using chirp transmit signals, and by performing pulse compression in both dimensions (depth and lateral). The curved geometry present in the unfocused radio-frequency (RF) ultrasonic image is the main cause of inaccuracy in the direct application of frequency domain SAFT algorithms to echographic imaging. The focusing method proposed in this work, after pulse compression in the depth dimension, performs lateral focusing in the mixed depth-lateral spatial frequency domain by means of a depth variant remapping followed by lateral pulse compression. This technique has the advantage of providing a resolution that is uniform in nonfrequency selective attenuation media, and improved with respect to conventional time domain SAFT, without requiring the acquisition and processing of channel data necessary for the most advanced synthetic transmit aperture techniques. Therefore, the presented method is suitable for easy real-time implementation with current generation hardware. PMID- 15801315 TI - Noncontact, high-resolution ultrasonic imaging of wood samples using coded chirp waveforms. AB - A noncontact ultrasonic inspection technique has been developed to study the properties of wood samples in air. The system makes use of two broad bandwidth capacitive transducers, combined with signal processing techniques. A coded chirp signal was used in the current application to provide a waveform that could be postprocessed to provide sufficient sensitivity for transmission across samples of wood. It will be shown in this paper that the signal-to-noise ratio (SNR) can be greatly improved using two signal recovery techniques, namely pulse compression and swept frequency multiplication (SFM). A simulation of both techniques is presented and compared to experimental data. As will be seen from the experimental results, it is possible to perform noncontact ultrasonic experiments to extract a range of useful information such as ring density and the presence of microcracks. PMID- 15801316 TI - The clock model and its relationship with the Allan and related variances. AB - The clock errors are modeled by stochastic differential equations (SDE) and the relationships between the diffusion coefficients used in SDE and the Allan variance, a typical tool used to estimate clock noise, are derived. This relationship is fundamental when a mathematical clock model is used, for example in Kalman filter, noise estimation, and clock prediction activities. PMID- 15801317 TI - Coupled determination of gravimetric and elastic effects on two resonant chemical sensors: love wave and microcantilever platforms. AB - The objective of this paper is to couple theoretical and experimental results from microcantilevers and Love-wave acoustic devices in order to identify and separate mass loading effects from elastic effects. This is important in the perspective of sensing applications. For that, a thin-film polymer is deposited on both resonant platforms. It is demonstrated that microcantilevers are essentially mass sensitive. They allow one to determine the polymer layer thickness, which is validated by optical profilometry measurements. Then, taking into account this thickness, theoretical modeling and experimental measurements with Love-wave devices permit one to estimate an equivalent elastic shear modulus of the thin-film polymer at high frequency. Results are interesting if one is to fully understand and optimize (bio)chemical sensor responses. PMID- 15801318 TI - Response of fiber Bragg gratings to longitudinal ultrasonic waves. AB - In the last years, fiber optic sensors have been widely exploited for several sensing applications, including static and dynamic strain measurements up to acoustic detection. Among these, fiber Bragg grating sensors have been indicated as the ideal candidate for practical structural health monitoring in light of their unique advantages over conventional sensing devices. Although this class of sensors has been successfully tested for static and low-frequency measurements, the identification of sensor performances for high-frequency detection, including acoustic emission and ultrasonic investigations, is required. To this aim, the analysis of feasibilty on the use of fiber Bragg grating sensors as ultrasonic detectors has been carried out. In particular, the response of fiber Bragg gratings subjected to the longitudinal ultrasonic (US) field has been theoretically and numerically investigated. Ultrasonic field interaction has been modeled, taking into account the direct deformation of the grating pitch combined with changes in local refractive index due to the elasto-optic effect. Numerical results, obtained for both uniform and Gaussian-apodized fiber Bragg gratings, show that the grating spectrum is strongly influenced by the US field in terms of shape and central wavelength. In particular, a key parameter affecting the grating response is the ratio between the US wavelength and the grating length. Normal operation characterized by changes in wavelength of undistorted Bragg peak is possible only for US wavelengths longer than the grating length. For US wavelengths approaching the grating length, the wavelength change is accompanied by subpeaks formation and main peak amplitude modulation. This effect can be attributed to the nonuniformity of the US perturbation along the grating length. At very high US frequencies, the grating is not sensitive any longer. The results of this analysis provide useful tools for the design of grating-based ultrasound sensors for meeting specific requirements in terms of field intensity and frequencies. PMID- 15801319 TI - Ultrasonic data compression via parameter estimation. AB - Ultrasonic imaging in medical and industrial applications often requires a large amount of data collection. Consequently, it is desirable to use data compression techniques to reduce data and to facilitate the analysis and remote access of ultrasonic information. The precise data representation is paramount to the accurate analysis of the shape, size, and orientation of ultrasonic reflectors, as well as to the determination of the properties of the propagation path. In this study, a successive parameter estimation algorithm based on a modified version of the continuous wavelet transform (CWT) to compress and denoise ultrasonic signals is presented. It has been shown analytically that the CWT (i.e., time x frequency representation) yields an exact solution for the time-of arrival and a biased solution for the center frequency. Consequently, a modified CWT (MCWT) based on the Gabor-Helstrom transform is introduced as a means to exactly estimate both time-of-arrival and center frequency of ultrasonic echoes. Furthermore, the MCWT also has been used to generate a phase x bandwidth representation of the ultrasonic echo. This representation allows the exact estimation of the phase and the bandwidth. The performance of this algorithm for data compression and signal analysis is studied using simulated and experimental ultrasonic signals. The successive parameter estimation algorithm achieves a data compression ratio of (1-5N/J), where J is the number of samples and N is the number of echoes in the signal. For a signal with 10 echoes and 2048 samples, a compression ratio of 96% is achieved with a signal-to-noise ratio (SNR) improvement above 20 dB. Furthermore, this algorithm performs robustly, yields accurate echo estimation, and results in SNR enhancements ranging from 10 to 60 dB for composite signals having SNR as low as -10 dB. PMID- 15801320 TI - Capacitive micromachined ultrasonic transducer design for high power transmission. AB - Capacitive micromachined ultrasonic transducers (cMUTs) were developed to meet the demands of the ultrasonic industry. To achieve maximum efficiency, the conventional operation of the cMUT requires a bias voltage close to the collapse voltage. Total acoustic output pressure is limited by the efficiency of the cMUT and the maximum-allowed pulse voltage on the membrane. In this paper, we propose the collapse-snapback operation of the cMUT: the membrane is collapsed onto the substrate in the collapsing cycle, and released in the snapback cycle. The collapse-snapback operation overcomes the above-mentioned limitations of the conventional operation. The collapse-snapback operation utilizes a larger range of membrane deflection profiles (both collapsed and released profiles) and generates higher acoustic output pressures. The static finite element calculations were performed to design cMUTs with specific collapse and snapback voltages by changing the electrode parameters (radius (re), position (de), and thickness (te)). These designs were refined for optimum average displacement per cycle. An electrode radius greater than 60% of the membrane radius significantly improved the displacement per volt. Moderately thick membranes (te approximately 0.2 microm) were preferred, as thicker membranes reduced the displacement per volt. Under proper bias conditions, the collapse-snapback operation, designed for high-power transmission, allowed the application of pulse voltages larger than the difference of collapse and snapback voltages. Dynamic finite element calculations of an infinite cMUT array on the substrate loaded with acoustic fluid medium were performed to determine the dynamic response of the cMUT. Commercially available FEM packages ANSYS and LS-DYNA were used for static and dynamic calculations, respectively. The cMUTs were fabricated for optimal performance in the collapse-snapback operation. The transmit experiments were performed on a 2-D cMUT array using a calibrated hydrophone. Taking into account the attenuation and diffraction losses, the pressure on the cMUT surface was extracted. The cMUT generated 0.47 MPa (6 kPa/V) and 1.04 MPa (11 kPa/V) in the conventional and collapse-snapback operations, respectively. Therefore, collapse snapback operation of the cMUTs was superior for high-power transmission. PMID- 15801321 TI - Current thinking on the relationship between rhinosinusitis and asthma. AB - An observation made initially on clinical grounds and epidemiologic evidence, i.e., that rhinosinusitis and asthma are closely linked diseases is now supported by a growing body of scientific evidence. Most recent evidence supports the characterization of rhinosinusitis and asthma as two compartmental expressions of a common mucosal susceptibility to exogenous stimuli. In addition, there is evidence that the compartmental processes can affect and amplify each other via a systemic intermediary. The bone marrow is involved in this process, and IL-5 may be a key cytokine for orchestrating the systemic interaction. These facts argue that rhinosinusitis and asthma are not simply localized disease processes but part of a systemic inflammatory disease affecting the respiratory tract. They also provide a compelling rationale for combined treatment strategies with consideration of the treatment of rhinosinusitis as a means of improving asthma control and monitoring for signs of bronchial involvement in those with rhinosinusitis. PMID- 15801322 TI - Prevalence of asthma and asthma-like symptoms in inner-city schoolchildren. AB - This study investigates the prevalence of asthma and asthma-related symptoms in New Orleans inner-city schoolchildren. A cross-sectional survey of 1535 elementary, middle, and high school children (aged 5-18) was conducted by using the International Study of Asthma and Allergies in Childhood (ISAAC) written questionnaire. Lifetime prevalence of wheezing was 39.4%, and lifetime prevalence of asthma was 24.4%. Wheezing during the previous 12 months was reported by 25.7% of the sample. Twenty-one percent of respondents reported having one or more attacks of wheezing per year, with 5.6% reporting four or more attacks per year. Many participants reported sleep disturbance (15.4%), with 6.2% reporting sleep disturbance more than once a week. The 12-month rate of speech limitation due to asthma exacerbation was 6.6%. Exercise-induced asthma was reported by 16.9% of the students, and nocturnal cough (not associated with cold) was reported by 27.3%. Overall, boys reported higher rates of symptoms than girls, and younger children (aged 6-7) reported greater symptoms than older children (aged 13-14). These findings show that prevalence of asthma in this population is elevated, and the ISAAC written questionnaire successfully identified inner-city children at risk for asthma in New Orleans. PMID- 15801323 TI - Peak expiratory flow variability and exercise responsiveness in methacholine hyperresponsive adolescents with asthma remission. AB - The aim of this study was to investigate whether bronchial hyperresponsiveness in adolescents with long-term asthma remission is associated with increased peak expiratory flow (PEF) variability and/or increased bronchial response to exercise (BRE). Twenty-nine adolescents with asthma remission (neither symptoms nor any medication used during the previous two years), but with persistent methacholine hyperresponsiveness (PC20 < 18 mg/mL; remission group), 29 methacholine PC20 matched adolescents with symptomatic asthma (symptomatic group), and 20 healthy subjects (control group) were studied. Subjects recorded PEF twice daily for 14 days and PEF variability, expressed as amplitude % mean, was calculated. Subjects also underwent a standardized exercise challenge; BRE was defined as a maximal % fall in FEV1 within 30 min after exercise. The mean (+/- SD) PEF variations in the symptomatic group and in the remission group were 12.10 +/- 6.35% and 10.02 +/- 4.73%, respectively, which were significantly higher than that (5.94 +/- 2.44%) of the control group. On the other hand, the degree of BRE (7.36 +/- 3.85%) in the remission group was significantly lower than that (22.31 +/- 10.50%) of the symptomatic group, and similar to that (5.98 +/- 2.70%) of the control group. Methacholine hyperresponsiveness in asthma remission during adolescence is associated with increased PEF variability but not with increased BRE. PMID- 15801324 TI - Atopic and nonatopic asthma in children. AB - In 155 asthmatic children we have studied the relationship between prick test positivity and a set of genetic factors previously found to be associated with bronchial asthma. Among these factors, MN system (p = 0.009) and age at onset of symptoms (p = 0.05) are the most important variables separating prick test negative from prick test positive children. MN and age at onset influence independently prick test positivity pointing to an additive effect of the two variables. M phenotype appears correlated positively with an increased susceptibility to nonallergic asthma in all age groups, whereas N phenotype appears correlated positively with age at onset but in allergic asthma only. The MN system codifies for glycophorin A, a sialoglycoprotein that represents a major ligand for several bacteria and viruses that recognize the N-acetylneuraminic acid present in this protein. The present data suggest that genetic variability in this system might influence bacterial and viral competition and mucosal damage influencing susceptibility to asthmatic reactions in absence of IgE hyperproduction. PMID- 15801325 TI - A respiratory therapist-directed protocol for managing inpatients with asthma and COPD incorporating a long-acting bronchodilator. AB - This prospective study was designed to determine whether incorporating formoterol into a standardized respiratory therapist-directed protocol for administering bronchodilators to hospitalized patients with obstructive airway disease would reduce health care resource use and provide a safety advantage. All patients admitted to Washington Hospital Center with asthma and chronic obstructive pulmonary disease (CODP) are administered bronchodilators under a standardized respiratory therapist-directed protocol. Formoterol was the primary bronchodilator for the intervention period from January through March 2002, with levalbuterol, albuterol, and ipratropium available as needed. Results for the intervention period were compared against two historical control periods. From January through March 2000, the bronchodilators in the protocol were albuterol and ipratropium, and from January through March 2001 levalbuterol, albuterol, and ipratropium were available. Health care resource use was determined by the number of bronchodilator treatments administered per admission. Costs (adjusted to 2002 dollars) for supplies, therapist time, and drugs were calculated for the three time periods. Adverse events related to bronchodilator administration were recorded in a standardized manner for all three time periods. Bronchodilator treatments per admission, respiratory therapist visits per admission, and time spent per admission, and cost per bronchodilator treatment significantly decreased in 2002. Significantly fewer adverse events related to bronchodilator treatments were reported in 2002 than 2000. The addition of formoterol to a respiratory therapist-directed protocol for administering bronchodilators reduced health care resource use and adverse events for patients with asthma and COPD. PMID- 15801326 TI - Outcomes associated with initiation of different controller therapies in a Medicaid asthmatic population: a retrospective data analysis. AB - BACKGROUND: Outcomes in asthmatic patients may vary depending on the controller medication used. Observational studies of outcomes of asthma therapy are needed to understand the implications of choice of controller in different populations. OBJECTIVES: To determine whether there are differences in health care use and costs of asthma treatment in asthma patients treated with montelukast compared with fluticasone proponiate 44 microg. METHOD: Using data from the North Carolina Medicaid program, we compared continuously enrolled asthmatic patients starting either fluticasone propionate 44 microg (FP44), an inhaled corticosteroid (ICS) (n = 312), or montelukast 5 and 10 mg, an oral leukotriene modifier (LM) (n = 398) between the years 1998 and 1999. A secondary analysis compared continuously enrolled asthmatic patients already using ICS as controller therapy initiating either salmeterol (long-acting beta-agonist) (n = 97) or montelukast (n = 101) in the year 1998. Patients were followed for 1 year pre- and postcontroller or additional controller initiation for health care service use, medication refill patterns, and costs. RESULTS: There were no significant differences in the adjusted asthma-related health care costs between the montelukast and FP44 groups. In both groups, physician visits were significantly higher in year 2 (p < 0.01) than in year 1. We found montelukast users to be more adherent with prescription refills (using measures of medication possession) even after allowing for a wider adherence range for FP (RR = 2.53; 95% CI = 1.50-4.26), although patients using montelukast were more likely than patients with fluticasone to switch controller pharmacotherapy (RR = 1.53; 95% CI = 1.12-2.09). Similarly, there were no differences in health care service use and costs between the montelukast and salmeterol groups, with the exception of a 33% reduction (p < 0.01) in number of inhaled corticosteroid refills in the second year in the salmeterol group. CONCLUSION: There were no cost and major health care use differences between the two primary or secondary controller therapies in the postinitiation year. Although FP was associated with lower rate of controller switch, montelukast use was associated with significantly better treatment adherence in patients with treatment persistence in this population of Medicaid enrolled asthmatic patients. The addition of salmeterol as additional controller was associated with a significant decrease in inhaled corticosteroid use, suggesting decreased adherence in patients on the two-drug regimen. PMID- 15801327 TI - Validation of a simplified quality-of-life questionnaire for socioeconomically deprived asthma patients. AB - INTRODUCTION: Quality-of-life questionnaires have been recognized as an important tool to measure the impact of asthma in the patient's life and has become a main outcome in clinical research. To be effective, questionnaires should be adapted to reflect the needs of the target population. OBJECTIVE: To assess the reliability, responsiveness, and cross-sectional validity of a simplified quality of-life questionnaire (QQL-EPM) as a tool specially developed for a socioeconomically deprived target population of asthmatic patients. METHOD: Thirty-five asthma patients were followed in a prospective open study over a period of 9 weeks. Clinical visits were performed monthly with pulmonary function assessment and the patients filled out a diary card regarding symptom scores, use of rescue medication, and PEF. At each visit, FEV1 and FVC were measured and two health-related quality-of-life questionnaires were applied: general quality of life (SF-36) and specific quality of life (QQL-EPM). The condition of patient regarding asthma control was assessed at each visit to the clinic, with treatment optimization and medication adjustment as needed. At the end of the study, each period was analyzed across the trial period and classified as stable or noncontrolled asthma. RESULTS: QQL-EPM was able to correlate changes in quality of life in patients with alterations in their asthma control condition (global = 0.0001) and to differentiate these patients from those whose condition remained stable (global = 0.0001). The reliability of QQL-EPM was 0.68-0.90, and correlation with other clinical measurements and generic quality of life was moderate. PMID- 15801328 TI - Increased oxidative stress in acute exacerbations of asthma. AB - Oxidant-antioxidant imbalance may play an important role in the pathogenesis of asthma, especially during acute exacerbations. To compare the systemic oxidant antioxidant status in patients with acute exacerbations and stable asthma, we measured a wide range of parameters of oxidant-antioxidant balance in leukocytes, plasma, and red cells of 32 patients with acute exacerbations and 97 patients with stable asthma. These included measurement of superoxide anion generation by leukocytes, lipid peroxidation (measured as TBARS), total nitrates and nitrites, protein carbonyls, and protein sulfhydryls in plasma. Antioxidant status was evaluated by measuring the red cell superoxide dismutase and catalase activity, total blood glutathione, glutathione peroxidase activity in red cell and plasma, and total antioxidant capacity (assessed as ferric reducing antioxidant power) in plasma. Plasma total antioxidant capacity and total protein sulfhydryls were found to be decreased (p < 0.01), whereas plasma lipid peroxides were found to be increased (p < 0.05), in acute exacerbations of asthma. No significant differences were found in plasma glutathione peroxidase, protein carbonyls, and total nitrates and nitrites, red cell antioxidative enzyme activities, superoxide anion release from leukocytes, and total blood glutathione between the two groups (p > 0.05). Our observations suggest that acute exacerbations of asthma are associated with increased oxidative stress that is evident from some of the parameters in the plasma. Failure to observe simultaneous changes in all parameters of oxidative stress may be due to the possibility of their responses being dissociated in time or compensatory changes occurring in some of these. PMID- 15801329 TI - Evaluation of asthma management policies in New York City public schools. AB - The aim of this study was to investigate asthma management policies and protocols in Bronx, New York, public elementary schools in light of the National Heart, Lung, and Blood Institute (NHLBI) Resolution on Asthma Management in Schools. An anonymous survey was conducted in November 2001 of principals, teachers, counselors, and nurses at five Bronx elementary schools. The response rate was 62%, and the majority of respondents (84%) were teachers; 51% of respondents learned of a student's asthma only through informal conversation with the student or parent, 28% said they were usually not informed of a student's asthma status, and only 10% learned of a child's asthma through existing school protocols; 21% of respondents did not know whom in the school was responsible for supervising the health needs of children with asthma, and 30% did not know how asthma inhalers were supposed to be handled at their school. Only 1.6% of teachers were "very familiar" with Board of Education asthma policies. The results are interpreted as showing poor adherence to the NHLBI recommendations. They also reflect a lack of consistent strategies for communication of a child's asthma diagnosis and for management of the disease in the schools. Ongoing efforts to improve asthma management in public schools through teacher education and policy development should be supported and evaluated for outcomes in teacher knowledge and student health. PMID- 15801330 TI - Inpatient management of asthma in the Chicago suburbs: the Suburban Asthma Management Initiative (SAMI). AB - BACKGROUND: Assessment of inpatient asthma management has generally been limited to urban settings, including Chicago, which is known for its high asthma morbidity and mortality. Previously published data have been based on survey methodology. The Suburban Asthma Consortium (SAC) sought to obtain patient-based data unique to the Chicago suburbs to improve asthma care in those areas. OBJECTIVE: To evaluate current inpatient asthma management based on the 1997 National Asthma Education and Prevention Program (NAEPP). DESIGN: Retrospective chart review of all hospitalized patients 3-65 years bearing asthma-related ICD-9 codes for fiscal year 2002 in community, nonteaching hospitals in Chicago suburbs. RESULTS: Nine hundred two cases were submitted from seven hospitals. The majority ( > or = 75%) received inhaled bronchodilators, systemic steroids, oxygen and pulse oximetry. Antibiotic use (67%), chest radiography (85%), complete blood count (77%), and electrolytes (59%) appeared excessive in view of NAEPP recommendations. Peak flow monitoring (PFM) was recorded on admission in 45% of patients 5 years old and older; 52% had PFM during hospitalization. Thirty eight percent of patients were taking ICS prior to admission; of those not on ICS, only 12% were newly diagnosed asthmatics. Overall, 51% of patients were discharged with ICS. Patients were more likely to receive ICS at discharge if they had required intensive care (ICU), had been on ICS prior to admission, were referred to an asthma specialist while hospitalized, or were insured. Patients with Medicare/Medicaid (MC/MA) had more repeat emergency visits and hospitalizations, longer lengths of stay, and received less ICS at discharge. Depending on the parameter, 41% or less patients received discharge planning education and were not more likely to have received education if in the ICU. Results ranged significantly between hospitals for most parameters (p < 0.05 or less). CONCLUSION: Study subjects received appropriate acute therapy and oxygen monitoring, but there was a divergence from NAEPP recommendations regarding PFM, ICS use, antibiotics, and laboratory evaluation. Patients receiving MC/MA experienced higher morbidity and received less ICS. Discharge asthma education was suboptimal for most hospitals. Most parameters demonstrated significantly wide practice variations between hospitals. Peak flow monitoring and patient education findings differed significantly from those in survey-conducted studies. PMID- 15801331 TI - Impact of a soluble phospholipase A2 inhibitor on inhaled allergen challenge in subjects with asthma. AB - The possible roles of secretory phospholipases A2 (sPLA2) in asthma include the release of arachidonic acid from cellular membranes, generation of lysophospholipids, sPLA2-mediated activation of cPLA2 with increased leukotriene production, and surfactant degradation. LY333013 is a potent inhibitor of sPLA2. This study examined the impact of two doses of LY333013 vs. placebo on allergen induced bronchoconstriction following inhaled allergen challenge in atopic asthmatics. Fifty subjects were randomly assigned to treatment, and 40 subjects completed the study. A double-blind, placebo-controlled, random order, crossover study design was used. LY333013 had no impact on the primary outcome variables of the areas under the FEV1 response curve early (0-3 hours) (AUC(early)) and late (3-8 hours) (AUC(Iate)) following inhaled allergen challenge. No significant drug related adverse effects were observed. The response to inhaled allergen challenge was reproducible and confirms the utility of this technique as a model in which to screen compounds for further testing in asthmatic patients. PMID- 15801332 TI - Soluble CD23 and interleukin-1 receptor antagonist in human asthmatics following antigen challenge. AB - Two postulated intrinsic anti-inflammatory mechanisms in asthma include the low affinity IgE receptor, or CD23, and interleukin 1 receptor antagonist (IL-1ra). We investigated the role these mediators play in the asthmatic response by measuring local levels in human asthmatics before and after segmental allergen challenge and examined the effect of inhaled corticosteroids on soluble CD23 and IL-1ra levels. Ten subjects underwent bronchoscopy at baseline and 24 hours after antigen challenge. Prior to challenge and every 12 hours afterward subjects received beclomethasone 252 microg or placebo. Fluid was analyzed for sCD23 and IL-1ra using ELISA immunoassays. Eosinophil percentages significantly increased at 24 hours following antigen challenge. sCD23 levels were generally undetectable at baseline and increased significantly following antigen challenge. IL-1ra levels increased 28-fold in the late-phase response. Beclomethasone significantly reduced the late-phase eosinophil percentage at 24 hours compared with placebo but did not attenuate late-phase sCD23 or IL-1ra levels. Our data showed a significant rise in the levels of two mediators thought to play an important role in the attenuation of the asthmatic response. The finding that steroid treatment did not enhance these levels suggests that this may be an independent approach to asthma therapy that should be investigated. PMID- 15801333 TI - The acute effect of methylphenidate in Brazilian male children and adolescents with ADHD: a randomized clinical trial. AB - OBJECTIVE: To evaluate the acute efficacy of methylphenidate (MPH) in Brazilian male children and adolescents with ADHD. METHOD: In a 4-day, double-blind, placebo-controlled, randomized, fix dose escalating, parallel-group trial, 36 ADHD children and adolescents were allocated to two groups: MPH (n = 19) and placebo (n = 17). Participants were evaluated pre- and posttreatment using the 10 item Conners Abbreviated Rating Scale (ABRS), the Children's Global Assessment Scale (CGAS), and a simplified version of the Continuous Performance Test (CPT). RESULTS: The MPH group had a significantly greater decrease in ABRS scores and a significantly higher increase in CGAS scores than the placebo group (p < 0.01). The MPH group showed also a significantly higher proportion of patients with a robust improvement (decrement of at least 50% in the ABRS score after the intervention) than the placebo group (p < 0.01). The MPH effect size for the ABRS was 1.05 (95% CI = 0.73-1.37). CONCLUSION: Our results extend the efficacy of MPH on the ADHD core symptoms extensively demonstrated in clinical trials with samples from some developed countries to a sample from a developing country where a diverse culture may modulate the clinical presentation of the disorder. PMID- 15801334 TI - Efficacy and safety of atomoxetine in childhood attention-deficit/hyperactivity disorder with comorbid oppositional defiant disorder. AB - OBJECTIVE: To compare the safety and efficacy of atomoxetine, a selective inhibitor of the norepinephrine transporter, versus placebo in Attention Deficit/Hyperactivity Disorder (ADHD) patients with comorbid Oppositional Defiant Disorder (ODD). METHODS: A subset analysis of 98 children from two identical, multi-site, double-blind, randomized, placebo-controlled trials involving 9 weeks of treatment with atomoxetine or placebo was conducted. Patients met DSM-IV ADHD criteria. ODD was diagnosed with the Diagnostic Interview for Children and Adolescents-IV (DICA-IV; Reich, Weiner, and Herjanic, 1997). ADHD severity was assessed with the ADHD Rating Scale-IV-Parent Version: Investigator Administered and Scored (ADHD-RS-IV-Parent:lnv; DuPaul, Power, Anastopoulos, and Reid, 1998); the short version of the Conners' Parent Rating Scales-Revised (CPRS-R:S; Conners, 2000); and the Clinical Global Impressions of ADHD Severity (CGI-ADHD-S; Guy, 1976). Clinical response was defined as a > or = 25% reduction in ADHD-RS-IV Parent:lnv total score. RESULTS: ADHD-RS-IV-Parent:lnv, CGI-ADHD-S, and three CPRS-R:S subscale scores improved markedly with atomoxetine treatment. However, a decrease in the CPRS-R:S Oppositional subscore for atomoxetine-treated patients was not significantly greater than scores for placebo-treated patients. Clinical response rates were 65.4% in the atomoxetine group, and 36.4% in the placebo group (p = .007). CONCLUSION: Atomoxetine was effective for the treatment of ADHD in patients with comorbid ODD. It did not significantly reduce the severity of ODD symptoms, and was well tolerated by the patients. PMID- 15801335 TI - Emotional reactivity in children with attention-deficit/hyperactivity disorder. AB - OBJECTIVE: This study examined Barkley's (1997b) theory regarding the emotional regulation of children with Attention-Deficit/Hyperactivity Disorder (ADHD). METHOD: Mothers of children with and without ADHD between the ages of 6 and 15 were asked to rate their child's emotional response on each of three measures. RESULTS: Children with ADHD were rated as significantly more emotionally reactive to both immediate and future events than were children without ADHD. Differences at both the immediate and future time periods were stronger in response to negative as opposed to positive emotional events. In response to the consequences of their behavior, however, children with ADHD were rated as less emotionally reactive than children without ADHD. DISCUSSION: Conclusions are made in reference to Barkley's theory and implications are explored. PMID- 15801336 TI - Diagnostic performance of the CBCL-Attention Problem Scale as a screening measure in a sample of Brazilian children with ADHD. AB - OBJECTIVE: To evaluate the diagnostic performance of the Attention Problem Scale of the Child Behavior Checklist (CBCL-APS) for the screening of Attention Deficit/Hyperactivity Disorder (ADHD) in a sample of Brazilian children and adolescents. METHODS: The CBCL-APS was given to 763 children and adolescents. Child psychiatrists using DSM-IV criteria confirmed the clinical diagnoses. Diagnostic performance was evaluated through Receiver-Operating Characteristic (ROC) curves. RESULTS: Only moderate areas under the curve (AUC) were found for the general sample (AUC = 0.79; 95% CI = 0.76-0.82), and for the subsample of referred patients (AUC = 0.78; 95% CI = 0.74-0.82). The subsample of patients with ADHD of the combined type presented the largest AUC (AUC = 0.85; 95% CI = 0.82-0.88). CONCLUSION: Our findings concur with previous studies of different cultures demonstrating adequate diagnostic performance of the CBCL-APS for the screening of ADHD, especially of the combined type. PMID- 15801337 TI - Promising indeed: the role of "experts" and practitioners in the introduction and use of new materials and techniques in restorative dentistry. AB - Contemporary general practitioners are facing enormous challenges. Whether they want to or not, they are forced to manage a complex small business and are often ill equipped educationally and emo- tionally to do so. They also have to manage a substantial number of staff members, often with complex emotional interactions. Young dentists today are frequently forced to service considerable debt as a result of educational costs and high practice overheads. Manufacturers and dental laboratories are aggressively marketing new products and procedures at an unprecedented rate, often with sophisticated marketing techniques. These practitioners also have lives outside of work as wives or husbands, fathers or mothers, scout leaders, and coaches. Although contemporary clinicians are faced with many challenges, they also must accept some responsibilities regarding new products and practices. They must possess critical thinking skills and a basic knowledge of materials science. Whether contemporary dental schools provide their students with an education that stimulates this is a topic for another editorial. Educationally prepared or not, practitioners should filter information through common sense and past experience. New products should be introduced to the practice carefully and used in a conservative manner. These overstressed individuals deserve the best possible information from those of us who are considered experts. We cannot continue to be politically correct when describing new products and materials. We need to call it the way it is. Those who are considered experts have achieved that status through a combination of hard work, talent, luck, and perhaps some sleight of hand. There are legitimate rewards to be reaped as an expert, but with those come responsibilities. Referring to products with no clinical testing or evidence-based foundation as "promising indeed" does not meet those responsibilities. PMID- 15801338 TI - Composite resin fluorescence. PMID- 15801340 TI - Dentinal bonding reaches the root canal system. AB - Several factors can contribute to the achievement of success with endodontic therapy. After an effective microbial-control phase, an adequate canal and coronal filling will guarantee a high probability of success. Gutta-percha has for many years been widely used as a solid material in root fillings associated with different types of sealers. Even associated with a sealer, this material it is not capable of preventing leakage, as has been shown in many studies. In fact, because of gutta-percha's limitations, the seal of a coronal restoration may be as important as the gutta-percha fill in preventing reinfection of the root canal. Although sealers can form close adhesion to the root canal wall, none is able to bond to the gutta-percha core material. Upon setting, the sealer pulls away from the gutta-percha core, leaving a gap through which bacteria may pass. This article describes a new thermoplastic, synthetic root canal filling material, whose design is based on polyester chemistry, that looks and handles like gutta-percha. It is used in the same manner as most bonding systems. After the usual preparation of the root canal, a self-etch primer is used to condition the canal walls and prepare them for bonding to the resin. The resin sealant is introduced in the root canal. It bonds to the primer and to the resin core material; thus, a "monoblock" is formed without the gaps typical in gutta-percha fillings. Studies have shown that leakage of bacteria with this material is significantly reduced compared with gutta-percha. PMID- 15801339 TI - Clinical strategies for success in proximoincisal composite restorations. Part I: Understanding color and composite selection. AB - The restoration of proximoincisal (Class IV) defects with direct resin-based composites requires attention to many technical and artistic details. This article is the first of a series of two articles that aim at presenting clinical strategies for optimal success when direct resin-based composites are used for the restoration of moderate or large proximoincisal defects. Concepts of natural anatomy, color as it relates to dental structures, and composite selection are discussed in this article and are illustrated with a preclinical exercise and two clinical cases in which these concepts are applied. PMID- 15801341 TI - Does the wear resistance of packable composite equal that of dental amalgam? AB - BACKGROUND: There is little evidence that packable composites are sufficiently wear resistant to be used as an alternative to amalgam. PURPOSE: The purpose of this study was to evaluate wear rates of packable composites compared with hybrid resin composites and amalgams by an in vitro wear test. MATERIALS AND METHODS: The following composites were used: three packable composites (SureFil, Dentsply/Caulk, Milford, DE, USA; Alert, Pentron Clinical Technologies, Wallingford, CT, USA; and Solitaire, Heraeus Kulzer, Wehrheim, Germany), two hybrid resin composites (TPH Spectrum, Dentsply/Caulk; and Pyramid enamel, Bisco Inc., Schaumburg, IL, USA), and two amalgams (Tytin, Kerr Manufacturing Co., Romulus, MI, USA; and Dispersalloy, Dentsply/ Caulk). Cylindrical Class I cavities prepared on occlusally flattened, extracted human molars were restored with respective materials according to the manufacturers' instructions. Generalized, localized, and antagonistic enamel wear tests were carried out by a University of Alabama at Birmingham (UAB) wear simulator according to previously reported methods. Seven specimens were tested for each group, and the wear depths were measured on profilometric tracings. The data for each wear mode were independently analyzed by one-way analysis of variance and Fisher's exact test (p < or = .05). RESULTS: The generalized wear values for SureFil (7.0 +/- 3.5 microm), Alert (8.6 +/- 1.8 microm), and Pyramid (3.9 +/- 0.5 microm) were not statistically different from those of amalgam materials (Tytin 5.8 +/- 0.7 microm, Dispersalloy 6.0 +/- 0.9 microm) but were different from those of Solitaire (23.9 +/- 2.6 microm) and TPH (30.6 +/- 5.5 microm). The localized wear values for SureFil (19.8 +/- 14.2 microm) and Alert (28.0 +/- 1.6 microm) were significantly smaller than for all other materials. For antagonistic enamel wear, Solitaire exhibited a minimal value (3.4 +/- 0.9 microm), whereas values of SureFil (12.6 +/- 5.6 microm) and Alert (12.0 +/- 6.6 microm) were not statistically different from those of TPH (11.0 +/- 4.0 microm) and amalgams (Tytin 14.5 +/- 4.3 microm, Dispersalloy 7.8 +/- 3.3 microm). CONCLUSIONS: It can be concluded that SureFil and Alert packable composites possess similar wear resistance and abrasiveness to amalgam on the basis of the limitations of this study, which simulated 3 years of clinical wear. PMID- 15801342 TI - Comparison of effectiveness of two 10% carbamide peroxide tooth-bleaching systems using spectrophotometric measurements. AB - PURPOSE: The purpose of this study was to compare the color changes achieved with two commonly used bleaching systems on the basis of a spectrophotometric analysis. MATERIALS AND METHODS: Two commercially available 10% carbamide peroxide bleaching systems were used by a total of 48 individuals. Subjects who had intact natural maxillary anterior teeth void of any restorations or decay that had not been subjected to any prior bleaching were included. Opalescence 10% PF (Ultradent Products Inc., South Jordan, UT, USA) was used by 23 patients for 14 days, whereas Nite White Excel (Discus Dental, Culver City, CA, USA) was applied by 25 patients for the same duration. Tooth colors for the middle one third region of maxillary central incisors and canines were measured with a spectrophotometer prior to bleaching and after 14 days of bleaching. Color difference deltaE and color coordinates L* (lightness), a* (redness), and b* (yellowness) of CIELAB color system (developed by the Commission Internationale de l'Eclairage) were calculated. RESULTS: Mean deltaE values for Opalescence ranged from 5.03 to 8.92 and from 5.84 to 9.61 for Nite White. The most significant factor of the color change was b* followed by L* and a*. L* values were higher after bleaching, whereas values for both a* and b* decreased. There were no significant differences between the two systems examined (p < .05). PMID- 15801343 TI - Safety of dental amalgam. PMID- 15801344 TI - Diastema closure. PMID- 15801345 TI - Preparing the older adult for surgery and anesthesia. AB - Life expectancy in the United States has increased significantly over the past 100 years. It has been suggested that by the year 2050, approximately 20% of the population may be over the age of 65. Many of these older adults will undergo surgery and anesthesia. Preadmission testing nurses need to be able to deal with the various challenges that accompany this group of patients. While many of the elderly are quite active, there are others with multiple disease processes that put them at an increased risk over the general population. It will be to the advantage of all involved if health care providers can develop a better understanding and an increased expertise in caring for the geriatric surgical patient. This article describes the importance of the preanesthetic nursing assessment and evaluation in helping to identify potential problems so that a plan of care to prevent those problems can be formulated and communicated to other members of the health care team. PMID- 15801346 TI - Geriatric anesthesia implications. AB - The geriatric patient will continue to pose challenges during the perianesthesia period. By age 80, the older adult has experienced many physiological and anatomical changes, many starting during the 4th decade of life. Changes in tissue mass and function in major organ systems demand special perianesthetic care planning to optimize patient outcomes. Understanding the physiology of aging will assist the perianesthesia nurse in planning perianesthesia care for their patients. Consequently, emphasis should be placed on cautious preanesthesia screening and evaluation, so that astute diagnosis and treatment of comorbid diseases are thoughtfully considered in relationship to the type of anesthesia when caring for the postanesthesia patient. For this article the term geriatric refers to those persons 65 years of age or older. PMID- 15801347 TI - Perianesthesia challenges in geriatric pain management. AB - The perianesthesia nurse is continually challenged to provide safe and effective pain management. This task becomes more difficult when working with an aging population. Success begins with understanding the challenges in controlling surgical pain and in communicating information regarding pain management prior to the surgery. Sharing of information between the patient, nurse, and other members of the health care team is essential. To provide quality pain care for geriatric patients, who are at greater risk for developing potentially life-threatening side effects of commonly used analgesics, the perianesthesia nurse must be knowledgeable about factors that affect pain management in this population. Common factors affecting pain control in the older adult patient include misconceptions regarding use and effects of analgesics, preexisting cognitive impairment, impaired communication, cultural differences between the nurse and the patient, and physiologic changes in aging that affect how drugs are metabolized. PMID- 15801348 TI - When the trauma patient is elderly. AB - Each year thousands of older adults will suffer trauma injuries. Geriatric patients who sustain serious injuries are at risk for increased morbidity and mortality. Preexisting medical conditions can dramatically impact every aspect of care in older patients. As a direct response to the stress of injury, reserve functioning can be put to the test. Vital signs may initially appear normal. However, compensatory mechanisms in older individuals may not be predictable in response to physiological needs. Perianesthesia nurses caring for older trauma patients should be familiar with the anatomical and physiological changes of aging, and have an awareness of the original mechanism of injury, coupled with the surgical procedure and anesthesia agents. Astute nursing assessments using critical thinking skills, together with effective nursing interventions, provide the framework for understanding and managing geriatric trauma patients in the perianesthesia care continuum. PMID- 15801349 TI - Discharge planning for the elderly ambulatory surgical patient. AB - Discharge planning for the elderly ambulatory surgery patient should begin as soon as the patient's procedure is booked. Successful discharge planning relies on 1) comprehensive preoperative assessment; 2) effective communication among the surgical facility's caregivers, the physician's office, the patient, and the family; 3) consideration of the patient's preoperative status; and 4) a strong patient and family education plan. The challenges of planning the aftercare are compounded by the physical, social, financial, cognitive, and sensory changes related to the normal aging process as well as to systemic diseases, which are more prevalent in the older population than in younger patients. The elderly patient's discharge plan should identify and address age-related barriers to communication, incorporate the patient's existing physical and medical condition, diminish the negative effects of social support challenges, and address environmental issues that can be improved to support recovery. The elderly population often deals with the imperfect: isolation from family or friends, aging bodies, hearing and visual loss, financial limitations, and emotional challenges. Although the nurse cannot reverse these challenges of old age, he or she can still make a difference by providing guidance and resources to blunt the potential complications of surgery and anesthesia. PMID- 15801350 TI - Surgical considerations in the elderly. AB - The number of older adults in America is rising every year. This increase in the elderly population will have a direct impact on health care delivery and perianesthesia nursing. Consequently, nurses need to increase their knowledge regarding the geriatric patient undergoing surgery. Areas of particular importance for nurses are preoperative assessment, medications taken by the elderly, preoperative instructions, intraoperative nursing care, postoperative nursing care, and discharge planning. Also of particular concern for the elderly surgical patient are nutrition, skin integrity, pain management, and promotion of sleep and comfort. The geriatric client requires extra time when being prepared for both elective and emergency surgery. A little planning goes a long way, and nurses need to enhance their communications skills to promote positive outcomes for the geriatric surgical patient. Caring for the elderly surgical patient can be a significant challenge, but if nurses work together as part of the perianesthesia/perioperative team, the experience can be extremely rewarding. PMID- 15801351 TI - Medication use in the elderly patient: focus on the perioperative/perianesthesia setting. AB - As the population ages, the use of multiple medications also increases. Polypharmacy (taking multiple drugs at a time) presents concerns to the perianesthesia nurse who is caring for the geriatric patient. The pharmacokinetics and pharmacodynamics of drugs are often altered in older adult patients. Adverse drug reactions and drug interactions occur more often in geriatric patients than in younger patients. For these reasons, the benefits and risks of multiple medications and the administration of certain types of drugs must be carefully considered in the elderly patient. The selection of any medication should be individually based on the benefits and risks. Adverse drug reactions play a significant role in hospitalization for the general population, and the elderly are more susceptible to these. These drug reactions often contribute to significant morbidity as well as mortality. Medications need to be considered carefully in the older adult patient, but perhaps more so in the perioperative/perianesthesia period. Drug interactions are diverse. The type of anesthesia may influence the patient's outcome, depending on the medications the patient is currently taking. The patient's response to the stress of surgery is also affected by individual medical conditions as well as medications the patient is currently receiving Polypharmacy, inappropriate medications, adverse drug reactions, drug-disease issues, and drug interactions in the geriatric population are concerns in the perioperative/perianesthesia setting. PMID- 15801352 TI - Community resources for the elderly. AB - There are many, many community resources available for the elderly. Your local Yellow Pages Directory is very helpful, and by surfing the Internet, you can access far more information than is possible to read. Physicians and hospital based case workers also offer a personal and reassuring touch. PMID- 15801353 TI - Issues of informed consent in the geriatric population. PMID- 15801354 TI - Restraints use and falls prevention. AB - The Health Care Financing Administration defines physical restraints as "any manual method or physical or mechanical device, material, or equipment attached or adjacent to the patient that the individual cannot remove easily which restricts freedom of movement or normal access to one's body. Restraints have the potential to produce serious outcomes, including physical or psychological harm, loss of dignity, violation of a patient's rights, and possibly death. Health care providers need to identify opportunities to decrease the risks associated with the use of restraints through preventive strategies, innovative alternatives, and process improvements to help focus on the patient's overall well-being, health, and safety. PMID- 15801355 TI - I am back! Thanks for being here for me!!! PMID- 15801356 TI - The melodious bell: the joy of work. PMID- 15801357 TI - Progress in the use of helical CT for imaging urinary calculi. AB - Helical CT has become the preferred method to diagnose urinary calculi in patients presenting with abdominal or flank pain. Recent in vitro studies have shown that CT also can display the internal structure in stones with remarkable detail. Because some stones respond better to SWL than others, knowing stone structure at diagnosis could be helpful in choosing among treatment options. This paper examines the potential for CT to be used in this way. Older CT technology proved to be problematic, in that all studies using low-resolution CT will suffer from an artifact in which stone size affects apparent CT attenuation values. Thus, the observation that stones with low measured CT attenuation break more easily than stones with high attenuation could be attributable entirely to an artifact of stone size. Most stones are composed of more than one mineral, and heterogeneity of composition may contribute to variability in stone response to SWL. Older technology is not useful in evaluating stone composition, but current and emerging CT machines have sufficient resolution to determine the composition and structure of stones inside the patient, provided proper viewing windows are used. Continuing improvement in image resolution in helical CT promises to provide information about stone composition and structure that will ultimately lead to better care for patients with stone disease. PMID- 15801358 TI - Efficacy of the Duet lithotripter using two energy sources for stone fragmentation by shockwaves: an in vitro study. AB - PURPOSE: To evaluate the efficacy of the Duet lithotripter's novel design of two independent spark-plug generator/reflector systems focused at a common F2. The apparatus allows either simultaneous delivery of shockwaves from both generators (resulting in a per-shock energy delivery at F2 equal to that delivered by its single generator at about 24 kV), alternating (between the two generators), or single-generator delivery of shockwaves at various energy levels and rates. MATERIALS AND METHODS: Eighty-five phantom gypsum stones (volume 786 mm3 each) were placed in a net-like basket and immersed in a specially designed waterbath coupled with the Duet lithotripter (Direx Medical Systems Ltd., Petach Tikva, Israel). Shockwaves were delivered at rates of either 60 or 120 per minute and at intensities of 16 or 22.8 kV (electrohydraulic). Energy was delivered either separately from each generator, in an alternating mode, or simultaneously from both generators. The number of shocks required to fragment the stones sufficiently to allow all of the pieces to fall through the basket holes (complete fragmentation) was recorded. RESULTS: The number of shocks required for complete fragmentation in the alternate mode (120 shocks/min, each generator rate 60/min; 22.8kV) was lower than with the single generator, 112 +/- 19 v 134 +/- 18 (at a rate of 120/min; 22.8 kV). The simultaneous mode of dual generator shockwave delivery was more effective than the traditional single generator (114 +/- 28 shocks at a rate of 120/min, 16 kV v 159 +/- 40 shocks at a rate 120/min; 22.8kV). CONCLUSION: The Duet lithotripter is more effective when used in a simultaneous or alternating mode than is the classical single mode of shock delivery, with the added benefit of shorter treatment time. PMID- 15801359 TI - Radial dilation force of tipless and helical stone baskets. AB - PURPOSE: To evaluate one aspect of tipless and helical stone basket function that is critical for ureteral stone extraction: the radial-dilation force. MATERIALS AND METHODS: Nine commercially available tipless baskets and five commercially available helical stone baskets were tested. Two Teflon blocks were positioned with the lower block sitting on a digital scale and the upper block secured to a plastic frame and base. A 0.01-inch gap was maintained between the blocks using a digital micrometer. Alignment pins secured the position of the lower block in relation to the upper block. A 4-mm cylindrical hole was drilled through the center of the block interface, and each basket was passed through the hole and opened to its fully extended length. The basket was then slowly retracted through the hole, and the maximum force reading was recorded. Twenty repetitions were performed for each basket. RESULTS: Of the tipless baskets > or =3.0F, the Cook N Circle 3.2F provided the best radial dilation (24.7 +/- 0.4 g). For tipless baskets <3.0F, the Sacred Heart Vantage 2.4F provided the best radial dilation (19.6 +/- 0.8 g). Of the helical baskets, the Sacred Heart Hercules provided the most radial dilation (102 +/- 12.1 g) followed by the Cook N-Force (71.8 +/- 4.3 g). CONCLUSION: The radial-dilation force of tipless and helical stone baskets differs significantly among baskets and may impact stone extraction performance in the ureter. PMID- 15801360 TI - Retroperitoneoscopic pyeloplasty for ureteropelvic junction obstruction. AB - BACKGROUND AND PURPOSE: Retroperitoneoscopy is an established procedure for renal surgery. We evaluated our results with retroperitoneoscopic pyeloplasty for ureteropelvic junction (UPJ) obstruction. PATIENTS AND METHODS: In 14 female and 10 male patients, a retroperitoneoscopic pyeloplasty was performed (13 right/11 left). Four patients had previously had an endopyelotomy. The operation was performed using standard retroperitoneoscopic technique with the patient in a flank position. After preparation of the ureter and renal pelvis, the UPJ was resected in 22 patients in an Anderson-Hynes pyeloplasty. Twenty of these patients had a crossing vessel. The other two patients, who had small renal pelves, were operated on with a Fenger pyeloplasty. In all patients a 7F double-J stent was placed. RESULTS: The mean operative time was 189 minutes (range 70-360 minutes), and the average blood loss was 110 mL (range 50-400 mL). There were no intraoperative complications, although one patient with adhesions and scarring after previous endopyelotomy had to be converted to open surgery. The transurethral catheter was left for 7 days in the first 10 cases and for 4 days in the 14 subsequent patients. The hospitalization time was 9.7 and 7.5 days, respectively. The only postoperative complication was a urinoma, which was punctured. The double-J catheter was removed after an average of 4.6 weeks (range 4-8 weeks). Intravenous urography 6 weeks later showed no obstruction. The mean follow-up time was 11.5 months (range 1-24 months) with no signs of obstruction on ultrasonography. CONCLUSIONS: Retroperitoneoscopic pyeloplasty for UPJ obstruction is a safe and effective procedure. Our short-term results are similar to those of open pyeloplasty with the advantage of a minimally invasive approach. PMID- 15801361 TI - Endopyelotomy in childhood: our experience with 37 patients. AB - PURPOSE: To evaluate our experience and results with endopyelotomy in the pediatric population. PATIENTS AND METHODS: Between 1990 and 2002, we performed percutaneous antegrade endopyelotomy under general anesthesia in 37 children because of ureteropelvic junction (UPJ) stricture. The youngest patient was 4.5 years and the oldest 17 years at the time of the procedure (mean age 11.5 years). One patient had bilateral stenosis; the two sides were operated on separately. After insertion of a 4F ureteral catheter and filling the collecting system with colored contrast material, a middle calix was punctured under fluoroscopic control. The tunnel was dilated to 26F by telescopic metal dilators. After insertion of a 0.035-inch gidewire through the UPJ, all its layers were cut by a cold knife in the dorsolateral direction so that the periureteral fatty tissue could be seen. Finally, the ureteral wound was stented by a 6F to 12F transrenal drain or a double-J catheter, which was removed after 6 weeks. RESULTS: Among the 37 patients, the procedure had to be repeated in 1 because the transrenal drain stenting the UPJ slid back to the renal pelvis. We had to perform open pyeloplasty or nephrectomy in two patients because of bleeding or failed procedure. The average postoperative hospital stay was 6 days. Comparison of the preoperative intravenous urograms with studies performed 1 year after endopyelotomy showed an overall success rate of 89%. All patients are without complaints at the moment. CONCLUSIONS: In experienced hands, endopyelotomy is a safe and effective method for the treatment of UPJ stricture, not only in the adult, but also in the pediatric, population. PMID- 15801362 TI - First clinical experience with new transurethral bipolar prostate electrosurgery resection system: controlled tissue ablation (coblation technology). AB - BACKGROUND AND PURPOSE: Bipolar transurethral resection of the prostate (TURP) can allow transurethral debulking of the adenoma with a lower potential for complications from systemic absorption of hypotonic irrigant. We evaluated a new bipolar double-loop resection system with a dedicated controlled ablation (Coblation) generator in this context. PATIENTS AND METHODS: In our series of 36 patients, 32 underwent transurethral bipolar loop resection, of whom 12 had prostates >50 cc (range for entire series 30-126 cc) on transrectal ultrasonography. Ten men had indwelling catheters because of urinary retention. The four other patients underwent transurethral resection of bladder tumors. After an initial learning curve of eight TURPs, some minor technical modifications were made, both to the bipolar-loop resection technique and to the hardware. The modified Vista Coblation system was subsequently evaluated by 18 other urologists in 17 other centers throughout the U.S. RESULTS: The median resected dry weight was 22 g, and the median operating-room time was 48 minutes, giving a median resection rate of approximately 1 g every 2 minutes. Two of the eight men in our learning curve required treatment for delayed bleeding problems (clot obstruction of catheter), and one of these received a blood transfusion. Transfusion was not required subsequently regardless of prostate size, even in men with larger glands. One of the patients treated for bladder tumor had a minor perforation of the posterior bladder wall, which healed with conservative management. In the multicenter experience in the U.S., there were 59 evaluable patients. The median resected weight was 23 g, and the median resection time was 55 minutes. Two men required conversion to monopolar roller coagulation for intraoperative bleeding. Most patients (80%) were discharged from hospital within 24 hours. No adverse neuromuscular stimulation occurred. CONCLUSION: This first single-center and international multicenter experience with Coblation technology for bipolar double-loop saline TURP confirms the feasibility and safety of this procedure. Subjective evaluation showed the four most important perceived benefits of bipolar over monopolar TURP to be smoother cutting action, elimination of TUR syndrome, less tissue burning (and no smell), and reduced bleeding. The results of randomized studies with this technology are awaited, while its role in bladder tumor resection remains unclear. PMID- 15801363 TI - Effects of intraprostatic and periprostatic injections of mepivacaine epinephrine on intraprostatic blood flow during transurethral microwave thermotherapy: correlation with [15O]H2O-PET. AB - BACKGROUND AND PURPOSE: Experiences with the ProstaLund Feedback Treatment (PLFT) with the CoreTherm device and results from a previous positron emission tomography (PET) study suggest that the intraprostatic blood flow increases considerably during treatment in response to heat exposure. Early results with intraprostatic injection of mepivacaine epinephrine prior to PLFT have indicated greater patient comfort during treatment and shorter treatment time secondary to lower intraprostatic blood flow. In this pilot study, the effect of intraprostatic injection of mepivacaine epinephrine on intraprostatic blood flow before and during PLFT was evaluated by PET using [15O]H2O. PATIENTS AND METHODS: In four patients scheduled for PLFT, a baseline value of the intraprostatic blood flow was established using [15O]H2O-PET. Thereafter, intraprostatic injections of mepivacaine epinephrine were given using a prototype of the Schelin Catheter. In two of the patients, PET was performed immediately after the mepivacaine epinephrine injections and 10 and 24 minutes after the start of PLFT. To reduce the risk of wash-out of the drug, the next two patients were examined 7 and 17 minutes after the start of PLFT but not in connection with the anethetic injection. RESULTS: In patients 1 and 2, mepivacaine epinephrine decreased the prostatic blood flow. During PLFT, there was a slight increase in blood flow in patient 1 and a more pronounced increase in patient 2. In patient 3, the blood flow during treatment was almost unchanged, while it decreased in patient 4. CONCLUSIONS: Intraprostatic injection of mepivacaine epinephrine may reduce, or even eliminate, the increase in blood flow that is usually seen during PLFT. The vague effect seen in patients 1 and 2 may be explained by wash-out of the drug. PMID- 15801364 TI - Simultaneous holmium laser enucleation of prostate and upper-tract endourologic stone procedures. AB - BACKGROUND AND PURPOSE: The holmium laser is a versatile urologic tool. Its unique cutting and coagulating properties allow multiple procedures, such as stone fragmentation and laser enucleation of the prostate (HoLEP), to be performed with a single set-up. This paper reviews our experience with simultaneous HoLEP and endourologic upper-tract stone procedures. PATIENTS AND METHODS: We retrospectively reviewed all 11 patients (12 renal units) treated with the combined approach of HoLEP and an upper-tract endourologic procedure for stone disease. All patients initially underwent HoLEP, followed immediately by percutaneous nephrolithotomy (PCNL)(N=2), ureteroscopy (URS)(N=8), or both (N=1). The mean stone diameter was 34.7 mm for PCNL and 6.7 mm for URS. Outcome data and complications were recorded. RESULTS: The mean preoperative and postoperative American Urological Association Symptom Scores were 21.8 and 6.3, respectively. The mean prostatic specimen weight was 118 g (range 21-376 g), and the mean hospital stay was 1.4 days. For 10 patients with available prostatic specific antigen (PSA) data, the mean preoperative and postoperative values were 6.2 and 0.9 ng/mL, respectively. All patients were catheter free at discharge. All three PCNL patients were rendered stone free; all four URS patients who had radiographic follow-up were stone free. There were no short- or long-term complications, and no transfusions were necessary. CONCLUSIONS: Simultaneous HoLEP and laser lithotripsy are feasible, emphasizing the unique capabilities of the holmium laser. The hemostasis offered by HoLEP allows safe removal of prostatic tissue and urinary calculi, obviating multiple procedures. PMID- 15801365 TI - Photoselective KTP laser vaporization of the prostate: first experiences with 65 procedures. AB - PURPOSE: To study the feasibility and efficacy of 80 W potassium-titanyl phosphate (KTP) laser vaporization (GreenLight PV; Laserscope) of the prostate in patients suffering from voiding dysfunction secondary to benign prostatic hyperplasia (BPH) or known locally advanced prostate cancer (CaP). PATIENTS AND METHODS: Sixty-five patients with symptomatic BPH (N=57) or obstructive voiding secondary to CaP (N=8) with a mean age of 70 +/- 10 years (range 46-93 years) underwent photoselective 80 W KTP laser vaporization of the prostate. All consecutive patients, including 34 with a history of chronic urinary retention or indwelling catheter, were enrolled. Prostate specific antigen (PSA) values, prostate volume as measured by transrectal ultrasonography, urinary peak flow measurement (Qmax), postvoiding residual volume (PVR) measured transvesically, and International Prostate Symptom Score (IPSS) were assessed preoperatively, on the day of discharge, and 1 month and 3 months postoperatively. The mean preoperative prostate volume was 49 +/- 32 cc (range 15-250 cc). RESULTS: In all 65 patients, KTP laser vaporization was performed successfully, with a mean operating time of 57 +/- 25 minutes (range 10-160 minutes). No major complication occurred intraoperatively or postoperatively, and no transfusions were necessary. All patients were catheter free after 1 month. At 1 month and 3 months, the urinary peak flow had increased from 7.7 +/- 2.8 mL/sec preoperatively to 20.9 +/ 11.6 mL/sec (+171%) and 18.2 +/- 6.3 mL/sec (+136%), respectively. The IPSS decreased from 18.5 +/- 6.7 to 9.2 +/- 7.7 (-50%) and 7.2 +/- 5.9 (-61%) at 1 and 3 months, respectively. CONCLUSIONS: A 80 W KTP laser vaporization of the prostate technique instantly removes obstructive tissue. A transurethral resection-like visible cavity is the endpoint. Immediate symptom relief is achieved in a truly minimally invasive way with a very low postoperative complication rate within 3 months' follow-up. PMID- 15801366 TI - Smooth-muscle regeneration after electrosurgical endopyelotomy in a porcine model as confirmed by electron microscopy. AB - BACKGROUND AND PURPOSE: Endopyelotomy is the preferred treatment for ureteropelvic junction (UPJ) obstruction because of its short operating time, limited morbidity, fast recovery, and reasonable efficacy. We used tissue and immunohistochemistry staining and electron microscopy to look at the muscle regeneration following an endopyelotomy incision in a porcine model. MATERIALS AND METHODS: Bilateral electrosurgical endopyelotomy was performed in six domestic pigs with placement of 7F 20-cm Percuflex double-J stents for up to 4 weeks, and urinary tracts were harvested at 3 or 5 months. Specimen evaluation included tissue staining with hematoxylin-eosin, Masson's trichrome, and Verhoeff's iodine and Van Gieson solution; histochemical staining for smooth muscle actin, desmin and myosin staining, and electron microscopy. Each specimen was assigned a "healing" score of 0 (normal) 1 (slight changes), 2 (mild changes), or 3 (severe changes). The fibrosis score was based on six factors: muscle layer fibrosis, lamina propria fibrosis, amount of granulation tissue present, new deposits of collagen, fibrosis in the periureteral fat, and presence of myofibroblasts. The muscles were characterized with immunohistochemistry and electron microscopy. RESULTS: At both 3 and 5 months, the urothelium was healed, and the lamina propria was healed with focal loss. By 3 months, smooth-muscle bundles bridged the defect, and by 5 months, the whole defect was covered. Smooth muscle cells were evident by electron microscopy by 3 months, and actin and myosin could be detected by immunohistochemistry. Desmin-positive cells accounted for 50% of the population at 3 months and 40% at 5 months. The regenerated smooth muscle bundles were oriented in different directions and intermingled with fibrous tissue. They could be distinguished easily from normal ureter under the microscope. CONCLUSION: Verifiable, functional smooth-muscle bundles bridge the endopyelotomy defect by 3 months, as confirmed by immunohistochemistry staining and electron microscopy. PMID- 15801367 TI - Opening keynote address: the political landscape and how it impacts healthcare. PMID- 15801368 TI - Epidemic of care. PMID- 15801369 TI - Healthcare disruption and the "perfect storm". PMID- 15801370 TI - Healthcare expert panel: health cost drivers and consumer empowerment. PMID- 15801371 TI - Health information technology and e-prescribing: a convergence of payor and provider interests. PMID- 15801372 TI - "Personalized" medicine: changing the practice and economics of healthcare. PMID- 15801373 TI - Pharmaceutical expert panel: taming the drug cost monster. PMID- 15801374 TI - Closing keynote address: American vital signs 2004. PMID- 15801375 TI - Conflicts, defenses and the stability of narcissistic personality features. PMID- 15801376 TI - Meeting the needs of parentally bereaved children: a framework for child-centered parenting. AB - This article describes the development and deployment of a framework for measuring parenting capacities in the context of bereavement. Grounded theoretical analysis of interviews with a community sample of 41 bereaved spouses with school-aged children elicited a set of nine bereavement-specific parenting tasks. A corollary coding system (covering all nine parenting tasks) was created to transform interview materials into quantitative data, thus permitting systematic empirical investigation of the parenting capacities of bereaved spouses. Parenting behaviors were coded on a 5-point scale ranging from least child-centered to most child-centered. Sex of surviving parent and circumstances of death proved to be significant mediating variables: mothers were more child centered than fathers, and parents surviving sudden deaths more child-centered than those surviving anticipated deaths. Lengthy illness was associated with less child-centered parenting. The more child-centered the parenting, the less symptomatic the child as measured by parent report (Child Behavior Checklist) and child self-report (Children's Depression Inventory, Revised Child Manifest Anxiety Scale). Child-centered parenting was associated with more positive and fewer negative perceptions of the surviving parent by the child as measured by the Parent Perception Inventory. Implications of findings are discussed. PMID- 15801377 TI - Brain environment interactions: stress, posttraumatic stress disorder, and the need for a postmortem brain collection. AB - Stress, especially the extreme stress of traumatic events, can alter both neurobiology and behavior. Such extreme environmental situations provide a useful model for understanding environmental influences on human biology and behavior. This paper will review some of the evidence of brain alterations that occur with exposure to environmental stress. This will include recent studies using neuroimaging and will address the need for histological confirmation of imaging study results. We will review the current scientific approaches to understanding brain environment interactions, and then make the case for the collection and study of postmortem brain tissue for the advancement of our understanding of the effects of environment on the brain. Creating a brain tissue collection specifically for the investigation of the effects of extreme environmental stressors fills a gap in the current research; it will provide another of the important pieces to the puzzle that constitutes the scientific investigation of negative effects of environmental exposures. Such a resource will facilitate new discoveries related to the psychiatric illnesses of acute stress disorder and posttraumatic stress disorder, and can enable scientists to correlate structural and functional imaging findings with tissue abnormalities, which is essential to validate the results of recent imaging studies. PMID- 15801378 TI - Toward a national PTSD brain bank. PMID- 15801379 TI - Understanding heterogeneous effects of trauma exposure: relevance to postmortem studies of PTSD. PMID- 15801380 TI - What's missing in posttraumatic stress disorder research? Studies of human postmortem tissue. PMID- 15801381 TI - The challenge of a PTSD brain bank. PMID- 15801382 TI - Neuropathology of stress: prospects and caveats. PMID- 15801383 TI - Brain environment interactions. PMID- 15801384 TI - The development of a posttraumatic stress disorder brain collection. PMID- 15801385 TI - Physical and cognitive activity and exercise for older adults: a review. AB - Age-related reduction in musculoskeletal, cardiovascular, and central nervous system resilience can result in wide-ranging limitations in adaptive capacity associated with negative outcomes such as cognitive decline, increased risk of cardiovascular disease, mobility problems, and increased incidence of debilitating falls. This article reviews the benefits of both cognitive and physical activity within the broad context of multiple system resilience in adult aging. Research on a unique form of combined physical/cognitive exercise, Tai Chi Chuan, is presented. The relationship between physiological and psychological gain associated with an activity intervention program is discussed in light of principles of rehabilitation, intervention compliance, subjective and objective gain, and the hypothesized value of combining physical exercise, cognitive exercise, and relaxation into a single program designed to promote resilience in older adults. PMID- 15801386 TI - Self-transcendence: conceptualization and measurement. AB - Self-transcendence has been hypothesized to be a critical component of wisdom (Curnow, 1999) and adaptation in later life (Tornstam, 1994). It reflects a decreasing reliance on externals for definition of the self, increasing interiority and spirituality, and a greater sense of connectedness with past and future generations. The Adult Self-Transcendence Inventory was administered to 351 individuals along with the NEO-FFI Personality Scale (McCrae & Costa, 1989). A principal axis factor analysis identified two factors: self-transcendence and alienation. The relationships between self-transcendence and neuroticism, openness to experience, extraversion, and agreeableness were significant, although modest, suggesting that self-transcendence cannot be accounted for in terms of positive personality traits alone. As expected, a multiple regression analysis indicated that self-transcendence was negatively related to neuroticism and positively related to meditation practice. The present study appears to lend support to the construct of self-transcendence. PMID- 15801387 TI - Balancing between contradictions: the meaning of interaction with people suffering from dementia and "behavioral disturbances". AB - Interacting with people who suffer from dementia poses a challenge for care providers, and the presence of behavioral disturbances adds a further complication. Our article is based on the assumption that behavioral disturbances are meaningful expressions of experiences. Six narrative interviews were conducted with care providers with the aim of illuminating the meaning of interaction with people suffering from dementia and behavioral disturbances. The interviews were tape-recorded, transcribed into text, and interpreted using a phenomenological hermeneutic methodology. The findings indicate that interacting with people with dementia and behavioral disturbances, as narrated by care providers, means balancing between contradictions concerning meeting the person in my versus her/his world, feeling powerless versus capable, and feeling rejected versus accepted. Interaction involves being at various positions along these continua at different points in time. Furthermore, it means facing ethical dilemmas concerning doing good for the individual or the collective. This is interpreted as a dialectic process and is reflected on in light of Hegel's reasoning about the struggle between the master and the slave. PMID- 15801388 TI - Age related differences in the strategies used by middle aged adults to solve a block design task. AB - In the present study, it was proposed to investigate the effects of aging on the strategies used to solve a block design task and to establish whether these strategies may be associated with differential patterns of ability. Two groups of subjects, 30 young adults (aged 20-35 years) and 30 middle-aged adults (aged 45 60 years) were set a computer version of the Kohs task and a battery of tests. An age-related decrease in fluid intelligence (Gf) and visual-spatial ability (Gv) was observed, along with the fact that most of the older subjects used a global strategy rather than a synthetic one. On the other hand, while continuing to use strategies of the analytic type, the older subjects looked more frequently at the model and scored high on crystallized intelligence (Gc). These findings are discussed from two different points of view: the theory of hierarchical stimuli and the hypothesis that metacognitive ability, which is thought to rely on Gc, may increase with age, and thus compensate for the loss of Gf and Gv. PMID- 15801389 TI - [Acting against the rise of cannabis consumption]. PMID- 15801390 TI - [X fragile syndrome; how to make a precocious diagnostic]. AB - Fragile X syndrome diagnosis is not so easy to make in the first years. Morphological anomalies may be subtle, with high forehead, long face and large ears with soft cartilage. Autistic behaviour is frequent, with hand flapping, lone playing, eye contact avoidance and no language. Early diagnosis of X fragile syndrome is needed, as it is a genetic disease, with a recurrence risk in the family. Early diagnosis is also useful for the care of these children, with psychological and educational special management. PMID- 15801391 TI - [Cannabis: the approach must be scientific]. PMID- 15801392 TI - [Epidemiology of cannabis use]. AB - Cannabis is the most consumed illicit drug in France. Its use concerns especially men, in all social groups. It is mainly consumed by young adults and teenagers. Over the last ten years the consumption of cannabis strongly increased amongst French young people, the rise has been even stronger for regular use. Prevalence use in France has now reached the highest European levels. Cannabis use can lead to a treatment demand and a research will soon allow to screen problematic users in general population in France. The part of the cannabis in the activity of specialised centres for drug addicts progresses. Due to a lack of indicators, the sanitary impact is not well-informed; nevertheless there are presumptions that cannabis is involved in road accidents, cancers and psychiatric pathologies. The cannabis constitutes the main part of the arrests for drug use. PMID- 15801393 TI - [Acute and chronic cognitive disorders caused by cannabis use]. AB - Cannabis is currently the most commonly used illegal psychoactive substance amongst young people aged between 15 and 24, and it seems that 5% of this age group is addicted to it. Many research teams focused particularly on the cognitive disorders caused by cannabis use. Amongst the cognitive functions considered, memory-related, attention-related, psychomotor and motivation-related functions were proved deteriorated by acute and chronic cannabis use; a very important point, especially among teenagers, as possible alteration at the social and academic level could be the outcome. However studies on long-term and persistent cognitive effects haven't provided convergent conclusions. Methodological differences could partly affect these observations. Therefore it seems necessary to develop studies with larger samples. PMID- 15801394 TI - [Related, induced and associated psychiatric disorders to cannabis]. AB - Cannabis disorders, according to the DSM-IV and the ICD-10 criteria, include cannabis intoxication, cannabis abuse, cannabis dependence, and cannabis-related disorders (anxiety disorders, psychotic disorders, cannabis intoxication delirium). Although cannabis withdrawal syndrome has clinical importance, it is not included in these classifications. The amotivational syndrome remains controversial. The psychiatric disorders related to cannabis use are anxiety disorders, depressive disorders and psychotic disorders. Cannabis use could be closely linked with the neurobiology of schizophrenia. As the other psychoactive substances, cannabis use worsens the psychiatric outcomes and is associated with poorer treatment compliance. PMID- 15801395 TI - [Psychopathology of the cannabis user teenager]. AB - Over the past 10 years, cannabis use among adolescents has considerably increased. It became a major issue in public health. Today, the cannabis adolescents smoke is more concentrated in tetrahydroxycannabinol. Most of young people are using cannabis to deal with their psychological problems. In a way, cannabis use is more therapeutic than recreative. Substance abuse undercovers adolescent identity fragility. This fragility could lead to depression. The adolescent, by trying to manage this depression with cannabis use, jeopardize his individualisation process. Addiction has a specific function in adolescent psyche. It regulates the interpersonal relationship which is been considered as a treat by the adolescent. By this way, he's trying to avoid intrusion and abandonment anxiety. PMID- 15801396 TI - [Adverse effects of marijuana]. AB - When admitted in an emergency unit, young patients often present acute neurological effects of smoked marijuana. Other chronic adverse effects of marijuana are probably underestimated: postural syncope, arteritis, chronic bronchitis, amnesia. Marijuana may trigger a myocardial infarction and have a vasospastic effect. Marijuana has impairing effects on driving ability. Smoked marijuana is a potential respiratory tract carcinogen. PMID- 15801397 TI - [Use of cannabis: screening strategy and evaluation of severity factors]. AB - Cannabis consumption occupies a front place in the field of addictions in France, not only by the levels of consumption but also by the related health harms induced with a growing potentially problematic use of cannabis among young men (14% of those aged 14 to 18 in 2001). Therefore, it is not only an educational problem but also a public health issue. Harmful cannabis use must be systematically researched by history taking on the consumption of tobacco, alcohol and drugs as an integrated part of the patient's social history. The diagnostic of cannabis abuse is made by a clinical evaluation of harmful use, the frequency and quantity of consumption, the search for complications. The practitioner must also evaluate the factors of severity (modalities of consumption and parameters predictive of poor outcome like precocity of first experiences, auto-therapeutic use and the various types of individual and social vulnerability: personality characteristics, psychiatric pathology, family characteristics and social environment). The physician may also be helped by drug screening diagnostic and evaluation instruments like validated autoquestionnaires. Urinary screening can be used especially in public health or forensic medicine fields. Some situational risks must be known while cannabis intoxicated: pregnant women, mentally ill, driving, working. Finally, professionals must know that present situation is very different from what we knew in the seventies and that they have to cope with this fast moving issue. PMID- 15801398 TI - [Treatment of cannabis dependence]. AB - Cannabis is the most regularly used illicit drug in the world. Yet, until recently, there existed no therapeutic intervention for the users of this product. This was due to the fact that cannabis has always been considered a substance that doesn't lead to dependence and thus didn't necessitate a management plan for quitting. The results of the first studies in the domain of cannabis dependence suggest that this pathology responds to the same types of treatments for alcohol and tobacco dependence. PMID- 15801399 TI - [Cannabis in Medline]. PMID- 15801400 TI - [Cannabis: user or consumer at risk? Decisional balance]. PMID- 15801401 TI - [ECN: a new cycle of the References universitaires]. PMID- 15801402 TI - [Erectile dysfunction]. PMID- 15801403 TI - [Radiculopathy and carpal tunnel syndrome]. PMID- 15801404 TI - [Elevated serum creatinine]. PMID- 15801405 TI - [Limb pain and pain in extremities]. PMID- 15801406 TI - [Raynaud's phenomenon]. PMID- 15801407 TI - [In the 19th century, women attacking medicine]. PMID- 15801408 TI - Pore morphologies in disordered nanoporous thin films. AB - Materials with nanometer size heterogeneities are commonplace in the chemical and biological sciences (e.g, polymer blends, microemulsions, gels) and often exhibit complex morphologies. Although this morphology has a dramatic effect on the materials' properties, it is often difficult to accurately characterize. We describe a method, using small-angle X-ray scattering data, of generating representative three-dimensional morphologies of isotropic two-phase materials where the morphology is disordered, and we apply this to thin films containing nanometer sized pores with a range of porosities (4-44%). These representations provide a visualization of the pore morphology, give the pore size scale and extent of interconnection, and permit the determination of the transitions from closed pore to interconnected pores to bicontinuous morphology. PMID- 15801409 TI - Metallic contact formation for molecular electronics: interactions between vapor deposited metals and self-assembled monolayers of conjugated mono- and dithiols. AB - We present grazing-incidence Fourier transform infrared and AFM data of Au, Al, and Ti vapor-deposited onto self-assembled monolayers (SAMs) of conjugated mono- and dithiols. SAMs of 4,4'''-dimercapto-p-quaterphenyl, 4,4"-dimercapto-p terphenyl, and 4,4'-dimercapto-p-biphenyl have reactive thiols at the SAM/vacuum interface that interact with vapor-deposited Au or Al atoms, preventing metal penetration. Conjugated monothiols lack such metal blocking groups, and metals (Au, Al) can penetrate into their SAMs. Vapor deposition of Ti onto conjugated mono- and dithiol SAMs and onto hexadecanethiol SAMs destroys the monolayers. PMID- 15801410 TI - Phase behavior of the palmitic acid/palmitin system. A 2H NMR study. AB - The phase behavior of mixtures of palmitic acid (PA) and 1-monohexadecanoyl-rac glycerol, palmitin, was studied by phase contrast microscopy and deuterium solid state NMR. At pH 5, mixtures remained precipitated as lumps in solution. The NMR spectrum of the perdeuterated PA (PAd31) at 300 K exhibited a shape and quadrupolar splittings, deltav, characteristic of lipids embedded in a gel phase. The alkyl chains remained in a trans conformation with their long molecular axis oriented at about 15 degrees with respect to the bilayer normal. However, gauche defects were shown to occur at the end of the alkyl chain. At 330 K, the system underwent a phase transition to a hexagonal phase followed by an isotropic phase at 340 K. Upon cooling to 330 K, the spectrum in the hexagonal phase was oriented at 0 degrees showing that the cylinders were oriented with their long axis parallel to the field. Up to 11 positions (from 15) of PAd31 could be assigned. At pH 7 and 9 at room temperature, the mixtures were fully dispersed in a viscous solution of vesicles. The system underwent a phase transition at 320 K from a gel phase to a fluid phase with the bilayer normal oriented at 90 degrees with respect to the field. Analogous experiments performed with PA selectively labeled on carbon C2 allowed for the assignment of deltav for that position and suggested different conformations of the headgroup in the gel and fluid or hexagonal phases. The implications of these findings for the bio-availability of these fatty acids, in the understanding of the contribution of hydroxyl and carboxyl groups in the membrane formation, and for the production of simple self-oriented systems are discussed. PMID- 15801414 TI - Adsorption behavior of surface-chemically pure N-alkyl-N-(2 hydroxyethyl)aldonamides at the air/water interface. AB - N-alkyl-N-(2-hydroxyethyl)aldonamides (alkyl: n-C6H13, n-C8H17, n-C10H21, n C12H25, and n-C14H29) were obtained in the reaction of long-chain N-alkyl-N-(2 hydroxyethyl)amines with D-glucono-1,5-lactone and D-glucoheptono-1,4-lactone. The adsorption isotherms were obtained from surface tension measurements of aqueous solutions of surface-chemically pure surfactants. The experimental equilibrium surface tension versus concentration isotherms were evaluated by the Frumkin adsorption equation to get the adsorption parameters, namely, standard free energy of adsorption, deltaG(o)ad, saturation adsorption, gammainfinity minimum surface area demand per molecule adsorbed, Amin, and interaction parameter, Hs. The investigated functionalized alkylaldonamides show improved solubility in comparison with the corresponding sugar derivatives of the primary amines. The introduction of the -CHOH moiety into the saccharide headgroup causes a noticeable increase of the hydrophobic character of surfactant. The minimum surface area demand, Amin, is slightly greater for glucoheptonamides than for the corresponding gluconamides. The practically constant Amin value within the homologue series of the aldonamides indicates that the obtuse hydroxyethyl residue is the determining factor for the arrangement of the adsorbed surfactants in the interfacial layer. PMID- 15801415 TI - Cloud point of aqueous solutions of tetrabutylammonium dodecyl sulfate is a function of the concentration of counterions in the aqueous phase. AB - The cloud point of the surfactant tetrabutylammonium dodecyl sulfate is shown to be a function of the tetrabutylammonium counterion concentration in the aqueous phase whether the counterions are provided by the surfactant or both the surfactant and added tetrabutylammonium bromide. The micellized surfactant dissociates 17% of its counterions to aqueous phase. PMID- 15801417 TI - Mixed micellization of a nonionic-cationic surfactant system constituted by n octyl-beta-D-glucopyranoside/dodecyltrimethylammonium bromide/H2O. An electrochemical, thermodynamic, and spectroscopic study. AB - The mixed micelles constituted by a nonionic surfactant widely used in the biochemical field, n-octyl-beta-D-glucopyranoside, and a cationic surfactant with 12 carbon atoms on the hydrophobic tail, dodecyltrimethylammonium bromide, have been studied in aqueous solution, at 298.15 K, by means of conductivity, speed of sound, density, and fluorescence spectroscopy experiments. From these data, the monomeric and micellar phases of the mixed aggregates were fully analyzed through the determination of the total and partial critical micellar concentrations, the dissociation degree of the mixed micelle, the total and partial aggregation numbers, the apparent molar volumes and isentropic compressibilities, the hydration numbers, and the corresponding changes in these thermodynamic properties due to the mixed aggregation process. The experimental findings have been compared with those obtained with several theoretical models, some of them modified in this work to take into account the specific characteristics of the aggregates studied herein. PMID- 15801424 TI - Long-range order of organized oligonucleotide monolayers on Au(111) electrodes. AB - Oligonucleotides modified by a hexamethylene linker group adsorb on gold electrodes via Au-S bond formation. We have obtained novel data for adsorption of thiol-modified (HS) single-strand HS-10A and double-stranded HS-10AT oligonucleotides and for analogous thiol-free 10A (A = adenine) and 10T (T = thymine) nonspecifically adsorbed as reference molecules. Mercaptohexanol has served as a second reference molecule. The data are based on cyclic and differential pulse voltammetry, interfacial capacitance data, and in situ scanning tunneling microscopy (STM) directly in an aqueous buffer solution, with electrochemical potential control of both the sample electrode and the tip. All the data are based on single-crystal, atomically planar Au(111)-electrode surfaces. The high sensitivity of such surfaces provides accurate HS-10A and HS 10AT electrode coverages on the basis of the reductive desorption of the Au-S bond. The coverage is high and in keeping with dense monolayers of adsorbed HS 10A and HS-10AT in an upright or tilted orientation, with the oligonucleotide backbone repelled from the strongly negatively charged electrode surface. Adsorbed thiol-free 10A only gives a Au(111)-reconstruction peak, while 10T shows a subtle pattern involving pronounced voltammetric adsorption peaks indicative of both nonspecific adsorption via single thymine units and potential-dependent structural reorganization in the surface layer. In situ STM supports these findings at the molecular level. In situ STM of HS-10A discloses large, highly ordered domains at strongly negative sample potentials. Reversible domain formation and disordering could, moreover, be controlled by an electrochemical potential variation in the negative and positive directions, respectively. 10A and 10T did not form ordered adsorbate domains, substantiating that domain formation rests on adsorption of thiol-modified oligonucleotide adsorption in an upright or tilted orientation. The comprehensive, high-resolution information reported may hold prospects for single-molecule electronic conduction and molecular-scale mapping of oligonucleotide hybridization. PMID- 15801431 TI - Structural characterization of thin hydroxypropylcellulose films. X-ray reflectivity studies. AB - Thin solid films of hydroxypropylcellulose (HPC) have been investigated using synchrotron X-ray reflectivity. Evidence of preferential alignment of HPC molecules at the substrate surface is obtained. In the surface region the liquid crystalline domains ofHPC are preferentially oriented parallel to the substrate, whereas in the bulk they are mostly distributed randomly. Incorporation of colloidal particles in the film-substrate region destroys the preferential alignment. It is also found that in conditions of restricted geometry (very thin films), a minimum film thickness is required to produce lateral packing order perpendicular to the substrate surface. PMID- 15801437 TI - Multilayers of charged polypeptides as studied by in situ ellipsometry and quartz crystal microbalance with dissipation. AB - The buildup of poly(L-glutamic acid) (PGA) and poly(L-lysine) (PLL) multilayers on silica and titanium surfaces, with and without an initial layer of polyethyleneimine (PEI), was investigated and characterized by means of in situ ellipsometry and quartz crystal microbalance with dissipation. A two-regime buildup was found in all systems, where the length of the first slow-growing regime is dependent on the structure of the initial layers. In the second fast growing regime, the film thickness grows linearly while the mass increases more than linearly (close to exponentially) with the number of deposited layers. The film refractive indices as well as the water contents indicate that the film density changes as the multilayer film builds up. The change in film density was proposed to be due to polypeptides diffusing into the multilayer film as they attach. Furthermore, the use of PEI as the initial layer was found to induce a difference in the thickness increments for PGA and PLL. PMID- 15801444 TI - Molecularly imprinted TiO2 thin film by liquid phase deposition for the determination of L-glutamic acid. AB - For the first time, the feasibility of a molecularly imprinted liquid phase deposition (LPD) thin film has been demonstrated. Thin films of titanium oxide imprinted with L-glutamic acid were prepared by the LPD method on a gold-coated quartz crystal microbalance. The imprinted molecule could be removed upon treatment with immersion in deionized water. A sensor was developed on the basis of this method and showed good sensitivity, selectivity, and reproducibility to the template molecule. An equation was deduced to characterize the interaction between molecularly imprinted films and the template by virtue of Scatchard analysis. X-ray photoelectron spectroscopy was introduced to show the evidence for the molecular imprinting phenomenon. The linear relationship between the frequency shifts and the concentration of analyte in the range of 10-200 microM was obtained. LPD proves to be a powerful method for imprinting titanium oxide thin films. PMID- 15801450 TI - Lysozyme adsorption studies at the silica/water interface using dual polarization interferometry. AB - Lysozyme adsorption at the silica/water interface has been studied using a new analytical technique called dual polarization interferometry. This laboratory based technique allows the build up or removal of molecular layers adsorbing or reacting on a lightly doped silicon dioxide (silica) surface to be measured in terms of thickness and refractive index changes with time. Lysozyme adsorption was studied at a range of concentrations from 0.03 to 4.0 g dm(-3) and at both pH 4 and pH 7. Adsorbed layers ranging from 14 to 43 +/- 1 A in thickness and 0.21 to 2.36 +/- 0.05 mg m(-2) in mass coverage were observed at pH 4 with increasing lysozyme concentration, indicating a strong deformation of the monolayer over the low concentration range and the formation of an almost complete sideways-on bilayer toward the high concentration of 4 g dm(-3). At pH 7, the thickness of adsorbed layers varied from 16 to 54 +/- 1 A with significantly higher surface coverage (0.74 to 3.29 +/- 0.05 mg m(-2)), again indicating structural deformation during the initial monolayer formation, followed by a gradual transition to bilayer adsorption over the high concentration end. The pH recycling performed at a fixed lysozyme concentration of 1.0 g dm(-3) indicated a broadly reversible adsorption regardless of whether the pH was cycled from pH 7 to pH 4 and back again or vice versa. These observations are in good agreement with earlier studies undertaken using neutron reflection although the fine details of molecular orientations in the layers differ subtly. PMID- 15801460 TI - Permeability and conductivity of red blood cell templated polyelectrolyte capsules coated with supplementary layers. AB - The permeability of ions and small polar molecules through polyelectrolyte multilayer capsules templated on red blood cells was studied by means of confocal microscopy and electrorotation. Capsules were obtained by removing the cell after polyelectrolyte multilayer formation by means of NaOCl treatment. This procedure results in cross-linking of poly(allylamine hydrochloride) (PAH) molecules and destroying poly(styrene sulfonate) (PSS) within the multilayer. Capsules are obtained being remarkably different from layer-by-layer (LbL) capsules. These capsules are rather permeable for low as well as for high molecular weight species. However, upon adsorption of extra polyelectrolyte layers the permeability decreased remarkably. The assembly of six supplementary layers of PAH and PSS rendered the capsule almost impermeable for fluorescein. Resealing by supplementary layers is a potential means for filling and release control. By means of electrorotation measurements, it was shown that the capsule walls obtained isolating properties in electrolyte solutions. Conclusions are drawn concerning the mechanism of permeability through cell templated polyelectrolyte multilayer capsules. PMID- 15801458 TI - Probing the orientation of surface-immobilized immunoglobulin G by time-of-flight secondary ion mass spectrometry. AB - Static time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a powerful surface analysis technique for the characterization of protein films because of its chemical selectivity and surface sensitivity. In this study, static ToF-SIMS and principal component analysis (PCA), a multivariate data analysis method, were combined to probe the orientation of surface-immobilized immunoglobulin G (IgG). IgG orientation can enhance its ability to detect its antigen in immunoassay techniques. The IgG used in this work is the mouse monoclonal anti-human chorionic gonadotropin (anti-hCG). Anti-hCG films on different well-defined substrates have been studied using its F(ab')2 and Fc fragments as references. Atomic force microscopy was used to characterize these protein films before static ToF-SIMS analysis. The results from PCA of ToF-SIMS spectra were related to the antibody primary amino acid composition and its three-dimensional structure. PMID- 15801461 TI - Electrochemical control of protein monolayers at indium tin oxide surfaces for the reagentless optical biosensing of nitric oxide. AB - Cytochrome c has been immobilized onto functionalized, optically transparent indium tin oxide (ITO) electrodes by covalent and electrostatic techniques. Covalent immobilization was achieved by the formation of a disulfide bond between N-succinimidyl 3-(2-pyridyldithio)propionate-(SPDP-) modified cytochrome c and SPDP-silanized ITO. Additionally, ITO electrodes have been modified with the bifunctional reagent 1,12-dodecanedicarboxylic acid (DDCA), resulting in formation of a carboxylic acid-terminated monolayer. Covalent protein attachment to the DDCA-functionalized ITO was achieved with the cross-linker 1-[3 (dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride. Electrostatic attachment of the protein involved ion-pair and hydrogen-bond interactions between the terminating carboxylic acid groups of the DDCA-functionalized ITO and the primary amine groups of the lysine residues of cytochrome c. The electrostatic interaction between the cytochrome c and the functionalized ITO resulted in greater rotational mobility of the protein at the electrode surface, leading to ca. 63% electroactivity, as compared to ca. 41% electroactivity for the covalently immobilized protein. The redox state of the electrostatically bound cytochrome c monolayers could be electrochemically switched between ferric and ferrous forms. Electrochemical control of the bound protein was used to regenerate the biosensing surface following binding of nitric oxide (NO). Ligation of NO with the cytochrome c was monitored by measurement of the change of absorbance intensity at 416 nm. Through application of a negative potential, the cytochrome c was reduced from the ferric to the ferrous form, which led to the removal of the ligated NO. Application of a positive potential regenerated the ferric cytochrome c, enabling multiple repeat measurements of NO. Such electrochemical control of proteins immobilized on transparent electrodes enables the optical biosensing of analyte targets without recourse to exogenous reagents. PMID- 15801462 TI - Ciprofloxacin-protected gold nanoparticles. AB - The antibacterial drug ciprofloxacin (cfH) has been used to protect gold nanoparticles of two different mean diameters, 4 and 20 nm. The protection is complete with about 65 and 585 cfH molecules covering 4 and 15 nm particles, respectively. The nature of binding has been investigated by several analytical techniques. The nitrogen atom of the NH moiety of piperazine group binds on the gold surface, as revealed by voltammetric and spectroscopic studies. The cfH adsorbed particles are stable in the dry state as well as at room temperature, and as a result, redispersion is possible. The rate of release of the drug molecule from the nanoparticles is more in the basic medium than in pure water, and the kinetics depend on the size of the particle; faster desorption is seen in smaller particles. The bound cfH is fluorescent, and this property could be used in biological investigations. This study shows that metal nanoparticles could be useful carriers for cfH and fluoroquinolone molecules. Most of the bound molecules could be released over an extended period of time. PMID- 15801470 TI - Photochemical assembly of gold nanoparticles utilizing the photodimerization of thymine. AB - Irradiation of UV light to the solution of gold nanoparticles modified with thymine units resulted in the formation of aggregates comprising chemical cross linking gold nanoparticles through the photodimerization of the thymine units. Transmission electron microscopy and UV-visible absorption measurement showed the aggregates consisting of the gold nanoparticles. The effect of thymine unit density on the nanoparticle surface and the concentration of the gold nanoparticles in solution to the aggregation process were studied by UV-visible absorption measurement. PMID- 15801478 TI - Protein diffusion in charged nanotubes: "on-off"' behavior of molecular transport. PMID- 15801482 TI - Dietary intake of Cruciferous vegetables, Glutathione S-transferase (GST) polymorphisms and lung cancer risk in a Caucasian population. AB - OBJECTIVE: To evaluate possible interactions between dietary intake of cruciferous vegetables and the glutathione s-transferase mu and theta (GSTM1 and GSTT1) genotypes in lung cancer risk. METHODS: Hospital-based case-control study of 716 Caucasian lung cancer cases and 939 spouse and friend controls conducted in Boston, Massachussetts between 1992 and 2000. Dietary intake was collected through a food frequency questionnaire and blood was obtained for genotyping. Logistic regression models were adjusted for age, gender, total calories and smoking variables. RESULTS: Higher intakes of cruciferous vegetables reduced lung cancer risk among GSTM1 present individuals (odds ratio (OR)highest versus lowest tertile = 0.61, 95% confidence interval (CI) = 0.39-0.95) but not among GSTM1 null individuals (OR(highest versus lowest tertile) = 1.15, 95% CI = 0.78-1.68). We observed statistically significant interactions between GSTM1 and cruciferous vegetable intake overall (likelihood ratio test (LRT): p = 0.05) and among current smokers (LRT: p = 0.01). No significant interactions were observed for GSTT1 or the combined GSTM1/T1 genotype. CONCLUSIONS: In our study, higher cruciferous vegetable intake reduced lung cancer risk only among individuals with the GSTM1 present genotype. Our findings differed from prior studies that specifically assessed isothiocyanates found in cruciferous vegetables or evaluated Asian study populations with higher levels of cruciferous vegetable consumption. PMID- 15801483 TI - Trends in breast cancer mortality rates by region of the United States, 1950 1999. AB - INTRODUCTION: Limited information is available on trends in breast cancer mortality by region of the country. METHODS: Rates for broad age groups were calculated from 1950 to 1999 for whites and 1970-1999 for blacks for four census regions and 508 state economic areas of the United States. RESULTS: For white women ages 50-64 years, the mortality relative risk [RR] for the Northeast compared to the South was 1.48 in 1950-1959 and 1.15 in 1990-1999. Rates increased in all regions from the 1950s to 1960s but more substantially in the South, increased slightly in the 1970s in all regions, declined slightly in the Northeast, Midwest and West but not in the South in the 1980s, and declined more in the Northeast, Midwest and West than in the South in the 1990s. Among similarly aged black women, the RRs for the Northeast compared to the South were 1.13 and 1.0 in 1970-1979 and 1990-1999, respectively. Among these women, rates increased in all regions in the 1980s; in the 1990s rates declined in the Northeast, Midwest and West but continued to increase in the South. CONCLUSION: The historically lower breast cancer mortality rates in the South have been eroded because of relatively less favorable trends in the South. PMID- 15801484 TI - Maternal smoking during pregnancy and risk of brain tumors in the offspring. A prospective study of 1.4 million Swedish births. AB - OBJECTIVE: Studies of the effect of maternal smoking during pregnancy on development of brain tumors in the offspring generally have found no increase in risk but most have mainly relied on retrospective exposure assessment. We conducted a prospective study on a large birth cohort in Sweden. METHODS: Women giving birth during 1983-1997 were classified as smokers or non-smokers based on information ascertained at the first prenatal visit and recorded in the Swedish Birth Register. Follow-up of brain tumor incidence among offspring through 1997 was achieved by linkage with the Swedish Cancer Register. Hazard ratios were estimated using Cox proportional hazard regression, adjusting for demographic characteristics available in the Birth Register. RESULTS: Brain tumors (n=480) occurred at a rate of 4.5 cases per 100,000 person-years. Children of women who smoked during pregnancy had an increased incidence of brain tumors (hazard ratio = 1.24; 95% confidence interval: 1.01-1.53). The increase in risk was similar for benign and malignant tumors, and was most apparent for astrocytoma. The effect of smoking on the occurrence of brain tumors was seen most strongly among 2-4 year old children. CONCLUSIONS: These results support a role for maternal smoking during pregnancy in the etiology of childhood brain tumors. Our findings should be confirmed in other prospective studies. PMID- 15801485 TI - A case-control study of occupational risk factors for bladder cancer in Canada. AB - OBJECTIVE: To investigate occupational risk factors for bladder cancer in seven Canadian provinces. METHODS: We analysed a population-based case-control dataset of 887 individuals with incident, histologically confirmed bladder cancer between 1994 and 1997. Controls (2847) frequency matched for age and gender were surveyed in 1996. Questionnaires were returned by about 60% of subjects. Odds ratios (ORs) for occupations and self-reported exposures were adjusted for province, age, race, smoking, and several dietary factors, using unconditional logistic regression. RESULTS: Statistically significant increased risks were observed among men employed as hairdressers (OR = 3.42; 1.09-10.8), primary metal workers (OR = 2.40; 1.29-4.50), miners (OR = 1.94; 1.18-3.17), and automechanics (OR = 1.69; 1.02-2.82). Primary metal workers and automechanics showed evidence of an employment duration-response trend. Modest elevated risks that were not significant were also observed for male government inspectors, printers, firefighters, general labourers, and welders. A duration-response trend was evident for government inspectors and general labourers. For females, significant elevations were observed among lumber processors (OR = 8.78; 1.28-60.1), general labourers (OR = 2.18; 1.05-4.52), nurses (OR = 1.54; 1.03-2.31), and general clerks (OR = 1.48; 1.01-2.17). The latter showed a positive duration-response trend. CONCLUSIONS: This study found a statistically significant excess risk of bladder cancer, with a duration-response trend, among male primary metal workers and automechanics, and female office workers engaged in general clerical duties. PMID- 15801486 TI - The incidence of cancer in people with intellectual disabilities. AB - OBJECTIVE: During the last 50 years there have been significant improvements in life expectancy among people with intellectual disability (ID), and so their incidence of age-associated diseases, such as cancer, is rising. The aim of this study was to compare the rate of cancer in people with ID with that found in the general population. METHODS: Information on 9409 individuals registered with the Disability Services Commission of Western Australia was linked to the State Cancer Registry, with 200 cases of cancer detected over 156,729 person-years. Standardised incidence ratios (SIRs) and 95% confidence intervals were calculated for both sexes separately by 5-year age groups for the period 1982-2001. The same procedures were adopted in the estimation of SIRs for specific types of cancers. RESULTS: The age-standardised incidence of all cancers in people with ID was not significantly different from the general population. However, males with ID were observed to have a significantly increased risk of leukaemia, brain and stomach cancers, and a reduced risk of prostate cancer, while leukaemia, corpus uteri and colorectal cancers were significantly higher in females. CONCLUSIONS: Health practitioners need to be aware that with improvements in life expectancy the incidence of cancer in people with ID is likely to rise. More proactive health promotion campaigns may be needed for people with ID, who are likely to be poor users of screening services and whose symptoms may not be reported until they are in more advanced, less treatable stages of disease. PMID- 15801487 TI - How representative are the surveillance, epidemiology, and end results (SEER) program cancer data of the United States? AB - OBJECTIVE: Data from the 11 Surveillance, Epidemiology, and End Results tumor registries cover 14% of the U.S. population and are often used to provide national estimates of cancer incidence and survival. Cancer mortality data for the U.S. and for SEER are compared to assess the representation of SEER to the U.S. METHODS: Comparisons between US and SEER cancer mortality were made for 16 of the leading causes of cancer death. Cancer site-specific comparisons were also made by race and sex. In addition, tobacco-related cancers were considered. Analyses were performed for the years when all 11 SEER registries were included in the SEER program. Poisson regression was used to estimate site-specific cancer mortality rate ratios between the U.S. and SEER. RESULTS: Cancer site-specific mortality rates derived from SEER data tend to under-represent the U.S. cancer mortality experience for white males and females and black males. Under representation is observed across the majority of SEER registries, with the highest amount of under representation in Utah and New Mexico. Under representation of SEER data compared with US data is noticeably greater among tobacco-related cancers, particularly in Utah and New Mexico. CONCLUSION: For certain cancer sites, particularly tobacco-related cancers, the SEER coverage population is not representative of the U.S. population. PMID- 15801488 TI - Exercise interventions for cancer patients: systematic review of controlled trials. AB - OBJECTIVE: To systematically review controlled trials investigating the effects of exercise interventions in cancer patients. METHODS: Studies were located through searching seven electronic databases (Medline, Embase, Cochrane Library, CancerLit, PsycInfo, Cinahl, SportDiscus), scanning reference lists of relevant articles, contacting experts (n = 20), and checking the contents lists of journals available through ZETOC (Electronic Table of Contents). To be included, trials had to be prospective, controlled, involve participants diagnosed with cancer and test an exercise intervention. Types of outcome were not restricted. Two reviewers independently applied the selection criteria. RESULTS: Thirty-three controlled trials (including 25 randomized trials) were included in the review. There was some evidence that physical function was increased among those who exercised. Furthermore, symptoms of fatigue did not appear to be increased and there were few adverse effects reported. There was insufficient evidence to determine effects on other outcomes, such as quality of life, with results hampered by the heterogeneity between studies as well as poor methodological quality. Data were also lacking on the long term effects of exercise relating to cancer recurrence or survival. CONCLUSIONS: There is preliminary evidence that exercise interventions for cancer patients can lead to moderate increases in physical function and are not associated with increased symptoms of fatigue. However, it is impossible from current evidence to determine whether exercise has long term beneficial effects on survival or quality of life. PMID- 15801489 TI - Rationale and design of the National Program of Cancer Registries' Breast, Colon, and Prostate Cancer Patterns of Care Study. AB - BACKGROUND: Investigators from the Centers for Disease Control and Prevention (CDC), National Program of Cancer Registries (NPCR), are collaborating with public health professionals from seven states and the District of Columbia to conduct the Patterns of Care study to assess the quality of cancer data and to determine whether stage-specific treatments are being carried out. METHODS: To assess the quality and completeness of cancer care data in the United States, trained staff from the Patterns of Care study are abstracting medical records to obtain detailed clinical data on treatment, tumor characteristics, stage at diagnosis, and demographics of representative samples of patients diagnosed with breast, colon, and prostate cancer. Altogether staff from each of the eight participating cancer registries will abstract 500 cases of breast, prostate, and colon/rectum/anus cancer for the CONCORD study and an additional 150 cases of localized breast cancer, 100 cases of stage III colon cancer, and 100 cases of localized prostate cancer for the Patterns of Care study. Chi-square tests will be used to compare routine registry data with re-abstracted data. The investigators will use logistic regression techniques to describe the characteristics of patients with localized breast and prostate cancer and stage III colon cancer. Age, race, sex, type of insurance, and comorbidity will be examined as predictors of the use of those treatments that are consistent with consensus guidelines. The investigators plan to use data from the CONCORD study to determine whether treatment factors are the reason for the reported differences between relative survival rates in the United States and Europe. CONCLUSIONS: Results from the methodology used in the Patterns of Care study will provide, for the first time, detailed information about the quality and completeness of stage and treatment data that are routinely collected by states participating in the NPCR. It will add significantly to our understanding of factors that determine receipt of treatment in compliance with established guidelines. As part of the CONCORD study, it will also examine differences in survival among cancer patients with breast, prostate, and colon/rectum/anus cancers in the United States and Europe. PMID- 15801490 TI - Alcohol intake and risk of non-Hodgkin lymphoma in men and women. AB - OBJECTIVE: The effect of alcohol intake on risk of NHL is unclear. We therefore conducted a population-based case-control study to examine the association between alcohol and NHL risk. METHODS: 613 NHL cases and 480 population controls in Sweden reported their average consumption of beer, wine, and liquor 2 years before the study. Unconditional logistic regression was used to estimate the odds ratios (OR) and corresponding 95% confidence intervals (CI) for associations between alcohol intake and NHL risk. RESULTS: Intake of total alcohol, beer, wine, or liquor was not associated with risk of overall NHL. There was no difference in risk of NHL among those who habitually consumed above 19.1 g of ethanol per day, compared to those who consumed on average 0-2.2 g of ethanol per day (OR = 1.2 (95% CI: 0.8, 1.7); Ptrend = 0.29). However, the association was significantly positive among males (OR = 1.8 (95% CI: 1.1, 2.9); Ptrend = 0.06). Total alcohol, beer, wine, or liquor intake was not associated with any major histopathologic subtype of NHL examined, apart from an association between high wine consumption and increased risk of chronic lymphocytic leukemia. CONCLUSIONS: Alcohol does not appear to be a major etiologic factor for overall NHL, nor its common subtypes. PMID- 15801491 TI - Polymorphic variation of Cyp1A1 is associated with the risk of gastric cardia cancer: a prospective case-cohort study of cytochrome P-450 1A1 and GST enzymes. AB - OBJECTIVE: To determine if genetic polymorphisms of CYP1A1, GSTM1, GSTP1, or GSTT1 are associated with an increased risk of developing esophageal squamous cell carcinoma (ESCC), gastric cardia cancer (GCC), or either in a high-risk Asian population. METHODS: We conducted a case-cohort analysis with 5 years of prospective follow-up. The analytical cohort contained 642 individuals who participated in either the Dysplasia Trial (DT) or the General Population (GPT) of the Nutrition Intervention Trials conducted in Linxian, China, and included 131 cases of ESCC and 90 cases of GCC. Genotyping analysis was performed on DNA extracted from red blood cells using a PureGene kit (Gentra Systems, Inc., Minneapolis, MN) and real-time PCR analysis amplification (Taq-Man). Relative risks and 95% confidence intervals were estimated using the case - cohort estimator for the Cox proportional hazards models. p-values from nested models with genotyping variables came from score tests. RESULTS: The relative risks for developing ESCC, GCC, or either cancer were calculated in the entire analytic cohort for GSTM1, P1*B (A313G), and T1 and CYP1A1*2A (T3801C) and *2C (A2455G) genotypes, and no significant associations were identified. However, because of the difference in cancer risks between the DT (9.3 cases per 1000 person years) and the GPT (5.3 cases), the analytical cohort was stratified by trial; the DT participants who were heterozygous or homozygous for the variant-allele at CYP1A1*2A had a reduced risk for developing GCC (adjusted RR (95% CI) 0.47 (0.23 1.00) p = 0.037). CONCLUSIONS: This study found an association for the CYP1A1*2A variant allele and a reduced risk of GCC in people at high risk for development of this disease. This finding is consistent with previous studies suggesting that substrates for the cytochrome P-450 1A1 metabolic pathway, such as polycyclic aromatic hydrocarbons, may be etiologically significant in this high-risk region. PMID- 15801492 TI - Association of folate and alcohol with risk of ovarian cancer in a prospective study of postmenopausal women. AB - Studies evaluating the association of ovarian cancer with alcohol intake are inconsistent, and few have evaluated this association in the context of folate consumption. Dietary folate and alcohol intakes and lifestyle and medical information were collected with self-administered questionnaires in 1986 from postmenopausal women aged 55-69 followed prospectively for 15 years for risk of epithelial ovarian cancer in the Iowa Women's Health Study. Among 27,205 eligible women free of baseline cancer, 147 incident epithelial ovarian cancer cases were identified by linkage to a cancer registry. Compared to the lowest quartile of total folate (food plus supplement) intake, the multivariable risk ratios (RR) for increasing quartiles were 1.0 (referent), 1.59, 1.24, 1.73 (95% confidence interval [CI], 0.90-3.33; p for trend, 0.20). Compared to non-drinkers, the RRs for increasing alcohol intake were 1.0 (referent), 0.78 for 0.01-3.9 g/d, 0.75 for 4.0-9.9 g/d and 0.58 for > or =10 g/d (95% CI, 0.30-1.11; p for trend, 0.08). Among women with alcohol intake > or =4 g/d compared to < 4 g/d, the apparent risk reduction was limited to those with total folate intake > or =331 microg/d (RR, 0.52; 95% CI, 0.22-1.19; p for interaction, 0.04) although this estimate was based on only seven cases. The association did not change appreciably when we excluded tumors of mucinous histology. These findings suggest that alcohol consumption is inversely related to postmenopausal ovarian cancer, and that the association of folate with ovarian cancer may vary by the amount of alcohol consumed. PMID- 15801493 TI - Prospective study of plasma enterolactone and prostate cancer risk (Sweden). AB - OBJECTIVES: Enterolactone, a phytoestrogen produced by the intestinal microflora from precursors in plant foods, has been postulated to protect against hormone dependent cancers. We studied the association between plasma enterolactone and risk of prostate cancer. METHODS: In the Northern Sweden Health and Disease Cohort, enterolactone concentrations were measured by time-resolved fluoroimmunoassay in plasma taken from 265 men who were diagnosed with prostate cancer at a mean time of 5 years after blood collection, and in plasma from 525 control men, matched for age and date of blood collection. RESULTS: There was no significant association between quartiles of plasma enterolactone and risk of prostate cancer. Odds ratios for prostate cancer, estimated by conditional logistic regression for increasing concentrations of enterolactone in quartiles were 1.00 (referent), 0.81 (95% confidence interval 0.52-1.27), 1.03 (0.67-1.58), and 1.22 (0.80-1.86). Adjustments for body mass index (BMI), smoking status and stratification for age, lag time, storage time and tumour characteristics did not materially alter risk estimates. Men with very low enterolactone levels, however, had significantly higher risk of prostate cancer, odds ratio for bottom decile versus all other deciles was 1.68 (1.03-2.74). CONCLUSIONS: Our results do not support the hypothesis that enterolactone formed from dietary lignans protects against prostate cancer. PMID- 15801494 TI - Damage to the human quadriceps muscle from eccentric exercise and the training effect. AB - Nine participants performed two bouts of a step exercise, during which the quadriceps muscle of one leg acted eccentrically. Before and after the exercise, isokinetic torque was measured over a range of knee angles to determine the optimum angle for torque. Immediately after the first bout of exercise, the quadriceps showed a significant (P < 0.05) shift of 15.6 +/- 1.4 degrees (mean +/ sx) of its optimum angle in the direction of longer lengths, suggesting the presence of damage. A drop in peak torque, together with delayed soreness and swelling, confirmed that damage to muscle fibres had occurred. After the second bout of exercise, 8 days later, the shift in optimum angle was 10.4 +/- 1.0 degrees, which was significantly less than after the first bout (P < 0.05). Other indicators of damage were also reduced. In addition, the muscle exhibited a sustained shift in optimum angle (3.4 +/- 0.9 degrees), suggesting that some adaptation had taken place after the first bout of exercise. We conclude that muscles like the quadriceps can show evidence of damage after a specific programme of eccentric exercise, followed by an adaptation response. This is despite the fact that the quadriceps routinely undergoes eccentric contractions in everyday activities. PMID- 15801495 TI - The effect of single and repeated bouts of prolonged cycling and circadian variation on saliva flow rate, immunoglobulin A and alpha-amylase responses. AB - The purpose of this study was to establish the effect of exercise at different times of day on saliva flow rate, immunoglobulin A (sIgA) concentration and secretion rate, and alpha-amylase activity, and to establish how these parameters change following a second exercise bout performed on the same day. In a counterbalanced design, eight male volunteers participated in three experimental trials separated by at least 4 days. On the trial with afternoon exercise only, the participants cycled for 2 h at 60% VO2max starting at 14:00 h. On the other two trials, participants performed either two bouts of exercise at 60% VO2max for 2 h (the first started at 09:00 h and the second started at 14:00 h) or a separate resting trial. Unstimulated saliva samples were obtained 10 min before exercise, after 58 - 60 min and during the last 2 min of exercise, and at 1 h and 2 h after exercise. Venous blood samples were taken 5 min before exercise and immediately after exercise for both bouts. Participants remained fasted between 23:00 h on the day before the trials and 18:00 h on the day of the trial. Circadian variations were found in sIgA concentration, which decreased with time from its highest value in the early morning to its lowest value in the evening, and salivary alpha-amylase secretion rate, which increased from its lowest value in the morning to its highest value in the late afternoon. Cycling at 60% VO2max for 2 h significantly decreased saliva flow rate, increased sIgA concentration and alpha-amylase activity, but did not influence sIgA secretion rate. Performing prolonged cycling at different times of day did not differentially affect the salivary and plasma hormonal responses in the short term. Performance of a second prolonged exercise bout elicited a greater plasma stress hormone response but did not appear to compromise oral immunity acutely. These findings also suggest that, in terms of saliva secretion, sIgA and alpha-amylase responses, a 3 h rest is enough to recover from previous strenuous exercise. During such exercise, sympathetic stimulation appears to be strong enough to inhibit saliva flow rate; however, it appears that it does not increase sIgA output via transcytosis. PMID- 15801496 TI - Training of elite cyclists: effects on mood state and selected hormonal responses. AB - The psychobiological status of cyclists over a period of 8 months of training was assessed by measuring the sympatho-adrenal level, the central noradrenergic activity, and the cortisol-testosterone ratio status noninvasively. Alteration of these indices after a large increase in training load lasting 4 days (+ 187%) was also examined. Urinary excretion of methoxyamines (metanephrine, normetanephrine) and 3-methoxy-4-hydroxyphenylglycol sulphate (MHPG-S), and salivary concentrations of cortisol and testosterone, were measured in 12 national cyclists after a non-specific training period and 48 h before 4 days of increased training (T1), after these 4 days (T2) and at the end of 4 months of specific training (T3). Urinary and salivary samples were also collected during a rest period (T0). At each of these times, mood states and ratings of perceived muscle soreness were assessed, and a questionnaire of early clinical symptoms of the overtraining syndrome (Profile of Mood States) was administered. A significant increase in normetanephrine (P < 0.05) and a decrease in the testosterone cortisol ratio (P < 0.05) were observed at T2, while MHPG-S excretion remained unchanged. Over the same periods, increased training loads did not induce mood disturbances. Eight months of training were associated with significant alterations in metanephrine (P < 0.05) and MHPG-S (P < 0.05). These results suggest a dissociation between the neural and endocrine catecholaminergic components systems. Opposite responses between MHPG-S and Profile of Mood States scores show that further investigations are needed to understand the relationship between central noradrenergic function, which is recognized as a regulatory factor of mood, and psychological tests measuring mood. PMID- 15801497 TI - Effects of modified-implement training on fast bowling in cricket. AB - The effects of training with overweight and underweight cricket balls on fast bowling speed and accuracy were investigated in senior club cricket bowlers randomly assigned to either a traditional (n = 9) or modified-implement training (n = 7) group. Both groups performed bowling training three times a week for 10 weeks. The traditional training group bowled only regulation cricket balls (156 g), whereas the modified-implement training group bowled a combination of overweight (161-181 g), underweight (151-131 g) and regulation cricket balls. A radar gun measured the speed of 18 consecutive deliveries for each bowler before, during and after the training period. Video recordings of the deliveries were also analysed to determine bowling accuracy in terms of first-bounce distance from the stumps. Bowling speed, which was initially 108 +/- 5 km h(-1) (mean +/- standard deviation), increased in the modified-implement training group by 4.0 km x h(-1) and in the traditional training group by 1.3 km x h(-1) (difference, 2.7 km x h(-1); 90% confidence limits, 1.2 to 4.2 km x h(-1)). For a minimum worthwhile change of 5 km x h(-1), the chances that the true effect on bowling speed was practically beneficial/trivial/harmful were 1.0/99/< 0.1%. For bowling accuracy, the chances were 1/48/51%. This modified-implement training programme is not a useful training strategy for club cricketers. PMID- 15801498 TI - String vibration dampers do not reduce racket frame vibration transfer to the forearm. AB - In this study, we examined the effect of string vibration damping devices on reducing racket frame vibration transfer to the forearm. Twenty participants volunteered to hold a tennis racket stationary in a forehand and backhand stroking position while tennis balls were fired at 20 m x s(-1) towards two impact locations, the node of vibration and the dead spot. A three-way analysis of variance with repeated measures on damping condition, impact location and stroke condition was performed on the data. The resonant frequency of the hand held racket was found to be approximately 120 Hz. No significant differences in amplitude of vibration at the resonant frequency were found for the wrist or the elbow when damped and non-damped impacts were compared. Impacts at the dead spot produced greater amplitudes of vibration (P < 0.01) but no interaction between impact location and string dampers was evident. The string dampers had no effect on the grip force used or the muscle electrical activity in the forearm after impact. In conclusion, we found that string dampers do not reduce the amount of racket frame vibration received at the forearm. We suggest that string dampers remain a popular accessory among tennis players because of their acoustic effects and psychological support rather than any mechanical advantage. PMID- 15801500 TI - Skeletal muscle glycogen concentration and metabolic responses following a high glycaemic carbohydrate breakfast. AB - The purpose of this study was to examine the influence of a carbohydrate-rich meal on post-prandial metabolic responses and skeletal muscle glycogen concentration. After an overnight fast, eight male recreational/club endurance runners ingested a carbohydrate (CHO) meal (2.5 g CHO x kg(-1) body mass) and biopsies were obtained from the vastus lateralis muscle before and 3 h after the meal. Ingestion of the meal resulted in a 10.6 +/- 2.5% (P < 0.05) increase in muscle glycogen concentration (pre-meal vs post-meal: 314.0 +/- 33.9 vs 347.3 +/- 31.3 mmol x kg(-1) dry weight). Three hours after ingestion, mean serum insulin concentrations had not returned to pre-feeding values (0 min vs 180 min: 45 +/- 4 vs 143 +/- 21 pmol x l(-1)). On a separate occasion, six similar individuals ingested the meal or fasted for a further 3 h during which time expired air samples were collected to estimate the amount of carbohydrate oxidized over the 3 h post-prandial period. It was estimated that about 20% of the carbohydrate consumed was converted into muscle glycogen, and about 12 % was oxidized. We conclude that a meal providing 2.5 g CHO x kg(-1) body mass can increase muscle glycogen stores 3 h after ingestion. However, an estimated 67% of the carbohydrate ingested was unaccounted for and this may have been stored as liver glycogen and/or still be in the gastrointestinal tract. PMID- 15801499 TI - Relation between multidimensional performance characteristics and level of performance in talented youth field hockey players. AB - To determine the relationship between multidimensional performance characteristics and level of performance in talented youth field hockey players, elite youth players (n = 38, mean age 13.2 years, s = 1.26) were compared with sub-elite youth players (n = 88, mean age 14.2 years, s = 1.26) on anthropometric, physiological, technical, tactical and psychological characteristics. Multivariate analyses with performance level and gender as factors, and age as the covariate, showed that the elite youth players scored better than the sub-elite youth players on technical (dribble performance in a peak and repeated shuttle run), tactical (general tactics; tactics for possession and non-possession of the ball) and psychological variables (motivation) (P < 0.05). The most discriminating variables were tactics for possession of the ball, motivation and performance in a slalom dribble. Age discriminated between the two groups, indicating that the elite youth players were younger than the sub-elite players. In the guidance of young talented players to the top as well as in the detection of talented players, more attention has to be paid to tactical qualities, motivation and specific technical skills. PMID- 15801501 TI - The effects of wind and altitude in the 400-m sprint. AB - In this paper I use a mathematical model to simulate the effect of wind and altitude on men's and women's 4400-m race performances. Both wind speed and direction were altered to calculate the effect on the velocity profile and the final time of the sprinter. The simulation shows that for a constant wind velocity, changing the wind direction can produce a large variation in the race time and velocity profile. A wind of velocity 2 m x s(-1) is generally a disadvantage to the 400-m runner but this is not so for all wind directions. Constant winds blowing from some directions can provide favourable conditions for the one-lap runner. Differences between the running lanes can be reduced or exaggerated depending on the wind direction. For example, a wind blowing behind the runner in the back straight increases the advantage of lane 8 over lane 1. Wind conditions can change the velocity profile and in some circumstances produce a maximum velocity much later than is evident in windless conditions. Lower air density at altitude produces a time advantage of around 0.06 s for men (0.07 s for women) for each 500-m increase in elevation. PMID- 15801502 TI - Thermoregulatory responses to exercise: relative versus absolute intensity. AB - The purpose of the present study was to re-examine the relationship between deep body temperature and relative exercise intensity, during running rather than cycling (Saltin and Hermansen, 1966). Twenty male competitive and recreational distance runners, aged 22 + 0.9 years (mean +/- sx), were selected to form two groups, one with high maximal oxygen uptake (VO2max) values (72.8 +/- 0.8 ml x kg(-1) x min(-1)) and the other with moderate values (59.4 +/- 0.7 ml x kg(-1) x min(-1)). The participants completed two 60 min constant-paced treadmill runs at a common speed (absolute intensity) of 10.5 km x h(-1) and at a relative exercise intensity at a speed equivalent to 65% of VO2max. During the relative exercise intensity trial, no differences were found in rectal temperature, skin temperature or heart rate between groups. However, when running at the common speed, differences were identified in rectal temperature. At 60 min, rectal temperature was 37.70 +/- 0.19 degrees C and 38.19 +/- 0.11 degrees C for the high and moderate VO2max groups, respectively (P < 0.05). Sweat lost was significantly higher in the moderate VO2max group (moderate: 1.05 +/- 0.06 kg x h(-1); high: 0.82 +/- 0.08 kg x h(-1); P < 0.05). Heart rates were also different between groups over the first 20 min during the common speed trial (P < 0.05). The results of the present study support the findings of Saltin and Hermansen (1966), in that the set-point at which temperature is maintained is related to the relative exercise intensity. PMID- 15801503 TI - No effect of fluid intake on neutrophil responses to prolonged cycling. AB - Ingesting carbohydrate beverages during prolonged exercise is associated with fewer numbers of circulating neutrophils and attenuated neutrophil functional responses, yet there is little information about the effect of fluid intake alone on immune responses to prolonged exercise. The aim of this study was to examine the influence of regular fluid ingestion compared with no fluid ingestion on plasma cortisol, circulating neutrophil and lipopolysaccharide (LPS)-stimulated neutrophil degranulation responses to prolonged cycling. In a randomized design, nine recreationally active males cycled for 2 h at 65% VO2max on two occasions with either fluid ingestion (lemon-flavoured water, fluid trial) before and during the exercise, or with no fluid intake at all (no fluid trial). Venous blood samples were obtained at rest, immediately after exercise and 1 h after exercise. Immediately after exercise, the plasma cortisol concentration was significantly higher in the no fluid trial than in the fluid trial (592 +/- 62 vs 670 +/- 63 nmol x l(-1), P < 0.05). Circulating numbers of neutrophils increased 4.5-fold (P < 0.01) and LPS-stimulated elastase release per neutrophil decreased 34 +/- 7% (P < 0.01) immediately after exercise; there were no differences between trials. These results suggest that in ambient environmental conditions, fluid ingestion alone has a negligible effect on circulating neutrophil and LPS stimulated neutrophil degranulation responses to prolonged exercise. PMID- 15801504 TI - Semantic deficits in children with language impairments: issues for clinical assessment. AB - Children with language impairments demonstrate a broad range of semantic difficulties, including problems with new word acquisition, storage and organization of known words, and lexical access/ retrieval. Unfortunately, assessments of children's semantic skills are often limited to measures of receptive and expressive vocabulary size. As a result, the semantic deficits of these children may not receive the attention they need. This article explores the word-learning, lexical storage, and lexical access skills of children with language impairments and the theories that account for their performance. Our review culminates with specific recommendations for speech-language pathologists to improve the breadth of their semantic assessments. PMID- 15801505 TI - Learning new words from storybooks: an efficacy study with at-risk kindergartners. AB - PURPOSE: The extant literature suggests that exposure to novel vocabulary words through repeated readings of storybooks influences children's word learning, and that adult elaboration of words in context can accelerate vocabulary growth. This study examined the influence of small-group storybook reading sessions on the acquisition of vocabulary words for at-risk kindergartners, and the impact of word elaboration on learning. An additional goal was to study differential responses to treatment for children with high versus low vocabulary skill. METHOD: Using a pretest-posttest comparison group research design, 57 kindergartners were randomly assigned to a treatment (n = 29) or comparison (n = 28) group. Children were also differentiated into high (n = 31) versus low (n = 26) vocabulary skill groups using scores on a standardized receptive vocabulary test. Children in the treatment group completed 20 small-group storybook reading sessions during which they were exposed to 60 novel words randomly assigned to non-elaborated and elaborated conditions. Pre- and posttest examined the quality of children's definitions for the 60 novel words. RESULTS: Overall, word-learning gains were modest. Children in the treatment group made significantly greater gains in elaborated words relative to children in the comparison group; no influence of storybook reading exposure was seen for non-elaborated words. Children with low vocabulary scores made the greatest gains on elaborated words. CLINICAL IMPLICATIONS: Suggestions are offered for using storybooks as a clinical tool for fostering vocabulary development. As an efficacy study, results should inform future applied research on word learning for at-risk children. PMID- 15801506 TI - Use of a parent questionnaire in emergent and early literacy assessment of preschool children. AB - PURPOSE: The purpose of this project was to evaluate the use of a parent questionnaire in the assessment of emergent and early literacy skills of preschool children with language impairments (LI) and their typically developing (TD) peers to determine the relationship between parent report and more formal measures of early literacy, as well as to compare home literacy practices of these two groups of children. METHOD: Preschool children with LI (n = 17) were referred by area school district speech-language pathologists, and age-matched peers (n = 20) were recruited from local preschools. Examiner-administered measures of early literacy included assessment of rhyme, knowledge of environmental print, knowledge of print conventions, and letter-name and letter sound knowledge, as well as analyses of narrative abilities. The parent questionnaire included 31 closed questions with responses chosen on a 5-point scale that addressed a number of domains of knowledge: phonological awareness, response to print in the environment, alphabet knowledge, interactions around books, writing, orientation to literacy, and parent practices, in addition to a number of open-ended questions. RESULTS: Results revealed a strong relationship between examiner-administered measures of early literacy and parent report for the children with LI only. Parents of children with LI rated their children more poorly across domains of literacy knowledge than did parents of the comparison group. Finally, a comparison of home literacy practices found differences between the groups in time spent watching television, age parents began reading to the child, and number of books owned by the child. CLINICAL IMPLICATIONS: Findings support the use of parent report as a tool in the assessment of emergent and early literacy knowledge and skills for children with LI. PMID- 15801507 TI - Serving English language learners in public school settings: a national survey. AB - PURPOSE: In 1990, 1,145 public school speech-language pathologists (SLPs) across the United States were surveyed regarding service delivery to English language learner (ELL) students (C. A. Roseberry-McKibbin & G. E. Eicholtz, 1994). In 2001, the survey was replicated with a larger national sample (N = 1,736). The first purpose of the current study was to conduct an in-depth analysis of the 2001 survey results in terms of relationships between variables in respondents' backgrounds (e.g., region of the United States and coursework in service delivery to ELL students) and perceived problems in providing service delivery to ELL students. The second purpose of the current study was to compare answers from the 1990 and 2001 surveys in terms of similarities and differences. METHOD: Six thousand surveys were mailed out to a randomly and independently selected sample of public school SLPs across the United States. One thousand seven hundred thirty six surveys were returned and analyzed. RESULTS: Results indicated both similarities and differences between answers from the 1990 and 2001 surveys. Results from an in-depth analysis of the current survey found that respondents from the West had the most coursework in service delivery to ELL students. Respondents from the West and the Southwest regions of the United States perceived all service delivery problems with ELL students as occurring less frequently than did their counterparts from other regions. Respondents with more university coursework perceived "lack of appropriate less biased assessment instruments" as a more frequently occurring problem than did respondents with less university coursework. CLINICAL IMPLICATIONS: Universities need to offer coursework regarding service delivery to ELL students. A particularly important area that courses should address is less biased assessment of ELL students. Other implications and directions for future research are discussed. PMID- 15801508 TI - The use of two language tests to identify pragmatic language problems in children with autism spectrum disorders. AB - PURPOSE: Pragmatic language disorders (PLDs) are difficult to diagnose in a cost effective manner, and there are few assessment tools that yield quantitative data. This investigation was designed to determine whether two formal assessment tools would differentiate PLDs in children with autism spectrum disorders (ASDs) from controls matched on verbal IQ and language fundamentals. METHOD: Thirty-four matched participants were given the Test of Pragmatic Language (TOPL; D. Phelps Terasaki & T. Phelps-Gunn, 1992) and the Strong Narrative Assessment Procedure (SNAP; C. J. Strong, 1998). RESULTS: Participants with ASDs had significantly poorer scores than controls on the TOPL. On the SNAP, the children with ASDs performed similarly to controls on syntax, cohesion, story grammar, and completeness of episodes. The controls performed significantly better only on the ability to answer inferential questions. CLINICAL IMPLICATIONS: The TOPL was effective in differentiating PLDs in children with ASDs when performance was compared tomatched controls. The SNAP did not clearly differentiatelanguage problems in these two groups. Research is needed to develop formal assessment tools that target the unique language disabilities of high-functioning individuals with ASDs. PMID- 15801509 TI - "Do I know what I need to do?" A social communication intervention for children with complex clinical profiles. AB - PURPOSE: Speech-language pathologists frequently address social communication difficulties in children with diverse clinical profiles. The purpose of this study was to investigate the feasibility of a social communication intervention for a school-age child with a complex cognitive and behavioral profile secondary to diagnosis of a fetal alcohol spectrum disorder. METHOD: A case study is presented to describe the implementation of the intervention targeting mental state verb production and social cognitive skills. The intervention included group role play of social scripts and a checklist to elicit the participant's statements about others' perspectives and strategies for completing the social script. Treatment data monitored the participant's responses to the checklist questions. Probe sessions, consisting of theory of mind false belief tasks, were used to examine mental state verb use. RESULTS: Treatment data demonstrated that the participant stated more strategies in response to checklist questions. The participant did not produce any mental state verbs during baseline probes, but did produce mental state verbs during the treatment phase. CLINICAL IMPLICATIONS: The results support use of this intervention to change children's linguistic and social cognitive skills. Suggestions for extending this intervention to include a generalization plan targeting classroom social communication interactions are provided. PMID- 15801510 TI - The case of Mr B. PMID- 15801511 TI - Incorporation of an invasive object. AB - The author discusses the experience of 'being invaded' that is sometimes communicated by certain severely disturbed patients. The complaint can sometimes be couched in terms of bodily suffering and such patients may state that they have the experience of a 'foreign body' inside. It is suggested that these patients have suffered severe early failure of containment of their projections, while at the same time they have incorporated primitive characteristics of the object that have been powerfully projected into them. An object that invades in this way, it is suggested, experiences a compulsive need to expel unbearable states of mind using others as a repository. The infant incorporates these violent projections as part of his own mental representational system, and normal identification processes are disrupted. There follows impairment of the development of the sense of self. Clinical examples of how the invasive experience manifests itself in the analytic setting and in the transference and countertransference are presented. It is argued that this highly complex form of early subject-object interaction (prior to the differentiation of psyche-soma) is more likely to be found in severely narcissistically disturbed individuals. Some reflections on the origins of invasive phenomena are given. PMID- 15801512 TI - On holding and containing, being and dreaming. AB - Winnicott's concept of holding and Bion's idea of the container-contained are for each of these analysts among his most important contributions to psychoanalytic thought. In this light, it is ironic that the two sets of ideas are so frequently misunderstood and confused with one another. In this paper the author delineates what he believes to be the critical aspects of each of these concepts and illustrates the way in which he uses these ideas in his clinical work. Winnicott's holding is seen as an ontological concept that is primarily concerned with being and its relationship to time. Initially the mother safeguards the infant's continuity of being, in part by insulating him from the 'not-me' aspect of time. Maturation entails the infant's gradually internalizing the mother's holding of the continuity of his being over time and emotional flux. By contrast, Bion's container-contained is centrally concerned with the processing (dreaming) of thoughts derived from lived emotional experience. The idea of the container contained addresses the dynamic interaction of predominantly unconscious thoughts (the contained) and the capacity for dreaming and thinking those thoughts (the container). PMID- 15801513 TI - Between the confusion of tongues and the gift of tongues. Or working as a psychoanalyst in a foreign language. AB - The author worked as a psychoanalyst for 5 years in Germany. In this paper, he attempts to answer the question 'How was it possible that, in spite of his imperfect knowledge of German, notwithstanding a deepening understanding of the language during his residence in the country, he was able to successfully treat so many patients? ' He starts by putting forward some distinctions between the activity of interpretation as translation of the unconscious with the patient in session and the activity of translation of texts. After a brief exegetic review of the myths of Babel and Pentecost, he suggests that the analyst working in a foreign language moves between 'the confusion of tongues' and the 'gift of tongues', that is, between Babel and Pentecost. He presents some vignettes to illustrate typical situations he encountered in his practice. Finally, he draws some conclusions from this experience of psychoanalytic polyglotism, mainly on the basis of the communicative function that modern infant research assigns to affect attunement and verbal language. PMID- 15801514 TI - Making conscious the unconscious in order to modify unconscious processing: some mechanisms of therapeutic change. AB - This paper examines some of the mechanisms through which interpretation aimed primarily at increasing conscious awareness can nonetheless produce unconscious changes, the latter being deemed the basic aim of psychoanalysis. The concept of valency or motivational weight of the interpretation is proposed to assess which forces of the various motivational systems the interpretation mobilizes (hetero/self-preservation, sensual/sexual, attachment, narcissistic, psychobiological regulation etc.), on which of the above-mentioned systems interpretation relies, and which would oppose therapeutic intervention and why. Certain conditions are also analyzed that could explain the so-called 'change through the analytic relationship', pointing out that, despite the major differences between this form of change and change through interpretation, both of them would share certain mechanisms. This conclusion leads to the need to qualify the idea that interpretation would be exclusively aimed at declarative memory, with no effects upon procedural memory. The paper examines the potential consequences for therapeutic techniques derived from recent findings in neuroscience on so-called labile state memory, and proposes the coupling of experiences as one of the analytical instruments used for therapeutic change. A clinical vignette is included to illustrate some of the theoretical and technical aspects considered. PMID- 15801516 TI - Semiotic aspects of the countertransference: some observations on the concepts of the 'immediate object' and the 'interpretant' in the work of Charles S. Peirce. AB - The field of semiotics, established by Charles S. Peirce, is characterised by its recognition of non-linguistic signs and embedment in a communicative interaction; for this reason, it is especially well suited for a semiotic investigation of intersubjective processes. In this paper, the authors show how these intersubjective processes can be understood in semiotic terms within the transference-countertransference setting. Based on a case vignette, the relationship between the 'real object' (e.g. an unconscious fantasy) and the sign (e.g. a particular facial expression) is first demonstrated. In this mediation between sign and referent, an important role is played by the 'immediate object', by which Peirce understood the mental concept of a sign. However, a further component of the Peircian sign is responsible for the emergence of the countertransference, namely, the 'interpretant'. The core of Peircian semiotics, namely the concept of an (infinite) process of signification, sheds light in semiotic terms on the dialectical movement between transference-signs and countertransference-signs, the interpretation and encounter between two subjects. The paper concludes with a discussion of both the interdisciplinary applicability of Peircian semiotics, for example in the context of the neurosciences, and the differences between the Peircian epistemological position and psychoanalytical conceptions of the objective cognition of mental processes. PMID- 15801515 TI - A transitory homosexual passion in the course of an analytic treatment. AB - Do homosexualities express a sexual conflict originating in bisexuality or must we look for their deep determinism in the narcissistic conflict and its vicissitudes? Two clinical sequences from the treatment of a borderline patient, which occurred during years seven and eight of treatment, a period characterised by intense drive turbulence and the unexpected occurrence of a transitory homosexual passion, will enable the reader to follow the psychical movements and evolution of the work of representation. The first sequence throws light on the links between her sudden involvement in this homosexual passion and the traumatic dangers of a breakdown of her ego's capacity for representation, linked to a reactivation in the transference of early and later traumatic traces whose reverberation in the transfero/countertransference dynamic was the source of a painful transference that was responsible for this lateral transference movement. The second sequence shows the links between the strengthening of her capacity for representation, thanks to the working through of the oedipal conflict in its two aspects, and her renunciation of this passion. The roots of this transitory homosexual passion in the deep flaws of her psychical functioning raise metapsychological questions concerning, in particular the dynamic of the 'double' ('twin') and narcissism. PMID- 15801517 TI - Belief, hope and faith. AB - A case of hysteria is presented in order to create a frame of reference for the author's approach to the concepts of hope, belief and faith. A difference between hope as a 'sad passion' (which is here called regressive hope) and hope as a principle of mental functioning is established. The concept of hope will at first always be based on beliefs--either beliefs organised in the paranoid-schizoid position (called here fragmented and delusional beliefs)--or those organised from the depressive position (complex systems of beliefs, which end up being dogmatic); the latter typically occur in neurotics. It is suggested here that there is another possibility for hope, which is based on faith. The meaning of faith is considered here externally to the religious sense. The solid establishment of hope as a principle--based on faith--can be viewed as responsible for the opening up of creative potentials and as one of the main aims of analysis. Such an aim, however requires the establishment of a deep relationship, both in theory and in clinical practice, between the Kleinian question of the depressive position and the Freudian question of the Oedipus complex. PMID- 15801519 TI - What does conceptual research have to offer? PMID- 15801518 TI - Linguistic styles and complementarities in analyzing character. AB - This paper discusses and illustrates some technical implications of D. Liberman's contributions to character analysis. Psychoanalysis is an extended conversation, and character structure is expressed linguistically as part of an interactive linguistic field. Liberman's notions of linguistic styles and communicative interaction define a unique analytic surface from which to examine the syntactic, semantic and pragmatic aspects of character. The presence of a rigid and defensive linguistic style reflects resistance and communicative impasse within the transference and countertransference matrix. A fully interactive communicative experience integrates all aspects of the conversation and allows both participants to take part in a shared semantic field, the precondition to the generation of new meanings. Liberman suggests responding to miscommunication by means of a methodical linguistic attitude. Various such complementary responses are explored through several vignettes, with a particular focus on the interplay of the epic and narrative modes. PMID- 15801520 TI - Language and affects in the analytic practice. PMID- 15801521 TI - On analytic listening. PMID- 15801522 TI - Is free association still at the core of psychoanalysis? PMID- 15801523 TI - Pursuit of the emerging dialogue between psychoanalysis and neuroscience: clinical and research perspectives. PMID- 15801524 TI - Theoretical and technical approaches to the clinical case: advantages and disadvantages of present-day pluralism. PMID- 15801525 TI - Conceptual frontiers: representation and object relations. PMID- 15801526 TI - The plays of Tennessee Williams--a psychoanalytic view. PMID- 15801527 TI - On: What can we learn from psychoanalysis and prospective studies about chemically dependent patients? PMID- 15801528 TI - On: Miss A. PMID- 15801529 TI - [Severe hepatopathies in HIV patients. Epidemiology and diagnosis]. PMID- 15801530 TI - [Severe hepatopathies in HIV patients. Assessment of fibrosis: hepatic biopsy, Fibrotest]. PMID- 15801531 TI - [Severe hepatopathies in patients with HIV-HCV and HIV-HBV coinfection. Management of clinical complications]. PMID- 15801532 TI - [Liver transplantation in patients co-infected with HIV]. PMID- 15801533 TI - [Interpretation of transaminase level increase in patients with HIV-HCV or HIV HBV co-infection]. PMID- 15801534 TI - [Use of antiretroviral agents in cirrhotic patients infected by HIV]. PMID- 15801535 TI - [Interference among antiretroviral, anti-HCV, and immunosuppressive treatments]. PMID- 15801536 TI - [Should HCV patients co-infected with HIV be treated? When? How?]. PMID- 15801537 TI - [Hepatitis B virus resistance]. PMID- 15801538 TI - [Update on the treatment of hepatitis B virus infection in patients infected by the human immunodeficiency virus]. PMID- 15801539 TI - Expression of basolateral and canalicular transporters in rat liver and cultures of primary hepatocytes. AB - Basolateral and canalicular proteins are expressed in the liver and besides their role in the transport of bilirubin, glutathione, hormones and various glucuronides, they also function as transporters of a wide range of drugs. Despite their frequent use in drug research, little is known about the expression of genes coding for transporters in cultures of primary sandwiched hepatocytes. The kinetics of gene expression of canalicular and basolateral membrane transporters in cultures of primary rat sandwiched hepatocytes were investigated, and the expression of cMOAT, spgp, mdrla and mdr2 were shown to be comparable with transcript levels observed in vivo. Strikingly, expression of the basolateral membrane transporter ntcp and oatpl and 2 were dramatically reduced, the level being < 10 and < 5%, respectively, of those found in vivo in rat liver tissue. Notably, mRNA expression of the canalicular membrane transporter mdrlb was increased up to 13-fold. The findings point to a dramatic change in the expression of basolateral and canalicular transporters in cultured hepatocytes, and this should be considered when hepatocytes are used for drug profiling studies. PMID- 15801541 TI - Metabolic properties of the acid and lactone forms of HMG-CoA reductase inhibitors. AB - To gain a better understanding of the metabolic properties between the open acid and lactone form of HMG-CoA reductase inhibitors (statins), the paper focused primarily on characterizing the metabolic properties of statins. We compared the metabolism of the acid and lactone forms of several statins, including atrovastatin, simvastatin, cerivastatin fluvastatin, pitavastatin and rosuvastatin with respect to metabolic clearance, CYP enzymes involved and drug drug interactions. A remarkable increase in metabolic clearance was noted for all lactones compared with all acids except for pitavastatin lactone. The metabolic clearances of the atrovastatin, simvastatin, cerivastatin, fluvastatin and rosuvastatin lactones were 73-, 70-, 30-, 7- and 64-fold higher, respectively, than those of the corresponding acids. CYP2Cs were critically involved in the metabolism of cerivastatin, fluvastatin and pitavastatin acids. In contrast, CYP2Cs were not involved in the metabolism of the corresponding lactones and CYP3A4 was mainly involved. Moreover, a substantial difference in the metabolic inhibition of statins was found between acids and lactones. Overall, the study demonstrates that CYP-mediated metabolism of lactones is also a common metabolic pathway for statins and that the CYP3A4-mediated metabolism of the lactone forms clearly will need to be taken into account in assessing mechanistic aspects of drug-drug interaction involving statins. PMID- 15801540 TI - Use of a cocktail of probe substrates for drug-metabolizing enzymes for the assessment of the metabolic capacity of hepatocyte preparations. AB - A cocktail of the following probe substrates for human drug-metabolizing enzymes was used to characterize hepatocyte preparations: phenacetin (for CYP1A2), diclofenac (CYP2C9), diazepam (CYP2C19), bufuralol (CYP2D6), midazolam (CYP3A4/5) and 7-hydroxycoumarin (for glucuronidation and sulphation). The cocktail was incubated with cryopreserved human, dog or minipig hepatocytes or with freshly prepared rat hepatocytes. Sample analysis was performed by liquid chromatography tandem mass spectrometry (LC-MS/MS) in an Open Access environment that allowed less experienced MS operators to login, submit and analyse sample sets using predefined settings without the immediate attendance of an experienced analyst. Intrinsic clearances (CLint) were calculated from the disappearance of the compounds from the incubations. Initially, the cocktail used for human, rat and dog hepatocyte incubations contained 7-ethoxycoumarin instead of 7 hydroxycoumarin. However, 7-ethoxycoumarin had an inhibitory effect on the metabolism of phenacetin. The highest CLint estimated with human and dog hepatocytes was observed for 7-hydroxycoumarin. For rat and minipig hepatocytes, the highest CLint was observed for bufuralol. In incubations with dog and minipig hepatocytes, the lowest CLint was seen with diclofenac, whereas for human and rat hepatocytes, the lowest value was observed with diazepam and phenacetin, respectively. When the cocktail was incubated together with human hepatocytes and 1 microM ketoconazole, the CLint of midazolam was decreased to about 7.5% of the control value, whereas the metabolism of the other cocktail compounds was virtually unaffected by this CYP3A inhibitor. It is suggested that a cocktail of specific human probe substrates for drug-metabolizing enzymes can be used routinely for the determination of the metabolic capacity of hepatocyte preparations in order to ensure the quality and reproducibility of experiments. Moreover, a cocktail of specific probe substrates can also be a useful tool for studies on enzyme inhibition. PMID- 15801542 TI - Role of human cytochrome P450 3A4 in the metabolism of DA-8159, a new erectogenic. AB - The purpose of this paper was to characterize cytochrome P450 (CYP) enzymes involved in N-dealkylation of a new oral erectogenic, DA-8159 to DA-8164, a major circulating active metabolite, in human liver microsomes and to investigate the inhibitory potential of DA-8159 on CYP enzymes. CYP3A4 was identified as the major enzyme responsible for DA-8159 N-dealkylation to DA-8164 based on correlation analysis and specific CYP inhibitor and antibody-mediated inhibition study in human liver microsomes, and DA-8159 metabolism in cDNA expressed CYP enzymes. There is the possibility of drug-drug interactions when prescribing DA 8159 concomitantly with known inhibitors or inducers of CYP3A4. DA-8159 was found to be only a very weak inhibitor of eight major CYPs (1A2, 2A6, 2C8, 2C9, 2C19, 2D6, 2E1 and 3A4), the largest inhibition occurring against CYP2D6 (IC5o 67.7 microM) in human liver microsomes. Drug-drug interactions would not be predicted on the basis of DA-8159 inhibiting the metabolism of coadministered drugs. PMID- 15801543 TI - In vitro metabolism of gefitinib in human liver microsomes. AB - The in vitro metabolism of gefitinib was investigated by incubating [14C] gefitinib, as well as M537194, M387783 and M523595 (the main metabolites of gefitinib observed in man), at a concentration of 100 microM with human liver microsomes (4 mg ml(-1)) for 120 min. These relatively high substrate and microsomal protein concentrations were used in an effort to generate sufficient quantities of metabolites for identification. HPLC with ultraviolet light, radiochemical and mass spectral analysis, together with the availability of authentic standards, enabled quantification and structural identification of a large number of metabolites. Although 16 metabolites were identified, metabolism was restricted to three regions of the molecule. The major pathway involved morpholine ring-opening and step-wise removal of the morpholine ring and propoxy side chain. O-demethylation of the quinazoline methoxy group was a quantitatively less important pathway, in contrast to the clinical situation, where O-desmethyl gefitinib (M523595) is the predominant plasma metabolite. The third metabolic route, oxidative defluorination, was only a minor route of metabolism. Some metabolites were formed by a combination of these processes, but no metabolism was observed in other parts of the molecule. Incubation of gefitinib produced ten identified metabolites, but the use of the three main in vivo metabolites as additional substrates enabled a more comprehensive metabolic pathway to be constructed and this has been valuable in supporting the more limited data available from the human in vivo study. PMID- 15801544 TI - In vitro metabolism of quazepam in human liver and intestine and assessment of drug interactions. AB - The study was carried out to identify and characterize kinetically the cytochrome P450 (CYP) enzymes responsible for the major metabolite formation of quazepam. In in vitro studies using human liver and intestinal microsomes and cDNA-expressed human CYP and FMO isoenzymes, quazepam was rapidly metabolized mainly by CYP3A4 and to a minor extent by CYP2C9, CYP2C19 and FMO1 to 2-oxoquazepam (OQ), which was then further biotransformed to N-desalkyl-2-oxoquazepam (DOQ) and to 3 hydroxy-2-oxoquazepam (HOQ) mainly by CYP3A4 and CYP2C9. CYP3A4 is the enzyme predominantly responsible for all the metabolic pathways of quazepam. Itraconazole inhibited the formation of OQ from quazepam, HOQ from OQ and DOQ from OQ in human liver microsomes with Ki values of 8.40, 0.08 and 0.39 microM, respectively. However, the Ki for OQ formation was greater than the peak plasma itraconazole concentration following a clinically relevant 200-mg oral dose to healthy volunteers. In addition, CYP2C9 and CYP2C19 inhibitors failed to inhibit OQ formation from quazepam. In conclusion, clinically relevant drug interaction with CYP inhibitors seem unlikely for the major metabolic pathway of quazepam to OQ. PMID- 15801545 TI - Identification of the cytochrome P450 enzymes involved in the metabolism of domperidone. AB - The objective was to identify the major cytochrome P450 enzyme(s) involved in the metabolism of domperidone. Experiments were performed using human liver microsomes (HLMs), recombinant human cytochrome P450 enzymes, cytochrome P450 chemical inhibitors and monoclonal antibodies directed against cytochrome P450 enzymes. Four metabolites were identified from incubations performed with HLMs and excellent correlations were observed between the formation of domperidone hydroxylated metabolites (M1, M3 and M4), N-desalkylated domperidone metabolite (M2) and enzymatic markers of CYP3A4/5 (r2 = 0.9427, 0.951, 0.9497 and 0.8304, respectively). Ketoconazole (1 microM) decreased the formation rate of M1, M2, M3 and M4 by 83, 78, 75 and 88%, respectively, whereas the effect of other inhibitors (quinidine, furafylline and sulfaphenazole) was minimal. Important decreases in the formation rate of M1 (68%), M2 (64%) and M3 (54%) were observed with anti-CYP3A4 antibodies. Formation of M1, M2 and M3 in HLMs exhibited Michaelis-Menten kinetics (Km: 166, 248 and 36 microM, respectively). Similar Km values were observed for M1, M2 and M3 when incubations were performed with recombinant human CYP3A4 (Km: 107, 273 and 34 microM, respectively). The data suggest that CYP3As are the major enzymes involved in the metabolism of domperidone. PMID- 15801546 TI - Incorporation of xenobiotic carboxylic acids into lipids by cultured 3T3-L1 adipocytes. AB - The study was established to assess the potential for a variety of xenobiotic aromatic carboxylic acids to be incorporated into glycerolipids. The 14C-labelled xenobiotic acids were included in incubations of cultured 3T3-L1 adipocytes under defined conditions. Lipids were extracted and identified by TLC and radioscanning. Ibuprofen, 4-(2,4-dichlorophenoxy)-butanoic acid (2,4-DB), 4-(2 methyl-4-chlorophenoxy)-butanoic acid (MCPB) and 2-(2-methyl-4-chlorophenoxy) propanoic acid (MCPP) (all 0.5 mM) were incorporated into lipid extracts at rates of 220, 227, 199 and 21 pmol microg(-1) phospholipid/h, respectively. 2,4 Dichlorophenoxyacetic acid (2,4-D), indomethacin, naproxen and fluroxypyr were incorporated at rates lower than MCPP or not at all. The incorporation of acids was first order to at least 1 mM acid (except MCPB: 300 microM). Triacylglycerol analogues were the major products with incorporation into diacylglycerol and phosphatidylcholine also observed. After digestion with pancreatic lipase, ibuprofen-containing triacylglycerol was unusual in that the main product was the monoacylglycerol analogue, suggesting that esterification had been at the sn-2 position. Incubation with cultured 3T3-L1 adipocytes is a useful and easy method to assess whether xenobiotic compounds can be incorporated into glycerolipids; of eight acids assessed, four (of which three have not previously been reported) were shown to form xenobiotic triacylglycerols. PMID- 15801547 TI - Triclabendazole biotransformation and comparative diffusion of the parent drug and its oxidized metabolites into Fasciola hepatica. AB - Triclabendazole (TCBZ) is an halogenated trematodicidal benzimidazole compound extensively used in veterinary medicine. It is active against immature and adult stages of the liver fluke Fasciola hepatica. Free and conjugated TCBZ metabolites have been identified in the bile of treated sheep. The experimental aims were to characterize the in vitro patterns of TCBZ biotransformation both in the animal host (sheep liver microsomes) and target parasite (F. hepatica microsomal preparation); and to compare the ex vivo diffusion of TCBZ parent drug and its oxidized metabolites (TCBZ sulphoxide [TCBZSO], TCBZ sulphone [TCBZSO2], and TCBZ hydroxy derivatives) into F. hepatica. Additionally, the octanol-water partition coefficients for TCBZ and all its metabolites were estimated as an indicator of the relationship between drug lipophilicity and diffusion into the target parasite. Drug/metabolites concentrations were quantified by HPLC after sample clean up and a solvent-mediated chemical extraction. Sheep liver microsomes metabolized TCBZ into its sulphoxide and sulphone metabolites after 30 min of incubation. The rate of TCBZ sulphoxidation in the liver was significantly greater (p < 0.01) than that observed for the sulphonation of TCBZSO. The trematode parasite oxidized TCBZ into its sulphoxide metabolite after 60 min of incubation at a metabolic rate of 0.09 nmol min(-1) mg protein(-1). TCBZ and all its oxidized metabolic products were recovered from F. hepatica as early as 15 min after their ex vivo incubation in a Kreb's Ringer Tris buffer. However, the diffusion of the hydroxy-derivatives into the fluke was lower than that observed for TCBZ, TCBZSO and TCBZSO2. There was a high correlation (r=0.82) between drug lipophilicity (expressed as octanol-water partition coefficients) and drug availability measured within the parasite. Unlike the uptake pattern previously observed for albendazole, the parent TCBZ and its sulphoxide and sulphone metabolites showed a similar ability to penetrate into the trematode parasite. Understanding the relationship between TCBZ metabolism, the relative pharmacological potency of its metabolic products and their ability to reach the target parasite may be critical to optimize its flukicidal activity, particularly when TCBZ resistant flukes have been already isolated in the field. PMID- 15801548 TI - Renal elimination of a novel and potent alphavbeta3 integrin antagonist in animals. AB - Compound A (3-{2-oxo-3-[3-(5,6,7,8-tetrahydro-[1,8]napthyridin-2-yl)propyl] imidazolidin-1-yl}-3(S)-(6-methoxy-pyridin-3-yl)propionic acid), a hydrophilic zwitter-ion, is a potent and selective alphavbeta3 integrin antagonist currently under clinical development for the treatment of osteoporosis. The mechanism of renal excretion of compound A was investigated using a combination of in vivo and in vitro approaches. In rats, renal excretion of compound A involved tubular secretion; ratios between renal clearance, corrected for unbound fraction in plasma (CLr,u) and glomerular filtration rate (GFR) were greater than unity (2 5). The tubular secretion of compound A was saturable at high plasma levels (> 26 microM), and was inhibited significantly, although modestly (about twofold) by relatively high plasma concentrations of the organic anion PAH (160 microM) and the cation cimetidine (about 400 microM), but not by the P-gp inhibitor quinidine (about 50 microM). However, compound A (about 100 microM) had a minimal effect on CLr/GFRs for cimetidine and PAH. In rhesus monkeys, renal elimination of compound A also involved tubular secretion, with a CLr,u/GFR ratio of about 30. The renal secretion of compound A was not affected by either cimetidine (about 120 microM) or PAH (about 80 microM). Similarly, compound A (about 40 microM) had a minimal effect on the renal tubular secretion of both cimetidine and PAH. At the doses studied, neither rat nor monkey plasma protein binding of compound A, cimetidine or PAH was affected in the presence of each other. In vitro transport studies showed that compound A was not a substrate for P-gp in the Caco-2, human MDR1 and mouse mdr1a transfected LLC-PK1 cell lines. In an uptake study using rOAT1 and rOAT3 transfected HEK cell lines, compound A was shown to be a substrate for rat OAT3 (Km= 15 microM), but not rat OAT1. The results suggest that the tubular secretion of compound A is not mediated by P-gp, but rather is mediated, at least in part, via the organic anion transporter OAT3, the renal transporter shown to be capable of transporting both the organic anion PAH and the organic cation cimetidine. Although there is a possibility for pharmacokinetic interactions between compound A and substrates or inhibitors of OAT3, at the renal excretion level, the magnitude of interaction would likely be modest in humans at clinically relevant doses. PMID- 15801549 TI - Characterization and quantification of metabolites of racemic ketoprofen excreted in urine following oral administration to man by 1H-NMR spectroscopy, directly coupled HPLC-MS and HPLC-NMR, and circular dichroism. AB - The identity of the human metabolites of ketoprofen (2-(3-benzoylphenyl) propanoic acid) excreted via urine was investigated after a single oral dose of the racemic drug. Drug metabolites were concentrated and partially purified from urine using solid-phase extraction chromatography before separation and identification by directly coupled HPLC-MS and HPLC-NMR. The metabolites identified were the ester glucuronides of the parent drug and its phase I metabolites, 2-[3-(3-hydroxybenzoyl)phenyl]-propanoic acid, 2-[3-(4 hydroxybenzoyl)phenyl]-propanoic acid and 2-[3-(hydroxy(phenyl)methyl)phenyl] propanoic acid, the latter formed by reduction of the ketone group of ketoprofen. In addition, two novel minor metabolites were identified as the ether glucuronides of 2-[3-(3-hydroxybenzoyl)phenyl]-propanoic acid and 2-[3-(4 hydroxybenzoyl)phenyl]-propanoic acid. These conjugates were all observed as diastereoisomeric pairs of unequal proportions. Purification of these metabolites by preparative chromatography allowed stereochemistry assignments. Metabolites were quantified by 1H-NMR spectroscopy after spectral simplification achieved by hydrolysis of the conjugates. PMID- 15801550 TI - Community-based participatory research to prevent substance abuse and HIV/AIDS in African-American adolescents. AB - Adolescence is a time for exploration and risk-taking; in today's urban environment, with the twin threats of substance abuse and HIV/AIDS, the stakes are particularly high. This paper describes a community-based participatory research project to design, implement, and evaluate a faith-based substance abuse and HIV/AIDS prevention program for African-American adolescents. A coalition of university-based investigators and African-American church member stakeholders collaborated on all aspects of Project BRIDGE, the 3-year intervention to reduce substance abuse and HIV/AIDS in African-American adolescents. Our results support the use of community-based participatory research to create desirable change in this setting. Adolescents who participated in Project BRIDGE reported significantly less marijuana and other drug use and more fear of AIDS than a comparison group. Project BRIDGE has been designated an official ministry of the church and the program has been extended to others in the larger metropolitan community. The church now has a well-trained volunteer staff University faculty developed skills in negotiating with community-based settings. The coalition remains strong with plans for continued collaborative activities. PMID- 15801551 TI - Addressing the realities [correction of realties] of health care in northern aboriginal communities through participatory action research. AB - To address concerns about disruptions in the continuity of health care delivered to residents in three remote aboriginal communities in northern Ontario, Canada, the local health authority initiated a study in collaboration with the department of Health Canada responsible for ensuring that aboriginal reserves receive mandatory health services, and an inter-disciplinary team of researchers from two universities. The study focussed on the delivery of oncology, diabetes and mental health care, specifically, as well as systems issues such as recruitment and retention of health human resources and financial costs. The paper discusses the procedures involved, the benefits derived and the challenges encountered in doing this as a community driven participatory action research project. It also summarizes the findings that led to community formulated policy and program recommendations. PMID- 15801552 TI - Use of community-based participatory research in preparing low income and homeless minority populations for future HIV vaccines. AB - We conducted Community-Based Participatory Research (CBPR), using a qualitative focus group design, to assess factors that might impact participation of high risk impoverished adults in future HIV Vaccine Trials (HIVVTs). The participants were 40 homeless and low-income adults recruited from subsidized apartments and homeless shelters in Los Angeles. Findings revealed that the participants expressed both concerns and interest in future HIVVTs. Concerns centered on the impact of the vaccine on their physical health, the possibility of seroconverting and its associated stigma. While distrust of the government was pervasive, the participants were interested in receiving more information about the vaccine from the researchers. They also wished to have their voices heard by the researchers early in the design of the vaccines. Motivating factors were also discovered, and included altruism, compensation and access to care. Perception that risk behaviors might increase among some as a result of participation in a future HIVVT was likewise revealed. Implications of the study reveal that while impoverished populations are interested in participating in future HIVVTs, the researchers must address concerns early on. Moreover, the importance of ongoing education and counseling to warn about hazards of engaging in risky behavior while participating in a future HIVVT was critical. PMID- 15801553 TI - Social audit of provincial health services: building the community voice into planning in South Africa. AB - Social audits of health services in three South African provinces (Limpopo 2001, Gauteng 2003 and Eastern Cape 2001) demonstrated a fusion of participatory research, qualitative data collection, epidemiological surveys and analysis, and socialising of evidence for action. The social audit in each province involved the community in covering eight principles of service delivery. In the particular case of Gauteng province, householders answered a questionnaire on public perceptions of government health services. In each sentinel community, the results were returned for discussion in focus groups, made up of a sample that had answered the household questionnaire. Institutional reviews of health facilities and interviews with health workers in those facilities gave the government side of the story. Five actions came from the community-based focus groups, all accepted by the provincial government. Firstly, redesigned communication strategies will aim to reach those with lower levels of education. The second set of actions involves rebuilding the culture of care. Thirdly, the public knowledge of ways to complain was closely related to levels of public satisfaction. Leadership in the health services turned out to be a fourth area for development. And finally, related to this, is the longer-term challenge of establishing and reinforcing community consultation mechanisms. PMID- 15801554 TI - Cardiovascular health among Asian Indians (CHAI): a community research project. AB - The object of this research was to assess cardiovascular (CV) risks in Asian Indians in California. We conducted eight focus groups and a pilot survey using community-based participatory research (CBPR) methods. Focus groups were held in six communities. Surveys were conducted by telephone or in person in areas selected for high population densities of Asian Indians. We selected focus group subjects by snowball sampling (n = 57). We held six English and two Punjabi groups. We used a surname-based phone list from three area codes for telephone interviews (n = 254). We added 50 in-person interviews for comparison (total n = 304) and did 50 interviews in Punjabi. We held community meetings for dissemination. Focus groups discussed CV risks; themes developed aided survey development. In-person and telephone surveys were feasible. Telephone surveys were more gender-balanced and people more often answered alcohol, tobacco, and income questions. Self-reported prevalences for hypertension, hypercholesterolemia, and diabetes were 20.4, 35.3 and 10.6%, respectively. Only 11.9% of persons reported ever smoking cigarettes. It was concluded that CBPR methods were effective in this exploratory study assessing CV risks in Asian Indians. Hypertension, high cholesterol, and diabetes were more prevalent in participants than the population average; other risk factors were less common (tobacco). PMID- 15801555 TI - Realidad Latina: Latino adolescents, their school, and a university use photovoice to examine and address the influence of immigration. AB - Over the past 10 years, growth of the Latino population in the United States has been most rapid in North Carolina. Project Realidad Latina (Latino Reality) was a qualitative exploratory study conducted to gain insight into the immigration experiences of 10 newly-arrived Latino adolescents living in rural North Carolina (NC). The study followed a community-based participatory research (CBPR) approach and used the photovoice method. Over a one-year period, adolescents partnered with public health practitioners and researchers in: generating photo assignments, taking photographs based on these assignments, using the photographs for photo-discussions, and defining themes based on these photo-discussions. A photograph exhibition and community forum raised awareness among local decision makers and community members of the issues and assets of Latino adolescents and initiated a process toward change. From the participants' words and photographs emerged contextual descriptions of issues that both challenged and facilitated their adaptation and quality of life in their school and community. Likewise, implications from the findings and the nature of the CBPR approach for future Latino adolescent health intervention research are presented. PMID- 15801556 TI - Health professional faculty perspectives on community-based research: implications for policy and practice. AB - Community-based research (CBR) has become central to the understanding and elimination of health disparities within the USA and across the globe. The authors sought to determine the perspectives of health professional faculty on the factors affecting their involvement in CBR and the extent of community participation in that research. Faculty from 18 health professional schools in the USA identified by their deans as being leaders in CBR completed a written survey. Respondents reported that between 5-10% of faculty in their schools were involved in CBR. Public perception of the university, familiarity with community based organization leaders and institutional leadership were cited as the most significant factors contributing to a school's involvement in CBR. Long-term community relationships, recognition in tenure and promotion policies and access to funding were cited as factors that support faculty in conducting CBR. The authors conclude that a more significant investment of public and private funds, the development of interdisciplinary institutional structures for community partnerships and a more inclusive definition of scholarship are needed to achieve a central role for CBR in efforts to understand and eliminate health disparities. PMID- 15801557 TI - Funding community-based participatory research: lessons learned. AB - The California Breast Cancer Research Program (CBCRP) was created to fund innovative breast cancer research specifically addressing the needs of women in California. Beginning in 1997, the Program launched the Community Research Collaboration (CRC) Program, a Community-Based Participatory Research (CBPR) program intended to foster community-researcher collaboration on all aspects of the research process, essentially placing the community in the center of the research paradigm. The CBCRP conducted a process evaluation of the CRC Program to assess success and identify areas for improvement. The evaluation included community-researcher collaborations and the award process. The evaluation identified successes that speak to the effectiveness of the collaboration concept: empowering women to formulate and initiate research; involving underserved and hard-to-reach populations; addressing important and useful research questions; increasing communities' skills and expertise, and enabling lasting collaborations. The greatest weakness identified was the involvement of the broader community (beyond the group/organization involved) in these projects. The evaluation identified strengths in CBCRP's award process, including technical assistance and feedback, the emphasis on collaboration in the review process, and awarding funds directly to community groups. Barriers included: power imbalances due to the community's lack of experience in the dominant research funding culture; funding limits and award delays; and the increased service demand beyond the funding limits of the award that is created. The CBCRP has been able to incorporate many of the evaluation findings to improve the CRC Program. PMID- 15801558 TI - Partnership for evaluation of the Bilingual Bicultural Service Demonstration Program: merging public health research and practice. PMID- 15801559 TI - Getting To Outcomes: a community-based participatory approach to preventive interventions. PMID- 15801560 TI - Community-based participatory research on smoking cessation among Chinese Americans in Flushing, Queens, New York City. PMID- 15801561 TI - Poder es Salud/Power for Health: an application of the community health worker model in Portland, Oregon. PMID- 15801562 TI - The Healthy Weigh/El Camino Saludable: a community campus partnership to prevent obesity. PMID- 15801563 TI - Health literacy: expanding practitioners' horizons through collaborative research. PMID- 15801564 TI - Participatory research in primary care. PMID- 15801565 TI - Project 'M'--women living with cancer: a participatory-action evaluation of a demonstration project. PMID- 15801566 TI - The neurobiology of tobacco dependence: a preclinical perspective on the role of the dopamine projections to the nucleus accumbens [corrected]. AB - It is now widely accepted that nicotine is the primary addictive component of tobacco smoke and that a majority of habitual smokers find it difficult to quit smoking because of their dependence upon this component of the smoke. However, although nicotine replacement therapy elicits a clinically valuable and significant improvement in the number of quit attempts that are ultimately successful, its efficacy remains disappointingly low. This review considers some of the reasons for this problem. It focuses on the hypothesis that stimulation of the dopamine (DA) projections to the medial shell and the core of the nucleus accumbens play complementary roles in the development of nicotine dependence. The hypothesis proposes that increased extra-synaptic DA in the medial shell of the accumbens confers hedonic properties on behaviors, such as smoking, which deliver nicotine, and thereby increase the probability that the response is learned. It also summarizes the evidence that the primary role of the increased DA overflow, observed in the accumbal core of nicotine-pretreated individuals, challenged with nicotine, is the attribution of incentive salience to cues associated with delivery of the drug and the transition to Pavlovian responding to these conditioned stimuli. The review argues that sensitization of the DA projections to the accumbal core, and the behaviors that depend upon this process, play a pivotal role in the maintenance of the tobacco smoking habit and that it is this component of the dependence that is inadequately addressed by nicotine replacement therapy. PMID- 15801567 TI - The impact of smoking cessation on objective and subjective markers of sleep: review, synthesis, and recommendations. AB - Sleep disturbance is commonly reported as a prominent subjective symptom by quitting smokers. However, little research on this issue has used objective measures of sleep quality. Previous research has relied mainly on retrospective report of sleep disturbance, with few studies investigating sleep during the initial period after quitting tobacco use. Studies that have used objective measurements suggest that sleep fragmentation is a common occurrence during the withdrawal period. In sleep medicine, sleep disturbance is viewed as a consequence of frequent arousals and is now considered to have particularly deleterious daytime consequences, including sleepiness and dysphoric mood. Recent work also indicates that such awakenings affect the cardiovascular system by providing repetitive bursts of sympathetic nervous system activation, possibly contributing to elevated levels of cardiovascular and cerebrovascular morbidity. Pharmacological treatments designed to facilitate smoking cessation are ineffective for sustained abstinence in many smokers, which may be related to sleep disturbance. Indeed, preliminary evidence suggests that the administration of nicotine replacement therapy (NRT) or bupropion can result in disrupted sleep, particularly in women. However, to better understand the role that nicotine withdrawal and bupropion or NRT treatment, independently and in combination, might play in sleep disturbance, it is necessary to develop a better understanding of the nature of the sleep disturbance than can be provided by self report. This is particularly important for the development of treatment approaches targeted to ameliorate sleep disruption as part of an overall smoking cessation strategy. The present review seeks to report the current state of knowledge based on extant findings and argues for the need to conduct more detailed polysomnographic investigations of the potentially vicious cycle of smoking cessation leading to sleep disturbances that may prove iatrogenic to sustained cessation. PMID- 15801568 TI - Assessing internal tobacco industry knowledge of the neurobiology of tobacco dependence. AB - The recent availability of internal tobacco industry documents provides a significant resource for evaluating industry understanding of the pharmacological, psychosocial, and behavioral mechanisms underlying tobacco dependence. In this study, we catalog the range of efforts undertaken by tobacco manufacturers seeking knowledge of these mechanisms. Some areas of industry research, such as cellular and molecular studies of nicotine and its effects, are widely available in the open literature. Of greater interest are internal research projects that have demonstrated direct influence on product development. These include studies of smoker psychology and behavior, evoked-response studies of tobacco-delivered nicotine, the effects of sensory perception, dose-related effects, and the development of nicotine analogs and synergists. Our findings suggest extensive industry knowledge of mechanisms that determine smoker perception and behavior, and application of this knowledge in product development, including control of sensory response, uptake of nicotine, and product effects. Independent research recently has begun to consider the contributions of tobacco product ingredients and design factors to the determination of risk, severity, and prevalence of addiction. However, the application of these findings to cessation and treatment efforts is still quite limited. We conclude that clinical research would greatly benefit from further examination of the decades of knowledge accumulated by tobacco manufacturers. PMID- 15801569 TI - Human functional neuroimaging in nicotine and tobacco research: basics, background, and beyond. AB - Modern functional neuroimaging techniques allow nicotine and tobacco researchers to investigate the neurobiological basis of addiction in humans. We introduce the methods and measures of the following neuroimaging techniques: Electroencephalography and event-related cortical potentials, positron emission tomography, and functional magnetic resonance imaging. We outline strengths and limitations across modalities and describe new and emerging technologies. We provide summaries of recent neuroimaging findings in the field of nicotine and tobacco research for neurochemistry, smoking and nicotine administration, craving and cue-reactivity, cognitive and affective information processing, and tobacco withdrawal. We address limitations of studies to date and identify opportunities for future research. PMID- 15801570 TI - Effects of nicotine and smoking on event-related potentials: a review. AB - Event-related potentials can serve as an adjunct to reaction time in elucidating the effects of nicotine on rapid human information processing. We review the literature related to nicotine and event-related potentials. Although evidence indicates that, in the visual modality, nicotine enhances early perceptual processing and in certain instances speeding stimulus evaluation, the bulk of nicotine's effects seem to be on enhancing response preparation and response execution. PMID- 15801571 TI - Effects of nicotine on brain responses to emotional pictures. AB - Given that nicotine reduces negative affect, one would expect nicotine to have different effects on brain responses to emotionally negative stimuli than it does on responses to emotionally neutral or positive stimuli. However, no studies have assessed this possibility. The present study assessed the effects of nicotine patch versus placebo patch on brain event-related potential (ERP) responses to emotion-inducing negative, positive, and neutral color pictures in 16 smokers in a double-blind, counterbalanced, within-subjects design. The study included four experimental sessions. After overnight smoking deprivation (12 hr or more), active nicotine patches were applied to participants during one of the first two sessions and during one of the last two sessions. Placebo patches were applied during the other two sessions. Nicotine reduced frontal ERP processing voltage negativity (from 144-488 ms poststimulus onset) evoked by viewing emotionally negative pictures to a greater extent than it did when emotionally neutral pictures were viewed, whereas it had no effect on processing negativity evoked by positive pictures. Nicotine also enhanced P390 amplitudes evoked by emotionally negative pictures more than it did when emotionally neutral and positive pictures were viewed. Across picture types, nicotine (relative to placebo) reduced N300 amplitude (more at anterior and dorsal sites) and increased P390 amplitude. Overall, nicotine influenced ERPs to emotionally neutral and positive pictures less than it did to negative pictures. PMID- 15801572 TI - Determinants of salivary cotinine levels among current smokers in Mexico. AB - The present study describes salivary cotinine levels and their relationship to cigarettes smoked per day in Mexican smokers. Using a sampling strategy based on the number of cigarettes per day, we recruited 1,222 smokers from Mexico City and the state of Morelos in Mexico during 1999. Smoking behaviors and other factors known to affect nicotine intake and cotinine level were identified in an interview using a standardized questionnaire. Salivary cotinine was measured by capillary gas chromatography with nitrogen-phosphorus detection. We used generalized additive models to describe the relationship between salivary cotinine levels and variables of interest. The mean age of the population was 39.7 years (SD=15.6 years), with a mean cotinine level of 194.7 ng/ml (SD=134.8; range=10.1-767). Participants smoked a mean of 15.5 cigarettes per day (SD=11.3). Salivary cotinine and cigarettes smoked per day were positively related, although the association was not linear, flattening above 20 cigarettes per day. After adjusting for cigarettes per day, we found that significant predictors of cotinine levels included age, body mass index, cigarette producer, and smoking behavior variables. These results may have implications for dosing with nicotine medications to aid smoking cessation in Mexican smokers and suggest that whether the cigarette is labeled light or regular has no relationship to nicotine dose from smoking cigarettes. PMID- 15801573 TI - L-arginine prevents impaired endothelium-dependent cerebral arteriolar dilatation during acute infusion of nicotine. AB - Exogenous treatment with L-arginine has been shown to restore impaired nitric oxide synthase (NOS)-dependent dilatation of peripheral blood vessels during disease states. We have shown that nicotine impairs NOS-dependent arteriolar dilatation in the cerebral circulation. However, the role of L-arginine in impaired responses of cerebral arterioles during infusion of nicotine has not been examined. Thus the goal of the present study was to examine the role of L arginine in nicotine-induced impairment of cerebral arteriolar reactivity. We measured the diameter of pial arterioles in response to NOS-dependent (5' adenosine diphosphate [ADP] and acetylcholine) and NOS-independent (nitroglycerin) agonists before and after infusion of vehicle or nicotine (2 microg/kg/min intravenously for 30 min followed by a maintenance dose of 0.35 microg/kg/min) in the absence or presence of L-arginine (10(-3) M). We found that topical application of L-arginine to cerebral microvessels during infusion of nicotine could prevent impaired NOS-dependent vasodilatation. We suggest that exogenous L-arginine may have a beneficial role in preventing cerebral microvascular dysfunction during exposure to nicotine. PMID- 15801574 TI - A pilot study on voucher-based incentives to promote abstinence from cigarette smoking during pregnancy and postpartum. AB - We report results from a pilot study examining the use of vouchers redeemable for retail items as incentives for smoking cessation during pregnancy and postpartum. Of 100 study-eligible women who were still smoking upon entering prenatal care, 58 were recruited from university-based and community obstetric practices to participate in a smoking cessation study. Participants were assigned to either contingent or noncontingent voucher conditions. Vouchers were available during pregnancy and for 12 weeks postpartum. In the contingent condition, vouchers were earned for biochemically verified smoking abstinence. In the noncontingent condition, vouchers were earned independent of smoking status. Abstinence monitoring and associated voucher delivery was conducted daily during the initial 5 days of the cessation effort, gradually decreased to every other week antepartum, increased to once weekly during the initial 4 weeks postpartum, and then decreased again to every other week for the remaining 8 weeks of the postpartum intervention period. Contingent vouchers increased 7-day point prevalence abstinence at the end-of-pregnancy (37% vs. 9%) and 12-week postpartum (33% vs. 0%) assessments. That effect was sustained through the 24-week postpartum assessment (27% vs. 0%), which was 12 weeks after discontinuation of the voucher program. Total mean voucher earnings across antepartum and postpartum were 397 US dollars (SD=414 US dollars) and 313 US dollars (SD=142 dollars) in the contingent and noncontingent conditions, respectively. The magnitude of these treatment effects exceed levels typically observed with pregnant and recently postpartum smokers, and the maintenance of effects through 24 weeks postpartum extends the duration beyond those reported previously. PMID- 15801575 TI - The effect of a novel smoking system--Accord--on ongoing smoking and toxin exposure. AB - Multiple potentially reduced exposure products (PREPs) are being introduced to the market, yet little is known about how they will be used and what their public health impact might be. To determine the impact of one such PREP--Accord--on ongoing smoking and toxin exposure, 11 smokers of light cigarettes were required to use increasing amounts of Accord (5, 10, and 15 per day) with the option of using their traditional cigarettes. Accord suppressed ongoing cigarettes per day and carbon monoxide (CO), but not cotinine, in a dose-dependent manner. Smoking 15 Accord per day decreased the number of traditional cigarettes smoked by 32% ( 8.6 cigarettes per day) and CO levels by 27% (-5.9 ppm). However, Accord did not function as a perfect (i.e., one to one) substitute for cigarettes because the total number of nicotine products (Accord plus usual brand) per day increased by 24%. Participants believed that Accord was safer than traditional cigarettes but rated Accord as ineffective at suppressing cravings for cigarettes. These findings suggest that use of Accord results in significant decreases in cigarettes smoked and CO exposure. Whether these reductions will translate into health benefits or endure beyond 2 weeks is unknown. Because most PREPs will probably be used along with traditional cigarettes, their net health impact is a function of not only their toxicological profile but also their effect on ongoing smoking. PMID- 15801576 TI - Comorbidity of depression with levels of smoking: an exploration of the shared familial risk hypothesis. AB - Comorbidity of depression and smoking is well recognized, but results from studies that have assessed alternative explanations have varied by the level of smoking and the study method. We examined all 13 etiology models of comorbidity described by Neale and Kendler (American Journal of Genetics, 57, 935-953, 1995) for depression and each of four levels of smoking to shed light on the role that differing definitions might have played in generating the conflicting findings. Data came from 979 young adults aged 26-35 years who participated in an epidemiological cohort study in southeastern Michigan. Respondent and family history data on parental smoking and depression were analyzed using the biometric modeling method for family data, which Rhee and colleagues (Journal of Child Psychology and Psychiatry and Allied Disciplines, 44, 612-636, 2003; Behavior Genetics, 34, 251-265, 2004) have shown to be valid more frequently than traditional prevalence analyses. Results of the biometric model fitting suggested that for ever smoking, the comorbidity with depression may be related to chance or a high liability threshold for smoking only. In contrast, a correlated liabilities model fit the data best for the comorbidity of depression with daily, heavy, and nicotine-dependent smoking. The familial correlations accounted for 73%-95% of the total variance shared between depression and these levels of smoking. These results differ from analyses of these data using a traditional prevalence approach, which found no evidence of shared familial liability. The conflicting findings of the studies that have examined the relationship between smoking and depression may be attributable to differences in definition of the disorders and the methods used to analyze them. PMID- 15801577 TI - Tissue levels of malondialdehyde after passive smoke exposure of rats for a 24 week period. AB - The present study was to evaluate the effects of chronic cigarette smoke exposure on lipid peroxidation in various organ tissues. Sprague-Dawley rats were exposed to passive smoking 2 hr per day, 6 days per week (Monday-Saturday), for 24 weeks. Malondialdehyde levels, as an index of lipid peroxidation, were measured by the thiobarbituric acid assay. Levels were significantly higher in tissues of passive smoke-exposed groups (n=10) compared with normal-bred control groups (n=6), for red blood cells (2.17+/-0.22 vs. 1.80+/-0.39 nmol/mg), lung (1.39+/-0.32 vs. 1.03+/-0.35 nmol/mg), and spleen (1.75+/-0.33 vs. 1.42+/-0.15 nmol/mg); p<.05. No differences in malondialdehyde levels were found in plasma, heart, liver, stomach, and renal tissues. The results suggest that chronic environmental tobacco smoke exposure can increase lipid peroxidation in red blood cells and in lung and spleen tissue. This finding brings further investigative attention to the public health issue of the injurious effects of chronic passive smoke exposure. PMID- 15801578 TI - Subtyping general population smokers not intending to quit by stages to reduce smoking. AB - The present study proposes target groups for preventive measures in smokers not intending to quit based on the intention to reduce smoking using the stages of change concept. Smokers were identified within a representative general population sample (T1; N=4,075) and assessed after 30 months (T2; N=913) and 36 months (T3; N=786). The cross-sectional analyses of the present study included at T2 677 smokers not intending to quit within the next 6 months (cessation precontemplation stage). The prospective analyses were based on 584 of these smokers who were followed up at T3. At T2, 9.1% intended to reduce substantially within the next 6 months (reduction-contemplation stage), 1.2% intended to reduce substantially within the next 4 weeks (reduction-preparation stage), 3.7% had already reduced for up to 6 months (reduction-action stage), and 8.2% had maintained reduction for more than 6 months (reduction-maintenance stage). Subjects in the different reduction stages differed with respect to previous quit or reduction attempts, degree of nicotine dependence, and amount of cigarette consumption. Advanced reduction stages were significantly associated with increased chance of future reduction attempts. Future progress in the cessation stages (OR=4.7, 95% CI=1.5-13.1) and future quit attempts (OR=4.8, 95% CI=1.3 15.2) were significantly more likely for those in the reduction-action stage compared with the reduction-precontemplation stage. Analyses separating the components of the stage measure revealed further need for the validation of the common operational definition. In conclusion, a substantial subgroup of smokers not intending to quit could be subtyped by the reduction stages and might be accessible by smoking-reduction interventions. Our data did not support an undermining effect associated with later reduction stages with respect to future quit attempts and progress within the cessation stages. PMID- 15801579 TI - Instructions about nicotine dose influence acute responses to nasal spray. AB - Effects of substance use are typically assumed to be related to pharmacological actions. However, beliefs about the drug content of a substance may strongly influence subjective and reinforcing responses to that substance (i.e., "placebo" effects). We examined the subjective and reinforcing effects of a nasal spray containing no nicotine as a function of instructions about the nicotine content of the spray (Told Nicotine vs. Told No Nicotine). Smokers (n=49) not interested in quitting smoking abstained overnight prior to a single session in which they were randomly assigned to one of three groups, involving one of the two instructional sets or a group that got no spray. Following dose instructions, subjects in the two spray groups were administered one set of four sprays from the spray bottle and then rated them intermittently on items related to "reward" (e.g., "liking", amount they would pay for more) and other effects. At the same time points, they also rated mood, craving, and withdrawal, and had heart rate and blood pressure measured. Reinforcement was then determined by the number of ad libitum sprays they self-administered during a 20-min period. The no-spray group simply rested quietly during the session, while measures were assessed at the same time points as subjects in the other two groups. Those in the Told Nicotine group reported greater spray ratings of "how much nicotine," "liking," "satisfying," "buzz/head rush," and "similar to smoking" compared with the Told No Nicotine group. Craving decreased more for those Told Nicotine versus those Told No Nicotine, but also decreased more for those Told No Nicotine compared with the no spray group. There were no significant differences in amount they would pay for more sprays, withdrawal, mood, cardiovascular responses, or in spray self-administration. These results show that instructions about the nicotine content of a novel delivery device (nasal spray) can influence self reported spray ratings and reduce craving but are limited with respect to effects on other measures of drug response. PMID- 15801580 TI - Behavioral counseling for reducing children's ETS exposure: implementation in community clinics. AB - The present randomized controlled trial tested the effectiveness of a behavioral counseling program for reducing children's exposure to environmental tobacco smoke (ETS). Counseling was delivered by clinic staff as part of well-child health care services in a community clinic setting. A total of 150 mothers with children aged 4 years or younger were recruited. Parent-reported and children's urinary cotinine measures of ETS exposure were obtained at baseline, 3 months, 6 months (post-test), and 12 months (follow-up). Saliva samples were obtained from mothers who reported quitting smoking, for objective verification by thiocyanate analysis. After baseline, mothers were randomly assigned to a measures-only control condition or an intervention consisting of seven behavioral counseling sessions over 6 months. Counseling included behavioral contracting, self monitoring, problem solving, and positive reinforcement. Results indicated acceptable test-retest reliability and validity of measures. Parent-reported measures indicated that, in both groups, children's exposure to their mothers' tobacco smoke in the home and to all tobacco smoke declined steeply from baseline to 6 months post-test, and remained essentially level during follow-up. Mothers' smoking rates followed the same pattern. Children's urinary cotinine concentrations did not show significant change over time in either group. Findings on the fidelity of treatment implementation suggest that the structure and funding of the community clinic health care system and associated staff turnover and training issues resulted in participants receiving a less efficacious intervention than in our past efficacy trials. Implications for future effectiveness trials are discussed. PMID- 15801581 TI - Recent quitters' interest in recycling and harm reduction. AB - Many smoking cessation attempts are followed by failure or relapse. The optimal time to initiate a new quit attempt is not known. Administrative databases documenting recent use of a pharmacological aid for smoking cessation provide access to a population of smokers recently in the action phase of quitting. This study describes interest in further treatment among this population. A total of 2,340 smokers from the Minneapolis Veterans Administration Medical Center received prescriptions for a smoking cessation aid during an 18-month period. We conducted a cross-sectional survey of a random sample of this population a minimum of 3 months following the prescription, using a structured telephone interview (N=391). The response rate was 75.8%. The 1-month point-prevalent abstinence rate was 19.7%. Of continuing smokers, 98% were willing to make another quit attempt--50% immediately, and 28% within 1 month. There was no relationship between time since the last quit attempt and interest in trying again immediately. Some 95% of continuing smokers were willing to try a reduction strategy. Of these, 82.7% were interested in using nicotine replacement therapy (NRT) to accomplish this goal. Most cessation programs do not systematically approach participants who relapse. These data suggest that this population would welcome further help in quitting or reducing smoking shortly following failure, and that smokers do not commonly relapse to a precontemplation stage of change. PMID- 15801582 TI - Beaked whales echolocate on prey. AB - Beaked whales (Cetacea: Ziphiidea) of the genera Ziphius and Mesoplodon are so difficult to study that they are mostly known from strandings. How these elusive toothed whales use and react to sound is of concern because they mass strand during naval sonar exercises. A new non-invasive acoustic ording tag was attached to four beaked whales(two Mesoplodon densirostris and two Ziphius cavirostris) and recorded high-frequency clicks during deep dives. The tagged whales only clicked at depths below 200 m, down to a maximum depth of 1267 m. Both species produced a large number of short, directional, ultrasonic clicks with significant energy below 20 kHz. The tags recorded echoes from prey items; to our knowledge, a first for any animal echolocating in the wild. As far as we are aware, these echoes provide the first direct evidence on how free-ranging toothed whales use echolocation in foraging. The strength of these echoes suggests that the source level of Mesoplodon clicks is in the range of 200-220 dB re 1 microPa at 1 m. This paper presents conclusive data on the normal vocalizations of these beaked whale species, which may enable acoustic monitoring to mitigate exposure to sounds intense enough to harm them. PMID- 15801583 TI - Cache protection strategies by western scrub-jays (Aphelocoma californica): hiding food in the shade. AB - In the presence of conspecifics, food-caching western scrub-jays (Aphelocoma californica) implement a variety of strategies to reduce the chances of cache theft. This experiment aimed to determine whether the jays could exploit an environmental variable, the level of ambient light, to reduce the transfer of visual information to potential pilferers. Each jay was allowed to cache non degradable food in two trays, one of which was well lit, whereas the other was in shadow. In some trials the birds cached in private and in others they were observed; however, they always recovered their caches in private. When observed the jays preferentially cached in the shaded tray, whereas both trays were used equally when caching in private. By caching in shaded sites, the quality and transfer of visual information available to the observer may be reduced, thereby making the location of cache sites less certain. These results suggest that western scrub-jays may selectively cache in the shade as a strategy to reduce the chance of cache theft by observing conspecifics. PMID- 15801584 TI - Evolutionary distinctiveness of the extinct Yunnan box turtle (Cuora yunnanensis) revealed by DNA from an old museum specimen. AB - Cuora yunnanensis is an extinct turtle known from 12 specimens collected from Yunnan, China, before 1908. We used ancient DNA methods to sequence 1723 base pairs of mitochondrial DNA from a museum specimen of C. yunnanensis. Unlike some rare 'species' recently described from the pet trade, C. yunnanensis represents a lineage that is distinct from other known turtles. Besides C. yunnanensis, two other valid species (C. mccordi, C. zhoui) are unknown in the wild but are supposedly from Yunnan. Intensive field surveys for surviving wild populations of these critically endangered species are urgently needed. PMID- 15801585 TI - Host resistance and the evolution of kin recognition in polyembryonic wasps. AB - Recognition of relatives is considered a key factor in the evolution of sociality as it ensures that the benefits of altruism flow to those who share the altruist's genes. However, theory predicts that genetically based recognition systems will not persist if the only selection maintaining them derives from the recognition system itself. Kin-recognition systems, therefore, are hypothesized to involve genetic variation maintained by other functions. Polyembryonic wasps are parasites of moth larvae that clonally produce large numbers of offspring and two morphologically distinct castes. Some embryos develop into reproductive larvae that mature into adult wasps, whereas others develop into soldier larvae whose function is defence. Soldiers from Copidosoma floridanum distinguish relatives from non-relatives on the basis of relatedness. Here, we report that the recognition cues used by soldiers derive from the extraembryonic membrane, which also protects these parasites from the host's immune response. This suggests that the kin-recognition system used by C. floridanum may be maintained in part by selection for resistance against the host. PMID- 15801586 TI - Sex-biased predation by polecats influences the mating system of frogs. AB - In agile frogs, Rana dalmatina, an increase in male-biased operational sex ratio and in male abundance results in the emergence of alternative male mating behaviour in the form of searching. As a consequence, females are coerced into mating with multiple males, which in turn increases the level of conflict between the sexes. Selective predation on males by the European polecat, Mustela putorius, decreases the occurrence of polyandry. In ponds visited by polecats, the sex ratio is less male biased than in ponds where polecats are absent. As a result most males call to attract females and fewer males actively search for females. Females are able to choose between calling males and mate with a single male. Thus, predation by polecats is found to influence sex ratio, male abundance and sexual conflict in a frog mating system, restricting the opportunity for multiple mating. PMID- 15801587 TI - Extreme repeated mating as a counter-adaptation to sexual conflict? AB - The Australian scaly cricket, Ornebius aperta, can copulate over 50 times with the same partner; the benefits of such extreme repeated copulation are unclear. We support the hypothesis that repeated copulation increases insemination success, as the number of sperm transferred increases with each spermatophore. This probably increases paternity for males, as on average a female mates with over 40 males. Despite intense sperm competition each ejaculate has only a few hundred sperm, orders of magnitude less than in related crickets. We show that all sperm are transferred from each spermatophore in the few seconds before a female removes and eats it. Repeated copulation increases effective copulation duration while a small ejaculate ensures that this strategy is not excessively costly. Thus repeated copulation in these crickets may have arisen as a counter adaptation to female-imposed limits on copulation. PMID- 15801588 TI - Global patterns of introduction effort and establishment success in birds. AB - Theory suggests that introduction effort (propagule size or number) should be a key determinant of establishment success for exotic species. Unfortunately, however, propagule pressure is not recorded for most introductions. Studies must therefore either use proxies whose efficacy must be largely assumed, or ignore effort altogether. The results of such studies will be flawed if effort is not distributed at random with respect to other characteristics that are predicted to influence success. We use global data for more than 600 introduction events for birds to show that introduction effort is both the strongest correlate of introduction success, and correlated with a large number of variables previously thought to influence success. Apart from effort, only habitat generalism relates to establishment success in birds. PMID- 15801589 TI - Sex-linked inheritance of hearing and song in the Belgian Waterslager canary. AB - Belgian Waterslager canaries have less sensitive hearing at high frequencies and produce songs with more energy at low frequencies than wild-type canaries. A backcross pedigree between Belgian Waterslager canaries and a domestic strain with wild-type song revealed inheritance patterns consistent with a factor of major effect located on the Z sex chromosome affecting both poor high-frequency hearing at 4 kHz and the relative energy in the spectra of the learned songs of males. Hearing thresholds at 4 kHz were significant predictors of the relative amount of song energy at 4 kHz for individual males. One hypothesis for the mechanistic basis of this correlation between hearing and song abnormalities is that a reduction in the ability to hear higher-frequency songs biases males towards learning lower-frequency songs. PMID- 15801590 TI - Parasite-mediated reduction in fecundity of mountain hares. AB - Mountain hare populations in Scotland exhibit regular 10 year fluctuations in abundance. Simple models of host-parasite population dynamics suggest that parasite-mediated reductions in host fecundity can cause a transition from stable to cyclic host population dynamics. We tested the hypothesis that parasites reduce hare fecundity by experimentally reducing parasite burdens and recording female survival, body condition and fecundity. We captured 41 adult female hares in October 2002; 22 were treated with Ivermectin to remove parasites and 19 were left untreated as controls. The treated and untreated hares were culled in May 2003 together with a second control group of nine unhandled hares. Treatment with Ivermectin significantly reduced the abundance of Trichostrongylus retortaeformis and increased-the fecundity of the hares, but had no measurable effect on body condition or over-winter survival. These results are consistent with the hypothesis that parasites may be a contributory cause of cycles in populations of mountain hares. PMID- 15801591 TI - Wide faces or large canines? The attractive versus the aggressive primate. AB - Hominids display marked body size dimorphism, suggestive of strong sexual selection, yet they lack significant sex differences in canine size that are commonly associated with intrasexual competition in primates. We resolve this paradox by examining sex differences in hominoid facial morphology. We show that chimpanzees, but not gorillas, exhibit clear sexual dimorphism in face width, over and above that expected based on sex differences in body size. We show that this facial dimorphism, expressed as an index, is negatively correlated with canine dimorphism among anthropoid primates. Our findings suggest that a lack of canine dimorphism in anthropoids is not owing to weak sexual selection, but rather is associated with strong sexual selection for broader face width. Enlarged cheek-bones are linked with attractiveness in humans, and we propose that the evolution of a broad face and loss of large canines in hominid males results from mate choice. PMID- 15801592 TI - Advantages of having a lateralized brain. AB - Brain lateralization is common among vertebrates. However, despite its implications for higher-order cognitive functions, almost no empirical evidence has been provided to show that it may confer any advantage to the functioning of the brain. Here, we show in the domestic chick (Gallus gallus domesticus) that cerebral lateralization is associated with an enhanced ability to perform two tasks simultaneously: finding food and being vigilant for predators. This finding suggests that cerebral lateralization enhances brain efficiency in cognitive tasks that demand the simultaneous but different use of both hemispheres. PMID- 15801593 TI - Long-term effects of flipper bands on penguins. AB - Changes in seabird populations, and particularly of penguins, offer a unique opportunity for investigating the impact of fisheries and climatic variations on marine resources. Such investigations often require large-scale banding to identify individual birds, but the significance of the data relies on the assumption that no bias is introduced in this type of long-term monitoring. After 5 years of using an automated system of identification of king penguins implanted with electronic tags (100 adult king penguins were implanted with a transponder tag, 50 of which were also flipper banded), we can report that banding results in later arrival at the colony for courtship in some years, lower breeding probability and lower chick production. We also found that the survival rate of unbanded, electronically tagged king penguin chicks after 2-3 years is approximately twice as large as that reported in the literature for banded chicks. PMID- 15801594 TI - Uncoupling the links between male mating tactics and female attractiveness. AB - Because not all females are equally attractive, and because mating reduces the chances of getting further copulations, males should prefer better-quality mates. In this paper, we use the Trinidadian guppy (Poecilia reticulata) to explore the effects of two non-correlated measures of female quality--size and reproductive status--on male mating decisions. All male guppies employ two alternative mating tactics. We found that large females, particularly those from a high predation site, were the target of most sneaky mating attempts. The response persisted in fish raised under standard conditions over several generations in the laboratory. In addition, non-pregnant females received more courtship displays. We conclude that males can discriminate among females and that they uncouple their mating tactics to track different axes of quality. PMID- 15801595 TI - Moa were many. AB - Until a few hundred years ago, New Zealand hosted several species of flightless ratite birds, collectively known as the moa. We have used mitochondrial sequence data for Dinornis, the largest of the moa, and new coalescent approaches to estimate its population size prior to human arrival ca. 1000-6000 years BP. We show that, as little as 1000 years ago, Dinornis numbered between 300,000 and 1.4 million and that the standing population of all moa species was ca. 3-12 million. This estimate is an order of magnitude larger than the accepted population estimate (ca. 159,000) for all moa species at the arrival of humans and suggests that moa numbers had already declined prior to human settlement. PMID- 15801596 TI - Evidence that insect herbivores are deterred by ant pheromones. AB - It is well documented that ants can protect plants against insect herbivores, but the underlying mechanisms remain almost undocumented. We propose and test the pheromone avoidance hypothesis--an indirect mechanism where insect herbivores are repelled not only by ants but also by ant pheromones. Herbivores subjected to ant predation will experience a selective advantage if they evolve mechanisms enabling them to avoid feeding within ant territories. Such a mechanism could be based on the ability to detect and evade ant pheromones. Field observations and data from the literature showed that the ant Oecophylla smaragdina distributes persistent pheromones throughout its territory. In addition, a laboratory test showed that the beetle Rhyparida wallacei, which this ant preys on, was reluctant to feed on leaves sampled within ant territories compared with leaves sampled outside territories. Thus, this study provides an example of an ant-herbivore system conforming to the pheromone avoidance hypothesis. PMID- 15801597 TI - Biogenic magnetite in the nematode caenorhabditis elegans. AB - The nematode Caenorhabditis elegans is widely used as a model system in biological research. Recently, examination of the production of heat-shock proteins in this organism in response to mobile phone-type electromagnetic field exposure produced the most robust demonstration to date of a non-thermal, deleterious biological effect. Though these results appear to be a sound demonstration of non-thermal bioeffects, to our knowledge, no mechanism has been proposed to explain them. We show, apparently for the first time, that biogenic magnetite, a ferrimagnetic iron oxide, is present in C. elegans. Its presence may have confounding effects on experiments involving electromagnetic fields as well as implications for the use of this nematode as a model system for iron biomineralization in multi-cellular organisms. PMID- 15801598 TI - Duet singing and repertoire use in threat signalling of individuals and pairs. AB - Song-type switching rate and song matching have been shown to function as territorial signals in male solo song but, to our knowledge, seem not to have been previously studied in a duetting species. We studied the plain wren (Thryothorus modestus zeledoni), to test whether duets signalled threat through song-switching rates, or through phrase type or duet type matching. Increases in the rate of song switching appear to function as an anti-habituation device rather than as a specific signal of threat. Fitting with previous results that same-sex individuals share phrase types, but pairs do not share duet types, both males and females used duets to phrase type match to playback. Pairs, however, did not duet type match in response to playback, and this suggests that within the cooperative territory defence of the duet, each sex is targeting its aggression at same-sex competitors. PMID- 15801599 TI - Directional sexual selection on chroma and within-pattern colour contrast in Labeotropheus fuelleborni. AB - Speciation via intersexual selection on male nuptial colour pattern is thought to have been a major force in promoting the explosive speciation of African haplochromine cichlids, yet there is very little direct empirical evidence of directional preferences within populations. In this study, we used objective spectrophotometry and analyses based on visual physiology to determine whether females of the Katale population of Labeotropheus fuelleborni, a Lake Malawi haplochromine, prefer males that have higher chroma and more within-pattern colour contrast. In paired male preference tests, female Katale L. fuelleborni showed increasing preferences for males with more relatively saturated colours on their flanks. They also showed increasing preferences for males with relatively higher contrast levels among flank elements. This is the first empirical evidence, to our knowledge, for male colour as a directionally sexually selected trait within a haplochromine cichlid population. PMID- 15801600 TI - Face processing in humans is compatible with a simple shape-based model of vision. AB - Understanding how the human visual system recognizes objects is one of the key challenges in neuroscience. Inspired by a large body of physiological evidence, a general class of recognition models has emerged, which is based on a hierarchical organization of visual processing, with succeeding stages being sensitive to image features of increasing complexity. However, these models appear to be incompatible with some well-known psychophysical results. Prominent among these are experiments investigating recognition impairments caused by vertical inversion of images, especially those of faces. It has been reported that faces that differ 'featurally' are much easier to distinguish when inverted than those that differ 'configurally'; a finding that is difficult to reconcile with the physiological models. Here, we show that after controlling for subjects' expectations, there is no difference between 'featurally' and 'configurally' transformed faces in terms of inversion effect. This result reinforces the plausibility of simple hierarchical models of object representation and recognition in the cortex. PMID- 15801601 TI - A survey of publication bias within evolutionary ecology. AB - Publication bias has been recognized as a problem in ecology and evolution that can undermine reviews of research results. Unfortunately, direct tests of publication bias are extremely rare. Here, we quantify a well-discussed but, to our knowledge, previously untested form of publication bias: the publication of results with and without estimates of effect size. We find that results published without effect sizes are a biased sample of those that are published. This further complicates the already difficult task of compiling quantitative literature reviews and meta-analytic studies. PMID- 15801602 TI - Population variation in lateralized eye use in the poeciliid Brachyraphis episcopi. AB - Differential use of each hemisphere of the brain for specific tasks is a widespread phenomenon that appears to have arisen in the early history of tetrapod lineage. Despite a high degree of conformity in the development of lateralization among the tetrapods, some variation exists. The mechanisms underlying this variation remain largely unresolved. We exposed fish from regions of high and low predation pressure to a series of visual experiences, including viewing an empty compartment, a novel object and a live predator. Fish from each region differed in their preferential use of each eye to view the scenes. For example, fish from high predation regions viewed a live predator by using their right eye, whereas fish from low predation sites showed no eye preference. These results suggest that the degree of lateralization varies between populations of the same species that have been exposed to different ecological/evolutionary pressures. PMID- 15801603 TI - Diclofenac poisoning is widespread in declining vulture populations across the Indian subcontinent. AB - Recent declines in the populations of three species of vultures in the Indian subcontinent are among the most rapid ever recorded in any bird species. Evidence from a previous study of one of these species, Gyps bengalensis, in the Punjab province of Pakistan, strongly implicates mortality caused by ingestion of residues of the veterinary non-steroidal anti-inflammatory drug diclofenac as the major cause of the decline. We show that a high proportion of Gyps bengalensis and G. indicus found dead or dying in a much larger area of India and Nepal also have residues of diclofenac and visceral gout, a post-mortem finding that is strongly associated with diclofenac contamination in both species. Hence, veterinary use of diclofenac is likely to have been the major cause of the rapid vulture population declines across the subcontinent. PMID- 15801604 TI - Lifetime reproduction of a forest-dwelling owl increases with age and area of forests. AB - Loss and alteration of habitats by human actions are the largest worldwide hazard to biodiversity and viability of populations. In boreal forests of Eurasia and North America the natural habitat is changing, mainly because of forestry practices and agriculture. Although there is evidence that the diversity and abundance of animal species are lower in intensively managed than in natural forests, very little is known about how the changes in habitat composition affect reproduction and survival. The best available measure of individual performance in the wild is lifetime reproductive success (LRS), the number of offspring produced during a lifetime, because it combines both survival and reproductive success to a single measure. We show that the LRS of forest-dwelling Tengmalm's owls (Aegolius funereus) increases with the proportion of old forest in the territory because of a higher number of breeding attempts, whereas it decreases with the proportion of agricultural land because of declining fledging success in years when prey populations crashed during owl breeding. These unique results provide an interesting insight into how human influence on the landscape can affect life-history traits of animals through various pathways. PMID- 15801605 TI - How brachiopods get covered with nanometric silicon chips. AB - The investigation of an early pelagic juvenile of the discinid brachiopod Discinisca cf. tenuis elucidates the so far enigmatic origin of nanometric silicon chips covering the brachiopod's juvenile shell. The siliceous tablets are products of an intracellular process within specialized cells of the animal's inner mantle epithelium. These specialized cells are arranged in a circumferential row and contain vesicles, which provide 'reaction chambers' osmotically separated from the cytoplasm. Up to 15 tablets per vesicle are released into the cell by vesicle burst, followed by a coordinated extrusion onto the periostracum. In conjunction with the conveyor belt mechanism of periostracum formation, the regime of tablet release accounts for the highly ordered arrangement of siliceous tablets in parts of the shell's surface. The siliceous tablets are discussed as a protective cover against solar radiation, inherited from Palaeozoic linguliform brachiopods. PMID- 15801606 TI - Contagious yawning in chimpanzees. AB - Six adult female chimpanzees were shown video scenes of chimpanzees repeatedly yawning or of chimpanzees showing open-mouth facial expressions that were not yawns. Two out of the six females showed significantly higher frequencies of yawning in response to yawn videos; no chimpanzees showed the inverse. Three infant chimpanzees that accompanied their mothers did not yawn at all. These data are highly reminiscent of the contagious yawning effects reported for humans. Contagious yawning is thought to be based on the capacity for empathy. Contagious yawning in chimpanzees provides further evidence that these apes may possess advanced self-awareness and empathic abilities. PMID- 15801607 TI - The effects of a severe drought on mouflon lamb survival. AB - The mouflon population of Caroux-Espinouse, southern France, inhabits a highly seasonal area with dry summers. We monitored summer lamb survival during a severe drought in 2003, from early June to late August. The survival of 35 radio-tagged lambs over nine two-week periods was strongly affected by the timing of rainfall. Survival depended on the amount of rainfall recorded at a given 14 day period and in the previous 14-21 day period. Survival was not influenced by the exceptionally high mean daily temperature recorded during some periods. Male lamb survival (0.68) tended to be less than female survival (0.81), although not significantly, possibly because of a low sample size. The high lamb mortality (25.7%) recorded during a four-month period is much higher than previous estimates of first-year mortality (less than 10%). We recommend accounting for climatic variation in summer when studying the population dynamics of ungulates. PMID- 15801608 TI - Subjective life expectancy predicts offspring sex in a contemporary British population. AB - There is evidence that women who are in poor physical condition or who reside in deprived environments are more likely to give birth to daughters than to sons. Under deprived environmental conditions, or when in poor physical health, it has been hypothesized that parents should take into account the available resources and manipulate the sex of any children born. Using subjective life expectancy (SLE) as a measure of how an individual views their future health and environment, I demonstrate that there is an association between the sex of the first child and SLE in a sample of mothers from a contemporary British population (Gloucestershire, UK). SLE was a significant predictor of offspring sex: women who believed that they had longer to live were more likely to have had a male birth than women who thought they would die earlier. Detection of such a bias among the children of British mothers may provide evidence that the sex ratio under relatively affluent Western conditions can still be influenced by adverse environmental or poor maternal condition. PMID- 15801609 TI - Identifying the role that animals play in their social networks. AB - Techniques recently developed for the analysis of human social networks are applied to the social network of bottlenose dolphins living in Doubtful Sound, New Zealand. We identify communities and subcommunities within the dolphin population and present evidence that sex- and age-related homophily play a role in the formation of clusters of preferred companionship. We also identify brokers who act as links between sub-communities and who appear to be crucial to the social cohesion of the population as a whole. The network is found to be similar to human social networks in some respects but different in some others, such as the level of assortative mixing by degree within the population. This difference elucidates some of the means by which the network forms and evolves. PMID- 15801610 TI - Evidence for 'cross-talk' between A and B chromosomes of rye. AB - Spontaneous DNA insertions from supernumerary B chromosomes (Bs) into the standard A chromosome complement were detected in rye (Secale cereale L.), using fluorescent in situ hybridization (FISH) analysis with the D1100 B-specific sequence probe. The insertions were seen in individuals derived from plants possessing deleted Bs, characterized in this study by not having the B-specific sequences that are normally found at the distal part of the long arm of the standard rye B. This result supports the case for the spontaneous introgression of B-specific DNA into the A chromosome genome, and it indicates that 'cross talking' between A and B chromosomes may occur in wild populations. PMID- 15801611 TI - Nocturnal colour vision in geckos. AB - Nocturnal animals are said to sacrifice colour vision in favour of increased absolute sensitivity. This is true for most vertebrates that possess a dual retina with a single type of rod for colour-blind night vision and multiple types of cone for diurnal colour vision. However, among the nocturnal vertebrates, geckos are unusual because they have no rods but three cone types. Here, we show that geckos use their cones for colour vision in dim light. Two specimens of the nocturnal helmet gecko Tarentola (formerly Geckonia) chazaliae were able to discriminate blue from grey patterns by colour alone. Experiments were performed at 0.002 cd m(-2), a light intensity similar to dim moonlight. We conclude that nocturnal geckos can use cone-based colour vision at very dim light levels when humans rely on colour-blind rod vision. PMID- 15801612 TI - The social implications of winner and loser effects. AB - Winner and loser effects have now been documented in a number of species. To our knowledge, experimental work, however, has focused exclusively on pairwise interactions, and not the extent to which winner and loser effects impact hierarchy formation. We report the results of experimentally manipulated winner and loser effects on hierarchy formation in a socially living species, the green swordtail, Xiphophorus helleri. Our results demonstrate that randomly chosen winners in pairwise aggressive contests were more likely to emerge as top-ranked individuals in a hierarchy, whereas randomly chosen losers were more likely to emerge as the lowest-ranking individuals, and that 'winner-neutral-loser' hierarchies were significantly overrepresented. PMID- 15801613 TI - Evidence for the impact of global warming on the long-term population dynamics of common birds. AB - Taking the opportunity in 2003 of the exceptionally warm spring in France as a natural simulation of possible future climate, we analysed common bird productivity using the French long-term capture-recapture national monitoring scheme. Two-thirds of the 32 species studied had an above-average productivity in 2003. However, this gain in productivity was not consistent among species, with a relatively low productivity for species exhibiting a long-term decline and relatively high productivity for stable or increasing species. Such links between long-term and short-term dynamics suggest that the impact of increasingly warm springs on productivity is a major component of the recent population dynamics of a variety of common bird species. PMID- 15801614 TI - 'Junk' DNA and long-term phenotypic evolution in Silene section Elisanthe (Caryophyllaceae). AB - Nuclear DNA content variation over orders of magnitude across species has been attributed to 'junk' repetitive DNA with limited adaptive significance. By contrast, our previous work on Silene latifolia showed that DNA content is negatively correlated with flower size, a character of clear adaptive relevance. The present paper explores this relationship in a broader phylogenetic context to investigate the long-term evolutionary impacts of DNA content variation. The relationship between nuclear DNA content and phenotype variation was determined for four closely related species of Silene section Elisanthe (Caryophyllaceae). In addition to a consistent sexual dimorphism in DNA content across all of the species, we found DNA content variation among populations within, as well as among, species. We also found a general trend towards a negative correlation between DNA content and flower and leaf size over all four species, within males and females as well as overall. These results indicate that repetitive DNA may play a role in long-term phenotypic evolution. PMID- 15801615 TI - Androgens, interspecific competition and species replacement in hybridizing warblers. AB - The steroid hormone testosterone regulates aggressive behaviour in many vertebrates and is important for territorial defence among males of the same species. However, its role in mediating interspecific competition, and ultimately species distributions, is unknown. We show that testosterone may influence the geographical replacement of one species by another. Townsend's warblers (Dendroica townsendi) have replaced hermit warblers (D. occidentalis) over a vast portion of their historical range, partly because Townsend's males are more aggressive than hermit males and outcompete them for territories in areas of sympatry. We report differences in plasma androgen levels that parallel these aggressive asymmetries and the historical pattern of species replacement between Townsend's and hermits. Using hybrids, we provide evidence that these hormonal differences are partially genetically based and thus may have evolved through sexual selection during Pleistocene glacial maxima. Hormone-behaviour mechanisms can therefore have important effects on species distributions and can even influence the pathways underlying extinction. PMID- 15801616 TI - Generation cycles in Indonesian lady beetle populations may occur as a result of cannibalism. AB - Generation cycles, population cycles with a period of approximately one generation, have been observed in a variety of field and laboratory studies. Such dynamics are predicted to arise through the effects of resource competition and cannibalism or involve consumer-natural enemy interactions. We first show, using a new highly simplified model, that generation cycles are a very common outcome of strongly age-structured intraspecific interactions involving cannibalism. We then analyse a series of unique long-term time-series of ladybeetle (Coccinellidae) abundances from tropical Indonesia. Some of the time-series display clear generation cycles, and we argue that there is strong evidence that these are caused by intraspecific cannibalism. PMID- 15801617 TI - Strategic growth decisions in helper cichlids. AB - Recently, it has been shown that group-living subordinate clownfish Amphiprion percula increase their growth rate after acquiring the dominant breeder male position in the group. Evidence was found for strategic growth adjustments of subordinate fishes depending on the threat of eviction, i.e. subordinates adjust their growth rates so they remain smaller than the dominant fish and thereby limit the threat of being expelled from the territory. However, it is impossible to exclude several alternative factors that potentially could have influenced the observed changes in growth, owing to the nature of that experiment (removing the second-ranking fish--the breeder male--caused the third-ranking fish to change sex to become breeder male and change rank). We studied strategic growth decisions in the group-living Lake Tanganyika cichlid Neolamprologus pulcher under controlled laboratory conditions with ad libitum food availability. First, we show that male breeders grow faster than subordinate male helpers of the same initial size and confirm that N. pulcher shows status-dependent growth. Second, we improved on the experimental design by not removing the dominant breeder male in the group; instead we replaced the breeder male with a new breeder male in a full factorial design and measured growth of the subordinate male helpers is a function of the size difference with the old and the new breeder male. As predicted, male helpers showed strategic growth adjustments, i.e. growing faster when the size difference with the breeder male is large. Strategic growth adjustments were less pronounced than status-dependent growth adjustments. PMID- 15801618 TI - Queen and worker policing in monogynous and monandrous colonies of a primitively eusocial wasp. AB - Kin-selection theory predicts that a worker prefers to produce her own sons in a colony with monandry and monogyny because relatedness to her sons (0.5) and nephews (0.375) exceeds that to brothers (0.25). In spite of this prediction, recent studies reveal that workers police each other (mutual-worker egg removal) even in monandrous and monogynous colonies. We conducted field and laboratory studies to evaluate queen and worker policing in queen-right colonies of the primitively eusocial wasp Polistes chinensis antennalis. Genetic studies using microsatellite markers, as well as extensive observations of natural colonies, revealed that both queen and workers removed both queen- and worker-laid eggs in monogynous and monandrous colonies. The queen's eggs survived to hatching more successfully than those of the workers (88.5% versus 1.4%). We discuss the likely factors to explain these worker-policing behaviours. PMID- 15801619 TI - Distress calls may honestly signal bird quality to predators. AB - In predator-prey interactions, both interactors may benefit from sharing information about prey vulnerability. We examined the relationship between calls used to discourage close predators (distress calls) and the health condition of the caller to test whether these signals are reliable indicators of prey quality. The structure of calls from captured lesser short-toed larks Calandrella rufescens was related to their body condition and T-cell-mediated immunocompetence. Birds in better nutritional and immunological condition utter harsher calls (i.e. they spread the call energy over a wider range of frequency) than birds in poorer conditions. Hence, the harshness of distress calls seems honestly to signal the health status of prey and thus their ability to escape, on which the predator might base its optimal foraging choice. Previous studies have investigated the honesty of songs that have evolved via sexual selection, but this is the first study, to our knowledge, the demonstrates a relationship between individual quality and a vocalization primarily shaped by natural selection. PMID- 15801620 TI - Social networks in the guppy (Poecilia reticulata). AB - Social network theory is used to elicit details of the social structure of a population of free-ranging guppies, Poecilia reticulata. They were found to have a complex and highly structured social network, which exhibited characteristics consistent with the 'small world' phenomenon. Stable partner associations between individuals were observed, a finding that fulfils the basic prerequisite for the evolution of reciprocal altruism. The findings are discussed in relation to the ecology and evolution of the wild population, highlighting the potential application of network theory to social associations in animals. PMID- 15801621 TI - ["Veterinary activities": the comeback]. PMID- 15801622 TI - [First documentation of a neospora-induced "abortion storm" (exogenous transplacental transmission of neospora caninum) in a Swiss dairy farm]. AB - In a Swiss dairy farm (canton of Geneva) consisting of 73 animals 8 abortions were observed within 2 weeks. Serological and molecular biological analyses (PCR) on aborting dams, and abortion materials, respectively, revealed that the protozoan parasite Neospora caninum was the causative agent. Besides the 8 aborting animals, 12 other non-aborting heifers were found to be serologically positive for this parasite. All positive sera were further tested in an avidity ELISA to elucidate the recency of infection. All seropositive animals but one showed low avidities at the time the abortion storm started. This indicated at a recent N. caninum-infection within the herd. Thus, the animals most probably were exposed to N. caninum-oocysts (e.g. by dog feces-contaminated forage) and the resulting abortion storm was due to an exogenous (formerly known as "horizontal") parasite transmission into a naive herd. This is the first documented record of such an event in Switzerland. PMID- 15801623 TI - [Autochthonous infections with Angiostrongylus vasorum in dogs in Switzerland and Germany]. AB - Angiostrongylus vasorum is endemic in foxes an other carnivores in the South and south East of France, Denmark and Great Britain. The reddish nematode is present in the Arteria pulmonalis and the right side of the heart and causes respiratory and cardiovascular symptoms. From 1999 to 2004, A. vasorum was diagnosed in 5 dogs from northern Switzerland, in 1 dog from southern Germany and in 3 dogs from south Ticino. Clinical signs in the affected dogs varied and ranged from cough, tachypnoea and dyspnoea to neurological symptoms in 2 of the dogs. Four dogs died and in 3 of the 4 dogs adults and larvae were found after digestion of formalin fixed tissue. Diagnosis in the other 5 dogs was achieved by detecting the larvae coproscopically. Based on anamnestic data, these 9 dogs are probably autochthonous cases. PMID- 15801624 TI - Clinicopathologic observations on Coenurus cerebralis in naturally infected sheep. AB - Twelve sheep from 7 different flocks consisting of approximately 150-250 animals each were diagnosed with coenurosis caused by the larval stage of Taenia multiceps. Ataxia, incoordination, drowsiness, hind leg paralysis and coma were the most prominent clinical symptoms. Monocytosis and lymphocytosis were observed upon hematological examination. Creatin kinase (CKBB) levels of the animals varied between 421 and 495 U/l. Cysts were commonly localized in the parietal and frontal lobes of the brain and in the cerebellum. In two cases, cysts were found on the lumbar aspect of the medulla spinalis. Symptoms were related to cyst localization. Depression, tilting of the head either to the right or left and head pressing were seen when cysts were located in the cerebrum. Incoordination and hyperexcitability were noted if the cysts were involved with the cerebellum and when located in the spinal cord, hind leg paralysis was the typical clinical sign. On microscopic examination, atrophy was observed in the central nervous system (CNS) organs due to pression by the bladderworms. Nonpurulent meningoencephalitis with perivascular cuffings were the most common histopathological findings. In periodic acid Schiff staining (PAS), positive reaction was observed in protoscoleces. Neurons were the most affected cell type when stained by the Kluver Barrera method. This method also showed that in the CNS, Coenurus cerebralis caused a prominent glial reaction. When parasites were localized in the nervous system treatment was impossible. Animals without neurologic sings were treated with praziquantel (Tenikur tablet-Topkim A.S.) 50 100 mg/kg/day for three days. PMID- 15801625 TI - [Phytomedicine in dogs and cats: web-based survey among veterinarians in Austria, Germany and Switzerland]. AB - In the last decades phytopharmaceuticals have stabilized their significance in human medicine. The aim of this study was to obtain information about the present situation of phytomedicine in three German-speaking countries. For that purpose questionnaires were sent via internet to 2675 veterinarians in Austria, Germany and Switzerland and the collected 189 answers were statistically evaluated. As shown in this paper, there is an increasing interest in veterinary phytotherapy. Presently three-quarters of the veterinarians are using phytoproducts in clinical practice. Herbal medicinal products are only seldom used in eye and ear infections as well as in endo- and ectoparasitoses but more often in diseases of the liver, joints, kidneys, heart and skin. Generally, phytodrugs are more frequently applied in chronic diseases as well as secondary medication, whereas the time of treatment usually exceeds one month. Basic advantages of medicinal phytoproducts have been shown to be the low side effects and the acceptance of the pet-owners and the only disadventage has been reported to be the lack of information. This study clearly shows the upward trend in using herbal drugs by veterinarians but the results also point to the lack of scientific knowledge about phytotherapeuticals in animal health. PMID- 15801626 TI - [Criteria for assessing antituberculosis care]. PMID- 15801627 TI - [Major risk factors of tuberculosis in children and adolescents]. PMID- 15801628 TI - [Detection of tuberculosis in children and adolescents in the Primorye Territory]. AB - The epidemiological indices and the methods for detecting tuberculosis have been analyzed in the children and adolescents of the Primorye Territory. It has been established that morbidity among children became stable by 2003 and was 23.6 per 100,000; in adolescents it remains high - 79.5 per 100,000 in 2003. The association of the role of different detection methods with the age group is now a specific feature of tuberculosis in children and adolescents. Mass tuberculin diagnosis plays a decisive role in the detection of tuberculosis only in children aged 5-7 years. In other age groups, tuberculosis is frequently identified at contacts with a physician. Radiation is the leading technique in detecting tuberculosis in adolescents. The consideration of the specific features of tuberculosis detection in relation to the age group assisted one in the selecting the most optimum strategy of tuberculosis control in children and adolescents. PMID- 15801629 TI - [Tuberculosis in children from the Sverdlovsk Region]. AB - The tuberculosis morbidity rate in BGC-unvaccinated infants is 18 times greater than that in BCG-vaccinated infants and 6 times higher than the rate of complications due to BCG vaccination. If a scar due to BCG vaccination is absent or 1-2 mm, the tuberculosis morbidity in children of 3 years of life is 7 times higher than when a 3-mm or more scar has formed after vaccination. The technological failures are main causes of complications due to BCG vaccination. The type of a vaccine (BCG-1 or BCG-M) does not play a substantial role in the development of complications. PMID- 15801630 TI - [Significance of a children's tuberculosis sanatorium]. AB - The paper analyzes a contingent of children and adolescents from a tuberculous sanatorium in 2000-2002. It was established that the children and adolescents with local forms of tuberculosis amounted to 14.5%, the children with a variety of tuberculin reactions were 33.5%; the children infected with tuberculosis and risk factors were 42.1%; the total proportion of children who had contacted patients with tuberculosis was 31.7% of the whole contingent of the sanatorium, i.e. about a third. In addition to different manifestations of tuberculous infection, nonspecific abnormalities were detected in 96.4% of the children and adolescents, in the vast majority they being first diagnosed at the sanatorium. The foregoing suggests that a children's tuberculosis sanatorium is of great medical importance under the present conditions. PMID- 15801631 TI - [Ways of optimizing the work of tuberculosis sanatoriums for children and adolescents]. AB - The activities of two federal and regional tuberculosis sanatoriums for children and adolescents were analyzed. The clinical characteristics and the results of the examination and treatment were studied in 510 patients aged 12-17 years who had various manifestations of tuberculous infection. The beds in the sanatoriums were ascertained to occupy irrationally: the proportion of patients with local forms of tuberculosis there is not more than 30%. Guidelines for selection of children and adolescents with tuberculosis from risk groups and for detection of the disease were developed. A proposal was made to set up consultative and diagnostic centers at the sanatorium basis. The paper shows it possible and expedient to treat children and adolescents with new-onset local, non-destructive forms of tuberculosis under the conditions of a sanatorium. PMID- 15801632 TI - [The reserve drugs: their safety for children and adolescents]. AB - For the treatment of children and adolescents with tuberculosis, the needs for reserve drugs are 24.0 and 48.6%, respectively. The main reason of the use of reserve drugs is intolerability of essential antituberculous drugs in children (16.4%), and resistance to these drugs in adolescents (40%). Overall, different types of intolerability events (allergic, toxic, toxicoallergic reactions) were observed in 27 (34%) of 79 children and in 46 (65%) of 70 adolescents. Adverse reactions to essential drugs were significantly more frequently observed than those to reserve drugs in both the children and the adolescents: in 96.3 and 3.7% (p < 0.001) and in 89.1 and 10.9% (p < 0.001) cases, respectively. PMID- 15801633 TI - [Prophylaxis of tuberculosis with BCG vaccine in children from contact and from unknown contact]. AB - The quality and efficiency of immunoprophylaxis against tuberculosis have been studied in 563 children, by estimating the vaccination BCG signs. The children who have familial contacts and have been rather fully (92.3%) and effectively inoculated with BCG vaccination and in whom skin scars, 4-7 mm in diameter, have formed are at a high risk of tuberculosis due to massive infection in the bacterial isolation foci. Inadequate immunobiological defense reactions promote the progression of endogenous infection and the development of active tuberculosis in the vast majority (67.8%) children who have unestablished and unvaccinated or ineffectively vaccinated with BCG, infected under the occult bacterial isolation conditions mainly in the families where the adults have residual pulmonary changes and hypersensitivity to tuberculin the absence of immunobiological defensive reactions is the major risk factor for active tuberculosis. PMID- 15801634 TI - [An inflammation reaction in the postvaccinal keloid scar]. PMID- 15801635 TI - [A role of individual tuberculin diagnosis in the formation of registry group VI]. AB - The authors studied the use of comprehensive individual tuberculin diagnosis (scarification skin test, intradermal titer) in the differential diagnosis of the nature of tuberculin sensitivity in children referred for examinations by physiological pediatricians for the diagnosis of a turn of tuberculin tests and the formation of registry group VIA. The use of this procedure was found to avoid the hyperdiagnosis of a turn of tuberculin tests in as high as 55.8%, to improve the diagnosis of hyperergic reactions to tuberculin, and to detect an occult false hyperergic reaction. Moreover, there is a possibility of observing the changes in individual sensitivity in Registry Group 0 children, which contributes to their more timely transfer to Group VIA. Analyzing the results shows that the use of comprehensive tuberculin diagnosis promotes the optimization of a system of preventive measures aimed at preventing tuberculosis in children. PMID- 15801636 TI - [Intubation tube-induced tracheal ruptures]. PMID- 15801637 TI - [Pulmonary hemorrhages of different etiology: diagnosis and treatment]. AB - Hemorrhage is one of the most pressing problems in lung diseases. The paper analyzes the results of medical and surgical treatments for pulmonary hemorrhages. A total of 724 patients with hemoptysis and bleedings of various etiology were treated at the Thoracic Unit in 1985-2003. Pulmonary hemorrhages occurred in 442 (61%) patients with pulmonary tuberculosis and in 282 (39%) patients with nonspecific lung diseases. X-ray study revealed right-sided lung lesions in 268 (37%) patients; left-sided lung lesions in 203 (28%), bilateral lesions in 170 (23.5%). The visible X-ray changes were not found in 83 (11.5%) patients. Destructive pulmonary parenchymal changes were detected in 366 (50.5%) patients. Bronchological study was conducted in 703 patients with pulmonary hemorrhage: 377 (53.7%) and 214 (30.4%) patients were found to have its direct and indirect signs, respectively. Neither changes nor hemorrhagic traces were detected in 112 (15.9%) patients since they underwent bronchoscopy in a later period when bleeding had already stopped. Foam rubber sponge tamponade of the main and lobar bronchi was made in 75 (10.4%) patients with profuse bleeding; a positive effect was achieved in 61 (81.3%) patients. The treatment policy for pulmonary hemorrhage is determined by its severity and etiology and the health status of the patient. A major role was assigned to medical hemostatic therapy in the treatment of pulmonary hemorrhages. If it was ineffective, surgical treatment was used. The use of hypotension controlled by ganglionic blockers (more commonly pentamine): bleedings were suppressed in 359 (74%) of 485 patients with second- and third-degree hemorrhages yielded good results. They were medically stopped in 558 (77%) patients; death occurred in 14 (2.4%) cases. A hundred and fifty two (20.9%) were operated on; of them 8 patients underwent segmentectomy; 85 had lobectomy. Pulmonectomy, cavernostomy, and thoracoplasty were performed in 48, 4, and 7 patients, respectively. Twenty-six patients were operated on at the peak of hemorrhage. Bleedings were suppressed in 140 (92%) patients (they recovered; 12 (7.9%) patients died. In the postoperative period different complications were noted in 30 (19.7%) patients. The final results of medical and surgical treatment were as follows: bleedings were stopped in 698 (96.4%) patients (they recovered) and 26 (3.6%) died. PMID- 15801638 TI - [Risk factors for early relapse of tuberculosis in Latvia]. AB - OBJECTIVE: To evaluate risk factors for tuberculosis relapse within a short time after successful tuberculosis treatment among Latvian patients treated in 1996. DESIGN: Retrospective case-control study of tuberculosis patients diagnosed and treated successfully in 1996. RESULTS: 48 cases and 96 matched controls were included in the study. 18 cases and 22 controls had documentation of at least one interruption (OR 2.96; 95% CI 1.09-7.99). Early relapse was not associated with type of treatment facility, age or unemployment, but was positively associated with alcohol use (OR 17.70; 95% CI 4.14-75.77). In a multivariate analysis, only alcohol use was significantly associated with early relapse (OR 16.63; 95% CI 3.63-76.10). CONCLUSION: Alcohol use was the main risk factor for early tuberculosis relapse. PMID- 15801639 TI - [The satisfaction of personal needs in patients with pulmonary tuberculosis]. AB - The founder of humanistic psychology A. Maslow divided the needs of a personality into several levels--from the lowest to the highest ones. Higher-leveled needs rise when the lower-leveled needs are satisfied. A great deal of factors affect the origination and satisfaction of needs, but they are always interrelated with social values. The extent to which personality needs are satisfied in patients with pulmonary tuberculosis has not been studied. A special questionnaire has been drawn up to study the extent to which personality needs are met. Its suitability has been determined, by using a group of patients with bronchial asthma. The extent to which personality needs are satisfied in patients with pulmonary tuberculosis was studied in 178 patients with infiltrative pulmonary tuberculosis and 253 patients with fibrocavernous pulmonary tuberculosis. The results have shown that the extent to which personality needs are satisfied in patients with tuberculosis is lower than that in apparently healthy individuals of the same social status. In females with infiltrative pulmonary tuberculosis, the needs for safety are satisfied to a lesser extent. In those with fibrocavernous pulmonary tuberculosis, the extent to which the physiological, safety, and self-realization needs is decreased. In males with infiltrative pulmonary tuberculosis, the physiological, noetic, and self-realization needs are satisfied to a lesser extent. In those with fibrocavernous pulmonary tuberculosis, the extent to which the physiological, safety, group decision making, noetic and self-realization needs is decreased. PMID- 15801640 TI - [Assessment of postload glycemic curves in the prediction of carbohydrate metabolic disturbances in patients with bronchial asthma]. AB - The sensitivity and responses to exogenous insulin were studied in 78 patients with bronchial asthma. The study identified 6 types of postinsulin blood glucose concentration curves that reflected hormonal control impairments of different phases. Decreased insulin sensitivity, a delayed biological effect, and insulin resistance may serve as the early predictors of carbohydrate metabolic disturbances and severe metabolic pathology. PMID- 15801641 TI - [Zakharin as a phthisiologist]. PMID- 15801642 TI - [Laser therapy in the combined treatment of pulmonary tuberculosis in adolescents (a guide for the physician)]. PMID- 15801643 TI - [Impact of consultation on erectile dysfunction in the current and future urology practice]. PMID- 15801644 TI - [Surgical treatment of female urethral diverticula. Our experience. Bibliographic review]. AB - OBJECTIVES: Female urethral divertilum is a rare disease; its diagnosis has increased with the use of new diagnostic technologies. It must be suspected in women consulting for chronic irritative symptoms without response to conventional treatments. Transvaginal surgical excision is the most accepted therapeutic option. We reviewed their clinical presentations, diagnostic findings, and therapeutic options and report our experience. METHODS/RESULTS: We retrospectively reviewed our case series, finding 4 patients with the diagnosis of female urethral diverticulum; we performed a bibliographic review. CONCLUSIONS: Urethral diverticulum is a rare clinical entity which has to be included in the differential diagnosis of women with chronic lower urinary tract symptoms. Clinical presentation may vary from asymptomatic to rich voiding symptoms. The most frequently used diagnostic method is voiding cystourethrogram; other techniques such as transvaginal ultrasound or MRI are very useful for complicated cases. Surgical treatment by transvaginal diverticulectomy with closure in several layers is the most frequent approach. Postoperative complications are rare. PMID- 15801645 TI - [Multilocular cystic renal carcinoma: pathologic study of 14 cases]. AB - OBJECTIVES: The article focuses on the multilocular cystic renal carcinoma, the most frequent of the renal cystic carcinomas. The importance of performing a separate analysis of these cystic tumors comes from their better prognosis some authors have suggested. Moreover, they are frequently difficult to differentiate from multilocular benign cysts and other benign lesions by radiological studies, cytology or even during surgery. Pathologic study is essential for a definitive diagnosis of the lesion. METHODS AND RESULTS: We reviewed 14 cases of multilocular cystic renal carcinoma obtained from the series of 191 renal cell carcinomas diagnosed in our hospital in the period 1995-2002. Inclusion criteria were existence of a multicystic complex mass, in which clear cells partially or completely covered the cystic walls and accumulate within the septa. CONCLUSIONS: The lack of well-documented cases of multilocular cystic renal carcinoma with metastases supports the fact that they are tumors with a very good prognosis. In conclusion, data from both literature and our series support that multilocular cystic renal carcinoma should be considered a low-grade neoplasia, and its treatment should be conservative. PMID- 15801646 TI - [Local recurrence after radical prostatectomy and salvage radiotherapy]. AB - OBJECTIVE: To evaluate the efficacy of radiotherapy to the prostatic bed in patients with biochemical recurrence prostate cancer after radical prostatectomy. METHODS: We analyse the outcomes of 292 patients who underwent radical prostatectomy for localized prostate cancer T1-T2 between January 1992 and June 2003, with an average follow-up of 36 months (range 6 months to 12 years). We detected biochemical recurrence (PSA > 0.20 ng/ml) in 75 (26%) patients. 75 patients with biochemical recurrence, 9 (12%) were diagnosed of local recurrence by the following criteria: a) First PSA obtained 6 weeks after radical prostatectomy < 0.20 ng/ml. b) Time to biochemical recurrence > 6 months. c) Prostate specific antigen doubling time > 6 months. d) Prostate specific antigen velocity after radical prostatectomy < 0.75 ng/ml/year. e) Prostate specific antigen level after radical prostatectomy < 2.5 ng/ml. The 9 patients diagnosed of local recurrence received an average dose of 56.42 Gy to the prostate bed. RESULTS: Of all 9 patients with local recurrence, 8(88.8%) have complete response with a mean follow-up of 30 months (12-36 months). The time between the radiotherapy and the response, in patients with complete response, was lower than 3 months in 7 patients and 12 months in 1 patient. Significant adverse effects associated to radiotherapy were not observed. CONCLUSIONS: Salvage radiotherapy may be beneficial in selected patients with local recurrence. The characteristics of prostate specific antigen elevation are useful in distinguishing men with local recurrence from those with distant metastases. PMID- 15801647 TI - [Urodynamic paradigms: from renaissance fibrilar theory to current Doppler flowmetry]. AB - To perform a historical introduction and a review of the mathematical model, emphasizing that our mathematical model may be the solution to the viscoelastic model. It is evident that the same experiment has been repeated over half a century, with similar results in all cases. We also show one of the projects we are working on: the electro-vesicogram for the evaluation of the filling phase, and Doppler uroflowmetry for the study of the voiding phase. METHODS: We have chosen and studied in depth the results Dr. Virseda presents in his thesis of one of the experiments performed in relation to the viscoelastic model. After applying analytical methods we reach a differential equation we suppose defines detrusor behaviour, as it has been explained by the viscoelastic model. The solution of this equation by means of the Laplace's transform enables to obtain the values of the incognitas set by urodynamics. Besides, we analyzed the behaviour of solutions' stability using a matricial method following the Lyapunov theory. The former may solve the incognitas for the voiding phase. We used urethral Doppler with simultaneous uroflowmetry to obtain the data equations demanded; this is what we named "Doppler uroflowmetry". The filling phase was studied by superficial electromiography. We named it "electrovesicogram". We attach images for both Doppler wave and electrovesicogram. They both are the projects we are working on. RESULTS: Currently we can only explain the methodology we are following. Indeed, this article is the first of a series in which we aim to explain the methodology we are following in detail: Doppler wave capture; mounting process photogram by photogram, and vectorization and cleaning of the wave, either Doppler or flow waves; treatment in autocad to obtain the vector; and management of the vector with the matalab software, which gives us the results we are looking for. CONCLUSIONS: It is intuitive to deduct the usefulness of these methods as not invasive techniques in the urodynamic diagnosis. We have our illusions in these projects which open a window to the future. PMID- 15801648 TI - [Impotence and cardiovascular disease: a new perspective in the health-care compromise of the urologist]. AB - OBJECTIVES: To review the evidence currently available to establish the pathogenic connection between erectile dysfunction (ED) and cardiovascular disease (CVD), and the effects this scientific progress has introduced in the classical impotence urology clinic. METHODS: We reviewed the most recent publications about this disease (2000-2004) and the concept of endothelial dysfunction by appropriate MEDLINE searches, with specific selection of reviews and clinical practice guidelines. RESULTS: The fact that ED and CVD share risk factors is confirmed; the pathogenic unity of both processes having endothelial dysfunction as the underlying problem; anticipation of ED over CVD in time of presentation; notable increase of research about this issue over the last two years; the change of scenario in the impotence urology clinics due to these findings. CONCLUSIONS: The number of cases in which ED is not an organ disease but an early symptom of endothelial dysfunction forces changes in the extent and depth of the diagnostic, prognostic and follow-up strategies in the urology impotence clinics of extraordinary importance from both the individual and health care politics point of view. PMID- 15801649 TI - ["Mini-percutaneous" percutaneous nephrolithotomy: technique and indications]. AB - OBJECTIVES: Since Helal and Jackman developed the mini-percutaneous ("mini-perc") percutaneous nephrolithotomy (PCNL) for the treatment of pediatric renal lithiasis various authors have used is technique in adults with the aim to preserve renal parenchyma and diminish morbidity associated with the standard PCNL. METHODS: We describe the surgical technique of "mini-perc" PCNL in the treatment of renal lithiasis. RESULTS: We review various case series from the literature of "mini perc" PCNL for the treatment of adult renal lithiasis, its current indications, percentages of stone free patients, and associated problems, mainly the required instruments. CONCLUSIONS: Morbidity associated with "miniperc" PCNL seems to be lower than with the standard PCNL, but the use a smaller calibre tract with the only aim to preserve renal parenchyma does not offer advantages. It is a complementary technique to the standard PCNL; however, the diameter of the percutaneous access is not well defined yet and depends on the calibre of the instruments used. PMID- 15801650 TI - [Malignant recurrence of a penile cutaneous horn]. AB - OBJECTIVES: To report one case of malignant recurrence of a cutaneous horn of the glans penis. METHODS: We report the case of a 66-year-old male patient presenting with penile discomfort over several months. Physical examination showed a corneal lesion in the glans-penis, the biopsy of which was reported as hyperkeratosis on top of pseudoepiteliomatous hyperplasia. Partial penectomy of the glans penis was performed after recurrence three months later. RESULTS: Pathological study of the specimen identified a microinvasive squamous cell carcinoma in the base of the lesion. The patient is disease-free after two years of follow-up. CONCLUSIONS: We emphasize the capacity the penile cutaneous horn has to become malignant. We also point out the need for observation of these lesions after excision. PMID- 15801651 TI - [Renal malacoplakia: report of a new case]. AB - OBJECTIVES: Malacoplakia is a chronic granulomatous disease which can involve different areas of the body, being the genital renal system the most frequent site. We describe a case of unilateral renal malacoplakia. METHODS/RESULTS: We report the case of a female patient with the clinical working diagnosis of renal cell carcinoma that required drainage of a homolateral abscess which cultured positive for E. Coli. Nephrectomy was performed. Pathologic study showed the presence of Michaelis-Gutmann bodies, patognomonic of malacoplakia. PMID- 15801652 TI - [Breast metastasis as initial presentation of a renal carcinoma]. AB - OBJECTIVES: To report the case of a breast metastasis as initial presentation of renal carcinoma. METHODS: 72-year-old male patient who consulted for a painful right breast tumor. We describe clinical history, complementary tests, biopsy and treatment. RESULTS: Pathologic study confirmed a metastasis of a renal clear cell carcinoma. A CT scan confirmed the existence of bilateral renal tumors. CONCLUSIONS: Breast metastases are exceptional as initial presentation of a renal carcinoma. We performed a bibliographic review on the topic. PMID- 15801653 TI - [Cognitive and behavioural alterations as adverse reaction to tamsulosin not previously described]. AB - OBJECTIVES: To report one case of adverse reaction to tamsulosin. METHODS: We describe the case of an 80-year-old patient who was started on alpha blocker treatment after acute urinary retention with the aim to perform a voiding test. RESULTS: The patient presented self-limited episodes of disorientation in time and space and abnormal behaviour in coincidence with the first three administrations of the drug. Treatment was stopped. CONCLUSIONS: We report one case of an adverse reaction to tamsulosin not previously described. We make a reference to the drug vigilance mechanisms. We point out the importance of coordination between health-care levels. PMID- 15801654 TI - [Prostate leiomiosarcoma. Report of three cases]. AB - OBJECTIVE: Leiomyosarcoma of the prostate is an uncommon neoplasm with a poor prognosis. We review the three cases of leiomyosarcoma of the prostate observed in our hospital in the last twenty years and studied their clinical follow-up, METHODS: We have found three cases and we have studied their clinical follow-up, immunohistochemical profile and ultrastructural features. RESULTS: In all cases tumor cells were positive for vimentin and also for either desmin or actin. Two cases were considered grade III sarcomas, with an aggressive course even with treatment, they died 5 and 24 months later, respectively. The third case, was considered grade II and is still alive, 60 months after diagnosis, without evidence of disease. CONCLUSIONS: Leiomyosarcoma of the prostate is an uncommon neoplasm that accounts for less than 0.1% of prostate tumors. We found no prognostic factors for predicting prolonged survival although complete resection and low mitotic activity may be predictive. PMID- 15801655 TI - [Bladder squamous metaplasia: report of one case and bibliographic review]. AB - OBJECTIVES: We report a new case of squamous metaplasia because of its interest and subtle differential diagnosis with other bladder pathologies. METHODS: We report the case of a male patient with history of previous neoplasia with an exophytic lesion of the bladder wall discovered on follow-up. Pathological diagnosis after TUR was bladder squamous metaplasia. RESULTS: Follow-up is performed by ultrasound, cystoscopy, and urine cytology in adherence to clinical guidelines due to the possibility of transformation into squamous cell carcinoma. CONCLUSIONS: Urothelium is able to develop non neoplastic transformations such as squamous metaplasia. The importance of such transformations depends on proper diagnosis and follow-up due to their ability to transform into a neoplastic process. PMID- 15801656 TI - [Update of the WHO classification of tumors of the upper urothelium]. PMID- 15801657 TI - Extramammary Paget's disease of scrotum. A case with local lymph node metastasis. AB - OBJECTIVE: We describe the clinical, histological and immunohistochemical studies in a case of extramammary Paget's disease localized in the scrotum with lymph node inguinal metastasis. METHODS/RESULTS: A 80-year old man consulted with a one year history of a pruritic erythematous skin rash of the scrotum. Physical examination showed demarcated erythematous lesion involving the scrotum and right inguinal adenopathy. Final histological examination revealed extramammary Paget's disease with inguinal metastasis. CONCLUSION: Extramammary Paget's disease of the scrotum is a rare disease. This pathological condition may spread to dermal region and the regional lymph nodes. Although genitourinary cancer may accompany scrotal extramammary Paget's disease, an extensive search for cancer may be unnecessary. PMID- 15801658 TI - [Emotional reasons for therapeutic failures]. AB - Medicine's science is still bound to a mechanistic, 17th century world-view. To give it up would force the physician to give up to manipulate the patient and to feel omnipotent toward him. Emotional reasons block him from doing so. Treatment mistakes are the consequence. The neglect of feelings evoked by the patient in the physician helps him to avoid to be exposed to own feelings, i.e. anxiety, hopelessness, anger. Again treatment mistakes follow. Sticking to theory and practice of alternative medical methods without having proved their efficiency prevents the physician to give up the cherished autistic undisciplined, magical thinking. Treatment mistakes ensue. PMID- 15801659 TI - [Experiences with "incident reporting" at the East Switzerland Childrens Hospital]. AB - Our healthcare system has become a "high-tech" industry and, as a consequence, incidents are almost "normal" events. It is one of the most important obligations in the quality management to get control of these very dangerous and therefore extremely important problems. The Children's Hospital in St. Gallen (Switzerland) introduced in 1998 a reporting system in order to monitor incidents and mistakes and to be able to introduce preventive action. For this we have been using the so called "system approach", i.e. we are not trying to find a guilty person or a single source for a mistake, but we analyze the complete working system involved in the action. According to our experience, such reporting systems have to respect a number of specific rules and prerequisites in order to be successful. The most crucial one is a "change of culture", which means, that we are not looking for a guilty person, but for the weak elements which are responsible for the mistakes in our working systems. In Switzerland a National Foundation for Patient Safety has been created for supporting all the acting professionals in the healthcare system in their challenging task. PMID- 15801660 TI - [Critical incidents]. AB - In medicine real severe mishaps are rare. On the other hand critical incidents are frequent. Anonymous critical incident reporting systems allow us to learn from these mishaps. This learning process will make our daily clinical work safer Unfortunately, before these systems can be used efficiently our professional culture has to be changed. Everyone in medicine has to admit that errors do occur to see the need for an open discussion. If we really want to learn from errors, we cannot punish the individual, who reported his or her mistake. The interest is primarily in what has happened and why it has happened and not who has committed this mistake. The cause for critical incidents in medicine is in over 80% the human factor Poor communication, work under enormous stress, conflicts and hierarchies are the main cause. This has been known for many years, therefore have already 15 years ago high-tech industries, like e.g. aviation, started to invest in special courses on team training. Medicine is a typical profession were until now only the individual performance decided about the professional career Communication, conflict management, stress management, decision making, risk management, team and team resource management were subjects that have never been taught during our preor postgraduate education. These points are the most important ones for an optimal teamwork. A multimodular course designed together with Swissair (Human Aspect Development medical, HADmedical) helps to cover, as in aviation, the soft factor and behavioural education in medicine and to prepare professionals in health care to work as a real team. PMID- 15801661 TI - [Preliminary results of an anonymous internet-based reporting system for critical incidents in ambulatory primary care]. AB - To learn from errors is not always easy, especially if they happened to others! This paper describes the organization and management of a critical incident reporting system for primary care physicians in Switzerland and reports about the difficulties and experiences during the first 18 months since the start of the program. It seems to be particularly difficult to enhance the attentiveness of physicians for apparently harmless daily critical incidents and to motivate them to report it even in an anonymous reporting system. As incentives for more intensive participation there are the hope for comments on reported cases by other participants and the expectation that reported errors will be avoided by the readers. PMID- 15801662 TI - [Failure prevention in clinical practice through identification of failure examples]. AB - "Learning from ones mistakes" is an essential part of clinical quality improvement. But focusing on a retrospective analysis of mishaps from the past is not sufficient for quality enhancements, nor is the restriction to mistakes in the solely legal sense. Analysis has to include anonymous data aggregated over dissatisfying results from multiple sources. Recognizing patterns of underlying causes for errors is the key to effective error prevention. In many cases the true cause might be predominantly organizational. PMID- 15801663 TI - [Learning from the experiences of an arbitration unit for medical liability questions]. AB - While other countries', e.g. the United States', experience with alternative dispute resolution (ADR) in medical malpractice has been disappointing, German physicians have instituted extrajudicial claims resolution as an alternative to civil litigation: Established by the Arztekammern (corporate bodies of professional self-governance) in the mid-1970s, the Schlichtungsstellen (SchlSt- claims resolution panels) offer patients and doctors, on a voluntary basis, proceedings directed toward achieving a settlement. The number of cases submitted to these panels has been constantly growing since. In 90% of cases decided upon by the panel, civil litigation was avoided. Doctors can, as exemplified by the operation of the SchlSt, tackle difficult and complex problems and solve them- without being funded or directed by outside agencies. What can be learnt from being, as a physician member, associated with a SchlSt for several years? (1) Physicians--as Eddy has pointed out--must make decisions about complex problems under difficult conditions on the basis of inadequate information. Expert testimony suffers from similar uncertainties. Decisions on individual cases cannot be improved by external and necessarily generalized ruling (disease management programs, other directives). (2) Errors and adverse events in medical care have to be accepted as principally unavoidable. Most claims heard by the panel have to do with operations and--in the non-surgical specialties--with invasive procedures. Medical tasks of high complexity, as many operative and invasive procedures indeed are, seem to have an inherent accident-proneness. Therefore decisions as to operations and invasive procedures should be weighed with utmost care. More is not always better (3) 25% of the patients filing claims with the SchlSt complain of deficient doctor-patient-communication. Failure of doctors and staff to communicate adequately with the patients and with their professional contacts plays an important role as risk factor for negligence and injury and even as cause of disputes and claims. (4) The GP, the generalist in a profession characterized by specialization, has to treat a wide spectrum of diseases, wider than any specialist. As far as can be seen, he does his job well. He needs adequate--if possible personal--contact with and support by his colleagues involved in the treatment of the same patient. PMID- 15801664 TI - [Errors in pharmacotherapy]. AB - The article discusses step by step the drug-prescription process from the assessment of a patient's medical history to the administration or intake. Practical examples illustrate common errors which could lead to complications in the inpatient as well as outpatient setting. PMID- 15801665 TI - [List of complications--reduction of errors and complications by elimination of unsafe actions and conditions]. AB - The "Komplikationenliste " (list of complications) of the association of the heads of departments of the Swiss society of internal medicine is a clinical tool for the registration and reduction of complications/errors of medical treatment. It is the aim to improve patient safety in hospital medicine. By the systematic and prospective analysis of interventions and complications in the participating medical departments it is possible to collect also rare incidents. All the complications and errors are communicated to all the departments of internal medicine in Switzerland in order to reduce them. The principle for the reduction of such events is the elimination of unsafe actions and conditions and apparently harmless errors in daily diagnostic and therapeutic interventions in order to avoid the potentially serious consequences. PMID- 15801666 TI - Beneficial effects of probiotics and prebiotics on human health. AB - The primary role of a diet is not only to provide enough nutrients to fulfill metabolic requirements of the body but also to modulate various functions of the body. Probiotics, prebiotics and synbiotics are either beneficial microorganisms or substrates that facilitate the growth of these microorganisms which can be suitably harnessed by the food manufacturers and holds considerable promise for health care industry. Regardless of sufficient health benefits of these, there is a need to carry a multidisciplinary approach on safety evaluation as the conventional toxicological approach has various limitations. This article gives an overview of probiotics and prebiotics, their health effects, mode of action, growth and survival in GIT, quality assurance criteria and safety including future prospects. PMID- 15801667 TI - [Benzo[h][1 ,6]naphthyridines from dimethyl 1,2-dihydro-2-(2-nitrophenyl) pyridine-3,5-dicarboxylate]. AB - The reaction of 2-nitrobenzaldehyde with methyl propiolate and ammonium acetate in acetic acid yields 2,6-dinor-nifedipine (1a) and as a by-product not the dimethyl 2,5-dicarboxylate isomer, but the rac. 1,2-dihydropyridine (DHP) 1b, as proven by X-ray analysis. The benzo[h][1,6]naphthyridines 4-6 are synthesized from the oxidation product 2b and the photo-product 3b. The compounds 6 represent starting materials for potential anti-malarial agents. PMID- 15801668 TI - Synthesis of some new 2-(2-fluoro-4-biphenylyl)propionic acid derivatives as potential anti-inflammatory agents. AB - The synthesis of a group of 1,3,4-oxadiazoles, 1,2,4-triazoles, 1,3,4 thiadiazoles and 1,2,4-triazine derived from 2-(2-fluoro-4-biphenylyl) propionic acid is described. The structures of new compounds are supported by IR, 1H NMR and MS data. These compounds were tested in vivo for their anti-inflammatory activity. The compounds which showed activity comparable to the standard drug flurbiprofen, were screened for their analgesic, ulcerogenic and lipid peroxidation activities. Five out of seventeen new compounds, showed very good anti-inflammatory activity in the carrageenan induced rat paw edema test with negligible ulcerogenic action. The compounds, which showed less ulcerogenic action, also showed reduced malondialdehyde production (MDA), which is one of the byproducts of lipid peroxidation. The study showed that the compounds inhibit the induction of gastric mucosal lesions and it can be suggested from our results that their protective effects may be related to an inhibition of lipid peroxidation in the gastric mucosa. PMID- 15801669 TI - Capillary electrophoretic separation of enantiomers of amino acids and amino acid derivatives using crown ether and cyclodextrin. AB - The capillary zone electrophoresis using (+)-18-crown-6-tetracarbonic acid as a chiral selector was a suitable method for the enantiomeric separation of racemates of amino acids and of some amino acid derivatives (esters, dipeptides). The influence of the chemical structure of the compounds on the separation was investigated. After optimization of the separation conditions, baseline separations were obtained for most racemates. The addition of acetonitrile and TBAB yielded an improvement of the separation. Improved selectivity was further observed by the application of a cyclodextrin, HP-beta-CD, in combination with the crown ether. PMID- 15801670 TI - The effect of the process variables on the HPLC separation of tricyclic neuroleptics on a calixarene-bonded stationary phase. AB - The chromatographic behavior of a new HPLC-stationary phase with supramolecular selectors on the basis of calixarenes is described for the separation of nine tricyclic neuroleptics. The effects of different chromatographic conditions (buffer system, pH-value, type and content of organic modifier, injection volume) on the separation of the analytes were studied. Additionally, the effect of structural differences of the neuroleptic analytes was studied. The chemical structure and pKa of the neuroleptics highly influenced their separation on the calix[8]arene phase. The separation of all analytes on the investigated calixarene-bonded stationary phase was possible with a mobile phase of acetonitrile with 30 mM ammonium acetate buffer (pH 3.5) 30:70(v/v) using 1 ml/min flow rate. PMID- 15801671 TI - Adsorptive stripping voltammetric determination of ambroxol. AB - An electrochemical procedure for the determination of ambroxol in mucolytics was described. The method was based on adsorptive accumulation of the species at the hanging mercury drop electrode (HMDE), followed by one of different modes of stripping sweep, viz. direct current tast (DCT), differential pulse (DP), square wave (SW) and first harmonic alternating current (AC1). The behaviour of adsorptive stripping response was studied under various experimental conditions, e.g. type of supporting electrolyte, pH, accumulation time, pulse amplitude, scan rate and mode of sweep. In Britton-Robinson buffer solution, an irreversible reduction process involving transfer of one electron and one proton was took place. The response was linear over the 0.2-6 microg/ml concentration range. Determination of the compound in oral dosage forms was achieved using the standard addition method. The average of determinations obtained by the squarewave adsorptive voltammetric method with its relative standard deviation was 99.8 +/- 2.40%. PMID- 15801672 TI - Initial-rate method for the determination of pantoprazole in pharmaceutical formulations using 1-fluoro 2,4-dinitrobenzene. AB - A simple and selective kinetic spectrophotometric method for the determination of pantoprazole in pharmaceutical preparations is described. The procedure is based upon a kinetic investigation of the reaction of the drug with 1-fluoro-2,4 dinitrobenzene in DMSO at room temperature. The absorbance of the coloured product was measured at 420 nm. The plot of the logarithm of the initial rate of the reaction vs. the logarithm of molar concentration of pantoprazole is linear over the range 10-20 microg x ml(-1). The procedure retains its accuracy in the presence of a large excess of its degradate, sulfenic acid, which is prepared by degradating the pure drug in borate buffer of pH 8 at room temperature for seven days. The results are validated statistically and through recovery studies. The method has been successfully applied to the determination of pantoprazole in commercial tablets. Statistical comparison of the results with the reference method shows excellent agreement and indicates no significant difference in accuracy and precision. PMID- 15801673 TI - Formation of spherical micelles composed of the novel platelet activating factor receptor antagonist, E5880. AB - E5880, a novel platelet activating factor receptor antagonist, was dispersed in water for use in an injectable formulation and the physicochemical properties of the preparation were characterized. The critical concentration for formation of micelles was 0.12 mM. Using area per molecule data, the critical packing parameter was calculated, indicating that the structure of the micelles was spherical and that each micelle containes 49 molecules. The diameter of the micelles was 8.1 nm. Attractive interactions occurred between E5880 molecules in the micelle. The hydrocarbon region in the micelle was more rigid and less hydrated than that of other surfactants, stearyltrimethylammonium chloride and cetyltrimethylammonium chloride. PMID- 15801674 TI - [Manufacturing and stability of copper-histidine solution for treatment of Menkes' Kinky Hair Syndrome]. AB - Menkes' Kinky Hair Syndrom is a rare, X-linked recessive multisystemic lethal disorder of copper metabolism. Male infants who are affected usually die at the age of 2-3 years. If the disease is diagnosed early, patients profit from subcutaneously administered copper salts. We describe the preparation and stability of a copper-histidin solution. This solution has some advantages in terms of stability over the solutions described in earlier publications. This is a great advantage for the patients and their parents, because an ambulatory care or a home care of this patients is possible now. PMID- 15801675 TI - Formulation and evaluation of an effective pH balanced topical antimicrobial product containing tea tree oil. AB - The effect of pH on the antimicrobial activity of Melaleuca alternifolia essential oil formulations was studied. Microemulsions, liposomal dispersions, multiple emulsions and a colloidal bed of sterile clay were formulated using 5% w/w of tea tree oil. A number of formulations were prepared at various pH values (5.0, 5.5, 6.0, 6.5, and 7.0). Thermal stability studies showed that the formulations were stable for more than eight months. Agar dilution tests showed MICs of 1.0% v/v S. aureus and S. epidermidis. In the broth dilution test, MBC of the oil for P. acnes was 0.5% v/v. MIC and MBC values were comparable to those of non-formulated tea tree oil, indicating that tea tree oil retained its activity in the above-mentioned formulations. The microbiological evaluation showed that the formulations containing 5% w/w tea tree oil had a maximum effect at pH 5.5. PMID- 15801676 TI - Regional permeability of coenzyme Q10 in isolated rat gastrointestinal tracts. AB - The objective of the study was to identify the region with the maximum permeability for low bioavailable coenzyme Q10 (CoQ) in the gastrointestinal tract. To evaluate the regional differences in permeability, male Sprague-Dawley rats, 250-300 g, were anesthetized and the gastrointestinal segments were isolated. Stomach, duodenum, jejunum, ileum and colon tissues were mounted on a Navicyte side-by-side diffusion apparatus. Radiolabeled CoQ (1 microM in DMEM, pH 7.4, 37 degrees C was added to the donor side and the samples withdrawn from the receiver compartment at predetermined time intervals were analyzed using a scintillation counter. Membrane integrity was monitored by 14C-mannitol permeability. The apical to basal permeability coefficients (Papp x 10(-6), cm/s) were 0.32 +/- 0.13, 3.14 +/- 0.89, 1.36 +/- 1.4, 0.83 +/- 0.40, and 1.59 +/- 0.13, for CoQ through rat stomach, duodenum, jejunum, ileum, and colon tissues respectively. The basolateral to apical permeability coefficients (Papp x 10(-6), cm/s) were 1.6 +/- 0.2, 2.2 +/- 1.2, 0.88 +/- 0.12, 1.6 +/- 0.42, and 1.9 +/- 0.41 respectively. Therefore the region of maximum CoQ permeability is duodenum followed by colon and ileum. Jejunum and stomach regions also have fairly high permeability. Therefore CoQ formulations should be made with an aim to target the duodenum to get maximum dosage effect. PMID- 15801677 TI - An attempt to clarify the influence of glycerol, propylene glycol, isopropyl myristate and a combination of propylene glycol and isopropyl myristate on human stratum corneum. AB - The present study is a comparison of the influences of glycerol, propylene glycol (PG), isopropyl myristate (IPM) and a combination of PG and IPM (1/1; w/w) on human stratum corneum (SC) by means of differential scanning calorimetry (DSC) and wide and small angle X-ray-diffraction (WAXD and SAXD). The effects of glycerol and PG on SC structure can be attributed to their functional groups. In DSC transition temperatures of lipid fractions are decreased whereas SAXD long distances of lamellar phases reveal an additional interference due to an integration into hydrophilic regions of hexagonally packed lipids (PG) or orthorhombically packed lipids (glycerol). The increased repeat distance is attributed to the polar character of both molecules. However, with IPM the long distance remains unaffected. IPM is integrated into the lipophilic regions of SC lipid matrix as concluded from an increase of WAXD reflections of orthorhombical lipids and a decrease of WAXD reflections of hexagonal lipids. The combination of PG/IPM affects SC microstructure in a specific manner. DSC shows a decrease in transition temperatures of the lipid fractions, although not as much as expected from the single substances. Additionally, the combination of IPM/PG affects the short distances of orthorhombically and hexagonally packed lipids in WAXD measurements similar as PG alone, whereas the long distance seems to remain unaffected as in the case of IPM pretreatment. Adjuvants with penetration enhancing potential reveal different effects on SC lipid microstructure, which have to be kept in mind in terms of formulating systems for transdermal administration. PMID- 15801678 TI - Stereoselectivity and interaction between the glucuronidation of S-(-)- and R-(+) propranolol in rat hepatic microsomes pretreated with different inducers. AB - Phase II glucuronidation metabolism of side-chain propranolol was studied using microsomes from rats treated with the inducers beta-naphthoflavone (BNF) or dexamethasone (Dex). The glucuronide concentrations of propranolol enantiomers were assayed by RP-HPLC. The kinetic constants of glucuronidation, Km, Vmax and Clint were determined. There are significant differences between the R- and S enantiomeric glucuronide in Km, Vmax and Clint P < 0.05, P < 0.01 and P < 0.05 in control microsome. There are significant differences in Km and Clint (P < 0.01 or P < 0.001) but no significant differences in Vmax (P > 0.05) between R and S enantiomeric glucuronide in the microsomes induced with Dex and BNF. The formation of S-(-)-propranolol glucuronide was inhibited by R-(+)-propranolol from the rat microsomes pretreated with BNF and Dex. The glucuronidation metabolism of propranolol enantiomers exhibited the stereoselectivity in rat hepatic microsomes induced with BNF or Dex. Multiple UGT1A and 2B may be involved in stereoselective O-glucuronidation of propranolol enantiomers in rat liver microsomes. The glucuronides produced were in favor of the R-enantiomer. There is an interaction between the glucuronidation of R- and S-enantiomer. PMID- 15801681 TI - A new flavone glycoside and other constituents from Carduus crispus. AB - A new flavone glycoside, chrysoeriol 7-O-(2"-O-6'''-O-acetyl-beta-D glucopyranosyl-beta-D-glucopyranoside (1), along with fourteen known compounds 2 15 were isolated from the whole plant of Carduus crispus Linn. Their structures were established on the basis of spectroscopic methods and chemical evidences. The antitumour activity of compound 1, 4 and 5 was tested. Compound 4 exhibited significant antitumour activity against HO-8901 (human ovarian neoplasm) cells. PMID- 15801680 TI - Influence of Casearia esculenta root extract on glycoprotein components in streptozotocin diabetic rats. AB - The present study was aimed to evaluate the role of the indigenous antidiabetic medicinal plant Casearia esculenta on glycoprotein components in streptozotocin induced diabetic rats in plasma, liver, kidney and cardiac tissues. Streptozotocin injection (50 mg/kg body weight) caused massive elevation of glycoprotein components such as hexose, hexosamine, sialic acid and fucose in plasma and tissues of diabetic control and experimental animals. Oral administration of C. esculenta root extract (200 and 300 mg/kg body weight) for 45 days significantly reverted the hexose, hexosamine, sialic acid and fucose levels to near normal values. These results suggest a normalizing effect of C. esculenta on glycoprotein components in STZ diabetic rats. PMID- 15801682 TI - Coulometric titration of ketotifen in tablets. AB - A method for the determination of ketotifen involving its reaction with iodine in an alkaline medium is presented. In coulometric titration using biamperometric end-point detection 0.25-2 micromol (77-618 microg) of ketotifen was successfully determined. The elaborated method was applied to the determination of ketotifen in drugs. PMID- 15801679 TI - 5-(3-Phenylpropoxy)psoralen and 5-(4-phenylbutoxy)psoralen: mechanistic studies on phototoxicity. AB - 5-(3-Phenylpropoxy)psoralen and 5-(4-phenylbutoxy)psoralen are blockers of the lymphocyte potassium channel Kv1.3 with EC50 values < 10 nM and may be used as new potential immunosuppressive drugs. As reported for many furocoumarins, these compounds show phototoxic properties and react under UV radiation with different targets in the cell, e.g. DNA or proteins and lipids in membranes. The photoreactions with these targets were investigated and compared to the well known derivatives psoralen, 5-methoxypsoralen, 8-methoxypsoralen and 4,5',8 trimethylpsoralen. Moreover, absorption properties and the capability to photoproduce singlet oxygen (1O2) and reactive oxygen species (ROS) were studied. It was found that 5-(3-phenylpropoxy)psoralen and 5-(4-phenylbutoxy)psoralen are similar or less phototoxic in vitro as 5-methoxypsoralen which is the furocoumarin with the weakest phototoxicity in vivo compared to psoralen, 8 methoxypsoralen and 4,5',8-trimethylpsoralen. PMID- 15801683 TI - Surface modification of liposomes for cardiomyocytes targeting in vitro. AB - The effect of novel 3-{4-[2-hydroxyl-(1-methyl ethylamine) propyl oxygen]phenyl}propionic acid cetylester (PAC) as a surface modification ligand on the delivery of liposomes into cultured cardiomyocytes was investigated. Small unilamellar liposomes with and without PAC (PAC-liposome and Plain-liposome) were labeled with a fluorescence marker. The cultured neonatal cardiomyocytes were incubated with liposomes under normoxia or hypoxia conditions, and then the cell associated fluorescence was measured. A high affinity of the PAC-liposomes to cardiomyocytes was observed. The amount of cell uptake of PAC-liposomes under normoxia conditions was 4-fold higher than that of plain-liposome, and the increase was 8.5-fold when hypoxia occured. The results suggested that PAC is a potential surface modification ligand for liposome targeting the ischemic myocardium. PMID- 15801684 TI - The endothelial cells in islets of langerhans. AB - The blood vessels of the pancreatic islets are of crucial importance for oxygen and metabolite supply, and dispersal of secreted hormones. In addition to this, endothelial cells have an important role in the revascularization process after islet transplantation. Studies have reported signs of poor engraftment of transplanted islets, presumably due to impaired revascularization. The aims of this study were to investigate islet endothelial cells and the revascularization process of transplanted islets. The lectin Bandeiraea simplicifolia was found to consistently stain endothelium of both endogenous and transplanted pancreatic islets. By using this marker, we investigated the vascular density of both endogenous and transplanted islets of C57BL/6 mice. One month post transplantation, a time point when the implants are assumed to be completely revascularized, the graft vascular density was decreased at all investigated implantation sites when compared to endogenous islets. Furthermore, most of the blood vessels were located in the graft connective tissue stroma. Similar results were obtained six months post-transplantation and in cured diabetic animals after one month. In order to evaluate the function of intraportally transplanted islets, we developed a method to retrieve such islets. Enzymatic and mechanic treatment of the liver enabled us to re-isolate the transplanted islets for further in vitro studies. These islets had decreased insulin release, insulin content and glucose oxidation rate when compared to non-transplanted control islets. To understand the role of islet endothelium in the revascularization of transplanted islets we performed angiogenesis microarray studies on islet endothelial cells, from non-cultured, cultured and transplanted islets. We found that the islet endothelium expressed mRNA for both inhibitors and inducers of angiogenesis, and that this expression differed with time. In conclusion, these results provide a useful platform for further studies on the islet endothelium. PMID- 15801685 TI - The role of the innate immunity in islet transplantation. AB - Clinical islet transplantation is an emerging procedure to cure type 1 diabetes. The graft is implanted by infusion into the liver through the portal vein. A major obstacle that still needs to be overcome is the requirement for islets from multiple donors to achieve insulin independence. An innate inflammatory reaction, the IBMIR, is elicited when islets are exposed to blood. The IBMIR has been described as a clotting reaction culminating in disruption of islet morphology and is a plausible cause for loss of tissue during the early post-transplant period. In this thesis, the underlying mechanisms of the IBMIR were characterized. The IBMIR was for the first time demonstrated in patients undergoing an islet transplant, and a number of clinically applicable strategies to limit this reaction were identified. The thrombin inhibitor melagatran completely blocked the IBMIR in an in vitro tubing blood loop system, indicating that thrombin is the driving force in the reaction. Interestingly, islets were shown to produce and secrete tissue factor (TF), the physiological trigger of coagulation. Inactivated FVIIa, a specific inhibitor of TF, successfully blocked initiation of the IBMIR. An alternative approach to limit the IBMIR was to pre treat islets in culture prior to transplantation. Nicotinamide added to the culture medium effectively decreased the level of TF in human islets. Infiltration of immune cells, also a part of the IBMIR, was characterized in detail. The predominant cell types infiltrating the islets were neutrophilic granulocytes and, to a lesser degree, monocytes. Both cell types may exert direct cytotoxic effects, and the antigen-presenting monocytes may also be important for directing the specific immune system to the site of inflammation. These findings have provided new insight into the nature of the IBMIR and offer several new strategies to improve the outcome of clinical islet transplantation. PMID- 15801686 TI - Changes in graft blood flow early after syngeneic rat pancreas-duodenum transplantation. AB - Organ transplantation is associated with changes in graft blood flow, both acutely caused by reperfusion associated phenomena, and chronically due to e.g. denervation. The aim of the study was to investigate regional blood flow early after implantation of a syngeneic pancreas-duodenum transplant in rats, i.e. during reperfusion. Warm ischemia time was 1-2 min and cold ischemia 90 min. Blood flow values were measured with coloured microspheres both 10 and 30 min after implantation in transplanted rats, and at one time point in control rats. A marked decrease in the blood perfusion of the transplanted duodenum compared to the endogenous intestine was seen at both 10 and 30 min. Total graft pancreatic blood flow was increased both 10 and 30 min after implantation, whilst islet blood flow remained unchanged compared to the endogenous gland. We conclude that the blood perfusion of the graft is markedly changed in the immediate post transplantation period, presumably due to reperfusion. However, islet blood perfusion remains constant, suggesting that islet vasculature is less sensitive to changes induced by the implantation. PMID- 15801687 TI - Lasting improvement of hyperglycaemia and bodyweight: low-carbohydrate diet in type 2 diabetes--a brief report. AB - In two groups of obese patients with type 2 diabetes the effects of 2 different diet compositions were tested with regard to glycaemic control and bodyweight. A group of 16 obese patients with type 2 diabetes was advised on a low-carbohydrate diet, 1800 kcal for men and 1600 kcal for women, distributed as 20 % carbohydrates, 30 % protein and 50 % fat. Fifteen obese diabetes patients on a high-carbohydrate diet were control group. Their diet, 1600-1800 kcal for men and 1400-1600 kcal for women, consisted of approximately 60 % carbohydrates, 15 % protein and 25 % fat. Positive effects on the glucose levels were seen very soon. After 6 months a marked reduction in bodyweight of patients in the low carbohydrate diet group was observed, and this remained one year later. After 6 months the mean changes in the low-carbohydrate group and the control group respectively were (+/-SD): fasting blood glucose (f-BG): -3.4+/-2.9 and -0.6+/ 2.9 mmol/l; HBA1c: -1.4+/-1.1 % and -0.6+/-1.4 %; Body Weight: -11.4+/-4 kg and 1.8+/-3.8 kg; BMI: -4.1+/-1.3 kg/m_ and -0.7+/-1.3 kg/m_. Large changes in blood glucose levels were seen immediately. CONCLUSION: A low-carbohydrate diet is an effective tool in the treatment of obese patients with type 2 diabetes. PMID- 15801688 TI - Skeletal muscle metastasis secondary to cancer--a report of seven cases. AB - Cancer metastasis to the skeletal muscle is very rare. In this report, we describe 7 cases (6 males, 1 female, age 55-76, mean 66 y.o.) with unusual presentation of cancer metastasis to the skeletal muscle, illustrating the principal clinical, radiographic and histologic features. The primary tumors were lung cancer in 4 cases (3 adenocarcinomas, 1 squamous cell carcinoma) and renal cell carcinoma, esophageal carcinoma, hepatocellular carcinoma in each one. The metastatic sites were the adductors, iliopsoas, rectus femoris, biceps bracii, triceps bracii, deltoid, and paravertebral muscle in each one. The unusual sites of metastases were visualized by CT and/or MRI and confirmed by needle or trocar biopsy. Five patients died in 2 to 24 months with the average of 8 months after initial examination. PMID- 15801689 TI - Infantile fibrosarcoma of thigh--a case report. AB - Infantile fibrosarcoma is a rare soft tissue malignant tumor, when it occurs, it is usually seen in the first year of life. The clinical course of infantile fibrosarcoma is more favorable and metastasis is rare compared with that in adulthood. While adult fibrosarcoma are common in the thigh, infantile fibrosarcoma affect chiefly the distal portions of the extremities. Standard treatment is primarily wide surgical excision. In this case report, we present our experience of an infantile fibrosarcoma of thigh with good clinical course 36 months after tumor resection and the usefulness of detecting the ETV6-NTRK3 gene fusion in differential diagnosis. PMID- 15801690 TI - [Roll back?]. PMID- 15801691 TI - [De-bureaucratization of nursing documentation: support instead of replacement of the practice]. PMID- 15801692 TI - [Where is the voice of nursing?]. PMID- 15801693 TI - [BVMed Safety Forum: 1 year TRBA 250--what was achieved?]. PMID- 15801694 TI - [Occupational HIV exposure by injuries: what to do after contact with infected materials?]. PMID- 15801695 TI - [Relational nursing in inpatient psychotherapy: facilitating healing experience]. PMID- 15801696 TI - [Nursing diagnosis in the general practice test: opportunity for new thoughts- and management]. PMID- 15801697 TI - [Electronic health record--(not) a topic for nursing? Nurses need access to all data (interview by Katrin Balzer)]. PMID- 15801698 TI - [Functional health and nursing performance: WHO classification challenge]. PMID- 15801699 TI - [Evidence-based nursing in ulcus cruris venosum: proper compression therapy is the key]. PMID- 15801700 TI - [Diagnosis related groups: increased profit by nursing related diagnosis of comorbidity]. PMID- 15801701 TI - [Clinical treatment pathways--1: High quality at low cost--an incompatible goal conflict?]. PMID- 15801702 TI - [Optimizing nursing documentation: basis for good nursing care]. PMID- 15801703 TI - [Hospital internal nursing process assessment: reinforcing the know-how of nursing]. PMID- 15801704 TI - [Living with multiple sclerosis: "never give up" (interview by Olivia Scheinert, Kristin Banka, Astrid Schmid, Manfred Beathalter)]. PMID- 15801705 TI - [To represent needs of nursing care using nursing diagnoses: potentials and restrictions of the NANDA classification and ICNP]. AB - Nursing diagnoses represent individual reactions to existing or potential changes in one's state of health. They are result of a diagnostic process, which is part of the dynamic nursing care process in its whole. Thus, as a basis of nursing interventions diagnoses have to be proved continuously. The classification of the North American Nursing Diagnosis Association (NANDA) as well as the International Classification for Nursing Practice (ICNP) can be account to the international well-known classifications of nursing diagnoses. Comparing their structures, some fundamental differences between both classifications become obvious. While the NANDA classification represents a systematic structured body of nursing knowledge with regard to human health reactions patterns, the ICNP reflects a more comprehensive part of the nursing reality, since it also contains nursing interventions and outcomes. Until the latest changes by establishing the taxonomy II, NANDA diagnoses have primarily focused deficits. But in contrast to the diagnoses of the ICNP they also comprise etiological factors. To prove the applicability of both classifications to nursing practice, they have been applied to a case study of a female resident living in a nursing home. The results of analysis show that because of their different structures the NANDA classification and ICNP have their own possibilities and limitations in covering the resident's individual needs of nursing care. These characteristic potentials and restrictions have to be taken into account when one of the classification systems is going to be implemented into nursing practice. PMID- 15801706 TI - When the computers crash. PMID- 15801707 TI - A physician's guide to CLIA rules. PMID- 15801708 TI - My patient did it her way. PMID- 15801709 TI - Extend your practice-not your liability. PMID- 15801710 TI - Is peer review worth saving? PMID- 15801711 TI - Dr. Ulrich's battle. PMID- 15801713 TI - Forced charity isn't charity at all. PMID- 15801712 TI - When a patient won't pay. PMID- 15801714 TI - PEGylated immunoliposomes directed against brain astrocytes. AB - Polyethylene glycol (PEG)ylated (stealth) immunoliposomes directed against human gliofibrillary acidic protein (GFAP) were prepared by coupling the thiolated monoclonal anti-GFAP antibodies with a maleimide derivative of phosphatidyl ethanolamine of the liposomal membrane. Experiments with cell cultures demonstrated specific and competitive binding of these immunoliposomes to embryonic rat brain astrocytes. Administered intravenously into rats, the immunoliposomes displayed typical kinetics with elimination half-lives of 8-15 hr. Being incapable of penetrating the unimpaired blood-brain barrier (BBB), these immunoliposomes, nevertheless, may be useful in delivering drugs to glial brain tumors (which continue to express GFAP) or to other pathological loci in the brain with a partially disintegrated BBB. PMID- 15801715 TI - In vitro transcutaneous delivery of ketoprofen and essential polyunsaturated fatty acids from a fish oil vehicle incorporating 1,8-cineole. AB - Transcutaneous administration of nonsteroidal anti-inflammatory drugs and essential fatty acids from fish oil, principally eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), may simultaneously lead to increased cyclooxygenase inhibition and the production of less potent inflammatory mediators within joints. The objective of our study was to determine the permeation of ketoprofen, EPA, and DHA (from fish oil) across pig ear skin in vitro in the presence of the enhancer 1,8-cineole. Formulations containing 2.5% ketoprofen in fish oil with varying concentrations of 1,8-cineole were prepared and applied to full-thickness pig ear skin mounted in all glass Franz-type diffusion cells. Simultaneous permeation of ketoprofen and EPA and DHA from these formulations was determined by reverse phase HPLC over a 48-hr period (n = 6). We found that fish oil alone enhanced the permeation of ketoprofen across pig ear by a factor of 1.72 relative to a water vehicle. There was a dose-dependent increase in the rate of permeation of ketoprofen relative to the concentration of 1,8-cineole. The highest Q24 and Q48 was obtained with a 20% 1,8-cineole formulation with values of 355.78 +/- 50.73 microg cm(-2) and 963.29 +/- 136.69 microg cm(-2), respectively. Surprisingly, no clear effect upon the permeation of EPA and DHA by 1,8-cineole was observed, with the highest Q24 and Q48 values seen in a formulation containing no 1,8-cineole. This may have been due to differential solvation effects prior to or during the permeation process or modulation of the skin during the permeation process. PMID- 15801716 TI - Modified doxorubicin for improved encapsulation in PVA polymeric micelles. AB - Polyvinylalcohol, partially substituted with lipophilic acyl chains, generates polymeric micelles in aqueous phase, containing a hydrophobic core able to encapsulate lipophilic drugs. Two types of polymers were obtained by conjugation of polyvinylalcohol with oleoyl or linoleoyl chains as pendant groups. The polymers, at a substitution degree of approximately 1%, are soluble in water and form polymeric micelles whose size increases with polymer concentration. Doxorubicin was hydrophobized, by linking an oleoyl chain via amide bond, to make the drug more similar to the substituted polymers and promote its encapsulation into the inner core of the micelles. The properties of the drug-polymer systems were evaluated in solution by dynamic light scattering technique and correlated to the physicochemical characteristics of the drug and the substituted polymers. Solubilization tests revealed that the similarity of the chain, in both the polymer and the drug, promotes better drug encapsulation in the oleoyl than linoleoyl derivative. The drug-polymer systems are stable in phosphate buffer saline (pH 7.4) at 37 degrees C, and the release of the drug is activated by the presence of the proteolytic enzyme pronase-E. The enzyme activated drug release and the size of the polymeric micelles, compatible with the pore dimensions of the tumor vessels, make these systems interesting for targeting lipophilic drugs to solid tumors, where the proteolytic enzyme concentration strongly raises with respect to the other body compartments. PMID- 15801717 TI - Micelles based on polyvinyl alcohol substituted with oleic acid for targeting of lipophilic drugs. AB - Polymeric micelles based on polyvinyl alcohol substituted with oleic acid were used as vehicles for progesterone and folic acid. The ability of this amphiphilic polymer to entrap lipophilic drugs and to generate stable micelles in aqueous neutral medium makes it a good candidate for drug delivery. The release of the loaded drugs in acidic environments represents another important property of these systems. Size of micelles, their stability, and their drug-loading capacity were evaluated, as well as the in vitro controlled-release profiles at pH 7.4 and 5.5. PMID- 15801718 TI - Caco-2 cell culture as a model for famotidine absorption. AB - The aim of the study was to determine penetration properties of Famotidine fro the formulations through colon adenocarcinoma (Caco)-2 cell monolayers and to compare in vitro with in vivo test results. It also aimed to determine the effect of particle size on the penetration properties of Famotidine when microsphere formulations were used. Famotidine was chosen as a model drug and Caco-2 cell culture model was used. Biodegradable Famotidine microspheres of poly(lactide-co glycolide)(PLGA) polymer (50:50) were prepared by using multiple emulsion technique. Microspheres were coded according to their particle size and polymer[LHIV:60 microm Famotidine-PLGA(high viscosity), SHIV:6 microm Famotidine PLGA(high viscosity), LLIV:60 microm Famotidine-PLGA (low viscosity), SLIV:6 microm Famotidine-PLGA (low viscosity)]. Famotidine solution(5 mg/ml) and microsphere formulations were administered orally to mice and blood drug levels were determined and compared with the Caco-2 cell experiments. Permeability values of Famotidine through Caco-2 cells from various formulations were determined (log k(solution) = 7,274 +/- 0,010,log kSHIV = -3,884 +/- 0,033,log kLHIV = -2,300 +/- 0,009,log kSLIV = -4,076 +/- 0,208,log kLLIV = 3,525 +/- 0,045). Our results showed that H2 receptor antagonists alter the barrier properties of the Caco-2 cell monolayer by causing an increment in the tightness of the tight junctions. Therefore, amount of the H2 receptor antagonist-like drug at the site of action was found to be important as well as polymer type and particle size of microspheres for drug permeation. Permeation of the drug was lower when higher amounts of Famotidine were present at the diffusion site. A controlled release dosage form of H2 receptor antagonist-like drugs may be beneficial for long-term treatments. PMID- 15801719 TI - Solubilization and pharmacokinetic behaviors of sodium cholate/lecithin-mixed micelles containing cyclosporine A. AB - The purpose of this study was to investigate the solubilization capacity of sodium cholate/lecithin-mixed micelles and to evaluate the potential of mixed micelles as a carrier of cyclosporine A for intravenous infusion. The mixed micelles were prepared by coprecipitation technique. The formulation components and preparation procedures, which may affect the solubilization of cyclosporine A, were studied. The dilution stability of cyclosporine A-containing mixed micelles was investigated. Pharmacokinetic behaviors of mixed micelles in rabbits after intravenous infusion were compared with Sandimmun. Results showed the strategies to increase the solubility of cyclosporine A include lowering the molar ratio of sodium cholate to lecithin, increasing the concentration of lecithin, and reducing the ionic strength of the dispersion medium and temperature. The largest solubility was found to be 5.42 +/- 0.16 mg/ml. The leakage of mixed micelles in 5% glucose (5.84%) was much less than that in saline solution (36.7%). The relative bioavailability of mixed micelles versus Sandimmun was 112 +/- 20%, and statistical analysis demonstrated both preparations were bioequivalent. Sodium cholate/lecithin-mixed micelles are promising carriers in the intravenous delivery of cyclosporine A, considering their capability of large scale production and low-toxic property. PMID- 15801720 TI - Chitosan-based particles as controlled drug delivery systems. AB - Chitosan, a natural-based polymer obtained by alkaline deacetylation of chitin, is nontoxic, biocompatible, and biodegradable. These properties make chitosan a good candidate for conventional and novel drug delivery systems. This article reviews the approaches aimed to associate bioactive molecules to chitosan in the form of colloidal structures and analyzes the evidence of their efficacy in improving the transport of the associated molecule through mucosae and epithelia. Chitosan forms colloidal particles and entraps bioactive molecules through a number of mechanisms, including chemical crosslinking, ionic crosslinking, and ionic complexation. A possible alternative of chitosan by the chemical modification also has been useful for the association of bioactive molecules to polymer and controlling the drug release profile. Because of the high affinity of chitosan for cell membranes, it has been used as a coating agent for liposome formulations. This review also examines the advances in the application of chitosan and its derivatives to nonviral gene delivery and gives an overview of transfection studies that use chitosan as a transfection agent. From the studies reviewed, we concluded that chitosan and its derivatives are promising materials for controlled drug and nonviral gene delivery. PMID- 15801721 TI - Validation of accuracy of enzyme-linked immunosorbent assay in hybridoma screening and proposal of an improved screening method. AB - The 96-well plate format of enzyme-linked immunosorbent assay (ELISA) is the de facto standard in screening hybridomas for active antibody. Despite its widespread use, there have been few or no systematic attempts to validate its accuracy and answer the fundamental question, is it finding all the positives? We report here on a comparison between ELISA and a semiautomated flow-based kinetic exclusion assay (KinExA), both used in screening the same hybridoma cell line. Our finding is that ELISA is both overreporting (false positives) and underreporting (false negatives) compared to the KinExA system. The large number of hybridoma cells (e.g., cultured in six 96-well plates) that must be checked is daunting in considering any method other than ELISA for routine screening. To overcome this, we devised a sampling strategy in which wells are combined in a specified pattern, allowing a significant reduction in the total number of measurements required. PMID- 15801722 TI - Kinetics of anion transfer across the liquid | liquid interface of a thin organic film modified electrode, studied by means of square-wave voltammetry. AB - The electrochemical oxidation of lutetium bis(tetra-tert-butylphthalocyaninato) (LBPC) and decamethylferrocene (DMFC), as well as the reduction of LBPC, lutetium bis(phthalocyaninato) (LPC), and lutetium (tetra-tert-butylphthalocyaninato hexadecachlorphthalocyaninato) (LBPCl), has been studied in a thin nitrobenzene (NB) film deposited on the surface of a graphite electrode (GE) by means of square-wave voltammetry (SWV). The organic film-modified electrode was immersed in an aqueous (W) electrolyte solution and used in a conventional three-electrode configuration. When the aqueous phase contains ClO4-, NO3-, or Cl- (ClO4-, or NO3 only, in the case of DMFC), both LBPC and DMFC are oxidized to stable monovalent cations in the organic phase. The electron transfer at the GE | NB interface is accompanied by a simultaneous anion transfer across the W | NB interface to preserve the electroneutrality of the organic phase. LBPC, LPC, and LBPCl are reduced to stable monovalent anions accompanied by expulsion of the anion of the electrolyte from the organic into the aqueous phase. In all cases, the overall electrochemical process comprises simultaneous electron and ion transfer across two separate interfaces. Under conditions of SWV, the overall electrochemical process is quasireversible, exhibiting a well-formed "quasireversible maximum" that is an intrinsic property of electrode reactions occurring in a limiting diffusion space. For all the redox compounds that have been studied, the kinetics of the overall electrochemical process is controlled by the rate of the ion transfer across the liquid | liquid interface. Based on the quasireversible maximum, a novel and simple methodology for measuring the rate of ion transfer across the liquid | liquid interface is proposed. A theoretical background explaining the role of the ion-transfer kinetics on the overall electrochemical process at the thin organic film modified electrode under conditions of SWV is presented. Comparing the positions of the theoretical and experimental quasireversible maximums, the kinetics of ClO4-, NO3-, and Cl- across the W | NB interface was estimated. The kinetics of the overall process at the thin organic film modified electrode, represented by the second-order standard rate constant, is 91 +/- 8, 90 +/- 4, and 133 +/- 10 cm(4) s(-1) mol(-1), for the transfer of ClO4-, NO3-, and Cl- respectively. PMID- 15801723 TI - Enzyme immobilization in porous silicon: quantitative analysis of the kinetic parameters for glutathione-S-transferases. AB - Porous silicon matrixes are attractive materials for the construction of biosensors and may also have utility for the production of immobilized enzyme bioreactors. In an effort to gain a quantitative understanding of the effects of immobilization on enzyme activity, we compared the activity of glutathione-S transferase immobilized in electrochemically etched porous silicon films (approximately 6.5 microm thick) with the enzyme in solution. Kinetic measurements were made by varying the glutathione concentration while maintaining a fixed saturating concentration of 1-chloro-2,4-dinitrobenzene. The reaction kinetics follow steady-state equilibrium behavior. The specific activity of the free enzyme in solution is approximately 4x higher than the immobilized enzyme, for which we measured an apparent K'(m)(GSH) value of 1.0 +/- 0.3. The maximum velocity, V'(max), is linearly proportional to immobilized enzyme concentration, but the magnitude is approximately 20 times lower than that in solution. Results suggest approximately 25% of the enzyme is bound with the catalytic site in an inactive conformation or in a hindered orientation. Finally, the effects of hydration and exposure to denaturants on the immobilized enzyme activity are presented. PMID- 15801724 TI - Analysis of CheA histidine phosphorylation and its influence on protein stability by high-resolution element and electrospray mass spectrometry. AB - A combination of electrospray mass spectrometry (ESI-MS) and element mass spectrometry (ICPMS) with phosphorus detection was used to characterize histidine phosphorylation (His-48) of the chemotaxis protein CheA quantitatively. The phosphorylation at His-48 was found to be responsible for a stabilization of the protein. For this investigation, the acceptor domain and the kinase domain of the bacterial chemotaxis protein CheA were recombinantly expressed as single proteins. Using in vitro kinase assay conditions, the acceptor domain CheA-H was phosphorylated by the kinase domain CheA-C. The degree of histidine phosphorylation was determined by nanoelectrospray mass spectrometry of intact CheA-H, and was found to be limited to a maximum value of approximately 50%. The site specificity of CheA-H phosphorylation was controlled by nanoESI-MS/MS of the [M + 16H](16+) ion of intact (pHis)-CheA-H and allowed localization of the pHis residue to the region between residues 32 and 86, containing candidates His-48 and His-67, for which His-48 phosphorylation has been described. Analysis of the tryptic digest of in vitro histidine-phosphorylated CheA-H by capillary chromatography coupled to ESI-MS and to ICPMS with phosphorus detection revealed a truncated (pHis)-CheA-H protein as the only phosphorus-containing analyte. Since the truncated (pHis)-CheA-H in the digest was found to exhibit a higher degree of phosphorylation than could be generated by in vitro phosphorylation without trypsin treatment, it is concluded that histidine phosphorylation at His 48 strongly interferes with structural properties of the CheA-H domain in particular with respect to proteolytic degradation by trypsin. PMID- 15801725 TI - DNA aptamer-based bioanalysis of IgE by fluorescence anisotropy. AB - A rapid, homogeneous aptamer-based bioanalysis is reported for the sensitive detection of immunoglobulin E (IgE) using fluorescence polarization (FP). 5'-End labeled D17.4 DNA aptamer was used for IgE detection based on the anisotropy differences of the labeled ligand. Two different fluorophores, fluorescein and Texas Red, were used to analyze IgE in the low-nanomolar range with high specificity. Measurable anisotropy changes were observed with a short equilibration time. Analysis of the binding data reveals a possible cooperative binding process in solution. The nature of the fluorophore clearly influences the sensitivity of the analysis more than the tether length used for the dye conjugation. The local fluorophore motion is seen to influence the sensitivity of the FP probe significantly. Texas Red is seen to be relatively more sensitive for this approach and has apparently favorable dye-DNA interactions, and a limit of detection of 350 pM was obtained. Significant temperature dependence of the FP responses has been observed in this work. Ionic composition of the binding buffer also influences the assay sensitivity. The results confirm the promise and potential of similar homogeneous assays for aptamer-based bioanalysis. PMID- 15801726 TI - Effect of model ligands on iron redox speciation in natural waters using flow injection with luminol chemiluminescence detection. AB - The effects of dissolved organic compounds on the determination of nanomolar concentrations of Fe(II) have been compared using two luminol-based flow injection chemiluminescence (FI-CL) methods. One used the direct injection of sample into the luminol reagent stream, and the other incorporated on-line solid phase extraction of the analyte on an 8-hydroxyquinoline microcolumn. The CL signals from analyses of dissolved iron species (Fe(II) and Fe(III)) with model ligands and organic compounds were examined in high-purity water and seawater. The organic compounds included natural reducing agents (e.g., ascorbic acid), nitrogen sigma-donor/pi-acceptor compounds (e.g., 1,4-dipyridine, protoporphyrin IX), aromatic compounds (e.g., 1,4-dihydroxybenzene), synthetic iron chelators (e.g., EDTA), and natural iron binding compounds (e.g., desferrioxamine B, ferrichrome A). Fe(II) determinations for both luminol FI-CL methods were affected by submicromolar concentrations of redox-active compounds, strong iron binding ligands (i.e., log K(FeL) > 6), and compounds with electron-donating functional groups in both high-purity water and seawater. This was due to reactions between organic molecules and iron species before and during analysis, rather than chemiluminescence caused by the individual organic compounds. In addition, the effects of strong ligands and size speciation on Fe(II) recoveries from seawater following acidification (pH 2) and reduction (100 microM sodium sulfite) were investigated. PMID- 15801727 TI - Quantitative analysis of 39 polybrominated diphenyl ethers by isotope dilution GC/low-resolution MS. AB - A GC/low-resolution MS method for the quantitative isotope dilution analysis of 39 mono- to heptabrominated diphenyl ethers was developed. The effects of two different ionization sources, electron impact (EI) and electron capture negative ionization (ECNI), and the effects of their parameters on production of high-mass fragment ions [M - xH - yBr](-) specific to PBDEs were investigated. Electron energy, emission current, source temperature, ECNI system pressure, and choice of ECNI reagent gases were optimized. Previously unidentified enhancement of PBDE high-mass fragment ion [M - xH - yBr](-) abundance was achieved. Electron energy had the largest impact on PBDE high-mass fragment ion abundance for both the ECNI and EI sources. By monitoring high-mass fragment ions of PBDEs under optimized ECNI source conditions, quantitative isotope dilution analysis of 39 PBDEs was conducted using nine (13)C(12) labeled PBDEs on a low-resolution MS with low picogram to femtogram instrument detection limits. PMID- 15801728 TI - Headspace water-based liquid-phase microextraction. AB - A new organic-free mode of headspace liquid-phase microextraction termed headspace water-based liquid-phase microextraction has been developed. Five phenols were used as model compounds in the development and evaluation of the procedure. Parameters affecting the extraction efficiency were investigated. Under optimal extraction conditions, very high enrichment factors were achieved. Since only water (with sodium hydroxide to adjust the pH) and a conventional microsyringe were used, this new technique is affordable, efficient, and convenient for extraction of volatile and semivolatile ionizable compounds. In addition, in combination with capillary electrophoresis, the entire analytical process is totally organic solvent-free and is thus environmentally friendly. PMID- 15801729 TI - Chiral stationary phase based on a biostable L-RNA aptamer. AB - An immobilized anti-L-arginine d-RNA aptamer, used as a target-specific chiral stationary phase (CSP), was found to be very quickly degraded by RNases under usual chromatographic utilization and storage. To overcome this severe limitation for a practical use, a CSP based on the L-RNA aptamer, that is, the mirror image of the D-RNA aptamer, was created. It was shown that this mirror-image approach was a very simple and powerful strategy to develop a highly stable stationary phase due to the intrinsic insensitivity of l-RNA to the RNase degradation. In addition, such an approach allowed one to reverse the enantiomer elution order relative to that obtained with the corresponding d-RNA CSP. PMID- 15801730 TI - Photoresponsive liquid membrane transport of alkali metal ions using crowned spirobenzopyrans. AB - Several azacrown ether derivatives, which are monoaza-12-crown-4, -15-crown-5, and -18-crown-6 and diaza-12-crown-4 and -18-crown-6, bearing one or two spirobenzopyran(s), which we call crowned spirobenzopyran or crowned bis(spirobenzopyran), were synthesized and were used as carriers for liquid membrane transport of alkali metal ions. The passive alkali metal transports through liquid membranes containing crowned spirobenzopyrans were carried out under dark, and UV- and visible-light irradiation conditions. The metal ion transport was accelerated and retarded by UV- and visible-light irradiation, respectively. On the other hand, the photoresponse of the metal ion selectivity in membrane transport by crowned spirobenzopyrans was different, depending on the kind of crown ether units. Especially, diaza-12-crown-4-bis(spirobenzopyran) exhibited an excellently selective and effective transporting ability for Li(+). The uphill transports of Li(+) through a liquid membrane containing monoaza-12 crown-4-spirobenzopyran or diaza-12-crown-4-bis(spirobenzopyran) were realized under the conditions where the same aqueous solution was used as the source and receiving phases with UV and visible lights being irradiated onto the boundary phases between the source and membrane phases and between the receiving and membrane phases, respectively. The uphill transport of Li(+) from the source to receiving phases through a liquid membrane containing a crowned spirobenzopyran was also attained by the proton-concentration gradient between the source and receiving phases under dark conditions, and the transporting ability was remarkably increased by photoirradiation. PMID- 15801731 TI - Nanogold plasmon resonance-based glucose sensing. 2. Wavelength-ratiometric resonance light scattering. AB - Gold colloids are well known to display strong plasmon absorption bands due to electron oscillations induced by the incident light. When the colloids are in proximity, the plasmon absorption bands are often perturbed. This has enabled us recently to successfully develop a glucose sensing platform based on the disassociation of dextran-coated gold colloids, cross-linked with Con A, by glucose. However, a much more useful and simpler property of gold colloids, which has been ill explored with regard to sensing, is their ability to efficiently scatter excitation light. We have found that our nanogold sensing aggregates are indeed efficient light scatters around the nanogold plasmon absorption band. By measuring the ratio of scattered light intensities at two different arbitrary wavelengths, 560 and 680 nm, glucose concentrations can be readily determined from a few millimolar up to approximately 60 mM, using a simple white light LED and detection system. Further, by measuring the ratio of the scattered intensities, this sensing approach is independent of the total sensing aggregate concentration and the excitation and detection instrumentation fluctuations or drifts. This simplistic and low-cost approach to glucose sensing, coupled with the sensing aggreagates' ability to scatter red light, suggests the potential use of these aggregates for use in physiological transdermal glucose monitoring, either for implantable skin sensors or glucose sensing tattoos (discussed later). PMID- 15801732 TI - Oligomerized phospholipid bilayers as semipermanent coatings in capillary electrophoresis. AB - Double-chained surfactants form semipermanent coatings that prevent protein adsorption in capillary electrophoresis (CE). To make such coatings more permanent, vesicles of the unsaturated phospholipid 1,2-dioleoyl-sn-glycero-3 phosphocholine were prepared and subjected to free-radical-initiated polymerization, both inside the capillary and in free solution. The latter generated oligomers of 2-5 units based on ESI-TOF MS, and formed the more stable coating in CE. Rinsing the capillary with a solution of the ex situ oligomerized DOPC suppressed EOF (0.8 x 10(-)(8) m(2)/V.s) for more than 20 h, whereas in situ oligomerized electroosmotic flow (EOF) suppressed the EOF for only 10 h. Mixtures of anionic and cationic proteins were separated under neutral pH and low ionic strength buffer with efficiencies of 480,000-930,000 plates/m and recoveries of 75-99%. PMID- 15801733 TI - Development of mild extraction methods for the analysis of natural dyes in textiles of historical interest using LC-diode array detector-MS. AB - Analysis of dyes extracted from textiles of historical interest can give valuable information as to where, when, and how the textiles were made. The most widely used method for extraction of colorants involves heating with HCl, which frequently decomposes glycosidic dye components to their parent aglycons, with consequent loss of information about the source of the dye. This is particularly true for flavonoid dyes, many of which are glycosides. We have developed or improved upon two mild textile extraction methods that use ethylenediaminetetraacetic acid (EDTA) and formic acid and are efficient in extracting dyes, but preserve glycosidic linkages. The relative efficiencies of the HCl, EDTA, and formic acid extraction methods are compared by analyzing extracts of dyed samples of silk using HPLC coupled with diode array and mass spectrometric detection. HPLC profiles of EDTA or formic acid extracts of silk dyed, for example, with pagoda tree buds and onionskins are clearly distinguishable as to the plant material used, whereas profiles of HCl extracts are not. Thus, extraction of textiles with EDTA or formic acid reagents can yield significantly more information about the original dyestuff than can extraction with a strong acid. PMID- 15801734 TI - Liquid chromatography-mass spectrometry and 15N metabolic labeling for quantitative metabolic profiling. AB - Metabolomics, i.e., the global analysis of cellular metabolites, is becoming a powerful tool for gaining insights into biological functions in the postgenomic context. However, absolute quantitation of endogenous metabolites in biological media remains an issue, and available technologies for the analysis of metabolome still lack robustness and accuracy. We describe here a new method based on liquid chromatography-mass spectrometry and (15)N uniform metabolic labeling of Saccharomyces cerevisiae for accurate and absolute quantitation of nitrogen containing cell metabolites in metabolic profiling experiments. As a proof of concept study, eight sulfur metabolites involved in the glutathione biosynthesis pathway (i.e., cysteine, homocysteine, methionine, gamma-glutamylcysteine, cystathionine, reduced and oxidized forms of glutathione, and S adenosylhomocysteine) were simultaneously quantified. The analytical method has been validated by studies of stability, selectivity, precision, and linearity and by the determination of the limits of detection and quantification. It was then applied to the analysis of extracts from cadmium-treated yeasts. In these conditions, the intracellular concentrations of most of the metabolites involved in the glutathione biosynthesis pathway were increased when compared to control extracts. These data correlate with previous proteomic results and also underline the importance of glutathione in cadmium detoxication. PMID- 15801735 TI - Determination of fractional synthesis rates of mouse hepatic proteins via metabolic 13C-labeling, MALDI-TOF MS and analysis of relative isotopologue abundances using average masses. AB - Proteins of a liver extract taken from a metabolically (13)C-labeled mouse were separated by 2D-PAGE and identified after tryptic digestion by MALDI-TOF MS peptide mass fingerprinting. (13)C-Labeling of proteins was achieved by an infusion of U-(13)C-glucose, which is metabolized to labeled nonessential amino acids. The labeling was analyzed using the relative isotopologue abundances of the measured isotope pattern of tryptic peptides and quantified by their increase in the average molecular mass (DeltaAVM). Fractional synthesis rates (FSR) of proteins were determined from corresponding peptides using measured DeltaAVM values as well as DeltaAVM values deduced from tRNA-precursor amino acid labeling, which in turn was derived from proteins showing high (13)C enrichments. The 8-h FSR values of 43 proteins were determined to range from 0 +/- 0.6 to 95 +/- 1%/8 h, with typical errors given as SEM values, which depend on the number of peptides of a specific protein usable for calculation. The method demonstrates that FSR values as an indicator for protein turnover in the liver proteome can be estimated within narrow error margins, providing baseline values from which treatment-dependent deviations could be detected with high statistical certainty. PMID- 15801736 TI - Real-time protein kinase assay. AB - We report a novel, real-time fluorogenic kinase assay. The peptide substrates are synthesized with a fluorescent dye and a hydrocarbon tail. The substrate self assembles into micelles, increasing the local concentration of the dye and quenching its fluorescence. Upon phosphorylation, the fluorescence intensity increases 4-6-fold due to micelle reorganization. Both dynamic light scattering data and cryoelectron microscope images show that the size and the shape of the phosphopeptide micelles are significantly different from micelles of substrate peptide. The system provides a robust fluorescence increase in a real-time protein kinase assay. Unlike other fluorogenic systems, the fluorophore may be distant from the serine, threonine, or tyrosine that is phosphorylated. Assays for several kinases, including PKA, PKC, p38, MAPKAP K2, akt, Erk1, and src family kinases, have been developed. IC(50) values of inhibitors for PKC betaII determined with this technology are consistent with published values. The utility of this assay to high-throughput screening was demonstrated with Sigma's LOPAC library, a collection of 640 compounds with known biological activities, and satisfactory results were obtained. PMID- 15801737 TI - Identification of molecular target of AMP-activated protein kinase activator by affinity purification and mass spectrometry. AB - We show an efficient method to identify molecular targets of small molecular compounds by affinity purification and mass spectrometry. Binding proteins were isolated from target cell lysate using affinity columns, which immobilized the active and inactive compounds. All proteins bound to these affinity columns were eluted by digestion using trypsin and then were identified by mass spectrometry. The specific binding proteins to the active compound, a candidate for molecular targets, were determined by subtracting the identified proteins in an inactive compound-immobilized affinity column from that in an active compound-immobilized affinity column. This method was applied to identification of molecular targets of D942, a furancarboxylic acid derivative, which increases glucose uptake in L6 myocytes through AMP-activated protein kinase (AMPK) activation. To elucidate the mechanism of AMPK activation by D942, affinity columns that immobilized D942 and its inactive derivative, D768, were prepared, and the binding proteins were purified from L6 cell lysate. NAD(P)H dehydrogenase [quinone] 1 (complex I), which was shown as one of the specific binding proteins to D942 by subtracting the binding proteins to D768, was partially inhibited by D942, not D768. Because inhibition of complex I activity led to a decrease in the ATP/AMP ratio, and the change in the ATP/AMP ratio triggered AMPK activation, we identified complex I as a potential protein target of AMPK activation by D942. This result shows our approach can provide crucial information about the molecular targets of small molecular compounds, especially target proteins not yet identified. PMID- 15801738 TI - Quantitative measurement of DNA adducts using neutral hydrolysis and LC-MS. Validation of genotoxicity sensors. AB - Neutral hydrolysis and LC-MS/MS analysis of 6-nm-thick DNA-polyion films used in voltammetric genotoxicity screening sensors showed that concentrations of N7 guanine DNA adducts with methyl methanesulfonate and styrene oxide increased with incubation time with the same trends as found for sensor response. Results show that the genotoxicity sensors can be used to estimate relative DNA damage rates for chemical toxicity screening. Neutral thermal hydrolysis provided a relatively clean sample matrix allowing quantitative estimates of nucleobase adducts after several minutes of incubation with damage agents. In addition, an approximate standardization procedure for neutral thermal hydrolysis was developed and validated that avoids need for a pure standard and should be useful in cases where nucleobase adduct standards are unavailable or where their identities are unknown. PMID- 15801740 TI - Sheathless capillary electrophoresis/electrospray ionization mass spectrometry using a pulled bare fused-silica capillary as the electrospray emitter. AB - It has always been assumed that electrical contact at the capillary outlet is a necessary requirement when coupling capillary electrophoresis (CE) with electrospray ionization mass spectrometry (ESI-MS). In this study, we used a pulled bare-capillary tip as the ESI emitter, but neither was it coated with any electrically conductive materials nor was a high external voltage applied on its outlet. In this paper, we demonstrate that this straightforward approach may be used to generate multiply charged ions of proteins and peptides through electrospray ionization. Our results indicate that peptides and proteins, including bradykinin, cytochrome c, myoglobin, and tryptic digest products that elute from a pulled bare-capillary tip can be detected directly by ESI-MS using the tapered bare-capillary interface. Thus, we have demonstrated that CE and ESI MS may be combined successfully without the need to modify the outlet of the capillary tip with an electrically contacting material. PMID- 15801739 TI - HPLC separation of different generations of poly(amidoamine) dendrimers modified with various terminal groups. AB - Polyamidoamine (PAMAM) dendrimers of different generations with various terminal groups were analyzed, for the first time, using a combination of high-performance liquid chromatography (HPLC), size exclusion chromatography (SEC), and matrix assisted laser desorption/ionization-time-of-flight (MALDI-TOF) techniques. Separation of amine-terminated dendrimers from generation 1 through generation 9 (G1NH(2)-G9NH(2)) was achieved using reversed-phase HPLC with elution time increasing gradually as the density of terminal amine groups increases as a function of generation. Furthermore, separation of dendrimers with terminal amino, acetamide, hydroxyl, and carboxylate groups was obtained. It has also been shown that HPLC can be used to separate dendrimers based on some structural defects inherent during the syntheses of PAMAM dendrimers. MALDI-TOF mass spectra of G1NH(2) identify the major imperfections present during typical synthesis processes. The absolute molar masses (M(n)) and molar mass distributions of the dendrimers were measured using the SEC system equipped with multiangle laser light scattering and refractive-index detectors. Findings from this study suggest HPLC can be a vital tool for characterization and preparative separation of PAMAM dendrimers. PMID- 15801741 TI - Stop-flow programmable selectivity with a dual-column ensemble of microfabricated etched silicon columns and air as carrier gas. AB - A series-coupled ensemble of microfabricated GC columns made by dry reactive ion etching of silicon substrates is evaluated for use with pneumatic selectivity enhancement techniques for targeted pairs of volatile organic compounds. Each column is 3.0 m long with a 150 miceom wide by 240 microm deep cross section. Dynamic coating was used to prepare a nonpolar column with a dimethyl polysiloxane stationary phase and a moderately polar column with a trifluoropropylmethyl polysiloxane stationary phase. Each column generates 5000 6000 theoretical plates. The columns are operated in series with the nonpolar column connected to a split inlet, the polar column connected to a flame ionization detector, and a valve connected between the column junction point and the inlet to the first column. When the valve is closed, the effluent from the first column passes directly into the second column. When the valve is open, both ends of the first column are at the inlet pressure, and flow stops in this column while increased flow is obtained in the second column. For analyte pairs that are separated by the first column but coelute from the column ensemble, the valve is opened for a few seconds after the first component of the pair has passed into the second column but the second component is still in the first column. The result is enhanced separation of the pair in the ensemble chromatogram. Relatively thick cross-linked stationary-phase films are used to increase retention for volatile compounds. The combination of air carrier gas and stationary-phase film thickness in the range 1-2 microm requires the use of relatively low average carrier gas velocities (typically less than 10 cm/s) for adequate resolving power of the column ensemble. Selectivity enhancement under isothermal conditions for a 14-component mixture of volatile organic compounds is demonstrated where neither of the columns alone nor the column ensemble without selectivity enhancement could obtain a complete separation. PMID- 15801742 TI - Coumarin tags for analysis of peptides by MALDI-TOF MS and MS/MS. 2. Alexa Fluor 350 tag for increased peptide and protein Identification by LC-MALDI-TOF/TOF MS. AB - The goal of this study was the development of N-terminal tags to improve peptide identification using high-throughput MALDI-TOF/TOF MS. Part 1 of the study was focused on the influence of derivatization on the intensities of MALDI-TOF MS signals of peptides. In part 2, various derivatization approaches for the improvement of peptide fragmentation efficiency in MALDI-TOF/TOF MS are explored. We demonstrate that permanent cation tags, while significantly improving signal intensity in the MS mode, lead to severe suppression of MS/MS fragmentation, making these tags unsuitable for high-throughput MALDI-TOF/TOF MS analysis. In the present work, it was found that labeling with Alexa Fluor 350, a coumarin tag containing a sulfo group, along with guanidation of epsilon-amino groups of Lys, could enhance unimolecular fragmentation of peptides with the formation of a high intensity y-ion series, while the peptide intensities in the MS mode were not severely affected. LC-MALDI-TOF/TOF MS analysis of tryptic peptides from the SCX fractions of an E. coli lysate revealed improved peptide scores, a doubling of the total number of peptides, and a 30% increase in the number of proteins identified, as a result of labeling. Furthermore, by combining the data from native and labeled samples, confidence in correct identification was increased, as many proteins were identified by different peptides in the native and labeled data sets. Additionally, derivatization was found not to impair chromatographic behavior of peptides. All these factors suggest that labeling with Alexa Fluor 350 is a promising approach to the high-throughput LC-MALDI-TOF/TOF MS analysis of proteomic samples. PMID- 15801743 TI - Simultaneous analysis of 2-aminopyridine-derivatized neutral and sialylated oligosaccharides from human serum in the negative-ion mode by sonic spray ionization ion trap mass spectrometry. AB - Neutral and acidic (sialylated) 2-aminopyridine-derivatized (PA) oligosaccharides were analyzed by using reversed-phase high-performance liquid chromatography/ion trap mass spectrometry (RP-HPLC/IT MS) with a sonic spray ionization (SSI) source. Under the RP-HPLC separation using a buffer of 1 mM ammonium acetate (pH4.3) at a flow rate of 0.2 mL/min, both PA-oligosaccharides in the negative ion mode showed a comparable degree of ionization efficiency, differing from that of the positive-ion mode, which exhibits a wide gap between their ionization efficiencies. In addition, the ion intensities of both PA-oligosaccharides were higher in the negative-ion mode than in the positive-ion mode. These results strongly suggest that the negative-ion mode of SSI-MS is suitable for simultaneous analysis of neutral and acidic (sialylated) oligosaccharides in RP HPLC/MS. In the present study, RP-HPLC/SSI-IT MS in the negative-ion mode was used in the analysis of PA-oligosaccharides from human serum and its usefulness was investigated. As a result, 32 neutral and sialylated PA-oligosaccharides from human serum were identified with differentiating isomeric oligosaccharides and relatively quantified by a single HPLC/MS run. This method is useful for simple and rapid analysis of the overall distribution of neutral and sialylated oligosaccharides in a complex sample such as serum. PMID- 15801744 TI - Metal salts for molecular ion yield enhancement in organic secondary ion mass spectrometry: a critical assessment. AB - In a search for molecular ion signal enhancement in organic SIMS, the efficiency of a series of organic and inorganic salts for molecular cationization has been tested using a panel of nonvolatile molecules with very different chemical characteristics (leucine enkephalin, Irganox 1010, tetraphenylnaphthalene, polystyrene). The compounds used for cationization include alkali bromide and group Ib metal salts (XBr with X = Li, Na, K; CF3CO2Ag; AgNO3; [CH3COCH=C(O )CH3]2Cu; AuCl3). Alkali ions, very good for polar molecule cationization, prove to be of limited interest for nonpolar molecules such as polystyrene. Silver trifluoroacetate displays excellent results for all the considered molecules, except for leucine enkephalin (which might be due to the use of different solvents for the analyte and the salt). Instead, silver nitrate mixed with leucine enkephalin in an ethanol solution provides intense molecular signals. The influence of the respective concentrations of analyte and salt in solution, of the silver trifluoroacetate solution stability, and of the sample microstructure on the secondary ion intensities are also investigated. The results of other combinations of analyte and salts are reported. Finally, the use of salts is critically compared to other sample preparation procedures previously proposed for SIMS analysis of large organic molecules. PMID- 15801745 TI - Simulation of evolution-selected propeptide by high-throughput selection of a peptidomimetic inhibitor on a capillary DNA sequencer platform. AB - Many proteinases, including gelatinase B/MMP-9, fulfill crucial regulatory or effector functions in disease states and may be pharmacologically targeted by specific inhibitors. Denatured collagen type II provides one of the best gelatinase B substrates, and the characteristics of its cleavage were employed to define the requirements of a novel optimal substrate probe. A synthetic fluorescent derivative was used for the development of a new high-throughput technology for the selection of inhibitors on the principles of sensitivity of confocal fluorescence detection, resolution capacity of capillary electrophoresis, and multichannel power of DNA sequencers. Combinatorial chemical synthesis of a library of peptide-based inhibitors, library deconvolution, high throughput screening, isolation, and mass spectrometric techniques enabled us to identify a novel single-peptide gelatinase B inhibitor. A notable finding is that the in vitro-selected inhibitor mimics many of the characteristics of the evolution-selected MMP propeptide sequence. PMID- 15801746 TI - Development of a microchip-based bioassay system using cultured cells. AB - We developed a novel bioassay system using a glass microchip and cultured cells. A microchamber for cell culture and microchannels for reactions and detection were fabricated on a Pyrex glass substrate by photolithography and wet etching techniques. Cell culture, chemical and enzymatic reactions, and detection were integrated into the microchip. To keep different temperatures locally in three areas of the microchip, we designed and fabricated a temperature control device. Nitric oxide released from macrophage-like cells stimulated by lipopolysaccharide was successfully monitored with the microchip, the temperature control device, and a thermal lens microscope. The total assay time was reduced from 24 to 4 h, and detection limit of NO was improved from 1 x 10(-6) to 7 x 10(-8) M compared with conventional methods. Moreover, the system could monitor a time course of the release, which is difficult to measure by conventional batch methods. We conclude that this system is promising for a rapid bioassay system with very small consumption of cells. PMID- 15801747 TI - Spectroelectrochemical microscopy: spatially resolved spectroelectrochemistry of carrier-based ion-selective membranes. AB - High-resolution spectroscopic imaging of the cross section of ion-selective membranes and the adjoining solution phases during real-time electrochemical measurement is termed as spectroelectrochemical microscopy (SpECM). The novel SpECM instrument utilizes wavelength-dispersive multispectral imaging of a thin membrane strip separating the two sides of a four-electrode thin-layer electrochemical cell. SpECM is aimed as a tool for optimizing the experimental conditions in mass transport-controlled ion-selective electrode membranes for improved detection limit. Some of the capabilities of the new technique are demonstrated using fix site, chromoionophore-based, pH-sensitive membranes as model systems. The experimental results are discussed in the light of the existing theory of fixed-site membranes. The quantitative expression for the time dependent change of the free ionophore concentration across the ion-selective membrane showed close correlation to the recorded concentration profiles. PMID- 15801748 TI - High-performance genetic analysis on microfabricated capillary array electrophoresis plastic chips fabricated by injection molding. AB - We have developed a novel technique for mass production of microfabricated capillary array electrophoresis (mu-CAE) plastic chips for high-speed, high throughput genetic analysis. The mu-CAE chips, containing 10 individual separation channels of 50-microm width, 50-microm depth, and a 100-microm lane-to lane spacing at the detection region and a sacrificial channel network, were fabricated on a poly(methyl methacrylate) substrate by injection molding and then bonded manually using a pressure-sensitive sealing tape within several seconds at room temperature. The conditions for injection molding and bonding were carefully characterized to yield mu-CAE chips with well-defined channel and injection structures. A CCD camera equipped with an image intensifier was used to monitor simultaneously the separation in a 10-channel array with laser-induced fluorescence detection. High-performance electrophoretic separations of phiX174 HaeIII DNA restriction fragments and PCR products related to the human beta globin gene and SP-B gene (the surfactant protein B) have been demonstrated on mu CAE plastic chips using a methylcellulose sieving matrix in individual channels. The current work demonstrated greatly simplified the fabrication process as well as a detection scheme for mu-CAE chips and will bring the low-cost mass production and application of mu-CAE plastic chips for genetic analysis. PMID- 15801749 TI - Permeability of the nuclear envelope at isolated Xenopus oocyte nuclei studied by scanning electrochemical microscopy. AB - In interphase eukaryotic cells, molecular transport between the cytoplasm and the nucleus is mediated by the nuclear pore complex (NPC), which perforates the double-membraned nuclear envelope (NE). Local permeability of the NE at large intact nuclei (approximately 400 microm in diameter) isolated from Xenopus laevis oocytes was studied by scanning electrochemical microscopy (SECM). Steady-state tip current versus tip-nucleus distance curves (approach curves) were measured with 10- and 2-microm-diameter Pt disk microelectrodes at the nuclei in isotonic buffer solutions containing redox-active molecules. The approach curves in the normalized form are independent of the tip diameter, indicating diffusion-limited membrane transport of the redox molecules. SECM chronoamperometry demonstrated that a decrease in the steady-state tip current at short tip-nucleus distances is due to smaller diffusion coefficients and concentrations of the redox molecules in the nucleus than those in the buffer solution. The experimental approach curves fit very well with theoretical ones for freely permeable membranes, yielding the NE permeability to the molecules that is at least 2 orders of magnitude larger than permeability of bilayer lipid membranes and cell membranes. This result indicates that passive transport of the redox molecules across the NE is facilitated by open NPC pores. The flux of the redox molecules sustainable by a single NPC channel (>9.8 x 10(6) molecules per NPC per second) and the diameter of the channel pore (>15 nm) were estimated from the SECM data by assuming the NE as an array of nanometer-sized NPC pores. The effects of the redox molecules on the nucleus and the NPC function were examined by studying signal-mediated nuclear import of rhodamine-labeled bovine serum albumin with and without nuclear localization signals by fluorescence microscopy. PMID- 15801750 TI - Electrostatic contributions to protein retention in ion-exchange chromatography. 2. Proteins with various degrees of structural differences. AB - The relation of protein structure to retention provides a framework within which to investigate chromatographic adsorption mechanisms. Protein sets with varying degrees of structural differences were studied to relate variations in protein properties to retention behavior. To explore molecular contributions to protein adsorption in ion-exchange chromatography, protein-adsorbent electrostatic interactions were modeled using a continuum approach. The calculations qualitatively capture the chromatographic differentiation of closely related subtilisin variants. Descriptions of the electrostatic interactions of FGF-1 vs FGF-2 with cation exchangers were obtained, and aid in rationalizing differences in experimental retention trends across a set of adsorbents based on different adsorption mechanisms linked to the adsorbent structure. Comparative calculations for proteins with differences in local or overall arginine-lysine composition, including subtilisin variants G166R/G166K and lysozyme/cytochrome c, suggest that continuum electrostatics is not adequate to capture the full quantitative characteristics of the chromatographic retention of proteins. To allow more accurate description of retention, additional molecular interactions, specifically hydration effects, must be incorporated in the model. PMID- 15801751 TI - Automated quantitative analysis of complex lipidomes by liquid chromatography/mass spectrometry. AB - Recent advances in mass spectrometry have revolutionized the analysis of lipid compositions of cells and other biomaterials by simplifying the analytical protocol dramatically and by increasing the sensitivity of detection by several orders of magnitude. However, the throughput of the published mass spectrometric methods is severely limited by data analysis, which requires extensive operator involvement. Consequently, we have developed an automated method that allows unattended identification and quantification of lipid molecular species of all the major lipid classes from a two-dimensional chromatographic/mass spectrometric data set. More than 100 polar lipid species could be automatically quantified from different biological samples with good accuracy and reproducibility. The response was linear over approximately 3 orders of magnitude with the equipment used, and approximately 35 samples could be analyzed in a day. This method makes high-throughput lipidomics feasible in biology, biotechnology, and medicine. PMID- 15801752 TI - Improvement in peptide detection for proteomics analyses using NanoLC-MS and high field asymmetry waveform ion mobility mass spectrometry. AB - Sensitive and selective detection of multiply charged peptide ions from complex tryptic digests was achieved using high-field asymmetric waveform ion mobility spectrometry (FAIMS) combined with nanoscale liquid chromatography-mass spectrometry (nanoLC-FAIMS-MS). The combination of FAIMS provided a marked advantage over conventional nanoLC-MS experiments by reducing the extent of chemical noise associated with singly charged ions and enhancing the overall population of detectable tryptic peptides. Such advantages were evidenced by a 6 12-fold improvement in signal-to-noise ratio measurements for a wide range of multiply charged peptide ions. An increase of 20% in the number of detected peptides compared to conventional nanoelectrospray was achieved by transmitting ions of different mobilities at high electric field vs low field while simultaneously recording each ion population in separate mass spectrometry acquisition channels. This method provided excellent reproducibility across replicate nanoLC-FAIMS-MS runs with more than 90% of all detected peptide ions showing less than 30% variation in intensity. The application of this technique in the context of proteomics research is demonstrated for the identification of trace-level proteins showing differential expression in U937 monocyte cell extracts following incubation with phorbol ester. PMID- 15801753 TI - Quantitative proteomic analysis by accurate mass retention time pairs. AB - Current methodologies for protein quantitation include 2-dimensional gel electrophoresis techniques, metabolic labeling, and stable isotope labeling methods to name only a few. The current literature illustrates both pros and cons for each of the previously mentioned methodologies. Keeping with the teachings of William of Ockham, "with all things being equal the simplest solution tends to be correct", a simple LC/MS based methodology is presented that allows relative changes in abundance of proteins in highly complex mixtures to be determined. Utilizing a reproducible chromatographic separations system along with the high mass resolution and mass accuracy of an orthogonal time-of-flight mass spectrometer, the quantitative comparison of tens of thousands of ions emanating from identically prepared control and experimental samples can be made. Using this configuration, we can determine the change in relative abundance of a small number of ions between the two conditions solely by accurate mass and retention time. Employing standard operating procedures for both sample preparation and ESI mass spectrometry, one typically obtains under 5 ppm mass precision and quantitative variations between 10 and 15%. The principal focus of this paper will demonstrate the quantitative aspects of the methodology and continue with a discussion of the associated, complementary qualitative capabilities. PMID- 15801754 TI - Metabolomic analysis of eukaryotic tissue and prokaryotes using negative mode MALDI time-of-flight mass spectrometry. AB - Metabolites in islets of Langerhans and Escherichia coli strain DH5-alpha were analyzed using negative-mode, matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF-MS). For analysis of anionic metabolites by MALDI, 9-aminoacridine as the matrix yielded a far superior signal in comparison to alpha-cyano-4-hydroxycinnamic acid, 2,5-dihydrobenzoic acid, 2,4,6, trihydroxyacetophenone, and 3-hydroxypicolinic acid. Limits of detection for metabolite standards were as low as 15 nM for GDP, GTP, ADP, and ATP and as high as 1 muM for succinate in 1-muL samples. Analysis of islet extracts allowed detection of 44 metabolites, 29 of which were tentatively identified by matching molecular weight to compounds in METLIN and KEGG databases. Relative quantification was demonstrated by comparing the ratio of selected di- and triphosphorylated nucleotides for islets incubated with different concentrations of glucose. For islets at 3 mM glucose, concentration ratios of ATP/ADP, GTP/GDP, and UTP/UDP were 1.9 +/- 1.39, 1.12 +/- 0.50, and 0.79 +/- 0.35 respectively, and at 20 mM glucose stimulation, the ratios increased to 4.13 +/- 1.89, 5.62 +/ 4.48, and 4.30 +/- 4.07 (n = 3). Analysis was also performed by placing individual, intact islets on a MALDI target plate with matrix and impinging the laser directly on the dried islet. Direct analysis of single islets allowed detection of 43 metabolites, 28 of which were database identifiable. A total of 43% of detected metabolites from direct islet analysis were different from those detected in islet extracts. The method was extended to prokaryotic cells by analysis of extracts from E. coli. Sixty metabolites were detected, 39 of which matched compounds in the MetaCyc database. A total of 27% of the metabolites detected from prokaryotes overlapped those found in islets. These results show that MALDI can be used for detection of metabolites in complex biological samples. PMID- 15801755 TI - Characterization and matching of oil samples using fluorescence spectroscopy and parallel factor analysis. AB - A novel approach for matching oil samples by fluorescence spectroscopy combined with three-way decomposition of spectra is presented. It offers an objective fingerprinting based on the relative composition of polycyclic aromatic compounds (PACs) in oils. The method is complementary to GC-FID for initial screening of oil samples but can also be used for prescreening in the field, onboard ships, using a portable fluorescence spectrometer. Parallel factor analysis (PARAFAC) was applied to fluorescence excitation-emission matrixes (EEMs) of heavy fuel oils (HFOs), light fuel oils, lubricating oils, crude oils, unknown oils, and a sample collected in the spill area two weeks after the Baltic Carrier oil spill (Denmark, 2001). A total of 112 EEMs were decomposed into a five-factor PARAFAC model using excitation wavelengths from 245 to 400 nm and emission wavelengths from 280 to 550 nm. The PARAFAC factors were compared to EEMs of PAC standards with two to five rings, and the comparisons indicate that each of the factors can be related to a mixture of PACs with similar fluorescence characteristics: a mixture of naphthalenes and dibenzothiophenes, fluorenes, phenanthrenes, chrysenes, and five-ring PACs, respectively. Oils were grouped in score plots according to oil type. Except for HFOs and crude oils, the method easily discriminated between the four oil types. Minor overlaps of HFOs and crude oils were observed along all five PARAFAC factors, and the variability of crude oils was large along factor 2 due to a varying content of five-ring PACs. The spill sample was correctly assigned as a HFO with similar PAC pattern as oil from the cargo tank of the Baltic Carrier by comparing the correlation coefficient of scores for the oil spill sample and possible source oils (i.e., oils in the database). PMID- 15801756 TI - Single-photon ionization quadrupole mass spectrometry with an electron beam pumped excimer light source. AB - The application of soft ionization methods for mass spectrometry (MS), such as single-photon ionization (SPI) using vacuum ultraviolet (VUV) light, provides powerful analytical instrumentation for real-time on-line monitoring of organic substances in gaseous matrixes. A compact and mobile quadrupole mass spectrometer (QMS) system using a novel electron beam pumped rare gas VUV lamp for SPI has been developed for on-line analysis of organic trace compounds (ppb concentrations). The VUV radiation of the light source is employed for SPI in the ion source of the QMS. The concept of the interfacing of the VUV light source with the QMS is described and the SPI-QMS is characterized. On-line detection limits down to 50 ppb for benzene, toluene, and m-xylene were achieved. The instrument is well suited for continuous measurements of aromatic and aliphatic trace compounds and can therefore be used for on-line monitoring of trace compounds in dynamically fluctuating process gases. First measurements of gas standards, petrochemical samples, and on-line monitoring of automotive exhaust are presented. PMID- 15801757 TI - Tackling calibration problems of spectroscopic analysis in high-throughput experimentation. AB - High-throughput experimentation and screening methods are changing work flows and creating new possibilities in biochemistry, organometallic chemistry, and catalysis. However, many high-throughput systems rely on off-line chromatography methods that shift the bottleneck to the analysis stage. On-line or at-line spectroscopic analysis is an attractive alternative. It is fast, noninvasive, and nondestructive and requires no sample handling. The disadvantage is that spectroscopic calibration is time-consuming and complex. Ideally, the calibration model should give reliable predictions while keeping the number of calibration samples to a minimum. In this paper, we employ the net analyte signal approach to build a calibration model for Fourier transform near-infrared measurements, using a minimum number of calibration samples based on blank samples. This approach fits very well to high-throughput setups. With this approach, we can reduce the number of calibration samples to the number of chemical components in the system. Thus, the question is no longer how many but which type of calibration samples should one include in the model to obtain reliable predictions. Various calibration models are tested using Monte Carlo simulations, and the results are compared with experimental data for palladium-catalyzed Heck cross-coupling. PMID- 15801758 TI - Comparison of photophysical and colloidal properties of biocompatible semiconductor nanocrystals using fluorescence correlation spectroscopy. AB - A number of different surface chemistries have been developed in recent years to render semiconductor nanocrystals (NCs) stable in water and biocompatible. However, most of these surface modifications affect NCs' photophysical properties, calling for a method to simultaneously monitor colloidal and fluorescence properties. Fluorescence correlation spectroscopy (FCS) combined with ensemble spectroscopic methods and Monte Carlo simulations were used to interpret and derive photophysical as well as colloidal properties of four different NC surface treatments. Using a novel FCS scheme with alternating laser excitation at two different intensities, we first ruled out influences from optical gradient forces (optical trapping). We then compared concentration of emitting particles, brightness per particle, saturation intensity, blinking (intermittency), hydrodynamic radius, and propensity for aggregation of the different bioconjugated NCs. This approach was successfully applied during the development and optimization of peptide-coated NCs. PMID- 15801759 TI - Data analysis for a dual-channel virus counter. AB - A simple algorithm is presented for quantitative analysis of simultaneous events on a dual-channel flow cytometer designed specifically for virus counting. The algorithm, which is based on matrix analysis of burst lag times, was evaluated using baculovirus samples that had previously been quantified by the plaque titer method. The results indicated statistical reliability for the algorithm, with three of six samples yielding the same value, within error, for viruses per unit volume as the plaque titer. The other three samples yielded values within a factor approximately 2, which was deemed acceptable given the limitations of the plaque titer method. PMID- 15801760 TI - Resin bead micro-UV-visible absorption spectroscopy. AB - The construction and design of a microscope coupled with a miniature UV-vis spectrometer is described. This was applied to the study of dyes linked to solid supports and displayed good correlation in spectral shape and lambda(max) values when compared to the dyes in solution, as well as showing a linear relationship between dye loading and UV-vis absorbance. The spectral profiles of these dyes at various pH's were measured and used to determine the pK(a) of the dyes on the beads, which were compared with the pK(a) values of the dyes in solution, thus enabling the dye-loaded beads to act as pH sensors. PMID- 15801761 TI - Polymer nanochannels fabricated by thermomechanical deformation for single molecule analysis. AB - A simple method for fabricating nanoscale channels based on thermomechanical deformation of rigid polymer substrates is demonstrated. Polycarbonate preforms containing microchannels with cross-sectional dimensions on the order of tens of micrometers are controllably deformed to produce submicrometer dimensions. The reduced channel dimensions are achieved by heating the preform while applying a uniaxial tensile force to reduce channel cross sections through the Poisson effect. Nanochannels with circular or elliptical cross sections are defined by varying the channel position and preform geometry prior to deformation. Arrays of parallel nanochannels with critical dimensions down to 400 nm are described. Using the fabrication method, a nanochannel network is fabricated for the detection of single protein molecules via confocal fluorescence microscopy. The chip includes a detection channel with cross-sectional dimensions approaching the confocal volume dimensions of the detection optics and a larger adjacent reference channel used to optimize focusing. Detection of fluorescently labeled bovine serum albumin at 15 and 150 nM concentrations is presented, demonstrating the ability to perform single-molecule fluorescence measurements in polycarbonate chips using visible wavelengths for excitation and detection. PMID- 15801762 TI - Selective extraction of trace levels of polychlorinated and polybrominated contaminants by supercritical fluid-solid-phase microextraction and determination by gas chromatography/mass spectrometry. Application to aquaculture fish feed and cultured marine species. AB - The persistence, ubiquity, and toxicity of polyhalogenated compounds, together with their presence in fish feed, make it necessary to monitor these organic pollutants in the routine quality assurance programs of aquaculture activities, as this food chain is a source of these toxic compounds for human consumers. A new approach based on simultaneous supercritical fluid extraction-sample cleanup, followed by solid-phase microextraction-gas chromatography/mass spectrometry (SFE SPME-GC/MS/MS) has been developed as an advantageous analytical tool for the determination of 15 organohalogenated compounds (including pesticides, polychlorinated and polybrominated biphenyls, and polybrominated diphenyl ethers) in aquaculture feed at very low levels. The influence of several parameters in the efficiency of the SPE/SPME combination was systematically investigated by chemometric approaches. In the optimal conditions, the developed procedure provides an excellent linearity, detection, and quantification limits (below 10 pg/g) for most of the analytes investigated, being at the same time advantageous in terms of rapidity, convenience, and avoiding the need of toxic organic solvents. The procedure was applied to the analysis of aquaculture feed and cultured marine species and tested for accuracy against IAEA 406 reference material. PMID- 15801763 TI - Integration of valving and sensing on a capillary-assembled microchip. AB - A simple integration of both flow control valves and a reaction-based sensing function on a single microchip was performed by using capillary-assembled microchip (CAs-CHIP: Hisamoto, H.; Nakashima, Y.; Kitamura, C.; Funano, S.-i.; Yasuoka, M.; Morishima, K.; Kikutani, Y.; Kitamori, T.; Terabe, S. Anal. Chem. 2004, 76, 3222-3228.). In contrast to the previously reported on-chip valving systems, where the simple valving functions were integrated, our system can integrate not only valving function but also many other chemical functions to perform a complex chemical operation on a single microchip. Here, an enzymatic reaction-based readout system is employed as an example. A square capillary immobilizing N-isopropylacrylamide polymer monolith (referred to as "valving capillary") is used as a thermoresponsive "valving part" and the immobilizing enzyme-modified glycidyl methacrylate polymer monolith (referred to as "sensing capillary") is used as a "sensing part" of the CAs-CHIP. These capillaries are embedded into a lattice microchannel network fabricated on poly(dimethylsiloxane), which has the same channel dimensions as the outer dimensions of the square capillaries. After bonding, a small Peltier device (2 mm x 2 mm) for temperature control is placed on the embedded valving capillaries to control fluid flow. Using this for heating or cooling, fast operation times of 1.4 and 3.2 s for opening and closing valves, respectively, are successfully achieved. Finally, two valving capillaries are independently controlled to trap sample solution within a bypass channel, where the enzyme-immobilized capillary is embedded, and then enzymatic reaction-based sensing of chemical species is performed as an example. The fundamental characteristics of the valve-integrated microchip are fully investigated, and an application to the analysis of an enzyme substrate by using two independent valving capillaries and a sensing capillary is demonstrated. PMID- 15801764 TI - Direct determination of peracetic acid, hydrogen peroxide, and acetic acid in disinfectant solutions by far-ultraviolet absorption spectroscopy. AB - In this paper we propose a rapid and highly selective far-ultraviolet (FUV) spectroscopic method for the simultaneous determination of peracetic acid (PAA), hydrogen peroxide, and acetic acid (AA). For this purpose we developed a novel FUV spectrometer that enables us to measure the spectra down to 180 nm. Direct determination of PAA, H(2)O(2), and AA, the three main species in disinfectant solutions, was carried out by using their absorption bands in the 180-220-nm region. The proposed method does not require any reagents or catalysts, a calibration standard, and a complicated procedure for the analysis. The only preparation procedure requested is a dilution of H(2)O(2) with pure water to a concentration range lower than 0.2 wt % in the sample solutions. Usually, the required concentration range can be obtained by the 10 times volume dilution of the actual disinfectant solutions. As the measured sample does not leave any impurity for the disinfection, it can be reused completely by using a circulation system. The detection limit for PAA of the new FUV spectrometer was evaluated to be 0.002 wt %, and the dynamic ranges of the measured concentrations were from 0 to 0.05 wt %, from 0 to 0.2 wt %, and from 0 to 0.2 wt % for PAA, H(2)O(2), and AA, respectively. The response time for the simultaneous determination of the three species is 30 s, and the analysis is applicable even to the flowing samples. This method may become a novel approach for the continuous monitoring of PAA in disinfectant solutions on the process of sterilization. PMID- 15801766 TI - A cyclical semicontinuous process for production of human alpha 1-antitrypsin using metabolically induced plant cell suspension cultures. AB - Transgenic rice suspension cultures were utilized to produce a human therapeutic protein, recombinant alpha(1)-antitrypsin (rAAT), in a cyclical, semicontinuous operation. Recombinant protein production was induced by removing the carbon source from the cell culture medium. The transgenic rice cells secreted the rAAT into the medium, and therefore medium exchanges could be performed for consecutive growth and protein expression phases. The process consisted of three cycles over a 25-28 day period, with growth phases lasting 4-6 days each and protein expression phases lasting 2.5-5 days each. Biomass and sugar concentrations, oxygen uptake rate, cell viability, culture pH, total extracellular protein, and active rAAT were measured throughout the cyclical process. The data profiles were reproducible between separate cyclical runs where, following each induction period, cell growth and viability could be reestablished once sucrose was added back to the culture. Volumetric productivities ranged from 3 to 12 mg active rAAT/(L day) for individual cycles with overall volumetric productivities of 4.5 and 7.7 mg active rAAT/(L day). PMID- 15801767 TI - Design of metabolic engineering strategies for maximizing L-(-)-carnitine production by Escherichia coli. Integration of the metabolic and bioreactor levels. AB - In this work metabolic engineering strategies for maximizing L-(-)-carnitine production by Escherichia coli based on the Biochemical System Theory and the Indirect Optimization Method are presented. The model integrates the metabolic and the bioreactor levels using power-law formalism. Based on the S-system model, the Indirect Optimization Method was applied, leading to profiles of parameter values that are compatible with both the physiology of the cells and the bioreactor system operating conditions. This guarantees their viability and fitness and yields higher rates of L-(-)-carnitine production. Experimental results using a high cell density reactor were compared with optimized predictions from the Indirect Optimization Method. When two parameters (the dilution rate and the initial crotonobetaine concentration) were directly changed in the real experimental system to the prescribed optimum values, the system showed better performance in L-(-)-carnitine production (74% increase in production rate), in close agreement with the model's predictions. The model shows control points at macroscopic (reactor operation) and microscopic (molecular) levels where conversion and productivity can be increased. In accordance with the optimized solution, the next logical step to improve the L-( )-carnitine production rate will involve metabolic engineering of the E. coli strain by overexpressing the carnitine transferase, CaiB, activity and the protein carrier, CaiT, responsible for substrate and product transport in and out of the cell. By this means it is predicted production may be enhanced by up to three times the original value. PMID- 15801768 TI - Candida rugosa lipase-catalyzed intramolecular O- to N- transacylation of butyryl propranolol in the presence of cyclodextrins. AB - In the present paper, a novel enzymatic reaction between (R,S)-O-butyryl propranolol (O-BP) and lipase from Candida rugosa in the presence of hydroxypropyl-beta-cyclodextrin (HP-beta-CD) is described. Under the used condition, lipase catalyzed the intramolecular transacylation of O-BP into N butyryl propranolol (N-BP). Propranolol, the product of the expected hydrolysis reaction, was not detected in the reaction medium. A chiral analysis of the reaction product indicated that lipase showed a preference for (R)-O-butyryl propranolol since it first transformed the (R)-enantiomer and then the corresponding (S)-enantiomer. The influence of different reaction conditions on the initial rate is also studied. PMID- 15801769 TI - Auxotrophic markers pyrF and proC can replace antibiotic markers on protein production plasmids in high-cell-density Pseudomonas fluorescens fermentation. AB - The use of antibiotic-resistance genes as selectable markers in transgenic organisms is coming under increased scrutiny, for fear that they may spread to human pathogens, thereby reducing the effectiveness of antibiotic therapy. A current Pseudomonas fluorescens protein expression system uses a tetracycline resistance gene (tetR/tetA) to maintain an expression plasmid under control of a repressible promoter and a kanamycin resistance gene (kanR) to maintain a plasmid carrying a repressor gene. We investigated using auxotrophic markers to replace these two antibiotic resistance genes: pyrF (encoding orotidine-5'-phosphate decarboxylase) in place of tetR/tetA and proC (encoding pyrroline-5-carboxylate reductase) in place of kanR, complementing their respective precise chromosomal deletions created by allele exchange using a suicide vector carrying pyrF as a counterselectable marker. The resulting strains, devoid of antibiotic-resistance genes, were shown to achieve high productivity of nitrilase and thermostable alpha-amylase equal to that of the former antibiotic-resistant production host. The production plasmids were stable. The pyrF (uracil-dependent) background of the production host strain also allows us to sequentially alter the genome to incorporate other desired genomic changes, deletions, or insertions using 5' fluoroorotic acid counterselection, restoring the selectable marker after each step. PMID- 15801770 TI - Secretion and surface display of green fluorescent protein using the yeast Saccharomyces cerevisiae. AB - Green fluorescent protein (GFP) continues to be a very useful tool in biotechnology, but soluble production of GFP and GFP-protein fusions has been difficult. In this study, we have produced yeast-enhanced green fluorescent protein (yEGFP) in Saccharomyces cerevisiae as a soluble, secreted product with a purified level of 6 mg/L. Expression was directed by the inducible GAL1-10 promoter and synthetic prepro leader sequence. The secretion of yEGFP by yeast was strongly dependent on temperature, with 20 degrees C induction being optimal. Use of 2 micro multicopy expression constructs elevated yields over a low-copy CEN-based system by approximately 2-fold. Yeast-enhanced GFP was also expressed as a fusion to the Aga2p mating agglutinin in order to test the secretory processing fidelity of yEGFP-protein fusions. When the cell surface anchoring protein, Aga1p, was co-overexpressed with the Aga2p-yEGFP fusion, the Aga2p-yEGFP protein was tethered to the yeast cell surface. Flow cytometry and fluorescence microscopy analysis indicated that the fusion was displayed on the yeast cell surface at high levels. In the absence of high level Aga1p expression, the Aga2p yEGFP fusion protein was instead secreted in its entirety with no detectable surface display. These findings reveal that yeast is a suitable host for secretion of GFP and GFP-protein fusions and thus could enable a wide range of biochemistry and biotechnology applications. PMID- 15801771 TI - Efficient succinic acid production from glucose through overexpression of pyruvate carboxylase in an Escherichia coli alcohol dehydrogenase and lactate dehydrogenase mutant. AB - An adhE, ldhA double mutant Escherichia coli strain, SBS110MG, has been constructed to produce succinic acid in the presence of heterologous pyruvate carboxylase (PYC). The strategic design aims at diverting maximum quantities of NADH for succinate synthesis by inactivation of NADH competing pathways to increase succinate yield and productivity. Additionally an operational PFL enzyme allows formation of acetyl-CoA for biosynthesis and formate as a potential source of reducing equivalents. Furthermore, PYC diverts pyruvate toward OAA to favor succinate generation. SBS110MG harboring plasmid pHL413, which encodes the heterologous pyruvate carboxylase from Lactococcus lactis, produced 15.6 g/L (132 mM) of succinate from 18.7 g/L (104 mM) of glucose after 24 h of culture in an atmosphere of CO(2) yielding 1.3 mol of succinate per mole of glucose. This molar yield exceeded the maximum theoretical yield of succinate that can be achieved from glucose (1 mol/mol) under anaerobic conditions in terms of NADH balance. The current work further explores the importance of the presence of formate as a source of reducing equivalents in SBS110MG(pHL413). Inactivation of the native formate dehydrogenase pathway (FDH) in this strain significantly reduced succinate yield, suggesting that reducing power was lost in the form of formate. Additionally we investigated the effect of ptsG inactivation in SBS110MG(pHL413) to evaluate the possibility of a further increase in succinate yield. Elimination of the ptsG system increased the succinate yield to 1.4 mol/mol at the expense of a reduction in glucose consumption of 33%. In the presence of PYC and an efficient conversion of glucose to products, the ptsG mutation is not indispensable since PEP converted to pyruvate as a result of glucose phosphorylation by the glucose specific PTS permease EIICB(glu) can be rediverted toward OAA favoring succinate production. PMID- 15801773 TI - Retroviral transduction of adherent cells in resonant acoustic fields. AB - Ultrasound-induced cavitation has been extensively used to enhance the efficiency of nonviral-based gene delivery. Although such unique mechanical force could possibly augment the efficacy of retrovirus-mediated gene transfer, we harnessed an alternative approach, a resonant acoustic field, to facilitate the retroviral transduction rate. NIH 3T3 fibroblast cells suspended in a culture well and mixed with ecotropic retroviruses were co-treated with megahertz resonant acoustic fields (RAF). Suspended NIH 3T3 cells under RAF treatment agglomerated at acoustic nodal planes by primary radiation force within a short exposure time. These first arrived and agglomerated cells formed bands as nucleating sites for nanometer-sized ecotropic retroviruses circulated between nodal planes to attach on and thereby increased cell-virus encounters. According to the neomycin resistant colony assay, 2-fold increment of retroviral transduction rate was obtained by exposing cells and retroviruses in the RAF for 6 min in the presence of 8 microg/mL Polybrene. PMID- 15801772 TI - Stereospecific synthesis of (R)-2-hydroxy carboxylic acids using recombinant E. coli BL21 overexpressing YiaE from Escherichia coli K12 and glucose dehydrogenase from Bacillus subtilis. AB - The yiaE gene from Escherichia coli K12 was functionally expressed in E. coli BL21 using an IPTG inducible pET expression system (2.1 U/mg), and YiaE was purified to a specific activity of 18 U/mg. The purified enzyme catalyzes reduction of various aromatic and aliphatic 2-oxo carboxylic acids to the corresponding (R)-2-hydoxy carboxylic acids using NADPH. For practical applications, the problem of NADPH recycle was effectively solved by using recombinant E. coli overexpressing YiaE and glucose dehydrogenase from Bacillus subtilis in the same cell. The recombinant E. coli was used to prepare (R) phenyllactic acid and (R)-2-hydroxy-4-phenylbutanoic acid from the corresponding 2-oxo carboxylic acids (98% ee) while the alpha-carbonyl group of 2,4-dioxo-4 phenylbutyric acid was reduced regio- and stereospecifically to give (R)-2 hydroxy-4-oxo-4-phenylbutyric acid (97% ee) in quantitative yields. The cells could be recycled for 3 days at room temperature in 100 mM phosphate buffer (pH 7.0) without loss of activity, which reduced to 70% after 1 week. PMID- 15801774 TI - Fungal inoculum properties and its effect on growth and enzyme activity of Trametes versicolor in soil. AB - The effect of fungal inoculum properties on colonization of nonsterile soil by three isolates of the white-rot fungus Trametes versicolor was investigated. Fungal inoculum properties were examined in separate experiments and were fungal inoculum composition, age of fungal inoculum, concentration of the inoculum and inoculation method. The fungal inoculum composition study compared pine versus poplar sawdust as the basic carrier with varying amounts of corn grit, corn meal and starch. The age of the fungal inoculum studied ranged from 3 to 21 days. The inoculum concentration gradually increased from 0 to 50% (v/v). The study assessing inoculation method compared mixing with layering techniques. The effect of moisture conditions of soil, sawdust and sand in combination with two inoculation methods (mixing versus point source inoculation) on colonization by T. versicolor was also determined. Colonization of soil was always assessed visually and enzymatically monitoring mycelial growth, biological potential (fluorescein diacetate assay) and laccase levels. Generally, the three different assessment methods correlated (P < 0.05) with each other. A fungal inoculum based on pine sawdust supported white-rot fungal growth in soil better than a poplar sawdust basis. Colonization of soil by T. versicolor was improved by increasing the corn content of the fungal inoculum. Younger (<7 days old) fungal inoculum resulted in better soil colonization than older (>10 days). A strong correlation (P < 0.001) was observed between the amount of fungal inoculum used in the soil augmentation and white-rot fungal colonization of soil. Inoculation of the fungal inoculum into soil by mixing was preferable over application in layers or point source inoculation. Moisture level did not influence biological potential measurements, but affected mycelial growth and laccase expression. PMID- 15801775 TI - Maximization of production of secreted recombinant proteins in Pichia pastoris fed-batch fermentation. AB - Pontryagin's Maximum Principle has been applied for optimization of secreted proteins from Pichia pastoris fed-batch fermentation. The objective of this work is to maximize the total accumulated product per unit operation time under different given conditions and system constraints. To obtain optimal solutions, an automated curve-fitting software, Table Curve 2D, was employed to construct the necessary mathematical models and solve the complicated functions. In the solution processes, the end of the glycerol batch phase was defined as the initial state of the system, the end of the methanol fed-batch phase as the final state, the cell mass produced along with product accumulated as state variables, and the specific growth rate (mu) as the control variable. Initially, a relationship between the specific production rate (rho) and mu was established. Then, according to Pontryagin's Maximum Principle, the admissible range of mu and its trajectories for the optimal operations were determined. Four representative cases with different combinations of the operation time along with the initial and final states were evaluated. A close correlation was obtained between the predicted values of the model equation with the experimental results from the Pichia pastoris fed-batch fermentations producing secreted alpha-galactosidase. The approaches proposed here greatly simplify the computational processes and validate the optimization strategy as a generalized approach to maximize the yield from fed-batch fermentations. PMID- 15801776 TI - Optimization of rosmarinic acid production by Lavandula vera MM plant cell suspension in a laboratory bioreactor. AB - The all-round effect of dissolved oxygen concentration, agitation speed, and temperature on the rosmarinic acid production by Lavandula vera MM cell suspension was studied in a 3-L laboratory bioreactor by means of the modified Simplex method. Polynomial regression models were elaborated for description of the process of rosmarinic acid production (Y) in the bioreactor as a consequence of the variation of the dissolved oxygen (X(1)) concentration between 10% and 50%; agitation (X(2)) between 100 and 400 rpm; and temperature (X(3)) between 22 and 30 degrees C. The optimization made it possible to establish the optimal conditions for the biosynthesis of rosmarinic acid by L. vera MM: dissolved oxygen (X(1)*), 50% of air saturation; agitation (X(2)*), 400 rpm; and temperature (X(3)*), 29.9 degrees C, where maximal yield (Y(max)) of 3489.4 mg/L of rosmarinic acid was achieved (2 times higher compared with the shake-flasks cultivation). PMID- 15801777 TI - Lipase-catalyzed acyl exchange of soybean phosphatidylcholine in n-Hexane: a critical evaluation of both acyl incorporation and product recovery. AB - Lipase-catalyzed acidolysis was examined for the production of structured phospholipids in a hexane system. In a practical operation of the reaction system, the formation of lyso-phospholipids from hydrolysis is often a serious problem, as demonstrated from previous studies. A clear elucidation of the issue and optimization of the system are essential for the practical applications in reality. The effects of enzyme dosage, reaction temperature, solvent amount, reaction time, and substrate ratio were optimized in terms of the acyl incorporation, which led to the products, and lyso-phospholipids formed by hydrolysis, which led to the low yields. The biocatalyst used was the commercial immobilized lipase Lipozyme TL IM and substrates used were phosphatidylcholine (PC) from soybean and caprylic acid. A response surface design was used to evaluate the influence of selected parameters and their relationships on the incorporation of caprylic acid and the corresponding recovery of PC. Incorporation of fatty acids increased with increasing enzyme dosage, reaction temperature, solvent amount, reaction time, and substrate ratio. Enzyme dosage had the most significant effect on the incorporation, followed by reaction time, reaction temperature, solvent amount, and substrate ratio. However the parameters had also a negative influence on the PC recovery. Solvent amount had the most negative effect on recovery, followed by enzyme dosage, temperature, and reaction time. Individually substrate ratio had no significant effect on the PC recovery. Interactions were observed between different parameters. On the basis of the models, the reaction was optimized for the maximum incorporation and maximum PC recovery. With all of the considerations, the optimal conditions are recommended as enzyme dosage 29%, reaction time 50 h, temperature 54 degrees C, substrate ratio 15 mol/mol caprylic acid/PC, and 5 mL of hexane per 3 g substrate. No additional water is necessary. Under these conditions, an incorporation of caprylic acid up to 46% and recovery of PC up to 60% can be obtained from the prediction. The prediction was confirmed from the verification experiments. PMID- 15801778 TI - Treatment of germinated wheat to increase levels of GABA and IP6 catalyzed by endogenous enzymes. AB - We found that the levels of bioactive products from wheat can be increased dramatically by manipulating germination conditions and taking advantage of the activity of endogenous enzymes. The yield of phytic acid (IP(6)) from wheat germinated in the presence of high, controlled levels of dissolved oxygen (188 +/ 28 mg/100 g wheat) was almost three times greater than that from wheat germinated with no supplemental oxygen (74 +/- 10 mg/100 g wheat). The yield of gamma-aminobutyric acid (GABA) from wheat germinated in the presence of uncontrolled levels of dissolved oxygen was 18 +/- 3 times greater than that from nonsupplemented wheat (1 mg/100 g wheat). The concentration of GABA was much greater in wheat germ than in whole wheat, and the yield of GABA from wheat germ processed with supplemental water (163 +/- 7 mg/100 g wheat germ) was notably greater than that from wheat germ processed with no supplemental water (100 +/- 2 mg/100 g wheat germ). In contrast, IP(6) was more concentrated in wheat bran, and the yield of IP(6) from wheat bran processed with supplemental water (3100 +/- 12 mg/100 g wheat bran) was notably higher than that from wheat bran processed with no supplemental water (2420 +/- 13 mg/100 g wheat bran). We conclude that the large amount of GABA extracted from wheat germ is likely due to high glutamate decarboxylase activity and low aminotransferase activity and that the large amount of IP(6) extracted from wheat bran is likely due to high levels of tyrosinase activity. Our findings indicate that bioactive molecules such as GABA and IP(6) can be successfully mass-produced by taking advantage of endogenous enzymatic activities. PMID- 15801779 TI - Asymmetrical synthesis of L-homophenylalanine using engineered Escherichia coli aspartate aminotransferase. AB - Site-directed mutagenesis was performed to change the substrate specificity of Escherichia coli aspartate aminotransferase (AAT). A double mutant, R292E/L18H, with a 12.9-fold increase in the specific activity toward L-lysine and 2-oxo-4 phenylbutanoic acid (OPBA) was identified. E. coli cells expressing this mutant enzyme could convert OPBA to L-homophenylalanine (L-HPA) with 97% yield and more than 99.9% ee using L-lysine as amino donor. The transamination product of L lysine, 2-keto-6-aminocaproate, was cyclized nonenzymatically to form Delta(1) piperideine 2-carboxylic acid in the reaction mixture. The low solubility of L HPA and spontaneous cyclization of 2-keto-6-aminocaproate drove the reaction completely toward L-HPA production. This is the first aminotransferase process using L-lysine as inexpensive amino donor for the L-HPA production to be reported. PMID- 15801780 TI - Hydrodynamic alterations during cyanobacteria (Arthrospira platensis) growth from low to high biomass concentration inside tubular photobioreactors. AB - The rheological behavior of an Arthrospira culture was studied from low to high biomass concentration. Two tubular undulating row photobioreactors (TURP-5r and TURP-10r), with a very short light path of 1.0 cm, were used during batch growth. In TURP-5r, the biomass concentration increased to 14.5 g(dw) L(-1), and alterations of the physical properties and hydrodynamic behavior occurred as a result. In the past, the rheological characteristics of photosynthetic-microbe cultures were rarely investigated because of the low biomass concentration attained in the systems. Developing closed photobioreactor technologies, the optimum biomass concentration rises and the viscosity, the generalized Reynolds number (N'(Re)), and the power required for culture recycling are also subject to alteration. Starting from a biomass concentration of 4.1 g(dw) L(-1), the Arthrospira culture already exhibits the characteristics of a non-Newtonian fluid. As a result of culture recycling from 2.0 to 20.5 g(dw) L(-1) and an available power of 1.67 W row(-1), we demonstrated that N'(Re) is reduced from 6265 to 1148. Our experimental results showed that N'(Re) of 2345 can be reached only at a cell concentration below 11.1 g(dw) L(-1), while at a cell concentration below 4.1 g(dw) L(-1) N'(Re) = 4080 was reached. The power consumption (P(c)) for culture recycling increased noticeably when the cell concentration rose; the highest P(c) increase attained was from 2.0 to 4.1 g(dw) L(-1). This is the range within which the Arthrospira culture changes from a Newtonian to a non-Newtonian fluid. PMID- 15801781 TI - A dual-growth kinetic model for biological wastewater reactors. AB - Biological wastewater reactors are traditionally divided into two groups based on modes of cell growth: suspension and attached (biofilm) growth. Kinetic descriptions of these reactors are based on confining cell growth to solid surfaces or void space. Because suspended cells grow in void space and biofilms grow on surfaces, both forms of microbial growth must in principle occur in a biological reactor, unless the surface is inhabitable by a biofilm. Cell growth and substrate utilization in both modes, suspension and attached, are fully accounted for in the model developed here. Simulations based on this model show that biofilms growing on the walls of a reactor, classified as a suspension culture, can contribute substantially to the total organics removal. Similarly, suspended cells in the voids of a "traditional biofilm" reactor can contribute significantly to degradation of organic substrates. The presence of biofilms can obviate total washout of suspended cells and avert reactor failure. Model simulations enable a comparison of attached and suspended biomass in terms of biomass accumulation, substrate degradation, and effectiveness of substrate utilization and illustrate interactions between the two forms of biomass. The model provides a unified way to analyze and design biological wastewater processes. PMID- 15801782 TI - Ca2+ is a cofactor required for membrane transport and maturation and is a yield determining factor in high cell density penicillin amidase production. AB - Penicillin amidases (PAs) from E. coli and A. faecalis are periplasmic enzymes that contain one tightly bound Ca(2+) per molecule that does not directly participate in the enzymatic function. This ion may, however, be required for the maturation of the pre-pro-enzyme. The pro-enzyme of homologous PAs are translocated through the Tat- (E. coli PA(EC)) and Sec- (A. faecalis PA(AF)) transport systems, respectively. Cell fractionation, electrophoresis, immunoblotting, and activity staining demonstrated that Ca(2+) binding is required for the membrane transport and maturation of the pro-enzyme to active enzyme. Pro-enzyme without Ca(2+) was targeted to the membrane but not translocated. Influence of Ca(2+) in medium and feed was studied for high cell density cultivations of E. coli expressing these enzymes. Without Ca(2+) in the feed the synthesis of the pre-pro-enzyme was hardly influenced. At optimal Ca(2+) content in the feed the active enzyme amount could be increased by 2 orders of magnitude up to 0.9 g/L (PA(EC)) and 2.3 g/L (PA(AF)) or 4% (PA(EC)) and 8% (PA(AF)) of the cell dry weight. The corresponding specific activities are 1700 U (PA(EC)) and 14000 U (PA(AF)) per gram cell dry weight, respectively. These values are higher than those published previously. Thus, for optimal yields of the studied and other extra- and periplasmic enzymes that require Ca(2+) or other ions as cofactors for membrane transport and maturation, sufficient cofactor must be added in the feed. PMID- 15801783 TI - Liquid water-ice I phase diagrams under high pressure: sodium chloride and sucrose models for food systems. AB - The knowledge of high pressure and low temperature phase diagrams of aqueous systems is required in fields such as food sciences, biology, cryo-microscopy and geology, to reduce processing costs, improve treatments results or advance in physical phenomena understanding. The phase transition curve between liquid water and ice I for sucrose and sodium chloride solutions has been obtained for concentrations ranging from 16% to 36% and from 1.63% to 16.09% (w/w), respectively. An accurate experimental method, based on the pressurization of an ice-solution mixture, adequate to build the entire phase transition curve at constant concentration, has been developed. Simon-like equations have been used to empirically describe the phase transition curves, so that they allow easy data interpolation. PMID- 15801784 TI - Inactivation of adenovirus type 5 by caustics. AB - Adenovirus shows significant promise as a vehicle for transfer of therapeutic genes into humans. Based on the importance of this viral vector, it is critical that adequate decontamination procedures are implemented during its large-scale production in multiproduct manufacturing facilities to prevent cross-product contamination and to reduce the risk of personnel exposure. Liquid decontamination procedures based on caustics are easily implemented in a manufacturing setting and are not corrosive to stainless steel surfaces at the concentrations found to inactivate viral proteins and nucleic acids. In this study, we have conducted small-scale experiments to determine the effectiveness of caustic inactivation procedures on adenovirus type 5 and have evaluated the robustness of the process to different sample matrices and adenovirus constructs. We find that the pH of a sample post-addition of caustic solution is a more accurate indicator of the effectiveness of the caustic than its concentration. We have demonstrated that a greater than 6 log reduction in the potency of adenovirus type 5 may be obtained upon exposure of the sample to sodium hydroxide and CIP-100 at concentrations greater than 0.09 M and 0.9%, respectively, at times greater than 10 min. PMID- 15801786 TI - Streamlining Escherichia coli S30 extract preparation for economical cell-free protein synthesis. AB - Escherichia coli extracts activate cell-free protein synthesis systems by providing the catalysts for translation and other supporting reactions. Recent results suggest that high-density fermentations can be used to provide the source cells, but the subsequent cell extract preparation procedure requires multiple centrifugation and dialysis steps as well as an expensive runoff reaction. In the work reported here, the extract preparation protocol duration was reduced by nearly 50% by significantly shortening several steps. In addition, by optimizing the runoff incubation, overall reagent costs were reduced by 70%. Nonetheless, extracts produced from the shorter, less expensive procedure were equally active. Crucial steps were further examined to indicate minimal ribosome loss during the standard 30,000g centrifugations. Furthermore, sucrose density centrifugation analysis indicated that although an incubation step significantly activates the extract, ribosome/polysome dissociation is not required. These insights suggest that consistent cell extract can be produced more quickly and with considerably less expense for large-scale cell-free protein production, especially when combined with high-density fermentation protocols. PMID- 15801785 TI - Selective isolation and purification of tat protein via affinity membrane separation. AB - This work deals with the separation of Tat protein from a complex fermentation broth using an affinity membrane system. Tat is a regulatory protein that is critical for HIV-1 replication and thus a potential candidate for vaccine and drug development. Furthermore, Tat can facilitate transport of exogenous molecules across cell membranes and is implicated in pathogenesis of HIV dementia. Affinity membranes were prepared through coupling of avidin within a 4 stack membrane construct. Tat (naturally biotinylated) accessibility in the bacterial lysate feed was influenced by the presence of RNAse, protein concentration, and ionic strength. Enhanced accessibility translated to a marked increase in the overall product yield per pass. The purity of the membrane isolated Tat was compared to that prepared via packed column chromatography through SDS-PAGE, Western blot, activity assay, and neurotoxicity studies. Tat protein produced via membrane separation yielded primarily monomeric forms of the oligopeptide sequence, whereas column chromatography produced predominately polymeric forms of Tat. These differences resulted in changes in the neurotoxicity and cellular uptake of the two preparations. PMID- 15801787 TI - Development of a purification process for adenovirus: controlling virus aggregation to improve the clearance of host cell DNA. AB - The clearance of host cell DNA is a critical goal for purification process development for recombinant Ad5 (rAd5) based vaccines and gene therapy products. We have evaluated the clearance of DNA by a rAd5 purification process utilizing nuclease digestion, ultrafiltration, and anion exchange (AEX) chromatography and found residual host cell DNA to consistently reach a limiting value of about 100 pg/10(11) rAd5 particles. Characterization of the purified rAd5 product using serial AEX chromatography, hydroxyapatite chromatography, or nuclease treatment with and without particle disruption showed that the residual DNA was associated with virus particles. Using a variety of additional physical characterization methods, a population of rAd5 virus in an aggregated state was detected. Aggregation was eliminated using nonionic detergents to attenuate hydrophobic interactions and sodium chloride to attenuate electrostatic interactions. After implementation of these modifications, the process was able to consistently reduce host cell DNA to levels at or below 5 pg/10(11) rAd5 particles, suggesting that molecular interactions between cellular DNA and rAd5 are important determinants of process DNA clearance capability and that the co-purifying DNA was not encapsidated. PMID- 15801788 TI - Antibody capture from corn endosperm extracts by packed bed and expanded bed adsorption. AB - Topical treatments of chronic infections with monoclonal antibodies will require large quantities of antibodies. Because plants have been proven capable of producing multisubunit antibodies and provide for large-scale production, they are likely hosts to enable such applications. Recovery costs must also be low because of the relatively high dosages required. Hence, we have examined the purification of a human secretory antibody from corn endosperm extracts by processing alternatives of packed bed and expanded bed adsorption (EBA). Because of the limited availability of the transgenic corn host, the system was modeled by adding the antibody to extracts of nontransgenic corn endosperm. Complete clarification of a crude extract followed by packed bed adsorption provided antibody product in 75% yield with 2.3-fold purification (with antibody accounting for 24% of total protein). The small size of the packed bed, cation exchange resin SP-Sepharose FF and the absence of a dense core (present in EBA resins) allowed for more favorable breakthrough performance compared to EBA resins evaluated. Four adsorbents specifically designed for EBA operation, with different physical properties (size and density), chemical properties (ligand), and base matrices were tested: SP-steel core resin (UpFront Chromatography), Streamline SP and Streamline DEAE (Amersham Biosciences), and CM Hyper-Z (BioSepra/Ciphergen Biosystems). Of these, the small hyperdiffuse-style resin from BioSepra had the most favorable adsorption characteristics. However, it could not be utilized with crude feeds due to severe interactions with corn endosperm solids that led to bed collapse. UpFront SP-steel core resin, because of its relatively smaller size and hence lower internal mass transfer resistance, was superior to the Streamline resins and operated successfully with application of a crude corn extract filtered to remove all solids of >44 microm. However, the EBA performance with this adsorbent provided a yield of only 61% and purification factor of 2.1 (with antibody being 22% of total protein). Process simulation showed that capital costs were roughly equal between packed and expanded bed processes, but the EBA design required four times greater operating expenditures. The use of corn endosperm as the starting tissue proved advantageous as the amount of contaminating protein was reduced approximately 80 times compared to corn germ and approximately 600 times compared to canola. Finally, three different inlet designs (mesh, glass beads, and mechanical mixing) were evaluated on the basis of their ability to produce efficient flow distribution as measured by residence time distribution analysis. All three provided adequate distribution (axial mixing was not as limiting as mass transfer to the adsorption process), while resins with different physical properties did not influence flow distribution efficiency values (i.e., Peclet number and HETP) when operated with the same inlet design. PMID- 15801789 TI - Decision-support tool for assessing biomanufacturing strategies under uncertainty: stainless steel versus disposable equipment for clinical trial material preparation. AB - This paper presents the application of a decision-support tool, SIMBIOPHARMA, for assessing different manufacturing strategies under uncertainty for the production of biopharmaceuticals. SIMBIOPHARMA captures both the technical and business aspects of biopharmaceutical manufacture within a single tool that permits manufacturing alternatives to be evaluated in terms of cost, time, yield, project throughput, resource utilization, and risk. Its use for risk analysis is demonstrated through a hypothetical case study that uses the Monte Carlo simulation technique to imitate the randomness inherent in manufacturing subject to technical and market uncertainties. The case study addresses whether start-up companies should invest in a stainless steel pilot plant or use disposable equipment for the production of early phase clinical trial material. The effects of fluctuating product demands and titers on the performance of a biopharmaceutical company manufacturing clinical trial material are analyzed. The analysis highlights the impact of different manufacturing options on the range in possible outcomes for the project throughput and cost of goods and the likelihood that these metrics exceed a critical threshold. The simulation studies highlight the benefits of incorporating uncertainties when evaluating manufacturing strategies. Methods of presenting and analyzing information generated by the simulations are suggested. These are used to help determine the ranking of alternatives under different scenarios. The example illustrates the benefits to companies of using such a tool to improve management of their R&D portfolios so as to control the cost of goods. PMID- 15801790 TI - Protein interactions in hydrophobic charge induction chromatography (HCIC). AB - A quantitative understanding of how proteins interact with hydrophobic charge induction chromatographic resins is provided. Selectivity on this mode of chromatography for monoclonal antibodies as compared to other model proteins is probed by means of a linear retention vs pH plot. The pH-dependent adsorption behavior on this mode of chromatography for a hydrophobic, charged solute is described by taking into account the equilibrium between a hydrophobic, charged solute and an ionizable, heterocyclic ligand. By analogy, an equation that is seen to adequately describe macromolecular retention under linear conditions over a range of pH is developed. A preparative, nonlinear isotherm that can capture both pH and salt concentration dependency for proteins is proposed by using an exponentially modified Langmuir isotherm model. This model is seen to successfully simulate adsorption isotherms for a variety of proteins over a range of pHs and mobile phase salt concentrations. Finally, the widely differing retention characteristics of two monoclonal antibodies are used to derive two different strategies for improving separations on this mode of chromatography. A better understanding of protein binding to this class of resins is seen as an important step to future exploitation of this mode of chromatography for industrial scale purification of proteins. PMID- 15801791 TI - Optimization of yield in magnetic cell separations using nickel nanowires of different lengths. AB - Ferromagnetic nanowires are shown to perform both high yield and high purity single-step cell separations on cultures of NIH-3T3 mouse fibroblast cells. The nanowires are made by electrochemical deposition in nanoporous templates, permitting detailed control of their chemical and physical properties. When added to fibroblast cell cultures, the nanowires are internalized by the cells via the integrin-mediated adhesion pathway. The effectiveness of magnetic cell separations using Ni nanowires 350 nm in diameter and 5-35 micrometers long in field gradients of 40 T/m was compared to commercially available superparamagnetic beads. The percent yield of the separated populations is found to be optimized when the length of the nanowire is matched to the diameter of the cells in the culture. Magnetic cell separations performed under these conditions achieve 80% purity and 85% yield, a 4-fold increase over the beads. This effect is shown to be robust when the diameter of the cell is changed within the same cell line using mitomycin-C. PMID- 15801792 TI - Influence of pH and ionic strength on the steric mass-action model parameters around the isoelectric point of protein. AB - The ion-exchange equilibrium and the dependence of the parameters in the steric mass-action (SMA) model on salt concentration and buffer pH around the isoelectric point of protein were studied. Bovine serum albumin (BSA, isoelectric point = 5.4) was used as a model protein and DEAE Sepharose FF as an ion exchanger. Finite batch adsorption experiments and isocratic elution chromatography were performed for the determination of the model parameters (i.e., characteristic charge, equilibrium constant, and steric factor). The results showed that pH had significant effects on the parameters. With an increase of pH from 4.5 to 6.5, the characteristic charge increased from 0.9 to 3.0 and leveled off as a plateau at pH above 5.5. The charge groups in the contact region of protein surface were considered to play a crucial role on the characteristic charge. The decrease of pH and increase of salt concentration lowered the absolute value of the zeta potential of the protein surface and led to a decrease of the equilibrium constant. The steric factor remained unchanged at about 31 at pH 5.5 and 6.0 and increased to 44.5 at pH 5.0 and 96.8 at pH 4.5, mainly as a result of the lower adsorption capacity of BSA at pH <5.5. Furthermore, the increase of the molecular volume of BSA at pH 4.5 would be an additional reason for the increase of the steric factor. Taking into account the effect of the pH and salt concentration on these parameters, the SMA model described the ion exchange equilibrium of protein more accurately. PMID- 15801793 TI - Production of soluble human interleukin-6 in cytoplasm by fed-batch culture of recombinant E. coli. AB - The major objective of this study is to identify fed-batch culture conditions optimal for the production of human interleukin-6 (hIL-6) in a soluble form. Five different expression vectors were constructed for the expression of hIL-6 and hIL 6s fused with NusA, maltose binding protein (MBP), thioredoxin (Trx) or ubiquitin (Ubi). A series of flask cultures were conducted in LB medium at 37 degrees C. The intact hIL-6 was expressed mostly in the form of inclusion body. More than 95% of the hIL-6 fused with NusA (NusA/hIL-6) and about 90% of MBP/hIL-6 were expressed in a soluble form, whereas Trx/hIL-6 and Ubi/hIL-6 were expressed mostly in the form of inclusion body. Based on this result, NusA was selected as the fusion partner for the production of hIL-6 in the subsequent experiments. A series of pH-stat fed-batch cultures of an E. coli BL21(DE3) transformed with a NusA/hIL-6 expression vector were conducted in a bioreactor with a working volume of about 3 L. As the amount of nitrogen source was increased in the feeding medium, more soluble NusA/hIL-6 was produced, while the total amount was not significantly changed. Under the best conditions tested, about 90% of NusA/hIL-6 was produced in the soluble form. In this case, the concentration of soluble NusA/hIL-6 was 7.5 g/L with a volumetric productivity of 0.43 g/L-h. PMID- 15801794 TI - Cellular fate of a modular DNA delivery system mediated by silica nanoparticles. AB - Development of efficient molecular medicines, including gene therapeutics, RNA therapeutics, and DNA vaccines, depends on efficient means of transfer of DNA or RNA into the cell. Potential problems, including toxicity and immunogenicity, surrounding viral methods of DNA delivery have necessitated the use of nonviral, synthetic carriers. To better design synthetic carriers, or transfection reagents, the modular design of viruses has inspired a modular approach to DNA and RNA delivery. Each modular component can be designed to circumvent each of the many barriers. The modular approach will allow modification of individual components for a specific application. By utilizing a dense silica nanoparticle to form a ternary complex, transfection efficiency of a DNA-transfection reagent complex was increased by a factor of approximately 10 by concentrating the DNA at the surface of cells. Surface modification of the silica nanoparticles allowed determination of the cellular uptake mechanism with only minor alteration of transfection efficiency. Nanoparticles are internalized by an endosome-lysosomal route followed by perinuclear accumulation. The modification mechanism confirms that surface modification of the modular system can allow specific moieties to be incorporated into the modular system without significant alteration of the transfection efficiency. By showing that the modular system based upon concentration of DNA at the level of the cell can be used to increase transfection efficiency, we have shown that further modification of the system may better target DNA delivery and overcome other barriers of DNA expression. PMID- 15801795 TI - Design and evaluation of a novel flow chamber for measuring cell adhesion to absorbable polymer films. AB - There is great interest in improving cellular attachment to synthetic materials, particularly for developing small diameter tissue-engineered vascular grafts. However, limited research has been conducted to evaluate the adhesion characteristics of different cell types to absorbable substrates. Tissue engineered vessels typically fail as a result of delamination of the endothelial cell layer when exposed to fluid or blood flow. The focus of this research was to design and evaluate a flow chamber, using fibroblasts, smooth muscle cells, and endothelial cells, to probe the bounds of the system. A flow chamber was designed and fabricated to compare the relative adhesion characteristics of cells to absorbable polymer films. A preliminary investigation of mouse fibroblast (3T3M) adhesion to semicrystalline poly-L-lactide (PLL) films was conducted to determine general operating specifications. Cell coverage on films was evaluated using a live-dead assay and image analysis; following exposure to flow, tests were similarly conducted. Based on these results, additional studies were conducted to compare the adhesion of rat aortic smooth muscle cells (SMC) and endothelial cells (EC) on PLL films. PMID- 15801796 TI - Methods to improve the yield and quality of DNA from dried and processed figs. AB - We describe here a molecular method that can be used to detect genome traits of a given horticultural item at each stage from the farm to the market. We developed a procedure to extract and amplify by PCR DNA obtained from complex matrixes, such as dried figs and fig jam. Few fragmented DNA molecules can be recovered from food products. However, we were able to increase the yield of PCR reactions by successfully applying an enzymatic repair protocol to retrieved DNA. PMID- 15801797 TI - Effect of copper sulfate on performance of a serum-free CHO cell culture process and the level of free thiol in the recombinant antibody expressed. AB - A recombinant Chinese hamster ovary (CHO) cell line was used to express a humanized antibody. Product quality analysis of this humanized antibody showed the presence of free thiol, due to unpaired cysteine residues in the Fab region. Decreased potency of this thiol Fab made it critical to minimize the levels of free thiol. In an effort to do this, we evaluated the effect of copper sulfate addition to the cell culture production medium. As a component of the production medium, copper sulfate can act as an oxidizing agent, thereby facilitating disulfide bond formation. Four concentrations of copper sulfate were added at the beginning of 2-L benchtop production cultures of the recombinant CHO cell line: 0, 5, 50, and 100 microM. We found that these copper sulfate additions had no effect on cell growth or antibody production. However, a slight dose-dependent depression in culture viability was observed. Analysis of the purified antibody showed that either the 50 or 100 microM copper sulfate additions reduced the level of free thiol by more than 10-fold. PMID- 15801798 TI - Evaluation of production parameters with the vaccinia virus expression system using microcarrier attached HeLa cells. AB - Parameters that affect production of the recombinant reporter protein, EGFP, in the T7 promoter based VOTE vaccinia virus-HeLa cell expression system were examined. Length of infection phase, inducer concentration, and timing of its addition relative to infection were evaluated in 6-well plate monolayer cultures. One hour infection with 1.0 mM IPTG added at the time of infection provided a robust process. For larger scale experiments, anchorage-dependent HeLa cells were grown on 5 g/L Cytodex 3 microcarriers. The change to this dynamic culture environment, with cell-covered microcarriers suspended in culture medium in spinner flasks, suggested a re-examination of the multiplicity of infection (MOI) for this culture type that indicated a need for an increase in the number of virus particles per cell to 5.0, higher than that needed for complete infection in monolayer tissue flask culture. Additionally, dissolved oxygen level and temperature during the protein production phase were evaluated for their effect on EGFP expression in microcarrier spinner flask culture. Both increased dissolved oxygen, based on surface area to volume (SA/V) adjustments, and decreased temperature from 37 to 31 degrees C showed increases in EGFP production over the course of the production phase. The level of production achieved with this system reached approximately 17 microg EGFP/10(6) infected cells. PMID- 15801799 TI - Effects of NHE1 expression level on CHO cell responses to environmental stress. AB - Ammonia, lactate and CO(2) inhibit animal cell growth. Accumulation of these metabolic byproducts also causes a decrease in intracellular pH (pH(i)). Transport systems regulate pH(i) in eukaryotic cells. Ion transporters have been cloned and overexpressed in cells but have not been examined for protection against the buildup of ammonia, lactate or CO(2). The Na(+)/H(+) exchangers (NHE) transport H(+) ions from cells during acidification to increase pH(i). We examined whether overexpression of NHE1 would provide CHO cells with greater protection from elevated ammonia, lactate or CO(2). NHE1 CHO cells were compared to MT2-1-8 ("normal" levels of NHE) and AP-1 (devoid of any NHE activity) CHO cell lines. Expression of at least "normal" levels of NHE1 is necessary for CHO cell survival during exposure to 30 mM lactic acid without pH adjustment or to 20 mM NH(4)Cl with pH adjustment. Resistance to an acute acid-load increased when NHE1 was overexpressed in CHO cells. Surprisingly, the inhibitory effect on cell growth at 195 mmHg pCO(2)/435 mOsm/kg (normal levels are 40 mmHg pCO(2)/ 320 mOsm/kg) was not affected by the NHE1 level. Also, there was no further decrease in CHO cell growth in the absence of NHE1 expression during elevated osmolality alone (up to 575 mOsm/kg). PMID- 15801800 TI - Spherical alginate granules formulated for quick-release active subtilisin. AB - Novel attrition-resistant and spherical enzyme granules encapsulating active subtilisin were formed by emulsification of 2% alginate sol loaded with active enzyme, instantaneous gelation triggered through in situ release of Ca(2+) (internal gelation), particle separation, and finally acetone extractive drying. Granular subtilisin was highly active, readily dispersible, and mechanically robust. This technique serves as a new and attractive alternative to established enzyme granulation processes, such as fluid bed coating, extrusion followed by marumerization, drum granulation, or prilling, for use in industrial enzyme applications such as detergents, textile manufacturing, and food processing. The formulation and encapsulation conditions were optimized to maximize the resistance of the granule to compression and impact forces, consistent with enzyme release and particle dispersion in detergent solutions. Well characterized alginates, with specified guluronic/mannuronic acid (G/M) content and molecular weight, were used in the formulation. The characteristics of the resulting microspheres, including their size and distribution, morphology, shrinkage, compression resistance, impact strength, solubility and encapsulation yield, were examined. Spherical dry granules were formulated with a mean diameter of 500 microm with particle sizes ranging from 300 to 800 microm. Dry alginate granules were discrete, spherical, and glossy white and exhibited impact strength, compression resistance, and solubility difference dependent on composition. Reduced starch levels, high alginate concentration, low alginate molecular weight, and use of high guluronate alginates resulted in the lowest dust level and highest compression resistance. Subtilisin mass yields were approximately 50%, and specific activity yields ranged from 60% to 100%. A formulation consisting of 3% SG150 alginate, 10% starch, 10% TiO(2), and 1% CaCO(3) provided granules appropriate for use in detergent application. PMID- 15801801 TI - Detection of point mutations in the HBV polymerase gene using a fluorescence intercalator in reverse micelles. AB - We report a novel and simple method for mutation detection in DNA oligonucleotides using a double-stranded DNA specific dye (SYBR Green I) in nanostructured molecular assemblies, called reverse micelles. The intercalation of SYBR Green I into the duplex DNA exhibits fluorescent emission in a CTAB/isooctane reverse micellar system as well as in an aqueous solution. We found marked differences in the fluorescence intensity between perfectly matched and mismatched 52-mer synthetic oligonucleotides, which were designed to contain the YMDD motif of the hepatitis B virus (HBV) polymerase gene, in a reverse micellar solution. Using this method, we successfully detected a mutation in PCR amplified oligonucleotides of the HBV polymerase gene in sera of four patients with chronic hepatitis B. This detection method does not require DNA immobilization, chemical modification of DNA, or any special apparatus; it only needs a normal fluorescence spectrophotometer, an inexpensive dye, and just 10 pmol of sample DNA. PMID- 15801802 TI - Plant cell culture monitoring using an in situ multiwavelength fluorescence probe. AB - A multiwavelength fluorescence probe is proposed for in situ monitoring of Eschscholtzia californica and Catharanthus roseus plant cell cultures. The potential of the probe as a tool for real-time estimation of biomass and production in secondary metabolites has been studied. The probe excitation range is 270-550 nm and the emission range is 310-590 nm, with a step of 20 nm for both excitation and emission filters. Many endogenous fluorophores such as NAD(P)H, riboflavins (riboflavin and derivatives such as FMN, FAD), tryptamine and tryptophan, and fluorescent secondary metabolites were analyzed simultaneously. NAD(P)H fluorescence signal (350/450 nm) showed to be an adequate signal for estimating cells activity. Riboflavins fluorescence signal (450/530 nm) followed C. roseus cell concentration both for the growth phase and after elicitation with jasmonic acid. Fluorescence from the alkaloids interfered with NAD(P)H signal during the production phase. For C. roseus, tryptophan, tryptamine, ajmalicine and serpentine were monitored by the probe. For E. californica, fluorescence from alkaloids overlapped with riboflavins preventing from using the probe to follow cell growth but global alkaloids production could be followed using the probe. PMID- 15801803 TI - A new integrated membrane filtration and chromatographic device. AB - To improve protein separation, a novel integrated device combining membrane filtration and chromatography has been developed. The device basically consists of a hollow fiber filtration module whose shell side is filled with chromatographic resin beads. However, there is an essentially impermeable coated zone near the hollow fiber module outlet. The integrated device enjoys the advantages of both membrane filtration and chromatography; it also allows one to load the chromatographic media directly from the fermentation broth or lysate and separate the adsorbed proteins through the subsequent elution step in a cyclic process. Interfacial polymerization was carried out to coat the bottom section of the hollow fiber membrane; the rest of the hollow fiber membrane remained unaffected. Myoglobin (Mb) and alpha-lactalbumin (alpha-LA) were primarily used as model proteins in a binary mixture; binary mixtures of Mb and bovine serum albumin (BSA) were also investigated. Separation behaviors of binary protein mixtures were studied in devices having either an ultrafiltration (UF) or a microfiltration (MF) membrane. Experimental results show that the breakthrough time and the protein loading capacities were dramatically improved after introducing the impermeable coating in both UF and MF modules. For a synthetic yeast fermentation broth feed, four loading-washing-elution-reequilibration-based cyclic runs for separation of Mb and alpha-LA were performed in the device using a MF membrane with a coated zone without cleaning in between. The Mb and alpha-LA elution profiles for the four consecutive runs were almost superimposable. Due to lower transmembrane flux in this device plus the periodical washing-elution during the chromatographic separation, fouling was not a problem, unlike in conventional microfiltration. PMID- 15801804 TI - Production of glucose syrups in highly concentrated systems. AB - We have investigated the hydrolysis of maltodextrins in a high concentration (up to 70%), by means of enzymatic and acid catalysis. The study revealed that the equilibrium compositions of the catalyzed reactions were kinetically determined by the selectivity of the catalyst, the substrate concentration and the reaction time. A model comprising a set of two kinetic equations was used to describe the hydrolysis and condensation reactions of glucoamylase-catalyzed reactions, even to highly concentrated systems. Increased substrate concentration resulted in the formation of more condensation products. The enzyme inhibition was low and was found to be independent of the substrate concentration. PMID- 15801805 TI - Combination of extractive solvent addition and immobilization culture for continuous production of scopoletin by tobacco cells. AB - Extractive solvent addition was combined with immobilization cultures of Nicotiana tabacum cells to produce scopoletin. Using various solvents, the partition coefficients of scopoletin between the solvent and water phases and the solvent toxicity to the cell viability were investigated. The effect of the solvent addition on cell growth and scopoletin production was elucidated in the suspension cultures. Coconut oil, one of the natural vegetable oils, was selected as the most suitable extractive solvent. The cells were immobilized in the calcium alginate gel bead coated with a cell-free gel film and then the batch cultures with the addition of various volumes of the coconut oil were performed. The total scopoletin production increased with the solvent volume according to the amount of scopoletin transferred from the medium to the solvent. The maximum productivity obtained in the batch immobilization cultures was about 16 times larger than that in the suspension culture without solvent. A continuous production system, in which the fresh solvent was supplied to the culture system and the solvent containing scopoletin was recovered from it, was constructed. The integrated scopoletin production in the effluent oil attained 2.21 mg/gDCW for 30 days at 100 cm(3)/day without cell leakage. PMID- 15801806 TI - Enhanced cell-free protein synthesis using a S30 extract from Escherichia coli grown rapidly at 42 degrees C in an amino acid enriched medium. AB - Growths of Escherichia coli strain A19 were investigated in a 5-L fermentor at 37 and 42 degrees C either in Pratt's medium (a standard medium for cell-free protein synthesis using its S30 extract) or in a casamino acids supplemented Pratt's medium (aa-enriched medium). Specific growth rates in Pratt's medium at 37 and 42 degrees C were 0.77 and 0.46 h(-1), respectively, whereas those in the aa-enriched medium at 37 and 42 degrees C were 0.87 and 1.49 h(-1), respectively. The extent of cell-free chloramphenicol acetyltransferase (CAT) synthesis was compared at 37 degrees C incubation (from a plasmid pK7-CAT) for S30 extracts prepared from the cells cultured in the aa-enriched medium at 37 or 42 degrees C. A 40% increase in CAT synthesis occurred when the 42 degrees C/S30 extract was used as compared with 37 degrees C/S30 extract. CAT and both the light and heavy chains (Lc and Hc) of the Fab fragment of an antibody 6D9 were synthesized at 37 degrees C in the cell-free synthesis in the presence of [(14)C]Leu. Their reaction mixtures were subjected to SDS-PAGE autoradiographic analysis. It was found that most of the synthesized proteins were in the soluble fraction when 42 degrees C/S30 extract was used, suggesting that the 42 degrees C/S30 extract contained greater amounts of various protein folding factors. A dialysis membrane minibioreactor with a reaction volume ca. 0.5 mL was handmade by the authors. The advantages of the minibioreactor are a simple configuration, a low manufacturing cost, and the capability of the dialysis membrane replacement. Increased CAT synthesis was also observed for continuous exchange cell-free (CECF) protein synthesis at 37 degrees C when the 42 degrees C/S30 extract was used in the minibioreactor. Some plausible reasons to give higher protein synthesis activity of the 42 degrees C/S30 extract are discussed. PMID- 15801807 TI - Evidence for yeast autophagy during simulation of sparkling wine aging: a reappraisal of the mechanism of yeast autolysis in wine. AB - Yeast autolysis is the source of several molecules responsible for the quality of wines aged in contact with yeast cells. However, the mechanisms of yeast autolysis during wine aging are not completely understood. All descriptions of yeast autolysis in enological conditions emphasize the disturbance of cell organization as the starting event in the internal digestion of the cell, while no reference to autophagy is found in wine-related literature. By using yeast mutants defective in the autophagic or the Cvt pathways we have demonstrated that autophagy does take place in wine production conditions. This finding has implications for the genetic improvement of yeasts for accelerated autolysis. PMID- 15801808 TI - Heterologous protein production from the inducible MET25 promoter in Saccharomyces cerevisiae. AB - Heterologous protein production late in Saccharomyces cerevisiae fermentations is often desirable because it may help avoid the unintentional selection of more rapidly growing, non-protein-expressing cells or allow for the expression of toxic proteins. Here, we describe the use of the MET25 promoter for the production of human serum albumin (HSA) and HSA-fusion proteins in S. cerevisiae. In media lacking methionine, the MET25 promoter yielded high expression levels of HSA and HSA fused to human glucagon, human growth hormone, human interferon alpha, and human interleukin-2. More importantly, we have shown that this system can be used to delay heterologous protein production until late log phase of the growth of the culture and does not require the addition of an exogenous inducer. PMID- 15801809 TI - A generalized model for settling velocity of aerobic granular sludge. AB - Aerobic granulation is a novel biotechnology recently receiving intensive research attention. Aerobic granules developed in SBR can be as big as several millimeters, thus the traditional models describing the settling velocity of activated sludge are no long valid in aerobic granules culture. In this study, a new type of model was developed for the settling velocity of aerobic granules. This model shows that the settling velocity of aerobic granules is the function of SVI, mean size of granules and biomass concentration of granules. When the size of bioparticle is small enough, the proposed model reduces to the well-known Vesilind equation. Results indicated that the proposed model could satisfactorily fit experimental results obtained in the course of aerobic granulation under different conditions, while the Vesilind equation failed to or very poorly fit the experimental data. In addition, the proposed model can also be extended to anaerobic granules. The settling velocity is one of the most important parameters in both aerobic and anaerobic granulation, and successful biogranulation is highly related to the manipulation of settling velocity. It was demonstrated that the proposed model can sever as a useful tool for design and operation engineers to properly select the settling velocity for enhanced aerobic and anaerobic granulation. PMID- 15801810 TI - Redistribution of metabolic fluxes in the central aerobic metabolic pathway of E. coli mutant strains with deletion of the ackA-pta and poxB pathways for the synthesis of isoamyl acetate. AB - Although the bacterium E. coli is chosen as the host in many bioprocesses, products derived from the central aerobic metabolic pathway often compete with the acetate-producing pathways poxB and ackA-pta for glucose as the substrate. As such, a significant portion of the glucose may be excreted as acetate, wasting substrate that could have otherwise been used for the desired product. The production of the ester isoamyl acetate from acetyl-CoA by ATF2, a yeast alcohol acetyl transferase, was used as a model system to demonstrate the beneficial effects of reducing acetate production. All strains tested for ester production also overexpressed panK, a native E. coli gene that previous studies have shown to increase free intracellular CoA levels when fed with pantothenic acid. A recombinant E. coli strain with a deletion in ackA-pta produces less acetate and more isoamyl acetate than the wild-type E. coli strain. When both acetate producing pathways were deleted, the acetate production was greatly reduced. However, pyruvate began to accumulate, so that the overall ester production remained largely unchanged. To produce more ester, a previously established strategy of increasing the flux from pyruvate to acetyl-CoA was adopted by overexpressing pyruvate dehydrogenase. The ester production was then 80% higher in the poxB, ackA-pta strain (0.18 mM) than that found in the single ackA-pta mutant (0.10 mM), which also overexpressed PDH. PMID- 15801811 TI - Production of nonclassical inclusion bodies from which correctly folded protein can be extracted. AB - Human granulocyte-colony stimulating factor (hG-CSF), an important biopharmaceutical drug used in oncology, is currently produced mainly in Escherichia coli. Expression of human hG-CSF gene in E. coli is very low, and therefore a semisynthetic, codon-optimized hG-CSF gene was designed and subcloned into pET expression plasmids. This led to a yield of over 50% of the total cellular proteins. We designed a new approach to biosynthesis at low temperature, enabling the formation of "nonclassical" inclusion bodies from which correctly folded protein can be readily extracted by nondenaturing solvents, such as mild detergents or low concentrations of polar solvents such as DMSO and nondetergent sulfobetaines. FT-IR analysis confirmed different nature of inclusion bodies with respect to the growth temperature and indicated presence of high amounts of very likely correctly folded reduced hG-CSF in nonclassical inclusion bodies. The yield of correctly folded, functional hG-CSF obtained in this way exceeded 40% of the total hG-CSF produced in the cells and is almost completely extractable under nondenaturing conditions. The absence of the need to include a denaturation/renaturation step in the purification process allows the development of more efficient processes characterized by higher yields and lower costs and involving environment-friendly technologies. The technology presented works successfully at the 50-L scale, producing nonclassical inclusion bodies of the same quality. The approach developed for the production of hG-CSF could be extended to other proteins; thus, a broader potential for industrial exploitation is envisaged. PMID- 15801812 TI - Amino Acid esters prevent thermal inactivation and aggregation of lysozyme. AB - Small potent inhibitors of aggregation are eagerly demanded for preventing the inactivation of proteins. This paper shows that amino acid esters (AAEs) prevent heat-induced aggregation and inactivation of hen egg lysozyme. Lysozyme was completely inactivated (<1% original activity) during heat treatment at 98 degrees C for 30 min in a solution containing 0.2 mg/mL lysozyme in 50 mM Na phosphate buffer (pH 6.5). The residual activities only slightly increased (<5%) in the presence of 100 mM commonly used additives such as arginine, guanidine, urea, and sugars. However, in the presence of 100 mM AAEs, the residual activities were >60% and no aggregates were observed during the heat treatment at 98 degrees C for 30 min. This fact provides new information on the scaffold for designing additives to prevent heat-induced aggregation. PMID- 15801813 TI - Cell chromatography: separation of different microbial cells using IMAC supermacroporous monolithic columns. AB - Supermacroporous monolithic columns with Cu(2+)-IDA ligands have been successfully used for chromatographic separation of different types of microbial cells. The bed of monolithic matrix is formed by a cryogel of poly(acrylamide) cross-linked with methylenebis(acrylamide) and has a network of large (10-100 microm) interconnected pores allowing unhindered passage of whole cells through the plain cryogel column containing no ligands. Two model systems have been studied: the mixtures of wild-type Escherichia coli (w.t. E. coli) and recombinant E. coli cells displaying poly-His peptides (His-tagged E. coli) and of w.t. E. coli and Bacillus halodurans cells. Wild-type E. coli and His-tagged E. coli were quantitatively captured from the feedstock containing equal amounts of both cell types and recovered by selective elution with imidazole and EDTA, with yields of 80% and 77%, respectively. The peak obtained after EDTA elution was 8-fold enriched with His-tagged E. coli cells as compared with the peak from imidazole elution, which contained mainly weakly bound w.t. E. coli cells. Haloalkalophilic B. halodurans cells had low affinity to the Cu(2+)-IDA cryogel column and could be efficiently separated from a mixture with w.t. E. coli cells, which were retained and recovered in high yields from the column with imidazole gradient. All the cells maintained their viability after the chromatographic procedure. The results show that chromatography on affinity supermacroporous monolithic columns is a promising approach to efficient separations of individual cell types. PMID- 15801814 TI - Advance and prospect of bionanomaterials. PMID- 15801815 TI - Adverse cardiovascular effects of the coxibs. PMID- 15801816 TI - Discovery of a fluoroindolo[2,3-a]carbazole clinical candidate with broad spectrum antitumor activity in preclinical tumor models superior to the marketed oncology drug, CPT-11. AB - A series of fluoroglycosylated fluoroindolocarbazoles was examined with respect to their topoisomerase I activity, cytotoxicity, and selectivity. The lead clinical candidate from this series, BMS-250749, displays broad spectrum antitumor activity superior to CPT-11 against some preclinical xenograft models, including curative antitumor activity against Lewis lung carcinoma. PMID- 15801817 TI - Discovery of a novel series of peroxisome proliferator-activated receptor alpha/gamma dual agonists for the treatment of type 2 diabetes and dyslipidemia. AB - A series of 2-aryloxy-2-methyl-propionic acid compounds and related analogues were designed, synthesized, and evaluated for their PPAR agonist activities. 2 [(5,7-Dipropyl-3-trifluoromethyl)-benzisoxazol-6-yloxy]-2-methylpropionic acid (4) was identified as a PPARalpha/gamma dual agonist with relative PPARalpha selectivity and demonstrated potent efficacy in lowering both glucose and lipids in animal models without causing body weight gain. The PPARalpha activity of 4 appeared to have played a significant role in lowering glucose levels in db/db mice. PMID- 15801818 TI - 3-Substituted imidazo[1,2-d][1,2,4]-thiadiazoles: a novel class of factor XIIIa inhibitors. AB - A new class of selective FXIIIa inhibitors with a bicyclic [1,2,4]-thiadiazole pharmacophore is described. At 160 muM, compound 8 caused 50% reduction in fibrin gamma-chain cross-linking and suppressed the polymerization of alpha chains in platelet-depleted human plasma clots. Fibrinolysis rates in response to tissue plasminogen activator were directly proportional to the concentration of 8 in plasma at the time of clotting. PMID- 15801819 TI - Design of potent and selective 2-aminobenzimidazole-based p38alpha MAP kinase inhibitors with excellent in vivo efficacy. AB - We report the design and discovery of a 2-aminobenzimidazole-based series of potent and highly selective p38alphainhibitors. The lead compound 1 had low nanomolar activity in both ATP competitive enzyme binding and inhibition of TNFalpha release in macrophages. Compound 18 showed excellent pharmacokinetics properties and oral activity in the rat collagen induced arthritis model compared with other p38 reference compounds. A SAR strategy to address CyP3A4 liability is also described. PMID- 15801820 TI - Discovery of bicycloalkyl urea melanin concentrating hormone receptor antagonists: orally efficacious antiobesity therapeutics. AB - Melanin concentrating hormone (MCH) is involved in regulation of food intake and energy homeostasis. Antagonists of the MCH receptor are expected to affect food intake and weight gain, making MCH-R1 an attractive target for obesity treatment. Herein, we report the discovery of a novel, orally active series of MCH-R1 antagonists that exhibit in vivo efficacy in rodent obesity models. PMID- 15801821 TI - Virtual docking approaches to protein kinase B inhibition. AB - We examined some in silico approaches to identify Akt (protein kinase B) inhibitors. Experimental validation of selected compounds was achieved using a fluorescence-based enzymatic assay and a substrate phosphorylation assay involving the protein GSK-3. We report on success and failure obtained by using several strategies including FlexX, GOLD, and CSCORE, where the 100-200 top scoring compounds from a 50000-compound library were experimentally tested. This study led to the identification of low micromolar Akt1 inhibitors. PMID- 15801822 TI - 9-hydroxyazafluorenes and their use in thrombin inhibitors. AB - Optimization of a previously reported thrombin inhibitor, 9-hydroxy-9 fluorenylcarbonyl-l-prolyl-trans-4-aminocyclohexylmethylamide (1), by replacing the aminocyclohexyl P1 group provided a new lead structure, 9-hydroxy-9 fluorenylcarbonyl-l-prolyl-2-aminomethyl-5-chlorobenzylamide (2), with improved potency (K(i) = 0.49 nM for human thrombin, 2x APTT = 0.37 microM in human plasma) and pharmacokinetic properties (F = 39%, iv T(1/2) = 13 h in dogs). An effective strategy for reducing plasma protein binding of 2 and improving efficacy in an in vivo thrombosis model in rats was to replace the lipophilic fluorenyl group in P3 with an azafluorenyl group. Systematic investigation of all possible azafluorenyl P3 isomers and azafluorenyl-N-oxide analogues of 2 led to the identification of an optimal compound, 3-aza-9-hydroxyfluoren-9(R)-ylcarbonyl l-prolyl-2-aminomethyl-5-chlorobenzylamide (19b), with high potency (K(i) = 0.40 nM, 2x APTT = 0.18 microM), excellent pharmacokinetic properties (F = 55%, T(1/2) = 14 h in dogs), and complete efficacy in the in vivo thrombosis model in rats (inhibition of FeCl(3)-induced vessel occlusions in six of six rats receiving an intravenous infusion of 10 microg/kg/min of 19b). The stereochemistry of the azafluorenyl group in 19b was determined by X-ray crystallographic analysis of its N-oxide derivative (23b) bound in the active site of human thrombin. PMID- 15801824 TI - Binding mode prediction of cytochrome p450 and thymidine kinase protein-ligand complexes by consideration of water and rescoring in automated docking. AB - The popular docking programs AutoDock, FlexX, and GOLD were used to predict binding modes of ligands in crystallographic complexes including X-ray water molecules or computationally predicted water molecules. Isoenzymes of two different enzyme systems were used, namely cytochromes P450 (n = 19) and thymidine kinases (n = 19) and three different "water" scenarios: i.e., docking (i) into water-free active sites, (ii) into active sites containing crystallographic water molecules, and (iii) into active sites containing water molecules predicted by a novel approach based on the program GRID. Docking accuracies were determined in terms of the root-mean-square deviation (RMSD) accuracy and, newly defined, in terms of the ligand catalytic site prediction (CSP) accuracy. Consideration of both X-ray and predicted water molecules and the subsequent pooling and rescoring of all solutions (generated by all three docking programs) with the SCORE scoring function significantly improved the quality of prediction of the binding modes both in terms of RMSD and CSP accuracy. PMID- 15801823 TI - Structure-activity relationships of pregabalin and analogues that target the alpha(2)-delta protein. AB - Pregabalin exhibits robust activity in preclinical assays indicative of potential antiepileptic, anxiolytic, and antihyperalgesic clinical efficacy. It binds with high affinity to the alpha(2)-delta subunit of voltage-gated calcium channels and is a substrate of the system L neutral amino acid transporter. A series of pregabalin analogues were prepared and evaluated for their alpha(2)-delta binding affinity as demonstrated by their ability to inhibit binding of [(3)H]gabapentin to pig brain membranes and for their potency to inhibit the uptake of [(3)H]leucine into CHO cells, a measure of their ability to compete with the endogenous substrate at the system L transporter. Compounds were also assessed in vivo for their ability to promote anxiolytic, analgesic, and anticonvulsant actions. These studies suggest that distinct structure activity relationships exist for alpha(2)-delta binding and system L transport inhibition. However, both interactions appear to play an important role in the in vivo profile of these compounds. PMID- 15801825 TI - Enantiospecificity of glutamate carboxypeptidase II inhibition. AB - Two representative glutamate carboxypeptidase II (GCP II) inhibitors, 2 (hydroxypentafluorophenylmethyl-phosphinoylmethyl)pentanedioic acid 2 and 2-(3 mercaptopropyl)pentanedioic acid 3, were synthesized in high optical purities (>97%ee). The two enantiomers of 2 were prepared from previously reported chiral intermediates, (R)- and (S)-2-(hydroxyphosphinoylmethyl)pentanedioic acid benzyl esters 8. The synthesis of (R)- and (S)-3 involves the hydrolysis of (R)- and (S) 3-(2-oxo-tetrahydro-thiopyran-3-yl)propionic acids, (R)- and (S)-11, the corresponding optically pure thiolactones delivered by chiral chromatographic separation of the racemic 11. GCP II inhibitory assay revealed that (S)-2 is 40 fold more potent than (R)-2. In contrast, both enantiomers of 3 inhibited GCP II with nearly equal potency. The efficacy observed in subsequent animal studies with these enantiomers correlated well with the inhibitory potency in a GCP II assay. PMID- 15801826 TI - A knowledge-based energy function for protein-ligand, protein-protein, and protein-DNA complexes. AB - We developed a knowledge-based statistical energy function for protein-ligand, protein-protein, and protein-DNA complexes by using 19 atom types and a distance scale finite ideal-gas reference (DFIRE) state. The correlation coefficients between experimentally measured protein-ligand binding affinities and those predicted by the DFIRE energy function are around 0.63 for one training set and two testing sets. The energy function also makes highly accurate predictions of binding affinities of protein-protein and protein-DNA complexes. Correlation coefficients between theoretical and experimental results are 0.73 for 82 protein protein (peptide) complexes and 0.83 for 45 protein-DNA complexes, despite the fact that the structures of protein-protein (peptide) and protein-DNA complexes were not used in training the energy function. The results of the DFIRE energy function on protein-ligand complexes are compared to the published results of 12 other scoring functions generated from either physical-based, knowledge-based, or empirical methods. They include AutoDock, X-Score, DrugScore, four scoring functions in Cerius 2 (LigScore, PLP, PMF, and LUDI), four scoring functions in SYBYL (F-Score, G-Score, D-Score, and ChemScore), and BLEEP. While the DFIRE energy function is only moderately successful in ranking native or near native conformations, it yields the strongest correlation between theoretical and experimental binding affinities of the testing sets and between rmsd values and energy scores of docking decoys in a benchmark of 100 protein-ligand complexes. The parameters and the program of the all-atom DFIRE energy function are freely available for academic users at http://theory.med.buffalo.edu. PMID- 15801827 TI - Structures of three classes of anticancer agents bound to the human topoisomerase I-DNA covalent complex. AB - Human topoisomerase I (top1) is the molecular target of a diverse set of anticancer compounds, including the camptothecins, indolocarbazoles, and indenoisoquinolines. These compounds bind to a transient top1-DNA covalent complex and inhibit the resealing of a single-strand nick that the enzyme creates to relieve superhelical tension in duplex DNA. (Hertzberg, R. P.; et al. Biochem. 1989, 28, 4629-4638. Hsiang, Y. H.; et al. J. Biol. Chem 1985, 260, 14873-14878. Champoux, J. J. Annu. Rev. Biochem. 2001, 70, 369-413. Stewart, L.; et al. Science 1998, 729, 1534-1541.) We report the X-ray crystal structures of the human top1-DNA complex bound with camptothecin and representative members of the indenoisoquinoline and indolocarbazole classes of top1 poisons. The planar nature of all three structurally diverse classes allows them to intercalate between DNA base pairs at the site of single-strand cleavage. All three classes of compounds have a free electron pair near Arg364, a residue that if mutated confers resistance to all three classes of drugs. The common intercalative binding mode is augmented by unexpected chemotype-specific contacts with amino acid residues Asn352 and Glu356, which adopt alternative side-chain conformations to accommodate the bound compounds. These new X-ray structures explain how very different molecules can stabilize top1-DNA covalent complexes and will aid the rational design of completely novel structural classes of anticancer drugs. PMID- 15801828 TI - Novel cyano- and N-isopropylamidino-substituted derivatives of benzo[b]thiophene 2-carboxanilides and benzo[b]thieno[2,3-c]quinolones: synthesis, photochemical synthesis, crystal structure determination, and antitumor evaluation. 2. AB - Derivatives of 3-chlorobenzo[b]thiophene-2-carboxanilides and their "cyclic" analogues benzo[b]thieno[2,3-c]quinolones were synthesized. Spectroscopic study of the interactions of some representatives of "cyclic" derivatives and their "acyclic" precursors with ds-DNA/RNA supported strong intercalative binding of the former and weak nonintercalative binding of the latter group of compounds. All tested compounds showed a certain antiproliferative effect on a series of human tumor cells and on a normal cell line. Among the compounds, those with one amidino-substituent have shown the best effect. The most active benzo[b]thieno[2,3-c]quinolones induced apparent S and G2/M arrests of the cell cycle, which resulted in apoptosis. These results strongly suggest that the compounds may act as topoisimerase "poisons", which is in good agreement with their intercalative mode of binding to ds-DNA/RNA, in contrast to the studied "acyclic"group of derivatives. 6a and 6d showed the best selectivity by inhibiting the growth of tumor cells but not of normal fibroblasts. PMID- 15801829 TI - A comparison of the binding sites of matrix metalloproteinases and tumor necrosis factor-alpha converting enzyme: implications for selectivity. AB - MMPs and TACE (ADAM-17) assume independent, parallel, or opposite pathological roles in cancer, arthritis, and several other diseases. For therapeutic purposes, selective inhibition of individual MMPs and TACE is required in most cases due to distinct roles in diseases and the need to preserve activities in normal states. Toward this goal, we compared force-field interaction energies of five ubiquitous inhibitor atoms with flexible binding sites of 24 known human MMPs and TACE. The results indicate that MMPs 1-3, 10, 11, 13, 16, and 17 have at least one subsite very similar to TACE. S3 subsite is the best target for development of specific TACE inhibitors. Specific binding to TACE compared to most MMPs is promoted by placing a negatively charged ligand part at the bottom of S2 subsite, at the entrance of S1' subsite, or the part of S3' subsite that is close to catalytic zinc. Numerous other clues, consistent with available experimental data, are provided for design of selective inhibitors. PMID- 15801830 TI - Pyrido[2,3-d]pyrimidin-7-ones as specific inhibitors of cyclin-dependent kinase 4. AB - Inhibition of the cell cycle kinase, cyclin-dependent kinase-4 (Cdk4), is expected to provide an effective method for the treatment of proliferative diseases such as cancer. The pyrido[2,3-d]pyrimidin-7-one template has been identified previously as a privileged structure for the inhibition of ATP dependent kinases, and good potency against Cdks has been reported for representative examples. Obtaining selectivity for individual Cdk enzymes, particularly Cdk4, has been challenging. Here, we report that the introduction of a methyl substituent at the C-5 position of the pyrido[2,3-d]pyrimidin-7-one template is sufficient to confer excellent selectivity for Cdk4 vs other Cdks and representative tyrosine kinases. Further optimization led to the identification of highly potent and selective inhibitors of Cdk4 that exhibit potent antiproliferative activity against human tumor cells in vitro. The most selective Cdk4 inhibitors were evaluated for antitumor activity against MDA-MB-435 human breast carcinoma xenografts in mice. PMID- 15801831 TI - Discovery of a potent and selective inhibitor of cyclin-dependent kinase 4/6. AB - A pharmacological approach to inhibition of cyclin-dependent kinases 4 and 6 (Cdk4/6) using highly selective small molecule inhibitors has the potential to provide novel cancer therapies for clinical use. Achieving high levels of selectivity for Cdk4/6, versus other ATP-dependent kinases, presents a significant challenge. The pyrido[2,3-d]pyrimidin-7-one template provides an effective platform for the inhibition of a broad cross-section of kinases, including Cdks. It is now demonstrated that the modification of pyrido[2,3 d]pyrimidin-7-ones to include a 2-aminopyridine side chain at the C2-position provides inhibitors with exquisite selectivity for Cdk4/6 in vitro. This selectivity profile is recapitulated in cells where the most selective inhibitors create a G(1) block at concentrations up to 100-fold the IC(50) for cell proliferation. On the basis of its selectivity profile and pharmacokinetic profile, compound 43 (PD 0332991) was identified as a drug candidate for the treatment of cancer. PMID- 15801832 TI - Sulfur-substituted alpha-alkyl phenethylamines as selective and reversible MAO-A inhibitors: biological activities, CoMFA analysis, and active site modeling. AB - A series of phenethylamine derivatives with various ring substituents and with or without N-methyl and/or C-alpha methyl or ethyl groups was synthesized and assayed for their ability reversibly to inhibit monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B). Several compounds showed potent and selective MAO-A inhibitory activity (IC(50) in the submicromolar range) but none showed appreciable activity toward MAO-B. A three-dimensional quantitative structure activity relationship study for MAO-A inhibition was performed on the series using comparative molecular field analysis (CoMFA). The resulting model gave a cross-validated q(2) of 0.72 and showed that in this series of compounds steric properties of the substituents were more important than electrostatic effects. Molecular modeling based on the recently published crystal structure of inhibitor bound MAO-A provided detailed evidence for specific interactions of the ligands with the enzyme, supported by previous references and consistent with results from the CoMFA. On the basis of these results, structural determinants for selectivity of substituted amphetamines for MAO-A are discussed. PMID- 15801833 TI - Synthesis of 3-arylpiperazinylalkylpyrrolo[3,2-d]pyrimidine-2,4-dione derivatives as novel, potent, and selective alpha1-adrenoceptor ligands. AB - Novel compounds characterized by a pyrrolo[3,2-d]pyrimidine-2,4-dione (PPm) system connected through an alkyl chain to a phenylpiperazine (PPz) residue were designed as structural analogues of the alpha(1)-adrenoceptor (alpha(1)-AR) ligand RN5 (1). In this new series of derivatives an arylpyrrolo moiety has replaced the indole nucleus of RN5. Several structural modifications were performed on the PPm and PPz moieties and the connecting alkyl chain. These compounds were synthesized and tested in radioligand binding experiments where many of them showed interesting binding profiles. Some compounds, including 31, 34, and 36, displayed substantial alpha(1)-AR selectivity with respect to serotoninergic 5-HT(1A) and dopaminergic D(1) and D(2) receptors. Two different molecular modeling approaches (pharmacophoric mapping and quantitative structure affinity relationship analysis) have been applied to rationalize, at a quantitative level, the relationships between affinity toward alpha(1)-ARs and the structure of the studied compounds. Several QSAR models have been reported and described, accounting for the influence of various molecular portions on such affinity data. PMID- 15801834 TI - Hierarchical database screenings for HIV-1 reverse transcriptase using a pharmacophore model, rigid docking, solvation docking, and MM-PB/SA. AB - In this work, an efficient strategy was presented to search drug leads for human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) using hierarchical database screenings, which included a pharmacophore model, multiple-conformation rigid docking, solvation docking, and molecular mechanics-Poisson Boltzmann/surface area (MM-PB/SA) sequentially. Encouraging results were achieved in searching a refined available chemical directory (ACD) database: the enrichment factor after the first three filters was estimated to be 25-fold; the hit rate for all the four filters was predicted to be 41% in a control test using 37 known HIV-1 non-nucleoside reverse transcriptase inhibitors; 10 out of 30 promising solvation-docking hits had MM-PB/SA binding free energies better than 6.8 kcal/mol and the best one, HIT15, had -17.0 kcal/mol. In conclusion, the hierarchical multiple-filter database searching strategy is an attractive strategy in drug lead exploration. PMID- 15801835 TI - A new class of selective myocardial calcium channel modulators. 2. Role of the acetal chain in oxadiazol-3-one derivatives. AB - In the framework of the continuing interest of this research group in the use of 8-aryl-8-hydroxy-8H-[1,4]thiazino[3,4-c][1,2,4]oxadiazol-3-ones (1) as calcium entry blockers, a number of acetals were synthesized and assayed "in vitro". All of them are structurally related to diltiazem and pyrrolobenzothiazines. The effect on the biological profile was measured by functional assays for a wide variety of acetal residues: saturated linear and branched chains, short and long unsaturated E and/or Z chains as well as benzyl and methylcyclohexyl residues. From selective assays on the most active derivative (5b) (EC(50) = 0.04 microM), which is 20 times more active than diltiazem (EC(50) = 0.79 microM), a muscarinic or adenosinic mechanism of action was excluded. A 3D QSAR model was obtained and validated with homologous literature data, and a virtual receptor scheme was derived for the unknown binding site. The following pharmacophoric features favorably affect the potency: one positively charged center, three lipophilic groups, and two hydrogen-bonding acceptor groups. PMID- 15801836 TI - LEA3D: a computer-aided ligand design for structure-based drug design. AB - We present an improved version of the program LEA developed to design organic molecules. Rational drug design involves finding solutions to large combinatorial problems for which an exhaustive search is impractical. Genetic algorithms provide a tool for the investigation of such problems. New software, called LEA3D, is now able to conceive organic molecules by combining 3D fragments. Fragments were extracted from both biological compounds and known drugs. A fitness function guides the search process in optimizing the molecules toward an optimal value of the properties. The fitness function is build up by combining several independent property evaluations, including the score provided by the FlexX docking program. One application in de novo drug design is described. The example makes use of the structure of Mycobacterium tuberculosis thymidine monophosphate kinase to generate analogues of one of its natural substrates. Among 22 tested compounds, 17 show inhibitory activity in the micromolar range. PMID- 15801837 TI - Predicting human serum albumin affinity of interleukin-8 (CXCL8) inhibitors by 3D QSPR approach. AB - A novel class of 2-(R)-phenylpropionamides has been recently reported to inhibit in vitro and in vivo interleukin-8 (CXCL8)-induced biological activities. These CXCL8 inhibitors are derivatives of phenylpropionic nonsteroidal antiinflammatory drugs (NSAIDs), high-affinity ligands for site II of human serum albumin (HSA). Up to date, only a limited number of in silico models for the prediction of albumin protein binding are available. A three-dimensional quantitative structure property relationship (3D-QSPR) approach was used to model the experimental affinity constant (K(i)) to plasma proteins of 37 structurally related molecules, using physicochemical and 3D-pharmacophoric descriptors. Molecular docking studies highlighted that training set molecules preferentially bind site II of HSA. The obtained model shows satisfactory statistical parameters both in fitting and predicting validation. External validation confirmed the statistical significance of the chemometric model, which is a powerful tool for the prediction of HSA binding in virtual libraries of structurally related compounds. PMID- 15801838 TI - Architecture of the human urotensin II receptor: comparison of the binding domains of peptide and non-peptide urotensin II agonists. AB - The human urotensin II receptor (h-UTR) is a member of the family of rhodopsin like G-protein-coupled receptors (GPCRs) involved in the modulation of the functionality of many tissues and organs. Recently the urotensin-II (UII) neuropeptide, which is a potent vasoconstrictor in mammals and it is postulated to play a central role in cardiovascular homeostasis, has been identified as an agonist of the UII receptor. To elucidate the receptor's molecular recognition, a h-UTR model was constructed by homology modeling using the 2.6 A crystal structure of bovine rhodopsin as a template and subsequently refined by molecular dynamics simulations. The molecular recognition of h-UTR was probed by automated docking of P5U, a potent UII peptide agonist, as well as of the non-peptide compounds 1-4. We believe that this new model of the h-UTR provides the means for understanding the ligand's potency and for facilitating the design of novel and more potent UII ligands. PMID- 15801839 TI - CoMFA and CoMSIA investigations revealing novel insights into the binding modes of dopamine D3 receptor agonists. AB - As an extension of a series of dopamine D(3) receptor agonists involving FAUC 54, ex-chiral pool synthesis, and biological evaluation of 3-substituted 7 aminotetrahydroindolizines was performed. Considering the structural features of both series of enantiomers, we developed a novel alignment hypothesis for D(3) agonists, allowing for the placement of the aromatic moieties on two alternative, adjacent positions. CoMFA and CoMSIA analyses yielded significant cross-validated q(2) values of 0.726 and 0.590, respectively, when a newly invented program application (IRAS) controlling the alignment selection proved to be useful. Employing the CoMFA/CoMSIA contribution maps, we were able to transform a previously constructed homology model of the D(3) receptor from an inactive into an activate state. Besides the established ionic interactions, we propose pi stacking with Phe6.51 and a hydrogen bond between His6.55 and the acyl moiety to be primarily involved in the D(3) receptor binding of FAUC 54 and its analogues. PMID- 15801840 TI - Substituted 5,5'-diphenyl-2-thioxoimidazolidin-4-one as CB1 cannabinoid receptor ligands: synthesis and pharmacological evaluation. AB - A set of 30 substituted 5,5'-diphenyl-2-thioxoimidazolidin-4-one (thiohydantoins) derivatives was synthesized, and their affinity for the human CB(1) cannabinoid receptor has been evaluated. These compounds are derived from the previously described cannabinoid ligands 5,5'-diphenylimidazolidine-2,4-dione (hydantoins). The replacement of the oxygen by a sulfur leads to an increase of the affinity while the function-i.e., inverse agonism-determined by [(35)S]GTPgammaS experiments remains unaffected. Finally, to evaluate the molecular parameters that could influence the affinity of the thiohydantoins, molecular electrostatic potential as well as lipophilicity calculations were undertaken on representative thiohydantoins and hydantoins derivatives. In conclusion, 5,5'-bis-(4-iodophenyl) 3-butyl-2-thioxoimidazolidin-4-one (31) and 3-allyl-5,5'-bis(4-bromophenyl)-2 thioxoimidazolidin-4-one (32) possess the highest affinity for the CB(1) cannabinoid receptor described to date for the hydantoin and thiohydantoins series when compared in a same bioassay. PMID- 15801841 TI - Druggability indices for protein targets derived from NMR-based screening data. AB - An analysis of heteronuclear-NMR-based screening data is used to derive relationships between the ability of small molecules to bind to a protein and various parameters that describe the protein binding site. It is found that a simple model including terms for polar and apolar surface area, surface complexity, and pocket dimensions accurately predicts the experimental screening hit rates with an R(2) of 0.72, an adjusted R(2) of 0.65, and a leave-one-out Q(2) of 0.56. Application of the model to predict the druggability of protein targets not used in the training set correctly classified 94% of the proteins for which high-affinity, noncovalent, druglike leads have been reported. In addition to understanding the pocket characteristics that contribute to high-affinity binding, the relationships that have been defined allow for quantitative comparative analyses of protein binding sites for use in target assessment and validation, virtual ligand screening, and structure-based drug design. PMID- 15801842 TI - Identification of 1,2,3,4,5,6-hexabromocyclohexane as a small molecule inhibitor of jak2 tyrosine kinase autophosphorylation [correction of autophophorylation]. AB - The commercially available Jak2 inhibitor, alpha-cyano-3,4-dihydroxy-N benzylcinnamide (AG490), has been used extensively to study Jak2 kinase function. While alpha-cyano-3,4-dihydroxy-N-benzylcinnamide is a potent Jak2 inhibitor, it can inhibit a number of other kinase signaling pathways as well. To circumvent this problem, we sought to identify novel small molecule inhibitors of Jak2 tyrosine kinase activity. For this, we constructed a homology model of the Jak2 kinase domain and identified solvent accessible pockets on the surface of the structure. Using the DOCK program, we tested 6451 compounds of known chemical structure in silico for their ability to interact with a pocket positioned adjacent to the activation loop. We attained the top seven scoring compounds from the National Cancer Institute and tested their ability to inhibit Jak2 autophosphorylation in vitro. Using Western blot analysis, we found that one of the compounds, 1,2,3,4,5,6-hexabromocyclohexane, was able to potently, and directly, inhibit Jak2 autophosphorylation. Characterization of this compound revealed that it inhibits Jak2 tyrosine autophosphorylation in both a time- and concentration-dependent manner. It greatly reduced growth hormone-mediated Jak2 autophosphorylation but did not block autophosphorylation of the epidermal growth factor receptor. Furthermore, doses as high as 100 muM were not toxic to cells as measured by their ability to exclude propidium iodide. As such, we believe that this compound could serve as a lead compound for a new generation of Jak2 inhibitors and, perhaps, be useful in elucidating the mechanisms of Jak2 kinase function. PMID- 15801843 TI - Virtual screening workflow development guided by the "receiver operating characteristic" curve approach. Application to high-throughput docking on metabotropic glutamate receptor subtype 4. AB - The "receiver operating characteristic" (ROC) curve method is a well-recognized metric used as an objective way to evaluate the ability of a given test to discriminate between two populations. This facilitates decision-making in a plethora of fields in which a wrong judgment may have serious consequences including clinical diagnosis, public safety, travel security, and economic strategies. When virtual screening is used to speed-up the drug discovery process in pharmaceutical research, taking the right decision upon selecting or discarding a molecule prior to in vitro evaluation is of paramount importance. Characterizing both the ability of a virtual screening workflow to select active molecules and the ability to discard inactive ones, the ROC curve approach is well suited for this critical decision gate. As a case study, the first virtual screening workflow focused on metabotropic glutamate receptor subtype 4 (mGlu4R) agonists is reported here. Six compounds out of 38 selected and tested in vitro were shown to have agonist activity on this target of therapeutic interest. PMID- 15801844 TI - Synthesis and structure-activity relationships of a new model of arylpiperazines. 8. Computational simulation of ligand-receptor interaction of 5-HT(1A)R agonists with selectivity over alpha1-adrenoceptors. AB - We have designed and synthesized a new series of arylpiperazines V exhibiting high 5-HT(1A)R affinity and selectivity over alpha(1)-adrenoceptors. The new selective 5-HT(1A)R ligands contain a hydantoin (m = 0) or diketopiperazine (m = 1) moiety and an arylpiperazine moiety separated by one methylene unit (n = 1). The aryl substituent of the piperazine moiety (Ar) consists of different benzofused rings mimicking the favorable voluminous substituents at ortho and meta positions predicted by 3D-QSAR analysis in the previously reported series I. In particular, (S)-2-[[4-(naphth-1-yl)piperazin-1-yl]methyl]-1,4 dioxoperhydropyrrolo[1,2-a]pyrazine [(S)-9, CSP-2503] (5-HT(1A), K(i) = 4.1 nM; alpha(1), K(i) > 1000 nM) has been pharmacologically characterized as a 5-HT(1A)R agonist at somatodendritic and postsynaptic sites, endowed with anxiolytic properties. Ligand (S)-9 is predicted, in computer simulations, to bind Asp(3.32) in TMH 3, Thr(5.39) and Ser(5.42) in TMH 5, and Trp(6.48) in TMH 6. We propose that agonists modify, by means of an explicit hydrogen bond, the conformation of Trp(6.48) from pointing toward TMH 7, in the inactive gauche+ conformation, to pointing toward the ligand binding site, in the active trans conformation. PMID- 15801845 TI - Fluorinated diaryl sulfides as serotonin transporter ligands: synthesis, structure-activity relationship study, and in vivo evaluation of fluorine-18 labeled compounds as PET imaging agents. AB - A series of new, fluorine-containing substituted diphenyl sulfides was synthesized to serve as candidate ligands for positron emission tomography (PET) imaging of the serotonin transporter (SERT) and to further probe the structure activity relationship (SAR) of this class of compounds. Candidate compounds were assayed for their affinities to the monoamine transporters (SERT, norepinephrine transporter (NET), and dopamine transporter (DAT)) in competitive binding experiments in vitro using cloned human transporters. From these in vitro assays, four compounds (7c-f) were chosen for further evaluation. All four compounds have nanomolar affinity for SERT (K(i) 1.46 nM, 1.04 nM,1.83 nM, and 3.58 nM for 7c, 7d, 7e, and 7f, respectively). The F-18-labeled compounds, 16 and 18a-c, were prepared via a two-step radiosynthesis. Biodistribution studies in rats indicated that the F-18-labeled compounds localized in brain regions with high concentrations of SERT. Furthermore, competition experiments demonstrated that the binding of these radioligands in the rat brain was saturable, specific, and selective to SERT. Specific binding in the rat hypothalamus peaked at 5.6 for ligand 16 and 4.4 for 18b at 90 min after radioactivity administration. For ligand 18a, this same ratio was 8.4 at 120 min postinjection, while compound 18c displayed a lower specific binding ratio of 2.4. In summary, four F-18-labeled ligands were prepared and evaluated as candidate PET imaging agents for SERT. Among these four ligands, three appear to be promising radioligands suitable for the labeling of SERT in vivo, with 18a providing a higher specific binding in vivo than 16 or 18b. PMID- 15801846 TI - High-throughput lipophilicity measurement with immobilized artificial membranes. AB - We report on a new, high-throughput assay designed to measure octanol/water partition coefficients in early drug discovery. The assay is carried out in 96 well microtiterplates and measures the diffusion of compounds between two aqueous compartments separated by a thin octanol liquid layer. Octanol/water partition coefficients are derived from the apparent permeability (P(a)) values using a calibration curve. The assay can measure partition coefficients within the range 2 to + 8; thus, a dynamic range of 10 log units can be covered in one single run. Unlike chromatographic methods, the technology is not restricted to neutral and weakly basic compounds, and, as no stationary phase is involved, the data can be strictly compared with values obtained from traditional methods such as shake flask/HPLC or dual-phase potentiometric titration. PMID- 15801847 TI - N-[18F] fluoroethylpiperidin-4ylmethyl acetate, a novel lipophilic acetylcholine analogue for PET measurement of brain acetylcholinesterase activity. AB - The reduction of acetylcholinesterase (AChE) activity in the brain has been measured in dementia disorders such as Alzheimer's disease and dementia with Lewy bodies using (11)C-labeled acetylcholine analogues, N-[(11)C]methylpiperidin-4-yl acetate and propionate, and positron emission tomography (PET). Our aim was to develop an (18)F-labeled acetylcholine analogue useful for brain AChE mapping with PET, since (18)F, with a longer half-life, has advantages over (11)C. In a preliminary study, a series of N-[(14)C]ethylpiperidin-3-yl or -4-ylmethanol esters (acetyl and propionyl esters) were newly designed and evaluated in vitro regarding the reactivity with and specificity to AChE using purified human enzymes, leading to a novel (18)F-labeled acetylcholine analogue, N [(18)F]fluoroethylpiperidin-4-ylmethyl acetate. In rat experiments, the (18)F labeled candidate showed desirable properties for PET AChE measurement: high brain uptake of the authentic ester, high AChE specificity, a moderate hydrolysis rate, and low membrane permeability (metabolic trapping) of the metabolite. PMID- 15801848 TI - Discovery of 2-arylthiazolidine-4-carboxylic acid amides as a new class of cytotoxic agents for prostate cancer. AB - To improve the selectivity and antiproliferative activity of previously reported serine amide phosphates (SAPs), we designed a new series of 4-thiazolidinone amides, in which the 4-thiazolidinone moiety was introduced as a phosphate mimic. However, these 4-thiazolidinone derivatives demonstrated less cytotoxicity in prostate cancer cells despite improved selectivity over RH7777 cells. To further optimize the thiazolidinone analogues in terms of cytotoxicity and selectivity, we made closely related structural modifications, which led us to the discovery of a new class of 2-arylthiazolidine-4-carboxylic acid amides. These compounds were potent cytotoxic agents with IC(50) values in the low micromolar concentration range and demonstrated enhanced selectivity in receptor-negative cells compared to SAPs and 4-thiazolidinone amides. PMID- 15801849 TI - Lipopolysaccharide sequestrants: structural correlates of activity and toxicity in novel acylhomospermines. AB - Lipopolysaccharides (LPS), otherwise termed "endotoxins", are outer membrane constituents of Gram-negative bacteria. Lipopolysaccharides play a key role in the pathogenesis of "septic shock", a major cause of mortality in the critically ill patient. Therapeutic options aimed at limiting downstream systemic inflammatory processes by targeting lipopolysaccharide do not exist at the present time. We have defined the pharmacophore necessary for small molecules to specifically bind and neutralize LPS and, using animal models of sepsis, have shown that the sequestration of circulatory LPS by small molecules is a therapeutically viable strategy. In this paper, the interactions of a series of acylated homologated spermine compounds with LPS have been characterized. The optimal acyl chain length for effective sequestration of LPS was identified to be C(16) for the monoacyl compounds. The most promising of these compounds, 4e, binds LPS with an ED(50) of 1.37 muM. Nitric oxide production in murine J774A.1 cells, as well as TNF-alpha in human blood, is inhibited in a dose-dependent manner by 4e at concentrations orders of magnitude lower than toxic doses. Administration of 4e to d-galactosamine-sensitized mice challenged with supralethal doses of LPS provided significant protection against lethality. Potent antiendotoxic activity, low toxicity, and ease of synthesis render this class of compounds candidate endotoxin-sequestering agents of potential significant therapeutic value. PMID- 15801850 TI - Syntheses and biological activities of rebeccamycin analogues with uncommon sugars. AB - Rebeccamycin analogues containing uncommon sugars and substitutions on the imide nitrogen have been synthesized. Their cytotoxicities were tested in colon cancer and leukemia cells. Their ability to target topoisomerase I was examined using the in vivo complex of the topoisomerase bioassay in Hela cells. Compared with aglycon 1, the modified compounds with various sugar moieties showed more potent cytotoxicities and topo I targeting ability. In addition, the rebeccamycin analogues with various uncommon sugars showed distinct cytotoxicities and topo I targeting activities. The activity of compounds with 2-deoxyglucose (8 and 9) > compounds with 2,6-deoxyglucose (5 and 6) > compounds with 2,3,6-deoxyglucose (10). Furthermore, the anticancer activity of compounds correlated with their ability to target endogenous topo I. These results suggest that the sugar moiety, especially the 2-OH and 6-OH group of the sugar, rather than the modifications in imide structure on the indolocarbazole ring, is a key element for its activity. PMID- 15801851 TI - From 1-acyl-beta-lactam human cytomegalovirus protease inhibitors to 1 benzyloxycarbonylazetidines with improved antiviral activity. A straightforward approach to convert covalent to noncovalent inhibitors. AB - Starting from the structure of known beta-lactam covalent human cytomegalovirus (HCMV) protease inhibitors and from the knowledge of the residues implicated in the active site of this enzyme, we designed a series of phenylalanine-derived 2 azetidinones bearing a 4-carboxylate moiety that could be apt for additional interactions with the guanidine group of the Arg165/Arg166 residues of the viral protease. Some compounds within this series showed anti-HCMV activity at 10-50 muM, but rather high toxicity. The presence of aromatic 1-acyl groups and a certain hydrophobic character in the region of the 4-carboxylate were stringent requirements for anti-HCMV activity. To go a step ahead into the search for effective HCMV medicines, we then envisaged a series of noncovalent inhibitors by simple deletion of the carbonyl group in the beta-lactam derivatives to provide the corresponding azetidines. This led to low micromolar inhibitors of HCMV replication, with 17 and 27 being particularly promising lead compounds for further investigation, although their toxicity still needs to be lowered. PMID- 15801852 TI - Sodium nitroprusside: mechanism of NO release mediated by sulfhydryl-containing molecules. AB - Sodium nitroprusside (SNP) is among the most widely studied nitric oxide donors, and its capability of producing NO seems to depend on its interaction with sulfhydryl-containing molecules present in vivo. The aim of this research has been the study of the mechanism of interaction between SNP and sulfhydryl containing compounds, such as cysteine and glutathione, through detection by EPR, UV-vis, and IR spectroscopy of both the radical and nonradical species involved. An electron-transfer process can be invoked as the key step, which leads to the formation of the reduced SNP radical, the main detectable radical intermediate, and the corresponding S-nitrosothiol, the ending product of NO that can be considered the real storage and transporters of NO. When cysteine was used, a second radical species (A) is detectable: it can be accounted for by the interaction of a byproduct with unreacted cysteine. PMID- 15801853 TI - Synthesis and pharmacology of N1-substituted piperazine-2,3-dicarboxylic acid derivatives acting as NMDA receptor antagonists. AB - The binding site for competitive NMDA receptor antagonists is on the NR2 subunit, of which there are four types (NR2A-D). Typical antagonists such as (R)-AP5 have a subunit selectivity of NR2A > NR2B > NR2C > NR2D. The competitive NMDA receptor antagonist (2R,3S)-(1-biphenylyl-4-carbonyl)piperazine-2,3-dicarboxylic acid (PBPD, 16b) displays an unusual selectivity with improved relative affinity for NR2C and NR2D vs NR2A and NR2B. Analogues of 16b bearing aroyl or aryl substituents attached to the N(1) position of piperazine-2,3-dicarboxylic acid have been synthesized to probe the structural requirements for NR2C/NR2D selectivity. A phenanthrenyl-2-carbonyl analogue, 16e, had >60-fold higher affinity for NR2C and NR2D and showed 3-5-fold selectivity for NR2C/NR2D vs NR2A/NR2B. The phenanthrenyl-3-carbonyl analogue (16f) was less potent but more selective, having 5- and 7-fold selectivity for NR2D vs NR2A and NR2B, respectively. Thus, antagonists bearing bulky hydrophobic residues have a different NR2 subunit selectivity than that of typical antagonists. PMID- 15801854 TI - Synthesis, structure-activity relationships at the GABA(A) receptor in rat brain, and differential electrophysiological profile at the recombinant human GABA(A) receptor of a series of substituted 1,2-diphenylimidazoles. AB - A series of new 1,2-diphenylimidazole derivatives (1a-x) were synthesized and evaluated for their ability to potentiate gamma-aminobutyric acid (GABA)-evoked currents in Xenopus laevis oocytes expressing recombinant human GABA(A) receptors. Many of these compounds enhanced GABA action with potencies (EC(50) = 0.19-19 muM) and efficacies (maximal efficacies of up to 640%) similar to or greater than those of anesthetics such as etomidate, propofol, and alphaxalone. Structure-activity relationship analysis revealed that the presence of an ester moiety in the imidazole ring was required for full agonist properties, while modifications made in the phenyl rings affected potency and efficacy, with ethyl 2-(4-bromophenyl)-1-(2,4-dichlorophenyl)-1H-4-imidazolecarboxylate showing the highest potency. These compounds potentiated the [(3)H]GABA binding to rat brain membranes, suggesting a site of interaction different from that of GABA. As for etomidate, mutation of asparagine-265 in the beta2 subunit of the GABA(A) receptor into serine reduced the ability of derivative 1i to modulate the GABA function. PMID- 15801855 TI - Preparation and pharmacological characterization of trans-2-amino-5(6)-fluoro 6(5)-hydroxy-1-phenyl-2,3-dihydro-1H-indenes as D2-like dopamine receptor agonists. AB - The present work reports the synthesis of trans-2-amino-5(6)-fluoro-6(5)-hydroxy 1-phenyl-2,3-dihydro-1H-indenes (4a-f, 5a-f) as a continuation of our studies to better understand the significance of the halo substituent in the trans-1-phenyl 2-aminoindane series and to extend knowledge of the monophenolic ligands of DA receptors. The affinity of the new compounds and related methoxylated precursors (10-15 and 18-23) was estimated in vitro by displacement of [(3)H]SCH23390 (for D(1)-like receptors) or [(3)H]YM-09-151-2 (for D(2)-like receptors) from homogenates of porcine striatal membranes. The results indicate that unsubstituted amines 4a, 5a, 10, and 11 are poorly effective at DA receptors. The introduction of two n-propyl groups on the nitrogen atom (compounds 14, 15, 4c, and 5c) and N-allyl-N-methyl- or N-methyl-N-propyl- substitution (compounds 20 23, 4e, 4f, 5e, 5f) increased the D(2)-like affinities and selectivity. The D(2) like agonistic activity of selected compounds 15, 20, 21, 4e, 5c, and 5e was proved by evaluating their effects on the cyclic guanosine monophosphate (cGMP) content in rat neostriatal membranes. All tested compounds displayed a potential dopamine D(2)-like agonist profile decreasing basal levels of cGMP. The selective D(2)-like agonism of compounds 20 and 5e was proved by their effects on basal striatal adenylyl cyclase activity. PMID- 15801857 TI - Cationic chalcone antibiotics. Design, synthesis, and mechanism of action. AB - This paper describes how the introduction of "cationic" aliphatic amino groups in the chalcone scaffold results in potent antibacterial compounds. It is shown that the most favorable position for the aliphatic amino group is the 2-position of the B-ring, in particular in combination with a lipophilic substituent in the 5 position of the B-ring. We demonstrate that the compounds act by unselective disruption of cell membranes. Introduction of an additional aliphatic amino group in the A-ring results in compounds that are selective for bacterial membranes combined with a high antibacterial activity against both Gram-positive and negative pathogens. The most potent compound in this study (78) has an MIC value of 2 muM against methicillin resistant Staphylococus aureus. PMID- 15801856 TI - No auxiliary, no byproduct strategy for water-soluble prodrugs of taxoids: scope and limitation of O-N intramolecular acyl and acyloxy migration reactions. AB - Since numerous new taxoids active against multidrug resistant (MDR) tumors have been developed and their poor water-solubility is a very real problem in intravenous administration, we have designed and synthesized a series of novel water-soluble taxoid prodrugs (isotaxoids). These prodrugs, a 2'-O-isoform of taxoids, showed promising results with higher water solubility (0.8-1.1 mg/mL) and proper kinetics for parent drug release by a simple pH-dependent chemical mechanism via O-N intramolecular acyl migration. No additional functional auxiliaries are released during the conversion to parent drugs, which would be an advantage in toxicology and general pharmacology, and the cost for the evaluations of auxiliary units in these fields could be saved in prodrug development. In addition, we demonstrate for the first time the successful application of the O-N intramolecular acyloxy migration reaction in the prodrug design, with the exception of the tert-butyloxycarbonyl group, and that this reaction can be provided with no organic solvent and no side products. PMID- 15801858 TI - Discovery of the alpha7 nicotinic acetylcholine receptor agonists. (R)-3'-(5 Chlorothiophen-2-yl)spiro-1-azabicyclo[2.2.2]octane-3,5'-[1',3']oxazolidin-2'-one as a novel, potent, selective, and orally bioavailable ligand. AB - Recent advances in molecular biology suggest that neuronal nicotinic acetylcholine receptors play important roles in the central nervous system (CNS). Of these receptors, the alpha7 group has recently attracted interest for its CNS related actions and is looked to as a potential new class of pharmacological targets for cognition, schizophrenia, sensory gating, and anxiety. In the course of a research program aimed at the discovery of alpha7 receptor agonists with high affinity, subtype selectivity, and good pharmacokinetic profile, we discovered (R)-3'-(5-chlorothiophen-2-yl)spiro-1-azabicyclo[2.2.2]octane-3,5' [1',3']oxazolidin-2'-one (25). Compound 25 has potent binding affinity (K(i) = 9 nmol/L) and good selectivity toward the other nicotinic subtypes (alpha4beta2 and alpha1beta2gammadelta) and has been found in pharmacokinetic evaluation to have good oral bioavailability and brain permeability. PMID- 15801859 TI - Anchor-GRIND: filling the gap between standard 3D QSAR and the GRid-INdependent descriptors. AB - The aim of this work is to present the anchor-GRIND methodology. Anchor-GRIND efficiently combines a priori chemical and biological knowledge about the studied compounds with alignment-independent molecular descriptors derived from molecular interaction fields. Such descriptors are particularly useful for series of ligands sharing a common scaffold but with very diverse substituents. The method uses a specific position of the molecular structure (the "anchor point") to compare the spatial distribution of the molecular interaction fields of the substituents. The descriptors produced are more detailed and specific than the original GRIND while still avoiding the bias introduced by the alignment. Three data sets have been studied to demonstrate the usefulness of the anchor-GRIND methodology for 3D-QSAR modeling. The two first data sets respectively include congeneric series of the hepatitis C virus NS3 protease and of the acetylcholinesterase inhibitors. The third data set discriminates between factor Xa inhibitors of high and low affinity. In all the series presented, the models obtained with the anchor-GRIND are statistically sound and easy to interpret. PMID- 15801860 TI - Synthesis of 2',3'-dideoxynucleoside 5'-alpha-P-borano-beta,gamma (difluoromethylene)triphosphates and their inhibition of HIV-1 reverse transcriptase. AB - The triphosphates of antiviral 2',3'-dideoxynucleosides (ddNs) are the active chemical species that inhibit viral DNA synthesis. The inhibition involves incorporation of ddNMP into DNA and subsequent chain termination. A conceivable strategy for antiviral drugs is to employ nucleoside 5'-triphosphate mimics that can entirely bypass cellular phosphorylation. AZT 5'-alpha-R(P)-borano-beta,gamma (difluoromethylene)triphosphate (5'-alphaB-betagammaCF(2)TP) has been identified as a potent inhibitor of HIV-1 reverse transcriptase (HIV-1 RT). This work was aimed at confirming that 5'-alphaB-betagammaCF(2)TP is a useful generic triphosphate moiety and can render antiviral ddNs with potent inhibitory effects on HIV-1 RT. Thus, 10 ddNs were converted to their 5'-alphaB-betagammaCF(2)TPs via a sequence (one-pot) of reactions: formation of an activated phosphite, formation of a cyclic triphosphate, boronation, and hydrolysis. Other synthetic routes were also explored. All ddN 5'-alphaB-betagammaCF(2)TPs tested exhibited essentially the same level of inhibition of HIV-1 RT as the corresponding ddNTPs. A conclusion can be made that 5'-alphaB-betagammaCF(2)TP is a generic and promising triphosphate mimic (P3M) concerning HIV-1 RT inhibition and serum stability. It is anticipated that use of 5'-alphaB-betagammaCF(2)TP as P3M moiety will lead to the discovery of a new class of anti-HIV agents. PMID- 15801861 TI - Synthesis of some cryptolepine analogues, assessment of their antimalarial and cytotoxic activities, and consideration of their antimalarial mode of action. AB - A series of analogues of cryptolepine (1) have been synthesized and evaluated for their in vitro antiplasmodial and cytotoxic properties. The IC(50) values of several compounds (11a, 11k-m, 11o, 13) against Plasmodium falciparum (strain K1) were <0.1 muM, 5-10-fold lower than that of 1 but their cytotoxicities were only 2-4 times greater than that of 1. Compounds with a halogen in the quinoline ring and a halogen or a nitro group in the indole ring have enhanced antiplasmodial activity. In mice infected with P. berghei, the 7-bromo-2-chloro (11k) and 2 bromo-7-nitro (13) derivatives of 1 suppressed parasitemia by >90% at doses of 25 mg kg(-1) day(-1) with no apparent toxicity to the mice. 2,7-Dibromocryptolepine (15) was evaluated at several dose levels, and a dose-dependent suppression of parasitemia was seen (ED(90) = 21.6 mg kg(-1) day(-1)). The antimalarial mode of action of 1 appears to be similar to that of chloroquine and involves the inhibition of hemozoin formation. A number of analogues were assessed for their effects on the inhibition of beta-hematin (hemozoin) formation, and the results were compared with their antiplasmodial activities having taken account of their predicted accumulation into the acidic parasite food vacuole. No correlation was seen (r(2) = 0.0781) suggesting that the potent antimalarial activity of compounds such as 15 involves other mechanisms in addition to the inhibition of hemozoin formation. PMID- 15801862 TI - Synthesis, biological activity, and SAR of antimycobacterial 9-aryl-, 9 arylsulfonyl-, and 9-benzyl-6-(2-furyl)purines. AB - 9-Aryl-, 9-arylsulfonyl- and 9-benzyl-6-(2-furyl)purines were synthesized by N alkylation or N-arylation of the purine followed by Stille coupling to introduce the furyl substituent in the 6-position and the compounds screened for activity against Mycobacterium tuberculosis. The 9-aryl- and 9-sulfonylarylpurines exhibited weak activity toward the bacteria, but 9-benzylpurines were good inhibitors especially those carrying electron-donating substituents on the phenyl ring. A chlorine atom in the purine 2-position further enhanced activity. The high antimycobacterial activity (MIC 0.39 microg/mL against M. tuberculosis), low toxicity against mammalian cells and activity inside macrophages found for 2 chloro-6-(2-furyl)-9-(4-methoxyphenylmethyl)-9H-purine makes this compound a highly interesting potential antituberculosis drug. PMID- 15801863 TI - Correlation between seminal carnitine and functional spermatozoal characteristics in men with semen dysfunction of various origins. AB - BACKGROUND AND OBJECTIVE: L-carnitine is an essential molecule involved in mitochondrial metabolism, controlling the transport of acetyl and acyl groups across the mitochondrial inner membrane. Carnitine and acetylated carnitine (L acetylcarnitine) are found in high concentrations in the epididymis, where they also act as antioxidants, protecting spermatozoa against damage caused by reactive oxygen species. In this open study we investigated the correlation between seminal carnitine levels and spermatozoal function, and the effect of combined L-carnitine + L-acetylcarnitine therapy, in infertile men. PATIENTS AND METHODS: 170 infertile men were enrolled in this study. Patients were divided into those with a total sperm motility below the normal WHO range (<50% motility, group 1 [n = 102]) and those with total sperm motility within the normal range (> or =50% motility, group 2 [n = 68]). Patients in group 1 were further divided into two groups: those with primary or secondary azoospermia (1B [n = 36]), and those without (1A [n = 66]). Patients in group 1A received L-carnitine 1 g/day and L-acetylcarnitine 500 mg twice daily for 6 months. Seminal carnitine levels were measured and correlated with sperm count and motility, eosin test, hypo osmotic swelling test, acridine orange test for sperm nuclear DNA integrity and sperm kinetics evaluated by computer-assisted sperm analysis in all patients. RESULTS: There was a significant correlation between seminal carnitine concentration and sperm concentration, total sperm count, sperm total motility, rapid forward progression, live sperm count, membrane function, nuclear DNA integrity, capacity for cervical mucus penetration, linearity of spermatic movement, and amplitude of lateral sperm head movement (all p < 0.0001) in the entire study population. In group 1A, there was a significant increase in total motility, live sperm count, membrane integrity and linearity of spermatic movement after 3 and 6 months of L-carnitine/L-acetylcarnitine treatment, and in capacity for cervical mucus penetration after 6 months of treatment, compared with baseline. CONCLUSION: Seminal carnitine concentration may be an appropriate marker of sperm and epididymal function. L-carnitine/L-acetylcarnitine treatment may be an effective therapy to improve mainly functional seminal parameters. PMID- 15801864 TI - Utilisation of human pharmacology studies with biomarkers for new drug applications in Japan. AB - OBJECTIVE: This study evaluated the utilisation of human pharmacology studies with biomarkers for either efficacy or safety estimation conducted for new drug applications (NDAs) submitted to the Japanese regulatory authority, the Ministry of Health, Labour and Welfare (MHLW). METHODS: A total of 50 new chemical entities (NCEs) posted on the Websites, which were approved from June 2000 to November 2001, were evaluated by investigating their approval information. The utilisation of human pharmacology studies with biomarkers was evaluated by focusing on the classification referred to biomarkers for either efficacy or safety estimation and timing of studies. RESULTS: The human pharmacology studies with biomarkers for either efficacy or safety estimation were conducted in 20 compounds classified by utilising measures of either efficacy (17 compounds) or safety (seven compounds). In 4 of 17 NCEs, some of the biomarkers in human pharmacology studies were similar to the clinical endpoints for efficacy assessment in therapeutic exploratory and/or therapeutic confirmatory studies. For safety assessment in therapeutic exploratory and/or therapeutic confirmatory studies, clinical endpoints rather than biomarkers in human pharmacology studies were used in all seven NCEs. The timing of each type of clinical study could only be obtained for 15 NCEs. Of these 15 NCEs, human pharmacology studies with biomarkers for either efficacy or safety estimation were conducted on six compounds. There were only two compounds for which human pharmacology studies with biomarkers for efficacy estimation were conducted before pivotal studies such as a therapeutic exploratory study or a bridging study. CONCLUSION: Our survey suggests that with Japanese NDAs, human pharmacology studies with biomarkers for either efficacy or safety estimation do not play a key role in accelerating drug development and maximising the knowledge gained from confirmatory trials. The relationship between a biomarker and a clinical endpoint should be investigated appropriately for accelerating drug development. We think that the utilisation of human pharmacology studies with biomarkers for either efficacy or safety estimation in the regulatory review process for NDAs should be encouraged with the advancements of drug evaluation research using an appropriate biomarker based on clinical pharmacology. PMID- 15801865 TI - Erythrocyte glutathione levels in lithium-induced hypothyroidism. AB - BACKGROUND AND OBJECTIVE: Lithium, widely used in the prophylaxis of psychiatric patients with bipolar affective disorders, is well known to induce thyroid alterations. However, a possible metabolic linkage between blood thyroxine levels and its regulatory function on erythrocyte glutathione concentration has not yet been evaluated in lithium-treated patients. The aim of this study was to assess the antioxidative capacity of erythrocytes in lithium-induced hypothyroidism before and after thyroxine replacement. PATIENTS AND METHODS: Thyroid ultrasound with clinical and laboratory evaluation of thyroid function and thyroid stimulating hormone assay were performed prior to and at 6-month intervals during lithium prophylaxis in 76 patients with bipolar affective disorders. The daily lithium dosage was adjusted to 600-1200 mg and the mean duration of treatment was 2.2 +/- 0.4 years. Final assessment revealed that 12 patients had evidence of primary hypothyroidism, and these were assigned as the test group. Lithium prophylaxis was supplemented with thyroxine at a dosage of 100 mg/day within 6 months after thyroid dysfunction was diagnosed. Red blood cell superoxide dismutase activities and the glutathione contents were measured before and after thyroxine replacement. In order to assess the effect of long-term lithium administration on red blood cell glutathione and superoxide dismutase levels, 12 patients who had not developed hypothyroidism during the follow-up period were selected for the lithium-treated euthyroid group. Mann Whitney U-test and Wilcoxon rank sum W-test were used for comparison of data. RESULTS: A comparison of the lithium-treated test group with healthy volunteers and their own values after thyroxine replacement revealed a significant decrease in red blood cell glutathione concentrations (p = 0.000) in the hypothyroid state. However, no clinically significant changes were observed in red blood cell superoxide dismutase activities of the test group. A statistical survey also demonstrated that there was no significant difference in the thyroid-stimulating hormone values as well as the red blood cell glutathione contents or superoxide dismutase activities between healthy controls and lithium-treated euthyroid subjects. CONCLUSIONS: It is most likely that lithium primarily inhibited hormone production in the thyroid and that this led to a compensatory increase in thyroid stimulating hormone secretion with a significant decrease in the red blood cell glutathione content. While the red blood cell glutathione content of hypothyroid patients was reduced to 40% of the post-thyroxine level, unchanged superoxide dismutase activity might render the erythrocytes vulnerable to oxidative stress and ultimately haemolysis. Thyroxine replacement during lithium prophylaxis of psychiatric patients is advisable in order to prevent subclinical hypothyroidism and related defects of erythrocyte antioxidant capacity. PMID- 15801866 TI - Pharmacokinetics and pilot efficacy of a mesalazine rectal gel in distal ulcerative colitis. PMID- 15801867 TI - Glufosfamide: beta-D-Glc-IPM, D 19575. AB - Glufosfamide [D 19575, beta-D-Glc-IPM] is a next-generation glucose conjugate of ifosfamide that is under development with Threshold Pharmaceuticals for the treatment of cancer. It is an alkylating agent in which isophosphoramide mustard, the alkylating metabolite of ifosfamide, is glycosidically linked to beta-D glucose. Cellular uptake of glufosfamide is mediated by a sodium-dependent transmembrane transporter protein of glucose and possibly also by other transporter proteins. Threshold is using its Metabolic Targeting technology to exploit unique aspects of tumour metabolism, particularly the elevated glucose utilisation of tumour cells to selectively target glufosfamide to the tumour site. Glufosfamide was originally developed from a research collaboration between Asta Medica (Degussa) and the Cancer Research Centre (DKFZ) in Heidelberg, Germany. In October 2001, Baxter International acquired the oncology division of ASTA Medica, and renamed it Baxter Oncology GmbH. According to its 2002 Annual Report, Baxter announced that it was terminating development of glufosfamide. Subsequently, Baxter and Threshold Pharmaceuticals entered into an exclusive licensing and development agreement in August 2003. Threshold has responsibility for the development and commercialisation of glufosfamide, primarily for use as an antitumour agent. In addition, Baxter manufactures glufosfamide on Threshold's behalf. Threshold received fast-track status for glufosfamide from the US FDA in the treatment of metastatic pancreatic cancer refractory to gemcitabine in November 2004. In December 2004, Threshold initiated a phase I/II trial (TH-CR 301 Study) investigating glufosfamide in combination with gemcitabine as a first line treatment of pancreatic cancer or advanced solid tumours. The phase I portion of the study may enroll up to 15 patients. The maximum tolerable dose combination determined will then be used in the phase 2 portion of the study. Up to 42 patients with advanced pancreatic cancer will be enrolled at various sites in the US, Latin America and Brazil.Previously, glufosfamide had been in phase II trials among patients with pancreatic carcinoma in Germany with Baxter Oncology and with the EORTC in the UK as well as Greece. However, development has been discontinued. PMID- 15801868 TI - Histrelin Hydrogel Implant--Valera: Histrelin implant, LHRH-Hydrogel implant, RL 0903, SPD 424. AB - Valera Pharmaceuticals, formerly Hydro Med Sciences, is developing a once-yearly Histrelin Hydrogel Implant [Histrelin implant, LHRH-Hydrogel implant, RL 0903, SPD 424, Vantas], a subcutaneous (s.c.) reservoir device capable of long-term delivery of histrelin at constant release rates for the treatment of prostate cancer. Histrelin is a luteinising hormone-releasing hormone agonist (LHRH). A different formulation of the LHRH implant is currently in development for the treatment of central precocious puberty (CPP). On 4 September 2003, Hydro Med Sciences announced that it had changed its name to Valera Pharmaceuticals. Shire Pharmaceuticals had an option to market and distribute the histrelin implant outside the US, but in a realigned agreement announced in January 2002, Shire stated that HydroMed (now Valera) would be responsible for concluding the phase III studies, filing for regulatory approval and producing the implant, while also gaining marketing rights in the US. Shire has no further involvement in development, but retains an option to market and distribute the product outside the US. The product is available for rest-of-the-world licensing through Valera Pharmaceuticals' business development division. Paladin Labs has received the exclusive rights for the sale and marketing of histrelin hydrogel implant in Canada. Valera Pharmaceuticals will have the responsibility for manufacturing and completing development of the product. In July 2004, Paladin announced it had filed for regulatory approval with Health Canada for the treatment of prostate cancer. Phase III trials have been conducted and initially involved two open labelled, randomised, parallel studies that compared the hydrogel implant with the active comparators, leuprorelin acetate 22.5 mg depot (TAP Pharmaceutical's Lupron Depot) and a 3-month implant of goserelin acetate (Astra Zeneca's Zoladex). However, because of financial constraints, HydroMed discontinued recruitment in a phase II European study and could not rationalise keeping the comparator arm of the phase III study. The US FDA then gave permission to continue the US part of the programme without the comparator arm, but with appropriate increases in the patient sample size. Patient enrolment in the phase III trial (US and Canada) was completed in July 2002. Valera Pharmaceuticals believes the product may have advantages over standard prostate cancer treatments including reduced costly surgical procedures, lower dosing, increased patient compliance and peace-of-mind, as well as potentially less cost to the patient. The phase III studies were completed in the second half of 2003.CPP arises from the increased release by the pituitary gland hormone gonadotropins and is characterised by an early onset of sexual development in pre-adolescents. Currently, the treatment of CPP involves injections of synthetic gonadotropic hormone-releasing factor agonists such as Supprelin (histrelin acetate) and TAP Pharmaceutical's Lupron Depot-Red (leuprolide acetate) every 28 days, a potentially painful procedure. This treatment inhibits pituitary release of gonadotropins. Valera's histrelin implant is a compact, removable subcutaneous implant that can be applied under a local anaesthetic in doctor's surgery. PMID- 15801869 TI - Lasofoxifene: CP 336156, CP-336156. AB - Lasofoxifene [CP 336156] is a potent, nonsteroidal, tissue-selective estrogen receptor modulator (SERM). It has the bone-sparing and cardioprotective effects of estrogen, but lacks estrogen's uterine cancer risk. Lasofoxifene is under development with Ligand Pharmaceuticals and Pfizer (formerly Parke-Davis) for the prevention of postmenopausal osteoporosis and breast cancer. In June 2000, Parke Davis' parent company, Warner-Lambert, merged with Pfizer. The resulting company retained the Pfizer name and Parke-Davis was integrated into Pfizer Global Research and Development. The discovery of lasofoxifene resulted from a research collaboration between Pfizer and Ligand Pharmaceuticals. There was a contract dispute between the two companies relating to their research agreement. Under a settlement of litigation, Ligand is entitled to milestone and royalty payments. If Pfizer is successful in developing the drug through to regulatory approval in the US, Ligand could receive royalty revenues from lasofoxifene as early as 2003 2004. The royalties will be equal to 6% of net sales and will be in addition to milestone payments for continuing development of the drug. However, on 6 March 2002, Ligand Pharmaceutical announced an agreement with Royalty Pharma in which the latter purchased the rights to a share of these future payments. Under the agreement, Ligand received US dollars 6 million from Royalty Pharma in exchange for a 0.25% stake in net sales of three SERM products (lasofoxifene, bazedoxifene and bazedoxifene/Premarin) for a period of 10 years. Royalty Pharma retains the option to purchase, at escalating prices, additional rights (subject to timing restrictions) to extend this stake up to 1.0%, for a total of US dollars 56 million. In April 2002, Royalty Pharma exercised its first option to purchase an additional 0.125% of potential future sales of the three SERMS in exchange for US dollars 3 million. Subsequently, in December 2002, Royalty Pharma exercised an expanded option and agreed to pay Ligand US dollars 6.775 million for 0.1875% of potential future sales of SERM products. Royalty Pharma and Ligand Pharmaceutical amended their royalty agreement in October 2003 for the three SERM products. Under the amended agreement, Royalty Pharma exercised an option to pay Ligand US dollars 12.5 million, plus cumulative milestones of up to US dollars 2.5 million upon the launches of the three SERMs (provided they are approved by 30 September 2005), in exchange for 0.7% of potential future sales of the products for 10 years. In November 2004, Ligand Pharmaceuticals and Royalty Pharma further amended their existing royalty agreement for the three SERM products. Under the terms of the revised agreement, Royalty Pharma will purchase an additional 1.625% of the SERM products' net sales for US dollars 32.5 million, which represents an acceleration of the previous option timetable and an increase in the royalty amount as well as aggregate purchase price. Consequently, Royalty Pharma increased its rights to a total of 3.0125% of net sales of each SERM product for 10 years following the first commercial sale of each product and has no further options. Ligand retains an approximately equal portion of lasofoxifene and other SERM's net sales going forward and for periods that could exceed 10 years. The royalty rates owed to Royalty Pharma for the royalties just purchased could be reduced by one-third if product sales exceed certain thresholds. Payments from the royalty purchase are non-refundable, regardless of whether the products ever become successfully launched or not. Milestone payments owed by Ligand's partners as products achieve development and regulatory targets will be paid to Ligand as earned and are not included in this amended agreement. In September 2004, Ligand Pharmaceuticals earned a milestone payment of approximately US dollars 2 million from Pfizer, payable in 181,818 shares of Ligand stock held by Pfizer. The payment was triggered by Pfizer's NDA submission for lasofoxifene in August 2004. Under the terms of the agreement between Ligand and Pfizer, Ligand is entitled to receive an additional milestone upon successful approval of lasofoxifene. On 19 August 2004, Pfizer filed an NDA with the US FDA for lasofoxifene for the prevention of osteoporosis in postmenopausal women. Product launch is forecasted to occur in 2006-2007. Ligand reported in January 2004 at the 22nd Annual JP Morgan Healthcare Conference that it anticipated the availability of phase III data and NDA filing sometime in 2004. Lasofoxifene has undergone two phase III studies with Pfizer in the US as an orally administered therapy for postmenopausal osteoporosis. In June 2003, Pfizer reported that enrolment was completed in a trial evaluating lasofoxifene in the prevention of bone loss. The trial also evaluated lasofoxifene's effect on lipid levels. The trial enrolled approximately 2000 postmenopausal women. Another trial was conducted among 8500 patients to investigate lasofoxifene in the treatment of fractures. In addition, Pfizer began a third worldwide phase III trial to evaluate whether lasofoxifene reduced the risk of vertebral fractures, breast cancer and cardiovascular disease. At the 10th Annual Meeting of the Biotechnology Industry Organization (BIO-2003), Ligand also confirmed that lasofoxifene was in phase III development for breast cancer. Lasofoxifene is under clinical evaluation as a treatment for vaginal atrophy. According to Pfizer's pipeline in November 2004, the company anticipates regulatory submission for vaginal atrophy by the end of 2004. In June 2002, Ligand estimated that lasofoxifene has the potential to reach sales of US dollars 1-2 billion, pending approval. PMID- 15801870 TI - Promoting a healthy school environment. PMID- 15801871 TI - Arkansas school nurses' role in statewide assessment of body mass index to screen for overweight children and adolescents. AB - National surveys that have tracked weight and physical activity in the United States for more than 40 years have shown a continuing increase in the number of overweight children and adolescents. Overweight children and adolescents are showing an increase in diseases related to overweight: Type II diabetes, cardiovascular disease, hypertension, and orthopedic and respiratory conditions. Overweight children are also likely to become obese adults. This article describes Arkansas's efforts to connect with families through schools to prevent or reduce overweight in children. Arkansas school nurses measured the heights and weights of more than 400,000 public school students during the 2003-2004 school year. A detailed protocol for accurate measurement of students is provided. PMID- 15801872 TI - Improving the quality of students' dietary intake in the school setting. AB - The dramatic increase in our understanding of the brain's development throughout childhood has increased our knowledge of the significance of micronutrients, such as iron and vitamin B-12, for this development. Deficiencies of these micronutrients have been shown to have an impact on students' cognitive development. Regardless of this knowledge, students continue to make unhealthy food choices and develop poor dietary habits. School environments are places where there is an opportunity to practice healthy eating habits. Yet many school policies fail to address the sale of foods of minimal nutritional value in the school setting. School nurses can play a vital role in planning policies at the local and national level that support and encourage healthy food environments, performing assessments of the nutritional status of students, and spearheading the implementation of evidence-based health promotion programs. It is time for school nurses to take the lead in efforts aimed at improving the quality of students' dietary intake in the school setting. PMID- 15801873 TI - An integrative review of interventions for adolescent weight loss. AB - The number of overweight adolescents aged 12-19 has tripled during the past 2 decades. Although health risks associated with obesity in adolescence and adulthood are well documented in the literature, little is known about the efficacy of interventions to reduce health risks of this group. The purpose of this study was to conduct a systematic review to describe the scope, domain, and effectiveness of weight loss interventions with overweight adolescents. English language journal articles published in nursing, psychology, nutrition, medicine, and exercise physiology literature between 1980 and 2003 were retrieved. Seventeen studies using comparison or control groups and interventions directed at reductions in adolescent body mass index or body weight were identified. Descriptive findings of those studies are reported here. Five specific limitations of these studies emerged from the analysis of the interventions: the study findings have not been validated by replication, the samples failed to include adequate representation of Latino and African American male participants, family participation in studies has been inconsistent and infrequent, there is a need for attention to study dropout rates (with attrition reported as high as 45%), and there is a need for conceptual frameworks to guide the studies. PMID- 15801874 TI - Perceptions of school nurses regarding obesity in school-age children. AB - Obesity is epidemic in the nation's school-age population with African American and Hispanic children and adolescents specifically at risk. School nurses at elementary and middle public schools in the Missouri 8th Congressional District were surveyed regarding their perceptions of childhood obesity. School nurses supported preventive interventions and were knowledgeable of the risks of childhood obesity but were less accepting of treatment and counseling for obese children in the school setting. One third of the school nurses did not recommend treatment for weight loss, and half of the nurses provided overweight children with counseling and referral only when parents asked for help. Perceptions of school nurses regarding childhood obesity identified in this study included the following: (a) counseling for obesity is difficult, (b) parental support is lacking, and (c) competence in providing counseling is low. These perceptions are barriers to school nurses taking a more active role in the prevention and treatment of childhood obesity. PMID- 15801875 TI - Vending machine policies and practices in Delaware. AB - Overweight has reached alarming proportions among America's youth. Although the cause of the rise in overweight rates in children and adolescents is certainly the result of the interaction of a variety of factors, the presence of vending machines in schools is one issue that has recently come to the forefront. Many states have passed or proposed legislation that limits student access to vending machines in schools or require that vending machines in schools offer healthier choices. The purposes of this study were (a) to assess the food and beverage vending machine offerings in the public school districts in the state of Delaware and (b) to determine whether there are any district vending policies in place other than the current U.S. Department of Agriculture regulations. The results of this study indicate the most commonly sold food and drink items in school vending machines are of minimal nutritional value. School administrators are most frequently in charge of the vending contract, as well as setting and enforcing vending machine policies. Suggestions are offered to assist school nurses, often the only health professional in the school, in becoming advocates for changes in school vending practices and policies that promote the health and well-being of children and adolescents. PMID- 15801876 TI - The breakfast-eating habits of inner city high school students. AB - This cross-sectional, descriptive correlational research study describes the breakfast-eating habits of 846 inner-city high school students. Fifty-seven percent of students reported skipping breakfast on the day of the survey, despite the free hot-breakfast program at their high school. Significantly more girls than boys skipped breakfast, and 10th grade students had the highest rate of skipping breakfast. Sixty-four percent of breakfast-skippers cited a lack of time, and 28% stated they could not eat early in the morning. More breakfast eaters reported eating at home (48%); only 14% reported eating at school, with 3% reportedly eating both at home and school. Milk, orange juice, cereal, and foods in the bread group were the most frequently eaten foods. Patterns of eating by gender and by grade level are discussed in this article, as are implications for school nursing, including assessment, teaching, and research. It is important to educate students and parents about the importance of eating breakfast, because it provides an important part of a student's daily intake of nutrients needed for energy, growth, and learning. PMID- 15801878 TI - Girls' experiences in physical education: competition, evaluation, & degradation. AB - School nurses are often asked to participate in the health component of many physical education (PE) programs in schools. With this opportunity comes an ability to invite a model of physical education that enables physical, mental, and relational health. A pilot study was initiated to explore why girls' enrollment in physical education was dwindling once the compulsory credits were achieved. What emerged was the reality that for many girls, physical education was a source of constant shaming regarding their athletic ability and eventually themselves. Forced competition, degrading evaluation, and sexuality- and size related harassment by both peers and teachers led the participants in this study to opt out of any further physical education classes. Within school-based physical education exists an opportunity to promote the health of adolescent women. School nurses, in partnership with physical education teachers and girls, can construct a model of physical education that is neither based on competition nor athletic ability, but rather on building a positive self-esteem and a positive relationship with one's body. This model would be rooted in participation, fun, and female friendship. PMID- 15801877 TI - Better Bones Buddies: an osteoporosis prevention program. AB - Although osteoporosis typically surfaces in later life, peak bone mass attained before age 20 is a key factor in its prevention. However, most American children's diets lack sufficient calcium during the critical growth periods of preadolescence and adolescence to achieve peak bone mass. Better Bones (BB) Buddies is an educational program targeting children ages 9-15 years in an effort to improve their knowledge of bone health and to increase their intake of calcium rich foods, thereby reducing the risk for osteoporosis later in life. In the 1998 1999 school year, Better Bones Buddies was given to more than 2,200 school children in southeastern South Dakota and southwestern Minnesota. Posttest results (N = 900) indicate participants improved in their knowledge of osteoporosis, and half reported modifications in their dietary habits to increase calcium consumption. Implications of the Better Bones Buddies program are discussed, with recommendations for future use of this program to increase children's knowledge about bone growth and osteoporosis. PMID- 15801879 TI - Virtual first impressions. AB - Frequently, a nurse's first and only contact with a graduate school, legislator, public health official, professional organization, or school nursing colleague is made through e-mail. The format, the content, and the appearance of the e-mail create a virtual first impression. Nurses can manage their image and the image of the profession by communicating electronically in a manner appropriate for the professional role. PMID- 15801881 TI - Lumps, bumps, and things that go itch in your office! PMID- 15801882 TI - Fecal incontinence, an ignored and concealed problem. PMID- 15801883 TI - Prevalence of silent fecal and urinary incontinence in women from the town of Teruel. AB - OBJECTIVES: To study the prevalence of fecal (FI) and urinary incontinence (UI) in women from Teruel (Spain), as well as the clinical conditions associated with these disorders. METHODS: We studied prospectively women with an age range of 20 64 years who were randomly selected from the population seen in a primary care center because of medical disorders not related to incontinence. Patients with functional or cognitive impairment were excluded. Medical and obstetric antecedents, as well as the type and frequency of incontinence symptoms were collected in a questionnaire. RESULTS: Out of 115 women, 103 completed the study (mean age: 41+/-12 years range 20-64). UI was present in 34.9% (stress 33%, urge 14%, mixed 47%), FI in 14 (13.6%) (flatus 57%, liquid stools 43%), and 10 (9.7%) displayed both disorders. Age > 42 years and body mass index more or equal of 25 were associated with FI and UI; pregnancy was only associated with UI, but the group of women with more or equal of 2 vaginal deliveries showed a higher frequency of FI (p < 0.05, Chi squared test). In the multivariate analysis, only the presence of UI was associated with FI (OR 6.0; CI 95% 1.7-21). Association of FI and UI was more frequent in women older than 42 years (OR 16.7, CI 95% 1.9 141). No statistical differences were found when smoking, exercise, and type of childbirth were compared between the presence/absence of FI or UI. CONCLUSIONS: Urinary and fecal incontinence are frequent in women, and the coexistence of both disorders is not uncommon. Age, overweight and parity are associated with the presence of fecal and/or urinary incontinence. PMID- 15801884 TI - Effect of ethanol consumption on colon cancer in an experimental model. AB - AIMS: The present study was designed to examine the effect of an ethanol supplement on experimental colon carcinogenesis. MATERIAL AND METHODS: One hundred and ten 10-week-old Sprague-Dawley rats were divided into five groups: group A (20 rats) received no treatment. Group B (20 rats) received a supplement of ethanol at 1.23 g/kg of body weight per day added to their drinking water for 24 weeks. Group C (30 rats) received 18 weekly doses of dimethylhydrazine (DMH) at 21 mg/kg of body weight from the beginning of the study. Group D (20 rats) received ethylen-diamin-tetracetic acid (EDTA) solution only for 18 weeks. Group E (20 rats) received ethanol at the same dose as group B plus DMH injections at the same dose as the rats in group C from the beginning of the study. All experimental animals were sacrificed after 25-27 weeks. RESULTS: No significant differences in the number of rats that developed tumors, number of tumor-free animals, and number of tumors per rat, as well as in macro-microscopic tumoral findings were observed for animals in group C compared to animals in group E. CONCLUSIONS: We concluded that the addition of an ethanol supplement does not modify colorectal carcinogenesis using a dynamic model of tumor induction with DMH. PMID- 15801885 TI - Doppler-guided hemorrhoidal artery ligation in the management of symptomatic hemorrhoids. AB - OBJECTIVE: The aim of this study is to clinically test the efficacy of a new approach for patients having symptomatic grade III and IV hemorrhoids. MATERIAL AND METHOD: 32 patients (17 females) complaining of grade III or IV hemorrhoids were included in the study. A specially designed proctoscope coupled with a Doppler transducer on its tip was used to identify the hemorrhoidal arteries, which were afterwards suture ligated. Operating time as well as per- and post operative complications were analyzed. Follow-up was planned following discharge after 1 week, 1 month, 6 months and 1 year. RESULTS: Mean operation time was 27 (range 18-43) minutes, and 5 (range 4-7) arteries were located on average. No patient had severe or moderate postoperative pain, with anal discomfort being the main complaint. Rectal bleeding and tenesmus were the commonest post-operative complications. After one year of follow-up, 19 patients were free of symptoms and 6 of them had significant symptom relief. According to grade, the technique failed in just 3 grade III patients, but in as many as 4 grade IV hemorrhoid cases. CONCLUSIONS: Doppler-guided hemorrhoid artery ligation is an easy-to perform technique that is well accepted by patients and has good results for grade III hemorrhoids. PMID- 15801886 TI - Mortality from colorectal cancer in Andalusia: findings in favor of mass screening. AB - OBJECTIVE: To describe mortality from colorectal cancer (CRC) in Andalusia from 1975 to 2001 in order to facilitate decision-making on mass screening. DESIGN: A population-based descriptive study. Considering mortality data from CRC and the resident population of Andalusia during the studied period, the main indicators of mortality by age and sex groups were calculated, and trends were estimated. RESULTS: There have been annual increases in mortality of 1.99% in men and 0.15% in women during the study period. In men, this is mainly due to increased mortality in those older than 65 years, and in women in those in the 35- to 64 year-old age group. In the younger age groups of both sexes, there has been a decline in mortality. The trend of mortality from colon cancer was positive in both men and women, but from rectal cancer was negative in women and slightly positive in men. CONCLUSIONS: Increased mortality from CRC in Andalusia may justify mass screening measures to potentially change mortality evolution, as has occurred in other countries. PMID- 15801887 TI - Hepatotoxicity associated with the ingestion of Centella asiatica. AB - BACKGROUND: Hepatotoxicity due to herbal remedies is being increasingly recognized. Centella asiatica (Centella asiatica Linn Urban) is commercialized for multiple conditions. Its active principles are pentacyclic triterpenic saponosides (asiaticoside, madecassoside). CLINICAL CASE STUDIES: We present three women (61, 52 and 49 years old) who developed jaundice after taking Centella asiatica for 30, 20 and 60 days. Respective laboratory tests: ALT: 1193, 1694 and 324 U/L; ALP: 503, 472 and 484 U/L; bilirubin: 4.23, 19.89 and 3.9 mg/dl. The first patient also had ASMA 1/160 and AMA 1/320. Respective pathological diagnoses: granulomatous hepatitis with marked necrosis and apoptosis; chronic hepatitis with cirrhotic transformation and intense necroinflammatory activity, and granulomatous hepatitis. All patients improved with Centella asiatica discontinuation, and ursodeoxycholic acid 10 mg/kg/day. The first patient took Centella asiatica again, with recurrence of the damage. The second one had taken this herb a year before. CONCLUSIONS: Many plants synthesize hepatotoxic compounds. Germander, Skullcap and Glycyrrhizin contain di or triterpenic active principles, which can produce hepatic injury by promoting apoptosis and altering cell membranes. We hypothesize that these mechanisms may have resulted in injuries associated with Centella asiatica. The presence of autoantibodies and granulomas also favors an immune-mediated mechanism. Ursodeoxycholic acid has anti-apoptotic properties, but we cannot rule out that Centella asiatica discontinuation alone may have resulted in patient improvement. PMID- 15801888 TI - Tridimensional (3D) ultrasonography. AB - A review and update on tridimensional ultrasonography covering all its aspects, ultrasonography and 3D endoscopic ultrasonography, its technical details and current indications, such as volume calculation, vascular architecture assessment, and role in interventionist ultrasounds. Images included are from the author's own experience. PMID- 15801889 TI - Bronchobiliary fistula secondary to biliary stricture after hepatectomy. PMID- 15801890 TI - [Gastrinoma of "normal" anatomic location]. PMID- 15801891 TI - [Endoscopic drainage of a liver hydatic cyst open to intrahepatic bile ducts]. PMID- 15801892 TI - Who should be responsible for sedation techniques in digestive endoscopy? PMID- 15801893 TI - DNA aneuploidy in colorectal adenomas. Role in the adenoma-carcinoma sequence. AB - INTRODUCTION: Aneuploidy has been observed in 6-27% of lesions known to be precursors of colorectal cancer, such as adenomas or ulcerative colitis. It has been suggested that aneuploidy may predispose to malignancy in these cases. However, its role in the adenoma-carcinoma sequence has not been definitely established. The objective of this study was to assess the incidence of aneuploidy in colon adenomas, as well as to study its possible role in the adenoma-carcinoma sequence. MATERIAL AND METHODS: The study was performed on a series of 57 large bowel adenomas measuring 10 mm or more, collected from 54 consecutive patients. All specimens were obtained either by endoscopic or by surgical resection. There were 49 adenomas with low-grade dysplasia, two with high-grade dysplasia, two intramucous carcinomas, and four microinvasive carcinomas. A flow cytometric DNA analysis was performed in fresh specimens following Vindelov's method. RESULTS: Aneuploid DNA was detected in five out of 49 low-grade dysplasia adenomas (10%), in all four high-grade dysplasia adenomas or intramucous carcinomas (100%), and in three out of four microinvasive carcinomas (75%). The association between aneuploidy and high-grade dysplasia adenomas, intramucous, or microinvasive carcinoma was statistically significant (p < 0.001). No association was found between aneuploidy and any of the following features: age, gender, clinical symptoms of patients, and size or location of adenomas. CONCLUSIONS: The incidence of aneuploidy in this series was 10% in low grade dysplasia adenomas, and 87% in high-grade dysplasia adenomas or carcinomas, and this difference was statistically significant. These findings suggest that aneuploidy may play a role in the adenoma-carcinoma sequence. PMID- 15801894 TI - A pilot study on the endoscopic surveillance of colorectal dysplasia and cancer in long-standing ulcerative colitis. AB - INTRODUCTION: Patients with ulcerative colitis (UC) have a greater risk of developing colorectal cancer (CRC) when compared to the general population. Epithelial dysplasia comes before this neoplasm, and thus endoscopic surveillance is recommended to these patients. This pilot study aims at establishing the incidence of dysplasia and CRC in patients with long-standing UC in our hospital. MATERIAL AND METHODS: This is a prospective observational study performed in patients with a definite diagnosis of UC for more than 8 years. These patients were encouraged to enroll in an endoscopic surveillance program for CRC. All patients underwent colonoscopy and multiple biopsies every 18 to 24 months in order to detect epithelial dysplasia. RESULTS: Thirty-nine patients were included from January 1994 to December 2003. Half of them were males. Mean age was 52 +/- 13 years. Mean duration of UC was 15 +/- 8 years. Thirteen (35%) patients had left colitis, and 26 (65%) had pancolitis or extensive colitis. The presence of mild dysplasia was detected in four patients, on two occasions in one of them (13%; 95% CI: 6.1-33.5); the incidence of mild dysplasia was 1.3% patients per surveillance year. No severe dysplasia or CRCs were identified. CONCLUSION: The incidence of dysplasia in our area is lower than expected, and does not support surveillance programs for these patients. However, no definite conclusions may be drawn from such a small number of patients. PMID- 15801895 TI - Sedation at endoscopic units in Galicia: results of the "Sociedad Gallega de Patologia Digestiva" inquiry. AB - AIM: To evaluate the human and material resources available for sedation, and the usual manner of handling them at endoscopic units in Galicia. METHODS: A prospective and descriptive study based on the performance, distribution, and analysis of a clinical practice inquiry. We requested information about endoscopies performed, available means for sedation, sedation monitoring, and level of sedation used in each procedure. RESULTS: Our inquiry was answered by twenty endoscopic units (thirteen were in public hospitals, and eleven performed complex procedures). Of these units, 80% had a pulse oximeter, 42% had continuous electrocardiography, 40% had a defibrillator, and 45% had a recovery area. The drug most commonly used in gastroscopies was midazolam (76%), and the combination midazolam-meperidine was most frequent in both colonoscopies (72%) and ERCPs (60%). An anesthesiologist was usually available for certain procedures in 15% of units, and as an exception in 65%. Of those inquired, 35% wished to have a full time anesthesiologist in the unit, 25% wished to have an anesthetist only for certain procedures, and 35% on an exceptional basis. Finally, endoscopists considered that 83% of therapeutical gastroscopies, 87% of therapeutical colonoscopies, 98% of ERCPs, 95% of enteroscopies, and 98% of echoendoscopies deserved sedation. CONCLUSIONS: Although endoscopists consider that endoscopic procedures should benefit from sedation in a high proportion, the available resources to safely monitor patients are inadequate in some units. PMID- 15801896 TI - Rubber-band ligation for hemorrhoids in a colorectal unit. A prospective study. AB - INTRODUCTION: Nowadays the rubber band ligation technique is one of the most worldwide used and effective treatment of the hemorrhoidal disease. OBJECTIVES: Our study has as a goal to analyze the success or failure of the rubber band ligation in hemorrhoids grade 1,2 and 3, to analyze their complications and to see if all symptomatic hemorrhoids should be treated with ligation at the first visit. PATIENTS AND METHOD: A prospective and descriptive study was designed for patients who came to the Colorectal Unit with hemorrhoidal disease from September 1997 to December 2001. First, second and third degree patients were treated according to the classification of hemorrhoids of St. Mark's Hospital. The technique of ligation after Barron was applied. RESULTS: From 261 patients with a mean age of 48.3 (range: 16-86), 181 (99 M/82 W) have been treated with Barron's method and 80 with rich fiber diet and water. Rectal bleeding was the most common symptom (91.16%). Anuscopy showed hemorrhoidal disease in all the cases. From 181 patients, 19.33% were hemorrhoids degree I, 51.93% degree II and 29.83% degree III. Two hundred and eighty-seven ligation sessions were done and the balance of ligations per patient was 2.45 and 1.5 rubber band per session. The 32% of the patients referred pain after ligation. A 13.81% of cases were operated due to persistent rectal bleeding or hemorrhoidal prolapse. CONCLUSIONS: Symptomatic hemorrhoids degree I and II with a short clinical history should be treated initially with a rich fiber and water diet. The technique of Barron is an effective therapy to treat the hemorrhoids degree 1, 2 and in 74% of success in cases with degree 3. PMID- 15801897 TI - Diagnostic and treatment recommendations on perianal Crohn's disease. AB - Treatment of perianal fistulas in Crohn's disease should be defined on an individual basis. A combined medical and surgical approach is the optimal treatment. Adequate management of perianal fistula disease is based on the presence or absence of active proctitis, anatomic location, and fistula type. Furthermore, the presence of perianal abscesses must be ruled out. This evaluation includes digital rectal examination, endoscopy, and examination under anesthesia combined with pelvic magnetic resonance imaging or anorectal endoscopy ultrasonography findings. PMID- 15801898 TI - Severe gastrointestinal bleeding associated to massive jejunal diverticulosis and Sintrom treatment. PMID- 15801899 TI - [Ileal perforation secondary to cytomegalovirus infection]. PMID- 15801900 TI - [Lung adenocarcinoma diagnosed by the presence of duodenal and jejunal metastases]. PMID- 15801901 TI - [Sporadic gastric polyposis]. PMID- 15801902 TI - [Splenic cysts: clinical, diagnostic and therapeutic aspects]. PMID- 15801903 TI - [Colic ischemia mimicking carcinoma]. PMID- 15801904 TI - Cardiovascular risk parameters in men with ankylosing spondylitis in comparison with non-inflammatory control subjects: relevance of systemic inflammation. AB - Men with AS (ankylosing spondylitis) are at elevated risk for CHD (coronary heart disease) but information on risk factors is sparse. We compared a range of conventional and novel risk factors in men with AS in comparison with healthy controls and, in particular, determined the influence of systemic inflammation. Twenty-seven men with confirmed AS and 19 controls matched for age were recruited. None of the men was taking lipid-lowering therapy. Risk factors inclusive of plasma lipids, IL-6 (interleukin-6), CRP (C-reactive protein), vWF (von Willebrand factor), fibrin D-dimer, ICAM-1 (intercellular cell-adhesion molecule-1) and fibrinogen were measured, and blood pressure and BMI (body mass index) were determined by standard techniques. A high proportion (70%) of men with AS were smokers compared with 37% of controls (P = 0.024). The AS patients also had a higher BMI. In analyses adjusted for BMI and smoking, men with AS had significantly higher IL-6 and CRP (approx. 9- and 6-fold elevated respectively; P < 0.001), fibrinogen (P = 0.013) and vWF (P = 0.008). Total cholesterol and HDL-C (high-density lipoprotein cholesterol) were lower (P < 0.05 and P = 0.073 respectively) in AS and thus the ratio was not different. Pulse pressure was also significantly higher in AS (P = 0.007). Notably, adjustment for IL-6 and CRP levels rendered all case-control risk factor differences, except pulse pressure, non-significant. In accordance with this finding, IL-6 correlated positively (r = 0.74, P < 0.001) with fibrinogen, but negatively (r = -0.46, P = 0.016) with total cholesterol concentration. In conclusion, men with AS have perturbances in several CHD risk factors, which appear to be driven principally by systemic inflammatory mediators. Inflammation-driven atherogenesis potentially contributes to the excess CHD risk in AS. PMID- 15801905 TI - Novel variants of human SCaMC-3, an isoform of the ATP-Mg/P(i) mitochondrial carrier, generated by alternative splicing from 3'-flanking transposable elements. AB - CaMCs (calcium-dependent mitochondrial carriers) represent a novel subfamily of metabolite carriers of mitochondria. The ATP-Mg/P(i) co-transporter, functionally characterized more than 20 years ago, has been identified to be a CaMC member. There are three isoforms of the ATP-Mg/P(i) carrier in mammals, SCaMC-1 (short CaMC-1), -2 and -3 (or APC-1, -3 and -2 respectively), corresponding to the genes SLC25A24, SLC25A25 and SLC25A23 respectively, as well as six N-terminal variants generated by alternative splicing for SCaMC-1 and -2 isoforms. In the present study, we describe four new variants of human SCaMC-3 generated by alternative splicing. The new mRNAs use the exon 9 3'-donor site and distinct 5'-acceptor sites from repetitive elements, in regions downstream of exon 10, the last exon in all SCaMCs. Transcripts lacking exon 10 (SCaMC-3b, -3b', -3c and -3d) code for shortened proteins lacking the last transmembrane domain of 422, 456 and 435 amino acids, and were found in human tissues and HEK-293T cells. Mitochondrial targeting of overexpressed SCaMC-3 variants is incomplete. Surprisingly, the import impairment is overcome by removing the N-terminal extension of these proteins, suggesting that the hydrophilic N-terminal domain also participates in the mitochondrial import process, as shown for the CaMC members aralar and citrin [Roesch, Hynds, Varga, Tranebjaerg and Koehler (2004) Hum. Mol. Genet. 13, 2101 2111]. PMID- 15801906 TI - Proteomic detection of hydrogen peroxide-sensitive thiol proteins in Jurkat cells. AB - Thiol proteins are important in cellular antioxidant defenses and redox signalling. It is postulated that reactive oxidants cause selective thiol oxidation, but relative sensitivities of different cell proteins and critical targets are not well characterized. We exposed Jurkat cells to H2O2 for 10 min and measured changes in reversibly oxidized proteins by labelling with iodoacetamidofluorescein and two-dimensional electrophoresis. At 200 microM H2O2, which caused activation of the MAP (mitogen-activated protein) kinase ERK (extracellular-signal-regulated kinase), growth arrest and apoptosis, relatively few changes were seen. A total of 28 spots were reversibly oxidized (increased labelling intensity) and 24 decreased. The latter included isoforms of peroxiredoxins 1 and 2, which were irreversibly oxidized. Oxidation of GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was striking, and other affected proteins included glutathione S-transferase P1-1, enolase, a regulatory subunit of protein kinase A, annexin VI, the mitotic checkpoint serine/threonine-protein kinase BUB1beta, HSP90beta (heat-shock protein 90beta) and proteosome components. At 20 microM H2O2, changes were fewer, but GAPDH and peroxiredoxin 2 were still modified. Dinitrochlorobenzene treatment, which inhibited cellular thioredoxin reductase and partially depleted GSH, caused reversible oxidation of several proteins, including thioredoxin 1 and peroxiredoxins 1 and 2. Most changes were distinct from those with H2O2, and changes with H2O2 were scarcely enhanced by dinitrochlorobenzene. Relatively few proteins, including deoxycytidine kinase, nucleoside diphosphate kinase and a proteosome activator subunit, responded only to the combined treatment. Thus most of the effects of H2O2 were not linked to thioredoxin oxidation. Our study has identified peroxiredoxin 2 and GAPDH as two of the most oxidant-sensitive cell proteins and has highlighted how readily peroxiredoxins undergo irreversible oxidation. PMID- 15801908 TI - IGF-I stimulation of proteoglycan synthesis by chondrocytes requires activation of the PI 3-kinase pathway but not ERK MAPK. AB - The IGF-I (insulin-like growth factor-I) signalling pathway responsible for regulation of proteoglycan synthesis in chondrocytes has not been defined and is the focus of the present study. Chondrocytes isolated from normal human articular cartilage were stimulated with IGF-I in monolayer culture or in suspension in alginate. IGF-I activated members of both the PI3K (phosphoinositide 3-kinase) pathway and the ERK (extracellular-signal-regulated kinase)/MAPK (mitogen activated protein kinase) pathway. The PI3K inhibitors LY294002 and wortmannin blocked IGF-I-stimulated Akt phosphorylation without blocking ERK phosphorylation and this was associated with complete inhibition of proteoglycan synthesis. A decrease in IGF-I-stimulated proteoglycan synthesis was also observed upon inhibition of mTOR (mammalian target of rapamycin) and p70S6 kinase, both of which are downstream of Akt. The MEK (MAPK/ERK kinase) inhibitors PD98059 and U0126 blocked IGF-I-stimulated ERK phosphorylation but did not block the phosphorylation of Akt and did not decrease proteoglycan synthesis. Instead, in alginate- cultured chondrocytes, the MEK inhibitors increased IGF-I-stimulated proteoglycan synthesis when compared with cells treated with IGF-I alone. This is the first study to demonstrate that IGF-I stimulation of the PI3K signalling pathway is responsible for the ability of IGF-I to increase proteoglycan synthesis. Although IGF-I also activates the ERK/MAPK pathway, ERK activity is not required for proteoglycan synthesis and may serve as a negative regulator. PMID- 15801907 TI - Sarcolipin and phospholamban mRNA and protein expression in cardiac and skeletal muscle of different species. AB - The widely held view that SLN (sarcolipin) would be the natural inhibitor of SERCA1 (sarcoplasmic/endoplasmic-reticulum Ca2+-ATPase 1), and PLB (phospholamban) its counterpart for SERCA2 inhibition is oversimplified and partially wrong. The expression of SLN and PLB mRNA and protein relative to SERCA1 or SERCA2 was assessed in ventricle, atrium, soleus and EDL (extensor digitorum longus) of mouse, rat, rabbit and pig. SLN protein levels were quantified by means of Western blotting using what appears to be the first successfully generated antibody directed against SLN. Our data confirm the co expression of PLB and SERCA2a in cardiac muscle and the very low levels (in pig and rabbit) or the absence (in rat and mouse) of PLB protein in the slow skeletal muscle. In larger animals, the SLN mRNA and protein expression in the soleus and EDL correlates with SERCA1a expression, but, in rodents, SLN mRNA and protein show the highest abundance in the atria, which are devoid of SERCA1. In the rodent atria, SLN could therefore potentially interact with PLB and SERCA2a. No SLN was found in the ventricles of the different species studied, and there was no compensatory SLN up-regulation for the loss of PLB in PLB(-/-) mouse. In addition, we found that SLN expression was down-regulated at the mRNA and protein level in the atria of hypertrophic hearts of SERCA2(b/b) mice. These data suggest that superinhibition of SERCA by PLB-SLN complexes could occur in the atria of the smaller rodents, but not in those of larger animals. PMID- 15801909 TI - Specific effect of arachidonic acid on inducible nitric oxide synthase mRNA expression in human osteoblastic cells. AB - A specific modulatory effect of PUFAs (polyunsaturated fatty acids) on gene expression of some cytokines involved in bone remodelling has been reported previously. In particular, although a direct action of AA (arachidonic acid) on bone cytokine gene expression has been shown in human osteoblastic cells, OA (oleic acid) and EPA (eicosapentaenoic acid) were ineffective. Since the NO (nitric oxide) system has also been shown to have an important modulatory activity on osteoblasts, osteoclasts and bone metabolism, in the present study we have investigated the effects of PUFAs on iNOS (inducible NO synthase) gene expression in a human osteoblast-like cell line. AA induced a significant increase in iNOS mRNA expression, whereas EPA and OA had no stimulatory effects but instead caused a significant inhibition of AA-induced iNOS gene expression. Blocking of the COX (cyclo-oxygenase) pathway did not inhibit AA-induced iNOS expression. AA action was inhibited instead by the addition of calphostin C and genistein, inhibitors of PKC (protein kinase C) and tyrosine kinases respectively. Experiments performed with specific anti-cytokine antibodies showed a significant decrease in iNOS expression in AA-treated osteoblastic cells, suggesting that both cytokine-dependent and -independent mechanisms account for the effects of AA on iNOS gene expression. In conclusion, our investigation clearly shows specific effects of PUFAs on iNOS expression in human osteoblast like cells with a cytokine-dependent and -independent mechanism. These results might have clinical relevance and are of interest for understanding the reported beneficial effects of dietary PUFA manipulation on the prevention and/or treatment of primary and secondary bone disease. PMID- 15801910 TI - Argatroban-coupled Affi-Gel matrix for the purification of thrombin from plasma. AB - Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay. None of the available purification methods easily deals with this subject. The procedure described in the present paper uses a readily available pharmaceutical agent, argatroban, to construct an affinity matrix. Argatroban has a high affinity for thrombin and its thrombin binding is reversible. Prothrombin derived from a Ba(2+) precipitate of human plasma is used as the starting material. The crude prothrombin can be bulk activated to thrombin using taipan-snake (Oxyuranus scutellatus) venom and bound to the argatroban coupled matrix without further processing steps. The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis. This purification scheme is rapid, yielding purified thrombin within 2 days. PMID- 15801911 TI - The histidine of the c-type cytochrome CXXCH haem-binding motif is essential for haem attachment by the Escherichia coli cytochrome c maturation (Ccm) apparatus. AB - c-type cytochromes are characterized by covalent attachment of haem to the protein by two thioether bonds formed between the haem vinyl groups and the cysteine sulphurs in a CXXCH peptide motif. In Escherichia coli and many other Gram-negative bacteria, this post-translational haem attachment is catalysed by the Ccm (cytochrome c maturation) system. The features of the apocytochrome substrate required and recognized by the Ccm apparatus are uncertain. In the present study, we report investigations of maturation of cytochrome b562 variants containing CXXCR, CXXCK or CXXCM haem-binding motifs. None of them showed any evidence for correct maturation by the Ccm system. However, we have determined, for each variant, that the proteins (i) were expressed in large amounts, (ii) could bind haem in vivo and/or in vitro and (iii) were not degraded in the cell. Together with previous observations, these results strongly suggest that the apocytochrome substrate feature recognized by the Ccm system is simply the two cysteine residues and the histidine of the CXXCH haem-binding motif. Using the same experimental approach, we have also investigated a cytochrome b562 variant containing the special CWSCK motif that binds the active-site haem of E. coli nitrite reductase NrfA. Whereas a CWSCH analogue was matured by the Ccm apparatus in large amounts, the CWSCK form was not detectably matured either by the Ccm system or by the dedicated Nrf biogenesis proteins, implying that the substrate recognition features for haem attachment in NrfA may be more extensive than the CWSCK motif. PMID- 15801913 TI - Identification of trypanothione from the human pathogen Entamoeba histolytica by mass spectrometry and chemical analysis. AB - In this paper, we present definitive data to show, from ESI (electrospray ionization) studies, that the thiol-bimane compound isolated and purified from Entamoeba histolytica trophozoites, corresponds unequivocally to the structure of trypanothione. Trypanothione disulphide was shown to have a molecular ion of m/z 722. It was further demonstrated by MALDI-TOF (matrix-assisted laser desorption ionization-time-of-flight) MS that this thiol compound also corresponds to the characteristic monoprotonated ion of trypanothione-(bimane)(2), which has a molecular ion of m/z 1103.95. The ion pattern of the thiol-bimane compound prepared from the commercial trypanothione standard is identical with the E. histolytica thiol-bimane compound. After HPLC separation, chemical amino acid analysis by dabsylation and dansylation of the thiol bimane compound from Entamoeba showed the presence of the following trypanothione components: glutamic acid, cysteic acid, glycine and spermidine. We can conclude from these highly reliable MS experiments and chemical analyses that E. histolytica contains the thiol compound trypanothione, which was previously thought to occur only in trypanosomatids. PMID- 15801914 TI - Systematic review and meta-analysis: is 1-week proton pump inhibitor-based triple therapy sufficient to heal peptic ulcer? AB - AIMS: To systematically review the efficacy on ulcer healing of 1-week combination of a proton pump inhibitor plus two antibiotics and to perform a meta analysis of randomized clinical trials to evaluate whether 7 days of proton pump inhibitor-based triple therapy is sufficient to heal peptic ulcer. METHODS: Studies where 1-week proton pump inhibitor-based triple therapy was administered to heal peptic ulcer were included. Randomized clinical trials comparing the efficacy on ulcer healing of 7-day proton pump inhibitor-based triple therapy versus this same regimen but prolonging the proton pump inhibitor for several more weeks were included in the meta-analysis. Electronic and manual bibliographical searches were conducted. Meta-analysis was performed combining the odds ratios of the individual studies. RESULTS: Twenty-four studies (2342 patients) assessed ulcer healing with 1-week proton pump inhibitor-based triple therapy. Mean healing rate was 86%, and 95% in Helicobacter pylori-eradicated patients. Six studies (862 patients), were included in the meta-analysis. Mean ulcer healing rate with a 7-day treatment was 91% versus 92% when proton pump inhibitor was prolonged for 2-4 more weeks (odds ratio = 1.11; 95% confidence interval = 0.71-1.74). CONCLUSION: In patients with peptic ulcer and H. pylori infection, prolonging therapy with proton pump inhibitor after a triple therapy for 7 days with a proton pump inhibitor and two antibiotics is not necessary to induce ulcer healing. PMID- 15801912 TI - Integral membrane lipid phosphatases/phosphotransferases: common structure and diverse functions. AB - Phospholipids and sphingolipids play critical roles in signal transduction, intracellular membrane trafficking, and control of cell growth and survival. We discuss recent progress in the identification and characterization of a family of integral membrane proteins with central roles in bioactive lipid metabolism and signalling. These five groups of homologous proteins, which we collectively term LPTs (lipid phosphatases/phosphotransferases), are characterized by a core domain containing six transmembrane-spanning alpha-helices connected by extramembrane loops, two of which interact to form the catalytic site. LPT family members are localized to all major membrane compartments of the cell. The transmembrane topology of these proteins places their active site facing the lumen of endomembrane compartments or the extracellular face of the plasma membrane. Sequence conservation between the active site of the LPPs (lipid phosphate phosphatases), SPPs (sphingosine phosphate phosphatases) and the recently identified SMSs (sphingomyelin synthases) with vanadium-dependent fungal oxidases provides a framework for understanding their common catalytic mechanism. LPPs hydrolyse LPA (lysophosphatidic acid), S1P (sphingosine 1-phosphate) and structurally-related substrates. Although LPPs can dephosphorylate intracellularly generated substrates to control intracellular lipid metabolism and signalling, their best understood function is to regulate cell surface receptor-mediated signalling by LPA and S1P by inactivating these lipids at the plasma membrane or in the extracellular space. SPPs are intracellularly localized S1P-selective phosphatases, with key roles in the pathways of sphingolipid metabolism linked to control of cell growth and survival. The SMS enzymes catalyse the interconversion of phosphatidylcholine and ceramide with sphingomyelin and diacylglycerol, suggesting a pivotal role in both housekeeping lipid synthesis and regulation of bioactive lipid mediators. The remaining members of the LPT family, the LPR/PRGs (lipid phosphatase-related proteins/plasticity-related genes) and CSS2s (type 2 candidate sphingomyelin synthases), are presently much less well studied. These two groups include proteins that lack critical amino acids within the catalytic site, and could therefore not use the conserved LPT reaction mechanism to catalyse lipid phosphatase or phosphotransferase reactions. In this review, we discuss recent ideas about their possible biological activities and functions, which appear to involve regulation of cellular morphology and, possibly, lipid metabolism and signalling in the nuclear envelope. PMID- 15801915 TI - Long-term treatment of gastro-oesophageal reflux disease patients with frequent symptomatic relapses using rabeprazole: on-demand treatment compared with continuous treatment. AB - BACKGROUND: On-demand treatment may be an alternative in the long-term treatment of non-severe gastro-oesophageal reflux disease in patients with frequent symptomatic relapses. AIM: To compare the efficacy of on-demand treatment with rabeprazole 10 mg versus continuous treatment in the long-term treatment of patients with frequent symptomatic relapses of mild to moderate gastro oesophageal reflux disease. METHODS: This randomized, open-label study enrolled patients diagnosed with non-erosive reflux disease or oesophagitis grade 1 or 2 (Savary-Miller classification) reporting frequent symptomatic relapses (requiring > or =2 courses of antisecretory therapy during the previous year), whose intensity is rated at least moderate (>2 on a 5-point Likert scale). After a 4 week selection phase with rabeprazole 10 mg once daily, patients reporting symptom relief (Likert score < or =2) were randomized to receive either rabeprazole 10 mg continuous treatment or on-demand treatment for 6 months. The main evaluation criterion was the rate of symptom relief (scored on the Likert scale) after 6 months. RESULTS: One hundred and seventy-six patients were enrolled in the 4-week selection phase (men, 53%; mean age, 49 years; non-erosive reflux disease, 36.4%; gastro-oesophageal reflux disease 1, 53.4%; gastro oesophageal reflux disease 2, 10.2%). Rabeprazole relieved symptoms in 88.6% of patients. Of this group, 152 were randomized to the comparative phase to receive rabeprazole 10 mg continuous treatment (once daily) or on-demand treatment (continuous treatment, n = 81; on-demand treatment, n = 71). At month 6 (end point), the symptom relief rate was slightly higher for patients in the continuous treatment group compared with those in the on-demand treatment group: 86.4% versus 74.6%, respectively. This difference was not statistically significant (P = 0.065). For the overall quality of life score, there was no difference between the continuous treatment and on-demand treatment groups (86.25 and 84.94). Mean daily consumption of rabeprazole was significantly lower in the on-demand treatment group versus the continuous treatment group (0.31 tablets versus 0.96 tablets; P < 0.0001). CONCLUSION: On-demand therapy with rabeprazole 10 mg provides an alternative to continuous therapy in patients with mild to moderate gastro-oesophageal reflux disease suffering from frequent symptomatic relapses. PMID- 15801916 TI - Missing gastric cancer in dyspepsia. AB - BACKGROUND: Dyspepsia is common in gastric cancer, but also in many benign conditions. European Helicobacter pylori Study Group and American Gastroenterological Association guidelines recommend endoscopy in dyspepsia for patients with alarm symptoms or at age >45 years. However, recommendations are controversial. AIM: To investigate whether criteria for endoscopy in patients with dyspepsia are adequate to detect gastric cancer. METHODS: In 215 patients at initial diagnosis of gastric adenocarcinoma, symptoms were classified as alarm and non-alarm. Cases were staged according to the TNM system. Stages T(1) T(3)N(x)M(0) were defined as potentially curable. RESULTS: Dyspepsia was present in 128 patients. Among patients with dyspepsia, 15 were < or =45 years and 41 denied alarm symptoms. The combination of both criteria excluded only three (2.3%) patients from endoscopy, but increasing the threshold to >50 and >55 years would have raised the rate of excluded patients to seven (5.5%) and 11 (8.6%). Only 53 potentially curable stages and 18 early gastric cancers occurred, but the tumour stage was not associated with dyspepsia duration, age threshold of 45 years, or alarm symptoms. CONCLUSIONS: Our results support current European Helicobacter Study Group and American Gastroenterological Association criteria for endoscopy in patients with dyspepsia to detect gastric cancer. Regardless, most cancers are advanced at detection. PMID- 15801917 TI - Risk factors for gastro-oesophageal reflux disease symptoms: a community study. AB - AIM: To examine the prevalence of gastro-oesophageal reflux disease symptoms and potential risk factors among community subjects. METHODS: A questionnaire was sent to 4000 subjects, stratified by age, gender and ethnicity to be representative of the local population. Gastro-oesophageal reflux disease symptoms were defined as at least weekly heartburn or acid regurgitation. RESULTS: 2231 responded (59%), 691 refused to participate and seven were incomplete. 1533 (41%) were evaluable (637 male, mean age 51 years, range: 20 80). The prevalence of gastro-oesophageal reflux disease symptoms was 21%. Smoking, excess alcohol, irritable bowel syndrome, increasing body mass index, a family history of upper gastrointestinal disease, increasing Townsend deprivation index, anticholinergic drugs (all P < 0.0001), weight gain, antidepressant drugs, inhaled bronchodilators, no educational attainment (all P < 0.01), south Asian origin (P = 0.02) and manual work (P < 0.05) were associated with gastro oesophageal reflux disease symptoms. Multivariate logistic regression revealed increasing body mass index, a family history of upper gastrointestinal disease, irritable bowel syndrome, south Asian origin (all P < 0.0001), smoking, excess alcohol, no educational attainment and anticholinergic drugs (all P < 0.01) were independently associated with gastro-oesophageal reflux disease symptoms. CONCLUSIONS: Frequent gastro-oesophageal reflux disease symptoms affect 21% of the population. Increasing body mass index, a family history of upper gastrointestinal disease, irritable bowel syndrome, south Asian origin, smoking, excess alcohol, social deprivation and anticholinergic drugs are independently associated with gastro-oesophageal reflux disease symptoms. PMID- 15801918 TI - 6-tioguanine monitoring in steroid-dependent patients with inflammatory bowel diseases receiving azathioprine. AB - BACKGROUND: 6-Thioguanine (6-tioguanine) nucleotides are the active metabolites of azathioprine. AIM: The aim of the study was to evaluate the rate of clinical remission without steroids in steroid-dependent Crohn's disease and ulcerative colitis patients receiving azathioprine, the medium- and long-term efficacy and the predictive factors of clinical response when monitoring 6-tioguanine. METHODS: Steroid-dependent Crohn's disease and ulcerative colitis patients receiving either azathioprine or not (treated later with a daily dose of 2.5 mg/kg) were prospectively included. 6-tioguanine was monitored at 1 and 2 months and every 3 months thereafter for 1 year. The azathioprine dose was adapted to reach a 6-tioguanine level of >250 pmol/8 x 10(8) red blood cells. Thiopurine methyltransferase genotype/phenotype was evaluated in some patients. RESULTS: A total of 106 patients were prospectively included (70 Crohn's disease, 36 ulcerative colitis). The clinical remission rate without steroids in patients receiving azathioprine, in intention-to-treat analysis, was 72% and 59% at 6 and 12 months, respectively. The remission rate was significantly higher in patients with 6-tioguanine >250 pmol/8 x 10(8) RBC (86% and 69% at 6 and 12 months, respectively; P < 0.01). No significant difference was observed between Crohn's disease and ulcerative colitis patients whether treated by azathioprine or not on inclusion. In the univariate analysis, the absence of Crohn's disease stenosis, a 6-tioguanine level >250 pmol/8 x 10(8) RBC, and an increase of erythrocyte mean corpuscular volume were the factors predictive of a favourable clinical response. In the multivariate analysis, only a 6-tioguanine level of >250 pmol/8 x 10(8) red blood cells was a predictive factor of favourable clinical remission. CONCLUSIONS: Clinical remission without steroids is significantly more likely when monitoring 6-tioguanine so as to reach a level of >250 pmol/8 x 10(8) red blood cells in steroid-dependent Crohn's disease and ulcerative colitis patients receiving azathioprine (86% and 69% at 6 and 12 months, respectively). PMID- 15801919 TI - Liver histology of Asian patients with chronic hepatitis B on prolonged lamivudine therapy. AB - BACKGROUND: Long-term effect of YMDD mutations on liver histology in Chinese hepatitis B patients is unknown. AIM: To examine the effect of prolonged lamivudine treatment on liver histology in Chinese patients with and without YMDD mutations. METHODS: Liver histology was assessed in 85 patients on long-term lamivudine at baseline and year 1, and at year 3 for 25 patients. RESULTS: Comparing patients with and without YMDD mutations at year 1, the former had higher baseline median necroinflammatory (11 vs. six respectively, P = 0.014) and fibrosis scores (three vs. one respectively, P = 0.001). The proportion of patients with improvement in necroinflammation and worsening of fibrosis was comparable for patients with and without YMDD mutations at year 1 (57.1%, 14.3% vs. 55%, 15% respectively) and year 3 (57.9%, 26.3% vs. 50%, 16.7% respectively). Comparing the histology at year 1 and 3, more patients with YMDD mutations developing after year 1 had worsening of necroinflammation than patients with persistent YMDD wild type (53.8% vs. 25% respectively). CONCLUSIONS: Patients who developed YMDD mutations had higher baseline histological scores. With YMDD mutations, the liver histology became less favourable after 3 years than at the first year, although there was still improvement when compared with that at baseline. PMID- 15801920 TI - Validity of 13C-phenylalanine breath test to evaluate functional capacity of hepatocyte in patients with liver cirrhosis and acute hepatitis. AB - BACKGROUND: No definitive method for quantitative evaluation of hepatic function has as yet been established. AIM: To investigate whether the (13)C-phenylalanine breath test would be useful for the evaluation of hepatic function in patients with liver cirrhosis and acute hepatitis. METHODS: L-[1-(13)C]-phenylalanine was administered orally in a dose of 100 mg to 25 patients with liver cirrhosis, 22 patients with acute hepatitis and 10 healthy subjects. The relationships of the cumulative excretion with the (13)C-%dose/h, blood biochemical parameters and asialoscintigraphy were investigated. RESULTS: In liver cirrhosis patients, the cumulative excretion showed correlations with hepatic function tests, asialoscintigraphy, clinical stage and portal hypertension. In acute hepatitis patients, the cumulative excretion showed correlations with hepatic function tests. There were positive correlations between the cumulative excretion and the (13)C-%dose/h at 20 min (Phe20) and (13)C-%dose/h at 30 min (Phe30) in liver cirrhosis and acute hepatitis patients. Multiple regression analysis demonstrated that total bilirubin, total cholesterol and absence of varices were independent determinants of cumulative excretion in liver cirrhosis patients and prothrombin time in acute hepatitis patients. CONCLUSION: The (13)C-phenylalanine breath test may allow hepatic function to be evaluated non-invasively in liver cirrhosis and acute hepatitis patients, and the Phe20 and Phe30 may be useful for determination of function at a single time-point. PMID- 15801921 TI - De novo HCV infection among dialysis patients: a prospective study by HCV core antigen ELISA assay. AB - BACKGROUND: Dialysis patients remain a high-risk group for hepatitis C virus infection. The current diagnosis of hepatitis C virus in dialysis patients includes serological measurement of anti-hepatitis C virus antibody; however, nucleic acid amplification technology for assessing hepatitis C virus viraemia is commonly used in other populations. An enzyme-linked immunosorbent assay test for detecting antibody to hepatitis C nucleocapsid core antigen (hepatitis C virus core antigen) in human serum has been recently developed (hepatitis C virus Core Antigen enzyme-linked immunosorbent assay test). It is conceived for screening of donor blood products to significantly reduce the 'serologic window' occurring before seroconversion during acute hepatitis C virus. AIM AND METHODS: A cohort (n = 72) of patients on maintenance haemodialysis in a single unit in the years 2000-2003 was included. Study patients were tested monthly by hepatitis C virus Core Antigen enzyme-linked immunosorbent assay in a prospective, clinical trial. Routine results obtained by hepatitis C virus Core Antigen enzyme-linked immunosorbent assay test were confirmed by assessing hepatitis C virus viraemia by branched-chain DNA (bDNA) signal amplification assay. RESULTS: De novo hepatitis C virus infection was identified in three patients during the study period; the hepatitis C virus incidence was 1.38% (95% confidence intervals, 1.31 4.09) per year. In each patient, hepatitis C virus core antigen testing allowed the serological identification of acute hepatitis C virus before anti-hepatitis C virus seroconversion. Hepatitis C virus RNA testing confirmed the results obtained by hepatitis C virus Core Antigen enzyme-linked immunosorbent assay in all cases. The time from initial hepatitis C virus detection by hepatitis C virus Core Antigen Assay and anti-hepatitis C virus seroconversion was not greater than four weeks. Two (67%) of three patients with de novo hepatitis C virus acquisition were HBsAg negative; both these patients underwent an initial phase of hepatitis C virus viraemia that was associated with an increase in alanine aminotransferase activity and preceded the seroconversion to anti-hepatitis C virus antibody. Nosocomial transmission of hepatitis C virus between haemodialysis patients was implicated in at least two (67%) of these three patients. CONCLUSIONS: Serological testing for hepatitis C virus core antigen can identify acute hepatitis C virus infection before anti-hepatitis C virus seroconversion. The time from initial hepatitis C virus detection by hepatitis C virus core antigen assay and anti-hepatitis C virus seroconversion was not >4 weeks. De novo acquisition of hepatitis C virus in haemodialysis was associated with a rise in alanine aminotransferase levels. Hepatitis C virus core antigen enzyme-linked immunosorbent assay test results can be obtained in routine laboratories without the need of special equipment or training. Hepatitis C virus core antigen testing among anti-hepatitis C virus negative patients on maintenance dialysis is suggested in order to early assess de novo hepatitis C virus within dialysis units. PMID- 15801922 TI - A phase 2 clinical trial of metformin as a treatment for non-diabetic paediatric non-alcoholic steatohepatitis. AB - BACKGROUND: Children with non-alcoholic steatohepatitis are insulin-resistant and metformin has been proposed as a potential therapy. However, paediatric safety and efficacy data are absent. AIM: To test the hypothesis that metformin therapy will safely improve markers of liver disease in paediatric non-alcoholic steatohepatitis. METHODS: Single-arm open-label pilot study of metformin 500 mg twice daily for 24 weeks in non-diabetic children with biopsy-proven non alcoholic steatohepatitis. RESULTS: Ten obese children (mean body mass index 30.4) enrolled and completed the trial. Mean alanine aminotransferase and aspartate aminotransferase (AST) improved significantly (P < 0.01) from baseline (184, 114 U/L) to end of treatment (98, 68 U/L). Alanine aminotransferase normalized in 40% and AST normalized in 50% of subjects. Children demonstrated significant improvements in liver fat measured by magnetic resonance spectroscopy (30-23%, P < 0.01); insulin sensitivity measured by quantitative insulin sensitivity check index (0.294-0.310, P < 0.05); and quality of life measured by pediatric quality of life inventory 4.0 (69-81, P < 0.01). CONCLUSION: Open-label treatment with metformin for 24 weeks was notable for improvement in liver chemistry, liver fat, insulin sensitivity and quality of life. A large randomized controlled trial is needed to definitively determine the efficacy of metformin for paediatric non-alcoholic steatohepatitis. PMID- 15801923 TI - Anti-Saccharomyces cerevisiae and perinuclear anti-neutrophil cytoplasmic antibodies in coeliac disease before and after gluten-free diet. AB - BACKGROUND: Anti-Saccharomyces cerevisiae and perinuclear anti-neutrophil cytoplasmic autoantibodies are markers of Crohn's disease and ulcerative colitis respectively. AIM: To determine the prevalence of anti-S. cerevisiae and perinuclear anti-neutrophil cytoplasmic autoantibodies in a large series of coeliac disease patients before and after gluten free diet, and to correlate anti S. cerevisiae-positivity with intestinal mucosal damage. METHODS: One hundred and five consecutive coeliac disease patients and 141 controls (22 ulcerative colitis, 24 Crohn's disease, 30 primary sclerosing cholangitis, 15 postenteritis syndrome, 50 blood donors) were tested for anti-S. cerevisiae by enzyme-linked immunosorbent assay and for perinuclear anti-neutrophil cytoplasmic autoantibodies by indirect immunofluorescence. RESULTS: In coeliac disease anti S. cerevisiae (immunoglobulin G and/or immunoglobulin A) were slightly less frequent (59%) than in Crohn's disease (75%, P = 0.16) and significantly more frequent than in ulcerative colitis (27%), primary sclerosing cholangitis (30%), postenteritis syndrome (26%) and blood donors (4%) (P = 0.009, P = 0.0002, P = 0.025, P < 0.0001). No correlation was found between anti-S. cerevisiae and degree of mucosal damage. Perinuclear anti-neutrophil cytoplasmic autoantibodies were detected only in one coeliac. After gluten free diet the disappearance of anti-S. cerevisiae-immunoglobulin A (93%) was more frequent than that of immunoglobulin G (17%, P = 0.0001); perinuclear anti-neutrophil cytoplasmic autoantibodies disappeared in the only coeliac positive at diagnosis. CONCLUSION: More than half of untreated coeliacs are anti-S. cerevisiae-positive irrespective of the severity of mucosal damage. Differently from immunoglobulin A, anti-S. cerevisiae-immunoglobulin G persisted in more than 80% after gluten free diet. The high prevalence of anti-S. cerevisiae in coeliac disease suggests that they may be the effect of a non-specific immune response in course of chronic small bowel disease. PMID- 15801924 TI - The out-patient management of patients with acute mild-to-moderate colonic diverticulitis. AB - BACKGROUND: There are no management criteria for optimum out-patient care in mild to-moderate acute colonic diverticulitis. AIM: To enable such patients to be managed in an out-patient setting, by establishing criteria and treatment protocols. METHODS: We conducted an open trial and follow-up study from 1997 to 2002. On the basis of ultrasonography, we defined and categorized mild-to moderate acute colonic diverticulitis ranging from limited inflammation within diverticulum to an abscess < 2 cm in diameter. Subjects were treated as out patients and followed a 10-day treatment protocol consisting of an oral antibiotic and a sports drink for the first 3 days. Physical examination and laboratory testing helped determine whether or not a patient could resume a liquid diet on day 4, and a regular diet on day 7. RESULTS: Of the 70 patients, 68 were successfully treated. Two patients required hospitalization. Of the 65 patients who were tracked over several months [median (intraquarter range) = 30.8 (11.9-44.2) months], 16 had one or more clinical recurrences. The medical cost per episode was 80% lower than in-patient treatment. CONCLUSIONS: Patients with mild-to-moderate acute colonic diverticulitis can be safely and successfully treated as out-patients using this protocol. PMID- 15801925 TI - A randomized, double blind, comparative trial of micafungin (FK463) vs. fluconazole for the treatment of oesophageal candidiasis. AB - AIM: To determine efficacy and safety of intravenous micafungin vs. intravenous fluconazole in the treatment of oesophageal candidiasis. METHODS: A total of 523 patients > or =16 years with documented oesophageal candidiasis were randomized (1:1) in this controlled, non-inferiority study to receive either micafungin (150 mg/day) or fluconazole (200 mg/day). Response was evaluated clinically and endoscopically. Post-treatment assessments were performed at 2 and 4 weeks after discontinuation of therapy. RESULTS: Median duration of therapy was 14 days. For the primary end-point of endoscopic cure, treatment difference was -0.3% (micafungin 87.7%, fluconazole 88.0%). Documented persistent invasive disease at the end of therapy was reported in 2.7% and 3.9% of patients, respectively. Both 84.8% of micafungin and 88.7% of fluconazole patients remained recurrence free at 4-weeks post-treatment. The overall therapeutic response rate was 87.3% for micafungin and 87.2% for fluconazole. The incidence of drug-related adverse events was 27.7% for micafungin and 21.3% for fluconazole. Six (2.3%) micafungin- and two (0.8%) fluconazole-treated patients discontinued therapy; rash was the most common event leading to discontinuation. CONCLUSION: Intravenous micafungin (150 mg daily) is well tolerated and as efficacious as intravenous fluconazole (200 mg daily) in the primary treatment of oesophageal candidiasis, achieving high rates of clinical and endoscopic cure. PMID- 15801926 TI - Helicobacter pylori duodenal colonization is a strong risk factor for the development of duodenal ulcer. AB - AIM: To test the hypothesis that duodenal colonization represents the final crucial step in the development of Helicobacter pylori related duodenal ulcer. METHODS: Patients with non-ulcer dyspepsia who had gastric colonization by H. pylori were included in the study. At baseline endoscopy, we evaluated the prevalence of duodenal colonization (culture, urease testing and histology), and cytotoxin-associated gene A status (polymerase chain reaction). No patients received eradication during 1 year follow-up. At this time, endoscopy was repeated and the incidence of duodenal ulcer was assessed. RESULTS: Among 181 patients completing follow-up, 53 (29%) had duodenal colonization: 72% of them were cytotoxin-associated gene A positive, versus 37% patients without duodenal colonization (P < 0.001). Duodenal ulcer developed in 12 (22.6%) patients with duodenal colonization and in two (1.6%) without duodenal colonization (odds ratio for duodenal ulcer: 6.29, 95% confidence intervals 2.44-17.45). The incidence of duodenal ulcer was similar among cytotoxin-associated gene A positive and cytotoxin-associated gene A negative subjects with duodenal colonization: 21.05% versus 26.6%. CONCLUSIONS: The assessment of duodenal colonization by H. pylori in patients with non-ulcer dyspepsia is strongly predictive for the subsequent development of duodenal ulcer and may help to stratify patients at risk for this disease. PMID- 15801927 TI - Oesophageal pH has a power-law distribution in control and gastro-oesophageal reflux disease subjects. PMID- 15801928 TI - Infliximab and the bone in Crohn's disease. PMID- 15801930 TI - Drug development: more science, more education. PMID- 15801931 TI - Measuring bronchodilation in COPD clinical trials. AB - AIMS: The aim of this study was to compare the variability and sensitivity of impulse oscillometry (R5, X5 and RF), plethysmography (Raw and sGaw) and spirometry (FEV1, FVC and MMEF) in order to determine the most powerful technique for assessing bronchodilation in COPD clinical trials. METHODS: Twenty-four patients with COPD had impulse oscillometry, plethysmography and spirometry measured twice 30 mins apart, to determine variability. Then ascending doses of salbutamol (20, 50, 100, 200, 400 and 800 microg) were given and the same measurements made after each dose. Significant changes greater than variability were determined for each performed measurement (expressed as mean percentage improvement with 95% CI). RESULTS: Significant effects (P < 0.05) were detected after 20 microg by X5 (18.5% CI 9.8-27.2) RF (11.1% CI 7.2-15.0) and sGaw (21.5% CI 10.1-32.9), and after 50 microg by R5 (16.7% CI 10.8-22.5) and Raw (19.7% CI 13.0-26.4). FEV1 was less sensitive, detecting significant bronchodilation at 100 microg (10.2% CI 7.4-12.9). CONCLUSIONS: We conclude that impulse oscillometry and plethysmography should be considered the preferred techniques for measuring bronchodilation in COPD clinical trials. PMID- 15801932 TI - Quantifying bronchodilator responses in chronic obstructive pulmonary disease trials. PMID- 15801933 TI - Reproducibility of forearm vasodilator response to intra-arterial infusion of calcitonin gene-related peptide assessed by venous occlusion plethysmography. AB - AIMS: To assess the reproducibility of the forearm blood flow (FBF) response to intra-arterial infusion of calcitonin-gene related peptide (CGRP), measured by venous occlusion plethysmography. In addition, to compare different ways of expressing the FBF response and perform sample size calculations. METHODS: On two separate visits, CGRP (10 ng min(-1) dl(-1) forearm) was infused for 45 min into the brachial artery of six healthy subjects. Reproducibility was assessed by calculating mean difference, repeatability coefficient, within-subject coefficient of variation (WCV) and intraclass correlation coefficient. RESULTS: CGRP increased FBF from 2.8 +/- 0.4 and 3.2 +/- 0.7 (at baseline) to 15.4 +/- 1.4 and 15.2 +/- 1.5 ml min(-1) dl(-1) forearm (at 45 min) on visits 1 and 2, respectively (P < 0.0001 for both visits). Mean difference in FBF at 45 min between both visits was 0.3 ml min(-1) dl(-1) forearm (repeatability coefficient: 4.1 ml min(-1) dl(-1) forearm). This FBF response appeared to be more reproducible when expressed as absolute FBF in the infused arm (WCV 11%) compared with absolute FBF-ratio between both arms (WCV 37%), percentage change from baseline in FBF in the infused arm (WCV 29%) and percentage change from baseline in FBF-ratio (WCV 40%). When expressed as absolute FBF, a sample size of five (95% confidence interval: 2-12) subjects gives 90% power at a type I error probability of 0.05 to detect a 25% shift in FBF response. CONCLUSIONS: Intra arterial infusion of CGRP results in a forearm vasodilator response which is reproducible between days. This response is most reproducible when expressed as absolute FBF. The presented methodology provides a suitable pharmacodynamic model to assess the in vivo activity of CGRP-receptor antagonists in a small number of subjects. PMID- 15801934 TI - Absence of circadian variation in the pharmacokinetics of lopinavir/ritonavir given as a once daily dosing regimen in HIV-1-infected patients. AB - AIMS: To compare the pharmacokinetics of lopinavir/ritonavir (LPV/r) 800/200 mg administered once daily in the morning compared with the evening. METHODS: This was a randomized, two-way, cross-over study in HIV+ subjects. In each subject the pharmacokinetics of each drug were characterized after 2 weeks of LPV/r 800/200 mg administered once daily at 08.00 h and 19.00 h. On study days, LPV/r was taken with a standardized meal (800 kCal, 25% from fat) after fasting for at least 5 h. LPV/r concentrations were measured by LC-MS/MS, and the data were analyzed by noncompartmental pharmacokinetic analysis. RESULTS: Fourteen subjects completed the study (all men, mean age/weight 44 year/81 kg). The median (interquartile range) LPV AUC(0,24 h), maximum plasma concentration (C(max)) and concentration at the end of the dosing interval (C(24 h)) after am and pm dosing was, respectively, 143 (116-214) mg l(-1) h, 12.8 (10.3-17.2) mg l(-1), 1.34 (0.58 3.25) mg l(-1), and 171 (120-232) mg l(-1) h, 12.9 (8.22-16.3) mg l(-1), 1.15 (0.59-1.98) mg l(-1). The geometric mean ratio (GMR, am : pm) and 95% CI of the LPV AUC(0,24 h), C(max), and C(24 h) was 0.91 (0.79, 1.06), 1.11 (0.94, 1.32), and 1.19 (0.72, 1.96), respectively. The median ritonavir C(max) after am and pm dosing was 1.05 and 0.90 mg l(-1), respectively. The GMR (95% CI) of the RTV AUC(0,24 h), C(max), and C(24 h) was 0.93 (0.80, 1.08), 1.27 (1.00, 1.63), and 1.04 (0.68, 1.60), respectively. Administration of LPV/r in a once-daily regimen was generally well tolerated. CONCLUSIONS: No differences were observed in the pharmacokinetics of LPV/r after am or pm dosing with food, which suggests that this once daily combination, can be taken in the morning or evening. Such flexibility in dosing may improve adherence. PMID- 15801935 TI - Steady-state pharmacokinetics and pharmacodynamics of CHF3381, a novel antineuropathic pain agent, in healthy subjects. AB - AIMS: To evaluate the safety, tolerability, pharmacokinetic and pharmacodynamic profiles of CHF3381, a dual NMDA and MAO-A inhibitor, after multiple oral doses in healthy subjects. METHODS: Forty-eight young males received CHF3381 at doses of 100 mg twice daily, 200 mg twice daily, 400 mg twice daily or placebo for 2 weeks according to a double-blind, randomized, parallel group design. Plasma and urine concentrations of the parent drug and of two major metabolites (CHF3567 and 2-aminoindane) were measured over time. MAO-A activity in plasma was estimated by measuring plasma concentrations of 3,4-dihydroxyphenylglycol. Sustained attention, memory and sedation were assessed throughout the study with standard psychometric tests. RESULTS: Most of the adverse events were mild in intensity, with dose regimens of 100 mg twice daily and 200 mg twice daily being indistinguishable from placebo. After 400 mg twice daily, the most frequent adverse events were mild dizziness, asthenia and insomnia. At steady-state, 400 mg twice daily slightly increased supine heart rate (+ 9 +/- 2 beats min(-1)) and diastolic blood pressure (+6 +/- 2 mmHg) compared with placebo. There were no dose-dependent or consistent effects of CHF3381 on attention, motor co-ordination or memory, but 400 mg twice daily significantly decreased alertness compared with placebo. Plasma concentrations of CHF3381 peaked at around 3 h and were dose proportional. The elimination half-life of CHF3381 was estimated to be 4-6 h. At steady-state, significant CHF3381 plasma concentrations were detected at predose with a modest accumulation (1.3-1.5 times), showing that the drug given twice daily is active over the entire 24 h period. Plasma concentrations of CHF3567 and of 2-aminoindane were also proportional to the dose of CHF3381. CHF3381 dose dependently inhibited MAO-A activity with peak effects at steady-state of 27 +/- 4%, 46 +/- 2% and 65 +/- 5% after 100 mg twice daily, 200 mg twice daily and 400 mg twice daily, respectively. There were no significant effects of CHF3381 on attention (rapid visual information processing), motor co-ordination (body sway) or memory (learning memory task) at any of the doses. At steady-state, there was a significant decrease in alertness (Bond & Lader visual analogue scale) in the 400 mg twice daily group compared with placebo. CONCLUSIONS: A twice daily regimen of CHF3381 appears to be adequate from a pharmacokinetic and pharmacodynamic perspective. Plasma concentrations reached with 400 mg twice daily exceeded those observed in animals receiving pharmacologically active doses in chronic pain models. PMID- 15801936 TI - Pharmacogenetic profiling across the irinotecan pathway in Asian patients with cancer. AB - AIMS: The aim of this exploratory study was to investigate associations between irinotecan pharmacokinetic parameters and allelic variants in genes encoding for drug transporters and drug metabolizing enzymes that are involved in irinotecan disposition in Asian patients with cancer. METHODS: Irinotecan was administered at 100 mg m(-2) over 90 min on a weekly schedule to 29 nasopharyngeal carcinoma patients and pharmacokinetic analysis was performed during the first cycle. All patients were genotyped for allelic variants in genes encoding drug metabolizing enzymes (CYP3A4, CYP3A5, UGT1A1) and drug transporters (ABCB1, ABCC2 and ABCG2) that are involved in irinotecan disposition. RESULTS: Of the six candidate genes that were analyzed, 11 genetic variants were found. Significant genotypic phenotypic associations were apparent only for transporter genes. The C(max) of irinotecan was significantly lower in patients carrying the CC genotype at exon 26 of the ABCB1 gene compared with those harbouring at least one variant allele (P = 0.047). Patients harbouring the wild type ABCG2 CTCA genotype were associated with significantly higher values for relative extent of conversion (REC) of irinotecan to SN-38 compared with patients carrying at least one deletion CTCA allele (P = 0.019). CONCLUSIONS: The present exploratory study shows that genetic polymorphisms in drug transporter genes, particularly in ABCB1 and ABCG2 genes, may be important in influencing the pharmacokinetics of irinotecan and its metabolites. The predictive value of the identified allelic variants in the ABCG2 and ABCB1 genes on irinotecan disposition should be further investigated in a larger patient population as well as in other ethnic populations. PMID- 15801937 TI - Effect of ginkgo and ginger on the pharmacokinetics and pharmacodynamics of warfarin in healthy subjects. AB - AIM: The aim of this study was to investigate the effect of two common herbal medicines, ginkgo and ginger, on the pharmacokinetics and pharmacodynamics of warfarin and the independent effect of these herbs on clotting status. METHODS: This was an open label, three-way crossover randomized study in 12 healthy male subjects, who received a single 25 mg dose of warfarin alone or after 7 days pretreatment with recommended doses of ginkgo or ginger from herbal medicine products of known quality. Dosing with ginkgo or ginger was continued for 7 days after administration of the warfarin dose. Platelet aggregation, international normalized ratio (INR) of prothrombin time, warfarin enantiomer protein binding, warfarin enantiomer concentrations in plasma and S-7-hydroxywarfarin concentration in urine were measured. Statistical comparisons were made using anova and the 90% confidence intervals (CIs) of the ratio of log transformed parameters are reported. RESULTS: INR and platelet aggregation were not affected by administration of ginkgo or ginger alone. The mean (95% CI) apparent clearances of S-warfarin after warfarin alone, with ginkgo or ginger were 189 (167-210) ml h(-1), 200 (173-227) ml h(-1) and 201 (171-231) ml h(-1), respectively. The respective apparent clearances of R-warfarin were 127 (106-149) ml h(-1), 126 (111-141) ml h(-1) and 131 (106-156) ml h(-1). The mean ratio (90% CI) of apparent clearance for S-warfarin was 1.05 (0.98-1.21) and for R-warfarin was 1.00 (0.93-1.08) when coadministered with ginkgo. The mean ratio (90% CI) of AUC(0-168) of INR was 0.93 (0.81-1.05) when coadministered with ginkgo. The mean ratio (90% CI) of apparent clearance for S-warfarin was 1.05 (0.97-1.13) and for R-warfarin was 1.02 (0.95-1.10) when coadministered with ginger. The mean ratio (90% CI) of AUC(0-168) of INR was 1.01 (0.93-1.15) when coadministered with ginger. The mean ratio (90% CI) for S-7-hydroxywarfarin urinary excretion rate was 1.07 (0.85-1.32) for ginkgo treatment, and 1.00 (0.81-1.23) for ginger coadministration suggesting these herbs did not affect CYP2C9 activity. Ginkgo and ginger did not affect the apparent volumes of distribution or protein binding of either S-warfarin or R-warfarin. CONCLUSIONS: Ginkgo and ginger at recommended doses do not significantly affect clotting status, the pharmacokinetics or pharmacodynamics of warfarin in healthy subjects. PMID- 15801938 TI - Effect of gemfibrozil on the pharmacokinetics and pharmacodynamics of racemic warfarin in healthy subjects. AB - AIMS: Case reports suggest that gemfobrozil can increase the anticoagulant effect of warfarin. Because gemfibrozil inhibits CYP2C9 in vitro, we studied its effects on the pharmacokinetics and pharmacodynamics of racemic warfarin. METHODS: In a randomized cross-over study, 10 healthy subjects ingested 600 mg gemfibrozil or placebo twice daily for 8 days. On day 3, they were administered a single dose of 10 mg racemic R-S-warfarin orally. The concentrations of R- and S-warfarin in plasma and thromboplastin time were monitored up to 168 h. RESULTS: Gemfibrozil decreased the mean (+/-SD) area under the plasma concentration-time curve [AUC((0 infinity))] of S-warfarin by 11%, from 19.9 +/- 5.2 mg l(-1) h to 17.6 +/- 4.7 mg l(-1) h (95% CI on the difference -3.7, -0.78; P < 0.01) and that of R-warfarin by 6% from 31.3 +/- 7.5 mg l(-1) h during the gemfibrozil phase to 29.5 +/- 6.9 mg l(-1) h during the placebo phase (95% CI -3.3, -0.33; P < 0.05). There were no significant differences in the elimination half-lives of S- or R-warfarin between the phases. Gemfibrozil did not alter the anticoagulant effect of warfarin. CONCLUSION: Unexpectedly, gemfibrozil slightly decreased the plasma concentrations of R- and S-warfarin. Displacement of warfarin from plasma albumin by gemfibrozil or its interference with the absorption of warfarin could explain the present findings. Usual therapeutic doses of gemfibrozil seem to have limited effects on the pharmacokinetics and pharmacodynamics of single dose warfarin in healthy subjects. PMID- 15801940 TI - Self treatment with one of three self selected, ultramolecular homeopathic medicines for the prevention of upper respiratory tract infections in children. A double-blind randomized placebo controlled trial. AB - AIMS: Homeopathic medicines are frequently purchased over the counter (OTC). Respiratory complaints are the most frequent reason for such purchases. Children with upper respiratory tract infection (URTI) are frequent users of homeopathy. This study investigates the effect of self treatment with one of three self selected ultramolecular homeopathic medicines for the prevention of childhood URTI. METHODS: A double-blind randomized parallel group placebo controlled trial was carried out in 251 children below the age of 10 years, recruited by post from those previously diagnosed with URTI when attending a casualty department. The children were randomly assigned to receive either placebo or ultramolecular homeopathic medicines in C-30 potency (diluted 10(-60)) administered twice weekly for 12 weeks. Parents chose the medicine based on simplified constitutional indications for the three medicines most frequently prescribed by Norwegian homeopaths for this group of patients. The main outcome measure relates to the prevention of new episodes of URTI measured with median total symptom score over 12 weeks. RESULTS: There was no difference in the predefined primary outcome between the two groups (P = 0.733). Median URTI scores over 12 weeks in the homeopathic medicine group were 26.0 (95% confidence interval (CI) 16.3, 43.7) and for placebo 25.0 (95% CI 14.2, 38.4). There was no statistical difference between the two groups in median number of days with URTI symptoms or in the use of conventional medication/care. CONCLUSIONS: In this study there was no effect over placebo for self treatment with one of three self selected, ultramolecular homeopathic medicines in preventing childhood URTI. This can be due to the lack of effect of the highly diluted homeopathic medicines or the process of selection and type of medicines. PMID- 15801939 TI - Diltiazem inhibits human intestinal cytochrome P450 3A (CYP3A) activity in vivo without altering the expression of intestinal mRNA or protein. AB - AIMS: To determine the effect of diltiazem on intestinal CYP3A activity and protein and mRNA expression in vivo in healthy subjects. METHODS: Intestinal biopsies were obtained from ten healthy controls and from ten healthy subjects after receiving diltiazem 120 mg bid for 7 days. Intestinal CYP3A activity, CYP3A4 protein and mRNA concentrations were quantified in both groups. Intestinal CYP3A activity was determined by incubation of small bowel homogenate with midazolam (25 microM) and NADPH for 5 min and the rate of formation of 1' hydroxymidazolam was quantified. RESULTS: All subjects in the treatment group had detectable diltiazem concentration in the serum. While there was no significant difference in CYP3A4 protein and mRNA expression between the control and treatment groups, the formation of 1'-hydroxymidazolam (446 pmol min(-1) mg(-1) 6 (control) vs. 170 (CI 112, 228) pmol min(-1) mg(-1) 95% confidence interval (CI 269, 623) (diltiazem group)) was significantly reduced (P < 0.05). CONCLUSION: Diltiazem decreased small bowel CYP3A activity by 62% as a result of irreversible inhibition with no corresponding change in intestinal CYP3A4 mRNA or protein concentrations. PMID- 15801941 TI - Ineffectiveness of lipid-lowering therapy in primary care. AB - BACKGROUND: Evidence confirms the positive effects of lipid-lowering agents on the risk of cardiovascular disease. Local guidelines in France (AFSSAPS) have defined therapeutic objectives for LDL-cholesterol. These objectives vary with the number of cardiovascular risk factors in addition to dyslipidaemia. We determined the proportions of patients at therapeutic objective in different classes of cardiovascular risk to test the hypothesis that compliance with guidelines varies across the levels of risk. Comparison with international guidelines (ANDEM) was also performed. METHODS: A group of 3173 dyslipidaemic patients treated with lipid-lowering agents and managed by general practitioners was randomly selected from BKL-Thales panel, a French computerized database. For each patient, history of coronary heart disease and the number of cardiovascular risk factors were documented. Compliance with guidelines was assessed from achievement of therapeutic objective. RESULTS: The study population included 79% primary prevention patients (1.6, 25.5, 31.7 and 20.1%, with 1, 2, 3, and >3 risk factors, respectively) and 21.0% secondary prevention patients. Applying AFSSAPS guidelines, the proportions of primary prevention patients not at LDL-cholesterol objectives varied across risk categories (P < 0.0001), from 3.9% for patients with one risk factor to 46.5% for patients with >3 risk factors, and therapeutic failure reached 39.9% in secondary prevention. Only 26% of patients who were at high cardiovascular risk (>3 risk factors or prior coronary heart disease) and not at therapeutic objective received high doses (>standard recommended doses) of lipid-lowering agents in monotherapy. Applying ANDEM guidelines, 74% of secondary prevention patients were not at treatment goal. CONCLUSION: Compliance with guidelines varied inversely with the level of cardiovascular risk. Besides, most patients not at therapeutic objective were not up-titrated. The use of lipid lowering agents is inadequate, depriving many patients of an effective protection against cardiovascular diseases. PMID- 15801942 TI - The teaching of drug development to medical students: collaboration between the pharmaceutical industry and medical school. AB - Collaboration between the medical school at Leicester and a local pharmaceutical company, AstraZeneca, led to the design and implementation of an optional third year special science skills module teaching medical students about drug discovery and development. The module includes didactic teaching about the complexities of the drug discovery process leading to development of candidate drugs for clinical investigation as well as practical experience of the processes involved in drug evaluation preclinically and clinically. It highlights the major ethical and regulatory issues concerned with the production and testing of novel therapies in industry and the NHS. In addition it helps to reinforce other areas of the medical school curriculum, particularly the understanding of clinical study design and critical appraisal. The module is assessed on the basis of a written dissertation and the critical appraisal of a drug advertisement. This paper describes the objectives of the module and its content. In addition we outline the results of an initial student evaluation of the module and an assessment of its impact on student knowledge and the opinion of the pharmaceutical industry partner. This module has proven to be popular with medical students, who acquire a greater understanding of the work required for drug development and therefore reflect more favourably on the role of pharmaceutical companies in the UK. PMID- 15801943 TI - Therapeutic innovation in the European Union: analysis of the drugs approved by the EMEA between 1995 and 2003. AB - Since January 1995, all European Union applications for marketing approval for medicinal products derived from biotechnology and other drugs considered potentially innovative follow the EMEA centralized procedure. In order to assess the overall degree of therapeutic innovation of these drugs, we considered, for each approved agent, its target, the availability of previous treatments and the extent of its therapeutic effect. The following scores for therapeutic innovation were assigned through a consensus process: 'A' (important), 'B' (moderate) and 'C' (modest). The overall degree of important/moderate therapeutic innovation was 47% of all therapeutic agents (32% important; 15% moderate). Most (80%) of the EMEA-approved therapeutic agents were for serious diseases. The remaining ones were for risk factors (7%) or nonserious diseases (13%). PMID- 15801944 TI - Raloxifene pharmacokinetics in males with normal and impaired renal function. AB - AIMS: To compare raloxifene pharmacokinetics between renally impaired and healthy subjects. METHODS: Raloxifene 120 mg was administered to 10 males with renal impairment (creatinine 2-4 mg dl(-1)) and to 10 healthy males. Data were analysed by two noncompartmental and one compartmental nonlinear regression methods. RESULTS: The medians (95% confidence interval) of the area under the curves (AUC) were 35.1 (25.8, 74) and 20.5 (16.8, 28.0) h ng ml(-1) per mg kg(-1), P < 0.01, and of the clearances (CL/F) were 28.5 (13.5, 38.8) and 48.8 (35.8, 59.4) l h(-1) kg(-1), P < 0.01, in renally impaired and healthy subjects, respectively. 95% Confidence intervals on the differences for AUC and CL/F were 6.5-44.1 and -35.1 to -7.9, respectively. CONCLUSION: Exposure to raloxifene was twice as high in males with renal impairment compared with healthy subjects. PMID- 15801945 TI - Insertion/deletion polymorphism of the ACE gene and adherence to ACE inhibitors. AB - AIMS: We investigated whether the insertion/deletion (I/D) polymorphism of the ACE gene modified the adherence to ACE inhibitors as measured by the discontinuation of an ACE inhibitor, or addition of another antihypertensive drug. METHODS: This was a cohort study among 239 subjects who started ACE inhibitor therapy. A Cox proportional hazard model was used to calculate relative risk (RR). RESULTS: During follow-up there was no significant difference between subjects with the DD, ID or II genotype (DD vs II; RR = 1.17, 95%CI: 0.78, 1.77 and ID vs II; RR = 1.06, 95%CI: 0.73, 1.55) in adherence. CONCLUSIONS: The I/D polymorphism of the ACE gene does not influence the adherence to ACE inhibitors. PMID- 15801946 TI - Co-administration of oxycodone and morphine and analgesic synergy re-examined. PMID- 15801948 TI - Different accumulation of temocapril and enalapril during repeated dosing in elderly patients. PMID- 15801949 TI - Scleromalacia perforans complicating graft-versus-host disease. PMID- 15801950 TI - Sinus zygomycosis in a patient receiving voriconazole prophylaxis. PMID- 15801951 TI - The aetiology and management of Castleman disease at 50 years: translating pathophysiology to patient care. AB - Fifty years ago, Dr Benjamin Castleman first described the unusual lymphoproliferative disorder that now bears his name. Over the subsequent decades, astute clinical and pathologic observations coupled with clever molecular biologic research have increased our understanding of the aetiology of Castleman disease (CD). This article proposes three broad CD variants based on both distinctive histopathology and clinical behaviour. The pivotal roles of infection with human herpesvirus 8 and interleukin-6 production in the development of CD are emphasized. Finally, the natural history of CD and the myriad of therapeutic options are reviewed in the context of a unified model of CD pathophysiology, and continued areas of uncertainty are discussed. PMID- 15801952 TI - Biology and management of relapsed acute myeloid leukaemia. AB - Disease relapse remains the major cause of treatment failure in adults with acute myeloid leukaemia (AML). This reflects both the failure of current salvage regimens and the absence of effective strategies to secure long-term disease-free survival in those patients who achieve a second remission. Recent progress in understanding the pathogenesis of relapsed disease has enabled the identification of a variety of dysregulated molecular pathways and these now provide a rational basis for the design of novel targeted therapies. At the same time, advances in allogeneic stem-cell transplantation have permitted the extension of the curative potential of allografting to patients in whom it was previously contraindicated. As a result, a range of novel drug and transplant therapies has become available in patients with relapsed AML, and it is realistic to anticipate that a co ordinated assessment of their clinical and biological impact will provide the basis for the design of future, more effective treatments in relapsed disease. PMID- 15801953 TI - Early response to induction is predictive of survival in childhood Philadelphia chromosome positive acute lymphoblastic leukaemia: results of the Medical Research Council ALL 97 trial. AB - We report on the outcome of children with Philadelphia positive acute lymphoblastic leukaemia (Ph+ ALL) treated on the UK Medical Research Council (MRC) trial for childhood ALL, MRC ALL 97, between January 1997 and June 2002. Forty-two (2.3%) patients were Ph+. Nineteen (45%) had <25% blasts in bone marrow (BM) within the first 2 weeks of treatment and were defined as a good response group (GRG), the others as a poor response group (PRG). Thirty-six (86%) achieved first complete remission (CR1) at the end of induction, of which 28 underwent BM transplantation (BMT). The median follow-up was 42 months (range, 21-84). The 3 year event-free survival (EFS; 52%, 95% CI, 36-66%) was a considerable improvement on the previous MRC UKALL XI trial (27%). EFS for the GRG and PRG were 68% (43-84%) and 39% (18-59%), respectively (P = 0.03); presenting white cell count <50 x 10(9)/l (P = 0.02) was predictive for overall survival. Changes in the MRC ALL97 trial within the study period resulted in some Ph+ ALL receiving daunorubicin and either prednisolone or dexamethasone during induction. Though the use of daunorubicin during induction was not a prospective study question, EFS was significantly better for those whose induction included this drug (P = 0.02). Steroid randomization was not stratified for Ph+ ALL patients and was not predictive for EFS. BMT in CR1 appeared to reduce the risk of a subsequent BM relapse. These results show significant improvement on previous MRC trials; future therapeutic strategies should include early intensive therapy and BMT in CR1. PMID- 15801954 TI - Interphase FISH to detect PBX1/E2A fusion resulting from the der(19)t(1;19)(q23;p13.3) or t(1;19)(q23;p13.3) in paediatric patients with acute lymphoblastic leukaemia. AB - Approximately 6% of paediatric patients with precursor B-cell acute lymphoblastic leukaemia (B-ALL) harbour a rearrangement involving the gene regions of PBX1 (1q23) and E2A (19p13.3) which is visualized cytogenetically either as a der(19)t(1;19)(q23;p13.3) or the less common balanced t(1;19)(q23;p13.3). Unfortunately, no commercial dual-colour, double fusion fluorescence in situ hybridization (D-FISH) strategies are available to detect this recurrent anomaly. Therefore, we have created a D-FISH assay to detect these translocations and monitor minimal residual disease. This probe set was created using four bacterial artificial chromosomes (BACs) corresponding to the PBX1 gene region at 1q23 and four BACs corresponding to the E2A gene region at 19p13.3. We analysed 30 negative bone marrow controls and 20 diagnostic and post-treatment specimens from 13 paediatric B-ALL patients with a cytogenetically defined 1;19 translocation. Once unblinded, the results demonstrated that our D-FISH method effectively identified all diagnostic samples as abnormal and identified disease in four post treatment samples that were previously considered to be normal by conventional cytogenetic analysis. The development of this FISH strategy for the detection of der(19)t(1;19)(q23;p13.3) and t(1;19)(q23;p13.3) proved to be an effective technique, allowing both the detection of disease in diagnostic samples and in post-treatment samples. PMID- 15801955 TI - p53 Aberrations do not predict individual response to fludarabine in patients with B-cell chronic lymphocytic leukaemia in advanced stages Rai III/IV. AB - Abnormalities of p53 have been associated with short survival and non-response to therapy in chronic lymphocytic leukaemia (CLL). We have evaluated the rate of response to fludarabine as first-line therapy in 54 patients with advanced stage CLL, analysing the cytogenetic profile, aberrations in p53, including the methylation status of its promoter, and the immunoglobulin heavy-chain variable region (IGVH) mutation status. According to the advanced stage of the disease in this series, 75% of patients presented genetic aberrations associated with poor prognosis: del(17p) and/or del(11q), and no-mutated IGVH genes. Ten patients (18.5%) had methylation in the promoter region of p53. Eighty-three per cent of patients treated achieved a response, with a high rate of complete remission (47.6%). Although we found a significant correlation between failures and the presence of p53 aberrations (P = 0.0065), either with methylation (P = 0.018) or deletion (P = 0.015), 64% of the patients with aberrations in this gene responded to treatment (11/17), suggesting that fludarabine induces high remission rates, even in these patients. This is the first time that the significance of p53 promoter methylation status is described in this pathology, and our data support that this epigenetic phenomenon could be involved in the pathogenesis and clinical evolution of CLL. PMID- 15801956 TI - Aberrant methylation of the negative regulators RASSFIA, SHP-1 and SOCS-1 in myelodysplastic syndromes and acute myeloid leukaemia. AB - Mutations in the receptor tyrosine kinase (RTK/RAS) signalling pathway frequently provide a proliferative signal in myeloid malignancies. However, the role of RASSF1A, SHP-1 and SOCS-1, negative regulators of RTK/RAS signalling, has not been extensively investigated in the myelodysplastic syndromes (MDS) or acute myeloid leukaemia (AML). This study employed methylation-specific polymerase chain reaction (MS-PCR) to determine if aberrant promotor methylation of RASSF1A, SHP-1 and SOCS-1 is involved in the pathogenesis of myeloid malignancies. Patients with MDS (n = 107), AML (n = 154) and juvenile myelomonocytic leukaemia (JMML, n = 5) were investigated, together with 15 normal controls. Primers were located in the promotor region of each gene as well as within exon 2 of SOCS-1. Methylation of RASSF1A was found in five of 55 (9%) MDS cases, but not in any of 57 AML cases studied. RASSF1A methylation was present in one case (20%) of JMML. SHP-1 methylation was present in 13 of 121 (11%) AML cases but was not found in MDS or JMML. SOCS-1 promoter methylation was present in eight of 74 (11%) MDS patients but was not seen in JMML or AML. Importantly, RAS mutations and RASSF1A and SOCS-1 methylation were mutually exclusive indicating that approximately 30% of MDS cases had a defect of the RTK/RAS pathway and its negative regulation. Finally, SOCS-1 exon 2 methylation may not be pathogenetically relevant, since it was detected in samples from normal individuals and did not correlate with promotor methylation. PMID- 15801957 TI - Gains on 9p are common genomic aberrations in idiopathic myelofibrosis: a comparative genomic hybridization study. AB - Ideopathic myelofibrosis (IMF) is a chronic myeloproliferative disorder resulting in bone marrow fibrosis as a consequence of growth factor release from clonal haematopoiesis. Conventional cytogenetic analysis identifies abnormalities in approximately a third of cases at diagnosis, although rarely uncovers unique, primary genetic events. We have used comparative genomic hybridization (CGH) to study 25 IMF cases and have compared the results with conventional cytogenetics. Metaphase cells were available for analysis in 13 cases, of which seven showed an abnormal karyotype. CGH chromosomal profiles showed imbalances in 21 of 25 cases. The most frequent aberrations were gains of 9p (12 cases), 2q (seven cases), 3p (seven cases), chromosome 4 (seven cases), 12q (seven cases), 13q (eight cases). The main losses were at 17q and occurred in six cases. The results for CGH and cytogenetics were matched for one case only. Investigation of IMF by CGH suggests that genomic aberrations are much more common than has been previously indicated by conventional cytogenetic analysis and occur in the majority of cases. Gains of 9p were the most frequent finding, occurring in 50% of patients and suggests that genes on 9p may play a crucial role in the pathogenesis of IMF. PMID- 15801958 TI - Risks factors for highly unstable response to oral anticoagulation: a case control study. AB - The factors associated with persistent instability of oral anticoagulant treatment (OAT) were investigated in a case-control study. The most unstable patients from 35 Italian anticoagulation clinics were matched with stable controls, for gender, age and OAT indication. Socio-demographic data, medical history, dietary and life habits, cytochrome P450 CYP2C9 variants, blood cell count, liver and renal functions were investigated. An 'Abbreviated Mental Test' (AMT) and a questionnaire to assess patient compliance to, and comprehension of, OAT indications and mechanisms were administered. An International Normalized Ratio (INR) above 4.5 was more frequently found in cases (n = 77) than controls (n = 80) (12.3% vs. 0.4%; P < 0.0001). The odds ratio for instability was significantly higher for: people who worked versus pensioners, acenocoumarol versus warfarin, and an insufficient score in the AMT and/or in the questionnaire. Cytochrome P450 CYP2C9 variants *1/*3 or *2/*3 or *3/*3 were more frequent among cases than controls (29.9% vs.15.0%; P = 0.042). No differences were observed as regards the other variables. In conclusion, we found that high intra-individual variability in OAT control was multifactorial, but poor OAT comprehension was prevalent. PMID- 15801959 TI - Does cryosupernatant plasma improve outcome in thrombotic thrombocytopenic purpura? No answer yet. AB - A randomized prospective trial compared cryosupernatant plasma (CSP) to fresh frozen plasma (FFP) for treatment of thrombotic thrombocytopenic purpura (TTP). A total of 236 patients were required: 28 patients were treated with CSP and 24 with FFP within 30 months. There were no differences in survival at 1 month. By day 9, 17 of 26 patients with CSP and 18 of 24 with FFP had a platelet count >100 x 10(9)/l. At entry, von Willebrand factor (VWF) multimers were normal in all patients (range 1.1-3.95 IU/ml). ADAMTS-13 levels showed large variations ranging from 10% to 100% activity. At entry, no individual had <5% VWF cleaving protease. By day 9 (end of cycle), 89% (FFP) and 67% (CSP) had levels >50% of the controls. At 6 months some patients showed inhibitors to the enzyme in spite of adequate or normal platelet counts. The data from this study do not show an apparent advantage to the use of CSP in TTP. A large number of patients will be required to determine appropriate replacement therapy. We were not able to find a statistically significant relationship between the low level of protease activity at presentation of TTP and response. PMID- 15801960 TI - A single-centre prospective study of clinical and haemostatic risk factors for venous thromboembolism following lower limb arthroplasty. AB - Previous studies report conflicting results concerning the potential significance of thrombophilic genotypes in postarthroplasty venous thromboembolism (VTE). This study assessed thrombophilic genotypes, haemostatic and clinical variables as independent risk factors for VTE postarthroplasty. A total number of 569 patients undergoing elective lower limb arthroplasty at a single centre were prospectively studied. All patients were interviewed and had blood samples collected preoperatively. Bilateral lower limb ultrasonography was performed at day 7 +/- 2 postoperatively in all patients (ventilation/perfusion lung scanning in symptomatic patients only). The incidence of inhospital postoperative VTE was 26%. In univariate analysis - increased age, knee arthroplasty, recent surgery, general anaesthesia, shorter operation time, non-receipt of blood transfusion and differences in surgical practice (including use of pneumatic calf compression, surgical drains and postoperative bandaging techniques) were significantly associated with VTE. Factor V Leiden, prothrombin G20210A and MTHFR C677T mutations were not significant risk factors for VTE, and of all haemostatic variables tested, only median activated partial thromboplastin time showed significant difference between VTE and non-VTE patients (34 s vs. 33 s). Multiple logistic regression analysis demonstrated that increased age, knee arthroplasty and individual surgeon's routine practices were the only significant independent risks for VTE; hence routine preoperative blood screening for a potential hypercoaguable state is not indicated in this surgical setting. PMID- 15801961 TI - FRETS-VWF73, a first fluorogenic substrate for ADAMTS13 assay. AB - A plasma metalloprotease, ADAMTS13, cleaves von Willebrand factor (VWF) multimers and downregulates their activity in platelet aggregation. Functional ADAMTS13 deficiency leads to the accumulation of hyperactive large VWF multimers, inducing a life-threatening disease, thrombotic thrombocytopenic purpura (TTP). Although measuring ADAMTS13 activity is important in TTP diagnosis, existing methods require time and skill. Here, we report a fluorescence resonance energy transfer (FRET) assay for ADAMTS13 activity. We developed a synthetic 73-amino-acid peptide, FRETS-VWF73. Cleavage of this substrate between two modified residues relieves the fluorescence quenching in the intact peptide. Incubation of FRETS VWF73 with normal human plasma quantitatively increased fluorescence over time, while ADAMTS13-deficient plasma had no effect. Quantitative analysis could be achieved within a 1-h period using a 96-well format in commercial plate readers with common filters. The FRETS-VWF73 assay will be useful for the characterization of thrombotic microangiopathies like TTP and may clarify the importance of ADAMTS13 activity as a predictive marker for various thrombotic diseases. PMID- 15801962 TI - A serine-rich glycoprotein of Streptococcus sanguis mediates adhesion to platelets via GPIb. AB - Streptococcus sanguis is the most common oral bacterium causing infective endocarditis and its ability to adhere to platelets, leading to their activation and aggregation, is thought to be an important virulent factor. Previous work has shown that S. sanguis can bind directly to platelet glycoprotein (GP) Ib but the nature of the adhesin was unknown. Here, we have shown that a high molecular weight glycoprotein of S. sanguis mediates adhesion to glycocalacin. The bacterial glycoprotein was purified from cell extracts by chromatography on GPIb- and wheatgerm agglutinin affinity matrices and its interaction with GPIb was shown to be sialic acid-dependent. We designated the glycoprotein serine-rich protein A (SrpA). An insertional inactivation mutant lacking the SrpA of S. sanguis showed significantly reduced binding to glycocalacin, reduced adherence to platelets and a prolonged lag time to platelet aggregation. In addition, under flow conditions, platelets rolled and subsequently adhered on films of wild-type S. sanguis cells at low shear (50/s) but did not bind to films of the SrpA mutant. Platelets did not bind to wild-type bacterial cells at high shear (1500/s). These findings help to understand the mechanisms by which the organism might colonize platelet-fibrin vegetations. PMID- 15801963 TI - A third course of anti-thymocyte globulin in aplastic anaemia is only beneficial in previous responders. AB - This retrospective study evaluated the outcome of 18 patients with aplastic anaemia treated with a third course of anti-thymocyte globulin (ATG)-containing immunosuppressive therapy (IST). Of the 18 patients, seven had responded to one of the previous courses of ATG and 11 were refractory to both the previous courses. Self-limiting grade >/=3 liver toxicity was observed in three patients. Two patients had to discontinue ATG because of severe systemic side effects. The incidence and manifestations of serum sickness did not appear to be different during the three courses. All of the seven patients that previously responded to one of the courses responded to a third course. In contrast, of 11 patients refractory to the previous courses, only two had a transient partial response. The 3-yr event-free survival for the patients who had responded to one of the previous courses of ATG was significantly superior to that of patients refractory to both the previous courses of ATG (83% vs. 0%, P = 0.0001). For aplastic anaemia patients, a third course of ATG-containing IST is a reasonable option in previous responders. Patients refractory to previous two courses of ATG have a much lower response rate and may be suitable candidates for novel therapeutic options. PMID- 15801964 TI - Immunomodulatory effect of human adipose tissue-derived adult stem cells: comparison with bone marrow mesenchymal stem cells. AB - Like mesenchymal stem cells from bone marrow (BM-MSCs), adipose tissue-derived adult stem cells (ADAS cells) can differentiate into several lineages and present therapeutical potential for repairing damaged tissues. The use of allogenic stem cells can enlarge their therapeutical interest, provided that the grafted cells could be tolerated. We investigate here, for the first time, the immunosuppressive properties of ADAS cells compared with the well-characterized immunosuppressive properties of BM-MSCs. ADAS cells did not provoke in vitro alloreactivity of incompatible lymphocytes and, moreover, suppressed mixed lymphocyte reaction (MLR) and lymphocyte proliferative response to mitogens. The impairment of inhibition when ADAS cells and BM-MSCs were separated from lymphocytes by a permeable membrane suggests that cell contact is required for a full inhibitory effect. Hepatocyte growth factor is secreted by both stem cells but, similar to interleukin-10 and transforming growth factor-beta (TGF-beta), the levels of which were undetectable in supernatants of MLR inhibited by ADAS cells or BM-MSCs, it did not seem implicated in the stem cell suppressive effect. These findings support that ADAS cells share immunosuppressive properties with BM MSCs. Therefore, ADAS cell-based reconstructive therapy could employ allogenic cells and because of their immunosuppressive properties, ADAS cells could be an alternative source to BM-MSCs to treat allogenic conflicts. PMID- 15801965 TI - Expression of phosphatidylserine (PS) on wild-type and Gerbich variant erythrocytes following glycophorin-C (GPC) ligation. AB - Glycophorin-C (GPC) is a 40 kDa glycoprotein expressed on erythrocytes and is a receptor for the malarial parasite Plasmodium falciparum to invade these cells. A link between GPC binding (ligation) and phosphatidylserine (PS) expression on erythrocytes has been suggested by its appearance on P. falciparum-infected erythrocytes. Phosphatidylserine expression has also been shown to be a marker of cellular death in a number of biological pathways including some in erythrocytes. Using Annexin V binding, we demonstrated that ligation of GPC with mouse mAb (BRIC-10) induced PS expression on normal erythrocytes. Phosphatidylserine exposure was prevented following tryptic digestion of intact erythrocytes. In addition, GPC variant phenotypes Yus (Delta exon 2) and Gerbich (Delta exon 3), which express a truncated extracellular domain, did not express PS following BRIC 10 binding, whereas PS was exposed on Ls(a) erythrocytes (duplication of exon 3). GPC ligation was also shown to result in a concomitant loss of erythrocyte viability in wild-type erythrocytes after 24 h in vitro. These results identify a potential pathway linking GPC to PS exposure on erythrocytes that may have a role in regulating red cell turnover. Further characterization of this pathway may also identify new targets for the treatment of P. falciparum malaria. PMID- 15801966 TI - Gene expression profiling in polycythaemia vera: overexpression of transcription factor NF-E2. AB - Summary The molecular aetiology of polycythaemia vera (PV) remains unknown and the differential diagnosis between PV and secondary erythrocytosis (SE) can be challenging. Gene expression profiling can identify candidates involved in the pathophysiology of PV and generate a molecular signature to aid in diagnosis. We thus performed cDNA microarray analysis on 40 PV and 12 SE patients. Two independent data sets were obtained: using a two-step training/validation design, a set of 64 genes (class predictors) was determined, which correctly discriminated PV from SE patients. Separately 253 genes were identified to be upregulated and 391 downregulated more than 1.5-fold in PV compared with healthy controls (P < 0.01). Of the genes overexpressed in PV, 27 contained Sp1 sites: we therefore propose that altered activity of Sp1-like transcription factors may contribute to the molecular aetiology of PV. One Sp1 target, the transcription factor NF-E2 [nuclear factor (erythroid-derived 2)], is overexpressed 2- to 40 fold in PV patients. In PV bone marrow, NF-E2 is overexpressed in megakaryocytes, erythroid and granulocytic precursors. It has been shown that overexpression of NF-E2 leads to the development of erythropoietin-independent erythroid colonies and that ectopic NF-E2 expression can reprogram monocytic cells towards erythroid and megakaryocytic differentiation. Transcription factor concentration may thus control lineage commitment. We therefore propose that elevated concentrations of NF-E2 in PV patients lead to an overproduction of erythroid and, in some patients, megakaryocytic cells/platelets. In this model, the level of NF-E2 overexpression determines both the severity of erythrocytosis and the concurrent presence or absence of thrombocytosis. PMID- 15801967 TI - Alloimmunization to red blood cell antigens after universal leucodepletion. A regional multicentre retrospective study. AB - Leucodepletion has been shown to reduce human leucocyte antigen immunization, but studies on the effect of leucodepletion on red cell alloimmunization reported discordant results. We conducted a retrospective multicentre study to determine whether prestorage filter leucodepletion alters the development of clinically significant red blood cell alloimmunization against the Rhesus, Kell, Duffy, Kidd and MSs blood group systems. Two periods were investigated, 2 years before and 2 years after universal leucodepletion. Comparisons were made between the transfused patient cohorts. To control for changes not related to leucoreduction, we compared antibody incidence with antibody prevalence in the two study periods. Newly detected antibodies (n = 4770) were found in 4115 patients from 19 participating hospitals. Of these, 857 antibodies in 659 patients were because of transfusions given in the study periods. The immunization risk was 0.13% for both periods. No differences were found regarding incidence of new antibodies, nor for patients regarding age, sex, previous antibodies, multiple antibodies, additional antibodies, number of transfusions, transfusions episodes and days from transfusion to date of immunization. In conclusion, compared with buffy-coat leucoreduction, universal prestorage filter leucodepletion did not alter the development of clinically significant red blood cell alloimmunization. PMID- 15801968 TI - Criteria for defining a complete remission in acute myeloid leukaemia. PMID- 15801969 TI - Iron mobilisation from transferrin by deferiprone (L1). PMID- 15801970 TI - Amyloidosis, not myeloma. PMID- 15801973 TI - Thyrotoxic periodic paralysis after allogeneic haematopoietic stem cell transplantation. PMID- 15801975 TI - Defining the timing of respiratory syncytial virus (RSV) outbreaks: an epidemiological study. AB - BACKGROUND: Seasonal RSV infections occur every year and affect particularly children under six months of age. Passive immunoprophylaxis with monoclonal antibody Palivizumab is recommended in the period with high risk of RSV infection. This study aims to define the period for the southern part of Germany (Stuttgart area). METHODS: Epidemiological analysis of the RSV situation in southern Germany from 1996 to 2004 and comparison of results with literature was made. The respiratory tract specimens were sent in for the detection of RSV mainly by paediatric clinics. Detection of RSV was carried out mainly by real time RT-PCR or by ELISA "Pathfinder". RSV outbreaks were depicted as an absolute number and as a percentage of RSV diagnoses in a month. Onsets, offsets, peaks, duration and severity of RSV seasons were defined and analysed. RESULTS: An early season with strong RSV activity (early-high phase) was followed by a weaker late season (late-low phase) in a regular biennial rhythm. However, onsets, offsets and durations of outbreaks varied significantly from year to year. RSV epidemics in southern Germany were found to oscillate in an antiphase with RSV epidemics in Finland and Sweden. CONCLUSION: The long-term regular biennial rhythm allows predicting whether the next outbreak will be late or early and whether RSV activity will be strong or weak. Not foreseeable, however, is the precise time of increase and decrease of RSV activity. Moreover, the regular seasonal pattern may be disrupted by irregular outbreaks. Thus, activity of RSV has to be monitored every year to define the period with high risk of infection. PMID- 15801976 TI - Development of daily rhythmicity in heart rate and locomotor activity in the human fetus. AB - BACKGROUND: Very little is known about the perinatal genesis of circadian rhythmicity in the human fetus. Some researchers have found evidence of rhythmicity early on in fetal development, whereas others have observed a slow development of rhythmicity during several years after birth. METHOD: Rhythms of fetal heartbeat and locomotor activity were studied in women with physiological course of pregnancy at 16 to 40 gestational weeks. Observations were conducted continuously for 24 h using the method of external electrocardiography, which provided simultaneous detection of the changes in maternal and fetal heartbeat as well as assessment of daily locomotor activity of the fetus. During the night time, electroencephalogram, myogram, oculogram and respiration of the mother were registered in parallel with fetal external electrocardiography. RESULTS: Although we found no significant daily rhythmicity in heart rate per se in the human fetus, we developed a new method for the assessment of 24-h fetal cardiotachogram that allowed us to identify daily rhythmicity in the short-term pattern of heart beating. We found that daily rhythmicity of fetal electrocardiogram resembles that of the mother; however, the phase of the rhythm is opposite to that of the mother. "Active" (from 9 a.m. to 2 p.m. and from 7 p.m. to 4 a.m.) and "quiet" (from 4 a.m. to 9 a.m. and from 2 p.m. to 7 p.m.) periods of activity were identified. CONCLUSION: A healthy fetus at gestational age of 16 to 20 weeks reveals pronounced rhythms of activity and locomotion. Absence of distinct rhythmicity within the term of 20 to 24 weeks points to developmental retardation. The "Z"-type fetal reaction, recorded during the "quiet" hours, does not indicate unsatisfactory state, but rather is suggestive of definite reduction of functional levels of the fetal physiological systems necessary for vital activity. PMID- 15801977 TI - The optimal starting time of postoperative intraperitoneal mitomycin-C therapy with preserved intestinal wound healing. AB - BACKGROUND: There is controversy about the effect of the timing of intraperitoneal administration of chemotherapeutic agents on the healing of intestinal anastomosis. We have investigated the effect on intestinal wound healing of mitomycin-C administered at different times post-operatively. METHODS: Eighty-four Wistar-Albino female rats underwent ileal resection and end-to-end anastomosis. The rats were randomly selected for intraperitoneal administration of mitomycin-C or saline as follows: mitomycin-C group (n = 65), 2 mg/kg mitomycin-C; control group (n = 13), 10 ml saline. The former was sub-divided into 5 equal groups (A 1-5) and mitomycin-C was administered postoperatively as follows: day 0 (A1), day 3 (A2), day 5 (A3), day 7 (A4) and day 10 (A5). All the rats were sacrificed on the 14th postoperative day and anastomotic bursting pressures and tissue hydroxyproline levels were determined. RESULTS: Five of the animals died postoperatively: 2 (15.4%) in group A1, 2 (15.4%) in group A2 and 1(7.7%) in group A3. Non-lethal anastomotic leakage was observed in a further five animals: 1 in group A1, 2 in group A2, 1 in group A5 and 1 in the control group. Groups A1 and A2 had significantly lower anastomotic bursting pressures than the other groups (P was <0.05 for each comparison). The anastomotic bursting pressures of group A3, A4 and A5 were comparable with those of the controls (P was >0.05 for each comparison). Tissue hydroxyproline levels in group A1 and A2 were significantly lower than in the controls (P values were <0.05 for each comparison) or the other mitomycin-C sub-groups (P was <0.05 for each comparison). CONCLUSIONS: Intraperitoneal chemotherapy impairs intestinal wound healing when applied before the 5th postoperative day. Additional therapeutic approaches are needed to prevent this potentially lethal side effect of early intraperitoneal mitomycin-C administration. PMID- 15801978 TI - Epidermal growth factor mediates detachment from and invasion through collagen I and Matrigel in Capan-1 pancreatic cancer cells. AB - BACKGROUND: Pancreatic adenocarcinoma is a highly invasive neoplasm. Epidermal growth factor (EGF) and its receptor are over expressed in pancreatic cancer, and expression correlates with invasion and metastasis. We hypothesized that EGF receptor and integrin signalling pathways interact in mediating cellular adhesion and invasion in pancreatic cancer, and that invasiveness correlates temporally with detachment from extracellular matrix. METHODS: We tested this hypothesis by investigating the role of EGF in mediating adhesion to and invasion through collagen I and Matrigel in the metastatic pancreatic adenocarcinoma cell line Capan-1. Adhesion and invasion were measured using in vitro assays of fluorescently-labeled cells. Adhesion and invasion assays were also performed in the primary pancreatic adenocarcinoma cell line MIA PaCa-2. RESULTS: EGF inhibited adhesion to collagen I and Matrigel in Capan-1 cells. The loss of adhesion was reversed by AG825, an inhibitor of erbB2 receptor signalling and by wortmannin, a PI3K inhibitor, but not by the protein synthesis inhibitor cycloheximide. EGF stimulated invasion through collagen I and Matrigel at concentrations and time courses similar to those mediating detachment from these extracellular matrix components. Adhesion to collagen I was different in MIA PaCa 2 cells, with no significant change elicited following EGF treatment, whereas treatment with the EGF family member heregulin-alpha elicited a marked increase in adhesion. Invasion through Matrigel in response to EGF, however, was similar to that observed in Capan-1 cells. CONCLUSION: An inverse relationship exists between adhesion and invasion capabilities in Capan-1 cells but not in MIA PaCa-2 cells. EGF receptor signalling involving the erbB2 and PI3K pathways plays a role in mediating these events in Capan-1 cells. PMID- 15801979 TI - Columba: an integrated database of proteins, structures, and annotations. AB - BACKGROUND: Structural and functional research often requires the computation of sets of protein structures based on certain properties of the proteins, such as sequence features, fold classification, or functional annotation. Compiling such sets using current web resources is tedious because the necessary data are spread over many different databases. To facilitate this task, we have created COLUMBA, an integrated database of annotations of protein structures. DESCRIPTION: COLUMBA currently integrates twelve different databases, including PDB, KEGG, Swiss-Prot, CATH, SCOP, the Gene Ontology, and ENZYME. The database can be searched using either keyword search or data source-specific web forms. Users can thus quickly select and download PDB entries that, for instance, participate in a particular pathway, are classified as containing a certain CATH architecture, are annotated as having a certain molecular function in the Gene Ontology, and whose structures have a resolution under a defined threshold. The results of queries are provided in both machine-readable extensible markup language and human-readable format. The structures themselves can be viewed interactively on the web. CONCLUSION: The COLUMBA database facilitates the creation of protein structure data sets for many structure-based studies. It allows to combine queries on a number of structure related databases not covered by other projects at present. Thus, information on both many and few protein structures can be used efficiently. The web interface for COLUMBA is available at http://www.columba-db.de. PMID- 15801980 TI - Increase in number of circulating disseminated epithelial cells after surgery for non-small cell lung cancer monitored by MAINTRAC(R) is a predictor for relapse: A preliminary report. AB - BACKGROUND: Lung cancer still remains one of the most commonly occurring solid tumors and even in stage Ia, surgery fails in 30% of patients who develop distant metastases. It is hypothesized that these must have developed from occult circulating tumor cells present at the time of surgery, or before. The aim of the present study was to detect such cells in the peripheral blood and to monitor these cells following surgery. METHODS: 30 patients treated for lung cancer with surgery were monitored for circulating epithelial cells (CEC) by taking peripheral blood samples before, 2 weeks and 5 months after surgery and/or radiotherapy (RT) chemotherapy (CT) or combined RT/CT using magnetic bead enrichment and laser scanning cytometry (MAINTRAC(R)) for quantification of these cells. RESULTS: In 86% of the patients CEC were detected before surgery and in 100% at 2 weeks and 5 months after surgery. In the control group, which consisted of 100 normal donors without cancer, 97 % were negative for CEC. A significantly higher number of CEC was found preoperatively in patients with squamous cell carcinoma than in those with adenocarcinoma. In correlation to the extent of parenchymal manipulation 2 weeks after surgery, an increase in numbers of CEC was observed with limited resections (18/21) whereas pneumonectomy led to a decrease (5/8) of CEC, 2 weeks after surgery. The third analysis done 5 months after surgery identified 3 groups of patients. In the group of 5 patients who received neo- or adjuvant chemo/radiotherapy there was evidence that monitoring of CEC can evaluate the effects of therapy. Another group of 7 patients who underwent surgery only showed a decrease of CEC and no signs of relapse. A third group of 11 patients who had surgery only, showed an increase of CEC (4 with an initial decrease after surgery and 7 with continuous increase). In the group with a continuous increase during the following 24 months, 2 early relapses in patients with stage Ia adenocarcinoma were observed. The increase of CEC preceded clinical detection by six months. CONCLUSION: We consider, therefore, that patients with adenocarcinoma and a continuous increase of CEC after complete resection for lung cancer are at an increased risk of early relapse. PMID- 15801982 TI - Buying best value health care: Evolution of purchasing among Australian private health insurers. AB - Since 1995 Australian health insurers have been able to purchase health services pro-actively through negotiating contracts with hospitals, but little is known about their experience of purchasing. This paper examines the current status of purchasing through interviews with senior managers representing all Australian private health insurers. Many of the traditional tools used to generate competition and enhance efficiency (such as selective contracting and co payments) have had limited use due to public and political opposition. Adoption of bundled case payment models using diagnosis related groups (DRGs) has been slow. Insurers cite multiple reasons including poor understanding of private hospital costs, unfamiliarity with DRGs, resistance from the medical profession and concerns about premature discharge. Innovation in payment models has been limited, although some insurers are considering introduction of volume-outcome purchasing and pay for performance incentives. Private health insurers also face a complex web of regulation, some of which appears to impede moves towards more efficient purchasing. PMID- 15801981 TI - Scale and shape issues in focused cluster power for count data. AB - BACKGROUND: Interest in the development of statistical methods for disease cluster detection has experienced rapid growth in recent years. Evaluations of statistical power provide important information for the selection of an appropriate statistical method in environmentally-related disease cluster investigations. Published power evaluations have not yet addressed the use of models for focused cluster detection and have not fully investigated the issues of disease cluster scale and shape. As meteorological and other factors can impact the dispersion of environmental toxicants, it follows that environmental exposures and associated diseases can be dispersed in a variety of spatial patterns. This study simulates disease clusters in a variety of shapes and scales around a centrally located single pollution source. We evaluate the power of a range of focused cluster tests and generalized linear models to detect these various cluster shapes and scales for count data. RESULTS: In general, the power of hypothesis tests and models to detect focused clusters improved when the test or model included parameters specific to the shape of cluster being examined (i.e. inclusion of a function for direction improved power of models to detect clustering with an angular effect). However, power to detect clusters where the risk peaked and then declined was limited. CONCLUSION: Findings from this investigation show sizeable changes in power according to the scale and shape of the cluster and the test or model applied. These findings demonstrate the importance of selecting a test or model with functions appropriate to detect the spatial pattern of the disease cluster. PMID- 15801983 TI - Ultrastructural studies on dengue virus type 2 infection of cultured human monocytes. AB - BACKGROUND: Early interaction of dengue virus and monocyte/macrophages could be an important feature for virus dissemination after its initial entry via the mosquito vector. Since ultrastructural analysis of this interaction has not been reported, dengue type 2 (DEN2) virus-infected human monocyte cultures were studied at 1, 2, 4 and 6 hours after infection. RESULTS: Typical dengue particles and fuzzy coated viral particles were 35 to 42 nm and 74 to 85 nm respectively. Viruses were engulfed by phagocytosis and macropicnocytosis leading to huge vacuoles and phagosomes inside the monocytes. Interaction of monocytes with DEN2 virus induced apoptosis, characterized by nuclear condensation and fragmentation, cellular shrinkage, blebbing and budding phenomena and phagocytosis of apoptotic cells by neighboring monocytes. This finding was confirmed by TUNEL. Ultrastructural features associated to DEN2 virus replication were not observed. CONCLUSION: These data suggest that clearance of the virus by monocytes and cellular death are the main features during the initial interaction of DEN2 virus and monocytes and this could be important in the rapid elimination of the virus after infection by mosquito vector. PMID- 15801984 TI - Public health education: a report from Mosul and a plan for change. AB - BACKGROUND: Today Iraq suffers from severe shortages of food, medicine, clean water and adequate sanitation. Malnutrition and communicable diseases are major factors in the rising morbidity and mortality rates. However, supplies and equipment are insufficient or outmoded, and public health training is outdated. The Universities have been unable to help because under-funding and isolation from their professional colleagues has limited their effectiveness. METHODS: To revitalize public health education, we describe a partnership between a US education consortium and the University of Mosul that will be carried out in the next several years. The plan is based on "three R's": Recovery from the past damage due to war and neglect; Retooling of key public health faculty to remedy the years of isolation and restriction of activity; and Reestablishment of the University as a resource for the its constituents, for the community and for other educational institutions. In all these activities, Iraqi minorities, especially women, will participate and contribute. CONCLUSION: The work to repair the public health educational infrastructure has just begun. When completed, it will represent a small but necessary step in restoring normalcy to the people of Mosul, and of Iraq. PMID- 15801985 TI - General practitioners' satisfaction with and attitudes to out-of-hours services. AB - BACKGROUND: In recent years, Dutch general practitioner (GP) out-of-hours service has been reorganised into large-scale GP cooperatives. Until now little is known about GPs' experiences with working at these cooperatives for out-of-hours care. The purpose of this study is to gain insight into GPs' satisfaction with working at GP cooperatives for out-of-hours care in separated and integrated cooperatives. METHODS: A GP cooperative separate from the hospital Accident and Emergency (A&E) department, and a GP cooperative integrated within the A&E department of another hospital. Both cooperatives are situated in adjacent geographic regions in the South of The Netherlands. One hundred GPs were interviewed by telephone; fifty GPs working at the separated GP cooperative and fifty GPs from the integrated GP cooperative. Opinions on different aspects of GP cooperatives for out-of-hours care were measured, and regression analysis was performed to investigate if these could be related to GP satisfaction with out-of hours care organisation. RESULTS: GPs from the separated model were more satisfied with the organisation of out-of-hours care than GPs from the integrated model (70 vs. 60 on a scale score from 0 to 100; P = 0.020). Satisfaction about out-of-hours care organisation was related to opinions on workload, guarantee of gatekeeper function, and attitude towards out-of-hours care as being an essential part of general practice. Cooperation with medical specialists was much more appreciated at the integrated model (77 vs. 48; P < 0.001) versus the separated model. CONCLUSION: GPs in this study appear to be generally satisfied with the organisation of GP cooperatives for out-of-hours care. Furthermore, GPs working at the separated cooperative seem to be more satisfied compared to GPs working at the integrated cooperative. PMID- 15801986 TI - Downregulation of calcineurin activity in cervical carcinoma. AB - BACKGROUND: Calcineurin (CaN) is an important serine-threonine phosphatase (PP2B), which plays a crucial role in calcium-calmodulin mediated signal transduction events. Calcineurin has been implicated in pathogenesis of various diseases cardiac hypertrophy, diabetic neuropathy and Alzheimer's, however its role in neoplasia remains unclear. RESULTS: In view of this we evaluated the calcineurin activity in serum and biopsy samples collected from women diagnosed with invasive squamous cell carcinoma of cervix. A significant reduction was observed in the calcineurin activity in cancer cervix patients compared to the control group. However the calcineurin activity remained unaltered in the cervical scrapes obtained from patients diagnosed with low-grade squamous intra epithelial lesions (LSIL). Interestingly the downregulation of calcineurin activity in squamous cell carcinomas was not accompanied by any significant change in DNA-binding affinity of the transcriptional factor NFAT (Nuclear Factor of Activated T-cells). All the squamous cell carcinoma samples used in the present study were positive for high-risk human papillomavirus (HPV) types. CONCLUSION: The present study demonstrates the downregulation of calcineurin activity in squamous cell carcinoma of cervix with high risk HPV infection. We conclude that perturbations in calcineurin-mediated pathway may be involved in development of cervical neoplasia. PMID- 15801987 TI - Patients with obsessive-compulsive disorder have increased 5-HT2A receptor binding in the caudate nuclei. AB - The pharmacological efficacy of serotonergic-acting drugs suggest that patients with obsessive-compulsive disorder (OCD) may have alterations in their cerebral serotonergic (5-HT) receptor system, and previous neuroimaging studies of OCD patients have shown abnormalities in several fronto-subcortical regions. In this study we investigated cerebral 5-HT(2A) receptor binding in 15 untreated OCD patients and in 15 age- and gender-matched healthy volunteers by magnetic resonance imaging and [(18)F]altanserin positron emission tomography (PET). Eleven of the patients were rescanned with PET after receiving treatment with a selective serotonin reuptake inhibitor (SSRI). The distribution volumes of specific tracer binding (DV(3)') were calculated for 12 brain regions, and comparisons were made between: (1) healthy volunteers vs. untreated OCD patients, (2) healthy volunteers vs. treated OCD patients, and (3) OCD patients before and during treatment. When comparing the distribution volume for specific fronto subcortical brain regions, significantly higher values were recorded in the caudate nuclei in OCD patients (DV(3)': 0.24+/-0.14) compared to the healthy control group (DV(3)': 0.15+/-0.13) (p<0.05, Wilcoxon matched-pairs test). This difference between groups was not present after treatment with SSRIs. There was no correlation between the severity of OCD symptoms and 5-HT(2A )receptor binding. An increase in 5-HT(2A) receptor binding is found in the caudate nuclei of untreated patients with OCD. The up-regulation in 5-HT(2A) receptors might be compensatory for a lack of serotonin in the feedback loop between the thalamus and orbito-frontal cortex, the caudate nuclei, and the globus pallidus. PMID- 15801988 TI - Self-monitoring of blood glucose as part of a multi-component therapy among non insulin requiring type 2 diabetes patients: a meta-analysis (1966-2004). AB - OBJECTIVE: To determine if therapeutic management programs that include self monitoring of blood glucose result in greater HbA1c reduction in non-insulin requiring type 2 diabetes patients compared to programs without blood glucose self-monitoring. RESEARCH DESIGN AND METHODS: Electronic databases including MEDLINE (1966-2004), Cochrane Database of Systematic Reviews, EMBASE (1950-2004), Centre for Reviews and Dissemination (CRD) and the Online Index Journals of the American Diabetes Association (ADA 1978-2004) were searched. Personal collections of investigators were also explored. Randomized controlled trials comparing HbA1c reduction in therapies with and without blood glucose self-monitoring among adult, non-insulin-treated type 2 diabetes patients were selected. Studies on patients who are pregnant, taking insulin, troglitazone or experimental drugs were excluded. Out of 14 potentially useful randomized controlled trials on self monitoring of blood glucose in non-insulin treated type 2 diabetes patients, eight studies with a total of 1307 subjects were included in the analysis. Two independent reviewers assessed the quality of studies. MAIN OUTCOME MEASURE: The effect of SMBG was assessed by means of meta-analysis of the difference in HbA1c reduction between self-monitoring and non-self-monitoring groups. RESULTS: Antidiabetic therapies that included blood glucose self-monitoring as part of a multi-component management strategy produced a mean additional HbA(1c) reduction of -0.39% (95%CI: -0.54%, -0.23%) under the fixed effects model and -0.42% (95%CI: -0.63%, -0.21%) under the random effects model, when compared to therapies that did not. Heterogeneity among studies was not statistically significant. CONCLUSION: Multi-component diabetes management programs with self monitoring of blood glucose result in better glycemic control among non-insulin using type 2 diabetes patients. PMID- 15801989 TI - Trabecular bone microarchitecture after alendronate treatment of osteoporotic women. AB - OBJECTIVE: To compare the microarchitecture of iliac crest trabecular bone from women treated for two to three years with alendronate versus that of women treated with placebo. RESEARCH DESIGN AND METHODS: Three-dimensional micro computed tomography (micro-CT; resolution 20 microm) and two-dimensional histomorphometry (resolution 5-7 microm) were used to examine trabecular bone from single transilial biopsies obtained at the completion of clinical trials. MAIN OUTCOME MEASURES: Microarchitectural variables, including bone volume, trabecular number, trabecular thickness, and trabecular spacing in specimens from alendronate- and placebo-treated women were examined. Three-dimensional images of trabecular bone from both groups were constructed from CT images. Correlations among variables and between techniques were also calculated. RESULTS: Eighty eight specimens were suitable for evaluation by both techniques. As measured by two-dimensional histomorphometry, bone volume fraction (as a proportion of total volume) and trabecular thickness were significantly greater in alendronate specimens, 17.1 +/- 5.5% vs. 13.4 +/- 5.5% (p = 0.0043) and 127 +/- 29 microm vs. 109 +/- 28 microm (p = 0.0090), respectively, and trabecular spacing was significantly smaller, 729 +/- 227 microm vs. 862 +/- 338 microm (p = 0.005). Micro-CT yielded similar findings: bone volume and trabecular number were significantly greater in alendronate specimens: 19.4 +/- 6.2% vs. 16.2 +/- 6.3% (p = 0.0412) and 1.46(+/-) 0.32 vs. 1.31(+/-) 0.33 per mm (p = 0.0346). Two dimensional and micro-CT measured characteristics correlated strongly with one another, with Pearson product moment correlation coefficients ranging from 0.60 (for trabecular thickness) to 0.83 (for bone volume). CONCLUSIONS: Trabecular microarchitecture of the ilium, whether studied by two- or three-dimensional methods, is better (greater bone volume, greater trabecular thickness, decreased trabecular spacing) after alendronate treatment than after two to three years of treatment with placebo. Bone volume in a trabecular region is strongly correlated to its microarchitecture, suggesting that bone quantity predicts values for these microarchitectural endpoints. PMID- 15801990 TI - Costs of attention deficit-hyperactivity disorder (ADHD) in the US: excess costs of persons with ADHD and their family members in 2000. AB - OBJECTIVE: The objective of this study is to provide a comprehensive estimate of the cost of ADHD by consider ing the healthcare and work loss costs of persons with ADHD, as well as those costs imposed on their family members. METHODS: Excess per capita healthcare (medical and prescription drug) and work loss (disability and work absence) costs of treated ADHD patients (ages 7 years-44 years) and their family members (under 65 years of age) were calculated using administrative claims data from a single large company; work loss costs are from disability data or imputed for medically related work loss days. Excess costs are the additional costs of patients and their family members over and above those of comparable control individuals. The excess costs of untreated individuals with ADHD and their family members were also estimated. All per capita costs were extrapolated using published prevalence and treatment rates and population data; the prevalence of persons with ADHD was based upon the literature. RESULTS: The total excess cost of ADHD in the US in 2000 was $31.6 billion. Of this total, $1.6 billion was for the ADHD treatment of patients, $12.1 billion was for all other healthcare costs of persons with ADHD, $14.2 billion was for all other healthcare costs of family members of persons with ADHD, and $3.7 billion was for the work loss cost of adults with ADHD and adult family members of persons with ADHD. CONCLUSION: The annual cost of ADHD in the US is substantial. Both treated and untreated persons with ADHD, as well as their family members, impose consider able economic burdens on the healthcare system as a result of this condition. While these first estimates of the cost of ADHD to the nation are suggestive of its substantial economic burden, future research needs to refine and build on this analysis, particularly in the context of a model to control for related co morbidities. Similarly, since these results are based on data from a single company for the period 1996-1998, the analysis should be validated with more representative, current data. PMID- 15801991 TI - Effects of switching pravastatin to cerivastatin on C-reactive protein, butyrylcholinesterase, and lipids. AB - OBJECTIVE: C-reactive protein (CRP) concentrations, butyrylcholinesterase (BChE) activity, total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglycerides (TG) were evaluated in patients switched from pravastatin to cerivastatin. The purpose of this study was to determine whether a more potent statin (cerivastatin) would further affect CRP, whether a relation ship between CRP and BChE existed, and if there were any relationships between CRP or BChE and lipids. In view of the withdrawal of cerivastatin from the market, studies considering the effects of conversion of patients from one statin to another are warranted. RESEARCH DESIGN AND METHODS: Thirty-seven patients actively taking pravastatin (10 mg-40 mg) were switched to cerivastatin (0.2 mg-0.8 mg) at the initial visit in the Lipid Clinic at David Grant Medical Center, Travis Air Force Base. Samples were collected before the conversion (pravastatin phase) and at 6 weeks and 12 weeks post-conversion. Patients were excluded from the study if they were taking gemfibrozil concomitantly. Patients were counseled on the adverse effects of cerivastatin, including rhabdomyolsis. RESULTS: Median CRP levels at the pravastatin phase, 6 weeks of cerivastatin, and 12 weeks of cerivastatin, were 0.380 mg/dL, 0.403 mg/dL, and 0.364 mg/dL (p = 0.772), respectively. Median BChE activity at the pravastatin phase, 6 weeks of cerivastatin, and 12 weeks of cerivastatin were 0.338 micromol/mL/min, 0.332 micromol/mL/min, 0.33 micromol/mL/min (p = 0.746), respectively. A negative correlation was observed between CRP and BChE at baseline only (r = -0.353, p = 0.032). There was a significant decline in mean TC (p < 0.001) and median LDL (p < 0.001) and a significant increase in mean HDL (p = 0.017) over the three time points. Numerically TG declined, but it was not statistically significant (p = 0.649). No correlations were observed between CRP or BChE and any of the lipids. Gender, aspirin use, and the presence of CHD or diabetes did not affect CRP levels or BChE activity. CONCLUSION: Median CRP remained stable with pravastatin and cerivastatin use, although TC and LDL decreased. The further decline observed with LDL, but not CRP suggests differing effects of statins on LDL and CRP. Limitations include no serum levels prior to statin use and small sample size; thus, future studies are needed to address the relationship between cholesterol and CRP and the mechanism of action of statins on CRP. PMID- 15801992 TI - Anticoagulation therapy in patients with chronic atrial fibrillation: a retrospective claims data analysis. AB - OBJECTIVES: This study assessed the risk of thrombo embolic events and bleeding complications among atrial fibrillation patients. METHODS: A cohort of patients with chronic non-valvular atrial fibrillation were identified from medical claims (diagnosis codes 427.31 and 427.32). Subjects were identified from 1 January 1998 31 December 2000 and were continuously enrolled for 6 months prior to the first occurring atrial fibrillation medical claim. Cox proportional hazards analysis with time varying covariates was used for the event analysis. RESULTS: Of 6764 subjects retained for analysis, 3541 (52.4%) were exposed to warfarin. Adjusting for baseline characteristics, warfarin exposure was associated with lower likelihood of an arterial thromboembolic event compared to no exposure (HR: 0.710, CI: 0.540-0.934). No benefit was found in the use of warfarin in the prevention of intracranial events (HR: 1.119, CI: 0.929-1.349). Use of warfarin increased the risk of minor bleeding events (HR: 3.600, CI: 2.537-5.109), and all bleeding events (HR: 1.502, CI: 1.289-1.749). CONCLUSIONS: The risk of arterial thromboembolic events was associated with warfarin exposure as expected. An increase in the risk of minor and total bleeding events among patients treated with warfarin was observed. The results of this study suggest that there may be a gap between the clinical trial and coagulation clinic performance of warfarin in reducing the risk of thromboembolic events versus what is achievable in general practice. PMID- 15801993 TI - Effects of temazepam 7.5 mg and temazepam 15 mg on sleep maintenance and sleep architecture in a model of transient insomnia. AB - OBJECTIVE: To compare the effects of temazepam 7.5 mg and temazepam 15 mg on sleep maintenance during the last third of the night (last 160 min) and on sleep architecture throughout the night. RESEARCH DESIGN AND METHODS: This was a retrospective analysis of a previously reported double-blind, randomized, uncontrolled, parallel-group, multicenter study. Healthy subjects with previous but no current complaints of transient insomnia were enrolled. Transient insomnia was induced in the sleep laboratory by means of the 'first night' effect and by implementing a 2-h phase advance. The effects of both doses of temazepam on polysomnographic measures of sleep were evaluated for 1 night. The primary, prospectively-defined analysis of this study showed that 7.5-mg and 15-mg doses of temazepam had equivalent effects on latency to persistent sleep, total sleep time, and the number of sleep interruptions recorded over an 8-h period. Both doses were well tolerated. The post hoc analysis reported here compared these 2 doses for their effects on sleep maintenance and architecture. Sleep efficiency during the last third of the night was designated as the primary endpoint. The methodology for this analysis was fully defined and documented prior to re analysis of the database for these parameters. RESULTS: Sixty-five subjects received temazepam 7.5 mg and 66 received temazepam 15 mg. No statistically significant differences between doses were detected for sleep efficiency or number of sleep interruptions during the last third of the night. Sleep architecture (measured over 8 h) did not differ significantly between groups. CONCLUSIONS: The 7.5-mg and 15-mg doses of temazepam were equally effective for maintaining sleep during the last third of the night. Continuity of sleep throughout the night, as reflected by sleep architecture, was also similar regardless of dose. In keeping with current practice guidelines, initiation of treatment with temazepam for transient insomnia should begin with the 7.5 mg dose. PMID- 15801994 TI - A review of the lipid-related effects of fluvastatin. AB - BACKGROUND: Statin therapy has been shown to significantly decrease vascular events and overall mortality in primary and secondary prevention trials. This review considers the pharmacology, nonlipid-lowering effects and clinical trial evidence of fluvastatin based on a survey of PubMed entries. FINDINGS: Recent clinical data show that treatment with fluvastatin is associated with a variety of benefits in different high-risk populations along with a good safety profile. Fluvastatin exerts non-lipid lowering-associated pleiotropic effects in both clinical and experimental studies. Furthermore, an extended-release formulation of fluvastatin with a favourable pharmacokinetic profile is available. CONCLUSION: Treatment with fluvastatin offers a convenient, safe and evidence based approach to managing dyslipidaemias and preventing vascular events. PMID- 15801995 TI - Comparative efficacy of once daily, 5-day short-course therapy with clarithromycin extended-release versus twice daily, 7-day therapy with clarithromycin immediate-release in acute bacterial exacerbation of chronic bronchitis. AB - OBJECTIVE: The objective of this study was to compare the efficacy, tolerability, and safety of two clarithromycin regimens, extended-release (ER) 1000 mg once daily for 5 days and immediate-release (IR) 500 mg twice daily for 7 days, in the treatment of acute bacterial exacerbation of chronic bronchitis (ABECB). PATIENTS AND METHODS: This was a double-blind, randomized, parallel-group, multicenter study of ambulatory patients at least 40 years old with a presumptive diagnosis of ABECB, purulent sputum, and documented evidence of chronic obstructive pulmonary disease (COPD), including forced expiratory volume in one second (FEV(1)) < 70% of predicted value. Clinical cure, bacteriological cure, and target pathogen eradication rates were determined at a test-of-cure visit (study days 14-40). Safety was assessed based on the incidence of study drug-related adverse events. RESULTS: A total of 485 patients were randomized (240 to ER and 245 to IR). Clinical cure rates were similar for evaluable patients treated with ER (84%, 157/187) and those treated with IR (84%, 172/204) (95% CI -7.9, 7.2). The bacteriological cure rates were 87% (82/94) and 89% (91/102), and the overall target pathogen eradication rates were 88% (107/122) and 89% (117/131) for the respective treatment groups. The incidence of adverse events was 13% (31/240) in the ER group and 18% (45/245) in the IR group. The rate of gastrointestinal adverse events was lower with ER (8%, 19/240) compared to IR (11%, 26/245). Clarithromycin ER-treated patients reported statistically significantly fewer adverse events due to abnormal taste than did clarithromycin IR-treated patients (3% and 8%, respectively, p = 0.012). CONCLUSION: Both once-daily, 5-day, short course therapy with clarithromycin ER and 7-day, twice-daily therapy with clarithromycin IR were effective in resolving clinical signs/symptoms of ABECB and eradicating the causative pathogens, with no statistically significant difference in clinical cure rate between the treatment groups. Clarithromycin ER was better tolerated, causing fewer gastrointestinal adverse events and statistically significantly fewer reports of abnormal taste as compared with clarithromycin IR. PMID- 15801996 TI - An open-label evaluation of HP-Guar gellable lubricant eye drops for the improvement of dry eye signs and symptoms in a moderate dry eye adult population. AB - OBJECTIVE: Evaluate the efficacy of a polymer hydroxypropyl guar (HP-Guar) gellable lubricant eye drop (Systane* Lubricant Eye Drops) in reducing dry eye signs and symptoms among dry eye patients who exhibited at least moderate signs and symptoms. METHODS: 168 patients with moderate dry eye signs and symptoms were enrolled at 29 sites in this open label study. The mean age of patients was 62 years with a minimum age of 28 years and a maximum of 90 years. One hundred and forty-seven patients completed the study, 111 female and 35 male, excluding 1 subject (gender not captured). In order to be included in the study, subjects were required to have a total corneal staining score > or = 4 (NEI grid) in at least one eye, with a grade > or = 2 in at least one zone of the same eye. Patients also had to indicate that their eyes 'felt dry enough to want to use eye drops' at least 'some of the time' on a standardized frequency scale. Eligible patients were dispensed a run-in drop (Opti-Free Express Rewetting Drops*) to use QID for 7 days, and then examined. Patients continuing to meet the inclusion criteria were dispensed the test drops (HP-Guar gellable lubricant eye drops) to use QID, and re-examined on Day 28. At each visit, corneal and conjunctival staining were measured, and six ocular discomfort symptoms were rated on a standardized 0-4 severity scale. At Days 0 and 28, patients subjectively rated product acceptability using a Likert scale. RESULTS: No significant changes in corneal or conjunctival staining were observed with the use of the run-in drop. After 28 days of test drop use, there was a statistically significant reduction in corneal staining (p < 0.0001). Ninety-four percent of patients improved from baseline, with mean reduction in total corneal staining of 4.1 units (0-15 total scale) (62%). Conjunctival staining also improved significantly (p < 0.0001) with a mean total reduction of 3.1 (59%). Patients experienced statistically significant symptomatic relief from day 0 to day 28 for all six ocular discomfort severity questions (p < 0.0001). CONCLUSION: Lubricating drops effectively relieved signs and symptoms associated with moderate dry eye, with measurable improvements evident in both objective staining and subjective questionnaire measures after 28 days in this study population. PMID- 15801997 TI - An economic and outcomes assessment of first-line monotherapy in the treatment of community-acquired pneumonia within managed care. AB - OBJECTIVE: To evaluate the resource consumption and outcomes associated with first-line monotherapy for community-acquired pneumonia, focusing specifically on the use of erythromycin, azithromycin, clarithromycin, and levofloxacin. STUDY DESIGN: Retrospective managed care database analysis. PATIENTS AND METHODS: Subjects included patients within a managed care setting over 18 years of age with an initial diagnosis of community-acquired pneumonia from January 1995 to April 2002. Multivariate linear and logistic regression models were used to examine associations with treatment success rates and direct medical costs between antibiotic treatments after controlling for patient demographics and pneumonia risk factors. MAIN RESULTS: Overall, treatment success rates were high (95.8%), the use of second antibiotics was un common (2.3%), and hospitalizations were infrequent (2.0%) among the 1952 subjects studied. After controlling for patient characteristics and risk factors, significantly lower total costs were associated with erythromycin (92.7% lower, p < 0.001), azithromycin (48.7% lower, p < 0.001), and clarithromycin (21.3% lower, p = 0.015) relative to levofloxacin, with no difference in treatment success between groups. Among newer agents, azithromycin (49.2% lower, p < 0.001) and clarithromycin (21.7% lower, p = 0.013) treatment groups were associated with significantly lower total costs than levofloxacin in the full sample. However, in subjects with a chronic disease score above the sample's mean, only azithromycin was associated with significantly lower total costs (47.9% lower, p < 0.001) relative to levofloxacin. CONCLUSION: Erythromycin, azithromycin, and clarithromycin were associated with significantly lower total costs than levofloxacin, although treatment success rates did not differ between groups. Following stratification based upon various subset criteria, erythromycin and azithromycin were observed to have significantly lower total costs than levofloxacin. Although these findings may augment clinical guidelines and evidence-based approaches, health plans should consider evaluating their own patient data to see if similar differences exist in their populations. PMID- 15801998 TI - A comparison of the resources used in advanced cancer care between two different strong opioids: an analysis of naturalistic practice in the UK. AB - OBJECTIVE: To assess the resource implications of using strong opioids in patients with advanced cancer in the UK, based on naturalistic practice, in order to develop the evidence base supporting better management. DESIGN AND SETTING: A modelling study performed from the perspective of the UK's National Health Service (NHS). Study participants and interventions: A data set was created from the DIN-link database comprising 986 patients with advanced cancer who were prescribed either 12-hourly sustained release morphine (SR morphine; MST Continuous) ( n = 784) or transdermal fentanyl (Durogesic) (n = 202) as their first strong opioid between 1st January 1998 and 30th September 2000 and died during that period. METHODS: Palliative care-related resource use data were obtained from the DIN-link database. Unit costs at 2000/2001 prices were applied to the resource use values to determine the mean NHS cost of palliative care from the start of treatment until death. RESULTS: Patients initially treated with transdermal fentanyl started their strong opioid regime 8.5 years after diagnosis compared to 6.4 years after diagnosis in those who started SR morphine. This equates to an overall survival period from diagnosis of 8.8 years and 7.4 years respectively. Nevertheless, the total NHS cost of palliative care was similar between treatment groups, ranging from a mean 3087-3462 pounds per patient. Hospitalisation accounted for up to 71% of the total cost and opioids accounted for up to a further 17%. Less than one-third of patients received 4-hourly morphine as part of their initial opioid treatment despite UK guidelines recommending that moderate-to-severe pain should always be managed initially with an immediate-release preparation. Additionally, patients who received transdermal fentanyl as part of their initial treatment received significantly more laxative prescriptions than patients who started with SR morphine. CONCLUSIONS: SR morphine and transdermal fentanyl seem to be used in different situations. The results also confirm previous findings that pain management in cancer patients is often sub-optimal. The low contribution of opioids to the overall costs indicates that this should not be an obstacle to starting this aspect of palliative care earlier in disease progression. This characterisation of the resource implications of using SR morphine and transdermal fentanyl should enable purchasers and providers to optimise the availability of strong opioids for cancer patients on medical, economic and humanitarian grounds. PMID- 15801999 TI - Modelling the cost implications of using carboxymethylcellulose dressing compared with gauze in the management of surgical wounds healing by secondary intention in the US and UK. AB - OBJECTIVE: To estimate the costs of using carboxymethyl cellulose dressing (CMCD; Aquacel* Hydrofiber) compared to gauze in managing surgical wounds healing by secondary intention in the US and UK. STUDY DESIGN: This was a modelling study performed from the perspective of payers (i.e. the hospital and community sector in the US and the National Health Service (NHS) in the UK). METHODS: Clinical outcomes attributable to managing surgical wounds healing by secondary intention with gauze were obtained from the published literature in the English language. There were no published studies on wounds healing by secondary intention with CMCD. Hence, the analysis conservatively assumed that wound healing rates associated with gauze would be the same for CMCD. These data were combined with resource utilisation estimates derived from a panel of clinicians enabling us to perform decision modelling. The models were used to determine the expected direct healthcare costs eight weeks after the surgical wounds were dressed by CMCD or gauze and left to heal by secondary intention in the US and UK. RESULTS: All wounds are expected to heal within eight weeks, irrespective of dressing. Managing abscesses and other surgical wounds with CMCD instead of gauze in the US is expected to reduce costs by 4% in both wound types (i.e. $247 and $507 respectively) per patient over eight weeks. In the UK, managing abscesses and other surgical wounds with CMCD instead of gauze is expected to reduce costs by 30% (574 pounds) and 12% (581 pounds) respectively per patient over eight weeks. The lower cost of managing CMCD-treated patients is due to decreased nursing costs associated with a lower frequency of CMCD changes compared to gauze dressing changes. CONCLUSION: Dressing surgical wounds healing by secondary intention with CMCD instead of gauze is expected to lead to a reduction in healthcare costs in both the US and UK. Hence, the purchase price of a dressing is not indicative of the cost effectiveness of a given method of surgical wound care. PMID- 15802000 TI - Differences in hypoglycemia event rates and associated cost-consequence in patients initiated on long-acting and intermediate-acting insulin products. AB - OBJECTIVE: To compare hypoglycemia event rates in patients initiated on long acting insulin analog (glargine) or intermediate-acting insulin (NPH) and to analyze the associated cost-consequence from a managed care perspective. STUDY DESIGN: A retrospective analysis of pharmacy and medical claims and electronic laboratory result data using a southeastern United States managed care health plan. METHODS: Patients newly initiated on glargine or NPH between July 1, 2000 and August 31, 2002 were included. Hypoglycemia events were identified from medical claims by their ICD-9CM codes. Multivariable techniques were used to compare hypoglycemia event rates between cohorts. RESULTS: A total of 1434 patients were eligible (glargine = 310, NPH = 1124). The mean age was 53 years +/ 17 years and 51% of patients were male. The mean treatment duration was 8.6 months +/- 4.5 months. Multivariate analyses showed that patients in the NPH group had a higher hypoglycemia event rate than the glargine group (18.3 versus 7.3 per 100 patients per year; p = 0.009). The number needed to treat (glargine versus NPH) to avoid one hypoglycemia event per patient per year was nine patients at an A1C of 7%. The mean annual index medication cost was $47 more for glargine ($390) than for NPH ($343) per patient per year (p = 0.042). The mean cost per hypoglycemia event was $1087 (95% CI: $764-$1409). CONCLUSIONS: Patients treated with glargine had significantly lower hypoglycemia event rates compared to the NPH group. The risk difference indicated that one hypoglycemia event would be avoided for every nine patients treated with glargine instead of NPH. The cost increase associated with treating nine patients with glargine rather than NPH is less than the cost of treating one hypoglycemia event. In this population, the savings associated with reduced hypoglycemic events more than offset the increased acquisition cost associated with glargine. PMID- 15802001 TI - Have we forgotten the pivotal role of high-density lipoprotein cholesterol in atherosclerosis prevention? AB - The critical importance of high-density lipoprotein cholesterol (HDL-C) as an independent and predictive factor for coronary heart disease (CHD) has been increasingly recognised in treatment guidelines for prevention and treatment of cardiovascular disease. The association of low HDL-C with an increased incidence of CHD implies a critical role for raising HDL-C in protection against atherosclerotic disease. HDL-C appears to exert this effect via a number of mechanisms. HDL-C is involved in reverse cholesterol transport, prevents endothelial dysfunction and has anti-inflammatory, anti-oxidant and antithrombotic properties. Therapeutic interventions that increase HDL-C include statins, fibrates and nicotinic acid. Of these, nicotinic acid raises HDL-C more effectively than either statin or fibrate therapy and has been proven to reduce cardiovascular events in monotherapy studies. Preliminary clinical studies have shown that addition of nicotinic acid to primary statin therapy is safe, has proven beneficial effects on atherosclerosis and may also reduce the incidence of major coronary events. The available clinical evidence suggests that addition of nicotinic acid to primary lipid-lowering therapy has an important atheroprotective role in patients with or at risk of developing CHD. PMID- 15802002 TI - The involvement of professional medical writers in medical publications. AB - In this editorial, we comment on the European Medical Writers Association (EMWA) guidelines and the accompanying Delphi study published in this issue of Current Medical Research and Opinion. These articles deal with 'ghost writing' and the role of professional writers. We propose that the interaction between professional writers and authors is defined along the following principles: Guarantee: are the authors guarantors of the article? Advice: was the professional writer 'advised' by the author(s) before, as well as after, starting the assignment? Were the overall conclusions defined by the author(s)? Transparency: the contribution of professional writers should be acknowledged (we provide a draft statement). Expertise: does the professional writer have sufficient knowledge in the relevant specific field? The initials of these headings form the word ' GATE '. Therefore, we called them the ' GATE principles '. We also discuss suggestions for the accreditation of professional writers in specific fields. Professional writers play a useful role but this has to be clearly defined so as to achieve high ethical and scientific standards. PMID- 15802003 TI - European Medical Writers Association (EMWA) guidelines on the role of medical writers in developing peer-reviewed publications. AB - BACKGROUND: Many papers in biomedical journals are drafted not by the named authors, but by professional medical writers working under the direction of those authors, usually funded by pharmaceutical companies. Although this practice can improve both the quality and speed of publications, it has attracted controversy as a result of concerns about the inappropriate influence of pharmaceutical companies. OBJECTIVES: To define ethical standards for professional medical writers who prepare papers for publication in medical journals. CONSENSUS METHODS: Guidelines were drafted after a 4-round Delphi consultation among a group of experienced medical writers. The guidelines were then further refined by seeking comments on the draft from a range of interested parties. FINDINGS AND CONCLUSIONS: The guidelines stress the importance of respecting widely recognised authorship criteria, and in particular of ensuring that those listed as named authors have full control of the content of papers. The role of medical writers must be transparent, which normally means a mention in the acknowledgements section, together with a statement about funding. Writers and authors must have access to relevant data while writing papers. Medical writers have professional responsibilities to ensure that the papers they write are scientifically valid and are written in accordance with generally accepted ethical standards. PMID- 15802004 TI - Maximum likelihood topographic map formation. AB - We introduce a new unsupervised learning algorithm for kernel-based topographic map formation of heteroscedastic gaussian mixtures that allows for a unified account of distortion error (vector quantization), log-likelihood, and Kullback Leibler divergence. PMID- 15802005 TI - On convergence conditions of an extended projection neural network. AB - The output trajectory convergence of an extended projection neural network was developed under the positive definiteness condition of the Jacobian matrix of nonlinear mapping. This note offers several new convergence results. The state trajectory convergence and the output trajectory convergence of the extended projection neural network are obtained under the positive semidefiniteness condition of the Jacobian matrix. Comparison and illustrative examples demonstrate applied significance of these new results. PMID- 15802006 TI - Memorization and association on a realistic neural model. AB - A central open question of computational neuroscience is to identify the data structures and algorithms that are used in mammalian cortex to support successive acts of the basic cognitive tasks of memorization and association. This letter addresses the simultaneous challenges of realizing these two distinct tasks with the same data structure, and doing so while respecting the following four basic quantitative parameters of cortex: the neuron number, the synapse number, the synapse strengths, and the switching times. Previous work has not succeeded in reconciling these opposing constraints, the low values of synapse strengths that are typically observed experimentally having contributed a particular obstacle. In this article, we describe a computational scheme that supports both memory formation and association and is feasible on networks of model neurons that respect the widely observed values of the four quantitative parameters. Our scheme allows for both disjoint and shared representations. The algorithms are simple, and in one version both memorization and association require just one step of vicinal or neighborly influence. The issues of interference among the different circuits that are established, of robustness to noise, and of the stability of the hierarchical memorization process are addressed. A calculus therefore is implied for analyzing the capabilities of particular neural systems and subsystems, in terms of their basic numerical parameters. PMID- 15802007 TI - Effects of noisy drive on rhythms in networks of excitatory and inhibitory neurons. AB - Synchronous rhythmic spiking in neuronal networks can be brought about by the interaction between E-cells and Icells (excitatory and inhibitory cells). The I cells gate and synchronize the E-cells, and the E-cells drive and synchronize the I-cells. We refer to rhythms generated in this way as PING (pyramidal interneuronal gamma) rhythms. The PING mechanism requires that the drive I(I) to the I-cells be sufficiently low; the rhythm is lost when I(I) gets too large. This can happen in at least two ways. In the first mechanism, the I-cells spike in synchrony, but get ahead of the E-cells, spiking without being prompted by the E-cells. We call this phase walkthrough of the I-cells. In the second mechanism, the I-cells fail to synchronize, and their activity leads to complete suppression of the E-cells. Noisy spiking in the E-cells, generated by noisy external drive, adds excitatory drive to the I-cells and may lead to phase walkthrough. Noisy spiking in the I-cells adds inhibition to the E-cells and may lead to suppression of the E-cells. An analysis of the conditions under which noise leads to phase walkthrough of the I-cells or suppression of the E-cells shows that PING rhythms at frequencies far below the gamma range are robust to noise only if network parameter values are tuned very carefully. Together with an argument explaining why the PING mechanism does not work far above the gamma range in the presence of heterogeneity, this justifies the "G" in "PING." PMID- 15802008 TI - Supervised learning through neuronal response modulation. AB - Neural networks that are trained to perform specific tasks must be developed through a supervised learning procedure. This normally takes the form of direct supervision of synaptic plasticity. We explore the idea that supervision takes place instead through the modulation of neuronal excitability. Such supervision can be done using conventional synaptic feedback pathways rather than requiring the hypothetical actions of unknown modulatory agents. During task learning, supervised response modulation guides Hebbian synaptic plasticity indirectly by establishing appropriate patterns of correlated network activity. This results in robust learning of function approximation tasks evenwhenmultiple output units representing different functions share large amounts of common input. Reward based supervision is also studied, and a number of potential advantages of neuronal response modulation are identified. PMID- 15802009 TI - The combined effects of inhibitory and electrical synapses in synchrony. AB - Recent experimental results have shown that GABAergic interneurons in the central nervous system are frequently connected via electrical synapses. Hence, depending on the area or the subpopulation, interneurons interact via inhibitory synapses or electrical synapses alone or via both types of interactions. The theoretical work presented here addresses the significance of these different modes of interactions for the interneuron networks dynamics. We consider the simplest system in which this issue can be investigated in models or in experiments: a pair of neurons, interacting via electrical synapses, inhibitory synapses, or both, and activated by the injection of a noisy external current. Assuming that the couplings and the noise are weak, we derive an analytical expression relating the cross-correlation (CC) of the activity of the two neurons to the phase response function of the neurons. When electrical and inhibitory interactions are not too strong, they combine their effect in a linear manner. In this regime, the effect of electrical and inhibitory interactions when combined can be deduced knowing the effects of each of the interactions separately. As a consequence, depending on intrinsic neuronal properties, electrical and inhibitory synapses may cooperate, both promoting synchrony, or may compete, with one promoting synchrony while the other impedes it. In contrast, for sufficiently strong couplings, the two types of synapses combine in a nonlinear fashion. Remarkably, we find that in this regime, combining electrical synapses with inhibition amplifies synchrony, whereas electrical synapses alone would desynchronize the activity of the neurons. We apply our theory to predict how the shape of the CC of two neurons changes as a function of ionic channel conductances, focusing on the effect of persistent sodium conductance, of the firing rate of the neurons and the nature and the strength of their interactions. These predictions may be tested using dynamic clamp techniques. PMID- 15802010 TI - Spikernels: predicting arm movements by embedding population spike rate patterns in inner-product spaces. AB - Inner-product operators, often referred to as kernels in statistical learning, define a mapping from some input space into a feature space. The focus of this letter is the construction of biologically motivated kernels for cortical activities. The kernels we derive, termed Spikernels, map spike count sequences into an abstract vector space in which we can perform various prediction tasks. We discuss in detail the derivation of Spikernels and describe an efficient algorithm for computing their value on any two sequences of neural population spike counts. We demonstrate the merits of our modeling approach by comparing the Spikernel to various standard kernels in the task of predicting hand movement velocities from cortical recordings. All of the kernels that we tested in our experiments outperform the standard scalar product used in linear regression, with the Spikernel consistently achieving the best performance. PMID- 15802011 TI - Memory capacity of balanced networks. AB - We study the problem of memory capacity in balanced networks of spiking neurons. Associative memories are represented by either synfire chains (SFC) or Hebbian cell assemblies (HCA). Both can be embedded in these balanced networks by a proper choice of the architecture of the network. The size w(E) of a pool in an SFC or of an HCA is limited from below and from above by dynamical considerations. Proper scaling of w(E) by radicalK, where K is the total excitatory synaptic connectivity, allows us to obtain a uniform description of our system for any given K. Using combinatorial arguments, we derive an upper limit on memory capacity. The capacity allowed by the dynamics of the system, alpha(c), is measured by simulations. For HCA, we obtain alpha(c) of order 0.1, and for SFC, we find values of order 0.065. The capacity can be improved by introducing shadow patterns, inhibitory cell assemblies that are fed by the excitatory assemblies in both memory models. This leads to a doubly balanced network, where, in addition to the usual global balancing of excitation and inhibition, there exists specific balance between the effects of both types of assemblies on the background activity of the network. For each of the memory models and for each network architecture, we obtain an allowed region (phase space) for w(E)/ radicalK in which the model is viable. PMID- 15802012 TI - On the capabilities of higher-order neurons: a radial basis function approach. AB - Higher-order neurons with k monomials in n variables are shown to have Vapnik Chervonenkis (VC) dimension at least nk + 1. This result supersedes the previously known lower bound obtained via k-term monotone disjunctive normal form (DNF) formulas. Moreover, it implies that the VC dimension of higher-order neurons with k monomials is strictly larger than the VC dimension of k-term monotone DNF. The result is achieved by introducing an exponential approach that employs gaussian radial basis function neural networks for obtaining classifications of points in terms of higher-order neurons. PMID- 15802013 TI - Estimating the posterior probabilities using the k-nearest neighbor rule. AB - In many pattern classification problems, an estimate of the posterior probabilities (rather than only a classification) is required. This is usually the case when some confidence measure in the classification is needed. In this article, we propose a new posterior probability estimator. The proposed estimator considers the K-nearest neighbors. It attaches a weight to each neighbor that contributes in an additive fashion to the posterior probability estimate. The weights corresponding to the K-nearest-neighbors (which add to 1) are estimated from the data using a maximum likelihood approach. Simulation studies confirm the effectiveness of the proposed estimator. PMID- 15802015 TI - Genetic and epigenetic changes of components affecting the WNT pathway in colorectal carcinomas stratified by microsatellite instability. AB - An unselected series of 310 colorectal carcinomas, stratified according to microsatellite instability (MSI) and DNA ploidy, was examined for mutations and/or promoter hypermethylation of five components of the WNT signaling cascade [APC, CTNNB1 (encoding beta-catenin), AXIN2, TCF4, and WISP3] and three genes indirectly affecting this pathway [CDH1 (encoding E-cadherin), PTEN, and TP53]. APC and TP53 mutations were each present more often in microsatellite-stable (MSS) tumors than in those with MSI (P < .001 for both). We confirmed that the aneuploid MSS tumors frequently contained TP53 mutations (P < .001), whereas tumors with APC mutations and/or promoter hypermethylation revealed no associations to ploidy. Mutations in APC upstream of codons 1020 to 1169, encoding the beta-catenin binding site, were found in 15/144 mutated tumors and these patients seemed to have poor clinical outcome (P = .096). Frameshift mutations in AXIN2, PTEN, TCF4, and WISP3 were found in 20%, 17%, 46%, and 28% of the MSI tumors, respectively. More than half of the tumors with heterozygote mutations in AXIN2 were concurrently mutated in APC. The present study showed that more than 90% of all samples had alteration in one or more of the genes investigated, adding further evidence to the vital importance of activated WNT signaling in colorectal carcinogenesis. PMID- 15802016 TI - Ferritin as an endogenous MRI reporter for noninvasive imaging of gene expression in C6 glioma tumors. AB - The heavy chain of murine ferritin, an iron storage molecule with ferroxidase activity, was developed as a novel endogenous reporter for the detection of gene expression by magnetic resonance imaging (MRI). Expression of both enhanced green fluorescent protein (EGFP) and influenza hemagglutinin (HA)-tagged ferritin were tightly coregulated by tetracycline (TET), using a bidirectional expression vector. C6 cells stably expressing a TET-EGFP-HA-ferritin construct enabled the dynamic detection of TET-regulated gene expression by MRI, followed by independent validation using fluorescence microscopy and histology. MR relaxation rates were significantly elevated both in vitro and in vivo on TET withdrawal, and were consistent with induced expression of ferritin and increase in intracellular iron content. Hence, overexpression of ferritin was sufficient to trigger cellular response, augmenting iron uptake to a degree detectable by MRI. Application of this novel MR reporter gene that generates significant contrast in the absence of exogenously administered substrates opens new possibilities for noninvasive molecular imaging of gene expression by MRI. PMID- 15802017 TI - Liposome-encapsulated prednisolone phosphate inhibits growth of established tumors in mice. AB - Glucocorticoids can inhibit solid tumor growth possibly due to an inhibitory effect on angiogenesis. The antitumor effects of the free drugs have only been observed using treatment schedules based on high and frequent dosing for prolonged periods of time. As long-circulating liposomes accumulate at sites of malignancy, we investigated the tumor-inhibiting potential of liposome encapsulated prednisolone phosphate. Liposomal prednisolone phosphate could inhibit tumor growth dose-dependently, with 80% to 90% tumor growth inhibition of subcutaneous B16.F10 melanoma and C26 colon carcinoma murine tumor models at 20 mg/kg by single or weekly doses. Prednisolone phosphate in the free form was completely ineffective at this low-frequency treatment schedule, even when administered at a dose of 50 mg/kg. In vitro studies did not show an inhibitory effect of prednisolone (phosphate) on tumor cell, nor on endothelial cell proliferation. Histologic evaluation revealed that liposomal prednisolone phosphate-treated tumors contained a center with areas of picnotic/necrotic cells, which were not apparent in untreated tumors or tumors treated with the free drug. In conclusion, the present study shows potent antitumor effects of liposomal formulations of glucocorticoids in a low dose and low-frequency schedule, offering promise for liposomal glucocorticoids as novel antitumor agents. PMID- 15802019 TI - pRb-Independent growth arrest and transcriptional regulation of E2F target genes. AB - The retinoblastoma tumor suppressor (pRb) has traditionally been studied as a negative regulator of cell cycle progression through its interactions with the E2F family of transcription factors. Utilizing prostate epithelial cell lines established from Rb+/+ and Rb-/- prostate tissues, we previously demonstrated that Rb-/- epithelial cells were not transformed and retained the ability to differentiate in vivo despite the lack of pRb. To further study the effects of pRb loss in an epithelial cell population, we utilized oligonucleotide microarrays to identify any pRb-dependent transcriptional regulation during serum depletion-induced growth arrest. These studies identified 120 unique transcripts regulated by growth arrest in Rb+/+ cells. In these wild-type cells, the majority (80%) of altered transcripts were downregulated, including 40 previously identified E2F target genes. Although the transcriptional repression of E2F target genes is characteristic of pRb pocket protein family activity, further analysis revealed that, compared to Rb+/+ cells, Rb-/- cells exhibited a nearly identical response for all transcripts including those of E2F target genes. These findings demonstrate that pRb is not strictly required for the vast majority of transcriptional alterations associated with growth arrest. PMID- 15802018 TI - Resveratrol and estradiol exert disparate effects on cell migration, cell surface actin structures, and focal adhesion assembly in MDA-MB-231 human breast cancer cells. AB - Resveratrol, a grape polyphenol, is thought to be a cancer preventive, yet its effects on metastatic breast cancer are relatively unknown. Since cancer cell invasion is dependent on cell migration, the chemotactic response of MDA-MB-231 metastatic human breast cancer cells to resveratrol, estradiol (E2), or epidermal growth factor (EGF) was investigated. Resveratrol decreased while E2 and EGF increased directed cell migration. Resveratrol may inhibit cell migration by altering the cytoskeleton. Resveratrol induced a rapid global array of filopodia and decreased focal adhesions and focal adhesion kinase (FAK) activity. E2 or EGF treatment did not affect filopodia extension but increased lamellipodia and associated focal adhesions that are integral for cell migration. Combined resveratrol and E2 treatment resulted in a filopodia and focal adhesion response similar to resveratrol alone. Combined resveratrol and EGF resulted in a lamellipodia and focal adhesion response similar to EGF alone. E2 and to a lesser extent resveratrol increased EGFR activity. The cytoskeletal changes and EGFR activity in response to E2 were blocked by EGFR1 inhibitor indicating that E2 may increase cell migration via crosstalk with EGFR signaling. These data suggest a promotional role for E2 in breast cancer cell migration but an antiestrogenic, preventative role for resveratrol. PMID- 15802020 TI - Cellular basis of antiproliferative and antitumor activity of the novel camptothecin derivative, gimatecan, in bladder carcinoma models. AB - To investigate the cellular/molecular basis of the activity of a novel lipophilic camptothecin, gimatecan (ST1481), against slowly proliferating cells, we performed a comparative study of topotecan and gimatecan in human bladder cancer models (HT1376 and MCR). Gimatecan was significantly more effective than topotecan in inhibiting the growth of HT1376 tumor, thus reflecting antiproliferative potency. In both HT1376 and MCR cells, gimatecan caused a persistent S-phase arrest, indicating an efficient DNA damage checkpoint. This response was consistent with a cytostatic effect, because no evidence of apoptosis was detected. In contrast to gimatecan, topotecan at equitoxic concentrations caused an early and persistent downregulation of topoisomerase I. Modulation of protein level could not be solely ascribed to the proteasome mediated degradation of the enzyme because the proteasome inhibitor PS341 sensitized MCR but not HT1376 cells to camptothecins, suggesting alternative mechanisms of drug-induced topoisomerase I downregulation. Indeed, the two camptothecins caused a differential inhibition of topoisomerase I transcription, which is more marked in topotecan-treated cells. The HT1376 model was more sensitive to this immediate decrease of mRNA level. Our data document a marked antitumor activity of gimatecan against a bladder carcinoma model. A limited downregulation of topoisomerase I by gimatecan provides additional insights into the cellular basis of drug potency. PMID- 15802021 TI - Sensitization for anticancer drug-induced apoptosis by betulinic Acid. AB - We previously described that betulinic acid (BetA), a naturally occurring pentacyclic triterpenoid, induces apoptosis in tumor cells through the mitochondrial pathway. Here, for the first time, we provide evidence that BetA cooperated with anticancer drugs to induce apoptosis and to inhibit clonogenic survival of tumor cells. Combined treatment with BetA and anticancer drugs acted in concert to induce loss of mitochondrial membrane potential and the release of cytochrome c and Smac from mitochondria, resulting in activation of caspases and apoptosis. Overexpression of Bcl-2, which blocked mitochondrial perturbations, also inhibited the cooperative effect of BetA and anticancer drugs, indicating that cooperative interaction involved the mitochondrial pathway. Notably, cooperation of BetA and anticancer drugs was found for various cytotoxic compounds with different modes of action (e.g., doxorubicin, cisplatin, Taxol, VP16, or actino-mycin D). Importantly, BetA and anticancer drugs cooperated to induce apoptosis in different tumor cell lines, including p53 mutant cells, and also in primary tumor cells, but not in human fibroblasts indicating some tumor specificity. These findings indicate that using BetA as sensitizer in chemotherapy-based combination regimens may be a novel strategy to enhance the efficacy of anticancer therapy, which warrants further investigation. PMID- 15802022 TI - Therapeutic efficacy of adenoviral-mediated p53 gene transfer is synergistically enhanced by combined use of antisense oligodeoxynucleotide targeting clusterin gene in a human bladder cancer model. AB - To establish a more effective therapeutic strategy against advanced bladder cancer, we investigated the effects of combined treatment with antisense (AS) oligodeoxynucleotide (ODN) targeting the anti-apoptotic gene clusterin and adenoviral-mediated p53 gene transfer (Ad5CMV-p53) using the human bladder cancer KoTCC-1 model. Clusterin expression in KoTCC-1 cells was highly upregulated by Ad5CMV-p53 treatment; however, AS clusterin ODN treatment further suppressed clusterin expression in KoTCC-1 cells after Ad5CMV-p53 treatment. AS clusterin ODN treatment synergistically enhanced the cytotoxic effect of Ad5CMV-p53, and DNA fragmentation characteristic of apoptosis was observed only after combined treatment with AS clusterin ODN and Ad5CMV-p53, but not after treatment with either agent alone. Administration of AS clusterin ODN and Ad5CMV-p53 into nude mice resulted in a significant inhibition of KoTCC-1 tumor growth as well as lymph node metastases compared to administration of either agent alone. Furthermore, combined treatment with AS clusterin ODN, Ad5CMV-p53, and cisplatin completely eradicated KoTCC-1 tumors and lymph node metastases in 60% and 100% of mice, respectively. These findings suggest that combined treatment with AS clusterin ODN and Ad5CMV-p53 could be a novel strategy to inhibit bladder cancer progression, and that further additional use of a chemotherapeutic agent may substantially enhance the efficacy of this combined regimen. PMID- 15802024 TI - Reflective testing: the practice of adding on tests by laboratory staff. PMID- 15802023 TI - Semaphorin SEMA3F has a repulsing activity on breast cancer cells and inhibits E cadherin-mediated cell adhesion. AB - Previously, we demonstrated that loss of SEMA3F, a secreted semaphorin encoded in 3p21.3, is associated with higher stages in lung cancer and primary tumor cells studied with anti-vascular endothelial growth factor (VEGF) and SEMA3F antibodies. In vitro, SEMA3F inhibits cell spreading; this activity is opposed by VEGF. These results suggest that VEGF and SEMA3F compete for binding to their common neuropilin receptor. In the present report, we investigated the attractive/repulsive effects of SEMA3F on cell migration when cells were grown in a three-dimensional system and exposed to a SEMA3F gradient. In addition, we adapted the neurobiologic stripe assay to analyze the migration of tumor cells in response to SEMA3F. In the motile breast cancer cell line C100, which expresses both neuropilin-1 (NRP1) and neuropilin-2 (NRP2) receptors, SEMA3F had a repulsive effect, which was blocked by anti-NRP2 antibody. In less motile MCF7 cells, which express only NRP1, SEMA3F inhibited cell contacts with loss of membrane-associated E-cadherin and beta-catenin without motility induction. Cell spreading and proliferation were reduced. These results support the concept that in a first step during tumorigenesis, normal tissues expressing SEMA3F would try to prevent tumor cells from spreading and attaching to the stroma for further implantation. PMID- 15802025 TI - The new General Medical Services contract: opportunity or threat? PMID- 15802026 TI - Impact of the new General Medical Services contract on the clinical laboratory. AB - The new General Medical Services contract that has been adopted in England and Wales, with similar contracts in Scotland and Northern Ireland, proposes far reaching changes to the way in which general practitioners practise and are remunerated. This includes a range of quality indicators, which pay particular attention to chronic disease management. Among other things, the quality indicators require certain laboratory tests to be performed or result targets to be met in defined proportions of patients in particular disease categories in order to qualify for quality payments. This is a novel experiment, which may focus general practitioner activity on specific areas of patient management. It has significant potential impact on laboratories: testing activity in some areas may be boosted considerably; assay performance, notably bias, will have direct potential impact on practice earnings and any shift in clinical priorities may have future effects on assay development and repertoires as these become partly financially driven by practice income. The evidence base and research of the available guidance behind the indicators is sketchy in places and although there are 76 indicators several areas of practice are not addressed. The probable further increase in laboratory activity it may generate highlights the need for laboratories to continue to assist in promoting best practice in test requesting. PMID- 15802027 TI - Susceptibility of glomerular filtration rate estimations to variations in creatinine methodology: a study in older patients. AB - BACKGROUND: It is recommended that measurement of serum creatinine should be supplemented with a creatinine-based estimation of glomerular filtration rate (GFR). The influence of creatinine methodology on these estimates is not always appreciated. We have studied differences in creatinine methods and their influence on GFR estimation specifically in older people. METHODS: In all, 46 older patients (mean age 80 y, range 69-92 y) with predominantly mild or moderate kidney disease were studied. Serum creatinine was measured using a rate Jaffe method and two different enzymatic methods. Isotope dilution mass spectrometry served as the reference creatinine method. GFR was estimated using both the Modification of Diet in Renal Disease (MDRD) and Cockcroft and Gault formulae: a 51Cr-EDTA GFR estimation served as the reference GFR method. RESULTS: Both enzymatic methods produced creatinine results that were significantly different (P<0.001) from the reference method. The Jaffe method over- and underestimated creatinine at low and high concentrations, respectively. The most likely explanation for these differences relates to standardization of the assays. Irrespective of creatinine method, the Cockroft and Gault formula tended to underestimate GFR, and the MDRD formula to overestimate GFR. Use of the differing creatinine methods to estimate GFR produced predictable biases of the estimate, with mean GFR estimates varying by 14% across the creatinine methods. CONCLUSION: Estimates of GFR depend critically upon the accuracy and precision of the creatinine measurement used in their calculation. PMID- 15802028 TI - The comparative analytical performance of four troponin I assays at low concentration. AB - BACKGROUND: Low troponin concentrations have been shown to be informative in the prognosis of acute coronary syndrome. We have investigated the analytical performance of four commonly used cardiac troponin I methods at concentrations approaching their analytical limit of detection. METHOD: We assayed 167 patient samples within 24 h of collection using the Beckman Coulter AccuTnI, Dade Behring Dimension, Abbott AxSYM and Bayer Centaur methods and compared their relative analytical performance. RESULTS: Of the four assays compared, the AccuTnI was observed to have greater sensitivity at low concentrations. Using the limit of detection as the threshold, the Beckman assay showed superior performance at concentrations corresponding to a 20% coefficient of variation (CV), the Dade assay had a similar performance; and at concentrations corresponding to 10% CV most assays provide similar information. CONCLUSION: The newer or recently modified assays such as the Beckman Coulter AccuTnI and Dade Behring assays are best able to identify very low concentrations of troponin. PMID- 15802029 TI - Interpretation of alpha-fetoprotein concentrations in cerebrospinal fluid of infants. AB - BACKGROUND: There are no published reference intervals for concentrations of alpha-fetoprotein (AFP) in the cerebrospinal fluid (CSF) of normal infants. The presence of abnormal concentrations of AFP in plasma or CSF may indicate the presence of a teratoma or a germ cell tumour with yolk sac elements. We measured CSF AFP in infants who did not have malignancy in order to determine its reference intervals. METHODS: AFP was measured in the CSF and/or plasma in 128 infants. Of these, 91 infants had CSF AFP measurements, 94 infants had plasma AFP measurements and in 60 infants AFP concentrations were determined in paired CSF and plasma samples. The patients ranged in age from 1 to 110 days. Both CSF and plasma AFP concentrations were measured by a microparticle enzyme immunoassay using an AxSYM analyser. RESULTS: Using ages corrected for prematurity, the median CSF AFP concentration for babies -69 to 31 days old was 61 kIU/L (5th-95th centile: 2-889 kIU/L), while the median CSF AFP concentration for infants 32 to 110 days was 1.2 kIU/L (5th-95th centile: 0.1-12.5 kIU/L). By age 6 weeks, the concentrations were close to those found in adult plasma and all CSF AFP concentrations from infants with a corrected age over 2 months were <3 kIU/L. CONCLUSION: We have defined reference intervals for CSF AFP concentrations in infants. These results may assist in the diagnosis of CNS tumours, particularly congenital CNS tumours containing yolk sac elements. PMID- 15802030 TI - First trimester maternal serum screening for Down's syndrome: an evaluation of the DPC Immulite 2000 free beta-hCG and pregnancy-associated plasma protein-A assays. AB - BACKGROUND: Recent NICE Guidelines have emphasized the need to have in place by 2007 the capability of offering screening to all women in the first trimester using a combination of maternal age with the ultrasound marker nuchal translucency thickness (NT) and the maternal serum biochemical markers free beta hCG and pregnancy-associated plasma protein-A (PAPP-A). Laboratories will therefore need to consider how to introduce the biochemical component of screening. With the recent launch of these assays on the DPC Immulite 2000 platform, it is appropriate and timely to investigate their clinical and analytical performance on a high throughput immunoassay analyser. METHODS: Within run and between-day precision was assessed in the normal way. Bias was assessed by comparing samples from normal pregnancies (n=813) and pregnancies with Down's syndrome (n=60) run on both the DPC system and our routine Kryptor system. Gestational day-specific medians for each marker were calculated from the unaffected population. Mathematical modelling was used to predict the clinical performance of the two markers. RESULTS: The within-run coefficient of variation (CV) was around 3.5% and between-day CV was around 6-8% for both assays. Comparison with EQA samples showed a 2% positive bias against the ALTM for free beta-hCG and a 21% positive bias for PAPP-A, which reduced to 11% when compared with the Kryptor method mean. When compared with the Kryptor, the DPC free beta hCG showed a significant concentration-related negative bias above concentrations of 50 IU/L in both unaffected and affected pregnancy samples. After conversion to MoM in the Down's syndrome group, the DPC free beta-hCG assay showed a MoM related bias at higher MoM values. For PAPP-A a positive MoM-related bias was also evident. In cases with Down's syndrome, the median free beta-hCG MoM was 1.703 for DPC and 1.698 by the Kryptor, which was not significantly different. For PAPP-A, the median MoM was 0.62 for DPC and 0.47 by the Kryptor, which was significantly (P=0.025) different. This difference, if real, is likely to result in a lower detection of cases with Down's syndrome. Statistical modelling techniques suggest that this would be 58% compared with the expected 67% when combined with maternal age and free beta-hCG. When combined with NT, however, this would be reduced to 85% compared with the 90% expected. CONCLUSIONS: While laboratories may find some organizational benefit from running first trimester Down's syndrome screening on such routine high-throughput immunoassay analysers, better clinical performance is likely to be achieved with alternative platforms to the DPC Immulite 2000. PMID- 15802031 TI - Japanese standard reference material for JDS Lot 2 haemoglobin A1c. I: Comparison of Japan Diabetes Society-assigned values to those obtained by the Japanese and USA domestic standardization programmes and by the International Federation of Clinical Chemistry reference laboratories. AB - BACKGROUND: The Committee on Standardization of Laboratory Testing Related to Diabetes Mellitus of the Japan Diabetes Society (JDS) previously recommended use of the primary calibrator (JDS Lot 1) prepared by the former Committee for Standardization of Glycohemoglobin for standardizing the measurement of haemoglobin A1c (HbA1c). Owing to the depletion of vials of Lot 1 in March 2001, the present committee certified a new reference material, Lot 2, now distributed by the Health Care Technology Foundation (HECTEF). The standardization programme for HbA1c measurement in Japan is currently based on Lot 2, which has values assigned from within Lot 1; the Lot 1 values were consensus values based on assays by laboratories in the Japanese national quality control programme. In this study, for the purpose of international comparison and standardization, Lot 2 was assayed by the JDS reference laboratories, the National Glycoprotein Standardization Program (NGSP) in the USA, and by reference laboratories approved by the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). METHOD: The HbA1c values of JDS Lot 2 were transferred from those assigned to Lot 1 using KO500, a high-resolution HPLC method, at three laboratories approved by the JDS committee. Subsequently, vials of JDS Lot 2 were shipped to and assayed by the NGSP in the USA and 10 IFCC reference laboratories. RESULT: The JDS-assigned HbA1c values (from Lot 1) are 4.04 for Level 1, 5.38 for Level 2, 7.32 for Level 3, 9.88 for Level 4, and 12.63 for Level 5, all expressed as a percentage of total haemoglobin. The values obtained by NGSP and the IFCC laboratories gave the following formulas: NGSP value(%)=JDS value(%)+0.3%; IFCC value(%)=1.068xJDS value(%)-1.741%. CONCLUSION: Although the values obtained by the IFCC laboratories are significantly lower than the values assigned to Lot 2 by the JDS, the relationship is linear. In addition, standardization of HbA1c based on JDS Lot 2 is currently at a satisfactory level in Japan. As a result, the reassignment of values for Lot 2 to agree with the IFCC values should be relatively easy and will be done after all relevant parties agree to the change. PMID- 15802032 TI - Japanese standard reference material JDS Lot 2 for haemoglobin A1c. II: Present state of standardization of haemoglobin A1c in Japan using the new reference material in routine clinical assays. AB - BACKGROUND: In 2001, the Committee on Standardization of Laboratory Testing Related to Diabetes Mellitus of the Japan Diabetes Society (JDS) prepared and certified a new reference material for haemoglobin A1c (HbA1c), Lot 2. The standardization programme for HbA1c measurement in Japan is currently based on Lot 2, although some laboratories still use the previous material (Lot 1). The values assigned to Lot 2 were based on the consensus values for Lot 1 and should give the same results. Therefore, there should be no difference in the measured values no matter which calibrators are used. The Committee conducted a domestic survey in order to confirm this relationship. METHOD: In November 2002, four samples for HbA1c assay were sent to 795 laboratories as part of a national survey in Japan. Assays were performed using the laboratories' routine clinical methods. The coefficients of variation (CVs) of the reported values from all laboratories for the samples were calculated in order to determine the current level of standardization in Japan. RESULTS: The overall CVs in the measured values for the four samples ranged from 2.7% to 4.0%. Values from laboratories using calibrators based on Lots 1 and 2 were similar. CONCLUSION: The present state of standardization for the routine measurement of HbA1c in Japan, as indicated by the 2002 survey, is excellent. This should aid in the eventual conversion of Lot 2 to IFCC-based values from the results of the 2002 national HbA1c survey. PMID- 15802033 TI - Plasma aluminium: a redundant test for patients on dialysis? AB - BACKGROUND: Aluminium toxicity as a cause of dementia, osteodystrophy and anaemia in patients receiving renal dialysis was first described in the 1970s and led to the regular monitoring of aluminium in plasma and dialysate water. However, aluminium phosphate binders have now been replaced by calcium-based binders or sevelamer and reverse osmosis (RO) water is used in the preparation of dialysate fluid. This has reduced the exposure of dialysis patients to aluminium and it is therefore opportune to review aluminium monitoring in patients undergoing regular renal dialysis. METHODS: Plasma and water aluminium results were audited over the period January 2000-January 2004, with results obtained from nine renal dialysis units in the UK. Patients with a plasma aluminium concentration in the toxic range (>3.7 micromol/L) were followed up by contacting the relevant consultant. RESULTS: Plasma aluminium results were collected on 1626 patients over the four year period (mean=0.47 micromol/L, median=0.3 micromol/L, range 0.07-30.26 micromol/L, n=5918). Forty-six patients had an aluminium concentration >3.7 micromol/L and nine were not retested. Only three patients had a repeat aluminium concentration >2.2 micromol/L, one being a result of desferrioxamine treatment, with no further clinical information available on the other two. All renal units are using RO water to prepare dialysate and aluminium-based phosphate binders are no longer prescribed. Only one of 212 RO water aluminium concentrations measured was >10 microg/L. Patients with clinical symptoms of overt aluminium toxicity were not identified in this population. CONCLUSION: The role of aluminium monitoring in long-term renal dialysis patients needs re-evaluation. Regular monitoring of plasma aluminium may not be required, but should be considered in any patient showing signs or symptoms of aluminium toxicity or exposed to a contaminated water supply. It is more important that RO water supplies are maintained and monitored. Environmental aluminium as a source of sample contamination should be considered and eliminated during blood collection and sample processing. PMID- 15802034 TI - Lymphocyte PC-1 activity in patients on maintenance haemodialysis treated with human erythropoietin and 1-alpha-D3. AB - BACKGROUND: Plasma cell differentiation antigen 1 (PC-1) is an inhibitor of insulinreceptor tyrosine-kinase. PC-1 content is elevated in muscle and adipose tissue from insulin-resistant subjects and its elevation correlates with in vivo insulin resistance. It is known that insulin resistance in uraemia may be improved with erythropoietin (EPO) and vitamin D therapy. Therefore, in this study the effects of human recombinant EPO and 1-alpha-D3 treatments on lymphocyte PC-1 expression in patients with end-stage renal failure on haemodialysis (HD) were investigated. METHODS: Lymphocyte basal, concanavalin A (Con A), and phorbol-12-myristate13-acetate (PMA)-stimulated PC-1 activity were investigated in HD patients before and after a two-month treatment with subcutaneous EPO (15 patients, 2000-3000 U thrice weekly) or oral 1-alpha-D3 (14 patients, 2 mug thrice weekly). Twenty-nine patients (16 men and 13 women), aged 22-68 years (49+/-7 years), on HD from 13 to 112 months, and 30 healthy controls participated in the study. None was obese and all had normal fasting plasma glucose. RESULTS: A two-month EPO treatment produced a 41% haematocrit increase, with a rise in haemoglobin from 6.51+/-0.18 g/dL to 9.69+/-0.14 g/dL. Basal lymphocyte PC-1 activity in HD patients was found to be significantly increased (P<0.005) over the level in healthy controls. Treatment of patients with EPO decreased unstimulated lymphocyte PC-1 activity to values significantly lower than before the treatment (P<0.001). Lymphocyte Con A and PMA-stimulated PC-1 activity in patients on HD was found to be slightly increased over the level in healthy controls, but significantly reduced (P<0.005 and 0.05, respectively) after the EPO treatment. A two-month pulse oral 1-alpha-D3 treatment increased haematocrit by 21% and raised haemoglobin from 7.11+/-0.32 g/dL to 8.80+/-0.39 g/dL. This treatment normalized serum alkaline phosphatase activity and slightly reduced serum parathyroid hormone concentration. PC-1 in unstimulated and PMA stimulated lymphocytes was unchanged, but significantly decreased (P<0.05) in Con A-stimulated lymphocytes after 1-alpha-D3 treatment. Fasting plasma glucose was not changed by the treatment. CONCLUSION: An increased lymphocyte PC-1 activity over control was found in HD patients. A two-month EPO therapy significantly decreased PC-1 activity to the control values, suggesting that an effect on PC-1 expression could be implicated in the amelioration of insulin resistance in uraemic patients treated with EPO. Treatment with pulse oral 1-alpha-D3 had an effect only on PC-1 of Con A-transformed lymphocytes of haemodialysed patients and requires further investigation. PMID- 15802035 TI - Troponin reporting in Australia in 2003. AB - BACKGROUND AND METHOD: This study uses a cross-sectional survey by questionnaire of Australia's pathology laboratories to describe how they report chemical markers for myocardial necrosis. RESULTS: The questionnaire was sent to 364 laboratories; 346 (95%) responded. Reporting data were obtained for 222 instruments used to analyse markers of myocardial necrosis, some 81% of all those in Australia. All laboratories use a troponin measurement. Reports from 132 analysers (59% of total) provide one cut-off point for the diagnosis of myocardial necrosis and 90 (41%) two cut-off points. The chosen cut-off points vary. The upper limit of 'healthy' is cited, alone, in 40 reports (18%); at this level, in 2002 no analysers predictably met required precision standards. Only 50% of all reports carry some explanatory comment; only 16% of all reports cite the source of the data. CONCLUSIONS: Troponin is used throughout Australia to help diagnose myocardial necrosis, but there is little order in the way the results are reported or interpreted. PMID- 15802036 TI - Carbonyl stress in chronic renal failure: the effect of haemodialysis. AB - Oxidative stress has been defined as a loss of balance between free radical production and the antioxidant systems. There have been many reports of increased production of oxidants and decreased levels of antioxidants in chronic renal failure (CRF) patients. An increase in oxidative stress may contribute to the development of oxidative protein damage in CRF. Our aim was to reveal oxidative modifications of plasma proteins by measuring 2,4-dinitrophenylhydrazine reactive carbonyl derivates (PCO), protein thiol (P-SH) and reduced glutathione (GSH) in predialytic uraemic and haemodialysed (HD) patients before and after dialysis. We included 20 predialytic uraemic patients, 20 HD patients and 20 healthy volunteers in our study. PCO concentration in predialytic uraemic patients increased compared with the concentration of the control group and this increase was more profound in HD patients. P-SH concentrations were significantly decreased in haemodialytic patients compared with those of controls. GSH level was higher in HD patients (both before and after dialysis). Increased PCO and decreased P-SH concentrations in all patient groups in comparison to the control subjects indicate increased protein oxidation. Our data in ESRD patients propose plasma protein carbonyl derivates and thiol concentrations as novel specific markers for oxidative protein damage. PMID- 15802037 TI - Conjugated bilirubin measurements in Gilbert's syndrome: method dependence. AB - BACKGROUND: Introducing new methodology often requires alteration to reference ranges and may cause inconvenience. Reagent suppliers may not have validated reference ranges quoted in their method sheets. When ratios of analytes are important, as for conjugated and unconjugated bilirubin, the combined changes can cause confusion. METHODS: The effect on bilirubin result interpretation following a change from Vitros (E950) dry slide technology to the Bayer ADVIA 1650 wet chemistry system was studied. RESULTS AND CONCLUSION: Over-estimation of conjugated bilirubin without an appropriate reference range can cause interpretative confusion. It is important to identify key patient groups likely to be affected by method changes well in advance. These need to be worked up in addition to reference range checks. It is unwise to rely on manufacturers for advice in this area. This report gives conjugated or direct bilirubin and total bilirubin values obtained using the above methods in nine patients with Gilbert's syndrome. PMID- 15802038 TI - The immunodetection of lipoprotein-bound amyloid-beta is attenuated because of the presence of lipids. AB - BACKGROUND: Amyloid-beta (Abeta) is found in circulation and cerebrospinal fluid (CSF), predominantly associated with lipoproteins. However, in a lipid environment it is possible that a masking of Abeta epitopes and/or an altered conformation of Abeta leads to an underestimation of Abeta concentrations. METHODS: We generated lipoprotein-like lipid emulsions containing a known amount of Abeta and compared immunoreactivity with an equimolar amount of Abeta solubilized in an aqueous medium. RESULTS: We found that Abeta exists primarily as a dimer and a monomer within an aqueous and lipid environment, respectively. We also showed that lipids bind tightly to Abeta, blocking detection of the monomeric form and substantially attenuating detection of the dimeric form of the protein. CONCLUSION: It is possible that studies that have quantified the concentration of Abeta in plasma and CSF may have significantly underestimated the pool of lipoprotein-bound Abeta. PMID- 15802039 TI - The importance of the interpretation of urine catecholamines is essential for the diagnosis and management of patient with dopamine-secreting paraganglioma. AB - Phaeochromocytoma or paraganglioma that exclusively secretes dopamine is very rare. This case illustrates its atypical presentation and the importance of interpretative reporting for urine catecholamines leading to the diagnosis and subsequent management of a patient with this condition. We report a 71-year-old Chinese woman with a large dopamine-secreting paraganglioma. She presented with low back pain for six months. On examination, a right abdominal mass was palpable incidentally. Her blood pressure was normal throughout. Serial 24-h urine collections for catecholamines showed enormous elevation of urine dopamine excretion to 80.7 micromol/day (normotensive:<2.6 m mol/day). However, the daily excretions of urine adrenaline and noradrenaline, as well as their metabolites were within their respective reference intervals. Good communication between chemical pathologists and physicians prompted the arrangement of the whole body 131I-meta-iodobenzylguanidine (MIBG) scintigraphy, which showed a large signal in the right upper quadrant of the abdomen corresponding to a large extra-adrenal tumour detected by both ultrasonography and computerized tomography (CT) of the abdomen. Histological section of the tumour tissue revealed paraganglioma, which stained positive for chromogranin and neuron-specific enolase. After four months, the patient presented with chest symptoms and CT of the thorax revealed multiple nodules. Lung metastases were suspected. However, follow-up urine catechola- mine and dopamine excretions were again within their respective normotensive reference intervals. A second MIBG scintigraphy was performed, but no specific uptake at either the thorax or the abdomen could be demonstrated. Fine-needle aspiration cytology using the thoracoscopic technique was performed and immunochemical staining of the biopsy specimen showed the presence of non- small-cell carcinoma of the lung. PMID- 15802040 TI - Recurrent dizzy spells: all in the head! AB - Hypoglycaemia is an uncommon manifestation of hypopituitarism in non-diabetic adults. A 47-year-old woman with Cushing's disease was treated with bilateral adrenalectomy and pituitary radiotherapy following unsuccessful trans-sphenoidal surgery. Twenty years later, she presented with a tonic-clonic seizure and recurrent symptoms suggestive of spontaneous hypoglycaemia. Her symptoms continued despite manipulation of her glucocorticoid replacement therapy. Recurrent severe hypoglycaemia resolved after identification and treatment of her concomitant growth hormone deficiency. In patients with hypopituitarism, undiagnosed growth hormone deficiency can influence glucose metabolism and predispose to severe hypoglycaemia. PMID- 15802041 TI - Phenotype variability in a daughter and father with mild osteogenesis imperfecta correlated with collagen and prolidase levels in cultured skin fibroblasts. AB - Studies of collagen biosynthesis and prolidase activity were performed on cultured skin fibroblasts obtained from a female patient and her father, who displayed variable phenotypes of mild osteogenesis imperfecta (OI). For comparison, the same studies were also performed on age-matched controls. Biosynthesis of collagen in fibroblasts of the less affected father was reduced to approximately 50% of control levels, whereas in cells of the more severely affected daughter, it was decreased to about 20% of control levels. Furthermore, the decrease in collagen synthesis in OI fibroblasts was accompanied by a parallel decrease in prolidase activity and expression of beta1 integrin and insulin-like growth factor-I (IGF-I) receptors recovered from the cells. Therefore, prolidase, as well as IGF-I and beta1 integrin receptors involved in collagen metabolism regulation, may represent important factors influencing OI phenotype. PMID- 15802042 TI - Copper deficiency. PMID- 15802045 TI - Piscator. PMID- 15802046 TI - Noninvasive assessment of gastric emptying by near-infrared fluorescence reflectance imaging in mice: pharmacological validation with tegaserod, cisapride, and clonidine. AB - Noninvasive near-infrared fluorescence reflectance imaging (FRI) is an in vivo technique to assess physiological and molecular processes in the intact organism. Here we describe a method to assess gastric emptying in mice. TentaGel beads with covalently bound cyanine dye (Cy5.5) conjugates as fluorescent probe were administered by oral gavage. The amount of intragastric beads/label was derived from the fluorescence signal intensity measured in a region of interest corresponding to the mouse stomach. The FRI signal intensity decreased as a function of time reflecting gastric emptying. In control mice, the gastric half emptying time was in agreement with literature data. Pharmacological modulation of gastric motility allowed the evaluation of the sensitivity of the FRI-based method. Gastric emptying was either stimulated or inhibited by treatment with the 5-HT(4) receptor agonists tegaserod (Zelnorm) and cisapride or the alpha(2) receptor agonist clonidine, respectively. Tegaserod and cisapride dose dependently accelerated gastric emptying. In contrast, clonidine dose-dependently delayed gastric emptying. In conclusion, FRI using fluorescently labeled beads allows the reliable determination of gastric emptying as well as the assessment of pharmacological interventions. The technique thus offers the potential to characterize molecular targets and pathways involved in physiological regulation and pharmacological modulation of gastric emptying. PMID- 15802047 TI - Quantitative analysis of micro-CT imaging and histopathological signatures of experimental arthritis in rats. AB - Micro-computed tomographic (micro-CT) imaging provides a unique opportunity to capture 3-D architectural information in bone samples. In this study of pathological joint changes in a rat model of adjuvant-induced arthritis (AA), quantitative analysis of bone volume and roughness were performed by micro-CT imaging and compared with histopathology methods and paw swelling measurement. Micro-CT imaging of excised rat hind paws (n = 10) stored in formalin consisted of approximately 600 30-mum slices acquired on a 512 x 512 image matrix with isotropic resolution. Following imaging, the joints were scored from H&E stained sections for cartilage/bone erosion, pannus development, inflammation, and synovial hyperplasia. From micro-CT images, quantitative analysis of absolute bone volumes and bone roughness was performed. Bone erosion in the rat AA model is substantial, leading to a significant decline in tarsal volume (27%). The result of the custom bone roughness measurement indicated a 55% increase in surface roughness. Histological and paw volume analyses also demonstrated severe arthritic disease as compared to controls. Statistical analyses indicate correlations among bone volume, roughness, histology, and paw volume. These data demonstrate that the destructive progression of disease in a rat AA model can be quantified using 3-D micro-CT image analysis, which allows assessment of arthritic disease status and efficacy of experimental therapeutic agents. PMID- 15802048 TI - Loss of p53 function in colon cancer cells results in increased phosphocholine and total choline. AB - Mutations in the p53 gene are the most frequently observed genetic lesions in human cancers. Human cancers that contain a p53 mutation are more aggressive, more apt to metastasize, and more often fatal. p53 controls numerous downstream targets that can influence various outcomes such as apoptosis, growth arrest, and DNA repair. Based on previous observations using (1)H magnetic resonance spectroscopy (MRS), we have identified choline phospholipid metabolite intensities typical of increased malignancy. Here we have used (1)H MRS to characterize the choline phospholipid metabolite levels of p53(+/ +) and p53(-/-) cells, and demonstrated that loss of p53 function results in increased phosphocholine and total choline. These data suggest that the increased malignancy of cancer cells resulting from loss of p53 may be mediated, in part, through the choline phospholipid pathway. PMID- 15802049 TI - Thermostability of firefly luciferases affects efficiency of detection by in vivo bioluminescence. AB - Luciferase from the North American firefly (Photinis pyralis) is a useful reporter gene in vivo, allowing noninvasive imaging of tumor growth, metastasis, gene transfer, drug treatment, and gene expression. Luciferase is heat labile with an in vitro halflife of approximately 3 min at 37 degrees C. We have characterized wild type and six thermostabilized mutant luciferases. In vitro, mutants showed half-lives between 2- and 25-fold higher than wild type. Luciferase transfected mammalian cells were used to determine in vivo half-lives following cycloheximide inhibition of de novo protein synthesis. This showed increased in vivo thermostability in both wild-type and mutant luciferases. This may be due to a variety of factors, including chaperone activity, as steady-state luciferase levels were reduced by geldanamycin, an Hsp90 inhibitor. Mice inoculated with tumor cells stably transfected with mutant or wild-type luciferases were imaged. Increased light production and sensitivity were observed in the tumors bearing thermostable luciferase. Thermostable proteins increase imaging sensitivity. Presumably, as more active protein accumulates, detection is possible from a smaller number of mutant transfected cells compared to wild-type transfected cells. PMID- 15802050 TI - Factors influencing quantification of in vivo bioluminescence imaging: application to assessment of pancreatic islet transplants. AB - The aim of this study is to determine and characterize factors influencing in vivo bioluminescence imaging (BLI) and apply them to the specific application of imaging transplanted pancreatic islets. Noninvasive quantitative assessment of transplanted pancreatic islets poses a formidable challenge. Murine pancreatic islets expressing firefly luciferase were transplanted under the renal capsule or into the portal vein of nonobese diabetic-severe combined immunodeficiency mice and the bioluminescence was quantified with a cooled charge coupled device camera and digital photon image analysis. The important, but often neglected, effects of wound healing, mouse positioning, and transplantation site on bioluminescence measurements were investigated by imaging a constant emission, isotropic light emitting bead (lambda = 600) implanted at the renal or hepatic site. The renal beads emitted nearly four times more light than hepatic beads with a smaller spot size, indicating that light absorption and scatter are greatly influenced by the transplant site and must be accounted for in BLI measurements. Detected luminescence decreased with increasing angle between the mouse surface normal and optical axis. By defining imaging parameters such as postsurgical effects, animal positioning, and light attenuation as a function of transplant site, this study develops BLI as a useful imaging modality for quantitative assessment of islets post-transplantation. PMID- 15802051 TI - Near-infrared optical imaging of integrin alphavbeta3 in human tumor xenografts. AB - In vivo optical imaging is potentially useful for evaluating the presence of tumor markers that are targets of molecular medicine. Here we report the synthesis and characterization of integrin alphavbeta3-targeted peptide cyclo(Lys Arg-Gly-Asp-Phe) [c(KRGDf )] labeled with fluorescence dyes with wavelength spanning from the visible/near infrared (Cy5.5) to the true near infrared (IRDye800) for optical imaging. In vitro, the peptide-dye conjugates bound specifically to tumor cells expressing alphavbeta3. When administered intravenously into mice at a dose of 6 nmol /mouse, the conjugates accumulated in tumors expressing alphavbeta3. The tumor-to-background ratios for human KS1767 Kaposi's sarcoma in mice injected with Cy5.5-c(KRGDf ) and Cy5.5 were 5.5 and 1.5, respectively. Preinjection of c(KRGDf ) blocked the uptake of Cy5.5-c(KRGDf ) in tumors by 89%. In alphavbeta3-positive M21 and alphavbeta3-negative M21-L human melanoma, fluorescence intensity in the tumor of mice injected with IRDye800 - c(KRGDf ) was 2.3 and 1.3 times that in normal tissue, respectively. Dynamic imaging revealed that Cy5.5- c(KRGDf ) was rapidly taken up by KS1767 tumor immediately after bolus injection. The rate of its uptake in the tumor was reduced by preinjection of c(KRGDf ) in an interval time-dependent manner. Our data suggest that near-infrared fluorescence imaging may be applied to the detection of tumors expressing integrin alphavbeta3 and to the assessment of the optimal biological dose and schedule of targeted therapies. PMID- 15802052 TI - Combined in vivo bioluminescence and fluorescence imaging for cancer gene therapy. AB - Nasopharyngeal carcinoma is intimately associated with the Epstein-Barr virus (EBV), which we have exploited therapeutically by constructing an EBV-specific synthetic enhancer sequence, within an adenoviral vector, denoted as adv.oriP. The achievement of tumor targeting provides therapeutic potential when delivered systemically, which could impact on distant metastases. We demonstrate here the feasibility and potential utility of combined, minimally invasive in vivo bioluminescence and fluorescence imaging to monitor adenoviral infection of subcutaneous C666-1 nasopharyngeal xenograft tumors stably expressing the DsRed2 gene. Fluorescence imaging was used to monitor the location and size of the C6661.DsRed2 tumors, whereas bioluminescence imaging demonstrated the distribution and specificity of a transcriptionally targeted adenoviral vector, adv.oriP.fluc, expressing the firefly luciferase gene. Fluorescence, bioluminescence, and photographic images were aligned using grids to examine colocalization of adenovirus and tumors. Bioluminescence and fluorescence co localized in 92% (11/12) of tumors at 24 hr and 100% (12/12) at 96 hr after adv.oriP.fluc (10(9) ifu) was administered intravenously. Nonspecific luciferase signal was detected in the liver area. The combined imaging was therefore successful in monitoring the uptake of systemically administered adenovirus in implanted tumors. This may ultimately lead to an effective noninvasive method to monitor the response of metastases to adenoviral gene therapy. PMID- 15802053 TI - Imaging gene expression in regional brain ischemia in vivo with a targeted [111in]-antisense radiopharmaceutical. AB - The gene encoding glial fibrillary acidic protein (GFAP) is downregulated 24 hr after reversible brain ischemia, such as with a middle cerebral artery occlusion (MCAO). The in vivo imaging of decreased GFAP gene expression in cerebral ischemia was examined in the present studies using a targeted peptide nucleic acid (PNA), which was labeled with (111)In, and which hybridized to nucleotides 20-37 of the rat GFAP mRNA. The PNA was monobiotinylated, and was attached to a monoclonal antibody (MAb) to the transferrin receptor (TfR) via a biotin streptavidin linkage. The TfR MAb enables trans-membrane transport of the PNA antisense radiopharmaceutical from blood to the cytosol of brain cells. The decreased GFAP gene expression at 24 hr after a 1-hr reversible MCAO was confirmed by immunocytochemistry. The [(111)In]-labeled PNA - MAb conjugate was administered intravenously to anesthetized rats at 24 hr after the 1-hr reversible MCAO, and the brain uptake of the targeted antisense imaging agent was decreased relative to brain regions outside of the infarct zone. These studies provide evidence that decreased expression of a target gene in brain can be imaged in vivo with a sequence-specific PNA, provided the antisense radiopharmaceutical is delivered across cell membranes with a receptor-specific targeting agent. PMID- 15802054 TI - Nanotechnology and tumor imaging: seizing an opportunity. AB - Nanoparticles, labeled with a signaling moiety for in vivo imaging, and one or more ligands for molecularly targeted specificity, hold considerable promise in oncology. Nanoparticles can serve as modular platforms, from which a wide variety of highly sensitive and specific imaging agents can be created. For example, many hundreds or thousands of atoms that provide imaging signals, such as radioisotopes, lanthanides, or fluorophores, can be attached to each nanoparticle, to form imaging agents that would provide higher sensitivity that can be obtained from agents based on small molecules. Similarly, many copies of targeted ligands can be attached to nanoparticles to markedly increase specific binding. Drugs or therapeutic isotopes can be added to create multifunctional nanoparticles. Appropriately labeled and targeted nanoparticles could lead to a paradigm change in which cancer detection, diagnosis, and therapy are combined in a single molecular complex. PMID- 15802055 TI - Nutritional deficiencies following bariatric surgery: what have we learned? AB - Deficiencies in vitamins and other nutrients are common following the Roux-en-Y gastric bypass (RYGBP), biliopancreatic diversion (BPD) and biliopancreatic diversion with duodenal switch (BPDDS), and may become clinically significant if not recognized and treated with supplementation. This paper presents a review of the current literature and evidence of the most commonly deficient vitamins and minerals following weight loss surgery, including protein, iron, vitamin B12, folate, calcium, the fat-soluble vitamins (A, D, E, K), and other micronutrients. The deficiencies appear to be more substantial following malabsorptive procedures such as BPD, but occur with restrictive procedures as well. The review suggests that further studies are needed to evaluate the clinical significance of the nutritional deficiencies, and to determine guidelines for supplementation. PMID- 15802057 TI - A comparison of burst pressure between buttressed versus non-buttressed staple lines in an animal model. AB - BACKGROUND: The consequences of a staple-line leak or disruption can be devastating particularly in the bariatric surgery population. This study was designed to assess if gastrointestinal staple-lines buttressed with bovine pericardium could tolerate greater intraluminal pressures compared with non reinforced staple-lines. METHODS: A laparoscopic linear cutting staple device was used to create divided staple-lines across the small intestine in a porcine model and the stomach in a rabbit model. 21 staple-lines were created with buttressing strips of bovine pericardium, and another 21 staple-lines were created without any reinforcement. All staple-lines were subjected to a constant rate increase in intraluminal pressure via intraluminal infusion of a dye solution. At the first sign of seam leak or failure, the burst pressure was recorded. RESULTS: In 19 of the 21 studies, the staple-lines buttressed with bovine pericardium sustained higher mean burst pressures than the conventional non-buttressed staple-lines. For the porcine intestinal segments, the mean intraluminal pressure at failure was 125 +/- 24 mmHg for buttressed staple-lines versus 58.4 +/- 28 mm Hg for conventional staple-lines (P <0.0001). For rabbit stomach segments, the mean intraluminal pressure at failure was also significantly higher for the buttressed segments (115 +/- 24 vs 75.6 +/- 23, P <0.0137). Most buttressed segments failed away from the staple-line, while the non-buttressed segments failed at the staple line. CONCLUSIONS: Gastrointestinal staple-lines buttressed with bovine pericardium are able to maintain seam integrity at significantly higher intraluminal pressures when compared to non-buttressed GI staple-lines in animal models. PMID- 15802056 TI - Long-term outcomes after gastric bypass. AB - BACKGROUND: While a great deal has been published regarding the short- and medium term outcomes of gastric bypass surgery, much less information is available regarding long-term follow-up. Such information would be valuable in changing attitudes towards this surgery. METHODS: 342 severely obese patients underwent gastric bypass between June 1990 and April 2003 by a single surgeon. Careful preoperative documentation and follow-up have been maintained on a computerized database. Where necessary, recent followup information has been gained by mailed questionnaire and blood tests. RESULTS: Follow-up data from within the last 12 months is available for 88% of patients. Follow-up time ranges from 0-14 years, with a median of 48.6 months. Of those lost to follow-up, only 24 (7%) have <12 months follow-up. The series includes 261 females and 81 males. Preoperative BMI ranged from 28-99 (median 44). Before surgery, hypertension was present in 138, type 2 diabetes in 62, and dyslipidemia in 265. There was no 30-day peri operative mortality. Three life-threatening complications occurred. BMI and % excess weight loss after 1, 2, 5 and 10 years were 28.7 and 89%, 28.3 and 87%, 31.2 and 70% and 31 and 75%, respectively. At most recent follow-up, 62% of those with hypertension before surgery were cured and 25% had improved. 85% of those with type 2 diabetes were cured and 10% had improved. No patients with impaired glucose tolerance had progressed to diabetes. 34% of those with dyslipidemia were cured and 38% had improved. CONCLUSION: The excellent outcomes, in terms of weight loss and improvement in co-morbidities, seen in both the short- and medium term after gastric bypass, are well maintained into the longer-term. PMID- 15802058 TI - Weight loss composition: the effects of exercise following obesity surgery as measured by bioelectrical impedance analysis. AB - BACKGROUND: Sudden weight loss following bariatric operations for morbid obesity, such as the duodenal switch (DS), can result in a concurrent decrease in lean body mass. Several methods for tracking body composition, such as bioelectrical impedance analysis (BIA), are available to monitor these changes. One method to offset the negative effects of sudden weight loss on body mass composition may be exercise. METHODS: 100 patients who had undergone the DS operation for morbid obesity were classified as exercisers and non-exercisers based on self-reporting. Their body mass compositions were measured using BIA preoperatively and at 0.75, 1.5, 3, 6, 9, 12, and 18 months postoperatively. RESULTS: At no study interval did postoperative percent changes in weight loss differ between the exercise and non-exercise groups. At 18 months postoperatively, the exercise group showed a 28% higher loss of fat mass and an 8% higher gain in lean body mass than the non exercise group. CONCLUSION: Exercise positively influences body mass composition following the DS. BIA can be successfully employed to monitor changes, diagnose deficiencies, and formulate treatment recommendations. PMID- 15802059 TI - Fasting plasma ghrelin levels increase progressively after biliopancreatic diversion: one-year follow-up. AB - BACKGROUND: The role of ghrelin in weight control after surgery is not clear. We examined plasma ghrelin and leptin levels in patients with morbid obesity undergoing biliopancreatic diversion (BPD) of Scopinaro. METHODS: 30 adult patients (27 females, 3 males), undergoing elective BPD were recruited from the Hospital Surgery Service. Fasting blood samples for biochemical determinations were drawn before surgery and 1, 3 and 12 months after BPD. Human plasma ghrelin was measured by RIA. RESULTS: During the study period, weight, BMI and serum leptin levels decreased significantly at all sample points compared to preoperative values. Ghrelin plasma levels increased during the study, with statistical significance at 3 months and 1 year after surgery compared with preoperative levels. While leptin changes correlated with changes in BMI, no correlation was found between ghrelin and leptin or BMI changes. CONCLUSION: Plasma ghrelin levels could be decreased in obese patients as a compensatory mechanism to their nutritional state, but our results do not support the postulated beneficial role of ghrelin in the 1-year weight loss after BPD. They rather suggest that weight loss somehow stimulates ghrelin secretion, even in the absence of part of the stomach. PMID- 15802060 TI - Weight changes in spouses of gastric bypass patients. AB - BACKGROUND: The effectiveness of gastric bypass for weight loss has been demonstrated. No study has documented the effect of this surgery on patient spouses' weight. Excess leftover food may be consumed by the spouses of gastric bypass patients, which may increase the total caloric intake compared to the preoperative intake. This investigation tested the hypothesis that there would be a significant change in patient spouse's weight. METHODS: Patients who underwent Roux-en-Y gastric bypass (RYGBP) > or = 12 months before were included. Preoperative weights of their spouses were requested before RYGBP surgery. Current weights of spouses were collected > or = 12 months postoperatively. Chi squared tests were used for statistical analysis. RESULTS: 63 patients were included in this study. 1 patient was divorced and current data was not available. 3 patients' spouses themselves had RYGBP during the study period, and their average weight loss was 49 kg. Of the spouses who had a significant change in weight (but did not have a RYGBP), 13 lost weight and 20 gained weight. In this subgroup of spouses, 75% (15/20) of obese spouses gained weight compared to only 38% (5/13) non-obese spouses (P <0.04). CONCLUSIONS: Patients' spouses who are obese are more likely to have weight gain while the patients lose weight after RYGBP. Preoperative counseling should be offered to spouses of potential bariatric surgery patients, especially to spouses who are obese. This phenomenon suggests that obese spouses of bariatric patients should be counselled regarding weight gain or even be considered for candidates for bariatric surgery. PMID- 15802061 TI - Weight loss and postoperative complications in morbidly obese patients with binge eating disorder treated by laparoscopic adjustable gastric banding. AB - BACKGROUND: The authors investigated the outcome of morbidly obese patients with binge eating disorder (BED) treated surgically with laparoscopic adjustable gastric banding. METHODS: The 5-year outcomes of 130 patients with BED and 249 patients without BED are described. The diagnosis of BED was made preoperatively and all patients with BED were supported with psychological therapy. RESULTS: Patients with and without BED had similar BMI levels before surgery. More patients with than without BED had depressive symptoms and associated minor disturbances of eating behavior (night eating and grazing). Percent excess weight loss (%EWL) in the first 5 years after surgery was similar in patients with and without BED. The percentage of BED patients showing %EWL >50% at the 5-year evaluation was 23.1, and 25.7% in non-BED patients. The percentage of patients showing weight regain in the last 4 years of follow-up was similar in binge eaters (20.8%) and in non-binge eaters (22.5%). The 5-year frequency of gastric pouch and esophageal dilatation was significantly higher in binge eaters than in non-binge eaters (25.4 vs 17.7 %, P<0.05 and 10.0 vs 4.8%, P<0.05, respectively). Binge eaters underwent a higher number of postoperative band adjustments than non binge eaters (3.0+/-2.1 vs 2.6+/-1.9, P<0.05) and the maximum band fill after surgery was higher in the BED patients than in non-BED patients (3.2+/-1.2 vs 2.8+/-1.3 ml, P<0.01). CONCLUSION: Morbidly obese patients with BED supported by adequate psychological treatment can have good outcomes after gastric banding. PMID- 15802062 TI - Factors influencing patient choice for bariatric operation. AB - BACKGROUND: No study has surveyed the factors that influence morbidly obese patients' preference for a particular bariatric operation. METHOD: 469 consecutive patients in 2 major bariatric surgery centers in the United States (US, 124) and Australia (AU, 345) were prospectively studied to determine referral pattern and reason for their choice of operation. RESULTS: The predominant operation was laparoscopic adjustable gastric banding (LAGB) in both US (75%) and AU (83%) centers. Gender (70% female), BMI (45 kg/m2) and age (42.5 years) were similar in both cohorts. In Australia, 53% had referral initiated by primary doctors and 25% by another patient, while in the US, 43% by another patient and 27% by the Internet. Safety of the operation (43%) was the highest rated factor in choosing LAGB. LAGB being "least invasive" was most significant in the US (46%), and "surgical safety" in Australia (45%). In the US, Rouxen-Y gastric bypass was preferred due to "lack of a foreign body" (31%) and "inability to cheat" (28%), while in Australia, "dumping" was the most significant reason (50%). Duodenal switch (BPD/DS) was selected in 11% of patients, primarily because of "durability of the weight loss" (51%). Surprisingly, only 1 patient in the US group selected BPD/DS because the pylorus remains intact. CONCLUSION: Safety and invasiveness had the greatest impact on patient choice for bariatric operation in two different countries. This information may help clinicians better understand their patients' concerns, and their treatment choices. PMID- 15802063 TI - Critical extreme anterior slippage (paragastric Richter's hernia) of the stomach after laparoscopic adjustable gastric banding: early recognition and prevention of gastric strangulation. AB - BACKGROUND: We report an unusual complication after Lap-Band placement characterized by herniation of the anterior stomach through the band. METHODS: Group 1 - 105 patients: Operated elsewhere (prior to US FDA approval) and followed by us; perigastric technique was used in 74 and pars flaccida in 31. Group 2 - 218 patients: Operated by us since August 2001 using the pars flaccida approach only. 4 patients with this unusual problem were identified. RESULTS: Patients were all female, with age 37.5 (3343) yr, weight 143.7 (123-167) kg, and BMI 54 (45-65). Onset occurred at 9 (5-16) months, with weight loss: 38.5 (27-53) kg and %EWL 47.3 (31-54)%. All had sudden nausea, vomiting and epigastric abdominal pain that persisted despite emptying the band. None of these symptoms were related to a recent band adjustment. CT scan showed a paragastric Richter's hernia of the stomach underneath the band. At exploration, the band was in the normal location. 3 patients from Group 2 had Richter's hernia of the anterior stomach through the band; reduction of the stomach with closure of the defect was performed. One patient from group 1 had gangrene of the entrapped stomach, resulting in band removal and gastrectomy. CONCLUSION: Lap-Band patients with sudden nausea, vomiting and abdominal pain, when not relieved by emptying the band, should undergo a CT scan. If a traditional slippage is not confirmed, paragastric Richter's hernia of the stomach through the band should be suspected. Immediate exploration with reduction of the stomach and closure of the defect can salvage the stomach and the band. Gastro-gastric sutures must completely close the space underneath the band to prevent this complication. PMID- 15802064 TI - Salvage of gastric restriction following staple-line dehiscence after vertical banded gastroplasty by insertion of an adjustable gastric band. AB - BACKGROUND: Vertical banded gastroplasty (VBG) has been a common and safe surgical treatment for morbid obesity. However, the complication of staple-line dehiscence (SLD) results in VBG failure. We present a minimally invasive revision procedure when SLD occurs: gastric restriction is salvaged by adjustable gastric banding (AGB), usually laparoscopically, providing that the previous restriction had achieved sufficient weight loss initially and was well-tolerated. METHODS: 13 patients with unexplained weight regain after VBG were found to have SLD on endoscopy. AGB was performed to re-establish restriction. Weight loss and complications were compared with two control groups: the first undergoing uncomplicated VBG, and the second undergoing AGB alone. Follow-up of 4.3+/-0.1 (mean+/-SEM) years after salvage reoperation, including complications, reoperations and weight loss, were studied. RESULTS: Insertion of the band through the retrogastric tunnel was feasible in all cases, despite adhesions in the area of the VBG Marlex band, the proximal stomach, and left lobe of liver. There was no surgeryrelated mortality. Following "salvage AGB", weight loss and overall complication rates were similar between the study group and the two control groups. CONCLUSION: Salvage of gastric restriction by AGB after SLD secondary to VBG is safe and reliable, despite the possibility of adhesions. Morbidity is low and intermediate-term weight loss is comparable to patients with uncomplicated VBG. PMID- 15802065 TI - Vertical banded gastroplasty: 6 years experience at a center in Poland. AB - BACKGROUND: Vertical banded gastroplasty (VBG) has been performed in our department as a restrictive operation for treatment of morbid obesity. We assessed efficacy, safety, and quality of life (QoL) after VBG, based on our 6 year experience. METHODS: 101 patients with >1 year follow-up who underwent VBG between January 1998 and May 2003, were retrospectively studied. Mean age was 41 years, and mean preoperative BMI was 51.3 kg/m2. Almost 60% of the patients suffered from hypertension, and 25% were diabetic. Postoperative QoL was determined with the BAROS questionnaire. Data concerning weight loss and co morbidities were collected during the postoperative visits. RESULTS: 87.2% of patients achieved > or = 50% EWL. 86 patients (85.2%) responded to the BAROS questionnaire. >90% of the patients analyzed according to BAROS, reported improvement in QoL after VBG. No patient reported deterioration in health or well being after the VBG. Anti-hypertensive medication was discontinued in 26 patients (56.5%) and decreased in the other 15 hypertensive patients (32.6%). 35% of diabetic patients did not require further treatment, while a further 40% had their insulin doses decreased or were switched to oral drugs. Early complications occurred in 4.65%, and consisted of an evisceration, pulmonary embolus and gastric leak. Late complications occurred in 20.9%, and included bleeding from peptic ulcer, incisional hernia, stomal stenosis and staple-line disruption (3.5%). There have been no deaths. CONCLUSION: VBG provided significant weight reduction and improved QoL in the vast majority of morbidly obese patients. Patients with diabetes and hypertension benefitted because these co-morbidities were improved or disappeared with the weight loss. PMID- 15802066 TI - Histological changes in the liver of morbidly obese patients: correlation with metabolic parameters. AB - BACKGROUND: Obesity is a major risk factor for fatty liver disease. The purpose of this study was: 1) to determine the degree of steatosis, inflammation and fibrosis in liver biopsies of morbidly obese patients in relation to their body fat distribution and metabolic status, and 2) to examine the course of liver enzyme changes with surgically-induced weight loss. METHODS: The study population included 179 morbidly obese bariatric surgical patients (82% female, 18% male, mean age 39+/-0.7 (SEM) years, BMI 52+/-0.6 kg/m2, excess body weight 80+/-1.8 kg). All patients tested negative for hepatitis and HIV. Liver biopsies were taken intra-operatively. Hepatic enzyme activities were measured along with lipid parameters, fasting glucose, insulin and leptin. RESULTS: Liver biopsies showed that 47% of morbidly obese females and 85% of males had >30% of hepatocytes filled with fat droplets. Clinically significant hepatic steatosis was associated (P<0.01) with: a) metabolic aberrations, i.e.hyperlipidemia, hyperglycemia, b) male gender, c) abdominal adiposity, and d) elevated hepatic aminotransferase activities. Hepatic inflammation was found in 47% of females and 55% of males, and 'moderate' fibrosis occurred in 12% of males and 6% of females. Postoperatively, the activity of hepatic aminotransferases declined after an initial increase in response to weight loss, with normalization of values occurring at an excess weight loss of 50% (P<0.0001). CONCLUSION: The majority of morbidly obese patients have >30% steatosis of the liver. The incidence of steatosis is higher for males than females, possibly due to their visceral obesity and associated metabolic aberrations. PMID- 15802067 TI - Gallbladder pathology in morbid obesity. AB - BACKGROUND: Obese patients are at increased risk for biliary disease. The prevalence and type of gallbladder pathology in morbidly obese patients was evaluated, and compared with a non-obese control group. METHODS: A consecutive series of obese patients (n=478) who had undergone bariatric surgery with concurrent routine cholecystectomy and a consecutive group of organ donors (n=481) were compared. Gallbladder pathology was defined as: cholelithiasis, cholecystitis, cholesterolosis, or normal pathology. RESULTS: Mean age of obese patients and of donors was 42 +/- 9 and 43 +/- 17 years respectively and mean BMI was 52 +/- 10 and 27 +/- 7 kg/m2 respectively, P<0.05. There were more females in the obesity group (88% vs 47%, P<0.0001). 31% of obese patients and 7% of controls had a previous cholecystectomy (P<0.0001). 21% of the obese and 72% of the controls had normal gallbladder pathology (P<0.0001). Overall, obese patients had a higher incidence of cholelithiasis (25% vs 5%, P<0.0001), cholecystitis (50% vs 17%, P<0.0001), and cholesterolosis (38% vs 6%, P<0.0001) compared with controls. Obese patients with BMI <50 were more likely than those with BMI > or =50 to have normal gallbladder pathology (27% vs 14%, P<0.001). Female patients were more likely to have undergone previous cholecystectomy than males in both the obese group (34% vs 11%, P<0.001) and the control group (12% vs 2%, P<0.0001). Normal pathology was more common in male patients (80% vs 63%, P<0.0001) and patients <50 years (76% vs 66%, P<0.05) in the control group. CONCLUSIONS: Obese patients have an increased incidence of benign gallbladder disease than a group of controls, and the relative risk appears to be positively correlated with the level of increase in the BMI. Obesity appears to change the effect of age and gender on gallbladder pathology. PMID- 15802068 TI - Laparoscopic cholecystectomy in obese patients. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) is the treatment of choice for gallstones. Obesity was initially considered a contraindication to this approach. The aim of this report is to review our experience with LC, to evaluate the role of BMI in the outcome. METHODS: The records of 1,804 patients who underwent LC for symptomatic cholelithiasis from May 1992 to January 2004 were analyzed retrospectively. Patients were divided into 5 groups according to their BMI: < or =24.9, 25.0-29.9, 30.0-34.9, 35.0-39.9 and > or =40 kg/m2. RESULTS: Of the 1,804 patients [1,379 females (76.4%) and 425 males (23.6%)] who underwent LC, 431(23.9%), 924 (51.2%), 355 (19.7%), 68 (3.8%) and 26 (1.4%) had BMI values of < or =24.9, 25.0-29.9, 30.0-34.9, 35.0-39.9 and > or =40 kg/m2, respectively. Conversion to open cholecystectomy was required in 94 patients (5.2%), and complications occurred in 39 patients (2.2%). There was no correlation between BMI and the conversion rate (P=0.593) and complication rate (P=0.944), while the hospital stay was similar between the groups with successful LC. The only significant difference was the longer operating time in the two obesity groups (P<0.001). CONCLUSIONS: LC is effective and safe in patients with morbid obesity. As it carried low risks of conversion and perioperative complications, we suggest that LC is the select approach for these patients. Moreover, the rapid mobilization and hospital discharge following LC may provide extra benefit to these patients. PMID- 15802069 TI - Visceral sensitivity to gastric stimulation and its correlation with alterations in gastric emptying and accommodation in humans. AB - BACKGROUND: The aim of this study was to investigate the visceral sensation to gastric stimulation and its correlation with the stimulation-induced alterations in gastric accommodation and gastric emptying. METHODS: The study was performed in 12 healthy human controls. Gastric stimulation was performed using bipolar electrodes attached to the mucosa of the distal stomach under endoscopy. Experiments were conducted on 3 consecutive days to investigate the effects of gastric stimulation with various parameters on visceral sensation, maximum intake of water and gastric emptying of solids. RESULTS: 1) The stimulation energy was 265.6 +/- 134.9 smA 2 for the first sensation and 2,020.0 +/- 865.1 smA2 for the maximum tolerance, and there was a large variation among the subjects; 2) Gastric stimulation with stimulation energy less than 50% of that required to induce the first sensation significantly reduced the maximum intake of water and delayed gastric emptying of solids without inducing symptoms; 3) The stimulation energy of the first sensation was significantly correlated with the stimulation-induced reduction in water intake (r=-0.80, P=0.02) and the stimulation-induced prolongation of gastric emptying (r=-0.78, P=0.003). That is, the inhibitory effects of gastric stimulation were more obvious in those subjects who were viscerally more sensitive to gastric stimulation. CONCLUSIONS: Gastric stimulation via the distal stomach reduces gastric accommodation and delays gastric emptying. These inhibitory effects are correlated with the visceral sensitivity of the individual to gastric stimulation. It is worthy to further investigate whether the outcome of the implantable gastric stimulation (IGS) therapy for obesity may be predicted from the visceral sensitivity of the patient to temporary gastric stimulation using endoscopically placed mucosal electrodes. PMID- 15802070 TI - Efficacy of the SF-36 questionnaire in identifying obese patients with psychological discomfort. AB - BACKGROUND: The primary endpoint of this study was to assess the association of health-related quality of life (QoL) and the presence of psychopathology. The association of other patients' characteristics and of Cognitive Behavioral Assessment (CBA) scales with quality of life (QoL) was also evaluated. METHODS: 100 consecutive obese patients (WHO grade 2 and 3 obesity), addressed for psychological advice before either invasive or non-invasive treatment of obesity, were investigated. The instruments used were the SF-36 questionnaire (physical and mental component summaries, PCS and MCS), the CBA scales and psychological counselling. The association of PCS and MCS with the presence of psychopathology (Marked or DSM IV discomfort) was assessed by means of logistic regression. RESULTS: SF-36 PCS was 39.5 (95% CI 37.7-41.3) and MCS 49.8 (95% CI 47.7-51.9). PCS only was significantly lower than the average for the reference normal population. The mean PCS score was similar in the No-Moderate (39.6 (SD 7.6)) and Marked-DSM IV (39.1 (SD 7.6)) groups, with an adjusted odds ratios (OR) of 1.07 (95% CI 0.74-1.55), P=0.706, for 5 points increase in PCS. The mean MCS score was 51.7 (SD 10.3) in the No-Moderate group and 42 (SD 8.1) in the Marked-DSM IV group, with an adjusted OR for 5 points increase in score of 0.63 (95% CI 0.43 0.95), P=0.003. CONCLUSIONS: SF-36, and particularly the MCS component, is a simple tool of easy use that could be utilized for identifying patients needing a specific psychological intervention in severely obese subjects applying for a weight reduction program. PMID- 15802071 TI - Compliance with surgical after-care following bariatric surgery for morbid obesity: a retrospective study. AB - BACKGROUND: Non-compliant patients fail to match their behavior to the clinical prescription. Laparoscopic adjustable gastric banding requires strict compliance with surgical and dietary advice. Failure to attend follow-up appointments and the persistent consumption of calorie-dense liquid foods are associated with poor weight loss and postoperative complications. Prediction of "poor compliers" would enhance candidate selection and enable specific interventions to be targeted. METHODS: 9 poor compliers were identified and compared with 9 fully compliant controls. Case-notes were analyzed retrospectively. RESULTS: Cases were found to graze on foods and eat more in response to negative affects. They were reluctant to undergo psychiatric assessment, viewed the band as responsible for weight loss, and aroused caution in the psychiatric evaluator. Poor compliance was not associated with binge eating, purging, impulsivity or psychiatric illness. CONCLUSIONS: Unrealistic expectations and anxiety are known to predict non adherence. Constant negative affects may be self-modulated by grazing. The results are explored in the context of Self-efficacy Theory, a socio-cognitive account of illness behavior. PMID- 15802072 TI - Exploring the impact of obesity surgery on patients' health status: a quantitative and qualitative study. AB - BACKGROUND: Obesity surgery has been shown to result not only in sustained weight loss, but also in improvements in psychological morbidity and quality of life. The present study aimed to explore the mechanisms behind the success of surgery and to examine how it might bring about such changes in patients' health status. METHODS: Both quantitative and qualitative methods were used. A questionnaire was completed by patients who had undergone bariatric surgery in the past 4 years (n=22) and a matched-group of waiting list controls (n=39). In depth interviews were then carried out with 15 surgical patients. RESULTS: The quantitative study showed that the surgical group weighed less, had more negative experiences of eating and yet reported improved subjective health status using a range of validated tools. The qualitative study provided some insights into this improved health status, with the surgical patients reporting improved energy and self esteem. The patients also reported a fundamental shift in their relationship with food. In particular, they described changes in their eating behavior and stated that they had a new experience of fullness and a reduction in their hunger. Furthermore, they described how food now had a reduced role in their lives and that the operation had made them feel more in control of their food intake. CONCLUSION: Post-surgical improvements in health status may not only be a result of the non-specific consequences of surgery brought about by weight loss, but also a result of the specific impact of the surgical procedure itself. By enforcing a reduction in the amount of food that can be eaten, restrictive surgery seems to generate changes in the individuals' relationship to food and may help to re-establish a perception of control over eating behavior. PMID- 15802073 TI - Response of recurrent binge eating and weight gain to topiramate in patients with binge eating disorder after bariatric surgery. AB - BACKGROUND: The effectiveness of topiramate was evaluated in the treatment of recurrent binge eating and weight gain in patients with binge eating disorder (BED) and obesity who had undergone initially successful bariatric surgery. METHODS: The records of 3 consecutive patients with BED and obesity who presented to our clinic with recurrent binge eating and weight gain after undergoing initially successful bariatric surgery were reviewed. They were treated with topiramate for an average of 10 months. RESULTS: All three patients reported complete amelioration of their binge eating symptoms and displayed weight loss (31.7 kg in 17 months, 14.5 kg in 9 months, 2 kg in 4 months, respectively) in response to topiramate (mean dose 541 mg). CONCLUSION: Although anecdotal, these observations suggest that topiramate may be an effective treatment for patients with BED and obesity who experience recurrent binge eating and weight gain after initially successful bariatric surgery. PMID- 15802074 TI - Hepatic portal gas due to gastro-jejunal anastomotic leak after laparoscopic gastric bypass. AB - Gastrointestinal complications after laparoscopic Roux-en-Y gastric bypass (LRYGBP) are not uncommon. Leakage of the gastro-jejunal anastomosis is the main early surgical complication of LRYGBP. Hepatic portal venous gas (HPVG) has been described in association with a variety of pathologic conditions. HPVG is a potentially life-threatening condition due to its etiology, with a global survival rate of <25%. We present a case of gastro-jejunal anastomotic leak associated with HPVG after LRYGBP. PMID- 15802075 TI - Topical anesthesia-induced methemoglobinemia in bariatric surgery patients. AB - Methemoglobinemia leads to rapid oxygen desaturation, requiring prompt recognition and treatment. We present two severely obese patients who developed methemoglobinemia following the use of topical or local anesthetic. This complication was detected by analysis of arterial blood gases, and was successfully treated with methylene blue i.v. and 100% O2 supplementation. PMID- 15802076 TI - Biliopancreatic diversion as a novel approach to the HAIR-AN syndrome. AB - BACKGROUND: The HAIR-AN syndrome is a rare multisystem disorder in women, that consists of hyperandrogenism (HA), insulin resistance (IR) and acanthosis nigricans (AN). The IR is likely due to a primary defect of the insulin receptor. METHODS: We report the case of a 42-year-old Caucasian woman with HAIR-AN syndrome, impaired glucose tolerance (IGT), mild hyperlipemia, and hypertension, who underwent biliopancreatic diversion (BPD). RESULTS: Within 24 months follow up after BPD, impaired glucose tolerance, mild hyperlipemia, and hypertension completely reversed. Although insulin sensitivity, estimated by the euglycemic hyperinsulinemic clamp, did not improve, signs and symptoms of hyperandrogenism and acanthosis nigricans resolved fully. CONCLUSION: In HAIR-AN syndrome, malabsorptive bariatric surgery is effective in improving hyperandrogenism and acanthosis nigricans, with noteworthy esthetic consequences. BPD was followed by disappearance of co-morbidities of the syndrome, such as IGT, hypertension and hyperlipemia. PMID- 15802077 TI - The lethality of obstructing hematoma at the jejunojejunostomy following Roux-en Y gastric bypass. PMID- 15802078 TI - The lethality of obstructing hematoma at the jejunojejunostomy following Roux-en Y gastric bypass. PMID- 15802080 TI - Effect of gastric banding on ileal mucosa. PMID- 15802082 TI - The neurodevelopmental model of schizophrenia: what can very early onset cases tell us? PMID- 15802083 TI - The selective serotonin reuptake inhibitors controversy in the treatment of depression in children. AB - Antidepressants are widely prescribed for children and adolescents, although data regarding their safety and efficacy are limited. The objective of this article is to review the origins of the controversy regarding the current use of selective serotonin reuptake inhibitors (SSRIs) in children and adolescents. Two chief concerns drive the controversy: 1) the discovery of an increased risk of suicidal behaviors in those treated with SSRIs and 2) the efficacy of SSRIs in childhood and adolescent major depression is unclear. Various factors may account for the reported differences in outcomes for SSRI treatment in children and adolescents compared to adults. The past decade has shown a significant drop in the rate of adolescent suicide, which coincides with the onset of the use of these medications. Therefore, a reduction in the use of SSRIs in children and adolescents should be considered carefully. PMID- 15802084 TI - Treatment of mania in children and adolescents. AB - Bipolar disorder (BPD) is a severe and complex illness that seriously disrupts the lives of those afflicted. Increased rates of suicide attempts and completions, poorer academic performances, disturbed interpersonal relationships, increased rates of substance abuse, legal difficulties, and multiple hospitalizations all have been associated with BPD. At least 1% of children and adolescents have this disorder. This commentary is designed to review and summarize the recent literature on the treatment of manic and/or hypomanic phases of BPD in children and adolescents. PMID- 15802085 TI - How to differentiate bipolar disorder from attention deficit hyperactivity disorder and other common psychiatric disorders: a guide for clinicians. AB - Bipolar disorder in children often is confused with attention deficit disorder, substance-induced mood disorder, oppositional defiant disorder, and conduct disorder. It is not uncommon for some of these disorders to be comorbid with pediatric bipolar disorder. This article provides the reader with a review of the existing literature on differentiating these illnesses and recognizing the phenomenology of each disorder as it pertains to a psychiatric diagnostic work-up of a child. Clinically helpful overlapping and unique characteristics of each disorder are discussed and a practical approach to differentiate these disorders is provided. PMID- 15802087 TI - The impact of genetics on child psychiatry: a 20-year perspective. PMID- 15802086 TI - Prepubertal bipolar disorder: proper diagnosis should lead to better treatment response. AB - Treatment research in prepubertal bipolar disorder remains in a rudimentary stage. Phenomenological evidence suggests it is a heterogeneous disorder with varying degrees of rapid cycling, aggression, and psychosis often accompanied by comorbid diagnoses of attention deficit hyperactivity disorder, conduct disorder, oppositional defiant disorder, and anxiety disorders including obsessive compulsive disorder. Longitudinal and family history studies suggest prepubertal bipolar disorder may be more treatment-resistant than later-onset bipolar disorder. Neurobiological studies to guide treatment, though promising, remain in their infancy. Clinical trials to date (mostly open studies) often have lumped together subjects with manic, hypomanic, and mixed presentations with different and/or undiagnosed comorbidities, making meaningful comparisons of treatment response difficult. Randomized, double-blind, placebo-controlled trials are needed to clarify best treatment options for bipolar subtypes with and without comorbid disorders. More homogeneous diagnostic groupings based on episode and duration criteria and a more patient-centered, symptom-based approach should be considered in treatment designs. PMID- 15802088 TI - The genetics of depression and related traits. AB - There is considerable evidence that genetic factors play a major role in the etiology of unipolar depression. Investigations into vulnerability genes for unipolar depression are underway and for more broadly defined depression-related traits, such as anxiety, neuroticism, and harm avoidance. This review discusses some of the core issues related to study design and molecular genetic methodology, followed by an overview of recent molecular genetic findings for unipolar depression. The research to date has identified regions within certain chromosomes that may contain risk genes. Improved study design and the use of new molecular techniques hold promise for the identification of more specific vulnerability genes for unipolar depression. PMID- 15802090 TI - Genetic advances in autism hinge on the method of measuring symptoms. AB - Advances in genotyping and related technologies and the study of families with two or more autistic siblings have allowed the reporting of various genetic linkage and candidate gene association studies for autism. Despite this significant research effort, there still is no general agreement on the role of particular genes that predispose to the risk of autism. One source of general debate is whether core symptoms studied in research on the heredity of autism include single or multiple independent genetic domains. In this review, it is argued that failure to control for co-existing dimensions of psychopathology and lack of assessment of parents may lead to misinterpretation of genetic findings from pedigrees involving multiple affected family members. PMID- 15802089 TI - Combining research approaches to advance our understanding of drug addiction. AB - Drug addiction is a complex behavior, likely to be influenced by various genes, environmental factors, and gene-gene and gene-environment interactions. Various aspects of addiction are studied by different disciplines. Animal studies are increasing insight into brain regions and genes associated with addiction. Epidemiologic studies are establishing the factors increasing risk for initiation and continuation of substance use. Twin and adoption studies are increasing our understanding of the complex mechanisms involved in substance use, including comorbidity and gene environment interaction. Finally, molecular genetic studies in humans are starting to yield some converging findings. It is argued and illustrated with examples that greater awareness of progress in other disciplines can speed up our understanding of the complex processes involved in addiction. This should help our ability to identify who is at increased risk of becoming addicted and the development of prevention and intervention strategies targeted at an individual's specific needs. PMID- 15802091 TI - Ebrii gignunt ebrios--One drunkard begets another: the genetics of alcohol dependence. AB - Family, twin, and adoption studies provide convincing evidence of a genetic contribution to the predisposition to alcohol dependence. Two main molecular genetic approaches, namely linkage and association, have been adopted to identify the genes that underpin that genetic vulnerability. Robust findings have implicated genes involved in alcohol metabolism, particularly when studied in Asian populations. These include the genes encoding aldehyde dehydrogenase 2 and several alcohol dehydrogenases. Other findings have proven to be less resilient, but have often implicated genes of the dopaminergic, GABA-ergic and serotonergic systems. Improvements in sample collection and characterization, technological and statistical advances, combined with a developmental approach, should enable molecular genetics to deliver on its initial promise of identifying the genes involved. These findings will have important implications for the identification and targeting of treatment. However it is anticipated that this information will have little impact on risk alteration and consequently population screening will be highly unlikely. Genetic factors always should be considered in the context of their developmental interplay with those of the environment. PMID- 15802093 TI - Autoimmune hepatitis. PMID- 15802094 TI - Antibiotic prophylaxis for severe acute pancreatitis. PMID- 15802092 TI - Meta-analysis in psychiatric genetics. AB - The article reviews literature on methods for meta-analysis of genetic linkage and association studies, and summarizes and comments on specific meta-analysis findings for psychiatric disorders. The Genome Scan Meta-Analysis and Multiple Scan Probability methods assess the evidence for linkage across studies. Multiple Scan Probability analysis suggested linkage of two chromosomal regions (13q and 22q) to schizophrenia and bipolar disorder, whereas Genome Scan Meta-Analysis on a larger sample identified at least 10 schizophrenia linkage regions, but none for bipolar disorder. Meta-analyses of pooled ORs support association of schizophrenia to the Ser311Cys polymorphism in DRD2 and the T102C polymorphism in HTR2A, and of attention deficit hyperactivity disorder to the 48-bp repeat in DRD4. The 5-HTTLPR polymorphism in the serotonin transporter gene (SLC6A4) may contribute to the risk of bipolar disorder, suicidal behavior, and neuroticism, but association to the lifetime risk of major depression has not been shown. Meta analyses support linkage of schizophrenia to regions where replicable associations to candidate genes have been identified through positional cloning methods. There are additional supported regions where susceptibility genes are likely to be identified. Linkage meta-analysis has had less clear success for bipolar disorder based on a smaller dataset. Meta-analysis can guide the prioritization of regions for study, but proof of association requires biological confirmation of hypotheses about gene actions. Elucidation of causal mechanisms will require more comprehensive study of sequence variation in candidate genes, better statistical and meta-analytic methods to take all variation into account, and biological strategies for testing etiologic hypotheses. PMID- 15802095 TI - The epidemiology and impact of pancreatic diseases in the United States. AB - In the United States, acute pancreatitis, chronic pancreatitis, and pancreatic cancer are the most common pancreatic disorders requiring diagnosis and treatment. Pancreatic cancer is responsible for nearly 30,000 annual deaths and is the second most common cause of death from any type of gastrointestinal disease. Gallstone disease, which is strongly associated with obesity, and excessive consumption of alcohol are the major risk factors for benign pancreatic disease, whereas smoking is the most important factor known to cause pancreatic cancer. Therefore, to reduce the overall burden of pancreatic disease, we should focus on the control of three lifestyle factors: smoking, drinking, and obesity. PMID- 15802096 TI - Alcoholic pancreatitis: new insights into an old disease. AB - Alcoholic pancreatitis is an old disease that continues to present controversial issues. One of the most hotly debated issues is whether alcoholic pancreatitis is a chronic disease from the beginning or if instead it becomes chronic after repeated episodes of acute pancreatitis. Histologic studies, including very large series of patients with alcoholic pancreatitis, have clearly shown that this disease is chronic from the beginning and that, if acute necrotic pancreatitis occurs, it is associated with chronic lesions. The possibility that acute alcoholic pancreatitis can occur in the absence of chronic lesions cannot be excluded, but, if this occurs, it is rare. In addition to alcohol, genetic factors certainly play a determining role. Until now many genetic studies have been made on chronic pancreatitis; the first dealt with hereditary pancreatitis. In this disease it has been shown that mutations of the cationic trypsinogen gene and of SPINK1 are implicated in its pathogenesis. Concerning alcoholic pancreatitis, several studies have been made, but the results so far are disappointing. PMID- 15802097 TI - Autoimmune pancreatitis. AB - The list of diseases associated with autoantibodies against tissues, cells, or specific autoantigens is growing, and many organs in the body are known to be affected by autoimmune injury. Until recently, the most well-known pancreatic autoimmune disorder was type 1 diabetes mellitus, where there is selective destruction of beta cells in the islets of Langerhans. Although an autoimmune process affecting the exocrine pancreas was suspected over four decades ago, it is only in the past 10 years or so that autoimmune chronic pancreatitis has been recognized as a distinct entity. Here we review the clinical, serologic, radiologic, and histologic features of autoimmune pancreatitis. PMID- 15802098 TI - An endoscopic perspective on pancreatic pseudocysts. AB - Significant advances in endoscopic, radiologic, and surgical techniques have transformed the management of pancreatic pseudocysts. The present review focuses upon advances in the endoscopic management of pseudocysts and incorporation of these techniques into an overall management approach. Findings with endoscopic ultrasound (EUS) and endoscopic retrograde cholangiopancreatography often guide the choice of drainage method. Endoscopic drainage can be achieved through the transpapillary or transmural routes. EUS has increasingly become an integral part of transmural pseudocyst drainage. PMID- 15802099 TI - Advances in evaluation and management of gastrinoma in patients with Zollinger Ellison syndrome. AB - Gastrinomas producing Zollinger-Ellison syndrome are the most frequent symptomatic, malignant pancreatic endocrine tumor syndrome. Recently, a number of important studies have examined their molecular pathogenesis and natural history and provided important guidelines for their treatment. Each of these areas is briefly reviewed in this article. PMID- 15802100 TI - Risk factors for pancreatic adenocarcinoma: are we ready for screening and surveillance? AB - Of all common malignancies, pancreatic cancer has the lowest survival rate. According to estimates by the American Cancer Society, in 2004, 31,860 Americans were diagnosed, and 31,270 died from this near universally lethal disease. Unfortunately, only a small minority of patients are diagnosed when the extent of disease is still localized and thus potentially curable. These dismal survival results could be improved by developing successful screening and prevention strategies. Routine screening is not currently suggested for pancreatic cancer; however, there may be a benefit in a select group of patients identified to be at high risk for developing this disease. It is recommended that surveillance be performed on these high-risk individuals in a research setting to allow for the development of a successful detection strategy. PMID- 15802101 TI - Sphincter of Oddi manometry and post-ERCP acute pancreatitis. PMID- 15802103 TI - Is sphincter of Oddi manometry a risk factor for pancreatitis? A different view. AB - Studies suggest a causal relationship between sphincter of Oddi manometry (SOM) and acute pancreatitis, presumably due to water instillation in the ductal system. In this article, critical analysis of the existing studies attributes the high risk of acute pancreatitis with SOM to "association due to confounding" rather than to "causation." This conclusion is based on two pieces of evidence: The first is lack of biologic plausibility: Biologic evidence is lacking to support the hypothesis that water instillation during SOM can induce acute pancreatitis. The second is confounding evidence: Manometric studies show considerable variation in the risk of post-procedure pancreatitis (4% to 30%), which suggests that other important factors besides SOM influence the adverse outcome. These studies did not control for other variables, which are well known to predispose to pancreatitis and therefore may act as confounding factors. Two studies that assessed the independent role of SOM in causation of acute pancreatitis showed that SOM is not an independent predictor of acute pancreatitis. It is the underlying disorder sphincter of Oddi dysfunction, and not the "SOM" procedure, that predisposes patients to post-procedure acute pancreatitis. PMID- 15802102 TI - Epidemiology and risk factors for gallstone disease: has the paradigm changed in the 21st century? AB - Gallstone disease is common and costly, creating over 700,000 cholecystectomies annually. Its complications consume approximately $6.5 billion in the United States. Surveys using noninvasive ultrasonography have identified its true prevalence and the associated risk factors. In developed countries, at least 10% of white adults harbor cholesterol gallstones; women have twice the risk, and age further increases the prevalence in both sexes. Gallstones reach epidemic proportions in the North and South American Indian populations, accompanied by an increased risk for gallbladder cancer. In contrast, the rate in sub-Saharan Africa and Asia is quite low. Obesity, a major risk factor, likely relates to insulin resistance (the metabolic syndrome). Evolution and circumstance in American Indians may have ironically selected those with "thrifty" genes that conserve energy. Our abundant access to food places us at the increased risk of obesity and cholelithiasis. The general rise in obesity in many countries raises the specter of heightened disease, best identified by epidemiologic studies. PMID- 15802104 TI - The gallbladder and biliary tract in cystic fibrosis. AB - Chronic liver disease is a major complication of cystic fibrosis. Its incidence and severity are variable, and diagnosis relies on a combination of clinical evaluation, biochemical testing, and radiologic assessment. Identifying patients who have early disease is critical, and the administration of ursodeoxycholic acid appears to be beneficial. The pathogenesis is incompletely understood, and factors that contribute to the variability in incidence and severity are unknown. Fortunately, only a small proportion of individuals progress to advanced liver disease; however, in this population, there is significant morbidity and impairment in quality of life. Liver transplantation can be performed successfully in patients with end-stage liver disease. Future treatments involve targeted gene therapy and activation of mutant forms of the cystic fibrosis transmembrane conductance regulator. PMID- 15802105 TI - Biliary scintigraphy versus sphincter of Oddi manometry in patients with post cholecystectomy pain: is it time to disregard the scan? AB - Sphincter of Oddi (SO) dysfunction is diagnosed using manometry, and patients with an abnormal SO basal pressure respond well to division of the SO. However, manometry is invasive and is associated with a low, yet significant, incidence of complications. Scintigraphy techniques have been developed with the aim of providing a noninvasive means of assessing SO motility. However, when compared with SO manometry these techniques fall short in sensitivity and specificity for diagnosing SO dysfunction. Furthermore, they do not select patients who will respond to treatment. Consequently, the quest for development of a noninvasive investigation for diagnosis of SO dysfunction continues. In the mean time, improved manometric techniques that enhance reproducibility and reduce complications have been developed. PMID- 15802106 TI - Biliary complications in liver transplant recipients. AB - Despite numerous advancements in the management of patients who have undergone cadaveric liver transplantation, biliary complications continue to challenge clinicians. Biliary leaks in the early postoperative period and strictures in the late postoperative period represent the two major posttransplant biliary complications. Intrahepatic and hilar strictures are particularly difficult to manage and frequently require retransplantation, which should not be delayed. Choledocholithiasis and the biliary cast syndrome are frequently associated with underlying biliary strictures, and endoscopic attempts at removal should precede surgical interventions. Sphincter of Oddi dysfunction is increasingly recognized despite unclear pathophysiology. Hemobilia is most commonly iatrogenic and requires a high suspicion and prompt intervention. Although the number of diagnostic and therapeutic options have increased, there is no consensus as to which is superior. In recent years there has been a trend toward nonsurgical interventions, in particular endoscopic approaches. PMID- 15802108 TI - [New perspectives for the treatment of diabetes mellitus]. PMID- 15802109 TI - [Effectiveness of self-measurement of blood pressure in patients with hypertension: the Dioampa study]. AB - OBJECTIVE: To evaluate the effectiveness of self-measurement of blood pressure (SMBP) in controlling hypertension. DESIGN: Randomized, controlled, pragmatic, open study. The unit of randomization was the basic health care unit (BCU), consisting of 1 physician and 1 nurse. All BCUs were randomized to the control group (usual clinical practice, n=94) or to the intervention group (n=86). SETTING: Primary care BCUs throughout Spain. PARTICIPANTS: Patients with poorly controlled essential hypertension, defined as systolic blood pressure > or = 140 or diastolic blood pressure > or = 90 mm Hg. INTERVENTIONS: The patients were given an OMRON HEM-705CP automatic blood pressure monitor on two occasions, for use during 15 days at weeks 6 and 14. Blood pressure was recorded at each visit (baseline, 6, 8, 14, 16, and 24 weeks). Main outcome measures. MAIN OUTCOME VARIABLE: control of blood pressure, considered systolic/diastolic blood pressure <140/90 mm Hg (130/85 in patients with diabetes). RESULTS: 180 BCUs serving 1325 patients (622 in the intervention group, 703 in the control group) participated. Baseline characteristics were similar in both groups. Immediately after the first period of SMBP (week 8) the proportion of patients whose blood pressure was well controlled was 7.6% higher in the intervention group than in the control group (P=.01). After the second period of SMBP (week 16) the difference between groups decreased to 4.1% (P=.27). At the end of the study the difference was 4.9% (P=.19). CONCLUSIONS: Self-measurement of blood pressure was effective in controlling blood pressure in the short term, but its effects faded over time. PMID- 15802110 TI - [Self-measurement of blood pressure, therapeutic compliance and monitoring of blood pressure]. PMID- 15802111 TI - [The need for bioethics training in the Community of Madrid]. AB - OBJECTIVES: MAIN OBJECTIVE: to understand the significance and sense of bioethics for social work/health professionals; the secondary objective is to quantify the bioethics training given in IMSALUD primary care. DESIGN: Interviews with key reporters. Face-to-face interviews with people in charge of the training and research departments. SETTING: The 11 primary health care areas of IMSALUD. PARTICIPANTS AND/OR CONTEXTS: Key reporters (1 for each health area) chosen by heads of teaching: people working in the area with a specific weight or personal interest in bioethics. METHOD: 11 key reporters (1 for each health area); telephone contact and mailing of open-question questionnaire. RESULTS: Bioethics is implicit in daily activity, but unconsciously. General rejection due to its themes, which involve self-criticism. Ethical conflicts basically arising from the relationship of health workers with patients and the family, especially concerning information-giving, were pointed to. Themes considered very important were informed consent and confidentiality. Other ethical problems were allocation and distribution of limited resources, responsibility for taking up situations that worsen health care (lists too long, burnt-out companions, malpractice...). More training, both practical and theoretical, and more time to perform tasks were called for. CONCLUSIONS: The strategy to improve health professionals' ethical approach is to increase their Bioethics training. Very little bioethics training is provided for social work/health professionals by primary care management in the Community of Madrid. PMID- 15802112 TI - [Impact of common reasons for consultation on the feeling of vitality and the physical performance of the over-65s]. AB - OBJECTIVES: To measure two functional dimensions (vitality and physical function) involved in the quality of life of the over-65s and to find what relationship they maintain with the commonest reasons for consultation. DESIGN: Cross sectional, descriptive study. SETTING: Primary care. PARTICIPANTS: Randomised sample of 179 patients over 65 from 14 primary care clinics in Navarra. METHODS: SF-36 quality-of-life questionnaire and most common reasons for consultation. Personal and family details and ongoing drug prescription were also recorded. RESULTS: The most common reasons for consultation were insomnia (31.8%), arthrosis (48%), and urinary symptoms (16.2%). The greatest differences in the SF 36 scales occurred in patients with insomnia. In the multiple regression models, inverse associations were found for each of the reasons for consultation with the vitality and physical function dimensions. Vitality was associated with urinary symptoms, with an adjusted beta coefficient of -11.2 points (95% CI, -18.6 to 3.7). Insomnia was associated with significant decline in vitality and physical function, with beta of -7.7 points (95% CI, -13.9 to -1.5) and -10.3 points (95% CI, -19.1 to -1.6), respectively. Arthrosis symptoms behaved in a similar way. CONCLUSIONS: The most common pathologies or symptoms causing primary care consultations in the over-65s affect significantly the quality-of-life dimensions relating to the pursuit of normal daily life. PMID- 15802113 TI - [Indication of bone densitometry in women over 40 years of age]. AB - OBJECTIVE: To evaluate the indication of bone densitometry (BD) and to know the prevalence of risk factors (FR) for osteoporotic fracture in primary care. DESIGN: Cross-sectional, descriptive study. SETTING: Urban centre of health of Terrassa, Barcelona (52.000 h). PARTICIPANTS: Women over 40 years old than went to urgencies of the centre selected by consecutive sampling stratified by age (n=416). MAIN MEASUREMENTS: Registry of FR for osteoporotic fracture according to the criteria of the Agencia d'Avaluacio de Tecnologies Mediques (AATM), hygienic dietetic recommendations received and made BDs. RESULTS: The average age was of 60+/-13 years. The FR for osteoporotic fracture more prevalence were: low calcium intake (32.1%), increased age (27.5%), no lactation (24.9%), and precocious menopause (16.4%). They had received dietetic recommendations 35.7%, of exercise 65.5% and solar exhibition 18.1%. Of the 177 (42.7%) cases in that the BD was indicated only it made in 45 (25.4%) and of 237 (57.3%) without indication of BD they were made 41 (17.2%). To have made BD is related of statistically significant way to antecedent of osteoporotic fracture (P<.001) and to iatrogenic menopause (P<.001). The 41.2% of the BDs were asked for by the primary care physician, the 34.1% by the rheumatologist and the 16.5% by the gynaecologist. The indication of BD varies between the different scientific societies: AATM 42.7%, semFYC 45.7%, FORE 49.8%, SER 61.1%, and NOF 64.3%. CONCLUSIONS: Although FR of prevalence osteoporosis are most modifiable from primary care, a low degree of advice exists on healthful habits of life. A low accomplishment of BDs is detected and half is not adapted its indications according to the criteria of the AATM. There is great variability in the indication of BD between the different scientific societies. PMID- 15802114 TI - [Basic needs unsatisfied]. PMID- 15802115 TI - [Randomised clinical trial of an intensive intervention into life-styles of patients with hyperfibrinogenaemia in primary prevention of cardiovascular pathology in primary health care]. AB - OBJECTIVES: To study the effect of an intensive programme to modify life-style on levels of plasma fibrinogen in patients without cardiovascular pathology, with high fibrinogen and normal cholesterol levels. To analyse whether the effect on fibrinogen is independent, or otherwise, of the effect on lipids. DESIGN: Randomised clinical trial with a control. SETTING: 11 health districts in L'Hospitalet de Llobregat and Barcelona. PARTICIPANTS: 436 patients will be included, 218 individuals between 35 and 75 years old in each group, and without cardiovascular pathology (ischaemic cardiopathy, cerebral vascular accident or peripheral arteriopathy), with hyperfibrinogenaemia (fibrinogen > 300 mg/dL) and with plasma control < 250 mg/dL. INTERVENTIONS: One group of patients will receive an intensive intervention (in frequency and intensity of counselling and treatment) for life-style changes, i.e. stopping smoking, low-calorie diet in case of overweight or obesity, and physical exercise. The follow-up of the intervention group will be every 2 months. The control group will follow customary treatments. MEASUREMENTS: Levels of plasma fibrinogen. In addition, other relevant events will be recorded over a 2-year monitoring period: modification of risk factors, changes in quality of life, cardiovascular events or death. DISCUSSION: The introduction of an intensive primary prevention intervention (life-style changes) in patients with hyperfibrinogenaemia could be a more effective measure than the habitual intervention for reducing plasma fibrinogen figures. In addition, these measures could be translated into a reduction of cardiovascular risk and an improvement in the patient s quality of life. PMID- 15802116 TI - [Diabetes in pregnancy: screening, diagnosis, and monitoring at the health centre]. PMID- 15802117 TI - [The "false mobbing" syndrome: difficulties of diagnosis in primary care]. PMID- 15802118 TI - [Presence of Entamoeba histolytica in chronic urethritis]. PMID- 15802119 TI - [Do our patients approve of their acting general practitioner consolidating his/her job in the extraordinary OPE (Public Employment Offer) currently under way?]. PMID- 15802120 TI - [Adverse effect of beta-blockers or confusion]. PMID- 15802121 TI - [Aristotle and compulsory transfers]. PMID- 15802122 TI - [The examination of men's wits for the sciences: "Edipalaci" vs "ECOE" (Objective Clinical Examination)]. PMID- 15802123 TI - [Pregnancy and use of the safety belt]. PMID- 15802124 TI - Calculating the probability of detection for inhibitors in enzymatic or binding reactions in high-throughput screening. AB - In high-throughput screening (HTS) for drug candidates from a library containing tens of thousands to millions of chemical compounds, one problem is assessing the sensitivity of an assay for detecting compounds with a particular potency. For example, when looking for inhibitors of an enzyme, what is the potency of an inhibitor that will be readily detected by an enzyme inhibition assay? Similarly, when assessing compounds that inhibit binding between receptors and ligands or similar molecule-to-molecule interactions, what potency of an inhibitor will be readily detected? In this article, the well-established concepts of Michaelis Menten kinetics and Langmuir binding isotherms are combined with fundamental statistical principles to yield a measure of assay sensitivity. The approach is general and can be modified to accommodate situations where the reaction kinetics is known to be more complicated than situations described by the Michaelis-Menten and Langmuir equations. The calculations presented take into account the concentration of inhibitor used, the variability of the assay, the relationship between the K(m) or K(d) of the reaction and the substrate or ligand concentration used, the threshold or cutoff value used for determining "hits," and the number of replicates used in screening. PMID- 15802125 TI - Electroaddressed immobilization of recombinant HIV-1 P24 capsid protein onto screen-printed arrays for serological testing. AB - A serological chemiluminescent biochip was designed based on screen-printed electrode arrays composed of nine 1-mm(2) electrodes. Arrays were shown to be produced with good batch-to-batch reproducibility (standard deviations of 4.4 and 12.0% for ferricyanide oxidation potential and current, respectively) and very good reproducibility within a particular array (2.0 and 7.5% standard deviations for the same controls). Electrode arrays were used to electroaddress various bioconjugate structures comprising a recombinant HIV-1 P24 capsid protein (RH24K) in polypyrrole film. Entrapment of RH24K preimmobilized onto maleic anhydride-alt methyl vinyl ether copolymer was shown to be the more efficient immobilization procedure. This addressed sensing layer enabled the detection of anti-P24 antibodies at a concentration of 3.5 ng/ml through peroxidase-labeled anti-human immunoglobulin G reaction. The biochip was used to perform an HIV-1 serological test in human sera. HIV-1 seropositive and seronegative sera were easily discriminated using serum dilutions greater than 1/10,000. PMID- 15802126 TI - Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and investigation of the effect of dye concentration on amplification and DNA melting curve analysis. AB - Following the initial report of the use of SYBR Green I for real-time polymerase chain reaction (PCR) in 1997, little attention has been given to the development of alternative intercalating dyes for this application. This is surprising considering the reported limitations of SYBR Green I, which include limited dye stability, dye-dependent PCR inhibition, and selective detection of amplicons during DNA melting curve analysis of multiplex PCRs. We have tested an alternative to SYBR Green I and report the first detailed evaluation of the intercalating dye SYTO9. Our findings demonstrate that SYTO9 produces highly reproducible DNA melting curves over a broader range of dye concentrations than does SYBR Green I, is far less inhibitory to PCR than SYBR Green I, and does not appear to selectively detect particular amplicons. The low inhibition and high melting curve reproducibility of SYTO9 means that it can be readily incorporated into a conventional PCR at a broad range of concentrations, allowing closed tube analysis by DNA melting curve analysis. These features simplify the use of intercalating dyes in real-time PCR and the improved reproducibility of DNA melting curve analysis will make SYTO9 useful in a diagnostic context. PMID- 15802127 TI - Analysis of single nucleotide incorporation reactions by capillary electrophoresis. AB - Single nucleotide incorporation assays have been used to probe the kinetic parameters of many DNA and RNA polymerases. Traditionally, oligonucleotide primers are 5'-(32)P labeled using T4 kinase and annealed to a complementary template with a 5' overhang. To quantify the reaction kinetics, the products of the primer extension reactions are usually separated using denaturing polyacrylamide gel electrophoresis and quantified using a phosphorimager or other method to measure radioactivity. We have developed a method using a 5' fluorescently labeled oligonucleotide to examine the kinetics of single nucleotide incorporation catalyzed by recombinant human mitochondrial polymerase gamma (Pol gamma) holoenzyme. Using laser-induced fluorescence detection in the P/ACE MDQ instrument, primers 5' labeled with fluorescent probes such as 6 carboxyfluorescein can be rapidly separated and quantified. However, we also show that only select probes can be used, presumably due to unfavorable interactions between Pol gamma and certain 5' labels. PMID- 15802128 TI - Characterization of oxidized nicotinamide adenine dinucleotide (NAD(+)) analogues using a high-pressure-liquid-chromatography-based NAD(+)-glycohydrolase assay and comparison with fluorescence-based measurements. AB - A high-pressure-liquid-chromatography (HPLC)-based technique was developed to assess the oxidized nicotinamide adenine dinucleotide (NAD(+))-glycohydrolase activity of the catalytic domain of Pseudomonas exotoxin A containing a hexa-His tag. The assay employs reverse-phase chromatography to separate the substrate (NAD(+)) and products (adenosine 5'-diphosphate-ribose and nicotinamide) produced over the reaction time course, whereby the peak area of nicotinamide is correlated using a standard curve. This technique was used to determine whether the NAD(+) analogue, 2'-F-ribo-NAD(+), was a competing substrate or a competitive inhibitor for this toxin. This NAD(+) analogue was hydrolyzed at a rate of 0.2% that of NAD(+) yet retained the same binding affinity for the toxin as the parent compound. Finally, the rate that a fluorescent NAD(+) analogue, epsilon-NAD(+), is hydrolyzed by the toxin was also investigated. This analogue was hydrolyzed six times slower than NAD(+) as determined using HPLC. The rate of hydrolysis of epsilon-NAD(+) calculated using the fluorometric version of the assay shows a sixfold increase in reaction rate compared to that determined by HPLC. This HPLC based assay is adaptable to any affinity-tagged enzyme that possesses NAD(+) glycohydrolase activity and offers the advantage of directly measuring the enzyme catalyzed hydrolytic rate of NAD(+) and its analogues. PMID- 15802129 TI - The application of quantum dots as fluorescent label to glycoarray. AB - A sensitive, specific, and rapid method for the detection of carbohydrate-protein interactions was demonstrated using quantum dots (QDs) as a fluorescence label coupled with protein. 1,3-Dipolar cycloaddition between azide and alkyne was exploited to attach alpha-d-glucopyranoside to a C(14) hydrocarbon chain that noncovalently binds to the microtiter well surface, and the product formation was detected by both electrospray ionization-mass spectrometry (ESI-MS) and QD- (or fluorescein isothiocyanate (FITC))-conjugated lectin binding. It indicated that the peak intensity of the fluorescence emission was proportional to the initial concanavalin A (Con A) concentration in the range of 2 x 10(-3) micromol/L to 2 x 10(-2)mmol/L with a detection limit at least 100 times lower than that of the FITC-based method. PMID- 15802130 TI - Equilibrium studies of a fluorescent tacrolimus binding to surfactant protein A. AB - Tacrolimus (FK506) is a hydrophobic immunosuppressive agent used in kidney, liver, and lung transplantation. The objective of this study was to characterize the binding of FK506 to surfactant protein A (SP-A), an abundant lipoprotein found in the alveolar fluid that functions as part of the innate immune system in the lung. We have synthesized a novel derivative of FK506 in which a dansyl moiety was covalently bound via cadaverine to the C22 position of the FK506 molecule (DNS-FK). Using the fluorescence and anisotropy properties of DNS-FK, we demonstrated that tacrolimus avidly binds to SP-A with an apparent equilibrium association constant (K(app)) of 10(7)M(-1) and a Gibbs binding free energy of 40 kJ mol(-1)K(-1). Derivatization of FK506 at the C22 position did not block FK506 binding to the cytosolic immunophilin FK506-binding protein (FK-BP) or human serum albumin (HSA), both used as controls of tacrolimus-binding proteins. K(app) for FK-BP/DNS-FK and HSA/DNS-FK complexes were 1.5 x 10(7) and 10(7)M(-1), respectively. The high sensitivity of this analytical technique makes it suitable for binding analysis of FK506 to proteins. PMID- 15802131 TI - Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide sensitive europium probe. AB - A method for optical imaging of the activity of glucose oxidase (GOx) using a fluorescent europium(III) tetracycline probe for hydrogen peroxide is presented. A decay time in the microsecond range and the large Stokes shift of 210 nm of the probe facilitate intensity-based, time-resolved, and decay-time-based imaging of glucose oxidase. Four methods for imaging the activity of GOx were compared, and rapid lifetime determination imaging was found to be the best in giving a linear range from 0.32 to 2.7 m Unit/mL. The detection limit is 0.32 m Unit/mL (1.7 ng mL(-1)) which is similar to that of the time-resolved (gated) imaging using a microtiterplate reader. Fluorescent imaging of the activity of GOx is considered to be a useful tool for GOx-based immunoassays with potential for high-throughput screening, immobilization studies, and biosensor array technologies. PMID- 15802132 TI - Using surface plasmon resonance to directly determine binding affinities of combinatorially selected cyclopeptides and their linear analogs to a streptavidin chip. AB - In our recent report, several HPQ-containing streptavidin ligands were identified from a structurally constrained combinatorial library, and the relative affinities in IC(50) of these tight-binding ligands were revealed by a captured enzyme-linked immunosorbent assay. In the present work, surface plasmon resonance was employed to directly evaluate the binding affinities between immobilized streptavidin and combinatorially selected ligands. The equilibrium dissociation constants and kinetic on/off rates of a previously identified N-to-side chain and newly synthesized N-to-C cyclopeptides were readily deduced using Scatchard analysis and computational simulation. It was found that both cyclopeptides bound streptavidin far more tightly than its linear counterpart ( approximately 1000 fold), while the reversed (QPH) linear and cyclic peptidyl ligands were hardly recognized by streptavidin. Consequently, not only was the binding specificity of synthetic ligands distinguished qualitatively but also the entropic advantage of conformationally constrained cyclopeptides over their linear forms was demonstrated quantitatively by surface plasmon resonance. PMID- 15802133 TI - A fluorogenic substrate for the continuous assaying of aryl sulfatases. AB - The most common fluorogenic substrate for assaying aryl sulfatases (ARSs) is 4 methylumbelliferyl sulfate (MUS). However, ARSs operate optimally at pH values that are less than the pK(a) (7.8) of the reaction product of MUS, 4 methylumbelliferone (4-MU). Thus, a major disadvantage of this assay is that it is usually run in a discontinuous mode due to the need for basification of the reaction mixture to achieve complete ionization of the phenolic products and maximum fluorescence. To circumvent this problem, 6,8-difluoro-4 methylumbelliferyl sulfate (DiFMUS) was prepared and examined as a substrate for ARSs. The product of the reaction is 6,8-difluoro-4-methylumbelliferone, a known coumarin with fluorescent properties equal to those of 4-MU, and has a pK(a) of 4.9. This allowed for the continuous assaying of human placental ARSs A, B, and C, which operate optimally between pH 5.0 and pH 7.0. Furthermore, DiFMUS exhibited a lower K(m) (up to 20-fold) for the ARSs than did MUS; for ARSA and ARSB, it exhibited a greater V(max) than did MUS. This substrate should have considerable utility for the continuous assay of ARS activity. PMID- 15802134 TI - Probing conformational changes of proteins by quantitative second-derivative infrared spectroscopy. AB - Probing protein conformational changes plays a crucial role in protein structure and function studies. However, the lack of efficient biophysical techniques makes it difficult to obtain the distinct behaviors of different secondary structure elements in a protein upon perturbation. This paper presents a discussion of the two major problems, the effect of sidelobes and different half-width at half height (HWHH) values, encountered in quantitative second-derivative infrared (QSD IR) spectroscopy and introduces the development of two criteria for checking the validity of the results obtained using the QSD-IR method. It was found that neither the sidelobes nor the HWHH significantly affected the quantitative result of protein conformational changes by using poly-l-lysine and hemoglobin as model proteins. A case study of bovine serum albumin (BSA) thermal aggregation suggested that the thermal transition of BSA was a process involving sequential events, and the two helical components were found to have a distinct response to heat perturbation. These results were confirmed by two-dimensional infrared correlation spectroscopy and by results in literature, suggesting that the QSD-IR method might be a potentially powerful tool to probe the distinct response of different secondary structures to perturbation. PMID- 15802135 TI - Comparison of metabolic profiles from serum from hepatotoxin-treated rats by nuclear-magnetic-resonance-spectroscopy-based metabonomic analysis. AB - Hepatotoxicities were induced in rats using alpha-naphthylisothiocyanate (ANIT), carbon tetrachloride (CCl(4)), and hydrazine (HYD). Male Wistar rats were treated with three typical hepatotoxins, and serum samples were collected after 48 h. Biochemical effects of these toxins on plasma composition were evaluated by high resolution (1)H nuclear magnetic resonance (NMR) spectroscopy of serum. The biochemical effects of CCl(4) were characterized by an elevated level of 3-d hydroxybutyrate (HB), acetoacetate (Aca), and creatinine (Cn) in serum, and ANIT led to increases in the amounts of low-density lipoprotein (LDL), alanine, acetate, glycoprotein, succinate, Cn, acetone, 3-d-hydroxybutyrate, and Aca. For the HYD-treated group, LDL, HB, acetate, and Cn were obviously increased in serum. The region delta 0.0-10.0 of each spectrum was segmented into 0.04 ppm. The area under the spectrum was calculated for each segmented region and expressed as an integral value. After removal of the water signal (delta 4.6-5.0) the remaining 235 intensity-related descriptors were used for the pattern recognition analysis. Principal component analysis was used to visualize the similarities and differentiations in biochemical profiles of serum from the rats treated with various hepatotoxins. This work showed the power of the combination of NMR and pattern recognition for the study of biochemical effects of xenobiotics. PMID- 15802136 TI - Development of a fast kinetic method for the determination of carboxypeptidase U (TAFIa) using C-terminal arginine containing peptides as substrate. AB - Carboxypeptidase U (CPU, TAFIa) is a novel determinant of the fibrinolytic rate. It circulates in blood as an inactive zymogen, procarboxypeptidase U, which is activated during the process of coagulation and fibrinolysis. CPU has a very short half-life at 37 degrees C. Its intrinsic instability complicates the determination of kinetic parameters of different substrates using an endpoint method. We developed a fast kinetic assay for measuring continuously the release of the C-terminal arginine by CPU independent of the nature of the substrate peptide used, allowing us to perform substrate specificity studies of CPU. This method uses arginine kinase, pyruvate kinase, and lactate dehydrogenase as auxiliary enzymes. The CPU activities measured using this kinetic assay were in the range of 97-103% of those determined with our HPLC-assisted reference assay, and the obtained K(m) and k(cat) values for hippuryl-l-arginine and bradykinin were in good accordance with those described in the literature. As expected, no arginine cleaving was seen using dipeptides and peptide substrates with a proline in the penultimate position. The presented kinetic assay enables the fast screening of substrates with a C-terminal arginine and is a valuable new tool for the kinetic evaluation of both synthetic and physiological substrates of CPU. PMID- 15802137 TI - Simultaneous quantitative analysis of ceramide and sphingosine in mouse blood by naphthalene-2,3-dicarboxyaldehyde derivatization after hydrolysis with ceramidase. AB - Ceramide and sphingosine are sphingolipids with important functional and structural roles in cells. In this paper we report a new enzyme-based method to simultaneously quantify the levels of ceramide and sphingosine in biological samples. This method utilizes purified human recombinant acid ceramidase to completely hydrolyze ceramide to sphingosine, followed by derivatization of the latter with naphthalene-2,3-dialdehyde (NDA) and quantification by reverse-phase high-performance liquid chromatography. The limits of detection for sphingosine NDA and ceramidase-derived sphingosine-NDA were 9.6 and 12.3 fmol, respectively, and the limits of quantification were 34.2 and 45.7 fmol, respectively. The recovery of sphingosine and ceramide standards quantified by this assay were between 95.6 and 104.6%. The relative standard deviations for the intra- and interday sphingosine assay were 2.1 and 4.5%, respectively, and those for the ceramide assay were 3.3 and 4.1%, respectively. To validate this procedure, we quantified ceramide and sphingosine in mouse plasma, white blood cells, and hemoglobin, the first reported time that the amounts of these lipids have been documented in individual blood components. We also used this technique to evaluate the ability of a novel ceramide analog, AD2646, to inhibit the hydrolytic activity of acid ceramidase. The results demonstrate that this new procedure can provide sensitive, reproducible, and simultaneous ceramide and sphingosine quantification. The technique also may be used for determining the activity and inhibition of ceramidases and may be adapted for quantifying sphingomyelin and sphingosine-1-phosphate levels. In the future it could be an important tool for investigators studying the role of ceramide/sphingosine metabolism in signal transduction, cell growth and differentiation, and cancer pathogenesis and treatment. PMID- 15802139 TI - Development of an oral biosensor for salivary amylase using a monodispersed silver for signal amplification. AB - An amperometric biosensor for monitoring the level of protein amylase in human saliva is described. A novel design and the preparation of amylase antibodies and antigens, essential for the development of the biosensor, are reported. The biosensor sensing elements comprise a layer of salivary antibody (or antigen) self-assembled onto Au-electrode via covalent attachment. Molecular recognition between the immobilized antibody and the salivary amylase proteins was monitored via an electroactive indicator (e.g., K(3)Fe(CN)(6)) or a monodispersed silver layer present in solution or electrochemically deposited onto the solid electrode. This electroactive indicator was oxidized or reduced and the resulting current change provided the analytical information about the concentration of the salivary proteins. The limit of detection of 1.57 pg/ml was obtained, in comparison to detection limits of 4.95 pg/ml obtained using potassium ferrocyanide as the redox probe and 10 ng/ml obtained using enzyme-linked immunosorbent assay. Cross-reactivity was tested against cystatin antibodies and was found to be less than 2.26%. PMID- 15802138 TI - Fabrication of DNA microarrays onto poly(methyl methacrylate) with ultraviolet patterning and microfluidics for the detection of low-abundant point mutations. AB - We have developed a simple ultraviolet (UV)-photomodification protocol using poly(methyl methacrylate) and polycarbonate to produce functional scaffolds consisting of carboxylic groups that allow covalent attachment of amine terminated oligonucleotide probes to these surface groups through carbodiimide coupling. Use of the photomodification procedure coupled to microfluidics allowed for the rapid generation of medium-density DNA microarrays. The method reported herein involves the use of poly(dimethylsiloxane) microchannels reversibly sealed to photomodified poly(methyl methacrylate) surfaces to serve as stencils for patterning the oligonucleotide probes. After array construction, the poly(dimethylsiloxane) stencil is rotated 90 degrees to allow interrogation of the array using microfluidics. The photomodification process for array fabrication involves only three steps: (1) broadband UV exposure of the polymer surface, (2) carbodiimide coupling of amine-terminated oligonucleotide probes to the surface (via an amide bond), and (3) washing of the surface. The density of probes attached to this activated surface was found to be approximately 41pmolcm( 2), near the steric-saturation limit for short oligonucleotide probes. We demonstrate the use of this procedure for screening multiple KRAS2 mutations possessing high diagnostic value for colorectal cancers. A ligase detection reaction/universal array assay was carried out using parallel detection of two different low-abundant DNA point mutations in KRAS2 oncogenes with the allelic composition evaluated at one locus. Four zip code probes immobilized onto the poly(methyl methacrylate) surface directed allele-specific ligation products containing mutations in the KRAS2 gene (12.2D, 12.2A, 12.2V, and 13.4D) to the appropriate address of a universal array with minimal amounts of cross hybridization or misligation. PMID- 15802140 TI - Rate-of-dialysis technique: theoretical and practical aspects. AB - In the rate-of-dialysis technique, chemical equilibrium exists throughout the experiment between the contents of the two halves of a dialysis cell. Initially, however, a trace amount of labeled dialysand is present in one of the half-cells, and the dialysis of that is followed as a function of time. The method is able to reveal whether a compound is stable under the conditions used or whether it is affected, for example, by aggregation, precipitation, oxidation, or hydrolysis. The rate constants for six inorganic ions and 22 organic molecules were determined, and no simple relations were found between rate constants and functions of molar masses. However, in two homologous series of compounds, a linear relationship between rate constant and molar mass was found, allowing for a preliminary identification of an unknown substance. Whether the presence of a macromolecule results in binding of the dialysand is easily tested for because in that case its rate constant is sharply diminished. High-affinity binding was studied using octanoate and laurate as ligands and human serum albumin as binding protein. The primary binding constants determined were similar to those obtained by equilibrium dialysis. However, the rate-of-dialysis technique is much faster, and its results are less scattered. The technique is well suited for binding studies performed with high protein concentrations because it is not affected by any osmosis or Donnan effects. PMID- 15802141 TI - Forster resonance energy transfer in liposomes: measurements of transmembrane helix dimerization in the native bilayer environment. AB - The lipid bilayer vesicle is a model of the cellular membrane. Even in this simple system, however, measuring the thermodynamics of membrane protein association is a challenge. Here we discuss Forster resonance energy transfer (FRET) in liposomes as a method to probe the dimerization of transmembrane helices in a membrane environment. Although the measurements are labor intensive, FRET in liposomes can be measured accurately provided that attention is paid to sample homogeneity and sample equilibration. One must also take into account statistical expectations and the FRET that results from random colocalization of donors and acceptors in the bilayer. Without careful attention to these details, misleading results are easy to obtain in membrane FRET experiments. The results that we obtain in model systems are reproducible and depend solely on the concentration of the protein in the bilayer (i.e., on the protein-to-lipid ratio), thereby yielding thermodynamic parameters that are directly relevant to processes in biological membranes. PMID- 15802142 TI - Fluorometric TLC assay for evaluation of protein kinase inhibitors. AB - A fluorometric assay for measuring protein kinase activity has been developed. The assay is based on the separation of fluorescently marked substrate 5 carboxytetramethylrhodamine-kemptide (5-TAMRA-kemptide) from its phosphorylated counterpart by TLC and quantification of the product ratiometrically by fluorescence imaging. The utility of the assay was demonstrated by measuring the activity of cAMP-dependent protein kinase. 5-TAMRA-kemptide was characterized as a substrate of this kinase by the kinetic parameters K(m)(app) and V(max). The attachment of 5-TAMRA dye to the N terminal of kemptide decreased the K(m)(app) value but did not have a significant effect on the rate and stoichiometry of the phosphorylation reaction. The inhibitory potency of three known inhibitors was evaluated with the new assay. The closeness of the obtained inhibitory activities of the compounds to the activities determined with the phosphocellulose paper binding assay, as well as the Z' factor value of 0.5, demonstrates the reliability of the new assay for evaluation of inhibitors of protein kinases. PMID- 15802143 TI - In vivo quantification of intracellular amino acids and intermediates of the methionine pathway in Corynebacterium glutamicum. PMID- 15802144 TI - Gradient polymerase chain reaction performance using regular thermal cycle machine. PMID- 15802145 TI - Restriction analysis of recombinant plasmids from colonies in less than 30 min. PMID- 15802146 TI - Measurement of glucose concentrations in human plasma using a glucose biosensor. PMID- 15802147 TI - Molecular transplantation: delivery of membranous proteins onto live cells. PMID- 15802148 TI - The politics of childbirth in the context of conflict: policies or de facto practices? AB - The impact of warlike conditions on the lives of Palestinian women and children is far reaching. Beginning in September 2000, curfews, closures, siege and the parceling up of the country into small isolated entities have all led to a lack of access to basic maternity services, rendering pregnant women and their newborns a highly vulnerable group. Because any discussion of childbirth in the Occupied Palestinian Territories (OPT) cannot be separated from the larger historical context of international health politics, we begin with a brief review of international historical trends in childbirth policies, focusing on the relationship between discourse in the developed and developing world contexts to show how these models have intersected and diverged. We point to the similarities between the OPT and other developing countries, but also highlight the specificities that characterize the Palestinian experience today that include local political systems, medical dominance, professional group interests and the politics of gender, as well as the legacy of colonialism intertwined with an ongoing national conflict. We then provide a review of the history of childbirth in the OPT and analyze the various forces that led to the emergence of today's chaotic and contradictory de facto policies and practices. By assessing the health policy environment, we demonstrate the seeming impossibility of developing national level childbirth policies, given the current political conditions and a mix of other determinants that are not all within Palestinian control. Finally, we emphasize the importance of establishing a process as opposed to a blueprint of health policy-making based on people's immediate and long-term needs in all areas of the country. We also propose interim measures that rest on the notion of developing decentralized sub-strategies relevant to different zones of political reality and stages of system and human resource development, aiming at combining survival imperatives with those of improving women's birth experiences and women's health. PMID- 15802149 TI - Cancer treatment at home or in the hospital: what are the costs for French public health insurance? Findings of a comprehensive-cancer centre. AB - OBJECTIVE: The objective of this study was to evaluate the cost of home-cancer healthcare programs and their potential interest for public health insurance as compared to inpatient cancer care. PATIENTS AND METHODS: The study was conducted at the Centre Leon Berard (CLB), a comprehensive cancer centre in Lyon, France. Hospitals at home patients were monitored by nurses and oncologists from the CLB. All patients, who received home treatment over a 15-day period in 2001, were included in the study. Patients were broken down into groups according to the type of healthcare required and the corresponding impact on health insurance expenditure. For each of these patients, a fictive-hospital stay was then reconstructed, which corresponded to the inpatient hospital care that would have been required during the observation period, had hospital at home not been available. RESULTS: The average cost of hospital at home was significantly lower than the corresponding estimated cost for treatment at the hospital (776.6 versus 2012.5, P < 0.001). This difference was particularly high for patients in the "palliative care" group (N = 33) (1201.7 versus 3489.7, P < 0.001), whereas in the "chemotherapy" group, results were not significantly different (N = 34) (225.5 versus 318). CONCLUSION: Our study suggests that hospitalisation alternatives can generate substantial savings for public health insurance in France. PMID- 15802150 TI - Inequality in willingness-to-pay for community-based health insurance. AB - The purpose was to provide information for devising community-based health insurance (CBI) policies that reduce inequality in enrolment and further inequality in access to health services. A two-stage cluster sampling was used in the household survey. Inequalities in willingness-to-pay (WTP) for CBI are examined by expenditure quintile using data collected from a household survey. Interviews were conducted with 2414 individuals, 705 of whom were household heads. A bidding game method was used to elicit WTP. Individuals and households were assigned to 6-month expenditure quintiles. We found that mean and median individual WTP for CBI was significantly higher for higher spending quintiles, as was mean and median household WTP. The curves of cumulative percentage of individual and household WTP shifted rightwards for higher quintiles, implying that at any given premium the lower the quintile the lower the uptake of CBI. The Gini coefficient for individual WTP and household WTP was 0.15 and 0.08, respectively, and for individual 6-month expenditure and household 6-month expenditure is 0.68 and 0.63, respectively. The results imply that the premium needs to be adjusted for income; otherwise, a lower proportion of poor people will enrol in CBI and without exemptions or subsidies the poor will have less access to health services than the rich. Thus, exemptions and subsidies for the poor for enrolling in CBI are an important issue for decision-makers with an objective of improving equity of health and helping the poor to break out of the cycle of poverty. Since the distribution of WTP by household is less unequal than the distribution of WTP by individuals, the household might be a better unit of enrolment in terms of equity than the individual. PMID- 15802151 TI - Improvement in maternal health literacy among pregnant women who did not complete compulsory education: policy implications for community care services. AB - This paper examined factors that influence the improvement in maternal health literacy among pregnant women in Paraguay, including those who did not complete compulsory education but participated in a community-based antenatal care program. Structured interviews were conducted to evaluate the pregnant women's maternal health literacy during their first, second, and third visits to the program in the Caazapa Region. The associations between individual maternal health knowledge scores and its gains, healthcare personnel capabilities, available health facility equipment, community social network, and living environment were analyzed by multiple regression analysis. The mean maternal health knowledge score from 124 women who completed three-consecutive assessments increased between the first and third interviews. Higher capabilities of healthcare personnel and better living environment were significantly related to gains in the maternal health knowledge score (p<0.01). Wider application of a community-based antenatal care program to meet the needs of those who are functionally illiterate in the standard language of the country, training for community healthcare personnel to improve capabilities, and resources for social network in the community would contribute to the improvement in maternal health literacy. PMID- 15802152 TI - The integration of medical specialists in hospitals. Dutch hospitals and medical specialists on the road to joint regulation. AB - Medical specialists have--notwithstanding governmental policy--acquired a firmly established position in policy-making in Dutch hospitals. In all general hospitals the Executive Board and the Staff Executive frequently and systematically negotiate their interests. Three conclusions may be drawn on the basis of the research reported on in this article. In the first place, it is the primary objective of the medical staff executive to protect the interests of the medical domain against managerial dominance. In the second place, developments suggest that the parties are slowly but surely switching to the joint regulation of overlapping interests. Finally, the trend towards joint regulation is an enormous challenge for medical staff governance. PMID- 15802153 TI - Living donor liver transplantation: the historical and cultural basis of policy decisions and ongoing ethical questions. AB - Adult-to-adult living donor liver transplantation (LDLT) is in a state of flux. Technical innovations and demand have outpaced internal and external regulatory efforts. This has led to a wide array of centers performing LDLT for a variety of indications without clear evidence on the risks to the donor or recipient or the system as a whole. The birth from necessity of LDLT in Asia has led to the extrapolation of the technique in America and Europe that has not been sufficiently studied in the appropriate populations. While there is a clear benefit in some patients, the appropriate donors and recipients have not been defined. Regulatory and ethical consideration should be focused on minimizing acceptable risk in donors and recipients and expanding the investigation into the costs and outcomes of this challenging procedure. The recently funded adult-to adult living donor liver transplantation cohort sponsored by the National Institutes of Health aims to answer some of these questions over the next five years. PMID- 15802154 TI - Equity in utilization of cancer inpatient services by income classes. AB - Equity in health care services has been prioritized on the Korean government's policy agenda since the government-driven national health insurance achieved universal coverage in 1989 along with the final inclusion of the self-employed as beneficiaries. The purpose of this study is to identify disparities in the utilization of health care services, especially cancer inpatient services among different income groups in Jeju Island of South Korea. We analyzed the national health insurance data about qualification of beneficiaries and utilization of health care services consumed by Jeju Island's residents for 1 year of period (from January to December 2000) and acquired their utilization features of cancer inpatient services. The independent variable was 10 different income levels according to the national health insurance fee imposed on each household in 2000. The dependent variable was the volume of cancer inpatient services utilized, that was measured by admission days and costs for treatment. The utilization of cancer inpatient services in the 10 different income groups was analyzed in three geographical categories of medical institutions: (1) within Jeju Island; (2) outside Jeju Island; (3) South Korea in total. We calculated the concentration indices of cancer inpatient services utilization in admission days and cost as a pair amongst these three geographical categories each. Both of the concentration indices were negative for the category of 'within Jeju Island', positive for that of 'outside Jeju Island', and positive for that of 'South Korea in total'. These results suggest the relatively poor experience considerable inequality in the utilization of cancer inpatient services in Jeju Island, because lower income groups have higher incidence rates in most cancers and inevitably have more needs in health services. PMID- 15802155 TI - Tackling excessive waiting times for elective surgery: a comparative analysis of policies in 12 OECD countries. AB - This paper compares policies to tackle excessive waiting times for elective surgery in 12 OECD countries. It is found that waiting times may be reduced by acting on the supply of or on the demand for surgery (or both). On the supply side, evidence suggests that both capacity and financial incentives towards productivity can play an important role. On the demand side, inducing a raising of clinical thresholds may reduce waiting times but may also provoke tension between clinicians and policy makers. Preliminary evidence also suggests that an increase in private health insurance coverage may reduce waiting times. PMID- 15802156 TI - Researching income and income distribution as determinants of health in Canada: gaps between theoretical knowledge, research practice, and policy implementation. AB - The research identified gaps in Canadian knowledge and research activity concerning the roles that income and its distribution play in Canadians' population health. 241 Canadian research studies on income and health were considered along eight taxonomies: conceptualization of income or its proxies; theoretical underpinnings; income distribution measures; health measures; who/what was studied, pathways mediating between income and health; complexity of these pathways; research design; and presence of policy implications. The study identified the following areas of weakness: (a) poor conceptualization of income and the means by which it influences health; (b) lack of longitudinal studies of the impact of income-related issues upon health across the life-span; (c) lack of linked data bases that allow complex analyses of how income and related issues contribute to health and well-being, and (d) little inter-disciplinary work in identifying pathways mediating the income and health relationship. Advances in health policy to address the health effects of income and its distribution requires a research infrastructure that draws upon recent theoretical developments in the area and is able to access data sources to test these advanced conceptualizations. PMID- 15802157 TI - Primary care teams: New Zealand's experience with community-governed non-profit primary care. AB - Community-governed non-profit primary care organisations started developing in New Zealand in the late 1980s with the aim to reduce financial, cultural and geographical barriers to access. New Zealand's new primary health care strategy aims to co-ordinate primary care and public health strategies with the overall objective of improving population health and reducing health inequalities. The purpose of this study is to carry out a detailed examination of the composition and characteristics of primary care teams in community-governed non-profit practices and compare them with more traditional primary care organisations, with the aim of drawing conclusions about the capacity of the different structures to carry out population-based primary care. The study used data from a representative national cross-sectional survey of general practitioners in New Zealand (2001/2002). Primary care teams were largest and most heterogeneous in community-governed non-profit practices, which employed about 3% of the county's general practitioners. Next most heterogeneous in terms of their primary care teams were practices that belonged to an Independent Practitioner Association, which employed the majority of the country's general practitioners (71.7%). Even though in absolute and relative terms the community-governed non-profit primary care sector is small, by providing a much needed element of professional and organisational pluralism and by experimenting with more diverse staffing arrangements, it is likely to continue to have an influence on primary care policy development in New Zealand. PMID- 15802158 TI - Do online pharmacies fit European internal markets? AB - The aim of this article is to consider the suitability of online pharmacies into European internal market area. This required considering the models of present online pharmacies in respect to the existing legislation. Data on online pharmacy settings was collected by looking some online pharmacies, which were found by using Goggle search machine with term "online pharmacy" and by studying websites of some well-known online pharmacies. European legislation and policy were studied from European Union's official website. Online drug markets seem to be increasing in popularity for reasons related to their ready availability and cost benefits. Few online pharmacies are based in Europe, yet online markets are worldwide. Community legislation does not stipulate on the legality of online pharmacies on European internal markets. Instead Community legislation offers framework for electronic commerce that could also include online pharmacy practise. National legislation, however, may rule them out either directly or indirectly. Regardless of European internal markets online pharmacies' cross border operations are particularly complicated. Preliminary ruling from the European Court of Justice concerning one European online pharmacy's cross-border practise is awaited 2003-2004 and will offer some aspects for future. PMID- 15802160 TI - The systemic response to lung infection. AB - The host response to microbial invasion in the lung must be sufficiently vigorous to allow for microbial eradication but appropriately controlled to prevent spillover of the response into the systemic circulation. Although inflammatory responses in pneumonia are generally compartmentalized, microbial and host factors can promote disordered systemic responses to lung infection. Assessment of the magnitude of the systemic inflammatory response in pneumonia is of limited clinical value, and attempts to suppress this response have failed to improve clinical outcomes. The systemic inflammatory response that occurs in sepsis and other critical illnesses can substantially impair lung innate and acquired immunity. Mechanisms of critical illness-induced immunoparalysis have been incompletely characterized and are the focus of ongoing clinical and basic investigations. PMID- 15802161 TI - Pulmonary host defenses and factors predisposing to lung infection. AB - Hospital-acquired pneumonia remains a leading cause of nosocomial infections and is associated with significant rates of morbidity and mortality. Multiple host-, pathogen-, and intervention-related factors contribute to the development of pneumonia in this setting. Potentially deleterious impacts of illness or therapeutic modalities impair the host's ability to resist the development of pneumonia while in the hospital and predispose to a spectrum of pathogens of greater virulence than found in the community. Recognition of factors contributing to pneumonia in the hospital setting may offer the opportunity to intervene, minimizing, to the extent possible, the serious consequences of this nosocomial complication. PMID- 15802163 TI - Genetic susceptibility to pneumonia. AB - The persistent mortality from community-acquired pneumonia may be explained by genetic predisposition. Specific mutations or polymorphisms in host response genes that are associated with adverse outcomes from infection can be grouped into four categories: antigen recognition, proinflammatory responses, anti inflammatory responses, and effector mechanisms. Mannose-binding lectin polymorphisms have a more dominant role in pneumonia when compared with other pattern recognition molecules such as the toll-like receptors. The roles of TNF and lymphotoxin alpha polymorphisms remain unclear despite extensive study. IL-10 and IL-1 receptor antagonist polymorphisms have an important role in the anti inflammatory response. Specific organ dysfunction, such as ARDS or DIC, may be related to polymorphisms in specific effector genes. PMID- 15802162 TI - Community-acquired pneumonia and sepsis. AB - Sepsis is a frequent and often fatal complication of pneumonia. This article discusses the epidemiology, pathophysiology, and treatment of sepsis in the setting of pneumonia. Particular consideration is given to the role of mechanical ventilation in amplifying organ dysfunction in sepsis and to treatments that have positive effects on sepsis mortality and respiratory dysfunction. PMID- 15802164 TI - Pathophysiology of pneumonia. AB - The development of pneumonia requires that a pathogen reach the alveoli and that the host defenses are overwhelmed by microorganism virulence or by the inoculum size. The endogenous sources of microorganisms are nasal carriers, sinusitis, oropharynx, gastric, or tracheal colonization, and hematogenous spread. Other external sources of contamination, such as intensive care unit workers, aerosols, or fibrobronchoscopy, must be considered as accidental. PMID- 15802165 TI - Etiology of community-acquired pneumonia. AB - Community-acquired pneumonia (CAP) is a serious lower respiratory tract infection associated with significant morbidity and mortality that is characterized by disputes over diagnostic evaluations and therapeutic decisions. With the widespread use of broad-spectrum antimicrobial agents and the increasing number of immunocompromised hosts, the etiology and the drug resistance patterns of pathogens responsible for CAP have changed. Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis remain the leading causes of CAP in immunocompetent patients. Opportunistic infections with organisms such as Pneumocystis jiroveci and Mycobacterium tuberculosis and other opportunistic fungal pneumonias should also be considered in the differential diagnosis of CAP in immunocompromised patients. This article examines the current peer-reviewed literature on etiology, risk factors, and outcomes of patients with CAP. PMID- 15802166 TI - Antimicrobial resistance and treatment of community-acquired pneumonia. AB - This article discusses the problem of antimicrobial resistance and how it affects the management of community-acquired pneumonia (CAP). The discussion is limited to infection with Streptococcus pneumoniae and to the treatment of patients hospitalized in a medical ward or an intensive care unit because of pneumococcal CAP. PMID- 15802167 TI - Antimicrobial treatment of community-acquired pneumonia. AB - Community-acquired pneumonia (CAP) is a clinical diagnosis that has a significant impact on health care management around the world. Early clinical suspicion and prompt empiric antimicrobial therapies are mandatory in patients with CAP. This article provides a review of recent studies and guidelines addressing antimicrobial therapy for hospitalized patients with CAP. PMID- 15802168 TI - Mechanisms of resistance of bacteria causing ventilator-associated pneumonia. AB - The common causes of ventilator-associated pneumonia (Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter cloacae, Acinetobacter spp) are frequently resistant to multiple antibiotic classes. S aureus develops resistance to all beta-lactam antibiotics by producing a modified penicillin-binding protein. Linezolid resistance arises by way of mutations in the 23S ribosomal subunit. Antibiotic resistance in gram-negative bacilli usually arises by way of beta-lactamase production, upregulation of efflux pumps, or loss of outer membrane proteins. The ability of bacteria to develop and disseminate new mechanisms of antibiotic resistance may outstrip the availability of new antibiotic classes. PMID- 15802169 TI - Microbiologic diagnosis of ventilator-associated pneumonia. AB - No single test will provide all the information needed to diagnose ventilator associated pneumonia (VAP), and the clinician must use multiple clinical criteria for the diagnosis. The use of culture techniques allows one to enhance the sensitivity and specificity of the diagnosis of pneumonia. In addition, the clinician can be kept up to date about potential new pathogens as they occur in the intensive care unit. Knowledge of the local flora allows one to make a better choice of empiric therapy for VAP. PMID- 15802170 TI - Etiology of ventilator-associated pneumonia. AB - This review focuses on the top ten causes of ventilator-associated pneumonia (VAP), updating an earlier study. These pathogens have specific risk factors, different patterns of clinical resolution, and a wide range of attributable mortality. The discussion herein analyzes these aspects, placing particular emphasis on risk factors, attributable mortality, resistance, and the implications for management. PMID- 15802171 TI - Antimicrobial treatment of hospital-acquired pneumonia. AB - This article highlights the importance of providing adequate empirical antibiotic therapy for hospital-acquired pneumonia and avoiding the excessive use of antibiotics. To meet these goals, a strategy for the management of suspected ventilator-associated pneumonia should include obtaining reliable pulmonary specimens for direct microscope examination and cultures before new antibiotics are administered. PMID- 15802172 TI - Acute lung injury and bacterial infection. AB - The relationships between acute lung injury and bacterial infection are complex. Indeed, sepsis and in particular pneumonia are leading causes of acute lung injury. Bacterial superinfection of the lung is a frequent complication of acute lung injury. Because of impaired host defenses and prolonged mechanical ventilation, more than one third of patients with the acute respiratory distress syndrome acquire ventilator-associated pneumonia, with resistant pathogens in most instances. This complication is responsible for more than a doubling of the time on mechanical ventilation but does not seem to increase mortality. PMID- 15802173 TI - Pneumonia in the solid organ transplant patient. AB - As the field of solid organ transplantation has grown, so has the importance of infectious complications in this select group of patients. Chronic immunosuppression compromises the natural host defenses that typically prevent lower respiratory tract infections and makes the solid organ transplant recipient especially susceptible to pneumonia. Evaluation of pneumonia in this population differs owing to the potential for opportunistic infections. Lung transplant recipients are particularly susceptible to pneumonia and pose unique diagnostic dilemmas. An understanding of the time line for the different key pathogens after transplantation aids the initial evaluation and management. PMID- 15802174 TI - Preventing pneumonia: the role for pneumococcal and influenza vaccines. AB - Streptococcus pneumoniae and influenza cause a wide spectrum of illness and result in substantial morbidity and mortality. They are significant public health concerns, and vaccines against both organisms exist. The pneumococcal polysaccharide vaccine has been available in its current form for over 20 years. Nonetheless, definitive evidence from prospective trials of its efficacy is lacking. Experts recommend that patients at high risk for pneumococcal infection and complications from this process be vaccinated. The role for revaccination remains controversial. Traditional influenza vaccine is composed of three virus types and decreases rates of serologically confirmed cases of influenza, hospital admissions for respiratory infection, and mortality. The pneumococcal and influenza vaccines are considered cost-effective options for pneumonia prevention. PMID- 15802175 TI - Prevention of pneumonia in the hospital setting. AB - Although the optimal approach to reducing ventilator-associated pneumonia (VAP) is unclear, recent studies indicate that mandatory education of health care workers caring for mechanically ventilated patients can decrease overall VAP rates. Among the available interventions, shortening the duration of mechanical ventilation and providing measures to prevent the aspiration of contaminated secretions are most important. Given the evidence supporting greater morbidity, hospital mortality, and medical care costs among patients who have VAP, the prevention of this nosocomial infection should be an important priority in the hospital setting. PMID- 15802176 TI - Nonresolving pneumonia. AB - Uncertainty over the expected clinical course of a community-acquired or nosocomial pneumonia is a common reason for pulmonary consultation. Determining which patients with prolonged pneumonia and at what point during therapy they should undergo further evaluation can be challenging. This article reviews "normal" resolution times for the most common pneumonias, risk factors for delayed resolution, and infectious and noninfectious conditions that can cause nonresolving pneumonia. An approach to the evaluation of the patient with this common problem is described. PMID- 15802177 TI - Neuronal expression of the NADPH oxidase NOX4, and its regulation in mouse experimental brain ischemia. AB - Ischemia-induced neuronal damage has been linked to elevated production of reactive oxygen species (ROS) both in animal models and in humans. NADPH oxidase enzymes (NOX-es) are a major enzymatic source of ROS, but their role in brain ischemia has not yet been investigated. The present study was carried out to examine the expression of NOX4, one of the new NADPH oxidase isoforms in a mouse model of focal permanent brain ischemia. We demonstrate that NOX4 is expressed in neurons using in situ hybridization and immunohistochemistry. Ischemia, induced by middle cerebral artery occlusion resulted in a dramatic increase in cortical NOX4 expression. Elevated NOX4 mRNA levels were detectable as early as 24 h after the onset of ischemia and persisted throughout the 30 days of follow-up period, reaching a maximum between days 7 and 15. The early onset suggests neuronal reaction, while the peak period corresponds to the time of neoangiogenesis occurring mainly in the peri-infarct region. The occurrence of NOX4 in the new capillaries was confirmed by immunohistochemical staining. In summary, our paper reports the presence of the ROS producing NADPH oxidase NOX4 in neurons and demonstrates an upregulation of its expression under ischemic conditions. Moreover, a role for NOX4 in ischemia/hypoxia-induced angiogenesis is suggested by its prominent expression in newly formed capillaries. PMID- 15802178 TI - delta-opioid receptor-mediated actions on rostral ventromedial medulla neurons. PMID- 15802179 TI - Daily restricted feeding rescues a rhythm of period2 expression in the arrhythmic suprachiasmatic nucleus. AB - Second only to light, daily restricted feeding schedules can entrain circadian rhythms in mammals [Neurosci Biobehav Rev 4 (1980) 119; J Biol Rhythms 17 (2002) 284]. Contrary to light, however, such feeding schedules have been found not to affect the master circadian clock in the suprachiasmatic nucleus (SCN) [Genes Dev 14 (2000) 2950; Eur J Neurosci 13 (2001) 1190]. Here, we show that in rats that are arrhythmic as a consequence of prolonged housing in constant light, a daily restricted feeding schedule not only restores behavioral rhythmicity, as previously shown [Physiol Behav 53 (1993) 509], but in addition, induces a rhythm of the clock protein, Period2 in the SCN. These findings challenge the idea that the SCN is invulnerable to feeding schedules and call for a reevaluation of the role of the SCN clock in the circadian effects of such schedules. PMID- 15802180 TI - Estrogen modifies stress response of catecholamine biosynthetic enzyme genes and cardiovascular system in ovariectomized female rats. AB - Estrogen is likely involved in the gender specific differences in coping with stress. Activation of catecholamine (CA) biosynthetic enzyme gene expression in central and peripheral CA systems plays a key role in response to stress and in regulation of the cardiovascular system. Here we examined whether estradiol can modulate response of hypothalamic-pituitary-adrenal axis (HPA), gene expression of enzymes related to CA biosynthesis in several noradrenergic locations, tetrahydrobiopterin (BH4) concentration and blood pressure (BP) in response to immobilization stress (IMO) of ovariectomized female rats. Rats were injected with 25 mug/kg estradiol benzoate (EB) or sesame oil once daily for 16 days and subsequently exposed to two hours of IMO. The IMO triggered elevation in plasma ACTH was lessened in EB-pretreated animals. However, estradiol did not alter the IMO-elicited rise of tyrosine hydroxylase mRNA levels in adrenal medulla (AM) and in the nucleus of solitary track (NTS) compared with controls. The response of GTP cyclohydrolase I (GTPCH) mRNA in AM to IMO was also similar in both groups. Several responses to IMO in EB-treated rats were reversed. Instead of IMO elicited elevation in dopamine beta-hydroxylase mRNA levels in the locus coeruleus, GTPCH mRNA and BH4 levels in the NTS, they were reduced by IMO. In a parallel experiment, BP was monitored during restraint stress. The elevation of BP in response to single or repeated restraint stress was sustained during 2 h in controls and reduced after 70 min stress in EB treated rats. One month after withdrawal of EB treatment, the BP response to restraint was similar to that of rats which never received EB. The results demonstrate that estrogen can modulate responses to stress affecting HPA axis, CA biosynthesis, in central and peripheral noradrenergic systems, and BP. PMID- 15802181 TI - Orexin-saporin lesions of the medial septum impair spatial memory. AB - The medial septum and diagonal band of Broca (MSDB) provide a major input to the hippocampus and are important for spatial learning and memory. Although electrolytic MSDB lesions have prominent memory impairing effects, selective lesions of either cholinergic or GABAergic MSDB neurons do not or only mildly impair spatial memory. MSDB neurons are targets of orexin-containing neurons from the hypothalamus. At present, the functional significance of orexin afferents to MSDB is unclear, and the present study investigated a possible involvement of orexin innervation of the MSDB in spatial memory. Orexin-saporin, a toxin that damages neurons containing the hypocretin-2 receptor, was administered into the MSDB of rats. Rats were subsequently tested on a water maze to assess spatial reference memory and a plus maze to assess spatial working memory. At 100 ng/microl, orexin-saporin destroyed primarily GABAergic septohippocampal neurons, sparing the majority of cholinergic neurons. At 200 ng/microl, orexin-saporin almost totally eliminated GABAergic septohippocampal neurons and destroyed many cholinergic neurons. Spatial reference memory was impaired at both concentrations of orexin-saporin with a dramatic impairment observed for 24-h retention. Short term reference memory was also impaired at both concentrations. Rats treated with 200 ng/microl, but not 100 ng/microl, of orexin-saporin were also impaired on a spontaneous alternation task, showing a deficit in spatial working memory. Our results, together with previous studies, suggest that orexin innervation of the MSDB may modulate spatial memory by acting on both GABAergic and cholinergic septohippocampal neurons. PMID- 15802182 TI - Learning twice is different from learning once and from learning more. AB - The rat hippocampus plays a crucial role in the consolidation of a variety of memories, including that for a one trial inhibitory avoidance learning task in which stepping down from a platform is associated with a footshock. Here we show that this is the case regardless of the intensity of the footshock used and hence, of the strength of the learned response. However, additional learning produced by a second training session in this task does not involve the hippocampus but, instead, the striatum. Memory consolidation of the second trial requires glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate, N methyl-D-aspartate and metabotropic receptors, activation of signaling pathways, gene expression and protein synthesis in the striatum, as are required in the hippocampus during memory consolidation of the first trial. PMID- 15802183 TI - The NR1-4 C-terminus interferes with N-methyl-D-aspartate receptor-mediated excitotoxicity: evidence against a typical T/SXV-PDZ interaction. AB - The N-methyl-D-aspartate receptor (NMDAR) plays a key role in the neural plasticity that underlies learning and memory in vivo. The plasticity exhibited by NMDARs may also contribute to disease pathogenesis, as a number of disorders are caused or exacerbated by exaggerated NMDAR activity. The NMDAR is composed of two obligatory types of subunits, NR1 and NR2. These transmembrane proteins include large intracellular C-termini that have yet to be fully characterized. We have developed a three-color fluorescence system in order to visualize NMDAR expression in living cells. Using excitotoxicity as a proxy for exaggerated NMDAR activity, we analyzed the effect of over-expressing NR1-4 and NR2A C-terminal domains on exaggerated NMDAR function. We demonstrate that a determinant within the C-terminal domain of NR1-4 (C02') is important for NMDAR excitotoxicity, whereas no novel determinants were identified in the NR2A C-terminus. Through the use of heterologous cells, and by examining the interaction between the prototypical NMDAR-binding partner postsynaptic density-95 (PSD-95), we show that this effect is unlikely to be mediated through a classical interaction with PSD 95. PMID- 15802184 TI - 17beta-estradiol induces Ca2+ influx, dendritic and nuclear Ca2+ rise and subsequent cyclic AMP response element-binding protein activation in hippocampal neurons: a potential initiation mechanism for estrogen neurotrophism. AB - Previous studies from our laboratory have shown that 17beta-estradiol (E2) promotes neurite outgrowth in hippocampal and cortical neurons. The neurotrophic effect of E2 seen in vitro has also been observed in vivo by other investigators who found that E2 enhances the density of dendritic spines involved in neuronal synaptic connection. To investigate the rapid upstream mechanisms initiating the E2 neurotrophic effect, we tested the hypothesis that E2 would directly activate Ca2+ influx in primary hippocampal neurons, which would result in activation of the transcription factor, cyclic AMP response element-binding protein (CREB), and regulate E2 enhancement of neurite outgrowth. Using fura-2 ratiometric and fluo-3 Ca2+ imaging, we demonstrated that E2 induced a significant rise in intracellular Ca2+ concentration ([Ca2+]i) through E2-induced Ca2+ influx. Interestingly, the rise in [Ca2+]i occurred not only in the cytoplasm, but also in the nucleus and dendrites of hippocampal neurons. Since CREB is activated by Ca2+-dependent kinases and is required for certain aspects of synaptic plasticity, we investigated whether E2 would lead to activation of CREB. Western immunoblotting and immunocytochemical analyses revealed that E2 induced rapid CREB activation consistent with rapid intracellular Ca2+ signaling, which was dependent on the influx of extracellular Ca2+. E2-induced increase in dendritic spine marker protein spinophilin was abolished following treatment with a small interfering RNA against CREB, indicating that E2-induced neurotrophic effect requires the upstream CREB activation. Results of these analyses indicate that E2-induced neurotrophic responses are mediated by a Ca2+ signaling cascade that is dependent upon extracellular Ca2+ and CREB activation. These data provide insights into the initiating mechanisms required to activate the estrogen neurotrophic response and provide a mechanistic framework for determining the neurotrophic efficacy of existing and emerging estrogen therapies for the brain. PMID- 15802185 TI - Proteomic identification of proteins specifically oxidized by intracerebral injection of amyloid beta-peptide (1-42) into rat brain: implications for Alzheimer's disease. AB - Protein oxidation has been shown to result in loss of protein function. There is increasing evidence that protein oxidation plays a role in the pathogenesis of Alzheimer's disease (AD). Amyloid beta-peptide (1-42) [Abeta(1-42)] has been implicated as a mediator of oxidative stress in AD. Additionally, Abeta(1-42) has been shown to induce cholinergic dysfunction when injected into rat brain, a finding consistent with cholinergic deficits documented in AD. In this study, we used proteomic techniques to examine the regional in vivo protein oxidation induced by Abeta(1-42) injected into the nucleus basalis magnocellularis (NBM) of rat brain compared with saline-injected control at 7 days post-injection. In the cortex, we identified glutamine synthetase and tubulin beta chain 15/alpha, while, in the NBM, we identified 14-3-3 zeta and chaperonin 60 (HSP60) as significantly oxidized. Extensive oxidation was detected in the hippocampus where we identified 14-3-3 zeta, beta-synuclein, pyruvate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase, and phosphoglycerate mutase 1. The results of this study suggest that a single injection of Abeta(1-42) into NBM can have profound effects elsewhere in the brain. The results further suggest that Abeta(1-42) induced oxidative stress in rat brain mirrors some of those proteins oxidized in AD brain and leads to oxidized proteins, which when inserted into their respective biochemical pathways yields insight into brain dysfunction that can lead to neurodegeneration in AD. PMID- 15802186 TI - Effects of postnatal nicotine exposure on apoptotic markers in the developing piglet brain. AB - Exposure to cigarette smoke is a risk factor for the sudden infant death syndrome (SIDS), but the ability to distinguish between the neuropathological effects of pre- versus postnatal exposure is limited in the clinical setting. To test whether postnatal nicotine exposure could contribute to the increased neuronal expression of apoptotic markers that we have previously observed in SIDS infants, as well as including study of gender influences, we developed a piglet model to mimic passive smoking in the early postnatal period. Piglets were exposed to nicotine (2 mg/kg/day infused via an implanted osmotic minipump) within 48 h of birth until the age of 13-14 days, when the brain was collected for study. Four piglet groups included: control females (n=7), control males (n=7), nicotine females (n=7), and nicotine males (n=7). Apoptotic markers included immunohistochemistry for activated caspase-3, and for DNA fragmentation or terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) in seven nuclei of the brainstem caudal medulla and two subregions of the hippocampus (CA4 and dentate gyrus). Among control females compared with males, there was less active caspase-3 and less TUNEL in the dorsal motor nucleus of vagus (DMNV), and there was less TUNEL in the nucleus of the spinal trigeminal tract (NSTT). Compared with controls, nicotine-exposed male piglets had increased TUNEL staining in the cuneate nucleus (P=0.05), and increased active caspase-3 in the hypoglossal, gracile and dentate gyrus (P<0.05 for each). Nicotine-exposed females showed no change in TUNEL staining in any of the nuclei studied, but increased active caspase-3 in the hypoglossal, DMNV and NSTT (P<0.05 for each). These results show for the first time that postnatal nicotine exposure can lead to an increase in apoptotic markers in the brain. In piglets, these effects showed regional and gender-specific differences, suggesting that passive, postnatal nicotine exposure may be responsible for some neuropathological changes observed in infants dying from SIDS. PMID- 15802187 TI - Antiepileptic action induced by a combination of vigabatrin and tiagabine. AB - Vigabatrin, an inhibitor of GABA breakdown by GABA transaminase and of GABA transporter isoform 1 (GAT1), and tiagabine, a highly specific inhibitor of GAT1, have successfully been applied in the treatment of epilepsy. We investigated the effects of individual and combined application of these drugs on GAT1 expressed in Xenopus oocytes, and examined the effects on epileptiform discharges in the CA3 area of brain slices of genetically epileptic El and control ddY mice, and on the occurrence of seizures in El mice. Simultaneous application of vigabatrin and tiagabine inhibited epileptiform discharges induced by high-K+ solution in the brain slices in an antagonistic fashion. The degree of inhibition by tiagabine after pre-treatment with vigabatrin was additive in ddY mice and synergistic in El mice. In Mg2+-free solution, co-treatment by the two drugs produced additive inhibition in slices from both mouse strains, but pre-treatment with vigabatrin produced synergistic inhibition in slices only from ddY mice. In the slices from El mice, a combination of drugs resulted in additive effects in both co- and pre treatment by the drugs. Although these drugs are also effective in vivo at suppressing seizure occurrence in El mice, the combined application does not show synergistic effects, but rather is antagonistic under the experimental conditions in this particular variant of epilepsy. The synergistic inhibition of epileptiform discharges in brain slices may, in part, have originated from the complex interaction with GAT1. In experiments on the GAT1 expressed in oocytes it could be demonstrated that synergistic inhibition occurs only at low concentration (0.1 nM) of vigabatrin. This illustrates that the oocytes may form a powerful test system for drug screening and investigation of complex drug interactions. These results present a novel interpretation of synergistic inhibition of certain epileptic discharges using vigabatrin and another drug, and that for successful synergistic treatment of epilepsies carefully designed timed dosage regimens are essential. PMID- 15802188 TI - Prefrontal cortical modulation of acetylcholine release in posterior parietal cortex. AB - Attentional processing is a crucial early stage in cognition and is subject to "top-down" regulation by prefrontal cortex (PFC). Top-down regulation involves modification of input processing in cortical and subcortical areas, including the posterior parietal cortex (PPC). Cortical cholinergic inputs, originating from the basal forebrain cholinergic system, have been demonstrated to mediate important aspects of attentional processing. The present study investigated the ability of cholinergic and glutamatergic transmission within PFC to regulate acetylcholine (ACh) release in PPC. The first set of experiments demonstrated increases in ACh efflux in PPC following AMPA administration into the PFC. These increases were antagonized by co-administration of the AMPA receptor antagonist DNQX into the PFC. The second set of experiments demonstrated that administration of carbachol, but not nicotine, into the PFC also increased ACh efflux in PPC. The effects of carbachol were attenuated by co-administration (into PFC) of a muscarinic antagonist (atropine) and partially attenuated by the nicotine antagonist mecamylamine and DNQX. Perfusion of carbachol, nicotine, or AMPA into the PPC did not affect PFC ACh efflux, suggesting that these cortical interactions are not bi-directional. These studies demonstrate the capacity of the PFC to regulate ACh release in the PPC via glutamatergic and cholinergic prefrontal mechanisms. Prefrontal regulation of ACh release elsewhere in the cortex is hypothesized to contribute to the cognitive optimization of input processing. PMID- 15802189 TI - Cocaine administration produces a protracted decoupling of neural and haemodynamic responses to intense sensory stimuli. AB - Evidence suggests that for relatively weak sensory stimuli, cocaine elevates background haemodynamic parameters but still allows enhanced neural responses to be reflected in enhanced haemodynamic responses. The current study investigated the possibility that for more intense stimuli, the raised background may produce a protracted attenuation of the haemodynamic response. Three experiments were performed to measure effects of i.v. cocaine administration (0.5 mg/kg) or saline on responses in rat barrel cortex to electrical stimulation of the whisker pad. The first experiment used optical imaging spectroscopy (OIS) and laser Doppler flowmetry (LDF) to measure haemodynamic changes. Cocaine caused an increase in baseline blood flow (peak approximately 90%), which lasted for the duration of the test period (25 min). Haemodynamic responses to whisker stimulation were substantially reduced throughout. The second experiment used a 16-channel multi electrode to measure evoked potentials at 100 mum intervals through the barrel cortex. Summed neural responses (collapsed across the spatial dimension) after cocaine administration were similar to those after saline. The third experiment extended experiment 1 by examining the effects of cocaine on whisker sensory responses using functional magnetic resonance imaging (and concurrent OIS or LDF). Cocaine caused a similar increase in baseline and reduction in the evoked response to that seen in experiment 1. Together, the results of these three experiments show that cocaine produces a protracted decoupling of neural activity and haemodynamic responses to intense sensory stimulation, which suggests that imaging techniques based on changes in haemodynamic parameters may be unsuitable for studying the effects of cocaine on sensory processing in humans. PMID- 15802190 TI - Sex differences in the stereological parameters of the hippocampal dentate gyrus of the guinea-pig before puberty. AB - Studies in rats and mice have shown several sex-dependent functional and structural differences in the hippocampal region, a brain structure playing a key role in learning and memory. The aim of the present study was to establish whether sex differences exist prior to puberty in the stereological parameters of the dentate gyrus in the guinea-pig, a long-gestation rodent, whose brain is at a more advanced stage of maturation at birth than the rat and mouse. The number of granule cells and volumes of the granule cell layer, molecular layer and hilus were evaluated in Nissl-stained brains of neonatal (15-16 days old) and peripubescent (45-46 days old) guinea-pigs. Based on a pilot study, the optical disector method was preferred to the optical fractionator method to estimate cell number. For volume (Vref) estimation with the Cavalieri principle, contour tracing was preferred to the point counting method, as the latter appeared to underestimate volumes. The results showed that neonatal males had more granule cells than females in both the dorsal and ventral dentate gyrus and a larger volume in all layers. Peripubescent males had a larger volume of the granule cell layer than females in both the dorsal and ventral dentate gyrus, more granule cells in the ventral dentate gyrus, a larger volume of the hilus in both the dorsal and ventral dentate gyrus and a larger volume of the molecular layer in the ventral dentate gyrus. The results show that sex differences are present in the guinea-pig dentate gyrus prior to puberty and go in the same direction at both investigated ages, with males exhibiting more granule cells and larger volumes than females. The widespread distribution of these sex differences suggests that in the guinea-pig, similarly to other rodents, hippocampus dependent functions may be sexually dimorphic. PMID- 15802191 TI - Smoking during early pregnancy affects the expression pattern of both nicotinic and muscarinic acetylcholine receptors in human first trimester brainstem and cerebellum. AB - Prenatal nicotine exposure is associated with an increased risk of complications during pregnancy and childhood. In this study the expression of nicotinic and muscarinic acetylcholine receptors in first trimester pons, medulla oblongata and cerebellum from abortus (5-12 weeks of gestation) of smoking and nonsmoking women was compared. A significant age-related increase in binding of nicotinic receptor subtype alpha4 was found in both pons and cerebellum only in fetal tissue from non-smoking women, while a similar increase was observed in medulla oblongata from fetuses exposed to smoking. A significant age-related increase in binding of muscarinic receptor subtype m2 was observed in pons from abortus of smoking compared with non-smoking women. The gene expression pattern of both alpha4 and alpha7 nicotinic receptor subunits was changed after smoking in all three regions investigated. Smoking also changed the expression of m1 and 2 muscarinic receptor mRNA in pons, m1 mRNA in cerebellum and the m3 mRNA in medulla oblongata. The findings indicate that early prenatal nicotine exposure affects the normal developmental pattern of the cholinergic system in human fetal brain. PMID- 15802192 TI - Long-term effects of diazepam and phenobarbital treatment during development on GABA receptors, transporters and glutamic acid decarboxylase. AB - Diazepam (DZ) and phenobarbital (PH) are commonly used to treat early-life seizures and act on GABAA receptors (GABAR). The developing GABAergic system is highly plastic, and the long-term effects of postnatal treatment with these drugs on the GABAergic system has not been extensively examined. In the present study, we investigated the effects of prolonged DZ and PH treatment during postnatal development and then discontinuation on expression of a variety of genes involved in GABAergic neurotransmission during adulthood. Rat pups were treated with DZ, PH or vehicle from postnatal day (P) 10-P40 and then the dose was tapered for 2 weeks and terminated at P55. Expression of GABAR subunits, GABAB receptor subunits, GABA transporters (GAT) and GABA synthesizing enzymes (glutamic acid decarboxylase: GAD) mRNAs in hippocampal dentate granule neurons (DGNs) were analyzed using antisense RNA amplification at P90. Protein levels for the alpha1 subunit of GABAR, GAD67, GAT1 and 3 were also assessed using Western blotting. At P90, mRNA expression for GAT-1, 3, 4, GABAR subunits alpha4, alpha6, beta3, delta and theta and GABAB receptor subunit R1 was increased and mRNA expression for GAD65, GAD67 and GABAR subunits alpha1 and alpha3 were decreased in DGNs of rats treated with DZ and PH. The current data suggest that prolonged DZ and PH treatment during postnatal development causes permanent alterations in the expression of hippocampal GABA receptor subunits, GATs and GAD long after therapy has ended. PMID- 15802194 TI - Evidence that projections from the bed nucleus of the stria terminalis and from the lateral and medial regions of the preoptic area provide input to gonadotropin releasing hormone (GNRH) neurons in the female sheep brain. AB - The arcuate nucleus/ventromedial hypothalamic nucleus (ARC/VMH) region is thought to relay estrogen feedback signals to gonadotropin-releasing hormone (GnRH) cells in the sheep brain. This region sends major projections to the lateral preoptic area (lPOA), ventral bed nucleus of the stria terminals (vBnST) and the ventro caudal division of the median preoptic nucleus (vcMePON) with little direct input to GnRH cell bodies, suggesting interneuronal relay to GnRH neurons. The brain stem also provides input to the POA. The present study aimed to identify possible relay circuits in the POA and BnST to GnRH neurons. Biotinylated dextran amine (BDA) was injected into lPOA (n=6), vBnST (n=2), vcMePON (n=3) and periventricular nucleus (PeriV; n=1) of ewes for anterograde tracing. GnRH immunoreactive (IR) perikarya appearing to receive input from BDA-containing varicosities were identified by fluorescence microscopy, with further analysis by confocal microscopy. When BDA was injected into rostral and caudal regions of lPOA (n=3), no tracer-filled varicose fibers were found in contact with GnRH-IR perikarya. Injections into the center of the lPOA (n=3) indicated direct projections to GnRH-IR cells. Injections into the vBnST, vcMePON and PeriV indicated that cells of these regions also provide input to GnRH cells. BDA containing varicosities found in the MPOA were immunoreactive for NPY or were GABAergic or glutamatergic when the tracer was injected into vBnST and lPOA, but not when injections were placed in the vcMePON. With injection into the PeriV, tracer-filled varicosities in the MPOA were not immunoreactive for somatostatin or enkephalin. Injection of FluoroGold into ventral POA retrogradely labeled cells in the above mentioned areas, but few were also immunoreactive for estrogen receptor-alpha. Thus, cells of the vBnST, lPOA, vcMePON and PeriV project to GnRH neurons. These cells may provide an interneuronal route to GnRH neurons from the ARC/VMH, the brain stem and other regions of the brain. PMID- 15802193 TI - Enhanced striatal opioid receptor-mediated G-protein activation in L-DOPA-treated dyskinetic monkeys. AB - Long-term l-3,4-dihydroxyphenylalanine (L-DOPA) treatment in Parkinson's disease leads to dyskinesias in the majority of patients. The underlying molecular mechanisms for L-DOPA-induced dyskinesias (LIDs) are currently unclear. However, the findings that there are alterations in opioid peptide mRNA and protein expression and that opioid ligands modulate dyskinesias suggest that the opioid system may be involved. To further understand its role in dyskinesias, we mapped opioid receptor-stimulated G-protein activation using [35S]guanylyl-5'-O-(gamma thio)-triphosphate ([35S]GTPgammaS) autoradiography in the basal ganglia of normal and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned squirrel monkeys administered water or L-DOPA. Subtype-selective opioid receptor G-protein coupling was investigated using the mu-opioid agonist [D-Ala, N-Me-Phe, Gly-ol] enkephalin, delta-agonist SNC80 and kappa-agonist U50488H. Our data show that mu opioid receptor-mediated G-protein activation is significantly enhanced in the basal ganglia and cortex of L-DOPA-treated dyskinetic monkeys, whereas delta- and kappa-receptor-induced increases were limited to only a few regions. A similar pattern of enhancement was observed in both MPTP-lesioned and unlesioned animals with LIDs suggesting the effect was not simply due to a compromised nigrostriatal system. Opioid receptor G-protein coupling was not enhanced in non-dyskinetic L DOPA-treated animals, or lesioned monkeys not given L-DOPA. The increases in opioid-stimulated [35S]GTPgammaS binding are directly correlated with dyskinesias. The present data demonstrate an enhanced subtype-selective opioid receptor G-protein coupling in the basal ganglia of monkeys with LIDs. The positive correlation with LIDs suggests this may represent an intracellular signaling mechanism underlying these movement abnormalities. PMID- 15802195 TI - Biophysical characterization of whole-cell currents in O2-sensitive neurons from the rat glossopharyngeal nerve. AB - In this study we use nystatin perforated-patch and conventional whole-cell recording to characterize the biophysical properties of neuronal nitric oxide synthase (nNOS)-expressing paraganglion neurons from the rat glossopharyngeal nerve (GPN), that are thought to provide NO-mediated efferent inhibition of carotid body chemoreceptors. These GPN neurons occur in two populations, a proximal one near the bifurcation of the GPN and the carotid sinus nerve, and a more distal one located further along the GPN. Both populations were visualized in whole mounts by vital staining with the styryl pyridinium dye, 4-Di-2-ASP (D289). Following isolation in vitro, proximal and distal neurons had similar input resistances (mean: 1.5 and 1.6 GOmega, respectively), input capacitances (mean: 25.0 and 27.4 pF, respectively), and resting potentials (mean: -53.9 and 53.3 mV, respectively). All neurons had similar voltage-dependent currents composed of: tetrodotoxin (TTX)-sensitive Na+ currents (IC50 approximately 0.2 microM), prolonged and transient Ca2+ currents, and delayed rectifier-type K+ currents. Threshold activation for the Na+ currents was approximately -30 mV and they were inactivated within 10 ms. Inward Ca2+ currents consisted of nifedipine sensitive L-type, omega-agatoxin IVA-sensitive P/Q-type, omega-conotoxin GVIA sensitive N-type, SNX-482-sensitive R-type, and Ni2+-sensitive, but SNX-482 insensitive, T-type channels. The voltage-dependent outward K+ currents were sensitive to tetraethylammonium (TEA; 10 mM) and 4-aminopyridine (4-AP; 2 mM). Exposure to a chemosensory stimulus, hypoxia (PO2 range: 80-5 Torr), caused a dose-dependent decrease in K+ current which persisted in the presence of TEA and 4-AP, consistent with the involvement of background K+ channels. Under current clamp, GPN neurons generated TTX-sensitive action potentials, and in spontaneously active neurons, hypoxia caused membrane depolarization and an increase in firing frequency. These properties endow GPN neurons with an exquisite ability to regulate carotid body chemoreceptor function during hypoxia, via voltage-gated Ca2+-entry, activation of nNOS, and release of NO. PMID- 15802196 TI - A new role for the renin-angiotensin system in the rat periaqueductal gray matter: angiotensin receptor-mediated modulation of nociception. AB - Renin-angiotensin (Ang) system (RAS) peptides injected into the periaqueductal gray matter (PAG) elicit antinociception. Saralasin blocks Ang II-elicited antinociception. Thus, it is possible that endogenous RAS peptides could participate on the modulation of nociception in the PAG. This possibility was tested here injecting, in the PAG, the specific Ang type 1 and type 2 receptor (AT1 receptor and AT(2 receptor) antagonists losartan and CGP42,112A, respectively, either alone or before Ang II. The effects of Ang II, losartan and CGP42,112A on nociception were measured using the tail flick test and the model of incision allodynia. Ang II increased tail-flick latency, an effect inhibited by both losartan and CGP42,112A. Ang II reduced incisional allodynia. Either losartan or CGP42,112A alone increased incision allodynia, suggesting that endogenous Ang II and/or an Ang-peptide participates in the control of allodynia by the PAG. AT1 and AT2 receptors were immunolocalized in neuronal cell bodies and processes in the ventrolateral PAG. Taken together, the antinociceptive effect of Ang II injection into the ventrolateral PAG, the increase of allodynia elicited by injecting either losartan or CGP42,112A alone in the PAG, and the presence of AT1 and AT2 receptors in neurons and neuronal processes in the same region, represent the first evidence that part of the tonic nociceptive control mediated by the PAG is carried out locally by endogenous Ang II and/or an Ang peptide acting on AT1 and AT2 receptors. PMID- 15802197 TI - Environmental light and suprachiasmatic nucleus interact in the regulation of body temperature. AB - The mammalian biological clock, located in the suprachiasmatic nucleus (SCN), is crucial for circadian rhythms in physiology and behavior. However, equivocal findings have been reported on its role in the circadian regulation of body temperature. The goal of the present studies was to investigate the interaction between the SCN and environmental light in the regulation of body temperature. All recordings were performed by telemetry in free moving male Wistar rats. Firstly, we demonstrated an endogenous circadian rhythm in body temperature independent of locomotor activity. This rhythm was abolished by stereotactic lesioning of the SCN. Secondly, we demonstrated a circadian phase-dependent suppressive effect of light ('negative masking') on body temperature. Light suppressed body temperature more at the end of the subjective night (circadian time [CT] 22) than in the middle (CT 6) and at the end (CT 10) of the subjective day. This circadian-phase dependent suppression was not demonstrated in SCN lesioned animals. Surprisingly, after half a year of recovery from lesioning of the SCN, light regained its suppressing action on body temperature, resulting in a daily body temperature rhythm only under light-dark conditions. In contrast to body temperature, light could not substantially mimic a daytime inhibitory SCN output in the regulation of heart rate and locomotor activity. The present results suggest that, after lesioning of the SCN as main relay station for the immediate body temperature-inhibition by light, secondary relay nuclei can fully take over this function of the SCN. These findings provide a possible explanation for the controversy in literature over the question whether the SCN is required for the diurnal rhythm in body temperature. Furthermore, they show that light may have an acute effect on behavior and physiology of the organism via the SCN, which extends beyond the generally acknowledged effect on melatonin secretion. PMID- 15802198 TI - Nociceptive characteristics of tumor necrosis factor-alpha in naive and tumor bearing mice. AB - A nociceptive role for tumor necrosis factor-alpha (TNF-alpha) in naive mice and in mice with fibrosarcoma tumor-induced primary hyperalgesia was investigated. The presence of TNF-alpha mRNA was confirmed in tumor site homogenates by reverse transcription-polymerase chain reaction (RT-PCR), and examination of TNF-alpha protein levels in tumor-bearing mice indicated a significantly higher concentration of this cytokine in tumor microperfusates and tumor site homogenates compared with that obtained from a similar site on the contralateral limb or in naive mice. Intraplantar injection of TNF-alpha into naive or fibrosarcoma tumor-bearing mice induced mechanical hypersensitivity, as measured by withdrawal responses evoked by von Frey monofilaments. This hypersensitivity suggests that TNF-alpha can excite or sensitize primary afferent fibers to mechanical stimulation in both naive and tumor-bearing mice. In addition, the hyperalgesia produced by TNF-alpha was completely eliminated when the injected TNF-alpha was pre-incubated with the soluble receptor antagonist TNFR:Fc. Importantly, pre-implantation systemic as well as post-implantation intra-tumor injection of TNFR:Fc partially blocked the mechanical hyperalgesia, indicating that local production of TNF-alpha may contribute to tumor-induced nociception. PMID- 15802199 TI - Type IX collagen is crucial for normal hearing. AB - cDNA microarray analysis indicated that COL9A1 and COL9A3 are highly expressed in the human inner ear, suggesting that type IX collagen has a crucial functional role in the inner ear. This study further confirmed, by means of real-time PCR, the presence of collagen type IX genes in the mouse inner ear. Immunocytochemical analysis also revealed that type IX collagen is distributed in the tectorial membrane, where it co-localizes with type II collagen, indicating that type IX collagen may contribute to the three-dimensional integrated structure of type II collagen molecules. Mice with targeted disruption of the col9a1 gene were shown through assessment by auditory brain stem response to have hearing loss, suggesting an important role of type IX collagen in maintaining normal hearing. At the light microscopic level, the tectorial membrane of knock-out mice was found to be abnormal in shape, and electron microscopy confirmed disturbance of organization of the collagen fibrils. An antibody against type II collagen failed to detect type II collagen in the tectorial membrane of type IX collagen knock out mice, suggesting that a lack of type IX collagen may affect the three dimensional structure of type II collagen molecules. These findings indicate that genes encoding each chain of type IX collagen may fulfill an important function associated with the tectorial membrane in the auditory system. PMID- 15802200 TI - Acetylcholine release is elicited in the visual cortex, but not in the prefrontal cortex, by patterned visual stimulation: a dual in vivo microdialysis study with functional correlates in the rat brain. AB - By its projections to the primary visual and the prefrontal cortices, the basal forebrain cholinergic system is involved in cognitive processing of sensory stimuli. It has been suggested that visual stimulus-induced cholinergic activation of the visual cortex may exert a permissive role on thalamocortical inputs. However, it is not known if visual stimulation elicits cholinergic activation of high-order brain areas in the absence of attentional need. In the present study, we measured the effects of patterned visual stimulation (horizontal grating) on the release of acetylcholine with dual-probe in vivo microdialysis in the visual and the prefrontal cortices of anesthetized rats. We also used retrograde tracing to determine the anatomical relationships of cholinergic neurons with neurons of the visual system and the prefrontal cortex. Finally, we evaluated a functional correlate of this stimulation, namely c-fos immunolabeling. Patterned visual stimulation elicited significant increases in acetylcholine release in the visual cortex, accompanied by an increased number of c-fos immunoreactive neurons in this brain area. In contrast, in the prefrontal cortex, neither the level of acetylcholine release nor the number of c-fos immunoreactive neurons was significantly changed because of the stimulation. Cholinergic basal forebrain neurons projecting to the visual or the prefrontal cortices were both localized within the horizontal limb of the diagonal band of Broca but were not immunoreactive for c-fos during visual stimulation. No parts of the visual system were found to directly project to these basal forebrain neurons. These results suggest the differential involvement of cholinergic projections in the integration of sensory stimuli, depending on the level of activity of the targeted cortical area. PMID- 15802201 TI - Fos-related antigen 2 (Fra-2) memorizes photoperiod in the rat pineal gland. AB - As the physiological role of fos-related antigen-2 (Fra-2) is largely unknown and since the pineal plays an important role in the photoperiodic control of the body, we have tested the hypothesis that Fra-2 expression is photoperiod dependent and may be involved in imprinting photoperiod on the pineal gland and the body as a whole. To this end, we have investigated Fra-2 mRNA expression and Fra-2 protein expression under various light/dark (LD) cycles. A clear nocturnal increase occurs for both monitored parameters under all photoperiodic conditions studied. The level of Fra-2 protein expression clearly depends on photoperiod, because the amount of protein at dark onset and during the night negatively correlates with the length of the photoperiod. Further, high-phosphorylated Fra-2 isoforms are abundant under all photoperiods tested, with the exception of LD 20:4. Because Fra-2 phosphorylation depends on cGMP, a depressed cGMP response to adrenergic stimulation under LD 20:4 appears to explain this finding. We conclude that photoperiod is imprinted on Fra-2 in terms of both protein amount and protein phosphorylation in the rat pineal gland. This imprinting becomes fully manifest after about 7 days only, suggesting that a number of altered photoperiodic cycles are required for pineal Fra-2 to "learn" that the photoperiod has changed. Reportedly, Fra-2 limits expression of the enzyme iodothyronine deiodinase type II, which catalyzes the intracellular deiodination of thyroxine prohormone to the active 3,3',5-triiodothyronine. We have found that the extent of Fra-2 expression inversely correlates with the dII gene response to cAMP; hence the photoperiodic regulation of Fra-2 may affect the body by changing pineal thyroid hormone metabolism. PMID- 15802202 TI - Circadian firing-rate rhythms and light responses of rat habenular nucleus neurons in vivo and in vitro. AB - The suprachiasmatic nuclei of the anterior hypothalamus serve as the principal pacemaker of the mammalian circadian system. Among its efferent targets are the habenular nucleus (Hb), especially the lateral Hb (LHb), which plays an important role in conveying input from the limbic forebrain to midbrain structures. We recorded extracellularly from single neurons in the LHb and medial Hb (MHb), both in vivo and using an in vitro slice preparation, to assess their responses to retinal illumination and the rhythmicity of their firing rates. Of cells recorded in the LHb, 42% were tonically activated or suppressed by retinal illumination, while significantly fewer cells recorded in the MHb responded to retinal illumination (19%). Of photically responsive cells, 68% in the LHb were activated and the remainder suppressed, while only 25% of those recorded in the MHb were activated. Cells in both the LHb and MHb showed higher baseline firing rates during the day than during the night in vivo, while photic responses were of significantly larger amplitude among LHb cells during the projected night than during the projected day. LHb cells recorded in vitro maintained their rhythmicity for two circadian cycles, but MHb cells did not show a rhythm in vitro. The habenula may play a role in linking circadian and motivational systems and may contribute to photic regulation of these systems, as well as to the rhythmicity of their function. PMID- 15802203 TI - Overnight verbal memory retention correlates with the number of sleep spindles. AB - Despite strong evidence supporting a role for sleep in the consolidation of newly acquired declarative memories, the contribution of specific sleep stages remains controversial. Based on electrophysiological studies in animals, synchronous sleep oscillations have been long proposed as possible origins of sleep-related memory improvement. Nevertheless, no studies to date have directly investigated the impact of sleep oscillations on overnight memory retention in humans. In the present study we provide evidence that overnight verbal memory retention is highly correlated with the number of sleep spindles detected by an automatic algorithm over left frontocentral areas. At the same time, overnight retention of newly learned faces was found to be independent of spindle activity but correlated with non-rapid-eye-movement sleep time. The data strongly support theories suggesting a link between sleep spindle activity and verbal memory consolidation. PMID- 15802204 TI - Sex and housing conditions affect the 24-h acetylcholine release profile in the hippocampus in rats. AB - To examine the sex difference in the 24-h profile of the acetylcholine (ACh) release in the hippocampus, in addition to the effects of housing conditions on this profile, we performed an in vivo microdialysis study in intact male and cycling female rats that had been living in large (diameter=35 cm) or small (diameter=19 cm) cylindrical cages. Each rat was individually housed in a cage for 4 days. On the day of the experiment, the dialysate was collected from the dorsal hippocampus at 20-min intervals and sequential blood samples were simultaneously obtained at 2-h intervals, under the freely moving condition for more than 24 h. ACh in the dialysates was measured by the high performance liquid chromatography system, while the corticosterone concentration in the serum was measured by radiostereoassay. Although the ACh release showed a clear daily rhythm in both sexes of rats, the amount of ACh released in female rats was significantly lower than that in males. Furthermore, the housing in the small cage significantly attenuated the ACh release during the dark phase in male rats, but not in female rats. Conversely, the serum corticosterone concentration showed a clear daily rhythm and the mean concentration of serum corticosterone in female rats was significantly higher than that in male rats. Housing in the small cage did not affect the corticosterone rhythm in either sex. These results reveal a sex difference in the 24-h profile of the ACh release, which suggests vulnerability of the cholinergic system in male rats depending on its housing conditions. PMID- 15802205 TI - Life-threatening skin diseases: part I. PMID- 15802206 TI - Life-threatening cutaneous conditions in neonates. AB - One of the major medical concerns in neonates is their vulnerability to life threatening complications. Early diagnosis of potentially serious medical conditions is essential in prevention of such complication by prompt administration of appropriate treatments. Recognition of cutaneous manifestations of potentially fatal diseases may assist a clinician in management of newborns. To overview the full spectrum of this category of conditions, Medline search was performed for cases of cutaneous manifestations associated with mortality during the first month of live that were published in the English language literature since 1977. This is a concise review of the clinical aspects of most of the potentially life-threatening disorders in neonates divided by basic morphology of their cutaneous manifestations. PMID- 15802207 TI - Life-threatening necrotizing fasciitis. AB - Necrotizing fasciitis is characterized by a rapidly progressing necrosis of subcutaneous fat and fascia, which can be life-threatening without prompt recognition, surgical intervention, and immediate antibiotic therapy. Necrotizing fasciitis has been subdivided into type 1, or polymicrobial necrotizing fasciitis, and type 2, or group A streptococcal necrotizing fasciitis. In addition, synonyms, such as streptococcal gangrene and "flesh-eating bacteria syndrome," have been used in the literature. PMID- 15802208 TI - Life-threatening eruptions due to infectious agents. AB - Some infectious diseases may cause rapidly fatal eruptions that need to be diagnosed and treated in an early phase for patient survival. The main life threatening eruptions of infectious etiology include Rocky Mountain spotted fever, meningococcemia, toxic shock syndrome, streptococcal toxic shock syndrome, and staphylococcal scalded skin syndrome. PMID- 15802209 TI - Life-threatening cutaneous viral diseases. AB - Most viral infections present with a cutaneous eruption that in the greater majority of cases is self-healing. Only in rare instances is the eruption accompanied by constitutional symptoms, and even more rarely, irreversible damage may occur or death may ensue. As with other infectious diseases, prognosis depends on the status of immunologic surveillance. Immunodepressed patients are, therefore, at the greatest risk. It has become more evident, however, that even in immunocompetent patients, eruption that were considered not long ago as benign diseases may in fact result in complications and even death. The introduction of polymerase chain reaction assays has allowed us to attribute conditions to a specific viral infection, which in the past was not possible. Therefore, dermatologists may come across, directly or during consultation, patients presenting with skin lesions that may be related to a viral disease and may lead to possible severe systemic complications. Dermatologists, however, may not be prepared for such developments, and thus the goal of this article is to suggest some of the possibilities. Discussing all the viral diseases that may present with cutaneous manifestations would be cumbersome; thus this article will be limited to the diseases that may endanger the life of a patient who is seeking dermatologic advice and who does not seem to have, at first sight, severe systemic involvement. PMID- 15802210 TI - Life-threatening stings, bites, infestations, and parasitic diseases. AB - Bites, stings and infestations can be fatal. Anaphylaxis to vespids and bees can be prevented with immunotherapy. Patients should be referred to an allergist. The acute care and prevention of arthropod injury is discussed below. PMID- 15802211 TI - Life-threatening acute adverse cutaneous drug reactions. AB - Adverse cutaneous reactions to drugs are frequent, affecting 2% to 3% of all hospitalized patients. Fortunately, only about 2% of adverse cutaneous reactions are severe and very few are fatal. Stevens-Johnson syndrome and toxic epidermal necrolysis are severe life-threatening diseases with a mortality rate reaching 30%, and only prompt recognition and diagnosis, withdrawal of the offensive drug, and referral to an intensive care unit or burn care unit might improve the prognosis and save the patient's life. Drug eruption with eosinophilia and systemic symptoms syndrome, formerly termed drug hypersensitivity syndrome, is a rather distinct severe adverse drug reaction (ADR) characterized by eruption, fever, lymph node enlargement, and single or multiple organ involvement, with a high morbidity and a mortality rate of 10%. These severe ADRs, together with serum sickness-like syndrome, are discussed in this review. Other severe reactions, such as anaphylaxis and vasculitis, are discussed elsewhere in this issue. Although most of the readers, particularly those in the outpatient arena, will not be treating these patients, they are the ones who will see them first, diagnose them, realize the potential danger in their condition, and refer them to the appropriate treatment venue. Therefore, dermatologists should be familiar with these conditions and be prepared to handle them adequately. PMID- 15802212 TI - The life-threatening complications of dermatologic therapies. AB - Dermatologists use a variety of systemic drugs, some of which can cause severe adverse reactions and even fatalities. Ivermectin, a well-tolerated drug, can cause severe neurological side effects, whereas metronidazole, in high cumulative doses, has been associated with convulsions and rarely with hepatotoxicity. Dapsone is associated with frequent hematologic side effects, such as methemoglobinemia, hemolysis, and anemia. Although hepatotoxicity is rare and usually mild and reversible with the new antifungal agents, severe cutaneous reactions (such as toxic epidermal necrolysis, Stevens-Johnson syndrome, and anaphylaxis) have been reported. Even a relatively safe drug such as acyclovir has been reported to be the cause of renal failure and neurotoxicity. Retinoids can cause not so benign "benign" intracranial hypertension. Methotrexate can cause not only liver toxicity, but also myelosuppression and pancytopenia, which may be acute and life threatening. Nephrotoxicity is a well-recognized side effect of cyclosporine, whereas thrombotic thrombocytopenic purpura, which is associated with high morbidity and mortality, is less well known. Dermatologists should be familiar with these and other severe adverse reactions of the most popular and most used systemic medications of our trade. PMID- 15802213 TI - Life-threatening severe allergic reactions: urticaria, angioedema, and anaphylaxis. AB - Urticaria, angioedema and anaphylaxis are life threatening skin diseases. Allergological emergencies are common; drugs, food, food ingredients/additives, insects, and many other factors have been reported to elicit anaphylactic reactions. The severity of symptoms ranges from pruritus to generalized skin eruptions, gastrointestinal, bronchial problems to systemic anaphylaxis and cardiovascular emergencies. The pathomechanisms and treatment of urticarias, angioedema and anaphylaxis are described. In some situations emergency kit: antihistamines, steroids, betamimetics and adrenaline is needed. Familiarity with the early diagnosis and correct management should be acknowledegable for dermatologists to recognize these allergic reactions and must be prepared to administer emergency kit. PMID- 15802214 TI - Life-threatening erythroderma: diagnosing and treating the "red man". AB - Exfoliative erythroderma, or diffuse erythema and scaling of the skin, may be the morphologic presentation of a variety of cutaneous and systemic diseases. Establishing the diagnosis of the underlying disease is often difficult and, not uncommonly, erythroderma is classified as idiopathic. Several cases are presented to demonstrate the diversity of presentation of this disease. Laboratory findings are typically unhelpful in establishing the etiology of erythroderma. Clinical data combined with multiple skin biopsies over time are necessary. Systemic complications of erythroderma include infection, fluid and electrolyte imbalances, thermoregulatory disturbance, high output cardiac failure, and acute respiratory distress syndrome. The initial approach to the management of erythroderma of any etiology includes attention to nutrition, fluid and electrolyte replacement, and the institution of gentle local skin care measures. Oatmeal baths and wet dressings to weeping or crusted sites should be followed by application of bland emollients and low-potency topical corticosteroids. Systemic dermatologic therapy may be required to maintain improvement achieved with local measures or to control erythroderma refractory to local measures. The prognosis of erythroderma is dependent on the underlying etiology. PMID- 15802215 TI - Transcription regulation by bacteriophage T4 AsiA. AB - Bacteriophage T4 AsiA, a strong inhibitor of bacterial RNA polymerase, was the first antisigma protein to be discovered. Recent advances that made it possible to purify large amounts of this highly toxic protein led to an increased understanding of AsiA function and structure. In this review, we discuss how the small 10-KDa AsiA protein plays a key role in T4 development through its ability to both inhibit and activate bacterial RNA polymerase transcription. PMID- 15802216 TI - Expression and characterization of N-terminal domain of Epstein-Barr virus latent membrane protein 2A in Escherichia coli. AB - Latency of Epstein-Barr virus (EBV) is maintained by the transmembrane protein latent membrane protein (LMP) 2A, which mimics the B-cell receptor (BCR) and perturbs BCR signaling. LMP2A contains a cytoplasmic N-terminal domain composed of 119 amino acids, which provides signals that are responsible for the association with various signal molecules, resulting in negative regulation of B cell signaling and the EBV lytic cycle. In the present study, to obtain N terminal domain of LMP2A (LMP2A NTD, 13 kDa) in Escherichia coli for structural analysis, a strategy for obtaining the unfused form of LMP2A NTD without any fusion partners was proposed. Recombinant LMP2A NTD has previously been expressed using the GST fusion system in E. coli [Virology 268 (2000) 178, J. Virol. 71 (1997) 4752, Mol. Cell. Biol. 20 (2000) 8526]. However, we were unable to obtain untagged LMP2A NTD from this construct because of rapid proteolysis by thrombin. To overcome the proteolysis by thrombin, C-terminal His-tagged LMP2A NTD and intein-fused LMP2A NTD were prepared. As a result, LMP2A NTD without a fusion partner could be successfully obtained using non-enzymatic cleavage. The secondary structure of the recombinant LMP2A NTD was analyzed using circular dichroism. In aqueous solution, LMP2A NTD adopts an unordered structure, which was not affected by varying pH and salt concentration. In addition, any secondary structural components of LMP2A NTD were not induced in the membrane-mimicking environments, suggesting that LMP2A NTD may intrinsically have a random coil-like structure. The biological activity of recombinant LMP2A NTD was monitored by chemical shift perturbation in HSQC spectra of LMP2A NTD with or without WW domains, which result supports that the structural change induced by WW domains is restricted within narrow region. PMID- 15802217 TI - Overexpression of human alanine:glyoxylate aminotransferase in Escherichia coli: renaturation from guanidine-HCl and affinity for pyridoxal phosphate co-factor. AB - Alanine:glyoxylate aminotransferase-1 (AGT) is a human liver peroxisomal enzyme whose deficiency results in, primary hyperoxaluria type 1 (PH1), a fatal metabolic disease. AGT requires a pyridoxal phosphate (PLP) co-factor in its active site. The AGT gene usually exists in one of two polymorphic forms, the major and minor alleles. We describe here an overexpression system for normal and mutant variants of human AGT in Escherichia coli BL21 (DE3) pLysS. We have extracted functional AGT from inclusion bodies using guanidine-HCl. Denaturation and re-folding of the overexpressed AGT after guanidine-HCl treatment produces high yields of biologically active protein and provides a strategy for generating an apoenzyme to investigate PLP-binding. K(M)s for PLP were determined by reconstitution of the apoenzyme. Successful folding was independent of the presence of PLP. The K(M) for PLP for minor allele AGT was significantly higher than that for major allele AGT. This decreased affinity could be attributed to I340M, a polymorphism associated with the minor allele. G170R, located on the minor allele and the most common PH1 mutation, had no effect on the affinity for PLP. PH1 mutations, G41V and G41R, showed enhanced activity after re-folding. We suggest that the renaturation/re-folding and reconstitution strategies provide an approach for studying the maturation of AGT under optimal conditions and in isolation from cellular quality control and chaperoning processes. Furthermore, our data show that mutations with serious consequences in vivo may not be inherently catalytically inactive and may be rescuable. PMID- 15802218 TI - Expression of G protein coupled receptors in a cell-free translational system using detergents and thioredoxin-fusion vectors. AB - In Escherichia coli and other cell-based expression systems, there are critical difficulties in synthesizing membrane proteins, such as the low protein expression levels and the formation of insoluble aggregates. However, structure determinations by X-ray crystallography require the purification of milligram quantities of membrane proteins. In this study, we tried to solve these problems by using cell-free protein expression with an E. coli S30 extract, with G protein coupled receptors (GPCRs) as the target integral membrane proteins. In this system, the thioredoxin-fusion vector induced high protein expression levels as compared with the non-fusion and hexa-histidine-tagged proteins. Two detergents, Brij35 and digitonin, effectively solubilized the produced GPCRs, with little or no effect on the protein yields. The synthesized proteins were detected by Coomassie brilliant blue staining within 1h of reaction initiation, and were easily reconstituted within phospholipid vesicles. Surprisingly, the unpurified, reconstituted thioredoxin-fused receptor proteins had functional activity, in that a specific affinity binding value of an antagonist was obtained for the receptor. This cell-free translation system (about 1mg/ml of reaction volume for 6-8 h) has biophysical and biochemical advantages for the synthesis of integral membrane proteins. PMID- 15802219 TI - Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3. AB - A thermophilic isolate Bacillus coagulans BTS-3 produced an extracellular alkaline lipase, the production of which was substantially enhanced when the type of carbon source, nitrogen source, and the initial pH of culture medium were consecutively optimized. Lipase activity 1.16 U/ml of culture medium was obtained in 48 h at 55 degrees C and pH 8.5 with refined mustard oil as carbon source and a combination of peptone and yeast extract (1:1) as nitrogen sources. The enzyme was purified 40-fold to homogeneity by ammonium sulfate precipitation and DEAE Sepharose column chromatography. Its molecular weight was 31 kDa on SDS-PAGE. The enzyme showed maximum activity at 55 degrees C and pH 8.5, and was stable between pH 8.0 and 10.5 and at temperatures up to 70 degrees C. The enzyme was found to be inhibited by Al3+, Co2+, Mn2+, and Zn2+ ions while K+, Fe3+, Hg2+, and Mg2+ ions enhanced the enzyme activity; Na+ ions have no effect on enzyme activity. The purified lipase showed a variable specificity/hydrolytic activity towards various 4-nitrophenyl esters. PMID- 15802220 TI - Temperature, media, and point of induction affect the N-terminal processing of interleukin-1beta. AB - The expression of recombinant proteins in bacterial hosts may alter the biophysical properties of the protein of interest as a result of differences in post-translational processing from that observed when produced in the native cell. For example, recombinant human interleukin-1beta (IL-1beta) is produced as three isoforms when expressed in the Escherichia coli strain BL-21(DE3). These isoforms are produced by the non-homogeneous processing of the N-terminal methionine residue by the endogenous bacterial aminopeptidase and differ in the first residue (1-met, 1-ala, and 1-pro). Importantly, these isoforms have significantly different binding affinities for the IL receptor protein. Varying the temperature, media composition, and point of induction affects this N terminal processing to favor one of the three isoforms of IL-1beta. We found changes in media composition and/or point of induction affected the abundance of the isoforms by as much as 15-fold. The 1-pro isoform decreased from 60.9 to 4.7% in Luria broth (LB) and minimal media (MM), respectively, when protein expression was induced at an OD600 of 0.9. Conversely, the abundance of the 1-met isoform is much higher in MM than in LB showing the reverse effect (2.6 and 50.7% in LB and MM, respectively, at an OD600 of 0.9), and the degree to which they are favored depends significantly upon the induction point. Our results show that it is possible to favor the expression of various N-terminal isoforms of IL-1beta by adjusting the environmental growth conditions. Given that the initiator methionine residue is necessary for expression in bacterial hosts and is known to affect the stability of other recombinant proteins our approach may be a useful general method for determining the optimal conditions for expressing and purifying pure, homogenous samples of recombinant proteins for structural and biological studies. PMID- 15802221 TI - Refolding and purification of non-fusion HPT protein expressed in Escherichia coli as inclusion bodies. AB - The gene encoding hygromycin B phosphotransferase (hpt) is a widely used selectable marker in the production of genetically engineered crops. To facilitate the safety assessment of this protein, the non-fusion hpt expression plasmid was constructed and introduced into Escherichia coli to produce enough quantity of the HPT protein. High level expressed HPT was achieved but most of the expressed protein aggregated as inclusion bodies. The inclusion bodies were washed, separated from the cells, and solubilized by 0.3% Sarkosyl. The protein was renatured by dilution and dialysis, and then purified by anion-exchange chromatography. The activity is 8 U/mg protein and the purity is about 95%. Further studies showed that the microbially produced HPT protein had comparable molecular weight, immuno-reactivities, N-terminal amino acid sequences, and biological activities with those of the HPT produced by transgenic rice harboring hpt gene. All these results demonstrated the validity of utilizing the microbially produced HPT to assess the safety of the HPT protein produced in genetically engineered rice. PMID- 15802222 TI - An approach to prevent aggregation during the purification and crystallization of wild type acyl coenzyme A: isopenicillin N acyltransferase from Penicillium chrysogenum. AB - Acyl coenzyme A: isopenicillin N acyltransferase (AT) from Penicillium chrysogenum is an enzyme of interest for the biosynthesis of beta-lactam antibiotics. Severe aggregation problems with wild type AT have, however, prevented significant progress in the structure-function analysis of this enzyme for a decade. In this study, we show an approach to solve this aggregation problem by using dynamic light scattering (DLS) analysis to probe the aggregation state of the protein in the presence of various additives. After a one-step purification of recombinant wild type AT with a C-terminal His-tag using Ni2+ affinity chelate chromatography, addition of a combination of 5 mM DTT, 250 mM NaCl, and 5 mM EDTA to the purified AT effectively prevented aggregation. In the presence of these additives, the DLS profile of AT shows a narrow size distribution indicative of a homogeneous protein solution and the absence of aggregation. The purity and mono-dispersity of wild type AT was sufficient for the growth of high quality crystals diffracting to 1.64 A resolution. PMID- 15802223 TI - Purification and characterization of a single-chain chimeric anti-p185 antibody expressed by CHO-GS system. AB - Monoclonal antibody A21 reacts specifically with the extracellular domain of p185c-erbB-2 oncoprotein, a member of the epidermal growth factor receptor family. In a previous study, we constructed a single-chain chimeric antibody, assembled using an A21 single-chain Fv antibody and a human IgG1 Fc fragment. In this study, we expressed this chimeric antibody using a CHO-GS system, and developed a simple and efficient method for its purification. After only one step using affinity purification, the recovery rate and purity of the antibody attained was 60 and 91%, respectively. After a second step, using reverse phase HPLC purification, the purity was above 99%. The high purity of the recombinant antibody allowed us to identify a number of its intrinsic molecular properties, including antigen binding activity, measurement of affinity constant, N-terminal sequencing, and mass spectrometer analysis. These results further augment the potential of this recombinant antibody to be a drug candidate for cancer therapy. PMID- 15802224 TI - High-level expression of Myrothecium verrucaria bilirubin oxidase in Pichia pastoris, and its facile purification and characterization. AB - Bilirubin oxidase (BO) from Myrothecium verrucaria (authentic BO) catalyzing the oxidation of bilirubin to biliverdine was overexpressed in the methylotrophic yeast, Pichia pastoris. The cDNA encoding BO was cloned into the P. pastoris expression vector pPIC9K under the control of the alcohol oxidase 1 promoter and its protein product was secreted using the Saccharomyces cerevisiae alpha-mating factor signal sequence. The productivity of recombinant BO (rBO) in P. pastoris was approximately 5000 U/L of culture broth, being about 2.5- and 250-fold higher than rBO expressed in Aspergillus oryzae and S. cerevisiae, respectively. The calculated molecular mass of rBO consisting of 538 amino acids was 60,493 kDa, however, that of SDS-PAGE was 66 kDa because of non-native type N-linked sugar chains. The spectroscopic properties of rBO were typical of multicopper oxidase containing four Cu ions per protein molecule. The specific activity to oxidize bilirubin was 57 U/mg, having a value about twice that of authentic BO and rBO expressed in A. oryzae. Moreover, the thermostability of rBO expressed in P. pastoris was significantly high compared to the authentic BO previously reported. Accordingly, a heterologous expression system of rBO to meet clinical and industrial needs was constructed. PMID- 15802225 TI - High level expression of peptides and proteins using cytochrome b5 as a fusion host. AB - A novel fusion protein system based on the highly soluble heme-binding domain of cytochrome b5 has been designed. The ability of cytochrome b5 to increase the levels of expression and solubility of target proteins has been tested by expressing several proteins and peptides, viz., alpha hemoglobin stabilizing protein, the regulatory subunits of acetohydroxy acid synthase I (ilvM) and II (ilvN), the carboxy terminal domains of mouse neuronal kinesin and pantothenate synthatase, two peptide toxins from cone snails, and the inactivation gate from the brain voltage gated sodium channel, NaV1.2. The fusion protein system has been designed to incorporate protease cleavage sites for commonly used proteases, viz., enterokinase, Factor Xa, and Tobacco etch virus protease. Accumulation of expressed protein as a function of time may be visually ascertained by the fact that the cells take on a bright red color during the course of induction. In all the cases tested so far, the fusion protein accumulates in the soluble fraction to high levels. A novel purification protocol has been designed to purify the fusion proteins using metal affinity chromatography, without the need of a hexahistidine-tag. Mass spectral analysis has shown that the fusion proteins are of full length. CD studies have shown that the solubilized fusion proteins are structured. The proteins of interest may be cleaved from the parent protein by either chemical or enzymatic means. The results presented here demonstrate the versatility of the cytochrome b5 based fusion system for the production of peptides and small proteins (<15 kDa). PMID- 15802227 TI - The Bordetella pertussis virulence factor P.69 pertactin retains its immunological properties after overproduction in Escherichia coli. AB - Bordetella pertussis is re-emerging in several countries with a high vaccine uptake. Analysis of clinical isolates revealed antigenic divergence between vaccine strains and circulating strains with respect to P.69 pertactin. Polymorphisms in P.69 pertactin are mainly limited to regions comprised of amino acid repeats, designated region 1 and region 2. Region 1 flanks the RGD motif involved in adherence. Although antibodies against P.69 pertactin are implicated in protective immunity, little is known about the structure and location of its epitopes. Previously we described the localization of mainly linear epitopes of both human sera and mouse monoclonal antibodies (mAbs). To study the location of conformational epitopes and to investigate the effect of variation in P.69 pertactin on vaccine efficacy, we cloned, expressed, and purified 3 naturally occurring P.69 pertactin variants, 3 mutants in which the variable regions are missing, 3 N-terminal mutants and 1 C-terminal deletion mutant. Here, we describe the procedure to clone, express, and purify up to 0.1mg P.69 pertactin and its derivatives per 1 ml Escherichia coli culture. PMID- 15802226 TI - Comparison of affinity tags for protein purification. AB - Affinity tags are highly efficient tools for purifying proteins from crude extracts. To facilitate the selection of affinity tags for purification projects, we have compared the efficiency of eight elutable affinity tags to purify proteins from Escherichia coli, yeast, Drosophila, and HeLa extracts. Our results show that the HIS, CBP, CYD (covalent yet dissociable NorpD peptide), Strep II, FLAG, HPC (heavy chain of protein C) peptide tags, and the GST and MBP protein fusion tag systems differ substantially in purity, yield, and cost. We find that the HIS tag provides good yields of tagged protein from inexpensive, high capacity resins but with only moderate purity from E. coli extracts and relatively poor purification from yeast, Drosophila, and HeLa extracts. The CBP tag produced moderate purity protein from E. coli, yeast, and Drosophila extracts, but better purity from HeLa extracts. Epitope-based tags such as FLAG and HPC produced the highest purity protein for all extracts but require expensive, low capacity resin. Our results suggest that the Strep II tag may provide an acceptable compromise of excellent purification with good yields at a moderate cost. PMID- 15802228 TI - Expression, purification, and characterization of rat protein tyrosine phosphatase eta catalytic domain. AB - Receptor-like protein tyrosine phosphatases generally contain one or two conserved intracellular catalytic domains with a conserved sequence motif ([I/V]HCXAGXXR[S/T]G), a single transmembrane domain, and an external highly variable part. Here, we describe cloning of the intracellular catalytic domain of the rat protein tyrosine phosphatase eta (rPTPetaCD) into pET28a(+) vector, its expression in Escherichia coli, purification and initial characterization. The purification of His6-tagged rPTPetaCD to near homogeneity was achieved by a combination of affinity and size exclusion chromatography. The His-tag was subsequently removed by thrombin digestion. PhastGel IEF electrophoresis demonstrated that the isoelectric point of this 41 kDa His6-tag free recombinant protein was 7.3, which is just slightly higher than the theoretically predicted value of 7.2. To assess the functionality of the rPTPetaCD we used the pNPP hydrolysis assay and observed that the enzyme has a specific activity of 9 nmol/min/mug. The secondary structure and stability of the recombinant protein was also analyzed by circular dichroism and fluorescence spectroscopy. In summary, the rPTPetaCD is stable at 18 degrees C, properly folded, and fully active, which makes it a suitable candidate for structural and functional studies. PMID- 15802229 TI - Rapid purification of EGFP, EYFP, and ECFP with high yield and purity. AB - Most current high throughput purification procedures for the green fluorescent protein (GFP) suffer from poor yields and low purity. An improved purification procedure that delivers highly pure protein (>95% homogeneity) in high yields (>70% of the initial fluorescent protein content) has been developed. The purification procedure requires only two steps: the cell lysate is heated to 60 degrees C for 4 min in ammonium sulfate and triethylamine, followed by hydrophobic interaction chromatography using isopropanol during the elution phase. The resulting pure product exhibits the same fluorescence profile as the crude sample. This procedure has been demonstrated on three commercial variants of GFP from Aequorea victoria, enhanced green, enhanced yellow, and enhanced cyan fluorescent protein (Becton-Dickinson). The yield and purity of material are superior to other recently described methods. PMID- 15802230 TI - Efficient bacterial expression of recombinant potato mop-top virus non-structural triple gene block protein 1 modified by progressive deletion of its N-terminus. AB - To obtain strong bacterial expression of proteins that seem to be hard to express in bacteria or are highly toxic for bacteria, it is possible to create a palette of similar constructs, differing only by several nucleotides, gradually deleted from the full-length clone by exonuclease III. When a construct is equipped with the 6xHis tag, a simple colony-blot procedure can be performed and a colony giving strong and efficient expression can easily be selected for high range protein expression. We utilized this procedure to produce one of potato mop-top virus (PMTV) movement proteins, namely triple gene block protein 1 (TGBp1) which was very hard to express in bacteria in its original length. The TGBp1 gene was digested with exonuclease III and nuclease S1 from its 5' terminus, leaving 6xHis tag intact. The clone that showed the strongest signal with anti-His antibodies in colony-blot procedure was found to have 44 amino acids (of total 463) deleted. The SDS-PAGE and Western blot of high range bacterial culture lysate confirmed the efficient expression of this deleted 6xHis tagged TGBp1 fragment. PMID- 15802231 TI - A custom-designed recombinant multiepitope protein as a dengue diagnostic reagent. AB - Currently, dengue fever is the most important re-emerging mosquito-borne viral disease, with the major proportion of the target population residing in the developing countries of the world. In endemic areas, potentially fatal secondary dengue infections, characterized by high anti-dengue IgG antibody titers, are most common. Most currently available commercial dengue diagnostic kits rely on the use of whole virus antigens and are consequently associated with false positives due to serologic cross-reactivity, high cost of antigen production, and biohazard risk. This has prompted the need to develop an alternate antigen to replace the whole virus antigen in diagnostic tests. We have designed and expressed a novel recombinant protein antigen by assembling key immunodominant linear IgG-specific dengue virus epitopes, chosen on the basis of pepscan analysis, phage display, and computer predictions. The recombinant dengue multiepitope protein was expressed to high levels in Escherichia coli, purified in a single step, yielding >25 mg pure protein per liter culture. We developed an in-house enzyme-linked immunosorbent assay (ELISA) to detect anti-dengue antibodies in a panel of 20 patient sera using the purified recombinant dengue multiepitope protein as the capture antigen. The ELISA results were in excellent agreement with those obtained using a commercially available diagnostic test, Dengue Duo rapid strip test from PanBio, Australia. The high epitope density, careful choice of epitopes, and the use of E. coli system for expression, coupled to simple purification, jointly have the potential to lead to the development of an inexpensive diagnostic test with a high degree of sensitivity and specificity. PMID- 15802232 TI - Malonate-assisted purification of human caspases. AB - Caspases have in the past decade become some of the most intensely pursued targets for the design of small-molecule inhibitors. Two significant technological roadblocks to developing caspase-binding molecules are the poor solubility of a subset of the bacterially expressed proteins and the instability of the renatured proteins that results from rapid inactivating autolysis at high protein concentrations. In this report, we present a generalized method of renaturing human caspases and inhibiting the self-proteolytic activity of the enzymes without a need for covalent active-site inhibitors. Our method, which involves blocking the S1 region of the active site with malonate, enables one to inhibit fully the inactivating autolysis in human caspases and increases the yields of renatured active enzyme. It furthermore does not necessitate removal of malonate prior to setting up enzymatic assays since as high as 100-mM concentrations of malonate do not compete efficiently with caspase substrates or larger caspase inhibitors for binding to the active site. The method described in this report simplifies greatly caspase purification and makes it possible to stabilize the enzymes against autolysis without a need for costly, and frequently synthetically challenging, small-molecule inhibitors. PMID- 15802233 TI - Purification and biochemical characterization of a heme containing peroxidase from the human parasite P. falciparum. AB - A peroxidase (30 kDa) has been purified from the human malaria parasite Plasmodium falciparum to its homogeneity. The protein is a dimer of 15 kDa subunit as evident from SDS-PAGE and MALDI-TOF mass analysis. The antibodies developed against the purified protein cross-react selectively with this protein present in parasite lysate. It is a heme containing peroxidase [R/Z value (A408/A278)=2.33] showing characteristic heme spectra with Soret peak at 408 nm and visible peaks at 536 and 572 nm. Analysis of Soret spectra in presence or absence of cyanide or azide reveals that iron of heme is in Fe-III state. Circular dichroism spectral analysis establishes that this protein contains mainly alpha-helix (60-70%). H2O2 interacts with the heme moiety of the enzyme as evidenced by optical difference spectroscopy and spectral studies indicate the formation of catalytically active peroxidase-H2O2 complex (Soret peak at 413 nm) to exhibit peroxidase activity. During the erythrocytic stages of its life cycle, the parasite is exposed to oxidative stress. As the parasite is susceptible to oxidative stress, this peroxidase may offer antioxidant role by scavenging endogenous H2O2. PMID- 15802234 TI - Expression and purification of active PKB kinase from Escherichia coli. AB - PKB/Akt is a protein involved in control of apoptosis, proliferation and cellular metabolism, and it has been found to be activated in many cancers. Activation of PKB involves recruitment of the enzyme by its PH domain to the cell membrane, and phosphorylation at two residues, T308 and S473. To produce active PKB kinase from Escherichia coli, we constructed a derivative of PKB lacking the PH domain and mutated to glutamate at residues S124, T450 and the activating residue S473 (DeltaPH-PKB-EEE). DeltaPH-PKB-EEE was expressed in E. coli together with PDK1, the kinase responsible for phosphorylating PKB at T308, which was expressed as a GST-fusion. Full-length DeltaPH-PKB-EEE was obtained by using a double tag strategy: His6 at the N-terminus and FLAG at the C-terminus. The protein was purified by nickel affinity chromatography, followed by passage over an anti-FLAG column. The final purification step, anion exchange over a monoQ column, separated phosphorylated from unphosphorylated protein. Active recombinant PKB kinase was thus produced from E. coli, by a simple, reproducible procedure. PMID- 15802235 TI - Prokaryotic expression and polyclonal antibody preparation of novel ZLG10 protein involved in infection of RSV on SPC-A1 cells. AB - Differentially expressed genes between normal SPC-A1 cells and SPC-A1 cells infected by RSV were investigated using differential display. The novel zlg10 gene codes for a novel protein, ZLG10, which has previously been reported to be up-regulated in RSV-infected SPC-A1 cells. Its putative open reading frame was also identified. To better understand the structure, function, and possible role of ZLG10 as a potential candidate for diagnosis and vaccine studies, the intact region encoding ZLG10 was obtained by PCR and expressed in Escherichia coli as a GST-fusion protein. After purification, GST-ZLG10 fusion protein was used to immunize the adult rabbits following standard protocols. Consequently, we found that the produced antiserum of the novel fusion protein significantly suppressed the infection by RSV on SPC-A1 cells by using neutral red uptake assay and quantitative measurement. Together, our data demonstrate that ZLG10, a novel protein expressed and purified in this report, might be a potential effective therapeutic candidate for treating RSV infections. PMID- 15802236 TI - Expression, purification, and inhibition of human RET tyrosine kinase. AB - Tyrosine kinases are emerging as frequent targets of primary oncogenic events and therefore represent an optimal focus of therapeutical intervention. Genetic alterations that cause dysregulated activation of the RET tyrosine kinase are responsible for a significant fraction of thyroid carcinomas. In an effort towards therapeutic RET inactivation, we have developed a method for expression and purification of recombinant RET catalytic domain for structural purposes and for use in the screening of potential inhibitors of RET kinase activity. His tagged RET kinase domain was purified from Sf9 insect cell lysate using a two step chromatographic protocol and characterised. Purified recombinant RET phosphorylated itself and exogenous substrates at physiological pH. A specific peptide substrate, derived from RET activation loop, was identified and experimentally validated. These reagents were used to develop a rapid ELISA-based kinase assay for screening potential inhibitors. Novel RET inhibitors were identified using this assay. PMID- 15802237 TI - Refolding, purification, and crystallization of apical membrane antigen 1 from Plasmodium falciparum. AB - Extracellular domains of malaria antigens almost invariably contain disulphide linkages but lack N- and O-linked glycosylation. The best practical approach to generating recombinant extracellular Plasmodium proteins is not established and the problems encountered when using a bacterial expression/refolding approach are discussed in detail. Limited proteolysis experiments were used to identify a relatively non-flexible core region of the Plasmodium falciparum protein apical membrane antigen 1 (AMA1), and refolding/purification was used to generate two fragments of AMA1. Several chromatographically distinct AMA1 variants were identified that are presumably differentially refolded proteins. One of these AMA1 preparations proved to be crystallizable and generated two crystal forms that diffracted X-rays to 2 A resolution. PMID- 15802238 TI - Expression, refolding, and characterization of human soluble BAFF synthesized in Escherichia coli. AB - The B lymphocyte stimulator (BAFF) is a novel member of the tumor necrosis factor (TNF) ligand family which is important in B lymphocyte maturation and survival. Here, a recombinant form of the extracellular domain of the BAFF (hsBAFF) was expressed in Escherichia coli BL21(DE3) under the control of a T7 promoter. The resulting insoluble bodies were separated from cellular debris by centrifugation and solubilized with 8 M urea. A rapid and simple on-column refolding procedure was developed. It was applied and then the refolded hsBAFF was purified by anion exchange. The purified final product was >98% pure by SDS-PAGE stained with Coomassie brilliant blue R-250. Mass spectroscopic analysis indicated the protein to be 17.5 kDa, which equalled the theoretically expected mass. The N-terminal sequencing of refolding hsBAFF showed the sequence corresponded to the designed protein. The correct refolding of the recombinant protein was verified in the recovery of its secondary and tertiary structures as assessed by circular dichroism and fluorescence emission spectra. The renatured protein displayed its immunoreactivity with the antibodies to BAFF protein by Western blotting. The final purified material was biologically active in a validated induced human B lymphocyte proliferation bioassay. The expression and in vitro refolding of hsBAFF resulted in production of an active molecule in a yield of 15 mg/L flask cultivation. PMID- 15802239 TI - Getting the signals across: networking inside and out. PMID- 15802240 TI - Collaborative signaling by bacterial chemoreceptors. AB - Motile bacteria seek optimal living habitats by following gradients of attractant and repellent chemicals in their environment. The signaling machinery for these chemotactic behaviors, although assembled from just a few protein components, has extraordinary information-processing capabilities. Escherichia coli, the best studied model, employs a networked cluster of transmembrane receptors to detect minute chemical stimuli, to integrate multiple and conflicting inputs, and to generate an amplified output signal that controls the cell's flagellar motors. Signal gain arises through cooperative action of chemoreceptors of different types. The signaling-teams within a receptor cluster may be built from trimers of receptor dimers that communicate through shared connections to their partner signaling proteins. PMID- 15802241 TI - Sensing external stress: watchdogs of the Escherichia coli cell envelope. AB - The Cpx and sigmaE signaling systems monitor the cell envelope in Escherichia coli. When induced, each system triggers a signaling cascade that leads to the upregulation of factors needed to combat envelope damage. Although each system is distinct and can be uniquely induced by certain cues, they also share striking similarities. In this review, we discuss the recent progress in our understanding of the Cpx and sigmaE systems and compare how both function to maintain the integrity of the cell envelope. PMID- 15802242 TI - The bgl sensory system: a transmembrane signaling pathway controlling transcriptional antitermination. AB - The bgl system represents a family of sensory systems composed of membrane-bound sugar-sensors and transcriptional antiterminators, which regulate expression of genes involved in sugar utilization in response to the presence of the corresponding sugar in the growth medium. The BglF sensor catalyzes different activities depending on its stimulation state: in its non-stimulated state, it phosphorylates the BglG transcriptional regulator, thus inactivating it; in the presence of the stimulating sugar, it transports the sugar and phosphorylates it and also activates BglG by dephosphorylation, leading to bgl operon expression. The sugar stimulates BglF by inducing a change in its membrane topology. BglG exists in several conformations: a dimer, which is active, and compact and non compact monomers, which are inactive. BglF modulates the transition of BglG from one conformation to another, depending on sugar availability. The two Bgl proteins form a pre-complex at the membrane that dissociates upon stimulation, enabling BglG to exert its effect on transcription. PMID- 15802243 TI - Role of membrane lipids in bacterial division-site selection. AB - Heterogeneous distribution of specific phospholipids along the bacterial membrane results in the formation of domains enriched in anionic phospholipids at the cell poles and cell center, which appear to participate in the binding of amphitropic proteins responsible for selection and recognition of the division site. It was discovered that functioning of the Min system, which protects the cell poles from aberrant positioning of the Z-ring, is controlled by direct interaction of its MinD component with membrane phospholipids. There is also an accumulation of evidence that the mid-cell domain, formed in the cell at a defined step of the cell cycle, provides the optimal phospholipid composition first for initiation of DNA replication and then for Z-ring positioning. PMID- 15802244 TI - Chromosomal replication and the cell membrane. AB - The importance of the cell membrane in bacterial chromosomal replication continues to emerge. Recent advances include better definition of the biochemical interaction between membrane acidic phospholipids and the replication initiator, DnaA protein, the physiological impact that an altered membrane lipid composition has on chromosomal replication and the identification and characterization of recently identified membrane-associated proteins that regulate replication and participate in chromosomal segregation. PMID- 15802245 TI - Transcriptional control of membrane lipid synthesis in bacteria. AB - Fatty acid synthesis is coordinately regulated with phospholipid, macromolecular synthesis and growth as part of the response to changes in the environment. Many of these processes are rapid responses of the integrated biochemical network and do not involve changes in gene expression. An important recent development is the identification and characterization of transcription factors that modify pathway activity by either altering the expression levels of a few important genes or controlling a global adjustment in the expression of the entire pathway. For most of these transcription factors the signaling molecules controlling their activities are still poorly defined. PMID- 15802246 TI - Dam methylation: coordinating cellular processes. AB - GATC sequences in Escherichia coli DNA are methylated at the adenine residue by DNA adenine methyltransferase (DamMT). These methylated residues and/or the level of DamMT can influence cellular functions such as gene transcription, DNA mismatch repair, initiation of chromosome replication and nucleoid structure. In certain bacteria, unlike E. coli, DamMT is essential for viability perhaps owing to its role in chromosome replication. DamMT has also been implicated as a virulence factor in bacterial pathogenesis. The origin and phylogeny of DamMT, based on sequenced genomes, has been deduced. PMID- 15802248 TI - PII signal transduction proteins: sensors of alpha-ketoglutarate that regulate nitrogen metabolism. AB - PII proteins are small homotrimeric signal transduction proteins that regulate the activities of metabolic enzymes and permeases, and control the activities of signal transduction enzymes. The protein family shows high conservation, with examples in eukaryota (plants and eukaryotic algae), archaea, and bacteria. This distribution indicates that PII is one of the most ancient signalling proteins known. PMID- 15802247 TI - Structural and mechanistic diversity of secondary transporters. AB - Recent reports on the three-dimensional structure of secondary transporters have dramatically increased our knowledge of the translocation mechanism of ions and solutes. The structures of five transporters at atomic resolution have yielded four different folds and as many different translocation mechanisms. The structure of the glutamate transporter homologue GltPh confirmed the role of pore loop structures as essential parts of the translocation mechanism in one family of secondary transporters. Biochemical evidence for pore-loop structures in several other families suggest that they might be common in secondary transporters, adding to the structural and mechanistic diversity of secondary transporters. PMID- 15802249 TI - Protein targeting by the bacterial twin-arginine translocation (Tat) pathway. AB - The Tat (twin-arginine translocation) protein export system is found in the cytoplasmic membrane of most prokaryotes and is dedicated to the transport of folded proteins. The Tat system is now known to be essential for many bacterial processes including energy metabolism, cell wall biosynthesis, the nitrogen fixing symbiosis and bacterial pathogenesis. Recent studies demonstrate that substrate-specific accessory proteins prevent improperly assembled substrates from interacting with the Tat transporter. During the transport cycle itself substrate proteins bind to a receptor complex in the membrane which then recruits a protein-translocating channel to carry out the transport reaction. PMID- 15802250 TI - YidC--an evolutionary conserved device for the assembly of energy-transducing membrane protein complexes. AB - Members of the YidC/Oxa1/Alb3 membrane protein family are multifunctional mediators of membrane protein integration, folding and assembly into large complexes. Their evolutionary conserved and physiologically important role appears to relate to the assembly of major energy-transducing membrane protein complexes. PMID- 15802251 TI - Metal ion homeostasis in Bacillus subtilis. AB - Bacillus subtilis, a Gram-positive soil bacterium, provides a model system for the study of metal ion homeostasis. Metalloregulatory proteins serve as the arbiters of metal ion sufficiency and regulate the expression of metal homeostasis pathways. In B. subtilis, uptake systems are regulated by the highly selective metal-sensing repressors Fur (iron), Zur (zinc), and MntR (manganese). Metal efflux systems are regulated by MerR and ArsR family homologs which, by contrast, can be rather non-specific with regard to metal selectivity. A Fur homolog, PerR, functions as an Fe(II)-dependent peroxide stress sensor and regulates putative metal transport and storage functions. PMID- 15802252 TI - Bacterial zinc uptake and regulators. AB - Many bacteria use an ABC transporter for high-affinity uptake of zinc with a cluster 9 solute-binding protein. Other members of this protein family transport manganese. At present, it is not always possible to distinguish zinc-specific and manganese-specific transporters on the basis of sequence analysis. Low-affinity ZIP-type zinc transporters in bacteria have also been identified. Most high affinity zinc uptake systems are regulated by Zur proteins, which form at least three unrelated subgroups of the Fur protein family (regulators of iron transport). High-affinity transport of zinc out of the periplasmic space poses a problem to the cell because zinc is a cofactor of several periplasmic enzymes. Certain zinc-binding proteins in the periplasm might function as chaperones to supply these enzymes with zinc. PMID- 15802253 TI - CodY, a global regulator of stationary phase and virulence in Gram-positive bacteria. AB - Many Gram-positive bacteria encode a homolog of Bacillus subtilis CodY, a protein that controls more than a hundred genes that are typically repressed during rapid growth and induced when cells experience nutrient deprivation. In B. subtilis, the repressor function of CodY is activated by interaction with two different effectors, GTP and isoleucine, which independently and additively increase the affinity of CodY for its target sites. In at least some pathogenic Gram-positive bacteria, major virulence factor genes are among the targets of CodY. PMID- 15802254 TI - Regulation of secondary metabolism in streptomycetes. AB - While the biological functions of most of the secondary metabolites made by streptomycetes are not known, it is inconceivable that they do not play an adaptive ecological role. The biosynthesis of secondary metabolites under laboratory conditions usually occurs in a growth phase or developmentally controlled manner, but is also influenced by a wide variety of environmental and physiological signals, presumably reflecting the range of conditions that trigger their production in nature. The expression of secondary metabolic gene clusters is controlled by many different families of regulatory proteins, some of which are found only in actinomycetes, and is elicited by both extracellular and intracellular signalling molecules. The application of a variety of genetic and molecular approaches is now beginning to reveal fascinating insights into the complex regulatory cascades that govern this process. PMID- 15802255 TI - Reversing cell polarity: evidence and hypothesis. AB - The long, rod-shaped cells of myxobacteria are polarized by their gliding engines. At the rear, A-engines push while pili pull the front end forward. An hypothesis is developed whereby both engines are partially dis-assembled, then re assembled at the opposite pole when cells reverse their movement direction. Reversals are induced by an Mgl G-protein switch that controls engine polarity. The switch is driven by an oscillatory circuit of Frizzy proteins. In growing cells, the circuit gives rise to an occasional reversal that makes swarming possible. Then, as myxobacteria begin fruiting body development, a rising level of C-signal input drives the oscillator and changes the reversal pattern. Cells reverse regularly every eight minutes in traveling waves, the reversal period is then prolonged enabling cells to form streams that enlarge tiny random aggregates into fruiting bodies. PMID- 15802256 TI - Lessons from DNA microarray analysis: the gene expression profile of biofilms. AB - DNA microarray technology has been used to identify the global gene expression profile of biofilm cells. This is an interesting case study in how DNA microarray technology has advanced the molecular understanding of an understudied research area. DNA microarray analyses have suggested that there may be common responses upon biofilm formation, such as the repression of flagella genes and hyper expression of genes for adhesion and ribosomal protein. They have also assisted in the identification of transcription factors that affect the formation of biofilms and indicated that there may not be biofilm-specific genes, arguing against biofilm formation being a developmental process. Instead, the DNA microarray data suggest that biofilms may have a unique pattern of gene expression, in which sub-sets of genes expressed in biofilms are also expressed under different planktonic conditions, but only in the biofilm are they all expressed simultaneously. PMID- 15802257 TI - Comparing night-to-night variability of sleep measures in elderly African Americans and Whites. AB - There has been great interest in the night-to-night variability of sleep disordered breathing because of the implications for the accuracy and costs associated with diagnostic testing. Night-to-night variability of sleep and respiratory parameters were examined in 66 older Whites and 56 older African Americans. Each participant had his or her sleep recorded for 2 nights with portable recording equipment. The correlations between the 2 nights were all large. Analyses of variance showed that men had a greater change in apnea index and in number of apneas than women. There were no differences in the parameters for African Americans versus Whites. One night of sleep recording may be sufficient for studying sleep disorders in elderly persons. PMID- 15802258 TI - Focusing on the experience of insomnia. AB - Although insomnia is defined by sleep disturbances, patients also complain of daytime problems. To better define the range of experiences associated with insomnia, we conducted 3 focus groups comprising 16 patients with chronic insomnia. Participants were asked to describe their experience of insomnia and its effects, the changes that occurred with improved sleep, and what clinicians should ask about insomnia. Participants affirmed daytime consequences that are frequently cited in the literature: fatigue, irritability, and decreased performance. In addition, members of each group reported that they felt that the impact that insomnia had on their lives was pervasive and misunderstood by others who were significant to them or treating their sleep complaints. Daytime problems are salient descriptors of the insomnia experience. Assessing the patient's daytime experiences is valued by patients with insomnia and may be used as a benchmark for treatment response. PMID- 15802259 TI - Predictors of interest in psychological treatment for insomnia among older primary care patients with disturbed sleep. AB - In this study, we examined whether the common sense model of illness representation (CSMIR) could be successfully used to predict interest in cognitive-behavioral treatment for insomnia (CBT-I) among older primary care patients with disturbed sleep. The Sleep Impairment Index (C. M. Morin, 1993) was used to assess sleep disturbance and the constructs of the CSMIR in primary care patients ages 55 and older. Statistical analyses showed that the CSMIR constructs of consequences (perceived adverse consequences of sleep disturbance to functioning), causes (attributing one's insomnia to bad sleeping habits), and emotion (concern about one's sleep problem) predicted interest in CBT-I. These data provided encouraging support for the ability of the CSMIR to accurately predict patient interest in treatment for insomnia. Implications for assessment and treatment of insomnia in primary care are discussed. PMID- 15802260 TI - Insomnia and complicated grief symptoms in bereaved college students. AB - In this study, we extended previous research by concentrating on sleep- and grief related symptoms in a cohort of bereaved college students, in view of the potential for each of these problems to exacerbate the other. A sample of 815 college students completed the Inventory of Complicated Grief (H. G. Prigerson & S. C. Jacobs, 2001), along with an assessment of diagnostic criteria for insomnia and associated sleep behaviors. As predicted, the rate of insomnia was significantly higher (22%) in the bereaved sample than in a nonbereaved comparison group (17%), a difference that was particularly pronounced in terms of middle insomnia. Also as hypothesized, bereaved insomniacs reported higher complicated grief scores than bereaved noninsomniacs, and several specific sleep variables (including sleep-onset insomnia related to nighttime rumination about the loss and sleep-maintenance insomnia associated with dreaming of the deceased) were significantly related to complicated grief symptomatology. PMID- 15802261 TI - The CentriMag: a new optimized centrifugal blood pump with levitating impeller. AB - PURPOSE: Blood pumps are routinely used for circulatory and pulmonary support. However, blood trauma and pump failure remain severe drawbacks of currently available pump models. This study evaluated the first clinical application of a new, totally bearingless centrifugal blood pump (CentriMag). MATERIAL AND METHODS: A centrifugal pump consisting of an electromagnetic suspended impeller was used as a blood pump during beating-heart coronary artery bypass grafting in 11 patients (mean weight, 77.4 kg). Heparin in a bolus of 150 IU/kg body weight was administered, and activated clotting time was maintained at approximately 180 to 250 seconds during extracorporeal circulation. Pump-induced blood trauma was evaluated by measurement of plasma free hemoglobin (PFH), lactate dehydrogenase (LDH), hematocrit, total bilirubin, and platelet levels. RESULTS: Mean pump flow was 3.3 +/- 0.62 L/min, and mean pressure gradient through the oxygenator was 69 +/- 4 mm Hg. No pump dysfunction occurred during a mean application time of 105 +/- 26 minutes. Inspection of the pump housings showed no internal thrombus formation despite low-dose heparinization. Only slight hemolysis was observed with a mean PFH level of 1.96 micromol/L; LDH, 460 U/L; hematocrit, 33%; total bilirubin, 25 micromol/L; and platelets, 191 x 10(3)/microL. CONCLUSIONS: The bearingless CentriMag blood pump is a safe and reliable new device that produces only minimal hemolysis. It seems to be suited for long-term evaluation as a blood pump for extracorporeal membrane oxygenation or as ventricular assist device. PMID- 15802262 TI - Acute insult of ammonia leads to calcium-dependent glutamate release from cultured astrocytes, an effect of pH. AB - Hyperammonemia is a key factor in the pathogenesis of hepatic encephalopathy (HE) as well as other metabolic encephalopathies, such as those associated with inherited disorders of urea cycle enzymes and in Reye's syndrome. Acute HE results in increased brain ammonia (up to 5 mM), astrocytic swelling, and altered glutamatergic function. In the present study, using fluorescence imaging techniques, acute exposure (10 min) of ammonia (NH4+/NH3) to cultured astrocytes resulted in a concentration-dependent, transient increase in [Ca2+]i. This calcium transient was due to release from intracellular calcium stores, since the response was thapsigargin-sensitive and was still observed in calcium-free buffer. Using an enzyme-linked fluorescence assay, glutamate release was measured indirectly via the production of NADH (a naturally fluorescent product when excited with UV light). NH4+/NH3 (5 mM) stimulated a calcium-dependent glutamate release from cultured astrocytes, which was inhibited after preincubation with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester but unaffected after preincubation with glutamate transport inhibitors dihydrokainate and DL-threo-beta-benzyloxyaspartate. NH4+/NH3 (5 mM) also induced a transient intracellular alkaline shift. To investigate whether the effects of NH4+/NH3 were mediated by an increase in pH(i), we applied trimethylamine (TMA+/TMA) as another weak base. TMA+/TMA (5 mM) induced a similar transient increase in both pH(i) and [Ca2+]i (mobilization from intracellular calcium stores) and resulted in calcium dependent release of glutamate. These results indicate that an acute exposure to ammonia, resulting in cytosolic alkalinization, leads to calcium-dependent glutamate release from astrocytes. A deregulation of glutamate release from astrocytes by ammonia could contribute to glutamate dysfunction consistently observed in acute HE. PMID- 15802263 TI - Peptidoglycan signaling in innate immunity and inflammatory disease. PMID- 15802264 TI - The small G-protein Arf6GTP recruits the AP-2 adaptor complex to membranes. AB - The small GTP-binding protein ADP-ribosylation factor 6 (Arf6) is involved in plasma membrane/endosomes trafficking. However, precisely how the activation of Arf6 regulates vesicular transport is still unclear. Here, we show that, in vitro, recombinant Arf6GTP recruits purified clathrin-adaptor complex AP-2 (but not AP-1) onto phospholipid liposomes in the absence of phosphoinositides. We also show that phosphoinositides and Arf6 tightly cooperate to translocate AP-2 to the membrane. In vivo, Arf6GTP (but not Arf6GDP) was found associated to AP-2. The expression of the GTP-locked mutant of Arf6 leads to the plasma membrane redistribution of AP-2 in Arf6GTP-enriched areas. Finally, we demonstrated that the expression of the GTP-locked mutant of Arf6 inhibits transferrin receptor internalization without affecting its recycling. Altogether, our results demonstrated that Arf6GTP interacts specifically with AP-2 and promotes its membrane recruitment. These findings strongly suggest that Arf6 plays a major role in clathrin-mediated endocytosis by directly controlling the assembly of the AP-2/clathrin coat. PMID- 15802265 TI - Glutamate 636 of the Escherichia coli pyruvate dehydrogenase-E1 participates in active center communication and behaves as an engineered acetolactate synthase with unusual stereoselectivity. AB - The residue Glu636 is located near the thiamine diphosphate (ThDP) binding site of the Escherichia coli pyruvate dehydrogenase complex E1 subunit (PDHc-E1), and to probe its function two variants, E636A and E636Q were created with specific activities of 2.5 and 26% compared with parental PDHc-E1. According to both fluorescence binding and kinetic assays, the E636A variant behaved according to half-of-the-sites mechanism with respect to ThDP. In contrast, with the E636Q variant a K(d,ThDP) = 4.34 microM and K(m,ThDP) = 11 microM were obtained with behavior more reminiscent of the parental enzyme. The CD spectra of both variants gave evidence for formation of the 1',4'-iminopyrimidine tautomer on binding of phosphonolactylthiamine diphosphate, a stable analog of the substrate-ThDP covalent complex. Rapid formation of optically active (R)-acetolactate by both variants, but not by the parental enzyme, was observed by CD and NMR spectroscopy. The acetolactate configuration produced by the Glu636 variants is opposite that produced by the enzyme acetolactate synthase and the Asp28 substituted variants of yeast pyruvate decarboxylase, suggesting that the active centers of the two sets of enzymes exhibit different facial selectivity (re or si) vis a vis pyruvate. The tryptic peptide map (mass spectral analysis) revealed that the Glu636 substitution changed the mobility of a loop comprising amino acid residues from the ThDP binding fold. Apparently, the residue Glu636 has important functions both in active center communication and in protecting the active center from undesirable "carboligase" side reactions. PMID- 15802266 TI - Essential role of beta-catenin in postnatal bone acquisition. AB - Mutations in the Wnt co-receptor LRP5 alter bone mass in humans, but the mechanisms responsible for Wnts actions in bone are unclear. To investigate the role of the classical Wnt signaling pathway in osteogenesis, we generated mice lacking the beta-catenin or adenomatous polyposis coli (Apc) genes in osteoblasts. Loss of beta-catenin produced severe osteopenia with striking increases in osteoclasts, whereas constitutive activation of beta-catenin in the conditional Apc mutants resulted in dramatically increased bone deposition and a disappearance of osteoclasts. In vitro, osteoblasts lacking the beta-catenin gene exhibited impaired maturation and mineralization with elevated expression of the osteoclast differentiation factor, receptor activated by nuclear factor-kappaB ligand (RANKL), and diminished expression of the RANKL decoy receptor, osteoprotegerin. By contrast, Apc-deficient osteoblasts matured normally but demonstrated decreased expression of RANKL and increased osteoprotegerin. These findings suggest that Wnt/beta-catenin signaling in osteoblasts coordinates postnatal bone acquisition by controlling the differentiation and activity of both osteoblasts and osteoclasts. PMID- 15802267 TI - Evidence for MR1 antigen presentation to mucosal-associated invariant T cells. AB - The novel class Ib molecule MR1 is highly conserved in mammals, particularly in its alpha1/alpha2 domains. Recent studies demonstrated that MR1 expression is required for development and expansion of a small population of T cells expressing an invariant T cell receptor (TCR) alpha chain called mucosal associated invariant T (MAIT) cells. Despite these intriguing properties it has been difficult to determine whether MR1 expression and MAIT cell recognition is ligand-dependent. To address these outstanding questions, monoclonal antibodies were produced in MR1 knock-out mice immunized with recombinant MR1 protein, and a series of MR1 mutations were generated at sites previously shown to disrupt the ability of class Ia molecules to bind peptide or TCR. Here we show that 1) MR1 molecules are detected by monoclonal antibodies in either an open or folded conformation that correlates precisely with peptide-induced conformational changes in class Ia molecules, 2) only the folded MR1 conformer activated 2/2 MAIT hybridoma cells tested, 3) the pattern of MAIT cell activation by the MR1 mutants implies the MR1/TCR orientation is strikingly similar to published major histocompatibility complex/alphabetaTCR engagements, 4) all the MR1 mutations tested and found to severely reduce surface expression of folded molecules were located in the putative ligand binding groove, and 5) certain groove mutants of MR1 that are highly expressed on the cell surface disrupt MAIT cell activation. These combined data strongly support the conclusion that MR1 has an antigen presentation function. PMID- 15802268 TI - Epidermal growth factor and hypoxia-induced expression of CXC chemokine receptor 4 on non-small cell lung cancer cells is regulated by the phosphatidylinositol 3 kinase/PTEN/AKT/mammalian target of rapamycin signaling pathway and activation of hypoxia inducible factor-1alpha. AB - Non-small cell lung cancer (NSCLC) expresses a particularly aggressive metastatic phenotype, and patients with this disease have a poor prognosis. CXC chemokine receptor 4 (CXCR4) is a cell surface receptor that has been shown to mediate the metastasis of many solid tumors including lung, breast, kidney, and prostate. In addition, overexpression of the epidermal growth factor receptor (EGFR) is associated with the majority of NSCLC and has been implicated in the process of malignant transformation by promoting cell proliferation, cell survival, and motility. Here we show for the first time that activation of the EGFR by EGF increases CXCR4 expression and the migratory capacity of NSCLC cells. Furthermore, many solid tumors are associated with low oxygen tension, and when NSCLC cells were cultured with EGF under hypoxic conditions, CXCR4 expression was dramatically enhanced. A molecular analysis of these events indicated that augmented CXCR4 expression was regulated by the phosphatidylinositol 3 kinase/PTEN/AKT/mammalian target of rapamycin signal transduction pathway, activation of hypoxia inducible factor (HIF) 1alpha, and ultimately HIF-1 dependent transcription of the CXCR4 gene. Thus, a combination of low oxygen tension and overexpression of EGFR within the primary tumor of NSCLC may provide the microenvironmental signals necessary to upregulate CXCR4 expression and promote metastasis. PMID- 15802269 TI - Wnt-7a up-regulates matrix metalloproteinase-12 expression and promotes cell proliferation in corneal epithelial cells during wound healing. AB - Corneal wound repair involves the rapid coverage of a denuded area by residual epithelial cells. During wound healing, there are different cell behaviors in different regions of the epithelium: cell proliferation in the peripheral epithelium and cell migration in the central epithelium. We found that Wnt-7a was rapidly induced in the wounded cornea, promoted the proliferation of corneal epithelial cells, and enhanced wound closure. Matrix metalloproteinase-12 (MMP 12) was detected in the peripheral epithelium, where cell proliferation was enhanced, but was diminished in the migrating central epithelium. Wnt-7a induced the accumulation of beta-catenin and the activation of Rac and beta-catenin, and Rac synergistically induced the transcription of MMP-12. Blocking the function of MMP-12 delayed wound closure induced by Wnt-7a. Our results also suggest that, in addition to the beta-catenin pathway, Wnt-7a might induce a beta-catenin independent pathway. By regulating the proliferation of corneal epithelial cells, Wnt-7a and MMP-12 appear to contribute to corneal wound healing. PMID- 15802270 TI - Developmental regulation of the direct interaction between the intracellular loop of connexin 45.6 and the C terminus of major intrinsic protein (aquaporin-0). AB - The eye lens is dependent upon a network of gap junction-mediated intercellular communication to facilitate its homeostasis and development. Three gap junction forming proteins are expressed in the lens of which two are in lens fibers, namely connexin (Cx) 45.6 and 56. Major intrinsic protein (MIP), also known as aquaporin-0 (AQP0), is the most abundant membrane protein in lens fibers. However, its role in the lens is not clear. Our previous studies show that MIP(AQP0) associates with gap junction plaques formed by Cx45.6 and Cx56 during the early stages of embryonic chick lens development but not in late embryonic and adult lenses. We report here that MIP(AQP0) directly interacts with Cx45.6 but not with Cx56. We further identified the intracellular loop of Cx45.6 as the interacting domain for the MIP(AQP0) C terminus. Surface plasmon resonance experiments indicated that the C-terminal domain of MIP(AQP0) interacts with two binding sites within the intracellular loop region of Cx45.6 with a K(D(app)) of 7.5 and 10.3 microm, respectively. The K(D(app)) for the full-length loop region is 7.7 microm. The cleavage at the intracellular loop of Cx45.6 was observed during lens development, and the C terminus of MIP(AQP0) did not interact with the loop-cleaved form of Cx45.6. Thus, the dissociation between these two proteins that occurs in the mature fibers of late lens development is likely caused by this cleavage. Finally this interaction had no impact on Cx45.6 mediated intercellular communication, suggesting that the Cx45.6-MIP(AQP0) interaction plays a novel unidentified role in lens fibers. PMID- 15802271 TI - Regulation of the Dbl proto-oncogene by heat shock cognate protein 70 (Hsc70). AB - The dbl oncogene product is the defining member of a family of onco-proteins known as Dbl guanine nucleotide exchange factors (GEFs) that facilitate the activation of the small GTP-binding proteins Cdc42, Rac, and Rho. Oncogenic activation of proto-Dbl occurs through loss of the amino-terminal 497 residues, rendering the protein constitutively active. Because both onco- and proto-Dbl contain the structural elements required for GEF activity (i.e. the Dbl homology (DH) and pleckstrin homology (PH) domains), it is thought that the amino terminus of proto-Dbl somehow inhibits the biochemical activity of the protein. To better understand the molecular basis of this regulation, we set forth to identify cellular proteins that preferentially bind the proto-oncogenic form of Dbl. We identified the molecular chaperone heat shock cognate protein (Hsc70) as a binding partner that preferentially interacts with the proto-oncogenic form of Dbl. Dbl is complexed with Hsc70 in transfected cells, as well as in native mouse brain extracts. The interaction between Hsc70 and proto-Dbl is mediated by at least two regions in Dbl, the aminoterminal spectrin homology domain (residues 224-417) and the pleckstrin homology domain (residues 711-808). Overexpression of a dominant negative Hsc70 mutant leads to activation of proto-Dbl GEF activity, indicating that the chaperone negatively regulates proto-Dbl function in vivo. We propose that Hsc70 attenuates Dbl activity by maintaining an inactive conformation in which the amino terminus is "folded over" the catalytic DH-PH domain. PMID- 15802272 TI - Internal translation initiation mediated by the angiogenic factor Tie2. AB - Tie2 is an endothelium-specific receptor tyrosine kinase required for normal blood vessel maturation. We report that Tie2 mRNA translation is maintained under hypoxic conditions. To identify the mechanism responsible for this, we undertook structure/function analysis of the Tie2 5'-untranslated region (UTR). Transcription start site mapping indicates the existence of a several mRNA isoforms containing unusually long 5'-UTRs (>350 nucleotides) with five upstream open reading frames. We find internal ribosome binding activity that allows the Tie2 mRNA to initiate in a cap-independent fashion. Our data provide a framework for understanding how Tie2 mRNA is translated despite a cumbersome structured 5' UTR and how its production is secured under unfavorable environmental conditions. PMID- 15802273 TI - Caveolin-1 promotes tumor progression in an autochthonous mouse model of prostate cancer: genetic ablation of Cav-1 delays advanced prostate tumor development in tramp mice. AB - Caveolin-1 (Cav-1) is the primary structural component of caveolae and is implicated in the processes of vesicular transport, cholesterol balance, transformation, and tumorigenesis. Despite an abundance of data suggesting that Cav-1 has transformation suppressor properties both in vitro and in vivo, Cav-1 is expressed at increased levels in human prostate cancer. To investigate the role of Cav-1 in prostate cancer onset and progression, we interbred Cav-1(-/-) null mice with a TRAMP (transgenic adenocarcinoma of mouse prostate) model that spontaneously develops advanced prostate cancer and metastatic disease. We found that, although the loss of Cav-1 did not affect the appearance of minimally invasive prostate cancer, its absence significantly impeded progression to highly invasive and metastatic disease. Inactivation of one (+/-) or both (-/-) alleles of Cav-1 resulted in significant reductions in prostate tumor burden, as well as decreases in regional lymph node metastases. Moreover, further examination revealed decreased metastasis to distant organs, such as the lungs, in TRAMP/Cav 1(-/-) mice. Utilizing prostate carcinoma cell lines (C1, C2, and C3) derived from TRAMP tumors, we also showed a positive correlation between Cav-1 expression and the ability of these cells to form tumors in vivo. Furthermore, down regulation of Cav-1 expression in these cells, using a small interfering RNA approach, significantly reduced their tumorigenic and metastatic potential. Mechanistically, we showed that loss or down-regulation of Cav-1 expression results in increased apoptosis, with increased prostate apoptosis response factor 4 and PTEN levels in Cav-1(-/-) null prostate tumors. Our current findings provide the first in vivo molecular genetic evidence that Cav-1 does indeed function as a tumor promoter during prostate carcinogenesis, rather than as a tumor suppressor. PMID- 15802274 TI - Phosphorylation by the DHIPK2 protein kinase modulates the corepressor activity of Groucho. AB - Groucho function is essential for Drosophila development, acting as a corepressor for specific transcription factors that are downstream targets of various signaling pathways. Here we provide evidence that Groucho is phosphorylated by the DHIPK2 protein kinase. Phosphorylation modulates Groucho corepressor activity by attenuating its protein-protein interaction with a DNA-bound transcription factor. During eye development, DHIPK2 modifies Groucho activity, and eye phenotypes generated by overexpression of Groucho differ depending on its phosphorylation state. Moreover, analysis of nuclear extracts fractionated by column chromatography further shows that phospho-Groucho associates poorly with the corepressor complex, whereas the unphosphorylated form binds tightly. We propose that Groucho phosphorylation by DHIPK2 and its subsequent dissociation from the corepressor complex play a key role in relieving the transcriptional repression of target genes regulated by Groucho, thereby controlling cell fate determination during development. PMID- 15802275 TI - Deep vein thrombosis in cancer: the scale of the problem and approaches to management. AB - Patients with cancer have long been recognised to be at high risk of venous thromboembolism (VTE), although the condition remains under diagnosed and under treated in these patients. As a consequence, the morbidity and mortality due to deep venous thrombosis and pulmonary embolism remains unacceptably high in this group. Furthermore, the management of VTE in the presence of malignancy is complex, due both to the effects of the cancer itself and its treatments. Conventional long-term management of VTE involves the use of vitamin K antagonists (VKAs), such as warfarin, to reduce the risk of recurrence. However, this approach is associated with a range of practical difficulties including the need for regular laboratory monitoring, the potential for drug interactions, in addition to the risk of treatment resistance and bleeding in patients with cancer. Recent research indicates that the use of low molecular weight heparin (LMWH) therapy instead of VKAs may be beneficial in these patients. In particular, evidence from a large clinical trial of the LMWH dalteparin indicates that this agent offers an effective alternative to VKAs in the long-term management of VTE, that is free from the practical problems associated with the use of VKAs and without increasing the risk of bleeding. PMID- 15802276 TI - Bisphosphonates in breast cancer. AB - Breast cancer is the leading type of cancer among women, and bone metastases are common in patients with breast cancer, affecting more than half of all patients with advanced disease. Bisphosphonates are the current standard of care for preventing skeletal complications associated with bone metastases. Clinical trials investigating the benefit of bisphosphonate therapy have used a composite end point defined as a skeletal-related event (SRE) or bone event, which typically includes pathologic fracture, spinal cord compression, radiation or surgery to bone, and hypercalcaemia of malignancy. Bisphosphonates significantly reduced the incidence of these events. Zoledronic acid, pamidronate, clodronate and ibandronate have demonstrated efficacy compared with placebo. Zoledronic acid has also been compared with another active bisphosphonate (i.e. pamidronate) and shown by multiple event analysis to be significantly more effective at reducing the risk of SREs. Bisphosphonates effectively reduce and prevent skeletal complications in patients with bone metastases from breast cancer. Preclinical data suggest that bisphosphonates have antitumour effects. Bisphosphonates may also be of use in the adjuvant setting. PMID- 15802277 TI - Clinical course of thoracic cancers in Hodgkin's disease survivors. AB - BACKGROUND: Hodgkin's disease survivors have a high risk of subsequently developing thoracic cancers. Our goal was to evaluate the prognosis and treatment outcomes of thoracic cancers after Hodgkin's disease. PATIENTS AND METHODS: Thirty-three patients treated for Hodgkin's disease at Harvard-affiliated hospitals subsequently developed small-cell lung carcinoma, non-small-cell lung carcinoma (NSCLC) or mesothelioma. Information was obtained from medical records about the initial treatment for Hodgkin's disease, any salvage therapy, smoking history, and the stage, histology, treatment and survival for thoracic cancers. RESULTS: Of the 33 patients, 29 (88%) had a history of radiotherapy to the thorax, 17 (52%) had received alkylating chemotherapy, and 24 (73%) had a known history of smoking. The median time between diagnosis of Hodgkin's disease and diagnosis of thoracic cancer was 17.3 years (range 1.2-27.9 years). Among patients with NSCLC and a known stage, 85% presented with stage III or stage IV disease. Among patients whose treatment details were available, 40% underwent surgery, 40% received radiotherapy and 65% received chemotherapy. The median survival was 9 months (range 1-47 months). CONCLUSIONS: Most patients with thoracic cancers after Hodgkin's disease have a history of exposure to risk factors and present at an advanced stage. Patients with thoracic cancers after Hodgkin's disease have a poor survival. PMID- 15802278 TI - Worse survival for TP53 (p53)-mutated breast cancer patients receiving adjuvant CMF. AB - BACKGROUND: TP53 has been described as a prognostic factor in many malignancies, including breast cancer. Whether it also might be a predictive factor with reference to chemo- and endocrine therapy is more controversial. PATIENTS AND METHODS: We investigated relapse-free (RFS), breast cancer-corrected (BCCS) and overall survival (OS) related to TP53 status in node-positive breast cancer patients that had received polychemotherapy [cyclophosphamide, methotrexate, 5 fluorouracil (CMF)] and/or endocrine therapy (tamoxifen). Sequence analyses of the whole TP53 coding region was performed in 376 patients operated on for primary breast cancer with axillary lymph node metastases between 1984 and 1989 (median follow-up time 84 months). RESULTS: TP53 mutations were found in 105 patients (28%). We found 90 (82%) of the 110 mutations in the more frequently analysed exons 5-8, while the other 20 (18%) were located in exons 3-4 and 9-10, respectively. Univariate analyses showed TP53 to be a significant prognostic factor with regard to RFS, BCCS and OS in patients who received adjuvant CMF. CONCLUSIONS: TP53 mutations might induce resistance to certain modalities of breast cancer therapy. Sequence-determined TP53 mutation was of negative prognostic value in the total patient population and in the CMF treated patients. PMID- 15802279 TI - Cyclin E expression in breast cancer: predicting germline BRCA1 mutations, prognosis and response to treatment. AB - BACKGROUND: Elevated levels of the cell cycle protein cyclin E, and low levels of its inhibitor, p27(Kip1), have been associated with a poor prognosis following breast cancer. Some studies have found that germline mutations in the breast cancer susceptibility gene, BRCA1, are also associated with an inferior survival rate. The relationship between cyclin E/p27(Kip1) levels, BRCA1 status and outcome has not been studied in detail. PATIENTS AND METHODS: We analyzed a historical cohort of 288 Ashkenazi Jewish women who were diagnosed with breast cancer between 1980 and 1995 and were previously tested for BRCA1/2 mutations. Protein levels of cyclin E and p27(Kip1) were assessed by immunohistochemistry. Breast cancer-specific survival (BCSS) was the main outcome measured. RESULTS: The median follow-up was 8 years. Thirty tumors carried germline BRCA1 mutations. These tumors were more likely to have high cyclin E protein levels [odds ratio (OR) 9.5; P <0.001] and low p27(Kip1) protein levels (OR 2.8; P=0.03) than tumors from patients without BRCA1/2 mutations. High cyclin E expression level was the strongest predictor of BRCA1 germline mutations (multivariate OR 4.7; P=0.004). On univariate analysis, high cyclin E protein levels [relative risk (RR) 2.6; P <0.001] and low p27(Kip1) protein levels (RR 2.3; P=0.006) were significant prognostic factors for a poorer BCSS. In Cox multivariate models, high cyclin E levels remained an independent indicator of poor outcome only in the subgroup of patients who did not receive chemotherapy (P=0.002). CONCLUSIONS: In this ethnically restricted cohort, a high level of cyclin E is a characteristic of BRCA1-related breast cancer, and is a marker of poor prognosis following breast cancer, particularly in the absence of adjuvant chemotherapy. PMID- 15802281 TI - Risk profile and outcome in Hodgkin's lymphoma: is obesity beneficial? PMID- 15802280 TI - Report of a European consensus workshop to develop recommendations for the optimal use of (90)Y-ibritumomab tiuxetan (Zevalin) in lymphoma. AB - BACKGROUND: Non-Hodgkin's lymphoma (NHL) comprises a group of related haematological malignancies, predominantly of B-cell origin, which have been described as indolent or aggressive according to their clinical course. Standard treatment for indolent NHL consists of conventional chemotherapy, but, although long-term remissions may occur, most patients will die of their disease. Radioimmunotherapy (RIT) is a novel modality for treating indolent NHL, using monoclonal antibodies to target tumour cells with systemic, low-dose radiation. (90)Y-Ibritumomab tiuxetan (Zevalin); Schering AG, Berlin, Germany), the first RIT approved for use in relapsed/refractory indolent NHL, comprises the murine anti-CD20 monoclonal antibody ibritumomab, covalently linked to the high-energy beta-emitter, yttrium-90, by the chelator, tiuxetan. MATERIALS AND METHODS: A multidisciplinary consensus workshop of European clinicians who had taken part in clinical trials of (90)Y-ibritumomab tiuxetan was convened to develop recommendations for the clinical preparation and administration of (90)Y ibritumomab tiuxetan in Europe. The workshop was held in anticipation of European Medicines Agency approval of this agent, which was gained in 2004 for adult patients with rituximab-relapsed or refractory CD20(+) follicular B-cell NHL. RESULTS AND CONCLUSIONS: This article summarises the consensus recommendations developed for hemato-oncologists. PMID- 15802282 TI - Oncology for medical students: a new ESO educational avenue. PMID- 15802283 TI - Liver does not care about age. PMID- 15802284 TI - Carboplatin plus gemcitabine in patients with inoperable or metastatic pancreatic cancer: a phase II multicenter study by the Hellenic Cooperative Oncology Group. AB - BACKGROUND: In the present phase II multicenter study, we assessed the efficacy and tolerability of the combination of gemcitabine and carboplatin in patients with advanced pancreatic cancer. PATIENTS AND METHODS: Patients with previously untreated, locally advanced or metastatic pancreatic cancer were treated with gemcitabine 800 mg/m(2) on days 1 and 8 and carboplatin at an AUC of 4 on day 8 of a 3-week cycle, for a total of six cycles. Primary end points were response rate and clinical benefit; secondary end points were, survival, time to progression (TTP) and toxicity. RESULTS: A total of 50 patients were enrolled in the study, 47 of whom were eligible for treatment. The median age was 63 years (range 34-76) and the median Karnofsky performance status (PS) was 80%. Patients received a median of six cycles (range 1-11). Among 35 patients evaluable for response, eight (17%) achieved partial response; 15 (32%) and 12 (25%) patients had stable and progressive disease, respectively. The median overall survival was 7.4 months; the median TTP was 4.4 months and the 1-year survival was 28%. The observed clinical benefit response was remarkable. After the second cycle of chemotherapy, 21 of 31 (68%) patients experienced pain improvement and reduced analgesic consumption. At the same time, 35% and 56% of our patients significantly improved their Karnofsky PS and weight, respectively. Overall, the treatment was well tolerated. The most common grade 3-4 toxicities were hematological, including 8% anemia, 6% neutropenia and 13% thrombocytopenia. CONCLUSIONS: The combination of gemcitabine plus carboplatin is a moderately active treatment for patients with locally advanced and metastatic pancreatic cancer. This regimen has an acceptable toxicity profile and provides a significant clinical benefit, and hence warrants further investigation. PMID- 15802285 TI - Density guided importance sampling: application to a reduced model of protein folding. AB - MOTIVATION: Monte Carlo methods are the most effective means of exploring the energy landscapes of protein folding. The rugged topography of folding energy landscapes causes sampling inefficiencies however, particularly at low, physiological temperatures. RESULTS: A hybrid Monte Carlo method, termed density guided importance sampling (DGIS), is presented that overcomes these sampling inefficiencies. The method is shown to be highly accurate and efficient in determining Boltzmann weighted structural metrics of a discrete off-lattice protein model. In comparison to the Metropolis Monte Carlo method, and the hybrid Monte Carlo methods, jump-walking, smart-walking and replica-exchange, the DGIS method is shown to be more efficient, requiring no parameter optimization. The method guides the simulation towards under-sampled regions of the energy spectrum and recognizes when equilibrium has been reached, avoiding arbitrary and excessively long simulation times. AVAILABILITY: Fortran code available from authors upon request. CONTACT: m.j.parker@leeds.ac.uk. PMID- 15802286 TI - A comprehensive and non-redundant database of protein domain movements. AB - MOTIVATION: The current DynDom database of protein domain motions is a user created database that suffers from selectivity and redundancy. The aim of the analysis presented here was to overcome both these limitations and to produce both a comprehensive and a non-redundant description of domain movements from structures stored in the current protein data bank. RESULTS: A multi-step procedure is applied that starts with grouping proteins in the structural databank into families based on sequence similarity. Multiple sequence alignment, conformational clustering and a dimensional clustering method based on the Gram Schmidt algorithm are applied to members of each family to remove dynamic redundancy in their domain movements. Representative domain movements are described in terms of domains, hinge axes and hinge-bending residues using the DynDom program. The results show that within an average family of 11.5 members, there are on average only 1.31 different domain movements indicating a high redundancy in the movements these structures represent. This verifies earlier findings that domain movements are usually highly controlled. Despite the removal of this considerable redundancy, the process has resulted in double the number of domain movements stored in the user-created database. The data are organized in a relational database with a web-interface. AVAILABILITY: The database can be browsed and searched at http://www.cmp.uea.ac.uk/dyndom CONTACT: sjh@cmp.uea.ac.uk. PMID- 15802287 TI - A stochastic differential equation model for quantifying transcriptional regulatory network in Saccharomyces cerevisiae. AB - MOTIVATION: The explosion of microarray studies has promised to shed light on the temporal expression patterns of thousands of genes simultaneously. However, available methods are far from adequate in efficiently extracting useful information to aid in a greater understanding of transcriptional regulatory network. Biological systems have been modeled as dynamic systems for a long history, such as genetic networks and cell regulatory network. This study evaluated if the stochastic differential equation (SDE), which is prominent for modeling dynamic diffusion process originating from the irregular Brownian motion, can be applied in modeling the transcriptional regulatory network in Saccharomyces cerevisiae. RESULTS: To model the time-continuous gene-expression datasets, a model of SDE is applied to depict irregular patterns. Our goal is to fit a generalized linear model by combining putative regulators to estimate the transcriptional pattern of a target gene. Goodness-of-fit is evaluated by log likelihood and Akaike Information Criterion. Moreover, estimations of the contribution of regulators and inference of transcriptional pattern are implemented by statistical approaches. Our SDE model is basic but the test results agree well with the observed dynamic expression patterns. It implies that advanced SDE model might be perfectly suited to portray transcriptional regulatory networks. AVAILABILITY: The R code is available on request. CONTACT: cykao@csie.ntu.edu.tw SUPPLEMENTARY INFORMATION: http://www.csie.ntu.edu.tw/~b89x035/yeast/ PMID- 15802288 TI - GandrKB--ontological microarray annotation and visualization. AB - SUMMARY: The Gandr (gene annotation data representation) knowledgebase is an ontological framework for laboratory-specific gene annotation. Gandr uses Protege 2000 for editing, querying and visualizing microarray data and annotations. Genes can be annotated with provided, newly created or imported ontological concepts. Annotated genes can inherit assigned concept properties and can be related to each other. The resulting knowledgebase can be visualized as interactive network of nodes and edges representing genes and their functional relationships. This allows for immediate and associative gene context exploration. Ontological query techniques allow for powerful data access. PMID- 15802289 TI - Non-invasive pulsatile arterial pressure and stroke volume changes from the human finger. AB - In this paper we review recent developments in the methodology of non-invasive finger arterial pressure measurement and the information about arterial flow that can be obtained from it. Continuous measurement of finger pressure based on the volume-clamp method was introduced in the early 1980s both for research purposes and for clinical medicine. Finger pressure tracks intra-arterial pressure but the pressure waves may differ systematically both in shape and magnitude. Such bias can, at least partly, be circumvented by reconstruction of brachial pressure from finger pressure by using a general inverse anti-resonance model correcting for the difference in pressure waveforms and an individual forearm cuff calibration. The Modelflow method as implemented in the Finometer computes an aortic flow waveform from peripheral arterial pressure by simulating a non-linear three element model of the aortic input impedance. The methodology tracks fast changes in stroke volume (SV) during various experimental protocols including postural stress and exercise. If absolute values are required, calibration against a gold standard is needed. Otherwise, Modelflow-measured SV is expressed as change from control with the same precision in tracking. Beat-to-beat information on arterial flow offers important and clinically relevant information on the circulation beyond what can be detected by arterial pressure. PMID- 15802290 TI - Insulin-stimulated plasma membrane association and activation of Akt2, aPKC zeta and aPKC lambda in high fat fed rodent skeletal muscle. AB - Several recent reports using cell lines have suggested that both Akt and atypical protein kinase C (aPKC) zeta/lambda are translocated to the plasma membrane (PM) in response to insulin. However, it has yet to be determined in skeletal muscle whether: (1) insulin increases PM-associated Akt2, aPKC zeta and/or lambda protein concentration, (2) the activity of these kinases is altered by insulin at the PM, and (3) high fat feeding alters the insulin-stimulated PM concentration and/or activity of Akt2 and aPKC zeta/lambda. Sprague-Dawley rats were randomly assigned to either normal (n=16) or high fat (n=16) dietary groups. Following a 12 week dietary period, animals were subjected to hind limb perfusions in the presence (n=8 per group) or absence (n=8 per group) of insulin. In normal skeletal muscle, total PI3-kinase, Akt2 and aPKC zeta/lambda activities were increased by insulin. PM-associated aPKC zeta and lambda, and aPKC zeta/lambda activity, but not Akt2 or Akt2 activity, were increased by insulin in normal muscle. High fat feeding did not alter total skeletal muscle Akt2, aPKC zeta or aPKC lambda protein concentration. Insulin-stimulated total PI3-kinase, Akt2 and aPKC zeta/lambda activities were reduced in the high fat fed animals. Insulin stimulated PM aPKC zeta, aPKC lambda, aPKC zeta/lambda activity and GLUT4 protein concentration were also reduced in high fat fed animals. These findings suggest that in skeletal muscle, insulin stimulates translocation of aPKC zeta and lambda, but not Akt2, to the PM. In addition, high fat feeding impairs insulin stimulated activation of total aPKC zeta/lambda and Akt2, as well as PM association and activation of aPKC zeta and lambda. PMID- 15802291 TI - Sensory transduction of pulmonary reactive oxygen species by capsaicin-sensitive vagal lung afferent fibres in rats. AB - The mechanisms of sensory transduction of pulmonary reactive oxygen species (ROS) by capsaicin-sensitive vagal lung afferent fibres are unclear. To investigate the role of transient receptor potential vanilloid 1 (TRPV1) receptors and P2X purinoceptors in this sensory transduction, we recorded fibre activity (FA) from 132 fibres of this type in 132 anaesthetized and ventilated rats. Airway challenge of aerosolized H2O2 (0, 0.2 and 0.4%) produced a concentration dependant fibre stimulation. The fibre responses to 0.4% H2O2 were attenuated by dimethylthiourea (a hydroxyl radical (.OH) scavenger; change in fibre activity (DeltaFA), -55+/-9%) or deferoxamine (an iron-chelator that prevents formation of .OH; DeltaFA, -59+/-9%), were prevented by catalase (an enzyme catalysing H2O2; DeltaFA, -96+/-3%) and were unaffected by the vehicle for dimethylthiourea, iron saturated deferoxamine or heat-inactivated catalase. The fibre responses to 0.4% H2O2 were attenuated by capsazepine (a TRPV1 receptor antagonist; DeltaFA, -39+/ 9%) or iso-pyridoxalphosphate-6-azophenyl-2',5'-disulphonate (iso-PPADS, a P2X receptor antagonist; DeltaFA, -51+/-9%), were further reduced by capsazepine and iso-PPADS in combination (DeltaFA, -70+/-13%), and were unaltered by their vehicles. The fibre responses to cigarette smoke (20 ml), an irritant that generates ROS, were attenuated by dimethylthiourea (DeltaFA, -61+/-9%) or capsazepine and iso-PPADS in combination (DeltaFA, -67+/-9%). These results suggest that both the TRPV1 and P2X receptors mediate the sensory transduction of ROS, especially H2O2 and .OH, by capsaicin-sensitive vagal lung afferent fibres. PMID- 15802292 TI - Preserved contribution of nitric oxide to baseline vascular tone in deconditioned human skeletal muscle. AB - Deconditioning is a risk factor for cardiovascular disease. Exercise reduces this risk, possibly by improving the vascular endothelial nitric oxide (NO) pathway. The effect of deconditioning on the NO pathway is largely unknown. This study was designed to assess baseline NO availability in the leg vascular bed after extreme, long-term deconditioning (spinal cord-injured individuals, SCI) as well as after moderate, short-term deconditioning (4 weeks of unilateral lower limb suspension, ULLS). For this purpose, seven SCI were compared with seven matched controls. Additionally, seven healthy subjects were studied pre- and post-ULLS. Leg blood flow was measured by venous occlusion plethysmography at baseline and during infusion of 5 incremental dosages of N(G)-monomethyl-L-arginine (L-NMMA) into the femoral artery. Sodium nitroprusside (SNP) was infused to test vascular responsiveness to NO. Baseline leg vascular resistance tended to be higher in SCI compared with controls (37+/-4 versus 31+/-2 arbitrary units (AU), P=0.06). Deconditioning altered neither the vasoconstrictor response to L-NMMA (increase in resistance in SCI versus controls: 102+/-33% versus 69+/-9%; pre- versus post ULLS: 95+/-18% versus 119+/-15%), nor the vascular responsiveness to NO. In conclusion, two human in vivo models of deconditioning show a preserved baseline NO availability in the leg skeletal muscle vascular bed. PMID- 15802293 TI - Computational modelling of H+-coupled peptide transport via human PEPT1. AB - H+-coupled peptide transporter 1 (PEPT1) mediates the transport of small peptides and peptide-like drugs in a pH- and voltage-dependent manner. Here, we investigated the transport mechanisms of PEPT1 for neutral and charged substrates by experimental studies and computational simulation. Uptake studies revealed that the Michaelis-Menten constant (Km) of glycylsarcosine (Gly-Sar), a neutral substrate, decreased with a fall in pH from 7.4 to 5.5, but at pH 5.0, the Km increased again. In contrast, the Km value of an anionic substrate, ceftibuten, declined steadily with decreasing pH. Based on these findings and information from the literature, we hypothesized the transport mechanisms in which (1) H+ binds to not only the H+-binding site, but also the substrate-binding site; and (2) H+ at the substrate-binding site inhibits the interaction of neutral and cationic substrates, but is necessary for that of anionic substrates. To validate these hypotheses, a computational model was constructed and various properties of substrate transport by PEPT1 were simulated. Our model reproduced the voltage dependence, hyperbolic saturation and bell-shaped pH-profile of Gly-Sar transport. Moreover, the various transport properties of negatively and positively charged substrates were also reconstructed. These findings indicated that the inferred mechanisms are able to sufficiently interpret the transport of both neutral and charged substrates by PEPT1. PMID- 15802294 TI - Post-stimulus potentiation of transmission in pelvic ganglia enhances sympathetic dilatation of guinea-pig uterine artery in vitro. AB - Vasodilatation produced by stimulation of preganglionic neurones in lumbar and sacral pathways to pelvic ganglia was studied using an in vitro preparation of guinea-pig uterine artery and associated nerves in a partitioned bath allowing selective drug application to the ganglia or artery. Arterial diameter was monitored using real time video imaging. Vasodilatations produced by hypogastric nerve stimulation (HN; 300 pulses, 10 Hz) were significantly larger and longer in duration than with pelvic nerve stimulation (N = 18). Stimulation of ipsilateral lumbar splanchnic nerves or ipsilateral third lumbar ventral roots also produced prolonged vasodilatations. Blockade of ganglionic nicotinic receptors (0.1-1 mM hexamethonium) delayed the onset and sometimes reduced the peak amplitude of dilatations, but slow dilatations persisted in 16 of 18 preparations. These dilatations were not reduced further by 3 microM capsaicin applied to the artery and ganglia, or ganglionic application of 1 microM hyoscine, 30-100 microM suramin or 10 microM CNQX. Dilatations were reduced slightly by ganglionic application of NK1 and NK3 receptor antagonists (SR140333, SR142801; 1 microM), but were reduced significantly by bathing the ganglia in 0.5 mM Ca2+ and 10 mM Mg2+. Intracellular recordings of paracervical ganglion neurones revealed fast excitatory postsynaptic potentials (EPSPs) in all neurones on HN stimulation (300 pulses, 10 Hz), and slow EPSPs (3-12 mV amplitude) in 25 of 37 neurones. Post stimulus action potential discharge associated with slow EPSPs occurred in 16 of 37 neurones (firing rate 9.4 +/- 1.5 Hz). Hexamethonium (0.1-1 mM) abolished fast EPSPs. Hexamethonium and hyoscine (1 microM) did not reduce slow EPSPs and associated post-stimulus firing in identified vasodilator neurones (with VIP immunoreactivity) or non-vasodilator paracervical neurones. These results demonstrate a predominantly sympathetic origin of autonomic pathways producing pelvic vasodilatation in females. Non-cholinergic mediators of slow transmission in pelvic ganglia produce prolonged firing of postganglionic neurones and long lasting dilatations of the uterine artery. This mechanism would facilitate maintenance of pelvic vasodilatation on stimulation of preganglionic neurones during sexual activity. PMID- 15802295 TI - Inhibitory and excitatory intracortical circuits across the human sleep-wake cycle using paired-pulse transcranial magnetic stimulation. AB - Studies using single-pulse transcranial magnetic stimulation (TMS) have shown that excitability of the corticospinal system is systematically reduced in natural human sleep as compared to wakefulness with significant differences between sleep stages. However, the underlying excitatory and inhibitory interactions on the corticospinal system across the sleep-wake cycle are poorly understood. Here, we specifically asked whether in the motor cortex short intracortical inhibition (SICI) and facilitation (ICF) can be elicited at all in sleep using the paired-pulse TMS protocol, and if so, how SICI and ICF vary across sleep stages. We studied 28 healthy subjects at interstimulus intervals of 3 ms (SICI) and 10 ms (ICF), respectively. Magnetic stimulation was performed over the hand area of the motor cortex using a focal coil and evoked motor potentials were recorded from the contralateral first dorsal interosseus muscle (1DI). Relevant data was obtained from 13 subjects (NREM 2: n=7; NREM 3/4: n=7; REM: n=7). Results show that both SICI and ICF were present in NREM sleep. SICI was significantly enhanced in NREM 3/4 as compared to wakefulness and all other sleep stages whereas in NREM 2 neither SICI nor ICF differed from wakefulness. In REM sleep SICI was in the same range as in wakefulness, but ICF was entirely absent. These results in humans support the hypothesis derived from animal experiments which suggests that intracortical inhibitory mechanisms are involved in the control of neocortical pyramidal cells in NREM and REM sleep, but along different intraneuronal circuits. Further, our findings suggest that cortical mechanisms may additionally contribute to the inhibition of spinal motoneurones in REM sleep. PMID- 15802296 TI - An acute decrease in TCA cycle intermediates does not affect aerobic energy delivery in contracting rat skeletal muscle. AB - We tested the hypothesis that an acute decrease in muscle TCA cycle intermediates during contraction would compromise aerobic energy delivery. Male Wistar rats were anaesthetized and the gastrocnemius-plantaris-soleus (GPS) muscle complex from one leg was isolated and perfused with a red cell medium containing either saline (Con) or cycloserine (Cyclo; 0.05 mg g-1), an inhibitor of alanine aminotransferase (AAT). After 1 h of perfusion, the GPS muscle was either snap frozen (Con-Rest, n=11; Cyclo-Rest, n=9) or stimulated to contract for 10 min (1 Hz, 0.3 ms, 2 V) with blood flow fixed at 30 ml min-1 (100 g)-1 and then snap frozen (Con-Stim, n=10; Cyclo-Stim, n=10). Maximal AAT activity was>80% lower (P<0.001) in both Cyclo-treated groups (Rest: 0.61+/-0.02; Stim: 0.63+/-0.01 mmol (kg wet wt)-1 min-1; mean+/-s.e.m.) compared to Con (Rest: 3.56+/-0.16; Stim: 3.92+/-0.29). The sum of five measured TCAI (SigmaTCAI) was reduced by 23% in Cyclo-Rest versus Con-Rest but this was not different (P=0.08). However, after 10 min of contraction, the SigmaTCAI was 25% lower (P=0.006) in Cyclo-Stim compared to Con-Stim (1.88+/-0.15 versus 2.48+/-0.11 mmol (kg dry wt)-1). Despite the acute decrease in TCAI after Cyclo treatment, the contraction-induced changes in markers of non-oxidative energy provision (phosphocreatine, ATP and lactate) and the decline in tension after 10 min of stimulation were similar compared to Con. These data do not support the hypothesis that the total muscle concentration of TCAI is causally linked to the rate of mitochondrial respiration during contraction. PMID- 15802297 TI - Aurora-A/STK15 T+91A is a general low penetrance cancer susceptibility gene: a meta-analysis of multiple cancer types. AB - STK15 (Aurora-A) is a serine/threonine kinase involved in mitotic chromosomal segregation. A genetic variant in STK15 T+91A (resulting in the amino acid substitution F31I) is associated with increased aneuploidy in colon tumors and cell transformation in vitro. Since this polymorphism plays a role in mitotic control-a process critical for all cancer types-we conducted association analyses for risk of cancer development of the colon, breast, prostate, skin, lung and esophagus in 10 independent case-control populations. We carried out a meta analysis of these 10 case-control studies together with 5 additional published studies for a total of 9549 cases of breast, colon, ovarian, prostate, lung, esophageal and non-melanoma skin cancer and 8326 population or hospital-based controls. Meta-analysis of three colorectal cancer studies showed an increased risk in T+91A homozygotes (OR=1.50; 95% CI of 1.14-1.99). Meta-analysis of four breast cancer studies showed increased risk for T+91A homozygotes (OR=1.35, 95% CI of 1.12-1.64). The results of the multiple cancer type meta-analysis for all 15 studies combined were significant for cancer risk in both homozygotes and heterozygotes. The T+91A heterozygotes show an OR of 1.10 (95% CI of 1.03-1.18, P value=0.006) and the T+91A homozygotes show an OR of 1.40 (95% CI of 1.22-1.59, P value<0.001) for cancer risk. These results confirm that the STK15 T+91A variant is a low penetrance cancer susceptibility allele affecting multiple cancer types, and provide genetic evidence from large-scale human population studies that genetic stability at the chromosome level is an important determinant of cancer susceptibility. The data also underline the advantages of comparative association studies involving study populations from different ethnic groups for determination of disease risk. PMID- 15802298 TI - Selected DNA repair polymorphisms and gastric cancer in Poland. AB - Impaired DNA repair capacity may adversely affect cancer risk, particularly in subjects exposed to DNA damaging carcinogens, as found in tobacco smoke, or among subjects deficient for protective factors, as found in fruits and vegetables. We studied tobacco use, fruit and vegetable intake, and common non-synonymous single nucleotide polymorphisms in four DNA repair genes in relation to gastric cancer risk, in a population-based, case-control study of 281 incident gastric cancer cases and 390 controls, in Warsaw, Poland. Multivariate logistic regression analysis was performed to calculate odds ratios (OR) and 95% confidence intervals (CI). Increased risks of gastric cancer were found for smokers (OR=3.1, CI=1.9 5.1 for pack-years>or=40 versus never smokers) and subjects with low fruit intake (OR=2.2, CI=1.3-3.6 for 1st versus 4th quartile); risk associated with vegetable intake was not statistically significant. Allele frequencies among the controls were consistent with those previously reported for the 5 polymorphisms studied: XRCC1-Arg399Gln, XPD-Lys751Gln, MGMT-Ile143Val, Leu84Phe, and XRCC3-Thr241Met. None of the studied polymorphisms were independently associated with gastric cancer risk. Smoking-associated risks, however, were greatest for carriers of the XRCC1-399 ArgArg genotype (Pinteraction=0.004). Risks associated with low intake of fruits or vegetables tended to be modified by selected polymorphisms in XRCC1, XPD and MGMT (Pinteraction=0.1-0.2). Risk modification was not found for the other repair polymorphisms. Selected DNA repair polymorphisms did not have independent effects on gastric cancer risk; however, they may modify smoking- and probably diet-related risks for this disease. These results need replication in larger epidemiological studies of gastric cancer. PMID- 15802299 TI - Cloudy apple juice decreases DNA damage, hyperproliferation and aberrant crypt foci development in the distal colon of DMH-initiated rats. AB - Clear (CleA) and cloudy (CloA) apple juices containing different amounts of analyzed procyanidins and pectin were investigated for preventive effects of colon cancer and underlying molecular mechanisms in F344 rats given intraperitoneal injections of 1,2-dimethylhydrazine (DMH; 20 mg/kg body wt) once a week for 4 weeks. Rats received either water (Cont), CleA or CloA (ad libitum) for 7 weeks starting 1 week before the first DMH injection. CloA inhibited DMH induced genotoxic damage in mucosa cells of the distal colon compared with Cont as investigated by single-cell microgel electrophoresis assay. The mean tail intensity in mucosa cells of DMH-treated controls (Cont/DMH: 6.1+/-0.9%) was significantly reduced by CloA (2.4+/-0.8%; P<0.01) but not by CleA intervention (4.1+/-1.2%; P>0.05). The crypt cell proliferation index induced by DMH (Cont/NaCl: 10.0+/-0.7%; Cont/DMH: 19.9+/-1.0%; P<0.001) was significantly decreased by CleA (15.7+/-0.7%; P<0.001) and CloA intervention (11.9+/-0.4%; P<0.001). CloA but not CleA significantly reduced the number of large aberrant crypt foci (ACF) consisting of more than four aberrant crypts (AC) (Cont/DMH: 37.4+/-5.4; CleA/DMH: 32.8+/-4.4, P>0.05; CloA/DMH: 18.8+/-2.5 ACF; P<0.05) and the overall mean ACF size in the distal colon (Cont/DMH: 2.31+/-0.09; CleA/DMH: 2.27+/-0.05; CloA/DMH: 2.04+/-0.03 AC/ACF; P<0.05). After treatment with DMH and/or apple juices there were no changes in transcript levels of colonic cyclooxygenase isoforms (COX-1, COX-2) or glutathione-associated enzymes (GST-M2, gamma-GCS, GST-P), the splenocyte natural killer cell activity and plasma antioxidant status. However, CloA but not CleA prevented the DMH-induced reduction of splenocyte CD4/CD8 (T-helper cells to cytotoxic lymphocytes) ratio. Since both formulations contained comparable concentrations and types of monomeric polyphenols, complex polyphenols or non-polyphenolic compounds, such as pectin might be responsible for the stronger cancer-preventive effect by CloA. PMID- 15802300 TI - NF-kappaB inhibition radiosensitizes Ki-Ras-transformed cells to ionizing radiation. AB - Most cancer cells show resistance to ionizing radiation (IR)-induced cell death. Recently, Ki-Ras was reported to be responsible for the increased radioresistance. We report here that inhibition of IR-induced activaton of nuclear transcription factor kappa B (NF-kappaB) but not of either Akt or MAPK kinase (MEK), increased the radiosensitization of Ki-Ras transformed human prostate epithelial 267B1/K-ras cells. Proteosome inhibitor-1 (Pro1) reduced NF kappaB activation, and this inhibition was accompanied by increased levels of cytoplasmic IkappaBalpha and p65/RelA. However, translocation of p50/NF-kappaB1 did not occur on exposure to IR, suggesting the cell-specific involvement of p50 in radiation signaling. Clonogenic cell survival and soft agar assays further confirmed the increased radiosensitivity of 267B1/K-ras cells by proteosome inhibition. In addition, proteosome inhibition enhanced the IR-induced degradation of apoptotic protein caspases 8 and 3, with the level of antiapoptotic protein Bcl-2 being unaffected, suggesting the involvement of an apoptotic process in IR-induced cell death of 267B1/K-ras cells. LY294002 and PD98059, specific inhibitors of phosphatidylinositol-3-kinase (PI3K) and MEK, respectively however, did not affect the radiosensitization. All these results suggest an application of blocking NF-kappaB activation pathway to the development of anticancer therapeutics in IR-induced radiotherapy of Ki-Ras transformed cancer cells. PMID- 15802301 TI - Green tea intake, ACE gene polymorphism and breast cancer risk among Chinese women in Singapore. AB - Experimental and epidemiological data have implicated a potential chemoprotective role of green tea polyphenols and a potential enhancing role of angiotensin II in the development of breast cancer in humans. Angiotensin II is converted from its precursor by angiotensin-converting enzyme (ACE). Women with low-activity genotype of the ACE gene had a reduced risk of breast cancer compared with those possessing high-activity ACE genotype. Experimental data showed that green tea polyphenols could inhibit angiotensin II-induced reactive oxygen species production. We reasoned that if this is one of the mechanisms by which green tea polyphenols protect against human breast cancer, then their effect should be more prominent among women possessing high-activity ACE genotype than women with low activity ACE genotype. In other words, we predict a stronger inverse green tea breast cancer association among the former versus the latter subgroup of women. To test this hypothesis, we conducted a nested case-control study involving 297 incident breast cancer cases and 665 control subjects within the Singapore Chinese Health Study. There was no association between intake frequencies of green tea and risk of breast cancer among all women or those with low-activity ACE genotype. Among women with high-activity ACE genotype, however, intake frequency of green tea was associated with a statistically significant decrease in risk of breast cancer (P for trend=0.039); the odds ratio (95% confidence interval) was 0.33 (0.13-0.82) for women drinking green tea at least monthly and 0.29 (0.10-0.79) for those drinking green tea at least weekly compared with non drinkers. There was a statistically significant interaction effect between green tea intake and ACE genotype on risk of breast cancer (P for interaction=0.01). Black tea intake was unrelated to breast cancer risk irrespective of the ACE genotype. The findings of the present study highlight the importance of genetically determined factors in evaluating the role of green tea intake in the development of breast cancer. PMID- 15802302 TI - Tumorigenesis: the adaptation of mammalian cells to sustained stress environment by epigenetic alterations and succeeding matched mutations. AB - Recent studies indicate that during tumorigenic transformations, cells may generate mutations by themselves as a result of error-prone cell division with participation of error-prone polymerases and aberrant mitosis. These mechanisms may be activated in cells by continuing proliferative and survival signaling in a sustained stress environment (SSE). The paper hypothesizes that long-term exposure to this signaling epigenetically reprograms the genome of some cells and, in addition, leads to their senescence. The epigenetic reprogramming results in: (i) hypermethylation of tumor-suppressor genes involved in the onset of cell cycle arrest, apoptosis and DNA repair; (ii) hypomethylation of proto-oncogenes associated with persistent proliferative activity; and (iii) the global demethylation of the genome and activation of DNA repeats. These epigenetic changes in the proliferating cells associate with their replicative senescence and allow the reprogrammed senescent cells to overcome the cell-cycle arrest and to activate error-prone replications. It is hypothesized that the generation of mutations in the error-prone replications of the epigenetically reprogrammed cells is not random. The mutations match epigenetic alterations in the cellular genome, namely gain of function mutations in the case of hypomethylation and loss of functions in the case of hypermethylation. In addition, continuing proliferation of the cells imposed by signaling in SSE speeds up the natural selection of the mutant cells favoring the survival of the cells with mutations that are beneficial in the environment. In this way, a stress-induced replication of the cells epigenetically reprograms their genome for quick adaptation to stressful environments providing an increased rate of mutations, epigenetic tags to beneficial mutations and quick selection process. In combination, these processes drive the origin of the transformed mammalian cells, cancer development and progression. Support from genomic, biochemical and medical studies for the proposed hypothesis, and its implementations are discussed. PMID- 15802303 TI - Carbohydrate profiling reveals a distinctive role for the C-type lectin MGL in the recognition of helminth parasites and tumor antigens by dendritic cells. AB - Dendritic cells (DCs) are key to the maintenance of peripheral tolerance to self antigens and the orchestration of an immune reaction to foreign antigens. C-type lectins, expressed by DCs, recognize carbohydrate moieties on antigens that can be internalized for processing and presentation. Little is known about the exact glycan structures on self-antigens and pathogens that are specifically recognized by the different C-type lectins and how this interaction influences DC function. We have analyzed the carbohydrate specificity of the human C-type lectin macrophage galactose-type lectin (MGL) using glycan microarray profiling and identified an exclusive specificity for terminal alpha- and beta-linked GalNAc residues that naturally occur as parts of glycoproteins or glycosphingolipids. Specific glycan structures containing terminal GalNAc moieties, expressed by the human helminth parasite Schistosoma mansoni as well as tumor antigens and a subset of gangliosides, were identified as ligands for MGL. Our results indicate an endogenous function for DC-expressed MGL in the clearance and tolerance to self-gangliosides, and in the pattern recognition of tumor antigens and foreign glycoproteins derived from helminth parasites. PMID- 15802304 TI - BAT mAb induces lymphopoiesis in nude mice. AB - The athymic nude mouse provides a powerful tool in the study of human tumors, as it enables growth of human tumors due to deficiencies in T cell functions. However, deficiencies in T cell functions might limit research on efficacy of immune modulators in cancer immunotherapy. BAT mAb mediates its anti-cancer activity through modulation of the immune system that involves both NK and T cells. We analyzed lymphocyte populations in blood 5 and 14 days following the injection of BAT antibody alone or following engraftment of human colon carcinoma cells. Our results demonstrate that BAT injection induced lymphopoiesis in the nude mouse. Percentage of CD3 cells increased up to 24%, CD4 cells up to 20% but no increase was found in CD8 T cells in BAT-injected nude mice. Injection of BAT 12 days post-tumor engraftment propagated CD3, CD4 and CD8 cells seen in the blood 5 days later but not seen in the blood 14 days post-BAT injection. It is possible that this decrease is associated with migration of the lymphocytes from the blood to the tumor sites in the livers. The percentage of CD56-positive NK cells increased (up to 18%) by BAT administration alone or post-tumor injection. The presence of tumors alone did not induce lymphopoiesis in the nude mice. Propagation and lymphopoiesis by BAT mAb might have future clinical implications. PMID- 15802305 TI - 5-Hydroxytryptamine modulates cytokine and chemokine production in LPS-primed human monocytes via stimulation of different 5-HTR subtypes. AB - The neurotransmitter 5-hydroxytryptamine (5-HT), commonly known as serotonin, is released at peripheral sites from activated enterochromaffin cells, mast cells and platelets. In this study we analyzed the biological activity and intracellular signaling of 5-HT in human monocytes. By reverse transcription (RT) and PCR, messenger RNA (mRNA) expression of 5-HT receptor 1E (5-HTR(1E)), 5 HTR(2A), 5-HTR(3), 5-HTR(4) and 5-HTR(7) could be revealed. Functional studies showed that 5-HT modulates the release of IL-1beta, IL-6, IL-8/CXCL8, IL-12p40 and tumor necrosis factor-alpha (TNF-alpha), while it has no effect on the production of IL-18 and IFN-gamma in LPS-stimulated human blood monocytes. Moreover, RT and PCR revealed that 5-HT modulated mRNA levels of IL-6 and IL 8/CXCL8, but did not influence mRNA levels of IL-1beta and TNF-alpha. Pharmacological studies with isotype-selective receptor agonists allowed us to show that 5-HTR(3) subtype up-regulates the LPS-induced production of IL-1beta, IL-6 and IL-8/CXCL8, while it was not involved in TNF-alpha and IL-12p40 secretion. Furthermore, activation of the G(s)-coupled 5-HTR(4) and 5-HTR(7) subtypes increased intracellular cyclic AMP (cAMP) and secretion of IL-1beta, IL 6, IL-12p40 and IL-8/CXCL8, while, on the contrary, it inhibited LPS-induced TNF alpha release. Interestingly, 5-HTR(1) and 5-HTR(2) agonists did not modulate the LPS-induced cytokine production in human monocytes. Our results point to a new role for 5-HT in inflammation by modulating cytokine production in monocytes via activation of 5-HTR(3), 5-HTR(4) and 5-HTR(7) subtypes. PMID- 15802306 TI - CD8+ T cells specific for a potential HLA-A*0201 epitope from Chlamydophila pneumoniae are present in the PBMCs from infected patients. AB - Infection with the common pathogen Chlamydophila pneumoniae (Cpn, previously Chlamydia pneumoniae) has a high prevalence in patients suffering from arteriosclerosis and may trigger or contribute to heart disease. In mice, CD8 positive T cells are critical for the eradication of the infection and the development of immune memory against Cpn. Although several H2-class I epitopes have been described, no HLA-class I-associated peptides from Cpn are known. In order to define HLA-A*0201 epitopes from Cpn, we focused on the bacterial heat shock proteins (HSP) 60 and 70 which are known to be recognized by the immune system. Using epitope prediction, peptide binding studies and peptide-specific CTLs from HLA-A2 transgenic mice, we could define a potential HSP-70-derived epitope. The study of PBMCs from Cpn-infected individuals using fluorescent MHC tetramers revealed that some patients have CD8(+) T cells capable of recognizing the Cpn HSP-70 HLA-A*0201 epitope. Our studies pave the way to the immunomonitoring of the anti-Cpn CTL immune response present in patients suffering from different diseases potentially linked to Cpn or anti-Cpn immunity. PMID- 15802307 TI - NK cell activation and protection occur independently of natural killer T cells during Trypanosoma cruzi infection. AB - Natural killer T (NKT) cells regulate aspects of pro-inflammatory and anti inflammatory responses and contribute to the control of infections and chronic inflammatory diseases. During Trypanosoma cruzi infection both NKT cells and NK cells are critical to the protective response. How NKT cells interact and possibly regulate NK cells during infections remains uncertain. In vivo studies have demonstrated that specific activation of NKT cells with alpha galactosylceramide (alpha-GalCer) leads to NK cell activation. These results suggest that during some infections activated NKT cells might regulate NK cell activation and functions. Therefore, using gene-deficient mice that lack NKT cells and antibody-treated mice that lack NK cells, we investigated the interactions of NKT cells and NK cells during experimental T. cruzi infection. We report here that during acute T. cruzi infection spleen and liver NK cell activation and cytolytic activity occur independently of NKT cells. Moreover, NK cell protection occurs independently of NKT cells. In contrast to these results that fail to demonstrate an interdependence, at day 4 of infection the number of liver NK cells is controlled by NKT cells. Thus, during T. cruzi infection, regulation of the number of liver NK cells requires NKT cells, but the activation of NK cells and protection by NK cells does not. The data presented here argue that during infections NK cell activation and protection occur independently of NKT cells. PMID- 15802308 TI - Redistributions of macrophages expressing the macrophage galactose-type C-type lectin (MGL) during antigen-induced chronic granulation tissue formation. AB - Cell surface lectins are known to regulate trafficking of cells in the immune system, yet the role of macrophage galactose-type C-type lectin 1 and 2 (MGL1/2) is poorly understood. In this study, antigen-specific chronic inflammation was induced in a subcutaneous air pouch model in mice, and distribution of cells expressing MGL1/2 was investigated. Azobenzenearsonate-conjugated acetylated BSA, used as an antigen, was introduced into an air pouch of immunized mice, and tissue formation and distribution of MGL1/2-positive cells in the sub-dermal regions was examined. Thickness of the inflammatory tissue and number of MGL1/2 positive cells simultaneously reached the maximum at day 4 and returned to the control level at day 6 or 8. When additional antigenic challenges were given, a chronic granulation tissue, which had two distinct layers, was generated. In the chronic tissue, CD11b-positive/MGL1/2-negative cells were abundant in the area close to the antigenic stimulus, while the area far from the antigenic stimulus was dominated by MGL1/2-positive/CD11b-negative or -low cells. Flow cytometric analyses of isolated cells from the granulation tissue revealed that MGL1/2 positive cells expressed MHC class II at high levels, CD11b at low levels but no CD11c. MGL1/2-positive and -negative fractions were separated from cells in the granulation tissue and a higher level of IL-1alpha messenger RNA than negative populations was detected in the MGL1/2-positive fraction by the semi-quantitative reverse transcription-PCR method. IL-1alpha production by MGL1/2-positive cells was also immunohistochemically detected. Results suggest that MGL1/2-positive cells represent a distinct sub-population of macrophages, having unique functions in the generation and maintenance of granulation tissue induced by antigenic stimuli. PMID- 15802309 TI - Biomechanical model of the xylem vessels in vascular plants. AB - BACKGROUND AND AIMS: The xylem, or water transport system, in vascular plants adopts different morphologies that appear sequentially during growth phases. This paper proposes an explanation of these morphologies based on engineering design principles. METHODS: Using microscopic observations of the different growth stages, an engineering analysis of the xylem vessels as a closed cylinder under internal pressure is carried out adopting pressure vessel design concepts. KEY RESULTS: The analysis suggests that the xylem vessel structural morphology follows the 'constant strength' design principle, i.e. all of the material within the wall of the xylem is loaded equally to its maximum allowable stress capacity, and the amount of material used is therefore systematically minimized. The analysis shows that the different structural designs of the xylem vessel walls (annular, helical, reticulate and pitted) all quantitatively follow the constant strength design principle. CONCLUSIONS: The results are discussed with respect to growth and differentiation. It is concluded that the morphology of the xylem vessel through the different phases of growth seems to follow optimal engineering design principles. PMID- 15802310 TI - Effects of plant leachates from four boreal understorey species on soil N mineralization, and white spruce (Picea glauca) germination and seedling growth. AB - BACKGROUND AND AIMS: Natural regeneration of white spruce (Picea glauca) after disturbance has been reported to be very poor. Here a study was made to determine whether C compounds released from understorey species growing together with white spruce could be involved in this regeneration failure, either by (1) changing soil nutrient dynamics, (2) inhibiting germination, and/or (3) delaying seedling growth. METHODS: Foliage leachates were obtained from two shrubs (Ledum palustre and Empetrum hermaphroditum) and one bryophyte (Sphagnum sp.) with high phenolic compound concentrations that have been reported to depress growth of conifers in boreal forests, and, as a comparison, one bryophyte (Hylocomium splendens) with negligible phenolic compounds. Mineral soil from a white spruce forest was amended with plant leachates to examine the effect of each species on net N mineralization. Additionally, white spruce seeds and seedlings were watered with plant leachates to determine their effects on germination and growth. KEY RESULTS: Leachates from the shrubs L. palustre and E. hermaphroditum contained high phenolic compound concentrations and dissolved organic carbon (DOC), while no detectable levels of C compounds were released from the bryophytes Sphagnum sp. or H. splendens. A decrease in net N mineralization was determined in soils amended with L. palustre or E. hermaphroditum leachates, and this effect was inversely proportional to the phenolic concentrations, DOC and leachate C/N ratio. The total percentage of white spruce germination and the growth of white spruce seedlings were similar among treatments. CONCLUSIONS: These results suggest that the shrubs L. palustre and E. hermaphroditum could negatively affect the performance of white spruce due to a decrease in soil N availability, but not by direct effects on plant physiology. PMID- 15802311 TI - Dynamics of nitrogen uptake and mobilization in field-grown winter oilseed rape (Brassica napus) from stem extension to harvest. II. An 15N-labelling-based simulation model of N partitioning between vegetative and reproductive tissues. AB - BACKGROUND AND AIMS: Oilseed rape (Brassica napus) has often been used as a catch crop to deal with the issue of N leaching, but for this to be effective, prediction of the crop's N uptake capability and N partitioning is required. The aim of this work was to build a compartmental model of N dynamics in oilseed rape, based on the kinetic description of N uptake, partitioning and mobilization in each organ. MODEL: In this study, logistic and exponential equations were fitted to the N relations of each compartment, especially the leaf at each node. Data previously obtained from an 15N-labelling field experiment was used to quantify the partitioning of total N content, the allocation of N taken up and subsequent changes in the sink/source status for endogenous N in each tissue throughout the growth cycle. KEY RESULTS AND CONCLUSIONS: This modelling approach provides a unique tool for the quantitative estimation of cycling of endogenous N in relation to changes in N uptake at the whole-plant level. Furthermore, as oilseed rape is known to release large amounts of N to the soil during spring through leaf loss, this model was used to identify potential methods for improving the N harvest index of the crop. Simulations showed that N content or yield could be improved by 15% by optimizing N transfer from vegetative to reproductive tissues and by reducing the residual %N (DW) in abscised leaves. PMID- 15802312 TI - Birth weight in offspring of mothers with polycystic ovarian syndrome. AB - BACKGROUND: A relationship between reduced fetal growth and the polycystic ovary syndrome (PCOS) has been proposed in girls with PCOS. However, the birth weight in the offspring of PCOS mothers has not been systematically investigated. The aim of this study was to establish the birth weight of newborns of mothers with PCOS and to compare it with a control group of newborns of normal women matched by age and weight at the beginning of pregnancy. METHODS: The birth weight of 47 infants born from singleton pregnancies in women with well-documented PCOS was compared with 180 infants born from singleton pregnancies in healthy controls. RESULTS: The prevalence of small for gestational age (SGA) infants was significantly higher in the PCOS group compared to the control group (12. 8% versus 2.8%, respectively, P<0.02). Moreover, SGA infants born to PCOS mothers were smaller than those born to control mothers (P<0.05). The prevalence of large for gestational age infants (LGA) was similar in both groups, but birth length of LGA newborns was greater in PCOS women than controls (P<0.05). CONCLUSIONS: PCOS mothers showed a significantly higher prevalence of SGA newborns which cannot be completely attributed to pregnancy complications, and seems to be more related to the PCOS condition of the mother. PMID- 15802313 TI - Associates of self-reported fertility status and infertility treatment-seeking in a rural district of Malawi. AB - BACKGROUND: This study examines the demographic, fertility preference, health/infection and behavioural factors associated with self-reported fertility problems and infertility treatment-seeking in a rural district of Malawi. METHODS: Data come from a population-based survey of 678 women and 362 men. RESULTS: Having a higher ideal number of children than actual number of children, i.e. a 'child deficit', is highly associated with women's reported fertility impairment and treatment-seeking. Other factors associated with women's infertility treatment-seeking are being educated and reporting infertility in self or spouse. In contrast, being in a polygamous union, having exchanged money or goods for sex, and having multiple sex partners are significantly associated with men's reported fertility impairment. Significant factors associated with men's infertility treatment-seeking are having no education, having a long waiting time to pregnancy and having a 'child deficit'. CONCLUSIONS: There is a sex difference in which factors are associated with reported fertility impairment. Fertility preference variables are more often significantly related to women's reported fertility impairment, whereas sexual behaviours are more often significantly related to men's reported fertility impairment. PMID- 15802314 TI - Laparoscopic stripping of endometriomas: a randomized trial on different surgical techniques. Part I: clinical results. AB - BACKGROUND: Laparoscopic surgery has become the gold-standard treatment for ovarian endometriomas. The objective of this trial was to evaluate different procedures for the laparoscopic excision of ovarian endometriomas with the stripping technique. METHODS: Forty-eight patients with ovarian endometrioma were enrolled in two consecutive independent randomized trials. Two different techniques were analysed at the initial adhesion site (circular excision and subsequent stripping versus immediate stripping). Two different techniques were analysed at the ovarian hilus (stripping versus coagulation and cutting). Operative time and technical difficulties were prospectively evaluated. Histological analysis was performed in three portions of the cyst wall and the results of the histologic study are reported separately. RESULTS: At the initial part of the stripping procedure, the technique of circular excision and subsequent stripping appeared to be more easily performed than the technique of direct stripping (P < 0.01), although operative times were comparable between the two techniques. At the hilus, the two techniques utilized appeared to be comparable both for easiness of procedure and operating times. CONCLUSIONS: In this prospective, randomized study, different techniques used during the stripping procedure appeared to be comparable in terms of operative times and complications. One technique used at the beginning of the procedure (circular excision followed by stripping) was easier to perform. PMID- 15802315 TI - Reduction of progesterone receptor expression in human cumulus cells at the time of oocyte collection during IVF is associated with good embryo quality. AB - BACKGROUND: It has been reported that the progesterone receptor (PR) level is transiently increased within the follicle by LH stimulation and controls cumulus cells in follicles and oocyte maturation. The purpose of this study was to predict developmental competence of human oocytes during IVF via analysis of PR in cumulus cells surrounding mature oocytes. METHODS: Prior to oocyte retrieval, the follicular diameter was measured and follicular fluid was collected from each mature follicle. Cumulus cells were manually separated from the oocyte-cumulus complex under a microscope. PR and PR mRNA were assessed by immunohistochemistry and real-time quantitative polymerase chain reaction (RT-PCR) measurement in human cumulus cells. RESULTS: Immunoreactive PR-A was mainly localized in the cytoplasm and PR-B was localized in the nuclei. There was no significant relationship between PR expression and follicular diameter, follicular fluid concentration of steroids, or LH. There was no significant relationship between expression of PRs and fertilization or cleavage rate. However, PR expression was lower in the good morphology group (blastomeres > or =7 cells with fragmentation > or =5% on day 3) when compared to the other groups (P<0.05). CONCLUSIONS: These results suggest that follicular LH or steroids do not affect PR expression, and full reduction of total PR expression on cumulus cells at the time of oocyte collection is associated with good morphology in human oocytes. PMID- 15802316 TI - Absence of hepatotoxicity after long-term, low-dose flutamide in hyperandrogenic girls and young women. AB - BACKGROUND: Flutamide is a pure non-steroidal anti-androgen that may be hepatotoxic, when given in high-dose (750 mg/d). Low- to ultralow-doses (250-62.5 mg/day) have been recently explored in patients with Polycystic Ovary Syndrome (PCOS), and these lower doses were found to confer benefit on multiple PCOS markers. There is a need for evidence on the potential hepatotoxicity of low- and ultralow-dose flutamide therapy. METHODS: We assessed circulating levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as markers of hepatotoxicity in a total of 190 hyperandrogenic girls and young women receiving low- or ultralow-dose flutamide because of established (n = 150) or incipient (n = 40) PCOS without obesity. Assessments were performed before start of flutamide, after 3 months, and subsequently at least twice yearly. RESULTS: AST and ALT results were normal at baseline, and they remained so on flutamide treatment, including between 3 months and last assessment, which was after a mean time of 19 months on low- or ultralow-dose flutamide (range 3-54 months). None of the AST or ALT levels at any time during flutamide treatment was > or = 45 U/L. CONCLUSION: We found no evidence for hepatotoxicity in 190 hyperandrogenic girls or young women receiving low- or ultralow-dose flutamide for up to 54 months. These results may represent a first step in a long process whereby the status of low- and ultralow-dose flutamide may gradually evolve from 'absence of evidence on toxicity' towards 'evidence of absence of hepatic toxicity'. Ultralow-dose flutamide may become a key component within future therapies for hyperandrogenic states in girls and young women. PMID- 15802317 TI - Human ovarian tissue from cortex surrounding benign cysts: a model to study ovarian tissue cryopreservation. AB - BACKGROUND: The scarcity of human ovarian tissue is a major problem in developing research on ovarian cryopreservation. We were interested in ovarian cortex surrounding benign ovarian cysts harvested during their requisite operations. METHODS: Ovarian tissue was collected from 25 women (mean age = 27.7 +/- 1.0 SEM) and frozen in serum-free cryoprotective medium. Histological and viability analysis were performed on fresh and frozen-thawed slices of tissue. RESULTS: Dermoid (n = 7), endometriosis (n = 13) and serous (n = 5) cysts were observed. Follicular densities (expressed per mm3) in ovarian cortex surrounding dermoid cysts were higher than in endometriosis and serous cysts for both histological (median of follicular densities: 13.04, 0.31 and 0.89 respectively) and viability analysis (2.93, 0.05 and 0.71 respectively). Freezing-thawing did not result in gross abnormality of follicle population either in number or morphology (80% of follicles preserved a normal pattern). However, a slight decrease of the density of living follicles (expressed per mm2) was reported. CONCLUSIONS: Ovarian cortex surrounding ovarian cysts, especially dermoid cysts, could be considered a source of ovarian tissue for future research. In our study, the cryopreservation procedure resulted in high follicular survival assessed by both histological and viability analysis. Nevertheless, further studies of in vivo and in vitro follicular maturation are needed to strengthen this model. PMID- 15802318 TI - Association of aromatase (CYP 19) gene variation with features of hyperandrogenism in two populations of young women. AB - BACKGROUND: Aromatase catalyses the conversion of androgens to estrogens and thus variation in the aromatase gene could contribute to female syndromes of androgen excess, such as precocious pubarche (PP) and polycystic ovarian syndrome (PCOS). METHODS: Two groups, one case-control containing girls from Barcelona, Spain with PP (n = 186) or healthy controls (n = 71), and the other a population study of young women from Oxford, UK, who volunteered for a study of normal women's health (n = 109), were genotyped at four aromatase gene haplotype-tag single nucleotide polymorphisms (SNP). Clinical features and hormone concentrations relevant to hyperandrogenism were compared across haplotypes or genotypes. RESULTS: Distributions of aromatase haplotypes (P < 0.0001) and aromatase SNP_50 genotype (P = 0.001) were significantly different between PP girls and Spanish controls. The AGGG haplotype was associated with an odds ratio (95% confidence interval) of 0.5 (0.3-0.9) (P = 0.005) for the presence of PP compared to GAGG. In 84 post pubertal PP girls, aromatase haplotype was associated with functional ovarian hyperandrogenism (P < 0.05), independently of insulin sensitivity. In the Oxford population, SNP_50 was associated with variation in PCOS symptom score (P = 0.008) and circulating testosterone concentrations (P = 0.02). CONCLUSIONS: This study suggests that common variation at the aromatase gene (and not just rare loss-of-function mutations) is associated with androgen excess in girls and young women. PMID- 15802319 TI - Longitudinal study of sperm DNA fragmentation as measured by terminal uridine nick end-labelling assay. AB - BACKGROUND: One major limitation in the use of sperm DNA fragmentation as measured by the TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick end labelling (TUNEL) assay is the paucity of solid data on the stability of this parameter. METHODS: The objective of our study was to evaluate variations in the degree of sperm DNA fragmentation, as measured by the TUNEL assay, over a 6 month period. Five donors provided semen samples (total 107) on the average three times per month, and 10 infertility patients provided semen samples every 4 weeks (total 58). RESULTS: The mean percentage of sperm DNA fragmentation for donors was 13.18%, the within-donor standard deviation (SD(W) = 3.79%) was small compared to between-donor (SD(B) = 17.56%). For the group of patients, the mean percentage of sperm DNA fragmentation was 22.44%, with SD(W) of 4.43% within patients and SD(B) of 29.48% between patients. No seasonal rhythm was observed during the study. The intra-class correlation coefficient for all subjects combined was 0.83. Compared to sperm concentration, individual coefficients of variation for sperm DNA fragmentation indicated less variability in four subjects, but were similar in the others. CONCLUSION: This longitudinal study shows that sperm DNA fragmentation is a parameter with good stability (repeatability) over time; it can be taken as a baseline both in healthy fertile men and in patients from infertility couples. PMID- 15802320 TI - Serum anti-Mullerian hormone levels during controlled ovarian hyperstimulation in women with polycystic ovaries with and without hyperandrogenism. AB - BACKGROUND: Anti-Mullerian hormone (AMH) is expressed in pre- and small-antral follicles. High serum levels are found in women with polycystic ovaries (PCO), accordant with their increased content of small follicles. To evaluate the relationship between AMH, folliculogenesis and hyperandrogenism, we compared serum AMH levels between women with PCO with and without hyperandrogenism and normal controls during controlled ovarian hyperstimulation (COH). METHODS: Nineteen women with PCO and hyperandrogenism (group A), 10 women with PCO but no hyperandrogenism (group B) and 23 ovulatory women with normal ovarian morphology (group C, controls) underwent COH with the long protocol. Serum levels of AMH, estradiol, androstenedione and follicular tracking were determined before gonadotropins treatment (day 0) and every 2-4 days up to the day of HCG administration. RESULTS: AMH levels declined gradually throughout COH in the three groups, but remained higher in groups A and B compared with the controls. Significantly higher levels were found in group A compared with group B, despite comparable numbers of small follicles. Multiple regression analysis revealed that both the number of small follicles and serum androgens were correlated to AMH. CONCLUSIONS: Women with PCO have higher serum AMH levels during COH than controls. Hyperandrogenism is associated with an additional increase in AMH. It is conceivable that hyperandrogenism may reflect more severe disruption of folliculogenesis in women with PCO or may affect AMH secretion. PMID- 15802321 TI - Definition and prevalence of subfertility and infertility. AB - A common definition of sub- and infertility is very important for the appropriate management of infertility. Subfertility generally describes any form of reduced fertility with prolonged time of unwanted non-conception. Infertility may be used synonymously with sterility with only sporadically occurring spontaneous pregnancies. The major factor affecting the individual spontaneous pregnancy prospect is the time of unwanted non-conception which determines the grading of subfertility. Most of the pregnancies occur in the first six cycles with intercourse in the fertile phase (80%). After that, serious subfertility must be assumed in every second couple (10%) although--after 12 unsuccessful cycles- untreated live birth rates among them will reach nearly 55% in the next 36 months. Thereafter (48 months), approximately 5% of the couples are definitive infertile with a nearly zero chance of becoming spontaneously pregnant in the future. With age, cumulative probabilities of conception decline because heterogeneity in fecundity increases due to a higher proportion of infertile couples. In truly fertile couples cumulative probabilities of conception are probably age independent. Under appropriate circumstances a basic infertility work-up after six unsuccessful cycles with fertility-focused intercourse will identify couples with significant infertility problems to avoid both infertility under- and over-treatment, regardless of age: Couples with a reasonably good prognosis (e.g. unexplained infertility) may be encouraged to wait because even with treatment they do not have a better chance of conceiving. The others may benefit from an early resort to assisted reproduction treatment. PMID- 15802322 TI - Meiotic anomalies in infertile men with severe spermatogenic defects. AB - BACKGROUND: This study was aimed at evaluating the rate of pairing failure in pachytene spermatocytes of patients presenting either an obstructive (O) or a non obstructive (NO) infertility. METHODS: Forty-one patients and 13 controls underwent testicular biopsy. Among the patients, 19 had an O infertility and 22 a NO infertility. Preparations of all patients and controls were Giemsa-stained, and synaptonemal complexes from nine of these patients and one control were immunostained. RESULTS: In all, 2931 pachytene nuclei were analysed. The mean rate of asynapsed nuclei from the NO group (25.4%) was significantly higher than that of the O group (9.8%). There was no significant difference between the O group and the controls (10.6%). Immunocytochemistry showed that the number of pachytene nuclei decreased from the early to late pachytene sub-stage in all patients. Two NO patients, one azoospermic and one oligozoospermic, had a high percentage of asynapsed nuclei (86 and 91.8% respectively); one of these patients also presented a precocious localized separation of sister chromatids. CONCLUSION: high levels of extended asynapsis could arise from a primary meiotic defect which may be responsible for 9% of the NO male infertilities at our centre. The prevalence of early pachytene substages suggests that the pachytene checkpoint is localized at the mid-pachytene stage in humans. PMID- 15802323 TI - A qualitative study of women's decision-making at the end of IVF treatment. AB - BACKGROUND: The decision not to pursue further in vitro fertilization (IVF) after one or more unsuccessful attempts is an important and often difficult one for couples. Relatively little is known about the woman's perception of this decision making process. The aim of this study was to examine patients' perspectives of decision-making, including circumstances influencing it and satisfaction with the decision-making process. METHODS: Semi-structured interviews were conducted with a purposive sample of 25 women who had decided to end treatment after unsuccessful IVF treatment. Interviews were tape-recorded and transcribed by means of thematic analysis using the open coding technique. RESULTS: Women experienced difficulty in accepting that their infertility would remain unresolved. Many felt that they had started with unrealistic expectations of treatment success and felt vulnerable to the pressures of both the media and society. Although the decision to end treatment was difficult, it offered many women a way out of the emotional distress caused by IVF; however, the process of decision-making created a sense of 'confrontation' for the women in which they had to address issues they had previously avoided. Adoptive parents perceived less societal pressure than those who remained childless. CONCLUSIONS: Efforts to improve the psychological preparation of couples who decide to end IVF treatment should be directed towards examination of the existing system of consultation, which has certain limitations in terms of the quality of communication and the provision of post-treatment support. Further efforts to develop strategies, which facilitate the decision-making process, should be considered. PMID- 15802324 TI - Exposure to persistent organochlorine pollutants and seminal levels of markers of epididymal and accessory sex gland functions in Swedish men. AB - BACKGROUND: A major exposure route for persistent organochlorine pollutants (POPs) in Sweden is through consumption of fatty fish from the Baltic Sea. Endocrine disruptors, such as POPs, may have a negative impact on sperm quality. The present study aimed to investigate whether exposure to 2,2',4,4',5,5' hexachlorobiphenyl (CB-153) and 1,1-dichloro-2,2-bis (p-chlorophenyl)-ethylene (p,p'-DDE) affects epididymal and accessory sex gland function. METHODS: 157 fishermen from the coastal stretches of Sweden, aged 27-67 years, provided semen samples which were analyzed for prostate-specific antigen (PSA), neutral alpha glucosidase (NAG), fructose and zinc levels. Serum levels of CB-153 and p'p-DDE were determined. RESULTS: The median CB-153 serum level was 189 ng/g lipid (range 40-1460) and a median p,p'-DDE serum level 231 ng/g lipid (range 40-2252). There was a significant linear association between CB-153 and total amount of PSA (slope [beta] = -2.5, 95% confidence interval [CI] -4.0, -0.9; P = 0.02). With age, abstinence time and smoking included in the model the association became non significant (beta = -1.4, 95% CI-3.0, 0.1; P = 0.07). There were no significant associations between CB-153 and zinc, fructose and NAG. As for the exposure variable p,p'-DDE and the outcome variables, no significant associations were found. CONCLUSIONS: The study gives only very limited support of an association between CB-153 in serum and total PSA, and a random finding cannot be excluded. PMID- 15802325 TI - Serum anti-Mullerian hormone levels remain high until late reproductive age and decrease during metformin therapy in women with polycystic ovary syndrome. AB - BACKGROUND: Anti-Mullerian hormone (AMH) is secreted by granulosa cells of ovarian early developing follicles and its serum levels have been shown to correlate with small antral follicle number. Since the pronounced androgen secretion from follicles/stroma in women with polycystic ovary syndrome (PCOS) remains until late reproductive age, and since AMH reflects the number of antral follicles, it was of interest to study the possible age-related relationship between AMH, androgens and follicle number in women with PCOS and in control women. Moreover, the possible effect of metformin on serum AMH levels and the relationship to follicle count and volume were studied. METHODS: Forty-four healthy women (aged 21-44 years) and 65 women with previously diagnosed PCOS (aged 16-44 years) participated in the study. Serum basal AMH levels were correlated with those of serum androstenedione, testosterone, estradiol (E2), LH, FSH and inhibin B, and with follicle number. The effect of metformin on serum AMH concentrations, follicle number and ovarian volume was studied in 26 women (aged 20-41 years) with PCOS after 6 months of treatment. RESULTS: Serum AMH levels were 2- to 3- fold higher in PCOS women than in healthy women. In control women, serum AMH levels correlated positively with those of serum androstenedione (r = 0.564, P < 0.001) and testosterone (r = 0.328, P = 0.036) and negatively with serum FSH concentrations (r = -0.374, P = 0.012) and age (r = -0.691, P<0.001). In women with PCOS, serum AMH levels correlated positively with those of androstenedione (r = 0.311, P = 0.011) and testosterone (r = 0.310, P = 0.011) and with follicle count (r = 0.352, P = 0.012), and negatively with age (r = 0.300, P = 0.014). Serum AMH levels, the number of antral follicles and ovarian volume decreased significantly during metfromin treatment. CONCLUSIONS: Serum AMH levels decreased with age both in healthy women and in women with PCOS, although they were always 2- to 3-fold higher and remained elevated until 40 years of age in PCOS subjects. Thus, since serum AMH levels correlate well with antral follicle count and serum androgen levels, the measurement of AMH could be used as a tool to assess ovarian ageing, to diagnose polycystic ovaries/PCOS and to evaluate treatment efficacy. PMID- 15802326 TI - Comparison of hyaluronate/carboxymethylcellulose membrane and melatonin for prevention of adhesion formation in a rat model. AB - BACKGROUND: To investigate the effectiveness of melatonin in preventing post operative adhesion formation and to compare it with the efficacy of hyaluronate/carboxymethylcellulose membrane in a rat model. MATERIALS AND METHODS: Following pilot studies, 35 rats were operated on in the full study. In 15 animals (group one), 10 standard lesions were inflicted in each uterine horn (total 30 horns) and melatonin was applied before closure of the abdomen. In the second group, 20 animals were operated on and one of the uterine horns (total 20 horns) with standard lesions was treated with hyaluronate/carboxymethylcellulose membrane and the other uterine horn served as a control. Second-look operations were performed 1 week later and adhesion scores were compared. RESULTS: The adhesion scores of uterine horns treated with melatonin and of uterine horns treated with hyaluronate/carboxymethylcellulose membrane were significantly lower than the scores of the controls (P < 0.001). There was no statistically significant difference between the adhesion scores of uterine horns treated with melatonin and of uterine horns treated with hyaluronate/carboxymethylcellulose membrane (P > 0.05). CONCLUSIONS: Both melatonin and hyaluronate/carboxymethylcellulose membrane were effective in prevention of adhesion formation in a rat uterine horn model. PMID- 15802327 TI - Level of FEV1 as a predictor of all-cause and cardiovascular mortality: an effect beyond smoking and physical fitness? PMID- 15802328 TI - Can inhaled steroids mend a broken heart in chronic obstructive pulmonary disease? PMID- 15802329 TI - Th1/Th2 paradigm: not seeing the forest for the trees? PMID- 15802330 TI - Relationship between pulmonary function and lung cancer surgical outcome. AB - The British Thoracic Society and American College of Chest Physician guidelines outline criteria for investigating patients for lung cancer surgery. However, the guidelines are based on relatively old studies. Therefore, the relationship between pulmonary function test results and surgical outcome were studied prospectively in a large cohort of lung cancer patients. From January 2001 to December 2003, 110 patients underwent surgery for lung cancer. All underwent full lung function testing in order to predict post-operative lung function. The hospital mortality rate was 3% and major complication rate 22%. There was poor overall outcome in 13%. Mean pre-operative lung function values were: forced expiratory volume in one second (FEV1) 2.0 L (79.4% of the predicted value), and carbon monoxide diffusing capacity of the lung (D(L,CO)) 73.6% pred. The mean post-operative lung function values were: FEV1 1.4 L (55.6% pred), and D(L,CO) 51.3% pred. All lung function values were better predictors of poor surgical outcome when expressed as a percentage of the predicted value. Using a threshold of pre-operative FEV1 of 47% pred resulted in the most useful positive and negative predictive probabilities, 0.90 and 0.67, respectively. Lung function values expressed as a percentage of the predicted value are more useful predictors of post-operative outcome than absolute values. The threshold of predicted forced expiratory volume in one second for surgical intervention could be lower (45-50% pred) than is currently accepted without increased mortality. PMID- 15802331 TI - Vascular endothelial growth factor in pleural effusions of different origin. AB - This study aimed to determine the diagnostic relevance of vascular endothelial growth factor (VEGF) in the pleural fluid and serum of patients with pleural effusions of different aetiology. VEGF was quantified in the pleural effusion fluid and serum of 96 patients with malignancies (58 lung cancers (CA) and 38 tumours with secondaries to the lung (TM)), 45 with congestive heart failure (CHF), 28 with tuberculosis (TB), 45 with acute infections (INF), and in the serum of 20 healthy controls. VEGF pleural effusion concentrations were significantly different in the main diagnostic groups. VEGF was higher in effusions of patients with malignancies (CA as well as TM) in comparison with INF, TB or CHF. In serum, however, high VEGF concentrations indicated CA, TM or INF, but not TB or CHF. Despite significant differences of VEGF levels in different patient groups, receiver-operating characteristic analysis revealed insufficient diagnostic value of VEGF for differential diagnosis of pleural effusions. In conclusion, vascular endothelial growth factor serum concentration is highly suggestive of the presence of lung disease in general, except for tuberculosis. In effusion fluid, the presence of vascular endothelial growth factor clearly indicates inflammatory or malignant origin. However, for diagnostic use, additional parameters besides vascular endothelial growth factor are mandatory. PMID- 15802332 TI - Interleukin-16 in tuberculous and malignant pleural effusions. AB - The aim of this study was to explore the presence of interleukin (IL)-16 in pleural effusions, the correlation between IL-16 levels and cytological parameters, as well as the chemoattractant activity of IL-16 on CD4+ T lymphocytes. Total nucleated cell and differential counts, and IL-16 concentrations in the pleural effusion from 32 patients with tuberculous pleurisy and 30 patients with lung cancer were determined. Three-colour flow cytometry was performed to determine T-lymphocyte subsets in cell pellets of pleural effusion. The chemoattractant activity of IL-16 for CD4+ T-lymphocytes was also analysed. The levels of IL-16 were significantly higher in tuberculous than in malignant effusions. However, IL-16 levels could not be used for diagnostic purposes due to significant overlap between the two groups. Positive correlations were found between the IL-16 levels and CD4+ T-cells, and pleural fluid was chemotactic for CD4+ T-cells in vitro. Intrapleural administration of IL-16 to patients produced a marked progressive influx of CD4+ T-cells into the pleural space. Compared with malignant pleural effusion, interleukin-16 appeared to be increased in tuberculous pleural effusion. Interleukin-16 levels were positively related to the numbers of CD4+ T-cells, and interleukin-16 could directly induce CD4+ T-cell infiltration into the pleural space. PMID- 15802333 TI - Endothelial interactions of neutrophils under flow in chronic obstructive pulmonary disease. AB - It is generally accepted that the neutrophil is central to the pathogenesis of chronic obstructive pulmonary disease (COPD). Enhanced endothelial interactions of this cell may contribute to the susceptibility of smokers who develop the disease; however, these interactions have not previously been studied in COPD. The aim of the current study was to determine whether enhanced endothelial interactions of neutrophils from smokers are a predisposing factor for the development of COPD. Endothelial interactions under flow and adhesion molecule expression of peripheral blood neutrophils were compared between seven never smokers (NS), seven healthy smokers (HS), 11 COPD patients with severe alpha1 antitrypsin deficiency (PiZ) and neutrophils from 11 COPD patients without the deficiency (PiM). Total adhesive and migratory responses (per mm2 endothelium per 10(6) neutrophils) were significantly greater in the PiM group (mean+/-se 704.2+/ 57.9 versus 509.3+/-48.8 in the PiZ group, 499.3+/-40.1 in the HS and 491.2+/ 33.7 in the NS). This corresponded with increased macrophage antigen-1 (CD11b) expression on stimulated neutrophils in the PiM group compared with the PiZ group (mean+/-se relative fluorescence intensity 1.4+/-0.1 versus 1.1+/-0.1). In conclusion, the enhanced endothelial interaction of neutrophils from smokers who have developed chronic obstructive pulmonary disease in the presence of normal levels of alpha1-antitrypsin deficiency, but not in those with severe alpha(1) antitrypsin deficiency, suggests that this is a predisposing factor for the development of the disease, and upregulation of macrophage antigen-1 may be responsible. PMID- 15802334 TI - Lung function, smoking and mortality in a 26-year follow-up of healthy middle aged males. AB - Lung function has been associated with mortality after adjusting for other risk factors; however, few studies have adjusted for physical fitness and reported separate analyses according to smoking status. In 1972-1975, spirometry, clinical and physiological parameters were recorded in 1,623 apparently healthy males aged 40-59 yrs. After 26 yrs of follow-up, the current authors investigated the association between baseline lung function and mortality, adjusting for smoking, physical fitness and other potential factors. By 2000, 615 individuals (38%) had died, with 308 (50%) of these deaths from cardiovascular (CV) causes. Forced expiratory volume in one second was a predictor of all-cause mortality (risk ratio (RR) 1.10 per reduction of 10%) after adjusting for smoking, physical fitness, age, systolic blood pressure, body mass index and serum cholesterol. The corresponding multivariate RR was 1.07 for CV causes and 1.34 for respiratory death. In conclusion, in stratified analyses among current and former smokers, forced expiratory volume in one second % predicted was a strong independent predictor of all-cause mortality and respiratory death among current smokers. Forced expiratory volume in one second % predicted was not associated with mortality among never-smokers. PMID- 15802335 TI - Decrease of vascular endothelial growth factor in macrophages from long-term smokers. AB - Vascular endothelial growth factor (VEGF), a survival factor for endothelial cells and a promoter of angiogenesis, is reportedly expressed in alveolar macrophages (AMs). To investigate whether long-term smoking with age affects VEGF expression in AMs, bronchoalveolar lavage (BAL) was performed on 18 young and 23 older volunteers with various smoking histories. The expressions of VEGF and its functional receptor, fms-like tyrosine kinase (Flt)-1, were quantified in AMs by real-time RT-PCR and, further, the level of VEGF in BAL fluid was determined by ELISA. VEGF mRNA in AMs demonstrated a 1.8-fold reduction in current smokers compared with nonsmokers in older subjects and, furthermore, a 1.5-fold downregulation in those with emphysema, although there was no difference between current smokers and nonsmokers among the young subjects. The downregulation in total VEGF mRNA was supported by the substantial reduction of VEGF121 and VEGF165 isoforms. However, in contrast, Flt-1 mRNA did not differ within the older groups, whereas it was upregulated in young current smokers compared with age matched nonsmokers. VEGF in BAL fluid is significantly decreased in current smokers compared with nonsmokers, regardless of their age. In conclusion, these data imply that the biological availability of vascular endothelial growth factor in alveolar macrophages is impaired in older current smokers with long-term smoking histories. PMID- 15802336 TI - Low-dose inhaled corticosteroids and the risk of acute myocardial infarction in COPD. AB - Inflammation plays a major role in the development and complications of atherosclerosis. Here, the dose-related impact of inhaled corticosteroids (ICS), used for their anti-inflammatory properties, on the risk of acute myocardial infarction (AMI) is studied in a cohort of chronic obstructive pulmonary disease (COPD) patients. Saskatchewan (Canada) health services databases were used to form a population-based cohort of 5,648 patients, > or =55 yrs, who received a first treatment for COPD between 1990 and 1997. A nested case-control analysis was conducted, where 371 cases presenting with a first AMI were matched with 1,864 controls, based on the date of cohort entry and age. A conditional logistic regression was used to estimate the effect of ICS, after adjusting for use of oral corticosteroids, severity of COPD, sex, systemic hypertension, diabetes and cardiovascular disease. ICS were used in the prior year by 42.2% of cases and 46.4% of controls. Overall, current use of ICS was not associated with a significant decrease in the risk of AMI. However, a 32% reduction in the risk of AMI was observed for doses ranging 50-200 microg x day(-1). In conclusion, very low doses of inhaled corticosteroids may be associated with a reduction in the risk of acute myocardial infarction. PMID- 15802337 TI - Airway inflammation during stable and acutely exacerbated chronic obstructive pulmonary disease. AB - The aim of this study was to clarify the mechanism of increased airway inflammation during an acute exacerbation. A total of 68 chronic obstructive pulmonary disease patients in a stable phase were enrolled and followed-up for 2 3 yrs. Inflammatory cells were analysed, and interleukin (IL)-8, neutrophil elastase, eotaxin, tryptase and RANTES (regulated on activation, normal T-cell expressed and secreted) were measured in sputum, both in a stable phase and during acute exacerbation. Out of 68 patients, 30 (unstable group) developed an acute exacerbation and expectorated adequate sputum during exacerbation. Thirty two patients (stable group) did not develop any exacerbation for 2-3 yrs. The number of neutrophils, lymphocytes and eosinophils, and the levels of IL-8, eosinophil cationic protein (ECP), eotaxin and tryptase in sputum obtained from patients in both groups during the stable phase were significantly higher than those from healthy nonsmokers. There were no significant differences in cell analysis and biomarkers between the two groups, but patients in the unstable group showed more severe airflow limitation. In the unstable group, total cells, lymphocytes, neutrophils and eosinophils, and IL-8, neutrophil elastase, ECP and RANTES levels were significantly increased during an exacerbation from values in a stable phase. These findings suggest that exacerbation of chronic obstructive pulmonary disease may associate with additional increases in airway inflammation mediated by neutrophils, lymphocytes, eosinophils, interleukin-8 and RANTES. PMID- 15802338 TI - Phenotypic characterisation of alveolar macrophages and peripheral blood monocytes in COPD. AB - Alveolar macrophages (AM) participate actively in the inflammatory response that characterises chronic obstructive pulmonary disease (COPD). The present study investigated potential changes in AM phenotypes in patients with COPD. Using flow cytometry, the surface expression of receptors implicated in phagocytosis (CD44, CD36, CD51, CD61, CD14), antigen-presenting capacity (human leukocyte antigen (HLA)-DR), costimulatory molecules (CD80, CD86, CD40) and complement receptor type 3 were assessed in AM from 18 patients with COPD, 14 smokers with normal lung function and nine nonsmokers. When compared to smokers with normal lung function and nonsmokers, the surface expression of HLA-DR and CD80 was lower in AM of patients with COPD. In addition, these patients had a higher percentage of AM with a low level surface expression of CD44. There did not appear to be any difference in the other receptors studied in AM between the three groups. The expression of all these receptors in peripheral blood monocytes also did not differ between groups. In conclusion, these observations suggest that the cell mediated immune function of alveolar macrophages can be reduced in chronic obstructive pulmonary disease, and that this is a local rather than a systemic event. PMID- 15802339 TI - Adoptive transfer of T-helper cell type 1 clones attenuates an asthmatic phenotype in mice. AB - T-helper cell type 1 (Th1) cells have been postulated to have a significant role in protective immunity against allergic diseases. However, recent studies using polarised Th1 cells showed conflicting effects on both airway responsiveness and eosinophilic inflammation in a mouse asthma model. The current study explored the effects of adoptive transfer of established Th1 clones on a murine model of atopic asthma. Mice (BALB/c) were sensitised with ovalbumin (OVA) and challenged with aerosolised OVA (5%, 20 min) for 5 days. Just before starting the first challenge, Th1 clones (5x10(6) x body(-1)) or PBS alone were injected via the tail vein. After assessment of airway responsiveness to methacholine, bronchoalveolar lavage fluid (BALF) was obtained. Histological examination, including morphometric analysis, measurement of cytokines in the BALF and Northern blotting of lung chemokines, was also performed. Adoptive transfer of Th1 clones showed a significantly increased total number of cells, whereas significantly decreased eosinophils were found in the BALF, when compared with mice with injection of vehicle alone or splenic mononuclear cells. Administration of Th1 clones significantly decreased the infiltration of eosinophils but increased mononuclear cells in the peribronchial area. Goblet cell hyperplasia and peribronchial fibrosis were also suppressed by Th1 clones. The transfer of Th1 cells significantly decreased airway responsiveness. Th1 injection significantly increased interferon gamma in the BALF, but significantly decreased interleukin (IL)-5 and IL-13. Eotaxin mRNA was predominantly expressed in the lungs of asthma model mice, whereas RANTES (regulated on activation, normal T cell expressed and secreted) predominates in such mice with Th1 transfer. In conclusion, results suggest that the adoptive transfer of T-helper cell type 1 clones can suppress both lung eosinophilia and airway responsiveness, but increase noneosinophilic inflammation in a mouse model of asthma. PMID- 15802340 TI - The PREVASC study: the clinical effect of a multifaceted educational intervention to prevent childhood asthma. AB - As asthma is the most common chronic disease in childhood, much attention is directed towards primary prevention. Here, the clinical effectiveness of a multifaceted educational prevention was studied. A total of 476 high-risk children were recruited during the prenatal period by general practitioners and randomised to either: 1) a control group, receiving usual care; or 2) an intervention group in which families received instruction from nurses on how to reduce exposure of newborns to mite, pet and food allergens, and passive smoking. A total of 443 infants were followed-up for 2 yrs. At 2 yrs of age, the intervention group (n = 222) had less asthma-like symptoms, including wheezing, shortness of breath and night-time cough, than the control group (n = 221). No significant differences in total and specific immunoglobulin E were found between the groups. During the first 2 yrs of life, the incidence of asthma-like symptoms was similar in both groups; however, subanalysis revealed a significant reduction in the female, but not in the male, intervention group. In conclusion, the intervention used in this study was not effective in reducing asthma-like symptoms in high-risk children during the first 2 yrs of life, although it was modestly effective at 2 yrs. Follow-up is necessary to confirm whether the intervention can actually prevent the development of asthma. PMID- 15802341 TI - Analysis of the cues used by patients when making assessments of their asthma severity. AB - This study measured the relative contribution of four cues: symptoms, effect on activities, emotions and social life, to patients' judgments of overall asthma severity. Judgement analysis techniques were applied to patients' assessments of overall asthma severity using asthma states described in scenarios based on these cues. Altogether, 40 patients, mean age 52 yrs, mean forced expiratory volume in one second 67% predicted, were studied. The relative importance of the cues varied widely between patients; symptoms contributed to 47% of the judgment of asthma severity (range 15-86%), activities 19% (1-73%), emotions 14% (0-43%), and social life 19% (2-57%). Some patients had difficulty with the judgment task, they tended to be older or have worse health. A total of 21 patients had consistent judgment policies. Cluster analysis identified two policy types. In one, symptoms were weighted heavily, in the other all cues were weighted similarly. Policies were not related to disease or demographic factors. Symptoms are important cues for patients when assessing asthma severity, but other cues may be more important in some patients. Patients fall into two groups: those who rely on symptoms and those who use several cues. PMID- 15802342 TI - QT interval dispersion in obstructive sleep apnoea syndrome patients without hypertension. AB - QT interval dispersion (QT(d)) reflects inhomogeneity of repolarisation. Delayed cardiac repolarisation leading to the prolongation of the QT interval is a well characterised precursor of arrhythmias. Obstructive sleep apnoea syndrome (OSAS) can cause cardiovascular complications, such as arrhythmias, myocardial infarction, and systemic and pulmonary hypertension. The aim of this study was to assess QT(d) in OSAS patients without hypertension. A total of 49 subjects without hypertension, diabetes mellitus, any cardiac or pulmonary diseases, or any hormonal, hepatic, renal or electrolyte disorders were referred for evaluation of OSAS. An overnight polysomnography and a standard 12-lead ECG were performed in each subject. According to the apnoea-hypopnoea index (AHI), subjects were divided into control subjects (AHI <5, n = 20) and moderate-severe OSAS patients (AHI > or =15, n = 29). QT(d) (defined as the difference between the maximum and minimum QT interval) and QT-corrected interval dispersion (QT(cd)) were calculated using Bazzet's formula. In conclusion, the QT(cd) was significantly higher in OSAS patients (56.1+/-9.3 ms) than in controls (36.3+/ 4.5 ms). A strong positive correlation was shown between QT(cd) and AHI. In addition, a significantly positive correlation was shown between QT(cd) and the desaturation index (DI). The AHI and DI were significantly related to QT(cd) as an independent variable using stepwise regression analysis. The QT-corrected interval dispersion is increased in obstructive sleep apnoea syndrome patients without hypertension, and it may reflect obstructive sleep apnoea syndrome severity. PMID- 15802343 TI - Resting energy expenditure in Duchenne patients using home mechanical ventilation. AB - Nutritional status is both important and difficult to assess in patients with Duchenne muscular dystrophy (DMD), particularly in those requiring mechanical ventilation (MV). The current authors evaluated body composition (bio impedancemetry), resting energy expenditure (REE; indirect calorimetry) and energy intake in 20 adult patients with DMD using home MV (nocturnal: n = 13; continuous: n = 7) and 12 age-matched healthy controls. The patients were smaller in height than the controls and had a lower body weight. Most of the reduction in body mass index was accounted for by a reduction in fat free mass (FFM). REE (kJ) was significantly reduced in the patients (4559+/-853 kJ x 24 h(-1) versus 7407+/ 1312 kJ x 24 h(-1)), but the difference disappeared after correction for FFM. REE and FFM were correlated in both the controls and patients, but less strongly in the latter, the lower strength of the association being due to the patients using continuous MV (REE and FFM uncorrelated). The food intake of the patients was 1.2+/-0.4 greater than their REE. This study shows that patients with advanced forms of Duchenne muscular dystrophy have balanced energy intakes and resting energy expenditure. PMID- 15802344 TI - Procalcitonin and C-reactive protein levels in HIV-positive subjects with tuberculosis and pneumonia. AB - Pulmonary tuberculosis (PTB) and pneumococcal community-acquired pneumonia (PCAP) are common causes of lower respiratory tract infections in HIV-seropositive patients and may have similar clinical and radiological features. This study aimed to assess the value of serum procalcitonin (PCT) and C-reactive protein (CRP) levels in HIV-seropositive patients with pneumonia, and to investigate their potential role in differentiating pneumococcal from mycobacterial infections. HIV-seropositive patients admitted with pneumonia were evaluated prospectively, 34 with PTB and 33 with PCAP. All 33 patients in the PCAP group and 20 of 34 patients in the PTB group had elevated PCT levels (>0.1 ng x mL( 1)). All patients in both groups had elevated CRP levels (>10 mg x L(-1)). The PTB group had significantly lower CD4 T-lymphocyte counts, lower CRP levels, lower white cell counts, and lower PCT levels than the PCAP group. Receiver operating characteristic analysis showed that optimal discrimination between PTB and PCAP could be performed at a cut-off point of 3 ng x mL(-1) for PCT (sensitivity 81.8%; specificity 82.35%) and 246 mg x L(-1) for CRP (sensitivity 78.8%; specificity 82.3%). In conclusion, HIV-seropositive patients with pneumococcal community-acquired pneumonia had significantly higher procalcitonin and C-reactive protein levels than those with pulmonary tuberculosis. A procalcitonin level >3 ng x mL(-1) and a C-reactive protein level >246 mg x L(-1) were both highly predictive of pneumococcal infection. PMID- 15802345 TI - Relationship between the inoculum dose of Streptococcus pneumoniae and pneumonia onset in a rabbit model. AB - It is generally assumed that the development of bacterial pneumonia becomes possible when the dose of inhaled or aspirated pathogens overwhelms the respiratory tract host defence system, but this hypothesis has not yet been tested either clinically or experimentally. This study evaluated inoculum dose in relation to onset of experimental pneumococcal pneumonia, and estimated the median effective dose resulting in pneumonia in healthy New Zealand White rabbits (mean+/-sd 4.75+/-0.25 kg (n = 27)). Rabbits were endobronchially inoculated with increasing doses of Streptococcus pneumoniae and pneumonia onset was observed over the following 96 h. The diagnostic approach was based on the Clinical Pulmonary Infection Score, modified for use in rabbits. Inoculation of S. pneumoniae at doses of >4.60 log(10) cfu made the development of pneumonia in rabbits more predictable (up to 90%). Lower doses of bacteria failed to cause pneumonia in 80% of inoculated animals. The median effective dose was estimated by means of logistical regression, probit analyses and the Reed-Muench method, and corresponded to 4.32, 4.38 and 4.67 log(10) cfu, respectively. It is speculated that development of pneumococcal pneumonia becomes more likely when the inoculum dose exceeds a threshold of antibacterial protection, making inoculum dose a risk factor for disease onset. PMID- 15802346 TI - CCR5 expression and CC chemokine levels in idiopathic pulmonary fibrosis. AB - CC chemokines play an important role in the pathogenetic mechanisms of interstitial lung disease, while a downregulation of CC chemokine receptor (CCR)5 in the fibrotic stages of sarcoidosis has been observed. To evaluate the involvement of CC chemokines and the expression of CCR5 in idiopathic pulmonary fibrosis (IPF) and, more specifically, in usual interstitial pneumonia, 35 subjects were studied. CC chemokine ligand (CCL)2, CCL3 and CCL4 levels were measured in the bronchoalveolar lavage fluid (BALF) of 18 nonsmoker control subjects and 17 patients affected by IPF. CCR5 expression was evaluated in alveolar macrophages and lymphocytes. The BALF levels of all chemokines were significantly increased in IPF: median (range) CCL3 1.6 (1.0-11.1) versus 1.2 (0.0-3.8) pg x mL(-1); CCL4 6.2 (1.3-96.0) versus 3.4 (0.3-6.8) pg x mL(-1); and CCL2 60.1 (16.7-251.3) versus 4.6 (0.5-119.4) pg x mL(-1). CCL2 levels correlated negatively with the carbon monoxide diffusing capacity of the lung (D(L,CO)) and arterial oxygen tension. CCR5 expression was significantly reduced in lymphocytes from IPF compared with controls. The CC chemokines investigated are involved in the inflammatory mechanisms of idiopathic pulmonary fibrosis, and the results are in agreement with the hypothesis of a downregulation of the T-helper 1 immunological response in this disease. PMID- 15802347 TI - Angiotensin II antagonism fails to ameliorate bleomycin-induced pulmonary fibrosis in mice. AB - Based on current evidence, transforming growth factor (TGF)-beta plays a central pathogenic role in the development of pulmonary fibrosis. There is growing evidence that angiotensin II can serve as a stimulus for TGF-beta-mediated lung fibrosis. However, the role of angiotensin II in the pathobiology of pulmonary fibrosis in vivo remains unclear and the therapeutic potential for targeting angiotensin II in a bleomycin-induced pulmonary fibrosis model is not well known. Therefore, the aim of this study was to test whether the angiotensin II antagonist, losartan, attenuated the development of bleomycin-induced pulmonary fibrosis in two distinct murine strains, C57/BL6 and Sv129. This was determined by histopathology and quantification of collagen content by hydroxyproline assay. Despite demonstrable angiotensin II antagonism in vivo and a reduction in measures of acute lung injury, losartan therapy, at a dose shown to reduce renal and cardiac fibrosis in mice, failed to significantly ameliorate bleomycin induced pulmonary fibrosis. In conclusion, these data suggest that the pulmonary fibrotic disease process in vivo is not solely dependent on angiotensin II activity and the potential for angiotensin II receptor blockers as a therapeutic strategy in patients with pulmonary fibrosis may be limited. PMID- 15802348 TI - New criteria for impaired fasting glucose and screening for diabetes in cystic fibrosis. AB - Cystic fibrosis-related diabetes mellitus (CFRD) is the most frequent comorbidity in cystic fibrosis. Its clinical relevance is stressed by the association with increased mortality, and decreased pulmonary and nutritional status. An annual oral glucose tolerance test (OGTT) is recommended as a screening test for CFRD, but this is often not realised because of its time- and resource-consuming nature. Therefore, alternative approaches are welcome. In 2003, the American Diabetes Association (ADA) lowered the cut-off point separating normal from elevated fasting plasma glucose from <6.1 mmol x L(-1) to <5.6 mmol x L(-1), suggesting the performance of an OGTT only in those with impaired fasting glucose (IFG; range 5.6-6.0 mmol x L(-1)). The current authors tested whether this approach was reliable for the early identification of patients with CFRD. OGTTs from 1,128 patients (53% males; 47% females; median age 17.1 yrs) were available for analysis. A total of 101 (8.9%) OGTTs were classified as diabetic. The new ADA criteria for IFG increased the sensitivity to 82% (versus 65%) and decreased the specificity to 70% (versus 94%) compared with the old criteria used to identify patients with diabetic OGTTs. In conclusion, the American Diabetes Association approach of using impaired fasting glucose as an indication for performing selective oral glucose tolerance tests is definitely unsuitable when aiming at the early identification of patients with cystic fibrosis-related diabetes mellitus, and it cannot replace annual oral glucose tolerance tests. PMID- 15802349 TI - Lung function in adults with cystic fibrosis at altitude: impact on air travel. AB - Current guidelines for air travel state that patients with chronic respiratory diseases are required to use oxygen if their in-flight arterial oxygen tensions (Pa,O2) drop below 6.6 kPa. This recommendation may not be strictly applicable to cystic fibrosis patients, who may tolerate lower Pa,O2 for several hours without clinical symptoms. Lung function, symptoms, blood gas levels and signs of pulmonary hypertension were studied in 36 cystic fibrosis patients at altitudes of 530 m and, after 7 h, 2,650 m. A hypoxia inhalation test (inspiratory oxygen fraction 0.15) was performed at low altitude in order to predict high-altitude hypoxaemia. Median Pa,O2 dropped from 9.8 kPa at low altitude to 7.0 kPa at high altitude. Mild exercise at a workload of 30 W further decreased Pa,O2. Two-thirds of all patients exhibited Pa,O2 of <6.6 kPa during exercise and, except for one patient, were asymptomatic. Patients were significantly less obstructed at an altitude of 2,650 m. Low forced expiratory volume in one second at baseline was associated with a low Pa,O2 at altitude. It is concluded that cystic fibrosis patients with baseline arterial oxygen tensions of >8.0 kPa safely tolerate an altitude of 2,650 m for several hours under resting conditions. The risk assessment of low in-flight oxygenation should encompass the whole clinical situation of cystic fibrosis patients, with special attention being paid to the presence of severe airway obstruction. PMID- 15802350 TI - Hypoxaemia in chronic obstructive pulmonary disease patients during a commercial flight. AB - The aim of the study was to investigate hypoxaemia in chronic obstructive pulmonary disease patients during a commercial flight. The effect of a commercial flight, lasting 5 h 40 min, on arterial blood gas levels and symptoms in 18 chronic obstructive pulmonary disease patients with a pre-flight percutaneous oxygen saturation of 94% and self-reported ability to walk 50 m without severe dyspnoea was studied. The arterial oxygen tension (Pa,O2) decreased from sea level to cruising altitude (10.3+/-1.2 versus 8.6+/-0.8 kPa), but, thereafter, except for one patient, remained stable throughout the flight. During light exercise, however, there was further desaturation (percutaneous oxygen saturation 90+/-4 versus 87+/-4%). After 4 h, a decrease in arterial carbon dioxide tension (5.0+/-0.4 versus 4.8+/-0.4 kPa) and an increase in cardiac frequency (87+/-13 versus 95+/-13 beats x min(-1)) were observed. A pre-flight Pa,O2 of >9.3 kPa did not secure an acceptable in-flight Pa,O2. Aerobic capacity showed the strongest correlation with in-flight Pa,O2. In conclusion, following an initial decrease in arterial oxygen tension, chronic obstructive pulmonary disease patients in a stable state of their disease seem to maintain a stable arterial oxygen tension throughout a flight of intermediate duration, except when walking along the aisle. However, a decrease in arterial carbon dioxide tension, indicating compensatory hyperventilation, could imply a risk of respiratory fatigue during longer flights. PMID- 15802351 TI - Asthma during pregnancy: mechanisms and treatment implications. AB - Asthma is becoming increasingly prevalent worldwide. Numerous historical and prospective cohort studies have investigated the effects of maternal asthma on pregnancy outcome; however, the data has been conflicting and many studies have not used standard classifications for asthma severity. Overall, the literature suggests that asthmatic females are more at risk of low birth weight neonates, pre-term delivery and complications such as pre-eclampsia, especially in the absence of actively managed asthma treated with inhaled corticosteroids. Pregnancy with a female foetus may particularly increase the risk of these outcomes. In addition, pregnancy has an effect on the course of asthma. The risk of an exacerbation requiring medical intervention may be as high as 50% in females with severe asthma and this may further increase the risk of poor outcomes, particularly low birth weight and pre-term delivery. The mechanisms responsible for changes in asthma with pregnancy, or alterations in pregnancy outcomes due to asthma have not been thoroughly explored. Maternal inflammatory pathways may contribute to reduced foetal growth through alterations in placental function. Asthma treatment, by reducing maternal inflammation and preventing exacerbations, is safe for use in pregnant females and contributes to improved outcomes for both mother and foetus. PMID- 15802352 TI - Should tuberculosis treatment and control be addressed differently in HIV infected and -uninfected individuals? AB - Infection with HIV drives the tuberculosis epidemic, especially in sub-Saharan Africa, where up to 75% of individuals with tuberculosis are co-infected with HIV. This article reviews the epidemiological link between the conditions, how tuberculosis diagnosis and treatment differ between HIV-infected versus uninfected individuals and the span of additional measures required to prevent and control HIV-related tuberculosis. Tuberculosis chemotherapy using standard short-course regimens is highly effective in both groups, and treatment follows the same principles. It differs in certain aspects, such as when antiretroviral treatment should be started in HIV-infected individuals with tuberculosis and consideration of drug-drug interactions between the rifamycins and certain antiretroviral drugs. Control of HIV-related tuberculosis requires, fundamentally, control of HIV transmission. Meanwhile, it is necessary to make concentrated efforts to intensify high-quality tuberculosis services employing the directly observed treatment, short-course (DOTS) strategy, carry out extensive research towards an evidence-based model for the expanded scope of collaborative tuberculosis and HIV/AIDS interventions, and ensure efficient implementation of the findings and recommended policies. The challenge is gigantic, and both robust within-country and international leadership and competent management capabilities will be required, in addition to substantial human and financial resources. PMID- 15802353 TI - Noninvasive assessment and monitoring of the pulmonary circulation. AB - In pulmonary vascular disease, changes in the pulmonary vascular bed will lead to altered pulmonary haemodynamics. This review describes the application of several physiological principles to measure these changes noninvasively by means of novel techniques. Flow characteristics of blood through the pulmonary vascular bed alter in pulmonary vascular disease. Recent developments in magnetic resonance imaging and computed tomography make it possible to visualise and quantify these abnormal flow patterns. Information regarding pulmonary perfusion can also be obtained by measuring the electrical impedance changes in the lung by electrical impedance tomography. A more indirect approach to measure the pulmonary blood flow is the measurement of the absorption of acetylene, a perfusion limited gas. Information on the pulmonary vascular bed can also be obtained by the measurement of exhaled products of the pulmonary vascular endothelium, such as nitric oxide. Although all the techniques described offer new ways to diagnose or monitor pulmonary vascular disease, clinical data on these techniques are limited. Further improvement and evaluation of the clinical value of these techniques are therefore obligatory before they can be used in clinical practice. PMID- 15802354 TI - A 75-year-old female with dyspnoea and chest pain. PMID- 15802355 TI - Azithromycin in cystic fibrosis. PMID- 15802356 TI - Flow limitation and dynamic hyperinflation. PMID- 15802357 TI - Value of imprint cytology for ultrasound-guided transthoracic core biopsy. PMID- 15802359 TI - Optimizing management of patients with coronary artery disease: how do we get there? PMID- 15802360 TI - Management and outcome of patients with established coronary artery disease: the Euro Heart Survey on coronary revascularization. AB - AIMS: The purpose of the Euro Heart Survey Programme of the European Society of Cardiology is to evaluate to which extent clinical practice endorses existing guidelines as well as to identify differences in population profiles, patient management, and outcome across Europe. The current survey focuses on the invasive diagnosis and treatment of patients with established coronary artery disease (CAD). METHODS AND RESULTS: Between November 2001 and March 2002, 7769 consecutive patients undergoing invasive evaluation at 130 hospitals (31 countries) were screened for the presence of one or more coronary stenosis >50% in diameter. Patient demographics and comorbidity, clinical presentation, invasive parameters, treatment options, and procedural techniques were prospectively entered in an electronic database (550 variables+29 per diseased coronary segment). Major adverse cardiac events (MACE) were evaluated at 30 days and 1 year. Out of 5619 patients with angiographically proven coronary stenosis (72% of screened population), 53% presented with stable angina while ST elevation myocardial infarction (STEMI) was the indication for coronary angiography in 16% and non-ST segment elevation myocardial infarction or unstable angina in 30%. Only medical therapy was continued in 21%, whereas mechanical revascularization was performed in the remainder [percutaneous coronary intervention (PCI) in 58% and coronary artery bypass grafting (CABG) in 21%]. Patients referred for PCI were younger, were more active, had a lower risk profile, and had less comorbid conditions. CABG was performed mostly in patients with left main lesions (21%), two- (25%), or three-vessel disease (67%) with 4.1 diseased segments, on average. Single-vessel PCI was performed in 82% of patients with either single- (45%), two (33%), or three-vessel disease (21%). Stents were used in 75% of attempted lesions, with a large variation between sites. Direct PCI for STEMI was performed in 410 cases, representing 7% of the entire workload in the participating catheterization laboratories. Time delay was within 90 min in 76% of direct PCI cases. In keeping with the recommendations of practice guidelines, the survey identified under-use of adjunctive medication (GP IIb/IIIa receptor blockers, statins, and angiotensin-converting enzyme-inhibitors). Mortality rates at 30 days and 1 year were low in all subgroups. MACE primarily consisted of repeat PCI (12%). CONCLUSION: The current Euro Heart Survey on coronary revascularization was performed in the era of bare metal stenting and provides a global European picture of the invasive approach to patients with CAD. These data will serve as a benchmark for the future evaluation of the impact of drug-eluting stents on the practice of interventional cardiology and bypass surgery. PMID- 15802361 TI - The challenges of generating evidence to guide mechanical circulatory support based management of advanced heart failure. PMID- 15802362 TI - Reduction of QRS duration following pulmonary valve replacement in tetralogy of Fallot: implications for arrhythmia reduction? PMID- 15802363 TI - The role of carbonic anhydrase in the recovery of skeletal muscle from anoxia. AB - To investigate the role of carbonic anhydrase in the recovery of skeletal muscle from anoxia, pH and cell phosphates were measured by (31)P-nuclear magnetic resonance in superfused newborn rabbit myotubes and cultured mouse soleus cells (H-2K(b)-ts a58) after approximately 2-3.5 h without superfusion. In control studies, pH and phosphocreatine fell and P(i) rose during anoxia and recovered within <10 min after reperfusion began. A carbonic anhydrase inhibitor, acetazolamide, and dimethylamiloride, an inhibitor of the Na(+)/H(+) antiporter NHE1, delayed the recoveries of pH, phosphocreatine, and P(i) for >10 min, but the rate of recovery, once initiated, was unchanged. In the presence of the inhibitors, after reperfusion started, the pH did not rise immediately, despite a large inwardly directed HCO(3)(-) gradient, suggesting that HCO(3)(-) movement was unimportant in acid elimination. Lactate, measured by its methyl protons, rose during anoxia and did not fall after 1 h of reperfusion and could not have eliminated protons by cotransport. We conclude that NHE1 is the major exporter of protons by skeletal muscle in recovery from a period of anoxia and that it is essential for functioning carbonic anhydrase to be attached to NHE1 to activate it. The mechanism of late recovery of pH could be the mobilization of another proton transporter or removal of the inhibition of the Na(+)/H(+) antiporter. Inhibition of carbonic anhydrase in skeletal muscle retards acid removal and modifies muscle metabolism significantly after anoxia. PMID- 15802364 TI - Thromboxane inhibition reduces an early stage of chronic hypoxia-induced pulmonary hypertension in piglets. AB - The pulmonary vasoconstrictor, thromboxane, may contribute to the development of pulmonary hypertension. Our objective was to determine whether a combined thromboxane synthase inhibitor-receptor antagonist, terbogrel, prevents pulmonary hypertension and the development of aberrant pulmonary arterial responses in newborn piglets exposed to 3 days of hypoxia. Piglets were maintained in room air (control) or 11% O(2) (hypoxic) for 3 days. Some hypoxic piglets received terbogrel (10 mg/kg po bid). Pulmonary arterial pressure, pulmonary wedge pressure, and cardiac output were measured in anesthetized animals. A cannulated artery technique was used to measure responses to acetylcholine. Pulmonary vascular resistance for terbogrel-treated hypoxic piglets was almost one-half the value of untreated hypoxic piglets but remained greater than values for control piglets. Dilation to acetylcholine in preconstricted pulmonary arteries was greater for terbogrel-treated hypoxic than for untreated hypoxic piglets, but it was less for pulmonary arteries from both groups of hypoxic piglets than for control piglets. Terbogrel may ameliorate pulmonary artery dysfunction and attenuate the development of chronic hypoxia-induced pulmonary hypertension in newborns. PMID- 15802365 TI - Effect of pulmonary perfusion on the slopes of single-breath test of CO2. AB - The objective of this study was to evaluate the effects of lung perfusion on the slopes of phases II (S(II)) and III (S(III)) of a single-breath test of CO(2) (SBT-CO(2)). Fourteen patients submitted to cardiac surgery were studied during weaning from cardiopulmonary bypass (CPB). Pump flow was decreased in 20% steps, from 100% (total CPB = 2.5 l.min(-1).m(-2)) to 0%. This maneuver resulted in a progressive and opposite increase in pulmonary blood flow (PBF) while maintaining ventilator settings constant. SBT-CO(2), respiratory, and hemodynamic variables remained unchanged before and after CPB, reflecting a constant condition at those stages. S(III) was similar before and after CPB (19.6 +/- 2.8 and 18.7 +/- 2.1 mmHg/l, respectively). S(III) was lowest during 20% PBF (8.6 +/- 1.9 mmHg/l) and increased in proportion to PBF until exit from CPB (15.6 +/- 2.2 mmHg/l; P < 0.05). Similarly, S(II) and the CO(2) area under the curve increased from 163 +/- 41 mmHg/l and 4.7 +/- 0.6 ml, respectively, at 20% PBF to 313 +/- 32 mmHg/l and 7.9 +/- 0.6 ml (P < 0.05) at CPB end. When S(II) and S(III) were normalized by the mean percent expired CO(2), they remained unchanged during the protocol. In summary, the changes in PBF affect the slopes of the SBT-CO(2). Normalizing S(II) and S(III) eliminated the effect of changes in the magnitude of PBF on the shape of the SBT-CO(2) curve. PMID- 15802366 TI - Anatomically based finite element models of the human pulmonary arterial and venous trees including supernumerary vessels. AB - Studies of the origin of pulmonary blood flow heterogeneity have highlighted the significant role of vessel branching structure on flow distribution. To enable more detailed investigation of structure-function relationships in the pulmonary circulation, an anatomically based finite element model of the arterial and venous networks has been developed to more accurately reflect the geometry found in vivo. Geometric models of the arterial and venous tree structures are created using a combination of multidetector row X-ray computed tomography imaging to define around 2,500 vessels from each tree, a volume-filling branching algorithm to generate the remaining accompanying conducting vessels, and an empirically based algorithm to generate the supernumerary vessel geometry. The explicit generation of supernumerary vessels is a unique feature of the computational model. Analysis of branching properties and geometric parameters demonstrates close correlation between the model geometry and anatomical measures of human pulmonary blood vessels. A total of 12 Strahler orders for the arterial system and 10 Strahler orders for the venous system are generated, down to the equivalent level of the terminal bronchioles in the bronchial tree. A simple Poiseuille flow solution, assuming rigid vessels, is obtained within the arterial geometry of the left lung, demonstrating a large amount of heterogeneity in the flow distribution, especially with inclusion of supernumerary vessels. This model has been constructed to accurately represent available morphometric data derived from the complex asymmetric branching structure of the human pulmonary vasculature in a form that will be suitable for application in functional simulations. PMID- 15802367 TI - Impact of aging on muscle blood flow in chronic heart failure. AB - Chronic heart failure (CHF) is manifested principally in the elderly population. Therefore, to understand the causes of exercise intolerance in CHF patients, it is imperative to resolve the effects of aging on muscle blood flow (BF) in CHF. To address this issue, we determined the muscle BF response to submaximal treadmill exercise (20 m/min, 5% grade) in young (Y(CHF): 6-8 mo, 412 +/- 11 g, n = 11) and old (O(CHF): 27-29 mo, 494 +/- 10 g, n = 8) Fischer 344 x Brown Norway rats with similar degrees of myocardial infarction-induced left ventricular (LV) dysfunction [resting LV end-diastolic pressure: Y(CHF) = 24 +/- 2, O(CHF) = 22 +/ 2 mmHg; derivative of LV pressure over time: Y(CHF) = 5,168 +/- 285; O(CHF) = 5,050 +/- 165 mmHg/s; lung weight normalized to body weight: Y(CHF) = 9.14 +/- 0.72; O(CHF) = 8.21 +/- 0.29 mg/g (all P > 0.05)]. The exercising heart rate response was blunted in O(CHF) compared with Y(CHF) rats (Y(CHF) = 454 +/- 8, O(CHF) = 395 +/- 9 beats/min; P < 0.05). BF (radiolabeled microspheres) to the total hindlimb musculature and to each of the 28 individual muscles examined was similar between Y(CHF) and O(CHF) rats under resting conditions. During exercise, BF to five of the hindlimb muscles that normally possess a majority of slow twitch oxidative and fast-twitch oxidative glycolytic muscle fibers increased significantly less (-25 to -42%) for O(CHF) compared with Y(CHF) rats. In contrast, BF to 14 of the hindlimb muscles that normally possess a majority of fast-twitch glycolytic muscle fibers was increased (+22 to +337%) for O(CHF) vs. Y(CHF) rats, which contributed to a greater mass-specific total hindlimb BF response in O(CHF) rats (Y(CHF) = 78 +/- 5, O(CHF) = 100 +/- 11 ml.min(-1).100 g( 1); P < 0.05) and coincided with greater reductions in BF to the kidneys and splanchnic organs during exercise in O(CHF) vs. Y(CHF). In conclusion, there appears to be a profound age-related redistribution of BF from the highly oxidative to the highly glycolytic muscles of the hindlimb during exercise in O(CHF) compared with Y(CHF) rats. This phenomenon is qualitatively similar to that reported previously for healthy young and old rats. PMID- 15802368 TI - Epithelial cell deformation during surfactant-mediated airway reopening: a theoretical model. AB - A theoretical model is presented describing the reopening by an advancing air bubble of an initially liquid-filled collapsed airway lined with deformable epithelial cells. The model integrates descriptions of flow-structure interaction (accounting for nonlinear deformation of the airway wall and viscous resistance of the airway liquid flow), surfactant transport around the bubble tip (incorporating physicochemical parameters appropriate for Infasurf), and cell deformation (due to stretching of the airway wall and airway liquid flows). It is shown how the pressure required to drive a bubble into a flooded airway, peeling apart the wet airway walls, can be reduced substantially by surfactant, although the effectiveness of Infasurf is limited by slow adsorption at high concentrations. The model demonstrates how the addition of surfactant can lead to the spontaneous reopening of a collapsed airway, depending on the degree of initial airway collapse. The effective elastic modulus of the epithelial layer is shown to be a key determinant of the relative magnitude of strains generated by flow-induced shear stresses and by airway wall stretch. The model also shows how epithelial-layer compressibility can mediate strains arising from flow-induced normal stresses and stress gradients. PMID- 15802369 TI - Effect of physical activity on MRI-measured blood oxygen level-dependent transients in skeletal muscle after brief contractions. AB - The signal intensity (SI) in gradient-echo, echo-planar magnetic resonance images (repetition time/echo time = 1,000/40) of anterior tibialis muscle in active [estimated energy expenditure 42.4 +/- 3.7 (SD), n = 8] vs. sedentary (32.3 +/- 0.6 kcal.kg(-1).day(-1), n = 8) young adult (18-34 yr old) human subjects was measured after single, 1-s-duration maximum voluntary ankle dorsiflexion contractions. There was no difference between groups in anterior tibial muscle cross-sectional area or peak force. In both groups there was a transient increase in anterior tibialis muscle SI, which peaked 5-7 s after the end of each contraction. The magnitude of the SI transient was over threefold greater [5.5 +/ 1.0 (SE) vs. 1.5 +/- 0.4%] and persisted twice as long (half-recovery time 5.4 +/- 0.4 vs. 2.7 +/- 0.3 s) in the active subjects. In the same subjects, blood flow in popliteal, anterior tibial, and posterior tibial arteries was measured by cardiac-gated CINE magnetic resonance angiography before and after 2 min of dynamic, repetitive ankle dorsiflexion exercise. There was no difference between groups in resting or postexercise flow in anterior tibial artery, although popliteal and posterior tibial artery flow after exercise tended to be greater in the active group. The results indicate that transient hyperemia and oxygenation in muscle after single contractions are enhanced by chronic physical activity to a greater extent than peak muscle blood flow. PMID- 15802370 TI - Selective alpha2-adrenergic properties of dexmedetomidine over clonidine in the human forearm. AB - We tested the hypothesis that dexmedetomidine (Dex) has greater alpha(2)- vs. alpha(1) selectivity than clonidine and causes more alpha(2)-selective vasoconstriction in the human forearm. After local beta-adrenergic blockade with propranolol, forearm blood flow (plethysmography) responses to brachial artery administration of Dex, clonidine, and phenylephrine (alpha(1)-agonist) were determined in healthy young adults before and after alpha(2)-blockade with yohimbine (n = 10) or alpha(1)-blockade with prazosin (n = 9). Yohimbine had no effect on phenylephrine-mediated vasoconstriction but blunted Dex-mediated vasoconstriction (mean +/- SE: -41 +/- 5 vs. -11 +/- 2%; before vs. after yohimbine) more than clonidine-mediated vasoconstriction (-39 +/- 5 vs. -28 +/- 4%; before vs. after yohimbine) (P < 0.02). Prazosin blunted phenylephrine mediated vasoconstriction (-39 +/- 4 vs. -8 +/- 2%; before vs. after prazosin) but had similar effects on both Dex- (-30 +/- 4 vs. -39 +/- 6%; before vs. after prazosin) and clonidine-mediated vasoconstriction (-29 +/- 3 vs. -41 +/- 7%; before vs. after prazosin) (P > 0.7). Both Dex and clonidine reduced deep forearm venous norepinephrine concentrations to a similar extent (-59 +/- 12 vs. -55 +/- 10 pg/ml; Dex vs. clonidine, P > 0.6); this effect was abolished by yohimbine and blunted by prazosin. These results suggest that Dex causes more alpha(2) selective vasoconstriction in the forearm than clonidine. The similar vasoconstrictor responses to both drugs after prazosin might be explained by the presynaptic effects on norepinephrine release. PMID- 15802371 TI - Acoustic rhinometry in humans: accuracy of nasal passage area estimates, and ability to quantify paranasal sinus volume and ostium size. AB - A comprehensive study that compared acoustic rhinometry (AR) data to computed tomography (CT) data was performed to evaluate the accuracy of AR measurements in estimating nasal passage area and to assess its ability of quantifying paranasal sinus volume and ostium size in live humans. Twenty nasal passages of 10 healthy adults were examined by using AR and CT. Actual cross-sectional areas of the nasal cavity, sinus ostia sizes, and maxillary and frontal sinus volumes were determined from CT sections perpendicular to the curved acoustic axis of the nasal passage. Nasal cavity volume (from nostril to choana) calculated from the AR-derived area-distance curve was compared with that from the CT-derived area distance curve. AR measurements were also done on pipe models that featured a side branch (Helmholtz resonator of constant volume but two different neck diameters) simulating a paranasal sinus. In the anterior nasal cavity, there was good agreement between the cross-sectional areas determined by AR and CT. However, posterior to the sinus ostia, AR overestimated cross-sectional area. The difference between AR nasal volume and CT nasal volume was much smaller than the combined volume of the maxillary and frontal sinuses. The results suggest that AR measurements of the healthy adult nasal cavity are reasonably accurate to the level of the paranasal sinus ostia. Beyond this point, AR overestimates cross sectional area and provides no quantitative data for sinus volume or ostium size. The effects of paranasal sinuses and acoustic resonances in the nasal cavity are not accounted for in the present AR algorithms. PMID- 15802372 TI - The mineralocorticoid receptor: a journey exploring its diversity and specificity of action. AB - The mineralocorticoid receptor (MR) integrates hormonal signaling and activates the expression of aldosterone target genes, which control various physiological processes. In recent years, evidence has been provided for an important role of MR not only in the regulation of sodium and water homeostasis but also in cardiovascular function, neuronal fate, and adipocyte differentiation. MR belongs to the steroid receptor family that displays common mechanism of action. As a result, some apparent similarities with the glucocorticoid receptor (GR) have shaded MR's own specificities. The description of its gene structure, messenger isoforms, protein variants, functional domains, and posttranslational modifications (phosphorylation, ubiquitinylation, sumoylation, acetylation) as well as a panel of interactions with coregulators, progressively depicted an original portrait of MR and shed light on its specific mechanism of action. In this review, after an overview of MR characteristics, the multiple levels of MR selectivity over other steroid receptors, in particular GR, will be described as well as the consequences for aldosterone-regulated gene expression. PMID- 15802373 TI - Pituitary resistance to thyroid hormone syndrome is associated with T3 receptor mutants that selectively impair beta2 isoform function. AB - Resistance to thyroid hormone (RTH) syndrome is an inherited inability to respond appropriately to T3 hormone. In generalized RTH, the T3 response of both the pituitary and periphery is disrupted. In pituitary (or central) RTH, the ability of the pituitary to sense (and down-regulate) elevated T3 is selectively impaired, whereas the periphery remains relatively T3 responsive, resulting in peripheral thyrotoxicity. Both forms of disease are linked to mutations in thyroid hormone receptor (TR)-beta. TRbeta is expressed by alternate mRNA splicing as two isoforms: TRbeta2, found primarily in the pituitary/hypothalamus, and TRbeta1, expressed broadly in many tissues. We report here that the wild-type TRbeta2 isoform displays an enhanced T3 response relative to the TRbeta1 isoform. Mutations associated with generalized RTH (P453S, G345S) impair both TRbeta2 and TRbeta1 function proportionally, whereas mutations associated with pituitary specific RTH (R338L, R338W, R429Q) disproportionately disrupt TRbeta2 function. We propose that in the normal organism, and in generalized RTH, TRbeta2 in the pituitary can sense rising T3 levels in advance of TRbeta1 in the periphery, preventing thyrotoxicity. In contrast, the TRbeta mutations associated with pituitary RTH disproportionately disrupt the pituitary's ability to sense and suppress elevated T3 levels in advance of the periphery, producing symptoms of thyrotoxicity. PMID- 15802374 TI - Processing of pro-islet amyloid polypeptide in the constitutive and regulated secretory pathways of beta cells. AB - Islet amyloid is a pathologic characteristic of the pancreas in type 2 diabetes comprised mainly of the beta-cell peptide islet amyloid polypeptide (IAPP; amylin). We used a pulse-chase approach to investigate the kinetics of processing and secretion of the IAPP precursor, proIAPP, in beta cells. By only 20 min after synthesis, a COOH-terminally processed proIAPP intermediate (approximately 6 kDa) was already present in beta cells. Formation of this NH2-terminally extended intermediate was not prevented by arresting secretory pathway transport at the trans-Golgi network (TGN) by either brefeldin A or temperature blockade, suggesting that this initial cleavage step occurs in the TGN before entry of (pro)IAPP into granules. Mature IAPP (approximately 4 kDa) was not detected until 60 min of chase, suggesting that NH2-terminal cleavage occurs in granules. Cells chased in low glucose without Ca2+ or with diazoxide, to block regulated release, secreted both proIAPP (approximately 8 kDa) and a partially processed form (approximately 6 kDa) via the constitutive secretory pathway. Stimulation of regulated secretion resulted in secretion primarily of mature IAPP as well as low levels of both unprocessed (approximately 8 kDa) and partially processed (approximately 6 kDa) proIAPP. We conclude that normal processing of proIAPP is a two-step process initiated by cleavage at its COOH terminus (likely by prohormone convertase 1/3 in the TGN) followed by cleavage at its NH2 terminus (by prohormone convertase 2 in granules) to form IAPP. Both proIAPP and its NH2 terminally extended intermediate appear to be normal secretory products of the beta cell that can be released via either the regulated or constitutive secretory pathways. PMID- 15802375 TI - Cell cycle progression stimulated by tamoxifen-bound estrogen receptor-alpha and promoter-specific effects in breast cancer cells deficient in N-CoR and SMRT. AB - Estrogen receptor alpha (ERalpha) mediates the effects of estrogens in breast cancer development and growth via transcriptional regulation of target genes. Tamoxifen can antagonize ERalpha activity and has been used in breast cancer therapy. Tamoxifen-bound ERalpha associates with nuclear receptor corepressor (N CoR) and silencing mediator for retinoid and thyroid hormone receptors (SMRT) at certain target genes. Here we show the effects of reducing N-CoR and SMRT levels on the actions of estrogen and tamoxifen in breast cancer cells. Silencing both corepressors led to tamoxifen-stimulated cell cycle progression without activation of the ERalpha target genes c-myc, cyclin D1, or stromal cell-derived factor 1, which play a role in estrogen-induced proliferation. By contrast, expression of X-box binding protein 1 was markedly elevated in tamoxifen-treated cells in which N-CoR and SMRT had been silenced. The gain in cell cycle entry seen with tamoxifen when N-CoR and SMRT were silenced was dependent on ERalpha and not observed upon treatment with estradiol or epidermal growth factor. These results suggest that N-CoR and SMRT play an active role in preventing tamoxifen from stimulating proliferation in breast cancer cells through repression of a subset of target genes involved in ERalpha function and cell proliferation. PMID- 15802376 TI - Estrogen receptor alpha and beta heterodimers exert unique effects on estrogen- and tamoxifen-dependent gene expression in human U2OS osteosarcoma cells. AB - The 17beta-estradiol (E2) receptor isoforms [estrogen receptor (ER) alpha and ERbeta] bind E2 and selective ER modulators (SERMs) as homodimers (alpha/alpha or beta/beta) or heterodimers (alpha/beta) to regulate gene expression. Although recent studies have shown that ER homodimers regulate unique sets of E2 responsive genes, little information exists regarding the transcriptional actions of the ERalpha/beta heterodimer. This paper describes the development of a U2OS human osteosarcoma (osteoblast) cell line stably expressing both ERalpha and ERbeta isoforms at a ratio of 1:4, a ratio reported to exist in normal, mature osteoblast cells derived from cancellous bone. The regulation of endogenous genes by E2 and 4-hydroxy-tamoxifen were measured in these cells using gene microarrays and real-time RT-PCR. Both E2 and 4-hydroxy-tamoxifen were shown to regulate unique sets of endogenous genes in the U2OS-ERalpha/beta heterodimer cell line (20% and 27% of total, respectively), compared with all the genes regulated in U2OS-ER homodimer cell lines. Furthermore, two novel E2-regulated genes, retinoblastoma binding protein 1 and 7-dehydrocholesterol reductase, were found to contain estrogen response element-like sequences that directly bind the ERalpha/beta heterodimer. These results suggest that the expression of both ER isoforms, forming functional ERalpha/beta heterodimers, result in unique patterns of gene regulation, many of which are distinct from the genes regulated by the ER homodimers. PMID- 15802377 TI - Proper interpretation of non-differential misclassification effects: expectations vs observations. AB - BACKGROUND: Many investigators write as if non-differential exposure misclassification inevitably leads to a reduction in the strength of an estimated exposure-disease association. Unfortunately, non-differentiality alone is insufficient to guarantee bias towards the null. Furthermore, because bias refers to the average estimate across study repetitions rather than the result of a single study, bias towards the null is insufficient to guarantee that an observed estimate will be an underestimate. Thus, as noted before, exposure misclassification can spuriously increase the observed strength of an association even when the misclassification process is non-differential and the bias it produced is towards the null. METHODS: We present additional results on this topic, including a simulation study of how often an observed relative risk is an overestimate of the true relative risk when the bias is towards the null. RESULTS: The frequency of overestimation depends on many factors: the value of the true relative risk, exposure prevalence, baseline (unexposed) risk, misclassification rates, and other factors that influence bias and random error. CONCLUSIONS: Non-differentiality of exposure misclassification does not justify claims that the observed estimate must be an underestimate; further conditions must hold to get bias towards the null, and even when they do hold the observed estimate may by chance be an overestimate. PMID- 15802378 TI - Seroprevalence of hepatitis C virus and associated risk behaviours: a population based study in San Juan, Puerto Rico. AB - BACKGROUND: Limited information about the epidemiology of hepatitis C virus (HCV) infection is available in Puerto Rico, one of the areas hardest hit by the human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) epidemic. We estimated the prevalence of HCV infection and identified correlates of seropositivity in the municipality of San Juan, Puerto Rico. METHODS: A probability cluster design was employed to select a sample of households representative of the population aged 21-64 years in San Juan during 2001-2002. All 964 subjects completed a face-to-face interview to gather data on demographics and self-reported risk behaviours followed by venipuncture for HCV antibody testing. Variables that were at least marginally associated with HCV seroprevalence (P < 0.10) in the bivariate analyses were considered for inclusion into the multiple logistic regression model to estimate the adjusted prevalence odds ratio (POR). RESULTS: Overall weighted prevalence of HCV infection was 6.3% (95% CI 3.6-10.9%). A significant (P < 0.05) higher prevalence was observed among subjects with the following characteristics: age 30-49 (9.5%), male sex (10.6%), < or =12 years of education (9.6%), no health coverage (12.6%), lifetime heroin use (39.2%), lifetime cocaine use (39.6%), tattooing practices (34.2%), history of imprisonment (32.8%), and self-reported histories of hepatitis B virus infection (30.4%) and HIV/AIDS (92.1%). Multivariate logistic regression revealed that tattooing practices (POR = 8.9; 95% CI 1.7-44.7), lifetime cocaine use (POR = 5.5; 95% CI 2.2-13.5), blood transfusions prior to 1992 (POR = 4.0; 95% CI 1.6 10.1), lifetime heroin use (POR = 3.3; 95% CI 1.4-7.8), and history of imprisonment (POR = 2.3; 95% CI 1.1-4.9) remained significantly associated with HCV seropositivity. CONCLUSIONS: The large prevalence of HCV infection observed in Puerto Rican adults residing in San Juan suggest that HCV infection is an emerging public health concern and merits further investigation. PMID- 15802379 TI - Long-term risk of tuberculosis among immigrants in Norway. AB - BACKGROUND: Two-thirds of the tuberculosis (TB) cases in Norway were discovered among immigrants. Some cases were discovered at arrival, but many develop the disease several years post-migration. Knowledge about how long after migration to Norway TB were discovered will enable us to better target preventive measures including preventive therapy. This study examines the long-term risk of TB among immigrants in Norway. METHODS: All non-Nordic immigrants who arrived in Norway between 1986 and 2002, as registered by the Norwegian Directorate of Immigration, were followed-up. Their TB status was determined from the National Tuberculosis Registry. Observation period for TB cases was calculated from the date of arrival in Norway to TB registration. For persons without TB, it was calculated from the date of arrival in Norway to the date of emigration from Norway, date of death, or until end of follow-up (December 31, 2002). RESULTS: For immigrants from Africa and Asia, the TB rates were 190 and 80 per 100,000 person-years (PY), respectively, at 7 years post-migration. For immigrants from Somalia, Pakistan, Vietnam, and the former Yugoslavia, the rates were 520, 160, 210, and 40 per 100 000 PY respectively, at 7 years post-migration. These rates were 7 to 90 times higher than the crude TB incidence for Norway. This increased risk applies to both genders, pulmonary and extra-pulmonary sites. CONCLUSION: These results indicate the need for health personnel to be aware that immigrants remain at high risk of TB many years post-migration. Screening for TB on arrival should be strengthened, and preventive therapy for those with recent TB infection should be considered. PMID- 15802380 TI - When aspirations and achievements don't meet. A longitudinal examination of the differential effect of education and occupational attainment on declines in self rated health among Canadian labour force participants. AB - BACKGROUND: To examine the association of a mismatch between educational qualifications and occupational attainment and subsequent declines in self-rated health (SRH) in a longitudinal nationally representative Canadian population sample. METHODS: This study used longitudinal data from 4045 healthy, working respondents of the Canadian National Population Health Survey. Respondents were categorized as either qualified, overqualified, or underqualified based on the match between their education and the skills required for their current occupation over a 2-year period. Logistic regression analysis estimated the odds of decline in SRH over the following 4-year period, using the match between occupation and education as the main independent variable. Analyses were controlled for a number of confounding variables including health behaviours, mental health, self-esteem, job control, and demographic information. RESULTS: Relative to respondents with university education working in occupations for which they were qualified, respondents with university education, working in occupations for which they were overqualified had a significant risk of decline in SRH between 1996 and 2000, even after adjusting for a number of potential confounders (OR = 2.08, 95% CI 1.11-3.91). In respondents with secondary education or less, differences in occupational attainment were not associated with differences in the odds of decline in SRH. CONCLUSIONS: The effect of occupational attainment on health is important for individuals who have invested the most time in their education. Conversely, differential occupational attainment is not associated with differences in the odds of decline in health for participants with lower levels of education. PMID- 15802381 TI - Commentary: estimating and understanding area health effects. PMID- 15802382 TI - Commentary: modern day 'flying ambulances' for coronary care: a tale of two cities. PMID- 15802383 TI - Pre-hospital coronary care and coronary fatality in the Belfast and Glasgow MONICA populations. AB - BACKGROUND: The aim of this study was to describe and compare coronary event case fatality and care pathways in two defined populations with access to different models of pre-hospital care provision. METHODS: Secondary analysis of MONItoring of Trends and Determinants in CArdiovascular Disease (MONICA) population coronary event registers (1988, 1989, 1990, 1992 and 1993). RESULTS: Case fatality at 28 days following an acute coronary event was 6.5% greater in the Glasgow MONICA Project (GMP) population (46.7%) than in the Belfast MONICA Project (BMP) population (40.2%). Pre-hospital case fatality was 33.9% in the GMP population and 28.3% in the BMP population. These differences could not be fully explained by mobile coronary care unit (MCCU) responses in the BMP area. Initial care was provided in hospital for 28.3% of the BMP events and only 7.7% of the GMP events. Additional data collected by the Belfast and Glasgow MONICA investigators support a large difference between the median delay to main medical care in the BMP events (120 min) and the median delay to ward admission in the GMP area (220 min) at this time. CONCLUSIONS: Our findings suggest that the delay between coronary event onset and access to specialist coronary care was the most likely critical difference, irrespective of hospital-based MCCU provision in the BMP area. An established 'culture of early intervention' in Belfast may have been an important factor. As a large proportion of coronary event fatalities continue to occur outside hospital, there is a need to strengthen the evidence base underpinning the provision of appropriate skilled care and treatment at the earliest possible opportunity. PMID- 15802384 TI - Identification of HMG-CoA reductase inhibitors as activators for human, mouse and rat constitutive androstane receptor. AB - Constitutive active (or androstane) receptor (CAR, NR1I3), a member of the nuclear receptor family, is a major regulator for induction of cytochrome P450 2B (CYP2B) genes by phenobarbital. Phenobarbital-like inducer, 1,4-bis[2-(3,5 dichloropyridyloxy)]benzene is a potent mouse CAR ligand that has been used to study CAR target genes in mice but does not activate human CAR (hCAR) or rat CAR (rCAR). Although 6-(4-chlorophenyl) imidazo[2,1-b][1,3]thiazole-5-carbaldehyde O (3,4-dichlorobenzyl)oxime (CITCO) was reported to be an hCAR agonistic ligand, activation of hCAR by CITCO in cell-based reporter assay was weak. Therefore, we performed a screening of 50 drugs and chemicals using cell-based reporter assays to identify activators of hCAR. Among them, HMG-CoA reductase inhibitors (cerivastatin, simvastatin, fluvastatin, and atorvastatin) enhanced the hCAR mediated transcriptional activation of phenobarbital-responsive enhancer module reporter gene by up to 3-fold. Similar activation by HMG-CoA reductase inhibitors was also observed with mouse and rat CARs. On the other hand, pravastatin did not activate hCAR at the concentrations tested (up to 30 microM). The extent of activation by the HMG-CoA reductase inhibitors was stronger than that by CITCO. Cerivastatin, simvastatin, fluvastatin, and atorvastatin induced CYP2B6 mRNA in stable hCAR-expressed FLC7 cells but not in original FLC7 cells. Therefore, we concluded that CAR mediates the effects of HMG-CoA reductase inhibitors on the induction of CYP2B genes, although HMG-CoA reductase inhibitors also activate pregnane X receptor. HMG-CoA reductase inhibitors such as cerivastatin would be useful to study for elucidating molecular and cellular mechanisms of hCAR. PMID- 15802385 TI - Metabolic profile of 1,5-dicaffeoylquinic acid in rats, an in vivo and in vitro study. AB - To explore the metabolism of 1,5-dicaffeoylquinic acid (1,5-DCQA) in rats, liquid chromatography-mass spectrometry in parallel to diode-array detection was used for the rapid detection/characterization of the metabolites formed in bile, urine, and plasma of rats following oral administration of 1,5-DCQA (160 mg/kg). The methylation and glucuronidation of 1,5-DCQA occurring in vitro using rat liver and small intestinal microsomes and cytosols were studied in comparison with those occurring in vivo, and the enzymes involved were also determined. In addition, the anti-HIV (human immunodeficiency virus) activity of three important metabolites was preliminarily evaluated in MT-4 cells infected with HIV-1. A total of 22 metabolites in vivo and in vitro were identified, including four isomeric O-mono-methylated metabolites (M8-M11), nine isomeric O-di-methylated metabolites (M3, M6, M22, and M12-M17), four isomeric O-mono-methyl glucuronidated metabolites (M2 and M19-M21), four isomeric O-di-methyl glucuronidated metabolites (M1, M4, M5, and M7), and one glucuronidated metabolite (M18). The O-methylation positions of three important metabolites (M8, M9, and M12) were determined (3''-, 3'-, and 3',3''-) by comparing with synthesized standards. The efficacy experiments showed that M8, M9, and M12 could inhibit HIV replication with IC(50) values of about 25, 25, and 46 microM, respectively. These results suggest that O-methylation and glucuronidation are two important metabolic pathways of 1,5-DCQA, that both rat liver and small intestine can catalyze such reactions by catechol-O-methyltransferase and UDP glucuronosyltransferases, and that the HIV-1 inhibitory activity of M8, M9, and M12 is comparable to or slightly weaker than that of 1,5-DCQA. PMID- 15802386 TI - The use of a substrate cassette strategy to improve the capacity and throughput of cytochrome P450 induction studies in human hepatocytes. AB - The gold standard for estimating the ability of a new chemical entity to induce the human cytochromes P450 (P450s) is to determine the increase in catalytic activity of these enzymes after treatment of primary cultures of human hepatocytes with the potential inducer. The limited availability of fresh human hepatocytes makes these studies impossible to perform on demand. A substrate cassette strategy in which probe substrates for three different P450s were simultaneously added to hepatocyte preparations was studied to determine whether this would be a viable method to increase the capacity of induction studies. The biotransformations of phenacetin (CYP1A2), diclofenac (CYP2C9), and midazolam (CYP3A) were compared when administered to four different hepatocyte preparations (+/-inducer) individually or simultaneously as a cassette. The determinations of fold-induction in response to known inducers were not significantly affected, although slight differences were occasionally observed between the various P450 activities, whether determined individually or in the cassette. In total, for three P450 activities in the four hepatocyte preparations with and without inducer, no trend demonstrating a drug-drug interaction among any of the three probes was detected. The lack of interactions between the probe substrates demonstrates that this cassette strategy may be used in primary human hepatocyte induction studies without concern that the interactions between the substrates may be affecting the results, as has been seen in other cassette dosing experiments. Therefore, this substrate cassette is an excellent method for increasing the capacity and throughput of human hepatocyte induction studies by combining three experiments into one. PMID- 15802387 TI - Kinetic characterization of the 1A subfamily of recombinant human UDP glucuronosyltransferases. AB - The initial glucuronidation rates were determined for eight recombinant human UDP glucuronosyltransferases (UGTs) of the 1A subfamily, and the bisubstrate kinetics and inhibition patterns were analyzed. At low substrate concentrations, the reactions followed general ternary complex kinetics, whereas at higher concentrations of both substrates, the reactions were mostly characterized by ternary complex kinetics with substrate inhibition. The glucuronidation of entacapone by UGT1A9 was inhibited by 1-naphthol in a competitive fashion, with respect to entacapone, and an uncompetitive fashion, with respect to UDP glucuronic acid (UDPGA). Its inhibition by UDP, on the other hand, was noncompetitive with respect to entacapone and competitive with respect to UDPGA. These inhibition patterns are compatible with a compulsory ordered bi bi mechanism in which UDPGA is the first-binding substrate. Despite the identical primary structure of the C-terminal halves of the UGT1A isoforms, there were marked differences in the respective K(m) values for UDPGA, ranging from 52 microM for UGT1A6 to 1256 microM for UGT1A8. Relative specificity constants were calculated for the eight UGT1A isoforms with 1-hydroxypyrene, 4-nitrophenol, scopoletin, 4-methylumbelliferone, and entacapone as aglycone substrates. The results demonstrated that seven of the UGT1A isoforms are capable of conjugating phenolic substrates with similar highest k(cat) values, and UGT1A4 has a lower relative turnover rate. The highest specificity constants were obtained for 1 hydroxypyrene, even with UGT1A6, which has been regarded as a specific isoform for small planar phenols. A k(cat) value of 1.9 s(-1) was calculated for the glucuronidation of scopoletin by purified UGT1A9. PMID- 15802388 TI - Tissue distribution and hepatic and renal ontogeny of the multidrug resistance associated protein (Mrp) family in mice. AB - Analysis of the mouse genome has revealed eight multidrug resistance-associated (Mrp) transporters, with mouse homologs for all human MRPs except MRP8. Whereas MRP expression in tissues of humans and rats has been examined, no characterization exists for mice. Furthermore, the ontogeny of mouse Mrps is unknown, and such knowledge may be helpful in understanding age-related pharmacokinetics. Therefore, the purpose of this study was to quantitatively determine 1) expression of the Mrp family in 12 different tissues, 2) gender variations in Mrp expression in liver and kidney, and 3) whether Mrp expression is altered during development. Highest expression of the Mrp family members is as follows: Mrp1 in testes, ovary, and placenta; Mrp2 in intestine, followed by liver and kidney; Mrp3 in large intestine; Mrp4 in kidney; Mrp5 in brain, followed by lung and stomach; Mrp6 in liver; Mrp7 in testes, intestine, and kidney; and Mrp9 solely in testes. Gender differences in Mrp expression were observed: Mrp1, 3, and 4 in kidney, as well as Mrp1 and 4 in liver were female predominant. Ontogeny of the four Mrps expressed in liver was as follows: Mrp2 and Mrp4 were expressed at adult levels at birth; Mrp3 reached adult levels at day 30, and Mrp6 was not expressed until day 10. In kidney, Mrp1 and Mrp5 were expressed at adult levels at birth, whereas Mrp2, 3, 4, and 6 generally increased over time. In conclusion, marked differences in expression of the individual Mrp family members exist in various tissues, with age, and with gender. PMID- 15802389 TI - Expression and induction potential of cytochromes P450 in human cryopreserved hepatocytes. AB - Fresh human hepatocytes are still considered as the "gold standard" to screen in vitro for cytochrome P450 (P450) induction. However, sparse availability of good quality human liver tissue for research purposes and the demand for standardized cell populations, together with the need for proper storage of the cells not immediately required, have resulted in the development of cryopreservation techniques that provide adequate viability and plateability of hepatocytes after thawing. This study aimed at validating cryopreserved human hepatocytes as a model to investigate P450 induction. Cryopreserved cells from four different donors were plated and cultured for 48 h, followed by incubation in the presence of typical P450 inducers. During the experiments, quality of the cultured cells was monitored both physiologically and morphologically. Concomitantly, the activity of CYP1A2, 2B6, 2C9, 2E1, and 3A4 was measured together with their mRNA and protein expression. Determination of CYP1A2, 2B6, 2C9, 2E1, and 3A4 activity in control versus prototypical inducer-treated hepatocytes revealed a maximal significant mean 11.6-, 2.8-, 1.9-, 1.5-, and 9.0-fold induction over their basal expression, respectively. Protein expression analysis of these P450s confirmed these results. Moreover, a mean 44.9-, 3.5-, 3.2-, and 13.8-fold induction of CYP1A2, 2B6, 2C9, and 3A4 mRNA was observed. Our data demonstrate that cryopreserved human hepatocytes are a valuable tool to study the induction of CYP1A2, 2B6, 2C9, 2E1, and 3A4. PMID- 15802391 TI - Induction of labor in the nulliparous gravida with an unfavorable cervix. PMID- 15802392 TI - Bishop score and risk of cesarean delivery after induction of labor in nulliparous women. AB - OBJECTIVE: To quantify the risk and risk factors for cesarean delivery associated with medical and elective induction of labor in nulliparous women. METHODS: A prospective cohort study was performed in nulliparous women at term with vertex singleton gestations who had labor induced at 2 obstetrical centers. Medical and elective indications and Bishop scores were recorded before labor induction. Obstetric and neonatal data were analyzed and compared with the results in women with a spontaneous onset of labor. Data were analyzed using univariate and multivariable regression modeling. RESULTS: A total of 1,389 women were included in the study. The cesarean delivery rate was 12.0% in women with a spontaneous onset of labor (n = 765), 23.4% in women undergoing labor induction for medical reasons (n = 435) (unadjusted odds ratio [OR] 2.24; 95% confidence interval [CI] 1.64-3.06), and 23.8% in women whose labor was electively induced (n = 189) (unadjusted OR 2.29; 95% CI 1.53-3.41). However, after adjusting for the Bishop score at admission, no significant differences in cesarean delivery rates were found among the 3 groups. A Bishop score of 5 or less was a predominant risk factor for a cesarean delivery in all 3 groups (adjusted OR 2.32; 95% CI 1.66 3.25). Other variables with significantly increased risk for cesarean delivery included maternal age of 30 years or older, body mass index of 31 or higher, use of epidural analgesia during the first stage of labor, and birth weight of 3,500 g or higher. In both induction groups, more newborns required neonatal care, more mothers needed a blood transfusion, and the maternal hospital stay was longer. CONCLUSION: Compared with spontaneous onset of labor, medical and elective induction of labor in nulliparous women at term with a single fetus in cephalic presentation is associated with an increased risk of cesarean delivery, predominantly related to an unfavorable Bishop score at admission. LEVEL OF EVIDENCE: II-2. PMID- 15802393 TI - Labor progression and risk of cesarean delivery in electively induced nulliparas. AB - OBJECTIVE: To describe the pattern of labor progression and risk of cesarean delivery in women whose labor was electively induced. METHODS: We analyzed data on all low-risk, nulliparous women with an elective induction or spontaneous onset of labor between 37 + 0 and 40 + 6 weeks from January 2002 to March 2004 at a single institution. The median duration of labor by each centimeter of cervical dilation and the risk of cesarean delivery were computed for 143 women with preinduction cervical ripening and oxytocin induction, 286 women with oxytocin induction, and 1,771 women with a spontaneous onset of labor. An intracervical Foley catheter was used to ripen the cervix. RESULTS: Electively induced labor with cervical ripening had substantially slower latent and early active phases. After controlling for potential confounders, women who had an elective induction with cervical ripening had 3.5 times the risk of cesarean delivery during the first stage of labor (95% confidence interval 2.7-4.5), compared with those admitted in spontaneous labor. Elective induction without cervical ripening, on the other hand, was associated with a faster labor progression from 4 to 10 cm (266 compared with 358 minutes, P < .01) and did not increase the risk of cesarean delivery, compared with those in spontaneous labor. CONCLUSION: The pattern of labor progression differs substantially for women with an electively induced labor compared with those with spontaneous onset of labor. Furthermore, elective induction in nulliparous women with an unfavorable cervix has a high rate of labor arrest and a substantially increased risk of cesarean delivery. LEVEL OF EVIDENCE: II-2. PMID- 15802394 TI - When has an induction failed? AB - OBJECTIVE: To evaluate the length of the latent phase that, during labor inductions in nulliparous women, is associated with significantly decreased chance of vaginal delivery and increased risk of maternal and neonatal morbidity. METHODS: All inductions of labor during a 6-month period were identified. Only those women who were nulliparous with a pregnancy of 36 weeks or more of gestation underwent further data analysis. Demographic data, intrapartum course, and maternal and neonatal outcomes were abstracted from the medical record. The latent phase was defined as beginning after oxytocin had been initiated and amniotomy performed and continuing until either 4 cm cervical dilation and 80% effacement or 5 cm cervical dilation regardless of effacement. RESULTS: A total of 397 nulliparous women, 32% of whom underwent cervical ripening, presented during the study period. Only 8 women (2%) never achieved active phase labor before cesarean, and the overall cesarean rate was 26.0%. A longer latent phase was associated with a greater rate of cesarean delivery, although only after 18 hours did a majority of induced labors result in cesarean. Chorioamnionitis and postpartum hemorrhage were more frequent with latent phases of labor greater than 18 hours (16% and 26%, respectively), although these diagnoses did not translate into greater risk of transfusion, hysterectomy, or prolonged hospitalization. Neonatal outcomes, including meconium passage, fetal acidemia, neonatal intensive care unit admission, or other morbidity did not increase in conjunction with longer latent phases. CONCLUSION: A latent phase of as long as 18 hours during induction of labor in nulliparous women allows the majority of these women to achieve a vaginal delivery without being subject to a significantly increased risk of significant maternal or neonatal morbidity. LEVEL OF EVIDENCE: II-2. PMID- 15802395 TI - Racial differences in cesareans: an analysis of U.S. 2001 National Inpatient Sample Data. AB - OBJECTIVE: To evaluate racial differences in potentially unnecessary cesareans in the United States. METHODS: The 2001 Healthcare Cost and Utilization Project Nationwide Inpatient Sample database was used to analyze various maternal demographic and clinical parameters among women having cesarean deliveries. For the purpose of our study, if there were no diagnoses related to cesarean delivery on the discharge certificate, the cesarean was classified as potentially unnecessary RESULTS: Using our methodology, 11% of 540,174 primary and 65% of 371,863 repeat cesareans for the year 2001 in the United States were classified as potentially unnecessary. After controlling for age, payment source, ZIP income, admission day, region of the country, and hospital size, location, and teaching status, black women had a higher likelihood of having potentially unnecessary primary cesareans when compared with white (P < .001) or Hispanic women (P < .001). White women had a higher likelihood of having potentially unnecessary repeat cesareans than black women (P < .001), although the magnitude of the odds ratio for race was not as striking as in primary cesarean. Potentially unnecessary primary cesareans were also more likely to occur in women aged 35 years or older, Medicare patients, weekend admissions, or those residing in the South or Northeast. Potentially unnecessary repeat cesareans were more likely to occur in women younger than 35 years, in rural hospitals, and in weekday admissions. CONCLUSION: In the United States, racial differences exist in the proportion of cesarean deliveries that are potentially unnecessary. Further exploration of these differences is warranted. LEVEL OF EVIDENCE: III. PMID- 15802396 TI - Routine appendectomy in epithelial ovarian carcinoma: is it necessary? AB - OBJECTIVE: To detect risk factors for the appendiceal metastasis and to define the role of routine appendectomy in patients with epithelial ovarian carcinoma. METHODS: A total of 285 patients with epithelial ovarian carcinoma who had undergone primary cytoreductive surgery including appendectomy were retrospectively evaluated. Appendiceal involvement was divided into 2 groups: gross and microscopic. Clinicopathologic variables were evaluated for possible significance in terms of appendiceal metastasis. A second analysis was performed using the same variables to detect a possible relation with microscopic metastasis. In a subgroup analysis, we also analyzed the role of routine appendectomy in patients with clinically early stage disease. RESULTS: One hundred six patients were found to have appendiceal metastasis (37%). Univariate and multivariate analysis revealed stage of disease as the unique factor determining the appendiceal metastasis (P < .001). Five patients with apparently stage I-II disease were upstaged due to isolated appendiceal metastasis (4.9%). In the second analysis excluding the patients with gross involvement, ascites was an independent predictor of microscopic involvement (P < .01). CONCLUSION: Routine appendectomy is indicated in all epithelial ovarian carcinoma patients as part of the initial surgical staging procedure because of a considerable rate of upstaging in early stage disease and optimal cytoreduction in advanced stages. LEVEL OF EVIDENCE: II-3. PMID- 15802397 TI - Antidepressant medication use [corrected] and risk of ovarian cancer. AB - OBJECTIVE: It has been hypothesized that antidepressants may enhance cancer growth. Previous studies of antidepressant use and ovarian cancer have been inconsistent and have been limited in their ability to examine the association with selective serotonin reuptake inhibitors (SSRIs), which are currently the antidepressants most commonly prescribed. The objective of this paper was to evaluate whether women with ovarian cancer were more likely to report past use of antidepressants than control women. METHODS: Antidepressant use was assessed in a population-based, case-control study of ovarian cancer (593 cases, 628 controls). Unconditional logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) associated with antidepressant use overall and by subcategories of antidepressants. RESULTS: Antidepressant use was reported by 18% of cases and 20% of controls. No increased risk was observed for ever use of any type of antidepressant (OR 0.9, 95% CI 0.7-1.2) or for SSRIs (OR 1.0, 95% CI 0.7 1.5). There also was no evidence of increased risk with longer duration of use. Our study had greater than 80% power to detect an OR as small as 1.5. Thus, even a modest increase in risk associated with antidepressant use can be excluded with these data. CONCLUSION: Our study adds to the growing body of evidence suggesting that antidepressants do not have a significant effect on ovarian cancer risk. In particular, the data suggest that SSRIs, which are the most commonly used class of antidepressants, are not associated with an increased risk for ovarian cancer. LEVEL OF EVIDENCE: II-2. PMID- 15802398 TI - Androgenic progestins in oral contraceptives and the risk of epithelial ovarian cancer. AB - OBJECTIVE: Oral contraceptives (OCs) have been consistently linked to reduced risk of ovarian cancer. Oral contraceptive formulations display varying degrees of androgenicity. Data linking androgens to ovarian cancer suggest that OC androgenicity may impact efficacy in preventing ovarian cancer. The authors investigated whether OC efficacy might differ according to androgenicity by using data from a large, population-based, case-control study (the Steroid Hormones and Reproductions [SHARE] Study). METHODS: Detailed data on OC formulation was obtained by an in-person interview for 568 cases and 1,026 controls. Multivariable logistic regression was used to assess the association of OC androgenicity with ovarian cancer while controlling for the known potential confounders of age, parity, family history of ovarian cancer, and tubal ligation. RESULTS: Androgenic and nonandrogenic OCs conferred a similar and significant reduction in ovarian cancer risk (odds ratio 0.52, 95% confidence interval 0.35 0.76 and odds ratio 0.59, 95% confidence interval 0.45-0.78, respectively). No differences in duration of use, age at first use, and time since last use were found between androgenic and nonandrogenic formulations. CONCLUSION: In general, the androgenicity of an OC does not alter chemopreventive efficacy. LEVEL OF EVIDENCE: II-2. PMID- 15802399 TI - Atypical squamous cells of undetermined significance: human papillomavirus testing in adolescents. AB - OBJECTIVE: To estimate the age-stratified prevalence of oncogenic human papillomavirus (HPV) infection and to evaluate risk factors for HPV acquisition among women with atypical squamous cells of undetermined significance (ASC-US). METHODS: This was a chart review of all women with ASC-US smears who underwent oncogenic HPV testing between July 2002 and February 2004. To be eligible, HPV DNA results had to be available. Data extracted from charts included demographic information as well as certain patient characteristics historically associated with HPV acquisition or carriage. RESULTS: Of 527 eligible women with ASC-US, 357 (68%, 95% confidence interval [CI] 64-72%) tested positive for oncogenic HPV. As compared with women who tested negative, this population was significantly younger and less likely to be married. When stratified by age, 77% of the women aged younger than 20 years were positive for high-risk subtypes, compared with 58% of women aged older than 25 years (P < .01). In the multivariate analysis, young age (relative risk [RR] 1.30, 95% CI 1.14.-1.49), current smoking (RR 1.14, 95% CI 1.03-1.25) and a history of chlamydial infection (RR 1.20, 95% CI 1.09 1.32) were associated with an increased likelihood of HPV infection, while oral contraceptive use (RR 0.86, 95% CI 0.72-1.03) had a marginal protective effect. The protective effect of oral contraceptives was most pronounced among adolescent women (RR 0.57, 95% CI 0.31-1.03). CONCLUSION: Given that the rate of oncogenic HPV infection approached 80% in our population of adolescent women with ASC-US, the usefulness of HPV testing in this age group requires further investigation. LEVEL OF EVIDENCE: II-3. PMID- 15802400 TI - Patient gender preferences in a large military teaching hospital. AB - OBJECTIVE: To quantify preferences for female providers among obstetric gynecology patients in a large military teaching hospital in California. METHODS: A questionnaire regarding patient preferences for female provider was distributed to and collected from 1,544 obstetric-gynecology patients over a two-month period. Responses were tabulated overall and for several demographic subgroups. RESULTS: Sixty percent of respondents indicated they had no provider gender preference or they preferred a male. Significant ethnic preferences were noted among Asian, Pacific Islander, and Native-American women. Junior officers, wives, and daughters of service members also showed a greater preference for a female physician provider. CONCLUSION: The majority of obstetric-gynecology patients surveyed had no strong preferences for female providers. In the interest of patient satisfaction, where strong gender preferences exist, attempts to accommodate could be made without overburdening female staff. LEVEL OF EVIDENCE: III. PMID- 15802401 TI - A national estimate of the elective primary cesarean delivery rate. AB - OBJECTIVE: We describe national trends for elective primary cesarean delivery from 1994 to 2001, with attention to changes in indications. METHODS: We used data from the Healthcare Cost and Utilization Project Nationwide Inpatient Sample. Cesarean deliveries were identified by International Classification of Diseases, 9th Revision, Clinical Modification procedure and diagnostic codes; V codes identified all types of deliveries for denominators. Twelve indications for elective primary cesarean delivery were targeted. International Classification of Diseases, 9th Revision, Clinical Modification coding changes were also evaluated. RESULTS: After excluding women who had labored and previous cesarean deliveries, elective primary cesarean deliveries rose from 19.7% of all cesarean deliveries in 1994 to 28.3% in 2001, an increase of approximately 43.6%. The use of the identified indications for elective primary cesarean delivery increased for codes representing malpresentation, antepartum bleeding, hypertension and severe hypertension, macrosomia, unengaged head, preterm gestation, and maternal soft tissue disorders. Coding for herpes, multiple gestation, other uterine scar, and congenital central nervous system remained the same. Additionally, a new 1998 code for fetal heart rate abnormalities was rapidly adopted during the study period. CONCLUSION: A national estimate of the elective primary cesarean delivery rate shows a rising trend. Additionally, coded indications for these procedures are shifting. Further examination into the use and clinical implications of indications through national surveillance for elective primary cesarean delivery is important for future obstetric practice. A revision of the terminology classification used to identify indications for cesarean delivery procedures would aid in this effort. LEVEL OF EVIDENCE: III. PMID- 15802402 TI - Effects of folic acid fortification on twin gestation rates. AB - OBJECTIVE: Previous studies have reported an increase in twinning of as much as 40% associated with folic acid-containing supplements, and folic acid fortification of enriched cereal grains was authorized in 1996. The purpose of this study was to investigate whether twinning rates have increased since that time. METHODS: We used United States birth and fetal death records to calculate twin gestation rates from 1990 through 2000. To eliminate the influence of fertility treatments, our analysis was limited to nulliparous women aged 16-19. We compared time trends in twin gestation rates before and after folic acid fortification in 1996. RESULTS: A total of 25,065 twin and 3,362,245 singleton pregnancies were included. Twin gestation rates were stable from January 1990 through December 1996, at 7.2 per 1,000, and then began a steady increase, averaging 2.4% (95% confidence interval 0.1-4.2%, P = .006) per year, which continued through 2000 and reached 8.2 per 1,000. This translates to 2 additional twin pregnancies per 10,000 gestations per year. Twin rates continued to increase well beyond 1998, when the maximal fortification effect on folate status had been reached. CONCLUSION: Although twin gestation rates in women not using fertility treatments increased after food fortification with folic acid, they rose by much less than the 40% rate previously reported; the observed pattern of increase in twin gestation rates is not consistent with a folic acid fortification effect. LEVEL OF EVIDENCE: II-2. PMID- 15802403 TI - Effect of fetal position on second-stage duration and labor outcome. AB - OBJECTIVE: To evaluate the effect of fetal position on 1) second-stage labor duration and 2) indicators of maternal and neonatal morbidity. METHODS: A retrospective cohort study was conducted using a database from a previously reported randomized clinical trial. The data set includes 210 women with the fetus in a posterior position, 200 women with the fetus in a transverse position, and 1,198 women with the fetus in an anterior position. Mean durations of the second stage of labor for different fetal positions were compared using Tukey studentized test. A multivariate logistic regression model was performed to examine the determinants of prolonged second-stage duration (>or= 3 hours). Kaplan-Meier survival curves were used to graph and compare the duration of the second stage of labor for spontaneous delivery according to the fetal position at full dilatation and study group. RESULTS: Fetal malposition at full dilatation was associated with a significantly increased risk of instrumental vaginal delivery, cesarean delivery, oxytocin administration before full cervical dilatation, episiotomy, severe perineal laceration, and maternal blood loss of more than 500 mL (all P values < .01). Compared with the occiput anterior positions, there were significant differences in the duration of the second stage of labor, with a mean of 3.1 hours (95% confidence interval [CI] 3.0-3.2) for occiput anterior positions, 3.6 hours (95% CI 3.3-3.9) for occiput transverse positions (P < .05), and 3.8 hours (95% CI 3.5-4.1) for occiput posterior positions (P < .05) in the delayed pushing group. For the early pushing group, means were 2.2 hours (95% CI 2.1-2.3) for occiput anterior positions, 2.5 hours (95% CI 2.3-2.8) for occiput transverse positions (P < .05), and 3.0 hours (95% CI 2.7-3.3) for occiput posterior positions (P < .05). CONCLUSION: Fetal malposition at full dilatation results in a higher risk of prolonged second stage of labor and increases maternal morbidity indicators. LEVEL OF EVIDENCE: II-2. PMID- 15802404 TI - Improving the accuracy of fetal foot length to confirm gestational duration. AB - OBJECTIVE: To establish normative fetal foot length ranges using last menstrual period (LMP) and ultrasound dating by biparietal diameter and to examine variations in these ranges by ethnicity. METHODS: A consecutive series of 1,099 eligible subjects receiving abortions had fetal foot lengths measured directly. Models of fetal foot length were developed by using assessment of gestational duration by LMP alone, ultrasonography alone, and "best estimate" (LMP confirmed by ultrasonography). RESULTS: The full sample model using ultrasound dating (n = 1,099) yielded the following equation: foot length = -30.3 + days of gestation x 0.458 (R(2) of 0.92). Regression by LMP-determined gestational duration by using the "best estimate" sample (n = 491) provided an almost identical equation (foot length = -29.8 + days of gestation x 0.45) and a similar R(2) value of 0.87, although the standard errors were larger. Gestational duration by ultrasonography alone produced a better model fit than duration by LMP alone. Regressions by ethnicity were not significantly different compared with the simple regression, regardless of method used to determine gestational duration. CONCLUSION: A reconsideration of fetal foot length measurements to confirm gestational duration is important. More accurate tables of these measurements allow for greater precision in correlating gestational duration and foot length. Fetal foot length tables using ultrasonographically confirmed gestational duration and current statistical standards should replace tables currently used. Biparietal diameter as a single measurement provides adequate estimation of gestational duration in the second trimester for pregnancy termination, proving more reliable than LMP dating. LEVEL OF EVIDENCE: II-2. PMID- 15802405 TI - Uterine and vaginal effects of unopposed ultralow-dose transdermal estradiol. AB - OBJECTIVE: To investigate uterine effects of unopposed ultralow-dose transdermal estradiol administered to postmenopausal women for 2 years. METHODS: Postmenopausal women (n = 417), aged 60-80 years, with a uterus and with bone mineral density that was normal for age (z score >or=-2.0) were randomly assigned to receive unopposed transdermal estradiol (14 microg per day) or identical placebo patch. We evaluated effects on endometrial histology, vaginal bleeding, and vaginal epithelial cell maturation. RESULTS: At baseline, estradiol and placebo groups were similar in age (67 +/- 5 years) and in median baseline serum estradiol level (4.8 pg/mL, interquartile range 2.7, 8.0 pg/mL). In the estradiol group, median estradiol level increased to 8.6 pg/mL, (interquartile range 4.4, 13.9 pg/mL, P < .001). In the estradiol group, focal atypical endometrial hyperplasia developed in 1 woman, and adenosarcoma of the uterus developed in 1 woman. The placebo group had no endometrial hyperplasia. Endometrial proliferation occurred in 8.5% of the estradiol group and in 1.1% of the placebo group (P = .06). Incidence of vaginal bleeding was 12.4% in the estradiol group and 8.6% in the placebo group (P = .3). Vaginal epithelial cells showed greater maturation in the estradiol group than in the placebo group (P < .001) but less than typically observed with standard doses of estrogen. CONCLUSION: During 2 years of treatment with ultralow-dose unopposed estradiol, treatment and placebo groups had similar rates of endometrial hyperplasia, endometrial proliferation, and vaginal bleeding. This therapy apparently causes little or no endometrial stimulation. LEVEL OF EVIDENCE: I. PMID- 15802406 TI - Reduced fertilization rates in older men when cervical mucus is suboptimal. AB - OBJECTIVE: Cervical mucus is vital in the regulation of sperm survival and transport through the reproductive tract. The goal of this study is to assess whether the lowered fertility for men in their late 30s and early 40s is related to the nature of cervical mucus on the day of intercourse. METHODS: In a prospective study of 7 European family planning centers, 782 couples not using birth control recorded daily observations of intercourse and the nature of cervical mucus. Using data from 1,459 menstrual cycles, 342 ending in pregnancy, we estimate day-specific conception probabilities in relation to mucus and male and female age. RESULTS: On days where cervical mucus was not evident, intercourse for men in their late 30s and early 40s was 50% less likely to result in a clinical pregnancy, adjusting for intercourse timing and female age. As secretions become more conducive to sperm transport, the effect of male age diminishes steadily from 21% on days with damp secretions, to 11% on days with thick mucus, to only 4% on days with most fertile-type mucus. CONCLUSION: The effect of male age on fecundability can be minimized by timing intercourse on days with optimal secretions. LEVEL OF EVIDENCE: II-2. PMID- 15802407 TI - Failure to obtain desired postpartum sterilization: risk and predictors. AB - OBJECTIVE: The aim of this study was to assess the rate of and risk factors for not obtaining postpartum sterilization among women who expressed a desire for sterilization during antepartum care. METHODS: In this retrospective study, we identified a cohort of women who expressed desire for postpartum sterilization at our center between March 2002 and November 2003. We compared women who did and those who did not undergo the procedure, based on demographic, antenatal, and intrapartum factors. RESULTS: Of the 712 women who expressed desire for postpartum sterilization during antepartum care, 327 (46%) did not undergo the procedure. In multivariable analysis, women who were between the ages of 21 and 25 years (adjusted odds ratio [OR] 0.56, 95% confidence interval [CI] 0.35-0.89), were African American (OR 0.68, 95% CI 0.47-1.00), requested sterilization in the second trimester (OR 0.50, 95% CI 0.29-0.86)), and had a vaginal delivery (OR 0.21, 95% CI 0.14-0.32) rather than cesarean delivery were least likely to undergo postpartum sterilization. CONCLUSION: Despite their initial request, only 54% of women in our sample underwent sterilization. Young age, African-American race, request in the second trimester, and vaginal delivery were significantly associated with not undergoing sterilization. Our data suggest that providers should counsel all women who desire postpartum sterilization about the wide array of contraceptive methods available, with the understanding that approximately half of all women may not undergo the sterilization procedure. LEVEL OF EVIDENCE: II-2. PMID- 15802408 TI - Pelvic floor surgery in the older woman: enhanced compared with usual preoperative assessment. AB - OBJECTIVE: To examine whether knowledge of deficits obtained in a preoperative geriatric assessment may benefit postoperative health outcomes in older women undergoing pelvic surgery. METHODS: This study employed a pre-post intervention cohort design. Primary outcome was difference in scores of the Physical Component Summary and Mental Component Summary of the Medical Outcomes Study Short Form 36 Health Survey in 62 older women who had undergone "usual" compared with an "enhanced" preoperative assessment consisting of Activities of Daily Living, Instrumental Activities of Daily Living, Get Up and Go Test, Draw a Clock Test, Mini Nutritional Assessment, Geriatric Depression Scale, and Social Support Scale. The assessment results were placed on the participant's hospital chart. Repeated measures analysis was used. RESULTS: There were no significant differences in Mental Component Summary scores between the usual and enhanced assessment cohorts preoperatively (mean +/- standard deviation; 49.14 +/- 10.61 compared with 53.2 +/- 9.33), at 6 weeks (53.69 +/- 8.61 compared with 55.47 +/- 9.46), or at 6 months postoperatively (53.85 +/- 10.77 compared with 56.25 +/- 7.25); P = .120 for group effect and P = .798 for group by time interaction. Significant time effect was noted (P = .036). There was no significant difference in Physical Component Summary scores between the usual and enhanced assessment cohorts with respect to group effect (P = .986); there was a significant time effect (P = < .001) and a significant group by time interaction (P = .026). Satisfaction with treatment was high in both cohorts at 6 weeks and 6 months. CONCLUSION: A preoperative geriatric assessment did not seem to have differential benefit in healthy older women undergoing elective pelvic floor surgery. LEVEL OF EVIDENCE: II-2. PMID- 15802409 TI - Fallopian tube prolapse after laparoscopic resection of pelvic endometriosis. AB - BACKGROUND: Fallopian tube prolapse is an unusual but often reported complication after hysterectomy. This problem has not yet been reported in a patient undergoing laparoscopy but not hysterectomy. CASE: Fallopian tube prolapse was diagnosed in a patient after laparoscopic excision of pelvic endometriosis, without hysterectomy. The prolapsed fallopian tube was preserved by laparoscopic retrieval from the vagina and closure of the vaginoperitoneal fistula. CONCLUSION: Laparoscopic surgery, when associated with the creation of a vaginoperitoneal fistula, is a risk factor for fallopian tube prolapse. This problem can be diagnosed and safely managed with a laparoscopic approach. PMID- 15802410 TI - Effect of a levonorgestrel intrauterine system on women with type 1 diabetes: a randomized trial. AB - OBJECTIVE: Women with diabetes need safe, effective contraception. Although intrauterine devices provide superior contraception, concerns remain that progestin absorbed systemically from the levonorgestrel-releasing device may impair carbohydrate metabolism. To examine the effect of the levonorgestrel releasing intrauterine system on glucose metabolism in diabetic women. METHODS: We randomly assigned 62 women with uncomplicated insulin-dependent diabetes mellitus to either a levonorgestrel-releasing or a copper T 380A intrauterine device. The primary outcome to assess glucose metabolism was glycosylated hemoglobin; fasting serum-glucose levels and daily insulin dose requirements over 12 months of observation were examined as well. RESULTS: Outcome data were available for 29 women using the levonorgestrel-releasing and 30 using the copper device. At 12 months, mean glycosylated levels were similar for women of the 2 groups (6.3%, standard deviation [SD] +/- 1.5 compared with 6.3%, SD +/- 1.3, respectively). The same was true for mean fasting-serum glucose levels (7.4 mM, SD +/- 4.2 compared with 7.5 mM, SD +/- 4.2) and daily insulin doses (35.1 units, SD +/- 12.8 compared with 36.4 units, SD +/- 9.0). No important differences were noted at either 6 weeks or 6 months. CONCLUSION: The levonorgestrel-releasing device had no adverse effect on glucose metabolism, even at the 6-week observation when systemic levels of levonorgestrel would have been higher than at later observations. Concern about a potential adverse effect of this contraceptive on glucose control is unwarranted, and its use in women with diabetes should be liberalized. LEVEL OF EVIDENCE: I. PMID- 15802411 TI - Differences between hospitals in cesarean rates for term primigravidas with cephalic presentation. AB - OBJECTIVE: The purpose of this study was to quantify patient populations and practice patterns at perinatal centers with the highest and lowest cesarean delivery rates. METHODS: The 2 perinatal centers in our state with the lowest (Hospital A-16.6%) and highest (Hospital B-20.3%) overall cesarean rates for Robson group 1 (term primigravidas, vertex, spontaneous labor) and group 2 (term primigravidas, vertex, induced labor) were identified. A total of 174 medical records at Hospital A and 150 records at Hospital B were reviewed. Statistical analysis was performed using independent-sample t tests, chi(2), and multiple logistic regression. RESULTS: Indications for cesarean delivery were not different between the 2 groups, with the majority being for failure to progress in labor and nonreassuring fetal status. There were no differences between groups in rates of postpartum hemorrhage, chorioamnionitis, or endometritis. There were no differences in neonatal outcomes. Although women delivering in hospital A were not more likely to receive oxytocin augmentation (P = .291), their mean maximal oxytocin dosage was higher (14.5 units compared with 11.6 units, P < .001), and they were more likely to receive both fetal scalp electrodes (60.9% compared with 37.3%, P < .001) and intrauterine pressure catheters (63.8% compared with 26.0%, P < .001). CONCLUSION: Because safe reduction in cesarean delivery rates for primigravidas will proportionately reduce the number of repeat cesarean delivery required, benchmarking practices as described in this study can be considered in obstetric practices interested in long-term reductions of their cesarean delivery rates. LEVEL OF EVIDENCE: III. PMID- 15802412 TI - Uterine electromyography characteristics for early diagnosis of mifepristone induced preterm labor. AB - OBJECTIVE: Differentiating uterine contractions leading to preterm birth from ineffective uterine activity is difficult with current tools. Uterine electromyographic activity is recordable and consists of bursts (group of action potentials) characterized by characteristics that are different during pregnancy and labor. Our aim was to identify the chronology of the changes in uterine pressure and electromyographic characteristics during mifepristone-induced preterm labor in pregnant rats and to determine the earliest characteristic to change. METHODS: On day 17 of gestation, intrauterine catheter and electromyography electrodes were implanted in the uterus. On day 18, rats were allocated for treatment with mifepristone or placebo. Intrauterine pressure and electromyography integral activities and electromyography mean were calculated before treatment and 6, 12, 18, 20, 22, and 24 hours after treatment. After mathematical transformation, burst analysis was performed by using power density spectrum energy, peak amplitude, and frequency. RESULTS: As expected, delivery rate within 24 hours was higher in the mifepristone-treated group. Changes in electromyography integral activity and mean, power density spectrum energy, and intrauterine pressure integral activity occurred late during preterm labor, in a range of 2-4 hours before delivery. Electromyography peak frequency of the power density spectrum exhibited early changes, with a shift from low to high frequencies starting at 12 hours before delivery. CONCLUSION: Electromyography peak frequency of the power density spectrum from individual bursts was the first characteristic to change after antiprogestin treatment, preceding any change in intrauterine pressure, making it a potentially useful marker for the early diagnosis of preterm labor. PMID- 15802413 TI - Risk of recurrence of anal sphincter lacerations. AB - OBJECTIVE: To estimate the rate of recurrence of anal sphincter lacerations in subsequent pregnancies and analyze the risk factors associated with recurrent lacerations METHODS: Data were obtained from the Pennsylvania Health Care Cost Containment Council, Division of In-Patient Statistics, regarding all cases of third- and fourth-degree perineal lacerations that occurred during a 2-year period (from January 1990 through December 1991). All subsequent pregnancies in this group of women over the next 10 years were identified, and the rate of recurrence of sphincter tears and risk factors for recurrence were analyzed. RESULTS: The rate of anal sphincter lacerations was 7.31% (n = 18,888) during the first 2 years of study (1990-1991). In the next 10 years, these patients with prior lacerations were delivered of 16,152 pregnancies. Of these, 1,162 were by cesarean. Among the 14,990 subsequent vaginal deliveries, 864 (5.76%) had a recurrence of a third- or fourth-degree laceration. Women with prior fourth degree lacerations had a much higher rate of recurrence than those with prior third-degree laceration (7.73% versus 4.69%). The rate for recurrent lacerations was significantly lower than the rate for initial lacerations (odds ratio 1.29, 95% confidence interval [CI] 1.2-1.4). Forceps delivery with episiotomy had the highest risk for recurrent laceration (17.7%, odds ratio 3.6, 95% CI 2.6-5.1), whereas vacuum use without episiotomy had the lowest risk (5.88%, odds ratio 1.0, 95% CI 0.6-1.7). CONCLUSION: Prior anal sphincter laceration does not appear to be a significant risk factor for recurrence of laceration. Operative vaginal delivery, particularly with episiotomy, increases the risk of recurrent laceration as it does for initial laceration. LEVEL OF EVIDENCE: III. PMID- 15802414 TI - Risk of maternal postpartum readmission associated with mode of delivery. AB - OBJECTIVE: To determine whether cesarean and operative vaginal deliveries are associated with an increased risk of maternal rehospitalization compared with spontaneous vaginal delivery. METHODS: A population-based cohort study was conducted by using the Canadian Institute for Health Information's Discharge Abstract Database between 1997/1998 and 2000/2001, which included 900,108 women aged 15-44 years with singleton live births (after excluding several selected obstetric conditions). RESULTS: A total of 16,404 women (1.8%) were rehospitalized within 60 days after initial discharge. Compared with spontaneous vaginal delivery (rate 1.5%), cesarean delivery was associated with a significantly increased risk of postpartum readmission (rate 2.7%, odds ratio [OR] 1.9, 95% confidence interval [CI] 1.8-1.9); ie, there was 1 excess postpartum readmission per 75 cesarean deliveries. Diagnoses associated with significantly increased risks of readmission after cesarean delivery (compared with spontaneous vaginal delivery) included pelvic injury/wounds (rate 0.86% versus 0.06%, OR 13.4, 95% CI 12.0-15.0), obstetric complications (rate 0.23% versus 0.08%, OR 3.0, 95% CI 2.6-3.5), venous disorders and thromboembolism (rate 0.07% versus 0.03%, OR 2.7, 95% CI 2.1-3.4), and major puerperal infection (rate 0.45% versus 0.27%, OR 1.8, 95% CI 1.6-1.9). Women delivered by forceps or vacuum were also at an increased risk of readmission (rates 2.2% and 1.8% versus 1.5%; OR forceps: 1.4, 95% CI 1.3-1.5; OR vacuum: 1.2, 95% CI 1.2-1.3, respectively). Higher readmission rates after operative vaginal delivery were due to pelvic injury/wounds, genitourinary conditions, obstetric complications, postpartum hemorrhage, and major puerperal infection. CONCLUSION: Compared with spontaneous vaginal delivery, cesarean delivery, and operative vaginal delivery increase the risk of maternal postpartum readmission. LEVEL OF EVIDENCE: II-2. PMID- 15802415 TI - Postpartum intravenous dexamethasone for severely preeclamptic patients without hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome: a randomized trial. AB - OBJECTIVE: We compared maternal outcomes for patients with severe preeclampsia who were managed postpartum with or without adjunctive intravenous dexamethasone. METHODS: This study was a randomized, blinded placebo-controlled clinical trial comparing the use of dexamethasone postpartum (10 mg-10 mg-5 mg-5 mg intravenously every 12 hours) with a saline control in patients with severe preeclampsia. The Student t and chi(2) tests were used for data analysis, with P < .05 considered significant. RESULTS: Data from 157 patients (77 patients receiving dexamethasone, 80 patients receiving placebo) who were treated during 2000-2003 were analyzed. Demographics, diagnostic criteria, baseline laboratory values, and postpartum outcomes were similar between groups. Although dexamethasone-treated patients had fewer returns (6.5% compared with 11.3%) to the labor/delivery/recovery unit for uncontrolled hypertension than control patients, no significant differences were found in blood pressure, antihypertensive requirements, laboratory values, length of hospitalization, interval urine output at 48 hours postpartum, or major maternal morbidity. Two control patients developed hemolysis, elevated liver enzymes, low platelets syndrome. CONCLUSION: Adjunctive use of intravenous dexamethasone for postpartum patients with severe preeclampsia does not reduce disease severity or duration. LEVEL OF EVIDENCE: I. PMID- 15802417 TI - Antibiotics for bacterial vaginosis or Trichomonas vaginalis in pregnancy: a systematic review. AB - OBJECTIVE: To determine whether antibiotic treatment for bacterial vaginosis or Trichomonas vaginalis during pregnancy decreases the risk of preterm birth and associated adverse outcomes. DATA SOURCES: Pre-MEDLINE and MEDLINE (1966-2003), EMBASE (1980-2003), and the Cochrane Library were searched using the keywords "bacterial vaginosis", "Trichomonas", "Trichomonas vaginalis", "Trichomonas vaginitis", "Trichomonas infections", "pregnancy", "pregnant", "antibiotics", and "antibiotic prophylaxis". METHODS OF STUDY SELECTION: The search produced 1,888 titles, of which 1,256 abstracts were reviewed further. Of these, 1,217 were ineligible. Inclusion criteria were the following: randomized controlled trials in which antibiotics were compared with no antibiotic or placebo, for women in the second or third trimester of pregnancy with symptomatic or asymptomatic bacterial vaginosis or Trichomonas vaginalis, intact membranes, and not in labor. Exclusion criteria were as follows: published in a language other than English, dropout rate of more than 20% of women in either group, and lack of usable outcomes. Of the 39 papers reviewed in detail, 14 studies were included in the meta-analysis. TABULATION, INTEGRATION, AND RESULTS: One of the authors reviewed titles obtained from the searches, and 2 reviewers independently reviewed the abstracts, excluded those that were ineligible, identified eligible papers, and abstracted the data. For women with bacterial vaginosis, antibiotics reduced the risk of persistent infection but did not reduce the risk of preterm birth or the incidence of associated adverse outcomes for the general population or for any subgroup analyzed. For women with Trichomonas vaginalis, metronidazole reduced the risk of persistent infection but increased the incidence of preterm birth. CONCLUSION: Contrary to the conclusions of 3 recent systematic reviews, we found no evidence to support the use of antibiotic treatment for bacterial vaginosis or Trichomonas vaginalis in pregnancy to reduce the risk of preterm birth or its associated morbidities in low- or high-risk women. PMID- 15802416 TI - Effectiveness and safety of ginger in the treatment of pregnancy-induced nausea and vomiting. AB - OBJECTIVE: Conventional antiemetics are burdened with the potential of teratogenic effects during the critical embryogenic period of pregnancy. Thus, a safe and effective medication would be a welcome addition to the therapeutic repertoire. This systematic review was aimed at assessing the evidence for or against the efficacy and safety of ginger (Zingiber officinale) therapy for nausea and vomiting during pregnancy. DATA SOURCES: Systematic literature searches were conducted in 3 computerized databases (MEDLINE, EMBASE, and Cochrane Library), and the reference lists of all papers located were checked for further relevant publications. METHODS OF STUDY SELECTION: For the evaluation of efficacy, only double-blind, randomized controlled trials (RCTs) were included. All retrieved clinical data, including uncontrolled trials, case reports, observational studies, and RCTs, were included in the review of safety. TABULATION, INTEGRATION, AND RESULTS: Six double-blind RCTs with a total of 675 participants and a prospective observational cohort study (n = 187) met all inclusion criteria. The methodological quality of 4 of 5 RCTs was high. Four of the 6 RCTs (n = 246) showed superiority of ginger over placebo; the other 2 RCTs (n = 429) indicated that ginger was as effective as the reference drug (vitamin B6) in relieving the severity of nausea and vomiting episodes. The observational study retrieved and RCTs (including follow-up periods) showed the absence of significant side effects or adverse effects on pregnancy outcomes. There were no spontaneous or case reports of adverse events during ginger treatment in pregnancy. CONCLUSION: Ginger may be an effective treatment for nausea and vomiting in pregnancy. However, more observational studies, with a larger sample size, are needed to confirm the encouraging preliminary data on ginger safety. LEVEL OF EVIDENCE: I. PMID- 15802418 TI - Conservative treatment of ectopic pregnancy in a cesarean scar. AB - BACKGROUND: Pregnancy developing in a cesarean scar is a very rare but possibly life-threatening condition because of the risk of rupture and excessive hemorrhage. CASE: A 34-year-old woman presented with lower abdominal pain at 6 weeks of gestation. A cesarean delivery had been performed 3 years earlier. Transvaginal ultrasound examination revealed a viable pregnancy developing in the anterior wall of the uterus. The patient was treated successfully with systemic methotrexate and curettage. CONCLUSION: Conservative management with methotrexate and curettage can be considered in the treatment of ectopic cesarean scar pregnancy. PMID- 15802419 TI - Persistent fetal hemoglobin in maternal circulation complicating the diagnosis of fetomaternal hemorrhage. AB - BACKGROUND: Transplacental hemorrhage can be life threatening to a fetus and has important maternal treatment implications. In contrast, hereditary persistence of fetal hemoglobin is a condition that has little consequence. The Kleihauer-Betke test, which is routinely used to document transplacental hemorrhage, will be positive in either case. CASES: We report two cases in which maternal persistence of fetal hemoglobin was unknown and led to the erroneous diagnosis of fetomaternal hemorrhage. These cases highlight both the limitations of the Kleihauer-Betke test and the role of flow cytometry in diagnosing fetomaternal hemorrhage. CONCLUSION: The use of flow cytometry can clarify Kleihauer-Betke test results when there is known maternal persistence of fetal hemoglobin and can more precisely quantify a fetomaternal hemorrhage for accurate Rh immune globulin dosing. PMID- 15802420 TI - Thrombosis in a pregnant hemophilia A carrier after intrapartum recombinant factor VIII. AB - BACKGROUND: Symptomatic hemophilia A is a rare disorder in females. Pregnancy and delivery in such women can be life threatening. Obstetric management is challenging and requires a multidisciplinary approach to ensure a good outcome. CASE: A woman with hemophilia A delivered by cesarean developed a deep vein thrombosis 10 days postpartum after recombinant factor VIII administration. CONCLUSION: Hemophilia A due to skewed X-inactivation is a rare cause of peripartum bleeding. Recombinant factor VIII administration can prevent hemorrhage during and after cesarean delivery but may be associated with development of deep vein thrombosis. PMID- 15802421 TI - Predictors of leiomyoma recurrence after myomectomy. AB - OBJECTIVE: The purpose of this study was to evaluate the factors associated with the recurrence of leiomyomata after myomectomy. METHODS: One hundred forty-five consecutive cases of myomectomy by laparotomy were studied retrospectively. Leiomyoma recurrence, diagnosed by transvaginal ultrasonography, was evaluated by life-table analysis and log-rank tests according to clinical characteristics of patients. RESULTS: The 5-year cumulative rates for leiomyoma recurrence and subsequent major surgery were 62% and 9%, respectively. At 5 years, the cumulative probability of recurrence was significantly lower in patients with a single leiomyoma removed (11%), compared with patients with multiple leiomyomata (74%) (P = .011); it was also lower in patients with intraoperative uterine size 10 menstrual weeks or less (46%), compared with more than 10 menstrual weeks (82%) (P = .032). However, there was a strong association of uterine size with the number of leiomyomata removed (P = .009). Childbirth after myomectomy was associated with a lower recurrence rate; the 5-year cumulative probability of recurrence was 26% in patients with subsequent parity, compared with 76% in those without subsequent parity (P = .010). CONCLUSION: Solitary myomectomy and smaller intraoperative uterine size are associated with lower rates of leiomyoma recurrence after myomectomy; the significance of uterine size may be affected by its correlation with the number of leiomyomata removed. Subsequent parity is associated with a lower probability of recurrence, but the cause and effect relationship between these two variables is unclear. LEVEL OF EVIDENCE: II-3. PMID- 15802422 TI - The diagnosis and management of progressive dysfunction of health care organizations. AB - This paper presents an ethically justified approach to the diagnosis and management of progressive dysfunction of health care organizational cultures. We explain the concept of professional integrity in terms of the ethical concept of the cofiduciary responsibility of physicians and health care organizations. We identify the ethical features of a healthy health care organization and the spectrum of progressive dysfunction of organizational cultures from cynical through wonderland and Kafkaesque to postmodern. Physicians should respond to cynical health care organizations by creating moral enclaves of professional integrity for the main purpose of confrontation and reform, to wonderland organizations by strengthening moral enclaves for the main purpose of resisting self-deception, to Kafkaesque organizations by strengthening moral enclaves still further for the main purpose of defending professional integrity (adopting a Machiavellian appearance of virtue as necessary), and to postmodern organizations by creating moral fortresses and, should these fail, quitting. PMID- 15802423 TI - "She was, like, 6 centimeters....". PMID- 15802424 TI - Abdominal pain, bloating, and urgency. PMID- 15802425 TI - Results of the national study of vaginal birth after cesarean in birth centers. PMID- 15802426 TI - Results of the national study of vaginal birth after cesarean in birth centers. PMID- 15802428 TI - Postpartum preeclampsia management with furosemide: a randomized clinical trial. PMID- 15802429 TI - Risk factors for bladder injury during cesarean delivery. PMID- 15802432 TI - First-trimester nasal bone evaluation for aneuploidy in the general population. PMID- 15802434 TI - Trends in congenital malformations, 1974-1999: effect of prenatal diagnosis and elective termination. PMID- 15802436 TI - ACOG Practice Bulletin. Clinical Management Guidelines for Obstetrician Gynecologists. Number 61, April 2005. Human papillomavirus. AB - More than 15 years ago, a relationship between human papillomavirus (HPV) infection and cervical cancer was recognized. Since then, important strides in understanding the virus have been made, particularly in the following areas: modes of transmission and risk factors associated with transmission; the oncogenic potential of specific viral types and the mechanism by which they cause cancer; and the spectrum of infection, ranging from asymptomatic carrier states to overt warts, preneoplastic lesions, and invasive cancer. Sophisticated new tests for the detection of HPV that hold great promise for improved screening for cervical cancer precursors and invasive cancer and for the triage of abnormal cervical cytology also have been developed. Understanding the immunology of HPV has allowed the development of new and more effective treatment modalities for HPV infection and the preliminary development of primary prevention modalities, including HPV vaccines. PMID- 15802438 TI - ACOG Committee Opinion. Number 310, April 2005. Endometriosis in adolescents. AB - Historically thought of as a disease that affects adult women, endometriosis increasingly is being diagnosed in the adolescent population. This disorder, which was originally described more than a century ago, still represents a vague and perplexing entity that frequently results in chronic pelvic pain, adhesive disease, and infertility. The purpose of this Committee Opinion is to highlight the differences in adolescent and adult types of endometriosis. Early diagnosis and treatment during adolescence may decrease disease progression and prevent subsequent infertility. PMID- 15802439 TI - ACOG Committee Opinion. Number 311, April 2005. Appropriate use of laparoscopically assisted vaginal hysterectomy. AB - The technique used for hysterectomy should be dictated by the indication for the surgery, patient characteristics, and patient preference. Most patients requiring hysterectomy should be offered the vaginal approach when technically feasible and medically appropriate. If specific additional procedures that can be completed laparoscopically are anticipated before surgery, laparoscopically assisted vaginal hysterectomy may be an appropriate alternative to abdominal hysterectomy. The benefits of laparoscopically assisted vaginal hysterectomy must be weighed against the potentially increased risk and expense of two distinct operative procedures, laparoscopy and vaginal hysterectomy. PMID- 15802441 TI - Reporting standards for inferior vena caval filter placement and patient follow up: supplement for temporary and retrievable/optional filters. PMID- 15802442 TI - Incorporating CT, MR imaging, and positron emission tomography into minimally invasive therapies. PMID- 15802443 TI - Selection criteria for patients undergoing transjugular intrahepatic portosystemic shunt procedures: current status. AB - The transjugular intrahepatic portosystemic shunt (TIPS) procedure has a well established role in the management of patients with complications of portal hypertension such as variceal bleeding or refractory ascites. Several clinical variables have been described to be associated with a poor prognosis after a TIPS procedure, including the presence of uncontrollable ascites, the number of sclerotherapy sessions to control a bleeding episode, the use of drugs for hemodynamic support, the use of balloon tamponade to control bleeding, the need for an emergency TIPS procedure, the need for mechanical ventilation, prothrombin time, increased serum creatinine, increased serum bilirubin, encephalopathy, and sepsis. In addition, several scoring systems have been developed and applied to patients undergoing TIPS procedures in an attempt to improve patient selection criteria for this invasive procedure. This article reviews the most important scoring systems that have been developed and applied to patients undergoing emergency or elective TIPS procedures, with particular emphasis on the prognostic index designed for patients undergoing emergency TIPS procedures and the Model for End-stage Liver Disease score designed for patients undergoing elective TIPS procedures. The most practical application of these scoring systems is probably that, with the information provided, the operator is able to discuss with referring physicians, patients, and family members the expected outcomes of this challenging procedure. PMID- 15802444 TI - Influence of extrahepatic arterial inflow into the posterior segment or caudate lobe of the liver on repeated hepatic arterial infusion chemotherapy. AB - PURPOSE: For therapeutic effectiveness of repeated hepatic arterial infusion chemotherapy (HAIC) for unresectable advanced liver malignancies, distribution of anticancer drugs via an indwelling catheter should be uniform throughout the entire liver. It was investigated how an extrahepatic artery entering the posterior segment or caudate lobe of the liver influences performance of repeated HAIC. MATERIALS AND METHODS: One hundred ninety-five patients with unresectable advanced liver cancer underwent placement of a percutaneously implantable port catheter system and were followed arteriographically with and without computed tomography (CT). The frequency of poor distribution in the posterior segment or caudate lobe of the liver was observed on CT during arteriography via the port for HAIC. The correlation between poor distribution and various factors including the number of intrahepatic arteries was studied, and the management of such poor distribution is described. RESULTS: In 34 of 195 patients (17.4%), poor distribution was observed in the posterior segment and/or caudate lobe of the liver. The rate of poor distribution was significantly higher in those with two or more hepatic arteries than in those with one (13 of 35 vs 21 of 160; P = .0007, chi2 test). Other evaluated factors were not significantly correlated with such poor distribution. The right inferior phrenic artery was successfully embolized in 22 patients with tumors in segments with poor distribution. Good distribution throughout the entire liver was achieved in 21 patients (95.5%). CONCLUSION: Inflow of the right inferior phrenic artery into the liver occasionally prevents distribution of anticancer drugs throughout the entire liver during performance of long-term HAIC. PMID- 15802445 TI - Efficacy and safety of hepatic artery infusion catheter placement without fixation in the right gastroepiploic artery. AB - PURPOSE: To evaluate the feasibility of placing the tip of an infusion catheter in the right gastroepiploic artery via the femoral route and whether coil placement in the gastroduodenal artery and around the infusion catheter tip should be performed to avoid gastroduodenal toxicity and catheter dislocation. MATERIALS AND METHODS: Seventy-eight patients (25 women and 53 men; age, 38-79 years; mean, 63 years) underwent implantation of a 5-F chemotherapeutic infusion catheter via the femoral artery, positioned so that the tip was in the right gastroepiploic artery and a side hole was in the common hepatic artery. Patients were randomly divided into two groups: group A included patients with coil placement in the gastroduodenal artery and around the infusion catheter tip and group B included patients without coil placement. RESULTS: Catheter placement via the femoral route was successful in 70 of 78 patients (90%). Mean catheter indwell durations were 250 days (range, 0-962 days) in group A (n = 35) and 230 days (range, 0-834 days) in group B (n = 43; P = .9). Complications relating to long-term catheter duration in the right gastroepiploic artery did not occur in any patient. Two patients in each group had abdominal pain during infusion chemotherapy. Endoscopy revealed acute gastric mucosal lesions. Dislocation of the catheter tip during treatment occurred in one patient in each group. CONCLUSION: Catheter placement in the right gastroepiploic artery is safe and feasible except in patients with a long and caudally oriented celiac trunk. Coil embolization of the gastroduodenal artery and around the catheter tip is unnecessary. PMID- 15802446 TI - Catheter redundancy in the aortic arch increases the risk of stroke in left subclavian arterial port-catheter systems. AB - PURPOSE: The stroke rate after left subclavian arterial port-catheter placement was compared in two groups: one with minimal redundancy of the catheter and one with pronounced redundancy in the aortic arch designed to minimize the likelihood of catheter dislocation. MATERIALS AND METHODS: One hundred forty-eight patients (102 men, 46 women; age range, 26-83 years; mean age, 64.3 years) with inoperable advanced liver cancers underwent percutaneous implantation of port-catheter systems via the left subclavian artery. In 33 patients, a pronounced redundancy of the catheter was intentionally looped in the aortic arch. Redundancy was intentionally avoided in the remaining 115 patients. The rates of brain infarction complications in these two groups were retrospectively compared. RESULTS: Among the 33 patients with pronounced redundant catheter looping, brain infarctions occurred in four cases (12.1%). In contrast, brain infarctions occurred in only three of the 115 patients with minimal redundant catheter looping (2.6%). The frequency of brain infarction complications was significantly higher in the patients with pronounced redundant catheter looping in the aortic arch than in patients with minimal pronounced redundant catheter looping (P = .044, Fisher exact test). CONCLUSION: Catheter redundancy in the aortic arch increases the risk of stroke in patients undergoing implantation of port-catheter systems via a left subclavian arterial approach for repeated hepatic arterial infusion chemotherapy. PMID- 15802447 TI - Efficacy of manual aspiration immediately after complicated pneumothorax in CT guided lung biopsy. AB - PURPOSE: The goal of this study was to evaluate the efficacy of simple aspiration of air from the pleural space to prevent increased pneumothorax and avoid chest tube placement in cases of pneumothorax after computed tomography (CT)-guided lung biopsy. MATERIALS AND METHODS: This retrospective study was based on experience with 283 consecutive percutaneous needle lung biopsies with real-time CT fluoroscopic guidance. While patients were on the CT scanner table, percutaneous manual aspiration was performed in all those with moderate or large pneumothorax demonstrated on postbiopsy chest CT images regardless of symptoms. The authors evaluated the frequency of biopsy-induced pneumothorax, management of each such case, and factors that influenced the incidence of worsening pneumothorax that required chest tube placement despite manual aspiration. RESULTS: Of the 104 (36.7%) pneumothoraces occurring after 283 biopsy procedures, 52 were treated with manual aspiration immediately after biopsy. In 95 of the 104 pneumothoraces (91.3%), the pneumothorax had resolved completely on follow-up chest radiographs without chest tube placement. Only nine patients (3.2% of the entire series; 8.7% of those who developed pneumothorax) required chest tube placement. Requirement of chest tube insertion significantly increased parallel to the increased volume of aspirated air. The optimal cutoff level of aspirated air on which to base a decision to abandon manual aspiration alone and resort to chest tube placement was 543 mL. CONCLUSION: Percutaneous manual aspiration of biopsy-induced pneumothorax performed immediately after biopsy may prevent progressive pneumothorax and eliminate the need for chest tube placement. However, in cases in which the amount of aspirated air is large (such as more than 543 mL in this study), the possibility of required chest tube placement increases. PMID- 15802448 TI - Needle tract seeding after radiofrequency ablation of hepatic tumors. AB - PURPOSE: To determine the incidence and risk factors associated with needle tract seeding after radiofrequency ablation (RFA) of liver tumors. MATERIALS AND METHODS: A prospective data base of patients with hepatic tumors treated by RFA from December 1999 until August 2003 was reviewed to identify patients with needle tract seeding. During this period, 200 patients (148 men, 52 women) with 299 lesions underwent 298 treatment sessions. Patients with both primary (153 hepatocellular carcinoma, two cholangiocarcinoma) and a variety of secondary tumors (35 colorectal, 10 other) were treated. RFA was performed percutaneously with computed tomography (CT) and/or ultrasound (US) guidance, or with US guidance at laparoscopy or laparotomy. All procedures were performed with a LeVeen needle electrode. The needle tract was not routinely coagulated or embolized. RESULTS: Eight patients out of 200 (4%) were identified with needle tract seeding, based on imaging findings or surgical reintervention. This corresponds to a rate of eight of 298 (2.7%) per treatment session and eight of 299 (2.7%) per lesion. Statistically significant risk factors for neoplastic seeding included treatment of a subcapsular lesion (OR = 11.57, P = .007), multiple treatment sessions (OR = 2.0, P = .037), and multiple electrode placements (OR = 1.4, P = .006). CONCLUSIONS: Neoplastic seeding may occur after RFA of liver tumors. The results show that the frequency of this complication is not insignificant, and are at the upper end of rates reported in the literature of 0.5% to 2.8%. Specific risk factors identified in this study include treatment of subcapsular lesions, patients treated in multiple sessions, and lesions requiring more than one electrode placement. PMID- 15802449 TI - Navigation with electromagnetic tracking for interventional radiology procedures: a feasibility study. AB - PURPOSE: To assess the feasibility of the use of preprocedural imaging for guide wire, catheter, and needle navigation with electromagnetic tracking in phantom and animal models. MATERIALS AND METHODS: An image-guided intervention software system was developed based on open-source software components. Catheters, needles, and guide wires were constructed with small position and orientation sensors in the tips. A tetrahedral-shaped weak electromagnetic field generator was placed in proximity to an abdominal vascular phantom or three pigs on the angiography table. Preprocedural computed tomographic (CT) images of the phantom or pig were loaded into custom-developed tracking, registration, navigation, and rendering software. Devices were manipulated within the phantom or pig with guidance from the previously acquired CT scan and simultaneous real-time angiography. Navigation within positron emission tomography (PET) and magnetic resonance (MR) volumetric datasets was also performed. External and endovascular fiducials were used for registration in the phantom, and registration error and tracking error were estimated. RESULTS: The CT scan position of the devices within phantoms and pigs was accurately determined during angiography and biopsy procedures, with manageable error for some applications. Preprocedural CT depicted the anatomy in the region of the devices with real-time position updating and minimal registration error and tracking error (<5 mm). PET can also be used with this system to guide percutaneous biopsies to the most metabolically active region of a tumor. CONCLUSIONS: Previously acquired CT, MR, or PET data can be accurately codisplayed during procedures with reconstructed imaging based on the position and orientation of catheters, guide wires, or needles. Multimodality interventions are feasible by allowing the real-time updated display of previously acquired functional or morphologic imaging during angiography, biopsy, and ablation. PMID- 15802450 TI - Partition of calibrated tris-acryl gelatin microspheres in the arterial vasculature of embolized nasopharyngeal angiofibromas and paragangliomas. AB - PURPOSE: To determine the location of calibrated tris-acryl gelatin microspheres (TGMs) in the arterial vasculature of nasopharyngeal angiofibromas (NAFs) and paragangliomas (PGs). MATERIALS AND METHODS: Forty-nine specimens (25 PGs and 24 NAFs) treated operatively after embolization with TGMs of various sizes (100-300 microm to 900-1200 microm) were stained with hematoxylin and eosin saffron and analyzed at an objective magnification of 10 or 20 with a micrometric eyepiece (magnification, x12.5). The diameter of occluded vessels, their localization (intra- or extratumoral), and the number and diameter of TGMs they contained were determined. RESULTS: Embolized vessels (N = 1125) were measured: 440 in PGs and 685 in NAFs. Vessels were 89% intratumoral and 11% extratumoral. The diameter of the occluded vessels increased significantly with the size range of TGMs used for embolization for each tumor type (P < .0001). Intratumoral occluded vessels were significantly smaller than extratumoral vessels (P < .0001). Distribution of TGMs within the vascular network (intratumoral or extratumoral location) were similar for NAFs and PGs. The intratumoral and extratumoral dissemination of TGMs was different when comparing 100-300-microm TGMs versus 500-700-microm TGMs (P = .0006) as well as 300-500-microm TGMs versus 500-700-microm TGMs (P = .0001). CONCLUSIONS: The size of the vessels occluded by TGMs and their intra- or extratumoral location directly depend on the size of the injected TGMs. The vessels located inside the tumors were smaller than those located outside the tumors. A threshold for the intratumoral penetration of TGMs in the vasculature can be proposed from these data. There was no evidence of different behavior of TGMs in NAFs versus PGs. PMID- 15802451 TI - An experimental model of Stanford type B aortic dissection. AB - PURPOSE: To create an experimental model of aortic dissection with a long-lasting patent false lumen as a proper animal model for development of less-invasive treatment for aortic dissection. MATERIALS AND METHODS: Fifteen adult beagle dogs (weight, 10-12 kg) were used. The descending aorta was exposed by a left thoracotomy at the sixth intercostal space. The entry for the aortic dissection was created surgically just distal to the origin of the left innominate artery and the reentry was 5 cm distal to the entry point. Normal saline solution was injected into the aortic wall (ie, media) between these two points to create the dissection. The dogs were followed up at 1 day, 3 months, 1 year, and 2 years. RESULTS: All 12 surviving dogs had completely patent true and false lumina without any thrombi. Microscopic examination showed that the dissection was created in the tunica media layer, making it identical to aortic dissection in humans. Color Doppler imaging confirmed the patency of the true and false lumina and the relatively narrowed true lumen. CONCLUSION: In this canine model of aortic dissection, the false lumen has excellent long-term patency and the dissection plane is histologically similar to that in human aortic dissection. This model may contribute to the development of new treatments for Stanford type B aortic dissection. PMID- 15802452 TI - Metallic ureteral stents versus endoureterotomy as a therapeutic approach for experimental ureteral stricture. AB - PURPOSE: To determine if the deployment of metallic ureteral stents in benign ureteral stricture is more effective than endoureterotomy. MATERIALS AND METHODS: Twenty pigs were randomly divided in two groups to be treated with endoureterotomy or metallic stents. The internal ureteral diameter 2 cm distal to the ureteropelvic junction, histopathologic changes, ultrasonographic and fluoroscopic studies, urine culture, and serum urea and creatinine levels were analyzed during the phases of the study. The study was divided into three phases. The first included premodel documentation of normal urinary tract and laparoscopic ureteral stricture induction. In the second phase 1 month later, diagnosis and endoureteral treatment were carried out. Twelve weeks after stricture treatment, follow-up imaging studies and postmortem evaluation of all animals were performed. RESULTS: The success rates were 80% for endoureterotomy and 70% for metallic stent implantation. In all animals in the latter group, a ureteral hyperplasic reaction took place that affected the zone of stent implantation. One difference between the groups was the presence of urinary infection in 30% of animals in the endoureterotomy group and in no animals treated with metallic stents. Statistically significant differences in ureteral diameter between the first and second study phases were found within both groups (P < .0001), but not between groups (P = .021). Results from pathologic analysis revealed statistically significant differences between groups (P = .0029). CONCLUSIONS: The interaction among the distal end of the metallic stent, the urothelium, and peristalsis might be the origin of the failure of these devices. Although the deployment of metallic stents is attractive compared with other endourologic techniques like endoureterotomy in cases of benign strictures, certain problems must be resolved to prevent hyperplasia. Until then, this technique will not be a fully reliable therapeutic option for ureteral disorders. PMID- 15802453 TI - Retrieval of a malpositioned vena cava filter with embolic protection with use of a second filter. AB - Proper positioning of inferior vena cava (IVC) filters is necessary for effective protection from pulmonary embolism (PE). This report describes a case of an IVC filter malpositioned partially in the right common iliac vein that resulted in risk of recurrent PE caused by thrombus emanating from the left common iliac vein. A new filter was placed in the IVC above the first device, followed by retrieval of the malpositioned filter through the new filter. PMID- 15802454 TI - Aortic pseudoaneurysm after penetration by a Simon nitinol inferior vena cava filter. AB - This report describes an unusual complication related to inferior vena cava (IVC) filter placement. A 50-year-old woman undergoing long-term anticoagulation presented to her primary care physician with abdominal pain after a motor vehicle accident. An IVC filter had been placed 7 years earlier. Computed tomography of the abdomen demonstrated a moderate perisplenic hematoma and a fragmented IVC filter penetrating the aorta. A small infrarenal aortic pseudoaneurysm had developed at the penetration site. Wallgraft placement successfully sealed the aneurysm. PMID- 15802455 TI - Aspiration thrombectomy of acute complete carotid bulb occlusion. AB - Two patients with acute complete carotid bulb occlusion were treated using intra arterial aspiration thrombectomy a mean of 3 hours after stroke onset. A slightly angulated 8-F guiding catheter designed for general interventional use was employed to aspirate the thrombus in the carotid artery. Manual aspiration through a 50-mL syringe resulted in effective removal of the thrombus followed by good patency of the internal carotid artery. Aspiration thrombectomy is a simple, feasible and effective rescue procedure for patients with acute complete carotid bulb occlusion. PMID- 15802456 TI - Benign duodenal strictures: treatment by means of fluoroscopically guided balloon dilation. AB - Fluoroscopically guided balloon (15 or 20 mm in diameter) dilation was performed on eight patients with benign duodenal strictures caused by peptic ulcers (n = 6), Crohn's disease (n = 1), and postoperative adhesion (n = 1). The procedure was technically and clinically successful without complications in seven of the eight patients (88%). Duodenal perforation occurred immediately after 20-mm diameter balloon dilation in one patient who underwent emergency surgery. During the mean follow-up of 30 months (range, 2-103 months), there was recurrence in two of the seven patients (29%) who then underwent surgery. The other five patients (71%) showed good results with no recurrence. PMID- 15802457 TI - Radiofrequency ablation therapy of remnant colorectal liver metastases after a course of hepatic arterial infusion chemotherapy. AB - The present study evaluated the feasibility of combined therapy employing repeated hepatic arterial infusion chemotherapy (HAIC) via a port-catheter system and radiofrequency (RF) ablation for unresectable metastatic liver cancer from the colo-rectum. RF ablation was performed for six patients with liver malignancies that had been well controlled and had decreased in number and size with repeated HAIC. After RF ablation subsequent to repeated HAIC, all six patients had stable disease or complete or partial remission as documented by follow-up contrast-enhanced computed tomographic (CT) imaging. Fluorodeoxyglucose positron emission tomography (18F) showed complete response in all ablated lesions. In conclusion, RF ablation after a course of HAIC achieved complete necrosis in residual lesions after HAIC therapy. PMID- 15802458 TI - Residual thrombus within a retrievable IVC filter. PMID- 15802459 TI - Adverse cardiac events as a result of high volume contrast injection during rotational arch aortography. PMID- 15802460 TI - Percutaneous needle biopsy under CT fluoroscopic guidance for cardiac tumor during continuous intravenous injection of contrast material. PMID- 15802461 TI - The emperor's new clothes. PMID- 15802462 TI - A case of posttraumatic twiddler's syndrome. PMID- 15802464 TI - Re: Cutaneous thermal injury after endovenous laser ablation of the great saphenous vein. PMID- 15802466 TI - Differential shuffling of native genetic diversity across introduced regions in a brown alga: aquaculture vs. maritime traffic effects. AB - Worldwide marine invaders, such as the brown alga Undaria pinnatifida, offer challenging models for unraveling the apparent paradox of sustainable settlement of exotic species over a large spectrum of environments. Two intergenic noncoding mitochondrial loci were found to be highly informative at the within-species level. Twenty-five haplotypes were found over the whole dataset (333 base pairs, 524 individuals, and 24 populations). The native range showed striking population genetic structure stemming from low diversity within and high differentiation among populations, a pattern not observed in the introduced range of this seaweed. Contrary to classical expectations of founding effects associated with accidental introduction of exotic species, most of the introduced populations showed high genetic diversity. At the regional scale, genetic diversity and sequence divergence showed contrasting patterns in the two main areas of introduction (Europe and Australasia), suggesting different processes of introduction in the two regions. Gene genealogy analyses point to aquaculture as a major vector of introduction and spread in Europe but implicate maritime traffic in promoting recurrent migration events from the native range to Australasia. The multiplicity of processes and genetic signatures associated with the successful invasion confirms that multiple facets of global change, e.g., aquaculture practices, alteration of habitats, and increased traffic, act in synergy at the worldwide level, facilitating successful pandemic introductions. PMID- 15802467 TI - Symmetrical base preferences surrounding HIV-1, avian sarcoma/leukosis virus, and murine leukemia virus integration sites. AB - To investigate retroviral integration targeting on a nucleotide scale, we examined the base frequencies directly surrounding cloned in vivo HIV-1, murine leukemia virus, and avian sarcoma/leukosis virus integrations. Base preferences of up to 2-fold the expected frequencies were found for three viruses, representing P values down to <10(-100) and defining what appear to be preferred integration sequences. Offset symmetry reflecting the topology of the integration reaction was found for HIV-1 and avian sarcoma/leukosis virus but not murine leukemia virus, suggesting fundamental differences in the way different retroviral integration complexes interact with host-cell DNA. PMID- 15802468 TI - Interaction strength combinations and the overfishing of a marine food web. AB - The stability of ecological communities largely depends on the strength of interactions between predators and their prey. Here we show that these interaction strengths are structured nonrandomly in a large Caribbean marine food web. Specifically, the cooccurrence of strong interactions on two consecutive levels of food chains occurs less frequently than expected by chance. Even when they occur, these strongly interacting chains are accompanied by strong omnivory more often than expected by chance. By using a food web model, we show that these interaction strength combinations reduce the likelihood of trophic cascades after the overfishing of top predators. However, fishing selectively removes predators that are overrepresented in strongly interacting chains. Hence, the potential for strong community-wide effects remains a threat. PMID- 15802469 TI - Independent perceptual learning in monocular and binocular motion systems. AB - Eye-transfer tests, external noise manipulations, and observer models were used to systematically characterize learning mechanisms in judging motion direction of moving objects in visual periphery (Experiment 1) and fovea (Experiment 2) and to investigate the degree of transfer of the learning mechanisms from trained to untrained eyes. Perceptual learning in one eye was measured over 10 practice sessions. Subsequent learning in the untrained eye was assessed in five transfer sessions. We characterized the magnitude of transfer of each learning mechanism to the untrained eye by separately analyzing the magnitude of subsequent learning in low and high external noise conditions. In both experiments, we found that learning in the trained eye reduced contrast thresholds uniformly across all of the external noise levels: 47 +/- 10% and 62 +/- 8% in experiments 1 and 2, respectively. Two mechanisms, stimulus enhancement and template retuning, accounted for the observed performance improvements. The degree of transfer to the untrained eye depended on the amount of external noise added to the signal stimuli: In high external noise conditions, learning transferred completely to the untrained eye in both experiments. In low external noise conditions, there was only partial transfer of learning: 63% in experiment 1 and 54% in experiment 2. The results suggest that template retuning, which is effective in high external noise conditions, is mostly binocular, whereas stimulus enhancement, which is effective in low external noise displays, is largely monocular. The two independent mechanisms underlie perceptual learning of motion direction identification in monocular and binocular motion systems. PMID- 15802470 TI - Multipotent nestin-positive, keratin-negative hair-follicle bulge stem cells can form neurons. AB - We have recently shown that the expression of nestin, the neural stem cell marker protein, is expressed in bulge-area stem cells of the hair follicle. We used transgenic mice with GFP expression driven by the nestin regulatory element [nestin-driven GFP (ND-GFP)]. The ND-GFP stem cells give rise to the outer-root sheath of the hair follicle as well as an ND-GFP interfollicular vascular network. In this study, we demonstrate that ND-GFP stem cells isolated from the hair-follicle bulge area that are negative for the keratinocyte marker keratin 15 can differentiate into neurons, glia, keratinocytes, smooth muscle cells, and melanocytes in vitro. These pluripotent ND-GFP stem cells are positive for the stem cell marker CD34, as well as keratin 15-negative, suggesting their relatively undifferentiated state. The apparent primitive state of the ND-GFP stem cells is compatible with their pluripotency. Furthermore, we show that cells derived from ND-GFP stem cells can differentiate into neurons after transplantation to the subcutis of nude mice. These results suggest that hair follicle bulge-area ND-GFP stem cells may provide an accessible, autologous source of undifferentiated multipotent stem cells for therapeutic application. PMID- 15802471 TI - The phasevarion: a genetic system controlling coordinated, random switching of expression of multiple genes. AB - Several host-adapted bacterial pathogens contain methyltransferases associated with type III restriction-modification (R-M) systems that are subject to reversible, high-frequency on/off switching of expression (phase variation). To investigate the role of phase-variable expression of R-M systems, we made a mutant strain lacking the methyltransferase (mod) associated with a type III R-M system of Haemophilus influenzae and analyzed its phenotype. By microarray analysis, we identified a number of genes that were either up- or down-regulated in the mod mutant strain. This system reports the coordinated random switching of a set of genes in a bacterial pathogen and may represent a widely used mechanism. PMID- 15802472 TI - The majority of human genes have regions repeated in other human genes. AB - Amino acid sequence comparisons have been made between all of 25,193 human proteins with each of the others by using blast software (National Center for Biotechnology Information) and recording the results for regions that are significantly related in sequence, that is, have an expectation of <1 x 10(-3). The results are presented for each amino acid as the number of identical or similar amino acids matched in these aligned regions. This approach avoids summing or dealing directly with the different regions of any one protein that are often related to different numbers and types of other proteins. The results are presented graphically for a sample of 140 proteins. Relationships are not observed for 26.5% of the 12,728,866 amino acids. The average number of related amino acids is 36.5 for the majority (73.5%) that show relationships. The median number of recognized relationships is approximately 3 for all of the amino acids, and the maximum number is 718. The results demonstrate the overwhelming importance of gene regional duplication forming families of proteins with related domains and show the variety of the resulting patterns of relationship. The magnitude of the set of relationships leads to the conclusion that the principal process by which new gene functions arise has been by making use of preexisting genes. PMID- 15802473 TI - Intermediates in the oxygenation of a nonheme diiron(II) complex, including the first evidence for a bound superoxo species. AB - The reaction of [Fe(2)(mu-OH)(2)(6-Me(3)-TPA)(2)](2+) (1) [6-Me(3)-TPA, Tris(6 methyl-2-pyridylmethyl)amine] with O(2) in CH(2)Cl(2) at -80 degrees C gives rise to two new intermediates, 2 and 3, before the formation of previously characterized [Fe(2)(O)(O(2))(6-Me(3)-TPA)(2)](2+) (4) that allow the oxygenation reaction to be monitored one electron-transfer step at a time. Raman evidence assigns 2 and 3 as a diiron-superoxo species and a diiron-peroxo species, respectively. Intermediate 2 exhibits its nu(O-O) at 1,310 cm(-1) with a -71-cm( 1) (18)O isotope shift. A doublet peak pattern for the (16)O(18)O isotopomer of 2 in mixed-isotope Raman experiments strongly suggests that the superoxide ligand of 2 is bound end-on. This first example of a nonheme iron-superoxo intermediate exhibits the highest frequency nu(O-O) yet observed for a biomimetic metal dioxygen adduct. The bound superoxide of 2, unlike the bound peroxide of 4, is readily reduced by 2,4-di-tert-butylphenol via a proton-coupled electron-transfer mechanism, emphasizing that metal-superoxo species may serve as oxidants in oxygen activation mechanisms of metalloenzymes. The discovery of intermediates 2 and 3 allows us to dissect the initial steps of dioxygen binding at a diiron center leading to its activation for substrate oxidation. PMID- 15802474 TI - Clinical decision support in electronic prescribing: recommendations and an action plan: report of the joint clinical decision support workgroup. AB - Clinical decision support (CDS) in electronic prescribing (eRx) systems can improve the safety, quality, efficiency, and cost-effectiveness of care. However, at present, these potential benefits have not been fully realized. In this consensus white paper, we set forth recommendations and action plans in three critical domains: (1) advances in system capabilities, including basic and advanced sets of CDS interventions and knowledge, supporting database elements, operational features to improve usability and measure performance, and management and governance structures; (2) uniform standards, vocabularies, and centralized knowledge structures and services that could reduce rework by vendors and care providers, improve dissemination of well-constructed CDS interventions, promote generally applicable research in CDS methods, and accelerate the movement of new medical knowledge from research to practice; and (3) appropriate financial and legal incentives to promote adoption. PMID- 15802475 TI - Automated detection of adverse events using natural language processing of discharge summaries. AB - OBJECTIVE: To determine whether natural language processing (NLP) can effectively detect adverse events defined in the New York Patient Occurrence Reporting and Tracking System (NYPORTS) using discharge summaries. DESIGN: An adverse event detection system for discharge summaries using the NLP system MedLEE was constructed to identify 45 NYPORTS event types. The system was first applied to a random sample of 1,000 manually reviewed charts. The system then processed all inpatient cases with electronic discharge summaries for two years. All system identified events were reviewed, and performance was compared with traditional reporting. MEASUREMENTS: System sensitivity, specificity, and predictive value, with manual review serving as the gold standard. RESULTS: The system correctly identified 16 of 65 events in 1,000 charts. Of 57,452 total electronic discharge summaries, the system identified 1,590 events in 1,461 cases, and manual review verified 704 events in 652 cases, resulting in an overall sensitivity of 0.28 (95% confidence interval [CI]: 0.17-0.42), specificity of 0.985 (CI: 0.984 0.986), and positive predictive value of 0.45 (CI: 0.42-0.47) for detecting cases with events and an average specificity of 0.9996 (CI: 0.9996-0.9997) per event type. Traditional event reporting detected 322 events during the period (sensitivity 0.09), of which the system identified 110 as well as 594 additional events missed by traditional methods. CONCLUSION: NLP is an effective technique for detecting a broad range of adverse events in text documents and outperformed traditional and previous automated adverse event detection methods. PMID- 15802476 TI - Optimal search strategies for detecting clinically sound prognostic studies in EMBASE: an analytic survey. AB - BACKGROUND: Clinical end users of EMBASE have a difficult time retrieving articles that are both scientifically sound and directly relevant to clinical practice. Search filters have been developed to assist end users in increasing the success of their searches. Many filters have been developed for the literature on therapy and reviews for use in MEDLINE, but little has been done for use in EMBASE with no filter development for studies of prognosis. The objective of this study was to determine how well various methodologic textwords, index terms, and their Boolean combinations retrieve methodologically sound literature on the prognosis of health disorders in EMBASE. METHODS: An analytic survey was conducted, comparing hand searches of 55 journals with retrievals from EMBASE for 4,843 candidate search terms and 8,919 combinations. All articles were rated using purpose and quality indicators, and clinically relevant prognostic articles were categorized as "pass" or "fail" according to explicit criteria for scientific merit. Candidate search strategies were run in EMBASE, the retrievals being compared with the hand search data. The sensitivity, specificity, precision, and accuracy of the search strategies were calculated. RESULTS: Of the 1,064 articles about prognosis, 148 (13.9%) met basic criteria for scientific merit. Combinations of search terms reached peak sensitivities of 98.7% with specificity at 50.6%. Compared with best single terms, best multiple terms increased sensitivity for sound studies by 12.2% (absolute increase), while decreasing specificity (absolute decrease 5.1%) when sensitivity was maximized. Combinations of search terms reached peak specificities of 93.4% with sensitivity at 50.7%. Compared with best single terms, best multiple terms increased specificity for sound studies by 7.1% (absolute increase), while decreasing sensitivity (absolute decrease 8.8%) when specificity was maximized. CONCLUSION: Empirically derived search strategies combining indexing terms and textwords can achieve high sensitivity or specificity for retrieving sound prognostic studies from EMBASE. PMID- 15802477 TI - KAT: a flexible XML-based knowledge authoring environment. AB - As part of an enterprise effort to develop new clinical information systems at Intermountain Health Care, the authors have built a knowledge authoring tool that facilitates the development and refinement of medical knowledge content. At present, users of the application can compose order sets and an assortment of other structured clinical knowledge documents based on XML schemas. The flexible nature of the application allows the immediate authoring of new types of documents once an appropriate XML schema and accompanying Web form have been developed and stored in a shared repository. The need for a knowledge acquisition tool stems largely from the desire for medical practitioners to be able to write their own content for use within clinical applications. We hypothesize that medical knowledge content for clinical use can be successfully created and maintained through XML-based document frameworks containing structured and coded knowledge. PMID- 15802478 TI - Handheld computer-based decision support reduces patient length of stay and antibiotic prescribing in critical care. AB - OBJECTIVE: This study assessed the effect of a handheld computer-based decision support system (DSS) on antibiotic use and patient outcomes in a critical care unit. DESIGN: A DSS containing four types of evidence (patient microbiology reports, local antibiotic guidelines, unit-specific antibiotic susceptibility data for common bacterial pathogens, and a clinical pulmonary infection score calculator) was developed and implemented on a handheld computer for use in the intensive care unit at a tertiary referral hospital. System impact was assessed in a prospective "before/after" cohort trial lasting 12 months. Outcome measures were defined daily doses (DDDs) of antibiotics per 1,000 patient-days, patient length of stay, and mortality. RESULTS: The number of admissions, APACHE (Acute Physiology, Age, and Chronic Health Evaluation) II and SAPS (Simplified Acute Physiology Score) II for patients in preintervention, and intervention (DSS use) periods were statistically comparable. The mean patient length of stay and the use of antibiotics in the unit during six months of the DSS use decreased from 7.15 to 6.22 bed-days (p = 0.02) and from 1,767 DDD to 1,458 DDD per 1,000 patient-days (p = 0.04), respectively, with no change in mortality. The DSS was accessed 674 times during 168 days of the trial. Microbiology reports and antibiotic guidelines were the two most commonly used (53% and 22.5%, respectively) types of evidence. The greatest reduction was observed in the use of beta-lactamase-resistant penicillins and vancomycin. CONCLUSION: Handheld computer-based decision support contributed to a significant reduction in patient length of stay and antibiotic prescribing in a critical care unit. PMID- 15802479 TI - A randomized trial of electronic clinical reminders to improve quality of care for diabetes and coronary artery disease. AB - OBJECTIVE: The aim of this study was to evaluate the impact of an integrated patient-specific electronic clinical reminder system on diabetes and coronary artery disease (CAD) care and to assess physician attitudes toward this reminder system. DESIGN: We enrolled 194 primary care physicians caring for 4549 patients with diabetes and 2199 patients with CAD at 20 ambulatory clinics. Clinics were randomized so that physicians received either evidence-based electronic reminders within their patients' electronic medical record or usual care. There were five reminders for diabetes care and four reminders for CAD care. MEASUREMENTS: The primary outcome was receipt of recommended care for diabetes and CAD. We created a summary outcome to assess the odds of increased compliance with overall diabetes care (based on five measures) and overall CAD care (based on four measures). We surveyed physicians to assess attitudes toward the reminder system. RESULTS: Baseline adherence rates to all quality measures were low. While electronic reminders increased the odds of recommended diabetes care (odds ratio [OR] 1.30, 95% confidence interval [CI] 1.01-1.67) and CAD (OR 1.25, 95% CI 1.01 1.55), the impact of individual reminders was variable. A total of three of nine reminders effectively increased rates of recommended care for diabetes or CAD. The majority of physicians (76%) thought that reminders improved quality of care. CONCLUSION: An integrated electronic reminder system resulted in variable improvement in care for diabetes and CAD. These improvements were often limited and quality gaps persist. PMID- 15802480 TI - Toward semantic interoperability in home health care: formally representing OASIS items for integration into a concept-oriented terminology. AB - OBJECTIVE: The authors aimed to (1) formally represent OASIS-B1 concepts using the Logical Observation Identifiers, Names, and Codes (LOINC) semantic structure; (2) demonstrate integration of OASIS-B1 concepts into a concept-oriented terminology, the Medical Entities Dictionary (MED); (3) examine potential hierarchical structures within LOINC among OASIS-B1 and other nursing terms; and (4) illustrate a Web-based implementation for OASIS-B1 data entry using Dialogix, a software tool with a set of functions that supports complex data entry. DESIGN AND MEASUREMENTS: Two hundred nine OASIS-B1 items were dissected into the six elements of the LOINC semantic structure and then integrated into the MED hierarchy. Each OASIS-B1 term was matched to LOINC-coded nursing terms, Home Health Care Classification, the Omaha System, and the Sign and Symptom Check-List for Persons with HIV, and the extent of the match was judged based on a scale of 0 (no match) to 4 (exact match). OASIS-B1 terms were implemented as a Web-based survey using Dialogix. RESULTS: Of 209 terms, 204 were successfully dissected into the elements of the LOINC semantics structure and integrated into the MED with minor revisions of MED semantics. One hundred fifty-one OASIS-B1 terms were mapped to one or more of the LOINC-coded nursing terms. CONCLUSION: The LOINC semantic structure offers a standard way to add home health care data to a comprehensive patient record to facilitate data sharing for monitoring outcomes across sites and to further terminology management, decision support, and accurate information retrieval for evidence-based practice. The cross-mapping results support the possibility of a hierarchical structure of the OASIS-B1 concepts within nursing terminologies in the LOINC database. PMID- 15802481 TI - Implementing a commercial rule base as a medication order safety net. AB - A commercial rule base (Cerner Multum) was used to identify medication orders exceeding recommended dosage limits at five hospitals within BJC HealthCare, an integrated health care system. During initial testing, clinical pharmacists determined that there was an excessive number of nuisance and clinically insignificant alerts, with an overall alert rate of 9.2%. A method for customizing the commercial rule base was implemented to increase rule specificity for problematic rules. The system was subsequently deployed at two facilities and achieved alert rates of less than 1%. Pharmacists screened these alerts and contacted ordering physicians in 21% of cases. Physicians made therapeutic changes in response to 38% of alerts presented to them. By applying simple techniques to customize rules, commercial rule bases can be used to rapidly deploy a safety net to screen drug orders for excessive dosages, while preserving the rule architecture for later implementations of more finely tuned clinical decision support. PMID- 15802482 TI - Exploring barriers and facilitators to the use of computerized clinical reminders. AB - OBJECTIVE: Evidence-based practices in preventive care and chronic disease management are inconsistently implemented. Computerized clinical reminders (CRs) can improve compliance with these practices in outpatient settings. However, since clinician adherence to CR recommendations is quite variable and declines over time, we conducted observations to determine barriers and facilitators to the effective use of CRs. DESIGN: We conducted an observational study of nurses and providers interacting with CRs in outpatient primary care clinics for two days in each of four geographically distributed Veterans Administration (VA) medical centers. MEASUREMENTS: Three observers recorded interactions of 35 nurses and 55 physicians and mid-level practitioners with the CRs, which function as part of an electronic medical record. Field notes were typed, coded in a spreadsheet, and then sorted into logical categories. We then integrated findings across observations into meaningful patterns and abstracted the data into themes, such as recurrent strategies. Several of these themes translated directly to barriers and facilitators to effective CR use. RESULTS: Optimally using the CR system for its intended purpose was impeded by (1) lack of coordination between nurses and providers; (2) using the reminders while not with the patient, impairing data acquisition and/or implementation of recommended actions; (3) workload; (4) lack of CR flexibility; and (5) poor interface usability. Facilitators included (1) limiting the number of reminders at a site; (2) strategic location of the computer workstations; (3) integration of reminders into workflow; and (4) the ability to document system problems and receive prompt administrator feedback. CONCLUSION: We identified barriers that might explain some of the variability in the use of CRs. Although these barriers may be difficult to overcome, some strategies may increase user acceptance and therefore the effectiveness of the CRs. These include explicitly assigning responsibility for each CR to nurses or providers, improving visibility of positive results from CRs in the electronic medical record, creating a feedback mechanism about CR use, and limiting the overall number of CRs. PMID- 15802483 TI - Integrating SNOMED CT into the UMLS: an exploration of different views of synonymy and quality of editing. AB - OBJECTIVE: The integration of SNOMED CT into the Unified Medical Language System (UMLS) involved the alignment of two views of synonymy that were different because the two vocabulary systems have different intended purposes and editing principles. The UMLS is organized according to one view of synonymy, but its structure also represents all the individual views of synonymy present in its source vocabularies. Despite progress in knowledge-based automation of development and maintenance of vocabularies, manual curation is still the main method of determining synonymy. The aim of this study was to investigate the quality of human judgment of synonymy. DESIGN: Sixty pairs of potentially controversial SNOMED CT synonyms were reviewed by 11 domain vocabulary experts (six UMLS editors and five noneditors), and scores were assigned according to the degree of synonymy. MEASUREMENTS: The synonymy scores of each subject were compared to the gold standard (the overall mean synonymy score of all subjects) to assess accuracy. Agreement between UMLS editors and noneditors was measured by comparing the mean synonymy scores of editors to noneditors. RESULTS: Average accuracy was 71% for UMLS editors and 75% for noneditors (difference not statistically significant). Mean scores of editors and noneditors showed significant positive correlation (Spearman's rank correlation coefficient 0.654, two-tailed p < 0.01) with a concurrence rate of 75% and an interrater agreement kappa of 0.43. CONCLUSION: The accuracy in the judgment of synonymy was comparable for UMLS editors and nonediting domain experts. There was reasonable agreement between the two groups. PMID- 15802485 TI - Comprehensive analysis of a medication dosing error related to CPOE. AB - This case study of a serious medication error demonstrates the necessity of a comprehensive methodology for the analysis of failures in interaction between humans and information systems. The authors used a novel approach to analyze a dosing error related to computer-based ordering of potassium chloride (KCl). The method included a chronological reconstruction of events and their interdependencies from provider order entry usage logs, semistructured interviews with involved clinicians, and interface usability inspection of the ordering system. Information collected from all sources was compared and evaluated to understand how the error evolved and propagated through the system. In this case, the error was the product of faults in interaction among human and system agents that methods limited in scope to their distinct analytical domains would not identify. The authors characterized errors in several converging aspects of the drug ordering process: confusing on-screen laboratory results review, system usability difficulties, user training problems, and suboptimal clinical system safeguards that all contributed to a serious dosing error. The results of the authors' analysis were used to formulate specific recommendations for interface layout and functionality modifications, suggest new user alerts, propose changes to user training, and address error-prone steps of the KCl ordering process to reduce the risk of future medication dosing errors. PMID- 15802484 TI - Health information technology and physician-patient interactions: impact of computers on communication during outpatient primary care visits. AB - OBJECTIVE: The aim of this study was to evaluate the impact of introducing health information technology (HIT) on physician-patient interactions during outpatient visits. DESIGN: This was a longitudinal pre-post study: two months before and one and seven months after introduction of examination room computers. Patient questionnaires (n = 313) after primary care visits with physicians (n = 8) within an integrated delivery system. There were three patient satisfaction domains: (1) satisfaction with visit components, (2) comprehension of the visit, and (3) perceptions of the physician's use of the computer. RESULTS: Patients reported that physicians used computers in 82.3% of visits. Compared with baseline, overall patient satisfaction with visits increased seven months after the introduction of computers (odds ratio [OR] = 1.50; 95% confidence interval [CI]: 1.01-2.22), as did satisfaction with physicians' familiarity with patients (OR = 1.60, 95% CI: 1.01-2.52), communication about medical issues (OR = 1.61; 95% CI: 1.05-2.47), and comprehension of decisions made during the visit (OR = 1.63; 95% CI: 1.06-2.50). In contrast, there were no significant changes in patient satisfaction with comprehension of self-care responsibilities, communication about psychosocial issues, or available visit time. Seven months post introduction, patients were more likely to report that the computer helped the visit run in a more timely manner (OR = 1.76; 95% CI: 1.28-2.42) compared with the first month after introduction. There were no other significant changes in patient perceptions of the computer use over time. CONCLUSION: The examination room computers appeared to have positive effects on physician-patient interactions related to medical communication without significant negative effects on other areas such as time available for patient concerns. Further study is needed to better understand HIT use during outpatient visits. PMID- 15802486 TI - Detection and prevention of medication errors using real-time bedside nurse charting. AB - OBJECTIVE: Charting systems with decision support have been developed to assist with medication charting, but many of the features of these programs are not properly used in their clinical application. An analysis of medication error reports at LDS Hospital revealed the occurrence of errors that should have been detected and prevented by decision support features if real-time entry at the bedside had taken place. The aim of this study was to increase the real-time bedside charting behavior of nurses. DESIGN: A quasiexperimental before and after design was used. The study took place in two 40-bed surgical units, one of which served as the study unit, the other as control unit. The study unit received educational intervention about error avoidance through real-time bedside charting, and 12 weeks of monitoring and performance feedback. The real-time and bedside charting rates for the study and control units were measured before and after the intervention. RESULTS: Before the intervention on the study unit, the real-time charting rate was 59% and the bedside rate was 40%. At the conclusion of a 12-week intervention period, the real-time rate increased to 73% and the bedside rate increased to 63%. Postintervention real-time rates were 75% after eight weeks and remained at 75% after one year. Equivalent control unit real-time rates varied from 53% to 57%, and bedside rates varied from 34% to 44% during the same intervals. CONCLUSION: Targeted educational intervention and monitored feedback yielded measurable improvements in the effective use of the computerized medication charting system and must be an ongoing process. PMID- 15802488 TI - The role of beta-adrenergic receptor signaling in cardioprotection. AB - This study examines the role of the beta2-adrenergic receptor (beta2-AR) in cardioprotection. The beta2-AR couples to Gs and Gi proteins. Gs activates PKA, which phosphorylates the receptor and switches beta2-AR coupling from Gs to Gi. Prior to 20 min of global ischemia, mouse hearts were either perfused for 30 min without treatment (control), treated with 10 nmol/L of isoproterenol (ISO) for 5 min followed by 5 min washout, or preconditioned with 4 cycles of 5 min ischemia and 5 min reflow (PC). Recovery of left ventricular developed pressure (LVDP) and infarct size were measured. Intermittent ISO treatment improved post-ischemic recovery of LVDP (58.5+/-4.8% vs. 22.0+/-6.3% in control) and reduced infarct size (31.0+/-2.4% vs. 53.0+/-4.6% in control). The Gi inhibitor pertussis toxin blocked the ISO-induced improvement in postischemic LVDP and infarct size. To test the role of beta2-AR in PC, we studied mice lacking beta2-AR (beta2-AR-/-) and found that PC had no effect on postischemic LVDP or infarct size in beta2-AR /-. To test whether PKA is required for the PC and ISO-induced protection, hearts were treated with the PKA inhibitors PKI and H-89. We found that PKI and H-89 blocked the PC- and ISO-induced improvement in postischemic LVDP and infarct size. These data show an important role for beta2-AR in cardioprotection and support the novel hypothesis that preconditioning involves switching of beta2-AR coupling from Gs to Gi. PMID- 15802487 TI - Effect of CPOE user interface design on user-initiated access to educational and patient information during clinical care. AB - OBJECTIVE: Authors evaluated whether displaying context sensitive links to infrequently accessed educational materials and patient information via the user interface of an inpatient computerized care provider order entry (CPOE) system would affect access rates to the materials. DESIGN: The CPOE of Vanderbilt University Hospital (VUH) included "baseline" clinical decision support advice for safety and quality. Authors augmented this with seven new primarily educational decision support features. A prospective, randomized, controlled trial compared clinicians' utilization rates for the new materials via two interfaces. Control subjects could access study-related decision support from a menu in the standard CPOE interface. Intervention subjects received active notification when study-related decision support was available through context sensitive, visibly highlighted, selectable hyperlinks. MEASUREMENTS: Rates of opportunities to access and utilization of study-related decision support materials from April 1999 through March 2000 on seven VUH Internal Medicine wards. RESULTS: During 4,466 intervention subject-days, there were 240,504 (53.9/subject-day) opportunities for study-related decision support, while during 3,397 control subject-days, there were 178,235 (52.5/subject-day) opportunities for such decision support, respectively (p = 0.11). Individual intervention subjects accessed the decision support features at least once on 3.8% of subject days logged on (278 responses); controls accessed it at least once on 0.6% of subject-days (18 responses), with a response rate ratio adjusted for decision support frequency of 9.17 (95% confidence interval 4.6-18, p < 0.0005). On average, intervention subjects accessed study-related decision support materials once every 16 days individually and once every 1.26 days in aggregate. CONCLUSION: Highlighting availability of context-sensitive educational materials and patient information through visible hyperlinks significantly increased utilization rates for study-related decision support when compared to "standard" VUH CPOE methods, although absolute response rates were low. PMID- 15802489 TI - Different effects of angiopoietin-2 in different vascular beds: new vessels are most sensitive. AB - In this study, we used double transgenic mice with inducible expression of angiopoietin-2 (Ang2) to investigate the role of Ang2 in the retinal and choroidal circulations and in three models of ocular neovascularization (NV). Mice with induced expression of Ang2 ubiquitously, or specifically in the retina, survived and appeared grossly normal. They also had normal-appearing retinal and choroidal circulations, demonstrating that high levels of Ang2 did not induce regression of mature retinal or choroidal vessels. When Ang2 expression was induced soon after birth, there was increased density of the deep capillary bed on postnatal day (P) 11 that returned to normal by P18, the time that retinal vascular development is usually completed. In mice with ischemic retinopathy, induction of Ang2 during the ischemic period resulted in a significant increase in retinal NV, but induction of Ang2 at a later time point when ischemia (and vascular endothelial growth factor [VEGF]) was less, hastened regression of NV. In triple transgenic mice that coexpressed VEGF and Ang2, the increased expression of Ang2 inhibited VEGF-induced NV in the retina. Increased expression of Ang2 also resulted in regression of choroidal neovascularization. These data suggest that ocular neovascularization, but not mature retinal or choroidal vessels, is sensitive to Ang2; a high Ang2/VEGF ratio promotes regression, while high Ang2 in the setting of hypoxia and/or concomitantly high Ang2 and VEGF stimulate neovascularization. PMID- 15802490 TI - The anti-psoriatic drug anthralin accumulates in keratinocyte mitochondria, dissipates mitochondrial membrane potential, and induces apoptosis through a pathway dependent on respiratory competent mitochondria. AB - Anthralin is a potent topical drug, inducing clearance of psoriatic plaques. Anthralin disrupts mitochondrial function and structure, but its mechanism of action remains undefined. This study aimed to determine whether anthralin induced keratinocyte apoptosis as well as to investigate molecular mechanisms and the role of mitochondria. We studied human keratinocytes and human 143B rho(0) cells, which lack mitochondrial DNA and a functional respiratory chain. We show that anthralin disrupts mitochondrial membrane potential (DeltaPsim) and causes endogenous cytochrome c release, resulting in the activation of caspase-3 and characteristic morphological changes of apoptosis. Disruption of DeltaPsim and cytochrome c release were independent of mitochondrial permeability transition or caspase activation. Human 143B rho(0) cells were resistant to anthralin-induced cell death, disruption of DeltaPsim, and cytochrome c release compared with the isogenic 143B rho+ cell line. Using the intrinsic fluorescence of anthralin, rapid accumulation within mitochondria was observed independent of DeltaPsim. Using assays that measure individual respiratory chain complexes, we show that anthralin specifically interacts with ubiquinone pool. These data indicate that anthralin induces apoptosis through a novel mitochondrial pathway dependent on oxidative respiration and involving electron transfer with the ubiquinone pool. These studies identify keratinocyte apoptosis as a potentially important mechanism involved in the clearance of psoriasis. PMID- 15802491 TI - Somatic DNA damage in interventional cardiologists: a case-control study. AB - Interventional cardiologists who work in cardiac catheterization laboratories are exposed to low doses of ionizing radiation that could pose a health hazard. DNA damage is considered to be the main initiating event by which radiation damage to cells results in development of cancer and hereditary disease. The aim of the present study was to assess the effects of chronic low-dose X-ray radiation exposure on somatic DNA damage of interventional cardiologists working in high volume cardiac catheterization laboratories. For this analysis, we used peripheral lymphocytes and the assay for micronuclei (MNs), which is considered to be a reliable biological dosimeter for radiation exposure. We obtained peripheral blood from 62 physicians (mean age+/-se = 40.6+/-1.5 years): 31 interventional cardiologists (group I, exposed) and 31 age- and sex-matched clinical cardiologists (group II, nonexposed). Interventional cardiologists showed higher MN values (group I=20.5+/-1.6 vs. group II=12.8+/-1.3, P=0.001), although some overlap was apparent in the individual subject analysis. A correlation between years of professional activity and MN frequency value was detectable for interventional cardiologists (r=0.428, P=0.02) but not for clinical cardiologists (r=0.253, P=0.17). The results indicated that, overall, interventional cardiologists working in a high-volume catheterization laboratory have higher levels of somatic DNA damage when compared with clinical cardiologists working outside the catheterization laboratory. The amount of this damage varies and is only weakly related to the duration of professional exposure, which suggests that a dominant modulation of the underlying genetic substrate by environmental factors has a role in determining the harm in individual physicians. PMID- 15802492 TI - Protein breakdown in muscle from burned rats is blocked by insulin-like growth factor i and glycogen synthase kinase-3beta inhibitors. AB - We reported previously that IGF-I inhibits burn-induced muscle proteolysis. Recent studies suggest that activation of the phosphotidylinositol 3-kinase (PI3K)/Akt signaling pathway with downstream phosphorylation of Forkhead box O transcription factors is an important mechanism of IGF-I-induced anabolic effects in skeletal muscle. The potential roles of other mechanisms in the anabolic effects of IGF-I are less well understood. In this study we tested the roles of mammalian target of rapamycin and glycogen synthase kinase-3beta (GSK-3beta) phosphorylation as well as MAPK- and calcineurin-dependent signaling pathways in the anticatabolic effects of IGF-I by incubating extensor digitorum longus muscles from burned rats in the presence of IGF-I and specific signaling pathway inhibitors. Surprisingly, the PI3K inhibitors LY294002 and wortmannin reduced basal protein breakdown. No additional inhibition by IGF-I was noticed in the presence of LY294002 or wortmannin. Inhibition of proteolysis by IGF-I was associated with phosphorylation (inactivation) of GSK-3beta. In addition, the GSK 3beta inhibitors, lithium chloride and thiadiazolidinone-8, reduced protein breakdown in a similar fashion as IGF-I. Lithium chloride, but not thiadiazolidinone-8, increased the levels of phosphorylated Foxo 1 in incubated muscles from burned rats. Inhibitors of mammalian target of rapamycin, MAPK, and calcineurin did not prevent the IGF-I-induced inhibition of muscle proteolysis. Our results suggest that IGF-I inhibits protein breakdown at least in part through a PI3K/Akt/GSK3beta-dependent mechanism. Additional experiments showed that similar mechanisms were responsible for the effect of IGF-I in muscle from nonburned rats. Taken together with recent reports in the literature, the present results suggest that IGF-I inhibits protein breakdown in skeletal muscle by multiple mechanisms, including PI3K/Akt-mediated inactivation of GSK-3beta and Foxo transcription factors. PMID- 15802493 TI - Athyroid Pax8-/- mice cannot be rescued by the inactivation of thyroid hormone receptor alpha1. AB - The Pax8(-/-) mouse provides an ideal animal model to study the consequences of congenital hypothyroidism, because its only known defect is the absence of thyroid follicular cells. Pax8(-/-) mice are, therefore, completely athyroid in postnatal life and die around weaning unless they are substituted with thyroid hormones. As reported recently, Pax8(-/-) mice can also be rescued and survive to adulthood by the additional elimination of the entire thyroid hormone receptor alpha (TRalpha) gene, yielding Pax8(-/-)TRalpha(o/o) double-knockout animals. This observation has led to the hypothesis that unliganded TRalpha1 might be responsible for the lethal phenotype observed in Pax8(-/-) animals. In this study we report the generation of Pax8(-/-)TRalpha1(-/-) double-knockout mice that still express the non-T(3)-binding TR isoforms alpha2 and Deltaalpha2. These animals closely resemble the phenotype of Pax8(-/-) mice, including growth retardation and a completely distorted appearance of the pituitary with thyrotroph hyperplasia and hypertrophy, extremely high TSH mRNA levels, reduced GH mRNA expression, and the almost complete absence of lactotrophs. Like Pax8(-/ ) mice, Pax8(-/-)TRalpha1(-/-) compound mutants die around weaning unless they are substituted with thyroid hormones. These findings do not support the previous interpretation that the short life span of Pax8(-/-) mice is due to the negative effects of the TRalpha1 aporeceptor, but, rather, suggest a more complex mechanism involving TRalpha2 and an unliganded TR isoform. PMID- 15802495 TI - Evidence for ototopical glucocorticoid-induced decrease in hypothalamic-pituitary adrenal axis response and liver function. AB - To clarify whether ototopical glucocorticoid treatment is associated with impaired hypothalamic-pituitary-adrenal axis (HPA) activity and altered hepatic metabolism, one commercially available dexamethasone-containing ointment was tested. At present, very little is known about the effects of ototopical glucocorticoid treatment on HPA and liver function. Ten beagle dogs received two daily therapeutic doses of dexamethasone (0.6 mg/ear) in the outer auditory canal for 21 d in a single-blind, placebo-controlled study. Resting cortisol concentrations were assessed before, during, and after treatment using an RIA system. Adrenal function and HPA feedback sensitivity were measured by a standard dose (250 microg) ACTH stimulation test. Serum biochemical and hematological parameters were measured, whether ototopical glucocorticoids affect hepatic function was studied, and blood cell counts were made. Ototopical dexamethasone treatment induced a marked suppression (to about 100%) of resting plasma cortisol concentrations below the placebo effect (P < 0.0001) within the first 11 d, and these remained reduced during the entire treatment period up to d 19. As well, an ACTH stimulation test found a markedly reduced rise in plasma cortisol concentrations (P = 0.0004). Concomitantly, significant increases in serum activities of alkaline phosphatase, gamma-glutamyl transferase, alanine transaminase, and aspartate transaminase were detected. Moreover, we found a significant reduction in differential leukocyte counts of eosinophils and lymphocytes, whereas neutrophils increased. Although cortisol levels and hematological parameters returned to baseline 7 d after treatment cessation, liver enzyme activities remained elevated. In conclusion, these findings suggest that after ototopical application, dexamethasone is sufficiently absorbed from the auditory canal to suppress HPA function as well as to alter metabolic and hemopoietic profiles. Thus, in long-term treatment of otitis externa or media, the systemic adverse suppression of HPA has to be considered in relation to stress exposure, whereas changes in serum enzyme activities may not be interpreted as hepathopathy. PMID- 15802494 TI - Integrin alphaVbeta3 contains a cell surface receptor site for thyroid hormone that is linked to activation of mitogen-activated protein kinase and induction of angiogenesis. AB - Integrin alpha(V)beta(3) is a heterodimeric plasma membrane protein whose several extracellular matrix protein ligands contain an RGD recognition sequence. This study identifies integrin alpha(V)beta(3) as a cell surface receptor for thyroid hormone [L-T(4) (T(4))] and as the initiation site for T(4)-induced activation of intracellular signaling cascades. Integrin alpha(V)beta(3) dissociably binds radiolabeled T(4) with high affinity, and this binding is displaced by tetraiodothyroacetic acid, alpha(V)beta(3) antibodies, and an integrin RGD recognition site peptide. CV-1 cells lack nuclear thyroid hormone receptor, but express plasma membrane alpha(V)beta(3); treatment of these cells with physiological concentrations of T(4) activates the MAPK pathway, an effect inhibited by tetraiodothyroacetic acid, RGD peptide, and alpha(V)beta(3) antibodies. Inhibitors of T(4) binding to the integrin also block the MAPK mediated proangiogenic action of T(4). T(4)-induced phosphorylation of MAPK is inhibited by small interfering RNA knockdown of alpha(V) and beta(3). These findings suggest that T(4) binds to alpha(V)beta(3) near the RGD recognition site and show that hormone-binding to alpha(V)beta(3) has physiological consequences. PMID- 15802496 TI - Structure-function analysis of squirrel monkey FK506-binding protein 51, a potent inhibitor of glucocorticoid receptor activity. AB - FK506-binding protein 51 (FKBP51) and FKBP52 are large molecular weight immunophilins that are part of the mature glucocorticoid receptor (GR) heterocomplex. These proteins possess peptidyl-prolyl isomerase (PPIase) and tetratricopeptide repeats (TPR) domains that are important for modulation of GR activity. A naturally occurring animal model of glucocorticoid resistance, the squirrel monkey, results from the relative overexpression of FKBP51 that renders the GR in a low-affinity state. In vitro studies demonstrated that the squirrel monkey form of FKBP51 is greater than 6-fold more potent than human FKBP51 in this respect. The goals of these studies were to determine the roles of the TPR and PPIase domains in the inhibitory activity of squirrel monkey FKBP51 and to gain insight into structural features of squirrel monkey FKBP51 responsible for potent inhibition of dexamethasone-stimulated GR activity. Mutations in the TPR of squirrel monkey FKBP51 that inhibit association with heat shock protein 90 blocked GR inhibitory activity. Mutations that abrogate the PPIase activity of squirrel monkey FKBP51 had no effect on GR inhibitory activity. Chimeras of squirrel monkey and human FKBP51 were tested to identify domains responsible for their different inhibitory potencies. Amino acid differences in domains FK1 and FK2 between squirrel monkey and human FKBP51 contribute equally to the enhanced inhibitory activity of squirrel monkey FKBP51. Furthermore, squirrel monkey FKBP51 in which either FK1 or FK2 was deleted lacked GR inhibitory activity. Thus, the potent inhibitory activity of squirrel monkey FKBP51 involves both FK domains and the heat shock protein 90-binding TPR domain. PMID- 15802497 TI - Adrenocorticotropin evokes transient elevations in intracellular free calcium ([Ca2+]i) and increases basal [Ca2+]i in resting chondrocytes through a phospholipase C-dependent mechanism. AB - Both clinical and in vitro evidence points to the involvement of the melanocortin peptide, ACTH, in the terminal differentiation of chondrocytes. Terminal differentiation along the endochondral pathway is responsible for linear growth, but also plays a role in osteoarthritic cartilage degeneration. Chondrocyte terminal differentiation is associated with an incremental increase in chondrocyte basal intracellular free calcium ([Ca(2+)](i)), and ACTH agonism of melanocortin receptors is known to mobilize [Ca(2+)](i.) Using differentiated resting chondrocytes highly expressing type II collagen and aggrecan, we examined the influence of both ACTH and dexamethasone treatment on matrix gene transcription and [Ca(2+)](i). Resting chondrocytes treated concurrently with dexamethasone and ACTH expressed matrix gene transcripts in a pattern consistent with that of rapid terminal differentiation. Using the fluorescent Ca(2+) indicator, fura-2, we determined that ACTH evokes transient increases in [Ca(2+)](i) and elevates basal Ca(2+) levels in resting chondrocytes. The transient increases were initiated intracellularly, were abrogated by the phospholipase C-specific inhibitor, U73122, and were partly attenuated by myo inositol 1,4,5-triphosphate receptor inhibition via 10 mm caffeine. The initial intracellular release also resulted in store-operated calcium entry, presumably through store-operated channels. Dexamethasone priming increased both the initial ACTH-evoked [Ca(2+)](i) release and the subsequent store-operated calcium entry. These data demonstrate roles for ACTH and glucocorticoid in the regulation of chondrocyte terminal differentiation. Because the actions of ACTH are mediated through known G protein-coupled receptors, the melanocortin receptors, these data may provide a new therapeutic target in the treatment of growth deficiencies and cartilage degeneration. PMID- 15802498 TI - Palmitate-induced interleukin 6 production is mediated by protein kinase C and nuclear-factor kappaB activation and leads to glucose transporter 4 down regulation in skeletal muscle cells. AB - The mechanisms by which elevated levels of free fatty acids cause insulin resistance are not well understood. In addition, accumulating evidence suggests a link between inflammation and type 2 diabetes. Here, we report that exposure of C2C12 skeletal muscle cells to 0.5 mm palmitate results in increased mRNA levels (3.5-fold induction; P < 0.05) and secretion (control 375 +/- 57 vs. palmitate 1129 +/- 177 pg/ml; P < 0.001) of the proinflammatory cytokine IL-6. Palmitate increased nuclear factor-kappaB activation and coincubation of the cells with palmitate and the nuclear factor-kappaB inhibitor pyrrolidine dithiocarbamate prevented both IL-6 expression and secretion. Furthermore, incubation of palmitate-treated cells with calphostin C, a strong and specific inhibitor of protein kinase C, and phorbol myristate acetate, that down-regulates protein kinase C in long-term incubations, abolished induction of IL-6 production. Finally, exposure of skeletal muscle cells to palmitate caused a fall in the mRNA levels of glucose transporter 4 and insulin-stimulated glucose uptake, whereas in the presence of anti-IL-6 antibody, which neutralizes the biological activity of mouse IL-6 in cell culture, these reductions were prevented. These findings suggest that IL-6 may mediate several of the prodiabetic effects of palmitate. PMID- 15802499 TI - Differential adaptive responses to chronic stress of maternally stressed male mice offspring. AB - It is well established that stress in early life can alter the activity of the hypothalamus-pituitary-adrenal (HPA) axis, but most studies to date have focused on HPA reactivity in response to a single acute stress. The present study addressed whether stress in pregnant mice could influence the adaptive responses of their offspring to chronic stress. Male offspring were exclusively used in this study. Elevated plus maze tests revealed that 14 d of repeated restraint stress (6 h per day; from postnatal d 50-63) significantly increased anxiety-like behavior in maternally stressed mice. NBI 27914, a CRH receptor antagonist, completely eliminated anxiety-related behaviors in a dose-dependent manner, indicating an involvement of a hyperactive CRH system. In accordance with increased anxiety, CRH contents in the hypothalamus and amygdala were significantly higher in these mice. Despite an increased basal activity of the CRH-ACTH system, the combination of chronic prenatal and postnatal stress resulted in a significant reduction of basal plasma corticosterone level, presumably because of a defect in adrenal function. Along with alterations in hypothalamic and hippocampal corticosteroid receptors, it was also demonstrated that a dysfunction in negative feedback inhibition of the HPA axis could be deteriorated by chronic stress in maternally stressed male mice. Taken together, these results indicate that exposure to maternal stress in the womb can affect an animal's coping capacity to chronic postnatal stress. PMID- 15802500 TI - Fetal programming: prenatal testosterone treatment causes intrauterine growth retardation, reduces ovarian reserve and increases ovarian follicular recruitment. AB - Exposure to testosterone (T) during d 30-90 of fetal life results in low-birth weight offspring, hypergonadotropism, multifollicular ovaries, and early cessation of cyclicity. The multifollicular phenotype may result from failure of follicles to regress and consequent follicular persistence or, alternatively, increased follicular recruitment. We tested the hypothesis that prenatal exposure to excess T causes intrauterine growth retardation and increases ovarian follicular recruitment. Time-mated pregnant ewes were treated with 100 mg T propionate in cottonseed oil or vehicle twice weekly from d 30-90 of gestation. Ewes were euthanized near term, from d 139-141 of gestation (term is 147 d). After determining fetal measures and organ weights, ovaries were removed from fetuses of control and T-treated dams, and follicular distribution in each ovary was determined by morphometric quantification. Total number and percentage distribution of the various classes of follicles (primordial, primary, preantral, and antral follicles) were compared between treatment groups. Prenatally T treated female fetuses were smaller in size, had an increased head circumference to fetal weight ratio (P < 0.01), increased adrenal to fetal weight ratio (P < 0.05), decreased number of follicles (P < 0.05), a decrease in percentage of primordial follicles (P < 0.001), and a corresponding increase in the remaining classes of follicles (P < 0.05). Ovarian findings support decreased ovarian reserve and enhanced follicular recruitment, potential contributors of early reproductive failure. The extent to which metabolic changes associated with intrauterine growth retardation contribute toward altered trajectory of ovarian folliculogenesis remains to be determined. PMID- 15802501 TI - Enhancement of tumor necrosis factor-alpha-induced growth inhibition by insulin like growth factor-binding protein-5 (IGFBP-5), but not IGFBP-3 in human breast cancer cells. AB - Expression of IGF-binding protein-3 (IGFBP-3) and IGFBP-5 in human breast cancer cells induces apoptosis and is associated with modulations in Bcl-2 proteins, suggesting that these IGFBPs induce an intrinsic apoptotic pathway. In this study we demonstrate that although both IGFBPs induced the activation of caspase-8 and caspase-9, the expression of IGFBP-5, but not IGFBP-3, sensitized MDA-MB-231 breast cancer cells to the inhibitory effects of TNFalpha. This sensitivity to TNFalpha was associated with a block in nuclear factor-kappaB-mediated cell survival signals. IGFBP-5 expression was also associated with a caspase-8 independent activation of Bid, increased levels of cytosolic second mitochondria derived activator of caspase (Smac)/direct inhibitor of apoptosis proteins (IAP) binding protein with low pI (DIABLO), and an enhanced phosphorylation of c-Jun N terminal kinase, both basally and in response to TNFalpha. These results suggest that IGFBP-5 expression may influence extrinsic apoptotic pathways via a differential modulation of downstream cell survival and cell death pathways. Furthermore, although IGFBP-3 and IGFBP-5 share much structural and functional homology, they can modulate distinct apoptotic pathways in human breast cancer cells. PMID- 15802502 TI - Growth hormone modulates thymocyte development in vivo through a combined action of laminin and CXC chemokine ligand 12. AB - Previous evidence indicates that GH modulates thymic cell migration. In this study we approached this issue in vivo, studying thymocyte migration in GH transgenic animals and in normal mice treated intrathymically with GH. Extracellular matrix and chemokines are involved in thymocyte migration. In this respect, thymocyte adhesion to laminin was higher in GH-treated animals than controls, and the numbers of migrating cells in laminin-coated Transwells was higher in GH-transgenic and GH-injected mice. Additionally, CXC chemokine ligand 12 (CXCL12)-driven migration was higher in GH-Tg and GH-treated animals compared with controls. Interestingly, although CXCR4 expression on thymocytes did not change in GH-Tg mice, the CXCL12 intrathymic contents were higher. We found that CXCL12, in conjunction with laminin, would additionally enhance the migration of thymocytes previously exposed to high concentrations of GH in vivo. Lastly, there was an augmentation of recent thymic emigrants in lymph nodes from GH-Tg and GH injected animals. In conclusion, enhanced thymocyte migration in GH transgenic mice as well as GH-injected mice results at least partially from a combined action of laminin and CXCL12. Considering that GH is presently being used as an adjuvant therapeutic agent in immunodeficiencies, including AIDS, the concepts defined herein provide important background knowledge for future GH-based immune interventions. PMID- 15802503 TI - Origins of host-specific populations of the blast pathogen Magnaporthe oryzae in crop domestication with subsequent expansion of pandemic clones on rice and weeds of rice. AB - Rice, as a widely and intensively cultivated crop, should be a target for parasite host shifts and a source for shifts to co-occurring weeds. Magnaporthe oryzae, of the M. grisea species complex, is the most important fungal pathogen of rice, with a high degree of host specificity. On the basis of 10 loci from six of its seven linkage groups, 37 multilocus haplotypes among 497 isolates of M. oryzae from rice and other grasses were identified. Phylogenetic relationships among isolates from rice (Oryza sativa), millet (Setaria spp.), cutgrass (Leersia hexandra), and torpedo grass (Panicum repens) were predominantly tree like, consistent with a lack of recombination, but from other hosts were reticulate, consistent with recombination. The single origin of rice-infecting M. oryzae followed a host shift from a Setaria millet and was closely followed by additional shifts to weeds of rice, cutgrass, and torpedo grass. Two independent estimators of divergence time indicate that these host shifts predate the Green Revolution and could be associated with rice domestication. The rice-infecting lineage is characterized by high copy number of the transposable element MGR586 (Pot3) and, except in two haplotypes, by a loss of AVR-Co39. Both mating types have been retained in ancestral, well-distributed rice-infecting haplotypes 10 (mainly temperate) and 14 (mainly tropical), but only one mating type was recovered from several derived, geographically restricted haplotypes. There is evidence of a common origin of both ACE1 virulence genotypes in haplotype 14. Host-haplotype association is evidenced by low pathogenicity on hosts associated with other haplotypes. PMID- 15802504 TI - Spontaneous mutations in the ammonium transport gene AMT4 of Chlamydomonas reinhardtii. AB - Evidence in several microorganisms indicates that Amt proteins are gas channels for NH(3) and CH(3)NH(2), and this has been confirmed structurally. Chlamydomonas reinhardtii has at least four AMT genes, the most reported for a microorganism. Under nitrogen-limiting conditions all AMT genes are transcribed and Chlamydomonas is sensitive to methylammonium toxicity. All 16 spontaneous methylammonium-resistant mutants that we analyzed had defects in accumulation of [(14)C]methylammonium. Genetic crosses indicated that 12 had lesions in a single locus, whereas two each had lesions in other loci. Lesions in different loci were correlated with different degrees of defect in [(14)C]methylammonium uptake. One mutant in the largest class had an insert in the AMT4 gene, and the insert cosegregated with methylammonium resistance in genetic crosses. The other 11 strains in this class also had amt4 lesions, which we characterized at the molecular level. Properties of the amt4 mutants were clearly different from those of rh1 RNAi lines. They indicated that the physiological substrates for Amt and Rh proteins, the only two members of their protein superfamily, are NH(3) and CO(2), respectively. PMID- 15802505 TI - DNA variation at the invertase locus invGE/GF is associated with tuber quality traits in populations of potato breeding clones. AB - Starch and sugar content of potato tubers are quantitative traits, which are models for the candidate gene approach for identifying the molecular basis of quantitative trait loci (QTL) in noninbred plants. Starch and sugar content are also important for the quality of processed products such as potato chips and French fries. A high content of the reducing sugars glucose and fructose results in inferior chip quality. Tuber starch content affects nutritional quality. Functional and genetic models suggest that genes encoding invertases control, among other things, tuber sugar content. The invGE/GF locus on potato chromosome IX consists of duplicated invertase genes invGE and invGF and colocalizes with cold-sweetening QTL Sug9. DNA variation at invGE/GF was analyzed in 188 tetraploid potato cultivars, which have been assessed for chip quality and tuber starch content. Two closely correlated invertase alleles, invGE-f and invGF-d, were associated with better chip quality in three breeding populations. Allele invGF-b was associated with lower tuber starch content. The potato invertase gene invGE is orthologous to the tomato invertase gene Lin5, which is causal for the fruit-sugar-yield QTL Brix9-2-5, suggesting that natural variation of sugar yield in tomato fruits and sugar content of potato tubers is controlled by functional variants of orthologous invertase genes. PMID- 15802506 TI - The phosphatase subunit tap42 functions independently of target of rapamycin to regulate cell division and survival in Drosophila. AB - The protein phosphatase 2A (PP2A) regulatory subunit Tap42 is essential for target of rapamycin (TOR)-mediated signaling in yeast, but its role in higher eukaryotes has not been established. Here we show that Tap42 does not contribute significantly to TOR signaling in Drosophila, as disruption of the Tap42 gene does not cause defects in cell growth, metabolism, or S6-kinase activity characteristic of TOR inactivation. In addition, Tap42 is not required for increased cell growth in response to activation of TOR signaling. Instead, we find that Tap42 mutations cause disorganization of spindle microtubules in larval neuroblasts, leading to a preanaphase mitotic arrest in these cells. Loss of Tap42 ultimately results in increased JNK signaling, caspase activation, and cell death. These phenotypes are associated with increased accumulation and nuclear localization of PP2A in Tap42 mutant cells. Our results demonstrate that the role of Tap42 in TOR signaling has not been conserved in higher eukaryotes, indicating fundamental differences in the mechanisms of TOR signaling between yeast and higher eukaryotes. PMID- 15802507 TI - Subdivision of large introns in Drosophila by recursive splicing at nonexonic elements. AB - Many genes with important roles in development and disease contain exceptionally long introns, but special mechanisms for their expression have not been investigated. We present bioinformatic, phylogenetic, and experimental evidence in Drosophila for a mechanism that subdivides many large introns by recursive splicing at nonexonic elements and alternative exons. Recursive splice sites predicted with highly stringent criteria are found at much higher frequency than expected in the sense strands of introns >20 kb, but they are found only at the expected frequency on the antisense strands, and they are underrepresented within introns <10 kb. The predicted sites in long introns are highly conserved between Drosophila melanogaster and Drosophila pseudoobscura, despite extensive divergence of other sequences within the same introns. These patterns of enrichment and conservation indicate that recursive splice sites are advantageous in the context of long introns. Experimental analyses of in vivo processing intermediates and lariat products from four large introns in the unrelated genes kuzbanian, outspread, and Ultrabithorax confirmed that these introns are removed by a series of recursive splicing steps using the predicted nonexonic sites. Mutation of nonexonic site RP3 within Ultrabithorax also confirmed that recursive splicing is the predominant processing pathway even with a shortened version of the intron. We discuss currently known and potential roles for recursive splicing. PMID- 15802508 TI - Chromosome sorting in tetraploid wheat and its potential for genome analysis. AB - This study evaluates the potential of flow cytometry for chromosome sorting in durum wheat (Triticum turgidum Desf. var. durum, 2n = 4x = 28). Histograms of fluorescence intensity (flow karyotypes) obtained after the analysis of DAPI stained chromosomes consisted of three peaks. Of these, one represented chromosome 3B, a small peak corresponded to chromosomes 1A and 6A, and a large peak represented the remaining 11 chromosomes. Chromosomes sorted onto microscope slides were identified after fluorescence in situ hybridization (FISH) with probes for GAA microsatellite, pSc119.2, and Afa repeats. Genomic distribution of these sequences was determined for the first time in durum wheat and a molecular karyotype has been developed for this crop. Flow karyotyping in double ditelosomic lines of durum wheat revealed that the lines facilitated sorting of any arm of the wheat A- and B-genome chromosomes. Compared to hexaploid wheat, flow karyotype of durum wheat is less complex. This property results in better discrimination of telosomes and high purities in sorted fractions, ranging from 90 to 98%. We have demonstrated that large insert libraries can be created from DNA purified using flow cytometry. This study considerably expands the potential of flow cytogenetics for use in wheat genomics and opens the possibility of sequencing the genome of this important crop one chromosome arm at a time. PMID- 15802509 TI - The program of sex chromosome pairing in meiosis is highly conserved across marsupial species: implications for sex chromosome evolution. AB - Marsupials present a series of genetic and chromosomal features that are highly conserved in very distant species. One of these features is the absence of a homologous region between X and Y chromosomes. According to this genetic differentiation, sex chromosomes do not synapse during the first meiotic prophase in males, and a special structure, the dense plate, maintains sex chromosome association. In this report we present results on the process of meiotic sex chromosome pairing obtained from three different species, Thylamys elegans, Dromiciops gliroides, and Rhyncholestes raphanurus, representing the three orders of American marsupials. We have investigated the relationships between the axial structures organized along sex chromosomes and the formation of the dense plate. We found that in the three species the dense plate arises as a modification of sex chromosomal axial elements, but without the involvement of other meiotic axial structures, such as the cohesin axes. Considering the phylogenetic relationships among the marsupials studied here, our data reinforce the idea that the dense plate emerged early in marsupial evolution as an efficient mechanism to ensure the association of the nonhomologous sex chromosomes. This situation could have influenced the further evolution of sex chromosomes in marsupials. PMID- 15802510 TI - A genomics approach to the detection of positive selection in cattle: adaptive evolution of the T-cell and natural killer cell-surface protein CD2. AB - The detection of adaptive evolution at the molecular level is of interest not only as an insight into the process of evolution but also because of its functional implications for genes of interest. Here, we present the first genomics approach to detecting positive selection operating on the Bos taurus lineage, an important domestic species. This analysis led to the identification of the T-cell and natural killer (NK) cell receptor cluster of differentiation 2 (CD2) as having a strong signal of selection. Further detailed investigation of CD2 revealed that this gene was subject to positive selection during the evolution of a number of mammalian lineages. Moreover, we show that selection has operated primarily on the extracellular domain of CD2 and discuss the implications of this for an important regulator of the adaptive immune response. PMID- 15802511 TI - A high-frequency null mutant of an odorant-binding protein gene, Obp57e, in Drosophila melanogaster. AB - We have found a null mutant of an odorant-binding protein, Obp57e, in Drosophila melanogaster. This frameshift mutation, which is a 10-bp deletion in the coding region, is at a high frequency in the Kyoto population and is also present in Taiwan and Africa. We have sequenced a 1.5-kb region including the tandemly duplicated gene, Obp57d, from 16 inbred lines sampled in Kyoto, Japan. The analyses showed a peak of nucleotide diversity and strong linkage disequilibrium around this mutation. This pattern suggests an elevated mutation rate or an influence of balancing selection in this region. The level of nucleotide divergence between D. melanogaster and D. simulans does not support the former possibility. Thus, this presence/absence polymorphism may be due to balancing selection, which takes advantage of the relatively weak functional constraint in members of a large gene family. In addition, the Obp57d gene region showed an excess of high-frequency-derived mutants that is consistent with a pattern predicted under positive natural selection. PMID- 15802512 TI - Selection theory for marker-assisted backcrossing. AB - Marker-assisted backcrossing is routinely applied in breeding programs for gene introgression. While selection theory is the most important tool for the design of breeding programs for improvement of quantitative characters, no general selection theory is available for marker-assisted backcrossing. In this treatise, we develop a theory for marker-assisted selection for the proportion of the genome originating from the recurrent parent in a backcross program, carried out after preselection for the target gene(s). Our objectives were to (i) predict response to selection and (ii) give criteria for selecting the most promising backcross individuals for further backcrossing or selfing. Prediction of response to selection is based on the marker linkage map and the marker genotype of the parent(s) of the backcross population. In comparison to standard normal distribution selection theory, the main advantage of our approach is that it considers the reduction of the variance in the donor genome proportion due to selection. The developed selection criteria take into account the marker genotype of the candidates and consider whether these will be used for selfing or backcrossing. Prediction of response to selection is illustrated for model genomes of maize and sugar beet. Selection of promising individuals is illustrated with experimental data from sugar beet. The presented approach can assist geneticists and breeders in the efficient design of gene introgression programs. PMID- 15802513 TI - The C-terminal half of Saccharomyces cerevisiae Mad1p mediates spindle checkpoint function, chromosome transmission fidelity and CEN association. AB - The evolutionarily conserved spindle checkpoint is a key mechanism ensuring high fidelity chromosome transmission. The checkpoint monitors attachment between kinetochores and mitotic spindles and the tension between sister kinetochores. In the absence of proper attachment or tension, the spindle checkpoint mediates cell cycle arrest prior to anaphase. Saccharomyces cerevisiae Mad1p is required for the spindle checkpoint and for chromosome transmission fidelity. Moreover, Mad1p associates with the nuclear pore complex (NPC) and is enriched at kinetochores upon checkpoint activation. Using partial mad1 deletion alleles we determined that the C-terminal half of Mad1p is necessary and sufficient for checkpoint activation in response to microtubule depolymerizing agents, high-fidelity transmission of a reporter chromosome fragment, and in vivo association with centromeres, but not for robust NPC association. Thus, spindle checkpoint activation and chromosome transmission fidelity correlate and these Mad1p functions likely involve kinetochore association but not robust NPC association. These studies are the basis for elucidating the role of protein complexes containing Mad1p in the spindle checkpoint pathway and in maintaining genome stability in S. cerevisiae and other systems. PMID- 15802514 TI - Mutations in snail family genes enhance craniosynostosis of Twist1 haplo insufficient mice: implications for Saethre-Chotzen Syndrome. AB - In Drosophila, mutations in the Twist gene interact with mutations in the Snail gene. We show that the mouse Twist1 mutation interacts with Snai1 and Snai2 mutations to enhance aberrant cranial suture fusion, demonstrating that genetic interactions between genes of the Twist and Snail families have been conserved during evolution. PMID- 15802515 TI - On the mutation rate of herpes simplex virus type 1. AB - All seven DNA-based microbes for which carefully established mutation rates and mutational spectra were previously available displayed a genomic mutation rate in the neighborhood of 0.003 per chromosome replication. The pathogenic mammalian DNA virus herpes simplex type 1 has an estimated genomic mutation rate compatible with that value. PMID- 15802516 TI - The Bombyx mori karyotype and the assignment of linkage groups. AB - Lepidopteran species have a relatively high number of small holocentric chromosomes (Bombyx mori, 2n = 56). Chromosome identification has long been hampered in this group by the high number and by the absence of suitable markers like centromere position and chromosome bands. In this study, we carried out fluorescence in situ hybridization (FISH) on meiotic chromosome complements using genetically mapped B. mori bacterial artificial chromosomes (BACs) as probes. The combination of two to four either green or red fluorescence-labeled probes per chromosome allowed us to recognize unequivocally each of the 28 bivalents of the B. mori karyotype by its labeling pattern. Each chromosome was assigned one of the already established genetic linkage groups and the correct orientation in the chromosome was defined. This facilitates physical mapping of any other sequence and bears relevance for the ongoing B. mori genome projects. Two-color BAC-FISH karyotyping overcomes the problem of chromosome recognition in organisms where conventional banding techniques are not available. PMID- 15802517 TI - Sensitivity to phosphonoacetic acid: a new phenotype to probe DNA polymerase delta in Saccharomyces cerevisiae. AB - A mutant allele (pol3-L612M) of the DNA polymerase delta gene in Saccharomyces cerevisiae that confers sensitivity to the antiviral drug phosphonoacetic acid (PAA) was constructed. We report that PAA-sensitivity tagging DNA polymerases is a useful method for selectively and reversibly inhibiting one type of DNA polymerase. Our initial studies reveal that replication by the L612M-DNA pol delta requires Rad27 flap endonuclease activity since the pol3-L612M strain is not viable in the absence of RAD27 function. The L612M-DNA pol delta also strongly depends on mismatch repair (MMR). Reduced viability is observed in the absence of any of the core MMR proteins-Msh2, Mlh1, or Pms1-and severe sensitivity to PAA is observed in the absence of the core proteins Msh6 or Exo1, but not Msh3. We propose that pol3-L612M cells need the Rad27 flap endonuclease and MMR complexes composed of Msh2/Msh6, Mlh1/Pms1, and Exo1 for correct processing of Okazaki fragments. PMID- 15802518 TI - Dynein promotes achiasmate segregation in Schizosaccharomyces pombe. AB - Most organisms use crossovers (chiasmata) to maintain physical connections between homologous chromosomes that ensure their proper segregation at the first meiotic division. The fission yeast Schizosaccharomyces pombe has a residual ability to segregate homologous chromosomes in the absence of meiotic recombination (achiasmate segregation). Using cytologically tagged chromosomes, we established a role for the microtubule motor dynein in meiotic chromosome segregation. Dhc1, the motor subunit of dynein, is required for chromosome segregation in both the presence and the absence of recombination. Dlc1, a member of the Tctex-1 dynein light-chain family, preferentially affects the segregation of achiasmate chromosomes. Dlc1 is the first identified protein, outside of Drosophila, that preferentially affects achiasmate chromosome segregation. We discuss possible roles of the dynein motor in this process. PMID- 15802519 TI - Interaction of the Saccharomyces cerevisiae cortical actin patch protein Rvs167p with proteins involved in ER to Golgi vesicle trafficking. AB - We have used affinity chromatography to identify two proteins that bind to the SH3 domain of the actin cytoskeleton protein Rvs167p: Gyp5p and Gyl1p. Gyp5p has been shown to be a GTPase activating protein (GAP) for Ypt1p, a Rab GTPase involved in ER to Golgi trafficking; Gyl1p is a protein that resembles Gyp5p and has recently been shown to colocalize with and belong to the same protein complex as Gyp5p. We show that Gyl1p and Gyp5p interact directly with each other, likely through their carboxy-terminal coiled-coil regions. In assays of GAP activity, Gyp5p had GAP activity toward Ypt1p and we found that this activity was stimulated by the addition of Gyl1p. Gyl1p had no GAP activity toward Ypt1p. Genetic experiments suggest a role for Gyp5p and Gyl1p in ER to Golgi trafficking, consistent with their biochemical role. Since Rvs167p has a previously characterized role in endocytosis and we have shown here that it interacts with proteins involved in Golgi vesicle trafficking, we suggest that Rvs167p may have a general role in vesicle trafficking. PMID- 15802520 TI - Crossover interference on nucleolus organizing region-bearing chromosomes in Arabidopsis. AB - In most eukaryotes, crossovers are not independently distributed along the length of a chromosome. Instead, they appear to avoid close proximity to one another--a phenomenon known as crossover interference. Previously, for three of the five Arabidopsis chromosomes, we measured the strength of interference and suggested a model wherein some crossovers experience interference while others do not. Here we show, using the same model, that the fraction of interference-insensitive crossovers is significantly smaller on the remaining two chromosomes. Since these two chromosomes bear the Arabidopsis NOR domains, the possibility that these chromosomal regions influence interference is discussed. PMID- 15802521 TI - Theoretical basis for the identification of allelic variants that encode drug efficacy and toxicity. AB - Almost all drugs that produce a favorable response (efficacy) may also produce adverse effects (toxicity). The relative strengths of drug efficacy and toxicity that vary in human populations are controlled by the combined influences of multiple genes and environmental influences. Genetic mapping has proven to be a powerful tool for detecting and identifying specific DNA sequence variants on the basis of the haplotype map (HapMap) constructed from single-nucleotide polymorphisms (SNPs). In this article, we present a novel statistical model for sequence mapping of two different but related drug responses. This model is incorporated by mathematical functions of drug response to varying doses or concentrations and the statistical device used to model the correlated structure of the residual (co)variance matrix. We implement a closed-form solution for the EM algorithm to estimate the population genetic parameters of SNPs and the simplex algorithm to estimate the curve parameters describing the pharmacodynamic changes of different genetic variants and matrix-structuring parameters. Extensive simulations are performed to investigate the statistical properties of our model. The implications of our model in pharmacogenetic and pharmacogenomic research are discussed. PMID- 15802522 TI - Using Drosophila to decipher how mutations associated with human branchio-oto renal syndrome and optical defects compromise the protein tyrosine phosphatase and transcriptional functions of eyes absent. AB - Eyes absent (EYA) proteins are defined by a conserved C-terminal EYA domain (ED) that both contributes to its function as a transcriptional coactivator by mediating protein-protein interactions and possesses intrinsic protein tyrosine phosphatase activity. Mutations in human EYA1 result in an autosomal dominant disorder called branchio-oto-renal (BOR) syndrome as well as congenital cataracts and ocular defects (OD). Both BOR- and OD-associated missense mutations alter residues in the conserved ED as do three missense mutations identified from Drosophila eya alleles. To investigate the molecular mechanisms whereby these mutations disrupt EYA function, we tested their activity in a series of assays that measured in vivo function, phosphatase activity, transcriptional capability, and protein-protein interactions. We find that the OD-associated mutations retain significant in vivo activity whereas those derived from BOR patients show a striking decrease or loss of in vivo functionality. Protein-protein interactions, either with its partner transcription factor Sine oculis or with EYA itself, were not significantly compromised. Finally, the results of the biochemical assays suggest that both loss of protein tyrosine phosphatase activity and reduced transcriptional capability contribute to the impaired EYA function associated with BOR/OD syndrome, thus shedding new light into the molecular mechanisms underlying this disease. PMID- 15802524 TI - A mitogen-activated protein kinase pathway essential for mating and contributing to vegetative growth in Neurospora crassa. AB - MAP kinases homologous to Saccharomyces cerevisiae Fus3p/Kss1p have been identified in plant pathogenic fungi and are required for pathogenicity and sexual reproduction. To better understand the role of MAP kinase signaling in Neurospora crassa, and to identify downstream target genes of the pathway, we isolated, cloned, and disrupted the FUS3 homolog mak-2. Ste12p is a transcription factor target of Fus3p that activates genes of the mating pathway in yeast, and we also characterized the N. crassa STE12 homolog pp-1. The mak-2 and pp-1 mutants have reduced growth rate, produce short aerial hyphae, and fail to develop protoperithecia. In addition, ascospores carrying null mutations of either gene are inviable. Subtractive cloning was used to isolate genes having reduced expression in the mak-2 mutant. Expression of some of these genes is protoperithecia specific and three of them are part of a gene cluster potentially involved in the production of a polyketide secondary metabolite. Microarray analysis was used to extend the analysis of gene expression in mak-2 and pp-1 mutants. The role of the MAP kinase pathway in both sexual and asexual development as well as secondary metabolism is consistent with the dual regulation of the mating process and pathogencity observed in fungal pathogens. PMID- 15802523 TI - A postsynaptic role for Rhp55/57 that is responsible for cell death in Deltarqh1 mutants following replication arrest in Schizosaccharomyces pombe. AB - Following replication arrest, multiple cellular responses are triggered to maintain genomic integrity. In fission yeast, the RecQ helicase, Rqh1, plays a critical role in this process. This is demonstrated in Deltarqh1 cells that, following treatment with hydroxyurea (HU), undergo an aberrant mitosis leading to cell death. Previous data suggest that Rqh1 functions with homologous recombination (HR) in recovery from replication arrest. We have found that loss of the HR genes rhp55(+) or rhp57(+), but not rhp51(+) or rhp54(+), suppresses the HU sensitivity of Deltarqh1 cells. Much of this suppression requires Rhp51 and Rhp54. In addition, this suppression is partially dependent on swi5(+). In budding yeast, overexpressing Rad51 (the Rhp51 homolog) minimized the need for Rad55/57 (Rhp55/57) in nucleoprotein filament formation. We overexpressed Rhp51 in Schizosaccharomyces pombe and found that it greatly reduced the requirement for Rhp55/57 in recovery from DNA damage. However, overexpressing Rhp51 did not change the Deltarhp55 suppression of the HU sensitivity of Deltarqh1, supporting an Rhp55/57 function during HR independent of nucleoprotein filament formation. These results are consistent with Rqh1 playing a role late in HR following replication arrest and provide evidence for a postsynaptic function for Rhp55/57. PMID- 15802525 TI - A general framework for statistical linkage analysis in multivalent tetraploids. AB - In multivalent polyploids, simultaneous pairings among homologous chromosomes at meiosis result in a unique cytological phenomenon-double reduction. Double reduction casts an impact on chromosome evolution in higher plants, but because of its confounded effect on the pattern of gene cosegregation, it complicates linkage analysis and map construction with polymorphic molecular markers. In this article, we have proposed a general statistical model for simultaneously estimating the frequencies of double reduction, the recombination fraction, and optimal parental linkage phases between any types of markers, both fully and partially informative, or dominant and codominant, for a tetraploid species that undergoes only multivalent pairing. This model provides an in-depth extension of our earlier linkage model that was built upon Fisher's classifications for different gamete formation modes during the polysomic inheritance of a multivalent polyploid. By implementing a two-stage hierarchical EM algorithm, we derived a closed-form solution for estimating the frequencies of double reduction through the estimation of gamete mode frequencies and the recombination fraction. We performed different settings of simulation studies to demonstrate the statistical properties of our model for estimating and testing double reduction and the linkage in multivalent tetraploids. As shown by a comparative analysis, our model provides a general framework that covers existing statistical approaches for linkage mapping in polyploids that are predominantly multivalent. The model will have great implications for understanding the genome structure and organization of polyploid species. PMID- 15802526 TI - Combined linkage and association mapping of quantitative trait loci by multiple markers. AB - Using multiple diallelic markers, variance component models are proposed for high resolution combined linkage and association mapping of quantitative trait loci (QTL) based on nuclear families. The objective is to build a model that may fully use marker information for fine association mapping of QTL in the presence of prior linkage. The measures of linkage disequilibrium and the genetic effects are incorporated in the mean coefficients and are decomposed into orthogonal additive and dominance effects. The linkage information is modeled in variance-covariance matrices. Hence, the proposed methods model both association and linkage in a unified model. On the basis of marker information, a multipoint interval mapping method is provided to estimate the proportion of allele sharing identical by descent (IBD) and the probability of sharing two alleles IBD at a putative QTL for a sib-pair. To test the association between the trait locus and the markers, both likelihood-ratio tests and F-tests can be constructed on the basis of the proposed models. In addition, analytical formulas of noncentrality parameter approximations of the F-test statistics are provided. Type I error rates of the proposed test statistics are calculated to show their robustness. After comparing with the association between-family and association within-family (AbAw) approach by Abecasis and Fulker et al., it is found that the method proposed in this article is more powerful and advantageous based on simulation study and power calculation. By power and sample size comparison, it is shown that models that use more markers may have higher power than models that use fewer markers. The multiple-marker analysis can be more advantageous and has higher power in fine mapping QTL. As an application, the Genetic Analysis Workshop 12 German asthma data are analyzed using the proposed methods. PMID- 15802528 TI - Identification of novel vascular endothelial-specific genes by the microarray analysis of the zebrafish cloche mutants. AB - The zebrafish cloche (clo) mutation affects the earliest known step in differentiation of blood and endothelial cells in vertebrates. We established clo/gata1-GFP transgenic line with erythroid-specific green fluorescent protein (GFP) expression, which allowed differentiation of clo and wild-type siblings at the midsomitogenesis stages before morphologically visible phenotypes appeared. To discover novel genes potentially involved in hematopoietic and vascular development, we performed microarray analysis of more than 15,000 zebrafish genes or expressed sequence tags (ESTs) in clo mutant embryos. We isolated the full length sequences and determined the expression patterns for 8 novel cDNAs that were significantly down-regulated in clo-/- embryos. Dual specificity phosphatase 5 (dusp5), cadherin 5 (cdh5; VE-cadherin), aquaporin 8 (aqp8), adrenomedullin receptor (admr), complement receptor C1qR-like (crl), scavenger receptor class F, member 1 (scarf1), and ETS1-like protein (etsrp) were specifically expressed in the vascular endothelial cells, while retinol binding protein 4 (rbp4) was expressed in the yolk syncytial layer and the hypochord. Further functional studies of these novel genes should help to elucidate critical early steps leading to the formation of vertebrate blood vessels. PMID- 15802527 TI - SDX-101, the R-enantiomer of etodolac, induces cytotoxicity, overcomes drug resistance, and enhances the activity of dexamethasone in multiple myeloma. AB - In this study we report that R-etodolac (SDX-101), at clinically relevant concentrations, induces potent cytotoxicity in drug-sensitive multiple myeloma (MM) cell lines, as well as in dexamethasone (MM.1R)-, doxorubicin (Dox40/RPMI8226)-, and bortezomib (DHL4)-resistant cell lines. Immunoblot analysis demonstrates that R-etodolac induces apoptosis characterized by caspase 8, -9, and -3 and PARP (poly-ADP [adenosine diphosphate]-ribose polymerase) cleavage and down-regulation of cyclin D1 expression. Subcytotoxic doses of R etodolac up-regulate myeloid cell leukemia-1 proapoptotic variant (Mcl-1S), while enhancing dexamethasone (Dex)-induced caspase activation and apoptosis. The combination of R-etodolac with Dex results in a highly synergistic cytotoxic effect. R-etodolac also induces apoptosis against primary cells isolated from patients with MM refractory to chemotherapy. Although interleukin 6 (IL-6) and insulin-like growth factor-1 (IGF-1) abrogate Dex-induced MM cell cytotoxicity, neither IL-6 nor IGF-1 protects against R-etodolac-induced cytotoxicity in MM cells. R-etodolac also inhibits viability of MM cells adherent to bone marrow stromal cells (BMSCs), thereby overcoming a mechanism of drug resistance commonly observed with other conventional chemotherapeutic agents. Our data, therefore, indicate that R-etodolac circumvents drug resistance in MM cells at clinically relevant concentrations, targets Mcl-1, and can be synergistically combined with Dex. PMID- 15802530 TI - Oncogenes as molecular targets in lymphoma. PMID- 15802529 TI - CXCL9 inhibits eosinophil responses by a CCR3- and Rac2-dependent mechanism. AB - Recently, inhibitory cytokine pathways for leukocyte chemoattraction and activation have been identified, but there is little insight into the operational mechanisms except for models that rely on simple receptor antagonism. We have previously identified the existence of a murine eosinophil inhibitory pathway mediated by the CXC chemokine ligand 9 (CXCL9, Mig [monokine induced by interferon-gamma]) that impressively blocks eosinophil chemoattraction and function, but the mechanism has remained elusive. We now demonstrate that Mig's inhibitory action extends beyond receptor antagonism alone. Notably, in addition to inhibiting eotaxin-induced filamentous actin (F-actin) formation and chemoattraction, Mig potently blocks platelet activating factor (PAF)- and leukotriene B4 (LTB4)-induced responses. Remarkably, Mig-treated eosinophils display an abnormal F-actin assembly in the absence of agonist stimulation. Additionally, Mig pretreatment inhibits eotaxin-induced activation of the Rho guanosine triphosphatase (GTPase) Rac, and Rac2-deficient eosinophils demonstrate an impaired transmigration and actin polymerization response to eotaxin stimulation. Furthermore, Mig was unable to inhibit eotaxin-induced responses in Rac2-deficient eosinophils. Finally, using CCR3 gene-targeted cells, Mig's inhibitory activity is demonstrated to be mediated by CC chemokine receptor 3 (CCR3). Thus, by altering agonist-induced signaling and abrogating cytoskeletal reorganization by a Rac2-dependent mechanism, Mig markedly inhibits eosinophil responses to diverse stimuli. These results establish evidence that distinct chemokines can use CCR3 to induce opposing signals in eosinophils. PMID- 15802531 TI - Glucocorticoids inhibit activation-induced cell death (AICD) via direct DNA dependent repression of the CD95 ligand gene by a glucocorticoid receptor dimer. AB - Glucocorticoids (GCs) play an important role in the regulation of peripheral T cell survival. Their molecular mechanism of action and the question of whether they have the ability to inhibit apoptosis in vivo, however, are not fully elucidated. Signal transduction through the glucocorticoid receptor (GR) is complex and involves different pathways. Therefore, we used mice with T-cell specific inactivation of the GR as well as mice with a function-selective mutation in the GR to determine the signaling mechanism. Evidence is presented for a functional role of direct binding of the GR to 2 negative glucocorticoid regulatory elements (nGREs) in the CD95 (APO-1/Fas) ligand (L) promoter. Binding of GRs to these nGREs reduces activation-induced CD95L expression in T cells. These in vitro results are fully supported by data obtained in vivo. Administration of GCs to mice leads to inhibition of activation-induced cell death (AICD). Thus, GC-mediated inhibition of CD95L expression of activated T cells might contribute to the anti-inflammatory function of steroid drugs. PMID- 15802532 TI - The FA/BRCA pathway is involved in melphalan-induced DNA interstrand cross-link repair and accounts for melphalan resistance in multiple myeloma cells. AB - Melphalan, a DNA cross-linker, is one of the most widely used and effective drugs in the treatment of multiple myeloma (MM). In this report, we demonstrate that enhanced interstrand cross-link (ICL) repair via the Fanconi anemia (FA)/BRCA pathway contributes to acquired drug resistance in melphalan-resistant myeloma cell lines, and disruption of this pathway reverses drug resistance. Using the alkaline comet assay (single-cell gel electrophoresis), we observed that melphalan-resistant cells have reduced ICL formation and enhanced ICL repair compared with melphalan-sensitive cells. Cell-cycle studies demonstrated that enhanced ICL repair released cells from melphalan-induced cell-cycle delay. Using siRNA to knock down FANCF in 8226/LR5 and U266/LR6 drug-resistant cells demonstrated a direct relationship between ICL repair capacity and drug sensitivity. Overexpression of FANCF in 8226/S and U266/S drug-sensitive cells partially reproduced the drug-resistant phenotype. These data show that enhanced DNA repair via the Fanconi anemia/BRCA pathway is involved in acquired melphalan resistance. Our findings provide for a new target to enhance response to DNA cross-linking agents in cancer treatment. PMID- 15802533 TI - The natural product honokiol induces caspase-dependent apoptosis in B-cell chronic lymphocytic leukemia (B-CLL) cells. AB - B-cell chronic lymphocytic leukemia (B-CLL) remains an incurable disease that requires innovative new approaches to improve therapeutic outcome. Honokiol is a natural product known to possess potent antineoplastic and antiangiogenic properties. We examined whether honokiol can overcome apoptotic resistance in primary tumor cells derived from B-CLL patients. Honokiol induced caspase dependent cell death in all of the B-CLL cells examined and was more toxic toward B-CLL cells than to normal mononuclear cells, suggesting greater susceptibility of the malignant cells. Honokiol-induced apoptosis was characterized by the activation of caspase-3, -8, and -9 and cleavage of poly(adenosine diphosphate ribose) polymerase (PARP). Exposure of B-CLL cells to honokiol resulted in up regulation of Bcl2-associated protein (Bax) and down-regulation of the expression of the key survival protein myeloid-cell leukemia sequence 1 (Mcl-1), which is associated with response to treatment in B-CLL patients. In addition, B-CLL cells pretreated with interleukin-4 (IL-4), a cytokine known to support B-CLL survival, underwent apoptosis when subsequently incubated with honokiol, indicating that honokiol could also overcome the prosurvival effects of IL-4. Furthermore, honokiol enhanced cytotoxicity induced by fludarabine, cladribine, or chlorambucil. These data indicate that honokiol is a potent inducer of apoptosis in B-CLL cells and should be examined for further clinical application either as a single agent or in combination with other anticancer agents. PMID- 15802534 TI - NK/iDC interaction results in IL-18 secretion by DCs at the synaptic cleft followed by NK cell activation and release of the DC maturation factor HMGB1. AB - Interaction of natural killer (NK) cells with autologous immature dendritic cells (DCs) results in reciprocal activation; however, the underlying mechanisms are so far elusive. We show here that NK cells trigger immature DCs to polarize and secrete interleukin 18 (IL-18), a cytokine lacking a secretory leader sequence. This occurs through a Ca2+-dependent and tubulin-mediated recruitment of IL-18 containing secretory lysosomes toward the adhering NK cell. Lysosome exocytosis and IL-18 secretion are restricted at the synaptic cleft, thus allowing activation of the interacting NK cells without spreading of the cytokine. In turn, DC-activated NK cells secrete the proinflammatory cytokine high mobility group B1 (HMGB1), which induces DC maturation and protects DCs from lysis. Also HMGB1 is a leaderless cytokine that undergoes regulated secretion. Differently from IL-18, soluble HMGB1 is consistently detected in NK/DC supernatants. These data point to secretion of leaderless cytokines as a key event for the reciprocal activation of NK cells and DCs. DCs initiate NK cell activation by targeted delivery of IL-18, thus instructing NK cells in the absence of adaptive-type cytokines; in turn, activated NK cells release HMGB1, which promotes inflammation and induces DC maturation, thus favoring the onset of the adaptive immune response. PMID- 15802535 TI - The LPL/ADAM29 expression ratio is a novel prognosis indicator in chronic lymphocytic leukemia. AB - Although the zeta-associated protein of 70 kDa (ZAP-70) is overexpressed in patients with chronic lymphocytic leukemia (CLL) displaying unmutated IGVH genes and poor prognosis, a previous microarray study from our group identified overexpression of LPL and ADAM29 genes among unmutated and mutated CLL, respectively. To assess the prognostic value of these genes, we quantified their expression by real-time quantitative polymerase chain reaction (PCR) in a cohort of 127 patients with CLL and correlated this with clinical outcome, IGVH mutational status, and ZAP-70 protein expression. IGVH mutational status, ZAP-70, and the LPL and ADAM29 mRNA ratios (L/A ratio) were predictive of event-free survival for the whole cohort and for patients with stage A disease. In patients in stage B and C, the L/A ratio was an independent prognostic factor, whereas ZAP 70 did not predict survival. Simultaneous usage of the L/A ratio and ZAP-70 expression allowed an almost perfect (99%) assessment of the IGVH status in the 80% of patients with concordant results (L/A+, ZAP-70+ or L/A-, ZAP-70-). LPL and ADAM29 gene expression could also be determined by a simple competitive multiplex reverse transcription PCR assay. Overall, quantification of LPL and ADAM29 gene expression is a strong prognostic indicator in CLL, providing better prognostic assessment than ZAP-70 in advanced stages of the disease. PMID- 15802536 TI - Evolving trends in qualitative research: advances in mixed-method design. PMID- 15802538 TI - Illness representations of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a chronic illness that usually follows a fluctuating course and, therefore, can be difficult to diagnose. Consequently, individuals with SLE often hold beliefs about the nature and course of the illness that are at odds with medical opinion. Clearly, a shared understanding of the illness would be beneficial to illness management. In this study, the authors used semistructured interviews and interpretative phenomenological analysis to explore the content of the illness representations held by 36 individuals diagnosed with SLE. They also identified the ways in which these illness representations were generated initially and attempted to determine whether changes occurred over time in these illness representations, as predicted by the self-regulatory model. PMID- 15802537 TI - If I didn't have HIV, I'd be dead now: illness narratives of drug users living with HIV/AIDS. AB - The purpose of this study is to illuminate the experiences of lower income, urban, HIV-positive drug users. The authors asked 60 participants about HIV risk behaviors, the impact of HIV on their lives, religious beliefs, life plans, relationships, and work-related issues both prior to and since diagnosis. They developed a theoretical framework based on illness narratives and ambiguous loss theories. Themes pertaining to both physical and emotional or spiritual dimensions were located within Benefit, Loss, or Status Quo orientations. The findings contribute to researchers' understanding of the HIV/AIDS illness experiences among the very marginalized and have important implications for physical and mental health care professionals working with HIV-positive drug users. PMID- 15802539 TI - The incidental discovery of nonpaternity through genetic carrier screening: an exploration of lay attitudes. AB - With advances in genetic medicine, paternity is increasingly being determined by genetic rather than social markers. In this article, the author examines the complex nature of paternity and the way in which it has been constructed socially, legally, and medically. She then presents the results from an empirical study on public attitudes toward genetic testing and paternity. In particular, she examines lay attitudes toward an ethical dilemma posed by the incidental discovery of nonpaternity during the process of genetic testing for a recessively inherited disease. She concludes the article by drawing attention to the potential problems inherent in the intervention of genetic medicine into family relationships. PMID- 15802540 TI - The experience of interferon-based treatments for hepatitis C infection. AB - Clinical trials of interferon-based treatments for hepatitis C infection show decrements in patients' health-related quality of life due to side effects of therapy. The impact of side effects on patients' overall quality of life still remains unclear. To explore this issue, the authors interviewed people living in New South Wales, Australia, who had undergone treatment for hepatitis C. Their aim in this article is to report participants' experiences of treatment side effects. In Australia, this information is important, because a new interferon based regimen has been adopted as the mainstay of hepatitis C treatment, and it is predicted that many more people will seek treatment. The authors argue for further qualitative research to enhance knowledge of the impact of this therapy. PMID- 15802541 TI - Objectivity and subjectivity in the ethnographic method. AB - The author discusses the application of the concepts of objectivity and subjectivity in ethnographic theory and research, and finds that one kind of subjectivity, that of applying a particular perspective to ethnography, is central and inevitable. Today, we acknowledge that objectivity is relative to a given perspective or preunderstanding, but the applied perspective must compete with other perspectives or paradigms in its effectiveness in our understanding and managing of a lived reality. In some situations, where no such fruitful alternative exists, a simple correspondence between word and world can be an acceptable notion of truth. Here, an all-encompassing consensus of what would be objective and subjective can be expected. Thus, the concepts retain a central function without completely altering the meanings they have in everyday and scientific language, although the values traditionally ascribed to the notions have changed to some extent and a more refined conceptualization has been reached. PMID- 15802543 TI - Cooperation or co-optation?: Assessing the methodological benefits and barriers involved in conducting qualitative research through medical institutional settings. AB - In this article, the authors highlight some benefits of and barriers to doing qualitative research in association with hospital-based services. They first describe an ongoing qualitative research project that involves interviewing women about their post-sexual assault medicolegal experiences in hospital-situated sexual assault centers across a large Canadian province. Their methodological journey led them to engage program coordinators at these centers to assist with locating participants and qualified interviewers, and with negotiating the demands of their respective research ethics boards. They outline the ways in which their project was shaped, positively and negatively, by working with them in medical institutions. They conclude by recommending that hospitals and hospital ethics boards counteract tendencies toward paternalism by recognizing the value of feminist qualitative research contributions to the activities of their own sexual assault centers and to the recovery of sexually assaulted women. Such recognition might be productively engaged by adopting an ethics-in-process approach. PMID- 15802542 TI - Case study research: the view from complexity science. AB - Many wonder why there has been so little change in care quality despite substantial quality improvement efforts. Questioning why current approaches are not making true changes draws attention to the organization as a source of answers. The authors bring together the case study method and complexity science to suggest new ways to study health care organizations. The case study provides a method for studying systems. Complexity theory suggests that keys to understanding the system are contained in patterns of relationships and interactions among the system's agents. They propose some of the "objects" of study that are implicated by complexity theory and discuss how studying these using case methods might provide useful maps of the system. They offer complexity theory, partnered with case study method, as a place to begin the daunting task of studying a system as an integrated whole. PMID- 15802544 TI - The problems with interviews. AB - Despite the popularity of the interview in qualitative research, methodological and theoretical problems remain. In this article, the author critically examines some of these problems for the researcher. He deals with the problems of power and resistance, distinguishing truth from authenticity, the (im)possibility of consent if knowing is a problem for both the interviewer and the interviewee, and the nature and significance of stories and the self. Although it is not always possible to address these problems directly, the author seeks in this article to create a dialogue with all of us for whom the interview is judged to be the appropriate answer to the research question "How can I know...?" PMID- 15802545 TI - Use of an audit in violence prevention research. AB - Auditing is an effective tool for articulating the trustworthiness and credibility of qualitative research. However, little information exists on how to conduct an audit. In this article, the authors illustrate their use of an audit team to explore the methods and preliminary findings of a study aimed at identifying the relevant and challenging problems experienced by urban teenagers. This study was the first in a series of studies to improve the ecological validity of violence prevention programs for high-risk urban teenagers, titled Identifying Essential Skills for Violence Prevention. The five phases of this audit were engaging the auditor, becoming familiar with the study, discussing methods and determining strengths and limitations, articulating audit findings, and planning subsequent research. Positioning the audit before producing final results allows researchers to address many study limitations, uncover potential sources of bias in the thematic structure, and systematically plan subsequent steps in an emerging design. PMID- 15802546 TI - Lighten up: the citation dilemma in qualitative research. AB - The balance between what we learn from research and what others have said about similar topics can pose dilemmas for qualitative researchers. How many citations are enough? How much allegiance do we owe to other scholars and how much to informants' lived experiences, our theoretical analyses, and the lessons we draw about methods and methodologies from our own experience as researchers? The various reasons we use or do not use citations in writing up results and writing about methods are the topics of this article. PMID- 15802547 TI - HFE mutations in the pathobiology of hemophilic arthropathy. PMID- 15802548 TI - Hyperhomocysteinemia: cause or effect of disease? PMID- 15802549 TI - Use of desmopressin (DDAVP) during early pregnancy in factor VIII-deficient women. PMID- 15802550 TI - Serum C-reactive protein parallels secretory phospholipase A2 in sickle cell disease patients with vasoocclusive crisis or acute chest syndrome. PMID- 15802552 TI - Embryonic lung side population cells are hematopoietic and vascular precursors. AB - Side population (SP) cells are a select cell population identified by a capacity to efflux Hoechst dye that are highly enriched for stem/progenitor cell activity. In this study, we found that SP cells comprised of CD45(+) and CD45(-) subtypes are present in the embryonic lung (E-SP) at levels varying with gestational age. Long-term in vivo competitive blood reconstitution studies demonstrated that hematopoeitic stem cell capacity resided within the CD45(+) E-SP cell subset. Immunophenotyping of CD45(-) E-SP cells determined that this population consists of two subtypes: CD31(-) and CD31(+). Limited gene expression profiling indicated that CD45(-)/CD31(-) E-SP cells have features of smooth muscle precursors, and give rise to smooth muscle in culture. On the other hand, CD45(-)/CD31(+) E-SP cells express genes characteristic of endothelium, but by themselves do not grow or differentiate in culture. Co-culture of CD45(-)/CD31(+) and CD45(-)/CD31(-) E SP cells, however, resulted in the formation of complex tubular networks that express markers of endothelium. Together, these findings illustrate that embryonic lung SP cells are heterogeneous, composed of hematopoeitic and nonhematopoeitic progenitors, and may play a key role in the formation of the lung vasculature. PMID- 15802551 TI - Regulation of interleukin-5-induced beta2-integrin adhesion of human eosinophils by phosphoinositide 3-kinase. AB - We examined the role of phosphoinositide 3-kinase (PI3K) in integrin-mediated eosinophil adhesion. Deltap85, a dominant-negative form of the class IA PI3K adaptor subunit, was fused to an HIV-TAT protein transduction domain (TAT Deltap85). Recombinant TAT-Deltap85 inhibited interleukin (IL)-5-stimulated phosphorylation of protein kinase B, a downstream target of PI3K. beta(2) Integrin-dependent adhesion caused by IL-5 to the plated intracellular adhesion molecule-1 surrogate, bovine serum albumin, was inhibited by TAT-Deltap85 in a concentration-dependent manner. Similarly, two PI3K inhibitors, wortmannin and LY294002, blocked eosinophil adhesion to plated bovine serum albumin. By contrast, beta(1)-integrin-mediated eosinophil adhesion to vascular cell adhesion moelcule-1 was not blocked by TAT-Deltap85, wortmannin, or LY294002. Rottlerin, a protein kinase C (PKC)-delta inhibitor, also blocked beta(2)-integrin adhesion of eosinophils caused by IL-5, whereas beta(1) adhesion to vascular cell adhesion molecule-1 was not affected. IL-5 caused translocation of PKCdelta from the cytosol to cell membrane; inhibition of PI3K by wortmannin blocked translocation of PKCdelta. Western blot analysis demonstrated that extracellular signal regulated kinase phosphorylation, a critical intermediary in adhesion elicited by IL-5, was blocked by inhibition of either PI3K or PKC-delta. These data suggest that extracellular signal-regulated kinase-mediated adhesion of beta(2)-integrin caused by IL-5 is mediated in human eosinophils by a class IA PI3K through activation of a PKCdelta pathway. PMID- 15802553 TI - alpha-Galactosylceramide, a ligand of natural killer T cells, inhibits allergic airway inflammation. AB - alpha-Galactosylceramide (alpha-GalCer) is a specific ligand of natural killer T cells (NKT cells) that regulates the immune responses such as tumor rejection and autoimmunity by producing interferon (IFN)-gamma and interleukin (IL)-4. However, it has not been determined whether alpha-GalCer-activated NKT cells modulate allergic inflammation. Because alpha-GalCer induces a large amount of IFN-gamma production by NKT cells, we hypothesized that an in vivo administration of alpha GalCer could inhibit allergic airway inflammation in mice. Strikingly, a single intraperitoneal injection of alpha-GalCer almost completely abrogated an infiltrate with eosinophils in the lung tissue as well as in the bronchoalveolar lavage. This inhibition of allergic inflammation was associated with a significant decrease in the levels of IL-4, IL-5, and IL-13 in bronchoalveolar lavage fluid and in the number of goblet cells. In addition, this ligand significantly inhibited airway hyperresponsiveness to inhaled methacholine and raised the serum levels of ovalbumin-specific IgG2a with a decrease in those of ovalbumin-specific IgE. In IFN-gamma knockout mice, however, alpha-GalCer failed to exert such inhibitory effects in this asthma model. These results indicate that alpha-GalCer prevents allergic airway inflammation possibly through IFN gamma production by ligand-activated NKT cells, suggesting the potential therapeutic application of alpha-GalCer in asthma. PMID- 15802554 TI - Activation of 5-HT1A receptors in the medullary raphe reduces cardiovascular changes elicited by acute psychological and inflammatory stresses in rabbits. AB - The present strategy for the prevention of excessive sympathetic neural traffic to the heart relies on the use of beta-blockers, drugs that act at the heart end of the brain-heart axis. In the present study, we attempted to suppress cardiac sympathetic nerve activity by affecting the relevant cardiomotoneurons in the brain using the selective serotonin-1A (5-HT(1A)) receptor agonist 8-hydroxy-2 (di-n-propylamino)tetralin (8-OH-DPAT). In conscious, unrestrained rabbits, instrumented for recordings of heart rate, arterial pressure, or cardiac output, we provoked increases in cardiac sympathetic activity by psychological (loud sound, pinprick, and air jet) or inflammatory (0.5 microg/kg iv lipopolysaccharide) stresses. Pinprick and air-jet stresses elicited transient increases in heart rate (+50 +/- 7 and +38 +/- 4 beats/min, respectively) and in mean arterial pressure (+16 +/- 2 and +15 +/- 3 mmHg, respectively). Lipopolysaccharide injection caused sustained increases in heart rate (from 210 +/- 3 to 268 +/- 10 beats/min) and in arterial pressure (from 74 +/- 3 to 92 +/- 4 mmHg). Systemically administered 8-OH-DPAT (0.004-0.1 mg/kg) substantially attenuated these responses in a dose-dependent manner. Drug effects were prevented by a selective 5-HT(1A) receptor antagonist, WAY-100635 (0.1 mg/kg iv). Similarly to systemic administration, microinjection of 8-OH-DPAT (500 nl of 10 mM solution) into the medullary raphe-parapyramidal region caused antitachycardic effects during stressful stimulation and during lipopolysaccharide-elicited tachycardia. This is the first demonstration that activation of 5-HT(1A) receptors in the medullary raphe-parapyramidal area causes suppression of neurally mediated cardiovascular changes during acute psychological and immune stresses. PMID- 15802555 TI - Specific carotid body chemostimulation is sufficient to elicit phrenic poststimulus frequency decline in a novel in situ dual-perfused rat preparation. AB - Time-dependent ventilatory responses to hypoxic and hypercapnic challenges, such as posthypoxic frequency decline (PHxFD) and posthypercapnic frequency decline (PHcFD), could profoundly affect breathing stability. However, little is known about the mechanisms that mediate these phenomena. To determine the contribution of specific carotid body chemostimuli to PHxFD and PHcFD, we developed a novel in situ arterially perfused, vagotomized, decerebrate rat preparation in which central and peripheral chemoreceptors are perfused separately (i.e., a nonanesthetized in situ dual perfused preparation). We confirmed that 1) the perfusion of central and peripheral chemoreceptor compartments was independent by applying specific carotid body hypoxia and hypercapnia before and after carotid sinus nerve transection, 2) the PCO(2) chemoresponse of the dual perfused preparation was similar to other decerebrate preparations, and 3) the phrenic output was stable enough to allow investigation of time-dependent phenomena. We then applied four 5-min bouts (separated by 5 min) of specific carotid body hypoxia (40 Torr PO(2) and 40 Torr PCO(2)) or hypercapnia (100 Torr PO(2) and 60 Torr PCO(2)) while holding the brain stem PO(2) and PCO(2) constant. We report the novel finding that specific carotid body chemostimuli were sufficient to elicit several phrenic time-dependent phenomena in the rat. Hypoxic challenges elicited PHxFD that increased with bout, leading to progressive augmentation of the phrenic response. Conversely, hypercapnia elicited short-term depression and PHcFD, neither of which was bout dependent. These results, placed in the context of previous findings, suggest multiple physiological mechanisms are responsible for PHxFD and PHcFD, a redundancy that may illustrate that these phenomena have significant adaptive advantages. PMID- 15802557 TI - Quantitative analysis of voltage-gated potassium currents from primary equine (Equus caballus) and elephant (Loxodonta africana) articular chondrocytes. AB - In this comparative study, we have established in vitro models of equine and elephant articular chondrocytes, examined their basic morphology, and characterized the biophysical properties of their primary voltage-gated potassium channel (Kv) currents. Using whole cell patch-clamp electrophysiological recording from first-expansion and first-passage cells, we measured a maximum Kv conductance of 0.15 +/- 0.04 pS/pF (n = 10) in equine chondrocytes, whereas that in elephant chondrocytes was significantly larger (0.8 +/- 0.4 pS/pF, n = 4, P 0.05). Preformed MMP 2 and MMP-9 were probably activated in CHF, because neither their gene expression nor protein levels were altered (P > 0.05). Serum concentrations of TNF-alpha and monocyte chemoattractant protein-1 remained unchanged (P > 0.05) between CHF and Sham rats during the 6-wk observation period. We conclude that development of CHF in rats enhances MMP activity, which in turn may distort the normal contractile function of skeletal muscle, thereby contributing to increased skeletal muscle fatigue. PMID- 15802563 TI - Hypothalamic-pituitary disconnection in fetal sheep blocks the peripartum increases in adrenal responsiveness and adrenal ACTH receptor expression. AB - Although it has been recognized for over a decade that hypothalamic-pituitary disconnection (HPD) in fetal sheep prevents the late gestation rise in plasma cortisol concentrations, the underlying mechanisms remain unclear. We hypothesized that reductions in adrenal responsiveness and ACTH receptor (ACTH-R) expression may be mediating factors. HPD or sham surgery was performed at 120 days of gestation, and catheters were placed for blood sampling. At approximately 138 days of gestation, fetuses were killed, and adrenals were removed for cell culture and analyses of ACTH-R mRNA and protein. After 48 h, adrenocortical cells were stimulated with ACTH for 2 h, and the medium was collected for cortisol measurement. The same cells were incubated overnight with medium or medium containing ACTH or forskolin (FSK), followed by ACTH stimulation (as above) and cortisol and cellular ACTH-R mRNA analyses. HPD prevented the late gestation increase in plasma cortisol and bioactive ACTH and reduced adrenal ACTH-R mRNA and protein levels by over 35%. HPD cells secreted significantly less cortisol than sham cells (3.2 +/- 1.2 vs. 47.3 +/- 11.1 ng.ml(-1).2 h(-1)) after the initial ACTH stimulation. Overnight incubation of HPD cells with ACTH or FSK restored cortisol responses to acute stimulation to levels seen in sham cells initially. ACTH-R mRNA levels in cells isolated from HPD fetuses were decreased by over 60%, whereas overnight incubation with ACTH or FSK increased levels by approximately twofold. Our findings indicate that the absence of the cortisol surge in HPD fetuses is a consequence, at least in part, of decreased ACTH-R expression and adrenal responsiveness. PMID- 15802564 TI - The kinase domain of titin controls muscle gene expression and protein turnover. AB - The giant sarcomeric protein titin contains a protein kinase domain (TK) ideally positioned to sense mechanical load. We identified a signaling complex where TK interacts with the zinc-finger protein nbr1 through a mechanically inducible conformation. Nbr1 targets the ubiquitin-associated p62/SQSTM1 to sarcomeres, and p62 in turn interacts with MuRF2, a muscle-specific RING-B-box E3 ligase and ligand of the transactivation domain of the serum response transcription factor (SRF). Nuclear translocation of MuRF2 was induced by mechanical inactivity and caused reduction of nuclear SRF and repression of transcription. A human mutation in the titin protein kinase domain causes hereditary muscle disease by disrupting this pathway. PMID- 15802565 TI - Structure of the rotor of the V-Type Na+-ATPase from Enterococcus hirae. AB - The membrane rotor ring from the vacuolar-type (V-type) sodium ion-pumping adenosine triphosphatase (Na+-ATPase) from Enterococcus hirae consists of 10 NtpK subunits, which are homologs of the 16-kilodalton and 8-kilodalton proteolipids found in other V-ATPases and in F1Fo- or F-ATPases, respectively. Each NtpK subunit has four transmembrane alpha helices, with a sodium ion bound between helices 2 and 4 at a site buried deeply in the membrane that includes the essential residue glutamate-139. This site is probably connected to the membrane surface by two half-channels in subunit NtpI, against which the ring rotates. Symmetry mismatch between the rotor and catalytic domains appears to be an intrinsic feature of both V- and F-ATPases. PMID- 15802566 TI - Human Mpp11 J protein: ribosome-tethered molecular chaperones are ubiquitous. AB - The existence of specialized molecular chaperones that interact directly with ribosomes is well established in microorganisms. Such proteins bind polypeptides exiting the ribosomal tunnel and provide a physical link between translation and protein folding. We report that ribosome-associated molecular chaperones have been maintained throughout eukaryotic evolution, as illustrated by Mpp11, the human ortholog of the yeast ribosome-associated J protein Zuo. When expressed in yeast, Mpp11 partially substituted for Zuo by partnering with the multipurpose Hsp70 Ssa, the homolog of mammalian Hsc70. We propose that in metazoans, ribosome associated Mpp11 recruits the multifunctional soluble Hsc70 to nascent polypeptide chains as they exit the ribosome. PMID- 15802567 TI - An octane-fueled solid oxide fuel cell. AB - There are substantial barriers to the introduction of hydrogen fuel cells for transportation, including the high cost of fuel-cell systems, the current lack of a hydrogen infrastructure, and the relatively low fuel efficiency when using hydrogen produced from hydrocarbons. Here, we describe a solid oxide fuel cell that combines a catalyst layer with a conventional anode, allowing internal reforming of iso-octane without coking and yielding stable power densities of 0.3 to 0.6 watts per square centimeter. This approach is potentially the basis of a simple low-cost system that can provide substantially higher fuel efficiency by using excess fuel-cell heat for the endothermic reforming reaction. PMID- 15802568 TI - All Fools' Day. PMID- 15802569 TI - U.K. bioethics. Divided committee urges less restriction on embryo research. PMID- 15802570 TI - Select agents. Researchers relieved by final biosecurity rules. PMID- 15802572 TI - Space exploration. Japan weighs moon and beyond. PMID- 15802573 TI - French science. Politician sails into a storm at oceans agency. PMID- 15802571 TI - Infectious diseases. A puzzling outbreak of Marburg disease. PMID- 15802574 TI - Italian research. Universities and institutes face industrial revolution. PMID- 15802576 TI - Mathematics. 'Cranky' proof reveals hidden regularities. PMID- 15802575 TI - Neuroscience. Economic game shows how the brain builds trust. PMID- 15802577 TI - Academic jobs. Tenured UCLA professor under fire. PMID- 15802578 TI - Physics. High-energy physics: Exit America? PMID- 15802579 TI - Ecology. Ecosystem services. PMID- 15802580 TI - Ecology. Choose your own world. PMID- 15802581 TI - American Chemical Society meeting. Unnatural amino acid could prove boon for protein therapeutics. PMID- 15802582 TI - American Chemical Society meeting. Nanofibers seed blood vessels. PMID- 15802583 TI - American Chemical Society meeting. Fast, sensitive scan targets anthrax. PMID- 15802585 TI - Don't call them co-eds! PMID- 15802584 TI - NIH response to open letter. PMID- 15802586 TI - Comment on "Oscillations in NF-kappaB signaling control the dynamics of gene expression". PMID- 15802588 TI - Global voices of science. When science is not enough: fighting genetic disease in Brazil. PMID- 15802589 TI - Physiology. Walking made simple. PMID- 15802590 TI - Geology. The calibration of Ediacaran time. PMID- 15802591 TI - Evolution. Where we're hot, they're not. PMID- 15802592 TI - Psychology. Beyond a joke: from animal laughter to human joy? PMID- 15802593 TI - Materials science. Playing nature's game with artificial muscles. PMID- 15802594 TI - Cell biology. Kinasing and clipping down the NF-kappa B trail. PMID- 15802596 TI - Abundance of cellular material and proteins in the atmosphere. AB - Suspended atmospheric particles play a crucial role in any global climate scenario: They can both enforce and suppress radiative forcing. In developing climate modeling further, a deeper understanding of atmospheric aerosol is needed. Because of extreme local and temporal variations, proper incorporation of aerosols into models requires modeling of the aerosol itself. It turns out that cellular material and proteins compose up to 25% of the atmospheric aerosol. Consequently, the source strength of the biogenic aerosol in general must be corrected and should be estimated on the order of other major aerosol sources. PMID- 15802595 TI - Global iron connections between desert dust, ocean biogeochemistry, and climate. AB - The environmental conditions of Earth, including the climate, are determined by physical, chemical, biological, and human interactions that transform and transport materials and energy. This is the "Earth system": a highly complex entity characterized by multiple nonlinear responses and thresholds, with linkages between disparate components. One important part of this system is the iron cycle, in which iron-containing soil dust is transported from land through the atmosphere to the oceans, affecting ocean biogeochemistry and hence having feedback effects on climate and dust production. Here we review the key components of this cycle, identifying critical uncertainties and priorities for future research. PMID- 15802597 TI - Role of marine biology in glacial-interglacial CO2 cycles. AB - It has been hypothesized that changes in the marine biological pump caused a major portion of the glacial reduction of atmospheric carbon dioxide by 80 to 100 parts per million through increased iron fertilization of marine plankton, increased ocean nutrient content or utilization, or shifts in dominant plankton types. We analyze sedimentary records of marine productivity at the peak and the middle of the last glacial cycle and show that neither changes in nutrient utilization in the Southern Ocean nor shifts in plankton dominance explain the CO2 drawdown. Iron fertilization and associated mechanisms can be responsible for no more than half the observed drawdown. PMID- 15802598 TI - Getting to know you: reputation and trust in a two-person economic exchange. AB - Using a multiround version of an economic exchange (trust game), we report that reciprocity expressed by one player strongly predicts future trust expressed by their partner-a behavioral finding mirrored by neural responses in the dorsal striatum. Here, analyses within and between brains revealed two signals-one encoded by response magnitude, and the other by response timing. Response magnitude correlated with the "intention to trust" on the next play of the game, and the peak of these "intention to trust" responses shifted its time of occurrence by 14 seconds as player reputations developed. This temporal transfer resembles a similar shift of reward prediction errors common to reinforcement learning models, but in the context of a social exchange. These data extend previous model-based functional magnetic resonance imaging studies into the social domain and broaden our view of the spectrum of functions implemented by the dorsal striatum. PMID- 15802599 TI - Spin-charge separation and localization in one dimension. AB - We report on measurements of quantum many-body modes in ballistic wires and their dependence on Coulomb interactions, obtained by tunneling between two parallel wires in an GaAs/AlGaAs heterostructure while varying electron density. We observed two spin modes and one charge mode of the coupled wires and mapped the dispersion velocities of the modes down to a critical density, at which spontaneous localization was observed. Theoretical calculations of the charge velocity agree well with the data, although they also predict an additional charge mode that was not observed. The measured spin velocity was smaller than theoretically predicted. PMID- 15802600 TI - Simultaneous tomography and diffraction analysis of creep damage. AB - Creep damage by void nucleation and growth limits the lifetime of components subjected to loading at high temperatures. We report a combined tomography and diffraction experiment using high-energy synchrotron radiation that permitted us to follow in situ void growth and microstructure development in bulk samples. The results reveal that void growth versus time follows an exponential growth law. The formation of large void volumes coincides with texture evolution and dislocation density, reaching a steady state. Creep damage during a large proportion of sample creep life is homogeneous before damage localization occurs, which leads to rapid failure. The in situ determination of void evolution in bulk samples should allow for the assessment of creep damage in metallic materials and subsequently for lifetime predictions about samples and components that are subject to high-temperature loading. PMID- 15802601 TI - Mortality and greenhouse gas impacts of biomass and petroleum energy futures in Africa. AB - We analyzed the mortality impacts and greenhouse gas (GHG) emissions produced by household energy use in Africa. Under a business-as-usual (BAU) scenario, household indoor air pollution will cause an estimated 9.8 million premature deaths by the year 2030. Gradual and rapid transitions to charcoal would delay 1.0 million and 2.8 million deaths, respectively; similar transitions to petroleum fuels would delay 1.3 million and 3.7 million deaths. Cumulative BAU GHG emissions will be 6.7 billion tons of carbon by 2050, which is 5.6% of Africa's total emissions. Large shifts to the use of fossil fuels would reduce GHG emissions by 1 to 10%. Charcoal-intensive future scenarios using current practices increase emissions by 140 to 190%; the increase can be reduced to 5 to 36% using currently available technologies for sustainable production or potentially reduced even more with investment in technological innovation. PMID- 15802602 TI - A Late Jurassic digging mammal and early mammalian diversification. AB - A fossil mammal from the Late Jurassic Morrison Formation, Colorado, has highly specialized teeth similar to those of xenarthran and tubulidentate placental mammals and different from the generalized insectivorous or omnivorous dentitions of other Jurassic mammals. It has many forelimb features specialized for digging, and its lumbar vertebrae show xenarthrous articulations. Parsimony analysis suggests that this fossil represents a separate basal mammalian lineage with some dental and vertebral convergences to those of modern xenarthran placentals, and reveals a previously unknown ecomorph of early mammals. PMID- 15802603 TI - Neuronal coherence as a mechanism of effective corticospinal interaction. AB - Neuronal groups can interact with each other even if they are widely separated. One group might modulate its firing rate or its internal oscillatory synchronization to influence another group. We propose that coherence between two neuronal groups is a mechanism of efficient interaction, because it renders mutual input optimally timed and thereby maximally effective. Modulations of subjects' readiness to respond in a simple reaction-time task were closely correlated with the strength of gamma-band (40 to 70 hertz) coherence between motor cortex and spinal cord neurons. This coherence may contribute to an effective corticospinal interaction and shortened reaction times. PMID- 15802604 TI - PDK1 nucleates T cell receptor-induced signaling complex for NF-kappaB activation. AB - Activation of the transcription factor NF-kappaB after engagement of the T cell receptor (TCR) is important for T cell proliferation and activation during the adaptive immune response. Recent reports have elucidated a signaling pathway that involves the protein kinase C (PKC), the scaffold protein CARD11 (also called CARMA-1), the caspase recruitment domain (CARD)-containing protein Bcl10, and the paracaspase (protease related to caspases) MALT1 as critical intermediates linking the TCR to the IkappaB kinase (IKK) complex. However, the events proximal to the TCR that initiate the activation of this signaling pathway remain poorly defined. We demonstrate that 3-phosphoinositide-dependent kinase 1 (PDK1) has an essential role in this pathway by regulating the activation of PKC and through signal-dependent recruiting of both PKC and CARD11 to lipid rafts. PDK1 associated PKC recruits the IKK complex, whereas PDK1-associated CARD11 recruits the Bcl10-MALT1 complex, thereby allowing activation of the IKK complex through Bcl10-MALT1-dependent ubiquitination of the IKK complex subunit known as NEMO (NF kappaB essential modifier). Hence, PDK1 plays a critical role by nucleating the TCR-induced NF-kappaB activation pathway in T cells. PMID- 15802606 TI - Brief report: psychosocial adjustment in adolescents with inflammatory bowel disease. AB - BACKGROUND: Inflammatory bowel disease (IBD) is an ideal disease for investigating adolescent adjustment to chronic illness, given its embarrassing, socially limiting, appearance-changing symptoms and adolescent onset. OBJECTIVE: To compare psychosocial adjustment among adolescents with a chronic illness to that of healthy adolescents and examine the role of adolescent disease onset. METHODS: Participants were 50 adolescents with IBD and their parents, and parents of 42 healthy comparison adolescents who completed questionnaires assessing behavioral, emotional, social, and family functioning. RESULTS: Adolescents with IBD were reported to have worse anxious and/or depressed and social problems than healthy adolescents. More adolescents with IBD were reported to have clinically significant social problems. Those diagnosed during adolescence were reported to have significantly worse social competence scores. CONCLUSIONS: Adolescents with a chronic illness such as IBD may be at higher risk for specific psychosocial difficulties than healthy adolescents. Diagnosis of a chronic illness during adolescence may have implications for social functioning. PMID- 15802605 TI - Translational operator of mRNA on the ribosome: how repressor proteins exclude ribosome binding. AB - The ribosome of Thermus thermophilus was cocrystallized with initiator transfer RNA (tRNA) and a structured messenger RNA (mRNA) carrying a translational operator. The path of the mRNA was defined at 5.5 angstroms resolution by comparing it with either the crystal structure of the same ribosomal complex lacking mRNA or with an unstructured mRNA. A precise ribosomal environment positions the operator stem-loop structure perpendicular to the surface of the ribosome on the platform of the 30S subunit. The binding of the operator and of the initiator tRNA occurs on the ribosome with an unoccupied tRNA exit site, which is expected for an initiation complex. The positioning of the regulatory domain of the operator relative to the ribosome elucidates the molecular mechanism by which the bound repressor switches off translation. Our data suggest a general way in which mRNA control elements must be placed on the ribosome to perform their regulatory task. PMID- 15802607 TI - The relationship of prenatal alcohol exposure and the postnatal environment to child depressive symptoms. AB - OBJECTIVE: This study examined the association between prenatal alcohol exposure and child depressive symptoms, and the mediating effects of maternal and child characteristics. METHODS: Participants were 42 children aged 4-5 years and their biological mothers. Prenatal alcohol consumption was assessed by self-report of maximum drinks per drinking occasion. The Pictorial Depression Scale (PDS) measured child depressive symptoms. Mother-child interactions were assessed using the family interaction puzzle task. RESULTS: Structural equation modeling indicated that prenatal alcohol exposure was associated with more negative child affect. In turn, mothers of more negative children were less emotionally connected to their children, and those children had higher levels of depressive symptomatology. Results could not be explained by current maternal drinking patterns or maternal depression. CONCLUSIONS: Study findings highlight the importance of examining prenatal alcohol exposure as a risk factor in the prediction of childhood-onset depression and the environmental mechanisms that may mediate that relationship. PMID- 15802609 TI - Vaccines against bacterial meningitis. AB - Meningitis remains an important cause of morbidity and mortality among children >5 years of age and is especially prevalent in developing countries. Effective routine immunization against Hib, pneumococcus and serogroupC meningococcus has had a significant impact on both invasive disease and carriage caused by these encapsulated bacteria. The major challenge in prevention of meningitis remains the delivery of vaccines worldwide, especially to resource-poor regions with the greatest disease burden. PMID- 15802608 TI - Mental health outcomes of cocaine-exposed children at 6 years of age. AB - OBJECTIVE: To assess 6-year-old cocaine- and noncocaine-exposed children's mental health outcomes controlling for potential confounders. METHODS: The sample consisted of 322 children [169 cocaine exposed (CE) and 153 noncocaine exposed (NCE)] enrolled in a longitudinal study since birth. At age 6, children were assessed for mental health symptoms using the Dominic Interactive (DI), a child self-report measure, and the Child Behavior Checklist (CBCL), a caregiver report of behavioral problems. RESULTS: CE children were more likely to self-report symptoms in the probable clinical range for oppositional defiant disorder (ODD) and attention deficit hyperactivity disorder (ADHD). In contrast, prenatal cocaine exposure was not related to child behavior based on the CBCL. After control for exposure, CE children in adoptive or foster care were rated as having more problems with aggression, externalizing behaviors, and total behavioral problems than NCE children and CE children in maternal or relative care. Also, CE children in adoptive or foster care self-reported more externalizing symptoms than CE children in maternal or relative care and NCE children. Findings could not be attributed to caregiver intelligence or depressive symptoms, or to the quality of the home environment. CONCLUSIONS: CE children report more symptoms of ODD and ADHD than nonexposed children. Adoptive or foster caregivers rated their CE children as having more behavioral problems than did maternal or relative caregivers of CE children or parents of NCE children. Although further studies are needed to understand the basis for the more negative ratings by adoptive or foster caregivers of their CE children, the self-report of CE children indicates a need for psychological interventions. PMID- 15802610 TI - Non-invasive ventilation for acute respiratory disease. AB - Non-invasive ventilation (NIV) has been shown to be effective in acute respiratory failure of various aetiologies in different health care systems and ward settings. It should be seen as complementary to invasive ventilation and primarily a means of preventing some patients from deteriorating to the point at which intubation is needed. Generally it is best initiated early before assisted ventilation is mandatory, although it has been shown to be effective even in very sick patients. Important benefits include the avoidance of endotracheal-tube associated infections, which carry an important morbidity and mortality, and a reduction in health care costs. The most important ingredient for an acute NIV service is a well-trained enthusiastic ward team. PMID- 15802611 TI - Ric-8 enhances G protein betagamma-dependent signaling in response to betagamma binding peptides in intact cells. AB - Peptides derived from a random-peptide phage display screen with purified Gbeta(1)gamma(2) subunits as the target promote the dissociation of G protein heterotrimers in vitro and activate G protein signaling in intact cells. In vitro, one of these peptides (SIRKALNILGYPDYD; SIRK) promotes subunit dissociation by binding directly to Gbetagamma subunits and accelerating the dissociation of GalphaGDP without catalyzing nucleotide exchange. The experiments described here were designed to test whether the mechanism of SIRK action in vitro is in fact the mechanism of action in intact cells. We created a mutant of Gbeta(1) subunits (beta(1)W332A) that does not bind SIRK in vitro. Transfection of Gbeta(1)W332A mutant into Chinese hamster ovary cells blocked peptide-mediated activation of extracellular signal-regulated kinase (ERK), but it did not affect receptor-mediated Gbetagamma subunit-dependent ERK activation, indicating that Gbetagamma subunits are in fact the direct target in cells responsible for ERK activation. To determine whether free Galpha subunits were released from G protein heterotrimers upon peptide treatment, cells were transfected with Ric-8A, a guanine nucleotide exchange factor for free GalphaGDP, but not heterotrimeric G proteins. Ric-8A-transfected cells displayed enhanced myristoyl-SIRKALNILGYPDYD (mSIRK)-dependent inositol phosphate (IP) release and ERK activation. Ric-8A also enhanced ERK activation by the G(i)-linked G protein coupled receptor agonist lysophosphatidic acid. Inhibitors of Gbetagamma subunit function blocked Ric-8 enhanced activation of ERK and IP release. These results suggest that one potential function of Ric-8 in cells is to enhance G protein Gbetagamma subunit signaling. Overall, these experiments provide further support for the hypothesis that mSIRK promotes G protein subunit dissociation to release free betagamma subunits in intact cells. PMID- 15802612 TI - Interdependency of beta-adrenergic receptors and CFTR in regulation of alveolar active Na+ transport. AB - Beta-adrenergic receptors (betaAR) regulate active Na+ transport in the alveolar epithelium and accelerate clearance of excess airspace fluid. Accumulating data indicates that the cystic fibrosis transmembrane conductance regulator (CFTR) is important for upregulation of the active ion transport that is needed to maintain alveolar fluid homeostasis during pulmonary edema. We hypothesized that betaAR regulation of alveolar active transport may be mediated via a CFTR dependent pathway. To test this hypothesis we used a recombinant adenovirus that expresses a human CFTR cDNA (adCFTR) to increase CFTR function in the alveolar epithelium of normal rats and mice. Alveolar fluid clearance (AFC), an index of alveolar active Na+ transport, was 92% greater in CFTR overexpressing lungs than controls. Addition of the Cl- channel blockers NPPB, glibenclamide, or bumetanide and experiments using Cl- free alveolar instillate solutions indicate that the accelerated AFC in this model is due to increased Cl- channel function. Conversely, CFTR overexpression in mice with no beta1- or beta2-adrenergic receptors had no effect on AFC. Overexpression of a human beta2AR in the alveolar epithelium significantly increased AFC in normal mice but had no effect in mice with a non-functional human CFTR gene (Deltaphi508 mutation). These studies indicate that upregulation of alveolar CFTR function speeds clearance of excess fluid from the airspace and that CFTRs effect on active Na+ transport requires the betaAR. These studies reveal a previously undetected interdependency between CFTR and betaAR that is essential for upregulation of active Na+ transport and fluid clearance in the alveolus. PMID- 15802613 TI - Adenosine-dependent induction of glutathione peroxidase 1 in human primary endothelial cells and protection against oxidative stress. AB - Cellular glutathione peroxidase (GPx-1), a selenocysteine-containing enzyme, plays a central role in protecting cells from oxidative injury. GPx-1 is ubiquitously expressed in eukaryotic cells where it reduces hydrogen and lipid peroxides to alcohols. Adenosine, which is released from stressed or injured cells, protects against ischemia/reperfusion injury and apoptosis. In this study, we hypothesize that the cytoprotective effect of adenosine involves an increase in the activity of GPx-1. Treatment of human primary pulmonary artery endothelial cells (HPAECs) with 50 micromol/L adenosine in the presence of 10 micromol/L erytho-9-(2-hydroxy-3-nonyl)adenine (EHNA), an adenosine deaminase inhibitor, for 48 hours increased GPx-1 mRNA levels 2-fold. GPx-1 protein and enzyme activity also increased approximately 2-fold after treatment. The induction of GPx-1 expression was found to be a consequence of increased mRNA stability and not an increase in transcription. Bisindolylmaleimide I (BIM), a protein kinase C signaling pathway inhibitor, significantly attenuated the induction of GPx-1 mRNA by approximately 36%. The adenosine/EHNA-treated cells were more resistant to hydrogen peroxide stress. Both pharmacological inhibition and siRNA knockdown of GPx-1 attenuated the protective affect of adenosine/EHNA treatment, indicating that the adenosine-induced increase in GPx-1 contributes to an increase in cellular protection against oxidative stress. These data suggest that adenosine may protect the cardiovascular system from ischemia/reperfusion injury, in part, by enhancing the expression of the central intracellular antioxidant enzyme, GPx 1. PMID- 15802614 TI - Immunomodulator FTY720 Induces eNOS-dependent arterial vasodilatation via the lysophospholipid receptor S1P3. AB - The novel immunomodulator FTY720 is effective in experimental models of transplantation and autoimmunity, and is currently undergoing Phase III clinical trials for prevention of kidney graft rejection. FTY720 is a structural analogue of sphingosine-1-phosphate (S1P) and activates several of the S1P receptors. We show that FTY720 induces endothelium-dependent arterial vasodilation in phenylephrine precontracted mouse aortae. Vasodilation did not occur in thoracic aortic rings from eNOS-deficient mice, implicating and effect dependent of activation of the eNOS/NO pathway. Accordingly, FTY720 induced NO release, Akt dependent eNOS phosphorylation and activation in human endothelial cells. For biological efficacy, FTY720 required endogenous phosphorylation, since addition of the sphingosine kinase antagonist N',N-dimethylsphingosine (DMS) prevented activation of eNOS in vitro and inhibited vasodilation in isolated arteries. The endothelial phosphorylation of FTY720 was extremely rapid with almost complete conversion after 10 minutes as determined by mass spectrometry. Finally, we identified the lysophospholipid receptor S1P3 as the S1P receptor responsible for arterial vasodilation by FTY720, as the effect was completely abolished in arteries from S1P3-deficient mice. In summary, we have identified FTY720 as the first immunomodulator for prevention of organ graft rejection in clinical development that, in addition, positively affects the endothelium by stimulating NO production, and thus potentially displaying beneficial effects on transplant survival beyond classical T cell immunosuppression. PMID- 15802615 TI - Regulation of arterial tone by KV1 potassium channels. PMID- 15802616 TI - Candidate gene discovery in cardiovascular disease. PMID- 15802617 TI - Triadin: the new player on excitation-contraction coupling block. PMID- 15802618 TI - Angiotensin II: a devious activator of mineralocorticoid receptor-dependent gene expression. PMID- 15802619 TI - Platelets and chemokines in atherosclerosis: partners in crime. AB - It becomes increasingly evident that blood platelets do not only exert important functions in hemostasis and thrombus formation but are also involved in atherosclerotic vascular disease. A major portion of the underlying mechanisms is related to an intricate functional interaction of platelets with chemokines, which have also been implicated in atherogenesis and neointima formation: (1) Platelets can induce the secretion of chemokines in different cells of the vascular wall; (2) In combination with primary agonists, certain chemokines can potentiate platelet aggregation and adhesion; (3) Activated platelets can release and deposit chemokines and precursors on vascular cell surfaces, which trigger atherogenic recruitment of vascular cells or modulate crucial processes such as angiogenesis and lipoprotein metabolism; (4) Surface-adherent platelets can bind and present vascular cell-derived chemokines to trigger arrest of circulating mononuclear cells. The close linkage between platelets and chemokines as culprits in the pathogenesis of vascular diseases may provide a valuable target for selective interventions. PMID- 15802620 TI - Mechanisms of reactive oxygen species-dependent downregulation of insulin receptor substrate-1 by angiotensin II. AB - OBJECTIVE: Angiotensin II has been implicated in the pathogenesis of the vascular complications of insulin resistance. Recently, serine phosphorylation and degradation of insulin receptor substrate-1 (IRS-1) were shown to inhibit Akt activation and reduce glucose uptake. Therefore, we examined the effects of chronic angiotensin II treatment on IRS-1 phosphorylation and protein expression in vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: Using Western analysis, we found that angiotensin II (100 nmol/L; 18 hours) caused a 61+/-5% degradation of IRS-1 and abolished insulin-induced activation of Akt. Phosphorylation of IRS-1 on Ser307, which leads to subsequent IRS-1 degradation, was stimulated by angiotensin II. This phosphorylation was blocked by the Src inhibitor PP1 and by the antioxidants N-acetylcysteine and ebselen. Stable overexpression of catalase abrogated angiotensin II-induced IRS-1 phosphorylation and IRS-1 degradation. Similarly, a mutant phosphoinositide-dependent kinase-1 (PDK1) that cannot associate with Src abolished IRS-1 phosphorylation and degradation induced by angiotensin II. Proteasome inhibitors also prevented IRS-1 degradation. CONCLUSIONS: Thus, angiotensin II decreases IRS-1 protein levels in VSMCs via Src, PDK1, and reactive oxygen species-mediated phosphorylation of IRS 1 on Ser307 and subsequent proteasome-dependent degradation. These events impair insulin signaling and provide a molecular basis for understanding the clinical observation that angiotensin II type 1 receptor antagonists improve insulin resistance and its associated vasculopathies. PMID- 15802621 TI - Regulation of endothelial cell and endothelial progenitor cell survival and vasculogenesis by integrin-linked kinase. AB - OBJECTIVE: New vessel formation is a dynamic process of attachment, detachment, and reattachment of endothelial cells (ECs) and endothelial progenitor cells (EPCs) with each other and with the extracellular matrix (ECM). Integrin-linked kinase (ILK) plays a pivotal role in ECM-mediated signaling. Therefore, we investigated the role of ILK in ECs and EPCs during neovascularization. METHODS AND RESULTS: In human umbilical cord vein ECs and EPCs, endogenous ILK expression, along with subsequent cell survival signals phospho-Akt and phospho glycogen synthase kinase 3beta, was reduced after anchorage or nutrient deprivation. Even brief anchorage deprivation resulted in retarded capillary tube formation by ECs. Adenoviral ILK gene transfer in ECs and EPCs reversed the decrease in cell survival signals after anchorage or nutrient deprivation, leading to enhanced survival, reduced apoptosis, and significantly accelerated the functional recovery after reattachment. And ILK overexpressing EPCs significantly improved blood flow recovery and prevented limb loss in nude mice hindlimb ischemia model. Furthermore, the efficacy of systemic delivery was equivalent to local injection of ILK-EPCs. CONCLUSIONS: ILK overexpression protects ECs and EPCs from anchorage- or nutrient-deprived stress and enhances neovascularization, suggesting that ILK is an optimal target gene for genetically modified cell-based therapy. Neovascularization is a dynamic process of detachment and reattachment of ECs and EPCs. Endogenous ILK expression was decreased in various stress conditions, and the gene transfer of ILK protected ECs and EPCs from temporary anchorage or nutrient deprivation. Furthermore, ILK gene transfer in EPCs significantly enhanced neovascularization in vivo. PMID- 15802622 TI - Extracellular proteases in atherosclerosis and restenosis. AB - Extracellular proteolysis plays a key role in many pathophysiologic processes including cancer, inflammatory diseases, and cardiovascular conditions such as atherosclerosis and restenosis. Whereas matrix metalloproteinases are their best known member, many others are becoming better known. The extracellular proteases are a complex and heterogeneous superfamily of enzymes. They include metalloproteinases (matrix metalloproteinases, adamalysins, or pappalysins), serine proteases (elastase, coagulation factors, plasmin, tissue plasminogen activator, urokinase plasminogen activator), and the cysteine proteases (such cathepsins). In addition to their matrix degradation capabilities, they have other less well known biologic functions that include angiogenesis, growth factor bioavailability, cytokine modulation, receptor shedding, enhancing cell migration, proliferation, invasion, and apoptosis. This review discusses extracellular proteases relevant to the vasculature, their classification and function, and how protease disorders contribute to arterial plaque growth, including chronic atherosclerosis, acute coronary syndromes, restenosis, and vascular remodeling. These broad extracellular protease functions make them potentially interesting therapeutic targets. PMID- 15802623 TI - Autocrine and paracrine transcriptional regulation of type IIA secretory phospholipase A2 gene in vascular smooth muscle cells. AB - OBJECTIVE: The inflammation that occurs during the development of atherosclerosis is characterized by a massive release of sPLA2-IIA (group IIA secretory phospholipase A2) from vascular smooth muscle cells (VSMCs). We have investigated the autocrine function of sPLA2-IIA in rat aortic and human VSMCs. METHODS AND RESULTS: We found that the transcription of the endogenous sPLA2-IIA gene increased by adding a cell supernatant containing human sPLA2-IIA proteins. We show that this effect was independent of the sPLA2 activity using sPLA2-IIA proteins lacking enzyme activity. Transient transfections with various sPLA2-IIA rat promoter-luciferase constructs demonstrated that the C/EBP, NK-kappaB, and Ets transcription factors are involved in the increase in sPLA2-IIA gene transcription. We also found the M-type sPLA2 receptor mRNA in VSMCs, and we showed that the sPLA2-luciferase reporter gene was induced by the specific agonist of the sPLA2 receptor, aminophenylmannopyranoside (APMP), and that this induction was mediated by the same transcription factor-binding sites. Finally, we used a sPLA2-IIA mutant unable to bind heparan-sulfate proteoglycans to show that the binding of wild-type sPLA2-IIA to proteoglycans is essential for the induction of an autocrine loop. CONCLUSIONS: We have thus identified new autocrine and paracrine pathways activating sPLA2-IIA gene expression in rat and human VSMCs. PMID- 15802624 TI - Elevated serum C-reactive protein levels and advanced atherosclerosis in youth. AB - OBJECTIVE: To determine the associations among serum C-reactive protein (CRP) concentration, age, sex, risk factors for coronary heart disease (CHD), and atherosclerosis in young people. METHODS AND RESULTS: In 1244 subjects 15 to 34 years of age, we measured gross atherosclerotic lesions in the right coronary artery (RCA) and abdominal aorta (AA) and American Heart Association (AHA) lesion grade in the left anterior descending (LAD) coronary artery; serum CRP, lipoprotein cholesterol, and thiocyanate (for smoking) concentrations; intimal thickness of renal arteries (for hypertension); glycohemoglobin (for hyperglycemia); and body mass index (for obesity). Serum CRP levels increased with age, were higher in women than in men, and were positively related to obesity and hyperglycemia. Serum CRP > or =10 mg/L was associated with more extensive gross raised lesions in the RCA after age 25 and in the AA after age 30. Serum CRP > or =3 was associated with a greater prevalence of AHA grade 5 lesions in the proximal LAD coronary artery after age 25. The associations of CRP with lesions were independent of the traditional CHD risk factors. CONCLUSIONS: Serum CRP level is independently associated with advanced atherosclerosis in young persons. PMID- 15802625 TI - A new cellular signaling mechanism for angiotensin II activation of NF-kappaB: An IkappaB-independent, RSK-mediated phosphorylation of p65. AB - OBJECTIVE: Angiotensin II (Ang II) promotes vascular inflammation and remodeling via activation of nuclear factor kappaB (NF-kappaB)-mediated transcription of proinflammatory genes such as interleukin-6 (IL-6). We examined the signaling mechanism whereby Ang II activates NF-kappaB in vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: Ang II treatment did not increase phosphorylation of inhibitor of kappaBalpha (IkappaBalpha) or IkappaBbeta or decrease their levels. In contrast, mitogen-activated protein kinase kinase-1 (MEK1) inhibition (dominant-negative MEK1 adenovirus or inhibitor U0126) suppressed Ang II-induced NF-kappaB promoter activity, NF-kappaB DNA-binding activity, p65 phosphorylation, and led to 70% reduction in IL-6 transcription/production. The mechanism involved Ang II activation of Ras and MEK1. Signaling distal to MEK1 involved extracellular signal-regulated kinase (ERK) because inhibition of MEK1 suppressed the Ang II-induced activation of ribosomal S6 kinase (RSK), a substrate of ERK. Downregulation of RSK by small interfering RNA (SiRNA) in VSMCs was found to suppress Ang II-induced activation of NF-kappaB and p65 phosphorylation. Immunopurified RSK from Ang II-treated VSMCs phosphorylated recombinant glutathione S-transferase-p65 in vitro. CONCLUSIONS: We uncovered a nonclassical signaling pathway (Ras/MEK1/ERK/RSK) from Ang II to activation of NF-kappaB, a mechanism by which Ang II stimulates RSK-mediated phosphorylation of p65 to participate in vascular inflammation. PMID- 15802626 TI - C-reactive protein-induced in vitro endothelial cell activation is an artefact caused by azide and lipopolysaccharide. AB - OBJECTIVE: C-reactive protein (CRP) has been proposed to be an independent risk factor for cardiovascular disease. In vitro studies investigating the mechanism behind this have used purified commercial CRP (cCRP) and endothelial cells. We investigated the role of contaminants in cCRP preparations. METHODS AND RESULTS: Human umbilical vein endothelial cells and the human endothelial cell line EA.hy926 were incubated with Escherichia coli-derived cCRP, in-house-generated azide-free recombinant, and ascites-purified CRP, azide, or lipopolysaccharide (LPS) equivalent to the concentration present in cCRP preparations. Cells were investigated for change in cell proliferation, morphology, apoptosis, and expression of endothelial NO synthase and intercellular adhesion molecule-1. Cell supernatants were assessed for monocyte chemoattractant protein-1 (MCP-1), interleukin-8, von Willebrand factor secretion, and pH change. Only cCRP was able to induce all activation events analyzed; however, this ability was lost on extensive dialysis, suggesting that low molecular weight contaminants were responsible for these events. Indeed, the effects of cCRP were mirrored by azide or LPS. CONCLUSIONS: We investigated a wide range of effects on endothelial cells ascribed to CRP; however, azide and LPS, but never CRP itself, were responsible for the cell activation events. We conclude that CRP, per se, does not activate endothelial cells. PMID- 15802627 TI - An ultrasensitive site-specific DNA recombination assay based on dual-color fluorescence cross-correlation spectroscopy. AB - Site-specific exchange of genetic information is mediated by DNA recombinases, such as FLP or Cre, and has become a valuable tool in modern molecular biology. The so far low number of suitable recombinating enzymes has driven current research activities towards alteration of catalytic properties, such as thermostability or recognition sequences. However, identification and analysis of new mutants requires sensitive in vitro activity assays, which traditionally are based on gel electrophoresis. Here, we describe the development of a new sensitive DNA recombination assay based on dual-color fluorescence cross correlation spectroscopy (DC-FCCS), which works in homogenous solution and does not require any separation step such as electrophoresis. The assay was validated with unlabeled FLP recombinase and different fluorescently labeled DNA substrates containing the FLP recognition target (FRT). This strategy fulfills all requirements for possible application in high throughput screening and engineering of new site-specific DNA recombinases starting from the FLP-FRT system, and is easily adjustable to other systems like Cre/loxP. PMID- 15802631 TI - Natriuretic peptides and mortality after stroke. AB - BACKGROUND AND PURPOSE: Measurement of natriuretic peptides provides prognostic information in various patient populations. The prognostic value of natriuretic peptides among patients with acute stroke is not known, although elevated peptide levels have been observed. METHODS: A series of 51 patients (mean age, 68+/-11 years) with first-ever ischemic stroke underwent a comprehensive clinical examination and measurements of plasma atrial natriuretic peptides (N-ANP) and brain natriuretic peptides (N-BNP) in the acute phase of stroke. The patients were followed-up for 44+/-21 months. Risk factors for all-cause mortality were assessed. Control populations, matched for gender and age, consisted of 51 patients with acute myocardial infarction (AMI) and 25 healthy subjects. RESULTS: Plasma concentrations of N-ANP (mean+/-SD, 988+/-993 pmol/L) and N-BNP (751+/ 1608 pmol/L) in the stroke patients were at the same level as those in the AMI patients (NS for both), but significantly higher than those of the healthy subjects (358+/-103 pmol/L, P<0.001 and 54+/-26 pmol/L, P<0.01, respectively). Elevated levels of N-ANP and N-BNP predicted mortality after stroke (risk ratio [RR] 4.3, P<0.01 and RR 3.9, P<0.01, respectively) and after AMI (P<0.05), and remained independent predictors of death after stroke even after adjustment for age, diabetes, coronary artery disease, and medication (RR 3.9, P<0.05 and RR 3.7, P<0.05, respectively). CONCLUSIONS: Plasma levels of natriuretic peptides are elevated in the acute phase of stroke and predict poststroke mortality. PMID- 15802632 TI - Phosphodiesterase 4D gene, ischemic stroke, and asymptomatic carotid atherosclerosis. AB - BACKGROUND AND PURPOSE: Phosphodiesterase 4D (PDE4D) was identified recently as the first novel stroke gene to predispose to ischemic stroke independently of conventional risk factors. An association was only found with large vessel and cardioembolic stroke, suggesting a mechanism of accelerated atherosclerosis. We sought to replicate this association in ischemic stroke as a whole, and individual stroke subtypes, in a non-Icelandic European population. To assess a role in early atherosclerosis, we also sought associations with underlying asymptomatic atherosclerosis itself, assessed by carotid ultrasound in a community population. METHODS: A total of 737 consecutive white patients with stroke and 933 white community controls free of symptomatic cerebrovascular disease were examined using a case control methodology. For association with atherosclerosis, intima-media thickness (IMT) in a community population (n=1000) was assessed using carotid ultrasound. Nineteen single nucleotide polymorphisms (SNPs) and 1 minisatellite in the PDE4D gene were determined, with haplotyping undertaken using Phase 2.0. RESULTS: No association with ischemic stroke overall was identified. Six of the 19 SNPs were associated with cardioembolic stroke and 2 different SNPs with large vessel disease. There was no association with carotid artery IMT or carotid plaque in the asymptomatic community subjects. CONCLUSIONS: The PDE4D gene is not a major risk factor for ischemic stroke, or early atherosclerosis, within the 2 European population samples studied. On analysis of individual stroke subtypes, there is a possible association with cardioembolic stroke, but the lack of association with carotid IMT and plaque would suggest that this is via a mechanism other than accelerated atherosclerosis. PMID- 15802633 TI - Assessment of dural arteriovenous fistulae by transcranial color-coded duplex sonography. AB - BACKGROUND AND PURPOSE: To study hemodynamic changes and to determine the value of contrast-enhanced transcranial color-coded sonography (TCCS) for the evaluation of dural arteriovenous fistulae (DAVF) before and after transcatheter embolization. METHODS: Twenty-four patients (mean age 61+/-11 years) with occipitally located DAVF were studied with contrast-enhanced TCCS using the transtemporal bone window in transverse-axial and coronal insonation planes. Blood flow velocity measurements of all depictable basal cerebral veins and sinuses were obtained before and after transcatheter embolization. Pretreatment and post-treatment flow velocity values were compared. RESULTS: Four of the 24 patients (17%) could not be studied because of an insufficient temporal bone window. In all remaining patients (n=20), draining veins/sinuses could be identified because of pathologically increased blood flow velocities with peak systolic flow velocities of >50 cm/s. Of the 27 draining vessels depicted by DSA, TCCS correctly identified 25 (93%): the basal vein (3 of 3), the straight sinus (3 of 3), the superior sagittal sinus (1 of 3), the transverse sinus (9), the sigmoid sinus (4), and the superior petrosal sinus (5/5). However, TCCS failed to depict supplementary drainage via cortical veins. After transcatheter embolization, mean reduction of blood flow velocity was 44+/-18% (P<0.01) compared with pretreatment values. CONCLUSIONS: Contrast-enhanced TCCS is a promising technique for monitoring embolization of DAVF, follow-up after complete fistula occlusion, and may even be useful as a screening tool in patients with pulsatile tinnitus. PMID- 15802634 TI - Differential diagnosis of stroke in a setting of high HIV prevalence in Blantyre, Malawi. AB - BACKGROUND AND PURPOSE: The differential diagnosis of stroke in Africa in areas with high HIV prevalence includes brain infections. We studied causes of stroke in Blantyre, Malawi, where HIV prevalence among medical inpatients is 70%. METHODS: In a descriptive study of 8-month duration, all patients presenting at Queen Elizabeth Central Hospital, Blantyre, with central neurological deficit of acute onset (<24 hours) had baseline investigations, including full blood count, blood glucose, serology for toxoplasmosis, syphilis, and HIV, ECG, echocardiogram, ultrasound of the carotid arteries, and computerized tomography scan of the brain. A lumbar puncture was performed unless contraindicated. RESULTS: Ninety-eight consecutive patients (49 males) were studied. In those who were HIV positive (48%), the mean age was 37.5 years; ischemic stroke was the commonest diagnosis (n=25; 58%), followed by infection (n=11; 23%; including tuberculous [n=4] and cryptococcal [n=2] meningitis; toxoplasmic encephalitis [n=1]; neurocysticercosis [n=1]; brain abscess [n=1]; and progressive multifocal leucoencephalopathy [n=2]). No clinical or laboratory parameters could be identified as predictors for infection, but 3 of 5 patients with fever on admission had tuberculous meningitis. In HIV-negative patients (mean age 58.6 years), 55% had ischemic stroke and 31% had intracerebral hemorrhage; no brain infection was diagnosed. Presence of vascular disease correlated with age but not with HIV status. CONCLUSIONS: Ischemic stroke was found in half of patients irrespective of HIV status. In those who are HIV positive, brain infection should be considered for which the presence of fever and examination of cerebrospinal fluid seem most useful in diagnosis. PMID- 15802635 TI - Body fat distribution and long-term risk of stroke mortality. AB - BACKGROUND AND PURPOSE: Excess weight is an important determinant of cardiovascular disease, but the relationship between excess weight, its distribution, and stroke is yet unclear. We examined in a large prospective cohort study the association between body fat distribution and stroke mortality among middle-aged men. METHODS: A cohort of male civil servants and municipal employees free of cardiovascular disease in Israel (n=9151) were followed up for mortality over 23 years. The subscapular skinfold (SSF) was used as a measure of trunk and overall obesity and the ratio of subscapular to triceps skinfold thickness (SFR) as an indicator of trunk versus peripheral distribution of body fat. RESULTS: During the follow-up period, 316 died of stroke, and 865 died of coronary heart disease. The estimated age-adjusted hazard ratios (HRs) for stroke mortality, associated with 1 SD increment of SSF, was 1.12 (95% CI, 1.01 to 1.25) and for body mass index, 1.17 (1.06 to 1.30), but these associations were markedly weakened when adjusting for blood pressure. SFR was associated with an age-adjusted HR for stroke mortality of 1.14 (1.03 to 1.26). Further adjusting for systolic blood pressure, diabetes mellitus, cigarette smoking, and socioeconomic status (HR, 1.11; 1.01 to 1.23) as well as body mass index (HR, 1.11; 1.00 to 1.23) only mildly attenuated this association. Subjects with SFR in the upper quartile exhibited a approximately 1.5-fold higher adjusted HR (1.53; 1.10 to 2.12) compared with the lowest quartile. CONCLUSIONS: Indices of body fat and body fat distribution predict long-term stroke and coronary heart disease mortality among middle-aged men. SFR, an indicator of trunk versus peripheral distribution of body fat, is associated with stroke mortality, independent of main mediators of the effect of obesity on health and of body mass index. PMID- 15802636 TI - Intra-arterial thrombolytic therapy in peri-coronary angiography ischemic stroke. AB - BACKGROUND: Intra-arterial thrombolysis (IAT) for peri-coronary angiography (CA) stroke may be safe and efficacious. However, IAT may increase the risk of intracranial hemorrhage (ICH). METHODS: A retrospective study was performed involving 3 university hospitals. All peri-CA IAT-treated cases were identified. Patient demographics, stroke severity, angiographic findings, thrombolytic use, modified Rankin Scale (mRS), ICH, and mortality were determined. RESULTS: A total of 21 patients with post-left CA stroke were treated with IAT (mean age 71.8+/ 12.3 years). Arterial occlusion was found in 14 (66.7%) and 7 (33.3%) of the anterior and posterior circulation, respectively. Mean time-to-therapy was 36+/ 12 minutes from the time the neurological deficit was noted. mRS < or =2 occurred in 10 of 21 (48%) patients. Patients with younger age and shorter time-to-IAT had more complete arterial recanalization and clinical recovery. Symptomatic ICH occurred in 3 (14%) cases, and 4 (19%) patients died. CONCLUSIONS: Peri-CA IAT appears to be feasible and safe without increased risk of symptomatic ICH and death when compared with the previously reported IAT literature. PMID- 15802637 TI - Management of depression after stroke: a systematic review of pharmacological therapies. AB - BACKGROUND AND PURPOSE: Although depression may affect recovery and outcome after stroke, it is often overlooked or inadequately managed, and there is uncertainty regarding the benefits of antidepressant therapy in this setting. We aimed to assess the effectiveness of antidepressants for the treatment and prevention of depression after stroke. METHODS: We undertook a systematic review using Cochrane methods of randomized placebo-controlled trials of antidepressants for the treatment or prevention of depressive illness and "abnormal mood" after stroke. Treatment effects on physical and other outcomes were also examined. RESULTS: Outcome data were available for 7 treatment trials including 615 patients and 9 prevention trials including 479 patients. Because of the considerable variation in research design, trial quality, and method of reporting across studies, we did not pool all the outcome data. In the treatment trials, antidepressants reduced mood symptoms but had no clear effect on producing a remission of diagnosable depressive illness. There was no definitive evidence that antidepressants prevent depression or improve recovery after stroke. CONCLUSIONS: There is insufficient randomized evidence to support the routine use of antidepressants for the prevention of depression or to improve recovery from stroke. Although antidepressants may improve mood in stroke patients with depression, it is unclear how clinically significant such modest effects are in patients other than those with major depression. There is a pressing need for further research to better define the role of antidepressants in stroke management. PMID- 15802638 TI - Using mismatch on MRI to select thrombolytic responders: an attractive hypothesis awaiting confirmation. PMID- 15802639 TI - Autonomic function is impaired in elderly stroke survivors. AB - BACKGROUND AND PURPOSE: Impaired autonomic function is common in the acute poststroke phase but little is known about the longer term effects, particularly in older people. We sought to determine if autonomic function is impaired after stroke recovery in older patients. METHODS: A cross-sectional case-control study comparing autonomic function in 76 nondemented stroke patients with 70 community living controls aged older than 75 years. RESULTS: Cases were assessed on average 9 months after stroke. From power spectral analysis of heart rate variability, stroke patients had lower total (P=0.032) and low-frequency (P=0.014) spectral densities and impaired baroreflex sensitivity (alpha low-frequency baroreflex sensitivity, P=0.006). From a series of cardiovascular autonomic reflex tests, heart rate variation during forced respiration, Valsalva ratio, and blood pressure overshoot during Valsalva maneuver were significantly lower in stroke patients (P=0.003, <0.001, and 0.027, respectively). Blood pressure response to isometric exercise was significantly exaggerated in stroke patients (P=0.007). CONCLUSIONS: Cardiovascular autonomic function is impaired long after the index event in stroke survivors. Impaired autonomic function may increase the risk of all-cause mortality and cardiovascular mortality in older stroke survivors. PMID- 15802640 TI - Identification of mouse brain proteins associated with isoform 3 of metallothionein. AB - Using immunological approaches and mass spectrometry, five proteins associated with metallothionein-3 in mouse brains have been identified. Metallothionein-3 and associated proteins were isolated using immunoaffinity chromatography over immobilized anti-mouse brain MT3 antibody. Proteins in the recovered pool were separated by SDS-polyacrylamide gel electrophoresis, and distinct bands were excised and the proteins digested using trypsin. Peptides were extracted and analyzed using electrospray ionization mass spectrometry. Initial identification was done comparing the identified peptide mass:charge ratios to the MASCOT database. Confirmation of proteins was accomplished by sequencing of selected peptides using tandem mass spectrometry and comparison to the MASCOT database. The proteins were heat-shock protein 84 (mouse variant of heat-shock protein 90), heat-shock protein 70, dihydropyrimidinase-like protein 2, creatine kinase, and beta actin. Independently using antibodies against metallothionein-3, creatine kinase, and heat-shock protein 84 showed that all three proteins were coimmunoprecipitated from whole mouse brain homogenates with each of the three antibodies. Mixing purified samples of metallothionein and human brain creatine kinase also generated a complex that could be immunoprecipitated either by anti metallothionein-3 or anticreatine kinase antibody. These data are consistent with metallothionein-3 being present in the mouse brain as part of a multiprotein complex providing new functional information for understanding the role of metallothionein-3 in neuronal physiology. PMID- 15802642 TI - Multiple equilibria of the Escherichia coli chaperonin GroES revealed by mass spectrometry. AB - Nanospray time-of-flight mass spectrometry has been used to study the assembly of the heptamer of the Escherichia coli cochaperonin protein GroES, a system previously described as a monomer-heptamer equilibrium. In addition to the monomers and heptamers, we have found measurable amounts of dimers and hexamers, the presence of which suggests the following mechanism for heptamer assembly: 2 Monomers <--> Dimer; 3 Dimers <--> Hexamer; Hexamer + Monomer <--> Heptamer. Equilibrium constants for each of these steps, and an overall constant for the Monomer <--> Heptamer equilibrium, have been estimated from the data. These constants imply a standard free-energy change, DeltaG(0), of about 9 kcal/mol for each contact surface formed between GroES subunits, except for the addition of the last subunit, where DeltaG(0) = 6 kcal/mol. This lower value probably reflects the loss of entropy when the heptamer ring is formed. These experiments illustrate the advantages of electrospray mass spectrometry as a method of measuring all components of a multiple equilibrium system. PMID- 15802641 TI - Compensating increases in protein backbone flexibility occur when the Dead ringer AT-rich interaction domain (ARID) binds DNA: a nitrogen-15 relaxation study. AB - AT-rich interaction domains (ARIDs) are found in a large number of eukaryotic transcription factors that regulate cell proliferation, differentiation, and development. Previously we elucidated how ARIDs recognize DNA by determining the solution structure of the Drosophila melanogaster Dead ringer protein in both its DNA-free and -bound states. In order to quantitatively determine how ARIDs alter their mobility to recognize DNA, we have measured the relaxation parameters of the backbone nitrogen-15 nuclei of Dead ringer in its free and bound forms, and interpreted these data using the model-free approach. We show that Dead ringer undergoes significant changes in its mobility upon binding, with residues in the loop connecting helices H5 and H6 becoming immobilized in the major groove and contacts to the minor groove slowing down the motion of residues at the C terminus. A DNA-induced rotation and displacement of the N-terminal subdomain of the protein increases the mobility of helix H1 located distal to the DNA interface and may partially negate the entropic cost of immobilizing interfacial residues. Elevated motions on the micro- to millisecond timescale in the N terminal domain prior to DNA binding appear to foreshadow the DNA-induced conformation change. PMID- 15802643 TI - The crystal structure of a partial mouse Notch-1 ankyrin domain: repeats 4 through 7 preserve an ankyrin fold. AB - Folding and stability of proteins containing ankyrin repeats (ARs) is of great interest because they mediate numerous protein-protein interactions involved in a wide range of regulatory cellular processes. Notch, an ankyrin domain containing protein, signals by converting a transcriptional repression complex into an activation complex. The Notch ANK domain is essential for Notch function and contains seven ARs. Here, we present the 2.2 A crystal structure of ARs 4-7 from mouse Notch 1 (m1ANK). These C-terminal repeats were resistant to degradation during crystallization, and their secondary and tertiary structures are maintained in the absence of repeats 1-3. The crystallized fragment adopts a typical ankyrin fold including the poorly conserved seventh AR, as seen in the Drosophila Notch ANK domain (dANK). The structural preservation and stability of the C-terminal repeats shed a new light onto the mechanism of hetero-oligomeric assembly during Notch-mediated transcriptional activation. PMID- 15802644 TI - Synthetic prions generated in vitro are similar to a newly identified subpopulation of PrPSc from sporadic Creutzfeldt-Jakob Disease. AB - In recent studies, the amyloid form of recombinant prion protein (PrP) encompassing residues 89-230 (rPrP 89-230) produced in vitro induced transmissible prion disease in mice. These studies showed that unlike "classical" PrP(Sc) produced in vivo, the amyloid fibrils generated in vitro were more proteinase-K sensitive. Here we demonstrate that the amyloid form contains a proteinase K-resistant core composed only of residues 152/153-230 and 162-230. The PK-resistant fragments of the amyloid form are similar to those observed upon PK digestion of a minor subpopulation of PrP(Sc) recently identified in patients with sporadic Creutzfeldt-Jakob disease (CJD). Remarkably, this core is sufficient for self-propagating activity in vitro and preserves a beta-sheet-rich fibrillar structure. Full-length recombinant PrP 23-230, however, generates two subpopulations of amyloid in vitro: One is similar to the minor subpopulation of PrP(Sc), and the other to classical PrP(Sc). Since no cellular factors or templates were used for generation of the amyloid fibrils in vitro, we speculate that formation of the subpopulation of PrP(Sc) with a short PK-resistant C terminal region reflects an intrinsic property of PrP rather than the influence of cellular environments and/or cofactors. Our work significantly increases our understanding of the biochemical nature of prion infectious agents and provides a fundamental insight into the mechanisms of prions biogenesis. PMID- 15802645 TI - Unfolding of Green Fluorescent Protein mut2 in wet nanoporous silica gels. AB - Many of the effects exerted on protein structure, stability, and dynamics by molecular crowding and confinement in the cellular environment can be mimicked by encapsulation in polymeric matrices. We have compared the stability and unfolding kinetics of a highly fluorescent mutant of Green Fluorescent Protein, GFPmut2, in solution and in wet, nanoporous silica gels. In the absence of denaturant, encapsulation does not induce any observable change in the circular dichroism and fluorescence emission spectra of GFPmut2. In solution, the unfolding induced by guanidinium chloride is well described by a thermodynamic and kinetic two-state process. In the gel, biphasic unfolding kinetics reveal that at least two alternative conformations of the native protein are significantly populated. The relative rates for the unfolding of each conformer differ by almost two orders of magnitude. The slower rate, once extrapolated to native solvent conditions, superimposes to that of the single unfolding phase observed in solution. Differences in the dependence on denaturant concentration are consistent with restrictions opposed by the gel to possibly expanded transition states and to the conformational entropy of the denatured ensemble. The observed behavior highlights the significance of investigating protein function and stability in different environments to uncover structural and dynamic properties that can escape detection in dilute solution, but might be relevant for proteins in vivo. PMID- 15802646 TI - Enterococcus faecalis phosphomevalonate kinase. AB - The six enzymes of the mevalonate pathway of isopentenyl diphosphate biosynthesis represent potential for addressing a pressing human health concern, the development of antibiotics against resistant strains of the Gram-positive streptococci. We previously characterized the first four of the mevalonate pathway enzymes of Enterococcus faecalis, and here characterize the fifth, phosphomevalonate kinase (E.C. 2.7.4.2). E. faecalis genomic DNA and the polymerase chain reaction were used to clone DNA thought to encode phosphomevalonate kinase into pET28b(+). Double-stranded DNA sequencing verified the sequence of the recombinant gene. The encoded N-terminal hexahistidine-tagged protein was expressed in Escherichia coli with induction by isopropylthiogalactoside and purified by Ni(++) affinity chromatography, yield 20 mg protein per liter. Analysis of the purified protein by MALDI-TOF mass spectrometry established it as E. faecalis phosphomevalonate kinase. Analytical ultracentrifugation revealed that the kinase exists in solution primarily as a dimer. Assay for phosphomevalonate kinase activity used pyruvate kinase and lactate dehydrogenase to couple the formation of ADP to the oxidation of NADH. Optimal activity occurred at pH 8.0 and at 37 degrees C. The activation energy was approximately 5.6 kcal/mol. Activity with Mn(++), the preferred cation, was optimal at about 4 mM. Relative rates using different phosphoryl donors were 100 (ATP), 3.6 (GTP), 1.6 (TTP), and 0.4 (CTP). K(m) values were 0.17 mM for ATP and 0.19 mM for (R,S)-5-phosphomevalonate. The specific activity of the purified enzyme was 3.9 micromol substrate converted per minute per milligram protein. Applications to an immobilized enzyme bioreactor and to drug screening and design are discussed. PMID- 15802647 TI - Improved side-chain modeling for protein-protein docking. AB - Success in high-resolution protein-protein docking requires accurate modeling of side-chain conformations at the interface. Most current methods either leave side chains fixed in the conformations observed in the unbound protein structures or allow the side chains to sample a set of discrete rotamer conformations. Here we describe a rapid and efficient method for sampling off-rotamer side-chain conformations by torsion space minimization during protein-protein docking starting from discrete rotamer libraries supplemented with side-chain conformations taken from the unbound structures, and show that the new method improves side-chain modeling and increases the energetic discrimination between good and bad models. Analysis of the distribution of side-chain interaction energies within and between the two protein partners shows that the new method leads to more native-like distributions of interaction energies and that the neglect of side-chain entropy produces a small but measurable increase in the number of residues whose interaction energy cannot compensate for the entropic cost of side-chain freezing at the interface. The power of the method is highlighted by a number of predictions of unprecedented accuracy in the recent CAPRI (Critical Assessment of PRedicted Interactions) blind test of protein protein docking methods. PMID- 15802648 TI - The P5 protein from bacteriophage phi-6 is a distant homolog of lytic transglycosylases. AB - Peptidases are classical objects of enzymology and structural studies. However, a few protein families with experimentally characterized proteolytic activity, but unknown catalytic mechanism and three-dimensional structures, still exist. Using comparative sequence analysis, we deduce spatial structure for one of such families, namely, U40, which contains just one P5 protein from bacteriophage phi 6. We show that this singleton sequence possesses conserved sequence motifs characteristic of lysozymes and is a distant homolog of lytic transglycosylases that cleave bacterial peptidoglycan. The structure of the P5 protein is therefore predicted to adopt the lysozyme-like fold shared by T4, lambda, C-type, G-type lysozymes, and lytic transglycosylases. Since previous biochemical experiments with P5 of phi-6 have indicated that the purified enzyme possesses endopeptidase activity and not glycosidase activity, our results point to the possibility of a newly evolved molecular function and call for further experimental characterization of this unusual P5 protein. PMID- 15802649 TI - One- and two-body decomposable Poisson-Boltzmann methods for protein design calculations. AB - Successfully modeling electrostatic interactions is one of the key factors required for the computational design of proteins with desired physical, chemical, and biological properties. In this paper, we present formulations of the finite difference Poisson-Boltzmann (FDPB) model that are pairwise decomposable by side chain. These methods use reduced representations of the protein structure based on the backbone and one or two side chains in order to approximate the dielectric environment in and around the protein. For the desolvation of polar side chains, the two-body model has a 0.64 kcal/mol RMSD compared to FDPB calculations performed using the full representation of the protein structure. Screened Coulombic interaction energies between side chains are approximated with an RMSD of 0.13 kcal/mol. The methods presented here are compatible with the computational demands of protein design calculations and produce energies that are very similar to the results of traditional FDPB calculations. PMID- 15802650 TI - Action-at-a-distance interactions enhance protein binding affinity. AB - The identification of protein mutations that enhance binding affinity may be achieved by computational or experimental means, or by a combination of the two. Sources of affinity enhancement may include improvements to the net balance of binding interactions of residues forming intermolecular contacts at the binding interface, such as packing and hydrogen-bonding interactions. Here we identify noncontacting residues that make substantial contributions to binding affinity and that also provide opportunities for mutations that increase binding affinity of the TEM1 beta-lactamase (TEM1) to the beta-lactamase inhibitor protein (BLIP). A region of BLIP not on the direct TEM1-binding surface was identified for which changes in net charge result in particularly large increases in computed binding affinity. Some mutations to the region have previously been characterized, and our results are in good correspondence with this results of that study. In addition, we propose novel mutations to BLIP that were computed to improve binding significantly without contacting TEM1 directly. This class of noncontacting electrostatic interactions could have general utility in the design and tuning of binding interactions. PMID- 15802651 TI - Visualization of conformational distribution of short to medium size segments in globular proteins and identification of local structural motifs. AB - Analysis of the conformational distribution of polypeptide segments in a conformational space is the first step for understanding a principle of structural diversity of proteins. Here, we present a statistical analysis of protein local structures based on interatomic C(alpha) distances. Using principal component analysis (PCA) on the intrasegment C(alpha)-C(alpha) atomic distances, the conformational space of protein segments, which we call the protein segment universe, has been visualized, and three essential coordinate axes, suitable for describing the universe, have been identified. Three essential axes specified radius of gyration, structural symmetry, and separation of hairpin structures from other structures. Among the segments of arbitrary length, 6-22 residues long, the conservation of those axes was uncovered. Further application of PCA to the two largest clusters in the universe revealed local structural motifs. Although some of motifs have already been reported, we identified a possibly novel strand motif. We also showed that a capping box, which is one of the helix capping motifs, was separated into independent subclusters based on the C(alpha) geometry. Implications of the strand motif, which may play a role for protein protein interaction, are discussed. The currently proposed method is useful for not only mapping the immense universe of protein structures but also identification of structural motifs. PMID- 15802652 TI - Characterization of the HslU chaperone affinity for HslV protease. AB - The HslVU complex is a bacterial two-component ATP-dependent protease, consisting of HslU chaperone and HslV peptidase. Investigation of protein-protein interactions using SPR in Escherichia coli HslVU and the protein substrates demonstrates that HslU and HslV have moderate affinity (Kd = 1 microM) for each other. However, the affinity of HslU for HslV fivefold increased (Kd approximately 0.2 microM) after binding with the MBP approximately SulA protein indicating the formation of a "ternary complex" of HslV-HslU-MBP approximately SulA. The molecular interaction studies also revealed that HslU strongly binds to MBP approximately SulA with 10(-9) M affinity but does not associate with nonstructured casein. Conversely, HslV does not interact with the MBP-SulA whereas it strongly binds with casein (Kd = 0.2 microM) requiring an intact active site of HslV. These findings provide evidence for "substrate-induced" stable HslVU complex formation. Presumably, the binding of HslU to MBP approximately SulA stimulates a conformational change in HslU to a high-affinity form for HslV. PMID- 15802653 TI - Deletion of the proline-rich region of TonB disrupts formation of a 2:1 complex with FhuA, an outer membrane receptor of Escherichia coli. AB - TonB protein of Escherichia coli couples the electrochemical potential of the cytoplasmic membrane (CM) to active transport of iron-siderophores and vitamin B(12) across the outer membrane (OM). TonB interacts with OM receptors and transduces conformationally stored energy. Energy for transport is provided by the proton motive force through ExbB and ExbD, which form a ternary complex with TonB in the CM. TonB contains three distinct domains: an N-terminal signal/anchor sequence, a C-terminal domain, and a proline-rich region. The proline-rich region was proposed to extend TonB's structure across the periplasm, allowing it to contact spatially distant OM receptors. Having previously identified a 2:1 stoichiometry for the complex of full-length (FL) TonB and the OM receptor FhuA, we now demonstrate that deletion of the proline-rich region of TonB (TonBDelta66 100) prevents formation of the 2:1 complex. Sedimentation velocity analytical ultracentrifugation of TonBDelta66-100 with FhuA revealed that a 1:1 TonB-FhuA complex is formed. Interactions between TonBDelta66-100 and FhuA were assessed by surface plasmon resonance, and their affinities were determined to be similar to those of TonB (FL)-FhuA. Presence of the FhuA-specific siderophore ferricrocin altered neither stoichiometry nor affinity of interaction, leading to our conclusion that the proline-rich region in TonB is important in forming a 2:1 high-affinity TonB-FhuA complex in vitro. Furthermore, TonBDelta66-100 FhuADelta21-128 interactions demonstrated that the cork region of the OM receptor was also important in forming a complex. Together, these results demonstrate a novel function of the proline-rich region of TonB in mediating TonB-TonB interactions within the TonB-FhuA complex. PMID- 15802654 TI - Crystal structure of yeast YHR049W/FSH1, a member of the serine hydrolase family. AB - Yhr049w/FSH1 was recently identified in a combined computational and experimental proteomics analysis for the detection of active serine hydrolases in yeast. This analysis suggested that FSH1 might be a serine-type hydrolase belonging to the broad functional alphabeta-hydrolase superfamily. In order to get insight into the molecular function of this gene, it was targeted in our yeast structural genomics project. The crystal structure of the protein confirms that it contains a Ser/His/Asp catalytic triad that is part of a minimal alpha/beta-hydrolase fold. The architecture of the putative active site and analogies with other protein structures suggest that FSH1 may be an esterase. This finding was further strengthened by the unexpected presence of a compound covalently bound to the catalytic serine in the active site. Apparently, the enzyme was trapped with a reactive compound during the purification process. PMID- 15802655 TI - The effects of social change within a group on membership preferences: to leave or not to leave? AB - Two studies examined immediate (Study 1) and long-term (Study 2) behavioral consequences of previously documented asymmetries in cognitive and evaluative reactions to change in majority and minority positions within a group. Study 1 found an overall decrease in preferences for group membership immediately following change, which was preceded by decategorization and devaluation of the group in response to loss, together with lack of categorization and positive evaluation in response to gain of the majority position. Study 2 found a gradual increase in preference to stay with (vs. exit) the group among former minorities with prolonged interactions that confirmed their gained majority position. A gradual increase in preference for group membership was paralleled with gradual increases in perception of both inclusion within and differentiation from the group. PMID- 15802656 TI - Using images to increase exercise behavior: prototypes versus possible selves. AB - This study investigated the impact of health-related prototypes and possible selves on exercise behavior. In addition, this study examined if these effects might be moderated by individual differences-specifically social comparison (SC) and consideration of future consequences (CFC). One hundred and fifty-two students participated in all three waves of the study-premanipulation, manipulation, and follow-up. As expected, results indicated that individuals who were high on SC and exposed to a prototype manipulation increased their exercise behavior at follow-up. Furthermore, individuals who were future-oriented and exposed to a possible selves manipulation increased their exercise behavior at follow-up. Overall, results revealed that health images have an important impact on health behavior. Implications for behavior change are discussed. PMID- 15802657 TI - The relationship between the need for closure and support for military action against Iraq: moderating effects of national attachment. AB - A variety of studies suggest that a high need for closure--that is, a desire for knowledge that is clear, stable, and unambiguous as opposed to confusing or uncertain--may be associated with greater hostility toward relevant outgroups. Using international attitudes as the context, the authors examine the hypothesis that the relationship between the need for closure and support for military action against Iraq may be moderated by identification with the national ingroup. Specifically, it is expected that this relationship will be moderated by nationalism (i.e., an aggressive form of identification based on a desire for national dominance) but not patriotism (i.e., a more neutral love of one's country). The data provided a clear pattern of support for this hypothesis and additional analyses indicated that a high need for closure reduced variability about the use of force among the highly nationalistic but not the highly patriotic. PMID- 15802658 TI - Group identification and outgroup attitudes in four South African ethnic groups: a multidimensional approach. AB - Although Sumner's ethnocentrism hypothesis, which expects stronger group identification to be associated with more negative outgroup attitudes, has been widely accepted, empirical findings have been inconsistent. This research investigates the relationship of four dimensions of ethnocultural group identification previously proposed by Phinney, that is, salience, evaluation, attachment, and involvement, with attitudes to ethnic outgroups in four South African ethnocultural groups (Africans, Afrikaans Whites, English Whites, Indians). The findings supported the factorial independence of the four identification dimensions and indicated that only one, ethnocultural evaluation (ingroup attitudes), was systematically related to outgroup attitudes, but the association could be positive, negative, or zero. Both functionalist and similarity-dissimilarity approaches to intergroup relations seemed to provide plausible explanations for the pattern of relationships obtained between ingroup and outgroup attitudes. PMID- 15802659 TI - The composition of category conjunctions. AB - In three experiments, the authors investigated the impression formation process resulting from the perception of familiar or unfamiliar social category combinations. In Experiment 1, participants were asked to generate attributes associated with either a familiar or unfamiliar social category conjunction. Compared to familiar combinations, the authors found that when the conjunction was unfamiliar, participants formed their impression less from the individual constituent categories and relatively more from novel emergent attributes. In Experiment 2, the authors replicated this effect using alternative experimental materials. In Experiment 3, the effect generalized to additional (orthogonally combined) gender and occupation categories. The implications of these findings for understanding the processes involved in the conjunction of social categories, and the formation of new stereotypes, are discussed. PMID- 15802660 TI - Self-affirmation and mortality salience: affirming values reduces worldview defense and death-thought accessibility. AB - To the extent that cultural worldviews provide meaning in the face of existential concerns, specifically the inevitability of death, affirming a valued aspect of one's worldview should render reminders of death less threatening. The authors report two studies in support of this view. In Study 1, mortality salience led to derogation of a worldview violator unless participants had first affirmed an important value. In Study 2, self-affirmation before a reminder of death was associated with reduced accessibility of death-related thoughts a short while thereafter. The authors propose that actively affirming one's worldview alters reactions to reminders of mortality by reducing the accessibility of death related thoughts, not by boosting self-esteem. These studies attest to the flexible nature of psychological self-defense and to the central role of cultural worldviews in managing death-related concerns. PMID- 15802661 TI - Myopic biases in strategic social prediction: why deadlines put everyone under more pressure than everyone else. AB - Five experiments document biases in the way people predict the outcomes of interdependent social situations. Participants predicted that situational constraints would restrain their own behavior more than it would the behavior of others, even in situations where everyone faced identical constraints. When anticipating the effects of deadlines on outcomes of negotiations, participants predicted that deadlines would hinder their performance more than it would hinder the performance of others. The results shed light on the psychological processes by which people predict the outcomes of and select strategies in strategic social interaction. They extend prior findings, such as people believing themselves to be below average on difficult tasks, to highly interdependent situations. Furthermore, the article shows both how focusing can account for these effects and also how perspective taking can reduce their biasing influence. PMID- 15802662 TI - Peering into the bias blind spot: people's assessments of bias in themselves and others. AB - People tend to believe that their own judgments are less prone to bias than those of others, in part because they tend to rely on introspection for evidence of bias in themselves but on their lay theories in assessing bias in others. Two empirical consequences of this asymmetry are explored. Studies 1 and 2 document that people are more inclined to think they are guilty of bias in the abstract than in any specific instance. Studies 3 and 4 demonstrate that people tend to believe that their own personal connection to a given issue is a source of accuracy and enlightenment but that such personal connections in the case of others who hold different views are a source of bias. The implications of this asymmetry in assessing objectivity and bias in the self versus others are discussed. PMID- 15802663 TI - Types of high self-esteem and prejudice: how implicit self-esteem relates to ethnic discrimination among high explicit self-esteem individuals. AB - There is increasing recognition that high self-esteem is heterogeneous. Recent research suggests that individuals who report having high self-esteem (i.e., have high explicit self-esteem) behave more defensively to the extent that they have relatively low implicit self-esteem. The current studies test whether individuals with high explicit self-esteem are more likely to discriminate ethnically, as a defensive technique, to the extent that they have relatively low implicit self esteem. The results support this prediction. Among participants with high explicit self-esteem, all of whom were threatened by negative performance feedback, those with relatively low implicit self-esteem recommended a more severe punishment for a Native, but not a White, student who started a fist fight. In Study 2, this pattern was not apparent for participants with relatively low explicit self-esteem. PMID- 15802664 TI - Self-awareness and the emotional consequences of self-discrepancies. AB - Several self theories explore the effects of discrepant self-beliefs on motivation and emotion. This research intersected two self theories: self discrepancy theory and objective self-awareness theory. Self-discrepancy theory predicts that ideal and ought discrepancies cause different negative emotions; objective self-awareness theory predicts that high self-awareness will strengthen the relationship between self-discrepancies and emotions. People (N =112) completed measures of self-discrepancies and emotions (dejection, agitation, positive affect, and negative affect). Self-focused attention was manipulated with a large mirror. When self-awareness was low, self-discrepancies had weak, nonsignificant relations to emotion. When self-awareness was high, however, self discrepancies strongly predicted emotional experience. These effects were general ideal and ought discrepancies affected emotions because of their substantial shared variance, not their unique variance. Implications for theories of self discrepancies and emotions are considered. PMID- 15802665 TI - Evolution of gap junction proteins--the pannexin alternative. AB - Gap junctions provide one of the most common forms of intercellular communication. They are composed of membrane proteins that form a channel that is permeable to ions and small molecules, connecting the cytoplasm of adjacent cells. Gap junctions serve similar functions in all multicellular animals (Metazoa). Two unrelated protein families are involved in this function; connexins, which are found only in chordates, and pannexins, which are ubiquitous and present in both chordate and invertebrate genomes. The involvement of mammalian pannexins to gap junction formation was recently confirmed. Now it is necessary to consider the role of pannexins as an alternative to connexins in vertebrate intercellular communication. PMID- 15802666 TI - Prey-capture in Pomacanthus semicirculatus (Teleostei, Pomacanthidae): functional implications of intramandibular joints in marine angelfishes. AB - We examined prey-capture morphology and kinematics in the angelfish, Pomacanthus semicirculatus (Cuvier 1931), to evaluate the magnitude and role of functional specialisation. The feeding apparatus of P. semicirculatus possess three biomechanical mechanisms of particular interest: (1) a novel intramandibular joint, permitting dentary rotation and protruded jaw closure; (2) an opercular linkage facilitating mandible depression; and (3) a suspensorial linkage with two novel points of flexion, permitting anterior rotation of the suspensorium and augmenting mandible protrusion. Prey-capture kinematics were quantified using motion analysis of high-speed video, yielding performance profiles illustrating timing of onset, duration and magnitude of movement in these three biomechanical systems, and other variables traditionally quantified in studies of teleostean ram-suction feeding activity. Mandible depression and suspensorial rotation both augmented mandible protrusion, and coincided during jaw protrusion, typically increasing head length by 30%. Jaw closure appeared to result from contraction of the adductor mandibulae segment A2, which rotated the dentary by approximately 30 degrees relative to the articular. This resulted in jaw closure with the mandible fully depressed and the jaws at peak-protrusion. Feeding events were concluded by a high-velocity jaw retraction (20-50 ms), and completed in 450-750 ms. Feeding kinematics and morphology of Pomacanthus differed from other biting teleosts, and more closely resemble some long-jawed ram-suction feeders. The structural and functional modifications in the Pomacanthus feeding apparatus are matched to an unusual diet of structurally resilient and firmly attached benthic prey. PMID- 15802667 TI - Propulsive force calculations in swimming frogs. I. A momentum-impulse approach. AB - Frogs are animals that are capable of locomotion in two physically different media, aquatic and terrestrial. A comparison of the kinematics of swimming frogs in a previous study revealed a difference in propulsive impulse between jumping and swimming. To explore this difference further, we determined the instantaneous forces during propulsion in swimming using an impulse-momentum approach based on DPIV flow data. The force profile obtained was compared with force profiles obtained from drag-thrust equilibrium of the centre of mass and with the force profiles generated during jumping. The new approach to quantifying the instantaneous forces during swimming was tested and proved to be a valid method for determining the external forces on the feet of swimming frogs. On the kinematic profiles of swimming, leg extension precedes propulsion. This means that it is not only the acceleration of water backwards that provides thrust, but also that the deceleration of water flowing towards the frog as a result of recovery accelerates the centre of mass prior to leg extension. The force profile obtained from the impulse-momentum approach exposed an overestimation of drag by 30% in the drag-thrust calculations. This means that the difference in impulse between jumping and swimming in frogs is even larger than previously stated. The difference between the force profiles, apart from a slightly higher peak force during jumping, lies mainly in a difference in shape. During swimming, maximal force is reached early in the extension phase, 20% into it, while during jumping, peak force is attained at 80% of the extension phase. This difference is caused by a difference in inter-limb coordination. PMID- 15802668 TI - Propulsive force calculations in swimming frogs. II. Application of a vortex ring model to DPIV data. AB - Frogs propel themselves by kicking water backwards using a synchronised extension of their hind limbs and webbed feet. To understand this propulsion process, we quantified the water movements and displacements resulting from swimming in the green frog Rana esculenta, applying digital particle image velocimetry (DPIV) to the frog's wake. The wake showed two vortex rings left behind by the two feet. The rings appeared to be elliptic in planform, urging for correction of the observed ring radii. The rings' long and short axes (average ratio 1.75:1) were about the same size as the length and width of the propelling frog foot and the ellipsoid mass of water accelerated with it. Average thrust forces were derived from the vortex rings, assuming all propulsive energy to be compiled in the rings. The calculated average forces (F(av)=0.10+/-0.04 N) were in close agreement with our parallel study applying a momentum-impulse approach to water displacements during the leg extension phase. We did not find any support for previously assumed propulsion enhancement mechanisms. The feet do not clap together at the end of the power stroke and no "wedge-action" jetting is observed. Each foot accelerates its own water mantle, ending up in a separate vortex ring without interference by the other leg. PMID- 15802669 TI - The role of ultraviolet wavelengths in the mate-choice decisions of female three spined sticklebacks. AB - Female three-spined sticklebacks have been found to use visual cues when responding sexually towards courting males, often preferring more intensely red coloured males, and males with blue rather than silver irises. However, traditionally the literature has failed to test preference across the full spectral range to which females might be sensitive, limiting analysis to the human-visible wavelengths of the electromagnetic spectrum. We studied the effects that the addition of ultraviolet wavelengths has on the mate-choice preferences of female sticklebacks using a two-choice paradigm. We found that females preferred males that were viewed across the full spectrum to males whose display lacked an ultraviolet component. Using suitable controls we were able to establish that female preference was sexually motivated and was not caused by a general preference for the manipulated light conditions. Our results indicate that female preference may be due to an enhancement in visual contrast when males are viewed in full spectrum conditions. PMID- 15802670 TI - Juvenile Lymnaea ventilate, learn and remember differently than do adult Lymnaea. AB - Adult snails are capable of learning associatively not to perform aerial respiration and then to consolidate the acquired behaviour into long-term memory (LTM). Juvenile Lymnaea, however, perform aerial respiration significantly less often and the three-neuron circuit that drives this behaviour operates significantly differently than in it does in adults. We asked whether these ontogenic behavioural and neurophysiological differences are manifested as an altered ability of juveniles to learn and/or form LTM. We found that juvenile snails learn significantly less well than adults and are, as a group, incapable of forming LTM. To control for the possibility that the poor learning and inability to form memory were the result of juvenile's receiving on average fewer reinforcing stimuli because they perform aerial respiration less often than adults we subjected juveniles to an enforced period of hypoxia to "motivate" juveniles. Motivated juveniles perform aerial respiration as often as adults; yet these "motivated" juveniles continue to be poor learners and still cannot form LTM. Additionally, a small percentage of juveniles perform aerial respiration as often as adults (i.e. high responders). When these "high-responders" were trained they still exhibited poorer learning ability compared with adults and could not form LTM. We conclude that juvenile snails have a more difficult time learning and remembering to suppress aerial respiratory activity than do adults. PMID- 15802671 TI - The ontogenetic changes in the thermal properties of blubber from Atlantic bottlenose dolphin Tursiops truncatus. AB - In Atlantic bottlenose dolphins Tursiops truncatus, both the thickness and lipid content of blubber vary across ontogeny and across individuals of differing reproductive and nutritional status. This study investigates how these changes in blubber morphology and composition influence its thermal properties. Thermal conductivity (W m(-1) deg.(-1), where deg. is degrees C) and thermal insulation (m(2) deg. W(-1)) of dolphin blubber were measured in individuals across an ontogenetic series (fetus through adult, N=36), pregnant females (N=4) and emaciated animals (N=5). These thermal properties were determined by the simultaneous use of two common experimental approaches, the heat flux disc method and the standard material method. Thickness, lipid and water content were measured for each blubber sample. Thermal conductivity and insulation varied significantly across ontogeny. Blubber from fetuses through sub-adults was less conductive (range=0.11-0.13+/-0.02 W m(-1) deg.(-1)) than that of adults (mean=0.18 W m(-1) deg.(-1)). The conductivity of blubber from pregnant females was similar to non-adult categories, while that of emaciated animals was significantly higher (0.24 +/- 0.04 W m deg.(-1)) than all other categories. Blubber from sub-adults and pregnant females had the highest insulation values while fetuses and emaciated animals had the lowest. In nutritionally dependent life history categories, changes in blubber's thermal insulation were characterized by stable blubber quality (i.e. conductivity) and increased blubber quantity (i.e. thickness). In nutritionally independent animals, blubber quantity remained stable while blubber quality varied. A final, unexpected observation was that heat flux measurements at the deep blubber surface were significantly higher than that at the superficial surface, a pattern not observed in control materials. This apparent ability to absorb heat, coupled with blubber's fatty acid composition, suggest that dolphin integument may function as a phase change material. PMID- 15802672 TI - Tuning of host plants with vibratory songs of Nezara viridula L (Heteroptera: Pentatomidae). AB - Songs of the southern green stink bug Nezara viridula L, recorded on a non resonant loudspeaker membrane, reflect frequency characteristics of body vibrations. The song dominant frequency directly depends on the repetition rate of potentials recorded from synchronously contracting muscles that vibrate the abdomen during singing. Spectra of naturally emitted signals recorded on the pronotum of a singing bug or on a plant contain peaks characteristic of plant resonant spectra. The dominant resonant frequency of sound-induced vibrations in bean and other stink bug host plants ranges between 160 and 215 Hz and subdominant peaks do not exceed 600 Hz. There is no correlation between spectral peak positions and recording points on a plant. The dominant resonance peak corresponds to the best frequency sensitivity of N. viridula middle frequency subgenual receptor cell. Subdominant peaks around and below 100 Hz lie close to the dominant frequency of body vibrations during singing and to the range of best frequency sensitivity of low frequency receptor cells. Tuning of plant resonant frequencies with spectral properties of songs and frequency sensitivity of sensory organs is discussed in the context of stink bug substrate-borne communication. PMID- 15802673 TI - Giant Galapagos tortoises walk without inverted pendulum mechanical-energy exchange. AB - Animals must perform mechanical work during walking, but most conserve substantial mechanical energy via an inverted-pendulum-like mechanism of energy recovery in which fluctuations of kinetic energy (KE) and gravitational potential energy (GPE) are of similar magnitude and 180 degrees out of phase. The greatest energy recovery typically occurs at intermediate speeds. Tortoises are known for their slow speeds, which we anticipated would lead to small fluctuations in KE. To have an effective exchange of mechanical energy using the inverted-pendulum mechanism, tortoises would need to walk with only small changes in GPE corresponding to vertical center-of-mass (COM) fluctuations of < 0.5 mm. Thus, we hypothesized that giant Galapagos tortoises would not conserve substantial mechanical energy using the inverted-pendulum mechanism. We studied five adult giant Galapagos tortoises Geochelone elephantopus (mean mass=142 kg; range= 103 196 kg). Walking speed was extremely slow (0.16+/-0.052 m s(-1); mean +/- 1 s.d.). The fluctuations in kinetic energy (8.1+/-3.98 J stride(-1)) were only one third as large as the fluctuations in gravitational potential energy (22.7+/-8.04 J stride(-1)). In addition, these energies fluctuated nearly randomly and were only sporadically out of phase. Because of the dissimilar amplitudes and inconsistent phase relationships of these energies, tortoises conserved little mechanical energy during steady walking, recovering only 29.8+/-3.77% of the mechanical energy (range=13-52%). Thus, giant Galapagos tortoises do not utilize effectively an inverted-pendulum mechanism of energy conservation. Nonetheless, the mass-specific external mechanical work required per distance (0.41+/-0.092 J kg(-1) m(-1)) was not different from most other legged animals. Other turtle species use less than half as much metabolic energy to walk as other terrestrial animals of similar mass. It is not yet known if Galapagos tortoises are economical walkers. Nevertheless, contrary to biomechanical convention, poor inverted-pendulum mechanics during walking do not necessarily correspond to high mechanical work and may not result in a high metabolic cost. PMID- 15802674 TI - Simultaneous measurement of metabolic and acoustic power and the efficiency of sound production in two mole cricket species (Orthoptera: Gryllotalpidae). AB - We here report the first simultaneous measurement of metabolic cost of calling, acoustic power and efficiency of sound production in animals--the mole crickets Scapteriscus borellii and S. vicinus (Gryllotalpidae). We measured O(2) consumption, CO(2) production and acoustic power as the crickets called from their burrows in an open room. We utilized their calling burrow as the functional equivalent of a mask. Both species had a respiratory quotient near 0.85, indicative of metabolism based on a mix of carbohydrates and fats. The metabolic rate was significantly higher in S. borellii (11.6 mW g(-1)) than in S. vicinus (9.0 mW g(-1)) and averaged about eight- to fivefold greater, respectively, than resting metabolism. In some individuals, metabolic rate decreased by 20% during calling bouts. Costs of refurbishing calling burrows in S. borellii were less than calling costs, due to the behavior's short duration (ca. 15 min) and its relatively low average metabolic rate (4 mW). Acoustic power was on average sevenfold greater in S. borellii (21.2 vs 2.9 microW) and was more variable within individuals and across species than the metabolic rate. The efficiency of sound production was significantly higher in S. borellii (0.23 vs 0.03%). These values are below published estimates for other insects even though these mole crickets construct acoustic burrows that have the potential to increase efficiency. The cricket/burrow system in both species have an apparent Q(ln decrement) of about 6, indicative of significant internal damping caused by the airspaces in the sand that forms the burrow's walls. Damping is therefore an important cause of the low sound production efficiency. In field conditions where burrow walls are saturated with water and there is less internal damping, calls are louder and sound production efficiency is likely higher. File tooth depths and file tooth-to-tooth distances correlated with interspecific differences in metabolism and acoustic power much better than with wing stroke rates and plectrum-to-file tooth strike rates. To further investigate these correlations, we constructed two models of energy input to the tegminal oscillator. A model based on transfer of kinetic energy based on differences in tegminal velocity and file tooth spacing showed the most promise. Related calculations suggest that if there are no elastic savings, the power costs to accelerate and decelerate the tegmina are greater than the predicted power input to the tegminal oscillator, and that they are similar in the two species even though S. vicinus has a nearly threefold higher wing stroke rate. PMID- 15802675 TI - A new model predicting locomotor cost from limb length via force production. AB - Notably absent from the existing literature is an explicit biomechanical model linking limb design to the energy cost of locomotion, COL. Here, I present a simple model that predicts the rate of force production necessary to support the body and swing the limb during walking and running as a function of speed, limb length, limb proportion, excursion angle and stride frequency. The estimated rate of force production is then used to predict COL via this model following previous studies that have linked COL to force production. To test this model, oxygen consumption and kinematics were measured in nine human subjects while walking and running on a treadmill at range of speeds. Following the model, limb length, speed, excursion angle and stride frequency were used to predict the rate of force production both to support the body's center of mass and to swing the limb. Model-predicted COL was significantly correlated with observed COL, performing as well or better than contact time and Froude number as a predictor of COL for running and walking, respectively. Furthermore, the model presented here predicts relationships between COL, kinematic variables and body size that are supported by published reduced-gravity experiments and scaling studies. Results suggest the model is useful for predicting COL from anatomical and kinematic variables, and may be useful in intra- and inter-specific studies of locomotor anatomy and performance. PMID- 15802676 TI - Boosting intermediate-term into long-term memory. AB - Aerial respiration in the pond snail Lymnaea stagnalis can be operantly conditioned. Depending on the specific training procedure used (i.e. a 0.5 h vs a 1.0 h interval between training sessions) either intermediate (ITM) or long-term memory (LTM) is formed. ITM, which persists for 2-3 h, is dependent only on de novo protein synthesis, whilst LTM persists for up to 4 weeks and is dependent on both transcription and de novo protein synthesis. We found that although the behavioural phenotype of ITM was not apparent 24 h after the last training session, a residual memory trace was present that serves as a foundation upon which a subsequent ITM-training-procedure builds on to form LTM (i.e. a "changed memory"). This residual memory trace could be perturbed by cooling, the behavioural process of context-specific extinction and by increasing the interval between the training procedures. Furthermore in preparations where the somata of RPeD1 (one of three interneurons in the central pattern generator required for aerial respiratory behavior) had been ablated before training, LTM could not be observed following a second bout of ITM-training. These data support the concept that a molecular memory trace is established as a consequence of ITM-training, which serves as a "permissive substrate", when the ITM memory is made active, sufficient to permit the necessary transcription and translation processes that are causal for LTM formation. PMID- 15802677 TI - Development of oxygen sensing in the gills of zebrafish. AB - Previous studies have described the morphology, innervation and O(2) chemoreceptive properties of neuroepithelial cells (NECs) of the zebrafish gill filaments. The present work describes the ontogenesis of these cells, and the formation of functional O(2)-sensing pathways in developing zebrafish. Confocal immunofluorescence was performed on whole-mount gill preparations using antibodies against serotonin (5-HT) and a zebrafish-derived neuronal marker (zn 12) to identify the appearance and innervation of gill NECs during larval stages. NECs were first expressed in gill filament primordia of larvae at 5 days postfertilization (d.p.f.) and were fully innervated by 7 d.p.f. In vivo ventilation frequency analysis revealed that a behavioural response to hypoxia (11.2+/-2.8 min(-1)) developed in embryos as early as 2 d.p.f., and a significant increase (P<0.05) in the ventilatory response to hypoxia (200.8+/-23.0 min(-1)) coincided with innervation of NECs of the filaments. In addition, exogenous application of quinidine, a blocker of O(2)-sensitive background K(+) channels in NECs, induced hyperventilation in adults in a dose-dependent manner and revealed the development of a quinidine-sensitive ventilatory response in 7 d.p.f. larvae. This study shows that NEC innervation in the gill filaments may account for the development of a functional O(2)-sensing pathway and the hyperventilatory response to hypoxia in zebrafish larvae. At earlier stages, however, O(2)-sensing must occur through another pathway. The possibility that a new type of 5-HT positive NEC of the gill arches may account for this earlier hypoxic response is discussed. PMID- 15802678 TI - Influence of environmental conditions on early development of the hydrothermal vent polychaete Alvinella pompejana. AB - Dispersal and colonisation processes at deep-sea vents are still not fully understood, essentially because early life stages of vent species remain unknown. The polychaete worm Alvinella pompejana forms colonies on chimney walls at East Pacific Rise vent sites where the temperature can frequently exceed 20 degrees C. In vitro studies in pressure vessels showed that the early embryos tolerate temperatures in a lower range (10-14 degrees C), suggesting that they would have to escape the colony to develop. Pressure vessels offer the advantage that each parameter can be independently controlled, but they do not simulate the more complex and dynamic conditions naturally encountered at vent sites. Accordingly, in addition to incubations in pressure vessels, we incubated embryos directly at a vent site, in different habitats along a gradient of hydrothermal influence. Embryos incubated on an adult A. pompejana colony where temperature and H(2)S concentrations were relatively high showed a very low survival rate and did not develop, whereas embryos incubated in a Riftia pachyptila clump environment with a lower hydrothermal signature, or at the base of the chimney where the influence of the hydrothermal activity was very weak, survived well and developed. Although the average temperature recorded in the A. pompejana colony was within the range tolerated by embryos (13 degrees C), frequent peaks above 20 degrees C were recorded. Estimated sulphide concentration at this site reached 200 mumol l(-1). Punctuated exposure to both high temperature and elevated sulphide levels probably explain the low survival of embryos within the A. pompejana colony. The in situ experiments further support the idea that embryos require conditions with moderate hydrothermal influence not generally found within an adult colony. However, as much more benign physicochemical conditions can be found within a few tens of cm of adult colonies, embryos do not necessarily have to leave their vent of origin to develop. Further analyses are needed to pinpoint the specific factors that affect the survival and development of embryos at vents. PMID- 15802679 TI - A single control system for smooth and saccade-like pursuit in blowflies. AB - During courtship, male blowflies perform aerobatic pursuits that rank among the fastest visual behaviours that can be observed in nature. The viewing strategies during pursuit behaviour of blowflies appear to be very similar to eye movements during pursuit in primates: a combination of smooth pursuit and catch-up saccades. Whereas in primates these two components of pursuit eye movements are thought to be controlled by distinct oculomotor subsystems, we present evidence that in blowflies both types of pursuit responses can be produced by a single control system. In numerical simulations of chasing behaviour the proposed control system generates qualitatively the same behaviour as with real blowflies. As a consequence of time constants in the control system, mimicking neuronal processing times, muscular dynamics and inertia, saccade-like changes in gaze direction are generated if the target is displaced rapidly on the pursuing fly's retina. In the behavioural context of visual pursuit, saccade-like changes of the fly's gaze direction can thus be parsimoniously explained as an emergent property of a smooth pursuit system without assuming a priori different mechanisms underlying smooth and saccadic tracking behaviour. PMID- 15802680 TI - The neglect of volition. PMID- 15802681 TI - Essential fatty acids and mental health. PMID- 15802682 TI - Dialectical behaviour therapy in the treatment of borderline personality disorder. PMID- 15802683 TI - Pathways to care and ethnicity. 1: Sample characteristics and compulsory admission. Report from the AESOP study. AB - BACKGROUND: Many studies have found high levels of compulsory admission to psychiatric hospital in the UK among African-Caribbean and Black African patients with a psychotic illness. AIMS: To establish whether African-Caribbean and Black African ethnicity is associated with compulsory admission in an epidemiological sample of patients with a first episode of psychosis drawn from two UK centres. METHOD: All patients with a first episode of psychosis who made contact with psychiatric services over a 2-year period and were living in defined areas were included in the (AESOP) study. For this analysis we included all White British, other White, African-Caribbean and Black African patients from the AESOP sampling frame. Clinical, socio-demographic and pathways to care data were collected from patients, relatives and case notes. RESULTS: African-Caribbean patients were significantly more likely to be compulsorily admitted than White British patients, as were Black African patients. African-Caribbean men were the most likely to be compulsorily admitted. CONCLUSIONS: These findings suggest that factors are operating at or prior to first presentation to increase the risk of compulsory admission among African-Caribbean and Black African patients. PMID- 15802684 TI - Pathways to care and ethnicity. 2: Source of referral and help-seeking. Report from the AESOP study. AB - BACKGROUND: Previous research has found that African-Caribbean and Black African patients are likely to come into contact with mental health services via more negative routes, when compared with White patients. We sought to investigate pathways to mental health care and ethnicity in a sample of patients with a first episode of psychosis drawn from two UK centres. METHOD: We included all White British, other White, African-Caribbean and Black African patients with a first episode of psychosis who made contact with psychiatric services over a 2-year period and were living in defined areas. Clinical, socio-demographic and pathways to care data were collected from patients, relatives and case notes. RESULTS: Compared with White British patients, general practitioner referral was less frequent for both African-Caribbean and Black African patients and referral by a criminal justice agency was more common. With the exception of criminal justice referrals for Black African patients, these findings remained significant after adjusting for potential confounders. CONCLUSIONS: These findings suggest that factors are operating during a first episode of psychosis to increase the risk that the pathway to care for Black patients will involve non-health professionals. PMID- 15802685 TI - Help-seeking behaviour in men and women with common mental health problems: cross sectional study. AB - BACKGROUND: Many people with mental health problems do not seek professional help but their use of other sources of help is unclear. AIMS: To investigate patterns of lay and professional help-seeking in men and women aged 16-64 years in relation to severity of symptoms and socio-demographic variables. METHOD: Postal questionnaire survey, including the 12-item General Health Questionnaire (GHQ 12), sent to a stratified random sample (n=15,222) of the population of Somerset. RESULTS: The response rate was 76%. Only 28% of people with extremely high GHQ-12 scores (> or = 8) had sought help from their general practitioner but most (78%) had sought some form of help. Males, young people and people living in affluent areas were the least likely to seek help. CONCLUSIONS: Health promotion interventions to encourage appropriate help-seeking behaviour in young people, particularly in men, may lead to improvements in the mental health of this group of the population. PMID- 15802686 TI - Negative self-schemas and the onset of depression in women: longitudinal study. AB - BACKGROUND: Beck's cognitive theory of depression has received little empirical support. AIMS: To test whether those with negative self-schemas were at risk of onset of depression. METHOD: Data were collected by postal questionnaire from 12,003 women recruited during early pregnancy; questionnaires included measures of depressive symptoms and negative self-schemas. Regular questionnaires were sent during pregnancy and following childbirth. RESULTS: Of 8540 women not depressed when recruited, 8.6% (95% CI 8.0-9.2) became depressed 14 weeks later. Those in the highest tertile for negative self-schema score were more likely to become depressed than those in the lowest tertile (odds ratio 3.04, 95% CI 2.48 3.73). The association remained after adjustment for baseline depressive symptoms and previous depression (OR 1.6, 95% CI 1.27-2.02) and was of similar magnitude for onset 3 years later. CONCLUSIONS: Holding a negative self-schema is an independent risk factor for the onset of depression in women. This finding supports a key element of Beck's cognitive theory. Understanding more about how negative self-schemas arise should help inform preventive policies. PMID- 15802687 TI - Neurological findings in late-onset depressive disorder: comparison of individuals with and without depression. AB - BACKGROUND: Organic factors are thought to be important in late-life depressive disorder but there have been few studies specifically of neurological signs. AIMS: To compare neurological signs in a group of patients with late-onset depression and in healthy controls. METHOD: A case-control study comparing 50 patients with depression and 35 controls on three measures of central nervous system (CNS) signs: a structured CNS examination, the Neurological Evaluation Scale (NES) and the Webster rating scale for parkinsonism. RESULTS: After adjusting for major depression at the time of evaluation and prescription of tranquillisers, ratings on two of the NES sub-scales (complex motor sequencing and 'other' signs) and on the Webster scale were significantly higher (more impaired) in patients compared with controls (P<0.05). With logistic regression, the NES was the main measure predictive of group outcome. There were no differences in scores of vascular risk or white matter but patients had patients had more atrophy. CONCLUSIONS: The findings add to the evidence that late-life depression is associated with organic brain dysfunction, perhaps mediated by neurodegeneration or subtle vascular impairment. The use of the NES in subjects with depression should be replicated. PMID- 15802688 TI - Prospective study of risk factors for attempted suicide among patients with DSM IV major depressive disorder. AB - BACKGROUND: There are few prospective studies on risk factors for attempted suicide among psychiatric out- and in-patients with major depressive disorder. AIMS: To investigate risk factors for attempted suicide among psychiatric out- and in-patients with major depressive disorder in the city of Vantaa, Finland. METHOD: The Vantaa Depression Study included 269 patients with DSM-IV major depressive disorder diagnosed using semistructured interviews and followed up at 6- and 18-month interviews with a life chart. RESULTS: During the 18-month followup, 8% of the patients attempted suicide. The relative risk of an attempt was 2.50 during partial remission and 7.54 during a major depressive episode, compared with full remission (P 0.001). Numerous factors were associated with this risk, but lacking a partner, previous suicide attempts and total time spent in major depressive episodes were the most robust predictors. CONCLUSIONS: Suicide attempts among patients with major depressive disorder are strongly associated with the presence and severity of depressive symptoms and predicted by lack of partner, previous suicide attempts and time spent in depression. Reducing the time spent depressed is a credible preventive measure. PMID- 15802689 TI - The PAS-ADD Checklist: independent replication of its psychometric properties in a community sample. AB - BACKGROUND: The development of reliable, valid measures of psychopathology in people with intellectual disabilities is an important task. However, independent replication studies are rarely reported. AIMS: To report data on the psychometric properties of the Psychiatric Assessment Schedule for Adults with Developmental Disabilities (PAS-ADD) Checklist. METHOD: The PAS-ADD Checklist was completed for 226 adults as part of the assessment process for a specialist mental health service for people with intellectual disabilities. RESULTS: Internal consistency was acceptable. Factor analysis revealed one main factor that was characterised by items related to mood. The Checklist was sensitive to differences between diagnostic groups and had an overall sensitivity of 66%; its specificity was 70%. CONCLUSIONS: The PAS-ADD Checklist is a quick and easy to use screening tool. Although at present it is the best measure available, it should not be the only method used to identify psychiatric disorders in people with intellectual disabilities. PMID- 15802690 TI - Cognitive-behavioural therapy for refractory psychotic symptoms of schizophrenia resistant to atypical antipsychotic medication. Randomised controlled trial. AB - BACKGROUND: There is increasing evidence that cognitive-behavioural therapy can be an effective intervention for patients experiencing drug-refractory positive symptoms of schizophrenia. AIMS: To investigate the effects of cognitive behavioural therapy on in-patients with treatment-refractory psychotic symptoms. METHOD: Manualised therapy was compared with supportive counselling in a randomised controlled study. Both interventions were delivered by experienced psychologists over 16 sessions of treatment. Therapy fidelity was assessed by two independent raters. Participants underwent masked assessment at baseline, after treatment and at 6 months' follow-up. Main outcome measures were the Positive and Negative Syndrome Scale and the Psychotic Symptoms Rating Scale. The analysis was by intention to treat. RESULTS: Participants receiving cognitive cognitive behavioural therapy had improved with regard to auditory hallucinations and illness insight at the post-treatment assessment, but these findings were not maintained at follow-up. CONCLUSIONS: Cognitive-behavioural therapy showed modest short-term benefits over supportive counselling for treatment-refractory positive symptoms of schizophrenia. PMID- 15802691 TI - Causal beliefs and attitudes to people with schizophrenia. Trend analysis based on data from two population surveys in Germany. AB - BACKGROUND: It is a widely shared belief that an increase in mental health literacy will result in an improvement of attitudes towards people with mental illness. AIMS: To examine how the German public's causal attributions of schizophrenia and their desire for social distance from people with schizophrenia developed over the 1990s. METHOD: A trend analysis was carried out using data from two representative population surveys conducted in the Lander constituting the former Federal Republic of Germany in 1990 and 2001. RESULTS: Parallel to an increase in the public's tendency to endorse biological causes, an increase in the desire for social distance from people with schizophrenia was found. CONCLUSIONS: The assumption underlying current anti-stigma programmes that there is a positive relationship between endorsing biological causes and the acceptance of people with mental illness appears to be problematic. PMID- 15802692 TI - Assessing psychopathy in the UK: concerns about cross-cultural generalisability. AB - BACKGROUND: The diagnosis of psychopathy is important for violence risk assessment. AIMS: To investigate whether the syndromal structure of psychopathy, as measured by the Psychopathy Checklist-Revised (PCL-R), is the same in the UK and North America, and whether this measure yields scores that are equivalent in these two regions. METHOD: Confirmatory factor analytic and item response theory methods were applied to large samples of PCL-R ratings. RESULTS: The syndromal structure of psychopathy was invariant across cultures, three distinct factors underpinning the superordinate syndrome of psychopathy. However, PCL-R scores were not equivalent across cultures: the same level of psychopathy was associated with lower PCL-R scores in the UK. Items that reflected affective symptoms had the highest cross-cultural stability. CONCLUSIONS: Scores on the PCL-R obtained in the UK are not directly comparable with those obtained in North America. Care must be exercised when the PCL-R is used to make important clinical decisions in the UK. PMID- 15802693 TI - Transference patterns in the psychotherapy of personality disorders: empirical investigation. AB - BACKGROUND: The concept of transference has broadened to a recognition that patients often express enduring relational patterns in the therapeutic relationship. AIMS: To examine the structure of patient relational patterns in psychotherapy and their relation with DSM-IV personality disorder symptoms. METHOD: A random sample of psychologists and psychiatrists (n=181) completed a battery of instruments on a randomly selected patient in their care. RESULTS: Exploratory factor analysis identified five transference dimensions: angry/entitled, anxious/preoccupied, avoidant/counterdependent, secure/engaged and sexualised. These were associated in predictable ways with Axis II pathology; four mapped on to adult attachment styles. An aggregated portrait of transference patterns in narcissistic patients provided a clinically rich, empirically based description of transference processes that strongly resembled clinical theories. CONCLUSIONS: The ways patients interact with their therapists can provide important data about their personality, attachment patterns and interpersonal functioning. These processes can be measured in clinically sophisticated and psychometrically sound ways. Such processes are relatively independent of clinicians' theoretical orientation. PMID- 15802694 TI - Predictors of outcome following treatment for chronic fatigue. AB - We explored the role of baseline characteristics of 105 patients who presented with fatigue in primary care in determining outcome following either graded exercise or cognitive-behavioural therapy. Meeting the criteria for chronic fatigue syndrome was the most powerful predictor of poor outcome and this negative effect was enhanced by greater functional impairment or greater perceived negative consequences, but was not further enhanced by both. PMID- 15802695 TI - Occult suicidality in an emergency department population. AB - The prevalence and severity of suicidal ideation was established in a patient sample seeking emergency treatment for non-psychiatric reasons. Using a computerised mental health screening panel, data were collected from waiting-room patients during randomised shifts over a 45-day period. Of 1590 screened patients, 185 (11.6%) acknowledged suicidal ideation and 31 (2%) reported planning to kill themselves. Almost all of those with suicidal ideation (97%) acknowledged symptoms consistent with mood, anxiety and/or substance-related disorders. Structured medical record review revealed that 25 of the 31 patients planning suicide were undetected during their index visit, and that 4 attempted suicide within 45 days of the visit. All survived. PMID- 15802696 TI - Suicide in prisons. PMID- 15802698 TI - Attention deficit disorder in adults. PMID- 15802697 TI - Partial agonists in schizophrenia. PMID- 15802699 TI - Diagnosis of coeliac disease. PMID- 15802700 TI - What makes a good clinical decision support system. PMID- 15802701 TI - Meeting the ethical needs of doctors. PMID- 15802702 TI - Children with psychiatric disorders and learning disabilities. PMID- 15802703 TI - A wolf in sheep's clothing. PMID- 15802704 TI - Walk-in centres fail to take pressure off emergency services in England. PMID- 15802705 TI - Parents should have right to choose sex of child, say MPs. PMID- 15802706 TI - Europe is a "second class continent" for cancer research. PMID- 15802707 TI - US Supreme Court refuses to intervene in "right to die" case. PMID- 15802708 TI - Draft mental health bill needs major overhaul, says committee. PMID- 15802709 TI - Dutch Supreme Court backs damages for child for having been born. PMID- 15802712 TI - Indian proposals to revalidate doctors get mixed reception. PMID- 15802713 TI - Consumers' organisation raises questions over choice in the NHS. PMID- 15802714 TI - Treatment of prostate cancer lags behind other cancers. PMID- 15802715 TI - Regular inspections of NHS trusts to be dropped, in favour of self reporting. PMID- 15802716 TI - FDA seeks to ease burden on trial review boards. PMID- 15802717 TI - Preventable infections are out of control in Canadian hospitals. PMID- 15802718 TI - Open access publishing: too much oxygen? PMID- 15802719 TI - Recent developments in bisphosphonates for patients with metastatic breast cancer. PMID- 15802720 TI - Atypical presentation of coeliac disease. PMID- 15802722 TI - Substance misuse: alcohol, tobacco, inhalants, and other drugs. PMID- 15802721 TI - Antibody negative coeliac disease presenting in elderly people--an easily missed diagnosis. PMID- 15802723 TI - Five pitfalls in decisions about diagnosis and prescribing. PMID- 15802724 TI - Can we avoid bias? PMID- 15802725 TI - Surgery is the best intervention for severe coronary artery disease. PMID- 15802726 TI - Moving towards true integration. PMID- 15802727 TI - Cognitive behaviour therapy for adolescents with chronic fatigue syndrome: data are insufficient and conclusion inappropriate. PMID- 15802728 TI - Adolescent development: advice in ABC of adolescence is potentially misleading. PMID- 15802729 TI - Clomipramine and neuroleptic malignant syndrome: literature on adverse reactions to psychotropic drugs continues to confuse. PMID- 15802731 TI - How to prevent caesarean deliveries deserves more study. PMID- 15802732 TI - Need for expertise based randomised controlled trials: expertise based design has shortfalls. PMID- 15802733 TI - Risks of gene therapy should be weighed against lack of alternatives for many diseases. PMID- 15802734 TI - Old docs and new tricks: Efficiency is important. PMID- 15802735 TI - Old docs and new tricks: seasoned doctors may be better than young doctors at some things. PMID- 15802736 TI - Target SHOs and registrars for communication skills training. PMID- 15802737 TI - Need for expertise based randomised controlled trials: surgical research shares many similarities with psychotherapy research. PMID- 15802739 TI - The concept of intentionality in human science nursing theories. AB - Nursing as a human science focuses on life and health as humanly experienced. Intentionality is a philosophical idea of particular significance to nursing as a human science, particularly within Parse's theory of human becoming and Watson's theory of human caring. These two theorists' interpretations of intentionality are explored in relation to their respective theories and the implications for research and practice. PMID- 15802740 TI - Feeling respected-not respected: the embedded artist in Parse method research. AB - The purpose of this column is to present the plan of a research project involving researchers, artists, and research participants. The planned research project will explore with the Parse method eight universal lived experiences important to persons in community. It is anticipated that art works will complement story and text in order to enhance understanding. The authors here present the first attempt of including an embedded artist with Parse method involving one participant who spoke about her experience of feeling respected-not respected. Researcher text and artist works are shown to enhance meaning and understanding. PMID- 15802742 TI - Scholarship in nursing: ethics of a practice doctorate. AB - The American Association of Colleges of Nursing recently adopted a position statement which gave approval for the development of a doctorate of nursing practice. What are possible ethics related to meaning and scholarship in providing this form of terminal degree in nursing? What are some questions that members of the discipline ought to be thinking about when pondering beginning such programs? When viewed from a nursing theoretical perspective, what are potential implications for initiating this degree? This column shall begin the discussion regarding the ethics and possible meanings for doing what is best for the discipline of nursing. PMID- 15802743 TI - A community of scholars: what is it? AB - This column addresses the question, What is a community of scholars? This question is explored through the writings of authors about genius--looking at such geniuses as Plato, Mahatma Ghandi, Queen Elizabeth I, and Christopher Columbus. The question is also explored through the writings of present day nurse scholars and their meanings of community of scholars. PMID- 15802744 TI - The language of loss when a baby dies prior to birth: cocreating human experience. AB - When a baby dies prior to birth, a woman and her family begin a life long journey of living and transforming with loss. The language used with families during times of devastating loss is important to reflect upon since, language has the potential to either intensify suffering or enhance the family's experience of grieving. Words that affirm the meaning the baby holds for the family can provide comfort as families engage with the reality of lost hopes and dreams. Recognizing that healthcare professionals do not intend to harm families whose babies die prior to birth, the current authors offer this column as a vehicle for reflecting on the meanings of words used during this particular time of loss and grief. The authors further explore the role that the arts have to play in expressing loss and in helping others to understand. PMID- 15802745 TI - Criteria for evaluation of theory. AB - This column presents criteria for evaluation of nursing theories specified by Jacqueline Fawcett and Rosemarie Rizzo Parse. Fawcett's criteria are significance, internal consistency, parsimony, testability, empirical adequacy, and pragmatic adequacy. Some of those criteria are differentiated for grand theories and middle-range theories but are not differentiated by type of data qualitative or quantitative-used to develop the theory. Parse's criteria are structure and process. Structure encompasses historical evolution, foundational elements, and relational statements. Process encompasses correspondence, coherence, and pragmatics. Parse's criteria are appropriate for the critical appraisal of all frameworks and theories, regardless of level of abstraction. Parse also presents a comparison of her own and Fawcett's criteria. PMID- 15802746 TI - Partnering of researcher and practicing nurses for transformative nursing. AB - The purpose of this praxis research was to address the process of a researcher's partnering with practicing nurses to incorporate Newman's theory of health as expanding consciousness as a guide for nursing practice. Initially the nurses were bound by their commitment to help clients within a philosophy of cure, but as they began to engage in a caring relationship of pattern recognition with clients, they experienced the clients' transformations in a way that was transformative for them and that pervaded the total realm of their nursing practice. The process illustrates the immediate applicability of Newman's theory. PMID- 15802747 TI - Testing a middle-range theoretical model of adaptation to chronic pain. AB - The purpose of this cross-sectional, correlational study was to test a middle range theory of adaptation to chronic pain that was deduced from the Roy adaptation model using structural equation modeling. The sample consisted of 200 community-dwelling older adults with a mean age of 76. The Roy adaptation model theoretical framework provided an adequate fit to the data and supported the majority of the hypothesized relationships and findings from prior research. Significant direct effects, however, were found between both focal and contextual stimuli, suggesting that contextual stimuli need to be considered when developing a plan of care for older adults experiencing chronic pain. PMID- 15802748 TI - A discussion of the concept of spirituality. AB - The purpose of this article is to discuss the concept of spirituality. There is a recognized connection between spirituality and health, and nurse leaders have acknowledged the importance of the spiritual as well as the physical in providing optimal nursing care. Spirituality goes beyond religious or cultural boundaries. Spirituality is characterized by faith, a search for meaning and purpose in life, a sense of connection with others, and a transcendence of self, resulting in a sense of inner peace and well-being. A strong spiritual connection may improve one's sense of satisfaction with life or enable accommodation to disability. PMID- 15802749 TI - Exploring the lived experience of waiting for persons in long-term care. AB - The purpose of this study was to describe the meaning of waiting for persons who reside in long-term care settings. Parse's theory of human becoming provided the nursing perspective and a qualitative descriptive-exploratory design was used. The 45 participants were residents in three different long-term care facilities affiliated with a university. Data were gathered through interviews. Three emergent themes formed the following unified description: The experience of waiting is intensifying ire while diversionary immersions reprieve amid unfolding becalming endurance. The themes are discussed in relation to participants' descriptions, the human becoming theory, and related literature. Recommendations for practice and further research are presented. PMID- 15802750 TI - Exploring being: an international dialogue. AB - Nurses and other health professionals are in a privileged position in that they are called to be with people and their families at some of the most important moments in persons' lives. In order to better prepare nurses for this awesome and challenging responsibility, a session of the 11th International Human Becoming Conference held in Niagara Falls, Ontario, Canada, on November 6, 2004, reflected on famous quotations on Being and related Being to nursing practice. The human becoming school of thought is unique in its willingness to face the important but difficult question, what is Being? PMID- 15802751 TI - Quality of life in women following coronary artery bypass graft surgery. AB - The purpose of this study was to determine the effect of coronary artery bypass graft (CABG) surgery on the quality of life of women. Wilson and Cleary's conceptual model of health-related quality of life, which examines five major outcomes (biological variables, symptom status, functional status, general health perceptions, perceived quality of life), was used. Participants included 61 women who provided information by self-report questionnaires before and three months after surgery. The major findings of the study are that women had significantly improved quality of life (p = .004) due to increased satisfaction with health and functioning (p < .001) at three months following CABG surgery. They experienced less angina (p < .001) and shortness of breath (p = .014), although fatigue was unrelieved for the majority of women. Psychological well being improved after surgery for most women (p < .001), with lower anxiety levels (p < .001), greater levels of well being (p = .021), feelings of health (p < .001) and vitality (p = .023). Women reported less use of emotive coping (p = .043), indicating less emotional distress. Nevertheless, 25% of the sample continued to experience severe psychological distress three months after surgery, indicating the need for continued follow-up. PMID- 15802752 TI - The control of tuberculosis: progress & prospect. PMID- 15802753 TI - Tuberculosis control: critical lessons learnt. PMID- 15802754 TI - Leptospiral membrane proteins--variations on a theme? PMID- 15802755 TI - RNA interference & inhibition of viruses. PMID- 15802756 TI - Comparison of immunoreactive proteins of commonly circulating serogroups of Leptospira in Andaman Islands, India. AB - BACKGROUND & OBJECTIVE: Early diagnosis is the key to the treatment of leptospirosis. For development of rapid diagnostic kits, a thorough knowledge about the nature of the proteins expressed by the pathogen during infection is necessary. The present study was undertaken to understand the nature of immunoreactive proteins from commonly circulating serogroups of Leptospira in the endemic Andaman and Nicobar Islands, India. METHODS: Proteins were extracted from six strains of Leptospira representing five different serogroups following four different preparation methods, viz., whole cell lysis by sonication, detergent solubilization, outer and inner membrane isolations, and were subsequently characterized on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE). Immunoblots were made from the sonicated proteins using hyperimmune rabbit antisera, homologous and heterologous patient sera separately. RESULTS: The 67, 65, 45, 43, 35, 32 and 18 kDa major proteins in the whole cell lysate were common among all the five serogroups of Leptospira. The 67, 41, 35, 32, 28 and 22 kDa were the major outer membrane proteins, while 94, 32, 25 and 18 kDa protein were in inner membrane. Immunoblots with hyperimmune rabbit antisera detected 67, 65, 60, 45, 43, 41 and 32 kDa common proteins from the whole cell lysates of all strains while homologous and heterologous patient sera detected 32 kDa as the major immunoreactive protein in all pathogenic serogroups. This protein reacted against specific LipL32 antisera indicating that this protein was LipL32. INTERPRETATION & CONCLUSION: The circulating serogroups of Leptospira have common nature of expression of proteins during human infection. Among several immunoreactive proteins, three (67, 45 and 32 kDa) were recognized as major antigens by both rabbit hyperimmune sera and patients sera while the 32 kDa protein was recognized as the major immunoreactive protein by homologous and heterologous patient sera. These conserved immunoreactive proteins could be utilized in developing indigenous diagnostic tests for leptospirosis. PMID- 15802757 TI - Characterization of a newly established potato tuber moth (Phthorimaea operculella Zeller) cell line. AB - BACKGROUND & OBJECTIVE: Potato tuber moth (PTM), Phthorimaea operculella Zeller is a widely distributed, devastating pest of potatoes attacking the foliage and infest the tubers in both field and store causing serious economic damage. As application of PTM granulovirus (PTM-GV) has shown significant reduction in damage, attempts were made to develop a new cell line from this insect to grow PTM-GV for use as a biopesticide. METHODS: Approximately 100 mg of insect eggs were collected, surface sterilized and crushed gently in a boiling tube aseptically. The tissues were washed with physiological saline, suspended in growth medium and incubated stationary at 28 degrees C. Morphology of cells was studied after staining with Giemsa. Besides karyological and growth curve studies, PCR amplification was also done for rapid amplified polymorphic DNA pattern. RESULTS: A new cell line from the embryonic tissue of PTM was maintained in Mitsuhashi Maramorosch medium supplemented with 10 per cent foetal bovine serum. It is in the 78th passage level and designated as NIV-PTM-1095. Random amplified polymorphic DNA profile analysis indicated this as a new cell line from potato tuber moth and differed from the profiles of two other lepidopteran cell lines maintained in the laboratory. Three different cell types were observed at the 40th passage level and comprised of epithelial-like cells (77%), fibroblast like cells (20%) and giant cells (3%). The chromosome number varied from 54-176. The cell line had a cell doubling time of approximately 42 h during the logarithmic phase of growth. The cell line did not support the multiplication of any of the baculoviruses used in the study. INTERPRETATION & CONCLUSION: Since the new cell line is found to replicate PTM-GV, it may be useful for the propagation of PTM-GV in large scale. Studies to scale up the production of the GV in the cell line and field trials may lead to its widespread use as an eco friendly biopesticide. PMID- 15802758 TI - Production of a growth dependent metabolite active against dermatophytes by Streptomyces rochei AK 39. AB - BACKGROUND & OBJECTIVE: Dermatophytes responsible for causing dermatophytoses in humans have acquired resistance to certain antimycotic drugs. We isolated naturally occurring actinomycetes with an ability to produce metabolites having antimycotic property. The timecourse of antifungal metabolite production in terms of arbitrary units (AU) under optimum conditions was studied. METHODS: Water and soil samples were collected from various locations. The actinomycetes were isolated on starch casein medium and screened for their antifungal activity against yeasts and molds including dermatophytes. One promising isolate which showed a unique, stable and interesting property of inhibiting only dermatophytes was selected and characterized. Optimization of antifungal metabolite production in terms of AU using Trichphyton rubrum as target was done. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of the culture supernatant from the isolate and that of griseofulvin were determined for all dermatophytes. RESULTS: Of the 218 actinomycete isolates, 14 per cent produced the metabolites having antifungal activity. The selected actinomycete, identified as Streptomyces rochei AK 39 produced metabolite, which was active against only dermatophytes whereas yeasts and other molds were resistant to it. Starch casein medium was found to be good for inducing antifungal activity in the isolate. The maximum antifungal metabolite production (400 AU/ml) was achieved in the late log phase, which remained constant during the stationery phase, and it was extracellular in nature. The MIC and MFC values of the culture supernatant from the isolate against the dermatophytes were within the range 1.25 to 5 and 1.25 to 10 AU/ml respectively. INTERPRETATION & CONCLUSION: The metabolite from Streptomyces rochei AK 39 was produced during late log phase and was active against only dermatophytes with a greater potency than griseofulvin. However, this needs further investigation using purified powdered form of the active component. PMID- 15802759 TI - Immunocomb Chlamydia bivalent assay to study Chlamydia species-specific antibodies in patients with coronary heart disease. AB - BACKGROUND & OBJECTIVE: Serological evidences suggested an association between Chlamydia pneumoniae infection and coronary heart disease (CHD). Efficacy of available serological tests for detection of C. pneumoniae antibody has been debated. The present study was carried-out to assess the efficacy of Immunocomb Chlamydia bivalent IgG assay vis-a-vis micro immunofluorescence (MIF) test in detecting C. pneumoniae and C. trachomatis--specific antibodies in patients with CHD. METHODS: Serum samples collected from clinically confirmed cases of CHD (n=114) were subjected to Immunocomb Chlamydia bivalent assay and the standard MIF test. Antibodies specific to C. pneumoniae and C. trachomatis were detected quantitatively. RESULTS: Though Immunocomb Chlamydia bivalent test yielded 73.7 per cent positivity for C. pneumoniae- specific IgG antibody (compared to 50.8% by MIF), the specificity of Immunocomb was found only 32.14 per cent. Positive and negative predictive values of Immunocomb assay were 54.8 and 60.0 per cent respectively. INTERPRETATION & CONCLUSION: The findings of the present study indicated that though Immunocomb assay was inferior to MIF, it can be used as a method for presumptive serology due to its rapidity and ease of performance. Wherever possible, one or more additional tests should also be performed to increase the specificity of such studies. PMID- 15802760 TI - Determination of systemically & locally induced periodontal defects in rats. AB - BACKGROUND & OBJECTIVE: The role of lathyrogens on bone metabolism is unclear, therefore we undertook this study to observe periodontal and systemic alterations in experimental lathyrism in rat and compare these changes to that observed in the locally induced periodontitis group. METHODS: A total of 45 male Wistar rats were equally divided in the lathyritic group (group 1), ligature-induced periodontitis group (group 2), and healthy controls (group 3). Experimental lathyrism was induced by once daily subcutaneous administration of beta aminoproprionitrile (beta-APN), at a dose of 5 mg/0.4 ml per 100 g of body weight for 40 days. Ligature-induced periodontitis was created by tying silk ligatures on the necks of mandibular molars. After 40 days, blood samples were obtained and the animals were decapitated. Radiographic observations, extraction tests, histologic evaluations were performed, and serum ALP activity and gingival tissue IL-1beta levels were measured. RESULTS: Significant alveolar bone resorption around the mandibular molar teeth (P<0.001); lower extraction force levels (P<0.001); higher numbers of lymphocytes and macrophages (P<0.01) (both in connective tissue and epithelium at the dentogingival junction); decreased ALP activity (P<0.001); and increased gingival tissue IL-1beta levels (P<0.001) were observed in groups 1 and 2, compared to those in group 3. ALP activity was higher in group 1 than in group 2 rats (P<0.05). INTERPRETATION & CONCLUSION: Similar radiographical and histopathological findings and comparable increases in gingival tissue IL-1beta levels both in groups 1 and 2 showed that in addition to resorption of alveolar bone, chronic inflammation of periodontium also occurred both in the lathyritic rats as well as in ligature-induced periodontitis group rats. PMID- 15802761 TI - Sensitivity index of antimicrobial agents as a simple solution for multidrug resistance in Salmonella Typhi. AB - BACKGROUND & OBJECTIVE: Multi drug-resistant (MDR) typhoid in India is an escalating problem. MDR isolates of Salmonella Typhi are on rise and are becoming a challenge for timely and appropriate treatment. Occurrence of per cent sensitive (%S), per cent resistant (%R) and per cent intermediate (%I) isolates may vary geographically and treatment decided on the basis of only one of these three subpopulations may lead to selection of inappropriate drug for treatment and thus treatment failure. Determination of sensitivity index (SI) of antimicrobial agents, instead of %S or %R subpopulations, may give clearer insight regarding selection of appropriate antimicrobial for treatment of typhoid. In present work, the data of sensitivity testing were analysed and interpreted both in terms of SI as well as %S, %I and %R. METHODS: A total of 205 isolates of Salmonella Typhi were collected during June 2000 and August 2002 from a network of five institutes- Lady Hardinge Medical College (LHMC, N=110), Ram Manohar Lohia Hospital (RML, N=14), Majeedia Hospital (MH, N=48), Lal's Pathology Lab (LAL, N=28) and All India Institute of Medical Sciences (AIIMS, N=5) on nutrient agar slopes. Of these, 142 isolates were subjected to phage typing and biotyping at National Salmonella Phage Typing Centre, New Delhi. Five isolates resistant to 3-7 and one isolate susceptible to all of total 12 antimicrobial tested were subjected to plasmid analysis. SI for various antimicrobials was determined as the ratio of %S and %R values derived form %RIS analysis using WHONET5. RESULTS: 18 (8.7%) isolates were susceptible to all tested antimicrobials and 124 (60%) were MDR. Of the 142 isolates, 103 were phage type E1 and biotype I. SI of antimicrobials rather than individual %S or %R or %I population presents a better criterion for interpretation of sensitivity testing data as well as selection of the most appropriate antimicrobial for timely treatment. Presence of 140, 48 and 23 Kb size plasmids in all 5 MDR isolates and none in susceptible isolate was observed. INTERPRETATION & CONCLUSION: Re emergence of chloramphenicol sensitivity in Salmonella typhi was observed in the present study. Interpretation in terms of SI criteria warrants that reintroduction of chloramphenicol at present for treatment of typhoid may rebound resistance. Current empiric therapy used for treatment of typhoid may soon become ineffective. SI being a ratio will not only eliminate geographical variation of %RIS data but also its interpretation. SI can provide guidelines for clinicians in remote areas where facilities for sensitivity testing are not available. PMID- 15802762 TI - Expression of opioid receptor-like 1 (ORL1) & mu opioid receptors in the spinal cord of morphine tolerant mice. AB - BACKGROUND & OBJECTIVE: The mechanism underlying the development of tolerance to morphine is not clearly understood though a number of factors have been implicated. One of the likely factors may be increased activity of anti-opioid peptides like nociceptin (also known as orphanin FQ or N/OFQ). N/OFQ and morphine bind to opioid receptor-like 1 (ORL1) receptor and muopioid receptor respectively. The present work was undertaken to investigate the density of ORL1 and mu (mu) receptor expression in the spinal cord of mice after inducing morphine tolerance. METHODS: Swiss albino mice were injected with either morphine (experimental group, n=15) or saline (control, n=15), twice a day for 9 days. The development of tolerance was noted by the hotplate test. Cryostat sections of the cervical region of spinal cord were labeled with specific ligands to localize ORL1 and mu receptors. The density of receptor expression over laminae I-II of spinal cord was evaluated using image analysis system. RESULTS: The morphine treated mice developed tolerance by day 9 as evident by the hot plate test. Both receptors were selectively expressed at a higher concentration over the superficial laminae (I-II) of the dorsal horn, indicating a role in pain processing. An increased expression of ORL1 receptors was also noted over the gray matter around the central canal. Quantitative analysis showed an increased expression of ORL1 and mu receptors though the increase was not statistically significant. INTERPRETATION & CONCLUSION: The present study showed that both, ORL1 and mu-opioid receptors were expressed in areas of the spinal cord, concerned with transmission of pain signals. The density of these receptors increased in the superficial laminae (I-II) though not significantly from control after morphine tolerance. The increase in ORL1 receptors could oppose the analgesic action of morphine, contributing to tolerance. Further studies need to be done to elucidate the mechanism of morphine tolerance. PMID- 15802763 TI - HBsAg carrier status in urban population of Ahmedabad city. PMID- 15802764 TI - India needs social support system for her women. PMID- 15802765 TI - [WHO Technical Report 921]. AB - WHO Technical Report 921 "Prevention and Management of Osteoporosis" was published in 2003. WHO Scientific Group Meeting on Prevention and Management of Osteoporosis was organized in 1998. The members consisted of 21 scientists from all over the world with 2 experts in WHO. Contents of the book included introduction, pathogenesis of osteoporosis and related fractures, epidemiology and risk factors, diagnosis and assessment, prevention and treatment, socioeconomic aspects, delivery of care and education, summary and recommendations. Osteoporosis is a silent epidemic and destroys QOL in overwhelming populations in the world. Risk factors are different from country to country, but nutritional, genetic, life style and other factors are concerned in the pathogenesis of the disease. Appropriate assessment and diagnosis are essential for the evaluation who should be treated. Many kinds of drugs have been developed for the treatment. Many ways for treating patients with fractures have been proposed. However, the establishment of systems of care is needed with aids of both governmental and non-governmental organizations. Finally education of citizens is very important. PMID- 15802767 TI - [The diagnosis of osteoporosis]. AB - The diagnosis of Osteoporosis on WHO Technical Report of Prevention and Management of Osteoporosis in 2003, in addition to present status in Japan, has been reviewed. Since Technical Report of Assessment of Fracture Risk and Its Application to Screening for Postmenopausal Osteoporosis was published by WHO in 1994, the field of osteoporosis has progressed remarkably. In particular, in regard to methods of measuring bone mass or density, diagnosis, assessment of fracture risk, and assessment of osteoporosis, this Technical Report describes evidences, confirmed from current many data, and provides informations available to osteoporosis practice. This Report also shows future direction and issues of osteoporosis which should be resolved. PMID- 15802766 TI - [Trends in the determination of pharmacological intervention for osteoporosis]. AB - WHO scientific group proposed a new strategy to identify cases for pharmacological intervention by integrating multiple factors of fracture risk. Ten independent risk factors for femoral neck fracture were presented including BMD (bone mineral density), previous fracture, bone chemical markers, and low body weight also their respective relative risk values. After adjusted for population risk, these factors were multiplied to obtain the integrated relative risk values. Then, nomograms were made to represent 10 year-fracture risk values to ages depending on the synthetic risk values. WHO group proposed to pharmacologically intervene when 10-yr risk values of femoral neck fracture exceeds 10-15%. It is a subject in many countries how they evaluate such a case finding strategy for pharmacological intervention. PMID- 15802768 TI - [Epidemiology of osteoporosis]. AB - The risk of osteoporotic fracture is lower in Asia and Africa than Caucasian women in the USA and Europe, but worldwide projections showed that it will probably increase markedly in the future. By 2050,the number of hip fracture increased three to four times higher than that in the present, and over half of all hip fracture will occur in Asia. Up to 50% of the variation in peak bone mass may be determined genetically. Sex hormone deficiency plays a major role in development of osteoporosis. In addition, nutrition, physical activity, cigarette smoking, and alcohol consumption also affects bone mass. Modification of these risk factors is important to prevent osteoporosis and consequent fracture in future generations. PMID- 15802769 TI - [Bisphosphontes]. AB - The profitable effects of bisphosphontes on treatment of osteoporosis have been consistently reported in Japan as in the United States and European countries. Alendronate and risedronate increased bone mineral density and reduced the risk of vertebral fracture. Histomorphomeric analysis and microCT have confirmed that normal bone microarchitecture and mineralization are preserved after 3 or 5 years of treatment. Long-term (7 or 10 years) treatment with bisphosphonates seemed to have a sustained effect on BMD and bone turnover, however, the optimal duration of bisphosphonate treatment is unresolved. PMID- 15802770 TI - [Effects of estrogen and selective estrogen receptor modulators on osteoporosis]. AB - Evidence indicates that estrogen and raloxifene hydrochloride, a selective estrogen receptor modulator, prevent bone loss and reduce the risk of bone fractures in postmenopausal women. However, recent reports indicate that the health risks associated with the use of estrogen, which include breast cancer and coronary heart disease (CHD), exceed the benefits. In contrast, raloxifene is associated with a lower incidence of breast cancer and is not associated with CHD. Therefore, the use of raloxifene is preferable for the prevention and treatment of osteoporosis in postmenopausal patients. PMID- 15802771 TI - [Vitamin D and its derivatives as anti-osteoporotic drugs]. AB - Vitamin D, particularly its active form, has been most widely used in Japan for the treatment of osteoporosis. However, clinical evidence for its efficacy as an anti-osteoporotic drug is scarce in terms of fracture prevention. Recent reports suggest that active vitamin D or its analogs may prevent fracture not only through enhancement of intestinal calcium absorption but also by improving bone quality and/or strength independently of bone mass and by improving neuromuscular function to reduce the number of falls. PMID- 15802772 TI - [Vitamin K2 as a protector of bone health and beyond]. AB - Several lives of evidence indicate a protective effect of vitamin K against osteoporosis. Epidemiological studies showed that low vitamin K intake is associated with the increased risk of osteoporosis. Vitamin K2 (menatetrenone, MK 4) has been clinically used in the treatment of patients with osteoporosis in Japan, Korea and Thailand. Previous studies demonstrated the efficacy of vitamin K2 (45 mg/day) to prevent bone loss and reduce the rate of vertebral fractures, although a large, randomized intervention study is anticipated to provide more detailed evidence. Recently, vitamin K2 has been shown to reduce the progression of hepatocarcinoma. Moreover, it has been proposed that vitamin K may also have beneficial effects to prevent atherogenesis. The clarification of molecular mechanisms by which vitamin K2 exerts these salutary effects deserve further investigations. PMID- 15802773 TI - [Treatment of osteoporosis with parathyroid hormone: evidence and perspective]. AB - Parathyroid hormone (PTH) is a promising bone formation-stimulating agent, and the recent large-scale randomized controlled trial (RCT) revealed that parathyroid hormone increases and reduces lumbar bone mineral density and fracture risk, respectively, more potently than bisphosphonates. Moreover, PTH is also effective for male or glucocorticoid-induced osteoporosis. However, the recent studies showed that the co administration of PTH and bisphosphonate is not good. The further accumulation of evidence about the combined or subsequent therapy is expected. PMID- 15802774 TI - [Management of glucocorticoid-induced osteoporosis]. AB - Osteoporosis is the most frequent adverse effect observed during glucocorticoids therapy. The 2004 edition of the guideline on the management and treatment of glucocorticoid-induced osteoporosis has been proposed by The Japanese Society for Bone and Mineral Research. The subjects were patients with using or planning to use oral glucocorticoids for 3 months or longer. The criterion for starting treatment consisted patients with prior fragility fracture and with new fractures during treatment, patients with less BMD than %YAM80, and patients with using as a dose of 5 mg/day or higher (mean daily dose) as prednisolone equivalent. The Bisphosphonates have been recommended as first-line drugs. Active vitamin D3 and vitamin K2 have been recommended as second-line drugs. PMID- 15802775 TI - [Increase in bone mineral density and its effect on fracture risk]. AB - Over the past 10 years a number of clinical trials have evaluated several agents for treatment and prevention of osteoporosis, and demonstrated that these agents can increase bone mineral density (BMD) and reduce the incidence of fracture. Although the relationship between antifracture efficacy and changes in BMD varies greatly among reports, all of them suggest the presence of antifracture effect that is mediated by factors other than BMD. Fracture risk is associated with bone strength and nonskeletal risk factors such as the propensity to fall. Bone strength is primarily determined by BMD, but bone quality such as bone remodeling, structural and material properties is also an important determinant of bone strength. Some of the agents for osteoporosis treatment might provide the atifracture effect mainly through the improvement of bone quality. The mechanism how the reduction in fracture risk is achieved deserves further studies. PMID- 15802776 TI - [Prevention of hip fractures using hip protectors]. AB - Based on the results of 13 randomized controlled trials, programs for the use of hip protectors in institutionalized elderly people with particularly high risks of hip fractures seem to decrease the incidence of hip fractures. On the other hand, there is little indication for the use of hip protectors by elderly people dwelling independently in their own homes. Compliance greatly influences the effect of hip protectors, and continues to be serious problem. PMID- 15802777 TI - [SERMs: an update for clinicians]. AB - With the successful manipulation by molecular pharmacological technology of the estrogenic properties of selective estrogen receptor modulators (SERMs) that are highly organ- and tissue-specific, SERMs have now become available for the management of osteoporosis, as they did for breast cancer earlier. This presentation focuses on raloxifene (RLX), a second-generation SERM and an agent of interest to clinicians engaged in the management of osteoporosis on a day-to day basis. RLX is a unique agent, in that it exhibits similar but distinct effects on bone metabolism from those of estrogen. RLX also appears to exert milder inhibitory effects on bone resorption than bisphosphonates (BPs), another class of antiresorptive agents, while ample evidence suggests that RLX compares favorably with BPs in preventing a variety of non-traumatic fractures. PMID- 15802778 TI - [Long-term bisphosphonate administration and its effect on bone metabolism]. AB - Since bisphosphonates (BPs) suppress bone turnover, potential concerns regarding bone metabolism during long term BP administration have been raised. Now up to 10 years follow-up of clinical trials with data of bone metabolism are available. There was no adverse effect on bone metabolism with long term BP treatment, and increase in bone mineral density and reduction in fracture risk were retained. PMID- 15802779 TI - [Treatment with bisphosphonates: daily or once-weekly oral administration and intravenous injection]. AB - Oral bisphosphonates are among the most effective therapeutics available to manage osteoporosis, which increase bone mineral density and reduce the incidence of fractures. Although daily administration of bisphosphonates has been well tolerated in most clinical use, some patients develop upper gastrointestinal symptoms. Once-weekly bisphosphonates offers convenience to patients while showing equal efficacy, which may also provide improved compliance and a lower risk of gastrointestinal symptoms. Now, both alendronate and risedronate are available in once-weekly formulations in US and European countries, which have efficacy and tolerability possibly better than the once-daily doses. Under certain circumstances, intermittent treatment with injectable bisphosphonates could be an alternative when compliance is at risk. This type of treatment must be applicable to some Japanese osteoporotic patients in the nearest future. PMID- 15802780 TI - [Active vitamin D metabolite and prevention of falls]. AB - Recently, vitamin D metabolites have been internationally recognized to be effective for osteoporosis. The beneficial trend of the drugs for osteoporosis has been more in the prevention of fracture than in the increase of bone mineral density, and more in the prevention of fractures of four extremities compared to spinal fractures. The reason to reduce the fracture frequency of four extremities is due to increase the muscle power, the decrease of body sway and the reduction of falls by the action of the vitamin D. In the study of Japanese, it was declared that administration of active vitamin D significantly reduced the frequency of femoral neck fracture. PMID- 15802781 TI - [Effects of concurrent therapy for patients with osteoporosis]. AB - In the WHO technical report, vitamin D metabolites have been suggested to have a role as an adjunctive therapy when given with an antiresorptive agent. Beneficial effects on BMD have been reported following the addition of calcitriol to alendronate, etidronate, and HRT, but no data on fracture rates are available. Here we summarize our clinical study on the concurrent therapy and review reports on the concurrent treatments. PMID- 15802782 TI - [Role of nutrition in the treatment of osteoporosis]. AB - Calcium intake was reported to be associated with peak bone mass. Vitamin D insufficiency, which is less severe than deficiency, is prevalent in the elderly and known to cause osteoporosis. Protein malnutrition increases the fracture risk due to decreased bone mineral density and muscle weakness. Other nutrients have also been reported to be associated with osteoporosis. Thus nutritional aspect of osteoporosis should be interpreted from the broader perspectives. Since nutritional status greatly varies from one nation to another, we must add our original evidence in Japan to the report from WHO. PMID- 15802783 TI - [Evaluation of exercise as a preventive therapy for osteoporosis]. AB - Osteoporosis and osteoporotic fractures have become an epidemic in the industrialized world. Osteoporosis, low bone mass, is a silent condition with microarchitectural deterioration of the bone structure leading to decreased bone strength and osteoporotic fractures. Physical activity has been advocated as offering a potential means to increase and maintain bone mineral density. Previous cross-sectional studies showed that there is a strong association between exercise and bone mineral density, especially in athletic individuals. However, there might be a self-selection bias; i.e. individuals with larger muscles and bones are more likely to choose an athletic lifestyle. Although there is a report that physical activity is associated with a reduced risk for hip fracture among older community-dwelling women, the effects of vigorous exercises building bone mass is modest and considerably less than bisphosphonates. The proper evaluation of exercise as a preventative therapy for osteoporosis should focus on prevention of falls or osteoporotic fractures. PMID- 15802784 TI - [Osteoporosis and intake of carbohydrates]. AB - Adequate energy intake including carbohydrates is essential to maintain bone mass. Emaciation along with deficiency in nutrients, such as calcium, vitamin D, and protein is a significant risk factor for bone loss, and should be avoided. However, there is no clinical evidence that shows the direct effects of carbohydrate on bone mass. On the other hand, excessive intake of carbohydrates results in obesity, which causes other metabolic diseases such as diabetes mellitus (DM). Therefore, dietary regimen must be balanced in general, and complications and conditions of individual patients should be taken well into account. In addition, energy intake is a basis for adequate exercise in order to maintain physical activity and ideal body weight, which will further decrease the risk of bone fracture. Some indigestible carbohydrates, such as inulin and oligofructose, are shown to increase the availability of minerals from foods, and thus can be beneficial to bone mass. PMID- 15802785 TI - [Jaws of amphibians and reptiles]. AB - Big jaws of amphibians and reptiles are mainly treated in this article. In amphibians enlarged skulls are for the big jaw in contrast with human's skulls for the brain. For example, famous fossils of Homo diluvii testis are ones of salamanders in fact. In reptiles, mosasaur jaws and teeth and their ecology are introduced for instance. PMID- 15802786 TI - [Case of pseudohypoparathyroidism type 1b diagnosed as having hypocalcemia]. AB - The patient (age 46; female) had hypocalcemia without characteristic physical findings of Albright hereditary osteodystrophy in July, 1998. She had no evidence of tetany. Her blood test revealed elevated serum concentration of parathyroid hormone (PTH) in spite of hypocalcemia and hyperphosphatemia. Her Ellsworth Howard test presented decreased excretion of both phosphate and cAMP in urine after PTH injection compared with normal range of response, indicating end-organ resistance to the actions of PTH. Finally, Ellsworth-Howard test, together with lack of clinical manifestations, resulted in diagnosis of pseudohypoparathyroidism type 1b. PMID- 15802787 TI - Significance of survivin mRNA expression in prognosis of neuroblastoma. AB - Neuroblastoma (NB) is the most common malignant solid tumor in childhood, and among all childhood malignancies is second in prevalence only to leukemia. In NB we need to both make an accurate diagnosis and rapidly analyze the expression of genetic prognostic factors such as MYCN, H-ras, and trkA. Moreover, it has recently become important to analyze the expression of survivin mRNA, a member of the inhibitor of apoptosis protein family. Expression of the survivin gene is related to tumorigenesis and inhibition of apoptosis in some malignant tumors. We investigated its expression by reverse transcription-polymerase chain reaction (RT-PCR) in NB cell lines (SK-N-SH, NB-39, and IMR-32), two normal blood cell samples, and 13 clinical NB tumor samples. All three NB cell lines had high levels of mRNA expression for this gene, but normal blood cells had no expression. We detected expression of survivin mRNA in 7 of the 13 NB tumor samples (54%). Two NB patients were in stage I disease, 6 in stage II, and 5 in stage IV(A). Quantitative analysis by RT-PCR revealed that the ratio between survivin mRNA and human glyceraldehyde-3-phosphate dehydrogenase (h-GAPDH) mRNA was very low in stages I and II (0-0.017). In contrast, in advanced NBs (stage IV(A)) the ratio was much higher (0-0.050). The prognoses of the three patients in the advanced stage who had high ratios of expression were poor. A high level of expression of survivin mRNA indicates a high grade of malignancy, high likelihood of recurrence, and poor prognosis. PMID- 15802788 TI - Control of spermidine and spermine levels in rat tissues by trans-4 methylcyclohexylamine, a spermidine-synthase inhibitor. AB - In rat tissues, a decrease in spermidine, accompanied by an increase in spermine was induced by the oral administration (once daily for either 1 week or 1 month) of trans-4-methylcyclohexylamine (4MCHA), a spermidine synthase inhibitor. This is similar to the changes observed in polyamine content when cell growth is arrested. The body-weight gain of the rats tended to decrease with increasing doses of 4MCHA. A decrease in spermidine, combined with a moderate increase in spermine, was observed dose-dependently in all of the tissues tested, with a relatively fast clearance of 4MCHA. Manipulating the polyamine content of tissues, by daily administration of 100 mumol 4MCHA for 1 week, made it possible to estimate the effects of simultaneously added spermidine or spermine on endogenous polyamine contents. The altered polyamine levels, obtained after daily administration for 1 week, were maintained during the extended 1-month period, with growth-dependent alteration. The results show it is possible to produce experimental rats with a higher spermine:spermidine ratio than control rats to investigate the physiological significance of spermidine downregulation and spermine upregulation in vivo. PMID- 15802789 TI - Synergistic effect of green tea catechins on cell growth and apoptosis induction in gastric carcinoma cells. AB - (-)-Epigallocatechin gallate (EGCG), a major component of green tea catechins, is known to inhibit cell growth and to induce apoptosis in a variety of cultured cells. We examined effects of green tea catechins in cultured cells derived from human gastric carcinoma. The proliferation of four cell lines (MKN-1, MKN-45, MKN 74 and KATO-III) was inhibited with EGCG in a dose-dependent manner. The growth of MKN-45 cells was most efficiently inhibited by the treatment (IC(50): 40 muM EGCG) among the four cell lines, while KATO-III cells were most insensitive (IC(50): 80-150 muM) to the EGCG treatment. In addition, (-)-epicatechin (EC) had a major synergistic effect on the induction of apoptosis in MKN-45 cells treated with EGCG; however it had little effect on the inhibition of cell growth induced by EGCG. To study the molecular mechanisms behind the induction of apoptosis by EGCG, the activity of caspases in MKN-45 cells treated with EGCG was examined. Activity levels of caspases-3, -8 and -9 were elevated in EGCG-treated cells, suggesting that these caspases are involved in the apoptosis induced by EGCG. Furthermore, the synergistic effect of EC with EGCG on the induction of apoptosis was specifically canceled by catalase treatment, suggesting that the synergism involves the extracellular production of reactive oxygen species. PMID- 15802790 TI - cDNA cloning and expression of biologically active platelet activating factor acetylhydrolase (PAF-AH) from bovine mammary gland. AB - Platelet activating factor (PAF)-acetylhydrolase (PAF-AH) is an enzyme that hydrolyzes the acetyl ester at the sn-2 position of PAF, and converts it to the inactive metabolite, lyso PAF. This enzyme is distributed widely in the intracellular as well as the extracellular matrix and is believed to be a defense mechanism that protects the host against the toxic effects of PAF and other biologically active oxidized phospholipids. Purification and expression of cDNA cloning of the intracellular and extracellular types of PAF-AH from several sources from different species have been reported. In this study, the cDNA for PAF-AH was cloned by reverse transcription (RT)-PCR from total RNA of bovine mammary gland. The complete amino acid sequences from the cDNA contains 444 amino acids and was identical to that of the PAF-AH isolated from the bovine spleen cDNA library except for two mismatches of amino acid residues (Thr-247 to Met and Ile-431 to Thr). Recombinant PAF-AH was expressed in HEK 293 cells, which exhibited enzyme activity in the in vitro assay system. Furthermore, recombinant bovine PAF-AH was identified by western blot using human plasma PAF-AH antibody as a monomeric polypeptide with a molecular weight of approximately 43 kDa. This protein can be applied to in vivo models to test its protective role against the deleterious PAF actions. PMID- 15802791 TI - The hyperresponsiveness of W/W(v) mice to oral sensitization is associated with a decrease in TCRgammadelta-T cells. AB - We have already reported that WBB6F1-W/W(v) (W/W(v)) mice, which have mutations in the c-kit gene, are highly susceptible to oral sensitization, and that the proportion of TCRgammadelta-T cells among the intraepithelial lymphocytes (IELs) (gammadelta-IELs) of W/W(v) is much lower than in congenic wild-type (+/+) mice. In this study we examined an inhibitory role of gammadelta-IELs in oral sensitization using two different methods. First, wild-type (+/+) mice were sensitized by oral administration of 1.0 mg ovalbumin (OVA) by gavage every day for 9 weeks after anti-TCRgammadelta antibody treatment 4 times. The treatment resulted in an enhanced OVA-specific IgG1 antibody production, active systemic anaphylaxis (ASA), and Th2-dominant cytokine production. Next, W/W(v) mice whose bone marrow cells were reconstituted from C57BL/6J mice for 5 months were sensitized by oral administration of OVA. The OVA-specific IgG1 antibody titer in the bone marrow-reconstituted W/W(v) mice was neither significantly enhanced, nor ASA was induced. The proportion of gammadelta-IELs in the reconstituted mice was much higher than that in the untreated W/W(v) mice. The above findings suggest that the decrease or increase in number of gammadelta-IELs enhances or decreases oral sensitization respectively. These results show that gammadelta-IELs have an important role in the oral tolerance to food antigens. PMID- 15802792 TI - FTR1335 is a novel synthetic inhibitor of Candida albicans N-myristoyltransferase with fungicidal activity. AB - Inhibitors of the fungal enzyme N-myristoyltransferase (Nmt) reduce fungal growth, as this enzyme is essential for viability. We found that a newly synthesized benzothiazole derivative, (1R,3S)-N-{2-[(cyclopeanthylcarbonyl) amino]-benzothiazol-6-yl}-3-[(2-naphthylmethyl) amino] cyclohexanecarboxamide (FTR1335), preferentially inhibited Candida albicans Nmt (CaNmt) in a dose dependent manner. The 50% inhibitory concentration (IC(50)) for CaNmt was 0.49 nM, which was much lower than the 5400 nM IC(50) for human Nmt (HsNmt1). The mode of CaNmt inhibition was competitive with the substrate peptide and non competitive with myristoyl-CoA. Moreover, FTR1335 showed strong antifungal activity in vitro, and the minimum fungicidal concentration for C. albicans was 0.78 microM. These results indicate that FTR1335 might represent a novel family of Nmt inhibitors with fungicidal activity. PMID- 15802793 TI - Analgesic and antioxidant activity of mangiferin and its derivatives: the structure activity relationship. AB - Mangiferin, 2-beta-D-glucopyranosyl-1,3,6,7-tetrahydroxy-9H-xanthen-9-one, obtained directly from methanolic extracts of Bombax ceiba leaves in substantial amounts demonstrated strong antioxidant activity (EC(50) 5.8+/-0.96 mug/ml or 13.74 muM) using DPPH assay comparable to rutin, commonly used as antioxidant for medical purposes. The acetyl and cinnamoyl derivatives were found to be less active than mangiferin whereas, methyl and 3,6,7-trimethylether tetraacetate derivatives were inactive implying that for antioxidant activity, free hydroxyl groups and catechol moiety are essential. Moreover, mangiferin showed hepatoprotective activity against carbon tetrachloride induced liver injury further supporting the free radical scavenging property in the in vivo system. Additionally, plant extracts and mangiferin failed to exhibit acute anti inflammatory activity whereas, it displayed significant analgesic effect in acetic acid-induced writhing and hot plate tests in mice. Using naloxone, it was revealed that plant extracts induced analgesia was independent of opioid receptor, whereas, mangiferin demonstrated significant interaction with it at peripheral site with a slight contribution at the neuronal level. PMID- 15802794 TI - Avena Rhealba inhibits A23187-stimulated arachidonic acid mobilization, eicosanoid release, and cPLA2 expression in human keratinocytes: potential in cutaneous inflammatory disorders. AB - The aim of the present study was to examine the effects of Avena Rhealba (AR) oatmeal extract on the metabolism of arachidonic acid (AA) and eicosanoids as well as on the expression of cytosolic phospholipase A(2 )(cPLA(2)) in the human keratinocyte cell line HaCaT. For this purpose, we examined the effects of AR on basal and A23187-triggered release of [(3)H]-AA from phospholipids and on the production of [(3)H]-labeled metabolites of the cyclooxygenase (CO) and 5 lipoxygenase (LO) pathways. AR was found to inhibit A23187-triggered [(3)H]-AA mobilization from phospholipids (p<0.05) and production of [(3)H]-labeled metabolites of CO (p<0.05) and LO (p<0.05) pathways. These results suggest AR decreases PLA(2)-dependent mobilization of AA from phospholipids. A closer examination of the effects of AR on prostaglandin 6KF1alpha (6KPGF1alpha), the stable metabolite of prostacyclin, revealed dose-dependent inhibition of this AA metabolite. AR also decreased A23187- and tumor necrosis factor alpha-induced cPLA(2) overexpression, as shown by cPLA(2) immunodetection and mRNA expression. These results demonstrate the high potential of AR in the treatment of inflammatory diseases of the skin. PMID- 15802795 TI - Xanthoangelol D isolated from the roots of Angelica keiskei inhibits endothelin-1 production through the suppression of nuclear factor-kappaB. AB - Transcription factor nuclear factor-kappa B (NF-kappaB) has been demonstrated to be important in regulating various gene expressions such as cytokines, adhesion molecules, and endothelin-1 (ET-1) in vascular endothelial cells. In the present study, we show the effects of xanthoangelol, xanthoangelol D, E, and F, which isolated from the root of Angelica keiskei KOIDZUMI (Umbelliferae), on NF-kappaB activation and ET-1 gene expression in cultured porcine aortic endothelial cells (PAECs). Treatments of xanthoangelol D but not xanthoangelol, xanthoangelol E and F markedly suppressed both of basal and tumor necrosis factor-alpha (TNF-alpha) induced NF-kappaB activation in PAECs. To clarify the mechanism of xanthoangelol D-induced suppression on NF-kappaB activation, we evaluated the effects of xanthoangelol D on phosphorylation and degradation of IkappaBalpha, an inhibitory protein bound to NF-kappaB, and obtained evidence that xanthoangelol D selectively suppresses the phosphorylation of IkappaBalpha rather than the degradation of phosphorylated IkappaBalpha. In addition, xanthoangelol D significantly attenuated basal and TNF-alpha-induced prepro ET-1 mRNA expression in PAECs. These results suggest that xanthoangelol D may be useful for the treatment of various vascular diseases involved NF-kappaB activation. PMID- 15802796 TI - Marine alkaloids (-)-pictamine and (-)-lepadin B block neuronal nicotinic acetylcholine receptors. AB - Ascidians (sea squirts) contain a wealth of alkaloids, but their influence over neuronal nicotinic acetylcholine receptors (nAChRs) has not been evaluated. In this study, we examined the effects of two synthetic compounds, (-)-pictamine, a quinolizidine alkaloid from Clavelina picta, and (-)-lepadin B, a decahydroquinoline alkaloid from Clavelina lepadiformis, on major types of neuronal nicotinic receptors (alpha4beta2 and alpha7) expressed in Xenopus oocytes. We found that these alkaloids are potent blockers at these receptors: acetylcholine-elicited currents through alpha4beta2 and alpha7 receptors were blocked by (-)-pictamine with IC(50) values of 1.5 microM and 1.3 microM, respectively, and by (-)-lepadin B with IC(50) values of 0.9 microM and 0.7 microM, respectively. Interestingly, no recovery was observed after the removal of (-)-pictamine in oocytes expressing alpha4beta2 receptors, whereas the inhibited alpha7 currents quickly recovered after the removal of (-)-pictamine. Since there are few compounds that elicit irreversible blocks of alpha4beta2 receptors, (-)-pictamine will be a novel, valuable tool to remove the alpha4beta2 nAChR action from neuronal activities mediated by these two major types of nAChRs. PMID- 15802797 TI - The age-related degeneration of oligodendrocytes in the hippocampus of the senescence-accelerated mouse (SAM) P8: a quantitative immunohistochemical study. AB - The senescence-accelerated mouse (SAM) is known as a murine model for accelerated aging. The SAMP8 shows age-related deficits of learning and memory at an earlier age than control mice (SAMR1). We investigated the changes in oligodendrocytes in the brain of SAMP8, using immunohistochemistry for myelin basic protein (MBP) and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) as an oligodendrocyte marker. SAMP8 at 10 months old showed a decrease in MBP-immunoreactivity (IR) and CNP-IR in the hippocampal CA1 subfield, compared with SAMR1. There were no significant differences in MBP and CNP old in the cerebral cortex and the optic tract between SAMR1 and SAMP8 at 10 months. Furthermore, we measured the area of MBP-IR in the CA1 subfield of both strains and found that the area of MBP-IR in SAMP8 had decreased progressively with age, compared with SAMR1. These results suggest that age-related degeneration of oligodendrocytes had occurred in the hippocampus of SAMP8. PMID- 15802798 TI - Inhibition of hepatocyte growth factor production in human fibroblasts by ursodeoxycholic acid. AB - Hepatocyte growth factor (HGF) stimulates the proliferation of hepatocytes and biliary epithelial cells and protects hepatocytes from apoptosis induced by various stimuli. In view of HGF induction by interferons, substances used for the treatment of chronic hepatitis C, this study was conducted to determine whether ursodeoxycholic acid (UDCA), which is widely used for the treatment of cholestatic liver diseases, modulates HGF production. UDCA did not induce HGF production in human dermal fibroblasts, but it potently inhibited phorbol-12 myristate-13-acetate (PMA)- and cholera-toxin-induced HGF production without affecting cell viability. The inhibitory effects of UDCA were as potent as those of transforming growth factor-beta1 and dexamethasone. Up-regulations of HGF gene expression induced by PMA and cholera toxin were also inhibited by UDCA. Moreover, UDCA dose-dependently inhibited high constitutive HGF production by MRC 5 cells without decreasing cell viability. Deoxycholate, chenodeoxycholate, taurochenodeoxycholate and glycochenodeoxycholate also inhibited cholera-toxin induced HGF production at non-cytotoxic doses. UDCA, however, had no apparent effect on PMA-induced phosphorylation of mitogen-activated protein kinase, which is crucial for HGF induction by PMA. These results indicate that non-cytotoxic doses of UDCA inhibited constitutive and induced HGF production and suggest that UDCA supplemented with HGF or HGF inducers could have a more potential therapeutic effect. PMID- 15802799 TI - Involvement of M3 muscarinic receptors in ACh-induced increase in membrane associated RhoA of rat bronchial smooth muscle. AB - It is known that RhoA is translocated from cytoplasm to cell membrane in bronchial smooth muscle when activated by acetylcholine (ACh) stimulation. In the present study, the effects of selective muscarinic receptor antagonist methoctramine, AF-DX116 (for M(2)) and 4-diphenylacetoxy N-methylpiperidine (4 DAMP; for M(3)) on the ACh-induced rat bronchial smooth muscle contraction and increase in membrane-associated RhoA were investigated to elucidate the muscarinic receptor subtype participating in these responses. To evaluate ACh induced contraction of bronchial smooth muscle, bronchial ring of rat was prepared, suspended in an organ bath and the tension was measured isometrically. To quantify the ACh-induced increase in membrane-associated RhoA protein, western blot analysis was performed by using homogenates of membrane and cytosolic fractions of the rat bronchi. The muscarinic M(2) and M(3) receptors were detected by using RT-PCR in rat bronchial smooth muscle. Both the ACh-induced smooth muscle contraction and increase in membrane-associated RhoA were markedly inhibited by 4-DAMP, but not by methoctramine or AF-DX116. In conclusion, these results indicated contraction for the first time that the activation of RhoA occurs via M(3) receptor in rat bronchial smooth muscle. PMID- 15802800 TI - DY-9760e, a calmodulin antagonist, reduces brain damage after permanent focal cerebral ischemia in cats. 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), a calmodulin antagonist, provides protection against Ca(2+) overload-associated cytotoxicity and brain injury after cerebral ischemia in rats. In this study, we assessed the effect of DY-9760e on ischemic infarct volume in cats subjected to permanent focal cerebral ischemia. DY-9760e was infused for 6 h, beginning 5 min after occlusion of the middle cerebral artery. The infarct volume was measured at the end of drug infusion. DY-9760e, at the dose of 0.25 but not 0.1 mg/kg/h, significantly reduced cerebral infarct volume without affecting any physiological parameters, and its protective effect was mainly evident in the cerebral cortex, where the penumbra, a salvageable zone, exists. The present study demonstrates that DY-9760e protects against brain injury after focal ischemia in a gyrencephalic animal as well as in the rodents reported previously and suggests its therapeutic value for the treatment of acute stroke. PMID- 15802801 TI - Different effects of desipramine on bufuralol 1''-hydroxylation by rat and human CYP2D enzymes. AB - Inhibitory effects of desipramine (DMI) on rat and human CYP2D enzymes were studied using bufuralol (BF) 1''-hydroxylation as an index. Inhibition was examined under the following two conditions: 1) DMI was co-incubated with BF and NADPH in the reaction mixture containing rat or human liver microsomes or yeast cell microsomes expressing rat CYP2D1, CYP2D2 or human CYP2D6 (co-incubation); 2) DMI was preincubated with NADPH and the same enzyme sources prior to adding the substrate (preincubation). When either rat liver microsomes or recombinant CYP2D2 was employed, the preincubation with DMI (0.3 microM) caused a greater inhibition of BF 1''-hydroxylation than the co-incubation did, whereas BF 1''-hydroxylation by rat CYP2D1 was not markedly affected under the same conditions. The inhibitory effect of DMI on BF 1''-hydroxylation by human liver microsomal fractions or recombinant CYP2D6 was much lower than that on the hydroxylation by rat liver microsomes or CYP2D2. Kinetic studies demonstrated that the inhibition-type changed from competitive for the co-incubation to noncompetitive for the preincubation in the case of CYP2D2, whereas the inhibition-type was competitive for both the co-incubation and the preincubation in the case of CYP2D6. Furthermore, the loss of activity of rat CYP2D2 under the preincubation conditions followed pseudo-first-order kinetics. Binding experiments employing the recombinant enzymes and [(3)H]-DMI revealed that CYP2D2 and CYP2D6 were the only prominent proteins to which considerable radioactive DMI metabolite(s) bound. These results indicate that rat CYP2D2 biotransforms DMI into reactive metabolite(s), which covalently bind to CYP2D2, resulting in inactivation of the enzyme. In contrast, human CYP2D6 may also biotransform DMI into some metabolite(s) that covalently bind to CYP2D6, but that do not inactivate the enzyme. PMID- 15802802 TI - Tempol protects against ischemic acute renal failure by inhibiting renal noradrenaline overflow and endothelin-1 overproduction. AB - The effects of tempol, a superoxide dismutase mimetic, on ischemia/reperfusion induced acute renal failure (ARF), noradrenaline (NA) overflow and endothelin-1 (ET-1) overproduction in rats were examined. Ischemic ARF was induced by occlusion of the left renal artery and vein for 45 min followed by reperfusion, 2 weeks after contralateral nephrectomy. Renal functional parameters such as blood urea nitrogen, plasma creatinine concentration, and fractional excretion of sodium, NA concentrations in renal venous plasma, and renal ET-1 contents were determined. Renal function in ARF rats markedly decreased at 1 d after reperfusion. Pre-ischemic treatment with tempol (10, 100 mg/kg, i.v.) dose dependently attenuated the ischemia/reperfusion-induced renal dysfunction. Histopathological examination of the kidney of ARF rats revealed severe renal damages, such as tubular necrosis, proteinaceous casts in tubuli and medullary congestion, which were also significantly suppressed by the tempol treatment. There was a significant increase in NA concentrations in renal venous plasma after the ischemia/reperfusion, and this increase was markedly suppressed by the treatment with tempol. In addition, tempol treatment significantly attenuated the increment of ET-1 content in the kidney exposed to the ischemia/reperfusion. These findings suggest that tempol improves the post-ischemic renal injury by inhibiting the neural activity of renal sympathetic nerve and ET-1 overproduction. PMID- 15802803 TI - Effective NADH-dependent oxidation of 7beta-hydroxy-delta8-tetrahydrocannabinol to the corresponding ketone by Japanese monkey hepatic microsomes. AB - The NADH-dependent activity by hepatic microsomes of Japanese monkeys for 7-oxo Delta(8)-tetrahydrocannabinol (7-oxo-Delta(8)-THC) formation from 7beta-hydroxy Delta(8)-THC exhibited about 70% of the NADPH-dependent activity (100%) at the substrate concentration of 72.7 microM, although NADPH was an obligatory cofactor for maximal activity. Both NADH- and NADPH-dependent activities were significantly inhibited by the typical P450 inhibitors, such as SKF525-A and metyrapone. Both activities were almost completely inhibited by the NADPH-P450 reductase inhibitor diphenyliodonium chloride. The ratio of NADH- and NADPH dependent activities varied significantly according to the substrate concentration. Interestingly, the NADH-dependent activity was higher than that of NADPH at low substrate concentrations of 13-50 microM. The ratio was also affected by the cofactor concentration. In the reconstituted system of CYP3A8 purified from hepatic microsomes of Japanese monkeys as a major enzyme responsible for the NADPH-dependent oxidation, NADH as well as NADPH could sustain the oxidation of 7beta-hydroxy-Delta(8)-THC to the corresponding ketone. The NADH-dependent oxidation of 7beta-hydroxy-Delta(8)-THC by monkey livers is mainly catalyzed by CYP3A8 as well as the NADPH-dependent oxidation. These results indicate that NADH as a cofactor may be also useful for the oxidation of 7beta-hydroxy-Delta(8)-THC, and that the cofactor requirement for the reaction is varied by the concentrations of substrate and/or cofactor. PMID- 15802804 TI - Inhibitory effect of ginsenoside Rb1 and compound K on NO and prostaglandin E2 biosyntheses of RAW264.7 cells induced by lipopolysaccharide. AB - In this study, the antiinflammatory activities of ginsenoside Rb1, which is a main constituent of the root of Panax ginseng (Araliaceae), and of its metabolite compound K, as produced by human intestinal bacteria, on lipopolysaccharide (LPS) induced RAW264.7 cells were investigated. Compound K potently inhibited the production of NO and prostaglandin E2 in LPS-induced RAW 264.7 cells, with IC(50) values of 0.012 and 0.004 mM, respectively. Compound K also reduced the expression levels of the inducible NO synthase (iNOS) and COX-2 proteins and inhibited the activation of NF-kB, a nuclear transcription factor. Compound K inhibited the NO level produced by iNOS enzyme activity in a cell-free system, but did not inhibit COX-1 and 2 activities. When ginsenoside Rb1 was orally administered to rats, compound K, but not ginsenoside Rb1, were excreted in their urine. These findings suggest that ginsenoside Rb1 can be transformed to compound K by intestinal bacteria, and compound K may be effective against inflammation. PMID- 15802805 TI - Effects of oxypeucedanin on hKv1.5 and action potential duration. AB - A furocoumarin derivative, oxypeucedanin, was purified from Angelica dahurica, and its effects on the human Kv1.5 (hKv1.5) channel and on the cardiac action potential duration (APD), were examined using the patch-clamp technique and the conventional microelectrode technique. Oxypeucedanin inhibited the hKv1.5 current in a concentration-dependent manner, with an IC(50) value of 76 nM, while it had no effect on human eag-related gene (HERG) current. Oxypeucedanin induced an initial fast decline of hKv1.5 current during depolarizations. The inhibition of hKv1.5 channel by oxypeucedanin was voltage-dependent, especially at depolarizing pulses between -40 and 0 mV which corresponds to the voltage range of the channel's opening. Oxypeucedanin also slowed the deactivation time course, resulting in a tail crossover phenomenon. Additionally, oxypeucedanin prolonged the APD of rat atrial and ventricular muscles in a dose-dependent manner. These results suggest that oxypeucedanin is a kind of open-channel blocker of the hKv1.5 channel and it prolongs the APD; therefore, it is an excellent candidate as an antiarrhythmic drug for atrial fibrillation. PMID- 15802806 TI - Inhibition of production of reactive oxygen species and gene expression profile by treatment of ethanol extract of Moutan Cortex Radicis in oxidative stressed PC12 cells. AB - Moutan Cortex Radicis (MCR) is one of the most widely used Oriental medicines. In this study, we assessed the reducing effect of ethanol extract of MCR on hydrogen peroxide-induced reactive oxygen production, the main cause of cell damage or death in PC12 cells. The viability of cells treated with 1 mg/ml of MCR was significantly restored from that of oxidative-stressed PC12 cells. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H(2)DCFDA assay. MCR at 1-0.01 mg/ml concentration inhibited ROS production in oxidative-stressed cells. To identify candidate genes responsible for the anti oxidative effects of MCR on PC12 cells, an oligonucleotide microarray analysis was performed. The result of gene expression profiles showed that 10 genes were up-regulated and 7 were down-regulated in MCR plus hydrogen peroxide treated cells compared with hydrogen peroxide treated cells. Among them, heme oxygenase (HO) and cathechol-O-methyltransferase (COMT) are related to regulation of ROS generation and the others are known to regulate cell survival and progression. Subsequently, we performed real-time RT-PCR to quantify the ROS related gene. MCR treatment increased the expression of HO by 370% and COMT by 280% at the concentration of 1 mg/ml. These findings suggest that MCR inhibits the production of ROS and cytotoxicity by oxidative-stressed PC12 cells through over-expression of HO and COMT. PMID- 15802807 TI - Inhibitory effects of Stephania tetrandra S. Moore on free radical-induced lysis of rat red blood cells. AB - Crude preparations of Stephania tetrandra S. MOORE (ST), a traditional herbal medicine, have been used safely for arthritis and silicosis in China. In this study, we demonstrated that ST in vitro protects red blood cells from 2,2-azo-bis (2-amidinopropane) dihydrochloride (AAPH)-induced hemolysis. The inhibitory effect was dose-dependent at concentrations of 10 to 1000 microg/ml. Moreover, tests were carried out to identify the main ingredient of ST that exerts a scavenging effect on free-radicals. Three representative alkaloids, tetrandrine, fangchinoline, and cyclanoline, isolated from ST, were found to have inhibitory activities against AAPH-induced lysis of red blood cells (RBC). Furthermore, the ingestion of 200 mg of ST extract was associated with a significant increase in free-radical scavenging effect of plasma in rats. These results suggest that ST as antioxidant inhibits AAPH-induced hemolysis of RBC both in vitro and in vivo. PMID- 15802808 TI - Identification of Panax species in the herbal medicine preparations using gradient PCR method. AB - In order to identify the existence of Panax species in herbal medicine preparations, the Ginseng specific marker primer was selected and created based on the sequence of Korean ginseng DNA fragment, 359 bp. The gradient PCR was performed on 40 types of the herbal medicines including the 7 types of Araliaceae that are in the same family with the Panax ginseng using the created Ginseng maker primer. As result, Panax notoginseng (Chinese), Panax japonicus (Japanese) and Panax quinquefolius (American), along with Panax ginseng (Korean) were the only ones amplified. However, in the case of Atractylodes lancea, one of the herbal medicines not categorized as Panax species, the DNA was prominently amplified by the Ginseng marker primer. The sequence of the amplified DNA of Atractylodes lancea was identified, resulting in enabling the differentiation from the Panax species by the Restriction Fragment Length Polymorphisms (RFLP) method. In addition, the results of the gradient PCR performed on the herbal medicine preparations that consists of Panax ginseng showed that 290 bp size of the original DNA fragments of Panax ginseng was amplified on the herbal medicine preparations containing Panax ginseng. Therefore, these results suggest a possibility of creating a new testing method for identifying specific herb medicines using the gradient PCR, a molecular biological method not only on Panax ginseng, but also on other herbal medicines and herbal medicine preparations. PMID- 15802809 TI - Genotyping of thiopurine methyltransferase using pyrosequencing. AB - Thiopurine methyltransferase (TPMT) metabolizes thiopurine drugs which are used in the treatment of leukemia and some autoimmune diseases. Previously, 11 mutant alleles of TPMT gene (TPMT*1S, *2, *3A, *3B, *3C, *3D, *4, *5, *6, *7, and *8) have been reported. These mutant alleles may cause life-threatening toxicity in patients exposed to thiopurine drugs, 6-mercaptopurine and azathioprine. We have developed a rapid and accurate protocol for TPMT genotype determination using Pyrosequencing(TM) technology in 96 Japanese subjects. Five fragments of the TPMT gene (exon 4, 5, 7, 8, 10) were amplified by PCR, and the 10 single-nucleotide polymorphisms (SNPs) for TPMT*1S, *2, *3A, *3B, *3C, *3D, *4, *5, *6, *7, and *8 were sequenced. The results of this pyrosequencing method corresponded exactly with those of the DNA sequencing method using BigDye terminator chemistry. We have demonstrated that typing of 10 SNPs can be performed within 30 min. Pyrosequencing has a wide application in the large-scale identification of individual TPMT genotypes. PMID- 15802810 TI - Unidirectional inversion of ibuprofen in Caco-2 cells: developing a suitable model for presystemic chiral inversion study. AB - Intestinal chiral inversion of ibuprofen is still lacking direct evidence. In a preliminary experiment, ibuprofen was found to undergo inversion in Caco-2 cells. This investigation was thus conducted to determine the characteristics and influence of some biochemical factors on the chiral inversion of ibuprofen in Caco-2 cells. The effects of substrate concentration (2.5-40 microg/ml), cell density (0.5-2 x 10(6) cells/well), content of serum (0-20%), coexistence of S ibuprofen (corresponding doses), sodium azide (10 mM), exogenous Coenzyme A (CoA) (0.1-0.4 mM), and palmitic acid (5-25 microM) on inversion were examined. A stereoselective HPLC method based on the Chromasil-CHI-TBB column was developed for quantitative analysis of the drug in cell culture medium. The inversion ratio (F(i)) and elimination rate constant were calculated as the indexes of inversion extent. Inversion of ibuprofen in Caco-2 cells was found to be both dose and cell density dependent, indicating saturable characteristics. Addition of serum significantly inhibited the inversion, to an extent of 2.7 fold decrease at 20% content. Preexistence of S enantiomer exerted a significant inhibitory effect (p<0.01 for all tests). Sodium azide decreased the inversion ratio from 0.43 to 0.32 (p<0.01). Exogenous CoA and palmitic acid significantly promoted the inversion at all tested doses (p<0.01 for all tests). This research provided strong evidence to the capacity and capability of intestinal chiral inversion. Although long incubation times up to 120 h were required, Caco-2 cells should be a suitable model for chiral inversion research of 2-APAs considering the human resourced and well-defined characteristics from the present study. PMID- 15802811 TI - HPLC analysis and pharmacokinetic characteristics of 11-hydroxyaclacinomycin X (ID-6105), a novel anthracycline, in rats and beagle dogs. AB - We investigated the pharmacokinetic characteristics of 11-hydroxyaclacinomycin X (ID-6105), a novel anthracycline, after intravenous (i.v.) bolus administration in rats and beagle dogs. We developed an HPLC-based method to analyze ID-6105 levels in plasma, bile, urine, feces, and tissue homogenates and validated the method in a pharmacokinetic study. The plasma concentration of ID-6105 decreased to below the quantifiable limit (0.02 microg/ml) at 4 and 8 h after i.v. administration in rats at doses of 2 and 10 mg/kg, respectively (t(1/2,alpha) and t(1/2,beta) of 0.78 and 17.8 min at a dose of 2 mg/kg, 0.91 and 176 min at a dose of 10 mg/kg, respectively). The AUC increased with nonlinear pharmacokinetics following the dosage increase from 2 to 10 mg/kg in rats, while the pharmacokinetics were not significantly altered in beagle dogs following a dosage increase from 0.5 to 2.5 mg/kg. Of the various tissues tested, ID-6105 was mainly distributed in the lung, spleen, kidney, adrenal gland, and liver after i.v. bolus administration. ID-6105 levels in the lung or kidney 2 h after i.v. bolus administration were comparable to the initial plasma concentration. However, the ID-6105 concentrations in various tissues 48 h after i.v. bolus administration became too small to measure. The cumulative amounts of ID-6105 found in the bile 48 h after the administration of 2 and 10 mg/kg were calculated to be 26.7 and 18.5% of the initial dose, respectively. The corresponding values in the urine 72 h after i.v. administration were 4.33 and 3.07% of the initial dose, suggesting that ID-6105 is mostly excreted in the bile. In conclusion, our observations indicate that ID-6105 was rapidly cleared from the blood and transferred to tissues such as the lung, spleen, kidney, and liver 2 h after i.v. bolus administration. Moreover, the majority of ID-6105 appears to be excreted in the bile by 24 h after i.v. bolus administration. PMID- 15802812 TI - Synergistic antitumor effect of polyphenolic components of water soluble derivative of propolis against Ehrlich ascites tumour. AB - Effect of two preparation (Croatian and Brazilian) of water-soluble derivative of propolis (WSDP), caffeic acid, quercetin, chrysin, naringenin (components present in WSDP) on the development of Ehrlich ascites tumour (EAT) was evaluated. Test components (50 mg/kg) were given perorally or intraperitoneally 2 h prior the intraperitonel injection of EAT (2 x 10(6)) cells. It was observed that all test compounds effectively inhibited tumour growth and the proliferation of EAT. The volume of ascitic fluid induced by EAT cells and total number of cells present in the peritoneal cavity was markedly reduced in EAT-bearing mice treated with test components. In treated mice the number of polymorphonuclear (PMN) cells in the peritoneal cavity was increased while the number of macrophages was decreased. The macrophage spreading activity revealed that WSDP and all test compounds affected the functional state of macrophages increasing their tumorcidal activity; the effect of WSDP was most pronounced indicating synergistic effect of components present in WSDP. Antitumor activity of WSDP may be the result of different specific mechanism(s) of flavonoids present as compared to individual flavonoid given alone. It is likely that the part of antitumor efficacy of test components against EAT cells was the results of increased activity of macrophages. PMID- 15802813 TI - Increased gene expression by cationic liposomes (TFL-3) in lung metastases following intravenous injection. AB - We recently showed that size, not surface charge, is a major determinant of the in vitro lipofection efficiency of pDNA/TFL-3 complex (lipoplex), even in the presence of serum. In this study, the effect of lipoplex size as a result of interaction with serum proteins on in vitro lipofection and the relationship of this with in vivo lipofection was examined in a murine lung metastasis model. As previously described, the pDNA to lipid ratio (P/L ratio) affected both the size and zeta potential of the lipoplex. In vitro studies also indicated that transgene expression in B16BL6 cells was largely dependent on the size of the lipoplex, both in the absence or presence (50% (v/v)) of serum. An in vivo lipofection experiment showed that predominant gene expression in lungs occurred only in tumor-bearing mice, not in normal mice. Based on the in vitro study, this tumor-related gene expression was not related to lipoplex size in the presence of serum (50% (v/v)), suggesting that the size alteration, as the result of interactions with serum proteins in the blood stream may not play an important role in the case of systemic injections. In addition, the efficient gene expression in tumor-bearing lung was not related to the progression of lung metastases. The area-specific gene expression in tumor-bearing lungs, which was largely dependent on the P/L ratio of the lipoplexes, was observed by fluorescent microscopy. Although the underlying mechanism for the area-specific transgene expression is not clear, it may be related to the interaction of lipoplexes with tumor cells, vascular endothelial cells under angiogenesis and normal cells in the lungs. The possibility that TFL-3 is a useful utility to the targeted delivery of pDNA to lungs and tumor-related lipofection is demonstrated. This result suggests that area-specific gene expression in lung metastases may be achieved by controlling the physicochemical properties of the lipoplex, i.e. the P/L ratio. PMID- 15802814 TI - Expression of multidrug resistance proteins and accumulation of cisplatin in human non-small cell lung cancer cells. AB - In order to understand and overcome multidrug resistance (MDR) of human non-small cell lung cancer (NSCLC), mRNA and protein expression levels of P-glycoprotein (MDR1), multidrug resistance-associated protein 1 (MRP1), and lung resistance related protein (LRP) were investigated and compared with the chemosensitivity and the intracellular/intranuclear cisplatin accumulation of three NSCLC cell lines (Ma-10, Ma-31, and Ma-46). Ma-31 was more resistant than Ma-10 and Ma-46 to cisplatin, carboplatin, etoposide, and paclitaxel. The mRNA level of MDR1 was extremely low, and MDR1 protein was not detected in all cell lines. MRP1 mRNA expression was highest in Ma-31 and lowest in Ma-10, but there was no notable difference between the MRP1 protein expression in three cell lines. LRP mRNA/protein was equally expressed in Ma-10 and Ma-31, but was nominal in Ma-46. The intracellular/intranuclear cisplatin accumulation of the cells was determined to be Ma-31>Ma-46>Ma-10. Thus, MDR1, MRP1, and LRP mRNA and protein expression levels were not correlated with the chemosensitivity or the intracellular/intranuclear cisplatin accumulation of each cell line. The present results indicate that MDR proteins (MDR1, MRP1, and LRP) may not play an important role in the chemoresistance and drug efflux of NSCLC cells. PMID- 15802815 TI - In vitro and in vivo synergism between tetracycline and the cardiovascular agent oxyfedrine HCl against common bacterial strains. AB - The cardiovascular drug oxyfedrine HCl revealed noteworthy in vitro antibacterial action against 501 strains of Gram positive and Gram negative bacteria. It also offered significant protection to mice challenged with a mouse-virulent bacterial strain. Prompted by such results, the present study was carried out to ascertain whether this drug could augment the efficiency of an antibiotic when used in combination with it. For this purpose, ten bacterial strains were selected, which were sensitive to oxyfedrine as well as to six antibiotics, like benzyl penicillin, chloramphenicol, ciprofloxacin, erythromycin, streptomycin and tetracycline. Distinct and statistically significant (p<0.01) synergism was observed between oxyfedrine and tetracycline by disc diffusion tests, compared with their individual effects. The fractional inhibitory concentration (FIC) index of this combination, evaluated by checkerboard analysis, was 0.37, which confirmed synergism between the pair. This synergistic drug duo was further dispensed to infected mice. The results of the mouse-protection tests advocated that the combination was significantly synergistic (p<0.0001), according to Student's 't' test. Hence, the capacity of extended antibiotic therapy in several microbial diseases may be improved with the help of this synergistic drug pair, and the study might throw light on newer directions to contest drug-resistant bacterial infections. PMID- 15802816 TI - Characterization of a computerized assay for rapid and easy determination of leukocyte adhesion to endothelial cells. AB - We report on a facile and rapid computerized in-vitro assay for the quantification of leukocyte adhesion to endothelial cells under static conditions using bovine polymorphonuclear neutrophils (PMN) or human leukaemic Mono Mac 6 cells (MM6) and bovine aorta endothelial cells (BAEC). Images of leukocytes adherent to BAEC monolayers grown in microtiter plates were obtained by a digital camera attached to a conventional microscope and transferred to the public domain NIH ImageJ program for analysis. Using individually adapted program routines adherent leukocytes are easily discriminated and reproducibly quantified. The results obtained with our assay correspond to previous findings and demonstrate the suitability of the described procedure, which can easily be adapted to further standards as proven by the use of two different leukocyte species. This assay lends itself to the screening of pharmacological substances with different mechanism of action that might act on either leukocytes or endothelial cells. PMID- 15802817 TI - Rapid analysis method for paraquat and diquat in the serum using ion-pair high performance liquid chromatography. AB - In the present study, by using IPCC-MS3 (GL Sciences Inc. Tokyo, Japan) as the counter-ion in the mobile phase, we established a simple, quick method of analysis that separated and quantified paraquat and diquat on an ODS column by introducing the deproteinized serum sample directly into HPLC. The calibration curve of paraquat and diquat detected at UV 290 nm showed good linearity when the concentration of the injected sample was in the range 0.1-10.0 microg/ml. The detection limit was 0.05 microg/ml, and the mean recoveries (n=5) added 1.0 microg/ml each of paraquat and diquat to standard serum were 87.5% and 89.1%, respectively, while the RSD were 4.52% and 3.85%. All of these were good results, and the time taken for one analysis was less than 30 min. As a result of employing this analytical method for the analyses in four cases of acute poisoning, it was possible to decide promptly on treatment approaches for all of the present cases. PMID- 15802818 TI - Hypoglycemic and antihyperglycemic effect of Gmelina asiatica LINN. in normal and in alloxan induced diabetic rats. AB - The hypoglycemic and antihyperglycemic effect of alcoholic extract of root of Gmelina asiatica LINN. (G. asiatica) was investigated in normal and in alloxan induced diabetic rats. The blood glucose levels were measured at 0 h and 1, 2, 4, 6, 8 and 16 h after the treatment. The alcoholic extract of G. asiatica showed significant (p<0.05) dose dependent percentage blood glucose reduction in normal (25.8% at 100 mg/kg, 28.9% at 250 mg/kg and 32.4% at 500 mg/kg body weight) and in diabetic rats (26.6% at 100 mg/kg, 32.1% at 250 mg/kg and 48.2% at 500 mg/kg body weight) respectively at 6 h. The antihyperglycemic effect of G. asiatica was compared with the reference standard drug tolbutamide (40 mg/kg). PMID- 15802819 TI - Inhibitory effects of the 5-HT(1A) receptor agonist buspirone on stress-induced hyperglycemia in mice: involvement of insulin and a buspirone metabolite, 1-(2 pyrimidinyl)piperazine (1-PP). AB - Effects of serotonergic anxiolytic buspirone on immobilization-induced hyperglycemia were studied in mice. Stress elicited hyperglycemia in mice. Pretreatment with buspirone significantly reduced immobilization-induced hyperglycemia. Buspirone increased serum insulin levels in both non- and stressed mice. The major metabolite of buspirone, 1-(2-pyrimidinyl)piperazine (1-PP) also increased and this further inhibited immobilization-induced hyperglycemia, since 1-PP increased serum insulin levels in both non-stressed and stressed mice, similar to the increases induced by buspirone. These results suggest that buspirone can reduce stress-induced hyperglycemia by facilitating insulin release. Moreover, 1-PP, a metabolite of buspirone may participate in the effects of buspirone. Since 1-PP is an antagonist of alpha(2) receptors, alpha(2) receptors may be related to effects of 1-PP. PMID- 15802820 TI - A novel immunomodulator, FTY720, prevents development of experimental autoimmune myasthenia gravis in C57BL/6 mice. AB - Prophylactic oral administration of a novel immunomodulator (immunosuppressant), FTY720 (1 mg/kg, three times a week), completely prevented the development of experimental autoimmune myasthenia gravis (EAMG) in C57BL/6 mice. EAMG has been used as an animal model for human myasthenia gravis, and was established by immunizing the mice with acetylcholine receptor (AChR) from Torpedo californica. FTY720 also suppressed the production of both anti-Torpedo californica AChR antibody and anti-mouse AChR autoantibody by the mice, which were observed in mice in which EAMG had become established. These results strongly suggest that FTY720 is a promising candidate for treatment of human myasthenia gravis. PMID- 15802821 TI - Growth suppressing effect of garlic compound diallyl disulfide on prostate cancer cell line (PC-3) in vitro. AB - Prostate cancer is the most predominant cancer in men and prostate cancer related death increases every year. Till date, there is no effective therapy other than androgen ablation therapy. At this stage, induction of apoptosis is considered as a better strategy to control cancer. Previous studies reported that aged garlic extract suppresses cancer growth and enhances immune system against cancer. In the present study, diallyl disulfide, oil soluble organosulfur compound of garlic, was studied for its antiproliferative effect on prostate cancer cells in vitro. The suppression of cell growth was demonstrated by [(3)H]thymidine incorporation assay. Induction of DNA damage was assessed by agarose gel electrophoresis. The results showed that diallyl disulfide inhibited the growth of prostate cancer cells in a dose dependent manner, compared to the control. At 50 microM and 100 microM concentrations, diallyl disulfide induced DNA damage in PC-3 cells. It is concluded that diallyl disulfide, component of aged garlic extract, inhibits proliferation of prostate cancer cells through the induction of apoptosis. PMID- 15802822 TI - The effects of Choto-san on the mRNA expression of Alzheimer's disease related factors in the permanent ischemic rat brain. AB - Choto-san is a Kampo medicines that has been used clinically for the treatment of dementia. We measured the mRNA expressions of some factors related to Alzheimer's disease in a dementia model rat brain. The expressions of beta-amyloid precursor protein, gamma-secretase, alpha7 nicotinic acetylcholine receptor, neprilysin, and insulin degrading enzyme (IDE) were significantly increased on day 4 after permanent occlusion of the bilateral common carotid arteries (2VO). Choto-san inhibited the enhancement of IDE expression caused by 2VO, although it failed to show any effects on the expressions of the other molecules. These results suggest that Choto-san may produce a state in which it is not necessary to induce IDE expression to demonstrate the anti-dementia effects. PMID- 15802823 TI - A pharmacokinetic study of intramuscular administration of bulleyaconitine A in healthy volunteers. AB - The pharmacokinetics of bulleyaconitine A (BLA) after a single dose of 0.2 mg intramuscular injection was evaluated in healthy volunteers. Physical exam, vital signs, clinical laboratory tests and electrocardiogram measurements were monitored to assess the safety and tolerance of the drug. The plasma levels of BLA in serial samples, collected over 15 h, were measured by a validated high performance liquid chromatography (HPLC)-electrospray ionization tandem mass spectrometry (MS-MS) method. It was demonstrated that BLA was absorbed rapidly after intramuscular injection. The pharmacokinetic parameters were as follows: the t(max) value was 0.90+/-0.68 h, the C(max) value was 1.13+/-0.76 ng/ml, the AUC(0-t) was 5.16+/-2.05 ng.h/ml, and t(1/2) was found to be 4.88+/-0.97 h. No subject showed any drug-related clinically significant changes on physical examination, vital signs or laboratory tests. Eight of ten subjects reported a distinct feeling of pain at the site of injection starting approximately at the time of their peak plasma concentration and lasting for 2-6 h. The pain was tolerable, and no subject required additional treatment. PMID- 15802824 TI - Effects of gentianine on the production of pro-inflammatory cytokines in male Sprague-Dawley rats treated with lipopolysaccharide (LPS). AB - This study was undertaken to elucidate the mechanism of anti-inflammatory action of gentianine, a constituent of Gentiana Macrophylla. The effects of gentianine on lipopolysacharide (LPS)-induced production of pro-inflammatory cytokines were investigated in male Sprague-Dawley rats. For the first time, we found that oral administration of gentianine (10-100 mg/kg) suppressed the increases in tumor necrosis factor-alpha (TNF-alpha) (ED(50), 37.7 mg/kg) and interleukin (IL)-6 (ED(50), 38.5 mg/kg) in the sera from the rats challenged with bacterial LPS (100 microg/kg; i.p.). However, LPS induced production of other interleukins, such as IL-alpha, was not significantly altered by gentianine. These results suggest that the potential anti-inflammatory action of gentianine might be at least partly based on the suppressed production of TNF-alpha and IL-6. PMID- 15802825 TI - The endothelium-dependent vasodilator action of a new beverage made of red wine vinegar and grape juice. AB - A new non-alcoholic beverage made of red wine vinegar and grape juice (Budo-no megumi) has been recently demonstrated to lower the blood pressure of human as well as rats. In this study, we pharmacologically analyzed the mechanism of its hypotensive action. The thoracic aorta with intact endothelium was isolated from Sprague-Dawley rats, and incubated with a Tyrode's solution. The beverage in concentrations of 0.25 to 2% relaxed the pre-contracted aorta with an alpha adrenoceptor agonist phenylephrine in a concentration-dependent manner, 2% of which induced 59% relaxation. In contrast, the vasodilator response disappeared in the aorta without endothelium. L-Nitro-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase, significantly reduced the vasodilator effect of the beverage, whereas indomethacin, an inhibitor of cyclooxygenase, hardly affected it. These results suggest that the beverage can activate the nitric oxide synthase activity to exert vasodilation, which may partly explain its previously reported hypotensive action. PMID- 15802826 TI - Activation of caspase-3 in HL-60 cells treated with pyrithione and zinc. AB - The transition metal zinc (Zn) is an endogenous regulator of apoptosis. The ability of Zn to modulate apoptosis is believed to be mediated by the regulation of caspase activity. Previously, we reported that an acute influx of labile Zn induced apoptosis via activation of caspase in human leukemia HL-60 cells treated with a Zn ionophore (Py, pyrithione) and Zn at 1 and 25 microM, respectively. In the present study, we investigated the involvement of caspase-3 in Py (1 microM)/Zn (25 microM)-induced apoptosis in HL-60 cells. Pro-caspase-3 is an inactive form of caspase-3. The processing of pro-caspase-3, a sign of caspase-3 activation, occurred 6 h after treatment with Py/Zn. Proteolysis of poly (ADP ribose) polymerase (PARP), a substrate of caspase-3, was also observed 6 h after treatment with Py/Zn. We also confirmed the elevation of caspase-3 activity as an index of the cleavage of amino acid sequences recognized by activated caspase-3. An inhibitor of caspase-3 attenuated the appearance of the DNA ladder. Taken together, these results indicate that the activation of caspase-3 is partly responsible for the induction of apoptosis in Py/Zn-treated HL-60 cells. PMID- 15802828 TI - Study of the absorption of myo-inositol hexakisphosphate (InsP6) through the skin. AB - Recently, some properties of myo-inositol hexakisphosphate (InsP(6)) are related to its dermatological use as discolouring agent, on preventing calcinosis cutis or due to its important role on premature aging. Some studies also seem to demonstrate a capacity of InsP(6) to inhibit skin cancer. In this paper, a first study of the absorption of InsP(6) through the skin is developed. Due to the correlation between InsP(6) absorption and its urinary excretion, these last values were used to evaluate this process. It was found that using a moisturizing cream as vehicle, the InsP(6) sodium salt was absorbed at significantly higher amounts than the InsP(6) calcium-magnesium salt. Maximum InsP(6) urinary concentrations were observed approximately at 14 d of 2% InsP(6) topical cream application, and gave 66.35+/-5.49 mg/l urinary InsP(6) when the sodium salt was used and 16.02+/-2.61 mg/l urinary InsP(6) when the calcium-magnesium salt was applied. When the InsP(6) topical cream administration ceased, the InsP(6) urinary excretion fell dramatically approximately during a period of 10 d. From these results, it can be deduced that by topical administration InsP(6) can achieve important concentrations in tissues and biological fluids, this demonstrating that it is possible to propose the topic use as a new InsP(6) administration route. PMID- 15802827 TI - Limited role of P-glycoprotein in the intestinal absorption of cyclosporin A. AB - The contribution of P-glycoprotein (P-gp) to the intestinal absorption of cyclosporin A (CsA) was investigated by comparing the in vivo pharmacokinetics of CsA in P-gp knockout mice versus wild-type mice following both oral and intravenous administration and by examining the transport of CsA across Caco-2 cell monolayers. The apparent oral bioavailability of CsA in P-gp knockout mice was 1.55-fold larger than in wild-type mice, leading to an apparent absolute bioavailability of 41.8%. A concentration dependent efflux transport of CsA across Caco-2 cell monolayers was found, which exhibited saturation at a CsA concentration of 1 muM. These results suggest that the involvement of P-gp in the intestinal absorption of CsA is not as profound as was previously thought. PMID- 15802829 TI - Nardostachys chinensis glycoside induces characteristics of neuronal differentiation in rat pheochromocytoma PC12 cells. AB - Rat pheochromocytoma PC12 cells undergo neuronal differentiation in response to nerve growth factor. We show here that exposure of PC12 cells to Nardostachys chinensis glycoside induces the outgrowth of neurites, increases the activity of AChE, triggers cell cycle arrest in G1 and enhances the expression of growth associated protein 43 (GAP-43). Both the outgrowth of neurites and the increase in AChE activity are prevented partly by PD98059, a specific inhibitor of MEK1. These results suggest that N. chinensis glycoside induces the characteristics of neuronal differentiation in PC12 cells via the mitogen-activated protein kinase (MAPK)-related signal cascade. PMID- 15802830 TI - Scale-up of high shear granulation based on the internal stress measurement. AB - Scale-up of wet granulation in a vertical high shear mixer was conducted. Pharmaceutical excipient powders composed of lactose, cornstarch and micro crystallinecellulose, and hydroxypropylcellulose as a binder were mixed together and then granulated with purified water under various operating conditions and vessel scales. A novel internal stress measurement system was developed and stress of normal and tangential directions that granules received from the agitator blade during the granulation was continuously measured. The results indicated that granules received stress mainly from the tangential direction, which also showed the largest value near at the vessel wall. The effects of the agitator tip speed and the centrifugal acceleration on the measured stress was investigated. It was found that the tip speed of the agitator blade could be the main factor for the granule growth. The physical properties such as strength, size distribution and compressibility of granules prepared by changing the operating conditions and the vessel scales were evaluated and the scale-up characteristics of high shear granulation were investigated experimentally. The results showed that these physical properties had linear correlations with the tip speed. It was finally concluded that the scale-up of high shear granulation could be well conducted by means of the tip speed of the agitator blade. PMID- 15802831 TI - Stereoselective synthesis of beta-hydroxyphenylalanines using imino 1,2-Wittig rearrangement of hydroximates. AB - The imino 1,2-Wittig rearrangement of hydroximates containing a furan ring provides a novel method for the synthesis of beta-hydroxy-alpha-amino acids. Upon treatment with LDA, hydroximates smoothly underwent the rearrangement to give Z-2 hydroxyoxime ethers in good yield, which were converted into both cis- and trans oxazolidinones with high stereoselectivity. The cis- and trans-oxazolidinones were stereoselectively converted into erythro- and threo-beta hydroxyphenylalanines, respectively, via the oxidative cleavage of a furan ring, ring-opening of oxazolidinone, and deprotection. PMID- 15802832 TI - Biotransformation of benzaldehyde-type and acetophenone-type derivatives by Pharbitis nil hairy roots. AB - The glucosylation of some coumarin and flavone derivatives on incubation with the hairy roots of morning glory (Pharbitis nil) was previously reported. We further studied the biotransformation of benzaldehyde- and acetophenone-type derivatives. Vanillin and isovanillin were reduced to alcoholic derivatives and glucosylated at the phenolic and the alcoholic hydroxyl groups. In the case of 3,4 dihydroxybenzaldehyde, the formyl group was reduced and the 3-hydroxyl or 4 hydroxyl groups were glucosylated to give monoglucosides. The 3-hydroxyl group was predominantly glucosylated to the 4-hydroxyl group. 4-beta-D Glucopyranosyloxy-3-methoxybenzylalcohol was obtained in low yield. In time course experiments with vanillin, it was found that the high-level reduction of the formyl group and glucosylation of the phenolic hydroxyl group occurred, and finally 4-O-beta-D-glucopyranosylvanillylalcohol was obtained as the main product. In the case of 3,4-dimethoxybenzaldehyde, 3,4,5-trimethoxybenzaldehyde, and salicylaldehyde, the formyl groups were reduced, and then the hydroxyl groups at the benyl position were glucosylated to give alcoholic glucosides in relatively high yields. In 4-hydroxy-3-methoxyacetophenone, the 4-hydroxyl group was glucosylated and two dimerized glucosides, biphenyl and biphenylether types, were obtained in low yields. In acetophenone, 1-beta-D-glucopyranosyloxy-1 phenylethane and 2-beta-D-glucopyranosyloxyacetophenone were obtained. As mentioned above P. nil hairy roots showed various biotransformative activities including glucosylation of phenolic and benzylic hydroxyl groups, reduction of the formyl group near the benzene ring, and phenol oxidation dimerization. The glucosylation reaction was especially interesting for the production of valuable glucosides. PMID- 15802833 TI - X-ray structure characterization of palladium(II) ternary complexes of pyridinedicarboxylic and phthalic acid with phenanthroline and bipyridine. AB - The crystal structures of the series of four ternary complexes, [Pd(phen)(2,6 PDCA)].4H(2)O (1) (phen=1,10-phenanthroline; 2,6-PDCA=2,6-pyridinedicarboxylic acid), [Pd(bpy)(2,3-PDCA)].3H(2)O (2) (bpy=2,2'-bipyridineand; 2,3-PDCA=2,3 pyridinedicarboxylic acid) and [Pd(phen)(PHT)].2.5H(2)O (3) (PHT=o-phthalic acid ) and [Pd(bpy)(PHT)].1.5H(2)O (4), are determined and the coordination modes of palladium(II) ternary complexes are characterized. All complexes take the mononuclear Pd(II) complexes, in which central Pd(II) atom of each complex has a similar distorted square-planar four coordination geometry. In all complexes, the aromatic heterocyclic compounds, phen and bpy, behave as a bidentate N, N' ligand. In the complex 1 and 2, 2,6-PDCA and 2,3-PDCA behave as a bidentate N, O ligand, and in complex 3 and 4, PHT behaves as a bidentate O, O' ligand. PMID- 15802834 TI - Application of a stepwise flow ratiometry without phase separation to the determination of the chloroform/water distribution coefficients of volatile diazines. AB - The chloroform/water distribution coefficients (K(D)) of sixteen diazine compounds were determined by a stepwise flow ratiometry. An aqueous solution of analyte was delivered and merged with chloroform. The flow rate ratio of both the phases was varied stepwise under a constant total (chloroform+aqueous) flow rate. The analyte was extracted to chloroform while both the phases, which were segmented by each other, were passing through an extraction coil. The segmented stream was then led to a UV/Vis detector directly without phase-separation. The absorbance of the chloroform and aqueous phases (A(o) and A(a), respectively) was each measured at the maximum absorption wavelength of the analyte. The plots of A(-1) against R(f), (AR(f))(-1) against R(f)(-1), and AR(f) against A gave straight lines, where A was A(o), A(a) or the sum of them (A(S)). The K(D) of the analyte was calculated from the slopes and intercepts of the plots. The log K(D) values obtained for the analytes (-0.5-1.4) were agreed well with the values measured by a shake-flask method. The present method is simple, rapid (5 min/determination) and applicable to the volatile compounds with reasonable precision (standard deviation of log K(D)<0.07). PMID- 15802835 TI - Three new cholinesterase-inhibiting cis-clerodane diterpenoids from Otostegia limbata. AB - Three new tricyclic cis-clerodane type diterpenoids trivially named as limbatolide A (1), limbatolide B (2) and limbatolide C (3) have been isolated from the roots of Otostegia limbata along with two known compounds; oleanic acid and beta-sitosterol. The structure elucidation of the new compounds was based primarily on two-dimensional (2D) NMR techniques. Compounds 1-3 displayed inhibitory potential in a concentration-dependent manner against acetylcholinesterase (AChE; EC 3.1.1.7) and butyrylcholinesterase (BChE; EC 3.1.1.8) enzymes, respectively. PMID- 15802836 TI - Constituents of holothuroidea, 14. Isolation and structure of new glucocerebroside molecular species from the sea cucumber Stichopus japonicus. AB - Five glucocerebroside molecular species, SJC-1-SJC-5, have been isolated from the less polar lipid fraction of a chloroform/methanol extract of the sea cucumber Stichopus japonicus. The structures of these glucocerebroside molecular species were determined on the basis of chemical and spectroscopic evidence. SJC-1, SJC 2, and SJC-3 are typical sphingosine- and phytosphingosine-type glucocerebroside molecular species with nonhydroxylated and hydroxylated fatty acyl moieties. SJC 4 and SJC-5 are also sphingosine-type glucocerebroside molecular species with hydroxylated fatty acyl moieties, although they are new glucocerebroside molecular species with unique sphingosine bases. PMID- 15802837 TI - Inhibitors of nitric oxide production from the flowers of Angelica furcijuga: structures of hyuganosides IV and V. AB - The methanolic extract from the flowers of Angelica furcijuga KITAGAWA was found to inhibit nitric oxide production in lipopolysaccharide-activated mouse peritoneal macrophages. From the methanolic extract, two new glycosides, hyuganosides IV and V, were isolated together with 28 known constituents. The structures of the new constituents were determined on the basis of chemical and physicochemical evidence. Furthermore, the inhibitory effects of 11 coumarin constituents on nitric oxide production were examined. Among them, 3'-angeloyl cis-khellactone (IC(50)=82 microM), (S)-(-)-oxypeucedanin (57 microM), imperatorin (60 microM), isoepoxypteryxin (53 microM), and isopteryxin (8.8 microM) showed inhibitory activity. PMID- 15802838 TI - Synthesis of the new furo[3,2-h]isoquinoline alkaloid, TMC-120B from Aspergillus ustus. AB - A total synthesis of a new furo[3,2-h]isoquinoline alkaloid TMC120-B (2), isolated from Aspergillus ustus together with two related compounds, has been completed in sixteen steps. The key step is the synthesis of the appropriate 3,7,8-trisubstituted isoquinoline framework (23) based on a thermal electrocyclic reaction of the 1-aza 6pi-electron system involving the benzene double bond. In addition, the microwave assisted electrocyclic reaction of this system was newly performed. PMID- 15802839 TI - Hydrogen-bonding abilities for phenols assessed by quantitative analyses of their partition coefficients derived from different partitioning systems. AB - We recently proposed a new hydrogen-accepting scale, S(HA), on the basis of the heat of formation calculated by the conductor-like screening model (COSMO) method. In this work, the same approach was applied to a series of compounds with a common hydrogen-donor group. Thus the S(HA) values for monosubstituted phenols were calculated and used for correlating their log P(oct) values (P(oct): 1 octanol/water partition coefficient) with log P(CL) (P(CL): chloroform/water partition coefficient) and log P(E) (P(E): butyl ether/water partition coefficient). It was demonstrated that the S(HA) parameter works effectively, providing excellent correlations whose physicochemical meanings are well rationalized in terms of hydrogen-bonding characteristics of the substituents. PMID- 15802840 TI - N-Arylpiperazine-1-carboxamide derivatives: a novel series of orally active nonsteroidal androgen receptor antagonists. AB - A novel series of N-arylpiperazine-1-carboxamide derivatives was synthesized and their androgen receptor (AR) antagonist activities and in vivo antiandrogenic properties were evaluated. Reporter assays indicated that trans-2,5 dimethylpiperazine derivatives are potent AR antagonists, and in this series trans-N-4-[4-cyano-3-(trifluoromethyl)phenyl]-N-(2,4-difluorophenyl)-2,5 dimethylpiperazine-1-carboxamide (18 g, YM-175735) exhibited the most potent antiandrogenic activity. Compared to bicalutamide, YM-175735 is an approximately 4-fold stronger AR antagonist and has slightly increased antiandrogenic activity, suggesting that YM-175735 may be useful in the treatment of prostate cancer. PMID- 15802841 TI - Synthesis and biological activities of 4-phenyl-5-pyridyl-1,3-thiazole derivatives as p38 MAP kinase inhibitors. AB - A novel series of 4-phenyl-5-pyridyl-1,3-thiazole analogues possessing potent in vitro inhibitory activity against p38 mitogen-activated protein kinase and the release of tumor necrosis factor-alpha (TNF-alpha) from human monocytic THP-1 cells stimulated by lipopolysaccharide has been identified. Subsequent structure activity relationship (SAR) studies and optimization for absorption, distribution, metabolism, and elimination (ADME) profiles led to the identification of compounds 7 g and 10b as orally active lead candidates that block the in vivo production of proinflammatory cytokine (TNF-alpha). In pharmacokinetic studies, compound 10b showed good oral administration in mice and demonstrated significant in vivo anti-inflammatory activity in an anti-collagen monoclonal antibody-induced arthritis mouse model (minimum effective dose (MED)=30 mg/kg). Further elucidation of this class of compounds may provide novel anti-inflammatory agents, such as anti-rheumatoid arthritis drugs. PMID- 15802842 TI - Flavonoids from Millettia nitida var. hirsutissima. AB - From the stems of Millettia nitida var. hirsutissima, three new isoflavone glycosides, formononetin 7-O-beta-D-(6''-ethylmalonyl)-glucopyranoside (1, hirsutissimiside A), 5-O-methyl genistein 7-O-alpha-L-rhamnopyranosyl-(1-->6) beta-D-glucopyranoside (3, hirsutissimiside B), retusin 7,8-di-O-beta-D glucopyranoside (4, hirsutissimiside C) and two known isoflavone glycosides (2) and (5) have been isolated. The structures of the compounds were determined by spectroscopic and chemical means. PMID- 15802843 TI - Spectrophotometric determination of plasma and red blood cell cholinesterase activity of 53 fruit farm workers pre- and post-exposed chlorpyrifos for one fruit crop. AB - We sought to investigate the early biological effects of chlorpyrifos among 53 Thai fruit farm workers by measuring the plasma cholinesterase (PChE) and red blood cell cholinesterase (AChE) activities, a biomarker of organophosphate (OPs) pesticide during one fruit crop. The ChE activity (V(m)/K(m)) was spectrophotometrically analyzed before and after exposing to chlorpyrifos. The V(m)/K(m) values of both non-spraying and spraying seasons are found as normal distribution pattern. The median PChE and AChE activities among farm workers in the non-spraying season were 2.3 x 10(-3) s(-1) and 7.26 x 10(-5) s(-1), respectively. The median PChE and AChE activities of the farm workers in the spraying season were 2.02 x 10(-3) s(-1) and 5.95 x 10(-5) s(-1), respectively. The mean V(m)/K(m) values of PChE shifted left (t-test, p=0.013), indicating a decrease in PChE activity in the farm workers exposed to chlorpyrifos. However, the V(m)/K(m) values of AChE in nonspraying season and in the spraying season were not different (t-test, p=0.246). We propose that PChE activity can be used as a biomarker for monitoring early toxicity induced by chlorpyrifos insecticide. PMID- 15802844 TI - Catalytic action of triarylstibanes: oxidation of benzoins into benzyls using triarylstibanes under an aerobic condition. AB - Benzoins are simply oxidized to benzils in excellent yields with a catalytic amount of triarylstibanes under an aerobic condition. This catalytic oxidation is heteroatom-specific in the antimony compound and no reaction take place with other group 15 reagents such as triphenylphosphane, -arsane and -bismuthane. The reaction should involve an oxidation-reduction cycle between stibane Sb(III) and stiborane Sb(V) under air. PMID- 15802845 TI - Caesaldecan, a cassane diterpenoid from the leaves of Caesalpinia decapetala. AB - A new cassane diterpenoid, caesaldecan, was isolated from Caesalpinia decapetala with eight known compounds, spathulenol, 4,5-epoxy-8(14)-caryophyllene, squalene, lupeol, trans-resveratrol, quercetin, astragalin, and stigmasterol. The (1)H- and (13)C-NMR spectra of the new compound were completely assigned by using a combination of 2D NMR techniques, namely, (1)H-(1)H COSY, HMQC, HMBC, and ROESY. PMID- 15802846 TI - Preparation of dry powder inhalation with lactose carrier particles surface coated using a Wurster fluidized bed. AB - An attempt was made to produce carrier particles for dry powder inhalation with lactose carrier particles surface-coated using a Wurster fluidized bed. The lactose carrier particles were coated with lactose aqueous solution containing hydroxypropyl methyl cellulose (HPMC) as a binder using a Wurster coating apparatus. Drug/carrier powder mixtures were prepared consisting of micronized salbutamol sulfate and lactose carriers under various particle surface conditions. These powder mixtures were aerosolized by a Jethaler((R)), and the in vitro deposition properties of salbutamol sulfate were evaluated by a twin impinger. The in vitro inhalation properties of the powder mixture prepared using the coated lactose carrier differed significantly compared with those of the powder mixture prepared using the uncoated lactose carrier, indicating improvements in in vitro inhalation properties of sulbutamol sulfate. In vitro inhalation properties increased with the surface coating time. This surface coating system would thus be valuable for increasing the in vitro inhalation properties of dry powder inhalation with lactose carrier particles. PMID- 15802847 TI - 2'-Amino-alpha-chloroacetophenone as a valuable tool for the synthesis of conveniently-substituted alpha,beta-epoxychalcone derivatives. AB - The synthesis of conveniently substituted 2'-amino-alpha,beta-epoxychalcones is described. They were obtained through Darzens condensation of 2'-amino-3',5' dimethoxy-alpha-chloroacetophenone with benzaldehydes. The latter can undergo different cyclization possibilities and afford a variety of flavonoid analogs with biological potential. PMID- 15802848 TI - Discovery of 2-aminothiazole-4-carboxamides, a novel class of muscarinic M(3) selective antagonists, through solution-phase parallel synthesis. AB - Synthesis and structure-activity relationship of a new class of muscarinic M(3) selective antagonists were described. In the course of searching for a muscarinic M(3) antagonist with a structure distinct from those of the 2-(4,4 difluorocyclopentyl)-2-phenylacetamide derivatives, we identified a thiazole-4 carboxamide derivative (1) as a lead compound in our in-house chemical collection. Since this compound (1) showed relatively low binding affinity (K(i)=140 nM) for M(3) receptors in the human binding assays, we tried to improve its potency and selectivity for M(3) over M(1) and M(2) receptors by derivatization of 1 through a combinatorial approach. A solution-phase parallel synthesis effectively contributed to the optimization of each segment of 1. Thus, we have identified a cyclooctenylmethyl derivative (3e) and a cyclononenylmethyl derivative (3f) as representative M(3) selective antagonists in this class. PMID- 15802849 TI - Permeability of ionized salicylate derivatives through guinea pig dorsal skin. AB - pH-dependency of skin permeability to salicylic acid was examined in excised guinea pig dorsal skin. Permeation followed the pH-partition theory at acidic pH. However, above pH 5.0 the observed permeability coefficients were larger than the estimated values obtained from the ratio of the undissociated forms. These findings are quite different from those obtained using the same drug and a silicone rubber membrane, in which permeability coefficients were consistent with the pH-partition theory. The findings suggested that permeation of salicylate as anions occurred at a neutral skin pH. The permeability coefficient of the ionized form was estimated to be about 1.6% of the nonionized form. We also examined the skin permeability of salicylate and its five 5-substituents and two 3 substituents at pH 7.4. We investigated the relationship between their permeability coefficients and the physico-chemical properties of the substituents. Multi regression analysis on the permeability coefficients showed a parabolic relationship between the values of the hydrophobic parameter (pi) and the logarithms of the permeability coefficients. These findings suggested that the ionic permeation pathway of salicylate derivatives is controlled by hydrophobic as well as hydrophilic properties. PMID- 15802850 TI - Design and synthesis of a novel water-soluble NMDA receptor antagonist with a 1,4,7,10-tetraazacyclododecane group. AB - Polyamines, especially spermine, inhibit N-methyl-D-aspartate (NMDA) receptors as open channel blockers. Two types of water-soluble NMDA receptor antagonist, ACCn (1) and TGCn (2), with a 1,4,7,10-tetraazacyclododecane cyclic polyamine group, were synthesized and the effects of both compounds on NMDA receptors were studied using voltage-clamp recordings of recombinant NMDA receptors expressed in Xenopus oocytes. These compounds inhibited macroscopic currents in both NR1/NR2A and NR1/NR2B receptor subtypes in oocytes voltage-clamped at -70 mV. Inhibition by the compounds of NR1/NR2A receptors were more prominent than that of NR1/NR2B receptors. The inhibitory effects of ACCn (1) on both NMDA receptors were more potent than those of TGCn (2). PMID- 15802851 TI - An easy access to 7-methyl-2-naphthalenecarbonitrile. AB - A practical and cost-effective procedure has been developed for the synthesis of 7-methyl-2-naphthalenecarbonitrile, the precursor of the anticoagulant agents YM 60828 or YM-96765. This new route generates the key intermediate in only two steps from readily available 3-cyanopropionaldehyde dimethyl acetal and m tolualdehyde, without requiring chromatographic purification. The synthesis involves condensation of the cyano derivative with the aldehyde and subsequent cyclodehydration. PMID- 15802852 TI - In vitro alpha-glucosidase inhibitory effect of Zn(II) complex with 6-methyl-2 picolinmethylamide. AB - We found alpha-glucosidase inhibitory effect of Zn(II) complex with 6-methyl-2 picolinmethylamide (6mpa-ma) which showed the highest blood glucose lowering effect in Zn(II) complexes with picolinamide derivatives in KK-A(y) mice. The Zn(II) complex showed strong alpha-glucosidase inhibitory activity greater by about eighty times (substrate: maltose) and forty times (substrate: sucrose) compared with acarbose. PMID- 15802853 TI - Myrothenones A and B, cyclopentenone derivatives with tyrosinase inhibitory activity from the marine-derived fungus Myrothecium sp. AB - New 3-amino-5-ethenylcyclopentenones, myrothenones A (4) and B (5), were isolated together with known 6-n-pentyl-alpha-pyrone (1), trichodenone A (2), and cyclonerodiol (3) from the marine algicolous fungus of genus of Myrothecium. The structure and absolute stereochemistry of the new compounds were established by spectral interpretation and X-ray analysis. Compounds 1 and 4 exhibited a tyrosinase inhibitory activity with IC(50) value of 0.8 and 6.6 muM, respectively, which are more active than kojic acid (IC(50), 7.7 muM) currently being used as a functional personal-care compound. PMID- 15802854 TI - Temporal variation of excess mortality rate from solid tumors in mice irradiated at various ages with gamma rays. AB - The influence of age at the time of irradiation on the lifetime risk for excess mortality from solid tumors, and on the temporal pattern of variation in the excess mortality rate, was analyzed using data obtained from a study of female B6C3F1 mice, which was conducted at the National Institute of Radiological Sciences, Chiba, Japan. Mice were irradiated with 1.9-Gy gamma rays at day 17 in intra-uterine age, or day 0, 7, 35, 105 or 365 in postnatal age. Control and irradiated mice were allowed to live out their entire life span under a specific pathogen-free condition. The primary cause of death for each mouse was determined by macroscopic and microscopic examination. The lifetime excess mortality from solid tumors was apparently higher in the mice irradiated during the neonatal to puberty period than in the mice irradiated during the intra-uterine or adult period. The median of time for manifestation of lifetime excess mortality since irradiation was shortest among mice exposed at 365 days of age and longest among mice exposed at 17 days of intra-uterine age. The excess mortality rate at any attained age was not independent of the age at irradiation. The excess mortality rate increased with increasing age, and the excess relative risk decreased with increasing age. The temporal variations of the excess mortality rate and background mortality rate were analyzed using the additive multi-stage model, which includes the assumptions that radiation-related carcinogenesis superimposes on background carcinogenesis, and that both radiation-related and background carcinogenesis involve multiple stages. The results of the analysis strongly suggested that the number of stages for manifestation of radiation-related carcinogenesis was less than that in background carcinogenesis for various types of solid tumors, and that the majority of stages were common in both radiation related and background carcinogenesis. The additive multi-stage model well described the observed findings on the length of the latent period and temporal variations of the excess mortality rate and excess relative risk. It should be stressed that the magnitude of the lifetime risk was not only determined by a decrease in the number of hits for carcinogenesis but was also determined by another parameter which decides the initial value of excess mortality rate. Furthermore, we estimated the rate of decrease in the number of remaining hits for carcinogenesis, and it was found that the rate of decrease in the number of remaining hits was higher in several irradiated groups than that in the background carcinogenesis. However, radiation-induced genomic instability and/or delayed mutation may be of secondary importance when radiation was delivered promptly, because the present analysis revealed that the major action of radiation took place soon after irradiation, as one or more hits for transitions of stages for carcinogenesis. PMID- 15802855 TI - The elevation of p53 protein level and SOD activity in the resident blood of the Misasa radon hot spring district. AB - To clarify the mechanism by which radon hot springs prevent cancer or not, in this study, blood was collected from residents in the Misasa hot spring district and in a control district. The level of a representative cancer-suppressive gene, p53, and the activity of a representative antioxidant enzyme, superoxide dismutase (SOD), were analyzed as indices. The level of serum p53 protein in the males in the Misasa hot spring district was found to be 2-fold higher than that in the control district, which is a significant difference. In the females in the Misasa hot spring district, SOD activity was approximately 15% higher than that in the control district, which is also statistically significant, and exceeded the reference range of SOD activity despite advanced age. These results suggested that routine exposure of the residents in the Misasa hot spring district to radon at a concentration about 3 times higher than the national mean induces trace active oxygen in vivo, potentiating products of cancer-suppressive gene and antioxidant function. As the p53 protein level was high in the residents in the Misasa hot spring district, apoptosis of cancer cells may readily occur. PMID- 15802856 TI - Analysis of micronuclei in the transferrin-receptor positive reticulocytes from peripheral blood of nasopharyngeal cancer patients undergoing radiotherapy by a single-laser flow cytometer. AB - The automated micronucleus test is now accepted as a simple, objective, and accurate method for evaluating potential mutagenic effects caused by physical, chemical or biotic factors. This paper describes a single-laser flow cytometry, based on an immunomagnetic isolation technique in combination with acridine orange staining, to detect frequencies of micronucleated transferrin-receptor positive reticulocytes from human peripheral blood. Using this flow cytometric system, we detected the frequencies of micronucleated transferrin-receptor positive reticulocytes from 10 nasopharyngeal cancer patients undergoing radiotherapy and the baseline of the frequencies of micronucleated transferrin receptor positive reticulocytes from 7 healthy donors. The results showed that the mean frequency of micronucleated transferrin-receptor positive reticulocytes from healthy donors was 0.236% and that from nasopharyngeal cancer patients before radiotherapy was 0.297%. After radiotherapy it was significantly elevated. When the cumulative dose of radiotherapy was about 20Gy, it reached a maximum of 6.905%, and then, as the cumulative dose of radiotherapy continued to increase to 30Gy, 40Gy and 50Gy, the frequency decreased to 6.258%, 5.119% and 5.007% respectively. Our results indicated that the single-laser flow cytometric system was quick, reasonable and acceptable for detecting the frequency of micronucleated transferrin-receptor positive reticulocytes from human peripheral blood. PMID- 15802857 TI - Effect of tocopherol-monoglucoside (TMG), a water-soluble glycosylated derivate of vitamin E, on hematopoietic recovery in irradiated mice. AB - A preparation of alpha-tocopherol monoglucoside (TMG) administered i.p. at a dose of 600 mg/kg immediately after whole body gamma irradiation was examined for its radioprotective efficacy towards bone marrow and peripheral blood nucleated cells. When mice received X-rays at a dose of 5,6 Gy, a marked decrease in bone marrow karyocytes and a reduction of peripheral leukocytes within the early post irradiated period were observed. However these changes were attenuated in TMG treated mice. Significant protection of blood lymphocytes was found for the TMG group of mice. The return to normal value of the reduced blood leukocyte count starting from the 8th day was more rapid in TMG-treated mice than in untreated irradiated mice. TMG administration was found to enhance hematopoietic recovery, as measured by the exceeded nucleated bone marrow cell count due to elevated amount of both lymphoid and granulocytic elements in the TMG-group, in comparison with that of both control irradiated and non-irradiated animals. These findings indicate that the radioprotective effect of TMG is apparently realized through its influence on hematopoietic system. PMID- 15802858 TI - Potential in a single cancer cell to produce heterogeneous morphology, radiosensitivity and gene expression. AB - Morphologically heterogeneous colonies were formed from a cultured cell line (KYSE70) established from one human esophageal carcinoma tissue. Two subclones were separated from a single clone (clone13) of KYSE70 cells. One subclone (clone13-3G) formed mainly mounding colonies and the other (clone 13-6G) formed flat, diffusive colonies. X-irradiation stimulated the cells to dedifferentiate from the mounding state to the flat, diffusive state. Clone 13-6G cells were more radiosensitive than the other 3 cell lines. Clustering analysis for gene expression level by oligonucleotide microarray demonstrated that in the radiosensitive clone13-6G cells, expression of genes involved in cell adhesion was upregulated, but genes involved in the response to DNA damage stimulus were downregulated. The data demonstrated that a single cancer cell had the potential to produce progeny heterogeneous in terms of morphology, radiation sensitivity and gene expression, and irradiation enhanced the dedifferentiation of cancer cells. PMID- 15802859 TI - Biological gain of carbon-ion radiotherapy for the early response of tumor growth delay and against early response of skin reaction in mice. AB - The biological effectiveness of carbon ions relative to gamma rays (RBE) was compared between the tumor growth delay and an early skin reaction of syngeneic mice. The RBE was larger for a tumor than skin when irradiated with large doses of high-LET (linear energy transfer) carbon ions. The intra-track damage (a term of a linear quadratic model) of a tumor and skin increased equally with an increase of the LET, while the inter-track damage (beta term) of skin alone increased with the LET. These data provide evidence that high-LET radiotherapy could achieve therapeutic gain by minimizing the difference in response to fractionated irradiation between the tumor and normal tissue. PMID- 15802860 TI - Influence of seed extract of Syzygium Cumini (Jamun) on mice exposed to different doses of gamma-radiation. AB - The radioprotective activity of the hydroalcoholic extract of jamun seeds (SCE) was studied in mice exposed to different doses of gamma radiation. The mice were injected with 0, 5, 10, 20, 40, 60, 80, 100, 120, 140 or 160 mg/kg body weight of SCE, before exposure to 10 Gy of gamma radiation, to select the optimum dose of radiation protection. The 80 mg/kg SCE was found to offer highest protection, therefore, further studies were carried out using this dose. The drug was more effective when administered through the intraperitoneal route at equimolar doses than the oral route. Since higher survival was observed for the i.p. route (50%), than the oral route (29.2%), all other studies were carried out by injecting SCE intraperitoneally. The mice treated with 80 mg/kg body weight SCE intraperitoneally before exposure to 6, 7, 8, 9, 10 and 11 Gy of gamma radiation showed reduction in the symptoms of radiation sickness and mortality at all exposure doses and caused a significant increase in the animal survival when compared with the concurrent double distilled water (DDW) + irradiation group. The SCE treatment protected mice against the gastrointestinal as well as bone marrow deaths and the DRF was found to be 1.24. PMID- 15802861 TI - Radio-sensitivity of the cells from amyotrophic lateral sclerosis model mice transfected with human mutant SOD1. AB - In order to clarify the possible involvement of oxidative damage induced by ionizing radiation in the onset and/or progression of familial amyotrophic lateral sclerosis (ALS), we studied radio-sensitivity in primary cells derived from ALS model mice expressing human mutant SOD1. The primary mouse cells expressed both mouse and the mutant human SOD1. The cell survival of the transgenic mice (with mutant SOD1), determined by counting cell numbers at a scheduled time after X-irradiation, is very similar to that of cells from wild type animals. The induction and repair of DNA damage in the transgenic cells, measured by single cell gel electrophoresis and pulsed field gel electrophoresis, are also similar to those of wild type cells. These results indicate that the human mutant SOD1 gene does not seem to contribute to the alteration of radio sensitivity, at least in the fibroblastic cells used here. Although it is necessary to consider the difference in cell types between fibroblastic and neuronal cells, the present results may suggest that ionizing radiation is not primarily responsible for the onset of familial ALS with the SOD1 mutation, and that the excess risks are probably not a concern for radiation diagnosis and therapy in familial ALS patients. PMID- 15802862 TI - Effects of high linear energy transfer radiation on the cochlea of C3H/He mouse during postnatal developmental course. AB - To investigate the biological effectiveness of neutrons at energies below 1MeV with regard to ear toxicity, we exposed mice to 1.0 Gy of monoenergetic neutrons (1.026 MeV) or 137Cs gamma rays at 7 days of age, and observed subsequent morphological changes in the inner ear with light and scanning electron microscopes. Monoenergetic neutrons, but not gamma rays, caused acute changes in the ear. The epithelium of the greater epithelial ridge in the organ of Corti showed degeneration around 6 hours and disappeared by 72 hours post-irradiation. The apoptotic cell death of the epithelium of the greater epithelial ridge was inducible by the radiation at 3 or 4 days of age. The hair cells formed the protrusion structures of the surface by 72 hours post-irradiation. Neutron irradiation also caused acute otitis media until 10 weeks of age. PMID- 15802863 TI - Intestinal tumorigenesis in Min mice is enhanced by X-irradiation in an age dependent manner. AB - We examined the effect of X-irradiation on intestinal tumorigenesis in Min (multiple intestinal neoplasia) mice. Single whole-body irradiation was given to mice of various ages from newborn to young adults. On the C57BL/6J (B6) background, X-irradiation increased tumor multiplicity of the small intestine exposed at ages from 2-3 days to 24-25 days, with a peak of 2.7-fold increase at 10-12 days of age; exposure at later ages resulted in only a slight increase. X irradiation also increased colonic tumors; however, the susceptible age period appeared earlier than that of the small intestine; the peak value of 4.6-fold increase was observed in the exposure at around 2-3 days of age. Irradiation at 24 days or later ages showed almost no effect on the colonic tumor induction. On the (B6 x MSM)F1 background, X-irradiation resulted in 2.7-fold increase in the small intestinal tumors, but no increase in the colonic tumors, and besides, the age dependency observed in the small intestinal tumors was much attenuated. Collectively, we conclude that tumorigenic efficacy of X-irradiation in Min mice was determined by the combination of the target organ, the age at exposure, and the genetic background. PMID- 15802864 TI - Real-time imaging of hypoxia-inducible factor-1 activity in tumor xenografts. AB - Hypoxia-inducible factor-1 (HIF-1) is responsible for various gene expressions related to tumor malignancy, such as metastasis, invasion and angiogenesis. Therefore, monitoring HIF-1 activity in solid tumors is becoming increasingly important in clinical and basic studies. To establish a convenient system for visualizing HIF-1 activity in tumor xenografts, we employed a promoter consisting of five copies of hypoxia response elements (5HRE), whose activity depends on HIF 1, and used a derivative of green fluorescence protein (d2EGFP) as a reporter gene. A human melanoma cell line, Be11, which contains the 5HRE-d2EGFP gene, showed fluorescence in response to hypoxia. The fluorescent intensity correlated inversely with the surrounding oxygen tension, and was time-dependent for the hypoxic treatment. Reoxygenation resulted in a rapid decrease in fluorescence due to the signal sequence for protein degradation encoded in d2EGFP, which enabled monitoring of HIF-1 activity in real-time. Heterogeneous fluorescence was observed in the solid tumor of a non-sacrificed tumor-bearing mouse. Immunohistochemical analysis confirmed that d2EGFP-expressing regions overlapped with the ones stained with a hypoxia marker, pimonidazole. These results suggest that the 5HRE-d2EGFP gene is suitable for the real-time imaging of HIF-1 activating cells in vivo, due to the short half-life of the d2EGFP protein as well as the specificity of the 5HRE promoter for HIF-1 activity. PMID- 15802865 TI - Enhanced induction of apoptosis by combined treatment of human carcinoma cells with X rays and death receptor agonists. AB - The death receptors Fas and DR5 are known to be expressed not only in immune cells but also in various tumor cells. The aim of the present study was to determine whether X irradiation enhanced induction of apoptosis in Tp53 wild type and Tp53-mutated tumor cell lines treated with agonists against these death receptors. We showed that 5 Gy of X irradiation significantly up-regulated the expression of death receptors Fas and DR5 on the plasma membrane in gastric cancer cell lines MKN45 and MKN28, lung cancer cell line A549, and prostate cancer cell line DU145, and that subsequent treatments with agonistic molecules for these death receptors, Fas antibody CH11 and TRAIL, increased the formation of active fragment p20 of caspase 3 followed by the induction of apoptosis. This death-receptor-mediated apoptosis was independent of Tp53 status since MKN28 and DU145 were Tp53-mutated. The post-irradiation treatment of the cells with N acetyl-L-cysteine (NAC) abolished the up-regulation of the expression of Fas and DR5 on the plasma membrane. NAC also attenuated the increase in the formation of p20 and the induction of apoptosis by agonistic molecules. These results suggested that the increase in the induction of apoptosis by combined treatment with X irradiation and CH11 or TRAIL occurred through a change of the intracellular redox state independent of Tp53 status in human carcinoma cell lines. PMID- 15802866 TI - Cytogenetic radiosensitivity of g0-lymphocytes of breast and esophageal cancer patients as determined by micronucleus assay. AB - Enhanced chromosomal radiosensitivity is a feature of many cancer predisposition conditions, indicative of the important role of chromosomal alterations in carcinogenesis. In this study the cytokinesis-blocked micronucleous assay was used to compare the radiosensitivity of blood lymphocytes obtained from Iranian breast or esophageal cancer patients (n = 50, n = 16; respectively) with that of control individuals (n = 40). For each sample, one thousand binucleate lymphocytes were analyzed before and after in vitro exposure to 3 Gy of gamma rays. The radiation-induced frequency of micronucleus was significantly higher in the breast cancer group (261/1,000 binucleated cells) than in esophageal cancer group (241/1,000 binucleated cells, P < 0.01) or in the control group (240/1,000 binucleated cells, P < 0.01). The results indicate that breast cancer patients are more radiosensitive compared to normal healthy individuals or esophageal cancer patients. Increased radiosensitivity could be due to defects in DNA repair genes involved in breast cancer formation. Since patients with esophageal cancer did not show elevated radiosensitivity, it is assumed that the contribution of radiosensitivity-related genes to the development of esophageal cancer may be smaller than the contribution of those genes to breast cancer. PMID- 15802867 TI - Glycine betaine, a beer component, protects radiation-induced injury. AB - Human whole-blood was exposed to 137Cs gamma-rays or 50 keV/microm carbon ions in the presence or absence of glycine betaine, a beer component in vitro. The dicentrics of chromosome aberrations in human lymphocytes were significantly (p < 0.05) reduced by glycine betaine after irradiation with 4 Gy of either gamma-rays or carbon ions. The maximum protection by glycine betaine for gamma-rays or carbon ions was 37% and 20%, respectively. C3H/He female mice, aged 14 weeks, received an i.p. injection of glycine betaine 15 min before whole-body irradiation with gamma-rays or 50 keV/microm carbon ions. Glycine betaine significantly (p < 0.05) increased the percent survival of irradiated mice with either gamma-rays or carbon ions. In conclusion, glycine betaine is a potent protector against damages caused by low- and high-LET radiation. PMID- 15802868 TI - Mortality of british radiologists: a lecture note. AB - The precautions introduced after the first 23 years experience of the use of x rays for medical diagnosis proved adequate to eliminate the acute hazards of exposure, but it was much longer before it was realized that small doses that did not produce any acute effect could increase the risk of cancer. British radiologists who took up the specialty at different periods have, therefore, been studied to see if the risk has now been adequately controlled. Four groups have been studied starting respectively before 1921, in 1921-34, 1935-54, and 1955-77, corresponding approximately to periods when different limits of exposure were applied. Altogether 2698 male radiologists have been identified and all but 27 followed successfully to emigration, death, or survival to January 1st 1997. Of the 1198 who had died, 228 are known to have died of cancer. Two problems arise in evaluating the carcinogenic hazard to which they were exposed: the assessment of the doses received and the selection of an appropriate control group with which to compare their mortality. The most appropriate comparison group would seem to be medical practitioners in general. In comparison with them, radiologists entering in the first 3 periods had increased risks of death from cancer though appreciably less than would have been predicted from the expected effect of the radiation they had received. Those who joined in the latest period had a relatively reduced risk, irrespective of any effect of the small dose of radiation they are likely to have received. Independent evidence suggests, however, that since 1951 radiologists have smoked less than other doctors and the lower than predicted risk in the groups exposed since 1920 is limited to smoking related cancers, the mortality from other cancers being higher than in doctors generally. In assessing the risk of occupational exposure to radiation, life style has to be taken into consideration, as well as dose of radiation. PMID- 15802869 TI - Urinary tract infection aggravates oxidative stress in diabetic patients. AB - To investigate the effect of urinary tract infection on oxidative stress in diabetic patients, we measured the activities of antioxidant enzymes such as catalase and superoxide dismutase, and lipid peroxidation levels in urine specimens of type II diabetic patents with urinary tract infection. A total of 69 patients were included into this study: 23 non-diabetic patients with urinary tract infection, 28 patients with diabetes mellitus, and 18 diabetic patients with urinary tract infection. Twenty-five healthy subjects, matched for age, sex, body mass index and smoking status were also included as control. Urine cultures were performed by the standard techniques, and all grown bacteria were identified as Escherichia coli. Antioxidant enzymes and lipid peroxidation levels in urine were measured by spectrophotometric method. In urine samples of diabetic patients with or without urinary tract infection and in urine samples of non-diabetic patients with urinary tract infection, catalase and superoxide dismutase activities were lower and lipid peroxidation levels were higher than those of the healthy subjects (p < 0.05). Diabetic patients without urinary tract infection were similar to non-diabetic patients with urinary tract infection. Decreased antioxidant capacity and the increased levels of lipid peroxidation were profoundly higher in diabetic patients with urinary tract infection. These results indicate that urinary tract infection aggravates the oxidative stress in diabetic patients. Therefore we believe that diabetic patients with urinary tract infection need antioxidant treatment. PMID- 15802870 TI - The effect of tirofiban and clopidogrel pretreatment on outcome of old saphenous vein graft stenting in patients with acute coronary syndromes. AB - In spite of developments in interventional cardiology, the success rate of saphenous vein graft stenting is still low in patients with acute coronary syndromes. In this study, we aimed at finding out the effect of pretreatment with Tirofiban, a glycoprotein IIb/IIIa inhibitor, and clopidogrel, an adenosine diphosphate antagonist, on the outcome of saphenous vein graft stenting in patients with acute coronary syndrome. A total of 47 patients, who had lesions in saphenous vein grafts and acute coronary syndrome, could be randomized to treated group (n = 24), who received Tirofiban and clopidogrel for 48 hours before the intervention, and untreated group (n = 23), who did not receive Tirofiban and clopidogrel. In the untreated group, the intervention was performed just after the coronary angiography. All patients underwent stenting as the standard intervention. The groups were compared by Mann-Whitney's U-test or Chi-Square test. The level of statistical significance was set at 0.05. There were no significant differences regarding age, gender, and atherosclerotic risk factors between the two groups. In treated group, precutaneous coronary intervention was successful in all patients and no-reflow phenomenon occurred in only one patient. The rate of no-reflow or slow-flow phenomenon was significantly lower in treated group (one patient vs 9 patients, p = 0.004). One patient in untreated group experienced ventricular fibrillation, which was converted to sinus rhythm after defibrillation. During short-term follow-up, there were no acute myocardial infarction, coronary bypass surgery or death in both groups. There was no major bleeding. Minor bleeding was more frequent in treated group, but it did not achieve statistical significance (3 vs 1; p = 0.322). In conclusion, pretreatment with tirofiban and clopidogrel before percutaneous coronary intervention might result in better immediate outcomes in old saphenous vein grafts without any significant increase in bleeding complications. PMID- 15802871 TI - Cognitive event-related potential and neuropsychological findings in Behcet's disease without neurological manifestations. AB - Behcet's disease (BD) is a multisystem inflammatory disorder characterized by recurrent oral and genital ulcers and uveitis. BD patients without neurological involvement frequently have mild neurological symptoms. The aim of this study was to evaluate whether BD patients without neurological involvement have any changes in cognitive functions. Twenty BD patients without neurological involvement and 13 control subjects were included in the study and were analyzed by neurophysiological and neuropsychological examinations. The cognitive event related potentials (P300) were recorded from the frontal, central and parietal areas of the right and left hemispheres of the patients and control subjects. Likewise, all individuals were evaluated with neuropsychological tests. In contrast to a study with similar design, we did not find any difference between the cognitive event-related potentials values of BD patients without neurological involvement and the control subjects. All BD patients without neurological involvement exhibit normal results of the neuropsychological test. In conclusion, the results of neuropsychological tests and cognitive event-related potentials values in BD patients without neurological involvement are indistinguishable from those in control subjects and no alteration of cognitive functions is present. PMID- 15802872 TI - Liver fat content measured by magnetic resonance spectroscopy at 3.0 tesla independently correlates with plasminogen activator inhibitor-1 and body mass index in type 2 diabetic subjects. AB - We measured liver fat content by 3-Tesla magnetic resonance spectroscopy (MRS) in 34 non- to mild obese Japanese subjects with type 2 diabetes, who were not complicated with any liver diseases including clinical fatty liver (liver/spleen ratio of computed tomography [CT] < 0.9) and were not being treated with oral hypoglycemic agents, insulin, or lipid-lowering agents, and analyzed the relationship between liver fat content and body composition and plasma metabolite. The liver fat content is significantly correlated with variables relating to obesity (body mass index [BMI], body weight, fat mass, waist to hip ratio, visceral fat area, subcutaneous fat area, and serum triglyceride), insulin resistance (fasting plasma insulin and homeostasis model assessment of insulin resistance [HOMA-IR]), adipocytokines (serum plasminogen activator inhibitor-1 [PAI-1] and leptin), and serum cholinesterase, but not CT liver/spleen ratio, which is correlated only with fasting plasma glucose, BMI, and HOMA-IR. Multiple regression analysis revealed that the liver fat content is independently associated with serum PAI-1 level (p < 0.001) and BMI (p < 0.05), but not visceral fat area. MRS is a more sensitive method for quantifying liver fat content than CT in type 2 diabetic subjects with non- to mild obesity and without clinical fatty liver. PMID- 15802873 TI - Ber-EP4 and anti-calretinin antibodies: a useful combination for differential diagnosis of various histological types of ovarian cancer cells and mesothelial cells. AB - The differential diagnosis between reactive mesothelial cells and ovarian carcinoma cells is often difficult in cytologic specimens. Immunocytochemical procedures have been utilized in assisting this differential diagnosis, with limitations. Furthermore, previous studies examined only serous type but not other histological types of ovarian carcinoma cases. Therefore, we evaluated the practical value of various epithelial and mesothelial markers in differential diagnosis of these two types of cells. Various types of ovarian carcinoma (serous, n = 22; mucinous, n = 10; endometrioid, n = 7; clear cell, n = 10) and benign mesothelial tissues (n = 15) were studied by immunohistochemistry. We then studied effective panels of antibodies by immunohistochemistry in 43 cytologic specimens of ascites or peritoneal lavage fluid consisting of 20 reactive mesothelium and 23 adenocarcinomas of the ovary. In the tissue specimens, Ber EP4, a monoclonal antibody of epithelial antigen, and a polyclonal antibody against calretinin, which is expressed in mesothelium, are used in differentiating reactive mesothelial cells from ovarian carcinoma. In cytologic specimens, the sensitivity and specificity of Ber-EP4 were 100% and 90%, respectively. The sensitivity and specificity of the anti-calretinin antibody were 90% and 91%, respectively. Using multiple regression analysis, the correlation coefficient between epithelial antigen and calretinin reactivity was r = 0.938, with a significance level of p < 0.0001. In conclusion, the combined immunostaining of cytologic specimens for Ber-EP4 and the anti-calretinin antibody is helpful for the differential diagnosis between mesothelial cells and not only serous type, but also mucinous, endometrioid and clear cell types of ovarian cancer cells. PMID- 15802874 TI - Validity and utility of the Craig Hospital Inventory of Environmental Factors for Korean community-dwelling elderly with or without stroke. AB - The social environmental barriers are considered to be important because the "social participation" of people with impairments would be facilitated by the prevention and reduction of environmental barriers. The Craig Hospital Inventory of Environmental Factors (CHIEF) is one of the few scales to assess the environmental barriers. In this study, we developed the Korean version of CHIEF and evaluated its construct validity and utility in a sample of Korean community dwelling elderly with or without stroke. We evaluated the construct validity of the CHIEF by testing the original five-factor structure using a confirmatory factor analysis in 400 elderly in Seoul, Korea. The utility of the CHIEF was then assessed by examining the relationships between individual characteristics, Barthel Index and perceived environmental barriers, measured by the CHIEF, using a structural equation modeling approach. The confirmatory factor analysis result demonstrated the validity of a second-order factor model of the CHIEF comprising the five factors as first-order factors. The perceived environmental barrier was a second-order factor when provided acceptable fit indices after two modifications. The structural equation modeling indicates that perceived environmental barriers are significantly related to activities of daily life but not age, gender, and the episode of stroke. The CHIEF is useful in measuring environmental factors for Korean older adults with or without stroke. PMID- 15802875 TI - Prevalence and characteristics of migraine in women of reproductive age in Istanbul, Turkey: a population based survey. AB - Migraine is more common in female and onset of migraine is most commonly seen in the second and third decades of life. In this study, we aimed to estimate the prevalence and characteristics of migraine among women of ages between 15 to 45 years in Turkey. This is the first study to target this age group. The women were interviewed on a door-to-door basis, from early morning to late evening. Once responded positively to headache, an in-depth interview was performed questioning for migraine features. Diagnosis was made from a questionnaire by eight neurologists. One thousand eight hundred thirty five (1,835) out of 96,000 women living in Maltepe which is a town of Istanbul participated in this study. The prevalence of migraine in females aged 15-45 (reproductive ages) was 15.8% (95% CI, 0.142-0.176). This study showed that migraine onset occurred at a mean age of 22.7, 33% having family history, and with migraine with and without aura having near equal frequencies. The prevalence of migraine in women of reproductive ages in Istanbul as found in our study is lower than that reported in United States and Europe countries, but higher than that in Middle and Far Eastern countries. PMID- 15802876 TI - Differential growth properties of normal and malignant esophageal epithelial cells: a possible cross talk between transforming growth factor-beta1 and epidermal growth factor signaling. AB - Comparative cultures of normal and malignant cells are important for understanding the growth properties of tumors. Although many cell lines have been established from esophageal cancers, the growth properties of normal and cancer derived esophageal epithelial cells have not been compared extensively. We succeeded in establishing an assay system in serum-free conditions for normal human esophageal epithelial cells (HEE cells) and 14 cancer-derived esophageal epithelial cell lines (TE-cell lines). The growth properties of these cells were characterized upon stimulation with various growth-related factors. Among these factors, acidic fibroblast growth factor (aFGF) was the most effective stimulant for both the HEE cells and all the TE-cell lines. Most TE-cell lines required a higher concentration of calcium for their growth than did the HEE cells. Transforming growth factor-beta1 (TGF-beta1) inhibited the growth of HEE cells and 7 TE-cell lines; however, the other 7 TE-cell lines were resistant to the inhibitory effect of TGF-beta1. Interestingly, epidermal growth factor (EGF) had a much greater stimulatory effect on the TGF-beta1-resistant cells than the TGF beta1-sensitive cells. Although ethanolamine enhanced the growth-promoting ability of EGF or aFGF in the TGF-beta1-sensitive cells, it had no effect on the TGF-beta1-resistant cells. These findings suggested a possible cross talk between TGF-beta1 and EGF signaling, and an important role of ethanolamine in the signaling pathways of growth factors. This serum-free culture system will contribute to clarify the altered signaling pathways of esophageal cancer. PMID- 15802877 TI - Medicolegal childhood deaths in Adana, Turkey. AB - The objective of this study is to determine the characteristics of and changes in the medicolegal childhood deaths that occurred in the provincial center of Adana, and to contribute to the establishment of a database for the development of national and international policies. The study is a retrospective research examining the 1,110 cases in the age group 0 to 18, which were specified as medicolegal deaths in the provincial center of Adana-Turkey between the years 2000-2004. The cases were examined according to age, sex, causes of deaths, the origins and scenes of occurrence. Among all the deaths, 523 cases (47.2%) were seen in the age group of 0-6 years. Accidents account for 900 cases (81.1%), and blunt traumas (594 cases) represent the most common cause of deaths. The share of traffic accidents in total blunt traumas was found to be 441 cases (74.2%). And 270 (24.3%) of all medicolegal deaths occurred as a result of household accidents. The present study has also indicated that the home and surroundings represent the highest-risk areas. As a conclusion, setting up childhood injury prevention committees and providing training programs for parents will be effective for the prevention of childhood injuries and deaths. PMID- 15802878 TI - Vasculo-Behcet's disease mimicking a metastatic neck mass. AB - This article presents a case of Behcet's Disease (BD) with vascular involvement of the neck, which mimicks a metastatic neck mass in the initial presentation. A 58-year-old man presented with dysphagia, weight loss, bulging on the lateral wall of the left pyriform sinus, and a firm and fixated neck mass suggestive of metastasis. Computed tomography of the neck demonstrated a solid mass, around the bifurcation of the carotid artery together with a pseudoaneurysm of the left external carotid artery. The mass was about four centimeters in diameter and extended to hypopharynx medially. Biopsy from neck mass and hypopharynx revealed no specific pathology. During follow-up the firm and fixated mass changed into a completely pulsatile one in the following three weeks. Reassessment of the patient's past history in detail revealed that he had had recurrent oro-genital ulcers, arthralgia and recurrent skin lesions. The pathergy test was positive. The patient was diagnosed to be BD and treatment consisting of colchicine 1 mg/day, peroral was started. He had a favorable outcome after treatment and was asymptomatic at follow-up of 24 months. It is unusual for BD to present as a neck mass but yet it must be considered in the differential diagnosis of neck masses. The present case report demonstrates how such a mass may mimic metastatic tumoral involvement and cause diagnostic dilemma. PMID- 15802879 TI - From physical pharmacy to clinical pharmacology. AB - Research works on molecular interactions in solutions were carried out at School of Pharmacy, the University of Wisconsin under the direction of Prof. T. Higuchi and at Faculty of Pharmaceutical Sciences, Kyoto University under the direction of Prof. H. Sezaki. Studies on permeation of drugs through polymer membranes were carried out at Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Canada and at Pharmaceutical Chemistry Research Laboratories at Food and Drug Directorate, Department of Health and Welfare, Canada. Studies on modification of delivery patterns by means of pharmaceutical approaches were carried out at Faculty of Pharmaceutical Sciences, Hokkaido University. Topics related to modification of drug delivery patterns include employment of amorphous forms such as ground mixture with micro-crystalline cellulose and coprecipitate with polyvinylpyrrolidone, use of biodegradable polymers such as polylactic acid and polycarbonates, gel-forming materials such as konjac, agar and hydroxypropylcellulose, and physicochemical systems such as complexation. Works related to drug delivery and disposition of drugs in humans were carried out at Department of Pharmacy, Kumamoto University Hospital. Topics related to drug delivery in humans include injections containing anticancer drugs for intra arterial administration, lidocaine gels for dermal anesthesia, glucagon solution for nasal administration. Topics related to disposition of drugs in humans include clinical pharmacokinetic studies in infants and elderly and medical uses of adsorbents. PMID- 15802880 TI - [Pharmacological properties of Gyokuheifusan, a traditional Kampo medicinal formula]. AB - Gyokuheifusan (GHS, Jade Windscreen Powder in English, Yupingfengsan in Chinese) is an herbal formula in traditional Kampo medicine which consolidates superficial resistance to protect against invasion by external pathogens. This review describes the immunopharmacologic properties of GHS as a holistic Kampo medicine, which can affect human homeostasis and constitution of human beings. Oral treatment with GHS has preventive and curative effects in allergic rhinitis induced by Japanese cedar pollen in guinea pigs. Since these effects do not occur with authentic antiallergic agents, GHS appears to have holistic effects on allergic rhinitis. In another study, the effects of GHS on murine antibody production against ovalbumin (OVA) were evaluated. When mice were sensitized intraperitoneally to OVA, the concentration of OVA-specific immunoglobulins in the sera significantly increased with GHS treatment. When they were sensitized intranasally to OVA, GHS significantly reduced the concentration of OVA-specific antibodies in the sera. It was suggested that GHS stimulates immune responses when the antigen had already invaded the body, and that GHS might consolidate the resistance of nasal mucosa to protect from OVA invasion, and then OVA-specific antibodies in sera might be suppressed. These results suggest that traditional medicines have own characteristics different from those of modern medicines, and that original pharmacologic experiments are important to evaluate traditional medicines scientifically. PMID- 15802881 TI - [Development of the method for analysis of the cytokine balance shift associated with diseases]. AB - A dual-color enzyme-linked immunospot (ELISPOT) assay enabled us to analyze three types of cytokine-secreting cells simultaneously. T helper (Th) cells can be subdivided into at least two distinct functional subsets based on their cytokine secretion profiles. The first type of clones (Th1) produces interleukin-2 (IL-2) and interferon-gamma (IFN-gamma), but not IL-4 or IL-5. The second type of clones (Th2) produces IL-4 and IL-5, but not IL-2 or IFN-gamma. Furthermore, the presence of the third type (Th0) of cell, which is a precursor of Th1 or Th2 cells, has been demonstrated to produce both Th1- and Th2-type cytokines. The dual-color ELISPOT assay was developed to differentiate these three subtypes of Th cells in an identical well. In the system, the red spots corresponding to IL-2 secreting cells (Th1) were developed with horseradish peroxidase and aminoethylcarbazole/H(2)O(2). The light-blue spots corresponding to IL-4 secreting cells (Th2) were developed with alkaline phosphatase and Vector blue (chromogenic substrate for alkaline phosphatase). The mixed-colored (indigo) spots corresponding to both types of cytokine-secreting cells (Th0-cells) were developed with both chromogenic substrates. With this system, we could detect the IL-2- and/or IL-4-secreting cells simultaneously in a murine spleen cell or human peripheral mononuclear cell preparation. PMID- 15802882 TI - [Effects of Chinese herbal medicines on intestinal drug absorption]. AB - Sho-saiko-to (Xiao-Chai-Hu-Tang), one of the major traditional Chinese medicines, has been frequently prescribed with other synthetic or biotechnological drugs for the treatment of various acute or chronic diseases in Japan, and thus it is important to understand the interactions between Sho-saiko-to and coadministered drugs. This paper reviews the effects of Sho-saiko-to on the pharmacokinetics and pharmacodynamics of concomitant drugs in the gastrointestinal tract. Sho-saiko-to slightly hastens the gastrointestinal absorption of the sulfonylurea compound tolbutamide. Furthermore, it is considered that the increase in the gastrointestinal absorption rate by Sho-saiko-to may potentiate the hypoglycemic effects of tolbutamide in the early period after oral administration. Sho-saiko to can facilitate the epithelial membrane permeability of tolbutamide at an early phase across the rat jejunum in situ and Caco-2 cell monolayers. It is also suggested that Sho-saiko-to enhances the energy-dependent transport of tolbutamide and has an inhibitory effect on the passive paracellular transport of tolbutamide in Caco-2 cells. This result might be related to the accelerated in vivo absorption rate of tolbutamide by concomitant dosing with Sho-saiko-to in rats. In addition, Sho-saiko-to has inhibitory effects on the efflux pump mediated by MDR1, and it appears that the crude constituents in Glycyrrhizae radix, glycyrrhizic acid and liquiritin, contribute to MDR1 suppression. PMID- 15802883 TI - DPPH radical-scavenging effect of several phenylpropanoid compounds and their glycoside derivatives. AB - Eugenol, isoeugenol, caffeic acid, ferulic acid, isoferulic acid, estragole, trans-anethole, and paeonol are components of a Chinese herbal medicine used as a painkiller and stomachic. We investigated the potential role of these compounds as antioxidants. We studied the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging effect of these molecules, together with some glycoside derivatives, to ascertain their potential in reducing the levels of activated oxygen species in vivo. The DPPH radical-scavenging effects of eugenol, isoeugenol, and the glycoside derivatives of caffeic acid, ferulic acid, and isoferulic acid (SC(50)=8-28 microM) were similar to those of alpha-tocopherol, which was used as a positive control. PMID- 15802884 TI - Statistical approach to drug-induced taste disorders based on zinc chelating ability. AB - Various drug-induced taste disorders have been ascribed to zinc deficiency in serum. Assuming that the zinc deficiency is caused by the chelating reaction of zinc ions with drugs, the electrode potential of the Zn(2+)/Zn(Hg) system was measured in the presence of drugs in water, ethanol, and N,N-dimethylformamide (DMF). The zinc-chelating ability was estimated based on the potential change Delta E(2) with the addition of a drug. A large potential change suggesting potent chelating ability was observed in penicillamine, furosemide, and ibuprofen in ethanol and in fluorouracil, acetazolamide, and bezafibrate in DMF. Multiple regression analysis was used to evaluate the observed Delta E(2) in mV to represent chelating ability. The regression equation to estimate the frequency of taste disorders was deduced from Delta E(2), and frequency of four drugs appeared in package inserts and interview forms. According to the regression equation, the frequency of taste disorders was successfully estimated for 14 drugs examined in this study. The result was examined in a clinical case. PMID- 15802885 TI - [Radical formation by grinding of commercial tablets according to hospital and pharmacy prescription]. AB - We examined mechanoradical formation in the grinding process of commercial tablets using electron spin resonance (ESR). Mechanoradicals were detected in all tested samples (23 types of commercial tablets) when the ball-milling of tablets was conducted under anaerobic conditions and some were fairly stable even in air. Thus the grinding may cause changes in the physicochemical properties of ingredients included in commercial tablets. Because high quality is demanded in pharmaceuticals, these results suggest more caution should be taken in the grinding of commercial tablets in hospitals and pharmacies. PMID- 15802886 TI - [Gemcitabine in the first line therapy of advanced and metastatic non-small-cell lung carcinoma (NSCLC): review of the results of phase III studies]. AB - Modern platinum-based combination therapies containing gemcitabine, vinorelbine or taxanes produce response rates of 30-40%, median survival times of 8-10 months and 1-year survival rates of approximately 35% in patients with advanced non small-cell lung cancer (NSCLC). Of the new drugs available, gemcitabine (Gemzar, Lilly, Bad Homburg, Germany) has been the most extensively researched in clinical trials and exhibits a consistent database. A total of 37 randomized phase III trials involving more than 15,000 patients have been published to evaluate gemcitabine as first-line therapy for treating locally advanced and/or metastatic NSCLC. One trial studied gemcitabine exclusively as a single agent and another four trials investigated the drug in monotherapy and combination therapy. Of the 36 combination treatment studies, 21 included gemcitabine plus cisplatin treatment arms, 6 investigated gemcitabine plus carboplatin, and another 12 evaluated platinum-free gemcitabine combinations with other third generation cytostatic agents (multiple nominations possible). In single-agent treatment, gemcitabine was similarly effective to older platinum-based combinations such as vindesine-cisplatin but was less toxic. Thrombocytopenia was the main dose limiting toxicity but was rarely clinically relevant. A 3-week cycle with gemcitabine on days 1 and 8 was confirmed as being the most convenient of the gemcitabine-based combinations studied. No other modern platinum-based doublet with vinorelbine or taxanes was superior to gemcitabine plus cisplatin in terms of survival or time to progression in any of the eight phase III studies performed. These results are consistent with previous phase II data and with a recent meta-analysis of 11 phase III and 2 randomized phase II studies involving more than 4,500 patients (1,861 in gemcitabine-based treatment arms). This meta analysis also demonstrated a statistically significant benefit regarding overall and progression-free survival for gemcitabine-platinum- based regimens compared with other platinum combinations. In summary, currently available data indicate that gemcitabine-platinum 2-agent combinations given in 3-week cycles may at present have the best benefit-risk ratio in the treatment of advanced NSCLC. In contrast, platinum based 3-agent schedules do not offer any survival benefit. In elderly patients with poor performance status single agent treatment with a modern cytotoxic agent should be considered. Furthermore, according to the experiences from phase III studies so far, platinum- free combinations open up the possibility of a more feasible and clinically practical, active and well tolerated treatment which is associated with a positive impact on patient quality of life. PMID- 15802887 TI - Non-Hodgkin's lymphomas in Greece according to the WHO classification of lymphoid neoplasms. A retrospective analysis of 810 cases. AB - The purpose of this retrospective study, the largest unselected series in our country, was to illustrate the clinicopathological features of non-Hodgkin's lymphoma (NHL) classified according to the World Health Organization (WHO) classification of lymphoid neoplasms. A retrospective analysis was conducted and clinical features of histological subtypes were established in 810 patients (age > or = 15 years) with NHL who were treated at 8 major centers representative of Greece. There were 435 males and 375 females 95% of them aged >30 years. B symptoms were present in 34% of the patients, while 45.3% had stages I-II and 54.6% had stages III-IV. LDH was increased in 37% of the patients. B cell lymphomas formed 88% of the cases whereas T cell lymphomas formed 12% of the total. Indolent lymphomas accounted for 31.1%, aggressive ones for 66.7% and very aggressive ones for 2.4% of all NHLs. Among indolent lymphomas extranodal ones (MALT B cell lymphoma) were the most common subset while follicular lymphoma grade I and II and small lymphocytic ones presented with equal frequency. Among the aggressive lymphomas diffuse large cell lymphoma (DLCL) was the most common subtype; this entity along with large-cell immunoblastic lymphomas accounted for 45.2% of all B cell lymphomas. Among the T cell lymphomas, peripheral T cell lymphomas and anaplastic large cell lymphomas of the T/null-cell type were the most common subtypes. The most common extranodal presentation was the gastrointestinal tract (GI). Next in frequency were primary extranodal NHL of the head and neck region. MALT B cell lymphomas were found in almost half of the patients with GI tract NHL, whereas in all other extranodal places DLCL was the predominant histological subtype. The median survival for indolent and aggressive NHL was 123.5 and 55.5 months, respectively. This is the first report of a large series of malignant lymphomas in Greece using the WHO classification. It appears that there are no significant differences between NHL in Greece and other large series as far as clinical and extranodal presentation is concerned. The frequency of follicular lymphoma in the current study is comparable to that reported from Asian countries and mainland Europe, but lower than that of US and Northern European series. There were no important differences in the incidence of the remaining histological subtypes between Greece and other European countries. PMID- 15802888 TI - Haemopoietic cell transplantation of patients with a history of deep or invasive fungal infection during prophylaxis with liposomal amphotericin B. AB - Relapse of a preceding fungal infection is a considerable risk during haemopoietic stem cell transplantation. The optimal secondary prophylaxis has not been found so far since the application of standard drugs is hampered by potential ineffectiveness or intolerable side effects. This investigation describes haemopoietic cell transplantation of patients with a history of invasive or systemic fungal infection (IFI). The strategy was either administration of liposomal amphotericin B as secondary prophylaxis or an early switch to liposomal amphotericin B after administration of azoles. The 43 patients had a history of proven (n = 14), probable (n = 14) and possible (n = 15) IFI. Twenty-eight patients (65%) could be discharged from the BMT ward without signs of mycosis. Transplant-related mortality was 35%. Overall, 12 fungus-related (IFI) deaths (28%) occurred. The percentage of fungus-related deaths was highest in the 'proven' group with 43% compared to 20 and 21% in the two other groups. Side effects of liposomal amphotericin B were low. A discontinuation of the drug was not necessary in any patient. Serum creatinine showed a slight increase to 128% (median) of the baseline allowing continuous administration of concomitant nephrotoxic drugs such as cyclosporin A. In conclusion, secondary prophylaxis with or early switch to liposomal amphotericin B facilitates allogeneic stem cell transplantation of patients with a history of IFI with minor side effects. However, fungal infections and transplant-related mortality remain major problems in this often heavily pretreated subgroup of patients. PMID- 15802889 TI - Antiplatelet antibodies and their correlation with clinical findings in childhood immune thrombocytopenic purpura. AB - The demonstration of antiplatelet antibodies (PAIgG, PAIgM) and decreased detection of platelet surface antigens (CD41, CD61, CD42b) in children with immune thrombocytopenic purpura (ITP) have a diagnostic role. This study was conducted to determine whether these parameters differed in acute and chronic ITP. Chronic ITP was defined as thrombocytopenia persisting for more than 6 months from the onset of illness. A total of 80 subjects were divided into three groups: group 1 included 39 patients with acute ITP; group 2 included 31 patients with chronic ITP, and group 3 included 10 healthy children. At diagnosis, blood samples were obtained for platelet count, mean platelet volume, plateletcrit and platelet distribution width along with platelet surface antigens and antiplatelet immunoglobulins. We found that platelet surface antigens were significantly decreased in both acute and chronic ITP when compared to the control group (p = 0.001). In contrast, PAIgG was increased in acute and chronic ITP patients compared to the control group. PAIgM was significantly higher in acute ITP. We conclude that decreased platelet surface antigens and increased antiplatelet antibodies are observed in both acute and chronic ITP. In patients with chronic progress, a relatively lower level of PAIgM can be identified. PMID- 15802890 TI - Absence of gene mutation in TRAIL receptor 1 (TRAIL-R1) and TRAIL receptor 2 (TRAIL-R2) in chronic myelogenous leukemia and myelodysplastic syndrome, and analysis of mRNA Expressions of TRAIL and TRAIL-related genes in chronic myelogenous leukemia. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is an interferon (IFN)-induced molecule with apoptotic activity. We examined gene mutations in the death domains of TRAIL receptor 1 (TRAIL-R1) and TRAIL receptor 2 (TRAIL-R2), and in the TRAIL gene promoter in 46 chronic myelogenous leukemia (CML) patients. In 23 of the 46 patients, all the coding regions of TRAIL-R2 were also examined. However, no mutation or loss of heterozygosity was found. Furthermore, no mutation in the death domains of TRAIL-R1 and TRAIL-R2 genes, which causes amino acid change, was found in 18 myelodysplastic syndrome (MDS) patients. Ribonuclease protection assay (RPA) and real-time quantitative polymerase chain reaction using polymorphonuclear neutrophils of five new CML patients showed that the TRAIL mRNA expression was very low before in vitro IFN-alpha stimulation and markedly upregulated after IFN-alpha stimulation. FAS mRNA was also upregulated with IFN-alpha stimulation but the fold induction was far lower than that of TRAIL mRNA. In addition, RPA revealed that the ratio of (TRAIL-R1 plus TRAIL-R2) to TRAIL-R3 was also increased after IFN-alpha stimulation. Taken together, gene mutations of TRAIL-R1, TRAIL-R2 are infrequent in patients with CML and MDS. And so is the TRAIL promoter for CML. These mutations seem unrelated to tumorigenesis, disease progression, and response to IFN-alpha therapy in CML. A markedly high induction of TRAIL mRNA by IFN-alpha may have some relevance to IFN alpha action in CML patients. PMID- 15802891 TI - Macrophage activation syndrome in children with systemic-onset juvenile chronic arthritis. AB - Macrophage activation syndrome (MAS) is a life-threatening complication in children with rheumatic diseases, particularly systemic-onset juvenile chronic arthritis (SOJCA). Because of the potential fatality of this condition, prompt recognition and immediate therapeutic intervention are important. This study assessed the clinical features of nine MAS events in five children with SOJCA. Nonremitting fever and decreased platelet and white blood cell counts led to a diagnosis of MAS. The urinary beta2-microglobulin (beta2MG) level was a sensitive indicator of MAS. Serum levels of beta2MG and soluble interleukin-2 receptor were also elevated. These biologic markers reflecting hyperactivated cellular immunity are useful indicators of MAS. Four children treated with cyclosporin A (CSP) achieved rapid and complete recovery, but one patient without CSP died due to rapidly progressive respiratory failure. All children treated with CSP responded quickly, and fever abated within 36 h of initiation of treatment. CSP should be added to first-line therapy of MAS. PMID- 15802892 TI - Effect of cytokines and chemokines on sickle neutrophil adhesion to fibronectin. AB - A role for leukocytes in sickle cell vaso-occlusive crisis is becoming increasingly recognized. Neutrophil counts are higher in sickle cell patients and neutrophils from these patients demonstrate increased adhesion to endothelial monolayers under certain circumstances. The effects of selected cytokines on the adhesion mechanisms of normal neutrophils and neutrophils from sickle cell anaemia patients (SCA neutrophils) were investigated. Neutrophils were separated from the blood of homozygous (HbSS) SCA patients and healthy controls. Following pre-incubation (25 min, 37 degrees C) of the cells with cytokines, the adhesion of the cells to fibronectin (FN)-coated plates (20 micro) was determined (60 min, 37 degrees C, 5% CO2). Basal adhesion of normal and SCA neutrophils to FN was not statistically different. Pretreatment of normal neutrophils with either IL-6 (10 100 pg/ml), GCSF (1- 10 ng/ml) or IL-8 (1-100 ng/ml) had no significant effect upon their adhesion to FN. In contrast, SCA neutrophil adhesion to FN was increased significantly following pre-incubation with IL-6, G-CSF and IL-8 (p < 0.01). RANTES (1-100 ng/ml) had no significant effect on either normal or SCA neutrophil adhesion to FN. Flow-cytometric analyses demonstrated that IL-8 (10 ng/ml) significantly augments CD11b (Mac-1 integrin subunit) expression on SCA neutrophils, but not normal neutrophils. IL-6 and G-CSF (10 pg/ml and 10 ng/ml, respectively), however, had no effect on SCA neutrophil adhesion molecule expression. In conclusion, SCA neutrophil adhesion mechanisms may increase in the presence of certain cytokines, in vivo, and this activation may contribute to the physiopathology of sickle cell disease. PMID- 15802893 TI - The incidence of myelofibrosis in essential thrombocythaemia, polycythaemia vera and chronic idiopathic myelofibrosis: a retrospective evaluation of sequential bone marrow biopsies. AB - The incidence of myelofibrosis (MF) among the three major Philadelphia chromosome negative chronic myeloproliferative disorders, i.e. essential thrombocythaemia (ET), polycythaemia vera (PV) and chronic idiopathic myelofibrosis (CIMF), is not well documented since the diagnostic criteria have recently been redefined by the WHO. Therefore we performed a retrospective analysis of follow-up biopsies of 275 patients with ET, PV and CIMF according to the WHO classification of chronic myeloproliferative disorders. In the diagnostic bone marrow biopsies, MF was observed in 57 of the 136 CIMF patients (42%), 4 of the 73 PV patients (5%) and none of the 66 patients with ET. Within a median observation time of 2.9 years, 34 of the 79 patients with CIMF (43%), 13 of the 69 patients with PV (19%) and 1 of the 66 patients with ET (1.5%)--each initially without MF--developed MF regardless of myelosuppressive therapy. PMID- 15802894 TI - CD20-negative pyothorax-associated B cell lymphoma. AB - We describe an 86-year-old male who developed CD20-negative pyothorax-associated B cell lymphoma 64 years after he had suffered from tuberculous pleuritis. Therapy with 8 courses of THP-COP at 2-week intervals was followed by involved field radiotherapy of 30 Gy. Uncertain complete remission was achieved. Thereafter, local recurrence of pyothorax-associated lymphoma (PAL) at the primary site was seen. The patient received salvage radiotherapy of 50 Gy. The patient died of pneumonia during a second uncertain complete remission. The progression-free survival and overall survival of this patient were 10 and 15 months, respectively. When compared with the median survival of 9 months reported in the literature, the adverse effect of CD20 negativity on prognosis may not apply to PAL patients with an occasional aberrant phenotype. PMID- 15802895 TI - Autoimmune disorders in two patients with myelodysplastic syndrome and 5q deletion. AB - Autoimmune diseases occurring concurrently with myelodysplastic syndrome (MDS) with deletion del(5q) including band q31 are very rare and have only been reported twice in the medical literature. We present two additional cases, one patient with del(5q) and trisomy 21 who suffered from rheumatoid arthritis and one patient with isolated del(5q) and autoimmune hemolytic anemia. Both patients had mild leukopenia and severe transfusion-dependent anemia. The rheumatoid arthritis was treated with antirheumatics without additional immunosuppressive medication. Autoimmune hemolytic anemia was controlled with long-term steroid administration. This patient developed additional trisomy 21, 2 years after the initial diagnosis of del(5q). Contrary to previous reports on autoimmune disorders in MDS mentioning improvements of hematological function in response to steroid administration, neither of our patients had a hematological improvement under corticosteroids. PMID- 15802897 TI - The CX3C chemokine fractalkine (CX3CL1) is detectable in serum of B cell chronic lymphocytic leukemia patients with lymph node involvement. PMID- 15802896 TI - Idiopathic systemic capillary leak syndrome misdiagnosed and treated as polycythemia vera. PMID- 15802898 TI - Carnitine therapy is associated with decreased hospital utilization among hemodialysis patients. AB - BACKGROUND/AIMS: Hospitalizations account for 41% of the total cost of end-stage renal disease (ESRD) care. Carnitine deficiency is common among dialysis patients, and some studies have shown improvements in anemia, and cardiac and skeletal muscle function upon administration of L-carnitine. We hypothesized that L-carnitine may be associated with decreased hospital utilization in these patients. METHODS: The Fresenius Medical Care North America dialysis database was used for this retrospective analysis. Adult patients who received carnitine for at least 3 months, and had at least 3 months of pre-carnitine follow-up were included in the study. Hospitalization and hospital day rates were compared before and during carnitine therapy, and with a matched population. RESULTS: Carnitine therapy at a mean dose of 1.5 +/- 0.7 g per administration for an average of 9.7 +/- 5.4 months was associated with a significant reduction in hospital utilization. Patients with cardiovascular disease, defined as hospitalizations for angina, myocardial infarction, arrhythmia, congestive heart failure, cerebral vascular disease or peripheral vascular disease prior to receiving carnitine, and those with anemia and hypoalbuminemia derived the greatest benefit from carnitine therapy. In a multivariate analysis, compared to 3 months prior to the initiation of carnitine, the adjusted relative risk for hospitalization was 11, 11, and 15% lower at 3, 6, and 9 months, respectively. Among patients with cardiovascular disease, the reduction in risk was even more significant (24, 31, and 34% lower at 3, 6, and 9 months, respectively). Similar results were observed with adjusted relative risk for hospital days. CONCLUSION: Administration of L-carnitine to chronic hemodialysis patients is associated with lower hospital utilization. PMID- 15802899 TI - Angiographic characteristics of renal arterial disease over the spectrum of coronary artery disease. AB - BACKGROUND/AIM: The angiographic characteristics of renal artery stenoses (RAS) in patients with coronary artery disease (CAD) have not been yet fully investigated. We sought to evaluate the angiographic characteristics of RAS in patients with CAD. METHODS: The medical records of consecutive patients who underwent coronary angiography in a single public institution over a 12-month period were evaluated. The patients who underwent coincident diagnostic renal angiography to evaluate renal vessels on the basis of clinical criteria and who had at least one-vessel CAD were analyzed. Moderate (50-70%) to severe (70-100%) arterial stenoses were noted as significant angiographic findings. The types of stenosis (ostial, true renal, mixed) and presence and location of calcium were recorded. RESULTS: Angiographically significant RAS were reported in 40 (19.5%) of 205 consecutive patients (mean age 67.1 +/- 12.8 years, mean serum creatinine concentration 2.1 +/- 0.5 mg/dl, mean glomerular filtration rate 52 +/- 13 ml/min) for a total of 55 lesions. The RAS severity was moderate in 30.9% (17/55), severe in 69.1% (38/55), ostial in 27.2% (15/55), true renal in 10.9% (6/55), and mixed in 61.8% (34/55) of the patients. The mean lesion length was 16 +/- 1.8 mm. Patients with > or = 3-vessel CAD had a statistically significantly higher prevalence of mixed calcified RAS (18/24, 75%). Logistic regression analyses revealed > or = 3-vessel CAD (odds ratio 9.917, p = 0.002), age > 65 years (odds ratio 3.817, p = 0.036), and > or = 3 risk factors (odds ratio 2.8, p = 0.048) as independent predictors of RAS. CONCLUSION: RAS in multivessel CAD patients seems to have a peculiar angiographic pattern, such as a higher prevalence of mixed calcified lesions and poststenotic enlargement, that should be taken in account when dealing with RAS. PMID- 15802900 TI - Overweight, obesity and body fat composition in spanish adolescents. The AVENA Study. AB - OBJECTIVE: To describe the prevalence of overweight and obesity in the Spanish adolescent population and its relationship with the socioeconomic status, and to assess their body fat composition and compare these results with previous data from our own country. DESIGN: Cross-sectional multicenter study conducted in five Spanish cities (Granada, Madrid, Murcia, Santander and Zaragoza) in 2000-2002. SUBJECTS: 2,320 adolescents with complete set of anthropometric measurements, 1,192 boys and 1,128 girls. MEASUREMENTS: Body mass index calculated from weight and height measurements, and body fat percentage calculated from skinfold thickness measurements. RESULTS: Overweight + obesity prevalences were 25.69 and 19.13% in boys and girls, respectively. Overweight + obesity prevalence increased in boys from high to medium-low socioeconomic status categories (p = 0.015); meanwhile, there was not a significant effect of socioeconomic status in girls. In males, overweight + obesity prevalence changed from 1985 to 2000-2002 from 13 to 35% and in females from 16 to 32%. The rate of change in overweight + obesity prevalences seems to increase in the last years; from 0.88 (1985 to 1995) to 2.33%/year (1995 to 2000-2002) in males and from 0.5 (1985 to 1995) to 1.83%/year (1995 to 2000-2002) in females. The rate of body fat percentage increase was similar between 1980 and 1995 and between 1995 and 2000-2002: 0.26 and 0.23%/year, respectively, at 13 years of age, and 0.16 and 0.17%/year, respectively, at 14 years of age. CONCLUSION: We observed elevated overweight and obesity prevalences in Spanish adolescents, similar to those observed in other European countries. There is a significant inverse relationship between socioeconomic status and overweight + obesity, but only in boys. The rate of change in overweight prevalence in Spanish adolescents seems to increase, and the rate of increase of body fat percentage seems to be similar as in previous years. PMID- 15802901 TI - Effects of intermittent fasting on serum lipid levels, coagulation status and plasma homocysteine levels. AB - BACKGROUND: During Ramadan, Muslims fast during the daylight hours for a month. The duration of restricted food and beverage intake is approximately 12 h/day which makes Ramadan a unique model of intermittent fasting. Many physiological and psychological changes are observed during Ramadan that are probably due to the changes in eating and sleeping patterns. METHODS: Serum total cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), prothrombin time, activated partial thromboplastin time (aPTT), plasma fibrinogen, D-dimer and homocysteine levels were measured in 24 healthy fasting volunteers (12 females, 12 males) aged 21-35 years. Venous blood samples were taken 1 week before Ramadan, on the 21st day of Ramadan and 20 days after Ramadan. RESULTS: No significant changes were observed on serum total cholesterol, triglycerides and LDL levels. HDL levels were significantly elevated during Ramadan (p < 0.001) and 20 days after Ramadan (p < 0.05). Prothrombin time, aPTT, fibrinogen and D-dimer levels were in the physiologic limits in all samples but D-dimer levels were significantly low at the end of Ramadan in comparison to pre- and post-fasting levels (p < 0.001). Homocysteine levels, being still in reference ranges, were low during Ramadan (p < 0.05) and reached the pre-fasting levels after Ramadan. CONCLUSION: Our results demonstrate that intermittent fasting led to some beneficial changes in serum HDL and plasma homocysteine levels, and the coagulation status. These changes may be due to omitting at least one meal when the body was particularly metabolically active and possibly had a low blood viscosity level at the same time. We conclude that intermittent fasting may have beneficial effects on hemostatic risk markers for cardiovascular diseases. PMID- 15802902 TI - Relationships between IGF-I and weight Z score, BMI, tricipital skin-fold thickness, type of feeding in healthy infants in the first 5 months of life. AB - AIM: To determine the concentration of insulin-like growth factor-I (IGF-I) in exclusively breast-fed (BF) and formula-fed (FF) infants in the first 5 months of life and to study the relationship between the IGF-I level and Z score for weight, body mass index (BMI) and tricipital skin-fold thickness. METHODS: We performed a cross-sectional study in order to evaluate serum IGF-I in 97 age matched healthy term infants: 50 FF and 47 BF. RESULTS: FF infants showed higher values of IGF-I (38.9 +/- 12 ng/ml) when compared to BF infants (26.7 +/- 11.6 ng/ml; p < 0.05). A positive correlation was found between IGF-I values and Z score for weight (p < 0.001), BMI (p = 0.014), tricipital skin-fold thickness (p = 0.043) and age (p = 0.02). CONCLUSION: These results show that the IGF-I serum concentration is higher in FF infants. The observed association between IGF-I and Z score for weight, BMI, tricipital skin-fold thickness confirms the effect of different infant feeding on body size. PMID- 15802903 TI - Comparisons of serum somatotropin, 3,5,3'-triiodothyronine, thyroxine, total protein and free fatty acid levels in newborn Sakiz lambs separated from or suckling their dams. AB - OBJECTIVE: To determine the effects on serum somatotropin, 3,5,3' triiodothyronine (T(3)), thyroxine (T(4)), free fatty acids (FFAs) and total protein levels of different feedings and age in Sakiz sheep that have a high twin bearing rate supported by estrous synchronization. METHODS: 20 newborn lambs were used in the study. Lambs were divided into 2 equal groups. The lambs in 1 group were separated from their dams following parturition, and those in other group were kept together with their dams. Separated lambs were fed commercial cow's milk for 2.5 weeks. After that, they were fed a milk substitute, hay and concentrated supplement for 2 months, and from the end of the 2 months they were fed hay and concentrated supplement. The lambs in other group were kept together with their dams only during the day and in addition were fed hay and concentrated supplement. After 2 months, they were maintained feeding only on hay and concentrated supplement. Blood samples were taken from vena jugularis of lambs at 12 and 24-48 h, and 14, 28, 42 and 56-90 days after parturition. Serum samples were analyzed for somatotropin by enzyme immunoassay, for T(3) and T(4) by radioimmunoassay, and for total protein and FFA by a spectrophotometric method. RESULTS: Serum somatotropin levels were insignificantly different between the lambs with and without their dams. Serum T(3) levels were generally lower in the separated lambs than those in other group. Serum T(4) levels were significantly lower in the separated lambs 48 h and 90 days after parturition than those in other group. Serum FFAs were insignificantly different between 2 groups. Serum total protein concentrations were lower in the separated lambs than lambs kept together with their dams (significantly at 12 and 48 h and 14 days). CONCLUSION: Ingestion of colostrum and dam milk markedly affected serum total protein levels, but body weight and serum somatotropin levels were insignificantly affected. Also serum T(3) and T(4) levels markedly decreased with advancing age. Serum FFA levels were not affected by the trial. PMID- 15802904 TI - A randomized clinical trial with oral Immunonutrition (omega3-enhanced formula vs. arginine-enhanced formula) in ambulatory head and neck cancer patients. AB - OBJECTIVE: The aim of our study was to investigate whether oral ambulatory nutrition of head and neck cancer patients, using an omega3 fatty acid-enhanced diet (low ratio omega6/omega3 fatty acids) versus an arginine-enhanced diet, could improve nutritional variables as well as clinical outcome, postoperative infectious and wound complications. RESEARCH METHODS: A population of 73 ambulatory postsurgical patients with oral and laryngeal cancer were enrolled. At discharge from hospital the postsurgical head and neck cancer patients were asked to consume two units per day of either a specially designed omega3 fatty acid enhanced supplement (group 1) or an arginine-enhanced supplement (group 2) for a 12-week period. RESULTS: No significant intergroup differences in the trend of the three serum proteins and lymphocytes were detected. Differences were detected in weight (group 1: 65.5 +/- 11.5 kg vs. 70.4 +/- 11.1 kg; p < 0.05) with a significant increase in fat mass in group 1 (15.4 +/- 6.6 vs. 18.1 +/- 8.4 kg; p < 0.05) and in tricipital skinfold. The postoperative infectious complications were similar in both groups (0 in group 1 and 8.57% in group 2; nonsignificant). No local complications were detected in the surgical wound. Gastrointestinal tolerance (diarrhea and vomiting episodes) of both formulas was good. CONCLUSIONS: At the dose taken, the omega3-enhanced formula improved fat mass and proteins in ambulatory postoperative head and neck cancer patients. The arginine enhanced formula improved proteins. Further studies are required to examine the potential role of immune-enhanced supplements. PMID- 15802905 TI - Adult hypophosphatasia and a low level of red blood cell thiamine pyrophosphate. AB - OBJECTIVE: To report the first adult case of hypophosphatasia and absence of intestinal alkaline phosphatase (ALP) isoenzymes associated with a low level of red blood cell thiamine pyrophosphate. METHODS: We describe the clinical manifestation and laboratory findings in our patient and discuss underlying factors that potentially contribute to her condition. RESULTS: A 33-year-old Vietnamese-American female was referred for a long history of low levels of serum ALP and absence of intestinal ALP isoenzyme. She had a normal level of urinary phosphoethanolamine and a high level of plasma pyridoxal-5p-phosphate. She was found to have a low level of red blood cell thiamine pyrophosphate. CONCLUSION: Our adult case of hypophosphatasia presented with an absence of intestinal ALP isoenzyme that might result in decreasing the absorption of thiamine in the intestinal tract. Therefore, the thiamine level should be considered in hypophosphatasia with absence of intestinal ALP isoenzyme. PMID- 15802906 TI - Assessment of body composition and breast milk volume in lactating mothers in pastoral communities in Pokot, Kenya, using deuterium oxide. AB - BACKGROUND: In sub-Saharan Africa, the practice of breast-feeding infants is common. Records documenting the intake of breast milk amongst infants are limited. This study evaluated the association between maternal body composition and the intake of breast milk in infants from the pastoral communities within Pokot, Kenya. METHODS: The study was conducted in 10 lactating mothers who were participating in a longitudinal study aimed at determining maternal body composition, iron stores and vitamin A status during the third trimester pregnancy and four months after they had given birth. Maternal and infant anthropometric measurements were made, and maternal blood samples were taken to determine serum retinol and ferritin levels. Infant milk intake and maternal fat free mass (FFM) and percent body fat (% BF) were measured using 'the dose to the mother method'. A measured deuterium oxide ((2)H(2)O) dose was given to the mother. Urine and breast milk from the mother, and saliva samples from the infant, were collected on days 1, 8 and 14 after dosing. RESULTS: The mean (+/- SD) maternal mid upper arm circumference (MUAC) and body mass index (BMI) were 21.8 (0.9) cm and 18.6 (1.0) kg/height (m(2)), respectively. Infant weight and weight/age Z score were 4.956 (0.874) kg and -1.750 (0.77), respectively. Throughout the study, the infants gained 20 (4) g/day in body weight and had a milk intake of 555 (22) ml/day. The energy intake of the infant was 1,602 (148) kJ/day and was lower (p < 0.05) than the 2,404 (423) kJ/day estimated requirement by the FAO/WHO/UNU. The maternal FFM, %BF, Hb, Hct, ferritin and retinol were 32.8 (3.1) kg, 17.24 (7.0), 11.5 (1.3) g/dl, 33.9 (4.9), 16.2 (0.1) microg/l and 0.894 (0.16) micromol/l, respectively. Infant milk intake was significantly and positively correlated to maternal pregnancy triceps (r = 0.679) p < 0.05) and pregnancy MUAC (r = 0.725) p < 0.05). Maternal pregnancy MUAC was an important predictor of infant breast milk intake. CONCLUSION: Data on volume of breast milk consumed by the infants suggests, at least for this group of infants, that adequate growth may not be achieved. There is a possibility that lactating mothers practicing exclusive breast-feeding and living under harsh conditions may experience periods of low breast milk volume. Body composition and biochemical findings among this group of Pokot mothers indicate dietary inadequacies that require nutritional intervention. PMID- 15802908 TI - Mal-positioned umbilical venous catheter causing liver abscess in a preterm infant. AB - Neonatal liver abscess is uncommon, carries a high mortality and is difficult to diagnose. We report an unusual case of liver abscess in a preterm infant presenting with abdominal distension and suspected gastrointestinal perforation, rather than the more usual features of fever, hepatomegaly, abdominal tenderness, right-sided pleural effusion, and leukocytosis. We discuss current treatments for neonatal liver abscess and argue that in view of the high mortality and difficulty in diagnosis, prevention should be the primary objective. We believe that mal-positioning the umbilical venous catheter in the liver substantially increases the life-threatening risk of this complication, and advocate extreme care in the placement and use of these catheters. PMID- 15802907 TI - Effect of green tea decoction on long-term iron, zinc and selenium status of rats. AB - AIMS: The objective was to examine the effect of green tea decoction given at two different concentrations on the long-term (6 weeks) iron, zinc and selenium status of rats. METHODS: During the experimental period, the rats were given ad libitum a basic diet + ultra pure water (control group), a basic diet + green tea decoction prepared from 50 g/l (tea 50 group), or a basic diet + green tea decoction prepared from 100 g/l (tea 100 group). The zinc and iron status was evaluated by determining their concentrations in the serum, blood precipitate, liver, spleen, femur, heart and kidney. Selenium status was evaluated by the serum selenium concentration and whole blood glutathione peroxidase activity. RESULTS: Green tea decoction significantly reduced serum iron by 26% in the tea groups (p < 0.01). The blood precipitate of iron was significantly decreased by 25 and 41% in the tea 50 and tea 100 groups (p < 0.01), respectively. The reserve of iron stored in the liver, spleen and femur was significantly reduced in the tea 100 group by 32% (p < 0.02), 20% (p < 0.04) and 35% (p < 0.005), respectively. Moreover, the two concentrations of green tea significantly decreased the reserve of iron stored in the kidney (p < 0.005) and heart (p < 0.02). In contrast with its effects on iron status, green tea decoction significantly increased the serum zinc in the tea 100 group by 24% (p < 0.001). It also increased the blood precipitate of zinc by 50 (p < 0.01) and 75% (p < 0.0001) in tea 50 and tea 100 groups, respectively. In the kidney, heart and femur, zinc significantly increased in the tea groups dependent on the tea dose. Similarly, the high concentration of green tea decoction significantly increased the serum selenium concentration by 16% (p < 0.004). In addition, both concentrations of green tea decoction significantly increased the whole blood glutathione peroxidase activity by 102 and 130% (p < 0.01). CONCLUSION: Green tea decoction reduced the iron status and improved the zinc and selenium status of rats. These effects may constitute another beneficial effect of the green tea decoction which could play an important role in the antioxidant processes. PMID- 15802909 TI - Anosognosia in very mild Alzheimer's disease but not in mild cognitive impairment. AB - OBJECTIVE: To study awareness of cognitive dysfunction in patients with very mild Alzheimer's disease (AD) and subjects with mild cognitive impairment (MCI). METHODS: A complaint interview covering 13 cognitive domains was administered to 82 AD and 79 MCI patients and their caregivers. The patient groups were comparable according to age and education, and Mini Mental State Examination (MMSE) scores were > or =24 in all cases. The discrepancy between the patients' and caregivers' estimations of impairments was taken as a measure of anosognosia. RESULTS: Self-reports of cognitive difficulties were comparable for AD and MCI patients. However, while in comparison to caregivers MCI patients reported significantly more cognitive impairment (p < 0.05), AD patients complained significantly less cognitive dysfunctions (p < 0.001). CONCLUSIONS: While most MCI patients tend to overestimate cognitive deficits when compared to their caregiver's assessment, AD patients in early stages of disease underestimate cognitive dysfunctions. Anosognosia can thus be regarded as a characteristic symptom at a stage of very mild AD (MMSE > or =24) but not MCI. Accordingly, medical history even in mildly affected patients should always include information from both patient and caregiver. PMID- 15802910 TI - The k variant of the butyrylcholinesterase gene is associated with reduced phosphorylation of tau in dementia patients. AB - Accumulating evidence suggests that butyrylcholinesterase (BuChE) plays an important role in the progression of cognitive deficits and Alzheimer-type pathology in dementia patients. We examined the relationship between the K variant of BuChE and the severity of deposits of amyloid (Abeta(1-42)) and phosphorylated tau in the temporal cortex (BA36) of 30 prospectively studied autopsy-diagnosed dementia (Alzheimer's disease and dementia with Lewy bodies) patients. There was 42% less phosphorylated tau in BA36 in cases with > or =1 K compared with those with wild-type BuChE alleles (t = 2.2, p = 0.039), but no difference in the extent of Abeta(1-42) deposition. BuChE may play this role in the phosphorylation of tau, relevant to therapeutic inhibition of the enzyme. PMID- 15802911 TI - Peripheral electrical stimulation in Alzheimer's disease. AB - In a number of studies, peripheral electrical nerve stimulation has been applied to Alzheimer's disease (AD) patients who lived in a nursing home. Improvements were observed in memory, verbal fluency, affective behavior, activities of daily living and on the rest-activity rhythm and pupillary light reflex. The aim of the present, randomized, placebo-controlled, parallel-group clinical trial was to examine the effects of electrical stimulation on cognition and behavior in AD patients who still live at home. Repeated measures analyses of variance revealed no effects of the intervention in the verum group (n = 32) compared with the placebo group (n = 30) on any of the cognitive and behavioral outcome measures. However, the majority of the patients and the caregivers evaluated the treatment procedure positively, and applying the daily treatment at home caused minimal burden. The lack of treatment effects calls for reconsideration of electrical stimulation as a symptomatic treatment in AD. PMID- 15802912 TI - Mood and cognitive function in elderly diabetic patients living in care facilities. AB - Community studies found deficits of cognitive function in elderly persons with diabetes. The aim of the present study was to compare diabetes and other age related diseases among residents of senior citizens' care facilities. Participants were 130 residents with and 130 without a diagnosis of diabetes aged between 65 and 98 years. Measures were neuropsychological tests designed for this age group, questionnaires of mood, quality of life and daily activities and level of glycosylated hemoglobin (HbA(1c)) as an index of metabolic control. Only verbal memory was significantly more impaired in residents with than without diabetes. Otherwise, both groups showed overall impairment of cognitive function and mood. Unexpectedly, a slightly elevated HbA(1c) level (6-8%) was associated with an increased level of cognitive and everyday functioning. In old age, diabetic patients might benefit from a slightly higher level of blood glucose than has hitherto been assumed. PMID- 15802913 TI - Follow-up investigations of tau protein and S-100B levels in cerebrospinal fluid of patients with Creutzfeldt-Jakob disease. AB - BACKGROUND: S-100B and tau protein have a high differential diagnostic potential for the diagnosis of Creutzfeldt-Jakob disease (CJD). So far there has been only limited information available about the dynamics of these parameters in the cerebrospinal fluid (CSF). However, there is a special interest in finding biochemical markers to monitor disease progression for differential diagnosis and treatment. PATIENTS AND METHODS: We analyzed CSF of 45 patients with CJD and of 45 patients with other neurological diseases for tau protein and S-100B in a follow-up setting. All diagnoses of CJD were later neuropathologically verified. A ratio between tau protein differences and the time between lumbar puncture was calculated. The same was done for S-100B. RESULTS: Tau protein levels of 34 cases were above the cut-off level for CJD (>1,300 pg/ml) in the first CSF sample. In 7 of 11 patients with lower tau levels in the first CSF sample, tau levels rose. The above-mentioned ratio was significantly higher in the CJD group than in the group with other neurological diseases. Similar results were obtained for S-100B. CONCLUSION: We conclude that follow-up investigations and calculation of ratios is a useful tool in the differential diagnosis of CJD. Variations in this pattern were observed in single cases. PMID- 15802914 TI - Mild cognitive impairment: an operational definition and its conversion rate to Alzheimer's disease. AB - OBJECTIVE: Because of discrepant findings regarding the accuracy of mild cognitive impairment (MCI) in predicting Alzheimer's disease (AD), further study of this construct and conversion rates is essential before use in clinical settings. We aimed to develop an operational definition of MCI consistent with criteria proposed by the Mayo Alzheimer's Disease Center, and to examine its conversion rate to AD. METHODS: Patients were identified from an inception cohort of patients with at least a 3-month history of memory problems, and referred to a 2-year university teaching hospital investigation by primary care physicians. We classified 161 nondemented patients at baseline using MCI criteria. Diagnostic workups were completed annually, and patients were classified as meeting criteria for AD or showing no evidence of dementia after 1 and 2 years. RESULTS: Of 161 patients, 35% met MCI criteria at baseline. Conversion rates to AD were 41% after 1 year, and 64% after 2 years. Logistic regression analyses to examine predictive accuracy of MCI after 1 and 2 years, with age and education as covariates, were significant (p < 0.0001). After 1 year, MCI showed an optimal sensitivity of 91% and specificity of 79%, and after 2 years, these values were 88 and 83%, respectively. CONCLUSIONS: MCI is an accurate predictor of AD over 1 and 2 years in patients referred by their primary care physicians. Discrepancies in conversion rates may be due to the manner in which patients are recruited to studies as well as the use of different measures to operationalize the construct. PMID- 15802915 TI - Neural correlates of impaired performance on the clock drawing test in Alzheimer's disease. AB - To identify the neural correlates for impaired performance on the clock drawing test (CDT) in patients with Alzheimer's disease (AD), we examined the relationship between the CDT performances and the regional cerebral blood flow (rCBF) in 100 AD patients. The patients were equally divided into a mildly impaired CDT group, a severely impaired CDT group, and two normal CDT groups, with age and dementia severity matched. Between-group comparisons revealed that rCBF reduction in the posterolateral region of the left temporal lobe was consistently associated with mild to severe impairment of the CDT in AD. Correlation analysis also showed that the rCBF in the left posterolateral temporal cortex was linearly correlated with CDT performance. The CDT scores in AD were significantly improved for the copy condition relative to the drawing-to command condition. These findings suggest that CDT performance has a close relationship with the left posterior temporal function, and that semantic memory deficit may at least partly contribute to impaired CDT performance in AD. PMID- 15802916 TI - A confidence set inference procedure for gene mapping using markers with incomplete polymorphism. AB - A recent approach for gene mapping based on confidence set inference (CSI) promises several advantages, including avoidance of corrections for multiple tests, availability of confidence intervals with known statistical properties, and sufficient localizations of disease genes. This paper proposes an extended CSI procedure that can handle markers with incomplete polymorphism, thereby increasing the applicability of the set of CSI methods in practical situations. Simulation studies show that the new procedure retains the main advantages of the original CSI. Although it generally requires more data to achieve a similar power, this increase is moderate for markers with 80% heterozygosity or higher. We also investigate the effects of relative risk estimates and disease models. Our analyses show that perturbation from actual relative risks or multilocus disease models generally leads to reduction in power or inflation in type I error, as expected. Nevertheless, for certain classes of two-locus disease models, CSI can still perform well, with reasonably high actual coverage probabilities for at least one of the disease loci. Application of CSI to the data provided by the Genetic Analysis Workshop 13 yields encouraging results, as they compare favorably to those obtained from GENEHUNTER using its NPL sib-pair method. PMID- 15802917 TI - On averaging power for genetic association and linkage studies. AB - A power calculation is crucial in planning genetic studies. In genetic association studies, the power is often calculated using the expected number of individuals with each genotype calculated from an assumed allele frequency under Hardy-Weinberg equilibrium. Since the allele frequency is often unknown, the number of individuals with each genotype is random and so a power calculation assuming a known allele frequency may be incorrect. Ambrosius et al. recently showed that the power ignoring this randomness may lead to studies with insufficient power and proposed averaging the power due to the randomness. We extend the method of averaging power in two directions. First, for testing association in case-control studies, we use the Cochran-Armitage trend test and find that the time needed for calculating the averaged power is much reduced compared to the chi-square test with two degrees of freedom studied by Ambrosius et al. A real study is used for illustration of the method. Second, we extend the method to linkage analysis, where the number of identical-by-descent alleles shared by siblings is random. The distribution of identical-by-descent numbers depends on the underlying genetic model rather than the allele frequency. The robust test for linkage analysis is also examined using the averaged powers. We also recommend a sensitivity analysis when the true allele frequency or the number of identical-by-descent alleles is unknown. PMID- 15802918 TI - A note on linkage analysis with affected sib triplets. AB - Previously, it has been shown for affected sib pairs that the mean test is the uniformly (in theta) most powerful test in case of a multiplicative mode of inheritance and that the mean test is equivalent to parametric linkage analysis calculated under an assumed multiplicative mode of inheritance. Here, these two results are extended to samples consisting of affected sib triplets. For affected sib quadruplets, however, it is shown that these results are no longer valid. PMID- 15802919 TI - Parametric approach to genomic imprinting analysis with applications to Angelman's syndrome. AB - Genomic imprinting is a mechanism by which only one copy of a gene pair is expressed, and this expression is determined by the parental origin of the copy. The deregulation of imprinted genes has been implicated in a number of human diseases. The Imprinted Gene Catalogue now has more than 200 genes listed, and estimates based on mouse models suggest many more may exist in humans. Therefore, the development of methods to identify such genes is important. In this communication, we present a parametric model-based approach to analyzing arbitrary-sized pedigree data for genomic imprinting. We have modified widely used LINKAGE program to incorporate our proposed approach. In addition, our approach allows for the use of sex-specific recombinations in the analysis, which is of particular importance in a genome-wide analysis for imprinted genes. We compared our imprinting analysis approach to that implemented in the GENEHUNTER IMPRINT program using simulation studies as well as by analyzing causal genes in Angelman's syndrome families, which are known to be imprinted. These analyses showed that the proposed approach is very powerful for detecting imprinted genes in large pedigrees. PMID- 15802920 TI - Common genetic and environmental effects on lipid phenotypes: the HERITAGE family study. AB - OBJECTIVE: Despite the well known genetic component influencing plasma lipid lipoprotein levels and the observed correlations among these traits, little is known about pleiotropic heritable determinants among them. Our aim is to investigate pair-wise polygenic and environmental correlations among lipid lipoprotein levels at baseline and in response to regular exercise in Whites and Blacks. METHODS: Common pair-wise genetic and environmental correlations among levels of total cholesterol (TC), LDL-C, ApoB, HDL-C (also HDL2-C and HDL3-C), triglycerides (TG, HDL-TG and LDL-TG) and ApoA-1 were investigated at baseline and again after a 20-week endurance exercise program using a variance-components decomposition. RESULTS: With a few exceptions, all lipid phenotypes were heritable at baseline and for training responses in Blacks and Whites. Strong to high genetic and environmental correlations (0.4 < rho(g) < 0.7) were observed for the majority of the baseline pair-wise traits. For training responses, many of the same patterns were noted, although fewer genetic correlations were significant as compared to the baseline results. CONCLUSIONS: Results suggest that the observed phenotypic correlations among many of these traits may be due to in part to pleiotropic genes, in particular between LDL-C and ApoB and between TG and HDL-C. This shared genetic architecture should be considered in follow-up gene finding studies. PMID- 15802921 TI - Maximum likelihood haplotyping for general pedigrees. AB - Haplotype data is valuable in mapping disease-susceptibility genes in the study of Mendelian and complex diseases. We present algorithms for inferring a most likely haplotype configuration for general pedigrees, implemented in the newest version of the genetic linkage analysis system SUPERLINK. In SUPERLINK, genetic linkage analysis problems are represented internally using Bayesian networks. The use of Bayesian networks enables efficient maximum likelihood haplotyping for more complex pedigrees than was previously possible. Furthermore, to support efficient haplotyping for larger pedigrees, we have also incorporated a novel algorithm for determining a better elimination order for the variables of the Bayesian network. The presented optimization algorithm also improves likelihood computations. We present experimental results for the new algorithms on a variety of real and semiartificial data sets, and use our software to evaluate MCMC approximations for haplotyping. PMID- 15802922 TI - Follicular cell-derived thyroid cancer in children. AB - Thyroid carcinoma is a rare disease in childhood. The only known causative factor is radiation exposure during childhood. Most cases can be cured by surgery and eventually radioiodine. The aim of initial treatment should be the total removal of neoplastic foci with a minimal morbidity. Some cancer-related deaths have been reported decades after initial treatment. PMID- 15802923 TI - The effect of different diagnostic criteria on the prevalence and incidence of post-stroke dementia. AB - There is little agreement about the prevalence and incidence of vascular dementia (VaD) mainly because investigators have used different diagnostic criteria. The aim of this study was to examine the influence of different diagnostic criteria on the prevalence and cumulative incidence of VaD in first-ever stroke patients (n = 194) clinically evaluated at 1, 6, 12, and 24 months after stroke. Post stroke VaD was diagnosed using the DSM-III, DSM-III-R, DSM-IV, ICD-10-NA, NINDS AIREN, and ADDTC criteria. The prevalence of dementia was highest at 1 month; ranging from 11.3% with the NINDS-AIREN to 20.1% with the ICD-10-NA. The incidence was highest at 6 months, ranging from 2.6% with the ADDTC to 5.2% with the ICD-10-NA. Agreement among diagnostic criteria was high, with the exception of the ADDTC. In conclusion, both the prevalence and incidence of dementia are highest directly after stroke, but exact rates are influenced by the diagnostic criteria used. PMID- 15802924 TI - The use of the 12-item short-form health status instrument in a longitudinal study of patients with stroke and transient ischaemic attack. AB - The 36-item short-form health survey (SF-36) is one of the most commonly used health status instruments in patients with cerebrovascular disease. However, responsiveness to change in health-related quality of life (HRQoL) has not yet been assessed for the SF-36 and its shortened version, the SF-12. The main objective of the present study was to determine responsiveness to change of the SF-12 in patients with cerebrovascular disease. Patients with stroke/transient ischaemic attack (TIA) were included at admission to one of four participating hospitals. HRQoL was assessed with the Physical (PCS-12) and Mental (MCS-12) Component Summary scales at baseline (referring to the status prior to the event) and after 12 months. Responsiveness to change was determined with the standardized response mean (SRM) and classified as small (SRM 0.20-0.49), moderate (0.50-0.79) or large (> or =0.80). A total of 558 patients were included [55% men, mean age 65 (SD, 13) years; 45% women, mean age 69 (SD, 14) years]. Indications for admission were stroke (74%) and TIA (26%). In patients with stroke, SRMs were small for the PCS-12 [SRM 0.49; absolute change -5.9 (SD, 12)] and moderate for the MCS-12 [SRM 0.52; absolute change -6.6 (SD, 13)]. In patients with TIA, SRMs were below 0.2 for the PCS-12 [absolute change -0.7 (SD, 11)] and small for the MCS-12 [SRM 0.34; absolute change -3.7 (SD, 11)]. SRMs increased with stroke severity as indicated by the NIHSS score. The SF-12 summary scales showed a small to moderate responsiveness to change in patients after stroke. Responsiveness to change was higher in patients with increased symptom severity. PMID- 15802925 TI - Case-control study of nutritional antioxidant intake in essential tremor. AB - The theory that oxidative stress is involved in the pathogenesis of neurodegenerative diseases has received considerable attention and studies have linked these diseases to the diminished use of antioxidant vitamins (vitamins E and C) and other dietary antioxidants. Essential tremor (ET) is a chronic, progressive disease. One possible disease mechanism is neurodegenerative. Whether nutritional antioxidant use differs between ET cases and controls is not known. Using a case-control design, we conducted detailed dietary assessments and tested the hypothesis that diminished use of nutritional antioxidants is associated with ET. Data on diet were collected on 156 ET cases and 220 controls using a semi quantitative food-frequency questionnaire. There was no evidence that current nutritional antioxidant exposure differs in ET cases and controls. This does not exclude the possibility that nutritional antioxidant exposure was lower in ET cases prior to their disease onset. PMID- 15802926 TI - Higher burden of hemorrhagic stroke among women. An autopsy-based study in Sao Paulo, Brazil. AB - The aim of this study was to verify the gender difference in sudden cardiovascular death, mainly comparing brain infarction and hemorrhagic stroke. We analyzed 970 autopsy cases from a total of 3,802 nonviolent deaths (448 considered as sudden deaths, 296 undetermined, and 226 non-sudden deaths) including patients aged 30-69 years. All cardiovascular diseases were responsible for 69% of sudden deaths, and all types of stroke for only 14%. The proportions of sudden death for all cardiovascular diseases, coronary heart disease and ischemic stroke deaths were similar for both sexes. However, sudden deaths due to all types of stroke (women 20.0%; men 11.1%; p = 0.012) and hemorrhagic stroke (women 15.6%; men 7.9%; p = 0.019) were significantly more frequent among women when compared to men, the main cause being subarachnoid hemorrhage (women 5.6%; men 1.0%; p = 0.011). Hemorrhagic stroke deaths were usually sudden deaths among women. PMID- 15802928 TI - Association of regular aspirin use and breast cancer risk. AB - Of the limited number of epidemiological investigations on aspirin (and other nonsteroidal anti-inflammatory drugs) and breast cancer, the majority observe a protective role, yet only a few report dose-response effects for frequency or duration of use. We studied aspirin use among 1,478 breast cancer patients diagnosed from 1982 to 1998, and 3,383 cancer-free hospital controls at the Roswell Park Cancer Institute. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using unconditional logistic regression. Compared to never use,both regular (> or =1 tablet per week for > or =1 year) and occasional use were inversely associated with breast cancer (adjusted OR = 0.84, 95% CI 0.64-0.97; adjusted OR = 0.80, 95% CI 0.67-0.96, respectively). Among regular users, an inverse trend was found for number of tablets consumed per week (1, 2-6, or > or =7) with corresponding ORs of 0.95, 0.80, and 0.74 (P(trend) = 0.01). Daily use spanning 10 or more years was associated with a more pronounced reduction in risk (P(trend) = 0.005). Our findings corroborate the growing body of observational evidence that regular aspirin use may be associated with reduced risk of breast cancer. PMID- 15802927 TI - Beta-endorphin ameliorates synovial cell hyperfunction in the collagen-induced arthritis rat model by specific downregulation of NF-kappa B activity. AB - OBJECTIVES: To observe the multiple immunoregulating effects of beta-endorphin (beta-END) on synovium cells of collagen-induced arthritis (CIA) in rats and to determine whether the regulation involves the transcriptional factor-kappaB (NF kappaB) signal pathway. METHODS: CIA was induced in female Wistar rats by immunization with native bovine type-II collagen emulsified with complete Freund's adjuvant. Synovial cells in the knees of the CIA rats were cultivated, and the effects of beta-END, beta-END receptor inhibitor (naloxone, Nal) in proliferation and apoptosis of the synovial cells were assayed by 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, flow cytometry, and DNA integrity, respectively. The effects of beta END and Nal on mRNA expression of several cytokines in the synovial cells, including tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-6, regulated upon activation normal T-cell expressed and secreted (RANTES), inducible nitric oxide synthase (iNOS), matrix metalloproteinase-2 (MMP-2) and MMP-9 were estimated by quantitative reverse transcription-polymerase chain reaction. Effects of beta-END and Nal on NF-kappaB activity were analyzed using luciferase gene reporter assays. The effects of beta-END and Nal on p65NF-kappaB expression of the synovial cells were examined using Western blot. RESULTS: 75% of the rats were demonstrated to have established the CIA model successfully. beta-END was shown to exert multiple effects on synovial cells of CIA rats including decreased proliferation, induced apoptosis, and downregulation of TNF alpha, IL-1beta, IL-6, RANTES, iNOS, MMP-2 and MMP-9 mRNA expression. beta-END seemed to play an immunoregulating role by downregulating the activity and expression of NF-kappaB. It was found that the beta-END receptor blockage could counteract all the effects. CONCLUSIONS: beta-END ameliorates synovial cell functions of CIA rats through binding with receptors and downregulating the NF kappaB signal pathway. This suggests that beta-END, by blocking the activity and expression of NF-kappaB, is a potential anti-inflammatory and anti-rheumatic agent against CIA. PMID- 15802929 TI - Basal cell carcinomas of the eyelids. AB - Basal cell carcinomas (BCC) are the more frequent malignant tumors seen in France as in other western countries. They represent 20% of eyelid tumors and 90% of eyelid malignancies. Due to their local growth, problems may arise when treating BCC, and curative exeresis must be the preferred choice each time it is possible. BCC of the eyelids have a high risk of recurrence. Recurrences are more aggressive, infiltrative and destructive and have a considerably poorer rate of cure than primary tumors. Eyelid reconstructions can entail use of complex methods which should only be carried out by a trained ophthalmologist who is also able to treat any associated age-related ocular pathologies. BCC is the most common cause leading to eyelid reconstructive surgery; a surgery which has a triple objective: tumor removal, functionality and an esthetic outcome. PMID- 15802930 TI - Visual results after primary intraocular lens implantation or contact lens correction for aphakia in the first year of age. AB - PURPOSE: The optimal role of intraocular lenses (IOLs) in infants remains a controversial topic. Some ophthalmologists advocate correction with a contact lens (CL), whereas others recommend an IOL correction. Our study compared visual acuity, ocular alignment, retreatment rate and binocular vision outcomes in children treated with these two methods at our clinic. METHODS: This study included 41 children with unilateral congenital cataract who underwent cataract surgery with posterior capsulorhexis and anterior vitrectomy, coupled with (IOL group, n=18) or without (CL group, n=23) primary IOL implantation. All infants underwent the first surgery during the first 12 months of their life and they were operated on in the period from 1994 to 1999. The mean age at surgery was 3.11+/-2.65 months (range: 28 days to 11 months). All patients were prescribed the same half-time reduced occlusion therapy. Good cooperation of the parents and good compliance with patching were the necessary conditions to include a patient in the study. Between January and February 2003, the final visual acuity and binocular vision outcomes were examined. RESULTS: The mean final visual acuity (logarithm of the minimum angle of resolution) of the operated eye was 0.43+/ 0.33 for the IOL group and 0.58+/-0.39 for the CL group (p=0.14). The mean interocular difference in visual acuity was 0.22+/-0.29 for the IOL group and 0.56+/-0.31 for the CL group (p=0.042). The reoperation rate was 78% in the IOL group compared with 35% in the CL group (p=0.017). Esotropia or exotropia of more than 8 prism diopters were present in 55% of children (10/18) in the IOL group compared with 83% of children (19/23) in the CL group (p=0.039). CONCLUSIONS: We suggest that correction of aphakia after unilateral congenital cataract surgery with primary IOL implantation results in improved visual acuity, improved binocular vision outcome and less occurrence of strabismus, but a higher rate of complications requiring reoperation. Further studies with a larger pediatric patient group are necessary to confirm the optimal treatment of aphakia after unilateral congenital cataract extraction. PMID- 15802931 TI - Optical coherence tomography measurement of the retinal nerve fiber layer in normal and juvenile glaucomatous eyes. AB - The aim of this study was to quantitatively assess and compare the thickness of the retinal nerve fiber layer (RNFL) in normal and glaucomatous eyes of children using the optical coherence tomograph. The mean RNFL thickness of normal eyes (n=26) was compared with that of glaucomatous eyes (n=26). The eyes were classified into diagnostic groups based on conventional ophthalmological physical examination, Humphrey 30-2 visual fields, stereoscopic optic nerve head photography, and optical coherence tomography. The mean RNFL was significantly thinner in glaucomatous eyes than in normal eyes: 95+/-26.3 and 132+/-24.5 microm, respectively. More specifically, the RNFL was significantly thinner in glaucomatous eyes than in normal eyes in the inferior quadrant: 87+/-23.5 and 122+/-24.2 microm, respectively. The mean and inferior quadrant RFNL thicknesses as measured by the optical coherence tomograph showed a statistically significant correlation with glaucoma. Optical coherence tomography may contribute to tracking of juvenile glaucoma progression. PMID- 15802932 TI - Optical coherence tomographic assessment of diabetic macular edema: comparison with fluorescein angiographic and clinical findings. AB - PURPOSE: To compare the optical coherence tomographic (OCT) features with clinical and fluorescein angiographic (FA) findings in patients with diabetic retinopathy. METHODS: In a retrospective study ophthalmologic examination together with FA and OCT images were obtained from 195 eyes of 110 patients with different stages of diabetic retinopathy and OCT images were obtained from 40 eyes of 20 control subjects. Fluorescein leakage characteristics were organized into five groups: no leakage (1), focal (2), diffuse (3), combined focal+diffuse leakage (4) and cystoid (5). The Pearson correlation test was used to test the correlation between visual acuity and central foveal thickness and ANOVA was used for the statistical comparison between the groups. RESULTS: The OCT images demonstrated retinal swelling in 66.1% of eyes, cystoid macular edema (CME) in 11.8% of eyes, serous foveal detachment + swelling in 6.2% of eyes, serous foveal detachment+swelling+CME in 3.6% of eyes and normal foveal structure in 12.3% of eyes. The best-corrected visual acuity was significantly correlated with central foveal thickness (r:-0.528, p<0.01). There was 77% agreement between clinical examination and OCT results. CME was detected with OCT in 15.4% of eyes in our study, 40% of which was not detected with slit-lamp biomicroscopy and 63.3% of which was not evident in FA. None of the serous foveal detachments could be detected during slit-lamp biomicroscopy or FA. CONCLUSIONS: OCT-3 provided objective documentation of foveal structural changes in eyes with diabetic retinopathy. Best-corrected visual acuity provided a significant correlation with the retinal thickness at the central fovea. These results indicate that OCT can facilitate deciding on the treatment protocol (surgical or medical) and follow-up of diabetic patients, which is especially important in the early stages of diabetic maculopathy when the structural changes are not yet evident with slit lamp biomicroscopy or angiographically. PMID- 15802933 TI - Failure to detect hepatitis B virus in vitreous by polymerase chain reaction. AB - PURPOSE: To assay the vitreous of asymptomatic hepatitis B virus (HBV) carriers for the presence of HBV DNA using polymerase chain reaction (PCR). METHODS: Vitreous and serum specimens were collected from 13 carriers of HBV. The presence of HBV DNA was investigated by using PCR and Digene's Hybrid Capture System. The presence of hepatitis B surface antigen (HBsAg) in vitreous was also investigated by using the enzyme immunoassay (EIA). RESULTS: The serum was positive for HBV DNA in3 of the 13 asymptomatic carriers using PCR. Vitreous samples of all the patients, including 3 patients who were positive for HBV DNA in serum, were negative for HBV DNA with PCR and were negative for HBsAg with EIA. CONCLUSION: There is no evidence of HBV in the vitreous of asymptomatic HBV carriers. PMID- 15802934 TI - Long-term patient satisfaction of external dacryocystorhinostomy. AB - PURPOSE: To evaluate patient's opinion on the long-term success of external dacryocystorhinostomy using the same quantitative parameter pre- and postoperatively. METHODS: A postal questionnaire was sent to 139 patients who had undergone external dacryocystorhinostomy. The only parameter for success of the treatment was the improvement of patient's subjective pre- and postoperative symptoms score. The duration of this follow-up ranged from 1 year up to 5 years postoperatively. Statistical analyses were performed using the Wilcoxon Signed Ranks test and the chi2 test. RESULTS: According to the patient's own evaluation, external dacryocystorhinostomy is a successful operation. After 1 year the success percentage was 89%, after 2-3 years it was 79% and after 4-5 years it was 71%. CONCLUSION: External dacryocystorhinostomy is appreciated by the patients and considered a successful operation. The subjective evaluation of this operation should yield more creditability than objective methods. PMID- 15802935 TI - Detection of HIV RNA levels in intraocular and cerebrospinal fluids in patients with AIDS-related cryptococcosis. AB - The objective of the present study was to evaluate human immunodeficiency virus (HIV) levels in the aqueous humor and vitreous fluid in patients with and those without ocular involvement due to AIDS-related cryptococcosis. We also assessed whether cryptococcosis infection in the central nervous system (CNS) was associated with elevated HIV levels in the cerebrospinal fluid (CSF). From 1993 to 2003, we obtained 16 CSF samples from 9 AIDS patients with cryptococcal meningitis and 7 AIDS patients without CNS opportunistic infection. Samples of intraocular fluids were obtained from all 9 patients with cryptococcal meningitis. Five cases presented with ocular involvement in patients with meningitis. By using the method of reverse transcriptase-polymerase chain reaction, we detected higher HIV loads in aqueous humor (27,244+/-4,123 copies/ml) and vitreous fluid (84,930+/-5,071 copies/ml) in patients with concomitant CNS and ocular involvement due to AIDS-related cryptococcosis (p<0.05). HIV levels in the vitreous fluid were correlated with levels in CSF (r=0.77). Mean HIV level in the CSF (209,761+/-18,787 copies/ml) was significantly elevated in AIDS patients with cryptococcoal meningitis (p<0.05). To our knowledge, this is the first report to study the level of HIV loads in the CSF and intraocular fluid simultaneously in AIDS patients with cryptococcosis. Our results revealed the intrathecal and intraocular HIV replication in patients with cryptococcosis. PMID- 15802936 TI - Adrenomedullin and leptin levels in diabetic retinopathy and retinal diseases. AB - PURPOSE: Proliferative and vascular retinal diseases are important cause of irreversible blindness. Consistent features of these diseases are endothelial dysfunction and angiogenesis. Adrenomedullin (ADM) is a multifunctional vasorelaxant peptide. Leptin is a recently discovered metabolic peptide that regulates energy metabolism in human. In the present study, we aimed to investigate the possible roles of adrenomedullin and leptin in the pathophysiology of diabetic and proliferative diseases. METHODS: Ten patients with proliferative diabetic retinopathy (57.1 years, 5 female and 5 male) and 8 patients (51 years, 5 female and 3 male) with other retinal diseases including macular hole and epiretinal membrane were included in this study. All the patients had undergone pars plana vitrectomy for complications of the diseases. Vitreous samples were collected by vitreous tap during the vitrectomy. Adrenomedullin analysis was made by using reverse-phase high-performance liquid chromatography (HPLC). Leptin was measured by enzyme-linked immunosorbent assay (ELISA). Body mass index (BMI) [weight (kg)/height (m2)] was calculated for each group. The Mann-Whitney U test was used for statistics. RESULTS: The age, gender ratio and BMI were not substantially different between the two groups. The mean vitreous adrenomedullin levels (63.9+/-7.1 pmol/l) were significantly higher (p<0.05) in group I than in group II (34.25+/-3.0 pmol/l). Leptin levels in vitreous (4.54+/-1.6 ng/ml) were also significantly higher (p<0.05) in patients with diabetic retinopathy than in those without diabetes (1.83+/-0.5 ng/ml). CONCLUSION: Increased adrenomedullin and leptin levels in vitreous humor might be a possible newly associated factor in the course of vascular and proliferative retinal diseases. PMID- 15802937 TI - Subepidermal calcified nodules. AB - Subepidermal calcified nodules (SCNs) are uncommon, benign lesions usually presenting in childhood which occasionally involve the eyelids. Only a handful of cases have been reported in the ophthalmologic literature. We present 2 cases, one in a 7-year-old Hispanic boy, the other in a 13-year-old African American boy, with eyelid lesions which were clinically thought to be possible juvenile xanthogranuloma, but which on histopathologic examination showed the characteristic features of SCNs. PMID- 15802938 TI - Multiple visual hallucinations and pseudohallucinations in one individual patient: when the world is turning upside down and the television keeps falling to the ground while dwarfs are parading on the ceiling. AB - We report on a patient who experienced visual hallucinations and pseudohallucinations comprising palinopsia, peduncular hallucinosis, and oblique vision. The most probable etiologies of these phenomena are discussed with respect to clinical presentation and findings on computed tomography and magnetic resonance imaging. PMID- 15802939 TI - Influenza retinitis: association with influenza encephalitis. AB - PURPOSE: To report an unusual case of retinitis caused by influenza virus. CASE: A 30-year-old male claimed partial color blindness after recovery from influenza encephalitis. Visual acuity was 0.3 in the right eye and 0.03 in the left. An ophthalmoscopic fundus examination looked normal, but fluorescein angiography revealed granular hyperfluorescence with multiple dark circular lesions at the posterior pole of both eyes. Serological testing revealed an influenza A virus infection. With corticosteroid pulse therapy (3 days) followed by oral methyl prednisolone for 1 month, visual acuity gradually recovered. Over the following 2 year period, visual acuity has recovered to 1.2 in both eyes, but color sensation still remains impaired. CONCLUSIONS: Influenza encephalitis can cause visual loss due to retinitis at the posterior pole. PMID- 15802940 TI - Transforming growth factor-beta-1 variants are not associated with chronic nonalcoholic pancreatitis. AB - BACKGROUND: Pancreatic fibrosis is a key pathological feature of chronic pancreatitis. In vivo and in vitro data have demonstrated that pancreatic stellate cells (PSC) play a central role in pancreatic fibrosis. PSC activation and collagen synthesis are highly controlled by transforming growth factor-beta-1 (TGF-beta1). We evaluated whether functionally relevant genetic variants of TGF beta1 are associated with chronic nonalcoholic pancreatitis. METHODS: The promotor as well as exon 1 variants of the TGF-beta1 gene (G-800A, Leu10Pro and Arg25Pro) were investigated. Forty-two CP patients with a family history of nonalcoholic chronic pancreatitis (group A) and 88 patients without a family history of nonalcoholic chronic pancreatitis (group B) were studied. One hundred blood donors served as controls (group C). RESULTS: The allelic frequencies of G 800A, Leu10Pro and Arg25Pro were 12, 38 and 6% in group A; 7, 40 and 6% in group B and 12, 29 and 3% in group C, respectively. The differences were not significant. CONCLUSION: Functionally relevant genetic variants of the TGF-beta1 gene are not associated with nonalcoholic chronic pancreatitis. PMID- 15802941 TI - Pancreatic necrosis infection due to Lactobacillus paracasei in an immunocompetent patient. AB - An immunocompetent patient recovering from gallstone-induced pancreatitis had to be readmitted due to abdominal pain, fever, and rapid deterioration. Computed tomography guided needle aspiration established the diagnosis of pancreatic necrosis infection; microbiological investigations revealed monoinfection with Lactobacillus paracasei subspecies tolerans. To our knowledge, this is the first description of a pancreatic necrosis infection due to L. paracasei in an immunocompetent patient without any known risk factors for Lactobacillus infections. PMID- 15802942 TI - A temperamental model of borderline personality disorder and its implications for psychotherapy. PMID- 15802943 TI - Defining the mechanisms of borderline personality disorder. AB - Understanding the biological connections to mental processes was one of the original goals of psychoanalysis, and the development of cognitive and affective neuroscience and its methods might contribute to actualizing this goal. Personality disorders provide an opportunity to examine the complex mental structures of individuals experiencing extreme difficulties in interacting with their social environment. We provide initial information on a collaboration exploring an approach to one of the most serious personality disorders, borderline personality disorder, based upon the study of normal attention, individual differences in temperament, self definition and attachment organization, with the potential to illuminate the psychology and psychobiology of the disorder and to contribute to psychotherapeutic intervention. This developing model of borderline personality disorder can relate the symptoms to more enduring temperamental aspects of the patients. The goal is to understand the development of neural networks that underlie the abnormalities of adults, and eventually work out the interaction between temperament, genes, and experience that produce the disorder, and potentially inform intervention. PMID- 15802945 TI - The association between attentional and executive controls in the expression of borderline personality disorder features: a preliminary study. AB - BACKGROUND: Basic neurocognitive functions such as attention and executive cognitive control represent promising endophenotypes that may improve understanding of the development and expression of borderline personality disorders (BPD). We evaluated the association between performance on the Attention Network Task (ANT) and the Wisconsin Card Sorting Test (WCST), two neurocognitive laboratory instruments, and the extent of BPD psychopathology. SAMPLING AND METHODS: We studied 22 BPD-diagnosed individuals who were independently administered these two laboratory assays. Performance on these tests was used as a predictor of the extent of BPD psychopathology. RESULTS: Indexes of the ANT and the WCST were correlated with one another in this sample. Further, the extent of impairment in attention networks, specifically the orienting network, was associated with a greater spectrum of BPD psychopathology, independent of the effects of age and medication status. Finally, ANT and WCST performance were uncorrelated with general psychosocial functioning in this sample, implicating relative specificity to the extent, as contrasted with functional severity, of BPD psychopathology. CONCLUSIONS: Attentional and executive functions are promising endophenotypic markers of BPD psychopathology. The implications of these findings are considered from developmental, experimental, and clinical perspectives. PMID- 15802944 TI - Attachment and borderline personality disorder: implications for psychotherapy. AB - BACKGROUND: Psychopathology researchers and theorists have begun to understand fundamental aspects of borderline personality disorder (BPD) such as unstable and intense interpersonal relationships, feelings of emptiness, bursts of rage, chronic fears of abandonment, intolerance for aloneness, and lack of a stable sense of self as stemming from impairments in the underlying attachment organization. In the present study, we will examine self-reported attachment in a study group of well-characterized patients reliably diagnosed with BPD. SAMPLING AND METHODS: Ninety-nine outpatients reliably diagnosed with BPD using the International Personality Disorders Examination, completed a number of attachment measures including the Relationship Questionnaire, Relationship Style Questionnaire, and Experiences in Close Relationships inventory. RESULTS: Factor analysis revealed six factors that clustered into three groups corresponding to an avoidant attachment pattern, a preoccupied attachment pattern, and a fearfully preoccupied pattern. The preoccupied pattern showed more concern and behavioral reaction to real or imagined abandonments, whereas the avoidant group had higher ratings of inappropriate anger. The fearfully preoccupied group had higher ratings on identity disturbance, although only at the trend level. CONCLUSIONS: The psychometric properties and response characteristics of the ECR items suggest that the scales, keying, and domains are appropriate for assessment of attachment in BPD samples. The scales generally retain their factor structure and show a similar pattern of correlations and inter-relationships. Nevertheless, consistent with a developmental psychopathology model, there are some important differences in factor structure, indicating the need to look at both typical and atypical samples when constructing models of attachment. Further research is needed to delineate the prognostic and prescriptive significance of attachment patterns for treating patients with BPD. PMID- 15802946 TI - The construct of effortful control: an approach to borderline personality disorder heterogeneity. AB - BACKGROUND: The present study investigated the heterogeneity of DSM-IV borderline personality disorder (BPD) diagnosis as a function of the construct of effortful control. We hypothesized 3 subgroups of BPD patients based on effortful control, that would also differ in other areas of functioning, such as symptoms, interpersonal relations and personality organization. SAMPLING AND METHODS: Forty seven clinically referred individuals were reliably diagnosed as meeting DSM-IV criteria for BPD using semistructured interviews. Effortful control, symptomatology, interpersonal functioning and personality organization were assessed using self-report questionnaires. RESULTS: Cluster and profile analyses were performed and identified 3 subgroups. Subgroup 1, with high effortful control, exhibited the fewest problems in symptoms, interpersonal functioning and personality organization. Subgroup 3, with low ratings of effortful control, had the most problems in these areas, and subgroup 2, a group high in some aspects of effortful control but low in others, ranged midway between groups 1 and 3. DISCUSSION: The findings indicate a relationship between attentional mechanisms and the clinical expression of borderline personality pathology. Effortful control is a valuable construct for identifying subgroups of BPD patients, thus helping to understand the heterogeneity in BPD. Limitations of the study include the exclusive use on self-report of effortful control, as well as the small sample size. Future research should further investigate the associations of neurocognition and borderline pathology, as well as different approaches to treatment of the different BPD subgroups. PMID- 15802947 TI - Editorial: 2005 starts with a bounty of reviews and articles. PMID- 15802948 TI - Guest editorial: update on enteric viruses. PMID- 15802949 TI - Pathogenesis of astrovirus infection. AB - Astroviruses are one of the leading causes of diarrhea worldwide. In spite of its impact on human health, little is known about astrovirus pathogenesis. One reason for this may be the lack of a suitable small animal model for infection. In recent years, there has been increasing information on the mechanism of astrovirus-induced disease in mammals (including humans) and birds. This review summarizes our current state of knowledge on astrovirus pathogenesis. PMID- 15802950 TI - Immunity and resistance to astrovirus infection. AB - Astroviruses are one of the leading causes of acute viral enteritis in infants, and are recognized as a clinically important pathogen in the elderly and the immunocompromised. In spite of this, we still know very little about the immune response to astrovirus infection. Clinical observations and human volunteer studies have indicated a role for the humoral response and suggest neutralizing antibodies are important in limiting infection. Studies of human intestinal biopsies have suggested that cellular immunity; specifically CD4(+) T-cells may also be involved in the anti-astrovirus response. Additionally, various animal models have indicated potential roles for the innate immune system in controlling infections. How these various effector arms of the immune system collaborate to result in immunity and resistance to astrovirus infection is still unknown. This review summarizes our current understanding of the immune response to this pathogen and highlights the key concepts that still need to be addressed. Until we understand the role of the immune system in astrovirus infection or other enteric viruses, we will continue to be limited in our ability to treat and control gastrointestinal diseases. PMID- 15802951 TI - Identification of structural domains involved in astrovirus capsid biology. AB - Coat proteins of non-enveloped, icosahedral viruses must perform a variety of functions during their life cycle such as assembly of the coat protein subunits into a closed shell, specific encapsidation of the viral nucleic acid, maturation of the capsid, interaction with host receptors, and disassembly to deliver the genetic information into the newly infected cell. A thorough understanding of the multiple capsid properties at the molecular level is required in order to identify potential targets for antiviral therapy and the prevention of viral disease. The system we have chosen for study is the astrovirus, a family of icosahedral, single-stranded RNA viruses that cause disease in mammals and birds. Very little is known about what regions of the coat protein contribute to the diverse capsid functions. This review will present novel structural predictions for the coat protein sequence of different astrovirus family members. Based on these predictions, we hypothesize that the assembly and RNA packaging functions of the astrovirus coat protein constitutes an individual domain distinct from the determinants required for receptor binding and internalization. Information derived from these structural predictions will serve as an important tool in designing experiments to understand astrovirus biology. PMID- 15802952 TI - Rotavirus NSP4: a multifunctional viral enterotoxin. PMID- 15802954 TI - Induction and inhibition of innate inflammatory responses by adenovirus early region proteins. AB - First-generation adenovirus (Ad) gene therapy vectors deleted for the E1A, E1B, and E3 regions and carrying foreign genes under the control of strong foreign promoters induce high-level innate inflammatory responses within the first 24 hrs after transduction. Both uptake of the capsid and expression of gene products encoded by the vector contribute to the innate inflammatory response. Natural infections by Ad are frequently asymptomatic, suggesting that Ad has potent methods of inhibiting inflammation. The inability of Ad vectors to counter inflammatory responses suggests that the products of the Ad genes deleted in vector construction play critical roles in inhibiting these responses. Genetic analysis of the roles of Ad early region gene functions in vivo demonstrated that a virus made replication-incompetent by deletion of the preterminal protein gene and deleted for the transcriptional activation function of E1A effectively inhibits the innate inflammatory processes induced by Ad vectors. The mechanism(s) by which the Ad early region proteins inhibit inflammation is complex, as certain early region proteins can promote as well as inhibit inflammation, depending on the genetic context of the virus. Understanding of the roles of the Ad gene products in the induction and inhibition of innate inflammatory functions offers potential for the development of non-inflammatory vectors as well as for understanding of the mechanisms by which inflammation is regulated. PMID- 15802956 TI - Porcine reproductive and respiratory syndrome virus infection increases CD14 expression and lipopolysaccharide-binding protein in the lungs of pigs. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) is a respiratory virus of swine that plays an important role in multifactorial respiratory disease. European strains of PRRSV cause mild or no respiratory signs on their own, but can sensitize the lungs for the production of proinflammatory cytokines and respiratory signs upon exposure to bacterial lipopolysaccharides (LPS). The inflammatory effect of LPS depends on the binding to the LPS receptor complex. Therefore, we quantified the levels of CD14 expression and LPS-binding protein (LBP) in the lungs of pigs throughout a PRRSV infection. Twenty-four gnotobiotic pigs were inoculated intranasally with PRRSV (10(6) 50% tissue culture infectious doses per pig, Lelystad strain) or phosphate-buffered saline (PBS), and euthanized 1-52 days later. Lungs were examined for CD14 expression (immunofluorescence and image analysis), LBP (ELISA), and virus replication. PRRSV infection caused a clear increase of CD14 expression from 3 to 40 days post inoculation (DPI) and LBP from 7 to 14 DPI. Both parameters peaked at 9-10 DPI (40 and 14 times higher than PBS control pigs, respectively) and were correlated tightly with virus replication in the lungs. Double immunofluorescence labelings demonstrated that resident macrophages expressed little CD14 and that the increase of CD14 expression in the PRRSV-infected lungs was probably due to infiltration of highly CD14-positive monocytes in the interstitium. As both CD14 and LBP potentiate the inflammatory effects of LPS, their increase in the lungs could explain why PRRSV sensitizes the lungs for the production of proinflammatory cytokines and respiratory signs upon exposure to LPS. PMID- 15802955 TI - Mechanisms of reovirus-induced cell death and tissue injury: role of apoptosis and virus-induced perturbation of host-cell signaling and transcription factor activation. AB - Reoviruses have provided insight into the roles played by specific viral genes and the proteins they encode in virus-induced cell death and tissue injury. Apoptosis is a major mechanism of cell death induced by reoviruses. Reovirus induced apoptosis involves both death-receptor and mitochondrial cell death pathways. Reovirus infection is associated with selective activation of mitogen activated protein kinase (MAPK) cascades including JNK/SAPK. Infection also perturbs transcription factor signaling resulting in the activation of c-Jun and initial activation followed by strain-specific inhibition of NF-kappaB. Infection results in changes in the expression of genes encoding proteins involved in cell cycle regulation, apoptosis, and DNA damage and repair processes. Apoptosis is a major mechanism of reovirus-induced injury to key target organs including the CNS and heart. Inhibition of apoptosis through the use of caspase or calpain inhibitors, minocycline, or in caspase 3(-/-) mice all reduce virus-associated tissue injury and enhance survival of infected animals. Reoviruses induce apoptotic cell death (oncolysis) in a wide variety of cancer cells and tumors. The capacity of reoviruses to grow efficiently in transformed cells is enhanced by the presence of an activated Ras signaling pathway likely through mechanisms involving inhibition of antiviral PKR signaling and activation of Ras/RalGEF/p38 pathways. The potential of reovirus-induced oncolysis in therapy of human cancers is currently being investigated in phase I/II clinical trials. PMID- 15802957 TI - Infectious bursal disease virus-induced immunosuppression in the chick is associated with the presence of undifferentiated follicles in the recovering bursa. AB - Infectious bursal disease virus (IBDV) causes an acute cytolytic infection in chicken B lymphocytes resulting in destruction of the B-cell population. Most severe depletion occurs in the bursa of Fabricius, where the immunoglobulin repertoire is developed by gene conversion. Chicks surviving IBDV infection are immunosuppressed despite repopulation of the bursa with B cells. Here we show that infection of neonatal chicks with a classical virulent IBDV strain (F52/70) causes severe bursal Bcell depletion with recovery after about one week. Two distinct types of bursal follicles developed: large reconstituted follicles and small poorly developed follicles lacking a discernible cortex and medulla. The presence of large numbers of undifferentiated follicles was associated with inability to mount antibody responses to IBDV itself and after immunization with Salmonella Enteritidis bacterin, indicating that B cells in these follicles are unable to produce peripheral B-cells with an effective immunoglobulin repertoire. Additionally a number of inflammatory foci were observed in the recovering bursa. These foci contained few B cells at the margins, but large numbers of CD4(+) and CD8(+) cells, scattered gammadelta(+) T-cells and macrophages, and small central aggregates of dendriticlike cells expressing the CD40 antigen. PMID- 15802958 TI - Polyclonal hypergammaglobulinemia and formation of hydrophobic immune complexes in porcine reproductive and respiratory syndrome virus-infected and uninfected pigs. AB - Infection of young conventional, domestic pigs with porcine reproductive and respiratory syndrome virus (PRRSV) strains VR2332 and JA142 resulted in a rapid, progressive increase in serum IgG reaching maximum levels of 20-30 mg/mL at about 3 weeks post infection (p.i.), which were maintained until at least 63 days p.i., whereas the level of serum IgG remained at 4-6 mg/mL in sham infected pigs. In most of the VR2332 and JA142-infected pigs hypergammaglobulimenia was associated with the formation of hydrophobic, 150-300-kDa IgG-containing immune complexes that bound in the presence of 0.1% Tween 20 to ELISA plates that were not coated with any antigen. The ELISA plate-binding activity remained low in most infected pigs, but reached high levels in some JA142-infected pigs. Binding of the immune complexes was also observed, but at a lower level, to uncoated ELISA plates in the peptide ELISA for anti-PRRSV Abs. The immune complexes bound to uncoated ELISA plates with a much lower affinity than Abs to plates coated with peptides containing the appropriate epitopes. The immune complexes also bound to HerdChek ELISA plates, but because of low binding affinity for these plates, the bound complexes were removed by the repeated washes with Tween 20 solution. Overall the PRRSV-induced hypergammaglobulinemia and generation of ELISA plate-binding immune complexes resembled those observed in mice infected with the closely related lactate dehydrogenase-elevating virus (LDV) and thus, like the latter, seem a result of a polyclonal activation of B cells. We also found that sera of a group of older sows possessed high levels of IgG as well as of ELISA plate-binding immune complexes, in spite of being PRRSV infection negative by all criteria presently available. On the other hand, sera from wild hogs contained no ELISA plate-binding IgG in spite of possessing high total serum IgG levels. PMID- 15802959 TI - Pro-inflammatory responses in chicken spleen and brain tissues after infection with very virulent plus Marek's disease virus. AB - In chickens infected with virulent (v) or very virulent (vv) Marek's disease (MD) virus (MDV) strains, small to moderate increases in plasma nitric oxide (NO) levels are seen, respectively, whereas very virulent plus (vv+) strains induce very high levels in vivo. The data presented in this report show that chickens presenting with clinical neurological disease following infection with the vv+ RK 1 strain have significantly higher in vivo NO levels compared to RK-1-infected non-symptomatic chickens. Using quantitative real-time PCR (qPCR) assays, DNA was used to measure MDV copy numbers in the spleen and brain of P2a (MD-susceptible) and N2a (MD-resistant) chickens following infection with the JM-16 (v) or RK-1 (vv+) strains. RNA was used to measure inducible NO synthase (iNOS), interferon gamma (IFN-gamma), interleukin (IL)-1beta, IL-6, and IL-8 mRNA levels, in addition to MDV-specific mRNA expression using quantitative RT-PCR (qRT-PCR) assays. Viral DNA loads were found to be considerably higher in RK-1-infected chickens than JM-16-infected chickens at most time points in both organs, with viral copy numbers being two to four logs lower in the brain. Large increases in iNOS, IFN-alpha, IL-1beta, IL-6, and IL-8 were seen in the brains of RK-1 infected chickens. These data strongly support the hypothesis that pro inflammatory responses, including high levels of iNOS/NO, IFN-alpha, and pro inflammatory cytokine expression in the chicken brain, may play a major role in the neurological diseases associated with vv+MDV strains. PMID- 15802960 TI - Maintenance of CD8+ T cells during acute viral infection of the central nervous system requires CD4+ T cells but not interleukin-2. AB - The JHM strain of mouse hepatitis virus (JHMV) is rapidly cleared from the central nervous system (CNS) by CD8(+) T cells. In the absence of CD4(+) T cells, fewer CD8(+) T cells are found within the CNS in association with a coordinate increase in apoptotic lymphocytes. Previous data suggested that CD4(+) T cells may support CD8(+) T cells through secretion of interleukin-2 (IL-2). To determine the in vivo role of IL-2 during CNS infection, IL-2 signaling was inhibited via administration of a neutralizing IL-2-specific monoclonal antibody (mAb). In contrast to depletion of CD4(+) T cells, inhibition of IL-2 signaling did not influence CD8(+) T cell infiltration, effector cell function or survival within the CNS. These data suggest that the cellular immune response to acute neurotropic JHMV infection requires a distinct CD4(+) T cell component, but is independent of a requirement for IL-2 for induction, activation, recruitment, and/or maintenance of CD8(+) T cells within the CNS during acute infection. PMID- 15802961 TI - Requirements for the upregulation of interleukin-6 by herpes simplex virus infected gingival fibroblasts. AB - Interleukin (IL)-6 is an important proinflammatory and immunoregulatory cytokine expressed by various cells. This study examined the production of IL-6 by human gingival keratinocytes and gingival fibroblasts following herpes simplex virus (HSV) infection. Virus-cell interactions responsible for IL-6 induction by HSV-1 were determined. The amounts of IL-6 secreted by primary human gingival keratinocytes and gingival fibroblasts were determined using enzyme-linked immunosorbent assay. IL-6 expression in gingival fibroblasts was also determined using immunofluorescence staining. To further delineate the viral requirements for this induction, gingival fibroblasts were treated with antibody-neutralized viruses, UV- or heat-inactivated viruses or viral glycoprotein D of HSV-1 (gD-1). The results showed that infection of gingival fibroblasts, but not gingival keratinocytes, with HSV-1 induced production of IL-6. This modulation was blocked by neutralizing antibodies against HSV-1, suggesting that HSV-1 is required for this induction. Moreover, this induction was not abrogated when virus infectivity was destroyed by UV irradiation or heat, indicating that a complete viral life cycle is not required. Further studies showed that gD-1 alone was able to induce IL-6 secretion in gingival fibroblasts. Collectively, our data suggest that HSV-1 infection of gingival fibroblasts up-regulates production of IL-6 through a mechanism involving the interaction of gD-1 with cellular receptors. PMID- 15802953 TI - Immune modulation of the hypothalamic-pituitary-adrenal (HPA) axis during viral infection. AB - Compelling data has been amassed indicating that soluble factors, or cytokines, emanating from the immune system can have profound effects on the neuroendocrine system, in particular the hypothalamic- pituitary-adrenal (HPA) axis. HPA activation by cytokines (via the release of glucocorticoids), in turn, has been found to play a critical role in restraining and shaping immune responses. Thus, cytokine-HPA interactions represent a fundamental consideration regarding the maintenance of homeostasis and the development of disease during viral infection. Although reviews exist that focus on the bi-directional communication between the immune system and the HPA axis during viral infection (188,235), others have focused on the immunomodulatory effects of glucocorticoids during viral infection (14,225). This review, however, concentrates on the other side of the bi directional loop of neuroendocrine-immune interactions, namely, the characterization of HPA axis activity during viral infection and the mechanisms employed by cytokines to stimulate glucocorticoid release. PMID- 15802962 TI - Evolution of the intrahepatic T cell repertoire during chronic hepatitis C virus infection. AB - Chronic hepatitis C virus (HCV) infection is characterized by extensive infiltration of inflammatory cells in the liver, where there is a compartmentalization of HCV-reactive T lymphocytes. Previous studies have demonstrated a broad intrahepatic TCR repertoire; however, there is little information regarding the stability of this intrahepatic T cell population. We studied the T cell repertoires in sequential liver biopsy samples from five individuals with chronic HCV infection using TCR spectratype analysis; four subjects had been treated with IFN-alpha during the interval between biopsies. Transcripts from most TCRBV families were detectable in the liver tissues, and 25 85% of these had skewed spectratype profiles indicative of T cell clonal expansions. Most of the intrahepatic T cell expansions were not evident in an analysis of peripheral blood T cells collected at the same time as the liver biopsy. Detailed analysis using TCRBJ-primed run-off reactions revealed that the intrahepatic TCR repertoires were not stable within an individual, although some TCR clonotypes were maintained for at least 45 months. PMID- 15802963 TI - HIV-1 extrachromosomal 2-LTR circular DNA is long-lived in human macrophages. AB - HIV-1 extrachromosomal 2-LTR circles (cc2LTR) are rapidly lost in dividing cell populations and, therefore, might be interpreted as representing new infection and ongoing viral replication. However, recent work demonstrated that cc2LTR persist in infected, growth-arrested T cell lines beyond their predicted half life as previously determined in dividing cell populations. In this study, the evaluation of the stability of cc2LTR was extended to include primary human macrophages, a natural, non-dividing target of HIV-1. By quantitative real-time PCR, cc2LTR were found to persist out to 21 days post-infection in macrophages infected with both integrase competent and integrase- defective, recombinant HIV 1, whereas in activated CD4(+) T lymphocytes, they rapidly decreased over time. This persistence was associated with persistent, low level expression of the indicator gene, luciferase. These data suggest that the presence of HIV-1 cc2LTR in the PBMC of HIV-1-infected patients on suppressive HAART could be due either to ongoing generation of newly infected dividing cells, or persistence of circles in non-dividing cell populations where they appear to be stable. Furthermore, exrachromosomal circular DNA in this cell population could be a source of persisent viral protein expression. PMID- 15802964 TI - Modifications of T-lymphocyte subsets before and during interferon and ribavirin treatment for chronic hepatitis C infection. AB - Our purpose was to determine in HCV-infected patients whether T-lymphocyte sub populations were modified before and during interferon-alpha and ribavirin treatment, and whether this correlated with virological response. Twenty-two naive patients were given IFN-alpha 3 Million Units three times per week for 24 or 48 weeks and ribavirin. Sustained virological response corresponded to undetectable serum HCV RNA at treatment completion and 6 months later. Total blood lymphocyte counts and CD3(+)CD4(+), CD3(+)CD8(+), CD3(+)CD4(+)HLA-DR(+), and CD3(+)CD8(+)HLA-DR(+) lymphocyte subsets evaluated before, during, and after treatment were compared to values from 37 healthy subjects. At inclusion, patients and controls had similar total lymphocyte counts. CD3(+)CD4(+) counts and percentages were significantly higher in HCV patients. HLA-DR expression was also increased in CD4(+) (p < 0.0001) and CD8(+) T-cells (p = 0.0008) as compared with controls. During treatment, all lymphocyte subset counts and percentage decreased except the CD3(+)CD4(+) T-cell percentage which increased. Moreover, after 1 month of treatment, virological responders exhibited higher CD4(+) counts than nonresponders (p = 0.025), whereas they did not differ at inclusion or during the 2nd to 6th months of treatment. After treatment completion, all populations returned to baseline values. These results suggest that CD3(+)CD4(+) T-lymphocyte percentage increase under treatment could be related to IFN immunomodulation and associated with virological response. PMID- 15802965 TI - Needle-free jet injection of a mixture of Japanese encephalitis DNA and protein vaccines: a strategy to effectively enhance immunogenicity of the DNA vaccine in a murine model. AB - Combined immunization with gene-based and protein-based vaccines can increase vaccine effectiveness. We previously demonstrated, using a murine model for Japanese encephalitis (JE), that simultaneous immunization with a DNA vaccine (pcJEME) by the intramuscular route and a protein vaccine consisting of subviral extracellular particles (EPs) by the subcutaneous route provided a synergistic increase in immunogenicities of these vaccines. Here, we investigated a novel immunization protocol consisting of a single inoculation with a mixture of DNA and protein vaccines using a needle-free jet injector. Immunization of ddY mice with 1 microg of pcJEME mixed with 1 microg of EPs or a 1/100 dose of commercial inactivated JE vaccine (JEVAX) induced neutralizing antibody titers of 1:40 to 1:80 (90% plaque reduction) 6 weeks after immunization, whereas immunization with DNA or protein alone only induced low titers (< or =1:10). Co-immunization with pcDNA3, a CpGcontaining vector of the vaccine plasmid, increased immunogenicity of JEVAX to some extent. IgG1/IgG2a isotype profiles supported increased production of EPs in pcJEME-inoculated mice by needle-free injection and an adjuvant effect of the vector on immunogenicity of JEVAX. PMID- 15802966 TI - Augmentation of HIV-1 subtype C vaccine constructs induced immune response in mice by CpG motif 1826-ODN. AB - The greatest biomedical challenge of this century is to develop a preventive vaccine against Human Immunodeficiency Virus (HIV-1). For an HIV vaccine to be effective, it appears logical to develop new strategies that enhance the level of the immune response as well as steer it towards the desirable cellular type. In view of this, there is a need for rational inclusion of biological adjuvants into the HIV-1 vaccination strategies that could potentiate the immune responses both qualitatively and quantitatively. The adjuvant may include the immunostimulatory oligonucleotides containing CpG motifs, whose immunomodulatory characters are well established and represent the basis for an effective vaccine adjuvant. In our study, we investigated the use of an immunostimulatory oligonucleotide (or CpG motif), 1826-ODN to augment the immune response elicited by plasmid DNA vaccine constructs containing Indian subtype C HIV-1 envelope gp120 and gag protease genes in Balb/c mouse model system. A dose of 2-microg CpG motifs/mouse was found to be optimum when co-administered with the DNA vaccine constructs with the peak level of humoral and cell mediated immune responses at 6 weeks post immunization. Murine IFN-gamma ELISpot assay demonstrated that the use of 1826 ODN led to a broad based and long term recognition of the subtype C envelope and gag peptides. The use of CpG motifs has been effective in augmenting the immune responses generated by the DNA constructs. Taken together, these results are an important advancement towards the design of future preclinical and clinical trials of these vaccine constructs. PMID- 15802967 TI - Changes in Bcl-2 expression in vaccinia virus-infected human peripheral blood monocytes. AB - In this study, we show that Bcl-2, one of the most important antiapoptotic agents, is expressed in a phase-dependent manner in the human adherent monocytes after vaccinia virus infection, reflecting the viral infection stages. Early viral infection induced Bcl-2 expression in a level higher than in control cells. At 14 h post-infection (p.i.), the Bcl-2 level measured in the whole cell extracts dramatically decreased, followed by the increase at 24 h p.i. The levels of active dephosphorylated Bcl- 2 protein present in the cells reflected the gene expression character, but were much lower than in case of a heat shock. The dramatic increase of Bcl-2 protein level in the nuclear fraction at 4 h p.i. was observed. Changes in Bcl-2 mRNA content in elutriated human blood monocytes isolated from the same donor showed different kinetics, increasing up to 12 h p.i. and diminishing to undetectable level at 24 h p.i. concomitantly with a severe increase in the number of dead cells. The results indicate that virally infected adherent monocytes remain resistant to apoptosis, while freshly isolated monocytes undergo apoptotic cell death. These results throw new light on the apoptotic mechanism in the monocyte-derived cells after vaccinia virus infection in vitro. PMID- 15802968 TI - Frequency of the CCR5-Delta32 mutant allele is not increased in Belgian hepatitis C virus-infected patients. AB - A 32-base pair deletion in the CC-chemokine receptor 5 gene (CCR5), associated with resistance to human immunodeficiency virus type 1 (HIV-1) infection, has recently been suggested to act as an adverse host factor in hepatitis C virus (HCV) infection. To examine this hypothesis, we determined the CCR5-Delta32 allele frequency by polymerase chain reaction in a Belgian cohort of 163 HCV infected patients and 310 healthy control subjects. The resulting CCR5-Delta32 allele frequencies were 0.080 and 0.119 for the patient group and control group, respectively. In contrast with a previous study, we could not show a statistically significant difference between the CCR5-Delta32 allele frequencies in HCV patients and controls. Moreover, genotype distributions in both populations were in agreement with Hardy-Weinberg equilibrium. Our results do not support the hypothesis that the CCR5-Delta32 mutant allele is a risk factor for hepatitis C virus infection. PMID- 15802969 TI - Enhanced immunity and protective efficacy against SIVmac251 intrarectal challenge following ad-SIV priming by multiple mucosal routes and gp120 boosting in MPL-SE. AB - Previously, 39% of rhesus macaques primed orally, intranasally, and intratracheally with adenovirus (Ad)-simian immunodeficiency virus (SIV) recombinants and boosted with gp120 in monophosphoryl lipid A-stable emulsion (MPL-SE) remained aviremic or cleared or controlled viremia at the threshold of detection following SIV(mac251) intrarectal challenge (Study B). In contrast, no macaques primed orally and intranasally with Ad-SIV recombinants and boosted with gp120 in Quillaja Saponaria-21 exhibited undetectable viremia post-challenge (Study A). We conducted a detailed comparison of the studies to elucidate the effect of different vaccine regimens on induced immunity associated with the different challenge outcomes. Quantitative viral load comparisons were statistically analyzed. All immune responses were assessed at identical timepoints post-immunization, and cellular immunity was re-evaluated on cryopreserved cells from Study B macaques to match Study A data acquired with frozen cells. Study B exhibited greater protective efficacy, increased levels of p11C and p54m tetramer positive cells and a trend toward enhanced interferon gamma secreting cells in response to Env and Gag peptides, modestly enhanced serum neutralizing antibodies, and greater positivity in anti-gp120 rectal IgA and IgG antibodies. Study A macaques exhibited greater positivity in salivary IgA anti-gp120 antibodies. Thus, the vaccine regimen using oral-intranasal intratracheal priming and protein boosting in MPL-SE was superior, eliciting greater protective efficacy against pathogenic SIV(mac251) and enhanced SIV specific immunity, systemically and at rectal sites. The mechanism(s) by which binding antibodies, lacking neutralizing activity against the primary challenge virus, may contribute to protection requires further study. PMID- 15802970 TI - Virus-like particle (VLP) vaccine conferred complete protection against a lethal influenza virus challenge. AB - We have previously demonstrated the formation and release of influenza virus-like particles (VLPs) from the surface of Sf9 cells infected with either a quadruple baculovirus recombinant that simultaneously expresses the influenza structural proteins hemagglutinin (HA), neuraminidase (NA), matrix 1 (M1) and M2, or a combination of single recombinants that include the M1 protein. In this work, we present data on the immunogenicity and protective efficacy afforded by VLPs (formed by M1 and HA) following immunization of mice. VLP vaccine (approximately 1 microg HA) were formulated with or without IL-12 as adjuvant and administered twice, at two weeks intervals, by either intranasal instillation or intramuscular injection. All VLP-vaccinated and influenza-immunized control mice demonstrated high antibody titers to the HA protein; however, intranasal instillation of VLPs elicited antibody titers that were higher than those induced by either intramuscular inoculation of VLPs or intranasal inoculation with two sub-lethal doses of the challenge influenza virus (control group). Antibody responses were enhanced when VLP vaccine was formulated with IL12 as adjuvant. All mice were challenged with 5 LD50 of a mouse-adapted influenza A/Hong Kong/68 (H3N2) virus. Intramuscular administration of VLP vaccine formulated with or without IL-12 afforded 100% protection against a lethal influenza virus challenge. Similarly, intranasal instillation of VLP vaccine alone protected 100% of the mice, whereas VLP formulated with IL-12 protected 90% of the vaccinated mice. Not only do these results suggest a novel approach to the development of VLP vaccines for diverse influenza virus strains, but also the creation of multivalent vaccines by decoration of the surface of the VLPs with antigens from other pathogens. PMID- 15802971 TI - Surplus men, sex work, and the spread of HIV in China. AB - While 70% of HIV positive individuals live in sub-Saharan Africa, it is widely believed that the future of the epidemic depends on the magnitude of HIV spread in India and China, the world's most populous countries. China's 1.3 billion people are in the midst of significant social transformation, which will impact future sexual disease transmission. Soon approximately 8.5 million 'surplus men', unmarried and disproportionately poor and migrant, will come of age in China's cities and rural areas. Meanwhile, many millions of Chinese sex workers appear to represent a broad range of prices, places, and related HIV risk behaviors. Using demographic and behavioral data, this paper describes the combined effect of sexual practices, sex work, and a true male surplus on HIV transmission. Alongside a rapid increase in sexually transmitted disease incidence across developed parts of urban China, surplus men could become a significant new HIV risk group. The anticipated high sexual risk among many surplus men and injecting drug use use among a subgroup of surplus men may create bridging populations from high to low risk individuals. Prevention strategies that emphasize traditional measures--condom promotion, sex education, medical training--must be reinforced by strategies which acknowledge surplus men and sex workers. Reform within female sex worker mandatory re-education centers and site specific interventions at construction sites, military areas, or unemployment centers may hold promise in curbing HIV/sexually transmitted infections. From a sociological perspective, we believe that surplus men and sex workers will have a profound effect on the future of HIV spread in China and on the success or failure of future interventions. PMID- 15802972 TI - Rare mutations at codon 103 of HIV-1 reverse transcriptase can confer resistance to non-nucleoside reverse transcriptase inhibitors. AB - BACKGROUND: The K103N mutation in HIV-1 reverse transcriptase (RT) confers high level resistance to current non-nucleoside reverse transcriptase inhibitors (NNRTI). The prevalence and resistance profile of HIV-1 with other substitutions at RT codon 103 is less well documented. METHODS: K103 substitutions among over 70,000 clinical samples submitted for routine antiretroviral resistance testing at two independent centres were examined. Phenotypic resistance profiles of isolates harboring rare K103 variants in the absence of known NNRTI-associated resistance mutations were retrieved from Virco's correlative genotype/phenotype database. Genotyped samples with known treatment histories were retrieved from the British Columbia Centre for Excellence in HIV/AIDS database. Site-directed mutants containing K103 variants were constructed and phenotyped. RESULTS: K103N, R and S were observed in 29, 1.8, and 0.9% of Virco isolates and in 16, 1.5 and 0.4% of British Columbia isolates. K103T/Q/H substitutions were observed only rarely (<0.2%). The prevalence of unusual codon 103 substitutions remained stable over 5 years, except K103S, which increased over fourfold in both datasets. K103R/Q-containing clinical isolates remained phenotypically susceptible to NNRTI, whereas K103S/T/H-containing isolates showed over 10-fold decreased NNRTI susceptibility. Among patients with a known treatment history, K103S/T/H were observed primarily in individuals failing NNRTI-containing regimens. Site directed mutants confirmed decreased susceptibility to NNRTI in K103S/T/H containing recombinants. CONCLUSION: Variants at HIV RT codon 103 other than K103N are observed relatively rarely in clinical isolates, but K103 S, T and H confer decreased susceptibility to NNRTI. These data are relevant for interpretive genotype algorithms and in the design of assays specific to RT codon 103 mutations. PMID- 15802973 TI - HIV patients on antiretroviral therapy have high frequencies of CD8 T cells specific for Immediate Early protein-1 of cytomegalovirus. AB - OBJECTIVE: To assess the frequency and phenotype of cytomegalovirus (CMV) specific CD8 T cells in previously immunocompromised HIV patients with stable undetectable HIV viremia due to highly active antiretroviral therapy (HAART). METHODS: Twenty-one CMV-seropositive HIV patients with nadir CD4 T-cell counts < 50 x 10(6) cells/l, at least 4 years on HAART and 6 months of complete viral suppression (< 50 HIV RNA copies/ml) and 12 CMV-seropositive, HIV-seronegative age/sex-matched controls were studied. CD4 and CD8 T-cell responses to whole CMV and two HLA-A*02 restricted CMV peptides [NLV from pp65 and VLE from Immediate Early 1 (IE1)] were measured by interferon (IFN)gamma ELISpot. Phenotypes of peptide-specific CD8 T cells were determined by tetramer staining. RESULTS: In the ELISpot assay, HIV patients had significantly more CD8 T cells producing IFN gamma in response to VLE than controls, whereas numbers of NLV-specific and CMV specific IFN gamma spots were similar. Four HIV patients and one control had large VLE and/or NLV-specific CD8 T-cell populations despite the absence of CMV specific CD4 T cells. The majority of peptide-specific CD8 T cells from HIV patients and controls were CD28-, CD45RO+ and CD45RA-. However, a significantly higher proportion of VLE-specific CD8 T cells expressed perforin compared to NLV specific CD8 T cells in HIV patients. CONCLUSIONS: HIV patients had elevated numbers of IE1-specific, IFNgamma-producing perforin-positive CD8 T cells compared to controls. As IE1 is expressed early during CMV reactivation, these cells may be important for preventing CMV replication to pathogenic levels. In addition, CMV-specific CD4 T cells are not essential for maintenance of large populations of CMV-specific CD8 T cells in aviremic HIV patients on HAART. PMID- 15802974 TI - Divergent in vitro and in vivo correlates of HIV-specific T-cell responses during onset of HIV viraemia. AB - BACKGROUND: Cellular immune responses to HIV-1 have been examined mainly in peripheral blood mononuclear cells (PBMC). During onset of HIV replication and antigenaemia after discontinuation of highly active antiretroviral therapy (HAART), PBMC may theoretically contain HIV-specific T cells that are qualitatively and quantitatively different from specific T cells dominating in the tissues. PBMC responses throughout HIV immunotherapy trials may therefore be skewed during recurrent viraemia. OBJECTIVE: To compare cellular HIV-specific in vitro responses in PBMC during onset of HIV viraemia with corresponding in vivo responses, represented by classical delayed-type hypersensitivity tests (DTH). METHODS: HIV patients (n = 38), pre-immunized with four HIV-1 p24-like consensus peptides (Vacc-4x) during HAART, were subjected to a 14-week treatment interruption with recurrent HIV viraemia. Proliferative T-cell responses to Vacc 4x p24 peptides, HIV p24 protein, and cytomegalovirus (CMV) proteins were measured in PBMC. Corresponding Vacc-4x peptide DTH were expressed as skin infiltrate areas after 48 h. RESULTS: After 14 weeks without HAART, HIV-1 RNA increased to 72,500 copies/ml (median). The Vacc-4x p24 peptide- and HIV-1 p24 protein-induced T-cell proliferation concurrently decreased by 81 and 93% in PBMC during viraemia (medians, P < or = 0.03), whereas proliferative responses to CMV antigens were stable. In contrast, the Vacc-4x DTH areas, rather tended to increase by 36% (P = 0.08) and contained infiltrates dominated by proliferating T cells and macrophages. CONCLUSIONS: Divergent in vitro and in vivo HIV-specific cellular immune responses were found during recurrent HIV viraemia. The clinical relevance of both surrogate markers for HIV-related immune responses should be compared in future studies. PMID- 15802975 TI - Paradoxical CD4+ T-cell decline in HIV-infected patients with complete virus suppression taking tenofovir and didanosine. AB - BACKGROUND: Tenofovir (TDF) and didanosine (ddI) are both adenosine analogues with convenient posology, strong potency and a relatively high genetic barrier for resistance. The popularity of this combination, however, has been questioned due to concerns about pharmacokinetic interactions and increased risk of pancreatitis and hyperglycemia. Less information is available about other possible side effects. PATIENTS AND METHODS: HIV-infected individuals who initiated a protease inhibitor-sparing regimen between September 2002 and June 2003 at five hospitals, and had at least one subsequent visit within the next 12 months, always with complete virus suppression, were retrospectively assessed. Only drug-naive individuals and patients who simplified a prior successful antiretroviral regimen were analysed. RESULTS: Outcomes were analysed in 570 individuals according to treatment modality (98 drug-naive versus 472 simplified); the nucleoside analogue (NA) backbone (298 with TDF + ddI, 88 with ddI, 44 with TDF, and 140 with neither ddI nor TDF); and the third agent used (378 with non-nucleoside analogues versus 192 with NA). Significant CD4+ T-cell declines were seen in patients taking ddI + TDF with respect to all other NA combinations, including ddI or TDF separately. Patients exposed to high ddI doses or taking a third NA showed more pronounced CD4 declines. Plasma levels of ddI correlated with the extent of CD4+ T-cell loss. CONCLUSION: Patients receiving ddI + TDF-based combinations show CD4+ T-cell declines despite achieving complete virus suppression. This effect generally progresses with time. An imbalance in adenosine metabolites within CD4+ T lymphocytes may explain this phenomenon, which resembles the genetic purine nucleoside phosphorylase deficiency syndrome. PMID- 15802976 TI - Gender differences in clinical progression of HIV-1-infected individuals during long-term highly active antiretroviral therapy. AB - OBJECTIVE: To assess gender differences in the long-term clinical, virological and immunological outcomes during highly active antiretroviral therapy (HAART). METHODS: This longitudinal observational multicentre study followed 2460 HIV infected patients who had begun a protease inhibitor-based regimen for a median period of 43 months. Outcome measures were virological suppression (< 500 copies/ml), confirmed virological rebound after suppression, and death or new AIDS-defining illness (ADI). RESULTS: At baseline, 690 female patients (28.0%) had significantly lower age, higher prevalence of heterosexual contact and lower prevalence of intravenous drug use as risk factors for HIV infection compared with males. Furthermore, females had a lower number of AIDS-defining illnesses, higher CD4 cell counts and lower viral loads. No gender differences were reported in terms of proportion of patients achieving viral suppression or exhibiting rebound after achieving viral suppression. Female patients experienced reduced clinical progression during follow-up compared with males (P = 0.008) by Kaplan Meier analysis; however this difference was not significant in an adjusted analysis. In a multivariate model, the interaction between gender and risk factor for HIV or viral load showed that female drug users and female patients with a baseline HIV RNA viral load of 10(4)-10(5) copies/ml had a favourable clinical outcome compared with males (P = 0.035 and P = 0.015, respectively). CONCLUSION: No differences were found between genders in terms of virological and immunological outcomes during long-term HAART. Nevertheless, a lower risk of clinical progression was reported among female patients with intermediate baseline viral load than in males. PMID- 15802977 TI - Hepatic steatosis and antiretroviral drug use among adults coinfected with HIV and hepatitis C virus. AB - OBJECTIVE: To ascertain the prevalence and severity of hepatic steatosis among patients coinfected with HIV and hepatitis C virus (HCV) who had been taking antiretroviral therapy (ART); to investigate if steatosis is associated with more advanced liver disease, and to identify factors that might contribute to the process. METHODS: Steatosis was assessed among a randomly selected subset of HIV HCV-coinfected patients who had received at least 2 years of ART in a cohort study at the Johns Hopkins University HIV clinic. Liver histology was evaluated by a single pathologist. The primary outcome measure was the hepatic steatosis grade, which was classified on a 5 point scale: 0, none; 1, steatosis involving < 5% of hepatocytes; 2, 5-29%; 3, 30-60%; 4 > 60%. RESULTS: Liver histology was assessed in 112 patients, 74% of whom were taking ART at the time of biopsy. The median cumulative exposure to nucleoside reverse transcriptase inhibitors and protease inhibitors was 5.8 and 3.7 years, respectively. No steatosis was detected in 60%; grades 2-4 steatosis was recognized in 18%. In multivariate analysis, steatosis was independently associated with Caucasian race, weight > 86 kg, hyperglycemia, and stavudine use. Patients with steatosis also were more likely to have greater hepatic fibrosis (P = 0.02) and necroinflammatory activity (P = 0.005). CONCLUSIONS: Steatosis was observed in 40% of HIV-HCV-coinfected patients with extensive ART exposure and was associated with more severe HCV related liver disease. Metabolic abnormalities (excess weight and hyperglycemia) and stavudine use were modifiable risk factors for steatosis in this population. PMID- 15802978 TI - Hepatitis B and HIV: prevalence, AIDS progression, response to highly active antiretroviral therapy and increased mortality in the EuroSIDA cohort. AB - BACKGROUND: Whether hepatitis B (HBV) coinfection affects outcome in HIV-1 infected patients remains unclear. OBJECTIVE: To assess the prevalence of HBV (assessed as HBsAg) coinfection and its possible impact on progression to AIDS, all-cause deaths, liver-related deaths and response to highly active antiretroviral therapy (HAART) in the EuroSIDA cohort. METHODS: Data on 9802 patients in 72 European HIV centres were analysed. Incidence rates of AIDS, global mortality and liver-related mortality, time to 25% CD4 cell count increase and time to viral load < 400 copies/ml after starting HAART were calculated and compared between HBsAg-positive and HBsAg-negative patients. RESULTS: HBsAg was found in 498 (8.7%) patients. The incidence of new AIDS diagnosis was similar in HBsAg-positive and HBsAg-negative patients (3.3 and 3.4/100 person-years, respectively) even after adjustment for potential confounders: the incidence rate ratio (IRR) was 0.94 [95% confidence interval (CI), 0.74-1.19; P = 0.61]. The incidences of all-cause and liver-related mortalities were significantly higher in HBsAg-positive subjects (3.7 and 0.7/100 person-years, respectively) compared with HBsAg-negative subjects (2.6 and 0.2/100 person-years, respectively). The adjusted IRR values were 1.53 for global (95% CI, 1.23-1.90; P = 0.0001) and 3.58 for liver-related (95% CI, 2.09-6.16; P < 0.0001) mortality. HBsAg status did not influence viral or immunological responses among the 1679 patients starting HAART. CONCLUSIONS: The prevalence of HBV coinfection was 9% in the EuroSIDA cohort. Chronic HBV infection significantly increased liver-related mortality in HIV-1-infected patients but did not impact on progression to AIDS or on viral and immunological responses to HAART. PMID- 15802979 TI - Women in couples antenatal HIV counseling and testing are not more likely to report adverse social events. AB - BACKGROUND: Couple counseling has been promoted as a strategy to improve uptake of interventions to prevent mother-to-child HIV transmission (pMTCT) and to minimize adverse social outcomes associated with disclosure of HIV status. OBJECTIVES: We tested whether women counseled antenatally as part of a couple were more likely to accept HIV testing and nevirapine in a pMTCT program, and whether they would be less likely to experience later adverse social events than women counseled alone. METHODS: A pMTCT program that included active community education and outreach to encourage couple counseling and testing was implemented in two antenatal clinics in Lusaka, Zambia. A subset of HIV-positive women was asked to report their experience of adverse social events 6 months after delivery. Couple-counseled women were compared with individual-counseled women stratified by whether or not they had disclosed their HIV status to their partners. RESULTS: Nine percent (868) of 9409 women counseled antenatally were counseled with their husband. Couple-counseled women were more likely to accept HIV testing (96%) than women counseled alone (79%); however uptake of nevirapine was not improved. Six months after delivery, 28% of 324 HIV-positive women reported at least one adverse social event (including physical violence, verbal abuse, divorce or separation). There were no significant differences in reported adverse social events between couple- and individual-counseled women. CONCLUSIONS: Couple counseling did not increase the risk of adverse social events associated with HIV disclosure. Support services and interventions to improve social situations for people living with HIV need to be further evaluated. PMID- 15802980 TI - Sex behaviour of men who have sex with men and risk of HIV in Andhra Pradesh, India. AB - OBJECTIVE: To obtain information on sex behaviour of a large sample of men who have sex with men (MSM) in India that would assist in planning HIV prevention. METHODS: Homosexual/bisexual behaviour of 6661 MSM at 62 urban-rural locations of various sizes in the Indian state of Andhra Pradesh was assessed through detailed interview. Multivariate analyses were performed to understand the associations with unprotected penetrative sex and barriers to condom use assessed. RESULTS: The average number of different male sex partners in past 4 weeks was six. In last three sex encounters with men, totalling 19 640, anal sex occurred in 16 769, at least once by 6121 (91.9%) MSM of which 3423 [55.9%; 95% confidence interval (CI), 51.7-60.1%] did not use condom at least once. A total of 2785 (41.8%) were currently married to women and 3354 (50.4%) had had vaginal/anal sex with women in the past 3 months, of which 2818 (84%; 95% CI, 81.1-86.9%) did not use a condom. Furthermore, 1585 (25.9%; 95% CI, 22.7-29.1%) had anal sex without a condom with men and also vaginal/anal sex without a condom with women. This was prevalent across urban-rural locations and its strongest association was with currently married MSM (odds ratio 15.1; 95% CI, 12.5-18.2). The predominant reason for not using a condom with women was 'do not use with regular partner' (68.4%). CONCLUSION: This high rate of unprotected penetrative sex by MSM with both men and women suggests that HIV prevention efforts in India should include a focus on MSM as well as their wives across many urban-rural locations and not only in large cities. PMID- 15802981 TI - Interplay between hepatitis C, liver steatosis and antiretroviral therapy in HIV infected patients. PMID- 15802982 TI - Rapid viral load suppression following generic highly active antiretroviral therapy in Southern Indian HIV-infected patients. AB - We prospectively studied the initial results of 6 months of generic efavirenz based therapy on the plasma viral load in 40 patients at YRG Centre for AIDS Research and Education, a tertiary HIV referral centre in southern India. The median baseline plasma viral load was 259,000 copies/ml and at 6 months 95% of patients had plasma viral loads less than 400 copies/ml. The data support the use of generic non-nucleoside reverse transcriptase inhibitor-based regimens in resource-limited settings. PMID- 15802983 TI - Decrease of Lewis frequency in HIV-infected patients: possible competition of fucosylated antigens with HIV for binding to DC-SIGN. AB - We explored the impact of human ABO glycosyltransferase and Lewis and secretor fucosyltransferase polymorphisms in HIV infection. We found that, compared with healthy blood donors, HIV-infected patients display a significant decrease in Le(a-b+) phenotype frequencies. We showed that HIV binding on DC-SIGN-transduced Jurkat cells was inhibited by fucosyl bovine serum albumin. Our results suggest a slight protective effect of Lewis b antigen on HIV infection, possibly by the competition of Lewis antigens with HIV for binding to DC-SIGN. PMID- 15802984 TI - Clonal analyses of HIV quasispecies in patients harbouring plasma genotype with K65R mutation associated with thymidine analogue mutations or L74V substitution. AB - We analysed the quasispecies at a clonal level in patients whose plasma genotypic test harboured K65R with L74V or thymidine analogue mutations (TAM). We showed that the K65R and TAM such as M41L, D67N, T215Y/D, L210W and K219E can be borne by the same virus. We found no clone bearing both K65R and L74V substitutions. Moreover, the S68G and V75I mutations are not necessarily linked with K65R, and could thus have their own resistance effect. PMID- 15802985 TI - Can a K103N HIV strain stably overcome the wild type in the absence of non nucleoside reverse transcriptase inhibitor selective pressure? PMID- 15802986 TI - The decline of incident cases of HIV-associated neurological disorders in San Francisco, 1991-2003. PMID- 15802987 TI - Conflict and changing patterns of migration from Africa: the impact on HIV services in London, UK. PMID- 15802988 TI - Lack of interaction between atazanavir and lansoprazole. PMID- 15802989 TI - Sexually transmitted reinfection with a new hepatitis C genotype during pegylated interferon and ribavirin therapy. PMID- 15802990 TI - Making medication administration safe: report challenges nurses to lead the way. PMID- 15802991 TI - Executive summary: the State of the Science on Safe Medication Administration symposium. PMID- 15802992 TI - How we think about medication errors: a model and a charge for nurses. PMID- 15802993 TI - Errors from the consumer's perspective: tragedy motivated one woman to take action. PMID- 15802994 TI - Medication errors: why they happen, and how they can be prevented. PMID- 15802995 TI - Toward safer IV medication administration: the narrow safety margins of many IV medications make this route particularly dangerous. PMID- 15802996 TI - Medication reconciliation: transfer of medication information across settings keeping it free from error. PMID- 15802997 TI - Technology and safe medication administration. PMID- 15803001 TI - A phase II trial of sequential chemotherapy with docetaxel and methotrexate followed by gemcitabine and cisplatin for metastatic urothelial cancer. AB - Administration of noncross-resistant agents in a sequential fashion may improve outcome by targeting tumor cells with different sensitivity profiles. We evaluated the toxicity and response rate of docetaxel and methotrexate (DM) followed by gemcitabine and cisplatin (GC) in patients with metastatic or unresectable transitional cell carcinoma of the urothelium. Patients received 3 cycles of DM (40 mg/m2 methotrexate on days 1 and 8 and 100 mg/m2 docetaxel on day 8 repeated every 21 days) followed by GC (1000 mg/m2 gemcitabine on days 1 and 8 and 75 mg/m2 cisplatin on day 1 repeated every 21 days). The most common toxicities were hematologic, with grade 3/4 neutropenia and thrombocytopenia observed in 4 and 2 patients, respectively. Four partial responses were observed after DM (4 of 12, 33% response rate) and 6 responses (3 partial response, 3 complete response, 6 of 9, 67% response rate) after GC for an overall response rate of 7 of 13 (54%). Three patients who progressed on DM responded to GC and 3 responders to DM achieved further response to GC. The median overall survival was 13.6 months. Although we do not recommend this particular sequence of chemotherapy for further study, the uncompromised median survival and the ability to salvage responses with GC suggest that testing of novel agents in sequence with GC is a feasible strategy. PMID- 15803002 TI - A nonplatinum combination in metastatic transitional cell carcinoma. AB - OBJECTIVE: Cisplatin-based chemotherapy is the most effective standard treatment available for patients with metastatic bladder cancer. For those patients who are unable to receive cisplatin, other effective combination treatments are needed. The objective of this research was to determine if a combination of paclitaxel and gemcitabine would be effective in patients who were unable to receive cisplatin or have failed cisplatin treatment. METHODS: Patients with histologic proven metastatic bladder cancer who had measurable disease were eligible. Patients received chemotherapy (in an outpatient clinic) with paclitaxel at 110 mg/m2 plus 1000 mg/m2 gemcitabine on days 1 and 15 of a 28-day cycle. Patients were evaluated after 2 cycles of therapy using computed tomography and bone scan imaging. RESULTS: An objective response rate was achieved in 61% of the patients (13 of 18 subjects). Fifty-five percent of the patients who were previously treated with chemotherapy had an objective response (11 of 18 subjects). The median duration of response was 5 months and the toxicity was mild. All patients with low-risk disease had a response (8 of 18 subjects), compared with a 30% response rate in the intermediate-/high-risk group (P = 0.001, 10 of 18 subjects). CONCLUSIONS: The combination of paclitaxel and gemcitabine given in an every 2-week regimen is very well tolerated and has significant activity in previously treated patients with metastatic transitional cell carcinoma. The combination of paclitaxel and gemcitabine appears to be effective for patients with good-risk disease who are unable to take cisplatin-based chemotherapy. PMID- 15803003 TI - Long-term results of radiotherapy for early-stage testicular seminoma. AB - PURPOSE: We conducted an outcome and complications analysis of patients treated with adjuvant radiotherapy (RT) for early-stage testicular seminoma over a 35 year period at the University of Florida. METHODS AND MATERIALS: All 73 patients had the classic seminoma histology. No patient received chemotherapy as initial treatment. There were 57, 14, and 2 patients with Royal Marsden stage I, IIA, and IIB disease, respectively. All received RT to the paraaortic (PAN) and pelvic fields. One-third of the patients received prophylactic RT to mediastinal/supraclavicular fields. RESULTS: Median follow up was 15 years. Median RT dose to PAN and pelvic fields was 25 Gy. Local control and cause specific survival at 20 years were 95% and 96%, respectively. Of the 3 recurrences, 1 was salvaged with chemotherapy. Subset analysis of 41 patients revealed 1 patient who developed peptic ulcer disease and 11 patients with gastrointestinal reflux symptoms after RT. Thirty-two patients fathered a child before or after RT. Cardiovascular complications were not greater in those treated with prophylactic RT to the mediastinal region. The rate of nontesticular second malignancy for the overall group was 8% at 15 years. CONCLUSIONS: Patients treated with adjuvant low-dose RT to the paraaortic and pelvic fields for treatment of early-stage seminoma enjoy an excellent long-term cure rate with minimal risk of late RT complications. PMID- 15803004 TI - Prostate biopsy volume indices do not predict for significant Gleason upgrading. AB - Significant discordance exists between biopsy and matched prostatectomy grades. This study tests the hypothesis that surrogate tumor volume indices available from biopsies could yield an improved prediction of the underlying pathologic Gleason grade. Records of 124 patients who underwent radical prostatectomy were reviewed. Biopsies were characterized by primary and secondary Gleason grade, number of positive cores, and linear tumor length. Surgical specimens were characterized by primary and secondary Gleason grade, organ-confined disease, seminal vesicle invasion, and margins. Biochemical failure (BF) was defined by a postoperative prostate-specific antigen >0.05 ng/mL. There were 28 patients (24%) who experienced biochemical failure. On multivariate analysis, only the pathologic Gleason sum (P = 0.012) and the cumulative tumor length (P = 0.050) were independently associated with BF, and only the cumulative tumor length was associated with nonorgan-confined disease (P = 0.034). For patients with a cumulative tumor length >10 mm, 49% (18 of 37) had nonorgan-confined disease and 37% (13 of 35) had BF compared with 29% (25 of 87) and 19% (15 of 80), respectively, if they had cumulative tumor length < or =10 mm (P <0.034). Overall, an exact match was seen in 39% of biopsy Gleason grades, whereas 21% were downgraded by 1 or more points, and 41% were upgraded by 1 or more points. On univariate or multivariate analysis, none of the biopsy surrogate volume indices examined achieved significance or suggested a trend in predicting for a clinically meaningful grade change. Although indices of tumor volume from prostate needle biopsies independently predict for organ-confined disease and BF after prostatectomy, none predicted for a clinically significant upgrading or downgrading of biopsies. This suggests that the correlation that exists between such volume surrogates and outcomes after surgery reflect tumor volume effects only, independently of any possible association between tumor volume and Gleason grade. PMID- 15803006 TI - Establishing schedules for repeated doses of strontium and for concurrent chemotherapy in hormone-resistant patients with prostate cancer: measurement of blood and urine strontium levels. AB - Strontium-89 (Sr-89) alone or with concurrent chemotherapy has a role in the treatment of patients with prostate cancer (PCP). The schedules for repeated doses of Sr-89 or for concurrent chemotherapy is undetermined. The objective of this study was to measure the effective half-life (Te) of Sr-89 using a detector available in a nuclear research facility. Blood and urine samples obtained from PCP treated with Sr-89 (Metastron, Amersham, U.K.) were measured for radioactivity with a High Pure Germanium (HPGe) detector in a gamma spectrometry system (Eurisys, France). Twenty-five urine and 22 blood samples were obtained from 8 patients during a period of 160 days after Metastron injection. Sr-89 radioactivity levels in blood and urine were quite low (<8.2 x 10(-3) microCi/mL) in all patients after 21 days, whereas Sr-85 (available in 0.5% of Metastron) urine and, to a lesser extent, blood radioactivity levels were moderately high and could be detected up to 160 days. Based on Sr-85 urine levels, the calculated Sr-89 Te ranged from 9.6 to 20.7 days. Sr-89 Te can be routinely calculated in PCP based on HPGe detection of Sr-85 radioactivity levels in urine. This measurement can establish schedules for either repeated doses of Sr-89 or concurrent chemotherapy. PMID- 15803005 TI - High-dose etoposide, thiotepa, and dose-adjusted carboplatin (TVCa) with autologous hematopoietic stem cell rescue as treatment of relapsed or refractory germ cell cancer. AB - A phase I/II trial with high-dose etoposide, thiotepa, and dose-adjusted carboplatin (TVCa) with autologous hematopoietic stem cell rescue (AHSCT) as treatment for patients with relapsed or refractory germ cell cancer was investigated. The phase I portion involved a dose escalation schema for carboplatin and thiotepa while keeping the dose of etoposide constant. The intended carboplatin dose was adjusted for renal function based on the glomerular filtration rate. The phase II portion of the trial evaluated the efficacy, feasibility, and safety of tandem TVCa with AHSCT. Twenty-four patients with relapsed or refractory germ cell cancer were treated in this phase I/II trial. Nine of 24 (38%) achieved a complete response. With a median follow up of 71 months (range,1-108 months), all 9 of 24 (38%) are alive and continuously disease free. There were 2 (7%) treatment-related deaths. The median time to an absolute granulocyte count greater than 0.5 x 10/L was 11 days (range, 9-20 days) on phase I and 10 days (range, 9-13 days) on phase II therapy. The median time to a platelet count greater than 20 x 10/L was 15 days (range, 12-40 days) on phase I and 14 days (range, 13-27 days) on phase II therapy. Nonhematologic toxicity was mild to moderate. A significant correlation was seen between intended carboplatin dose and actual AUC. TVCa high-dose chemotherapy is active and well tolerated in patients with relapsed or refractory germ cell cancer. PMID- 15803007 TI - Contouring the middle and inner ear on radiotherapy planning scans. AB - The purpose of this study was to establish guidelines that help radiation oncologists contour the inner and middle ear on treatment planning scans. The radiotherapy computed tomography (CT) scans of 15 previously treated patients were reviewed for the ability to identify 3 separate auditory structures. The middle ear, the cochlea, and the vestibular apparatus were identified and contoured on each scan using anatomic landmarks. The volume and maximum axial dimension of each contour were calculated. The middle ear, cochlea, and vestibular apparatus were identified on all scans. The middle ear was defined by the tympanic membrane laterally and by the interface between air and the temporal bone in all other directions. The plane of the internal auditory canal through the temporal bone was the landmark distinguishing the vestibular apparatus from the cochlea. The mean volume of the middle ear, vestibular apparatus, and cochlea were 0.58 cm3, 0.44 cm3, and 0.14 cm3, respectively. The maximum axial dimension across the contour averaged 1.57 cm for the middle ear, 1.10 cm for the vestibular apparatus, and 0.69 cm for the cochlea. A reference atlas was constructed that shows the contour of each structure on 5 consecutive CT images. Accurate identification of the middle ear and inner ear structures on radiotherapy planning scans is possible and is necessary if critical auditory organs are to be spared during radiotherapy of targets that are located near the base of the skull. The information generated in this study will help radiation oncologists contour auditory structures accurately. PMID- 15803008 TI - Role of radiotherapy for pleomorphic adenoma. AB - The purpose of this article was to evaluate the role of radiotherapy (RT) in the treatment of pleomorphic adenoma. This is a retrospective analysis of 17 patients treated with RT alone (2 patients) or combined with surgery (15 patients). Follow up ranged from 1.8 to 27.1 years (median, 9.6 years). Local control was obtained in 8 of 10 patients (80%) with subclinical disease and 3 of 7 patients (43%) with gross disease. Overall local control was obtained in 11 of 17 patients (65%). The 5- and 10-year overall local control rates were 69% and 61%, respectively. One patient died secondary to tumor extension into the central nervous system, 1 patient had recurrence of carcinoma ex-pleomorphic adenoma and died of cancer at the primary site, 5 patients died free of disease, and 10 patients were alive at last follow up. RT is relatively effective at controlling subclinical disease in patients at high risk for developing recurrence after surgery for pleomorphic adenoma. Limited data suggest that RT may also occasionally control gross disease. PMID- 15803009 TI - A phase II study of carboplatin and paclitaxel in adenocarcinoma of unknown primary. AB - BACKGROUND: Cisplatin-based combination chemotherapy is commonly used in the treatment of carcinoma of unknown primary site. Paclitaxel has shown promising activity as a single agent in a number of malignancies. This study was conducted to evaluate the efficacy and toxicity of combination carboplatin and paclitaxel in patients with adenocarcinoma of unknown primary site (ACUP). METHODS: Twenty two patients with ACUP were enrolled in the study. Patients were treated with 200 mg/m2 paclitaxel intravenously (IV) over 3 hours followed by carboplatin IV with a targeted plasma area under the curve (AUC) of 5 mg/h/mL. RESULTS: A total of 73 treatment courses were administered with a median of 5 courses per patient. Five of the 22 registered patients had a major response for an objective response rate of 23% (90% confidence interval [CI], 0.11-0.40) by intention-to-treat analysis. The median response duration was 4.1 months (90% CI, 3.6-7.1). The median survival time was 6.5 months (90% CI, 5.5-10.1) and the 1-year survival rate was 27% (90% CI, 0.11-0.42). The major toxicity observed was neutropenia, with grade 3-4 neutropenia occurring in 3 patients (14%). There were no treatment-related deaths. CONCLUSIONS: The combination of carboplatin and paclitaxel is a tolerable and moderately active regimen in ACUP. PMID- 15803010 TI - The effects of age and comorbidity on treatment and outcomes in women with endometrial cancer. AB - BACKGROUND: Although the incidence of endometrial cancer increases with age, the effect of patient age on treatment selection and outcomes is unclear. In addition, although aging is associated with increased prevalence of comorbid conditions, the extent to which comorbidities influence endometrial cancer management is not well documented. METHODS: This population-based analysis evaluates the effect of age and comorbidity on endometrial cancer treatment and outcome in a cohort of 401 patients referred to the Vancouver Island Centre, British Columbia Cancer Agency from 1989 to 1996. Treatment and 5-year actuarial overall survival (OS) and disease-free survival (DFS) were compared by age at diagnosis (<65, 65-74, and > or =75 years) and comorbidity index (Charlson score 0-1 and > or =2). RESULTS: Median follow-up time was 7.8 years. In this cohort, 148 (37%), 152 (38%), and 101 (25%) were aged <65, 65-74, and > or =75 years, respectively. Charlson comorbidity scores > or =2 were found in 18% of patients. Distributions of disease stage, tumor characteristics, and surgical therapy were similar across age and comorbidity subgroups. Standard surgery in this cohort comprised hysterectomy without routine lymphadenectomy. In stage Ic disease, the use of postoperative RT declined with advanced age (96%, 97%, and 74% in patients aged <65, 65-74, and > or =75 years, respectively, P = 0.05) and with increased comorbidities (91% and 79% in patients with Charlson score 0-1 and > or =2, respectively, P = 0.07). Among stage Ic patients aged > or =75 years, pelvic/vaginal relapse occurred in 2 of 6 patients treated with hysterectomy alone compared with 0 of 20 patients treated with postoperative radiotherapy (P = 0.006). On multivariable Cox modeling, age at diagnosis, performance status, stage, grade, lymphovascular invasion, surgery, and radiotherapy use, but not Charlson comorbidity score, were significant predictors for overall survival. CONCLUSIONS: Although surgical therapy for endometrial cancer was not influenced by age or comorbidities, reduced use of postoperative radiotherapy in stage Ic disease was observed among women with advanced age and high comorbidity index. The associated pelvic/vaginal relapse rates were higher in elderly patients not treated with radiotherapy. Chronologic age alone should not preclude patients from consideration of optimal local therapy. PMID- 15803011 TI - A phase II study of gemcitabine in patients with relapsed or refractory low-grade non-Hodgkin lymphoma. AB - PURPOSE: The purpose of this study was to determine the tolerability, clinical response rate, and time to disease progression of gemcitabine treatment in patients with low-grade non-Hodgkin lymphoma. (NHL) PATIENTS AND METHODS: Twenty patients with low-grade NHL and progression of disease after at least 1 prior treatment regimen were prospectively enrolled. The treatment regimen consisted of 1200 mg/m2 gemcitabine intravenously administered weekly for 7 weeks followed by a 1-week rest. Subsequent treatment was given weekly for 3 weeks followed by a 1 week rest and repeated for a maximal treatment of 6 cycles until disease progression or unacceptable toxicity. RESULTS: The predominant histologic subtypes among our patients were small lymphocytic (8 of 20) and follicular (7 of 20). Grade III/IV hematologic toxicity was observed in 15 of 20 patients and dose reductions or treatment delays occurred in 19 of 20 patients. Fatigue and asthenia were treatment-limiting in many patients. There were no complete or partial responses observed and only 2 patients had stable disease after 12 weeks of treatment. The average time to progression or off-study status was 2.3 months (95% confidence interval, 1.7-2.9) with 8 patients showing progression of disease. Twelve patients were taken off the study as a result of unacceptable toxicity before observed progression of disease. No patient completed the planned course of therapy. With a median follow up of 10.2 months, 10 of 20 patients remained alive. CONCLUSION: Gemcitabine as a single agent, in this dosage and schedule, has minimal clinical activity in relapsed or refractory low-grade lymphomas and was associated with considerable toxicity. Therefore, further study of gemcitabine in this setting is not justified. PMID- 15803012 TI - A phase II study of single agent gemcitabine in relapsed or refractory follicular or small lymphocytic non-Hodgkin lymphomas: a Hoosier Oncology Group Study. AB - Gemcitabine is a pyrimidine analog that is active in patients with aggressive lymphomas and Hodgkin disease. This study assessed tumor response in patients with previously treated follicular or small lymphocytic non-Hodgkin lymphoma. This was a 2-stage phase II trial with the first stage requiring 2 of 13 responses to proceed to the second stage. Gemcitabine was given as a single agent to patients with previously treated follicular or small lymphocytic lymphomas. Gemcitabine was administered at 1250 mg/m2 over 30 minutes on days 1 and 8 of a 21-day cycle for a maximum of 6 cycles. Thirteen patients were treated with 1 to 6 cycles of chemotherapy. Two patients experienced grade 4 toxicity with neutropenia. No grade 4 nonhematologic toxicity was seen. There was 1 partial response and 8 patients (61%) had either minimal response or stable disease. Single-agent gemcitabine administered at this dose and schedule produced 1 partial remission and half the patients had stable disease. However, the study had to be stopped early because of lack of meaningful response. PMID- 15803013 TI - Radiosurgery with or without whole-brain radiotherapy for brain metastases: the patients' perspective regarding complications. AB - INTRODUCTION: When cancer spreads to the brain, patients and their families report a much reduced quality of life. Whole-brain radiation therapy (WBRT) and/or radiosurgery (RS) are mainstays of management for patients with metastatic brain cancer. Despite the ubiquity of this problem, little is known about the patients' perspective regarding management-related complications. To assess the patients' perspective regarding current therapeutic options, we retrospectively surveyed 200 consecutive patients who underwent radiosurgery with or without WBRT. METHODS AND MATERIALS: The survey consisted of 10 questions that sought information on quality of life, employment status, side effects (hair loss, fatigue, memory, concentration, mood, or employment), activity level, and satisfaction. RESULTS: We tabulated 104 responses (52%). WBRT was used in 69% (72 of 104); all patients had radiosurgery. After WBRT, the following were noted: hair loss (88%; total regrowth in 24%), excess fatigue (95%), problems with short term memory (72%), long-term memory (33%), concentration (61%), and depression (54%). Sixty-three percent of patients who had WBRT thought they had side effects after treatment compared with 34% who had radiosurgery only (P <0.001). Only 5% of RS patients reported fatigue. Seventy-six percent and 56% of respondents considered RS or WBRT, respectively, a good treatment of them (P = 0.25). Only 26 patients (27%) were employed at the time of treatment and 16 remained employed (62%). Eighteen percent believed that WBRT delayed other cancer treatments. CONCLUSIONS: Patients with brain metastases often report complications of their disease or its treatment. Side effects are most pronounced in those who received WBRT. Radiosurgery had fewer reported side effects and was believed by more patients or families to be effective. Further study of clinical outcomes and quality of life is important to guide our management of patients with these common brain tumors. PMID- 15803014 TI - Predictors of tumor control in patients treated with linac-based stereotactic radiosurgery for metastatic disease to the brain. AB - OBJECTIVE: The objective of this study was to determine predictive factors for local control (LC) of brain metastases (BM) treated with Linac-based stereotactic radiosurgery (LB-SRS). METHODS: Between January 1994 and July 2001, 80 patients (126 BM) underwent LB-SRS. All patients had follow-up imaging with computed tomography (40%) or magnetic resonance imaging (60%). Most patients had either lung (41%) or renal cell (20%) cancer. The median SRS prescription dose was 18 Gy (range, 10-21 Gy). Most patients (86%) also received whole-brain radiotherapy (WBRT). LC was defined as the absence of enlargement of the BM on follow-up scans. Actuarial LC analyses were performed by the method of Kaplan-Meier and compared with the log-rank test. Factors analyzed included histology, volume, prescription dose, maximum and minimum tumor dose, target volume ratio, number of arcs and isocenters, total degrees, and WBRT. Multivariate analysis was accomplished. RESULTS: At a median follow up of 8.8 months, 11 BM failed (8.7%). The 1-and 2-year actuarial LC rates were 88.6% and 77.2%, respectively. The most significant factors correlated with LC were prescription (P = 0.0004) and minimum tumor (P = 0.002) doses, and tumor volume (P = 0.04). On multivariate analysis, the sole factor correlated with LC was minimum tumor dose (P = 0.03). CONCLUSION: Our results confirm that LB-SRS is associated with excellent LC rates in the majority of patients treated. However, particular attention should be given to minimum target dose to ensure optimal outcome. PMID- 15803015 TI - Weekly docetaxel in combination with capecitabine in patients with metastatic gastric cancer. AB - Docetaxel (T) and capecitabine (X) are active agents against gastric cancer with synergistic antitumor effects. We conducted the current phase II study to assess the response rate and toxicity of combination TX regimen in patients with metastatic gastric cancer. Eligible patients were treated with docetaxel (36 mg/m2 intravenously) on days 1 and 8 and capecitabine (1000 mg/m2 orally twice a day) on days 1-14 of a 3-week schedule until progression occurred. From December 2001 to May 2003, 55 patients with median age of 54 years (range, 22-73 years) were enrolled; 47 patients had measurable lesions. A total of 358 courses of treatment were given, with a median of 5 (range, 1-22+) per patient. Objective responses were documented in 19 of 47 patients with measurable lesions (response rate, 40.4%; 95% confidence interval [CI], 25.9-54.9), with the median response duration of 5.6 months (range, 2.1-13.6+). At a median follow up of 15.9 months for all of 55 study patients, the median time to progression and survival were 4.5 months (95% CI, 3.4-5.6) and 12.0 months (95% CI, 7.5-16.6), respectively. Hematologic toxicities were mild to moderate, and the observed grade 3 nonhematologic toxicities, the most frequent of which was stomatitis, were generally manageable. Four patients experienced pneumonitis, but all of them responded to steroid treatment. The TX regimen was relatively well tolerated and effective against metastatic gastric cancer, with the added advantage of being an outpatient regimen. PMID- 15803016 TI - Phase II study of gemcitabine plus cisplatin in patients with metastatic breast cancer: a North Central Cancer Treatment Group Trial. AB - BACKGROUND: This 2-stage, phase II cooperative group trial examined the efficacy and toxicity of 1000 mg/m2 gemcitabine plus 25 mg/m2 cisplatin weekly for 3 weeks and repeated every 28 days for patients with previously treated metastatic breast cancer. METHODS: Eligible patients had to have measurable metastatic disease. Progression on prior treatment with at least 1 chemotherapy program for metastatic disease and 1 prior anthracycline and/or taxane-containing regimen in either the metastatic or adjuvant setting was required. Patients who had received more than 2 chemotherapy treatments were not eligible for this study. RESULTS: Fifty-eight eligible patients were entered on this 2-stage study. A 38% incidence of grade 4 thrombocytopenia observed in the first stage of accrual required lowering the chemotherapy doses to 800 mg/m2 gemcitabine plus 20 mg/m2 cisplatin weekly for the first 3 weeks of a 4-week cycle during the second stage of the study. The overall response rate was 29% (95% confidence interval [CI], 11-52%) among patients receiving the original study dose and 32% (95% CI, 18-50%) for patients receiving the lower dose. In the original- and lower-dose groups, median time to progression was 30.7 weeks (95% CI, 12.7-43.4 weeks) and 26.0 weeks (95% CI, 19.0-32.1 week), respectively. Median survival of the original- and low-dose groups was 67.9 weeks (95% CI, 42.3-90.6 weeks) and 54.1 weeks (95% CI, 41.6-73.6 weeks), respectively. Hematologic toxicities were more manageable in the lower dose group, whereas the nonhematologic toxicity profile was similar in the 2 dose groups. CONCLUSIONS: The response rate of this combination of gemcitabine and cisplatin is similar to that reported by other investigators but may not differ substantially from reports with single-agent gemcitabine in this patient population. The original dose level we used had unacceptable toxicity, which required lowering the doses of both gemcitabine and cisplatin by 20% to achieve acceptable toxicity and preserve clinical activity. PMID- 15803017 TI - The effects of ondansetron and granisetron on electrocardiography in children receiving chemotherapy for acute leukemia. AB - 5-HT3 receptor antagonists, including granisetron and ondansetron, are widely used in the prophylactic treatment of chemotherapy-induced nausea and vomiting. Although the cardiac safety of granisetron and ondansetron has been investigated in several adult studies, there is no report investigating the effects of those agents on electrocardiography (ECG) in children. The effects of intravenously infused (over 30 seconds) 0.1 mg/kg ondansetron and 40 microg/kg granisetron on ECG were assessed in 22 children receiving high-dose methotrexate therapy for acute lymphoblastic leukemia. The ECG recording was obtained at before and just after the infusion, and repeated at 1, 3, 6, and 24 hours of treatment. Granisetron administration resulted in a statistically significant decrease of mean heart rate at 1 and 3 hours, and significant prolongation of mean QT and QTc dispersions at 1 hour of infusion. In patients treated with ondansetron, no meaningful change was observed. In conclusion, intravenous granisetron but not ondansetron causes clinically asymptomatic and transient changes on ECG measurements in children receiving high-dose methotrexate therapy. PMID- 15803018 TI - Merkel cell carcinoma: two case reports focusing on the role of fluorodeoxyglucose positron emission tomography imaging in staging and surveillance. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare skin neuroendocrine carcinoma frequently occurring in the head and neck area. It is known for being a locally aggressive disease with a high incidence of regional and distant metastases. Accurate initial staging and close surveillance are critical in the management of the disease. METHODS AND RESULTS: Two cases of MCC are reported in which fluorine 18-fluorodeoxyglucose positron emission tomography (FDG PET) imaging has been beneficial in directing management and predicting clinical course. Pretreatment FDG PET scans detected metastatic disease in subcentimeter lymph nodes that were not appreciated in initial computed tomography images. Posttreatment FDG PET scans predicted response to therapy with the level of FDG uptake correlating in both areas of complete response to treatment and areas of residual diseases. Finally, FDG PET imaging also detected the progression of the disease after initial treatment. CONCLUSION: FDG PET imaging is a very sensitive modality in staging, assessment of treatment response, and surveillance of MCC. Because of the rarity of MCC, multicenter study is warranted to accumulate enough cases to determine the sensitivity and specificity of FDG PET in staging and surveillance of MCC, and the impact on the management and treatment outcome. PMID- 15803019 TI - Aggressive fibromatosis. AB - The purpose of this article is to review the pertinent literature and to define the optimal treatment of patients with aggressive fibromatosis. Data indicate the likelihood of local recurrence after surgery is high, particularly if margins are positive. Moderate-dose radiotherapy alone for gross disease or after a microscopically incomplete resection yields local control rates of approximately 75% to 80%. Treatment with pharmacologic agents results in objective response rates of approximately 40% to 50%; duration of response is variable. Thus, the optimal management for aggressive fibromatosis depends on tumor location and extent. Surgery is indicated if the lesion appears to be completely resectable with acceptable function and cosmesis. Radiotherapy is indicated for patients with unresectable tumors; those with positive margins after resection should be considered for adjuvant radiotherapy depending on the location and extent of the tumor. Pharmacologic treatment should be considered for patients with progressive disease after unsuccessful local-regional therapy. PMID- 15803020 TI - Chronic myeloid leukemia in a man with papillary carcinoma of the thyroid treated with radioactive iodine. AB - Leukemia as a second malignancy after treatment of thyroid cancer is rare. Most cases reported in the literature have occurred after cumulative doses higher than 800 mCi and it is most commonly acute leukemias. We report a case of chronic myeloid leukemia (CML) occurring in a 40-year-old man 14 years after treatment of papillary thyroid carcinoma. Our patient had the longest interval between the diagnosis of CML and administration of 131I. PMID- 15803021 TI - Serum osteoprotegerin is increased in Crohn's disease: a population-based case control study. AB - BACKGROUND: There is a potential interface between osteoporosis and the chronic inflammation of inflammatory bowel disease (IBD), and the osteoprotegerin (OPG)/receptor for activated nuclear factor-kappaB (RANK)/RANK ligand (RANKL) signaling pathway may be an important mediator, although data are limited. METHODS: We conducted a population-based case-control seroassay study to look for alterations in serum OPG and soluble RANKL (sRANKL). The study population included IBD patients who were 18 to 50 years old with Crohn's disease (CD; n = 287) or ulcerative colitis (UC; n = 166), age-matched healthy controls (n = 368), and nonaffected siblings of IBD patients (n = 146). Serum OPG and sRANKL were measured by enzyme-linked immunoassay. Sex-specific reference ranges were derived from the healthy controls. RESULTS: Analysis of variance (ANOVA) confirmed significant group differences in women for mean serum OPG (P = 0.018). CD women had higher values of OPG than UC women (P = 0.028) or healthy controls (P = 0.045), whereas the other groups were similar. OPG levels were above the reference range in 13/173 (8%) of CD women, exceeding the expected proportion (P = 0.032). In contrast, no differences in OPG were seen in men between controls, CD, or UC. Estrogen use in women (P = 0.000002) and corticosteroid use in men (P = 0.026) were associated with higher OPG levels. In multivariate analysis, CD diagnosis (P = 0.031) and estrogen use (P = 0.000002) were independently associated with higher OPG levels. No group differences were seen in mean serum sRANKL measurements. CONCLUSIONS: An OPG:sRANKL imbalance with OPG exceeding sRANKL should inhibit osteoclastogenesis and promote bone formation. CD is associated with increased fracture risk, and possibly, the paradoxically higher OPG is a counterregulatory response to factors such as inflammatory cytokines, promoting high bone turnover. Alternatively, elevated OPG in CD may reflect T cell activation. PMID- 15803022 TI - NOD2/CARD15 and TNFA, but not IL1B and IL1RN, are associated with Crohn's disease. AB - BACKGROUND: NOD2/CARD15 was described as the first susceptibility gene to Crohn's disease (CD). Polymorphisms in the TNFA gene and in the IL1 gene cluster, which are associated with an enhanced chronic inflammatory response, may also play a role in the development of CD. The aim of this study was to determine the association of polymorphisms in the CARD15, TNFA, IL1B, and IL1RN genes with risk of development of CD and with the clinicopathological profile of CD patients. METHODS: In a case-control study including 235 CD patients and 312 controls (929 controls for TNFA genotyping), the CARD15 (R702W, G908R, and 1007fs), TNFA ( 308G/A and -857C/T), IL1B (-511C/T), and IL1RN (intron 2 variable number of tandem repeats) polymorphisms were genotyped. RESULTS: We observed a significant association between CD and the CARD15 polymorphisms, with an odds ratio (OR) of 2.9 [95% confidence interval (CI), 1.9 to 4.6] for carriers of 1 variant allele and an OR of 11.8 (95% CI, 3.5 to 40.4) for carriers of 2 variant alleles. Patients with CARD15 polymorphisms had more frequently ileal or ileocolonic disease location, stricturing phenotype, abdominal surgery, and no extraintestinal manifestations. The TNFA-308A/A genotype was associated with susceptibility to CD with an OR of 3.0 (95% CI, 1.2 to 7.2). TNFA-308A/A homozygotes showed a higher frequency of erythema nodosum and arthritis, colonic disease location, and absence of abdominal surgery. No associations were found with the TNFA-857, IL1B-511, and the IL1RN VNTR polymorphisms. CONCLUSIONS: These findings suggest that CARD15 and TNFA-308 genetic polymorphisms are associated with increased risk of CD displaying distinct clinicopathological profiles. PMID- 15803023 TI - Dietary n-3 polyunsaturated fatty acids reduce disease and colonic proinflammatory cytokines in a mouse model of colitis. AB - BACKGROUND: n-3 polyunsaturated fatty acids (PUFAs) reduce the severity of chronic inflammatory bowel disease, probably by means of reduction of immune cell activation or enhancement of the epithelial barrier. Using the severe combined immunodeficient (SCID) mouse model of colitis, this study examined the effect of dietary n-3 PUFAs on development of colitis and on immunologic, epithelial, and matrix parameters in the intestines of control and colitic animals. METHODS: SCID mice were fed n-3-enriched or control diet for 3 weeks before colitis induction by transplantation of CD45RB T cells and maintained on the same diet for 4 to 8 weeks. Phenotype of infiltrating cells, epithelial ZO-1 protein, and mucosal type I collagen were assessed by immunohistology and tissue cytokines by ELISA. RESULTS: Transplanted n-3-fed animals had significantly reduced pathology scores, colonic tumor necrosis factor-alpha, interleukin-12, and interleukin-1beta compared with animals fed standard diet. Proinflammatory cytokines were reduced despite a similar level of immune cell infiltration by T cells, CD11c cells, and CD11b cells. Neutrophil infiltration was significantly reduced in n-3-fed control and colitic mice, and other myeloid populations were reduced in mice on the n-3 diet. Epithelial ZO-1 expression was increased, and myofibroblast activation significantly decreased in transplanted n-3-fed animals compared with standard diet mice. Submucosal collagen synthesis was enhanced in n-3-fed mice. CONCLUSIONS: Dietary n-3 PUFAs reduced clinical colitis and colonic immunopathology in this model of colonic inflammation by decreasing proinflammatory cytokine synthesis, reducing myeloid cell recruitment and activation, and enhancing epithelial barrier function and mucosal wound healing mechanisms. PMID- 15803024 TI - Heme oxygenase-1-generated biliverdin ameliorates experimental murine colitis. AB - BACKGROUND: Heme oxygenase-1 (HO-1) seems to have an important protective role in acute and chronic inflammation. The products of heme catalysis, biliverdin/bilirubin, carbon monoxide (CO), and iron (that induces apoferritin) mediate the beneficial effects of HO-1. Blockade of HO-1 activity results in exacerbation of experimental colitis. We tested whether HO-1 has protective effects in the development of colitis and determined that specific enzymatic products of HO-1 are responsible for these effects. METHODS: Colitis was induced by oral administration of dextran sodium sulfate (5%) to C57BL/6 mice for 7 days. HO-1 was up-regulated by cobalt-protoporphyrin (5 mg/kg, intraperitoneally). Biliverdin, exogenous CO, or the iron chelator desferrioxamine was administered to other groups. RESULTS: Cobalt-protoporphyrin treatment resulted in significant up-regulation of HO-1 protein in mucosal and submucosal cells. Induction of HO-1 was associated with significantly less loss of body weight in mice with induced colitis (-12% versus -22% in the control animals, P < 0.001). Development of diarrhea and gastrointestinal hemorrhage was substantially delayed in animals in which HO-1 was induced, and mucosal injury was significantly attenuated. Administration of CO or desferrioxamine alone had no significant effects, whereas enhanced protection with lesser evidence of bowel inflammation was observed with systemic biliverdin administration (50 micromol/kg, 3 times per day, intraperitoneally). CONCLUSIONS: We conclude that heightened HO-1 expression or administration of biliverdin ameliorates dextran sodium sulfate-induced experimental colitis. Novel therapeutic strategies based on HO-1 and/or biliverdin administration may have use in inflammatory bowel disease. PMID- 15803025 TI - Environmental factors in familial Crohn's disease in Belgium. AB - BACKGROUND: Environmental factors are believed to trigger the onset of Crohn's disease (CD) in genetically susceptible individuals. The aim of this study was to assess environmental and familial factors that might be etiologically related to CD. METHODS: Twenty-one families with 3 or more affected first-degree relatives were studied, together with 10 matched control families. There were 74 patients with CD, 84 unaffected family members, and 59 controls. Family members were interviewed together at the parental home. A 176-item questionnaire delved into first symptoms, childhood vaccinations and diseases, food items, potable water supplies, social activities, travel, pets, and home and surrounding environment. Questions were directed specifically for 2 time-frames, childhood until age 20 and a 10-year epoch before the onset of first symptoms within a family. The possible factors linked to disease were evaluated using univariate and multivariate logistic regression. RESULTS: There were significantly more smokers in the patients and their relatives than in controls. Patients had more appendicitis during adolescence, ate less oats, rye, and bran than controls, and consumed more unpasteurized cheese. Patients drank significantly less tap water and more well water than controls. Clustering of cases in time occurred in 13 of the 21 affected families. CONCLUSIONS: The less frequent consumption of oats, rye, and bran and the more frequent eating of unpasteurized cheeses imitate potential dietary influences on gastrointestinal flora. More importantly, our data suggest that the drinking of well water represents an important risk factor for CD. PMID- 15803026 TI - Clinical course of distal ulcerative colitis in relation to appendiceal orifice inflammation status. AB - BACKGROUND: Although appendiceal orifice inflammation (AOI) is frequently observed as a skip lesion of ulcerative colitis (UC), its clinical significance is not clearly understood. We aimed to evaluate whether AOI is associated with the clinical course of UC. METHODS: Ninety-four patients with newly diagnosed distal UC were prospectively enrolled at the Asan Medical Center between March 1996 and October 2002. Clinical features and colonoscopic findings during the follow-up period were analyzed in relation to initial AOI status. RESULTS: Forty eight patients were found to be initially AOI-positive and 46 to be initially AOI negative. We found no difference in the baseline demographics and clinical characteristics between these two groups. The median follow-up periods for AOI positive and AOI-negative groups were 45 and 41 months, respectively. Clinical remission was achieved in all patients of each group. The cumulative risk of relapse at 1, 3, and 5 years after remission was 31.2%, 59.8%, and 69.2%, respectively, in the AOI-positive group and 17.4%, 46.5%, and 67.2%, respectively, in the AOI-negative group (P = 0.124). The cumulative risk of proximal disease extension at 1, 3, and 5 years after diagnosis was 17.9%, 24.9%, and 44.5%, respectively, in the AOI-positive group and 9.8%, 21.5%, and 43.9%, respectively, in the AOI-negative group (P = 0.522). Proctocolectomy was performed in no patients in the AOI-positive group and in 1 patient in the AOI negative group. No mortalities were observed in either group. CONCLUSIONS: In patients with distal UC, AOI may have no prognostic implications in terms of remission, relapse, or proximal disease extension. PMID- 15803027 TI - Subclinical alveolar involvement in ulcerative colitis. AB - BACKGROUND: Although pulmonary dysfunction has been described in patients with ulcerative colitis (UC), the pathogenesis remains unclear. Our aim was to study alveolar epithelial damage using technetium-99m diethylene triamine penta acetic acid (Tc-99m DTPA) aerosol scintigraphy in patients with UC but without respiratory symptoms. METHODS: We enrolled 32 patients (18 women and 14 men; mean age, 36.4 +/- 11.6 yr) with active UC, 10 patients with inactive UC (6 women and 4 men; mean age, 43.4 +/- 11.8 yr), and 31 healthy controls (24 women and 7 men; mean age, 40 +/- 10 yr). Tc-99m DTPA aerosol scintigraphy was performed on all patients and controls. The relationship between alveolar epithelial permeability and the activity, localization, and duration of the disease was studied. RESULTS: There was a significant difference between alveolar epithelial permeability results in patients with active UC and those of the controls (P < 0.001). The same correlation was also found between the patients with inactive UC and the control group (P < 0.001). There was no correlation between Tc-99m DTPA alveolar scintigraphic test results and the stage of activity, localization, and duration of the disease. CONCLUSIONS: A latent pulmonary involvement may exist in patients with active and inactive UC. The alveolar involvement may be the earliest pulmonary damage, and a DTPA clearance test may show the early changes in pulmonary epithelial permeability that precedes clinical symptoms. Increased alveolar epithelial permeability is an extraintestinal manifestation in patients with UC and is not related to the activity of the colitis. PMID- 15803028 TI - Noninvasive diagnosis of small bowel Crohn's disease: combined use of bowel sonography and Tc-99m-HMPAO leukocyte scintigraphy. AB - BACKGROUND: Crohn's disease (CD) is frequently localized in the small bowel, with the diagnosis of disease and the assessment of its extension made by ileo colonoscopy (IC) and small bowel enteroclysis (SBE). Transabdominal bowel sonography (BS) and Tc-99m-HMPAO leukocyte scintigraphy (LS) are increasingly used for the diagnosis of CD because of their minimal invasiveness, reproducibility, and acceptable costs. METHODS: From March 2000 to July 2003, we performed IC, SBE, BS, and LS in 84 patients with either suspected or known small bowel CD. RESULTS: Small bowel CD was present in 50 patients, whereas the other 34 patients received a different diagnosis. Sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy were, respectively, 98%, 97%, 98%, 97%, and 0.97 for SBE; 92%, 97%, 98%, 88%, and 0.94 for BS; and 90%, 93%, 96%, 85%, and 0.92 for LS. In addition, the combined use of BS and LS led to overall sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy of 100%, 93%, 96%, 100%, and 0.97, respectively. BS showed a fair concordance with SBE in terms of location (k = 0.71) and a correlation with the extension of the disease (r = 0.67, P < 0.001). LS showed a concordance with SBE with regard to location in about one-half the population (k = 0.54), whereas it was less effective than SBE in defining disease extension. CONCLUSIONS: BS and LS are 2 accurate techniques for the diagnosis of small bowel CD, and their combined use can be recommended as an early diagnostic approach to patients in which the disease is suspected. SBE remains the best procedure for the definition of the location and extension of the disease. PMID- 15803029 TI - Differentiation of perianal fistulas with digital subtraction magnetic resonance fistulography. AB - BACKGROUND: Pelvic magnetic resonance imaging (MRI) is accurate in identifying perianal fistulas. The exact visualization of fistulous tracts and concomitant abscesses determine the type of treatment. To improve the detection of perianal fistulas, we studied digital subtraction MR-fistulography for tissue differentiation based on signal intensity measurements. METHODS: This study included 75 patients with the clinical diagnosis of perianal fistula. All patients were analyzed by a thin-slice, high-resolution, fast low-angle shot 3 dimensional sequence in the axial plane before and after intravenous injection of gadobenate dimeglumine, followed by image subtraction. Operator-defined regions of interest were used to calculate signal intensities of the inflamed fibrous walls of fistulas, the common femoral artery, the internal and external sphincter muscles, and the gluteus muscle. The fistulas were classified according to Parks classification. RESULTS: Based on signal intensity measurements in 75 patients with perianal fistulas, diagnosed by digital subtraction MR-fistulography, a significant differentiation between fistulous tracts and anatomic structures was possible. MRI identified 116 perianal fistulas (34 intersphincteric, 33 transsphincteric, 10 suprasphincteric, and 39 extrasphincteric) and 35 abscesses. CONCLUSIONS: Digital subtraction MR-fistulography is a new, promising, noninvasive imaging technique for the detection of perianal fistulas and abscesses. PMID- 15803030 TI - Dark lumen magnetic resonance enteroclysis in combination with MRI colonography for whole bowel assessment in patients with Crohn's disease: first clinical experience. AB - BACKGROUND: Magnetic resonance enteroclysis (MRE) is a recently introduced imaging technique that assesses the small bowel with similar sensitivity and specificity as the fluoroscopically performed conventional enteroclysis. Magnetic resonance imaging colonography (MRC) seems to be a promising technique for polyp assessment in the colon. In this feasibility study, we evaluated the combination of small bowel MRI with unprepared MRC as an integrative diagnostic approach of the whole bowel in patients with Crohn's disease. METHODS: Thirty patients with known Crohn's disease were prospectively examined. No particular colonic preparation was applied. Applying the dark lumen technique in all patients, MRE and MRC were performed within 1 session using an integrative examination protocol. T2-weighted and contrast-enhanced T1-weighted sequences were acquired. Inflammation assessment (grades 0 to 2) of the colon was compared with conventional colonoscopy in 29 patient and with surgery in 1 patient. The entire colon was graded fair to good distended in all patients. In 11 of 210 evaluated colonic segments, feces hindered an adequate intraluminal bowel assessment. Twenty-three of 30 patients had complete colonoscopy as the gold standard. In 7 patients, complete colonoscopy could not be performed because of an inflamed stenosis. RESULTS: Correct grading of colonic inflammation was performed with 55.1% sensitivity and 98.2% specificity in all segments. Considering only more extensive inflammation (grade 2), the sensitivity of MRC increased to 70.2% with a specificity of 99.2%. CONCLUSIONS: The combination of MRE and MRC could improve the diagnostic value of abdominal MRI evaluation in patients with Crohn's disease. However, MRC can not replace conventional colonoscopy in subtle inflammation assessment. PMID- 15803031 TI - Infliximab and semen quality in men with inflammatory bowel disease. AB - BACKGROUND: Infliximab is effective for induction and maintenance of remission in reproductive age men with Crohn's disease. There is no available data on the effects of infliximab on semen quality. The aim of this study was to determine whether changes in semen quality occurred in men receiving infliximab. METHODS: In this prospective study, each patient served as his own control. Patients completed general health and fertility questionnaires and were assessed for disease activity. Two semen analyses were completed before infusion with infliximab and 1 semen analysis was completed 1 week after infusion. Mean semen parameters before infusion were compared with postinfusion parameters by paired t tests. RESULTS: Ten men completed the study. Seven were on maintenance infliximab (group 1) and 3 were receiving a first dose (group 2). Seven had Crohn's disease, 2 had indeterminate colitis, and 1 had ulcerative colitis. All group 1 patients were in remission. Group 2 patients had moderate or severe disease. In comparing pre- and postinfusion semen parameters in all 10 patients, there was a significant increase in semen volume (P = 0.013) after infusion with infliximab and a trend toward decreased sperm motility (P = 0.061). Group 1 had a significant increase in semen volume after infusion (P = 0.039) and a significant decrease in normal oval forms after infusion (P = 0.038). In comparing group 1 and group 2, there was a significant difference in sperm progression. CONCLUSIONS: Infliximab therapy in men may decrease sperm motility and the number of normal oval forms. Whether these findings translate into impaired fertility is an area for further study. PMID- 15803032 TI - Health-related quality of life and psychosocial functioning of adolescents with inflammatory bowel disease. AB - BACKGROUND: This study aimed to study how inflammatory bowel disease (IBD) affects health-related quality of life (HRQoL) during adolescence and to examine how self-esteem influences HRQoL. METHODS: We compared self-esteem, anxiety, and parental reports on behavioral problems in a group of IBD patients to a healthy norm group. Forty children and 38 parents filled out questionnaires separately. Trait anxiety, self-perception, and the data on the Child Behavior Checklist were taken to compare the IBD population with healthy norms, using Student's t tests and 1-sample t tests. Effect sizes were calculated to show the clinical relevance of the differences. Multiple regression analyses were performed to assess the association between disease-related variables and self-perception with HRQoL, anxiety, and problem behavior. RESULTS: The results of this study show that adolescents with IBD, especially boys, have a significantly worse HRQoL and show more internalizing problem behavior compared with healthy peers. An important predictor of HRQoL is self-esteem. CONCLUSIONS: In conclusion, adolescents with IBD are at risk for experiencing problems with their illness. Because self-esteem is an important predictor of HRQoL, it should be taken into account in future interventions for these children. PMID- 15803033 TI - Inflammatory bowel diseases at the University of Chicago--early experiences: a personal historical account. PMID- 15803035 TI - Does daily fecal volume predict the clinical outcome of the acute flare of extensive ulcerative colitis in terms of need for acute colectomy? PMID- 15803034 TI - Desensitization treatment of an aspirin- and mesalamine-sensitive patient with Crohn's disease. PMID- 15803036 TI - Productive work and employment for persons with traumatic brain injury: what have we learned after 20 years? AB - This article describes return to work outcomes for individuals with traumatic brain injury (TBI) through a selective review of the published literature and an examination of TBI labor force participation from the Rehabilitation Services Administration 91l and the Social Security Administration Benefits Planning, Assistance and Outreach databases. Implications and recommendations to further our understanding about the different parts of the work outcome profile of individuals with TBI are offered. PMID- 15803037 TI - Development of systems of care for persons with traumatic brain injury. AB - The history of the development of comprehensive systems of care for treatment of survivors of traumatic brain injury is reviewed in relation to publications in The Journal of Head Trauma Rehabilitation (JHTR). This development has drawn from multiple other areas of clinical endeavor and has extended the range of services from an acute inpatient focus to include multisite delivery and lifetime supported living aspects. Widespread implementation of this comprehensive system approach continues to lag because of coordination and financing challenges. The history and details of these developments are comprehensively reflected in the published articles of JHTR over the past 20 years. PMID- 15803038 TI - Effects of categorization training in patients with TBI during postacute rehabilitation: preliminary findings. AB - BACKGROUND: Previous research suggests that traumatic brain injury (TBI) interferes with the ability to extract and use attributes to describe objects. This study explored the effects of a systematic Categorization Program (CP) in participants with TBI and noninjured controls. PARTICIPANTS: Ten persons with moderate to severe TBI who received comprehensive postacute rehabilitation services and 13 matched noninjured controls participated in the study. INTERVENTION: All participants received CP training for 3 to 5 hours per week for 10 to 12 weeks that consisted of 8 levels and targeted concept formation, object categorization, and decision-making abilities. MAIN OUTCOME MEASURES: The Mayo Portland Adaptability Inventory-3 (MPAI-3) and the Community Integration Questionnaire (CIQ). Two Categorization Tests (administered pretraining and posttraining) and 3 Probe Tasks (administered at specified intervals during training) assessed skills relating to categorization. RESULTS: Both groups showed significant improvement in categorization performance after the CP training on the 2 Categorization Tests related to the CP. They also were able to generalize and apply categorization and sorting skills in new situations (as measured by the Probe Tasks). Participants with TBI had improved functional outcome performance measured by the MPAI-3 and the CIQ. CONCLUSIONS: The systematic and hierarchical structure of the CP is beneficial to participants with TBI during postacute rehabilitation. This study contributes to the growing body of evidence supporting cognitive rehabilitation after moderate to severe TBI. PMID- 15803039 TI - Community outcomes following traumatic brain injury: impact of race and preinjury status. AB - OBJECTIVE: To investigate the contribution of preinjury differences and potential biases in outcome measurement in explaining outcome differences between white and African American persons with moderate and severe traumatic brain injury (TBI). DESIGN: Prospective, 2 group longitudinal study with retrospective self assessment of preinjury status. SETTING: Inpatient and outpatient TBI rehabilitation program. PARTICIPANTS: Ninety-four persons with moderate and severe TBI (55 whites and 39 African Americans) who provided data on both preinjury status and 1-year outcome. MEASURES: Community Integration Questionnaire, aggression and depression subscales of Neurobehavioral Functioning Inventory-Revised, Satisfaction With Life Scale, other questions on demographic and social status; all measures were selected by a biracial focus group. RESULTS: Whites and African Americans did not differ significantly on demographic factors except gender; nor on retrospective, self-reported preinjury status on any of the outcome measures. At 1 year post-TBI, African Americans reported significantly lower social integration subscale scores than whites. African Americans may also have lost more income than did whites. All other outcome measures were comparable between groups and showed declines in community productivity, increases in depression symptoms, and lower satisfaction with life for both whites and African Americans compared to preinjury. A higher rate of change in living situation post TBI may partially account for lower levels of social integration for African Americans. CONCLUSION: Whites and African Americans who are comparable prior to injury may experience generally similar outcomes, but differences in social and financial outcomes require further investigation. PMID- 15803040 TI - An interactive Web-based intervention for persons with TBI and their families: use and evaluation by female significant others. AB - OBJECTIVE: To assess the feasibility of providing in-home adjunctive and supportive services to persons with traumatic brain injury (TBI) and their families via a Web site. DESIGN: Nineteen families were provided with access to the Web site intervention for 6 months. Those who needed it were provided with a computer and Internet service in their homes. PARTICIPANTS: Adult women who were the significant others of adult males with moderate-to-severe TBI. MAIN OUTCOME MEASURE: Value and ease of use of the Web site. Each participant's usage of the Web site was automatically tracked including each page visited, time of day, and time spent on the page. RESULTS: Female significant others found the Web site to be valuable and easy to use, and used it throughout the 6-month period. The on line support group was the most used and valued module. CONCLUSIONS: Family caregivers will use Web-based interventions to help meet their needs for social support, information, and guidance following the return home of persons with TBI. PMID- 15803042 TI - The therapeutic alliance in the treatment of personality disorders. AB - Because personality disorders are associated with significant impairment in interpersonal relationships, special issues and problems arise in the formation of a therapeutic alliance in the treatment of patients with these disorders. In particular, patients with narcissistic, borderline, and paranoid personality traits are likely to have troubled interpersonal attitudes and behaviors that will complicate the patient's engagement with the therapist. While a strong positive therapeutic alliance is predictive of more successful treatment outcomes, strains and ruptures in the alliance may lead to premature termination of treatment. Therefore, clinicians need to consider the patient's characteristic way of relating in order to select appropriate interventions to effectively retain and involve the patient in treatment. Research has shown not only the importance of building an alliance but also that this alliance is vital in the earliest phase of treatment. The author first reviews several definitions of the therapeutic alliance with reference to how they apply to the treatment of patients with personality disorders. Issues relevant to forming a therapeutic alliance with patients with personality disorders are then discussed in terms of the three DSM-IV-TR personality disorder clusters. However, the author notes that these categories do not adequately capture the complexity of character pathology and that clinicians also need to consider which aspects of a patient's personality pathology are dominant at the moment in considering salient elements of the therapeutic alliance. In dealing with Cluster A personality disorders (schizotypal, schizoid, and paranoid personality disorders), what is most relevant for alliance building is the profound impairment in interpersonal relationships. The Cluster B "dramatic" personality disorders (antisocial, borderline, histrionic, and narcissistic) are all associated with pushing the limits. Consequently, clinicians need to exercise great care to avoid crossing inappropriate lines in a quest to build an alliance with patients with one of these disorders. Patients with Cluster C "anxious/fearful" personality disorders (avoidant, dependent, and obsessive-compulsive personality disorders) are emotionally inhibited and averse to interpersonal conflict. These patients frequently feel guilty and internalize blame for situations even when there is none, a tendency that may facilitate alliance building because the patients are willing to take some responsibility for their dilemma and may engage somewhat more readily with the therapist to sort it out, compared with patients with more severe Cluster A or B diagnoses. The author then reviews considerations relevant to treatment alliance that arise in the different treatment approaches that may be used with patients with personality disorders, including psychodynamic psychotherapy/psychoanalysis, cognitive-behavioral therapies, and psychopharmacology. The author also discusses issues, especially splitting, that arise in the alliance when patients with personality disorders are treated in inpatient psychiatric hospital settings. PMID- 15803043 TI - Boundary issues and personality disorders. AB - The author first presents an overview of the basic elements of boundary theory and clarifies the distinction between boundary crossings and boundary violations. The concepts of context dependence, power asymmetry, and fiduciary duty as they relate to boundary problems are also discussed. The intrinsic and extrinsic consequences of boundary problems are reviewed. The extrinsic consequences fall into three major categories: civil lawsuits, complaints to the board of registration, and complaints to professional societies. The author then reviews types of boundary issues that arise in relation to histrionic, dependent, antisocial, and borderline personality disorders. Countertransference issues that arise in working with patients with personality disorders are discussed, as well as cultural differences that may affect the perception of boundary problems. The article ends with a list of risk management principles and recommendations for avoiding boundary problems in the therapeutic relationship. PMID- 15803044 TI - Psychodynamic perspectives of collaborative treatment. AB - In contemporary psychiatric practice, collaborative treatment between a psychiatrist and psychotherapist is very common. Much attention has been devoted to the fiscal, ethical, and legal issues relevant to this treatment modality. However, such collaboration had been proposed long before the advent of managed care, which has accelerated the trend to separate the dynamic and the biologic. While fiscal considerations have been highlighted of late, the underlying tension between the dynamic and biologic remains and with it the complexity and potential pitfalls that directly and indirectly affect patient care. These issues are often neglected or kept in the realm of the unconscious. The key issues of transference, countertransference, and dynamic issues related to medication are discussed from the perspectives of the psychiatrist, the therapist, and the patient. In this article, the authors review the literature in order to revisit and reframe psychodynamic issues, as a timely reminder for clinicians to look at patients in different frameworks, and as a means of enhancing the effectiveness of collaborative treatment. PMID- 15803045 TI - Diagnosis and assessment of sleep and circadian rhythm disorders. AB - Sleep and wakefulness are fundamental behavioral and neurobiological states that characterize all higher animals, including human beings. This article presents an overview of the current state of our knowledge concerning the function of sleep and sleep-wake rhythms, the neurobiology of circadian rhythms, how wakefulness and sleep are studied, and the clinical assessment and diagnosis of sleep and circadian rhythm disorders. Major theories of the function of sleep and sleep wake rhythms are reviewed, including ecological or environmental advantage, physical restoration, optimizing waking function, learning and integration of experience, and survival. The author then reviews what is known about the neurobiology of endogenous circadian rhythms and how they are affected by environmental time cues. How sleep is studied using polysomnography (PSG) is explained, and the PSG characteristics of the three major neurobehavioral states, wakefulness, rapid eye movement (REM) sleep, and non-rapid eye movement (NREM) sleep, are described. Systems of classifying sleep disorders are reviewed, including those of the DSM-IV-TR, the International Classification of Sleep Disorders, and the ICD-10. Methods of assessing sleep complaints are then described, including taking an accurate history from the patient and bed partner, use of sleep history questionnaires and sleep-wake diaries, use of actigraphy, and use of PSG. PMID- 15803046 TI - Drug-drug interactions: proof of relevance (part I). PMID- 15803047 TI - Flying high: the myth of specificity. PMID- 15803048 TI - Delusional disorder and the law. PMID- 15803049 TI - Personality comorbidity in adolescent females with ADHD. AB - OBJECTIVE: Attention-deficit/hyperactivity disorder (ADHD), a common disorder in adolescents, is associated with extensive comorbid Axis I psychopathology. However, few studies have addressed Axis I comorbidity in girls with ADHD, and even fewer have examined comorbid personality disorders in this population. This pilot study explored personality patterns in psychiatrically hospitalized adolescent females with ADHD. METHODS: Thirty-seven adolescent females were assessed for ADHD using the Diagnostic Interview for Children and Adolescents and assigned to groups based on the presence or absence of ADHD. The two groups (ADHD Group, n=10; No ADHD Group, n=27) were compared using the Structured Interview for DSM-III-R Personality Disorders to assess for coexisting Axis II disorders. Multiple information sources and clinical corroboration were used to arrive at "best estimate" diagnoses. RESULTS: Subjects in the ADHD Group were found to have significantly more personality disorders than those in the No ADHD Group (4.5 versus 1.59 diagnoses/subject). Paranoid, histrionic, borderline, passive aggressive, and dependent personality disorders were significantly more frequent in the ADHD group. CONCLUSIONS: In this study, adolescent girls with ADHD were more likely to have personality disorders than those without ADHD. Since extensive personality comorbidity may prolong and complicate treatment, early and complete identification of these disorders will foster effective treatment planning. PMID- 15803050 TI - Zymosan-induced generalized inflammation: experimental studies into mechanisms leading to multiple organ dysfunction syndrome. AB - Patients suffering from multiple organ dysfunction syndrome (MODS) comprise a heterogeneous population, which complicates research in its pathogenesis. Elucidation of the mechanisms involved in the development of MODS will ultimately necessitate the collection of tissue samples and the performance of invasive procedures. These requirements greatly reduce the possibilities for research in human subjects. Therefore, an animal model for MODS is a necessary and valuable tool. In the mid 1980s, the zymosan-induced generalized inflammation (ZIGI) model was introduced. Intraperitoneal injection of zymosan in mice or rats leads, in the course of 1 to 2 weeks, to increasing organ damage and dysfunction. The ZIGI model has been recognized as the one that best resembles human MODS and it has been used widely to study systemic inflammation in relation to organ failure. This review describes the ZIGI model and gives an overview of the results obtained. PMID- 15803052 TI - Terlipressin as rescue therapy for intractable hypotension due to septic shock in children. AB - Intractable hypotension due to septic shock is associated with high mortality rates in critically ill children worldwide. The use of terlipressin (triglycyl lysine-vasopressin), an analog of vasopressin with a longer duration of action, recently emerged as a treatment of hypotension not responsive to vasopressors and inotropes. This was a retrospective study set in an 18-bed pediatric critical care department in a tertiary care children's hospital. We reviewed the files of all children with septic shock who were treated with terlipressin between January 2003 and February 2004. Fourteen children (mean age, 5.6 years; range, 4 days to 17.7 years) were treated with terlipressin in 16 septic shock episodes. Significant improvements in respiratory and hemodynamic indices were noted shortly after treatment. Mean arterial blood pressure increased significantly from 54 +/- 3 to 72 +/- 5 mmHg 10 min after terlipressin administration (P = 0.001). Heart rate decreased from 153.0 +/- 6.5 beats/min to 138.0 +/- 7.5 beats/min 12 h after treatment onset (P = 0.003). Epinephrine infusion was decreased or stopped in eight patients after terlipressin administration. Urine output increased from 1.6 +/- 0.5 mL/kg/h to 4.3 +/- 1.2 mL/kg/h 1 h after treatment onset (P = 0.011). PaO2 increased from 95.1 +/- 12.3 mmHg to 110.1 +/- 20.5 mmHg, and the oxygenation index decreased from 10.2 +/- 2.2 to 9.2 +/- 1.7. Terlipressin treatment of hypotension due to septic shock was successful in eight out of 16 episodes. Six of the 14 patients with poor prognosis for survival recovered. We conclude that terlipressin improves hemodynamic indices and renal function in critically ill children. Terlipressin should be considered as a rescue therapy in intractable shock not responsive to catecholamines in children. PMID- 15803051 TI - Effects of high-dose of intravenous immunoglobulin and antibiotics on survival for severe sepsis undergoing surgery. AB - The objective of this study was to assess the impact on outcome of adjuvant therapy (high-dose of immunoglobulin [Ig] M-enriched intravenous Ig, IVIG) in intensive care unit (ICU) patients who underwent surgery by abdominal sepsis. This was a prospective, randomized, double-blind, controlled study set in the medical/surgical ICUs of seven teaching hospitals. Patients with severe sepsis and septic shock of intra-abdominal origin admitted to the ICU within 24 h after the onset of symptoms were included in the study. Polyvalent IgM-enriched Ig (Pentaglobin; IVIG group) at a dosage of 7 mL/kg/day for 5 days or an equal amount of 5% human albumin (control group) was randomized. Fifty-six patients were enrolled. The overall mortality rate was 37.5.%. Twenty patients had shock and 36 had severe sepsis (the mortality rate was 55.0% and 25.0%, respectively). In the intent-to-treat analysis, the mortality rate was reduced from 48.1% in patients treated with antibiotic (ATB) plus albumin to 27.5% (P = 0.06) for patients with ATB plus IVIG. The organ failure score (1.0 +/- 0.6 vs. 1.2 +/- 0.9), organ dysfunction score (1.7 +/- 1.1 vs. 1.8 +/- 1.0), and reoperation rate (17.2% vs. 29.6%) were not different between IVIG and control groups, respectively. Eight patients (14.3%) received inappropriate ATB initial therapy (IAT), and seven died (87.5%). IAT was the only variable independently associated with death (odds ratio, 19.4) in a logistic regression model. We conclude that IVIG administration, when used in combination with adequate antibiotics, improved the survival of surgical ICU patients with intra-abdominal sepsis. The initial choice of antibiotic has a dramatic impact on outcome. PMID- 15803053 TI - Opposing effects of tumor necrosis factor receptor 1 and 2 in sepsis due to cecal ligation and puncture. AB - Tumor necrosis factor (TNF)-alpha, a cardinal molecule in the cascade of sepsis induced host injury, binds to two distinct receptors: tumor necrosis factor receptor (TNFR) 1 and TNFR2. We used the cecal ligation and puncture model of polymicrobial sepsis to elucidate the role of these receptors in sepsis pathogenesis. Mice lacking TNFR1 had prolonged survival with less hypothermia, whereas mice lacking TNFR2-/- had shortened survival and more profound hypothermia than wild-type mice. TNFR1-/- and TNFR2-/- mice had increased serum concentrations of interleukin (IL) 1beta and total TNF-alpha (free plus receptor bound) compared with wild-type mice, but there were no differences in IL6 or IL10 concentrations. Furthermore, free TNF-alpha was markedly elevated in the serum and peritoneal fluid of mice lacking TNFR2, supporting a role for this receptor in regulating the concentration of TNF-alpha. Lastly, apoptosis of ileal crypt epithelial cells was increased in mice lacking TNFR1, but there were no differences in lymphocyte apoptosis. These data suggest that in sepsis, TNFR1 mediates much of the TNF-alpha-induced pathology, whereas TNFR2 mediates protective effects. PMID- 15803054 TI - Nitric oxide from inducible nitric oxide synthase sensitizes the inflamed aorta to hypoxic damage via respiratory inhibition. AB - We tested whether nitric oxide (NO) could synergize with hypoxia to induce damage to the aorta isolated from rat. We found that 4 h of mild hypoxia (5% O2) caused substantial necrosis of isolated rat aortae (measured as lactate dehydrogenase release) if inducible NO synthase (iNOS) had previously been induced by endotoxin plus interferon-gamma. Mild hypoxia caused no significant necrosis in the absence of this inflammatory activation, and inflammatory activation caused little damage at a higher oxygen levels (21% oxygen). An iNOS inhibitor (1400W) prevented the necrosis induced by inflammation plus mild hypoxia, whereas the NO donor diethylenetriamine (DETA)/NO adduct, 0.5 mM) greatly sensitized the noninflammed aorta to necrosis induced by mild hypoxia. NO inhibited aortic respiration to a greater degree at lower oxygen concentrations, consistent with NO inhibition of cytochrome oxidase in competition with oxygen. A specific inhibitor of mitochondrial respiration, myxothiazol, caused necrosis of aortae over a similar time course to NO. DETA/NO plus mild hypoxia-induced cell death was substantially reduced by a glycolytic intermediate 3-phosphoglycerate, suggesting that necrosis resulted from energy depletion secondary to respiratory inhibition. This NO induced sensitization of aorta to mild hypoxia may be important in sepsis and other pathologies where iNOS is expressed. PMID- 15803055 TI - Mesenteric vascular dysfunction after cardiopulmonary bypass with cardiac arrest is aggravated by coexistent heart failure. AB - Although patients suffering from heart failure (HF) have an increased incidence of nonocclusive mesenteric ischemia after opened heart surgery, the impact of cardiopulmonary bypass with cardiac arrest (CPB) on mesenteric vascular circulation in such situation remains unexplored. Therefore, the present study investigates the effects of CPB on mesenteric vascular reactivity, regional metabolism, and oxidative stress in an experimental model of HF. Volume-overload HF was induced in six dogs by bilateral femoral arteriovenous fistula. Six sham operated dogs were used as controls. Eight weeks later, the short-term effects of 90 min of CPB were assessed in vivo during acute experiments. The significant increase in left ventricular end-diastolic volume in HF animals did not influence the vasodilator response of the superior mesenteric artery to acetylcholine (ACH) and nitroprusside (SNP) under baseline conditions. However, reduced mesenteric oxygen delivery, increased oxygen extraction, and lactate release were found during CPB in the HF group. In addition, an increased free radical production was assessed in the HF group during (89 +/- 23 x 10 relative light units [RLU]) and after CPB (93 +/- 15 x 10 RLU) compared with controls (45 +/- 15 and 49 +/- 7 x 10 RLU, respectively). Finally, 90 min of CPB led to a more pronounced decrease of ACH- (-22% +/- 5% vs. -42% +/- 9%, P < 0.05) and SNP- (-14% +/- 4% vs. -50% +/ 7%, P < 0.002) induced mesenteric vasodilations in the HF group compared with controls. We conclude that coexistent HF significantly enhances the pathological effects of CPB on the mesenteric vascular circulation by additionally altering endothelial and smooth muscle vascular function consequent to augmented oxidative stress. PMID- 15803056 TI - Lymphatic system as a path underlying the spread of lung and gut injury after intestinal ischemia/reperfusion in rats. AB - We investigated in rats the influence of the lymphatic system and of tumor necrosis factor (TNF) on the lung inflammation resulting from intestinal ischemia/reperfusion (I/R) performed by 45-min occlusion of the superior mesenteric artery followed by 2 h of reperfusion. A group of rats had the thoracic lymph duct ligated before I/R. In lungs, intestinal I/R evoked a significant neutrophil recruitment, and enhanced microvascular permeability, in addition to generation of TNF in serum. In the gut, there was lowered lactate dehydrogenase (LDH) activity and increased microvascular permeability. Upon lymph duct ligation, I/R rats had a significant reduction of pulmonary neutrophil recruitment and plasma extravasation, in addition to high amounts of TNF in the lymph, contrasting with undetectable levels in the serum. In addition, LDH gut levels in these animals were close to basal values; there was also some (yet significant) reduction of microvascular permeability, suggesting that the ligation of the lymphatic duct exerted some degree of protection against the intestinal injury caused by I/R. In I/R rats, the treatment with pentoxifylline (PTX) reduced TNF in serum and blunted other lung alterations. The gut alterations caused by intestinal I/R were largely blocked by PTX. On the other hand, in I/R rats with lymph duct ligation, PTX exacerbated the reduction of pulmonary neutrophil recruitment, but did not affect pulmonary and intestinal microvascular permeabilities. Similarly, intestinal LDH activity and serum TNF levels were unaffected. Overall, our data show that the pulmonary and gut injuries induced by intestinal I/R are partially dependent on TNF, which is conceivably generated in the injured gut tissue due to intestinal I/R and carried by the lymphatic system. Thus, the mesenteric lymphatic drainage seems to play a role as a path modulator of the pulmonary and intestinal dysfunctions that follow a gut trauma. PMID- 15803057 TI - Choice of fluid influences outcome in prolonged hypotensive resuscitation after hemorrhage in awake rats. AB - Hypotensive resuscitation (Hypo) has been considered an alternate resuscitation strategy in clinical settings that prevent the application of standard Advanced Trauma Life Support care. However, validation of this approach when used for prolonged periods of time remains to be demonstrated. The purpose of this study was to evaluate prolonged Hypo as an alternative to standard resuscitation using various currently available resuscitative fluids. Unanesthetized, male Sprague Dawley rats underwent computer-controlled hemorrhagic shock and resuscitation. There were six experimental groups; nonhemorrhage (NH), nonresuscitated control (C), Hypo with lactated Ringer's (HypoLR), Hypo with Hextend, 6% hydroxyethyl starch in a balanced salt solution (HEX), Hypo with PolyHeme, a polymerized hemoglobin solution (HBOC), or standard resuscitation with LR (StandLR). Animals were bled over 15 min to a mean arterial blood pressure (MAP) of 40 mmHg where the blood pressure (BP) was held for 30 min. Hypo groups were resuscitated to 60 mmHg for 4 h followed by further resuscitation to 80 mmHg. StandLR rats were resuscitated to 80 mmHg immediately after the hemorrhage period. Animals were monitored until death or they were sacrifice at 24 h. Prolonged Hypo with HEX or LR resulted in a trend toward improved 24-h survival compared with C (71%, 65%, and 48%, respectively), and performed at least as well as StandLR (58% survival). HEX required significantly less intravenous fluid (0.7x total estimated blood volume [EBV]) compared with HypoLR (1.9x EBV) and StandLR (3.2x EBV) (P < 0.05). Although HBOC required the smallest fluid volume (0.4x EBV), survival was no better than C and it resulted in the most significant acidosis. These results support the decision to use Hextend for Hypo, a strategy currently being applied on the battlefield. PMID- 15803058 TI - The granulopoietic cytokine response and enhancement of granulopoiesis in mice during endotoxemia. AB - In response to bacterial infection, the production of neutrophils by the bone marrow is accelerated. This study investigated the granulopoietic cytokine response and granulopoiesis during endotoxemia. Male Balb/c mice were intravenously challenged with lipopolysaccharide (LPS, 20 microg in 100 microL of saline per mouse). Control animals received saline alone. In a separate set of experiments, i.v. murine granulocyte colony-stimulating factor (G-CSF; 20 microg/kg) or vehicle (5% dextrose) was administered to mice. Endotoxemia caused a marked increase in G-CSF, keratinocyte-derived chemokine (KC), and macrophage inflammatory protein-2 (MIP-2) in the plasma and bone marrow between 1 and 4 h after the challenge. G-CSF, KC, and MIP-2 mRNA expression was also upregulated in the lung, liver, spleen, and bone marrow between 1 and 4 h after i.v. LPS. Intravenous administration of G-CSF caused a significant increase in G-CSF concentration in the plasma and bone marrow without upregulating G-CSF mRNA expression in the bone marrow. The levels of phospho-signal transducers and activators of transcription 3 and phospho-p44/42 mitogen-activated protein kinase were elevated in bone marrow cells at 30 min and 4 h after i.v. G-CSF and LPS, respectively. Granulocyte-macrophage colony-forming unit counts were significantly increased in the bone marrow, spleen, and blood at 48 h post-i.v. LPS or G-CSF. These data show that extramedullary organs produce granulopoietic cytokines in response to LPS. Because the tissue mass in extramedullary organs far exceeds that in the bone marrow, extramedullary production of these cytokines likely play a critical role in the regulation of the host's granulopoietic response to endotoxemia. PMID- 15803059 TI - Coenzyme q10 confers cardiovascular protection against acute mevinphos intoxication by ameliorating bioenergetic failure and hypoxia in the rostral ventrolateral medulla of the rat. AB - Coenzyme Q10 (CoQ10, ubiquinone) is a highly mobile electron carrier in the mitochondrial respiratory chain that also acts as an antioxidant. We evaluated the cardiovascular protective efficacy of CoQ10 at the rostral ventrolateral medulla (RVLM), a medullary site where sympathetic vasomotor tone originates and where the organophosphate poison mevinphos (Mev) acts to elicit cardiovascular intoxication. Experiments were carried out in adult male Sprague-Dawley rats that were maintained under propofol anesthesia. Microinjection bilaterally of Mev (10 nmol) into the RVLM induced progressive hypotension and minor bradycardia, alongside significant depression of the activity of NADH cytochrome c reductase (enzyme marker for Complexes I and III) or cytochrome c oxidase (enzyme marker for Complex IV) in the mitochondrial respiratory chain, reduction in ATP concentration, or tissue hypoxia in the RVLM. On the other hand, the activity of succinate cytochrome c reductase (enzyme marker for Complexes II and III) remained unaltered. The Mev-induced hypotension, bioenergetic failure, or hypoxia was significantly reversed when CoQ10 (4 microg) was coadministered bilaterally into the RVLM with the organophosphate poison. We conclude that CoQ10 confers cardiovascular protection against acute Mev intoxication by acting on the RVLM, whose neuronal activity is intimately related to the "life-and-death" process. We also showed that amelioration of the selective dysfunction of respiratory enzyme Complexes I and IV in the mitochondrial respiratory chain, the reduced ATP level, and the induced tissue hypoxia in the RVLM are among some of the underlying mechanisms for the elicited protection. PMID- 15803061 TI - The induction of super-resistance using synthetic lipopolysaccharide receptor agonist rescues fatal endotoxemia in rats without excessive immunosuppression. AB - Endotoxin tolerance provides protection against mortality under various conditions of stress. However, the induction of endotoxin tolerance thus far has no clinical application because of endotoxin toxicity and the excessive immune suppression that follows the tolerance induction. In this study, we examined whether a novel, synthetic lipopolysaccharide (LPS) receptor agonist, ER-803058 (ER) can induce endotoxin tolerance with accompanying low toxicity. The stimulative effects of ER on tumor necrosis factor (TNF)-alpha production from RAW264 cells were 50% to 70% lower than those of the corresponding quantities of LPS. ER pretreatment also diminished TNF-alpha secretion induced by a subsequent LPS shock. However, the degree of desensitization with ER pretreatment (10 ng/mL, 55.5% +/- 6.7%; 100 ng/mL, 42.3 +/- 4.9%) was modest in contrast with that measured for the corresponding LPS pretreatment (10 ng/mL, 36.7% +/- 3.7%; 100 ng/mL, 20.0% +/- 3.6%). The minimum in vivo dose (0.02 mg/kg/body weight) of ER induced negligible production of TNF-alpha and interleukin (IL)-6 in rats, and resulted in a modest endotoxin tolerance with respect to TNF-alpha secretion. Although the plasma TNF-alpha level after ER pretreatment was decreased (48.2% +/ 1.1%), the suppression was not statistically significant. Interestingly, even this minimal quantity of ER pretreatment evoked a dramatic improvement in survival (90% survival) against administration of a lethal dose of LPS, which is inconsistent with the modest TNF-alpha suppression. Furthermore, ER pretreatment preserved normal plasma albumin levels and prevented the increase of plasma blood urea nitrogen levels seen with LPS. These results indicate that pretreatment with ER can effectively induce endotoxin tolerance, with a consequent improvement in mortality without toxicity and without subsequent excessive immunosuppression. PMID- 15803060 TI - Coexposure of lead- and lipopolysaccharide-induced liver injury in rats: involvement of nitric oxide-initiated oxidative stress and TNF-alpha. AB - In this study, we investigated the interaction between lipopolysaccharide (LPS) and lead (Pb) and the involvement of tumor necrosis factor-alpha (TNF-alpha) and oxidative stress in Pb-plus-LPS (Pb/LPS)-induced liver damage in rats. Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), TNF alpha, nitric oxide (NO), and lipid peroxidation (LPO) were determined in rats treated with Pb and/or LPS. Pb ranging from 0 to 15 mg/kg dose dependently increased AST, ALT, NO, or LPO in LPS-treated rats. Pretreatment with iNOS inhibitor 1400W reduced NO, LPO, TNF-alpha, AST, and ALT in Pb/LPS-treated rats. Thus, Pb increased LPS-induced liver damage, which might be associated with increased NO-initiated oxidative stress and TNF-alpha in rats. PMID- 15803062 TI - Interleukin 8 dimerization as a mechanism for regulation of neutrophil adherence dependent oxidant production. AB - Interleukin 8 (IL-8), a member of the CXC subfamily of chemoattractant cytokines, induces a range of functional responses in human neutrophils via its interactions with two high-affinity cell-surface receptors, CXCR1 and CXCR2. Like other CXC chemokines, IL-8 forms homodimers at physiologic concentrations. Monomers and dimers bind to CXC receptors with high affinity and induce various functions. Binding to glycosaminoglycans decreases the dimerization constant, enhancing surface-bound dimer formation. However, a specific role for IL-8 dimerization has not been identified. We explored the hypothesis that certain neutrophil responses to IL-8 were induced primarily by the IL-8 dimers. To this end, two dimerization deficient IL-8 mutant proteins, M3 and M4, were used in various functional assays. In contrast to native IL-8, these proteins existed primarily as monomers at micromolar concentrations. The mutants retained high-affinity binding to both CXC receptors and potently induced neutrophil calcium flux, chemotaxis, and elastase release. In contrast to native IL-8, neither mutant inhibited tumor necrosis factor alpha-induced oxidant production. Additionally, M4 was less effective than native IL-8 at desensitizing neutrophil migration. These data suggest that although IL-8 dimers or monomers are sufficient for several neutrophil functions, dimers may participate in suppression of specific surface dependent neutrophil responses. PMID- 15803063 TI - Effects of colloid resuscitation on peripheral microcirculation, hemodynamics, and colloidal osmotic pressure during acute severe hemorrhage in rabbits. AB - We examined the effects of hydroxyethyl starch (HES) on the microcirculation, hemodynamics, and colloidal osmotic pressure in a rabbit model of hemorrhagic shock. A total of 40 rabbits was anesthetized with pentobarbital and isoflurane, and they were mechanically ventilated. An ear chamber was prepared to examine blood vessels by intravital microscopy. Shock was induced by removing nearly half of the circulating blood volume. Twenty rabbits received 20 mL of HES by intravenous infusion immediately after blood letting. Additional HES was then administered intravenously to a total volume of 100 mL. The other 20 rabbits (control) were intravenously given 40 mL of lactated Ringer's solution (LR), followed by additional LR to a total volume of 200 mL, administered under the same conditions as HES. After blood letting, arteriolar diameter decreased similarly in the the HES and LR groups (HES, 40.5% +/- 14.8% of the baseline value versus LR, 43.3% +/- 13.1%). After the completion of infusion, arteriolar diameter significantly recovered to 90.8% +/- 10.2% of the baseline value in the HES group as compared with only 62.6% +/- 10.7% in the LR group (P < 0.005). Recovery of arterial blood flow velocity and blood flow rate was also significantly better in the HES group than in the LR group (P < 0.005). Mean arteriolar pressure, central venous pressure, and plasma colloid osmotic pressure after the completion of infusion were significantly greater in the HES group than in the LR group (P < 0.005). We conclude that intravenous infusion of HES effectively maintains the microcirculation, hemodynamics, and colloidal osmotic pressure in a rabbit model of acute severe hemorrhage. PMID- 15803064 TI - Cardiac function, myocardial glutathione, and matrix metalloproteinase-2 levels in hypoxic newborn pigs reoxygenated by 21%, 50%, or 100% oxygen. AB - After asphyxia, it is standard to resuscitate the newborn with 100% oxygen, which may create a hypoxia-reoxygenation process that may contribute to subsequent myocardial dysfunction. We examined the effects of graded reoxygenation on cardiac function, myocardial glutathione levels, and matrix metalloproteinase (MMP)-2 activity during recovery. Thirty-two piglets (1-3 days old, weighing 1.5 2.1 kg) were anesthetized and instrumented for continuous monitoring of cardiac index, and systemic and pulmonary arterial pressures. After 2 h of hypoxia, piglets were randomized to receive reoxygenation for 1 h with 21%, 50%, or 100% oxygen (n = 8 each), followed by 3 h at 21% oxygen. At 2 h of hypoxemia (PaO2 32 34 mmHg), the animals had hypotension, decreased cardiac index, and elevated pulmonary arterial pressure (P < 0.001 vs. controls). Upon reoxygenation, cardiac function recovered in all groups with higher cardiac index and lower systemic vascular resistance in the 21% group at 30 min of reoxygenation (P < 0.05 vs. controls). Pulmonary artery pressure normalized in an oxygen-dependent fashion (100% = 50% > 21%), despite an immediate recovery of pulmonary vascular resistance in all groups. The hypoxia-reoxygenated (21%-100%) hearts had similarly increased MMP-2 activity and decreased glutathione levels (P < 0.05, 100% vs. controls), which correlated significantly with cardiac index and stroke volume during reoxygenation, and similar features of early myocardial necrosis. In neonatal resuscitation, if used with caution because of a slower resolution of pulmonary hypertension, 21% reoxygenation results in similar cardiac function and early myocardial injury as 50% or 100%. The significance of higher oxidative stress with high oxygen concentration is unknown, at least in the acute recovery period. PMID- 15803065 TI - Unilateral cervical facet dislocation: biomechanics of fixation. AB - STUDY DESIGN: Unilateral facet dislocation was created in human cadaveric cervical spines. Specimens were sequentially instrumented with posterior or anterior screws and plates, and studied biomechanically. OBJECTIVE: To determine the biomechanical differences between anterior and posterior fixation for stabilization of a reduced unilateral cervical facet dislocation. SUMMARY OF BACKGROUND DATA: Although previous studies have compared anterior to posterior instrumentation, no data exist on the biomechanics of either type of stabilization after this particular injury. METHODS: In 6 human cadaveric cervical spine segments, a reproducible unilateral facet dislocation was created and then unlocked (reduced). Nondestructive torques were applied to specimens that were intact, injured-reduced, fixated using posterior nonlocking lateral mass plates, and fixated using a bone graft plus an anterior nonlocking plate. Flexion, extension, lateral bending, and axial rotation were measured stereophotogrammetrically. RESULTS: Lateral mass plating was more effective than anterior plating in limiting motion after reduction of a unilateral facet dislocation. Averaged, over all loading directions, lateral mass plates reduced the range of motion to 17% of normal; anterior plates reduced range of motion to 89% of normal. In all loading directions, lateral mass plates performed significantly better than anterior plates (P < 0.05, paired Student t-tests). CONCLUSIONS: Anterior and posterior plating effectively stabilized a reduced unilateral facet dislocation. Lateral mass fixation provided better immobilization than anterior graft and plate. PMID- 15803066 TI - The influence of posterior instrumentation on adjacent and transfixed facet joints in patients with thoracolumbar spinal injuries: a morphological in vivo study using computerized tomography osteoabsorptiometry. AB - STUDY DESIGN: Subchondral mineralization of adjacent and transfixed facet joints was analyzed in patients with thoracolumbar spinal injuries, both before posterior instrumentation and after removal of the spinal implant. OBJECTIVE: To examine the influence of posterior instrumentation on content and distribution of subchondral mineralization as a correlate of the long-term load acting on the adjacent and transfixed zygapophysial joints. SUMMARY OF BACKGROUND DATA: Posterior stabilization plays an important role in the treatment of spinal injuries and is a standard technique for the treatment of thoracolumbar spinal fractures. Studies have shown that stress and motion in the adjacent segments are altered in the presence of instrumentation. METHODS: Twenty-three patients with thoracolumbar spinal injuries had computerized tomography (CT) during the course of routine posttraumatic diagnostics and subsequently received bisegmental posterior fixation with an internal fixator. Second CT were obtained after removal of the fixation device, which was performed on an average of 9.4 months after the trauma. Patients were divided into 2 groups with follow-up CT taken within either less than 3 months (group A: average 7.3 days, 15 patients) or 6 and more months (group B: average 17 months, 8 patients) after the internal fixator had been removed. Quantitative and qualitative CT osteoabsorptiometry were used to assess changes in subchondral mineralization, reflecting the altered load acting on the adjacent and bridged zygapophysial joints. RESULTS: There was a significant difference between preoperative and postoperative calcium values (P < 0.001) for the whole patient group. Mineralization decrease was significantly more often found than increase (P < 0.001). A separate analysis of the 2 groups of patients revealed significant differences between group A and B (P < 0.001). In group A, a mineralization decrease was found in 61.3% and an increase in 11.0% of the facet joints, while in group B, a mineralization decrease was shown in 21.9% and an increase in 41.0%. No significant differences between adjacent and transfixed facets were found except in group B, in which the suprajacent joints showed a significantly higher mineralization increase than the transfixed joints (P = 0.030). CONCLUSIONS: Decrease in subchondral mineralization indicates reduced load acting on the examined zygapophysial joints. This finding in patients with early follow-up CT seems to be caused by reduced activity in most of the patients until removal of the spinal implant. In patients with longer intervals between removal of the fixator and second CT, higher loads acting on the adjacent and bridged joints are shown morphologically. Whether or not these changes lead to spondylarthritis has to be studied in a long-term follow-up. PMID- 15803067 TI - The influences of Halo-vest fixation and cervical hyperextension on swallowing in healthy volunteers. AB - STUDY DESIGN: Radiographic and electromyographic evaluation of swallowing functions was performed for different positions with a Halo-vest brace. OBJECTIVES: The aim of this study was to clarify the mechanism of dysphagia of cervical hyperextension with a Halo-vest brace in neurologically normal adult volunteers. SUMMARY OF BACKGROUND DATA: Garfin et al reported that 3 of 179 patients had dysphagia attributable to the Halo-vest brace after cervical spinal cord injury. Readjustment of the position of the head in the Halo-vest brace was required in those cases. They concluded that the head-extended position with a Halo-vest brace made it difficult to swallow. However, the details of this dysphagia were not known. METHODS: Six healthy adults volunteers between the ages of 24 and 33 participated in this study. Subjects were radiographically and electromyographically observed swallowing thin liquids at the neutral position without a Halo-vest brace (N-HV), the neutral position with a Halo-vest brace (N+HV) and at hyperextension with a Halo-vest brace (E+HV). RESULTS: In the durational measurements, there were significant changes between the N-HV and E+HV in pharyngeal transit time. The motion measurements showed that the initial hyoid position placed lower from the mandibular plane, and vertical hyoid movement was prolonged in the E+HV. In the electromyographic measurements, greater activity was observed from the suprahyoid muscles in the E+HV. One subject had laryngeal penetration already in the N-HV, and the same subject exhibited aspiration in the E+HV. Another two subjects exhibited penetration in the E+HV. CONCLUSION: The result of this study demonstrated that cervical hyperextension with the Halo-vest brace caused mechanical changes in the swallowing of normal healthy adult volunteers. PMID- 15803068 TI - Clinical accuracy of fluoroscopic computer-assisted pedicle screw fixation: a CT analysis. AB - STUDY DESIGN: Observational cohort study with computerized tomography (CT) analysis of in vivo pedicle screw placement. OBJECTIVE: To evaluate the clinical accuracy of computer-assisted fluoroscopy for the placement of thoracic and lumbosacral (LS) pedicle screws. SUMMARY OF BACKGROUND DATA: Computer-assisted fluoroscopy is an adjunct for the placement of pedicle screws. METHODS: Postoperative CT of 360 titanium pedicle screws (281 LS [L1-S1]; 79 thoracic [T2 T12]) were independently assessed. All screws were placed using the FluoroNav system (Medtronic Surgical Navigation Technologies, Louisville, CO). The relative position of the screw to the pedicle was graded as follows: A = completely in; B = <2 mm breach; C = 2-4 mm breach; and D = >4 mm breach. If an osseous breach occurred, the direction of the breach was further classified. RESULTS: Eight-five percent of screws were completely contained within the pedicle. Fifty-five pedicle breaches (25 medial; 30 lateral) occurred. Pedicle breaches were graded B in 13.1% (47 breaches), C in 1.9% (7), and D in 0.3% (1) of screws. Pedicle breach was significantly higher in the thoracic compared to the LS spine, 31.6% (25/79) and 10.6% (30/281), respectively (P < 0.0001). In the thoracic spine, 72% of pedicle breaches were lateral. In 49% of breaches, the screw diameter was larger than the pedicle diameter. None of the pedicle breaches were associated with neurologic or other clinical sequelae. CONCLUSIONS: The clinical pedicle breach rate in this study is comparable to those reported using conventional techniques with or without fluoroscopic assistance. FluoroNav appears to be a safe adjunct for the placement of thoracic and LS pedicle screws. PMID- 15803069 TI - Surgical management of severe cervical kyphosis with myelopathy in osteogenesis imperfecta: a case report. AB - STUDY DESIGN: A case of a 9-year-old child with Osteogenesis Imperfecta and severe cervical kyphosis associated with wedged vertebrae and progressive neurological deterioration is presented. OBJECTIVE: To highlight the difficulties in surgical management of this condition and to discuss the appropriate surgical approach. SUMMARY OF BACKGROUND DATA: This case demonstrates an unusual case of Osteogenesis Imperfecta with associated wedged vertebrae causing a quadriparesis. Surgical decompression and stabilization can be performed with resolution of symptoms even in this age group with the appropriate approach and implants. METHODS: A 9-year-old girl presented with progressive cervical kyphosis and quadriparesis. At the age of 3 years, she underwent posterior cervical fusion (C1 C6) for instability. Radiological and laboratory investigations confirmed the diagnosis of Osteogenesis Imperfecta, and radiographs of the cervical spine revealed a kyphotic deformity of 120 degrees . Magnetic resonance imaging and computerized tomography scans showed anterior cord compression attributable to wedged vertebrae at C3 and C4. Magnetic resonance imaging-angiography was performed before surgery to identify the anatomic position of the vertebral arteries. A modified anterolateral approach to the upper cervical spine was performed, and anterior C3 and C4 corpectomies with interbody cage and plate fixation were carried out. RESULTS: After surgery the patient made a full neurological recovery, and significant correction of the deformity was achieved and maintained at follow-up. CONCLUSIONS: Cervical kyphotic deformity in Osteogenesis Imperfecta is uncommon. Association of this condition with wedged vertebrae is rare. Surgical decompression of the upper cervical spine is a challenging problem in the presence of this deformity. Which surgical approach to use is controversial. There are difficulties exposing wedged vertebrae by a standard anterior approach, and hence we have used a modified anterolateral approach to address this surgical problem, because a posterolateral approach was impossible with the intervening vertebral arteries. Spinal stabilization in children with Osteogenesis Imperfecta and poor bone stock is a challenge. We have used a small diameter MOSS cage ("Harms mesh cage") with maxillofacial plate and screws to achieve stabilization and fusion. PMID- 15803071 TI - A randomized clinical trial comparing two physiotherapy interventions for chronic low back pain. AB - STUDY DESIGN: A randomized clinical trial with blinded assessment. OBJECTIVES: To investigate the clinical efficacy of 2 active interventions for patients with chronic low back pain. SUMMARY OF BACKGROUND DATA: Manual therapy and exercise prescription are treatments frequently prescribed for patients with chronic low back pain. The evidence for the relative benefit of these treatments is limited, and questions concerning the most appropriate type of intervention remain unanswered. METHODS: Eighty patients with chronic low back pain (>3 months) were randomized to one of the following treatments, involving 8 treatments over 8 weeks; 1) one-to-one treatment involving 30 minutes of manual therapy (mobilizations to the spine) and spinal stabilization exercises, and 2) a 10 station exercise class involving aerobic exercises, spinal stabilization exercises, and manual therapy. Three physiotherapists led the hour long group with a maximum of 10 patients. Questionnaires were completed, and physical measurements were taken by a blinded observer before randomization, at the completion of treatment, and at 6 months and 12 months after the completion of treatment. The intention-to-treat principle was used in data analysis. RESULTS: Eleven patients dropped out of the individual treatment sessions and 7 dropped out of the exercise group. There was a significant reduction (reduced disability) in the questionnaire score in both groups, and there were significant increases in range for all the physical movements tested in both groups. The exercise group was 40% more cost effective than the individual treatments. CONCLUSION: Both forms of intervention were associated with significant improvement. On-going clinical research is necessary to provide guidance as to the clinical efficacy of various forms of intervention. PMID- 15803072 TI - Differential effects of fibronectin fragment on proteoglycan metabolism by intervertebral disc cells: a comparison with articular chondrocytes. AB - STUDY DESIGN: This in vitro study used the alginate bead culture system to probe for differences in the effects of fibronectin fragment on cell proliferation and proteoglycan metabolism by different populations of intervertebral disc cells and articular chondrocytes. OBJECTIVE: To compare the effects of fibronectin fragment on cell proliferation, and proteoglycan synthesis and degradation by cells from the nucleus pulposus, the anulus fibrosus, and articular cartilage. SUMMARY OF BACKGROUND DATA: In articular cartilage, the administration of fibronectin fragment stimulates cartilage degeneration. Fibronectin fragment levels were increased in human intervertebral discs with increased disc degeneration. Fibronectin fragment injected into the central region of the rabbit intervertebral disc induced a progressive degeneration of that disc. METHODS: Bovine tails and metacarpophalangeal joints from 14- to 18-month-old animals were used. Alginate beads containing cells isolated from intervertebral discs and articular cartilage were cultured with (1-100 nmol/L) or without (control) fibronectin fragment in the presence of 10% fetal bovine serum. In these cultures, deoxyribonucleic acid and proteoglycan contents, as well as the rate of proteoglycan synthesis were determined. Proteoglycan degradation was measured in cultures with or without 10 nmol/L fibronectin fragment. RESULTS: In articular chondrocytes, fibronectin fragment strongly suppressed proteoglycan synthesis and stimulated proteoglycan degradation; the total proteoglycan content was diminished in a dose-dependent manner. Compared to articular chondrocytes, nucleus pulposus cells responded to fibronectin fragments in a similar, although less pronounced manner. On the other hand, anulus fibrosus cells treated with fibronectin fragment did not show any significant effects on proteoglycan degradation. A slight but statistically significant up-regulation of proteoglycan synthesis was observed at 10 nmol/L fibronectin fragment in outer anulus fibrosus cells. However, total proteoglycan content was decreased significantly at high concentrations of fibronectin fragment. CONCLUSIONS: Fibronectin fragment has different effects on cell proliferation, proteoglycan synthesis, degradation, and accumulation by articular chondrocytes and intervertebral disc cells. The different effects of fibronectin fragment in those different cell types suggest metabolic differences between these cells, and may further suggest differences in pathways of fibronectin fragment signaling as well as a differential need of these cells to be involved in tissue remodeling in which both anabolic and catabolic pathways might be altered. PMID- 15803073 TI - Functional impairment as a predictor of spine loading. AB - STUDY DESIGN: Spine loadings during a variety of lifting exertions were compared with individual torso kinematic abilities. Relationships were evaluated between these measures. OBJECTIVE: To determine if trunk kinematic status (functional impairment) is indicative of spine loading increases in patients with low back pain (LBP) compared to asymptomatic individuals. SUMMARY OF BACKGROUND DATA: Recurrent LBP is a common and costly problem that may be related to increased spine loads in those individuals with LBP. Previous studies suggest that patients with LBP had greater loading than their asymptomatic counterparts when performing work. However, we know little about how to identify when a patient with LBP can resume lifting tasks without having exaggerated spine loading. METHODS: Sixty-two patients with LBP and 61 who were asymptomatic were evaluated for signs of kinematic compromise (i.e., inability to generate normal trunk kinematic patterns) during a prelift test. All subjects were then asked to perform a variety of lifting exertions that varied in lift origin (region), lift asymmetry position, and weight lifted. An electromyography-assisted model was used to evaluate spine loading in each subject during the lifting exertions. Statistical models were used to assess the relationship between kinematic compromise and spine loading. RESULTS: Patients with LBP had greater spine loading as well as greater kinematic compromise. The degree of kinematic compromise was related to the degree of spine loading increases in those individuals with LBP. A statistical model was developed that was able to describe 87% of the variability in compression, 61% in anteroposterior shear, and 65% in lateral shear. CONCLUSIONS: Those patients with greater kinematic compromise used higher levels of antagonistic muscle coactivation that not only reduced trunk motion but also resulted in increases in spine loading. Given the degree of kinematic compromise and the lifting task conditions, a method has been devised to predict the increase in spine loading above and beyond that of an asymptomatic individual when performing typical materials handling tasks. PMID- 15803074 TI - Effect of total disc replacement with ProDisc on intersegmental rotation of the lumbar spine. AB - STUDY DESIGN: The mechanical behavior of the lumbar spine after insertion of a ProDisc prosthesis was studied using 3-dimensional nonlinear finite element models. OBJECTIVE: To determine how the mechanical behavior of the lumbar spine is affected by the implant position and height, as well as by removing different portions of the natural disc and resuturing the anterior longitudinal ligament (ALL). SUMMARY OF BACKGROUND DATA: Little is known about how the affected and adjacent levels of the spine are influenced by the implant height and position or by the surgical procedure. METHODS: The artificial disc ProDisc was integrated in a validated 3-dimensional, nonlinear, finite element model of the lumbar spine. The model was loaded with the upper body weight and muscle forces to simulate standing, 30 degrees flexion, 15 degrees extension, and 6 degrees axial rotation. The disc position was varied by up to 2 mm in both an anterior and posterior direction. Three different disc heights were investigated, as well as the influence of removing different portions of the natural disc and resuturing the ALL. RESULTS: Implant position strongly influences intersegmental rotation for the loading cases of standing and flexion. A disc height 2 mm in excess of the normal disc space increases intersegmental rotation at implant level during standing and extension. The values for intersegmental rotation are closer to those for the intact spine when lateral portions of the anulus are not removed. Resuturing the ALL has a strong effect on the loading cases of extension and standing. CONCLUSIONS: When implanting an artificial disc, great care should be taken in choosing the optimal height and correct position for the implant. Lateral portions of the anulus should be preserved whenever possible. A perfect reconstruction of the ALL would help to restore the biomechanics to normal. PMID- 15803075 TI - The treatment mechanism of an interspinous process implant for lumbar neurogenic intermittent claudication. AB - STUDY DESIGN: The spinal canal and neural foramina dimensions of cadaver lumbar spines were quantified during flexion and extension using magnetic resonance imaging before and after placement of an interspinous process implant. OBJECTIVE: To quantify the effect of the implant on the dimensions of the spinal canal and neural foramina during flexion and extension. SUMMARY OF THE BACKGROUND DATA: Lumbar neurogenic intermittent claudication symptoms are typically exacerbated during extension and relieved during flexion. It is understood that the dimensions of the spinal canal and neural foramen increase in flexion and decrease in extension. The authors hypothesized that an interspinous process implant would significantly prevent narrowing of the canal and foramina in extension and have no significant effect in flexion. METHODS: Eight L2-L5 specimens were positioned to 15 degrees of flexion and 15 degrees of extension using a positioning frame. Each specimen was magnetic resonance imaged with and without an interspinous implant (X STOP) placed between the L3-L4 spinous processes. Canal and foramina dimensions were compared between the intact and implanted specimens using a repeated measures analysis of variance with a level of significance of 0.05. RESULTS: In extension, the implant significantly increased the canal area by 18% (231-273 mm), the subarticular diameter by 50% (2.5-3.7 mm), the canal diameter by 10% (17.8-19.5 mm), the foraminal area by 25% (106-133 mm), and the foraminal width by 41% (3.4-4.8 mm). CONCLUSIONS: The results of this study show that the X STOP interspinous process implant prevents narrowing of the spinal canal and foramina in extension. PMID- 15803076 TI - Effects of wearing halo vest on gait: three-dimensional analysis in healthy subjects. AB - STUDY DESIGN: The effects of a halo vest on the gait were studied. The motions of the head, shoulder girdle, trunk, and hip were analyzed with the vest and tong either connected by bars or unconnected. OBJECTIVE: To analyze effects of wearing a halo vest using three-dimensional motion analysis. SUMMARY OF BACKGROUND DATA: We have little information on the effects of halo vests on gait. METHODS: Twenty eight healthy male volunteers participated (age, 32 +/- 7.2; average +/- SD). The halo tong and vest were applied in a noninvasive way. Gait patterns and three dimensional motions of the head, shoulder girdle, trunk, pelvis, and hip in control and halo vest gaits were analyzed using rectangular force plates and a VICON system (Vicon Motion System Ltd., Oxford, UK). RESULTS: The halo vest decreased the gait speed; it increased the duration of strides and decreased the length of strides. It also decreased the motions between the shoulder girdle and the trunk, the trunk and the pelvis, and decreased the motion of the hip in 3 planes. CONCLUSION: Our study provides preliminary evidence that halo vests increase stride time and decrease stride length and reduce motions between the shoulder girdle and the trunk, the trunk and the pelvis, and decreases the motion of the hip. PMID- 15803077 TI - Long-term outcome of laminoplasty for cervical myelopathy due to disc herniation: a comparative study of laminoplasty and anterior spinal fusion. AB - STUDY DESIGN: A retrospective study was conducted. OBJECTIVE: To compare the long term outcomes after laminoplasty and anterior spinal fusion (ASF) for cervical myelopathy secondary to disc herniation. SUMMARY OF BACKGROUND DATA: There have been no reports of long-term comparative studies of laminoplasty and ASF for cervical myelopathy due to disc herniation. METHODS: Of 21 patients who underwent ASF only between 1984 and 1987, 15 were followed up. Of 22 patients who underwent laminoplasty only between 1987 and 1994, 18 were followed up. There were no significant differences in preoperative prognostic factors between the 2 groups. Average follow-up was 15 years in the ASF group and 10 years in the laminoplasty group. Neurologic and radiologic results were examined. RESULTS: Laminoplasty and ASF provided equal neurologic improvement. In the ASF group, additional surgery was required for bone graft complications in 2 patients and for adjacent spondylosis in 1. In the laminoplasty group, one patient had C5 palsy, and intractable axial pain developed in 5 patients after surgery, but no patients needed additional surgery. CONCLUSIONS: Because the 2 procedures provided the same neurologic improvement, the risks of bone graft complication with ASF must be weighed against the risks of chronic neck pain associated with laminoplasty for determining the best technique. Therefore, because our present surgical strategy for cervical myelopathy due to disc herniation, laminoplasty is the procedure of choice except for a patient with single level disc herniation without developmental canal stenosis, who is considered to be a good candidate for ASF. PMID- 15803078 TI - Looking away from whiplash: effect of head rotation in rear impacts. AB - STUDY DESIGN: Twenty healthy volunteers in a laboratory were subjected to rear end impacts 4.4, 7.9, 10.9, and 13.1 m/s acceleration, with head rotation to the right and left. OBJECTIVE: The purpose of this study was to determine the response of the cervical muscles to increasing low-velocity rear impacts when the head is rotated at the time of impact. SUMMARY OF BACKGROUND DATA: A previous study of rear impacts with head in neutral posture suggests that the burden of impact is borne primarily by the sternocleidomastoid muscles bilaterally. To improve automobile designs to prevent whiplash injury, we need to understand the response of the cervical muscles to whiplash-type perturbations in less-than ideal conditions, such as when the head is rotated to the right and left at the time of rear-end impact. METHODS: Triaxial accelerometers recorded the acceleration of the sled, torso at the shoulder level, and head of the participant, while bilateral electromyograms of the sternocleidomastoids, trapezii, and splenii capitis were also recorded. RESULTS: For participants having a rear-end impact, whether having the head rotated to the left or right at the time of impact, the muscle responses increased with increasing levels of acceleration (P < 0.01). The time to onset and time to peak electromyogram for all muscles progressively decreased with increasing levels of acceleration (P < 0.01). Which muscle responded most to a whiplash-type neck perturbation was determined by the direction of head rotation. With the head rotated to the left, the right sternocleidomastoid generated 88% of the maximal voluntary contraction electromyogram (at least triple the response of other muscles). In comparison, the left sternocleidomastoid, both trapezii, and the splenii capitis generated on average only 10% to 30% of the maximal voluntary contraction electromyogram with head rotated to the left. On the other hand, with the head rotated to the right, the left sternocleidomastoid generated 94% of the maximal voluntary contraction electromyogram (again, at least triple the response of other muscles). CONCLUSIONS: If the head is rotated out of neutral posture at the time of rear impact, the injury risk tends to be greater for the sternocleidomastoid muscle contralateral to the side of rotation. Measures to prevent whiplash injury may have to account for the asymmetric response because many victims of whiplash are expected to be looking to the left or right at the time of collision. PMID- 15803079 TI - Ewing's sarcoma of the mobile spine. AB - STUDY DESIGN: A retrospective analysis was performed. OBJECTIVES: To determine the oncological outcome of patients with nonmetastatic Ewing's sarcoma of the mobile spine treated with systemic multiagent chemotherapy combined with radiation therapy for definitive local control. SUMMARY OF BACKGROUND DATA: To our knowledge, there are no studies that evaluate the oncological outcome of patients with nonmetastatic Ewing's sarcoma of the mobile spine treated with systemic chemotherapy and radiation therapy for definitive local control. METHODS: Thirteen patients with nonmetastatic Ewing's sarcoma of the mobile spine were treated with high-dose multiagent chemotherapy combined with radiation therapy for definitive local control from 1971 to 2000 at a single institution. Patients were observed for a minimum of 2 years or until death. Neurological function, local recurrence, distant relapse, and treatment-related complications were evaluated. RESULTS: There were 8 females and 5 males with a mean age of 19 years (ranging from 7-26 years). The mean follow-up time was 65 months (median 28 months; ranging from 2 to 218 months). All patients presented with pain. Motor deficits were present in 6 patients. Ten patients had a decompressive laminectomy. Improved pain control, as determined by narcotic use, was noted in 12 (92%) patients. Ten patients maintained or improved motor function by at least 1 Frankel grade, while 3 had deterioration of motor function. The disease-free survival rate was 49% and 36% at 5 and 10 years. Five (38%) patients were free of disease at last follow-up. Seven patients developed metastatic disease. Three (23%) patients developed a local recurrence. One of these patients had paraplegia associated with the local recurrence. Five patients developed 8 treatment-related complications. Four of the 10 (40%) patients that had a laminectomy developed progressive kyphosis. Two of these patients also developed late-onset cauda equina syndrome along with the deformity. One of these patients also developed cardiomyopathy associated with adriamycin cardiotoxicity. One patient developed a nonhealing pressure ulcerover a prominent spinous process. CONCLUSIONS: The current study provides historical data on a relatively homogeneous group of patients withEwing's sarcoma of the mobile spine treated with multiagent chemotherapy combined with radiation therapy for definitive local control. Systemic chemotherapy combined with current spinal resection and reconstruction techniques may lead to improved oncological and clinical outcomes. PMID- 15803080 TI - Spinal exostoses: analysis of twelve cases and review of the literature. AB - STUDY DESIGN: Retrospective review of spinal exostoses treated at our institution and literature review. OBJECTIVES: Review of 12 cases of spinal exostoses treated at our institution compared with 165 cases of spinal exostoses reported in the literature. SUMMARY OF BACKGROUND DATA: Spinal exostoses are uncommon. Most reports consist of 1 to 3 cases. The relationship between solitary exostoses and those associated with multiple hereditary exostoses (MHE), as well as the incidence of intraspinal and extraspinal location, symptoms presentation, and results of treatment are unclear. METHODS: The medical records, operative reports, and diagnostic imaging of 12 patients with spinal exostoses treated at our institution between 1972 and 2002 were reviewed. The literature was reviewed using MEDLINE search of English literature and bibliographies of published manuscripts. RESULTS: Solitary spinal exostoses were more common than those associated with MHE. Lesions were most common in the upper cervical spine and originated from the posterior elements. Patients with exostoses associated with MHE were significantly younger and had a higher incidence of symptoms consistent with neural structure compression than patients with solitary exostoses. Complete excision resulted in resolution of preoperative symptoms. Intralesional excision resulted in recurrence in all cases. CONCLUSIONS: Spinal exostoses are more common than reported previously. Patients with MHE that present with back pain or neurological symptoms should produce a high index of suspicion. Evaluation should include both computed tomography and magnetic resonance imaging to define the origin of the exostosis and the presence of neural structure compression. Surgical excision should be preformed en bloc. PMID- 15803081 TI - Histological evaluation of biopsies obtained from vertebral compression fractures: unsuspected myeloma and osteomalacia. AB - STUDY DESIGN: A histological evaluation of biopsies obtained from presumed osteoporotic vertebral compression fractures (VCF) to confirm possible osteomalacia after tetracycline labeling. OBJECTIVE: To describe the results of a series of biopsies obtained at the time of vertebral augmentation in presumed osteoporotic VCF, with special reference to the presence of unmineralized bone (osteomalacia) and occult or unconfirmed plasma cell dyscrasia. SUMMARY OF BACKGROUND DATA: Vertebral augmentation is now widely performed as a method to treat osteoporotic or osteolytic VCF. However, the influence of underlying pathology on the effect of treatment is unclear. METHODS: As of October 2003, 178 biopsies were obtained from 142 patients with VCF during 246 kyphoplasty procedures. There were 110 one-level, 28 two-level, and 4 three-level biopsies. Patients included 41 men and 101 women, with an average age of 72 years (range 40 90). The patients consented to this procedure, and 25 received tetracycline (1g/day, in 2 doses separated by 6 days). Vertebral body biopsies were taken using a trephine just before the kyphoplasty procedure. The biopsies were fixed, embedded, and stained with toluidine blue and hematoxylin eosin, and were viewed with transmitted light. Unstained sections were viewed under fluorescent light to detect tetracycline labels. RESULTS: The 178 biopsy levels included: T4 (3), T5 (1), T6 (4), T7 (13), T8 (12), T9 (8), T10 (11), T11 (17), T12 (28), L1 (25), L2 (14), L3 (13), L4 (17), and L5 (12). All specimens showed fragmented bone with variable amounts of unmineralized bone (osteoid), suggesting bone remodeling and/or fracture healing. Woven bone and cartilaginous tissue were often present, representing fracture callus formation. The biopsies obtained from 30 patients (21%), including 4 who received tetracycline, showed significantly increased osteoid, suggesting either increased bone remodeling activity or mineralization defect (osteomalacia). One sample from these 4 patients who received tetracycline showed no tetracycline labels, essentially diagnostic of osteomalacia. The biopsies also provided definitive diagnoses for one case of unsuspected and 3 cases of unconfirmed plasma cell dyscrasia. CONCLUSIONS: The majority of biopsies from this series of patients revealed findings consistent with various stages of fracture healing. Osteoid seams were increased in 30 patients, representing either increased bone remodeling or osteomalacia. More cases with tetracycline labeling will help elucidate the true incidence of osteomalacia in this population. As we confirmed 4 cases of plasma cell dyscrasia, we advocate a biopsy during each first-time vertebral augmentation procedure. PMID- 15803082 TI - Correlation between neurological deficit and spinal canal compromise in 198 patients with thoracolumbar and lumbar fractures. AB - STUDY DESIGN: Review the correlation between loss of spinal canal diameter and neurological deficits in patients with thoracolumbar and lumbar burst fractures. OBJECTIVES: To try to establish a correlation between neurological deficits and the degree of narrowing of the spinal canal. SUMMARY OF BACKGROUND DATA: The correlation between the degree of protrusion of the bone fragments into the spinal canal and the incidence of neurological deficits in thoracolumbar and lumbar burst fractures has not been well established, raising a lot of controversies in the literature. METHODS: Manual measurements of axial-computed tomography scan films using a millimetric ruler were made from the spinal canal of 198 patients admitted to the hospital with thoracolumbar and lumbar burst fractures, from 1989 to 2000. RESULTS: The probability of neurological deficit in a patient with 25, 50, and 75% narrowing of the thoracolumbar spinal canal was found to be 12, 41, and 78%, and in the lumbar spinal canal it was 8, 30, and 68%, respectively. CONCLUSIONS: The narrowing of the spinal canal proved to be a predictive factor in establishing early neurological deficits in thoracolumbar and lumbar burst fractures, showing a positive correlation between narrowing of the spinal canal and the severity of the incomplete neurological deficit by the Frankel classification. Patients with complete spinal cord injuries (Frankel A) did not show this correlation. PMID- 15803083 TI - Standardized provocation of lumbar spine mobility: three methods compared by radiostereometric analysis. AB - STUDY DESIGN: By radiostereometric analysis (RSA), the intervertebral mobility was measured for the 3 most distal lumbar disc levels in 12 patients at 3 types of standardized provocation: changing body position from supine to standing, supine to standing with load, and supine to sitting. OBJECTIVE: To study the validity and intervertebral mobility response to 3 types of standardized provocation of the lower lumbar spine. SUMMARY OF BACKGROUND DATA: Active flexion extension is used in clinical practice to detect pathologic kinematics in the lumbar spine. However, the actual mobility response is affected by variations in pain level and muscular spasm. Standardizing the examinations reduces these confounding factors, and brings the possibility to study intervertebral mobility over time and between individuals. A standardized method using traction compression was introduced earlier but questioned due to the insufficient mobility provided. METHODS: After percutaneous application of tantalum indicators for RSA, the intervertebral mobility was measured for the 3 most distal lumbar disc levels in 12 adult patients with low back pain and disc degenerative disease. Each patient was examined in 4 separate positions: supine, normal standing, standing with sandbags (total weight 20 kg) on the shoulders, and sitting. The translatory movements induced by the patient changing position from supine to the other 3 positions were measured and the mobility response compared for the 3 types of provocation. RESULTS: The sagittal mobility induced by the sitting provocation was significantly higher than both the other types of provocation on all the 3 spinal levels measured. The same result was seen for the vertical translations except for the L3-L4 level, where the standing with load provocation was equal to sitting. Transverse translations were small and did not differ for the 3 types of provocation. CONCLUSION: The intervertebral mobility response provided in the lumbar spine when changing position from supine to sitting is more pronounced than both to standing and to standing with a 20 kg load. By the supine to sitting provocation, mean lumbar mobility is increased, and mobility can be revealed in some cases being stable in the other 2 situations. Combined with RSA, the method brings a refined possibility for studying lumbar spine kinematics. PMID- 15803084 TI - An epidemiologic study of MRI and low back pain in 13-year-old children. AB - STUDY DESIGN: Cross-sectional cohort study of a general population. OBJECTIVE: To describe associations between "abnormal" lumbar magnetic resonance imaging (MRI) findings and low back pain (LBP) in 13-year old children. SUMMARY AND BACKGROUND DATA: Very little is known about the distribution of lumbar MRI findings and how they are associated with LBP in youngsters. METHODS: Disc abnormalities, as well as nerve root compromise, endplate changes, and anterolisthesis were identified from MRI studies of 439 children. LBP was identified from structured interviews. Associations are presented as odds ratios (OR). RESULTS: Signs of disc degeneration were noted in approximately 1/3 of the subjects. Reduced signal intensity and irregular nucleus shape in the upper 3 lumbar discs were significantly associated with LBP within the last month (OR, 2.5-3.6), whereas reduced signal intensity and disc protrusion at L5-NS1 were associated with seeking care (OR, 2.8 and 7.7, respectively). Endplate changes in relation to the L3 discs were associated with LBP month and seeking care (OR, between 9.7 and 22.2). Anterolisthesis at L5 was associated with seeking care (OR, 4.3). There were obvious differences between genders: degenerative disc changes in the upper lumbar spine were more strongly associated with LBP in boys, while disc abnormalities in the lower lumbar spine were more strongly associated with seeking care in girls. CONCLUSIONS: In children, degenerative disc findings are relatively common, and some are associated with LBP. There appears to be a gender difference. Disc protrusions, endplate changes, and anterolisthesis in the lumbar spine were strongly associated with seeking care for LBP. PMID- 15803085 TI - Evaluating MRI as a technique for visualizing the neurocentral junction. AB - STUDY DESIGN: Magnetic resonance imaging (MRI) of the neurocentral junction (NCJ) was correlated with gross anatomic and histologic sections acquired at the same position. OBJECTIVES: To determine the composition of the MRI of the NCJ and to explore the long-standing discrepancy between the ages of closure of the NCJ as determined by anatomic and imaging studies. SUMMARY OF BACKGROUND DATA: Disparate growth at the site of the NCJ has been implicated as a potential cause of adolescent idiopathic scoliosis since the early 1900s. Although anatomic studies have refuted this theory by maintaining that the NCJ closes before the age of 10 years, recent MRI studies have suggested that the NCJ remains open until adolescence. METHODS: One hundred fourteen porcine NCJs in various stages of development were visualized using MRI. Gross anatomic and histologic sections were acquired at the same position as MRI for correlation. RESULTS: The presence or absence of NCJ cartilage was represented by the presence or absence of a line on MRI. However, the NCJ image overestimated the width of the NCJ cartilage in the anteroposterior direction during development. As the NCJ underwent the process of closure, the NCJ image underestimated the extent of closure in the medial-lateral direction. CONCLUSIONS: Although MRI underestimated the extent of NCJ closure, MRI accurately showed whether cartilage was present or absent at the NCJ site. MRI determination of the age of NCJ closure appears reliable, and further exploration of the asymmetric growth hypothesis is required. PMID- 15803086 TI - A systematic review of sociodemographic, physical, and psychological predictors of multidisciplinary rehabilitation-or, back school treatment outcome in patients with chronic low back pain. AB - STUDY DESIGN: A systematic review. OBJECTIVE: To determine predictors of outcome of multidisciplinary rehabilitation-or back school treatment for patients with chronic low back pain. SUMMARY OF BACKGROUND DATA: Numerous reviews have been performed to gain insight into which patients benefit from which treatment. However, no review has systematically focused on predictors from multiple domains (i.e., sociodemographic, physical, and psychological), or on treatment outcome measured as activity limitation or participation restriction. METHODS: Studies were found by searching medical and psychological databases, and screening references. Two reviewers independently assessed the methodological quality using standard criteria. Studies were only included if they met a predefined level of internal validity. A qualitative analysis was performed. RESULTS: Heterogeneity among studies in patient characteristics, predictors, treatment, and outcomes limited evidence. All reviewed studies were descriptive or exploratory in nature. Consistent evidence was found for the predictive value of pain intensity (more pain--> worse outcome), several work-related parameters (e.g., high satisfaction- > better outcome), and coping style (less active coping--> better outcome). Other sociodemographic and physical variables consistently lacked predictive value. No consistent evidence was found for other psychological variables. CONCLUSIONS: It is impossible to define a generic set of predictors of outcome of multidisciplinary rehabilitation and back schools for patients with chronic low back pain because the reviewed studies were descriptive or exploratory in nature, and most predictors were only studied once. Nevertheless, for several predictors, consistent evidence was found. Large confirmatory studies are needed to test the value of these predictors. PMID- 15803087 TI - Lumbar disc herniation: evaluation of information on the internet. AB - STUDY DESIGN: An original study was performed evaluating the information presented on existing web sites for the topic of lumbar disc herniation. OBJECTIVES: The purpose of this study was to evaluate the type and quality of internet information available to patients on the topic of lumbar disc herniation. Our secondary objectives were to rank the identified World Wide Web sites with respect to the caliber of relevant information and to determine the propensity for secondary commercial gain by the web site sponsors. SUMMARY OF BACKGROUND DATA: Two-thirds of the United States population "surfs" the internet. A substantial percentage of internet users search for medical information on the World Wide Web. Because no standards exist regarding the publication of medical literature on the internet, the relevant web sites vary dramatically in terms of content and quality. Misleading or inaccurate information poses a theoretical risk to patients seeking treatment for medical conditions. METHODS.: Five search terms (lumbar disc herniation, herniated nucleus pulposus, herniated disc, slipped disc, and sciatica) were entered into 5 commonly used search engines. The first 25 links displayed by each engine were evaluated for a theoretical total of 625 web sites. Each site was evaluated in terms of content, authorship, and secondary commercial gain. An information quality score of 0 to 25 points was generated for each site; a score of 20 or greater was indicative of "high quality" content. RESULTS: Our search identified 169 unique web sites of which only 16 (9.5%) scored >or=20 on the information quality score; 103 (60.9%) scored or=150 micromol/L. Only 15% children and 26% adults had HbA1c<7%, 86.2% children and 62.7% adults had blood pressure<130/85 mmHg; 58% children and 80% adults had at least 3 daily insulin injections. In adults, the risk of experiencing at least one complication was linked significantly with diabetes duration, HbA1c, and socio-economic status. Age, sex, type of insulin therapy, tobacco consumption, and blood pressure control were not significant parameters. Ketoacidosis in the preceding year was only linked with HbA1C and socio-economic status. CONCLUSION: Although this sample of patients had overall a fair socio-economic status and were followed-up by specialists of diabetes care, metabolic and blood pressure control were not optimal. The care of French type 1 diabetics could probably be improved by a stricter control of glycaemia and blood pressure, and an earlier use of intensive insulin treatment, with a particular focus on adolescents and patients with the lowest socio-economic status. PMID- 15803113 TI - Fas/Fas-Ligand pathway is impaired in patients with type 2 diabetes. Influence of hypertension and insulin resistance. AB - OBJECTIVES: In type 2 diabetic patients with no cardiac history or symptoms, 1) to evaluate whether the soluble forms of Fas (sFas) and Fas-ligand (sFasL), involved in apoptosis, may be markers of silent coronary disease or related to hypertension or microangiopathic complications; 2) to examine the effect of short term glycemic control on sFas and sFasL. METHODS: (1) sFas and sFasL were measured with the ELISA method in 44 asymptomatic diabetic patients, 33 with hypertension, and with a normal myocardial scintigraphy (n=14), with silent myocardial ischemia (SMI) and without (n=15) or with (n=15) significant coronary stenoses; and in 14 controls; (2) sFas and sFasL were measured in 15 poorly controlled diabetic patients before and after 7 days of CSII treatment. RESULTS: (1) sFas and sFasL differed in the four groups of patients (p=0.003 each). sFas was significantly higher in the patients with SMI without (p=0.035) and with coronary stenoses (p=0.002) than in the control group. sFasL was lower in the three groups of diabetic patients (p<0.05 each) than in control group. In the diabetic population, sFas correlated positively with hypertension (p=0.021), and sFasL negatively with hypertension (p=0.027) and HOMA index in the non-insulin treated patients (p=0.049); (2) sFas did not differ before or after CSII, and there was a marginal decrease in sFasL. CONCLUSION: Fas-mediated apoptosis is involved in type 2 diabetes and might be associated with hypertension and/or its vascular consequences. sFasL might be affected by insulin resistance. sFas and sFasL are not effective markers of SMI. PMID- 15803114 TI - Low-grade systemic inflammation, hypoadiponectinemia and a high concentration of leptin are present in very young obese children, and correlate with metabolic syndrome. AB - OBJECTIVE: To determine the concentration levels of C-reactive protein (CRP), leptin and adiponectin in obese pre-pubertal children, and their possible relation with metabolic syndrome, fibrinogen and plasminogen activator inhibitor 1. METHODS: A study was carried out in 51 obese children (aged 6 to 9 years) and the same number of non-obese children (control group), matched by age and sex. (Cross-sectional study of obese children). Body mass index (BMI), waist/hip ratio (WHR) and blood pressure were determined for each child. Serum CRP, leptin, adiponectin, glucose, insulin, lipid profile, plasminogen activator inhibitor-1 (PAI-1) and fibrinogen were all measured. RESULTS: The levels of CRP serum (1.67+/-0.222 vs 0.92+/-0.16 mg/l) and leptin (15.56+/-1.27 vs 4.68+/-0.62 ng/ml) were significantly higher in obese children. The adiponectin level was significantly higher in non-obese children (11.58+/-0.63 vs 9.64+/-0.49 microg/dl). In the obese group, log. CRP showed a positive correlation with BMI, insulin, homeostasis model assessment (HOMA), triglycerides, alanine aminotransferase (ALT), uric acid, PAI-1, fibrinogen and interleukin 6 (IL-6), and correlated negatively with apolipoprotein A-I and high-density lipoprotein cholesterol (HDL-C). The leptin was positively correlated with BMI, insulin, HOMA, triglycerides and PAI-1 and negatively with Apo A-I and HDL-C. Adiponectin correlated negatively with BMI, insulin, HOMA, and triglycerides. CONCLUSIONS: Low-grade systemic inflammation, elevated leptin concentration and low adiponectin level are described in very young obese children, correlating with a range of variables of metabolic syndrome. Inflammation and adipocytokines can play an important role in the etiopathogeny of metabolic syndrome. PMID- 15803115 TI - Increased plasma concentration of nitric oxide in type 2 diabetes but not in nondiabetic individuals with insulin resistance. AB - OBJECTIVE: Insulin resistance (IR) is a key element in the pathogenesis of type 2 diabetes. The results of recent experiments on insulin-mediated vasodilatation have suggested that vascular insensitivity is a component of IR. However, it is still controversial that patients with type 2 diabetes have a decreased ability of insulin to increase endothelial nitric oxide (NO) release. METHOD: Plasma concentration of NO was examined in 26 patients with type 2 diabetes and 78 nondiabetic volunteers during an insulin suppression test. The test measured the efficacy of insulin in promoting disposal of the infused glucose load, in which the steady state plasma glucose (SSPG) during the 150-180 min of the test was used as an index of IR. Plasma NO levels were assayed by measurement of the stable end products of their metabolism. Comparison of plasma NO levels between groups were performed by Mann-Whitney test and relationships between SSPG and different variables were analyzed by partial correlations. RESULTS: Our results showed that the plasma NO levels were significantly higher in the diabetic group. When the nondiabetic subjects were analyzed according to their SSPG levels, there was no difference of plasma NO levels between those with SSPG>160 mg/dl and those with SSPG<160 mg/dl. There were also no difference of NO levels between those with a family history of type 2 diabetes and those without. In the nondiabetic group, SSPG correlated with BMI, fasting insulin, triglyceride and HDL cholesterol, but neither with plasma NO levels nor fasting plasma glucose. CONCLUSION: Our data suggests that the impairment of NO activity in patients with type 2 diabetes is due to an impaired effect rather than its production. This altered NO signaling pathway is not an early event in insulin resistant individuals. Any such changes will not be apparent until type 2 diabetes with overt hyperglycemia develops. PMID- 15803116 TI - Changes in plasma triacylglycerol concentrations after sequential lunch and dinner in healthy subjects. AB - OBJECTIVES: The present study examines the kinetic of plasma triacylglycerol (TAG) after sequential ingestion of lunch and dinner as well as the contribution of dietary fat ingested at lunch to subsequent post-dinner TAG composition. METHOD: Six healthy subjects were included. After standardized breakfast (7: 30AM), 2 mixed meals with fat loads composed of 44 g olive oil (rich in oleic acid) at lunch (12PM) and 44 g sunflower oil (rich in linoleic acid) at dinner (7PM) were ingested. [1-13C] palmitate was added in lunch only. Plasma TAG and chylomicron-TAG (CMTAG) levels were measured sequentially after meals. [1-13C] palmitate enrichment and concentrations of oleic acid and linoleic acid were measured in all lipid fractions. RESULT: Post-dinner plasma TAG peak was delayed as compared to lunch (3 hours vs 1 hour, p=0.002) whereas the magnitude of the postprandial peaks was not significantly different between lunch and dinner (2.4+/-0.3 vs 2.0+/-0.4 mmol/L, p=0.85). [1-13C] palmitate enrichment was maximal 5 hours after lunch in all lipid fractions and decreased slowly thereafter. After dinner ingestion, the rate of decline of [1-13C] palmitate enrichment plateaued during the first 60 minutes. Oleic acid increased slightly and immediately after dinner and remained the predominant fatty acid in all lipid fractions during the first hour after dinner. A delayed peak of plasma and CM-TAG was observed after dinner as compared to lunch without difference in the magnitude of peaks. CONCLUSION: The contribution of dietary fat ingested at lunch to post-dinner lipemia is confirmed despite the relatively long lasting interval between the 2 meals (7 h) and the absence of any early peak of plasma TAG after dinner. PMID- 15803117 TI - Scale of therapeutic behavior of type 2 diabetic patients. Hierarchical analysis of a questionnaire. AB - OBJECTIVES: Patient's therapeutic behavior determines the quality of self care in diabetes. The sociological approach can contribute to a better understanding of the internal logic of patient's behavior. The objective of our work is to study patients representations of their illness and its treatment in Moroccan type 2 diabetic patients. Our study concerns 307 type 2 diabetic patients. METHOD: The collection of data has been achieved by means of a questionnaire by investigators with sociological training. The study consists of hierarchical analysis of a questionnaire on representations, attitudes and behavior of patients with the help of Loevinger's coefficient. RESULTS: Test of several groups of hierarchized answer questions allows to identify a 5 questions scale (scale whose coefficient value is 0.47). Otherwise, different social, psychological and therapeutic variables influence the integration of patients in the scale of therapeutic behavior. CONCLUSION: It is possible to recognize a scale of therapeutic behavior in type 2 diabetes. The use of several other sociological qualitative or quantitative methods, by different authors and in different contexts enables to approach the logic of the therapeutic behavior. Its understanding can allows us to adapt the education, the treatment and the follow-up to every patient. PMID- 15803118 TI - Celiac disease in adult patients with type 1 diabetes mellitus in Tunisia. AB - OBJECTIVE: Type1 diabetes mellitus may be associated with celiac disease. The prevalence of celiac disease as determined by screening among adult patients with type 1 diabetes is high with rates of 1.07.8% in Europe and U.S.A. The aims of the study are to determine the prevalence of celiac disease in adults with type 1 diabetes in Tunisia. METHODS: 348 consecutive adult patients with type1 diabetes were investigated prospectively and screened for celiac disease. The mean age was 28.45+/-10.74 years old. There were 176 females and 172 males. For the screening of celiac disease, we used immunoglobulin A (IgA) anti-endomysium (EMA) antibodies determined by an indirect immunofluorescence method. Anti transglutaminase (tTG) antibodies were determined by an ELISA method. Those patients with positive results for anti EMA and or tTG were proposed for duodenal biopsy. RESULTS: 14 patients were positive for anti EMA and had high or a weak positive level of tTG antibodies. One patient from this group was already known to have celiac disease. Only 8 patients consented to biopsy and morphological changes were consistent with celiac disease in all cases. Prevalence of biopsy proven celiac disease was 2.3% (95% CI=1.0-4.5%). CONCLUSION: The present study confirms that celiac disease of adults is prevalent in type 1 diabetic patients in Tunisia. Serological screening for celiac disease in type 1 diabetes is important because many patients are asymptomatic and most are detected by the screening. PMID- 15803119 TI - What could be the reasons for giving up the implanted pump treatment? AB - The development of implanted insulin pump treatment has been particularly intense in France and French implanting centers joined to form an association called EVADIAC. The data collected by EVADIAC have clearly documented benefits as well possible problems with this treatment mode. In comparison with SC administration, intraperitoneal (IP) insulin infusion via an implanted pump achieves good metabolic control, improves long-term stability of diabetes and significantly reduces the risk of severe hypoglycemia. Problems can involve pump slowdown, catheter obstruction or local complication at the implantation site. With respect to the benefits, the rate of complications is highly acceptable. However, amongst the implanted diabetic patients, some elect to give up such a treatment mode. In a retrospective study we intended to examine their reasons for giving up this treatment. In our center, the patients who elected by themselves to give up the implanted pump account for 16% of all treated patients. This group of patients did not have, as compared to other implanted patients, less benefits nor more restrictions or incidents that could have impaired their well being. The reasons for giving up the implanted pump seem more often to be related to the refusal of a strict and institutional management linked to this treatment. PMID- 15803120 TI - Factors related to CSII compliance. AB - Many studies have demonstrated the metabolic efficacy of continuous subcutaneous insulin infusion (CSII) and particularly a reduction of glycaemic fluctuations in type 1 diabetic patients. Despite this benefit, many patients decide to discontinue the use of CSII. To determine the factors related to discontinuation of CSII we analyzed clinical data from a group of 70 patients who had been consecutively started on this treatment from April 2000 to April 2002. Patients were followed for up to 2 years. Eighteen (25.7%) patients decided to terminate CSII during the study after an average of 235 days (range 21-293). The reasons for stopping CSII were decision of the patients (10), end of pregnancy (4), needle site infections (3) and lack of compliance (1). No significant difference was found between patients who had continued and those who had discontinued CSII for age, duration of diabetes, reasons for starting CSII, marital status, prepump concentration of HbA1c and prepump frequency of hypoglycaemia. There tend to be more discontinuations for pregnant women, patients attending hospital visits versus liberal practitioner and patients with lower educational level (below or over baccalaureat) although none of these differences was statistically significant. In conclusion we could not identify any predictive factor of CSII discontinuation. PMID- 15803124 TI - Missed opportunity to ban reproductive cloning. PMID- 15803125 TI - Promoter usage of BRCA1-IRIS. PMID- 15803126 TI - Dual role for Bcl-2 in antibody affinity maturation. PMID- 15803128 TI - The importance of being Smc5/6. PMID- 15803127 TI - Promoter usage of BRCA1-IRIS. PMID- 15803129 TI - DUBing down a tumour suppressor. PMID- 15803130 TI - Rho Rocks PTEN. PMID- 15803131 TI - Cell polarization mechanisms during directed cell migration. PMID- 15803132 TI - Directing NGF's actions: it's a Rap. PMID- 15803134 TI - Active intranuclear movement of herpesvirus capsids. AB - Although small molecules diffuse rapidly through the interphase nucleus, recent reports indicate that nuclear diffusion is limited for particles that are larger than 100 nm in diameter. Given the apparent size limits to nuclear diffusion, there is some debate as to whether the movement of large particles should be attributed to diffusion or to active transport. Here, we show that 125 nm diameter herpes simplex virus 1 (HSV-1) capsids are actively transported within infected nuclei. Movement is directed, temperature- and energy-dependent, sensitive to the putative myosin inhibitor 2,3-butanedione monoxime (BDM) and to actin depolymerization with latrunculin-A, but insensitive to actin depolymerization with cytochalasin-D. PMID- 15803136 TI - IAPs, RINGs and ubiquitylation. AB - The inhibitor of apoptosis (IAP) proteins all contain one or more baculoviral IAP repeat motifs, through which they interact with various other proteins. Many IAPs also have another zinc-binding motif, the RING domain, which can recruit E2 ubiquitin-conjugating enzymes and catalyse the transfer of ubiquitin onto target proteins. The number of targets of IAP-mediated ubiquitylation is increasing and recent results indicate that outcomes following ubiquitylation are tantalizingly complex. As well as regulating other proteins, the IAPs themselves are controlled by ubiquitin-mediated degradation. PMID- 15803137 TI - Communicating with Hedgehogs. AB - Signalling by secreted Hedgehog (Hh) proteins is important for the development of many tissues and organs. Damage to components of the Hh signal-transduction pathway can lead to birth defects and cancer. The Hh proteins are distributed in tissues in a gradient, and cells respond to different thresholds of Hh with distinct responses. The cellular machinery that is responsible for the unique molecular mechanisms of Hh signal transduction has been largely conserved during metazoan evolution. PMID- 15803138 TI - Translational control in stress and apoptosis. AB - Cells respond to stress stimuli through coordinated changes in gene expression. The regulation of translation is often used under these circumstances because it allows immediate and selective changes in protein levels. There are many examples of translational control in response to stress. Here we examine two representative models, the regulation of eukaryotic initiation factor-2alpha by phosphorylation and internal ribosome initiation through the internal ribosome entry site, which illustrate the importance of translational control in the cellular stress response and apoptosis. PMID- 15803139 TI - The cornified envelope: a model of cell death in the skin. AB - The epidermis functions as a barrier against the environment by means of several layers of terminally differentiated, dead keratinocytes - the cornified layer, which forms the endpoint of epidermal differentiation and death. The cornified envelope replaces the plasma membrane of differentiating keratinocytes and consists of keratins that are enclosed within an insoluble amalgam of proteins, which are crosslinked by transglutaminases and surrounded by a lipid envelope. New insights into the molecular mechanisms and the physiological endpoints of cornification are increasing our understanding of the pathological defects of this unique form of programmed cell death, which is associated with barrier malfunctions and ichthyosis. PMID- 15803140 TI - Iron-sulphur cluster biogenesis and mitochondrial iron homeostasis. AB - Iron-sulphur clusters are important cofactors for proteins that are involved in many cellular processes, including electron transport, enzymatic catalysis and regulation. The enzymes that catalyse the formation of iron-sulphur clusters are widely conserved from bacteria to humans. Recent studies in model systems and humans reveal that iron-sulphur proteins have important roles in mitochondrial iron homeostasis and in the pathogenesis of the human disease Friedreich ataxia. PMID- 15803141 TI - Immunotherapy of murine prostate cancer using whole tumor cells killed ex vivo by herpes simplex viral thymidine kinase/ganciclovir suicide gene therapy. AB - Whole cell cancer vaccines are currently under clinical evaluation. Their immunogenicity may depend on the mode of death of the vaccine cells prior to uptake by professional antigen-presenting cells and crosspriming of T cells. Destruction of tumor in vivo by genetic prodrug activation therapy leads to a marked local and systemic immune response, local T-cell infiltration and the establishment of T-cell memory. We postulated that this immunostimulation may be due to induction of danger signals and the inherent immunogenicity of products of HSVtk/ganciclovir kill. Using established models of murine prostate cancer, we have evaluated the efficacy of anti-tumor vaccines comprising irradiated allogeneic or autologous whole cells expressing HSVtK, which are first killed in vitro by prodrug activation using ganciclovir. HSVtk/ganciclovir-induced cell kill was through the induction of apoptosis. The vaccine was found to be effective in both models and superior to traditional irradiated whole tumor cells even after single doses. Protection against tumor challenge was associated with marked proliferative and Th1 cytokine responses. This approach would be applicable clinically in terms of ease of vaccine production, safety, storage and avoidance of potential toxicities of in vivo gene transfer. PMID- 15803142 TI - Effect of EBV LMP1 targeted DNAzymes on cell proliferation and apoptosis. AB - The latent membrane protein (LMP1) encoded by Epstein-Barr virus (EBV) has been suggested to be one of the major oncogenic factors in EBV-mediated carcinogenesis. RNA-cleaving DNA enzymes are catalytic nucleic acids that bind and cleave a target RNA in a highly sequence-specific manner. In this study, we explore the potential of using DNAzymes as a therapeutic approach to EBV associated carcinomas by targeting the LMP1 gene. In all, 13 different phosphorothioate-modified "10-23" deoxyribozymes (DNAzymes) were designed and synthesized against the LMP1 mRNA and transfected into B95-8 cells, which constitutively express the LMP1. Fluorescence microscopy was used to examine the cellular uptake and distribution in B95-8 cells. As demonstrated in Western blots, three out of 13 deoxyribozymes significantly downregulated the expression of LMP1 in B95-8 cells. These DNAzymes were shown to markedly inhibit B95-8 cell growth compared with a disabled DNAzyme and untreated controls, as determined by an alamarBlue Assay. It was further demonstrated that these DNAzymes arrested the B95-8 cells in G0/G1 using flow cytometry. Interestingly, the active DNAzymes could also downregulate the expression of Bcl-2 gene in treated cells, suggesting a close association between the LMP1 and Bcl-2 genes and their involvement in apoptosis. This was further confirmed with the result that the DNAzymes could induce the release of cytochrome c from mitochondria, which is the hallmark of the apoptosis. The present results suggest that the LMP1 may present a potential target for DNAzymes towards the EBV-associated carcinoma through cell proliferation and apoptosis pathways. PMID- 15803143 TI - Intratumoral injection of IL-secreting syngeneic/allogeneic fibroblasts transfected with DNA from breast cancer cells prolongs the survival of mice with intracerebral breast cancer. AB - Prior studies have revealed the immunotherapeutic properties of a vaccine prepared by transfer of genomic DNA from breast cancer cells into a highly immunogenic cell line. The rationale for this type of vaccine is that genes specifying an array of weakly immunogenic, unique tumor antigens associated with the malignant cells will be expressed in a highly immunogenic form by the transfected cells. Here, the immunotherapeutic properties of a vaccine prepared by transfection of mouse fibroblasts with DNA from a breast carcinoma (SB-5b) that arose spontaneously in a C3H/He mouse (H-2Kb) were tested in mice with intracerebral breast cancer. To augment their nonspecific immunogenic properties, before DNA transfer, the fibroblasts (of C3H/He mouse origin) were modified to express allogeneic MHC class I H-2Kb-determinants and to secrete IL-2, IL-18 or GM-CSF. The results indicate that C3H/He mice injected intracerebrally (i.c.) with the breast cancer cells and syngeneic/allogeneic-transfected fibroblasts modified to secrete IL-2 survived significantly longer (P < .005) than mice in various control groups, including mice injected i.c. with the breast cancer cells alone. The immunotherapeutic properties of transfected fibroblasts modified to secrete IL-18 or GM-CSF were less efficacious. The results of two independent in vitro cytotoxicity assays indicate that systemic cellular antitumor immunity was generated in mice injected i.c. with the transfected cells, and the immunity was mediated predominantly by CD8+ T cells. PMID- 15803144 TI - Small interfering RNA targeting Raf-1 inhibits tumor growth in vitro and in vivo. AB - Raf-1 is a cytosolic serine-threonine kinase that plays an important role in tumor cell growth, proliferation, and apoptosis. Upregulated Raf-1 activity has also been implicated in tumor angiogenesis and metastasis. In this study, we used a promising new RNA interfering technology that targets Raf-1 mRNA both in vitro and in vivo. We initially found that Raf-1 siRNA markedly reduced Raf-1 mRNA in MDA-MB-435 cells in vitro by approximately 75% compared to control siRNA treatment groups. Raf-1 siRNA also reduced cell number by inducing apoptosis in a number of cell lines including HUVEC, MDA-MB-435, and C6 cells. After screening several histidine-lysine polymers in complex with Raf-1 siRNA to reduce tumor growth, we further evaluated the efficacy of this siRNA in complex with the optimal histidine-lysine carrier to reduce the tumor growth in vivo. MDA-MB-435 xenografts treated by intratumoral injections of Raf-1 siRNA were significantly reduced compared with the control groups. By the fourth measurement, tumor growth was reduced by nearly 60% in the Raf-1 siRNA treatment group compared with the untreated group (P < .02). Taken together, our data provide evidence that Raf-1 siRNA may be an effective strategy for reducing tumor growth. PMID- 15803145 TI - Optimization of radiation controlled gene expression by adenoviral vectors in vitro. AB - The radiation-inducible EGR-1-promoter has been used in different gene therapy approaches in order to enhance and locally restrict therapeutic efficacy. The aim of this study was to reduce nonspecific gene expression in the absence of irradiation (IR) in an adenoviral vector. Rat rhabdomyosarcoma R1H tumor cells were infected with adenoviral vectors expressing either EGFP or HSV-TK under control of the murine EGR-1 promoter/enhancer. Cells were irradiated at 0-6 Gy. Gene expression was determined by FACS-analysis (EGFP), or crystal violet staining (HSV-TK). The bovine growth hormone polyadenylation signal (BGH pA) was used as insulating sequence and was introduced upstream or upstream and downstream of the expression cassette. Infected R1H cells displayed IR dose dependent EGFP expression. Cells treated with IR, AdEGR.TK and ganciclovir displayed a survival of 17.3% (6 Gy). However, significant gene expression was observed in the absence of IR with EGR.TK and EGR.EGFP constructs. Introduction of BGHpA upstream or upstream and downstream of expression cassette resulted in decreased nonspecific cytotoxicity by a factor of 1.6-2.3 with minor influence on the induced level of cytotoxicity. Introduction of insulating sequences in adenoviral vectors might allow tighter temporospatial control of gene expression by the radiation-inducible EGR-1 promoter. PMID- 15803147 TI - Effects of Onyx-015 among metastatic colorectal cancer patients that have failed prior treatment with 5-FU/leucovorin. AB - Despite recent improvements in the treatment of metastatic colorectal cancer, few patients are cured and the response rates to second-line treatments are poor. Onyx-015, an oncolytic virus, was administered to patients with metastatic colorectal cancer by hepatic artery infusion. No dose-limiting toxicities were observed in the phase I/II studies. Onyx-015 can kill tumor cells by mechanisms that are distinct from chemotherapeutic agents and may therefore have activity among patients who have failed first-line chemotherapy. The 24 patients included in this analysis had failed first-line therapy with 5-FU/leucovorin, 79% of the patients failed two or more regimens and 58% had failed treatment with Irinotecan. Despite the extensive prior therapy, the median survival of these patients was 10.7 months, 46% were alive at 1 year and two patients (8%) had partial responses. In all, 11 patients (46%) had stable disease at the completion of the four planned viral treatments (3 months). The median survival of this group of patients was 19 months, suggesting that stable disease may be an important predictor of benefit with oncolytic viruses. Eight of the 11 patients with stable disease at 3 months demonstrated a unique radiographic pattern of transient enlargement of tumor masses (10-48%) after the initial infusions of Onyx-015, followed by radiographic evidence of extensive tumor necrosis and regression. The initial enlargement and subsequent tumor necrosis resulted in a prolonged time to achieve objective tumor regression. In addition, the transient enlargement of the tumor masses may have resulted in premature removal of responding patients. Treatment of eight patients was stopped prior to completion of the planned four treatments due to presumed progression as defined by standard radiographic criteria (>25% increase in tumor size). Functional imaging, such as positron emission tomography (PET) scans, may help distinguish clinical responses from progressive disease following treatment with oncolytic viruses. Onyx-015 may benefit patients with refractory colorectal cancer and additional studies that include PET scans to assess clinical response are warranted. PMID- 15803146 TI - Epigenetic regulation of protein tyrosine phosphatases: potential molecular targets for cancer therapy. AB - Promoter methylation-mediated silencing is a hallmark of many established tumor suppressor genes. This review focuses on the methylation and suppression of a receptor-type tyrosine phosphatase gene, PTPRO, in a variety of solid and liquid tumors. In addition, PTPRO exhibits many other characteristics of a bona fide tumor suppressor. Reactivation of genes silenced by methylation using inhibitors of DNA methyltransferases and histone deacetylases, and the potential application of PTPRO as a molecular target for cancer therapy have been discussed. PMID- 15803148 TI - TEC-family kinases: regulators of T-helper-cell differentiation. AB - The TEC-family protein tyrosine kinases ITK, RLK and TEC have been identified as key components of T-cell-receptor signalling that contribute to the regulation of phospholipase C-gamma, the mobilization of Ca(2+) and the activation of mitogen activated protein kinases. Recent data also show that TEC kinases contribute to T cell-receptor-driven actin reorganization and cell polarization, which are required for productive T-cell activation. Functional studies have implicated TEC kinases as important mediators of pathways that control the differentiation of CD4(+) T helper cells. Here, we review studies of signalling pathways that involve TEC kinases and how these pathways might contribute to the regulation of T-helper-cell differentiation and function. PMID- 15803149 TI - Dendritic cells as therapeutic vaccines against cancer. AB - Mouse studies have shown that the immune system can reject tumours, and the identification of tumour antigens that can be recognized by human T cells has facilitated the development of immunotherapy protocols. Vaccines against cancer aim to induce tumour-specific effector T cells that can reduce the tumour mass, as well as tumour-specific memory T cells that can control tumour relapse. Owing to their capacity to regulate T-cell immunity, dendritic cells are increasingly used as adjuvants for vaccination, and the immunogenicity of antigens delivered by dendritic cells has now been shown in patients with cancer. A better understanding of how dendritic cells regulate immune responses will allow us to better exploit these cells to induce effective antitumour immunity. PMID- 15803150 TI - The zebrafish: a new model of T-cell and thymic development. AB - T-cell and thymic development are processes that have been highly conserved throughout vertebrate evolution. Mammals, birds, reptiles and fish share common molecular signalling pathways that regulate the development of the adaptive immune system. This Review article focuses on defining the similarities and differences between zebrafish and mammalian T-cell immunobiology, and it highlights the advantages of using the zebrafish as a genetic model to uncover mutations that affect T-cell and thymic development. Finally, we summarize the use of the zebrafish as a new model for assessing stem-cell function and for drug discovery. PMID- 15803151 TI - Heat-shock proteins induce T-cell regulation of chronic inflammation. AB - Immune responses to certain heat-shock proteins (HSPs) develop in almost all inflammatory diseases; however, the significance of such responses is only now becoming clear. In experimental disease models, HSPs can prevent or arrest inflammatory damage, and in initial clinical trials in patients with chronic inflammatory disease, HSP-derived peptides have been shown to promote the production of anti-inflammatory cytokines, indicating that HSPs have immunoregulatory potential. In this Review, we discuss the unique characteristics of HSPs that endow them with these immunoregulatory qualities. PMID- 15803152 TI - High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. AB - High-mobility group box 1 protein (HMGB1), which previously was thought to function only as a nuclear factor that enhances transcription, was recently discovered to be a crucial cytokine that mediates the response to infection, injury and inflammation. These observations have led to the emergence of a new field in immunology that is focused on understanding the mechanisms of HMGB1 release, its biological activities and its pathological effects in sepsis, arthritis, cancer and other diseases. Here, we discuss these features of HMGB1 and summarize recent advances that have led to the preclinical development of therapeutics that modulate HMGB1 release and activity. PMID- 15803153 TI - Mechanisms of B-cell lymphoma pathogenesis. AB - Chromosomal translocations involving the immunoglobulin loci are a hallmark of many types of B-cell lymphoma. Other factors, however, also have important roles in the pathogenesis of B-cell malignancies. Most B-cell lymphomas depend on the expression of a B-cell receptor (BCR) for survival, and in several B-cell malignancies antigen activation of lymphoma cells through BCR signalling seems to be an important factor for lymphoma pathogenesis. Recent insights into the lymphomagenic role of factors supplied by the microenvironment also offer new therapeutic strategies. PMID- 15803154 TI - Tumour stem cells and drug resistance. AB - The contribution of tumorigenic stem cells to haematopoietic cancers has been established for some time, and cells possessing stem-cell properties have been described in several solid tumours. Although chemotherapy kills most cells in a tumour, it is believed to leave tumour stem cells behind, which might be an important mechanism of resistance. For example, the ATP-binding cassette (ABC) drug transporters have been shown to protect cancer stem cells from chemotherapeutic agents. Gaining a better insight into the mechanisms of stem cell resistance to chemotherapy might therefore lead to new therapeutic targets and better anticancer strategies. PMID- 15803155 TI - Chemical approaches to the discovery and development of cancer therapies. AB - The chemical sciences are essential for the process of anticancer-drug discovery, and a range of chemical research techniques is needed to develop clinically effective drugs. Improved understanding of the cellular, molecular and genetic basis of cancer has increased the number of drug targets available. What chemical approaches are used to develop agents that target specific features of cancer cells and make these therapeutics more effective? We outline the roles that chemical synthesis and understanding of drug uptake have had in drug discovery over the past 100 years, as well as the chemical insights derived from knowledge of the three-dimensional structure of targets. PMID- 15803156 TI - Nuclear factor-kappaB inhibitors as sensitizers to anticancer drugs. AB - The cytotoxicity of chemotherapeutic agents is attributed to apoptosis. Acquired resistance to the effects of chemotherapy has emerged as a significant impediment to effective cancer therapy. One feature that cytotoxic treatments of cancer have in common is their activation of the transcription factor nuclear factor-kappaB (NF-kappaB), which regulates cell survival. NF-kappaB activation suppresses the apoptotic potential of chemotherapeutic agents and contributes to resistance. What evidence is there that inhibitors of NF-kappaB might promote apoptosis in cancer cells and can NF-kappaB inhibitors be used to overcome resistance to chemotherapeutic agents? PMID- 15803157 TI - Leukaemia stem cells and the evolution of cancer-stem-cell research. AB - Many cancers seem to depend on a small population of 'cancer stem cells' for their continued growth and propagation. The leukaemia stem cell (LSC) was the first such cell to be described. The origins of these cells are controversial, and their biology - like that of their normal-tissue counterpart, the haematopoietic stem cell (HSC) - is still not fully elucidated. However, the LSC is likely to be the most crucial target in the treatment of leukaemias, and a thorough understanding of its biology - particularly of how the LSC differs from the HSC - might allow it to be selectively targeted, improving therapeutic outcome. PMID- 15803158 TI - The role of calmodulin as a signal integrator for synaptic plasticity. AB - Excitatory synapses in the brain show several forms of synaptic plasticity, including long-term potentiation (LTP) and long-term depression (LTD), which are initiated by increases in intracellular Ca(2+) that are generated through NMDA (N methyl-D-aspartate) receptors or voltage-sensitive Ca(2+) channels. LTP depends on the coordinated regulation of an ensemble of enzymes, including Ca(2+)/calmodulin-dependent protein kinase II, adenylyl cyclase 1 and 8, and calcineurin, all of which are stimulated by calmodulin, a Ca(2+)-binding protein. In this review, we discuss the hypothesis that calmodulin is a central integrator of synaptic plasticity and that its unique regulatory properties allow the integration of several forms of signal transduction that are required for LTP and LTD. PMID- 15803159 TI - Dendritic spines and long-term plasticity. AB - A recent flurry of time-lapse imaging studies of live neurons have tried to address the century-old question: what morphological changes in dendritic spines can be related to long-term memory? Changes that have been proposed to relate to memory include the formation of new spines, the enlargement of spine heads and the pruning of spines. These observations also relate to a more general question of how stable dendritic spines are. The objective of this review is to critically assess the new data and to propose much needed criteria that relate spines to memory, thereby allowing progress in understanding the morphological basis of memory. PMID- 15803160 TI - Normal and pathological oscillatory communication in the brain. AB - The huge number of neurons in the human brain are connected to form functionally specialized assemblies. The brain's amazing processing capabilities rest on local communication within and long-range communication between these assemblies. Even simple sensory, motor and cognitive tasks depend on the precise coordination of many brain areas. Recent improvements in the methods of studying long-range communication have allowed us to address several important questions. What are the common mechanisms that govern local and long-range communication and how do they relate to the structure of the brain? How does oscillatory synchronization subserve neural communication? And what are the consequences of abnormal synchronization? PMID- 15803161 TI - Anatomical and physiological foundations of cerebellar information processing. AB - A coordinated movement is easy to recognize, but we know little about how it is achieved. In search of the neural basis of coordination, we present a model of spinocerebellar interactions in which the structure-functional organizing principle is a division of the cerebellum into discrete microcomplexes. Each microcomplex is the recipient of a specific motor error signal - that is, a signal that conveys information about an inappropriate movement. These signals are encoded by spinal reflex circuits and conveyed to the cerebellar cortex through climbing fibre afferents. This organization reveals salient features of cerebellar information processing, but also highlights the importance of systems level analysis for a fuller understanding of the neural mechanisms that underlie behaviour. PMID- 15803162 TI - Cortical inhibitory neurons and schizophrenia. AB - Impairments in certain cognitive functions, such as working memory, are core features of schizophrenia. Convergent findings indicate that a deficiency in signalling through the TrkB neurotrophin receptor leads to reduced GABA (gamma aminobutyric acid) synthesis in the parvalbumin-containing subpopulation of inhibitory GABA neurons in the dorsolateral prefrontal cortex of individuals with schizophrenia. Despite both pre- and postsynaptic compensatory responses, the resulting alteration in perisomatic inhibition of pyramidal neurons contributes to a diminished capacity for the gamma-frequency synchronized neuronal activity that is required for working memory function. These findings reveal specific targets for therapeutic interventions to improve cognitive function in individuals with schizophrenia. PMID- 15803163 TI - Statins--a cure-all for the brain? AB - 'Statins' are 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors - oral cholesterol-lowering drugs that are used to treat hypercholesterolaemia. It is widely accepted that statins have anti-inflammatory effects that are independent of their ability to lower cholesterol. Animal studies and observational clinical studies have indicated that statins might also be effective in treating certain neurological diseases - in particular, multiple sclerosis, Alzheimer's disease and ischaemic stroke. At present, however, results from ongoing prospective, randomized clinical trials are not available. PMID- 15803164 TI - From presence to consciousness through virtual reality. AB - Immersive virtual environments can break the deep, everyday connection between where our senses tell us we are and where we are actually located and whom we are with. The concept of 'presence' refers to the phenomenon of behaving and feeling as if we are in the virtual world created by computer displays. In this article, we argue that presence is worthy of study by neuroscientists, and that it might aid the study of perception and consciousness. PMID- 15803165 TI - In vivo confocal microscopy in congenital rubella keratopathy. PMID- 15803166 TI - Long-term intraocular pressure control of eyes that developed encapsulated blebs following trabeculectomy. AB - PURPOSE: To evaluate the long-term intraocular pressure (IOP) control of eyes that developed an encapsulated bleb (EB) following trabeculectomy. METHODS: Between 1994 and 1995, 25 eyes developed EBs and were randomized to medical treatment or needling without adjunct antimetabolites. Among the 25 patients who developed an EB, 21 were followed for at least 6 months. A control group of 21 consecutive eyes, which underwent trabeculectomy during the same period and that did not develop EBs was retrospectively selected. Success was defined as IOP <21 mmHg with or without medications. Kaplan-Meier survival analysis was performed to compare the groups. RESULTS: Among the 21 eyes that developed EBs, 12 (57%) had undergone transconjunctival needling and nine (43%) had received medical treatment. Mean follow-ups were 30.0 +/- 14.0 months, 33.3 +/- 18.5 months, and 37.4 +/- 2.6 months for the needling, medical treatment, and control groups, respectively (P = 0.19). Kaplan-Meier survival curves demonstrated that the control group showed a significantly lower chance of failure than both the needling and the medical treatment groups (P < 0.0001). CONCLUSION: Encapsulated blebs may be associated with an increased risk for surgical failure. PMID- 15803167 TI - Therapeutic corneal tattoo following peripheral iridotomy complication. PMID- 15803168 TI - Conjunctival ischaemia subsequent to posterior subtenon's triamcinolone acetonide injection. PMID- 15803169 TI - Resting pulse rates in a glaucoma clinic: the effect of topical and systemic beta blocker usage. AB - INTRODUCTION: Beta-blockers have, for 25 years, been a commonly used agent in the treatment of raised intraocular pressure (IOP). Beta-blockers can also reduce the pulse rate. With no available literature examining a cohort of patients, this study aims to investigate the resting pulse rates in patients attending a specialist glaucoma clinic in order to identify if routine review of ophthalmic medication use is indicated. METHOD: The resting pulse rates of patients attending a glaucoma clinic were measured using pulseoximetry, with a medical and drug history established for each patient. RESULTS: In all, 205 patients were included in the study. A total of 101 (49%) of patients were using beta-blockers in some form. The mean pulse rate for patients not using beta-blockers (104 patients) was 76 beats per minute (bpm), for topical use only (68 patients) it was 70.3 bpm, for oral use (18 patients) it was 64.7 bpm, and 58 bpm for patients using both topical and oral beta-blockers (15 patients). Groups using beta blockers (oral, topical, oral and topical) were considered in relation to patients not using beta-blockers. All groups using beta-blockers showed a significant association with causing a bradycardia of less than 60 bpm. Patients with a pulse rate of less than 50 bpm were significantly more likely to be using topical and oral beta-blockers than oral beta-blockers alone (P=0.01). CONCLUSION: Topical beta-blockers should be used with caution, even in the presence of established systemic beta-blocker use. Routine pulse rate monitoring and review of ophthalmic medication are indicated in patients using beta-blocker therapy. PMID- 15803170 TI - Optic nerve axons: life and death before birth. AB - In the 2004 Bowman Lecture, I give a panegyric for Sir William Bowman, an estimate of the importance and the epidemiology of anterior visual pathway developmental disorders, followed by a history of the anterior visual system. I review the normal development of the optic nerve and chiasm and the main developmental disorders: Optic Nerve Aplasia, Optic Nerve Hypoplasia and Achiasmia. PMID- 15803171 TI - A rationale for membrane peeling in the repair of stage 4 macular holes. AB - AIM: To examine the histological and immunocytochemical characteristics of epiretinal membranes (ERM) associated with stage 4 macular holes (MH) so as to establish a vitreoretinal rationale for surgery in stage 4 MH. METHOD: Consecutive patients with stage 4 MH undergoing vitrectomy and membrane peeling were recruited. Preoperatively, the eyes were examined for ERM formation over the macula and completeness of posterior hyaloid membrane (PHM) separation from the retina. ERM peel specimens obtained during surgery were sent for histological and immunocytochemical studies and were compared with the PHM specimens taken from a previous post-mortem study of eyes with physiological posterior vitreous detachment but without macular holes. RESULT: A total of 13 patients with stage 4 MH fulfilled the inclusion criteria and were recruited. Preoperatively, all eyes had an ERM over the macula and incomplete separation of the PHM seen as a defect in the PHM on specular biomicroscopy. Histologically, the ERM specimens had very similar morphological characteristics to PHM, consisting of an eosinophilic membrane of varying thickness with scattered spindle-shaped cells. The membranes stained positively for type IV collagen while the cells were glial fibrillary acidic protein positive. Postoperatively, successful closure of MH was achieved in all cases. CONCLUSION: Stage 4 MH is characterised by incomplete separation of the PHM from the retina with remnants overlying the macula manifesting as ERM. Removal of the ERM is required during vitrectomy in order to relieve the tangential forces involved in the development of MH. PMID- 15803172 TI - Plasma homocysteine and total thiol content in patients with exudative age related macular degeneration. AB - PURPOSE: Exudative age-related macular degeneration (ARMD) is one of the debilitating ocular complications, which results in permanent blindness. Elevated homocysteine (Hcys) levels have been associated in the development of several vascular diseases. Vascular and oxidative stress theories have been implicated for the development of choroidal neovascularization in exudative ARMD. The aim of the present study was to investigate the possible role of plasma Hcys and thiol content (tSH) as a risk factor for the development of exudative ARMD. METHOD: A total of 16 patients with exudative ARMD and 20 age-matched controls were recruited for the study. Plasma Hcys levels were analysed using Reverse Phase High Performance Liquid Chromatography. Plasma glutathione (GSH) content was determined using o-phthalaldehyde (OPA) derivatization and subsequent detection by fluorimeter. Plasma tSH levels were determined by using thiol-specific reagent dithionitrobenzoic acid (DTNB) spectrophotometrically. RESULTS: Plasma Hcys levels in exudative ARMD were elevated three-fold (18+/-5.0 microM) when compared to healthy controls (6.7+/-1.8 microM). There was a two-fold decrease in the GSH and tSH in exudative ARMD when compared with controls. Negative correlation was observed between diminished tSH and Hcys levels (r=-0.4837, P=0.05). Similarly plasma Hcys levels negatively correlated with GSH content (r=-0.6620, P<0.05). CONCLUSION: Results from our present study revealed that there is an elevated Hcys level and diminished thiol pool content in exudative ARMD that are significant. PMID- 15803173 TI - Comparison of intraocular pressure measured by Pascal dynamic contour tonometry and Goldmann applanation tonometry. AB - AIMS: To compare the intraocular pressure (IOP) measurements obtained using the Pascal dynamic contour tonometer (PDCT) with the standard Goldmann applanation tonometer (GAT) and to correlate these with central corneal thickness (CCT) in patients with normal corneas. METHODS: A prospective, masked, comparative case series of 116 eyes from patients attending a glaucoma clinic. IOP was measured with PDCT by one examiner and with GAT by a masked, independent examiner. A mean of six CCT readings was used for analysis. RESULTS: IOP measured by the two instruments correlated significantly (r=0.77; P<0.0001). IOP measured by GAT correlated strongly with CCT (r=0.37, P=0.0001) whereas the relationship between IOP measured by PDCT and CCT approached significance (r=0.17, P=0.073). The differences between GAT and PDCT measured IOP also correlated strongly with CCT (r=0.37, P<0.0001). The 95% limits of agreement between GAT and PDCT were +/-4.2 mmHg. Dividing the eyes into three groups on the basis of CCT, demonstrated those in the thickest tertile showed a poorer agreement between instruments and the GAT measured significantly higher IOP in this group (P=0.003) while the PDCT showed no significant differences with different CCTs (P=0.37). CONCLUSION: Demonstration of the relative independence of PDCT IOP measurements from CCT supports a potential clinical role for this instrument, particularly for subjects with CCT outside the normal range. PMID- 15803174 TI - Primary peripapillary membrane in an 8-year-old boy. PMID- 15803175 TI - Vitrectomy for juvenile uveitis: prognostic factors for the long-term functional outcome. AB - PURPOSE: This study evaluated the long-term effect of pars plana vitrectomy (PPV) in children and adolescents with chronic uveitis on visual function, anatomical outcome, and the requirement of systemic treatment. Further, predictive preoperative factors associated with a beneficial visual outcome were assessed. METHODS: Retrospective review of 29 eyes of 23 consecutive paediatric and juvenile patients below 20 years of age with chronic uveitis who underwent a PPV for visually significant opacities in 25 eyes, vitreous haemorrhage in three eyes, and retinal detachment in one eye. The clinical diagnosis was chronic intermediate uveitis in 22 eyes and retinal vasculitis of different origin in seven eyes. RESULTS: LogMAR visual acuity improved from an average of 0.91 to 0.33 (P<0.001). Cystoid macular oedema (CME) was significantly reduced in eight of 10 eyes postoperatively (P=0.021). In the multiple regression analysis, a low preoperative logMAR visual acuity and the presence of a CME had a negative influence on the final logMAR visual acuity. Furthermore, the appearance of chronic uveitis relapses was significantly reduced from 15 eyes before to seven eyes after surgery (P=0.042). CONCLUSIONS: PPV has a beneficial effect on the course and the complications of chronic uveitis in paediatric and juvenile patients with respect to the anatomical and visual outcome. Preoperative logMAR visual acuity and clinically significant CME were the most accurate predictors for the functional outcome. PMID- 15803176 TI - ROP was always there. PMID- 15803177 TI - Purtscher's like retinopathy as the presenting feature of acute alcoholic pancreatitis. PMID- 15803178 TI - Superior forniceal conjunctival advancement pedicles (SFCAP) in the management of acute and impending corneal perforations. AB - OBJECTIVE/AIM: Corneal perforations can result from a wide variety of disorders and can lead to devastating visual sequelae. Various surgical procedures have been described to manage nontraumatic corneal perforation. Conjunctival flaps offer an important technique in dealing with such corneal emergencies. We report a modified conjunctival flap procedure referred to as superior forniceal conjunctival advancement pedicle (SFCAP) in the successful management of corneal perforation and impending corneal perforation. PATIENTS AND METHODS: Out of 20 patients who underwent SFCAP for non-traumatic corneal perforation and impending perforation, 16 had corneal perforations. Perforations were secondary to acne rosacea (2), advanced bacterial keratitis (6), corneal anaesthesia(2), multiple retinal procedures (2), previous corneal grafts with a compromised ocular surface (3), and advanced Mooren's ulcer (1). RESULTS: The globe was preserved in all patients. In 14 of the 16 eyes with perforated corneas the pedicle stabilised. One patient, who was a chronic alcoholic, rubbed the eye during sleep detaching the pedicle a week after surgery. However, the ulcer healed and the integrity of the globe was restored. None of these patients developed secondary glaucoma or ptosis. CONCLUSION: SFCAP is an appropriate procedure to manage corneal perforations and impending corneal perforations where donor material is not available and transplantation of such tissue is not suitable. PMID- 15803179 TI - Retinal pigment epithelial tear following photodynamic therapy for choroidal neovascularization secondary to AMD. AB - PURPOSE: To describe retinal pigment epithelial tear following photodynamic therapy (PDT) for subfoveal choroidal neovascularization (CNV) secondary to age related macular degeneration (AMD). DESIGN: Retrospective interventional case series. METHODS: A retrospective study in an institutional practice. We describe seven cases of retinal pigment epithelial (RPE) tear, which developed in seven eyes of seven patients following PDT. All eyes had subfoveal CNV secondary to AMD. RESULTS: Six eyes had occult subfoveal CNV, and one eye had recurrent classic subfoveal CNV. In five patients, the eye that developed the tear was the second eye, whereas the first eye had a disciform scar. In four eyes, the RPE tear developed after one PDT, in one eye the RPE tear developed after the second PDT, and in two eyes the RPE tear developed after the third PDT. In five of seven cases, there was a significant visual deterioration following the RPE tear. CONCLUSIONS: RPE tear is a complication that may occur following PDT in particular when the PDT is applied to an occult subfoveal CNV. PMID- 15803180 TI - Retinoblastoma-binding protein 2-homolog 1: a retinoblastoma-binding protein downregulated in malignant melanomas. AB - In malignant melanomas, the loss of cell cycle control is thought to be due to a lack of retinoblastoma protein (pRb)-activity. Members of the previously described family of retinoblastoma-binding proteins (RBPs) are supposed to act as pRb-modulating factors. Based on RNA-fingerprinting of normal human melanocytes, we previously described a new family member with high sequence homology to the retinoblastoma-binding protein-2 (RBP-2), termed RBP2-Homolog 1 (RBP2-H1). Based on its UVB responsiveness, it was hypothesized that this gene may also play a role in melanocytic tumors. In the present study, we can confirm by real-time RT PCR (six common melanocytic nevi, five advanced nodular melanomas and seven melanoma metastases) and immunohistochemistry (tissue microarrays: 52 melanocytic nevi, 60 melanomas, 60 metastases; and conventional sections: five common nevi, four advanced nodular melanomas, five melanoma metastases) that RBP2-H1 expression is progressively downregulated in advanced and metastatic melanomas in vivo with a certain intratumoral heterogeneity. Whereas benign melanocytic nevi are RBP2-H1 positive in about 70% of the cases, a lack of RBP2-H1 expression was found in 90% of the primary malignant melanomas and 70% of the melanoma metastases, respectively. Interestingly, a similar deficiency can be found in glioblastomas, but not epithelial cancers. In accordance to the in vivo data, established melanoma cell lines exhibit low but heterogeneous levels of RBP2-H1 expression. By co-immunoprecipitation, we provide the first evidence that a subfraction of total RBP2-H1 can bind to pRb, which makes this protein a true pRb interacting factor. We conclude that loss of RBP2-H1 is a common finding in the progression of malignant melanomas. Since a direct interaction of RBP2-H1 and pRb seems possible, the loss of RBP2-H1 may possibly contribute to uncontrolled growth in malignant melanomas. PMID- 15803181 TI - Eosinophilic cell change of the endometrium: a possible relationship to mucinous differentiation. AB - Eosinophilic cell change is one of the most common endometrial metaplasias occurring in both non-neoplastic and neoplastic endometrium. Its phenotypic characteristics have not still been fully clarified. We examined expression of mucin core proteins in a total of 95 distinct histological areas of endometrial specimens comprising 39 benign nonhyperplastic endometria, 14 endometrial hyperplasias, and 42 endometrial carcinomas. Eosinophilic cell change was very common, seen in 27 endometrial areas (28%); mucinous metaplasia (28%) and ciliated (tubal) change (31%), were also frequently seen. Eosinophilic cell change was more frequently seen in endometrial hyperplasia and carcinoma than in benign nonhyperplastic endometrium. In endometrial carcinomas, eosinophilic cell change was frequently associated with mucinous metaplasia and the two types of metaplastic cells were occasionally intermingled in a single neoplastic gland. A total of 23 (85%) of 27 eosinophilic cell changes and 18 (72%) of 25 mucinous metaplasias showed MUC5AC expression. These frequencies of MUC5AC expression did not differ significantly among benign non-hyperplastic endometrium, endometrial hyperplasia and endometrial carcinoma. Totally, 15 (52%) of 29 ciliated (tubal) changes and two (100%) of two surface syncytial changes, which showed cytoplasmic eosinophilia at least focally, also expressed MUC5AC. Most of the endometrial changes characterized by cytoplasmic eosinophilia may be subtypes of immature mucinous metaplasia which express a mucin core protein but are not fully glycosylated. PMID- 15803182 TI - Tumor lymphangiogenesis predicts melanoma metastasis to sentinel lymph nodes. AB - Cutaneous melanoma is a common melanocytic neoplasm that can quickly metastasize to regional lymph nodes. Currently, prognosis is determined by measuring tumor thickness but more reliable markers for metastatic spread are urgently needed. We investigated whether the extent of tumor lymphangiogenesis can predict melanoma metastasis to sentinel lymph nodes. We quantified the extent of tumor lymphangiogenesis, as well as other factors, in excised primary tumors and in sentinel lymph node biopsy samples from 45 patients with primary cutaneous melanoma. The results were correlated with histological and clinical outcome. Primary melanomas from patients whose tumors had metastasized to the sentinel lymph nodes contained prominent 'hot spots' of increased lymphatic vessel density, compared to nonmetastatic tumors. Multivariate risk analysis revealed that the lymphatic vascular area of primary melanomas, an index of tumor lymphangiogenesis, was the most sensitive prognostic marker for sentinel lymph node metastasis, and was even able to more accurately predict which tumors were metastatic to sentinel lymph nodes than the currently used method of measuring tumor thickness. Highly lymphangiogenic melanomas maintained their lymphangiogenic activity after metastasis to the sentinel lymph node. The extent of tumor lymphangiogenesis is a highly sensitive (83%) and specific (89%) prognostic marker of lymph node metastasis. Assessment of lymphangiogenesis in primary melanomas may be a more effective approach than the currently used technique of measuring tumor thickness in selecting patients with early metastatic disease for aggressive therapy. PMID- 15803183 TI - Predominance of high-grade pathway in breast cancer development of Middle East women. AB - Recent data have suggested considerable molecular differences in cancers from various ethnical groups. As molecular features are increasingly used for predicting cancer prognosis and response to therapy, better knowledge of ethnic molecular features is important. To identify potential molecular differences between breast cancers in Europe and the Middle East, we analyzed consecutive breast cancer series from Switzerland (n=2197) and Saudi Arabia (n=204). Tissue microarrays were analyzed by fluorescence in situ hybridization for HER2, CCND1, MYC, and EGFR amplification. The data revealed marked differences between Saudi and Swiss patients. Saudi breast cancers had a markedly higher frequency of HER2 (31 vs 17%; P<0.0001) and MYC (16 vs 5%; P<0.0001) amplifications than Swiss breast cancers. Remarkably, this was partly due to a much higher incidence of grade 3 cancers in the Saudi than in the Swiss population (65 vs 32%; P<0.0001). However, differences in amplification frequency hold also true within grade 3 cancers (HER2: 40 vs 30%, P<0.05; MYC: 22 vs 11%, P=0.002). Interestingly, in combination with known age standardized incidence rates of breast cancer in Saudi Arabia (21.6/100 000) and Switzerland (70.1/100 000), these data suggest that the incidence of high-grade breast cancer is comparable for Saudi and Swiss women, while the incidence of low-grade breast cancers is about 14 times lower in Saudi than for Swiss women. These observations suggest that a difference in genetic susceptibility and/or lifestyle between Saudi and Swiss women has a substantial and much higher than expected impact on the risk of low-grade breast cancer. PMID- 15803184 TI - Value of CDX2, villin, and alpha-methylacyl coenzyme A racemase immunostains in the distinction between primary adenocarcinoma of the bladder and secondary colorectal adenocarcinoma. AB - Primary adenocarcinoma of the urinary bladder is an uncommon neoplasm that can be indistinguishable morphologically from colorectal adenocarcinoma secondarily involving the bladder by direct extension or metastasis. In the current study, 17 enteric-type primary adenocarcinomas of the bladder were immunohistochemically examined for the expression of CDX2, villin and alpha-methylacyl coenzyme A racemase (AMACR), immunomarkers preferentially expressed in colorectal adenocarcinoma. For comparison, 17 secondary colorectal adenocarcinomas involving the bladder, 23 primary colorectal adenocarcinomas and 14 conventional urothelial carcinomas were similarly studied. The results show that all 40 (100%) colorectal adenocarcinomas expressed CDX2 and 39 (98%) expressed villin. The expression of these two immunomarkers was less frequent in primary bladder adenocarcinomas, observed in eight (47%) and 11 (65%) cases, respectively (P<0.0001 and P=0.0019, respectively). The frequency of positive AMACR immunostaining was similar between these two types of tumors, detected in 28 (70%) colorectal adenocarcinomas and 11 (65%) primary bladder adenocarcinomas (P=0.694). None of the urothelial carcinomas exhibited CDX2 or villin immunoreactivity; and only two (14%) showed positive staining for AMACR. These results demonstrate that CDX2 and villin are of diagnostic value in aiding in the distinction between primary adenocarcinoma of the bladder and secondary colorectal carcinoma. Lack of CDX2 and villin signals points strongly to a bladder primary. PMID- 15803185 TI - Cellular hamartoma resembling gastrointestinal stromal tumor: a solid tumor of the pancreas expressing c-kit (CD117). AB - Solid tumors of the pancreas are usually neoplastic. We report on two adult patients, each with a solid tumor of the pancreas that presented with an unusual histology and seemed to follow a benign course. The tumors, one located in the body and one in the tail, were well demarcated and composed of irregularly arranged but well-differentiated acini and small intralobular and interlobular ducts embedded in predominantly hypocellular fibrotic tissue that contained fascicles of cytologically bland spindle cells. Islets were lacking, but immunohistochemical staining for chromogranin A and insulin revealed individual scattered insulin-producing cells distributed between acinar and ductal cells. The spindle cell component tissue showed coexpression of CD34, c-kit (CD117) and bcl-2. The follow-up (2 and 4 years) of the patients was uneventful. We propose to designate the tumors as 'cellular hamartoma resembling gastrointestinal stromal tumor. PMID- 15803186 TI - Maximum tumor diameter is an independent predictor of prostate-specific antigen recurrence in prostate cancer. AB - Maximum tumor diameter has been shown to correlate with multiple predictors of clinical outcome in prostate cancer. In the current study, we prospectively analyze whether maximum tumor diameter is a significant predictor of prostate specific antigen (PSA) recurrence. The study population consisted of 364 patients who underwent radical prostatectomy for prostate cancer. Prostatectomy specimens were evaluated by whole-mount processing of the entire prostate. Maximum tumor diameter was measured from the whole-mount sections of the prostate. Spearman's coefficient of rank correlation was used to correlate tumor diameter with continuous variables. T-tests or analysis of variance (ANOVA) tests were performed to determine if tumor diameter was significantly associated with other clinical and pathologic variables. The effect of clinical and pathologic variables on time to recurrence was analyzed using Cox regression. The mean tumor diameter for all patients was 1.73 cm (range, 0.02-4.40 cm). Maximum tumor diameter was associated with preoperative PSA (r=0.22, P<0.0001), prostate weight (r=-0.12, P=0.028), tumor volume (r=0.87, P<0.0001), Gleason score (r=0.29, P<0.0001), and pathologic stage (P<0.0001). Cox multiple regression was performed to test the prognostic value of maximum tumor diameter adjusting for pathologic stage, Gleason score, and surgical margin status. Increased maximum tumor diameter was associated with shorter time to PSA recurrence (hazard ratio=1.70, 95% confidence interval 1.13-2.56, P=0.01), controlling for risk factors, Gleason score, and surgical margin status. We conclude that maximum tumor diameter is a significant predictor of biochemical recurrence in patients with prostate cancer. PMID- 15803187 TI - Duodenal intraepithelial lymphocytosis with normal villous architecture: common occurrence in H. pylori gastritis. AB - We have observed expansions of intraepithelial lymphocytes in duodenal biopsies from patients with Helicobacter pylori gastritis. This study was undertaken to prospectively evaluate, unselected, paired gastric and duodenal biopsies from 50 patients with H. pylori gastritis and a comparison group of 30 patients with other types of gastritis (10 autoimmune and 20 reactive) to: (1) quantify duodenal intraepithelial lymphocytes, determine their distribution patterns, epithelial location, and phenotype, and (2) correlate the intraepithelial lymphocyte elevations with various features of gastric and duodenal pathology. Intraepithelial lymphocytes were analyzed with antibodies including CD3, CD8, and TIA-1. A stain for H. pylori was performed on all gastric and duodenal biopsies. Duodenal intraepithelial lymphocytes from patients with H. pylori gastritis (using CD3) ranged from 3 to 42 lymphocytes/100 epithelial cells (mean 18.5) compared to 3 to 18 lymphocytes/100 epithelial cells (mean 6.6) in the comparison group. Intraepithelial lymphocyte elevations were seen in 44% of the duodenal biopsies from patients with H. pylori gastritis (using CD3). Significant differences in the intraepithelial lymphocyte counts between patients with H. pylori gastritis and the comparison group were seen for all three T-cell antigens (P<0.001 for CD3 and CD8 and P<0.002 for TIA-1). Duodenal intraepithelial lymphocytes in the H. pylori+ cases had a latent cytotoxic phenotype, H. pylori was not visualized in any of the duodenal biopsies from patients with H. pylori gastritis, and no patient had clinical evidence of celiac disease. Our study highlights frequent duodenal intraepithelial lymphocytosis in individuals with H. pylori gastritis and the lymphocyte distribution patterns (and numbers) overlapped with those described for celiac disease patients. H. pylori gastritis must be considered as a possible explanation for duodenal intraepithelial lymphocytosis with normal villous architecture, especially when lymphocytosis is patchy, intraepithelial lymphocytes display a 'latent' cytotoxic phenotype, and the clinical findings and serologic profile does not fit celiac disease. PMID- 15803188 TI - VEGF-D expression and lymph vessels play an important role for lymph node metastasis in papillary thyroid carcinoma. AB - Papillary thyroid carcinoma frequently metastasizes to regional lymph nodes, and lymph node metastasis increases the risk of local regional relapse. Recent evidence suggests that vascular endothelial growth factor-D (VEGF-D) promotes lymphangiogenesis, which in turn promotes lymphatic metastasis. Therefore, the role of VEGF-D messenger RNA transcript levels and VEGF-D immunoreactivity in lymph node metastasis in papillary thyroid carcinoma was investigated. In addition, the role of blood vascular vessel, lymph vessel, and Flt-4-positive vessel densities were studied in relation to their suspected association with lymph node metastasis, and with VEGF-D expression. VEGF-D messenger RNA transcript levels by quantitative real-time reverse transcription-polymerase chain reaction and VEGF-D immunoreactivity by immunohistochemistry in 49 papillary thyroid carcinomas were also studied. This was followed by quantitative immunohistochemical staining for CD34, podoplanin, and Flt-4. Lymph node metastasis was significantly correlated with VEGF-D messenger RNA transcript levels (P=0.027) and VEGF-D immunoreactivity (P=0.019). Increased lymph vessel density was also correlated with VEGF-D expression and lymph node metastasis. In conclusion, our findings indicate that VEGF-D expression and increased lymph vessel density may have an important role for lymph node metastasis in papillary thyroid carcinoma. PMID- 15803189 TI - Absence of caspase 3 activation in neoplastic cells of nasopharyngeal carcinoma biopsies predicts rapid fatal outcome. AB - Poor prognosis in nasopharyngeal carcinoma patients may result from resistance to the apoptosis-inducing effect of radio- and/or chemotherapy. Apoptosis depends on proper activation of caspase 3, resulting in cleavage of key proteins like PARP 1. To investigate whether disruption of the apoptosis pathway results in therapy resistant tumour cells, we investigated whether absence of caspase 3 activation in tumour biopsies of nasopharyngeal carcinoma patients is related to poor clinical outcome. Moreover, we investigated whether absence of caspase 3 activation is related to loss of procaspase 3 expression or expression of the apoptosis regulators p53, bcl-2 and XIAP. We studied 36 Indonesian nasopharyngeal carcinoma patients without evidence of distant metastases who were treated with curative intent by radiotherapy only. Activation of caspase 3 and expression of the different markers were determined using specific antibodies. Levels of caspase 3 activation were determined by quantifying positively staining tumour cells. Nasopharyngeal carcinoma-derived C15 and C17 tumour cells were used as control. Absence of caspase 3 activation was strongly related to a poor clinical response to radiotherapy and to a higher T and N stage, resulting in a particularly poor clinical outcome with regard to progression-free (P<0.0001) and overall survival time (P<0.0001). Absence of caspase 3 activation was significantly correlated to loss of expression of procaspase 3 (P=0.04). In nasopharyngeal carcinoma patients treated with curative intent, absence of active caspase 3-positive neoplastic cells predicts rapid fatal outcome, and is associated with poor response to radiotherapy and high T and N stage at time of presentation. PMID- 15803190 TI - Intravascular large B-cell lymphoma involving hemangiomas: an unusual presentation of a rare neoplasm. AB - We report the clinicopathological features of two cases of intravascular large B cell lymphoma involving cutaneous hemangiomas. The cases were identified from the consultation files of two of the authors. Both patients were women, 64 and 55 years of age, who presented with long-standing cutaneous hemangiomas of the posterior scalp and left shoulder, respectively. The lesions were brought to medical attention by an increase in size and change in color. Biopsies and immunohistochemical evaluation of the hemangiomas revealed extensive involvement by intravascular large B-cell lymphoma. The neoplastic cells were diffusely positive for CD20 in both cases and negative for CD3, pan-cytokeratin (AE1/AE3), epithelial membrane antigen, S-100, Factor VIII-related antigen, CD34 and CD31. Disease was limited to the hemangiomas in both patients. Treatment consisted of chemotherapy (both patients) and adjuvant radiation therapy (one patient). One patient had a recurrence of disease 33 months after initial diagnosis, leading to an autologous stem cell transplant. The other patient is without evidence of disease 27 months after initial diagnosis. Although this is a rare neoplasm, it is important to consider intravascular large B-cell lymphoma in the differential diagnosis of vascular lesions containing intravascular neoplastic cells. PMID- 15803191 TI - Melanocytic lesions associated with dermatofibromas: a spectrum of lesions ranging from junctional nevus to malignant melanoma in situ. AB - Dermatofibromas are common lesions that are often associated with epidermal hyperplasia and basal layer hyperpigmentation. A single case of lentiginous melanocytic hyperplasia overlying a dermatofibroma has been reported, however, nevi and melanoma have to the best of our knowledge, not been previously reported. We present 14 cases of melanocytic lesions associated with dermatofibromas. The clinical data and hematoxylin- and eosin- stained sections were obtained and formalin-fixed, paraffin-embedded tissue was immunostained with antibodies against S-100, Mart-1, Factor XIIIa, and CD117. There were nine females and five males ranging in age from 30 to 64 years and anatomic sites included back (five), arm (six), flank (two), and leg (one). The clinical diagnosis ranged from dermatofibroma to desmoplastic melanoma. Histologically, the melanocytic lesions included junctional, compound, and dermal nevi, and malignant melanoma in situ. In four cases the dermal component appeared to merge with the dermatofibroma. In the case of the melanoma in situ, the dermatofibroma abutted the epidermis. Immunohistochemically, the melanocytic lesions were S-100/ Mart-1+, FXIIIa-, and the dermatofibromas were S-100/Mart-1-, FXIIIa+. Melanocytic neoplasia may appear in association with dermatofibromas. The fibrohistiocytic proliferation may be misinterpreted as a spindle or pleomorphic melanocytic process. Awareness of this association will aid in the correct diagnosis, and immunohistochemical studies will help in the differentiation of these two cell populations. PMID- 15803192 TI - Incipient chordoma: a report of two cases of early-stage chordoma arising from benign notochordal cell tumors. AB - Chordomas are rare malignant bone tumors primarily involving both ends of the axial skeleton that present as destructive bone lesions with a large soft tissue mass. Chordomas were previously believed to arise from notochordal remnants. However, recent studies suggest the possibility that chordomas arise from benign notochordal cell tumors. We present two cases of coccygeal incipient chordoma that strengthen the new hypothesis. The first case was an 83-year-old man who died of prostatic adenocarcinoma. The second case was a 79-year-old man who died of hepatocellular carcinoma. The coccygeal tumors were composed of intraosseous and extraosseous infiltrative lesions. The intraosseous lesions consisted of both benign notochordal cell tumor and incipient chordoma. The extraosseous lesions were consistent with incipient chordoma. In addition, two other small benign notochordal cell tumors were found at a different level in case 1. It is conceivable that pre-existing intraosseous benign notochordal cell tumors transform into incipient chordoma and then extend through the cortex into the surrounding soft tissue. The incidence of incipient chordoma appears much higher than expected because chordomas are rare tumors with an incidence of one case per 1 000 000 persons per year. We suspect that unknown factors transform incipient chordoma into classic chordoma. PMID- 15803193 TI - The evolving field of biodefence: therapeutic developments and diagnostics. AB - The threat of bioterrorism and the potential use of biological weapons against both military and civilian populations has become a major concern for governments around the world. For example, in 2001 anthrax-tainted letters resulted in several deaths, caused widespread public panic and exerted a heavy economic toll. If such a small-scale act of bioterrorism could have such a huge impact, then the effects of a large-scale attack would be catastrophic. This review covers recent progress in developing therapeutic countermeasures against, and diagnostics for, such agents. PMID- 15803194 TI - Protein drug stability: a formulation challenge. AB - The increasing use of recombinantly expressed therapeutic proteins in the pharmaceutical industry has highlighted issues such as their stability during long-term storage and means of efficacious delivery that avoid adverse immunogenic side effects. Controlled chemical modifications, such as substitutions, acylation and PEGylation, have fulfilled some but not all of their promises, while hydrogels and lipid-based formulations could well be developed into generic delivery systems. Strategies to curb the aggregation and misfolding of proteins during storage are likely to benefit from the recent surge of interest in protein fibrillation. This might in turn lead to generally accepted guidelines and tests to avoid unforeseen adverse effects in drug delivery. PMID- 15803195 TI - Chemistry-to-gene screens in Caenorhabditis elegans. AB - The nematode worm Caenorhabditis elegans is a genetic model organism linked to an impressive portfolio of fundamental discoveries in biology. This free-living nematode, which can be easily and inexpensively grown in the laboratory, is also a natural vehicle for screening for drugs that are active against nematode parasites. Here, we show that chemistry-to-gene screens using this animal model can define targets of antiparasitic drugs, identify novel candidate drug targets and contribute to the discovery of new drugs for treating human diseases. PMID- 15803196 TI - Osteoarthritis - an untreatable disease? AB - Osteoarthritis is a painful and disabling disease that affects millions of patients. Its aetiology is largely unknown, but is most likely multi-factorial. Osteoarthritis poses a dilemma: it often begins attacking different joint tissues long before middle age, but cannot be diagnosed until it becomes symptomatic decades later, at which point structural alterations are already quite advanced. In this review, osteoarthritis is considered as a disease of the whole joint that may result from multiple pathophysiological mechanisms, one of which is the dysregulation of lipid homeostasis. No proven disease-modifying therapy exists for osteoarthritis and current treatment options for chronic osteoarthritic pain are insufficient, but new pharmacotherapeutic options are emerging. PMID- 15803197 TI - Strategies for mapping and cloning quantitative trait genes in rodents. AB - Over the past 15 years, more than 2,000 quantitative trait loci (QTLs) have been identified in crosses between inbred strains of mice and rats, but less than 1% have been characterized at a molecular level. However, new resources, such as chromosome substitution strains and the proposed Collaborative Cross, together with new analytical tools, including probabilistic ancestral haplotype reconstruction in outbred mice, Yin-Yang crosses and in silico analysis of sequence variants in many inbred strains, could make QTL cloning tractable. We review the potential of these strategies to identify genes that underlie QTLs in rodents. PMID- 15803198 TI - Gene-environment interactions in human diseases. AB - Studies of gene-environment interactions aim to describe how genetic and environmental factors jointly influence the risk of developing a human disease. Gene-environment interactions can be described by using several models, which take into account the various ways in which genetic effects can be modified by environmental exposures, the number of levels of these exposures and the model on which the genetic effects are based. Choice of study design, sample size and genotyping technology influence the analysis and interpretation of observed gene environment interactions. Current systems for reporting epidemiological studies make it difficult to assess whether the observed interactions are reproducible, so suggestions are made for improvements in this area. PMID- 15803199 TI - Nuclear receptors--a perspective from Drosophila. AB - Nuclear receptors are ancient ligand-regulated transcription factors that control key metabolic and developmental pathways. The fruitfly Drosophila melanogaster has only 18 nuclear-receptor genes - far fewer than any other genetic model organism and representing all 6 subfamilies of vertebrate receptors. These unique attributes establish the fly as an ideal system for studying the regulation and function of nuclear receptors during development. Here, we review recent breakthroughs in our understanding of D. melanogaster nuclear receptors, and interpret these results in light of findings from their evolutionarily conserved vertebrate homologues. PMID- 15803200 TI - Fish as models for environmental genomics. AB - Fish offer important advantages for defining the organism-environment interface and responses to natural or anthropogenic stressors. Genomic approaches using fish promise increased investigative power, and have already provided insights into the mechanisms that underlie short-term and long-term environmental adaptations. The range of fish species for which genomic resources are available is increasing, but will require significant further expansion for the optimal application of fish environmental genomics. PMID- 15803201 TI - The Human Genome Diversity Project: past, present and future. AB - The Human Genome Project, in accomplishing its goal of sequencing one human genome, heralded a new era of research, a component of which is the systematic study of human genetic variation. Despite delays, the Human Genome Diversity Project has started to make progress in understanding the patterns of this variation and its causes, and also promises to provide important information for biomedical studies. PMID- 15803202 TI - Donor-substituted phenyl-pi-chromones: electrochemiluminescence and intriguing electronic properties. AB - Phenylethynylchromones bearing different donor groups at the phenyl moiety have been prepared and their photophysical and electrogenerated chemiluminescence (ECL) properties have been studied with respect to their structural features. Intriguingly, the presence and variation of donor groups do not much influence the absorption spectra, which can be compared with the spectrum of unsubstituted chromone, whereas the photoluminescence (PL) spectra show pronounced changes. Density functional theory (DFT) calculations indicate enhancement of HOMO energy levels upon increasing the donor strength. The photophysical properties have also been studied in various solvents, and the PL spectra in particular show the anticipated trend. The introduction of pi-extension imparts ECL to the new molecules and the electronic coupling between the donor and the acceptor moieties through C-C triple bond influences ECL emission maxima. Weaker donors impart excimer ECL while stronger donors impart monomeric intramolecular charge transfer (ICT) ECL. PMID- 15803203 TI - Optical properties of rhodoxanthin accumulated in Aloe arborescens Mill. leaves under high-light stress with special reference to its photoprotective function. AB - In Aloe arborescens Mill. leaves, strong sunlight or its combination with drought induces the accumulation of the red keto-carotenoid, rhodoxanthin. Simultaneously, the transformation of chloroplasts into chromoplasts accompanied by degradation of thylakoid membranes and formation of plastoglobuli, large in size and number, takes place. Depending on stress conditions the build up of rhodoxantin occurred along with the loss of chlorophyll or on the background of relatively high content of the pigment in the leaves. Microspectrophotometrical measurements showed the presence of chlorophyll-free plastids and retention of carotenoids during leaf adaptation to strong sunlight. The plastid spectra contained absorption bands of common for higher plants carotenoids together with those of rhodoxantin, with absorption maxima situated in the blue (440-480 nm) and the green ranges of the spectrum, respectively. The studies of whole-leaf optical properties revealed a broad band of rhodoxanthin absorption in the blue green range peaking near 540-550 nm. Within this spectral band the accumulation of rhodoxanthin occurring, probably, in plastoglobuli considerably increased light absorption by stressed Aloe leaves. A possible photoprotective function of rhodoxanthin and other carotenoids as an internal light trap analogous to that accomplished by anthocyanins in other plant species is discussed. PMID- 15803204 TI - Transgene delivery and gelonin cytotoxicity enhanced by photochemical internalization in fibroblast-like synoviocytes (FLS) from rheumatoid arthritis patients. AB - The objective of this study was to determine if photochemical internalization (PCI) of gelonin can improve the treatment outcome as compared to photodynamic therapy (PDT) and gene transduction of fibroblast-like synoviocytes (FLS)in vitro. For this purpose synovial tissue was obtained under synovectomy of rheumatoid arthritis (RA) patients. Primary single cell suspensions were treated with the photosensitizer meso-tetraphenylporphine (TPPS2a) and light exposure (PDT) followed by evaluation of the cell survival by flow cytometry. PCI of gelonin was performed on FLS in passages 4 and 5 after removal from patients followed by measurements of protein synthesis 24 h after treatment. Additionally FLS were transduced with an adenovirus encoding the E.coli. lacZ gene and treated with PCI to evaluate the effect on the transduction rate. As a result all the cells in the primary cell suspension were susceptible to PDT but CD 106- (FLS) and CD14-positive (monocytes) cells were more sensitive to inactivation by PDT than CD2- (T-cells) and CD19-positive (B-cells) cells. With respect to protein synthesis FLS became up to 4-fold more sensitive to light when combining the photochemical treatment with the gelonin incubation. The fraction of virally transduced FLS was approximately doubled by means of PCI. In conclusion our experiments showed that PCI increased the cytotoxic effect of gelonin and adenoviral transduction of FLS derived from RA patients. PMID- 15803205 TI - Combinations of chiral and prochiral singlet radical-pairs in reaction cavities of polyethylene films. Control and analysis of radical tumbling and translation. AB - The regio- and stereo-chemistries of combination products from chiral 1 naphthoxy/(R)-2-phenylpropanoyl and prochiral 1-naphthoxy/1-phenylethyl singlet radical-pairs (radical-pairs A and B, respectively) have been studied at different temperatures in polyethylene (PE) films with different crystallinities. The radical-pairs have been generated as intermediates along the photo-Fries reaction course of 1-naphthyl (R)-2-phenylpropanoate ((R)-1) and the photo Claisen reaction course of 1-naphthyl (R)-1-phenylethyl ether ((R)-2). Radical pair was produced directly upon lysis of the first excited singlet state of (R)-2 and indirectly after irradiation of (R)-1 and subsequent decarbonylation of the 2 phenylpropanoyl radical of radical-pair A. Comparison of the fates of the directly and indirectly formed radical-pairs provides detailed information about the nature of the reaction cavities within the polyethylene hosts and how the combinations of the radical-pairs are influenced by their initial locations within a cavity. The results, especially when taken with those from irradiations in n-alkanes, indicate that the cavities are "templated" by the (R)-1 and (R)-2 guest molecules and that the templated shapes are retained in some form for periods that are at least as long as the time required for decarbonylation of a 2 phenylpropanoyl radical. In addition, the enantiomeric excesses of the decarbonylated photoproducts from (R)-1(2, 2-(1-phenylethyl)-1-naphthol (2BN), and 4-(1-phenylethyl)-1-naphthol (4BN)) or 2BN and 4 BN from (R)-2 indicate different influences of temperature on translational and tumbling motions of the radicals of radical-pairs B within their polyethylene cages. PMID- 15803206 TI - Conduction and photoelectrochemical properties of monomeric and electropolymerized tetraruthenated porphyrin films. AB - The influence of the preparation method on the structure, conduction and photoelectrochemical properties of monomeric and polymeric tetraruthenated porphyrin films on ITO glass and nanocrystalline TiO2 has been investigated. The films were characterized by STM, MAC mode SFM, cyclic voltammetry, electrochemical impedance spectroscopy (EIS) and combined electro /photoelectrochemical techniques. The electronic diffusion coefficient D(e)C(m)2 of the films differed by three to four orders of magnitude depending on the procedure employed for the deposition process. The photoelectrochemical properties were evaluated either: by depositing the films directly on transparent ITO electrodes, under an applied bias potential and presence of O2 as electron acceptor; or by depositing the porphyrin material on nanocrystalline TiO2 in a Gratzel-type cell. In the first case the porphyrin films exhibited a typical p type semiconductor behavior described by a Schottky junction model, while in the second the films behaved as a sensitizer of an n-type semiconductor. The photoelectrochemical properties of the porphyrin films and their performance as sensitizer in Gratzel-type cells were found to be strongly dependent on the conductivity and packing characteristics of the material. Semi-empirical calculations were performed by modified MM2 and ZINDO/S methods, in order to simulate the packing and electronic structures of the tetraruthenated porphyrin. PMID- 15803207 TI - Intramolecular and intermolecular hydrogen-bonding effects on photophysical properties of 2'-aminoacetophenone and its derivatives in solution. AB - Effects of intra- and intermolecular hydrogen-bonds on the photophysical properties of 2'-aminoacetophenone derivatives (X-C6H4-COCH3) having a substituted amino group (X) with different hydrogen-bonding ability to the carbonyl oxygen (X: NH2(AAP), NHCH3(MAAP), N(CH3)2(DMAAP), NHCOCH3(AAAP), NHCOCF3(TFAAP)) are investigated by means of steady-state and time-resolved fluorescence spectroscopy and time-resolved thermal lensing. Based on the photophysical parameters obtained in aprotic solvents with different polarity and protic solvents with different hydrogen-bonding ability, the characteristic photophysical behavior of the 2'-aminoacetophenone derivatives is discussed in terms of hydrogen-bonding and n,pi*-pi,pi* vibronic coupling. The dominant deactivation process of AAP and MAAP in nonpolar aprotic solvents is the extremely fast internal conversion (k(ic)= 1.0 x 10(11) s(-1) for AAP and 3.9 x 10(10) s(-1) for MAAP in n-hexane). The internal conversion rates of both compounds decrease markedly with increasing solvent polarity, suggesting that vibronic interactions between close-lying S1(pi,pi*) and S2(n,pi*) states lead to the large increase in the non-radiative decay rate of the lowest excited singlet state. It is also suggested that for MAAP, which has a stronger hydrogen-bond as compared to AAP, an intramolecular hydrogen-bonding induced deactivation is involved in the dissipation of the S1 state. For DMAAP, which cannot possess an intramolecular hydrogen-bond, the primary relaxation mechanism of the S1 state in nonpolar aprotic solvents is the intersystem crossing to the triplet state, whereas in protic solvents very efficient internal conversion due to intermolecular hydrogen-bonding is induced. In contrast, the fluorescence spectra of AAAP and TFAAP, which have an amino group with a much stronger hydrogen bonding ability, give strongly Stokes-shifted fluorescence, indicating that these compounds undergo excited-state intramolecular proton transfer reaction upon electronic excitation. PMID- 15803208 TI - Short- and long-term effects of solar ultraviolet radiation on the red algae Porphyridium cruentum (S. F. Gray) Nageli. AB - During spring 2002 and fall 2003 we carried out experiment in tropical southern China to determine the short- and long-term effects of solar ultraviolet radiation (UVR, 280-400 nm) on photosynthesis and growth in the unicellular red alga Porphyridium cruentum. During the experimentation, cells of P. cruentum were exposed to three radiation treatments: (a) samples exposed to PAR (400-700 nm) + UV-A (315-400 nm) + UV-B (280-315 nm)(PAB treatment); (b) samples exposed to PAR + UV-A (PA treatment) and, (c) samples exposed only to PAR (P treatment). To assess the short-term impact of UVR as a function of irradiance, we determined photosynthesis versus irradiance (Pvs.E) curves. From these curves the maximum carbon uptake rate (P(max)) and the light saturation parameter (E(k)) were obtained, with values of approximately 12.8-14.4 microg C (microg chl a)(-1) h( 1), and approximately 250 micromol m(-2) s(-1), respectively. A significant UVR effect on assimilation numbers was observed when samples were exposed at irradiances higher than E(k), with samples exposed to full solar radiation having significant less carbon fixation than those exposed only to PAR. Biological weighting functions of P. cruentum were used to evaluate the UVR impact per unit energy received by the cells; the data indicate that the species is as sensitive as natural phytoplankton from the southern China Sea; however, it is much more resistant than Antarctic assemblages. When evaluating the combined effects of mixing speed and UVR, it was seen that samples rotating fast within the upper mixed layer were less inhibited by UVR as compared to those under slow mixing or in fixed samples. Growth of P. cruentum over a week-long experiment was not affected by neither UVR nor UV-A; additionally, low photoinhibition was found at the end as compared to that at the beginning of this experiment. Our results thus indicate that, although on short-term basis P. cruentum is affected by solar UVR, it can acclimate to minimize UVR-induced effects when given enough time. PMID- 15803209 TI - Cell specific effects of polyunsaturated fatty acids on 5-aminolevulinic acid based photosensitization. AB - The purpose of this study was to examine whether the dietary components n-6 and n 3 polyunsaturated fatty acids (PUFAs) may potentiate the effect of photodynamic therapy (PDT) in human cancer cell lines by enhancing the lipid peroxidation. The effects of the porphyrin precursor 5-aminolevulinic acid (5-ALA) and light (320 < lambda < 440 nm, 33 W m(-2)), with or without docosahexaenoic acid (DHA) or arachidonic acid (AA), were tested in the colon carcinoma cell lines SW480 and WiDr, the glioblastoma cell line A-172 and the lung adenocarcinoma cell line A 427. The production of endogenous protoporphyrin IX (PpIX) varied substantially between the cell lines and was approximately 4-fold higher in WiDr as compared with SW480. Cell killing by 5-ALA-PDT also varied between the cell lines, but without clear correlation with PpIX levels. Treatment with DHA or AA (10 or 70 microM, 48 or 72 h) in combination with 5-ALA-PDT (1 or 2 mM) enhanced the cytotoxic effect in A-172 and A-427 cells, but not in SW480 and WiDr cells. While 5-ALA-PDT alone increased the lipid peroxidation in A-172 and WiDr cells only, 5 ALA-PDT plus PUFAs increased the lipid peroxidation substantially in all four cell lines. Interestingly, alpha-tocopherol (50 microM, 48 h) strongly reduced lipid peroxidation after all treatments in all cell lines, while cytotoxicity was only reduced substantially in A-427 cells. This demonstrates that induction of lipid peroxidation is not a general mechanism responsible for the cytotoxicity of 5-ALA-PDT, although it may be important in cell lines with an inherent sensitivity to lipid peroxidation products. Thus, the mechanisms of cell growth inhibition/cell killing by PDT are complex and cell specific. PMID- 15803210 TI - Abstracts of the 5th European Congress on Clinical and Economic Aspects of Osteoporosis and Osteoarthritis, 16-19 March 2005, Rome, Italy. PMID- 15803211 TI - [Regenerative medicine and plastic surgery]. AB - Regenerative medicine recently evolved as a new medical field that includes tissue engineering, cell/system biology, nanotechnology, pharmacology, stem-cell biology, and bioengineering. Regenerative medicine targets new forms of therapy to promote and support the intrinsic, autologous, regenerative potential of human biological systems. All fields of surgery have profited from these developments, and spectacular experimental results and clinical benefits have been obtained. Plastic surgery has shown interest in regenerative medicine due to its focus on reconstructive surgery. Early on, several interdisciplinary experimental working groups were founded including plastic surgery. This overview takes a closer look at common experimental and clinical results of regenerative medicine and plastic surgery. PMID- 15803212 TI - Hox11L1 expression by precursors of enteric smooth muscle: an alternative explanation for megacecum in HOX11L1-/- mice. AB - Previous studies have focused on expression of Hox11L1 in enteric neurons as the explanation for intestinal and urinary bladder dysmotility observed in mice that do not have the transcription factor. However, Hox11L1 is also expressed transiently in endo-, meso-, and ectodermal cells of the most caudal embryo during gastrulation. We sought to more fully characterize the fates of these cells because they might help explain the pathogenesis of lethal pseudo obstruction in Hox11L1-null mice. The Cre recombinase cDNA was introduced into the Hox11L1 locus, and expression of the "knock-in" allele was used to activate the Rosa26R, beta-galactosidase reporter gene in cells with ongoing Hox11L1 transcription and their descendants. During gastrulation, Rosa26R activation was observed in progenitors of caudal somatic and visceral cells, including enteric smooth muscle. Expression in enteric neural precursors appeared much later. Analysis of endogenous Hox11L1 mRNA in aneuronal segments of large intestine that were grafted under the renal capsule indicated that the early activation of Hox11L1 in visceral mesoderm was transient and ceased before colonization of the large intestine by neural progenitors. Mice homozygous for the Cre allele died shortly after weaning, with cecal and proximal colonic distention but without overt anatomic defects that might represent maldevelopment of the visceral mesoderm. Our findings expand the range of possible functions of Hox11L1 to include activation of an as yet unknown developmental program in visceral smooth muscle and allow the possibility that intestinal dysmotility in Hox11L1-null animals may not be a primary neural disorder. PMID- 15803213 TI - Sudden, unexpected death associated with meningioangiomatosis: case report. AB - We report a case of sudden, unexpected death associated with meningioangiomatosis in a 13-year-old, previously healthy male without a history of seizures, neurologic deficits, or clinical stigmata of neurofibromatosis. There was no family history of neurofibromatosis. The postmortem examination showed a 5-cm mass involving the right posterior frontal and orbital frontal cortex that had microscopic features diagnostic of meningioangiomatosis. Because no other cause of death was found, we postulate that he likely died as a result of a seizure secondary to meningioangiomatosis. PMID- 15803214 TI - Tumors associated with sudden infant and childhood death. AB - Tumors are rare causes of sudden death in infancy and early childhood. The goals of this study were to determine the types and frequency of the tumors associated with sudden death occurring in cases between birth and age 3 years. The San Diego Sudden Infant Death Syndrome/sudden unexplained death in childhood (SUDC) Research Project database and the literature were reviewed retrospectively. Sixty eight cases, with the most (84%) affecting the heart and brain, were identified. Tumors are a rare but significant cause of sudden death in infancy and early childhood, and their diagnosis may have significant genetic implications for planning future pregnancies. The diagnosis of these lesions can be established only after thorough postmortem examination. PMID- 15803215 TI - Testicular mixed germ cell tumor with polyembryoma component in brothers. AB - We report the case of a 17-year-old male with a testicular tumor and high serum levels of alpha-fetoprotein. The patient was treated with surgery followed by combination chemotherapy with bleomycin, etoposide, and cisplatin. Histologic examination showed features of a mixed germ cell tumor composed of mature teratoma, immature teratoma, embryonal carcinoma, yolk sac tumor, and polyembryoma. He is currently well, and his serum levels of alpha-fetoprotein have been normal more than 5 months after treatment. His brother, aged 17 years at the time, had a similar tumor removed from the right testicle 5 years previously. PMID- 15803216 TI - Perivascular epithelioid cell tumor of the orbit: a case report and review of the literature. AB - We present a unique case of a perivascular epithelioid cell tumor (PEComa) in the orbit of a 9-year-old female patient. The entity of PEComas has been described only recently. Characteristic histologic features and an immunohistochemical profile of negativity for epithelial markers and positivity for melanogenesis related markers define the tumors. In children and young adults, this tumor has a predilection for the falciform ligament and ligamentum teres of the liver. It is associated with, but not exclusive to, tuberous sclerosis. To the best of our knowledge, this is the first reported case of a PEComa of the orbit in a child or adult. The main differential diagnoses for this melanin pigment-producing lesion include melanoma and pigmented paraganglioma. The histologic features, immunohistochemical profile, ultrastructural studies, and molecular studies led us to favor a diagnosis of PEComa. The prognosis of this entity is undetermined due largely to the small number of reported cases. PMID- 15803217 TI - "Proximal-type" and classic epithelioid sarcomas represent a clinicopathologic continuum: case report. AB - We report a case of an epithelioid sarcoma that occurred in the right hand of a 14-year-old boy and had the "proximal-type" morphology and a complex, near tetraploid karyotype. The tumor metastasized to the lungs, where the morphology was typical for the classic epithelioid sarcoma. Based on the morphologic and cytogenetic findings in this case, we suggest that the proximal-type and the classic epithelioid sarcomas are not distinct entities but represent a continuum. PMID- 15803218 TI - Mosaic duplication 1(q11q44) in an infant with nephroblastomatosis and mineralization of extraplacental membranes. AB - Partial trisomy of 1q is rare. Only 32 cases of isolated partial trisomy 1q have been previously reported. From these cases, a characteristic phenotype is beginning to emerge. We present a case of mosaic duplication of 1q [46,XX,dup (1)(q11q44)/46,XX]. Many features of our patient have been described in previous patients, thus supporting the emerging phenotype. Two particular features, however, have not been previously described. The present case demonstrated extensive mineralization of the extraplacental membranes and bilateral nephromegaly, with an extreme form of diffusely hyperplastic perilobar nephroblastomatosis. Clinical comparison is made between our case and previously reported cases, and the clinical significance of the unique findings are reviewed and discussed. PMID- 15803220 TI - Sclerosing rhabdomyosarcomas in children and adolescents: a clinicopathologic review of 13 cases from the Intergroup Rhabdomyosarcoma Study Group and Children's Oncology Group. PMID- 15803222 TI - Bowel obstruction due to Littre hernia: CT diagnosis. AB - Meckel diverticulum is the commonest congenital anomaly of the gastrointestinal tract. Any hernia containing the Meckel diverticulum is termed a Littre hernia. Littre hernias are commonest in the inguinal region and may cause bowel obstruction secondary to strangulation or incarceration of the diverticulum within the hernial sac. The diagnosis of an incarcerated Littre hernia on computed tomogram has not been previously reported in the scientific literature. PMID- 15803223 TI - Enterocele associated with rectocele revealed by dynamic pelvic CT. AB - Enterocele is often associated with other pelvic floor disorders but it is not always possible to detect by clinical examination. Defecography with peritoneography and/or barium meal intake has recently been developed as a new method to identify enterocele, but this method is an invasive procedure. Multislice computed tomography was performed at rest and during simulated defecation to evaluate an 80-year-old female patient who had a defecation disorder and was diagnosed as having rectocele based on results from defecography and clinical findings. Multiplanar reconstruction images were generated for image evaluation. Using this novel method of dynamic pelvic computed tomography, a third-degree enterocele was clearly demonstrated in this case. PMID- 15803224 TI - Spigelian hernia with unusual content. AB - We present a case of a primary tumor of the peritoneum that manifested as a spigelian hernia in a 74-year-old woman. Multidetector computed tomography showed a large heterogeneous mass located subcutaneously on the right spigelian line connected to the abdominal cavity. We found no previous report describing a primary peritoneal tumor in a spigelian hernia. PMID- 15803225 TI - Acute appendicitis within a femoral hernia: multidetector CT findings. AB - We present a case of surgically proved acute appendicitis strangulated in a femoral hernia sac. Multidetector computed tomography with multiplanar reformations was thought to be helpful in evaluating acute appendicitis in a femoral hernia sac. An awareness of computed tomographic findings may avoid a delay in recognition and management of a patient with this unusual condition. PMID- 15803226 TI - Radiofrequency ablation: other abdominal organs. PMID- 15803227 TI - Disruptive visions: predictive simulation--between scientific method and clinical trial is the role of modeling and simulation in scientific discovery and validation. AB - The foundations of surgery have rested upon the passing of knowledge from generation to generation by mentors and manuscripts. Until the 1900 s, the passage of knowledge was through tradition. Since then, it has changed from observation and experience to the scientific method to clinical trials. A new approach, derived from other scientific disciplines, is that of predicting results from modeling and simulation, which will allow for acceleration of the process of validation of discoveries and optimizing the implementation of clinical trials in order to more rapidly transfer trusted knowledge from generation to generation. PMID- 15803228 TI - Endoscopic treatment modalities for gastroesophageal reflux disease. AB - A debate has been going for decades between surgeons and gastroenterologists about the treatment of choice for gastroesophageal reflux disease (GERD). The lower esophageal sphincter (LES) has been historically far from the reach of gastroenterologists, who adopted the symptomatic treatment as their approach to reflux disease through reduction of gastric acid. As for surgeons, reaching the LES was only possible by invading the thoracic or abdominal cavity. Although their approach was later refined to become "minimally" invasive, it was still deemed too invasive by others to allow it to be the "gold standard." Simple logic should lead one to think about the "natural route" as the easiest way to reach the LES. This concept has opened the door for the new era of GERD treatment through "endoscopic modality." Seven different techniques are currently being used to treat patients with GERD. We review the mechanism of action, potential side effects, efficacy, durability, and results from the most recent or largest experience of each. This review shows that endoscopic treatment has definitely earned its place as a viable option for GERD treatment in selected patients. With the available data from clinical trials, it is not possible to determine the best modality available, and the endoscopic treatment of choice is to be determined with further studies. PMID- 15803229 TI - Early versus delayed laparoscopic cholecystectomy for acute cholecystitis: a prospective randomized trial. AB - BACKGROUND: The role of laparoscopic cholecystectomy for acute cholecystitis is not yet clearly established. The aim of this prospective randomized study was to evaluate the safety and feasibility of laparoscopic cholecystectomy for acute cholecystitis and to compare the results with delayed cholecystectomy. METHODS: Between January 2001 and November 2002, 40 patients with a diagnosis of acute cholecystitis were assigned randomly to early laparoscopic cholecystectomy within 24 h of admission (early group, n = 20) or to initial conservative treatment followed by delayed laparoscopic cholecystectomy, 6 to 12 weeks later (delayed group, n = 20). RESULTS: There was no significant difference in the conversion rates (early, 25% vs delayed, 25%), operating times (early, 104 min vs delayed, 93 min), postoperative analgesia requirements (early, 5.3 days vs delayed, 4.8 days), or postoperative complications (early, 15% vs delayed, 20%). However, the early group had significantly more blood loss (228 vs 114 ml) and shorter hospital stay (4.1 vs 10.1 days). CONCLUSIONS: Early laparoscopic cholecystectomy for acute cholecystitis is safe and feasible, offering the additional benefit of a shorter hospital stay. It should be offered to patients with acute cholecystitis, provided the surgery is performed within 72 to 96 h of the onset of symptoms. PMID- 15803230 TI - The impact of a resident's seniority on operative time and length of hospital stay for laparoscopic appendectomy: outcomes used to measure the resident's laparoscopic skills. AB - BACKGROUND: Laparoscopic appendectomy (LA) frequently is performed by residents during calls. This study aimed at evaluating residents' surgical skills using parameters of operating time, length of hospital stay (LOS), and conversion rate in correlation with the operating team's level of seniority. In addition, this study compared the operating time for LA with that for open appendectomy performed by the same teams, and identified deterministic factors that have an impact on such parameters. METHODS: All records of patients undergoing appendectomy performed by residents alone during a 32-month period were reviewed retrospectively. Eight residents were assigned to two levels of seniority: juniors 3 years (S). Operating time and LOS were compared between the three surgical teams, namely, J/J, J/S, and S/J as operating and assistant surgeons, respectively. Operating time, conversion rates, and LOS were compared for the same team combinations. RESULTS: Residents alone performed 341 (151 laparoscopic and 190 open) appendectomies during on-call hours. Four of the residents had been 3 years or less in residency (J), and four had been in residency more than 3 years (S). The overall mean operating time was 1.33 +/- 0.48 h for LA and 1.2 +/- 0.5 h for open appendectomy (p = 0.016). The operating time correlated with the level of training for both LA (J/J, 1.6 +/- 0.38 h; J/S, 1.41 +/- 0.37 h; S/J, 1. 25 +/- 0.4 h; p = 0.03, ANOVA) and open appendectomy (J/J, 1.53 +/- 0.89 h; J/S, 1.4 +/- 0.63 h; S/J, 0.86 +/- 0.45 h; p = 0.023, ANOVA). The mean LOS was 2.9 +/- 3.1 days for open appendectomy and 2.1 +/- 2.8 days for LA (p = 0.065), and was not different after operation by any of the teams (J/J, J/S, S/J) for either the open or the laparoscopic procedure. CONCLUSIONS: There is a distinct difference in the surgical skills of residents according to level of seniority, as primarily reflected by operating time. Laparoscopic appendectomy requires longer time to perform in a teaching setting, but the most deterministic factor that dictates operating time is the composition of the surgical team rather than the laparoscopic approach. PMID- 15803231 TI - Outpatient laparoscopic splenectomy: patient safety and satisfaction. AB - BACKGROUND: We assessed the feasibility of outpatient laparoscopic splenectomy, as performed by an experienced laparoscopic term and combined with optimal anesthesia. METHODS: Inclusion criteria in the study was limited to patients not hospitalized before the procedure who had hematological or neoplastic indications for splenectomy and were classified as American Society of Anesthesiologists (ASA) I-III. They received general intravenous anesthesia with propofol and remifentanil and were given keterolac, propacetamol, droperidol, and ondansetron as prophylaxis against postoperative pain and nausea. Laparoscopic splenectomy was performed via three trocars. The specimen was removed via an incision in the left iliac fossa. RESULTS: Ten of the 12 patients were discharged 3-6 h postoperatively; the other two were admitted primarily to hospital. One was readmitted due to a fever, which was finally explained by measles. The median operative times was 58 min (range, 45-135). Patient satisfaction was excellent in nine and intermediate in two cases; it was poor in one case, due to postoperative pain. CONCLUSION: Laparoscopic splenectomy can be completed in a relatively short time; therefore, it is feasible, safe, and satisfactory for most patients as an outpatient procedure. PMID- 15803232 TI - Laparoscopic hand-assisted spleen autotransplantation. AB - BACKGROUND: Only recently has the spleen been perceived as an organ with a major immune function. This raised an interest in spleen salvage after spleen trauma and pancreatic tail resection, for the treatment of hematologic disorders and inducement tolerance for allogenic transplants. The purpose of this study was to evaluate the feasibility of a new technique for spleen transplantation: laparoscopic spleen autotransplantation in a large animal model. METHODS: Ten 35 kg pigs were used for this study. A laparoscopic hand-assisted splenectomy was first performed. The spleen was extracted through the handport to be flushed with a 4 degrees C saline solution and prepared extracorporeally. The graft was then reintroduced into the same animal's abdominal cavity, and a splenic-to-common iliac artery and vein bypass was performed laparoscopically using a 7-0 polytetrafluoroethylene running suture. The animal was killed 1 week postoperatively for histologic examination. RESULTS: All 10 animals tolerated the procedure well. No conversion to open surgery was required. The mean operative time was 253 +/- 45 min. The mean time needed to create the artery and vein anastomoses was 116 +/- 165 min, and the mean blood loss was 190 +/- 120 ml. There was no intra- or postoperative death. Intraoperative complications included two stenosed vascular anastomoses, which were taken down and revised. Seven of the 10 spleens were histologically viable 1 week after surgery. The nonviable transplantations were attributable to a thrombosis of the common iliac artery (n = 1) or the transplant artery (n = 2). CONCLUSIONS: Hand-assisted laparoscopic spleen autotransplantation is feasible in an animal model. This procedure could constitute an option when spleen resection is necessary for pancreatic tail resection, or when spleen preservation is important to the maintenance or restoration of an immune function. PMID- 15803233 TI - Hand-assisted laparoscopic splenectomy in the setting of splenomegaly. AB - BACKGROUND: Hand-assisted laparoscopic surgery (HALS) devices may be well suited to splenectomy in cases of splenomegaly. METHODS: All cases of HALS for splenectomy between 1997 and 2001 were reviewed. Patient characteristics, operative details, and morbidity and mortality were analyzed. RESULTS: HALS for splenectomy was performed in 54 patients. A total of 39 patients with massive splenomegaly (MS) (>600 g) were identified. The average weight of the MS group was 1285 +/- 505 g. There was one (3%) conversion. Operative time was 159 +/- 65 min, estimated blood loss was 257 +/- 240 ml, and length of hospital stay was 5.4 +/- 2.9 days. Morbidity was limited to 13 patients (24%), and there were two postoperative mortalities (5.1%). CONCLUSIONS: HALS for splenectomy in the setting of splenomegaly is feasible and safe. For the surgeon considering a laparoscopic approach in the setting of splenomegaly, a hand-assisted technique is ideally suited for removal of the enlarged spleen. PMID- 15803234 TI - Laparoscopic treatment of sigmoid diverticulitis: a retrospective review of 103 cases. AB - BACKGROUND: Laparoscopic treatment of sigmoid diverticulitis is commonly accepted in Hinchey cases I and II, whereas it is debated in the case of purulent peritonitis, and not indicated for fecal peritonitis. METHODS: A single-center experience of 103 patients treated for Hinchey I-III sigmoid diverticulitis was reviewed. One-stage laparoscopic resection and primary anastomosis constituted the planned procedure. Abscesses in patients with Hinchey IIa were drained percutaneously before surgery. Patients with Hinchey III underwent surgery in emergency. A four-trocar approach with left iliac fossa minilaparotomy was used. Fistulas were treated laparoscopically with Harmonic Scalpel dissection. RESULTS: Laparoscopic treatment was successfully completed for 100 patients. Intraoperative complications occurred in 2.9% of the cases. Postoperative procedure-related morbidity was 8%, occurring mainly in Hinchey I patients. A longer hospital stay was recorded among Hinchey IIb patients treated for colovescical fistula. No mortality was observed. CONCLUSIONS: Laparoscopic surgery for sigmoid diverticulitis in experienced hands can be a safe and effective gold standard procedure also for patients with fistula or purulent peritonitis. PMID- 15803235 TI - Midterm follow-up study of laparoscopic dissection of uterine vessels for surgical treatment of symptomatic fibroids. AB - BACKGROUND: This study aimed to assess laparoscopic dissection of uterine vessels (LDUV) for symptomatic fibroids in women. METHODS: A total of 69 women entered the study between March 2000 and June 2003. In this case series, 68 consecutive women underwent LDUV using ultrasonically activated sheers or electrosurgery for the treatment of fibroids over 3 years (median follow-up period, 14.5 months). Ultrasound or magnetic resonance imaging was carried out 3, 6, 12, 24, and 36 months after treatment. The tissue markers, gonadotropin, and estrogen levels were studied postoperatively. RESULTS: Almost all the patients (98.5%) had a successful LDUV with a low rate (7.3%) of postoperative complications. The time of surgery ranged from 15 to 50 min (mean, 30.8 min). The blood loss was minimal (mean, 14.7 ml), and the hospital stay was 2.4 days. Symptom improvement (menorrhagia or dysmenorrhoea) was 93.2%, and the average reduction in the dominant myoma was 57.8% during a follow-up period longer than 12 months. All the patients with anemia had normal red cell counts after 3 months. CONCLUSIONS: Uterine volume and the dominant fibroid were significantly reduced and symptoms were improved by LDUV. The laparoscopic procedure is associated with insignificant tissue damage and normal gonadotropin and estrogen levels. PMID- 15803236 TI - Microcirculation and excretory function of the liver under conditions of carbon dioxide pneumoperitoneum. AB - BACKGROUND: To date, the effects of increased abdominal pressure, as given during carbon dioxide (CO(2)) pneumoperitoneum, on hepatic microcirculation and biliary excretion are unknown. METHODS: Using a custom-made peritoneal cavity chamber, we performed intravital microscopy of the left liver lobe under conditions of CO(2) pneumoperitoneum in a rat model. In addition, biliary excretion was assessed. RESULTS: The establishment of a CO(2) pneumoperitoneum of 4 or 8 mmHg resulted in sinusoidal perfusion failure that was more pronounced in the periportal regions than in the midzonal and pericentral regions of the liver acinus. Biliary excretion was considerably reduced at an intraabdominal pressure of 8 mmHg. Leukocyte-endothelial cell interactions increased significantly in both hepatic sinusoids and postsinusoidal venules. CONCLUSION: Alterations in hepatic microcirculation and liver function must be taken into consideration in any kind of laparoscopic surgery and may be of particular clinical relevance in patients with liver pathology. PMID- 15803237 TI - Pulmonary sequestration children: is the thoracoscopic approach a good option? AB - BACKGROUND: The thoracoscopic approach for pulmonary sequestration in children seems to be feasible and reproducible. To date, there have been no studies comparing the thoracoscopic approach with the conventional technique in this indication. Therefore, the aim of this study was to compare these two approachs. METHODS: We performed a retrospective study in patients treated surgically for a prenatal diagnosis of pulmonary sequestration between 2000 and 2003. Three or five ports were used, and single-lung ventilation was required. The clinical, operative, and postoperative data were compared with those for patients operated on during the same period via thoracotomy. For statistical purposes, the children who required conversion to the open proecedure were grouped with those who underwent thoracotomy. The following data were compared: age, and weight, operating time, duration of postoperative drainage via chest tube, duration of postoperative narcotic use, pain scores on the 1(st) and 2(nd) days after the procedure, and length of hospital stay. The Mann-Whitney test was used for statistical analysis. RESULTS: Five infants were operated on via a thoracoscopic approach. Two required conversion to an open procedure. There were four intralobar sequestrations (ILS) and one extralobar sequestration (ELS). The mean age at surgery was 6.2 months (range, 4-9). The mean operating time, excluding the patients who were converted to open, was 155 min (range, 120-190). In the thoracotomy group, there were one ELS and one ILS. Two groups were defined: the exclusively laparoscopy group (EL) (n = 3) and the thoracotomy and conversion group (TC) (n = 4). The duration of postoperative drainage, duration of postoperative narcotic use, and hospital stay seemed to be shorter in the group EL. These times, respectively, were 3.4 vs 2.75 days, 4.1 vs 3.3 days, and 7.5 vs 4.3 days = (p = 0.03). There were two complications in the TC group (one wound abcess and one pleural leak that required a chest tube). CONCLUSION: The thoracoscopic approach for pulmonary sequestration is feasible. It seems to reduce the duration of postoperative drainage, postoperative narcotic use, hospital and stay. These results need to be confirmed by a prospective and more detailed study. PMID- 15803238 TI - Low-pressure pneumoperitoneum combined with intraperitoneal saline washout for reduction of pain after laparoscopic cholecystectomy: a prospective randomized study. AB - BACKGROUND: We designed a prospective randomized clinical trial to investigate whether intraperitoneal saline washout combined with a low-pressure pneumoperitoneum (LPSW) was superior to low-pressure pneumoperitoneum (LP) alone as a means of reducing postoperative pain and analgesic consumption in the early recovery period after laparoscopic cholecystectomy (LC). METHODS: A total of 124 consecutive patients undergoing LC due to uncomplicated symptomatic gallstones were randomized to the LP or LPSW group. In the LPSW group, normal saline at body temperature (25 ml/kg of body weight) was irrigated under the diaphragm. The fluid was evacuated via the passive-flow method through a 16-F closed drain left under the liver for 24 h. We then assessed the intensity of total abdominal postoperative pain using the Visual Analogue Scale (VAS), including the incidence of shoulder-tip pain (STP), total daily analgesia demand rate, analgesic consumption. Quality of life (QOL) within 7 days after the operation was assessed using the Medical Outcomes Study Short Form 36 Health Survey (SF-36). A p value of <0.05 was considered significant. RESULTS: The mean postoperative pain score was lower by 2.64 +/- 0.86 in the LPSW; the difference equaled 9.64% (p < 0.05). The incidence of STP was lower in the LPSW group (LP 11.29% vs LPSW 1.6%; p = 0.028). The analgesia demand rate was remarkably lower in LPSW vs LP within 24 and 48 h postoperatively (70.96% vs 90.32%; p = 0.006 and 64.51% vs. 83.87%; p = 0.013, respectively). After LPSW vs LP, QOL was better in terms of physical functioning, role limitations due to physical problems, and bodily pain (90.32% vs 77.42%; p = 0.05, 90.32% vs 75.8%; p = 0.03, 91.93% vs 74.19%; p = 0.008, respectively). CONCLUSION: In terms of lower postoperative pain and a better QOL within the early recovery period, LPSW is superior to LP alone. The saline washout procedure should be recommended during LC because it is a simple way to reduce pain intensity, even after LP operations. PMID- 15803239 TI - Thoracoscopic removal of neurogenic mediastinal tumors: technical aspects. AB - BACKGROUND: Thoracoscopy is fast becoming the standard approach for the removal of neurogenic mediastinal tumors. However, there are risks for adjacent nervous structures (stellate ganglion, spinal cord). The aim of this study was to review the technical features of this approach. METHODS: Between December 1999 and January 2003, nine patients underwent thoracoscopic resection of a mediastinal neurogenic tumor at our hospital. Five of these patients were asymptomatic with incidentally found tumor; the other four patients had compression-related syndromes. Two tumors had developed in the superior sulcus, and one had a spinal canal component (dumbell-type tumor). RESULTS: Thoracoscopic dissection was possible in all cases. In one patient, resection of the tumor was performed via a combined neurosurgical and thoracoscopic approach. Seven tumors were benign nerve sheath tumors (schwannoma), and 2 were nerve cell tumors (ganglioneuroma). The postoperative course was uncomplicated in all patients. CONCLUSION: The thoracoscopic resection of mediastinal neurogenic tumors is technically easy, except for bulky tumors of the superior sulcus and dumbbell tumors, which require a combined thoracoscopic and neurosurgical approach. PMID- 15803240 TI - The oxidative effect of prolonged CO2 pneumoperitoneum on renal tissue of rats. AB - BACKGROUND: Despite its advantages, laparoscopic donor nephrectomy is associated with prolonged operation time, which could potentially increase oxidative stress in the graft. We performed the first experimental, randomized, controlled study with blind assessment of outcome to address this possibility. METHODS: Wistar Albino rats were randomized into three groups. The animals in the control group were subjected to a sham operation under anesthesia; the animals in the other two groups were subjected to CO(2) pneumoperitoneum (Pp) for 120 and 240 min, respectively. The kidneys were removed at the end of each experiment. The concentrations of protein carbonyl and sulfhydryl (SH) groups and the activities of superoxide dismutase (SOD) were measured in renal tissue samples as markers of oxidative stress. Renal tissue samples were also evaluated histopathologically using light microscopy. RESULTS: Exposure to 120 min of Pp significantly increased the finding of oxidative stress in renal tissue samples, with an increase in protein carbonyl content and a decrease in protein sulfhydryls and tissue (SOD) activities. When exposure to Pp was prolonged from 120 min to 240 min, Pp associated oxidative stress was found to be increased. These changes occurred in the absence of light microscopical evidence of overt tissue damage. CONCLUSIONS: In an experimental model resembling laparoscopic donor nephrectomy, we found that exposure of pneumoperitoneum prolonged from 120 min to 240 min acts as an additive factor with respect to causing increased oxidative stress in renal tissue. Because these effects imply subtle tissue injury that may contribute to the chronic demise of renal grafts obtained laparoscopically, avoiding the use of Pp if possible and keeping operation time less than 120 min during laparoscopic donor nephrectomy appear to be advisable. PMID- 15803242 TI - Multimedia article: management of duodenal ulcer perforation with combined laparoscopic and endoscopic methods. AB - In the past, primary closes for perforated duodenal ulcer were not effective because of the high incidence of recurrence (42%), therefore most surgeons preferred definitive procedures to control acid production. Since the association of Helicobacter pylori with peptic ulcers and the effectiveness of the medication for acid control, definitive surgery is less frequently used. Today, primary closes of the perforation are the immediate alternative. Furthermore laparoscopic repair allows to perform the same technique than open surgery but with the benefits of the minimum invasion. The video shows the management of a 51-year-old woman who came to the emergency room with acute abdomen, leukocytosis, and fever. Laparoscopy showed peritonitis with apparent gastric perforation. Duodenal perforation was confirmed by endoscopy. Primary closes of the perforation, cavity cleaning, and placement of drainages were performed by laparoscopy. Evolution was excellent; the patient left the hospital 3 days after surgery. The contribution of laparoscopic and endoscopic surgery were shown to be highly beneficial for the patient, since an endoscopic view of the stomach and the duodenum was the pertinent approach to assure the correct perforation closes as well as to obtain diagnostic biopsy. PMID- 15803241 TI - Morphology of the rat peritoneum after carbon dioxide and helium pneumoperitoneum: a scanning electron microscopic study. AB - BACKGROUND: Laparoscopic surgery for patients with cancer has been debated because of the susceptibility that laparoscopic incisions have shown for metastatic tumor growth. Structural damage of the mesothelial layer attributable to the pneumoperitoneum may facilitate intraabdominal tumor cell adhesion and growth. The influence of carbon dioxide (CO(2)) and helium pneumoperitoneum on the morphology of the peritoneum was examined. METHODS: A total of 50 rats received colon carcinoma (DHB/TRb) cells intraperitoneally and CO(2) (n = 25) or helium (n = 25) pneumoperitoneum at 15 mmHg for 15 min. After different periods (2, 12, 24, 48, and 96 h), the rats were killed, and the peritoneum was examined by scanning electron microscopy. Control animals (n = 5) were without pneumoperitoneum. RESULTS: The control animals and most of the rats with pneumoperitoneum showed no peritoneal alterations. In four animals of each group, inflammatory alterations of the peritoneum such as bulging and retraction of mesothelial cells were observed at different time points. Tumor cells adherent to the peritoneum were found in a total of six animals. Peritoneal carcinomatosis, tumor nodules, or infiltration of the peritoneum by tumor cells was not observed. CONCLUSIONS: The study demonstrated that the morphologic integrity of the rat peritoneum is not disturbed when CO(2) or helium is used for insufflation combined with the intraperitoneal injection of carcinoma cells. Pneumoperitoneum therefore probably is not the condition causing peritoneal changes that favor intraperitoneal tumor growth. PMID- 15803243 TI - Videolaparoscopic treatment for isolated necrosis and abscess of the round ligament of the liver. AB - A case involving abcess and necrosis of the round ligament of the liver is described. This type of case is seldom reported in medical literature. Laparaoscopy is a very useful and feasible tool for the diagnosis and treatment of such cases. The video shows an oversized round ligament with necrotic appearance partially blocked by the epiplon, gallbladder, and stomach. (This online case report contains a video.). PMID- 15803244 TI - A robotic wide-angle view endoscope using wedge prisms. AB - BACKGROUND: The aim was to develop a new type of robotic wide-angle view endoscope that does not require rotation or bending of the endoscope to move the field of view. This endoscope allows the surgeon to make extensive observations in a small space. It is safe because it avoids the possibility of hitting organs accidentally. METHODS: The system consists of an endoscope with a zoom facility and two wedge prisms at the tip. Each prism is set on its own sleeve at the end of the endoscope, and both prisms rotate independently about the axis of the endoscope. RESULTS: In the in vivo experiment, the high quality of the image was confirmed, and almost whole liver could be observed without the observer moving the endoscope itself. CONCLUSIONS: This system provides safe robotic endoscopic surgery because of the high quality of the image. It has great significance for future clinical use. PMID- 15803246 TI - Noninvasive imaging of the common bile duct before invasive imaging after biliary pancreatitis. PMID- 15803249 TI - Sexuality before and after male-to-female sex reassignment surgery. AB - The sexual behaviors and attitudes of male-to-female (MtF) transsexuals have not been investigated systematically. This study presents information about sexuality before and after sex reassignment surgery (SRS), as reported by 232 MtF patients of one surgeon. Data were collected using self-administered questionnaires. The mean age of participants at time of SRS was 44 years (range, 18-70 years). Before SRS, 54% of participants had been predominantly attracted to women and 9% had been predominantly attracted to men. After SRS, these figures were 25% and 34%, respectively.Participants' median numbers of sexual partners before SRS and in the last 12 months after SRS were 6 and 1, respectively. Participants' reported number of sexual partners before SRS was similar to the number of partners reported by male participants in the National Health and Social Life Survey (NHSLS). After SRS, 32% of participants reported no sexual partners in the last 12 months, higher than reported by male or female participants in the NHSLS. Bisexual participants reported more partners before and after SRS than did other participants. 49% of participants reported hundreds of episodes or more of sexual arousal to cross-dressing or cross-gender fantasy (autogynephilia) before SRS; after SRS, only 3% so reported. More frequent autogynephilic arousal after SRS was correlated with more frequent masturbation, a larger number of sexual partners, and more frequent partnered sexual activity. 85% of participants experienced orgasm at least occasionally after SRS and 55% ejaculated with orgasm. PMID- 15803248 TI - Measurement of sexual arousal in postoperative male-to-female transsexuals using vaginal photoplethysmography. AB - Men's sexual arousal patterns are category-specific: Men typically display significantly greater physiological responses to sexual stimuli depicting members of their preferred gender category. Category-specific patterns of sexual arousal have not been consistently reported in natal women. We used vaginal photoplethysmography to examine patterns of sexual arousal in 11 male-to-female (MtF) transsexuals following sex reassignment surgery (SRS) and in 72 natal women. Subjective arousal was measured with a continuous response lever. Video clips depicting sexual activity between two males, two females, or one male and one female were used as erotic stimuli. All transsexual participants displayed category-specific sexual arousal. Five homosexual transsexual participants (attracted exclusively to males before sex reassignment) showed greater genital and subjective responses to male than to female stimuli, while six nonhomosexual transsexual participants showed the opposite pattern. Vaginal pulse amplitude (VPA) was lower in transsexual participants than in natal women. The mean correlation between VPA and subjective responses was high in nonhomosexual transsexuals, but was significantly lower in homosexual transsexuals and in natal women. One transsexual participant who reported a change in sexual orientation following sex reassignment displayed VPA and subjective responses consistent with her pre-reassignment sexual orientation. We conclude that male-to-female transsexuals display male-typical category-specific sexual arousal following SRS, and that vaginal photoplethysmography is a promising methodology for studying patterns of sexual arousal in postoperative transsexuals. PMID- 15803250 TI - Handedness, functional cerebral hemispheric lateralization, and cognition in male to-female transsexuals receiving cross-sex hormone treatment. AB - This study examined the impact of sex hormones on functional cerebral hemispheric lateralization and cognition in a group of male-to-female transsexuals receiving cross-sex hormone therapy compared to eugonadal men with a male gender identity. Cerebral lateralization was measured with a handedness questionnaire and a visual split-field paradigm and cognitive tests sensitive to sex hormone exposure (identical pictures, 3-D mental rotation, building memory) were also administered. Endocrine measures on the day of participation for transsexual and control subjects included total testosterone, free testosterone, estradiol, gonadotropins, and sex hormone binding globulin concentrations. Compared to controls, male-to-female transsexuals had elevated estradiol and sex hormone binding globulin concentrations and suppressed testosterone concentrations. Transsexual subjects showed a trend toward less exclusive right-handedness than controls. No group differences were observed on the visual-split-field or cognitive tasks. No direct associations were observed between endocrine measures and the laterality measures and cognitive performance. Previous observations of female-typical patterns in cerebral lateralization and cognitive performance in male-to-female transsexuals were not found in the current study. PMID- 15803251 TI - Sex differences in the flexibility of sexual orientation: a multidimensional retrospective assessment. AB - The flexibility of sexual orientation in men and women was examined by assessing self-reported change over time for three dimensions of sexual orientation (sexual fantasy, romantic attraction, and sexual behavior) across three categorical classifications of current sexual orientation (heterosexual, bisexual, and gay). The primary purpose of the study was to determine if there were sex differences in the flexibility (i.e., change over time) of sexual orientation and how such differences were manifested across different dimensions of orientation over the lifespan. Retrospective, life-long ratings of sexual orientation were made by 762 currently self-identified heterosexual, bisexual, and gay men and women, aged 36 to 60, via a self-report questionnaire. Cumulative change scores were derived for each of the three dimensions (fantasy, romantic attraction, and sexual behavior) of orientation by summing the differences between ratings over consecutive 5-year historical time periods (from age 16 to the present). Sex differences were observed for most, but not all, classification groups. There were significant sex differences in reported change in orientation over time for gays and heterosexuals, with women reporting greater change in orientation over time than did men. Bisexual men and women did not differ with respect to self-reported change in orientation. PMID- 15803252 TI - Same-sex sexual orientation, childhood sexual abuse, and suicidal behavior in university students in Turkey. AB - This questionnaire study investigated the prevalence of, and interrelationships among, self-reported same-sex sexual orientation, childhood sexual abuse, and suicidal behavior in 1262 university students in Turkey. Approximately 7% of the sample reported lifetime or current same-sex sexual attractions, 5% reported that they engaged in same-sex sexual behavior, and almost 2% self-identified as either homosexual or bisexual. Overall, almost 10% of the sample acknowledged some form of a same-sex sexual orientation. Twenty-eight percent of the participants reported at least one instance of sexual abuse during their childhood. Almost 42% of the students reported suicidal ideation during the past 12 months or lifetime, and 7% reported that they attempted to kill themselves during their lifetime or in the past 12 months. Five hypotheses about the interrelationships among same sex sexual orientation, childhood sexual abuse, and suicidal behavior were developed and tested in the study. Self-reported childhood sexual abuse was associated with same-sex sexual behavior. Participants who engaged in same-sex sexual behavior and those who identified themselves as homosexual or bisexual perceived more distance between themselves and their fathers than those who did not. Being sexually abused by someone of one's own sex was related to same-sex sexual orientation in male participants but not in female participants. Childhood sexual abuse was found to be an independent predictor of both suicidal ideation and attempts during the past 12 months. Only identifying oneself as homosexual or bisexual was associated with an increased risk for suicidal ideation during the past 12 months. PMID- 15803253 TI - Timing of puberty and sexuality in men and women. AB - We examined the relations among timing of puberty, timing of first experience of sexual arousal, strength of sex drive, and sexual attitudes and behaviors in 277 men (M age, 22.4 years) and women (M age, 21.8 years). Kinsey had suggested that earlier maturers have a stronger sex drive and, therefore, engage in a higher frequency of sexual behaviors (including same-sex contacts) than do later maturers. The purpose of this study was to test Kinsey's claim. Participants completed questionnaires on pubertal timing, timing of first experience of sexual arousal, sex drive in adulthood, sexual attitudes (e.g., sociosexuality or degree of comfort with casual sex), and sexual behaviors (e.g., lifetime number of sexual partners). Timing of puberty (i.e., emergence of secondary sex characteristics) was related to these variables for men but not for women. Timing of first sexual arousal was related to several aspects of adult sexuality in both sexes, but particularly in women. Earlier first sexual arousal was associated with having a higher sex drive, a less restricted sociosexual orientation, and with having had more sexual partners than was later first sexual arousal. Earlier first sexual arousal, but not timing of puberty, was related to sexual orientation for women only. We discuss classes of explanations for these results. PMID- 15803254 TI - Tactile sensitivity in women with sexual arousal disorder. AB - Evidence suggests that tactile sensitivity may differ between women with sexual arousal difficulties and women with normal sexual functioning. Tactile sensitivity was examined on the distal portion of the dominant hand index finger and on the lower lip in women with female sexual arousal disorder (FSAD) (n = 17) and in normally functioning women (n = 17). The two groups did not differ significantly in age, length of current relationship or on measures of sexual experience and sexual desire. Hierarchical binary logistic regression indicated that finger threshold was significantly associated with FSAD women versus control women, and hierarchical linear regression indicated that finger threshold was associated with severity of arousal dysfunction. Logistic regression showed that 76.5% of participants were correctly classified and 23.5% were incorrectly classified using tactile sensation as a predictor variable. Possible underlying mechanisms and clinical implications are discussed. PMID- 15803255 TI - Assessment of sexual behavior, sexual attitudes, and sexual risk in Sweden (1989 2003). AB - To assess changes in sexual behavior, sexual attitudes, and sexual risk related to HIV, we conducted mailed questionnaire surveys in random samples of the Swedish general population in 1989, 1994, 1997, 2000, and 2003 (total N = 13,762). Each sample consisted of 4,000-6,000 subjects aged 16-44 years, stratified by age: 16-17, 18-19, 20-24, 25-34, and 35-44 years. The overall participation rate was 63.8% (for men, 55.9%; for women, 71.9%). The prevalence of three or more sexual partners and casual sexual contacts without the use of a condom was comparatively high for men, for persons aged 16-24 years, single persons with and without a regular partner, and persons living in towns and urban areas. The prevalence of multiple sexual partners and casual sexual contacts increased significantly over time. There was a significant decrease in the proportion of participants who agreed with the statement "Sexual intercourse should only take place in a stable relationship." Personal risk assessments related to HIV did not change significantly over time. The study shows that risky sexual behavior related to HIV/AIDS increased in the Swedish population between 1989 and 2003, and that attitudes concerning casual sexual relations became more permissive. PMID- 15803256 TI - Sexual fantasies of adolescent male sex offenders in residential treatment: a descriptive study. AB - An attenuating sample of 87 male adolescent sexual offenders logged sexual fantasies during their entire time in therapy. These patients were attending residential treatment and kept a log recording all normal and deviant sexual fantasies which they experienced. Patients recorded which fantasies were interrupted and which fantasies were accompanied by masturbation. Typically, at the beginning of treatment, the reported rate of normal fantasies was maximal, the occurrence of deviant fantasies was minimal, and there was little effort to interrupt the deviant fantasies. The reported frequency of deviant fantasies increased substantially (about 380%) during the first 5 months of treatment and then steadily declined (approximately 47%) over the remaining months in therapy. Following an initial decline in the early months of treatment, an increase in the rates of normal fantasies was reported. Additionally, over the course of treatment, the frequency of deviant fantasies decreased relative to all fantasies. While deviant fantasies remained predominant to normal sexual fantasies and were more frequently paired with masturbation, patients reported substantial changes in the proportion of deviant fantasies that were interrupted (from about 20% to 70%). Implications of the results for juvenile sex offender treatment are discussed. PMID- 15803257 TI - Behavioral characteristics of men accused of rape: evidence for different types of rapists. AB - The aim of this study was to determine whether the behavioral characteristics demonstrated by rapists clustered together into groups that were similar to the common rapist typology in the literature: anger, power exploitative, power reassurance, and sadistic. Two studies were conducted to examine the evidence for this typology. Study 1 involved the analysis of data from 130 men charged with sexual assault and Study 2 involved the analysis of court transcripts from 50 rape cases tried through the court system. The results of Study 1 revealed that there was some validity to the characteristics usually associated with each of the four types of rape, especially for the power reassurance and sadistic rapists. However, there were some unexpected outliers within both the anger and power exploitative types of rapists, which may suggest that there is more than one type of anger rapist and more than two types of power rapists. The results of Study 2 very closely replicated the results of Study 1. Future research needs to focus on the behavioral, motivational, and cognitive characteristics associated with each of the types of rapists and research them separately, so that it is possible to further evaluate the evidence for the typology identified in this study. PMID- 15803258 TI - Heart failure and statins--why do we need a clinical trial? AB - The effect of statins to reduce mortality and morbidity in primary and secondary prevention as well as in acute coronary syndrome is well established. Recent data show that pleiotropic effects might also have direct effects on the myocardial cell. However, in chronic heart failure the outcome is inversely related to LDL plasma concentrations and other pleiotropic effects might impair mitochondrial function. Since there are no safety data on the use of statins in chronic heart failure, a controlled randomized and placebo-controlled trial is urgently needed. PMID- 15803259 TI - The relationship between hospital or operator volume and outcomes of coronary patients undergoing percutaneous coronary interventions. AB - The relationship between volume and outcome in medicine has been intensively investigated in the last few decades. The large amount of accumulated data demonstrates that for many surgical or non-surgical procedures and medical conditions, patients being treated in high-volume hospitals or by high-volume physicians have lower mortality rates and better quality of life compared to those treated by low-volume hospitals or by low-volume physicians. Although the degree of the relationship between high volume and better outcome varies, it is persistent across a wide range of procedures and conditions. Percutaneous coronary interventions (PCIs) have an important impact on public health, given the frequency of coronary heart disease for which these procedures are performed. Studies carried out before and after the advent of stents on the relationship between volume and outcome for PCIs have almost consistently reported that performance of PCIs in high-volume institutions or by high-volume operators is associated with improved outcomes for patients, regardless of the specific indication for PCI. For those procedures for which a relationship between high volume and better outcome has been clearly demonstrated, patients as well as their referring physicians should be informed that patients can benefit both in terms of reduced mortality and improved quality of life if they are treated by high-volume health care providers. Consequently, for these procedures, a health care policy aiming at their concentration in high-volume institutions should be strongly considered. PMID- 15803260 TI - Early time course of neointima formation and vascular remodelling following percutaneous coronary intervention and vascular brachytherapy of in-stent restenotic lesions as assessed by intravascular ultrasound analysis. AB - In-stent restenosis (ISR) represents the major limitation of stent implantation. Treatment, although of relative technical ease, is unsatisfactory due to a high incidence of recurrent restenosis. Vascular brachytherapy (VBT) has emerged as a powerful adjunct therapeutic modality to treat ISR. Inhibition of neointima formation has been regarded as the relevant mechanism of action. Yet, positive remodelling has been suspected as another contributing factor. Since only very few precise analyses of the extent, distribution and time course of the respective mechanims exist, the goal of the present study was to describe the changes of the vessel geometry at the target lesion and at the reference site following angioplasty and VBT of ISR in 42 patients by means of quantitative coronary angiography (QCA) and intravascular ultrasound (IVUS) before and after the index procedure and at the 3 and 6 month follow-up. By QCA the acute lumen gain measured 2.2+/-0.8 mm, the late lumen loss at 3 months was 0.1+/-0.5 mm and at 6 months 0.4+/-0.7 mm. By IVUS luminal cross-sectional area increased from 1.5+/-1.2 mm(2) to 7.9+/-1.9 mm(2) (p<0.001). The intima hyperplasia cross sectional area at 3 months was only 0.2+/-1.0 mm(2) (p=0.191), but increased to 0.7+/-0.6 mm(2) (p<0.001) at 6 months resulting in a lumen cross-sectional area of 7.1+/-1.7 mm(2). Stent dimensions did not show any significant changes over time. The external elastic membrane cross-sectional area at 3 months increased by 1.3+/-1.9 mm(2) (p<0.001), and showed a further increase by 0.7+/-2.9 mm(2) at 6 months. Positive remodelling could be demonstrated also at the reference segment. In conclusion the absolute amount of intima hyperplasia during a 6-month follow up period after VBT of ISR is low and most pronounced between the third and sixth month. Besides this, predominantly within the first 3 months of follow-up, significant positive remodelling could be demonstrated at the target lesion and at the reference site. Both observed effects may contribute to the preservation of the vessel lumen. PMID- 15803261 TI - N-Terminal-proBNP (NT-proBNP) as an indicator of cardiac dysfunction. A study in patients presenting with suspected cardiac disorders. AB - BACKGROUND: Natriuretic peptides represent a novel diagnostic tool in the assessment of cardiac dysfunction. METHODS: A total of 473 consecutive referred patients presenting to 18 cardiologists for the assessment of their cardiac state were recruited for the study. Patients received a medical history, a physical examination an electrocardiogram and an echocardiogram where left ventricular ejection fraction was recorded. RESULTS: NT-proBNP was found to correlate with left ventricular ejection fraction (LVEF), level of symptoms (NYHA classification), history of angina pectoris (AP) and myocardial infarction (AMI). Atrial fibrillation (AF) and thyroid dysfunction as well as renal impairment were shown to influence NT-proBNP levels. CONCLUSION: The study supports the hypothesis that NT-proBNP determination contributes to the assessment of patients presenting to cardiologists. PMID- 15803263 TI - Cardiac risk of coronary patients after reintegration into occupations with heavy physical exertion. AB - The job related reintegration of patients with coronary artery disease (CAD) is a central part of cardiac rehabilitation. However, specific occupational demands like jobs with heavy physical exertion (> 6 METs) could increase the cardiovascular risk because the relative risk for acute myocardial infarction (MI) and cardiac death is temporarily elevated after vigorous exertion ("hazard period"). Thus, in 2001 any male patient with proven CAD who performed a job with heavy exertion until the occurrence of an index event (MI/ACS, any interventional or surgical revascularization measure) received a questionnaire after an average of 20 months. Complete data were available in 108 from 119 included patients (90.8%), aged 51.8+/-7.8 years. Ejection fraction was 61.5+/-13.1% and the functional capacity at the time of hospital discharge averaged 130.1+/-31.2 W. 75% of the patients had a previous MI and 59.3% underwent bypass surgery. During follow-up the previous job with heavy exertion was performed over a cumulated time of 74 years. The aim of the study was to compare the observed and the expected incidence of MI and cardiac death with and without job performance. The expected ("basal") risk for MI and cardiac death without heavy physical exertion was determined from pooled study results and assumed to be 5.2% per year. The combined risk due to performing an occupation with strenuous exertion can be calculated from time periods with and without working hours and amounts to 11.9%. There could be expected 0.119 . 74=8.8 cardiac events related to the job. In contrast, 5 MIs (4 NSTEMI, 1 STEMI) were observed (6.8%). The relative risk for an expected event compared to the basal risk without heavy exertion was 2.3 (95% CI: 0.7-7.4). The relative risk for the observed cardiac events amounts to 1.3 (95% CI: 0.4-4.8). The lower observed risk is probably due to the high grade of physical fitness in this patient group. In spite of several limitations, our study showed no convincing evidence for increasing the cardiac risk of patients with CAD performing occupations with heavy physical exertion. Because of the importance of this prognostic finding, a representative and prospective study is strongly required. PMID- 15803262 TI - Peak oxygen uptake. Myth and truth about an internationally accepted reference value. AB - This article critically examines the execution of VO(2)-peak testing in cardiac patients and questions their appropriate interpretation. In the first part, the most common clinical implications of VO(2)peak measurements are discussed: assessment of (changes in) functional capacity, evaluation of the necessity of invasive diagnostic/therapeutic measures, reference for exercise prescriptions, determination of prognosis. In the second part, important methodological problems and constraints are addressed and illustrated by references to scientific studies. Finally, recommendations are given for meaningful VO(2)peak testing. It is evident that failure to strictly follow such recommendations might result in misleading ergometric findings and, thus, in over- or underestimation of endurance capacity and/ or training effects. PMID- 15803265 TI - Catheter ablation of junctional ectopic tachycardia in children, with preservation of atrioventricular conduction. AB - BACKGROUND: Idiopathic junctional ectopic tachycardia is a rare arrhythmia in children. Several studies have demonstrated that drug therapy is often ineffective and sometimes the only achieved effect is rate control. Early presentation and frequent recurrence are associated with adverse outcome. PATIENTS AND METHODS: Three consecutive children, aged 9, 7 and 12 years respectively, underwent radiofrequency catheter ablation for junctional ectopic tachycardia, after having failed antiarrhythmic drug therapy. The entire His bundle was plotted out and marked, using the Localisa navigation system. The arrhythmia was readily and repeatedly inducible using intravenous isoprenaline infusion and the site of earliest retrograde conduction during tachycardia could be assessed. Ablations were performed in sinus rhythm, empirically targeting the site of earliest retrograde conduction during tachycardia. RESULTS: This approach was successful in abolishing tachyarrhythmia in the first two patients, in whom the successful ablation site was located superoparaseptally. In the third patient, junctional ectopic tachycardia was inducible, despite abolishing retrograde atrial activation, in a septal location on the tricuspid valve annulus. Further ablations in the superoparaseptal region, closer to the His bundle, were successful in rendering tachyarrhythmia noninducible. Over a median follow-up of 10 months, none of the patients has had recurrence of arrhythmia, despite discontinuing all antiarrhythmic medications. CONCLUSIONS: Radio frequency catheter ablation of junctional ectopic tachycardia is feasible with preservation of atrioventricular conduction. PMID- 15803264 TI - [Acute coronary syndrome after diclofenac induced coronary spasm]. AB - We report about a 67-year old man, who was submitted to our clinic with acute coronary syndrome. The cardiac catheterization showed a proximal thrombus in the left anterior descending (LAD). The other coronary arteries did not have significant lesions. After percutaneous transluminal coronary angioplasty with stent-implantation into the proximal LAD the patient remained clinically stable. Cardiac enzymes confirmed no myocardial necrosis. Three days after the acute coronary syndrome the patient developed a podagra, which was treated with colchicinum, diclofenac and local cooling. Five hours after initial therapy the patient developed severe symptoms of angina pectoris and electrocardiographical signs of an acute posterior and anterior myocardial infarction. Immediate coronary angiography demonstrated extended vasospasm of the right coronary artery. Intracoronary application of verapamil and nitroglycerin resolved the coronary spasm. The patient reported about a self-indicated application of diclofenac six hours before hospital admission. This case demonstrates that oral application of diclofenac can provoke coronary vasospasm. PMID- 15803266 TI - [Position paper on "automated external defibrillation" ]. PMID- 15803271 TI - Bacterial adhesion to bisphosphonate coated hydroxyapatite. AB - Staphylococcus aureus (S. aureus) is commonly associated with microbial infection of orthopaedic implants. Such infections often lead to osteomyelitis, which may result in failure of the implant due to localised bone destruction. Bacterial adhesion and subsequent colonisation of the device may occur as a consequence of contamination during surgery, or by seeding from a distant site through the blood circulation. Coating of the hydroxyapatite (HA) ceramic component of artificial hip joints with the bisphosphonates clodronate (C) and pamidronate (P) has been proposed as a means to minimise osteolysis and thereby prevent loosening of the implant. However, the effect of the bisphosphonate coating on bacterial adhesion to the HA materials must be determined before this approach can be implemented. In this study coated HA materials were incubated with the S. aureus and the number of adherent bacteria determined using the Modified Vortex Device (MVD) method. The number of bacteria adherent to the P coated HA material was significantly greater than that adherent to uncoated HA (60-fold increase) or to the C coated HA (90-fold increase). Therefore, even though earlier studies suggested that P bound to HA may improve osseointegration, the results presented would suggest that the use of this coating may be limited by the potential increased susceptibility of the coated device to infection. PMID- 15803272 TI - Polymeric meshes for internal sutures with differentiated adhesion on the two sides. AB - The aim of this work is to investigate the effects of different plasma treatments on ePTFE abdominal prostheses with the final goal of obtaining a new prosthesis, made of a single strand of ePTFE, with clearly differentiated adhesion properties on the two sides, which should be able to promote tissue ingrowth on one side and prevent post surgical visceral adhesions on the other. Samples obtained from ePTFE Bard Dulex Meshes have been treated sequentially with three different gases (N(2), O(2) and NH(3)) in order to choose the optimal treatment conditions to improve ePTFE wettability. In particular, no modification was induced by N(2) treatment, while the full treatment after the final ammonia gas resulted in the best suitable candidate. As demonstrated by scanning electron microscopy, AFM analyses and contact angle measurements, ammonia plasma treatment increases ePTFE surface roughness and renders it more hydrophilic, thus promoting adhesion without any alteration of the material's bulk properties. The reported results also evidence the possibility to obtain the maximum wettability with a cheap treatment by optimizing plasma exposure time. As a preliminary cell adhesion study, Swiss 3T3 fibroblasts (mouse, embryo) have been seeded on the treated and untreated materials in order to assess whether there was any difference in terms of cell attachment and spreading. Cells seeded on the ammonia plasma treated material showed a better adhesion and spreading when compared to the untreated material. PMID- 15803273 TI - Salivary bisphenol-A levels detected by ELISA after restoration with composite resin. AB - Bisphenol-A diglycidylether methacrylate (Bis-GMA), which is synthesized from bisphenol-A (BPA), a compound with exogenous endocrine disrupter action, is widely used as a dental material. During clinical filling with sealants and composite resins, the compounds are solidified by polymerization and then used. However, it has been noted that unpolymerized monomers may become dissolved in saliva. In this study using a competitive ELISA system, we investigated the changes in the BPA concentration in saliva after restoration with composite resins. Commercial composite resins from nine companies were tested. Mixed saliva was collected from 21 subjects. Based on the dynamics of salivary BPA detected by this ELISA system, we concluded that several tens to 100 ng/ml of BPA were contained in saliva after filling teeth with composite resin but that sufficient gargling can remove it from the oral cavity. Our data suggest that sufficient gargling after treatment is important for risk management. PMID- 15803274 TI - Characterization of surface modified Ti-6Al-7Nb alloy. AB - In the last years different types of surface modifications were developed with the aim of improving the osteointegration ability of titanium alloys. The chemical composition, crystallographic structure and morphology of a surface layer can be modified in order to obtain a better interaction between the implant, the cells and the organic fluids. The final goal is to obtain a more efficient bone growth also in critical clinical cases. In the present paper were reported several data about the characterization of the Ti-6Al-7Nb alloy treated by two innovative surface treatments. They consist of blasting, followed by a two step chemical etching and heat treatment performed in air or in vacuum. TEM, XRD and SEM investigations were performed in order to assess the structure and morphology of the modified surfaces. The surface chemical composition was investigated by XPS ad AES analyses. The ability to interact with physiological fluids was tested by immersion of the treated materials in an acellular simulated body fluid (SBF). Metal ion concentration analyses of the fluid and SEM observations of the samples were performed after different times of soaking. The mechanical characterization involved scratch and fatigue tests. The surface of treated samples shows chemical, structural and morphological modifications. The passivation pre-treatment has influence on the surface modification. The treated samples evidenced a quite low metal ion release and interact with SBF solution, showing a moderate bioactivity. A relevant decrease in fatigue strength was observed on modified samples. PMID- 15803275 TI - Effects of bacterial adhesion with respect to the type of material, structure and design of intraocular lenses. AB - The properties of the biomaterials used to constitute lenses are important factors choosing a lens for human implantation because these can influence in posterior clinical evolutions of patients. In this study, different characteristics of intraocular lenses such as chemical composition, surface roughness and lens design have been investigated in terms of their influence into a pathological environment. Eight commercial lenses were tested by optical profiling, Infrared spectra with Fourier transformation (FTIR), water-material contact angle and scanning electron microscope (SEM) to know their chemical composition and structural characteristics. These lenses were then exposed to infectious conditions in order to evaluate their responses to the bacterial environment. PMID- 15803276 TI - Precipitation of hydroxyapatite nanoparticles: effects of precipitation method on electrophoretic deposition. AB - Electrophoretic deposition is a low-cost, simple, and flexible coating method for producing hydroxyapatite (HA) coatings on metal implants with a broad range of thicknesses, from < 1 microm to > 500 microm. As for many other HA coating techniques, densification of electrophoretically deposited coatings involves heating the coated metal to temperatures above 1000 degrees C. Metal substrates tend to react with HA coatings at such temperatures inducing decomposition at temperatures below 1050 degrees C (decomposition for pure HA normally occurs above 1300 degrees C). Therefore, densification of these coatings needs to be conducted at temperatures lower than 1050 degrees C, and this necessitates the use of high-surface-area HA nano-precipitates, rather than commercially available pre-calcined powders, which densify at temperatures typically higher than 1200 degrees C. HA nano-precipitates were prepared by three methods and deposited on metal substrates by electrophoresis: (1) the acid base method, which produced plate-like nano-particles with a 2.5:1 aspect ratio, and severely cracked coatings; (2) the calcium acetate method, which produced needle-like nano particles with a 10:1 aspect ratio, and slightly cracked coatings; (3) the metathesis method, which produced rounded nano-particles with a 2:1 aspect ratio, and high-quality crack-free coatings. The results suggested that the less equiaxed the nano-particles, the more cracked the coatings obtained by the electrophoretic deposition technique. PMID- 15803277 TI - Coralline hydroxyapatite reinforced with polylactide fibres in lumbar interbody implantation. AB - Porous hydroxyapatite HA blocks reinforced with poly-l/dl-lactide fibres were used to maintain the lumbar disc space and to start to create intercorporeal fusion in 23 growing pigs. In four pigs two emptied non adjacent disc spaces were left open. After 3, 6, 12 and 16 weeks the implanted disc blocks were studied radiologically, histologically, histomorphometrically, microradiographically, and with oxytetracycline fluorescence. In plain films slight to moderate ossification of the implanted disc spaces was detected at 12 and 16 weeks. Resorption of the implants was seen radiologically from 3 weeks and fragmentation from 12 weeks onwards. In microradiographs disintegration of the coralline inner structure started at 3 weeks. Histologically, connective tissue ingrowth was seen inside the porous structure from three weeks onwards. Small amounts of new bone were visible and connective tissue inside the implant increased from a mean of 65.6% at 3 weeks to a mean of 79.4% at 16 weeks histomorphometrically. The bone ingrowth varied from 0.7 to 1.7%. A loss of height in the implanted disc spaces was seen (p < 0.05, linear regression analysis). In control pigs the emptied disc spaces lost their height similarly. The implants used were not strong enough to maintain the lumbar disc height. PMID- 15803278 TI - Biomaterial with chemically engineered surface for protein immobilization. AB - The last 3 decades have been a revolution in the area of sol-gel-derived materials. They can be used to encapsulate biomolecules such as enzymes, antibodies, hormones, and proteins retaining their functional state. Proteins can be immobilized in many ways but it is crucial that they retain their native conformational structure and, therefore, bioactivity. Porous silica gel matrixes with modified surfaces offer unlimited possibilities to control the protein-solid interaction behavior. The bioimmobilization process on sol-gel biomaterials with chemically engineered surface has driven applications on solid-phase materials, affinity chromatography, biosensors and many others. In the present work, we have aimed to produce surface-modified silica glass materials obtained via sol-gel route to be used as solid support on drug delivery systems and as solid-phase in immunodiagnostic. The functionalization process was carried out by reacting alkoxysilanes with 5 different silane surface modifying chemical groups: tetraethoxysilane (TEOS), 3-mercaptopropyltrimethoxysilane (MPTMS) and 3 aminopropyltriethoxysilane (APTES), 3-glycidoxypropyltrimethoxysilane (GPTMS) and 3-isocyanatopropyltriethoxysilane (ICPES). The bioactivity assays were based on two main tests: (a) An in vivo bioresponse of rats with sol-gel disk implants with insulin protein incorporated. In vivo tests with adult male rats were used to verify the immobilized insulin bioactivity after implantation of different biomaterial with functionalized surfaces. All surface modified materials have presented hypoglycemic peak response associated with the insulin bioactivity. (b) The produced solid-phase sol-gel disks with protein substrates were tested through Enzyme Linked Immuno Sorbent Assay (ELISA). The immunoassay results have showed that glasses with chemically functionalized surfaces regulated the extent of bioimmobilization of protein. The amine, thiol and hydroxyl terminated porous gels have showed significant interaction with the antibody-antigen, during the coupling process. We believe that it is due to balance of forces associated with Van der Waals interaction, hydrophilic and hydrophobic forces and steric hindrance acting at the surface. Therefore, such novel biomaterial could be advantageously used in drug delivery systems and in immunoassays of diagnostic kits. PMID- 15803279 TI - An in vivo study of a bone grafting material consisting of hydroxyapatite and reconstituted collagen. AB - This study aims to evaluate the performance of our recently developed microspheres of hydroxyapatite/reconstituted collagen as a bone grafting material. The microspheres were fabricated into a circular disc and implanted in a pre-drilled hole in a rat's calvaria. The bone tissue had regenerated and grown into the disc bone graft 4 weeks following implantation. After 16 weeks of implantation, the regenerated bone had integrated with the remaining material and made close contact with it. The disc had been completely absorbed with almost no visible bone graft left after 24 weeks of implantation. In contrast, a hydroxyapatite disc still remained intact on the 24th week after implantation. These results suggested that the hydroxyapatite/reconstituted collagen microsphere can be used as an excellent bone grafting material. PMID- 15803280 TI - Changes in surface characteristics of dental resin composites after polishing. AB - The objectives of this study were (1) to determine in vitro changes in surface roughness and color of dental resin composites after application of three finishing and polishing systems; (2) to evaluate the difference in color stability after immersion in a dye solution after polishing; and (3) to evaluate the effects of surface condition, especially roughness, on measured color depending on the color measuring geometries of specular component excluded (SCE) and specular component included (SCI). Color and surface roughness (R(a)) of resin composites of four brands of A2 shade and one brand of Yellow Enamel shade were measured after polymerization, after polishing with Enhance (Dentsply), Sof Lex (3M ESPE), or Super-Snap (Shofu) composite finishing and polishing systems. Color was also measured after immersion in 2% methylene blue solution. Color was measured according to the CIELAB color scale. Color changes (DeltaE*(ab)) after polishing/staining and by the measuring geometry were calculated by the equation; DeltaE*(ab) = [(DeltaL*)(2) + (Deltaa*)(2) + (Deltab*)(2)](1/2). Ra value was measured with a surface roughness tester. DeltaE*(ab) and DeltaL* values after polishing and after staining varied among polishing systems when measured with SCE geometry. Composites polished with Super-Snap and Sof-Lex systems showed higher DeltaE*(ab) and DeltaL* values than those polished with Enhance polishing system with SCE geometry. DeltaE*(ab) and DeltaL* values between specimens with different surface conditions measured with SCE geometry were significantly higher than those with SCI (p < 0.01). Changes in R(a) value after polishing was insignificant in most cases. PMID- 15803281 TI - Effects of silica on the bioactivity of calcium phosphate composites in vitro. AB - In the present study, silica-calcium phosphate composites (SiO(2)-CaP composites) were developed by mixing the starting materials (SiO(2) and CaHPO(4)) in different ratios with the addition of 0.1% w/v NaOH solution. The phase composition of the SiO(2)-CaP composites was determined by XRD and FTIR. After thermal treatment at 350 degrees C/1 h and at 1000 degrees C/3.5 h; all SiO(2) CaP composites composed of beta-quartz, alpha-cristobalite and beta-Ca2P2O7. The presence of calcium phosphate enhanced the transformation of beta-quartz into alpha-cristobalite at 1000 degrees C. SEM observation indicated favorable attachment and spreading of neonatal rat calvaria osteoblasts onto the surface of silica-rich SiO(2)-CaP composites. After attachment, these cells produced significantly higher amount of protein and expressed higher AP activity than cells attached to silica-poor samples. Results of the study suggested that the silica-based composites are more bioactive than calcium phosphate-based composites. Silica promoted the expression of osteoblast phenotype by both solution-mediated effect and direct interaction with the surface of the substrate. PMID- 15803282 TI - Preparation and characterization of bioactive collagen/wollastonite composite scaffolds. AB - A novel biodegradable collagen/wollastonite composite was prepared as three dimensional scaffolds by freeze-drying method. Scanning electron microscope (SEM) micrographs of scaffolds showed a continuous structure of interconnected pores, and pore size was about 100 microm. The tensile strength of the scaffolds was improved by incorporation of wollastonite and the in vitro bioactivity of the scaffolds was evaluated by examining the hydroxyapatite (HA) deposition on their surface in simulated body fluid (SBF). After soaking in SBF for 7 days, collagen reconstituted to fibers and HA nodules formed on collagen fibers. The result suggests that the incorporation of wollastonite could improve the mechanical strength and the in vitro bioactivity of the composite. The scaffolds could be a potential biomaterial for bone tissue engineering. PMID- 15803284 TI - Abstracts of the 6th International Symposium on Osteoporosis: Current Status and Future Directions, 6-10 April 2005, Washington, DC, USA. PMID- 15803283 TI - Development of fibrous biodegradable polymer conduits for guided nerve regeneration. AB - The technique of microbraiding with modification was employed as a novel method for the fabrication of fibrous tubular scaffolds for nerve tissue engineering purposes. The biodegradable polymers used in this study were poly(L-lactide-co glycolide) (10:90) and chitosan. The polymeric fibers were microbraided around a Teflon mandrel to make it as a tubular construct. The conduits were then studied for their surface morphology, swelling behaviour and biocompatibility. The surface morphology was analysed by scanning electron microscope, swelling behaviour by weight increase due to water uptake and biocompatibility by in vitro cytotoxicity assessment in terms of cell morphology and cell viability by the MTT assay of polymer extract treated cells. These conduits may also be used for regeneration of tissues, which require tubular scaffolds such as blood vessel, spinal cord, intestine etc. PMID- 15803286 TI - In vivo study of the healing processes that occur in the jaws of rabbits following perforation by an Er,Cr:YSGG laser. AB - This study investigates the healing process that takes place in the bone and soft tissue of the maxilla and the mandible after perforation by an Er,Cr:YSGG laser device. The jaws of New Zealand white rabbits were irradiated with an Er,Cr:YSGG laser, forming wounds 0.4 mm in diameter. Irradiation parameters were as follows: repetition rate was 20 pulse/s, pulse duration was 140-200 micros, power was 2 W, exposure time was 10 s, energy density was 80 J/cm(2). After sacrifice at 0-56 days post-surgery, gross observations and histological examinations were performed. Effective hemostasis was achieved after Er,Cr:YSGG laser surgery. There was a minimal delay before the healing began. After 56 days all of the bone defects had been completely replaced by new bone. In conclusion, the Er,Cr:YSGG laser allows precise surgical ablation with minimal thermal damage to adjacent tissues in vivo. The overall subsequent healing was favorable. This laser may potentially be used in minor oral surgery. PMID- 15803285 TI - Relations between pregnancy-related low back pain, pelvic floor activity and pelvic floor dysfunction. AB - To assess the occurrence of pelvic floor dysfunction (PFD) in pregnancy- related low back and pelvic pain (PLBP) patients, a cross-sectional study was performed, comprising 77 subjects. Each subject underwent physical assessment, and filled in the Urogenital Distress Inventory completed with gynaecological questions. Differences in the presence of PFD between PLBP patients and healthy controls as well as differences in pelvic floor muscle activity were tested for significance. Interaction by age and vaginal delivery were tested. PFD occurred in 52% of all PLBP patients, significantly more than in the healthy control group. In PLBP patients a significantly increased activity of the pelvic floor muscles could be demonstrated with respect to healthy controls. The occurrence of PFD and PLBP was influenced by a confounding effect of age. Clinicians should be aware of the relation between PLBP and PFD and hence address both problems at the same time. PMID- 15803287 TI - [The effect of combined mitoxantrone and methylprednisolone therapy in primary and secondary progressive multiple sclerosis. An applied study in 65 patients]. AB - Mitoxantrone (mitox) has been shown to be effective for secondary progressive (SP) and relapsing-remitting multiple sclerosis (MS). The aim of this open trial was to evaluate the effects of combined mitox and methylprednisolone (MP) therapy on patients with primary progressive (PP)-MS or with SP-MS. We present here the results of an interim analysis done after the study had lasted 5 years. Sixty five patients (20 with PP-MS and 45 with SP-MS) have been included so far. The treatment involved ten cycles of combined mitox and MP. The intervals between the individual cycles were systematically prolonged from 3 months initially to 12 months, so the complete treatment took a total of 57 months. Conclusion This interim analysis indicates that mitox combined with MP beneficially reduces the progression of disability in patients with PP-MS and SP-MS. Therefore, this therapy regimen can also be considered a feasible option for PP-MS. PMID- 15803288 TI - H9, H10, and H11 compose a cluster of Hessian fly-resistance genes in the distal gene-rich region of wheat chromosome 1AS. AB - H9, H10, and H11 are major dominant resistance genes in wheat, expressing antibiosis against Hessian fly [(Hf) Mayetiola destructor (Say)] larvae. Previously, H9 and H10 were assigned to chromosome 5A and H11 to 1A. The objectives of this study were to identify simple-sequence-repeat (SSR) markers for fine mapping of these genes and for marker-assisted selection in wheat breeding. Contrary to previous results, H9 and H10 did not show linkage with SSR markers on chromosome 5A. Instead, H9, H10, and H11 are linked with SSR markers on the short arm of chromosome 1A. Both H9 and H10 are tightly linked to flanking markers Xbarc263 and Xcfa2153 within a genetic distance of 0.3-0.5 cM. H11 is tightly linked to flanking markers Xcfa2153 and Xbarc263 at genetic distances of 0.3 cM and 1.7 cM. Deletion bin mapping assigned these markers and genes to the distal 14% of chromosome arm 1AS, where another Hf-resistance gene, Hdic (derived from emmer wheat), was also mapped previously. Marker polymorphism results indicated that a small terminal segment of chromosome 1AS containing H9 or H10 was transferred from the donor parent to the wheat lines Iris or Joy, and a small intercalary fragment carrying H11 was transferred from the resistant donor to the wheat line Karen. Our results suggest that H9, H10, H11, Hdic, and the previously identified H9- or H11-linked genes (H3, H5, H6, H12, H14, H15, H16, H17, H19, H28, and H29) may compose a cluster (or family) of Hf-resistance genes in the distal gene-rich region of wheat chromosome 1AS; and H10 most likely is the same gene as H9. PMID- 15803289 TI - Locating the broad-spectrum wheat leaf rust resistance gene Lr52 (LrW) to chromosome 5B by a new cytogenetic method. AB - This study was conducted to genetically map a potentially new wheat leaf rust resistance gene (LrW) using a novel genetic method and to test its effectiveness against current races of leaf rust (Puccinia triticina Eriks.) in Canada. Undoubled haploids of a near-isogenic line of Thatcher carrying the resistance gene (RL6107) were pollinated with a contrasting susceptible cultivar to generate an array of hybrids with random deficiencies arising from irregular meiosis of the haploid. Genetic analysis of the deficiencies in such populations can be used to locate qualitative traits by which the two parents differ through a process that we have called haploid deficiency mapping. In the present case, 5/417 hybrids were both susceptible to leaf rust (i.e. lacked the resistance gene) and also lacked several polymorphic microsatellite alleles from RL6107 that are specific to chromosome 5B. This correlated failed transmission of the resistance gene and deficiency for chromosome 5B. Analysis of an F2 population showed that the factor conditioning resistance was located on the short arm of 5B, 16.5 cM distal to the locus of the microsatellite Xgwm443. Since no other leaf rust resistance genes have been mapped to this region, LrW was re-designated Lr52. RL6107 was tested with 29 isolates of P. triticina, encompassing a diversity of virulence found in North America, with none showing virulence. The effectiveness and novelty of Lr52 make it a promising source of resistance for North American wheat cultivars. PMID- 15803290 TI - Mapping QTLs for grain dormancy on wheat chromosome 3A and the group 4 chromosomes, and their combined effect. AB - A major QTL for grain dormancy, QPhs.ocs-3A.1, derived from the highly dormant wheat Zenkoujikomugi (Zen), has been identified in a study made under a controlled environment. Further investigations were needed to dissect the precise position and expression of QPhs.ocs-3A.1 under different field conditions because the ability to detect genetic loci for grain dormancy traits is compromised by environmental effects and genotype/environment interactions. Group 4 chromosomes have also been shown to be possible sites of QTLs for grain dormancy. The objectives of this study were (1) to locate additional molecular markers in the QPhs.ocs-3A.1 region, (2) to identify QTLs on the group 4 chromosomes and (3) to elucidate their combined effects. We examined the recombinant inbred lines (RILs) from a cross between Chinese Spring (CS) and Zen over a 3-year period in one location and 1 year in a different location. In an interval mapping study QPhs.ocs-3A.1 was mapped to within the 4.6 cM region flanked by Xbarc310 and Xbcd907 at the proximal end of the short arm of chromosome 3A. QPhs.ocs-3A.1 was confirmed to be the predominant dormancy QTL since it explained a large portion (11.6-44.8%) of the phenotypic variation, and was strongly displayed under dormancy-breaking conditions or at low germination temperatures. For QPhs.ocs 4A.1, identified on the long arm of chromosome 4A, and QPhs.ocs-4B.1, on the centromeric region of the long arm of Chr 4B, the LOD peak positions and the desirable allele were consistent between the trials, while the LOD scores and contribution to the phenotypic variation varied. Transgressive segregants were observed among the 125 RILs and most of them had a combination of the three alleles conferring a higher dormancy: the Zen alleles at QPhs.ocs-3A.1 and QPhs.ocs-4A.1 and the CS allele at QPhs.ocs-4B1. This demonstrated a combined effect of the desirable alleles on accelerating grain dormancy, with their total effect being superior to that of Zen. PMID- 15803291 TI - Genetic diversity among barley cultivars assessed by sequence-specific amplification polymorphism. AB - We analyzed the genetic structure and relationships among barley cultivars (Hordeum vulgare L.) with sequence-specific amplification polymorphisms (S-SAPs). Polymorphisms were identified in 824 individual barley plants representing 103 cultivars (eight plants per cultivar) widely grown in Canada and the United States, using PCR primers designed from the long terminal repeat of the barley retrotransposon BARE-1 and a subset of four selective MseI primers. From the 404 bands scored, 150 were polymorphic either within or between cultivars. Genetic structure assessed with analysis of molecular variance attributed the largest component of variation to the within groups of cultivars (69-86%). Within cultivar genetic variation was estimated as average gene diversity over loci and ranged from 0 (completely homogenous) to 0.076 (most heterogeneous cultivar). Only 17 out of 103 cultivars (16%) were judged to be homogenous by this criterion. Relationships among cultivars were analyzed by cluster analysis using unweighted pair-groups using arithmetic averages and found groups similar to those determined by agriculturally significant phenotypic traits such as spike morphology (two-rowed or six-rowed), cultivar type (malting or feed), seed characteristic (hull-less or hulled), and growth habit (winter or spring), with minor overlaps. Discriminant analysis of groups determined by these phenotypic traits fully supported the different groups with minor overlaps between the malting/feed. S-SAP markers generated from retrotransposons such as BARE-1 are invaluable tools for the study of genetic diversity in organisms with a narrow genetic base such as barley. In this study, S-SAP analysis revealed significant amounts of cryptic variation in closely related cultivars including somaclonal variation, which could not be inferred by the pedigree analysis. PMID- 15803292 TI - On-going on-farm microevolutionary processes in neighbouring cowpea landraces revealed by molecular markers. AB - A knowledge of existing levels of diversity is fundamental for planning in situ (on-farm) conservation activities. Three neighbouring cowpea landraces (LRs) currently cultivated in central Italy were studied by amplified fragment length polymorphism (AFLP) and selectively amplified microsatellite polymorphic locus (SAMPL) markers to determine the distribution of genetic variation within and among them. The three LRs studied, although relatively similar, are highly different from one another as shown by the significance of the Fisher exact test for the genic differentiation and the absence of genotype sharing among them. Data obtained from the AFLP and SAMPL markers separately and their combined data revealed a relatively high level of diversity still present within the LRs. The more efficient SAMPL technique was better at discriminating between the plants than the AFLP markers. The three LRs studied appear to be structured as a metapopulation in which a substantial differentiation is maintained at the subpopulation level. A complex interaction of factors (drift, LR isolation, farmer selection, migration within LRs) explains the observed pattern of diversity. The results suggest that the best strategy for maintaining diversity in the area is to preserve each of the LRs observed on the farms from which it came. PMID- 15803293 TI - Extending the limits of extracorporeal membrane oxygenation: lung rest for a child with non-specific interstitial pneumonia. AB - OBJECTIVE: Veno-venous extracorporeal membrane oxygenation (ECMO) is an established therapy for the treatment of respiratory failure. Traditionally ECMO has been used to support patients with an acute, reversible disease process, with a predictable outcome. We report the successful use of veno-venous ECMO for an unusual indication. PATIENT: A 10-year old girl was admitted to intensive care with severe, hypoxic respiratory failure on the background of a 2-month history of worsening respiratory symptoms. She required high levels of conventional positive pressure ventilation, and high frequency oscillation. Lung biopsy confirmed a non-specific interstitial pneumonia, and the patient was commenced on immune suppressive therapy. Her clinical course was further complicated by pulmonary haemorrhage and severe air leak. INTERVENTIONS: On day 20 after admission the patient was placed on veno-venous ECMO for lung rest while awaiting a response to continued medical treatment. She required ECMO for 20 days, during which time sedation was reduced, and she was able to interact with those around her. The patient's ventilatory requirements after decannulation were minimal, and she subsequently made a steady clinical recovery. CONCLUSIONS: ECMO was safely and successfully used to provide a period of lung rest and time for medical therapy to take effect in a child with an unusual indication for support: a rare disease with an uncertain outcome on the background of prolonged mechanical ventilation. PMID- 15803294 TI - Comparison between cerebral tissue oxygenation index measured by near-infrared spectroscopy and venous jugular bulb saturation in children. AB - OBJECTIVE: To compare the cerebral tissue oxygenation index (TOI) measured by near-infrared spectroscopy (NIRS) with venous oxygen saturation in the jugular bulb (SjO(2)) during elective cardiac catheterization in children. DESIGN AND SETTING: Prospective observational clinical study in a catheterization laboratory for pediatric cardiology. PATIENTS: Sixty children with congenital heart defects admitted to the catheterization laboratory. MEASUREMENTS AND RESULTS: TOI measured noninvasively by NIRS was compared to SjO(2) measured in the jugular bulb during cardiac catheterization. Patients were divided into two groups regarding body weight: below (n=29) and above 10 kg (n=31). Linear regression analysis and Pearson's correlation coefficient were calculated. Bland-Altman analysis, sensitivity, and specificity calculation for spatially resolved near infrared spectroscopy with a cutoff level of 60% were performed. Simultaneously measured values for SjO(2) (67.3+/-9.8%, 40-84.1%) and TOI (65.7+/-7.2%, 39-80%) showed a significant correlation; the correlation in children weighing under 10 kg was stronger in children weighing over 10 kg. Bland-Altman analysis showed a mean bias of -1.8% with limits of agreement between 11.7% and -15.3% for all children. Sensitivity and specificity of the SRS method were 46% and 91%, respectively, for all children and 53% and 83% respectively in infants weighing under 10 kg. CONCLUSIONS: The results demonstrate that despite a significant correlation, sensitivity of spatially resolved spectroscopy is poor, and it is questionable whether TOI can be used reliably to detect low SjO(2). PMID- 15803295 TI - Concurrent bacterial infection and prolonged mechanical ventilation in infants with respiratory syncytial virus lower respiratory tract disease. AB - OBJECTIVE: To identify demographic, clinical, and laboratory variables predictive for a concurrent bacterial pulmonary infection in ventilated infants with respiratory syncytial virus (RSV) lower respiratory tract disease (LRTD) and investigate antimicrobial drug use. DESIGN AND SETTING: Retrospective, observational study in a 14-bed pediatric intensive care unit. PATIENTS: 82 infants younger than 1 year of age with a virologically confirmed RSV LRTD during 1996-2001, of whom 65 were mechanically ventilated. RESULTS: Microbiological data were available from 38 ventilated infants, 10 of whom had a positive blood culture (n=1) or endotracheal aspirate (n=9) obtained upon admission to the pediatric intensive care unit (PICU). Infants with a positive culture had a lower mean gestational age but were otherwise demographically comparable to those with negative culture results. Infants with a positive culture were ventilated 4 days longer. Indicators for a concurrent bacterial infection were comparable between ventilated and nonventilated infants. Antimicrobial drugs were used in 95.1% of infants (100% of ventilated infants) with a mean duration of 7.8+/-0.3 days. The moment of initiation and duration of antimicrobial drug treatment varied considerably. CONCLUSIONS: We observed in ventilated infants a low occurrence of concurrent bacterial pulmonary infection, but infants with positive cultures needed prolonged ventilatory support. Improvement in the diagnosis of a pulmonary bacterial infection is warranted to reduce the overuse of antimicrobial drugs among ventilated infants with RSV LRTD and to restrict these drugs to the proper patients. PMID- 15803296 TI - The involvement of intensive care nurses in end-of-life decisions: a nationwide survey. AB - OBJECTIVE: To investigate the prevalence and predictors of intensive care nurses' active involvement in end-of-life (EOL) decisions. DESIGN AND SETTING: A survey of intensive care nurses from 36 intensive care units (ICUs) in New Zealand. MEASUREMENTS AND RESULTS: A total of 611 ICU nurses from 35 ICUs responded to this survey. The response rate was estimated to be between 43% and 81%. Seventy eight percent of respondents reported active involvement in EOL decisions, especially the senior nurses (level IV vs. I nurses, OR 7.9; nurse educators vs. level I nurses, OR 4.3). Asian (OR 0.2) and Pacific Islander nurses (OR 0.2) were less often involved than European nurses. Sixty-eight percent of respondents preferred more involvement in EOL decisions, and this preference was associated with the perception that EOL decisions are often made too late (OR 2.2). Sixty five percent believed their active involvement in EOL decisions would improve nursing job satisfaction. CONCLUSIONS: Most ICU nurses in New Zealand reported that they are often involved in EOL decisions, especially senior and European nurses. PMID- 15803298 TI - Congestive cardiomyopathy after streptococcal toxic shocklike syndrome. PMID- 15803297 TI - Infections in the survivors of out-of-hospital cardiac arrest in the first 7 days. AB - OBJECTIVE: To evaluate the incidence, risk factors, foci, isolated organisms, and outcomes of infections in the survivors of out-of-hospital cardiac arrest (OHCA) within the first 7 days after resuscitation. DESIGN AND SETTING: Retrospective cohort study in the intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: We enrolled 117 survivors of adult nontraumatic OHCA victims who survived more than 24 h between January 1999 and May 2004. We collected patients' demographics, the causes and initial electrocardiographic rhythm of cardiac arrest, and the process of cardiopulmonary resuscitation. The incidence, clinical presentations and outcomes of infections occurring in the first 7 days after resuscitation were evaluated. Variables were compared between the infected and noninfected patients. MEASUREMENTS AND RESULTS: Among our OHCA survivors asystole was the most common initial rhythm (66%). Eighty-three patients (71%) were found to have infection. Pneumonia was the most common infection (61%) followed by bacteremia (13%). Although the Gram-negative bacteria were responsible for most infections, the most commonly isolated organism was Staphylococcus aureus. The infection group had more patients with dementia and noncardiac causes of OHCA. The survival curves did not differ significantly between infection and non infection groups. CONCLUSIONS: Infections were common in OHCA survivors during the first 7 days. The most common responsible organisms were Gram-negative bacteria, and the most commonly isolated organism was S. aureus. Infections in the early stage after return of spontaneous circulation did not change the hospital mortality and hospitalization duration. PMID- 15803299 TI - Norepinephrine and metaraminol in septic shock: a comparison of the hemodynamic effects. AB - OBJECTIVE: To compare the effects of norepinephrine and metaraminol on hemodynamics in septic shock patients. DESIGN AND SETTING: Open-label, controlled clinical trial in the general intensive care unit of a university-affiliated hospital. PATIENTS AND PARTICIPANTS: Ten consecutive septic shock patients receiving norepinephrine to maintain the mean arterial pressure higher than 65 mmHg. INTERVENTIONS: Patients were monitored with pulmonary artery catheter and indirect calorimetry. At the baseline hemodynamic variables were obtained during norepinephrine infusion. Subsequently norepinephrine was replaced by metaraminol infusion in a dose sufficient to keep mean arterial pressure constant. After 20 min of stable arterial pressure a new set of measurement was repeated. MEASUREMENTS AND RESULTS: Mean arterial pressure did not differ significantly with norepinephrine or metaraminol; there was no relationship between the norepinephrine and metaraminol doses. Replacement norepinephrine with metaraminol did not modify hemodynamic variables; in particular there were no changes in heart rate, stroke volume index, pulmonary artery occlusion pressure, or oxygen consumption index. CONCLUSIONS: This study shows that metaraminol increases arterial pressure as does norepinephrine in septic shock patients. Despite similar effects of norepinephrine and metaraminol, there was no relationship between the dose of the two drugs. PMID- 15803300 TI - Bedside colonoscopy for critically ill patients with acute lower gastrointestinal bleeding. AB - OBJECTIVE: To determine the clinical impact of bedside colonoscopy for critically ill patients with acute lower gastrointestinal (GI) bleeding. DESIGN AND SETTING: A 3-year retrospective analysis (chart review). Medical intensive care unit (ICU) of a 1,312-bed tertiary-care center in Taiwan. PATIENTS AND PARTICIPANTS: Fifty five people undergoing bedside colonoscopy for lower GI bleeding that developed while in the ICU. INTERVENTIONS: Bedside colonoscopy. MEASUREMENTS AND RESULTS: Colonoscopy was successful in diagnosing the source of bleeding in 37 patients. Among them, colitis (15 patients, including ischemic, pseudomembranous, or radiation-induced) and acute hemorrhagic rectal ulcer (nine patients) were the most frequent confirmed causes. In seven patients, fresh blood was noticed above the colonoscopically accessible area and considered to originate from the small bowel. No adverse event was associated with colonoscopy. Spontaneous cessation of bleeding was noted in 29 (29/55, 53%) patients, whereas 16 (16/55, 29%) achieved endoscopic hemostasis. Ten (10/55, 18%) patients failed primary hemostasis or localization. Overall in-hospital mortality was 53% (29/55); however, hemorrhage related death occurred in only two patients. CONCLUSIONS: ICU patients with acute lower GI bleeding have distinctive causes. Bedside colonoscopy is effective for diagnosis in two-thirds of patients, but only a minority of them needs endoscopic hemostasis. PMID- 15803301 TI - Central venous-arterial carbon dioxide difference as an indicator of cardiac index. AB - OBJECTIVE: The mixed venous-arterial (v-a) pCO(2) difference has been shown to be inversely correlated with the cardiac index (CI). A central venous pCO(2), which is easier to obtain, may provide similar information. The purpose of this study was to examine the correlation between the central venous-arterial pCO(2) difference and CI. DESIGN: Prospective, cohort study. SETTING: Intensive care unit of an urban tertiary care hospital. PATIENTS AND PARTICIPANTS: Eighty-three consecutive intensive care unit patients. MEASUREMENTS: Simultaneous blood gases from the arterial, pulmonary artery (PA), and central venous (CV) catheters were obtained. At the same time point, cardiac indices were measured by the thermodilution technique (an average of three measurements). The cardiac indices obtained by the venous-arterial differences were compared with those determined by thermodilution. RESULTS: The correlation (R(2)) between the mixed venous arterial pCO(2) difference and cardiac index was 0.903 (p <0.0001), and the correlation between the central venous-arterial pCO(2) difference and cardiac index was 0.892 (p <0.0001). The regression equations for these relationships were natural log (CI)=1.837-0.159 (v-a) CO(2) for the PA and natural log (CI)=1.787-0.151 (v-a) CO(2) for the CV (p <0.0001 for both). The root-mean squared error for the PA and CV regression equations were 0.095 and 0.101, respectively. CONCLUSION: Venous-arterial pCO(2) differences obtained from both the PA and CV circulations inversely correlate with the cardiac index. Substitution of a central for a mixed venous-arterial pCO(2) difference provides an accurate alternative method for calculation of cardiac output. PMID- 15803302 TI - In vitro study and semiempirical model for aerosol delivery control during mechanical ventilation. AB - The object of this study was to evaluate in vitro the influence of various ventilatory parameters on the delivery of synchronized nebulization of terbutaline during mechanical ventilation and to determine a semiempirical model to control the quantity of aerosol delivered into the patient's lung. An ATOMISOR NL9 M jet nebulizer (La Diffusion Technique Francaise, France) was filled with terbutaline (Bricanyl, Astra-Zeneca, Sweden) and connected to the inspiratory line of a Horus ventilator (Taema, France). Nebulization was synchronized with the inspiratory phase. We assessed at the end of the endotracheal tube the quantity of terbutaline (terbutaline mass output) and the volume median diameter (VMD) by diffraction-laser method. There was a negative correlation between terbutaline mass output and inspiratory air flow ( r =-0.95, p <0.0001) and between VMD and inspiratory air flow ( r =-0.96, p <0.0001). Moreover, positive end-expiratory pressure levels between 0 cm and 8 cm of water did not significantly change the terbutaline output mass ( p =0.22). Total nebulization time and terbutaline mass output calculated by the mathematical model showed good agreement with experimental data. In conclusion, our semiempirical model allows calculation of the duration of the nebulization required to deliver a given mass of terbutaline into patient lungs. PMID- 15803305 TI - Study of the suitability of HNO3 and HCl as extracting agents of mercury species in soils from cinnabar mines. AB - The aim of this study was to compare the influence and feasibility of two common extracting agents (50% v/v HCl and 50% v/v HNO3) on the leaching of Hg from soils. The solubility of a number of Hg species in each acid solution was evaluated under selected conditions. Most species were quantitatively dissolved in both acids with the exception of HgS. The application of both acid solutions to a soil sample from the Almaden mining area provided different recoveries of Hg: about 5% in 50% v/v HNO3 and 50% in 50% v/v HCl. The following experiments were designed and developed in order to evaluate the matrix influence on HgS solubility and leaching: (1) study of the solubility of HgS in the presence of different potential interfering compounds such as FeCl3, KCl, KI, Fe2O3, CuSO4, FeSO4, MnO2 and NaNO3; (2) study of the recovery of HgS spiked in soil samples; (3) study of the extraction process in soil samples spiked with the critical interfering compounds. Results showed the existence of a greater matrix influence with the HCl solution, since much higher Hg recoveries were obtained with this reagent. In addition, the presence of nitrates and Mn oxides drastically promotes the solubility of HgS in an HCl solution. On the other hand, halide compounds drastically enhanced the extractability of Hg in the HNO3 and they must be considered as potential interfering compounds when this acid solution is used as extracting agent. In summary, neither acid is totally free of matrix effects from common soil constituents; conclusions about mercury mobility resulting from the general application of these extraction procedures must therefore be made with caution. PMID- 15803307 TI - An electro-osmotic micro-pump based on monolithic silica for micro-flow analyses and electro-sprays. AB - A high-pressure electro-osmotic micro-pump fabricated by a sol-gel process is reported as a fluid-driving unit in a flow-injection analysis (FIA) system. The micro FIA system consists of a monolithic micro-pump on a glass slide (2.5 x 7.5 cm), a micro-injector, and a micro-sensor (2.5 x 1.5 cm). The monolithic silica matrix has a continuous skeleton morphology with micrometer-sized through-pores. The micrometer-size pores with a large negative surface charge density build up a large pressure under a DC electric field to drive fluid through the downstream units. A novel Nafion joint for the downstream cathode eliminates flow into the electrode reservoir and further enhances pressure build-up. The measured pump pressure curve indicated a maximum pressure of 0.4 MPa at flow rate of 0.4 microL min(-1) at 6 kV. Despite the large voltage, the small current transmission area through the monolith produced a negligible current (less than 100 microA) that did not generate bubbles or ion contaminants. The flow rate can be precisely controlled in the range 200 nL to 2.5 microL min(-1) by varying the voltage from 1 to 6 kV. The high pump pressure and the large current-free DC field also enabled the pump to act as an electro-spray interface with a downstream analytical instrument. PMID- 15803303 TI - Quality of life in adult survivors of critical illness: a systematic review of the literature. AB - OBJECTIVE: To determine how the quality of life (QOL) of intensive care unit (ICU) survivors compares with the general population, changes over time, and is predicted by baseline characteristics. DESIGN: Systematic literature review including MEDLINE, EMBASE, CINAHL and Cochrane Library. Eligible studies measured QOL > or = 30 days after ICU discharge using the Medical Outcomes Study 36-item Short Form (SF-36), EuroQol-5D, Sickness Impact Profile, or Nottingham Health Profile in representative populations of adult ICU survivors. Disease-specific studies were excluded. MEASUREMENTS AND RESULTS: Of 8,894 citations identified, 21 independent studies with 7,320 patients were reviewed. Three of three studies found that ICU survivors had significantly lower QOL prior to admission than did a matched general population. During post-discharge follow-up, ICU survivors had significantly lower QOL scores than the general population in each SF-36 domain (except bodily pain) in at least four of seven studies. Over 1-12 months of follow-up, at least two of four studies found clinically meaningful improvement in each SF-36 domain except mental health and general health perceptions. A majority of studies found that age and severity of illness predicted physical functioning. CONCLUSIONS: Compared with the general population, ICU survivors report lower QOL prior to ICU admission. After hospital discharge, QOL in ICU survivors improves but remains lower than general population levels. Age and severity of illness are predictors of physical functioning. This systematic review provides a general understanding of QOL following critical illness and can serve as a standard of comparison for QOL studies in specific ICU subpopulations. PMID- 15803309 TI - Quantitative determination of capsaicinoids by liquid chromatography-electrospray mass spectrometry. AB - Eight naturally occurring capsaicinoids have been determined in Capsicum by use of high-purity standards, with norcapsaicin as an internal standard. The solid standards were rigorously checked for purity. The sensitivity of electrospray ionization (ESI), atmospheric-pressure chemical ionization (APCI), and coordination ion-spray (CIS; with silver) toward the capsaicinoids were measured and compared. The highest sensitivity was found for positive-ion ESI. Method validation of the liquid chromatography-ESI-mass spectrometry (LC-ESI-MS) determination is reported, including tests for repeatability (4%), detection limit (5 pg injected), linear range (20-6 ng injected), quantitation (excellent linearity; < 2% relative standard deviation), and recovery (99-103%). The major and minor capsaicinoids in a commercial plant extract and in chili pepper fruits were quantified. PMID- 15803310 TI - Reversed-phase liquid chromatography and argentation chromatography of the minor capsaicinoids. AB - An investigation of the liquid chromatography of the minor capsaicinoids in a commercial capsaicinoid mixture is reported. Twelve stationary phases including C8, C18, C30, phenyl, and cation-exchange chemistries were examined in combination with isocratic aqueous methanol and aqueous acetonitrile mobile phases. A phenyl stationary phase and aqueous acetonitrile mobile phase baseline resolved 7 of 11 capsaicinoids, and selected ion chromatograms (LC-ESI-MS) demonstrated this was the most effective reversed-phase separation. Argentation chromatography with an alkyl or phenyl column and aqueous silver nitrate-methanol mobile phase revealed the presence of the 6-ene-8-methyl and 6-ene-9-methyl homocapsaicin isomers and the absence of 7-ene-9-methyl homocapsaicin. A mixed phenyl-cation-exchange stationary phase (charged with silver ion) enabled unique and useful separations of the capsaicinoids. PMID- 15803311 TI - Biofiltration of volatile organic compounds. AB - The removal of volatile organic compounds (VOCs) from contaminated airstreams has become a major air pollution concern. Improvement of the biofiltration process commonly used for the removal of odorous compounds has led to a better control of key parameters, enabling the application of biofiltration to be extended also to the removal of VOCs. Moreover, biofiltration, which is based on the ability of micro-organisms to degrade a large variety of compounds, proves to be economical and environmentally viable. In a biofilter, the waste gas is forced to rise through a layer of packed porous material. Thus, pollutants contained in the gaseous effluent are oxidised or converted into biomass by the action of microorganisms previously fixed on the packing material. The biofiltration process is then based on two principal phenomena: (1) transfer of contaminants from the air to the water phase or support medium, (2) bioconversion of pollutants to biomass, metabolic end-products, or carbon dioxide and water. The diversity of biofiltration mechanisms and their interaction with the microflora mean that the biofilter is defined as a complex and structured ecosystem. As a result, in addition to operating conditions, research into the microbial ecology of biofilters is required in order better to optimise the management of such biological treatment systems. PMID- 15803312 TI - Characterization of Rat8 localization and mRNA export in Saccharomyces cerevisiae during the brewing of Japanese sake. AB - Ethanol affects the nuclear export of mRNA in a similar way to heat shock in Saccharomyces cerevisiae. We recently reported that the nuclear accumulation of Rat8 caused by ethanol stress correlates well with blocking of the export of bulk poly(A)(+) mRNA. Here, we characterize the localization of Rat8 and bulk poly(A)(+) mRNA in sake (Japanese rice wine) yeast during the brewing of sake. In wine must and synthetic dextrose medium, sake yeast showed the same responses to ethanol regarding changes in the localization of Rat8 as wine yeast and a laboratory strain: i.e., cells began the nuclear accumulation of Rat8 at an ethanol concentration of 6% and completed it at 9%. In contrast, during the sake brewing process, sake yeast showed unique phenomena: i.e., cells did not start the nuclear accumulation of Rat8 until the ethanol concentration of the sake mash reached around 12% and they showed a normal localization of Rat8 around the nuclear envelope at the late stage of fermentation. These results provide new information about the transport of mRNA in yeast cells during actual alcoholic fermentation. PMID- 15803313 TI - Fast determination of operational stability of the soluble acetylacetone-cleaving enzyme Dke1 in an enzyme membrane reactor. AB - The main aim of this study was the determination of the operational stability of soluble Dke1 (EC 1.13.11.50) in an enzyme membrane reactor. In order to calculate the half-life of soluble Dke1, the K (M) of oxygen must be known. The determination of this constant was done using progress curve analysis (K (M) = 260 micromol l(-1)). In a next step, the reactor system was studied by building a mathematical model for calculation of the reactor system, using Berkeley Madonna ver. 8.0.1 software. After that, the determination of the half-life of Dke1 under operational conditions at different temperatures (5, 10, 15, 25, 30, 35 degrees C) was performed. The quantitative criterion for stability was the value of the first-order rate constant of monomolecular inactivation. The experiments showed that soluble Dke1 is poorly stable. The half-life ranged from 308 min at 5 degrees C to 9 min at 35 degrees C. This method for determining the half-life is quite applicable for enzymes which are poorly stable. In addition, both the storage stability and the operational stability can be determined. PMID- 15803314 TI - Effects of perindopril on cardiac sympathetic nerve activity in patients with congestive heart failure: comparison with enalapril. AB - PURPOSE: The production of aldosterone in the heart is suppressed by the angiotensin-converting enzyme (ACE) inhibitor perindopril in patients with congestive heart failure (CHF). Moreover, perindopril has been reported to have more cardioprotective effects than enalapril. MATERIALS AND METHODS: Forty patients with CHF [left ventricular ejection fraction (LVEF) <45%; mean 33+/-7%] were randomly assigned to perindopril (2 mg/day; n = 20) or enalapril (5 mg/day; n = 20). All patients were also treated with diuretics. The delayed heart/mediastinum count (H/M) ratio, delayed total defect score (TDS) and washout rate (WR) were determined from 123I-meta-iodobenzylguanidine (MIBG) images, and plasma brain natriuretic peptide (BNP) concentrations were measured before and 6 months after treatment. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV) and LVEF were also determined by echocardiography. RESULTS: After treatment, in patients receiving perindopril, TDS decreased from 39+/-10 to 34+/-9 (P < 0.01), H/M ratios increased from 1.62+/ 0.27 to 1.76+/-0.29 (P < 0.01), WR decreased from 50+/-14% to 42+/-14% (P < 0.05) and plasma BNP concentrations decreased from 226+/-155 to 141+/-90 pg/ml (P < 0.0005). In addition, the LVEDV decreased from 180+/-30 to 161+/-30 ml (P < 0.05) and the LVESV decreased from 122+/-35 to 105+/-36 ml (P < 0.05). Although the LVEF tended to increase, the change was not statistically significant (from 33+/ 8% to 36+/-12%; P = NS). On the other hand, there were no significant changes in these parameters in patients receiving enalapril. CONCLUSION: Plasma BNP concentrations, 123I-MIBG scintigraphic and echocardiographic parameters improved after 6 months of perindopril treatment. These findings indicate that perindopril treatment can ameliorate the cardiac sympathetic nerve activity and the left ventricular performance in patients with CHF. PMID- 15803315 TI - Bone marrow features of diagnostic impact in erythrocytosis. AB - Controversy continues to persist about the role of histopathology regarding diagnosis of polycythemia vera (PV). For this reason, a clinicopathological study was performed on 334 patients presenting with a sustained borderline to marked erythrocytosis (hemoglobin >17 g/dl in men and >15 g/dl in women). The aim was to elucidate the discriminating impact of bone marrow biopsy examinations in an independent fashion from laboratory parameters. According to morphological findings based on a semiquantitative evaluation of standardized features, cellularity, megakaryocytes (quantity, size, pleomorphous aspect, clustering, nuclear lobulation), eosinophils, cellular debris, perivascular plasmacytosis and iron-laden macrophages exerted a distinctive value. Comparison with clinical data and follow-up revealed that in only 13 patients (4%), histopathology failed to differentiate clearly between PV (208 patients) and secondary polycythemias (113 patients). In conclusion, certain sets of morphological parameters allow a distinction between autonomous and reactive polycythemias and therefore enhance significantly diagnostic validity. PMID- 15803316 TI - A patient with anophthalmia, hemifacial microsomia, and hemoglobin anti-Lepore. PMID- 15803317 TI - Egg-laying-hormone immunoreactivity in the neural ganglia and ovary of Haliotis asinina Linnaeus. AB - Immunoreactivity against the abalone egg-laying hormone (aELH) was detected in the fine granules of type 1 and 2 neurosecretory (NS) cells, neurites in the neuropil, and blood sinuses in the connective tissue sheath of the cerebral, pleuropedal, and visceral ganglia of the tropical abalone, Haliotis asinina Linnaeus. The number of positive NS cells, and the intensity of staining in the ganglia, varied and might be related to the stage of ovarian cycle. At any stage, positive cells were most numerous in the pleuropedal, and least numerous in the visceral ganglion. In addition, several cells of the statocyst and associated nerves also exhibited the immunoreactivity. In the ovary, the most intense reactivity was detected in the follicular and granular cells adjacent to mature oocytes, in the trabeculae and the ovarian capsule. The cytoplasm of mature oocytes was also moderately stained. The results indicate that the cerebral, pleuropedal, and visceral ganglia are the main sites of aELH-producing cells. The ovary may also produce aELH locally. PMID- 15803318 TI - Prevalence of concomitant sleep disorders in patients with obstructive sleep apnea. AB - We determined the prevalence of concomitant sleep disorders in patients with a primary diagnosis of obstructive sleep apnea (OSA). We retrospectively analyzed 643 patients, aged > or =18, with a primary diagnosis of OSA, evaluated by sleep specialists, in whom clinical and polysomnographic data were derived using standardized techniques by reviewing data from a standardized database and clinical charts. Concomitant sleep disorders were listed according to the International Classification of Sleep Disorders (American Academy of Sleep Medicine, 2000). The mean age was 48.5+/-13.5 years and 55% were male. Racial distributions were African-Americans 51.8% and Caucasian 47%. Indices of disordered breathing were respiratory disturbance index 32.4+/-30.4/h sleep and time <90% O(2) saturation 44.5+/-81.6 min. Thirty-one percent of patients had a concomitant sleep disorder. The most common were inadequate sleep hygiene (14.5%) and periodic limb movement disorder (PLMD, 8.1%). Of patients with other sleep disorders, 66.8% had treatment initiated for these disorders. Predictors of inadequate sleep hygiene (logistic regression) were: age (each decade OR=0.678, P=0.000000), gender (for M, OR=0.536), and the presence of at least one other major system disorder (OR=2.123, P=0.0015). Predictors of PLMD were: age (each decade OR=0.794, P=0.0005), gender (for M, OR=0.433, P=0.004), and total sleep time (for each 10 min, OR=0.972, P=0.0013). We conclude that approximately one third of patients with sleep apnea have another identifiable sleep disorder, usually requiring treatment. This suggests that practitioners evaluating and treating sleep apnea ought to be prepared to deal with other sleep disorders as well. PMID- 15803319 TI - Identification of an active transposon in intact rice plants. AB - A transposable element that is active in intact plants has been identified in rice (Oryza sativa L.). The 607-bp element itself, termed nonautonomous DNA-based active rice transposon (nDart), has no coding capacity. It was found inserted in the gene encoding Mg-protoporphyrin IX methyltransferase in a chlorophyll deficient albino mutant isolated from backcross progeny derived from a cross between wild-type japonica varieties. The nDart has 19-bp terminal inverted repeats (TIRs) and, when mobilized, generates an 8-bp target-site duplication (TSD). At least 13 nDart elements were identified in the genome sequence of the japonica cultivar Nipponbare. Database searches identified larger elements, termed DNA-based active rice transposon (Dart) that contained one ORF for a protein that contains a region with high similarity to the hAT dimerization motif. Dart shares several features with nDart, including identical TIRs, similar subterminal sequences and the generation of an 8-bp TSD. These shared features indicate that the nonautonomous element nDart is an internal deletion derivative of the autonomous element Dart. We conclude that these active transposon systems belong to the hAT superfamily of class II transposons. Because the transposons are active in intact rice plants, they should be useful tools for tagging genes in studies of functional genomics. PMID- 15803320 TI - Regulation of homologous integration in yeast by the DNA repair proteins Ku70 and RecQ. AB - The product of the BLM gene, which is mutated in Bloom syndrome in humans, and the Saccharomyces cerevisiae protein Sgs1 are both homologous to the Escherichia coli DNA helicase RecQ, and have been shown to be involved in the regulation of homologous recombination. Mutations in these genes result in genome instability because they increase the incidence of deletions and translocations. We present evidence for a genetic interaction between SGS1 and YKU70, which encodes the S. cerevisiae homologue of the human DNA helicase Ku70. In a yku70 mutant background, sgs1 mutations increased sensitivity to DNA breakage induced either by treatment with camptothecin or by the expression of the restriction enzyme EcoRI. The yku70 mutation caused a fourfold increase in the rate of double-strand break (DSB)-induced target integration as that seen in the sgs1 mutant. The combination of yku70 and sgs1 mutations additively increased the rate of the targeted integration, and this effect was completely suppressed by deletion of RAD51. Interestingly, an extra copy of YKU70 partially suppressed the increase in targeted integration seen in the sgs1 single mutant. These results suggest that Yku70 modulates the repair of DSBs associated with homologous recombination in a different way from Sgs1, and that the inactivation of RecQ and Ku70 homologues may enhance the frequency of gene targeting in higher eukaryotes. PMID- 15803324 TI - [Benefits and risks of nitrate preparations in the treatment of angina pectoris]. PMID- 15803325 TI - [Clinical study of infectious diseases. An interdisciplinary challenge in medicine]. PMID- 15803323 TI - Modulation of higher-plant NAD(H)-dependent glutamate dehydrogenase activity in transgenic tobacco via alteration of beta subunit levels. AB - Glutamate dehydrogenase (GDH; EC 1.4.1.2-1.4.1.4) catalyses in vitro the reversible amination of 2-oxoglutarate to glutamate. In vascular plants the in vivo direction(s) of the GDH reaction and hence the physiological role(s) of this enzyme remain obscure. A phylogenetic analysis identified two clearly separated groups of higher-plant GDH genes encoding either the alpha- or beta-subunit of the GDH holoenzyme. To help clarify the physiological role(s) of GDH, tobacco (Nicotiana tabacum L.) was transformed with either an antisense or sense copy of a beta-subunit gene, and transgenic plants recovered with between 0.5- and 34 times normal leaf GDH activity. This large modulation of GDH activity (shown to be via alteration of beta-subunit levels) had little effect on leaf ammonium or the leaf free amino acid pool, except that a large increase in GDH activity was associated with a significant decrease in leaf Asp (~51%, P=0.0045). Similarly, plant growth and development were not affected, suggesting that a large modulation of GDH beta-subunit titre does not affect plant viability under the ideal growing conditions employed. Reduction of GDH activity and protein levels in an antisense line was associated with a large increase in transcripts of a beta-subunit gene, suggesting that the reduction in beta-subunit levels might have been due to translational inhibition. In another experiment designed to detect post-translational up-regulation of GDH activity, GDH over-expressing plants were subjected to prolonged dark-stress. GDH activity increased, but this was found to be due more likely to resistance of the GDH protein to stress induced proteolysis, rather than to post-translational up-regulation. PMID- 15803326 TI - Quantitative echographic analysis of photochemotherapy on systemic sclerosis skin. AB - Systemic sclerosis (SSc) is a connective tissue disease of unknown etiology, in which development of fibrosis, vascular insufficiency and inflammatory processes are prominent in the skin as well as in other organs. We studied the effect of photochemotherapy with quantitative echography in SSc. Dermal echo intensity and dermal thickness was measured using high-frequency dermal echography before and after therapy. The dermal echo intensity after photochemotherapy (33.51+/-9.34) significantly increased than that before therapy (21.23+/-6.00) (P < 0.01), while dermal thickness (1.20+/-0.20) significantly decreased than that before therapy (1.38+/-0.18) (P < 0.05). Photochemotherapy was more likely to improve dermal edema, not fibrosis, because echo intensity after treatment was significantly elevated with that before treatment in patients with edema. Quantitative echographic analysis was concluded to be a reliable method in evaluating the change of skin edema in SSc. PMID- 15803327 TI - Ceramide profiles of the uninvolved skin in atopic dermatitis and psoriasis are comparable to those of healthy skin. AB - Ceramides are sphingolipids consisting of sphingoidbases, which are amide-linked to fatty acids. In the stratum corneum, they represent the major constituent of the free extractable intercellular lipids and play a significant role in maintaining and structuring the water permeability barrier of the skin. Using thin layer chromatography, which represents the method of the first choice in analyzing the stratum corneum ceramides, at least seven classes can be distinguished. Each ceramide class contains various species, which have the same head group and different chain lengths. As in many other skin disorders, atopic dermatitis and psoriasis show derangements in content and profile of the ceramides. Such derangements were reported for both the lesional involved as well as for the normal-appearing uninvolved skin. In this study, we focused on investigating the stratum corneum ceramides of the uninvolved skin in atopic dermatitis and psoriasis patients compared to healthy skin. The aim of the investigations was to explore possible significant and specific differences which can be accomplished for purposes of early diagnostics. The skin lipids were collected by means of an in vivo topical extraction procedure using an extraction mixture consisting of n-hexane and ethanol, (2:1). An automated multiple development-high performance thin layer chromatography (AMD-HPTLC) method with photodensitometric detection were applied to separate the ceramides and to estimate their contents. For studying their molecular profile within each ceramide class, a new method of normal phase HPLC with atmospheric pressure chemical ionization mass spectrometry were used. The results obtained by AMD HPTLC exposed no significant alterations regarding the relative composition of the major stratum corneum lipids and primarily the ceramides. In addition, the mass spectrometric profiles within each ceramide class were similar in the patients and the healthy control subjects. In conclusion, this study revealed that the normal-appearing uninvolved skin of atopic dermatitis and psoriasis patients does not prove significant or specific deficiencies with respect to the free extractable major stratum corneum lipids and mainly the ceramides, when compared to healthy skin. Thus, they cannot be used for diagnostic purposes. Furthermore, our data are not consistent with the concept that impairments in the ceramide composition represent an obligate etiologic factor for both diseases. PMID- 15803328 TI - Increased expression of p53 and p21 (Waf1/Cip1) in the lesional skin of bleomycin induced scleroderma. AB - Systemic sclerosis (SSc) is a connective tissue disorder characterized by excessive deposition of extracellular matrix in the affected skin as well as various internal organs, vascular injury and immune abnormality; however, the etiology of SSc remains still unknown. We previously established an experimental mouse model for scleroderma by repeated local injections of bleomycin, a DNA damaging agent. In this study, we examined the induction of apoptosis and the expression of p53, p21 (Waf1/Cip1), and proliferating cell nuclear antigen (PCNA) in the lesional skin following bleomycin exposure in this model. Dermal sclerosis was induced by alternate day's injections of bleomycin for 4 weeks. TUNEL assay showed that apoptotic cells began to appear at 1 week after bleomycin exposure, and were prominently detected at 3-4 weeks. Immunohistochemical examination showed increased expression of p53 and p21 mainly in the infiltrating mononuclear cells at 2 weeks after bleomycin treatment. Bleomycin treatment markedly enhanced PCNA expression at 1-2 weeks, mainly in mesenchyme, as compared with control phosphate buffered saline treatment. Reverse transcriptase-polymerase chain reaction analysis showed that the expression of p53 and p21 mRNA was concurrently upregulated at 1-2 weeks after bleomycin treatment. Taken together, coordinate increased levels of p53 and p21 preceded the maximal induction of apoptosis and dermal sclerosis. Our findings suggest that apoptotic processes are involved in the pathophysiology of bleomycin-induced scleroderma, which may be mediated, in part, by the upregulation of p53 and p21. PMID- 15803329 TI - Anti-cooperativity in diffusion-controlled reactions with pairs of anisotropic domains: a model for the antigen-antibody encounter. AB - The encounter between anisotropic agents in diffusion-controlled reactions is a topic of very general relevance in chemistry and biology. Here we introduce a simplified model of encounter of an isotropic molecule with a pair of partially reacting agents and apply it to the encounter reaction between an antibody and its antigen. We reduce the problem to the solution of dual series relations, which can be solved iteratively, yielding the exact solution for the encounter rate constant at any desired order of accuracy. We quantify the encounter effectiveness by means of a simple indicator and show that the two binding centers systematically behave in an anti-cooperative fashion. However, we demonstrate that a reduction of the binding active sites allows the composite molecule to recover binding effectiveness, in spite of the overall reduction of the rate constant. In addition, we provide a simple formula that enables one to calculate the anti-cooperativity as a function of the size of the binding site for any values of the separation between the two active lobes and of the antigen size. Finally, some biological implications of our results are discussed. PMID- 15803330 TI - Validation of the 53A6 GROMOS force field. AB - The quality of biomolecular dynamics simulations relies critically on the force field that is used to describe the interactions between particles in the system. Force fields, which are generally parameterized using experimental data on small molecules, can only prove themselves in realistic simulations of relevant biomolecular systems. In this work, we begin the validation of the new 53A6 GROMOS parameter set by examining three test cases. Simulations of the well studied 129 residue protein hen egg-white lysozyme, of the DNA dodecamer d(CGCGAATTCGCG)(2), and a proteinogenic beta(3)-dodecapeptide were performed and analysed. It was found that the new parameter set performs as well as the previous parameter sets in terms of protein (45A3) and DNA (45A4) stability and that it is better at describing the folding-unfolding balance of the peptide. The latter is a property that is directly associated with the free enthalpy of hydration, to which the 53A6 parameter set was parameterized. PMID- 15803332 TI - [Cystic liver lesions with unspecified upper abdominal pain]. PMID- 15803331 TI - Insulin sensitivity, proinsulin and insulin as predictors of coronary heart disease. A population-based 10-year, follow-up study in 70-year old men using the euglycaemic insulin clamp. AB - AIMS/HYPOTHESIS: The association between CHD and insulin sensitivity (Si) measured by the euglycaemic insulin clamp has not been examined previously. Earlier studies found a relationship between CHD and elevated plasma insulin, an analysis that may have been confounded by co-determination of proinsulin, which has evolved as a stronger predictor of CHD. The aim was to determine the longitudinal relationships between Si, intact proinsulin, 32-33 split proinsulin, specific insulin and subsequent CHD. METHODS: This was a population-based cohort study of 815 men in Uppsala, Sweden, aged 70 years at baseline with a follow-up of up to 10 years. Baseline insulin sensitivity was determined by euglycaemic insulin clamp. Fasting proinsulin, 32-33 split proinsulin and specific insulin concentrations were analysed using specific two-site immunometric assays. CHD was taken as diagnosed, if stated (in the event of death) on the Cause of Death Registry, or for subjects hospitalised for the first time with CHD, if CHD was recorded in the Hospital-Discharge Registry. The associations were analysed using Cox's proportional hazards, presented as hazard ratios (HRs) with their 95% CIs for a one-SD increase in the predictor. RESULTS: In multivariate analysis, Si (HR:0.80, CI:0.65-0.97) adjusted for serum cholesterol, systolic blood pressure, fasting plasma glucose, BMI and smoking predicted CHD. Intact proinsulin (HR:1.18, CI:1.01-1.38), adjusted as the model above, predicted CHD, whereas 32 33 split proinsulin (HR:1.13, CI:0.95-1.35) or specific insulin (HR:1.07, CI:0.89 1.30) did not. CONCLUSIONS/INTERPRETATION: Insulin resistance measured by the euglycaemic insulin clamp predicts subsequent CHD in elderly men. Proinsulin provides a better prediction of CHD than insulin. PMID- 15803333 TI - [Health status and health-related quality of life. A facet in the evaluation of outcomes in orthopaedic surgery]. AB - Traditional methods for evaluating the outcome of treatment in orthopaedic surgery include the assessment of clinical status, pain and radiography. For more than two decades, there has been an increasing interest in the use of health status measures in clinical medicine. Outcome evaluation should not only consider clinical and radiographic parameters but also the patients' personal views on their health status and quality of life. Questionnaires developed from basic principles allow the measurement of these views objectively, as well as the comparison of data between groups. The current paper primarily addresses the orthopaedic surgeon and provides a review of quality of life and health-status measures commonly used in orthopaedic outcome studies. Generic and disease specific measures of health status can be used in orthopaedic surgery to complement an outcome assessment. Thereby, the personal view of the patient becomes an objective part of the evaluation. PMID- 15803335 TI - Enlarged wandering spleen treated with hemisplenectomy and fixation of the residual spleen. AB - A case of enlarged wandering spleen presenting as an abdominal mass in a 7-year old girl is reported. The diagnosis was performed by ultrasonography and confirmed by aortography. Treatment consisted of an elective partial splenectomy and fixation of the residual spleen to the left hemidiaphragm. The patient's symptoms regressed, and now, after more than 2 years, she is doing well. Two years postoperatively scintigraphy demonstrated a good captation of radionuclide by a normal-sized spleen at the left hypochondrium. The results obtained with the procedure that was adopted for treating this case of wandering spleen allow the authors to state that hemispleen fixation is a valid option for treating symptomatic enlarged wandering spleen. PMID- 15803334 TI - Structural differences between the ready and unready oxidized states of [NiFe] hydrogenases. AB - [NiFe] hydrogenases catalyze the reversible heterolytic cleavage of molecular hydrogen. Several oxidized, inactive states of these enzymes are known that are distinguishable by their very different activation properties. So far, the structural basis for this difference has not been understood because of lack of relevant crystallographic data. Here, we present the crystal structure of the ready Ni-B state of Desulfovibrio fructosovorans [NiFe] hydrogenase and show it to have a putative mu-hydroxo Ni-Fe bridging ligand at the active site. On the other hand, a new, improved refinement procedure of the X-ray diffraction data obtained for putative unready Ni-A/Ni-SU states resulted in a more elongated electron density for the bridging ligand, suggesting that it is a diatomic species. The slow activation of the Ni-A state, compared with the rapid activation of the Ni-B state, is therefore proposed to result from the different chemical nature of the ligands in the two oxidized species. Our results along with very recent electrochemical studies suggest that the diatomic ligand could be hydro-peroxide. PMID- 15803336 TI - Secondary closure of a giant omphalocele by translation of the muscular layers: a new method. AB - The current report describes a case of an infant girl with a giant omphalocele in whom a new surgical technique was used for closing the abdominal wall after epithelialization of the omphalocele for 16 months. The technique used was translation of the muscular layers of the abdominal wall. The functional and cosmetic results appear superior compared with other suggested treatments used for this abdominal wall defect. PMID- 15803337 TI - High-resolution magic-angle spinning 13C NMR spectroscopy of cerebral tissue. AB - Monitoring the metabolism of (13)C-labelled substrates by biological tissues allows both the rate of metabolism and the relative importance of metabolic pathways to be determined. In this study high-resolution magic-angle spinning (HRMAS) (13)C NMR spectroscopy is assessed as a technique for determining the labelling of metabolites in brain slices. Freshly prepared rat brain slices were superfused in isotonic salt solution containing [1-(13)C] glucose. HRMAS (1)H and (13)C NMR spectra were acquired of the slices ( approximately 10 mg) at 3 degrees C. Using (1)H NMR spectroscopy it was demonstrated that the concentration of key metabolites indicative of metabolic degradation, including N-acetyl aspartate and lactate, did not change significantly across the approximately 11 h time period required for (13)C NMR spectra. The approach produced high-resolution spectra of intact tissue with the labelling patterns of tissues being indicative of both labelling via pyruvate dehydrogenase found in both neuronal and glial cells, and pyruvate carboxylase, found only within glial cells. This approach is a versatile tool for monitoring the compartmentation of metabolites directly, and will also allow the investigation of aqueous and lipid metabolites simultaneously. PMID- 15803338 TI - The Society for Pediatric Radiology 48th Annual Meeting and Postgraduate Course, New Orleans, Louisiana, May 3-7, 2005. Abstracts. PMID- 15803346 TI - Power-assisted lipoplasty versus traditional suction-assisted lipoplasty: comparative evaluation and analysis of output. PMID- 15803347 TI - Altering first impressions following facial plastic surgery. PMID- 15803348 TI - Mammary compliance: an objective measurement of capsular contracture. PMID- 15803349 TI - Complications associated with isotretinoin use after rhinoplasty. AB - BACKGROUND: This study aimed to evaluate the association between the postoperative use of isotretinoin and the formation of nasal tip deformities after rhinoplasty. METHODS: A retrospective study investigated a case series of patients who presented with nasal tip deformities subsequent to the use of isotretinoin after rhinoplasty. Patients who had taken isotretinoin after rhinoplasty were identified from a single surgeon's case log. Clinic charts and operative reports were reviewed. Predisposing factors and time intervals to complications were identified. RESULTS: Three patients were found to have undergone isotretinoin therapy after rhinoplasty. Nasal tip deformities included bossa formation, asymmetry, and prominence of a composite graft. All three patients required subsequent procedures to repair soft tissue nasal tip deformities. In all three cases, isotretinoin was started within 2 years of the primary rhinoplasty. The nasal tip deformities were first observed within 6 months after isotretinoin was started. CONCLUSIONS: Three cases are described in which postoperative use of isotretinoin was associated with nasal tip deformities after rhinoplasty. Further studies are needed to investigate this possible causative relationship and to describe the risks involved. PMID- 15803350 TI - Repetitive reconstructive rhinoplasty. AB - Plastic surgeons have the appropriate knowledge and clinical experience to perform a rhinoplasty. In the past 8 years, the authors have received for consultation some patients who had already undergone more than one rhinoplasty. As this shows, the specialist must be familiarized with the surgical technique specific for each case and its respective terminology. The authors propose the use of "repetitive reconstructive rhinoplasty" in referring to all the patients who for different reasons come to the specialist for a new rhinoplasty after a minimum of three surgeries performed previously. It must be emphasized that the plastic surgeon assumes the responsibility for choosing to perform repetitive reconstructive rhinoplasty, incurring the legal risk that the aesthetic or functional results may not satisfy the patient's expectations. One of the principal reasons for introducing repetitive reconstructive rhinoplasty emphasizes the "reconstructive" rather than the "aesthetic" role, whereas the legislation in many countries interprets the word "aesthetic" to imply an obligation for results that approximate perfection. PMID- 15803351 TI - Carbon dioxide laser resurfacing with fast recovery. AB - ABTRACT: Long sun exposure, in addition to ozone layer damage, produces structural damase to the normal skin. Injury to the dermal collagen and elastic fiber results in facial wrinkles. Photodamage to the skin is one of the most common sources of concern for patients visiting the plastic surgeon or dermatologist. Over the years, many alternative solutions have been developed. CO2 laser treatment is one of the alternatives bringing unique benefits and satisfactory results for both patient and surgeons. However, the initial problems of emotional discomfort, prolonged postoperative recovery and delayed return to normal activities have made patients reluctant to accept this method. This article discusses single-pass CO2 laser resurfacing with lower energy. Also, it proposes a technique that does not use wet gauze to remove the surface of the skin. This technique is applied in combination with an intensive skin care treatment. Different authors propose a single pass of CO2 laser with excellent results. With the reported method, identical long-lasting benefits are achieved, but the post-operative time is shorter. PMID- 15803352 TI - Can aesthetic facial reconstruction be judged in black and white? PMID- 15803353 TI - Evaluation of results from reduction mammaplasty: relief of symptoms and patient satisfaction. AB - Reduction mammoplasty is one of the most common operations performed in plastic surgery clinics. Although patients present with functional symptoms, the aim of treatment usually is obtaining aesthetic satisfaction. This retrospective study evaluated charts and operation notes of patients and a self-assessment questionnaire. A detailed questionnaire form was sent to 121 patients who had undergone reduction mammaplasty at the Dokuz Eylul University Medical Faculty Plastic, Reconstructive, and Aesthetic Surgery Clinic between 1991 and 2001. The questionnaire asked the subjects about pre- and postoperative, physical, and psychosocial symptoms, as well as operative satisfaction. The results were evaluated with respect to different techniques. The findings showed that reduction mammaplasty remains a very satisfying procedure for most of the patients undergoing this operation, as indicated by the high rate of patient satisfaction. The limited number of studies in this area and the lack of groups conducting such work in our society indicates the importance of similar study. PMID- 15803354 TI - Restoration of the gluteal fold by a deepithelialized skin flap: preliminary observations. PMID- 15803355 TI - Eyelid aging: the historical evolution of its management. AB - The evolution of blepharoplasties is reviewed. By better understanding the mechanism of herniated lower eye lid fat pads, one can understand the interrelated enophthalmia, tear trough deformities, and sunken upper lid. The authors use a technique of relocating the herniated fat pad that reverses this process. All of this is described in detail. PMID- 15803356 TI - A meta-analysis of FDG-PET for the evaluation of breast cancer recurrence and metastases. AB - OBJECTIVE: To evaluate the diagnostic performance of 18F-2-deoxy-2-fluoro-D glucose-positron emission tomography (FDG-PET) in the evaluation of breast cancer recurrence and metastases. DATA SOURCES: Articles published in medical journals during January 1995-June 2004 were identified by a systematic Medline search, supplemented by a manual search of the references listed in original and review articles. STUDY SELECTION: Studies that evaluated FDG-PET with a dedicated camera for the diagnosis of breast cancer recurrence or metastases, and reporting sufficient data to permit calculation of sensitivity and specificity, were included in the analysis. DATA EXTRACTION: Two reviewers independently reviewed the eligibility and abstracted data regarding the sample population, technical imaging characteristics of FDG-PET, and the number of true positives, true negatives, false positives and false negatives. Differences between readers were resolved by consensus. DATA SYNTHESIS: We used meta-analytic methods to estimate the pooled sensitivity, false positive rate, and the maximum joint sensitivity and specificity. Eighteen studies met the inclusion criteria. Sixteen studies included patient-based data, comprising a sample size of 808 subjects, and eight studies included lesion-based data, totaling 1013 lesions. Among the studies with patient-based data, the median sensitivity was 92.7%, and the median specificity was 81.6%. The pooled sensitivity was 90% [95% confidence interval (86.8-93.2)], and the pooled false positive rate was 11% [95% confidence interval (7.8-14.6)], after the exclusion of outliers. The maximum joint sensitivity and specificity, was 88% [95% confidence interval (86.0-90.6)]. CONCLUSION: These results indicate that FDG-PET is a valuable tool for detecting breast cancer recurrence and metastases. PMID- 15803357 TI - Interleukin-10 promoter polymorphism is associated with decreased breast cancer risk. AB - Interleukin-10 (IL-10) is an immunosuppressive cytokine which may facilitate development of cancer by supporting tumor escape from the immune response. A [TCATA] haplotype formed by polymorphisms at positions -3575, -2763, -1082, -819 and -592 in the promoter of the IL-10 gene is a strong determinant for IL-10 expression. The presence of this haplotype can be determined by analysis of the 592C > A polymorphism. Aim of the present study was to analyze the role of the IL 10 [TCATA] haplotype for breast cancer. We performed a case-control study including 500 female patients with histologically confirmed breast cancer and 500 female, age-matched, healthy control subjects from population-based screening studies. The -592C > A polymorphism was determined by a 5'-nuclease assay (TaqMan). Frequency of the homozygous -592 AA genotype, indicating homozygosity for the [TCATA] haplotype, was 4.2% among patients and 7.3% among controls (p=0.038; odds ratio 0.56; 95% confidence interval 0.32-0.97). IL-10 genotypes were not associated with tumor size, histological grading, estrogen or progesterone receptor status and age at diagnosis. Therefore we conclude that the IL-10 -592C > A promoter polymorphism may be associated with a reduced breast cancer risk. PMID- 15803358 TI - Clodronate treatment influences MMP-2 associated outcome in node positive breast cancer. AB - PURPOSE: Serum postoperative matrix metalloproteinase 2 (MMP-2) level is a predictor of outcome in node positive breast cancer and can be used to stratify patients into low and high risk groups. Our aim was to determine how clodronate treatment influences MMP-2 associated clinical outcome. PATIENTS AND METHODS: Women with primary node-positive breast cancer were randomized to control group or to receive oral clodronate for 3 years. Adjuvant chemo- or endocrine therapy was given to all patients. The follow-up time for all patients was 5 years. MMP-2 and MMP-9 levels were quantitatively measured from the serum of 252 patients before and after 1 year clodronate treatment using enzyme-linked immunoassays. RESULTS: In clodronate-treated patients, postoperative MMP-2 levels did not predict 5-year disease-free survival or overall survival (DFS, in low MMP-2 group (<5.32 ng/ml, median) 53% versus in high MMP-2 group (>median) 63%, p=NS; OS, 68% versus 63%, p=NS). When the patients were grouped according to serum MMP-2 levels, survival rates among patients with low MMP-levels were better in control than clodronate treated patients (DFS, 82% versus 53%, p = 0.003; OS, 91% versus 68%, p=0.014). Among patients with high serum MMP-2 levels, no significant difference in DFS or OS was found between control and clodronate groups. In multivariate analysis of low risk patients, independent predictors for DFS were treatment, age, nodal and PgR status, and those for OS treatment together with nodal and ER status. During 12 months follow-up, MMP-2 levels increased significantly more in clodonate receiving patients than in controls (p = 0.002). In comparison, when the patients were grouped according to MMP-9 levels, clodronate also impaired DFS among patients with low MMP-9 levels (82% versus 53%, p = 0.02), but no influence on OS was observed (83% versus 70%, p = 0.09). CONCLUSIONS: Clodronate interferes with the prognostic value of serum MMP-2. Clodronate has a negative impact on outcome among patients with low serum MMP-2 and MMP-9 levels, while no such influence is observed among patients with high MMP-2 and MMP-9 levels. PMID- 15803360 TI - Complementary and alternative medicine use is associated with an increased perception of breast cancer risk and death. AB - PURPOSE: Complementary and alternative medicines (CAMS) are frequently used by patients with breast cancer and their use may be related to the presence of psychosocial distress. The objective of this study is to assess the presence of anxiety and depression in breast cancer patients who use CAM and assess how they perceive their risks of recurrence and dying from breast cancer compared to non users. METHODS: Breast cancer patients attending ambulatory clinics at a single cancer center were approached by their oncologist. Participants completed a self administered survey regarding CAM usage, beliefs associated with CAM, views of their risks of developing recurrent cancer, and of dying of breast cancer. The presence of anxiety and depression was scored using the Hospital Anxiety and Depression Scale (HADS). Responses were scored and compared between CAM users and non-users. RESULTS: A total of 251 patients completed the survey. CAM usage was reported by 43% of patients. Individuals who used CAM were younger [53.1 versus 63.0 year (p < 0.001)], had higher education (p < 0.001), were more likely to have full time employment [25.7 versus 16.3% (p < 0.001)] and have private insurance for medications (76 versus 60%, p = 0.007) compared to non-users. CAM users perceived their risk of recurrence (p = 0.011) and death from breast cancer (p = 0.0001) as being significantly greater than non-users. There were no significant differences in anxiety or depression scores between CAM and non users. CONCLUSION: CAM use is associated with an increased perception of breast cancer recurrence and of breast cancer-related death. There was no association between the presence of anxiety or depression and CAM use. Improved patient understanding of actual risks of recurrence and death are required such that women will be able to make more informed decisions about using CAMs. PMID- 15803359 TI - Distinct breast cancer incidence and prognostic patterns in the NCI's SEER program: suggesting a possible link between etiology and outcome. AB - BACKGROUND: Breast cancer is a heterogeneous and chronic disease with relapses and death occurring 25 years or more after primary diagnosis. Standard tumor characteristics are used to predict initial relapse or death, but their ability to estimate long-term patterns of failure may be limited. METHODS: To further evaluate the significance of standard tumor features, we compared incidence and prognostic patterns in the National Cancer Institute's (NCI's) large-scale population-based Surveillance, Epidemiology, and End Results (SEER) program for high-risk versus low-risk breast cancers, i.e., size > 2.0 versus < or = 2.0 cm, lymph node positive versus negative, high versus low histologic grade, and hormone receptor negative versus positive expression, respectively. Data were stratified by age 50 years to approximate menopause. RESULTS: High-risk versus low-risk breast cancers demonstrated two very different incidence and prognostic patterns. Age-specific incidence rates among women with high-risk tumors increased until age 50 years then flattened, whereas rates among women with low risk tumors increased continuously with aging. Hazard rates for breast cancer death spiked sharply two years following primary breast cancer diagnosis among women with high-risk but not with low-risk tumors. Paradoxically, hazard function crossed over 6-8 years following breast cancer diagnosis, with hazard rates lower for high-risk than for low-risk breast cancers. CONCLUSION: Distinct incidence and prognostic patterns among high-risk and low-risk breast cancers suggest a possible link between breast cancer etiology and outcome. These epidemiologic results appear to complement emerging molecular genetic techniques, showing distinct genotypes for high-risk and low-risk breast cancer phenotypes. PMID- 15803361 TI - Genetic polymorphisms of TGF-beta1 & TNF-beta and breast cancer risk. AB - OBJECTIVE: The proliferation of malignant breast epithelial cells is regulated by various stimuli including cytokines and growth factors, thus the variants of those genes may modify the breast cancer risk. To evaluate the potential influences of TGF-beta1 T29C and TNF-beta A252G gene polymorphisms on breast cancer risk, a case-control study was conducted in Korea. METHODS: Histologically confirmed breast cancer cases (n=560) and controls (n=509) with no previous history of cancer were recruited from three teaching hospitals in Seoul, Korea. Genotypes were determined by PCR-CTPP (polymerase chain reaction with confronting two-pair primers) method. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by unconditional logistic regression model adjusting for age, body mass index, education, parity, age at first full-term pregnancy, and family history of breast cancer. RESULTS: The TGF-beta1 29C-allele containing genotypes posed an increased risk of breast cancer (OR=1.3, 95% CI=1.02-1.79), especially in postmenopausal women (OR=1.6, 95% CI=1.01-2.44). Similarly, the TNF-beta 252G allele containing genotypes posed an increased risk of postmenopausal breast cancer (OR=1.7, 95% CI=1.09-2.55). The risk of postmenopausal breast cancer increased in parallel with the number of the risk genotypes (p for trend <0.01). When data were stratified by the presumed non-genetic risk factors, TGF-beta1 C allele containing genotypes were found to increase breast cancer risk almost two fold in postmenopausal women with greater than median body mass index (>22.8 kg/m2) (OR=1.9, 95% CI=1.04-3.37). CONCLUSION: The results of this study therefore suggest that polymorphisms of TGF-beta1 and TNF-beta genes may modify individual susceptibility to breast cancer in Korean women. PMID- 15803362 TI - A phase II trial of imatinib mesylate monotherapy in patients with metastatic breast cancer. AB - BACKGROUND: Imatinib mesylate is a potent inhibitor of Abl, KIT, and PDGFR tyrosine kinases. Breast cancer has variable expression of KIT and PDGFR therefore we conducted a phase II trial to evaluate the safety and efficacy of imatinib in patients with metastatic breast cancer (MBC). PATIENTS AND METHODS: Eligible patients had measurable and progressive MBC, with no limits on prior chemo- or hormonal therapy. Imatinib was initially administered at a dose of 400 mg orally twice a day with provisions for dose reductions based on toxicities. The primary endpoint was clinical benefit based on RECIST criteria. Tumor specimens were tested for expression of KIT and PDGFR tyrosine kinases. RESULTS: Sixteen patients were enrolled and treated. Median age was 55 years (range: 35 73); median number of prior chemotherapy regimens for MBC was 4 (range 1-8). The main non-hematologic toxicities were (Grades 1/2; Grade 3): fatigue (56%; 6%), edema (38%; 19%), nausea (31%; 19%), vomiting (38%; 0%), anorexia (38%; 0%), diarrhea (19%; 6%), and rash (25%; 6%). Grade 3/4 hematologic and biochemical abnormalities were minimal. There was no evidence of clinical benefit. The median duration of therapy on trial was 28 days (range 2-71). Of the 13 testable cases: 1 was KIT positive and 4 were PDGFR positive. CONCLUSION: Imatinib therapy at doses of 800 mg/day was associated with significant toxicity in patients with heavily pre-treated MBC. Our results do not indicate activity for imatinib monotherapy in these unselected patients. PMID- 15803363 TI - The CHEK2 c.1100delC germline mutation rarely contributes to breast cancer development in the Czech Republic. AB - In this study we performed the CHEK2 c.1100delC mutation analysis in 1046 breast cancer patients and 730 unaffected control individuals. The mutated allele was found in 3 out of 688 unselected sporadic breast cancer patients and in 1 out of 358 familial/early onset breast cancer patients. Two mutations were identified in a cohort of 730 controls. Our results support the finding that frequency of CHEK2 c.1100delC mutation varies among different populations. Based on our results, genotyping of CHEK2 c.1100delC mutation in clinical settings in the Czech Republic could not be recommended. PMID- 15803364 TI - Primary docetaxel chemotherapy in patients with breast cancer: impact on response and survival. AB - Primary chemotherapy achieves high clinical response rates and facilitates breast conservation in many patients with large and locally advanced breast cancer. It may also serve to indicate responsiveness to chemotherapeutic agents. A pathological complete response to primary chemotherapy is a primary predictor and surrogate marker of long-term outcome, but occurs in only approximately 15% of patients. Docetaxel is of particular interest in this setting. Primary docetaxel chemotherapy has single-agent activity in both dose-dense and traditional schedules, with acceptable tolerability. Furthermore, concomitant docetaxel anthracycline schedules have shown promise in Phase II trials, achieving clinical overall response rates (ORRs) of 77-96%, pathological complete responses of up to 24%, and breast conservation in up to 89% of patients. Two Phase III studies have shown that pathological complete response is significantly improved with the addition of docetaxel to anthracycline-based therapy versus the latter alone: the Aberdeen study achieved a rate of 34% versus 16%, respectively (p = 0.04), and the NSABP-B27 study a rate of 26% versus 14%, respectively (p < 0.001). The Aberdeen study has suggested that the addition of docetaxel may yield a survival benefit at 5 years (p = 0.04). The Phase II GEPAR-DUO study hints at a benefit for sequential over concomitant docetaxel-based therapy, with improvements in both clinical response (ORR 87% versus 77%, respectively) and pathological complete response (23% versus 12%, respectively). Non-anthracycline docetaxel based primary regimens have shown early promise. As we continue to define the optimal regimen, a growing body of evidence supports the use of docetaxel in primary chemotherapeutic regimens for breast cancer. PMID- 15803365 TI - Pathology of breast cancer in women with constitutional CHEK2 mutations. AB - The power of association studies between polymorphic genetic variants and breast cancer may be enhanced if the cancer subjects are subclassified by histologic subgroup. In this study we classified 482 unselected breast cancers from Szczecin, Poland by histology (ductal, lobular, medullary, other). All women were genotyped for three founder mutations in the CHEK2 gene (1100delC, IVS2 + 1G > A and I157T). There was no significant overall association between CHEK2 and breast cancer (OR = 1.3; p = 0.30), but among those with lobular carcinoma the association with the I157T missense mutation was very strong (OR = 6.6; p > 0.0001). This is the first report to demonstrate that different mutations of the same gene may be associated with specific histologic subtypes of cancer. PMID- 15803366 TI - Is stereotactic large-core needle biopsy beneficial prior to surgical treatment in BI-RADS 5 lesions? PMID- 15803369 TI - Micromolar concentrations of 2-methoxyestradiol kill glioma cells by an apoptotic mechanism, without destroying their microtubule cytoskeleton. AB - The purpose of this study was to investigate the potential effects of 2 methoxyestradiol, a natural mammalian steroid, in glioma cells, since antiproliferative effects of this compound had been shown earlier in several leukemia and carcinoma cell lines. The effects of 0.2, 2 and 20 microM concentrations of 2-methoxyestradiol were measured in three malignant human glioma cell lines (U87MG, U138MG, LN405) and one malignant rat glioma cell line (RG-2) using a microtiter-tetrazolium (MTT) assay. In all cell lines, a significant reduction of the viable cell number by more then 75% occurred ( P < 0.05) for concentrations of 2 and 20 microM 2-methoxyestradiol after 6 days. A concentration of 0.2 microM had smaller effects (10-40% cell reduction), which were significant in two of the cell lines tested. The apoptotic nature of cell death was further analyzed in U87MG and RG-2 cells. Caspase-3 activity was significantly induced to levels between 3.4- and 23-fold after 4 days for the two higher 2-methoxyestradiol concentrations (P < 0.05). In the cell line RG-2 nuclear fragmentation was visible in many nuclei, following stains with Hoechst H33258. A round cell morphology occurred in most treated cells, which was not accompanied by a complete destruction of the microtubule network, as it can be observed with other microtubule targeting drugs. PMID- 15803368 TI - Inhibition of cell invasion by indomethacin on glioma cell lines: in vitro study. AB - Malignant glioma invasion into the surrounding brain tissue is still a major problem for any therapeutical methods. Matrix metalloproteinases (MMPs) have been implicated as important factors in this pathological process. In this study, one of the non-steroidal anti-inflammatory drugs (NSAIDs) indomethacin was employed to investigate the effect of inhibition of cell invasion mediated by MMP-2 and MMP-9 in human malignant glioma cell lines, A172, U87MG, U251MG, and U373MG in vitro. MTT assay was firstly examined to determine non-cytotoxic dose range, then gelatin zymography, matrigel invasion assay, migration assay and MMP-2 activity assay for 24 h exposure in indomethacin were employed to assess the inhibitory effect of indomethacin. MTT assay revealed that dose with 0, 50, and 500 microM/ml were non-cytotoxic. Zymography demonstrated: (a) expression of MMP-2 and MMP-9 activity was downregulated along with elevated dose of indomethacin. (b) MMP-2 activity that changed from pro-MMP-2 to active form of MMP-2 in supernatants of cell lines could not be inhibited by indomethacin. Invasion assay disclosed that the number of invading cells through the matrigel were significantly decreased in a dose dependent manner. Migration assay indicated indomethacin did not affect cells migration. MMP-2 activity assay showed the total and active MMP-2 secretion was suppressed by 500 microM/ml of indomethacin. Our present study is the first report on inhibitive effect of indomethacin mediated by MMP-2 and MMP-9 in invasion assay of glioma cell lines. The current study suggested that non-cytotoxic level of indomethacin was able to reduce the cell invasion of malignant gliomas mediated by MMP-2 and MMP-9, but it did not affected on cell motility. It also lowered down the activity of MMP-2 and MMP-9, and could reduce of MMP-2 secretion of cell lines. Thus, high concentration of indomethacin within non-cytotoxic dose might offer a new therapeutic strategy to impair cell invasion of gliomas. PMID- 15803370 TI - Antisense bcl-2 transfection up-regulates anti-apoptotic and anti-oxidant thioredoxin in neuroblastoma cells. AB - Antisense bcl-2 therapy combined with chemotherapy has been proved to be effective in various tumors. However, the role played by antisense bcl-2 therapy alone is not clear. In this study, we compared the apoptosis and the protein profiles of antisense bcl-2 transfected human neuroblastoma SK-N-MC cells to the control cells. Flow cytometric data indicated that antisense bcl-2 transfection did not lead to more extensive apoptosis in SK-N-MC cells (14.9 +/- 3.8%) than the control cells (10.3 +/- 2.3%). The above observation was confirmed by fluorescence microscopy using Hoechst 33258 staining. However, antisense bcl-2 induced changes in the expression of various proteins as shown by proteomic comparison, which included the up-regulation of the anti-apoptotic and anti oxidant protein thioredoxin. By western blot validation, thioredoxin was found to be up-regulated by 2.9-folds with the corresponding down-regulation of Bcl-2 by 2.1-folds. The up-regulation of thioredoxin may be a compensating mechanism for cell survival in neuroblastoma when Bcl-2 expression is suppressed, and it may to some extent attenuate the effectiveness of antisense bcl-2 therapy. PMID- 15803371 TI - Analysis of chromosome 7 in adult and pediatric ependymomas using chromogenic in situ hybridization. AB - Few studies have yielded reliable data that distinguish between ependymal neoplasms based on molecular genetic attributes. The present study utilizes chromogenic in situ hybridization (CISH), a relatively recent hybridization technique, to retrospectively examine chromosome 7-copy number in pediatric and adult ependymomas. Of the 27 hybridizations, polysomy of chromosome 7 was detected in 10 out of 15 (66%) adult ependymomas, and in only three out of 12 (25%) pediatric lesions. All myxopapillary ependymomas showed polysomy. The remaining tumors were diploid. The authors conclude that (1) there are distinct genetic subsets of ependymoma, in particular, increases in copy number of chromosome 7 are almost exclusively found in myxopapillary ependymoma, and that (2) CISH is a rapid and sensitive method of stratifying morphological variants of ependymoma and potentially other central nervous system (CNS) tumors. These results encourage further investigations with CISH on a larger scale to determine its merit as an ancillary diagnostic and prognostic tool. PMID- 15803372 TI - DeltaNp73 antisense activates PUMA and induces apoptosis in neuroblastoma cells. AB - The p73 gene codes for various different protein isoforms. They include proteins expressed under the control of the P1 promoter that contain a transactivation domain and are similar in function to p53 (TAp73 isoforms), as well as proteins regulated by the P2 promoter that lack this domain and function as dominant negative inhibitors of TAp73 and p53 (DeltaNp73 isoforms). Whereas TAp73 functions as a tumor suppressor with pro-apoptotic function, DeltaNp73 is likely to prevent the induction of apoptosis in tumor cells and to participate in oncogenesis. Here we used a loss-of-function strategy to assess the role of DeltaNp73 in SH-SY5Y neuroblastoma cells. An antisense oligonucleotide designed to target DeltaNp73 mRNA, but not TAp73, was used to effectively downregulate this transcript. DeltaNp73 downregulation was accompanied by increased levels of the pro-apoptotic BH3 family member PUMA at the mRNA and protein level, and by conformational activation of BAX which translocated to mitochondria. These DeltaNp73 antisense-mediated alterations led to the induction of apoptosis as detected by decreased cell viability, augmented DNA fragmentation and increased caspase-3 activity in cell lysates. Our results demonstrate the cytoprotective role of DeltaNp73 in neuroblastoma and suggest its use as a target for molecular intervention therapy. PMID- 15803374 TI - Optic nerve head Drusen mimicking papilledema. PMID- 15803373 TI - Analysis of target genes induced by IL-13 cytotoxin in human glioblastoma cells. AB - IL-13 cytotoxin comprised of IL-13 and a mutated form of Pseudomonas exotoxin (fusion protein termed IL-13-PE38QQR) has been shown to inhibit protein synthesis leading to necrotic and apoptotic cell death in glioblastoma cells that express high levels of interleukin-13 receptors (IL-13R). To identify target genes of cell death and other cellular genes with IL-13 receptors in glioblastoma cells, we utilized the cDNA microarrays to analyze global gene expression profiles after IL-13 cytotoxin and IL-13 treatment. IL-13 cytotoxin mediated cytotoxicity to U251 cells in a dose-dependent manner. Hierarchical cluster analysis of differentially expressed genes in U251 glioma cells at different time points after IL-13 cytotoxin treatment showed three major groups, each representing a specific expression pattern. Randomly selected differentially expressed genes from each group were confirmed by RT-PCR analysis. Most down-regulated genes belong to cell adhesion, motility, angiogenesis, DNA repair, and metabolic pathways. While up-regulated genes belong to cell cycle arrest, apoptosis, signaling and various metabolic pathways. Unexpectedly, at early time points, both IL-13 and IL-13 cytotoxin induced several genes belonging to different pathways most notably IL-8, DIO2, END1, and ALDH1A3 indicating that these genes are early response genes and their products may be associated with IL-13R. In addition, IL-13 cytotoxin induced IL-13Ralpha2 mRNA expression during the treatment in glioma cells. Our results indicate that novel cellular genes are involved with IL-13 receptors and that IL-13 cytotoxin induced cell death involves various target genes in human glioblastoma cells. On going studies will determine the role of associated genes and their products in the IL-13R functions in glioma cells. PMID- 15803375 TI - Paragangliomas in the cauda equina region: clinicopathoradiologic findings in four cases. AB - Paragangliomas of the cauda equina are rare neuroendocrine tumors. Four cases of nonsecreting paraganglioma of the cauda equina, preoperatively misdiagnosed as neurinoma, are presented with an emphasis on the correlation between magnetic resonance imaging findings and pathological features. Although it is difficult to correctly diagnose paraganglioma preoperatively for intradural extramedullary tumors, especially in the cauda equina, paraganglioma should be included in differential diagnoses. PMID- 15803376 TI - Human interferon beta, nimustine hydrochloride, and radiation therapy in the treatment of newly diagnosed malignant astrocytomas. AB - Previous investigators have reported encouraging results for malignant gliomas treated using a combination of human interferon beta (IFN-beta) with nimustine hydrochloride (ACNU) and radiation therapy (termed IAR therapy). This study was undertaken to examine further the efficacy of the IAR regimen followed by maintenance therapy with IFN-beta and ACNU in patients with newly diagnosed malignant astrocytomas. Fifty-eight patients were enrolled onto the trial. IFN beta (2 x 10(6) IU/m(2)/day x 5 days/week for 8 consecutive weeks) and ACNU (80 mg/m(2) on days 1 and 36) were administered intravenously concomitant with radiation therapy followed by IFN-beta (every 2 weeks) and ACNU (every 6 weeks). Of 33 patients assessable for a response, 11 responded (33%), with 4 complete responses. The estimated median overall survival (OS) was 16 months, with values of 58 and 13 months for anaplastic astrocytoma (AA) and glioblastoma (GB) patients, respectively. The overall progression free survival (PFS) was 11 months, with values of 31 and 7 months for AA and GB patients, respectively. The IAR therapy was safe and well tolerated. Based on a statistical analysis of the factors that affected the PFS and OS, histologic grade, postoperative Karnofsky performance scale (KPS), and extent of surgery were identified as independent predictors. The postoperative KPS stood out as the most powerful prognostic factor, which was also the best predictor for the response to IAR therapy. Our findings suggest a possible benefit for IAR therapy followed by maintenance therapy mainly in AA. In addition, they emphasize the importance of a preserved KPS after cytoreductive surgery, which could produce a potential benefit for adjuvant therapy and could ultimately lead to a prolonged survival. PMID- 15803377 TI - Paraneoplastic recurrent multifocal encephalitis presenting with epilepsia partialis continua. AB - We report a 46-year-old female patient in whom epilepsia partialis continua was the initial presentation of small cell lung cancer. Magnetic resonance imaging revealed multiple, bilateral cortical lesions, which were originally misinterpreted and treated as brain metastases. Intracranial lesions and neurological symptoms remitted after corticosteroids, chemotherapy, and radiotherapy. After an asymptomatic interval of 18 months, neurological symptoms recurred with more extensive involvement of completely different parts of the central nervous system (limbic, brainstem and cerebellar structures) without any evidence of tumor recurrence. Both episodes showed a distinctive response to immunosuppressive therapy. The diagnostic challenges of the highly variable clinical presentations and therapeutic approaches to paraneoplastic multifocal encephalitis are discussed with relevant literature review. PMID- 15803378 TI - The treatment of advanced sinonasal malignancies with pre-operative intra arterial cisplatin and concurrent radiation. AB - INTRODUCTION: Malignancies of the nasal and paranasal sinuses are uncommon tumors, accounting for only 3% of all aerodigestive tract neoplasms. Despite advances in surgical techniques and continued evolution of adjuvant therapies, the 5-year mortality remains unusually high at greater than 50%. In 1996, we begin utilizing a novel strategy in the treatment of advanced sinonasal carcinomas. This consisted of neoadjuvant selective intra-arterial cisplatin with concurrent radiation therapy (acronym RADPLAT) followed by a conservative craniofacial resection. We now report our results for 11 patients treated with this regimen. METHODS: Between July 1996 and April 2003, 11 patients with advanced sinonasal malignancies underwent treatment utilizing the RADPLAT protocol followed by a planned surgical resection via a craniofacial approach. Patient charts, operative notes, follow-up clinic notes, and pre- and post operative imaging studies were reviewed in detail for each subject. RESULTS: Histopathological analysis of the tumors revealed seven squamous cell carcinomas (64%), two adenocarcinomas (18%), one adenoid cystic carcinoma (9%), and one sinonasal undifferentiated carcinoma (9%). T4N0M0 disease was present in nine patients (81%), and two patients had T3N0M0 disease (19%). Survival was calculated using the Kaplan-Meier method with an overall survival of 81% at 5 years and a progression-free survival at 5 years of 67%. Mean follow-up is 57.2 months ranging from 12 to 95 months. CONCLUSIONS: The treatment of advanced sinonasal malignancies with pre-operative intra-arterial cisplatin and concurrent radiation results in a significant improvement in survival. This can be done safely with high response rates and excellent loco-regional control in T3 and T4 disease. Although are results are encouraging, there is a need for a cooperative, multi-institutional, prospective study. PMID- 15803380 TI - Atypical teratoid/rhabdoid tumor case report: treatment with surgical excision, radiation therapy, and alternative medicines. AB - Intracranial atypical teratoid/rhabdoid tumors (AT/RT) are rare with a poor prognosis. We report one case of a 7-year old girl living over 17 months after the diagnosis of AT/RT in the left frontal lobe. Treatment was partial surgical resection and post-operative radiation therapy. Radiation therapy resulted in complete response with no evidence of residual or recurrent disease more than 17 months after diagnosis. The patient has been maintained on an extensive regimen of alternative therapies since completion of radiation therapy. PMID- 15803379 TI - Continuous remission of newly diagnosed and relapsed central nervous system atypical teratoid/rhabdoid tumor. AB - Atypical teratoid/rhabdoid tumors (AT/RT) are highly malignant lesions of childhood that carry a very poor prognosis. AT/RT can occur in the central nervous system (CNS AT/RT) and disease in this location carries an even worse prognosis with a median survival of 7 months. In spite of multiple treatment regimens consisting of maximal surgical resection (including second look surgery), radiation therapy (focal and craniospinal), and multi-agent intravenous, oral and intrathecal chemotherapy, with or without high-dose therapy and stem cell rescue, only seven long-term survivors of CNS AT/RT have been reported, all in patients with newly diagnosed disease. For this reason, many centers now direct such patients, particularly those under 5 years of age, or those with recurrent disease, towards comfort care rather than attempt curative therapy. We now report on four children, two with newly diagnosed CNS AT/RT and two with progressive disease after multi-agent chemotherapy who are long term survivors (median follow-up of 37 months) using a combination of surgery, radiation therapy, and intensive chemotherapy. The chemotherapy component was modified from the Intergroup Rhabdomyosarcoma Study Group (IRS III) parameningeal protocol as three of the seven reported survivors in the literature were treated using this type of therapy. Our four patients, when added to the three reported survivors in the literature using this approach, suggest that patients provided this aggressive therapy can significantly alter the course of their disease. More importantly, we report on the first two survivors after relapse with multi-agent intravenous and intrathecal chemotherapy treated with this modified regimen. PMID- 15803381 TI - Glioneuronal tumor with unique imaging and histologic features. PMID- 15803382 TI - The many and varied niches occupied by Yersinia pestis as an arthropod-vectored zoonotic pathogen. AB - Yersinia pestis, the causative agent of bubonic and pneumonic plague, has a complex lifestyle, cycling between both arthropod and mammalian hosts. This pathogen has previously been shown to survive intracellularly within macrophages and to be capable of biofilm formation within the flea, suggesting the development of a range of strategies to ensure survival throughout its life cycle, including expression of virulence factors and tight regulation of its genes. PMID- 15803383 TI - Rhizobacteria and their potential to control Fusarium verticillioides: effect of maize bacterisation and inoculum density. AB - Fusarium verticillioides is the most important seed transmitted pathogen that infects maize. It produces fumonisins, toxins that have potential toxicity for humans and animals. Control of F. verticillioides colonisation and systemic contamination of maize has become a priority area in food safety research. The aims of this research were (1) to characterise the maize endorhizosphere and rhizoplane inhabitant bacteria and Fusarium spp., (2) to select bacterial strains with impact on F. verticillioides growth and fumonisin B1 production in vitro, (3) to examine the effects of bacterial inoculum levels on F. verticillioides root colonisation under greenhouse conditions. Arthrobacter spp. and Azotobacter spp. were the predominant genera isolated from maize endorhizosphere and rhizoplane at the first sampling period, whilst F. verticillioides strains showed the greatest counts at the same isolation period. All F. verticillioides strains were able to produce fumonisin B1 in maize cultures. Arthrobacter globiformis RC5 and Azotobacter armeniacus RC2, used alone or in a mix, demonstrated important effects on F. verticillioides growth and fumonisin B1 suppression in vitro. Only Azotobacter armeniacus RC2 significantly reduced the F. verticillioides root colonisation at 10(6) and 10(7) CFU g(-1) levels under greenhouse conditions. PMID- 15803384 TI - Streptomyces nanningensis sp. nov. a novel Streptomycete from forest soil. AB - A strain YIM 33098T (= CCTCC AA001027T = DSM 41831T) was isolated from a forest soil sample collected from Nanning in Guangxi Province, China, in the course of screening for producers of new drug lead compounds. This strain was identified by using a polyphasic approach. The results showed that it should be assigned to the genus Streptomyces. An almost complete 16S rRNA gene sequence of the strain was determined and compared with those of representative Streptomyces species. Strain YIM 33098T was clustered in the same subclade with Streptomyces tendae ATCC19812T and Streptomyces eurythermus ATCC14975T. Similarities of strain YIM 33098T with the two strains were 97.35% and 97.42%, respectively. Based on the phenotypic and genotypic evidence, it is therefore proposed that strain YIM 33098T should be classified in the genus Streptomyces as a new species under the name of Streptomyces nanningensis sp. nov. PMID- 15803385 TI - Aspergillus vadensis, a new species of the group of black Aspergilli. AB - A strain from the group of black Aspergilli was analysed in detail to determine the species to which it belongs. A detailed analysis of morphology, RFLP patterns and metabolite profiles was carried out. In addition, a phylogenetic tree was constructed for the black Aspergilli using the ITS and the beta-tubulin sequences of the individual strains. The new species differs by its poor growth on glycerol and galacturonate and its unique extrolite profile consisting of aurasperone B, nigragillin, asperazine and kotanins. RFLP analysis using three genes as probes also resulted in a unique pattern. These data indicate that the strain was closely related but not identical to Aspergillus foetidus, Aspergillus niger and Aspergillus tubingensis. It was therefore designated as a novel species and named Aspergillus vadensis. PMID- 15803386 TI - Molecular cloning and functional characterization of fumarases C in Neisseria species. AB - Fumarase is one of the key enzymes in the TCA cycle and has been implicated in virulence and survival of some microorganisms under suboptimal environmental conditions. In this study, the fumC genes that encode fumarase C (FUMCs) from Neisseria meningitidis, N. gonorrhoeae and N. subflava were identified by homology-based analysis, cloned by polymerase chain reactions and fully sequenced. The inferred primary sequence of neisserial FUMCs showed a high degree of conservation with 97.8-98.7% amino acid identity. However, phylogenetic analysis revealed that these neisserial FUMCs are divergent from class II fumarases found in other microorganisms, rat and human. The putative fumC genes were subcloned into the expression vector, pGEX-6P-1 and efficiently expressed in Esherichia coli BL21. The purified recombinant fusion proteins obtained by affinity chromatography demonstrated high catalytic activities (120-180 U/mg), thus authenticating the identities and functionalities of the cloned genes. Whether FUMC has any physiological relevance to the pathogenesisity of Neisseriae must await future gene disruption or mutagenesis studies. PMID- 15803387 TI - Streptomyces roseoalbus sp. nov., an actinomycete isolated from soil in Yunnan, China. AB - An actinomycete strain was isolated from a soil sample collected from a secondary forest at Yongsheng of Yunnan province, China. The isolate, YIM 31634T, was identified by a polyphasic approach. The 16S rDNA sequence analysis showed that the strain YIM 31634T belongs to the genus Streptomyces, with closest similarity to Streptomyces olivochromogenes DSM 40451T (97.66% similarity). Sequence similarities between strain YIM 31634T and other Streptomyces species in the same subclade ranged from 97.59% (with Streptomyces resistomycificus DSM 40133T) to 97.22% (with Streptomyces mirabilis ATCC 27447T). Key phenotypic characteristics as well as chemotaxonomic features of the actinomyces were congruent with the description of the genus Streptomyces. On the basis of phenotypic and phylogenetic analyses, strain YIM31634T was recognized as a new species of the genus Streptomyces for which the name Streptomyces roseoalbus sp. nov. is proposed. The strain YIM 31634T has been deposited in the Chinese Center of Type Culture Collection as strain CCTCC M 203016T and in the Deutsche Sammlung von Mikroorganismen (DSM 41833T). PMID- 15803388 TI - Characterisation of the nitrile hydratase gene clusters of Rhodococcus erythropolis strains AJ270 and AJ300 and Microbacterium sp. AJ115 indicates horizontal gene transfer and reveals an insertion of IS1166. AB - The nitrile metabolising strains AJ270, AJ300 and AJ115 were isolated from the same location. The strains have very similar nitrile metabolising profiles. Sequencing of the 16S rRNA gene indicates that strains AJ270 and AJ300 are novel strains of Rhodococcus erythropolis while strain AJ115 is a novel Microbacterium strain very closely related to Microbacterium oxydans and Microbacterium liquefaciens. Analysis of the structure of the nitrile hydratase/amidase gene clusters in the three strains indicates that this region is identical in these strains and that this structure is different to other nitrile hydratase/amidase gene clusters. The major difference seen is the insertion of a complete copy of the insertion sequence IS1166 in the nhr2 gene. This copy of IS1166 generates a 10 bp direct duplication at the point of insertion and has one ORF encoding a protein of 434 amino acids, with 98% homology to the transposase of IS666 from Mycobacterium avium. A gene oxd, encoding aldoxime dehydratase is found upstream of the nitrile hydratase gene cluster and an open reading frame encoding a protein with homology to GlnQ type ABC transporters is found downstream of the nitrile hydratase/amidase genes. The identity of the nitrile hydratase/amidase gene clusters in the three strains suggests horizontal gene transfer of this region. Analysis of the strains for both linear and circular plasmids indicates that both are present in the strains but hybridisation studies indicate that the nitrile hydratase/amidase gene cluster is chromosomally located. The nitrile hydratase/amidase enzymes of strain AJ270 are inducible with acetonitrile or acetamide. Interestingly although a number of Fe-type nitrile hydratases have been shown to be photosensitive, the enzyme from strain AJ270 is not. PMID- 15803389 TI - Characterization of the AXDH gene and the encoded xylitol dehydrogenase from the dimorphic yeast Arxula adeninivorans. AB - The xylitol dehydrogenase-encoding Arxula adeninivorans AXDH gene was isolated and characterized. The gene includes a coding sequence of 1107 bp encoding a putative 368 amino acid protein of 40.3 kDa. The identity of the gene was confirmed by a high degree of homology of the derived amino acid sequence to that of xylitol dehydrogenases from different sources. The gene activity was regulated by carbon source. In media supplemented with xylitol, D-sorbitol and D-xylose induction of the AXDH gene and intracellular accumulation of the encoded xylitol dehydrogenase was observed. This activation pattern was confirmed by analysis of AXDH promoter-GFP gene fusions. The enzyme characteristics were analysed from isolates of native strains as well as from those of recombinant strains expressing the AXDH gene under control of the strong A. adeninivorans-derived TEF1 promoter. For both proteins, a molecular mass of ca. 80 kDa was determined corresponding to a dimeric structure, an optimum pH at 7.5 and a temperature optimum at 35 degrees C. The enzyme oxidizes polyols like xylitol and D-sorbitol whereas the reduction reaction is preferred when providing D-xylulose, D-ribulose and L-sorbose as substrates. Enzyme activity exclusively depends on NAD+ or NADH as coenzymes. PMID- 15803390 TI - Cloning and expression of a lipoxygenase from Pseudomonas aeruginosa 42A2. AB - In order to produce (S) 10-monohydroxy-8E-octadecenoic acid (MHOD) from oleic acid, a full-length probable lipoxygenase cDNA from Pseudomonas aeruginosa 42A2 was cloned and expressed in Escherichia coli BL21(DE3). The recombinant protein was purified by affinity chromatography to electrophoretic homogeneity and specifically stained. Its molecular mass was 70 kDa. The activity of the rec-LOX with oleic acid was about 30% of that of the preferred substrate, linoleic acid (100%). Bacterial LOX forms a new subfamily in the lipoxygenase phylogenetic tree. PMID- 15803391 TI - Transcription analysis of pilS and xpsEL genes from Xylella fastidiosa. AB - Xylella fastidiosa is a xylem-limited phytopathogen responsible for diseases in several plants such as citrus and coffee. Analysis of the bacterial genome revealed some putative pathogenicity-related genes that could help to elucidate the molecular mechanisms of plant-pathogen interactions. In the present work, the transcription of three genes of the bacterium, grown in defined and rich media and also in media containing host plant extracts (sweet orange, 'ponkan' and coffee) was analyzed by RT-PCR. The pilS gene, which encodes a sensor histidine kinase responsible for the biosynthesis of fimbriae, was transcribed when the bacterium was grown in more complex media such as PW and in medium containing plant extracts. The xps genes (xpsL and xpsE) which are related to the type II secretion system were also detected when the bacterium was grown in rich media and media with 'ponkan' and coffee extracts. It was thus observed that pilS and xpsEL genes of X. fastidiosa can be modulated by environmental factors and their expression is dependent on the nutritional status of the growth medium. PMID- 15803393 TI - Structure of calmodulin complexed with an olfactory CNG channel fragment and role of the central linker: residual dipolar couplings to evaluate calmodulin binding modes outside the kinase family. AB - The NMR high-resolution structure of calmodulin complexed with a fragment of the olfactory cyclic-nucleotide gated channel is described. This structure shows features that are unique for this complex, including an active role of the linker connecting the N- and C-lobes of calmodulin upon binding of the peptide. Such linker is not only involved in the formation of an hydrophobic pocket to accommodate a bulky peptide residue, but it also provides a positively charged region complementary to a negative charge of the target. This complex of calmodulin with a target not belonging to the kinase family was used to test the residual dipolar coupling (RDC) approach for the determination of calmodulin binding modes to peptides. Although the complex here characterized belongs to the (1--14) family, high Q values were obtained with all the 1:1 complexes for which crystalline structures are available. Reduction of the RDC data set used for the correlation analysis to structured regions of the complex allowed a clear identification of the binding mode. Excluded regions comprise calcium binding loops and loops connecting the EF-hand motifs. PMID- 15803395 TI - 1H-15N correlation spectroscopy of nanocrystalline proteins. AB - The limits of resolution that can be obtained in 1H-15N 2D NMR spectroscopy of isotopically enriched nanocrystalline proteins are explored. Combinations of frequency switched Lee-Goldburg (FSLG) decoupling, fast magic angle sample spinning (MAS), and isotopic dilution via deuteration are investigated as methods for narrowing the amide 1H resonances. Heteronuclear decoupling of 15N from the 1H resonances is also studied. Using human ubiquitin as a model system, the best resolution is most easily obtained with uniformly 2H and 15N enriched protein where the amides have been exchanged in normal water, MAS at approximately 20 kHz, and WALTZ-16 decoupling of the 15N nuclei. The combination of these techniques results in average 1H lines of only approximately 0.26 ppm full width at half maximum. Techniques for optimizing instrument stability and 15N decoupling are described for achieving the best possible performance in these experiments. PMID- 15803394 TI - Measurement of eight scalar and dipolar couplings for methine-methylene pairs in proteins and nucleic acids. AB - A new 3D, spin-state-selective coherence transfer NMR experiment is described that yields accurate measurements for eight scalar or dipolar couplings within a spin system composed of a methylene adjacent to a methine group. Implementations of the experiment have been optimized for proteins and for nucleic acids. The experiments are demonstrated for Cbeta-Calpha moieties of the third IgG-binding domain from Streptococcal Protein G (GB3) and for C5'-C4' groups in a 24-nt RNA oligomer. Chemical shifts of Calpha, Cbeta and Hbeta (respectively C4', C5' and H5') are dispersed in the three orthogonal dimensions, and the absence of heteronuclear decoupling leads to distinct and well-resolved E.COSY multiplet patterns. In an isotropic sample, the E.COSY displacements correspond to 1J(CalphaHalpha), 2J(CalphaHbeta2)+2J(CalphaHbeta3), 2J(CbetaHalpha), 1J(CbetaHbeta2)+1J(CbetaHbeta3), 1J(CbetaHbeta2)-2J(Hbeta2Hbeta3), 1J(CbetaHbeta3)-2J(Hbeta2Hbeta3), 3J(HalphaHbeta2) and 3J(HalphaHbeta3) for proteins, and 1J(C4'H4'), 2J(C4'H5')+2J(C4'H5"), 2J(C5'H4'), 1J(C5'H5')+1J(C5'H5"), 1J(C5'H5')-2J(H5'H5"), 1J(C5'H5")-2J(H5'H5"), 3J(H4'H5') and 3J(H4'H5") in nucleic acids. The experiment, based on relaxation-optimized spectroscopy, yields best results when applied to residues where the methine methylene group corresponds to a reasonably isolated spin system, as applies for C, F, Y, W, D, N and H residues in proteins, or the C5'-C4' groups in nucleic acids. Splittings can be measured under either isotropic or weakly aligned conditions, yielding valuable structural information both through the 3J couplings and the one-, two- and three-bond dipolar interactions. Dipolar couplings for 10 out of 13 sidechains in GB3 are found to be in excellent agreement with its X-ray structure, whereas one residue adopts a different backbone geometry, and two residues are subject to extensive chi1 rotamer averaging. The abundance of dipolar couplings can also yield stereospecific assignments of the non-equivalent methylene protons. For the RNA oligomer, dipolar data yielded stereospecific assignments for six out of the eight C5'H2 groups in the loop region of the oligomer, in all cases confirmed by 1J(C5'H5')>1J(C5'H5"), and H5' resonating downfield of H5". PMID- 15803398 TI - NMR assignment of the R-module from the Azotobacter vinelandii Mannuronan C5 epimerase AlgE4. PMID- 15803396 TI - Accurate measurement of 15N-13C residual dipolar couplings in nucleic acids. AB - New 3D HCN quantitative J (QJ) pulse schemes are presented for the precise and accurate measurement of one-bond 15N1/9-13C1', 15N1/9-13C6/8, and 15N1/9-13C2/4 residual dipolar couplings (RDCs) in weakly aligned nucleic acids. The methods employ 1H-13C multiple quantum (MQ) coherence or TROSY-type pulse sequences for optimal resolution and sensitivity. RDCs are obtained from the intensity ratio of H1'-C1'-N1/9 (MQ-HCN-QJ) or H6/8-C6/8-N1/9 (TROSY-HCN-QJ) correlations in two interleaved 3D NMR spectra, with dephasing intervals of zero (reference spectrum) and approximately 1/(2J(NC)) (attenuated spectrum). The different types of 15N 13C couplings can be obtained by using either the 3D MQ-HCN-QJ or TROSY-HCN-QJ pulse scheme, with the appropriate setting of the duration of the constant-time 15N evolution period and the offset of two frequency-selective 13C pulses. The methods are demonstrated for a uniformly 13C, 15N-enriched 24-nucleotide stem loop RNA sequence, helix-35psi, aligned in the magnetic field using phage Pf1. For measurements of RDCs systematic errors are found to be negligible, and experiments performed on a 1.5 mM helix-35psi sample result in an estimated precision of ca. 0.07 Hz for 1D(NC), indicating the utility of the measured RDCs in structure validation and refinement. Indeed, for a complete set of 15N1/9 13C1', 15N1/9-13C6/8, and 15N1/9-13C2/4 dipolar couplings obtained for the stem nucleotides, the measured RDCs are in excellent agreement with those predicted for an NMR structure of helix-35psi, refined using independently measured observables, including 13C-1H, 13C-13C and 1H-1H dipolar couplings. PMID- 15803397 TI - Determining the optimal size of small molecule mixtures for high throughput NMR screening. AB - High-throughput screening (HTS) using NMR spectroscopy has become a common component of the drug discovery effort and is widely used throughout the pharmaceutical industry. NMR provides additional information about the nature of small molecule-protein interactions compared to traditional HTS methods. In order to achieve comparable efficiency, small molecules are often screened as mixtures in NMR-based assays. Nevertheless, an analysis of the efficiency of mixtures and a corresponding determination of the optimum mixture size (OMS) that minimizes the amount of material and instrumentation time required for an NMR screen has been lacking. A model for calculating OMS based on the application of the hypergeometric distribution function to determine the probability of a "hit" for various mixture sizes and hit rates is presented. An alternative method for the deconvolution of large screening mixtures is also discussed. These methods have been applied in a high-throughput NMR screening assay using a small, directed library. PMID- 15803399 TI - 1H, 13C, and 15N resonance assignments of SAP18. PMID- 15803400 TI - 1H, 13C, and 15N assignments of MMP-12, a key protease implicated in lung tissue remodeling. PMID- 15803401 TI - NMR assignment of the Xenopus laevis prion protein fragment xlPrP (98-226). PMID- 15803402 TI - Backbone and side-chain 1H, 15N, and 13C assignments for the beta domain of the bacterial cell division protein DivIB. PMID- 15803403 TI - 1H, 15N, and 13C resonance assignments of reduced glutaredoxin C1 from Populus tremula x tremuloides. PMID- 15803404 TI - 1H, 15N and 13C assignments of the alkaline proteinase inhibitor APRin from Pseudomonas aeruginosa. PMID- 15803406 TI - 1H, 15N and 13C backbone and side-chain assignments of the rice phytochrome B PAS1 domain and backbone assignments of the PAS1-PAS2 domain. PMID- 15803405 TI - Resonance assignments and secondary structureof hPrxVI, a 25 kDa 1-cys human peroxiredoxin enzyme. PMID- 15803407 TI - Backbone and side chain resonance assignmentsof a TRAV14-3 mouse T cell receptor domain. PMID- 15803408 TI - Gene cloning and characterization of a soybean (Glycine max L.) LEA protein, GmPM16. AB - Late embryogenesis abundant (LEA) proteins, present in abundance in seeds during the late stages of development, are associated with desiccation tolerance. In the present work, we characterize a soybean LEA protein, GmPM16, with low molecular weight, high pI value, and an unusual amino acid residue distribution along the protein. The transcripts were detected in cotyledon mesophyll cells but not in the vascular system of mature or pod-dried soybean seeds. Circular dichroism (CD) analysis and Fourier transfer infrared (FTIR) spectroscopy indicated that the GmPM16 protein in solution was highly unordered, possessing only partial alpha helical structures. However, the protein in sodium dodecyl sulfate (SDS) or trifluoroethanol (TFE) solution or in a dry state exhibited a conformation of abundant alpha-helical structures. As well, the GmPM16 protein interacts with sugar and forms tightly glassy matrixes in the dry state. The protein may play a role in reducing cellular damage in drying seeds by changing the protein conformation and forming tight cellular glasses. PMID- 15803409 TI - Allelic characterization of the leaf-variegated mutation var2 identifies the conserved amino acid residues of FtsH that are important for ATP hydrolysis and proteolysis. AB - Arabidopsis var1 and var2 mutants exhibit leaf variegation. VAR1 and VAR2 encode similar FtsH metalloproteases (FtsH5 and FtsH2, respectively). We have previously found many variegated mutants to be allelic to var2. Each mutant was shown to express a different degree of variegation, and the formation of white sectors was enhanced in severely variegated alleles when these alleles were grown at low temperature. VAR1/FtsH5 and VAR2/FtsH2 levels were mutually affected even in the weak alleles, confirming our previous observation that the two proteins form a hetero complex. In this study, the sites of the mutations in these var2 alleles were determined. We isolated eight point mutations. Five alleles resulted in an amino acid substitution. Three of the five amino acid substitutions occurred in Walker A and B motifs of the ATP-binding site, and one occurred in the central pore motif. These mutations were considered to profoundly suppress the ATPase and protease activities. In contrast, one mutation was found in a region that contained no obvious signature motifs, but a neighboring sequence, Gly-Ala-Asp, was highly conserved among the members of the AAA protein family. Site-directed mutagenesis of the corresponding residue in E. coli FtsH indeed showed that this residue is necessary for proper ATP hydrolysis and proteolysis. Based on these results, we propose that the conserved Gly-Ala-Asp motif plays an important role in FtsH activity. Thus, characterization of the var2 alleles could help to identify the physiologically important domain of FtsH. PMID- 15803410 TI - Altered expression of an ankyrin-repeat protein results in leaf abnormalities, necrotic lesions, and the elaboration of a systemic signal. AB - The PR-like proteins, class I beta-1,3-glucanase (GLU I) and chitinase (CHN I), are induced as part of a stereotypic response that can provide protection against viral, bacterial, and fungal pathogens. We have identified two Nicotiana plumbaginifolia ankyrin-repeat proteins, designated Glucanohydrolase Binding Proteins (GBP) 1 and 2, that bind GLU I and CHN I both in vitro and when expressed in yeast cells. Sense as well as antisense transformants of tobacco carrying the GBP1 gene elaborated graft-transmissible acropetally moving signals that induced the downward curling of young leaves. This phenotype was associated with reduced starch, sucrose, and fructose accumulation; the formation of necrotic lesions; and, the induction of markers for the hypersensitive response. GBP1/2 are members of a conserved Plant- Specific Ankyrin- repeat (PANK) family that includes proteins implicated in carbohydrate allocation, reactive oxygen metabolism, hypersensitive cell death, rapid elicitor responses, virus pathogenesis, and auxin signaling. The similarity in phenotype of PANK transformants and transformants altered in carbohydrate metabolism leads us to propose that PANK family members are multifunctional proteins involved in linking plant defense responses and carbohydrate metabolism. PMID- 15803411 TI - A gymnosperm ABI3 gene functions in a severe abscisic acid-insensitive mutant of Arabidopsis (abi3-6) to restore the wild-type phenotype and demonstrates a strong synergistic effect with sugar in the inhibition of post-germinative growth. AB - The CnABI3 gene of yellow-cedar is an orthologue of the ABI3/VP1 gene of angiosperms; it shares many common characteristics with other ABI3/VP1 genes, yet has unique characteristics as well. We examined whether this gymnosperm transcription factor can functionally complement an angiosperm species with a defective ABI3 gene. A severe Arabidopsis abi3 null mutant abi3-6 was stably transformed with the CnABI3 gene coding-region driven by a modified CaMV 35S promoter. Several of the visible mutant phenotypes (e.g., production of green seeds due to a lack of chlorophyll breakdown) were fully restored to those of the wild-type and the transformed seeds acquired desiccation tolerance. The functional complementation of the mutant also extended to the accumulation of several seed proteins (including seed-storage-proteins, alpha-tonoplast intrinsic protein, dehydrin-related polypeptides and oleosin), which were restored to wild type levels. However, not all phenotypes were fully restored; sensitivities of transgenic seeds to exogenous ABA (as far as germination is concerned) were lower than that of the wild-type seeds, and flowering times were intermediate of those characteristic of wild-type and abi3-6 plants. A novel function for CnABI3, potentially related to a direct or indirect role in ER homeostasis was revealed. Two proteins with a molecular chaperone function in the ER (BiP and protein disulphide isomerase) were elevated in mutant seeds (indicative of ER stress); expression of the CnABI3 gene decreased the accumulation of these proteins to levels characteristic of the wild-type. These studies reveal the degree of conservation of ABI3 functions between gymnosperms and angiosperms as well as some novel functions of ABI3-related genes. PMID- 15803412 TI - AKINbeta3, a plant specific SnRK1 protein, is lacking domains present in yeast and mammals non-catalytic beta-subunits. AB - The SNF1/AMPK/SnRK1 heterotrimeric kinase complex is involved in the adaptation of cellular metabolism in response to diverse stresses in yeast, mammals and plants. Following a model proposed in yeast, the kinase targets are likely to bind the complex via the non-catalytic beta-subunits. These proteins currently identified in yeast, mammals and plants present a common structure with two conserved interacting domains named Kinase Interacting Sequence (KIS) and Association with SNF1 Complex (ASC), and a highly variable N-terminal domain. In this paper we describe the characterisation of AKINbeta3, a novel protein related to AKINbeta subunits of Arabidopsis thaliana, containing a truncated KIS domain and no N-terminal extension. Interestingly the missing region of the KIS domain corresponds to the glycogen-binding domain (beta-GBD) identified in the mammalian AMPKbeta1. In spite of its unusual features, AKINbeta3 complements the yeast sip1Deltasip2Deltagal83Delta mutant. Moreover, interactions between AKINbeta3 and other AKIN complex subunits from A. thaliana were detected by two-hybrid experiments and in vitro binding assays. Taken together these data demonstrate that AKINbeta3 is a beta-type subunit. A search for beta-type subunits revealed the existence of beta3-type proteins in other plant species. Furthermore, we suggest that the AKINbeta3-type subunits could be plant specific since no related sequences have been found in any of the other completely sequenced genomes. These data suggest the existence of novel SnRK1 complexes including AKINbeta3-type subunits, involved in several functions among which some could be plant specific. PMID- 15803414 TI - The Arabidopsis KLUNKER gene controls cell shape changes and encodes the AtSRA1 homolog. AB - The analysis of a group of seven trichome mutants in Arabidopsis, which all show distorted trichomes along with severe actin defects has revealed insight into the role of the actin cytoskeleton in cell shape control. Four of the corresponding genes encode components of a protein complex, the ARP2/3 complex that stimulates the production of 'fine actin' at active growth sites. In this study, we show that another member of the distorted group, KLUNKER (KLK), encodes the AtSRA1 homolog of Arabidopsis and that klk mutants show a similar range of cell shape defects to those of arp2/3 mutants. In animals, SRA1 regulates the activity of the ARP2/3-regulating WAVE-HSPC300 complex in a Rho-dependent manner. Our findings provide evidence that a Rho/ARP2/3 regulation pathway exists in plants. PMID- 15803413 TI - The Fed-1 (CAUU)4 element is a 5' UTR dark-responsive mRNA instability element that functions independently of dark-induced polyribosome dissociation. AB - Darkness rapidly induces a decline in the stability and translation of the pea Ferredoxin-1 (Fed-1) mRNA in transgenic tobacco. Direct half-life measurement showed that mutation of the (CAUU)4 stabilizes Fed-1 mRNA in the dark. (CAUU)1, a feature more common in plant 5' UTRs than (CAUU)4, confers slight light responsive mRNA accumulation. At least three but less than 11 CAUU repeats near the 5' end of the 5' UTR are required for full light-responsive accumulation. Furthermore, 26 nt of the 5' UTR, including the (CAUU)4 repeat, is sufficient to confer a significant approximately 2.5-fold increase in light-regulated mRNA accumulation when fused to the 5' end of a heterologous plant mRNA. A mutation of the (CAUU)4 repeat that compromises light-regulated mRNA stability changes in vitro the accessibility of the region to ribonuclease V1 and ribonuclease A suggesting the geometry formed by the repeat may be important for instability. Finally, dark-induced Fed-1 mRNA instability occurs even when most of the mRNA is retained on polyribosomes, and thus is likely an independent event regulated by darkness. PMID- 15803415 TI - Toward a blueprint for UDP-glucose pyrophosphorylase structure/function properties: homology-modeling analyses. AB - UDP-glucose pyrophosphorylase (UGPase) is an important enzyme of synthesis of sucrose, cellulose, and several other polysaccharides in all plants. The protein is evolutionarily conserved among eukaryotes, but has little relation, aside from its catalytic reaction, to UGPases of prokaryotic origin. Using protein homology modeling strategy, 3D structures for barley, poplar, and Arabidopsis UGPases have been derived, based on recently published crystal structure of human UDP-N acetylglucosamine pyrophosphorylase. The derived 3D structures correspond to a bowl-shaped protein with the active site at a central groove, and a C-terminal domain that includes a loop (I-loop) possibly involved in dimerization. Data on a plethora of earlier described UGPase mutants from a variety of eukaryotic organisms have been revisited, and we have, in most cases, verified the role of each mutation in enzyme catalysis/regulation/structural integrity. We have also found that one of two alternatively spliced forms of poplar UGPase has a very short I-loop, suggesting differences in oligomerization ability of the two isozymes. The derivation of the structural model for plant UGPase should serve as a useful blueprint for further function/structure studies on this protein. PMID- 15803416 TI - Characterization of the genomic structures and selective expression profiles of nine class I small heat shock protein genes clustered on two chromosomes in rice (Oryza sativa L.). AB - The cytosolic class I small heat shock proteins (sHSP-CI) represent the most abundant sHSP in plants. Here, we report the characterization and the expression profile of nine members of the sHSP-CI gene family in rice (Oryza sativa Tainung No.67), of which Oshsp16.9A, Oshsp16.9B, Oshsp16.9C, Oshsp16.9D and Oshsp17.9B are clustered on chromosome 1, and Oshsp17.3, Oshsp17.7, Oshsp17.9A and Oshsp18.0 are clustered on chromosome 3. Oshsp17.3 and Oshsp18.0 are linked by a 356-bp putative bi-directional promoter. Individual gene products were identified from the protein subunits of a heat shock complex (HSC) and from in vitro transcription/ translation products by two-dimensional gel electrophoreses (2 DE). All sHSP-CI genes except Oshsp17.9B were induced strongly after a 2-h heat shock treatment. The genes on chromosome 3 were induced rapidly at 32 and 41 degrees C, whereas those on chromosome 1 were induced slowly by similar conditions. Seven of these genes, except Oshsp16.9D and Oshsp17.9B, were induced by arsenite (As), but only genes on chromosome 3 were strongly induced by azetidine-2-carboxylic acid (Aze, a proline analog) and cadmium (Cd). A similar expression profile of all sHSP-CI genes at a lower level was evoked by ethanol, H2O2 and CuCl2 treatments. Transient expression assays of the promoter activity by linking to GUS reporter gene also supported the in vivo selective expression of the sHSP-CI genes by Aze treatment indicating the differential induction of rice sHSP-CI genes is most likely regulated at the transcriptional level. Only Oshsp16.9A abundantly accumulated in mature dry seed also suggested additionally prominent roles played by this HSP in development. PMID- 15803417 TI - A fragment of chloroplast DNA was transferred horizontally, probably from non eudicots, to mitochondrial genome of Phaseolus. AB - The mitochondrial genomes of some Phaseolus species contain a fragment of chloroplast trnA gene intron, named pvs-trnA for its location within the Phaseolus vulgaris sterility sequence (pvs). The purpose of this study was to determine the type of transfer (intracellular or horizontal) that gave rise to pvs-trnA. Using a PCR approach we could not find the respective portion of the trnA gene as a part of pvs outside the Phaseolus genus. However, a BLAST search revealed longer fragments of trnA present in the mitochondrial genomes of some Citrus species, Helianthus annuus and Zea mays. Basing on the identity or near identity between these mitochondrial sequences and their chloroplast counterparts we concluded that they had relocated from chloroplasts to mitochondria via recent, independent, intracellular DNA transfers. In contrast, pvs-trnA displayed a relatively higher sequence divergence when compared with its chloroplast counterpart from Phaseolus vulgaris. Alignment of pvs-trnA with corresponding trnA fragments from 35 plant species as well as phylogenetic analysis revealed that pvs-trnA grouped with non-eudicot sequences and was well separated from all Fabales sequences. In conclusion, we propose that pvs-trnA arose via horizontal transfer of a trnA intron fragment from chloroplast of a non-eudicot plant to Phaseolus mitochondria. This is the first example of horizontal transfer of a chloroplast sequence to the mitochondrial genome in higher plants. PMID- 15803420 TI - Dry eye--is this a disease or a natural consequence of aging? PMID- 15803422 TI - Diagnostic criteria in dry eye. PMID- 15803418 TI - Functional conservation and maintenance of expression pattern of FIDDLEHEAD-like genes in Arabidopsis and Antirrhinum. AB - In Arabidopsis, loss of function of the epidermis-specific FDH gene coding for a putative beta-ketoacyl-CoA synthase results in ectopic organ fusions in mutants. Corresponding mutants are not available for Antirrhinum majus, however, organ fusions can be induced in both species by chloroacetamide inhibitors of beta ketoacyl-CoA synthases using a chemical genetics approach. We isolated the ortholog of FDH from Antirrhinum majus, the ANTIRRHINUM FIDDLEHEAD (AFI ) gene, and showed that AFI complements fdh when expressed in the epidermis under control of the FDH promoter. Like FDH, the AFI gene exhibits protodermis- and epidermis specific expression, and its promoter directs the expression of reporter genes to the epidermis in transgenic Antirrhinum and Arabidopsis. We demonstrate down regulation of the FDH promoter in the epidermis of the ovary septum, thereby supporting the assumption that FDH-like genes may directly facilitate the cell cell interactions that need to occur during carpel fusion and pollen tube growth. Up-regulation of FDH in the stomium, on the other hand, provides evidence for its possible involvement in cell separation during anther dehiscence. Down-regulation of the FDH and AFI promoters in the septum is observed in transgenic Arabidopsis but not in Antirrhinum plants. This probably reflects differences in the ontogeny of the ovary septum between the two species. We also show that epidermis-specific FDH-like genes may not be able to efficiently elongate fatty acid chains when misexpressed in seeds. PMID- 15803424 TI - [Drug induced uveitis]. AB - PURPOSE: To review the world literature about drug induced uveitis. METHODS: We used the OMS and Naranjo criteria of causality of adverse drug reactions to classify the causes of drug induced uveitis. These have been classified into certain, probable and possible causes. We described the clinical features associated with each drug. RESULTS: Certain causes of uveitis are byphosphonates, topiramato and metipranolol eyedrops. Probable causes are rifabutin, sulfonamides, cidofovir, immunorecovery in patients following HAART therapy, topical corticosteroids, latanoprost, and intravitreous triamcinolone acetate. Possible causes of uveitis include bimatoprost, travoprost, brimonidine, other eyedrops and intraocular drugs, tubulointerstitial nephritis and uveitis (TINU) produced by nonsteroidal drugs, iopamidol, diethylcarbamazepine, interleukins 3 and 6, oral contraceptives, quinidine, vaccines and cobalt, ibuprofen and moxifloxacin. CONCLUSION: Drug induced uveitis is an entity of low incidence, anterior iridocyclitis is the commonest clinical picture, visual acuity is moderately diminished, and if use of the drug is ceased uveitis does not recur. PMID- 15803425 TI - [Nomogram for ocular hypertension progression risk based on the ocular hypertension treatment study]. AB - INTRODUCTION: A practical nomogram has been designed in order to present the results obtained from the Ocular Hypertension Study (Gordon et al. Arch Ophthalmol 2002; 120: 714-720), where the relation between intraocular pressure (IOP) and corneal thickness becomes apparent, involving the risk of evolution from ocular hypertension into glaucoma within a 6 year period. MATERIAL AND METHODS: We used a multiple logarithmic regression for the nine parameters shown in figure 1 of the above mentioned paper. RESULTS: A correlation coefficient of 0.91 (p<0.001) permits to establish the following equation: Probability of evolution (%) = 13539.5 x (1.1385IOP) x (0.9818(CORNEAL THICKNESS)). This implies that a variation of 10 microns on corneal thickness leads to an IOP's modification of 1.5 mmHg in the same sense. From these data, we designed the nomogram included in this paper. CONCLUSIONS: IOP and pachymetry together allow an estimation of the risk of evolution from ocular hypertension into glaucoma in a graphical practical way. From this indirect estimation, the influence of corneal thickness on IOP's measure seems to be much higher than previously estimated. PMID- 15803426 TI - [Surgical treatment of third cranial nerve palsy]. AB - PURPOSE: To study surgical treatment results of third cranial nerve palsy. METHOD: We have retrospectively studied 11 patients with a third cranial nerve palsy (10 unilateral and 1 bilateral) who were subjected to surgery. The etiology was traumatic in 4 cases, congenital in 2, vascular in 4, and tumoral in 1. Torticollis was present in 7 patients and 5 had diplopia. A good aesthetic result was defined by a final deviation of less than 10 prismatic diopters, a medium result by a deviation between 10-20 diopters and a bad result if the deviation was > or = 20 diopters. A good functional result was considered if there was no diplopia when the eyes were in their primary position. RESULTS: In all instances large recessions and/or resections of horizontal recti muscles were performed. Vertical deviation was treated in 4 cases. Multiple surgery was needed in 3 cases, 2 patients being operated on twice and 1 case was operated on four times. Botulinum toxin was used in 4 cases, before or after surgery. The aesthetic result was good in 7 cases, medium in 2, and bad in the other 2. Postoperative diplopia was present in 4 cases. Mean postoperative follow-up was 27.9 months. CONCLUSIONS: Large recti muscle recessions and resections are our first surgical technique to treat problems of third cranial nerve palsy. Sometimes several surgeries are required to treat vertical deviation and previous hypocorrection. Botulinum toxin is a complementary treatment. Functional results are worse than aesthetic ones. PMID- 15803427 TI - [Atypical presentation of ocular hypertension following myopic LASIK]. AB - OBJECTIVE: We present a masquerade syndrome recently described following uneventful laser in situ keratomileusis (LASIK). If misdiagnosed or left unattended, this condition can lead to severe visual loss. METHODS: Findings concerning ten patients who underwent uneventful myopic LASIK, but who presented with a granular haze in the interface in the early postoperative period, are described. Initially, some of the patients were treated for diffuse lamellar keratitis with an intensive course of topical steroids, but, rather than improving, the haziness of the interface worsened or remained the same, and was even accompanied by the development of epithelial edema. RESULTS: Tonometry led to the diagnosis of ocular hypertension induced by the steroid therapy. The clinical picture resolved completely with topical hypotensive treatment and with the cessation of steroid treatment. CONCLUSION: In the early postoperative period following LASIK an atypical presentation of ocular hypertension can occasionally be observed. A high index of suspicion of this diagnosis must be maintained due to the inaccuracy of tonometry after LASIK and the similarity of this clinical picture with diffuse lamellar keratitis. These cases emphasize the importance of taking into account underestimation of the intraocular pressure after myopic LASIK. PMID- 15803428 TI - [Transpupillary thermotherapy in the primary management of choroidal melanoma]. AB - OBJECTIVE: To determine if primary transpupillary thermotherapy can achieve local remission of choroidal melanoma. Secondary objectives were to determine safety of treatment, visual acuity and evaluate complications. The long term objective was to evaluate the impact of treatment on survival without metastatic disease. METHODS AND PATIENTS: Thirty-one patients with small choroidal melanomas were treated with transpupillary thermotherapy between 1st December 1997 and 15th June 2001. These patients satisfied predetermined inclusion criteria. To perform the treatment, an infrared diode laser (810 nm) with emission adapted for use with a surgical microscope was used, applying the treatment through a Mainster contact lens. In most patients, the diameter of the irradiation beam was 2 mm, the exposure time was 1 minute and the level of energy given was between 250 and 800 mW. RESULTS: Of 31 patients treated, 26 patients were evaluated in this study. With a mean follow-up of 23.13 months (standard deviation (S.D.): 15 and median: 18.6 months), tumoral remission was observed in 50% of the evaluated tumors. The post-treatment visual acuity in the patients with tumoral remission was better than in patients with treatment failure (P = 0.004). CONCLUSIONS: Primary treatment of choroidal melanomas with transpupillary thermotherapy may be a good therapeutic option in selected cases. PMID- 15803429 TI - [Evaluation of refractive prognosis factors in myopic LASIK]. AB - PURPOSE: Our objective was to evaluate myopic Lasik refractive results according to preoperative refraction findings. METHODS: In 200 eyes, classified into 4 groups according to preoperative sphere and cylinder findings, different postoperative refractive parameters were compared: predictability, visual acuity (VA), security index (SI) and efficacy index (EI). RESULTS: Grouped by sphere: spherical correction was better than cylindrical correction. Highest negative spheres resulted in undercorrection (p<0.0171). The residual astigmatism was similar and the predictability within +/- 1.0 D was lower for the most myopic groups. Grouped according to cylinder: the higher the cylinder, the more cylinder undercorrection (p<0.05) and more sphere overcorrection was achieved (p=0.0469). Between the lowest and highest cylinder levels, the residual astigmatism difference was of -0.32 x 90 degrees and predictability +/-1.0 D was of 98 and 94% for the sphere and 96 and 88% for the cylinder respectively. Defocus Equivalent <1 D for the lowest and highest myopic groups was 74 vs. 46% for the cylinder and 82 vs. 38% for the sphere respectively. Pre and postoperative VA were lower for higher spheres or cylinders. SI >1.0 was similar to EI (+/-0.9), for preoperative sphere and cylinder groups. Best corrected visual acuity (BCVA) of 20/25 by sphere was 100% vs. 60% and of 20/30 by cylinder was 100% vs. 83%, for lower and higher myopic groups respectively. CONCLUSIONS: Analysis by grouping preoperative refractive errors is helpful for establishing its influence and improving LASIK prognosis and treatment. This procedure is effective and safe for all groups, and its inaccuracy is due more to astigmatism than to spherical error. As astigmatism increases, less spherical and higher cylindrical ablation should be performed. PMID- 15803430 TI - [Prevalence of diabetic retinopathy in the Regional Comunity of Extremadura. 1997 2001 (Extremadura Project to Prevent Blindness)]. AB - OBJECTIVE: To establish the prevalence of diabetic retinopathy (DR), types and risk factors in a diabetic population in Extremadura. METHODS: A cross-sectional study, with prospective data collection, was performed between 1997 and 2001 on 3,114 randomly selected diabetic patients belonging to 13 Diabetologic Assistance Centers. We evaluated: age, gender, type, treatment, duration of diabetes and blood pressure levels. Two color fundus photographs were taken and ophthalmoscopy was performed on both eyes. Data were analyzed using SPSS (11.5). RESULTS: The mean age of the patients was 63.8 (SD13.4) years, 7.5% were type 1 diabetics, 31.1% were insulin-treated and 46.0% had elevated blood pressure. Prevalence of DR was 35.7%, macular edema: 5.6% and proliferative DR: 5.3%. Multivariate analysis showed that type 1 diabetics have a DR frequency 1.79 times higher than type 2 diabetics (ORaj 1.79; IC 95% 1.22-2.63; p=0.003). Patients who have been diabetic for 5-10 years have a frequency 2 times higher (ORaj 2.00; IC 95% 1.62 2.45; p<0.001), patients diabetic for more than 15 years have a frequency 5.48 times higher (ORaj 5.48; IC 95% 4.33-6.93; p<0.001) than patients who have been diabetic for less than 5 years. The frequency of DR in patients aged more than 60 years is 23% higher than those under that age. (ORaj 1.23; IC 95% 1.01-1.50; p=0.045). CONCLUSIONS: 37.5% of the diabetic population in Extremadura is affected by diabetic retinopathy. New analyses are necessary in this population to detect the existence of other high risk subgroups in order to reduce these high prevalence levels. PMID- 15803431 TI - [The eye in the history of the human body composition (1556) (II)]. PMID- 15803433 TI - How do thiazide and thiazide-like diuretics lower blood pressure? AB - Thiazide diuretics are widely used for the treatment of hypertension, but the mechanism by which these drugs lower blood pressure in the long term remains unknown. This article reviews current knowledge about the hypotensive actions of thiazides and thiazide-like diuretics and discusses possible mechanisms of action. PMID- 15803434 TI - The revised role of ACE-inhibition after myocardial infarction in the thrombolytic/primary PCI era. AB - Many studies have investigated the process of left ventricular (LV) dilatation and the effects of angiotensin-converting enzyme (ACE) inhibitors after myocardial infarction (MI). It has been generally accepted that progression of LV dilatation is a major predictor of heart failure and death after MI. Also, attenuation of LV dilatation is thought to be one of the main mechanisms by which ACE inhibitors (ACE-Is) produce their beneficial effects. However, evidence for this hypothesis came from studies that were performed before thrombolytic therapy and primary percutaneous coronary intervention (PCI) were routinely used after acute MI. Nowadays, reperfusion is obtained much more frequently and LV dilatation after MI has become less prevalent. Nevertheless, ACE-Is proved effective in reducing cardiac morbidity and mortality. Therefore, mechanisms other than attenuation of LV dilatation, such as anti-atherosclerotic effects or plaque stabilisation, may explain the long-term beneficial effects of ACE-Is after MI. In the present overview, we evaluate the role of LV dilatation and the effects of ACE-Is after MI in the thrombolytic/primary PCI era and provide recommendations on ACE-I use in clinical practice. PMID- 15803435 TI - Angiotensin II stimulates arachidonic acid release from bone marrow stromal cells. AB - INTRODUCTION: Angiotensin II (Ang II) is recognised as a regulator of haematopoiesis, but its actions within the bone marrow are not fully understood. Support of haematopoiesis by bone marrow stromal cells (MSC) is dependent on factors that include arachidonic acid and macrophage colony stimulating factor (MCSF), both of which are increased by Ang II stimulation in other tissues. To further elucidate the mechanisms of Ang II-regulated haematopoiesis, we determined whether Ang II-stimulation alters arachidonic acid release and MCSF secretion from MSC. METHODS: Cynomolgus monkey MSC isolated from bone marrow aspirates and the human HS-5 stromal cell line were studied for Ang II-mediated arachidonic acid (AA) release, while secretion of MCSF in response to Ang II was studied in HS-5 cells. Cells were labelled overnight with 3H-AA and the release of 3H-AA was measured in culture medium following 20 minutes stimulation with Ang II, alone or in combination with the AT1- or AT2-receptor antagonists, losartan and PD 123319, respectively. MCSF secretion into culture medium was measured using an enzyme immunoassay following 24 hours of treatment with Ang II alone or in combination with losartan or PD 123319. Phorbol-myristate-acetate, known to stimulate release of AA and MCSF, was used as a positive control in both experiments. RESULTS: In response to Ang II, release of 3H-AA from monkey and human MSC was increased (p<0.05) to 147+/-4% and 124+/-3% of control, respectively. The AT1- and AT2-receptor antagonists, losartan and PD 123319, individually reduced Ang II-stimulated 3H-AA release. In contrast, Ang II had no effect on secretion of MCSF from HS-5 cells. CONCLUSIONS: These results provide mechanistic evidence for Ang II-mediated haematopoiesis through AA release that may, in part, explain Ang II-facilitated recovery of haematopoiesis in experimental myelosuppression and the anaemias associated with Ang II receptor blockade. PMID- 15803436 TI - Effect of bestatin on angiotensin I-, II- and III-induced collagen gel contraction in cardiac fibroblasts. AB - OBJECTIVE: The purpose of this investigation was to determine whether the aminopeptidase inhibitor with broad specificity, bestatin, affects angiotensin I (Ang I)-, angiotensin II (Ang II)- or angiotensin III (Ang III)-stimulated collagen gel contraction in cardiac fibroblasts. DESIGN AND METHODS: Cardiac fibroblasts (from normal male adult rats) were cultured to confluency in Dulbeccos modified Eagles medium (DMEM) with 10% foetal bovine serum (FBS). These fibroblasts (100,000 cells) were then further incubated in a floating collagen gel lattice with the test products Ang I (1 micromol/L), Ang II (100 nmol/L), Ang III (100 nmol/L) and bestatin (100 micromol/L) for three days in DMEM without FBS. The area of the collagen gels embedded with cardiac fibroblasts was determined by a densitometric analysis. Aminopeptidase activity was estimated by spectrophotometric determination of the liberation of p-nitroaniline from alanine or arginine-p-nitroanilide. RESULTS: Ang I, II and III stimulated (p<0.05) collagen gel contraction by 30.4+/-4.8 (SEM)%, 27.1+/-3.1% and 15.4+/-3.6% respectively. Ang I- and II-induced stimulation of collagen gel contraction was of the same order but more pronounced (p<0.05) than Ang III- stimulated collagen gel contraction. The Ang I-, II- and III-stimulated collagen contraction was reduced by bestatin. Bestatin, however, did not affect basal collagen gel contraction in cardiac fibroblasts. Bestatin dose-dependently inhibited the hydrolysis of arginine- and alanine-p-nitroanilide in cardiac fibroblasts. When a neutralising antibody to transforming growth factor TGF-b1 was added to the collagen gel simultaneously with the angiotensins, the stimulated collagen contraction was not affected. Beta-aminoproprionitrile, an inhibitor of lysyl oxidase, completely abolished basal as well as Ang I-, II- and III-stimulated collagen contraction in cardiac fibroblasts. RESULTS: Our data suggest that aminopeptidases are involved in the Ang I-, II- and III-induced stimulation of collagen contraction in cardiac fibroblasts. PMID- 15803437 TI - Lectin chromatography of extrarenal renin protein in human plasma and tissues: potential endocrine function via the renin receptor. AB - Secretion of prorenin from extrarenal tissues comprises approximately half of the renin protein in plasma; its origin is unknown. The discovery of a prorenin/renin receptor that activates vascular tissue kinases raises interest in this otherwise inactive component. We have studied its glycosylation as this may distinguish it from renal renin. The binding of renin protein (active and prorenin) in human plasma and tissues to concanavalin (Con A) and wheat germ lectins was deployed. Immunoradiometric and enzyme kinetic assays were applied to column fractions. Mannosylated renin protein binds to Con A and has been shown to be taken up by human vascular and hepatic cells on mannose-6-phosphate receptors, possibly as a clearance mechanism. But the other binding sites of prorenin/renin that elicit a cellular phosphorylation response are apparently independent of glycosylation. The tissues examined (kidney, adrenal, ovary) each contain high proportions of Con A binding renin, but the plasma of normal resting males and females contain mainly non-binding renin, the proportion increasing as renal renin secretion decreases. The extreme of this relationship is seen in anephric patients and in some normal women on oral contraceptives with suppressed renal renin, in whom plasma renin is entirely non-binding to Con A. Conversely, when renal renin secretion was stimulated, the increased plasma active renin bound to Con A. However, extrarenal tissues containing exclusively Con A non-binding renin protein, and hence potential sources of anephric plasma renin protein, were not identified, but are unlikely to include adrenal or female reproductive tract. The findings are consistent with the view that normal human plasma contains a considerable amount of amannosylated prorenin of extrarenal origin that escapes hepatic clearance and has a longer half-life than renal renin. This plasma renin form would be expected to be activated in association with the recently described renin/prorenin aspartyl protease receptor and to participate in local pathophysiological processes. PMID- 15803438 TI - Renal targeting of captopril using captopril-lysozyme conjugate enhances its antiproteinuric effect in adriamycin-induced nephrosis. AB - INTRODUCTION: High-sodium intake blunts the renoprotective efficacy of angiotensin-converting enzyme (ACE) inhibitors. We investigated whether targeting the drug to the kidneys may attenuate the inferior response to ACE inhibitor (ACE I) under high-sodium conditions. The ACE-I, captopril, was coupled to the low molecular weight protein (LMWP) lysozyme, yielding captopril-lysozyme conjugates that accumulate specifically in the proximal tubular cells of the kidneys. We compared the antiproteinuric efficacy of captopril to that of the captopril lysozyme conjugate in adriamycin-induced proteinuric rats fed with a high-sodium diet. MATERIALS AND METHODS: Rats with adriamycin (single injection 2 mg/kg)- induced proteinuria were put on a high-sodium diet (HS; 3% NaCl). When stable proteinuria developed at 5.5 weeks, animals were assigned to the following subcutaneous treatments: (1) vehicle (n=7); (2) lysozyme (equivalent to the amount in conjugate) (n=7); (3) captopril (5 mg/kg/24 hours) (n=8); (4) captopril lysozyme conjugate (captopril content equivalent to 1mg captopril/kg/24 hours) (n=7). Blood pressure and proteinuria were monitored. After 10 days of treatment the rats were sacrificed and kidneys and plasma were removed. RESULTS: Results are given as mean + S.E.M. After injection with adriamycin at t=0, stable proteinuria developed, amounting to 547+79 mg/24 hours at week 5.5. Subsequently, after seven and nine days of treatment, no reduction of proteinuria was observed in the captopril-treated group. In contrast, a significant reduction in proteinuria, amounting to 35+4% (day seven) and 25+2% (day nine), was observed in the captopril-lysozyme conjugate group (p<0.05 compared with the captopril group). In contrast, blood pressure was reduced in the captopril-treated group by 13.9+2.9 mmHg, while in the captopril-lysozyme treated group, an increase of 7.9+3.3 mmHg was found. Renal ACE activity was lowered by 30% in the captopril, as well as in the captopril-lysozyme conjugate treated group, compared with control. Furthermore, the ratio of kidney: plasma levels of captopril almost doubled as a consequence of coupling to lysozyme. CONCLUSION: In proteinuric rats fed with a high-sodium diet, captopril induced a reduction in blood pressure without an effect on proteinuria. In contrast, renal targeting of a five times lower dose of the ACE-I with the captopril-lysozyme conjugate reduced proteinuria without reducing blood pressure. Therefore, renal targeting of ACE-I may be a promising strategy to optimise the therapeutic response of ACE-I. PMID- 15803439 TI - Angiotensin (1-7) re-establishes impulse conduction in cardiac muscle during ischaemia-reperfusion. The role of the sodium pump. AB - INTRODUCTION: The effect of angiotensin (1-7) (Ang 1-7) on membrane potential and excitability of rat heart muscle under ischaemia/reperfusion was investigated. MATERIALS AND METHODS: The hearts of adult rats were removed under deep anaesthesia and perfused using the Langendorff method. After 40 minutes of global no-flow ischaemia, the heart was reperfused for five minutes and the right ventricle was dissected out and transferred to a transparent chamber, through which normal oxygenated Krebs solution flowed continuously (37 degrees C). Measurements of membrane potential were performed using an intracellular microelectrode connected to a high impedance preamplifier. The muscle was stimulated with rectangular current pulses (3 ms duration; 0.6 Hz) generated by an electronic stimulator and isolation unit. To study the influence of Ang (1-7) on sodium pump current, isolated myocytes were voltage-clamped at -40 mV and the current generated by the pump was recorded before and after the administration of Ang (1-7) (10-8 M) to the bath. RESULTS: Ang (1-7) (10-8 M) hyperpolarised the ischaemic heart fibre and re-established impulse propagation. The increment of resting potential was related to the activation of the sodium pump. Indeed, Ang (1-7) (10-8 M) enhanced the transient outward current generated by an electrogenic sodium pump. Both effects of Ang (1-7) on membrane potential and pump current were abolished by ouabain (10-7 M). The cardiac refractoriness was also increased by Ang (1-7) (10-8 M). CONCLUSIONS: Ang (1-7) activates the sodium pump, hyperpolarises the heart cell and re-establishes the impulse conduction during ischaemia/reperfusion. These effects of Ang (1-7), and the increment of cardiac refractoriness, provide an explanation for the reduced incidence of arrhythmias during ischaemia/reperfusion in the presence of Ang (1-7). PMID- 15803440 TI - A method to estimate the variance of an endpoint from an on-going blinded trial. AB - Blinded estimation of variance allows for changing the sample size without compromising the integrity of the trial. Some of the methods that estimate the variance in a blinded manner either make untenable assumptions or are only applicable to two-treatment trials. We propose a new method for continuous endpoints that makes minimal assumptions. The method uses the enrollment order of subjects and the randomization block size to estimate the variance. It can be applied to normal or non-normal data, trials with two or more treatments, equal or unequal allocation schemes, fixed or random randomization block sizes, and single or multi-centre trials. The variance estimator is unbiased and performs best when the randomization block size is the smallest. Simulation results suggest that for many commonly used randomization block sizes the proposed estimator is expected to perform well. The proposed method is used to estimate the variance of the endpoint for two trials and is shown to perform well by comparison with its unblinded counterpart. PMID- 15803441 TI - Sequential urn designs with elimination for comparing K > or =3 treatments. AB - A fully sequential procedure is proposed for comparing K > or =3 treatments with immediate binary responses. The procedure uses an adaptive urn design to randomize patients to the treatments and stopping rules are incorporated for eliminating less promising treatments. Simulation is used to assess the performance of the procedure for several adaptive urn designs, in terms of expected numbers of treatment failures and allocation proportions, and the effect on estimation at the end of the trial is also addressed. It is concluded that the drop-the-loser rule is more effective than equal allocation and all of the other designs considered. The practical benefits of the procedure are illustrated using the results of a three-treatment lung cancer study. It is then shown how the sequential elimination procedure may be used in dose-finding studies and its performance is compared with a recently proposed method. Several possible extensions to the work are briefly indicated. PMID- 15803442 TI - Cost-effectiveness analysis for multinational clinical trials. AB - Clinical trials of cost-effectiveness are often conducted in more than one country. The two most common ways of dealing with the multinational nature of the data are either to calculate a pooled estimate or to stratify results by country. Since the between-country heterogeneity in costs is potentially substantial, pooled estimates may be difficult to interpret for any one country. Policy decisions are often made at a national level, and so country-specific results are important. However, country-specific analyses will be based on fewer patients and will often fail to provide adequate precision for statistical analyses. Shrinkage estimation is a compromise between these two methods and has been used successfully in other fields. These estimates are country-specific yet less variable than those derived through a subgroup approach. Univariate and multivariate shrinkage estimators for costs and effects are proposed, then compared with one another and to the traditional methods in a simulation study. The methods are illustrated using data from a multinational trial evaluating the cost-effectiveness of three thrombolytic drug regimens in patients with acute myocardial infarction. PMID- 15803444 TI - Inferences on the common coefficient of variation. AB - The coefficient of variation is often used as a measure of precision and reproducibility of data in medical and biological science. This paper considers the problem of making inference about the common population coefficient of variation when it is a priori suspected that several independent samples are from populations with a common coefficient of variation. The procedures for confidence interval estimation and hypothesis testing are developed based on the concepts of generalized variables. The coverage properties of the proposed confidence intervals and type-I errors of the proposed tests are evaluated by simulation. The proposed methods are illustrated by a real life example. PMID- 15803443 TI - Isomeric mono- and bis[(phosphane)gold(I)] thiocyanate complexes. AB - The solid-state IR spectrum of Me(3)PAuSCN shows two signals in the range of the C-N stretching vibrations at 2075 and 2113 cm(-1). On the basis of thoroughly tested quantum chemical ab initio calculations (MP2 level of theory) these signals have been assigned to the two isomeric forms Me(3)PAuNCS and Me(3)PAuSCN. The molecular structures, the vibrational frequencies, and the relative energies of the two species have been calculated and the results compared with the experimental IR data. Treatment of Me(3)PAuSCN with equimolar quantities of [(Me(3)P)Au](+)[SbF(6)](-) in CH(2)Cl(2) at -78 degrees C gives the dinuclear reaction product [C(7)H(9)Au(2)NP(2)S](+)[SbF(6)](-) in high yields. A comparison of results of ab initio calculations and IR data suggest that at least three isomeric cationic species [(R(3)PAu)(2)NCS](+), [(R(3)PAu)(2)SCN](+) and [(R(3)PAu)SCN(AuPR(3))](+) are present, the second and third being the predominant components. The structures and vibrational frequencies of all three species have been calculated. The relative energies in the gas phase and in solution are discussed and compared with the corresponding data of the experimental IR spectra. PMID- 15803445 TI - Purification of carbon-11 PET radiotracers from unlabeled precursors by preparative HPLC and SPE. AB - A general methodology for the rapid purification of carbon-11 positron emission tomography (PET) radiotracers from radiolabeling reaction mixtures has been developed. Preparative HPLC and solid-phase extraction (SPE) techniques are described which can separate some commonly used radiopharmaceuticals such as [(11)C]raclopride, [(11)C]beta-CFT and [(11)C]choline from their unlabeled precursors. PMID- 15803446 TI - High-throughput determination of atrasentan in human plasma by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. AB - Atrasentan (A-147627) is an endothelin antagonist receptor being developed at Abbott Laboratories for the treatment of prostate cancer. A quick and sensitive method for the determination of atrasentan in human plasma has been developed and validated using high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. A dual-column, single mass spectrometer system is used to provide a reliable and routine means to increase sample throughput. The analytical method involves liquid-liquid extraction and internal standard (A-166790). The plasma samples and internal standard are acidified with 0.3 M hydrochloric acid prior to being extracted into 1:1 (v/v) hexanes--methyl t butyl ether. The organic extract was evaporated to dryness using heated nitrogen stream and reconstituted with mobile phase. Atrasentan and internal standard were separated with no interference in a Zorbax SB-C(18) analytical column with 2.1 x 50 mm, 5 microm, and a Zorbax C(8) guard column using a mobile phase consisting of 50:50 (v:v) acetonitrile--0.05 M ammonium acetate, pH 4.5, at a flow rate of 0.30 mL/min to provide 4 min chromatograms. For a 250 microL plasma sample volume, the limit of quantitation was approximately 0.3 ng/mL. The calibration was linear from 0.30 to 98.0 ng/mL (r(2) > 0.995). A significant advantage of the method is the ability to employ parallel HPLC separations with detection by a single MS/MS system to provide sensitivity and selectivity sufficient to achieve robust analytical results with a lower limit of quantitation of 0.30 ng/mL and high throughput. PMID- 15803447 TI - On the prediction and prevention of myocardial infarctions: models based on retrospective and doubly censored prospective data. AB - Early detection of coronary heart disease (CHD) in its pre-clinical stages may offer a way to reduce the impact of this endemic disease on society. Coronary calcification accompanies the development of an atherosclerotic plaque, the pathological substrate of CHD, and its identification is currently possible by means of electron beam tomography (EBT). This is a non-invasive imaging test that quantifies the extent of coronary artery plaque calcification by means of a calcium score. In this study, we show that an age-sex based calcium score percentile (CS%) provides an invaluable predictor for myocardial infarction (MI), and examine how CS% is related to traditional risk factors. We study two separate groups of patients: 172 patients who underwent EBT screening after surviving an MI (retrospective group); and 676 asymptomatic subjects who were screened and followed for several years for the occurrence of an MI (prospective group). We use CS% with traditional risk factors in logistic regression models to: (1) compare patients in the retrospective and prospective groups, and (2) develop a mortality model for MIs that occurred in the prospective group. These logistic regressions are used to develop a joint model for the relative log-odds of an MI, which is non-linear in the covariates of the mortality model. We also use baseline covariates in the prospective group to fit an event-time regression model and estimate probabilities for 2 and 3 year exposure periods. The event time regression provides independent estimates of the relative odds that are associated with risk factors. CS%, smoking, and their interaction were preeminent as predictors in the joint model for the relative odds of an MI and in the event time regression, although the effects of smoking and CS% were generally subadditive. These models provide important information on the risks associated with elevated CS% levels. PMID- 15803448 TI - Simultaneous determination of amantadine and rimantadine by HPLC in rat plasma with pre-column derivatization and fluorescence detection for pharmacokinetic studies. AB - We investigated simultaneous high-performance liquid chromatographic (HPLC) determination of amantadine hydrochloride (AMA) and rimantadine hydrochloride (RIM) levels in rat plasma after fluorescent derivatization with o-phthalaldehyde and 2-mercaptoethanol. Afterwards, the method was applied to determine their pharmacokinetics. The retention times of AMA and RIM derivatives were 12.6 and 22.2 min and the lower limits of detection were 0.025 and 0.016 microg/mL, respectively. The coefficients of variation for intra- and inter-day assay of AMA and RIM were less than 5.1 and 7.6%, respectively. After i.v. administration of AMA or RIM to rats, the total body clearance and distribution volume at the steady-state of RIM were higher than those of AMA. Bioavailability of AMA and RIM was 34.9 and 37.2%, respectively. When AMA and RIM were p.o. co-administered, the area under the plasma concentration--time curve of RIM was significantly lower than that after RIM alone. On the other hand, pharmacokinetic parameters of AMA did not significantly change. These results indicate that our HPLC assay is simple, rapid, sensitive and reproducible for simultaneously determining AMA and RIM concentrations in rat plasma and is applicable to their pharmacokinetic studies. Also, co-administration of AMA and RIM may result in the lack of pharmacological effects of RIM. PMID- 15803449 TI - Rapid and simple method for determination of Nepsilon-(carboxymethyl)lysine and Nepsilon-(carboxyethyl)lysine in urine using gas chromatography/mass spectrometry. AB - A new procedure was developed to determine in urine the concentrations of N(epsilon)-(carboxymethyl)lysine (CML) and N(epsilon)-(carboxyethyl)lysine (CEL), the major products of oxidative modification of glycated proteins, to assess levels of oxidative stress in physiological systems. The urine samples were acetonitrile-deproteinized, then derivatized by ethylchloroformate, and N(O,S) ethoxycarbonyl ethyl esters of amino acids were analysed by isotope dilution gas chromatography/mass spectrometry. Recovery averaged 89%. Linearity was excellent (r = 0.998-0.999) in the 0.5-25 micromol/L range for CML and CEL. The limit of detection of this assay was 0.1 micromol/L (corresponding to 0.20 pmol of CML or CEL on column). Intra-day and inter-day precisions were likewise excellent, with relative standard deviations <4.63 and <6.15%, respectively. Accuracy of CML and CEL determination (15 micromol/L) was 2.9 and 5.9% of the estimated theoretical value. The time from obtaining the urine sample to determination of the concentration from the chromatographic peak was 80 min or less. This method is sensitive, reproducible, accurate, relatively cheap and very simple. It can be useful for laboratories involved in the diagnosis and monitoring of age-related chronic diseases. PMID- 15803450 TI - Assessing the effect of interventions in the context of mixture distributions with detection limits. AB - Many quantitative assay measurements of metabolites of environmental toxicants in clinical investigations are subject to left censoring due to values falling below assay detection limits. Moreover, when observations occur in both unexposed individuals and exposed individuals who reflect a mixture of two distributions due to differences in exposure, metabolism, response to intervention and other factors, the measurements of these biomarkers can be bimodally distributed with an extra spike below the limit of detection. Therefore, estimating the effect of interventions on these biomarkers becomes an important and challenging problem. In this article, we present maximum likelihood methods to estimate the effect of intervention in the context of mixture distributions when a large proportion of observations are below the limit of detection. The selection of the number of components of mixture distributions was carried out using both bootstrap-based and cross-validation-based information criterion. We illustrate our methods using data from a randomized clinical trial conducted in Qidong, People's Republic of China. PMID- 15803451 TI - The effects of ligand chain length, salt concentration and temperature on the adsorption of bovine serum albumin onto polypropyleneglycol-Sepharose. AB - The interaction thermodynamics associated with bovine serum albumin adsorption on polypropylene glycol (n=3)-Sepharose CL-6B and polypropylene glycol (n=7) Sepharose CL-6B, using ammonium sulfate as the modulator was studied. Analysis of data under linear conditions was accomplished with the stoichiometric displacement retention model, preferential interaction approach and van't Hoff plots applied to HIC systems. Preferential interaction analysis indicated a strong entropic driving force under linear conditions, due to the release of a large amount of solvent on adsorption. In contrast, flow microcalorimetry under overloaded conditions showed that the adsorption of bovine serum albumin may be entropically or enthalpically driven. It is postulated that adsorption in the nonlinear region is influenced by the degree of water release, protein-protein interactions on the surface, reorientation of ligand, and conformational changes in the protein. PMID- 15803452 TI - Quantitative determination of alkylated quaternary amines and their n hydroxylated metabolites in an enzyme incubation matrix by liquid chromatography electrospray ionization mass spectrometry. AB - A simple, rapid and sensitive reversed-phase liquid chromatography method coupled to electrospray ionization mass spectrometry has been developed for studying the in vitro metabolism of the long-chain quaternary ammonium compounds dodecyltrimethylamine, tetradecyltrimethylamine and hexadecyltrimethylamine. Samples were prepared from the biological matrix by a simple protein precipitation stage. The separation was performed using a BDS Hypersil C8 3 microm particle size (100 x 3 mm i.d.) column with a fast gradient separation (60% B to 100% B) using a mobile phase of 10 mm aqueous ammonium acetate (pH 4.0, with 0.06% triethylamine; (A)-acetonitrile (B) at 0.7 mL min(-1). To minimize contamination of the MS source a switching value was used to divert the solvent front to waste. Decylammonium bromide was used as the internal standard and analytes were identified and quantified by positive ion electrospray selected ion monitoring of their intact molecular cations. The assay had a limit of quantitation of 0.25 microm (6.25 pmol on column) and was linear over the range 0.25--100 microm assay concentration for this series of long-chain quaternary amines. The precision of intra- and inter-day assays was better than 19% and the accuracy was between 93 and 109%. The method was used to assess the in vitro metabolism of the quaternary amines by wild-type cytochrome P450 enzyme CYP 4 A 1 and mutants in an artifical membrane system. PMID- 15803454 TI - Distal enhancer of the mouse FGF-4 gene and its human counterpart exhibit differential activity: critical role of a GT box. AB - Previous studies have shown that there is a strict requirement for fibroblast growth factor-4 (FGF-4) during mammalian embryogenesis, and that FGF-4 expression in embryonic stem (ES) cells and embryonal carcinoma (EC) cells are controlled by a powerful downstream distal enhancer. More recently, mouse ES cells were shown to express significantly more FGF-4 mRNA than human ES cells. In the work reported here, we demonstrate that mouse EC cells also express far more FGF-4 mRNA than human EC cells. Using a panel of FGF-4 promoter/reporter gene constructs, we demonstrate that the enhancer of the mouse FGF-4 gene is approximately tenfold more active than its human counterpart. Moreover, we demonstrate that the critical difference between the mouse and the human FGF-4 enhancer is a 4 bp difference in the sequence of an essential GT box. Importantly, we demonstrate that changing 4 bp in the human enhancer to match the sequence of the mouse GT box elevates the activity of the human FGF-4 enhancer to the same level as that of the mouse enhancer. We extended these studies by examining the roles of Sp1 and Sp3 in FGF-4 expression. Although we demonstrate that Sp3, but not Sp1, can activate the FGF-4 promoter when artificially tethered to the FGF-4 enhancer, we show that Sp3 is not essential for expression of FGF-4 mRNA in mouse ES cells. Finally, our studies with human EC cells suggest that the factor responsible for mediating the effect of the mouse GT box is unlikely to be Sp1 or Sp3, and this factor is either not expressed in human EC cells or it is not sufficiently active in these cells. PMID- 15803455 TI - Chromatography in authenticity and traceability tests of vegetable oils and dairy products: a review. AB - The new applications of various chromatographic techniques such as gas-liquid chromatography, high-performance liquid chromatography and electrophoretic methods employed for the analysis in macro- and micro-components in vegetable oils and dairy products are compiled and critically evaluated. The employment of these methods for authenticity tests and traceability is discussed. PMID- 15803456 TI - Developmental capability of denuded bovine oocyte in a co-culture system with intact cumulus-oocyte complexes: role of cumulus cells, cyclic adenosine 3',5' monophosphate, and glutathione. AB - Cumulus oophorus cells have been implicated in the regulation of female gamete development, meiotic maturation, and oocyte-sperm interaction. Nevertheless, the specific role of cumulus cells (CCs) during the final stages of oocyte maturation and fertilization processes still remains unclear. Several studies have been conducted in order to clarify the role of follicular cells using culture systems where denuded oocytes (DOs) were co-cultured with isolated CCs, or in the presence of conditioned medium. However, those attempts were ineffective and the initial oocyte competence to become a blastocyst after fertilization was only partially restored. Aim of the present study was to analyze the effect of the interactions between somatic cells and the female gamete on denuded oocyte developmental capability using a system of culture where CCs were present as dispersed CCs or as intact cumulus-oocyte complexes (COCs) in co-culture with oocytes freed of CC investment immediately after isolation from the ovary. Moreover, we analyzed the specific role of cyclic adenosine 3'-5' monophosphate (cAMP) and glutathione (GSH) during FSH-stimulated maturation of denuded oocyte co-cultured with intact COCs. Our data confirm that denuded oocyte has a scarce developmental capability, and the presence of dispersed CCs during in vitro maturation (IVM) does not improve their developmental competence. On the contrary, the co-presence of intact COCs during denuded oocyte IVM partially restores their developmental capability. The absence of CCs investment causes a drop of cAMP content in DOs at the beginning of IVM and the addition of a cAMP analog in the culture medium does not restore the initial oocyte developmental competence. The relative GSH content of denuded oocyte matured in presence of intact COCs is consistent with the partial recovery of their developmental capability. However, the complete restoration of a full embryonic developmental potential is achieved only when DOs are co-cultured with intact COCs during both IVM and in vitro fertilization (IVF). Our results suggest that the direct interaction between oocyte and CCs is not essential during IVM and IVF of denuded oocyte. We hypothesize that putative diffusible factor(s), produced by CCs and/or by the crosstalk between oocyte and CCs in the intact complex, could play a key role in the acquisition of developmental competence of the denuded female gamete. PMID- 15803457 TI - Intraoviductal introduction of plasmid DNA and subsequent electroporation for efficient in vivo gene transfer to murine oviductal epithelium. AB - Various growth factors and proteins produced by oviductal cells have been demonstrated to interact with developing embryos. However, little is known concerning the function of mammalian oviducts at the molecular biological level. This may be partly due to lack of efficient gene transfer to oviductal cells. In this study, we developed an efficient method for transfection of oviductal epithelium using in vivo electroporation (EP) in mice. One microliter of solution containing enhanced green fluorescent protein (EGFP) expression plasmid (0.5 microg) and 0.05% trypan blue (TB) were directly introduced into the ampulla of the eCG-hCG-treated B6C3F1 females at embryonic day (E) 0.6 of pregnancy (corresponding to 14:00-15:00 of the day the plug was recognized). The entire oviduct was then electroporated using tweezer-type electrodes attached to a T820 electroporator (BTX Genetronics, Inc., San Diego, CA) with eight square-wave pulses, 50 V in strength and 50 msec in duration. On E 3.4, embryos at morula/early blastocyst stages were collected and their number, morphology, and EGFP-derived fluorescence recorded. Fluorescence in oviducts was also examined. In some cases, these fluorescent oviducts were subjected to cryostat sectioning. Strong fluorescence was observed in some of the oviductal epithelia, with a maximum level of 36%. Neither the number nor morphology of the collected embryos was affected by EP. Some embryos possessed fluorescence in the blastocoel, but not cytoplasm, suggesting incorporation of EGFP present in the oviductal luminal fluid. This system may enable development of new factors regulating development of preimplantation embryos and offers the prospect of a new approach to understanding oviductal function. PMID- 15803458 TI - Specific maternal transcripts in bovine oocytes and cleavaged embryos: identification with novel DDRT-PCR methods. AB - We used annealing control primer (ACP)-based differential display reverse transcription polymerase chain reaction (DDRT-PCR) to isolate differentially expressed amplicons in bovine germinal vesicle (GV) stage oocytes, 8-cell stage embryos produced in vitro, and blastocyst stage embryos produced in vitro. Four expressed sequence tags (ESTs) of genes that were specifically and predominantly expressed in GV oocytes were cloned and sequenced. We have used a fluorescence monitored real-time quantitative PCR (qPCR) to quantify and analyzed the temporal expression of the target differentially expressed transcripts throughout the preimplantation stages from oocytes to blastocysts. The cloned genes or ESTs all exhibited significant sequence similarity with known bovine genes (98%-100%; DNCL1 and ZP2) or ESTs (81%-97%; FANK1 and GTL3) of other species. As revealed by real-time qRT-PCR, DNCL1, FANK1, GTL3, and ZP2 transcripts were observed in the GV stage oocytes and expression gradually decreased up to the 8-cell stage embryo and the transcripts were not detected in later stages. Similarly, upregulation was observed in GV stage mouse oocytes and metaphase II, suggesting that these four differentially expressed orthologous genes play important roles in early preimplantation, as maternally-derived transcripts. PMID- 15803459 TI - Actions of progesterone on uterine immunosuppression and endometrial gland development in the uterine gland knockout (UGKO) ewe. AB - In ewes, the uterine gland knockout (UGKO) phenotype is caused by neonatal exposure to norgestomet to arrest uterine gland development and produce an adult which has a uterus characterized by the lack of endometrial glands. Since endometrial glands in the sheep produce the lymphocyte-inhibitory protein, ovine uterine serpin (OvUS), an experiment was conducted with ewes of the UGKO phenotype to evaluate whether the inhibitory actions of progesterone on tissue rejection responses in utero are dependent upon the presence of endometrial glands. Control and UGKO ewes were ovariectomized and subsequently treated with either 100 mg/day progesterone or corn oil vehicle for 30 days. An autograft and allograft of skin were then placed in each uterine lumen and treatments were continued for an additional 30 days before grafts were examined for survival. All autografts survived and had a healthy appearance after histological analysis. Allografts were generally rejected in ewes treated with vehicle but were present for hormone-treated ewes, regardless of uterine phenotype. Analysis of the histoarchitecture and protein synthetic capacity of the uterus revealed that progesterone induced differentiation of endometrial glands and synthesis and secretion of OvUS in UGKO ewes. The UGKO ewes had reduced density of CD45R+ lymphocytes in the endometrial epithelium and there was a tendency for progesterone to reduce this effect in luminal epithelium. Taken together, results confirm the actions of progesterone to inhibit graft rejection response in utero. Responses of UGKO ewes to progesterone indicate that the hormone can induce de novo development and differentiation of endometrial glands, at least when skin grafts are in the uterus. PMID- 15803460 TI - PLAC1 expression increases during trophoblast differentiation: evidence for regulatory interactions with the fibroblast growth factor-7 (FGF-7) axis. AB - PLAC1 is a recently described, trophoblast-specific gene that localizes to a region of the X-chromosome important in placental development. Immunohistochemical analysis demonstrated that PLAC1 polypeptide localizes to the differentiated syncytiotrophoblast throughout gestation (8-41 weeks) as well as a small population of villous cytotrophoblasts. Consistent with these observations, quantitative RT-PCR demonstrated that PLAC1 mRNA increases more than 300-fold during cytotrophoblast differentiation in culture to form syncytiotrophoblasts. Agents known to be relevant to trophoblast differentiation were then tested for the ability to influence PLAC1 expression. Fibroblast growth factor-7 (FGF-7), also known as keratinocyte growth factor (KGF), stimulated PLAC1 mRNA expression approximately two-fold in the BeWo(b30) trophoblast cell line. FGF-7 stimulation was significantly inhibited by PD-98059 and wortmannin suggesting mediation via MAP kinase and PI-3 kinase-dependent signaling pathways. Interestingly, epidermal growth factor (EGF) treatment of trophoblasts had no effect on PLAC1 expression alone, but potentiated the effect of FGF-7, suggesting the presence of a regulatory interaction of the two growth factors. FGF-7 and its receptor, FGFR 2b, exhibited spatial overlap with PLAC1 suggesting these regulatory interactions are physiologically relevant during gestation. These data demonstrate PLAC1 expression is upregulated during trophoblast differentiation, localizing primarily to the differentiated syncytiotrophoblast. Furthermore PLAC1 expression is specifically regulated by peptide growth factors relevant to trophoblast differentiation. PMID- 15803461 TI - The role of anion channels and Ca2+ in addition to K+ channels in the physiological volume regulation of murine spermatozoa. AB - Studies in the human, transgenic mice, and cattle indicate that sperm cell volume regulation plays an important role in male fertility as spermatozoa encounter a hypo-osmotic challenge upon ejaculation into the female tract. Physiological regulatory volume decrease (RVD) was examined using flow cytometry in murine sperm released into incubation medium mimicking uterine osmolality and including putative channel inhibitors. The involvement of K+ channels was indicated by the recovery of volume regulation by the K+ ionophore valinomycin in defective sperm from infertile transgenic mice, and from blockage of RVD by quinine in normal sperm. However, in neither case was the recovery complete. The involvement of volume-sensitive osmolyte and anion channels (VSOAC) were investigated using blockers effective in other cell types. NPPB (5-nitro-2(3-phenylpropylamino) benzoic acid) and tamoxifen inhibited RVD but SITS (4-acetamido-4'-isothiocyanato stilbene-2,2'-disulphonic acid) at 0.4 and 1 mM had no effect whereas DIDS (di isothiocyanato-stilbene-2,2'-disulphonic acid) at 1 mM enhanced RVD. Verapamil, but not another P-glycoprotein antagonist cyclosporin, caused sperm swelling which persisted in the presence of valinomycin, in Ca2+-free medium and in the presence of thapsigargin, but swelling was abolished by the Ca2+ ionophore A23187. Nifedipine was slightly effective in blocking RVD. Analysis by Western blotting failed to reveal ClC-2 and ClC-3 members of the chloride channel family in murine or rat sperm proteins despite signal bands in positive tissue controls. These findings implicate the involvement of some unidentified VSOAC in sperm volume regulation, which is probably Ca+-dependent. PMID- 15803462 TI - Phosphorylcholine is favorable for antibody production from hybridoma cells. AB - Growth of antibody-secreting hybridomas requires special conditions such as serum free defined media containing growth factors and vitamins. However, the surface on which these cells can proliferate has been shown to play an important role. Phosphorylcholine (PC)-based polymers are zwitterionic compounds with nonbiofouling properties. These polymers are characterized by having reduced protein absorption properties. Our aim was to determine whether well-established hybridoma cell lines were able to proliferate and produce measurable amounts of monoclonal antibodies when grown on PC-polymer-coated surfaces. Comparative experiments using four well-known hybridoma cell lines (PAb421, PAb246, PAb1801 which recognize p53, and PAb280 which recognizes SV40 small t antigen) grown on PC-polymer-coated, uncoated, and two commercially available tissue culture plates showed that PC-polymer-coated plates were more efficient than uncoated plates in sustaining cell growth and monoclonal antibody production/secretion as defined by growth assays and ELISA. Also, results demonstrated that PC-polymer-coated plates were able to perform better than commercially available plates. These observations suggest that PC polymers could be used as an alternative, efficient surface coating to grow hybridoma cell lines and allow detectable antibody secretion. PMID- 15803463 TI - Template-directed self-assembly and growth of insulin amyloid fibrils. AB - The formation of amyloid aggregates in tissue is a pathological feature of many neurodegenerative diseases and type II diabetes. Amyloid deposition, the process of amyloid growth by the association of individual soluble amyloid molecules with a pre-existing amyloid template (i.e., plaque), is known to be critical for amyloid formation in vivo. The requirement for a natural amyloid template, however, has made amyloid deposition study difficult and cumbersome. In the present work, we developed a novel, synthetic amyloid template by attaching amyloid seeds covalently onto an N-hydroxysuccinimide-activated surface, where insulin was chosen as a model amyloidogenic protein. According to ex situ atomic force microscopy observations, insulin monomers in solution were deposited onto the synthetic amyloid template to form fibrils, like hair growth. The fibril formation on the template occurred without lag time, and its rate was highly accelerated than in the solution. The fibrils were long, over 2 mum, and much thinner than those in the solution, which was caused by limited nucleation sites on the template surface and lack of lateral twisting between fibrils. According to our investigations using thioflavin T-induced fluorescence, birefringent Congo red binding, and circular dichroism, fibrils grown on the template were identified to be amyloids that formed through a conformational rearrangement of insulin monomers upon interaction with the template. The amyloid deposition rate followed saturation kinetics with respect to insulin concentration in the solution. The characteristics of amyloid deposition on the synthetic template were in agreement with previous studies performed with human amyloid plaques. It is demonstrated that the synthetic amyloid template can be used for the screening of inhibitors on amyloid deposition in vitro. PMID- 15803464 TI - Immobilized N-alkylated polyethylenimine avidly kills bacteria by rupturing cell membranes with no resistance developed. AB - Several critical mechanistic and phenomenological aspects of the microbicidal surface coatings based on immobilized hydrophobic polycations, previously developed by us, are addressed. Using Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacteria, remarkable bactericidal action (up to a 10(9)-fold reduction in live bacteria count in the surface-exposed solution and a 100% inactivation of the surface-adhered bacteria) of an amino glass slide covalently derivatized with N-hexyl,methyl-polyethylenimine (PEI) is found to be due to rupturing bacterial cell membranes by the polymeric chains. The bacteria fail to develop noticeable resistance to this lethal action over the course of many successive generations. Finally, the immobilized N-alkyl-PEI, while deadly to bacteria, is determined to be harmless to mammalian (monkey kidney) cells. PMID- 15803465 TI - Growth characteristics of Arthrospira platensis cultured inside a new close-coil photobioreactor incorporating a mandrel to control culture temperature. AB - The aim of this study was to investigate Arthrospira growth inside a new CCP incorporating a mandrel for culture temperature control. Some hydrodynamic aspects and photobioreactor performances were investigated as well. The bioreactor incorporated A. platensis grown under batch and semicontinuous conditions. Two systems were used to recycle Arthrospira cultures: a peristaltic pump and an airlift system. When the pump recycled the culture, we achieved a very high Dean number (De=3,950), which decreased a great deal when the pump was replaced with the airlift system. During outdoor Arthrospira batch growth, a cell concentration of 16.4 g (DW)l-1 was reached after 9 days. However, the maximum chlorophyll content of the biomass (2.0% of DW) was achieved on the fifth and sixth days. The highest daily biomass output rate was obtained using the airlift system, when the CCP was operated under a semicontinuous regime: the gross output rate was 2.85+/-0.37 g (DW) l-1 d-1 and the net was 2.32+/-0.11 g (DW) l-1 d-1. The advantages of the airlift system may be due to the low concentration of oxygen built up inside Arthrospira culture and the lack of cell damage due to the pump system. Thus, oxygen and pump stress may have been avoided. PMID- 15803466 TI - Localization of receptors in lipid rafts can inhibit signal transduction. AB - Processes of cell survival, division, differentiation, and death are guided by the binding of signal molecules to receptors, which activates intracellular signaling networks and ultimately elicits genetic, biochemical, or biomechanical responses within the cell. While intracellular mechanisms for these processes have been well studied, little attention has been given to the role extracellular ligand transport and binding may play in signal initiation. Recent studies have found that the localization of receptors in lipid rafts is critical for the functions of many signaling pathways. By concentrating membrane components, rafts may promote essential interactions for signaling. Lipid rafts can also have negative effects on signaling, but mechanisms remain elusive. We propose that raft-mediated receptor clustering can reduce signaling by prolonging the diffusion of ligands to their receptors. We quantify this effect using a simple diffusion-limited binding model that accounts for the spatial distribution of lipid rafts and receptors on the cell surface. We find that receptor clustering can reduce the apparent rate of receptor binding by up to 80%, consistent with observed increases in epidermal growth factor (EGF) binding by up to 100% following disruption of lipid rafts (Pike and Casey 2002 Biochemistry 41:10315 10322; Roepstorff et al. 2002 J Biol Chem 277:18954-18960). Failure to account for the effects of receptor clustering on rates of ligand binding can skew the interpretation of current methods of cancer diagnosis and treatment. Finally, we discuss how the activation of particular signaling pathways can change over time, depending, in part, on the overall level and spatial distribution of the receptors. PMID- 15803467 TI - Fed-batch culture of a metabolically engineered Escherichia coli strain designed for high-level succinate production and yield under aerobic conditions. AB - An aerobic succinate production system developed by Lin et al. (Metab Eng, in press) is capable of achieving the maximum theoretical succinate yield of 1.0 mol/mol glucose for aerobic conditions. It also exhibits high succinate productivity. This succinate production system is a mutant E. coli strain with five pathways inactivated: DeltasdhAB, Delta(ackA-pta), DeltapoxB, DeltaiclR, and DeltaptsG. The mutant strain also overexpresses Sorghum vulgare pepc. This mutant strain is designated HL27659k(pKK313). Fed-batch reactor experiments were performed for the strain HL27659k(pKK313) under aerobic conditions to determine and demonstrate its capacity for high-level succinate production. Results showed that it could produce 58.3 g/l of succinate in 59 h under complete aerobic conditions. Throughout the entire fermentation the average succinate yield was 0.94+/-0.07 mol/mol glucose, the average productivity was 1.08+/-0.06 g/l-h, and the average specific productivity was 89.77+/-3.40 mg/g-h. Strain HL27659k (pKK313) is, thus, capable of large-scale succinate production under aerobic conditions. The results also showed that the aerobic succinate production system using the designed strain HL27659k(pKK313) is more practical than conventional anaerobic succinate production systems. It has remarkable potential for industrial-scale succinate production and process optimization. PMID- 15803468 TI - Structured model for denitrifier diauxic growth. AB - We present a model for diauxic growth of denitrifying bacteria in which nitrate reductase synthesis kinetics dominate the overall growth kinetics. The model is based on the assumption of the existence of a nitrate respiration operon, thereby linking the rate of nitrate uptake to the activity of nitrate reductase. We show that this approach can model diauxic growth of Pseudomonas denitrificans by conducting experiments in which nitrate reductase activity was measured during both lag and ensuing exponential growth phases. We consistently observed the pattern of low nitrate reductase enzyme activity during the lag phase, increasing before the onset of growth. By fitting model parameters we were able to successfully match experimental data for growth, nitrate uptake, and enzyme activity level. PMID- 15803469 TI - Stable production of hyaluronic acid in Streptococcus zooepidemicus chemostats operated at high dilution rate. AB - Hyaluronic acid is routinely produced through fermentation of both Group A and C streptococci. Despite significant production costs associated with short fermentations and removal of contaminating proteins released during entry into stationary phase, hyaluronic acid is typically produced in batch rather than continuous culture. The main reason is that hyaluronic acid synthesis has been found to be unstable in continuous culture except at very low dilution rates. Here, we investigated the mechanisms underlying this instability and developed a stable, high dilution rate (0.4 h-1) chemostat process for both chemically defined and complex media operating for more than 150 h of production. In chemically defined medium, the product yield was 25% higher in chemostat cultures than in conventional batch culture when arginine or glucose was the limiting substrate. In contrast, glutamine limitation resulted in higher ATP requirements and a yield similar to that observed in batch culture. In complex, glucose limited medium, ATP requirements were greatly reduced but biomass synthesis was favored over hyaluronic acid and no improvement in hyaluronic acid yield was observed. The successful establishment of continuous culture at high dilution rate enables both commercial production at reduced cost and a more rational characterization and optimization of hyaluronic acid production in streptococci. PMID- 15803470 TI - High-yield retroviral production using a temperature-modulated two-stage operation. AB - For clinical trials, large amounts of high-titer retroviral supernatants are required. However, retroviral concentration is relatively low compared with other viral vectors. Moreover, less than half of retroviral vectors suspended in a collected supernatant are infectious because of their short half-lives. In this study, a culture medium of ecotropic retrovirus-producing GP + E86/LNCX cells in tissue culture dishes was circulated through a reservoir, which was arranged with an incubator or ice-bath stage. Titers determined from the retroviral supernatant circulated through an ice-cold reservoir increased for a week from the beginning of retroviral production, while the titers from static production with circulation through the 37 degrees C reservoir reached a plateau after 3 days of retroviral production. After 5 days, 10 times more infectious retroviruses were obtained by circulating and keeping the majority of supernatant longer in the cold reservoir than in the production vessel at 37 degrees C in comparison with the number collected from the static tissue culture dish without circulating the culture medium. Furthermore, the concentration of transduction inhibitors in the supernatant was decreased along with the retardation of retroviral decay at low temperature. The two-stage operation developed in this study should be easily applied to large-scale bioreactors for mass production of high-titer retroviral supernatants. PMID- 15803471 TI - Transient gene expression in HEK293 cells: peptone addition posttransfection improves recombinant protein synthesis. AB - Gene expression by large-scale transfection of mammalian cells is becoming an established technology for the fast production of milligram and even gram amounts of recombinant proteins (r-proteins). However, efforts are still needed to optimize production parameters in order to maximize volumetric productivities while maintaining product quality. In this study, transfection efficiency and volumetric productivity following transient gene expression in HEK293 cells were evaluated using green fluorescent protein (GFP) and human placental secreted alkaline phosphatase (SEAP) as reporter genes. We show that a single pulse of peptones (protein hydrolysates) to the cultures performed in a low serum (1%, v/v) and in serum-free medium results in a significant increase in volumetric protein productivity. Sixteen peptones from different sources were tested and almost all of them showed a positive effect on r-protein production. This effect, however, is time- and concentration-dependent. By using Tryptone N1 (a casein peptone, TN1) to feed the cultures at 24 h posttransfection (hpt), a 2-fold increase in volumetric SEAP productivity was obtained 5 days posttransfection. This effect was shown to be equal to that obtained when the culture was fed with a supplementary 4% (v/v) of serum. The positive effect of TN1 on protein production was also demonstrated with Tie2 protein ectodomain produced in serum free medium. HPLC analysis of amino acids consumption/production during control batch and TN1 pulse culture showed some major differences in amino acid metabolism when using TN1 pulse. Asparagine, glycine, histidine, threonine, leucine, and valine show accumulation in the medium over the cultivation period instead of being consumed as observed in unfed sample (except for asparagine, which remained unchanged). Isoleucine, tyrosine, methionine, and phenylalanine all remained unchanged or slightly fluctuated in TN1-fed culture after the feeding pulse, while they were all steadily consumed in the control run. The relative abundance of SEAP's mRNA suggests that the improvement in protein yield results both from an increase of the translational activity and transcription efficiency. Further understanding of mechanisms by which amino acids/peptides regulate transcriptional and translational machinery in mammalian cells should facilitate the design of new strategies for the improvement of r-protein production by large-scale transfection. PMID- 15803472 TI - Cranial base angulation and growth of the human fetal pharynx. AB - The upper airway controls the mechanisms of breathing, swallowing, and vocalization. Information on its normal ontogenetic associations is therefore crucial for understanding human speech evolution as well as the developmental etiology of speech abnormalities. Of particular interest to evolutionary morphologists is the proposed structural association between the upper airway and the cranial base. The present study aims to elucidate whether cranial base angulation is linked to growth of the upper airway during human fetal life. Forty postmortem human fetuses ranging from 10 to 29 weeks of gestation were imaged with high-resolution magnetic resonance imaging. Measurements of basicranial angle, hyoid depth, hormion height, and pharyngeal area were taken from sagittal images. Results show a significant correlation between the internal and external basicranial angles but the correlation between relative hyoid depth and internal cranial base angle was not significant. Findings show a significant correlation between increases in the size of the oropharynx relative to the nasopharynx and increases in the internal cranial base angle (retroflexion). These findings give limited support (rrank=0.46; P<0.01) for the hypothesis that restructuring of the upper airway underlies basicranial retroflexion during prenatal life. PMID- 15803473 TI - Fractionation of bovine serum albumin and monoclonal antibody alemtuzumab using carrier phase ultrafiltration. AB - Protein transmission and hence selectivity of separation can be significantly affected by solution pH and ionic strength in protein fractionation using ultrafiltration. Using parameter scanning ultrafiltration, the transmission of bovine serum albumin (BSA) and monoclonal antibody alemtuzumab (Campath-1H) through 300 kDa polyethersulfone (PES) ultrafiltration membranes were studied over a range of pH and salt concentrations, with focus on the likely conditions for achieving "reverse selectivity," i.e., obtaining purified alemtuzumab (approximately 155 kDa) in the permeate. Experimental results demonstrate that the reverse selectivity could be obtained by manipulating the operating conditions such as the solution pH, ionic strength, permeate flux, and system hydrodynamics. With a two-stage batch ultrafiltration process under suitable conditions, the monoclonal antibody alemtuzumab with a purity of > 98% was obtained in the permeate from a feed solution initially containing 0.50 g/l each of BSA and alemtuzumab. Further purity can be expected by selecting more suitable membranes and optimizing operating conditions. PMID- 15803474 TI - Vesicular glutamate transporter-1 colocalizes with endogenous opioid peptides in axon terminals of the rat locus coeruleus. AB - We have previously shown that a subset of axon terminals in the locus coeruleus (LC) containing methionine(5)-enkephalin (ENK) forms type I (asymmetric-type) synaptic specializations that are characteristic of excitatory-type transmitters. In addition, we previously provided ultrastructural evidence showing that ENK is colocalized with glutamate using a combination of pre- and postembedding immunohistochemistry. To examine cellular substrates for interactions between glutamate and other endogenous opioid peptides in the LC, we examined the localization of the vesicular glutamate transporter 1 (VGLUT1), a transporter protein involved in the accumulation of the transmitter glutamate into synaptic vesicles, with either ENK or preprodynorphin (ppDYN). Dual-immunofluorescence and electron microscopy showed prominent coexistence of VGLUT1 and ENK in varicose processes of the LC, confirming our previous report using postembedding immunolabeling for glutamate. Likewise, VGLUT1 and ppDYN were identified in common varicose processes in the LC using confocal fluorescence microscopy. Immunoelectron microscopy using gold-silver labeling for VGLUT1 and peroxidase labeling for ppDYN established that this endogenous opioid peptide also colocalizes with glutamate transporters. The majority of these formed asymmetric type synapses. Taken together, these results demonstrate that excitatory LC afferents are enriched with endogenous opioid peptides and are positioned to modulate LC neuronal activity dually. PMID- 15803475 TI - Embryonic cerebrospinal fluid regulates neuroepithelial survival, proliferation, and neurogenesis in chick embryos. AB - Early in development, the behavior of neuroepithelial cells is controlled by several factors, which act in a developmentally regulated manner. Diffusible factors are secreted locally by the neuroepithelium itself, although other nearby structures may also be involved. Evidence suggests a physiological role for the cerebrospinal fluid in the development of the brain. Here, using organotypic cultures of chick embryo neuroepithelial explants from the mesencephalon, we show that the neuroepithelium in vitro is not able to self-induce cell survival, replication, and neurogenesis. We also show that the embryonic cerebrospinal fluid (E-CSF) promotes neuroepithelial stem cell survival and induces proliferation and neurogenesis in mesencephalic explants. These data strongly suggest that E-CSF is involved in the regulation of neuroepithelial cells behavior, supporting the hypothesis that this fluid plays a key role during the early development of the central nervous system. PMID- 15803476 TI - Ventral abdominal wall dysmorphogenesis of Msx1/Msx2 double-mutant mice. AB - Msx1 and Msx2 genes encode the homeodomain transcription factors. Several gene knockout mice and expression studies suggest that they possess functionally redundant roles in embryogenesis. In this study, we revealed that Msx1 and Msx2 were expressed during ventral body wall formation in an overlapping manner. Msx1/Msx2 double-mutant mice displayed embryonic abdominal wall defects with disorganized muscle layers and connective tissues. These findings indicate that Msx1 and Msx2 play roles in concert during embryonic ventral abdominal wall formation. PMID- 15803477 TI - Arterial embolization using poly-N-acetyl glucosamine gel in a rat kidney model. AB - Aqueous solutions of poly-N-acetyl glucosamine (p-GlcNAc) exhibit a liquid-gel transition at physiological pH and temperature. This feature inspired the authors to conduct a study to evaluate the macro- and histological changes of rat kidneys after embolization using either p-GlcNAc gel injection into the renal artery or ligation of the renal artery. The procedures were performed in 46 rats through open abdominal surgeries. Animals were sacrificed at 3 days and at 1, 3, 5, and 8 weeks postoperatively. The results of both macro-observation and histological study showed that p-GlcNAc gels were effective in causing necrosis and subsequent fibrosis in all embolized kidneys. The data indicate that p-GlcNAc gel may have promise as an effective agent for therapeutic embolization. PMID- 15803478 TI - Venous sphincters in the choroidea of spontaneously hypertensive rats. AB - Ocular vasculopathy resulting from severe systemic hypertension affects retina, choroidea, and the optic nerve. While the pathologic changes of the arterial system, including luminal narrowing, are well documented, little is known about the ocular venous vessels in hypertension. Adult 18-week-old spontaneously hypertensive rats (SHRs) were untreated (control) or treated with lisinopril for 4 weeks; normotensive Wistar-Kyoto (WKY) rats were additional controls. The mean systemic blood pressure (MSP) was monitored chronically using telemetry. The ocular microvasculature was examined using scanning electron microscopy (SEM) of corrosion casts, histology of serial sections, and computer-based 3D reconstruction. The MSP in control SHRs (145 +/- 11.9 mmHg) was decreased to 68.1 +/- 4.9 mmHg (P < 0.001) following treatment, which was even below the baseline level of WKY (96.7 +/- 5.8) rats (P < 0.05). In addition to media thickening in arteries, the venous plexus of the choroidea in control SHRs revealed multiple tufts of smooth muscle cells (sphincters) that narrowed the lumen. Correlating to histology, SEM of casts and 3D reconstruction showed numerous constrictions and muscular tufts in veins of the choroidea, narrowing the vascular lumen up to 47%. Following antihypertensive treatment, the percentage of sphincter constriction was decreased to 6% (P < 0.001). The depth of venous sphincter contraction correlated significantly with MSP (r = 0.87). To our knowledge, these results indicate for the first time that venous sphincters occur in the choroidea of the SHRs, and that their muscularity correlates with MSP. Venous sphincters might be involved in the pathophysiology of hypertension-related ocular changes. PMID- 15803479 TI - Immunolocalization of chick periostin protein in the developing heart. AB - The process that cardiac cushions undergo to form the mature septa and valves of the adult heart is poorly understood. Periostin is an extracellular molecule that is expressed during cushion mesenchyme formation and throughout valvulogenesis. Once thought to be an osteoblast-specific factor, studies have shown this molecule is antiosteogenic. We have produced an antibody to chicken periostin and examined periostin's localization in the developing avian heart. This antibody recognized proteins from chick heart lysates around 90 kD molecular weight as predicted from the chick periostin mRNA and other periostin orthologs. Periostin immunolocalization was first evident as fibrous strands in the cushion mesenchyme. At HH25, periostin was detected on the basal surface of the trabecular endothelium and also on the endocardial epithelium of the atrioventricular cushion. We hypothesize that periostin may function in the organization of extracellular matrix molecules, providing cues necessary for attachment and spreading during the epithelial-to-mesenchymal transitions of the endocardial epithelium. Enhanced secretion of periostin in the region of delamination may directly or indirectly promote change in the myocardium that precedes or mediates delamination of the leaflet. At later stages of development (HH34-38), periostin was seen predominantly in the fibrous regions of the heart, such as the left atrioventricular valve (LAV), annulus, cardiac skeleton, and adventitia. We propose that periostin is induced by sheer stress and may be an essential molecular component for structures of the heart that undergo mechanical stress or tension during the cardiac cycle. PMID- 15803480 TI - Muscle carnitine in hypo- and hyperthyroidism. AB - Weakness is common in both hyper- and hypothyroidism, and skeletal muscle L carnitine may play a role in this regard, as suggested by studies indicating abnormal levels of carnitine in serum and urine of patients with thyroid dysfunction. Skeletal muscle samples were obtained for carnitine analysis from control subjects, and from hyperthyroid and hypothyroid patients before and after treatment. There was a significant reduction in carnitine, especially the esterified portion, in hyperthyroid individuals, with a return to normal as euthyroid status was regained. In hypothyroid patients, there was a trend for carnitine to be lower than normal and for improvement once euthyroid status was attained. Our data indicate that muscle carnitine levels are affected by both hypo- and hyperthyroidism. A decrease in muscle carnitine in both conditions may contribute to thyroid myopathy. PMID- 15803481 TI - Characteristic distribution of immunoreaction for estrogen receptor alpha in rat ameloblasts. AB - Estrogen has a diverse function, including cell proliferation and differentiation via estrogen receptors (ER), which have been reported to be the case in various tissues in addition to female reproductive organs. A recent immunocytochemical study has reported the expression of ERalpha, a subtype of ER, in rat odontoblasts, suggesting an involvement of estrogen in the differentiation of tooth-forming cells. However, there is no information on the ERalpha immunoexpression in ameloblasts. The present study was therefore undertaken to examine the localization of ERalpha immunoreaction in rat ameloblasts during amelogenesis. A computer-assisted quantitative analysis under a confocal laser scanning microscope was employed to demonstrate the stage-specific localization pattern of ERalpha immunoreaction. Immunohistochemistry of the rat enamel organ revealed ERalpha expression as nuclear localization in ameloblasts, stratum intermedium, stellate reticulum, and papillary layer, in addition to mature and immature odontoblasts. The ratio of immunopositive nuclei to total nuclei (immunopositive ratio) in ameloblasts was high at the apical loop region and gradually declined at the presecretory stage to zero at the secretory stage with statistically significant difference. The ERalpha immunolabeling pattern exhibited a periodic change at the maturation stage proper with constant higher labeling in ruffle-ended ameloblasts than in smooth-ended ameloblasts. The positive ratio was then followed by a statistically significant increase in immunolabeling thereafter. This stage-specific immunolabeling pattern during amelogenesis suggests a possible role of ERalpha in ameloblast proliferation and differentiation. PMID- 15803482 TI - Isolated vitamin E deficiency with demyelinating neuropathy. AB - A 22-year-old man, with a past history of generalized tonic-clonic seizures treated with phenobarbital, presented with spinocerebellar ataxia. The electrophysiological studies revealed a demyelinating motor-sensory neuropathy. The serum vitamin E level was low. Sural nerve biopsy revealed loss of large myelinated fibers with evidence of remyelination. Vitamin E supplementation led to clinical and electrophysiological recovery of sensory conduction and evoked potentials. Motor nerve conduction, however, showed only partial recovery. Vitamin E deficiency leading to a demyelinating neuropathy, as in the present case, suggests that the full spectrum of the disease entity is not fully defined. PMID- 15803483 TI - Phospholipase A2-dependent effects of the venom from the New Guinean small-eyed snake Micropechis ikaheka. AB - The New Guinean small-eyed snake (Micropechis ikaheka) is a cause of life threatening envenoming. Previous studies on M. ikaheka venom have indicated the presence of neurotoxins as well as myotoxins. This study examined the in vitro myotoxic effects of M. ikaheka venom and the efficacy of a polyvalent antivenom in neutralizing these effects. Venom (50 microg/ml) produced a slowly developing contracture and inhibition of direct twitches of the chick biventer cervicis nerve-muscle preparation in the presence of tubocurarine (10 microM). Myotoxicity was confirmed by subsequent histological examination of tissues. This myotoxicity was prevented by the prior addition of polyvalent snake antivenom (30 U/ml). However, the addition of antivenom (30 U/ml) 1 h after venom administration failed to reverse or prevent the further inhibition of direct twitches. In addition, venom (1-10 microg/ml) produced concentration-dependent contractions of the guinea-pig isolated ileum. These effects were dependent on phospholipase A2 (PLA2) activity of the venom as evidenced by the ability of the PLA2 inhibitor 4 bromophenacyl bromide (4-BPB; 1.8 mM) to prevent this activity. This study indicates that M. ikaheka venom causes significant myotoxicity and that polyvalent snake antivenom may be a potential treatment for the myotoxic effects in patients envenomed by this species. PMID- 15803484 TI - Placentation in the Mexican lizard Sceloporus mucronatus (Squamata: Phrynosomatidae). AB - We used light microscopy to study placental structure of the lizard Sceloporus mucronatus throughout 6 months of embryonic development. Three stages of placental development could be assigned to embryos based on the arrangement of the extraembryonic membranes. A highly vascular choriovitelline placenta was present in the embryonic hemisphere and a nonvascular bilaminar omphalopleure covered most of the abembryonic hemisphere of the egg during embryonic Stages 10 28. A chorioallantoic placenta replaced the choriovitelline placenta by embryonic Stage 29 and an omphaloplacenta covered the abembryonic hemisphere at this stage. The combination of these two placental types occurred in Stage 29-36 embryos. The final stage of placentation, embryonic Stages 37-40, was characterized by an omphalallantoic placenta in the abembryonic hemisphere and a chorioallantoic placenta in the embryonic hemisphere of the egg. The choriovitelline and chorioallantoic placentae are well vascularized, with closely apposed maternal and embryonic blood vessels. These structures are the most likely sites of respiratory exchange. In contrast, the omphaloplacenta and omphalallantoic placentae contain cuboidal or columnar epithelia and these structures may function in histotrophic exchange. Placentation of S. mucronatus is similar to that of predominantly lecithotrophic species in other squamate lineages suggesting that the evolution of this placental morphology is a response to similar factors and is independent of phylogeny. PMID- 15803485 TI - Acute exertional compartment syndrome in the setting of anabolic steroids: an unusual cause of bilateral footdrop. AB - Acute exertional compartment syndrome is the result of muscle ischemia within a tight fascial compartment. We report a 22-year-old boxer, with recent intake of anabolic steroids, who developed acute exertional compartment syndrome of the lower legs following an assault from which he had to run away. He presented with bilateral footdrop. Nerve conduction studies (NCS) and electromyography (EMG) were consistent with bilateral deep and superficial peroneal neuropathies, but magnetic resonance imaging (MRI) demonstrated hemorrhagic necrosis of the pretibial muscles. This case illustrates that the differential diagnosis for footdrop includes not only central and peripheral nervous system and muscle causes, but also compartment syndromes. PMID- 15803486 TI - Functional morphology of the mantle of Nautilus pompilius (Mollusca, Cephalopoda). AB - This study presents histological and cytological findings on the structural differentiation of the mantle of Nautilus pompilius in order to characterize the cells that are responsible for shell formation. The lateral and front mantle edges split distally into three folds: an outer, middle, and inner fold. Within the upper part of the mantle the mantle edge is divided into two folds only; the inner fold disappears where the hood is attached to the mantle. At the base of the outer fold of the lateral and front mantle edge an endo-epithelial gland, the mantle edge gland, is localized. The gland cells are distinguished by a distinct rough endoplasmic reticulum and by numerous secretory vesicles. Furthermore, they show a strong accumulation of calcium compounds, indicating that the formation of the shell takes place in this region of the mantle. Numerous synaptic contacts between the gland cells and the axons of the nerve fibers reveal that the secretion in the area of the mantle edge gland is under nervous control. The whole mantle tissue is covered with a columnar epithelium having a microvillar border. The analyses of the outer epithelium show ultrastructural characteristics of a transport active epithelium, indicating that this region of the mantle is involved in the sclerotization of the shell. Ultrastructural findings concerning the epithelium between the outer and middle fold suggest that the periostracum is formed in this area of the mantle, as it is in other conchiferan molluscs. PMID- 15803487 TI - Quantitative evaluation of the fibrin clot extension on different implant surfaces: an in vitro study. AB - The aim of the present study was a quantitative evaluation of the in vitro fibrin clot extension on different implant surfaces. Forty-five disk-shaped commercially pure Grade 2 titanium samples with three different surface topographies (machined, DPS, and Plus) were used in the present study. For the quantitative evaluation of the fibrin clot, 30 specimens were used (10 per group); human whole blood was employed. Venous blood was drawn from three healthy adult volunteers, and 0.2 mL were immediately dropped onto the surface of each specimen. Contact time was 5 min at room temperature; then the samples were rinsed with saline solution and fixed in a buffered solution of glutaraldehyde and paraformaldehyde. Samples were washed again with buffer and dehydrated in an ascending alcohol series. Specimens belonging to all groups were observed under SEM at a magnification of 1000x. From each sample, 50 random micrographs were collected in .tif format with an N x M 1024 x 768 grid of pixels. Quantitative analysis of fibrin clot extension showed the following results: in machined samples fibrin clot extension was 345987.2 +/- 63747.7 pixels(2) (mean +/- SD), in DPS samples fibrin clot extension was 375930.9 +/- 54726.86 pixels(2) (mean +/- SD), and in Plus samples, fibrin clot extension was 612333.6 +/- 46268.42 pixels(2) (mean +/- SD). With ANOVA it was possible to find that there were significant differences among the groups. The Tukey test revealed that the extension of the fibrin clot of Plus samples was statistically higher compared to both machined and DPS samples. The results of this in vitro study indicate that there is a correlation between implant surface morphology and fibrin clot extension. Improvement in surface microtexture complexity seems to determine the formation of a more extensive and three dimensionally complex fibrin scaffold. Further investigations are necessary to explain in more detail the mechanisms that regulate the fibrin clot formation on different implant surfaces. PMID- 15803488 TI - Tissue hyperoxygenation promotes oxidative metabolism in motor unit. AB - Some mutant hemoglobin (Hb) variants are found with lowered O2 affinity. Low oxygen affinity is reported to increase the O2 availability in peripheral tissues (Kunert et al. [1996] Microvasc. Res. 52:58-68). In the present study, we used a mouse model carrying two low-affinity Hb variants, Titusville and Presbyterian, to evaluate the chronic in vivo influence of lowered oxygen affinity on the neuromuscular system. Our model mice showed an increased voluntary running ability compared with wild-type littermates. In the tibialis anterior (TA) muscle of mutant mice, the glycolytic fibers were converted to oxidative ones in where the activity of the mitochondrial marker enzyme succinate dehydrogenase (SDH) was up-regulated. We report that the spinal ventral horn motoneurons innervating TA skeletal fibers also showed higher mitochondrial oxidative enzyme activity. This phenomenon was evidenced by increased SDH activity and electron microscopic (EM) mitochondrial electronic density in these motoneurons. Our data suggest that, as the result of adaptation to the tissue hyperoxygenation, energy metabolism in the neuron-muscle motor unit is augmented and thus function of the motor unit is promoted. PMID- 15803489 TI - Cyto-epitheliochorial placenta of the viviparous lizard Pseudemoia entrecasteauxii: a new placental morphotype. AB - The structural features of the uterine epithelium of the chorioallantoic placenta and omphalloplacenta in the viviparous Australian skink, Pseudemoia entrecasteauxii, were investigated using SEM and TEM techniques. In particular, the structural characteristics that would allow interpretation of function were analyzed, particularly those of gas exchange in the chorioallantoic placenta and histotrophy in the omphaloplacenta. Pseudemoia entrecasteauxii has a complex placenta consisting of a placentome, paraplacentome, and omphaloplacenta. The paraplacentome has a well-vascularized lamina propria in which projecting uterine capillaries displace the overlying uterine epithelial cells, reducing them to attenuated cytoplasmic extensions. Associated cell nuclei and organelles are lost from this region, to provide a capillary lumen to uterine lumen barrier of 0.5 1.0 microm. Hence, the paraplacentome is likely a prominent site for gaseous exchange via simple diffusion. The omphaloplacenta has a similar cytology to that of the placentome, but the uterine epithelial cells are hypertrophied and the apical plasma membrane actively secretes vesicles into the uterine lumen. The omphaloplacenta shows features that are associated with histotrophic transport of nutrients via vesicle secretion, very similar to that of lipid apocrine secretion. The placentome consists of cuboidal cells in the uterine epithelium, with large centrally located nuclei overlying the well-vascularized lamina propria. Although the placentome has a similar cytological structure to that of the omphaloplacenta, granules or active vesicle secretion were not observed. Thus, the placentome may be associated with histotrophy, but not via apocrine secretion. Squamate placentation is epitheliochorial; however, we propose a new term be used to describe the type of placentation in P. entrecasteauxii: "cyto epitheliochorial," because of the extreme attenuation of uterine epithelial cells of the paraplacentome. PMID- 15803490 TI - The role of clofarabine in hematologic and solid malignancies--development of a next-generation nucleoside analog. AB - Clofarabine is a new-generation nucleoside analog that has been synthesized to combine the most favorable pharmacokinetic properties of its congeners fludarabine and cladribine. In addition to inhibition of DNA polymerases and DNA synthesis, clofarabine acts as a strong inhibitor of ribonucleotide reductase (RnR), an enzyme involved in regulating intracellular deoxynucleotide pools, and has a high affinity to the enzyme deoxycytidine kinase (dCyd), the rate-limiting step in nucleoside phosphorylation.A review of the English literature was performed that included original articles and related reviews from the MEDLINE (PubMed) data base and from abstracts based on the publication of meeting materials. Although it was synthesized early in the 1980s, the development of clofarabine was stalled until 1993, when, through efforts at The University of Texas M. D. Anderson Cancer Center, animal toxicology studies were conducted, and the first Phase I study was initiated in patients with hematologic and solid malignancies. Since then, clofarabine has demonstrated single-agent antitumor activity in pediatric and adult acute leukemias. By way of its unique metabolic properties, clofarabine also has lent itself to biochemical modulation strategies with other nucleoside analogs, such as cytarabine. Combinations of clofarabine with cytarabine have been studied in acute leukemia and currently are being evaluated in untreated elderly patients with acute myeloid leukemia. Novel schedules are being explored in lymphoproliferative disorders and solid tumors. Clofarabine is a new nucleoside analog with considerable activity and an acceptable safety profile in acute leukemias. PMID- 15803491 TI - Cytologic features of plasmablastic lymphoma. AB - BACKGROUND: Plasmablastic lymphomas (PBLs) were originally described exclusively in human immunodeficiency virus (HIV)-positive patients who presented with jaw or oral mucosa involvement. Recent studies have reported this neoplasm also in patients without HIV infection and involving sites other than head and neck. This lymphoma has a heterogeneous morphologic presentation but distinct phenotype. METHODS: Cytologic features from four cases of histologically confirmed PBL were evaluated. The cytology specimens were evaluated for criteria as follow: cellularity, cell size and shape, pleomorphism, cytoplasmic characteristics, chromatin pattern, nucleolar features, and mitotic figures. RESULTS: Specimens evaluated were two head and neck fine needle aspiration specimens, one anal smear, and one cerebrospinal fluid specimen. Atypical lymphocytes ranged from intermediate to large in size and demonstrated slight nuclear pleomorphism. The cytoplasm varied from scant to moderate in the alcohol-fixed slides. Nuclei were round with vesicular chromatin. Nucleoli varied from a prominent one to multiple small ones. Multinucleated cells and mitotic figures were easily identified in three of four cases. Tingible-body histiocytes were seen in one case. Ancillary studies in two cases demonstrated expression of CD138 with lack of CD20 expression. CONCLUSION: PBL is a variant of large cell lymphoma with heterogeneous cytologic findings but distinct immunophenotype. Knowledge of the cytomorphologic spectrum of PBLs and detection of CD138 expression by flow cytometry can be helpful in achieving a correct diagnosis. PMID- 15803492 TI - Lomustine (chloroethylnitrosourea [CCNU]), ifosfamide, bleomycin, vincristine, and cisplatin (CIBO-P) is an effective regimen for patients with poor prognostic refractory or multiple disease recurrent aggressive non-Hodgkin lymphoma. AB - BACKGROUND: The current study was designed to assess the activity and safety of a novel combination therapy for patients with recurrent or refractory aggressive non-Hodgkin lymphoma (NHL). METHODS: Forty-three consecutive patients with recurrent or refractory aggressive NHL were treated with lomustine (chloroethylnitrosourea [CCNU]; 60 mg/m2 on Day 1), ifosfamide (1.5 g/m(2 on Days 1, 2 and 21, 22), bleomycin (5 mg/m2 on Days 1, 5 and 21, 25), vincristine (1.4 mg/m2 on Days 1, 8 and 21, 28), and cisplatin (25 mg/m2 on Days 3, 4, 5 and 23, 24, 25), every 42 days (CIBO-P regimen). RESULTS: Thirty-nine patients (91%) were evaluable for response. The median patient age was 63 years. Thirty-five percent of the patients had received > or = 2 lines of previous chemotherapy and 40% had elevated lactate dehydrogenase levels at the time of treatment initiation. The overall objective response rate was 77% (95% confidence interval [95% CI], 63 90%), including 19 (49%) complete (CR) and 11 (28%) partial responses. CIBO-P induced responses in primary refractory disease and in patients treated for second or subsequent disease recurrences. A CR with previous therapy was the most important factor associated with a significantly higher CR rate. The median duration of response was 6 months (95% CI, 4.4-7.7 months) and the median survival duration was 10.7 months (95% CI, 5.9-18.1 months). Five patients (11.6%) remained disease free for > or = 24 months. By multivariate analysis, a CR with previous therapy and average dose intensity of CIBO-P drugs were independent prognostic factors for time-to-treatment failure, whereas a CR with previous therapy and serum lactate dehydrogenase were independent predictors for survival. Myelosuppression was the most frequent serious complication of this regimen. However, none of the patients had hemorrhage with thrombocytopenia, and only 2 patients (5%) had febrile neutropenia. CONCLUSIONS: In the current study, CIBO-P was a novel, highly active, and safe combination therapy for patients with refractory disease with a poor prognosis or for patients with multiply recurrent aggressive NHL. PMID- 15803493 TI - Intraoperative, radio-guided sentinel lymph node mapping in laparoscopic lymph node dissection for Stage I testicular carcinoma. AB - BACKGROUND: The management of regional lymph nodes in patients with clinical Stage I testicular carcinoma is a controversial problem. The authors investigated the feasibility and accuracy of radio-guided mapping of sentinel lymph nodes (SLNs) for men with clinical Stage I testicular tumors. METHODS: Twenty-two patients with clinical Stage I testicular carcinoma were enrolled in the study. One day before surgery, (99m)Technetium-labeled phytate was injected around the testicular tumor. After undergoing radical orchiectomy, patients underwent laparoscopic retroperitoneal lymph node dissection (L-RPLND). All radioactive lymph nodes were marked in the L-RPLND procedure, and three-dimensional SLN maps were made. All resected lymph nodes were evaluated by routine histopathologic examination, and the clinical significance of intraoperative SLN mapping was evaluated. RESULTS: SLNs were detected in 21 of 22 patients (95%). Nearly all SLNs were detected at the ventral or lateral side of the vena cava or at the aorta between the levels of the aortic bifurcation. All SLNs were detected easily in a surgical procedure. Only 1 radio-positive area per patient was identified in 15 patients, and approximately 2-4 positive areas were detected in 6 patients. Two patients had micrometastasis only in SLNs. In 2 patients who had seminoma, lymph node recurrences (at the level of the renal vein and in the obturator lymph node area) occurred at 10 months and 20 months after surgery. CONCLUSIONS: Radio guided mapping of SLNs with laparoscopy was feasible, and nearly all SLNs were detected accurately by the procedure. In the near future, the standard retroperitoneal lymph node dissection may be avoided in most patients with clinical Stage I testicular carcinoma by utilizing focused examination of SLNs. PMID- 15803494 TI - Biotinylated GHK peptide incorporated collagenous matrix: A novel biomaterial for dermal wound healing in rats. AB - Matrikines are small peptide fragments of extracellular matrix proteins that display potent tissue repair activities. Difficulties in achieving sustained delivery of bioactive concentration of matrikines in the affected area limits their therapeutic use. The present study evaluates the effects biotinylated matrikine peptide (bio-glycyl-histidyl-lysine) incorporated collagen membrane for dermal wound healing processes in rats. Biotinylated peptide incorporated collagen matrix (PIC) showed better healing when compared to wounds treated with collagen matrix [CF (collagen film)] and without collagen [CR (control)]. Binding studies indicate that biotinylated GHK (Bio-GHK) binds effectively to the collagen matrix and red blood cell (RBC) membrane when compared with t butyloxycarbonyl substituted GHK (Boc-GHK). Wound contraction, increased cell proliferation, and high expression of antioxidant enzymes in PIC treated group indicate enhanced wound healing activity when compared to CF and CR groups. Interestingly Bio-GHK incorporated collagen increases the copper concentration by ninefold at the wound site indicating the wound healing property of Bio-GHK can also be linked with both copper localization and matrikine activities. These results demonstrate the possibility of using Bio-GHK incorporated collagen film as a therapeutic agent in the wound healing process. PMID- 15803495 TI - Dental adhesive compounds alter glutathione levels but not glutathione redox balance in human THP-1 monocytic cells. AB - The use of hydrophilic dental monomers in dentin bonding agents has vastly improved resin-dentin bond strengths, but incomplete polymerization of these monomers and their leaching into adjacent (pulpal) oral tissues has raised concerns about their biocompatibility. The sublethal effects of these resins are virtually unknown, but their electrophilic nature led to the hypothesis that they may alter cellular oxidative stress pathways. Glutathione balance between reduced (GSH) and oxidized (GSSG) is a major mechanism by which cells maintain redox balance and was therefore the focus of the current investigation. THP-1 human monocytic cells were exposed to hydroxyethyl methacrylate (HEMA), benzoyl peroxide (BPO), camphorquinone (CQ), or triethyelene glycol dimethacrylate (TEGDMA) for 24 h at sublethal doses, then GSH and GSSG levels were measured by means of Ellman's method adapted for cell culture. The results indicate that these dental resin compounds act at least partly via oxidative stress by increasing GSH levels at sublethal concentrations. However, the GSH-GSSG ratio was relatively unaffected. Only BPO altered the GSH-GSSG ratio at 24 h, again at sublethal levels (7.5-15 micromol/L). The results support the hypothesis that resin monomers act, at least in part, via oxidative stress, and that oxidative stress pathways should be one focus of future investigations of monomer biocompatibility. PMID- 15803496 TI - Interaction between LSD and dopamine D2/3 binding sites in pig brain. AB - The psychoactive properties of the hallucinogen LSD have frequently been attributed to high affinity interactions with serotonin 5HT2 receptors in brain. Possible effects of LSD on dopamine D2/3 receptor availability have not previously been investigated in living brain. Therefore, we used PET to map the binding potential (pB) of [11C]raclopride in brain of three pigs, first in a baseline condition, and again at 1 and 4 h after administration of LSD (2.5 microg/kg, i.v.). There was a progressive treatment effect in striatum, where the pB was significantly reduced by 19% at 4 h after LSD administration. Concomitant maps of cerebral blood flow did not reveal significant changes in perfusion during this interval. Subsequent in vitro studies showed that LSD displaced [3H]raclopride (2 nM) from pig brain cryostat sections with an IC50 of 275 nM according to a one-site model. Fitting of a two-site model to the data suggested the presence of a component of the displacement curves with a subnanomolar IC50, comprising 20% of the total [3H]raclopride binding. In microdialysis experiments, LSD at similar and higher doses did not evoke changes in the interstitial concentration of dopamine or its acidic metabolites in rat striatum. Together, these results are consistent with a direct interaction between LSD and a portion of dopamine D2/3 receptors in pig brain, possibly contributing to the psychopharmacology of LSD. PMID- 15803498 TI - Acute effects of physostigmine and galantamine on the binding of [18F]fluoro-A 85380: a PET study in monkeys. AB - 2-[18F]fluoro-3-[2S-2-azetidinylmethoxy]pyridine ([18F]fluoro-A-85380) is an alpha4beta2 subtype selective nicotinic cholinergic agonist with potential suitability for studying changes in endogenous acetylcholine synaptic concentration. Physostigmine, a potent AChE inhibitor, and galantamine, an allosteric modulator of nAChRs, are widely used for the treatment of Alzheimer's disease. Before studying patients with this neurodegenerative disease, positron emission tomography (PET) studies in monkeys were performed to assess the impact of these two compounds on the radiotracer distribution volumes. Physostigmine was administered i.v. at two dosages: 150 microg/kg/h and 37.5 microg/kg/h for 160 min. Galantamine was administered i.v. at two dosages: 2 or 4 mg over 20 min. For PET data analysis, a model with one tissue (radioactivity of the parent compound in plasma and radioactivity in brain tissue) compartment was chosen because reliable parameter estimates could not be obtained with a more complex model. The higher dose of physostigmine produced a 40%, 23%, and 30% reduction of distribution volumes in the putamen, the temporal, and frontal cortices, respectively. The lower dose of physostigmine produced a reduction of 33%, 31%, and 24% in the same structures, respectively. Galantamine (4 mg or 2 mg) produced no significant change of distribution volumes in the basal ganglia, the temporal and frontal cortex. The effects of physostigmine, a more potent AChE inhibitor than galantamine, could be interpreted as a desensitization of nAChRs, due to a prolonged exposure to high synaptic concentration of acetylcholine or as a competition with acetylcholine. PMID- 15803497 TI - Synthesis, characterization, and first successful monkey imaging studies of metabotropic glutamate receptor subtype 5 (mGluR5) PET radiotracers. AB - Three metabotropic glutamate receptor subtype 5 (mGluR5) PET tracers have been labeled with either carbon-11 or fluorine-18 and their in vitro and in vivo behavior in rhesus monkey has been characterized. Each of these tracers share the common features of high affinity for mGluR5 (0.08-0.23 nM vs. rat mGluR5) and moderate lipophilicity (log P 2.8-3.4). Compound 1b was synthesized using a Suzuki or Stille coupling reaction with [11C]MeI. Compounds 2b and 3b were synthesized by a SNAr reaction using a 3-chlorobenzonitrile precursor. Autoradiographic studies in rhesus monkey brain slices using 2b and 3b showed specific binding in cortex, caudate, putamen, amygdala, hippocampus, most thalamic nuclei, and lower binding in the cerebellum. PET imaging studies in monkey showed that all three tracers readily enter the brain and provide an mGluR5-specific signal in all gray matter regions, including the cerebellum. The specific signal observed in the cerebellum was confirmed by the autoradiographic studies and saturation binding experiments that showed tracer binding in the cerebellum of rhesus monkeys. In vitro metabolism studies using the unlabeled compounds showed that 1a, 2a, and 3a are metabolized slower by human liver microsomes than by monkey liver microsomes. In vivo metabolism studies showed 3b to be long-lived in rhesus plasma with only one other more polar metabolite observed. PMID- 15803499 TI - Synaptic differences in the postmortem striatum of subjects with schizophrenia: a stereological ultrastructural analysis. AB - The striatum processes motor, cognitive, and limbic function, all of which are perturbed in schizophrenia. The present study examined the synaptic organization of the caudate and putamen in schizophrenia. Postmortem striatum was obtained from 10 normal controls (NC) and 17 subjects with schizophrenia (SZ), prepared for electron microscopy, and analyzed using stereological principles. The densities of total synapses, asymmetric synapses (characteristic of excitatory inputs), and asymmetric axospinous synapses (characteristic of cortical input) were higher in the caudate of the SZs vs. NCs. These changes were most profound in the off-drug SZ cases and were also elevated in subjects on antipsychotic drugs (APDs). In comparison to NCs, there were no significant differences in the putamen of the SZ cohort as a whole group; however, there were more asymmetric axospinous synapses in the off-drug subgroup. The increase in density of synapses in the SZs does not appear to be caused by antipsychotic medication and may represent failure of normal synaptic pruning or abnormal sprouting. Higher density of cortical-type synapses in SZs vs. NCs may reflect adaptation of corticostriatal circuitry or hyperstimulation of striatal projection neurons. The abnormal synaptic organization could have several important and different downstream effects depending on the precise circuitry involved and may be related to limbic or cognitive dysfunction in schizophrenia. PMID- 15803500 TI - Administration of haloperidol with biperiden reduces mRNAs related to the ubiquitin-proteasome system in mice. AB - In order to find molecules affected by administration of an antipsychotic drug with an antimuscarinic drug, which is a common prescription used to prevent extrapyramidal adverse effects caused by the antipsychotic drugs, gene expression profiling in the frontal cortex was studied in mice. After 14 days of administration with 2 mg/kg haloperidol, a typical antipsychotic drug, and 2 mg/kg biperiden, a high-affinity antagonist for muscarinic receptors in the brain, approximately 500 mRNAs related to synaptic function were investigated. The levels of the mRNAs related to the ubiquitin-related systems were significantly reduced after the combined administration. However, the separate administration of either haloperidol or biperiden had little effect on the levels of the mRNAs. This result suggests that coadministration of haloperidol and biperiden specifically affects the ubiquitin-related system. PMID- 15803501 TI - Functional roles of NMDA receptor NR2A and NR2B subunits in the acute intoxicating effects of ethanol in mice. AB - The present study examined the roles of NR2A and NR2B subunit-containing NMDA receptors in the mediation of the sedative/hypnotic effects of ethanol in mice. The ability of the competitive NMDA antagonist, CGP-37849 (0, 1, or 3 mg/kg), and the NR2B-selective antagonist, Ro 25-6981 (0, 3, or 10 mg/kg), to alter (3 g/kg) ethanol-induced sleep time was measured in C57BL/6J mice and NR2A knockout (KO) mice. The results show that pretreatment with either antagonist significantly potentiated the sedative/hypnotic effects of ethanol in C57BL/6J mice. These effects were not significantly altered in NR2A KO mice. Basal sleep time responses to ethanol were also normal in NR2A KO mice. These findings confirm a major role for NMDA receptors in the acute intoxicating actions of ethanol and provide tentative support for a prepotent role of the NR2B subunit in these effects. PMID- 15803502 TI - BIOLUMBASE--the database of natural and transgenic bioluminescent organisms. AB - The Institute of Biophysics SB RAS hosts and maintains a specialized collection of luminous bacteria (CCIBSO 836) containing over 700 strains isolated in various regions of the world's oceans. The culture collection is a source of lux genes and biologically active substances. The wide application of bioluminescence in medicine and ecology has given importance to analysing information on the structure and functioning of bioluminescence systems in natural and transgenic microorganisms, as well as on their features that are closely interrelated with bioluminescence. The aims of our BIOLUMBASE database are: gathering information on microorganisms with lux genes, their analysis and free access, and distribution of this data throughout the global network. The database includes two sections, natural and transgenic luminous microorganisms, and is updated by our own experimental results, the published literature and internet resources. For the future, a publicly available internet site for BIOLUMBASE is planned. This will list the strains and provide comprehensive information on the properties and functions of luminous bacteria, the mechanisms of regulation of bioluminescence systems, constructs with lux genes, and applications of bioluminescence in microbiology, ecology, medicine and biotechnology. It is noteworthy that this database will also be useful for evaluation of biological hazards of transgenic strains. Users will be able to carry out bibliographic and strain searches starting from any feature of interest. PMID- 15803503 TI - Bibliography. Luminescence literature. GFP and imaging 2002 literature. PMID- 15803504 TI - Bicyclic dioxetanes bearing an inden-2-yl or a benzo(b)thiazol-2-yl moiety as a CIEEL-active chemiluminescent substrate emitting red light. AB - Bicyclic dioxetanes bearing 5-(t-butyldimethylsiloxy)inden-2-yl, 3, or 5-(t butyldimethylsiloxy)benzo(b)thiazol-2-yl, 4, were synthesized. On treatment with large excess of tetrabutylammonium fluoride in DMSO, dioxetane, 3, decomposed rapidly with accompanying emission of red (vermilion) light (lambda(max) (CL) = 637 nm). Comparing the chemiluminescent properties for 3 with those for related dioxetane, 1, in which pi-conjugation system is not fixed in plane, both CIEEL decay rate and chemiluminescent efficiency were found to be improved for 3. Chemiluminescent decomposition of dioxetane, 4, was similarly induced to emit crimson light (lambda(max) (CL) = 725 nm), though the chemiluminescent efficiency was low. PMID- 15803505 TI - Scavenging of reactive oxygen species by the plant phenols genistein and oleuropein. AB - The plant-derived phenolic compounds genistein and oleuropein are known to exhibit several biological properties, many of which may result from their antioxidant and free radical scavenger activity. In this paper we report the results of a complex study of antioxidant activity of genistein and oleuropein, using electron spin resonance (ESR), chemiluminescence, fluorescence and spectrophotometric techniques. Different reaction systems were applied to study the inhibitory effect of the phenolic compounds studied: (a) the potassium superoxide/18-crown-6 dissolved in DMSO system, which generates superoxide radical (O(2).(-)) and hydrogen peroxide (H(2)O(2)); (b) the Co(II)-EDTA-H(2)O(2) system (the Fenton-like reaction), which generates hydroxyl radical (HO.); (c) 2,2'-azobis(2-amidino-propane)dichloride (AAPH) as the peroxyl radical (ROO.) generator, and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical test. Results showed that genistein and oleuropein decreased the chemiluminescence sum from the O(2).(-) generating system, an inhibitory effect that was dependent on their concentration. These compounds also reacted with ROO radicals and they showed activity about two-fold greater than the standard Trolox. The antioxidant effects were studied at different concentrations and reflected in protection against the fluorescence decay of beta-phycoerythrin (beta-PE), due to ROO. attack on this protein. Using the Fenton-like reaction and the spin trap agent 5,5-dimethyl-1 pyrroline-N-oxide (DMPO), the phenolic compounds examined were found to inhibit DMPO-.OH radical formation in the range 10-90% at concentrations of 0.1 mmol/L to 2 mmol/L. Furthermore, these compounds also inhibited HO.-dependent deoxyribose degradation; about 20% and 60% inhibitions were observed in the presence of 0.5 mmol/L genistein and oleuropein, respectively. It was also demonstrated that genistein had a weaker DPPH radical scavenging activity than oleuropein. Our results confirm good scavenging activity towards O(2).(-), HO. and ROO. and the antioxidant effect of genistein and oleuropein. PMID- 15803506 TI - Exuberant thalamocortical axon arborization in cortex-specific NMDAR1 knockout mice. AB - Development of whisker-specific neural patterns in the rodent somatosensory system requires NMDA receptor (NMDAR)-mediated activity. In cortex-specific NR1 knockout (CxNR1KO) mice, while thalamocortical afferents (TCAs) develop rudimentary whisker-specific patterns in the primary somatosensory (barrel) cortex, layer IV cells do not develop barrels or orient their dendrites towards TCAs. To determine the role of postsynaptic NMDARs in presynaptic afferent development and patterning in the barrel cortex, we examined the single TCA arbors in CxNR1KO mice between postnatal days (P) 1-7. Sparsely branched TCAs invade the cortical plate on P1 in CxNR1KO mice as in control mice. In control animals, TCAs progressively elaborate patchy terminals, mostly restricted to layer IV. In CxNR1KO mice, TCAs develop far more extensive arbors between P3-7. Their lateral extent is twice that of controls from P3 onwards. By P7, CxNR1KO TCAs have significantly fewer branch points and terminal endings in layers IV and VI but more in layers II/III and V than control mouse TCAs. Within expansive terminal arbors, CxNR1KO TCAs develop focal terminal densities in layer IV, corresponding to the rudimentary whisker-specific patches. Given that thalamic NMDARs are spared in CxNR1KO mice, the present results show that postsynaptic NMDARs play an important role in refinement of presynaptic afferent arbors and whisker-specific patterning in the developing barrel cortex. PMID- 15803507 TI - Expression and distribution of CB1 cannabinoid receptors in the central nervous system of the African cichlid fish Pelvicachromis pulcher. AB - Neuroanatomical investigation of the cannabinoid system in a lower vertebrate group such as teleost fishes might improve our understanding of the physiological role of such a signaling system. In the present study, the expression of a CB1 cannabinoid receptor has been demonstrated in the CNS of a teleost fish, the cichlid Pelvicachromis pulcher. Moreover, CB1-like immunoreactivity has been analyzed by using a purified antibody against the CB1 receptor amino-terminus. Immunostained neurons and varicosities were found through the telencephalon as well as in the preoptic area and lateral infundibular lobes of the hypothalamus. Stained cells were observed in the pituitary gland. Several cell bodies and nerve terminals containing an intense CB1-like immunoreactivity were found in the pretectal central nucleus and posterior tuberculum, both lying in a transitional region between diencephalon and mesencephalon. In the brainstem, the CB1 immunopositivity was more restricted than in the prosencephalon, with the exception of some large, intensely immunopositive nerve cells within the dorsal mesencephalic tegmentum, possibly motor neurons of the third cranial nerve. In the cerebellum, among a majority of immunonegative granule cells, a subset of them was immunostained. Some positive Purkinje cells were also observed. In the spinal cord, ventral gray matter, several alpha-motoneurons were stained. Similarities to and discrepancies from the CB1 receptor distributions in other vertebrate CNS are discussed, paying particular attention to the abundant CB1 immunoreactivity observed in the area encompassing the pretectum and glomerular nucleus, which is characterized by a peculiar differentiation in bony fishes. PMID- 15803508 TI - The steroid hormone-regulated gene Broad Complex is required for dendritic growth of motoneurons during metamorphosis of Drosophila. AB - Dendrites are subject to subtle modifications as well as extensive remodeling during the assembly and maturation of neural circuits in a wide variety of organisms. During metamorphosis, Drosophila flight motoneurons MN1-MN4 undergo dendritic regression, followed by regrowth, whereas MN5 differentiates de novo (Consoulas et al. [2002] J. Neurosci. 22:4906-4917). Many cellular changes during metamorphosis are triggered and orchestrated by the steroid hormone 20 hydroxyecdysone, which initiates a cascade of coordinated gene expression. Broad Complex (BRC), a primary response gene in the ecdysone cascade, encodes a family of transcription factors (BRC-Z1-Z4) that are essential for metamorphic reorganization of the central nervous system (CNS). Using neuron-filling techniques that reveal cellular morphology with very high resolution, we tested the hypothesis that BRC is required for metamorphic development of MN1-MN5. Through a combination of loss-of-function mutant analyses, genetic mapping, and transgenic rescue experiments, we found that 2Bc function, mediated by BRC-Z3, is required selectively for motoneuron dendritic regrowth (MN1-MN4) and de novo outgrowth (MN5), as well as for soma expansion of MN5. In contrast, larval development and dendritic regression of MN1-MN4 are BRC-independent. Surprisingly, BRC proteins are not expressed in the motoneurons, suggesting that BRC-Z3 exerts its effect in a non-cell-autonomous manner. The 2Bc mutants display no gross defects in overall thoracic CNS structure, or in peripheral structures such as target muscles or sensory neurons. Candidates for mediating the effect of BRC-Z3 on dendritic growth of MN1-MN5 include their synaptic inputs and non neuronal CNS cells that interact with them through direct contact or diffusible factors. PMID- 15803509 TI - Regional abnormalities in retinal development are associated with local ocular hypopigmentation. AB - The retinal pigment epithelium (RPE) plays a key role in regulating retinal development. The critical enzyme in pigment production is tyrosinase. Transgenic mice with a tyrosinase construct where the locus control region was deleted (YRT4) display a variegated phenotype of tyrosinase expression. Their central retina is largely pigment free, whereas more peripheral regions are heavily pigmented. We have used this model to ask whether the influence of pigmented RPE over the retina during development is fundamentally governed by local interactions or is global. Our data show that YRT4 eyes have intermediate melanin content and relatively low tyrosinase activity compared with wild-type and albino animals. Rod counts are comparable to those in pigmented mice in peripheral regions but similar to those in albinos centrally. Anterograde labelling of retinal pathways demonstrates the presence of relatively normal ipsilateral chiasmatic projection in YRT4 mice, comparable with that in pigmented animals and consistent with the peripheral pigmented origin of this pathway. Examination of cellular proliferation levels during retinal development reveals that YRT4 mice display an extended period of mitosis, similar to that found in albinos. Hence, our results show that the regulatory influence of the RPE over the developing retina depends on localized interactions between these tissues. PMID- 15803510 TI - L1 CAM expression in the superficial dorsal horn is derived from the dorsal root ganglion. AB - The cell adhesion molecule L1 is highly expressed on embryonic axons and may play a role in axonal outgrowth and fasciculation. Generally only low levels of L1 are found in adult spinal cord except for intense labeling in Lissauer's tract, in laminae I-II, and on dorsolateral funicular axons. In this study we determine the source of L1 immunoreactivity in the dorsal spinal cord, the presence of L1 expression on sprouting axons, and the effect of exercise on L1 expression. We determined the source of L1 immunoreactivity in the superficial dorsal horn by performing acute unilateral rhizotomies (T12-L4) in adult rats. This resulted in a marked decrease in L1 expression in Lissauer's tract and laminae I-II on the deafferented side. The peptidergic and nonpeptidergic small-diameter primary afferent markers, calcitonin gene-related peptide (CGRP) and the lectin IB4 respectively, closely correlated with L1 expression and also decreased dramatically after rhizotomy. Considering its developmental role, we asked whether L1 was expressed on sprouting axons following chronic rhizotomy. L1 and CGRP, but not IB4, were detected on sprouting axons. Lastly, we investigated the effect of exercise on L1 expression by giving animals with chronic rhizotomies free access to an exercise wheel. After extensive exercise, L1, CGRP, and IB4 expression levels were unchanged compared with those of sedentary chronic animals. Combined, these data demonstrate that the dorsal root ganglia is a major source of L1-positive axons in the superficial dorsal horn, that both L1 and CGRP identify sprouting axons following rhizotomy, and that exercise does not upregulate L1 expression. PMID- 15803512 TI - An algorithm for interpretation of low-energy collision-induced dissociation product ion spectra for de novo sequencing of peptides. AB - An algorithm for interpretation of product ion spectra of peptides generated from ion trap mass spectrometry is developed for de novo amino acid sequencing of peptides for the purpose of protein identification. It is based on a multi-pass analysis of product ion data using a rigorous data extraction and sequence interpretation protocol in the initial pass. The extraction/interpretation algorithm becomes more relaxed in subsequent passes, considering more of the fragment ions, and potentially more sequence candidates. The possible peptide sequences generated by the algorithm are scored according to those sequences which best explain the fragment ion spectrum. These sequences are searched against a protein database using a BLAST search engine to find likely protein candidates. The method is also suitable for locating and determining protein modifications, and can be applied to de novo interpretation of peptide fragment ions in the tandem mass (MS/MS) spectrum produced from a mixture of two peptides having similar nominal mass, but different sequences. Using a known protein, bovine serum albumin, as an example, it is illustrated that this method is rapid and efficient for MS/MS spectral interpretation. This method combined with BLAST programs is then applied to search homologies and to generate information on post translational modifications of an unknown protein isolated from shark cartilage that does not have a complete genome or proteome database. PMID- 15803511 TI - Expression and estrogen-dependent regulation of the zebrafish brain aromatase gene. AB - Compared with adult mammals, the brain of teleost fish is characterized by an extremely high capacity to aromatize androgens into estrogens, and this metabolic activity results from the expression of a specific brain aromatase (AroB) generated by the cyp19b gene. In this study, we first generated antibodies to zebrafish AroB and used them to map AroB-positive structures in the brain of adult zebrafish. We show that AroB is exclusively expressed in radial glial cells, mainly in the olfactory bulbs, telencephalon, preoptic area, and hypothalamus. Second, we investigated in vivo and in vitro the mechanisms involved in the estradiol (E2) regulation of the cyp19b gene. By means of whole mount hybridization and immunohistochemistry on zebrafish embryos and larvae, we confirmed the E2-dependent upregulation of the cyp19b gene, and we show that E2 triggers AroB expression in radial glial cells mainly in the preoptic area and mediobasal hypothalamus of 48 hpf (hours post fertilization) and 108 hpf larvae. In addition, an in vitro analysis of 0.5 kb of the promoter region of the cyp19b gene demonstrated that this E2-dependent regulation involves a direct transcriptional action of estrogen receptors requiring estrogen-responsive elements. However, the data obtained on different cell lines demonstrate that a glial cell context is necessary for full E2 induction. The correlation between our in vivo and in vitro data suggests that the E2-dependent upregulation of AroB is favored by a glial cell context. PMID- 15803514 TI - Electrospray ionization ion trap mass spectrometry for structural characterization of oligosaccharides derivatized with 2-aminobenzamide. AB - The use of electrospray ionization (ESI) quadrupole ion trap mass spectrometry and reversed-phase high-performance liquid chromatography (HPLC) for the characterization of 2-aminobenzamide (2AB)-labeled oligosaccharides and N-linked protein oligosaccharide mixtures is described. The major signals were obtained under these conditions from the [M+Na]+ ions for all 2AB-derivatized oligosaccharides. Under collision-induced dissociation, sodiated molecular species generated in the ESI mode yield simple and predictable mass spectra. Tandem mass spectrometry (MS/MS) experiments with orders higher than two offer a number of ways to enhance MS/MS spectra and to derive information not present in MS and MS2 spectra. Information on composition, sequence, branching and, to some extent, interglycosidic linkages can be deduced from fragments resulting from the cleavage of glycosidic bonds and from weak cross-ring cleavage products. Reversed phase HPLC and derivatization by reductive amination using 2-aminobenzamide were finally applied to characterize a glycan pool enzymatically released from glycoproteins. PMID- 15803513 TI - Site determination of phenyl glycidyl ether-DNA adducts using high-performance liquid chromatography with electrospray ionization tandem mass spectrometry. PMID- 15803515 TI - Species differences in the formation of DA-8164 after intravenous and/or oral administration of DA-8159, a new erectogenic, to mice, rats, rabbits, dogs and humans. AB - Species differences in the formation of DA-8164 after intravenous and/or oral administration of DA-8159 to mice, rats, rabbits, dogs and humans were investigated. After intravenous administration of DA-8159, the formation of DA 8164 decreased in the order mice, rats, rabbits and dogs; the AUC(DA-8164)/AUC(DA 8159) ratios were 0.479, 0.199, 0.0452 and close to 0 (DA-8164 was below the detection limit in dog plasma), respectively. After oral administration of DA 8159, the formation of DA-8164 was considerable in mice, rats and humans, but almost negligible in dogs; the AUC (or AUC(0-t))(DA-8164)/AUC (or AUC(0-t))(DA 8159) ratios were 2.99, 2.67, 1.39 and 0.0650, respectively. The above data suggested that the formation of DA-8164 was almost negligible after both intravenous and oral administration in dogs. The species differences for the formation of DA-8164 may be due to the involvement of different CYP isozymes for each species and/or a different amount or activity of CYP isozyme if the same CYP isozyme is involved for the formation of DA-8164 for all species. The AUC (or AUC(0-t))(DA-8164)/AUC (or AUC(0-t))(DA-8159) ratios after oral administration were greater than those after intravenous administration in mice, rats and dogs, and this could be due to considerable first-pass (gastric, intestinal and/or hepatic) effects in the species as proved in rats. PMID- 15803516 TI - Determination of atomic hydrogen in non-thermal hydrogen plasmas by means of molecular beam threshold ionization mass spectrometry. AB - Atomic hydrogen plays important roles in chemical vapor deposition of functional materials, plasma etching and new approaches to chemical synthesis of hydrogen containing compounds. The present work reports experimental determinations of atomic hydrogen near the grounded electrode in medium-pressure dielectric barrier discharge hydrogen plasmas by means of molecular beam threshold ionization mass spectrometry (MB-TIMS). At certain discharge conditions (a.c. frequency of 24 kHz, 28 kV of peak-to-peak voltage), the measured hydrogen dissociation fraction is decreased from approximately 0.83% to approximately 0.14% as the hydrogen pressure increases from 2.0 to 14.0 Torr. A simulation method for extraction of the approximate electron beam energy distribution function in the mass spectrometer ionizer and a semi-quantitative approach to calibrate the mass discrimination effect caused by the supersonic beam formation and the mass spectrometer measurement are reported. PMID- 15803517 TI - Molecular basis of perhydrolase activity in serine hydrolases. PMID- 15803518 TI - Olefin metatheses and related reactions initiated by carbene derivatives of metals in low oxidation states. AB - Experiments carried out 24 years ago with tantalum carbenes have led to the much cited hypothesis that metals (other than ruthenium) must be in their highest oxidation states for their carbene derivatives to initiate olefin metatheses. The hypothesis legitimizes the uniqueness of high-oxidation-state molybdenum and tungsten carbenes as effective initiators, and it means that the Fischer tungsten carbenes that even earlier were found to initiate olefin metatheses and related transformations must be oxidized before they can be effective. The newer initiators have been termed "well-defined", the older "ill-defined". But what does the evidence show? PMID- 15803519 TI - Bis-chelated imine-alkoxytitanium complexes: novel chiral dopants with high helical twisting power in liquid crystals. AB - Enantiomerically and diastereomerically pure bis-chelated imine-alkoxytitanium complexes 6 and 7 have been synthesized and used as chiral dopants for converting nematic into cholesteric phases. The dopants were tested in mainly commercially available nematic liquid crystalline compounds or mixtures: LC1 (BASF), ZLI-1695 and ZLI-1840 (Merck), as well as N-(4-methoxybenzylidene)-4'-butylaniline (MBBA). The values of the helical twisting power (HTP) were determined by the Grandjean Cano method. Exceptionally high helical twisting powers were obtained. Thus, the titanium complex 6 h displayed a HTP value of 740 microm(-1) in MBBA, the highest HTP value reported. The helical twisting power has been found to depend strongly on the structure of the nematic phase and the substitution pattern of the chiral ligand in the titanium complexes 6 and 7. Crystal structure analysis of 6 f confirmed the A,R,R configuration of the metal complex. The chiral imine ligands 4 and 5 were derived from the regioisomeric amino alcohols 1 and 2. PMID- 15803520 TI - Energy and electron transfer in polyacetylene-linked zinc-porphyrin-[60]fullerene molecular wires. AB - The synthesis and electrochemical and photophysical studies of a series of alkyne linked zinc-porphyrin-[60]fullerene dyads are described. These dyads represent a new class of fully conjugated donor-acceptor systems. An alkynyl-fullerene synthon was synthesized by a nucleophilic addition reaction, and was then oxidatively coupled with a series of alkynyl tetra-aryl zinc-porphyrins with 1-3 alkyne units. Cyclic and differential pulse voltammetry studies confirmed that the porphyrin and fullerene are electronically coupled and that the degree of electronic interaction decreases with increasing length of the alkyne bridge. In toluene, energy transfer from the excited zinc-porphyrin singlet to the fullerene moiety occurs, affording fullerene triplet quantum yields of greater than 90 %. These dyads exhibit very rapid photoinduced electron transfer in tetrahydrofuran (THF) and benzonitrile (PhCN), which is consistent with normal Marcus behavior. Slower rates for charge recombination in THF versus PhCN clearly indicate that charge-recombination events are occurring in the Marcus inverted region. Exceptionally small attenuation factors (beta) of 0.06+/-0.005 A(-1) demonstrate that the triple bond is an effective mediator of electronic interaction in zinc porphyrin-alkyne-fullerene molecular wires. PMID- 15803521 TI - By the way, doctor. I read that glucosamine can affect insulin levels and may be ill-advised for diabetics. Does it cause diabetes in people who don't already have the disease? PMID- 15803522 TI - Health Canada approves rapid HIV test for sale in Canada. PMID- 15803523 TI - Optimal use of propranolol in portal hypertension. PMID- 15803524 TI - [Rehabilitation after knee prostheses]. PMID- 15803525 TI - Share and share alike. PMID- 15803526 TI - Bacteremia among Kenyan Children. PMID- 15803527 TI - Adiposity and physical activity as predictors of mortality. PMID- 15803528 TI - Fecal DNA for colorectal-cancer screening. PMID- 15803530 TI - Medical education in Cuba. PMID- 15803529 TI - Illness and Secrecy on the Supreme Court. PMID- 15803531 TI - Medical education in Cuba. PMID- 15803532 TI - [Pneumogeriatrics and geriatropneumology]. PMID- 15803533 TI - [Respiratory disease in the elderly. A new series of the Revue des Maladies Respiratoires]. PMID- 15803534 TI - [Physiological aspects of the decline of pulmonary function with age]. AB - After peaking between the age of 20 and 30 years, pulmonary function declines gradually with age. This decline is related to changes in respiratory dynamics (lung mechanics, gas exchange) and also to non-respiratory factors (e.g. changes in the immune system). This age-related fall in pulmonary function is not linear, for example there is no further decline in mean PaO2 in men and women, nor in the FEV1/FVC ratio in men, afterthe age of 70 years. Caution is required when interpreting changes in pulmonary mechanics in the elderly due to greater variability of reference values in this age group. PMID- 15803535 TI - [Pharmacokinetics in the elderly]. AB - Physiological changes that occur in the aging process may directly affect drug pharmacokinetics. In the absence of disease or other pathologic conditions, age related changes in pharmacokinetics principally affect drug absorption, distribution, metabolism or elimination. One factor that does change consistently with age is renal clearance, consequently a common pharmacokinetic change is reduced drug clearance, especially for drugs excreted largely unchanged by the kidneys. Alteration of the pharmacokinetics of drugs in the elderly may necessitate adjustment of dosages to prevent toxicity or inadequate therapy. As a broad generalization, dosage should be reduced in elderly patients, reflecting the general decline in body function with age. PMID- 15803536 TI - [Clinical and radiological aspects of ageing of the respiratory tract]. AB - The effects of ageing on the macroscopic appearance of the lung were recognized by Laennec, Andral suggesting that dyspnoea might be its clinical manifestation. At about the same time physiologists demonstrated the negative effects of ageing on the vital capacity, whereas anatomists defined its impact on the lung and thoracic cage structure. The prevalence of dyspnoea increases with ageing but co morbid disease should always be sought. The prevalence of cough is strongly associated with active and passive smoking. With regard to physical signs, barrel chest and crepitations heard over the dependent lung zones do not necessarily have pathological significance. The usefulness of measuring the forced expiratory time remains to be established in the elderly. There is no characteristic radiological feature of a senile lung. If an X-ray abnormality is discovered during a routine examination, it should be regarded as pathological, but in most cases its detection will not alter management of the patient. PMID- 15803537 TI - [Acute dyspnoea in elderly patients]. AB - There is a natural physiological decline in pulmonary function and the cardiovascular system with age. In emergency medicine, acute dyspnoea is a common problem among elderly patients. Some causes, such as pulmonary embolism and diastolic heart failure, are probably under-diagnosed. A good clinical history and examination are as important as arterial blood gas analysis, chest radiography and electrocardiography. Few studies have examined acute dyspnoea in elderly patients, except in the setting of pneumonia. Establishing the underlying diagnosis is often difficult because of atypical presentation and the interaction between cardiac and pulmonary underlying functions. This topic describes several respiratory and cardiac diseases presenting as acute dyspnoea, especially "cardiac asthma" and pulmonary embolism. The clinical usefulness of new investigations such as cardiac and lung echography, pulmonary function tests, serum Brain Natriuretic Peptide and thoracic CT scan are discussed. Further studies looking at acute dyspnoea in elderly patients are needed. PMID- 15803538 TI - [Treatment of lung cancer in the elderly: "Treat them all, God will identify those that belong to Him" or "Requiescat in pace"?]. PMID- 15803539 TI - [Treatment of lung cancer in the elderly]. AB - Elderly patients form an increasing proportion of the lung cancer population. However, they are poorly represented in clinical trials. The published studies are generally phase II trials on highly selected small numbers of patients. Applying these trials to clinical practice is therefore difficult and this is compounded by the fear of increased treatment toxicity in elderly patients. In the event of organ failure (respiratory, cardiac or other organ failure), where conventional therapy is not possible, there are a number of alternative options: radiotherapy for inoperable limited non-small-cell-lung cancer, chemotherapy without platinum for those with more extensive disease or with analogues of platinum for small-cell-lung-cancer. Finally, adjunctive therapy with haematopoietic-cell growth factors or cytoprotectors may allow full doses of treatment to be delivered whilst limiting toxicity. More studies in elderly patients, with larger numbers, are needed to develop more rational therapeutic strategies. We present here some studies of reference and the most recent publications on this subject. PMID- 15803540 TI - [Primary lung cancer in elderly subjects in France]. AB - INTRODUCTION: There are few data on primary bronchial carcinoma in France. We report here the results of the study KBP 2000-CPHG in which there were 1868 patients aged 70 or more and 338 aged 80 or more. METHODS: We compared the patients under 70 (Group I) with those aged 70 or more (Group II) as well as with the details of the over 80's. RESULTS: Group II included significantly more women (17.4% vs 15.2%; p = 0.04), non-smokers (11.2% vs 5.3%; p<0.0001), patients of poor performance status (24.2% vs 14.5%; p<0.0001) and squamous carcinomas (44.5% vs 37.8%; p<0.001) than Group I, but fewer adenocarcinomas (27.2% vs 31.5%; p = 0.009) and as many small cell carcinomas (15.9% vs 16.9%; p = 0.32). In Group II there were less stage III and IV tumours (75.1% vs 78.1%; p = 0.0005) more symptomatic treatment (23.2% vs 6.1%) and radiotherapy alone (12.8% vs 3.8%; p < 0.0001). The results were similar beyond 80 years. On multivariate analysis age, performance status and stage appeared to be independent variables in the choice of curative or symptomatic treatment. CONCLUSIONS: Age alone is not therefore a limiting factor in the choice of treatment. PMID- 15803541 TI - [Immunosenescence and the lung]. AB - INTRODUCTION: Ageing is accompanied by a series of structural and functional changes in the respiratory system, some of which depend on declining performance of the immune system. STATE OF ART: The dominant risk for morbidity and mortality lies in respiratory infections with pneumococci and influenza virus. These risks are increased by immunosenescence, either intrinsic due to ageing or secondary to underlying diseases, poor diet, medication etc. Adaptive responses linked to memory are clearly degraded, particularly the naturally occurring or post vaccination responses to influenza virus antigens or polysaccharide antigens involved in the production of IgG sub class 2 antibodies. Alterations in innate responses have not yet been studied. The role of the immune system associated with the respiratory mucous membranes is reviewed. After the age of 50 several abnormalities develop in the lungs: reduced muco-ciliary clearance, loss of elastic fibres, and low grade inflammation that manifests itself as changes in broncho-alveolar lavage (neutrophilia, increased production of interleukin 8, elastase-antiprotease complexes). One also sees alterations in mononuclear cells with an increase in activated T lymphocytes. The defects of humoral immunity have, in part, been attributed to a functional deficiency of zinc. In the long term low dose replacement together with selenium reduce the frequency and severity of respiratory infections and some of the inadequate responses to vaccination. PERSPECTIVES: The detection and treatment of immune deficiencies associated with ageing are becoming important therapeutic objectives; they may significantly improve the results and clinical benefits of vaccinations. CONCLUSIONS: The declining performance of the immune system with age plays an important role in the development of lung diseases in the elderly. PMID- 15803542 TI - [Non-tubercular respiratory infections in long-term care facilities: advocating for a reinforced prevention]. PMID- 15803543 TI - [Pulmonary infections among the elderly living in institutions: prevention]. AB - Pneumonia amongst elderly people living in institutions is common and is a frequent cause of mortality and hospital admission. It is important to distinguish between prevention of viral pneumonia, which primarily consists of influenza vaccination programmes, and prevention of bacterial pneumonia. Prevention of influenza infection in institutions requires the vaccination of as many as possible of both residents and caregivers. In the event of a declared epidemic then amantadine can be used to reduce the severity of, and complication rate of, influenza infection. The indications for giving this therapy need to be balanced against potential side-effects, especially neurological ones. For the prevention of bacterial pneumonia risk factors such as immobility or impaired swallowing should be first identified and dealt with as necessary. Anti pneumoncoccal vaccination may be considered, but on current evidence, the value of systematic vaccination of residents has not yet been established. PMID- 15803544 TI - [Acute pulmonary infections in elderly patients in institutions]. AB - The high incidence of lower lung infections in elderly subjects in institutions is responsible for an increase in mortality both as the result of the infection itself and of the deterioration of pre-existing conditions. The multiple pathology of these elderly patients living in institutions necessitates particularly detailed diagnosis and management. While the pneumococcus remains the predominant organism the percentage of infections with staphylococci and Gram negative bacteria is higher than among younger subjects. The initial severity should be assessed by clinical and pathological criteria in the same way as the response to treatment. Transfer to hospital is often necessary. Among preventative measures anti-influenzal and anti-pneumococcal vaccines are recommended and reimbursed at 100%. PMID- 15803545 TI - [Tuberculosis in elderly persons living in institutions]. AB - INTRODUCTION: In industrialised countries the incidence of tuberculosis increases with age. STATE OF ART: Tuberculosis remains a current problem in the elderly and may be contracted in an institution. Elderly persons living in institutions account for four times more cases than the elderly living at home. Tuberculosis may present in all forms and situations but the clinical and radiological features may be atypical. Although tuberculin anergy may be found frequently in elderly patients the diagnosis relies on the same principles as in the younger patient. Mortality, even with treatment, is increased and rises with age. Several problems arise in institutions: the screening of sick residents and of subjects at risk as well as the screening of infected patients. PERSPECTIVES: A rigorous policy of prevention and surveillance should lead to a fall in the incidence of the disease in institutions CONCLUSIONS: This collection of data demonstrates that tuberculosis remains a real health problem in elderly subjects in institutions. PMID- 15803546 TI - [Asthma in the elderly]. AB - INTRODUCTION: Asthma in the elderly is a growing clinical problem. It affects 6 to 7% of this age-group, but making the distinction between asthma and chronic obstructive pulmonary disease is more difficult as patients get older. STATE OF THE ART: This difficulty is due to a number of factors: the confounding role of smoking, and also the physiological effects of ageing on the airways which renders airway obstruction more resistant to bronchodilation. Elderly asthmatics can be divided on clinical grounds into two arbitrary groups: "ageing asthmatics" who have had asthma since childhood or adolescence and "late-onset asthmatics" who may present following an infective episode. Certain features of asthma in the elderly include: poor perception of breathlessness, technical difficulties in making reliable pulmonary function measurements and the extrapulmonary manifestations (impact on quality of life). PERSPECTIVES: The therapeutic strategy is much the same as for younger asthmatics but certain aspects take on greater importance: concerns about osteoporosis with long-term corticosteroid therapy (both oral and inhaled), the risk of arrhythmias with beta-2 adrenergic drugs and the significant side-effects of theophylline justifies, in difficult cases, consideration of anticholinergic and/or anti-leukotriene therapy. CONCLUSION: Most importantly, for elderly asthmatics it is a treatment regimen that is as simple as possible and is backed up by a written self-management plan that will improve outcomes. PMID- 15803547 TI - [COPD in elderly patients]. AB - INTRODUCTION: Patients with COPD are growing in number and are getting older, with 75% of deaths occurring after the age of 75; the proportion of women is increasing. STATE OF THE ART: There are few data regarding COPD in the elderly. The related medical and social demands are significant and rise as age and respiratory disability increase. Admission rates to hospital are increasing and, despite the development of services as alternatives to hospital admission, the portion of healthcare resources taken up by elderly COPD patients continues to rise. PERSPECTIVES: Costly therapeutic interventions for COPD should only be employed if they improve the patient's health related quality of life, which is hard to evaluate and correlates poorly with lung function parameters. Although depression is common and more often found in this condition than in others, it cannot be attributed purely to the effects of COPD. CONCLUSION: Preventive measures and new strategies towards more efficient care of elderly patients with COPD must be developed to improve quality of life and to reduce health care costs. PMID- 15803548 TI - [Non-invasive ventilation of elderly patients in the intensive care unit]. AB - INTRODUCTION: An increasing number of elderly patients, with acute respiratory failure (ARF) due to e.g. COPD exacerbation or cardiac failure, are being referred to intensive care units for mechanical ventilation. STATE OF ART: NIV can be an effective ventilatory technique in ARF due to a variety of aetiologies. NIV has been shown to decrease endotracheal intubation rates, complication rates and median hospital length of stay. When managing acute-on-chronic respiratory failure, NIV is a first-line ventilatory technique, in the absence of criteria for immediate endotracheal intubation. The efficacy of NIV depends on the expertise of the medical and nursing staff (including physiotherapists) and should always be performed in an appropriate setting. There are very few data about NIV in the elderly but studies, which included patients over 75 years, did not identify different outcomes for this age group. PERSPECTIVES: Future clinical studies on NIV should allow us to better understand which patients will benefit the most from the technique. Certain specific settings in elderly populations, such as in palliative care or when an "non intubation order" has been given by the patient, his family or the medical/nursing staff, are under evaluation. CONCLUSIONS: NIV is effective and well tolerated and it has become a key ventilatory technique in the management of ARF, particularly for elderly patients. PMID- 15803550 TI - New therapeutic strategies for heart failure: left ventricular remodeling as a target. AB - Structural remodeling of the left ventricle involves myocyte growth and matrix changes that result in chamber enlargement, contractile dysfunction and dyssynchrony. Clinical and experimental data document that these structural changes correlate with progressive worsening of the syndrome of heart failure, that the changes are preventable and reversible, and that a favorable clinical therapeutic response is associated with regression of remodeling. The growing evidence that structural remodeling represents the disease of heart failure has now fueled mechanical as well as pharmacologic approaches to inhibiting the remodeling. PMID- 15803549 TI - [Sleep apnea syndrome in the elderly]. AB - INTRODUCTION: Sleep apnoea/hypopnoea syndrome (SAHS) is distinct in the elderly as age-related co-morbidity may be aggravated by sleep-related breathing disorders or may mask typical clinical symptoms. STATE OF ART: The SAHS prevalence in the elderly varies from one study to another, with an average rate of 25% that might increase according to the importance of associated pathologies. Clinical symptoms tend to be the neurological manifestations of excessive daytime sleepiness and cognitive deterioration. PERSPECTIVES: Treatment of SAHS should include weight reduction strategies if obesity is present. There is no role for surgery in this age group. On the other hand, adjustable oral appliances can be used depending on the severity of the SAHS, but nasal continuous positive airway pressure (CPAP) remains the most effective therapy available. Acceptance of and compliance with CPAP treatment is as good in this age group as in younger patients, with remarkable effectiveness in terms of improvement in daytime sleepiness and cognitive function. However, the effectiveness of treatment for SAHS is related to the level of symptoms present and this should be taken into account when making a therapeutic decision. CONCLUSIONS: These results fully justify treatment of SAHS in the elderly, but the decision to treat should be governed by the level of clinical symptoms from SAHS. PMID- 15803551 TI - Basic mechanisms of left ventricular remodeling: the contribution of wall stress. AB - Heart failure may be viewed as a progressive disorder that is impelled, at least in part, by progressive left ventricular (LV) remodeling. In the present discussion we will review the role of LV remodeling in the pathogenesis of heart failure, with a focus on the contribution that increased wall stress plays in the development and progression of LV remodeling. The clinical implication of this review is that existing neurohormonal strategies may not completely prevent disease progression in the failing heart, and that adjunctive strategies that are designed to specifically prevent or attenuate LV remodeling may play an important role in the clinical treatment of heart failure. PMID- 15803553 TI - Reverse remodeling and enhanced inotropic reserve from the cardiac support device in experimental cardiac failure. AB - Progressive chamber remodeling plays a major role in the pathophysiology of chronic cardiac failure. Recent studies have begun exploring the potential for a passive external containment to impede such progressive dilation. In dogs with ischemic dilated cardiomyopathy, surgical placement of a thin external polyester mesh led to reversal of chronic chamber dilation after 3 to 6 months. Systolic function was preserved compared with the earlier time point. Both end-systolic and end-diastolic chamber volumes were reduced by about 20%, whereas end diastolic pressure and chamber diastolic stiffness were not altered. These findings differ from the natural progression of this model, which involves progressive dilation and systolic dysfunction. In conjunction with reverse remodeling, cardiac inotropic reserve to beta-receptor agonists was markedly enhanced. Furthermore, these changes were induced without adversely affecting Frank-Starling reserve, supporting the lack of constriction. PMID- 15803552 TI - Effects of cardiac support device on reverse remodeling: molecular, biochemical, and structural mechanisms. AB - Progressive left ventricular (LV) dilation is a characteristic feature of heart failure and is invariably associated with poor long-term prognosis. This review discusses observations made in dogs with chronic heart failure using a passive mechanical containment device, the Acorn Cardiac Support Device (CSD), which is designed to prevent progressive LV enlargement. Studies have shown that, in addition to preventing LV dilation, long-term therapy with the CSD also improved LV ejection fraction, tended to normalize LV shape, reduced LV wall stress, and reduced or eliminated functional mitral regurgitation. At the cellular level, the CSD attenuates cardiomyocyte hypertrophy, reduced oxygen diffusion distance and downregulated stretch response proteins. The CSD also improved calcium cycling within the sarcoplasmic reticulum and upregulated mRNA gene expression for alpha myosin heavy chain. These findings, when viewed in concert, provide an explanation for mechanisms that may be responsible for the improvement in LV function seen in dogs with heart after long-term CSD therapy. PMID- 15803554 TI - Surgical therapies for heart failure. AB - A variety of invasive procedures have been utilized to reduce the burden on the left ventricle in order to slow or reverse the progressive changes of structural remodeling. These include mitral valve repair, left ventricular assist devices, left ventricular chamber reduction surgery, endovascular patchplasty, dynamic cardiomyoplasty, and a variety of prosthetic implants designed to inhibit remodeling either by constraining chamber enlargement or reducing wall stress to inhibit further growth. Resynchronization therapy also may favorably affect remodeling. The potential of these procedures to slow the progression of heart failure needs to be confirmed in prospective studies. PMID- 15803555 TI - Worldwide clinical experience with the CorCap Cardiac Support Device. AB - It has been hypothesized that the mechanical burden associated with LV remodeling leads to increased myocardial wall stress and adverse remodeling, all of which serve to further impair cardiac performance and contribute to disease progression. The CorCap Cardiac Support Device (CSD) (Acorn Cardiovascular, Inc., St. Paul, Minnesota) is a mesh-like device that is surgically implanted around the heart. The device is designed to provide circumferential myocardial wall support, and reduce wall stress and myocyte stretch. Clinical experience with CorCap CSD implants in patients with heart failure can be divided into 3 phases: (1) initial safety studies, (2) randomized controlled trials, and (3) limited marketed release. Initial safety studies were undertaken in 48 patients recruited between April 1999 and April of 2001. In 11 patients with complete follow-up, it was noted that LV size, as measured by LV end-diastolic dimension, significantly decreased as early as 3 months postimplant, with an even greater reduction at 6 months. Most importantly, this benefit was sustained, so that the LV end diastolic dimension stayed smaller at 1, 2, and 3 years of follow-up. There was also an improvement in LV function, as manifested by changes in LV ejection fraction. Ejection fraction was significantly increased by 3 months and appeared to reach a peak improvement by 6 months. This benefit was likewise maintained at 1, 2, and 3 years postsurgery. Hemodynamic data did not show any evidence of constrictive physiology. These preliminary safety studies had shown that the CorCap CSD could be implanted safely and without excess operative morbidity or mortality. The primary objective of the Acorn Randomized Clinical Trial is to assess the efficacy and safety of the CorCap CSD in patients with advanced heart failure despite optimal medical therapy. A randomized trial with 2 arms (mitral valve surgery randomized to CSD and cardiomyopathy randomized to medical therapy alone or with CSD) enrolled 300 patients. The primary endpoint of the trial is a change in clinical status from baseline to the end of the efficacy phase as determined by a clinical composite score. The Acorn CorCap CSD received CE mark approval in Europe in 2001 and has been available to a small number of centers in a Limited Market Release Surveillance Study (LMRSS). PMID- 15803556 TI - Cognitive impulsivity and HIV serostatus in substance dependent males. AB - HIV-seropositive (HIV+) drug users show impaired performance on measures of integrity of prefrontal-subcortical systems. The Iowa Gambling Task (GT) is mediated primarily through ventromedial-prefrontal systems, and poor performance on this measure ("cognitive impulsivity") is common among substance dependent individuals (SDIs) as well as patients with disease involving prefrontal subcortical systems (e.g., Huntington disease). We hypothesized that HIV+ SDIs might be more vulnerable to cognitive impulsivity when compared with HIV seronegative (HIV-) SDIs because recent studies report evidence of additive effects of HIV serostatus and drug dependence on cognition. Further, working memory is considered a key component of GT performance and is reliably impaired among HIV+ SDIs compared to controls. We administered the GT to 46 HIV+ and 47 well-matched HIV- males with a past or current history of substance dependence. In addition, we evaluated correlations between subjects' scores on the GT and on a delayed nonmatch to sample (DNMS) task in order to test if working memory deficits accounted for cognitive impulsivity among the HIV+ subjects. The HIV+ subjects performed significantly more poorly on the GT compared to the HIV- group but this effect could not be explained by working memory deficits. Implications of these findings for future basic and applied studies of HIV and substance dependence are discussed. PMID- 15803557 TI - Functional MRI neuroanatomic correlates of the Hooper Visual Organization Test. AB - The Hooper Visual Organization Test (VOT), a commonly applied neuropsychological test of visual spatial ability, is used for assessing patients with suspected right hemisphere, or parietal lobe involvement. A controversy has developed over whether the inferences of this test metric can be assumed to involve global, lateralized, or regional functionality. In this study, the characteristic visual organization and object naming aspects of the VOT task presentation were adapted to a functional MR imaging (fMRI) paradigm to probe the neuroanatomic correlates of this neuropsychological test. Whole brain fMRI mapping results are reported on a cohort of normal subjects. Bilateral fMRI responses were found predominantly in the posterior brain, in regions of superior parietal lobules, ventral temporal occipital cortex, and posterior visual association areas, and to a lesser extent, the frontal eye fields bilaterally, and left dorsolateral prefrontal cortex. The results indicate a general brain region or network in which VOT impairment, due to its visuospatial and object identification demands, is possible to be detected. Discussion is made of interpretive limitations when adapting neuropsychological tests to fMRI analysis. PMID- 15803558 TI - Cognitive impairment in relapsing and primary progressive multiple sclerosis: mostly a matter of speed. AB - Based on the assumption that cognitive impairment in MS is consistent with subcortical dementia, a battery of neuropsychological tests was assembled that included measures of executive function (Tower of London and Wisconsin Card Sorting Test), verbal learning and memory (a paired associates learning test), and speeded information processing (Stroop Color Word Interference Test). The battery was administered to patients with relapsing and primary progressive MS and to healthy controls. Differences between patients and controls occurred on several of the measures. However, when differences with respect to fatigue and depression were statistically controlled, the only differences that remained significant involved measures relating to the speed of information processing. Patients performed more slowly than controls, with the disparity being greater for relapsing patients than for those with primary progressive disease. The slowing was evident on measures of automatic as well as controlled processing and regardless of whether speed was an explicit feature of successful performance or recorded unobstrusively while the patient concentrated on planning a correct solution to a problem. Parallels were noted between cognitive slowing associated with MS and that of normal aging. PMID- 15803559 TI - Neuropsychological and neuropathological effects of anoxic or ischemic induced brain injury. PMID- 15803560 TI - Neuropsychological effects of pediatric obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is a fairly common nocturnal breathing disorder, affecting 2-4% of individuals. Although OSA is associated with medical morbidity, its most functionally disruptive effects in adults appear to be neuropsychological in nature. Research on the neuropsychological effects of pediatric OSA has been limited. This study compared the neuropsychological functioning of school-aged children with OSA to that of healthy children. The primary goal was to clarify the presence and pattern of neuropsychological morbidity associated with pediatric OSA. Sleep was assessed with parent-report questionnaires and laboratory sleep studies. Neuropsychological functioning was assessed by formal tests and parent- and teacher-report questionnaires. Data indicated OSA-related cognitive and behavioral impairment that was particularly marked on measures of behavior regulation and some aspects of attention and executive functioning. Minimal effects were observed on measures of intelligence, verbal memory, or processing speed. Exploratory analyses failed to indicate any clear relationship between neuropsychological functioning and objective indexes of hypoxia or sleep disruption, though the sample was small. These data add to a growing literature which suggests that significant neuropsychological deficits are associated with pediatric OSA. Findings suggest a pattern of neuropsychological morbidity that is similar but not identical to that seen in adult OSA. PMID- 15803561 TI - Attention function after childhood stroke. AB - We investigated attentional outcome after childhood stroke and orthopedic diagnosis in medical controls. Twenty-nine children with focal stroke lesions and individually matched children with clubfoot or scoliosis were studied with standardized attention and neuroimaging assessments. Stroke lesions were quite varied in location and commonly involved regions implicated in Posner's model of attention networks. Children with stroke lesions performed significantly more poorly regarding attention function compared with controls. Performance on the Starry Night, a test demanding alerting and sensory-orienting but not executive attention function, was significantly associated with lesion size in the alerting and sensory-orienting networks but not the executive attention network. Furthermore, earlier age at lesion acquisition was significantly associated with poorer attention function even when lesion size was controlled. These findings support the theory of dissociable networks of attention and add to evidence from studies of children with diffuse and focal brain damage that early insults are associated with worse long-term outcomes in many domains of neuropsychological function. In addition, these results may provide clues towards the understanding of mechanisms underlying attention in children. PMID- 15803562 TI - Long-term neuropsychological outcomes of very low birth weight: associations with early risks for periventricular brain insults. AB - Few follow-up studies of children with very low birth weight (VLBW, <1,500 g) have examined neuropsychological sequelae at later ages or neonatal risks as predictors of these outcomes. The present study assessed cognitive skills at mean age 16 years in 48 participants with <750 g birth weight, 47 with 750-1,499 g birth weight, and 52 term-born controls. Our major objectives were to delineate the long-term cognitive consequences of VLBW, and to determine if risks for periventricular brain insults accounted for variations in outcomes. Analysis revealed poorer outcomes for the <750 g group than for term-born controls on nearly all measures, with specific impairments in visual-motor skills, spatial memory, and executive function. Predictors of outcome for participants with VLBW included lower birth weight, lower weight for gestational age, and a longer period of oxygen requirement for chronic lung disease. The longer-term consequences of VLBW are consistent with expectations based on early brain pathology and suggest limitations to functional plasticity. PMID- 15803564 TI - The role of factor analysis in construct validity: is it a myth? PMID- 15803563 TI - Quality of life, emotional, and cognitive function following acute respiratory distress syndrome. AB - Acute Respiratory Distress Syndrome (ARDS) is characterized by lung injury and hypoxemia, has a high mortality rate, and is associated with significant morbidity including cognitive and emotional sequelae and decreased quality of life. There is limited information regarding which of these factors are associated with decreased quality of life. This study assessed the relationships between quality of life, cognitive and emotional function in ARDS survivors at 1 year post-hospital discharge. Sixty-six ARDS survivors were administered a battery of neuropsychological tests, measures of emotional function and quality of life 1 year post-hospital discharge. At 1 year 45% of the ARDS patients had cognitive sequelae and 29% had mild to moderate symptoms of depression and anxiety. Depression, anxiety, and intensive care unit length of stay were significantly correlated with decreased quality of life. Cognitive impairments did not correlate with decreased quality of life. Illness severity and emotional function, but not cognitive sequelae, are associated with decreased quality of life 1 year following ARDS. ARDS is common and may result in significant cognitive and emotional morbidity and decreased quality of life. PMID- 15803565 TI - [Meningeal central nervous system tuberculosis in children]. PMID- 15803567 TI - Recipe for a healthier diet. New dietary guidelines for a healthy lifestyle: eat more whole grains, fruits, vegetables, and exercise an hour or more a day. PMID- 15803568 TI - Stem cells may cure urinary incontinence. In this experimental treatment, a patient's own cells can stop leaks in as little as one day. PMID- 15803566 TI - Risk marker may play key role in heart disease. PMID- 15803570 TI - Tomato juice may protect against heart disease. PMID- 15803569 TI - The glycemic index method of glucose control. This food-labeling system might be as helpful in maintaining your diabetes as insulin therapy. PMID- 15803571 TI - Poor bone health costs lives. Half of Americans 50 and older may be at risk for fractures. Here's what you can do about it. PMID- 15803572 TI - RA drugs Enbrel, Humira appear safe. PMID- 15803574 TI - Cheap Chinese herb lowers cholesterol. PMID- 15803573 TI - Making coronary bypass surgery last. Your surgeon's decision could influence how soon you need another operation. PMID- 15803575 TI - Chronic stress ages your body. But this mind-body link can be managed. Here's how. PMID- 15803576 TI - Truth or lies: the picture knows. PMID- 15803577 TI - Blocking a key gene could reduce prostate cancer's deadliness. New technology inhibits cancer cell growth. PMID- 15803578 TI - Red wine may lower prostate cancer risk. PMID- 15803579 TI - I'm 68 and worry that I may develop Alzheimer's disease. There's a home test available that can tell you if you may have Alzheimer's based on whether or not you can identify different smells. Does this test work? I think it could give me some peace of mind. PMID- 15803580 TI - Policy and environment assessment on illicit drug use and HIV risk in Cambodia. AB - Research on the extent of illicit drug use and its consequences for HIV risk in Cambodia has revealed that government, NGO, and health care systems in that country are unequipped to deal with HIV/AIDS epidemics among injecting drug users. PMID- 15803581 TI - Technology for safer surgery. PMID- 15803582 TI - OTC defibrillator for cardiac arrest. PMID- 15803583 TI - Noninvasive treatment for uterine fibroids. PMID- 15803584 TI - FDA improves safety of human cells and tissues. PMID- 15803586 TI - Report: prostate cancer, breast cancer leading newly diagnosed cancers. PMID- 15803585 TI - Sentence ordered in drug counterfeiting case. PMID- 15803587 TI - Americans slightly taller, much heavier than 40 years ago. PMID- 15803588 TI - HHS buys 1.2 million doses of flu vaccine. PMID- 15803589 TI - Americans over 50 at risk for bone fractures. PMID- 15803590 TI - A medical militia at the FDA. PMID- 15803591 TI - Safeguards for children taking antidepressants strengthened. PMID- 15803592 TI - Straight talk on braces. PMID- 15803593 TI - Regulatory initiatives for dietary supplements. PMID- 15803594 TI - Use caution buying medical products online. PMID- 15803595 TI - Artificial heart helps people awaiting transplants. PMID- 15803596 TI - Genetics play role in response to asthma drugs. PMID- 15803597 TI - Keeping--or creating--a beautiful smile. PMID- 15803598 TI - Neurological sequelae of chronic profound hypocalcaemia. PMID- 15803599 TI - Breaking bread together. PMID- 15803600 TI - Breaking bread together. PMID- 15803601 TI - Breaking bread together. PMID- 15803602 TI - Space life sciences: search for signatures of life, and space flight environmental effects on the nervous system. PMID- 15803603 TI - Impact-shocked rocks--insights into Archean and extraterrestrial microbial habitats (and sites for prebiotic chemistry?). AB - Impact-shocked gneiss shocked to greater than 10 GPa in the Haughton impact structure in the Canadian High Arctic has an approximately 25-times greater pore surface area than unshocked rocks. These pore spaces provide microhabitats for a diversity of heterotrophic microorganisms and in the near-surface environment of the rocks, where light levels are sufficient, cyanobacteria. Shocked rocks provide a moisture retaining, UV protected microenvironment. During the Archean, when impact fluxes were more than two orders of magnitude higher than today, the shocked-rock habitat was one of the most common terrestrial habitats and might have provided a UV-shielded refugium for primitive life. These potential habitats are in high abundance on Mars where impact crater habitats could have existed over geologic time periods of billions of years, suggesting that impact-shocked rocks are important sites to search for biomolecules in extraterrestrial life detection strategies. In addition to being favourable sites for life, during the prebiotic period of planetary history impact-shocked rocks might have acted as a site for the concentration of reactants for prebiotic syntheses. PMID- 15803605 TI - [Risk factors of viral hepatitis in surgeons]. PMID- 15803604 TI - [Immunotropic treatment of patients with surgical treatment of the gastrointestinal tract]. AB - Results of examination of systemic immunity state, microbiocenosis and immunohistological changes of tissues of the esophagus and stomach in 37 adult patients and 69 children with erosive-ulcerous diseases of the gastrointestinal tract were analyzed. Based on these examinations, up-to-date immunomodulator tamerit inhibiting pathologic activity of monocytes/macrophage system with simultaneous activation of neutrophil granulocytes was used in 154 adult patients and 69 children. Positive result was achieved in all the cases when tamerit was used. PMID- 15803606 TI - [Combined injuries of the neck, thorax, abdomen and cranium]. PMID- 15803608 TI - [Recurrences of varicose disease]. PMID- 15803607 TI - [Policy of surgical treatment in combination of chronic dissection of descending and chronic dissected aneurysm of infrarenal parts of aorta]. PMID- 15803609 TI - [Spontaneous disruption of splenic artery aneurysm as a rare cause of intraabdominal bleeding]. PMID- 15803611 TI - A giant left atrial myxoma prolapsing to midlevel of the left ventricle causing severe pulmonary hypertension. PMID- 15803610 TI - Annual solar UV exposure and biological effective dose rates on the Martian surface. AB - The ultraviolet (UV) environment of Mars has been investigated to gain an understanding of the variation of exposure throughout a Martian year, and link this flux to biological effects and possible survival of organisms at the Martian surface. To gain an idea of how the solar UV radiation varies between different regions, including planned landing sites of two future Mars surface missions, we modelled the total solar UV surface flux throughout one Martian year for two different dust scenarios. To understand the degree of solar UV stress on micro organisms and/or molecules essential for life on the surface of Mars, we also calculated the biologically effective dose (BED) for T7 and Uracil in relevant wavelength regions at the Martian surface as a function of season and latitude, and discuss the biological survival rates in the presence of Martian solar UV radiation. High T7/Uracil BED ratios indicate that even at high latitudes where the UV flux is significantly reduced, the radiation environment is still hostile for life due to the persisting UV-C component of the flux. PMID- 15803612 TI - NSAIDS reduce osteoarthritic knee pain in the short term; long term effects are unknown. PMID- 15803613 TI - Acupuncture does not produce a significant clinical effect in chronic neck pain. PMID- 15803614 TI - 'Usual' physiotherapy is more effective than brief physiotherapy for neck pain. PMID- 15803615 TI - Clinical rule predicts patients likely to benefit from spinal manipulation. PMID- 15803616 TI - Randomisation in clinical trials. PMID- 15803617 TI - Biological space experiments for the simulation of Martian conditions: UV radiation and Martian soil analogues. AB - The survivability of resistant terrestrial microbes, bacterial spores of Bacillus subtilis, was investigated in the BIOPAN facility of the European Space Agency onboard of Russian Earth-orbiting FOTON satellites (BIOPAN I -III missions). The spores were exposed to different subsets of the extreme environmental parameters in space (vacuum, extraterrestrial solar UV, shielding by protecting materials like artificial meteorites). The results of the three space experiments confirmed the deleterious effects of extraterrestrial solar UV radiation which, in contrast to the UV radiation reaching the surface of the Earth, also contains the very energy-rich, short wavelength UVB and UVC radiation. Thin layers of clay, rock or meteorite material were shown to be only successful in UV-shielding, if they are in direct contact with the spores. On Mars the UV radiation climate is similar to that of the early Earth before the development of a protective ozone layer in the atmosphere by the appearance of the first aerobic photosynthetic bacteria. The interference of Martian soil components and the intense and nearly unfiltered Martian solar UV radiation with spores of B. subtilis will be tested with a new BIOPAN experiment, MARSTOX. Different types of Mars soil analogues will be used to determine on one hand their potential toxicity alone or in combination with solar UV (phototoxicity) and on the other hand their UV protection capability. Two sets of samples will be placed under different cut-off filters used to simulate the UV radiation climate of Mars and Earth. After exposure in space the survival of and mutation induction in the spores will be analyzed at the DLR, together with parallel samples from the corresponding ground control experiment performed in the laboratory. This experiment will provide new insights into the principal limits of life and its adaptation to environmental extremes on Earth or other planets which and will also have implications for the potential for the evolution and distribution of life. PMID- 15803618 TI - Current awareness in human psychopharmacology. PMID- 15803619 TI - [Clinical application of kampo medicine for aged patients]. PMID- 15803620 TI - Science, a key AALAS initiative. PMID- 15803621 TI - "Carbon dioxide for euthanasia of laboratory animals". PMID- 15803622 TI - Optimizing treatment of schizophrenia. Enhancing affective/cognitive and depressive functioning. AB - Recognition and treatment of schizophrenia has largely focused on positive symptoms of the disorder, such as delusions, hallucinations, and disorganization. However, other important symptoms, such as depression, cognition, and social functioning, have not received comparable attention. Fifty percent of schizophrenic patients suffer from comorbid depression, which is a major risk factor for suicide in this population, while 10% to 25% suffer from comorbid obsessive-compulsive disorder. Cognitive deficits commonly observed in patients with schizophrenia include problems with concentration, attention, and memory, as well as problem-solving and verbal skills. These deficits are observed at early stages of the illness and can predict deficits in functional capabilities, such as occupational and social skills, educational attainment, and the ability to live independently. The severity of such impairments affects all patient in this population, including up to 10% of patients working full time and up to one third of those working part time. In light of the debilitating effects of depression, cognitive impairment, and other aspects of affective functioning on the quality of life of patients with schizophrenia, physicians need to partner with their patients to address these concerns and determine an appropriate treatment regimen. This can be done with simple functional-based cognitive questioning, the use of evidence-based psychosocial practices, and psychoeducation on the many pharmacotherapeutic options. It is recommended that depressive or suicidal symptoms of schizophrenia be treated with an antidepressant or mood stabilizer only if the symptoms have not subsided after treatment of the psychosis with an atypical antipsychotic. Additionally, relative to older medications, atypicals have demonstrated benefit in improving some of the cognitive impairments. PMID- 15803623 TI - Heavy particle irradiation, neurochemistry and behavior: thresholds, dose response curves and recovery of function. AB - Exposure to heavy particles can affect the functioning of the central nervous system (CNS), particularly the dopaminergic system. In turn, the radiation induced disruption of dopaminergic function affects a variety of behaviors that are dependent upon the integrity of this system, including motor behavior (upper body strength), amphetamine (dopamine)-mediated taste aversion learning, and operant conditioning (fixed-ratio bar pressing). Although the relationships between heavy particle irradiation and the effects of exposure depend, to some extent, upon the specific behavioral or neurochemical endpoint under consideration, a review of the available research leads to the hypothesis that the endpoints mediated by the CNS have certain characteristics in common. These include: (1) a threshold, below which there is no apparent effect; (2) the lack of a dose-response relationship, or an extremely steep dose-response curve, depending on the particular endpoint; and (3) the absence of recovery of function, such that the heavy particle-induced behavioral and neural changes are present when tested up to one year following exposure. The current report reviews the data relevant to the degree to which these characteristics are common to neurochemical and behavioral endpoints that are mediated by the effects of exposure to heavy particles on CNS activity. PMID- 15803624 TI - The effects of proton exposure on neurochemistry and behavior. AB - Future space missions will involve long-term travel beyond the magnetic field of the Earth, where astronauts will be exposed to radiation hazards such as those that arise from galactic cosmic rays. Galactic cosmic rays are composed of protons, alpha particles, and particles of high energy and charge (HZE particles). Research by our group has shown that exposure to HZE particles, primarily 600 MeV/n and 1 GeV/n 56Fe, can produce significant alterations in brain neurochemistry and behavior. However, given that protons can make up a significant portion of the radiation spectrum, it is important to study their effects on neural functioning and on related performance. Therefore, these studies examined the effects of exposure to proton irradiation on neurochemical and behavioral endpoints, including dopaminergic functioning, amphetamine-induced conditioned taste aversion learning, and spatial learning and memory as measured by the Morris water maze. Male Sprague-Dawley rats received a dose of 0, 1.5, 3.0 or 4.0 Gy of 250 MeV protons at Loma Linda University and were tested in the different behavioral tests at various times following exposure. Results showed that there was no effect of proton irradiation at any dose on any of the endpoints measured. Therefore, there is a contrast between the insignificant effects of high dose proton exposure and the dramatic effectiveness of low dose (<0.1 Gy) exposures to 56Fe particles on both neurochemical and behavioral endpoints. PMID- 15803625 TI - The effects of heavy particle irradiation on exploration and response to environmental change. AB - Free radicals produced by exposure to heavy particles have been found to produce motor and cognitive behavioral toxicity effects in rats similar to those found during aging. The present research was designed to investigate the effects of exposure to 56Fe particles on the ability of male Sprague-Dawley rats to detect novel arrangements in a given environment. Using a test of spatial memory previously demonstrated to be sensitive to aging, open field activity and reaction to spatial and non-spatial changes were measured in a group that received a dose of 1.5 Gy (n=10) of 56Fe heavy particle radiation or in non radiated controls (n=10). Animals irradiated with 1.5 Gy of 56Fe particles exhibited some age-like effects in rats tested, even though they were, for the most part, subtle. Animals took longer to enter, visited less and spent significantly less time in the middle and the center portions of the open field, independently of total frequency and duration of activity of both groups. Likewise, irradiated subjects spend significantly more time exploring novel objects placed in the open field than did controls. However, irradiated subjects did not vary from controls in their exploration patterns when objects in the open field were spatially rearranged. Thus, irradiation with a dose of 1.5 Gy of 56Fe high-energy particle radiation elicited age-like effects in general open field exploratory behavior, but did not elicit age-like effects during the spatial and non-spatial rearrangement tasks. PMID- 15803626 TI - Effects of heavy ions on visual function and electrophysiology of rodents: the ALTEA-MICE project. AB - ALTEA-MICE will supplement the ALTEA project on astronauts and provide information on the functional visual impairment possibly induced by heavy ions during prolonged operations in microgravity. Goals of ALTEA-MICE are: (1) to investigate the effects of heavy ions on the visual system of normal and mutant mice with retinal defects; (2) to define reliable experimental conditions for space research; and (3) to develop animal models to study the physiological consequences of space travels on humans. Remotely controlled mouse setup, applied electrophysiological recording methods, remote particle monitoring, and experimental procedures were developed and tested. The project has proved feasible under laboratory-controlled conditions comparable in important aspects to those of astronauts' exposure to particle in space. Experiments are performed at the Brookhaven National Laboratories [BNL] (Upton, NY, USA) and the Gesellschaft fur Schwerionenforschung mbH [GSI]/Biophysik (Darmstadt, FRG) to identify possible electrophysiological changes and/or activation of protective mechanisms in response to pulsed radiation. Offline data analyses are in progress and observations are still anecdotal. Electrophysiological changes after pulsed radiation are within the limits of spontaneous variability under anesthesia, with only indirect evidence of possible retinal/cortical responses. Immunostaining showed changes (e.g. increased expression of FGF2 protein in the outer nuclear layer) suggesting a retinal stress reaction to high-energy particles of potential relevance in space. PMID- 15803627 TI - The ALTEA/ALTEINO projects: studying functional effects of microgravity and cosmic radiation. AB - The ALTEA project investigates the risks of functional brain damage induced by particle radiation in space. A modular facility (the ALTEA facility) is being implemented and will be operated in the International Space Station (ISS) to record electrophysiological and behavioral descriptors of brain function and to monitor their time dynamics and correlation with particles and space environment. The focus of the program will be on abnormal visual perceptions (often reported as "light flashes" by astronauts) and the impact on retinal and brain visual structures of particle in microgravity conditions. The facility will be made available to the international scientific community for human neurophysiological, electrophysiological and psychophysics experiments, studies on particle fluxes, and dosimetry. A precursor of ALTEA (the 'Alteino' project) helps set the experimental baseline for the ALTEA experiments, while providing novel information on the radiation environment onboard the ISS and on the brain electrophysiology of the astronauts during orbital flights. Alteino was flown to the ISS on the Soyuz TM34 as part of mission Marco Polo. Controlled ground experiments using mice and accelerator beams complete the experimental strategy of ALTEA. We present here the status of progress of the ALTEA project and preliminary results of the Alteino study on brain dynamics, particle fluxes and abnormal visual perceptions. PMID- 15803628 TI - Exposure of animals to artificial gravity conditions leads to the alteration of the glutamate release from rat cerebral hemispheres nerve terminals. AB - The biochemical basis underlying the effects of altered gravity on the process of nervous signal transmission is not clear. We have investigated the effect of hypergravity stress (created by centrifugation of rats at l0 g for 1 h) on the basal and stimulated release of L-[14C]glutamate (a chemical transmitter of excitatory signals) from isolated rat brain nerve terminals (synaptosomes). It has been shown that the hypergravity stress exerted a different influence on the Ca(2+)-dependent and the Ca(2+)-independent component of neurotransmitter release. The Ca(2+)-dependent L-[14C]glutamate release evoked by potassium chloride was equal to 14.4 +/- 0.7% of total synaptosomal label for control animals and 6.2 +/- 1.9% for animals, exposed to hypergravity (P < or = 0.05) and was more than twice decreased as a result of the hypergravity stress. We observed no statistically significant difference in the Ca(2+)-independent component of L [14C]glutamate release. For control group and animals exposed to the hypergravity stress it was equal to 7.7 +/- 2.8% and 12.9 +/- 2.0%, respectively. We have also investigated the effect of the hypergravity stress on the activity of high affinity Na(+)-dependent glutamate transporters. Km and Vmax of L-[14C]glutamate uptake have been determined. The maximal velocity of glutamate uptake was decreased as a result of hypergravity loading, but no difference in the Km values between control rats and hypergravity exposed animals was observed. These findings indicate that hypergravity stress alters neurotransmitter reuptake and exocytotic neurotransmitter release processes. PMID- 15803629 TI - Short term microgravity effect on isometric hand grip and precision pinch force with visual and proprioceptive feedback. AB - Experiments executed on the upper limb are assuming increasing significance in the frame of the Human Physiology in space, for at least two reasons: the upper limb is the principal means of locomotion for the subject living in a space station; furthermore, fatigue can have a significant effect on the hand, for the ordinary work on board, and in particular for the extra-vehicular activities. The degradation of the performances affecting the muscular-skeletal apparatus can be easily recognized on the upper limb, by exerting specific scientific protocols, to be repeated through the permanence of the subject in weightlessness conditions. Another aspect relevant to the effect of microgravity on the upper limb is associated with the alteration of the motor control programs due to the different gravity factor, affecting not only the bio-mechanics of the subject, but in general all his/her psycho-physical conditions, induced by the totally different environment. Specific protocols on the upper limb can facilitate the studies on learning mechanisms for the motor control. The results of such experiments can be transferred to the Earth, useful for treatment of subjects with local traumas or diseases of the Central Nervous System. PMID- 15803630 TI - CNS development under altered gravity: cerebellar glial and neuronal protein expression in rat neonates exposed to hypergravity. AB - The future of space exploration depends on a solid understanding of the developmental process under microgravity, specifically in relation to the central nervous system (CNS). We have previously employed a hypergravity paradigm to assess the impact of altered gravity on the developing rat cerebellum. The present study addresses the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of selected glial and neuronal cerebellar proteins in rat neonates exposed to hypergravity (1.5 G) from embryonic day (E)11 to postnatal day (P)6 or P9 (the time of maximal cerebellar changes) comparing them against their expression in rat neonates developing under normal gravity. Proteins were analyzed by quantitative Western blots of cerebellar homogenates; RNA analysis was performed in the same samples using quantitative PCR. Densitometric analysis of Western blots suggested a reduction in glial (glial acidic protein, GFAP) and neuronal (neuronal cell adhesion molecule, NCAM-L1, synaptophysin) proteins, but the changes in individual cerebellar proteins in hypergravity-exposed neonates appeared both age- and gender-specific. RNA analysis suggested a reduction in GFAP and synaptophysin mRNAs on P6. These data suggest that exposure to hypergravity may interfere with the expression of selected cerebellar proteins. These changes in protein expression may be involved in mediating the effect of hypergravity on the developing rat cerebellum. PMID- 15803631 TI - International Conference on Harmonisation; guidance on E2E Pharmacovigilance Planning; availability. Notice. AB - The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled "E2E Pharmacovigilance Planning." The guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The guidance describes a method for summarizing the important potential and identified risks of a drug. It proposes a structure for a pharmacovigilance plan and sets out principles of good practice for the design and conduct of observational studies. The guidance is intended to aid in planning pharmacovigilance activities, especially in preparation for the early postmarketing period of a new drug. PMID- 15803632 TI - Fitness to work of astronauts in conditions of action of the extreme emotional factors. AB - The theoretical model for the quantitative determination of influence of a level of emotional exertion on the success of human activity is presented. The learning curves of fixed words in the groups with a different level of the emotional exertion are analyzed. The obtained magnitudes of time constant T depending on a type of the emotional exertion are a quantitative measure of the emotional exertion. Time constants could also be of use for a prediction of the characteristic of fitness to work of an astronaut in conditions of extreme factors. The inverse of the sign of influencing on efficiency of activity of the man is detected. The paper offers a mathematical model of the relation between successful activity and motivations or the emotional exertion (Yerkes-Dodson law). Proposed models can serve by the theoretical basis of the quantitative characteristics of an estimation of activity of astronauts in conditions of the emotional factors at a phase of their selection. PMID- 15803633 TI - Effect of hypergravity on carboanhydrase reactivity in inner ear ionocytes of developing cichlid fish. AB - It has been shown earlier that hypergravity slows down inner ear otolith growth in developing fish. Otolith growth in terms of mineralization mainly depends on the enzyme carboanhydrase (CA), which is responsible for the provision of the pH value necessary for calcium carbonate deposition. Larval siblings of cichlid fish (Oreochromis mossambicus) were subjected to hypergravity (3 g, hg; 6 h) during development and separated into normally and kinetotically swimming individuals following the transfer to 1 g (i.e., stopping the centrifuge; kinetotically behaving fish performed spinning movements). Subsequently, CA was histochemically demonstrated in inner ear ionocytes (cells involved in the endolymphatic ion exchange) and enzyme reactivity was determined densitometrically. It was found that both the total macular CA-reactivity as well as the difference in reactivities between the left and the right maculae (asymmetry) were significantly lower (1) in experimental animals as compared to the 1 g controls and (2) in normally swimming hg-animals as compared to the kinetotically behaving hg-fish. The results are in complete agreement with earlier studies, according to which hypergravity induces a decrease of otolith growth and the otolithic calcium incorporation (visualized using the calcium-tracer alizarin complexone) of kinetotically swimming hg-fish was higher as compared to normally behaving hyper g animals. The present study thus strongly supports the concept that a regulatory mechanism, which adjusts otolith size and asymmetry as well as otolithic calcium carbonate incorporation towards the gravity vector, acts via activation/deactivation of macular CA. PMID- 15803634 TI - Swimming behaviour and calcium incorporation into inner ear otoliths of fish after vestibular nerve transection. AB - Previous investigations on neonate swordtail fish (Xiphophorus helleri) revealed that otolithic calcium incorporation (visualized using the calcium tracer alizarin complexone) and thus otolith growth had ceased after nerve transection, supporting a hypothesis according to which the gravity-dependent otolith growth is regulated neuronally. Subsequent investigations on larval cichlid fish (Oreochromis mossambicus) yielded contrasting results, repeatedly depending on the particular batch of cichlids investigated. Like most neonate swordtails, Type I cichlids revealed a stop of calcium incorporation after unilateral vestibular nerve transection. Their behaviour after transection was normal, and the otolithic calcium incorporation in controls of the same batch was symmetric. In Type II cichlids, however, vestibular nerve transection had no effect on otolithic calcium incorporation. They behaved kinetotically after transection (this kind of kinetosis was qualitatively similar to the swimming behaviour exhibited by larval cichlids during microgravity in the course of parabolic aircraft flights). The otolithic calcium incorporation in control animals was asymmetric. These results show that the effects of vestibular nerve transection as well as the efficacy of the mechanism, which regulates otolith growth/otolithic calcium incorporation, are--depending on the particular batch of animals--genetically predispositioned. In conclusion, the regulation of otolithic calcium incorporation is guided neuronally, in part via the vestibular nerve and, in part, via a further pathway, which remains to be addressed in the course of future investigations. PMID- 15803635 TI - Calcium gradients in the fish inner ear sensory epithelium and otolithic membrane visualized by energy filtering transmission electron microscopy (EFTEM). AB - Inner ear otolith formation in fish is supposed to be performed by the molecular release of proteinacious precursor material from the sensory epithelia, followed by an undirected and diffuse precipitation of calcium carbonate (which is mainly responsible for the functionally important weight of otoliths). The pathway of calcium into the endolymph, however, still remains obscure. Therefore, the presence of calcium within the utricle of larval cichlid fish Oreochromis mossambicus was analyzed by means of energy filtering transmission electron microscopy (EFTEM). Electron spectroscopic imaging (ESI) and electron energy loss spectra (EELS) revealed discrete calcium precipitations, which were especially numerous in the proximal endolymph as compared to the distal endolymph. A decreasing proximo-distal gradient was also present within the proximal endolymph between the sensory epithelium and the otolith. Further calcium particles covered the peripheral proteinacious layer of the otolith. They were especially pronounced at the proximal surface of the otolith. Other calcium precipitates were found to be accumulated at the macular junctions. These results strongly suggest that the apical region of the macular epithelium is involved in the release of calcium and that calcium supply of the otoliths takes place in the proximal endolymph. PMID- 15803636 TI - Extremely elongated mitochondria in ionocytes of the saccular epithelium of a teleost, Oreochromis mossambicus (Cichlidae). AB - Unusually large mitochondria are a rather scarce feature in normal biological tissue and string-like giant mitochondria have hitherto not been reported in animals. Investigating the role of inner ear ionocytes for otolith growth, large ionocytes of the saccular epithelium of the cichlid fish Oreochromis mossambicus were analyzed by imaging of thick sections with energy-filtering transmission electron microscopy. We report here that ionocytes do not contain numerous small sized mitochondria as has been suggested earlier but rather few, extremely elongated megamitochondria. Since the particular mitochondrial structure is important for normal cell function, such megamitochondria possibly reflect a functional advantage in the context of the presumed role of teleostean ionocytes in regulating the composition of the endolymphatic fluid. PMID- 15803637 TI - No correlation between multilamellar bodies in the inner ear and further organs of mutant (backstroke, bks) and wildtype zebrafish embryos. AB - The origin of the proteinacious matrix of the inner ear stones (otoliths) of vertebrates has not yet been clarified. Using the backstroke mutant (bks) of the zebrafish Danio rerio, which is characterized by a complete lack of otoliths, we searched for possibly missing or aberrant structural components within the macular epithelia of the inner ears of embryos on the ultrastructural level. Numerous multilamellar bodies (MLBs) were found. The MLBs were, however, not restricted to the inner ears of mutants but were also found in wildtype individuals and in further organs such as brain and liver. MLBs have hitherto never been described from the inner ear of fish and are generally estimated to be rare structures. Their occurrence in fish liver can, however, be induced by using particular chemical substances, which seem to effect adaptive compensatory processes on the cellular level. Such a chemical treatment also affects the ultrastructure of further organelles. Since the occurrence of MLBs in the liver of zebrafish was not accompanied by an alteration of the morphology of other organelles, their occurrence seems not to be due to environmental stress. The findings indicate that the MLBs cannot be correlated with bks-inherent features as well as with missing otolith development/growth. Since the occurrence of MLBs was independent from the developmental stage of a specimen and its overall tissue preservation, it can moreover be excluded that these MLBs merely represent fixation artifacts. Their presence more likely indicates cellular remodelling processes of hitherto unknown significance. PMID- 15803638 TI - [Color Doppler ultrasonographic diagnosis of fetal omphalocele: report of two cases]. PMID- 15803639 TI - [Uterine necrosis, abdominal cavity infection, and extensive colon necrosis due to endometrial clear-cell carcinoma: report of one case]. PMID- 15803640 TI - [Retroperitoneal paraganglioma: report of 2 cases]. PMID- 15803641 TI - [Duodenal tumor complicated by tachycardia: report of one case]. PMID- 15803642 TI - [Complication after implantable cardioverter defibrillator implantation: report of 1 case]. PMID- 15803643 TI - [Memorable dates and anniversaries in history of clinical medicine in 2005]. PMID- 15803644 TI - [Special form of wellness in Austria]. PMID- 15803645 TI - Perspectives on biotechnological applications of archaea. AB - Many archaea colonize extreme environments. They include hyperthermophiles, sulfur-metabolizing thermophiles, extreme halophiles and methanogens. Because extremophilic microorganisms have unusual properties, they are a potentially valuable resource in the development of novel biotechnological processes. Despite extensive research, however, there are few existing industrial applications of either archaeal biomass or archaeal enzymes. This review summarizes current knowledge about the biotechnological uses of archaea and archaeal enzymes with special attention to potential applications that are the subject of current experimental evaluation. Topics covered include cultivation methods, recent achievements in genomics, which are of key importance for the development of new biotechnological tools, and the application of wild-type biomasses, engineered microorganisms, enzymes and specific metabolites in particular bioprocesses of industrial interest. PMID- 15803646 TI - The Sso7d protein of Sulfolobus solfataricus: in vitro relationship among different activities. AB - The physiological role of the nonspecific DNA-binding protein Sso7d from the crenarchaeon Sulfolobus solfataricus is unknown. In vitro studies have shown that Sso7d promotes annealing of complementary DNA strands (Guagliardi et al. 1997), induces negative supercoiling (Lopez-Garcia et al. 1998), and chaperones the disassembly and renaturation of protein aggregates in an ATP hydrolysis-dependent manner (Guagliardi et al. 2000). In this study, we examined the relationships among the binding of Sso7d to double-stranded DNA, its interaction with protein aggregates, and its ATPase activity. Experiments with 1-anilinonaphthalene-8 sulfonic acid as probe demonstrated that exposed hydrophobic surfaces in Sso7d are responsible for interactions with protein aggregates and double-stranded DNA, whereas the site of ATPase activity has a non-hydrophobic character. The interactions of Sso7d with double-stranded DNA and with protein aggregates are mutually exclusive events, suggesting that the disassembly activity and the DNA related activities of Sso7d may be competitive in vivo. In contrast, the hydrolysis of ATP by Sso7d is independent of the binding of Sso7d to double stranded DNA or protein aggregates. PMID- 15803647 TI - Cellular localization of D-lactate dehydrogenase and NADH oxidase from Archaeoglobus fulgidus. AB - Members of the genus Archaeoglobus are hyperthermophilic sulfate reducers with an optimal growth temperature of 83 degrees C. Archaeoglobus fulgidus can utilize simple compounds including D-lactate, L-lactate and pyruvate as the sole substrate for carbon and electrons for dissimilatory sulfate reduction. Previously we showed that this organism makes a D-lactate dehydrogenase (Dld) that requires FAD and Zn2+ for activity. To determine the cellular location and topology of Dld and to identify proteins that interact with Dld, an antibody directed against Dld was prepared. Immunocytochemical studies using gold particle coated secondary antibodies show that more than 85% of Dld is associated with the membrane. A truncated form of Dld was detected in immunoblots of whole cells treated with protease, showing that Dld is an integral membrane protein and that a significant portion of Dld, including part of the FAD-binding pocket, is outside the membrane facing the S-layer. The gene encoding Dld is part of an operon that includes noxA2, which encodes one of several NADH oxidases in A. fulgidus. Previous studies have shown that NoxA2 remains bound to Dld during purification. Thin sections of A. fulgidus probed simultaneously with antibodies against Dld and NoxA2 show that both proteins co-localized to the same sites in the membrane. Although these data show a tight interaction between NoxA2 and Dld, the role of NoxA2 in electron transport reactions is unknown. Rather, NoxA2 may protect proteins involved in electron transfer by reducing O2 to H2O2 or H2O. PMID- 15803648 TI - Reverse micelles in organic solvents: a medium for the biotechnological use of extreme halophilic enzymes at low salt concentration. AB - Alkaline p-nitrophenylphosphate phosphatase (pNPPase) from the halophilic archaeobacterium Halobacterium salinarum (previously halobium) was solubilized at low salt concentration in reverse micelles of hexadecyltrimethyl-ammoniumbromide in cyclohexane with 1-butanol as co-surfactant. The enzyme maintained its catalytic properties under these conditions. The thermodynamic "solvation stabilization hypothesis" has been used to explain the bell-shaped dependence of pNPPase activity on the water content of reverse micelles, in terms of protein solvent interactions. According to this model, the stability of the folded protein depends on a network of hydrated ions associated with acidic residues at the protein surface. At low salt concentration and low water content (the ratio of water concentration to surfactant concentration; w0), the network of hydrated ions within the reverse micelles may involve the cationic heads of the surfactant. The bell-shaped profile of the relationship between enzyme activity and w0 varied depending on the concentrations of NaCl and Mn2+. PMID- 15803649 TI - Pyrobaculum calidifontis sp. nov., a novel hyperthermophilic archaeon that grows in atmospheric air. AB - A novel, facultatively aerobic, heterotrophic hyperthermophilic archaeon was isolated from a terrestrial hot spring in the Philippines. Cells of the new isolate, strain VA1, were rod-shaped with a length of 1.5 to 10 microm and a width of 0.5 to 1.0 microm. Isolate VA1 grew optimally at 90 to 95 degrees C and pH 7.0 in atmospheric air. Oxygen served as a final electron acceptor under aerobic growth conditions, and vigorous shaking of the medium significantly enhanced growth. Elemental sulfur inhibited cell growth under aerobic growth conditions, whereas thiosulfate stimulated cell growth. Under anaerobic growth conditions, nitrate served as a final electron acceptor, but nitrite or sulfur containing compounds such as elemental sulfur, thiosulfate, sulfate and sulfite could not act as final electron acceptors. The G+C content of the genomic DNA was 51 mol%. Phylogenetic analysis based on 16S rRNA sequences indicated that strain VA1 exhibited close relationships to species of the genus Pyrobaculum. A DNA-DNA hybridization study revealed a low level of similarity (< or = 18%) between strain VA1 and previously described members of the genus Pyrobaculum. Physiological characteristics also indicated that strain VA1 was distinct from these Pyrobaculum species. Our results indicate that isolate VA1 represents a novel species, named Pyrobaculum calidifontis. PMID- 15803650 TI - Structural analysis by reductive cleavage with LiAlH4 of an allyl ether choline phospholipid, archaetidylcholine, from the hyperthermophilic methanoarchaeon Methanopyrus kandleri. AB - A choline-containing phospholipid (PL-4) in Methanopyrus kandleri cells was identified as archaetidylcholine, which has been described by Sprott et al. (1997). The PL-4 consisted of a variety of molecular species differing in hydrocarbon composition. Most of the PL-4 was acid-labile because of its allyl ether bond. The identity of PL-4 was confirmed by thin-layer chromatography followed by positive staining with Dragendorff reagent and fast-atom bombardment mass spectrometry. A new method of LiAlH4 hydrogenolysis was developed to cleave allyl ether bonds and recover the corresponding hydrocarbons. We confirmed the validity of the LiAlH4 method in a study of the model compound synthetic unsaturated archaetidic acid (2,3-di-O-geranylgeranyl-sn-glycerol-1-phosphate). Saturated ether bonds were not cleaved by the LiAlH4 method. The hydrocarbons formed following LiAlH4 hydrogenolysis of PL-4 were identified by gas-liquid chromatography and mass spectrometry. Four kinds of hydrocarbons with one to four double bonds were detected: 47% of the hydrocarbons had four double bonds; 11% had three double bonds; 14% had two double bonds; 7% had one double bond; and 6% were saturated species. The molecular species composition of PL-4 was also estimated based on acid lability: 77% of the molecular species had two acid labile hydrocarbons; 11% had one acid-labile and one acid-stable hydrocarbon; and 11% had two acid-stable hydrocarbons. To our knowledge, this is the first report of a specific chemical degradation method for the structural analysis of allyl ether phospholipid in archaea. PMID- 15803651 TI - Molecular analysis of the role of two aromatic aminotransferases and a broad specificity aspartate aminotransferase in the aromatic amino acid metabolism of Pyrococcus furiosus. AB - The genes encoding aromatic aminotransferase II (AroAT II) and aspartate aminotransferase (AspAT) from Pyrococcus furiosus have been identified, expressed in Escherichia coli and the recombinant proteins characterized. The AroAT II enzyme was specific for the transamination reaction of the aromatic amino acids, and uses a-ketoglutarate as the amino acceptor. Like the previously characterized AroAT I, AroAT II has highest efficiency for phenylalanine (k(cat)/Km = 923 s(-1) mM(-1)). Northern blot analyses revealed that AroAT I was mainly expressed when tryptone was the primary carbon and energy source. Although the expression was significantly lower, a similar trend was observed for AroAT II. These observations suggest that both AroATs are involved in amino acid degradation. Although AspAT exhibited highest activity with aspartate and alpha-ketoglutarate (k(cat) approximately 105 s(-1)), it also showed significant activity with alanine, glutamate and the aromatic amino acids. With aspartate as the amino donor, AspAT catalyzed the amination of alpha-ketoglutarate, pyruvate and phenyl pyruvate. No activity was detected with either branched-chain amino acids or alpha-keto acids. The AspAT gene (aspC) was expressed as a polycistronic message as part of the aro operon, with expression observed only when the aromatic amino acids were absent from the growth medium, indicating a role in the biosynthesis of the aromatic amino acids. PMID- 15803652 TI - Functional organization of a single nif cluster in the mesophilic archaeon Methanosarcina mazei strain Go1. AB - The mesophilic methanogenic archaeon Methanosarcina mazei strain Go1 is able to utilize molecular nitrogen (N2) as its sole nitrogen source. We have identified and characterized a single nitrogen fixation (nif) gene cluster in M. mazei Go1 with an approximate length of 9 kbp. Sequence analysis revealed seven genes with sequence similarities to nifH, nifI1, nifI2, nifD, nifK, nifE and nifN, similar to other diazotrophic methanogens and certain bacteria such as Clostridium acetobutylicum, with the two glnB-like genes (nifI1 and nifI2) located between nifH and nifD. Phylogenetic analysis of deduced amino acid sequences for the nitrogenase structural genes of M. mazei Go1 showed that they are most closely related to Methanosarcina barkeri nif2 genes, and also closely resemble those for the corresponding nif products of the gram-positive bacterium C. acetobutylicum. Northern blot analysis and reverse transcription PCR analysis demonstrated that the M. mazei nif genes constitute an operon transcribed only under nitrogen starvation as a single 8 kb transcript. Sequence analysis revealed a palindromic sequence at the transcriptional start site in front of the M. mazei nifH gene, which may have a function in transcriptional regulation of the nif operon. PMID- 15803654 TI - The ultrastructure of Ignicoccus: evidence for a novel outer membrane and for intracellular vesicle budding in an archaeon. AB - A novel genus of hyperthermophilic, strictly chemolithotrophic archaea, Ignicoccus, has been described recently, with (so far) three isolates in pure culture. Cells were prepared for ultrastructural investigation by cultivation in cellulose capillaries and processing by high-pressure freezing, freeze substitution and embedding in Epon. Cells prepared in accordance with this protocol consistently showed a novel cell envelope structure previously unknown among the Archaea: a cytoplasmic membrane; a periplasmic space with a variable width of 20 to 400 nm, containing membrane-bound vesicles; and an outer sheath, approximately 10 nm wide, resembling the outer membrane of gram-negative bacteria. This sheath contained three types of particles: numerous tightly, irregularly packed single particles, about 8 nm in diameter; pores with a diameter of 24 nm, surrounded by tiny particles, arranged in a ring with a diameter of 130 nm; and clusters of up to eight particles, each particle 12 nm in diameter. Freeze-etched cells exhibited a smooth surface, without a regular pattern, with frequent fracture planes through the outer sheath, indicating the presence of an outer membrane and the absence of an S-layer. The study illustrates the novel complex architecture of the cell envelope of Ignicoccus as well as the importance of elaborate preparation procedures for ultrastructural investigations. PMID- 15803653 TI - Identification and analysis of proton-translocating pyrophosphatases in the methanogenic archaeon Methansarcina mazei. AB - Analysis of genome sequence data from the methanogenic archaeon Methanosarcina mazei Go1 revealed the existence of two open reading frames encoding proton translocating pyrophosphatases (PPases). These open reading frames are linked by a 750-bp intergenic region containing TC-rich stretches and are transcribed in opposite directions. The corresponding polypeptides are referred to as Mvp1 and Mvp2 and consist of 671 and 676 amino acids, respectively. Both enzymes represent extremely hydrophobic, integral membrane proteins with 15 predicted transmembrane segments and an overall amino acid sequence similarity of 50.1%. Multiple sequence alignments revealed that Mvp1 is closely related to eukaryotic PPases, whereas Mvp2 shows highest homologies to bacterial PPases. Northern blot experiments with RNA from methanol-grown cells harvested in the mid-log growth phase indicated that only Mvp2 was produced under these conditions. Analysis of washed membranes showed that Mvp2 had a specific activity of 0.34 U mg (protein)( 1). Proton translocation experiments with inverted membrane vesicles prepared from methanol-grown cells showed that hydrolysis of 1 mol of pyrophosphate was coupled to the translocation of about 1 mol of protons across the cytoplasmic membrane. Appropriate conditions for mvp1 expression could not be determined yet. The pyrophosphatases of M. mazei Go1 represent the first examples of this enzyme class in methanogenic archaea and may be part of their energy-conserving system. PMID- 15803655 TI - Biochemical evidence for the presence of two alpha-glucoside ABC-transport systems in the hyperthermophilic archaeon Pyrococcus furiosus. AB - The hyperthermophilic archaeon Pyrococcus furiosus can utilize different carbohydrates, such as starch, maltose and trehalose. Uptake of alpha-glucosides is mediated by two different, binding protein-dependent, ATP-binding cassette (ABC)-type transport systems. The maltose transporter also transports trehalose, whereas the maltodextrin transport system mediates the uptake of maltotriose and higher malto-oligosaccharides, but not maltose. Both transport systems are induced during growth on their respective substrates. PMID- 15803656 TI - Getting on target: the archaeal signal recognition particle. AB - Protein translocation begins with the efficient targeting of secreted and membrane proteins to complexes embedded within the membrane. In Eukarya and Bacteria, this is achieved through the interaction of the signal recognition particle (SRP) with the nascent polypeptide chain. In Archaea, homologs of eukaryal and bacterial SRP-mediated translocation pathway components have been identified. Biochemical analysis has revealed that although the archaeal system incorporates various facets of the eukaryal and bacterial targeting systems, numerous aspects of the archaeal system are unique to this domain of life. Moreover, it is becoming increasingly clear that elucidation of the archaeal SRP pathway will provide answers to basic questions about protein targeting that cannot be obtained from examination of eukaryal or bacterial models. In this review, recent data regarding the molecular composition, functional behavior and evolutionary significance of the archaeal signal recognition particle pathway are discussed. PMID- 15803658 TI - Loss of genetic accuracy in mutants of the thermoacidophile Sulfolobus acidocaldarius. AB - To investigate how hyperthermophilic archaea can propagate their genomes accurately, we isolated Sulfolobus acidocaldarius mutants exhibiting abnormally high rates of spontaneous mutation. Our isolation strategy involved enrichment for mutator lineages via alternating selections, followed by screening for the production of spontaneous, 5-fluoro-orotate-resistant mutants in micro-colonies. Several candidates were evaluated and found to have high frequencies of pyrE and pyrF mutation and reversion. Neither an increased efficiency of plating of mutants on selective medium, nor the creation of a genetically unstable pyrE allele, could be implicated as the cause of these high frequencies. The strains had elevated frequencies of other mutations, and exhibited certain phenotypic differences among themselves. A large increase in sensitivity to DNA-damaging agents was not observed, however. These properties generally resemble those of bacterial mutator mutants and suggest loss of functions specific to genetic accuracy. PMID- 15803657 TI - Common evolutionary origins of mechanosensitive ion channels in Archaea, Bacteria and cell-walled Eukarya. AB - The ubiquity of mechanosensitive (MS) channels triggered a search for their functional homologs in Archaea. Archaeal MS channels were found to share a common ancestral origin with bacterial MS channels of large and small conductance, and sequence homology with several proteins that most likely function as MS ion channels in prokaryotic and eukaryotic cell-walled organisms. Although bacterial and archaeal MS channels differ in conductive and mechanosensitive properties, they share similar gating mechanisms triggered by mechanical force transmitted via the lipid bilayer. In this review, we suggest that MS channels of Archaea can bridge the evolutionary gap between bacterial and eukaryotic MS channels, and that MS channels of Bacteria, Archaea and cell-walled Eukarya may serve similar physiological functions and may have evolved to protect the fragile cellular membranes in these organisms from excessive dilation and rupture upon osmotic challenge. PMID- 15803659 TI - Characterization of the proteasome from the extremely halophilic archaeon Haloarcula marismortui. AB - A 20S proteasome, comprising two subunits alpha and beta, was purified from the extreme halophilic archaeon Haloarcula marismortui, which grows only in saturated salt conditions. The three-dimensional reconstruction of the H. marismortui proteasome (Hm proteasome), obtained from negatively stained electron micrographs, is virtually identical to the structure of a thermophilic proteasome filtered to the same resolution. The stability of the Hm proteasome was found to be less salt-dependent than that of other halophilic enzymes previously described. The proteolytic activity of the Hm proteasome was investigated using the malate dehydrogenase from H. marismortui (HmMalDH) as a model substrate. The HmMalDH denatures when the salt concentration is decreased below 2 M. Under these conditions, the proteasome efficiently cleaves HmMalDH during its denaturation process, but the fully denatured HmMalDH is poorly degraded. These in vitro experiments show that, at low salt concentrations, the 20S proteasome from halophilic archaea eliminates a misfolded protein. PMID- 15803661 TI - Archaeosomes varying in lipid composition differ in receptor-mediated endocytosis and differentially adjuvant immune responses to entrapped antigen. AB - Archaeosomes prepared from total polar lipids extracted from six archaeal species with divergent lipid compositions had the capacity to deliver antigen for presentation via both MHC class I and class II pathways. Lipid extracts from Halobacterium halobium and from Halococcus morrhuae strains 14039 and 16008 contained archaetidylglycerol methylphosphate and sulfated glycolipids rich in mannose residues, and lacked archaetidylserine, whereas the opposite was found in Methanobrevibacter smithii, Methanosarcina mazei and Methanococcus jannaschii. Annexin V labeling revealed a surface orientation of phosphoserine head groups in M. smithii, M. mazei and M. jannaschii archaeosomes. Uptake of rhodamine-labeled M. smithii or M. jannaschii archaeosomes by murine peritoneal macrophages was inhibited by unlabeled liposomes containing phosphatidylserine, by the sulfhydryl inhibitor N-ethylmaleimide, and by ATP depletion using azide plus fluoride, but not by H. halobium archaeosomes. In contrast, N-ethylmaleimide failed to inhibit uptake of the four other rhodamine-labeled archaeosome types, and azide plus fluoride did not inhibit uptake of H. halobium or H. morrhuae archaeosomes. These results suggest endocytosis of archaeosomes rich in surface-exposed phosphoserine head groups via a phosphatidylserine receptor, and energy-independent surface adsorption of certain other archaeosome composition classes. Lipid composition affected not only the endocytic mechanism, but also served to differentially modulate the activation of dendritic cells. The induction of IL-12 secretion from dendritic cells exposed to H. morrhuae 14039 archaeosomes was striking compared with cells exposed to archaeosomes from 16008. Thus, archaeosome types uniquely modulate antigen delivery and dendritic cell activation. PMID- 15803660 TI - Proteolysis in hyperthermophilic microorganisms. AB - Proteases are found in every cell, where they recognize and break down unneeded or abnormal polypeptides or peptide-based nutrients within or outside the cell. Genome sequence data can be used to compare proteolytic enzyme inventories of different organisms as they relate to physiological needs for protein modification and hydrolysis. In this review, we exploit genome sequence data to compare hyperthermophilic microorganisms from the euryarchaeotal genus Pyrococcus, the crenarchaeote Sulfolobus solfataricus, and the bacterium Thermotoga maritima. An overview of the proteases in these organisms is given based on those proteases that have been characterized and on putative proteases that have been identified from genomic sequences, but have yet to be characterized. The analysis revealed both similarities and differences in the mechanisms utilized for proteolysis by each of these hyperthermophiles and indicated how these mechanisms relate to proteolysis in less thermophilic cells and organisms. PMID- 15803662 TI - Novel archaeal macrocyclic diether core membrane lipids in a methane-derived carbonate crust from a mud volcano in the Sorokin Trough, NE Black Sea. AB - A methane-derived carbonate crust was collected from the recently discovered NIOZ mud volcano in the Sorokin Trough, NE Black Sea during the 11th Training-through Research cruise of the R/V Professor Logachev. Among several specific bacterial and archaeal membrane lipids present in this crust, two novel macrocyclic diphytanyl glycerol diethers, containing one or two cyclopentane rings, were detected. Their structures were tentatively identified based on the interpretation of mass spectra, comparison with previously reported mass spectral data, and a hydrogenation experiment. This macrocyclic type of archaeal core membrane diether lipid has so far been identified only in the deep-sea hydrothermal vent methanogen Methanococcus jannaschii. Here, we provide the first evidence that these macrocyclic diethers can also contain internal cyclopentane rings. The molecular structure of the novel diethers resembles that of dibiphytanyl tetraethers in which biphytane chains, containing one and two pentacyclic rings, also occur. Such tetraethers were abundant in the crust. Compound-specific isotope measurements revealed delta13C values of -104 to 111/1000 for these new archaeal lipids, indicating that they are derived from methanotrophic archaea acting within anaerobic methane-oxidizing consortia, which subsequently induce authigenic carbonate formation. PMID- 15803664 TI - Remarkable sequence signatures in archaeal genomes. AB - Complete archaeal genomes were probed for the presence of long (> or = 25 bp) oligonucleotide repeats (words). We detected the presence of many words distributed in tandem with narrow ranges of periodicity (i.e., spacer length between repeats). Similar words were not identified in genomes of non-archaeal species, namely Escherichia coli, Bacillus subtilis, Haemophilus influenzae, Mycoplasma genitalium and Mycoplasma pneumoniae. BLAST similarity searches against the GenBank nucleotide sequence database revealed that these words were archaeal species-specific, indicating that they are of a signature character. Sequence analysis and genome viewing tools showed these repeats to be restricted to non-coding regions. Thus, archaea appear to possess a non-coding genomic signature that is absent in bacterial species. The identification of a species specific genomic signature would be of great value to archaeal genome mapping, evolutionary studies and analyses of genome complexity. PMID- 15803663 TI - Calcium-induced aggregation of archaeal bipolar tetraether liposomes derived from the thermoacidophilic archaeon Sulfolobus acidocaldarius. AB - Previously, we showed that the proton permeability of small unilamellar vesicles (SUVs) composed of polar lipid fraction E (PLFE) from the thermoacidophilic archaeon Sulfolobus acidocaldarius was remarkably low and insensitive to temperature (Komatsu and Chong 1998). In this study, we used photon correlation spectroscopy to investigate the time dependence of PLFE SUV size as a function of Ca2+ concentration. In the absence of Ca2+, vesicle diameter changed little over 6 months. Addition of Ca2+, however, immediately induced formation of vesicle aggregates with an irregular shape, as revealed by confocal fluorescence microscopy. Aggregation was reversible upon addition of EDTA; however, the reversibility varied with temperature as well as incubation time with Ca2+. Freeze-fracture electron microscopy showed that, after a long period of incubation (2 weeks) with Ca2+, the PLFE vesicles had not just aggregated, but had fused or coalesced. The initial rate of vesicle aggregation varied sigmoidally with Ca2+ concentration. At pH 6.6, the threshold calcium concentration (Cr) for vesicle aggregation at 25 and 40 degrees C was 11 and 17 mM, respectively. At pH 3.0, the Cr at 25 degrees C increased to 25 mM. The temperature dependence of Cr may be attributable to changes in membrane surface potential, which was -22.0 and -13.2 mV at 25 and 40 degrees C, respectively, at pH 6.6, as determined by 2-(p-toluidinyl)naphthalene-6-sulfonic acid fluorescence. The variation in surface potential with temperature is discussed in terms of changes in lipid conformation and membrane organization. PMID- 15803665 TI - Minimal sulfur requirement for growth and sulfur-dependent metabolism of the hyperthermophilic archaeon Staphylothermus marinus. AB - Staphylothermus marinus is an anaerobic hyperthermophilic archaeon that uses peptides as carbon and energy sources. Elemental sulfur (S(o)) is obligately required for its growth and is reduced to H2S. The metabolic functions and mechanisms of S(o) reduction were explored by examining S(o)-dependent growth and activities of key enzymes present in this organism. All three forms of S(o) tested--sublimed S(o), colloidal S(o) and polysulfide--were used by S. marinus, and no other sulfur-containing compounds could replace S(o). Elemental sulfur did not serve as physical support but appeared to function as an electron acceptor. The minimal S(o) concentration required for optimal growth was 0.05% (w/v). At this concentration, there appeared to be a metabolic transition from H2 production to S reduction. Some enzymatic activities related to S(o)-dependent metabolism, including sulfur reductase, hydrogenase, glutamate dehydrogenase and electron transfer activities, were detected in cell-free extracts of S. marinus. These results indicate that S(o) plays an essential role in the heterotrophic metabolism of S. marinus. Reducing equivalents generated by the oxidation of amino acids from peptidolysis may be transferred to sulfur reductase and hydrogenase, which then catalyze the production of H2S and H2, respectively. PMID- 15803667 TI - Specific interactions between aromatic electrons of organic compounds and graphite surfaces as detected by immersion calorimetry. PMID- 15803669 TI - Laser-based patterning of gold nanoparticles into microstructures. PMID- 15803668 TI - Investigation of transversal conductance in semiconductor CdTe nanowires with and without a coaxial silica shell. PMID- 15803671 TI - Electroless nickel-plated UV-embossed microstructured surface with very high aspect ratio channels. PMID- 15803670 TI - Substrates for cell adhesion prepared via active site-directed immobilization of a protein domain. PMID- 15803666 TI - Distribution and phylogenies of enzymes of the Embden-Meyerhof-Parnas pathway from archaea and hyperthermophilic bacteria support a gluconeogenic origin of metabolism. AB - Enzymes of the gluconeogenic/glycolytic pathway (the Embden-Meyerhof-Parnas (EMP) pathway), the reductive tricarboxylic acid cycle, the reductive pentose phosphate cycle and the Entner-Doudoroff pathway are widely distributed and are often considered to be central to the origins of metabolism. In particular, several enzymes of the lower portion of the EMP pathway (the so-called trunk pathway), including triosephosphate isomerase (TPI; EC 5.3.1.1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12/13), phosphoglycerate kinase (PGK; EC 2.7.2.3) and enolase (EC 4.2.1.11), are extremely well conserved and universally distributed among the three domains of life. In this paper, the distribution of enzymes of gluconeogenesis/glycolysis in hyperthermophiles--microorganisms that many believe represent the least evolved organisms on the planet--is reviewed. In addition, the phylogenies of the trunk pathway enzymes (TPIs, GAPDHs, PGKs and enolases) are examined. The enzymes catalyzing each of the six-carbon transformations in the upper portion of the EMP pathway, with the possible exception of aldolase, are all derived from multiple gene sequence families. In contrast, single sequence families can account for the archaeal and hyperthermophilic bacterial enzyme activities of the lower portion of the EMP pathway. The universal distribution of the trunk pathway enzymes, in combination with their phylogenies, supports the notion that the EMP pathway evolved in the direction of gluconeogenesis, i.e., from the bottom up. PMID- 15803672 TI - Self-oscillating nanogel particles. PMID- 15803673 TI - Novel aluminum phosphate-5 crystal morphologies synthesized by microwave heating of a water-in-oil microemulsion. PMID- 15803674 TI - Lanosterol and cholesterol-induced variations in bilayer elasticity probed by 2H NMR relaxation. PMID- 15803675 TI - Modification of black film hydration by infrared irradiation. PMID- 15803676 TI - "Clicking" functionality onto electrode surfaces. PMID- 15803677 TI - Immobilization of gold nanoparticles onto silicon surfaces by Si-C covalent bonds. PMID- 15803678 TI - Structural properties of nonionic cyclodextrin colloids in water. AB - The amphiphilic character in water of a novel class of chemically modified cyclodextrins has been investigated by means of small-angle X-ray scattering and light scattering. The introduction ofhydrophilic oligo(ethylene glycol) onto the secondary side of heptakis[6-alkylthio-6-deoxy-2-oligo(ethylene glycol)]-beta cyclodextrins produces an enhanced water solubility of these molecules. Shape and dimensions of the generated micellar aggregates, analyzed in terms of a suitable core-shell model, remain stable in the wide concentration range explored. The highly associative behavior of these macromolecules is evidenced by the very low value of the critical micelle concentration (cmc), which is about 2 orders of magnitude smaller than the cmc usually obtained for traditional surfactant. Despite the complex geometry of this novel macromolecule, shape and dimensions of generated micellar aggregates can be properly described according to the thermodynamic approaches generally used for amphiphilic molecules and block copolymers. Results show how the modulation of hydrophobic and hydrophilic components sensitively influence the structural features of the generated aggregates thus offering the possibility to control molecular organization in a manner similar to that for traditional colloids. For all the classes of the investigated systems, the small micelles have been found in equilibrium with polydisperse large aggregates of entangled micelles. These novel nonionic colloidal systems combine inclusion and transport properties of host macrocycles, such as cyclodextrin, together with the increased stability of colloidal aggregates, and may be of interest for their potential application as innovative drug delivery systems. PMID- 15803680 TI - Complex coacervation core micelles. Colloidal stability and aggregation mechanism. AB - Complex coacervation core micelles were prepared with various polyelectrolytes and oppositely charged diblock copolymers. The diblock copolymers consist of a charged block and a water-soluble neutral block. Our experimental technique was dynamic light scattering in combination with titrations. At mixing ratios where the excess charge of the polyelectrolyte mixture is approximately zero, micelles may be formed. The colloidal stability of these micelles depends on the block lengths of the diblock copolymers and the molecular weight of the homopolymers. In addition, the chemical nature of the corona blocks and nature of the ionic groups of the polyelectrolytes also influence the stability and aggregation mechanism. A corona block that is three times longer than the core block is a prerequisite for stable micelles. If this ratio is further increased, the molecular weight of the homopolymers as well as the type of the ionic groups starts to play a major role. With very asymmetric block length ratios, no micelles are formed. In addition, if the neutral block is too short, the polymeric mixture forms a macroscopic precipitate. With a constant core block, the aggregation number decreases with increasing corona block length, as is predicted by scaling models for polymeric micelles with a neutral corona. PMID- 15803679 TI - Self-assembly of phosphate fluorosurfactants in carbon dioxide. AB - Anionic phosphodiester surfactants, possessing either two fluorinated chains (F/F) or one hydrocarbon chain and one fluorinated chain (H/F), were synthesized and evaluated for solubility and self-assembly in liquid and supercritical carbon dioxide. Several surfactants, of both F/F and H/F types and having varied counterions, were found to be capable of solubilizing water-in-CO2 (W/C), via the formation of microemulsions, expanding upon the family of phosphate fluorosurfactants already found to stabilize W/C microemulsions. Small-angle neutron scatteringwas used to directly characterize the microemulsion particles at varied temperatures, pressures, and water loadings, revealing behavior consistent with previous results on W/C microemulsions. PMID- 15803681 TI - The adsorption of polymerized rodlike micelles at the solid-liquid interface. AB - Solutions of rodlike polymeric micellar aggregates, formed from the polymerization of cetyltrimethyl-ammonium 4-vinylbenzoate (CTVB), adsorb at the solid-liquid interface. The poly-CTVB aggregates are imaged in situ using soft contact atomic force microscopy. The aggregates form self-organized two dimensional films that show a high degree of order on nanometer to micrometer length scales. Unlike their simple surfactant analogues, the adsorbed layer structures are permanently adsorbed and the structure is resilient to washing with pure solvent. In the case of poly-CTVB, the adsorbed aggregates appear to be rigid cylindrical structures of between 30 and 60 nm in length. At the interface, the center to center spacing of the aligned aggregates is 8+/-1 nm. Images of a second series ofpolymerized aggregates formed by the copolymerization of CTVB with sodium vinyltosylate revealed a change in the aggregate structure to a set of linked spherical aggregates. These polymerized aggregates also spontaneously form a permanent adsorbed layer at the solid-liquid interface. PMID- 15803682 TI - Scattering theory for threadlike micellar solutions. AB - Association-dissociation equilibria and the static scattering function were formulated using precise thermodynamic functions for nonionic surfactant solutions including long, stiff, threadlike micelles. The present theory is applicable for micellar solutions with the surfactant concentration much higher than the critical micelle concentration and containing highly growing threadlike micelles. The scattering function formulated was compared with experimental light scattering data for aqueous solutions of a nonionic surfactant, penta(oxyethylene glycol) n-decyl ether (C12E5), at different surfactant concentrations and also temperatures. PMID- 15803683 TI - Functional polymer colloids with ordered interior. AB - Polymer colloids with internal ordering were synthesized using hydrolytic condensation of octadecyl-dimethyl(3-trimethoxysilylpropyl)ammonium chloride (ODMACl) and a mixture of ODMACl and the trisodium salt of the triacetic acid N (trimethoxysilylpropyl)ethylenediamine (TANED). The structure and morphology of these colloids were studied with small-angle X-ray scattering, transmission electron microscopy, nuclear magnetic resonance, sedimentation in ultracentrifuge; and other methods. When polymer colloids are obtained from a single precursor (ODMACl), their local structure, molecular weight characteristics, and morphology strongly depend on the reaction conditions, while lamellar ordering remains nearly unaffected. Use of a mixture of cationic and anionic silanes (ODMACl and TANED) as precursors in hydrolytic condensation results in novel zwitterionic copolymer colloids with two-dimensional hexagonal packing. Interaction of the ODMACl quaternary ammonium groups with the three carboxy groups of TANED leads to replacement of sodium and chloride ions and formation of gegenions, resulting in a molar ratio ODMACl:TANED = 3:1 (each TANED molecule contains three carboxy groups). Due to their ordered interior, polyODMACl (PODMACl) and PODMACl-TANED colloids can be used as templates for controlled positioning of nanoparticles within these colloids. For example, lamellar ordering controls Pt nanoparticle formation within PODMACl colloids providing Pt nanoparticle alignment within the lamellar structure. Loading of PODMACl-TANED colloids with iron salts followed by pH increase results in the formation of iron oxide nanoparticles located within PODMACl-TANED cylinders. PMID- 15803684 TI - Multitechnique characterization of fatty acid-modified microgels. AB - The structure of fatty acid-modified hydrogel objects (microgels) created within microfluidic devices for controlled-release or sensory applications was characterized by various imaging and spectroscopic methods. Imaging with scanning electron microscopy revealed that the surface was rough and irregular on the micrometer scale. Examination of planar model systems analogous to the modified microgels with X-ray photoelectron spectroscopy and secondary ion mass spectroscopy showed that a fatty acid coating formed when the reaction conditions were conducive to covalent-bond formation. Visualization of the selectively stained lipophilic coating with laser scanning confocal microscopy while the microgel was positioned within a microfluidic channel demonstrated that the coating was confined to the microgel's periphery. Finally, using transmission electron microscopy, the thickness of the region functionalized with fatty acids was determined to be around 9 microm on samples very similar to those integrated into microfluidic devices. By using transmission electron microscopy to monitor the thickness of the fatty acid coating produced with different reaction conditions, it may be possible to customize these hybrid materials for specific sensory or controlled-release applications. PMID- 15803685 TI - Light-sensitive microemulsions. AB - A photodestructible surfactant, sodium 4-hexylphenylazosulfonate (C6-PAS), has been introduced to AOT-stabilized water-in-heptane microemulsions. Proton NMR spectra show that C6-PAS undergoes UV-induced decomposition, to yield a mixture of 4-hexylphenol and hexylbenzene. The photostationary state was determined by 1H NMR, indicating that nearly 90% of the initial photosurfactant had been destroyed, yielding non-surface-active hexylbenzene as the main product. This phototriggered breakdown gives rise to changes in adsorption and aggregation properties of C6-PAS, representing a novel route to induce microemulsion destabilization. When a series of microemulsions containing different amounts of C6-PAS were exposed to UV light, part of the dispersed water phase-separated. Small-angle neutron scattering (SANS) was used to follow the resulting UV-induced shrinkage of the water nanodroplets: a maximum volume decrease was found to be in the order of 60-70%. Kinetic SANS studies were also carried out in order to follow the changes in aggregation as a function of UV irradiation time. Multicontrast SANS experiments gave further insight; for example, it was demonstrated that the shell thickness remained constant. This study represents the first example of light-induced microemulsion destabilization. PMID- 15803686 TI - Rheo-optical properties of silicone oil emulsions in the presence of polymer emulsifiers. AB - Oil in water emulsions prepared by dispersion of silicone oils into an aqueous solution of hydroxylpropyl methyl cellulose (HPMC) or poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) block copolymers were characterized by measurements of steady-state shear viscosities, dynamic moduli, and stress-strain sweep curves coupled with optical microscopic observation. The emulsions prepared with HPMC showed solidlike viscoelastic responses and a clear yield stress, whereas the emulsions emulsified by PEO-PPO-PEO block copolymers indicated liquidlike viscoelastic behavior. The difference should be attributed to stronger protective colloidal effects, that is, the former emulsifiers form a more viscoelastic polymer layer adsorbed on the oil droplets than the latter ones. Moreover, the simultaneous optical microscopic observation showed that the emulsions stabilized by HPMC do not flow below the yield stress and beyond the yield stress the movements of oil droplets occur first. PMID- 15803687 TI - Silica particle-stabilized emulsions of silicone oil and water: aspects of emulsification. AB - A study of the emulsification of silicone oil and water in the presence of partially hydrophobic, monodisperse silica nanoparticles is described. Emulsification involves the fragmentation of bulk liquids and the resulting large drops and the coalescence of some of those drops. The influence of particle concentration, oil/water ratio, and emulsification time on the relative extents of fragmentation and coalescence during the formation of emulsions, prepared using either batch or continuous methods, has been investigated. For batch emulsions, the average drop diameter decreases with increasing particle concentration as the extent of limited coalescence is reduced. Increasing the oil volume fraction in the emulsion at fixed aqueous particle concentration results in an increase in the average drop diameter together with a dramatic lowering of the uniformity of the drop size distribution as coalescence becomes increasingly significant until catastrophic phase inversion occurs. For low oil volume fractions (phi(o)), fragmentation dominates during emulsification since the mean drop size decreases with emulsification time. For higher phi(o) close to conditions of phase inversion, coalescence becomes more prevalent and the drop size increases with time with stable multiple emulsions forming as a result. PMID- 15803688 TI - Surfactant/Nonionic polymer interaction. A NMR diffusometry and NMR electrophoretic investigation. AB - The interaction between the nonionic polymer poly(ethylene oxide) (PEO) of molecular weight 20,000 and surfactants of various types [sodium dodecyl sulfate (SDS), dodecyl trimethylammonium bromide, octyl beta-D-glucoside, and potassium laurate] has been investigated in an aqueous solution at 25 degrees C by 1H NMR pulsed-gradient spin-echo self-diffusion techniques. The SDS/PEO study was further complemented by component-resolved 1H NMR-based studies of the electrophoretic mobility of PEO and the alkyl part of SDS under the same measurement conditions. Through such combined studies, a much more complete picture of the binding and aggregation processes becomes accessible. PMID- 15803689 TI - Multilamellar liposomes formed by phosphatidyl nucleosides: an NMR-HR-MAS characterization. AB - We present an NMR investigation of multilamellar vesicles (MLVs) obtained from phosphatidyl nucleosides, 5'-(1-palmitoyl-2-oleoyl-sn-glycero(3)phospho)cytidine (1), 5'-(1-palmitoyl-2-oleoyl-sn-glycero(3)phospho)inosine (2), and their mixtures. Because of the lower stability of liposomes obtained from 2, studies have been preferentially performed in this case with mixed liposomes 2/POPC (4:1). The investigation is conducted mostly via the HR-MAS technique and the general observation is that the resolution achieved in this way is superior to that obtained in the past with small unilamellar vesicles (SUVs). A full assignment is now possible, which includes the spectral region of the ribose ring and part of the glycerol moiety. Also in the case of MLVs, both for 1 and 2, a stacking between the aromatic bases of the same liposome layer seems to be ruled out, although in both cases the nucleobases appear to be exposed to the aqueous phase. The splitting of both aromatic H-5cyt and H-6cyt is ascribed to the presence of two aggregate populations that may correspond to the two syn and anti conformations observed for cytidine monophosphate in aqueous solution. On the basis of NOESY cross-peaks, it is not always possible to discriminate between inter- and intramolecular interactions; however, the distances found for 1 appear to be compatible with the intramolecular contacts in the anti conformation of the cytidine and also with intermolecular interactions between neighboring molecules of 1. We also find that the glycerol moiety does not seem to interact with the cytidine; however, part of the ribose ring seems to be close to the glycerol moiety. More generally, the interaction of one base with the sugar moiety of a neighboring base, previously observed for SUVs, still appears to be true for MLVs. Studies have been performed also for mixed liposomes obtained from the mixture of 1 and 2, where it is observed that the HR-MAS spectra of the corresponding MLVs are not simply the sum of the spectra of the two isolated components. In particular, there is the presence of a NOESY cross-peak between the aromatic protons H-6cyt and H-2ino, and this permits us to rule out large patchwork domains containing only one nucleoside components in the mixed liposomes. Finally, a study is performed on the time evolution of the system obtained by mixing the previously prepared liposomes of 1 and 2. No interaction is obtained in this case, i.e., the spectra are constitutive, which is consistent with the general picture of liposomes as kinetic traps that are not fusing with each other. PMID- 15803691 TI - Interaction between a partially fluorinated alkyl sulfate and gelatin in aqueous solution. AB - The interaction of a partially fluorinated alkyl sulfate, sodium 1H,1H,2H,2H perfluorooctyl sulfate (C6F13CH2CH2OSO3Na), with the polyampholyte gelatin has been examined in aqueous solution using surface tension and small-angle neutron scattering (SANS). The 19F chemical shift of each fluorine environment in the surfactant is unaltered by the addition of gelatin, indicating that there is no contact between the gelatin and the fluorocarbon core of the micelle. The chemical shift of the two methylene groups closest to the headgroup is altered when gelatin is present, disclosing the location of the polymer. The critical micelle concentration (cmc) of the surfactant, cmc = 17+/-1 mM, corresponds to an effective alkyl chain (CnH2n+1) length of n = 11. In the presence of gelatin, the cmc is substantially reduced as expected, cmc(1) = 4+/-1 mM, which is also consistent with an effective alkyl chain length of n = 11. In the presence of the fluorosurfactant, the monotonic decay of the SANS from the gelatin-only system is replaced by a substantial peak at an intermediate Q value mirroring the micellar interaction. At low ionic strengths, the gelatin/micelle complex can be described by an ellipsoid. At higher ionic strengths, the electrostatic interaction between the micelles is screened and the peak in the gelatin scattering disappears. The correlation length describing the network structure decreases with increasing SDS concentration as the bound micelles promote a collapse of the network. PMID- 15803690 TI - H-aggregation of azobenzene-substituted amphiphiles in vesicular membranes. AB - Photochemical switching has been studied of double-tailed phosphate amphiphiles containing azobenzene units in both tails in aqueous vesicular dispersions and in mixed vesicular systems with 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). Since the ease of switching depends on the strength of the bilayer packing, particular emphasis has been placed on the occurrence of H-aggregation in the hydrophobic core of the vesicles. UV-vis spectrometry was employed to monitor H aggregation and showed how this process depends on the ionic strength and on the mode of preparation of the vesicles. Two types of H-aggregates were observed in mixed DOPC vesicles with 5 mol % of azobenzene phosphate: one with lambda(max) at around 300 nm and one with lambda(max) at 305-320 nm. Those with lambda(max) at 300 nm could not be trans-cis photoisomerized, whereas those with lambda(max) at 305-320 nm are more loosely packed and can be photochemically switched. The permeability of the vesicular bilayers, as probed with leakage experiments using calcein as a fluorescent probe, was examined as another measure for the strength of bilayer packing. Leakage occurred only for DOPC vesicles containing more than 20 mol % of azobenzenephosphate, irradiated with UV light to induce trans-cis photoisomerization. We contend that detailed information on bilayer packing will be of crucial importance for fine-tuning the lateral pressure in vesicular membranes with the ultimate aim to steer the opening and closing of mechanosensitive protein channels of large conductance. PMID- 15803692 TI - Phase behavior of 1-alkylpyridinium octane-1-sulfonates. effect of the 1 alkylpyridinium counterion size. AB - The temperature-versus-composition phase diagrams of eight different 1 alkylpyridinium octane-1-sulfonates (APOSs) in water were studied by 1H NMR, 2H NMR, pulsed gradient spin-echo NMR, small-angle X-ray diffraction, differential scanning calorimetry, surface tension and conductivity measurements, and polarizing microscopy. The number of carbons (n(c)) in the hydrocarbon chain of the pyridinium counterions was varied from n(c) = 1 to n(c) = 8 to study how the phase behavior of the APOS/2H2O systems was affected by a change in the chain length of the counterion. The sodium octane-1-sulfonate (NaOS)/water system was used as a reference. This system formed an isotropic micellar solution (L1) phase and a normal hexagonal (H(I)) phase. All APOSs were readily soluble in water and formed L1 phases. The surface tension above the critical micelle concentration for n(c) between 1 and 3 was higher than that for NaOS, and it decreased steadily for the different APOSs with increasing chain length. The area per molecule at the air/solution interfaces was rather constant at 68 A2 for n(c) between 1 and 7. For 1-octylpyridinium octane-1-sulfonate (OPOS), it was about 5 A2 smaller, which was just outside the estimated error. However, the smallest area was obtained for NaOS. At higher surfactant concentrations, liquid crystalline phases formed. Both cubic and H(I) phases were found for n(c) = 1 and 2, while for n(c) between 3 and 5 only an H(I) phase was observed. H(I) and lamellar liquid crystalline (Lalpha) phases formed for n(c) = 6 and 7. The only liquid crystalline phase found in the OPOS system was a Lalpha phase. The NaOS H(I) phase was the only liquid crystalline phase that showed a linear relation between the 2H2O NMR quadrupolar splitting (deltaW) and Xsurf/X(W), where Xsurf and X(W) are the mole fractions of surfactant and water. The OPOS lamellae were found to be much thinner than expected, indicating a defect lamellar structure. This was further supported by the behavior of the quadrupolar splitting ofdeuterated OPOS. The anomalous behaviors of the 2H2O NMR quadrupolar splitting observed in the Lalpha phases of 1-heptylpyridinium octane-1-sulfonate and OPOS were interpreted in terms of changes in the population of the water molecules residing in different sites combined with a continuous rearrangement of the lamellae surface with the possible development of holes. The appearances of the phase diagrams were discussed in terms of surfactant molecular geometry and the packing of the amphiphiles in the aggregates formed. PMID- 15803693 TI - Sorption rate and thermal barriers in a gas-zeolite system: investigation of n hexane sorption in MFI-type zeolite. AB - The nonequilibrium gravimetric sorption method was used to determine diffusion coefficient values for n-hexane in MFI-type materials. Improvements in the measurement device and experimental conditions resulted in high values of the corrected diffusion coefficient, which are comparable to the literature data obtained by the methods of pulsed field gradient nuclear magnetic resonance (PFG NMR) and frequency response (FR). The results indicate that thermal effects of sorption affect practically neither the rate of the sorption nor the diffusion coefficient. PMID- 15803694 TI - Covalent attachment of a nickel nitrilotriacetic acid group to a germanium attenuated total reflectance element. AB - The surface of a germanium internal reflectance element (IRE) was modified to bind 6X-histidine (his)-tagged biomolecules. The step-by-step surface modification was monitored via single-pass attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FT-IR). Initially an adlayer of 7 octenyltrimethoxysilane (7-OTMS) was formed on the Ge crystal through the surface hydroxyl groups, which were produced via ozonolysis of the Ge surface. The vinyl moiety of 7-OTMS was oxidized to a carboxylic acid, which was activated by 1,1' carbonydiimidazole (CDI) to produce a labile imidazole. The labile imidazole that resulted from the CDI coupling was then displaced by the primary amine of nitrilotriacetic acid (NTA). Nickel sulfate was added to the system, and it coordinated with the three carbonyl groups and the nitrogen on NTA, thus leaving the ability of Ni to coordinate with two adjacent histidine residues. Binding of his-tagged biotin to nickel nitrilotriacetic acid (Ni-NTA) was observed by ATR-FT IR spectroscopy. The surface modification method presented in this paper had minimal nonspecific binding, the Ni-NTA surface was reusable if stored properly, and complete removal of the organic surface was achievable. PMID- 15803695 TI - Novel dual-bed reactors: utilization of hydrogen spillover in reactor design. AB - Hydrogen spillover over macroscopic distances was demonstrated and exploited in the design of two novel catalytic reactors for 1-butene isomerization. A dual-bed reactor containing separate zones of noble metal and bimetallic catalysts yielded activities up to 2.7 times greater than that of the noble metal alone. The noble metal catalyst contained palladium supported on graphitic carbon. The bimetallic catalyst contained a base metal, either iron or cobalt, and a lanthanide metal, either cerium or praseodymium, also supported on graphitic carbon. The bimetallic catalysts by themselves had no measurable activity at the current experimental conditions. Results from a dual-bed, dual-feed reactor using the same catalysts showed dramatic activity increases relative to controls. In this reactor, the hydrocarbon never contacted the noble metal catalyst, yet substantial hydrocarbon conversion was measured. No hydrocarbon conversion was detected when blank support replaced the bimetallic catalyst or when no material at all was placed above the noble metal catalyst. In both reactors, the activity increase was attributed to hydrogen spillover. That is, molecular hydrogen adsorbed and dissociated on the noble metal catalyst. The adsorbed atomic hydrogen was then transported via surface diffusion to the bimetallic catalyst, activating those sites. The results also demonstrated that a catalytic reaction may occur at distinctly different reactive sites and that catalysts may be selected to promote specific steps within the reaction. PMID- 15803696 TI - Nanoporous carbon films for gas microsensors. AB - We study nanoporous carbon (NPC) as an adsorbent coating on surface acoustic wave (SAW) chemical microsensors for a wide range of analyte gases. By use of pulsed laser deposition in a controlled inert gas ambient, NPC grows at room temperature with negligible residual stress and, hence, can coat most surfaces to any desired thickness. Acetone adsorption isotherms for NPC-coated SAW devices with mass density ranging from 0.18 to 1.08 g/cm3 indicate that the device frequency response relates to NPC density. Data analysis suggests the possibility of detecting acetone below parts-per-billion concentrations. We find NPC to be highly sensitive to a variety of other volatile organic and toxic industrial compounds. Transmission electron microscopy reveals that lower-density NPC has both larger and greater numbers of nanopores than higher-density NPC and that decreasing NPC density also increases the interplanar spacing between graphene sheet fragments within the ultrathin carbon wall structures. These physical differences effectively increase the available surface area for analyte gas adsorption with decreasing NPC density, with only the structural integrity of the internal NPC wall structures a limiting factor in determining the lowest useful density NPC coating. PMID- 15803697 TI - Diffusion-limited thiol adsorption on the gold(111) surface. AB - An optical second harmonic generation measurement of the kinetics of self assembly of a monolayer of thiols on the Au(111) surface reveals a marked dependence of the adsorption rate upon the solution flow rate. The nature of this dependence indicates that at low concentration and low flow rate the monolayer growth is limited by the existence of a Nernst diffusion layer, not by surface reaction rate kinetics. PMID- 15803698 TI - Decomposition processes of an organic monolayer formed on Si(111) via a silicon carbon bond induced by exposure to UV irradiation or ozone. AB - The decomposition processes of an organic monolayer, which was formed on Si(111) via a Si-C covalent bond, induced by exposure to UV light irradiation or ozone, were investigated using attenuated total reflectance Fourier transform infrared spectroscopy. Exposure to both ozone and UV light resulted in a reduction of the intensities of the IR peaks corresponding to CH stretching vibration and bending scissors and the appearance of peaks corresponding to CO stretching and COH in plane bending. The latter peaks initially increased, reached a maximum, and then decreased, indicating that the monolayer was decomposed through the formation of intermediates such as aldehyde and carboxylic acid. The monolayer was also decomposed by exposure only to UV light or ozone but more slowly as the time dependencies of the CH peaks showed. While the peaks corresponding to the CO stretching and the COH in-plane bending behaved similarly under the condition of exposure to ozone, they were not observed during decomposition induced by UV irradiation. These results show that, while the monolayer was decomposed through the formation of oxidized intermediates such as aldehyde and carboxylic acid under the condition of exposure to ozone, the decomposition of the monolayer under the condition of UV irradiation proceeded via cleavage of Si-C bonds by photogenerated electrons or holes without such oxidized intermediates. An increase of gauche defects as the decomposition proceeded was demonstrated by sum frequency generation spectroscopy. PMID- 15803699 TI - Contact line dynamics in drop coalescence and spreading. AB - The dynamics of coalescence of two water sessile drops is investigated and compared with the spreading dynamics of a single drop in partially wetting regime. The composite drop formed due to coalescence relaxes exponentially toward equilibrium with a typical relaxation time that decreases with contact angle. The relaxation time can reach a few tenths of seconds and depends also on the drop size, initial conditions, and surface properties (contact angle, roughness). The relaxation dynamics is larger by 5 to 6 orders of magnitude than the bulk hydrodynamics predicts, due to the high dissipation in the contact line vicinity. The coalescence is initiated at a contact of the drops growing in a condensation chamber or by depositing a small drop at the top of neighboring drops with a syringe, a method also used for the studies of the spreading. The dynamics is systematically faster by an order of magnitude when comparing the syringe deposition with condensation. We explain this faster dynamics by the influence of the unavoidable drop oscillations observed with fast camera filming. Right after the syringe deposition, the drop is vigorously excited by deformation modes, favoring the contact line motion. This excitation is also observed in spreading experiments while it is absent during the condensation-induced coalescence. PMID- 15803700 TI - Irradiation sensitivity of self-assembled monolayers with an introduced "weak link". AB - The irradiation sensitivity of n-alkanethiolate SAMs can be enhanced by the incorporation of an irradiation-sensitive functionality as a predetermined breaking point into the alkyl chain. Following this general strategy, several different sulfur-derived functional entities with a variable position in the alkyl chain were tested. In some cases, the incorporation of these entities resulted in a noticeable enhancement of the irradiation-induced desorption (by about 30%), even though the chains were not exclusively cut at the position of the incorporated group, but the scission events related to the "weak links" were just added to those in the alkyl matrix. The position of the incorporated group along the alkyl chain was found to be most essential for its performance as a weak link; the enhancement of irradiation-induced desorption was observed only when the groups were placed close to the SAM-ambient interface. PMID- 15803701 TI - Preparation and heck reaction of multidentate carbosilane films derived from focally functionalized and allyl-terminated dendrons on hydrogen-terminated silicon(111) surfaces. AB - Multidentate carbosilane films were prepared by thermally induced hydrosilylation of allyl-terminated carbosilane dendrons of generations 0, 1, and 2 (G0-G2) on hydrogen-terminated silicon(111) surfaces. The dendron molecules contain three (G0), nine (G1), and twenty-seven (G2) allyl groups at the periphery, and a bromophenyl functional group at the focal point. The dendron films were characterized by contact-angle goniometry, ellipsometry, Fourier transform infrared spectroscopy in the attenuated total reflection mode, and X-ray photoelectron spectroscopy (XPS). Upon hydroboration of the remaining allyl groups in the films, the percentage of the introduced boron atoms in the films were measured by XPS. The results indicate the presence of roughly 20%, 27%, and 46% of unreacted allyl groups in the G0, G1, and G2 films, respectively. The mechanistic aspects of the chemisorption of these dendron molecules on H-Si(111) surfaces are discussed. XPS studies indicate that seven G0 molecules cover approximately the same area on the substrate as three G1 molecules and one G2 molecule. After treatment of the G0, G1, and G2 films with 4-fluorostyrene under the Heck reaction conditions, the XPS studies indicate that about 84%, 71%, and 55% of the Br atoms were consumed, yielding the replacement of ca. 58-70% of the reacted Br atoms by the fluorostyryl groups. The remaining bromophenyl groups were inactive toward the Heck reaction, probably due to their disfavorable position/orientation in the films. PMID- 15803702 TI - Electrohydrodynamic effect on phase separation morphology in polymer blend films. AB - We have investigated the effect of electrohydrodynamic (EHD) convection on the domain structure in a polystyrene (PS)/polyvinyl acetate (PVA) blend film to demonstrate the feasibility of using the EHD effect as a means of mixing and morphology control in a polymer blend film prepared by solvent evaporation. Here, polymers-toluene solutions were spread on a glass substrate with patterned electrodes to apply a dc electric field, and well-defined structures of EHD convection were formed in the polymer solutions. As a result, regular patterns were formed in the PS/PVA polymer blend film in which PVA-rich domains were confined within each unit of patterned electrodes, i.e., between positive and negative electrodes, at an appropriate electric voltage. In addition, it was demonstrated that such novel morphology is not due to the wetting/dewetting effect of polymer components to the Pt electrodes deposited on the glass substrate, by experiments with a SiO2-covered substrate. PMID- 15803703 TI - Effect of high-viscosity interphases on drainage between hydrophilic surfaces. AB - Drainage of water from the region between an advancing probe tip and a flat sample is reconsidered under the assumption that the tip and sample surfaces are both coated by a thin water "interphase" (of width approximately a few nanometers) whose viscosity is much higher than that of the bulk liquid. A formula derived by solving the Navier-Stokes equations allows one to extract an interphase viscosity of approximately 59 kPa x s (or approximately 6.6 x 10(7) times the viscosity of bulk water at 25 degrees C) from interfacial force microscope measurements with both tip and sample functionalized hydrophilic by OH terminated tri(ethylene glycol) undecylthiol, self-assambled monolayers. PMID- 15803704 TI - Adlayers of C60-C60 and C60-C70 fullerene dimers formed on au(111) in benzene solutions studied by STM and LEED. AB - Scanning tunneling microscopy (STM) and low-energy electron diffraction were used to reveal the structures of ordered adlayers of [2+2]-type C60-C60 fullerene dimer (C120) and C60-C70 cross-dimer (C130) formed on Au(111) by immersingit in abenzene solution containing C120 or C130 molecules. High-resolution STM images clearly showed the packing arrangements and the electronic structures of C120 and C130 on the Au(111) surface in ultrahigh vacuum. The (2 square root3 x 4square root3)R30 degrees, (2square root3 x 5square root3)R30 degrees, and (7 x 7) structures were found for the C120 adlayer on the Au(111) surface, whereas C130 molecules were closely packed on the surface. Each C60 or C70 monomer cage was discerned in the STM image of a C130 molecule. PMID- 15803705 TI - Spreading and recoil of a surfactant-containing water drop on glass-supported alcohol films. AB - In this paper we report the experimental observation of spreading and recoil of surfactant-containing water drops on various alcohol films supported on glass slides. The time evolution of spreading and recoil behavior was recorded by placing a web camera above the drop. We observed that the drop spread the fastest on CH3OH, followed by C2H5OH, and the slowest on i-PrOH. On the other hand, the recoil behavior was just the opposite. The drop recoiled the slowest on CH3OH and fastest on i-PrOH, while it recoiled in an intermediate time on C2H5OH. In addition, concentration of surfactant in the drop played a prominent role in the spreading and recoil time of the drop, with higher surfactant concentration making the drop spread and recoil faster. The time evolution of spreading velocity of the drop on different alcohol films at various surfactant concentrations occurred with a Gaussian distribution and the peak velocity was reached earliest on CH3OH followed by C2H5OH, while on i-PrOH it took the longest time. The recoil behavior was similar. The variation of velocity as a function of radius exhibited oscillatory behavior, indicating the existence of an interfacial phenomenon. We also report the observation that spreading of the drop occurred without observable fingering instability. Further, we observed by Fourier transform infrared (FTIR) spectroscopic measurements that the drop had mixed with the alcohol films as it spread. Miscibility of the alcohol in the film with the drop, alcohol evaporation cooling-induced temperature gradient, and Marangoni effect probably play important roles in the spreading and recoil behavior of the drop. PMID- 15803706 TI - Dynamic in situ characterization of organic monolayer formation via a Novel substrate-mediated mechanism. AB - Ultrahigh vacuum scanning tunneling microscopy data investigating octylsilane (C8H17SiH3) monolayer pattern formation on Au(111) are presented. The irregular monolayer pattern exhibits a 60 A length scale. Formation of the octylsilane monolayer relaxes the Au(111) 23 x square root3 surface reconstruction and ejects surface Au atoms. Au adatom diffusion epitaxially extends the Au(111) crystal lattice via step edge growth and island formation. The chemisorbed monolayer covers the entire Au surface at saturation exposure. Theoretical and experimental data suggest the presence of two octylsilane molecular adsorption phases: an atop site yielding a pentacoordinate Si atom and a surface vacancy site yielding a tetracoordinate Si atom. Theoretical simulations investigating two-phase monolayer self-assembly dynamics on a solid surface suggest pattern formation results from strain-induced spinodal decomposition of the two adsorption phases. Collectively, the theoretical and experimental data indicate octylsilane monolayer pattern formation is a result of interfacial Au-Si interactions and the alkyl chains play a negligible role in the monolayer pattern formation mechanism. PMID- 15803707 TI - Surface and solution behavior of the mixed dialkyl chain cationic and nonionic surfactants. AB - The surface and solution behavior of the mixed dialkyl chain cationic and nonionic surfactant mixture of dihexadecyldimethylammonium bromide, DHDAB, and hexaethylene monododecyl ether, C12E6, has been investigated, using primarily the scattering techniques of small-angle neutron scattering and neutron reflectivity. Within the time scale of the measurements, the adsorption of the pure component C12E6 at the air-solution interface shows no time dependence. In contrast, the adsorption of the DHDAB/C12E6 mixture and pure DHDAB has a pronounced time dependence. The characteristic time for adsorption varies with surfactant concentration, composition, and temperature. It is approximately 2-3 h for the DHDAB/C12E6 mixture, dependent upon concentration and composition, and approximately 50 min for DHDAB. At the air-solution interface, the equilibrium composition of the adsorbed layer shows a marked departure from ideal mixing, which is dependent upon both the solution concentration and the concentration of added electrolyte. In contrast, the composition of the aggregates in the bulk solution that are in equilibrium with the surface is close to ideal mixing, as expected for solution concentrations well in excess of the critical micellar concentration. The structure of the mixed adsorbed layer has been measured and compared with the structure of the equivalent pure surfactant monolayer, and no substantial changes in structure or conformation are observed. The extreme departure from ideal mixing in the adsorption behavior of the DHDAB/C12E6 mixture is discussed in the context of the structure of the adsorbed layer, changes in the underlying solution structures, and the failure of regular solution theory to predict such behavior. PMID- 15803708 TI - Interaction of water with self-assembled monolayers of alkylsilanes on mica. AB - The interaction of water with self-assembled alkylsilane monolayers on mica substrates has been studied using an atomic force microscope operated in contact, noncontact, and electrical polarization modes. Complete monolayer films were found to be effective in blocking water adsorption. On partially covered surfaces water was found to produce large changes in the conductivity and surface potential of the exposed mica regions. It was also found that water could penetrate films near defects and at island edges. PMID- 15803709 TI - Direct visualization of mesh structures at solid/solution interfaces by atomic force microscopy. AB - The formation of adsorbed surfactant layers consisting of a mesh or network of branched cylindrical aggregates on muscovite mica by several surfactant systems is described. The curvature of the adsorbed aggregates is varied by a variety of mechanisms that all generate morphologies between adsorbed cylinders and bilayers, and the resulting lateral structure is imaged by "soft contact" atomic force microscopy. We compare the direct images and Fourier transforms of the adsorbed layer structures, and relate them to those formed in bulk solution. PMID- 15803710 TI - Self-assembled metal colloid films: two approaches for preparing new SERS active substrates. AB - In this paper, we propose two new approaches for preparing active substrates for surface-enhanced Raman scattering (SERS). In the first approach (method 1), one transfers AgI nanoparticles capped by negatively charged mercaptoacetic acid from a AgI colloid solution onto a quartz slide and then deoxidizes AgI to Ag nanoparticles on the substrate. The second approach (method 2) deoxidizes AgI to Ag nanoparticles in a colloid solution and then transfers the Ag nanoparticles capped by negatively charged mercaptoacetic acid onto a quartz slide. By transfer of the AgI/Ag nanoparticles from the colloid solutions to the solid substrates, the problem of instability of the colloid solutions can largely be overcome. The films thus prepared by both approaches retain the merits of metal colloid solutions while they discharge their shortcomings. Accordingly, the obtained Ag particle films are very suitable as SERS active substrates. SERS active substrates with different coverages can be formed in a layer-by-layer electrostatic assembly by exposing positively charged surfaces to the colloid solutions containing oppositely charged AgI/Ag nanoparticles. The SERS active substrates fabricated by the two novel methods have been characterized by means of atomic force microscopy (AFM) and ultraviolet-visible (UV-vis) spectroscopy. The results of AFM and UV-vis spectroscopy show that the Ag nanoparticles grow with the increase in the number of coverage and that most of them remain isolated even at high coverages. Consequently, the surface optical properties are dominated by the absorption due to the isolated Ag nanoparticles. The relationship between SERS intensity and surface morphology of the new active substrates has been investigated for Rhodamine 6G (R6G) adsorbed on them. It has been found that the SERS enhancement depends on the size and aggregation of the Ag particles on the substrates. Especially, we can obtain a stronger SERS signal from the substrate prepared by method 1, implying that for the metal nanoparticles capped with stabilizer molecules such as mercaptoacetic acid, the in situ deoxidization in the film is of great use in preparing SERS active substrates. Furthermore, we have found that the addition of Cl- into the AgI colloid solution changes the surface morphology of the SERS active substrates and favors stronger SERS enhancement. PMID- 15803711 TI - Gibbs monolayers at the air/water interface: surface partitioning and lateral mobility of an electrochemically active surfactant. AB - Monolayer films of a water-soluble surfactant, 4-octaneamido-2,2,6,6-tetramethyl 1-piperidinyloxy (C8-TEMPO) were investigated at the air/water interface. An electrochemical, horizontal touch method was developed to measure the equilibrium surface concentrations (gamma) of C8TEMPO. The dependence of gamma on the solution concentration followed a Langmuir isotherm and yielded the partition constant K = (2.3+/-0.2) x 10(4) M(-1). These results were verified by surface tension measurements and Brewster angle microscopy. Within experimental error, the same K values were obtained. The lateral diffusion constants vs surface concentration of this molecule were measured by 2D voltammetry. In these experiments, the component of the oxidation current due to C8TEMPO in the bulk of the solution was subtracted from the total measured current to obtain the component due to the lateral surface diffusion. In the ange of mean molecular areas from 120 to 400 A2/molecule, the lateral diffusion constant of C8TEMPO increased from 1.0 x 10(-6) to 1.0 x 10(-5) cm2/s. The latter value is about 2.5 times larger than the C8TEMPO diffusion constant in bulk water. Comparison of the lateral mobilities of C8TEMPO and two longer alkane chain, water-insoluble homologues, C14TEMPO and C18TEMPO, showed no statistically significant differences. PMID- 15803712 TI - Isostructural self-assembled monolayers. 2. Methyl 1-(3-mercaptopropyl) oligo(ethylene oxide)s. AB - The structural order and ordering conditions of the self-assembled monolayers (SAMs) of HSCH2CH2CH2O(EO)xCH3, where EO = CH2CH2O and x = 3-9, on polycrystalline gold (Au) were determined by reflection-absorption infrared spectroscopy (RAIRS), spectroscopic ellipsometry (SE), and electrochemical impedance spectroscopy. For x = 5-7, RAIRS and SE data show that the oligo(ethylene oxide) [OEO] segments adopt the near single phase, 7/2 helical conformation of the folded-chain crystal polymorph of crystalline poly(ethylene oxide), oriented normal to the substrate. These SAMs exhibit OEO segment structure and orientation identical to that found in a previous isostructural series [HS(CH2CH2O)6-8C18H37 SAMs. Vanderah, D. J., et al. Langmuir 2003, 19, 3752] and are anisotropic films for surface science metrology where structure is constant and thickness increases in 0.30 nm increments. In addition, this is the first example of OEO SAMs to attain this highly ordered, helical conformation where the (EO)x segment is separated from the Au-sulfur headgroup by a polymethylene chain. For x = 4, 8, and 9, the SAMs are largely helical but show evidence of nonhelical conformations and establish the upper and lower limits of the isostructural set. For x = 3, the SAMs are largely disordered containing some all-trans conformation. SAM order as a function of immersion time from 100% water and 95% ethanol indicates that the HSCH2CH2CH2O(EO)5-7CH3 SAMs order faster and under a wider range of conditions than omega-alkyl 1-thiaolio(ethylene oxide) [HS(EO)xCH3] SAMs, reported earlier (Vanderah, D. J., et al. Langmuir 2002, 18, 4674 and Vanderah, D. J., et al. Langmuir 2003, 19, 2612). PMID- 15803713 TI - Adhesion and wetting in an aqueous environment: theoretical assessment of sensitivity to the solid surface energy. AB - The sensitivity of contact angles in an aqueous environment to the surface energy of the solid is discussed. It is demonstrated that this sensitivity is much higher in an aqueous than in a vapor environment. The Girifalco-Good equation, in combination with the Owens-Wendt equation, is used for the approximate demonstrations. It is shown that the transition from complete wetting to complete dewetting by the aqueous phase in a solid-liquid-liquid system occurs over a much narrower range of the surface energy of the solid than in a solid-liquid-vapor system. It is also demonstrated that the contact angle may be extremely sensitive to small variations in the relationship between surface tensions and the corresponding interfacial tension. PMID- 15803714 TI - Morphological investigation of chemically treated poly(ethylene terephthalate) based activated carbons. AB - Complementary techniques, including low-temperature nitrogen adsorption and small angle X-ray scattering (SAXS), are applied to detect the effects of surface functionalization on the morphology of activated carbon derived from poly(ethylene terephthalate) (PET). Scanning electron microscopy (SEM) is also employed as an auxiliary method to visualize the surface below the micron scale. The SEM images reveal a micron-sized ridgelike texture. Room temperature acid treatment makes the ridges become more pronounced, while treatment with boiling acid uncovers fiberlike structures of roughly 1 microm diameter. All samples display an apparent surface fractal dimension of Ds = 2.4 in the wave vector range 0.001-0.02 A(-1). Nitric acid at room temperature increases the surface oxygen content only by 3 at. %, while all the adsorption properties and structural parameters reported in this paper are virtually unaffected. Significant differences in the morphology at submicron scales appear only after boiling acid treatment. The resulting carbon remains highly microporous, but the loss of Brunauer-Emmett-Teller (BET) surface area from about 1150 to 304 m2/g is approximately 75%. In addition to the principal peak at around 8 A, fresh peaks appear in the polydisperse Horvath-Kawazoe (HK) pore size distribution owing to the burnoff of intervening walls. The average width of the slitlike pores calculated from the Dubinin-Radushkevich (DR) plot increases from 8.4 to 11 A. The minimum slit width where the applied probe molecules, that is, nitrogen and hexane, can enter increases from about 5 to about 5.4 A. The separation distance of the basic structural units is practically unchanged. When, however, this carbon is in contact with hexane, this distance expands from about 19 to 27 A. The swelling is consistent with the deformable nature of this sample also illustrated by the low-pressure hysteresis and the reduced helium density. Particular attention was paid to the surface areas derived from low-temperature nitrogen adsorption and X-ray measurements. Owing to the wide spatial range of the structures in these samples, estimates of the specific surface area of activated carbons can be substantially in error unless both upper and lower q ranges of the SAXS spectra are taken into account. Surface areas derived from the adsorption data either by the BET or the DR approaches were always below the values obtained by standard SAXS. As an example, the carbon sample functionalized at room temperature gave surface area values of 1114, 1293, and 1970 m2/g, respectively. The possibility that this difference is caused by inaccessible pores was excluded by contrast variation measurements with hexane. PMID- 15803715 TI - Thermal stability of perfluoroalkyl silane self-assembled on a polycrystalline aluminum surface. AB - Thermal stability behavior of 1H, 1H,2H,2H-perfluorooctyl trichlorosilane self assembled on aluminum substrates is characterized using a grazing-angle Fourier transform infrared spectrometer, Fourier transform-Raman spectroscopy, and contact angle measurements. The self-assembled monolayer (SAM) is heated quasi statically from room temperature to above 633 K with a heating rate of 1 K/s. Variations in peak frequencies, integrated areas of intensities of symmetric and antisymmetric CF2 stretches, and the relative tilt angle of the SAM are reported. We find that the conformational order in the SAM is not disrupted because of thermal cycling when the peak temperature is below 423 K. When the peak temperature is between 423 and 603 K, the cycling results in only a partial retention of the original order. When the peak temperature is above 603 K, the process is completely irreversible. Surface-enhanced Raman spectroscopy and contact angle measurements support these observations. We confirm these trends for a longer chain (1H,1H,2H,2H-perfluordecyl trichlorosilane) molecule of the same family using the same techniques. We discuss the possible reasons for the changes in light of the tilting-untilting and uncoiling-coiling of the helical silane monolayer. PMID- 15803716 TI - Comparative study of electrochemically directed assembly versus conventional self assembly of thioacetyl-terminated oligo(phenylene ethynylene)s on gold and platinum surfaces. AB - The assembly of thioacetyl-terminated oligo(phenylene ethynylene)s (OPEs) on Au and Pt surfaces under an electric potential (electrochemical assembly, EA) was compared to assembly at an open circuit (conventional self-assembly, CSA). Cyclic voltammetry and ellipsometry were used to characterize the adsorption kinetics of self-assembled monolayers formed by these two techniques. The adsorption rate of the EA was remarkably faster at positive potentials but slower at negative potentials than that of the CSA, The EA at 400 mV proceeded about 800 times faster than the CSA when exposed to the same solution concentrations. The adsorption rates of both EA and CSA were found to be dependent on the molecular structures of OPEs. OPEs containing electron-donating groups assemble faster than those with electron-withdrawing groups. The amount of time that the thioacetyl terminated OPE is in the presence of the base, for removal of the acetyl group to generate the thiolate, is called the deprotection time. Deprotection times play a critical role in achieving the maximum difference in adsorption rates between the EA and the CSA. The assembly must be initiated no later than 5 min after the basic deprotection is commenced so that the thiolate concentration remains low. The difference in the adsorption rates between EA and CSA might enable selective deposition of certain OPEs onto specific electrodes. PMID- 15803717 TI - Myoglobin/sol-gel film modified electrode: direct electrochemistry and electrochemical catalysis. AB - Direct electrochemical and electrocatalytic behavior of myoglobin (Mb) immobilized on carbon paste electrode (CPE) by a silica sol-gel film derived from tetraethyl orthosilicate was investigated for the first time. Mb/sol-gel film modified electrodes show a pair of well-defined and nearly reversible cyclic voltammetric peaks for the Mb Fe(III)/Fe(II) redox couple at about -0.298 V (vs Ag/AgCl) in a pH 7.0 phosphate buffer solution. The formal potential of the Mb heme Fe(III)/Fe(II) couple shifted linearly with pH with a slope of 52.4 mV/pH, denoting that an electron transfer accompanies single-proton transportation. An FTIR and UV-vis spectroscopy study confirms that the secondary structure of Mb immobilized on an electrode by a sol-gel film still maintains the original arrangement. The immobilized Mb displays the features of a peroxidase and acts in an electrocatalytic manner in the reduction of oxygen, trichloroacetic acid (TCA), and nitrite. In comparison to other electrodes, the chemically modified electrodes used in this study for direct electrochemistry and electrocatalysis of Mb are easy to fabricate and fairly inexpensive. Consequently, the Mb/sol-gel film modified electrode provides a convenient way to perform electrochemical research on this kind of protein. It also has potential use in the fabrication of bioreactors and third-generation biosensors. PMID- 15803718 TI - Atomic force microscopy study on the stability of a surface film formed on a graphite negative electrode at elevated temperatures. AB - The stability at elevated temperatures of a solid electrolyte interphase (SEI) formed on a graphite negative electrode in lithium ion batteries was investigated by storage tests and in situ atomic force microscopy (AFM) observation. When the fully discharged graphite electrode was stored at elevated temperatures, the irreversible capacity in the following cycle increased remarkably. On the other hand, when the electrode was stored at the fully charged state at elevated temperatures, it was severely self-discharged during storage. AFM observation of the SEI layer formed on a model electrode of highly oriented pyrolytic graphite revealed two important facts on the stability of the SEI at elevated temperatures: (i) dissolution and agglomeration of the SEI layer at the discharged state and (ii) serious SEI growth at the charged state. These phenomena well explain the results of the charge and discharge tests. It was also shown that the addition of vinylene carbonate greatly improves the stability of the SEI at elevated temperatures, and gives good charge and discharge performance after storage. PMID- 15803719 TI - Calcification of intraocular implant lens surfaces. AB - Calcification of octacalcium phosphate [Ca8H2(PO4)6 x 5H2O, OCP] on differently packaged "Ultem" and "Surefold" intraocular implant lens surfaces has been studied in vitro in solutions supersaturated with respect to OCP at pH = 7.10 and 37 degrees C. No mineral deposition was observed on the lenses packaged in Ultem vials even after treatment with behenic acid, one of the fatty acids identified on explanted lenses. Following treatment with behenic acid, nucleation of OCP occurred on the lenses from Surefold vials, which incorporate silicone gaskets; induction periods preceding calcification were about 6 h. No mineralization was found on the lenses in vials with other gasket materials, including polytetrafluoroethylene, fluorocarbon elastomer, and polypropylene. The results of this study indicate that both silicone and fatty acids such as behenic acid play important roles in inducing the in vivo calcification of OCP on IOL lenses; all of the lens treatment steps were necessary for nucleation induction. PMID- 15803721 TI - A nonviral transfection approach in vitro: the design of a gold nanoparticle vector joint with microelectromechanical systems. AB - Au nanoparticles modified with 21-base thiolated-oligonucleotides have been evaluated as delivery vehicles for the development of a nonviral transfection platform. The electromigration combined with electroporation for DNA delivery in an osteoblast like cell was employed to test on microchips. Electroporation introduces foreign materials into cells by applying impulses of electric field to induce multiple transient pores on the cell membrane through dielectric breakdown of the cell membrane. On the basis of the characteristic surface plasmon of the Au particles, UV-vis absorption was utilized to qualitatively judge the efficiency of delivery. Transmission electron microscopy images and atomic absorption measurements (quantitative analysis) provided evidence of the bare Au and Au/oligonucleotide nanoparticles before and after electroporation and electromigration function. The experiments demonstrated that electrophoretic migration followed by electroporation significantly enhanced the transportation efficiency of the nanoparticle-oligonucleotide complexes as compared with electroporation alone. Most interestingly, Au capped with oligonucleotides led to optimal performance. On the other hand, the bare Au colloidal suspensions resulted in aggregation, which might be an obstacle to the internalization process. In addition, analytical results demonstrated an increase in the local particle concentrations on the cell surface that provided additional support for the mechanism underlying the improved Au nanoparticle transportation into cells in the presence of electromigration function. PMID- 15803720 TI - Controlling mammalian cell interactions on patterned polyelectrolyte multilayer surfaces. AB - A newly discovered class of cell resistant surfaces, specifically engineered polyelectrolyte multilayers, was patterned with varying densities of adhesion ligands to control attachment of mammalian cells and to study the effects of ligand density on cell activity. Cell adhesive patterns were created on cell resistant multilayer films composed of poly(acrylic acid) and polyacrylamide through polymer-on-polymer stamping of poly(allylamine hydrochloride) PAH and subsequent reaction of the amine functional groups with an adhesion ligand containing RGD (Arg-Gly-Asp). These cell patterns demonstrated great promise for long-term applications since they remained stable for over 1 month, unlike ethylene glycol functional surfaces. By changing the stamping conditions of PAH, it was possible to alter the number of available functional groups in the patterned regions, and as a result, control the ligand density. Cell spreading, morphology, and cytoskeletal organization were compared at four different RGD densities. The highest RGD density, approximately 152 000 molecules/microm2, was created by stamping PAH at a pH of 11.0. Lowering the stamping ink pH led to patterns with lower ligand surface densities (83 000 molecules/microm2 for pH 9.0, 53,000 molecules/ microm2 for pH 7.0, and 25 000 molecules/microm2 for pH 3.5). An increasing number of cells attached and spread as the RGD density of the patterns increased. In addition, more cells showed well-defined actin stress fibers and focal adhesions at higher levels of RGD density. Finally, we found that pattern geometry affected cytoskeletal protein organization. Well-formed focal adhesions and cell-spanning stress fibers were only found in cells on wider line patterns (at least 25 microm in width). PMID- 15803722 TI - A molecular dynamics study of monolayers of nonionic poly(ethylene oxide) based surfactants. AB - Molecular dynamics simulations of monolayers of nonionic, poly(ethylene oxide) based surfactants are reported. Specifically, alcohol ethoxylates and alkylphenol ethoxylates are compared in terms of the varying architecture of the molecules for the development of a structure-behavior relationship. Interfacial density profiles are used to assess the structure of the monolayers, the penetration of water and oil into the monolayers, and the solvation of the hydrophiles and hydrophobes. Chain conformational descriptors are used to examine the molecular structure of the surfactants. The simulations revealed that monolayers of alcohol ethoxylates are considerably more diffuse than their alkylphenol counterparts, with the packing being governed by the size of the hydrophile. With the exception of the branched alcohol ethoxylate, the intermixing of the bulk phases within monolayers of alcohol ethoxylates increases with increasing hydrophile length. By comparison, the packing of alkylphenol ethoxylates within the monolayer is governed by the aromatic nucleus in the molecule. No specific interaction is observed between the aromatic rings of neighboring molecules. Monolayers of alkylphenol ethoxylates are more compact than their alcohol counterparts, resulting in more effective separation of the bulk water and oil phases. PMID- 15803723 TI - Interaction of cytolytic toxin CytB with a supported lipid bilayer: study using an acoustic wave device. AB - An acoustic technique was used to monitor the interaction of the pore-forming cytolytic toxin CytB with a positively charged supported lipid bilayer. The acoustic device, which is based on a waveguide geometry, is sensitive to changes in the mass of the supported bilayer. The specificity of the interaction, rate and extent of the association, reversibility and effect of previous depositions of toxin were investigated. The CytB was found to bind irreversibly to the lipids at all fractional coverages even when the protein-to-lipid ratio was high enough to imply that the protein was associating with the external surface of the bilayer. The CytB formed stable structures with the bilayer at high protein surface concentrations and did not appear to disrupt the bilayer in the manner of a detergent. The rate of association with the bilayer was found to be directly proportional to the solution concentration of CytB at higher concentrations but appeared to be low at a CytB solution concentration of 5 microg mL(-1), leading to relatively low amounts of CytB being associated with the bilayer. PMID- 15803724 TI - Study of molecular interactions between a phospholipidic layer and a pH-sensitive polymer using the Langmuir balance technique. AB - Molecular interactions between a terminally alkylated pH-sensitive N isopropylacrylamide copolymer DODA-poly(NIPAM-co-MAA) and a monolayer of distearoylphosphatidylcholine (DSPC) at the air/water interface are investigated using the Langmuir balance technique. The compression isotherms ofthe copolymer monolayer at the air-water interface confirm that the copolymer undergoes a structural transition with a change in pH ranging from an extended coil state at neutral pH to a collapsed globular state at a pH corresponding to the pH of the polymer phase transition. Adsorption kinetics of DODA-poly(NIPAM-co-MAA) in the DSPC monolayer is analyzed using a first-order kinetics model allowing an effective interaction area Ax between DSPC and DODA-poly(NIPAM-co-MAA) molecules to be evaluated. The results clearly indicate that the interaction area increases with a decrease in pH. The results also suggest that the penetration of the DODA poly(NIPAM-co-MAA) within the phospholipid monolayer is enhanced by a decrease in pH which causes a change in the copolymer structure and an increase in specific attractive interactions between the copolymer and the phospholipid. Therefore, the copolymer can trigger the destabilization or rupture of the phospholipidic layer through a simple variation in its structure associated with a variation in molecular interactions when coupled or inserted within the membrane. This study greatly supports the prospects of the copolymer-functionalized liposomes as stable and tunable carrier systems for in vivo applications in drug delivery. PMID- 15803725 TI - Effects of temperature, salt, and deuterium oxide on the self-aggregation of alkylglycosides in dilute solution. 1. n-nonyl-beta-D-glucoside. AB - The influence of salt, temperature, and deuterium oxide on the self-aggregation of n-nonyl-beta-D-glucoside (beta-C9G1) in dilute solution has been investigated by static and dynamic light scattering, neutron scattering, and tensiometry. Scattering data show that the micelles can be described as relatively stiff, elongated structures with a circular cross section. With a decrease of temperature, the micelles grow in one dimension, which makes it surprising that the critical micelle concentration (cmc) shows a concomitant increase. On the other hand, substitution of D2O for H2O causes a large increase in micelle size at low temperatures, without any appreciable effect on cmc. With increasing temperature, the deuterium effect on the micelle size diminishes. The effects of salt on the micelle size and cmc were found to follow the Hofmeister series. Thus, at constant salt concentration, the micelle size decreased according to the sequence SO4(2-) > Cl- > Br- > NO3- > I- > SCN-, whereas the effect on cmc displays the opposite trend. Here, I- and SCN are salting-in anions. Similarly, the effects of cations decrease with increasing polarizability in the sequence Li+ > Na+ > K+ > Cs+. At high ionic strength, the systems separate into two micellar phases. The results imply that the size of beta-C9G1 micelles is extremely sensitive to changes in the headgroup size. More specifically, temperature and salt effects on effective headgroup size, including intermolecular interactions and water ofhydration, are suggested to be more decisive for the micelle morphology than the corresponding effects on unimer solubility. PMID- 15803726 TI - Studies on the cellulose-binding domains adsorption to cellulose. AB - Cellulose-binding domains (CBD) are modular peptides, present in many glycanases, which anchor these enzymes to the substrate. In this work, the effect of CBD adsorption on the surface properties of a model cellulose, Whatman CF11, was studied. The methods applied include inverse gas chromatography (IGC), ESCA, X ray diffraction, and scanning electron microscopy (SEM). The CBD partition affinity (0.85 L/g) was calculated from adsorption isotherms. However, true adsorption equilibrium does not exist, since CBDs are apparently irreversibly adsorbed to the fibers. Both IGC and ESCA showed that fibers with adsorbed CBD have a lower acidic character and also a slightly higher affinity toward aliphatic molecules. This may however be a consequence of an increased surface area, a hypothesis that is supported by microscopic observations. The crystallinity index was not affected by CBD treatment. PMID- 15803727 TI - In situ study of colloid crystallization in constrained geometry. AB - We visualized in real time electrodeposition-driven colloid crystal growth on patterned conductive surfaces. The electrode was patterned with dielectric ribs and conductive grooves; the groove width was commensurate or incommensurate with a two-dimensional colloid crystal lattice. Electrodeposition was carried out against gravity to decouple sedimentation and electrodeposition of colloid particles. Our experiments reveal the following: (i) Colloid crystal growth occurs under the action of electrohydrodynamic forces, in contrast with colloid assembly under the action of capillary forces. (ii) Confinement of the colloid arrays reduces the size of particle clusters. Small clusters easily undergo structural rearrangements to produce close-packed crystals when the groove width is commensurate or nearly commensurate with the 2D lattice. (iii) Incommensurability between the two-dimensional crystalline lattice and the groove width exceeding ca. 15% leads to the formation of non-close-packed structures and the distortion of colloid arrays. PMID- 15803728 TI - Titania-coated polystyrene hybrid microballs prepared with miniemulsion polymerization. AB - Hybrid microballs with polystyrene cores coated by titania nanoparticles were prepared in miniemulsion polymerization. Acrylic acid was used as a comonomer to promote locating titania nanoparticles on the polymer's surface. The addition of a hydrophobic agent effectively prevents monomer diffusing into the aqueous phase. The morphology of hybrid particles was examined with the transmission electron microscope, and its change pattern with reactive conditions was observed. The infrared spectra of hybrid nanoparticles showed that there existed a certain interaction between titania nanoparticles and polymers. The crystallization morphology of hybrid particles before and after calcination was characterized with X-ray diffraction. PMID- 15803729 TI - Ultraporous single phase iron oxide-silica nanostructured aerogels from ferrous precursors. AB - Monoliths of iron oxide-silica aerogel nanocomposites have been synthesized using a novel synthesis route which consists of impregnating silica wet gels with anhydrous iron(II) precursors followed by ethanol supercritical drying of the gels. The process yields aerogels exhibiting high porosity, large surface areas (approximately 900 m2/g), rather low densities (approximately 0.6 g/cm3), and a homogeneous distribution of single-phase maghemite, gamma-Fe2O3, nanoparticles with average sizes in the 7-8 nm range. Remarkably, the gamma-Fe2O3 nanoparticles are obtained in the as-dried state without the need of postannealing. The nanoparticles are mostly superparamagnetic at room temperature but become blocked in a ferrimagnetic state at lower temperatures. PMID- 15803730 TI - Effect of surface modification on back electron transfer dynamics of dibromo fluorescein sensitized TiO2 nanoparticles. AB - Electron injection and back electron transfer (BET) dynamics have been carried out for dibromo fluorescein (DBF) sensitized TiO2 nanoparticles capped (modified) with sodium dodecyl benzene sulfonate using transient absorption techniques in picosecond and microsecond time domain. BET dynamics have been compared with bare (unmodified) nanoparticles for the same DBF/TiO2 system. It has been observed that BET reaction is slow on the modified surface compared to a bare surface in earlier time domain (picosecond). This observation has been explained by the fact that on surface modification the energy levels of the semiconductor nanoparticles are pushed up in energy. As a result, the free energy of reaction (-deltaG zero) for BET reaction of a dye/SM-TiO2 system increases as compared to the dye/bare TiO2 system. High exoergic BET reaction in dye-sensitized TiO2 nanoparticles surfaces fall in the Marcus inverted regime, so with increasing free energy of reaction, BET rate decreases on the modified surface. However, a reversible trend in BET dynamics has been observed for the above systems in the longer time domain (microsecond). In microsecond time domain BET reaction is faster on the modified surface as compared to on the bare surface. Modification of this surface reduces the density of deep trap states. Recombination dynamics between deep-trapped electron and parent cation is slow due to low coupling strength of BET reaction. As the density of deep-trapped electrons is high in bare particles, BET reaction is slow in longer time domain. PMID- 15803732 TI - Layer-by-Layer electrostatic self-assembly of polyelectrolyte nanoshells on individual carbon nanotube templates. AB - Carbon nanotubes have been featured prominently in the nanotechnology research for some time, yet robust strategies for noncovalent chemical modification of the nanotube surface are still missing. Such strategies are essential for the creation of functional device architectures. Here, we present a new general procedure for carbon nanotube modification based on polyelectrolyte layer-by layer assembly. We have built multilayer structures around individual carbon nanotube bridges by first modifying the nanotube surface with a pyrene derivative followed by layer-by-layer deposition of polyelectrolyte macroions on the nanotube. Transmission electron microscopy and scanning confocal fluorescence microscopy images confirm the formation of nanometer-thick amorphous polymer nanoshells around the nanotubes. These multilayer polyelectrolyte shells on individual carbon nanotubes introduce nearly unlimited opportunities for the incorporation of various functionalities into nanotube devices, which, in turn, opens up the possibility of building more complex multicomponent structures. PMID- 15803731 TI - Nonlithographic micro- and nanopatterning of TiO2 using polymer stamped molecular templates. AB - The polymer-on-polymer stamping technique was used to template patterned TiO2 onto polymer thin films. Polystyrene-b-polyvinyl pyridine diblock copolymer (PS-b PVP) was stamped on a layer-by-layer assembled thin film of poly(allylamine hydrochloride) and poly(acrylic acid). After rinsing the surface with a good solvent for the block copolymer, an adsorbed PS-b-PVP monolayer remained on the polyelectrolyte film, resulting in a pattern of alternating hydrophobic and carboxylic acid containing hydrophilic regions. The surface was used as a template for the selective deposition of TiO2 on the multilayer surface, using an acid-catalyzed hydrolysis of(NH4)2TiF6. Using this novel approach, we have successfully demonstrated the patterning of TiO2 film on a polyelectrolyte multilayer. Finally, nanoscale features consisting of 200 nm lines alternating with a 350 nm period was accomplished. This paper represents the first such attempt to create an all-polymer nonlithographic template for the directed deposition of TiO2 or related metal oxides; this technique, which utilizes the versatile polyelectrolyte multilayer process, enables the construction of complex polymer-inorganic microstructures suitable for electrooptical and photonic applications. PMID- 15803733 TI - Luminescent polymer microcapsules addressable by a magnetic field. AB - The simultaneous encapsulation of both luminescent semiconductor and magnetic oxide nanoparticles in polymer microcapsules is demonstrated for the first time. Highly luminescent CdTe semiconductor nanocrystals serve as luminescent markers, while magnetic Fe3O4 nanoparticles allow external manipulation of the capsules by magnetic field. The method introduced is general enough to allow the fabrication of different types of multifunctional capsules in a similar way. The use of multifunctional water-compatible capsules introduced in this paper for the controlled release and directed drug delivery in biological systems is envisaged. PMID- 15803734 TI - Moisture absorption and absorption kinetics in polyelectrolyte films: influence of film thickness. AB - Specular X-ray reflectivity (XR) and quartz crystal microbalance (QCM) measurements were used to determine the absorption of water into thin poly(4 ammonium styrenesulfonic acid) films from saturated vapor at 25 degrees C. The effect of film thickness on the absorption kinetics and overall absorption was investigated in the range of thickness from (3 to 200) nm. The equilibrium swelling of all the films irrespective of film thickness was (0.57+/-0.03) volume fraction. Although the equilibrium absorption is independent ofthickness, the absorption rate substantially decreases for film thickness < 100 nm. For the thinnest film (3 nm), there is a 5 orders of magnitude decrease in the diffusion coefficient for water. PMID- 15803735 TI - Tuning the interfacial properties of grafted chains with a pH switch. AB - Environmentally responsive, water-soluble polymers have a wide variety of uses ranging from drug delivery to viscosity modifiers. Their utility lies in the ability to use environmental perturbations to dramatically alter the material properties. Here, we describe the interfacial properties of a hydrophobically modified copolymer of N-isopropylacrylamide and glycinylacrylamide (NIPAM-N-Gly (C18)2), which is both temperature and pH responsive. Direct force measurements quantified the substantial pH-dependent change in the molecular properties of end grafted NIPAM-N-Gly-(C18)2 monolayers. At pH 8.0, where the glycine side chains are ionized, the polymers exhibit stereotypical polyelectrolyte behavior. Side chain neutralization at pH 5.0 causes a substantial decrease in the film thickness, and the polymer films adhere strongly. The adhesion is presumably through H-bonding between the glycine side chains. Our findings revealed the likely molecular basis of pH-dependent changes in the copolymer films and identified clear design criteria for tuning the interfacial properties of these polymer films. PMID- 15803736 TI - Self-assembly of gold nanoparticles on fullerene nanospheres. AB - A C60-pyrrolidine derivative with a hydrophobic-hydrophilic-hydrophobic structure (2-{3,4-di{2-[2-(2-decyloxyethoxy)ethoxy]ethoxy}}phenyl-3,4-fulleropyrrolidine, DTPF) has been synthesized and well-characterized. This compound could form stable nanospheres by simply injectingits tetrahydrofuran (THF) solution into water and then removing THF by purging gaseous nitrogen in sequence. Novel nanoassemblies of DTPF nanospheres and gold nanoparticles were obtained through in situ photoreduction of aqueous HAuCl4 in the presence of DTPF nanospheres, which were confirmed by UV-visible, transmission electron microscopy, X-ray powder diffraction, and X-ray photoelectron spectroscopy methods. It is proposed that the interaction between the positively charged nitrogen atom and the gold nanoparticles is the main driving force for the formation of the nanoassemblies. PMID- 15803737 TI - Interaction of asphaltenes with nonylphenol by microcalorimetry. AB - This paper shows the work performed in the study of the capability of isothermal titration calorimetry (ITC) to characterize the interaction between petroleum asphaltenes with a model molecule, namely, nonylphenol. ITC is widely used in biochemistry to study the interaction of proteins with ligands. The intention is to transfer the knowledge into the asphaltene field, with the aim of getting a better understanding of the mechanism of interaction, as well as the energies involved in this process. Calorimetric experiments show that nonylphenol has a complex mechanism of interaction with asphaltenes in toluene, including more than one process. Several models have been used to fit the experimental data. The enthalpies calculated with a model based on polymerization are in the order of -1 to -7 kJ/mol, which are very close to the hydrogen bond energies. This shows the capability of ITC to provide experimental data to the modeling of asphaltene behavior. The number of sites of interaction has been inferred by means of a model taken from protein-ligand science. The values obtained are in the range two to five sites per molecule, assuming an average Mw of 1000 units. PMID- 15803739 TI - Graphoepitaxial deposition of cationic polymer micelles on patterned SiO2 surfaces. AB - We have studied the deposition of polymer micelles formed from poly(styrene) block-poly(2-vinylpyridine) (PS-PVPH+) from room-temperature aqueous solutions at pH 1 onto a hydrophilic Si/SiO2 surface with a relief pattern 100 nm deep with variable widths. It has been found that the micelle density is substantially higher and the ordering of the micelles is improved for micelles that adsorb in the 100 nm depressions in the width range of ca. 500-5000 nm. We ascribe these effects to capillary forces that pull the aqueous solution into the canyons where the micelles can be trapped. While the ordering of the micelles can be substantial, they do not form a perfect hexagonal crystal. If the surface is chemically modified by a Au coating, the micelle-surface interaction is strengthened and the degree of ordering is diminished. These results demonstrate that a combination of graphoepitaxy and processing conditions (speed of substrate withdrawal or evaporation of solvent) can be used to make fairly ordered polymer micelle arrays over a space of (at least) several millimeters. PMID- 15803738 TI - Covalent attachment of phospholipid analogous polymers to modify a polymeric membrane surface: a novel approach. AB - A novel method for the surface modification of a microporous polypropylene membrane by tethering phospholipid analogous polymers (PAPs) is given, which includes the photoinduced graft polymerization of N,N-dimethylaminoethyl methacrylate (DMAEMA) and the ring-opening reaction of grafted poly-(DMAEMA) with 2-alkyloxy-2-oxo-1,3,2-dioxaphospholanes. Five 2-alkyloxy-2-oxo-1,3,2 dioxaphospholanes, containing octyloxy, dodecyloxy, tetradecyloxy, hexadecyloxy, and octadecyloxy groups in the molecular structure, were used to fabricate the PAP-modified polypropylene membranes. The attenuated total reflectance FT-IR spectra of the original, poly(DMAEMA)-grafted, and PAP-modified membranes confirmed the chemical changes on the membrane surface. Scanning electron microscope pictures showed that, compared with the original membrane, the surface porosities ofpoly(DMAEMA)-grafted and PAP-modified membranes were somewhat reduced. Water contact angles measured by the sessile drop method on PAP-modified membranes were slightly lower than that on the original polypropylene membrane, but higher than those on poly(DMAEMA)-grafted membranes with the exception of octyloxy-containing PAP-modified membranes. However, BSA adsorption experiments indicated that the five PAP-modified membranes had a much better protein resistant property than the original polypropylene membrane and the poly(DMAEMA) grafted membranes. For hexadecyloxy- and octadecyloxy-containing PAP-modified membranes, almost no protein adsorption was observed when the grafting degree was above 6 wt %. It was also found that the platelet adhesion was remarkably suppressed on the PAP-modified membranes. All these results demonstrate that the described approach is an effective way to improve the surface biocompatibility for polymeric membranes. PMID- 15803740 TI - Thermally activated nanowear modes of a polymer surface induced by a heated tip. AB - We report on nanoscale wear induced by atomic force microscopy using a heated cantilever/tip on a 20 nm thick film of polystyrene. Wear modes dependent on tip temperature characteristics were identified. Below the glass transition temperature of the polymer, the formation of quasi one-dimensional surface ripples with a typical period on the order of 100 nm was observed. We found that the ripple height typically saturates at 20 nm and that the buildup rate depends on temperature and load. From these observations we can calculate an activation energy for the ripple formation, which is on the order of 0.4 eV, at temperatures close to but below the glass transition temperature of the polymer. In the glass transition regime the ripple formation is strongly enhanced. An abrupt change of the wear mode is observed as the polymer in contact with the tip is heated above the glass transition temperature. Here, polymer material is transported along with the propagating tip. PMID- 15803741 TI - Nanoscale colloids in a freely adsorbing polymer solution: a Monte Carlo simulation study. AB - A key issue in nanoscale materials and chemical processing is the need for thermodynamic and kinetic models covering colloid-polymer systems over the mesoscopic length scale (approximately 1-100 nm). We have applied Monte Carlo simulations to attractive nanoscale colloid-polymer mixtures toward developing a molecular basis for models of these complex systems. The expanded ensemble Monte Carlo simulation method is applied to calculate colloid chemical potentials (micro(c)) and polymer adsorption (gamma) in the presence of freely adsorbing Lennard-Jones (LJ) homopolymers (surface modifiers). gamma and micro(c) are studied as a function of nanoparticle diameter (sigma(c)), modifier chain length (n) and concentration, and colloid-polymer attractive strength over 0.3 < Rg/sigma(c) < 6 (Rg is the polymer radius of gyration). In the attractive regime, nanocolloid chemical potential decreases and adsorbed amount increases as sigma(c), or n is increased. The scaling of gamma with n from the simulations agrees with the theory of Aubouy and Raphael (Macromolecules 1998, 31, 4357) in the extreme limits of Rg/sigma(c). When Rg/sigma(c) is large, the "colloid" approaches a molecular size and interacts only locally with a few polymer segments and gamma approximately n. When Rg/sigma(c) is small, the system approaches the conventional colloid-polymer size regime where multiple chains interact with a single particle, and gamma approximately sigma(c)2, independent of n. In contrast, adsorption in the mesoscopic range of Rg/sigma(c) investigated here is represented well by a power law gamma approximately n(p), with 0 < p < 1 depending on concentration and LJ attractive strength. Likewise, the chemical potential from our results is fitted well with micro(c) approximately n(q)sigma(c)3, where the cubic term results from the sigma(c) dependence of particle surface area (approximately sigma(c)2) and LJ attractive magnitude (approximately sigma(c)). The q-exponent for micro(c) (micro(c) approximately n(q)) varies with composition and LJ attractive strength but is always very close to the power exponent for gamma (gamma approximately n(p)). This result leads to the conclusion that in attractive systems, polymer adsorption (and thus polymer colloid attraction) dominates the micro(c) dependence on n, providing a molecular interpretation of the effect of adsorbed organic layers on nanoparticle stability and self-assembly. PMID- 15803742 TI - Extremes of some foam properties and elasticity of thin foam films near the critical micelle concentration. AB - The elasticity of open and closed thin foam films is analyzed. The elasticity modulus of a closed film is shown to be additive with respect to contributions from Gibbs elasticity and disjoining pressure. A detailed expression for the film elasticity modulus explains the pronounced maxima of foaminess and foam stability near the critical micelle concentration observed earlier in many experiments. A theory of transversal elasticity of thin foam films is formulated under conditions excluding the action of Gibbs elasticity. Near the critical micelle concentration, the theory predicts maxima of the transversal elasticity modulus and of the films thickness as functions of concentration at a given disjoining pressure. The prediction has been verified experimentally by measuring the film thickness in equilibrium foam as a function of height. PMID- 15803743 TI - Photoinduced cleavage of alkyl monolayers on Si. PMID- 15803745 TI - Flow-controlled vertical deposition method for the fabrication of photonic crystals. PMID- 15803744 TI - Morphology control of PbWO4 nano- and microcrystals via a simple, seedless, and high-yield wet chemical route. PMID- 15803746 TI - Influence of the thiol position on the attachment and subsequent hybridization of thiolated DNA on gold surfaces. PMID- 15803747 TI - Microwave-assisted preparation of hierarchical mesoporous-macroporous boehmite AlOOH and gamma-Al2O3. PMID- 15803748 TI - Detection of mediator-induced airway constriction by barometric plethysmography in mice. AB - The barometric method has recently been employed to detect airway constriction in small animals. This study was designed to evaluate the barometric method to detect mediator-induced central and peripheral airway constriction in BALB/c mice. First, the central airway constrictor carbachol and the peripheral airway constrictor histamine were employed to induce airway constriction, which was detected by both the conventional body plethysmography and the barometric method in anesthetized mice. Second, bronchoconstriction induced by aerosolized carbachol or other mediators was detected with the barometric plethysmography in conscious, unrestrained mice. Carbachol inhalation caused about four-fold increase in pulmonary resistance (RL) and about two-fold increase in enhanced pause (Penh) in anesthetized mice. In contrast, in the same preparation, histamine aerosol induced a decrease in dynamic compliance (Cdyn), with no alteration in RL or Penh. In awake mice, carbachol and methacholine caused increases in Penh, frequency, and tidal volume (VT). On the other hand, histamine, histamine + bradykinin, and prostaglandin-D2 did not alter Penh but decreased VT in conscious mice. These data suggest that there was no sufficient evidence to indicate that Penh could be a good indicator of bronchoconstriction for the whole airways. PMID- 15803749 TI - Effects of juice from Morinda citrifolia (Noni) on gastric emptying in male rats. AB - The effects of juice from Morinda citrifolia (noni) on gastric emptying, gastrointestinal transit, and plasma level of cholecystokinin (CCK) in rats were studied. Male rats were given noni by gavage at levels of 0.25, 1, or 4 ml/kg once per day for one or 7 days. The rats in the control group were given water, while the rats in the experimental group were fasted overnight before measurement of gastrointestinal motility. Gastrointestinal motility was assessed in rats 15 min after intragastric instillation of a test meal containing charcoal (10%) and Na251CrO4 (0.5 microCi/ml). Gastric emptying was determined by measuring the amount of radiolabeled chromium contained in the small intestine as a percentage of the initial amount received. Then, gastrointestinal transit was evaluated by calculating the geometric center of distribution of the radiolabeled marker. Finally, blood samples were collected for measurement of CCK by radioimmunoassay. The administration of noni at 0.25 ml/kg, but not at 1 ml/kg and 4 ml/kg, for 1 day significantly inhibited gastric emptying. In contrast, gastric emptying was significantly inhibited by oral noni (0.25, 1, or 4 ml/kg) for 7 days. Intraperitoneal injection of lorglumide (5 or 10 mg/kg), a selective CCK1 receptor antagonist, effectively attenuated the noni-induced inhibition of gastric emptying. The intestinal transit and body weight, food intake, water intake, urine volume as well as feces weight were not altered by the administration of noni either acutely or chronically, but the administration of oral noni (1 ml/kg) for 7 days increased the level of plasma CCK in male rats. These results suggest that oral noni inhibits gastric emptying in male rats via a mechanism involving stimulation of CCK secretion and CCK1 receptor activation. PMID- 15803750 TI - Pentagastrin-induced gastric acid secretion in the diabetic rats: role of insulin. AB - Streptozotocin-induced diabetic rats have excessively pentagastrin-simulated acid output in which insulin seems to attenuate rather than further stimulate acid output. The aim of this study was to determine the insulin impact on pentagastrin stimulated acid output of diabetic and non-diabetic rats to resolve whether an attenuated effect does exist. Diabetic rats were induced by the streptozotocin i.v. injection four days before acid study. Some streptozotocin-treated rats additionally received daily insulin (2.4 IU/kg) injection. Using an autotitrator, acid output was measured every five minutes by the titration of gastric perfusate. Basal output was collected for 45 min before the 90-min pentagastrin infusion (0.89 microg/kg/min). Plasma gastric inhibitory polypeptide (GIP) levels were measured. Both doses (0.067 and 0.133 IU/kg/min) of insulin infusion resulted in stimulated acid output in normal rats. The subsequent insulin infusion (0.133 IU/kg/min) for non-diabetic rats undergoing pentagastrin treatment suppressed their stimulated acid output almost down to the basal level. Pentagastrin-stimulation led to the excessively increased acid output of diabetic rats throughout the whole infusion period (P < 0.01). Correction of hyperglycemia with insulin for diabetic rats normalized the stimulated acid output. Measured basal and stimulated plasma GIP levels of those diabetic rats during acid stimulation remained higher, regardless of insulin treatment (P < 0.05). Our results suggest that insulin has the ability to attenuate pentagastrin-stimulated acid output in rats, whereas GIP is not involved in this attenuation. This effect appears to be responsible for the excessive acid output of diabetic rats undergoing pentagastrin stimulation. PMID- 15803751 TI - Estrogen ameliorates Nomega-nitro-L-arginine methyl ester-induced blood pressure increment in male spontaneously hypertensive rats: the role of cGMP. AB - Estrogen (17beta-estradiol, or E2) reduces systolic blood pressure (SBP) increment and increases aortic cyclic guanosine monophosphate (cGMP) in male spontaneously hypertensive rats (SHRs). It is unknown, however, whether the E2 enhanced aortic cGMP is essential for the BP-lowering effect or not. Nomega-nitro L-arginine-methyl ester (L-NAME), an L-arginine analogue and nitric oxide (NO) synthase inhibitor, significantly increases SBP and decreases aortic cGMP in male SHRs. We thus treated male SHRs with vehicle (corn oil) or E2 (s.c, 2 mg/kg/week) with or without L-NAME (20 mg/dl in the drinking water). SBP was measured weekly. Plasma nitrate/nitrite (NOx) concentrations and aortic cGMP levels were all measured at the end of the study. We found that SBP increment was significantly higher in L-NAME group, compared with the controls, and that E2 treatment reduced this L-NAME effect. Plasma 4NOx concentrations were not significantly different among different groups. Basal and acetylcholine-induced aortic cGMP, but not sodium nitroprusside-induced cGMP, were significantly lower in L-NAME group, compared with the controls. E2 co-administration did not modify L-NAME-induced aortic cGMP decrease. These data indicate that E2-induced BP-lowering effect in L NAME treated male SHRs is not closely associated with the enhancement of vascular cGMP. PMID- 15803752 TI - In vivo inhibition of inducible nitric oxide and evaluation of the brain tissue damage induced by Toxocara canis larvae in experimentally infected mice. AB - Nitric oxide (NO) is known to be produced by macrophages, endothelial cells and neurons and synthesized by an enzyme called nitric oxide synthase (NOS). Various effector mechanisms and infections can affect the NO production. Excessive amount of NO will lead to biochemical reactions, which cause toxic effects. In this study the role of NO has been evaluated in larval toxocarosis, which is a systemic parasite infection caused by T. canis larvae. Infection was established in the Balb/c mice with or without inducible NOS (iNOS) inhibition and the effects of infection and NOS inhibition were observed according to the results of SOD and LPx measurements in brain tissue and NADPH-diaphorase (NADP-d) histochemistry. Results of NADPH-d histochemistry indicate that iNOS inhibition has protective effect on the brains of infected mice and that larval T. canis infection could be related to oxidative stress, and NO production and iNOS inhibition can protect the tissue from damage in this infection. PMID- 15803753 TI - Anti-inflammatory effect of hydroxyethylpuerarin on focal brain ischemia/reperfusion injury in rats. AB - The objective of this study is to investigate the anti-inflammatory effect of hydroxyethylpuerarin on focal brain ischemia injury in rats and to explore its mechanisms of action. After 24 h of reperfusion following 2 h of cerebral ischemia, the infiltration of neutrophils was observed by myeloperoxidase (MPO) activity determination, the expression of intercellular adhesion molecule-1 (ICAM 1) was observed by western blot and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, and the nuclear translocation and DNA binding activity of nuclear factor-kappaB (NF-kappaB) were observed by western blot and electrophoretic mobility shift assay (EMSA). The results showed that hydroxyethylpuerarin could obviously inhibit the MPO activity and ICAM-1 expression following 2 hours of ischemia with 24 hours of reperfusion. The nuclear translocation and DNA binding activity were also decreased by hydroxyethylpuerarin treatment. These results suggested that hydroxyethylpuerarin could inhibit neutrophil-mediated inflammatory response after brain ischemia reperfusion in rats. This effect may be mediated by down-regulation of ICAM-1 and NF-kappaB activity. PMID- 15803754 TI - Activation and up-regulation of phospholipase D expression by lipopolysaccharide in human peripheral T cells. AB - In a previous study, we showed that bacterial LPS activates protein kinase C (PKC) and causes an intracellular pH (pHi) increase, but does not elevate intracellular calcium ([Ca2+]i) in human peripherial T cells. Hence this study aimed to investigate whether the activation of PKC was resulted from phospholipase D (PLD) catalysis by LPS. The activity of PLD was measured by the production of 3H-phosphatidylethanol from phosphatidic acid (PA), and the expression of PLD or IL-2 Ralpha was determined by reverse transcription polymerase chain reaction (RT-PCR) analysis. Enzyme-linked immunosorbent assay (ELISA) was used to analyze IL-2 and IL-4. Phytohemagglutinin (PHA), and phorbol 12-myristate 13-acetate (PMA) were used as controls. Our results indicated that (1) LPS-stimulated pHi elevation was PKC dependent; (2) After 30 min stimulation, LPS increased PLD activity via a measured production of 3H-phosphatidylethanol from phosphatidic acid and the initiation of PLD1a mRNA expression started; (2) LPS stimulated IL-2 R expression but not IL-2 and IL-4 secretion. Our findings suggested that the stimulation of PLD activity and its mRNA expression by LPS might be required for IL-2 R expression and a sustained PKC dependent pHi elevation but not for the secretion of IL-2 or IL-4 in human T cells. This indicated that LPS might enhance T cell adaptive immunity to resist Gram-negative bacterial infection. PMID- 15803755 TI - [The ethical minimum of animal protection--one of the effects of the German national objective "Ethical Animal Protection" on the legislature]. AB - In 2002 the current legal concept for animal protection "Ethischer Tierschutz" (ethics based animal protection) was declared a national objective in the Federal Republic of Germany. One of the aims laid down in the bill is to assure an ethical minimum (ethisches Mindestmass). An updating of and 7 Section 3 and and 17 No. 2b of the German Animal Welfare Act in compliance with the constitution and EU legislation could abolish the legal exception (regarding animal experiments) to the ban on inflicting "longer periods or repeated actions of severe pain or suffering". In this way an overall limit for causing severe harm to animals in any type of use, which is strongly demanded on the part of ethics, would be established, and this would equal an ethical minimum. PMID- 15803756 TI - [Plasma amino acid concentrations in aggressive dogs]. AB - Following the hypothesis that metabolic screens may be useful tools in the diagnosis of canine aggression we have investigated the blood plasma amino acid levels of dogs which have been found aggressive (N = 10) against dogs or men in comparison to non-aggressive dogs (N = 10). In summary, the aggressive dogs showed elevated plasma concentrations of the neurophysiological active aromatic amino acids tryptophan (46/171 micromol/l, p < 0,001), tyrosine (38/67 micromol/l, p < 0.01) and histidine (74/91 micromol/l, p < 0.01) and lower lysine concentrations (175/151 micromol/l, p < 0.05), which seems to point to a stress situation of these dogs. The nitrogen metabolism is impaired in the urea-cycle in the conversion of ornithine (17/34 micromol/l, p < 0.01) to citrulline (64/47 micromol/l). Higher levels of branched chain amino acids, especially leucine (122/150 micromol/l, p < 0.01), mainly metabolized in muscles, and isoleucin (60/71 micromol/l, p < 0.05) show a high energy potential. The acidose-stimulator methionine (48/78 micromol/l, p < 0.01) proved elevated. The results show that the changed behavior in the aggressive dogs is also reflected in their free amino acid plasma concentrations, independent of the question whether these data are the cause or the result of the aggressivity. PMID- 15803757 TI - [Placebo-controlled blinded study of postoperative pain therapy with carprofen and levomethadone in dogs with fractures]. AB - It was the aim of this placebo-controlled study to evaluate the analgesic efficacy of the NASAID carprofen and the pure m-agonist levomthadone over a five day postoperative evaluation period in dogs with fractures of the humerus or the femur (n = 30). Pain and sedation evaluation was carried out with a visual analogues system (VAS) and with the aid of a numerical estimation scale(NRS). The degree of lameness, the pain treshhold, the glucose and cortisol concentration curves as well as the respiration and heart rate and the systolic blood pressure were used as further pain indicators and to identify drug side effects. The levomethadon group displayed the lowest degree of pain on postoperative examination on the first day. On days 2 to 5, the carprofen group showed the lowest degree of pain in comparison to the placebo group. The levomethadon- and the carprofen group showed no statistically proven differences from day 2 on. Due to great variations in the pain scores and comparatively high median pain score especially on the first day of this study, the efficacy of all analgesics evaluated here must be regarded as insufficient in many cases. Only the parameter nociceptive pain treshhold showed a little, the degree of lameness, the glucose and cortisol levels showed no close correlation to the VAS and NRS pain scores and were therefore of little usefulness as postoperative pain indicators. No relevant clinical side effects caused by the used analgesics were detected in the kidney, the liver, the gastrointestinal tract and the circulatory system in this study. Rather, traumatically induced elevation of enzyme levels improves or normalised until the 5th day of the study. In addition, no negative effect on wound healing was noted, especially for carprofen. Therefore, the evaluated analgesics seems to be adequate for postoperative pain therapy also in fracture patients (trauma patients). However, the efficacy of all analgesics evaluated here must be regarded as insufficient in many cases. PMID- 15803758 TI - [Detection and species-specific differentiation of pestiviruses using real-time RT-PCR]. AB - An important prerequisite for an efficient eradication of pestiviruses like bovine viral diarrhea virus or classical swine fever virus are sensitive and specific detection methods. Beside antigen detection with antigen capture ELISAs and virus isolation using cell culture, the detection of virus genomes by reverse transcriptase polymerase chain reaction (RT-PCR) becomes more and more important. By using real-time RT-PCR, the disadvantages of conventional PCR methods concerning the risk of contamination and missing specificity controls are minimized. In the following, the validation and application of different real time RT-PCR systems for the detection of pestiviruses are presented. In addition, a new "panpesti" probe was tested, and the ability to differentiate between the major species of pestiviruses with known PCR protocols was determined using the Light Cycler PCR machine. PMID- 15803759 TI - [Bacterially conditioned thromboembolism in dairy cows--a retrospective study of 31 necropsy cases with special consideration of the causative complex]. AB - A study of 340 necropsied dairy cattle from northern Bavaria resulted in 31 animals (9%) showing evidence of pyemic thromboembolism. The most frequent pathomorphologic lesions consisted in endocarditis valvularis thromboticans of the tricuspid and/or mitral valve (21), embolic pneumonia (26), nephritis (13) and renal infarction (12). The most common isolate found in bacteriologic culture was Arcanobacterium pyogenes in 26 of 31 cases, followed by Staphylococcus aureus (2), Mannheimia haemolytica (2) and Streptococcus bovis (1) in rare cases. In 27 of 31 cows a possible cause of pyemic thromboembolism was found; in 23 cases claw diseases and decubital ulcera were probably responsible. PMID- 15803760 TI - [Epidemiological investigations of bovine Chorioptes mange in Germany]. AB - Chorioptic mange in cattle has long been recognised as a common external parasitosis in livestock. In order to evaluate the economic importance of Chorioptes bovis-mange, the seasonal prevalence in several types of husbandry in the states of Germany, location of clinical skin symptoms, procedure of diagnosis, influence on milk or meat production, and treatment especially in dairy farming, 1190 German large animal practicioners have been asked by questionnaires from which 367 responded. This paper gives an update on important epidemiological investigations on bovine chorioptic mange due to this inquiry. Since more than one third of the questioned veterinarians see tail mange often, it is of high importance for farm cattle. Chorioptes mange mainly occurs in dairy cattle stocks, and has especially negative effects on milk production and meat productivity. For treatment of this external parasitosis mainly macrocyclic lactons and pyrethroids are used. PMID- 15803761 TI - [Influence of inbreeding on litter size and the proportion of stillborn puppies in dachshunds]. AB - In the present study, factors influencing the litter size of puppies and the number and the proportion of stillborn puppies per litter were analysed in normally sized German dachshounds. Data was available for litters registered from 1987 to 2002 in the German dachshound kennel club. The records studied here contained 42,855 litters. The mean litter size was 5.22 +/- 2.01 puppies. The proportion of puppies born dead per litter was 6.6%. The age of dam, the puppy's year and month of birth, the hair type and the inbreeding coefficients of the puppies, the sire and the dam were significant on the litter size and the number and the proportion of puppies born dead. The litter size decreased and the percentage of puppies born dead increased with increasing inbreeding coefficients of the puppies, dams and sires. The increase of the inbreeding coefficient of the puppies by 1% led to a 0.06% higher percentage of puppies born dead per litter. The increase of the inbreeding coefficients of the dam and the sire by 1% resulted in 0.02% and 0.007% higher percentages of puppies born dead. Young and older dams had smaller litter size than middle-aged dams. The percentage of puppies born dead per litter increased with an increasing age of the dam. In the recent birth years, the litter size and the percentage of puppies born dead increased in German dachshounds. PMID- 15803762 TI - [Effects of a single dose of dexamethasone-21-isonicotinate on the phagocytosis performance of neutrophilic granulocytes and monocytes in periparturient dairy cows]. AB - The objective of the study was to describe the effects of a short acting glucocorticoid which was administered immediately after parturition in dairy cows on haematological, immunologic and metabolic parameters. The study was performed on 104 cows, 52 cows received an intramuscular injection of 2 mg/100 kg BW dexamethasone within 12 hours after parturition. The numbers and the phagocytic activity of granulocytes and monocytes were measured on day 1 and day 3 post partum in 17 randomly chosen cows from each group. From day 1 to day 3 the percentage of phagocytic granulocytes increased in the dexamethasone group from 73.2 +/- 6.1% to 79.6 +/- 6.6% (P < 0.05); in the control group the percentage remained unchanged (74.7 +/- 6.3% and 76.2 +/- 5.8%). The cellular phagocytic activity of granulocytes did not change significantly between day 1 and 3. In both groups the phagocytic activity of monocytes also remained unchanged. The concentrations of endotoxin, haptoglobin and the anti-lipid-A-IgG-antibody-titer did not show significant differences between the groups. Between both groups differences were not observed in the clinical parameters, milk yield or in the incidence of diseases in the peripartal period. The effects of the glucocorticoid on the metabolic parameters were especially seen in bilirubin and free fatty acid concentration. The present study shows that a single injection of glucocorticoids within 12 hours after parturition did not influence the phagocytic activity of neutrophils and monocytes negatively and did not have an effect on the measured parameters of acute-phase-response. Therefore, it seems possible to benefit from the positive effects of glucocorticoids on metabolism in this critical early postpartal period in high yielding cows. PMID- 15803763 TI - [Body and claw measurements as well as pressure distribution under the claws in heifers of different cattle breeds]. AB - In 56 heifers of the three breeds German Holstein (GH), German Brown (GB) und German Red (GR) the body weight and height were determined at the age of 350 to 550 days in intervals of 6 to 8 weeks. At the right front limb and the left hind limb the claw measurements dorsal wall length, diagonal length, heel length and height, the angle of the dorsal wall and the hardness of the claw were taken. Furthermore, the punctual pressure under the medial and the lateral claw of the right front limb and the left hind limb were determined using an electronic measuring system. The GH heifers weighing 387.3 kg on an average and having a mean height of 132.3 cm at the sacral bone in the course of the present study were the heaviest and tallest breed at the age of 12 to 18 months. GB had a weight of 366.2 kg and 106.2 cm height at the sacral bone. The GR were the smallest breed (126.0 cm) and weighed 337.7 kg. The heel at the front limb was on the average 1 cm shorter than at the hind limb and it was 0.5 cm lower. The angle of the dorsal border was 4-5 degrees higher at the hind limb. The GR had the flattest claws. The animals showed a larger average weight load on the medial claws than on the lateral claws especially of the front limbs. The highest pressures per cm2 were found underneath the rear part of the medial claw in all breeds. At the front limb, the GB showed significantly higher average pressures per cm2 under the front sectors of the claws than GH and GR, while the other values did not differ significantly between the breeds. The GH tended to have lower average pressures per cm2 underneath the claws of the hind limb, but this was only significant at the front medial sector of the claw. PMID- 15803764 TI - [Measurement of the elastic properties of the cancellous bone in the femoral head of the dog]. AB - The degenerative wear and pathologic damage of the joints are reasons for total endoprotheses in man as well as in dogs. The main problem is the aseptic loosening of the protheses. By usig the finite-element-method, the total endoprothesis is designed with new features, with the purpose of preventing loosening and being better adapted to load transmission. In order to simulate the femur of the dog for the numerical analysis, a material law is developed. By taking into account the anisotropy and the local density of the cancellous bone in the femoral head, the young's modules are experimentally determined. The measurements are performed by ultrasonic methods on femoral heads of euthanised dogs. The results show planar isotropic cancellous bone. PMID- 15803766 TI - Little foot and big thoughts--a re-evaluation of the Stw573 foot from Sterkfontein, South Africa. AB - The part of the fossil assemblage Stw573 consisting of some medial foot bones was initially reported by Clarke & Tobias (Science 269 (2002) 521). They found it to have both ape- and human-like qualities, being human-like proximally and ape-like distally. We have undertaken a re-examination of this pedal assemblage using a multivariate analysis; while we also found ape- and human-like qualities, they are in direct conflict with the original findings of Clarke and Tobias. We report an essentially ape-like morphology proximally and a human-like morphology distally; the talus and navicular were found to be ape-like and the medial cuneiform human-like. We also undertook a morphometric analysis of the medial cuneiform from the fossil assemblage OH8, as this was not included in the original OH8 study of Kidd et al (J Hum Evol 31 (1996) 269); this cuneiform was found to have a human-like morphology. Thus, the medial column findings from the two assemblages are very similar. This finding, coupled with the re-evaluation of the stratigraphy at Sterkfontein (Am J Phys Anthrop 119 (2002) 192), suggests that the two may have been contemporaneous. We also note that three broad patterns of modification have been identified, equating to proximal-distal lateral-medial (cranio-caudal) and dorsal plantar (posterior-anterior). It has not escaped our notice that these patterns are each controlled by specific genes or growth factors; we thus see a morphometric expression of our developmental past. PMID- 15803765 TI - [Low flow anaesthesia with isoflurane in the dog]. AB - The aim of the present study was to compare the safety of two low flow (LF) regimes [fresh gas flow (FGF) 20 ml/kg/min (group 2) and 14 ml/kg/min (group 3)] with the high flow (HF) technique (FGF 50 ml/kg/min; group 1) of isoflurane anaesthesia. Data were gathered from ninety dogs assigned for surgery under general anaesthesia with an expected duration of 75 minutes or longer. All dogs had an anaesthetic induction with 0,6 mg/kg I-methadone (maximum 25 mg) and 1 mg/kg diazepam (maximum 25 mg) i.v. Anaesthesia was maintained with isoflurane in a mixture of 50% O2 and 50% N2O as carrier gases, with controlled ventilation. The Monitoring included electrocardiogramm, body temperature, the temperature of in- and exspired gases, arterial oxygen saturation, arterial blood pressure as well as a continuous monitoring of inhaled and exhaled gas concentrations (O2, N2O, CO2, isoflurane). The consumption of isoflurane and carrier gases as well as the recovery times were evaluated for the three groups. The inspired oxygen concentrations always ranged above the minimum value of 30 Vol.-% during low flow anaesthesia. The arterial oxygen saturation ranged between 92-98%, the end tidal concentration of CO2 between 35 and 45 mmHg. Heart rate and arterial blood pressure were within normal limits. Recovery time was significantly shorter after LF than after HF anaesthesia. The highest decrease in body temperature occurred in the HF group 1 because of a significantly lower anaesthetic gas temperature. Despite this, LF anaesthesia resulted in a reduced consumption of carrier gases and volatiles. In conclusion, low flow anaesthesia with isoflurane is a safe technique and offers substantial economic advantages over high flow techniques and is moreover better tolerated by the patients. PMID- 15803767 TI - Sexual dimorphism and discriminant function sexing in indigenous South African crania. AB - This study aimed to examine sexual dimorphism in, and to produce a practical discriminant function for determining the sex of indigenous, Bantu-speaking, South African crania. The types of data to be used were a small number of traditional, or mathematically transformed three-dimensional, linear measurements, comparable to those in use by most physical and forensic anthropologists. The samples to be examined, separately and pooled, were of the Cape Nguni, Natal Nguni and Sotho subgroups. In addition, three local populations ('tribes'--Zulu, Xhosa and Southern Sotho) within these subgroups were also studied. Univariate male/female ratios indicate significant sexual dimorphism in the pooled South African crania. Canonical variates analysis of the pooled sample showed that facial width is the strongest discriminating morphometric variable; cranial length and basi-bregmatic height are the next most significant features. Eight measurements derived from the three-dimensional data were used to produce a series of discriminant functions for sex determination in the pooled sample, for which an accuracy of 77-80% was attained. Analysis of the calvaria and face, separately, has shown that the sex of damaged material can be diagnosed with a reasonable degree of accuracy (75-76%). The new functions for the pooled indigenous South African sample provide improved sex discrimination accuracy compared to those obtained by employing the commonly utilised statistics of Giles & Elliot (1963), even when a modified sectioning point is used. Functions calculated for the separate local populations gave variable and fairly low improvements in sexing accuracy. As the subdivisions at all levels are at present quite rapidly disappearing in South Africa, for most purposes it is now best to simply apply the pooled data functions for sexing crania. PMID- 15803768 TI - Predictive accuracy of sexing the mandible by ramus flexure. AB - Loth & Henneberg (Am J Phys Anthrop 99 (1996) 473) described a single morphological indicator of sexual dimorphism, namely the presence or absence of flexure on the posterior border of the mandible, with a predictive accuracy of 90.6-99.0%. In the other studies, which have criticized mandibular ramus flexure as sex indicator in adult and fossils specimens by the same method, the accuracy of sexing was found between 59.0% and 80.4% which is well below the reported 90.6 99.0% (Am J Phys Anthrop 107 (1998) 363; Am J Phys Anthrop 111 (2000) 573; Am J Phys Anthrop 111 (2000) 429; Homo 53 (2002) 97). It was indicated that the method sexed males more reliably than females and consistency was low. In the current blind test, 120 mandibles from forensic cases were examined for the presence or absence of mandibular ramus flexure. Virtually identical results were obtained when a second observer examined the same sample of mandibles. The results were then compared to records of the same cases resulting in 85.8% accuracy for the total sample. A marked difference was observed between sexes where the ramus shape was diagnostic for 92.6% of the males but for only 60.0% of the females (p < 0.001). When mandibles with excessive tooth loss (more than two molars missing) (n = 35) were removed from the sample (in normative sample), the overall predictive value increased to 90.6% (95.6% accuracy in males and 70.6% in females). Our results confirm that there are marked differences between the sexes in the predictive accuracy and this indicator is better in mandibles without molar tooth loss as mentioned by Loth & Henneberg. They also pointed out that the tooth loss is a potential source of error. On the other hand, in this study, there were no sexing errors when mandibles which ramus shape scores -1, 0 and +1 were extracted from the normative sample as the "sex indeterminate group". This study suggests that further assessment should be made only when both rami are available for observation, and the mandible shows either bilateral flexure (+2) or bilateral non-flexure (-2). The consideration of this new categorization can also be useful when using Loth & Henneberg's method. PMID- 15803769 TI - Secular changes in head dimensions of Japanese adult male students over eight decades. AB - The aim was to study secular changes in head dimensions of Japanese adult males. The subjects were all of Japanese birth and Japanese ancestry. The age range of the subjects was between 18 and 25 years. Four head measurements were taken: head length, head breadth, head height and head circumference. In addition, stature and body weight were also measured. All measurements were made by one observer (Fumio Ohtsuki) from 1998 to 2001. The present sample was compared with the series taken between 1910 and 1917 of Matsumura (J Fak Sci Imp Univ Tokyo Sec 1 (1925) 1) and the one measured in 1965 of Morita and Ohtsuki (Hum Biol 45 (1973) 151). The present study demonstrated the presence of larger means for head length, head breadth and cephalic index in the current sample than in their predecessors of about 35-85 years ago. Also, the present series displayed larger head circumference than that of the Morita and Ohtsuki series. Using ANOVA, head length and head breath showed significant (p<0.01) differences among all birth year cohorts from 1978 to 1983 of the current sample. Head height, head circumference and cephalic index did not display significant differences. However, the slope of the regression line indicated that all measurements as well as the cephalic index showed slightly decreasing tendencies during the investigated period. These results suggest that brachycephalization has been occurring for approximately about 35-85 years in adult Japanese males, but the change seems to have become reversed slightly during the period covered by the present sample. PMID- 15803771 TI - The people of Jabuticabeira II: reconstruction of the way of life in a Brazilian shellmound. AB - Sambaquis are huge shellmounds built along almost the entire Brazilian coast between 8000 and 600 years ago. In the present article, 14 osteological markers from 89 individuals excavated at the Sambaqui Jabuticabeira II (2890+/-55/2186+/ 60 BP) are analyzed in order to reconstruct the population's health status and way of life. The present palaeopathological findings (such as lower frequency of degenerative joint diseases in legs, as compared to arms, and the rarity of traumas) together with archaeological findings support the idea of nearby resource abundance and infrequent interpersonal competition. The presence of auditory exostoses mainly in males corroborates previous findings indicating the importance of marine resources. The low caries frequency and the high degrees of dental wear point to a diet poor in cariogenic food, and rich in abrasives such as sand, shell fragments and phytoliths. This suggests a broader diet, based on marine protein as well as plants, than previously thought. The etiology of cribra orbitalia could be explained by gastrointestinal parasites or other sources of physiological stress. These parasites, in turn, could have led to higher frequencies of infectious diseases, either by the debilitation of the immune system or by the direct contact with infectious agents. Despite the periods of illness various individuals experienced, the daily life among the builders of the Sambaqui Jabuticabeira II seems to have been relatively easy due to the abundance and predictability of resources and the paucity of violent traumas. PMID- 15803772 TI - Environmental stress in the Early Mediaeval Slavic population at Borovce (Slovakia). AB - Several palaeopathological indicators examined in skeletal samples are caused by stress during childhood and remain visible in adults. In this study, dental enamel hypoplasia was observed in 451 individuals from the Early Mediaeval (8th to beginning of 12th c. AD) Slavic skeletal series at Borovce (Slovakia). The presence of enamel hypoplasia was scored in all types of deciduous and permanent teeth. More than one-fourth (27.2%) of the individuals with preserved permanent teeth showed enamel hypoplasia. No significant differences in the occurrence of the enamel lesions were found between males and females. The age at development of hypoplasias was estimated for 74 individuals by measuring the distance of the defect from the cemento-enamel junction. The hypoplastic defects appeared most frequently between 2.5 and 3.0 years. Following the trends observed in the distribution of age at development of the enamel lesions between subadults and adults, individuals stressed earlier in life had a reduced ability to cope with later insults. High prevalence of enamel hypoplasia, especially among 10-14-year old growing juveniles, has led to the assumption that the Borovce population lived a considerably long period under conditions of high environmental pathogen load, and probably also suffered from some nutritional deficiencies. PMID- 15803770 TI - Pedal symphalangism in modern American and Japanese skeletons. AB - Pedal symphalangism is a surprisingly common heritable trait of the human foot. In individuals exhibiting the trait, the joint between the intermediate and distal phalanges of one or more lateral toes never develops, resulting in toes with two phalanges rather than three. This study was undertaken to explore variation in the frequency of pedal symphalangism among groups with widely different geographic ancestry, and to consider the applicability of this trait to skeletal biological distance studies. A total of 460 Euro-American, 191 African American and 99 Japanese skeletons were examined for presence of pedal symphalangism. The American individuals date to the first half of the 20th c, while the Japanese individuals date to the late 19th and early 20th c. Although the country of ancestry is unknown for most of the American individuals, the Euro Americans appear to be largely northern European, with roots in Germany, Ireland and Scandinavia, while the African-Americans are primarily descendants of slaves with roots in west African countries such as Nigeria, Benin, Togo, Ghana and Sierra Leone. Frequencies of fifth toe pedal symphalangism were calculated and compared among all three samples and found to be significantly higher in modern Japanese (83.7%) than in either Euro-Americans (46.4%) or African-Americans (44.0%). The Euro- and African-American frequencies were statistically indistinguishable for symphalangism of the fifth toe. In the fourth toe, however, the opposite result was found. The African-American frequency (7.9%) was significantly higher than the Euro-American frequency (2.6%), while no difference was found between the African-Americans and Japanese (11.7%). Since fourth toe pedal symphalangism has never been observed in the absence of fifth toe involvement, some of the same genes are clearly involved in producing the trait in both toes. However, differences in the pattern of fourth and fifth toe expression among the three groups identified in this study suggest that additional genetic or developmental factors may play a role in expression of pedal symphalangism in each of the toes. PMID- 15803774 TI - [New strategies against breast carcinoma. To outwit the resistance tendency of tumor cells]. PMID- 15803773 TI - [Breast carcinoma: new, much promising trends. The cancer anxiety of women -- will be taken better care soon?]. PMID- 15803775 TI - [Mammography is often not enough in high-risk patients. Tumor in image--screening with MRI?]. PMID- 15803776 TI - [Will the postoperative sessions in the cobalt bomb become the past soon? Radiotherapy has changed]. PMID- 15803777 TI - [Asthma overmortality under beta-2-mimetics. Does it depend on the drug itself or on false use]. PMID- 15803778 TI - [Adolescent with a chronic endocrinological disease. Are you fit to take over the patient?]. PMID- 15803779 TI - [Endocrinological transition clinic]. AB - Patients with chronic endocrinological diseases, whether congenital or manifesting in childhood, have to date been the domain of the pediatric clinic long beyond puberty into adulthood. The reason for this is the complete lack of adequate communication between pediatricians and internists specialized in endocrinology. However, intensive and constant cooperation between the two specialties is a prerequisite for the successful transition of adolescent to adult patients. The transition clinic is of central importance for the appropriate management and a stable status of the patient. Nevertheless, only very few university centers so far established such a clinic. Only by overcoming sectoral boundaries, and the creation of a network structure between university policlinic, non-university outpatient department, specialty practice and the family doctor can the intensity and quality of management be improved. The requirement fortransition clinics urgently needs to be met, also in the form of model projects. PMID- 15803780 TI - [The pediatric-to-adult transit clinic as a quality-securing measure in endocrinological diseases]. AB - The establishment of structured transit clinics for pediatric patients with endocrinological diseases entering adulthood is a prerequisite for ensuring the quality of care for patients with chronic endocrine diseases. Since such patients have been receiving constant special pediatric care on a large scale only during the last 20 years, and only now is an improvement in life expectancy and quality of life becoming evident for the individual diseases, a comprehensive store of experience, or even of evaluated data, is lacking. In order to preserve the quality achieved in pediatric care, transitional structures must initially utilize the experience and results achieved in the pediatric setting, and adapt them to the requirements of adult patients. This is an ongoing process, and additional resources are required for the transition, e.g. for joint medical counseling and care, as also for the continuing utilization of non-medical pediatric services. The basis for the implementation of such models is, in principle, provided by the Gesundheitsmodernisierungsgesetz, GMG (health care modernization act), and, for reasons of quality control, the necessary structures should be put in place as quickly as possible. PMID- 15803781 TI - [Differential diagnosis: what's behind the wart?]. PMID- 15803782 TI - [Genital piercings: if so, then by the physician. Pierced at a delicate spot]. PMID- 15803783 TI - [Treatment of allergic rhinitis: update 2005]. AB - Potential approaches to the treatment of allergic rhinitis are the avoidance of allergens and medication with chromone compounds, antihistaminics and glucocorticosteroids. The sole causally effective treatment is specific immunotherapy. Leukotriene receptor antagonists, anti-IgE antibodies and monoclonal CD-4-molecules, as also soluble cytokine receptors are potential therapeutic options, the value of which currently remains unknown. PMID- 15803784 TI - [Entirely without medical society and KV: colleagues built a successful care model]. PMID- 15803785 TI - [The role of incretins. Type 2 diabetes with new beginnings]. PMID- 15803786 TI - [Cardiovascular risk to be considered. Proinsulin test for stage-adjusted therapy]. PMID- 15803787 TI - [The 50th death anniversary of Albert Einstein (1879-1955). "The death is an old debt"]. PMID- 15803788 TI - [Look diagnosis. Green and blue. Without trauma]. PMID- 15803789 TI - [Predicting the aggressiveness of tumors]. PMID- 15803790 TI - [Artemisinin and successors]. PMID- 15803791 TI - [Paclitaxel for Alzheimer treatment]. PMID- 15803792 TI - [Drugs from nature. The biology of taxane]. PMID- 15803793 TI - [Assault on the mitotic spindle. The action mechanism of taxane]. PMID- 15803794 TI - [New microtubule stabilizers]. PMID- 15803795 TI - [ Development of new taxane formulations]. PMID- 15803796 TI - [Paclitaxel and docetaxel safe use: pharmacokinetics and potential interactions of taxanes]. PMID- 15803797 TI - [The action and toxicity of taxanes]. PMID- 15803798 TI - [Cost-benefit aspects of taxane therapy]. PMID- 15803799 TI - [Drug-eluting coronary stents]. PMID- 15803800 TI - [Administration of taxane-containing drugs]. PMID- 15803802 TI - [Is there danger of a bird flu pandemic?]. PMID- 15803803 TI - Lumbosacral neurenteric cyst with associated tethered cord in an adult. AB - Spinal neurenteric cysts are rare congenital lesions of endodermal origin that lead to spinal cord compression or tethering. These cysts are usually lined by mucus-secreting epithelium resembling that of the gastrointestinal tract. A 31 year-old man presented with low back and bilateral limb pain of three-year duration. Magnetic resonance imaging (MRI) revealed an intradural extramedullary heterogeneous mass lesion at L4 - L5 disc level causing severe compression and displacement of terminal portion of conus medullaris to the left side. Additionally, tethered cord, syringohydromyelia and spina bifida were present. He had also polysyndactyly on the left foot. The patient underwent laminoplasty and partial excision of the lesion through a posterior approach. Neurosurgical exploration revealed a large intradural, extramedullary solid lesion with cystic component, compressing the spinal cord. Histopathology confirmed a neurenteric cyst with mucin-producing cuboidal or columnar epithelium lining the cystic cavity. Clinical suspicion is important for an early diagnosis, and their early preplanned surgical removal should be the goal of treatment, which leads to a better outcome. This case is reported in view of rare congenital abnormality in an adult with peculiar presentation. PMID- 15803804 TI - Pituitary adenoma: a DNA flow cytometric study of 192 clinicopathologically characterized tumors. AB - A clinically, immunohistochemically and ultrastructurally characterized series of 192 pituitary adenomas was analyzed for DNA content by flow cytometry. Results were assessed not only relative to tumor immunotype, size, and invasiveness, but also with frequency of recurrence. Case selection was non-random; males predominated (1.8:1) and the ratio of macro-to-microadenomas was 4.2:1. Female patients were slightly younger and, in all adenoma categories, less often had invasive tumors: PRL (15%/30%), ACTH (17%/44%), LH/FSH (8%/27%) and null cell adenomas (0%/27%). With the exception of prolactin cell adenomas, similar proportions of macroadenomas and invasive tumors in all tumor subtypes were diploid and non-diploid. Prolactin adenomas differed in that tumors of males showed a high rate of non-diploidy (65%); such tumors were predominantly macroadenomas, but only 28% were invasive. Among GH-containing tumors 78% were macroadenomas, 40% were nondiploid, and the frequency of invasive macroadenomas was higher (49%) than in PRL tumors (21%). ACTH adenomas were mainly microadenomas (81%), their rate invasion (29%) and of non-diploidy being low (14%). Among "non-functioning" (LH/FSH, null cell adenomas), LH/FSH-producing tumors were all macroadenomas, but with low rates of invasion (23%) and non diploidy (9%). Null cell adenomas, nearly all macroadenomas, had similar low invasion rate (21%), but were more often non-diploid (39%). In all adenoma subgroups S-phase fractions were higher in non-diploid adenomas by an overall ratio of 2.1:1. Prolactin adenomas showed the highest (15.2%) and LH/FSH adenomas the lowest (5.6%) mean S-phase fraction. When compared to long-term follow-up, neither this parameter nor ploidy correlated with tumor size or invasiveness. Lastly, long-term follow-up showed ploidy to be an unreliable predictor of tumor persistence or recurrence. PMID- 15803805 TI - Cerebellar glioblastoma genetically defined as a secondary one. AB - We report here the case of a 29-year-old woman with cerebellar glioblastma. In the present case, tumor lesions were observed in each cerebellar hemisphere. The left-side lesion was diagnosed as glioblastoma, and the right-side lesion as malignant astrocytoma by histopathology. Immunohistochemistry revealed that the tumor cells of the left-side lesion was positive for p53, whereas epidermal growth factor receptors (EGFR) were negative in tumor cells from both sides. Genetic alterations were investigated using a genome DNA microarray (GenoSensor Array 300), which has led us to define this tumor as a secondary glioblastoma. The clinical presentation and genetic findings of this relatively rare entity are discussed. PMID- 15803806 TI - Loss of chromosome 13 in a case of soft tissue perineurioma. AB - Soft tissue perineuriomas are rare mesenchymal tumors that are derived from perineurial cells of the peripheral nerve sheath. Although the histological and immunohistochemical features of soft tissue perineuriomas are well described, little is known regarding the cytogenetic abnormalities in these tumors. Herein, we describe a case of a large (12.2 cm) soft tissue perineurioma that arose in the thigh of a 26-year-old Caucasian female. Histologically, the tumor was composed of a diffuse to fascicular arrangement of spindle cells with bland, elongated nuclei with long, thin, tapering cytoplasmic processes. The immunohistochemical profile was consistent with a perineurial cell origin with expression of epithelial membrane antigen, vimentin, and collagen type IV. Cytogenetic evaluation revealed loss of chromosome 13 as the sole abnormality in the majority of examined cells. In contrast to previous reports, we were unable to demonstrate deletion or structural abnormalities of chromosome 22 by either fluorescence in situ hybridization (FISH) or metaphase cytogenetics. This is the first report of loss of chromosome 13 in soft tissue perineurioma. Although never described in this group of neoplasms, loss of chromosome 13 has been identified in a large number of other soft tissue tumors, particularly sarcomas and malignant peripheral nerve sheath tumors. Herein, we discuss this case and provide a review of the literature. PMID- 15803807 TI - Clinical significance and neuropathology of primary MADD in C34-T and G468-T mutations of the AMPD1 gene. AB - OBJECTIVE: Primary myoadenylate deaminase deficiency (MADD) is probably the most frequent inborn metabolic myopathy with a prevalence of up to 2%. It is the result of mutations in the AMPDI gene, the most common of which is a C34-T transition in exon 2. The importance of the more rare mutation G468-T in exon 5 is uncertain. Primary objective was to elucidate the clinical significance of the enzyme disorder, which remains unclear since its first description in 1978. We further examined the existence of an association of MADD with other muscle disorders, such as malignant hyperthermia and rhabdomyolysis, as was suspected in earlier studies. MATERIAL AND METHODS: In a large collection of 1673 muscle biopsies that had been stored deep frozen we identified 33 cases of primary MADD, 12 of which without any other coinciding muscle diseases, by histochemical, biochemical and molecular genetic examinations. Clinical and laboratory data was collected. By additional examination of randomly chosen blood samples we identified one person carrying the rare compound heterozygosity C34-T/ G468-T, who was examined in clinical respects and a muscle biopsy was taken. RESULTS: As underlying mutation, the most common transition C34-T/C 143-T was detected in 33 cases. One patient carried the compound heterozygosity C34-T/G468-T. The overall frequency of MADD in the contingent was 1.8%. Only three patients out of 12 with isolated primary MADD suffered from muscle complaints, one of whom did not experience the typical symptoms of exercise related myalgia, muscle cramps and weakness as described by Fishbein. The patient carrying C34-T/G468-T was a fully healthy female. She had never experienced any muscle complaints. Any association with other neuromuscular disorders, if not completely ruled out, was found to be very unlikely. CONCLUSION: The results suggest that MADD itself is unlikely to be solely responsible for the manifestation of muscular symptoms. It is probable that either the loss of a compensation mechanism or coexistent disturbances in muscle metabolism which are unidentified so far are required for the emergence of complaints. PMID- 15803808 TI - Cerebral fat embolism: pulmonary contusion is a more important etiology than long bone fractures. AB - Lipid embolism is a serious and life-threatening problem and usually arises as a complication of severe trauma associated with long bone or pelvic fractures. It is generally thought that fat droplets enter the circulation at the site of fracture. In the systemic circulation, they become emboli to brain, kidney and other areas. Lipids are absorbed from the intestinal tract and transported into pulmonary tissue via thoracic duct and exposed to first catabolic procedures in the lungs. We have predicted that systemic lipid embolism may not occur unless bone fractures lead to pulmonary injury. This study was planned to investigate this hypothesis with respect to the role of pulmonary contusion and long bone fractures in the formation of cerebral fat embolism. Twenty male hybrid rabbits were included in this study. Pulmonary contusion was performed on half of the rabbits (n = 10) and femur fracture was applied to the remaining ones (n = 10). Ten days after procedure, all rabbits were sacrificed. Brain specimens were taken by frozen-section method and stained with Sudan black. Intraarteriolar lipid particles in the brain were examined microscopically. Cerebral fat embolism was detected in seven animals exposed to pulmonary contusion and only in one animal exposed to femur fracture. The mean number of branches of middle cerebral artery at midparietal level occluded with fat particles were higher in the pulmonary contusion group than in the long bone fracture group. In conclusion, we found that pulmonary contusion had more deleterious effects than long bone fracture in the formation of cerebral fat embolism. PMID- 15803809 TI - [The problem of teaching surgery in the third millennium]. PMID- 15803810 TI - [Revascularization of the carotid and vertebral arteries in the elderly]. AB - From January 1994 to July 2004, 323 patients underwent 348 revascularization of carotid bifurcation for atherosclerotic stenoses. Eighty eight patients (group A) were 75 year-old or older, whereas 235 (group B) were younger than 75 years. Postoperative mortality/neurologic morbidity rate was 1% in group A, and 1.4% in group B. At 5 years, patency and freedom from symptoms/stroke were, respectively, 91% and 92% in group A, and 89% and 91% in group B. None of these differences was statistically significant. In the same time period, 26 internal carotid arteries were revascularized in 24 patients, 75 or more aged, for a symptomatic kinking. Postoperative mortality/morbidity rate was absent, whereas, at 5 years, patency and freedom from symptoms/stroke were, respectively, 88% and 92%. Twelve vertebral arteries were revascularized in 12 patients, 75 or more aged, for invalidating symptoms of vertebrobasilar insufficiency. Postoperative mortality/neurologic morbidity rate was absent. In one case postoperative recurrence of symptoms occurred, despite a patent revascularization. Patency and freedom from symptoms/stroke were 84% and 75%, at 5 years. Revascularization of carotid and vertebral arteries in the elderly can be accomplished with good results, superposable to those of standard revascularization of carotid bifurcation in a younger patients' population. PMID- 15803811 TI - [Our experience in the surgical treatment of peritoneal hydatid disease]. AB - The Authors report on a series of 9 patients with peritoneal hydatidosis submitted to surgery during the period between 1974 and 2003. In 5 cases the cyst was solitary, while in 3 patients the cysts were multiple, in 1 spread and in 2 there was a concomitant liver hydatidosis. In 2 patients the disease was asymptomatic, while 4 cases were revealed by pain and 3 by an abdominal mass. The diagnosis was made at the operation in 3 patients, while it was performed by ultrasonography in 1 case, by CT scan in 4 and by both methods in 1. All patients were operated on: 7 underwent total cystectomy and partial cystectomy with external drainage of residual cavity was performed in one; the spread disease was managed by omentectomy, ovariectomy and appendectomy. Postoperative course was regular without mortality and major morbidity in all patients. Two recurrences occurred 71 and 20 months after removal of the primary cysts and they were managed by total cystectomy. The Authors believe that the diagnosis of peritoneal hydatid disease is today more accurate because of the new image techniques and the surgical procedure should be tailored to each patient depending on size, location and complications of the cyst, although the results of radical treatment are better. PMID- 15803812 TI - [Total splenectomy for a recurrent giant splenic cyst]. AB - Primary splenic cysts are a rare finding. Some are large and require surgical removal. The Authors report a case of a recurrent huge splenic cyst in a 41-year old female patient. A marsupialization was performed at another hospital 6 years before. Ultrasonography and computed tomography imaging revealed a cystic lesion in the spleen measuring approximately 20 cm in diameter. A total open splenectomy was performed. Postoperative course was uneventful. The histologic diagnosis was an epithelial cyst of the spleen with no atypical cells in the cyst wall, as previously found at the first operation. The epidermoid cysts have an epidermal lining, and prevention of recurrence is dependent on complete resection of the cyst wall preserving, whenever possible, the splenic tissue. Recurrence can be avoided with partial splenectomy in polar localization of the cyst, or complete removal of the cyst by "peeling" it off the splenic parenchyma. Marsupialization of the cyst, either via a laparoscopic or an open approach, is often ineffective. PMID- 15803813 TI - [Merkel cell tumor: a case report and literature review]. AB - Merkel cell tumor (MCT) is extremely rare, being discovered so far about 400 cases in literature. It is classified among neuroendocrine tumors. We report a case of MCT in the subclavicular region in a 93 years old woman. We confirm the efficacy of radiotherapy associated with octreotide, which these tumours express specific receptors for. PMID- 15803814 TI - [Appendiceal lithiasis: case report and review of literature]. AB - Appendiceal calculi are found in a limited percentage of patients. They are in association to an elevated prevalence of necrotic appendicitis and perforation of the appendix. The Authors report a case of acute appendicitis associated to appendiceal lithiasis with perforation of the organ. They emphasize the possibility to perform a preventive appendectomy in case of incidental appendiceal lithiasis. Besides they consider the differential diagnosis of the calcified images located in the lower abdominal quadrants with the aid of the various imaging methods available today, particularity sonography and computerized tomography. PMID- 15803815 TI - [Celiac artery aneurysm: urgent surgical case report]. AB - Visceral artery aneurysm account for up to 5-10% of overall artery aneurysm. Celiac artery aneurysm are reported in 4% of these patients. The most common etiologic findings are atherosclerosis and tunica media infective degeneration. The radiological imaging led to more accurate morphological and anatomical definitions of this pathology and improved elective and urgent surgical treatment of selected patients. The Authors report a case of celiac artery aneurysm. PMID- 15803816 TI - [A case of total spinal block during epidural anaesthesia]. AB - Most cases of total spinal block have been reported in the literature. The displacement of the catheter and the consequent dural perforation are the causes in large percentage of the patients (75%). The Authors describe this case for the importance of the causes and outcome of the patient. A 48 years old woman presented for hysterectomy for uterine fibromas. After having individualized the L3-L4 interspace, a test dose of 3 ml carbocaine 2% was injected. After that, the spinal block was obtained using ropivacaine 0.75% (total dose = 10 ml) injecting slowly, in following times, 5+5 ml of anaesthetic solution. The patient, perfectly conscious at first, presented a gradual increase of the difficulty in talking and breathing. Subsequently she showed a complete paralysis with loss of the consciousness, respiratory arrest, bilateral and symmetrical midriasis, as well as total areflexia. Endotracheal tube was placed. After eighty minutes from the end of the administration of the local anesthetic, spontaneous thoracic excursions appeared, even though of moderate ampleness, midriasis reduced. The patient recovered consciousness and sufficiently ventilated; therefore the endotracheal tube was removed. PMID- 15803817 TI - [A fecal fistula in patient who underwent an abdomino-perineal resection with a continent magnetic prosthesis: observations on an obsolete technique]. AB - The Authors consider a case of a patient who underwent an abdominal-perineal resection and presented a fecal fistula as a late complication of a magnetic prosthesis implant. After a revision of the literature, the Authors evaluate the reasons for abandoning this surgical technique of continence many years ago, underlying not only the complications observed during the experimentation but also the lack of those benefits for which this technique was proposed. PMID- 15803818 TI - Laparoscopic technique in enlarged cystic duct. AB - The risk of dislodgment of endoclips placed during laparoscopic cholecystectomy in enlarged cystic duct is higher with minor bile leak. From January 2000 to April 2004, we performed 1013 procedures; in 12 patients we have showed a enlarged duct ligated with 4 laparoscopic cholecystectomy endoclips after a 180 degrees rotation of the gallbladder during a retrograde cholecystectomy. We haven't registered complications and all the cases were discharged the first postoperative day. The method is safe and economically sound. PMID- 15803819 TI - Radical treatment of acute pilonidal abscess by marsupialization. AB - Simple drainage is the most common treatment of pilonidal abscess, but later definitive surgery is often necessary. However, radical treatment can also be performed in a single step by several procedures and even better results. The Authors describe the technique of marsupialization and analyse its long-term outcomes in a retrospective study. Between 1992 and 2001, 43 consecutive patients underwent drainage, curettage and marsupialization for acute pilonidal abscess. The procedure was carried out in outpatient setting (day surgery) after local or spinal anaesthesia. Management of the wound was left to the patient and was periodically checked at our out service. The average operating time was 20 minutes. Complete healing required 4-10 weeks in 95.3% of the patients; a persistent chronic fistula was observed in 4.7%. During the follow-up, 6 recurrences (14.6%) were observed in the healed patients. The treatment was then successful in 81.3% of the cases. Our experience shows that it is possible to carry out a radical management of pilonidal abscess in a single step with a high complete healing rate. Marsupialization proved to be a fast procedure and mainly feasible in the outpatient setting. PMID- 15803820 TI - Subclavian vein stenosis following totally implantable venous access device (TIVAD) implant by percutaneous approach. Case report. AB - Totally implantable venous access devices (TIVADs) are more and more used in oncologic patients. Early and late complications frequently occur with the techniques of implantation. Thrombosis is the most important and common late complication. On the opposite stenosis of the vessel is not frequently reported. The Authors report the case of a patient with subclavian vein stenosis after implantation of a TIVAD by percutaneous technique. On the basis of the published knowledges they try to explain this phenomenon and suggest an hypothesis for future works. PMID- 15803822 TI - [Atherothrombosis and acute coronary syndrome]. PMID- 15803821 TI - [Rare malignant tumors of the thyroid]. AB - The incidence of rare malignant tumors of the thyroid is about 4 cases/100,000 people and represent only 1.8% of all the thyroid cancers. When we talk about "rare" tumors, obviously, we do not refer to the most frequent cancers (papillary, follicular), or less frequent tumors (medullary, anaplastic), but to some types of thyroid tumors that have been almost always sporadically observed. Mucoepidermoid carcinoma and squamous carcinoma have been described in the literature. They present occasional papillary formation so that, according to some authors, could be considered as variants of the papillary carcinoma. Teratoma is another rare tumor which in the paediatric age is benign, but its prognosis could be unfavourable because it causes an important respiratory distress, while in the adult it presents a very aggressive clinical course like the anaplastic carcinoma. Lymphoma is the most frequent of the "rare" tumors of the thyroid (1-5% of all the thyroid cancers). It arises often in a setting of a long history of goitre and Hashimoto thyroiditis. Fine-needle aspirate is important to make diagnosis and to start a correct treatment which allows a 5 year survival up to 85% in the favourable cases. PMID- 15803823 TI - [Hyperhomocysteinemia as a clinical manifestation of thrombosis risk]. PMID- 15803824 TI - [Acute and chronic pancreatitis and pancreatic cancer: a chain of sequential events or independent diseases?]. PMID- 15803825 TI - [Blood vessel remodeling in chronic renal failure]. PMID- 15803826 TI - [Dynamics of corrected QT interval and heart rate in patients with early postinfarction angina]. AB - The subjects of the study were 135 patients with acute Q-wave and non-Q-wave myocardial infarction (MI) aged 36 to 78 years (mean 62.4 +/- 11.2 years). 59 patients (main group) developed early postinfarction angina (EPA). 76 patients (control group) had no stenocardia attacks within the entire period of hospital treatment. In the patients with EPA corrected QT interval dispersion was greater within the entire period of hospital treatment: 81.2 +/- 7.6 and 67.1 +/- 4.6 ms1/2 on the first day, 74.6 +/- 6.0 and 70.3 +/- 4.7 ms1/2 on the third day, 82.1 +/- 6.5 and 71.6 +/- 6.2 ms1/2 on the seventh day, 88.7 +/- 6.2 and 69.4 +/- 8.2 ms1/2 on the 28th day, respectively. Heart rate entropy decreased in both groups, but before discharge (mean 28th day) the patients with EPA demonstrated its significant reduction compared to the patients without EPA (3.89 +/- 0.16 and 4.13 +/- 0.09 bits, respectively, p < 0.05). Thus, recurrent myocardial ischemia in patients with MI, manifesting by EPA attacks, leads to the increase of QT interval dispersion and to heart rate rigidity which apparently mirrors greater probability of myocardial electric instability and increased risk of fatal arrhythmias in such patients. PMID- 15803827 TI - [Ferritin and other acute phase proteins in various forms of coronary heart disease]. AB - The purpose of the study was to determine clinical importance of high serum levels of ferritin, fibrinogen and C-reactive protein (CRP) in patients with various forms of coronary heart disease (CHD) such as stable angina, painless myocardial ischemia (PMI) and instable angina (IA). The subjects of the study were 60 patients with CHD, whose clinical variant (stable angina, PMI or IA) had been determined by stress echocardiography. The control group consisted of 20 patients, not suffering from CHD, but having cardiovascular risk factors (arterial hypertension, dyslipoproteinemia, male gender, obesity, elderly age). All patients underwent routine clinical examination and biochemical blood tests. Serum levels of CRP, fibrinogen and ferritin were highest in the patients with IA and significantly differed from those in the control group. The difference in serum iron levels and total iron-binding capacity in serum (TIBC) between the groups were insignificant. Correlations between serum level of iron, TIBC and ferritin level were found neither in CHD patients (r = 0.1) nor in the control group (r = 0.15). No correlation between serum level of ferritin and CRP level was observed in the control group, but in all CHD groups this correlation was significant. The strongest correlation between these values was observed in the patients with IA. Besides, correlations between serum levels of ferritin and CRP (r = 0.46, p < 0.02) and between ferritin and fibrinogen levels (r = 0.39, p < 0.05) were found in the patients with IA. In patients with CHD, especially those who have IA, serum ferritin should be considered among acute phase proteins, reflecting destabilization of atherosclerotic plaque. PMID- 15803828 TI - [Characteristics of the pathogenesis, manifestations and clinical course of psychogenically induced myocardial infarctions]. AB - With the purpose of investigating peculiarities of psychogenically induced myocardial infarction (PIMI) 82 patients with primary myocardial infarction (MI) were chosen as the subjects of the given controlled study and divided into two groups. The main group consisted of 33 patients, the rest 49 formed the control group. The study showed that coronary atherosclerosis was more pronounced in the patients of the main group, among whom cases of exertional angina in past history were more frequent, and who had more pronounced coronary calcinosis compared to the patients of the control group. At the same time, the clinical course of MI in such patients is relatively benign, but it is more often complicated by early postinfarction angina. All this suggests that the pathogenesis of PIMI differs from that of "classic" MI. In particular, PIMI may be associated with the involvement of more distant parts of coronary vessels. Patients with PIMI seem to need to be regarded as having high risk of repeated coronary disasters. PMID- 15803829 TI - [Laser correction of microcirculation disorders in patients having CHD with hypercholesterinemia]. AB - The study demonstrates that hypercholesterinemia in patients with coronary heart disease (CHD) is associated with functional depression of microcirculation, increase in total peripheral vascular resistance, reduction in the functional efficiency of heart and decrease in activity tolerance. After receiving a course of low-intensity infrared laser radiation treatment the patients displayed positive changes in blood lipid spectrum, which was associated with improvement in microcirculation, decrease in afterload, increase in economization of heart functioning and activity tolerance. The obtained results demonstrate that the hypolipidemic effect of laser radiation is a substantial factor in the regression of CHD manifestations. PMID- 15803830 TI - [Effect of amlodipine on intravascular thrombocyte activity in patients with arterial hypertension and metabolic syndrome]. AB - In order to evaluate therapeutic effects of amlodipine on intravascular thrombocyte activity (ITA) in patients having arterial hypertension (AH) with metabolic syndrome (MS), 20 patients were treated with amlodipine for 1 month. The researchers monitored changes in anthropometric indices, blood lipid spectrum, lipid peroxidation in serum and thrombocytes, anti-oxidant protectability of platelets and the liquid part of blood and ITA. Amlodipine application in patients having AH with MS reduced peroxidation syndrome and optimized thrombocyte aggregation. Long-term administration of amlodipine stabilizes the achieved effect. To lower body weight in patients having AH with MS administration of amlodipine should be combined with use of non-drug means. PMID- 15803831 TI - [Hormone-producing function of the ovaries in young and middle-aged women with arterial hypertension and coronary heart disease]. AB - The study demonstrates that arterial hypertension itself, judging by main anthropometric variables such as body weight index and waist to hip ratio, together with the estrogen-producing function of the ovaries, evaluated by measuring follicle-stimulating hormone levels, contributes substantially to the forming of abdominal obesity in normo- and hypertensive women. All the above confirms close correlation between hemodynamic and metabolic disturbances which eventually result in coronary heart disease in young and middle-aged women. PMID- 15803832 TI - [Specifics of lipid exchange in patients with atherosclerotic lesions of the coronary and cerebral arteries]. AB - Atherogenic shift in blood lipid spectrum, which may manifest by the increase of serum levels of cholesterin, low-density lipoproteins (LDL), triglycerides, very low-density proteins (VLDL) or the decrease of serum levels of antiatherogenic high-density lipoproteins (HDL), is one of the leading risk factors of atherosclerosis. Patients with atherosclerosis often have normal or subnormal level of cholesterin together with hypertriglyceridemia or no hyperlipidemia (HL) together with decreased serum level of HDL. Among coronary atherosclerosis (CorA), cerebral atherosclerosis (CA) and multifocal atherosclerosis the first condition has been explored more than the others, whose ethiopathogenic features has not been sufficiently studied yet. However, sufficient knowledge of ethiopathogenic features of atherosclerotic process in various blood vessel basins would allow revealing the so called predictors of multifocal lesions and carrying out their timely prophylaxis or inhibiting their progression. Investigation of lipid exchange showed that the most atherogenic alterations are characteristic of CorA and multifocal atherosclerosis. CA is characterized chiefly by HL type IV, while HP type II is more typical of CorA and multifocal atherosclerosis. Triglycerides have been demonstrated to influence mostly the progress of CA while LDL chiefly promote the dissemination of CorA. PMID- 15803833 TI - [The condition of intestinal microflora in patients with hematologic tumors receiving polychemotherapy]. AB - The subjects of the study (105 patients with blood system tumors), were divided into 4 groups. The patients of the first group received only cytostatic therapy. The subjects of the second, third and fourth groups were administered a combination of cytostatic and antibacterial agents. In addition to this, the patients of the third and fourth groups received prebiotic preparations as means of corrective therapy. The results obtained by measurement of short-chain fatty acids (SCFA) in feces by means of liquid-gas chromatography were used as criteria for evaluation of the therapy influence on intestinal microflora. The study shows that the cytostatic and combined treatment worsen the disbalance between aerobe and anaerobe populations, which manifests in SCFA spectrum alteration. Use of prebiotic preparations results in improvement of intestinal microflora condition or prevents the aggravation of its disturbance caused by the treatment of the fundamental illness, which becomes apparent in stabilization or normalization of SCFA profile in feces. Study of SCFA makes it possible to monitor microbiocenosis, adjust corrective therapy individually and evaluate its effectiveness. PMID- 15803834 TI - [Metabolic aspects of the prediction of outcome in chronic viral hepatitis]. AB - The researchers evaluated the condition of lipid peroxidation and antioxidant protection system together with biochemical, immunological and morphological values in 117 patients with chronic hepatitis (CH) in virus replication phase (28 patient had CH type B, 66--CH type C and 28--CH type B+C), and estimated the importance of their interrelations to the prediction of disease outcomes. PMID- 15803835 TI - [Diagnosis and treatment of posttraumatic heart diseases]. PMID- 15803836 TI - [Difficulties in the diagnosis and successful treatment of pheochromocytoma]. PMID- 15803837 TI - [Current view of some obscure points regarding peptic ulcer and Helicobacter pylori invasion]. PMID- 15803838 TI - [The role of annual conferences held in Smolensk in the development of gastroenterology in our country]. PMID- 15803839 TI - [The 140th anniversary of the Department of Propaedeutics of Internal Diseases at the I. M. Sechenov Medical Academy in Moscow]. PMID- 15803840 TI - [Between physiologic aging and dysfunction. How much testosterone does a man need?]. PMID- 15803841 TI - [Patient requests testosterone. When is it actually indicated? (interview by Dr. Ramona Volkert)]. PMID- 15803842 TI - [When physicians become ill. Do not accept special treatment!]. PMID- 15803843 TI - [Suspected Crohn disease diagnosis: endoscopy alone is not reliable]. PMID- 15803844 TI - [Coincidence of depression and cardiovascular diseases. Heart pain has 2 faces]. PMID- 15803845 TI - [The immunosuppressed patient with opportunistic infection. This is now the responsibility of the general practitioner]. PMID- 15803846 TI - [Myoblasts from the leg into the heart: can infarct damage be rectified in this manner? (interview by Dr. Judith Neumaier)]. PMID- 15803847 TI - [We can no longer ignore the fact that COPD will soon be the third most common cause of death]. PMID- 15803848 TI - [Early diagnosis of chronic obstructive pulmonary disease (COPD)]. AB - Since the early detection of COPD is problematic, nonobstructed smokers with a chronic productive cough are initially assigned to the COPD risk group o. Although there is still a lack of evidence that early pharmacological intervention is associated with benefits in terms of disease progression, the earliest possible diagnosis is still considered a desirable goal. For the sooner triggering noxae, such as cigarette smoke, are eliminated, the more positive are the effects on the subsequent course of the illness. When establishing the diagnosis, a careful case history is of particular importance. With the aid of various diagnostic pulmonary function tests, degrees of severity can be differentiated and the course of COPD can be determined. Since the end of 2004, structured therapeutic programs for COPD have become available. PMID- 15803849 TI - [COPD in the year 2005]. AB - COPD is a widespread illness with an increasing prevalence and mortality rate. Current knowledge of its pathophysiology has it that COPD is not simply limited to the lungs, but must be understood as a systemic inflammatory reaction. The ATS/ERS guideline goes beyond the recommendations of the GOLD guideline, and takes account of the peculiarities of the European and North American healthcare systems (www.ersnet.org). In this context, the role of inhaled steroids, which is currently still under lively discussion, has to be revisited. Additional possible approaches include the administration of appetite stimulants and dietary measures, and the application of iron or erythropoietin. The TORCH study- currently the largest ongoing COPD study--is investigating the influence of inhaled glucocorticoids and long-acting beta2 agonists on mortality. PMID- 15803850 TI - [Modern inhalation therapy]. AB - Treatment with aerosols is the major form of treatment of obstructive airway disease. It has become possible not only as a result of improved medication, but also, and in particular, the continuing technological development of inhalation systems. The large selection of widely differing systems increases the chance of achieving effective treatment through appropriate individual application and good compliance. Even modern inhalation systems are not ideal in every respect and for every patient and disease stage. Only when correctly applied are almost all currently available systems clinically effective. When deciding what constitutes the best option, technological aspects of the systems together with the resulting demands made on the patient, as well as the mental and manual abilities and ventilatory characteristics of the patient, must be taken into account. PMID- 15803851 TI - [Olfactory and gustatory disorders--causes, diagnosis and treatment]. AB - Olfactory and gustatory disorders are by no means uncommon, and may be associated with an appreciable impairment of the patient's quality of life. In many cases, diagnosis and treatment is not easy, and necessitates interdisciplinary cooperation between the general practitioner, internist, ENT specialist, neurologist and psychiatrist. Many of the non-evidence-based treatments still applied in hospitals and the physician's office should, for reasons of quality control and to avoid polypragmasy possibly associated with undesirable side effects, be employed with reservations. With reference to the guidelines issued by the working group Olfactologie/Gustologie der Deutschen Gesellschaft fur HNO Heilkunde, Kopf- und Hals-Chirurgie (Olfactology/Gustology of the German Society of ENT Medicine, Head and Neck Surgery), a review of the causes, diagnosis and treatment of olfactory and gustatory disorders [3,4] is presented. PMID- 15803852 TI - [Reorganization of dated paradigms here. Essential hypertension is out]. PMID- 15803853 TI - [Increasinge general practice break-ins. Do not give insurance grounds for inadequate restitution]. PMID- 15803854 TI - [Incidental hypo- or hypercalcemia finding. Parathyroid hormone is the key to diagnosis]. PMID- 15803855 TI - [Diagnosis at a glance. Measles, rubella or scarlet fever?]. PMID- 15803857 TI - Molecular specificity of 5-androstenediol as a systemic radioprotectant in mice. AB - We compared in vivo radioprotective efficacy of 5-androstenediol (5-AED) to that of ten other steroids: 17alpha-androstenediol, dehydroepiandrosterone, 5 androstenetriol (AET), 4-androstenedione (AND), testosterone, estradiol, fluasterone, 16alpha-bromoepiandrosterone, 16alpha-fluoro-androst-5-en-17alpha-ol (alpha-fluorohydrin, AFH), and 16alpha-fluoro-androst-5-en-17beta-ol (beta fluorohydrin). Steroids were administered 24 or 48 hr before, or 1 hr after, whole-body gamma-irradiation. Two days after irradiation at 3 Gy, blood elements were counted. In addition, after irradiation at 9-12.5 Gy, survival was recorded for 30 days. The results showed radioprotective efficacy was specific for 5-AED. One other steroid, AFH, demonstrated appreciable survival effects but was less efficacious than 5-AED. AND and AET produced slight enhancement of survival in some experiments. This is the first demonstration that the prophylactic window for survival enhancement by 1 subcutaneous (s.c.) injection of 5-AED is as long as 48 hr in mice. Moreover, the results indicate that 1 s.c. injection of 5-AED 1 hr after irradiation is much less effective than 1 injection 24-48 hr before irradiation. Comparing the molecular features of steroids with radioprotective efficacy leads to the following conclusions: 1) these effects are due to interaction with specific receptors, since s.c. injection of extremely similar molecules with the same physicochemical properties as 5-AED were not radioprotective; 2) the 17-hydroxyl group is essential; 3) this group must be in the beta configuration in the absence of nearby side groups; 4) a halogen atom at 16 changes the 17-hydroxyl specificity to alpha; 5) the 3beta-hydroxyl group is not essential; 6) addition of a 7beta-hydroxyl group is deleterious; and 7) the effects are not due to activation of sex steroid receptors. PMID- 15803858 TI - Dendroaspis natriuretic peptide induces the apoptosis of cardiac muscle cells. AB - Early heart failure is characterized by elevated plasma Dendroaspis natriuretic peptide-like immunoreactivity (DNP-LI). However, the direct effects of DNP on heart or the heart-associated cell system are not well known. Therefore, we investigated whether DNP induces the apoptosis of H9c2 cardiac muscle cells. H9c2 cardiac muscle cells and rat neonatal cardiomyocytes were treated with various concentrations of DNP. Cell viability and nuclear morphology change were determined by trypan blue staining and Hoechst 33258 staining, respectively. Caspase-3-like activity was measured using specific fluorogenic substrates. Pro and antiapoptotic proteins were assayed by Western blotting. DNP induced the apoptosis of H9c2 cardiac muscle cells in a dose-dependent manner. Maximum effects occurred at 100 nM concentration of DNP, with a 7-8-fold increase in apoptotic cells, to reach a maximum apoptotic index of 17%. We also identified that H9c2 cardiac muscle cells expressed Natriuretic peptide reactor -A and -B, which respond to DNP to generate cGMP. The treatment with DNP also markedly reduced levels of Bcl-2, inhibitor of apoptosis protein-1, and inhibitor of apoptosis protein-2 and increased the level of Bax and cytochrome c release into cytoplasm and subsequent caspase-3 activation, which co-occurred with increased apoptosis. DNP-induced apoptosis was mediated by cyclic GMP, and this effect was mimicked by dibutylyl-cGMP (30 microM), a membrane permeable analog of cGMP. Furthermore, DNP-induced apoptosis was observed in rat neonatal cardiomyocytes. These results suggest that DNP induces the apoptosis of H9c2 cardiac muscle cells and of cardiomyocytes via cGMP and demonstrate that the operative mechanism includes the regulation of Bcl-2 family proteins. PMID- 15803856 TI - Effect of Tinospora cordifolia on the antitumor activity of tumor-associated macrophages-derived dendritic cells. AB - We and others previously have reported that extract prepared from medicinal plant Tinospora cordifolia shows a wide spectrum of immunoaugmentary effects. Tinospora cordifolia was shown to upregulate antitumor activity of tumor-associated macrophages (TAM). In this article we present evidence to show that an alcoholic extract of Tinospora cordifolia (ALTC) enhances the differentiation of TAM to dendritic cells (DC) in response to granulocyte/macrophage-colony-stimulating factor, interleukin-4, and tumor necrosis factor. DC differentiated in vitro from TAM that were harvested from tumor-bearing mice after i.p. administration of ALTC (200 mg/kg body weight) 2 days posttumor transplantation shows an enhanced tumor cytotoxicity and production of tumoricidal soluble molecules like TNF, IL-1, and NO. Adoptive transfer of these TAM-derived DC to Dalton's lymphoma-bearing mice resulted in prolongation of survival of tumor-bearing mice. This is the first report regarding the differentiation and antitumor functions of TAM-derived DC obtained from tumor-bearing host administered with ALTC. The possible mechanisms involved also are discussed. PMID- 15803859 TI - Triptolide attenuates endotoxin- and staphylococcal exotoxin-induced T-cell proliferation and production of cytokines and chemokines. AB - Proinflammatory cytokines mediate the toxic effects of superantigenic staphylococcal exotoxins (SE) and bacterial lipopolysaccharide (LPS). Triptolide, an oxygenated diterpene derived from a traditional Chinese medicinal herb, Tripterygium wilfordii, inhibited SE-stimulated T-cell proliferation (by 98%) and expression of interleukin 1beta, interleukin 6, tumor necrosis factor, gamma interferon, monocyte chemotactic protein 1, macrophage inflammatory protein (MIP) 1alpha, and MIP-1beta by human peripheral blood mononuclear cells (PBMC). It also blocked the production of these cytokines and chemokines by LPS-stimulated PBMC in a dose-dependent manner. These results suggest that triptolide has potent immunosuppressive effects even counteracting the effects of superantigens and LPS. It also may be therapeutically useful for mitigating the pathogenic effects of these microbial products by downregulating the signaling pathways activated by both bacterial exotoxins and endotoxins. PMID- 15803861 TI - Severe cholestatic hepatitis caused by azathioprine. AB - A male patient receiving azathioprine treatment for discoid lupus erythematodes developed severe cholestatic hepatitis between 14 and 21 days after initiation of the treatment with peak bilirubin levels of 62.4 mg/dL. Other causes of hepatic dysfunction including viral hepatitis were clinically and serologically excluded. Liver biopsy revealed cholestatic hepatocellular damage. At 14 days after discontinuation of azathioprine the liver function (transaminases and bilirubin) began to improve. Only alcaline phosphatase and gamma-glutamyl transferase remained elevated even after 4 months. This case argues for an idiosyncratic cholestatic hepatocellular damage caused by azathioprine. PMID- 15803860 TI - Study of innervation, sensory neuropeptides, and serotonin in murine contact allergic skin. AB - Density of nerve fibers, axonal growth, calcitonin gene-related peptide (CGRP), and substance P, and serotonin immunoreactivity as well as concentration were all determined in a murine model of contact allergy. Female Balb/c mice were sensitized on the back with oxazolone and 6 days later challenged with the same antigen on the dorsal surface of the ears, while control mice received the vehicle only. Then, 24 hr postchallenge, one ear was processed for immunohistochemical staining, while the other was frozen and processed for gas chromatography-mass spectrometry or radioimmunoassay (RIA). Protein gene product 9.5 (PGP 9.5) positive nerve fibers showed a tendency to increase in inflamed ears versus control ears in epidermis as well as the dermis. Growth-associated protein-43 (GAP-43) positive fibers in the epidermis were increased (p < .01) in inflamed ears, compared with control ears, as was the case for the dermal fibers, indicating increased axonal growth. Total (epidermis and dermis) numbers of CGRP and substance P positive nerve fibers tended to increase in the inflamed skin in contrast to control skin. In contrast, RIA demonstrated a lower (p < .05) concentration of CGRP in the inflamed ears compared with controls and a tendency for substance P to decrease in concentration in eczematous ears versus controls. There was no difference in serotonin concentration, or in the number of serotonin positive mast cells, between the inflamed and control skin, whereas semiquantification of serotonin positive platelets showed an increase in the inflamed (+/+) compared with control ears (+). Our results indicate that 24 hr after being challenged with the antigen, at the peak of murine skin inflammation, axonal growth, sensory neuropeptides, as well as serotonin may be involved. PMID- 15803862 TI - Distinct cytokine release profiles from human endothelial and THP-1 macrophage like cells exposed to different amphotericin B formulations. AB - Amphotericin B(AmB) formulations, Fungizone, and Amphotec caused substantially greater proinflammatory cytokine release than AmBisome (L-AMB) and Abelcet in TPA differentiated THP-1 macrophages as determined by antibody based protein arrays. Lipopolysaccharide but not AmB induced significant pro-inflammatory cytokines in human endothelial cells. PMID- 15803863 TI - Thymidine phosphorylase profiles in nonmalignant and malignant pancreatic tissue. Potential therapeutic role of capecitabine on tumoral and endothelial cells and tumor-infiltrating macrophages. AB - The drug capecitabine (CAP) is a thymidine Pi-deoxyribosyltransferase (TP) activated oral fluorpyrimidine that generates 5-fluorouracil (5-FU), preferentially, within tumors. Here, in 38 patients with pancreatic cancer we analyzed immunohistochemical TP expression in pancreatic cancer tissue (PCT) and adjacent nonmalignant pancreatic tissue (ANMPT). In addition, a correlation with the main clinical pathological features was made. Furthermore, TP-positive macrophages (MO) isolated from neoplastic tissue were determined. The mean of TP positive epithelial cells and endothelial cells in terms of microvessel density was significantly higher in PCT than in ANMPT. Because pancreatic cancer is sensitive to 5-FU, TP-activated oral CAP in tumoral and endothelial cells and tumor infiltrating MO could increase the concentration of 5-FU at tumor site, thus resulting in an enhanced antitumor activity. PMID- 15803864 TI - Effects of antioxidants coenzyme Q10 and lipoic acid on interleukin-1 beta mediated inhibition of glucose-stimulated insulin release from cultured mouse pancreatic islets. AB - During the development of the autoimmune disease, insulin-dependent diabetes mellitus (IDDM) islet cell death is thought to be mediated in part by oxygen and nitrogen free radicals and interleukin 1beta (IL-1beta), secreted by activated macrophages. Free radicals disrupt the homeostasis of biological systems by damaging major constituent molecules such as lipids, proteins, and DNA. Islet cells are quite susceptible to oxidative damage due to low levels of antioxidant enzymes involved in free radical consumption. If IDDM is associated with an imbalance of oxidative stresses and antioxidant responses in islet cells, then it may be possible to ameliorate disease by supplementating antioxidant defenses. In this study, the antioxidants coenzyme Q10 and lipoic acid were able to block IL 1beta-mediated inhibition of glucose-stimulated insulin secretion from islet cells at 10(-12) M and 10(-9) M, respectively. PMID- 15803866 TI - Bone marrow progenitor cells from chemically exposed workers display an intrinsic ability for autonomous proliferation. AB - In this study, the autonomous proliferation of bone marrow progenitor cells (CFU C), a pathological phenomenon observed in many hematological abnormalities, was investigated in 31 individuals who had been diagnosed as having neutropenia. Of these subjects, 18 had been chronically exposed (range of exposure 5-30 years) to a variety of petroleum distillates. We observed that the group of exposed individuals presented higher numbers of autonomous CFU-C when compared with those unexposed subjects. In addition, follow-up data demonstrated that 20% of the exposed population (4 of the 18) developed malignant hematological diseases. The autonomous CFU-C obtained from all individuals studied was composed predominantly of macrophages. This suggests an involvement of these cells in the development of hematological abnormalities, probably as a result of increased production of chemical myelotoxic metabolites. PMID- 15803865 TI - Mercury alters endotoxin-induced inflammatory cytokine expression in liver: differential roles of p38 and extracellular signal-regulated mitogen-activated protein kinases. AB - Mercury is a widespread metal in the environment and consequently large populations are currently exposed to low levels of mercury. Endotoxin, a component of the Gram-negative bacteria, promotes inflammatory responses. We recently reported that mercury modulates the production of nitric oxide and various inflammatory cytokines induced by endotoxin in a macrophage cell line (Nitric Oxide 2002, 7:67). The present study was designed to determine the impact of mercury on endotoxin-induced inflammatory cytokine expression and corresponding signal transduction in mouse liver. Male BALB/c mice were exposed continuously to 0, 0.3, 1.5, 7.5, or 37.5 ppm of mercury in drinking water for 14 days and at the end of the treatment period lipopolysaccharide (LPS, 0.5 mg/kg) was injected intraperitoneally 2 hr prior to euthanasia. The doses of mercury and LPS did not cause hepatotoxicity as indicated by unaltered circulating alanine aminotransferase and aspartate aminotransferase levels. Mercury decreased liver glutathione (GSH) and with LPS additively decreased GSH. Mercury activated p38 mitogen-activated protein kinase (MAPK) and additively increased LPS-induced p38 MAPK phosphorylation. In contrast, mercury alone had no effect on activation of extracellular signal-regulated kinase (ERK) but inhibited LPS-induced ERK activation. Mercury increased the expression of tumor necrosis factor alpha (TNFalpha) and further potentiated LPS-induced TNFalpha expression. Mercury did not affect LPS-induced interleukin (IL)-1beta expression but decreased LPS induced IL-6 expression. Results indicated that low levels of mercury augment LPS induced TNFalpha expression by altering GSH and p38 MAPK. Mercury modulates LPS induced p38 and ERK activation and downstream TNFalpha and IL-6 expression in mouse liver. PMID- 15803867 TI - Lymphoreticular system in fish: erythrocyte-mediated immunomodulation of macrophages contributes to the formation of melanomacrophage centers. AB - In fish, melanomacrophage centers (MMC) represent the equivalent of germinal centers of birds and mammals. They consist of aggregates of macrophages filled with fragments of cells, mainly erythrocytes, and various pigments such as hemosiderin, lipofuscin, and melanin. The kidney, the spleen, and the liver contain many MMC either moving freely or encapsulated in the context of the lymphoreticular tissue. Here, we have demonstrated for the first time that MMC exist in the kidney, spleen, and liver of the rainbow trout Salmo gairdneri Richardson by using traditional stainings and cytoenzymatic methods: periodic acid-shiff, alpha-naphthyl acetate esterase, and peroxidase. Finally, in the view of the already described peripheral interaction between fish erythrocytes (FE) and macrophages (MO), we postulate that these cellular aggregates (rosettes) travel to distant organs, thus contributing to the formation of MMC in the kidney, spleen, and liver. PMID- 15803868 TI - Effect of formaldehyde gas exposure in a murine allergic contact hypersensitivity model. AB - To clarify the effect of formaldehyde (FA) gas exposure on contact hypersensitivity (CHS), CHS reactions against 2,4,6-trinitrochlorobenzene (TNCB) was studied in BALB/c mice with a low dose of FA gas exposure. The TNCB-induced CHS reactions were slightly suppressed by the FA gas exposure immediately after sensitization, whereas they were significantly enhanced and prolonged in mice continuously exposed to FA gas before and after sensitization. We showed that exposure to FA gas enhanced the Th2 dominant responses in draining lymph node (LN) in early stage of CHS. In contrast, T cell subsets and their intracellular cytokine production in the draining LN were similar during the early stage of CHS by FA gas exposure during the sensitization phase. The percentage of CD8+ T cells was increased, and the percentage of CD4+CD25+ T cells was decreased in the FA gas-exposed group at 72 hr after elicitation. These results indicate that FA gas exposed might influence regulatory T cells. Furthermore, in the chronic CHS model that was repetitively elicited with TNCB, more intensive and prolonged CHS reactions, and increased numbers of mast cells were found in the FA gas-exposed group at 4 hr after elicitation than in the control group, FA gas exposure may alter the intensity of allergic CHS. PMID- 15803869 TI - Long-term toxic effects of radiations: sarcomatoid carcinoma and multiple basal cell carcinoma of the limbs in chronic radiodermatitis. AB - A 73-year-old woman was referred to our attention for the presence of severe chronic lymphoedema and neoplastic lesions of the limbs. When she was 19 years old, the patient had undergone X-ray epilation of the limbs to remove unwanted body hair. The neoplastic lesions, observed 2 years before our observation, were excised, and histological examination revealed multiple basal cell carcinomas and a spindle squamous cell carcinoma on the left thigh. The case we observed offers some interesting points of consideration as it shows that in nonexposed areas cutaneous carcinomas can appear many years after irradiation and that the same treatment consequently requires careful follow-up for life. Another point of interest is the particular hystological aspect of neoplastic lesions in the site of radiodermatitis that requires in-depth immunohistochemical investigation for an exact diagnosis. Patients with previous treatment with ionizing radiation also in nonexposed areas need lifelong follow-up to identify cutaneous tumors at an early treatable stage. PMID- 15803870 TI - Saliva secretory IgA antibodies against molds and mycotoxins in patients exposed to toxigenic fungi. PMID- 15803871 TI - [Issue in honor of Professor Gyula Sebestyen, 1887-1954]. PMID- 15803872 TI - [50th anniversary of the death of Professor Gyula Sebestyen (1887-1954)]. PMID- 15803873 TI - [Surgery of the thymus]. AB - On the occasion of the 50th anniversary of Gyula Sebesteny's death, an overview was prepared on the surgery of the thymic gland that was closely connected to his thoracic surgical activity. His results with thymectomy in myasthenia gravis were published as the second study on this topic in Europe and the first in Hungary. The early thymic surgery was based on pathological observations. Myasthenia gravis, a well-known disease for long, occurred frequently with pathological lesions of the thymus. Autopsies suggested potential causative relationship. The early period of thymic surgery was characterised by sporadic interventions and high mortality. The new period of the thymic surgery started with the American Blalock's 20 operations, and accomplished by the operative activity of Keynes including 281 thymectomies. They were followed by many European, American and Asian surgeons. Even today, there are many hotly debated topics, like indication and type of operation, prognostic factors, perioperative care, diagnosis, treatment and pathology of the thymic tumor. With fine-tuning the indication, the development of operation techniques, neurology, anaesthesiology and intensive care, imaging techniques and oncology and with introduction of new drugs the early and late results are improving. Thanks to Dr. Sebesteny, Hungarian thoracic surgeons joined early the international thoracic surgical activities. Currently in Hungary there are two major centers for myasthenia gravis surgery, and all important thoracic surgical departments in the country treat thymic tumor cases. PMID- 15803874 TI - [Our experience in the surgical treatment of tracheal diseases]. AB - A case control study is presented reviewing the outcomes of 32 consecutive major tracheal surgeries. The audit is based on our six and half year experience. MATERIAL AND METHODS: There were 11 (neo)esophageal communications, 9 direct injuries (iatrogenic and trauma), 8 endotracheal obstructions and 4 inflammatory processes requiring surgical intervention. Six sutures of the trachea, 13 resections with anastomosis, 2 external fixations and 11 combined esophagotracheal procedures were performed. RESULTS: The 30-day mortality was 12.5% (4/32) with a dominance of the inflammatory cases. The overall mortality was 21.8% due to the late complications of the surgeries for esophagotracheal sinuses. There was no death. CONCLUSIONS: Independent factors influencing outcome seemed to be: underlying pathology of the trachea, timing of intervention, nutritional state of the patient, and surgical technique. Tracheal replacement can be considered the key element of further process in this field of thoracic surgery. PMID- 15803875 TI - [Results and alternatives in the surgical treatment of lung cancer]. AB - OBJECTIVE: The aim of our study was to analyse the results of surgical treatment in lung cancer cases. PATIENTS AND METHODS: Between 1990 and 1999, 860 patients were operated on for primary lung cancer. In 775 patients (90.1%), resection was performed, 85 patients underwent explorative thoracotomy. RESULTS: The 5-year survival rate was 45.3%, the mean survival time was 47 months. In the univariate analysis, the radicality of the resection (p < 0.0001), the pT stage (p < 0.0001), the pN stage (p < 0.0001) and the histological type (p = 0.0039) had a significant correlation with the survival. The survival was not influenced whether N2 lymph node metastases were observed at one or more level. In the multivariate survival calculations sex (p = 0.024), histological type (p = 0.006), pT classification (p = 0.002) and the pN classification (p < 0.0001) proved to be independent prognostic factor. The pM status did not affect the survival. Postoperative complications were observed in 242 of patients (28.2%). The most frequent complication were expansions problems (9.7%), bronchial stump insufficiency (3.3%), retention of bronchial secretion (2.1%) and cardiac complications (7.1%). 26 patients died, the 30-day mortality rate was 2.8%. 32 reoperations were necessary (3.7%). CONCLUSIONS: Our results indicate that non small-cell lung cancer can be operated on with good survival possibilities and limited risk, in selected cases even with more advanced tumour stages. PMID- 15803877 TI - [Change of attitude in the surgical treatment of lung metastases]. AB - There has been no major change in surgical technique of lung metastasectomy till the last decades. In the latest years the variety of diagnostic tools and methods grew significantly. There are more chemotherapy possibilities and more palliative surgical techniques (VATS, ITP, RFTA) to treat certain types of lung metastases. Based on data of the International Registry of Lung Metastases the most important prognostic factor for metastasectomy is how radical the procedure was (clear resection margin). The more aggressive approach (even pneumonectomy for R0 resection) is becoming more popular. PMID- 15803876 TI - [Our experience with patients operated after neoadjuvant therapy]. AB - The authors performed thoracotomies on 47 patients because of NSCLC between 1 January 2000 and 31 December 2003. All patients had neoadjuvant therapy which was indicated by IIIA stage NSCLC with N2 nodal status. After the neoadjuvant therapy almost all tumors (92.7%) became resectable. The combinations of therapy types, the operations type and the surgical complications are analysed. Long term outcome proves, that multimodal therapy of NSCLC (in IIIA stage) is an effective treatment method. PMID- 15803878 TI - [The role of VATS in the diagnosis and treatment of mediastinal masses]. AB - We performed 158 operations for mediastinal lesions in general anaesthesia from 1st December 1995 to 31st May 2004 using video assistance. Among them 105 procedures were diagnostic and 53 therapeutic. There was no mortality and no serious complications. We believe that VATS technique provides a big help in the diagnosis of mediastinal diseases and in the treatment of the benign lesions. It is a responsible safe method and causes less stress for the patients. PMID- 15803879 TI - [Emergency video-assisted thoracoscopic surgery for intrathoracic foreign bodies]. AB - The aim of this retrospective study is to highlight the advantages of video assisted thoracic surgery (VATS) in the assessment of thoracic trauma. In the past 4 years 3 patients had intrapleural or intrapulmonary Kirschner needles which penetrated from the site of a previous humeroscapular osteosynthesis and one patient with a gunshot projectile and haemothorax in the left thorax. They were treated by VATS removal of intrathoracic foreign bodies. No complications developed postoperatively, the chest wall pain was minimal, the duration of hospital stay (3-5 days) and recovery was short. The widespread use of minimal invasive VATS procedure is an alternative approach that provides safe and less invasive operation for acute chest trauma patients. We believe that its extended use for both diagnostic and therapeutic purposes even in some special circumstances is justified today. PMID- 15803880 TI - [The role of video-endoscopy in the treatment of thoracic empyema]. AB - The authors present their results in treating 103 cases of empyema thoracis, phase II, the fibropurulent stage, by video-endoscopy (Video-Assisted Thoracic Surgery, VATS). They highlight in which phase this method can be successful, depending on the status of the empyema, on the basis of their experiences over a ten-year period. They find its use superfluous in phase I, the exudative phase, because a suitable antibiotic and suction drainage treatment is sufficient. In phase III of empyema thoracis the use of VATS proved to be unsuccessful. In phase II they prefer video-thoracoscopy to fibrinolytic treatment. PMID- 15803881 TI - [Lung biopsy using mediastinoscope in the time of VATS]. AB - Patients with diffuse lung disease need lung biopsy for accurate diagnosis and treatment. Both traditional open lung biopsy through a thoracotomy and video assisted thoracoscopic lung biopsy are effective methods for obtaining parenchymal samples. The authors present their surgical method and experience. Thirty patients were operated on for lung biopsy using mediastinoscope between 1999-2003. Lung parenchymal samples were eligible for histological examination. No serious postoperative complications developed. The method is simple, safe and low-cost. PMID- 15803882 TI - [Changes in surgical therapy of pulmonary hydatid cysts]. AB - The lung is the second most common site of hydatid cysts after the liver. The authors analyse retrospectively the results of patients treated with pulmonary hydatid cysts in the past 18 years, considering video-thoracoscopy. Twenty eight patients were treated during this period in 31 cases. Hydatid disease affected only the lung in case of 22 patients, while in 6 cases it was present in the liver and lung simultaneously. Pulmonary hydatid disease affected one side in 24 and both sides in 4 cases. For surgical treatment pericystectomy in one, atypical segment resection in 18, anatomical segmentectomy in three, lobectomy in 7 and video-thoracoscopy in 3 cases were performed without surgical complications. The mean hospital stay was 10.5 days in case of thoracotomies and 8.5 days in case of video-thoracoscopy. There was one recurrence in conventional surgery and reoperation was necessary. After video-thoracoscopy no recurrence was detected. Mean follow-up was 120 months, after video-thoracoscopy it was 20 months. Three patients have uncertain chest pain after thoracotomy, but none has any complaints after video-thoracoscopy. Fifteen patients took mebendazole permanently after the final histological result. According to the authors' practice the indication of lung resections for pulmonary hydatid cysts is limited, in selective cases video thoracoscopic cystectomy can be a successful treatment of choice. PMID- 15803883 TI - [Prevention and treatment of bronchopleural fistula using pedicled muscle flaps]. AB - Authors used pedicled muscle flaps for prevention and treatment of bronchopleural fistulas in 55 patients. Thirty five flaps were used for prevention and 21 for treatment of bronchial stump leak. After using 19 pectoral, 18 latissimus, 10 diaphragm, 3 intercostals, 2 serratus and 4 double muscle flaps, only two flap necrosis occurred due to insufficient flap perfusion. Latissimus and diaphragm flaps were used mainly for the purpose of prevention and the pectoral flaps for the treatment of bronchopleural fistula. The authors reveal the advantages, disadvantages and indications of using different muscle flaps for different purposes. Using this technique is an essential practice in everyday thoracic surgery. PMID- 15803884 TI - [Role of surgical lung volume reduction in the treatment of patients with severe emphysema]. AB - OBJECTIVE: Based on literature data and own experience (unique in Hungary) we like to present the indications, contraindications, risks as well as medium and long term follow ups of the lung volume reduction surgery (LVRS) applicable in cases of therapeutic resistant end-stage emphysema (COPD). MATERIAL: In Szombathely between 1997 and 2003, 69 LVRS--by multiple wedge resections of most functionless parts of the lungs--were made on 56 patients (44 males and 12 females with a mean age of 52.5 years) via median sternotomy for both sides (12 pts), axillary thoracotomy on one (30 pts) or both sides in two sessions (13 pts) and VATS (one side, 1 pt). Selection criteria based on radiographic, spirometric and cardiac function tests. Indications were: diffuse homogenous emphysema: 17 pts, heterogenous emphysema 39 pts. Combined surgery for severe COPD and pulmonary malignancy was applied too (7/56 pts - 12.5%). RESULTS: Thirty-day operative mortality was 4.3% (3/69) similar to international results. We noticed high incidence of early complications: air leak (37.6%), serious arrhythmia (13%), respiratory insufficiency (8.7%), longer drainage time (5.7 days). Reoperations were 2/69 interventions (2.9%) because of air leak and bleeding. Late recurrent pneumothorax was observed in 1 case (1.3%). The mean follow up time was: 31.8 months. FEV1, RV, paO2, paCO2 and the quality of life improve as seen in follow-up, and patients regain apart of the former activity. There were no differences in the outcome in patients undergoing unilateral or bilateral LVRS. On the basis of literature and own data the long-term significant positive improvements last about 12-24 months. Thereafter one can observe a slow decrease of the cardiopulmonary functions, however, the quality of life remains adequate for longer. SUMMARY: The LVRS has a certain risk, but perioperative mortality is acceptable. LVRS is a realistic alternative of lung transplantation in case of severe COPD first of all in patients presenting with heterogenous emphysema (particularly upper lobe) and low exercise capacity. The expense of LVRS is maximum 10% of lung transplantation. Patients having undergone an LVRS and patients unacceptable for LVRS can be suitable for lung transplantation. PMID- 15803885 TI - [Evidence based therapy with insulin in diabetic patients]. AB - A fast development in therapy with insulin was observed after its discovery. Besides the widely used human regular insulin preparations, nowadays ultrashort and long-acting insulin analogues are also available for the patients. At present, the results of large clinical trials enable an evidence based diabetes care. It is well documented, that near-normoglycemia should be achieved by intensive conservative insulin treatment or pump therapy in type 1 diabetic patients. The beneficial effects of the good metabolic control could also be observed years later concerning late specific complications of diabetes. Similarly, as good as possible metabolic control should be aimed with antidiabetic treatment including insulin, if necessary, in type 2 diabetic patients. It is documented that the risk of cardiovascular complications is not increased in type 2 diabetic patients treated with insulin. Hypoglycemia and weight gain are the most important side effects of the insulin treatment. Recently, evidence based recommendations for treatment with ultrashort (insulin lispro, insulin aspart) and long-acting insulin analogues (glargine) can also be determined. PMID- 15803886 TI - [Congenital heart malformations is twin pregnancies]. AB - INTRODUCTION: In twin pregnancies the risk for congenital heart malformation is higher than in singular pregnancies. Because of the development of prenatal diagnostic methods, it is possible to recognise congenital malformations. In point of view of congenital cardiac malformations the twin pregnancies belong to high risk group. AIM: To found connection between congenital heart malformations and twin pregnancies. MATERIAL AND METHOD: Retrospective analysis of data of Fetal Cardiology database between 1. january 1966 and 30. november 2003. RESULTS: In singular pregnancies 455, and in twin pregnancies 31 severe congenital heart malformations were diagnosed prenatally. It means, that congenital heart malformations were diagnosed prenatally in 2% of singular pregnancies and in 4,6% of twin pregnancies. In monozygotic twin pregnancies in 36% of heart malformations were pulmonary stenosis and in 45% endocardial fibroelastosis. In dizygotic twin pregnancies Ebstein malformation was more common than it statistically expected. In dichorial and dizygotic twin pregnancies the cardiac malformations were similar to in singular pregnancies. CONCLUSIONS: In twin pregnancies the rate of congenital heart malformations is higher than in singular pregnancies, that's why the twin pregnancy is indication for fetal echocardiography. In monochorial twin pregnancies different types of congenital heart malformations are expected than in singular or dichorial twin pregnancies, the chorionicity seems more important than the zygosity. PMID- 15803887 TI - [22q11 deletion in conotruncal anomalies]. AB - 22q11 deletions in conotruncal anomalies. Two children are reported in whom conotruncal anomalies (truncus arteriosus communis, tetralogy of Fallot) were associated with chromosome 22q11 deletion. In both cases, which represent the first published cases in Hungary according to the knowledge of the author, deletions were suspected on the basis of phenotype termed Di George syndrome. Nowadays the role of molecular genetics is growing in the etiology of congenital heart defects and the chromosome 22q11 deletions constitute one of the most frequent genetic mutations associated with congenital heart defects. The author emphasizes that clinicians must know about this disease, and in case of suspicion they should request for molecular genetic investigation. 22q11 deletions have an important roles in the prognosis of congenital heart disease, in counseling and in the prenatal diagnosis as well. PMID- 15803889 TI - [Treatment of renal tumors]. PMID- 15803888 TI - [Pneumatosis cystoides intestinalis, as a rare complications of coeliac disease]. AB - Intestinal pneumatosis cystoides is rarely diagnosed clinical entity characterized by multiple gas containing cysts inside the wall of the gastrointestinal tract. Primary (15%) and secondary (85%) forms are known. In the more frequent secondary forms small intestine and the right side of the colon are mainly affected, and the underlying pathology can be gastrointestinal, pulmonary or immunological. The 64 year old male patient reported by the authors showed clinical signs of severe malabsorption (his body weight: 47,5 kg, height: 178 cm, BMI: 15). The intestinal pneumatosis was diagnosed by exploration performed because of suspected perforation (pneumoperitoneum). Due to severe malabsorption the patient was admitted to Gastroenterological Department. Duodenoscopy and small bowel biopsy was performed. Small intestinal histology and presence of anti gliadin and anti-endomysium antibody confirmed the suspected diagnosis of coeliakia. Parenteral nutrition, special gliadin free diet, hyperbaric oxygen inhalation, steroid, and metronidazole treatment resulted in a gradual improvement in the nutritional and general condition of the patient. After one year the patient is without complaints, he gained 24,5 kg body weight BMI: 22,5). The author's case points to a rare serious complication of non-treated coeliakia. Recognition of intestinal pneumatosis in time could have prevented surgical intervention in the high risk patient. PMID- 15803890 TI - [Ferenc Foldes (1910--1997)]. PMID- 15803891 TI - [Memory of Pal Peterffy M.D]. PMID- 15803892 TI - [Early results of coronary artery bypass reoperations in own experience]. AB - Coronary Artery Bypass Grafting (CABG) is widely accepted as an effective and durable method of treatment of Coronary Artery Disease. In some patients however, recurrent angina and the necessity of reoperation (RE-CABG) occurs. AIM: The aim of the study is the estimation of the results of RE-CABG in own material. MATERIAL AND METHODS: In years 1995-2002 CABG operations were performed in 3452 patients (CABG group) and RE-CABG in 37 pts (1.07%) (RE-CABG group). Patients were considered for operation on condition of the presence of symptomatic CAD confirmed by coronarography in standard projections. Operations were performed using Cardiopulmonary Bypass. RESULTS: The CABG group consisted of 3452 pts in mean age 57.9 years, including 2693 men (78%) and 759 (22%) women. The RE-CABG group was composed of 37 pts in mean age 60.7 years, comprising 22 (59.5%) men and 15 (40.5%) women. The percentage of women in CABG group was significantly higher than in CABG group. The mean age in RE-CABG group was insignificantly higher than in CABG group. In the CABG group 3.1 anastomosis/pt were performed, including anastomosis IMA to LAD in 2779 pts (80.5%). From this group only in 2 pts (0.07%) coronary reoperations were performed. In the remaining 673 pts from CABG group during the first operation, only saphenous vein graft were performed and in 35 (5.2%) coronary reoperation was necessary. This difference is statistically significant. In-hospital mortality in CABG group was 5%, the IABP was used in 7.8% of pts. Mortality in the RE-CABG group reached 8.1% (3/37), the IABP was used in 16.2% (6/37). CONCLUSIONS: 1. The use of arterial grafts during the first coronary operation decreases the reoperation rate. 2. RE-CABG are performed in the older patient rather than CABG. 3. RE-CABG are performed more frequently in women than in men as compared to CABG. 4. RE-CABG are burdened with twice as high mortality rate and the necessity of use of IABP than CABG. PMID- 15803893 TI - [Is Q-wave on admission associated with microvascular injury in patients with acute myocardial infarction treated with primary angioplasty?]. AB - INTRODUCTION: Impaired microvascular flow, despite patent epicardial artery (no reflow phenomenon), leads to greater left ventricular dysfunction after myocardial infarction (MI). Predictive factors associated with no-reflow remain largely unexplored. Q-wave on admission (Q(A)) is a sign of extensive ischemia probably predisposing to no-reflow. The aim of the study was to explore possible relation between Q(A) and electrocardiographic signs of no-reflow in patients with first MI. MATERIAL AND METHODS: The study group was composed of 108 patients (81 men; mean age 60+/-11 years), with first ST-segment elevation MI, treated successfully with primary angioplasty (p-PTCA). ECG tracings were obtained before and 30 minutes after p-PTCA. The sum of ST-segment elevations (sum(ST(el))) in 3 contiguous leads with the highest ST(el) was calculated. Lack of 50% reduction of the sum(ST(el)) 30 minutes after angioplasty was defined as ECG sign of no reflow. Presence of Q(A) was estimated in leads with ST(el). RESULTS: Q(A) was found in 42 (39%) patients. Q(A) was more often observed in patients with ECG signs of no-reflow (38% vs. 18%; p<0.05). Group with Q(A) showed larger damage of left ventricle estimated with ECG QRS score (7.7+/-4.4 vs. 6.1+/-3.4; p<0.05) as well as worse ejection fraction (42% vs. 46%; p=0.05). CONCLUSIONS: Patients with Q(A) have more often ECG signs of no-reflow than other patients with MI. Previously described worse function of left ventricle in this group, may be partially caused by more frequent no-reflow occurring in those patients. This fact suggests that adjunctive therapy preventing no-reflow could be beneficial in this group of patients. PMID- 15803894 TI - The effect of chronic alcohol consumption on systolic and diastolic left ventricular function. AB - The study population consisted of 248 patients with at least five-year (mean 8.7+/-4.2 yrs.) history of drinking, mean age (48+/-4.2 years), including 132 subjects with arterial hypertension (mild or moderate according to WHO/ISH guidelines) and 116 normotensives. A group of 48 patients with essential arterial hypertension not consuming alcohol served as controls. Groups of alcoholics with hypertension, alcoholics with normal blood pressure and the controls were compared with respect to differences in standard BP measurements and 24hr blood pressure monitoring, left ventricular mass index, systolic and diastolic left ventricular function by echocardiography. In alcohol-dependent subjects the left ventricular mass index progressed with the increasing declared consumption of ethanol and duration of abuse. The incidence of LVH in drinking hypertensives (28%) did not differ from that in non drinking hypertensives (26%), p>0.05. In hypertensives irrespective of ethanol consumption delayed relaxation was a marker of left ventricular diastolic dysfunction. The present study did not confirm the hypothesis that left ventricular hypertrophy was responsible for diastolic impairment in alcohol abusers. Chronic ethanol consumption significantly deteriorated left ventricular diastolic function irrespective of its effect on blood pressure and left ventricular mass. In patients with the highest level of ethanol consumption and the longest duration of abuse ejection fraction was significantly lower than in patients with the lowest level of alcohol consumption and the shortest duration of abuse (EF 61.8% vs. EF 67.4%, p<0.05). Thus, the deterioration of systolic function is significantly related with the level of alcohol consumption and duration of abuse. PMID- 15803895 TI - [Factors affecting the cooperation of hypertensive patients in the process of treatment]. AB - INTRODUCTION: Arterial hypertension is one of the leading causes of cardiovascular morbidity and mortality. Despite rigorous knowledge of pathophysiology of this entity and wide accessibility to highly effective drugs the rate of hypertension control is still low. Currently the lack of appropriate patient cooperation including irregular drug taking and unhealthy lifestyle are regarded to contribute most to the low efficacy of hypertension treatment. Many studies demonstrated the role of educational programmes in better hypertension control. It is now necessary to indicate those groups that need such efforts particularly. AIM: The aim of this study was to assess the factors affecting regular drug taking among hypertensive patients. METHODS: 222 ambulatory patients (x=56 years, 123 males and 99 females) of the I Department of Cardiology of the Jagiellonian University Medical College in Krakow were analyzed in this study. We used a detailed, uniform own questionnaire to get the following information from the patients: regularity of drug taking, age, level of education, place of living, occupational category, incomes, coexisting diseases, costs of drugs and self-control of blood pressure. Afterwards we compared two groups: patients that use drugs systematically and those who do not. A computer software Statistica PI was used for statistical analyses. RESULTS: Most of patients (129; 58%) declared regular drug taking. We observed statistically significant differences between two analyzed groups for the following parameters: level of education, place of living, occupational category, incomes, knowledge of the definition of hypertension and regularity of blood pressure self-control. Patients living in cities, currently working, with higher level of education, higher incomes, well defining hypertension and measuring blood pressure regularly were more systematic in drug taking. The logistic regression analysis showed that only low incomes were independently associated with irregular drug using. PMID- 15803896 TI - [The role of serum troponin I and high sensitivity C-reactive protein concentrations in early diagnosis of acute coronary syndrome in patients admitted to Emergency Unit]. AB - The aim of this study was to show clinical benefit of simultaneous determinations of troponin I (cTnI) and high sensitivity C-reactive protein (hsCRP) at early diagnosis in the group of patients with stenocardia admitted to the Emergency Unit of 5-th Military Hospital in Krakow. The study group consisted of 38 (51.3%) patients with acute myocardial infarction (AMI) and 36 (48.6%) with unstable angina pectoris. ROC function analysis for cTnI (cutoff value 0.69 ng/mL) and hsCRP (3.33 mg/L) indicated significant amelioration of diagnostic sensitivity into 84.9% at early diagnostics of AMI in group of patients with acute coronary syndrome (ACS). Moreover, studies of mutual relationship between elevated levels of hsCRP and cTnI concentrations have shown a remarkably positive correlation (r = 0.90; p < or = 0.01) in the group of patients with AMI. PMID- 15803897 TI - [Periodontal and dental state of patients with coronary heart disease]. AB - INTRODUCTION: Nowadays periodontitis--etiologically linked with bacterial flora, mainly anaerobic--is very often pointed out as one of potential risk factors for coronary heart disease (CHD) and myocardial infarction. Only few Polish publications on this subject are available. The aim of this study was to evaluate the frequency and degree of advancement of periodontitis among patients with CHD and after myocardial infarction, hospitalized in I Clinic of Cardiology, Medical University in Gdansk, Poland. Simultaneously an attempt to answer the question, if periodontitis can be a potential risk factor for CHD and myocardial infarction in Poland's population. MATERIAL AND METHODS: 104 patients, aged 43-80 (average 60 years) were examined--57 persons after myocardial infarction (group I) and 47 persons with CHD without acute events (group II). Anamnesis and clinical examination using WHO recommended parameters (API, SBI, PD, CAL, CPITN) were performed. RESULTS: In 82.4% patients of group I and 72.2% of group II periodontitis was diagnosed. This percentage was significantly higher in comparison with general population. Advancement of destructive lesions suggests long duration of disease and it's possible negative influence on general health status. The results indicate possible link between periodontal infection's foci and CHD, and also show, that education, prevalence and treatment program for this population should be created. PMID- 15803898 TI - [Costimulatory and activation molecules of lymphocytes T in acute lymphoblastic leukemia in children]. AB - In the last years one rises importance of costimulatory molecules in immune response in leukemias. Aim of the study was to assess lymphocytes T function in children with acute lymphoblastic leukemia during remission induction on the grounds of chosen costimulatory and activatory molecules expression. To assess percentages of lymphocytes subpopulations we used tricolor flow cytometry. RESULTS: 1. In the moment of diagnosis and remission induction we noted higher percentages of lymphocytes T with adhesion molecule ICAM-1; 2. During remission induction we observed lower percentage values of lymphocytes T with CD38 coexpression; 3. In the end of remission induction rised the percentage values of lymphocytes T helper with IL-2 receptor expression; 4. In the group of patients with fever/infection we observed higher percentage of activated lymphocytes T (CD3-HLA-DR) comparing to non-infected patients. Summarizing, we suggest lymphocytes T activation during appearance and remission induction of acute lymphoblastic leukemia in children. This confirms participation of cellular immunity in leukemic process. PMID- 15803899 TI - [Th1/Th2 balance in acute lymphoblastic leukemia in children]. AB - Analysis of T lymphocytes cytokine profiles allows to differ subpopulations: Th1, Th2, Th3, Tr1. Aim of the study was to assess Th1/Th2 balance in acute lymphoblastic leukemia in children at diagnosis and during/after remission induction, especially during infections. Percentages of lymphocytes T producing IFN-gamma and IL-4 were assessed by flow cytometry. We noted the rise of lymphocytes T helper producing IFN-gamma (Th1) and percentage of lymphocytes T producing IL-4 at the beginning and during remission induction was higher than in control group. During fever/infection we observed the rise of lymphocytes Th1, and no change in Th2 percentage. Summarizing we suggest Th1/Th2 imbalance and Th2 predominance in acute lymphoblastic leukemia in children. PMID- 15803900 TI - [Usefulness of the "P.N." frame in biopsies and stereotactic operations on the brain]. AB - BACKGROUND: Stereotactic biopsy represents the gold standard for tissue diagnosis surgically inaccessible gliomas, tumors and cystic lesions. Tissue samples can be obtained by minimally invasive technique. Moreover, the combination of the neuro endoscope with stereotactic method is possible. MATERIAL AND METHOD: 23 patients underwent 43 stereotactic biopsy procedures for diagnosis diffuse, deep-seated and eloquent site tumors at Neurosurgery Department Jagiellonian University in Krakow since February 2003. Stereotactic frame "P.N." type (TatraMed S.r.o.) was used. In one case the combined method was used for third ventriculostomy. The specimen was obtained from the target point of CT scan by the aspiration method under local anesthesia in 17 patients and under general anesthesia in 6 patients. The lesion was located in 9 cases of cerebral hemisphere and basal ganglia, in 8 cases of spreading site, in 5 cases of eloquent area and in 1 case of corpus callosum and third-ventricle. RESULTS: Material from the stereotactic biopsy has been examined neuropathologically. Astrocytoma II(o) WHO were diagnosed in 6 cases, Astrocytoma III(o) WHO in 11 cases, Astrocytoma IV(o) WHO in 3 cases, Metastatic adenocarcinoma in 1 case and in 2 cases the neoplasm was not found. The diagnosis was histologically confirmed in 91.3%. Emergency craniotomy was performed in one patient. In 3 cases the increased intracranial pressure produced by tumor cysts was reduced by the use of biopsy. CONCLUSIONS: Our data confirm that the "P.N." frame is useful in stereotactic biopsies and operations even with the use of neuroendoscope. The procedure is safe, easy to follow as a tool for patients with various tumors. PMID- 15803901 TI - [Transsphenoidal surgery of secretory and invasive pituitary adenomas (somatotropinoma or corticotropinoma)--own experiences]. AB - Invasive pituitary adenomas with extension to the cavernous sinus are rare and comprise 6-10% of all pituitary tumors. Their effective surgical treatment is very difficult. Cavernous sinus invasion increases the morbidity and mortality associated with surgical procedures. Prolonged medical treatment and radiotherapy are an alternative to the surgery. PURPOSE: Efficiency of the extended transnasal transsphenoidal approach in the treatment of the invasive pituitary adenomas with extension to the cavernous sinus. MATERIAL AND METHOD: During the past two years (from January 2000 to December 2001) authors carried out 24 surgical operation in the medial compartment of the cavernous sinus (extended transnasal transsphenoidal approach). Our experience is based on the analysis of 24 patients (19 patients with GH-secreting tumors and 5 patients with ACTH-secreting adenomas -2 with Cushing disease and 3 with Nelson's syndrome) with invasive macroadenomas expanding to the cavernous sinus. Mean age was 46.21 years (range 28-68, SD +/ 9.54 years) in our group. Mean serum GH level was 43.95 microg/l (range 9-170 microg/l, SD +/-39.68 microg/l) and mean serum ACTH level in the group of Nelson's syndrome was 1180 pg/ml (range 790-1780 pg/ml, SD +/-527.35 pg/ml). Parasellar extension of the tumor was measured according to Knosp's scale. There were 16 patients in stage III and 9 patients in stage IV. RESULTS: Total surgical removal of invasive GH-secreting adenoma (according cure criteria: basal serum GH level below 2.5 microg/l, OGTT<1 microg/l, normal sex and age-related IGF-I level) we achieved in 6 patients. Basal serum GH level was below 5 microg/l (postoperative mean serum IGF-I level was significantly lower but abnormal) in the next five patients. Remission (normalization of 24-hour urine-free cortisol (UFC) and its metabolites) was achieved in one patient with Cushing disease. Out of 3 patients with Nelson's syndrome two were cured. Their postoperative serum ACTH level was below 80 pg/ml. There were no mortality and no deterioration of cranial and optic nerves. CSF leakage occurred in one patient and was treated surgically. Deterioration of pituitary function and diabetes insipidus occurred in one patient. CONCLUSION: Extended transnasal transsphenoidal approach is the safe surgical procedure in the treatment of invasive GH or ACTH-secreting pituitary adenomas. The overall morbidity rate of this surgical intervention is rare. We suppose the extended transnasal transsphenoidal approach is an alternative for prolonged medical treatment and radiotherapy. PMID- 15803902 TI - [Selectins in the pathogenesis and diagnosis of early atherosclerotic changes in children and adolescents with risk factors (obesity, hypertension and diabetes)]. AB - Selectins are the group of adhesion molecules, which main role is the tethering of leukocytes to the endothelium. They induce relatively weak and transient adhesion allowing the cells to roll along the vascular wall. That mean, that selectins play part in the earliest stages of the atherosclerotic process. The aim of the study was to evaluate levels of E-selectin, L-selectin and P-selectin in children and adolescents with traditional atherosclerosis risk factors (obesity, hypertension and diabetes), correlation with lipid parameters, blood pressure values and BMI as well as the attempt to answer the question whether selectins, and which of them can be useful in predicting cardiovascular risk in these young patients. MATERIAL AND METHODS: We studied 66 children aged 14.8+/ 2.6 years. They were divided into 4 subgroups according to the diagnosis: children with obesity--OP (n=12), children with obesity with hypertension--OP and NT (n=14), children with hypertension--NT (n=12), children with diabetes--C (n=28). Control group consisted of 15 healthy, slim children and adolescents, aged 15.4+/-2.2 yrs. Levels of E-selectin, P-selectin and L-selectin were evaluated by immunoenzymatic methods with use of R&D Systems ELISA kits. RESULTS: In the study group we found significantly higher level of E-selectin--86+/-28 ng/mL compared to control group: 64+/-20 ng/mL (p<0.05). Similarly in all subgroups levels of E-selectin were significantly higher: in obese children E selectin level was 95+/-31 ng/mL (p<0.05), in obese and hypertensive group--88+/ 12 ng/mL (p<0.05), in hypertensive group--79+/-19 (p<0.05), and in diabetic children--83+/-25 ng/mL (p<0.05). L-selectin level in the study group was 1552+/ 290 ng/mL and did not differ from control group--1652+/-281 ng/mL (ns). P selectin level in the study group was 652+/-419 ng/mL and was similar to the level of P-selectin in the control group--669+/-295 ng/mL (ns). Correlation analysis showed significant relationship between E-selectin and BMI (r=0.23, p<0.05) and diastolic blood pressure (r=0.24, p<0.05). CONCLUSIONS: 1. Young patients with atherosclerosis risk factors have elevated level of E-selectin. 2. E-selectin level correlates with BMI and diastolic blood pressure. 3. Elevated level of E-selectin may confirm endothelial dysfunction in young patients with atherosclerosis risk factors and can serve as a marker of early atherosclerosis phases. PMID- 15803903 TI - [The quality of life of diabetic patients--literature review]. AB - The knowledge concerning the influence of psychosocial factors on the functioning of patients with diabetes seems still insufficient. In the article, the scales measuring global quality of life and the effect of the illness on the quality of life, as well as the results of studies on quality of life of diabetic patients are presented. Improvement of quality of life may have both medical and economic consequences. PMID- 15803904 TI - [Symptoms and treatment of depression in patients with diabetes]. AB - The prevalence of depression in patients with diabetes may be as high as 30%. Results of studies point to insufficient diagnosis and treatment of depression in diabetic patients. Guidelines for diagnosis and treatment of depression occurring in diabetic patients are presented. PMID- 15803905 TI - [Uric acid and diabetes mellitus]. PMID- 15803906 TI - [Glucagon-like peptides--synthesis, biological actions and some clinical implications]. AB - Glucagon and the glucagon-like peptides (GLPs) are derived from single proglucagon gene and exhibit an increasing number of biologically important actions. As a counter-regulatory hormone for insulin, glucagon plays a critical role in maintaining glucose homeostasis in vivo in both animals and humans. To increase blood glucose, glucagon promotes hepatic glucose output by increasing glycogenolysis and gluconeogenesis and by decreasing glycogenesis and glycolysis in a concerted fashion via multiple mechanisms. The glucagon-like peptides GLP-1 and GLP-2 are produced in enteroendocrine L cells of the small and large intestine and secreted in a nutrient-dependent manner. GLP-1 regulates nutrient assimilation via inhibition of gastric emptying and food intake. GLP-1 controls blood glucose following nutrient absorption via stimulation of glucose-dependent insulin secretion, insulin biosynthesis, islet proliferation, and neogenesis and inhibition of glucagon secretion. Glucagon-like peptide-1 (GLP-1 is an insulinotropic hormone, GLP-1 also inhibits glucagon secretion. GLP-1 lowers blood glucose in normal subjects and in patients with type 2 diabetes. The major biological action of GLP-2 appears to be the stimulation of small-bowel hyperplasia, manifested by an increases in both villous height and small-bowel weight. A pilot study of GLP-2 administration in human subjects with short bowel syndrome demonstrated significant improvements in energy absorption, bone density, increased body weight, which correlated with increased crypt plus villus height on intestinal biopsy sections. The biological actions of two of these glucagon-related peptides, suggest that they may have therapeutic relevance for the treatment of human diseases such as diabetes, selective intestinal disorders and cardiac diseases. PMID- 15803907 TI - [The role of chemokines in the pathogenesis of restenosis]. AB - In the article the role of immunologic cells and hemostatic factors in the formation of atherosclerosis was described. Attention was paid to restenosis in patients undergoing percutaneous transluminal coronary angioplasty (PTCA), a phenomenon that occurs in 20-50% of these patients. Among many factors that contribute to restenosis more attention is paid lately to the role of chemokines. The physiology role of chemokines was described and the contribution of monocyte chemoattractant protein-1 to restenosis after PTCA. PMID- 15803908 TI - [New beta-blockers in the treatment of chronic ischaemic heart disease]. AB - Beta-blockers are routinely used in all forms of ischaemic heart disease except variant angina. Recently, III generation beta-blockers were introduced into practice. New beta-blockers often have ancillary properties like vaso-dilating or antioxidant properties. Also slow-release formulas of old beta-blockers are now available. Several studies show that these new agents are more efficient when compared with old beta-blockers. Moreover, adverse effects of new beta-blockers are often less severe than in the case of older agents. Especially the influence on bronchi, peripheral arteries and lipid and glucose metabolism is less pronounced. The frequency of beta-blockers use is too low in Poland as well as in other European countries, despite overwhelming evidence that these agents improve prognosis. The results of Cracovian Program for Secondary Prevention of Ischaemic Heart Disease showed that the proportions of patients after myocardial infarction or myocardial revascularization prescribed beta-blockers are too low. One of the reasons is probably the doctors' concern about side effects of beta-blockers. Widespread use of new beta-blockers in everyday clinical practice may increase beta-blocker use in general and in this way improve prognosis of ischaemic heart disease patients in Europe. PMID- 15803909 TI - [Diastolic dysfunction as a risk factor in patients with heart failure]. AB - In the recent decade diastolic dysfunction has been extensively studied. It has been found that the clinical syndrome of heart failure may result from two entirely different disorders--systolic and diastolic impairment or both. Available evidence indicates that the left ventricular filling pattern is altered with progressing diastolic dysfunction. Left ventricular diastolic dysfunction is a sensitive marker of myocardial damage and it can be used to monitor a spectrum of changes ranging from impaired relaxation through pseudo-normalization to restrictive filling pattern. The restrictive filling pattern, irrespective of its etiology, has a negative effect on prognosis, especially when it coincides with reduced left ventricular ejection fraction. Patients with systolic heart failure and restrictive filling pattern show a lower exercise tolerance than their counterparts with non-restrictive filling. This indicates a significant role of diastolic dysfunction in the development of clinical symptoms in patients with left ventricular systolic impairment. Progress in the understanding of diastolic dysfunction may allow for better evaluation of prognosis and therapeutic efficacy and for monitoring of patients with systolic dysfunction. PMID- 15803910 TI - [Arterial hypertension in pregnant women]. AB - In the paper we described the criteria of diagnostics and divisions of hypertension in pregnant women. We noticed the possible etiology, symptoms and complications of the disease both for fetus and mother. We described the treatment of hypertension during pregnancy as well. PMID- 15803911 TI - [Sarcoidosis--clinical features, diagnosis and treatment]. AB - Sarcoidosis is a multisystem disorder of unknown aetiology. Pathologically it is characterized by the presence of non-caseating epithelioid-cell granulomas in the lungs, intrathoracic lymph nodes and other affected organs. The clinical course of sarcoidosis is widely variable, ranging from asymptomatic but abnormal findings in chest radiography to progressive multiorgan disease, which leads to fibrosis with organ failure and functional impairment. The aim of this article is to familiarize physicians with clinical features and treatment of this disease. PMID- 15803912 TI - [Diagnostic value of procalcitonin (PCT) determination]. AB - A large number of publications, primarily clinical studies, demonstrate the increasing use of procalcitonin (PCT) in modern clinical practice. PCT is a propeptide of calcitonin induced by a variety of stimuli including bacterial endotoxins, proinflammatory cytokines and triggering events such as trauma or cardiogenic shock. PCT is not or only slightly induced by viral infections, autoimmune disorders, neoplastic diseases and organs transplantation. The aim of this study was to evaluate the usefulness of PCT as a diagnostic and prognostic tool in patients for early differentiation between bacterial and nonbacterial infection origin. PMID- 15803913 TI - [Acute pancreatitis in pregnant women--case reports]. AB - Two patients with acute pancreatitis were presented. In the first case the disease occurred in the 32 week of pregnancy and in the perinatal period in second case. Some changes in lipid metabolism and composition of the bile in pregnant women and their role in pathogenesis of acute pancreatitis are discussed. PMID- 15803914 TI - [Adaptation of different neuroprotection devices during carotid stenting procedures]. AB - Carotid artery stenting (CAS) is, apart from surgical endarterectomy, a common method of treatment of atherosclerosis in carotid arteries. Percutaneous angioplasty of carotid artery narrowings has been developing in recent years due to introduction of neuroprotection systems, which significantly reduced number of complications. Nowadays three neuroprotection systems are in common use: filters, temporary occlusion and aspiration systems and flow reversal systems. We present three cases of successful CAS with application of three different neuroprotection systems. We discuss indications, contraindications and limitations of every device. PMID- 15803915 TI - [Auricular perichondritis in adults. An evaluation-study of 12 hospitalized patients]. AB - Perichondritis of the auricle in adults can be secondary to traumatic agents, external otitis or surgery being Pseudomona aeruginosa the bacteria more often isolated. We have performed a retrospective study of 12 hospitalized patients with that diagnosis and these following variables have been evaluated: Age, sex, personal antecedents, clinical symptoms, affected auricle, complementary explorations, treatment, evolution and stay. PMID- 15803916 TI - [Prognosis value of the immunohystochemical expresion of the p21WAF1 in the larynx cancer]. AB - In this paper we carried out an immunohistochemical study of protein p21WAF1 (EA10) expression in a series of 195 patients with laryngeal carcinoma that were diagnosticated, treated and followed at the Department of Otolaryngology at "Virgen de la Salud" Hospital (Toledo, Spain). In the cases with lymph node metastasis we also studied p21WAF1 expression at this level. Furthermore we have analysed the value of protein p21WAF1 expression as a prognostic factor (tumor recurrence, deads due to cancer and survival) and we evaluate the relationship between p21WAF1 expression and other clinic and pathologic parameters. PMID- 15803917 TI - [Presentation of a case of cervical actinomycosis versus Hodgkin lymphoma]. AB - Actinomycosis is an unfrequent bacterian infection, that affects 50% of the cases in the cervical facial region. The evolution and clinical, anatomopathologic radiologic characteristic, and its presentation bring us to suspect another type of more frequent pathology: the neoplastic, that make very difficult its diagnosis. We present the case of a young patient affected by a cervical tumoration initially oriented as Hodgkin lymphoma that finally was diagnosed as cervical actinomycosis and treated with penicillin. PMID- 15803918 TI - [Prognosis value of the immunohistochemical expresion of the bcl-2 in the larynx epidermoid cancer]. AB - In this paper we carried out an immunohistochemical study of bcl-2 protein expression in a series of 195 patients with laryngeal carcinoma that were diagnosticated, treated and followed at the Department of Otolaryngology at "Virgen de la Salud" Hospital (Toledo, Spain). In the cases with lymphonode metastasis we also analysed bcl-2 protein expression at this level. Furthermore we have studied the value of bcl-2 protein expression as a prognostic factor (tumor recurrence, deads due to cancer and survival) and we analysed the relationship between bcl-2 protein expression and other clinic and pathologic parameters. PMID- 15803919 TI - [Laryngeal tuberculosis masquerading as carcinoma. Report of one case and literature review]. AB - Since the introduction of antituberculous medications, the incidence of laryngeal tuberculosis (LTB) has decreased. However, it is a fact the increasing number of cases of pulmonary or extrapulmonary TB mainly caused by the Acquired Immunodeficiency Syndrome. We report the clinical case of a 72 years old male with dysphonia secondary to LTB which masqueraded a carcinoma. It's certain diagnosis was the histopathologic exam of the biopsy obtained by microsurgery. There was a residual TB in lung after a complete study of Internal Medicine Department. PMID- 15803920 TI - [Tuberculous epiglotittis, an atypical form of laryngeal tuberculosis. Presentation of a case and revision of litterature]. AB - An important arousal of tuberculosis has been observed in the last years, together with a change in it's clinical patterns, specially the extrapulmonar form, among wich is this laryngeal tuberculosis. This two facts and the marked polymorphism and mimetism proper of this condition may suggest that the Mycobacterium tuberculosis should be included in the differential diagnosis of any atypical lesion of the larynx, in order to obtain an earlier diagnosis excluding carcinoma and instaurate a chimotherapic protocol for minimizing or even avoiding any sequelae. Tuberculous epiglotittis is one of these atypical and unusual forms. The authors report a case of a 67 year-old male and provide a discussion on the diagnostic problems of this condition. PMID- 15803921 TI - [Granular cell tumor of the tongue (Abrikossoff's tumor). Report of a case and review of the literature]. AB - Granular cell tumor, myoblastoma or Abrikossoff's tumor is an uncommon benign lesion, usually of slow-growing, that shows a predilection for the oral cavity, especially the tongue. Its real etiopathogenia remains still controversial and in it's histogenesis have been implicated several cell types. We report the case of a 18 years old woman with that diagnosis, confirmed in AP, who was operated by removal of the tumoration that was located in the left half-tongue. After 28 months she has not suffered any recurrences. PMID- 15803922 TI - [Glossopharyngeal neuralgia. A propos of one clinical case]. AB - Glossopharyngeal or the 9th cranial nerve neuralgia is an uncommon pathology characterized by severe paroxysmal attacks of pain in the base of the tongue, posterior pharynx and tonsillar fossa some times associated with pain irradiated to the ear. It's Carbamazepine is the first choice of medical treatment while the microvascular decompression is considered the main surgical procedure. PMID- 15803923 TI - [Double skin paddle fibula free flap in orofacial reconstruction. Case report]. AB - Mandibular reconstruction in cases of complex muco-cutaneous defects is a challenge for head and neck surgeons. Here, we report the case of a patient who showed an oro-facial defect including bone, skin and mucosa. We decided to use a double skin paddle fibula osteocutaneous free flap for the reconstruction. Identification and preservation of the septo and musculocutaneous perforators vessels to the skin is the clue step to ensure flap vitality. In this situation mucosal and cutaneous reconstruction is possible using a folded skin paddle. We also discuss the reconstructive options for this kind of defects. PMID- 15803924 TI - Mechanisms of action of mycophenolate mofetil. AB - Mycophenolate mofetil (MMF, CellCept) is a prodrug of mycophenolic acid (MPA), an inhibitor of inosine-5'-monophosphate dehydrogenase. MPA depletes guanosine nucleotides preferentially in T and B lymphocytes and inhibits their proliferation, thereby suppressing cell-mediated immune responses and antibody formation. MPA also inhibits the glycosylation and expression of adhesion molecules, and the recruitment of lymphocytes and monocytes into sites of inflammation. MPA depletes tetrahydrobiopterin and decreases the production of nitric oxide by inducible NO synthase without affecting the activity of constitutive NO synthases. Activated macrophages produce NO and superoxide, which combine to generate tissue-damaging peroxynitrite. By these two mechanisms MMF exerts anti-inflammatory activity. Unlike calcineurin inhibitors, MMF is not nephrotoxic and does not induce the production of TGFbeta, which is fibrogenic. MMF does not increase blood pressure, cholesterol levels or triglyceride levels in recipients. MMF reduces acute and chronic rejection in allograft recipients and is efficacious in some nephropathies. Evidence is accumulating that MMF may have clinical utility in some autoimmune disorders. PMID- 15803925 TI - Mycophenolate mofetil and systemic lupus erythematosus: an overview. AB - Mycophenolate mofetil (MMF) is an immunosuppressive agent used in transplantation, with evidence of superior protection against acute transplant rejection compared to azathioprine-containing regimens. Subsequently MMF has been used in a variety of autoimmune conditions. The major experience in systemic lupus erythematosus (SLE) has focused on proliferative lupus nephritis. Following its success in the treatment of lupus nephritis, MMF is now being used to control other SLE manifestations such as, lupus disease activity, haematological manifestations and resistant skin lupus. In this review, we discuss our own experience and the literature report about the use of MMF in SLE. PMID- 15803926 TI - Mycophenolate mofetil in animal models of autoimmune disease. AB - Mycofenolate mofetil (MMF-Cellcept) is an immunomodulatory drug utilized extensively in transplant medicine. The efficacy of regimes including Cellcept in preventing allograft rejection, and in the treatment of rejection, is now firmly established. The immunosuppressive actions of this drug enabled the investigation for the beneficial effects in autoimmune diseases. We review the evidence for the contribution of MMF in autoimmunity in animal models of systemic lupus erythematosus (SLE), mercury induced autoimmune glomerulonephritis, diabetes mellitus, experimental autoimmune uveoretinitis, and experimental allergic encephalitis. MMF has an influence on the T and B cell pathways. It is immunosuppressive and anti-inflammatory. PMID- 15803927 TI - Adhesion molecules, mycophenolate mofetil and systemic lupus erythematosus. AB - Mycophenolate mofetil (MMF) has been reproducibly shown to inhibit lymphocyte adhesion and penetration of endothelial cell surfaces. The mechanism is not yet elucidated. In vitro studies on the effects of MMF on cell adhesion molecules (CAM) using human umbilical vein endothelial cells (HUVEC) have shown conflicting results. Different studies have independently shown that MMF increased, decreased or had no effect on intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM-1). Several studies suggest MMF may reduce the endothelial expression of E-selectin. Recent studies have been unable to replicate initial work, which suggested that MMF impaired glycosylation of lymphocyte CAM. The same studies concluded that MMF had no effect on the surface expression of lymphocyte CAM, but altered the binding ability of these molecules. ICAM-1/LFA-1 (lymphocyte function-associated antigen-1), VCAM-1/VLA-4 (very late antigen-4) and P-selectin/PSGL-1 (P-selectin glycoprotein ligand-1) ligand pairs are most likely to be involved. Few in vivo and no conclusive human studies have been carried out. The literature relevant to cell adhesion molecules in systemic lupus erythematosus (SLE) is reviewed in detail. PMID- 15803928 TI - Lupus nephritis: induction therapy. AB - Effective induction therapy is of pivotal importance in minimizing renal parenchymal damage by the active immune-mediated inflammatory processes in severe proliferative lupus nephritis. Preservation of nephron mass is prerequisite to long-term renal survival. Data from US-based studies have shown improved efficacy with induction treatment comprising corticosteroid and cyclophosphamide, compared with corticosteroid treatment alone. Data from European studies have shown similar efficacy with a modified treatment regimen, in which smaller doses of cyclophosphamide were given at weekly or fortnightly intervals over a shortened treatment duration, and the treatment related adverse effects appeared less frequent with the reduced-dose regimen. We have also reported that sequential immunosuppression with prednisolone and oral cyclophosphamide as induction followed by azathioprine maintenance was associated with a high incidence of remission and relatively favourable long-term renal outcome in Chinese patients. However, cyclophosphamide treatment is associated with considerable adverse effects, which could be potentially fatal. Mycophenolate mofetil selectively inhibits lymphocyte proliferation, and thus targets an instrumental step in the pathogenesis of systemic lupus erythematosus. There is accumulating evidence that the combined use of mycophenolate mofetil and corticosteroid presents an effective treatment for severe proliferative lupus nephritis in different ethnic groups, and is associated with much fewer adverse effects compared with cyclophosphamide-based regimens. Recent data from our group also demonstrate the long-term efficacy of mycophenolate mofetil in preserving renal survival, when used continuously as both induction and maintenance therapy. PMID- 15803929 TI - Maintenance therapies for proliferative lupus nephritis: mycophenolate mofetil, azathioprine and intravenous cyclophosphamide. AB - For the treatment of proliferative lupus nephritis, long-term cyclophosphamide (CY) regimens are efficacious, however, at the expense of substantial toxicity. In the last decade, sequential regimens of short-term CY induction followed by either mycophenolate mofetil (MMF) or azathioprine (AZA) maintenance have shown to be efficacious and safe reducing the long-term exposure to CY. In a maintenance study including predominantly Hispanics and African-Americans, the patients who received MMF and AZA maintenance had a higher cumulative probability of remaining free of the composite of death or chronic renal failure (CRF) compared to quarterly intravenous CY (IVCY) maintenance (89% in MMF, 80%, in AZA and 45% in IVCY). Likewise, MMF and AZA maintenance were associated with significantly lower incidence of severe infections (2% in each MMF or AZA, and 25% in IVCY), sustained amenorrhea (6% in MMF, 8% in AZA, and 32% in IVCY), and hospitalizations (one hospital-days per patient-year in each MMF or AZA, and 10 in IVCY). In a European induction study including predominantly Caucasians, patients who received any of two sequential regimens, low dose versus high dose IVCY induction both followed by AZA maintenance, had a high cumulative probability of remaining free of treatment failure (84% in low dose IVCY and 80% in high dose IVCY; treatment failure defined as a composite of free of corticosteroid resistant flare, nephrotic syndrome, doubling creatinine, and persistent elevated creatinine). Low dose IVCY and high dose IVCY induction were associated with low incidence of sustained amenorrhea (4% in each group) and severe infections (11% in low dose and 22% in high dose IVCY induction). Of interest, most of the severe infection episodes occurred while patients were receiving IVCY induction. Finally an Asian study demonstrated that patients with proliferative lupus nephritis could be effectively treated with short-term oral CY induction followed by AZA maintenance. The cumulative probability of complete remission was 76%. The relapse rate was only 11%. The incidence of permanent amenorrhea and infection were 8% and 33%, respectively. None of the Asian patients had an increase in serum creatinine level to double the baseline value. Maintenance therapies with MMF or AZA following short-term CY induction in a sequential regimen are efficacious and safe for the treatment of high-risk patients with proliferative lupus nephritis. PMID- 15803930 TI - Mycophenolate mofetil in nonlupus glomerulonephropathy. AB - Mycophenolate mofetil (MMF) initially found widespread use in the immunoprophylaxis of rejection in organ transplantation. It has subsequently been used in lupus glomerulonephritis, where early studies have shown it to be effective in induction and maintenance therapy. The randomized studies have mostly studied small groups of patients and their conclusions do need to be confirmed in larger studies. MMF has also been used in small numbers of patients in a variety of nonlupus glomerulopathies, which have different underlying immunopathology as well as clinical course, including IgA nephropathy, membranous nephropathy, focal segmental glomerulosclerosis, membranoproliferative glomerulonephritis, hepatitis-C-associated glomerulonephritis and even Goodpasture's syndrome. In this article, we discuss its use in such nonlupus glomerular diseases. PMID- 15803931 TI - Mycophenolate mofetil and neurological diseases. AB - We describe experience with the use of mycophenolate mofetil (MMF) in neurological diseases. Although only small series of patients or case reports were described, MMF is promising in immune-mediated neuromuscular disorders. MMF has been used for the treatment of polymyositis, chronic inflammatory demyelinating polyradiculoneuropathy, and multifocal motor neuropathy. These studies showed that MMF is well tolerated and may be useful in some patients. MMF can be effective alone but mainly as an adjuvant therapy by reducing steroid requirements or the frequency of infusions of IVIg. MMF has also been tested alone as a single drug treatment or in combination with immunomodulatory drugs in multiple sclerosis in open surveillance trials or in phase II studies. None of these studies have been designed to demonstrate a clinical efficacy but preliminary results are very promising. PMID- 15803932 TI - Mycophenolate mofetil and myasthenia gravis. AB - Myasthenia gravis (MG) represents the prototypic autoimmune disorder with well characterized immunopathology. Advances in the diagnosis and treatment of this neuromuscular transmission disorder have significantly improved the management of myasthenic patients. Unfortunately the currently available immunomodulating treatments have significant side effects and some patients do not tolerate them or adequately respond to them. Therefore the possibility of a new immunosuppressant agent that is safe, effective and has steroid-sparing effect is very appealing. Mycophenolate mofetil (MMF) has shown promising effects in MG patients in preliminary studies and is currently being studied in two prospective, randomized, double-blind, placebo controlled, multicenter trials to better establish its role in the treatment of MG. PMID- 15803933 TI - MMF and eye disease. AB - Immunosuppressive treatment has shown to be effective in various ocular inflammatory disorders. Factors limiting their use are the individual response and the rate of side effects. This report summarizes our knowledge about the use of mycophenolate mofetil (MMF) in the treatment of ocular cicatricial pemphigoid (OCP), uveitis, atopic keratoconjunctivitis (AKC), prevention of graft rejection after penetrating keratoplasty (PK) and scleritis. Controlled studies have been performed for prevention of graft rejection after PK, showing MMF as effective in the prevention of graft rejection as cyclosporine A. In experimental uveitis, MMF has been demonstrated to be highly effective in prevention of retinal destruction. A number of studies have now shown that MMF also seems effective in uveitis. There are also studies with smaller patient groups which point out the effectiveness of MMF in OCP, AKC, and scleritis. In most of the studies, the spectrum of side effects was small, compared to other immunosuppressives. PMID- 15803934 TI - Mycophenolate mofetil in systemic vasculitis. AB - The systemic vasculitides are a group of potentially life-threatening multi system autoimmune connective tissue diseases characterized by vascular inflammation. The gold standard therapies include corticosteroids and cyclophosphamide as induction therapy and other agents such as azathioprine and methotrexate to maintain remission. Mycophenolate mofetil (MMF) is increasingly being used in autoimmune disorders and this article reviews the use of this agent in the systemic vasculitides. PMID- 15803935 TI - Mycophenolate mofetil and skin diseases. AB - Mycophenolate mofetil (MMF) is an immunosuppressive drug the efficiency of which has been established in renal transplantation. Recent studies suggest that it may also be effective in the treatment of variant skin diseases especially if the skin lesions are triggered by lymphocytes. Studies have shown efficacy in autoimmune bullous dermatoses, atopic dermatitis and psoriasis. However, there are no placebo-controlled trials that support the use of MMF as first line therapy in these skin diseases. PMID- 15803936 TI - Mycophenolate mofetil in patients with hepatitis C virus infection. AB - Diagnosis and treatment of hepatitis C virus (HCV) -related autoimmune features has become a clinical challenge in HCV-infected patients, in whom chronic liver disease associated with severe autoimmune features may contribute to a very poor prognosis. Both antiviral and immunosuppressive therapies, either alone or in combination, seem likely to have a key role. Based on the experience of mycophenolate mofetil (MMF) use in HCV patients receiving organ transplantation, this new immunosuppressive agent might represent a safe and effective therapeutic option to treat HCV-related extrahepatic features. Recent data are available for the use of MMF in HCV patients with autoimmune manifestations, mainly for autoimmune cytopenias and vasculitic features. MMF may be used as monotherapy or in association with other drugs for cases of HCV-related autoimmune diseases refractory or intolerant to common immunosuppressive treatments, allowing the reduction of the drug dosage and avoiding serious side effects. PMID- 15803937 TI - Where's the money? Spending on physician services rose at a smart pace in 2003, but not across the board. PMID- 15803938 TI - Classifying migraines. PMID- 15803939 TI - More answers to your EHR questions. PMID- 15803940 TI - How to pay for your EHR. PMID- 15803941 TI - EHRs. A way around start-up costs. PMID- 15803942 TI - Going online. The need for speed. PMID- 15803943 TI - The best treatment? It's at your fingertips. PMID- 15803944 TI - Shedding light on shedding pounds. PMID- 15803945 TI - Baghdad, bombs, and a kid with asthma. PMID- 15803946 TI - How to market your practice. PMID- 15803947 TI - Going bare--except for a T-shirt. PMID- 15803948 TI - My rogues' gallery of patients. PMID- 15803949 TI - Returning to practice. PMID- 15803950 TI - Almost 65? What you need to know about Medicare. PMID- 15803951 TI - Healthcare funds. PMID- 15803952 TI - Malpractice. Will a jury think money clouds your judgment? PMID- 15803953 TI - Lawsuit threat? Respond carefully. PMID- 15803954 TI - Accumulation of fenvalerate and related changes in lactate and succinate dehydrogenases activity in functionally different tissues of the freshwater fish, Cirrhinus mrigala (Hamilton). AB - Acute toxicity (LC50) of fenvalerate at 96 h to the freshwater fish Cirrhinus mrigala was 6 microg/L. The accumulation of fenvalerate at different exposure periods (1, 2, 3, 4 d) on different tissues, namely gill, muscle, and liver, was first confirmed by thin layer chromatography and quantified with gas liquid chromatography. Gill accounted for the maximum accumulation of fenvalerate followed by muscle and liver. Enzyme assay indicated an increased activity of lactate dehydrogenase, whereas the activity of succinate dehydrogenase decreased continuously up to 96 h in all selected tissues. The alteration in the activity of these enzymes in the fish treated with fenvalerate might be due to the severe cellular damage leading to the release of these enzymes and impaired carbohydrate and protein metabolism. PMID- 15803955 TI - Acamprosate involvement in triacylglycerol hydrolysis and transacylation with cholesterol in chronically ethanol-drinking rats. AB - Acamprosate (AC) is used as a drug for treating alcoholism. We evaluated the effect of AC on serum triacylglycerol hydrolysis (GEH, glycerol ester hydrolysis), triacylglycerol transacylation with cholesterol (GECAT, glycerol ester:cholesterol acyltransferase), and acylcholesterol hydrolysis (Cease, cholesterol ester hydrolysis) in an experimental model of alcoholism. Ethanol preferring (PRF), non-preferring (NPF), and control (CR) male Wistar rats were treated with AC (500 mg/kg, p.o.) for 21 consecutive days. The beneficial effect of AC on lipid parameters of PRF rats included decreased triacylglycerol, total cholesterol, and LDL-cholesterol, and increased HDL-cholesterol levels. Acamprosate-compensated changes associated with ethanol consumption were observed. Acamprosate treatment decreased GECAT and increased Cease control rats, but increased GECAT and decreased CEase in PRF animals. In all groups of rats, AC treatment did not influence GEH. In conclusion, our results suggest that AC can influence triacylglycerol metabolism by its action on the balance between hydrolysis and transacylation in rats. PMID- 15803956 TI - Link between monthly rates of four subtypes of acute myocardial infarction and their corresponding cosmophysical activity parameters. AB - The aim of this study was (1) to examine the relation between the monthly rate of acute myocardial infarction (AMI) and three cosmophysical activity parameters: solar activity (SA), geomagnetic activity (GMA), and cosmic rays activity (CRA) levels; (2) to study whether different subtypes of AMI: ST-elevation MI (STEMI), non-ST-elevation (NSTEMI), Q-wave (Q-waveMI) and non-Q-wave (NQ-wave MI) are linked with monthly cosmophysical indices or with the daily level (I degrees-IV degrees) of GMA. METHODS: For the first question, we studied for 204 consecutive months (1983-1999) 16,683 patients (including 10405 males) with AMI who were included in the Kaunas Registry and for the second, 3824 AMI patients (2342 males), 72-month data. Cosmophysical data were obtained from the Apatity Neutron Monitoring Station of the Russian Academy of Science. RESULTS: According to neutron monitoring data, total AMI and all its subtypes significantly correlated with monthly levels of CRA and inversely correlated with SA and GMA indices (r = 0.32-0.45; p = 0.0007-0.0001). No significant correlation was found between AMI subtypes and the daily level (I degrees-IV degrees) of GMA. All cosmophysical parameters were stronger in female patients. CONCLUSION: The monthly rates of all subtypes of AMI were significantly correlated with CRA and inversely correlated with SA and GMA, more strongly in female patients. We presume that the environmental factors studied here affect the general patho-physiological components of AMI, and that different subtypes are a consequence of the localization and extent of the process. PMID- 15803957 TI - Studies on the role of ascorbic acid on nickel induced hepatic nucleic acid concentrations in rats. AB - We studied the effect of oral treatment with ascorbic acid (50 mg/100 g body weight) on nickel sulfate-induced (2.0 mg/100 g body weight, i.p.) alteration of nucleic acids and total protein concentration in the liver of Wistar strain male albino rats. Nucleic acids and total protein concentrations in treated rats decreased significantly when compared with untreated controls. The simultaneous administration of ascorbic acid with nickel sulfate resulted in a remarkable improvement of nucleic acids and total protein concentrations in liver in comparison with rats treated with nickel sulfate only. The results indicate that nickel influences the expression of genetic information by reducing hepatic DNA, RNA, and protein concentration in animals. Simultaneous treatment with ascorbic acid was beneficial for fighting against nickel-induced hepatoxicity. PMID- 15803958 TI - Maintenance or interruption of pregnancy by human chorionic gonadotrophin in adrenalectomized albino rats. AB - In the present study, we investigated the efficacy of human chorionic gonadotrophin (HCG) in the maintenance of pregnancy in adrenalectomized rats. Holtzman's strain albino rats were adrenalectomized on day 8 of pregnancy and at the same time were laparotomized to observe the number of implantations. Adrenalectomy on day 8 caused abortions or fetal resorption in almost all rats. Administration of 5 or 50 IU HCG in adrenalectomized rats, from day 8 through 14 or 19, was not able to maintain gestation, resulting in fetal resorption with many placentomas and placental scars. However, a single injection of 5 IU or 1 IU HCG administered on day 8 only maintained the pregnancy to full term in adrenalectomized rats. Hence, the present experiment indicates that replacement therapy of a single dose of 5 IU or 1 IU HCG might be sufficient for maintaining pregnancy in adrenalectomized rats. PMID- 15803959 TI - Evaluation of dimethoate toxicity on pregnancy in albino mice. AB - We investigated whether dimethoate, an organophosphorus insecticide, causes abortion or fetal resorption in pregnant albino mice. Graded doses of 16, 20, 24, and 28 mg/kg body weight/d were administered orally from days 7 to day 15 of pregnancy. Laparotomy was performed on day 8 of pregnancy to note the number of implantations, and the animals were autopsied on day 19. The results revealed no inhibition of pregnancy in all dimethoate treated mice relative to a suitable control group. Treatment with 24 or 28 mg dimethoate caused a significant decrease in the number of implantations, live fetuses, and corpora lutea, but a decreased percent fetal survival and increased percent post-implantation loss and gestation length were not significant when compared with control mice. In all mice treated with 28 mg dimethoate, a significant decrease occurred in the body weight of the ovaries, uterus, and liver when compared with control mice. Following treatment with 16 or 20 mg dimethoate, however, no significant change was found in body and organ weights or in the number of implantations, live fetuses, and corpora lutea, percent post-implantation loss, and fetal survival, or gestation length compared with the corresponding parameters in control mice. The results of this study clearly indicate that dimethoate does not cause abortion or fetal resorption in pregnant mice. A significant decrease in the number of corpora lutea and percent fetal survival observed at higher doses of dimethoate could be due to a toxic effect on the embryo or to a hormonal imbalance. PMID- 15803960 TI - Effect of Scoparia dulcis extract on insulin receptors in streptozotocin induced diabetic rats: studies on insulin binding to erythrocytes. AB - We investigated the insulin-receptor-binding effect of Scoparia dulcis plant extract in streptozotocin (STZ)-induced male Wistar rats, using circulating erythrocytes (ER) as a model system. An aqueous extract of S dulcis plant (SPEt) (200 mg/kg body weight) was administered orally. We measured blood levels of glucose and plasma insulin and the binding of insulin to cell-membrane ER receptors. Glibenclamide was used as standard reference drug. The mean specific binding of insulin to ER was significantly lower in diabetic control rats (DC) (55.0 +/- 2.8%) than in SPEt-treated (70.0 +/- 3.5%)- and glibenclamide-treated (65.0 +/- 3.3%) diabetic rats, resulting in a significant decrease in plasma insulin. Scatchard plot analysis demonstrated that the decrease in insulin binding was accounted for by a lower number of insulin receptor sites per cell in DC rats when compared with SPEt- and glibenclamide-treated rats. High-affinity (Kd1), low-affinity (Kd2), and kinetic analysis revealed an increase in the average receptor affinity in ER from SPEt and glibenclamide treated diabetic rats having 2.5 +/- 0.15 x 10(10) M(-1) (Kd1); 17.0 +/- 1.0 x 10(-8) M(-1) (Kd2), and 2.0 +/- 0.1 x 10(-10) M(-1) (Kd1); 12.3 +/- 0.9 x 10(-8) M(-1) (Kd2) compared with 1.0 +/- 0.08 x 10(-10) M(-1) (Kd1); 2.7 +/- 0.25 x 10(-8) M(-1) (Kd2) in DC rats. The results suggest an acute alteration in the number of insulin receptors on ER membranes in STZ-induced diabetic rats. Treatment with SPEt and glibenclamide significantly improved specific insulin binding, with receptor number and affinity binding (p < 0.001) reaching almost normal non-diabetic levels. The data presented here show that SPEt and glibenclamide increase total ER membrane insulin binding sites with a concomitant significant increase in plasma insulin. PMID- 15803961 TI - Morphometric analysis of follicular growth and biochemical constituents in albino rats exposed to mancozeb. AB - Mancozeb, an ethylenebisdithiocarbamate (EBDC), was administered orally at a dose of 700 mg/kg body weight/day to female virgin rats for 5, 10, 20, and 30 days to examine the effect on ovarian follicular development. No significant change occurred in the number of estrous cycles and the duration of proestrus, estrus, and metestrus, but mancozeb treatment for 5 days significantly increased the duration of diestrus. Mancozeb treatment for 10 days significantly increased the number of estrous cycle and the duration of estrus, with a concomitant significant increase in diestrus, but no change in proestrus and metestrus. With mancozeb treatment for 20 and 30 days, the number of estrous cycles and duration of proestrus, estrus, and metestrus were significantly decreased, with a concomitant significant increase in the duration of diestrus. Exposure of rats to mancozeb for 5 days resulted in a significant decrease in stage II and the total number of healthy follicles but no change in atretic follicles. Mancozeb treatment for 10 days resulted in a significant decrease in stages I, II, and IV and in the total number of healthy follicles. A significant increase in atretic follicles was found in rats treated with mancozeb for 20 and 30 days. No significant change occurred in body and organ weights in any group, but the thyroid weight of 20 and 30 days mancozeb-treated rats was significantly increased. The level of protein in the ovary was significantly decreased, but no change was found in the uterus and liver of mancozeb-treated animals. The level of glycogen was significantly decreased in the ovary and the uterus with mancozeb treatment, but not in the liver. With mancozeb treatment, the levels of phospholipids and neutral lipids were significantly increased in the liver but significantly decreased in the uterus. The change in the biochemical constituents of ovary, uterus, and liver was duration dependent. The results of the study thus indicate a marked disruption of the estrous cycle, pathological changes in the gonads, and organ-specific biochemical changes in rats after exposure to mancozeb. PMID- 15803962 TI - Dose-dependent serum biochemical alterations in Wistar albino rats after Palamneus gravimanus (Indian black scorpion) envenomation. AB - Palamneus gravimanus envenomated rats showed dose-dependent alterations in serum biochemical parameters. Sub lethal doses of 100, 200, and 400 microg/kg of P. gravimanus venom were injected intramuscularly into rats. Blood samples were collected by heart puncture before and 4 h after crude venom administration. Serum was analyzed for glucose, blood urea nitrogen (BUN), uric acid, total protein, cholesterol, sodium, potassium, inorganic phosphorus, alkaline phosphatase, aspartate aminotransferase (AST-SGOT), alanine amino-transferase (ALT-SGPT), lactate dehydrogenase (LDH), and creatinine phosphokinase (CPK). Statistically significant increases in serum levels of glucose, creatinine, AST, ALT, BUN, CPK, and LDH and significant decreases in serum levels of total protein, uric acid, cholesterol, calcium, and potassium 4 h after venom administration could be due to the toxic action of P. gravimanus venom on certain organs in rats. PMID- 15803963 TI - Biochemical and clinicopathological changes induced by Bungarus coeruleus venom in a rat model. AB - Bungarus coeruleus, a common venomous snake allied to the cobra, is responsible for most envenomations in India. This study examines the pathological effects of B. caeruleus venom and the associated biochemical changes in a rat model. Increased serum aspartate aminotransferase (48%), creatine kinase (30%), and lactate dehydrogenase (6%) were detected after a sublethal dose of 25 microg/kg of B. caeruleus venom was injected intramuscularly into rats observed for 180 min. The venom induced hyperglycemia and increased serum alkaline phosphatase (55%) and urea (90%) concentrations, whereas cholesterol and triglycerides remained normal. Histopathological changes in the heart-hemorrhage, multifocal areas of myocardial fiber necrosis-and constriction of blood vessels in the kidney, with congested vessels, hemorrhage and necrosis of proximal tubules, liver congestion, and hemorrhage were found. PMID- 15803964 TI - Modulation by 4-aminopyridine and calcium on the effect of adenosine on isolated spontaneously beating rat atria. AB - AIM: The functional role of Ca2+ in pacemaker depolarization and in contraction was studied by challenging the inhibitory actions of adenosine to 4-AP and Ca2+ in the spontaneously beating rat atria. EXPERIMENTAL: Studies were performed in adenosine-treated (0.3-100 microg/ml), adenosine + 4-aminopyridine (4-AP) (4mM) treated, and Ca2+ (3.6mM or 400 microg/ml)-treated groups. All values were expressed as mean +/- S.E.M. Statistical analysis was done using Students t-test, where p < 0.05 was considered significant. RESULTS: Adenosine (0.3-100 microg/ml) dose-dependently depressed both the beating frequency and contractility, more powerfully on the latter. 4-AP (4 mM) and Ca2+ (3.6 mM or 400 microg/ml) respectively increased force of contraction (203.4% and 199.3%) and spontaneous beating frequency (104.9% and 111.3%). 4-AP and Ca2+ significantly antagonized the inhibitory actions of adenosine on cardiac processes, more strongly on contractility and with 4-AP more potent than Ca2+. CONCLUSION: (1) cardiac contractility was more responsive than pacemaker activity to the inhibitory action of adenosine, (2) 4-AP and Ca2+ acted to antagonize cardiac inhibition induced by adenosine, (3) the potency of antagonism appeared to be higher with 4 AP, and (4) the antagonism of adenosine by 4-AP on beating frequency is stronger with respect to contractility and is variable with Ca2+. PMID- 15803965 TI - Sexual arousal in sexually sluggish old male rats after oral administration of Eurycoma longifolia Jack. AB - Eurycoma longifolia Jack commonly known as Tongkat Ali in Malaysia, has been used in Malaysia to increase male virility and sexual prowess. The objective of this study is to evaluate sexual arousal in sexually sluggish old male rats, 24 months old and retired breeders, receiving 200, 400, or 800 mg/kg of various fractions of E. longifolia Jack, twice daily, for 10 days. Control rats received 3 ml/kg of normal saline. The aphrodisiac effect was monitored by the act of yawning and stretching because yawning, either alone or associated with stretching, is considered an ancestral vestige surviving throughout evolution that promotes sexual arousal. The results showed that 800 mg/kg of E. longifolia Jack increased yawning by 50% and stretching by 16.7% in sexually sluggish old male rats, by 676 719% and 31-336%, respectively, in sexually active male rats, and by 22-44% and 75-100%, respectively, in middle aged, 9 months old and retired breeders. We conclude that the results of this study support the folk use of this plant as an aphrodisiac. PMID- 15803966 TI - [Emergency medicine]. PMID- 15803967 TI - A junctional rhythm with retrograde conduction and bradycardia as a dysrhythmia caused by oral phenytoin toxicity. PMID- 15803968 TI - Myocarditis, an adolescent presentation: case report and review of literature. PMID- 15803969 TI - Portuguese man of war: case report. PMID- 15803970 TI - [Resuscitation carts: how to organize and keep uniformity in drug supply]. PMID- 15803971 TI - Implantable electrical devices for dysrhythmia: what every emergency physician must know. PMID- 15803972 TI - Most common injuries associated to scooters seen in the emergency department. AB - The foot propelled scooter is a popular toy preferred by girls and boys, but at the same time, these put the children at risk of some kind of injury. The literature had reported that the fractures at the upper extremities are the most frequent injury observed related to the use of a scooter. The purpose of our study was to describe the injuries related to the scooters use in a sample of injuries handled at the UPR Hospital at Carolina, Puerto Rico. A total of 62 patients who arrived to the ED and were evaluated for some kind of injury related to the use of scooters were included in our study. Fractures (32.3%) constituted the most frequent injury observed in the sample, followed by the lacerations (29.0%), and the abrasions (11.3%). The parents, doctors and the healthcare workers should be made aware of the causes and prevention of injuries caused by the foot propelled scooters, because this situation is potentially preventable. PMID- 15803973 TI - [Emergency medicine curriculum of the University of Puerto Rico. History and contribution to health care in Puerto Rico]. PMID- 15803974 TI - [The rescue of Ramon Power]. PMID- 15803975 TI - [Puerto Rico needs more primary physicians]. PMID- 15803976 TI - [Family practice]. PMID- 15803977 TI - [In addition, demanding fast, accessible, sustainable, and efficient services]. PMID- 15803978 TI - [Training for family physicians in the management of advanced disease and death]. PMID- 15803980 TI - [Factors that encourage or discourage the practice of breastfeeding. Comparison of trends with the 1998 study and comparison of pro-breastfeeding group versus traditional group]. PMID- 15803979 TI - [Compliance with current therapy guidelines in hypertensive patients at an emergency service]. AB - Arterial hypertension is one of the most prevalent health problems and one of the most important risk factors for cardiovascular and cerebrovascular disease. The purpose of this study was to identify if hypertensive patients are being treated in accordance to current recommendations (JNC-7) for this condition and to estimate the level of blood pressure control in the subjects at the moment of their participation in the study. A total of 138 patents with previous diagnosis of arterial hypertension were recruited from visitors to the Emergency Room at Dr. Pila Hospital in Ponce, Puerto Rico during the months of November 2003 through April 2004. Data was collected in the form of a questionnaire, administered by interview. The questionnaire included demographic data, blood pressure level, anthropometric variables, reported history of chronic diseases, use of antihypertensive agents and other variables. At the moment of their participation, 77% of the subjects had uncontrolled hypertension. Only 25% of subjects reported present use of diuretic agents, which are highly recommended as first line treatment for hypertension. The majority (54%) of subjects were being treated with only one antihypertensive agent. The majority (58%) of patients were not being treated in accordance with current recommendations (JNC-7) under the category of "compelling indications". The results of this study could very well indicate tendencies and patterns of treatment in hypertensive patients of our community. This may warrant further research and inquiry for explanations. PMID- 15803981 TI - [Medical errors in the continuous improvement of quality]. AB - The increase in medical errors during the last four decades of past centuries can be attributed to several causes: the human fallibility always present, concern in quality and continuity of physician-patient relationship and to the increased advance in technology innovation. These errors attract the attention of Agencies and Organizations involved in Healthcare Quality. From the work of these a series of recommendations were issued for the prevention of medical errors. The action level is a systemic one, due to the characteristic of the healthcare services. PMID- 15803982 TI - [Spiritual medicine]. PMID- 15803983 TI - [Alzheimer's disease: new treatment options]. PMID- 15803984 TI - [The project on the future of family practice: a Puerto Rican perspective]. PMID- 15803985 TI - Risk and protective factors associated with youth violence among secondary school students in a nationally representative sample in Puerto Rico. AB - During the past decade, youth violence has received increasing attention as a major public health issue in Puerto Rico as well as in the United States. This study sought to identify risk and protective factors of youth violence in a representative sample of school adolescents in Puerto Rico. Risk and protective factors were grouped into five domains: individual, family, peer group, school and community. From a total of 2,385 participants, 10.7% reported at least one violent behavior and 3.4% reported two or more violent behaviors. In multiple regression analysis the risk factors identified were male gender, junior grade students, having a favorable attitude towards antisocial behavior, use of ecstasy, involvement with antisocial peers and reporting antisocial parents. Participation in family decisions was the only protective factor associated with violence. Findings from this study could have important implications for the development of preventive programs for the adolescent population in Puerto Rico. PMID- 15803986 TI - [Puerto Rican youth at risk]. PMID- 15803987 TI - Typical and atypical presentations of gastroesophageal reflux disease and its management. AB - Gastroesophageal Reflux disease (GERD) has a common clinical presentation of a burning discomfort in the retroesternal area, regurgitation and dysphagia. Yet, an estimated of 20 to 60 percent of patients with GERD have head and neck symptoms without any appreciable heartburn. Careful history and a meticulous physical exam can guide us to have a correct diagnosis and give adequate treatment. Other methods, such as gastroscopy and gastric pH monitoring, as well as other diagnostic studies can help us to confirm the diagnosis. Once we have the correct diagnosis stabilized, life style modification should be the first step in the management of GERD, aided with antacids, H2 receptors antagonists and/or Proton pump inhibitors. Family physicians should be aware that helping patients to understand the cause of their symptoms and reinforcing the life style modifications will bring better control of the disease and patients can have improvement of their symptoms leading to possible cure of the disease. Gastroesophageal Reflux Disease (GERD) is defined as the movement of gastric contents into the esophagus without presence of vomiting. It is frequently associated with heartburn, the sensation of burning discomfort in retrosternal area, that moves upward, toward the throat. GERD is a chronic, relapsing condition with associated morbidity and adverse impact on quality of life. The purpose of this article is to give an overall look at the clinical presentations of GERD with typical and atypical symptoms, the various presentations of this disease in all of the age groups, and to identify all of the aspects that contribute to the progression and solution of this problem. PMID- 15803988 TI - [History of Family Practice in Puerto Rico]. PMID- 15803989 TI - [One serves serving]. PMID- 15803990 TI - [Severo Ochoa de Albornoz y Perez, Physiology and Medicine Nobel Prize and his relationship with Puerto Rico]. PMID- 15803991 TI - [The burial. Oil painting by Francisco Oller, Museum of the University of Puerto Rico]. PMID- 15803992 TI - [Danish clinical intervention research--can it be done better?]. PMID- 15803993 TI - [Calcium homeostasis and bone turnover in pregnancy and breast feeding]. PMID- 15803994 TI - [Pharmacological treatment of osteitis deformans (Paget's bone disease)]. PMID- 15803995 TI - [Heredity and osteoporosis]. PMID- 15803996 TI - [Physical activity, smoking and the significance of vitamins A and D in osteoporosis]. PMID- 15803997 TI - [Secondary osteoporosis, epidemiology and risk factors]. PMID- 15803998 TI - [Drugs with side effects affecting calcium homeostasis, bone metabolism and risk of fractures]. PMID- 15803999 TI - [Evaluation of the effects of treatment of osteoporosis]. PMID- 15804000 TI - [Development of the methodology of therapeutic trials. A study of papers published in Ugeskrift for Laeger during the period 1910-2000]. PMID- 15804001 TI - [Changes in bone mineral density in the proximal femur 18 months after total hip arthroplasty with Exeter stem]. PMID- 15804002 TI - [Children undergoing chronic hemodialysis. Results from Rigshospitalet, 1991- 2002]. PMID- 15804003 TI - [Preventive health examination of children--how are they done? A descriptive study of the practical handling]. PMID- 15804004 TI - [Green amniotic fluid--should suction be used?]. PMID- 15804005 TI - [Occurrence and diagnosis of bacterial tick-borne diseases]. PMID- 15804006 TI - [Referring to MEDLINE indexing]. PMID- 15804007 TI - [Mammography screening astray]. PMID- 15804008 TI - [Observing the distribution of Annexin I and cPLA2 in the palatal process of mice by immunohistochemical staining]. AB - OBJECTIVE: To observe the distribution of Annexin I and cytosolic phospholipose A2 (cPLA2) in the palatal process of dexamethasome teratogenerated mice. METHODS: Cutting along the coronary plane of the heads of fetal mice to obtain palate in the 14th, 15th, 16th day of gestation by 10-week-old in-bred mice. The distribution of Annexin I and cPLA2 was checked by immunohistochemical staining. RESULTS: During the fusing of the palatal processes, the staining of Annexin I and cPLA2 was positive in the epithelial and mesenchymal cells of the palatal process, and the intensity of staining changed with time. CONCLUSION: Annexin I and cPLA2 can modulate the development of fetal palate to some extent, and they may be important mediators in the development of cleft palate induced by dexamethasone. PMID- 15804009 TI - [The distribution of collagen I, II, X and alkaline phosphatase in the development of condylar cartilage of fetal mouse mandible]. AB - OBJECTIVE: The purpose of this study was to investigate the distribution of collagen I, II , X, alkaline phosphatase (ALP) and their roles during initiation of condylar cartilage of the fetal mouse. METHODS: Coronary sections of mandible of mouse embryo aged from 14th to 18th day were studied under light microscope after stained by immunohistochemical method with antibody of types I, II, X collagen and ALP. RESULTS: On the 14th day of mouse embryo, it was found that mesenchymal cells condensation continuous with the periosteum. Type I collagen and ALP were positive behind the terminal of the ossifying mandibular periosteum where future condylar will form. On the 15th day, positive staining for types I, II collagen was found in mesenchymal cells around hypertrophic cells and type X collagen was detected in hypertrophic cells. ALP was positive in both mesenchymal cells and hypertrophic cells. On the 16th day, type I collagen was observed from periosteal osteogenic cells and mesenchymal cells of the fibrous cell layer to the upper hypertrophic cell layer while Type II collagen was restricted from the lower polymorphic cell layer to the bottom of the hypertropic cell layer. Type X collagen was positive in the hypertrophic cell layer. ALP was positive in periosteal osteogenic cells and hypertrophic chondral cells, but not in the polymorphic cell layer. CONCLUSION: Development of condylar cartilage is different from that of limb bone. Types I, II, X collagen are expressed in the condylar chondrocyte on the early stage of endochondral ossification. The histology evidence supports the conjecture that condylar cartilage is derived from differentiated mesenchymal cells of the preperiosteum or periosteum of the mandible where ALP is positively expressed. PMID- 15804010 TI - [Effects of different TiNi-SMA distractor on the height of alveolar ridge augmentation]. AB - OBJECTIVE: To discuss the effects of titanium nickel shape memory alloy (TiNi SMA) distractor with different stretching force and different distance between two fixed ends on the height of alveolar ridge augmentation. METHODS: The mini distractors were all made of TiNi-SMA wire. The maximum stretching force and the distance between two fixed ends according to different distractors (A, B and C) were 20 mm, 700 g; 15 mm, 1000 g; 20 mm, 1000 g. 15 adult mongrel canines were divided into three groups randomly and extracted the mandibular premolars and first molar. One month later, box-shaped osteotomies of the alveolar bone were carried out in the left mandible and the TiNi-SMA distractors were buried. The bone height was measured and the radiographs were taken before and five weeks post operation continuously. RESULTS: One week post operation, the vertical ridge augmentation of group A, B and C averaged (7.55 +/- 1.02) mm, (4.97 +/- 0.34) mm, (8.54 +/- 0.82) mm and the height remained (7.20 +/- 1.05) mm, (4.93 +/- 0.21) mm, (8.04 +/- 0.78) mm five weeks post operation. There was no statistic difference in the height of alveolar ridge augmentation between Group A and C, but Group B showed significant difference with the other two groups. In each group, the height did not changed significantly with the time. The radiographs showed obvious distraction gap post operation, and the new bone regenerated five weeks later. CONCLUSION: It suggests that in the scope of suitable stretching force, the height of alveolar ridge augmentation was determined by the size of TiNi-SMA distractor, that is to say, determined by the distance between the two fixed ends. PMID- 15804011 TI - [The design of Co-Cr-Mo alloy combining the framework with porcelain fused to metal restorations and determination of the mechanical properties]. AB - OBJECTIVE: To investigate a kind of Co-Cr-Mo alloys used for both porcelain fused to metal (PFM) restorations and casting framework of removable partial dentures. METHODS: The Co-Cr-Mo alloy underwent the design for elementary compositions of the alloys and the production from the raw materials by means of a vacuum melt furnace. The strength, hardness, plasticity and casting ability of the alloy were examined with metal tensile test. Vickers hardness test and grid casting were examined respectively. The microstructure of the Co-Cr-Mo alloy was also inspected by scanning electron microscope and X-ray diffraction analysis. RESULTS: The elementary composition of DA9-4 alloy mainly consisted of Co 54% 67%, Cr 21%-26%, Mo 5%-8%, W 5%-8%, Si 1%-3%, Mn 0.1%-0.25% and trace elements. The yield strength of the alloy was 584 MPa, while the tensile strength was 736 MPa. The coefficient of expansion was 15.0%, the Vickers hardness reached 322, and the casting ratio exibited 100%. CONCLUSION: The DA9-4 Co-Cr-Mo alloy used for PFM and framework shown in this paper can meet the clinical demands and have reached the objects of the experiment plan. PMID- 15804012 TI - [Effect of two different ring liner thickness on the precision of castings]. AB - OBJECTIVE: To investigate the effect of two different ring liner thickness on the deformation of resulted castings. METHODS: GC No.2 casting rings (50 mm in height and 39.5 mm in diameter) were lined with two dry ceramic fiber liners with thickness of 0.70 mm and 0.45 mm. Plate-like wax patterns with pre-marked points were invested with a gypsum-bonded investment and the rings were cast with a silver-based dental alloy. The difference between the castings and the wax patterns at corresponding points was calculated to represent the dimensional change of the investment in the rings at different directions. RESULTS: The expansion of the investment in the ring was non-isotropic, with larger radial direction expansion than the axial direction expansion (P < 0.01). In the scope of this study, the effect of two different ring liner thickness on the casting precision was statistically non-significant (P > 0.05). CONCLUSION: The liners with two different thickness failed to eliminate non-isotropic deformation of the castings under the testing conditions of this study. PMID- 15804013 TI - [Cloning and sequence analysis of recombinant fusion gene of Escherichia coli heat-liable enterotoxin B subunit and Actinobacillus actinomycetemcomitans fimbria associative protein]. AB - OBJECTIVE: To clone the recombinant fusion gene of Escherichia coli heat-liable enterotoxin B subunit (Ltb) and Actinobacillus actinomycetemcomitans fimbria associative protein (Fap). METHODS: Two couples of primers were designed for PCR according to the known sequence of ltb and fap. The ltb and fap gene were obtained by amplification PCR technique from plasmid EWD299 of Escherichia coli and Actinobacillus actinomycetemcomitans 310 DNA respectively, and fused them by PCR. The fusion gene ltb-fap were cloning into plasmid pET28a(+). The recombined plasmid pET28a ltb-fap was transformed into Escherichia coli DH5alpha. The recombinant was screened and identified by restriction enzyme and PCR. The cloned gene was sequenced. RESULTS: The ltb-fap about 531bp in size was obtained successfully, and identified by PCR, restrictive enzyme and sequence analysis. CONCLUSION: The vector of pET28a ltb-fap was obtained. PMID- 15804014 TI - [MGB probes detect Streptococcus mutans and Streptococcus sobrinus in real time]. AB - OBJECTIVE: To detect and distinguish Streptococcus mutans (S. mutans) and Streptococcus sobrinus (S. sobrinus) quickly in epidemiology and investigate the distribution of S. mutans in the oral of children with rampant caries. METHODS: Designed minor groove binder (MGB) probes according to the gtf gene of S. mutans and S. sobrinus. Detected 9 reference strains of Streptococcus mutans group by MGB probes in real time and after cultivation. Evaluated the results of these two methods. 92 dental plaques from pre-school children with rampant caries were detected in real time with MGB probes. RESULTS: The primers could amplify the target sequences specificity and distinguished S. mutans and S. sobrinus from each other using MGB probes. Though the fluorescence occurred earlier in S. mutans than in S. sobrinus, they had the same results in nature. In 92 children with rampant caries, the detective ratio of S. mutans was 96.7% and that of S. sobrinus was 32.6%. All the samples which could detect S. sobrinus were positive for S. mutans. CONCLUSION: The primers and probe designed from gtf genes of S. mutans and S. sobrinus can amplify the target sequence and distinguish them from each other in real time. PMID- 15804015 TI - [Evaluation of mastication function after three-dimensional maxillary reconstruction]. AB - OBJECTIVE: To evaluate the mastication function of the patient whose maxilla was reconstructed with individual titanium mesh and Chinese flap or combined with fibular flap. The superiority of this method on maxillary functional reconstruction was testified. METHODS: Since March of 2001, 10 cases of maxillary defect were reconstructed with individual titanium mesh and Chinese flap or combined with fibular flap, and routine removal partial dentures were fixed after 3 - 6 months postoperatively. Assessment of occlusal force was proceeded by T Scan II system (Tekscan company, USA). RESULTS: The occlusal force analysis results indicated the asymmetry index of bite force and asymmetry index of occlusal contact area differed significantly between preoperation and postopertion (P < 0.05). The recovery rate of total occlusal force was between 27.05%-74.06%, and the average was 50.15%. CONCLUSION: This new three-dimensional maxillary functional reconstruction method had a satisfactory recovery in both of contour and function recently, especially restored the mastication function effectively. PMID- 15804016 TI - [The clinic skill in fixed appliance based on characteristics of Chinese normal occlusion]. AB - OBJECTIVE: To study the bracket placement and arch wire bending based on ethnic differences and individual differences of normal occlusion. METHODS: The prominence, tip, torque, upper first molar offset of crown and arch form between Chinese and Caucasian normal occlusion were compared. RESULTS: The results showed the ethnic differences of prominence, tip, torque, upper first molar offset of crown and arch form between Chinese and Caucasian normal occlusion. The placement of bracket was influenced by the crown morphology. CONCLUSION: The adjustments of the bracket placement and arch wire bending with Edgewise and pre-adjusted appliance are necessary to adapt to ethnic difference and individual difference. PMID- 15804017 TI - [Cephalometric evaluation of the effects of pendulum appliance on various vertical growth patterns]. AB - OBJECTIVE: The aim of this study was to investigate the effects of the pendulum appliance in treating dental Class II patients with various vertical growth patterns. METHODS: The samples (n = 30) were divided into three groups equally based on their FMA. Pretreatment and post-treatment cephalometric radiographs were taken to measure the changes. RESULTS: The amount of upper molar distalization in the low-angle group was the fewest, and that in the high-angle group was the most. Upper molars had been intruded insignificantly. The amount of anchorage loss at the first premolars and overjet increased at incisors was different in the three groups. The biggest change happened in the low-angle group, and the smallest in the high-angle group. CONCLUSION: The results of this study showed that pendulum appliance could move the upper molars distally in a short period of time. The upper molars in different groups were intruded insignificantly. Pendulum appliance could be used to move the upper molars distally in high-angle cases. PMID- 15804018 TI - [A study of 202 periodontitis subjects in Chengdu]. AB - OBJECTIVE: To study the severity of periodontitis and risk factors in Chengdu. METHODS: 202 periodontitis patients (65 male, 137 female), aged from 25 to 60, were requested to fill a questionnaire. Probing depth (PD), clinical attachment level (CAL), gingival recession and bleeding on probing (BOP) on 6 sites of each tooth were measured and recorded. RESULTS: The mean PD, AL, gingival recession and BOP% of 202 subjects was (3.2 +/- 0.31) mm, (3.5 +/- 0.37) mm, (0.3 +/- 0.02) mm and 21.16%. 59% of subjects missed at least one tooth. 129 subjects suffered with initial to moderate periodontitis. 73 subject suffered with advanced periodontitis. 40, 86, 55 and 21 subjects had received college education, high school education, middle school education and primary school education. 18% of subjects had smoking history, 67% subjects had tea/coffee history, 66% of subjects had psychosocial problem, and only 8% of subjects had received regular periodontal treatment. There is no relationship between the severity of periodontitis and education. CONCLUSION: It is very important to develop an education program on oral healthy for people in Chengdu. PMID- 15804019 TI - [Measurement and analysis of the intra-articular pressure in temporomandibular joint with sudden-onset, severe closed lock]. AB - OBJECTIVE: The study aims to find out the fluctuating curve of the intra articular pressure in temporomandibular joint with sudden-onset, persistent, severe closed lock and discuss the mechanism of its formation. We also investigate the effects of the arthrocentesis. METHODS: 20 affected sides in 16 patients were collected. A No. 8 syringe needle was used to pierce into the upper compartment of TMJ. The pre-arthrocentesis intra-articular pressure was measured by the pressure transducer via the flexural rigid tubing at open and close bite. The curve and mean value were documented during the operation. RESULTS: The patients with sudden-onset, persistent, severe closed lock had significantly low negative intra-articular pressure in their affected temporomandible joints. The average pressures was (-9.947 +/- 8.854) kPa at open bite and (-6.475 +/- 4.147) kPa at close bite. CONCLUSION: The TMJs with sudden-onset, persistene, severe closed lock has particular characters on etiology and clinical behavior. Arthrocentesis is one of the effective treatments to the diseases. PMID- 15804020 TI - [The correlation between personal anxiety and dental fear among 1975 junior college students in southwest China]. AB - OBJECTIVE: To reveal the correlation between personal anxiety (PA) and dental fear (DF) among the junior college students in Southwest China. METHODS: The Corah's questionnaire and personal anxiety measuring indicator (SAS) were conducted on 1 975 junior college students in Chengdu. The collected data was analyzed by Software SPSS 10. RESULTS: A correlation was found between DF and PA (chi2 = 3.97, P < 0.05). The score of DF was well related with the score of the 15 questions designed (r = 0.913, P = 0. 000). The side affections during dental treatment process had significant effects on DF (r = 0. 409, P = 0.000). CONCLUSION: A correlation was found between DF and PA among the junior college students in Southwest China. PMID- 15804021 TI - [Oral health knowledge, attitude and behavior in adults of Deyang city, PR China]. AB - OBJECTIVE: To investigate the oral health knowledge, attitudes and behavior of urban, rural and suburban adults in Deyang, PR China, and analyze the influence of several variables on the utilization of dental services. METHODS: 1010 adults with 35-44 years old from urban, rural and suburban areas were interviewed by Questionnaire for Adults, WHO Integrated Package for Basic Oral Care. RESULTS: More than half of the surveyed adults claimed "having pain of mouth during the last 1 year". More urban adults (61.3%) used fluoridate toothpaste than suburban and rural adults (6.9%, 16.8%). 61.0% and 71.5% of rural and suburban adults had never seen a dentist, compared with 32.0% of urban adults. Logistic regression analysis showed that urban adults having pain in mouth during the last year were more likely to visit a dentist in the last 2 years. CONCLUSION: Although the oral health attitude is good among the people in Deyang, the knowledge is poor. Dental service utilization was not enough and the situation was also worse for rural adults. PMID- 15804022 TI - [The histological research between the mode of invasion and cervical lymph node metastasis in squamous cell carcinoma of the oral cavity]. AB - OBJECTIVE: The relationship between the mode of tumor invasion in the tumor-host borderline and the frequency of cervical lymph node metastasis was investigated in squamous cell carcinoma of the oral cavity. METHODS: 200 cases with histologically proven squamous cell carcinoma of the oral cavity were studied by histological method with HE stained. The mode of invasion in the tumor-host relationship was classified into five grades by Yamamoto's criteria. RESULTS: With regard to the relationship between the mode of invasion and metastasis, the more invasive the tumor tissue was, the more frequent the metastasis formed (P < 0.001). The frequency of metastasis in grades 1 and 2 was low (0 and 5.9%, respectively), The frequency of metastasis in grades 3 was moderate (14.3%), and that in grades 4c and 4d was highly rapid (63.0% and 82.9%, respectively). Single node metastases were frequent in grade 3 and grade 4c (66.7% and 58.8%, respectively), while plural node metastases were frequent in grade 4d (70.6%, P < 0.05). Moreover, the distribution of metastasized lymph nodes was focused on level 1 (41.2%) or level 1 and 2 (79.4%) in grade 4c and was dispersed from level 1 to 4 in grade 4d (P < 0.05). In the present study, the degree of differentiation did not correlate well with the frequency of metastasis. CONCLUSION: These results indicate that the more invasive the tumor cells were to the host, the more frequent the metastasis formed. The different mode of invasion would accompany with different frequency of metastasis, different number and distribution of metastasized lymph nodes. PMID- 15804023 TI - [A retrospective study on the complications after modified parotidectomy in benign tumors of parotid gland]. AB - OBJECTIVE: A retrospective clinic study is presented of surgical patients undergoing a modified parotidectomy with conserving the sub-superficial musculoaponeurotic system (sub-SMAS) and great auricular nerve and reconstructing by the stemocleidomastoid flap (SF). The incidence of clinical complications was observed after the parotidectomy. METHODS: 226 patients were divided randomly into four groups. Group 1 (94 cases) were operated by the conserving the sub SMAS. Group 2 were operated by the sub-SMAS and constructed by SF. Group 3 (57 cases) were treated by the subcutaneous flap (sub-CF). Group 4 (33 cases) were received by sub-CF and SF. The surgical technique was described and the incidence of clinical Frey's syndrome was evaluated by minor test (starch-iodine test) and observed gustatory sweating syndrome. The feeling and concave of the region of auricular lobule were followed up. RESULTS: The incidence of Frey's syndrome in group 1 (5.32%) or group 2 (4.76%) was significantly lower than that of group 3 (49.1%) or group 4 (51.5%) through by subjective observation and minor test (P < 0.005). The significance between group with SF and group without SF existed in protecting the concave of the region of auricular lobule (P < 0.05). The feeling of the region of auricular lobule had no change by preserving the great auricular nerve. CONCLUSION: There was good effect to prevent gustatory sweating syndrome in parotidectomy of reserve parotid fascia. The aesthetics and sense of the region of auricular lobule could be prevented by using SF and reserving the great auricular nerve. PMID- 15804024 TI - [Brain activities during maximum voluntary clenching with soft occlusal pad in healthy adults by functional magnetic resonance imaging]. AB - OBJECTIVE: Using functional magnetic resonance imaging (fMRI) to detect the cerebral cortical somatotopy during maximum voluntary clenching with soft occlusal pad in healthy adults. By comparing with the cerebral cortical activation during clenching with and without soft occlusal pad we try to detect the central mechanism of the rule of the occlusal pad. METHODS: Ten healthy subjects were selected and scanned by Elscint/GE 2.0 Tesla MR system. Block design was used and the movement pattern was maximum voluntary clenching with soft occlusal pad. The fMRI data were analyzed by SPM99 software and group map analysis was done. RESULTS: The activation areas were found in bilateral or unilateral motor cortex, somatosensory cortex, prefrontal cortex, temporal cortex, premotor cortex, insula, frontal operculum, basal ganglia (putamen), parietal cortex and cingulate. Group map analysis showed activation in bilateral motor cortex, right somatosensory cortex, bilateral basal ganglia, and bilateral insula. The activation of left motor cortex was significantly higher than right side. Compared with cortical activation without occlusal pad, the motor and somatosensory cortex changed a lot. CONCLUSION: The changes of the activation of motor and somatosensory cortex during clenching after wearing the soft occlusal pad might be the central mechanism of the rule of the occlusal pad. PMID- 15804025 TI - [A study on the chondrogenesis of the compound of alginate gelatin and bone marrow stromal cells in vivo]. AB - OBJECTIVE: To investigate the chondrogenisis by alginate gelatin and rats' bone marrow stromal cells (BMSCs) chondrogenicly induced in vitro. METHODS: Thirty-two male adult SD rats were assigned randomly to experimental and control groups. In experimental group, bone marrow was obtained from the right tibias of all the rats. After expanding and culturing 3 passages, induced BMSCs by chondrogenic culture medium for 10 days. Suspended induced cells in alginate gelatin, and injected the complex into the hypodermic tissue of the backs of rats autogenously. In control group only alginate gelatins were injected. The grafts were taken out for examinations 4 and 8 weeks after the operations. RESULTS: Considerable cartilage appeared in experimental group 8 weeks after operations. Regular HE staining and alcian blue staining showed a great deal of cartilage holding chondrocyte masses surrounded by abundant matrix. Alginate gelatin decompounded obviously, and the rest distributed among newly formed cartilage. No cartilage appeared in control group all through. CONCLUSION: BMSCs and alginate gelatin have a beautiful future in cartilage tissue engineering. PMID- 15804026 TI - [Expression and significance of angiopoietins in oral squamous cell carcinoma]. AB - OBJECTIVE: To observe the expression of angiopoietins in oral squamous cell carcinoma and relationship between the expression of angiopoietins and pathologic classification. METHODS: The expression of Ang-1 and Ang-2 protein in samples from 42 oral squamous cell carcinoma and 16 oral normal mucosa were detected by immunohistochemical technique. RESULTS: The expression of Ang-1 and Ang-2 was found in both oral squamous cell carcinoma and normal control. It was found Ang-1 expression alike in oral squamous cell carcinoma and control (P > 0.05). Ang-2 expressed at low level in control while strongly positive in oral squamous cell caroinoma and the level of Ang-2 expression in oral squamous cell caroinoma was related to pathologic classification (P < 0.05). CONCLUSION: Ang-2 expression in oral squamous cell carcinoma is related to angiogenesis and pathological classification, which is probably involved in angiogenesis regulation, promotes the development and metastasis of oral squamous cell carcinoma. PMID- 15804027 TI - [The effects of HSV-tk suicide gene and wild-type p53 gene on pleomorphic adenoma cells of salivary gland]. AB - OBJECTIVE: To study the therapeutic effects of combined gene therapy of wild type p53 (wt-p53) and herpes simplex virus thymidine kinase (HSV-tk) gene on pleomorphic adenoma cells of salivary gland. METHODS: Wild type p53 and HSV-tk gene were transfected into human pleomorphic adenoma cells of salivary gland by using recombinant adenovirus vector. The efficiency of transfection was checked and gene was expressed by RT-PCR methods. The cell inhibition after transfected was verified by light microscope and MTT. RESULTS: The proliferation of the pleomorphic adenoma cells transfected wt-p53 and HSV-tk gene was inhibited and the cell survival rate decreased to 54% and 38% respectively in 5 days. However, when wt-p53 gene combined with HSV-tk/GCV system, the killing effects was significantly stronger (P < 0.05) and the cell survival rate decreased to 20%. CONCLUSION: Combining p53 gene with HSV-tk/GCV system for gene therapy in pleomorphic adenoma cells of salivary gland is a valuable method. PMID- 15804028 TI - [An experirmental study on the preparation and drug sustained release characteristics of Pingyangmycin Albumin Microspheres]. AB - OBJECTIVE: The aim of this study was to prepare Pingyangmycin Albumin Microspheres (PYM-AMS) for arteriovenous malformations treatment. METHODS: PYM AMS was prepared at 140 degrees C by the method of emulsification-heat solidification and its characteristics were evaluated, such as morphosis, particle size, drug loading (DL%), encapsulation efficiency (EE%), stability and drug sustained-releasing in vitro. After being packaged, PYM-AMS were sterilized with 13.7 kGy of 60Co. Small samples of PYM-AMS were packaged in small bottles and stored at 3 - 5 degrees C, 15 - 25 degrees C, 37 degrees C for 3 months, then checked the change of morphology, DL, EE and the release rate. RESULTS: The surface of particles was smooth and integrated. The average diameter of PYM-AMS particles was 139.422 microm and 80% was in the range of 56 - 251 microm. The mean DL% and EE% were 26.47% and 84.3%, respectively. PYM released fast in 5 h, but then released slowly. 88.65% drugs were released in 24 h, and t50 was 1.5 h. There was no obvious change of the morphology, DL,EE and the release rate of PYM AMS stored at 3 - 5 degrees C 15 - 25 degrees C, 37 degrees C for 3 months. CONCLUSION: PYM-AMS prepared in this study had sustained-release effect, high drug loading and high stability. Albumin is a good carrier of PYM embolization agent. PMID- 15804029 TI - [The effect of dog bilateral distraction osteogensis in temporomandibular joint]. AB - OBJECTIVE: To investigate the effect of bilateral mandibular distraction osteogenesis in the condyles. METHODS: 16 adult hybrid dogs were randomly divided into normal control group and experiment group. Experimental dogs underwent bilateral mandibular osteodistraction at a rate of 1 min/day. 4 dogs were killed respectively in distraction period, 2 and 8 weeks after completion of 10 days distraction. The bilateral condyles specimens were harvested and examined with histological and immunohistochemical methods. RESULTS: Compared with normal control group, various degrees of irregularities and erosion were found in fibrocartilage of condyle in experiment group, including damage in fibrous layer, hyperplasia layer and proliferative layer and osteogenic activity in cartilage layer. A significant increase of TGF-beta1 expression was also found in experiment groups. TGF-beta1 positive staining was noted in hypertrophic cell, matrix and chondroblast, osteoblast and matrix in osteogenic activity areas. These changes were the most obvious in 2 weeks after completion of distraction. CONCLUSION: Gradual bilateral mandibular distraction at a rate of 1 mm/day brought degenerative changes of condyle, but the changes are reversible. PMID- 15804030 TI - [Isolation and identification of huamn dental pulp stem cells]. AB - OBJECTIVE: To isolate and identify human dental pulp stem cells from third molars. METHODS: Dental pulps were dissected and digested by collagenase type I and dispase. The obtained single cell supernatant were harvested and cultured. Characterization of the phenotype of DPSCs was detected by immunohistochemical method and RT-PCR assay. Cell cycle was analyzed by FCM. Differentiation potential of DPSCs was evaluated. RESULTS: Colony-forming efficiency of cells derived from dental pulp tissue was 2 - 15 clones/10(3) cells plated. DPSCs were found to express many different markers, including vimentin, collagen type I, GFAP, nestin and osteocalcin, while they failed to react with MyoD and DSPP. About 64.1% of the cells were in G0/G1 phases, while only 35.8% in proliferation (S + G2 + M). Grown in an adipogenic cocktail medium for three weeks, some DPSCs expressed fat cell markers of PPARgamma and LPL, and formed oil red O-positive lipid clusters in five weeks. After culture with a myogenic-inductive medium, DPSCs were found to express MyoD, desmin and myosin, markers of myocyte. Long term cultures of DPSCs grown in differentiation inductive medium demonstrated the capacity to form Von Kossa-positive condensed nodules with high levels of calcium. CONCLUSION: Cells isolated from adult human dental pulp are clonogenic, and have multipotent differentiation potential, satisfying the criteria of postnatal somatic stem cell. PMID- 15804031 TI - [Biological effects of nano-hydroxyapatite/polyamide 66 on the dental pulp cells]. AB - OBJECTIVE: To investigate the biological effects of the new nano hydroxyapatite/polyamide 66 biological composites (nHA-PA66) on the dental pulp cells. METHODS: After interaction with the nHA-PA66 eluate, the growth, proliferation, and function of the in vitro cultured human dental pulp cells were studied by cell culture technique, inverted phase-contrast microscope observation, MTT assay, flow cytometry, ALP activity assay and quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) analysis. RESULTS: The cultured pulp cells grew well and showed no morphological variation. Moreover, this material had no negative effects on the proliferation, cell cycle, ALP activity and the expression of dentin sialophosphoprotein (DSPP) mRNA of the pulp cells. CONCLUSION: As a new nano-biomaterial, nHA-PA66 has good biocompatibility to the pulp cells, but no obvious bioactivity. PMID- 15804032 TI - [The effect of galla chinensis on the growth of cariogenic bacteria in vitro]. AB - OBJECTIVE: The purples of this study was to investigate the role of different components of Galla Chinensis extract on the growth of 6 kinds of cariogenic bacteria, and to find out the most effective components of Galla Chinensis extract. METHODS: Four different components (GCE1, GCE2, GCE3 and GCE4) were separated from Galla Chinensis and there antibacterial activities to Streptococcus mutans ATCC 25175, Streptococcus sanguis ATCC 10556, Streptococcus salivarius SS 196, Actinomyces naeslundii WVU 627, Actinomyces viscosus ATCC 19246 and Lactobacillus rhamnosus AC 413 were checked. There effects on the growth curve of Streptococcus mutans ATCC 25175 were also investigated. RESULTS: The most effective part of Galla Chinensis was found to be GCE2 and GCE4, which were found to be a mixture of polyphenol-rich fractions. All of the different components had an inhibitory effect to the growth of Streptococcus mutans ATCC 25175. CONCLUSION: All of the 4 different components of Galla Chinensis extract could inhibit the growth of the tested bacteria. These results suggest that the antibacterial activity of Galla Chinensis extract is caused by a synergistic effect of monomeric polyphenols, which can easily bind to proteins. PMID- 15804033 TI - [An article in Lancet set a SBU report rocking. When SBU authors question their own conclusions: Who is to be trusted?]. PMID- 15804034 TI - [Alcohol as medication is no good. More risks than benefits according to a survey of current knowledge]. AB - Moderate drinking has small effects on health. Alcohol-related risks are greatest for young people, and decrease with age. Women are more sensitive than men to the effects of alcohol. Protective effects of moderate drinking are reported for cardiovascular disease, diabetes type 2 and cognitive functioning. However, moderate drinking also involves risks, especially of injuries, violence, foetal damage, certain forms of cancer, liver disease and hypertension. Alcohol consumption should not be recommended for health reasons. Binge drinking, regardless of age, is a medical risk. Health professionals should discuss the pattern of drinking with patients, especially binge drinking, to a larger extent than is usually the case today. PMID- 15804035 TI - [Potential for organ donation in Sweden. Analysis of five-year registration of all ICU deaths in southern Sweden]. AB - A complete registration of all deceased patients at intensive care units in the Southern Health region of Sweden has shown that 3,114 patients died during the five years from 1999 to 2003. Only 174 cases (5.6 per cent) were classified as potential organ donors according to the definition of total brain infarction (brain death) without medical contra-indications against organ donation. Consent for organ donation was given in slightly more than half of these cases. In 42 per cent of the cases relatives were not aware of the attitude of the deceased, and in 40 per cent of these cases they used their right of veto against organ donation. Corresponding registration, as part of the computerised system for quality assurance for intensive care (PASIVA), may become a national and complete quality assurance for organ donation in Sweden. PMID- 15804036 TI - [Carbon monoxide--a toxic gas and ...a signal molecule with therapeutic potential]. AB - The toxic gas, carbon monoxide (CO), is produced endogenously during the catabolism of heme, resulting in the co-production of biliverdin, and iron (Fe2+) in equimolar amounts. This process is catalysed by heme oxygenase (HO). HO exists in three different isoforms. Inducible HO-1 may be induced by a wide variety of stimuli. HO-2 and HO-3 are constitutively expressed. The toxic effects of CO are well known. At low concentrations, CO appears to be cytoprotective. CO shares some chemical and biological properties with nitric oxide (NO), which together with some experimental findings has led to the suggestion that CO also possesses physiological functions; a role as signalling molecule has been proposed. Subsequently, it has been demonstrated that CO is a mediator in the autonomic nervous system and also in the immune system. NO and CO production appears to be reciprocally modulated by the respective synthesizing enzyme, which indicates a complicated interrelationship between the two mediators. PMID- 15804037 TI - [Sexual problems are common during antidepressive treatment. What do general practitioners know about this adverse effect and how is it handled?]. AB - At least five per cent of the Swedish population is treated with some kind of antidepressant (AD) drug, mostly SSRI's. Sexual adverse effects, mainly loss of libido and anorgasmia or delayed orgasm/ejaculation concern more than 50% of these patients. The antidepressive effects of treatment are generally satisfying, but the sexual side effects may be a reason for non-compliance. This article refers to studies that specifically have investigated sexual adverse effects caused by AD-treatment and strategies to handle and prevent these problems. A questionnaire sent to general practitioners showed that they are well aware of the problem with sexual adverse effect in AD treatment and ascribe it great importance. The knowledge of effects and side effects of newer AD drugs seemed somewhat insufficient. An increased knowledge about these substances and alternative treatment strategies would perhaps make it easier for the physicians to handle or avoid AD induced sexual side effects. PMID- 15804038 TI - [Post-exertional respiratory problems in asthma are reduced by exercise]. PMID- 15804039 TI - [Tegner, Strindberg and Froding. Poets under the psychiatric magnifying glass]. PMID- 15804040 TI - [Pfizer's answer to Bengt Jarhult: joint challenge to develop methods for discovery of low-frequency adverse effects]. PMID- 15804041 TI - [Insights and views about cardiovascular safety of COX2 inhibitors]. PMID- 15804042 TI - [Lunchbox, anyone?]. PMID- 15804043 TI - Local delivery of soluble interleukin-6 receptors to improve the outcome of alpha toxin producing Staphylococcus aureus infection in mice. AB - Staphylococcal alpha-toxin enhances interleukin (IL)-6 secretion in mice infected with Staphylococcus aureus. The role of alpha-toxin-induced IL-6 secretion in host defense has not been sufficiently clarified. In the present study, IL-6 signaling was transiently regulated using soluble IL-6 receptors (sIL-6R) to investigate the role of IL-6 in the early stage of abdominal S. aureus infection. In mice challenged with bacteria producing high alpha-toxin levels, the local delivery of sIL-6R was effective in improving the survival rate, the resolution of neutrophilia and the bacteria clearance. Mice that had received sIL-6R and survived showed high levels of IL-6, monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor (TNF)-alpha. In contrast, mice that died in spite of the delivery of sIL-6R showed high levels of interferon (IFN)-gamma and IL-1alpha and low TNF-alpha level. When the effect of soluble gp130, a sIL-6R antagonist, was examined, the number of neutrophils increased significantly and the MCP-1 level decreased significantly, compared to the group that received sIL-6R alone; the number of viable bacteria also tended to increase as a result of the inhibition of IL-6 signaling. The cellular phosphotyrosine level in alpha-toxin-treated macrophages was reduced in cultures supplemented with recombinant IL-6 in vitro. These results suggest that IL-6 enhances bactericidal activity and reduces the number of immune cells that are activated abnormally through the regulation of inflammatory cytokines during the early stage of infection in alpha-toxin producers. PMID- 15804044 TI - Impairment of the splenic immune system in P2X(2)/P2X(3) knockout mice. AB - The isolated spleens from male and female mice lacking P2X(2) and P2X(3) receptors (P2X(2)/P2X(3) knockout (KO) mice) and those from wild-type (WT) mice were investigated by flow cytometry, immunohistochemistry and functionally by organ-bath pharmacology. The spleens from the P2X(2)/P2X(3) KO mice weighed significantly more than the corresponding WT mice. Flow cytometry was used to isolate the mononuclear cells, which were then phenotyped. T-lymphocytes, B lymphocytes and macrophages were identified and counted. It was found that the increase in size of the spleens from the KO animals corresponded to an increase in the numbers of mononuclear cells present and that all three cell types (T lymphocytes, B-lymphocytes and macrophages) increased in much the same proportion as those from the WT animals. Immunohistochemical localisation of P2Y(1), P2Y(2) and P2X(1) receptors revealed their presence on the spleen capsule and trabeculae. P2X(1) receptors were also present on blood vessels. There was no difference in the expression of these receptors between the WT and P2X(2)/P2X(3) KO spleens. Functional studies revealed the presence of multiple P2 receptors inducing the contraction of the spleen capsule, from both WT and KO mice. There was no difference in the contractions induced by adenosine 5'-triphosphate (ATP), alpha,beta-methylene ATP, 2-methylthio ADP or uridine triphosphate from WT and KO mice. It is concluded that mice lacking both P2X(2) and P2X(3) receptors have enlarged spleens and that this is correlated with an increase in the number of immune cells, perhaps as a consequence of a compromised immune system and chronic infection. PMID- 15804045 TI - Blockade of TNF does not alter oxygen burst and phagocytosis of human neutrophils in patients with rheumatoid arthritis. AB - Clinical trials evaluating tumor necrosis factor alpha (TNF-alpha) binding agents in patients with rheumatoid arthritis (RA) have demonstrated significant efficacy in reducing symptoms of disease and slowing radiographic progression. However, infectious complications are the most severe and common adverse effects of anti TNF therapy. The functional capacities of neutrophils (PMNs) as the first line of defense in bacterial and fungal infections are enhanced by soluble TNF as a potent neutrophil primer. The aim of this study was to assess the influence of in vivo TNF blockade on oxygen burst (OB) and phagocytosis of human neutrophils. PMNs were derived from 20 patients with RA on anti-TNF-alpha therapy and 13 patients using conventional DMARDs. By flow cytometry we measured OB upon stimulation with Escherichia coli and N-formyl-1-methionyl-1-leucyl-phenylalanine (FMLP) with and without priming with granulocyte-colony stimulating factor (G CSF) and/or TNF-alpha using dihydrorhodamine (DHR) 123. Phagocytosis of fluorescein isothiocyanate (FITC)-labeled E. coli was also assessed by flow cytometry. Thirty-three healthy volunteers served as controls. Upon stimulation with E. coli and FMLP, there was no significant difference in OB between the two patient groups and healthy controls. Priming was effective in all groups. Phagocytosis of E. coli by PMNs was equally effective in controls and patients independent from the treatment regimen. These data show that OB, phagocytosis and responsiveness to priming with TNF and G-CSF of PMNs are not impaired in patients with RA treated with anti-TNF agents in comparison with patients on conventional DMARDs or healthy controls. Thus, the infectious complications observed in patients with TNF blockade cannot be explained by functional impairment of PMNs; however, the neutralization of TNF as a potent primer of neutrophil response may increase the susceptibility for infections in these patients. PMID- 15804046 TI - The effect of intestinal colonization of germ-free pigs with Escherichia coli on calprotectin levels in plasma, intestinal and bronchoalveolar lavages. AB - Calprotectin levels were measured by ELISA in plasma, terminal small bowel lavage and bronchoalveolar lavage from 8-day-old germ-free piglets or gnotobiotic piglets 24 h after colonization with one of the following Escherichia coli strains: non-pathogenic O86, probiotic Nissle 1917 or enteropathogenic O55. The concentration of calprotectin in plasma was about 30 ng/ml only in germ-free piglets and piglets associated with non-pathogenic E. coli. Piglets infected with O55 showed a significant increase of plasma calprotectin and the highest mean level of calprotectin in the bronchoalveolar lavage, which was coincident with septicaemia. However, in the lumen of the small intestine, E. coli Nissle 1917 alone elicited a significant increase of the calprotectin level which was confirmed by immunofluorescence and APAAP immunohistochemistry on cryostat sections through the small bowel. The relevance of this finding to the therapeutic effect of E. coli Nissle 1917 in inflammatory bowel disease is discussed. PMID- 15804048 TI - Recent advances in hereditary spinocerebellar ataxias. AB - In recent years, molecular genetic research has unraveled a major part of the genetic background of autosomal dominant and recessive spinocerebellar ataxias. These advances have also allowed insight in (some of) the pathophysiologic pathways assumed to be involved in these diseases. For the clinician, the expanding number of genes and genetic loci in these diseases and the enormous clinical heterogeneity of specific ataxia subtypes complicate management of ataxia patients. In this review, the clinical and neuropathologic features of the recently identified spinocerebellar ataxias are described, and the various molecular mechanisms that have been demonstrated to be involved in these disorders are discussed. PMID- 15804049 TI - Diagnostic immunohistology of muscle diseases. AB - The diagnostic muscle biopsy has seen the use of virtually every histologic technique in existence over the past 50 years. Since the 1960s, enzyme histochemistry has become the chief technique in evaluating muscle biopsies. However, the increasing knowledge of cellular constituents and associated connective tissue of the myofiber coupled with the increasing availability of a broad diversity of antibodies to these proteins promises to bring the diagnosis of muscle disease to the same state of dependency upon immunohistochemistry as in the contemporary pathologic diagnosis of neoplasia. Immunohistochemistry may be used for both the identification of normal antigenic constituents in skeletal muscle and their loss, accumulation, or maldistribution in corresponding myopathies, sometimes with small biopsies or lacking frozen tissue, in paraffin sections. Three broad categories of muscle diseases will be characterized in terms of diagnostic antibodies in current use: dystrophic, congenital/structural, and inflammatory myopathies. PMID- 15804047 TI - Specific binding of L-ficolin and H-ficolin to apoptotic cells leads to complement activation. AB - The serum lectins mannose-binding lectin (MBL), L-ficolin, and H-ficolin are recognition molecules in the lectin complement pathway, which play an important role in innate immunity. To assess involvement of the lectin pathway in the clearance of apoptotic cells, we used flow cytometry to quantify binding of MBL, L-ficolin, and H-ficolin to apoptotic HL60, U937, and Jurkat cells induced by actinomycin D. When apoptotic cells were incubated with normal human serum, MBL and L-ficolin bound to all three cell lines tested; moreover, H-ficolin bound to apoptotic Jurkat cells only. Subsequently, C4 and C3 were deposited on apoptotic cells of all three cell lines. MBL, L-ficolin, and H-ficolin binding to apoptotic cells was confirmed by the use of purified proteins. Purified C4 added to apoptotic cells that had bound pure L-ficolin was deposited on the cell surfaces. In L-ficolin-depleted serum, C3 deposition on HL60 or Jurkat cells decreased to approximately 50% or 70%, respectively, in comparison to the serum before L ficolin depletion. We conclude that L-ficolin, in addition to MBL, recognizes apoptotic cells and activates complement via the lectin pathway. We also observed in vitro binding of L-ficolin and H-ficolin to cC1q receptor (C1q receptor specific for the collagenous region of C1q)/calreticulin, a candidate receptor for the collagenous region of MBL and C1q. Thus, L-ficolin and H-ficolin as well as MBL participate in the clearance of apoptotic cells through complement activation. PMID- 15804050 TI - Cell proliferation and granule cell dispersion in human hippocampal sclerosis. AB - Granule cell dispersion (GCD) is observed in approximately 40% of cases of hippocampal sclerosis (HS) in patients with epilepsy. Studies in animal models suggest that GCD may be a consequence of enhanced proliferation of granule cell precursors as a result of seizures. We quantified the number of cells in cycle in subfields of the hippocampus with immunohistochemistry for Mcm2 in 14 HS cases with or without severe GCD compared to 6 epilepsy patients without classical HS or GCD as well as 5 postmortem controls. Higher numbers of Mcm2-positive cells were seen in the region of the granule cell layer in patients with severe GCD, and immunolabeling with Geminin and Ki-67 confirmed a proportion were progressing through cycle. Double labeling with Mcm2 and GFAP confirmed the majority of these cycling cells were GFAP-negative and occasional cells stained colocalized with stem cell marker nestin. These findings support the view that GCD may be a phenomenon related to increased progenitor cell proliferation in patients with hippocampal damage and chronic epilepsy. PMID- 15804051 TI - 14-3-3 protein in CSF: an early predictor of SIV CNS disease. AB - In neurons, 14-3-3 proteins regulate diverse processes, including signal transduction, neurotransmitter production, and apoptosis by binding to target proteins, but the role 14-3-3 proteins play in the pathogenesis of central nervous system (CNS) disease remains unclear. To examine the relationship between presence of 14-3-3 protein in cerebrospinal fluid (CSF) and encephalitis in the SIV/macaque model of HIV CNS disease, CSF levels of 14-3-3 protein were measured by quantitative immunoblotting throughout infection in 6 SIV-infected pigtailed macaques. Beginning during asymptomatic infection and continuing until death, CSF levels of 14-3-3 were elevated in 4 of 6 SIV-infected animals. Animals with 14-3 3 protein in CSF had the highest viral loads in the CSF after acute infection and the highest levels of both viral RNA and protein in brain (p < 0.001). In contrast, the presence of 14-3-3 protein in CSF was not associated with CNS microglial/macrophage activation measured by quantitative immunohistochemical staining for CD68 (p = 0.13). CSF levels of 14-3-3 protein may be a valuable marker of early neuronal damage, CNS viral replication, and CNS disease progression in HIV-infected individuals. PMID- 15804052 TI - Increased frequency of argyrophilic grain disease in Alzheimer disease with 4R tau-specific immunohistochemistry. AB - Argyrophilic grain disease (AGD) is a medial temporal 4R tauopathy with filamentous inclusions in dendrospinal portions of neurons. AGD is often associated with mild Alzheimer-type pathology, but it is difficult to detect AGD in the setting of advanced Alzheimer disease (AD). The frequency of AGD in AD has been difficult to determine because of masking of grains by neurofibrillary lesions. To address this issue, medial temporal lobe sections from AD brains were immunostained with a 4R tau-specific antibody, ET3, which permitted detection of grains even in the setting of advanced neurofibrillary degeneration. AGD was found in 61 of 239 AD cases (26%). The frequency of AGD in AD in this study is higher than in previous studies that relied on less selective staining methods, such as the Gallyas silver stain or immunostaining with phospho-tau antibodies. The frequency of AGD in AD did not correlate with Braak stage or with the density of neurofibrillary tangles and senile plaques in the limbic lobe; however, AD cases with AGD were significantly older than cases without AGD. The MAPT H1 frequency tended to be higher in AD cases with AGD than in those without AGD, but there were no differences in APOE epsilon4 carrier state. These findings suggest advanced age and possibly MAPT H1 are risk factors for AGD, even in the setting of concurrent AD, in which neurofibrillary degeneration is associated with accumulation of both 3R and 4R tau. PMID- 15804053 TI - Implication of fetal SMN2 expression in type I SMA pathogenesis: protection or pathological gain of function? AB - Spinal muscular atrophy (SMA) is caused by mutations in the survival motor neuron gene 1 (SMN1). The SMN2 gene, which is the highly homologous SMN1 copy that is present in all the patients, is unable to prevent the disease. Most of the SMN1 transcript is full-length, whereas a substantial proportion of the SMN2 transcript lacks exon 7 (delta7). We characterized the developmental expression of SMN2 by comparing control and SMA fetuses. The control spinal cord revealed the highest amount of FL SMN, most of which was of SMN1 origin. When analyzing the SMA spinal cord transcripts, we detected a considerable reduction in the FL/delta7 ratios due to a decrease in the FL and an increase in delta7 isoform. After immunoblot and immunohistochemistry analyses, we found that the amount of SMN2 protein in the SMA spinal cord and muscle was lower than in the controls. However, the results of the expression of SMN2 in intestine, lung, adrenal gland, kidney, and eye, which are unaffected by the disease, were the same in controls and SMA samples. In these tissues, SMN2 may compensate for the absence of SMN1, whereas in SMA motor neurons, a cell-specific dysregulation of the SMN2 expression could favor the onset of the acute form of the disease. PMID- 15804054 TI - Selective neuron loss in the paraventricular nucleus of hypothalamus in patients suffering from major depression and bipolar disorder. AB - The structural basis for depressive disorders remains unknown. Studies using neuroimaging and postmortem brain tissue indicate that anatomic substrates may contribute to major depression disorder (MDD) and bipolar disorder (BD). The present study used design-based stereology to assess neuron loss in 2 well defined hypothalamic structures, the paraventricular nucleus (PVN) and supraoptic nucleus (SON), in clinically well-studied cases with severe depression. The left or right diencephalon was blocked from 26 brains removed at autopsy from age matched controls (5 male/3 female) and patients with MDD (6 male/5 female) and BD (5 male/2 female). Serial sections were cut at an instrument setting of 60 microm through the entire PVN and SON from left hypothalamus and 8 to 10 sections per brain were systematic-random sampled and stained with cresyl violet. A trained operator blind to clinical diagnosis used computerized stereology to estimate total neuron numbers in PVN and SON. The results revealed a selective, robust reduction of approximately 50% in total neuron number in the PVN for major depression and bipolar cases compared with age-matched controls, with no differences in neuron numbers in the SON. This selective neuronal loss in the PVN appears to identify an important neurobiologic substrate for the behavioral manifestations of depression. PMID- 15804055 TI - Corticospinal regeneration into lumbar grey matter correlates with locomotor recovery after complete spinal cord transection and repair with peripheral nerve grafts, fibroblast growth factor 1, fibrin glue, and spinal fusion. AB - Knowledge of which tracts are essential for the recovery of locomotor function in rats after repair is unknown. To assess the mechanism of recovery, we examined the correlation between functional recovery and axonal regeneration. All rats underwent complete cord transection and repair with peripheral nerves, fibroblast growth factor 1, fibrin glue, and spinal fixation. Repaired rats recovered both motor-evoked potentials recorded at the lumbar level and locomotor function. Cord retransection rostral to the repair abolished the recovery, indicating improvement was due to long tract regeneration. To determine which long tracts correlated with recovery, a novel technique of simultaneous bidirectional axonal tracing and immunohistochemical examination of axonal type was used to quantitate the regeneration of corticospinal, rubrospinal, reticulospinal, vestibulospinal, raphespinal, propriospinal, serotonergic, and calcitonin gene-related peptide containing axons. Multiple linear regression analysis revealed recovery of function correlated only with regeneration of corticospinal axons into the gray matter of the lumbar spinal cord (R = 0.977, p < 0.02). For the first time, we show that regeneration of the corticospinal tract into the lumbar gray matter is a mechanism of functional locomotor recovery after complete cord transection and repair. PMID- 15804056 TI - Tau and alpha-synuclein inclusions in a case of familial frontotemporal dementia and progressive aphasia. AB - Recent studies have shown that neurofibrillary tangles are frequently accompanied by alpha-synuclein inclusions in sporadic and familial Alzheimer disease, in Down syndrome, in progressive supranuclear palsy, and Parkinsonism dementia complex of Guam. Here we report the cases of 2 brothers with familial progressive aphasia who developed features of frontotemporal dementia with predominant tau pathology but also alpha-synuclein pathology. The 2 patients' brains revealed abundant tau pathology in the hippocampus and basal ganglia, whereas tau and alpha-synuclein aggregates coexisted only in the nucleus basalis of Meynert, the only region where alpha-synuclein was present. In this brain region, abundant Lewy bodies, Lewy neurites, and tau inclusions were found; the pathology was more abundant in the older than in the younger brother. Sarkosyl-insoluble tau extracted from brains of the 2 patients showed the presence of tau filaments that contained 3 major tau bands of 60, 64, and 68 kDa on Western blot analysis. These bands contained mainly tau with 3 and 4 repeats and no amino-terminal inserts and tau with 4 repeats and one amino-terminal insert. No mutations were identified in the tau, alpha-synuclein, beta-synuclein, or parkin genes. We think that this is the first report showing a specific colocalization of neurofibrillary tangles and Lewy bodies in a family with progressive aphasia. PMID- 15804057 TI - Inflammatory myofibroblastic tumor of the central nervous system: clinicopathologic analysis of 10 cases. AB - To verify the pathologic features, anaplastic lymphoma kinase (ALK) expression and biologic behavior of inflammatory myofibroblastic tumors (IMTs) of the central nervous system (CNS), we analyzed 10 cases of IMTs-CNS (8 cranial, 1 spinal, and 1 orbital). Our series of IMTs of the CNS showed a male predominance (male:female = 6:4) and a wide age range (10-60 years; mean age, 46.7 years). Lesion location also varied, but they were basically dura-based. Radiologically, they showed two patterns: isolated mass forming (n = 6) and an en plaque-like pattern (n = 4). Histopathologically, plasma cell granuloma (PCG)-like (n = 5) or fibrohistiocytic (FHC) variant (n = 5) was present. No correlation was found between the radiologic and histopathologic patterns. Spindle-shaped mesenchymal cells of IMTs expressed smooth muscle actin (SMA) in all cases. ALK expression was not found in our IMTs of the CNS. Late recurrence was found in 2 cases in different sites (20%). Pathologically, IMT-CNS could be subclassified into PCG like and FHC. Immunostaining for SMA was found to helpfully discriminate myofibroblastic cells and to make a differential diagnosis. Although our cases did not show ALK immunoreactivity, some IMTs-CNS can recur, which suggests the neoplastic potential of these tumors. The rearrangement of the ALK gene in IMTs CNS should be verified by an examination of more cases. PMID- 15804058 TI - Diagnostic value of carboxypeptidase-H autoantibodies in detecting latent autoimmune diabetes in adults. AB - OBJECTIVE: To investigate the diagnostic value of carboxypeptidase-H (CPH) autoantibodies in Chinese patients with the latent autoimmune diabetes in adults (LADA). METHODS: One hundred and fifty-four Type 1 diabetes, 104 Type 2 diabetes, and 144 healthy people were enrolled. Recombinant human CPH (54 kD) was labeled by in vitro translation with 35S-methionine and used to evaluate autoantibodies to CPH (CPH-Ab). Radioimmunoassay was applied to detect antibodies to glutamic acid decarboxylase (GAD-Ab), intracellular part of protein tyrosine phosphatase like protein (IA2ic-Ab), and autoantibodies to insulin (IAA). RESULTS: No differences in CPH-Ab prevalence were found among Type 1 diabetic patients (5/154, 3.2%), Type 2 diabetic subjects (6/104, 5.7%),and the healthy controls (3/144, 2.1%). The prevalences of GAD-Ab, IA2ic-Ab, and IAA were 15.4% (16/104), 2.9% (3/104), and 2.3% (1/43), respectively in Type 2 diabetes. All IA2ic-Ab or IAA-positive patients with Type 2 diabetes were GAD-Ab-positive. No GAD-Ab- or IAA-positive subjects were observed in CPH-Ab-positive patients with Type 2 diabetes. CONCLUSION: CPH-Ab may provide some diagnostic value for LADA, and improve the sensitivity in diagnosing LADA in Chinese when combined with GAD-Ab test in Type 2 diabetes. PMID- 15804059 TI - [Angiotensin converting enzyme gene polymorphism and type 2 diabetic nephropathy]. AB - OBJECTIVE: To prospectively clarify the relationship between angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and diabetic nephropathy (DN) in Type 2 diabetic patients. METHODS: We examined 66 Type 2 diabetic patients with normal buminure. All patients suffered from diabetes mellitus for more than 5 years and matched well in age, body mass index (BMI), mean arterial pressure (MAP), fasting blood sugar (FBS), and urinary excretion of albumin (UAE). Patients were classified into 3 groups according to genotypes of ACE. ACE gene I/D polymorphism was identified by polymerase chain reaction (PCR). The patients were followed up for 5 years and their UAE, serum creatinine (Scr), and HbAlc were checked once a year. RESULTS: Eight subjects (12%) were discontinued prematurely. At the end of the study, there were no significant differences in the clinical parameters such as BMI, MAP, FBS, HbA1c, UAE, and Scr among the 3 groups (P > 0.05), and also in DN incidence in Type 2 diabetic patients among the 3 groups (II 45.8%, ID 52.3%, and DD 46.1%, respectively, P > 0.05 ). The increased degree of UAE among the 3 groups was similar at the end point of the study (P > 0.05). CONCLUSION: The DD genotype of ACE gene may not be a clinically useful genetic marker for predicting the genesis and development of DN in Type 2 diabetic patients in Chinese. There is no association between ACE gene I/D polymorphism and the genesis and development of DN in Type 2 diabetic patients. PMID- 15804060 TI - [Indomethacin-induced HL-60 leukemic cell apoptosis and the activation of C-jun NH2- terminal kinase signal transduction pathway]. AB - OBJECTIVE: To observe the effect of indomethacin on inhibiting the proliferation of HL-60 cells and inducing HL-60 cells apoptosis and to explore the activation of C-Jun NH2-terminal kinase (JNK) signal transduction pathway in mediating indomethacin-induced HL-60 cell apoptosis. METHODS: The cell viability was determined by Trypan blue staining and the state of cell proliferation was analyzed; DNA ladder pattern and AO/EB staining were applied to identify the cell apoptosis; the apoptotic signal proteins including caspase-9, caspase-3, and PARP and the proteins of JNK signal pathway such as MEK4, JNK, P-JNK, and P-C-Jun, were detected by Western blotting. RESULTS: Indomethacin at 200 to approximately 400 micromol significantly inhibited the proliferation of HL-60 cells and induced the cell apoptosis in a time- or concentration-dependent manner; caspase-9, caspase-3, and PARP were cleaved and activated in undergoing apoptotic cells; and the expressions of MEK4, P-JNK, and P-C-Jun were upregulated with the increase of indomethacin concentration. CONCLUSION: Indomethacin can inhibit the proliferation of HL-60 cell and induce leukemic cell apoptosis. The activation of JNK signal transduction pathway mediates the event of indomethacin-induced HL-60 cell apoptosis. JNK signal pathway is located in the upstream of caspase signal pathway. PMID- 15804061 TI - [Expression and transcription of inducible nitric oxide synthase in the liver ischemic preconditioning in rats]. AB - OBJECTIVE: To investigate the changes in transcription and expression of inducible nitric oxide synthase (NOS2) in the liver ischemic preconditioning (IP), and to determine the role of nitric oxide (NO) synthetic pathway in the liver IP in rats. METHODS: We randomly divided 131 Sprague Dawley rats into 3 groups: ischemia/reperfusion (I/R) group (n=52), IP group (n=41), and sham operation (S) group (n=38). Plasm NO concentration and the transcription and expression of NOS2 were detected 2 hours, 24 hours, and 1 week after the operation. RESULTS: (1) In the IP group, the NO concentrations at the 2nd hour, the 24th hour, and 1 week were significantly higher than those in the S group (P < 0.05, P < 0.01, P < 0.01, respectively) and the NO concentrations at the 2nd hour and the 24th hour were obviously higher than those in I/R group (all P < 0.01). In the I/R group, the NO concentration at the 2nd hour was significantly lower than that in the S group (P < 0.05); there was no significant difference between the I/R group and the S group 24 hours after the operation (P > 0.05), and the NO concentration 1 week after the operation was obviously higher than that in the S group (P < 0.05). (2) In the IP group, the transcription of NOS2 2 and 24 hours after the operation were significantly increased compared with that in the I/R group (all P < 0.05) , but after 1 week, the transcription was not statistically different between the IP group and the I/R group (P > 0.05). (3) In the IP group, the expressions of NOS2 after 2 and 24 hours were obviously higher than those in the I/R group (P < 0.05) and the S group (P < 0.05), but the expression between the IR group and the S group was not significantly different (P > 0.05); after 1 week, the expressions of NOS2 in IP group and I/R group were weakly positive (P > 0.05) , and those in S group were negative (P > 0.05). Conclusion The transcription and the expression of liver NOS2 increase after the liver received ischemic preconditioning in rats. That the peak phases of transcription and expression of NOS2 are ahead of time may be related to the early protective effect of the liver IP. PMID- 15804062 TI - [Heat shock pretreatment inhibits the release of Smac from mitochondria and decreases H2O2-induced cardiomyocyte apoptosis]. AB - OBJECTIVE: To explore the effect of heat shock pretreatment on H2O2-induced apoptosis of neonatal rat cardiomyocytes and the release of Smac from mitochondria. METHODS: After heat shock pretreatment (42 degrees C for 1 h), neonatal cardiomyocytes were exposed to H2O2 (0.5 mmol/L) for 6, 12, 24, and 36 h, respectively. The apoptotic morphological changes and percentage of apoptotic nuclei were analyzed. Activities of caspase-3, 9 were assayed with caspase colorimetric assay kit and Western-blotting. Inducible heat shock proteins were detected by Western-blotting analysis. The release of Smac from mitochondria to cytoplasm was observed by Western-blotting and immunofluorescence. RESULTS: (1) H2O2 (0.5 mmol/L) resulted in a marked release of Smac from mitochondria to cytoplasm at 2 h after the treatment, the activation of caspase-9 and caspase-3 at 4 h after the treatment and specific morphological changes of apoptosis at 24 h after the treatment. (2) Heat shock pretreatment (42 degrees C, 1 h) could increase the expression of hsp90, hsp70 and betaB-crystallin, and inhibit the H2O2-induced release of Smac from mitochondria, the activity of caspase-9, caspase-3 and apoptosis of cardiomyocytes. CONCLUSION: (1) Mitochondrial signal transduction pathway is involved in the apoptosis of cardiomyocytes induced by H2O2; (2) Heat shock pretreatment can inhibit the release of Smac from mitochondria and the activities of caspase-9, 3 and protect cardiomyocytes against H2O2-induced apoptosis. PMID- 15804063 TI - [Protective effects of phosphoramidon on the experimental myocardial injury in rats]. AB - OBJECTIVE: To investigate the protective effects of phosphoramidon on the experimental myocardial injury in rats. METHODS: Thirty-six Wistar rats were randomly divided into 3 groups of 12: the control group (Control), the isoproterenol group (ISO) and the protection group (PA). All rats were anesthetized after 24 hours and the blood was sampled from the carotid artery. The activities of creatine phosphokinase (CPK) and lactic dehydrogenase (LDH) in the serum and the contents of ET in the plasma were detected; then the hearts were isolated and the total layers of apical myocardium were solidified, embedded, sectioned, stained with HE and graded pathologically. RESULTS: The serum activities of CPK and LDH, and the plasmatic content of ET in the ISO group were significantly higher than those in the control group and PA group (P < 0.05); the degree of myocardial pathological damage in the ISO group was significantly higher than that in the control group (P < 0.01), while the degree of damage in the PA group was lower than that in the ISO group (P < 0.05). CONCLUSION: Endothelin is involved in the pathologic process of experimental myocardial injury in rats. Phosphoramidon can protect the myocardial injury in rats and its mechanism may be to inhibit ECE and reduce the endothelins production. PMID- 15804064 TI - [Construction of high efficient eukaryotic expression recombinant on human hepatocyte DNA synthetic stimulated factor]. AB - OBJECTIVE: To construct the eukaryotic expression recombining vector on human The pGEM-hepatocyte DNA synthetic stimulated factor (hHDSSF) with gene cloning. METHODS: hHDSSF, a mid-recombining vector, was constructed by T-A cloning. After restriction endonuclease Not I digestion, the target fragment was subcloned into eukaryotic vector pcDNA3. 1hisB to construct the eukaryotic expression recombinant pcDNA3. 1hisB-HDSSF. RESULTS: The forward insert recombinant pcDNA3. 1hisB-HDSSF was screened and obtained with restriction endonuclease Kpn I digestion and it was detected by DNA sequence analysis. CONCLUSION: The eukaryotic expression recombinant pcDNA3. 1hisB-hHDSSF on hHDSSF is constructed successfully, which lays a foundation for building a stable eukaryotic expression cell strain and expressing hHDSSF. PMID- 15804065 TI - [Construction of recombinant eukaryotic expression vector of NKG2D gene and its expression in COS-7 cells]. AB - OBJECTIVE: To construct recombinant eukaryotic expression vector of NKG2D, and to examine its expression in COS-7 cells. METHODS: Human NKG2D cDNA was obtained from peripheral blood mononuclear cells using RT-PCR, and then the target gene was cloned into pMD18-T vector. A eukaryotic expression plasmid containing target gene was constructed by DNA recombinant technique, and was confirmed by double restriction enzymes digestion and DNA sequence analysis. The recombinant plasmid with encapsuled lipofectamine was transfected into COS-7 cells, and the transfected COS- The 7 cells containing expressive target gene were confirmed by RT-PCR and flow cytometry. RESULTS: The sequence of NKG2D cDNA obtained from the recombinant eukaryotic expression vector was identical with that published on GeneBank. The NKG2D gene was expressed successfully in COS-7 cells. CONCLUSION: The recombinant expression plasmid containing NKG2D gene is constructed successfully. After being transfected to COS-7 cells, the NKG2D protein is expressed by the engineering COS-7 cells line, which lays a foundation for further studying biological activity of NKG2D. PMID- 15804066 TI - [Expression and purification of human metallothionein-1E gene in Escherichia coli]. AB - OBJECTIVE: To express and purify human metallothionein-1E (MT-1E) fusion protein in vitro. METHODS: The cDNA encoding human MT-1E was amplified by RT-PCR and cloned into prokaryotic expressing vector pQE40. After transforming it into Escherichia Coli M15, we determined the solubility of target protein by western blotting and investigated the IPTG inducing condition. The target protein was purified from lysates with Ni-NTA agarose column. RESULTS: Western blotting analysis suggested that both the soluble and insoluble fusion protein existed in Escherichia Coli, but the insoluble was the main expression form. Induced by 1 mM IPTG, the expression of target protein increased with the prolongation of induction time. In our study, after being induced for 8h, the target protein accounted for about 32% of the total bacterial protein. Purified protein was obtained by affinity chromatography. CONCLUSION: We have obtained purified human MT-1E fusion protein, which lays the foundation for the antibody preparation and further functional study. PMID- 15804067 TI - [Establishment and characteristics of the human diploid cell strain HLF-02]. AB - OBJECTIVE: To establish the HLF-02 and provide a new material to the study of geronics, oncology, virology, pharmacology, and biogenetics. METHODS: The lung tissue was digested with 0.15% trypsin. More than 90% of the dissociated cell exhibited initial viability (trypan blue dye exclusion and the ability of adhesion). The surviving cells were analyzed by morphology, growth curves, karyotype analysis, electron microscopic observation and heterotransplantation. RESULTS: A newly established cell strain, which was designated HLF-02 had been maintained in continuous culture for over 50 generations for 10 months. Morphological observation, electron microscopic observation, karyotype analysis and heterotransplantation showed that it was corresponding to diploid cells. The doubling time of the cells was 75.5 hours. Chromosome analyses showed the human cell strain to be typical diploid appearance, and the tumorigenecity assays of nude mice were negative. CONCLUSION: The HLF-02 is a human dipolid cell strain and can serve the further studies of cell gerontics, oncology, virology, pharmacology, and biogenetics. PMID- 15804068 TI - [Evaluation of bone structure and quality of ovariectomized rats by microcrack]. AB - OBJECTIVE: To compare microcrack with bone mineral desity (BMD), bone histomorphometry and biomechanics parameters, and to investigate the potential of microcrack in the evaluation of bone biomechanical quality. METHODS: Eight 10 month-old Sprague-Dawley rats were served as baseline controls, and 90 10-month old rats were randomly divided into A, B, and C groups. Each group comprised ovariectomized (OVX), 17 beta-estradiol treated [EST, 10 micro/(kg x d)] and sham operated (SHAM) subgroups. Rats from groups A,B and C were killed at the 3rd, 15th and 21st week post-operatively. Total body and lumbar vertebral BMD were measured before being killed, and BMD of isolated lumbar vertebrae and tibiae were measured after killing. Bone histomorphometry of the proximal end of isolated right tibia was performed,and compression test was carried out on the isolated 5th lumbar vertebra (L5). After fatigue damage, the isolated 4th lumbar vertebra was stained by en bloc basic fuchsin staining, and microcrack density (Cr. Dn) and microcrack surface density (Cr. SDn) were de- termined on the bone tissue sections. Bone parameters in each subgroup of rats were observed at different time. RESULTS: (1) At the 15th and 21st week post-operatively, multi part BMD, Cr. Dn and Cr. SDn were higher than those at the 3rd week. (2) At the 15th week, trabecular separation (Tb. Sp) increased, trabecular number (Tb. N) decreased, and the maximum loading level and elastic modulus of vertebra reached the peak. (3) At the 3rd week, Tb. Sp, Cr. Dn and Cr. SDn in the OVX subgroup were greater than those in the EST subgroup, while the percentage of trabecular area (TbTr) in the OVX subgroup was lower than that of the EST and SHAM subgroups. No changes of BMDs and biomechanic parameters were observed among the three subgroups. (4) At the 15th week, multi-part BMD and maximum loading level in the OVX and EST subgroups were lower than those in the SHAM subgroup, while elastic modulus, bone histomorphometry parameters, Cr. Dn and Cr. SDn had no change among the three subgroups. (5) At the 21st week, multi-part BMDs, Tb. N and TbTr in the OVX subgroup were smaller than those in the EST and SHAM subgroups. Tb. Sp, bone formation rate, mineral apposition rate, percent labeled perimeter,Cr. Dn and Cr. SDn in the OVX subgroups were greater than those in the EST and SHAM subgroups. Maximum loading level and elastic modulus of vertebra in EST and OVX subgroups were lower than those in the SHAM subgroup. There were no significant differences in all of these parameters Microcrack can be regarded as an alterative between the EST and the SHAM subgroup. Conclusion parameter in the evaluation of bone biomechanical quality. PMID- 15804069 TI - [Quality control of radioligand assay for glutamic acid decarboxylase antibody]. AB - OBJECTIVE: To supply the quality control for glutamic acid decarboxylase antibody (GAD-Ab) measurement using radioligand assay. METHODS: The quality control of GAD Ab determined with radioligand assay in 83 batches was analyzed. RESULTS: The intra-assay coefficient of variation (CV) was 4.3% to approximately 8.3% and the inter-assay CV was 7.1% to approximately 10.8%. A ratio of signal to noise (S/N) with 15 or more was defined as a valid measurement. Of the 83 batches, 67 batches (80.7%) met with the standard and the other 16 (19.3%) were not qualified. When the results were analyzed according to the season, the failure rate in summer was significantly higher than that in winter (38.5% vs 4.8%, P < 0.05). In addition, the mean value of S/N in summer was significantly lower than that in winter ( 14.1 +/- 7. 3 vs. 21.5 +/- 5.2, P < 0.01). Among the failed batches, 68.8% (11/16) was caused by TBST buffer and 50% (8/16) due to the increased temperature of TBST buffer. Other causes for the failure included the invalidation of reagents for in vitro transcription and translation. CONCLUSION: TBST buffer and other influential factors of radioligand assay in determining GAD-Ab, should be controlled strictly to ensure a reliable and repeatable result. PMID- 15804070 TI - [Technique for undecalcified bone tissue. Immunohistochemistry and in situ hybridization]. AB - OBJECTIVE: To investigate the feasibility of applying immunohistochemistry (IH) and in situ hybridization (ISH) technique to the undecalcified bone sections. METHODS: The proximal tibiae of SD rats were embedded with the modified methylmethacrylate low-temperature embedding method and then made into undecalcified bone sections. The sections were stained with Goldner's Masson Trichrome, and the expression of IGF-1 mRNA and protein in the bone tissue cells were detected by IH and ISH. RESULTS: Goldner's staining demonstrated the structure of trabeculae bone was intact and osteoid at the edge of trabeculae stained red. Osteoblasts and osteoclasts in bone sections could be seen clearly. The positive signals of IGF-1 mRNA and protein were found in the cytoplasm of osteoblasts, chondrocytes in epiphyseal plate and some mononuclear cells in the bone marrow. The matrix of trabeculae also showed positive expression of IGF-1 protein. CONCLUSION: The undecalcified bone sections prepared by the method can meet the needs of IH and ISH. The establishment of this method will provide technological platform for the study of molecular pathology on metabolic bone diseases such as osteoporosis. PMID- 15804071 TI - [Serum leptin and insulin resistance in obesity and effects of sibutramine on them]. AB - OBJECTIVE: To assess the relationship between serum leptin and insulin resistance (IR) in obesity, and to investigate the effects of sibutramine on obesity, serum leptin and IR. METHODS: Seventy obese subjects [body mass index (BMI) > or =25 kg/m2] were randomly divided into 2 groups: group B (sibutramine 10 mg/day) and group C (a placebo tablet/day). Both had been treated for 12 weeks. Another 30 healthy adults served as the normal control (group A: BMI < 23 kg/m2). Their height, body weight, waist and hip circumference, fasting plasms glucose (FPG), fasting plasma insulin (FINS), and serum leptin were examined at the baseline and 12 weeks after the therapy. Insulin senstivity index (ISI) was calculated [ISI = 1/(FPG x FINS)]. Multiple linear regression analysis and partial correlation were performed on serum leptin. RESULTS: The body weight, BMI, waist and hip circumference decreased significantly after the 12 week-treatment with sibutramine in group B (P < 0.01), but those indexes did not change after the treatment with placebo in group C (P > 0.05). The levels of leptin and FINS were higher (P < 0.01), but ISI was lower (P < 0.01) both in group B and C compared with those in group A at the baseline. The levels of serum leptin and FINS decreased (P < 0.01), and ISI increased significantly (P < 0.05) after the treatment with sibutramine in group B, while those indexes did not change after the treatment with placebo in group C. The most important factors to influence serum leptin level were listed as follows: sex > BMI > FINS > ISI (R2 = 0.661, F = 12.662, P < 0.01). The lep- tin was positively correlated with FINS (r = 0.597, P < 0.01) , but negatively correlated with ISI (r = -0.468, P < 0.01 ) after eliminating the effects of sex and BMI. Conclusion Leptin resistance and insulin resistance exist in obesity, and serum leptin is associated with IR. Treatment with sibutramine significantly reduces the body weight and leptin, increases insulin senstivity, and improves IR. PMID- 15804072 TI - [Clinical value of tumor supplied group of factor combined with CEA in diagnosing tuberculosis pleural effusion and malignant pleural effusion]. AB - OBJECTIVES: To determine the clinical value of tumor supplied group of factor (TSGF) combined with carcinoembryonic antigen (CEA) in diagnosing tukerculosis pleural effusion and malignant pleurul effusion. METHODS: TSGF and CEA were detected by ELISA in 14 patients with tuberculosis pleural effusion and 28 patients with malignant pleural effusion. RESULTS: The average levels of TSGF and CEA in patients with malignant pleural effusion were higher than those with tuberculosis pleural effusion. The diagnostic sensitivity of TSGF and CEA in the malignant pleural effusion was 67.5% and 57.5%, the specificity was 85.7% and 78.6%, and the agreement rate was 72.2% and 62.9% respectively; while the sensitivity, specificity and agreement rate of TSGF combined with CEA were 93%, 67.4%, and 88.9%. CONCLUSION: The sensitivity and agreement rate of TSGF combined with CEA are higher than those of either of the two methods. TSGF combined with CEA is important in differentiating tuberculosis and malignant pleural effusion. PMID- 15804073 TI - [Serum soluble CD40L level and its application in coronary heart disease]. AB - OBJECTIVE: To investigate the serum level of soluble CD40 ligand (sCD40L) in coronary heart disease and its relationship with serum lipid levels. METHODS: Enzyme-linked immunosorbent (EIA) was used to measure sCD40L in 129 patients with coronary heart disease (46 with acute myocardial infarction, 37 with unstable angina pectoris and the other 46 with stable angina pectoris) and 30 matched healthy controls. RESULTS: sCD40L level was significantly higher in patients with acute coronary syndrome (ACS) [(2. 99 +/- 2.19), (2.23 +/- 0.67) ng/ml in acute myocardial infarction and unstable angina pectoris group, respectively] than that of those with stable coronary heart disease and that of controls [(1.12 +/- 0.42) and (0.89 +/- 0.55) ng/ml, respectively] (P < 0.05). Serum sCD40L level was significantly positively related to ApoB100 (r = 0.159, P = 0.046), and negatively related to HDL-C (r = -0.192, P = 0.015). CONCLUSION: The levels of sCD40L increase in acute coronary syndrome, which may be related to the pathogenesis of ACS, and can be used as a potential marker of plaque instability. PMID- 15804074 TI - [Preservation of the entire mitral subvalvular apparatus during mitral valve replacement in patients with mitral stenosis]. AB - OBJECTIVE: To evaluate the surgical technique and the effect of preservation of the entire mitral subvalvular apparatus during mitral valve replacement (MVR) in patients with mitral stenosis. METHODS: MVR was performed on 56 patients with mitral stenosis. Of them, 11 had complete preservation of the entire mitral subvalvular apparatus (group A) with the technique of preservation of 'button shaped' transplantation, 25 had the preservation of the posterior leaflet only (group B), and the other 20 underwent the conventional MVR (group C). Pre- and postoperative hemodynamic parameters were measured to determine the left ventricular performance. Echocardiography was performed preoperatively, at the time of discharge, and 3 to approximately 6 months postoperatively to determine the dimensions and ejection function. RESULTS: Cardiac index, stroke volume index and left ventricular stroke work index postoperatively in group A were better than those in group B and C (P < 0.05). Echocardiographic measurements postoperatively showed the increase of LVESD, LVEDD in group B and C, but a better LVL and LVFS in group A (P < 0.05), and LVEF was more pronounced in group A (P < 0.05). There appeared a long cross-clamp time in group A, but the total CPB time was not significant among the three groups. CONCLUSION: The surgical skills of preservation with 'button-shaped' transplantation may be a practical choice for patients with mitral stenosis. The preservation of entire mitral subvalvular apparatus improves the left ventricular functions after MVR in patients with mitral valve stenosis. PMID- 15804075 TI - [Effect of glutathione pretreatment on lung ischemia-reperfusion injury during cardiopulmonary bypass]. AB - OBJECTIVE: To observe the clinical effect of glutathione (GSH) pretreatment on lung ischemia-reperfusion injury during cardiopulmonary bypass (CPB) and determine the mechanism. METHODS: Sixteen patients ready to replace heart valve(s) were randomly divided into the control and the treatment group. GSH diluted with normal saline solution was injected to the patients in the treatment group 36 h before the operation, while normal saline solution was injected to the patients in the control group. Malondiadehyde (MDA), Factor V III (FV III) and free hemoglobin (FHb) were detected in the left and right heart chamber at pre CPB 5, 10, and 15 minutes after the heart resuscitation. The differences between the left and right were recorded. Arterial blood from the radius was analyzed for PO2/FiO2 at pre-CPB, 10 min, 1 h and 6 h after the heart resuscitation. RESULTS: (1) The impairment of PO2/FiO2 in the treatment group was less than that in the controls. (2) MDA, FV III and FHb concentration in the left chamber were higher than those in the right. Increases of the parameters in the treatment group were lower than those in the controls. (3) The simple correlations were determined between MDA-FV III, FHb-MDA and FHb-FV III in both the control and the treatment groups (r = 0.774, 0.753 and 0.802), with statistical significance. CONCLUSION: GSH pretreatment can protect early lung ischemia-reperfusion injury during CPS, whose mechanism may be to decrease the leakage of FHb, and inhibit the free radicals reaction. PMID- 15804076 TI - [Different patterns of the left ventricle of heart in hypertensive patients]. AB - OBJECTIVE: To observe the clinical characteristics of different patterns of the left ventricule in hypertensive patients. METHODS: Echocardiography was used in 228 essential hyventricule of heart in hypertensive patients and 60 healthy individuals. RESULTS: In the hypertensive group, the rate of normal left ventricle, concentric remodeling, concentric hypertrophy and eccentric hypertrophy were 21.9%, 36.4%, 32.9% and 8.8%, respectively. The aortic root dimension and left atrial dimension were higher in the hypertensive group than those in the normal group (P < 0.05), especially for concentric hypertrophy and eccentric hypertrophy. Cardiac index and stroke volume index in eccentric hypertrophy were higher than those in the normal group and concentric remodeling (P < 0.05). Ejection fraction and fractional shortening in concentric remodeling were the highest and ejection fraction in eccentric hypertrophy was the lowest among all patterns (P < 0.05). E/A rate was lower in the hypertensive group than in the normal group (P < 0.05), especially for concentric hypertrophy and eccentric hypertrophy. The total peripheral resistance was the highest in concentric remodeling and the lowest in eccentric hypertrophy (P < 0.05). CONCLUSION: Different patterns in the left ventricle have their own characteristics and patients should be treated differently according to the patterns. PMID- 15804077 TI - [Cyclosporin A combined with dexamethasone in preventing and treating immune rejection after penetrating keratoplasty]. AB - OBJECTIVE: To evaluate the efficacy of topical 1% cyclosporin A (CsA), 0.1% dexamethasone or 1% CsA combined with 0.1% dexamethasone in preventing and treating immune rejection after penetrating keratoplasty (PKP). METHODS: Eighty six eyes from 86 PKP patients were randomly divided into 3 groups: (1) Thirty-one eyes were treated with 1% CsA and dexamethasone for 3 months. (2) Twenty-nine eyes were treated with 1% CsA for 3 months; (3) Twenty-six eyes were treated with 0.1% dexamethasone for 3 months. The rejected eyes of postoperation were given with the dexamethasone injection under conjunctiva and increased the frequency of CsA and dexamethasone eye drops. All patients were followed up for 1 to approximately 2 years. RESULTS: There was a statistical difference in the 3 groups in the postoperative immune rejection which occurred in 5 out of 29 (17.3% ) eyes treated with 1% CsA, 7 out of 26 (26.9%) treated with 0.1% dexamathasone, and 3 out of 31 (9.7%) with 1% CsA and dexamethasone. The immune rejection after PKP occurred in 15 eyes and 13 eyes were cured by sub-conjunctiva injection of dexamethasone combined with eye drops of 1% CsA and 0. 1% dexamethasone. CONCLUSION: The efficacy of CsA combined with dexamethasone topically is better than that of 1% CsA or 0.1% dexamethasone alone in preventing rejection episodes. It is effective to cure the graft rejection after PKP with sub-conjunctiva injection of dexamethasone combined with the eye drop of 1% CsA and 0.1% dexamethasone. PMID- 15804079 TI - [Percutaneous transcatheter closure for secundum atrial septal defect]. AB - OBJECTIVE: To evaluate the effect of transcatheter closure treatment for secundum atrial septal defect(ASD). METHODS: ASD was established by transthoracic echocardiogram (TTE) and transvenous cardiac catheterization. A balloon catheter was inserted at ASD and was inflated to measure the ASD stretched diameter. Then, a long sheath was inserted to the left atrium (LA) from the right atrium( RA), Thereafter a proper auto-expanded atrial septal occluder (ASO) was inserted to the tip of the sheath. Under the fluoroscopic and TTE guidance, the ASO was expanded. Once the position of ASO was optimal, it was released. RESULTS: From May 1999 to July 2002, 32 patients were treated with this method. Among them, 28 patients (87.5%) obtained satisfactory results. At the end of the operation, TTE could not find out any residual shunt. Before discharging the patients, TTE showed the diameter of the right atrium and the right ventricle were decreased from (39.4 +/- 5.8) mm to (33.7 +/- 4.2) mm and from (45.3 +/- 6.8) mm to (37.1 +/- 4.5) mm respectively (P < 0.001 for both). The operation failed in 4 patients (12.5%). Two patients did not get proper ASO for them in the early period of our work. ASO slipped off from the defects in another 2 patients during the operation. There were no other complications. CONCLUSION: Tanscatheter closure is suitable for most patients with ASD. It is safe and less invasive with few complications. PMID- 15804078 TI - [Effects of nitric oxide on form-deprivation myopia in chicks]. AB - OBJECTIVE: To construct a chick model of form-deprivation myopia (FDM) to investigate the effect of nitric oxide (NO) on FDM. METHODS: FDM was induced in 60 one-day white leghorn chicks. They were divided into group A and B. One eye was covered randomly, and the other served as control. In each week of the experimental period, the retinoscopic refraction was performed, and the axial length was measured with A-mode ultrasound. The NO concentration of retina and choroid from the control and the experimental eyes was measured using NO detection kit. Change in the expression of iNOS mRNA was measured by RT-PCR. The activity of iNOS was analyzed with immunohistochemistry. RESULTS: Form deprivation eyes were significantly more myopic than the controls, and the axial length of occluded eyes was significantly longer than that of the controls. Unocclusion attenuated the development of myopia and inhibited the axial elongation. Form-deprivation reduced the NO concentration ,the immunoreactivity of iNOS, and the expression of iNOS mRNA. The gradual restoration of normal vision was found after uncovering the eyes. But the expression of iNOS mRNA was significantly higher than that of the controls at the first week after the unocclusion. CONCLUSION: Form-deprivation can construct an animal model of myopia. FDM is reversible before the chicks were mature. NO may be involved in the form of FDM. PMID- 15804080 TI - [Serum levels of antibodies for IgG, IgA, and IgM against the fungi antigen in psoriasis vulgaris]. AB - OBJECTIVE: To detect the serum levels of antibodies against Malassezia further and Candida albicans in patients with psoriasis vulgaris (PV), and to investigate the relationship between Malassezia furfur or Candida albicans and pathogenesis of PV. METHODS: The serum levels of antibodies for IgG, IgA, and IgM against the whole cell antigen Wag and soluble antigen (Sag) of Malassezia fufur and Candida albicans in 40 PV patients and 20 healthy controls were detected by indirect ELISA, RESULTS: The serum levels of IgG against Wag of Malassezia furfur and Candida albicans were significantly higher in PV patients than those in the healthy controls (P < 0.01, P < 0.05, respectively), The serum levels of IgM against Sag of Malassezia furfur and Candida albican, IgA against Sag of Candida albicans were significantly lower in PV patients than those in the healthy controls (P < 0.05). CONCLUSION: There are some humoral immunity abnormalties in PV patients. Malassezia furfur and Candida albicans may be related to the pathogenesis of PV. PMID- 15804082 TI - [Clinical analysis of intrahepatic cholestasis during pregnancy in 150 patients]. AB - OBJECTIVE: To explore the effect of intrahepatic cholestasis of pregnancy (ICP) on the mother and fetus, and to assess the clinical value of nonstress test. METHODS: The symptoms, biochemical examinations, fetal monitoring, and pregnant outcomes of 150 patients with ICP were analyzed retrospectively in our hospital. RESULTS: There was no linear relation among serum cholanic acid, glutamic-pyruvic transaminase, total bilirubin, and gestational age when skin pruritus of pregnant women appeared, but there was positive correlation among the neonate status, meconium-stained amniotic fluid, and the onset time of pruritus. There was no correlation between the amount of postpartum hemorrhage and the newborn Apgar scores. No significant difference was found in perinatal prognosis The appearance of skin between normal and abnormal outcomes in the nonstress test. CONCLUSION: pruritus can be a sign of the state of ICP patients. To carefully monitor and comprehensively and continuously estimate the fetal state, and to terminate the pregnancy at proper time are very important to improve the prognosis of perinatal infants. PMID- 15804081 TI - [Etiologic analysis of 1137 inpatients of chronic renal failure]. AB - OBJECTIVE: To investigate the main causes of chronic renal failure (CRF) and the differences in how subpopulation is affected by these causes, and to determine the morbidity of anemia and hypertension, and the relation between age and gender in CRF patients. METHODS: We chose 1137 inpatients diagnosed as CRF in the Department of Nephropathy of 2nd Xiangya Hospital of Center South University from January 1996 to March 2002. We collected the information on name, gender, age, admission number, diagnosis, blood pressure, blood routine, renal function, and creatinine clearance rate. RESULTS: Chronic glomerulonephitied accounted for 56.4% as the leading cause, followed by hypertensive nephropathy (14.3%), obstructive nephropathy (7.2%), and diabetic nephropath (6.7%). Gender influenced the incidence of some diseases. Most patients were companied with anemia (90.7%) and hypertension (72.6%). CONCLUSION: The main causes of CRF include chronic glomerulonephitis, hypertensive nephropathy, obstructive nephropathy, and diabetic nephropath. PMID- 15804083 TI - [Meconium-stained amniotic fluid and intra-amniotic infection]. AB - OBJECTIVE: To explore the relationship between meconium-stained amniotic fluid and Fifty-six women of cesarean section with intact membrane and intra-amniotic infection. METHODS: without parturient were divided into 3 groups according to the property of amniotic fluid (no meconium, I to approximately II degree meconium stainedness, and III degree mecomium stainedness). The content of interleukin-6 in amniotic fluid was measured with ELISA. The infiltration of inflammatory cells in the placenta and its membrane was determined by the pathological diagnosis. The neonatal Apgar score and puerperial infection after the surgery were analyzed. RESULTS: There were no significant differences in the content of IL-6 in amniotic fluid and in the infiltration of inflammatory cells among the 3 groups. But the rate of neonatal asphyxia in the meconium-stained cases was significantly higher than that without meconium. CONCLUSION: Meconium stained amniotic fluid is a marker of fetal distress, but it is not related to intra-amniotic infection. PMID- 15804084 TI - [Immunohistochemical study of atriomyxoma]. PMID- 15804085 TI - [Effect of rocuronium on hemodynamic in patients undergoing cardiac valve replacement]. PMID- 15804086 TI - [Significance and plasma level of PAI-1 on the angiographic severity of coronary lesions and coronary event]. PMID- 15804087 TI - [Clinical analysis of cervical lymph node metastasis as primary manifestation of oral cancer]. PMID- 15804088 TI - [Fourteen cases of primary malignant lymphoma of bone]. PMID- 15804089 TI - [Fundamental medicine: the role of insulin in providing energy for the biological function of locomotion (a review of literature)]. PMID- 15804090 TI - [Lipid peroxidation in neurological pathology in children]. AB - The intensity of lipid peroxidation (LPO) was studied by the content of the listed below in erythrocytes of children with neurological pathology: diene conjugates, trienketones and LPO products. Intensified LPO processes were registered in children with neurology and primarily in those with cerebral spastic infantile paralysis (CSIP). The detected changes in the LPO products of erythrocyte membranes depended on a disease variation on an intellectual development of sick child. The LPO activation affects possibly the development (lesion) of craniocerebral innervation in patients. Changed LPO parameters were found in examined sick children to be most closely related with the development of their motor skills and with the formation of the bone-muscle system, thus, an essentially higher content of LPO products was registered in erythrocytes of children with a reduced muscular activity (in those not capable of sitting, standing or walking). Considering the above stated and with respect to the deficit of Vitamin E registered in such patients, children with CSIP or with impending CSIP need, obviously a prolonged membrane-stabilizing therapy by antioxidants and essential phospholipids including traps of radicals, which should cut the number of children with severe CSIP and with residual perinatal lesions of the central nervous system. PMID- 15804091 TI - [Acoustic (ultrasound) determination of concentration of whole protein and protein fractions in blood serum]. AB - A new method is suggested for determining the concentration of whole protein and the distribution of protein fractions in blood serum by means of an ultrasound device. The acoustic method is based on measuring the proliferation rate and absorption of the ultrasound signal in the examined serum. The findings of the method were compared with those obtained by routine tools in examining both blood sera of patients and controls. A high correlation factor was registered for the biuret method in the determination of the concentration of whole protein (R = 0.95; p < 0.05). It was demonstrated as possible to use the discussed method as a screening tool in evaluating the distribution of protein fractions. The examination findings are generated by the instrument as in the electrophoretic examination. The method is simple in use. No reagents are needed. PMID- 15804092 TI - [Endogenous intoxication in patients with stomach cancer observed immediately after surgery]. AB - Severe endogenous intoxication (El) can trigger immediately after surgery many complications right up to multiple organ failure. The evaluation of El according to a degree of toxemia is most objective up to now. The purpose of the case study was to follow up the dynamics of the main El laboratory indices and to evaluate the correlation between them in patients with stomach cancer during the early postoperative period. The El parameters were determined in 2 hours before surgery as well as on days 1, 3, 5, 7 and 14 after surgery. The below indices were determined in blood of patients: toxicity index (TI), count of mean weight molecules (MWM), leukocytic intoxication index (LII) and total count of leukocytes (TCL). The study showed that, when patients with stomach cancer are admitted for surgical treatment, they have pronounced El, which intensifies even more postoperatively. The investigated laboratory indices are independently significant in the evaluation of the El degree. A reliable but faint correlation was found between the parameters of MWM and IT, and MWM and LII. TCL does not correlate reliably with other studied El parameters except for LII. The full fledged evaluation of the El nature must be used in choosing purposefully detoxication methods with the aim of promoting the quality of postoperative treatment of patients with stomach cancer. PMID- 15804093 TI - [Mean molecule peptides of plasma and blood serum]. AB - The concentration of mean molecule peptides is by far less in serum versus plasma. The concentration of mean molecule peptides in serum can go down, in endotoxicosis degrees 3-4, to normal values, whereas it can exceed 3-4-fold the normal value in plasma. The most pronounced reduction of peptides' concentration in serum was registered for the fraction with the molecular weight ranging from 800 to 3000 D. PMID- 15804094 TI - [Changing parameters of microcirculation, coagulogram and morphometry of the nucleolar organizers zone in the epidermis of patients with psoriasis]. AB - Findings of morphology, coagulogram and microcirculation in the derma, i.e. pathogenetically significant parameters in the onset and progression of prosiasis, were analyzed for 100 patients with psoriasis. Essential changes were detected, which apparently bring about lesions in the histophysiology of tissue homeostasis. PMID- 15804095 TI - [Human mycoplasma and mycoplasmal infection (lecture, Part 1)]. PMID- 15804096 TI - [The activity of glutathionereductase in neutrophils of patients with mucoviscidosis]. PMID- 15804097 TI - [Time of analysis and final parameters of peripheral blood]. AB - The blood parameters are influenced by the balanced state of all blood elements preserved during a certain time interval. A comparison of blood-test findings obtained during the initial two hours and in 20-24 hours after sampling and storage in presence of a stabilizer denoted a decreased quantity of leukocytes and erythrocytes by 2.57.7% and 12-22%, respectively. At the same time, the leukocytic formula and ESR remained unchanged. PMID- 15804098 TI - [The specificity of synthesis of cytokines in patients with coxarthrosis in hip replacement]. PMID- 15804099 TI - [A modified test-system for the detection of IgM antibodies to the core-antigen of viral hepatitis B in human blood serum_ELISA-anti HBc-M-screen]. PMID- 15804100 TI - [Laboratory diagnostics of urogenital clamidiosis]. AB - Laboratory diagnostic tools of urogenital clamidiosis--PCR, ELISA (IgG and IgM) and direct immunofluorescence (DIF)--were comparatively analyzed. The positive PCR result was checked by another PCR test with a different primer; 5 false positive responses were registered (specificity 99.6%). As against PCR, the sensitivity of ELISA made 53%, its specificity -75.5%, the diagnostic value of positive result -58%, the diagnostic value of negative result -71.6% and the diagnostics accuracy -66.7%. The respective DIF parameters were as follows: 36%, 90%, 81.5%, 54.2% and 60.9%. A high rate of detection (above 90%) of the conditionally pathogenic microflora associated with Chlamydia trachomatis (above 110 microbe cells/ml) was pointed out. Hardnerelli and ureaplasms were more often found in female smears, staphylococci and enterococci--in male sperm. It is underlined as important to hold complex examinations for Chlamidia (PCR, ELISA and DIC as an additional test) combined with bacteriological quantification of the conditionally pathogenic microflora and determination of its resistance to antibiotics. PMID- 15804101 TI - [The results of training laboratory experts from Central Asia in ensuring the quality control in the HIV diagnostics]. PMID- 15804102 TI - [PCR-based diagnostics of community-acquired pneumonia of mycoplasmal and Chlamydia etiologies]. AB - Two hundred and ninety-two patients with pneumonia evidenced by clinical, X-ray and laboratory findings were examined for the purpose of studying the etiological significance of Mycoplasma pneumoniae and Chlamydophila pneumoniae--atypical pathogens; the patients, members of closed bodies, were admitted to the hospital for as long as one year. PCR was used to examine the sputum of patients who were at exacerbation. It was established that the mentioned pathogens prevailed in autumn and winter. It can be concluded, on the basis of a PCR-based monitoring of the atypical pathogens, that the significance of the atypical pathogens within the etiological structure of pneumonia acquired in isolated collective by young persons has been on the rise. PMID- 15804103 TI - [Determination of the bactericidal activity of blood serum by recombinant luminescence strains of Escherichia coli]. AB - It has been experimentally proven as possible to use recombinant strains of Escherichia coli carrying the genes of luminescence of natural fluorescent bacteria in evaluating the bactericidal activity of blood serum (BABS). The shaping fluorescence inhibition was shown to be in proportion to the developing bactericidal effect in respect to the luminescent strains of Escherichia coli, which depended on a used concentration of blood serum and time of contact. The list of E. coli recombinant strains, which fit in most of all with the evaluation of BABS, was drawn. The values of the bactericidal activity of blood serum determined by the routine bacteriological and nephelometric methods were demonstrated to coincide with those determined by the worked-out bio-luminescent method. PMID- 15804104 TI - [AIDS vaccines and immunotherapy of infertility]. AB - The development of prophylactic AIDS vaccines and immunotherapeutic approaches such as therapeutic AIDS vaccines would greatly benefit from acquired experience in reproductive immunology field relating to pregnancy and infertility. Certain immune and pathological aspects of HIV infection are closely related to problems faced by obstetricians, embryologists, gynecologists, and andrologists. This review attempts to bring together the recent advances in AIDS field with progress made in the physiology and pathology of reproduction in humans. The insight gained in reproductive biology research, particularly in the area of immunotherapy of recurrent spontaneous abortions, can help to develop better strategies for the treatment and prevention of HIV infection. PMID- 15804105 TI - [On the significance of RNA in human sperm]. AB - Our understanding of the molecular complexities of human spermatozoa continues to be rather rudimentary. This includes the unique packaging and structure of the sperm genome along with its paternally derived RNAs in preparation for their delivery to the egg. Both highlight the inadequacy in our understanding of how these genetic factors contribute to a healthy child. As a means to address this deficiency it was recently shown that human spermatozoal RNA isolated from the ejaculate can be used to transcriptionally profile the male gamete. It is fully expected that a genetic fingerprint characteristic of the normal fertile spermatozoon can be identified and that deviations from this profile can be detected. PMID- 15804106 TI - [Establishment and application of the approach to detecting two biovars of Ureaplasma urealyticum in human semen]. AB - OBJECTIVE: To establish the approach to detecting two biovars of Ureaplasma urealyticum (Uu) in human semen and to investigate the relationship between the two biovars of Uu infection and the quality of human semen. METHODS: Based on the 16S-23S rRNA intergenic spacer region, three pairs of primers were designed, the species specific primer and two biovars primers (Parvo primer and T960 primer). The two biovars of Uu were detected in the semen from 949 men by semen culture and PCR assay. Meanwhile, semen routine analyses were performed. RESULTS: In the 949 subjects, 199 were Uu positive both in Uu liquid culture and PCR assay (199/949, 21.1%), of which 136 (136/199, 68.3%) were Parvo biovar, 54 (54/199, 27.1%) T960 biovar, and 9 (9/199, 4.5%) both Parvo and T960 biovars. Compared with the Parvo and the negative groups, human sperm viability was significantly decreased (P < 0.05 ) in the Uu T960 infection group. The difference of sperm motility and density had no statistic significance. CONCLUSION: A significant correlation has been found between Uu T960 biovar infection and human sperm viability PMID- 15804107 TI - [Influence of Candida albicans on the motility and ultrastructure of human spermatozoa in vitro]. AB - OBJECTIVE: To explore the influence of Candida albicans (Ca) on the motility and ultrastructure of human spermatozoa and its possible mechanism. METHODS: Semen samples obtained from 10 healthy volunteers by masturbation were prepared by the swim-up technique and sperm density to 40 x 10(6)/ml. The samples were then inoculated at 37 degrees C with different concentrations of a uropathogenic strain of Ca isolated from an outpatient, with initial fungi/spermatozoa ratios varying among 1:1 (Group A), 1:10 (Group B), 1:100 (Group C), 1:1000(Group D), and 1:10,000 (Group E). And Group F containing Ham's F-10 only was found as the negative control. Motion parameters were analysed by computer-aided sperm analyzer (CASA) at 0 hour, 1 hour, 2 hours and 4 hours respectively. Modalities of spermatozoa and possible adherence and/or agglutination were observed under the light microscope. Finally, all the samples were studied by transmission electron microscopy. RESULTS: Distinct adhesion of spermatozoa to Ca and agglutination were noticed. In all the motion parameters, progressive motility was affected most and dependent upon incubation time and bacterial concentration. Progressive motility showed a significant difference between Group A and the control (P < 0.01). With the prolongation of incubation time, other parameters were showing more and more differences. Analysis by electron microscopy revealed multiple ultrastructural damages. CONCLUSION: Ca significantly inhibits human sperm motility and decreases sperm viability in vitro. Its mechanism is possibly related to Ca's adhesion to human spermatozoa and the impairment inflicted by Ca to sperm ultrastructure. PMID- 15804108 TI - [Analysis of Yq microdeletions in idiopathic infertile males with azoospermia and oligospermia in Shaanxi Province]. AB - OBJECTIVE: To evaluate the frequency of microdeletions in the long arm of Y chromosome of idiopathic infertile males with azoospermia and oligospermia in Shaanxi province in China and to investigate the relevance of sperm count to Y microdeletion frequencies. METHODS: According to the sequence of sequence-tagged sits (STS) AZFa, AZFb, AZFc and SRY, 4 of the azoospermic factor regions on Y chromosome long-term supplied by GenBank, 5 sets of primers were synthesized. The Y microdeletions in AZF regions were screened by polymerase chain reaction (PCR) in 64 idiopathic cases of azoospermia and oligospermia and 20 men of known fertility. RESULTS: No microdeletion was detected in the 20 normospermic subjects. Deletion of the AZFc/DAZ was detected in 11 individuals and one patient had both AZFb and AZFc deletion; no deletion of AZFa and SRY region was found. The frequency of Y microdeletions in the subgroups with different sperm count showed the highest value among azoospermic men (3 cases, 21.4%). The percentage progressively decreased with the deletion frequency (20.0%, 17.9% and 8.3%) in the subgroups with sperm counts of < 1 x 10(6)/ml, < (1-5) x 10(6)/ml and < (1 to approximately 10) x 10(6)/ml, respectively. CONCLUSION: Y chromosome microdeletions are specifically associated with severe spermatogenic failure. The rate of deletion involving AZF region of the Y-chromosome is higher in infertile men with azoospermia and oligospermia. PCR amplification of AZF locus is useful for the diagnosis of microdeletions in the Y-chromosome. PMID- 15804109 TI - [Surgical management of male infertility beyond assisted reproductive technology]. AB - OBJECTIVE: To investigate the role of urological surgery for male infertility which is beyond the help of assisted reproductive technology (ART). METHODS: We retrospectively reviewed the records of 7 male infertility patients who, having failed to respond to ART, underwent urological surgery from 1999 to 2003. Of the 7 cases, 4 were varicocele, 2 were post-bilateral vasectomy and 1 was bilateral chronic epididymitis. For the 7 patients, 6 times we performed in vitro fertilization-embryo transfer (IVF-ET) and 5 times we attempted intracytoplasmic sperm injection (ICSI) , but all failed. After that, we resorted to surgical approaches, ligation of internal spermatic veins in 4 cases, bilateral vasovasostomy in 2, and unilateral epididymovasostomy in 1. RESULTS: After surgical intervention, 2 cases fathered children, 1 achieved pregnancy and 3 cases improved in the quality and quantity of seminal fluid; only 1 failed to respond to the therapy. CONCLUSION: In the treatment of male infertility, priority should be given to surgical intervention and, in case of failure, assisted reproductive technology could be considered. PMID- 15804110 TI - [Bi-planar transrectal ultrasonographic study on the diagnosis of hyperplasia prostate gland]. AB - OBJECTIVE: To discuss the value of grey scale ultrasonography and color Doppler of bi-planar transrectal ultrasound on the diagnosis of hyperplasia prostate gland (HPG). METHODS: One hundred and ninety-two patients with HPG and 50 control subjects underwent abdominal ultrasound and TRUS examinations. The width (W), depth (D) and length (L) of the prostates and transitional zones were respectively measured. Each transitional zone index (TZI), i.e., the ratio of the transitional zone volume to the total prostate volume was calculated. Meanwhile, we observed the blood flow distribution in the prostates, calculated the parameters of hemodynamics such as peak systolic velocity (PSV), resistance index (RI) and pulse index (PI), and analyzed their correlations with TZI. RESULTS: The revealing rates of cystic formation, calcium calculus and ejaculatory cyst and PSV, RI of arterial vessels in HPG were all significantly higher in the HPG patients than in the control subjects, and the revealing rates of all the above pathological changes but calcium calculus by TRUS were significantly higher than by abdominal examination. TZI of HPG was positively correlated with PSV, RI and PI. CONCLUSION: Bi-planar TRUS can clearly display the normal and pathological structure of the prostate. And PSV, RI and PI may aid in the diagnosis of HPG and the assessment of the degree of prostatic proliferation. PMID- 15804111 TI - [High intensity focused ultrasound combined with endocrine therapy in treating prostate cancer]. AB - OBJECTIVE: To evaluate the clinical efficacy of high intensity focused ultrasound (HIFU) combined with endocrine therapy in the treatment of patients with prostate cancer. METHODS: Twenty patients with prostate cancer were treated with extracorporeal HIFU device( model FEP-BY01 ) and androgen ablation, of whom 15 received orchiectomy and 5 LHRH-a. Fourteen patients of the total number were given flutamide in addition to castration. RESULTS: The mean follow-up was 13.5 months (ranging 6 to approximately 30). Before and after the treatment, the prostate volume, prostate specific antigen (PSA), international prostate symptom score (IPSS) and maximal flow rate (Qmax) of the patients were (36.4 +/- 16.2) ml and (20.6 +/- 11.8) ml (P < 0.05), (32.2 +/- 10.4) ng/ml and (2.4 +/- 0.8) ng/ml (P < 0.01), 20. 5 +/- 6.5 and 13.6 +/- 7.5 (P < 0.05), (10.6 +/- 6.3) ml/s and (14.2 +/- 4.6) ml/s (P < 0.05), respectively. Mild hematuria and pain were noted in 5 and 8 patients respectively, and 1 patient underwent internal urethrotomy with a cold knife because of urethral stricture. er, with minimal complications. CONCLUSION: HIFU combined with endocrine therapy is effective in the treatment of prostate canc- PMID- 15804112 TI - [Reconstruction of the remnant penis: a 52-case report]. AB - OBJECTIVE: To explore the effects of the combined method of abdominal axial flap transposition and penile elongation for the treatment of the remnant penis. METHODS: Fifty-two cases of the remnant penis treated with the combined method from 1984 April to February 2004 were analyzed retrospectively. Follow-up ranged from 0.5 to 20 years postoperatively. RESULTS: The lengths (both in normal and erectile conditions) and the circumferences of the penis gained after operation were (5.6 +/- 1.4) cm, (6.8 +/- 2.5 cm and (6.9 +/- 2.3) cm respectively. The recovery rates of the sensory function were 94.2% and 100% in the glans (immediately and 3 months after operation) and 32.7%, 51.9% and 75% in the flap area (3, 6 and 12 months postoperatively). The two-point distinguishing sense in the glans and the flap area was (5.1 +/- 0.9) mm and(7.9 +/- 1.3) mm 5 years after operation. Early complications included distant flap necrosis (3 cases), disruption of the wound (2 cases), part necrosis of the skin graft in the abdominal wall (2 cases) and poor contours occurred in 4 cases in the later period because of the thickness of the flaps. All of them were corrected with satisfactory results. CONCLUSION: The combined method of abdominal axial flap transposition and penile elongation was recommendable for the treatment of the remnant penis because of its positive effects and less complications. PMID- 15804113 TI - [Analysis of cytokines (IL-2, IL-8, IL-10) in the expressed prostatic secretions of chronic prostatitis]. AB - OBJECTIVE: To investigate the levels of immunoregulatory cytokine IL-2, pro inflammatory cytokine IL-8 and anti-inflammatory cytokine IL-10 in the expressed prostatic secretions (EPS) of chronic prostatitis (CP) patients and to evaluate the significance of the cytokines to the pathogenesis and diagnosis of CP. METHODS: IL-2, IL-8 and IL-10 levels were measured in the EPS of 31 CP patients and 10 normal controls by enzyme-linked immune sandwich assay (ELISA). Urine was cultured and EPS studied according to the 2-glass test. NIH-CPSI (NIH-chronic prostatitis symptom index) was performed in every patient. The cases of CP were divided into 3 types: II (n=5), IIIA (n=13) and IIIB (n=13) according to NIH. RESULTS: The IL-8 levels in CP patients were significantly higher (P < 0.05) while the IL-2 and IL-10 levels significantly lower than those in the controls (both P < 0.05). There was no statistically significant difference between the cytokine levels in II CP and in IIIA CP (P > 0.05). The IL-8 levels in IIIB CP were significantly lower than those in both II CP and IIIA CP (both P < 0.05). The IL-8 levels were correlated directly with WBC count (r = 0.663, P < 0.05) , and inversely with IL-10 levels (r = -0.503, P < 0.05), but there was no correlation between NIH-CPSI scores and cytokines levels (P > 0.05). CONCLUSION: Cytokines may play an important role in pathogenesis of prostatitis. The results indicate that the expression of cytokines (IL-2, IL-8, IL-10) in EPS can serve as a valuable marker for the diagnosis of CP. PMID- 15804114 TI - [Influence of cryoprotectant with glycerol and freezing-thawing procedure on the motility of human sperm]. AB - OBJECTIVE: To investigate the effects of cryoprotectant with glycerol and the freezing-thawing procedure on the motility of METHODS: The motion characteristics of human sperm from 18 selected specimens were assessed by computer-assisted human sperm. sperm analysis before and after adding hyper-osmolarity cryoprotectant with glycerol and the freeze-thaw procedure, and the data were evaluated in pairs. RESULTS: The adding of cryoprotectant caused an increase in the proportion of rapid linear motile sperms (P < 0.05) and sperm velocities, including VCL, VSL and VAP (P < 0.005). But no changes were observed in the proportion of progressively motile sperms and sperm motility. Compared with the data from pre-frozen samples, velocities of post-thawed sperms and the percentage of motile sperms in each grade significantly declined (P < or = 0.01), so did ALH, while WOB, LIN and STR remained unchanged (P > 0.05). Significant differences in ALH, WOB, LIN and STR were observed only in comparison between the post-thawed and pre-treated samples. CONCLUSION: The number of Grade a sperms and sperm velocity increased after adding hyper-osmolarity cryoprotectant with glycerol. The sperm motile potential was impaired and even entirely destroyed in some cases by cryodamage. The influences on the sperm motion were differently induced in freezing-thawing procedures. PMID- 15804115 TI - [Value of preoperative urodynamics to the prognosis of transurethral prostate resection for benign prostatic hyperplasia]. AB - OBJECTIVE: To establish the value of preoperative urodynamics to the outcome prediction of transurethral prostate resection (TURP) for benign prostatic hyperplasia (BPH). METHODS: One hundred and sixty patients with BPH underwent urodynamic tests. Preoperative IPSS (International Prostate Symptom Score) and QOL (Quality of Life) were compared with those 8 to approximately 11 months after operation. RESULTS: The parameters of urodynamic tests (max. free flow, detrusor pressure at max. flow, Schafer grade, Abrams-Griffiths No, urethral resistance factor, cystometric capacity, effective capacity). IPSS and QOL were improved after operation (P < 0.001). And all the relative coefficients of linear dependence analysis, IPSS, QOL and max. free flow, detrusor pressure at max. flow, Schafer grade, Abrams-Griffiths value, urethral resistance factor, cystometric capacity, and effective capacity conducted postoperatively, were statistically significant. CONCLUSION: Preoperative urodynamics of transurethral prostate resection for benign prostatic hyperplasia may provide indication for operation and predict postoperative improvement of symptoms. Preoperative urodynamics should be considered as a routine examination. PMID- 15804116 TI - [Successful treatment of high-flow priapism by selective arteriographic embolization of the pudendal artery]. AB - OBJECTIVE: To evaluate the diagnosis and treatment of high-flow priapism (HFP). METHODS: Four cases of HFP following blunt trauma to the penis or perinea underwent diagnostic examination by colour-flow Doppler ultrasound and/or superselective pudendal arteriography, which revealed bilateral arteriocorporal fistula in 1 case and monolateral in the other 3. Penile detumescence was obtained in 2 cases by superselective bilateral/monolateral arteriographic embolization of the pudendal artery with absorbable gelatin RESULTS: The former 2 cases effected an immediate recovery of the sponge, while the other 2 cases received conservative treatment. Erectile function, able to perform normal sexual intercourse in approximately 2 months. But in the latter 2, follow-up revealed unsatisfactory potency. CONCLUSION: Superselective arteriographic embolization with absorbable gelatin sponge can provide a safe, selective and effective treatment for HFP patients. PMID- 15804117 TI - [Effects of heating on the epithelia of benign prostate hyperplasia in vitro]. AB - OBJECTIVE: To investigate the biological changes of the epithelia of benign prostate hyperplasia under heating at different temperatures and at different points of time after heating. METHODS: Cell morphology, MTT, flow cytometer, and the immunocytochemical method for detecting heat shock protein 70 and prostate specific antigen were used to observe the effect of heating on the primarily cultured epithelia of benign prostate hyperplasia at 45 degrees C and 60 degrees C and at 1 hour and 12 hours after heating. RESULTS: Heating at 45 degrees C for 15 min resulted in apoptosis, and at 60 degrees C, necrosis in most of the cells. The inhibiting effect of heating on the growth of cells was observed, more significant in the 60 degrees C group than in the 45 degrees C group. The cell phase arrest induced by heat, mainly G0/G1 arrest, was more significant in the 12 h group than in the 1 h group. Heating up-regulated the expressions of heat shock protein 70 and prostate specific antigen in cells. CONCLUSION: Heating can induce apoptosis, necrosis and growth suppression of the epithelia of benign prostate hyperplasia. Its process and mechanism are correlated with the cell phase arrest and the up-regulation of the expressions of heat shock protein 70 and prostate specific antigen. PMID- 15804118 TI - [Primary artery erectile dysfunction: one case report]. AB - OBJECTIVE: To evaluate the relationship between the deformation of penile artery and the primary artery erectile dysfunction, and to improve the treatment and diagnosis of primary artery erectile dysfunction. METHODS: One case of primary artery erectile dysfunction was presented with its primary clinic data. RESULTS: The dorsal artery of the penis was thin and the bilateral penile arteries were lacking by arteriography. The implantation of a penile prosthesis significantly improved the patient's erectile function. CONCLUSION: The primary artery erectile dysfunction is a relatively rare disease. The possibility of primary artery erectile dysfunction should be kept in mind. Penile prosthesis implantation is an effective means for the treatment of primary artery erectile dysfunction. PMID- 15804119 TI - [Penile vibratory stimulation and electroejaculation in the treatment of ejaculatory dysfunction]. AB - The fertility potential of infertile men can be enhanced to a great extent by the application of assisted reproduction techniques such as intrauterine insemination or in-vitro fertilization with or without intracytoplasmic sperm injection, but how to obtain semen from men with ejaculatory dysfunction remains a problem. The development and refinement of penile vibratory stimulation (PVS) and electroejaculation (EEJ) have significantly brightened the prospects for the treatment of ejaculatory dysfunction. Because vibratory stimulation is non invasive and easy to perform, and needs no anaesthesia, it is preferred by most of the patients to EEJ, and recommended to be the first choice of treatment for ejaculatory dysfunction. Approximately 80% of all ejaculatory dysfunction men with an intact ejaculatory reflex arc (above T10 ) can obtain antegrade ejaculation by PVS. Any condition which affects the ejaculatory mechanism of the central and/or peripheral nervous system including surgical nerve injury may be treated successfully by EEJ. The purpose of this review is to present the current understanding of PVS and EEJ procedures and their clinical use in men with ejaculatory dysfunction. PMID- 15804120 TI - [Safety of Sildenafil in men with cardiovascular diseases]. AB - Sildenafil is widely used to treat erectile dysfunction (ED). Because many patients with ED have cardiovascular diseases, doctors and patients are concerned over the safety of Sildenafil in patients with cardiovascular diseases. The review focuses on the safety of sildenafil for ED patients with cardiovascular diseases. PMID- 15804121 TI - [Application of Yao medicine in andrology]. AB - Yao medicine, with its long history and peculiar characteristics, has developed in the long practice of the Yao peoples fight against diseases. The past decade has seen frequent reports on the unique theories and techniques of Yao medicine in the diagnosis and treatment of andrological diseases. This paper reviews the original experiences of Yao medicine in the treatment of prostatitis, male infertility, male erectile dysfunction, and so on. PMID- 15804122 TI - [Clinical research on Linbiqing decoction for the treatment of chronic prostatitis following sexually transmitted disease]. AB - OBJECTIVE: To study the therapeutic effect of Linbiqing decoction on chronic prostatitis following sexually transmitted disease (CPFSTD). METHODS: Linbiqing decotion was given orally to 36 patients with CPFSTD, and the therapeutic effet was assessed by observing the clinical symptoms and measuring the objective indexes before and after the treatment. RESULTS: Four to eight weeks after the drug administration, the overall effectivity rate was 72.22%, and the scores of NIH-CPSI were decreased significantly. CONCLUSION: Linbiqing decoction has definite therapeutic effect on CPFSTD, and can be considered as an effective means for the treatment of the disease. PMID- 15804123 TI - [Guidelines and rehabilitation--examples from the EbM Symposium 2004]. PMID- 15804124 TI - [Clinical practice guidelines in Germany, 1994 to 2004. From guideline methodology towards guideline implementation]. AB - The role of clinical practice guidelines (CPG) as a tool for continuous medical education (CME), and quality management in health care is now widely accepted in Germany. Since the 90ies, the physicians' professional associations as well as health care authorities and parliament have been introducing several incentives and regulations in order to promote the use of evidence based CPG. In the past German CPG agencies have been focussing their work on developing and optimising methodological CPG standards. Future CPG activities should address much more other key factors for the success of CPGs, such as medical relevance, practicability, effective dissemination, and implementation. The article describes the process of guideline adaptation by regional physician audit groups as a tool for CPG implementation. PMID- 15804125 TI - [Guidelines and autonomy of physician decision. A critical survey under ethical aspects]. AB - The convergence of Diagnosis Related Groups (DRG) and Evidence-based Medicine (EbM) in the present German Health Care System Reform (GMG) leads, due to time constraints, to a misuse of EbM as cook-book medicine. The communication level of physicians has already shifted to abstract discussions of Randomized Controlled Trials (RCT) and guidelines, inducing per sea preference for pre-defined clinical decisions. The inconsistencies of evidence definitions, together with the overwhelming number of guidelines and providers thereof, burden medical reasoning with superfluous complexity, and contributing to the development of cook-book mentality. Without comprehensive trial registration of all RCTs before the start of a study, a publication bias will continue to exist. In the clinical practice, it is necessary to reduce the impact of study results and guidelines to a degree that remains compatible with patient interests and economy. In this field, prospective controlled outcome registries may provide a control for over- and faulty regulation. Clinical professionalism is characterized in parallel by a critical implementation of EbM and a necessary degree of independent reasoning. PMID- 15804126 TI - [Evidence-based rehabilitation and health care in view of social law]. AB - The explicit requirement for rehabilitation to be based on sound scientific evidence is only sporadically encountered among statutory provisions. Nevertheless, such requirement may be implicitly concluded from regulations for effective and high-quality service provision. Evidence-based rehabilitation medicine differs from evidence-based clinical medicine in that rehabilitation is obliged to aim at more therapeutic goals than just maintenance, recovery, or improvement of health. Furthermore, among statutory pension funds the granting of rehabilitation measures depends on an individual's probability to improve his or her earning capacity by rehabilitation. This point particularly denotes the need for a sound scientific evidence base. Care, as defined by Social Code Book XI (SGB XI), is mainly directed towards the support of daily living activities- therefore a scientific evidence base does not play a specific role. In sick nursing, the role of evidence-based practice has not yet achieved a clear significance. Aside from service provision, evidence-based decisions may play a role in planning of facilities and services for rehabilitation. In rehabilitation medicine, the responsibilities for the production and financing of scientific evidence, aside from educational and research purposes, may be seen in compatibility with the responsibilities after and 19, Para. 1 Social Code Book IX for the provision of adequate services: The federation, the states, and providers/payers of rehabilitation. PMID- 15804127 TI - [Indication guidelines for medical rehabilitation in the context of disease management programmes]. AB - In current and upcoming disease management programmes in Germany, the provision of medical services is strongly oriented on ICD diagnoses and on services traditionally provided by the statutory health insurance. Multidisciplinary services, such as medical rehabilitation, mostly covered by other payers (e.g. pension funds) are not taken into account. On the other hand, many chronically ill patients have complex and multifocal health complaints that are best addressed by multidisciplinary interventions. Considering this inherent deficit, in 2002 the German Society of Rehabilitation Sciences has initiated the research project "Indication Guidelines" aimed at developing indication criteria for rehabilitation in the context of disease management programmes. The concept presented in this paper relies on three basic requirements: 1. Impaired participation (according to ICF) caused by multifocal deficits leads to the definition of goals for rehabilitation, taking into account clinical and legal aspects as well as the patients preferences. 2. Multifocal health problems are best addressed by a multidisciplinary rehabilitation programme as it is currently provided by the German pension funds. 3. Scientific evidence has to demonstrate that these programmes are very likely to be effective (positive rehabilitation prognosis, evidence-based rehabilitation). Further requirements include adequate instruction of patients, as well as intensive and prolonged after-care. Both could be very well integrated into comprehensive disease management programmes. PMID- 15804128 TI - [Rehabilitation guidelines as tool of quality assurance in rehabilitation]. AB - The rehabilitation guideline program initiated by the German Federal Pension Insurance for Salaried Employees (BfA) can be regarded as the introduction of evidence- and consensus-based guidelines with direct relevance to rehabilitation practice in this specific sector of care. After successful implementation of the program, a substantial reduction of practice variation among rehabilitation institutions may be expected, hereby promoting the quality assurance in rehabilitation care. The program will also support the legitimization of medical rehabilitation as offered by the pension funds. So far, this is the first and only health care sector that has included the use of evidence-based practice guidelines routinely and ubiquitously into quality assurance activities. PMID- 15804129 TI - [The Rehabilitation Guideline Program of the BfA]. AB - Since 1998, the German Federal Pension Insurance for Salaried Employees (BfA) has funded several research projects aimed at developing clinical practice guidelines for medical rehabilitation. Using a standardized protocol, therapeutic processes for individual disorders were evaluated as to whether they were evidence-based. The projects covered about half of all rehabilitation cases handled by the BfA. Effective therapeutic procedures were identified in a systematic literature review. Clusters of therapies--so called evidence-based therapeutic modules (ETM) -were formed, outlining the frame of an ideal rehabilitation. A target/actual comparison was carried out to determine the need to develop rehabilitation guidelines. Routine data of the Classification of Therapeutic Performances derived from discharge letters were compared with the ETM, allowing comparison of the actual provision of treatment with the scientifically-defined target. The variance in therapies between the ETM on one hand, and among rehabilitation centers on the other hand, made the necessity for the development of rehabilitation clinical practice guidelines clear. The ETM were finalized after a written survey including all professions and a subsequent multi-professional expert workshop. The guidelines are now integrated into the BfA's quality assurance system. The process of implementation is accompanied by research teams. CONCLUSION: By developing clinical practice guidelines specific to rehabilitation, the pension insurance is the only sector of the German health system in which quality evaluation is carried out on the basis of clinical practice guidelines. This will improve the treatment of chronically ill patients on a scientific and quality-assured basis. PMID- 15804130 TI - [Can routine data on in-patient rehabilitation of musculoskeletal diseases replace the physicians' judgement in quality management? --an investigation on 12,014 observations]. AB - The acceptance of quality management is strongly influenced by the additional work-load necessary to obtain data. "Quality management questionnaire? Do we really need it? In rehabilitation, we must deliver a long and detailed discharge report anyway...". This frequent comment from the physician side has prompted us to investigate whether compilation of quality-management questionnaires really provides new information, or if a prediction model (for example similar to that predicting early retirement based on routine data in a discharge report) could be used to replace the physician judgement. If the prediction accuracy of the model is sufficiently high, the compilation of quality-management questionnaires might become obsolete, sparing the paperwork load of the physician. A central issue in the quality-management system of the LVA Baden-Wuerttemberg (a regional pension insurance institute) is the mutual agreement on a certain rehabilitation goal by patient and physician at the beginning of therapy. At the end of the rehabilitation program, the achievement of the goal is discussed with the patient in a discharge interview. In addition, patient and physician fill out a structured questionnaire with specification on three rehabilitation dimensions (WHO standard ICF: Body function and structure, activity, and participation). However, the results of multivariate modelling of 10 routinely observed parameters on more than 12,000 observations in patients with musculoskeletal diseases demonstrate that the agreement between the prediction model and the physician judgement of rehabilitation success is far away from optimal values. Thus, in this case the physician judgement can not be replaced by a forecast model solely based on routine data. Accordingly, the evaluation of rehabilitation success within the framework of a quality-management system based on currently valid standards must directly rely on the physician judgement. PMID- 15804131 TI - [Signals for the initiation of structured diagnostic procedures for depression in primary health care. A practice-relevant evaluation of international guidelines]. AB - BACKGROUND AND OBJECTIVES: Almost every tenth patient of a general practitioner (GP) suffers from depression. However, only 20-25% of these patients are correctly diagnosed during a GP consultation. How do international guidelines for depression in primary care initiate structured diagnostic procedures for depression? METHODS: We performed a systematic literature search on guidelines for the diagnosis of depression with focus on primary care. The quality of the guidelines was rated according to base of evidence, existence of pilot studies, data on implementation, presentation and specificity for primary care settings, and conflict of interest. We also screened whether and how the guidelines comment on the initiation of structured diagnostic procedures for depression. RESULTS: Of the 22 identified guidelines, only 15 address primary care. Only 3 of these were tested in pilot studies, 3 provided data on implementation, 9 were evidence based. The best guideline (6 out of 6 criteria met) is available in Dutch and established for The Netherlands only. We ranked the guidelines from NHG, VHA and ICSI as very good in terms of methodological quality. They present 'red flags' that initiate structured diagnostic procedures by 'opportunistic screening'. This is followed by the application of a self-rating instrument and an ICD-10-based diagnostic checklist identifying up to 98% of all patients with depression in a given consultation time of 10 minutes on average. CONCLUSION: Based on these criteria a national diagnostic depression guideline should, from our point of view, explicitly include keys such as "red flags" for the initiation of structured diagnostic procedures. PMID- 15804132 TI - [Promotion and value of education programs for patients with Type 1 and Type 2 diabetes mellitus, asthma and COPD, hypertension and breast cancer in Germany (Z Arztl Fortbild Qualitatssich. 2004 Aug;98(5):393-402)]. PMID- 15804133 TI - [Health technology assessment: stepchild of dentistry]. PMID- 15804134 TI - [Quality circle work in the field of the cash payment physicians' union of South Wurttenberg in 2003--numbers of contract physicians and contract psychotherapists and the continuing education supply and demand for moderators]. PMID- 15804135 TI - [Role of digital X-ray study in the current diagnosis of gastric cancer]. AB - The authors attempted to assess and provide evidence for the expedience of using a digital radiography CR system in clinical gastroenterology. The prerequisite for this was the results of large-scale diagnostic studies of different organ and systemic diseases. The authors underline the specific features of application of this digital system: the latter allows several analogue X-ray apparatuses to be transformed to digital ones, provides economic efficiency as compared with apparatuses with the direct digitization of an image, shows telemedical prospects, and has low radiation loads as evidenced by research and experimental studies even in comparison with green systems, which makes the use of these digital systems in X-ray gastroenterology highly tempting. Based on a great body of data from 126 studies, the authors could show the effectiveness of the digital radiography CR system in the diagnosis of gastric cancer, demonstrate a modernized approach to formatting an obtained digital image, the possibility of postprocessor treatment that can enhance the validity of existing X-ray symptoms. This all permitted the authors to recommend using the CR systems in the diagnosis of gastric cancer, by taking into account the current obvious predominance of its morphological diffuse and mixed forms and corresponding difficulties of their endoscopic diagnosis. PMID- 15804136 TI - [Standardizing a protocol of magnetic resonance imaging of temporomandibular joints. Part 2. Unification of analysis of obtained data]. AB - The paper presents a unified protocol for analyzing the data obtained by magnetic resonance tomography, which has been used to examine 350 patients. It characterizes the MR semiotics of different pathological conditions of articular structures, which are illustrated by MR images. An optimal terminology is proposed for the evaluation of bone and soft tissue changes. PMID- 15804137 TI - [A model of using magnetic resonance imaging in osteoarticular tumor lesion in case of giant cell tumors]. AB - Fifty-eight patients with giant cell tumors (GCT) underwent a comprehensive radiation diagnosis involving X-ray study and magnetic resonance imaging (MRI). The obtained MR images indicated the high efficiency of this combination of radiation diagnostic techniques in solving the problems in the visualization of osteoarticular tumor lesions. GCT is characterized by well-known primary X-ray semiotics; MR images are also rather pathognomonic of these tumors and they illustrate the process of morphogenesis of these masses. MRI made it possible to solve the specific problems facing a physician (a radiation diagnostician), to determine the site, shape, sizes, volume, and local extent of a tumor, which permitted the planning of surgical treatment policy; to assess its results, to reveal possible inflammatory complications; and to visualize a local recurrence and on-going growth of a tumor, including the signs of GCT malignancy. PMID- 15804138 TI - [Contrast-enhanced MRA of abdomen and lower limbs arteries: artifacts and pitfalls]. AB - The purpose of the work was the analysis of the most widespread diagnostic mistakes in connection with the technical artifacts arising at performing contrast-enhanced MR-angiography of abdomen and lower limbs arteries. PMID- 15804139 TI - [Three-dimensional visualization of low-field MRI data using personal computer]. AB - An experience of three-dimensional reconstruction of low-field MRI data using a modern personal computer and Windows software is presented. A value of volume visualization in routine diagnostics and areas of its effective use are shown. Pulse sequences best suitable for volume reconstruction are selected. Performance of high-end PC in visualization of medical data is evaluated. PMID- 15804140 TI - [Some problems in the creation of the telemedicine network of Russia and possible ways of their solution]. PMID- 15804141 TI - [Efficiency of use of gadolinium drugs as contrast substances]. PMID- 15804142 TI - Advances in atopic dermatitis. AB - Atopic dermatitis (AD) is a highly pruritic, chronic, and relapsing inflammatory skin disorder affecting 10-20% of children worldwide. During the past year there have been significant advances in our understanding of the cellular and immunologic mechanisms underlying AD as well as the immunologic triggers involved in its pathogenesis. The introduction of a new class of topical anti-inflammatory medications, topical calcineurin inhibitors, has significantly increased our treatment options and led to the rethinking of potential management approaches in AD. PMID- 15804143 TI - Rush pin fixation versus traction and casting for femoral fracture in children older than seven years. AB - BACKGROUND: The optimal treatment for femoral fractures in children is controversial. The purpose of this study was to compare the results of Rush pin fixation with those of conservative treatment, and to evaluate the sequels of growth plate injury by internal fixation. METHODS: Eighteen femoral shaft fractures in 17 children who had concomitant head injury or multiple traumas were treated surgically. The mean age at operation was 9 years 3 months (range, 7 years 5 months to 11 years 1 month). One Rush pin was inserted from the tip of the greater trochanter, without reaming, to fix the fracture. Another 20 age matched children treated by traction and casting were the control subjects. RESULTS: All the fractures united without consequences. In addition to a decrease in hospital stay with the use of the Rush pin (10 days vs. 27 days, p<0.05), fewer leg length discrepancies (4.2 mm vs. 7.1 mm, p<0.05) were also noted, compared with conservative treatment. The growth of the proximal femur after Rush pin fixation was evaluated after an average of 59 months. No femur shortening, coxa valgus, or hip dysplasia was noted. CONCLUSIONS: Intramedullary Rush pin fixation for femoral shaft fracture in children older than 7 years is a simple and reliable alternative. One narrow and non-reaming pin inserted from greater trochanter did not demonstrate femoral growth inhibition. PMID- 15804144 TI - Combination chemotherapy with carmustine and cisplatin followed by procarbazine, lomustine, and vincristine for adult high-grade astrocytoma. AB - BACKGROUND: We have reported that carmustine (BCNU) and cisplatin administered before, during, and after radiotherapy did not improve the survival of patients with high-grade astrocytomas and were associated with more serious toxicities than radiotherapy plus BCNU. In an attempt to improve survival, we studied a combination regimen procarbazine, lomustine, and vincristine (PCV) after radiotherapy in addition to BCNU and cisplatin during radiotherapy. METHODS: From 1994 through 1998, 42 patients were enrolled in the study. Of these, 20 had glioblastoma multiforme and 22 had anaplastic astrocytoma. The patients had a median age of 48.5 years. All patients had subtotal or total resection, or biopsy as the initial procedure. Then, all patients were treated with BCNU and cisplatin concurrently during radiotherapy followed by PCV after radiotherapy. RESULTS: The median time to follow up for survivors was 13.8 months (range, 1.7-108.2 months). The median time to tumor progression was 7.2 months (range, 0-88.7 months) and median survival time was 13.3 months (range, 1.7-88.7 months). The only factor that had a conventionally significant effect on the overall survival was resectability. Patients who had received subtotal/total resection had a longer median survival compared with patients who had received biopsy only (18.0 vs. 9.5 months). This combined modality treatment program was associated with reversible grade 3 to 4 hematological toxicity in 10 patients, with grade 3 ototoxicity in one patient and grade 2 neurotoxicity in one patient. CONCLUSION: A combination of BCNU and cisplatin with cranial irradiation followed by PCV was moderately toxic and appeared to offer no obvious survival advantages compared with radiotherapy plus BCNU and cisplatin alone. PMID- 15804145 TI - Diabetic ketoacidosis: comparisons of patient characteristics, clinical presentations and outcomes today and 20 years ago. AB - BACKGROUND: The aim of this study was to compare the clinical manifestations and outcomes of patients with diabetic ketoacidosis (DKA) today and 20 years ago. METHODS: A retrospective review was conducted of patients with DKA treated at our hospital from January 2001 through June 2002. The medical records were analyzed to identify clinical presentations, contributory factors, laboratory data, and outcomes. Additionally, data were compared with the records of patients with DKA in 1981 and 1982 at the same hospital. RESULTS: Data on 132 patients with 148 DKA episodes were included in the present study. When compared with the data from 20 years ago, clinical presentations, precipitating factors and laboratory data were similar. However, the mortality rate markedly decreased from 7.96% to 0.67%. Among patients with documented bacterial infections, a significantly high prevalence (70%) of Klebsiella pneumoniae infection was discovered. In patients with recurrent DKA, young women with type 1 diabetes accounted for most of the cases (67%) due to the omission of insulin. Eleven of 49 patients (22%) with newly diagnosed diabetes presenting with DKA were not insulin dependent during the 1.5-year follow up. CONCLUSIONS: After 20 years, the clinical presentations and precipitating factors of DKA were similar. However, the mortality rate was significantly reduced. Klebsiella pneumoniae was the leading cause of bacterial infections precipitating DKA in our hospital. Young women with type 1 diabetes were at high risk of repeat DKA. PMID- 15804146 TI - Incidence and risk factors of medical complications during inpatient stroke rehabilitation. AB - BACKGROUND: Knowing the types and frequency of medical complications of stroke patients during inpatient rehabilitation, together with the ability to identify high-risk patients, would be extremely helpful for providing the best possible physiologic environment for their recovery. METHODS: A retrospective study was conducted. Charts of 346 stroke patients who were consecutively admitted or transferred for inpatient rehabilitation in a tertiary care hospital from October 1997 to September 1998 were reviewed. Demographic and stroke characteristics, preexisting neuromedical conditions, neurological deficits, disability level, medical complications occurring during inpatient rehabilitation, and complications that required transfer off the rehabilitation ward were recorded for each patient. Univariate and stepwise multiple logistic regression analyses were performed to determine the factors that were significantly associated with the occurrence of any medical complication. RESULTS: Forty-four percent of patients experienced 1 or more complications. The most common complications were musculoskeletal pain (15.0%), urinary tract infection (13.6%), depression (9.3%), upper gastrointestinal tract bleeding (4.9%), and pneumonia (4.9%). Ten patients were transferred to the acute care ward during admission because of severe medical complications. Among them, 5 had a recurrent stroke, 3 had pneumonia, 1 had ventriculoperitoneal shunt obstruction, and 1 had organic psychosis. The occurrence of any of the medical complications was significantly associated with the following factors which were presented at admission or transfer to the rehabilitation ward: being female, having no voluntary movement of the affected hand, being more functionally dependent, and having to undergo Foley catheter insertion. CONCLUSIONS: Medical complications were common among stroke patients undergoing inpatient rehabilitation. Prevention and proper management are also very important in the rehabilitation ward. PMID- 15804147 TI - Retinopathy of prematurity: an evaluation in the Keelung area of Taiwan over a 10 year period. AB - BACKGROUND: We report on the experience of retinopathy of prematurity (ROP) at Chang Gang Memorial Hospital (CGMH), Keelung, Taiwan over a 10-year period. METHODS: A retrospective review was made of data of all premature infants with a diagnosis of ROP at Keelung CGMH between 1994 and 2003. Data on certain characteristics including gender, gestational age (GA), birth body weight (BBW), stage of ROP, and treatment modalities were collected and analyzed. RESULTS: Among the 458 infants screened for ROP, 148 eyes of 74 premature infants were diagnosed as having ROP. Threshold ROP occurred in 24 eyes of 12 infants. The average BBW and GA were significantly lower in the threshold than in the non threshold ROP group (p < 0.05). According to medical records of the 24 eyes of the 12 patients receiving transscleral cryotherapy, anatomical success was attained in 13 (81.3%) of 16 eyes. CONCLUSIONS: This review found low GA and BBW to be major risk factors for ROP. This review shows that transscleral cryotherapy for treating threshold ROP achieved anatomical success in about 80% of eyes. However, myopia, amblyopia, and strabismus remain major sequelae. Laser therapy is now believed to be less damaging to ocular structures and just as effective as cryotherapy in treating ROP. PMID- 15804148 TI - Perioperative hypertensive crisis in clinically silent pheochromocytomas: report of four cases. AB - Improvements in imaging have resulted in an increase in incidentally discovered adrenal tumors. The adrenal incidentalomas have been identified in at least 2%-3% of patients receiving abdominal computed tomography (CT), and pheochromocytomas are reported to occur in about 5.1%-23.0% of those patients. Only a few case reports of clinically silent pheochromocytomas have been published in the English literature. Herein we present 4 cases of pheochromocytomas as unforeseen adrenal masses on abdominal images. The characteristics of these tumors were that they (1) were noted to have no particular symptoms, (2) were confirmed biochemically and pathologically with immunophenotypic staining, (3) were benign in nature, and (4) produced apparent blood-pressure fluctuations during the operation. Three of these patients underwent a hypertensive crisis during surgery, and antihypertensives were applied transiently for blood pressure control. Two cases suffered from dramatic hypotension after the tumor had been excised, and intravenous fluid expanders and ephedrine were necessary to maintain adequate blood pressure. The above changes in blood pressure had no correlations with the size of the tumors. Although these tumors were clinically silent, they indeed were biologically active. Surgical resection of these tumors was indicated, and the risk of complications, such as hypertensive or hypotensive crises, may be no less than those of other typical tumors. Thus, such tumors should be managed with great caution. PMID- 15804149 TI - Concomitant bilateral orbital and brain abscesses--unusual complications of pediatric rhinosinusitis. AB - Rhinosinusitis is a common medical problem. Complications involving the orbits or brain are not frequently met. Concomitant involvement of both orbits and the brain is very rare. We here describe a 6-year-old girl who suffered from acute rhinosinusitis and initially, subperiosteal abscesses in both orbits. Bilateral functional endoscopic sinus surgery and drainage of the subperiosteal abscesses through the upper eyelids were done. Pus cultures showed methicillin-resistant Staphylococcus aureus. The brain abscesses were found under magnetic resonance imaging (MRI) and were treated conservatively with vancomycin and rifampin for 7 weeks. The patient was free of ouular and necurologic sequelae at the 1-year follow up. Retrograde thrombophlebitis instead of anatomic dehiscence was the most likely route for the spread of infection. Multidisciplinary approaches are necessary in managing ophthalmologic and intracranial complications of sinusitis. PMID- 15804150 TI - Hereditary neuropathy with liability to pressure palsies: a clinical and genetic study of a Taiwanese family. AB - Hereditary neuropathy with liability to pressure palsies (HNPP), an autosomal dominant disorder, is characterized by recurrent isolated nerve palsies, which are precipitated by trivial compression or trauma. In this report, we present the clinical features, electrophysiological studies, nerve biopsy results, and molecular analyses of a Taiwanese family. Among the 7 members evaluated, one latent and three symptomatic patients were found who showed a heterogeneous presentation from asymptomatic to characteristically recurrent peripheral neuropathy. Electrophysiological studies revealed a general decrease in nerve conduction velocities in all four patients with focal conduction slowing, especially at the compression sites. A sural nerve biopsy with a teased fiber preparation in the index patient demonstrated a typical tomaculous appearance. Molecular genetic studies exhibited a deletion of the PMP22 gene in chromosome 17p11.2-12 in all 4 patients. In conclusion, the diagnosis of HNPP might be overlooked if based on clinical presentation only. Family survey and electrophysiological and genetic tests should be done to investigate this disorder. PMID- 15804151 TI - Combination of acute preoperative plateletpheresis, cell salvage, and aprotinin minimizes blood loss and requirement during cardiac surgery. AB - Acute preoperative plateletpheresis (APP), cell salvage (CS) technique, and the use of aprotinin have been individually reported to be effective in reducing blood loss and blood component transfusion while improving hematological profiles in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB). In this prospective randomized clinical study, the efficacy of these combined approaches on reducing blood loss and transfusion requirements was evaluated. Seventy patients undergoing primary coronary artery bypass grafting (CABG) were randomly divided into four groups: a control group (group I, n = 10) did not receive any of the previously mentioned approaches. An APP and CS group (group II, n = 20) experienced APP in which preoperative platelet-rich plasma was collected and reinfused after reversal of heparin, along with the cell salvage technique throughout surgery. The third group (group III, n = 22) received aprotinin in which 5,000,000 KIU Trasylol was applied during surgery, and a combination group (group IV, n = 18) was treated with all three approaches, i.e., APP, CS, and aprotinin. Compared with group I (896+/-278 mL), the postoperative total blood loss was significantly reduced in groups II, III, and IV (468+/-136, 388+/-122, 202+/-81 mL, respectively, p < 0.05). The requirements of packed red blood cells in the three approached groups (153+/-63, 105+/-178, 0+/-0 mL, respectively) also were reduced when compared with group I (343+/-118 mL, p < 0.05). In group I, six patients (6/10) received fresh-frozen plasma and three patients (3/10) received platelet transfusion, whereas no patients in the other three groups required fresh-frozen plasma and platelet. In conclusion, both plateletpheresis concomitant with cell salvage and aprotinin contribute to the improvement of postoperative hemostasis, and the combination of these two approaches could minimize postoperative blood loss and requirement. PMID- 15804154 TI - Accuracy of temperature measurement in the cardiopulmonary bypass circuit. AB - Oxygenator arterial outlet blood temperature is routinely measured in the cardiopulmonary bypass (CPB) circuit as a surrogate for the temperature of the arterial blood delivered to sensitive organs such as the brain. The aim of this study was to evaluate the accuracy of the temperature thermistors used in the Terumo Capiox SX25 oxygenator and to compare the temperature measured at the outlet of the oxygenator using the Capiox CX*TL Luer Thermistor with temperatures measured at distal sites. Five experimental stages were performed in vitro to achieve this aim. Under our experimental conditions, the luer thermistors accurately measured the temperature as referenced by a precision thermometer. In the CPB circuit, the difference between arterial outlet and reference thermometer temperature varied with outlet temperature over-reading at low temperatures and under reading at high temperatures. There was negligible heat loss (-0.4+/ 0.1degrees C) measured at 4.5 m from the arterial outlet. The Terumo Capiox CX*TL Luer Thermistor is an accurate and reliable instrument for measuring temperature when incorporated into the Capiox Oxygenator. The accuracy in the measurement of temperature using these thermistors is affected by the thermistor immersion depth. Under reading of the arterial blood temperature by approximately 0.5 degrees C should be considered at normothermic temperatures, to avoid exceeding the maximum arterial blood temperature as described by institutional protocols. The accuracy of blood temperature measurements should be considered for all oxygenator arterial outlet temperature probes. PMID- 15804155 TI - Temperature inaccuracies during cardiopulmonary bypass. AB - Cerebral hyperthermia caused by perfusate temperature greater than 37 degrees C during the rewarming phase of CPB has been linked to postoperative neurologic deficits. The purpose of this study was to determine the accuracy of the coupled temperature measurement system and the CDI 500 arterial temperature sensor. Seventeen patients undergoing CPB were divided into four groups, each with a different temperature probe coupled to the oxygenator. The coupled temperature measurement system and CDI temperature sensors were compared with an indwelling probe placed in direct contact with the arterial perfusate. Blood, bladder, room and water temperatures, arterial line pressure, blood flow, and hemoglobin were recorded while the patients were supported with CPB. The actual blood temperature was significantly higher than the coupled temperature measurement system for two of the four groups (mean = 1.61 degrees C and 0.91 degrees C, p < 0.0001). A significant positive correlation between the actual temperature and the coupled temperature measurement system error was observed for the same two groups (r = 0.44, p < 0.0001). The actual temperature was significantly higher than the CDI temperature in all patients (mean = 1.2 degrees C, p < 0.0001). The coupling mechanism on the oxygenator generates inconsistent temperature readings. The perfusionist should consider these inconsistencies when using coupled temperature measurements and may consider the use of a direct temperature measurement system. The CDI temperature error is probably the result of inadequate flow through the sensor. On the test circuit, the flow of 170 mL/min was inadequate for circuit temperature accuracy. The accuracy of the CDI temperature drastically improved when the flow-through the sensor was increased to approximately 400 mL/min. Thus, the perfusionist must ensure adequate flow through the sensor in order for the temperature mechanism to function properly. Finally, the perfusionist can prevent cerebral hyperthermia by not allowing water temperature to exceed 37 degrees C, when using a coupled temperature measurement system. PMID- 15804153 TI - Clinical evaluation of poly(2-methoxyethylacrylate) in primary coronary artery bypass grafting. AB - In an attempt to make cardiopulmonary bypass (CPB) less traumatic for patients undergoing cardiac surgery, extracorporeal circuits (ECC) have been modified to achieve this goal. Poly(2-methoxyethylacrylate) (PMEA, X-coating) is a new polymer coating used in the ECC. PMEA studies have shown excellent biocompatibility with the components of blood. In this evaluation, PMEA-coated ECC were compared with control (CTR) circuits with emphasis on hematological parameters, peri-operative homologous blood product usage, and clinical outcomes. Patients undergoing elective coronary artery bypass grafting were randomized to either a PMEA group (n = 30) or a CTR group (n = 30). Extracorporeal circuit components in the PMEA group were coated except for the cardioplegia delivery device and cannulas. Patients in the CTR group had just the arterial line filter coated. The following hematological parameters were measured: platelet count (PLT), white blood cell count (WBC), red blood cell count (RBC), and hematocrit (Hct). Blood product usage was observed along with clinical outcomes for the following parameters: ventilation time, mediastinal tube output, intensive care unit (ICU) and hospital lengths of stay. The preoperative patient profiles were comparable between the two groups. The PMEA group had marginally higher CPB times (134+/-31.9 vs. 118+/-33.7 minutes) and cross clamp times (83.9+/-21.3 vs. 73.7+/ 21.6 minutes), however no significant differences were reached. Platelet count, RBC, and Hct levels were also comparable between groups with no significant differences. However, there was a significant difference in WBC between groups (p = 0.041). Less platelets were administered both intraoperatively and 48 hours postoperatively in the PMEA group. The authors evaluated PMEA-coating by measuring clinical outcomes, such as ventilation time, ICU and hospital lengths of stay, and homologous blood utilization. PMEA patients trended towards less homologous blood transfusions, which helped save an average of $83.41 per patient. Further clinical studies are needed to evaluate the benefits of this new polymer coating. PMID- 15804157 TI - Quantifying platelet gel coagulation using Sonoclot and Thrombelastograph hemostasis analyzer. AB - Little in vitro research exists discussing platelet gel composition and the resulting strength and degradation characteristics using point-of-care technologies. There must be a quantifiable way of determining the structural integrity of the resulting formed platelet gel thrombus. The Thrombelastograph Hemostasis Analyzer (TEG) and Sonoclot measure the elasticity of a clot as it forms and subsequently degrades naturally. The objective of this study was to determine the application of TEG and Sonoclot technologies as point-of-care devices for technicians using platelet gel therapy. The collected bovine blood was anticoagulated with CPD and processed using a previously published plasma sequestration protocol, using normal saline as a wash solution. The resulting platelet-rich plasma was stored in a sequestration bag in a water bath to maintain the blood temperature at 37 degrees C. Sequestered bovine platelet-rich plasma was made into platelet gel using three different thrombin concentrations. A total of 30 experiments were performed on the platelet gel product using both the TEG and the Sonoclot. We discovered that 6 of the Sonoclot tests and 15 of the TEG tests were valid. None of the TEG clot signatures and nine of the Sonoclot signatures were discovered to be invalid. A chi2 test was performed on the resultant data. The value of the chi2 test was calculated to be 12.86, which translated into a p value of less than 0.001. Despite the vast use and growing popularity of platelet gels, a method in which to quantify platelet gels has yet to be reported. There remains a possibility that gels formed with different concentrations of components may prove useful in different areas of surgery or their uses may expand to a broader spectrum of medicine. However, technology to quantify platelet gels must first be standardized. On the basis of the data collected in this study, it was determined that the TEG and the Sonoclot are not equally capable of analyzing platelet gel clots. The TEG is a valid means for analysis, whereas the Sonoclot provided unreliable analysis based on a Chi squared test. PMID- 15804156 TI - Is platelet function as measured by Thrombelastograph monitoring in whole blood affected by platelet inhibitors? AB - Platelet inhibitors, especially the glycoprotein (GP) IIb/IIIa receptor antagonists, have demonstrated their effectiveness in reducing the acute ischemic complications of percutaneous coronary intervention (PCI) and in improving clinical outcomes in patients with acute coronary crisis. Three common platelet inhibitors observed in emergent cardiopulmonary bypass (CPB) for failed PCI are abciximab, eptifibatide, and tirofiban. An in vitro model was constructed in two parts to determine whether platelet aggregation inhibition induced by platelet inhibitors would be demonstrated by the Thrombelastograph (TEG) monitor when compared with baseline samples with no platelet inhibitor. In part A, 20 mL of fresh whole blood was divided into four groups: group I = baseline, group A = abcix-imab microg/mL, group E = eptifibatide ng/mL, and group T = tirofiban ng/mL. Platelet inhibitor concentrations in whole blood were derived starting with reported serum concentrations with escalation to achieve 80% platelet inhibition using the Medtronic hemoSTATUS and/or Lumi-aggregometer. A concentration range determined by our in vitro tests were chosen for each drug using concentrations achieving less than, equal to, or greater than 80% platelet inhibition. In part B, TEG analysis was then performed using baseline and concentrations for each drug derived in part A. Parameters measured were clot formation reaction time (R), coagulation time (K), maximum amplitude (MA) and alpha angle (A). Groups E1000 and E2000 extended R over control by 37% and 23%, respectively (p = 0.01 and 0.03). Groups E1000 and E2000 increased K times by 45% and 58% (p = .02 and .04). T160 samples prolonged K by 20% (p = 0.01). The angle or clot strength (A) was decreased in groups T160 and E1000 by 23% (+ 7.06 SD) and 18% (+ 11.23 SD), respectively (p = 0.001 and 0.01). The MA decrease was statistically significant in the T160, E1000 and E2000 by 9%, 6% and 13% respectively (p = 0.01). Samples treated with abciximab were comparable to control values for all parameters measured. Although statistical significance could be demonstrated with some parameters, sensitivity was only observed at increased doses and was not seen with all agents tested. In our in vitro model, the TEG monitor was unable to demonstrate clinically significant differences in platelet function and may not be reflective of platelet function in samples which have been treated with these GP IIb/IIIa inhibitors. PMID- 15804152 TI - Cardiopulmonary bypass in patients with pre-existing coagulopathy. AB - Patients with pre-existing coagulopathies who undergo surgical interventions are at increased risk for bleeding complications. This risk is especially true in cardiac surgical procedures with cardiopulmonary bypass (CPB) because of the necessity for heparinization and the use of the extracorporeal circuits, which have destructive effects on most of the blood components. In this review, cases of cardiac surgeries in patients with certain pre-existing coagulopathies are summarized, which could shed a light on future managements of such patients undergoing cardiac procedures with CPB. Pre-existing coagulopathies include antithrombin III deficiency, heparin-induced thrombocytopenia, cancer, factor XII deficiency, hemophilia, idiopathic thrombocytopenic purpura, protein S deficiency, and drug-induced platelet inhibition. In summary, pre-existing coagulopathy in patients undergoing open-heart surgeries, if not recognized and appropriately managed, can cause serious complications. Management of patients undergoing cardiac procedures should include a routine coagulation work-up and a thorough past medical history examination. If any of the foregoing is abnormal, further evaluation is warranted. Proper diagnosis and management of the pre existing coagulopathy disorders is of crucial importance to the surgical outcome and long-term morbidity. PMID- 15804158 TI - Thrombelastograph (TEG) analysis of platelet gel formed with different thrombin concentrations. AB - Autologous blood transfusion is the safest and most successful way to decrease transfusion-related risks such as postoperative infections, allo-immunization, and short- and long-term immunosuppression. In addition, these fibrin sealants are known to provide coagulation support at the surgical site and act as an adjunct to the control of postoperative bleeding. The physical formation of autologous platelet fibrin gel clot is dependent on both the common pathway of the coagulation cascade and platelet activation. Platelet gel can help provide control of intraoperative and postoperative bleeding. The Thrombelastograph Hemostasis Analyzer (TEG) measures the viscoelastic properties of a clot as it forms. Based on the information that the TEG provides, it promises to be a good choice for point of care measurement of the integrity of thrombus formed by platelet gels. Bovine blood from a single donor was sequestered into platelet rich plasma and was made into platelet gel using calcium and three different concentrations of thrombin. The platelet gel samples were then analyzed with the TEG analyzer. The results for MA, tMA, CI, and angle were recorded and statistical analysis was performed to accept or reject the null hypothesis, which is: There is no difference between TEG parameters when analyzing platelet gels formed with calcium chloride, platelet-rich plasma and three different concentrations of thrombin A one-way analysis of variance test was performed between thrombin concentrations for MA (p = 0.19), tMA (p = 0.443), CI (p = 0.257), and angle (p = 0.323). The results showed that thrombin concentration did not affect the MA, tMA, CI, or angle as measured by the TEG analyzer. The null hypothesis was accepted. Based on a one-way analysis of variance test for MA, tMA, CI, and angle there was no significant statistical difference for the TEG samples in this experiment as reported with a 95% confidence interval. PMID- 15804159 TI - Quality of red blood cells using the Dideco Electa autotransfusion device. AB - The purpose of this study was to evaluate the quality of washed, concentrated red blood cells (RBCs) produced by the new Electa autotransfusion device from Cobe Cardiovascular (Dideco). Blood was collected intraoperatively in 16 patients undergoing cardiac surgery for whom routine cell savage was being used and then washed using the Electa. According to the manufacture's protocol. 125-mL bowls were used in the standard wash program. Reservoir and washed RBCs were analyzed for platelets (PLTs), leukocytes (WBCs), potassium (K+), and plasma-free hemoglobin (PFH) removal, as well as, hematocrit (Hct) and RBC recovery. The Electa cell saver produced a product with an average Hct of 58+/-5% and a RBC recovery rate of 87+/-10%. Its removal of waste products resulted in the washout of 54+/-18% WBCs, 87+/-6% PLTs, 91+/-4% K+, and 77+/-17% PFH. The Electa produces a good-quality washed RBC product that is comparable with other autotransfusion devices on the market. PMID- 15804160 TI - Rat cardiopulmonary bypass model: application of a miniature extracorporeal circuit composed of asanguinous prime. AB - A clinically relevant rat cardiopulmonary bypass (CPB) model would be a valuable tool for investigating pathophysiological and therapeutic strategies on bypass. Previous rat CPB models have been described in the literature; however, they have many limitations, including large circuit surface area, the inability to achieve full bypass, and donor blood requirements for prime. Therefore, we have established a rat CPB model designed to overcome these limitations. The miniature circuit consisted of a filtered reservoir, heat exchanger, membrane oxygenator (surface area = 0.02 m2) with a static priming volume of 2.8 mL, and an inline blood gas monitor. The circuit was primed with 9.5+/-0.5 mL of crystalloid solution and CPB was established on male Sprague-Dawley rats (430-475 g, n = 5) by cannulating the left common carotid artery and the right external jugular vein. The animals were placed on CPB at full flow (111+/-13 mL/kg/ min) for 1 hour and were monitored for and additional 2 hours after the CPB procedure. Hemodynamics, hemoglobin concentration (Hb), and blood gases were analyzed at three time intervals: before, during, and after CPB. The circuit performance was evaluated according to prime volume, compliance, hemodynamic parameters, and gas and heat exchange as described by modified AMMI standards. Data are expressed as mean+/-SD and a repeated-measures analysis of variance with post-Hoc test was used for data comparison between the three time intervals. The ratio of oxygenator surface area to subject body weight for this model is comparable with that of current human adult CPB practice (0.05 m2/kg vs 0.057 m2/kg) Full CPB was achieved and we observed clinically acceptable PaO2, PaCO2, and SvO2 values (209+/-86 mmHg, 25+/-2 mmHg, 78+/-8%, respectively) while on CPB. The use of asanguinous prime did produce statistically significant Hg reduction (15.7+/-0.76 vs. 9.2+/-0.59 g/dL) comparable with clinical practice. No statistically significant differences between pre- and post-CPB hemodynamics and blood gases were found in our study. We have established a miniature circuit consisting of asanquineous prime for a rat CPB model that maintains clinically acceptable results regarding hemodynamic parameters, blood gases, and hemodilution. This model would be valuable for further use in clinically relevant research studies. PMID- 15804161 TI - Optimizing management of hirudin anticoagulation. AB - Accurate anticoagulation monitoring, critical during cardiac surgery (CS), is especially important for novel therapeutics such as hirudins, for which there are no known antidotes. The activated clotting time (ACT), which is standard for heparin monitoring, has been reported to be insufficiently sensitive to high levels of hirudins. A simple, accurate, and sensitive assay is needed to monitor hirudins at the levels required for CS. During the REPLACE/II clinical trials, the HEMOCHRON Jr. Signature ACT+ was used to monitor Angiomax during percutaneous cardiac intervention (PCI) procedures and was observed to lose sensitivity at bivalirudin concentrations greater than 8-10 microg/mL. A new assay, the ACTT, was developed to increase the linear sensitivity of the ACT+ over the range of 15 30 microg/mL bivalirudin to extend the clinical utility of the assay to CS levels. Both in vitro and ex vivo studies were performed using the ACTT and ACT+. In vitro ACT+ and ACTT clotting times, identical for bivalirudin levels up to approximately 5 microg/mL, diverged from each other near 10 microg/mL. The ACTT showed excellent linearity to bivalirudin at concentrations up to 30 microg/ mL. Reproducibility was also superior with coefficients of variation <15% across 13 donors at clotting times <760 seconds. The ACTT was evaluated for monitoring bivalirudin during PCI in 67 patients. The ex vivo comparison of ACTT to ACT+ <340 seconds, showed a slope near 1.0 and an average difference between the tests of 5%. At higher clotting times this slope increased to near 3.0, with an average difference between tests of 20%. These data suggest that the ACTT displays increased sensitivity to high levels of bivalirudin. PMID- 15804162 TI - Extracorporeal membrane oxygenation and CO2 removal in an adult after near drowning. AB - Near drowning is a common event among otherwise healthy young people. The development of ARDS in the setting may significantly increase mortality. The traditional means of ventilation may lead to barotrauma. Extracorporeal membrane oxygenation (ECMO) is an effective means to improve oxygenation and remove carbon dioxide, while allowing the lungs to recover from the acute insult. It may be especially successful in those victims with single organ injury. We report the use of ECMO in a young adult with ARDS and pneumonia after near drowning. PMID- 15804163 TI - Hemicolectomy with concomitant heated intraperitoneal chemotherapy: a case study. AB - The use of heated intraperitoneal chemotherapy is an emerging new adjunct in the treatment of adenocarcinoma of the colon. However, documentation regarding perfusion circuitry and techniques associated with this therapy remain largely undescribed. After consultation with the surgical service team, a custom designed circuit was constructed for this procedure. Institutional approval and informed consent were obtained for surgical debulking and heated intraperitoneal chemotherapy for a 58-year old female. After surgical resection, a right hemicolectomy was performed and pathological specimens obtained. A modified custom circuit using a roller pump was first primed with 3 liters of Dianeal PD-2 and recirculated until temperature of 41 degrees C was obtained. The circuit was then connected to the patient for infusion of perfusate via Blake drains placed in the deep pelvis. Two additional drains were placed in the subdiaphragmatic space for return. Perfusate containing 30 mg of Mitomycin-C was circulated at 600 800 mL/min for 60 min at 41 degrees C. An additional 10 mg of Mitomycin-C was then administered through the circuit for an additional hour under similar conditions. Upon completion, a washout procedure was performed with 2 additional liters of Dianeal PD-2. The patient tolerated the procedure well and was discharged postoperative day 7. We describe the successful use of a perfusion administered heated intraperitoneal chemotherapy regimen as an integral part of successful treatment of adenocarcinoma of the colon. PMID- 15804164 TI - The impact of the 2003 heat wave on mortality and hospital admissions in England. AB - This article quantifies the impact of the heat wave, 4 to 13 August 2003, on mortality and emergency hospital admissions in England by region and age group. The August 2003 heat wave was associated with a large short-term increase in mortality, particularly in London. Overall in England there were 2,091 (17 per cent) excess deaths. Worst affected were those over the age of 75. The greatest increase of any region in England was in London in the over 75 age group with 522 excess deaths (59 per cent). Excess hospital admissions of 16 per cent were recorded in London for the over 75s. Temperatures in England were unusually hot. Ozone and particulate matter concentrations were also elevated during the heat wave. Estimated excess mortality was greater than for other recent heat waves in the UK. PMID- 15804165 TI - Is the health of the long-term unemployed better or worse in high unemployment areas? AB - Data on 25.6 million adults from the UK 2001 Census were analysed to compare the regional pattern of self-rated health of the long-term unemployed to that of people from different social classes and of those who have never worked. The results show that the health of the long-term unemployed was better in high unemployment regions, and conversely, worse where the local labour market was traditionally stronger. This is the reverse of the regional pattern found-for different social classes and for those who have never worked. PMID- 15804166 TI - Healthy life expectancy by area deprivation: magnitude and trends in England, 1994-1999. AB - This article examines the magnitude of inequalities in health by area deprivation using two composite indices of health expectancy, one based on a subjective assessment of general health status (healthy life expectancy) and the second on reported limiting longstanding illness (disability-free life expectancy). Trends in healthy life expectancy by deprivation for the period 1994-1999 were also examined. Results show that males and females living in the most deprived wards spend twice as many years in poor health, both in absolute (years of life) and relative (proportion of life) terms, than those living in the least deprived wards. There was no change in the healthy life expectancy gap between the most and least deprived areas over the study period. PMID- 15804167 TI - Report: deaths related to drug poisoning: England and Wales, 1999-2003. PMID- 15804168 TI - Report: deaths involving MRSA: England and Wales, 1999-2003. PMID- 15804169 TI - Annual update: congenital anomaly statistics: notifications, 2003, England and Wales. PMID- 15804170 TI - Annual update: 2003 mortality statistics: cause (England and Wales). PMID- 15804172 TI - Local prevention of oxidative stress in the intestinal epithelium of the rat by adhesive liposomes of superoxide dismutase and tempamine. AB - The purpose of the present study was to investigate whether the local prevention of luminal superoxide-mediated biological damage in the rat jejunal mucosa could be achieved by liposomal superoxide dismutase (SOD) and the SOD mimic tempamine (TMN). Cationic liposomes loaded with either SOD or TMN were perfused in the rat jejunum prior to the induction of oxidative insult. Reactive hydroxyl radicals were generated in situ in a closed circulating intestinal loop of the rat from the reaction between hypoxanthine and xanthine oxidase in the presence of chelated ferrous sulfate. Mucosal activity of lactate dehydrogenase and levels of potassium ions were used to quantify the tissue damage. Intracellular uptake and locality of SOD were examined in HT-29 cells. The intestinal uptake of SOD and TMN was further measured by using rat colon sacs. Entrapment in cationic liposomes was found to significantly enhance the antioxidant effect of SOD and TMN against the induced oxidative damage in the jejunal mucosa, compared with their free forms. The effect was found to be local and was caused by the increased mucosal adhesion of the liposomes. The cationic liposomes also triggered SOD uptake into the HT-29 cell line. It is concluded that the increased residence time of the cationic liposomes of SOD and TMN in the jejunal mucosa resulted in a local effect against oxidative injury. This local protection may be exploited for drug delivery purposes. PMID- 15804173 TI - Functional role of P-glycoprotein in limiting intestinal absorption of drugs: contribution of passive permeability to P-glycoprotein mediated efflux transport. AB - The aim of the present study is to evaluate the quantitative contribution of passive permeability to P-glycoprotein-mediated (P-gp-mediated) efflux and the functional activity of P-gp in determining intestinal absorption of drugs, and demonstrate the relationship between efflux parameters and intestinal permeability. MDRI-MDCKII cell monolayer permeability, human intestinal absorption (HIA), and solubility data were systematically collected from the literature. Drugs were classified as a total of 63 P-gp substrates (P-gpS) and 73 nonsubstrates (NS) on the basis of efflux ratio or calcein AM inhibition and ATPase activity assays. Efflux parameters, efflux ratio (ER) and absorption quotient (AQ), were correlated to the monolayer permeability. MDRI-MDCKII cell monolayer permeability characteristics were found to be distinctly different between P-gpS and NS datasets. The ER for P-gpS was found to increase with absorptive permeability until 20 nm.s(-1), but reduced for P-gpS with high absorptive permeability. The AQ showed a linear inverse relationship with absorptive permeability. Overall, efflux parameters, ER and AQ, indicated that the transport of P-gpS with moderate passive permeability is highly attenuated by P-gp, while passive permeability overrules the P-gp-mediated efflux for high permeability molecules. Most of the P-gpS were found towards the upper limits of molecular weight (>500) and calculated total polar surface area (>75 A(2)). This dataset indicated that unfavorable chemical features of P-gpS limit passive permeability and thus are more susceptible to P-gp-mediated efflux. In conclusion, passive permeability versus P-gp-mediated efflux determines intestinal permeability of P-gpS, where P-gp limits absorption of only moderately permeable compounds. Thus, integrating these factors with drug characteristics of the Biopharmaceutics Classification System (BCS) class better predicts the functional role of P-gp in limiting intestinal drug absorption. PMID- 15804174 TI - Immunostimulation mechanism of LPD nanoparticle as a vaccine carrier. AB - A novel and improved vaccine delivery system and/or adjuvant is actively sought to enhance the potency of vaccines. Previously, we reported that strong antitumor immunity could be generated when a peptide antigen was incorporated into LPD (cationic liposome-polycation-pDNA) nanoparticles. In this study, we found that both the cationic liposome and DNA are required for the full immunostimulation activity of LPD. The unique ability of LPD to readily move into local lymphoid tissues and to activate antigen-presenting cells might be responsible for its strong immunostimulatory activity. Moreover, cationic liposome stimulates the expression of CD80/CD86 on dendritic cells (DCs), but not the release of TNF alpha from DCs, suggesting the existence of a NF-kappaB-independent immunostimulation pathway for cationic lipids such as DOTAP. PMID- 15804175 TI - Superior cell delivery features of poly(ethylene glycol) incorporated alginate, chitosan, and poly-L-lysine microcapsules. AB - Microencapsulation is an emerging technology in the development of bioartificial organs for drug, protein, and delivery systems. One of the advancements in establishing an appropriate membrane material for live cell and tissue encapsulation is the incorporation of poly(ethylene glycol) (PEG) to the widely studied alginate microcapsules. The current study investigates the properties of integrating PEG to microcapsules coated with poly-L-lysine (PLL) and chitosan as well as a novel microcapsule membrane which combines both PLL and chitosan. Results show that microcapsules containing PEG can support cell viability and protein secretion. The addition of PEG to PLL and chitosan-coated microcapsules improves the stability of microcapsules when exposed to a hypotonic solution. We also compared the novel microcapsule with two other previously used microcapsules including alginate-chitosan-PEG and alginate-PLL-PEG-alginate. Results show that all three membranes are capable of providing immunoprotection to the cells and have the potential for long-term storage at -80 degrees C. The novel membrane containing PEG, chitosan, and PLL, however, revealed the highest cell viability and mechanical strength when exposed to external rotational force, but it was unable to sustain osmotic pressure. The study revealed the potential of using PEG incorporated alginate, chitosan, and PLL microcapsules for encapsulating live cells producing proteins and hormones for therapy. PMID- 15804176 TI - Prolidase, a potential enzyme target for melanoma: design of proline-containing dipeptide-like prodrugs. AB - Bioinformatics tools such as Perl, Visual Basic, Cluster, and TreeView were used to analyze public gene expression databases in order to identify potential enzyme targets for prodrug strategies. The analyses indicated that prolidase might be a desirable enzyme target based on its differential expression in melanoma cancer cell lines and its high substrate specificity for dipeptides containing proline at the carboxy terminus. RT-PCR expression of prolidase and hydrolytic activity against N-glycyl-l-proline (GLY-PRO), a standard substrate of prolidase, determined in tumor cell lines, exhibited a high correlation (r(2) = 0.95). These results suggest the possibility of targeting prolidase with prodrugs of anticancer agents for enhanced selectivity. The feasibility of such a scenario was tested by (a) synthesizing prodrugs of melphalan that comprised linkage of the carboxy terminus of the l-phenylalanine moiety of melphalan to the N-terminus of l and d stereoisomers of proline and (b) determining their bioconversion and antiproliferative activities in SK-MEL-5 cells, a melanoma cancer cell line with high expression levels of prolidase. The results of hydrolysis studies of the l- and d-proline prodrugs of melphalan, designated as prophalan-l and prophalan-d, respectively, indicated a approximately 7-fold higher rate of activation of prophalan-l compared to prophalan-d in SK-MEL-5 cell homogenates. Prophalan-l exhibited cytotoxicity (GI(50) = 74.8 microM) comparable to that of melphalan (GI(50) = 57.0 microM) in SK-MEL-5 cells while prophalan-d was ineffective, suggesting that prolidase-specific activation to the parent drug may be essential for cytotoxic action. Thus, melphalan prodrugs such as prophalan-l that are cleavable by prolidase offer the potential for enhanced selectivity by facilitating cytotoxic activity only in cells overexpressing prolidase. PMID- 15804177 TI - A systemic route for drug loading to lymphatic phagocytes. AB - Lymph nodes are primary germination and proliferation sites for many types of pathogens. Maintaining therapeutic levels of appropriate chemotherapeutic agents in the lymph node tissue is critical for the treatment of both infection and cancer. This study was intended to develop a systemic route for loading lymph node phagocytes with drugs, using a lymph node specific nanocarrier. The latter is assembled as a 10-15 nm particle with a drug-carrying core and a phagocyte homing poly(1-->6)-alpha-d-glucose based interface. Biokinetics and microdistribution of the model carrier were investigated in vivo. Nanocarrier accumulation in lymph nodes reached 30-35% dose/g in central lymph nodes, with deposition in various phagocytic cell populations. The latter included cells harboring inhaled microparticles translocated to lymph nodes from the lungs. In view of the nanocarrier ability to transport and release significant amounts of various drug substances, the data suggests feasibility of systemic drug loading to lymphatic phagocytes and, through drug release, to the neighboring cells. PMID- 15804178 TI - Transcutaneous delivery of levodopa: enhancement by fatty acid synthesis inhibition. AB - The present investigation aimed at evaluating the role of fatty acid synthesis inhibition in enhancing transcutaneous delivery of levodopa (LD). Rat epidermis was treated with ethanol and various doses of cerulenin (an inhibitor of fatty acid synthase enzyme system) for reducing the normal level of fatty acids. Calcium chloride (0.1 mM) and/or verapamil (1 microM) were coapplied to cerulenin treated skin in order to modulate duration of epidermal perturbation. These treated skin portions were used for estimation of altered triglyceride content (an indicator of fatty acid synthesis), differential scanning calorimetry (DSC) analysis, and in vitro permeation of LD. Plasma concentration of LD was monitored in rats following topical application of various transdermal formulations. Application of cerulenin (0.1 or 0.15 mM/7 cm(2)) to viable rat skin inhibited approximately 60% triglyceride synthesis with respect to control at 2 h. Coapplication of calcium chloride (0.1 mM) significantly increased this inhibition, whereas verapamil application reduced this effect. The decrease in triglyceride content reduced the enthalpy of the lipid endothermic transition. The in vitro permeation of LD was enhanced 3-fold across skin excised after treatment with cerulenin. LD did not permeate across normal skin. The effective plasma concentration (C(eff)) of LD was achieved within 3 h and maintained till 10 h by a single topical application of a carbidopa-levodopa combination (1:4) to ethanol-perturbed cerulenin-treated skin. Coapplication of calcium chloride reduced the time lag to achieve C(eff) to 2 h and maintained it till 24 h. A single transdermal LD (64 mg) patch formulated with calcium chloride (0.1 mM) and cerulenin (0.1 mM) dissolved in a propylene glycol:ethanol (7:3) mixture seems to offer a noninvasive approach for transcutaneous delivery of levodopa. PMID- 15804179 TI - Selective downregulation of the MDR1 gene product in Caco-2 cells by stable transfection to prove its relevance in secretory drug transport. AB - Considerable interest is focused on overcoming multidrug resistance (MDR) in cancer chemotherapy. The in vitro experiments to characterize P-glycoprotein's (P gp) function and to decrease its effects have led to a variety of strategies such as addition of competitors or supplementation of the medium with oligonucleotides complementary to the 5'-end of the MDR1-mRNA. For the Caco-2 cell line, an in vitro model for absorption screening, expressing multiple transporters including P-gp, which pumps substances back into the apical solution, P-gp activity might mask other relevant transport proteins' activity. The objective of the present study was to construct a Caco-2 subline with reduced P-gp expression level. Caco 2 cells were transfected by electroporation with two different mammalian expression vectors, and the obtained subclones were investigated at RNA (Northern blotting, RT-PCR), protein (FACS analysis), and functional (transport studies) levels for reduction in P-gp expression. Northern blotting showed that the levels of transcription of the inserted gene were different among the several clones, but those results did not completely correlate with the FACS analysis for P-gp expression. The clones with the strongest reduction in P-gp expression detected by the FACS analysis also showed the lowest secretory fluxes of the P-gp substrate talinolol in transport studies. Repetition of FACS analysis after 7 and 24 months on 20 to 30 passage older subclones still showed reduction in P-gp expression and indicated that they are stably transfected. The new cell lines constructed in the present study provide the possibility to perform in vitro absorption studies in a cell system composed of differentiated enterocytes growing as a monolayer like the normal Caco-2 cell line but with a lower down to almost lacking expression of P-gp. PMID- 15804180 TI - Differential cytosolic delivery and presentation of antigen by listeriolysin O liposomes to macrophages and dendritic cells. AB - Delivery of antigenic protein to the cytosol of antigen-presenting cells (APCs), such as macrophages (MPhi) and dendritic cells (DCs), is required for an efficient CD8 T-cell-mediated immune response. We have previously shown that co encapsulation of antigenic protein inside pH-sensitive liposomes with listeriolysin O (LLO), a pore-forming protein of Listeria monocytogenes, generates efficient major histocompatibility complex class I (MHC I)-restricted immune responses both in vitro and in vivo. In this study, we sought to analyze the relative efficiency of LLO-mediated cytosolic delivery of liposomal antigen in two important APCs, macrophages and dendritic cells, by examining the sequential steps involved in antigen presentation to T-cells in cultured mouse bone marrow-derived MPhis (BMMPhis) and DCs (BMDCs). BMMPhis overall presented liposomal antigen better than BMDCs at a given concentration of liposomal antigen incubated with cells, and the trend was also observed after the presentation was normalized by the uptake of antigen. When soluble antigen was directly introduced into the cytosol, however, BMDCs presented the antigen more efficiently than BMMPhis. In addition, when the APCs were externally loaded with the antigenic peptide of the protein, BMDCs displayed a higher level of cell surface MHC I peptide complexes and presented the peptide more efficiently than BMMPhis. These results combined together suggest that LLO-mediated release of liposomal antigen from the endosomal/lysosomal compartment may be more pronounced in BMMPhis than in BMDCs, and further implicates differential activity of LLO and varying efficiency of LLO-mediated endosomal escape in different antigen-presenting cell types. PMID- 15804181 TI - Directed antigen presentation using polymeric microparticulate carriers degradable at lysosomal pH for controlled immune responses. AB - The types of the immune responses generated against an antigen are determined by the intracellular fate of the antigen. Endogenous antigens are processed in the cytoplasm and initiate cytotoxic T lymphocyte (CTL) activation. In contrast, exogenous antigens are degraded in the lysosome (or phagolysosome) of antigen presenting cells (APCs), and induce antibody-mediated immune responses and assist CTL activation. Therefore, maximizing a desired response by controlling delivery pathways is indispensable in vaccine development and immunotherapy. New cleavable microparticles have been prepared for use as protein-based vaccine carriers by polymerizing water soluble monomers including a newly developed aliphatic monomer with a pendant primary amine group and a cleavable acetal linkage with a wholly aliphatic cleavable acetal cross-linker. Incorporation of the cleavable amine monomer in the polymerization mixture increased the encapsulation efficiency of a model antigen, ovalbumin. Ex vivo assays showed that the composition of the particles greatly affected the magnitude and the pathway of antigen presentations, which determine the type of immune responses. The degradable particles synthesized with the new cross-linker enhanced MHC I antigen presentation 2-3-fold over nondegradable particles. It was also found that, by adding 10% cationic cleavable monomers to the microparticles, MHC I restricted antigen presentation was enhanced ca. 75 times over that achieved with nondegradable particles. The microparticles introduced in this study can be further used for targeting and gene delivery due to functionalizable and cleavable cationic monomers in addition to degradability. PMID- 15804182 TI - A branched fluorescent peptide probe for imaging of activated platelets. AB - Novel fluorescent probes for thrombi and activated-platelet detection were developed that were based on the glycoprotein IIb/IIIa (GP-IIb/IIIa) binding sequence, Pro-Ser-Pro-Gly-Asp-Trp. Linear, Pro-Ser-Pro-Gly-Asp-Trp-Aha-Gly Cys(Cy5.5)-NH(2) (1PF), and branched, (Pro-Ser-Pro-Gly-Asp-Trp-Aha)(2)-Lys-Gly Cys(Cy5.5)-NH(2) (2PF), fluorescent-labeled peptide probes were synthesized. A third probe, also branched, (Pro-Ser-Pro-Gly-Glu-Trp-Aha)(2)-Lys-Gly-Cys(Cy5.5) NH(2) (2CF), was synthesized as control. The platelet-binding activity of the probes was tested in clots generated from human platelet-rich plasma. Fluorescence reflectance imaging results showed that 2PF has a 16-fold increase in fluorescence intensity compared to the autofluorescence of clots. The linear conjugate, 1PF, and free dye did not show appreciable fluorescence enhancement. 2PF fluorescence was also found 5.5-fold higher than that of the control probe, 2CF. Overall, our results suggest that 2PF binds tightly to GP-IIb/IIIa and potentially can be used for in vivo imaging of thrombosis. PMID- 15804183 TI - Collaborative research training. PMID- 15804184 TI - Characterization of rPEPT2-mediated Gly-Sar transport parameters in the rat kidney proximal tubule cell line SKPT-0193 cl.2 cultured in basic growth media. AB - The rat proximal kidney tubule cell line SKPT-0193 cl.2 (SKPT) expresses the di /tripeptide transporter PEPT2 (rPEPT2) and has been used to study PEPT2-mediated transport. Traditionally, SKPT cells have been cultured in growth media supplemented with epidermal growth factor (EGF), apotransferrin, dexamethasone, and insulin. It was recently demonstrated that omission of EGF from the culture media caused a drastic increase in the expression of rPEPT2. The hypothesis was therefore that the SKPT cell line might be able to differentiate and express rPEPT2 in the absence of the four agonists traditionally added. The aim of the study was thus to characterize Gly-Sar transport parameters in SKPT cells cultured in basic growth media (conventional media without added agonists). Morphology was studied using confocal laser scanning microscopy (CLSM) and immunohistochemistry. Monolayer integrity was evaluated using transepithelial electrical resistance (TEER) measurements and [(3)H]-mannitol permeabilities. Di /tripeptide transporter activity was studied using [(14)C]-glycylsarcosine ([(14)C]-Gly-Sar). SKPT cells grown in basic media for 4 days formed confluent monolayers with a TEER of 5.03 +/- 0.33 kOmega.cm(2) (n = 5). Apical Gly-Sar uptake peaked after 3-6 days in culture. Uptake at day 4 was 5.89 +/- 0.30 pmol.cm(-2).min(-1) (n = 3). Di-/tripeptide uptake displayed an optimum at approximately pH 6. Affinity values for cephalexin, kyotorphin, and delta aminolevulinic acid were comparable to those obtained in other PEPT2-expressing model systems. It can be concluded that SKPT cells grown in the absence of the agonists traditionally added to the culture media retain all necessary properties for PEPT2-mediated peptide uptake studies. Furthermore, the absence of the agonists might facilitate studies of hormonal regulation of PEPT2 expression and transport activity. PMID- 15804186 TI - Synthesis and antitumor activity of sulfur-containing 9-anilinoacridines. AB - A series of sulfur-containing 9-anilinoacridines related to amsacrine were synthesized and evaluated for their anticancer potential. Among the compounds, both diol-containing compounds, 2a and 3, were the most cytotoxic of the sulfide series against V-79 cells in vitro (IC(90) = 2.1 microM and 1.9 microM, respectively). Among the non-alkyl-substituted compounds (7-9), compounds with electron-donating substitution para to the sulfide (7 and 9) were more cytotoxic than the electron-withdrawing nitro-substituted compound 8. The limited SAR suggested the importance of hydroxyl functionality along with its location for the cytotoxicity in the series. A preliminary anticancer screening against P388 leukemia showed that 2a is highly active in vivo as well. Topoisomerase II inhibitory activity appeared to be involved in the cytotoxicity of compound 2a. Sulfoxide compound 2b, which is 6-7-fold less cytotoxic than its sulfide 2a, appears to be a potential bioreductive anticancer prodrug on the basis of its bioreductive metabolism findings. PMID- 15804185 TI - Functional activity of a monocarboxylate transporter, MCT1, in the human retinal pigmented epithelium cell line, ARPE-19. AB - The purpose of this study was to identify and characterize the functional activity of monocarboxylic acid transporter 1 (MCT1) on the human retinal pigmented epithelium (RPE) cell line, ARPE-19, and to evaluate whether the cell line can function as an in vitro screening tool for intravitreally administered drugs/prodrugs targeted to the MCT1 expressed in RPE. Uptake studies were carried out at 37 degrees C, for 30 s, with ARPE-19 cells. [(14)C]l-Lactic acid was selected as a substrate for this transporter. Uptake of [(14)C]L-lactic acid by ARPE-19 cells was found to exhibit saturable kinetics (K(m) = 3.1 +/- 0.6 mM and V(max) = 63.1 +/- 4.1 pmol/min/mg of protein). Monocarboxylic acids, such as benzoic acid, salicylic acid, and pyruvic acid, inhibited the uptake of [(14)C]L lactic acid whereas di- and tricarboxylic acids, such as phthalic, succinic, and citric acids, did not demonstrate any inhibitory effect. Uptake was stereospecific where D-lactic acid was less effective in inhibiting [(14)C]L lactic acid uptake than unlabeled L-lactic acid. ELISA indicated the expression of only MCT1, MCT4, and MCT8 isoforms by ARPE-19 cells. Increase in [(14)C]L lactic acid uptake was observed as the uptake medium pH was lowered from 7.4 to 5.0. Moreover, inhibition of [(14)C]L-lactic acid uptake was observed in the presence of the protonophore 2,4-dinitrophenol. Uptake was significantly decreased in the presence of sodium azide, ouabain, p-chloromercuribenzoic acid (pCMBA), N-ethylmaleamide, dithiothreitol, and p-chloromercuribenzene sulfonate (pCMBS). However, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and L thyroxine did not inhibit [(14)C]L-lactic acid. RT-PCR studies and sequence analysis of the PCR product confirmed the expression of MCT1 by ARPE-19 cells. Our results indicate that MCT1 is functionally active and is the only MCT isoform involved in the apical uptake of monocarboxylates by ARPE-19 cells. This cell line may thus be used as an effective screening tool for intravitreally administered drugs/prodrugs targeted toward MCT1 expressed on the RPE. PMID- 15804187 TI - Biological evaluation of polyester dendrimer: poly(ethylene oxide) "bow-tie" hybrids with tunable molecular weight and architecture. AB - High molecular weight (MW) polymers have shown promise in terms of improving the properties and the efficacy of low MW therapeutics. However, new systems that are highly biocompatible, are biodegradable, have well-defined MW, and have multiple functional groups for drug attachment are still needed. The biological evaluation of a library of eight polyester dendrimer-poly(ethylene oxide) (PEO) bow-tie hybrids is described here. The group of evaluated polymers was designed to include a range of MWs (from 20000 to 160000) and architectures with the number of PEO arms ranging from two to eight. In vitro experiments revealed that the polymers were nontoxic to cells and were degraded to lower MW species at pH 7.4 and pH 5.0. Biodistribution studies with (125)I-radiolabeled polymers showed that the high MW carriers (>40000) exhibited long circulation half-lives. Comparison of the renal clearances for the four-arm versus eight-arm polymers indicated that the more branched polymers were excreted more slowly into the urine, a result attributed to their decreased flexibility. Due to their essentially linear architecture that does not provide for good isolation of the iodinated phenolic moieties, the polymers with "two arms" were rapidly taken up by the liver. The biodistributions of two long-circulating high MW polymers in mice bearing subcutaneous B16F10 tumors were evaluated, and high levels of tumor accumulation were observed. These new carriers are therefore promising for applications in drug delivery and are also useful for improving our understanding of the effect of polymer architecture on pharmacokinetic properties. PMID- 15804188 TI - Molecular engineering of silk-elastinlike polymers for matrix-mediated gene delivery: biosynthesis and characterization. AB - The unique advantage of genetic engineering techniques for the design and development of polymers for controlled gene delivery lies in exquisite control over polymer structure. In this article we report the biosynthesis and characterization of a series of new silk-elastinlike protein polymers (SELPs), namely, SELP415K, with larger elastin blocks per monomer unit than SELP47K previously studied for matrix-mediated gene delivery. A new cloning strategy was used, where a block of eight elastin units (8E) was integrated into the existing DNA sequence of SELP47K monomer genes using appropriate restriction endonuclease recognition sites. Following random multimerization, multimer gene segments of desired size were selected, expressed, and purified on Ni-agarose columns. The molecular weight and sequence composition of the purified SELPs were determined by MALDI-TOF and amino acid analysis, respectively. The influence of structural changes on the rheological properties of the polymers was investigated. In addition, hydrogel disks were prepared from 47K and 415K-8mer polymer solutions, and the effects of cure time and environmental conditions on the hydrogel equilibrium swelling ratio as a function of polymer composition were studied. DNA sequencing and agarose gel electrophoresis confirmed the successful cloning of the monomer gene segment of SELP415K consisting of 312 bp. Random concatemerization of SELP415K monomer gene segments resulted in a library of SELP415K multimer sequences of 6, 8, and 10 repeats respectively, each yielding a polymer with exact molecular weight and sequence. Rheometric measurements showed that both complex shear modulus (G*) and gelation point were influenced by polymer composition. Equilibrium swelling studies on hydrogel disks prepared from 47K and 415K-8mer polymer solutions showed that changes in polymer composition resulted in different gelation patterns and increased sensitivity toward changes in temperature and ionic strength but not pH. Together these results demonstrate the potential of recombinant techniques in engineering polymers with defined structures which allows the study of the structural parameters affecting matrix mediated delivery of genes and bioactive agents. PMID- 15804189 TI - Biodistribution of boron compounds in an animal model of human undifferentiated thyroid cancer for boron neutron capture therapy. AB - Undifferentiated thyroid carcinoma (UTC) is a rapidly growing, highly invasive malignant tumor that currently lacks any effective treatment. Boron neutron capture therapy (BNCT) has been investigated recently for some types of tumors including glioblastoma multiforme and malignant melanoma. In previous studies we have shown the selective uptake of p-boronophenylalanine (BPA) by undifferentiated thyroid cancer cells in vitro and in vivo, as well as the histologic cure of 50% of the nude mice transplanted with human UTC cells when treated with BPA and an appropriate neutron beam. The present studies were performed to further optimize this treatment through the investigation of a boronated porphyrin, both alone and in combination with BPA. In vitro studies with cells in culture showed that BOPP (tetrakis-carborane carboxylate ester of 2,4-bis-(alpha,beta-dihydroxyethyl)-deutero-porphyrin IX) is localized intracellularly, with a highest concentration in the 11500g (mitochondrial enriched pellet) fraction. When BOPP was administered alone to NIH nude mice transplanted with UTC human cells, no significant tumor uptake or selectivity in our in vivo model was observed. In contrast, when BOPP was injected 5-7 days before BPA and the animals were sacrificed 60 min after administration of BPA, a significant increase in boron uptake by the tumor was found (38-45 ppm with both compounds vs 20 ppm with BPA alone). On day 5 the tissue boron selectivity ratios were tumor/blood approximately 3.8 and tumor/distal skin approximately 1.8. Other important ratios were tumor/thyroid approximately 6.6 and tumor/lung approximately 2.9. These results open the possibility of improving the efficacy of BNCT for the treatment of this so far "orphan" tumor. PMID- 15804190 TI - Amino acid ester prodrugs of the anticancer agent gemcitabine: synthesis, bioconversion, metabolic bioevasion, and hPEPT1-mediated transport. AB - Gemcitabine, a clinically effective nucleoside anticancer agent, is a polar drug with low membrane permeability and is administered intravenously. Further, extensive degradation of gemcitabine by cytidine deaminase to an inactive metabolite in the liver affects its activity adversely. Thus, strategies that provide both enhanced transport and high metabolic bioevasion would potentially lead to oral alternatives that may be clinically useful. The objective of this study was to evaluate whether amino acid ester prodrugs of gemcitabine would (a) facilitate transport across intestinal membranes or across cells that express hPEPT1 and (b) provide resistance to deamination by cytidine deaminase. 3' Monoester, 5'-monoester, and 3',5'-diester prodrugs of gemcitabine utilizing aliphatic (L-valine, D-valine, and L-isoleucine) and aromatic (L-phenylalanine and D-phenylalanine) amino acids as promoieties were synthesized and evaluated for their affinity and direct hPEPT1-mediated transport in HeLa/hPEPT1 cells. All prodrugs exhibited enhanced affinity (IC(50): 0.14-0.16 mM) for the transporter. However, only the 5'-L-valyl and 5'-L-isoleucyl monoester prodrugs exhibited (a) increased uptake (11.25- and 5.64-fold, respectively) in HeLa/hPEPT1 cells compared to HeLa cells and (b) chemical stability in buffers, that were comparable to valacyclovir, a commercially marketed oral amino acid ester prodrug. The widely disparate enzymatic bioconversion profiles of the 5'-L-valyl and 5'-L-isoleucyl prodrugs in Caco-2 cell homogenates along with their significant resistance to deamination by cytidine deaminase suggest that the disposition of gemcitabine following oral administration would be controlled by the rate of bioconversion following transport across the intestinal epithelial membrane. The combined results also suggest that it may be possible to modulate these characteristics by the choice of the amino acid promoiety. PMID- 15804191 TI - Voluntary orienting among children and adolescents with Down syndrome and MA matched typically developing children. AB - A forced-choice reaction-time (RT) task was used to examine voluntary visual orienting among children and adolescents with trisomy 21 Down syndrome and typically developing children matched at an MA of approximately 5.6 years, an age when the development of orienting abilities reaches optimal adult-like efficiency. Both groups displayed faster reaction times (RTs) when the target location was cued correctly than when cued incorrectly under both short and long SOA conditions, indicating intact orienting among children with Down syndrome. This finding is further evidence that the efficiency of many of the primary components of attention among persons with Down syndrome is consistent with their developmental level. PMID- 15804192 TI - Medical and behavioral symptoms as potential medication side effects in adults with developmental disabilities. AB - The incidence of medical and behavioral symptoms that could occur as side effects of psychotropic medication was assessed in a sample of 30 adults with developmental disabilities. Using a retrospective chart review method, we measured symptoms in six a priori classes of potential side effects over a 2-year period. The majority of side effects involved apparent effects of the psychotropic medications on behavior, mood, or sleep. Different patterns of changes in medication prescription, in relation to recorded side effects, were observed. Overall, there was a significant positive relation between the number of potential side effects recorded and the number of psychotropic medication changes made. Our findings suggest several patterns of side effects associated with psychotropic medication use. PMID- 15804193 TI - Etiological survey on intellectual disability in the northern Finland birth cohort 1986. AB - The etiology of intellectual disability was studied both in incident (n = 9,432) and prevalent (n = 9,351) populations in a one-year birth cohort born in Northern Finland in 1985-1986. Data from multiple sources were used to follow the children until the age of 11.5 years. Of the incident cases (n=119) with intellectual disabilities, 66.4% had etiologically biomedical associative factor. Paranatal factors were relatively fewer and prenatal more common compared with earlier studies. We found nearly double the prevalence of genetic factors leading to intellectual disabilities compared with a contemporary study from Norway. The differences between the populations, despite random variation, some dissimilarities between etiological categorization and diagnostic accuracy, are in most part due to true differences between the study populations and genetic pool. PMID- 15804194 TI - Use of an antecedent analysis and a force sensitive platform to compare stereotyped movements and motor tics. AB - Stereotyped movements displayed by 6 participants and tics displayed by 6 children were evaluated using an antecedent behavioral analysis and a force sensitive platform. We found that tics occurred more often in an alone condition when compared to high preference toy and play conditions, whereas stereotyped movements were more variable across conditions. Using temporal measures and spectral analysis data from the force sensitive platform, we found that stereotyped movements tended to be longer in duration and displayed more rhythmic qualities when compared to tics. Results indicate that stereotyped movements and tics differ both quantitatively and qualitatively and support the use of new technologies, such as force plate technology, to more accurately evaluate these differences. PMID- 15804195 TI - Evaluation of a decision-making curriculum designed to empower women with mental retardation to resist abuse. AB - The effectiveness of an abuse-prevention curriculum, designed to empower women with mental retardation to become effective decision-makers able to protect themselves against abuse was examined. Thirty-six women with mental retardation were randomly assigned to either an intervention or a control group. Results indicate that the performance of the women in the intervention group was superior to that of the women in the control group on measures of knowledge, decision making, and empowerment but not on a measure of stress management. The findings suggest that women with mental retardation can acquire effective decision-making strategies and apply them to situations of abuse in their lives. PMID- 15804196 TI - Evaluation of relations between specific antecedent stimuli and self-injury during functional analysis conditions. AB - We evaluated the influence of child location on the occurrence of self-injurious behavior (SIB) during functional analyses conducted in home settings. An initial functional analysis conducted with 2 children who had developmental disabilities showed undifferentiated patterns of SIB. Analyses of behavior during free play suggested that the child's location was an active variable for self-injury. Both children showed a decrease in self-injury when they were placed next to their parent versus being placed in a wheelchair or on the floor. A second functional analysis was conducted with child and parent on a couch. Results showed low percentages of self-injury during free play, but high percentages during other test conditions. Functional communication training programs for each child resulted in decreases in self-injury. PMID- 15804197 TI - Psychiatric inpatient admissions of adults with intellectual disabilities: predictive factors. AB - Information on admission to psychiatric inpatient units is lacking from the literature on contemporary services for people with intellectual disability and mental health needs. Here we report on predictors of admission for a cohort of 752 adults from this population living in community settings; 83 were admitted. We also report on two subsamples of this cohort for whom different psychometric measures concerned with psychiatric symptoms and behavior had been completed. Schizophrenia spectrum disorders and mild intellectual disabilities independently predicted admission for the total cohort. The presence of symptoms associated with psychosis and the presence of physical aggression predicted admission in the subsamples. PMID- 15804198 TI - Communication patterns of individuals with moderate or severe cognitive disabilities: interactions with unfamiliar partners. AB - Interactions between youth with cognitive disabilities and unfamiliar adult partners were observed during a "standard partner" condition to discern the effects of the use of an augmentative and alternative communication system on communication patterns. Thirteen individuals who used the System for Augmenting Language (SAL) were compared to individuals who do not speak and who do not have SAL experience and individuals who speak. In general, the SAL users fell in the middle of the range, communicating better than the symbol-naive nonspeakers, yet not quite as well as the natural speakers with cognitive disabilities. These findings highlight the distinct contributions symbol experience makes to communication interactions with unfamiliar partners. PMID- 15804200 TI - Challenges facing employers in the treatment of depression. AB - OBJECTIVE: To review the challenges of treating depression in the workplace in the environment of increased health care costs. SUMMARY: The cost of health care is on the rise, and employers and employees are at odds over who will share the burden of these costs. The focus of cost containment has been to minimize drug cost; however, employers need to be aware of indirect costs of medical illness. Due to its prevalence and rate of undertreatment, depression is one of the main disease states that employers should target in their disease management efforts. CONCLUSION: In order to treat depression appropriately, we must consider the social stigma, treatment barriers, and health care structure that exist to treat depression. Additional outcomes data is needed to demonstrate to employers, the largest purchasers of health care, the benefit of managing depression in the workplace. PMID- 15804201 TI - Improving depression treatment by integrated care. AB - OBJECTIVE: To identify the clinical and economic implications of depression in the workplace and review how integrated care models can improve overall patient outcomes. SUMMARY: Depression is a significant financial burden to the employer due to lost days of work and decreased productivity. Employers are demanding return on the investment for their increasing health care expenditures. The cost of depression to employers may be contained by delivering care using integrated models that leverage primary care provider treatment with care management and mental health consultation. CONCLUSION: There is a need to reduce silos in the organization and financing of mental health care to prevent cost shifting that provides no benefit to patients, payers or providers. Poor mental health care will likely lead to a rise in absenteeism and presenteeism. PMID- 15804202 TI - NCQA behavioral health measurement efforts. AB - OBJECTIVE: To review the role of the National Committee for Quality Assurance (NCQA) in ensuring the quality of care in the managed care setting and identify novel strategies to improve performance rates for Health Plan Employer Data and Information Set (HEDIS) measures, particularly in the area of depression. SUMMARY: NCQA, by regulating HEDIS measures, sets the standards by which managed care organizations evaluate their performance in providing care for their enrollees. The medication management measure for depression evaluates practitioner contacts and acute and continuation phase treatments for persons treated with an antidepressant. Despite increased detection and management of patients with depression, there is still room for improvement in HEDIS performance rates for this chronic disease. CONCLUSION: NCQA hopes to improve collaboration among managed care organizations and managed behavioral health organizations In addition, NCQA regularly reevaluates the HEDIS measures using input from panels of experts. Incentive p r o grams for providers who deliver quality care may also help to improve HEDIS performance rates for depression. Research is under way to evaluate the feasibility and re t u rn on investment for pay-for-performance programs in depression. PMID- 15804203 TI - Promoting health and productivity for depressed patients in the workplace. AB - OBJECTIVE: To discuss the impact of major disease states, including depression, in the loss of productivity in the workplace and how integration of health care can decrease cost to employers. SUMMARY: The majority of costs associated with depressive illness can be traced to lost productivity, and the employer, therefore, bears most of the economic burden. Efforts to improve employee health and productivity have been hampered by the compartmentalization of medical costs, pharmacy costs, behavioral health costs, and productivity measures. This situation can be rectified by "busting" these silos and promoting a reintegration of prospective costs and parties. Health risk assessments enable employers to identify illnesses that are suitable targets for integrated health and productivity management programs. In the case of depression, employers can act proactively to identify employees at risk, working to minimize risk factors such as stress before these individuals become heavy utilizers of company resources. For employees who are currently depressed, recent research evidence has demonstrated that pharmacotherapy can have a dramatic and positive effect on lost productivity, absenteeism, and presenteeism. The selection of antidepressants and subsequent follow-up must be improved, however, if the benefits of pharmacotherapy are to be optimized. CONCLUSION: Understanding the linkage of disease management and productivity in the workplace can result in dramatic decreases in absenteeism and presenteeism and increased cost savings to the employers. PMID- 15804204 TI - Depression management in the workplace: a case study. AB - OBJECTIVE: To review a case of a depression management program at PPG Industries with the potential to improve outcomes on functioning and productivity in the workplace. SUMMARY: The need for major improvements in depression management has been well established. While most in the corporate world are aware of these deficiencies, the perception is that this fragmented system is difficult to change. PPG Industries, a medium-sized, Fortune 500 company manufacturing paints, stains, and sealants, has launched a successful and comprehensive program in the workplace that has improved outcomes for depressed employees. The PPG approach is practical, addressing each step in the process one at a time. This process began by establishing a close working relationship between all entities responsible for employee health, many of which are currently carved out. This enabled the company to have a comprehensive view of their depressed population and to examine their functional outcomes as well. In an effort to help identify depressed employees, the company educated their physicians and care managers and launched a confidential Web site to reduce mental health stigma and aid in employee screening and education. A multidisciplinary team was then assembled to facilitate the treatment of these patients. All of these steps were measurable, and the preliminary results of this process are summarized. CONCLUSION: A disease management program that incorporates a multifaceted, collaborative approach to treat depression is feasible and may improve care and decrease cost to employers. PMID- 15804205 TI - Effect of prescriber education on the use of medications contraindicated in older adults in a managed medicare population. AB - OBJECTIVE: To measure the effect of pharmacy intervention programs on the percentage of older adult members filling prescriptions for one or more contraindicated medications in a managed Medicare population. METHODS: Prescribers of potentially inappropriate (i.e., contraindicated) medications in a managed Medicare + Choice (now known as Medicare Advantage) health maintenance organization (HMO) were identified from drug claims each calendar quarter for letter and telephone contact. The initial study population was composed of members in the Medicare + Choice HMO who were aged 65 years or older and who utilized their managed care pharmacy benefit during the 4-year measurement period from 1999 fourth quarter (Q4) through 2003 Q4. The study population was expanded in 2003 Q1 to include all patients aged 65 years or older, including commercial HMO members and Medicare + Choice members. The prescribers of target (contraindicated) drugs were contacted by a letter that described the program and included a prescriber-specific list of patients who had received a potentially inappropriate drug. Each prescriber was requested to review the report and change the contraindicated drug to a more appropriate agent, lower the medication dose, or carefully monitor those patients who could not discontinue the drug. A clinical pharmacist contacted high-volume prescribers of target drugs (4 or more patients in a given quarter) by telephone to discuss the identified patients. RESULTS: The incidence of inappropriate use of the target medications in older adults declined by 19%, from an absolute 5.3% of prescriptions in the base period (1999 Q4) to 4.3% at the end of the first 2-year period, in 2001 Q4. A change in the target drugs in 2002 Q1 from a list based upon the Beers' criteria to a list based in part on the Zhan et al. criteria was associated with a 45% reduction in the percentage (2.4%) of older adult members who received target drugs; this proportion remained essentially unchanged at 2.2% through 2003 Q4, the end of the second half of the 4-year intervention period. CONCLUSIONS: A clinical intervention program targeting prescribers of drugs judged to be contraindicated in older adults was associated with a decline in the percentage of HMO members receiving one or more target contraindicated drugs. After a 4-year intervention period, 2.2% of older adult HMO members continued to receive one or more target contraindicated drugs. PMID- 15804206 TI - Comparison of mental health resources used by patients with bipolar disorder treated with risperidone, olanzapine, or quetiapine. AB - OBJECTIVE: The atypical antipsychotics, risperidone, olanzapine, and quetiapine, have been approved by the U.S. Food and Drug Administration for treatment of mania associated with bipolar disorder. Information on the relative mental health resource use of these therapies is helpful to pharmacy managers since differences in efficacy and safety may translate into differences in mental health care utilization. We compared charges for other mental health services associated with risperidone, olanzapine, and quetiapine treatment of patients with bipolar disorder to assess whether there were significant differences between these therapies. A secondary analysis involved dose-equivalent adjustment of the average allowed charge of the 3 atypical antipsychotics. METHODS: This was a retrospective study based on administrative data for 46 U.S. commercial health plans represented in a commercial database covering the period January 1998 through April 2002. The 6,625 patients included in the study had at least 2 contiguous pharmacy claims for a study antipsychotic, had received no other antipsychotics concurrently, and had not switched from an alternative antipsychotic in the preceding 90 days. Provider-submitted (billed) charges were selected in preference to paid amounts as being more accurate indicators of relative differences in the use of mental health resources. Mental health care charges were measured per patient per month (PPPM) and included charges for the study antipsychotics and charges for the other mental health care services (inpatient, physician and other ambulatory, and other psychotropic medications). Differences in other mental health care charges PPPM among the 3 therapies were assessed with multivariate regression, adjusting for differing patient characteristics. Differences in antipsychotic drug charges PPPM were assessed after adjustment to reflect an equivalent average daily dose. RESULTS: Regression estimates adjusted for patient differences did not show statistically significant differences in other mental health care charges PPPM among the 3 antipsychotic drug therapies. Other mental health charges associated with quetiapine were estimated to be 14 US dollars, or 3% lower than those associated with risperidone, but this difference was not statistically significant (P = 0.069). The PPPM charges for quetiapine versus olanzapine and olanzapine versus risperidone were also not different (P = 0.231 and P = 0.39, respectively). After adjusting for differences in average daily dose, risperidone and quetiapine had antipsychotic drug charges that were 84 US dollars and 76 US dollars PPPM lower than those of olanzapine (P < 0.01); the difference between the adjusted drug charges PPPM for risperidone and quetiapine was not significant. CONCLUSION: Total charges for mental health services other than the study drug were not different for risperidone, olanzapine, and quetiapine in patients treated for bipolar disorder. However, based on prescription charges, olanzapine appears to be considerably more costly at an equivalent daily dose than either risperidone or quetiapine. PMID- 15804207 TI - Drug persistency patterns for patients treated with rivastigmine or donepezil in usual care settings. AB - OBJECTIVE: To compare levels of persistency with 2 cholinesterase (ChE) inhibitors--rivastigmine and donepezil--for the treatment of Alzheimer's disease (AD) through the use of administrative claims data. METHODS: This retrospective cohort study identified treatment-naive, community-based AD patients having an initial prescription (index event) for rivastigmine or donepezil between June and December 2000, in the United States, from pharmacy claims in a proprietary administrative claims database. Patients were excluded if they received either drug during the 180 days prior to their index prescription or if they did not have continuous plan enrollment during this period and for at least 90 days following the index date. The probability of treatment discontinuation within the first 60 days of treatment was estimated. Time to treatment discontinuation was analyzed for the cohort of patients that remained on therapy > or =60 days as well as for subgroups of the cohort reaching either approved or maximum recommended doses of donepezil or rivastigmine. Treatment discontinuation was defined as either a stop of therapy (no prescription refill within 60 days of estimated completion of prior prescription) or a switch to an alternative AD drug. Kaplan-Meier survival and proportional hazard model analyses were performed. Proportion of days covered (PDC) by an AD therapy was also evaluated in each quarter during the first year of follow-up. RESULTS: Of the newly treated AD study population, 30.4% (171/563) of rivastigmine patients and 31.2% (583/1,871) of donepezil patients discontinued treatment within 60 days of starting therapy (P = 0.72). For the cohort of patients that remained on therapy > or =60 days, the mean time to treatment discontinuation was 331 days (95% confidence interval [CI], 307-355) for rivastigmine patients (n = 392) versus 337 days (95% CI, 322-352) for donepezil patients (n = 1288). The proportion of patients with a PDC > or =80% after 12 months of follow-up was 23% for the donepezil group and 19% for the rivastigmine group (P = 0.34). For the cohort subgroup that reached an approved dose, the mean time to treatment discontinuation was 346 days (95% CI, 318-374) for rivastigmine patients (n = 282) versus 338 days (95% CI, 323-353) for donepezil patients (n = 1,283). For the cohort subgroup that reached the maximum recommended dose, the mean time to treatment discontinuation was 396 days (95% CI, 343-449) for rivastigmine patients (n = 61) versus 364 days (95% CI, 344-384) for donepezil patients (n = 712). CONCLUSION: Newly treated AD patients in a usual care setting who initiate therapy with either rivastigmine or donepezil have similar levels of persistency with treatment. PMID- 15804208 TI - Regression methods in the empiric analysis of health care data. AB - OBJECTIVE: The aim of this paper is to provide health care decision makers with a conceptual foundation for regression analysis by describing the principles of correlation, regression, and residual assessment. SUMMARY: Researchers are often faced with the need to describe quantitatively the relationships between outcomes and predictors , with the objective of explaining trends, testing hypotheses, or developing models for forecasting. Regression models are able to incorporate complex mathematical functions and operands (the variables that are manipulated) to best describe the associations between sets of variables. Unlike many other statistical techniques, regression allows for the inclusion of variables that may control for confounding phenomena or risk factors. For robust analyses to be conducted, however, the assumptions of regression must be understood and researchers must be aware of diagnostic tests and the appropriate procedures that may be used to correct for violations in model assumptions. CONCLUSION: Despite the complexities and intricacies that can exist in regression , this statistical technique may be applied to a wide range of studies in managed care settings. Given the increased availability of data in administrative databases, the application of these procedures to pharmacoeconomics and outcomes assessments may result in more varied and useful scientific investigations and provide a more solid foundation for health care decision making. PMID- 15804209 TI - Development of a complementary and alternative medicine (CAM) pharmacy and therapeutics (P&T) subcommittee and CAM guide for providers. AB - OBJECTIVE: The objective was 2-fold: (1) to evaluate the feasibility and value of developing a Pharmacy and Therapeutics (P&T) subcommittee aimed at scientifically evaluating complementary and alternative medicine (CAM) products for an integrated managed care organization (IMCO) and (2) to assess provider acceptance and usefulness of a CAM guide. METHODS: Three factors drove the decision to form a CAM P&T subcommittee to evaluate current commonly used CAM products: (1) physicians, pharmacists, and dieticians expressed a desire for an easy-to-use, scientifically based mechanism for evaluating the ever-increasing number of CAM products; (2) Intermountain Health Care Health Plans (Health Plans), the insurance division of this IMCO, offers access to certain CAM products to its members at a discounted price in an effort to remain competitive with other IMCOs; and (3) this IMCO owns and operates more than a dozen community pharmacies that sell CAM products. Some IMCO clinicians believed an efficacy and safety review of the products offered through the organization was warranted. Subcommittee members included clinical pharmacists (IMCO and university), pharmacy directors, a community pharmacist, practicing physicians (from the drug P&T committee), a medical director, dieticians and nutritionists, and a representative from the Health Plans sales department. The primary outcome was the development of a CAM guide listing recommendations for use of CAM products. Outcome measures included survey results (survey sent with guide to physicians and (pharmacists) regarding acceptance and usefulness of the guide. RESULTS: The CAM P&T subcommittee met monthly to evaluate current commonly used CAM products. A CAM guide was developed in paperback and electronic versions. The electronic version was downloadable to handheld devices. Thousands of CAM guides were disseminated to IMCO-employed physicians, network pharmacies, dieticians, and nutritionists affiliated with this managed care organization. A survey that accompanied distribution of the first CAM guide in 2003 showed that 89% of physicians and pharmacists felt that the guide would be somewhat or very helpful as a counseling aide; the remainder was unsure. A second CAM guide was disseminated one year later, in 2004. The accompanying survey showed that 78% of physicians and 97% of pharmacists felt that the guide would be somewhat or very helpful as a counseling aide; 7% of physicians and 3% of pharmacists felt the guide would be unhelpful. CONCLUSION: A CAM guide developed through the work of a subcommittee of the P&T committee of this IMCO appears to be widely accepted by pharmacists and physicians. A CAM guide should be easy to use and available online with the ability to download to a handheld device. PMID- 15804210 TI - Evidence-based medicine: which drugs are truly contraindicated for use in older adults? PMID- 15804211 TI - Does persistence with drugs for Alzheimer's disease matter? PMID- 15804212 TI - The fourth EC report on laboratory animal use--is it sufficiently revealing? PMID- 15804213 TI - Lack of predictivity of the rat lethality (LD50) test for ecological and human health effects. AB - The relationship between acute toxicity in rats (LD50 values) and indicators of potential health hazards in humans was investigated, based on a chemical population-based paradigm (i.e. the "chemical diversity approach"). These structure-activity relationship-based analyses indicate that high toxicity in rats (i.e. a low LD50 value) is not a good predictor of health effects in humans. In fact, it was found that high acute toxicity to minnows, as well as toxicity to cultured cells, showed significantly greater associations with the potential for health effects than rat LD50 values. PMID- 15804214 TI - Ecotoxicological evaluation of municipal sludge. AB - Municipal wastes originating from urban and industrial areas have become a major source of soil, ground and surface water pollution. These undesirable agents in our environment significantly interact with our flora and fauna. The aim of this study was to test samples of municipal sludge (MS) for their ecotoxicological potential by using sensitive bioassays involving a plant, Vicia faba, and the earthworm, Eisenia foetida. A 10% leachate of MS was prepared for the experiments, and V. faba seedlings were exposed to three leachate concentrations (2.5%, 5% and 10%) for 5 days. The findings revealed chromosome aberrations during the metaphase as well as the anaphase of cell division, and inhibition of the mitotic index, which reflects that MS originating from domestic and other human activities may be genotoxic to the living organisms of the ecosystem. Abnormalities in chlorophyll content, plant growth, root length, shoot length and root/shoot length ratio in V. faba clearly indicated the toxicity of the sludge. Behavioural and reproduction studies with E. foetida also provided evidence for the toxic nature of the MS. PMID- 15804215 TI - Human research tissue banks: the ATRA Project for establishing a human research tissue bank in switzerland. AB - A large number of experiments in biomedical research are carried out on tissues, but, even though the results should be applicable to humans, these tissues are mainly of animal origin. The difficulty encountered in obtaining human organs and tissues is an acknowledged problem: not enough human tissues are available to meet research needs. We are introducing the ATRA Project, with the purpose of supporting progress in biomedical research in Switzerland through the establishment of one or more human tissue banks, which will be able to find, treat, preserve and supply human material. Where similar projects have already been launched, concerns have been expressed that donation for research purposes might compete with donation for transplantation, but most organs and tissues are in any case non-transplantable. Surplus surgical tissue is considered "sanitary waste", and must be treated according to specific regulations for collection, packaging, transport, treatment and disposal. A human tissue bank would not only abate the costs of treating sanitary waste, but would actually turn what is now considered waste into a resource which could be used to save human and animal lives. PMID- 15804218 TI - Who will do general surgery? AB - Advantages to patients of a single anaesthetic for more than one operation are obvious; attracting generalist surgeons, training them and ensuring they have adequate credentials remain hurdles. PMID- 15804219 TI - Child pedestrian safety: the role of behavioural science. AB - Environmental strategies must be complemented by behavioural approaches to help children learn to use roads safely. PMID- 15804220 TI - Riluzole: a glimmer of hope in the treatment of motor neurone disease. AB - Early experience confirms that riluzole improves survival and is well tolerated. PMID- 15804221 TI - Access to medicines and high-quality therapeutics: global responsibilities for clinical pharmacology. AB - A major theme of the 2004 World Congress of Clinical Pharmacology and Therapeutics was worldwide equity of access to medicines. PMID- 15804222 TI - Variations in indicated care of patients with acute coronary syndromes in Queensland hospitals. AB - OBJECTIVE: To identify variation in the rates of use of key evidence-based therapies and in clinical outcomes among patients hospitalised with acute coronary syndromes (ACS). DESIGN: Retrospective analysis of data on care processes and clinical outcomes of representative patient samples recorded by the Queensland Health Cardiac Collaborative registry. SETTING: 18 public hospitals (3 tertiary, 15 non-tertiary) in Queensland, August 2001 to December 2003. STUDY POPULATION: 2156 patients who died or were discharged after troponin-positive ACS. MAIN OUTCOME MEASURES: Comparison of proportions of highly eligible patients receiving indicated care and in-hospital mortality between subgroups categorised by age, sex, comorbidities (diabetes, renal failure, chronic obstructive pulmonary disease and mental disorder), type of admitting hospital (tertiary or non-tertiary), and cardiologist involvement (transfer or non-transfer to cardiology unit). RESULTS: Patients aged > or = 65 years were less likely than younger patients to receive heparin (79% v 87%), beta-blockers (79% v 87%), lipid lowering agents (78% v 87%), coronary angiography (51% v 66%), and referral to cardiac rehabilitation (17% v 33%). Patients with diabetes were less likely than others to receive coronary angiography (50% v 63%), while those with moderate to severe renal failure were less likely to receive thrombolysis (52% v 84%), heparin (71% v 83%), beta-blockers (69% v 84%), lipid-lowering agents (61% v 84%), in-hospital cardiac counselling (46% v 64%) and referral to cardiac rehabilitation (9% v 25%). Patients admitted to tertiary hospitals were more likely than those admitted to non-tertiary hospitals to receive coronary angiography (85% v 55%) and referral to cardiac rehabilitation (36% v 21%). Risk adjusted mortality was highest in patients with moderate to severe renal failure (15% v 3%) and older patients (6% v 2%). CONCLUSIONS: Variations exist in the provision of indicated care to patients with ACS according to age, diabetic status, renal function and type of admitting hospital. Excess mortality in elderly patients and in those with advanced renal disease may be partially attributable to failure to use key therapies. PMID- 15804223 TI - Hypertensive disorders in pregnancy: a population-based study. AB - OBJECTIVES: To determine population-based rates and outcomes of hypertensive disorders in pregnancy. DESIGN: Cross-sectional study using linked population databases. SETTING AND PARTICIPANTS: All women, and their babies, discharged from hospital following birth in New South Wales, between 1 January 2000 and 31 December 2002. MAIN OUTCOME MEASURES: Rates of hypertensive disorders in pregnancy, maternal and infant morbidity and mortality, and level of hospital care for the birth admission. RESULTS: 250 173 women and their 255 931 infants were included in the study. Overall, 24 517 women (9.8%) had a hypertensive disorder in pregnancy, including 1411 (0.6%) with chronic hypertension, 10 379 (4.2%) with pre-eclampsia, 731 (0.3%) with chronic hypertension with superimposed pre-eclampsia, and 10 864 (4.3%) with gestational hypertension. Women with, and infants exposed to, hypertension were more likely to suffer death or major morbidity than those without hypertension. Infants of mothers with hypertension were more likely to be to born preterm and small for gestational age. Just over half the women with major morbidity or mortality delivered in hospitals with a high level of medical care. In contrast, most infants with major morbidity or mortality were delivered in hospitals with neonatal intensive care units. CONCLUSIONS: Hypertension is a common complication of pregnancy, and adverse outcomes are increased among hypertensive women and their babies. Clinicians appear to be better at identifying and seeking an appropriate level of care for pregnancies where the infant is at risk of a poor outcome than when the mother is at risk. More specific antenatal indicators of poor maternal outcome would help guide the referral of hypertensive women to higher levels of care. PMID- 15804224 TI - Multispecialty surgical conditions in general practice. AB - OBJECTIVES: To report the incidence of multispecialty surgical conditions in patients presenting to a procedural general practice. DESIGN AND SETTING: A more than 18-year survey (1 August 1983 - 31 January 2002) of the surgical records of a general practitioner-surgeon in an urban general practice. PARTICIPANTS: 211 patients each with multiple, elective, surgical problems (mostly non-major) treated at one operation. RESULTS: The 211 patients represented 9.03% of the practitioner's elective, non-referred, general practice surgical workload. Two separate procedures were performed at one surgical episode for 155 patients (73.5%), three separate procedures for 53 patients (25.1%), and four separate procedures for three patients (1.4%). Having all surgical conditions treated in a single episode resulted in considerable savings in time, convenience and expense for both the patient and the health care system. CONCLUSION: There appears to be a place, at least in our major cities, for an appropriately trained and recognised general surgeon, to service patients with more than one minor condition requiring surgery. PMID- 15804225 TI - "Operation South East Asia Tsunami Assist": an Australian team in the Maldives. PMID- 15804226 TI - Difficulties in provision of bariatric surgical services to the morbidly obese. AB - Morbid obesity (defined as having a body mass index [BMI] > 40 kg/m(2), or BMI > 35 kg/m(2) with obesity-related comorbidities) is a medical disorder associated with increased morbidity and mortality. Management guidelines published by the National Health and Medical Research Council and by similar US and UK bodies have recommended surgery as the most effective treatment available for selected patients with morbid obesity. A recent meta-analysis of obesity surgery has documented its safety and effectiveness in resolving some of the major medical comorbidities that occur in obese patients. To date, no intervention other than surgery has proven either effective or cost-effective in treating severe obesity and its associated medical conditions. Targeting patients with metabolic complications of obesity (eg, type 2 diabetes) could lead to substantial cost savings for the public health system. Currently, Medicare pays for privately insured patients to undergo obesity surgery, while uninsured patients are denied access to surgery in public hospitals. This raises significant equity issues that should be addressed. PMID- 15804227 TI - Lymphomatous infiltration of the peripheral nervous system in enteropathy associated T-cell lymphoma. PMID- 15804228 TI - Life-threatening milk-alkali syndrome resulting from antacid ingestion during pregnancy. PMID- 15804229 TI - 7. Language disorders and autism. AB - Early diagnosis of language disorders and autism is important, and early intervention for autism and some language disorders makes a difference. Developmental surveillance of children to detect these disorders should be a routine part of medical practice. The persistence and pervasiveness of communication and socialising deficits differentiate children with autism from those with specific developmental language disorders. Hearing and vision assessment is essential in any communication disorder. Interventions, targeted to identified areas of need, should encompass communication enhancement, behavioural therapy, educational modification, parent education and family support. Pharmacological interventions have an important but discrete role in autism, but there are no magic bullets. It is important to remember that the normal childhood illnesses occur in children with developmental disorders. Parents should be directed to reliable websites on the Internet, and given information and books to read as well as phone numbers of relevant services (eg, autism associations). There is a need for increased government financial support for early intervention programs. PMID- 15804230 TI - Advances in childhood leukaemia. PMID- 15804231 TI - Tsunami lung: a necrotising pneumonia in survivors of the Asian tsunami. PMID- 15804232 TI - Warfarin reversal: consensus guidelines, on behalf of the Australasian Society of Thrombosis and Haemostasis. PMID- 15804233 TI - X-ray machine assaults anaesthetist. PMID- 15804234 TI - Transdermal delivery of interferon-gamma (IFN-gamma) mediated by penetratin, a cell-permeable peptide. AB - IFN-gamma (interferon-gamma) has several applications in the treatment of IFN gamma-related skin disorders. While systemic delivery - the major route used to administer IFN-gamma - results in significant side effects and toxicity, including fever, fatigue, nausea, vomiting and neurotoxicity, transdermal delivery has a very low transduction efficiency. In order to improve the efficiency of transdermal IFN-gamma delivery, we introduced a Pen (penetratin) peptide, a 16-amino-acid-long polypeptide corresponding to the third helix of the DNA-binding domain (homoeodomain) of Antennapedia (a Drosophila transcription factor). The human IFN-gamma gene was then fused with a gene fragment that encodes the Pen of Antennapedia in a bacterial expression vector, producing a genetic in-frame Pen-IFN-gamma. The expressed and purified Pen-IFN-gamma was then found to have a much more efficient transduction profile than native IFN-gamma. In addition, compared with native IFN-gamma, Pen-IFN-gamma exhibited similar activities when added exogenously to a culture medium: (i) induction of IRF-1 gene expression, and (ii) NF-kappaB (nuclear factor kappaB) luciferase reporter activation. These results indicate that the transdermal delivery system using Pen may be an excellent way to replenish IFN-gamma in the various disorders related to this cytokine. PMID- 15804235 TI - Enzymatic properties of native and deglycosylated hybrid aspen (Populus tremulaxtremuloides) xyloglucan endotransglycosylase 16A expressed in Pichia pastoris. AB - The cDNA encoding a xyloglucan endotransglycosylase, PttXET16A, from hybrid aspen (Populus tremulaxtremuloides) has been isolated from an expressed sequence tag library and expressed in the methylotrophic yeast Pichia pastoris. Sequence analysis indicated a high degree of similarity with other proteins in the XTH (xyloglucan transglycosylase/hydrolase) gene subfamily of GH16 (glycoside hydrolase family 16). In addition to the conserved GH16 catalytic sequence motif, PttXET16A contains a conserved N-glycosylation site situated proximal to the predicted catalytic residues. MS analysis indicated that the recombinant PttXET16A expressed in P. pastoris is heterogeneous due to the presence of variable N-glycosylation and incomplete cleavage of the alpha-factor secretion signal peptide. Removal of the N-glycan by endoglycosidase H treatment did not influence the catalytic activity significantly. Similarly, site-directed mutagenesis of Asn93 to serine to remove the N-glycosylation site resulted in an enzyme which was comparable with the wild-type enzyme in specific activity and thermal stability but had clearly reduced solubility. Hydrolytic activity was detected neither in wild-type PttXET16A before or after enzymatic deglycosylation nor in PttXET16A N93S (Asn93-->Ser) mutant. PMID- 15804236 TI - Characterization of mouse amino acid transporter B0AT1 (slc6a19). AB - The mechanism of the mouse (m)B0AT1 (slc6a19) transporter was studied in detail using two electrode voltage-clamp techniques and tracer studies in the Xenopus oocyte expression system. All neutral amino acids induced inward currents at physiological potentials, but large neutral non-aromatic amino acids were the preferred substrates of mB0AT1. Substrates were transported with K0.5 values ranging from approx. 1 mM to approx. 10 mM. The transporter mediates Na+-amino acid co-transport with a stoichiometry of 1:1. No other ions were involved in the transport mechanism. An increase in the extracellular Na+ concentration reduced the K0.5 for leucine, and vice versa. Moreover, the K0.5 values and Vmax values of both substrates varied with the membrane potential. As a result, K0.5 and Vmax values are a complex function of the concentration of substrate and co-substrate and the membrane potential. A model is presented assuming random binding order and a positive charge associated with the ternary [Na+-substrate-transporter] complex, which is consistent with the experimental data. PMID- 15804237 TI - Highly conserved proximal promoter element harbouring paired Sox9-binding sites contributes to the tissue- and developmental stage-specific activity of the matrilin-1 gene. AB - The matrilin-1 gene has the unique feature that it is expressed in chondrocytes in a developmental stage-specific manner. Previously, we found that the chicken matrilin-1 long promoter with or without the intronic enhancer and the short promoter with the intronic enhancer restricted the transgene expression to the columnar proliferative chondroblasts and prehypertrophic chondrocytes of growth plate cartilage in transgenic mice. To study whether the short promoter shared by these transgenes harbours cartilage-specific control elements, we generated transgenic mice expressing the LacZ reporter gene under the control of the matrilin-1 promoter between -338 and +67. Histological analysis of the founder embryos demonstrated relatively weak transgene activity in the developing chondrocranium, axial and appendicular skeleton with highest level of expression in the columnar proliferating chondroblasts and prehypertrophic chondrocytes. Computer analysis of the matrilin-1 genes of amniotes revealed a highly conserved Pe1 (proximal promoter element 1) and two less-conserved sequence blocks in the distal promoter region. The inverted Sox motifs of the Pe1 element interacted with chondrogenic transcription factors Sox9, L-Sox5 and Sox6 in vitro and another factor bound to the spacer region. Point mutations in the Sox motifs or in the spacer region interfered with or altered the formation of nucleoprotein complexes in vitro and significantly decreased the reporter gene activity in transient expression assays in chondrocytes. In vivo occupancy of the Sox motifs in genomic footprinting in the expressing cell type, but not in fibroblasts, also supported the involvement of Pe1 in the tissue-specific regulation of the gene. Our results indicate that interaction of Pe1 with distal DNA elements is required for the high level, cartilage- and developmental stage-specific transgene expression. PMID- 15804238 TI - Topology of transmembrane segments 1-4 in the human chloride/bicarbonate anion exchanger 1 (AE1) by scanning N-glycosylation mutagenesis. AB - Human AE1 (anion exchanger 1), or Band 3, is an abundant membrane glycoprotein found in the plasma membrane of erythrocytes. The physiological role of the protein is to carry out chloride/bicarbonate exchange across the plasma membrane, a process that increases the carbon-dioxide-carrying capacity of blood. To study the topology of TMs (transmembrane segments) 1-4, a series of scanning N glycosylation mutants were created spanning the region from EC (extracellular loop) 1 to EC2 in full-length AE1. These constructs were expressed in HEK-293 (human embryonic kidney) cells, and their N-glycosylation efficiencies were determined. Unexpectedly, positions within putative TMs 2 and 3 could be efficiently glycosylated. In contrast, the same positions were very poorly glycosylated when present in mutant AE1 with the SAO (Southeast Asian ovalocytosis) deletion (DeltaA400-A408) in TM1. These results suggest that the TM2-3 region of AE1 may become transiently exposed to the endoplasmic reticulum lumen during biosynthesis, and that there is a competition between proper folding of the region into the membrane and N-glycosylation at introduced sites. The SAO deletion disrupts the proper integration of TMs 1-2, probably leaving the region exposed to the cytosol. As a result, engineered N-glycosylation acceptor sites in TM2-3 could not be utilized by the oligosaccharyltransferase in this mutant form of AE1. The properties of TM2-3 suggest that these segments form a re-entrant loop in human AE1. PMID- 15804239 TI - Brain MRI in late-onset multiple sclerosis. AB - Multiple sclerosis (MS) with clinical onset after 50 years of age is unusual (between 1 and 6%) and is frequently misdiagnosed. Furthermore, brain magnetic resonance imaging (MRI) abnormalities are frequently observed in subjects over 50 years of age. The aim of this study was to describe brain MRI in late-onset MS to evaluate the sensitivity and specificity of radiological MS criteria in patients aged over 50 years. We evaluated the brain MRI of 20 patients with onset of MS after 50 years of age. We compared these MRI with 26 controls matched for age, sex and vascular risk factors. MRI were blindly analysed by two neuroradiologists according to Paty et al.'s [Neurology38 (1988) 180] criteria, Fazekas et al.'s [Neurology38 (1988) 1822] criteria and Barkhof et al.'s [Brain120 (1997) 2059] criteria. The mean age at MRI scanning was 58 years. Sensitivity was 90% for Paty et al.'s criteria, 80% for Fazekas et al.'s criteria and 85% for Barkhof et al.'s criteria. Specificity was 54% for Paty et al.'s criteria, 69% for Fazekas et al.'s criteria and 65% for Barkhof et al.'s criteria. Barkhof et al.'s criteria are less specific in older patients than in young patients. We suggest that spinal cord MRI and cerebrospinal fluid analysis should be systematically performed in suspected late-onset MS in order to increase the specificity of the diagnosis. PMID- 15804240 TI - The epidemiology of epilepsy in Europe - a systematic review. AB - Population-based epidemiological studies on epilepsy are available mainly from the UK and the Nordic, Baltic and western Mediterranean countries. No studies were identified from large areas of Europe, especially from the former eastern Europe (except the Baltic countries) and the eastern Mediterranean countries. Based on the prevalence of epilepsy in different studies and accounting for incomplete case identification the estimated number of children and adolescents in Europe with active epilepsy is 0.9 million (prevalence 4.5-5.0 per 1000), 1.9 million in ages 20-64 years (prevalence six per 1000) and 0.6 million in ages 65 years and older (prevalence seven per 1000). Approximately 20-30% of the epilepsy population have more than one seizure per month. Based on the age-specific incidence rates in European studies, the estimated number of new cases per year amongst European children and adolescents is 130,000 (incidence rate 70 per 100,000), 96,000 in adults 20-64 years (incidence rate 30 per 100,000) and 85,000 in the elderly 65 years and older (incidence 100 per 100,000). The proportion of both new and established cases with epilepsy in the young, adults and elderly in individual countries may differ substantially from total European distribution because of differences in age structure. PMID- 15804241 TI - Cost-effectiveness of 18F-fluorodeoxyglucose positron emission tomography in the assessment of early dementia from a Belgian and European perspective. AB - (18)F-fluoro-deoxyglucose positron emission tomography (FDG PET) can aid to predict AD in an early stage. The aim of this study was to estimate the economic effects of incorporating FDG PET in the diagnostic work-up of AD in a Belgian and European setting. A decision tree analysis was followed comparing a conventional algorithm using diagnostic clinical criteria and one that also incorporates PET. Major outcome terms were overall cost per patient in either strategy; diagnostic accuracy and cost per accurate diagnosis. A sensitivity analysis was performed for four critical variables: cost of PET, sensitivity and specificity of PET and delay in cognitive decline because of appropriate medication. Cost-savings per accurate diagnosis ranged from 623-6110 Euro in favour of the proposed algorithm with PET. For the same cost, more accurate diagnoses were made, resulting in benefit for patients and society. The positive results were maintained over a wide range of values for the critical variables and were expandable to other European countries with a similar health system. Therefore, incorporation of FDG PET into the clinical diagnostic work up of patients with early symptoms of cognitive decline can be advocated. PMID- 15804242 TI - Care needs and economic consequences after acute ischemic stroke: the Erlangen Stroke Project. AB - The objective was to determine the functional outcome, location of care and economic consequences in the first 3 months after ischemic stroke. As part of the Erlangen Stroke Project, (ESPro) information was collected on patients suffering a first-ever ischemic stroke. Three months after the stroke, location of care, dependence on caregivers and function based on Barthel Index: poor (0-55), moderate (60-90) or good function (95-100) were recorded. Data about health services used were combined with cost estimates for Germany (2000 Euros, undiscounted). Of 491 patients hospitalized, 383 were alive 3 months afterwards, 79% residing in the community. The majority of patients with poor function (60%) were still in institutional care. Patients with good function typically accrued the lowest costs, whether in an institution (17 965) or not (11 032) compared with poorer function who were living in an institution (poor: 26 370; moderate: 28,121), or community (poor: 27,207; moderate: 19,350). Hospitalization and rehabilitation services were the major costs accrued at each level of function. Many patients were left requiring a substantial amount of care and the costs associated with providing institutional care has a major impact on the economic consequences of a stroke. PMID- 15804243 TI - Reorganization of language-related neuronal networks in patients with left temporal lobe epilepsy - an fMRI study. AB - To investigate the inter- and intrahemispheric reorganization of the language cortex in left temporal lobe epilepsy (TLE) with left-sided hippocampal sclerosis. A functional magnetic resonance imaging was performed on 13 right handed patients suffering from medically intractable left TLE, and in 13 sex- and age-matched healthy controls. The activation paradigm used was a silent word generation task. A language laterality index (LI) was calculated from the number of activated voxels in the right and left anterior two-thirds of the hemispheres. Significant differences between the patients and the controls were observed in the activation of the left-sided inferior frontal gyrus. Less consistent findings in this region, as well as the relative protection of Broca's area from the activation, were revealed in the patients. In addition, different patterns of activation were proven in the cerebellum and other cortical as well as subcortical brain structures within both hemispheres. Significant differences were also found in the values of the language LIs between the investigated groups: these values suggested a more bihemispheric language representation in the patients. As anticipated, lateralization of the language functions in the epileptics significantly decreased in connection with an earlier age of initial insult. Our results support the hypothesis of a significant intra- and interhemispheric functional reorganization of language-related neuronal networks in left TLE. PMID- 15804244 TI - Whipple's disease presenting as an isolated lesion of the cervical spinal cord. AB - Chronic myelitis from Whipple's disease of the spinal cord is extremely rare. The differential diagnosis includes chronic inflammatory lesions, viral or bacterial infections, and tumours of the spinal cord. Here we present a 50-year-old man with mild sensory deficits because of a large lesion of the cervical spinal cord who markedly showed improvement during probatory antibiotic therapy. PCR and jejunal biopsy were initially negative and only later confirmed the diagnosis of Whipple's disease. Clinical and neuroradiological criteria are suggested which may be of help in the early diagnosis of spinal Whipple's disease before confirmation by molecular biology or histology. PMID- 15804245 TI - Changes of seizures activity during rapid withdrawal of lamotrigine. AB - Quantitatively evaluating the rapid withdrawal effects of lamotrigine (LTG) and carbamazepine (CBZ) on seizure activity during pre-surgical evaluation in patients with pharmacoresistant complex partial epilepsy. The duration and frequency of seizure activities and electrographic seizure onset of 41 patients totally withdrawing from CBZ monotherapy (n = 20), LTG monotherapy (n = 10) and CBZ + LTG combined therapy (n = 11) were intensively studied by therapeutic intensive seizure analysis (TISA) method. Study phases ran from the baseline phase to the antiepileptic drug (AED) withdrawal phase until the AED free phase, 3 days for each phase. Seizure duration and frequency obviously increased during the withdrawal process in each group (P < 0.05). The duration of secondarily generalized clonic signs markedly increased with the tapering of each drug; tonic signs, however, only in the AED free phase (P < 0.05). The frequency of secondary tonic and clonic signs only increased in the CBZ and CBZ + LTG group. Intergroup comparisons of all variables were insignificant (P > 0.05). There was no change of ictal EEG localization during all withdrawal protocols. All patients experienced more severe seizures during the withdrawal processes. An earlier aggravation of the clonic signs than the tonic signs was observed in each group. Difference between the withdrawal effects of LTG and CBZ monotherapy and LTG + CBZ polytherapy was mainly in the frequency change of ictal signs. The withdrawal process did not influence the ictal EEG localization. This study justified the withdrawal in pre-surgical localization, rationalized precautions for possible accompanying risks, and also aroused attentions in clinical anticonvulsant trials and substitutions involving withdrawal process. PMID- 15804246 TI - Phenotypic expression of familial amyloid polyneuropathy in Brazil. AB - Familial amyloid polyneuropathy (FAP) is an inherited amyloidosis mainly associated with transthyretin Val30Met variant. Clinical heterogeneity has been reported in different populations with FAP and Va130Met variant. In order to characterize FAP expression in Brazilians and to compare its features to those reported in other cohorts, 44 Brazilian patients (27 females, median age 36 [23 53] years) with FAP and the Val30Met variant were investigated. Approximately 40% of their family members, with the exception, of parents and siblings, had FAP. Most of the patients had symptoms of peripheral neuropathy at onset. Median age at onset was 32 [20-44] years. Earlier onset was observed in males (27 [20-43] years in males vs. 33 [20-44] years in females, P = 0.02) and in patients whose parents had FAP (31 [20-44] years vs. 40 [37-43] years in patients, respectively with and without affected parents, P = 0.03). Phenotypic expression of FAP in Brazil is similar to the one reported in Portugal, characterized by high disease penetrance, early onset, particularly in males and in subjects with affected parents, and major symptoms of peripheral neuropathy. These data highlight the influence of common genetic factors, shared by both groups of patients, in disease expression. PMID- 15804247 TI - Antigenic differences between neurological and diabetic patients with anti glutamic acid decarboxylase antibodies. AB - Antibodies to glutamic acid decarboxylase (GADAb) are found in Stiff-Person syndrome, type 1 diabetes, cerebellar ataxia and other neurological disorders (such as epilepsy and myoclonus) involving the GABAergic ways. GADAb are usually detected by immunohistochemistry (IHC), radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). This study analysed the serum of 14 patients with neurological disorders who were positive by IHC for GADAb. The performance of a commercial RIA was compared with in-house immunoblotting and ELISA methods using recombinant GAD65 (rGAD65). RIA was positive in 14 of 14, immunoblotting was positive in seven of 14 and ELISA in 12 of 14. There was no correlation between the RIA result and the ELISA optical densities. Using a sodium thiocyanate chaotrope system with ELISA to determine antibody affinity, we found no significant correlation between antibody affinity and the RIA result. A consensus should be defined concerning which assay could be used as the gold standard for detecting GADAb. The most intriguing finding was that GAD antibodies from uncomplicated diabetics do not appear to recognize GAD in frozen sections from the rat cerebellum, whereas GAD antibodies from neurologically compromised diabetics do. A working proposal is therefore that type 1 diabetic patients with unusual neurological symptoms should be tested for GADAb both by RIA and IHC. PMID- 15804248 TI - Creatine has no beneficial effect on skeletal muscle energy metabolism in patients with single mitochondrial DNA deletions: a placebo-controlled, double blind 31P-MRS crossover study. AB - The purpose of our randomized, double-blind, placebo-controlled crossover study in 15 patients with chronic progressive external ophthalmoplegia (CPEO) or Kearns Sayre syndrome (KSS) because of single large-scale mitochondrial (mt) DNA deletions was to determine whether oral creatine (Cr) monohydrate can improve skeletal muscle energy metabolism in vivo. Each treatment phase with Cr in a dosage of 150 mg/kg body weight/day or placebo lasted 6 weeks. The effect of Cr was estimated by phosphorus-31 magnetic resonance spectroscopy ((31)P-MRS), clinical and laboratory tests. (31)P-MRS analysis prior to treatment showed clear evidence of severe mitochondrial dysfunction. However, there were no relevant changes in (31)P-MRS parameters under Cr. In particular, phosphocreatine (PCr)/ATP at rest did not increase, and there was no facilitation of post exercise PCr recovery. Clinical scores and laboratory tests did not alter significantly under Cr, which was tolerated without major side-effects in all patients. Cr supplementation did not improve skeletal muscle oxidative phosphorylation in our series of patients. However, one explanation for our negative findings may be the short study duration or the limited number of patients included. PMID- 15804249 TI - Spontaneous intracerebral hemorrhage in young adults. AB - Few studies have addressed intracerebral hemorrhage (ICH) in younger adults. We studied spontaneous ICH in adults or =4 wk. Based on oral doses, subjects were assigned clinically to bi-weekly intramuscular injections of 25-75 mg of long-acting risperidone for up to 50 wk. Clinical assessments at regular intervals included the Positive and Negative Syndrome Scale (PANSS), Clinical Global Impressions (CGI) scale, adverse event reports, and the Extrapyramidal Symptom Rating Scale (ESRS). PANSS total scores improved overall from a moderate baseline score of 64.5+/-17.7 to 58.8+/-19.9 at end-point (p<0.001), by> or =20% in 50% of patients, with greatest improvement in negative symptoms. Prevalence of favourable CGI - Severity ratings increased by 2.4-fold (p<0.0001). Ratings of extrapyramidal symptoms also improved [e.g. physician-rated parkinsonism scores decreased by 20% (p<0.0001)]. Tissue reactions and other adverse effects of repeated intramuscular injections were rare and mild. Psychotic patients considered stable but symptomatic with oral risperidone treatment showed further improvements in symptom ratings and extrapyramidal dysfunction during a year of bi-weekly injections of long-acting risperidone. PMID- 15804373 TI - Evidence-based pharmacotherapy of panic disorder. AB - This paper reviews the literature on the pharmacotherapy of panic disorder, in order to address the questions (1) what is the first-line pharmacotherapy of choice for panic disorder?, (2) for how long should maintenance pharmacotherapy be continued, and (3) what is the optimal approach to the treatment-refractory patient with panic disorder. A MEDLINE search (1966-2003) was undertaken to collate randomized controlled trials of pharmacotherapy in panic disorder. A review of the evidence indicates that SSRIs are currently the first line agent of choice in panic disorder, and that pharmacotherapy should be continued for at least 1 year. There has been relatively little research on the pharmacotherapy of treatment-refractory panic disorder, and this area requires future attention. PMID- 15804374 TI - Boophilus microplus: the pattern of bovine immunoglobulin isotype responses to high and low tick infestations. AB - Cattle present variable levels of resistance to ticks and the immune correlates of these heritable phenotypes must be known in order to develop effective vaccines. The antibody responses to tick salivary antigens were examined in cattle of tick-susceptible (Holstein) and tick-resistant (Nelore) breeds. After heavy infestations, levels of IgG1 and IgG2 antibodies decreased in Holsteins and remained the same in Nelores. Conversely, levels of IgE antibodies increased in Holsteins. Different sizes of tick burdens modulated the IgG1 antibody response in a susceptible breed (Aberdeen): levels were higher than in controls in heavily infested animals, but not in those undergoing intermediary or minimal infestations. The three experimental groups presented similar levels of IgG2 antibodies. Levels of IgE antibodies were higher only in animals undergoing intermediate infestations. These results indicate that tick infestations suppress the IgG antibody response in susceptible breeds, that IgE antibodies are not protective, and that the dose of tick saliva modulates the isotype of host antibody responses. PMID- 15804375 TI - Uronema marinum: identification and biochemical characterization of phosphatidylcholine-hydrolyzing phospholipase C. AB - Phosphatidylcholine (PC)-specific phospholipase D (PC-PLD) and phosphatidylcholine (PC)-specific phospholipase C (PC-PLC) activities have been detected in Uronema marinum. Partial purification of PC-PLC revealed that two distinct forms of PC-PLC (named as mPC-PLC and cPC-PLC) were existed in membrane and cytosol fractions. The two PC-PLC enzymes showed the preferential hydrolyzing activity for PC with specific activity of 50.4 for mPC-PLC and 28.3 pmol/min/mg for cPC-PLC, but did not hydrolyze phosphatidylinositol or phosphatidylethanolamine. However, the biochemical characteristics and physiological roles of both enzymes were somewhat different. mPC-PLC had a pH optimum in the acidic region at around, pH 6.0, and required approximately 0.4 mM Ca2+ and 2.5 mM Mg2+ for maximal activity. cPC-PLC had a pH optimum in the neutral region at around, pH 7.0, and required 1.6 mM Ca2+ and 2.5 mM Mg2+ for maximal activity. cPC-PLC, but not mPC-PLC, showed a dose-dependent inhibitory effect on the luminal-enhanced chemiluminescence (CL) responses and the viability of zymosan-stimulated phagocytes of olive flounder, indicating that cPC-PLC may contribute to the parasite evasion against the host immune response. Our results suggest that U. marinum contains PC-PLD as well as two enzymatically distinct PC PLC enzymes, and that mPC-PLC may play a role in the intercellular multiplication of U. marinum and cPC-PLC acts as a virulence factor, serving to actively disrupt the host defense mechanisms. PMID- 15804376 TI - The detection of non-RoTat 1.2 Trypanosoma evansi. AB - The majority of Trypanosoma evansi can be detected using diagnostic tests based on the variant surface glycoprotein (VSG) of Trypanosoma evansi Rode Trypanozoon antigen type (RoTat) 1.2. Exceptions are a number of T. evansi isolated in Kenya. To characterize T. evansi that are undetected by RoTat 1.2, we cloned and sequenced the VSG cDNA from T. evansi JN 2118Hu, an isolate devoid of the RoTat 1.2 VSG gene. A 273 bp DNA segment of the VSG gene was targeted in PCR amplification for the detection of non-RoTat 1.2 T. evansi. Genomic DNA samples from different trypanosomes were tested including 32 T. evansi, 10 Trypanosoma brucei, three Trypanosoma congolense, and one Trypanosoma vivax. Comparison was by PCR amplification of a 488 bp fragment of RoTat1.2 VSG gene. Results showed that the expected 273 bp amplification product was present in all five non-RoTat 1.2 T. evansi tested and was absent in all 27 RoTat 1.2-positive T. evansi tested. It was also absent in all other trypanosomes tested. The PCR test developed in this study is specific for non-RoTat 1.2 T. evansi. PMID- 15804377 TI - Flow cytometric assessment of Leishmania spp metacyclic differentiation: validation by morphological features and specific markers. AB - Characterization of infective metacyclic promastigotes of Leishmania spp can be an essential step in several experimental protocols. Metacyclic forms of all Leishmania species display a typical morphology with short, narrow cell body, and an elongated flagellum. This feature suggests that metacyclics can be distinguished from procyclic forms by non-fluorimetric flow cytometric parameters thus enabling the follow-up of their appearance and acquisition of specific properties, during metacyclogenesis in in vitro cultures. Here we describe the flow cytometric parameters of stage-specific promastigotes of Leishmania major, Leishmania donovani, Leishmania amazonensis, and Leishmania braziliensis. Our findings were validated by optical microscopy morphology and specific procyclic labeling with FITC-peanut agglutinin. Furthermore, we show that parasite's distribution in the plot during differentiation in culture is not species specific and that the parasites displaying low forward-angle light scatter (FSC(low)) are three times more infective than the FSC(high) ones. The method here described can be applied to the identification of metacyclics of different Leishmania spp within the whole stationary population. PMID- 15804378 TI - Neospora caninum: quantification of DNA in the blood of naturally infected aborted and pregnant cows using real-time PCR. AB - This study quantified Neospora caninum DNA in the blood and brain of pregnant and aborted heifers by monitoring PCR product formation in real-time using SYBR Green I, a double-stranded DNA-binding dye. Primers were designed to amplify a 188 bp product specific to N. caninum from the Nc-5 gene fragment of N. caninum. Similarly, a 71 bp product was amplified from the 28S rRNA gene of bovine genomic DNA that served as a control. Agarose gel electrophoresis and analysis of the melting curve for PCR products showed that both primer pairs were specific to their targets. Standard curves were generated for both bovine and N. caninum genomic DNA, and were used to compute the relative concentration of parasite to bovine DNA in the test samples. The concentration of N. caninum DNA in 1 ng of bovine genomic DNA obtained from blood ranged between 0.097 ng at the 1st week of the observation and 0 ng at the 15th week in aborted cows. In pregnant cows, the values ranged between 0.080 ng at the 1st week and 0.155 ng at the 15th week of observation. There was a sustained decrease of DNA concentration in the aborted group after abortion and an increase in DNA concentration in the pregnant group. Comparison of parasite DNA in blood and brain of infected heifers showed a higher DNA concentration in brain than in blood. This study shows that N. caninum DNA can be quantified to obtain the relative concentration of parasite DNA to host genomic DNA in blood. This technique allows testing of a large number of samples at one time and can be done without the need for slaughter of tested animals. PMID- 15804379 TI - The larval feeding inhibition assay for the diagnosis of nematode anthelmintic resistance. AB - A larval feeding assay for detection of nematode anthelmintic resistance to macrocyclic lactones and imidazothiazoles is described. The estimated concentration of anthelmintic required to inhibit larval feeding in 50% of L1's (IC50) that were resistant to either macrocyclic lactones or imidazothiazoles were significantly higher (P < or = 0.05) than those of susceptible isolates. Some variations in IC50 values were observed during the patent period of infection in all strains, although the pattern of the IC50 followed the same course. IC50 values varied in larvae developing from eggs shed throughout the patent period, with low values in the earliest larvae followed by higher values as the infection progressed, before decreasing at 70-90 days post-infection, although the low values of the first part of the patent period were not reached. However, the IC50 differences between all resistant and susceptible strains were significant throughout the whole patent period for ivermectin and levamisole. These results suggest that this technique may provide an alternative in vitro to detect anthelmintic resistance. PMID- 15804380 TI - Giardia lamblia: evaluation of the in vitro effects of nocodazole and colchicine on trophozoites. AB - Giardia lamblia is the most commonly detected parasite in the intestinal tract of humans and other mammals causing giardiasis. Giardia presents several cytoskeletal structures with microtubules as major components such as the ventral adhesive disk, eight flagella axonemes, the median body and funis. Many drugs have already been tested as antigiardial agents, such as albendazole and mebendazole, which act by specifically inhibiting tubulin polymerization and hence microtubule assembly. In the present work, we used the microtubule inhibitors nocodazole and colchicine in order to investigate their direct and indirect effects on Giardia ultrastructure and attachment to the glass surface, respectively. Axenically grown G. lamblia trophozoites were treated with nocodazole or colchicine for different time intervals and analyzed by light and electron microscopy. It was observed that trophozoites became completely misshapen, detached from the glass surface and failed to complete cell division. The main alterations observed included disc fragmentation, presence of large vacuoles, and appearance of electrondense deposits made of tubulin. The cytokinesis was blocked, but not the karyokinesis, and membrane blebs were observed. These findings show that Giardia behavior and cytoskeleton are clearly affected by the commonly used microtubule targetting agents colchicine and nozodazole. PMID- 15804381 TI - Plasmodium falciparum: evaluation of a quantitative nucleic acid sequence-based amplification assay to predict the outcome of sulfadoxine-pyrimethamine treatment of uncomplicated malaria. AB - A quantitative nucleic acid sequence-based amplification (QT-NASBA) assay was employed to predict retrospectively the outcome of sulfadoxine-pyrimethamine (SP) treatment of uncomplicated malaria in children aged <6 years in an endemic region. Blood samples were collected at initial diagnosis and during follow-up. Mutation-specific nested PCR methods to analyse DHFR (Arg-59) and DHPS (Glu-540) mutations that are associated with SP drug resistance were applied. Parasite genotyping was performed to distinguish between re-infection and recrudescence. Eighty-six patients were recruited of which 66 were available for follow-up. Nine children were classified as early treatment failure, 13 cases were classified as late clinical failure, 32 as late parasitological failure, and only 12 children had an adequate clinical and parasitological response. DHFR and DHPS mutations conferring SP resistance were abundant in the Plasmodium population. Blood samples obtained 7 days after treatment were used to predict retrospectively the outcome of SP treatment. QT-NASBA was able to give a correct prediction of treatment outcome in 85.7% of the cases. Positive predictive value (PPV) of QT NASBA case was 95% (95% confidence interval = 88.3-100) and negative predictive value (NPV) was 63% (95% CI = 39.5-86.5). In contrast, microscopy correctly predicted outcome in only 37.5% of the cases. PPV of microscopy was 100% (95% CI = 73.9-100) and the NPV was 25.5% (95% CI = 13.0-38.0). The analysis of a day 7 blood sample with QT-NASBA allows for the prediction of late clinical or parasitological treatment failure in the majority of the cases analysed in the present study. PMID- 15804383 TI - Echinococcus granulosus: pre-culture of protoscoleces in vitro significantly increases development and viability of secondary hydatid cysts in mice. AB - We describe a method for obtaining improved secondary infections of Echinococcus granulosus that involves culturing protoscolex larvae in vitro prior to inoculation into mice. This approach provides a far superior method for obtaining secondary echinococcosis infections in mice compared with the traditional method of direct inoculation of protoscoleces (PSC) where the majority of parasites are killed by the host. We obtained a high rate of recovery both in terms of secondary cyst numbers and their viability. After 50 weeks post-infection (p.i.), brood capsules were formed and the first PSC developed in each of the capsules. After 56 weeks p.i., the fastest developing brood capsule contained four PSC. The approach will prove valuable for investigating parasite development and the host parasite interaction in secondary echinococcosis. PMID- 15804382 TI - Plasmodium yoelii: a differential fluorescent technique using Acridine Orange to identify infected erythrocytes and reticulocytes in Duffy knockout mouse. AB - Both human malarial parasite Plasmodium vivax and mouse malaria parasite Plasmodium yoelii use Duffy protein as the receptor for invasion and they preferentially invade reticulocytes. Recently, it has been shown that P. yoelii invades mouse reticulocytes by a Duffy independent pathway. Parasite invasion is generally visualized by time consuming staining procedures with dyes like Giemsa or Wright-Giemsa. Fluorochromatic dye like Acridine Orange has been used for instantaneous detection of parasites in RBCs. Acridine Orange binds to both DNA and RNA but with different emission spectra; and the binding can be distinguished with a fluorescent microscope using a green or a red filter, respectively. We have used this differential emission of Acridine Orange to determine P. yoelii invasion into erythrocytes and reticulocytes of Duffy positive and Duffy knockout mice. Moreover, we show that this method can be used to determine the maturity of reticulocytes in the peripheral blood of anemic mice. PMID- 15804384 TI - Increased volume of the pigmented neurons in the locus coeruleus of schizophrenic subjects: a stereological study. AB - The locus coeruleus is the largest cluster of noradrenaline-producing neurons in the brain and has been involved in regulating attention. The neurotransmitter system contributes to the initiation and maintenance of forebrain activity as well as modulation of the collection and processing of sensory information. This makes locus coeruleus a target of interest in the study of possible structural changes in the brains of subjects with chronic schizophrenia. Uniform sampling and optical disectors were used for estimation of total neuron numbers and the rotator principle for estimation of mean cell volume. This study estimated the bilateral total number of pigmented neurons in the locus coeruleus of schizophrenic and control subjects and found no difference between the two; schizophrenic subjects have 37,400 (coefficient of variation=CV=SD/mean=0.28), control subjects have 35,500 (CV=0.19), p=0.64. The average volume of the cell perikaryon of pigmented neurons was measured in both groups and showed a significant larger cell volume in the locus coeruleus of schizophrenic compared to control subjects; mean locus coeruleus cell volume was 58,400 microm(3) (CV=0.31) in schizophrenic subjects, which is 55% larger than the 37,600 microm(3) (CV=0.33) found in control subjects, a difference of 20,800 microm(3) (p=0.009). The fixation time was significantly longer for the schizophrenic brains with an average of 169 months compared to 88 months for the control brains. No relation was found between total numbers or geometric mean volume of the pigmented cells and fixation time. PMID- 15804385 TI - Anatomical MRI study of corpus callosum in unipolar depression. AB - Previous studies have suggested abnormal cerebral lateralization in major depressive disorder (MDD). Few controlled MRI studies have investigated the corpus callosum (CC), the largest commissura connecting the two cerebral hemispheres, in MDD. This study investigated anatomical abnormalities in the CC and its subdivisions in MDD patients. Twenty-two unmedicated MDD patients and 39 healthy subjects underwent brain magnetic resonance imaging (MRI). Measurements of the CC and its sub-regions were performed with a semi-automated software (NIH Image, version 1.62). ANCOVA with age, gender, and intra-cranial volume (ICV) as covariates showed no significant differences in CC measurements between patients and controls (df=1,56; p>0.05). However, patients with familial MDD had a significantly larger middle genu area (F(1,45)=4.252; p=0.045) compared to healthy controls, and significantly larger middle genu (F(1,13)=5.366; p=0.037), anterior splenium (F(1,13)=6.27; p=0.026), and middle splenium areas (F(1,13)=4.706; p=0.049) compared to patients with non-familial MDD. Although preliminary, our findings suggest that anatomical abnormalities in CC may be restricted to patients with familial MDD, with possible enlargement of CC in this particular sub-group. The possible role of callosal abnormalities in the pathogenesis of mood disorders should be further examined. PMID- 15804386 TI - Volumetric MRI measurement of caudate nuclei in antipsychotic-naive patients suffering from a first episode of psychosis. AB - Magnetic resonance imaging (MRI) studies measuring basal ganglia volumes in first episode patients suggest that treatment with typical neuroleptics leads to alteration in these brain structures. However, caudate nuclei volumes (CNV) of untreated first-episode patients may even be smaller than in healthy controls. We investigated whether CNV of newly diagnosed neuroleptic-naive psychotic patients differ as compared to an age- and sex-matched healthy control group to detect possible treatment effects early in the course of this illness. Magnetic resonance images were acquired in 37 un-medicated psychotic patients and 37 healthy controls. Ten of the patients were re-examined after 12 weeks of treatment with the second generation antipsychotic quetiapine. Regions of interest (ROI) delineating the caudate nuclei bilaterally were drawn manually using Brain Image software. The neuroleptic-naive patients showed a mean CNV of 8.40 cc (SD=1.01) and the controls of 8.55 cc (SD=1.16). There was no significant difference between groups (F=.600; P=.441). In contrast to previous studies in patients treated with typical neuroleptics, this cross-sectional MRI study did not find significant differences in CNV of neuroleptic-naive first-episode patients compared to healthy controls. PMID- 15804387 TI - Association of the serotonin transporter gene polymorphism with Korean male alcoholics. AB - This study investigated the association between the serotonin transporter polymorphism (5-HTTLPR) and alcoholism in the Korean population. In addition, in order to reduce the clinical heterogeneity, sub-analysis was carried out according to some clinical variables such as a family history of alcoholism, aggressive/violent behavior and the age of onset of alcoholism. One hundred and forty-five patients meeting the DSM-VI criteria for alcohol dependence and 201 healthy controls were examined. Genotyping was performed using a polymerase chain reaction (PCR)-based method. The frequency of the L-allele of 5-HTTLPR was significantly higher in the alcohol dependent patients than in the normal controls (chi(2)=19.11, df=1, p<0.001). Furthermore, there was a significant difference in the allelic distribution between the subgroups defined by a family history of alcoholism (chi(2)=4.005, df=1, p=0.045). This study suggests a putative role of the 5-HTTLPR for alcoholism in the Korean population. However, a replication study with larger different ethnic samples and a refinement of the subtype of alcoholism is needed. PMID- 15804389 TI - Cognitive dysfunctions in medicated and unmedicated patients with recent-onset schizophrenia. AB - Schizophrenia is associated with impairments in many cognitive domains on which the influence of antipsychotics, whether conventional or atypical, remains unclear. We conducted a study of recent-onset schizophrenic patients (DSM IV) that included unmedicated (n=19), and medicated (n=19) patients matched for age and IQ. Both groups of patients had comparably low extra-pyramidal symptoms (EPS). Cognitive tasks included attentional tasks (alertness and divided attention tests), a working memory task (a verbal n-back test) and the Wisconsin Card Sorting Test (WCST). After adjustment for the Total PANSS score, we found no significant difference between the two groups of patients in any of the cognitive tasks. When compared to a group of healthy controls (n=20) matched for IQ level, unmedicated patients performed significantly worse in all cognitive tasks, with significantly longer reaction times for alertness, divided attention and working memory. These results confirm the presence of cognitive impairments in attentional and executive functions in recent-onset patients whether or not they are medicated. There was no evidence that either conventional or atypical antipsychotics had an influence on patients when EPS were excluded. Altogether, our results further support the idea that cognitive deficits in schizophrenia are enduring features per se and cannot be considered as secondary to psychiatric symptoms or to the adverse effects of medication. In addition our results suggest that antipsychotics do not have a major effect on these impairments. PMID- 15804388 TI - Prevalence and predictors of pathological gambling: results from the St. Louis personality, health and lifestyle (SLPHL) study. AB - OBJECTIVES: We report the prevalence of and risk and protective factors for DSM IV sub-threshold gambling (1-4 criteria) and pathological gambling disorder (PGD; 5-10 criteria) in a non-clinical household sample of St. Louis area gamblers. METHODS: Of the 7689 individuals contacted via Random Digit Dialing, 3292 were screened eligible. Of these, 1142 from households in 6 contiguous regions in Missouri and Illinois consented to participate and were mailed a St. Louis Area Personality, Health, and Lifestyle (SLPHL) Survey. RESULTS: Post-stratification weighted data (n=913) indicate lifetime prevalence rates of 12.4% sub-threshold and 2.5% PGD (conditional prevalence=21.5% and 4.3% respectively). Risk and protective factors for gambling severity varied in the sample. CONCLUSIONS: Targeted prevention messages are warranted specifically for gamblers of varying risk for PGD. PMID- 15804390 TI - Obsessive-compulsive disorder (OCD) with schizotypy vs. schizophrenia with OCD: diagnostic dilemmas and therapeutic implications. AB - Although schizophrenia and obsessive-compulsive disorder (OCD) are distinct diagnostic entities, there are substantial areas of overlap between the two disorders in clinical characteristics, affected brain areas and pharmacotherapy. Though OCD patients apparently do not have increased risk for developing schizophrenia, schizotypal personality disorder has consistently been found in OCD patients. Compelling evidence also points to an increased rate of OCD in schizophrenia patients. Accurate diagnosis of both disorders in their "pure" and overlapping forms is necessary in order to evaluate etiological mechanisms underlying schizophrenia and OCD, and to provide adequate treatment and prognosis. In this review, we address some aspects of the current status of research pertinent to the OCD-schizophrenia interface and suggest further steps towards the clinical and etiological identification of homogeneous subgroups on the putative OCD-schizophrenia axis. PMID- 15804391 TI - Anxiety disorders in outpatients with schizophrenia: prevalence and impact on the subjective quality of life. AB - OBJECTIVE: To describe the prevalence of comorbid lifetime anxiety disorders in outpatients with schizophrenia and to compare the subjective quality of life of patients with and without comorbid anxiety disorders. METHODS: Fifty-three outpatients were recruited. They were interviewed with the Anxiety Disorders section of the SCID for DSM-IV. Quality of life was assessed with the Sheehan disability scale (SDS). RESULTS: Specific prevalences of anxiety comorbidity were: social phobia (17%), OCD (15.1%), GAD (9.4%), anxiety disorder NOS (7.5%), panic disorder (5.7%), specific phobia (5.7%), PTSD (3.8%), and agoraphobia (1.9%). Schizophrenic patients with comorbid anxiety disorder (41.5%) showed significantly higher scores in global scale (p=0.005), work subscale (p=0.007), and social life subscale (p=0.003) of the SDS than their counterparts without comorbid conditions. CONCLUSIONS: Anxiety disorders may impose an additional burden to patients with schizophrenia, resulting in further decline in their subjective quality of life. PMID- 15804392 TI - Stability of DSM-IV criterion symptoms for major depressive disorder. AB - Given the chronic and recurrent nature of major depressive disorder (MDD), it is important to understand whether specific symptoms are stable over time or vary over the course of the disorder. This is the first longitudinal investigation examining the stability of the nine criterion symptoms of depression, as specified in the DSM-IV, among diagnosed depressed adults who were not recovered at follow-up. In this study, participants were assessed twice, ten months apart, with the structured clinical interview for DSM-IV, and stability of the nine criterion symptoms of MDD was examined. Findings indicate strong stability in individuals' symptom profiles. Among individuals who were clinically depressed at both assessments, there were no statistically significant fluctuations in specific symptoms endorsed. Changes in symptom endorsement among individuals who no longer met diagnostic criteria for MDD at Time 2 were attributable to reduced severity (i.e., number of symptoms) rather than to inconsistency of symptom endorsement. These results indicate that depressed individuals experience essentially the same pattern of specific symptoms over the course of a year. Variation in clinical course is likely to be attributable more to fluctuations in overall severity than to changes in specific symptoms of depression. PMID- 15804393 TI - High prevalence of personality disorders among healthy volunteers for research: implications for control group bias. AB - Individuals who volunteer as control subjects for clinical studies are regularly screened for Axis I diagnoses, but seldom screened for Axis II disorders. This study examined the relative rates of Axis II diagnoses among 341 volunteers passing an initial telephone screen for entry into biological research studies. Axis I and II diagnoses by DSM-IV were assigned by best estimate after structured clinical interview, and subjects were categorized into one of three groups based on their diagnostic profiles: (1) volunteers without lifetime Axis I or II diagnoses ("healthy controls"), (2) personality-disordered volunteers without any history of Axis I pathology, and (3) personality-disordered volunteers with past (but not current) Axis I pathology. The results revealed a high prevalence of personality disorders (44.4%) among these volunteers. Several clinically relevant self-report inventories were used to demonstrate important characterological differences between the three comparison groups. Although inventory results demonstrated multiple differences between all three groups, most scales revealed differences between healthy controls and the two personality-disordered groups (with or without lifetime Axis I diagnoses), suggesting that most of the variance was accounted for by the presence or absence of an Axis II disorder, not a past Axis I disorder. These results suggest that personality-disordered volunteers may bias a control group due to the infrequent screening for Axis II disorders among volunteers for medical and psychiatric research. Implications are discussed for routine Axis II screening of volunteers for research with specific diagnostic instruments. PMID- 15804394 TI - Elevated pain threshold in eating disorders: physiological and psychological factors. AB - Several studies have found decreased pain sensitivity in patients with eating disorders but it is unclear what physiological and psychological factors are associated with this abnormality. In the present investigation, thermal pain threshold latency, somatoform dissociation, body image disturbance and physiological indices of autonomic neural system activity were measured in 39 female patients with eating disorders (21 with anorexia nervosa and 18 with bulimia nervosa) and 17 healthy women. Pain threshold was elevated in patients with eating disorders, especially those with binge-purging symptomatology. A regression analysis indicated that increased pain threshold is moderately associated with decreased peripheral skin temperature and weakly associated with lack of familiarity with one's own body. However, the between group differences in pain perception remained significant after controlling for peripheral skin temperature. Hence, elevation of pain threshold in eating disorders is a replicable finding, which is not explicable by peripheral indices of autonomic system activity or by somatoform dissociation. In future research it may be evaluated as a potential marker of broader phenotype of decreased interoceptive awareness, which may be associated with vulnerability to the development of eating disorders. PMID- 15804395 TI - Clinicians' diagnosis of a case with anger problems. AB - Psychiatrists and psychologists responded to case vignettes to assess the prevalence, severity, and diagnostic confidence clinicians had concerning treating anger disordered clients compared with clients with generalized anxiety disorder. Five hundred and forty-two clinicians (a response rate of 30%) assessed one of two matched case histories by mail. One described generalized anxiety disorder (GAD) and the other a case of anger disorder (AD). Cases were identical except for thoughts and affect relevant to the disorders. Both male and female versions were used. More than 95% of the participants viewed the cases they received as pathological. The disorders were rated as equally common. The clinicians reported treating equal numbers of patients with similar anger or anxiety symptoms in the past year. Although the case histories were alike in length and detail, AD participants rated their case as less complete and had lower confidence in their diagnoses. The diagnostic consensus was high for GAD clinicians, but low for AD. Forty-three percent of participants selected an Axis II diagnosis for AD, compared with 3% for GAD. Clinicians appeared to encounter patients with chronic anger about as frequently as they see GAD, but they displayed diagnostic confusion and bias toward personality disorder diagnoses when presented with the anger symptoms. The findings support the development of a diagnostic category for primary anger. The wide dispersion of diagnoses for anger underscores the need for focused differential assessment. PMID- 15804396 TI - Reduced free tri-iodothyronine serum levels are lower in pathological male gamblers than in healthy male controls. PMID- 15804397 TI - Postnatal development of limb motor innervation in the opossum Monodelphis domestica: immunohistochemical localization of acetylcholine. AB - The development of limb motor innervation was studied in the opossum Monodelphis domestica, a marsupial born with immature mobile forelimbs and immobile hindlimbs. Choline acetyltransferase (ChAT), the synthesis enzyme of acetylcholine, was evidenced on sections of the spinal enlargements, and the protein that transports acetylcholine (VAChT) on limb sections. In newborn, ChAT immunolabeling occurred in small, undifferentiated neurons of the ventral horn, presumably motoneurons, and intermediate and dorsal gray matter, and in the presumptive white matter, all less abundant at lumbosacral than brachial levels. Scant immunolabeling for VAChT marked small terminal-looking profiles, presumably growth cones or immature neuromuscular junctions, decreasing proximodistally in each limb and being less abundant in hindlimbs than forelimbs; it was absent distally in the foot where no muscle tissue was formed. ChAT labeling disappeared from the white matter within 1 week while cholinergic neurons increased in number and size. Motoneurons segregated in a medial and lateral group by 4-5 weeks. VAChT-labeled profiles increased in number and size and they flattened along a proximodistal gradient within each limb, but later in the hindlimbs than in the forelimbs. Labeling appeared in distal foot muscle at 1 week. The density, size, and shape of terminals became comparable in all segments of a given limb by 3-4 weeks. Their number and size increased, and by 8 weeks, they clustered in 3 or 4 along muscle fibers. Thus, limb motor innervation develops largely postnatally in the opossum, along rostrocaudal and proximodistal gradients. Its timecourse is compared to the development of motor behaviors. PMID- 15804398 TI - Mutation at the Lmx1a locus provokes aberrant brain development in the rat. AB - A rat short-tail mutation with neurological defects (named queue courte, qc) was discovered. Histopathology in adult qc/qc rats revealed hypoplasia of the cerebellum and hippocampus, maldevelopment of the choroid plexus and corpus callosum. These abnormalities are strongly reminiscent of the phenotypic abnormalities found in the shaker short-tail or dreher (dr) mouse mutation at the LIM homeobox transcription factor 1 alpha locus (Lmx1a). The qc mutation is an autosomal recessive and has been mapped to the dr homologous region on rat chromosome 13, and Northern blot analysis demonstrated no expression of Lmx1a in qc/qc rats. Narrowing and distortion of the ventricles were observed from embryonic day 17 (E17) in qc/qc rats. From E17, fusion of the opposing neuroepithelium and formation of neuroepithelial rosettes were also found. Arrangements of neuroepithelial cells were disturbed and processes of radial glia were disoriented in the fused lesions. Neuronal migration analysis using BrdU immunohistochemistry revealed defective migration from the neuroepithelium toward the neocortex and mesencephalon in qc/qc rats. These findings suggest that the qc mutation is involved in development of the ventricular system and dorsal migration of neurons. PMID- 15804399 TI - Dimorphic expression of testosterone metabolizing enzymes in the hypothalamic area of developing rats. AB - Androgen transformation into estrogens through the aromatase enzyme, occurring in the rat hypothalamus during fetal life, leads to male-specific sexual differentiation of brain. Aromatase shows a peak of expression and activity in a limited period during late gestation; however, the possible dimorphism in its expression during embryogenesis is unclear. One of the mechanisms controlling tissue-specific aromatase expression might be the formation of transcript variants, that differ in the 5'-untranslated regions (5'-UTR). Exon If is the major 5'-UTR used in rodent hypothalamic-preoptic area, with low amounts of other variants encoded by different exons I also present. Another enzymatic conversion, possibly involved in brain differentiation, is the 5 alpha-reduction of Testosterone to DHT, catalyzed by two 5 alpha-reductases (5 alpha-R type1 and 2). Aim of the present study is to evaluate, in parallel, by semiquantitative RT-PCR, the dimorphic profile of the three enzymes and the pattern of the brain-specific aromatase expression in male and female rats from gestation-day 16 to postnatal day 5 (or 15 only for 5 alpha-R1). It has been observed that, in both sexes, 5 alpha-R1 is significantly higher around birth than prenatally, and that 5 alpha R2 expression appears to be higher in males than in females, particularly just after birth. Moreover, aromatase has two expression peaks, that are male specific, before and after birth; only exon If is used in males, while different transcripts might be present in females postnatally. It is concluded that rodent brain sexual differentiation probably involves the activation of both 5 alpha-R2 and aromatase enzymes in a sex- and time-specific pattern. PMID- 15804400 TI - Axonal projections from the hypothalamus to the pituitary intermediate lobe in rats during ontogenesis: DiI tracing study. AB - This study has determined ontogenetic schedule of axonal arrival from the hypothalamus in the pituitary intermediate lobe (IL) in rats using 1,1' dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) as a retrograde tracer. The brains with attached pituitaries were dissected in rats from the 20th embryonic day (E20) to the 20th postnatal day (P20). Anterior lobe was mechanically detached from the IL, material fixed in paraformaldehyde, and DiI crystals were applied on the IL laying on the posterior lobe (PL). The labeling of IL + PL resulted in staining of hypothalamic magnocellular neurons, which send their axons to the PL, and hypothalamic parvocellular neurons contributing to the innervation of the IL. Therefore, the magnocellular neurons were not taken into account when identifying the neurons projecting axons to the IL. Rare fluorescent neurons projecting their axons to the IL were detected as early as on E20 in the ventral part of the periventricular nucleus (Pe) and in the rostral part of the arcuate nucleus. Few DiI-labeled neurons were seen in Pe from P1 to P3. At P5, the fluorescent neurons were accumulated giving rise to the prominent cluster in the Pe, which was enlarged on later stages and occupied all the Pe. In addition to the Pe, fluorescent neurons first appeared in the retrochiasmatic region and around the ventromedial nucleus in young rats. Thus, the axons of hypothalamic neurons of the Pe and mediobasal hypothalamus first arrive in the IL in rats at the end of intrauterine development, although the principal innervation of the IL is the postnatal event. PMID- 15804401 TI - Neutrophil-specific chemokines are produced by astrocytic cells but not by neuronal cells. AB - BACKGROUND: Neutrophils have a central role in the inflammatory conditions of the central nervous system (CNS). ELR chemokines direct neutrophil migration, but the source of chemokines in the CNS is unclear. We quantified chemokine production using cell-line models of astrocytic and neuronal cells, specifically NT2.N cells, a human line with characteristics of immature neurons, and NT2.A cells, a line with characteristics of astrocytes. OBJECTIVE: In NT2.N and NT2.A cells, and their parent cell line NT2, we sought to: (1) quantify ELR chemokines, (2) determine receptor (CXCR-1 and CXCR-2) expression, and (3) measure the function of the chemokines generated from these cells. DESIGN/METHODS: NT2 cells were differentiated into NT2.N cells and NT2.A cells with all trans retinoic acid and mitosis inhibitors. Chemokine concentrations in culture supernatants were determined by ELISA. Immunofluorescence was used to detect CXCR-1 and CXCR-2. RT PCR was used to determine chemokine and chemokine receptor mRNA. Chemotaxis assays were used to assess function. RESULTS: ELR chemokines were not detected in supernatants of NT2 or NT2.N cells, although mRNA for GRO-gamma/CXCL3 was found in both. In contrast, in NT2.A cells, mRNA and protein were present for GCP 2/CXCL6, GRO-alpha/CXCL1, GRO-gamma/CXCL3, and IL-8/CXCL8. CXCR-1 and CXCR-2 were expressed on NT2, NT2.N, and NT2.A cells detected by immunofluorescent staining and RT-PCR. Supernatants of NT2.A cells resulted in neutrophil chemotactic function of 30.5 +/- 3.9%, greater than NT2 cells (12.3 +/- 1.6%, mean +/- SEM, P < 0.01). CONCLUSIONS: We speculate that astrocytes are a source of ELR chemokines in the human CNS and that neurons and astrocytes can respond to those chemokines. PMID- 15804402 TI - Effects of prenatal alcohol exposure on the development of the vibrissal somatosensory cortical barrel network. AB - We have previously shown that the serotonin (5-HT) and its thalamocortical afferents are compromised by prenatal alcohol exposure (PAE). The development of the sensory cortical barrels is regulated by 5-HT-rich thalamocortical afferents. Therefore, it is hypothesized that PAE will deleteriously affect the postnatal development of the cortical barrel formations. On embryonic day (E)7, C57BL/6 mice were grouped into: Alcohol (Alc), Pair-fed (PF), or Chow, and maintained on diet until E18. On postnatal day 7, cortices were stained with 5-HT for thalamocortical fibers, and a NeuN for identification of mature neurons. The area of the posterior medial barrel subfield (PMBSF), was measured as well as the number of NeuN+ neurons within the barrel patches. Though brain weight and brain volume were similar among the three groups, a significant reduction was seen in total area of the PMBSF, and in the average individual barrel area in the Alc group as compared to Chow. Furthermore, the volumes of the B, but not C row barrels were significantly reduced. Barrels were found missing in layer IV, specifically in the posterior aspects of the A, B, and straddler row in the Alc group. Cell counts demonstrated a nearly 50% reduction in NeuN+ neuron number in both rows. This reduction in size of the PMBSF and fewer neurons within these sensory barreloids may underlie a change in the development of the discriminatory sensitivity of the whiskers and serves as an excellent model for the study of a compromised sensory modality following PAE. PMID- 15804403 TI - Maturation of the hypothalamic arcuate agouti-related protein system during postnatal development in the mouse. AB - The hypothalamic arcuate nucleus (Arc) and its neurons expressing agouti-related protein (AgRP) are key components of the forebrain circuitry involved in long term regulation of energy homeostasis, including conveying leptin signaling to other hypothalamic and extrahypothalamic regions. In the present work, we investigated the postnatal development (P0, P5, P10, P15, and P21) of this system (AgRP transcript and peptide) in the mouse brain using in situ hybridization and immunohistochemistry. At all stages, AgRP mRNA expression was detected exclusively in the Arc. At P0, AgRP mRNA levels were low, and only a few AgRP immunoreactive fibers were present reaching, rostrally, the bed nucleus of the stria terminalis and, caudally, the dorsal raphe nucleus. During the following period (P5-P21), the levels of AgRP mRNA gradually increased in the Arc along with a parallel increase in the AgRP fiber density in the hypothalamic regions responsible for control of appetite, including the paraventricular nucleus, as well as in extrahypothalamic regions, including locus coeruleus. These data provide evidence that, in the mouse, the maturation of the AgRP Arc system occurs mainly during the first three postnatal weeks. Together with the existing data on the physiology of appetite and body weight, our data suggest that the first three postnatal weeks in the mouse represents a critical period for the formation of brain mechanisms underlying appetite control via peripheral hormones. PMID- 15804404 TI - Expression of glutamate transporter subtypes during normal human corticogenesis and type II lissencephaly. AB - Glutamate transporters are thought to have an important role in central nervous system (CNS) development. We investigated the expression of the sodium-dependent high-affinity glutamate transporters EAAT1, EAAT2, and EAAT3 in 11 human autopsied cases without neurological disorders and in four cases with type II lissencephaly including Walker Warburg's syndrome (WWS) and Fukuyama-type congenital muscular dystrophy (FCMD), both of which are classified as migration disorders of the human brain. Expression of glutamate transporter subtypes was differentially regulated during normal human corticogenesis. Although EAAT1 and EAAT2 were mainly localized to the cortical astrocytes in the postnatal brain, EAAT1 was enriched in the proliferative zones and radial glia from 13 gestational weeks (GW) to 20 GW. EAAT2 was abundant in the intermediate zone until 23 GW, and transiently expressed in the radial fibers of the transitional form of radial glia into mature astrocytes as well as partly in the corticofugal axonal bundles. EAAT3 immunoreactivity was robust in the apical dendrites of the pyramidal neurons in the marginal zone and cortical plate during corticogenesis, and decreased postnatally. In the individuals with type II lissencephaly, glutamate transporters were expressed in the extrusion of neuroglial tissue. Bundles of EAAT2-immunoreactive radial fibers were prominent in the specimens at 20 GW. Thus, glutamate transporters are differentially regulated during normal and impaired corticogenesis. Altered glutamate transporter expression in type II lissencephaly suggests that glutamate metabolism is involved in the formation of the normal cortex and contributes to the disorganized cortex seen in migration disorders. PMID- 15804405 TI - Lack of NMDA receptor subunit exchange alters Purkinje cell dendritic morphology in cerebellar slice cultures. AB - Early postnatal developmental changes in N-methyl-d-aspartate (NMDA) receptor (NR) subunits regulate cerebellar granule cell maturation and potentially Purkinje cell development. We therefore investigated Purkinje cell morphology in slice cultures from mice with genetic subunit exchange from NR2C to NR2B (NR2C 2B). NR2C-2B Purkinje cells after 12 days in vitro showed a significantly impaired dendritic arbour complexity with reduced branching density as compared to wild-type cells, a phenotype that was reversed by NMDA treatment. These data support the concept that in cerebellar slice cultures, Purkinje cell dendritic outgrowth is regulated by granule cell inputs. PMID- 15804406 TI - Phylogenetic analyses reveal deeply divergent species lineages in the genus Sphaerobolus (Phallales: Basidiomycota). AB - Phylogenetic analyses of 27 artillery fungus (Sphaerobolus sp.) isolates were conducted to identify species boundaries in the genus Sphaerobolus. Multiple gene genealogies inferred from maximum likelihood, Bayesian, and maximum-parsimony analyses of sequence data from individual loci (mtSSU, ITS, EF 1-alpha, and LSU) and a combined dataset (mtSSU, ITS, and EF 1-alpha) concordantly indicate the existence of three deeply divergent lineages in the genus Sphaerobolus, each representing a phylogenetic species. These three phylogenetic species correspond to two known species: Sphaerobolus iowensis and Sphaerobolus stellatus, and a newly discovered species. Suprageneric phylogenetic analyses of the mtSSU and LSU datasets containing representatives of related genera of the gomphoid-phalloid clade of Homobasidiomycetes suggested that the undescribed taxon likely is more closely related to S. stellatus than to S. iowensis. PMID- 15804407 TI - Molecular and morphological evolution of the amphipod radiation of Lake Baikal. AB - Lake Baikal, in Siberia, Russia, contains the highest biodiversity of any extant lake, including an impressive radiation of gammaroidean amphipods that are often cited as a classic case of adaptive radiation. However, relationships among Baikal's amphipods remain poorly understood. The phylogenetic history of 32 Lake Baikal amphipod species, representing most major lineages of the endemic fauna, was examined using three genes (COI, 16S rRNA, and 18S rRNA), and 152 morphological characters. Results support monophyly of the largest and most diverse of the Baikalian families, the Acanthogammaridae. Analyses suggest that a second Baikalian family, the fossorial Micruropodidae, is paraphyletic and composed of two divergent clades, one of which includes Macrohectopus branickii, a morphologically specialized pelagic planktivore traditionally assigned its own family. The extreme morphological and ecological divergence of Macrohectopus from its close genetic relatives, and conversely, the large genetic distances among other morphologically similar micruropodids, suggest that morphological and molecular evolution have often been uncoupled during the radiation of Baikal's amphipods. This study suggests that the amphipod fauna of Lake Baikal is polyphyletic; originating from two independent invasions of the lake. PMID- 15804408 TI - Phylogeny and biogeography of Chinese sisorid catfishes re-examined using mitochondrial cytochrome b and 16S rRNA gene sequences. AB - The family Sisoridae is one of the largest and most diverse Asiatic catfish families, most species occurring in the water systems of the Qinhai-Tibetan Plateau and East Himalayas. To date published morphological and molecular phylogenetics hypotheses of sisorid catfishes are part congruent, and there are some areas of significant disagreement with respect to intergeneric relationships. We used mitochondrial cytochrome b and 16S rRNA gene sequences to clarify existing gaps in phylogenetics and to test conflicting vicariant and dispersal biogeographical hypotheses of Chinese sisorids using dispersal vicariance analysis and weighted ancestral area analysis in combination with palaeogeographical data as well as molecular clock calibration. Our results suggest that: (1) Chinese sisorid catfishes form a monophyletic group with two distinct clades, one represented by (Gagata (Bagarius, Glyptothorax)) and the other by (glyptosternoids, Pseudecheneis); (2) the glyptosternoid is a monophyletic group and Glyptosternum, Glaridoglanis, and Exostoma are three basal species having a primitive position among it; (3) a hypothesis referring to Pseudecheneis as the sister group of the glyptosternoids, based on morphological evidence, is supported; (4) the genus Pareuchiloglanis, as presently defined, is not monophyletic; (5) congruent with previous hypotheses, the uplift of Qinghai Tibetan Plateau played a primary role in the speciation and radiation of the Chinese sisorids; and (6) an evolutionary scenario combining aspects of both vicariance and dispersal theory is necessary to explain the distribution pattern of the glyptosternoids. In addition, using a cytochrome b substitution rate of 0.91% per million years and 0.23% for 16S rRNA, we tentatively date that the glyptosternoids most possibly originated in Oligocene-Miocene boundary (19 24Myr), and radiated from Miocene to Pleistocene, along with a center of origin in the Irrawaddy-Tsangpo drainages and several rapid speciation in a relatively short time. PMID- 15804409 TI - New DNA data from a transthyretin nuclear intron suggest an Oligocene to Miocene diversification of living South America opossums (Marsupialia: Didelphidae). AB - Phylogenetic relationships of 19 species of didelphid marsupials were studied using two nuclear markers, the non-coding transthyretin intron 1 (TTR) and the coding interphotoreceptor retinoid binding protein exon 1 (IRBP), and two mitochondrial genes, the protein-coding cytochrome b (cyt-b) and the structural 12S ribosomal DNA (12S rDNA). Evolutionary dynamics of these four markers were compared to each other, revealing the appropriate properties presented by TTR intron 1 together with its well supported and resolved phylogenetic signal. Nuclear markers supported the monophyly of medium and large-sized opossums Metachirus+(Chironectes, Lutreolina, Didelphis, Philander), and the paraphyly of mouse-sized opossums, with the genera Gracilinanus, Thylamys, and Marmosops as a sister group to medium and large-sized didelphids. Conflicting branching patterns between mitochondrial and nuclear data involved the phylogenetic position of Marmosa-Micoureus-Monodelphis relative to other mouse-sized opossums. Nuclear phylogenetic inferences among genera were confirmed by the presence of synapomorphic indels observed in TTR intron 1. A Bayesian relaxed molecular clock dating of didelphid evolution using nuclear markers estimated their origin in the Middle Eocene (39.8 million years ago), with subsequent diversification during the Oligocene (Deseadan) and Miocene. PMID- 15804410 TI - Gelechioidea (Insecta: Lepidoptera) systematics: a reexamination using combined morphology and mitochondrial DNA data. AB - Phylogenies of Gelechioidea (Insecta: Lepidoptera) historically have been in disagreement and definitions vary at the family and subfamily levels. Addition of new taxa or new characters drastically changes relationships indicating that current phylogenetic schemes require more investigation. This study is the first phylogenetic analysis of Gelechioidea to include molecular data. Here we present a combined analysis using Maximum Parsimony to investigate sister-group relationships within Gelechioidea. The addition of Cytochrome oxidase I and II to revised published morphological matrices gives 453 parsimony informative characters for the 42 taxa for which we have sequence data. The combined analysis resulted in two trees with mostly novel sister-group relationships. These results challenge current concepts of Gelechioidea, suggesting that traditional morphological characters that have united taxa may not be homologous structures and are in need of further investigation. A combination of morphological data with new molecular data will be the most robust method of study for Gelechioidea phylogenetics. PMID- 15804411 TI - Phylogeny of Eleusine (Poaceae: Chloridoideae) based on nuclear ITS and plastid trnT-trnF sequences. AB - Phylogenetic relationships in the genus Eleusine (Poaceae: Chloridoideae) were investigated using nuclear ITS and plastid trnT-trnF sequences. Separate and combined data sets were analyzed using parsimony, distance, and likelihood based methods, including Bayesian. Data congruence was examined using character and topological measures. Significant data heterogeneity was detected, but there was little conflict in the topological substructure measures for triplets and quartets, and resolution and clade support increased in the combined analysis. Data incongruence may be a result of noise and insufficient information in the slower evolving trnT-trnF. Monophyly of Eleusine is strongly supported in all analyses, but basal relationships in the genus remain uncertain. There is good support for a CAIK clade (E. coracana subsp. coracana and africana, E. indica, and E. kigeziensis), with E. tristachya as its sister group. Two putative ITS homeologues (A and B loci) were identified in the allotetraploid E. coracana; the 'B' locus sequence type was not found in the remaining species. Eleusine coracana and its putative 'A' genome donor, the diploid E. indica, are confirmed close allies, but sequence data contradicts the hypothesis that E. floccifolia is its second genome donor. The 'B' genome donor remains unidentified and may be extinct. PMID- 15804412 TI - Taxonomic utility of a phylogenetic analysis of phosphoglycerate kinase proteins of Archaea, Bacteria, and Eukaryota: insights by Bayesian analyses. AB - We studied 131 protein sequences of the essentially ubiquitous glycolytic enzyme 3-phosphoglycerate kinase (3-PGK) by Bayesian analyses in three Domains: 15 Archaea, 83 Bacteria, and 33 Eukaryota. The posterior distribution of phylogenetic trees developed were based on a uniform prior, the WAG model of protein evolution, Metropolis-Hastings sampling in a Markov chain Monte Carlo analysis, and a package of diagnostics to critically evaluate the validity of the analyses. The 15 Archaea separated with high posterior probability. The archaean Phyla Euryarchaeota and the apparently Euryarchaeota derived Crenarchaeota were monophyletic. The 33 Eukaryota separated into two main groups: the non chlorophyllous forms with coherent sub-groupings of Euglenozoa, Alveolata, Fungi, and Metazoa and all the chlorophyllous species studied: the Plantae (Viridaeplantae), chlorophyllous Stramenopiles, and the chlorophyllous Bacteria. This association supports other opinions concerning the related lineage of cyanobacteria and the Plantae. The 3-PGK sequences from 83 Bacteria in almost every instance associated by their recognized taxal group: alpha-, beta-, gamma-, epsilon-proteobacteria, Chlamydia, Actinobacteridae, and Firmicutes. Firmicutes sequences were subdivided into three apparently monophyletic groups: the anaerobic Clostridia, the spore-forming Bacillales and a group containing the Mollicutes, Lactobacillales and non-spore-forming Bacillales. The 3-PGK-gene tree assemblage was notable both for its pervasive clustering in three Domains according to recognized taxonomic groupings of Class, Order, Family, and Genus. The 3-PGK enzyme or 3-PGK-like activity may have played a central role in the metabolism of the Universal Ancestor. PMID- 15804413 TI - Phylogeny of Tubificidae (Annelida, Clitellata) based on mitochondrial and nuclear sequence data. AB - The tubificid clitellates are a common component in the freshwater bottom fauna and are also the most abundant oligochaete group in marine habitats. There are over 800 described species classified in six subfamilies; Tubificinae, Limnodriloidinae, Rhyacodrilinae, Telmatodrilinae, Phallodrilinae, and Naidinae. In this study we examine the phylogenetic relationships in Tubificidae using a combination of mitochondrial 16S rDNA and nuclear 18S rDNA sequence data. Sequences were obtained from five outgroup and 56 ingroup taxa, including five of the six subfamilies of Tubificidae. The data were analysed by maximum parsimony and Bayesian inference. The resulting tree topologies are virtually without conflict. Several associations traditionally recognized within the family Tubificidae are supported, in the Bayesian analysis including a sister group relationship between Tubificinae and Limnodriloidinae. The results also indicate that Rhyacodrilinae is polyphyletic--some of its members (Heterodrilus spp.) fall into a clade with Phallodrilinae, all other groups with Naidinae. Naidinae is also polyphyletic with two rhyacodriline genera, Monopylephorus and Ainudrilus, nested within. Most of the tubificid genera included in the study are supported as monophyletic; however, Tubifex and Limnodriloides are refuted, and Tubificoides is unresolved from other tubificine taxa. PMID- 15804414 TI - A molecular phylogeny of the hawkmoth genus Hyles (Lepidoptera: Sphingidae, Macroglossinae). AB - The hawkmoth genus Hyles is one of 15 genera in the subtribe Choerocampina of the subfamily Macroglossinae. Due to a remarkable uniformity, morphological characters usually used to identify and classify Lepidoptera at the species level cannot be used in this genus. Instead, we used DNA sequences comprising about 2300 bp derived from the mitochondrial genes COX I, COX II, and tRNA-leucine to elucidate the phylogeny of Hyles. The results corroborate the monophyly of Hyles but conflict with previous internal classifications of the genus based on morphology. Hyles seems to have evolved in the Neotropics during the Oligocene/Eocene epochs and the molecular data (which evolved clock-like) confirm the hypothesis that it is a very young genus that radiated on a global scale rather quickly. We hypothesize its sister group to be one of the genera Deilephila, Theretra or Xylophanes. The Nearctic may have been colonized rapidly by Hyles once the land bridge formed during the Pliocene, since within this same Epoch, the invasion of the Palaearctic appears to have proceeded from the East, via the Bering route. The colonization of Australia appears to have occurred rather early in Hyles radiation, although the route is not clear. We propose that the radiation of the Hyles euphorbiae-complex s. str. (HEC) occurred as recently as the Pliocene/Pleistocene boundary and that its roots can still be reconstructed in Asia. Hyles dahlii is closely related to the HEC, but a sister group relationship to the HEC s. str. cannot be corroborated unequivocally. HEC population ranges appear to have tracked climate oscillations during the Pleistocene Ice Ages, resulting in hybridization around the Mediterranean Sea as they repeatedly intermingled. Comparison of the phylogeny with food plant affiliations leads us to hypothesize that Euphorbia monophagy evolved at least two times independently within Hyles. PMID- 15804415 TI - A phylogenetic framework for the terns (Sternini) inferred from mtDNA sequences: implications for taxonomy and plumage evolution. AB - We sequenced 2800 bp of mitochondrial DNA from each of 33 species and 2 subspecies (35 taxa) of terns (Sternini), and employed Bayesian methods to derive a phylogeny with good branch support based on posterior probabilities. The resulting tree confirmed many of the generally accepted taxonomic groups, and led us to suggest a revision of the terns that recognizes 12 genera, 11 of which correspond to a distinct clade on the tree or a highly divergent species (1 genus was not represented in the phylogeny). As an example of how the molecular phylogeny reflects similarities in morphology and behavior among the terns, we used the phylogeny to examine the evolution of the breeding (alternate) head plumage patterns among the terns to test the hypothesis that this character is phylogenetically informative. The three basic types of head plumage (white crown, black cap, and black cap with a white blaze on the forehead) were highly conserved within clades, with notable exceptions in two white-crowned species that evolved independently among the black-capped terns. Based on the appearance of the close relatives of these exceptional species, their white crowns appear to be due to the retention of either winter (basic) plumage characteristics or perhaps juvenile characteristics when the birds molt into their breeding plumage. Examination of the evolutionary history of head plumage indicated that the white crowned species such as the noddies (Anous) and the white tern (Gygis alba) are probably most representative of ancestral terns. PMID- 15804416 TI - The repertoire of trace amine G-protein-coupled receptors: large expansion in zebrafish. AB - Trace amines, such as tyramine, beta-phenylethylamine, tryptamine, and octopamine, are present in trace levels in nervous systems and bind a specific family of G-protein-coupled receptors (GPCR), but the function or origin of this system is not well understood. We searched the genomes of several eukaryotic species for receptors similar to the mammalian trace amine (TA) receptor subfamily. We identified 18 new receptors in rodents that are orthologous to the previously known TA-receptors. Remarkably, we found 57 receptors (and 40 pseudogenes) of this type in the zebrafish (Danio rerio), while fugu (Takifugu rubripes) had only eight receptors (and seven pseudogenes). We mapped 47 of the zebrafish TA-receptors on chromosomes using radiation hybrid panels and meiotic mapping. The results, together with the degree of conservation and phylogenetic relationships displayed among the zebrafish receptors suggest that the family arose through several different mechanisms involving tetraploidization, block duplications, and local duplication events. Interestingly, these vertebrate TA receptors do not show a close evolutionary relationship to the invertebrate TA binding receptors in fruitfly (Drosophila melanogaster), indicating that the ability to bind TA have evolved at least twice in animal evolution. We collected in total over 100 vertebrate TA-receptor sequences, and our phylogenetic analysis shows that several TA-receptors have evolved rapidly with remarkable species variation and that the common ancestor of vertebrate TA-receptors arose before the split of the ray-finned and lobe-finned fishes. The evolutionary history of the TA-receptors is more complex than for most other GPCR families and here we suggest a mechanism by which they may have arisen. PMID- 15804417 TI - Phylogenetic studies of pantherine cats (Felidae) based on multiple genes, with novel application of nuclear beta-fibrinogen intron 7 to carnivores. AB - The pantherine lineage of the cat family Felidae (order: Carnivora) includes five big cats of genus Panthera and a great many midsized cats known worldwide. Presumably because of their recent and rapid radiation, the evolutionary relationship among pantherines remains ambiguous. We provide an independent assessment of the evolutionary history of pantherine lineage using two complete mitochondrial (mt) genes (ND2 and ND4) and the nuclear beta-fibrinogen intron 7 gene, whose utility in carnivoran phylogeny was first explored. The available four mt (ND5, cytb, 12S, and 16SrRNA) and two nuclear (IRBP and TTR) sequence loci were also combined to reconstruct phylogeny of 14 closely related cat species. Our analyses of combined mt data (six genes; approximately 3750 bp) and combined mt and nuclear data (nine genes; approximately 6500 bp) obtained identical tree topologies, which were well-resolved and strongly supported for almost all nodes. Monophyly of Panthera genus in pantherine lineage was confirmed and interspecific affinities within this genus revealed a novel branching pattern, with P. tigris diverging first in Panthera genus, followed by P. onca, P. leo, and last two sister species P. pardus and P. uncia. In addition, close association of Neofelis nebulosa to Panthera, the phylogenetic redefinition of Otocolobus manul within the domestic cat group, and the relatedness of Acinonyx jubatus and Puma concolor were all important findings in the resulting phylogenies. The potential utilities of nine different genes for phylogenetic resolution of closely related pantherine species were also evaluated, with special interest in that of the novel nuclear beta-fibrinogen intron 7. PMID- 15804418 TI - Origin, diversification, and historical biogeography of the genus Trachurus (Perciformes: Carangidae). AB - We addressed phylogenetic relationships in the genus Trachurus using cytochrome b gene and D-loop sequences. The trees showed five groups: (1) the Southwest Pacific species (T. japonicus, T. novaezelandiae, and T. declivis); (2) The Mediterranean Sea and Eastern Atlantic species (T. mediterraneus); (3) The Atlantic Ocean species (T. lathami and T. trecae); (4) Eastern Atlantic species (T. trachurus and T. capensis); and (5) a group of highly mobile pelagic species, two from the Eastern Pacific (T. symmetricus and T. murphyi) and one from the Eastern Atlantic (T. picturatus). The phylogeny based on Cyt b, supports the molecular clock hypothesis and our results agree with the reported fossil indicating that the origin of this genus occur when the Thetys Sea closed (around 18.4 MYA). In addition, a very slow neutral substitution rate is reported identified only two periods of maximum diversification: the first occurring between 18.4 and 15.0 MYA and the second between 8.4 MYA and present day. PMID- 15804419 TI - Protein diversity is generated within the motin family of proteins by alternative pre-mRNA splicing. AB - The motin family of proteins is comprised of three polypeptides, angiomotin, angiomotin-like 1, and angiomotin-like 2. Angiomotin is an angiostatin-binding protein that promotes endothelial cell motility and is involved in angiogenesis. The function of human angiomotin-like-1 and angiomotin-like-2, however, remains unknown. In this report, we investigated the potential for molecular diversity within the motin family of proteins through the identification and characterization of alternatively spliced transcripts in endothelial cells, human tissues and a variety of cell lines. We report that the motins display variability at the mRNA level suggesting an intricate regulatory system at the transcriptional and potentially protein level. Some alternative transcripts are expressed in a tissue-specific manner and others give rise to novel protein isoforms. The alternative splicing that occurs within this protein family may have important implications in the regulation of the expression and function of the motins. PMID- 15804420 TI - Neurofibromin and its inactivation of Ras are prerequisites for osteoblast functioning. AB - Skeletal problems and osteoporosis occur in up to 50% affected neurofibromatosis type 1 (NF1) humans. Inactivation of neurofibromin results in deregulation of Ras signal transduction. Little is known of bone biology in humans with NF1. The goal of our work was to determine if loss-of-function of Nf1 gene was associated with altered bone homeostasis and Ras signal transduction. Because homozygous Nf1 mice are embryonically lethal, heterozygote Nf1 (Nf1+/-) male mice were used to investigate skeletal phenotypes and osteoprogenitor functions, using standard in vivo and in vitro assays. We found that bone mass and geometry of Nf1+/- mice did not differ from wild type controls, despite a trend to less bone formation. Nf1+/ committed osteoprogenitors from femur metaphysis exhibited premature apoptosis and higher proliferation. Ras signaling was activated in primary Nf1+/- bone marrow-inducible osteoprogenitors. Inducible osteoprogenitors exhibited lower induction of osteoblast differentiation, assessed as alkaline phosphatase positive CFU-f. A screen of osteoblast marker genes showed a selective increase in osteopontin (OPN) mRNA and protein expression in these cells. OPN protein was increased in Nf1+/- bone, especially in cortical bone matrix. Because bone cell abnormalities in Nf1 haploinsufficiency were detected in vitro, redundant pathways must compensate for the deregulation of Ras signaling in vivo to maintain normal bone mass and function in vivo. Our in vitro data revealed that neurofibromin and its control of Ras signaling are required for osteoprogenitor homeostasis. PMID- 15804421 TI - Hyperpolarization-activated current (Ih): a characterization of subicular neurons in brain slices from socially and individually housed rats. AB - The hyperpolarization and cyclic nucleotide activated current Ih is thought to have a role in rhythmic brain activity that is important in complex behaviors and might be perturbed in some neuropsychiatric diseases. We have used whole-cell voltage and current clamp techniques to characterize Ih in neurons from the subiculum-the major output region of the hippocampal formation. Subicular projection neurons are themselves classifiable as intrinsically bursting (IB) or regular spiking (RS) and Ih is present in both. Given the possible involvement of Ih in neuropsychiatric diseases, we have also characterized Ih in subicular neurons from rats that have been housed in individual cages (though still able to see, smell, and hear other rats) as these rats can display behavioral changes similar to those seen in schizophrenia. Individual housing is associated with a 4.4-mV depolarization of the Ih activation curve (P=0.0027) and an increase in mean firing rate measured in response to current injection (P=0.037) specifically in RS neurons and a change in the relative amplitude of Ih between IB and RS neurons. Thus, we have shown significant changes in a current thought to be relevant to psychiatric disease in a partial model of schizophrenia. Its further investigation might reveal chemical targets for novel antipsychotic drugs. PMID- 15804422 TI - Evidence that urocortin I acts as a neurohormone to stimulate alpha MSH release in the toad Xenopus laevis. AB - We have raised the hypothesis that in the South African clawed toad Xenopus laevis, urocortin 1 (UCN1), a member of the corticotropin-releasing factor (CRF) peptide family, functions not only within the brain as a neurotransmitter/neuromodulator but also as a neurohormone, promoting the release of alpha-melanophore-stimulating hormone (alphaMSH) from the neuroendocrine melanotrope cells in the intermediate lobe of the pituitary gland. This hypothesis has been investigated by (1) assessing the distribution of UCN1 and CRF by light immunocytochemistry, (2) determining the subcellular presence of UCN1 in the neural lobe of the pituitary gland by immuno-electron microscopy applying high-pressure freezing and cryosubstitution, and (3) testing the effect of UCN1 on MSH release from toad melanotrope cells using in vitro superfusion. In the X. laevis brain, the main site of UCN1-positive somata was found to be the Edinger-Westphal nucleus. UCN1 immunoreactivity (ir) also occurs in the nucleus posteroventralis tegmenti, central gray, nucleus reticularis medius, nucleus motorius nervi facialis, and nucleus motorius nervi vagi. UCN1 occurs together with CRF in the nucleus motorius nervi trigemini, and in the magnocellular nucleus, which send a UCN1- and CRF-containing fiber tract to the median eminence. Strong UCN1-ir and CRF-ir were found in the external zone of the median eminence. From the internal zone of the median eminence, UCN1-ir fibers, but few CRF-ir fibers, were found to project to the pituitary neural lobe, where they form numerous neurohemal axon terminals. Ultrastructurally, two types of terminal containing UCN1-ir secretory granules were distinguished: type A contains large, moderately electron-dense, round secretory granules and type B is filled with smaller, strongly electron-dense, ellipsoid secretory granules. In vitro superfusion studies showed that UCN1 stimulated the release of alphaMSH from melanotrope cells in a dose-dependent manner. Our results support the hypothesis that in X. laevis, UCN1 released from neurohemal axon terminals in the pituitary neural lobe functions as a stimulatory neurohormone for alphaMSH release from melanotrope cells of the pituitary intermediate lobe. PMID- 15804423 TI - Intracellular oxidation of allopregnanolone by human brain type 10 17beta hydroxysteroid dehydrogenase. AB - Allopregnanolone is a positive allosteric modulator of GABAA receptors, generated by the reduction of 5alpha-dihydroprogesterone (5alpha-DHP) in astrocytes. This neuroactive steroid can be inactivated by its 3alpha-oxidation to yield 5alpha DHP. It was found that 5alpha-DHP levels in HEK293 cells expressing type 10 17beta-hydroxysteroid dehydrogenase (17beta-HSD10), but not its catalytic inactive mutant, increased significantly as allopregnanolone was added to culture media. The results demonstrate that mitochondrial 17beta-HSD10 effectively catalyzes the intracellular oxidation of allopregnanolone. Moreover, brain astrocytes contain a moderate level of 17beta-HSD10, which is elevated in activated astrocytes of brains with Alzheimer type pathology, including sporadic Alzheimer's disease (AD) and Down's syndrome with AD. The distribution of 17beta HSD10 was found not to parallel that of 3alpha-HSD3. Cerebral cortex has the lowest level of 17beta-HSD10; whereas the hippocampus, hypothalamus, and amygdala possess relatively higher levels of this enzyme. The catalysis of 17beta-HSD10 appears to be essential for maintaining normal functions of GABAergic neurons. The elevated level of 17beta-HSD10 in activated astrocytes is a new feature found in brains of people with AD, and it may have important impact on AD pathogenesis. PMID- 15804424 TI - Spatial resolution of calpain-catalyzed proteolysis in focal cerebral ischemia. AB - Transient forebrain ischemia induces calpain-mediated degradation of the neuronal cytoskeleton, alpha-fodrin, and this results in ischemic neuronal death. In this study, we investigated the spatial distribution and temporal changes of calpain catalyzed alpha-fodrin proteolysis in focal cerebral ischemia and examined the effects of a calpain inhibitor. Ischemia was induced in gerbils by 3-h middle cerebral artery occlusion followed by reperfusion. Animals were divided into four groups: a sham-operated group, an ischemic group, a vehicle-treated group, and a calpain inhibitor-treated group. Intravenous injections of vehicle or calpain inhibitor I were administered 30 min before ischemia. Infarct volumes were measured 1 day after reperfusion and the spatial distribution of calpain catalyzed alpha-fodrin proteolysis was investigated by immunohistochemistry 15 min, 1 h, 4 h, and 1 day after reperfusion. Infarct volume (mean +/- SD) in the ischemic group and the vehicle-treated group was 204.6 +/- 19.1 mm3 and 212.4 +/- 16.3 mm3, respectively, and the calpain inhibitor I reduced the infarct volume [149.4 +/- 25.2 mm3 (P < 0.05)]. Immunoblot analysis demonstrated that calpain inhibitor reduced proteolysis. Ischemia induced fodrin proteolysis in the ischemic core and the peri-infarct zone within 15 min after reperfusion, with proteolysis developing quickly in the ischemic core and more slowly in the peri infarct zone. Proteolysis preceded neuronal death in the peri-infarct zone. Calpain inhibitor I ameliorated neuronal death in the peri-infarct zone but not in the ischemic core. Thus, calpain plays a pivotal role on focal ischemia as well as in global ischemia. PMID- 15804425 TI - Fatigue induced by intermittent maximal voluntary contractions is associated with significant losses in muscle output but limited reductions in functional MRI measured brain activation level. AB - The main purpose of this study was to characterize brain activation patterns during a fatigue task involving repetitive maximal voluntary contractions (MVC) of finger flexor muscles. Fourteen young, healthy human participants performed approximately 100 handgrip MVCs (each 2-s contraction was followed by a 1-s rest) while their brain was imaged by functional MRI (fMRI). The handgrip force and electromyograms (EMG) of the finger flexors declined progressively to about 40% of the initial values at the end of the fatigue task, suggesting that significant muscle fatigue had occurred. In contrast, the level of the fMRI signal in the primary (sensorimotor), secondary (supplementary motor), and association (prefrontal and cingulate) motor-function cortices did not change significantly throughout the fatigue task (although the signal of the primary sensorimotor cortex showed a clear trend of decline). The fMRI data from the task of intermittent handgrip MVCs differed dramatically from those obtained in a 2-min sustained handgrip MVC published in a recent report, in which the overall fMRI measured brain activation level was substantially lower and followed an increase then-decrease pattern compared to the linear decreases in force and EMG. These results support the notion that the motor cortical centers control the tasks of repetitive and continuous muscle contractions differently and that there is a decoupling in the signal changes of the brain and muscles during muscle fatigue processes induced by maximal voluntary contractions. PMID- 15804426 TI - Effect of chronic intermittent restraint stress on hippocampal expression of marker proteins for synaptic plasticity and progenitor cell proliferation in rats. AB - Chronic restraint stress may change hippocampal mRNA levels of markers for synaptic plasticity such as synaptophysin, growth-associated protein 43 (GAP-43), and brain-derived neurotrophic factor (BDNF). In order to examine the relation between that stressor and those biochemical markers on protein level as well as the Ki-67 protein, a marker of progenitor cell proliferation, we subjected rats to chronic intermittent restraint stress for 6 h per day for 14 days excluding the weekends. This stress intensity caused a significant increase in adrenal gland weight and decrease in body weight gain. However, we did not find significant alteration of protein expression levels for synaptophysin, GAP-43, and BDNF by using Western blot analysis. Unlike these findings, the hippocampal protein expression of Ki-67 was significantly reduced by using both Western blot and immunohistochemical analyses. This reduction of Ki-67 expression in chronically stressed rats was correlated with increased adrenal gland weight and decreased body weight gain. All marker proteins used did not show any changes of hippocampal expression level after a single restraint stress session of 3 h. In conclusion, chronic intermittent restraint stress caused changes in the physiological stress response in rats, and a decrease of hippocampal progenitor cells using the Ki-67 protein as marker which indicates a suppression of adult neurogenesis. The results might contribute to understand the relationship between stress and cellular neurobiology of depression, since chronic antidepressant treatment have been shown to increase adult neurogenesis in the rat hippocampus. PMID- 15804427 TI - Effect of the activation of central 5-HT2C receptors by the 5-HT2C agonist mCPP on blood pressure and heart rate in rats. AB - In the present study we investigated the role of central 5-HT2C receptors in the control of blood pressure and heart rate in non-stressed and stressed, adult, male, Wistar rats. Third ventricle injections of the 5-HT2C agonist mCPP elicited a significant increase in blood pressure in non-stressed animals. The initial period of this hypertensive response (10-30 min after mCPP administration) was accompanied by baroreflex-mediated bradycardia, while after this period the coexistence of hypertension and tachycardia was observed. These cardiovascular effects promoted by the central administration of mCPP were blocked by pretreatment with the 5-HT2C antagonist, SDZ SER 082. The administration of SDZ SER 082 alone induced no significant changes in blood pressure or heart rate. The pharmacological stimulation of central 5-HT2C receptors by mCPP did not change the hypertensive or tachycardic responses induced by restraint stress. Conversely, the blockade of central 5-HT2C receptors by SDZ SER 082 blunted stress-induced hypertension without modifying stress-induced tachycardia. It is concluded that the activation of central 5-HT2C receptors induces hypertension in non-stressed rats and that the normal function of these receptors is essential for the rise in blood pressure that occurs in the course of restraint stress. PMID- 15804428 TI - Alpha-synuclein-positive structures induced in leupeptin-infused rats. AB - Abnormal accumulation of alpha-synuclein is regarded as a key pathological step in a wide range of neurodegenerative processes, not only in Parkinson's disease (PD) and dementia with Lewy bodies (DLB) but also in multiple-system atrophy (MSA). Nevertheless, the mechanism of alpha-synuclein accumulation remains unclear. Leupeptin, a protease inhibitor, has been known to cause various neuropathological changes in vivo resembling those of aging or neurodegenerative processes in the human brain, including the accumulation of neuronal processes and neuronal cytoskeletal abnormalities leading to neurofibrillary tangle (NFT) like formations. In the present study, we administered leupeptin into the rat ventricle and found that alpha-synuclein-positive structures appeared widely in the neuronal tissue, mainly in neuronal processes of the fimbria and alveus. Immunoelectron microscopic study revealed that alpha-synuclein immunoreactivity was located in the swollen axons of the fimbria and alveus, especially in the dilated presynaptic terminals. In addition colocalization of alpha-synuclein with ubiquitin was rarely observed in confocal laser-scan image. This is the first report of experimentally induced in vivo accumulation of alpha-synuclein in non transgenic rodent brain injected with a well-characterized protease inhibitor by an infusion pump. The present finding suggests that the local accumulation of alpha-synuclein might be induced by the impaired metabolism of alpha-synuclein, which are likely related to lysosomal or ubiquitin-independent proteasomal systems. PMID- 15804429 TI - GFAP-expressing radial glia-like cell bodies are involved in a one-to-one relationship with doublecortin-immunolabeled newborn neurons in the adult dentate gyrus. AB - The present study examined the relationship between radial glial cells and newborn neurons in the adult dentate gyrus using three different methods. Single labeling immunocytochemistry for newly born neurons using doublecortin, as well as double labeling using an additional antibody to glial fibrillary acidic protein (GFAP) to label astrocytes were used at the light microscopic level. Furthermore, doublecortin immunoelectron microscopy was used to examine the ultrastructural relationship between newborn neurons and astrocytes in the adult dentate gyrus. These data showed an intimate one-to-one relationship between GFAP expressing radial glia-like cell bodies and their non-radial processes that wrap around the basal and lateral sides of newborn neurons to cradle them in the subgranular zone. A similar relationship is observed for the newborn neurons at the base of the granule cell layer, but the cell body of the GFAP-expressing radial glia-like cells is not as intimately associated with the cell body of the newborn neurons at this site. Furthermore, newborn neurons with apical dendritic processes and growth cones in the granule cell layer extend them along radial glial processes. These newborn neurons do not receive axosomatic or axodendritic synapses indicating the absence of basket cell innervation. These data show that GFAP-expressing radial glia-like cells in the dentate gyrus cradle newborn neurons in the subgranular zone and that their radial processes provide a scaffold for neuronal process outgrowth. PMID- 15804430 TI - Tetramethylpyrazine inhibits ATP-activated currents in rat dorsal root ganglion neurons. AB - Tetramethylpyrazine (TMP) is one of the alkaloids contained in Ligustrazine which has been used in traditional Chinese medicine as an analgesic for injury and dysmenorrhea. ATP can elicit the sensation of pain. This study observed the effects of TMP on ATP-activated current (IATP) in rat DRG neurons. TMP (0.1-1 mM) concentration-dependently inhibited ATP (100 microM)-activated current in rat DRG neurons. The inhibitory time of ATP (100 microM)-activated current appeared at 15 s after preapplication of TMP and reached its peak at about 45 s. The dose response curves for IATP in the absence and presence of 1 mM TMP showed that TMP (1 mM) shifted the concentration-response curve of IATP downward markedly and the two EC50 values were very close (75 vs. 82 microM), while the threshold value remained unchanged. Therefore, the inhibitory effect of TMP on IATP may be noncompetitive. TMP did not alter the reversal potential (0 mV) of ATP-activated current, indicating that the site of TMP action is on or near the exterior surface of channel protein and not within the channel pore. Externally applied TMP (1 mM) increases the inhibitory effect of chelerythrine (PKC inhibitor) contained in pipette solution on IATP. The site of TMP action may be the binding of TMP to an allosteric site on the large extracellular region of ATP receptor ion channel complex (P2X receptors) or PKC site of the N-terminus of P2X receptors. The mechanism of TMP action may be the allosteric regulation via acting on the large extracellular region of ATP receptor-ion channel complex (P2X receptors) and promoting the phosphorylation of PKC site of the N-terminus of P2X receptors. PMID- 15804431 TI - Adult retinal pigment epithelium cells express neural progenitor properties and the neuronal precursor protein doublecortin. AB - The adult mammalian retina is devoid of any detectable neurogenesis. However, different cell types have been suggested to potentially act as neural progenitors in the adult mammalian retina in vitro, such as ciliary body (CB), Muller glia, and retinal pigment epithelium (RPE) cells. In rodents and humans, strong evidence for neural stem or progenitor properties exists only for CB-derived cells, but not for other retinal cell types. Here, we provide a comparative analysis of adult rat CB- and RPE-derived cells suggesting that the two cell types share certain neural progenitor properties in vitro. CB and RPE cells expressed neural progenitor markers such as Nestin, Flk-1, Hes1, and Musashi. They proliferated under adherent and neurosphere conditions and showed limited self-renewal. Moreover, they differentiated into neuronal and glial cells based on the expression of differentiation markers such as the young neuronal marker beta-III tubulin and the glial and progenitor markers GFAP and NG2. Expression of beta-III tubulin was found in cells with neuronal and non-neuronal morphology. A subpopulation of RPE- and CB-derived progenitor cells expressed the neurogenesis specific protein doublecortin (DCX). Interestingly, DCX expression defined a beta III tubulin-positive CB and RPE fraction with a distinct neuronal morphology. In summary, the data suggest that RPE cells share with CB cells the potential to de differentiate into a cell type with neural progenitor-like identity. In addition, DCX expression might define the neuronal-differentiating RPE- and CB-derived progenitor population. PMID- 15804432 TI - Susceptibility to excitotoxic and metabolic striatal neurodegeneration in the mouse is genotype dependent. AB - Previously, we had reported that hippocampal susceptibility to the neurotoxic effects of excitotoxin administration is strain dependent [Schauwecker and Steward, Proc. Natl. Acad. Sci. U.S.A. 94 (1997) 4103]. However, it has been unclear whether strain-related gene products may play a similar role in providing protection against drugs that produce striatal lesions. The present series of experiments sought to elucidate whether genetic background alters neuronal viability within the striatum following metabolic or excitotoxic injury. Thus, we have examined the effect of mouse strain on susceptibility to striatal injury using well-characterized animal models of Huntington's disease by examining whether C57BL/6 mice, previously identified as resistant to excitotoxin-induced hippocampal cell death, are resistant to quinolinate, malonate, and 3 nitropropionic acid (3-NP). Intrastriatal injection of either malonate or quinolinate and systemic administration of 3-NP resulted in significantly smaller striatal lesions in C57BL/6 mice as compared to FVB/N mice, previously identified as susceptible to hippocampal excitotoxic injury. The existence of an animal strain with decreased resistance to striatal lesions suggests that there are mediating factors involved in the preferential vulnerability of the striatum to neurotoxic lesioning. The identification of these factors could provide strategies for therapeutic intervention in Huntington's disease. PMID- 15804433 TI - Cardiovascular effects of angiotensin II and angiotensin-(1-7) at the RVLM of trained normotensive rats. AB - We evaluated the effect of physical training on the cardiovascular responses produced by angiotensin peptides at the rostroventrolateral medulla (RVLM) of non anesthetized normotensive rats. The RVLM pressor effect induced by Ang II was significantly greater in trained rats, while, in contrast, the Ang-(1-7) pressor effect was significantly smaller in trained in comparison to sedentary rats. In addition, the RVLM microinjection of Losartan (AT1 Ang II receptor antagonist) or A-779 (Ang-(1-7) receptor antagonist) induced opposite effect in trained rats. These results show that exercise training induces a differential RVLM responsiveness to Ang peptides, which was corroborated by the selective antagonists, indicating that the RVLM is a site in the central nervous system involved in the adaptive mechanisms triggered during exercise training. PMID- 15804434 TI - Role of protein kinase C beta in phorbol ester-induced c-fos gene expression in neurons of normotensive and spontaneously hypertensive rat brains. AB - We have previously demonstrated that pressure application of the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) onto some neurons in the anterior hypothalamic area of rats increases neural activity in vivo and that this PKC activation-induced increase of neural activity is enhanced in spontaneously hypertensive rats (SHR), an animal model for genetic hypertension. Activation of PKC increases expression of the c-fos gene, an important transcription factor and proto-oncogene thought to be a marker of neural activity. To evaluate PKC isoforms responsible for neural activation, we examined which isoforms of PKC are involved in the PKC activation-induced c-fos gene expression in neuronal cultures of Wistar rat and spontaneously hypertensive rat (SHR) brains. PMA increased c-fos gene expression in neuronal cultures of Wistar rat brain and the PMA-induced c-fos gene expression was inhibited by the PKC inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7). The PKCalpha,beta,gamma activator thymeleatoxin also increased c-fos gene expression, while the PKCdelta,epsilon activator ingenol did not affect it. In addition, the PMA-induced c-fos gene expression was inhibited by PKCbetaantisense oligonucleotides (AON) but not by PKCalpha and PKCgammaAONs. In SHR brain neuronal cultures, the PMA-induced c-fos gene expression was enhanced as compared with that of Wistar Kyoto rats (WKY), while basal c-fos gene expression was almost the same in both neuronal cultures. The enhancement of PMA-induced c-fos gene expression in SHR brain cultures was abolished by PKCbetaAON. These findings suggest that in rat brain neuronal cultures, PMA increases c-fos gene expression via activation of PKC and that PKCbetaisoforms are partly involved in the PMA induced c-fos gene expression. In neuronal cultures of SHR brain, it appears that the PMA-induced c-fos gene expression is also enhanced via PKCbeta. PMID- 15804436 TI - Selective potentiation of opioid analgesia by nonsteroidal anti-inflammatory drugs. AB - Opioids are often used in conjunction with nonsteroidal anti-inflammatory drugs (NSAIDs) in the treatment of moderate to severe pain. In this study we have examined interactions between these two classes of drugs. NSAIDs are inactive in the radiant heat tail-flick test, an assay of moderate to severe pain in which opioids are effective. In this assay, ibuprofen potentiated the analgesic actions of hydrocodone and oxycodone, shifting their ED(50) values by 2.5-fold and 4.6 fold despite its inactivity when given alone. These opioid/NSAID interactions were dependent upon both the opioid and the NSAID. Neither aspirin nor ketorolac influenced hydrocodone actions in this model and ibuprofen did not potentiate fentanyl or morphine analgesia. Together, these studies demonstrate potent interactions between selected combinations of opioids and NSAIDS and may help explain the clinical utility of combinations. However, the findings also illustrate differences between the drugs within each class. PMID- 15804435 TI - Newborn neurons acquire high levels of reactive oxygen species and increased mitochondrial proteins upon differentiation from progenitors. AB - A population of embryonic rat cortical cells cultured in the presence of FGF2 and having neuronal morphology expressed higher levels of reactive oxygen species (ROS) than did progenitor cells, astrocytes, and several cell lines of neuronal and non-neuronal origin. ROS were assessed using 5-(and-6)-chlormethyl-2',7' dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCF-DA), and high levels persisted in the presence of antioxidants or lowered levels of ambient oxygen. Greater than 95% of high ROS-producing cells, isolated by fluorescence-activated cell sorting, expressed the neuronal marker beta III tubulin. These cells did not incorporate BrdU or express nestin, unlike low ROS-producing cells, 99% of which exhibited both of these characteristics. Upon growth factor removal, low ROS expressing cells differentiated into neurons and astrocytes and these neurons expressed high levels of ROS, indicating that ROS accumulation accompanies the differentiation of progenitors into neurons. ROS levels were decreased by added superoxide dismutase and catalase, suggesting that both superoxide and hydrogen peroxide contribute to the ROS signal. High ROS-expressing cells also contained higher levels of several mitochondrial respiratory chain components. Although ROS have been associated with conditions that lead to cell death, our results and recent studies on the role of ROS as regulators of signal pathways are consistent with the possibility that ROS play a role in the development of the neuronal phenotype. Moreover, the differential production of ROS provides a useful method to isolate from mixed populations cells that are highly enriched for either progenitor cells or neurons. PMID- 15804437 TI - Fast neurotransmission in the rat medial preoptic nucleus. AB - The functional properties of neurotransmission in the medial preoptic nucleus (MPN) were studied in a brain slice preparation from young male rats. The aims were to evaluate the thin slice preparation for studying evoked synaptic responses in MPN neurons, to characterize the fast responses triggered by activation of presynaptic nerve fibers in the MPN, and to identify the involved receptor types. Presynaptic stimulation within the MPN evoked postsynaptic voltage and current responses that were blocked by 200 microM Cd2+ or by 2.0 microM tetrodotoxin and were attributed to action potential-evoked transmitter release. The relation to stimulus strength and comparison with spontaneous synaptic currents suggested that in many cases only one presynaptic nerve fiber was excited by the stimulus. Furthermore, the transmission was probabilistic in nature, with frequent failures. Thus, response probability, most likely reflecting transmitter release probability, could be evaluated in the thin slice preparation. Evoked excitatory postsynaptic currents recorded under voltage-clamp conditions were, due to kinetics, I-V relation, and pharmacological properties, attributed to AMPA/kainate receptors and NMDA receptors, whereas inhibitory currents were attributed to GABAA receptors. No responses that could be attributed to glycine or other types of primary transmitters were detected. Although serotonin (5-HT) did not appear to function as a primary transmitter, glutamate- as well as GABA-mediated transmission was suppressed by 500 microM 5 HT, with a clear reduction in response probability observed. 5-HT also reduced the frequency, but not the amplitude, of spontaneous postsynaptic currents and was therefore ascribed a presynaptic site of action. PMID- 15804438 TI - CREB phosphorylation following hypoxia and ischemia in striatum of newborn piglets: possible role of dopamine. AB - The goal of the present study was to determine the effects of hypoxia and ischemia and the role of dopamine on phosphorylation of cAMP response element binding protein (CREB) in striatum of newborn piglets. Piglets, with and without prior injection of alpha-methyl-p-tyrosine (AMT), an inhibitor of dopamine (DA) synthesis, were subjected to 1 h of hypoxia (decreased inspired oxygen pressure, FiO2, from 21 to 6%) or 1 h of ischemia (ligation of both carotid arteries and hemorrhage to reduce the systemic arterial pressure to about 40 mmHg), followed by 2 h recovery. Microvascular oxygen pressure in the cortex (pCO2) was measured by quenching of phosphorescence. Extracellular DA was determined by in vivo microdialysis. Striatal levels of phosphorylated CREB (pCREB) and total CREB were determined by Western blots. In sham-operated animals, pCO2 was 49.7 +/- 8.2 mmHg. During hypoxia and ischemia, pCO2 decreased to 6.3 +/- 1.8 mmHg and 10.2 +/ 2.7 mmHg, respectively. There was statistical difference in the level of extracellular DA during hypoxia versus ischemia. At the end of ischemia and hypoxia, the levels of DA were 96 x 10(3) +/- 24 x 10(3)% and 26 x 10(3) +/- 12 x 10(3)% of control, respectively. The pCREB measured after 2 h recovery was not changed after hypoxia but was decreased to 47.8 +/- 24% of control after ischemia. Depletion of endogenous DA abolished the ischemia-induced decrease in pCREB level. Total CREB did not change after either condition. It can be concluded that observed decreases of CREB phosphorylation following ischemia can be at least partially due to the high extracellular DA level. PMID- 15804439 TI - Immunohistochemical study of p47Phox and gp91Phox distributions in rat brain. AB - NADPH oxidase is multi-component enzyme, which comprises the cytosolic proteins p40Phox, p47Phox, and p67Phox and the two membrane proteins, gp91Phox and p22Phox, and which is well characterized in phagocytic cells. NADPH oxidase is a primary source of reactive oxygen species (ROS), and recent studies indicate that free radicals and ROS might be causative factors of several brain degenerative diseases and dysfunctions. However, though previous studies have shown the presence of NADPH oxidase subunits in cell culture and mouse brain, they have not provided detailed high power resolution data. Therefore, we investigated the distributions of the p47Phox and gp91Phox subunits in rat brain using immunohistochemical approach. Cortex, hippocampus, and Purkinje cells of cerebellum were prominently stained by p47Phox and gp91Phox antibodies. As compared with the distributions of p47Phox, gp91Phox in mouse, some differences in the rat brain were observed in the hippocampus, thalamus, amygdala, reticular nucleus, and basal ganglia. Additionally, at the cellular level, most p47Phox immunoreactivity was largely confined to cell bodies and proximal portions of the dendritic tree. Taken together, the widespread observed distributions of p47Phox and gp91Phox subunits indicate that they are probably needed to maintain normal brain function. PMID- 15804440 TI - Insulin-like 6 immunoreactivity in the mouse brain and testis. AB - Insulin-like 6 immunoreactivity (irINSL6) was detected in Leydig cells of the mouse testis. In the brain, labeled somata were detected mainly in the caudal hypothalamus and midbrain. Double labeling the brainstem sections revealed that irINSL6 somata were 5-hydroxytryptamine (5-HT) positive. The presence of irINSL6 in discrete populations of hypothalamic and brainstem neurons and in Leydig cells of the testis suggests a diverse biological function of this novel peptide. PMID- 15804441 TI - Morphine decreases extracellular levels of glutamate in the anterior cingulate cortex: an in vivo microdialysis study in freely moving rats. AB - In the present study, we investigated the effect of morphine on the extracellular levels of glutamate in the anterior cingulate cortex (ACC) in freely moving rats using in vivo microdialysis coupled to high performance liquid chromatography and electrochemical detection. The results showed that either acute or chronic morphine treatment decreased the extracellular levels of glutamate in the ACC. Naloxone could reverse the decrease induced by chronic morphine treatment. The present study provided the first neurochemical evidence that morphine decreased extracellular levels of glutamate in the ACC, suggesting that glutamate in ACC is involved in the central actions of morphine. PMID- 15804442 TI - Vasoactive intestinal peptide in the amygdala inhibits tail flick reflexes in rats. AB - The present study was conducted to test the capability of a representative type of non-opioid peptides vasoactive intestinal peptide (VIP) in the amygdala to modulate nociception. Bilateral application of VIP into the basolateral region of the amygdala persistently suppressed radiant heat-evoked tail flick reflexes of anesthetized rats. The present result suggests that VIP synapses in the amygdala may play important roles in controlling pain, as with opioid synapses in the amygdala. This result also implies that local VIP in the amygdala is likely to subserve activating the descending antinociceptive systems of the brainstem from the amygdala. PMID- 15804443 TI - Production of IL-27 and other IL-12 family cytokines by microglia and their subpopulations. AB - Production of IL-27 and other IL-12 family cytokines by murine microglia were examined using RT-PCR, real-time RT-PCR and Western blot analysis. We show for the first time that murine microglia produce IL-27 in response to lipopolysaccharide (LPS) and/or interferon-gamma. Primary microglia, but not their cell lines, also induce IL-12 and IL-23 upon above stimulation. Therefore, microglia may play a critical role initiating Th1 responses via producing IL-12 family cytokines in the brain. PMID- 15804444 TI - Activation of p38 mitogen-activated protein kinase in the early and peak phases of autoimmune neuritis in rat sciatic nerves. AB - To examine the involvement of p38 mitogen-activated protein kinase (MAPK) in autoimmune disorders of the peripheral nerve system, we analyzed the phosphorylation of p38 MAPK protein in the sciatic nerves of Lewis rats with experimental autoimmune neuritis (EAN). Western blot analysis showed that phosphorylated p38 (p-p38) MAPK protein was significantly increased in the sciatic nerves of rats in the early and peak phases of EAN, and declined gradually thereafter. Immunohistochemistry showed that p-p38 MAPK levels were increased in the infiltrating inflammatory cells, including T cells and macrophages, as well as in blood vessels and some Schwann cells in EAN-affected sciatic nerves, as compared to the sciatic nerves of controls. Some inflammatory cells and a few Schwann cells were also positive for TUNEL reaction at the peak and recovery phases of EAN. In conclusion, we postulate that the phosphorylation of p38 MAPK is involved in the elimination of infiltrating inflammatory cells during the course of EAN and may possibly modulate recovery in autoimmune disorders of the peripheral nervous system. PMID- 15804445 TI - A neuromagnetic analysis of the mechanism for generating auditory evoked fields. AB - Averaged and non-averaged neuromagnetic responses to repetitive transients of sound were analyzed in the frequency and time domains. It was found that ongoing oscillations between 3 and 16 Hz are relevant to the generation of auditory evoked fields (AEF). First, phases of the oscillation were locked at around the timings of N100m, but randomly distributed from trial to trial at the other timings. Secondly, amplitudes of the oscillation were larger on average at around the timings of N100m than at the other timings, although the amplitudes fluctuated from trial to trial with almost the same standard deviations throughout the periods of observation. Thirdly, spatial distributions of the oscillatory activities were often reduced to an equivalent current dipole in the auditory cortex at around the timings of N100m, but seldom reduced at the other timings. An explanation of these results would be to suppose several oscillators within the cortex whose phases are locked at around the timings of N100m, but fluctuated randomly at the other timings. PMID- 15804446 TI - Diagnostic subgroups of developmental dyslexia have different deficits in neural processing of tones and phonemes. AB - The present study addressed auditory processing in 8-11-year-old children with developmental dyslexia by means of event-related brain potentials (ERP). Cortical sound reception was evaluated by recording N250 responses to syllables and tones and cortical sound discrimination by analyzing the mismatch negativity (MMN) to syllable and tone changes. We found that both cortical sound reception and sound discrimination were impaired in dyslexic children. The analysis of the data obtained from two dyslexic subgroups, Dyslexics-1 being impaired in non-word reading (or both non-word and frequent word reading) and Dyslexics-2 in frequent word reading but not in non-word reading, revealed that the MMN was specifically diminished in the latter group whereas it was normal-like in Dyslexics-1. However, no differences were found between these subgroups in sound reception as indicated by the responses elicited by the standard stimuli. These results show that different diagnostic subgroups of dyslexics have different patterns of auditory processing deficits as suggested by similarly impaired sound reception in both dyslexic groups and the sound-discrimination impairment specific to one of the groups. PMID- 15804447 TI - Attention and prepulse inhibition: the effects of task-relevant, irrelevant, and no-task conditions. AB - We investigated whether attentional modulation of prepulse inhibition (PPI) is due to increased protection of processing of attended lead stimuli, decreased protection of processing of ignored lead stimuli, or a combination of both processes. Task and no-task trials, pre-cued by red and blue dots on a computer screen, were randomly intermixed. College student participants were instructed to do a tone duration judgment task on trials preceded by one color (task condition) and to do nothing on trials preceded by the other color (no-task condition). On task condition trials participants were instructed to count the number of longer duration tones of a particular pitch (attended condition) and to ignore tones of a different pitch (ignored condition). White noise startle stimuli were presented at 60 ms and 120 ms lead intervals on some trials in each condition. Additional startle stimuli were presented during the inter-trial intervals to measure baseline (unmodified) startle response. PPI in the attended condition was reliably greater than that in both the ignored and no-task conditions. PPI did not differ between the ignored and no-task conditions. The results are consistent with the conclusion that attentional modulation of PPI is due to increased protection of attended stimuli and not to decreased protection of ignored stimuli. Possible reasons for robust attentional modulation at the 60 ms lead interval as well as the usual 120 ms lead interval are discussed. PMID- 15804448 TI - Extraversion and cortical activation during memory performance. AB - In this study we analyzed the influence of the personality dimension extraversion introversion (E) on the level and topographical distribution of cortical activation. In 62 participants (32 introverts and 30 extraverts), we measured the extent of Event-Related Desynchronization (ERD) in the EEG during performance of a short-term memory (i.e., temporary maintenance of information) and a more complex working memory task (i.e., temporary maintenance and active manipulation of information). The results indicate that during performance of both tasks, introverts display a larger amount of ERD than extraverted individuals. Moreover, the present E effects largely match previous studies as to the restriction of these effects to lower EEG frequency ranges (approx. 4-8 Hz). Topographical analyses show that the E effects are primarily present over (right-hemispheric) frontal and parietal regions of the cerebral cortex. PMID- 15804449 TI - Alpha-contingent EEG feedback reduces SPECT rCBF variability. AB - EEG feedback methods, which link the occurrence of alpha to the presentation of repeated visual stimuli, reduce the relative variability of subsequent, alpha blocking event durations. The temporal association between electro-cortical field activation and regional cerebral blood flow (rCBF) led us to investigate whether the reduced variability of alpha-blocking durations with feedback is associated with a reduction in rCBF variability. Reduced variability in the rCBF response domain under EEG feedback control might have methodological implications for future brain-imaging studies. Visual stimuli were randomly presented to seven subjects, contingent upon the occurrence of alpha (alpha-contingent stimulation (ACS)) or alpha-blocking (not-alpha-blocking-contingent stimulation (NACS)) events. We employed a within-subjects design. rCBF was measured from multiple, cortical and sub-cortical regions. The primary dependent variables were the Mean, Standard Deviation and the ratio of Mean/Standard Deviation of: 1) the alpha blocking response durations and 2) the temporally summated rCBF responses within the Visual Associative regions of interest (ROIs). Additional within-subjects rCBF measures were derived to quantify the variance-reducing effects of ACS across multiple, distributed areas of the brain. Both EEG and rCBF measures demonstrated decreased variability under ACS. This improved control was seen for localized as well as anatomically distributed rCBF measures. PMID- 15804450 TI - Pain perception, obstructive imagery and phase-ordered gamma oscillations. AB - The neural mechanisms underlying pain perception and anti-nociceptive effects of mental imagery are not well understood. Using a measure of phase-ordered beta and gamma EEG oscillations in response to painful electric stimulation, we recently found that somatosensory event-related phase-ordered gamma oscillations (38-42 Hz), elicited by the onset of painful stimuli over Cz scalp site, were linearly related to pain perception. In the present study, 38 subjects were engaged in a painful stimulus detection task using an oddball paradigm. This task was performed under a condition in which subjects were required simply to count the number of target stimuli (pain condition) and under another condition in which subjects were required to produce an obstructive mental imagery of painful stimulus perception (obstructive imagery). Only EEG responses to standard stimuli were analyzed in this study. Correlation analysis of sweeps for each individual revealed brief intervals of phase ordering of EEG patterns in the beta and gamma bands. The frequencies of interest were the beta1 (26-30 Hz), beta2 (30-34 Hz), gamma1 (34-38 Hz), gamma2 (38-42 Hz) and gamma3 (42-46 Hz) bands. Obstructive imagery treatment, compared to pain condition, significantly reduced pain perception. This reduction was paralleled by significant decreases of evoked phase-ordered gamma2 and gamma3 patterns over Cz scalp site. Phase-ordered oscillations at Cz scalp site, for both gamma2 and gamma3 bands, significantly predicted pain ratings during pain condition. Phase-ordered oscillation scores, obtained for these gamma bands over parietal and frontal scalp sites, resulted the best predictor of pain ratings during obstructive imagery. This study provides evidence for the role of gamma oscillations in the subjective experience of pain. Further, it has provided support for the view that pain reduction during obstructive mental imagery is the product of an inhibitory process involving frontal and parietal cortical regions. PMID- 15804451 TI - The influence of task demand and learning on the psychophysiological response. AB - The level of expertise of an operator may significantly influence his/her psychophysiological response to high task demand. A naive individual may invest considerable mental effort during performance of a difficult task and psychophysiological reactivity will be high compared to the psychophysiological response of a highly skilled operator. A study on multitasking performance was conducted to investigate the interaction between learning and task demand on psychophysiological reactivity. Thirty naive participants performed high and low demand versions of the Multi-attribute Task Battery (MATB) over a learning period of 64 min. High and low task demand setting were preset via a pilot study. Psychophysiological variables were collected from four channels of EEG (Cz, P3, P4, Pz), ECG, EOG and respiration rate to measure the impact of task demand and learning. Several variables were sensitive to the task demand manipulation but not time-on-task, e.g., heart rate, Theta activity at parietal sites. The sensitivity of certain variables to high demand was compromised by skill acquisition, e.g., respiration rate, suppression of alpha activity. A sustained learning effect was observed during the high demand condition only; multiple regression analyses revealed that specific psychophysiological variables predicted learning at different stages on the learning curve. The implications for the sensitivity of psychophysiological variables are discussed. PMID- 15804452 TI - The relationship of hostility, negative affect and ethnicity to cardiovascular responses: an ambulatory study in Singapore. AB - This study tested the hypotheses that ambulatory heart rate and blood pressure would be higher for individuals high but not low in hostility when they experienced negative affect or social stress and that this interaction would be stronger for Indians compared with other Singapore ethnic groups. Ambulatory blood pressure monitoring was done on 108 male Singapore patrol officers as they went about their daily duties. After each BP measurement participants completed a computerized questionnaire including items on emotional experience. Individuals high in hostility showed higher systolic blood pressure when reporting negative affect whereas this was not true for those low in hostility. Ethnic differences were obtained such that Indians showed an increase in mean arterial pressure when angered whereas MAP was negatively related to anger for Malays and unrelated for Chinese. Also a three-way interaction between ethnicity, hostility, and social stress indicated that hostility and social stress interacted in their effects on DBP for Indian participants but not for Chinese or Malays. Finally, a three-way interaction was obtained between ethnicity, hostility and negative affect for heart rate in which heart rate increased with increasing levels of negative affect for Chinese high in hostility and Malays low in hostility but decreased with increasing negative affect for all other participants. These data are consistent with higher CHD rates among individuals high in hostility and also provide additional evidence on ethnic differences in cardiovascular reactivity in Singapore. PMID- 15804453 TI - Protective effect of diphenyl diselenide on acute liver damage induced by 2 nitropropane in rats. AB - The effect of diphenyl diselenide, (PhSe)2, administration on 2-nitropropane (2 NP)-induced hepatic damage was examined in male rats. Rats were pre-treated with a single dose of diphenyl diselenide (10, 50 or 100 micromol/kg). Afterward, they received only one dose of 2-NP (100 mg/kg body weight dissolved in olive oil). The parameters that indicate tissue damage such as plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alpha-fetoprotein (AFP), creatinine and urea were determined. Since toxicity induced by 2-NP is related to oxidative stress, lipid peroxidation was also evaluated. Diphenyl diselenide (100 micromol/kg) significantly reduced plasma ALT, gamma-GGT, AFP levels when compared to 2-NP group. Treatment with diphenyl diselenide, at all doses, effectively protects the increase of lipid peroxidation when compared to 2-NP group. Histological examination revealed that 2-NP treatment causes a moderate swelling and degenerative alterations on hepatocytes and diphenyl diselenide (100 micromol/kg) protects against these alterations. Diphenyl diselenide (50 and 100 micromol/kg) significantly decreased the urea level. This study evidences the protective effect of diphenyl diselenide by 2-NP-induced acute hepatic damage. PMID- 15804454 TI - Effect of cytokine treatment on the neurogenesis process in the brain of soman poisoned mice. AB - We previously described that enhanced proliferation of neural progenitors occurred in the subgranular zone (SGZ) of the dentate gyrus and in the subventricular zone (SVZ) of the mouse brain following soman poisoning. Then, a discrete number of these cells seemed to migrate and engraft into the main damaged brain regions (hippocampus; septum and amygdala) and subsequently differentiate into neurons. In the present study, the effect of a cytokine treatment on the neurogenesis process was evaluated. For this purpose, subcutaneous injection of a cocktail of 40 microg/kg epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) was administered daily to soman poisoned mice (110 microg/kg soman and 5.0 mg/kg methyl nitrate atropine), from post-soman days 1 to 8. To label replicating neural progenitors, 200 mg/kg bromodeoxyuridine (BrdU) was injected twice a day between post-soman days 6 and 8. Mice were sacrificed on post-soman day 9 or 34. On post-soman day 9, the cytokine treatment had no effect on the proliferation of neural progenitors in the SVZ and SGZ, as assessed by BrdU immunochemistry. However, this treatment seemed to promote the migration of neural precursor cells from the proliferative areas towards damaged brain regions. Indeed, in the CA1 hippocampal layer of soman-poisoned mice, on post-soman day 34, the cytokine treatment increased the number of healthy pyramidal neurons stained by hemalun-eosin dye. The cytokine treatment also augmented the number of BrdU-labeled cells in the CA1 hippocampal layer and amygdala. Interestingly, the administration of cytokines resulted in the differentiation of BrdU-positive cells into new neurons in the CA1 hippocampal layer, whereas astrocytic differentiation was preferentially observed in the amygdala. PMID- 15804455 TI - Brain regional responses in antioxidant system to alpha-lipoic acid in arsenic intoxicated rat. AB - Impaired antioxidant defense mechanisms and oxidative stress are implicated in the pathogenesis of arsenic toxicity. Our study was designed to determine whether alpha-lipoic acid, which has been shown to have substantial antioxidant properties, when administered (70 mg/kg body weight) once daily for 60 days along with arsenic (100 ppm sodium arsenite mixed in drinking water) would prevent arsenic-induced changes in antioxidant defense system, superoxide dismutase (SOD total SOD, Mn SOD, Cu/Zn SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) in rat brain regions such as cortex, hypothalamus, striatum, cerebellum and hippocampus. The present study also examined the effect of alpha-lipoic acid over arsenic-induced oxidant production and lipid peroxidation level (LPO) in discrete brain regions of rats. The cortex, striatum and hippocampus showed greater decreases in GSH-Px enzyme activity than cerebellum and hypothalamus with arsenic exposure. Striatum had the greatest percentage of decreased activities of total SOD and Mn SOD, whereas cortex had the greatest percentage decrease in the activity of Cu/Zn SOD in arsenic-alone treated rats. Hypothalamus and cerebellum exhibited the lowest catalase activity among all tested regions in arsenic-only treated rats. Rate of dichlorofluorescin oxidation, an indication of reactive oxygen species and other intracellular oxidants production was increased with arsenic exposure in all brain regions studied. Cortex, hippocampus and striatum exhibited greater increase of LPO levels than cerebellum and hypothalamus. SOD, CAT, GSH-Px activities were upregulated in arsenic plus lipoic acid treated versus arsenic-only treated rats. Also, simultaneous lipoic acid treatment along with arsenic proved to be sufficient in reducing oxidant production and LPO level in all rat brain regions. Our results demonstrate that arsenic-induced deficits in antioxidant enzyme activities and increase in oxidant production and lipid peroxidation level in brain regions can be overcome through simultaneous treatment with lipoic acid. PMID- 15804456 TI - Cytotoxic effects in mammalian Vero cells exposed to pentachlorophenol. AB - The effects of pentachlorophenol have been studied on diverse biological systems both in vivo and in vitro, however the cellular basis of the pronounced cytotoxicity of this organochlorine compound is poorly understood. In this work, morphological and biochemical analyses were carried out to identify the primary targets of pentachlorophenol toxicity in mammalian cells. Our results show that pentachlorophenol is a very potent cytotoxic drug that displays an unusual and interesting mode of action in Vero cells. Although this compound is a powerful uncoupler of oxidative phosphorylation, we present the novel finding that lysosome destabilization is an early cytotoxic response that precedes the mitochondrial dysfunction. In addition, soon after exposure to moderate doses of pentachlorophenol, a significant number of cells initiate an apoptotic death process identified by the condensed and fragmented state of their nuclei. These results demonstrate that there are multiple potential targets of PCP-induced toxicity in mammalian cells, and the need to develop further experimental studies for the risk assessment of this environmental pollutant. PMID- 15804457 TI - Comparative study on two freshwater invertebrates for monitoring environmental lead exposure. AB - Two freshwater invertebrate organisms, Biomphalaria glabrata and Lumbriculus variegatus, were tested as potential experimental animal models to assess Pb exposure using acute laboratory bioassays. Since long, the enzyme delta aminolevulinic acid dehydratase (ALA-D) has been recognised as a useful biomarker of Pb exposure and effect. Therefore, determinations of ALA-D activity were performed in the whole body soft tissues of pigmented and non-pigmented gastropods B. glabrata and in the oligochaete L. variegatus. The organisms were exposed to varying concentrations of Pb for 48 h. Levels of Pb in the invertebrates were also analysed. Highly significant negative correlations were found between the enzymatic activity and the levels of Pb exposure, and also between the enzymatic activity and the metal incorporated by the invertebrates. No significant differences were found in the enzyme activity nor in the resulting metal accumulation based on gastropod pigmentation. The values of Pb concentration that produce 50% of inhibition on the enzyme activity (EIC50) were 0.023 and 0.029 mg Pb/L for pigmented and non-pigmented B. glabrata, respectively. A much higher value was found for L. variegatus (0.703 mg Pb/L). The non-observed effect concentration (NOEC) on enzyme activity for the oligochaetes was 0.05 mg Pb/L, about twice the EIC50 calculated for the gastropods. These data showed that both pigmented and non-pigmented B. glabata were much more sensitive organisms than the oligochaetes. The differences in enzyme inhibition could be attributed, at least partly, to differences in the metal body burden found between the organisms, since levels of Pb accumulated by B. glabrata were approximately three times higher compared to those observed in L. variegatus. Therefore B. glabrata showed to be a more suitable and reliable bioindicator organism for assessing Pb contamination in aquatic ecosystems, especially at low levels of metal exposure. PMID- 15804458 TI - Resveratrol, a polyphenolic phytoalexin protects against cyclosporine-induced nephrotoxicity through nitric oxide dependent mechanism. AB - Cyclosporine A (CsA) is a potent and effective immunosuppressive agent, but its use is frequently accompanied by severe renal toxicity. The causes for the nephrotoxicity of CsA have not been fully elucidated. Intrarenal vasoconstriction induced by several different mediators, both in humans and experimental animals have been proposed. The present study was designed to investigate the possible protective effect of resveratrol on CsA-induced nephrotoxicity and to explore the possible mechanism involved in resveratrol's effect. Eight groups of rats were employed in this study, group 1 served as control, group 2 rats were treated with olive oil (vehicle for CsA), group 3 rats were treated with CsA (20 mg/kg, s.c. for 21 days), groups 4, 5 and 6 received CsA along with resveratrol (2, 5 and 10 mg/kg, p.o. 24 h before and 21 days concurrently), respectively, group 7 rats were treated with NOS inhibitor, L-NAME (10 mg/kg) along with resveratrol and CsA and group 8 rats received L-NAME along with CsA. CsA administration for 21 days resulted in a marked renal oxidative stress, significantly deranged the renal functions, reduced the tissue and urine nitrite levels and markedly altered the renal morphology. Treatment with resveratrol (5 and 10 mg/kg) significantly improved the renal dysfunction; tissue and urine total nitric oxide levels, renal oxidative stress and prevented the alterations in renal morphology. Concurrent administration of L-NAME blocked the protective effect of resveratrol indicating that resveratrol exerts its protective effect by releasing nitric oxide. These results clearly demonstrate the pivotal role of nitric oxide in etiology of CsA nephrotoxicity and indicate the renoprotective potential of resveratrol in CsA nephrotoxicity. PMID- 15804459 TI - Evaluation of immuno- and reproductive toxicities and association between immunotoxicological and genotoxicological parameters in waste incineration workers. AB - In this study, we investigated immunotoxicity levels of 2,3,7,8 tetrachlorodibenzo-p-dioxin in 31 waste incineration workers and in 84 control subjects, and evaluated the association between immunological and genotoxicological parameters. DNA damage in mononuclear and polynuclear lymphocytes, and the level of the urinary metabolites, 1-OHP and 2-naphthol, were evaluated in both waste incineration workers and control subjects. Significant differences were detected in these values between exposed and control groups. Number of sperms was lower in the waste incineration workers than in the control subjects, as was the percentage of motile sperms, but a significant difference existed only in the number of sperms (p=0.05). DNA damage in the spermatozoa of waste incineration workers and control subjects measured 1.40+/-0.08 and 1.26+/ 0.03, respectively (p=0.001). Specific surface antigens of peripheral lymphocytes, namely, CD3, CD4, CD8, CD19, and CD69 were used to measure immune status in waste incineration workers and control subjects. There was no significant difference in T- and B-cell profiles showed between waste incineration workers and control subjects (p=0.684 and 0.157). In addition, the ratio of T helper cells to T cytotoxic cells was also not remarkably different between groups (p=0.174). However, T-cell activation was found to be significantly higher in the waste incineration workers than in the control subjects (p=0.001), although B-cell activation did not exhibit this trend. The levels of two cytokines (IL-4 an INF-gamma) and four immunoglobulins (IgA, IgE, IgG, and IgM) were also measured in the experimental population. All immunoglobulin types were found in lower amounts in the waste incineration workers, but this diaparity was not significant one. In addition, the levels of two cytokines, IL-4 and INF-gamma, were also found to be lower in the waste incineration workers than in the control subjects, and only in IL-4 was a significant difference determined to exist. PMID- 15804460 TI - T-2 toxin immunotoxicity on human B and T lymphoid cell lines. AB - T-2 toxin belongs to a group of mycotoxins synthesized by Fusarium fungi that are widely encountered as natural contaminants in cereals. Human lymphoid cell lines of T (MOLT-4) or B (IM-9) lineage were used to characterize the cytotoxic effects mediated by T-2 at different concentrations (0.1 pg/ml to 1 microg/ml). After 24 h, membrane damage was observed by Trypan blue dye exclusion in IM-9 cells with a 50% cytotoxic concentration (CC50) of 0.2 ng/ml, whereas CC50 for MOLT-4 cells was 0.6 microg/ml (gmicro). At a T-2 concentration of 0.01 microg/ml, apoptosis was seen in MOLT-4 cells by Annexin V binding as early as after 4 h. T-2 toxin determined sustained (48 h) immunosuppression on both cell lines, as evaluated by BrdU and MTT assays. Cytotoxicity appeared to be due to early apoptosis in MOLT-4 cells, as indicated by increased Annexin V binding and activation of caspase-3, and to direct cell membrane damage in IM-9 cells. PMID- 15804462 TI - A vignette from breast cancer history: one hundred years after the Greenough report. PMID- 15804463 TI - Prehospital care from Napoleon to Mars: the surgeon's role. PMID- 15804465 TI - Predictors of invasive breast cancer in patients with an initial diagnosis of ductal carcinoma in situ: a guide to selective use of sentinel lymph node biopsy in management of ductal carcinoma in situ. AB - BACKGROUND: The role of sentinel lymph node biopsy (SLNB) in patients with an initial diagnosis of ductal carcinoma in situ (DCIS) has not been well defined. The purpose of our study was to determine when the risk of finding invasive disease on final pathology in patients with an initial diagnosis of DCIS was sufficiently high to justify use of SLNB. STUDY DESIGN: The records of 398 consecutive patients from our prospective database with an initial diagnosis of DCIS, treated between July 1999 and December 2002, were analyzed. Associations between clinical and pathologic factors and patient selection for SLNB and outcomes were analyzed for significance using univariate and multivariate analyses. RESULTS: Of the 398 patients, 80 (20%) were found to have invasive disease on final pathology. Multivariate analysis revealed 4 independent predictors of invasive cancer on final pathology: 55 years of age or younger (odds ratio [OR], 2.19; p = 0.024), diagnosis by core-needle biopsy (OR, 3.76; p = 0.006), mammographic DCIS size of at least 4 cm (OR, 2.92; p = 0.001), and high grade DCIS (OR, 3.06; p = 0.002). A total of 141 patients (35%) underwent SLNB as a component of their initial operation. Multivariate analysis revealed that the presence of comedonecrosis (OR, 2.69; p = 0.007) and larger mammographic DCIS size (OR, 1.18; p = 0.0002) were independent predictors of patients' undergoing SLNB. Of these 141 patients, 103 (73%) were diagnosed by core-needle biopsy, 42 (30%) had invasive disease on final pathology, and 14 (10%) had a positive sentinel lymph node: 12 (86%) by hematoxylin and eosin staining and 2 by immunohistochemistry. The only independent predictor of a positive SLN was the presence of a palpable tumor (OR, 4.28, p = 0.042). Of these 14 patients with a positive sentinel node, only 11 (79%) had invasive cancer on final pathology. CONCLUSIONS: SLNB should not be performed routinely for all patients with an initial diagnosis of DCIS. Risks and benefits of SLNB should be discussed with patients who are younger, are diagnosed by core-needle biopsy, or have large or high-grade DCIS. PMID- 15804464 TI - Kidney transplantation under minimal immunosuppression after pretransplant lymphoid depletion with Thymoglobulin or Campath. AB - BACKGROUND: Multiple drug immunosuppression has allowed the near elimination of rejection, but without commensurate improvements in longterm graft survival and at the cost of quality of life. We have suggested that transplantation outcomes can be improved by modifying the timing and dosage of immunosuppression to facilitate natural mechanisms of alloengraftment and acquired tolerance. STUDY DESIGN: Two therapeutic principles were applied for kidney transplantation: pretransplant recipient conditioning with antilymphoid antibody preparations (Thymoglobulin [Sangstat] or Campath [ILEX Pharmaceuticals]), and minimal posttransplant immunosuppression with tacrolimus monotherapy including "spaced weaning" of maintenance doses when possible. The results in Thymoglobulin- (n = 101) and Campath-pretreated renal transplantation recipients (n = 90) were compared with those in 152 conventionally immunosuppressed recipients in the immediately preceding era. RESULTS: Spaced weaning was attempted in more than 90% of the kidney transplant recipients after pretreatment with both lymphoid depleting agents, and is currently in effect in two-thirds of the survivors. Although there was a much higher rate of acute rejection in the Thymoglobulin pretreated recipients than in either the Campath-pretreated or historic control recipients, patient and graft survival in both lymphoid depletion groups is at least equivalent to that of historic control patients. In the Thymoglobulin conditioned patients for whom followups are now 24 to 40 months, chronic allograft nephropathy (CAN) progressed at the same rate as in historic control patients. Selected patients on weaning developed donor-specific nonreactivity. CONCLUSIONS: After lymphoid depletion, kidney transplantation can be readily accomplished under minimal immunosuppression with less dependence on late maintenance immunosuppression and a better quality of life. Campath was the more effective agent for pretreatment. Guidelines for spaced weaning need additional refinement. PMID- 15804466 TI - Rates of reexcision for breast cancer after magnetic resonance imaging-guided bracket wire localization. AB - BACKGROUND: We performed this study to determine rates of close or transected cancer margins after magnetic resonance imaging-guided bracket wire localization for nonpalpable breast lesions. STUDY DESIGN: Of 243 women undergoing MRI-guided wire localizations, 26 had MRI bracket wire localization to excise either a known cancer (n = 19) or a suspicious MRI-detected lesion (n = 7). We reviewed patient age, preoperative diagnosis, operative intent, mammographic breast density, MRI lesion size, MRI enhancement curve and morphology, MRI Breast Imaging Reporting and Data System (BI-RADS) assessment code, number of bracket wires, and pathology size. We analyzed these findings for their relationship to obtaining clear margins at first operative excision. RESULTS: Twenty-one of 26 (81%) patients had cancer. Of 21 patients with cancer, 12 (57%) had negative margins at first excision and 9 (43%) had close/transected margins. MRI size > or = 4 cm was associated with a higher reexcision rate (7 of 9, 78%) than those < 4 cm (2 of 12, 17%) (p = 0.009). MRI BI-RADS score, enhancement curve, morphology, and preoperative core biopsy demonstrating ductal carcinoma in situ (DCIS) were not predictive of reexcision. The average number of wires used for bracketing increased with lesion size, but was not associated with improved outcomes. On pathology, cancer size was smaller in patients with negative margins (12 patients, 1.2 cm) than in those with close/transected margins (9 patients, 4.6 cm) (p < 0.001). Reexcision was based on close/transected margins involving DCIS alone (6, 67%), infiltrating ductal carcinoma and DCIS (2, 22%), or infiltrating ductal carcinoma alone (1, 11%). Reexcision pathology demonstrated DCIS (3, 33%), no residual cancer (5, 55%), and 1 patient was lost to followup (1, 11%). Interestingly, cancer patients who required reexcision were younger (p = 0.022), but breast density was not associated with reexcision. CONCLUSIONS: To our knowledge, this is the first report of MRI-guided bracket wire localization. Patients with MRI-detected lesions less than 4 cm had clear margins at first excision; larger MRI-detected lesions were more likely to have close/transected margins. Reexcision was often because of DCIS and was the only pathology found at reexcision, perhaps because MRI is more sensitive for detecting invasive carcinoma than DCIS. PMID- 15804467 TI - A randomized, controlled trial evaluating the impact of a computerized rounding and sign-out system on continuity of care and resident work hours. AB - BACKGROUND: Adoption of limits on resident work hours prompted us to develop a centralized, Web-based computerized rounding and sign-out system (UWCores) that securely stores sign-out information; automatically downloads patient data (vital signs, laboratories); and prints them to rounding, sign-out, and progress note templates. We tested the hypothesis that this tool would positively impact continuity of care and resident workflow by improving team communication involving patient handovers and streamlining inefficiencies, such as hand-copying patient data during work before rounds ("prerounds"). STUDY DESIGN: Fourteen inpatient resident teams (6 general surgery, 8 internal medicine) at two teaching hospitals participated in a 5-month, prospective, randomized, crossover study. Data collected included number of patients missed on resident rounds, subjective continuity of care quality and workflow efficiency with and without UWCores, and daily self-reported prerounding and rounding times and tasks. RESULTS: UWCores halved the number of patients missed on resident rounds (2.5 versus 5 patients/team/month, p = 0.0001); residents spent 40% more of their prerounds time seeing patients (p = 0.36); residents reported better sign-out quality (69.6% agree or strongly agree); and improved continuity of care (66.1% agree or strongly agree). UWCores halved the portion of prerounding time spent hand copying basic data (p < 0.0001); it shortened team rounds by 1.5 minutes/patient (p = 0.0006); and residents reported finishing their work sooner using UWCores (82.1% agree or strongly agree). CONCLUSIONS: This system enhances patient care by decreasing patients missed on resident rounds and improving resident-reported quality of sign-out and continuity of care. It decreases by up to 3 hours per week (range 1.5 to 3) the time used by residents to complete rounds; it diverts prerounding time from recopying data to more productive tasks; and it facilitates meeting the 80-hour work week requirement by helping residents finish their work sooner. PMID- 15804468 TI - Comparison of training on two laparoscopic simulators and assessment of skills transfer to surgical performance. AB - BACKGROUND: Several studies have investigated the transfer of surgical trainees' skills acquired on surgical simulators to the operating room setting. The purpose of this study was to compare the effectiveness of two laparoscopic surgery simulators by assessing the transfer of skills learned on simulators to closely matched surgical tasks in the animal laboratory. STUDY DESIGN: In this post-test only Control group study design, 46 surgically naive medical student volunteers were randomly assigned to one of three groups: Tower Trainer group (n = 16), LapSim group (n = 17), and Control group (n = 13). Outcomes measures included both time and accuracy scores on three laparoscopic tasks (Task 1: Grasp and Place; Task 2: Run the Bowel; Task 3: Clip and Cut) performed on live anesthetized pigs, and a global rating of overall performance as judged by four experienced surgeons. RESULTS: The Tower Trainer group performed significantly better than the Control group on 1 of 7 outcomes measures-Task 3: Time (p < 0.032), although the LapSim group performed significantly better than the Control group on 2 of 7 measures-Task 3: Time (p < 0.008) and Global score (p < 0.005). In comparing the two simulators, the LapSim group performed significantly better than the Tower Trainer group on 3 of 7 outcomes measures-Task 2: Time (p < 0.032), Task 2: Accuracy (p < 0.030) and Global score (p < 0.005), although the Tower Trainer group did not perform significantly better than the LapSim group on any measure. CONCLUSIONS: This study demonstrated that naive subjects trained on a virtual-reality part-task trainer performed better on live surgical tasks in a porcine model as compared with those trained with a traditional box trainer. These findings could aid in selection of appropriate training methodologies. PMID- 15804469 TI - Surgical transplant physical examination: correlation of renal resistance index and biopsy-proven chronic allograft nephropathy. AB - BACKGROUND: Chronic allograft nephropathy (CAN) remains the leading cause of late renal allograft loss that is minimally responsive to therapy once graft dysfunction is clinically evident. A screening test capable of identifying individuals at high risk for CAN would be a valuable adjunct to patient care, but to be cost effective, should be administered during routine evaluations by transplantation clinicians. STUDY DESIGN: We have compared the resistive index (RI) as measured by Doppler ultrasonography with subsequent biopsy findings on 91 renal allograft recipients who had a subsequent protocol-directed biopsy at least 3 months after renal transplant. All ultrasonography was performed by the transplantation surgical staff without involving the radiology department or a separate appointment time. RESULTS: Twenty-one patients had RI >/= 80 (average 621 days posttransplantation). Among these individuals, the subsequent incidence of CAN was 38%. Length of time between initial assessment of increased RI and biopsy-proved CAN averaged 233 days. The remaining 70 patients with RI < 80 had an incidence of CAN of 11.4% (p = 0.018). There were minimal complications from these biopsies. Sensitivity and specificity of an elevated RI in predicting CAN were 50% and 83%, respectively. The negative predicted value of an elevated RI in determination of CAN was 89%. CONCLUSIONS: These results suggest that elevated RI is an early predictor of histologically relevant CAN, possibly a result of burgeoning vasculopathy. The technical expertise required to make this appraisal is well within the capabilities of transplantation surgeons and trainees. Early evidence of CAN may allow for a targeted change in therapy before clinically significant injury. Ultrasonography should become a routine part of a transplantation clinic evaluation. PMID- 15804470 TI - Placental umbilical cord whole blood transfusion: a safe and genuine blood substitute for patients of the under-resourced world at emergency. AB - BACKGROUND: Afterbirth or placenta is normally discarded. But placental cord blood, because of its rich mix of fetal and adult hemoglobin, plasma, and high platelet count has the potential to be a safe alternative to adult blood. STUDY DESIGN: We transfused 413 U (range 50 mL to 146 mL; mean 86 mL+/-7.6 mL SD; median 80 mL; mean packed cell volume 48+/-4.1% SD; mean hemoglobin concentration 16.2 g/dL+/-1.8 g/dL SD) of placental umbilical cord whole blood, after lower uterine cesarean section from consenting mothers, to 129 informed consenting patients, after screening by the institutional ethics committee, from April 1, 1999, until the present. RESULTS: The list of consenting patients included 54 men and 75 women. Patient age varied from 2 years to 86 years. Seventy-three patients (56.58%) suffered from advanced cancer and 56 (43.42%) patients had other diseases like ankylosing spondylitis, lupus erythematosus, rheumatoid arthritis, aplastic anemia, and thalassemia major. We have not encountered a single case of immunologic or nonimmunologic reaction so far. CONCLUSIONS: We suggest that medical institutions could use this gift of nature, which is free from infection, hypoantigenic with altered metabolic profile, filled with growth factor and cytokine-filled plasma, with the potential of higher oxygen-carrying capacity than adult blood, as an emergency source of blood for the management of disaster or crises anywhere in the world. It also may be safely used for the treatment of malignant and nonmalignant disorders. PMID- 15804471 TI - Duration of increased bleeding tendency after cessation of aspirin therapy. AB - BACKGROUND: Aspirin has a significant effect on hemostasis, so it is often recommended that patients taking aspirin discontinue treatment before elective surgery. While off aspirin, these patients may be at risk of thrombosis. The optimum period of time that aspirin should be withheld is controversial. The aim of this study was to establish the duration of the antihemostatic effect of prolonged aspirin therapy. STUDY DESIGN: In a prospective study, 51 healthy volunteers were randomly assigned into 3 groups, each receiving an identical tablet for 14 days. One group received a placebo tablet; individuals in the other two groups received either 75 mg or 300 mg of aspirin once a day. Template bleeding times and specific platelet function testing (using the PFA-100; Dade Behring) were carried out on subjects before therapy and again after its completion until they returned to baseline. RESULTS: Thirty-eight volunteers complied sufficiently with the protocol to provide useful results. All bleeding times normalized within 96 hours and all platelet function tests within 144 hours after stopping aspirin. There was no demonstrable hemostatic defect in any volunteer persisting by or beyond the sixth day after treatment cessation. There was no apparent difference in duration of effect between those taking either 75 mg or 300 mg of aspirin. CONCLUSIONS: This study uses sensitive measures of platelet function to demonstrate the duration of increased bleeding tendency after withdrawal of aspirin therapy. It supports discontinuation of aspirin therapy 5 days before elective surgery (with the operation being performed on the sixth day). PMID- 15804472 TI - Inhibition of nitric oxide synthase reverses the effect of albumin on lung damage in burn. AB - BACKGROUND: Early colloid resuscitation in major burn patients has been stopped because of its deteriorating effect on thermal injury-induced vascular hyperpermeability. We hypothesized that inhibition of inducible nitric oxide synthase (iNOS) to stabilize endothelial permeability and to retain colloid solution in the vascular space will reverse its effect on lung damage. STUDY DESIGN: In experiment 1, specific pathogen free rats underwent 35% total-body surface area burn or sham burn and were given equal volumes (7.5 mL/kg) of normal saline or albumin from femoral veins for fluid resuscitation immediately after burn. In experiment 2, S-methylisothiourea (SMT, 7.5 mg/kg, IP) was given immediately after burn to rats from different groups, as in experiment 1. At 8 hours after burn, blood was assayed for peroxynitrite-mediated dihydrorhodamine 123 (DHR 123) oxidation, and lung tissues were harvested for myeloperoxidase (MPO) determination and histologic studies. Pulmonary microvascular dysfunction was quantified by measuring the extravasations of Evans blue dye. RESULTS: Blood peroxynitrite level and iNOS expression, MPO activity, permeability, and inflammatory cell infiltration of lungs were significantly induced after thermal injury. Albumin resuscitation after burn without iNOS inhibition enhanced thermal injury-induced lung damage with 10%, 14%, and 5% increases in blood DHR oxidation level, lung MPO activity, and lung permeability, respectively, compared with saline injection. In contrast, burn + SMT rats with albumin injection showed significant, 23%, 37%, and 20%, decreases, respectively, in blood DHR 123 oxidation level, lung MPO activity, and lung permeability compared with burn + SMT + saline rats. CONCLUSIONS: Thermal injury induced lung damage. Restoration of extracellular fluid in early burn shock with albumin markedly augmented the lung neutrophil deposition, lung permeability increase, and blood peroxynitrite level. Inhibition of iNOS before albumin supplementation reversed its damaging effects on thermal injury-induced lung dysfunction to beneficial ones. PMID- 15804473 TI - Physiologic trauma triage criteria in adult trauma patients: are they effective in saving lives by transporting patients to trauma centers? AB - BACKGROUND: Trauma triage criteria have been in place for many years and were updated in 1999 by the American College of Surgeons. We are unaware of any studies that have directly examined the ability of these criteria to reduce short term mortality by transporting patients to trauma centers rather than to noncenters. STUDY DESIGN: Retrospective observational cohort study of adult patients meeting physiologic triage criteria who were transported to 9 regional (Level I) trauma centers, 21 area (Level II) trauma centers, and 119 noncenters in New York in 1996 to 1998. For each triage criterion and for one or more of the criteria, odds ratios and their confidence intervals for mortality in regional and area trauma centers versus noncenters and odds ratios and their confidence intervals for mortality in regional centers versus area centers and noncenters were used to measure performance. RESULTS: Patients in regional trauma centers had considerably lower mortality than patients in area trauma centers and noncenters for two individual triage criteria and for patients with one or more triage criteria (odds ratio, 0.75; 95% CI, 0.63-0.90 for one or more criteria). Also, patients with head injuries who were treated in regional centers had notably lower mortality than patients in other hospitals (odds ratio, 0.67; 95% CI, 0.53-0.85). CONCLUSIONS: In New York, regional trauma centers exhibit considerably lower mortality than area trauma centers or noncenters for adult patients meeting specific physiologic triage criteria. It is important that population-based trauma systems with data from centers and noncenters be developed for the purpose of evaluating and redesigning trauma systems. PMID- 15804474 TI - Bariatric surgery for morbid obesity: health implications for patients, health professionals, and third-party payers. PMID- 15804475 TI - Challenges and a new vision for academic surgery. PMID- 15804476 TI - What's new in general surgery: burns and metabolism. PMID- 15804477 TI - Is laparoscopic inguinal hernia repair an operation of the past? PMID- 15804478 TI - Rationale for laparoscopic gastric bypass. PMID- 15804479 TI - Bleeding Meckel's diverticulum. PMID- 15804480 TI - Cervical spine fracture in the ankylosing spondylitis patient. PMID- 15804482 TI - Retrievable inferior vena cava filters in high-risk trauma patients. PMID- 15804483 TI - Should I lie about my work hours this week? PMID- 15804484 TI - Photodynamic therapy for neovascular age-related macular degeneration. PMID- 15804485 TI - Mandatory resident work hour limitations. PMID- 15804486 TI - Vascular malformations. PMID- 15804487 TI - Identification of a novel Wee1 isoform. AB - We have identified a novel isoform of Wee1 kinase (Wee1i), which uses Met215 of Wee1 as its initiation codon. RT-PCR, Western blot, and in situ hybridization verified wee1i expression in mammalian cells, rat brain, and rat thymus. Recombinant and partially purified Wee1i from rat thymus displayed kinase activity comparable to or higher than Wee1. The N-terminal 214 residues of Wee1 facilitate its ubiquitin-dependent degradation to trigger mitotic entry. Since Wee1i, lack of these 214 residues, it may evade this degradation and thus provide constitutive Wee1-like kinase activity to inhibit mitotic cell proliferation. Thus, Wee1i may play an important role in differentiation and in tumor suppression. PMID- 15804488 TI - Isolation and spectroscopic characterization of the membrane-bound nitrate reductase from Pseudomonas chlororaphis DSM 50135. AB - A nitrate reductase was solubilized with Triton X-100 from the membranes of Pseudomonas chlororaphis DSM 50135 grown microaerobically in the presence of nitrate. Like other membrane-bound nitrate reductases, it contains three subunits, of 129, 66 (64) and 24 kDa, referred to in the literature as alpha, beta and gamma, respectively. Electrocatalytic studies revealed that only the membrane-bound, not the solubilized form of the enzyme, can accept electrons from a menaquinone analog, menadione, whereas both forms can accept electrons from methylviologen. The isolated enzyme possesses several iron-sulfur clusters and a molybdopterin guanine dinucleotide active center. The iron-sulfur clusters can be grouped in two classes according to their redox properties, the high-potential and low-potential clusters. In the as-isolated enzyme, two forms of the molybdenum center, high- and low-pH, are detectable by electron paramagnetic resonance spectroscopy. The low-pH form shows a hyperfine splitting due to a proton, suggesting the presence of an -OHx ligand. Dithionite reduces the Mo(V) center to Mo(IV) and subsequent reoxidization with nitrate originates a new Mo(V) signal, identical to the oxidized low-pH form but lacking its characteristic hyperfine splitting. The isolated preparation also contains heme c (in a sub stoichiometric amount) with the ability to relay electrons to the molybdenum center, suggesting that this nitrate reductase may contain heme c instead of the heme b usually found in this class of enzymes. PMID- 15804489 TI - Regulation of the immune response to cestode infection by progesterone is due to its metabolism to estradiol. AB - The aim of this work was to investigate the role of progesterone during Taenia crassiceps cysticercosis, and the immunological mechanisms involved in its effects, by relating progesterone treatment to whole parasite counts, to host humoral and cellular immune response, to the presence or absence of nuclear receptors to sex steroids in splenocytes, and to serum sex steroid levels in infected mice of both genders. Progesterone treatment increased parasite loads two-fold in females and three-fold in males compared with control mice. The expression of the Th2 cytokine profile (IL-4, IL-6 and IL-10) was markedly increased in infected mice of both genders, while progesterone treatment returned this expression to basal levels. However, the Th1 cytokine profile (IFN-gamma and TNF-alpha) was not affected by infection, whilst progesterone treatment increased the expression of both cytokines two-fold compared to uninfected, infected and placebo-treated mice. Testosterone serum levels decreased in infected male mice by 95%, and treatment with progesterone did not affect them. In females, no change in testosterone levels was observed. Progesterone levels increased three fold only in progesterone-treated infected mice of both sexes, while estradiol levels in female and male progesterone-treated infected mice increased two-fold compared to infected control mice. The infection markedly induced the expression of progesterone receptor (PR) isoforms A and B in splenocytes of infected mice of both genders (five-fold). Metabolism of progesterone to estradiol was demonstrated by the use of the anti-estrogen tamoxifen, which reduced parasite loads 100% in infected mice of both sexes treated with progesterone. These results suggest that progesterone, possibly through its metabolism to estradiol, affects establishment, growth and reproduction of the helminth parasite T. crassiceps. PMID- 15804490 TI - Mutation in mce operons attenuates Mycobacterium tuberculosis virulence. AB - On the Mycobacterium tuberculosis genome there are four mce operons, all of which are similar in sequence and organization, and code for putatively exported proteins. To investigate whether Mce proteins are essential for virulence, we generated knock-out mutants in mce1, mce2 and mce3 operons of M. tuberculosis and evaluated their ability to multiply in a mammalian host. The allelic replacement was confirmed in each mutant strain by Southern blotting. RT-PCR experiments demonstrated the lack of in vitro expression of mutated genes in Deltamce1 and Deltamce2 mutants. On the other hand, no expression of mce3 was detected in either the wild-type or mutant strains. Similar doubling time and growth characteristics in in vitro culture were observed for mutants and parental strains. The intratracheal route was used to infect BALB/c mice with the Deltamce3, Deltamce2 and Deltamce1 mutants. Ten weeks after infection, all mice infected with the Deltamce mutants survived, while those infected with the wild type strain died. This long survival correlated with very low counts of colony forming units (CFU) in the lungs. Deltamce1-infected mice developed very few and small granulomas, while animals infected with Deltamce3 or Deltamce2 mutants showed delayed granuloma formation. Mice infected with Deltamce1 did not develop pneumonia, while animals infected with Deltamce3 and Deltamce2 mutants showed small pneumonic patches. In spleens, bacterial counts of mutant strains were less reduced than in lungs, compared with those of wild-type. In contrast, no such attenuation was observed when the intraperitoneal route was used for infection. Moreover, Deltamce1 mutants appear to be more virulent in lungs after intraperitoneal inoculation. In conclusion, mce operons seem to affect the virulence of M. tuberculosis in mice, depending on the route of infection. Hypotheses are discussed to explain this last issue. Thus, mutants in these genes seem to be good candidates for vaccine testing. PMID- 15804491 TI - Homing and memory patterns of human gammadelta T cells in physiopathological situations. AB - Vgamma9Vdelta2 are a heterogeneous population of T cells and comprise distinct naive, memory and effector populations that can be distinguished on the basis of surface marker expression and effector functions. We review here these recently studied features of Vgamma9Vdelta2 T lymphocyte biology and the roles they play in infectious and autoimmune diseases. PMID- 15804492 TI - Multiple melanocortin receptors are expressed in bone cells. AB - Melanocortin receptors belong to the seven transmembrane domain, G-protein coupled family of receptors. There are five members of this receptor family labeled MC1R-MC5R. These receptors are activated by fragments derived from a larger molecule, proopiomelanocortin (POMC) and include ACTH, alpha beta and gamma-MSH and beta-endorphin. Because of in vitro and in vivo data suggesting direct effects of these POMC molecules on bone and bone turnover, we examined bone and bone derived cells for the presence of the various members of the melanocortin receptor family. We report that the five known melanocortin receptors are expressed to varying degrees in osteoblast-like and osteoclastic cells. POMC fragments increased proliferation and expression of a variety of genes in osteoblastic cells. Furthermore, POMC mRNA was detected in osteoclastic cells. These data demonstrate that POMC-derived peptide hormones acting through high affinity melanocortin receptors have specific effects on bone cells. Thus, in addition to the indirect effects of POMC-derived hormones on bone turnover through their modulation of steroid hormone secretion, POMC fragments may have direct and specific effects on bone cell subpopulations. PMID- 15804493 TI - Biochemical markers of bone turnover are influenced by recently sustained fracture. AB - In striving to refine the clinical utility of different markers of bone metabolism, we should take into account numerous confounders, many of which are well known, such as sampling time, fasting status, and bone density. One further confounder may be ongoing fracture healing and/or post-fracture immobilization, which at least theoretically should impose an increased bone formation and resorption. Since both recent fracture and high bone turnover are independent predictors for new fracture, we thought it of importance to define the potential influence of such fracture on markers of bone turnover. From a population-based cohort of 1604 women, all 75 years old (the OPRA-study), 1024 women attended a clinical examination. The bone metabolism was assessed in serum, by three markers of bone formation [bone-specific alkaline phosphatase (S-Bone ALP), intact and N Mid osteocalcin (S-Total OC), and total carboxylated osteocalcin (S-cOC)], two markers of bone resorption [C-terminal cross-linked telopeptides of type I collagen (S-CTX) and tartrate-resistant acid phosphatase type 5b (S-TRACP5b)], and in urine by one marker of bone resorption [deoxypyridinoline/creatinine (U DPD/crea)] and two putative markers of bone resorption [urinary osteocalcins (U OC/crea)]. Current physical activity and retrospective fracture data were recorded by questionnaires. The fracture data, for the entire cohort of 1604 women, were validated with radiographic referrals and reports, saved since the beginning of the last century. All data provided, except date of occurrence of retrospectively sustained fracture, were thus obtained cross-sectionally and in all women at the age of 75. Fracture had ever been sustained by 727 of the entire cohort (n = 1604), and by 523 of the attending women (n = 1024). All markers were marginally higher (significant only for U-DPD/crea, P = 0.027) in women who had ever sustained fracture, compared to women without fracture. In women with recent retrospective fracture (since 2 years) (n = 100), the levels of all markers, except the two S-OCs, were significantly higher (r = 0.20-0.33, P = 0.049-0.001) the more recently the fracture had been sustained. Women with low current physical activity had elevated levels of U-DPD/crea (P < 0.001) and one U-OC (P = 0.014), while the other markers were unaffected. PMID- 15804494 TI - The Pre-Optic Anterior Hypothalamus (POAH) partially mediates the hypothermic response to hemorrhage in rats. AB - Two sets of experiments were performed to characterize the role of the Pre-Optic Area of the Anterior Hypothalamus (POAH) in the decrease in set point and hypothermia that follows severe hemorrhage. In the first set, lidocaine or artificial cerebrospinal fluid (ACSF) was microinjected into the POAH of rats at the time of hemorrhage. Lidocaine microinjection attenuated the hemorrhagic hypothermia by approximately 50%. The mean drop in core temperature (Tc) following hemorrhage was 1.5 degrees C with ACSF microinjection (N = 6), 0.70 degrees C (N = 6) with lidocaine, and 1.77 degrees C (N = 6) after sham microinjection. This partial attenuation of the hemorrhagic hypothermic response indicates that an intact POAH is necessary for at least some of the hypothermia following hemorrhage. In the second experimental set, hypothalamic tissue temperature (Thyp) was modulated in an attempt to alter the hemorrhagic hypothermic response. Bilateral closed-ended cannulas were inserted into the POAH. One cannula consisted of a water-perfused thermode to change local tissue temperature. The other housed a thermocouple to measure local temperature. The effectiveness of the thermode was first confirmed in conscious rats, evidenced by an inverse deflection in Tc upon Thyp modulation. Then, the POAH region was either heated, cooled, or sham perfused following hemorrhage. The mean drop in Tc following hemorrhage was 2.16 degrees C (N = 5) with hypothalamic heating, 1.35 degrees C (N = 5) with cooling, and 1.44 degrees C (N = 5) following the sham perfusion control. Heating of the POAH significantly exacerbated the hemorrhagic hypothermic response. These data further suggest that the POAH is at least partially responsible for mediating hemorrhagic hypothermia. PMID- 15804495 TI - Influence of the paraventricular nucleus and oxytocin on the retrograde stain of pubococcygeus muscle motoneurons in male rats. AB - Lumbosacral cord motoneurons innervating the pubococcygeus muscle (Pcm) at the pelvic floor of male rats were analyzed. We showed previously that these motoneurons participate in sexual functions and are sensitive to fluctuations of systemic androgen and estrogen. Though estrogen receptors have not been identified in Lamina IX at these spinal areas, the release of oxytocin from the paraventricular nucleus of the hypothalamus (PvN) has been found to control pelvic sexual physiology. We therefore worked on the hypothesis that steroid hormones in the PvN induce the release of oxytocin at the lumbosacral level to modulate the function of Pcm motoneurons. Four experiments were developed, and results were observed with the retrograde staining of motoneurons with horseradish peroxidase. Data indicated that morphometric parameters of Pcm motoneurons were significantly reduced after castration or blocking of the steroids at the PvN site, or following complete transection of the spinal cord at the T8 level. In each case, the reduction of the stain was recovered after intrathecal treatment with oxytocin. Thus, present results show that Pcm motoneurons respond to spinal oxytocin. The conclusive model that we propose is that steroids stimulate the PvN, causing the nucleus to release oxytocin at the level of the lumbosacral spinal cord, and the release of the peptide regulates the spread of the stain of Pcm motoneurons. This work also shows that motoneurons distal to a transected area in the spinal cord could respond to exogenous oxytocin, an important finding for the research of spinal cord lesioned subjects. PMID- 15804496 TI - More tryptophan hydroxylase in the brainstem dorsal raphe nucleus in depressed suicides. AB - Deficient serotonin neurotransmission in suicide is indicated by reduced brainstem serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA), fewer 5 HT(1A) autoreceptors and reduced cortical serotonin transporter binding in suicide victims. Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in the synthesis of 5-HT, and alterations in TPH could explain some of these findings. We sought to determine the amount of TPH immunoreactivity (TPH-IR) in the dorsal (DRN) and median (MRN) raphe nuclei in suicides and controls. Brainstems of suicide victims and controls (n = 11 pairs) were collected at autopsy, matched for age, sex and postmortem interval, frozen and sectioned (20 microm). Immunoautoradiography, using an antibody to label TPH, was performed, slides exposed to film and autoradiograms quantified by a computer-based image analysis system. We examined sections every 1000 microm throughout the whole length of the nucleus, performing statistical analysis only on those subjects for whom the raphe was complete (n = 8 pairs). TPH-IR (microCi/g) was higher in suicides than controls (S: 300.8 +/- 70.8 vs. C: 259.6 +/- 40.7, t = 2.57, df = 7, P = 0.04) in the dorsal raphe nucleus (DRN), and not different between suicides and controls (S: 251.3 +/- 44.2 vs. C: 235.9 +/- 27.4, t = 1.49, df = 7, P = 0.18) in the MRN. DRN TPH-IR was higher in male suicide victims (MS) compared to male controls (MC; MS: 318.4 +/- 54.4 vs. MC: 271.9 +/- 22.5, t = 2.66, df = 6, P = 0.03). The analysis of TPH-IR area and density at each DRN rostrocaudal levels showed higher area and density in suicides compared to controls in the rostral DRN and lower area and density in the caudal DRN. TPH-IR, an index of the amount of TPH enzyme, in the DRN is higher in depressed suicides. More TPH may be an upregulatory homeostatic response to impaired serotonin release or less autoreceptor activation. Alternatively, the serotonin impairment in suicide may be due to hypofunctional serotonin-synthesizing enzyme. PMID- 15804497 TI - Nitrergic innervation of trigeminal and hypoglossal motoneurons in the cat. AB - The present study was undertaken to determine the location of trigeminal and hypoglossal premotor neurons that express neuronal nitric oxide synthase (nNOS) in the cat. Cholera toxin subunit b (CTb) was injected into the trigeminal (mV) or the hypoglossal (mXII) motor nuclei in order to label the corresponding premotor neurons. CTb immunocytochemistry was combined with NADPH-d histochemistry or nNOS immunocytochemistry to identify premotor nitrergic (NADPH d(+)/CTb(+) or nNOS(+)/ CTb(+) double-labeled) neurons. Premotor trigeminal as well as premotor hypoglossal neurons were located in the ventro-medial medullary reticular formation in a region corresponding to the nucleus magnocellularis (Mc) and the ventral aspect of the nucleus reticularis gigantocellularis (NRGc). Following the injection of CTb into the mV, this region was found to contain a total of 60 +/- 15 double-labeled neurons on the ipsilateral side and 33 +/- 14 on the contralateral side. CTb injections into the mXII resulted in 40 +/- 17 double-labeled neurons in this region on the ipsilateral side and 16 +/- 5 on the contralateral side. Thus, we conclude that premotor trigeminal and premotor hypoglossal nitrergic cells coexist in the same medullary region. They are colocalized with a larger population of nitrergic cells (7200 +/- 23). Premotor neurons in other locations did not express nNOS. The present data demonstrate that a population of neurons within the Mc and the NRGc are the source of the nitrergic innervation of trigeminal and hypoglossal motoneurons. Based on the characteristics of nitric oxide actions and its diffusibility, we postulate that these neurons may serve to synchronize the activity of mV and mXII motoneurons. PMID- 15804498 TI - Glutamate uptake is attenuated in spinal deep dorsal and ventral horn in the rat spinal nerve ligation model. AB - Alteration of glutamatergic (GLU) neurotransmission within the spinal cord contributes to hyperalgesic and allodynic responses following nerve injury. In particular, changes in expression and efficacy of glutamate transporters have been reported. Excitatory, pain transmitting primary afferent neurons utilizing glutamate as an excitatory neurotransmitter project to both superficial (I-II) and deep (III-V) laminae of the dorsal horn. These experiments were designed to examine changes in glutamate uptake occurring concomitantly within the spinal deep dorsal and ventral horn in situ after experimentally induced neuropathic pain. In vivo voltammetry, using microelectrode arrays configured for enzyme based detection of GLU were employed. Sprague-Dawley rats had either sham surgery or tight ligation of L5 and L6 spinal nerves (SNL). Four to six weeks later, the L4-L6 spinal cord of chloral hydrate-anesthetized animals was exposed, and ceramic-based glutamate microelectrodes equipped with glass micropipettes 50 microm from the recording surfaces were placed stereotaxically at sites within the spinal cord. Pressure ejection of GLU into the ipsilateral L5-L6 spinal cord resulted in a 72% reduction of GLU uptake in SNL rats compared to sham controls in the ipsilateral L5-L6 deep dorsal horn and a 96% reduction in the ventral horn. In contrast, in the same animals, the contralateral L5-L6 or the ipsilateral L4 spinal cord showed no change in glutamate uptake. The data suggest that spinal nerve ligation produced attenuated glutamate uptake activity extending into the deep dorsal and ventral horn. The study suggests that plasticity related to spinal nerve injury produces widespread alteration in glutamate transporter function that may contribute to the pathophysiology of neuropathic pain. PMID- 15804499 TI - Effects of 3,4-methylenedioxymethamphetamine (MDMA, 'Ecstasy') and para methoxyamphetamine on striatal 5-HT when co-administered with moclobemide. AB - 3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") and para-methoxyamphetamine (PMA) are commonly used recreational drugs. PMA, often mistaken for MDMA, is reported to be more toxic in human use than MDMA. Both of these drugs have been shown to facilitate the release and prevent the reuptake of 5-hydroxytryptamine (5-HT, serotonin). PMA is also a potent inhibitor of monoamine oxidase type A (MAO-A), an enzyme responsible for the catabolism of 5-HT, and this characteristic may contribute to its increased toxicity. In humans, co administration of MDMA with the reversible MAO-A inhibitor moclobemide has led to increased apparent toxicity with ensuing fatalities. In the present study, using microdialysis, we examined the effects of co-administration of MDMA and PMA with moclobemide on extracellular concentrations of 5-HT and 5-hydroxy indol acetic acid (5-HIAA) in the striatum of the rat. 5-HT-mediated effects on body temperature and behavior were also recorded. Rats were pretreated with saline or 20 mg/kg (i.p.) moclobemide and 60 min later injected with 10 mg/kg MDMA, PMA, or saline. Dialysate samples were collected every 30 min for 5 h and analyzed by HPLC-ED. Both MDMA and PMA produced significant increases in extracellular 5-HT concentrations (590% and 360%, respectively, P < 0.05). Rats treated with PMA and MDMA displayed significantly increased 5-HT-related behaviors (P < 0.05). Furthermore, only MDMA was capable of producing additional significant increases in 5-HT concentrations (980%, P < 0.05) when co-administered with moclobemide. These data suggest that co-administration of MDMA with moclobemide increases extracellular 5-HT and 5-HT-mediated behaviors and may cause increased 5-HT related toxicity similar to that reported with PMA. PMID- 15804500 TI - Auditory compensation in myopic humans: involvement of binaural, monaural, or echo cues? AB - The purpose of this paper is to investigate the involvement of various auditory localization mechanisms in the improvement of auditory spatial sensitivity in the case of partial visual deprivation. We compared sensitivity to binaural, monaural, and echo cues between normal-sighted and myopic subjects. In an auditory task, which depended solely upon binaural processing, myopic and normal sighted subjects showed an almost equal localization performance. We also found that myopic subjects were no more disturbed than normal-sighted subjects when spectral shape cues were removed. On the other hand, near-sighted subjects exhibited better echolocation skills and a higher sensitivity to echo cues than normal-sighted subjects. We can therefore conclude that an improved auditory spatial sensitivity in myopic subjects may result from the increased use of echo information. PMID- 15804501 TI - Induction of long-term potentiation in single nociceptive dorsal horn neurons is blocked by the CaMKII inhibitor AIP. AB - Neuronal events leading to development of long-term potentiation (LTP) in the nociceptive pathways may be a cellular mechanism underlying central hyperalgesia. Here, we examine whether induction of LTP in nociceptive dorsal horn neurons at depths of 80-500 microm from the cord surface can be affected by spinal application of the Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) inhibitor AIP. Extracellular recordings from single neurons in intact urethane anesthetized Sprague-Dawley rats were performed, and the neuronal A-fiber and C fiber responses after sciatic nerve test pulses were defined according to latencies. A clear LTP of the nociceptive transmission following sciatic nerve high-frequency stimulation (HFS) was observed in single neurons in laminae I-IV of the dorsal horn. The increase in the C-fiber response after HFS was blocked in the presence of 2.0 mM AIP (P < 0.05 HFS group versus AIP + HFS group 2 h after conditioning). However, the C-fiber response was not affected by 2.0 mM AIP alone or by vehicle. Thus, our data show that the neuronal process leading to the induction of LTP in the dorsal horn induced by HFS is clearly inhibited by the specific CaMKII inhibitor AIP. It is concluded that CaMKII may be important for the induction of LTP in single nociceptive dorsal horn neurons. PMID- 15804502 TI - Modulation of jaw reflexes induced by noxious stimulation to the muscle in anesthetized rats. AB - Previous studies have shown that jaw reflexes and activity patterns of the jaw muscles were modulated in the presence of jaw muscle pain. However, there is no study comparing the modulatory effects on the jaw reflexes induced by noxious stimulation to the jaw muscle. To clarify this, effects of the application of mustard oil (MO), an inflammatory irritant, into the temporalis (jaw-closing) muscle on (1) jaw-opening reflex evoked by tooth pulp stimulation (TP-evoked JOR) as a nociceptive reflex, (2) jaw-opening reflex evoked by inferior alveolar nerve stimulation as a non-nociceptive reflex and (3) jaw-closing reflex evoked by trigeminal mesencephalic nucleus stimulation as a proprioceptive reflex were investigated in anesthetized rats. The MO application induced suppression of all reflexes, and the effect on the TP-evoked JOR was more prominent than on the other reflexes. To elucidate the involvement of endogenous opioid system for the suppressive effect, a systemic administration of naloxone following the MO application was conducted. The MO-induced suppressive effect on the TP-evoked JOR was reversed by the naloxone administration. The results suggest that noxious stimulation to the jaw muscle modulate jaw reflexes particularly for the nociceptive jaw-opening reflex, and the modulatory effect includes both facilitatory and inhibitory aspects. The results also suggest that pain modulatory systems such as the endogenous opioid system play a crucial role in the suppression of the nociceptive transmissions related to nociceptive reflexes, and in some pathological states, defense reflexes may not be evoked properly. PMID- 15804504 TI - Temporal expression of osteopontin and CD44 in rat brains with experimental cryolesions. AB - Expression of osteopontin and CD44 in the brain was studied after cryolesioning to understand how osteopontin and its receptor, CD44, are involved in processes in the brains of rats with cryolesions. Western blot analysis showed that osteopontin increased significantly at days 4 and 7 post-injury and declined slightly thereafter in cryolesioned brains in comparison with levels in sham operated controls. An immunohistochemical study localized osteopontin in activated microglia/macrophages in the core lesions, where the majority of macrophages proliferate. Osteopontin was also detected temporarily in some neurons and a few astrocytes in the lesion periphery on days 4 and 7 post-injury, but the immunoreactivity in macrophages, neurons, and astrocytes disappeared by day 14 post-injury. There was some CD44, a receptor for osteopontin, in the brain cells of sham-operated rats. After injury, intense CD44 immunostaining was seen in the majority of macrophages and in reactive astrocytes, but not in neurons, in the ipsilateral lesions after day 4 post-injury, and this immunoreactivity remained on day 14 post-injury. These findings suggest that activated microglia/macrophages and some neurons are major sources of osteopontin during the early stage of brain damage induced by a cryolesion and that osteopontin interacts with CD44 expressed on astrocytes and activated microglia/macrophages in the damaged cerebral cortex, possibly mediating cell migration after cryolesioning in the rat brain. PMID- 15804503 TI - Delayed olfactory nerve regeneration in ApoE-deficient mice. AB - Apolipoprotein E (apoE), a lipid transporting protein, is extensively expressed in the primary olfactory pathway, but its function is unknown. We previously reported increased apoE levels in the olfactory bulb (OB) following olfactory epithelium (OE) lesion in mice, and hypothesized that apoE may play a vital role in olfactory nerve (ON) regeneration. To directly test this hypothesis, we examined the rate of ON regeneration following OE lesion in apoE deficient/knockout (KO) and wild-type (WT) mice. OE was lesioned in 2- to 3-month old mice by intranasal irrigation with Triton X-100 (TX). OB were collected at 0, 3, 7, 21, 42, and 56 days post-lesion. OB recovery was measured by both immunoblotting and immunohistochemical analysis of growth cone associated protein (GAP) 43 and olfactory marker protein (OMP). The results revealed that (1) OMP recovery in the OB was significantly slower in apoE KO compared to WT mice; (2) recovery of glomerular area was similarly slower; and (3) GAP43 increases and return to prelesion levels in the OB were slower in KO mice. Together, these results show that olfactory nerve regeneration is significantly slower in KO mice as compared to WT mice, suggesting apoE facilitates olfactory nerve regeneration. PMID- 15804505 TI - Expression of DJ-1 in the adult mouse CNS. AB - Recessively inherited mutations in DJ-1 have recently been linked to familial forms of parkinsonism. However, the regional and cellular expression of DJ-1 is largely unknown. In the present study, we mapped the distribution of DJ-1 transcript and protein by non-radioactive in situ hybridization and immunohistochemistry in adult mouse CNS. For immunohistochemical analysis, we raised a polyclonal antiserum against GST-mouse DJ-1 fusion protein. DJ-1 transcript and protein were strongly and homogenously expressed in all CNS regions. Colocalization experiments revealed expression in neurons of different neurotransmitter phenotypes and in all glial cell types, such as astrocytes, microglia and oligodendrocytes. DJ-1 protein was diffusely distributed throughout the cytoplasma of the soma and the proximal parts of the processes, while the nuclei were always spared. The high expression of DJ-1 in neuronal and glial cells, that is not confined to a single functional system or any anatomical area, supports the view of a basic physiological role in cell biology. PMID- 15804506 TI - Anti-apoptotic effects of muscarinic receptor activation are mediated by Rho kinase. AB - Activation of muscarinic receptors has been shown to be neuroprotective in several different models of apoptosis, but the mechanism of this action is unknown. Therefore, we investigated the intermediate signals mediating the anti apoptotic action of muscarinic receptor activation in SH-SY5Y cells. Inhibition of most muscarinic receptor-coupled actions had no effect on protection, but inhibition of Rho kinase with HA-1077 concentration-dependently was able to completely block the protection against H(2)O(2)- and camptothecin-induced apoptosis produced by stimulation of muscarinic receptors. These results demonstrate that the anti-apoptotic effect provided by muscarinic receptor stimulation is dependent on the activity of Rho kinase. PMID- 15804507 TI - Autoradiographic and immunohistochemical localization of insulin-like growth factor-I receptor binding sites in brain of the brown trout, Salmo trutta. AB - Insulin-like growth factor-I (IGF-I), a peptide closely related to insulin, is known to play crucial roles in brain development. While the central sites of action of IGF-I in higher vertebrates are now well established, surprisingly little is known in the teleost model where the brain undergoes continual, indeterminate, growth. In this study, we have mapped the distribution of putative IGF-I receptor (IGF-IR) binding sites in the brain of the brown trout using both ligand binding in vitro autoradiography and immunohistochemistry. The presence of IGF binding proteins (IGFBPs) was further studied by competitive inhibition using unlabelled IGF-I and des-(1-3)-IGF-I. In both juvenile and adult trout brain, [125I]IGF-I binding was highest in cerebellum and optic tectum, both regions of the teleost brain known to grow the most actively throughout life. At the cellular level, IGF-IR immunoreactivity was confirmed on cell bodies and dendrites, particularly of larger presumptive neurons including purkinje cells and dendritic fibres throughout the molecular layer of the cerebellum. Abundant IGF-IR expression in hypothalamic regions may further be related to neuron growth while a possible hypophysiotropic role will require further investigation. Competitive inhibition studies employing des-(1-3)-IGF-I also suggest IGFBPs are present in all regions exhibiting high [125I]IGF-I ligand binding and confirms the presence of this important regulatory component of the IGF-I system in the teleost brain. The importance of the IGF-I system in brain development, particularly in regions such as the cerebellum, together with the continual lifetime growth of the fish central nervous system, suggest the teleost brain is an extremely useful site for studying the actions of IGF-I in relation to neuron proliferation, growth, and survival in an adult brain. PMID- 15804508 TI - Actions of growth hormone on carbohydrate metabolism and osmoregulation of rainbow trout (Oncorhynchus mykiss). AB - Rainbow trout Oncorhynchus mykiss were injected intraperitoneally with slow release implants of vegetable oil alone or containing ovine growth hormone (oGH) (2 and 5 microgg(-1) body weight), and sampled after 5 days to assess the simultaneous effects of GH on both osmoregulation and carbohydrate metabolism. An enhanced hypoosmoregulatory capacity of oGH-implanted fish is suggested by the increase observed in gill Na+,K+-ATPase activity, and the decrease observed in plasma ion concentration (Na+ and Cl-) and osmolality. GH treatment also elicited increased plasma glucose levels and metabolic changes in liver, gills, kidney, and brain. Major metabolic changes elicited by GH treatment included (1) decreased glycolytic potential and capacity for exporting glucose in liver, (2) enhanced glycogenolytic potential and capacity for use of exogenous glucose in gills and kidney, as well as increased glycolytic capacity in the later tissue, and (3) enhanced glycogenolytic and glycolytic capacities in brain. These metabolic changes elicited by GH treatment support a role for GH in the control of carbohydrate metabolism in salmonids that could be related either to the metabolic changes occurring during osmotic acclimation in nature (a process in which changes in GH levels and carbohydrate metabolism have both been reported) or to metabolic changes associated with growth. PMID- 15804509 TI - Melatonin prevents testosterone-induced suppression of immune parameters and splenocyte proliferation in Indian tropical jungle bush quail, Perdicula asiatica. AB - During reproductive inactive phase (winter months) peripheral high melatonin and low testosterone was noted in male Indian tropical bird, Perdicula asiatica. During this phase exogenous testosterone (1 mg/100 g Bwt./day) suppressed the immune parameters [spleen weight, total leukocyte count (TLC), lymphocyte count (LC), and percent stimulation ratio (% SR)] and depleted splenic cellular density, while melatonin (25 microg/100g Bwt./day) restored the immune parameters and splenic cellular density to the level of control birds. In vitro suppression of splenocyte proliferation by testosterone (3 ng/ml) was also reversed by melatonin (500 pg/ml) supplementation. Therefore, high melatonin in circulation during the reproductive inactive phase acts as immunostimulator and is of high adaptive significance to this bird for survival during the adverse conditions of season and reproductive phase. PMID- 15804510 TI - Pattern of secretion of immunoreactive inhibin/activin subunits by avian granulosa cells. AB - The messenger RNA expression for the inhibin/activin subunits in the granulosa layer of avian follicles of different developmental stages has previously been reported. In the present study, we examined the pattern of secretion of these protein subunits from cultured granulosa cells (GC) of avian follicles of defined maturity. Laying hens were euthanized and the F1, F2, F3, F4, small yellow follicles (SYF; 6-10 mm) and large white follicles (LWF; 3-5 mm) were removed. GC were isolated from the follicles, plated by size at a density of 6.25 x 10(5)cells per well (3 wells per follicle size) and cultured for 48 h in medium 199 with 5% FBS, antibiotics, and 1.0 microg/ml bovine insulin. After 48 h, the cultures were terminated and the media were saved (n = 6 replications). Proteins were precipitated from media, reconstituted for electrophoresis (SDS-PAGE), and analyzed by Western blot. Progesterone was also measured in the medium. For detection of the inhibin alpha-subunit, a rabbit antibody against the chicken inhibin alpha-subunit (1-26 aa) was used. The betaA-subunit was detected with rabbit anti-betaA-subunit (81-113 aa) and the betaB-subunit was detected with rabbit anti-betaB-subunit (80-112 aa). Under reduced conditions, GC from the larger follicle sizes (F1-F4) secreted the most (p < 0.05) immunoreactive inhibin alpha-subunit compared to smaller follicle sizes. Under non-reduced conditions, a band at approximately 32 kDa was detected by both the alpha-subunit antibody and by the betaA-subunit antibody in media from GC of the F1-F4 follicles, suggesting secretion of intact inhibin A. Immunoreactive alpha-subunit and betaB-subunit were detected under reduced conditions in media from the GC of the SYF, suggesting that this follicle population may secrete intact inhibin B. In addition, under non-reduced conditions, cells from the SYF secreted the greatest amount of intact inhibin B (p < 0.05) at a size of approximately 32 kDa. Cells from the LWF expressed low levels of all inhibin subunits. Progesterone concentration in the media from the F1 follicle was greatest and was decreased in media from smaller follicles. It is suggested that the largest follicles in the hierarchy are the source of most circulating intact inhibin A while the small follicles are the source of intact inhibin B. PMID- 15804511 TI - Postnatal ontogeny of erythropoietin and hematology in free-ranging Steller sea lions (Eumetopias jubatus). AB - The hormone erythropoietin (EPO) is responsible for the increased production of red blood cells (RBC) in response to tissue hypoxia. While the role of EPO in hematological development has been established in humans and terrestrial mammals, this relationship has never been examined in marine mammals that rely heavily on stored oxygen to maintain aerobic metabolism while diving. Since blood is the major oxygen storage site in marine mammals, it was hypothesized that EPO may have a significant influence on the development of hematology parameters associated with the expansion of blood oxygen stores during development. To explore this hypothesis, serum EPO concentrations were determined by radioimmunoassay in 235 free-ranging Steller sea lions (Eumetopias jubatus), throughout their Alaskan range. Hematocrit (Hct), hemoglobin (Hb), and red blood cell (RBC) counts were also measured, and mean corpuscular hemoglobin content (MCHC), mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCV) values determined. Erythropoietin and most hematological parameters varied with age. Hematocrit, Hb, RBC, and MCHC decreased after birth, reached their lowest values at two to three months of age, and then increased to values similar to those of adults by five months of age. Since changes in Hct and Hb account for the majority of the changes in blood oxygen stores and EPO was negatively correlated with both, it appears that EPO may play an important role in blood development of Steller sea lions, similar to previous studies on terrestrial mammals. PMID- 15804512 TI - Seasonal expression of LHbeta and FSHbeta in the male newt pituitary gonadotrophs. AB - Seasonal changes in LHbeta and FSHbeta mRNA levels were examined in the pituitary gland of the adult male newt, Cynops pyrrhogaster, using in situ hybridization histochemistry and a quantitative real-time RT-PCR method. The annual fluctuation of LHbeta mRNA and FSHbeta mRNA levels in the pituitary gland displayed a close relationship with seasonal changes in testicular function. The values obtained by both methods showed similar fluctuation. The levels of LHbeta mRNA were always exceeded those of FSHbeta. The present immunoelectron microscopic observations support the data on the gene expression levels of the beta-subunits of LH and FSH. Gonadectomy in the summer increased the LHbeta and FSHbeta mRNA levels. Testosterone replacement inhibited the expression of LHbeta mRNA, but not of FSHbeta mRNA, suggesting that the expression of FSHbeta is regulated by some non steroid factor, probably inhibin. In the case of gonadectomy during any other season, the LHbeta mRNA level increased, but not to the same extent as in summer, and androgen concentrations decreased to the minimum of the year. This finding provides new information about the regulation of annual changes in LHbeta and FSHbeta expression in the pituitary gonadotrophs. PMID- 15804513 TI - A serotonin receptor antagonist, but not melatonin, modulates hormonal responses to capture stress in two populations of garter snakes (Thamnophis sirtalis parietalis and Thamnophis sirtalis concinnus). AB - Hormonal and behavioral responses to a stressor depend on many factors, including the influence of other hormones. We examined the role of melatonin in modulating hormonal responses to capture stress in two populations of male garter snakes, Thamnophis sirtalis. Studies of red-sided (T. sirtalis parietalis) and red spotted (T. sirtalis concinnus) garter snakes were conducted in the field with free-living snakes. Populations of red-sided garter snakes in south-central Manitoba, Canada undergo a period of winter dormancy for approximately 8 months each year followed by an attenuated mating season (4-5 weeks) in early spring. In contrast, the mid-latitude red-spotted garter snake in western Oregon, USA has an extended breeding season and can be active during 10-12 months of the year given appropriate environmental conditions. We chose to study these two populations of garter snakes to investigate possible variation in melatonin function among snakes with different suites of environmental adaptations. To better address these questions, we also examined the effects of 5-hydroxytryptophan (a precursor of melatonin synthesis) and ketanserin (a serotonergic type 2A receptor antagonist) on hormonal responses to capture stress. We observed a trend of increased corticosterone and decreased androgen concentrations in northern latitude red-sided garter snakes (T. sirtalis parietalis) subjected to 4 h of capture stress during the spring. However, these differences were not statistically significant. During the fall, red-sided garter snakes showed no change in corticosterone or androgen concentrations in response to the capture stress treatments. We speculate that northern-latitude red-sided garter snakes suppress hormonal responses to capture stress during preparation for winter dormancy. Treatment with melatonin, 5-hydroxytryptophan, or ketanserin did not significantly influence corticosterone or androgen concentrations of northern latitude red-sided garter snakes during the spring or fall. Mid-latitude red spotted garter snakes (T. sirtalis concinnus) from Oregon showed a statistically significant increase in corticosterone concentrations in response to 4 h of capture stress; treatment with melatonin, 5-hydroxytryptophan, or ketanserin prior to capture stress had no significant influence on plasma corticosterone concentrations. Androgen concentrations of mid-latitude red-spotted garter snakes in response to capture stress were significantly lower than those of non-stressed control snakes. Neither melatonin nor 5-hydroxytryptophan influenced the change in androgen concentrations during capture stress. However, androgen concentrations of snakes treated with ketanserin prior to 4 h of capture stress did not differ significantly from those of non-stressed control snakes. These studies suggest that melatonin does not modulate hormonal responses to capture stress in this ectothermic model. Our results also suggest that a serotonin regulated system may play a role in modulating the activity of the hypothalamic pituitary-gonadal axis during physiological stress responses. PMID- 15804514 TI - Fecal steroid analysis of female giraffe (Giraffa camelopardalis) reproductive condition and the impact of endocrine status on daily time budgets. AB - Gestation and lactation can impose substantial energetic costs on female mammals. We developed a non-invasive means to determine reproductive condition in female giraffe using fecal steroid analysis. Giraffe may be especially challenged during their reproductive cycle because of two characteristics: they are impregnated while lactating and they do not breed seasonally. We studied the social behavior and endocrinology of seven female giraffe in a large naturalistic outdoor enclosure in order to chart connections between maternal physiology and behavior across the reproductive cycle. We found that giraffe gestation averages 448 days among females producing a calf that survived, with fecal pregnane concentrations reaching a zenith during the last trimester of pregnancy. Resumption of ovarian cyclicity following parturition was accelerated after neonatal calf mortality, but ovarian cycles resumed as early as 39 days postparturition while nursing. Although time spent feeding was unaffected by reproductive state, pregnant females significantly reduced time allocated to social behavior and had a tendency to locomote less than when cycling or acyclic. We suggest that modifications in foraging strategies as a function of reproductive state among wild giraffe derive from antipredator activity rather than from metabolic demands. Female giraffe probably cope with simultaneous lactation and gestation by producing high quality milk for neonatal calves commensurate with slow fetal growth and accelerating fetal growth simultaneous with weaning of nursing calves. PMID- 15804515 TI - Molecular characterization of rockfish (Sebastes schlegeli) gonadotropin subunits and their mRNA expression profiles during oogenesis. AB - In an attempt to understand the reproductive regulation in viviparous teleosts, gonadotropin (GTH) subunit cDNAs were characterized and the expression levels of GTH subunit mRNAs in the pituitary glands of the rockfish, Sebastes schlegeli (Scorpaeniformes, Scorpaenidae), were examined by Northern blot analysis. The complete sequences of rockfish GTH subunits (GTHalpha, FSHbeta, and LHbeta) were determined by rapid amplification of cDNA ends (RACE) and nucleotide sequencing. Based on the RACE analysis, the cDNAs of GTHalpha, FSHbeta, and LHbeta consisted of 655, 540, and 529 nucleotides encoding peptides of 132, 130, and 143 amino acids, respectively. The mature peptides of rockfish FSHbeta, LHbeta, and common GTHalpha showed high sequence identities (FSHbeta, 58-62%; LHbeta, 86-94%; and GTHalpha, 87-88%) to those of other perciforme (e.g., orange spotted grouper, red seabream, stripped bass, and sea bass). In a sequence alignment of the mature peptides, rockfish FSHbeta exhibited a unique feature, the lack of a conserved N glycosylation site. This is in contrast to both LHbeta and GTHalpha which contain one and two N-glycosylation sites, respectively, and is consistent with those of other teleosts. The mRNA levels of the GTHalpha subunit increased at the vitellogenic stage and remained steady from ovulation to post-parturition. FSHbeta mRNA levels increased abruptly during the vitellogenic stages and then decreased during ovulation, embryonic development, and post-parturition. LHbeta mRNA levels were observed to rapidly increase during the vitellogenic stage, reached its highest levels during ovulation and was then followed by a decrease at post-parturition. These results suggest that in the female rockfish FSH and LH syntheses are predominant during vitellogenesis and oocyte maturation, respectively. PMID- 15804516 TI - Nucleotide sequence and expression of three subtypes of proopiomelanocortin mRNA in barfin flounder. AB - Melanophore-stimulating hormone (MSH) has been shown to be associated with food intake in addition to body color change in teleosts. MSH is encoded by a proopiomelanocortin (POMC) gene together with endorphin (END). To assess the significance of MSH to biological activities, we determined the structure and evaluated the expression of POMC mRNA in barfin flounder (bf), Verasper moseri, a member of a group of teleosts, Pleuronectiformes. Three subtypes of POMC cDNAs (A, B, and C) were amplified from bf pituitary glands. These bfPOMCs contained segments for N-POMC, alpha-MSH, beta-MSH, and beta-END as do other teleost POMCs, while POMC-C showed remarkable variations in the segments corresponding to N-POMC and beta-END. A phylogenetic tree of ray-finned fish POMCs constructed by the neighbor joining method revealed that the three POMC subtypes may have appeared as a result of duplication events occurring at least twice during the course of bf evolution. The first duplication may have generated the lineage leading to an ancestor of bfPOMC-A and -B and that leading to bfPOMC-C, and then the lineage of bfPOMC-A may have diverged from that of bfPOMC-B. All peptides flanked by processing signals excluding N-POMC-C (1-14) were identified in a single pituitary extract by mass spectrometry, and the cDNAs of three POMCs were amplified from a single pituitary by reverse transcription polymerase chain reaction. These results demonstrated that the three POMC genes are expressed in a single individual. While the bfPOMC-A gene was exclusively expressed in the pituitary, the bfPOMC-B and -C genes were expressed in non-pituitary tissues such as brain, gill, heart, spleen, liver, stomach, intestine, testis, muscle, blood, and skin in addition to the pituitary. The expression levels of the POMC-A, -B, and -C genes in pituitary neurointermediate lobe were greater in the fish reared with a black background than the fish reared with a white background, indicating that MSH derived from all of the three bfPOMC genes was associated with body color change. No difference was observed in the expression levels of bfPOMC-C in the brain in response to feeding status. PMID- 15804517 TI - Recent advances in trichosanthin, a ribosome-inactivating protein with multiple pharmacological properties. AB - Trichosanthin (TCS), a ribosome-inactivating protein extracted from the root tuber of Chinese medicinal herb Trichosanthes kirilowii Maximowicz, has multiple pharmacological properties including abortifacient, anti-tumor and anti-HIV. It is traditionally used to induce abortion but its antigenicity and short plasma half-life have limited the repeated clinical administration. In this review, work to locating antigenic sites and prolonging plasma half-life are discussed. Studies on structure-function relationship and mechanism of cell entry are also covered. Recently, TCS has been found to induce apoptosis, enhance the action of chemokines and inhibit HIV-1 integrase. These findings give new insights on the pharmacological properties of TCS and other members of ribosome-inactivating proteins. PMID- 15804518 TI - The toxicity of microcystin LR in mice following 7 days of inhalation exposure. AB - Microcystins, a family of cyclic heptapeptides produced by the cyanobacteria, Microcystis aeruginosa, have documented hepatotoxic and tumor promoting activities. The purpose of this study was to evaluate the toxicity of inhaled microcystin LR (microcystin). Male BALB/c mice were exposed by nose-only inhalation to 260-265 microg microcystin/m(3) for 7 days. The low-, mid- and high dose groups were exposed for 0.5, 1, and 2h, respectively. Control animals were sham exposed to aerosolized vehicle. Treatment-related microscopic lesions were observed only in the nasal cavity of the mid- and high-dose groups. These lesions consisted of minimal to moderate multifocal degeneration and necrosis of the respiratory epithelium, with variable neutrophilic inflammation and minimal to marked degeneration, necrosis, and atrophy of the olfactory epithelium. The no adverse-effect dose for the nasal lesions was approximately 3 microg/kg body weight, or 20 ng/cm(2) of nasal epithelium. In serum, only two protein peaks, occurring at m/zs of 11,688 and 11,829 Da, exhibited decreases in intensity that were microcystin dose-dependent. While these proteins have not been positively identified, they may be useful in the future as biomarkers of microcystin exposure in humans. PMID- 15804519 TI - Salinity effect on growth and toxin production of four tropical Alexandrium species (Dinophyceae). AB - Four tropical PSP toxins-producing dinoflagellates, Alexandrium minutum, Alexandrium tamiyavanichii, Alexandrium tamarense and Alexandrium peruvianum from Malaysian waters were studied to investigate the influences of salinity on growth and toxin production. Experiments were conducted on constant temperature 25 degrees C, 140 microE mol m(-2) s(-1) and under 14:10 light:dark photo-cycle with salinity ranged from 2 to 30 psu. The PSP-toxin congeners, GTX 1-6, STX, dcSTX, NEO and C1-C2 were analysed by high performance liquid chromatography. Salinity tolerance of the four species in decreasing order is A. minutum>A. peruvianum>A. tamarense>A. tamiyavanichii. Specific growth rates and maximum densities varied among these species with A. minutum recorded as the highest, 0.5 day(-1) and 6 x 10(4) cells L(-1). Toxin content decreased with elevated salinities in A. minutum, the highest toxin content was about 12 fmole cell(-1) at 5 psu. In A. tamiyavanichii, toxin content peaked at optimal growth salinity (20 and 25 psu). Toxin content of A. tamarense, somehow peaked at sub-optimal growth salinity (15 and 30 psu). Results of this study implied that salinity fluctuation not only influenced the growth physiology but also toxin production of these species. PMID- 15804520 TI - Toxic potential of five freshwater Phormidium species (Cyanoprokaryota). AB - Among the Cyanoprokaryota (blue-green algae), the genus Phormidium has thus far rarely been studied with respect to toxin production and potentially resulting human and environmental health effects. We here show that five previously unexplored freshwater species of this genus (Ph. bijugatum, Ph. molle, Ph. papyraceum, Ph. uncinatum, Ph. autumnale) are indeed capable of producing bioactive compounds. Phormidium extracts caused weight loss as well as neuro/hepatotoxic symptoms in mice, and in the case of Ph. bijugatum even death. Very low levels of saxitoxins and microcystins, as confirmed by ELISA, were insufficient to explain this toxicity and the differing toxic potencies of the Phormidium species. Qualitative HPLC analyses confirmed different substance patterns and in the future could aid in the separation of fractions for more detailed substance characterisation. The results in vivo were confirmed in vitro using cells of human, mouse and fish. The fish cells responded least sensitive but proved useful in studying the temperature dependence of the toxicity by the Phormidium samples. Further, the human cells were more sensitive than the mouse cells thus suggesting that the former may be a more appropriate choice for studying the impact of Phormidium to man. Among the human cells, two cancer cell lines were more responsive to one of the samples than a normal cell line, thereby indicating a potential anti-tumour activity. Thus, the five freshwater Phormidium species should be considered in environmental risk assessment but as well, as a source of therapeutic agents. PMID- 15804521 TI - Three structurally related, highly potent, peptides from the venom of Parabuthus transvaalicus possess divergent biological activity. AB - The venom of South African scorpion Parabuthus transvaalicus contains a novel group of peptide toxins. These peptides resemble the long chain neurotoxins (LCN) of 60-70 residues with four disulfide bridges; however they are 58 residues long and have only three disulfide bridges constituting a new family of peptide toxins. Here we report the isolation and characterization of three new members of this mammal specific group of toxins. Dortoxin is a lethal peptide, bestoxin causes writhing in mice and altitoxin is a highly depressant peptide. Binding ability of these peptides to rat brain synaptosomes is tested. While the crude venom of P. transvaalicus enhances the binding of [(3)H] BTX to rat brain synaptosomes none of these individual toxins had a positive effect on binding. Although the primary structures of these toxins are very similar to birtoxin, their 3D models indicate significant differences. Dortoxin, bestoxin and altitoxin cumulatively constitute at least 20% of the peptide contained in the venom of P. transvaalicus and contribute very significantly to the toxicity of the venom of this medically important scorpion species. Therefore the amino acid sequences presented here can be used to make more specific and effective antivenins. Possible approaches to a systematic nomenclature of toxins are suggested. PMID- 15804522 TI - Identification of a novel pulmonary oedema producing toxin from Indian red scorpion (Mesobuthus tamulus) venom. AB - The experiments were conducted to identify the toxin that produces pulmonary oedema in Mesobuthus tamulus (BT) envenomed animals. Crude BT venom was subjected to Sephadex gel filtration (G-75) and the fractions were screened for optical density (OD), neurotoxicity (prolongation of compound action potential in frog sciatic nerve) and lethality. All these parameters exhibited a peak between 54-94 ml eluates. Fractions of this peak were pooled (SP) and loaded on to carboxymethyl cellulose column. The column was then eluted with increasing buffer concentrations at constant pH and temperature. Eluates were screened for neurotoxicity and OD. Four peaks of neurotoxic activity (T1-T4) were detected. T2 and T3 were lethal whereas T1 and T4 were non-lethal. T2 exhibited mainly neurotoxicity and failed to augment phenyldiguanide (PDG)-induced reflex response or to produce pulmonary oedema. T3 was having minimal neurotoxic actions but augmented PDG-reflex and produced pulmonary oedema. The effects of T3 persisted even after dialysis with 8 kDa cut-off filter but not those of T2. The T3 effects resembled toxic manifestations of BT venom and were blocked by aprotinin pre treatment. T3 demonstrated a band at approximately 100 kDa in SDS-PAGE. The results demonstrate the presence of a lethal, high molecular weight, pulmonary oedema producing toxin in BT venom. PMID- 15804523 TI - Degradation of microcystin-RR by UV radiation in the presence of hydrogen peroxide. AB - Experiments were conducted to investigate the degradation of microcystin-RR in order to assess the effectiveness and feasibility of the combined UV/H(2)O(2) catalytic system for purification of water polluted by microcystins. The operating parameters such as hydrogen peroxide dosage, pH value, UV light intensity, initial concentration of microcystin-RR and reaction time were evaluated, respectively. The degradation efficiency increased nonlinearly with increasing UV light intensity and hydrogen peroxide dosage, respectively. There existed an optimal hydrogen peroxide dosage, beyond which the reagent exhibited an inhibitory effect, for degrading microcystin-RR. The degradation process could be fitted by both of the pseudo-first-order and second-order kinetics well and primarily followed a mechanism of both direct photolysis and hydroxyl radical oxidation. Compared with the treatment using UV radiation and hydrogen peroxide individually, the combined UV/H(2)O(2) system could significantly enhance the degradation efficiency due to the synergetic effect between UV radiation and hydrogen peroxide oxidation. The observed rate constants decreased and the corresponding half-lives prolonged as the concentrations of microcystin-RR increased. The combined UV/H(2)O(2) process provides an effective technology for the removal of microcystins from drinking water supplies. PMID- 15804524 TI - Insecticidal effects of canatoxin on the cotton stainer bug Dysdercus peruvianus (Hemiptera: Pyrrhocoridae). AB - Canatoxin (CNTX) is a variant form of urease isolated from Canavalia ensiformis (Leguminosaea) seeds. A possible role in the plant defense was proposed for CNTX, due to its toxicity upon feeding to the beetle Callosobruchus maculatus, and the hematophagous bug, Rhodnius prolixus. The toxic effect is caused by a canatoxin derived peptide ( approximately 10kDa) formed by insect cathepsin-like digestive enzymes. In order to evaluate their potential as bioinsecticides, the effects of CNTX and its peptide were evaluated on a phytophagous hemipteran insect Dysdercus peruvianus, a pest of cotton culture. For the bioassays, the insects fed on gelatin capsules containing powdered cotton seeds, mixed with the freeze-dried protein and other test materials and were observed for survival rate, weight gain and molting. Ingestion of canatoxin, or a recombinant 10kDa peptide derived from it, severely affected young forms of the insects, delaying their development or leading to their death. In contrast, adults were insensitive to diets containing higher concentrations of canatoxin. Cathepsin-like proteinases predominated and showed distinct pattern of enzymatic activities in midguts homogenates according to the developmental stage of the insect, a fact which may explain the different susceptibility of nymphs as compared to adult D. peruvianus. The data presented confirm the potential use of canatoxin-like proteins and derived peptides as bioinsecticides. PMID- 15804525 TI - Biochemical and physiological analyses of a hemolytic toxin isolated from a sea anemone Actineria villosa. AB - A species of venomous sea anemone Actineria villosa was recently found inhabiting the coastal areas of Okinawa, Japan. This marine animal produces various proteinous toxins, so that a local health organization was called for medical treatment for those who had accidental contact with this animal. In this study we analyzed the biochemical and physiological properties of hemolytic protein from A. villosa. The toxin purified from the tentacles of the animals was found to be a protein with a molecular weight of approximately 19 kDa. We named this newly found hemolytic toxin of A. villosa, Avt-I. Incubation of the toxin with sphingomyelin inhibited hemolytic activity by up to 85%, showing that Avt-I may target sphingomyelin on the erythrocyte membrane. The hemolytic activity was stably maintained at temperatures below 45 degrees C, however, a sharp linear decrease in heat stability was observed within the range of 45-55 degrees C. Our results provide the first evidence that A. villosa produces a toxin with strong hemolytic activity similar in biochemical and physiological properties to other members of actinoporin family previously isolated from related species of sea anemones. PMID- 15804526 TI - Induction of apoptosis/necrosis in various human cell lineages by Haemophilus ducreyi cytolethal distending toxin. AB - We investigated the impact of highly purified Haemophilus ducreyi cytolethal distending toxin (HdCDT) on the apoptosis and necrosis of various human cells; including myeloid cells, epithelial cells, keratinocytes, and primary fibroblasts. The levels of apoptosis and necrosis induced in these cells were compared to those induced by HdCDT in human T cells and in the Jurkat T cell line. Levels of caspase-3 activity were measured, and membrane changes like phosphatidylserine (PS) translocation was evaluated after double-staining with the fluorescein isothiocyanate (FITC)-labeled annexin V and propidium iodide (PI) using flow cytometry. HdCDT induced various degrees of apoptosis and necrosis in dose- and time-dependent manners in cells of various lineages. Early and late apoptosis (annexin V-stained cells) were induced in more than 90% of T cells and monocytes after treatment with 100 ng/ml HdCDT for 24 and 48 h, respectively. The corresponding numbers for epithelial cells, keratinocytes, and fibroblasts were 26-32% after treatment with 100 ng/ml HdCDT for 48 h. HdCDT appears to eliminate effectively by inducing apoptosis those cells that are involved in immune responses. Epithelial cells, keratinocytes and fibroblasts, which are important for the healing of chancroid ulcers, are eliminated by apoptosis or necrosis after contact with HdCDT, albeit slower and to a lesser extent than T cells. PMID- 15804527 TI - Antimicrobial activity of the bufadienolides marinobufagin and telocinobufagin isolated as major components from skin secretion of the toad Bufo rubescens. AB - The increase in the emergence of antibiotic-resistant microorganisms and difficult to treat infections caused by these pathogens stimulate research aiming the identification of novel antimicrobials. Skin secretion of amphibian contains a large number of biologically active compounds, including compounds that performance defense mechanisms against microorganisms. In the present work, two antimicrobial bufadienolides, telocinobufagin (402.1609 Da) and marinobufagin (400.1515 Da), were isolated from skin secretions of the Brazilian toad Bufo rubescens. The specimens were collected in Brasilia (Distrito Federal, Brazil), the skin secretions extracted by electric stimulation, and submitted to purification by RP-HPLC. The molecular structure and mass determination were done by (1)H and (13)C NMR and mass spectrometry data, respectively. The antimicrobial activity was performed by liquid growth inhibition against Staphylococcus aureus and Escherichia coli. The minimum inhibitory concentrations of telocinobufagin and marinobufagin were, respectively, 64.0 and 16.0 microg/mL for E. coli and both 128 microg/mL for S. aureus. Besides the antimicrobial activity both bufadienolides promoted an increase of the contraction force in isolated frog ventricle strips. PMID- 15804528 TI - Selective detection of saxitoxin over tetrodotoxin using acridinylmethyl crown ether chemosensor. AB - At pH 7.1, saxitoxin decomposes to produce a trace impurity that can interfere with fluorescence sensing when using irradiation wavelengths near 325 nm. The fluorophore acridine is found to be a suitable component of arylmethyl crown ether chemosensors for the fluorescent detection of saxitoxin. These sensors are selective for the detection of saxitoxin over tetrodotoxin. PMID- 15804529 TI - Functional, cellular and molecular aspects of skeletal muscle recovery after injury induced by snake venom from Notechis scutatus scutatus. AB - We have analysed the rate and ultimate extent of muscle functional recovery after snake venom-induced myotoxicity, as well as the relationships between functional, biochemical and structural indices of recovery. We also compared the effects of various injuries leading to muscle necrosis, loss of innervation/vasculature and/or precursors of muscle cells (pmc). We found that several parameters of rat soleus muscle such as maximal isometric force, slow myosin heavy chain, and citrate synthase, were fully and rapidly restored within 6 weeks after treatment with snake Notechis scutatus venom (im, 2 microg/muscle). In contrast, some muscle contractile properties (degree of tetanic fusion, fatigue resistance...) were not fully recovered even by 12 weeks after venom treatment. However, when compared to other injuries, recovery 3 weeks after venom treatment, was better than that observed after severing the terminal nerve and accompanying vessels and after cryodamage known to kill pmc. In conclusion, our studies demonstrate that contrary to what is commonly believed -- muscle treated by myotoxic agent does not recover rapidly and fully. However, the degree or rate of muscle recovery after snake venom treatment was much better when compared to other types of injury. In addition, histological and biochemical parameters cannot be used as such to easily predict functional recovery following injury. PMID- 15804530 TI - Specific identification of Lachesis muta muta snake venom using antibodies against the plasminogen activator enzyme, LV-PA. AB - Sandwich-type enzyme linked immunosorbent assays (ELISA) were developed to detect Lachesis muta muta (bushmaster) snake venom using antibodies against the plasminogen activator enzyme (LV-PA). Antibodies to LV-PA were obtained by immunization of one rabbit with the purified enzyme. The IgG fraction was purified from rabbit blood in a single step on a column of Sepharose-L. m. muta venom and used to coat the microtiter plates. The specificity of the assay was demonstrated by its capacity to correctly discriminate between the circulating antigens in mice that were experimentally inoculated with L. m. muta venom from those in mice inoculated with venoms from Bothrops atrox, B. brazili, B. castelnaudi, Bothriopsis taeniata, B. bilineata, Crotalus durissus ruruima and the antigenic Bothrops (AgB) and Crotalus (AgC) pools venoms used to produce Bothropic and Crotalic antivenoms at Fundacao Ezequiel Dias (FUNED). Measurable absorbance signals were obtained with 1.5 ng of venom per assay. The ELISA was used to follow the kinetic distribution of antigens in experimentally envenomed mice. PMID- 15804531 TI - Molecular basis of the low activity of antitumor anthracenediones, mitoxantrone and ametantrone, in oxygen radical generation catalyzed by NADH dehydrogenase. Enzymatic and molecular modelling studies. AB - Synthetic antitumor anthracenedione drugs, in contrast to anthracycline antibiotics, are ineffective in free radical formation in NADH dehydrogenase system. Our results have indicated that neither the reduction potential nor the side chain conformation and the energies of border orbitals (HOMO and LUMO) determine the ability of anthracenediones to stimulate reactive oxygen species formation in NADH dehydrogenase system. It was shown that the distribution of the molecular electrostatic potential (MEP), around the quinone system was crucial for this ability. We have found for non-stimulating anthracenediones that the clouds of positive MEP cover the quinone carbon atoms while for agents effective in stimulating reactive oxygen species formation the clouds of negative MEP cover continuously the aromatic core together with the quinone system. PMID- 15804532 TI - A structure-activity relationship study of quinone compounds with trypanocidal activity. AB - A set of 25 quinone compounds with anti-trypanocidal activity was studied by using the density functional theory (DFT) method in order to calculate atomic and molecular properties to be correlated with the biological activity. The chemometric methods principal component analysis (PCA), hierarchical cluster analysis (HCA), stepwise discriminant analysis (SDA), Kth nearest neighbor (KNN) and soft independent modeling of class analogy (SIMCA) were used to obtain possible relationships between the calculated descriptors and the biological activity studied and to predict the anti-trypanocidal activity of new quinone compounds from a prediction set. Four descriptors were responsible for the separation between the active and inactive compounds: T5 (torsion angle), QTS1 (sum of absolute values of the atomic charges), VOLS2 (volume of the substituent at region B) and HOMO-1 (energy of the molecular orbital below HOMO). These descriptors give information on the kind of interaction that occurs between the compounds and the biological receptor. The prediction study was done with a set of three new compounds by using the PCA, HCA, SDA, KNN and SIMCA methods and two of them were predicted as active against the Trypanosoma cruzi. PMID- 15804533 TI - Carbapenem-based prodrugs. Design, synthesis, and biological evaluation of carbapenems. AB - Syntheses of racemic trans-3-hydroxycarbonyl-6-(phenylacetamido)carbapenem (13), trans-3-phosphono-6-(phenylacetamido)carbapenem (17), and beta-lactam based prodrugs 19 and 22 were accomplished. Carbapenem 13 was found to possess antibacterial activity, comparable with imipenem (+)-3, against Staphylococcus aureus FDA 209P, S. aureus 95, Escherichia coli ATCC 39188, Klebsiella pneumoniae NCTC 418, Pseudomonas aeruginosa 1101-75, P. aeruginosa 18S-H, and Xanthomonas maltophilia GN 12873. Like imipenem ((+)-3), carbapenem 13 was not stable to X. maltophilia oxyiminocephalosporinase type II. Its phosphonate analog 17, however, was neither a significant antibacterial agent nor a good beta-lactamase inhibitor. Chemical combinations of trans carbapenem 13 with cis carbapenem 6 (compound 19) as well as clavulanic acid (20) with cis carbapenem 6 (compound 22) via a tetrachloroethane linker exhibited remarkable activity against beta lactamase producing microorganisms in vitro. PMID- 15804534 TI - Synthesis and antimycobacterial activities of glycosylated amino alcohols and amines. AB - Reduction of glycosyl beta-amino esters (6-14 and 25-30) with lithium aluminum hydride resulted in glycosyl amino alcohols (15-23 and 31-36) in good yields. However, reductive amination of glycosyl aldehydes (1-3) with different amines in presence of sodium borohydride resulted in good to moderate yields of glycosyl amines (37-41). All the compounds were evaluated for antitubercular activity against Mycobacterium tuberculosis H37Ra and H37Rv. Compounds 18, 21, 35 and 36 exhibited antitubercular activities with MIC ranging from 6.25 to 3.12 microg ml( 1). PMID- 15804535 TI - Benzenesulfonamide analogs of fluoroquinolones. Antibacterial activity and QSAR studies. AB - The structure-activity relationships (SAR) of new antibacterial benzenesulfonamidefluoroquinolones (BSFQs), coming from derivatization of N4 piperazinyl of ciprofloxacin (CIP) were studied. The behavior of the new BSFQ series was similar to the previously norfloxacin (NOR) analogs reported, making possible a quantitative structure-activity relationships (QSAR) analysis of the complete set of BSFQs. The presence of the benzenesulfonylamido (BS) groups shifted the activity of classic antimicrobial fluoroquinolones from being more active against Gram-negative to Gram-positive strains. QSAR studies through Hansch analysis showed a linear correlation of the activity with electronic and steric parameters. Small electron-donor groups would increase the in vitro activity against Gram-positive bacteria. Hydrophobic properties played a minor role when activity is measured as minimum inhibitory concentration (MIC). QSAR analysis also reinforces previous biological findings about the presence of new interactions with target topoisomerases. PMID- 15804536 TI - Synthesis, pharmacological activity and hydrolytic behavior of glyceride prodrugs of ibuprofen. AB - For reducing the gastrointestinal toxicity associated with ibuprofen its carboxylic group was condensed with the hydroxyl group of 1,2,3-trihydroxy propane 1,3-dipalmitate/stearate to give the ester prodrugs 3a and 3b. The release of ibuprofen from these prodrugs has been studied at pH 3, 4, 5 and 7.4 by HPLC using methanol and 0.05% phosphoric acid (80%) (70:30) as mobile phase. The prodrugs showed insignificant hydrolysis at pH 5 compared to pH 7.4 indicating that the prodrugs do not break in stomach but release ibuprofen at pH 7.4 in adequate amounts. In vivo hydrolysis studies in rats, the peak plasma concentration of ibuprofen was attained in 1.5 h in case of ibuprofen and in 2 h in prodrugs treated animals. The plasma concentration was found to be less at all times in animals treated with ibuprofen compared to the prodrugs treated animals. The maximum anti-inflammatory activity of ibuprofen was observed at 2 h whereas prodrugs showed maximum activity at 3 h and remained practically constant upto 8 h whereas a decrease in activity was observed with free ibuprofen. Further the prodrugs showed less gastric ulcers compared to ibuprofen. An average score of 0.16, 0.45, 0.97 and 0.20, 0.76, 1.02 of ulcers was observed with 3a and 3b compared to an average score of 0.75, 1.10, and 2.97 with ibuprofen. These prodrugs also showed significant protection against acetic acid induced writhings in rats. These finding suggested that both the prodrugs are better in action as compared to the parent drug and are advantageous in having less gastrointestinal side effects. PMID- 15804537 TI - Synthesis, molecular structure, and in vitro antitumor activity of new 4-chloro-2 mercaptobenzenesulfonamide derivatives. AB - The reaction of 3-amino-2-(2-alkylthio-4-chlorobenzenesulfonyl)guanidines 2a-j with 1,2-dicarbonyl compounds are described. Depending on structure of 1,2 dicarbonyl reagent novel 2-alkylthio-5-chloro-N-(1,2,4-triazin-3 yl)benzenesulfonamides 3-15, 1-(2-alkylthio-4-chlorobenzenesulfonyl)-3-(2 oxobutane-3-ylidenoimino)guanidines 16-18 and 2-alkylthio-4-chloro-N-(1,2 dihydroxycyclobuta[e]1,2,4-triazin-3-yl)benzenesulfonamides 19-21 are obtained. The structures of these compounds were confirmed on the basis of elemental analysis, spectral data and X-ray analysis. The compounds 4, 5, 7, 9, 10, 12-15, 17, 18 and 20 were screened at the National Cancer Institute (NCI) for their in vitro activities against a panel of 56 tumor cell lines and relationship between structure and antitumor activity are discussed. The compounds 10, 12, 17 and 20 were inactive, whereas the other compounds exhibited reasonable activity against one or more human tumor cell lines. The prominent compound 18 showed significant activity against cell lines of colon cancer (HCT-116), renal cancer (786-0) and melanoma (M14) (GI50 in the range 0.33-1.08 microM) as well as good selectivity toward non-small cell lung cancer (HOP-62) cells (GI50 = 0.05 microM, TGI = 0.38 microM and LC50 = 4.83 microM). PMID- 15804538 TI - Novel pyrrolinones as N-methyl-D-aspartate receptor antagonists. AB - A series of oximes, deriving from 2-arylidene-pyrroline-3,4-diones (7, 8, 22, 23) has been prepared. The presence of tautomers in their solutions has been established by spectroscopic means. The compounds reacted with diazomethane chiefly by N-methylation forming nitrones (10, 11). The analogously prepared 2 arylidene-4-nitropyrrolin-3-ones (12, 13, 24, 25), formally derived from nitrotetramic acids, yielded nitronic acid esters (14, 15, 26) upon reaction with diazomethane. The structures were elucidated by spectral evidence and-in the case of compounds 10 and 20b-by X-ray diffraction analysis. The binding affinity of some of the new compounds toward the N-methyl-d-aspartate (NMDA) (glycine site) receptor has been measured thus providing the basis for further structure activity relationship studies. Oxime 8b showed the highest binding potency (Ki= 9.2 microM). PMID- 15804539 TI - Antibacterial and antifungal activities of new pyrazolo[3,4-d]pyridazin derivatives. AB - Several new pyrazolo[3,4-d]pyridazin derivatives were prepared by the reaction of two [corrected] 1H-pyrazole-3-carboxylic acids and various hydrazines. The compounds were tested for antimicrobial activities against Gram-negative, Gram positive bacteria and fungi. The compounds named as 7e, f had the highest antimicrobial activities against Gram-negative, Gram-positive bacteria and fungi with minimum inhibitory concentrations in the range of 0.31 to<0.0024 mg ml(-1) . PMID- 15804540 TI - Synthetic studies on cyclic octapeptides: Yunnanin F and Hymenistatin. AB - Two biologically active cyclic peptides, Yunnanin F 8 and Hymenistatin 16 were synthesized and the structures were established on the basis of analytical, IR, NMR and mass spectral data. The newly synthesized compounds were screened for their antimicrobial and pharmacological activities. These cyclic octapeptides have shown moderate to good growth inhibition against bacterial strains and weak activity against fungal strains more than that of the standard drug against only Pseudomonas aeruginosa but weak to moderate activity against remaining three bacterial strains. They have shown very weak activity against fungal strains. Yunnanin F possessed good anthelmintic activity while Hymenistatin possessed very low activity, but both showed moderate anti-inflammatory activity. PMID- 15804542 TI - Prepulse inhibition of perceived stimulus intensity: paradigm assessment. AB - In 1939, Helen Peak reported that the presentation of paired acoustic stimuli, separated by 177ms, resulted in a 25% reduction in the perceived intensity of the second stimulus. After 65 years, this form of prepulse inhibition of perceived stimulus intensity (PPIPSI) remains relatively less developed, compared to technically and analytically complex "gating" measures, e.g. PPI of startle and P50 event related potential (ERP) suppression. We report that unlike PPI, which can be observed at very brief inter-stimulus intervals (ISI's), PPIPSI requires ISI's >or=60ms. It exhibits significant test-retest stability over a 2-week period, does not exhibit sex differences or menstrual cyclicity, and is not significantly associated with personality dimensions of novelty seeking, harm avoidance, reward dependence or sensation seeking. PPIPSI is a robust, stable, parametrically sensitive and "low tech" measure of sensory gating that is relatively unaffected by a number of biological variables in normal individuals. PMID- 15804541 TI - Synthesis of novel amino and acetyl amino-4-methylcoumarins and evaluation of their antioxidant activity. AB - The six novel 4-methylcoumarins bearing different functionalities such as amino, hydroxy, N-acetyl, acetoxy and nitro have been synthesized and confirmed on the basis of their spectral data (1H-, 13C-NMR, UV, IR and EI mass). They were examined for the first time for their effect on NADPH dependent liver microsomal lipid peroxidation in vitro, and the results were compared with other model 4 methylcoumarin derivatives to establish the structure-activity relationship. Our studies demonstrated that amino group is an effective substitute for the hydroxyl group for antioxidant property and produced a dramatic inhibition of lipid peroxidation. Ortho dihydroxy and ortho hydroxy-amino coumarins were found to possess highest antioxidant and radical scavenging activities. PMID- 15804543 TI - Socioeconomic position and cardiovascular and neuroendocrine responses following cognitive challenge in old age. AB - Social disparities in health persist into old age, and differences in psychophysiological responsivity may contribute to this pattern. We assessed whether higher socioeconomic status (SES) is associated with attenuated cardiovascular and neuroendocrine responses elicited by cognitive tasks in old age. We tested 132 community-dwelling men and women aged 65-80 years, divided on the basis of educational attainment into higher and lower SES groups, and compared them with 26 higher educated participants aged 27-42 years. Blood pressure, hemodynamic variables and salivary cortisol were assessed in response to the performance of three cognitive tasks, and then during recovery. Older groups showed smaller heart rate and larger cortisol changes than younger participants. Post-task recovery in heart rate, stroke volume, pre-ejection period, and systolic blood pressure was greatest in the younger group, least in the older/lower education group, and intermediate in the older/higher education group. SES did not influence the increased cortisol responsivity of older participants. The results are consistent with the notion that higher SES protects against age-related changes in cardiovascular response profiles, particularly during recovery. PMID- 15804544 TI - Heightened cortisol responses to daily stress in working women at familial risk for breast cancer. AB - Consistent with animal models and experimental studies with humans facing other 'background' stressors, women at familial risk for breast cancer have been reported to have stronger cortisol responses to laboratory stressors. To explore the relevance of these findings to daily life, we compared work-stress cortisol responses in women with >or=1 first-degree relative with breast cancer (FH+, n = 74) to women without this risk factor (FH-, n = 141). Repeated-measures ANOVA revealed a group by time interaction (p 30 minutes after stress termination) relative high levels of cortisol inhibit approach motivation. PMID- 15804546 TI - Stroop color-word test, arousal, electrodermal activity and performance in a critical driving situation. AB - The aim of this experiment was to study drivers' performance when confronted with a critical crash avoidance situation. Subjects were asked to cover three laps of a private circuit (4 min per lap), respecting speed limits. During the last lap, an inflated dummy car, placed at an intersection, was pulled onto the traffic lane. The synchronization signal releasing the obstacle was triggered to make the braking distance too short, thus requiring subjects to brake and turn the driving wheel simultaneously. Before driving, subjects completed the Stroop color-word test. Skin conductance (SC) was recorded continuously, before and during driving. Subjects who performed the least well also performed the least well to Stroop test. The SC level showed that subjects avoiding the obstacle were more aroused than those who crashed into the dummy car. Performance to Stroop test and physiological arousal were thus shown to be determining factors in management of a critical driving situation. PMID- 15804547 TI - Odor mixture alters neural resources during symbolic problem solving. AB - The smell of a cup of coffee is produced by many different odor chemicals combined in a mixture, yet the perception of that odor is of a single unified whole. Recent evidence has demonstrated that mixtures of odors share some of the same spatiotemporal features of speech sounds and may use similar brain resources in associating those features with the symbols they represent. This experiment investigated the hypothesis that an odor mixture would interfere with a math task that requires symbolic but not spatial processing. Results indicated the pattern of brain electrical activity was similar for the single odors and the mixture during spatial processing. During solution of the task requiring symbolic processing, the odor mixture produced a pattern of brain electrical activity different from the single odorants. These data suggest that the perception of odor mixtures may use some of the same resources associated with symbolic processing. PMID- 15804548 TI - Causal coherence analysis of heart rate variability and systolic blood pressure variability under mental arithmetic task load. AB - Causal coherence analysis based on a closed-loop bivariate autoregressive model was applied to heart rate variability and systolic blood pressure (SBP) variability during mental arithmetic tasks to clarify how mental task load affects the linear closed loop interaction between cardiac and vascular systems. Thirteen normal male subjects performed a mental arithmetic task, button press task, and rest task while measuring their RR interval (RRI) and SBP. The mean value in the low frequency (LF) band (0.04-0.15Hz) of the squared causal coherence function from SBP to RRI during the mental arithmetic task was significantly higher than during the other two control tasks. Conversely, the LF band of the squared causal coherence function from RRI to SBP during the mental arithmetic task tended to be lower than during the rest task. These results suggest that mental arithmetic tasks enhance linear causal coupling from the vascular to cardiac system, and conversely weaken that from the cardiac to vascular system. PMID- 15804549 TI - Age-related effects of novel visual stimuli in a letter-matching task: an event related potential study. AB - Younger and older participants (n=8 in each age group) performed a letter matching task, where they had to respond to identical letters (P=0.15). Task relevant (target and non-target) letters were presented on two corners of an imagery square, while on the other two corners irrelevant letters were presented. In some trials (P=0.05), pictures of visual objects (novels, unrelated to the matching task) were presented. Reaction times were slower and error rates higher for older adults. The amplitude of the N1 event-related potential component was smaller over anterior locations for the older adults. All target stimuli elicited a late positivity over anterior locations for both age groups, but the late positivity (P3b) was absent over posterior scalp for the older adults. For the younger adults, novel stimuli elicited a large negative component (N2(novel)) that was maximal over the central location. No such activity was observed for older adults. Based on the results, processing of novel stimuli is considered to be compromised in the elderly. PMID- 15804551 TI - Elder abuse and neglect: an overview. AB - The dramatic growth of the American elderly population has great implications for our health care system. The "demographic imperative" that has fueled the awareness of the needs of older adults has a major impact on issues related to social welfare, justice, and economics. There are 45 million people over the age of 60 and 3 million over the age of 85. Those over age 85 represent the fastest growing segment of the elderly population. With this trend comes a segment of the population that is at risk for abuse, neglect, or self-neglect. We are challenged to be aware of the many faces of elder mistreatment and to understand it in the broader context of domestic violence. All health care professionals working with older adults need to become familiar with the recognition, treatment, and prevention of elder abuse and neglect. PMID- 15804552 TI - Medical implications of elder abuse and neglect. AB - Recognition of elder abuse and neglect among health care professionals has been a relatively recent phenomenon. Each year, millions of elderly persons suffer as the result of abuse and neglect. Their quality of life is severely jeopardized in the form of worsened functional status and progressive dependency, poorly rated self-health, feelings of helplessness, and from the vicious cycle of social isolation, stress and further psychologic decline. Other medical implications of abuse and neglect include higher health systems use in the form of frequent ER visits, higher hospitalization, and higher nursing home placement; most importantly, it is an independent predictor for higher mortality. Physicians are well situated in detecting and reporting suspected cases and taking care of the frail elders who are victims of abuse and neglect, but there are barriers on the individual level, and there is a broader need for system change. Through education, training, and reinforcement, there are strategies to get health care professionals more involved and provide effective management protocols and guidelines for us to advocate for our patients in the current epidemic of elder abuse and neglect. PMID- 15804553 TI - Dementia and elder abuse. AB - Dementia and elder abuse are relatively common and under-diagnosed geriatric syndromes. A unique relationship is observed when the two entities coexist. Special issues can confound the care of the dementia patient suspected of being abused. Impaired language or motor abilities to communicate abusive situations to a third party, lack of decisional capacity to address the abusive situation, disinhibited behavior that contributes to a cycle of violence, and coincident depression of the abused elder complicate the diagnosis and management of elder abuse. Education of the caregiver and attention to caregiver stress, including depression, may prevent onset and perpetuation of abuse. PMID- 15804554 TI - Elder abuse and neglect in long-term care. AB - Patient and caregiver education and other interventions targeted toward risk factors or types of abuse or neglect play an invaluable role in preventing elder abuse and mistreatment. PMID- 15804555 TI - Cultural issues and elder mistreatment. AB - The article is a brief discussion of a few of the complex issues that may represent a cultural component of elder mistreatment. Reference is made to historical perspectives and subcultures. The goals of the article are to broaden the discussion around the issues of elder abuse in various population groups and to consider some methods of intervention. PMID- 15804556 TI - When elders lose their cents: financial abuse of the elderly. AB - Elder financial abuse is one of the most difficult types of elder abuse to diagnose given its lack of overt physical symptomatology. Unusual bank account activity, sudden changes of beneficiaries or agents in estate planning or advance directive documents, and worsening of medical conditions due to lack of follow-up or unfilled prescriptions are indicators of potential financial abuse. Federal laws are aimed at education and prevention (not treatment), whereas state laws are more comprehensive. Criminal and civil legal remedies exist to bring the abuser to justice, but the emotional trauma of financial exploitation lasts a long time. PMID- 15804557 TI - The legal and governmental response to domestic elder abuse. AB - Older Americans constitute the fastest growing segment of the United States population and may account for 20% of the Unites States population by 2050. The federal government has taken minimal action to identify and solve their problems. Due to the federal government's inaction, states have become the primary engine for combating abuse. This is most often seen through adult protective services, which primarily consist of mandatory reporting laws, involuntary interventions, and educational programs. Funding is the primary roadblock to the successful execution of state laws targeting domestic elder abuse. The proposed federal Elder Justice Act of 2003, if passed, may fill in the gaps of current federal legislation by implementing a uniform method of response to domestic elder abuse and providing funding to the states to rectify instances of abuse. PMID- 15804558 TI - Forensic markers in elder female sexual abuse cases. AB - Evidence exists that older adults are victims of sexual assault and rape. The scope of the problem, prevalence, and correlates of these sex crimes are relatively unknown. Such knowledge deficits are major barriers to detecting, prosecuting, and preventing sex crimes against older adults. Understanding how intentional sexual injuries are inflicted on older adults is a growing concern as the population over 65 increases. This study contributes critical information to guide the identification of physical and psychologic markers of elder sexual abuse to be integrated by clinicians and law enforcement as forensic medical evidence. PMID- 15804559 TI - Core data elements tracking elder sexual abuse. AB - Sexual abuse in the older adult population is an understudied vector of violent crimes with significant physical and psychological consequences for victims and families. Research requires a theoretical framework that delineates core elements using a standardized instrument. To develop a conceptual framework and identify core data elements specific to the older adult population, clinical, administrative, and criminal experts were consulted using a nominal group method to revise an existing sexual assault instrument. The revised instrument could be used to establish a national database of elder sexual abuse. The database could become a standard reference to guide the detection, assessment, and prosecution of elder sexual abuse crimes as well as build a base from which policy makers could plan and evaluate interventions that targeted risk factors. PMID- 15804560 TI - Community approaches to elder abuse. AB - Collaboration of professionals from diverse disciplines is required to address the multiple, complex issues and needs of community-dwelling elders who are abused or neglected. Reporting suspected elder abuse or neglect cases to Adult Protective Service (APS) agencies provides access to services that address the social, medical, and legal needs of elderly persons. A geriatric interdisciplinary team can provide a comprehensive medical, functional, and social assessment. Based on the findings from the assessment and in collaboration with the APS team, the intervention plan can be formulated. Some cases of elder abuse or neglect may require intervention from the criminal justice or the civil justice system for serious legal issues such as sexual assault, financial exploitation, or guardianship. Other resources, such as Area Agencies on Aging, local women's shelters, and The National Center for Elder Abuse, are available to help manage elder abuse and neglect cases in the community. PMID- 15804561 TI - Elder abuse, neglect, and exploitation: policy issues. AB - Elder abuse remains a rapidly growing but largely invisible national policy issue. As the number of elderly persons increases, so will elder abuse, neglect, and financial exploitation. This has implications not only for the victims and the programs struggling to protect them but also for publicly funded programs such as Medicare and Medicaid. The urgent problem is to address elder abuse on a national level in a comprehensive and informed way to prevent the untold suffering of hundreds of thousands of older persons who deserve to live their final years with dignity and security. PMID- 15804562 TI - Ligand-binding and signaling properties of the Ax[M1] form of Notch. AB - The Abruptex class of Notch alleles has attracted interest because they exhibit some properties that are best explained in terms of increased activity and others that are best explained in terms of reduced activity in vivo. Here, we report a comparison of the properties of Abruptex[M1] and wild-type Notch as ligand binding receptors. Abruptex[M1] showed less activity than wild-type Notch in its ability to bind Delta and Serrate and was expressed at reduced levels on the cell surface. When differences in expression level were taken into account, Abruptex[M1] was comparable to Notch in its sensitivity to ligand-induced activation of reporter gene expression. Abruptex[M1] was also comparable to Notch in its requirement for modification by Fringe and in being sensitive to cis dowregulation by co-expressed ligands. By the available criteria Abruptex[M1] exhibits less activity than Notch. To explain the ectopic activity of Abruptex[M1] in vivo we suggest that it may be necessary to invoke an altered response to an as yet unidentified ligand or cofactor. PMID- 15804563 TI - Spatial arrangement of individual 4-cell stage blastomeres and the order in which they are generated correlate with blastocyst pattern in the mouse embryo. AB - In the unperturbed development of the mouse embryo one of the 2-cell blastomeres tends to contribute its progeny predominantly to the embryonic and the other to the abembryonic part of the blastocyst. However, a significant minority of embryos (20-30%) do not show this correlation. In this study, we have used non invasive lineage tracing to determine whether development of blastocyst pattern shows any correlation with the orientation and order of the second cleavage divisions that result in specific positioning of blastomeres at the 4-cell stage. Although the orientation and order of the second cleavages are not predetermined, in the great majority (80%) of embryos the spatial arrangement of 4-cell blastomeres is consistent with one of the second cleavages occurring meridionally and the other equatorially or obliquely with respect to the polar body. In such cleaving embryos, one of the 2-cell stage blastomeres tends to contribute to embryonic while the other contributes predominantly to abembryonic part of the blastocyst. Thus, in these embryos the outcome of the first cleavage tends to correlate with the orientation of the blastocyst embryonic-abembryonic axis. However, the order of blastomere divisions predicts a specific polarity for this axis only when the earlier 2-cell blastomere to divide does so meridionally. In contrast to the above two groups, in those embryos in which both second cleavage divisions occur in a similar orientation, either meridionally or equatorially, we do not observe any tendency for the 2-cell blastomeres to contribute to specific blastocyst parts. We find that all these groups of embryos develop to term with similar success, with the exception of those in which both second cleavage divisions occur equatorially whose development can be compromised. We conclude that the orientations and order of the second cleavages are not predetermined; they correlate with the development of blastocyst patterning; and that the majority, but not all, of these cleavage patterns allow equally successful development. PMID- 15804564 TI - EphB receptor tyrosine kinases control morphological development of the ventral midbrain. AB - EphB receptor tyrosine kinases and ephrin-B ligands regulate several types of cell-cell interactions during brain development, generally by modulating the cytoskeleton. EphB/ephrinB genes are expressed in the developing neural tube of early mouse embryos with distinct overlapping expression in the ventral midbrain. To test EphB function in midbrain development, mouse embryos compound homozygous for mutations in the EphB2 and EphB3 receptor genes were examined for early brain phenotypes. These mutants displayed a morphological defect in the ventral midbrain, specifically an expanded ventral midline evident by embryonic day E9.5 10.5, which formed an abnormal protrusion into the cephalic flexure. The affected area was comprised of cells that normally express EphB2 and ephrin-B3. A truncated EphB2 receptor caused a more severe phenotype than a null mutation, implying a dominant negative effect through interference with EphB forward (intracellular) signaling. In mutant embryos, the overall number, size, and identity of the ventral midbrain cells were unaltered. Therefore, the defect in ventral midline morphology in the EphB2;EphB3 compound mutant embryos appears to be caused by cellular changes that thin the tissue, forcing a protrusion of the ventral midline into the cephalic space. Our data suggests a role for EphB signaling in morphological organization of specific regions of the developing neural tube. PMID- 15804565 TI - Zebrafish pitx3 is necessary for normal lens and retinal development. AB - The human PITX3 gene encodes a bicoid-like homeodomain transcription factor associated with a variety of congenital ocular conditions, including anterior segment dysgenesis, Peter's anomaly, and cataracts. We identified a zebrafish pitx3 gene encoding a protein (Pitx3) that possesses 63% amino acid identity with human PITX3. The zebrafish pitx3 gene encompasses approximately 16.5kb on chromosome 13 and consists of four exons, which is similar to the genomic organization of other pitx genes. Expression of the zebrafish pitx3 gene was studied by in situ mRNA hybridization and RT-PCR. The pitx3 transcripts were detected throughout development with the greatest level of expression occurring in the developing lens and brain at 24hpf. In adults, the highest expression was detected in the eye. Morpholinos were used to knockdown expression of the Pitx3 protein and a control morpholino that contains five mismatched bases was used to confirm the specificity of the phenotypes. The morphants had small eyes, misshapen heads and reduced jaws and fins relative to controls. The morphants exhibited abnormalities in lens development and their retinas contained pyknotic nuclei accompanied by a reduction in the number of cells in different neuronal classes. This suggests the lens is required for retinal development or Pitx3 has an unexpected role in retinal cell differentiation or survival. These results demonstrate zebrafish pitx3 represents a true ortholog of the human PITX3 gene and the general function of the Pitx3 protein in lens development is conserved between mammals and the teleost fish. PMID- 15804566 TI - Functional analysis of seven genes encoding eight translation initiation factor 4E (eIF4E) isoforms in Drosophila. AB - The Drosophila genome-sequencing project has revealed a total of seven genes encoding eight eukaryotic initiation factor 4E (eIF4E) isoforms. Four of them (eIF4E-1,2, eIF4E-3, eIF4E-4 and eIF4E-5) share exon/intron structure in their carboxy-terminal part and form a cluster in the genome. All eIF4E isoforms bind to the cap (m7GpppN) structure. All of them, except eIF4E-6 and eIF4E-8 were able to interact with Drosophila eIF4G or eIF4E-binding protein (4E-BP). eIF4E-1, eIF4E-2, eIF4E-3, eIF4E-4 and eIF4E-7 rescued a yeast eIF4E-deficient mutant in vivo. Only eIF4E-1 mRNAs and, at a significantly lower level, eIF4E3 and eIF4E-8 are expressed in embryos and throughout the life cycle of the fly. The transcripts of the remaining isoforms were detected from the third instar larvae onwards. This indicates the cap-binding activity relies mostly on eIF4E-1 during embryogenesis. This agrees with the proteomic analysis of the eIF4F complex purified from embryos and with the rescue of l(3)67Af, an embryonic lethal mutant for the eIF4E-1,2 gene, by transgenic expression of eIF4E-1. Overexpression of eIF4E-1 in wild-type embryos and eye imaginal discs results in phenotypic defects in a dose-dependent manner. PMID- 15804567 TI - Pontin and Reptin regulate cell proliferation in early Xenopus embryos in collaboration with c-Myc and Miz-1. AB - Pontin (Tip49) and Reptin (Tip48) are highly conserved components of multimeric protein complexes important for chromatin remodelling and transcription. They interact with many different proteins including TATA box binding protein (TBP), beta-catenin and c-Myc and thus, potentially modulate different pathways. As antagonistic regulators of Wnt-signalling, they control wing development in Drosophila and heart growth in zebrafish. Here we show that the Xenopus xPontin and xReptin in conjunction with c-Myc regulate cell proliferation in early development. Overexpression of xPontin or xReptin results in increased mitoses and bending of embryos, which is mimicked by c-Myc overexpression. Furthermore, the knockdown of either xPontin or xReptin resulted in embryonic lethality at late gastrula stage, which is abrogated by the injection of c-Myc-RNA. The N termini of xPontin and xReptin, which mediate the mitogenic effect were mapped to contain c-Myc interaction domains. c-Myc protein promotes cell cycle progression either by transcriptional activation through the c-Myc/Max complex or by repression of cyclin dependent kinase inhibitors (p21, p15) through c-Myc/Miz-1 interaction. Importantly, xPontin and xReptin exert their mitogenic effect through the c-Myc/Miz-1 pathway as dominant negative Miz-1 and wild-type c-Myc but not a c-Myc mutant deficient in Miz-1 binding could rescue embryonic lethality. Finally, promoter reporter studies revealed that xPontin and xReptin but not the N-terminal deletion mutants enhance p21 repression by c-Myc. We conclude that xPontin and xReptin are essential genes regulating cell proliferation in early Xenopus embryogenesis through interaction with c-Myc. We propose a novel function of xPontin and xReptin as co-repressors in the c-Myc/Miz 1 pathway. PMID- 15804568 TI - Co-ordination of TGF-beta and FGF signaling pathways in bone organ cultures. AB - Transforming growth factor-beta (TGF-beta) is known to regulate chondrocyte proliferation and hypertrophic differentiation in embryonic bone cultures by a perichondrium dependent mechanism. To begin to determine which factors in the perichondrium mediate the effects of TGF-beta, we studied the effect of Insulin like Growth Factor-1 (IGF-I) and Fibroblast Growth Factors-2 and -18 (FGF2, FGF18) on metatarsal organ cultures. An increase in chondrocyte proliferation and hypertrophic differentiation was observed after treatment with IGF-I. A similar effect was seen after the perichondrium was stripped from the metatarsals suggesting IGF-I acts directly on the chondrocytes. Treatment with FGF-2 or FGF 18 resulted in a decrease in bone elongation as well as hypertrophic differentiation. Treatment also resulted in a decrease in BrdU incorporation into chondrocytes and an increase in BrdU incorporation in perichondrial cells, similar to what is seen after treatment with TGF-beta1. A similar effect was seen with FGF2 after the perichondrium was stripped suggesting that, unlike TGF-beta, FGF2 acts directly on chondrocytes to regulate proliferation and hypertrophic differentiation. To test the hypothesis that TGF-beta regulates IGF or FGF signaling, activation of the receptors was characterized after treatment with TGF beta. Activation was measured as the level of tyrosine phosphorylation on the receptor. Treatment with TGF-beta for 24h did not alter the level of IGFR-I tyrosine phosphorylation. In contrast, treatment with TGF-beta resulted in and increase in tyrosine phosphorylation on FGFR3 without alterations in total FGFR3 levels. TGF-beta also stimulated expression of FGF18 mRNA in the cultures and the effects of TGF-beta on metatarsal development were blocked or partially blocked by pretreatment with FGF signaling inhibitors. The results suggest a model in which FGF through FGFR3 mediates some of the effects of TGF-beta on embryonic bone formation. PMID- 15804569 TI - Six1 is not involved in limb tendon development, but is expressed in limb connective tissue under Shh regulation. AB - Mice deficient for the homeobox gene Six1 display defects in limb muscles consistent with the Six1 expression in myogenic cells. In addition to its myogenic expression domain, Six1 has been described as being located in digit tendons and as being associated with connective tissue patterning in mouse limbs. With the aim of determining a possible involvement of Six1 in tendon development, we have carefully characterised the non-myogenic expression domain of the Six1 gene in mouse and chick limbs. In contrast to previous reports, we found that this non-myogenic domain is distinct from tendon primordia and from tendons defined by scleraxis expression. The non-myogenic domain of Six1 expression establishes normally in the absence of muscle, in Pax3-/- mutant limbs. Moreover, the expression of scleraxis is not affected in early Six1-/- mutant limbs. We conclude that the expression of the Six1 gene is not related to tendons and that Six1, at least on its own, is not involved in limb tendon formation in vertebrates. Finally, we found that the posterior domain of Six1 in connective tissue is adjacent to that of the secreted factor Sonic hedgehog and that Sonic hedgehog is necessary and sufficient for Six1 expression in posterior limb regions. PMID- 15804570 TI - TGF-beta2 neutralization inhibits proliferation and activates apoptosis of cerebellar granule cell precurors in the developing cerebellum. AB - Transforming growth factor beta 2 (TGF-beta2) plays a critical role in growth, differentiation and cell death, but its function in the developing cerebellum is still uncertain. In this study we analyzed the effects of TGF-beta2 on ex vivo developing cerebellar slice cultures. Proliferation of granule cell precursors peaked ex vivo in the same developmental window as in vivo (P8-P14). Addition of recombinant TGF-beta2 could extent the proliferation of granule cell precursors and induced a second late proliferation wave. In contrast, antibody neutralization of TGF-beta2 strongly reduced proliferation and induced neurodegeneration. TGF-beta2 neutralization resulted in apoptotic cells, which showed caspase 3 activation. Taken together our results demonstrate that TGF beta2 is a novel growth and survival factor for granule cells precursors in the developing cerebellum. PMID- 15804572 TI - A hemocyte-specific integrin required for hemocytic encapsulation in the tobacco hornworm, Manduca sexta. AB - Upon encountering an object recognized as foreign, insect hemocytes aggregate in multiple layers on the surfaces of the object in a process known as encapsulation. For encapsulation to occur, hemocytes must switch from their usual nonadherent state to an adherent state, presumably by regulating the activity of adhesion proteins. Although detailed knowledge exists regarding the adhesion receptors for cells of the mammalian immune system, comparable information on adhesion molecules of insect hemocytes and their function in immune responses is extremely limited. We report here the identification of an integrin present exclusively on the surface of hemocytes in the tobacco hornworm, Manduca sexta. Monoclonal antibodies MS13 and MS34, which bind to plasmatocytes and block encapsulation, were used for immunoaffinity chromatography to isolate their corresponding hemocyte antigen, which was revealed to be the same integrin beta subunit. A cDNA for this M. sexta integrin beta1 was cloned and characterized. Integrin-beta1 mRNA was detected by Northern analysis in hemocytes and not in other tissues tested. MS13 and MS34 were demonstrated to bind to a recombinant fragment of integrin beta1 consisting of the I-like domain, consistent with their blocking of a ligand-binding site and subsequent disruption of plasmatocyte adhesion. Injection of double stranded integrin-beta1 RNA into larvae resulted in decreased integrin beta1 expression in plasmatocytes and significantly suppressed encapsulation. These results indicate that activation of ligand-binding by the hemocyte-specific integrin plays a key role in stimulating plasmatocyte adhesion leading to encapsulation. PMID- 15804571 TI - Altered BMP signaling disrupts chick diencephalic development. AB - The diencephalon is the caudal part of the forebrain and is organized into easily identifiable clusters of neurons called nuclei. Neurons in different nuclei project to discrete brain regions. Thus precise organization of the nuclei during forebrain development is necessary to build accurate neural circuits. How diencephalic development is regulated is poorly understood. BMP signaling participates in central nervous system patterning and development at many levels along the neural axis. Based on their expression we hypothesized BMPs play a role in diencephalic development. To test this hypothesis, we electroporated constitutively active and dominant negative forms of type I BMP receptors (Bmpr1a and Bmpr1b) into the embryonic chick forebrain. Ectopic induction of BMP signaling through constitutively active forms of the type I BMP receptors perturbs the normal gene expression patterns in the diencephalon and increases apoptotic cell death. These defects lead to disorganization of the diencephalic nuclei, suggesting BMP signaling is sufficient to modify diencephalic development. Loss-of-function studies, using dominant negative forms of Bmpr1a and Bmpr1b, indicate type I BMP receptors are necessary for normal eye and craniofacial development. However, they do not appear to be required for normal diencephalic development. In summary, our data indicate that while not necessary, BMP signaling via Bmpr1a and Bmpr1b, is sufficient to modify nuclear organization in the chick diencephalon. PMID- 15804573 TI - The emerging role of adenosine deaminases in insects. AB - Adenosine deaminases catalyze the deamination of adenosine and deoxyadenosine into their respective inosine nucleosides. Recent sequencing of the genomes of several model organisms and human reveal that Metazoa usually have more than one adenosine deaminase gene. A deficiency in the gene encoding the major enzyme is lethal in mouse and Drosophila and leads to severe combined deficiency (SCID) in human. In these organisms, enzyme deficiency causes increased adenosine/deoxyadenosine concentration in body fluids and some organs. Elevated levels of adenosine and deoxyadenosine are toxic to certain mammalian and insect cells, and it was shown for human and mouse that it is a primary cause of pathophysiological effects. Data suggest that the major role of adenosine deaminases in various taxa is the protection of tissues against increased levels of adenosine and deoxyadenosine. This review also discusses potential roles of adenosine deaminases in Drosophila metamorphosis and the employment of a Drosophila model to study the cell-specific toxicity of elevated nucleoside levels. PMID- 15804574 TI - Specific recognition, detoxification and metabolism of pyrrolizidine alkaloids by the polyphagous arctiid Estigmene acrea. AB - Evidence is presented that the polyphagous arctiid Estigmene acrea is well adapted to sequester and specifically handle pyrrolizidine alkaloids of almost all known structural types representative of the major plant families with pyrrolizidine alkaloid-containing species, i.e. Asteraceae with the tribes Senecioneae and Eupatorieae, Boraginaceae, Fabaceae, Apocynaceae and Orchidaceae. The adaptation of E. acrea to pyrrolizidine alkaloids includes a number of specialized characters: (i) highly sensitive recognition of alkaloid sources by pyrrolizidine alkaloid-specific taste receptors; (ii) detoxification of pyrrolizidine alkaloids by N-oxidation catalyzed by a specific flavin-dependent monooxygenase; (iii) transfer and maintenance of all types of pyrrolizidine N oxides through all developmental stages; (iv) conversion of the various structures into the male courtship pheromone hydroxydanaidal most probably through retronecine and insect specific retronecine esters (creatonotines) as common intermediates; (v) specific integration into mating behavior and defense strategies. Toxic otonecine derivatives, e.g. the senecionine analogue senkirkine, which often accompany the common retronecine derivatives and which cannot be detoxified by N-oxidation do not affect the development of E. acrea larvae. Senkirkine is not sequestered at all. Non-toxic 1,2-saturated platynecine derivatives that frequently occur together with toxic retronecine esters are sequestered and metabolized to hydroxydanaidal, indicating the ability of E. acrea to aromatize saturated pyrrolizidines. Although pyrrolizidine alkaloids, even if they are offered continuously at a high level (2%) in the larval diet, are non-toxic, E. acrea larvae are not able to develop exclusively on a pyrrolizidine alkaloid-containing plant like Crotalaria. Therefore, E. acrea appears to be specifically adapted to exploit pyrrolizidine alkaloid-containing plants as "drug source" but not as a food source. PMID- 15804575 TI - Increased expression of unusual EP repeat-containing proteins in the midgut of the tsetse fly (Glossina) after bacterial challenge. AB - Proteins containing a glutamic acid-proline (EP) repeat epitope were immunologically detected in midguts from eight species of Glossina (tsetse flies). The molecular masses of the tsetse EP proteins differed among species groups. The amino acid sequence of one of these proteins, from Glossina palpalis palpalis, was determined and compared to the sequence of a homologue, the tsetse midgut EP protein of Glossina m. morsitans. The extended EP repeat domains comprised between 36% (G. m. morsitans) and 46% (G. p. palpalis) of the amino acid residues, but otherwise the two polypeptide chains shared most of their sequences and predicted functional domains. The levels of expression of tsetse EP protein in adult teneral midguts were markedly higher than in midguts from larvae. The EP protein was detected by immunoblotting in the fat body, proventriculus and midgut, the known major immune tissues of tsetse and is likely secreted as it was also detected in hemolymph. The EP protein was not produced by the bacterial symbionts of tsetse midguts as determined by genome analysis of Wigglesworthia glossinidia and immunoblot analysis of Sodalis glossinidius. Bacterial challenge of G. m. morsitans, by injection of live E. coli, induced augmented expression of the tsetse EP protein. The presence of EP proteins in a wide variety of tsetse, their constitutive expression in adult fat body and midguts and their upregulation after immunogen challenge suggest they play an important role as a component of the immune system in tsetse. PMID- 15804576 TI - Identification of midgut proteins that are differentially expressed in trypanosome-susceptible and normal tsetse flies (Glossina morsitans morsitans). AB - Molecules in the midgut of tsetse flies (Diptera: Glossinidiae) are thought to play important roles in the life cycle of African trypanosomes by influencing initial parasite establishment and subsequent differentiation events that ultimately lead to maturation of mammal-infective trypanosomes. The molecular composition of the tsetse midgut is, therefore, of critical importance to disease transmission by these medically important vectors. In this study we compared protein expression profiles of midguts of the salmon mutant and wild type Glossina morsitans morsitans Westwood that display marked differences in their susceptibility to infection by African trypanosomes. Isotope coded affinity tag (ICAT) technology was used to identify 207 proteins including 17 that were up regulated and nine that were down regulated in the salmon mutants. Several of the up regulated molecules were previously described as tsetse midgut or salivary gland proteins. Of particular interest was the up regulation in the salmon flies of tsetse midgut EP protein, a recently described molecule with lectin-like activity that was also found to be induced in tsetse by bacterial challenge. The up regulation of the EP protein in midguts of salmon mutants was confirmed by two dimensional gel electrophoresis and tandem mass spectrometry. PMID- 15804577 TI - Characterization of a novel repetitive secretory protein specifically expressed in the modified salivary gland of Hydropsyche sp. (Trichoptera; Hydropsychidae). AB - Here, we report the cloning and characterization of a common salivary gland specific gene, Nf-1, from late instar Hydropsyche sp. larvae, and show that the corresponding gene product is translated in the gland and secreted to the gland lumen. The deduced Nf-1 protein is primarily composed of five repetitive sequence units of 63-65 amino acids, and contains a putative signal sequence composed of 19 amino acids. Secreted Nf-1 (approximately 37 kDa) was localized to the gland lumen by Western blotting of gland and lumen fractions. Together, the structure, expression pattern and protein localization of Nf-1 indicate that this protein is likely to be a major component of the silk shields and nets produced by the aquatic insect, Trichoptera. PMID- 15804578 TI - Melanogenesis and associated cytotoxic reactions: applications to insect innate immunity. AB - Insects transmit the causative agents for such debilitating diseases as malaria, lymphatic filariases, sleeping sickness, Chagas' disease, leishmaniasis, river blindness, Dengue, and yellow fever. The persistence of these diseases provides testimony to the genetic capacity of parasites to evolve strategies that ensure their successful development in two genetically diverse host species: insects and mammals. Current efforts to address the problems posed by insect-borne diseases benefit from a growing understanding of insect and mammalian immunity. Of considerable interest are recent genomic investigations that show several similarities in the innate immune effector responses and associated regulatory mechanisms manifested by insects and mammals. One notable exception, however, is the nearly universal presence of a brown-black pigment accompanying cellular innate immunity in insects. This response, which is unique to arthropods and certain other invertebrates, has focused attention on the elements involved in pigment synthesis as causing or contributing to the death of the parasite, and has even prompted speculation that the enzyme cascade mediating melanogenesis constitutes an ill-defined recognition mechanism. Experimental evidence defining the role of melanin and its precursors in insect innate immunity is severely lacking. A great deal of what is known about melanogenesis comes from studies of the process occurring in mammalian systems, where the pigment is synthesized by such diverse cells as those comprising portions of the skin, hair, inner ear, brain, and retinal epithelium. Fortunately, many of the components in the metabolic pathways leading to the formation of melanin have been found to be common to both insects and mammals. This review examines some of the factors that influence enzyme-mediated melanogenic responses, and how these responses likely contribute to blood cell-mediated, target-specific cytotoxicity in immune challenged insects. PMID- 15804579 TI - Activation of triacylglycerol lipase in the fat body of a beetle by adipokinetic hormone. AB - The activation of triacylglycerol lipase and the stimulation of proline synthesis in the fat body of the fruit beetle Pachnoda sinuata by the endogenous octapeptide hormone Melme-CC (pQLNYSPDWa), which belongs to the family of insect adipokinetic hormones, were studied, and the correlation of both events investigated. At rest, the activity of triacylglycerol lipase in the fat body of the beetle was higher than in the fat body of the American cockroach, Periplaneta americana, but lower than in the migratory locust, Locusta migratoria. Triacylglycerol lipase of the beetle is activated by: (a) injection of synthetic Melme-CC and (b) the stimulus of flight. Activation of lipase by Melme-CC is time dependent. Injection of cpt-cAMP activates triacylglycerol lipase in the fat body and causes an increase in the concentration of proline in the haemolymph at the expense of alanine. In contrast, injection of F-inositol-1,4,5-phosphate does not affect the activation state of lipase, nor the levels of amino acids in the haemolymph. High doses of octopamine do not activate lipase. Furthermore, activity of fat body lipase and proline concentration in the haemolymph both follow a circadian rhythm: both parameters are high in the morning, whereas they are low in the evening. When transfer of Melme-CC, released from the corpora cardiaca, to the thorax/abdomen is prevented by neck-ligation, the activity of lipase, as well as the circulating proline levels are low. Regression analysis revealed that activity of triacylglycerol lipase is positively correlated to proline concentration in the haemolymph, whereas there is a negative correlation of the enzyme activity and alanine level in the haemolymph. From these results we conclude that the activation of fat body triacylglycerol lipase by Melme-CC in P. sinuata stimulates proline synthesis. Proline is one of the major substrates to power flight activity in the beetle. PMID- 15804580 TI - Expression of genes encoding proteins involved in ecdysteroidogenesis in the female mosquito, Aedes aegypti. AB - A blood meal induces the ovaries of female Aedes aegypti mosquitoes to produce ecdysteroid hormones that regulate many processes required for egg maturation. Various proteins involved in the intracellular transport and biosynthesis of ecdysteroid precursors have been identified by analysis of Drosophila melanogaster mutants and by biochemical and molecular techniques in other insects. To begin examining these processes in mosquito ovaries, complete cDNAs were cloned for putative orthologs of diazepam-binding inhibitor (DBI), StAR related lipid transfer domain containing protein (Start1), aldo/keto reductase (A/KR), adrenodoxin reductase (AR), and the cytochrome P450 enzymes, CYP302a1 (22 hydroxylase), CYP315a1 (2-hydroxylase) and CYP314a1 (20-hydroxylase). As shown by RT-PCR, transcripts for all seven genes were present in ovaries and other tissues both before and following a blood meal. Expression of these genes likely supports the low level of ecdysteroids produced in vitro (7-10 pg /tissue/6 h) by tissues other than ovaries. Ovaries from females not blood fed and up to 6 h post blood meal (PBM) also produced low amounts of ecdysteroids in vitro, but by 18 and 30 h PBM, ecdysteroid production was greatly increased (75-106 pg/ovary pair/6h) and thereafter (48 and 72 h PBM) returned to low levels. As determined by real-time PCR analysis, gene transcript abundance for AedaeCYP302 and AedaeCYP315a1 was significantly greater (9 and 12 fold, respectively) in ovaries during peak ecdysteroid production relative to that in ovaries from females not blood fed or 2 h PBM. AedaeStart1, AedaeA/KR and AedaeAR also had high transcript levels in ovaries during peak ecdysteroid production, and AedaeDBI transcripts had the greatest increase at 48 h PBM. In contrast, gene transcript abundance of AedaeCYP314a1 decreased PBM. This study shows for the first time that transcription of a few key genes for proteins involved in ecdysteroid biosynthesis is positively correlated with the rise in ecdysteroid production by ovaries of a female insect. PMID- 15804581 TI - Ammonia metabolism in Aedes aegypti. AB - We investigated the mechanisms by which Aedes aegypti mosquitoes are able to metabolize ammonia. When females were given access to solutions containing NH(4)Cl or to a blood meal, hemolymph glutamine and proline concentrations increased markedly, indicating that ammonium/ammonia can be removed from the body through the synthesis of these two amino acids. The importance of glutamine synthetase was shown when an inhibitor of the enzyme was added to the meal causing the glutamine concentration in hemolymph to decrease significantly, while the proline concentration increased dramatically. Unexpectedly, we found an important role for glutamate synthase. When mosquitoes were fed azaserine, an inhibitor of glutamate synthase, the glutamine concentration increased and the proline concentration decreased significantly. This confirms the presence of glutamate synthase in mosquitoes and suggests that this enzyme contributes to the production of glutamate for proline synthesis. Several key enzymes related to ammonium/ammonia metabolism showed activity in homogenates of mosquito fat body and midgut. The mosquito genes encoding glutamate dehydrogenase, glutamine synthetase, glutamate synthase, pyrroline-5-carboxylate synthase were cloned and sequenced. The mRNA expression patterns of these genes were examined by a real time RT-PCR in fat body and midgut. The results show that female mosquitoes have evolved efficient mechanisms to detoxify large loads of ammonium/ammonia. PMID- 15804582 TI - Variants of the Drosophila melanogaster Ih-channel are generated by different splicing. AB - We isolated splice variants of the DMIH cDNA encoding members of the I(h)-channel family from Drosophila melanogaster by means of polymerase chain reaction and homology screening. Splicing at four different sites generates a great variety of different channel transcripts. The variants so obtained code for ion channel proteins with long or short N-termini and variations in the length of the interloop regions between the membrane-spanning domains S3-S4 and S4-S5. The multiple variants of DMIH coded by a single gene thus might form the molecular basis for a variety of I(h)-channels. Functional expression of one of the DMIH variants with a long N-terminus in HEK293 cells produced unitary currents that were preferentially selective for potassium over sodium ions and were activated by hyperpolarizing voltage steps. Cyclic nucleotides shifted the voltage activation curve to more positive membrane potentials. The current kinetics and modulatory influence of cyclic nucleotides resemble closely those of other invertebrate I(h)-channels, but activation by hyperpolarizing voltage steps had a V(1/2) of 123 mV, a more negative value than those of other recombinantly expressed insect I(h)-channels with a short N-terminus. PMID- 15804583 TI - The vistas and horizons of bony reconstruction--commentary. PMID- 15804584 TI - Mandibular defect repair by TGF-beta and IGF-1 released from a biodegradable osteoconductive hydrogel. AB - PURPOSE: Bone regeneration is crucial in the healing of defects in the craniofacial complex. The ability of transforming growth factor-beta1 (TGF-beta1) and insulin-like growth factor-1 (IGF-1), incorporated into a hydrogel scaffold to induce bone regeneration, was evaluated in a rat mandible defect model. MATERIAL AND METHODS: Hydrogel scaffolds containing either transforming growth factor-beta1 (TGF-beta1), insulin-like growth factor-1 (IGF-1), TGF-beta+ IGF-1, or saline, were implanted in rat mandibular bone defects. In a control group the defects were treated by saline alone. Bone defect healing was tested after 3 and 6 weeks by radiology and morphology. RESULTS: Soft tissue radiographs indicated that the area of new bone formation increased gradually after 3 and at 6 weeks. The percentage of closure after 3 weeks was less than the percentage closure after 6 weeks. The amount of calcified material in the TGF-beta and TGF-beta+IGF 1-treated groups had increased more than in the saline-containing hydrogel and control (saline-treated) defects. The percentages of defect closures were 37, 38, 24, 14, and 11% after 3 weeks, and 94, 91, 84, 72, and 29% after 6 weeks, in the TGF-beta+IGF-1, TGF-beta, IGF-1, saline containing hydrogel and saline-treated animals, respectively. Three-dimensional computerized tomography (3D CT) images showed that the 3D shape of the bones was restored. Morphological analysis of the defects treated with hydrogel containing TGF-beta, IGF-1 or TGF-beta+IGF-1 revealed significant bone formation after 6 weeks. CONCLUSION: It is concluded that the hydrogel scaffold impregnated with growth factors can induce bone regeneration and is therefore a promising surgical tool for enhancement of surgical repair of bone defects. PMID- 15804585 TI - An epidemiological study of orofacial clefts in Croatia 1988-1998. AB - INTRODUCTION: The objective of this epidemiological study was to assess the incidence at birth of orofacial clefts in Croatia. AIM: The aim of the present study was to analyse the character and incidence of orofacial clefts in Croatia and to compare the data with reports from other countries. MATERIAL AND METHODS: All the material for the epidemiological study was retrieved from the documented files from all the neonatal units and hospitals in Croatia providing surgical treatment. RESULTS: A total of 525,298 livebirths were documented during 11 years (1988-1998); 903 among them with orofacial clefts, 24 (2.7%) of them twins. Sixty (6.6%) infants died between birth and the age of 6 months. The incidence of orofacial clefts during the study period was 1.71 per thousand. When eliminating syndromic clefts, the incidence of non-syndromic clefts was 1.56 per thousand. Analysis of cleft lip with or without cleft palate (CL+/-P) and isolated cleft palate only (CP) revealed their incidence to be 1.05 and 0.66 per 1000, respectively. Of all types of clefting, CL and CLA was found in 17.2%, CL+/-P in 43.9%, CP in 38.2% and atypical facial clefts (AFC) in 0.8% of children. Left sided clefts were most common (51%), followed by bilateral (30.5%) and right sided (18.5%) clefts. The male to female ratio was 1.3. CL+/-P predominated in male and CP in female children. In 220 cases (24.4%) orofacial clefts were either associated with other anomalies or the clefts occurred as one feature of a syndrome. CONCLUSION: Data obtained from different sources yielded a cleft incidence of 1.71 per 1000 in Croatia. There were no differences in the incidences of orofacial clefts in comparison with similar data from other European countries. PMID- 15804586 TI - Non-syndromic orofacial clefts in Southern Italy: pattern analysis according to gender, history of maternal smoking, folic acid intake and familial diabetes. AB - BACKGROUND: Genetic studies have demonstrated that non-syndromic clefts of the lip, alveolus and palate have an heterogeneous genetic background, and that environmental factors contribute to the onset of this malformation. Therefore studies on different and homogeneous populations can be useful in detecting potentially related environmental and genetic factors. PURPOSE: The aim of the present study was to evaluate whether gender, folic acid intake, family history of diabetes and/or smoking during pregnancy were associated with a specific type of cleft in a group of patients affected by non-syndromic clefts, collected from Southern Italy. MATERIAL AND METHODS: Data from one hundred and twenty-six patients were evaluated retrospectively. Each cleft was described as composed by separate antomical entities such as lip, alveolus, primary and secondary palate. None had an isolated alveolar cleft and this was used as internal control. Pattern analysis was used to detect differences in the frequencies of any possible combination of 7 types of clefting stratified according to the studied variables. Data were analysed by comparing observed proportions. RESULTS: Isolated cleft palate as well as right-sided clefts of lip, alveolus and palate were more frequent in females (p = 0.0014 and 0.0281, respectively), while left sided clefts were more frequent in males (p = 0.0359). A lack of consumption of folic acid was associated with an higher incidence of clefts of the left lip (p = 0.018), while familial diabetes was associated more often with isolated cleft palate (p = 0.0014). CONCLUSIONS: Gender-related results were comparable with those found in Northern Italy and other countries. Environmentally related results disclosed specific subclasses of clefting associated with lack of folic acid consumption and familial diabetes. PMID- 15804587 TI - Self-reinforced biodegradable plates and screws for fixation of zygomatic fractures. AB - PURPOSE: The aim of this retrospective clinical study was to evaluate zygomatic fracture fixation with the BioSorbFX osteosynthesis system by assessing stability of reduction as well as complications in the first postoperative year and by conducting a survey to document surgeons' opinions on biodegradable osteosynthesis for this indication. MATERIAL: From January to September 2003, 25 patients with displaced non-infected unilateral fracture of the zygoma were operated upon (m:f = 20:5; age 17-81 years; mean 39.4 years) using the BioSorbFX 2.0 and/or 1.5 mm osteosynthesis systems. METHODS: A: Clinical and radiographic examinations were carried out immediately postoperatively and after 1, 3, 6, 9 and 12 months. METHODS: B: Surgeons were asked to participate in a survey critically evaluating their experience with biodegradable osteosynthesis systems. RESULTS: A: All fractures of the zygoma healed uneventfully. An excessive soft tissue reaction due to the degradation process was not seen. Three minor complications which resolved after local therapy occurred in the immediate postoperative phase. RESULTS: B: In the survey, the handling of biodegradable plates in general was rated worse than metal plate osteosynthesis. CONCLUSION: Fixation of fractures of the zygoma with the BioSorbFX system was simple and safe. The fixations remained stable and bony healing was uneventful. Postoperative complications were few, of a minor nature and not related to the process of biodegradation. PMID- 15804588 TI - Mandibular fractures in Jordanians: a comparative study between young and adult patients. AB - BACKGROUND: There are very few reports directly comparing mandibular fractures between young and adult patients in Middle Eastern countries. AIM: To review and compare the frequency and aetiology of mandibular fractures in young and adult Jordanian patients. STUDY DESIGN: A retrospective review was made of patients' records and radiographs seen at two university-based maxillofacial units during a 10-year period between 1993 and 2002. Age, gender, aetiology, and site were analysed and compared between young and adult patients. RESULTS: In total, 703 patients (502 (71%) male and 201 (29%) female) were found with 892 mandibular fractures altogether. There were 497 (71%) adults with 676 fractures, and 206 (29%) young patients with 216 fractures. The mean age of the adults was 30 years with a male-to-female ratio of 2.5:1. The mean age of the young patients was 13 years with a male-to-female ratio of 1.5:1. The most common fracture site in adult patients was the mandibular body, whereas the condyle predominated in young patients. The most common causative factor in adults was road traffic accidents, and falls in the young. CONCLUSION: There was a similar gender distribution in both groups of Jordanian young and adult patients. However, there were differences regarding aetiology and site of fracture. The differences found in this study between adult and young patients are similar to those from other countries. PMID- 15804589 TI - Changes in tongue and hyoid positions, and posterior airway space following mandibular setback surgery. AB - INTRODUCTION: The postural response of the tongue after mandibular setback is clinically important for maintaining normal respiration. Although the hyoid bone moves progressively to adapt physiologically to the altered orofacial configuration following such surgery, it is not clear whether repositioning of the hyoid has an effect on the pharyngeal airway. In the present study, postoperative changes in hyoid position and pharyngeal airway space were assessed retrospectively in patients who had undergone mandibular setback surgery. MATERIAL AND METHODS: Digitized lateral cephalograms from 30 mandibular setback surgery cases taken preoperatively, and 1 month and more than 1 year postoperatively, were used to examine pharyngeal airway morphology and the position of hyoid bone. RESULTS: A significant downward movement of the hyoid bone was found 1 month after surgery, while the pharyngeal airway dimensions at the tongue were maintained. More than 1 year after surgery, the hyoid position returned to its original position, resulting in a significant decrease in retrolingual airway dimension. CONCLUSION: The results indicate that mandibular setback causes airway narrowing late after surgery, while the early postoperative airway dimension is maintained. Long-term observations should be performed because of the changes of oropharyngeal configuration following mandibular setback. PMID- 15804590 TI - Long-term changes of hyoid bone position and pharyngeal airway size following mandibular setback by sagittal split ramus osteotomy. AB - INTRODUCTION: The purpose of this study was to determine long-term changes in hyoid bone position and pharyngeal airway size after mandibular setback osteotomies. MATERIAL AND METHODS: Serial cephalograms of 12 patients who underwent mandibular setback surgery were evaluated preoperatively, at 1 week, 6 months, and 14 months postoperatively, and finally after an average of 12 years. RESULTS: A mean setback of 5.6 mm was associated with a posterior and inferior movement of the hyoid bone. At long-term follow-up, the hyoid bone adapted horizontally to a position about 1.6 mm more posterior than its preoperative location. Immediately after surgery, the length of suprahyoid muscles correlated significantly with skeletal movements: suprahyoid muscles shortened by 4 mm but started lengthening after 1 year. Thereafter, it tended to correlate with the change of hyoid bone position rather than with skeletal relapse. Following the initial decrease after surgery, the size of the lower pharyngeal airway remained almost unchanged. The upper and middle pharyngeal airway sizes continued to decrease over the postoperative period of 12 years, and ultimately were smaller than their preoperative sizes by about 1.5 and 3 mm, respectively. PMID- 15804591 TI - Acupuncture analgesia and its application in cranio-maxillofacial surgical procedures. AB - BACKGROUND: The present study investigated the use of acupuncture analgesia in maxillofacial surgery. PATIENTS AND METHODS: Acupuncture analgesia was applied in 120 patients. In 20 of these, surgery was carried out under general anaesthesia in combination with acupuncture analgesia. In 100 patients, acupuncture analgesia was applied in addition to traditional postoperative analgesia. In case of troublesome postoperative pain, it was necessary to carry out additional sessions (1-4) of acupuncture stimulation. RESULTS: When acupuncture analgesia was used, the pulse rate and blood pressure during surgery generally remained stable. In the present study serum cortisol was also measured and showed only minor elevation. DISCUSSION: The present study demonstrated favourable results when the first variant of the brake method of acupoint stimulation was used for 40-50 min with additional manual twirling of the needles. This involved inserting needles into the acupoints and, using slow, rolling manual movements increasing the amplitude step-by-step provoking simultaneous especially strong patient sensations. CONCLUSION: Acupuncture analgesia can be a useful adjunct to conventional anaesthesia in maxillofacial surgery. PMID- 15804592 TI - Tape recorded hypnosis in oral and maxillofacial surgery--basics and first clinical experience. AB - BACKGROUND: Surgical treatment of diseases of the oral and maxillofacial region under local anaesthesia is quite commonly restricted by limited patient compliance. 'Medical Hypnosis' could be an alternative to treatment under pharmacological sedation. With this method, both autosuggestive and other suggestive procedures are used for anxiolysis, relaxation, sedation and analgesia of the patient. The purpose of this paper was to see whether there could be any potential for this treatment when operating on oral and maxillofacial patients. METHODS: During a 1-year-trial period, 209 operations under combined local anaesthesia/medical hypnosis were carried out on 174 non-preselected patients between the ages of 13 and 87 years. The surgical range covered oral, plastic and reconstructive, oncological, septic and trauma operations. RESULTS: Medical hypnosis turned out to be a reliable and standardizable method with high patient compliance. Remarkable improvements in treatment conditions for both patient and surgeons were achieved in 93% of cases. CONCLUSION: Controlled clinical studies are now necessary to obtain objective data on the effectiveness of hypnosis induced intraoperative effects in oral and maxillofacial surgery. PMID- 15804593 TI - Pneumosinus dilatans frontalis: two case reports. AB - INTRODUCTION: Pneumosinus dilatans was described first in 1898 by Meyes. Up to now only 60 cases have been published worldwide dealing mainly with young men between 20 and 30 years of age. The aetiology remains unknown. Whilst frontal sinuses are affected most often other sinuses can also be involved. The expansion of the pneumosinus is not only an aesthetic problem but may also have functional consequences. There may be a bulging of the roof of the orbit with consequent global displacement, ocular alterations and ptosis. Other features are headache and anosmia. PATIENTS: Two male patients with a pneumosinus dilatans of the frontal sinus are presented. They were 22 and 25 years old. The main complaints were aesthetic disfigurement. Treatment consisted of craniofacial repair of the malformation. RESULTS: While one patient was lost to follow-up, the other still is happy and has a stable aesthetic long-term result. PMID- 15804594 TI - A new retractor for surgical removal of mandibular torus. AB - INTRODUCTION: A newly devised retractor for mandibular torus is presented. It has a half-spoon-shaped head and was applied to various cases of mandibular torus for its evaluation. RESULTS: It was found that the elevated mucoperiosteum flap was safely protected lingually by using this retractor. Various mandibular tori were completely resected without damaging the surrounding tissues. PMID- 15804595 TI - Differential inflammatory activation of IL-6 (-/-) astrocytes. AB - IL-6 is a major immunomodulatory cytokine with neuroprotective activity. The absence of interleukin-6 (IL-6) results in increased vulnerability of dopaminergic neurons to the neurotoxicant, MPTP, and a compromised reactive microgliosis. To determine how astrogliosis may contribute to nigrostriatal degeneration in IL-6 (-/-) mice, the inflammatory profiles of astrocytes of IL-6 genotype were compared. Fourteen cytokines and four chemokines were simultaneously assayed in the supernatants of LPS-stimulated primary astrocyte cultures. In a time course of 6, 18 and 48 h and LPS stimulations of 0, 0.1, 1, 10 and 100 ng/ml, IL-6 (-/-) astrocytes secreted significantly greater amounts of the pro-inflammatory cytokines IL-1alpha, IL-1beta and TNFalpha than did IL-6 (+/+) cells. Elevated levels of IL-10 and IL-12p40 were only detected at 48 h post-stimulation with greater IL-10 in IL-6 (-/-) supernatants and greater IL 12p40 in IL-6 (+/+) supernatants. IL-6 (+/+) astrocytes produced more G-CSF and GM-CSF when compared with IL-6 (-/-) astrocytes. Chemokine levels were greater in supernatants of IL-6 (+/+) astrocytes than IL-6 (-/-) cells prior to 48 h post stimulation. At that time, higher levels of MIP-1alpha were maintained in IL-6 (+/+) supernatant, while similar levels of MCP-1 in supernatants of both IL-6 (+/+) and IL-6 (-/-) cells were measured. Additionally, LPS (100 ng/ml) resulted in greater levels of KC and Rantes in IL-6 (-/-) astrocyte supernatants compared with IL-6 (+/+) supernatants at that time. These results suggest that the autocrine modulatory activities of IL-6 affect multiple cytokine secretory pathways, which could participate in neurodegenerative processes. PMID- 15804596 TI - Cytokine responses of human gingival fibroblasts to Actinobacillus actinomycetemcomitans cytolethal distending toxin. AB - Actinobacillus actinomycetemcomitans is implicated in the pathogenesis of localized aggressive periodontitis, and has the capacity to express a cytolethal distending toxin (Cdt). Gingival fibroblasts (GF) are resident cells of the periodontium, which can express several osteolytic cytokines. The aims of this study were a) to investigate the role of Cdt in A. actinomycetemcomitans-induced expression of osteolytic cytokines and their cognate receptors in GF and b) to determine if the previously demonstrated induction of receptor activator of NFkappaB ligand (RANKL) by A. actinomycetemcomitans is mediated by these pro inflammatory cytokines or by prostaglandin E(2) (PGE(2)). A. actinomycetemcomitans clearly induced interleukin (IL)-6, IL-1beta, and to a minimal extent, tumor necrosis factor (TNF)-alpha mRNA expression. At the protein level, IL-6 but not IL-1beta or TNF-alpha expression was stimulated. The mRNA expression of the different receptor subtypes recognizing IL-6, IL-1beta and TNF alpha was not affected. A cdt-knockout strain of A. actinomycetemcomitans had similar effects on cytokine and cytokine receptor mRNA expression, compared to its parental wild-type strain. Purified Cdt stimulated IL-6, but not IL-1beta or TNF-alpha protein biosynthesis. Antibodies neutralizing IL-6, IL-1 or TNF-alpha, and the PGE(2) synthesis inhibitor indomethacin, did not affect A. actinomycetemcomitans-induced RANKL expression. In conclusion, a) A. actinomycetemcomitans induces IL-6 production in GF by a mechanism largely independent of its Cdt and b) A. actinomycetemcomitans-induced RANKL expression in GF occurs independently of IL-1, IL-6, TNF-alpha, or PGE(2). PMID- 15804597 TI - The effect of MCP-1 depletion on chemokine and chemokine-related gene expression: evidence for a complex network in acute inflammation. AB - The expression of chemokines has been suggested to involve an interdependent network, with the absence of a single chemokine affecting the expression of multiple other chemokines. Monocyte chemoattractant protein (MCP-1), a member of C-C chemokine superfamily, plays a critical role in the recruitment and activation of leukocytes during acute inflammation. To examine the effect of the loss of MCP-1 on expression of the chemokine network, we compared the mRNA expression profiles of MCP-1(-/-) and wild type mice during the acute inflammatory phase of excisional wounds. Utilizing a mouse cDNA array containing 514 chemokine and chemokine related genes, the loss of MCP-1 was observed to cause a significant upregulation of nine genes (Decorin, Persephin, IL-1beta, MIP 2, MSP, IL1ra, CCR5, CCR3, IL-11) and significant downregulation of two genes (CCR4 and CD3Z) in acute wounds. The array data was confirmed by semi quantitative RT-PCR. The effect of MCP-1 deletion on chemokine expression was further examined in isolated macrophages. Compared to wild type, LPS-stimulated peritoneal macrophages from MCP-1(-/-) mice showed a significant increase in the expression of RANTES, MIP-1beta, MIP-1alpha and MIP-2 mRNA. The data suggest that loss of a single chemokine perturbs the chemokine network not only in the setting of acute inflammation but even in an isolated inflammatory cell, the macrophage. PMID- 15804598 TI - Cytokine production of stimulated whole blood cultures in rheumatoid arthritis patients receiving short-term infliximab therapy. AB - Patients with rheumatoid arthritis (RA) treated with anti-tumor necrosis factor (TNF) strategies have an increased susceptibility to infections, especially those caused by intracellular pathogens. In this study we assessed the cytokine production capacity in patients with RA and we further investigated whether anti TNF therapy modulates the production of pro-inflammatory cytokines involved in the resistance against infections. Whole blood cultures from 10 RA patients and 10 healthy controls were stimulated with heat-killed Candida albicans, Salmonella typhimurium, Staphyloccocus aureus, Aspergillus fumigatus or Mycobacterium tuberculosis and production of interleukin (IL)-1beta, IL-6, IL-10, interferon (IFN)-gamma and TNF-alpha was measured. Before anti-TNF therapy, whole blood cultures from RA patients released significantly less IFN-gamma than healthy controls after stimulation with all tested microorganisms. Short-term anti-TNF therapy did not have an inhibitory effect on the release of the cytokines tested. We conclude that cells of patients with RA have a strongly reduced production capacity of IFN-gamma after bacterial challenge. Although short-term therapy with anti-TNF agents did not further decrease the release of other proinflammatory cytokines, the combination of defective IFN-gamma production in basal conditions and TNF neutralization during anti-TNF therapy is likely to be responsible for the higher susceptibility to infections in patients with RA. PMID- 15804599 TI - Recombinant rat IL-1beta and IL-6 synergistically enhance C3 mRNA levels and complement component C3 secretion by H-35 rat hepatoma cells. AB - Hepatic synthesis of complement component C3 is regulated in part by inflammatory cytokines. Rat models are frequently employed to investigate pathogenic roles of complement and cytokines. However, cytokines obtained from species other than the rat were used in previous studies of cytokine regulation of C3 synthesis in rat hepatocytes or hepatoma cells. It is not known whether these prior reports predict hepatocellular responses evoked by rat cytokines. Therefore, H-35 rat hepatoma cells were employed to measure the effect of recombinant rat IL-1beta, IL-6, IFN-gamma, and TNF-alpha on C3 protein secretion and C3 mRNA levels quantified by ELISA and quantitative RT-PCR. Compared to untreated control cells, H-35 cells treated with IL-1beta, IL-6, and IFN-gamma increased C3 secretion approximately 10-, 4-, and 2-fold, respectively. TNF-alpha was toxic, precluding further analysis. IL-1beta and IL-6 demonstrated synergy with respect to the quantity and rate of increase of C3 mRNA measured and the magnitude of C3 protein secretion. Previous reports using non-rat cytokines did not consistently predict H-35 responses to rat cytokines. Consequently, we recommend the use of rat cytokines in rat models that include analysis of cytokine-mediated events. PMID- 15804600 TI - Interferon-gamma associated cytokines and chemokines produced by spleen cells from Brucella-immune mice. AB - It is known that interferon (IFN)-gamma plays a critical role in protection against brucellosis. In this study we have investigated several cytokines and chemokines that are associated with IFN-gamma for potential in vitro correlates of protection. We cultured spleen cells in vitro from mice immunized orally with a live, attenuated Brucella melitensis vaccine candidate (WR201) and stimulated these cells with a lysate of B. melitensis. Differential gene expression of several cytokines and chemokines in stimulated spleen cells was analysed by real time PCR, and secreted proteins were determined by ELISA. Immunized mice produced higher levels of both protein and gene transcripts for IFN-gamma, interleukin (IL)-2, IL-18 and MIP1-alpha. Immunized mice also had elevated gene expression levels for IL12-p40, IL23-p19, IP-10, MIG and MCP-1 when compared to normal mice. In this study we have identified new cytokines and chemokines as potential immune correlates in responses to protection in Brucella-vaccinated mice. PMID- 15804602 TI - Assay for the transbilayer movement of polyisoprenoid-linked saccharides based on the transport of water-soluble analogues. AB - Flippases are a class of membrane proteins that are proposed to facilitate the transbilayer movement of amphipathic polar lipids that are required for membrane biogenesis and the assembly of many diverse complex glycoconjugates in eukaryotic and prokaryotic cells. Despite their crucial roles in membrane biology, very little is known about their structures and the precise mechanism(s) by which they overcome the biophysical barriers of the hydrophobic core, and allow polar head groups to traverse membrane bilayers. This chapter presents methods based on the transport of water-soluble analogues that can be applied to investigate membrane proteins mediating the transverse diffusion of polyisoprenoid-linked glycolipid intermediates involved in the biosynthesis of N-linked glycoproteins, glycosylphosphatidylinositol anchors and bacterial polysaccharides. PMID- 15804603 TI - Fluorophore-assisted carbohydrate electrophoresis: a sensitive and accurate method for the direct analysis of dolichol pyrophosphate-linked oligosaccharides in cell cultures and tissues. AB - Lipid-linked oligosaccharides (LLOs) such as Glc3Man9GlcNAc2-P-P-dolichol are the precursors of asparagine (N)-linked glycans, which are essential information carriers in many biological systems, and defects in LLO synthesis cause Type I Congenital Disorders of Glycosylation. Due to the low abundance of LLOs and the limitations of the chemical and physical methods previously used to detect them, almost all studies of LLO synthesis have relied upon metabolic labeling of the oligosaccharides with radioactive sugar precursors such as [3H]mannose or [14C]glucosamine. In this article, a procedure is presented for a facile, accurate, and sensitive non-radioactive method for LLO analysis based on fluorophore-assisted carbohydrate electrophoresis (FACE). First, LLOs are extracted and partially purified. Next, oligosaccharides released from LLOs are labeled with negatively charged fluorophores: 8-aminonaphthalene-1,3,6 trisulfonate (ANTS) or 7-amino-1,3-naphthalenedisulfonic acid (ANDS). A specialized form of polyacrylamide gel electrophoresis is then used to resolve and measure ANTS or ANDS labeled oligosaccharides. Finally, the resolved oligosaccharides are detected and quantified by fluorescence imagers using CCD cameras. PMID- 15804604 TI - Glycoprotein reglucosylation. AB - Proteins following the secretory pathway acquire their proper tertiary and in certain cases also quaternary structures in the endoplasmic reticulum (ER). Incompletely folded species are retained in the ER and eventually degraded. One of the molecular mechanisms by which cells achieve this conformational sorting is based on monoglucosylated N-glycans (Glc1Man5-9GlcNAc2) present on nascent glycoproteins in the ER. This chapter discusses two of the steps that regulate the abundance of such N-glycan structures, including glycoprotein deglucosylation (by glucosidase II) and reglucosylation (by the UDP-Glc:glycoprotein glucosyltransferase), as well as an overview of methods to evaluate the N-glycans prevalent during glycoprotein biogenesis in the ER. PMID- 15804605 TI - In vitro and in vivo assays to assess the functions of calnexin and calreticulin in ER protein folding and quality control. AB - Newly synthesized polypeptides entering the endoplasmic reticulum (ER) encounter a large array of molecular chaperones and folding factors that facilitate proper folding as well as assess folding status, retaining non-native proteins within the ER. Calnexin (CNX), an ER membrane protein, and its soluble homologue, calreticulin (CRT), are two important molecular chaperones that contribute to both processes. They are highly unusual chaperones in that they act as lectins, binding the Asn-linked oligosaccharides of newly synthesized glycoproteins, as well as recognizing the polypeptide segments of glycoproteins. Furthermore, they associate with ERp57, a thiol oxidoreductase, that is thought to enhance the oxidative folding of glycoproteins bound to CNX/CRT. These characteristics of CNX and CRT as well as their mode of action have been elucidated though the use of multiple in vitro and in vivo approaches. This chapter will focus on the description of a number of in vitro assays that have been used to characterize the lectin and ERp57-binding functions of CNX/CRT and also their abilities to act as molecular chaperones to suppress protein aggregation. In addition, we will describe insect and mammalian expression systems in which major histocompatibility complex class I molecules are used as model glycoprotein substrates for CNX and CRT. These systems have been valuable in assessing folding and quality control events in vivo that are influenced by CNX or CRT as well as in characterizing the spectrum of substrates that are recognized by these chaperones. PMID- 15804606 TI - Alpha1-antitrypsin as model to assess glycan function in endoplasmic reticulum. AB - It is now understood that a cohesive series of quality control checkpoints ensures the accuracy of gene expression in eukaryotic cells. Although initiated in the nucleus to monitor the integrity of inherited genetic information, the quality control program encompasses post-translational events that facilitate the structural maturation of encoded proteins or target them for degradation if unable to adopt native structure. Given the fact that many genetic mutations actually manifest themselves at the level of aberrant protein structure, a current challenge in the post-genomics era is to elucidate how post-translational checkpoints can modify the severity of numerous loss-of-function and gain-of toxic-function diseases, possibly influencing an individual's susceptibility toward the development of the associated pathologies. The purpose of this chapter is to describe the experimental methodology by which alpha1-antitrypsin has been used as a molecular reagent to define the mechanisms by which the processing and recognition of asparagine-linked oligosaccharides can orchestrate the fate of newly synthesized glycoproteins in the early secretory pathway. The conceptual framework, and associated techniques, can serve as a roadmap for the investigation of other mutated glycoproteins, many of which can contribute to disease. PMID- 15804607 TI - An in vitro assay for the selective endoplasmic reticulum associated degradation of an unglycosylated secreted protein. AB - The endoplasmic reticulum (ER) represents the first compartment into which nascent secreted proteins traffic, and not coincidentally the ER lumen houses a high concentration of factors that facilitate protein folding, such as molecular chaperones. To off-set the potentially lethal consequences of mis-folded secreted protein accumulation, aberrant proteins may be selected for degradation via a process known as ER associated degradation (ERAD). After their selection ERAD substrates are retro-translocated back to the cytoplasm and then degraded by the 26S proteasome. Key features of the selection, retro-translocation, and degradation steps that constitute the ERAD pathway were elucidated through the development of an in vitro ERAD assay. In this assay the fates of two yeast proteins can be distinguished after their translocation, or import into ER derived microsomes. Whereas a wild type, glycosylated protein ("Gp(alpha)F") is stable, a non-glycosylated version of the same protein ("p(alpha)F") is rapidly degraded when microsomes containing radiolabeled forms of these substrates are incubated in cytosol and ATP. The purpose of this chapter is first to discuss the experimental findings from the use of the in vitro assay, and then to describe the assay in detail. Finally, future potential uses of the in vitro system are illustrated. PMID- 15804608 TI - A simple, sensitive in vitro assay for cytoplasmic deglycosylation by peptide: N glycanase. AB - A cytoplasmic peptide: N-glycanase (PNGase) has been implicated in the proteasomal degradation of aberrant glycoproteins synthesized in the endoplasmic reticulum. The reaction is believed to be important for subsequent proteolysis by the proteasome since bulky N-glycan chains on misfolded glycoproteins may impair their efficient entry into the interior of the cylinder-shaped 20S proteasome, where its active site resides. This cytoplasmic enzyme was first detected in 1993 by a simple, sensitive assay method using 14C-labeled glycopeptide as a substrate. The deglycosylation reaction by PNGase brings about two major changes on substrate the peptide; one is removal of the N-glycan chain and the other is the introduction of a negative charge into the core peptide by converting the glycosylated asparagine residue(s) into an aspartic acid residue(s). The assay method we developed monitors these major changes in the core peptide, and the respective changes were detected by distinct analytical methods: i.e., paper chromatography and paper electrophoresis. This chapter will describe the simple, sensitive in vitro assay method for PNGase. PMID- 15804609 TI - Screening for mutants defective in secretory protein maturation and ER quality control. AB - A genetic strategy devised to understand the physiology of the unfolded protein response serendipitously generated mutants affecting a broad spectrum of functions needed for secretory protein biogenesis and quality control. These included N- and O-linked glycosylation, glycosylphosphatidylinositol anchor biosynthesis and transfer, protein folding, protein trafficking, lumenal ionic homeostasis, ER quality control, and ER associated protein degradation. As these pathways are incompletely understood, the screen provides a simple method for their genetic dissection. This article describes methods for isolating novel mutants of these pathways and strategies for identifying corresponding genes. PMID- 15804610 TI - The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress. AB - The multiple implications of ER stress and the unfolded protein response in health and disease highlight the importance of identifying convenient monitoring systems for its onset under various experimental or physiological settings. A large volume of studies establish that induction of GRP78 is a marker for ER stress. GRP78, also referred to as BiP, is a central regulator for ER stress due to its role as a major ER chaperone with anti-apoptotic properties as well as its ability to control the activation of transmembrane ER stress sensors (IRE1, PERK, and ATF6) through a binding-release mechanism. In the following report, we present several methods to measure GRP78 induction. This can be achieved by measuring the Grp78 promoter activity or by measuring the level of Grp78 transcripts or GRP78 protein. These techniques can be applied to tissue culture cells as well as tissues and organs. PMID- 15804611 TI - ER stress signaling by regulated proteolysis of ATF6. AB - ATF6 is an endoplasmic reticulum (ER) membrane-anchored transcription factor activated by intramembrane proteolysis in the ER stress response. Upon ER stress, ATF6 is transported from the ER to the Golgi to be processed by site-1 and site-2 proteases. The trafficking is controlled by the ER chaperone BiP/GRP78. Here, we describe the experimental methods that we have used to study of ATF6 regulation in tissue culture cells. These methods were used to investigate several key steps of ATF6 activation in the ER stress response including binding and dissociation of BiP to ATF6, translocation from the ER to the Golgi and cleavage in the Golgi. In addition, luciferase reporter assays were a sensitive way to monitor ER stress and ATF6 activation. These methods were not only useful for the study ATF6 and the ER stress response, they might also help to elucidate the roles of the ER stress response in a number of human diseases involving misfolded proteins and in the differentiation of secretory tissues which require higher ER folding capacities. PMID- 15804612 TI - Quantitative measurement of events in the mammalian unfolded protein response. AB - The accumulation of unfolded or misfolded proteins in the endoplasmic reticulum (ER) triggers the unfolded protein response. IRE1, PERK, ATF6, BiP, EDEM, lipid linked oligosaccharides (LLOs), and XBP1 directly or indirectly participate in this process. This article provides methods used in our laboratory to quantitatively measure the accumulation of mRNAs encoding BiP and EDEM; splicing of XBP-1; cleavage of ATF6; inhibition of protein synthesis by PERK; and extension of LLOs under control and stress conditions. PMID- 15804613 TI - ER stress signaling by regulated splicing: IRE1/HAC1/XBP1. AB - The endoplasmic reticulum (ER) serves many specialized functions in the cell including calcium storage and gated release, biosynthesis of membrane and secretory proteins, and production of lipids and sterols. Therefore, the ER integrates many internal and external signals to coordinate downstream responses, although the mechanism(s) that maintain homeostasis are largely unknown. When misfolded or unfolded proteins accumulate in the ER, an intracellular signaling pathway termed the unfolded protein response (UPR) is activated. Identification of IRE1 in the yeast Saccharomyces cerevisiae as a proximal sensor in the UPR pathway was a milestone in understanding how the ER responds to the accumulation of unfolded protein and signals transcriptional activation through regulated nonconventional splicing of its substrate mRNA encoding the transcription factor Hac1p. Subsequent studies identified IRE1 and HAC1 homologues in mammalian cells. Here, we summarize various approaches to study the IRE1-Hac1 pathway in yeast and the homologous IRE1-XBP1 pathway in mammalian cells. We present microbiological growth assays for the UPR, reporter assays for UPR signaling, direct techniques to measure UPR activation in vivo, methods to study translation of HAC1 mRNA, and in vitro cleavage and ligation of HAC1 and XBP1 mRNA. Especially we think the newly developed quantitative and qualitative methods to detect IRE1 activity dependent XBP1 mRNA splicing will be fast and accurate tools to show the activation of the UPR. PMID- 15804614 TI - [Epidemiology of head and neck carcinomas]. AB - Head and neck carcinomas, i.e. oral cavity, pharynx and larynx cancers, are frequent among men in France. They are mostly squamous-cell carcinomas. The main risk factors are alcohol and tobacco, and their effects are multiplicative. In France, alcohol and tobacco consumptions are decreasing in the male population leading to decrease in head and neck carcinoma incidence and mortality. The best way to prevent head and neck carcinoma is the reduction of alcohol and tobacco consumption. PMID- 15804615 TI - [Current applications and future developments of positron emission tomography in head and neck cancer]. AB - Positron emission tomography (PET-scan) is a well-established imaging modality in oncology. Using FDG, PET has also a wide range of applications in head and neck tumors for diagnosis, staging, monitoring of response to therapy, and detection of relapse. After a short technical introduction, the current indications of PET FDG in head and neck tumors are reviewed. Present and future developments of PET are twofold: the use of new tracers for protein synthesis, cellular proliferation or detection of hypoxia etc., and the introduction of metabolic imaging as a adjunct to CT and MRI to determine target-volumes in radiation treatment planning. However, it has to be emphasized that a thorough clinical validation of the methods used is mandatory before their implementation in routine practice. PMID- 15804616 TI - [Reconstructive surgery in head and neck oncology: indication and technic]. AB - Oncologic cervicofacial surgery and plastic surgery have had a common evolution over the last 50 years where progress erasing from one was beneficial to the other one. We review here the historical evolution of these specialties and present the state of the art of plastic surgery in the field of cervicofacial oncology. PMID- 15804617 TI - [Individual patient data meta-analyses in head and neck carcinoma: what have we learnt?]. AB - Carcinoma of the upper aerodigestive tract (oral cavity, oropharynx, hypopharynx, nasopharynx, larynx) are frequent tumors for which surgery and/or radiotherapy are the main therapeutic agents. The main results of meta-analyses based on the collection of individual patients data are reported: 1) The meta-analysis on chemotherapy, regrouping data of nearly 11,000 patients issued from 63 randomized trials showed an absolute benefit of 4% at five years in overall survival, in favor of chemotherapy (P<0.0001). Most of the benefit was seen with concomitant radiochemotherapy, however with a relatively large heterogeneity in this subgroup of trials. An update of this meta-analysis was performed including 24 additional trials, which confirmed the magnitude of the benefit due to concomitant chemotherapy (8% at 5 years). 2) The meta-analysis on larynx preservation, using induction chemotherapy in larynx and hypopharynx carcinomas. No significant difference was seen between the control arm with total laryngectomy and the larynx preservation approach. 3) The meta-analysis on chemotherapy in nasopharynx carcinomas, from the data of 11 randomized trials including 2722 patients, and comparing the radiotherapy to radio-chemotherapy (1979-2001). The results showed an absolute benefit of 6% at five years in overall survival, in favor of chemotherapy (P<0.0001). Most of the benefit was seen with concomitant radiochemotherapy. 4) Finally, a meta-analysis on altered fractionated RT, compared to conventional RT in 15 randomized trials regrouping 6515 patients. The results showed a small but significant improvement in favor of altered fractionated RT for overall survival and local control with an absolute benefit at five years of 3 and 6%, respectively. PMID- 15804618 TI - [Larynx preservation, state of the art]. AB - Larynx preservation in advanced pharyngolaryngeal cancers has been a major challenge in clinical research over the past two decades. Subtotal surgery (endoscopic laser surgery, supracricoid partial laryngectomies and hemilaryngopharyngectomies) has allowed reducing the indications of the so-called "mutilating" surgery. On the other hand, the modification of fractionation has notably improved the results of definitive irradiation (but most probably for supraglottic tumors). The main clinical research has been carried out with either sequential or concurrent chemo-irradiation. Induction chemotherapy followed by irradiation in good responders or by surgery in poor responders allowed to preserve the larynx in around half the cases without deleterious impact on overall survival. Concurrent chemo-irradiation trials suggested that the larynx preservation rates could be increased but overall survival remained unchanged and mucositis was a notable side effect. The next step could be the combination of induction chemotherapy followed by concurrent chemo-irradiation in good responders. Finally, the place of new drugs (taxanes, targeted therapies) is to be explored in this context. PMID- 15804619 TI - Intensity modulated radiation therapy in head and neck squamous cell carcinoma: state of the art and future challenges. AB - Intensity-modulated radiation therapy (IMRT) for head and neck (HN) tumors refers to a new approach to the whole treatment procedure from patient immobilization to beam delivery. Implementation of IMRT thus requires knowledge of setup uncertainties, adequate selection and delineation of target volumes based on clinical examination and optimal imaging modalities, appropriate specification and dose prescription regarding dose-volume constraints, and ad hoc quality control of both the clinical and physical aspects of the whole procedure. A large number of issues still need to be resolved and/or further refined, such as the optimal selection and delineation of the target volume in particular, with the introduction of functional imaging, and a better integration of improved dose distribution into the fractionation strategy. IMRT is associated with a potentially increased incidence of carcinogenesis, although in the HN area this risk is relative to the intrinsic risk of co-morbidity and secondary cancer associated with the patient's lifestyle. Currently, the implementation of IMRT into routine clinical practice for HN cancers may not be a straightforward matter, and should probably be restricted to selected patients and selected institutions with adequate resources and experience. This review emphasizes the above aspects and provides some recommendations for the future use of IMRT in patients with HN tumors. PMID- 15804620 TI - [EGF receptor in head and neck cancer]. AB - EGFR is overexpressed and is associated with a poor prognosis in head and neck cancer. Among the biological and cellular effects resulting from EGFR targeting in head and neck cancer there is the capacity to restore apoptotic capacities. Other experimental results put into evidence that DNA-repair activity was reduced by the application of EGFR targeting agents. This context was in favor of a research oriented towards combination between anti-EGFR drugs and cytotoxic agents, particularly irradiation. Supra-additive cytotoxic effects have been observed at the experimental level when combining anti-EGFR drugs with irradiation in head and neck cancer. These experimental data were recently confirmed at the clinical level in locally advanced head and neck cancer. PMID- 15804621 TI - [Nasopharyngeal carcinomas: from biology to clinic]. AB - Nasopharyngeal carcinomas (NPC) are very different from other head and neck cancers because of their specific multifactorial etiology and their geographic distribution. Epstein-Barr Virus (EBV) is implicated in oncogenesis of NPC in association with genetic alterations such as inactivation of the p16/Ink4, p19/ARF, RASSF1 or Blu genes. Tumoral tissues include a very abundant characteristic lymphoid infiltrate. Inflammatory cytokines are produced by both malignant and infiltrating cells. There is no efficient immune response against the tumor. On the opposite, infiltrating lymphocytes might play a role in tumor development. Serological methods and detection of circulating viral DNA are expected to become useful for early detection of relapse and on a longer term for primary screening. NPC are often diagnosed at a late stage because patients may remain asymptomatic for a long time. Computed tomography (CT scan) and magnetic resonance imaging (MRI) are complementary for the initial evaluation. Positron emission tomography (PET) is efficient for the evaluation of treatment efficiency and detection of relapses. Treatment is based on radiotherapy and chemotherapy. Their optimal use needs to be evaluated by phase III trials but positive results have been obtained by concomitant association of radiotherapy and chemotherapy. Targeted therapies are being studied with strategies based on disruption of viral latency, use of replicative adenoviruses or anti-tumor vaccination. PMID- 15804623 TI - Relating structure to mechanism in creatine kinase. AB - Found in all vertebrates, creatine kinase catalyzes the reversible reaction of creatine and ATP forming phosphocreatine and ADP. Phosphocreatine may be viewed as a reservoir of "high-energy phosphate" which is able to supply ATP, the primary energy source in bioenergetics, on demand. Consequently, creatine kinase plays a significant role in energy homeostasis of cells with intermittently high energy requirements. The enzyme is of clinical importance and its levels are routinely used as an indicator of myocardial and skeletal muscle disorders and for the diagnosis of acute myocardial infarction. First identified in 1928, the enzyme has undergone intensive investigation for over 75 years. There are four major isozymes, two cytosolic and two mitochondrial, which form dimers and octamers, respectively. Depending on the pH, the enzyme operates by a random or an ordered bimolecular mechanism, with the equilibrium lying towards phosphocreatine production. Evidence suggests that conversion of creatine to phosphocreatine occurs via the in-line transfer of a phosphoryl group from ATP. A recent X-ray structure of creatine kinase bound to a transition state analog complex confirmed many of the predictions based on kinetic, spectroscopic, and mutagenesis studies. This review summarizes and correlates the more significant mechanistic and structural studies on creatine kinase. PMID- 15804624 TI - RNA-binding proteins in early development. AB - RNA-binding proteins play a major part in the control of gene expression during early development. At this stage, the majority of regulation occurs at the levels of translation and RNA localization. These processes are, in general, mediated by RNA-binding proteins interacting with specific sequence motifs in the 3' untranslated regions of their target RNAs. Although initial work concentrated on the analysis of these sequences and their trans-acting factors, we are now beginning to gain an understanding of the mechanisms by which some of these proteins function. In this review, we will describe a number of different families of RNA-binding proteins, grouping them together on the basis of common regulatory strategies, and emphasizing the recurrent themes that occur, both across different species and as a response to different biological problems. PMID- 15804625 TI - Care homes and dementia. PMID- 15804626 TI - Behavioural symptoms of dementia in residential settings: a selective review of non-pharmacological interventions. AB - For people with dementia living in residential settings, behaviours such as aggression, screaming, restlessness, agitation and wandering are a frequent reason for referral to specialist mental health services for older people. Psychosocial models of dementia have grown in prominence and non-pharmacological interventions have been recommended in professional and government policy statements, either as a first line of treatment or alongside medication. Studies of their effectiveness have been criticised for being poorly controlled, focusing on milder behaviour problems and for requiring a disproportionate use of resources. The recent ruling that risperidone and olanzepine should not be used to control behavioural symptoms in dementia makes it timely to review the evidence for alternative treatments. The current review is a selective one of different types of studies including studies of staff training and liaison interventions, studies of a range of different therapeutic interventions and individualized interventions within a single-case methodology. It is argued that different types of research methodology are appropriate for different studies and that there is still too little evidence to provide firm guidelines. In conclusion, a structured decision-making process for selection of interventions is proposed, in which the limited available evidence can be drawn together to provide a basis for targeting clinical resources while the research evidence is strengthened. PMID- 15804627 TI - Family involvement in residential long-term care: a synthesis and critical review. AB - The objective of this review is to critically synthesize the existing literature on family involvement in residential long-term care. Studies that examined family involvement in various long-term care venues were identified through extensive searches of the literature. Future research and practice must consider the complexity of family structure, adopt longitudinal designs, provide direct empirical links between family involvement and resident outcomes, and offer rigorous evaluation of interventions in order to refine the literature. PMID- 15804628 TI - A qualitative study of the experiences of long-term care for residents with dementia, their relatives and staff. AB - Most older people living in 24-hour care settings have dementia. We employed qualitative interviews to explore positive and negative aspects of the experience of family carers, staff and people with dementia living in 10 homes in London and West Essex, selected to cover the full range of 24-hour long-term care settings. The interview used open semi-structured questions. We interviewed 21 residents, 17 relatives and 30 staff and five main themes were identified: Privacy and choice; relationships (abuse and vulnerability); activities; physical environment; and expectations of a care environment by carers, should they one day live in long-term care themselves. Despite being no longer responsible for the day-to-day care of the residents there was a continuing level of psychological distress among some relatives. We found that residents with a range of severity of dementia were able to participate. The most striking theme from their interviews was the need for choice. All groups talked about improving lines of communication amongst residents, relatives and staff and about the importance of activities. We recommend that homes should set up formal structures for engaging with user and carer views at all levels. This would mean relatives on the board, and regular meetings for residents, relatives, advocates and staff. This should lead to cultural changes where residents are perceived as individuals and care is provided in a more flexible way. There should be a programme of activities in each 24-hour care setting, which all care staff are given time to implement. These activities need to be tailored to the individual resident rather than the whole group. PMID- 15804629 TI - Positive affect among nursing home residents with Alzheimer's dementia: the effect of recreational activity. AB - The experience of positive emotions is an integral component of quality of life. Research suggests that cognitive deficits in persons with dementia may impede their ability to generate pleasurable moments and hence decrease their positive affect. Therefore, structured recreation activities may have the potential to significantly improve resident affect. However, differences in affect between ordinary time and recreation time are not well known. The present study used previously published structured-observation instruments to measure affect and behaviour among 35 dementia residents at two nursing homes in Japan during ordinary time and during recreation time. A total of 3,854 one-minute observations were coded. Dementia residents expressed happiness over seven times more often during recreation time than during ordinary time. Over 60% of ordinary time was solitary, with 65.72% of all observed affect being 'Null Affect'. A total of 43.75% of residents expressed happiness only during recreation time. In addition 48.9% of all behaviour during 'Ordinary Time' was coded as 'Null Behaviour', which indicated that the resident was sitting and doing nothing. Findings indicate that recreation time is significantly higher in positive affect than ordinary time and that virtually all residents benefited from recreation. PMID- 15804630 TI - Down and drowsy? Do apathetic nursing home residents experience low quality of life? AB - This cross-sectional study investigated the relationship between apathy and quality of life (QOL) in nursing home residents (n = 227). In all, 92 residents could be assessed with the Mini Mental State Examination (MMSE), the Geriatric Depression Scale (GDS) and the Philadelphia Geriatric Centre Morale Scale (PGCMS), and were able to answer a question about overall subjective QOL. Apathetic behaviour and consciousness disorders were measured with the Behaviour Rating Scale for Psychogeriatric Inpatients (GIP). Linear regression analysis was first applied to study the association of cognition, depression and consciousness with apathy. It was then used to study the relationship between apathy and QOL, controlling for the constructs that were associated with apathy. The relationship between apathy and QOL appeared to vary with the cognitive functioning of the residents: In residents with a low level of cognitive functioning, apathetic behaviour was associated with high QOL; in residents with a higher level of cognitive functioning, apathetic behaviour was associated with low QOL. The necessity and nature of interventions aimed at stimulating apathetic residents may depend on the level of cognitive functioning of the residents. Further research is needed to determine if and when apathy interventions are appropriate. PMID- 15804631 TI - Mental health problems in the elderly in residential care in Greece--A pilot study. AB - The aim of this study is to examine the mental health problems of older adults living in a residential home in a Greek rural area. A sample of 40 residents was compared with 40 matched controls attending a community open care centre for the elderly (OCCE). The following measures were used: Mini International Neuropsychiatric Interview (MINI), Geriatric Depression Screening Scale (GDSS), and the Mini Mental State Examination (MMSE). The group of the residents had a lower educational level and presented with a higher prevalence of depression and suicidality. Suicidal ideation was not significantly correlated to any of the examined independent social or psychological factors and was persistent during a period of two years follow-up. It is possible that, especially in rural areas, admission in the institution is per se a traumatic event precipitating suicidality. The issues of effective psychological care for older adults in residential care are discussed. PMID- 15804632 TI - The vascular depression hypothesis: the influence of age on the relationship between cerebrovascular risk factors and depressive symptoms in community dwelling elders. AB - The current study examined the moderating effect of age on the relationship between cerebrovascular risk factors (CVRF's) and depressive symptoms. Consistent with the broader vascular depression model, it was hypothesized that CVRF's would demonstrate a stronger link to depressive symptoms in the older age groups than among the younger age groups. Data from 2916 adults from the Resources, Stress, and Older Persons Panel Study were utilized. Path analysis was used to estimate direct and indirect effects (via health related symptoms and limitations) of CVRF's on depressive symptoms. Path analyses were estimated separately on four age groups: 50-64 years old, 65-74 years old, 75-84 years old, and 85 years and older. CVRF's and other comorbid medical conditions were highly predictive of health related symptoms and limitations across the four age groups. Health related symptoms and limitations were strongly linked to depressive symptoms and mediated the influence of medical illnesses (both vascular and nonvascular) on depressive symptoms. However, CVRF's exerted a unique effect on depressive symptoms in the oldest-old group (i.e., 85+). Among those over the age of 85, a greater number of CVRF's was associated with more severe depressive symptoms independent of health related symptoms/limitations and other comorbid medical conditions. Health related symptoms and limitations mediated the relationship between CVRF's and depression in individuals under 85. That is, the influence of vascular burden on depression is predominately indirect via health related limitations. But among those over the age of 85, vascular disease had a unique contribution on depression, even after controlling for other comorbid medical illness and health related limitations. This finding supports the vascular depression hypothesis and is consistent with prior work suggesting vascular disease may exert its greatest effect on depression in the context of increasing frailty. PMID- 15804633 TI - Attachment behaviours and parent fixation in people with dementia: the role of cognitive functioning and pre-morbid attachment style. AB - This study replicates and extends exploratory research into the occurrence of attachment behaviours and parent fixation amongst people with dementia. Relationships between cognitive functioning, pre-morbid attachment style, attachment behaviours and parent fixation were examined. Fifty-three people with dementia, living in residential or nursing homes, completed the Standardised Mini Mental State Examination and were interviewed about their parents. A family member or friend rated pre-morbid attachment style and care staff made observations of attachment behaviour. Results indicated that parent fixation occurred more often in participants with lower levels of cognitive functioning. Parent fixation was not related to pre-morbid attachment style. The occurrence of overt attachment behaviour was inconsistently associated with both high and low levels of cognitive functioning, at different times of the day. Participants with an avoidant attachment style exhibited more overt attachment behaviour than participants with a secure attachment style. Findings are interpreted in terms of attachment theory and the clinical and research implications of the study are discussed. PMID- 15804634 TI - The psychological well-being of the Chinese elderly living in old urban areas of Hong Kong: a social perspective. AB - Using the Geriatric Depression Scale-15 (GDS-15) and the short form of the General Health Questionnaire-12 (GHQ-12), this study found that the Chinese elderly who live in old urban areas of Hong Kong are at risk of suffering from poor mental health because of their relatively deprived social conditions. It was also found that the respondent's scores in the GDS-15 correlated with their social conditions, including type of housing, availability of helpers, financial status and family support. However, Chinese elderly people often avoid seeking help because of the fear of being stigmatised. There is a need for more research in the future to study their needs and problems so that social work practitioners can be well informed and culturally competent to help this disadvantaged group. PMID- 15804635 TI - Assessing Chinese older adults' suicidal ideation: Chinese version of the Geriatric Suicide Ideation Scale. AB - The psychometric properties of the Chinese version of the Geriatric Suicide Ideation Scale (GSIS-C) were investigated in a sample of 154 Hong Kong Chinese older adults. The internal consistency, test-retest reliability, face validity, convergent and divergent validities of the GSIS-C were assessed and all of them were well above acceptable levels. The GSIS-C demonstrated excellent internal consistency including the four subscales and the test-retest reliability of the GSIS-C was also good. In terms of convergent validity, the GSIS-C correlated significantly and positively with depression (assessed by CES-D), loneliness (assessed by Revised UCLA Loneliness Scale), and hopelessness (assessed by Beck 'Hop'elessness Scale). The divergent validity of the GSIS-C was demonstrated by the negative but significant, association between the GSIS-C and two variables including self-rated health status and life satisfaction (assessed by Life Satisfaction Inventory--Version A). PMID- 15804636 TI - Utilization of emergency center services by older adults with a psychiatric diagnosis. AB - Information regarding older adult emergency center (EC) patient characteristics remains limited, despite its increasing importance in health care delivery systems. This retrospective study encompasses all EC visits over an eight-year period (n = 825,682) to a large urban county hospital. Only participants with a primary psychiatric diagnosis were examined, and included a total of 53,894 adults, 18-64 years old and 1,478 adults, > or =65 years old. Despite an increasing aging population, EC visits for older adults with psychiatric disorders did not increase over time. Within the older adult sample, cognitive, psychotic, and bipolar disorders were associated with higher rates of admission to the hospital, while substance use, depressive, and anxiety disorders were associated with lower numbers of inpatient admissions. African-Americans were over-represented in the EC and admitted to the hospital at higher rates, compared to other ethnic groups. Caucasian patients were the group most frequently diagnosed with a substance use disorder. In conclusion, differences in race, and diagnosis support the idea that such variables directly relate to utilization rates, presentation, and disposition within the EC. PMID- 15804637 TI - Everyday competence and depressive symptoms: social support and sense of control as mediators or moderators? AB - Depression is common among the elderly members of Hong Kong Chinese society. The objectives of the present study are to assess the relationship between everyday competence and depressive symptoms and to test whether sense of control and social support mediate and moderate the impact of deterioration in everyday competence on depressive symptoms. The respondents were 393 people aged 60 years and older from a cross-sectional study of a representative community sample of the elderly population in Hong Kong and they were interviewed using a structured questionnaire. In multiple regression analyses, we found that everyday competence was significantly and negatively related to depressive symptoms (beta = -0.26, p < 0.01) after we had adjusted age, gender, marital status, years of education, self-rated health status, and number of chronic illnesses. Moreover, both sense of control and social support were mediators in the linkage between everyday competence and depression. However, neither sense of control nor social support moderated the effect of everyday competence on depression. Findings suggest that both sense of control and social support play important roles in the relationship between everyday competence and depression. PMID- 15804640 TI - Coping with conflict. PMID- 15804641 TI - Psychomotor skills for the undergraduate medical curriculum in a developing country--Pakistan. AB - OBJECTIVE: To identify essential psychomotor skills for all the medical graduates of an undergraduate programme in Pakistan. MATERIALS AND METHODS: Twenty-five physicians practising in a tertiary care centre and ninety primary care physicians used a Likert's scale, ranging from "very essential" to"not required at all", to mark 99 psychomotor skills in the undergraduate medical curriculum in Pakistan. RESULTS: Overall the opinions of both the groups about the essential skills matched except for a few areas. CONCLUSION: This study provides baseline data about psychomotor skills that a medical graduate in developing countries should be able to perform. Further studies will be undertaken by involving other stakeholders to identify and incorporate these skills in the undergraduate medical curriculum, thereby enabling graduates to practice in all the settings in Pakistan. PMID- 15804642 TI - Assumptions about disease treatment challenged in a family health clerkship: views of first clinical year medical students. AB - CONTEXT: During a family health clerkship at the University of The West Indies, students are expected to acquire individual and community diagnosis skills and the ability to relate the two, as well as acquire knowledge of other community agencies involved in health care. OBJECTIVE: To determine the main assumptions related to disease treatment, which students have had to re-think after engaging in this clerkship. METHODS: End of clerkship assessments were carried out from two successive groups of third year medical students (n=64) at the University of West Indies, Jamaica. Students were asked to "list two assumptions regarding treating disease that have been challenged by your experience". A subsequent content analysis was done. RESULTS: Fifty-five students (86%) completed the assessment. All assumptions were listed (n=99) and similar issues were linked into emerging themes. Twenty-five groups of assumptions produced seven main themes: "Issues related to compliance" (27.3%),"Patient's treatment is mainly physical" (17.2%), " Superiority of western medicine over alternative" (15.2%), "Patients' health seeking behavior and attitudes" (12.1%), " The extent of the contribution of social factors on health" (12.1%) and "Patients' knowledge and understanding of health" (7.0%). CONCLUSION: The majority of students examined felt challenged on three themes: relating to issues of compliance, treating the "whole" patient not just the physical, and the superiority of western medicine over alternative. The three most popular individual assumptions were; patients have a mindset that favours compliance, medication affordability does not affect compliance and treatment is independent of social and environmental conditions. PMID- 15804643 TI - From systems to tissues: a revolution in learning in perioperative education. AB - AIM: This paper presents the results of an exploratory project undertaken to evaluate the change to teaching and learning of surgical specialties in the Postgraduate Diploma in Advanced Nursing (Perioperative) conducted at La Trobe University,Victoria, Australia. BACKGROUND: The teaching of perioperative nursing practices has traditionally been based on the medical model of surgical specializations, using information transmission as the main teaching approach. It has become clear to perioperative educators that it is possible to look at the basic theoretical principles involved in the teaching of surgical interventions from a different perspective. METHOD: This project is an exploratory descriptive study that employed a qualitative approach, using a reflective approach to teaching, results of a focus group discussion and subject assessment as data. Research Ethics Committee protocols were followed. Aversion of nominal group technique was used for the focus group interview. OUTCOMES: The restructure of this subject shows potential in bringing about a conceptual shift in nurse education in three areas: (1) Organizational principle or curriculum structure: from content focus to theoretical principles; (2) Learning mode: student involvement from passive student to professional decision maker; (3) Teaching strategy:from teacher-focused strategy with intentions to transmit information to a student-focused strategy aimed at students changing their conceptions. CONCLUSION: This study suggests that perioperative students are able to demonstrate critical thinking and apply principles of surgical technique across a range of specialty areas without having participated in the particular surgery beforehand. PMID- 15804644 TI - Influence of an interprofessional HIV/AIDS education program on role perception, attitudes and teamwork skills of undergraduate health sciences students. AB - PURPOSE: An evaluation study of an undergraduate HIV/AIDS interprofessional education program for medical, nursing and pharmacy students was undertaken to assess changes in role perception, attitudes towards collaboration, self-reported teamwork skills and satisfaction with a shared learning experience. METHODS: A combined one group pretest-posttest and time-series study design was used. Several survey instruments and observation checklists were completed by students and tutors before, during and after the educational program. RESULTS: Students reported greater awareness of roles and the continuous exposure to interprofessional learning led to improved attitudes towards teamwork. Standardized patients were effective in fostering an experience of realism and motivating collaboration between students. CONCLUSION: A problem-based learning approach combined with standardized patients was effective in enhancing HIV/AIDS interprofessional role perception, enhancing attitudes towards collaboration and interprofessional approaches to HIV/AIDS care and fostering confidence in teamwork skills among pre-licensure health sciences students. PMID- 15804645 TI - On the additional value of lectures in a problem-based curriculum. AB - OBJECTIVE: The purpose of this article is to report on a study conducted to investigate the additional value of lectures in problem-based learning. We hypothesized that lecture quality, as indicated by students, would have a positive influence on time students would spend on self-study, that they would increase intrinsic interest in subject matter and would have a positive influence on achievement. Lecture quality would, in this view, add to other factors that play a role in problem-based learning, such as prior knowledge of students, the quality of problems presented, the functioning of the tutor and small-group collaboration. METHOD: The hypothesis on the influence of lecture quality on learning was assessed by means of testing a structural equating model, using data from 1500 students. DISCUSSION AND CONCLUSIONS: We concluded that lecture quality does not affect time spent on study, does not add to achievement and does not influence intrinsic interest in subject matter. Post-hoc analysis, however, demonstrated the quality of lectures was related to other components of problem based learning, not part of the presented model, such as students' perceptions of lectures as organizer of their study efforts or lectures as a means of putting the problems into a broader perspective. PMID- 15804646 TI - Revitalising PBL groups: evaluating PBL with study teams. AB - CONTEXT: In problem-based learning (PBL), students are actively engaged with psychological learning principles as activation of prior knowledge, elaboration and organization of knowledge. In their tutorial groups, however, students do not always apply these principles when working with a procedure like the "Seven-Jump" method. To stimulate students to use these principles more often, they were offered another format within a PBL context: PBL with study teams. During the period of self-study, students work on a regular basis in so-called study teams, small groups of 3-4 persons. In these groups they explain to each other their learning outcomes, clarify for each other their problems while studying texts and organize their knowledge to present this to the members of other study teams in their tutorial group. Previous research showed that students spent more time on self-study in a PBL with study team condition than in a traditional PBL context. In this study the achievement as well as appreciation of students participating in a PBL with study teams' environment, is compared with students working in a traditional PBL environment. OBJECTIVES: To determine whether PBL with study teams differs from the traditional PBL environment in students' appreciation and study time. METHODS: We conducted an experiment in two blocks over two years. Questionnaires were administered to collect data on appreciation and time for self-study. RESULTS: Students' appreciation of the two formats did not differ much. The large standard deviations indicate considerable differences in appreciation between individual students. Appreciation was slightly higher in the second experiment when instructions about how to collaborate were less strict. Students devoted twice as many hours studying in the study group format compared with the traditional PBL format. CONCLUSIONS: The students indicated that they enjoyed the format but that the increased workload disturbed their customary study rhythm. Assessment scores and tutors'impressions suggest that study teams foster deeper learning for understanding. Whether this can be attributed to more intensive individual and collaborative elaboration activities should be the subject of further studies. PMID- 15804647 TI - Advice to deans: assessing faculty work by "sitting beside". PMID- 15804648 TI - Using behavior change theories to enhance hand hygiene behavior. PMID- 15804649 TI - Service-learning risk management considerations: the community visitor project. PMID- 15804650 TI - An interview of Lizo Mazwai. Interview by Jane Westberg. PMID- 15804651 TI - An interview of Mary MacLean. Interview by Jane Westberg. PMID- 15804652 TI - An interview of Erik Scherman. Interview by Jan van Dalen. PMID- 15804658 TI - 'We are not just participants--we are in charge': the NACCHO ear trial and the process for Aboriginal community-controlled health research. AB - OBJECTIVE: Methodological criteria that characterise ethically sound community based studies are often described in overviews but are rarely documented in clinical studies. Research investigating the health of Aboriginal Australians is often small-scale, descriptive and largely driven by non-Indigenous people. The 'community-controlled' model of research relating to Aboriginal peoples health is a form of 'participatory' research that shifts the balance of control towards those being researched. This paper describes the methodological issues and principles that underpin community-controlled health research; their practical application; and encourages their adoption in research involving Indigenous populations. DESIGN: Descriptive report of the methods used to conduct the landmark Aboriginal community-controlled multi-centre double-blind randomised controlled clinical ear trial investigating ototopical treatments for chronic suppurative otitis media. RESULTS: The characteristics of the community controlled research model are illustrated under the headings of: setting the research agenda; research project planning and approval; conduct of research; and analysis, dissemination and application of findings. CONCLUSION: The 22 methodological elements which defined the community-controlled design of the ear trial may assist community groups, external research bodies and funding agencies to improve the acceptability, quality and scope of research involving Indigenous peoples. Aboriginal community-controlled organisations are well placed to lead research, which can be interventional and of a high scientific standard without compromising the values and principles of those being researched. With over 120 Aboriginal community-controlled health services (ACCHSs) across Australia, the potential exists for these services to engage in multi-centre research to realise solutions to health problems faced by Indigenous Australians. PMID- 15804659 TI - Ethnicity, socio-economic status, overweight and underweight in East London adolescents. AB - BACKGROUND: The developed world is experiencing an 'epidemic' of childhood obesity but little is known about the prevalence of obesity, or underweight, amongst adolescents from minority ethnic groups in the UK. An understanding of the prevalence of obesity and overweight in these populations is important since some ethnic groups may be particularly vulnerable to the adverse health effects associated with obesity. STUDY OBJECTIVES: To examine levels of extreme obesity, obesity, overweight and underweight amongst a representative sample of adolescents from different ethnic groups in East London and to explore the association between socio-economic status and body mass index (BMI). DESIGN: A school-based survey of adolescents aged 11-14. Obesity and overweight were estimated using the 1990 UK growth reference (UK 90) and the International Obesity Task Force (IOTF) cut-off points. Extreme obesity was defined as a BMI more than three SD above the UK 90 mean. Underweight was examined by looking at those with a BMI below the 15th or the 5th UK 90 percentiles. MAIN RESULTS: A total of 2,482 adolescents were surveyed (response rate 84%), 73% from non-white ethnic groups. Although there were significant differences in BMI between ethnic groups, high levels of overweight were seen in all ethnic groups. More than one third were overweight and one-fifth were obese using the UK 90; and over a quarter were overweight and almost one-tenth were obese using the IOTF cut-offs. Two per cent were extremely obese. Indian males were at higher risk of being overweight than white British males. The prevalence of obesity and overweight was similar in white British and Bangladeshi males. Overall the prevalence of underweight was slightly lower than that predicted by the UK 90, but South Asian ethnic groups, especially males, had a higher prevalence of underweight than other groups. No associations between BMI and measures of socio-economic status were found in this relatively deprived population. CONCLUSIONS: The 'epidemic' of childhood obesity observed in the UK involves adolescents from all ethnic groups, although there are some differences between ethnic groups in the prevalence of overweight. Indian males appear to be at increased risk of being overweight. There is no evidence of a simultaneous increase in underweight amongst this population overall, but Bangladeshi, Indian and Pakistani boys appear to be at increased risk of being underweight. PMID- 15804660 TI - Unaccompanied young adult refugees in Sweden, experiences of their life situation and well-being: a qualitative follow-up study. AB - OBJECTIVE: In the late 1980s unaccompanied children began arriving in Sweden. Many of them were teenage boys who had been called up or were at risk of being called up for military service in a war in their home country. Others had fled their homeland for political reasons. The purpose of the study was to describe how unaccompanied young adult refugees experienced their own life situation and well-being seven years after they had gained permanent residency. DESIGN: The original sample comprised 34 unaccompanied refugees who at 16-26 years of age were placed in a municipality in Sweden. Eleven of these participated in the present second follow-up study after a mean of 10 years in their new country. Qualitative interviews were conducted and analysed according to Giorgi's descriptive phenomenology. RESULTS: Most of the participants expressed contentedness with their lives and had begun to adjust to their new country. They had a social network of friends from the same ethnic group, and their Swedish contacts were mostly workmates. A few felt lonely and expressed despondency and depression. They were single and reported a small network and limited social support. One participant described symptoms of post-traumatic stress disorder (PTSD), even after 11 years in the new homeland. CONCLUSION: It appears that most of the participants in the present follow-up study had worked through the problems that typically face refugees and had begun adapting to their new country, while a few still felt lonely and did not feel part of the community. The results may be affected by non-response. Those who dropped out may live with more distress in everyday life and suffer depression more frequently. PMID- 15804661 TI - Psychosocial Experiences of East and Southeast Asian men who use gay Internet chatrooms in Toronto: an implication for HIV/AIDS prevention. AB - OBJECTIVES: In recent years we have witnessed an increase in Asian men who use gay Internet chatrooms in Toronto. Previous research has shown that many men who had sex with men (MSM) sought sex partners through the Internet and that meeting sex partners via the Internet increases sexually transmitted infection (STI) and HIV risk. This study aims to (1) explore psychosocial issues relating to Asian men who use gay chatrooms and (2) identify culturally appropriate HIV prevention strategies for this population. DESIGN: In-depth interviews were conducted with a total of 21 East and Southeast Asian men who used Internet gay chatrooms. Unstructured, open-ended questions were used to obtain narrative data to help understand their lived, psychosocial experiences of gay chatrooms. Transcripts of the interviews were read to highlight themes and concepts. RESULTS: Analysis revealed complex lived, psychosocial experiences of Asian men who use gay chatrooms in Toronto. They tended to be socially isolated and highly marginalized, which had led to intense needs for social connections and thus left some Asian men vulnerable to sexual exploitation. Although they were fully aware that they should use condoms in anal intercourse with a casual partner, they had some misconceptions about HIV. Moreover, they rarely, if ever, used condoms in oral sex with a casual partner, which might leave them vulnerable to STI. CONCLUSIONS: It is important for service providers to continually provide accurate information about STIs and HIV/AIDS including how they can be contracted. However, HIV prevention strategies for this population must also address issues relating to social isolation and marginalization in order to combat the spread of HIV/AIDS effectively. This can be accomplished by an online peer support program. PMID- 15804662 TI - Symptoms of distress and posttraumatic stress among South African former political detainees. AB - Considerable debate has centred on the question of traumatisation among individuals who have survived human rights violations in societies that have undergone political conflict. In order to gain an estimate of the extent of long term traumatisation among political activists who experienced torture and abuse in detention during the apartheid era in South Africa, a sample of 148 survivors of such experiences were recruited in a cross-sectional study and asked to complete the Hopkins Symptom Checklist (HSCL), the Impact of Event Scale (IES), and the Trauma Symptoms section of the Harvard Trauma Questionnaire (HTQ). The proportions of the sample that scored above the clinical cut-points on these measures were calculated. On the HSCL, 14.19% of the sample scored above the cut point for clinical significance of 44; on the IES, 17.57% scored above the clinical cut-point of 44; and on the HTQ, 37.83% scored above the cut-point of 75. Moreover, the sample's mean scores were significantly higher than the cut point for clinically significant distress on the HSCL (p < 0.001); significantly lower than the cut-point for severe traumatisation on the IES (p < 0.001); and non-significantly lower than the cut-point for clinically significant traumatisation on the HTQ (p = 0.074). These results are considered in terms of current theoretical debates on the relevance and applicability of posttraumatic stress disorder as a circumscribed nosological entity in developing countries that are in the process of coming to terms with a history of political conflict. PMID- 15804663 TI - Childhood diarrhoea in Ankara, Turkey: epidemiological and clinical features of rotavirus-positive versus rotavirus-negative cases. AB - Published reports dealing with rotavirus infections in Turkey are very scarce. This study included 1099 consecutive paediatric patients with diarrhoea, who sought care at 3 hospitals in Ankara, Turkey between 1999 and 2002 and were investigated for the presence of rotavirus antigen in faeces. Rotavirus antigen was detected by an immunochromatographic test, Simple Rotavirus (Operon, Spain). Other clinical and laboratory data were extracted from patient journals. A total of 404 (36.8%) patients were positive for rotavirus antigen. Rotavirus antigen was more frequently detected in boys than girls (40.8 vs 31.8%) and in children younger than 2 y (62.7%). The proportion of rotavirus-positive children was higher in the winter season (49.6%; November to April) and the highest proportion was observed in December (55.4%). Rotavirus-associated diarrhoea had a more severe clinical presentation than non-rotaviral diarrhoea; 55.3% of all patients who required hospitalization were rotavirus-positive. The seasonal and epidemiological characteristics of rotavirus diarrhoea in Ankara were similar to those in the USA and Europe. For reliable nationwide information about the epidemiology of rotavirus-associated disease in Turkey, more individual studies and reliable official statistics of gastroenteritis cases are needed. PMID- 15804664 TI - Distribution of gentamicin from a Gentacoll sponge measured by in vivo microdialysis. AB - Microdialysis has produced valuable information concerning the distribution of antibiotics in peripheral tissue. The aim of this study was to measure the local distribution of gentamicin in cancellous bone tissue after the application of a Gentacoll sponge. Two microdialysis catheters were inserted into the cancellous bone of 9 pigs. After calibration of the microdialysis, a Gentacoll sponge was implanted, either dry or wet, into the bone marrow of the tibia. Serum and microdialysates were obtained over a period of 6 h. Data presented are median (range). A rank sum test was performed for statistical analysis. The peak concentration was 120 mg/l (33-585) (wet group) and 178 mg/l (60-1294) (dry group) (p=0.31). The AUC6 of the catheters placed 1 cm away from the implant was 24,431 mg*min/l (5155-152,855) and the AUC6 of the catheter placed 2 cm away from the implant was 13,759 mg*min/l (6351-74,573) (p=0.25). The study showed that peak concentrations and AUC6 did not differ between the wet- and dry-application group; neither did the distance from the sponge to the catheter have a significant impact on the distribution of gentamicin. It seems that microdialysis is capable to measure concentrations of locally applied antibiotics. PMID- 15804665 TI - VTEC O117:K1:H7. A new clonal group of E. coli associated with persistent diarrhoea in Danish travellers. AB - 20 patients were infected with a new group of verocytotoxin-producing E. coli (VTEC) strains of serotype O117:K1:H7 during a 5-y period. The main symptoms were persistent watery diarrhoea with abdominal cramps; 2 persons of the 20 were healthy carriers. The duration of gastrointestinal symptoms in patients was median 11 weeks with 80% being ill for more than 30 d. In 19 cases the infection was acquired during travel (Asia, Africa and Cuba), and 1 case was laboratory acquired. All strains were positive for the vtx1 gene and negative for the vtx2, the eae, the saa and the ehxA genes. 13 strains (65%) were resistant to 4 or more antimicrobial agents. By PFGE using the restriction enzyme XbaI, the strains were clonally related, but not identical. O117:K1:H7 is a clonal group of VTEC that should be considered in patients returning from Africa and Asia with long-lasting watery diarrhoea. PMID- 15804666 TI - Accidental exposure to biological material in healthcare workers at a university hospital: Evaluation and follow-up of 404 cases. AB - The care and follow-up provided to healthcare workers (HCWs) from a large teaching hospital who were exposed to biological material between 1 August 1998 and 31 January 2002 is described here. After exposure, the HCW is evaluated by a nurse and doctor in an emergency consultation and receives follow-up counselling. The collection of 10 ml of blood sample from each HCW and its source patient, when known, is made for immunoenzymatic testing for HIV, HBV and HCV. Evaluation and follow-up of 404 cases revealed that the exposures were concentrated in only a few areas of the hospital; 83% of the HCWs exposed were seen by a doctor responsible for the prophylaxis up to 3 h after exposure. Blood was involved in 76.7% (309) of the exposures. The patient source of the biological material was known in 80.7% (326) of the exposed individuals studied; 80 (24.5%) sources had serological evidence of infection with 1 or more agents: 16.2% were anti-HCV positive, 3.8% were HAgBs positive and 10.9% were anti-HIV positive. 67% (273) of the study population completed the proposed follow-up. No confirmed seroconversion occurred. In conclusion, the observed adherence to the follow-up was quite low, and measures to improve it must be taken. Surprisingly, no difference in adherence to the follow-up was observed among those exposed HCW at risk, i.e. those with an infected or unknown source patient. Analysis of post exposure management revealed excess prescription of antiretroviral drugs, vaccine and immunoglobulin. Infection by HCV is the most important risk of concern, in our hospital, in accidents with biological material. PMID- 15804667 TI - Septic arthritis due to Salmonella enteritidis associated with infliximab use. AB - A unique case of septic arthritis caused by Salmonella enteritidis in a patient receiving infliximab for rheumatoid arthritis is presented. Antimicrobial chemotherapy with surgical intervention was necessary for eradication of the infection. Physicians should be aware of rare manifestations of Salmonella infections associated with infliximab use, especially in endemic areas. PMID- 15804668 TI - Drotrecogin alpha (activated) in neonatal septic shock. AB - A 12-d-old neonate suffering from group B streptococcal septic shock was treated with 24 microg/kg/h recombinant human activated protein C [rhAPC, drotrecogin alpha (activated)] for 96 h. The protein C activity increased from 5% to 53% after rhAPC infusion. The patient recovered within 14 d without any adverse effects. PMID- 15804669 TI - Haemorrhagic cellulitis caused by Salmonella enteritidis. AB - Soft tissue infections are rare manifestations of extra-intestinal salmonellosis and occur more frequently in immunocompromized patients [1-4]. We report haemorrhagic cellulitis in an immunosuppressed patient with sepsis caused by a non-typhoid Salmonella. PMID- 15804670 TI - Candida albicans tricuspid valve endocarditis in an intravenous drug addict: successful treatment with fluconazole. AB - An addicted, intravenous drug user was treated for Candida albicans tricuspid valve endocarditis with high dose fluconazole for 8 months, without relapse after 30 months. PMID- 15804671 TI - Bio-hazards and drug reactions: a cautionary tale. AB - Laboratory acquired infection with toxoplasmosis has been described. The clinical features in this situation can vary from asymptomatic to severe disease. Prophylactic therapy is recommended on the basis of clinical experience and case reports. We describe a case in a young man, where prophylaxis resulted in neutropenia complicated by subsequent Listeria meningitis. PMID- 15804672 TI - Septic right superior ophthalmic vein thrombosis complicated by left sphenoid sinusitis. AB - We report a case of septic right superior ophthalmic vein thrombosis complicated by left sphenoid sinusitis with methicillin-sensitive Staphylococcus aureus (MSSA). Early recognition by clinical symptoms, and computed tonography (CT) and magnetic resolution image (MRI) of the head and sinus is needed for prompt surgical drainage of the sinus and appropriate antimicrobial therapy. The use of steroids and anticoagulation for this condition is still debated. PMID- 15804674 TI - A sensitization-homeostasis model of nicotine craving, withdrawal, and tolerance: integrating the clinical and basic science literature. AB - Recent reports suggest that nicotine withdrawal symptoms are common among adolescents after a few weeks of intermittent tobacco use. No current model of nicotine dependence had predicted the rapid development of symptoms of dependence and withdrawal before the development of tolerance. We present a model that integrates neuroscience with clinical observations regarding how nicotine dependence develops, progresses, and resolves in humans. The central tenet of this sensitization-homeostasis model is that nicotine's dependence liability derives from its ability to stimulate neural pathways responsible for the suppression of craving. As a result of sensitization, the craving suppression produced by nicotine is magnified to superphysiological levels. The overinhibition of neurons responsible for craving initiates compensatory homeostatic measures that stimulate the craving pathways and result in craving when nicotine is absent. Separate homeostatic mechanisms are responsible for craving, withdrawal, and tolerance. The sensitization-homeostasis model is unique in its attribution of dependence to craving suppression, its attention to the temporal relationships among clinical features of nicotine dependence, and its extensive integration of clinical observations and basic science. It provides a framework for theory-based research. PMID- 15804675 TI - Playing hide-and-seek with the tobacco industry. AB - Despite many peer-reviewed works that draw on tobacco industry documents that have now been made public, questions remain about how complete a picture has emerged. We present a conceptual framework that identifies and evaluates tobacco industry efforts to conceal information. Widespread document destruction like that in recent litigation in Australia is just one of more than a dozen tobacco industry efforts to prevent access, or at least timely access, to documents. Industry efforts range from small, locally employed initiatives to company-wide tactics. Some efforts, such as using "oral only" procedures, scrambling telephone lines, or involving lawyers in scientific projects, are preemptive. Others seek to deal with already existing documents by invoking bogus claims of legal privilege, stipulating "read then destroy" for memos, and rewriting problematic memos. That evidence of concealment has, in fact, been found in tobacco company archives attests to the futility of attempting to control the flow of millions of pieces of paper among tens of thousands of employees. However, researchers have yet to reveal the full story: We know of the industry's failures in concealing information, but not its successes. The industry's objective is not destruction of information per se, but prevention of public disclosure of that information. Exposing the tobacco industry's many approaches to concealment provides greater insight into companies' intentions and potential means for stripping away that concealment. PMID- 15804676 TI - A family smoking index to capture genetic influence in smoking: rationale and two validation studies. AB - Despite a growing appreciation that genetic factors may impart vulnerability toward smoking behavior, only a modest consensus has been created about the specific genetic mechanisms that may underlie various aspects of smoking. A core feature of genetic contribution toward any complex human behavior is familial resemblance. Most previous attempts to index familial smoking have classified individuals into discrete categories, based on the number of smokers in a family. We discuss the development of a continuous measure of familial smoking, the Family Smoking Index (FSI), which is based on the proportion of smokers in first- and second-degree family members and provides a more precise weighting according to genetic proximity. We present the psychometric characteristics of the FSI as well as initial validation data from two studies. We also describe current and future directions for continued FSI validation and application. PMID- 15804677 TI - A self-administered questionnaire to measure cigarette withdrawal symptoms: the Cigarette Withdrawal Scale. AB - Because few tobacco withdrawal scales have been submitted to appropriate validity analyses, we sought to develop and assess the validity of a new, self administered scale measuring cigarette withdrawal symptoms. We generated the instrument content by conducting a qualitative survey of 404 smokers and ex smokers. Then we tested 61 items on the Internet in 3,050 smokers and ex-smokers. Subsamples provided comprehensive retest data after 17 days (n = 1218) and smoking status after 41 days (n = 673). The study resulted in a 21-item, six dimension scale labeled the Cigarette Withdrawal Scale (CWS-21). The six subscales cover the main components of nicotine or tobacco withdrawal in the Diagnostic and Statistical Manual of Mental Disorders and International Statistical Classification of Diseases and Related Health Problems and in qualitative data: Depression-anxiety, craving, irritability-impatience, appetite weight gain, insomnia, and difficulty concentrating. The six scores had a satisfactory test-retest reliability (r = .60-.71) and a high internal consistency (Cronbach's alpha = .83-.96). The factor structure of the scale was robust in a bootstrap resampling procedure. In ex-smokers, all scores except appetite-weight gain and insomnia predicted relapse at 41-day follow-up. In recent ex-smokers who had quit smoking less than 14 days before baseline, all scores except appetite-weight gain decreased between baseline and the 17-day retest. In baseline ex-smokers who relapsed to smoking at the 17-day retest, appetite-weight gain decreased and craving increased between baseline and retest. CWS-21 is a reliable, valid, multidimensional measure of cigarette withdrawal symptoms that is sensitive to change over time and predicts relapse to smoking. PMID- 15804678 TI - Smoking cessation patterns and predictors among adult Californians of Korean descent. AB - This paper describes smoking cessation among adult California residents of Korean descent, based on a telephone survey (N = 2,830). The overall quit rate (number of former smokers divided by number of ever-smokers) was 55.0% (55.8% among males and 49.6% among females). Acculturation was negatively associated with 90-day abstinence after controlling for demographic, social support, health belief, environmental, lifestyle, and respiratory illness history variables (OR = 0.59, 95% CI = 0.38-0.91). Those with lower acculturation, higher body mass index, a social network discouraging smoking, home smoking restriction, correct beliefs about smoking harm, and significant concern about one's health were most likely to quit smoking for 90 days or longer. Results suggest that social and situational conditions should be included in the design of smoking cessation interventions among Koreans. PMID- 15804679 TI - Differences in risk-taking propensity across inner-city adolescent ever- and never-smokers. AB - Because adolescent smoking is a significant public health concern, potential value lies in understanding and identifying the psychological factors that distinguish ever- and never-smokers. To that end, we examined the relationship between risk-taking propensity as measured by the Balloon Analogue Risk Task and ever-smoking (i.e., even one puff) versus never-smoking in a sample of 125 predominantly African American high-school adolescents (M = 15.1, SD = 1.5). Results indicated that ever-smokers and never-smokers differed on risk-taking propensity; further risk-taking propensity was related to smoking status above and beyond both demographic variables and a measure of self-reported impulsive sensation seeking. We discuss these results in relation to the potential utility of a multimethod assessment approach (i.e., self-report measures and behavioral tasks) to identify adolescents' risk-taking susceptibilities and engagement in smoking and other risk-taking behaviors. PMID- 15804680 TI - Development and validation of an adolescent smoking consequences questionnaire. AB - Several researchers have investigated the role of outcome expectancies in the initiation and maintenance of smoking behavior. Empirical studies with adults, using a validated self-report instrument, have shown that smokers identify higher levels of positive smoking expectancies and lower levels of negative smoking expectancies compared with nonsmokers and ex-smokers. Studies examining smoking outcome expectancies among adolescents have yielded similar findings, but many of these studies did not use a self-report expectancy measure validated with teens. Therefore, the present study sought to modify a well-known adult smoking expectancy instrument, the Smoking Consequences Questionnaire (SCQ), and to validate its factor structure with a community sample of adolescent nonsmokers and current smokers (aged 11-19 years). Results of a confirmatory factor analysis provided support for a seven-latent-factor structure modeled after the SCQ-Adult. As expected, smoking behavior and the intent to smoke in the future were associated with the endorsement of positive and negative smoking outcome expectancies. The psychometric data provide early support for the validity and reliability of this modified SCQ, renamed the Adolescent Smoking Consequences Questionnaire, for the assessment of smoking expectancies among smoking inexperienced and experienced teens. PMID- 15804681 TI - Nicotine dependence, depression, and gender: characterizing phenotypes based on withdrawal discomfort, response to smoking, and ability to abstain. AB - Smoking is often viewed as a comprehensive phenotype rather than a complex set of traits involving intermediate phenotypes. To explore this issue in a laboratory setting, we tested 69 smokers stratified on depression, nicotine dependence, and gender. On the third day of an initial withdrawal period, we tested for differences among participants in uncued and cued craving and withdrawal; on the fourth day, we exposed them to a controlled dose of smoke and assessed them for physiological and hedonic effects and reduction of craving and withdrawal. Following resumption of smoking for at least a week, we then tested participants on their ability to abstain for an 11-day interval. During the withdrawal test, high-depressed smokers and men exhibited elevated craving and withdrawal scores overall, whereas no differences emerged for dependence. Cue exposure produced significant increases in craving but not withdrawal. During the smoke-exposure test, men were significantly more likely than women, and high-depressed smokers more likely than low-depressed smokers, to show evidence of experiencing pleasurable "buzzes." High-dependent smokers showed significant increases in diastolic blood pressure, possibly suggestive of greater sensitivity to nicotine. During the quit test, high-dependent smokers had more difficulty abstaining than low-dependent smokers, and women more than men; no differences emerged based on depression. Independently of group membership, inability to abstain was predicted by increased anxiety, depression, and difficulty concentrating in response to cue exposure. These findings provide support for the existence of phenotypes that can be distinguished by withdrawal symptomatology (primarily driven by depression) and ability to remain abstinent (primarily driven by dependence). PMID- 15804682 TI - Prevalence and predictors of smoking behavior among Vietnamese men living in California. AB - Vietnamese populations in Vietnam and the United States have a high prevalence of smoking. The associations among behavioral risk factors, acculturation, and smoking among the Vietnamese population living in the United States are not well documented. The present study aimed to identify the factors associated with smoking behavior among Vietnamese men living in Santa Clara County, California. A cross-sectional random-digit-dialed telephone survey was conducted. The sampling frame consisted of 27 Vietnamese surnames from the Santa Clara County telephone directory. A total of 660 adult respondents were interviewed to collect information on general health status, alcohol and tobacco use, HIV/AIDS, sexual behavior, injury control, hypertension, cholesterol screening, and acculturation. Of the 660 adults interviewed, 364 (55.2%) were male and 296 (44.8%) were female. Among males, 31.9% were current smokers, and among females, only one woman reported smoking. Univariate analyses revealed that having less than a college education, having poor English language skills, using Vietnamese at home and with friends, being less acculturated, not having a routine physical or blood cholesterol check, and being a binge drinker were significantly associated with an increased likelihood of smoking. Multivariate analysis revealed two independently associated factors: Respondents who were more acculturated were less likely to smoke (OR = 0.38, 95% CI = 0.18-0.83), and those not having cholesterol checked were more likely to smoke (OR = 2.48, 95% CI = 1.30-4.71). Acculturation level was inversely associated with smoking among Vietnamese adult men in Santa Clara County. Other health risk behaviors coexisted with smoking behavior and should be considered in prevention programs. PMID- 15804683 TI - Voucher-based incentives for cigarette smoking reduction in a women's residential treatment program. AB - Participants were women (N = 16) living with their children in a residential substance abuse treatment facility. In this within-subjects repeated measures study, a 1-week baseline was followed by a 4-week intervention and a 2-week follow-up (same as the baseline). The intervention consisted of exposure to an educational video and a smoking cessation workbook, brief individual support meetings, and an escalating schedule of voucher-based reinforcement of abstinence. Throughout the study, three daily breath samples (8 a.m., noon, and 4 p.m.) were collected Monday through Friday to determine carbon monoxide (CO) concentration. In addition, urine cotinine (COT) was assessed on Monday mornings to monitor weekend tobacco use. Participants received vouchers of escalating value for CO-negative breath and COT-negative urine samples. Positive samples reset the voucher value. Significantly more negative tests were submitted during the intervention than during baseline and follow-up. The intensive behavioral intervention evaluated in this study produced a substantial reduction in cigarette smoking, and 25% of participants remained abstinent 2 weeks after the intervention was suspended. Nevertheless, the percentage of CO-negative samples submitted during the follow-up returned to baseline levels. While retaining many real-world characteristics, residential treatment facilities provide important opportunities for smoking cessation treatment and research. PMID- 15804684 TI - Nicotine patch and lozenge are effective for women. AB - It has been hypothesized that women may be less likely to obtain therapeutic benefit from nicotine replacement therapy (NRT). The present study tested this hypothesis, using two different types of NRT medications. A secondary analysis of two randomized clinical trials was performed: One compared active 21-mg nicotine patch with placebo among 193 men and 309 women, and the other compared active 2 mg or 4-mg nicotine lozenge with placebo among 788 men and 1,030 women. Using logistic regression analysis of 6-month continuous abstinence and survival analysis, we assessed the efficacy of patch and lozenge among women and tested for a gender x treatment interaction. Active NRT was more effective than placebo among women, for both patch and lozenge. In the lozenge trial, women were less successful than men. The gender x treatment interaction was not significant in either study, whether assessed by logistic regression or survival analysis. In the lozenge trial, gender moderated the effects of smoking rate and dependence (but not treatment) on outcome: These variables affected success rates only among women. Treatment with nicotine patch or lozenge is effective for women, and the analysis did not reveal significant gender differences in efficacy. Gender differences in outcome may be moderated by nicotine dependence. PMID- 15804685 TI - Dissemination of an effective inpatient tobacco use cessation program. AB - The present study aimed to determine whether tobacco use cessation rates observed in controlled trials of a hospital-based tobacco use cessation program could be replicated when the program was disseminated to a wide range of hospitals in a two-stage process including implementation and institutionalization phases. Using a nonrandomized, observational design, we recruited six hospitals to participate in the study. The research team helped implement the program during the first year of participation (implementation) and then withdrew from active involvement during the second year (institutionalization). The mean 6-month self-reported cessation rates were 26.3% (range = 17.6%-52.8%) for the implementation phase and 22.7% (range = 12.9%-48.2%) for the institutionalization phase. Hospitals with paid professionals providing the program had the best outcomes. Inpatient tobacco use cessation programs are feasible to implement and should target a 6-month self reported cessation rate of at least 25%. PMID- 15804686 TI - Effects of smoking cessation and reduction in asthmatics. AB - The present study examined the effect of smoking reduction and cessation on asthma regulation and biomarkers of exposure to cigarette smoke. In a prospective open design, we allocated 220 asthmatics among three groups: (a) Smoking reduction (reducers), with the aim of smoking fewer than seven cigarettes per day, (b) complete smoking cessation (abstainers), or (c) continuation of usual smoking (continuing smokers). Subjects used nicotine chewing gum or an oral nicotine inhaler to promote reduction and cessation. We monitored changes in the biomarkers carbon monoxide, cotinine, and thiocyanate, and in peak flow, medicine use, bronchial reactivity, and asthma symptoms. The analysis used the three outcome groups, regardless of original allocation to treatment groups. At 4 months, analysis of abstainers (n = 27), reducers (n = 33), and continuing smokers (n = 50) showed marked, statistically significant decreases in expired carbon monoxide of 17 ppm (abstainers) and 15 ppm (reducers); in plasma cotinine of 124 ng/ml (abstainers) and 122 ng/ml (reducers); and in plasma thiocyanate of 5.03 ng/ml (abstainers) and 3.74 ng/m (reducers). For abstainers, we observed improvements in the asthma-specific quality-of-life score, and reductions in self reported day and night use of rescue beta2-agonists, in doses of inhaled corticosteroids, in daytime asthma symptoms, and in bronchial hyperreactivity. For reducers, smaller improvements occurred for night use of rescue beta2 agonists, doses of inhaled corticosteroids, and bronchial hyperreactivity. Smoking cessation resulted in a marked decrease in three biomarkers of cigarette smoke inhalation and improved asthma regulation, whereas smoking reduction had a less pronounced effect on biomarkers and only a small effect on asthma regulation. PMID- 15804687 TI - Are waterpipe users interested in quitting? AB - Waterpipe smoking has increased dramatically worldwide in recent years, with an estimated 100 million daily users. Research on this traditional Middle Eastern tobacco use method is in its infancy, and little is known about users' cessation related attitudes and experiences. A random sample of 268 narghile (waterpipe) smokers (40% female; mean age = 30 years; range = 18-68) was obtained from cafes and restaurants in Aleppo, Syria. The majority of users (86.5%) believed they could quit using waterpipes at any time, but that belief was inversely related to perceived dependence, with only 48.7% of those who thought they were "very hooked" believing they could quit. Interest in quitting was expressed by 28.4% of subjects, with the majority (89.2%) reporting health concerns as a primary reason, and 59.2% having made an unsuccessful quit attempt in the past year. In a logistic regression model, independent predictors of interest in quitting included being married, having smoked for fewer years, not increasing the frequency of smoking over time, and having family members who do not smoke a waterpipe and disapprove of its use. Results indicate that a sizable percentage of waterpipe users express interest in quitting and have tried unsuccessfully in the past to quit. Waterpipe use needs to be considered in developing effective tobacco use cessation programs in the Middle East. PMID- 15804688 TI - Measuring adults' loss of autonomy over nicotine use: the Hooked on Nicotine Checklist. AB - The Hooked on Nicotine Checklist (HONC) is a 10-item screening tool originally developed to assess loss of autonomy over tobacco in adolescent smokers. A smoker's endorsement of any item indicates some loss of autonomy, and the sum of endorsed items indicates the degree to which autonomy has been lost. This study extends the HONC to adult smokers. Self-administered questionnaires were completed by a convenience sample of 1,102 adults who were recruited in their natural environments while observed smoking. Subjects were mostly experienced smokers who consumed a mean of 17.5 cigarettes per day and had smoked an average of 20.9 years. HONC scores ranged from 0 to 10, with a mean of 7.1. The HONC appears to measure a single dimension. Internal HONC consistency was high (alpha = .83), and interitem correlations were low to moderate. HONC scores correlated positively with levels of current and peak lifetime cigarette consumption. The HONC scores of heavy, moderate, and light smokers differed significantly in the expected direction. Subjects with higher HONC scores were more likely to report a shorter duration of abstinence and greater use of pharmacological aids when quitting. The HONC appears to be psychometrically sound in both adolescents and adults. It is a reliable and valid measure of lost autonomy for both novice and experienced smokers, and it allows for comparisons between these populations. PMID- 15804689 TI - Taking it to the streets: responses of African American young adults to internal tobacco industry documents. AB - Since the Master Settlement Agreement of 1998 between 46 states and the major tobacco companies forced the release of over 30 million pages of previously secret internal tobacco industry documents, researchers have been exploring how information in the documents can be useful for advancing public health efforts. Previous research shows that the tobacco industry has made massive efforts to target marginalized communities, not only through typical advertising channels but also through establishing financial and other ties with influential leadership groups. However, no previous studies have explored how members of targeted groups might respond at the grassroots level to actual internal tobacco industry documents about such targeting. This exploratory focus group study, which invited urban African American participants to comment on previously secret internal tobacco industry documents, suggests that such documents may be useful in efforts to socially denormalize tobacco use, promote critical reflection about community targeting, and mobilize individuals toward quitting. PMID- 15804693 TI - Women's status and health of two ethnic groups inhabiting a periurban habitat of Kolkata City, India: a micro-level study. AB - We studied the interrelationship of women's status in terms of socioeconomic inequality and its effect on women's health at micro level between two ethnic groups in a periurban area of Kolkata City, India. One-hundred twenty-seven women who belong to a tribal population (Munda) and 174 women who belong to a caste population (Poundrakshatriya) participated in this study. We found significant differences between various (socioeconomic, demographic, diet intake, and body mass index [BMI] factors among the two ethnic groups that indicated a better situation for the Pod women. The number of live births, dietary intake and BMI of the women of the two ethnic groups varied differentially among socioeconomic factors, such as women's education and working pattern and poverty level of the household, which are the most recognized measures of women's status. Thus, the diverse socioeconomic status in various cultural groups in traditional Indian societies reflects a more complex situation of women's status and their health. Different factors were responsible for the differential health status of women, which is culture and location specific. Women who are more educated and employed are not necessarily more healthy, since poverty remains an integral factor, base on which literacy and employment status of women in India is determined. Furthermore, suppression of women is rooted in the very fabric of the Indian society, in tradition, in religious doctrine and practices, within the educational systems, and within the families. Along with education, therefore, income-generating schemes for the women of the economically deprived population should be strengthened to bring equality in overall health status of a region that consists of diverse cultural populations with vast economic disparity. PMID- 15804694 TI - Perceived stress and support of the Chinese postpartum ritual "doing the month". AB - "Doing the month" is the traditional ritual for Chinese postpartum women. A qualitative study involving in-depth interviews with 20 Hong Kong Chinese women was conducted to examine the women's perceptions of stress and support in "doing the month." Four central themes were identified as follows: bound by the environmental constraints, difficulties in following the prescriptions of the rituals, conflicts between the parties involved, and attainment of the maternal role. The study revealed that "doing the month" is not necessarily protective and supportive of the postpartum women. It raises the critical question of how women can adapt the ritual to fit modern life. The study findings suggest that midwives and health care providers should provide culturally appropriate advice for new mothers. Suggestions are given on bridging the gap between traditional and modern postpartum practices. PMID- 15804695 TI - An integrative review on breast cancer screening practice and correlates among Chinese, Korean, Filipino, and Asian Indian American women. AB - Breast cancer is the leading cancer diagnosed among Chinese, Filipino and Korean women. Despite the efficacy of breast cancer screening, Asian American women have one of the lowest rates of mammography and clinical breast exam (CBE) of all U.S. ethnic groups. I provided a systematic overview to document factors related to three modalities of breast cancer screening (breast self-exam [BSE], CBE and mammography) among four subgroups of Asian American women (i.e., Chinese, Korean, Filipino, and Asian Indian). Results showed that the existing studies provided evidence of demographic correlates to screening practice; however, fewer studies focused on cognitive and social-cultural correlates, and limited results were available with Filipino and Asian Indian groups. PMID- 15804696 TI - Coping style and women's participation in breast and gynecological screening. AB - The purpose of this project was to determine the role of coping style in women's practice of breast and gynecological screening behaviors. Women were classified into one of four coping groups based on measures of vigilance and screening distress. Data were analyzed from a subset of 85 U.S. women who participated in a larger longitudinal study designed to examine self-reported screening participation. Results indicate frequency of screening participation and importance of obtaining immediate mammography feedback differed across coping style groups. Women characterized as high in vigilance and screening distress were most at risk for nonparticipation in screening. PMID- 15804697 TI - Assessment of risk factors for low birth weight deliveries. AB - Recognition of a higher than average incidence of low birth weight (LBW) deliveries in areas of Eastern Pennsylvania led to an analysis of more than 5,500 deliveries in one of the region's hospital systems to investigate the predictors of LBW in that population. Results of multivariate analysis indicate key variables that contribute to LBW: in particular, racial/ethnic background and specific medial problems during pregnancy, including preeclampsia, incompetent cervix, bleeding, low BMI and lack of adequate weight gain. Results were incorporated into training in best practices for prenatal care in the city's prenatal clinics. PMID- 15804698 TI - Autoantibodies-the smoke and the fire. PMID- 15804699 TI - Antinuclear antibodies. AB - The presence of abnormal levels of autoantibodies to intracellular antigens is a hallmark of systemic connective tissue disease. The indirect immunofluorescence assay is the most commonly used routine test for the detection of antinuclear antibodies. In this text, several representative patterns of fluorescence are reviewed and some pitfalls for application of the results to the clinical field are mentioned. PMID- 15804700 TI - Rheumatoid factor on a daily basis. AB - Rheumatoid factors (RF), which are antibodies (Ab) with specificity directed against gamma (?) globulins, are the commonest auto-Ab ever described in man. Some of them are referred to as agglutinating RF, others designated non agglutinating RF. Not only do these characterize rheumatoid arthritis (RA), but they are also encountered in a variety of disease conditions, as well as a proportion of healthy controls. Although non-specific for RA, the measurement of agglutinating IgM-RF remains the most useful serological test for the diagnosis of this disease. Demonstration of abnormal amount of serum RF by any method for which the result has been positive in less than 5% of normal subjects has indeed become one of the seven revised criteria, listed by the American College of Rheumatology (Arnett, FC, Edworthy, SM, Bloch, DA, McShane, DJ, Fries, JF, Cooper, NS, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum, 1988: 31: 315-24), for the classification of RA. Over the course of years, the relative importance of genetic (Carson, DA, Chen, PP, Kipps, TJ, Radoux, V, Jirik, FR, Goldfien, RD, et al. Idiotypic and genetic studies of human rheumatoid factors. Arthritis Rheum, 1987: 30: 1321-1325) and environmental (Nemazee, DA, Sato, VL. Enhancing antibody, a novel component of the immune response. Proc Natl Acad Sci USA, 1982: 79: 3828-3832) factors in the production of such intriguing auto-Ab has been delineated. PMID- 15804701 TI - Autoantibodies to citrullinated proteins: ACPA. AB - Anti-perinuclear factor and anti-keratin antibodies have long been known to be specifically associated with rheumatoid arthritis (RA). They were first demonstrated to target various forms of (pro)filaggrin, a protein of stratified epithelia. Then, they were found to belong to a single family of autoantibodies targeting proteins that bear peptidic epitopes centered by a citrullyl residue: the anti-citrullinated protein autoantibodies (ACPA). The main targets of ACPA in the synovial tissue were demonstrated to be citrullinated forms of the a- and beta-chains of fibrin. A chronic conflict between locally produced ACPA and deposits of citrullinated fibrin is probably responsible for self-maintaining of RA synovial inflammation. Various tests for the detection of ACPA have been developed: recent ELISAs confirm their high diagnostic specificity and improve their diagnostic sensitivity. Since ACPA appear very early in the course of the disease, their detection is of major interest to identify RA among recent arthritides. Moreover, their prognostic value may lead to start early 'aggressive' treatments to prevent irreversible joint damage. PMID- 15804702 TI - Autoantibodies to heterogeneous nuclear ribonucleoproteins. AB - Heterogeneous nuclear ribonucleoproteins (hnRNPs) are among the most abundant proteins in the eukaryotic cell nucleus and play a direct role in several aspects of the RNA life including splicing, export of the mature RNAs and translation. To date, approximately 30 proteins have been identified. A growing body of evidence points to hnRNPs as an important target of the autoimmune response in rheumatic diseases. Autoantibodies to A and B proteins of the hnRNP complex have been detected in late 1980s in rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD). Beyond their role as diagnostic test in clinical practice, these autoantibodies are starting to be regarded as important tools to obtain deeper insight into the pathogenesis of autoimmune rheumatic diseases. Furthermore, new anti-hnRNP antibodies have been recognized in the last ten years extending the spectrum of anti-hnRNP reactivity in different autoimmune disorders. PMID- 15804703 TI - Anticardiolipin and anti-beta-2-glycoprotein I antibodies. AB - The anticardiolipin (aCL) antibody test was first established in 1983, using cardiolipin (negatively charged phospholipid) as an antigen in a solid-phase immunoassAy. It was first applied to the study of systemic lupus erythematosus patients, and was found associated with thromboses and recurrent pregnancy losses. The wide use of this test was determinant in the definition of the "aCL or antiphospholipid syndrome" (APS).Later, it was demonstrated that aCL antibodies do not recognize anionic phospholipids but are directed against plasma proteins bound to anionic phospholipids, mainly beta-2-glycoprotein I, which is now considered as the autoantigen in APS. Anti-beta-2-glycoprotein I (anti beta2GPI) is not yet accepted as a serological criterion for APS, but most investigators would consider a patient with anti-beta2GPI antibodies and clinical features of APS to have the syndrome. aCL and anti-beta2GPI are a heterogeneous group of antibodies with different clinical significances and can be present in different autoimmune diseases as well as in infectious diseases. PMID- 15804704 TI - Anti-DNA antibodies: a diagnostic and prognostic tool for systemic lupus erythematosus? AB - The clinical impact of anti-DNA antibodies lies on their diagnostic power for systemic lupus erythematosus (SLE), being a formal classification criterion. In spite of such a disease association, low-avidity anti-DNA antibodies might also be part of the natural autoantibody repertoire. Their switch to pathogenic high avidity autoantibodies is the result of the autoimmune process leading to SLE.Anti-DNA antibodies were shown to play a role in SLE pathogenesis and particularly in kidney damage. Accordingly, antibody titres might fluctuate in relation to disease activity, but their prognostic value for flares is still debated.Several methods for anti-DNA detection were described and there is evidence that the assays identify different antibodies with different prognostic value. The results of a multicenter study on four different routine tests for anti-dsDNA antibody detection showed that: (i) Farr assay displays the best diagnostic specificity/sensitivity for SLE, followed by Crithidia luciliae method (CLIFT), (ii) the new generation of solid phase assay (EliA) shows an increased sensibility versus the classical enzyme linked immune assay (ELISA) but a decreased specificity. Antibody titre detected by EliA and Farr assay correlated with disease activity. These findings would suggest that more than one assay should be useful for SLE diagnosis and monitoring. PMID- 15804705 TI - Anti-Sm and anti-RNP antibodies. AB - Among anti-nuclear antibodies, anti-Sm and anti-RNP antibodies are of the utmost importance in clinical practice. Anti-Sm antibodies are directed against 7 proteins (B/B', D1, D2, D3, E, F, G) that constitute the common core of U1, U2, U4 and U5 small nuclear ribonucleoprotein (snRNP) particles; B/B', D1 and D3 are more frequently targeted. Anti-RNP antibodies react with proteins (70 Kd, A, C) that are associated with U1 RNA and form U1snRNP. Anti-Sm and anti-RNP antibodies are directed towards both discontinuous and linear epitopes which are either contained in the protein sequence or are post-translationally modified. The assays to detect anti-Sm and anti-RNP antibodies are counterimmunoelectrophoresis (CIE), immunoblot, and ELISA, based on purified or recombinant proteins or synthetic peptides. Anti-Sm antibodies are detectable in a percentage of SLE patients comprised between 5 and 30%; they are more prevalent in blacks and because of their high specificity for SLE have been included in the serological criteria for diagnosing the disease.Anti-RNP are detectable in 25-47% of SLE patients; high titers of anti-RNP antibodies are diagnostic of mixed connective tissue disorder (MCTD). The measurement of anti-Sm and anti-RNP antibodies is more important in the diagnosis of SLE than in the follow-up of patients. However, anti-RNP antibodies are more prevalent in patients with Raynaud's phenomenon and are associated with milder renal involvement. On the contrary, anti-Sm antibodies are associated with the severity and the activity of renal involvement. The specificity of anti-Sm antibodies, together with epidemiological data, suggest that Epstein-Barr virus infection has the potential to induce anti Sm antibodies by molecular mimicry.Anti-nuclear antibodies, a hallmark of the systemic autoimmune diseases, include several populations of antibodies with different specificities. Among them, anti-Sm and anti-RNP antibodies are of the utmost importance in clinical practice; in research, the study of the mechanisms inducing their production has opened up new perspectives and helped to elucidate the pathogenesis of autoimmune disorders. PMID- 15804706 TI - Anti-Ro/SSA and La/SSB antibodies. AB - The Ro/La system is considered as an heterogeneous antigenic complex, constituted by three different proteins (52 kDa Ro, 60 kDa Ro and La) and four small RNAs particles. Anti-Ro/SSA are the most prevalent specificity among many autoimmune diseases, such as systemic lupus erythematosus (SLE), SS/SLE overlap syndrome, subacute cutaneous LE (SCLE), neonatal lupus and primary biliary cirrhosis. In contrast, anti-La/SSB is more associated with Sjogren's syndrome (SS). The differences between 52 kDa, 60 kDa Ro and La could explain why different assays did not show equivalent performance in anti-Ro and anti-La autoantibodies detection. The RNA precipitation assay had the highest sensitivity and specificity, usually considered as the reference methods. CIE is considered the most reliable to detect anti-Ro/SSA antibodies in routine practice, performing better than immunoblotting (IB) and some ELISAs. It shows a high sensitivity (89%) and specificity (100%). ELISA is generally considered a safe, rapid, sensitive and specific tecnique. Therefore, its high sensitivity often corresponds to a very low clinical specificity and the assay can give false positive results. Therefore, it is very important to search anti-Ro and anti-La only in selected patients, using the assay with high specificity and good predictive value, in order to have clinically significant and true positive results. PMID- 15804707 TI - Anti-scl-70. AB - OBJECTIVE: To develop an overview focusing on the utility of anti-Scl-70 autoantibody determinations in the rheumatic diseases. METHODS: Articles from electronic literature searches were retrieved, critiqued and data were extracted and pooled on anti-Scl-70 (topoisomerase I) in relation to history, optimal tests used for its detection, sensitivity, specificity, positive and negative likelihood ratios, indications, interpretation and pitfalls. RESULTS: Anti-Scl 70 antibodies are very useful in distinguishing systemic sclerosis (SSc) patients from healthy controls, from patients with other connective tissue diseases, and from unaffected family members. Among patients with SSc, anti-Scl-70 positivity is useful in predicting those at higher risk for diffuse cutaneous involvement and interstitial fibrosis/restrictive lung disease, though the latter has not been universally observed. Of the four different techniques notably immunodiffusion, immunoblotting, immunoprecipitation and enzyme-linked immunosorbent assay (ELISA) used to assay anti-Scl-70, immunodiffusion has been the most extensively validated. ELISAs are somewhat less specific than other techniques, especially in distinguishing SSc patients from those with other rheumatic diseases, though newer generation ELISAs have been developed to overcome the problem of low specificity inherent with the traditional techniques. As of yet, the need for serial testing of anti-Scl-70 has not been established. CONCLUSIONS: Evidence-based guidelines suggest that anti-Scl-70 antibodies are very useful in the diagnosis and clinical management of SSc patients and also to establish prognosis in these patients, particularly those with diffuse skin involvement. PMID- 15804708 TI - Anti-Jo-1 antibodies. AB - Anti-Jo-1 antibody is a myositis specific autoantibody most commonly found in patients with idiopathic inflammatory myopathies (IIM). This antibody is directed against the histidyl-tRNA synthetase which catalyses the binding of the histidine to its cognate tRNA during protein synthesis. It can be considered a specific marker of IIM, predominantly found in 20-30% of patients with PM and in the 60 70% of those with interstitial pulmonary fibrosis. These antibodies are also found in DM, although less frequently than in PM, and are rare in children with PM or DM and in other connective tissue diseases.ELISA, CIE and immunoblotting are highly specific and sensitive techniques for testing anti-Jo-1 antibodies. The detection of this antibody is particularly useful in diagnosis and classification of IIM. Moreover, anti-Jo-1 serum levels strongly correlate with disease activity representing a good marker for disease monitoring. PMID- 15804709 TI - Anti-ribosomal P protein antibodies. AB - Anti-ribosomal P protein antibodies (Abs) recognize three specific ribosomal proteins located in the large ribosome's subunit. The term anti-ribosomal P protein Abs, often shortened in anti-P Abs, is due to the fact that these three proteins, P0, P1 and P2, of 38, 19 and 17 Kd molecular weight, respectively, are phosphorylated. One of the major points of interest of these autoAbs derives from their high specificity for systemic lupus erythematosus (SLE), their description in other connective tissue diseases (CTDs) being only occasional. In SLE patients, their association with renal and hepatic involvement has been proposed, while the possible association with psychiatric and/or neurological involvement is still a matter of debate. From a serological point of view, a preferential association of anti-P with anti-Sm and/or anti-DNA Abs, possibly due, at least in part, to cross-reactivity, has been suggested. This observation is intriguing since all these autoAbs are considered specific serological indicators of SLE. This review will summarize clinical and serological data on anti-P Abs provided by the main studies published in the last few years and the more recent findings about proteins constituting their targets. PMID- 15804710 TI - Antineutrophil cytoplasmic antibodies (ANCA). AB - Antineutrophil cytoplasmic antibodies (ANCA) are a sensitive and specific marker for ANCA-associated systemic vasculitis. Using indirect immunofluorescence on ethanol-fixed neutrophils, two major fluoroscopic patterns can be recognised: a diffuse cytoplasmic staining (C-ANCA), and a perinuclear/nuclear staining (P ANCA). In patients with vasculitis, more of 90% of C-ANCA are directed against proteinase 3 (PR3-ANCA) whereas approximately 80-90% of P-ANCA recognise myelperoxidase (MPO-ANCA). Although C-ANCA (PR3-ANCA) is preferentially associated with Wegener's granulomatosis (WG), and P-ANCA (MPO-ANCA) with microscopic polyangiitis (MPA), idiopathic necrotising crescentic glomerulonephritis (iNCGN) and Churg-Strauss syndrome (CSS), there is not absolute specificity. Between 10-20% of patients with classical WG show P-ANCA (MPO-ANCA), and even a larger percentage of patients with MPA or CSS have C-ANCA (PR3-ANCA). Furthermore, it should be stressed that approximately 10-20% of patients with WG or MPA (and 40-50% of cases of CSS) have negative assay for ANCA. The best diagnostic performance is obtained when indirect immunofluorescence is combined with PR3 and MPO-specific ELISAs. ANCA with different and unknown antigen specificity are found in a variety of conditions other than AASV, including inflammatory bowel diseases, other autoimmune diseases, and infections where their clinical significance is unclear. ANCA levels are useful to monitor disease activity but should not be used by themselves to guide treatment. A significant increase in ANCA titres, or the reappearance of ANCA, should alert the clinicians and lead to a stricter patient control. PMID- 15804711 TI - Multiplexed AtheNA multi-lyte immunoassay for ANA screening in autoimmune diseases. AB - BACKGROUND: Multiplexed assays using fluorescence microspheres is an exciting technology with multiple applications including the detection of antinuclear autoantibodies (ANA) and autoantibody profiles. It is a rapid, sensitive and automatic method for simultaneous quantitative detection of several autoantibodies. The aim of our study was to determinate ANA and other autoantibodies to the nine extractable nuclear antigens by the AtheNA Multi-Lyte ANA system and compare the results achieved by this method to the routinely used enzyme immunoassay. METHODS: Four hundred eighteen serum samples were tested utililizing the multiplexed method: 96 healthy donors, 86 requested ANA specimens obtained from routine lab, and 236 samples from patients with known autoimmune diseases (43-scleroderma, 113-systemic lupus erythematosus, 38-Sjogren's syndrome, and 42 rheumatoid arthritis). The ANA and antibodies to nine different analytes (SS/A, SS/B, Sm, RNP, Jo-1, Scl-70, dsDNA, Centromere B and Histone) were tested. RESULTS: ANA screening by AtheNA system revealed high concordance of 99 and 97.7% with the enzyme immunoassay test in samples obtained from healthy donors and ANA requested samples, respectively. Evaluation of autoimmune disease related samples for ANA by AtheNA technology also confirmed a high rate of concordance of 92-97.7% and correlated with the enzyme immunoassay. Positive discrepant results were found for Scl-70 specificity in 12.7% of SLE specimens by AtheNA technology, while all tested sera were negative for this antibody by enzyme immunoassay. Negative discrepant results were observed by the AtheNA system for anti-dsDNA. The sera (15 randomly obtained samples from SLE patients) were positive for anti-dsDNA in 50% of samples in Farr assay and 55% in enzyme immunoassay, respectively. CONCLUSION: We suggest that the AtheNA technology may be a useful diagnostic tool for ANA screening. Additional investigations are required to compare an analytic performance between AtheNA and routine methods in determination of the individual autoantibody profile. PMID- 15804712 TI - Surface elastic modulus of barnacle adhesive and release characteristics from silicone surfaces. AB - The properties of barnacle adhesive on silicone surfaces were studied by AFM indentation, imaging, and other tests and compared to the barnacle shear adhesion strength. A multilayered structure of barnacle adhesive plaque is proposed based on layered modulus regions measured by AFM indentation. The fracture of barnacles from PDMS surfaces was found to include both interfacial and cohesive failure of barnacle adhesive plaque, as determined by protein staining of the substratum after forced barnacle release from the substrate. Data for freshly released barnacles showed that there was a strong correlation between the mean Young's modulus of the outermost (softest) adhesive layer (E<0.3 MPa) and the shear strength of adhesion, but no correlation for other higher modulus regions. Linear, quadratic, and Griffith's failure criterion (based on rough estimate of crack length) regressions were used in the fit, and showed significance. PMID- 15804713 TI - Recruitment in the field of Balanus improvisus and Mytilus edulis in response to the antifouling cyclopeptides barettin and 8,9-dihydrobarettin from the marine sponge Geodia barretti. AB - In this field investigation the two cyclopeptides, isolated from the marine sponge Geodia barretti Bowerbank (Geodiidae, Astrophorida), are shown to be very efficient in preventing recruitment of the barnacle Balanus improvisus (Cirripedia, Crustacea) and the blue mussel Mytilis edulis (Protobranchia, Lamellibranchia) when included in different marine paints. These brominated cyclopeptides, named barettin and 8,9-dihydrobarettin were incorporated in different non-toxic coatings. The substances were used in the concentrations 0.1 and 0.01% in all treatments. The most efficient paint was a SPC polymer. This paint, in combination with barettin and 8,9-dihydrobarettin, reduced the recruitment of B. improvisus by 89% (barettin, 0.1%) and by 67% (8,9 dihydrobarettin, 0.1%) as compared to control panels. For M. edulis, the reduction of recruitment was 81% with barettin (0.1%) and 72% with 8,9 dihydrobarettin (0.1%) included in the SPC paint. This indicates that the two compounds from G. barretti could provide non-toxic alternatives as additives in antifouling paints, since the heavy metal-based marine paints are to be replaced. PMID- 15804714 TI - Activity of commercial enzymes on settlement and adhesion of cypris larvae of the barnacle Balanus amphitrite, spores of the green alga Ulva linza, and the diatom Navicula perminuta. AB - Fouling species produce adhesive polymers during the settlement, adhesion and colonization of new surfaces in the marine environment. The present paper tests the hypothesis that enzymes of the appropriate specificity may prevent biofouling by hydrolysing these adhesive polymers. Seventeen commercially available enzyme preparations designed originally for bulk use in a range of end-use applications were tested for their effects on the settlement and/or adhesion of three major fouling species, viz. the green alga Ulva linza, the diatom Navicula perminuta and the barnacle Balanus amphitrite. The serine-proteases were found to have the broadest antifouling potential reducing the adhesion strength of spores and sporelings of U. linza, cells of N. perminuta and inhibiting settlement of cypris larvae of B. amphitrite. Mode-of-action studies on the serine-protease, Alcalase, indicated that this enzyme reduced adhesion of U. linza in a concentration dependent manner, that spores of the species could recover their adhesive strength if the enzyme was removed and that the adhesive of U. linza and juvenile cement of B. amphitrite became progressively less sensitive to hydrolysis as they cured. PMID- 15804715 TI - Adrenoceptor compounds prevent the settlement of marine invertebrate larvae: Balanus amphitrite (Cirripedia), Bugula neritina (Bryozoa) and Hydroides elegans (Polychaeta). AB - The effects of the neurotransmitter blockers idazoxan and phentolamine on the larval settlement of three marine invertebrate species belonging to three different phyla were investigated by using in vitro concentration-response bioassays. Since neurotransmitters are known to influence metamorphic transitions in invertebrate larvae, neurotransmitter blockers were tested to evaluated their sublethal effects on larvae. The alpha-adrenergic antagonists idazoxan and phentolamine inhibited settlement of Balanus amphitrite (Cirripedia), Bugula neritina (Bryozoa) larvae, and larvae of the polychaete Hydroides elegans (Polychaeta) in a concentration-and taxon-dependent manner. At concentrations of 10(-3) M of both agents, larvae of all three species became immobile and subsequently died within 24 h. While cumulative settlement rates were observed after 48 h for B. amphitrite and H. elegans, and after 5 h for B. neritina, >90% of the larvae that settled did so within 24 h for the first two species and within 1 h for B. neritina. The tendency of the hydrophobic idazoxan and phentolamine to accumulate at solid surfaces most probably contributes to their successful inhibition of larval settlement. This ability makes them particularly attractive as candidates for the development of slow-release carriers in antifouling paints. PMID- 15804716 TI - Adhesion and motility of fouling diatoms on a silicone elastomer. AB - Recent demands for non-toxic antifouling technologies have led to increased interest in coatings based on silicone elastomers that 'release' macrofouling organisms when hydrodynamic conditions are sufficiently robust. However, these types of coatings accumulate diatom slimes, which are not released even from vessels operating at high speeds (>30 knots). In this study, adhesion strength and motility of three common fouling diatoms (Amphora coffeaeformis var. perpusilla (Grunow) Cleve, Craspedostauros australis Cox and Navicula perminuta Grunow) were measured on a poly-dimethylsiloxane elastomer (PDMSE) and acid washed glass. Adhesion of the three species was stronger to PDMSE than to glass but the adhesion strengths varied. The wall shear stress required to remove 50% of cells from PDMSE was 17 Pa for Craspedostauros, 24 Pa for Amphora and >>53 Pa for Navicula; the corresponding values for glass were 3, 10 and 25 Pa. In contrast, the motility of the three species showed little or no correlation between the two surfaces. Craspedostauros moved equally well on glass and PDMSE, Amphora moved more on glass initially before movement ceased and Navicula moved more on PDMSE before movement ceased. The results show that fouling diatoms adhere more strongly to a hydrophobic PDMSE surface, and this feature may contribute to their successful colonization of low surface energy, foul-release coatings. The results also indicate that diatom motility is not related to adhesion strength, and motility does not appear to be a useful indicator of surface preference by diatoms. PMID- 15804717 TI - Effects of ischemia-reperfusion on NMDA receptor subunits 2a and 2b level in rat hippocampus. AB - The authors investigated the effects of ischemia and reperfusion on the N-methyl D-aspartate receptor (NMDAR) subunits 2A and 2B concentration in rat hippocampus. At the protein level, significant increase in the amounts of NMDAR 2A and NMIDAR 2B in the rat hippocampus was observed at 1 h after reperfusion compared with control group. These results suggested that the alteration in hippocampal NMDAR2 subunit concentrations after ischemia-reperfusion might be invovlved in cognitive dysfunction and excitotoxicity. PMID- 15804718 TI - The relationship of HRV to sleep EEG and sleep rhythm. AB - Previous studies have shown that there exists a cycle of NREM (non-rapid eye movement)-REM (rapid eye movement) during normal human sleep, and heart rate variability (HRV) has a close relationship to sleep stages and sleep cycle. This article reports the relationship between the electroencephalographic activity and the HRV spectral power in several specific frequency bands. The authors discovered that relationships do exist between HRV and electroencephalogram (EEG) during sleep. In particular, it was found that, prior to the changes of EEG, the changes of HRV usually indicate the shift of sleep stages. HRV frequency analysis indicates that the very-low-frequency components of HRV are closely related to sleep EEG. Results show that the rhythm of the spectral power oscillations in some specific frequency bands of HRV is almost the same as the sleep cycle, which reflects the rhythm of sleep to a certain extent. PMID- 15804719 TI - Magnetic storage of information in the human cerebral cortex: a hypothesis for memory. AB - The diversity of memory phenomena argues against a single place or anatomical structure for memory in the nervous system. Moreover, molecular mechanisms of information storage and synaptic transmission seem insufficient to support contextual recall and other very complex human memory processes. Here, we propose a new physical model for memory based on the magnetic fields associated with neuronal activity and its possible interaction with the adjacent astroglial network. The hypothesis emphasizes the architectural organization of the human cerebral cortex because the close geometrical relationships between neuronal minicolums and astroglial network acquire transcendental importance. PMID- 15804720 TI - Effects of tramadol on nerve action potentials in rat: comparisons with benzocaine and lidocaine. AB - The effects of tramadol on repetitively elicited action potentials were studied in rat sciatic nerve, using the sucrose gap method. Tramadol's local anesthetic like effects were compared with lidocaine and benzocaine at single or 10, 40, and 100 Hz stimulations. Tramadol and lidocaine both produced approximately the same level of conduction block. The depolarization time of the compound action potentials (CAP) measured from the beginning to the peak of the CAPs, was extended by lidocaine and tramadol, but benzocaine had no effect in this respect. Tramadol extended half width of CAP more than lidocaine. Lidocaine and tramadol produced similar conduction-block patterns, which were different from benzocaine. The results suggested that tramadol enhanced the nerve conduction like lidocaine. However, their frequency-dependent block patterns were similar. It was concluded that tramadol may block the Na+ channels following the hydrophilic pathway like lidocaine and block K+ channels more than lidocaine. These may accounted for the local anesthetic-like effects of tramadol. PMID- 15804721 TI - Working memory processing of facial images in schizophrenia: fMRI investigation. AB - Impaired processing of facial information is one of the broad ranges of cognitive deficits seen in patients with schizophrenia. This study aimed to elucidate the differences in brain activities involved in the process of facial working memory (images of neutral faces as mnemonic content) between schizophrenic patients and healthy comparison subjects using functional magnetic resonance imaging (fMRI). Group comparison analysis revealed aberrant activities from the patient group in several cortical areas, including the left fusiform gyrus, right superior frontal gyrus, bilateral middle frontal gyri/insula, and left middle temporal gyrus. A decreased level of activity in the left fusiform gyrus, along with performance reduction, implicates abnormal processing of facial information in schizophrenia. PMID- 15804722 TI - Effects of fluvoxamine on levels of dopamine, serotonin, and their metabolites in the hippocampus elicited by isolation housing and novelty stress in adult rats. AB - The authors investigated the effects of fluvoxamine on neurochemical changes in the hippocampus elicited by isolation housing and novelty stress. Male F344 rats (11 w) were housed one per cage for four weeks. On each day of the last week (7 days) they were s.c. injected with fluvoxamine (20 mg/kg), and then subjected to novelty stress. Isolation housing significantly increased dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA) levels, whereas fluvoxamine significantly decreased them. Isolation housing significantly increased the DOPAC/DA ratio. Fluvoxamine significantly decreased the DA level, and partially restored the DOPAC and 5-HIAA levels increased by isolation housing. PMID- 15804723 TI - Reversal of neuromuscular effects of adenosine by specific adenosine A1-receptor antagonist in live rats. AB - Intravenous adenosine in-vivo was shown to potentiate the effects of non depolarizing neuromuscular blocking agents. This study aimed to determine whether adenosine A1-receptors mediated this potentiation. The authors investigated the effects of intravenous adenosine, N6-cyclopentyladenosine, specific A1-receptor agonist, and 8-cyclopentyl-1,3-dipropylxanthine, specific A1-receptor antagonist, on neuromuscular block by vecuronium, in in-vivo rat sciatic nerve-tibialis anterior preparations. In the presence of 50% steady state block by vecuronium, adenosine, and N6-cyclopentyladenosine caused similar degree of depressions of twitch tension. Twitch tension returned to its pre-injection value more rapidly when 8-cyclopentyl-1,3-dipropylxanthine was given at the maximal block than when it was allowed to recover spontaneously. It was concluded that in in-vivo adenosine potentiated the neuromuscular effects of vecuronium through adenosine A1-receptors in rats. PMID- 15804724 TI - Morphological recovery of the granule cells from the olfactory bulb after the cessation of acute ozone exposure. AB - The purpose of this study was to analyze the possible morphological recovery of the granule cells in the olfactory bulb as a consequence of oxidative stress after an acute ozone exposure. Rats were divided in two groups: Control (air exposed) and experimental group, exposed 4 h, to 1 ppm ozone and divided into 4 subgroups, which were sacrificed at 2 and 24 h, 10 and 15 days, respectively. Olfactory bulbs were processed with the rapid Golgi method and for transmission electron microscopy. The granule cells of the olfactory bulb disclosed less dendritic spine density at 2, 24 h, and 10 days after the exposure compared with controls. At 15 days, the number of spines increased to values similar to those found in controls. The granule cells ultrastructure demonstrated an increment in lipofucsin granules, as well as swollen organelles, changes that decreased overtime. This change decline might be related to a partial recovery of the associative granule cells function. PMID- 15804725 TI - Dopamine cell morphology and glial cell hypertrophy and process branching in the nigrostriatal system after striatal 6-OHDA analyzed by specific sterological tools. AB - Morphological changes in the dopamine neurons and glial cells of the rat mid brain ascending dopamine pathways were investigated after a partial lesion induced by unilateral striatal injection of a small dose of 6-hydroxydopamine (6 OHDA). Fourteen days after lesion, animals showed contralateral rotation induced by apomorphine injection. After behavioral analysis, fats were killed and their brains processed for the immunohistochemistry tyrosine hydroxylase (TH), a marker for dopamine cells, as well as glial fibrillary acidic protein (GFAP) and OX-42, markers for astrocyte and microglia, respectively. Stereological tools were employed in the quantifications. The volumes of the regions of the striatal TH immunoreactive disappearance, as well as the astroglial and microglial activation were several folds increased compared to control saline-injected rats. The optical disector detected decreases in the estimated total number of dopamine cells in the entire ipsilateral pars compacta of the substantia nigra (SNc) and the ventral tegmental area (VTA) as well as in the estimated total number of varicosity profiles in the entire ipsilateral neostriatum. The stereological tool rotator showed no changes either in the mean or in the histogram distribution of the cytoplasmic volume of the nigral and VTA dopamine cells of 6-OHDA lesioned rats. Increases in the estimated total number of GFAP positive astrocytes were found in the entire neostriatum bilaterally as well as in the ipsilateral entire SNc and VTA of 6-OHDA lesioned rats. The estimated total number of OX-42 immunoreactive microglial profiles was elevated only in the ipsilateral entire neostriatum of the lesioned rats. The rotator detected cytoplasmic hypertrophy in the astrocytes, and also a shift to the fight of the gaussian curves of the normal distribution of the logarithmic plotted values of the astroglial cell body volumes of the neostriatum bilaterally as well as in the ipsilateral SNc and VTA of the striatal 6-OHDA injected rats. Cytoplasmic hypertrophy of microglia, and also a shift to the right of the gaussian curves of the values of microglia cell body volumes were seen only in the ipsilateral neostriatum; however, the point intercepts revealed an increased amount of microglial processes in the ipsilateral SNc and VTA of the lesioned rats. Specific stereological methods can be applied on detection of regionally different forms of cellular astroglial and microglial reaction after a partial lesion of dopamine pathway. PMID- 15804726 TI - Towards safer women, safer doses, safer routes and safer timing of administration of safer menopausal therapies. PMID- 15804727 TI - KEEPS: The Kronos Early Estrogen Prevention Study. AB - Observational studies have indicated that hormone therapy given at or after menopause is linked to substantial reduction in cardiovascular disease and its risk factors. Recent findings from the Women's Health Initiative (WHI) clinical trial, however, indicate that combined estrogen plus progestin hormone therapy, as well as estrogen-alone hormone therapy (given to women without a uterus), is ineffective in preventing the new onset of cardiac events in previously healthy late menopausal women. Further, the secondary prevention trial, the Heart and Estrogen/progestin Replacement Study (HERS), also failed to demonstrate any benefit of initiation of hormone therapy in women with established coronary heart disease. In light of these results, a hypothesis has arisen that early initiation of hormone therapy, in women who are at the inception of their menopause, will delay the onset of subclinical cardiovascular disease in women. The rationale that earlier intervention than that performed in the WHI and HERS trials will provide cardiovascular benefit to women is the driving force behind the Kronos Early Estrogen Prevention Study, or KEEPS. KEEPS is a multicenter, 5-year clinical trial that will evaluate the effectiveness of 0.45 mg of conjugated equine estrogens, 50 microg weekly transdermal estradiol (both in combination with cyclic oral, micronized progesterone, 200 mg for 12 days each month), and placebo in preventing progression of carotid intimal medial thickness and the accrual of coronary calcium in women aged 42-58 years who are within 36 months of their final menstrual period. A total of 720 women are planned to be enrolled in 2005, with an anticipated close-out of the trial in 2010. This overview summarizes the recruitment and methodology of the KEEPS trial. PMID- 15804728 TI - Effects of low-dose hormone therapy on menopausal symptoms, bone mineral density, endometrium, and the cardiovascular system: a review of randomized clinical trials. AB - OBJECTIVES: First, to determine the extent of the effects of low-dose hormone therapy (HT) on menopausal symptoms, bone mineral density, endometrium, and the cardiovascular system, and, second, to determine the adverse effects of low-dose HT. METHODS: A literature review of electronic databases was conducted to identify all prospective, randomized trials comparing the effects of low-dose HT with placebo or standard-dose therapy, using key words such as: hormone replacement therapy (HRT), low-dose HRT/conjugated equine estrogens (CEE)/estradiol, lower-dose HRT/CEE/estradiol, ultra-low-dose HRT/CEE/estradiol, menopause, cardiovascular risk, bone metabolism. RESULTS: Low-dose HT has been shown to improve menopausal and vulvovaginal atrophic symptoms, compared to placebo, and is less likely to give rise to unacceptable side-effects, including irregular bleeding and/or breast tenderness. When compared to standard-dose HT, the low-dose HT has comparable effects on a range of menopausal symptoms and on bone density and has similar beneficial effects on surrogate end-points of coronary heart disease. CONCLUSIONS: A change to low-dose HT has been advocated following adverse findings in recent trials of standard-dose HT. Although a literature review has shown low-dose HT to alleviate menopausal symptoms and maintain or improve bone density with fewer side-effects than standard-dose therapy, further research is required to determine what effect the lower-dose therapy will have on fracture, cardiovascular and breast disease. PMID- 15804729 TI - Women's perceptions of hormone replacement therapy: risks and benefits (1980 2002). A literature review. AB - OBJECTIVES: Many postmenopausal women are reluctant to take hormone replacement therapy (HRT) and those who do are likely to discontinue within 1 year. Setting aside the beliefs and prejudices of the prescriber, women's own perceptions of risks and benefits may influence their willingness to accept and then persevere with HRT. We conducted a systematic literature review relating women's beliefs about HRT to acceptance and adherence. METHODS: Using standard guidelines for systematic search procedures, we identified 112 papers (published during 1980 2002) from databases such as Medline, PsycINFO and the NHS and Cochrane libraries. RESULTS: Women hold both positive and negative beliefs about HRT. Their perceptions of benefits are often countered by concerns over potential adverse effects, beliefs that are sometimes at odds with the clinical evidence. The use and discontinuation of HRT are influenced more by short-term symptom relief than by considerations of long-term benefits. Many women who refuse HRT believe that the menopause is a natural event that does not warrant 'chemical' intervention. Doctors and nurses are not used to their full capacity as an education and information resource to counter the possibility of unbalanced stories in the media. CONCLUSIONS: Addressing women's preconceptions about HRT should be an important part of the process of prescribing and review. Such an approach will help ensure that a woman's decision to start or continue HRT is informed by an understanding of the known risks and benefits, rather than by myths or mistaken beliefs about the menopause or HRT. PMID- 15804730 TI - Relevance of the results of the Women's Health Initiative on the prescription of hormone therapy in Spain. AB - OBJECTIVE: To identify the possible repercussions of suspending the estrogen gestagen arm of the WHI trial among Spanish health-care professionals habitually using hormonal therapy in their practice of medicine and to attempt to identify the possible changes in their attitudes and prescription habits regarding this issue. DESIGN: We designed a survey in the form of an anonymous self-administered interview with 18 questions, distributed to 3592 specialists in Obstetrics and Gynecology belonging to the Spanish Association for the Study of the Menopause (AEEM in its Spanish acronym) and the Spanish Gynecology and Obstetrics Society (SEGO in its Spanish acronym). The study was national in scope and the questionnaires were distributed between November 2002 and January 2003, with reception of questionnaires concluding in February 2003. RESULTS: Of the participants, 96% stated that they knew the results of the Women's Health Initiative (WHI) study as published in the Journal of the American Medical Association; 63% felt that the cardiovascular results of the trial were important for their clinical practice (55% of males and 80% of females surveyed), and 42% of the men and 18% of women stated the opposite. The results of the WHI should not be extrapolated to other types of hormone therapy (84%) and only 10% considered this possibility acceptable. CONCLUSIONS: From our study, it can be inferred that Spanish gynecologists believe they know the WHI study and state that its results cannot be extrapolated to our setting. However, following its publication, they have suspended more than 10% of therapies, have limited their indications, and have reduced their recommended duration. PMID- 15804731 TI - Self-reported arthritis and the menopause. AB - OBJECTIVES: To determine the prevalence of self-reported arthritis in a population-based cohort of mid-aged women and to identify health, social and lifestyle factors associated with self-reported arthritis. DESIGN: Cross sectional population-based survey. METHOD: Data were obtained from a telephone administered questionnaire of 2001 Australian-born women between 45 and 55 years old and residing in Melbourne. Information was collected on demographics, lifestyle factors and health conditions. RESULTS: The most commonly reported symptom in this cohort was aches and stiff joints, reported by 51.7% of the women; 34.4% of participants reported they had been diagnosed with arthritis. More postmenopausal women (39%) reported that they were diagnosed with arthritis compared to premenopausal women (27%, p < 0.001). Women who reported arthritis were more likely to be older (odds radio (OR), 1.09; 95% confidence interval (CI), 1.05-1.13), have a higher body mass index (OR, 1.04; CI, 1.02-1.07), be postmenopausal (OR, 1.88; CI, 1.33-2.66), have a higher negative mood (OR, 1.80; CI, 1.13-2.87), and report a decreased interest in sex (OR, 1.58; CI, 1.26-1.97). CONCLUSIONS: Aches and stiff joints are the most frequently reported symptom of mid-aged women. Reported arthritis is associated with postmenopausal status, age, body mass index, less interest in sex and lowered mood. Further longitudinal research is needed to determine the role of these factors in the development and impact of arthritis. PMID- 15804732 TI - Greene Climacteric Scale: norms in an Australian population in relation to age and menopausal status. AB - OBJECTIVES: The aim of this study was two-fold: to assess climacteric symptoms and provide normative data for the Greene Climacteric Scale during the menopause transition, and to investigate the prevalence of climacteric symptoms in a representative sample of postmenopausal Australian women. METHOD: A cohort of 500 premenopausal, perimenopausal and postmenopausal women aged 40-80 years participated in the Longitudinal Study of Ageing in Women (LAW study) at the Royal Brisbane and Women's Hospital, Brisbane, Australia. In year 1 of the study (2001), all participants completed the Greene Climacteric Scale and information regarding their menopausal status and the use of hormone therapy (HT) was obtained through a clinical interview with a qualified medical practitioner. RESULTS: The 50-59-year age group achieved the highest scores on the vasomotor and the depression scales in comparison to other age groups. Significant differences were also evident on the vasomotor and the depression scales on the basis of menopausal status, especially in perimenopausal women. Approximately 10% of women in the 60-79-year age group continued to experience vasomotor symptoms. CONCLUSION: Vasomotor symptoms, as assessed by the Greene Climacteric Scale, are common during the menopause transition and remain elevated for some years in a minority of older postmenopausal women. The norms presented in this study are appropriate for use in an Australian population. PMID- 15804733 TI - Effects of conjugated equine estrogens or raloxifene on lipid profile, coagulation and fibrinolysis factors in postmenopausal women. AB - OBJECTIVES: To compare the effect of conjugated equine estrogens (CEE) and raloxifene on lipid profile and hemostasis. MATERIALS AND METHODS: A double blind, randomized and parallel study was performed with 90 healthy postmenopausal women, aged 54 +/- 5 years, divided into three groups and submitted to daily therapy with either CEE 0.625 mg, raloxifene 60 mg or placebo for 4 months. The lipid profile, coagulation and fibrinolytic factors were analyzed. RESULTS: CEE increased the levels of high density lipoprotein cholesterol (HDL-C) from 49.0 to 56.8 mg/dl (p < 0.001), very low density lipoprotein cholesterol (VLDL-C) from 17.2 to 22.3 mg/dl (p < 0.001), and triglycerides from 86.0 to 111.7 mg/dl (p < 0.001), and decreased the levels of low density lipoprotein cholesterol (LDL-C) from 121.0 to 106.5 mg/dl (p < 0.001). The only significant effect of raloxifene was an increase in the levels of HDL-C from 46.0 to 47.8 mg/dl (p = 0.019). There was no significant reduction in LDL-C, from 115.5 to 110.2 mg/dl (p = 0.06), VLDL C, from 21.7 to 20.0 mg/dl (p = 0.201), and triglycerides, from 108 to 100 mg/dl (p = 0.201). CEE decreased the levels of fibrinogen, from 370.5 to 326.8 g/l (p = 0.039) and the levels of antithrombin III, from 99.5 to 93.2% (p < 0.001). Raloxifene decreased the levels of fibrinogen, from 354.7 to 302.0 g/l (p = 0.009) and the levels of antithrombin III, from 102.4 to 98.5% (p = 0.039). CEE increased levels of protein C from 103.7 to 115.3 mg/l (p < 0.001) and raloxifene did not change the levels of protein C (107.9 to 105.1 mg/l; p = 0.158). CEE decreased the antigen levels of tissue plasminogen activator (t-PA) from 8.8 to 6.8 U/ml (p < 0.001), and of plasminogen activator inhibitor (PAI-1) from 30.8 to 21.6 U/ml (p < 0.010), whereas raloxifene had no significant effect on either t PA, from 9.6 to 9.2 U/ml (p = 0.235) or PAI-1 antigen levels, from 32.1 to 30.4 U/ml (p = 0.538). CONCLUSION Both CEE and raloxifene exert significant effects on the lipid and coagulation profile. CEE had a more significant effect on fibrinolysis than raloxifene. These effects may have a significant impact on the cardiovascular risk that needs to be confirmed in larger studies. PMID- 15804734 TI - Vaginal rugae: measurement and significance. AB - OBJECTIVE: To devise a validated measure of vaginal rugae and assess the relationships between vaginal rugae and important clinical parameters. METHODS: Two techniques of assessing vaginal rugae were developed and their inter-/intra observer variability assessed. Examination variability was assessed using intraclass correlation and by way of an analysis of the absolute difference between the two rugal quantitations. After validating the assessment technique, the rugal quantitations of 88 women were compared to clinical parameters such as age, estrogen status, stage of prolapse, parity, history of anterior vaginal wall surgery, and body mass index. Linear regression analysis was used to assess the relationships between vaginal rugae score and these clinical parameters. RESULTS: The mean age and body mass index of the subjects were 56 years (standard deviation (SD) +/- 13.8 years) and 30.4 kg/m2 (SD +/- 7.5 kg/m2), respectively. The median parity was 2 (range 0-11). A history of anterior vaginal wall surgery was present in 29% of subjects and 46% were estrogen-deficient. Scores for the two techniques to quantitate vaginal rugae were normally distributed. Both techniques demonstrated satisfactory interexaminer reliability. Increasing age and deficient estrogen status were found to be independent predictors of less vaginal rugae. CONCLUSIONS: Vaginal rugae can be reliably quantitated. Loss of vaginal rugae is associated with estrogen deficiency and advancing age. PMID- 15804735 TI - Age at menarche, abnormal glucose tolerance and type 2 diabetes mellitus: The Rancho Bernardo Study. AB - OBJECTIVE: We examined the association of age at menarche with risk of abnormal glucose tolerance (AGT) and type 2 diabetes in postmenopausal women. METHODS: Participants were 997 women aged 50-92 years who had a clinic visit in 1984-87, when reproductive and diabetes history were recorded, and an oral glucose tolerance test administered. Information on age at menarche was collected in 1989. Diabetes status was based on WHO 1999 criteria, and AGT on having impaired fasting glucose (IFG) or impaired glucose tolerance (IGT). RESULTS: The average age was 69.5 years; 12.5% had diabetes and 29.8% had AGT. Menarche was < 12 years in 14.5%, 12-15 years in 78.9% and > or = 16 years in 6.6%. Age and body mass index (BMI) predicted AGT (p < 0.001, p = 0.003, respectively), while age, BMI and family history of diabetes predicted type 2 diabetes (p < 0.001, p < 0.001, p = 0.02, respectively). Polynomial logistic regression showed no association of menarche with AGT or type 2 diabetes risk. In linear regressions, however, older age at menarche was associated with lower fasting (p = 0.01) and post-challenge plasma glucose (p = 0.03). Covariates, especially BMI, weakened these associations, although women with menarche > or = 16 years still had lower post challenge (0.86 mmol/l, p = 0.05) and fasting glucose (0.26 mmol/l, p = 0.11) than those with menarche < 12 years. CONCLUSIONS: Age at menarche was not associated with AGT or type 2 diabetes risk. Late age at menarche was inversely associated with fasting and post-challenge glycemia. Obesity may mediate the observed associations. PMID- 15804736 TI - Endometrial and vaginal effects of low-dose estradiol delivered by vaginal ring or vaginal tablet. AB - AIMS: The major aims of the study were to compare the safety of a continuous low dose estradiol-releasing vaginal ring (ESTring) to that of a vaginal estradiol tablet (Vagifem) on the endometrium and the relief of subjective symptoms and signs of urogenital estrogen deficiency. Quality of life and acceptability of treatment delivery were also assessed. STUDY DESIGN: A prospective, randomized study in which women were assigned in a 2:1 ratio to ESTring and Vagifem and followed for 12 months. The primary endpoint was endometrial safety, based on the results of ultrasound measurement of endometrial thickness and a progestogen challenge test at baseline and week 48. Efficacy was determined by subjective assessment of urogenital estrogen deficiency symptoms at baseline and weeks 3, 12, 24, 36 and 48 and assessment of signs of vaginal epithelial atrophy by the clinician at baseline, 12 and 48 weeks. In addition, pelvic floor strength, vaginal cytological evaluation and pH, bacteruria and patient acceptability were assessed. Quality of life was assessed using a menopause-specific quality-of-life questionnaire and a 2-day bladder diary at baseline and 12 and 48 weeks. The comparability of the two groups was assessed using ANOVA, chi2 or Fisher's exact tests. RESULTS: A total of 126 women were randomized to ESTring and 59 to Vagifem. There was no statistical difference between the groups in the alleviation of symptoms and signs of urogenital estrogen deficiency. Maturation indices increased in both groups, from generally atrophic at baseline to proliferative or highly proliferative at 48 weeks. After 48 weeks of treatment, there was no statistically significant difference in endometrial thickness between the two groups. A statistically smaller proportion of bleeding/spotting occurred in the ESTring group (n = 0) compared to the Vagifem users (n = 4). Estradiol and total estrone serum levels increased during treatment in both groups but remained within the normal postmenopausal range. General health status in both groups was unchanged but the urogenital component of health burden was significantly improved in both groups. Bladder diary variables showed no differences between treatment groups. CONCLUSION: Equivalent endometrial safety and efficacy in the relief of the symptoms and signs of urogenital estrogen deficiency were demonstrated for the 12 months' use of a low-dose estradiol releasing vaginal ring and a vaginal estradiol tablet. PMID- 15804740 TI - Effectiveness of topical taurolidine versus ciprofloxacin, ofloxacin, and fortified cefazolin in a rabbit Staphylococcus aureus keratitis model. AB - PURPOSE: Taurolidine is a broad-spectrum, non antibiotic antimicrobial agent, not previously tested against the common causes of bacterial keratitis. This study, employing an experimental rabbit model of Staphylococcus aureus keratitis, investigated the effectiveness of topical taurolidine in reducing the number of bacteria, and its effectiveness was compared with topical ciprofloxacin, ofloxacin, and 5% cefazolin. METHODS: The right corneas of all rabbits were intrastromally injected with 100 colony-forming units of Staphylococcus aureus ATCC strain 25923. The animals were divided into the following seven groups: Group 1 (6 rabbits) received taurolidine, group 2 (6 rabbits) received ciprofloxacin, group 3 (6 rabbits) received ofloxacin, group 4 (6 rabbits)received cefazolin, group 5 (5 rabbits) received polyvinylpyrrolidone (vehicle),group 6 (4 rabbits) received sterile water, and group 7 (4 rabbits) was left un-treated (control group). The eyes were topically treated every 30 min with the above-mentioned substances from 4 to 9 h postinjection. One hour after the last drop administration (at 10 h postinjection), signs of inflammation were scored in a masked fashion by slit-lamp examination. Then, their corneas were processed. The number of colony-forming units (cfu) per cornea in all eyes was also determined. RESULTS: All antimicrobial (taurolidine, ciprofloxacin, ofloxacin, and cefazolin) treatments significantly reduced cfu numbers and slit lamp examination scores compared with untreated eyes, eyes that received the vehicle, or eyes with sterile water (all p values <0.05). Regarding cfu numbers, although taurolidine therapy was significantly less effective than ciprofloxacin or ofloxacin,there was no significant difference between taurolidine and cefazolin groups.However, taurolidine had similar clinical examination scores with the other antimicrobials, while it had lower scores than the vehicle, sterile water, or un-treated eyes. CONCLUSIONS: The results obtained in this study suggest that topicaltaurolidine is an effective, novel ocular chemotherapeutic agent for the therapy of rabbit experimental Staphylococcus aureus keratitis. This drug may be a useful and promising ocular antimicrobial. PMID- 15804742 TI - Ultrasonographic tissue characteristics of mushroom-shaped uveal melanoma. AB - PURPOSE: To evaluate the ultrasonographic tissue characteristics of mushroom shaped versus dome-shaped uveal melanoma and to compare them with the tumor vasculature. METHODS: We examined clinically and ultrasonically 18 dome-shaped and 11 mushroom-shaped uveal melanomas (29 consecutive patients; mean age 57 years) and compared their normalized A-scan patterns. Using histological sections from a different set of patients (12 dome- and 17 mushroom-shaped tumors), we calculated the percentage of area covered by blood vessels. RESULTS: A-scans of dome-shaped tumors presented low homogeneous reflectivity, whereas the heads of mushroom-shaped tumors had high and irregular reflectivity (p < 0.001). The reflectivity of their bases was low and similar to dome-shaped tumors. The blood vessel area in the histological sections was significantly larger in the heads of mushroom-shaped tumors (0.88 +/- 0.14%), compared both to their bases (0.29 i 0.06%) (p < 0.001) and to dome-shaped tumors (0.07 i 0.03%) (p < 0.0001). CONCLUSIONS: The ultrasonographic pattern suggests vascular congestion in the heads of mushroom-shaped tumors,probably due to strangulated blood vessels in the tumor neck. PMID- 15804741 TI - The effect of the menstrual cycle on the optic nerve head analysis of migrainous women. AB - PURPOSE: To determine the effect of the menstrual cycle on the optic nerve head topographic analysis of normally menstruating migrainous women. MATERIAL AND METHODS: Randomly selected one eye of 44 migrainous and 49 healthy control women with regular menstrual cycles were included in the study. All subjects underwent complete ocular examination. Optic nervehead topographic analysis were performed using a confocal scanning laser ophthalmoscope, HRT II (Heidelberg Retinal Tomograph II, software version 1.6;Heidelberg Engineering, Heidelberg, Germany). They were repeated for two times during the menstrual cycle: in follicular phase (7th to 10th day of the cycle) and in the luteal phase (days 3 to 4 before the menstrual bleeding). Serum estradiol, progesterone, and luteinizing hormone measurements were repeated at each menstrual phase. RESULTS: The mean age of migrainous and control subjects were 31.5 + 5.1 years and 33.4 +/- 3.7 years, respectively (P > 0.05). Their mean disc areas were 2.26 +/- 0.46 mm(2) and 1.95 +/- 0.39 mm(2), respectively(P < 0.05). Control subjects did not demonstrate any difference in the disc topography (P > 0.05). The parameter rim volume decreased, while the parameters cup volume and cup shape measure increased significantly in the luteal phase of the migrainous women (all P values <0.05). Mean intraocular pressure of the migrainous women decreased significantly in luteal phase (P < 0.05). CONCLUSION: Significant differences exist in the optic rim and cup parameters during the menstrual cycle of the migrainous women. Further clinical trials on ocular blood flow changes during the menstrual cycle of the migrainous women may highlight the role of sex steroids in the optic nerve head of the migrainous women. PMID- 15804743 TI - The effect of a beta-adrenoceptor antagonist on accommodative adaptation in Hong Kong children. AB - PURPOSE: Increased susceptibility to nearwork-induced accommodative adaptation has been suggested as a risk factor for myopia development. We investigated whether accommodative adaptation may explain in part the high prevalence of myopia in Hong Kong children and examined the effect of beta-antagonism with topical timolol maleate on accommodative adaptation. METHODS: Thirty children (10 emmetropes and 20 myopes) aged between 8 and 12 years were recruited. Tonic accommodation was measured before and after 5 min of video game-playing using an open-field Shin-Nippon autorefractor. Measurements were repeated 30 min after timolol instillation. RESULTS: Children with progressing myopia demonstrated accommodative adaptation following the near task, whereas stable myopes showed counter-adaptive, hyperopic accommodative changes. Timolol increased the magnitude of accommodative adaptation in the stable myopes but had little effect on responses of the progressing myopes or emmetropes. CONCLUSIONS: Neuropharmacological modulation of the accommodative system may have a possible etiological role in the progression of myopia. PMID- 15804744 TI - VIP immunoreactivity in human aqueous humor. AB - PURPOSE: To demonstrate the presence of vasoactive intestinal pep-tide (VIP) immunoreactive molecule in the human aqueous humor collected from eyes undergoing either cataract or glaucoma surgeries and to identify them asses of molecules responsible for the VIP immunoreactivity. METHODS: Aqueous humor specimens were collected by parencentesis from nine cataract patients and also from eight patients undergoing glaucoma surgery following the creation of the limbal based conjunctival flap, partial dissection of the scleral flap, and application of mitomycin-C. The aqueous humor specimens were analyzed by radioimmunoassay to determine the level of VIP immunoreactivity. Specimens from 10 other cataract patients were pooled and analyzed for VIP immunoreactivity by Western blot analysis. RESULTS: Levels of VIP immunoreactivity in aqueous humor of cataract and glaucoma patients were significantly different and were 610 +/- 160 and 260 +/- 64 pg VIP/ml, respectively (p = 0.03), while there was no correlation between the donor age and the level of VIP immunoreactivity. VIP immunoreactivity was detected as a single molecule with a molecular weight of 9000. CONCLUSIONS: The disease status and the treatments of the eye that led to surgery and procedures applied to the eye immediately before aqueous humor collection, but not the age of the patients, affected the level of VIP immunoreactivity in the aqueous humor. The relationship between the 9000 Da VIP-immunoreactive molecule and the authentic (3326 Da) VIP remains to be studied. PMID- 15804745 TI - CXCR2 Expression on neutrophils is upregulated during the relapsing phase of ocular Behcet disease. AB - PURPOSE: To search for markers of Behcet disease (BD) activity,we measured CXCR1 and CXCR2 levels on the circulating leukocytes of patients suffering from ocular BD. METHODS: Peripheral blood leukocytes were harvested from healthy volunteers (n = 16) and ocular BD patients (n = 35). The patients consisted of 15 individuals in relapsing phase (6 with prednisolone treatment) and 20 individuals in remission phase (9 with prednisolone treatment). Expression of CXC chemokine receptors (CXCRs) on leukocytes (including lymphocytes, monocytes, neutrophils) was measured using flow cytometry. RESULTS: Without prednisolone treatment, CXCR2 expression (on both total leukocytes and neutrophils) in relapsing phase was significantly higher than in remission-phase patients or normal individuals. By contrast, no significant difference was detected in the expression of CXCR1 between any ofthe groups. Importantly, low-dose prednisolone therapy reduced CXCR2 expression on neutrophils. CONCLUSIONS: CXCR2 has a potential role in promot-ing uveitis during ocular attack and might also be a useful marker for disease activity. PMID- 15804746 TI - Histamine induces opposing vasoactive effects at different levels of the ocular vasculature. AB - PURPOSE: To characterize the vasoactive properties and receptors of histamine in porcine retinal arterioles compared to that in the long posterior ciliary artery. METHODS: Isolated perfused porcine retinal arterioles were used to study vessel diameter in response to histamine (10(-10) to 10(-4) M). Ring segments from the long posterior ciliary artery were used to study contractile force in response to histamine. RESULTS: Histamine-induced vasodilatation in pig retinal arterioles but contraction in the long posterior ciliary artery. In retinal arterioles, the involvement of H(2) receptors was implied by the attenuation of the histamine response in the presence of cimetidine. Further attenuation of the histamine response by pyrilamine suggests the involvement of H1 receptors. In retinal arterioles denuded of endothelial cells, the histamine response was only slightly reduced. CONCLUSION: Histamine induces opposing vasoactive effects at different levels of the porcine ocular vasculature. PMID- 15804747 TI - Highly efficient ex vivo gene delivery into human corneal endothelial cells by recombinant adeno-associated virus. AB - PURPOSE: Gene delivery at high efficiency is crucial for cornea endothelial cell gene therapy. This study investigated the efficiency of gene transfer by recombinant adeno-associated virus (rAAV) in an organ culture system. METHODS: Human cornea tissue was exposed to rAAV delivering green fluorescent protein (ss rAAV2-CMV-GFP) for one hour and then cultured at 31 degrees C for 2 weeks in a medium supplemented with growth factors. Endothelial cells expressing GFP gene were then identified. RESULTS: High-efficiency gene transfer was found in over 90% of endothelial cells. Gene expression could be detected within 24 hours and remained stable up to 2 weeks in the organ culture system. CONCLUSIONS: The high delivery efficiency and rapid induction of gene expression indicate that rAAV is a promising vector for cornea endothelial cell gene therapy for ocular diseases. Organ culture at 31 degrees C using culture medium supplemented with growth factors significantly facilitates gene transfer into human corneal endothelium. PMID- 15804748 TI - Laser Doppler flowmetry in asymmetric glaucoma. AB - PURPOSE: To evaluate the relationship between extent of glaucoma damage and optic nerve blood flow, we investigated optic nerve head bloodflow in patients with asymmetric glaucoma damage between their two eyes and also in glaucomatous eyes with asymmetric damage inferiorly versus superiorly. METHODS: From our institutional practice, 16 subjects with asymmetric glaucoma damage between their two eyes and 25 eyes of 20 patients with asymmetric damage superiorly versus inferiorly were included in the study. Determinations of relative optic nerve head blood flow, velocity, and volume were obtained with laser Doppler flowmetry in the superotemporal and inferotemporal neuroretinalrim and in the cup. Means of flow (Flow(3)), velocity (Vel(3)), and volume (Vol(3))were calculated from these three measurement sites, and eyes or hemidisks with greater glaucomatous damage were compared to eyes or hemidisks with less damage. RESULTS: For subjects with asymmetric glaucoma damage between eyes, Flow(3) and Vel(3) were significantly lower in the eyes with worse glaucoma damage(mean difference = 2.09, p = 0.005, and mean difference = 0.05, p = 0.002,respectively). When comparing optic disks displaying within-eye asymmetry, the hemidisk with greater damage showed significantly lower blood velocity than the hemidisk with less damage (mean difference = 0.05, p = 0.013); however,no difference in blood flow or volume was detected. CONCLUSIONS: This study provides additional evidence that impaired optic nerve circulation is associated with the extent of glaucomatous pathology. PMID- 15804749 TI - Intraretinal pH in diabetic cats. AB - PURPOSE: To examine intraretinal extracellular H+ concentration([H+]o) in diabetic cats. METHODS: Double-barreled H+-selective microelectrodes were used to measure [H+]o as a function of retinal depth ([H+]o profiles) in four cats with different stages of diabetic retinopathy. Profiles from "normal"and "damaged" areas of the retina were compared to profiles previously obtained from healthy cats. RESULTS: In the healthy retina, [H+]o is generally highest in the middle of the retina and decreases toward the choroid and the vitreous. In 48 % of the profiles from diabetic animals with visible retinopathy, the inner retinal gradient was reversed so that the vitreous was more acidic than the middle of the retina. The profiles with reversed inner retinal gradients were classified as damaged. On the average, the inner retina tended to be 0.07-0.08 pH units more acidic in diabetic animals than in healthy normoglycemic animals, but of similar acidity to healthy hyperglycemic animals. In areas with damaged inner retinal gradients, net H+ production in the outer retina was also impaired. CONCLUSIONS: While the number of animals is small, we conclude that the [H+](O) distribution varied from normal to damaged in the same retina. Diabetes seems to lead to an acidification of the inner retina that appears to be at least partly related to hyperglycemia and which may be important in the progression of retinopathy. PMID- 15804750 TI - Perspectives on the genotoxic risk of styrene. AB - Styrene is a highly reactive monomer widely used in the plastics industry. The potential for styrene to produce genotoxic effects has been studied extensively in experimental systems. Styrene can induce sister chromatid exchanges (SCE) and chromosome aberrations (CA) in vitro under test conditions that enhance metabolism of styrene to styrene 7,8-oxide (SO)or reduce detoxification of 50 by epoxide hydrolase. The in vivo animal data indicate that styrene is not clastogenic at concentrations (doses) likely encountered by humans under ambient or occupational exposure conditions. DNA binding studies with styrene in rats and mice demonstrated no increased adducts in mice compared to rats or in mouse lung compared to liver. As a result, DNA adducts in the lungs are unlikely to be the sole explanation of the development of lung tumors in mice exposed to styrene for 2 yr. Some epidemiological studies reported that DNA and/or protein adducts and DNA strand breaks result from occupational exposure to styrene and/or 50. Results of some of these studies, how-ever, are difficult to interpret, given that the statistical significance of reported effects (SCE, CA, and micronucleus formation) was often near or at p values of .05; dose and/or temporal response relationships often were missing; confounding variables could not be excluded; and, concomitant exposures to other industrial chemicals that are potentially genotoxic may also have occurred. These studies suggest that styrene, through metabolism to SO, could be clastogenic in humans at workplace levels in excess of 125 mg/m3. However, results from controlled animal studies involving in vivo exposure to styrene alone do not show clastogenic effects at exposures of up to 1500 mg/m3/d. In any event, these studies show that there is an apparent threshold for styrene-mediated effects. PMID- 15804751 TI - Influence of functional group substitutions on the carcinogenicity of anthraquinone in rats and mice: analysis of long-term bioassays by the National Cancer Institute and the National Toxicology Program. AB - The carcinogenic activities of anthraquinone and six derivatives were compared and contrasted. Studies included representatives of amino, alkyl, nitro, hydroxy, or halogen-containing anthraquinones, with the purpose of uncovering general structure-activity relationships. Anthraquinone, 2-aminoanthraquinone, 1-amino-2 methylanthraquinone, 2-methyl-1-nitroanthraquinone,1-amino-2,4 dibromoanthraquinone, 1,4,5,8-tetraaminoanthraquinone, and 1,3,8-trihydroxy-6 methylanthraquinone (of varying purities) were administered via feed to Fischer 344/N rats and B6C3F, mice. In rats, anthraquinone induced tumors in the liver, kidney, and urinary bladder. A 2-amino substitution narrowed the carcinogenicity to the liver, while multiple amino substitutions led to a carcinogenic response in the urinary bladder alone. A methyl substitution ortho to a 1-aminogroup preserved the hepatic and renal neoplasms seen with the parent anthraquinone, but did not induce urinary bladder tumors; amino or bromo substitutions para to a 1 amino group were related to urinary bladder neoplasms. The intestine may have been a target organ for bromine-substituted anthraquinones. The presence of a nitro group altered the targets of carcinogenicity, and skin tumors may have been associated with this particular functional group in both rats and mice. Over-all for mice, the findings were somewhat different and limited by the small number of common target organs. The parent anthraquinone was clearly carcinogenic only to the liver. There were no other effects of single amino substitutions, in the presence or absence of an additional methyl group, on the carcinogenicity or the site of carcinogenesis of anthraquinone in mice. Multiple amino substitutions diminished, while bromine substitutions enhanced the carcinogenicity induced by anthraquinone and extended the target organs to include forestomach and lung. PMID- 15804752 TI - The effects of electromagnetic fields from power lines on avian reproductive biology and physiology: a review. AB - Electrical power lines are ubiquitous in the developed world and in urban areas of the developing world. All electrical currents, including those running through power lines, generate electric and magnetic fields (EMFs). Electrical power lines, towers,and distribution poles are used by birds for perching, hunting, and nesting. Therefore, many bird species, like humans, are exposed to EMFs throughout their lives. EMFs have been implicated in adversely affecting multiple facets of human health,including increasing the risks of life-threatening illnesses such as leukemia, brain cancer, amyotrophic lateral sclerosis, clinical depression, suicide, and Alzheimer's disease. A great deal of research and controversy exists as to whether or not exposure to EMFs affects the cellular, endocrine, immune, and reproductive systems of vertebrates. Laboratory work has used mice, rats, and chickens as models for this EMF research in an effort to understand better the possible implications of EMF exposure for humans. However, EMF exposure of wild birds may also provide insight into the impacts of EMFs on human health. This review focuses on research examining the effects of EMFs on birds; most studies indicate that EMF exposure of birds generally changes, but not always consistently in effect or in direction, their behavior, reproductive success, growth and development, physiology and endocrinology, and oxidative stress under EMF conditions. Some of this work has involved birds under aviary conditions, while other research has focused on free-ranging birds exposed to EMFs. Finally, a number of future research directions are discussed that may help to provide a better understanding of EMF effects on vertebrate health and conservation. PMID- 15804753 TI - Lesch-Nyhan Disease. AB - The first description of Lesch-Nyhan disease was in 1964; the first two patients were seen in 1963. The disease has caught the imagination of a variety of clinicians and scientists. The clinical picture is striking, combining spasticity, involuntary movements, and cognitive retardation with self-injurious behavior and the manifestations of gout. Biochemically, the overproduction of uric acid--the end product of purine metabolism--was, when measured, the largest ever seen. The disease is now well understood on a molecular basis. Enzyme analysis and mutational analysis have made available a full range of genetic testing, including diagnosis, carrier detection, and prenatal diagnosis. Therapy with allopurinol has been effective for those manifestations the disease shares with gout. Treatment for the neurological and behavioral features of the disease remains elusive. PMID- 15804754 TI - Medical societies and insanity in late-eighteenth-century London: the fight between Andrew Marshal and John Hunter. AB - In 1789, at a meeting of a small London society for the promotion of medical and surgical knowledge, Andrew Marshal (1742-1813) and John Hunter (1728-1793) engaged in heated debate regarding the association between mania and the structure of the brain. Marshal claimed to have observed abnormalities when dissecting brains of those who died insane and Hunter denied this connection. At the next meeting a scuffle between them ensued and they had to be parted. Although Marshal did not publish his observations during his lifetime, they were assembled by his assistant in 1815. Marshal's descriptions of the brains of hydrophobics and maniacs are worthy of note. PMID- 15804755 TI - NEUROwords Dr. Thomas Willis' famous eponym: the circle of Willis. PMID- 15804756 TI - Localization and William Macewen's early brain surgery Part II: The cases. AB - This paper contains Part II of an evaluation of the claims made for the priority of Macewen's pre-1884 brain surgery over that of Bennett and Godlee. Although the primary sources--Macewen's Private Journals and the Ward Records of the Glasgow Royal Infirmary--confirm that the operations were carried out when Macewen said they were, problems with the sources make it difficult to evaluate the extent to which he actually used knowledge of localization bearing on the claim for his priority evaluated. Part II concludes with citations from archival correspondence and the contemporary and near contemporary medical press confirming Macewen's priority. PMID- 15804757 TI - How the modern world began: Stephen Gaukroger's Descartes' system of natural philosophy. PMID- 15804760 TI - Letter to the editors: did a stone save Rome from destruction? PMID- 15804763 TI - Immunological factors in the pathogenesis and treatment of age-related macular degeneration. AB - Recent findings indicate that immunological factors are involved not only in the pathogenesis of age-related macular degeneration (AMD), but also in its treatment. Earlier data showing the presence of inflammatory cells in affected areas of AMD retinas support this statement. Although a possible role for autoimmunity was initially suggested, it has never reached general acceptance. Microorganisms have also been implied in the pathogenesis of AMD. Both serum antibacterial antibody levels and positive DNA tests from neovascular membranes have pointed to a possible role for Chlamydia pneumoniae in the pathogenesis of AMD. New data is providing evidence for the hypothesis that deposits between Bruch's membrane and the retinal pigment epithelium (RPE) cell layer may act as a stimulus for the local activation of the complement system. This may lead to a further growth of the deposits due to the strong chemotactic activity of certain complement activation products (such as C5a) with an influx of inflammatory cells. The buildup of cells and extracellular deposits may lead to local ischemia resulting in the activation of RPE cells. These activated RPE cells are thought to release angiogenic stimuli leading to choroidal neovascularization, which is the most serious complication of AMD. The fact that immunosuppressive drugs such as triamcinolone acetonide and anecortave acetate are capable of inhibiting choroidal neovascularization is consistent with an inflammatory component in the pathogenesis of AMD. Specific immunotherapy directed at certain cytokines or growth factors is now being investigated at both the animal and patient levels. Various clinical trials involving engineered antibodies are now being applied to block angiogenic factors such as the vascular endothelial growth factor (VEGF). An approach using gene therapy to influence angiogenesis by inducing the production of the pigment epithelium-derived factor (PEDF) was able to block neovascularization in an experimental murine model. Besides trying to block ongoing processes in AMD, retinal transplantation is now also being investigated as a treatment option. The fact that the retina is possibly an immunoprivileged tissue in combination with experimental data showing that the subretinal space is an immunoprivileged site is an indication that transplantation would not suffer from the rejection process. A larger obstacle is the question whether transplanted retinal tissue will regain its functional properties. PMID- 15804765 TI - Combined orbital floor betamethasone and depot methylprednisolone in uveitis. AB - PURPOSE: To evaluate the outcome of orbital floor injection of betamethasone and methylprednisolone in the management of uveitis, with particular reference to its efficacy in avoiding increase in systemic immunosuppressive drugs. METHODS: A sample of all patients attending the Uveitis Service at Moorfields Eye Hospital was carried out over a three-month period. All patients who had received one or more orbital floor injections of betamethasone (4 mg) and methylprednisolone (Depo-Medrone; 40 mg) in the management of their uveitis were identified and the case notes reviewed. RESULTS: A total of 48 patients who had received 75 orbital floor injections were identified during the sample period. Most patients had either panuveitis (n = 24) or pars planitis (n = 14). One-month post-injection visual acuity (VA) had improved by at least one Snellen line after 33/75 injections (44%) and at least two lines after 11/75 (15%), was unchanged after 25 (33%), but had deteriorated in 14 (19%). VA was not recorded after 3/75 injections. The only complications recorded were periorbital haemorrhage (n = 1) and complaints of persistent pain after an injection (n = 1). Symptoms (pain and subjective vision) improved after 36/75 injections (47%), were unchanged after 24 (32%), and were reported as worse after five (7%). Additionally, three patients reported an improvement in symptoms which lasted less than one month. Signs other than VA were improved after 28/75 (37%), were unchanged after 38 (51%), and deteriorated after only one injection. Of the 31 patients given orbital floor injections to avoid systemic therapy, seven (23%) subsequently went on to require it, and the clinical course then improved in 45%. CONCLUSIONS: Combined orbital floor injection of betamethasone and methylprednisolone can result in improved visual acuity, symptoms, and signs of inflammation in uveitis, and may therefore avoid the necessity for increased systemic medication. PMID- 15804764 TI - The immunomodulator vasoactive intestinal peptide (VIP) does not affect experimental autoimmune uveitis (EAU) in B10.RIII mice. AB - PURPOSE: Vasoactive intestinal peptide (VIP) exhibits immunomodulatory activities both in vivo and in vitro, including efficient inhibition of murine experimental arthritis. In this study, we investigated the effects of VIP treatment on the induction of experimental autoimmune uveoretinitis (EAU). METHODS: EAU was induced in B10.RIII mice by immunization with interphotoreceptor retinoid-binding protein (IRBP) using routine methods, but without treatment with pertussis toxin (PTX). VIP was injected i.p. at different doses into mice on alternate days. Mice were tested by conventional methods for ocular inflammation, antibody levels, lymphocyte proliferation, and cytokine release by cultured lymphocytes. RESULTS: Treatment with VIP, at different doses, had essentially no effect on the development of EAU or antibody production in the B10.RIII mice. The treatment did have variable effects on the low interferon-gamma production by lymphocytes of these mice. CONCLUSION: Unlike its inhibitory effect in the experimental arthritis system, VIP did not modulate the development of EAU in B10.RIII mice. PMID- 15804766 TI - Posterior intracorneal opacities in patients with HIV infection. AB - PURPOSE: To describe a keratopathy in patients with the human immunodeficiency virus (HIV) infection. METHODS: Retrospective review of ophthalmic and medical records, including laboratory evaluations, of seven patients with HIV infection and posterior intracorneal opacities. RESULTS: Each patient had a bilateral peripheral keratopathy located at the level of the Descemet membrane that was unassociated with intraocular inflammation. All patients were receiving highly active antiretroviral therapy (HAART). All patients had elevations in their CD4+ T-lymphocyte counts due to HAART prior to presenting with the corneal opacities. Five of the seven patients had elevated serum cholesterol, triglycerides, or both. Best-corrected visual acuity was 20/25 or better in six of the seven patients at the time of diagnosis, and vision remained stable through the follow up period in all patients (median follow-up: 25 months; range: 14-82 months). The corneal opacities remained unchanged in all seven over the follow-up period. CONCLUSION: These patients have a bilateral keratopathy that appears to be non progressive and has no effect on visual acuity. PMID- 15804767 TI - Visual outcome of juvenile rheumatoid arthritis-associated uveitis in adults. AB - PURPOSE: Juvenile rheumatoid arthritis (JRA) is the systemic disease most frequently associated in childhood uveitis. The disease may cause several ocular complications, visual impairment, and blindness. Recent studies revealed a more favorable ocular prognosis. Our purpose was to analyze the long-term visual outcome of JRA-associated uveitis. METHODS: Ocular complications and visual outcome in adult patients with JRA-associated uveitis were evaluated. Among 18 patients included in the study, uveitis was bilateral in 12 (66.7%) and unilateral in six (33.3%), for a total of 30 eyes with ocular involvement. RESULTS: The mean durations of JRA and its associated uveitis were 24.9 and 20.5 years, respectively. All eyes (100%) had at least one ocular complication. The most frequently observed ocular complications were cataract (83.3%), band keratopathy (60%), posterior synechia (46.7%), glaucoma (33.3%), hypotony (16.7%), and macular pathology (13.3%). Final visual acuity was impaired in 40% of the eyes, poor in 20%, and totally lost in 10%. Therefore, 70% of the eyes were either visually handicapped or totally blind. Most eyes underwent at least one surgical procedure. Inflammation was active at last examination in 63.3% of eyes. All patients were still treated topically and with systemic NSAID. Sixty one percent of the patients were using an immunosuppressive agent. CONCLUSION: JRA-associated uveitis still has a severe course and blinding potential. Patients suffer from uveitis and its complications even during the adulthood period. However, because our series represents a more severe subset of the disease, the outcome may be poorer than that of some other outcome studies. PMID- 15804768 TI - Effect of ketotifen fumarate, olopatadine, and levocabastine on ocular active anaphylaxis in the guinea pig and ocular immediate hypersensitivity in the albino rat. AB - PURPOSE: To study the effects of ketotifen fumarate, olopatadine, and levocabastine on ocular active anaphylaxis in guinea pigs and on ocular immediate hypersensitivity in albino rats. METHODS: Clinical grading scores and Evans blue dye leakage to eyelids and to eyeballs were assessed in five treatment groups (n = 10): ketotifen fumarate 0.025%, olopatadine 0.1%, levocabastine 0.05%, negative control, and positive control. RESULTS: At 20 minutes after challenge, edema scores for ketotifen-treated guinea pigs were statistically significantly lower than those for levocabastine or olopatadine. Active treatment significantly reduced vascular leakage in both models. Ketotifen significantly reduced vascular leakage in eyelids compared with the other drugs. In guinea pigs, vascular leakage in eyeballs was significantly reduced with ketotifen fumarate compared with olopatadine and levocabastine. CONCLUSIONS: In the guinea pig model, ketotifen was more effective than olopatadine and levocabastine at reducing conjunctival edema and vascular permeability in eyelids and eyeballs. In the rat model, ketotifen was more effective at reducing vascular permeability in eyelids than olopatadine and levocabastine. PMID- 15804769 TI - Causes of uveitis in a referral hospital in Ankara, Turkey. AB - PURPOSE: To evaluate the etiology of uveitis and its associated systemic findings in a referral hospital in Ankara, Turkey. MATERIALS AND METHODS: Clinical records of patients in our uveitis department were reviewed retrospectively and 419 eyes of 300 consecutive patients were included in the study. Cases of uveitis due to postoperative endophthalmitis, perforating injuries, and foreign bodies were excluded from the study. The data, including sex, age, clinical presentation, disease course and outcome, ocular involvement, etiology, and associated systemic findings, were entered into a computer database. Pearson chi-square statistical analysis was performed to analyze the data whenever necessary. RESULTS: The study subjects consisted of 162 males and 138 females, with a female to male ratio of 1:1.17 and an average age of 35.7 years. Anterior uveitis was the most common form of uveitis with 43.6%, followed by posterior uveitis (26.6%), panuveitis (20.6%), and intermediate uveitis (9%). Idiopathic uveitis (28.3%) was the most frequent cause of uveitis diagnosed, followed by Behcet's disease (26%) and rheumatic diseases (9%). In 215 (71.7%) out of 300 patients, a specific cause of uveitis with associated systemic findings was identified. Uveitis was attributed to an infectious etiology in only 41 patients (13.6%) of the entire study population. COMMENTS: The etiology of uveitis and its associated systemic findings may vary depending on the geographic distribution of patients and local factors. These results represent those of patients with uveitis referred to hospitals in central Anatolia. PMID- 15804770 TI - Indocyanine green and fundus fluorescein angiographic findings in patients with active ocular Behcet's disease. AB - PURPOSE: To define the indocyanine green (ICG) and fundus fluorescein angiographic (FFA) features of Behcet's disease. METHODS: The study included 49 eyes of 25 patients (19 males, 6 females; mean age: 34.1 years; age range: 14-68 years) with Behcet's disease and active ocular involvement that fulfilled the criteria of the International Study Group for Behcet's Disease. In each patient, FFA and ICG angiography were performed consecutively in the same session using a TRC-50I/A fundus camera (Topcon, Tokyo, Japan) coupled to an Image Net digitizing system. All images were analyzed and FFA, ICG, and clinical findings summarized. RESULTS: The mean duration of ocular involvement was 52.4 months (range: 2-240 months). FFA showed staining and dye leakage at the optic disc in 44 (89.8%) eyes and diffuse vasculitis in 36 (73.5%) eyes. Macular edema and ischemia were observed in 31 (63.3%) and 3 (6.1%) eyes, respectively. Eight (16.3%) eyes exhibited detectable retinal FFA alterations, but no abnormalities on ICG angiography. Disc ICG hyperfluorescence was observed in 23 (46.9%) eyes. ICG angiography revealed choroidal fuzziness in 16 (32.6%) eyes, hyperfluorescent spots in 13 (26.5%) eyes, and hypofluorescent plaques in 12 (24.5%) eyes. Eleven eyes (22.4%) showed no abnormal findings on ICG angiography. CONCLUSION: ICG and FFA complement each other as tools for diagnosing patients with Behcet's disease. FFA often serves as a fairly reliable guide. Although some ICG findings cannot be demonstrated by FFA, they are not specific or pathognomonic. We do not recommend performing both procedures for Behcet's disease. PMID- 15804771 TI - Interferon alpha-2a treatment for serpiginous choroiditis. AB - PURPOSE: To describe the management and long-term outcome of patients with serpiginous choroiditis (SPC) treated with interferon (IFN) alpha-2a. DESIGN: Prospective, non-comparative case series. PARTICIPANTS: Five patients (8 eyes) with active vision-threatening SPC, who failed to respond and/or presented with side effects to the combination of steroid-cyclosporin-A or chlorambucil, at a tertiary care referral center. INTERVENTION: The patients were treated with IFN alpha-2a (4,500,000 IU, 3 times a week for the first 3 months, followed by once a week for the next 3 months). Prednisone was also given initially, then tapered and discontinued. MAIN OUTCOME MEASURES: Visual acuity, control of inflammation, duration of drug-free disease remission, and side effects of IFN alpha-2a therapy. RESULTS: Argon laser photocoagulation (ALP) was applied in three eyes with extrafoveal choroidal neovascular membrane (CNM). All active lesions, including complicating subfoveal CNM in one eye, resolved within six months. The patients had no recurrences and recovered (4 out of 8; 50%) or maintained (4 out of 8; 50%) useful vision during the 16-48-month follow-up. Except for flu-like symptoms at the beginning of the therapy, no noticeable adverse reaction was observed. CONCLUSIONS: IFN alpha-2a seems to be a novel treatment option for SPC. In the five patients (8 eyes) with SPC, who were intolerant or unresponsive to the combination of steroid-cyclosporin-A or chlorambucil, IFN alpha-2a treatment prevented recurrences and stabilized the lesions during the long-term follow-up (16-48 months). These promising results support the use of this mode of therapy in controlled clinical trials. PMID- 15804772 TI - Deep intramuscular methylprednisolone treatment of recurrent scleritis. AB - PURPOSE: To evaluate the efficacy of deep intramuscular methylprednisolone (IMMP) in the treatment of recurrent scleritis. METHODS: A total of 15 patients with scleritis (12 anterior, 3 pan) underwent IMMP injection deep into the thigh. Thirteen patients were already being treated with oral non-steroidal anti inflammatory drugs, oral prednisolone, an oral immunosuppressive agent, or a combination of these drugs. A second IMMP injection was given to nine patients, making a total of 24 injections. Follow-up ranged from 4 to 18 months. RESULTS: There was a documented improvement in scleritis after 21/24 (87.3%) injections with a mean duration of improvement of 5.7 months. No patient required the introduction of oral corticosteroid or immunosuppressive agent, and only one patient required an increase in methotrexate to control the disease. No systemic, injection site, or ocular side effects were noted. CONCLUSIONS: Deep IMMP is a safe and effective treatment for scleritis. It ensures compliance, avoids the systemic side effects of oral corticosteroids, and is easily repeatable in the outpatient setting. PMID- 15804773 TI - Recurrent versus non-recurrent or no eye involvement in Behcet's disease. AB - PURPOSE: The aim of this study was to note the clinical features of patients with Behcet's disease who did not show recurrent episodes of eye involvement during long-term follow-up. METHODS: A retrospective review was made of 70 Japanese patients with Behcet's disease who were seen during a 23-year period from 1974 to 1996 and followed for five or more years at Okayama University Hospital. The 70 patients were divided into three groups: 36 patients with recurrent episodes of eye involvement; 13 patients with non-recurrent one-time only eye involvement; and 21 patients with no eye involvement at all during a follow-up period of five or more years. Clinical features were compared between these three groups. RESULTS: Female sex (p = 0.0263, chi-square test), young age at the onset (p = 0.0322, Kruskal-Wallis test), and genital ulceration (p = 0.0003, chi-square test) were significantly associated with the non-recurrent or no eye involvement patients. CONCLUSIONS: Female patients with onset at a young age and who show the three signs of skin lesions and oral and genital ulcerations may have non recurrent one-time only or no eye involvement at follow-up. A better visual prognosis can be expected for these patients. PMID- 15804774 TI - Endoretinal biopsy in establishing the diagnosis of uveitis: a clinicopathologic report of three cases. AB - PURPOSE: To report the value of endoretinal biopsy in establishing the diagnosis of severe posterior uveitis. METHODS: A clinicopathologic report of three patients with severe posterior uveitis unresponsive to corticosteroids and with a negative systemic workup. These patients subsequently underwent vitrectomy and endoretinal biopsy. RESULTS: Endoretinal biopsy provided an accurate diagnosis in the three cases, leading to appropriate treatment. Histopathologic examination of the three retinal samples confirmed the diagnosis of sarcoidosis, tuberculosis, and lymphoma. Targeted treatment improved the ocular conditions in all three cases. CONCLUSIONS: In selected cases of severe bilateral vision-threatening uveitis, retinal biopsy may be indicated as a last resort to confirm a suspected diagnosis of infectious disease or malignancy. PMID- 15804775 TI - Sympathetic ophthalmia after a hyphema due to nonpenetrating trauma. AB - PURPOSE: To report a case of sympathetic ophthalmia occurring after nonpenetrating ocular trauma. METHODS: Interventional case report. RESULTS: An 18 year-old white male presented with sympathetic ophthalmia four weeks after a hyphema from a nonpenetrating bungee cord injury. Vision was 20/20; there were keratic precipitates, anterior chamber cells, vitritis, and discrete yellow deep choroidal lesions consistent with Dalen-Fuch's nodules OU. The uveitis was treated with topical, oral, and periocular corticosteroids and the oral immunosuppressives cyclosporine and mycophenolate mofetil. Vision remained 20/20 OU for the first year of follow-up; vision OD then deteriorated to 20/50 over the next year with the formation of a posterior subcapsular cataract OD. CONCLUSION: Sympathetic ophthalmia may result from nonpenetrating ocular trauma. Ophthalmologists should be aware of this as prompt intervention is warranted. PMID- 15804776 TI - Ocular tuberculosis in acquired immune deficiency syndrome (AIDS). AB - AIM: To determine the prevalence and clinical features of ocular tuberculosis in patients with the acquired immune deficiency syndrome (AIDS). DESIGN: Prospective cross-sectional study. METHODS: Detailed history and ocular examination of 46 patients (92 eyes) in the outpatient department of an AIDS clinic. RESULTS: Seventeen of 46 (36.9%) patients had clinical and radiological evidence of pulmonary tuberculosis. Of these, four (23.5%) showed findings consistent with ocular tuberculosis. Lesions included tubercles (1 eye of 3 patients) and chorioretinitis (1 eye of 1 patient). Commonly, these patients had evidence of abdominal tuberculosis. Hemorrhages, cotton-wool spots, herpes zoster ophthalmicus (HZO), cytomegalovirus retinitis (CMVR), and disc edema were other non-tubercular AIDS-related lesions and were seen in seven of the remaining 42 patients (16.2%). CONCLUSION: Ocular tuberculosis was a common finding in this study and was found in profoundly immunocompromised patients with disseminated tuberculosis. Diagnosis of ocular tuberculosis may help reduce HIV/tuberculosis co-infection mortality. PMID- 15804777 TI - Vitrectomy and silicone oil tamponade as an initial surgery for retinal detachment after acute retinal necrosis syndrome. AB - PURPOSE: To show the feasibility of vitrectomy and silicone oil tamponade as an initial surgical procedure for retinal detachments with multiple tears that develop after the fulminant type of acute retinal necrosis syndrome. METHODS: In a prospective noncomparative interventional case series, vitrectomy, silicone oil tamponade, and encircling scleral buckling with a solid silicone, combined with phacoemulsification and intraocular lens implantation (except for one pseudophakic patient [Case 2]), were performed in three consecutive patients during the four-year period from 1999 to 2002. These patients had developed retinal detachment with multiple retinal tears after the resolution of acute retinal necrosis syndrome. As an initial standard treatment for acute retinal necrosis syndrome, all patients received maximum-dose intravenous acyclovir (1500 mg daily) for two weeks combined with intravenous prednisolone tapered from 200 mg daily. RESULTS: Case 1 was a 65-year-old woman who developed retinal detachment 44 days after the onset of acute retinal necrosis syndrome; Case 2, a 79-year-old man, developed retinal detachment 51 days after the onset; and Case 3, a 59-year-old man, developed retinal detachment 70 days after the onset. Before surgery, all patients showed multiple tears scattered on the detached atrophic retina extending over the entire midperipheral fundus with vitreoretinal adhesions and vitreous opacity. The retina was reattached during the initial surgery. The patients showed no recurrence of retinal detachment and maintained a visual acuity of 20/200 or 20/100 during the one- to three-year follow-up period. CONCLUSIONS: Vitrectomy and silicone oil tamponade could be a treatment option as the initial surgical procedure for retinal detachments that develop after the fulminant type of acute retinal necrosis syndrome. PMID- 15804778 TI - Lepromatous iridocyclitis. AB - A 36-year-old woman with no prior medical history presented with a unilateral decrease in vision and iridocyclitis to our hospital. External examination demonstrated multifocal, firm, elevated, non-tender skin nodules on the patient's face and left hand. In addition, slit-lamp biomicroscopy revealed gray, cheesy appearing nodules on the iris surface of the affected eye. Anterior chamber paracentesis and pars plana vitrectomy did not reveal further information. Skin nodule biopsy, however, demonstrated multiple fragmented organisms within histiocytes, consistent with leprosy. The iridocyclitis resolved after treatment with dapsone, corticosteroids, and rifampin and her vision returned to 20/20. PMID- 15804780 TI - Real time three-dimensional fetal echocardiography: is this really a paradigm shift? PMID- 15804781 TI - Plasma soluble vascular endothelial growth factor receptor-1 concentration is elevated prior to the clinical diagnosis of pre-eclampsia. AB - OBJECTIVE: Accumulating evidence suggests that the balance between vascular endothelial growth factor (VEGF), placental growth factor (PIGF), and their receptors is important for effective vasculogenesis, angiogenesis, and placental development. Recently, the soluble form of VEGFR-1 (sVEGFR-1), an antagonist to VEGF and PIGF, has been implicated in the pathophysiology of pre-eclampsia. Plasma sVEGFR-1 concentration is elevated in pre-eclampsia at the time of clinical diagnosis and correlates with the severity of the disease. The purpose of this study was to determine whether the concentrations of sVEGFR-1 in plasma of pre-eclamptic patients change prior to the clinical manifestations of the disease. METHODS: A longitudinal case-control study was conducted in normal pregnant women (n = 44) and patients with pre-eclampsia (n = 44). Blood sampling was performed at six intervals: (1) 7-16 weeks; (2) 16-24 weeks; (3) 24-28 weeks; (4)28-32 weeks; (5) 32-36 weeks; and (6) more than 37 weeks of gestation. To examine the relationship between plasmasVEGFR-1 concentration and interval to clinical diagnosis of pre-eclampsia, plasma samples of pre-eclamptic patients at different gestational ages were stratified according to the interval from blood sampling to clinical development of the disease into five groups: (1) at clinical manifestation; (2) 2-5 weeks; (3) 6-10 weeks; (4) 11-16 weeks; and (5) 17-25 weeks before clinical manifestations. Plasma concentrations of sVEGFR-1 were determined by enzyme-linked immunoassay. Parametric statistics and repeated measure procedures were used for the analysis. RESULTS: The mean plasma sVEGFR-1 concentration in pre-eclamptic patients before the clinical manifestation of the disease was significantly higher than in normal pregnant women at 24-28, 28-32, and 32-37 weeks of gestation (p = 0.02,p < 0.001, and p < 0.001, respectively). In contrast, no significant differences in the mean plasma sVEGFR-1 concentration between patients with pre-eclampsia and normal pregnant women were observed both at 7-16 weeks and 16-24 weeks of gestation (p= 0.1 and p= 0.9). Similarly, the mean plasma sVEGFR-1 concentration was significantly higher in pre-eclamptic patients than in normal pregnant women at clinical manifestation, at 2-5 weeks (mean 3.8 weeks), and at 6-10 weeks (mean 8.2 weeks) prior to the development of clinical pre-eclampsia (p < 0.001, p < 0.001, and p = 0.002,respectively). Among patients with early-onset pre-eclampsia (defined as gestational age of 34 weeks or less), the mean plasma sVEGFR-1 concentration was significantly higher in pre eclampsia (before clinical diagnosis) than in normal pregnant women at 24-28 (mean 26.4) weeks of gestation (p = 0.008). In contrast, among patients with the late-onset disease(defined as gestational age of more than 34 weeks), plasma sVEGFR-1 concentration in pre-clinical pre-eclampsia was significantly higher than in normal pregnant women at 28-32 (mean 30.2) weeks of gestation (p < 0.001). CONCLUSIONS: Plasma sVEGFR-1 concentration is elevated in pre-eclampsia prior to the clinical diagnosis of the disease. This elevation began 6-10 weeks prior to the clinical manifestations, and the increase was more pronounced at 2-5 weeks before the diagnosis, as well as at clinical presentation. Furthermore, in early-onset pre-eclampsia, plasma concentration ofsVEGFR-1 is elevated earlier than the late-onset disease. PMID- 15804782 TI - Trp64Arg polymorphism of the beta3-adrenergic receptor gene, pre-pregnancy obesity and risk of pre-eclampsia. AB - OBJECTIVE: Trp64Arg polymorphism of the beta3-adrenergic receptor (beta3-AR) gene was reported to be associated with increased risk of obesity and insulin resistance. Both traits have been implicated in the epidemiology and pathogenesis of pre-eclampsia. We investigated the relation of the beta3-AR gene to pre pregnancy obesity and risk of pre-eclampsia. The present study is a case-control study of 87 Caucasian pre-eclampsia cases and 214 controls. METHODS: Logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI), adjusted for maternal age and parity. Normotensive carriers of the variant allele (i.e. Arg64 allele) were 3.6 times more likely to be obese (body mass index (BMI) > or = 30 kg/m2) as compared with women not carrying the variant allele (OR = 3.6, 95% CI 0.8-15.0). RESULTS: Arg64 allele was more frequent in controls as compared with pre-eclampsia cases (11.7 vs. 5.3%, p < 0.01). After adjusting for age and parity, carriers experienced a 60% reduction in risk of pre eclampsia (OR = 0.4, 95% CI 0.2, 0.9) as compared with women not carrying this allele. The association between carriage of the variant allele and risk of pre eclampsia appeared to be modified by maternal BMI. Compared with lean carriers, lean women without the variant allele experienced only a small increase in the risk of pre-eclampsia (OR = 1.5; 95% CI: 0.6, 3.9). The OR for overweight women without the variant allele was 7.3 (95% CI 2.7-19.2). CONCLUSIONS: Our results suggest a puzzling relationship between the polymorphism and the risk of pre eclampsia. Future studies are needed to confirm these findings. PMID- 15804783 TI - The association between glucose challenge test, obesity and pregnancy outcome in 6390 non-diabetic women. AB - OBJECTIVE: To evaluate the association between obesity, glucose challenge test (GCT) and pregnancy outcome. METHODS: A prospective cohort study of 6854 consecutive gravid patients screened for gestational diabetes (GDM) using 50-gram GCT, at 24-28 weeks' gestation was performed. A screening value 130 mg/dl was followed by 100 gr oral GTT. Patients who were diagnosed with GDM were excluded. For purpose of analysis patients were categorized by prepregnancy BMI and by different GCT thresholds. Maternal outcome was defined by rate of preeclampsia, gestational age at delivery, cesarean section (CS) rate and the need for labor induction. Neonatal outcome was defined by fetal size (macrosomia/LGA), arterial cord pH, respiratory complications and neonatal intensive care unit (NICU) admission. RESULTS: Overall, a positive GCT result (GCT > or = 130 mg/dl) was identified in 2541/6854 (37%) women. GDM was further diagnosed in 464/6854 (6.8%) of subjects. In both groups of screening results ( > 130 mg/dl and < 130 mg/dl), the obese women were significantly older, gained more weight during pregnancy and had a lower rate of nulliparity in comparison to the non obese women. The obese women had higher rates of macrosomia, LGA and induction of labor. No difference was found in mean birth weight, the total rate of cesarean section, preterm delivery, 5 minute Apgar score < or = 7, mean arterial cord pH, NICU admission and a need for respiratory support in comparison to non obese women in both groups of screening results. A gradual increase in the rate of macrosomia, LGA and cesarean section was identified in both obese and non-obese women in relation to increasing GCT severity categories. CONCLUSION: Fetal size and cesarean section rate are associated with the degree of carbohydrate intolerance (screening results). Furthermore, obesity remains the main contributor impacting fetal size. PMID- 15804784 TI - Obstetric and neonatal outcomes associated with maternal hypothyroid disease. AB - OBJECTIVE: We sought to determine whether women with treated hypothyroid disease were more likely than women without thyroid disease to suffer adverse obstetric or neonatal outcomes or to deliver a child with a congenital anomaly. METHODS: Using an institutional database, we identified women with treated hypothyroid disease (n = 482) who delivered a baby at our institution during a 33-month period. We compared the occurrence of adverse obstetric or neonatal outcomes among these women to the occurrence among women without thyroid disease (n = 19,487). RESULTS: Women with treated hypothyroid disease were not at increased risk for delivering a baby with low birth- weight,fetal demise, or congenital anomaly compared to the control group. Women with treated hypothyroid disease were more likely to have chronic hypertension (2.3% vs. 1.2%, p = 0.03) and had an increased risk of pre-eclampsia (4.3% vs. 2.6%,p= 0.03) compared to women without thyroid disease. CONCLUSION: Women with treated hypothyroid disease are not at higher risk than the general population for adverse neonatal outcomes, but may be at increased risk for pre-eclampsia. PMID- 15804785 TI - The safety of low molecular weight heparin therapy during labor. AB - OBJECTIVE: Current recommendations are to discontinue low molecular weight heparin (LMWH) at least 24 hours prior to labor induction or administering epidural anesthesia. We assessed the safety of discontinuing LMWH 12-24 hours before delivery. METHODS: We evaluated the prevalence of hemorrhagic complications during labor, cesarean or epidural catheter placement in 284 women treated with enoxaparin during pregnancy as compared with 16132 untreated women. Treated participants were divided into subgroups by the various intervals between last LMWH dose hemorrhage-prone events (vaginal delivery,epidural, cesarean etc.). The rate of hemorrhagic complications and hemoglobin values were compared between the study and control groups. RESULTS: Postpartum hemorrhage was uncommon and occurred in 2.1% and 1.9% in study and control groups, respectively (p=0.13). Antenatal as well as postnatal hemoglobin values were very similar for treated and untreated women. No differences were noted between women who discontinued enoxaprin 12-24 hours before labor and those who discontinued treatment later with regard to maternal hemorrhagic complications. No spinal hematomas were report among 12792 treated and un-treated women who had epidural or spinal block. No hemorrhagic neonatal complications were encountered. CONCLUSION: Discontinuing LMWH more than 12 hours before delivery is safe in relation to maternal hemorrhagic complications. PMID- 15804786 TI - Changes in fetal presentation in twin pregnancies. AB - OBJECTIVE: The aim of the study was to describe the occurrence of spontaneous version in twins in the third trimester, as well as the correlation between antepartum and birth presentation. METHODS: Twin pregnancies were divided into four gestational age intervals: 20-23; 24-27; 28-31; and 32-36 weeks. Fetal presentation was categorized as cephalic (C) or non-cephalic (NC). Hospital records were reviewed to determine demographic factors, including mode of conception. Chi-square was used to compare distributions of presentations, and Cramer's V measure of association was used to correlate presentations in individual pregnancies between antepartum intervals and birth. RESULTS: A total of 207 pregnancies were included. The distribution of fetal presentations changed significantly through gestational age intervals (p < 0.001), although they were similar between 32-36 weeks and birth (p = 0.75). Correlation between antepartum and birth presentation in individual pregnancies strengthened throughout the four intervals. No correlation was seen between parity, gender, birthweight, or in vitro fertilization and fetal presentation or rate of spontaneous version. CONCLUSION: Many twins undergo spontaneous version in the third trimester, though there is excellent correlation between presentation at 32-36 weeks and birth. A cephalic presenting twin at > or = 28 weeks is highly likely to be in cephalic presentation at delivery. PMID- 15804787 TI - Cesarean delivery during second-stage labor: characteristics and diagnostic accuracy. AB - OBJECTIVE: To characterize dysfunctional labors that lead to cesarean delivery in the second stage and to assess the accuracy of diagnoses of abnormal fetal descent. METHODS: Thirty-one patients delivered by cesarean during the second stage because of abnormal labor or presumed cephalopelvic disproportion were studied and compared to 62 control cesarean cases delivered for the same indications in the first stage. The clinical diagnosis of dysfunctional labor that led to the cesarean was compared to the diagnosis made by retrospective analysis of the labor curves. RESULTS: Cases did not differ from controls delivered in the first stage in maternal age, race, parity, gestational age, weight gain, or the frequency of associated medical complications. The newborns were not significantly different in birth weight,ponderal index, sex, or the incidence of low Apgar scores. Among study patients, 94% had a second stage labor dysfunction determined by graphic labor analysis, predominantly arrest of descent (69%) and failure of descent (28%). In 79% of cases a dysfunctional first stage preceded the abnormal second stage. Among these first stage labor abnormalities, 68% were not recognized during the labor. CONCLUSION: Characteristics of patients delivered by cesarean during the second stage were similar to those delivered before full cervical dilatation. Second stage labor abnormalities were usually preceded by an abnormal first stage. There was considerable inaccuracy in the diagnosis of second stage labor dysfunction. PMID- 15804788 TI - Clinical pharmacokinetics of lyophilized recombinant human erythropoietin-alpha following single-dose subcutaneous administration in premature newborns. AB - OBJECTIVE: The aim of this study was to determine pharmacokinetic parameters after subcutaneous administration of a single dose (400 IU/kg) of lyophilized recombinant human erythropoietin-alpha (rhEPOalpha) to preterm newborns. The parameters determined were: maximum concentration (C(max)), time to reach maximum concentration (T(max)), absorption half-life (t(l/2abs)), volume of distribution (Vd), elimination half-life (t(l/2el)), clearance (C(L)), constant of elimination (k(el)) and area under the 0-72 h curve (AUC(0-72). METHODS: The study group comprised 20 premature newborns (eight males and 12 females) delivered in the Teaching Hospital, University of Sao Paulo. The inclusion criteria were birth weight < 1500 g or gestational age < or = 34 weeks, and clinical and hemodynamic stability. Serum erythropoietin (EPO) concentration was determined before and 1, 4, 6, 12, 24,48 and 72 h after subcutaneous administration of 400 IU/kg rhEPOalpha, and the pharmacokinetic parameters were calculated. RESULTS: There was a significant difference in serum EPO concentration between t72 and t0 (p = 0.001). Mean values (range) of the pharmacokinetic parameters were as follows: C(max), 739.8 (188.0-1390.0) mIU/ml; T(max), 7.7 (4.0-12.0) h; t(l/2abs), 2.9 (0.8-4.8) h, V(d), 0.705 (0.23-1.73) 1/kg; t(l/2e1), 14.9 (8.7-36.1) h; C(L), 0.032 (0.014-0.066) 1/h; k(el), 0.0475 (0.0200-0.0700); and AUC(0-72), 19058.2 (7648.0-34701.5) mIU/ml per h. The Spearman test showed no correlation between the pharmacokinetic parameters analyzed and the characteristics of the population studied. CONCLUSIONS: Studies evaluating the effectiveness of therapy with recombinant human erythropoietin in premature newborns have used various doses, administered at intervals between 24 and 48 h. The kinetics of absorption measured in our study supports the use of 400 IU/kg within an interval of no less than 72 h, together with therapeutic control of the drug and evaluation of the erythropoietic response. PMID- 15804789 TI - A study of neonatal swimming (water therapy) applied in clinical obstetrics. AB - OBJECTIVE: To study the significance of some clinical parameters related to neonatal 'swimming' (water therapy) during hospitalization. METHODS: Normal newborns were randomly divided into two groups to observe their birth weight, weight before discharge,time of first defecation and meconium turning yellow. Group one was the swimming (study) group, comprising a total of 223 newborns including 127 babies delivered after spontaneous vaginal delivery and 96 babies after Cesarean section. Group two was the bathing (control) group, comprising 154 newborns including 109 babies delivered after spontaneous vaginal delivery and 45 babies after Cesarean section. RESULTS: There was no significant difference in birth weight between the two groups (p > 0.05). However, the mean weight before discharge of the babies in the study group was 3.29 + 0.35 and 3.51 + 0.40 kg, spontaneous vaginal delivery vs. Cesarean section, compared with 3.09 + 0.38 and 3.17 + 0.48 kg, respectively, in the control group (p < 0.01). The corresponding mean times of meconium turning yellow were 39.15 + 15.88 and 39.02 + 13.60 h in the study group compared with 48.01 + 19.42 and 55.67 + 25.05 h in the control group. This difference was significant (p < 0.01), as was the difference between the time of first defecation (p < 0.05). CONCLUSION: Neonatal swimming can accelerate babies' growth in the early stage. PMID- 15804790 TI - Gestational diabetes exhibits lack of carnitine deficiency despite relatively low carnitine levels and alterations in ketogenesis. AB - OBJECTIVE: Previous studies have underlined the importance of the carnitine shuttle system and its dysfunction both in normal pregnancy and in type 1 and 2 diabetes. The objective of this paper was to delineate more systematically the role of the carnitine shuttle system in normal pregnancy and in gestational diabetes. METHODS: A total of 119 women matched for age comprised three groups: 40 normal adult non-pregnant women (NNP), 46 normal pregnant women with uncomplicated pregnancy (NP) and 33 women with gestational diabetes (GDM). The latter group was further subdivided into those being managed either by diet alone (25 women, GDM-D) or by insulin (8 women,GDM-I). The following biochemical parameters were assayed: fasting plasma total, free and acyl-carnitine, FFA and beta-OH-butyrate, together with several essential anthropometric parameters. RESULTS: Women with GDM, in contrast to the control groups, displayed the biochemical features characteristic of insulin resistance: higher body weight, higher BMI, higher skinfold and higher HbAlc levels. No differences on any parameters were found between the two GDM subgroups. Both NP and GDM groups had low levels of total carnitine compared to NNP control group, but surprisingly, the GDM group did not exhibit any further decrease of carnitine levels, as would have been expected by the combination of pregnancy and diabetes. Both groups, despite these low carnitine levels, had no clinical symptoms of carnitine deficiency. Furthermore, the GDM group displayed higher levels of FFA and beta hydroxybutyrate, which were statistically significant compared to the other two control groups. CONCLUSIONS: The data corroborate the negative effect of normal gestation on the carnitine shuttle system, while they document for the first time that GDM does not further affect the efficiency of the carnitine system. The mild effect of GDMon carnitine status could be explained by the concurrent increased gluconeogenesis, a process which does not affect directly carnitine metabolism. PMID- 15804791 TI - Prevention of necrotising enterocolitis: year 2004 and beyond. AB - The incidence of necrotising enterocolitis (NEC) has not changed significantly despite the dramatic advances in perinatal-neonatal care. The absolute number of survivors of NEC is expected to rise, as prevention of prematurity, the single most important risk factor for the illness, continues to be difficult. Prevention of NEC has thus become an area of research priority. Given the role of inflammatory mediators in its pathogenesis newer immune modulators are being studied as potential agents for prevention/treatment of NEC. Caution, however, is warranted because the failure of sepsis trials in adults has clearly indicated that the concept of down-regulating the inflammatory response is deceptively simple. Clinical trials of any such promising preventative agent(s) need to be designed carefully and must include long-term neurodevelopmental outcomes as almost an entire population of high-risk preterm neonates will be exposed to their adverse effects. As new frontiers continue to be explored, the proven benefits of simple and safe interventions like antenatal glucocorticoid therapy and the preferential use of breast milk for feeding high-risk neonates must not be forgotten. Given that a single effective agent is unlikely in the near future, utilizing a package of "potentially better practices" seems to be the most appropriate strategy to prevent and minimize NEC. PMID- 15804792 TI - Glioblastoma multiforme in pregnancy. AB - Brain tumors in pregnancy are uncommon occurrences. A 48-year-old female at 36 2/7 weeks of gestation presented with right-sided weakness and parasthesias, a facial droop, and an unsteady gait. Brain imaging studies revealed a left-sided parietal lesion impinging upon the lateral ventricle. She underwent a cesarean delivery and then a craniotomy with resection of a glioblastoma multiforme two days later. Despite treatment with radiation and chemotherapy, she died nine months later. Glioblastoma multiforme is a rare diagnosis during pregnancy which may present as a neurologic emergency. PMID- 15804793 TI - How much and for how long does the neonatal myocardium suffer from mild perinatal asphyxia? AB - Cardiac troponins can be useful in monitoring cardiac injury following perinatal distress. We report here an increase of cardiac troponin I (cTnI) to 2.84 microg/l at 3 weeks (age-related median: 0.07 microg/l) followed by normalization in a newborn with an uneventful clinical course after resuscitation at birth. Serial echocardiographs showed normal cardiac function. Such a time course of cTnI, not previously reported, could be due to either a greater sensitivity of biochemical markers than of instrumental tools or birth asphyxia. Larger studies are needed PMID- 15804795 TI - Inequality in access to CABG and PCI. PMID- 15804796 TI - Informed consent in routine healthcare. PMID- 15804797 TI - Factors influencing late allograft valve failure. AB - Allograft valves are a valuable valve replacement substitute in the surgical management of heart valve disease. It remains the valve substitute of choice in the reconstruction of the right ventricular outflow tract in children with congenital heart disease and in the Ross procedure. However, its durability remains suboptimal, particularly in children. This article reviews the mechanisms and factors implicated in late allograft dysfunction, with a focus on the evidence for an immunological cause for allograft valve failure. Unravelling the mechanisms of allograft valve failure may allow modification of the allograft to improve its long-term durability. PMID- 15804798 TI - Inequality in access to coronary revascularization in Sweden. AB - OBJECTIVES: To investigate social and gender equality in access to coronary revascularization for those treated for coronary heart disease in Sweden between 1991 and 2000. DESIGN: All Swedish residents between 25 and 74 years old with a hospital stay for coronary heart disease were eligible for the study, in total about 153,000 persons. The Swedish Hospital Discharge Register from 1988 through 2000 was used to define the study population. Poisson regression analyses were used to estimate the effect of socio-economic status on the likelihood for coronary artery bypass grafting (CABG) within 2 years. In the analysis of gender differences, the likelihood for percutaneous coronary intervention (PCI) was also included. RESULTS: Males were 1.5 times more likely to undergo revascularization procedures than females even after adjusting for confounding factors and the fact that women are less eligible for interventions. The analyses also showed significant socio-economic inequalities in access to CABG among men, but not among women. CONCLUSIONS: There are gender and socio-economic inequalities in access to cardiac procedures in Sweden. PMID- 15804799 TI - Trends in fatal and non-fatal coronary heart disease events in Finland during 1991-2001. AB - OBJECTIVE: To analyse the trends in fatal and non-fatal coronary heart disease (CHD) events in Finland during an 11-year period 1991-2001. DESIGN: Data on hospitalizations due to CHD in the Hospital Discharge Register were linked to the National Causes of Death Register in order to produce a Cardiovascular Disease Register including data on 271,771 events in 234,244 individuals. RESULTS: The annual average decline in the age-standardized CHD mortality rate was 5.2% (95% CI, -5.6, -4.8%) among men and 6.1% (-6.6, -5.6%) among women. The incidence of first myocardial infarction declined annually on average by 5.5% (-5.9, -5.1%) from 1991 to 1997 and by 2.4% (-3.0, -1.7%) from 1998 to 2001 among men. The respective changes among women were -5.9% (-6.5, -5.2%) and -1.7% (-2.7, -0.6%). The number of hospitalizations due to unstable angina pectoris increased between 1991 and 1996 (p = 0.0002) and remained stable for the rest of the study period. CONCLUSIONS: The Cardiovascular Disease Register is a powerful tool for epidemiological monitoring of cardiovascular diseases in Finland. PMID- 15804800 TI - Serum iron, infection and inflammation; effects on coronary risk. AB - OBJECTIVES: To explore the potential pathways of association between serum iron and coronary heart disease, with major emphasis on factors related to infections and inflammation. DESIGN: A nested case-control study with 215 cases (myocardial infarction or coronary death) and 215 matched controls over 8.5 years. Logistic regression analyses were used to compare relative risks in various serum iron high sensitive CRP-total leucocyte count-herpes simplex virus-1 antibody categories. RESULTS: Participants with low iron (< 17 micromol/l) had increased coronary risk with Odds Ratio (OR) of 2.1 (95% CI 1.1-3.8). Simultaneous elevation of hs-CRP and leucocyte count increased the risk substantially in those with low iron, OR 9.8 (95% CI 3.9-24.4). A combination of high herpes simplex virus-1 antibody level and low iron increased the risk modestly (OR 1.2), but when hs-CRP level was high simultaneously, the OR was 13.1 (95% CI 2.9-60.1). CONCLUSIONS: Our data suggest an association between low serum iron level and coronary risk. The association is not independent, but is related to the fact that chronic infections and inflammation are accompanied with low serum iron. PMID- 15804801 TI - Coronary aneurysm after implantation of a paclitaxel-eluting stent. AB - Formation of coronary aneurysm is a rare complication of stenting with bare metal stents, but based on experimental studies drug-eluting stents may induce toxic effects on the vessel wall with incomplete stent apposition, aneurysm formation and with the potential of stent thrombosis or vessel rupture. We present a 43 year-old man who developed a coronary aneurysm in the right coronary artery 6 months after receiving a paclitaxel-eluting stent. The patient was asymptomatic and the aneurysm was detected in a routine control. Angiography and intracoronary ultrasound demonstrated lack of contact between stent and vessel wall in a 15-mm long segment with maximal aneurysm diameter of 6.0 mm. The patient was successfully treated with a graft stent. PMID- 15804803 TI - Congenital heart defects -- occurrence, surgery and prognosis in a Danish County. AB - OBJECTIVE: To present population-based data on prevalence, surgery and mortality for infants and children up to 5 years of age with congenital heart disease (CHD). DESIGN: Data from the EUROCAT Registry of Congenital Malformations for Funen County, Denmark, 1986-1998. RESULTS: Five hundred and seventy-three infants and children were diagnosed with a CHD and livebirth prevalence was 7.9 per 1000 births. Thirty-two percent of all infants and children had an intervention (surgery or catheter treatment) performed. Eighteen percent died within the first 5 years with the majority of deaths within the first years of life. For 74% of all deaths, surgery had not been performed. There was a decline in mortality for 1994-1998 compared to 1986-1993 both as a percentage of all cases (p < 0.05) and all deaths per 1000 births (p = 0.13), and deaths within the first 28 d after surgery almost disappeared during the study period. CONCLUSION: Mortality and morbidity for infants and children with CHD is rather high although surgical mortality has improved considerably. Survival may be improved further for the small group of severely ill newborns dying before surgery. In newborns with multiple malformations, however, survival might not be possible or desirable. PMID- 15804802 TI - Unstable angina, stroke, myocardial infarction and death in aspirin non responders. A prospective, randomized trial. The ASCET (ASpirin non responsiveness and Clopidogrel Endpoint Trial) design. AB - BACKGROUND: Aspirin is widely used as an antiplatelet drug in patients with coronary heart disease. Despite documented clinical benefit, many patients on aspirin still experience severe cardiovascular events. Several laboratory reports have shown lack of platelet inhibition in 5-40% of aspirin-treated patients, and the term aspirin resistance has been introduced. The clinical relevance of these laboratory findings is, however, still unknown. New antiplatelet drugs have been developed, and the adenosin diphosphate (ADP) receptor inhibitor clopidogrel has at least the same efficacy as aspirin with an acceptable safety profile. Laboratory methods for determination of platelet reactivity and treatment efficacy have been complicated and time consuming. New methodologies, like the PFA-100 system, have made such analyses more suitable for clinical use. DESIGN: In the ASCET study, 1000 patients with documented coronary heart disease will be randomized to either continued treatment with aspirin 160 mg/d or change to clopidogrel 75 mg/d after initial determination of their platelet reactivity while on aspirin treatment. Clinical endpoints will be recorded for at least 2 years and related to the initial aspirin response. PMID- 15804805 TI - Chest wall motion and pulmonary function are more diminished following cardiac surgery when the internal mammary artery retractor is used. AB - OBJECTIVES: Pulmonary complications following cardiac surgery through sternotomy have been widely studied. The duration of these complications, however, has been less studied and the proposed alterations in chest wall mechanism even less. The purpose of this study was to investigate changes in chest wall motion and pulmonary function of cardiac surgery patients, where both the median and the internal mammary artery retractor was used (IMA group) and cardiac surgery patients, where only the median retractor was used (Median group). DESIGN: Subjects were 20 cardiac surgery patients with mean age 65 years (12 in the IMA group and 8 in the Median group). Bilateral respiratory movements (RMs) using the Respiratory Movement Measuring Instrument, lung volumes including vital capacity (VC), forced vital capacity (FVC) and forced expiratory volume (FEV1) using the Vitalograph Alpha were measured and pulmonary radiographs analyzed before, 3 and 12 months after the operation. ANALYSIS: Descriptive statistics, t-tests, Mann Whitney and Wilcoxon Signed Rank tests were used for analyses, p < or = 0.05. RESULTS: RMs were symmetrical in both groups prior to the operation and the differences in RMs and lung volumes between the groups were not significant. Three and 12 months postoperatively bilateral abdominal respiratory movements (ARM) were significantly less in the IMA group than in the Median group. Average left ARM were significantly less than the average right ARM in the IMA group 3 months postoperatively, while symmetrical in the Median group. Average FVC and FEV1 were significantly less in the IMA group than in the Median group 3 months postoperatively and FVC was still significantly less in the IMA group 12 months after the operation. CONCLUSION: The significantly more reduced ARM and lung volumes 3 months postoperatively in the IMA group than in the Median group suggests that the IMA retractor causes greater injury to the rib cage and the diaphragm. PMID- 15804804 TI - Patients' views of the consent process for adult cardiac surgery: questionnaire survey. AB - OBJECTIVE: Consent for surgical procedures has assumed increasing importance in surgical practice in recent days especially following the public inquiry into paediatric cardiac surgery deaths at Bristol in the UK. This study examines patient perceptions and recollections following surgical consent as currently practised in a UK cardiac unit. METHODS: One hundred consecutive patients who underwent cardiac surgery in a London teaching hospital from January to February 2003 were studied. Patients completed questionnaires a day before their discharge from the hospital. RESULTS: The majority of patients (89/100) responded that the information given at consent had been adequate or more than adequate. The time spent on the consent process was thought to be adequate by 91 patients. Eleven patients felt the consent had been insensitive. Several patients (38/100) felt use of booklets in preference to verbal explanations would be less intimidating. For most patients (94/100) the operation and postoperative course met their expectations; although 12 patients experienced untold complications, only five felt that they should have been informed of the possibility of the complication. Although most patients were informed of the risk of death during consent, at time of discharge 43 had forgotten the figure that had been quoted. Regarding the influence of media and publicity, 19 patients said that media had influenced their expectations of the consent process, 59 would have liked to see hospital league tables while 26 would have liked to know the mortality figures for their surgeon prior to giving consent. CONCLUSIONS: Our study shows that patients undergoing cardiac surgery are largely satisfied with our improved consent procedures in the post-Bristol era. Use of booklets may be a useful adjunct to verbal consent as currently practised. PMID- 15804806 TI - Surgical closure of patent foramen ovale in patients with suspected paradoxical embolism: long-term results. AB - OBJECTIVE: It is thought that a patent foramen ovale (PFO) is the crucial mechanism in patients with suspected paradoxical embolism and cryptogenic stroke. It has been hypothesized that closure of the PFO would prevent further cerebrovascular incidents. We describe our early and late experience with surgical closure of the PFO in patients with paradoxical embolism. PATIENTS AND METHODS: Between May 1994 and December 2001, 33 patients (26 men, 7 women; mean age, 55.2 +/- 8.7 years; range, 37-74) underwent surgical closure of a PFO at our institution. All patients had preoperatively suffered from a stroke and/or a transient ischemic attack, after which echocardiography showed a PFO. Mean follow up at 99 +/- 30 months (range, 10-111 months) was 100% complete. RESULTS: All patients survived the operative procedure. Early complications occurred in four patients (12%). Actuarial survival at 1, 5 and 8 years was 97 +/- 3%, 97 +/- 5% and 94 +/- 8%, respectively. At long-term follow-up all but two patients were alive. The deaths of these two patients were related to malignancy and ischemic heart disease, respectively. Two patients (6%) had suffered a residual cerebrovascular event after successful surgery. CONCLUSION: Surgical closure of PFO in patients with paradoxical embolism can safely be performed with a low risk of early mortality. Residual thromboembolic events were rare and in those few it occurred it did so with the interatrial septum being closed, indicating that in those patients the PFO was not the mechanism of the thromboembolic event in the first place. PMID- 15804808 TI - Abstracts from VII Swedish Heart Association Meeting and the XX Nordic Congress of Cardiology. Malmo, Sweden, April 27-29, 2005. PMID- 15804807 TI - Platelet scintigraphy results of heparin versus streptokinase treatment in acute deep vein thrombosis. AB - OBJECTIVE: The aim of the study was to document the therapeutic effects and primary recanalization ratios of either streptokinase or heparin in the therapeutic process of deep vein thrombosis (DVT). The results are obtained with the help of platelet scintigraphy. DESIGN: The investigation was planned as a randomized, controlled and prospective study, including 20 patients within two groups. Group I contained 10 patients undergoing IV infusion therapy of unfractionated heparin while Group II contained 10 patients undergoing thrombolytic therapy using streptokinase. After 8 weeks, scintigraphies using Tc 99m HMPAO-labeled thrombocytes were made, and interpreted by two blinded nuclear medicine physicians. The results are compared using Mann-Whitney U- and chi2 tests. RESULTS: No complication leading to interruption of therapy took place. Significantly smaller pathologic thrombocyte accumulations were detected in the streptokinase group. The difference was statistically significant (p = 0.019). CONCLUSION: Although a general clinical improvement had been observed in all patients, the scintigraphic investigation showed that the thrombolytic therapy maintained a better deep vein luminous patency. PMID- 15804809 TI - Theoretical predictions of chemical degradation reaction mechanisms of RDX and other cyclic nitramines derived from their molecular structures. AB - Analysis of environmental degradation pathways of contaminants is aided by predictions of likely reaction mechanisms and intermediate products derived from computational models of molecular structure. Quantum mechanical methods and force field molecular mechanics were used to characterize cyclic nitramines. Likely degradation mechanisms for hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) include hydroxylation utilizing addition of hydroxide ions to initiate proton abstraction via 2nd order rate elimination (E2) or via nucleophilic substitution of nitro groups, reductive chemical and biochemical degradation, and free radical oxidation. Due to structural similarities, it is predicted that, under homologous circumstances, certain RDX environmental degradation pathways should also be effective for octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) and similar cyclic nitramines. Computational models provided a theoretical framework whereby likely transformation mechanisms and transformation products of cyclic nitramines were predicted and used to elucidate in situ degradation pathways. PMID- 15804810 TI - Investigation of anticancer activity of macrocyclic Schiff bases by means of 4D QSAR based on simplex representation of molecular structure. AB - Influence of the molecular structure of macrocyclic pyridinophanes, their analogues and some other compounds on anticancer activity (Leukemia, central nervous system (CNS) cancer, prostate cancer, breast cancer, melanoma, non-small cell lung cancer, colon cancer, ovarian cancer, renal cancer) was investigated by means of a new 4D-QSAR approach based on the simplex representation of molecular structures (SiRMS). For all the investigated molecules, the 3D structural models were first created and the set of conformers (fourth dimension) was used. Each conformer was represented as a system of different simplexes (tetratomic fragments of fixed structure, chirality and symmetry). Statistic characteristics of the QSAR partial least squares (PLS) models were satisfactory (correlation coefficient r=0.990-0.861; cross-validation coefficient CVR=0.914-0.633). The molecular fragments increasing and decreasing anticancer activity were defined. This information may be useful for the design and direct synthesis of novel anticancer agents. PMID- 15804811 TI - Highly diverse, massive organic data as explored by a composite QSPR strategy: an advanced study of boiling point. AB - An improved strategy of quantitative structure-property relationship (QSPR) studies of diverse and inhomogeneous organic datasets has been proposed. A molecular connectivity term was successively corrected for different structural features encoded in fragmental descriptors. The so-called solvation index 1chis (a weighted Randic index) was used as a "leading" variable and standardized molecular fragments were employed as "corrective" class-specific variables. Performance of the new approach was illustrated by modelling a dataset of experimental normal boiling points of 833 organic compounds belonging to 20 structural classes. Firstly, separate QSPR models were derived for each class and for eight groups of structurally similar classes. Finally, a general model formed by combining all the classes together was derived (r2=0.957, s=12.9degreesC). The strategy outlined can find application in QSPR analyses of massive, highly diverse databases of organic compounds. PMID- 15804812 TI - Assessment of baseline toxicity of mono-cyclic aromatic compounds by pseudomonas initial oxygen uptake assay. AB - The objective of this study was to develop quantitative structure-activity relationships (QSARs) for the toxicity of mono-cyclic aromatic compounds in the Pseudomonas putida initial oxygen uptake assay. The QSARs were developed using response-surface based on descriptors for chemical hydrophobicity (logP) and electrophilicity (LUMO). The model log (Ki(-1)=0.434 (+/-0.011) log P-0.389 (+/ 0.013) LUMO - 2.13(+/-0.031); n=155, r2=0.941, r2(adj)=0.940, s=0.119, F=1206 led us to conclude that the polar and non-polar narcotics were statistically indistinguishable. Pentafluorophenol, pentachlorophenol and most dinitrophenols classified as weak acid respiratory uncouplers in literature fit well into this model when they were treated as their corresponding phenoxides. This latter result suggests that the action mechanism of these phenols should be reevaluated. PMID- 15804813 TI - Predictability and prediction of lowest observed adverse effect levels in a structurally heterogeneous set of chemicals. AB - A database of chronic lowest observed adverse effect levels (LOAELs) for 234 compounds, previously compiled from different sources (Toxicology Letters79, 131 143 (1995)), was modelled using graph theoretical descriptors. This study reveals that data are not homogeneous. Only those data originating from the U.S. Environmental Protection Agency (EPA) reports could be well modelled by multilinear regression (MLR) and linear discriminant analysis (LDA). In contrast, data available from the specific procedures of the National Toxicology Program (NTP) database introduced noise and did not render good models either alone, or in combination with the EPA data. PMID- 15804814 TI - Development of an information-intensive structure-activity relationship model and its application to human respiratory chemical sensitizers. AB - Structure-activity relationship (SAR) models are recognized as powerful tools to predict the toxicologic potential of new or untested chemicals and also provide insight into possible mechanisms of toxicity. Models have been based on physicochemical attributes and structural features of chemicals. We describe herein the development of a new SAR modeling algorithm called cat-SAR that is capable of analyzing and predicting chemical activity from divergent biological response data. The cat-SAR program develops chemical fragment-based SAR models from categorical biological response data (e.g. toxicologically active and inactive compounds). The database selected for model development was a published set of chemicals documented to cause respiratory hypersensitivity in humans. Two models were generated that differed only in that one model included explicate hydrogen containing fragments. The predictive abilities of the models were tested using leave-one-out cross-validation tests. One model had a sensitivity of 0.94 and specificity of 0.87 yielding an overall correct prediction of 91%. The second model had a sensitivity of 0.89, specificity of 0.95 and overall correct prediction of 92%. The demonstrated predictive capabilities of the cat-SAR approach, together with its modeling flexibility and design transparency, suggest the potential for its widespread applicability to toxicity prediction and for deriving mechanistic insight into toxicologic effects. PMID- 15804815 TI - Quantifying hydrogen bonding in QSAR and molecular modeling. AB - Quantitative descriptions of hydrogen bonding for use in QSAR and molecular modeling by means of H-bond descriptors have been analyzed in detail in this paper. Ten new H-bond surface and enthalpy integral descriptors were proposed. The usefulness of these new descriptors, as well as previously developed descriptors was verified using a set of 154 drugs for which data for intestinal absorption in humans were available. The results showed that descriptors such as the number of H-bond acceptor and donor atoms and polar surface area (PSA) did not sufficiently describe the actual H-bonding ability of atoms in molecules. Thus, to enable successful modeling it was necessary to introduce descriptors directly related to the experimental thermodynamics of hydrogen bonding. PMID- 15804820 TI - Revised BYLAWS of the Biological Stain Commission. PMID- 15804821 TI - Determination of optimal rehydration, fixation and staining methods for histological and immunohistochemical analysis of mummified soft tissues. AB - During an excavation headed by the German Institute for Archaeology, Cairo, at the tombs of the nobles in Thebes-West, Upper Egypt, three types of tissues from different mummies were sampled to compare 13 well known rehydration methods for mummified tissue with three newly developed methods. Furthermore, three fixatives were tested with each of the rehydration fluids. Meniscus (fibrocartilage), skin, and a placenta were used for this study. The rehydration and fixation procedures were uniform for all methods. The stains used were standard hematoxylin and eosin, elastica van Gieson, periodic acid-Schiff, and Grocott, and five commercially obtained immunohistochemical stains including pancytokeratin, vimentin, alpha-smooth-muscle-actin, basement membrane collagen type IV, and S 100 protein. The sections were examined by transmitted light microscopy. Our study showed that preservation of the tissue is dependent on the quality and effectiveness of the combination of the rehydration and fixation solutions, and that the quality of the histological and histochemical stains is dependent on the tissue quality. In addition, preservation of the antigens in the tissues is dependent on tissue quality, and fungal permeation had no influence on the tissue. Finally, the results are tissue specific. For placenta the best solution combination was Sandison and solution III (both fixed with formaldehyde) while results for skin were best with Ruffer I (using formaldehyde and Schaffer as fixatives), Grupe et al. (using formaldehyde as a fixative) and solution III (in combination with formaldehyde and Bouin fixatives). Ruffer II (using formaldehyde as a fixative) and solution III (in combination with Schaffer fixative) gave the best results for fibrocartilage. PMID- 15804822 TI - Staining human lymphocytes and onion root cell nuclei with madder root. AB - We performed staining experiments on cells using natural dyes and different mordants using techniques that are used for wool and silk dyeing. The natural dye sources were madder root, daisy, corn cockle and yellow weed. Ferrous sulfate, copper sulfate, potassium tartrate, urea, potassium aluminum sulfate and potassium dichromate were used as mordants. Distilled water, distilled water plus ethanol, heptane, and distilled water plus methanol were used as solvents. All dye-mordant-solvent combinations were studied at pH 2.4, 3.2 and 4.2. The generic staining procedure was to boil 5-10 onion roots or stimulated human lymphocyte (SHL) preparations in a dye bath on a hot plate. Cells were examined at every half hour. For multicolor staining, madder-dyed lymphocytes were decolorized, then stained with Giemsa. The AgNOR technique was performed following the decolorization of Giemsa stained lymphocytes. Good results were obtained for both onion root cells and lymphocytes that were boiled for 3 h in a dye bath that included 4 g madder root, 4 g ferrous sulfate as mordant in 50 ml of 1:1 (v/v) methanol:distilled water. The pH was adjusted to 4.2 with 6 ml acetic acid. We conclude that madder root has potential as an alternative dye for staining biological materials. PMID- 15804823 TI - Immunoreactivity of gastric ECL and A-like cells in fasted and fed rats and mice. AB - The oxyntic mucosa of rat and mouse stomach harbors histamine-producing ECL cells and ghrelin-producing A-like cells. The ECL cells are known to be active when the circulating gastrin levels are elevated in response to food intake. The A-like cells are the main source of circulating ghrelin. In response to starvation, the circulating ghrelin is elevated as a hunger signal. The aim of the present work was to study the correlation between the immunoreactivities and cellular activities of the ECL cells and A-like cells. Rats were either fed or fasted for 48 h and mice for 24 h. Immunohistochemical examination with antiserum against chromogranin A-derived fragment pancreastatin revealed both the ECL cells and the A-like cells without a difference between fasted and fed animals. Histamine was limited to the ECL cells with no significant difference between fasted and fed animals. Histidine decarboxylase (HDC) immunoreactivity occurred predominately in the ECL cells of the fed, but not fasted, animals in which the HDC enzymatic activity in the oxyntic mucosa was higher than in fasted animals. Ghrelin immunoreactivity was increased in terms of intensity, but not cell density in fasted animals. Thus, the immunoreactivities of ECL cells and A-like cells might be affected by starvation. PMID- 15804824 TI - Distribution of rDNA in the nucleus of Giardia lamblia: detection by Ag-I silver stain. AB - Previous investigations have proved that diplomonads have primitive cell nuclei and lack a nucleolus. We determined the distribution of ribosomal DNA (rDNA) in diplomonad nuclei that lacked a nucleolus. Giardia lamblia was used as the experimental organism with Euglena gracilis as the control. The distribution of rDNA was demonstrated indirectly by the modified Ag-I silver technique that can indicate specifically the nucleolus organizing region (NOR) by both light and electron microscopy. In ultrathin sections of silver stained Euglena cells, all silver grains were concentrated in the fibrosa of the nucleolus, while no silver grains were found in the cytoplasm, nucleoplasm, condensed chromosomes or pars granulosa of the nucleus. In the silver stained Giardia cells, no nucleolus was found, but a few silver grains were scattered in the nucleus. This suggests that the rDNA of Giardia does not form an NOR-like structure and that its nucleus is in a primitive state. PMID- 15804826 TI - Alkaline phosphatase method for use with resinous mounting media. PMID- 15804825 TI - Improvements in histological quality and signal retention following in situ hybridization in early chick embryos using plastic resin and recolorization. AB - We describe a novel method that allows reliable detection of in situ hybridization signals in thin sections of plastic embedded embryos. Sections from plastic embedded embryos are thinner and have superior histological quality compared to paraffin, gelatin, agarose embedded sections or cryosections; however, plastic resin traditionally has not been used as an embedding medium following in situ hybridization because of loss of signal. When signal is detected with alkaline phosphatase and NBT/BCIP, the resulting colored precipitate is subject to fading when samples are exposed to organic compounds. The colored precipitate can be redeposited by repeating the NBT/BCIP reaction following plastic sectioning. This recolorization shows no loss of specificity, because signal is detected only where the anti-digoxigenin/alkaline phosphatase conjugated antibody is bound to the riboprobe. Strong signals can be detected without recolorization; however, weaker signals require the recolorization step. This novel method of re-depositing colored precipitate after processing and sectioning allows accurate determination of the location of gene expression and study of this expression in high quality histological sections of early chick embryos. PMID- 15804828 TI - Nerve globins in invertebrates. AB - The expression of nerve hemoglobins in invertebrates is a well-established fact, but this occurrence is uncommon. In the species where nerve globins occur, they probably function as an oxygen store for sustaining activity of the nerves during anoxic conditions. Although invertebrate nerve globins are functionally similar with respect to O2 affinity, they are by no means uniform in structure and can differ in size, cellular localization and heme-coordination. The best-studied nerve globin is the mini-globin of Cerebratulus lacteus, which belongs to a class of globins containing the polar TyrB10/GlnE7 pair in the distal pocket. The amide and phenol side chains normally cause low rates of O2 dissociation and ultra-high O2 affinity by forming strong hydrogen bonds with bound ligands. Cerebratulus hemoglobin, however, has a moderate O2 affinity, due to the presence of a third polar amino-acid in its active site, ThrE11, which inhibits hydrogen bonding to bound oxygen by the B10 tyrosine side chain. PMID- 15804829 TI - Reversible hexa- to penta-coordination of the heme Fe atom modulates ligand binding properties of neuroglobin and cytoglobin. AB - Neuroglobin (Ngb) and cytoglobin (Cygb) are two recently discovered intracellular members of the vertebrate hemoglobin (Hb) family. Ngb, predominantly expressed in nerve cells, is of ancient evolutionary origin and is homologous to nerve-globins of invertebrates. Cygb, present in many different tissues, shares common ancestry with myoglobin (Mb) and can be traced to early vertebrate evolution. Ngb is held to facilitate O2 diffusion to the mitochondria and to protect neuronal cells from hypoxic-ischemic insults, may be an oxidative stress-responsive sensor protein for signal transduction, and may carry out enzymatic activities, such as NO/O2 scavenging. Cygb is linked to collagen synthesis, may provide O2 for enzymatic reactions, and may be involved in a ROS(NO)-signaling pathway(s). Ngb and Cgb display the classical three-over-three alpha-helical fold of Hb and Mb, and are endowed with a hexa-coordinate heme-Fe atom, in their ferrous and ferric forms, having the heme distal HisE7 residue as the endogenous ligand. Reversible hexa- to penta-coordination of the heme Fe atom modulates ligand binding properties of Ngb and Cygb. Moreover, Ngb and Cygb display a tunnel/cavity system within the protein matrix held to facilitate ligand channeling to/from the heme, multiple ligand copies storage, multi-ligand reactions, and conformational transitions supporting ligand binding. PMID- 15804830 TI - Tracing the structure-function relationship of neuroglobin and cytoglobin using resonance Raman and electron paramagnetic resonance spectroscopy. AB - The physiological role of neuroglobin and cytoglobin, two vertebrate globins discovered in the last 5 years, is not yet clearly understood. In this work, we review the structural information on these globins and its implication on the possible protein function, obtained by electron paramagnetic resonance and resonance Raman spectroscopy. All studies reveal a high flexibility in the heme pocket region of neuroglobin. Together with the observation that the distal ligand of the heme iron is the endogenous E7-histidine in both the ferric and ferrous form of neuroglobin and cytoglobin, the flexibility of the heme environment in neuroglobin will play a crucial role in the globins' ability to bind and stabilize exogenous ligands. PMID- 15804831 TI - The cellular and subcellular localization of neuroglobin and cytoglobin -- a clue to their function? AB - Neuroglobin and cytoglobin are recently discovered respiratory proteins of vertebrates with yet ill-defined physiological functions. Neuroglobin is widely expressed in neurons, but not glia, in the vertebrate central and peripheral nervous systems. Other major expression sites are the retina and endocrine tissues. This distribution is indicative of a function of neuroglobin in metabolically most active, oxygen-consuming cell types, but does not yet allow to safely distinguish between different cellular roles, such as oxygen homeostasis, scavenging of reactive oxygen species or sustaining energy metabolism. Cytoglobin is predominantly expressed in connective tissue fibroblasts and related cell types in the body organs. Its main function may therefore be related to the specific amounts of extracellular matrix. Cytoglobin may hypothetically be involved in the oxygen-consuming maturation of collagen proteins. Cytoglobin is also expressed in distinct cell types of brain and retina. Its distribution strikingly differs from neuroglobin, suggesting an independent, yet unknown function. PMID- 15804832 TI - Hypoxia/ischemia and the regulation of neuroglobin and cytoglobin expression. AB - In analogy to hemoglobin (Hb) and myoglobin (Mb), neuroglobin (Ngb) and cytoglobin (Cygb) are supposed to be involved in oxygen (O2) storage and delivery. The Cygb gene harbours both conserved HREs and mRNA stabilization sites, strongly suggestive of an oxygen-dependent regulation. We examined the relative transcriptional changes of Ngb and Cygb in a situation of chronic hypoxia using real-time quantitative PCR. We could conclude that Cygb is a hypoxia-induced gene, which is transcriptionally upregulated during chronic hypoxia in a hippocampal neuronal cell line and in multiple murine metabolically active tissues. The mechanism of induction of Cygb is HIF-1alpha dependent. HIF-1 is unique among mammalian transcription factors with respect to the specificity and sensitivity of its induction by hypoxia. Ngb expression seems to be regulated using other response elements and is less influenced by hypoxia. PMID- 15804833 TI - Functional properties of neuroglobin and cytoglobin. Insights into the ancestral physiological roles of globins. AB - Neuroglobin and cytoglobin are two recently discovered vertebrate globins, which are expressed at low levels in neuronal tissues and in all tissues investigated so far, respectively. Based on their amino acid sequences, these globins appear to be phylogenetically ancient and to have mutated less during evolution in comparison to the other vertebrate globins, myoglobin and hemoglobin. As with some plant and bacterial globins, neuroglobin and cytoglobin hemes are hexacoordinate in the absence of external ligands, in that the heme iron atom coordinates both a proximal and a distal His residue. While the physiological role of hexacoordinate globins is still largely unclear, neuroglobin appears to participate in the cellular defence against hypoxia. We present the current knowledge on the functional properties of neuroglobin and cytoglobin, and describe a mathematical model to evaluate the role of mammalian retinal neuroglobin in supplying O2 supply to the mitochondria. As shown, the model argues against a significant such role for neuroglobin, that more likely plays a role to scavenge reactive oxygen and nitrogen species that are generated following brain hypoxia. The O2 binding properties of cytoglobin, which is upregulated upon hypoxia, are consistent with a role for this protein in O2 requiring reactions, such as those catalysed by hydroxylases. PMID- 15804834 TI - Genome-wide analysis of the globin gene family of C. elegans. AB - The aim of our study was to annotate sequences for 35 putative globins from the nematode Caenorhabditis elegans. All these proteins are expressed, but seven of these differ from the gene predictions in Wormbase. The entire polypeptide sequences for 31 genes and the core globin domain of four proteins were confirmed or corrected. All core globin domains were aligned manually following a procedure that was designed to fit the putative sequences to the crystal structure based alignment of 56 known globin crystal structures. Neighbor-joining analysis of the resulting alignment showed that the majority of these globins are very divergent from each other, possibly suggesting a long evolutionary divergence. The surprisingly high number and low sequence conservation of putative globins in this small organism urges a detailed functional analysis. PMID- 15804835 TI - Neuroglobin and cytoglobin: genes, proteins and evolution. AB - Hemoglobin and myoglobin are oxygen transport and storage proteins of most vertebrates. Neuroglobin (Ngb) and cytoglobin (Cygb)--two recent additions to the vertebrate globin superfamily--have still disputed functions. Combining the data from all available resources, we investigate the evolution of these novel globins. Both Ngb and Cygb show little sequence variation in vertebrate evolution, suggesting conserved structures and functions, and an important role in the animal's metabolism. Exon-intron patterns remained unchanged in Ngb and Cygb, with the exception of the addition of a 3' exon to Cygb early in mammalian evolution. In phylogenetic analyses, Ngb forms a common branch with globin X, another recently identified globin with undefined function in lower vertebrates, and with some invertebrate nerve globins. This shows an early divergence of this branch in animal evolution. Cygb is related to myoglobin, and associated with an eye-specific globin from birds. The pattern of globin evolution shows that proteins with clear respiratory roles evolved independently from intracellular globins with uncertain functions. This result suggests either multiple independent functional changes or a yet undefined respiratory role of tissue globins like Ngb and Cygb. PMID- 15804836 TI - Neuroglobin ligand binding kinetics. AB - Neuroglobin, cytoglobin, and hemoglobins from Drosophila melanogaster and Arabidopsis thaliana were studied for their ligand binding properties versus temperature. These globins have a common feature of being hexacoordinated (via the distal histidine) under deoxy conditions, displaying and enhanced amplitude for the alpha absorption band at 560 nm. External ligands can bind, but the transition from the hexacoordinated form to the ligand (L) bound species is slow, as expected for a replacement reaction Fe-His <--> Fe <--> Fe-L. Histidine binding is on the order of 1 ms; dissociation times are variable, and may be as long as 1 s for the highest histidine affinities. Oxygen binds rapidly but dissociates slowly, requiring as much as 1 s. These rates would correspond to a very high affinity for the pentacoordinated form; however, competition with the distal histidine leads decreases the affinity for the external ligand. The observed oxygen affinity remains in the range of 1 to 10 mm Hg. The low oxygen dissociation indicates a stabilization via H-bonds as for certain globins from parasites (Ascaris, the trematodes). Other ligands such as CO, or CN for the ferric form, show a decreased affinity, since only the competition with the E7 histidine, but not the stabilizing H-bond, plays a role. In addition, the competitive internal ligand leads to a weaker observed temperature dependence of the ligand affinity, since the difference in equilibrium energy for the two ligands is much lower than that of ligand binding to pentacoordinated hemoglobin. This effect could be of biological relevance for certain organisms, since it would lead to an oxygen affinity that is nearly independent of temperature. PMID- 15804838 TI - Stem cell therapy for retinal degeneration: retinal neurons from heterologous sources. AB - Over the past few years a great deal of interest has been generated in using stem cells/progenitors to treat degenerative diseases that afflict different tissues, including retina. This interest is due to the defining properties of stem cells/progenitors, the ability of these cells to self-renew and generate all the basic cell types of the particular tissue to which they belong. In addition, the recent reports of plasticity of the adult tissue-specific stem cells/progenitors and directed differentiation of the embryonic cells (ES) has fueled the hope for cell and gene therapy using stem cells from heterologous sources. Will this approach work for treating retinal degeneration? Here, we review the current state of knowledge about obtaining retinal cells from heterologous sources, including ES cells. PMID- 15804839 TI - Photoreceptors derived from adult iris tissue: prospects for retinal transplantation. AB - While a number of retinal transplantation studies using various types of donor cells have been performed thus far, our study focused on iris tissue as a donor cell source. This is because donor cells from iris pigment epithelium have the following characteristics: (1) they are embryonically related to the neural retina; (2) autologous iris tissue can be obtained via a surgical approach; and (3) they can be cultured to increase the number of donor cells and establish photoreceptor-like cells from iris-derived cells by means of the appropriate gene transfer. Although the potential of iris-derived cells has been indicated, there remain many issues to be investigated. PMID- 15804840 TI - Embryonic stem cells: potential source for ocular repair. AB - Many ocular diseases, such as retinitis pigmentosa and age-related macular degeneration, reflect damage to specific cells that are not normally repaired or replaced. Preliminary results of animal studies suggest that these degenerative diseases may be treatable by transplantation of healthy fetal cells. However, obtaining a sufficient number of suitable donor cells remains a problem. The isolation of human embryonic stem (ES) cells has drawn much attention because of their potential clinical application for patients with these degenerative diseases. Because ES cells have the potential to generate all adult cell types, ocular diseases resulting from the failure of specific cell types would be potentially treatable through the transplantation of differentiated cells derived from ES cells. In addition, because ES cells can proliferate indefinitely in their undifferentiated state, they are expected to alleviate the problem of the shortage of donor cells for cell-replacement therapy. Recently, reproducible and efficient differentiation methods for the generation of lens cells, retinal neurons, and retinal pigment epithelial (RPE) cells from ES cells have been developed. This review focuses especially on these ocular cells differentiated from ES cells. We will also discuss the potential therapeutic uses of ES cells for the treatment of ocular diseases. PMID- 15804841 TI - Transcriptional factors involved in photoreceptor differentiation. AB - Regenerative medicine constitutes a potentially promising therapy for blind people suffering from retinal degenerative diseases such as retinitis pigmentosa and age-related macular degeneration. For the realization of retinal regeneration, it is necessary to establish 1) a method to produce functional photoreceptor cells in vitro and 2) successful transplantation of the donor cells to connect their axons to the recipient secondary neurons so that they can function properly. The results of experimental transplantation of human retinal photoreceptor cells from cadaveric eyes or of fetal retinal cells into the retina of RP patients have not been satisfactory, but encouraging enough to indicate that the transplantation of developing retinal cells may have beneficial results. Recently, attempts have been made to generate photoreceptor-like cells from stem cells, but it remains to be seen whether they are in fact photoreceptor cells. It is therefore important to fully understand the mechanisms involved in the development of these cells, and to characterize them not only by transcriptome but also by functional analysis. PMID- 15804842 TI - Transplantation of neuroblastic progenitor cells as a sheet preserves and restores retinal function. AB - Diseases affecting the outer retina are incurable once photoreceptors are lost, and these diseases usually cause retinal pigment epithelium (RPE) dysfunction. However, the inner retina can remain functional for some time, even though retinal remodeling occurs as compensation for photoreceptor loss. If the damaged part can be replaced with neuroblastic progenitor and RPE cells as sheets with a beneficial effect on function, vision loss may be prevented and vision may be restored. This review presents an overview of the research of transplanting sheets of neural retina, with or without its RPE, to the subretinal space. In different animal models of retinal degeneration, retinal transplants can morphologically reconstruct a damaged retina, and restore visual sensitivity. Good morphological integration of transplants with the host retina can occur, whereas other transplants exhibit a glial barrier. Synaptic connections between transplant and host have been indicated by transsynaptic tracing. Retinal transplants can restore and preserve visual responses in a small area of the superior colliculus corresponding to the placement of the transplant in the retina. The beneficial effect of retinal transplantation likely involves two mechanisms: trophic effects, e.g., rescue of host cones; and synaptic connectivity between transplant and host retina. PMID- 15804843 TI - Retinal vascular regeneration. AB - We discuss the potential use of stem cells for therapeutic angiogenesis in the treatment of retinal diseases. We demonstrate that the clinical utility of these EPC may be not limited in the treatment of ischemic retinal diseases but may also have application for the treatment of retinal degenerative disorders and for a form of cell-based gene therapy. One of the greatest potential benefits of bone marrow derived EPC therapy is the possible use of autologous grafts. Nonetheless, potential toxicities and unregulated cell growth will need to be carefully evaluated before this approach is brought to the clinics. PMID- 15804844 TI - Anthropometric criteria for the design of tractor cabs and protection frames. AB - Improved human-tractor interface designs, such as well-accommodated operator enclosures (i.e. cabs and protection frames) can enhance operator productivity, comfort and safety. This study investigated farm-worker anthropometry and determined the critical anthropometric measures and 3-D feature envelopes of body landmarks for the design of tractor operator enclosures. One hundred agriculture workers participated in the study. Their body size and shape information was registered, using a 3-D full-body laser scanner. Knee height (sitting) and another eight parameters were found to affect the cab-enclosure accommodation rating and multiple anthropometric dimensions interactively affected the steering wheel and gear-handle impediment. A principal component analysis has identified 15 representative human body models for digitally assessing tractor-cab accommodation. A set of centroid coordinates of 34 body landmarks and the 95% confidence semi-axis-length for each landmark location were developed to guide tractor designers in their placement of tractor control components in order to best accommodate the user population. Finally, the vertical clearance (90 cm) for agriculture tractor enclosure in the current SAE International J2194 standard appeared to be too short as compared to the 99th percentile sitting height of male farm workers in this study (100.6 cm) and in the 1994 National Health and Nutrition Examination Survey III database (99.9 cm) and of the male civilian population in the 2002 Civilian American and European Surface Anthropometric Resource database (100.4 cm). PMID- 15804845 TI - Composite scales of morningness and preferences: preliminary validity data in Peruvian undergraduates. AB - The aim of this study is to offer preliminary results about the validity of the composite morningness scale (CS) and the early/late preferences scale (PS) in a Peruvian sample. The relationship of both scales with the preferred rising and retiring times was analysed, along with the level of self-reported alertness. In Bohle et al.'s (2001) work, the relationship between morningness and preferred rising and retiring times was higher over the weekend than on weekdays. This difference explained the dispositional nature of morningness, due to the possible lesser influence of time schedules over the weekend in individuals' preferences. This result is replicated in a group of 139 Peruvian undergraduates, aged between 18 and 29 years (M = 21.73), of whom 78.4% were women. The relationship between morningness and (actual) normal rising and retiring times on weekdays and over the weekend is considered. The results partially confirm Bohle et al.'s (2001) hypothesis about preferred rising and retiring times and their relationship with the PS, and actual rising time and its relationship with the CS and PS. The differences in the level of self-reported alertness between morning, intermediate and evening-oriented groups provide support for the validity of both scales. Finally, the scores of CS and PS in Peruvian undergraduates are similar to those found by Smith et al. (2002) in university students from six countries. PMID- 15804846 TI - The effect of asymmetry on psychophysical lifting capacity for three lifting types. AB - The effect of asymmetry on a person's lifting capacity was investigated using the psychophysical approach. Ten male college students lifted a box from pallet height (15 cm) to conveyor height (75 cm) at a frequency of one and five lifts/min. Three types of asymmetric lifting tasks (step-turn, middle twist and twist) were studied using 90 and 180 degrees task angles. Lifting capacity reductions for middle twist and twist at a 90 degrees asymmetric angle were about one-half of the 30% reduction that would be calculated by the 1991 National Institute for Occupational Safety & Health (NIOSH) lifting equation. The lifting capacity reduction for step-turn at 180 degrees was 14.9%, although that reduction cannot be calculated in the NIOSH equation. The middle twist lifting capacity was greatest among the three types at a 90 degrees task angle. The reductions for the middle twist and step-turn were not proportional to the task angle. This is contrary to the proportional reduction in the NIOSH lifting equation. Heart rate did not increase with an increase in task angle. Based on the results of this research, a different approach to assigning the asymmetric multiplier is proposed. This approach includes a task angle (as opposed to asymmetric angle) of up to 180 degrees. PMID- 15804847 TI - Modulation of mechanical and muscular load by footwear during catering. AB - The BGN (Berufsgenossenschaft Nahrungsmithl und Gaststatten) reports 70% of job induced days off work to be connected with traumas of the ankle joint or overloading of the leg, knee and lower back, with an increased incidence in service areas outdoors (R. Grieshaber, personal communication). Workspace environments usually contain narrow passages, slopes or stairs and sudden changes between different surfaces. The aim of this study was to investigate the biomechanical load on the lower extremity and the low back during catering service when wearing different types of footwear. Thus, the potential for altering mechanical stress experienced during catering by variations in footwear was explored. Sixteen experienced waiters followed a course typical for a combined indoor-outdoor service area. Three different types of footwear were investigated using pressure distribution measurements, rearfoot goniometry and electromyography. A discriminant analysis revealed that the factors subject, shoe and surface affect rear foot movement or pressure distribution in different ways. A MANOVA demonstrated significant differences in loading parameters between footwear types. In general, these differences increased in magnitude in critical situations, such as climbing stairs or crossing slippery surfaces. The results of this study demonstrate that manipulations to footwear offer a great potential for modulating loads experienced during catering. Based on the results, the effects of constructional features are discussed. The method proposed can be applied to evaluate shoe modifications under realistic workplace conditions. PMID- 15804848 TI - Effect of button location on driver's visual behaviour and safety perception. AB - Although buttons are a frequently used type of control in car interiors, little research has been undertaken on how the usage of buttons affects the visual behaviour of the driver. In this study, the aim was to analyse the effect of push button location and auditory feedback on drivers' visual time off road and safety perception when driving in a real traffic situation. The effect of six button locations (five on the centre stack, one near the gear stick) was tested. Drivers' visual behaviour was studied in real traffic on a motorway. An eye tracking system recorded the visual behaviour of eight drivers who, in 96 repeated trials each, were instructed to press a specific button. Data analysis focused on the drivers' visual time off road and safety perception in relation to the location of the button to be pressed. Auditory feedback did not show a significant effect on visual time off road. The time off road increased significantly as the angle increased between the normal line of sight and button location for the five buttons placed on the centre stack. Results for the button located close to the gear stick, with the highest eccentricity, produced a short time off road. This unexpected finding is discussed in terms of three potential explanations: 1) the role of perceptual discrimination; 2) risk perception; and 3) motor control. PMID- 15804849 TI - Effects of antagonistic co-contraction on differences between electromyography based and optimization based estimates of spinal forces. AB - Estimates of spinal forces are quite sensitive to model assumptions, especially regarding antagonistic co-contraction. Optimization based models predict co contraction to be absent, while electromyography (EMG) based models take co contraction into account, but usually assume equal activation of deep and superficial parts of a muscle. The aim of the present study was to compare EMG based and optimization based estimates of spinal forces in a wide range of work tasks. Data obtained from ten subjects performing a total of 28 tasks were analysed with an EMG driven model and three optimization models, which were specifically designed to test the effects of the above assumptions. Estimates of peak spinal forces obtained using the different modelling approaches were similar for total muscle force and its compression component (on average EMG based predictions were 5% higher) and were closely related (R > 0.92), while differences in predictions of the peak shear component of muscle force were more substantial (with up to 39% lower estimates in optimization based models, R > 0.79). The results show that neither neglecting antagonistic co-contraction, nor assuming equal activation of deep and superficial muscles, has a major effect on estimates of spinal forces. The disparity between shear force predictions was due to an overestimation of activity of the lateral part of the internal oblique muscle by the optimization models, which is explained by the cost function preferentially recruiting larger muscles. This suggests that a penalty for active muscle mass should be included in the cost function used for predicting trunk muscle recruitment. PMID- 15804853 TI - Automatic segmentation of medical images using image registration: diagnostic and simulation applications. AB - Automatic identification of the boundaries of significant structure (segmentation) within a medical image is an are of ongoing research. Various approaches have been proposed but only two methods have achieved widespread use: manual delineation of boundaries and segmentation using intensity values. In this paper we describe an approach based on image registration. A reference image is prepared and segmented, by hand or otherwise. A patient image is registered to the reference image and the mapping then applied to ther reference segmentation to map it back to the patient image. In general a high-resolution nonlinear mapping is required to achieve accurate segmentation. This paper describes an algorithm that can efficiently generate such mappings, and outlines the uses of this tool in two relevant applications. An important feature of the approach described in this paper is that the algorithm is independent of the segmentation problem being addresses. All knowledge about the problem at hand is contained in files of reference data. A secondary benefit is that the continuous three dimensional mapping generated is well suited to the generation of patient specific numerical models (e.g. finite element meshes) from the library models. Smoothness constraints in the morphing algorithm tend to maintain the geometric quality of the reference mesh. PMID- 15804854 TI - A fuzzy logic based closed-loop control system for blood glucose level regulation in diabetics. AB - In this study, a closed-loop system to control the plasma glucose level in patients with diabetes mellitus type 1 is proposed. This control scheme is based on fuzzy logic control theory to maintain a normoglycaemic average of 4.5 mmol 1( 1) and the normal conditions for free plasma insulin concentration in severe initial state; in particular, when the diabetic patient is subjected to a glucose meal disturbance or fluctuations in the measured glucose level due to error in the measuring instrument. The proposed controller has demonstrated superiority over other conventional controlling therapies. While fuzzy logic controllers have shown promising results in many fields, a comparative study is presented with well-known conventional controllers such as Proportional-Integral-Derivative (PID) and continuous insulin infusion control strategies. The simulated results, for the proposed controller, are presented and discussed. PMID- 15804855 TI - A kinetic model for simulation of blood coagulation and inhibition in the intrinsic path. AB - A mathematical simulation pathway for the generation of thrombin has been developed with various assumptions made of kinetic rate laws and their summation for reactions involving the activation of factors VIII, IX, X and V and protein C in the formation of thrombin. The object of the computational modelling study is to stimulate the activation and inhibition of blood coagulation. The level of complexity and assumed parameters makes conclusions uncertain. However, an interesting outcome is that kinetic rates may show oscillation behavior under particular high levels of protein C feedback inhibition. The model, which permits the assessment of the reaction over a broad range of conditions, would defy quantitative practical use, but could have predictive value as a qualitative descriptor of coagulation. PMID- 15804856 TI - A thermodynamic model to predict the thermal response of living beings during pneumoperitoneum procedures. AB - In this work, hypothermia associated with pneumoperitoneum procedures is studied. A thermodynamic model is developed to allow for the computational simulation of the thermal body response to pneumoperitoneum procedures, which are required by laparoscopic surgery. The numerical results predict the body temperature decay (or loss of energy) in time when the pneumoperitoneum procedures is conducted in patient. The influence of several operating parameters (e.g. inlet air mass flow rate and temperature) on the resulting hypothermia level is analysed. Therefore, the model allows the identification of parameters that have to be controlled to minimize the loss of energy, and consequently, the hypothermia level due to pneumoperitoneum procedures. PMID- 15804857 TI - Stimulated muscle force assessment of the sternocleidomastoid muscle in humans. AB - The aim of this study was to configure a force assessment device and determine potential testing protocols for quantitative evaluation of human neck muscles. The study design consisted of non-randomized control trials, with repeated measures; data from 12 normal subjects were obtained. Several apparatuses were designed, constructed and tested, i.e. single or short trains of supramaximal stimuli were used to activate sternocleidomastoid muscles in a seated position with strain gauges (6.2% variability with double-pulse stimulations) or in supine positions with load cells (5.2% variability with similar activation). Using a final configuration, maximum elicited peak forces were 1742 +/- 323 g for single pulse and 3976 +/- 484 g for double-pulse stimulations (n = 12). There were no significant differences in maximum recorded peak torques between sessions per individual. Yet, detectable muscle activities were simultaneously recorded in the contralateral sternocleidomastoid muscles. This non-invasive, quantitative assessment approach has novel value for determining treatment efficacy, disease progression, and/or approach has novel value for determining determining treatment efficacy, disease progression, and/or relative distribution of muscle strength in patients with abnormal neck muscle function. PMID- 15804858 TI - A quantitative gait assessment method based on energy exchange analysis during walking: a normal gait study. AB - In this paper a gait efficiency assessment method, Gait Energy Efficiency Index (GEEI), which can be used in evaluation of the progress of the rehabilitation process in disabled persons, is proposed and described. The method is based on calculation of cross correlation between normalized time courses of kinetic and potential energy of the body's centre of mass (COM). We hypothesized that GEEI in energetically optimal normal walking should be high and invariable of gait speed. The method was tested on twelve healthy subjects walking at three different speeds and contrasted to five established gait energy consumption assessment methods. The results showed that GEEI in normal walking is close to 1 regardless of walking speed. PMID- 15804859 TI - An improved windowing technique for heart rate variability power spectrum estimation. AB - Spectral analysis of heart rate variability (HRV) is an accepted method for assessment of cardiac autonomic function and its relationship to numerous disorders and diseases. Discrete Fourier transform (DFT) based methods are widely used for their easy applicability, computational speed and the possibility for direct interpretation of results. This study assesses the limitation of windowing of the RR interval series of power spectrum estimation using DFT for heart rate variability studies. The mean value of the RR interval series should be subtracted before windowing. This may leave a small residual DC component after windowing, but the RR interval series is properly tapered to zero at the beginning and end of the window. However, if the windowed RR interval series has a non-zero mean then subtracting this mean will create an abrupt transition between the first and last data points, and the padded zeros. This is equivalent to superimposing upon the RR interval series a rectangular pulse of the same length as the window, with a height equal to the subtracted mean value. In the present paper an approach to overcome the above effects of the window in reducing the signal energy and introducing the low frequency component into spectrum has been suggested and incorporated. Result have been compared for DC biasing of windowed data spectrum, bias of windowed data removed by substraction of mean data, and data processed to remove windowed mean level and to maintain mean power. Thus the preprocessing of RR interval series with this method improves the accuracy of HRV analysis methods. The study was carried out by smoothing the complete RR interval series by single Hann window and by 50% overlapping the data segments of 256 data points followed by the DFT. Overlapping the data segments provides equal weight to all values in the RR interval series and smoothed spectral estimate with clearly dominant peaks in low- and high-frequency regions. PMID- 15804863 TI - Decreased testicular expression of cAMP response element modulator (CREM) in rat with varicocele. AB - This study examines the hypothesis that varicocele would impair the testicular expression of cAMP response element modulator (CREM) in experimental rats. Thirty (30) rats were selected at random, of 20 were operated as varicocele's group; of 10 were for sham-operation as controlled group, testes were removed, fixed and stained in three months. Makler Score was adopted to analyze the bore, limitan's thickness, layer number of cell, the degree of cell's maturity and average score in 200 and 100 seminiferous tubulae, respectively. CREM, HSP60 was determinated by hybridization in situ, the difference between two groups were compared. In varicocele group, seminiferous tubule's bore was decreased (101 +/- 2.2) vs 146 +/- 4.1), limitan became thicker (3.5 +/- 0.1 vs 1.9 +/- 0.2), cell's layer number was reduced (3.0 +/- 0.2 vs 5.5 +/- 0.1), cell's maturity turned to disturbance (3.6 +/- 0.3 vs 4.9 +/- 0.1), the average score was lower than controlled group (8.5 +/- 0.6 vs 16.0 +/- 1.2), they had significant differences (P < 0.001). The testicular CREM expression was significantly lower in varicocele group than in controlled one (VG2.0 +/- 0.32, SoG3.90 +/- 0.32) (p < 0.001), which was located in spermatogenous cell to spermatocyte stage. However, HSP60 expression in VG was higher than in SoG (3.85 +/- 0.3 versus 2.1 +/- 0.32) with significant differences (P < 0.001), the expression located in spermatid. Varicocele could lead to lower testicular CREM expression and breeding sperm functional lesion. PMID- 15804862 TI - Control of spermatogenesis in primate and prospect of male contraception. AB - The present review is a summary of mechanisms of spermatogenesis in primates with emphasis on anti-spermatogenesis of testosterone (T), gossypol, and "testicular heat stress" for development of male contraception, Both FSH and testosterone stimulate all phases of spermatogenesis. FSH is capable of amplifying the population of the differential spermatogonia (B1, B2, B3 and B4) and controls the spermatogonia production rate, and, in synergy with testosterone, regulating spermatogenesis in adult monkeys. Pituitary FSH beta gene expression is governed by a feedback of Beta inhibin, which is a major component of the testicular negative feedback signals. Beta inhibin secreted by Sertoli cells is in turn inhibited by testosterone from Leydig cells under the control of LH. Disturbance of the normal interaction of pituitary FSH with Sertoli cell Beta inhibin is responsible for azoospermia or oligozoospermia induced by exogenous T. Three possible regimens of T, gossypol and "heat stress" have been suggested for male contraception. They act on different sites and stages of spermatogenesis in testis or sperm activity in epididymis. Apoptosis induced by testosterone occurs mainly at staged VII-VIII of spermatogenesis while that by testicular "heat stress" mostly occurs at stages I-IV and X-XII. Low dose of gossypol mainly influences the sperm activity in the epididymis although it also acts on testicular spermatids. PMID- 15804864 TI - Sex steroids and spermatogenesis in the African catfish (Clarias gariepinus). AB - Little is known about the physiological regulations of spermatogenesis of the African catfish (Clarias gariepinus), a species used as an experimental model for two decades. After a brief introduction to the cystic type of spermatogenesis in fish, the role of androgens and estrogens will be discussed, leading to a conclusion that androgens are required for spermatogenesis while their mode of action is poorly understood. In the cystic mode of spermatogenesis in fish, the Sertoli cells are formed cystic, in the somniferous lobules enclosing a single germ cell clone, providing suitable experimental models to address questions of relevance for vertebrate spermatogenesis in general. PMID- 15804865 TI - Does orchidopexy revert the histological alterations in epididymal and vas deferens caused by cryptorchidism? AB - Cryptorchidism is a pathological condition in which the testicles are retained in the abdominal cavity, resulting in atrophic seminiferous tubules. Some gross structural abnormalities and histological altercations have been described in the epididymis and vas deferens in humans with cryptorchidic testes. Orchidopexy surgery restores testicular spermatogenesis in experimental and clinical procedures, but it is still unclear whether histological changes in the epididymis and vas deferens caused by cryptorchidism may be reverted by orchidopexy. The aim of this study was to evaluated the histological changes in the epididymis and vas deferens following experimental uni- bilateral cryptorchidism in mature and immature mice, and to determine whether altercations could be reversed by orchidopexy. Young and adult C57 BL6 mice were randomized into three groups: control mice, bi/unilaterally cryptorchidic mice and bilaterally cryptorchidic mice with orchidopexy. After evaluation of testis, epididymis and vas deferens, there were no histological alterations in contralateral epididymis of mice unilaterally cryptorchidic. Ipsilateral epididymis of unilaterally cryptorchidic mice and epididymis from bilaterally cryptorchidic mice showed significant histological alterations. Orchuidopexy resorted normal spermatogenesis and the histological features of epididymis. It would appear that persistent male infertility clinically observed after orchidopexy could not be related to histological alteration in the testis and epididymis. Development and maintenance of the vas deferens seems to be controlled independently of the epididymis since it was not altered by cryptorchidism condition. PMID- 15804866 TI - Effect of an abnormal sperm chromatin structural assay (SCSA) on pregnancy outcome following (IVF) with ICSI in previous IVF failures. AB - A high DNA fragmentation index (DFI) when performing the sperm chromatin structural (SCSA) assay was claimed to be so specific for male subfertility that even IVF and ICSI did not result in live pregnancies. The present study was designed to corroborate or refute these findings. The SCSA test was performed on the male partner from couples failing to have a successful pregnancy despite at least 2 previous IVF attempts. In contrast to the aforementioned studies, ongoing pregnancies were found despite working with a group of recalcitrant patients. Nevertheless, a high DFI score was associated with a trend for lower ongoing pregnancy rates especially related to a high miscarriage rate. Other more recent studies seem to support our conclusions. A high DFI score should influence a patient to choose IVF as a therapeutic modality sooner, especially with ICSI. PMID- 15804867 TI - Chromatin remodeling in spermatids: a sensitive step for the genetic integrity of the male gamete. AB - Several causes of male infertility remain idiopathic. Recently, the condensed state of the sperm head has been demonstrated as a discriminating parameter for the assessment of male infertility. Altered DNA condensation is associated with an increase in DNA strand breakage so the genetic integrity of the male gamete is threatened. The origin of the DNA strand breaks in unknown. However, transient DNA strand breaks appear in the whole population of elongating spermatids during mid-spermiogenesis steps. Most likely, these transient breaks are required to support the change in DNA topology associated with chromatin remodeling at these steps. Histones hyperacetylation is also coincident with the DNA strand breakage steps. Hyperacetylation of histones may represent a necessary condition for strand breakages to form allowing access to the yet unknown enzymatic activity involved in the removal of DNA supercoils. A better characterization of this enzyme activity at these steps is necessary as this may represent a very sensitive process where altercations in the genetic integrity of the male gamete may arise and persist up to the mature spermatozoa. During the chromatin remodeling in spermatids, the combined DNA-condensing activities provides by the basic transition proteins and protamines may optimize the strand repair process emphasizing the link between altered sperm DNA condensation and DNA fragmentation. The mutagenic potential of these events may have been overlooked as it may result in fertility and/or developmental problems. PMID- 15804868 TI - Prostatic cancer/benign prostatic hypertrophy. Subcellular distribution of estradiol/androgen receptors. AB - Six microsomal population of estradiol and androgen receptors have been characterized in human benign prostatic hypertrophy (BPH) and prostate cancer (PCa). Estradiol receptor (ER) and androgen receptors (AR) were extracted using 0.6 M KCL and determined by the dextran-coated charcoal method. ER and AR levels were smaller in BPH plasma membranes (PM) than in Pca cases. For functions 3, 4, 6, the ER values in PCa were 25-38% less with regard to BPH ER values. Whereas in PCa, AR values obtained in all fractions were higher when compared to BPH AR values. In benign prostatic hypertrophy and prostatic cancer, ER and AR levels were significantly higher in the nuclear fraction. In the nuclear fraction, ER and AR levels in BPH and PCa were significantly different. The subcellular distribution of AR and ER in BPH and PCa constitutes a reservation mechanism and processing a receptors for their continued growth. PMID- 15804869 TI - Loupe-assisted modified one-layer vasovasostomy. AB - Several studies indicate that microsurgical modified one-layer vasovasostomy is comparable to the two-layer anastomosis with respect to patency and pregnancy rates. The objective of this study was to determine the feasibility and result of modified one-layer vasovasostomy under loupe magnification only. Thirty-two patients aged 28 to 64 years (mean 41.3 +/- 6 years) underwent vasovasostomy at CGMH from July 1997 to June 2002, with all operations being a modified on-layer anastomosis created with the aid of a 3 x loupe. The estimated duration of vasectomy ranged from 4 months to 27 years, with a mean of 9.2 +/- 4.8 years. Postoperative semen analysis and pregnancy were examined. Each patient was followed up at 1,4, and 12 weeks postoperatively. The total operation time ranged from 118 to 228 minutes (average 150 +/- 35 minutes). There was no operation related complication such as hematoma or wound infection. The patency rate was 89% (25/28), and the pregnancy rate at 2 years or more of follow-up was 39% (11/28). The patency and pregnancy rates were similar to those obtained in most studies of microsurgical vasovasostomy. For uncomplicated vasectomy reversal, this simple loupe-assisted modified one-lyer vasovasostomy seems to provide an adequate anastomosis. PMID- 15804870 TI - Relationship between mast cell and iNOS expression in testicular tissue associated with infertility. AB - The objective of this study was to investigate mast cells and iNOS expression in testis tissue, and to correlate these results with spermatogenic disorders. A total of 136 testicular biopsies were obtained from the testes of 80 patients with infertility. Their age ranged from 21 to 45 years. The biopsy specimens were immunohistochemically stained with antihuman tryptase for mast cells. In each section, all interstitial fields were evaluated for the total number of mast cells as well as the total number of Leydig cells. The number of mast cells per Leydig cell was calculated and recorded as mast cell index. Immunohistochemical iNOS staining was evaluated semiquantitatively according to intensity and the proportion of the stained cells. There was a significant increase of the mast cell index in all groups with testicular disorder compared with normal spermatogenesis group (p < 0.05). Increase of the index was in the order of hypospermatogenesis, maturation arrest and SCO, and index of SCO group was especially higher, i.e, more than twice than other groups. iNOS score was significantly higher in the SCO group than in the men with normal spermatogenesis, hypospermatogenesis, and maturation arrest (p < 0.05). Finally, a significantly statistical correlation was found between the iNOS score and mast cells index (r = 0.758, p = 0.001). Increase of mast cell index was observed in the groups of infertile testis, and high expression of iNOS in Leydig cells was associated with the highest mast cell index in SCO, the lesion with the most severe damage of the germ cell. PMID- 15804871 TI - Is varicocele a prognostic factor for determining sperm retrieval rate before testicular sperm extraction? AB - 66 nonobstructive azoospermic mean with normal genetic analysis composed of 32 (48%) patients with and 34 (52%) patients without varicocele were evaluated for the rate of sperm extraction five months after the varicocelectomy. Sperm retrieval was successful in 22 of 32 patients (68%) who had been operated because of varicocele and in 13 of 34 patients (38.2%) who had no varicocele (OR = 3.55) (CI: 1.15-11.27) (p = 0.025). Overall, sperm extraction was successful in 35 of 66 patients (53%). Repair of varicocele in non-obstructive azoospermic patients may return spermatogenesis to normal in spermatogenic focuses. So, the present of varicocele and its treatment might be considered as a prognostic factor for sperm retrieval in these patients. PMID- 15804872 TI - Are non-substance-related addictions really addictions? PMID- 15804873 TI - Desipramine treatment for cocaine dependence in buprenorphine- or methadone treated patients: baseline urine results as predictor of response. AB - The prognostic importance of baseline urines for cocaine was examined in a randomized, placebo-controlled, twelve-week clinical trial in 165 opioid- and cocaine-dependent patients who were treated with desipramine (DMI) in combination with buprenorphine (BUP) or methadone (Meth). Patients with a cocaine-positive urine at baseline (CU+) had significantly fewer cocaine-free urines than those with a negative urine at baseline (CU-neg). The CU+ patients showed a treatment effect of DMI. This DMI effect was significant in patients maintained on BUP but not on Meth. PMID- 15804874 TI - Mediating factors for the long-term effects of parental alcoholism in women: the contribution of other childhood stresses and resources. AB - The primary aim of this study was to identify the stresses and resources in childhood that mediate the relationship between parental alcoholism and adult outcomes in women. Adult outcomes included alcohol problems and measures of psychosocial adjustment. Standardized measures and a face-to-face interview were used to collect data on 290 community-dwelling women, with siblings as collateral informants. Mediation analysis showed that the effect of parental alcoholism on several adult outcomes was indirect, mediated by the other stresses and resources examined. Contextual models such as those presented here are helpful in understanding the long-term effects of childhood environment on women. PMID- 15804875 TI - Strategies for quitting among non-treatment-seeking marijuana smokers. AB - This study examines self-reported quitting strategies used by adult, non treatment-seeking marijuana smokers. Sixty-five subjects rated the use and effectiveness of thirteen strategies on a self-developed instrument, the Marijuana Quit Questionnaire. The strategies clustered into three categories/factors, whether grouped by principal components analysis, mean helpfulness rating, or frequency of endorsement: Change Environment, Seeking Organized/Professional Help, and Social Support. Changing one's environment was rated as most helpful while seeking help from professionals was the least helpful. Clinicians are likely to see marijuana users in their practice and should be proactive in offering assistance, incorporating the strategies reported here into treatment plans for their marijuana-using patients. PMID- 15804876 TI - Medical symptoms associated with tobacco smoking with and without marijuana abuse among crack cocaine-dependent patients. AB - Despite the widespread use of tobacco and marijuana by cocaine abusers, it remains unclear whether combined tobacco and marijuana smoking is more harmful than tobacco smoking alone in cocaine abusers. We investigated the differences in medical symptoms reported among 34 crack cocaine abusers who did not smoke tobacco or marijuana (C), 86 crack cocaine abusers who also smoked tobacco (C + T), and 48 crack abusers who smoked both tobacco and marijuana (C + T + M). Medical symptoms were recorded using a 134-item self-report instrument (MILCOM), and drug use was assessed using the Addiction Severity Index (ASI). After controlling for clinical and demographic differences, the C + T + M group reported significantly more total symptoms on the MILCOM as well as on the respiratory, digestive, general, and nose/throat subscales than the C + T or C groups. The C + T group reported higher total and respiratory and nose/throat symptoms than the C group. HOwever, the C group had the highest number of mood symptoms among the three groups. The C + T and C + T + M groups were comparable in number of cigarettes smoked and ASI scores. Although tobacco smoking is associated with higher reports of medical problems in crack abusers, smoking both marijuana and tobacco seems to be associated with greater medical problems than smoking tobacco alone. Tobacco smoking was not related to changes in cocaine use. Also, marijuana smoking does not appear to be associated with a reduction in tobacco or cocaine use. PMID- 15804877 TI - A cross-study comparison of cannabis and tobacco withdrawal. AB - A valid cannabis withdrawal syndrome has recently been established, but its clinical importance remains unclear. One method to assess the importance of cannabis withdrawal is to compare it with an established withdrawal syndrome. Cannabis and tobacco withdrawal studies that employed similar methods were used to compare six participant-rated and four observer-rated symptoms. Descriptive and graphic comparisons indicate that the magnitude and time course of withdrawal effects are similar across the two syndromes. These findings are consistent with other evidence supporting the clinical importance of the cannabis withdrawal syndrome. There remains a need for prospective experimental studies to replicate these findings. PMID- 15804879 TI - DSM-IV diagnostic criteria for pathological gambling: reliability, validity, and classification accuracy. AB - The purpose of this study was to examine the reliability, validity, and classification accuracy of the DSM-IV diagnostic criteria for pathological gambling. Given the lack of a laboratory test to diagnose pathological gambling, two groups were recruited in order to test DSM-IV diagnostic classification accuracy, one which likely had the disorder and the other which likely did not have the disorder (121 men and women clients at a gambling treatment facility) (138 men and women selected at random from the Windsor, Ontario, community who had gambled in the past twelve months). The Gambling Behavior Interview was administered to both groups. The Gambling Behavior Interview was administered to both groups. The Gambling Behavior Interview includes items that measure the ten DSM-IV diagnostic criteria for pathological gambling as well as other gambling problem severity measures and scales that served as tests of convergent validity. The ten DSM-IV diagnostic criteria were found to exhibit satisfactory reliability, validity, and classification accuracy; however, lowering the cut score to four and using item weights yielded improved classification accuracy over the standard cut score of five. Some diagnostic criteria were found to have greater discriminatory power than other criteria. The results of this study suggest that the classification accuracy of DSM-IV diagnostic criteria can be improved upon with a lower cut score or using weighted criteria. PMID- 15804878 TI - Lack of hippocampal volume change in long-term heavy cannabis users. AB - The effects of cannabis smoking on the morphology of the hippocampus are still unclear, especially because previous human studies have examined primarily younger, shorter-term users. We used magnetic resonance imaging to investigate these effects in a group of 22 older, long-term cannabis users (reporting a mean [SD] of 20,100 [13,900] lifetime episodes of smoking) and 26 comparison subjects with no history of cannabis abuse or dependence. When compared to control subjects, smokers displayed no significant adjusted differences in volumes of gray matter, white matter, cerebrospinal fluid, or left and right hippocampus. Moreover, hippocampal volume in cannabis users was not associated with age of onset of use not total lifetime episodes of use. These findings are consistent with recent literature suggesting that cannabis use is not associated with structural changes within the brain as a whole or the hippocampus in particular. PMID- 15804880 TI - Ego function as a correlate of addiction. AB - Eighty-nine substance abusers in an urban treatment facility completed the self report version of the Ego Function Assessment as well as the Addiction Severity Index. Statistical analyses delineate ego functions that relate to specific disruptions in the social and psychological adaptation of the substance abuser. Distinct profiles of ego functions are highlighted based on patterns of intravenous drug injection. Education levels are showen to correlate with ego function strengths. Results are interpreted from the social perspective of drug abuse. PMID- 15804902 TI - Communicating risk: from crisis to calm. PMID- 15804903 TI - "If we didn't use humor, we'd cry": humorous coping communication in health care settings. AB - Folkman and Lazarus's transactional theory of emotion and coping was used to explain how humor influences job satisfaction among health care providers. One hundred forty-two nurses completed measures of humor orientation (HO), coping efficacy, job satisfaction, and open-ended questions about their use of humor to relieve job tensions. This produced 9 categories of producing humor and 8 types of work situations identified in which humor was used to cope. Nurses (21.4%) reported using humor the most often during "patient care" situations (e.g., providing medicines, moving patients, physical therapy, and so on). More than one third (38.66%) of the nurses reported using "word-play/language" as a humorous coping strategy. Higher HO was associated with higher ratings of humor effectiveness, greater self-perceived coping efficacy, and higher emotional expressivity. Path analysis demonstrated that, as the transactional theory would predict, trait HO influences job satisfaction through its effect on heightened coping efficacy. Study limitations include the use of self-report methods and the limited number of responses to the open-ended items. Subsequent research in this area should attempt to validate the categories identified in this study and determine the most effective means of coping. Researchers also should explore differences in health care providers' coping communication based on gender, years of experience, and profession. PMID- 15804904 TI - The effect of antismoking advertisement executional characteristics on youth comprehension, appraisal, recall, and engagement. AB - This article examines how two executional characteristics of antismoking advertising may interact with other relevant advertising features to affect youth comprehension, appraisal, recall of, and engagement with antismoking ads. Fifty antismoking ads made by tobacco control agencies, tobacco companies, and pharmaceutical companies were appraised by 268 youth using an audience response methodology with a follow-up component. Analyses show that thematic and executional characteristics varied both across and within ad sponsor, and that executional characteristics of "personal testimonial" and "visceral negative" clearly had the strongest and most consistent effect on appraisal, recall, and level of engagement. Antismoking advertisements are not alike in their ability to engage youth. Advocates attempting to develop increasingly successful antismoking campaigns should consider the executional characteristics of proposed ads. PMID- 15804905 TI - HIV/AIDS risk factors as portrayed in mass media targeting First Nations, Metis, and Inuit peoples of Canada. AB - PURPOSE: The purpose of this study was to describe the coverage and portrayal of human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) risk factors as framed in newspaper targeting Aboriginal (First Nations, Metis, and Inuit) peoples in canada. METHODS: From a sample of 31 Aboriginal newspapers published in English from 1996 to 2000, 14 newspapers were randomly selected. Of 167 articles published on HIV/AIDS during this time period, all anecdotal (n=34) and an approximate 25% random sample of scientific (n=32) articles were analyzed using both quantitative (coding reliability and frequencies) and qualitative (in depth content analysis) analyses. RESULTS: Individual risk factors for HIV/AIDS were described in 74%, (49/66) of the articles and included unprotected sexual intercourse (20/49 or 41%), sharing of needles for injection drug use (IDU; 16/49 or 33%), infected blood transfusions (3/49 or 6%), and vertical transmission from mother to a baby (10/49 or 20%). Additional risk factors of alcohol use and poverty were mentioned in 29% and 25% of the articles. In addition to the well recognized HIV/AIDS risk groups of prostitutes and homosexual men, sexual abuse victims, prisoners, and women were identified in aboriginal newspapers as being at risk. Although Aboriginal women were identified as being at high risk, the newspaper coverage also emphasized their lack of knowledge regarding HIV/AIDS. Heterosexual men were not mentioned as being at risk for HIV/AIDS in the newspaper articles. BACKGROUND: The prevalence of HIV/AIDS is higher among Canadian Aboriginals than in the general population. Local and community newspapers are an important channels for the dissemination of health information for isolated, rural, and aboriginal communities. INTERPRETATION: The findings show that Aboriginal media identify high-risk groups and individualistic risk factors for HIV/AIDS within a public health perspective. PMID- 15804906 TI - Using research to inform health policy: barriers and strategies in developing countries. AB - This article examines the dissemination and uptake of health research into policy and program delivery in four developing countries. In-depth interviews were conducted with health researchers, policymakers, and practitioners at both the local and national level. The study highlights the similarities across the study countries in the barriers to effective dissemination and uptake of research results. A fundamental barrier to the uptake of research by decisionmakers is the lack of appreciation of the important contribution that research can make to policy and program development. A further barrier is researchers' lack of appropriate "packaging" of research findings that consider the needs of different policy audiences. Dissemination within academic circles also restricts access by decisionmakers nd practitioners. Overcoming the barriers requires effort on behalf of researchers, decisionmakers, and donor agencies. The strong presence of donor agencies in developing countries places them in a position both to enable and encourage dissemination activities and communication between researchers and policymakers or practitioners. Increased collaboration between all three parties is one of the key strategies toward increasing the uptake of research into health policy and program development. PMID- 15804907 TI - Evaluating the truth brand. AB - The American Legacy Foundation developed the truth campaign, an aspirational antismoking brand for adolescents. This study tested whether a multidimensional scale, brand equity in truth, mediates the relationship between campaign exposure and youth smoking. We collected brand equity responses from 2,306 youth on a nationally representative telephone survey. Factor analysis indicates that the scale has excellent psychometric properties and effectively measures brand equity. We developed a structural equation model to test the mediation hypothesis. Results show that brand equity mediates the relationship between truth and smoking. Analyses of potential cofounders show this relationship is robust. Behavioral branding (brands about a behavior or a lifestyle) is an important public health strategy. PMID- 15804909 TI - Announcement from the editor. PMID- 15804911 TI - Negotiating risk: how women working in massage parlours preserve their sexual and psychological health. AB - The small exploratory study was conducted in a heavily populated urban area in the Midlands region of England. Through semistructured interviews with women working in massage parlours, we investigated the health risks inherent in the work and the strategies adopted by the women to overcome these, as well as examining circumstances where theses strategies could be compromised. The effects of working in the sex industry on women's relationships and lives outside also emerged as the research progressed. We found that, in addition to preserving their sexual health, women's main concern was with the separation of home and work. PMID- 15804912 TI - Interviews with senegalese commercial sex trade workers and implications for social programming. AB - The purpose of the study was to assess the efficacy of the Senegalese pubic policy toward registered sex workers through an interview process examining their backgrounds and evaluating their knowledge of sexual health. Sixty registered sex workers in Dakar, Senegal, were interviews at the Institute d'Hygiene Social (IHS) to investigate patient knowledge of contraceptives and sexually transmitted infections (STIs). Ninety-eight percent of the women reported that, as a result of their visits to the clinic, they had increased precaution in their trade by demanding their clients use condoms and refusing clients who did not comply. Nearly 96% of the women were able to define the three main ways by which HIV is contracted, while 100% of the women reported that they used male condoms with their clients and would refuse clients who rejected the use of condoms. Senegal's proactive policy toward the safeguarding of women's health and the containment of HIV/AIDS through the legalization and monitoring of sex workers can serve as an example for successful strategies in the fight against the global spread of HIV/AIDS. PMID- 15804913 TI - HIV-related risk factors associated with commercial sex among female migrants in China. AB - Data from 633 sexually experienced female migrants were analyzed to examine the sociodemographic and psychosocial factors and human immunodeficiency virus (HIV) related behaviors associated with involvement in commercial sex. Six percent (40/633) of the participants reported having had sex for money. Compared with women who had not engaged in commercial sex, women who had sold sex were younger, less educated, and more likely to be unmarried. They were more likely to have engaged in HIV-related risk behaviors, such as becoming intoxicated with alcohol and using drugs. Among women engaged in commercial sex, only 28% of them consistently used condoms during the last three episodes of sexual intercourse. Women who had ever engaged in commercial sex demonstrated greater depressive symptoms than those without such as history (p < .01). Female migrants, especially those engaging in commercial sex, were vulnerable to HIV/sexually transmitted diseases (STDs). Sexual risk reduction and condom promotion are urgently needed among this population. Further studies are needed to examine the casual relationship between depression and HIV risk behaviors. PMID- 15804914 TI - Women's rights and health: the case of Oromo women in Ethiopia. AB - Human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS), for which we have no cure or vaccination, is the major health problem in Ethiopia. This epidemic generally has affected poor communities and women. To contain this epidemic, the empowerment of women is essential. In fact, the current working definition of health by the World Health Organization (WHO) makes social well being a part of everyday living, which is an essential dimension of the quality of life. The concept of quality of life means an opportunity to make choices and even change the situation one is in. Here, the concepts of health and human rights intersect, because of the quality of life requires freedom of choice, dignity and respect. Dignity, the right to access basic education and information, as well as the right to life are the major elements of human rights. It is for that purpose that health promotion is intended to enable people to increase their control over determinants of health and thereby improve their health. Women's involvement in outlining the agendas of health promotion is vital to achieve the desired goals. In this article I examine human rights violations in Ethiopia and the lost opportunities for Oromo women to make choices in life within the framework of the Universal Declarations of Human Rights (UDHR). PMID- 15804915 TI - "Becoming the real me": recovering from anorexia nervosa. AB - We investigated the process of recovery from anorexia nervosa using grounded theory. Open-ended interviews conducted with 9 women who had recovered from anorexia nervosa revealed that recovery focused on rediscovery and redefinition of the self and included the following: (1) seeing the dangers, (2) inching out of anorexia, (3) tolerating exposure without anorexia, (4) gaining perspective by changing the anorexia mindset, and (5) discovering and reclaiming self as "good enough." The process of recovery described in this study may provide a useful framework for helping women understand their own process of recovery efforts. The findings also support therapies that foster therapeutic alliance, acknowledge readiness for change, and promote a senses of autonomy. PMID- 15804916 TI - Letter to the editor. PMID- 15804917 TI - Introduction: mild cognitive impairment--cognitive, behavioral, and biological factors. PMID- 15804918 TI - The apolipoprotein E epsilon4 allele and incident Alzheimer's disease in persons with mild cognitive impairment. AB - Possession of one or more copies of the apolipoprotein E (APOE) epsilon4 allele is a known risk factor for Alzheimer's disease (AD), but it is uncertain whether the epsilon4 allele is associated with disease incidence among persons with mild cognitive impairment (MCI). We addressed this issue with data from the Religious Orders Study. Participants were 181 older Catholic clergy members who met criteria for MCI based on a uniform structured clinical evaluation; 56 (30.9%) had at least one epsilon4 allele. Clinical evaluations, which included clinical classification of dementia and AD, were repeated annually. During a mean of 5.7 years of observation, 79 persons (43.6%) developed AD. In a proportional hazards model that controlled for age, sex, and education, possession of an epsilon4 allele was associated with a 93% increase in the risk of developing Alzheimer's disease (95% CI; 1.02, 2.63). There was a marginally significant reduction in the effect of epsilon4 in older compared to younger participants (p=.053). The results suggest that possession of an epsilon4 allele does increase risk of AD in persons with MCI. PMID- 15804919 TI - Biomarker profiles and their relation to clinical variables in mild cognitive impairment. AB - The aim of the study was to compare clinical variables between MCI patients at different risk for Alzheimer's disease (AD) according to their biomarker profile. Fifty-four percent out of 39 MCI patients had a low Abeta42 and high tau in cerebrospinal fluid (CSF) (high-risk), 26% either a low CSF Abeta32 or high CSF tau (intermediate-risk) and 20% a normal CSF Abeta42 and tau (low-risk). Both high-and intermediate-risk subjects differed from the low-risk group in episodic memory, executive functions and the preclinical AD scale (PAS),which combines a set of clinical parameters. Subjects at high risk did not differ from subjects with an intermediate risk. Abeta42 levels correlated with the MTA and PAS scores, tau levels with episodic memory. These correlations suggest that the biomarkers are not independent when compared to the other AD markers. Longitudinal studies are necessary to interpret the correlations between biomarkers, imaging, and neuropsychological markers. PMID- 15804920 TI - FDG-PET measurement is more accurate than neuropsychological assessments to predict global cognitive deterioration in patients with mild cognitive impairment. AB - The accurate prediction, at a pre-dementia stage of Alzheimer's disease (AD), of the subsequent clinical evolution of patients would be a major breakthrough from both therapeutic and research standpoints. Amnestic mild cognitive impairment (MCI) is presently the most common reference to address the pre-dementia stage of AD. However, previous longitudinal studies on patients with MCI assessing neuropsychological and PET markers of future conversion to AD are sparse and yield discrepant findings, while a comprehensive comparison of the relative accuracy of these two categories of measure is still lacking. In the present study, we assessed the global cognitive decline as measured by the Mattis scale in 18 patients with amnestic MCI over an 18-month follow-up period, studying which subtest of this scale showed significant deterioration over time. Using baseline measurements from neuropsychological evaluation of memory and PET, we then assessed significant markers of global cognitive change, that is, percent annual change in the Mattis scale total score, and searched for the best predictor of this global cognitive decline. Altogether, our results revealed significant decline over the 18-month follow-up period in the total score and the verbal initiation and memory-recall subscores of the Mattis scale. The percent annual change in the total Mattis score significantly correlated with age and baseline performances in delayed episodic memory recall as well as semantic autobiographical and category word fluencies. Regarding functional imaging, significant correlations were also found with baseline PET values in the right temporo-parietal and medial frontal areas. Age and right temporo-parietal PET values were the most significant predictors of subsequent global cognitive decline, and the only ones to survive stepwise regression analyses. Our findings are consistent with previous works showing predominant delayed recall and semantic memory impairment at a pre-dementia stage of AD, as well as early metabolic defects in the temporo-parietal associative cortex. However, they suggest that only the latter predictor is specifically and accurately associated with subsequent cognitive decline in patients with MCI within 18 months of first assessment. PMID- 15804921 TI - Knight's move thinking? Mild cognitive impairment in a chess player. AB - We report the case of a chess player with superior premorbid cognitive function who presented to the Cognitive Disorders clinic at the National Hospital for Neurology and Neurosurgery with a 2-year history of symptoms of possible memory loss. Initially the MRI scan appearance was within normal limits and his cognitive scores inside the normal range; subsequently his cognitive function deteriorated and he fulfilled criteria for Mild Cognitive Impairment (MCI) two years later. Unexpectedly he died of an unrelated illness seven months later and post mortem examination of the brain was carried out, revealing advanced Alzheimer's disease (CERAD definite and NIA-Regan Institute high likelihood). This case highlights the difficulties encountered in assessing patients with superior premorbid function in the early stages of Alzheimer's disease, and reveals the value of serial MRI and neuropsychological assessment in detecting and monitoring early neurodegenerative disease. PMID- 15804922 TI - Cerebrospinal fluid TAU protein and amyloid beta42 in mild cognitive impairment: prediction of progression to Alzheimer's disease and correlation with the neuropsychological examination. AB - Cerebrospinal fluid (CSF) TAU protein and Amyloid beta42 were able to distinguish between 28 mild cognitive impairment (MCI) patients and both 38 normal aged and 17 anxious and depressed elderly patients, with good sensitivity/specificity when the two measures were combined. These biological markers are independent predictors of the presence of Alzheimer disease (AD), in addition to memory performance. Low Amyloid beta42 level was predictor of a fast progression of MCI patients to full blown dementia. The TAU protein level tended to correlate with memory performance, presumably in relation with the extent of the bilateral medio temporal damage in early AD. PMID- 15804923 TI - Mild cognitive impairment and everyday functioning in older adults. AB - The relations between mild cognitive impairment without dementia (MCI/CIND) and everyday functional abilities were examined using data from the Canadian Study of Health and Aging (CSHA). Individuals were identified with MCI/CIND if both caregiver report and clinician judgment agreed on the presence of cognitive impairment in the absence of dementia. Cross-sectional and longitudinal comparisons indicated that individuals with MCI/CIND demonstrated a broad range of impairment in instrumental activites of daily living (IADL) compared to individuals with no cognitive impairment (NCI). In cross-sectional analyses, neuropsychological measures of memory and psychomotor speed were significantly related to impairment in eight areas of functioning. In addition, poorer memory performance was significantly predictive of future impairment in money management. PMID- 15804924 TI - Years of severe and isolated amnesia can precede the development of dementia in early-onset Alzheimer's disease. AB - A young patient with a severe and isolated memory disorder, meeting the criteria for MCI, was followed for a period of 8 years. His overall functional level remained stable with a CDR-score at 0.5 until the last year when it dropped to 1.0. Neuropsychological testing showed severe memory deficits but otherwise normal cognitive functions. Only minimal progression was measured; however, the last testing showed impaired abstraction and executive functioning as well as discrete problems generating names of objects and people. Neuroimaging, with MRI and SPECT, was consistent with a progressive degenerative disorder, and cerebrospinal fluid levels of beta-amyloid 1-42, tau protein, and phosphorylated tau protein were characteristic of Alzheimer's disease (AD). We argue that this is a case of prodromal AD, which illustrates an extreme version of the normal course with respect to slow progression of the disease and severity of amnesia early in the course. PMID- 15804925 TI - A decade of pre-diagnostic assessment in a case of familial Alzheimer's disease: tracking progression from asymptomatic to MCI and dementia. AB - Detailed study of the very earliest phases of Alzheimer's disease (AD) is seldom possible, especially those changes preceding the development of mild cognitive impairment (MCI), which may occur years before diagnosis. Knowledge of imaging and neuropsychological features of these early stages would add insight into this poorly understood phase of the disease. We present data from a subject who entered a longitudinal study of individuals at risk of familial Alzheimer's disease (FAD), as a healthy volunteer with no memory complaints, undergoing 12 assessments between 1992 and 2003. Longitudinal MRI, neuropsychological and clinical data are presented over the decade preceding this man's diagnosis, through the asymptomatic and prodromal preludes to his presentation with MCI and on to eventual conversion to AD. PMID- 15804926 TI - Non-cognitive symptoms in mild cognitive impairment subjects. AB - The term mild cognitive impairment (MCI) is used to identify individuals with worse cognitive performance than those with normal aging, and who are at risk of dementia, especially Alzheimer's disease (AD). Although the MCI concept is based on the presence of specific cognitive deficits, several studies have shown that these subjects can develop depression, disruptive behaviors (e.g., agitation, aggression), and psychosis. In this study, we examined the baseline psychiatric characteristics of 228 MCI (Mean age: 71.2, Mini-mental State Examination [MMSE] score: 25.9) and 427 mildly demented Probable AD (Mean age: 73.2; Mean MMSE score: 23.5) subjects from a referral dementia clinic. The psychiatric assessment was conducted by geriatric psychiatrists using semi-structured interviews. The proportion of subjects with major depression (MCI: 7.5% vs. Probable AD: 8%) and aggression (MCI: 10% vs. Probable AD: 12.5%) was similar in the two groups. There were more Probable AD patients with psychomotor agitation (52% vs. 38%), delusions (29% vs. 14%), and hallucinations (9% vs. 4%) than MCI subjects. Within MCI groups, we did not observe any differences between MCI subjects with amnesic syndrome versus MCI subjects with a much broader cognitive deficit. These results showed that the MCI syndrome is not circumscribed to a neuropsychological definition, but occurs with a wide range of psychiatric syndromes. Furthermore, it is possible that the development of disruptive behaviors and psychosis, in MCI subjects with no previous history of psychiatric illness, constitutes a strong indication that there is an underlying neurodegenerative disorder. PMID- 15804928 TI - Abnormal visual search in mild cognitive impairment and Alzheimer's disease. AB - Our aim was to further characterize the clinical concept of mild cognitive impairment (MCI). We examined the status of visual attention-related processing in such patients in relation to healthy older adults and patients with Alzheimer's disease (AD) by measuring performance on a computer-based visual search task. We tested 20 older adult control participants, 13 patients with amnestic mild cognitive impairment and 12 patients with AD. Patients with AD and with MCI exhibited a significant detriment in visual search performance compared to the older adult controls. The deficit in visual search was greater for the patients with AD than the patients with MCI. The pattern of results displayed by the MCI group indicates that patients who appear clinically to suffer only from a deficit in memory also display a deficit in visual attention-related processing, which although not as severe as those with AD, represents a significant detriment in such performance compared to that seen in healthy ageing. PMID- 15804927 TI - Stability and predictability of the classification of mild cognitive impairment as assessed by episodic memory test performance over time. AB - This study aimed to address the criteria and the stability of the classification of MCI. The Foresight Challenge cohort of 157 community-dwelling volunteers was assessed on 3 visits at 2-year intervals with episodic, semantic and working memory tests. Subjective memory complaints were assessed with the CAMDEX. Of the cohort, 2% had dementia and 31% were classified with MCI at visit 3, 43% with stable impairment from Time 1. Thirteen percent of those with objective memory impairment at Time 1 or 2 improved to control status by Time 3. Episodic memory tests were predictive for MCI at all timepoints, as were tests for praxis and Graded Naming, while at Time 3 spatial span lost predictive value, but processing speed became predictive. Decline in processing speed was seen in control and MCI groups, while memory performance and MMSE decline occurred only in the MCI group. The use of combined memory test scores gave better sensitivity to MCI than single tests. Subjective memory complaints were positive for 79% of the MCI group and 62.5% of controls. These findings would suggest consideration of modification of current MCI criteria. PMID- 15804929 TI - Abnormal spatial and non-spatial cueing effects in mild cognitive impairment and Alzheimer's disease. AB - Our aim was to further characterize the clinical concept of mild cognitive impairment (MCI). We examined visual attention-related processing in 12 patients with amnestic MCI, 16 healthy older adults and 16 patients with Alzheimer's disease (AD) by measuring performance on computer-based tests of attentional disengagement, alerting ability, and inhibition of return. Unlike the healthy older controls, the patients with AD and the patients with amnestic MCI exhibited a significant detriment in both the ability to disengage attention from an incorrectly cued location and the ability to use a visual cue to produce an alerting effect. The pattern of results displayed by the MCI group indicates that patients who only appear clinically to suffer from a deficit in memory also display a deficit in specific aspects of visual attention-related processing, which closely resemble the magnitude seen in AD. PMID- 15804930 TI - Healing the disembodied mind: contemporary models of conversion disorder. AB - The cartesian separation of body and mind has underpinned medical conceptualization of conversion symptoms over the last four centuries. Eighteenth and nineteenth-century models explained conversion symptoms on the basis of assumed organic lesions. Subsequent psychological formulations focused on mental phenomena, which were disembodied from underlying neurobiological processes. Contemporary theories challenge this disconnection. They place conversion symptoms within an evolutionary framework, arguing that mind is embodied in physical processes and confers significant survival advantage. Current neurobiological models explain conversation symptoms as part of the human emotional response to threat, reflecting either errors in how information about body state is processes and represented, or the motor component of an automatic emotional response. Patients with conversion disorder experience their body functions in a range of distressing and disturbing ways. In order to understand these phenomena, clinicians require conceptual models that expand the conventional dualistic mind-body paradigm, integrate multiple theoretical perspectives, and take into account environmental factors that shape evolutionary and individual development. PMID- 15804932 TI - Toward convergence in the medication treatment of bipolar disorder and schizophrenia. AB - Reaching a correct differential diagnosis among patients with psychotic symptoms was especially important during the era of first-generation antipsychotics, when treatments for the different disorders varied in terms of adverse events and likelihood of response. The historical "overdiagnosis" of schizophrenia and "underdiagnosis" of bipolar disorder in the United States was blamed for an increased exposure to neuroleptics among patients who might have benefited from lithium. With the recognition that second-generation antipsychotics are useful in the treatment of both schizophrenia and bipolar mania, and that combining them with classic mood stabilizers such as valproate may results in increased efficacy, the field is witnessing a convergence of pharmacological approaches to the treatment of schizophrenia and bipolar disorder. Substantially more data is available regarding combination treatments for bipolar disorder than for schizophrenia, and appropriate diagnosis remains important in predicting prognosis, but until the precise pathophysiology of psychotic disorders can be elucidated, and specific targeted treatments crafted, we will continue to see similar blended treatments for these two disease states. PMID- 15804933 TI - Acting (to) out: approaching homosexuality in a 15-year-old boy with anxiety and depression. PMID- 15804931 TI - Temperament and its role in developmental psychopathology. AB - Temperament refers to early-appearing variation in emotional reactivity. The core dimensions of temperament and optimal method for assessment continue to be sources of considerable discussion. Nevertheless, the moderate stability of most temperamental traits and the strong influence of genetic and unique environmental factors have been well established, along with temperament's association with childhood psychiatric disorders. Both temperamental predisposition toward experiencing negative emotions and low inhibitory control are linked to many psychiatric conditions, while other dimensions, such as levels of extraversion, vary by, and likely even within, disorders. Accumulating research directed at understanding the mechanism of these links between temperament and psychopathology indicate that, at least for most disorders, the two constructs cannot be viewed as simply different points along a shared continuum. The effect of temperament upon psychopathology has been found to be mediated and moderated by a number of both internal and external factors. Additional research is needed to help further define the core dimensions of temperament and the complex mechanisms through which temperamental traits interact with other influences in affecting developmental trajectories. PMID- 15804934 TI - Education designing a sleep disorders curriculum for psychiatry residents. PMID- 15804935 TI - Effects of subchronic exposures to concentrated ambient particles (CAPs) in mice. I. Introduction, objectives, and experimental plan. AB - This subchronic (6-mo) inhalation study of the effects of concentrated ambient air fine particulate matter (PM2.5) in normal mice (C57) and a murine model of humans with an advanced level of aortic plaque (ApoE-/- or ApoE-/- LDLr-/-) was designed to determine the presence and extent of a variety of health-related responses. The animals were exposed for 6 h/day, 5 day/wk during the spring and summer of 2003 to concentrations that were elevated 10-fold in Tuxedo, NY, a regional background site that is upwind and approximately 50 km west-northwest of New York City. The average PM2.5 concentration during exposure was 110 microgram/m3, and the long-term average was 19.7 microg/m3. There were substantial daily variations in concentration, and we sought evidence both for the influence of peak exposures on acute responses and for the cumulative effects of the prolonged series of exposures. Acute responses were characterized in terms of: (1) short-term electrocardiographic (EKG), core body temperature, and physical activity differences between PM and sham-exposed mice; and (2) in vitro toxicity of a simultaneously collected PM2.5 sample to lung epithelial cells. Cumulative responses to PM2.5 were characterized in terms of changes in heart rate, heart-rate variability, heart-rate variance, aortic plaque density, genetic marker expression, and brain cell distributions. There were no significant changes in the normal mice. The nature and extent of the exposure-related responses that were seen in the ApoE-/- as well as ApoE-/- LDLr-/- mice are described in the articles that follow in this special issue of Inhalation Toxicology. PMID- 15804936 TI - Effects of subchronic exposures to concentrated ambient particles (CAPs) in mice. II. The design of a CAPs exposure system for biometric telemetry monitoring. AB - We modified, assembled, tested, and validated the versatile aerosol concentration enrichment system (VACES) developed by Sioutas et al. (1999) for use in a subchronic experiment that involved exposure of mice in vivo and of respiratory epithelial cells in vitro to concentrated ambient particles (CAPs). Since the labor-intensive nose-only exposure regimen is not an option in a long-term experiment, a whole-body exposure mouse chamber was designed specifically for use with the VACES. The exposure system concsists of a stainless-steel (SS) tub with 32 cubicles (1 mouse per cubicle) separated by perforated SS sheets. The tops of these cubicles are covered with perforated plastic sheets to allow telemetry monitoring during the exposure. In each exposure chamber, perforated aluminum tubes are used to distribute CAPs evenly (within 2% difference) throughout the exposure chamber. The exhaust consists of perforated aluminum tubes covered with a urine shield. The modification to the original design of the VACES facilitated the operation of the system in a subchronic study. Mass flow controllers maintain a constant flow rate into the exposure chambers. For a sham control exposure, the identical system is used, except that a HEPA filter at the inlet to the VACES removes 98% of ambient particles. The entire system allow for simultaneous exposure of 64 mice to CAPs, with an equal number of sham-expose mice as controls. Telemetry receives have been modified so that 16 mice per group with electrocardiograph (EKG) transmitters can be monitored during exposure. Furthermore, a BioSampler is used to collect CAPs (one sample per day) for the in vitro exposures. In this article, the assessments of flow and particle distribution of the exposure chamber as well as the performance of the system during the subchronic exposure experiment are described. PMID- 15804937 TI - Effects of subchronic exposures to concentrated ambient particles (CAPs) in mice. III. Acute and chronic effects of CAPs on heart rate, heart-rate fluctuation, and body temperature. AB - Normal mice (C57) and mice prone to develop atherosclerosis (ApoE-/-) were implanted with electrocardiograph (EKG), core body temperature, and motion transmitters were exposed daily for 6 h to Tuxedo, NY, concentrated ambient particles (CAPs) for 5 day/wk during the spring and summer of 2003. The series of 5-min EKG monitoring and body-temperature measurements were obtained for each animal in the CAPs and filtered air sham exposure groups. Our hypothesis was that chronic exposure could cause cumulative health effects. We used our recently developed nonparametric method to estimate the daily time periods that mean heart rates (HR), body temperature, and physical activity differed significantly between the CAPs and sham exposed group. CAPs exposure most affected heart rate between 1:30 a.m. and 4:30 a.m. With the response variables being the average heart rate, body temperature, and physical activity, we adopted a two-stage modeling approach to obtain the estimates of chronic and acute effects on the changes of these three response variables. In the first stage, a time-varying model estimated daily crude effects. In the second stage, the true means of the estimated crude effects were modeled with a polynominal function of time for chronic effects, a linear term of daily CAPs exposure concentrations for acute effects, and a random component for unknown noise. A Bayesian framework combined these two stages. There were significant decreasing patterns of HR, body temperature, and physical activity for the ApoE-/- mice over the 5 mo of CAPs exposure, with smaller and nonsignificant changes for the C57 mice. The chronic effect changes of the three response variables for ApoE-/- mice were maximal in the last few weeks. There was also a significant relationship between CAPs exposure concentration and short-term changes of heart rate in ApoE-/- mice during exposure. Response variables were also defined for examining fluctuations of 5-min heart rates within long (i.e., 3-6 h) and short time periods (i.e., approximately 15 min). The results for the ApoE-/- mice showed that heart-rate fluctuation within the longer periods increased to 1.35-fold by the end of exposure experiment, while the heart-rate fluctuation within 15 min decreased to 0.7-fold. PMID- 15804938 TI - Effects of subchronic exposures to concentrated ambient particles (CAPs) in mice. IV. Characterization of acute and chronic effects of ambient air fine particulate matter exposures on heart-rate variability. AB - Long-term exposure to fine particulate air pollution (PM2.5) has been associated increased risk of death from cardiopulmonary diseases. Cardiac function parameters have also been affected by ambient particulate matter (PM) exposure, including heart-rate variability (HRV), a measure of autonomic function that has been recognized as a well-defined, quantitative indicator of autonomic dysfunction. However, the role of HRV in ambient PM-induced cardiovascular effect is not fully understood. In an accompanying article, we report significant decreasing patterns of heart rate (HR), body temperature, and physical activity for mice lacking apoliproprotein (ApoE-/-) over 5 mo of exposure to concentrated ambient PM (CAPs), with smaller and nonsignificant change for C57 mice. In this article, we report the effects of subchronic CAPs exposure on HRV parameters that are sensitive to cardiac sympathetic and parasympathetic nerve activity. The standard deviation of normal to normal beat intervals (SDNN) and the square root of the mean squared differences of successive RR intervals (RMSSD) in the late afternoon and overnight for the ApoE-/- mice showed a gradual increase for the first 6 wk, a decline for about 12 more wk, and a slight turn upward at the end of the study period. For C57 mice, there were no chronic effect changes of SDNN or RMSSD in the late afternoon, an a slight increase after 6 wk for the overnight period. The response patterns of ApoE-/- mice indicated a perturbation of the homeostatic function in the cardiovascular system (initial enhancement and late depression of the HRV parameters). Our results complement the findings in human panel and controlled CAPs exposure studies in demonstrating that increased levels of particle pollution are able to perturb cardiac autonomic function, which may lead to adverse cardiovascular outcomes. PMID- 15804939 TI - Effects of subchronic exposures to concentrated ambient particles (CAPs) in mice. V. CAPs exacerbate aortic plaque development in hyperlipidemic mice. AB - Recent epidemiological studies suggest that long-term exposure to particulate matter (PM) causes chronic effects on the cardiovascular system that result in cumulative increases cardiovascular morbidity and mortality. Since atherosclerosis is a progressive irreversible condition and an underlying cause of many cardiovascular diseases, we hypothesized that long-term exposure to PM causes adverse cardiovascular effects by exacerbating atherosclerosis. In this study, we exposed C57- and ApoE-deficient (ApoE-/-) and ApoE, LDLr (DK)-deficient mice to concentrated ambient PM2.5 for 6 h/5 days/wk, for up to 5 mo. The overall mean exposure concentration for these groups of animals was 110 microg/m3. The cross-sectional area of the aorta root of DK mice was examined morphologically using confocal microscopy for the severity of lesion, extent of cellularity, and lipid contents. Aortas from the arch to the iliac bifurcations were also sectioned longitudinally and lesion areas were stained with Sudan IV. All DK mice regardless of exposure had developed extensive lesions in the aortic sinus regions, with lesion areas that covered more than 79% of the total area. In male DK mice, the lesion areas in the aortic sinus regions appeared to be enhanced by concentrated ambient particles (CAPs), with changes approaching statistical significance (p = .06). In addition, plaque cellularity was increased by 28% (p = .014, combined), whereas there were no CAPs-associated changes in the lipid content in these mice. When examining the entire aorta opened longitudinally, both the ApoE-/- and DK mice had prominent areas of severe atherosclerosis covering 40% or more of the lumenal surface. Visual examination of all images suggested that plaques tend to form in clusters concentrating near the aortic arch and and the iliac bifurcations. Quantitative measurements showed that CAPs exposure increased the percentage of aortic intimal surface covered by grossly discernible atherosclerotic lesion by 57% in the ApoE-/- mice (p = .03). Changes produced by CAPs in male (10% increase) or female DK mice (8% increase) were not statistically significant. In this study, we have demonstrated that subchronic exposure to CAPs in mice prone to develop atherosclerotic lesions had a significant impact on the size, severity, and composition of aortic plaque. PMID- 15804940 TI - Effects of subchronic exposures to concentrated ambient particles (CAPs) in mice. VI. Gene expression in heart and lung tissue. AB - The purpose of this exploratory study within the integrated subchronic inhalation exposure study (Lippmann et al., 2005) was to identify genes in heart and lung tissue that changed in expression level as a result of subchronic exposure to concentrated ambient particles (CAPs). Identification of CAPs exposure-related changes in gene expression could serve in the formulation of mechanistic hypotheses and/or to suggest possible biomarkers of exposure. In this exploratory study undertaken here, tissues from multiple replicates of ApoE/low-density lipoprotein double knockout (DK) mice were examined for relative exposure-related changes in gene expression. Due to limited resources, the number of replicates was three for each tissue (lung and heart) of each exposure condition (CAPs or air control). A rigorous comparison of exposure versus control data using the "significance analysis of microarrays" (SAM) method indicated that only one gene was differentially expressed at a significant level. However, when using a less restrictive, nonstatistical analytical treatment of the data, several genes that might be involved in PM-related heart or lung pathology, and/or the circadian rhythm of physiological processes, were identified. A more comprehensive study is required to mre definitively assess differences in gene expression in heart and lung resulting from exposure to CAPs. PMID- 15804941 TI - Effects of subchronic exposures to concentrated ambient particles. VII. Degeneration of dopaminergic neurons in Apo E-/- mice. AB - This study reports that subchronic exposure of Tuxedo, NY concentrated ambient particulates (CAPs) produces neuropathological damage in the brains of Apo E deficient mice (Apo E-/-). These genetically modified mice are characterized by elevated levels of oxidative stress (OS) in the brain. Microscopic examination of coronal sections of the brain, immunocytochemically stained for dopamineric neurons, indicated that neurons from the substantia nigral nucleus compacta were significantly reduced by 29% in CAPs-exposed Apo E-/- mice relative to air exposed Apo E-/- controls. In addition, statistically significant increases (p < .05) in immunocytochemically stained astrocytes were noted. The dopaminergic neurons of the nucleus compact are specifically targeted in Parkinson's disease. The present study expands the systems affected by particulate matter to include the brain, and supports an environmental role for the development of neurodegeneration in OS-susceptible individuals. PMID- 15804942 TI - Effects of subchronic exposures to concentrated ambient particles (CAPs) in mice. VIII. Source-related daily variations in in vitro responses to CAPs. AB - The NYU PM Center subchronic animal inhalation study addressed the issues of composition and sources of ambient fine particulate matter (PM2.5), as well as the relationship of these PM2.5 characteristics to the cellular response of human bronchial epithelial cells. In this simultaneous study, we used an in vitro exposure technique to compare the daily variations of the responses of cells to fine concentrated ambient particles (CAPs) collected from a rural area upwind of New York City for the period of 9 a.m. to 3 p.m. on weekdays only, March September, 2003. Chemical composition data for CAPs were modeled using factor analysis with varimax orthogonal rotation to determine four particle source categories contributing significant amount of mass to CAPs at Sterling Forest (Tuxedo, NY). These source categories are: (1) regional secondary sulfate characterized by high S, Si, and organic carbon (OC); (2) resuspended soil characterized by high concentrations of Ca, Fe, Al, and Si; (3) oil-fired power plants emissions of the eastern United States identified by presence of V, Ni, and Se; and (4) unknown other sources. To estimate the mass contributions of each individual source category, the CAPs mass concentration was regressed against the factor scores. Regional sulfate was the largest contributor to mass (65%), followed by soil (20%), residual oil combustion (2%), and the other sources contributing 13%. Based on an evaluation of the cellular responses to CAPs and a detailed chemical characterization of the ambient PM2.5, we investigated the sources and individual components of ambient PM2.5 that are responsible for the induced cellular response. Nuclear Factor kappa B (NF-kappa B) was selected as a monitor of cellular stress response that followed after the exposure to CAPs. The results of the NF-kappa B assay were found to be most highly correlated with Ni and V among the individual components, and with the residual oil combustion source category. PMID- 15804943 TI - Effects of subchronic exposures to concentrated ambient particles in mice. IX. Integral assessment and human health implications of subchronic exposures of mice to CAPs. AB - In order to examine the biologic plausibility of adverse chronic cardiopulmonary effects in humans associated with ambient particulate matter (PM) exposure, we exposed groups of normal mice (C57) and knockout mice that develop atherosclerotic plaque (ApoE-/- and ApoE-/- LDLr-/-) for 6 h/day, 5 days/wk for 5 or 6 mo during the spring/summer of 2003 to either filtered air or 10-fold concentrated ambient particles (CAPs) in Tuxedo, NY (average PM2.5 concentration during exposure = 110 microg/m3). Some of the mice had implanted electrocardiographic monitors. We demonstrated that: (1) this complex interdisciplinary study was technically feasible in terms of daily exposure, collection of air quality monitoring data, the collection, analysis, and interpretation of continuous data on cardiac function, and the collection and analyses of tissues of the animals sacrificed at the end of the study; (2) the daily variations in CAPs were significantly associated, in ApoE-/- mice, with daily variations in cardiac functions; (3) there were significant differences between CAPs and sham-exposed ApoE-/- mice in terms of cardiac function after the end of exposure period, as well as small differences in atherosclerotic plaque density, coronary artery disease, and cell density in the substantia nigra in the brain in the ApoE-/- mice; (4) there are suggestive indications of gene expression changes for genes associated with the control of circadian rhythm in the ApoE-/- LDLr-/- double knockout (DK) mice. These various CAPs-related effects on cardiac function and the development of histological evidence of increased risk of clinically significant disease at the end of exposures in animal models of atherosclerosis provide biological plausibility for the premature mortality associated with PM2.5 exposure in human subjects and provide suggestive evidence for neurogenic disease as well. PMID- 15804944 TI - You do not have to work that hard to be successful and entirely happy. PMID- 15804945 TI - William J Sneed: surgeon, humanist, and educator. PMID- 15804946 TI - The effect of interleukin-10 on acute pancreatitis induced by cerulein in a rat experimental model. AB - To investigate whether interleukin-10, a potent anti-inflammatory cytokine, could have a therapeutic effect on rats on that were made pancreatitis by cerulein. Thirty Wistar Albino rats were randomized into sham, pancreatitis, and therapy groups (n = 10 in each). Nothing was applied to the sham group; pancreatitis by inject-ing cerulein (50 micro/g/kg/h) was induced in the pancreatitis and therapy groups. Interleukin-10 (10.000 U) was injected at 1 and 4 h after pancreatitis inductions in the therapy group. The rats were sacrificed at postoperative hour 24. The following parameters were investigated: the leukocyte count, blood glucose, amylase, lipase and tumor necrosis factor-alpha levels in the blood samples; histopathological search, and wet/dry weight ratios of the pancreas tissues. The ratio of wet/dry pancreatic tissue weight, serum tumor necrosis factor-alpha, amylase and lipase lev-els, and histologic damage scores in the pancreatitis and therapy groups were significantly higher when they were compared with the sham group(p < .01). However, all of these values were significantly lower in the therapy groups than in the pancreatitis group (p < .01). Interleukin 10 decreases pancreatic tissue injury induced by cerulein-induced pancreatitis in rats. Nevertheless, more experimental studies are needed to compare endogenous interleukin-10 with exogenous interleukin-10 effects before clinical usage of this drug. PMID- 15804947 TI - Syngeneic islet transplantation into seminal vesicles of diabetic rats. AB - Pancreatic islet transplantation has been proposed as an attractive option for the treatment of type I diabetes. Transplantation into different sites has been investigated, among them those that are immuno-logically privileged (e.g., thymus, uterus, brain, anterior eye chamber, and testicle). Because of their characteristics, seminal vesicles could be considered as immunologically privileged organs, but there is no worldwide experience that can confirm it. The purpose of the present study is to assess the viability and functionality of islet transplantation into seminal vesicles of diabetic rats. One hundred ninety inbred adult male syngeneic Lewis rats were used as donors (n = 72), receptors (n = 36), and controls(n = 11). Diabetes was chemically induced through a single intraperitoneal injection of streptozotocin. Groups of 1200 purified islets were introduced in the right seminal vesicle of diabetic rats. Diabetic control rats were sham transplanted. Body weight and glycemia were monitored every 2 d. Of transplanted rats, 16.7% achieved a good function due to islet engraftment, while 30.6% achieved a partially good response, and 52.7% were considered as nonresponding. This is the first report about islet transplantation into seminal vesicles of diabetic animals. Our results indicate that islet transplantation into rat seminal vesicles is technically possible, and that islets can function normally after engraftment into the wall of the seminal vesicle. PMID- 15804948 TI - Passive biomechanical properties of sutured mammalian muscle lacerations. AB - Muscle trauma, such as laceration or transection, is a common occurrence, but repairing delicate tissue poses a clinical challenge. This is at least partially due to the lack of established muscle repair models. The purpose of this study was to compare the biomechanical properties of stitches in transected porcine and bovine muscle bellies. A biomechanical protocol was designed for measuring suture performance in muscle belly lacerations. Twenty simple stitches in porcine and 21 stitches in bovine specimens were tested. Individual stitches were placed in lacerated muscle bellies and tensioned on a biomechanical tester (model 8521S, Instron Corporation, Canton, MA). The mean maximum load for porcine (22.0 N) and bovine (23.9 N)stitches was not significantly different (p = .48). The difference in mean strains at maximum load between porcine (9.70/%) and bovine(8.0%0/) groups was statistically significant (p = .004). Failure mechanisms were similar. One porcine stitch avulsed the muscle transversely,while 19 stitches tore out longitudinally. All 20 stitches tore out in bovine specimens. Sutured muscle was the weakest element in each test. The present study demonstrated that sutured muscles performed similarly for the two mammals regarding the parameters of maximum load and mechanism of failure. Regarding suturing of skeletal muscle lacerations, both mammalian models had similar biomechanical performance for maximum loads and failure mechanisms, while strain data differed. The stitch load magnitudes in this study approximate those required to successfully repair transected muscle. Knowledge introduced by this study fills a gap concerning muscle stitching relevant to clinical care. PMID- 15804949 TI - 2-Arachidonoylglycerol increases in ischemia-reperfusion injury of the rat liver. AB - Several studies have implicated endocannabinoids in various forms of shock. However, the role of endocannabinoids in hepatic ischemia-reperfusion injury remains unclear. The purpose of this study was to evaluate the changes of two endocannabinoidsin hepatic ischemia-reperfusion injury: anandamide (ANA) and 2 arachidonoylglycerol (2-AG). Male Sprague-Dawley rats were divided into 2 groups: the short (15 min) ischemic group and the long (60 min)ischemic group in the segmental (70%) hepatic tissue. Blood levels of aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), ANA, and 2-AG were examined. Serum lev-els of AST, ALT, and LDH were significantly higher in the long-ischemia group than in the short-ischemia group. Plasma levels of 2-AG showed similar augmentation prior to and after reperfusion in both the short- and long-ischemia groups, although plasma 2-AG lev-els tended to be higher in the long-ischemia group than in the short-ischemia group. Plasma levels of ANA were augmented in the early phase of reperfusion in the short-ischemia group and did not differ significantly from the normal level with time after reperfusion in the long-ischemia group. These results suggest that the endocannabinoid 2-AG increases in hepatic ischemia-reperfusion injury of rats, rather than ANA. PMID- 15804950 TI - Reduced expression of collagen type I and increased expression of matrix metalloproteinases 1 in patients with Crohn's disease. AB - Crohn's disease (CD) is a chronic inflammatory bowel disease of still unknown etiology. The aim of our study was to find out whether there are any changes in the colonic wall of CD patients that could give hints for a predisposing disorder concerning the extracellular matrix, especially the collagen metabolism. Eight samples of colonic tissue from patients with Crohn's disease were compared to 14 specimens from patients without Crohn's disease. We performed a sirius red test for the overall collagen content and immunohistochemical studies examining differentiation between" collagen type I and type III and the expression of MMP-1 and MMP-13. In the bowel sections of patients with Crohn's disease, decreased levels of mature collagen type I with a resulting lower ratio of collagen I/III compared to patients without Crohn's disease were found (1.12 +/- 0.29 vs. 1.59 i 0.31). The expression of MMP-1 was significantly increased in the CD group (9.21 i 6.02 vs.6.02 i 1.98), whereas expression of MMP-13 showed no difference in both groups. Our study gives the first indication that preexisting changes of the extracellular matrix in the colonic wall may play a role in the pathogenesis of CD. Further studies have to be done to elucidate these interesting aspect of the pathogenesis in Crohn's disease. PMID- 15804951 TI - Preoperative risk factors associated with mortality in lung biopsy patients with interstitial lung disease. AB - The objective of this study was to evaluate the risk factors associated with mortality in interstitial lung disease patients. We performed a retrospective study of 722 consecutive patients submitted for lung biopsy during the 1986-1990 period. Twenty-two (3%) died within the 30 days following surgery. Forty-four patients who survived after the surgery for the same time span were randomly chosen as control group. Dyspnea at rest was present in 18/44 of surviving group (SG) and in 18/22 of the nonsurviving group (NSG) (OR 6.5, 95% CI 1.8-22.4,p = .001). Systemic diseases (i.e., diabetes, systemic arterial hypertension)were mainly present in the NSG (OR 7.2, 95% CI 2.3-22.8, p < .001). The SG displayed significantly less respiratory insufficiency with a PaO2 of 52.2 + 8.4 versus 38.5 i 9.4 mm Hg, and PaCO2 of 28.8 i 4.5 versus 38.5 +/- 9.2 mm Hg, respectively (p < .001). Likewise, the SG exhibited a PaCO2/PaO2 ratio of 0.5 - 0.1, while in the NSG it was of 1 +/- 0.4 (p < .001), showing a sensitivity of 84% and specificity of 93% for mortality. Multiple logistic regression analysis for these variables showed that log likelihood was still significant for PaCO2 > 34 mm Hg, PaO2 <48 mm Hg, and comorbid diseases. Logistic regression analysis of these three variables showed the greatest sensitivity and specificity (84 and 750/0,respectively) for prediction of mortality. However, the strongest association was found when PaCO2/PaO2 ratio was analyzed alone (OR 21,073,CI 95% 28-15,946,357, p < .005). These data suggest that PaCO2/PaO2 ratio appears to be a predictor of mortality in this subset of patients. Its prospective use has reduced early mortality after surgery less than 1% in the last decade. PMID- 15804952 TI - Acute hepatic failure in swine: hepatectomy vs. vascular occlusion. PMID- 15804954 TI - Autistic disorder and viral infections. AB - Autistic disorder (autism) is a behaviorally defined developmental disorder with a wide range of behaviors. Although the etiology of autism is unknown, data suggest that autism results from multiple etiologies with both genetic and environmental contributions, which may explain the spectrum of behaviors seen in this disorder. One proposed etiology for autism is viral infection very early in development. The mechanism, by which viral infection may lead to autism, be it through direct infection of the central nervous system (CNS), through infection elsewhere in the body acting as a trigger for disease in the CNS, through alteration of the immune response of the mother or offspring, or through a combination of these, is not yet known. Animal models in which early viral infection results in behavioral changes later in life include the influenza virus model in pregnant mice and the Borna disease virus model in newborn Lewis rats. Many studies over the years have presented evidence both for and against the association of autism with various viral infections. The best association to date has been made between congenital rubella and autism; however, members of the herpes virus family may also have a role in autism. Recently, controversy has arisen as to the involvement of measles virus and/or the measles, mumps, rubella (MMR) vaccine in the development of autism. Biological assays lend support to the association between measles virus or MMR and autism whereas epidemiologic studies show no association between MMR and autism. Further research is needed to clarify both the mechanisms whereby viral infection early in development may lead to autism and the possible involvement of the MMR vaccine in the development of autism. PMID- 15804955 TI - Characterization of West Nile viral replication and maturation in peripheral neurons in culture. AB - The North American West Nile virus (WNV), New York 1999 strain, appears to be highly neurotropic, and its neuroinvasiveness is an important aspect of human disease. The authors have developed an in vitro model to study WNV replication and protein processing in neurons. They compared WNV infection of the dorsal root ganglion (DRG) neurons (sensory neurons) and PC-12 cells (sympathetic neurons) to WNV infection of the mosquito cell line, C6/36, and Vero cells. WNV infection of both neuronal cell types and C6/36 cells was not cytopathic up to 30 days post infection, and continual viral shedding was observed during this period. However, WNV infection of Vero cells was lytic. Interestingly, WNV infection of neurons was not efficient, requiring a high multiplicity of infection of > or = 10. Indirect immunofluorescence assays using normal and confocal microscopy with flavivirus-reactive antibodies and WNV-infected neurons demonstrated viral antigen mostly associated with the plasma membrane and in the neurite processes. Treatment of WNV-infected C6/36, PC-12, or DRG cells with brefeldin A (BFA; a trans-Golgi inhibitor) or nocadazole (a beta-tubulin inhibitor) had little effect on viral maturation and secretion. Treatment of WNV-infected Vero cells with BFA resulted in a 1000-fold decrease in viral titer, but nocodazole had no effect. Our studies suggest that even though PC-12 and DRG neurons are mammalian cells, viral protein processing and maturation in these cells more closely resembles replication in C6/36 insect cells than in mammalian Vero cells. PMID- 15804956 TI - Human endogenous retrovirus (HERV)-W ENV and GAG proteins: physiological expression in human brain and pathophysiological modulation in multiple sclerosis lesions. AB - Antigen expression of a human endogenous retrovirus family, HERV-W, in normal human brain and multiple sclerosis lesions was studied by immunohistochemistry by three independent groups. The HERV-W multicopy family was identified in human DNA from the previously characterized multiple sclerosis-associated retroviral element (MSRV). A panel of antibodies against envelope (ENV) and capsid (GAG) antigens was tested. A physiological expression of GAG proteins in neuronal cells was observed in normal brain, whereas there was a striking accumulation of GAG antigen in axonal structures in demyelinated white matter from patients with MS. Prominent HERV-W GAG expression was also detected in endothelial cells of MS lesions from acute or actively demyelinating cases, a pattern not found in any control. A physiological expression of ENV proteins was detected in microglia in normal brain; however,a specific expression in macrophages was apparently restricted to early MS lesions. Thus, converging results from three groups confirm that GAG and ENV proteins encoded by the HERV-W multicopy gene family are expressed in cells of the central nervous system under normal conditions. Similar to HERV-W7q ENV (Syncitin), which is expressed in placenta and has been shown to have a physiological function in syncytio-trophoblast fusion, HERV-W GAG may thus also have a physiological function in human brain. This expression differs in MS lesions, which may either reflect differential regulation of inherited HERV-W copies, or expression of "infectious" MSRV copies. This is compatible with a pathophysiological role in MS, but also illustrates the ambivalence of such HERV antigens, which can be expressed in cell-specific patterns, under physiological or pathological conditions. PMID- 15804957 TI - Cognitive and motor deficits associated with HIV-2(287) infection in infant pigtailed macaques: a nonhuman primate model of pediatric neuro-AIDS. AB - Lentivirus-infected nonhuman primates exhibit behavioral and neurological pathology similar to human immunodeficiency virus (HIV)-infected humans and offer a means to examine the effects of lentivirus infection while controlling for confounding factors inherent in human populations. The purpose of this study was to examine cognitive and motor development in infant macaques vertically infected with HIV-2287. Subjects were 20 infant pigtail macaques (Macaca nemestrina); 8 controls born to uninfected dams, and 12 infants whose dams had been inoculated and infected with HIV-2287 in the third trimester of pregnancy. Eight of these pregnancies had undergone surgical procedures in the form of maternal amniotic catheters or maternal amniotic and fetal carotid artery and jugular vein catheters. Data indicated that catheterization had little or no impact on behavioral development. Seven infants were vertically infected (as measured by polymerase chain reaction (PCR) at birth) and five were not infected (as measured by PCR and coculture on repeated testing). Infected infants attained cognitive and motor milestones at significantly later ages than controls. Uninfected infants, born to infected dams, attained developmental milestones at later ages than controls on all tasks, but this reached statistical significance only for the Fine Motor Task. Attainment of milestones was not correlated with viral dose, maternal CD4+ levels at parturition or infant viral RNA levels at birth. Attainment of milestones was negatively correlated with infants' proportions of CD4+ lymphocytes at birth and significantly correlated with proportions of CD4+ lymphocytes 2 weeks after birth, indicating poorer performance in those infants with a more rapid CD4+ depletion. These cognitive and motor deficits closely resemble those observed in human infants and children infected with HIV and indicate that HIV-2287-infected infant macaques represent an excellent model of pediatric neuro-acquired immunodeficiency syndrome (neuroAIDS). PMID- 15804958 TI - Atypical radiological presentation of progressive multifocal leukoencephalopathy following liver transplantation. AB - Progressive multifocal leukoencephalopathy (PML), a demyelinating disease of the brain caused by JC virus (JCV), occurs following transplantation and other conditions associated with immunosuppression. On magnetic resonance imaging (MRI), PML lesions typically appear as hyperintense signal on T2-weighted and FLAIR images located in the subcortical white matter, which are devoid of contrast enhancement or mass effect. The prognosis is poor, but unusual inflammatory forms of PML characterized by contrast enhancement have been associated with a cellular immune response against JCV and a better prognosis. The authors report an atypical presentation of PML with contrast-enhancing lesions and mass effect on the MRI in a liver transplant recipient,who had a progressive course and fatal outcome. PMID- 15804959 TI - A case of a progressive multifocal leukoencephalopathy patient with four different JC virus transcriptional control region rearrangements in cerebrospinal fluid, blood, serum, and urine. AB - JC virus (JCV) is the etiological agent of progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease of the central nervous system (CNS). During the acquired immunodeficiency syndrome (AIDS) epidemic, it was the cause of the death in up to 8% of AIDS patients. The genomic organization of JCV and, in particular, the hypervariability of the transcriptional control region (TCR), a regulatory noncoding region, are well known. Given that the TCR plays a central role in the viral replication of JCV, a crucial role in the determination of the neurotropism and in the pathogenic capabilities of the virus is also suspected. Here the authors describe a case of PML that did not respond to highly active antiretroviral therapy (HAART) therapy. There was a simultaneous presence of JCV strains with four different TCR structures in urine, peripheral blood cells, serum, and cerebrospinal fluid (CSF) samples. These data confirmed that the presence of the archetype TCt is restricted to urine, while also suggesting that the degree of the rearrangement varies and increases from the peripheral blood to CSF. PMID- 15804960 TI - Genotypes of JC virus, DNA of cytomegalovirus, and proviral DNA of human immunodeficiency virus in eyes of acquired immunodeficiency syndrome patients. AB - JC virus (JCV) is a human polyomavirus that exists in at least eight different genotypes as a result of coevolution with different human populations all over the world. Well adapted to its host, it usually persists in the kidneys and possibly the brain. If the host becomes immunodeficient, JCV can cause the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML). There is increasing evidence that JCV is transactivated by cytomegalovirus (CMV) and the human immunodeficiency virus (HIV). Both CMV and HIV can infect the retina of acquired immunodeficiency syndrome (AIDS) patients, causing severe necrosis in the case of CMV retinitis or a mild HIV-associated vasculopathy, with bleeding and cotton wool spots. The authors therefore investigated by polymerase chain reaction (PCR) whether DNA of these three viruses was detectable in paraffin embedded eyes of AIDS patients with a clinical history of CMV retinitis. From a total of 65 eyes, JCV was detected in 21 (32%). Thirty-six (55%) were positive for CMV and 6 (9%) for proviral DNA of HIV. JCV and CMV were found in 13 eyes, JCV and HIV in 3 eyes, CMV and HIV in 1 eye, and DNA from all three viruses in 1 eye. The JCV genotypes were types 1A, 2A, 2E, 3, and 4. In 21 eyes of patients without AIDS, only one sample was JCV positive. In conclusion, JCV DNA can be detected in ocular tissue of AIDS patients at a significantly higher level than in eyes of nonimmunosuppressed patients. Further investigations will help to decide if JCV contributes to the retinopathy caused by CMV and HIV. PMID- 15804961 TI - Detection of interleukin 10 in cerebrospinal fluid of patients with subacute sclerosing panencephalitis. AB - Subacute sclerosing panencephalitis (SSPE) is a neurodegenerative disorder due to persistent measles virus infection, with high level of measles-specific antibodies in cerebrospinal fluid (CSF). To analyze whether such response arises from a TH2-biased response, the authors determined TH1 (interferon [IFN]-gamma) and TH2 (interleukin [IL]-4 and IL-10) cytokines in CSF, taken at diagnosis, of eight SSPE patients (median age, 57.5 month, range 42 to 76 months). All patients presented IL-10 (median 29.3 pg/ml, range 4.3 to 162 pg/ml), but not IL-4 (<10 pg/ml); only one case showed IFN-gamma (162 pg/ml). These results are consistent with a TH2 bias or with a local, anti-inflammatory or neuroprotective mechanism involving IL-10. PMID- 15804962 TI - Quantitative assessment of spasticity in human T-cell lymphotropic virus type I associated myelopathy/tropical spastic paraparesis. AB - People with human T-cell lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) develop spasticity. The authors examined 34 patients with HAM/TSP in Peru using a device that measures tone in the gastroc soleus-Achilles tendon unit and provides a quantitative spasticity assessment (QSA). Tone in the 34 patients was more than double that of women with asymptomatic HTLV-I infection. The device may help to track progression in HTLV-I infection. PMID- 15804964 TI - The application of reverse genetics technology in the study of rabies virus (RV) pathogenesis and for the development of novel RV vaccines. AB - Rabies is a central nervous system (CNS) disease that is almost invariably fatal. Neurotropism, neuroinvasiveness, and transsynaptic spread are the main features that determine the pathogenesis of rabies. Recent advances in rabies virus (RV) research, which made direct genetic manipulations of the RV genome possible, greatly improved the understanding of the role of different viral and host cell factors in the pathogenesis of rabies. Here the authors discuss molecular mechanisms associated with rabies RV infection and its spread to the CNS. PMID- 15804966 TI - The role of immune responses in the pathogenesis of rabies. AB - In the absence of treatment, infection with a variety of rabies virus strains most often results in a lethal outcome. This can be averted by prompt immunization following exposure demonstrating that the development of anti-rabies viral immunity prior to extensive infection of neurons is protective. Otherwise it might be expected that immune clearance of the virus would result in neurological sequelae. Thus, the capacity of a rabies virus to induce a protective immune response is a major, negative determinant of its pathogenicity and highly pathogenic rabies viruses have characteristics that avoid triggering protective immune responses. On the other hand, there is evidence that certain aspects of immunity may contribute to the pathogenesis of rabies under certain circumstances. The relationship between rabies virus and the immune system of the host is the focus of this review. PMID- 15804965 TI - Rabies virus receptors. AB - There is convincing in vitro evidence that the muscular form of the nicotinic acetylcholine receptor (nAChR), the neuronal cell adhesion molecule (NCAM), and the p75 neurotrophin receptor (p75NTR) bind rabies virus and/or facilitate rabies virus entry into cells. Other components of the cell membrane, such as gangliosides, may also participate in the entry of rabies virus. However, little is known of the role of these molecules in vivo. This review proposes a speculative model that accounts for the role of these different molecules in entry and trafficking of rabies virus into the nervous system. PMID- 15804967 TI - Pathophysiology of human paralytic rabies. AB - Furious rabies is a well-recognized clinical disorder in humans but the paralytic form is not as easily identified. The mechanisms responsible for the weakness and longer survival periods are not clear. Several hypotheses have been proposed, including rabies virus variants associated with a particular vector, location of wounds, incubation period, influence of prior rabies vaccination, and virus localization in the central nervous system (CNS). However, none of these have been substantiated. Regarding molecular analyses of rabies viruses isolated from both furious and paralytic rabies patients, only minor genetic variations with no specific patterns in glyco- (G), phospho- (P), and nucleoprotein (N) sequences have been identified and arginine 333 in G protein was present in all samples. Regional distribution of rabies virus antigenin rabies patients whose survival periods were 7 days or less and magnetic resonance imaging (MRI) of the CNS indicated brainstem and spinal cord as predilection sites regardless of clinical presentations. There are clinical, electrophysiological, and pathological indications that peripheral nerve dysfunction is responsible for weakness in paralytic rabies whereas in furious rabies, even in the absence of clinical weakness, abundant denervation potentials with normal sensory nerve conduction studies and proximal motor latencies suggest anterior horn cell dysfunction. The lack of cellular immunity to rabies virus antigen accompanied by an absence of cerebrospinal fluid (CSF) rabies neutralizing antibody in most paralytic rabies patients may argue against role of an immune response against rabies virus positive axons. Aberrant immune responses to peripheral nerve antigen, in particular those mediated by one or more cellular-dependent mechanisms, may be involved as is supported by the absence of putative anti-ganglioside antibodies commonly found in immune-mediated peripheral nerve diseases. Longer survival period in paralytic rabies may possibly be related to currently unidentified mechanism(s) on neuronal gene expression, required for virus transcription/replication and for maintaining neuronal survival. PMID- 15804968 TI - Neuronal dysfunction and death in rabies virus infection. AB - Because morphologic changes in natural rabies are usually relatively mild, it is thought that the severe clinical disease with a fatal outcome must be due to neuronal dysfunction of rabies virus-infected neurons. The precise bases of this functional impairment are unknown, and current knowledge on electro-physiological alterations, effects on ion channels and neurotransmission, and neurotoxicity are reviewed. Rabies virus may induce neuronal death, possibly through apoptotic mechanisms. Neuronal apoptosis has been observed in vitro and also in vivo under particular experimental conditions. The relevance of neuronal apoptosis in these situations to natural rabies has not yet been fully elucidated. PMID- 15804969 TI - Special edition of Microcirculation commemorating the 50th anniversary of the Microcirculatory Society, Inc. AB - This special edition of Microcirculation brings together the proceedings from the Symposium held in the spring of 2004 in the Natcher Auditorium on the campus of the National Institutes of Health to commemorate the 50th Anniversary of the Microcirculatory Society, Inc. Twelve brief reviews are introduced with the goals being to present a collective sense of the history of research in microcirculation, provide insight into where established topics of inquiry stand today, and to define emerging topics of investigation for future research efforts. PMID- 15804970 TI - Microvascular rheology and hemodynamics. AB - The goal of elucidating the biophysical and physiological basis of pressure-flow relations in the microcirculation has been a recurring theme since the first observations of capillary blood flow in living tissues. At the birth of the Microcirculatory Society, seminal observations on the heterogeneous distribution of blood cells in the microvasculature and the rheological properties of blood in small bore tubes raised many questions on the viscous properties of blood flow in the microcirculation that captured the attention of the Society's membership. It is now recognized that blood viscosity in small bore tubes may fall dramatically as shear rates are increased, and increase (dramatically with elevations in hematocrit. These relationships are strongly affected by blood cell deformability and concentration, red cell aggregation, and white cell interactions with the red cells anti endothelium. Increasing strength of red cell aggregation may result in sequestration of clumps of red cells with either reductions or increases in microvascular hematocrit dependent upon network topography. During red cell aggregation, resistance to flow may thus decrease with hematocrit reduction or increase due to redistribution of red cells. Blood cell adhesion to the microvessel wall may initiate flow reductions, as, for example, in the case of red cell adhesion to the endothelium in sickle cell disease, or leukocyte adhesion in inflammation. The endothelial glycocalyx has been shown to result from a balance of the biosynthesis of new glycans, and the enzymatic or shear dependent alterations in its composition. Flow-dependent reductions in the endothelial surface layer may thus affect the resistance to flow and/or the adhesion of red cells and/or leukocytes to the endothelium. Thus, future studies aimed at the molecular rheology of the endothelial surface layer may provide new insights into determinants of the resistance to flow. PMID- 15804971 TI - Microvascular solute and water transport. AB - OBJECTIVE: This review evaluates (1) the regulation of water and solute transport across the endothelial barrier in terms of pore theory and the glycocalyx junction-break model of capillary permeability; and (2) the mechanisms regulating permeability based on experiments using cultured endothelial cells and intact microvessels. CONCLUSIONS: The current form of the glycocalyx-junction-break model of capillary permeability describes the selectivity of the capillary wall (pore size) in terms of the space between the fibers of a quasi-periodic matrix on the endothelial cell surface, and the area for exchange (pore number) in terms of the length and frequency of breaks in the tight junction strands. An independent test of this model in a range of mammalian microvascular beds is new experimental evidence that the colloid osmotic pressure of plasma proteins is developed across the glycocalyx, not across the whole microvessel wall. We are beginning to understand that endothelial cells may change their phenotype in response to physical and chemical stresses. Such changes in phenotype may explain changes in the regulation of endothelial barrier function in intact microvessels that have previously been exposed to injury and differences in the regulation of contractile mechanisms between endothelial cells in vivo and in vitro. PMID- 15804972 TI - Regulation of blood flow in the microcirculation. AB - The regulation of blood flow has rich history of investigation and is exemplified in exercising skeletal muscle by a concerted interaction between striated muscle fibers and their microvascular supply. This review considers blood flow control in light of the regulation of capillary perfusion by and among terminal arterioles, the distribution of blood flow in arteriolar networks according to metabolic and hemodynamic feedback from active muscle fibers, and the balance between peak muscle blood flow and arterial blood pressure governed by sympathetic nerve activity. As metabolic demand increases,the locus of regulating oxygen delivery to muscle fibers "ascends" from terminal arterioles, through intermediate distributing arterioles, and into the proximal arterioles and feed arteries, which govern total flow into a muscle. At multiple levels, venules are positioned to provide feedback to nearby arterioles regarding the metabolic state of the tissue through the convection, production and diffusion of vasodilator stimuli. Electrical signals initiated on microvascular smooth muscle and endothelial cells can travel rapidly for millimeters through cell-to-cell conduction via gap junction channels, rapidly coordinating vasodilator responses that govern the distribution and magnitude of blood flow to active muscle fibers. Sympathetic constriction of proximal arterioles and feed arteries can restrict functional hyperemia while dilation prevails in distal arterioles to promote oxygen extraction. With vasomotor tone reflecting myogenic contraction of smooth muscle cells modulated by shear stress on the endothelium, the initiation of functional vasodilation and its modulation by sympathetic innervation dictate how and where blood flow is distributed in response to metabolic demand. A remarkable ensemble of signaling pathways underlies the integration of smooth muscle and endothelial cell function in microvascular networks. These pathways are being defined with refreshing new insight as novel approaches are applied to understanding the cellular and molecular mechanisms of blood flow control. PMID- 15804973 TI - Angiogenesis and microvascular remodeling: a brief history and future roadmap. AB - Angiogenesis and vessel remodeling determine the integrative control of the architectural structure and functional behaviors of the microcirculation over the lifetime of an organism. Vascular remodeling is the basis of promising therapeutic strategies, including vascularization of ischemic organs. The history of angiogenesis research is long-more than 250 years-and the Microcirculatory Society has been the birthplace of numerous techniques, assays, and scientific concepts that have stimulated massive research endeavors in the pharmaceutical and medical arena. At present, angiogenesis isa dynamic field in which the molecular genetic and proteomic components of the process are still being identified, while integrative systems approaches are once again being recognized as essential to understand microvascular assembly in vivo across multiple scales from cells to whole vessel networks. A short history of people and ideas in this field is presented, followed by discussion of emerging directions receiving intense attention today and major questions that remain unanswered. The primary conclusion is that the need for scientists trained in the integrative approaches nurtured by the Microcirculatory Society over the past 50 years has never been greater, as it is clear that a complete mechanistic understanding of vessel adaptation (based on genomic and proteomic supporting casts) will now require deeper studies of angiogenesis and microvascular remodeling in the exquisite complexity of the native microenvironment-the microcirculation. PMID- 15804974 TI - Oxygen transport and exchange in the microcirculation. AB - The cardiovascular system is responsible for maintaining an adequate convective delivery of oxygen to the smallest branches of the network of blood vessels-the microcirculation-from which oxygen passes to the parenchymal cells by passive diffusion. The aim of this brief review is to trace the development of the study of oxygen transport from the point of view of the microcirculation. August Krogh performed measurements that allowed him to use his keen insight to draw conclusions about oxygen transport that remained the foundations of this field for decades. After an extended period of neglect, Duling rekindled interest in the field of oxygen transport by discovering that substantial amounts of oxygen diffused from the arteriolar network. Subsequent investigations confirmed this finding ill various vascular beds and extended these studies to capillaries and venules. The important contributions of computational modeling and new techniques in intravital microscopy continue to lead to more advances in our understanding of the role of the microcirculation in the supply of oxygen to tissues. Current work is applying the concepts and principles learned in normal tissues to pathophysiological situations, as well as increasing our understanding of artificial oxygen carriers, oxygen sensing, and the connections between nitric oxide and oxygen transport. PMID- 15804975 TI - A new hypothesis for microvascular inflammation in shock and multiorgan failure: self-digestion by pancreatic enzymes. AB - Shock is accompanied by a severe inflammatory cascade in the microcirculation, the origin of which has been hypothesized in the past to be associated with specific mediators such as endotoxin, oxygen free radicals, nitric oxide, cytokines, and lipid products. But no intervention with clinical effectiveness has been derived from these ideas to date. The authors propose here a new hypothesis suggesting that degradative enzymes, synthesized in the pancreas as part of normal digestion, may play a central role in shock and multiorgan failure. These powerful enzymes have the ability to digest almost every biological material. Self-digestion (i.e. autodegradation) is prevented by compartmentalizing the fully activated degradative enzymes in the intestinal lumen by the mucosal barrier. In shock, maintenance of the mucosal barrier is impaired and it becomes permeable to pancreatic enzymes. Digestive enzymes thereby gain access to the wall of the intestine and initiate self-digestion of submucosal extracellular matrix proteins and interstitial cells. The process leads to generation and release of a host of strong inflammatory mediators. The authors hypothesize that inhibition of pancreatic enzymes in the lumen of tile intestine can serve to attenuate formation of these inflammatory mediators in ischemic tissues following hemorrhagic shock, and consequently prevent cell and tissue injury as well as multiorgan failure. PMID- 15804976 TI - Emerging topics in the regulation of leukocyte transendothelial migration. AB - This year marks the 50th Anniversary of the founding of the Microcirculatory Society. Since the formation of this society this field has witnessed tremendous progress in understanding the process of leukocyte recruitment during inflammation, injury, and immune reactions. This topic has been an important focus of many of the members of the Microcirculatory Society as well as our colleagues worldwide. The goal of this brief review is to bring attention to a few emerging topics in inflammation research. Here the focus is on one particular model of how one leukocyte type (PMN) can regulate the recruitment of a second different leukocyte type (T cell) and provide an outline of other aspects that bear on spatial and temporal behavior of specific leukocyte and endothelial cell adhesion molecules during leukocyte transmigration under dynamic shear flow in vitro. PMID- 15804977 TI - Therapeutic intervention in inflammatory diseases: a time and place for anti adhesion therapy. AB - The recruitment of leukocytes from the blood into tissue is central to the development and maintenance of the majority of inflammatory diseases. This multistep process requires a series of leukocyte-endothelial adhesive interactions, involving several families of adhesion molecules. Molecules that block these interactions have been targeted as potential therapeutic treatments for acute and chronic inflammatory diseases. However, many of the anti-adhesion therapy clinical trials have yielded disappointing outcomes. This review discusses some of the animal models that raise questions about the suitability of anti-adhesion therapy to treat certain inflammatory diseases. The authors suggest that it is crucial to understand the underlying mechanisms and time lines of leukocyte recruitment in each affected tissue and inflammatory disease to develop more effective anti-adhesion therapy. PMID- 15804978 TI - Integrins and regulation of the microcirculation: from arterioles to molecular studies using atomic force microscopy. AB - Integrins are an important class of receptors for extracellular matrix proteins that can mediate both force transmission, by virtue of their connections with the cell matrix and cytoskeleton; and signal transduction, resulting from the assemblages of signaling proteins that associate with focal contacts. Consequently, integrins have been proposed to be the mechanosensor in vascular smooth muscle and endothelial cells and to play a central role in mechanotransduction. In this regard, mechanical force is an important stimulus for many vascular functions, including contractile and relaxation processes,proliferation, migration, attachment, and cell phenotype determination. Collectively, these functions define physiological properties of the vasculature such as control of blood flow, capillary pressure,permeability, and peripheral vascular resistance, and play a role in pathophysiological processes like hypertension, diabetes, and arteriosclerosis. Our knowledge concerning how integrins sense and transduce physical forces into cellular signals and which integrins are involved is incomplete. Compared to other cell surface receptors, integrins have a relatively low affinity for their binding sites on the extracellular matrix and their affinity can be regulated. These characteristics of integrin-ligand interaction may facilitate dynamic processes such as cell migration, cell remodeling, and contractile activation in response to external forces. Important questions remain concerning the nature and origin of integrin mediated signaling in the vascular wall. PMID- 15804980 TI - Lymphatic biology and the microcirculation: past, present and future. AB - Because of the role that lymphatics have in fluid and macromolecular exchange, lymphatic function has been tightly tied to the study of the microcirculation for decades. Despite this, our understanding of many basic tenets of lymphatic function is far behind that of the blood vascular system. This is in part due to the difficulty inherent in working in small, thin-walled, clear lymphatic vessels and the relative lack of lymphatic specific molecular/cellular markers. The application of cellular and molecular tools to the field of lymphatic biology has recently produced some significant developments in lymphatic endothelial cell biology. These have propelled our understanding of lymphangiogenesis and related fields forward. Whereas the use of some of these techniques in lymphatic muscle biology has somewhat lagged behind those in the endothelium, recent developments in lymphatic muscle contractile and electrical physiology have also led to advances in our understanding of lymphatic transport function, particularly in the regulation of the intrinsic lymph pump. However, much work remains to be done. This paper reviews significant developments in lymphatic biology and discusses areas where further development of lymphatic biology via classical, cellular, and molecular approaches is needed to significantly advance our understanding of lymphatic physiology. PMID- 15804981 TI - Promoting early return to pre-injury job using a rebar-tying machine. PMID- 15804979 TI - Potassium channels in the peripheral microcirculation. AB - Vascular smooth muscle (VSM) cells, endothelial cells (EC), and pericytes that form the walls of vessels in the microcirculation express a diverse array of ion channels that play an important role in the function of these cells and the microcirculation in both health and disease. This brief review focuses on the K+ channels expressed in smooth muscle and endothelial cells in arterioles. Microvascular VSM cells express at least four different classes of K+ channels, including inward-rectifier K+ channels (Kin), ATP-sensitive K+ channels (KATP), voltage-gated K+ channels (Kv), and large conductance Ca2+-activated K+ channels (BKCa). VSM KIR participate in dilation induced by elevated extracellular K+ and may also be activated by C-type natriuretic peptide, a putative endothelium derived hyperpolarizing factor (EDHF). Vasodilators acting through cAMP or cGMP signaling pathways in VSM may open KATP, Kv, and BKCa, causing membrane hyperpolarization and vasodilation. VSMBKc. may also be activated by epoxides of arachidonic acid (EETs) identified as EDHF in some systems. Conversely, vasoconstrictors may close KATP, Kv, and BKCa through protein kinase C, Rho kinase, or c-Src pathways and contribute to VSM depolarization and vasoconstriction. At the same time Kv and BKCa act in a negative feedback manner to limit depolarization and prevent vasospasm. Microvascular EC express at least 5 classes of K+ channels, including small (sKCa) and intermediate(IKCa) conductance Ca2+-activated K+ channels, Kin, KATP, and Kv. Both sK and IK are opened by endothelium-dependent vasodilators that increase EC intracellular Ca2+ to cause membrane hyper-polarization that may be conducted through myoendothelial gap junctions to hyperpolarize and relax arteriolar VSM. KIR may serve to amplify sKCa- and IKCa-induced hyperpolarization and allow active transmission of hyperpolarization along EC through gap junctions. EC KIR channels may also be opened by elevated extracellular K+ and participate in K+-induced vasodilation. EC KATP channels may be activated by vasodilators as in VSM. Kv channels may provide a negative feedback mechanism to limit depolarization in some endothelial cells. PMID- 15804982 TI - WBGT clothing adjustments for four clothing ensembles under three relative humidity levels. AB - Threshold limit values for heat stress and strain are based on an upper limit wet bulb globe temperature (WBGT) for ordinary work clothes, with clothing adjustment factors (CAF) for other clothing ensembles. The purpose of this study was to determine the CAF for four clothing ensembles (Cotton Coveralls, Tyvek 1424 Coveralls, NexGen Coveralls, and Tychem QC Coveralls) against a baseline of cotton work clothes and to determine what effect relative humidity may have. A climatic chamber was used to slowly increase the level of heat stress by increasing air temperature at three levels of relative humidity (20%, 50%, and 70%). Study participants wore one of the five ensembles while walking on a treadmill at a moderate metabolic rate of 155 W m-2 (about 300 W). Physiological data and environmental data were collected. When the participant's core temperature reached a steady state, the dry bulb temperature was increased at constant relative humidity. The point at which the core temperature began to increase was defined as the inflection point. The environmental temperature recorded 5 min before the inflection point was used to calculate the critical WBGT for each ensemble. A three-way analysis of variance with ensemble by humidity protocol interactions and a multiple comparison test were used to make comparisons among the mean values. Only the vapor-barrier ensemble (Tychem QC) demonstrated an interaction with humidity level. The following CAFs are proposed: Cotton Coveralls (0 degrees C-WBGT), Tyvek 1424 Coveralls (+1), NexGen Coveralls (+2), and Tychem QC Coveralls (+10). PMID- 15804994 TI - Secondary neoplasms after radiotherapy for a childhood solid tumor. AB - This study was conducted to determine the outcome of patients who develop a second neoplasm after radiotherapy (RT) for a childhood solid tumor. From 1956 to 1998, 429 children with a malignant solid tumor were treated at a single radiation oncology facility. The medical records and radiotherapy charts were reviewed to determine if the patient developed a secondary neoplasm after treatment for malignancy. Twenty-three (5.4%) patients developed a secondary neoplasm. There were 12 males and 11 females with a median age at RT of 6.6 years (range, 2 months to 20 years). There were 14 malignant neoplasms in 13 (3.0%) and 14 benign neoplasms in 11 patients (2.6%). The types of initial solid tumors treated with RT were Ewing sarcoma in 6, Wilms tumor in 6, medulloblastoma in 5, neuroblastoma in 3, and other in 3. Median RT dose was 45 Gy (range, 12.3 to 60 Gy) using 4 MV in 9, 1.25 MV in 8, 250 KV in 4, and 6 MV photons in 1 patient. One child was treated using 15-MeV electrons. Fourteen had chemotherapy. Median follow-up was 23.2 years (range, 5.3 to 44.4 years). For the 14 malignant neoplasms, the median time interval from initial tumor to second malignancy was 10.1 years. The 14 second malignant neoplasms (SMN) were osteosarcoma in 3, breast carcinoma in 2, melanoma in 2, malignant fibrous histiocytoma in 1, dermatofibrosarcoma in 1, leiomyosarcoma in 1, mucoepidermoid carcinoma in 1, colon cancer in 1, chronic myelogenous leukemia in 1, and basal cell carcinoma in 1. Ten of the 14 SMN (71%) were at the edge or inside the RT field. The 5- and 10 year overall survival rate after diagnosis of an SMN was 69.2%; it was 70% for children with a SMN at the edge or inside the RT field and 66.7% for those outside of the RT field. The 14 benign neoplasms appeared at a median time of 16.9 years and included cervical intraepithelial neoplasia in 3, osteochondroma in 3, thyroid adenoma in 1, duodenal adenoma in 1, lipoma in 1, cherry angioma in 1, uterine leiomyoma in 1, ovarian cystadenofibroma in 1, and giant cell tumor in 1. Only 5 (36%) of the 14 benign tumors occurred in the RT field, with osteochondroma being the most common. Of 189 deaths occurring in 429 patients, only 3 (1.6%) were secondary to radiation-induced malignancy. Not all SMN in children receiving RT occur in the irradiated field. More than two-thirds of children with a radiation-induced malignancy are alive 10 years after the diagnosis of a SMN. PMID- 15804995 TI - A prospective randomized trial of the antiemetic efficacy and cost-effectiveness of intravenous and orally disintegrating tablet of ondansetron in children with cancer. AB - Orally disintegrating tablet (ODT) of ondansetron is a new formulation, which instantaneously disintegrates and disperses in the saliva without need for ingestion of a liquid. This makes the formulation suitable for administration in children. The objective of this study was to compare the relative efficacy and cost of ODT and intravenous (IV) formulation of ondansetron in controlling nausea and vomiting in children receiving chemotherapy regimens without cisplatin. This prospective randomized trial was performed in a single institution to compare ODT and IV formulation of ondansetron for the prevention of acute emesis in a group of 22 children. Study agents were administered 30 min before chemotherapy and 12 hourly after chemotherapy (5 mg/m2 IV or 4-8 mg oral according to body surface area in 56 and 39 courses, respectively). After randomization, IV formulation was administered to some children instead of ODT due to unavailability of this formulation. Complete and major control of emesis was obtained in 92% of patients in the IV group and 93% of patients in the ODT group. In 56 courses with grade III-IV emetogenicity, complete response rates were not different between the two treatment arms. In the courses without corticosteroids complete response rates were not also different between the two arms. The mean costs per successfully controlled courses were 121.3 USD for the IV formulation whereas 63.2 USD for the ODT formulation. The results of this study confirmed that ODT formulation of ondansetron is a safe, well-tolerated, and cost-effective antiemetic for children during non-cisplatin-containing moderately and highly emetogenic chemotherapy. PMID- 15804996 TI - The geldanamycin derivative 17-AAG decreases VEGF secretion and leukemia growth support by trisomy 8 myelodysplastic syndrome bone marrow stromal cells. AB - Stromal cells from a child with constitutional trisomy 8 who developed myelodysplastic syndrome (MDS) were found to produce abnormal levels of various cytokines, including VEGF, and supported the growth of leukemic cells in co culture assays. This study shows that the geldanamycin derivative 17-AAG effectively reduces the VEGF expression by MDS stromal cells. In co-culture experiments this agent also blocks the ability of the MDS stromal cells to stimulate the growth of leukemic cells. These data provide important initial evidence for the effect of 17-AAG on the marrow microenvironment and its potential use in the treatment of MDS and leukemia. PMID- 15804997 TI - Craving for ice and iron-deficiency anemia: a case series from Oman. AB - Pagophagia, or the practice of consuming ice, is a particular expression of the more general phenomenon of pica. Pagophagia is a complex behavioral phenomenon arising from the interplay of biochemical, hematological, psychological, and cultural factors. This compulsive dietary aberration is observed in many children and pregnant women worldwide. The authors report 3 cases of severe iron deficiency anemia with a serum ferritin level of 2-3 ng/mL, in which the patients were consuming 2 trays and many bags of ice per day. Following treatment with iron therapy, pagophagia spontaneously resolved within 2 weeks. It is a commonly missed problem. Pediatricians should be alert to this phenomena and its association with iron-deficiency anemia. PMID- 15804998 TI - Interferon therapy in congenital dyserythropoietic anemia type I/II. AB - Congenital dyserythropoietic anemia (CDA) is a rare disorder, characterized by the association of ineffective erythropoiesis, variable degree of anemia, and erythroblastic morphological abnormalities. alpha-Interferon has been reported to be effective in type I CDA, but efficacy in other types of CDA is uncertain. Encouraged by the reports, we evaluated the efficacy of alpha-interferon in 6 children with CDA. Diagnosis of CDA was established on the basis of clinical profile, distinct morphological findings on light microscopy, and the Ham's test, following the exclusion of the more common causes of hemolysis. Erythrocyte agglutinability and lysis to anti-i and anti-l sera, electron microscopy, and SDS polyacrylamide gel electrophoresis were not performed, due to nonavailability. There were 3 cases, each, with type I and type II CDA. The mean age was 5.5 years (range: 6 months to 11.5 years). Five of the 6 patients were transfusion dependent. alpha-Interferon was administered subcutaneously for a mean duration of 19 weeks (range: 12-30). The dose ranged from 2.6 to 6.5 million IU/m2/dose. The frequency of injections varied from thrice weekly to alternate days. No favorable effect on hemoglobin, reticulocyte count, or transfusion frequency was observed. alpha-Interferon therapy was found to be ineffective in all the patients. These observations question the use of interferon in CDA until further studies in a larger number of patients establish its efficacy. PMID- 15804999 TI - Platelet aggregation in term and preterm newborns. AB - The platelets of newborns have a hyporeactive period. This period, during which the platelet count is normal but their functions are deficient, is called transient platelet hyporeactivity of newborns. The platelet functions and their normalizing process of term and preterm neonates are investigated. Twenty term and 20 preterm (gestational age <37 weeks) newborns were enrolled in the study. Twenty-eight healthy children aged 2 months to 3 years old participated in the study as the control group. Healthy newborns were followed for 15 days after birth longitudinally in 3 periods: period 1 (0-4 days), period 2 (5-9 days), period 3 (10-15 days). Aggregation studies were performed from whole blood samples. Whole blood aggregation was measured by the impedance method. Transient hyporeactivity of platelets was found in term and preterm groups, and there was no difference between term and preterms. Platelets of newborns gained their normal functions at postnatal 10-14 days. The results show that hyporeactivity of platelets during the first 9 days of life is physiological and transient. PMID- 15805000 TI - Amantadine in chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is a rare genetically determined immunodeficiency. Neutrophils from CGD patients show a defective killing of phagocytosed fungi and bacteria, due not only to an impairment in oxidative burst, but also to absence of normal pH value within phagocytic vacuole following phagocytosis. Because a weak base such as amantadine could potentially reverse these pH abnormalities, the authors used this drug to treat 2 CGD patients. They observed modifications of both phagosomal pH and killing activity on their neutrophils compared to those of healthy controls. Since the drug has been employed, the patients have not developed new infections, suggesting a role of amantadine as a part of CGD prophylactic regimen. These results suggest the opportunity of testing the drug in larger studies. PMID- 15805001 TI - Quantitative thallium-201 scintigraphy in childhood osteosarcoma: Comparison with technetuim-99m MDP and magnetic resonance imaging in the evaluation of chemotherapeutic response. AB - The aim of this study was to assess early (15 min) and late 2 h) thallium-201 (201Tl) uptake in children with osteosarcoma and to compare these findings with magnetic resonance imaging (MRI) and technetium-99m methylenediphosphonate (99mTc MDP), with emphasis on evaluating tumor viability before and after chemotherapy. Fifteen patients with biopsy-proven osteosarcoma received standard preoperative chemotherapy with a combination of cisplatin, Adriamycin, and high-dose methotrexate. Their ages ranged between 7 and 18 years (median 14.5 years). All patients had 201Tl, 99mTc MDP, and MRI studies. Thallium scintigraphy was performed at 15 min and 2 h after IV injection of 92 MBq of thallium. Thallium uptake ratio was calculated by dividing the count density of the lesion (L) by that of the controlateral normal (N) area. The percent reduction of 201Tl uptake ratio (alteration ratio) was calculated by [100x(prechemotherapy L/N postchemotherapy L/N)/prechemotherapy L/N]. Pathologic changes were graded on the basis of % tumor necrosis as defined histologically. Scintigraphic comparisons demonstrated a high-degree of correlation with late 201Tl alteration ration and poor correlation with both early 201Tl and 99mTc MDP alteration ratios. Late 201Tl images were superior to early 201Tl, 99mTc MDP, and MRI in predicting tumor response to chemotherapy as determined by % tumor necrosis (p<.01). The authors found that late 201Tl was an accurate test for evaluating the response to specific therapeutic regimens and it can be useful planning surgery or choosing alternative chemotherapeutic regimens. PMID- 15805002 TI - Heterozygous beta-thalassemia and homozygous H63D hemochromatosis in a child: an 18-year follow-up. AB - At age of 3.2 years routine blood analysis showed the presence of a beta thalassemic trait with unexpected high level of serum iron and high transferrin saturation. Hematological follow-up confirmed the moderate degree of anemia and persisting high levels of iron indices throughout the years with a progressive increase of serum ferritin. At the age of 19 years the patient was diagnosed homozygous for HC63D HFE. The patient referred by us confirm the possibility of precocious alteration of iron indices in patients with heterozygosity for beta thalassemia inherited together with HFE mutations. This observation suggests that any children with thalassemic trait with increased transferrin saturation and/or serum ferritin might be investigated for the presence of the hemocromatosis genes in order to detect the disease before any clinical manifestation and even before organ iron loading. PMID- 15805003 TI - Acral erythema caused by high-dose methotrexate therapy in patients with osteogenic sarcoma. AB - High doses of methotrexate (MTX) have been incorporated in the treatment of osteogenic sarcoma since the 1970s. Unfortunately, high-dose MTX (HD-MTX) can cause untoward side effects that may complicate the proper management of these patients. Two cases of MTX-induced acral erythema are described and the possible implications of this complication are reviewed, with a discussion of the pathophysiology of this adverse effect. Two young female patients suffering from osteogenic sarcoma received HD-MTX (12 g/m2) according to the chemotherapeutic protocol. A 17-year-old patient with osteogenic sarcoma of the head developed painful acral erythema of her palms and soles after the fifth dose of MTX, and a 22-year-old patient with osteosarcoma of the femur developed severe acral erythema after the sixth dose. The severity of the reaction was significant enough to cancel further treatment with HD-MTX in both patients. All symptoms resolved without sequellae several weeks later in both cases. Acral erythema may pose a significant albeit transient phenomenon adversely influencing chemotherapy in patients suffering from osteosarcoma and treated with HD-MTX. The pathophysiology of this side effect is not completely understood and further investigation of the pharmacokinetics of HD-MTX in this patient population is needed. PMID- 15805004 TI - Synchronous multicentric giant cell tumor in a 16-year-old boy. AB - Synchronous multicentric giant cell tumor of the bone is a rare variant of a lesion appearing during childhood. The authors report clinical, radiological, and pathological features of a 16-year-old boy who was diagnosed with synchronous multicentric giant cell tumor, which originated in the right distal femur and the left fibula. PMID- 15805005 TI - SECONDARY LEIOMYOSARCOMAS: A Report of 4 Cases. AB - Survivors of childhood cancer risk developing second malignant neoplasms (SMN). a small proportion of SMNs is represented by soft tissue sarcoma (STS), including leiomyosarcoma. The files of the STS Italian Cooperative Group (ICG) were reviewed and 4 patients with secondary leiomyosarcoma were identified, only two of them still alive. In 3 cases, the leiomyosarcoma occurred in the radiation field. The authors found no clear differences between primary and secondary leiomyosarcomas, but leiomyosarcoma proved to be the most frequent SMN registered by the ICG, confirming the need for further studies to identify any distinctive features or syndromes. PMID- 15805007 TI - Exploring rate variation among and within sites in a densely sampled tree: species level phylogenetics of north american tiger beetles (genus cicindela). AB - Species-level phylogenetic studies require fast-evolving nucleotide positions to resolve relationships among close relatives, but these sites may be highly homoplastic and perhaps uninformative or even misleading deeper in the tree. Here we describe a species-level analysis of tiger beetles in the genus Cicindela (Coleoptera: Cicindelidae) for 132 terminal taxa and 1897 nucleotide positions from three regions of mtDNA, comprising 75% coverage of species occurring in North America. Evenly weighted parsimony analysis recovered four major clades representing radiations confined to North and Central America. Relationships near the tips were well supported but signal was contradictory at deeper nodes. Two major categories (3rd positions and all others) can be distinguished in likelihood analysis of character variation, of which only the fast-changing 3rd position characters were affected by saturation. However, their downweighting under a variety of criteria did not improve the tree topology at basal nodes. There was weak conflict between 3rd and non-3rd position characters deep in the tree, but support levels declined towards the root for all categories, even on trees that were reconstructed from 3rd and non-3rd positions separately. Statistical analysis of parsimony-based character transitions along branches showed a largely homogeneous distribution of change along the root-to-tip axis. The comparison of character transitions among the four major portions of the tree revealed deviations from stochastic distribution for the non-3rd positions, but not for 3rd positions. Hence, variability of functionally constrained non-3rd positions differs between clades and may be dependent on the character states at other sites, consistent with the covarion model of molecular evolution. The results suggest that some properties of 3rd positions are less problematic for phylogenetic reconstruction than other categories despite their high total homoplasy. In densely sampled data sets of closely related species, the disadvantages of weighting schemes according to homoplasy levels outweigh the benefits, showing the difficulty of devising meaningful weighting schemes that are applicable universally throughout the tree. PMID- 15805008 TI - Phylogeny and biogeography of exacum (gentianaceae): a disjunctive distribution in the Indian ocean basin resulted from long distance dispersal and extensive radiation. AB - Disjunctive distributions across paleotropical regions in the Indian Ocean Basin (IOB) often invoke dispersal/vicariance debates. Exacum (Gentianaceae, tribe Exaceae) species are spread around the IOB, in Africa, Madagascar, Socotra, the Arabian peninsula, Sri Lanka, India, the Himalayas, mainland Southeast Asia including southern China and Malaysia, and northern Australia. The distribution of this genus was suggested to be a typical example of vicariance resulting from the breakup of the Gondwanan supercontinent. The molecular phylogeny of Exacum is in principle congruent with morphological conclusions and shows a pattern that resembles a vicariance scenario with rapid divergence among lineages, but our molecular dating analysis demonstrates that the radiation is too recent to be associated with the Gondwanan continental breakup. We used our dating analysis to test the results of DIVA and found that the program predicted impossible vicariance events. Ancestral area reconstruction suggests that Exacum originated in Madagascar, and divergence dating suggests its origin was not before the Eocene. The Madagascan progenitor, the most recent common ancestor of Exacum, colonized Sri Lanka and southern India via long-distance dispersals. This colonizer underwent an extensive range expansion and spread to Socotra-Arabia, northern India, and mainland Southeast Asia in the northern IOB when it was warm and humid in these regions. This widespread common ancestor retreated subsequently from most parts of these regions and survived in isolation in Socotra-Arabia, southern India-Sri Lanka, and perhaps mainland Southeast Asia, possibly as a consequence of drastic climatic changes, particularly the spreading drought during the Neogene. Secondary diversification from these surviving centers and Madagascar resulted in the extant main lineages of the genus. The vicariance-like pattern shown by the phylogeny appears to have resulted from long distance dispersals followed by extensive range expansion and subsequent fragmentation. The extant African species E. oldenlandioides is confirmed to be recently dispersed from Madagascar. PMID- 15805009 TI - Is homoplasy or lineage sorting the source of incongruent mtdna and nuclear gene trees in the stiff-tailed ducks (Nomonyx-Oxyura)? AB - We evaluated the potential effects of homoplasy, ancestral polymorphism, and hybridization as obstacles to resolving phylogenetic relationships within Nomonyx Oxyura stiff-tailed ducks (Oxyurinae; subtribe Oxyurina). Mitochondrial DNA (mtDNA) control region sequences from 94 individuals supported monophyly of mtDNA haplotypes for each of the six species and provided no evidence of extant incomplete lineage sorting or inter-specific hybridization. The ruddy ducks (O. j. jamaicensis,O. j. andina, O. j. ferruginea) are each others' closest relatives, but the lack of shared haplotypes between O. j. jamaicensis and O. j. ferruginea suggests long-standing historical isolation. In contrast, O. j. andina shares haplotypes with O. j. jamaicensis and O. j. ferruginea, which supports Todd's (1979) and Fjeldsa's (1986) hypothesis that O. j. andina is an intergrade or hybrid subspecies of O. j. jamaicensis and O. j. ferruginea. Control region data and a much larger data set composed of approximately 8800 base pairs of mitochondrial and nuclear sequence for each species indicate that the two New World species, O. vittata and O. jamaicensis, branch basally within Oxyura. A clade of three Old World species (O. australis, O. maccoa, O. leucocephala) is well supported, but different loci and also different characters within the mtDNA data support three different resolutions of the Old World clade, yielding an essentially unresolved trichotomy. Fundamentally different factors limited the resolution of the mtDNA and nuclear gene trees. Gene trees for most nuclear loci were unresolved due to slow rates of mutation and a lack of informative variation, whereas uncertain resolution of the mtDNA gene tree was due to homoplasy. Within the mtDNA, approximately equal numbers of characters supported each of three possible resolutions. Parametric and nonparametric bootstrap analyses suggest that resolution of the mtDNA tree based on ~4300 bp per taxon is uncertain but that complete mtDNA sequences would yield a fully resolved gene tree. A short internode separating O. leucocephala from (O. australis, O. maccoa) in the best mtDNA tree combined with long terminal branches and substantial rate variation among nucleotide sites allowed the small number of changes occurring on the internode to be obscured by homoplasy in a significant portion of simulated data sets. Although most nuclear loci were uninformative, two loci supported a resolution of the Old World clade (O. maccoa, O. leucocephala) that is incongruent with the best mtDNA tree. Thus, incongruence between nuclear and mtDNA trees may be due to random sorting of ancestral lineages during the short internode, homoplasy in the mtDNA data, or both. The Oxyura trichotomy represents a difficult though likely common problem in molecular systematics. Given a short internode, the mtDNA tree has a greater chance of being congruent with the history of speciation because its effective population size (N(e)) is one-quarter that of any nuclear locus, but its resolution is more likely to be obscured by homoplasy. In contrast, gene trees for more slowly evolving nuclear loci will be difficult to resolve due to a lack of substitutions during the internode, and when resolved are more likely to be incongruent with the species history due to the stochastic effects of lineage sorting. We suggest that researchers consider first whether independent gene trees are adequately resolved and then whether those trees are congruent with the species history. In the case of Oxyura, the answer to both questions may be no. Complete mtDNA sequences combined with data from a very large number of nuclear loci may be the only way to resolve such trichotomies. PMID- 15805010 TI - Biogeographic interpretation of splits graphs: least squares optimization of branch lengths. AB - Although most often used to represent phylogenetic uncertainty, network methods are also potentially useful for describing the phylogenetic complexity expected to characterize recent species radiations. One network method with particular advantages in this context is split decomposition. However, in its standard implementation this approach is limited by a conservative criterion for branch length estimation. Here we extend the utility of split decomposition by introducing a least squares optimization technique for correcting branch lengths that may be underestimated by the standard implementation. This optimization of branch lengths is generally expected to improve divergence time estimates calculated from splits graphs. We illustrate the effect of least squares optimization on such estimates using the Australasian Myosotis and the Hawaiian silversword alliance as examples. We also discuss the biogeographic interpretation and limitations of splits graphs. PMID- 15805011 TI - Visualizing conflicting evolutionary hypotheses in large collections of trees: using consensus networks to study the origins of placentals and hexapods. AB - Many phylogenetic methods produce large collections of trees as opposed to a single tree, which allows the exploration of support for various evolutionary hypotheses. However, to be useful, the information contained in large collections of trees should be summarized; frequently this is achieved by constructing a consensus tree. Consensus trees display only those signals that are present in a large proportion of the trees. However, by their very nature consensus trees require that any conflicts between the trees are necessarily disregarded. We present a method that extends the notion of consensus trees to allow the visualization of conflicting hypotheses in a consensus network. We demonstrate the utility of this method in highlighting differences amongst maximum likelihood bootstrap values and Bayesian posterior probabilities in the placental mammal phylogeny, and also in comparing the phylogenetic signal contained in amino acid versus nucleotide characters for hexapod monophyly. PMID- 15805012 TI - Mitochondrial phylogenetics and evolution of mysticete whales. AB - The phylogenetic relationships among baleen whales (Order: Cetacea) remain uncertain despite extensive research in cetacean molecular phylogenetics and a potential morphological sample size of over 2 million animals harvested. Questions remain regarding the number of species and the monophyly of genera, as well as higher order relationships. Here, we approach mysticete phylogeny with complete mitochondrial genome sequence analysis. We determined complete mtDNA sequences of 10 extant Mysticeti species, inferred their phylogenetic relationships, and estimated node divergence times. The mtDNA sequence analysis concurs with previous molecular studies in the ordering of the principal branches, with Balaenidae (right whales) as sister to all other mysticetes base, followed by Neobalaenidae (pygmy right whale), Eschrichtiidae (gray whale), and finally Balaenopteridae (rorquals + humpback whale). The mtDNA analysis further suggests that four lineages exist within the clade of Eschrichtiidae + Balaenopteridae, including a sister relationship between the humpback and fin whales, and a monophyletic group formed by the blue, sei, and Bryde's whales, each of which represents a newly recognized phylogenetic relationship in Mysticeti. We also estimated the divergence times of all extant mysticete species, accounting for evolutionary rate heterogeneity among lineages. When the mtDNA divergence estimates are compared with the mysticete fossil record, several lineages have molecular divergence estimates strikingly older than indicated by paleontological data. We suggest this discrepancy reflects both a large amount of ancestral polymorphism and long generation times of ancestral baleen whale populations. PMID- 15805013 TI - Ontogeny discombobulates phylogeny: paedomorphosis and higher-level salamander relationships. AB - Evolutionary developmental biology ("evo-devo") has revolutionized evolutionary biology but has had relatively little impact on systematics. We show that similar large-scale developmental changes in distantly related lineages can dramatically mislead phylogenetic analyses based on morphological data. Salamanders are important model systems in many fields of biology and are of special interest in that many species are paedomorphic and thus never complete metamorphosis. A recent study of higher-level salamander phylogeny placed most paedomorphic families in a single clade based on morphological data. Here, we use new molecular and morphological data to show that this result most likely was caused by the misleading effects of paedomorphosis. We also provide a well-supported estimate of higher-level salamander relationships based on combined molecular and morphological data. Many authors have suggested that paedomorphosis may be problematic in studies of salamander phylogeny, but this hypothesis has never been tested with a rigorous phylogenetic analysis. We find that the misleading effects of paedomorphosis on phylogenetic analysis go beyond the sharing of homoplastic larval traits by paedomorphic adults, and the problem therefore is not solved by simply excluding suspected paedomorphic characters. Instead, two additional factors are critically important in causing paedomorphic species to be phylogenetically "misplaced": (1) the absence of clade-specific synapomorphies that develop during metamorphosis in nonpaedomorphic taxa and allow their "correct" placement and (2) parallel adaptive changes associated with the aquatic habitat of the larval stage. Our results suggest that the effects of paedomorphosis on phylogenetic analyses may be complex, difficult to detect, and can lead to results that are both wrong and statistically well supported by parsimony and Bayesian analyses. PMID- 15805014 TI - Archaeobatrachian paraphyly and pangaean diversification of crown-group frogs. AB - Current models for the early diversification of living frogs inferred from morphological, ontogenetic, or DNA sequence data invoke very different scenarios of character evolution and biogeography. To explore central controversies on the phylogeny of Anura, we analyzed nearly 4000 base pairs of mitochondrial and nuclear DNA for the major frog lineages. Likelihood-based analyses of this data set are congruent with morphological evidence in supporting a paraphyletic arrangement of archaeobatrachian frogs, with an (Ascaphus + Leiopelma) clade as the sister-group of all other living anurans. The stability of this outcome is reinforced by screening for phylogenetic bias resulting from site-specific rate variation, homoplasy, or the obligatory use of distantly related outgroups. Twenty-one alternative branching and rooting hypotheses were evaluated using a nonparametric multicomparison test and parametric bootstrapping. Relaxed molecular clock estimates situate the emergence of crown-group anurans in the Triassic, approximately 55 million years prior to their first appearance in the fossil record. The existence of at least four extant frog lineages on the supercontinent Pangaea before its breakup gains support from the estimation that three early splits between Laurasia- and Gondwana-associated families coincide with the initial rifting of these landmasses. This observation outlines the potential significance of this breakup event in the formation of separate Mesozoic faunal assemblages in both hemispheres. PMID- 15805015 TI - The effect of paralogous lineages on the application of reconciliation analysis by cophylogeny mapping. AB - Paralogy defines similarity caused by duplication rather than common descent and is well known in the case of paralogous gene copies within a single genome. The term is here extended to paralogous lineages of associates within a single host. The phylogenies of four genera within the Herpesviridae were reconciled with host phylogenies using cophylogenetic mapping. The observed correspondence for each pair of phylogenies was evaluated through randomization of the viral phylogeny and demonstrated to be greater than expected by chance. A simulation study was then carried out to assess the influence of paralogous lineages on the efficacy of reconciliation analysis. Combining viral taxa from different genera that infected common hosts introduced incongruence into the cophylogenies and reduced both the minimum and maximum observed number of codivergence events relative to the initial analysis of orthologous clades. However, at an average sample size this did not alter the fundamental significance of observed correspondence. With smaller sample sizes, the number of orthologous taxa selected at random from the pool of taxa was reduced. False-negative results then increased in proportion from 0.02 to 0.33. These results demonstrated that reconciliation analysis is robust under conditions of paralogy at "normal" sample sizes but is adversely affected by a combination of paralogy and low sample size. Consideration of phylogenies for Papillomavirus, Atadenovirus, and Mastadenovirus suggest that paralogous lineages may be a widespread phenomenon among DNA viruses and that duplication irrespective of host speciation is an important cause of viral diversification. PMID- 15805016 TI - Missing the forest for the trees: phylogenetic compression and its implications for inferring complex evolutionary histories. AB - Phylogenetic tree reconstruction is difficult in the presence of lateral gene transfer and other processes generating conflicting signals. We develop a new approach to this problem using ideas borrowed from algorithmic information theory. It selects the hypothesis that simultaneously minimizes the descriptive complexity of the tree(s) plus the data when encoded using those tree(s). In practice this is the hypothesis that can compress the data the most. We show not only that phylogenetic compression is an efficient method for encoding most phylogenetic data sets and is more efficient than compression schemes designed for single sequences, but also that it provides a clear information theoretic rule for determining when a collection of conflicting trees is a better explanation of the data than a single tree. By casting the parsimony problem in this more general framework, we also conclude that the so-called total-evidence tree--the tree constructed from all the data simultaneously--is not always the most economical explanation of the data. PMID- 15805017 TI - Morphology's role in phylogeny reconstruction: perspectives from paleontology. PMID- 15805025 TI - The regulation of clinical photographers. AB - A long-held ambition for the regulation of clinical photographers is soon to be realized: in September 2004 the Health Professions Council formally approved an application by the Council for the Accreditation of Medical Illustration Practitioners, and it is anticipated that clinical photographers will be amongst the first group of healthcare scientists to join the national state register in 2005. This paper describes the application process and the ramifications for the profession. PMID- 15805026 TI - Implementation of standardized medical photography for cleft lip and palate audit. AB - Much has been published relating to the treatment and surgical outcome of cleft lip and palate disorders. Clinical audit is one of the most important tools for assessing the quality of care provided, with medical photography an invaluable component of this process. The Clinical Standards Advisory Group Report 1998 recommended that cleft lip and palate patients should be audited when 0 (under 1 year of age), and then at the ages of 5, 10, 15 and 20 years. For both audit and research purposes, medical photographs need to be accurate and of a consistently high standard. This paper describes the development of a standardized photographic protocol for cleft patients of audit age, to the benefit of both the multidisciplinary team and the patients. PMID- 15805027 TI - Modernizing medical photography, part 1. AB - Government, media and public focus on waiting times in the National Health Service in the United Kingdom has forced the organization to look closely at the process by which a patient progresses through an increasingly complex and ever changing system. In an effort to streamline the patient journey or care pathway, modernizers have turned to business and manufacturing for solutions. Whilst medical photographers need to recognize their role in this context, they are also facing major technological modernization through the development of digital photography. Part 1 of this paper looks at the origins of some of the techniques presently being used to modernize the patient journey. Part 2 shows how these tools of modernization can be utilized to harness the advantages of digital technology to provide a modern and appropriate medical photography service in a large, disparate teaching hospital. PMID- 15805029 TI - Photography of cleft audit patients. AB - The IMI National Guidelines have been prepared as baseline guides on specific aspects of medical illustration practice, and provide auditable standards for the future. They can be implemented in full, or may be amended according to individual requirements. The following, which is the first in the series to be published in this journal, is an abridged version of guidelines prepared by Marie Jones. The full version of this and the other guidelines can be read or downloaded at www.imi.org.uk. PMID- 15805045 TI - Polybrominated diphenyl ethers (PBDEs) in U.S. computers and domestic carpet vacuuming: possible sources of human exposure. AB - Polybrominated diphenyl ethers (PBDEs), a type of brominated flame retardant chemically and toxicologically similar to polychlorinated biphenyls (PCBs), are a class of emerging environmental and human contaminants. They have recently been detected in U.S. milk, blood, and food at the highest levels in the world. This pilot study was undertaken with the aim of determining levels of PBDE in the U.S. indoor environment, to assess the potential exposure to PBDEs from computer surfaces and carpets. Food of animal origin is the usual source of polychlorinated dibenzo-p-dioxin (PCDD), polychlorinated dibenzofurans (PCDF), and PCBs in humans, but there may also be environmental sources for intake of PBDEs. It was also our aim to characterize the PBDE congener profile in these indoor environmental samples. Four computer wipe samples and 9 domestic vacuum sweeping samples were analyzed for 13 PBDE congeners, PBDEs 17 (2,2',4), 28 (2,4,4'?), 47 (2,2',4,4'?), 66 (2,3',4,4'?), 77 (3,3',4,4'?), 85 (2,2',3,4,4'?), 99 (2,2'4,4',5), 100 (2,2',4,4',6), 138 (2,2',3,4,4',5'?), 153 (2,2',4,4',5,5'?), 154 (2,2',4,4',5,6'?), 183 (2,2',3,4,4',5',6), and 209 (2,2',3,3',4,4',5,5',6,6'?). All samples tested positive for PBDEs. PBDE 209 was the dominant congener in all 4 computer wipe samples and in 7 out of the 9 vacuum dust samples. The congener profiles observed in this study varied considerably, a finding that has been observed previously. However these congener profiles differ from the pattern seen in U.S. human milk, human blood and in food, where PBDEs 47 and 99 predominate. PMID- 15805046 TI - Effects of pentabrominated diphenyl ether (PBDE-99) on vitamin status in domestic duck (Anas platyrhynchos) hatchlings. AB - Polybrominated diphenyl ethers (PBDEs) used to prevent fire are found in several products, such as textiles, electronics, and building materials. They are lipophilic and persistent substances, and their toxicological endpoints resemble those of polychlorinated bipenyls (PCBs). One of the most abundant congeners of PBDEs in wildlife is the 2,2',4,4',5-pentabromodiphenyl ether (PBDE-99). To study the effect of PBDE-99 on vitamin status in birds and evaluate the possible application of vitamins as biomarkers for use in monitoring of wildlife, eggs of domestic duck (Anas platyrhynchos) were exposed by yolk sac injection to environmental realistic doses of this congener (0.1, 1, or 10 ng/g ww). Levels of retinol (vitamin A), retinyl palmitate, and a-tocopherol (vitamin E) were measured in the liver of the newly hatched chicks, and levels of retinol and tocopherol were also measured in plasma. Liver tocopherol levels correlated negatively to the exposure to PBDE-99. This is an indication that exposure to PBDE-99 reduces levels of tocopherol in liver. Thus, tocopherol seems to be a potential useful biomarker for exposure to PBDEs in bird species. PMID- 15805047 TI - Mercury impairment of mouse thymocyte survival in vitro: involvement of cellular thiols. AB - Heavy metals are well known to be able to induce immunotoxicity, but comparative metal studies related to apoptosis have not been conducted. In the present study, the effects of arsenic, cadmium, gold, lead, manganese, and mercury on thymocytes from BALB/c mice were analyzed. Thymic cells were cultured for 3-24 h in vitro in the absence or presence of metal, and markers of apoptosis or cell death, including annexin V binding, DNA loss/oligonucleosomal fragmentation, 7-amino actinomycin D uptake (loss of impermeance), changes of the mitochondrial membrane potential (JC-1 fluorescence), and Western analysis of cellular thiols, were assayed. Mercury (Hg) was the only metal shown to be consistently toxic with the dose and times utilized. Cadmium (Cd) was the only other metal tested that also produced some significant level of DNA loss; however, the induction of apoptosis by Cd was not as consistent as that observed with Hg. When Hg was added with 2 mercaptoethanol (2-ME), Hg produced greater toxicity. Endogenous DNA synthesis by thymocytes was immediately inhibited by Hg and Hg + 2-ME. The Hg + 2-ME-induced apoptosis appeared to be associated with altered levels of cellular thiols, in that glutathione (GSH) depletion was significant in comparison to the non-metal control and Hg alone. The increased Hg-induced toxicity in the presence of 2-ME likely was due to the ability of 2-ME to enhance (10- to 20-fold) the cellular uptake of Hg. Western analysis with biotin maleimide demonstrated that Hg + 2-ME and to a lesser extent the positive control dexamethasone eliminated many reactive thiols; the major thiol-reactive protein still reactive with the maleimide probe had an approximate Molecular Mass of 45 kD. Surprisingly, Hg alone enhanced the expression of this thiol-expressing protein, which by Mass Spectrometry (MS)/MS analysis was shown to be beta-actin. Hg also produced the appearance of yet to be identified new proteins. Based on the results with Hg + 2 ME, it is suggested that numerous protein thiols participate in maintenance of cell survival and their loss is associated with apoptosis. The increased expression of new thiol-reactive proteins or thiol-reactive proteins with altered electrophoretic profiles needs to be further investigated. However, the enhanced toxicity attributed to Hg + 2-ME suggests that increased intracellular oxidative stress, observed as increased depletion of GSH, is responsible for the accelerated cell death. PMID- 15805048 TI - Effects of nonylphenol on rates of tail resorption and metamorphosis in Rana catesbeiana tadpoles. AB - Nonylphenol (NP) is a persistent, lipophilic, and toxic chemical that can be endocrine disrupting (estrogenic) at sublethal concentrations. Since amphibian metamorphosis is a hormone-driven process and a delicate balance of hormone levels is required for successful metamorphosis, exposure of larval amphibians to NP might disrupt metamorphic processes. This study tested whether NP exposure influenced rate of metamorphic progression and tail resorption in bullfrog (Rana catesbeiana) tadpoles. Premetamorphic bullfrog tadpoles were exposed for 7 d to one of 3 nominal concentrations of NP (234 microg/L, 468 microg/L, or 936 microg/L) with or without the addition of exogenous 3,3',5-triiodothyronine (T3). In the absence of exogenous T3, NP significantly increased the rate of tail growth (as measured by tail length) at 936 microg/L. There was no significant effect of NP alone on tail width, limb development, or the process of cranial transformation. When T3 was added to the treatments, increasing NP concentrations were associated with a significant decrease in the rate of cranial transformation, and at the highest dose, the rate of tail resorption was significantly lower than in the controls. Overall, NP had an inhibitory effect on the rate of bullfrog tadpole metamorphic progression and tail resorption. PMID- 15805049 TI - Idiosyncratic liver injury: challenges and approaches. AB - Clinical presentations of drug-induced liver injury (DILI) cover essentially the entire spectrum of known liver diseases. However, in the last 8 years the form of liver injury that has most frequently resulted in labeling restrictions is idiosyncratic hepatocellular injury leading to acute liver failure. This rare form of DILI has a characteristic clinical presentation that includes an acute onset after uneventful treatment with drug for weeks to months. Serum alanine aminotransferase rises to very high levels and the appearance of jaundice indicates a high mortality even if the therapy is discontinued. Drugs that can cause this type of injury almost always are associated with frequent (2-15% of all treated patients) and minor serum aminotransferase elevations. These elevations are believed to reflect true liver injury, but often reverse even if drug therapy is continued. The bases for this "adaptation" is not known, as is why some patients do not adapt and develop progressive liver injury. Understanding how drugs cause severe idiosyncratic hepatocellular toxicity has been frustrated by the lack of good preclinical models. Indeed, because these events occur so rarely, the vast majority of humans are not good models. Studies of genomic DNA from affected individuals should provide important insight but not the complete answer because environmental factors almost certainly contribute to individual susceptibility. The most fruitful approach may therefore lie in focused and well-controlled phenotype/genotype studies of the rare patients who have survived this type of injury. The National Institute of Diabetes and Digestive and Kidney Diseases of The National Institutes of Health has recently sponsored a cooperative agreement (UO1) to create a Drug Induced Liver Injury Network (DILIN). DILIN consists of University of Michigan, Indiana University, University of Connecticut, University of California, San Francisco, University of North Carolina, and Duke University. This network should provide heretofore missing resources required to address the problem. PMID- 15805050 TI - Mechanistic classification of liver injury. AB - Toxic injury occurs in the liver more often than any other organ. This can be attributed to the fact that virtually all ingested substances that are absorbed are first presented to the liver and that the liver is responsible for the metabolism and elimination of many substances. Drug-induced liver injury has become a serious health problem in contemporary society. Moreover, liver toxicity is a significant impediment to development of new pharmaceuticals. A classification of liver injury is presented as a means to better understand the spectrum of known mechanisms of liver injury and to assist in discovering novel pathways of toxic liver injury. PMID- 15805051 TI - Clinical pathology approaches to hepatic injury. AB - Testing the blood for evidence of hepatic damage and dysfunction frequently involves measuring several blood constituents simultaneously to screen for disease. While useful, this approach occasionally leads to apparent disparities between the blood test results, and the results of other diagnostic tests such as histology. In part, these perceived discrepancies may stem from a lack of appreciation for tissue, cellular, and molecular factors that affect the appearance of hepatic disease biomarkers in the blood. Further confusing the matter is that in some instances the mechanisms responsible for the appearance of diagnostic compounds in blood are only partially understood. Many of the known factors that affect hepatic biomarkers are similar to those affecting other tissue markers, while others are unique to the liver, such as those involved with cholestasis. Disease conditions can also cause misleading results by affecting tissue concentrations of test compounds, hepatic mass, and the clearance rate of compounds from the blood. Knowledge of the factors affecting the blood concentrations of biomarkers, as well as investigations into the mechanisms behind changes to hepatic biomarker concentrations, may allow for a better interpretation of blood test results and fewer inconsistencies between diagnostic results. PMID- 15805053 TI - New insights into functional aspects of liver morphology. AB - The liver is structurally and functionally complex and has been considered second only to brain in its complexity. Many mysteries still exist in this heterogeneous tissue whose functional unit of the lobule has continued to stump morphologists for over 300 years. The primary lobule, proposed by Matsumoto in 1979, has been gaining acceptance as the functional unit of the liver over other conceptual views because it's based on vessel architecture and includes the classic lobule as a secondary feature. Although hepatocytes comprise almost 80% of the liver, there are at least another dozen cell types, many of which provide "cross-talk" and play important functional roles in the normal and diseased liver. The distribution and functional roles of all cells in the liver must be carefully considered in both the analysis and interpretation of research data, particularly data in the area of genomics and "phenotypic anchoring" of gene expression results. Discoveries regarding the functional heterogeneity of the various liver cell types, including hepatocytes, hepatic stellate cells, sinusoidal endothelia, and Kupffer cells, are providing new insights into our understanding of the development, prevention and treatment of liver disease. For example, functional differences along zonal patterns (centrilobular or periportal) have been demonstrated for sinusoidal endothelium, Kupffer cells, and hepatocytes and can explain the gradients and manifestations of disease observed within lobules. Intralobular gradients of bile uptake, glycogen depletion, glutamine synthetase, and carboxylesterase by hepatocytes; widened fenestrations in centrilobular sinusoidal lining cells; and differences in the components of centrilobular extracellular matrix or function of Kupffer cells have been demonstrated. Awareness of the complexities and heterogeneity of the liver will add to a greater understanding of liver function and disease processes that lead to toxicity, cancer, and other diseases. PMID- 15805052 TI - New technologies and screening strategies for hepatotoxicity: use of in vitro models. AB - Hepatotoxicity remains a significant cause for drug failures during clinical trials. This is due, in part, to the idiosyncratic nature of toxicity in humans and inherent physiological differences between humans and preclinical species leading to limited correct prediction of adverse responses in humans. To address this issue, robust screening assays are being developed, which have heightened predictive capacity for human hepatotoxicity, and may be utilized throughout the discovery and development phases in conjunction with traditional in vivo methods, for decision making during drug selection and risk assessment. This manuscript describes an example application of in vitro-based strategies using human hepatocyte cultures in lead optimization screening in conjunction with ADME profiling, for evaluation of compound-associated CYP450 induction potential, and the identification of potentially useful biomarkers as predictors of hepatotoxicity for use in vitro, and in preclinical species and humans. PMID- 15805054 TI - Comparative hepatic toxicity: prechronic/chronic liver toxicity in rodents. AB - The morphologic assessment of the gross and microscopic appearance of the liver can provide a broad base of knowledge concerning the potential toxicity of a drug or chemical. This information may either lead to an understanding of the underlying mechanism of toxicity or guide further study to discern the mode of action of the hepatotoxicity. In standard regulatory bioassays, toxicity studies are conducted during phase 1 and phase 2 of the development process to define the acute, subchronic and chronic toxicity of the test compound. In the liver, there are a limited number of morphologic changes that can be identified using conventional light microscopy. These morphologic alterations are often characterized as "adaptive," consisting of an exaggerated normal physiologic response; "pharmacologic," consisting of an expected alteration in response to the desired action of the test article; or "adverse," consisting of morphologic alterations that are generally undesired, progressive and deleterious to the normal function of the cell(s) involved. Morphologic evidence of adverse effects may involve hepatocytes, the biliary system, hepatic vasculature, Kupffer cells, or stellate cells (Ito cells). In drug discovery and development programs, it is necessary to utilize a multidisciplinary approach, using different endpoints, to investigate the same or similar biological responses in the liver. This results in large amounts of data that must be organized in a retrievable fashion. In order for such a multidisciplinary approach to succeed, each discipline must organize and generate their data in a manner that is easily used by others in the process. The toxicologic pathologist must develop and use standardized nomenclature and diagnostic criteria when examining the liver so that data from various investigators can be compared in a useful manner. PMID- 15805055 TI - Tissue repair: an important determinant of final outcome of toxicant-induced injury. AB - Tissue repair is a dynamic compensatory cell proliferation and tissue regeneration response stimulated in order to overcome acute toxicity and recover organ/tissue structure and function. Extensive evidence in rodent models using structurally and mechanistically diverse hepatotoxicants such as acetaminophen (APAP), carbon tetrachloride (CCl4), chloroform (CHCl3), thioacetamide (TA), trichloroethylene (TCE), and allyl alcohol (AA) have demonstrated that tissue repair plays a critical role in determining the final outcome of toxicity, i.e., recovery from injury and survival or progression of injury leading to liver failure and death. Tissue repair is a complex process governed by intricate cellular signaling involving a number of chemokines, cytokines, growth factors, and nuclear receptors leading to promitogenic gene expression and cell division. Tissue repair also encompasses regeneration of hepatic extracellular matrix and angiogenesis, the processes necessary to completely restore the structure and function of the liver tissue lost to toxicant-induced initiation followed by progression of injury. New insights have emerged over the last quarter century indicating that tissue repair follows a dose response. Tissue repair increases with dose until a threshold dose, beyond which it is delayed and impaired due to inhibition of cellular signaling resulting in runaway secondary events causing tissue destruction, organ failure, and death. Prompt and adequately stimulated tissue repair response to toxic injury is critical for recovery from toxic injury. Tissue repair is modulated by a variety of factors including species, strain, age, nutrition, and disease condition causing marked changes in susceptibility and toxic outcome. This review focuses on the properties of tissue repair, different factors affecting tissue repair, and the mechanisms that govern tissue repair and progression of injury. It also highlights the significance of tissue repair as a target for drug development strategies and an important consideration in the assessment of risk from exposure to toxicants. PMID- 15805056 TI - Liver immunobiology. AB - Tle liver has a number of important functions in innate and adaptive immunity. Contributions to the innate (nonspecific) immune system include production of acute phase proteins, nonspecific phagocytosis of particles, nonspecific pinocytosis of molecules, and nonspecific cell killing. Hepatic involvement in innate immunity contributes to the systemic response to local inflammation, clearance of particles and soluble molecules from the circulation, and killing of invading cells such as neoplastic cells. Liver involvement in the adaptive (specific) immune system includes deletion of activated T cells, induction of tolerance to ingested and self-antigens, extrathymic proliferation of T cells, and deletion of many of the signaling and effector molecules. Hepatic involvement in adaptive immunity allows clearance of activated T cells and signaling molecules following inflammatory reactions, and promotes immunologic tolerance toward potentially antigenic proteins that are absorbed from the intestinal tract. The liver is a major site of extrathymic T cell development, which assumes increasing significance with aging in mammals. Perturbations in hepatic structure or function can result in significant ramifications in both the innate and adaptive immune systems. PMID- 15805057 TI - Spontaneous and drug-induced hepatic pathology of the laboratory beagle dog, the cynomolgus macaque and the marmoset. AB - This review focuses on the background hepatic pathology present in three of the most commonly used species in the safety assessment of drugs, namely the beagle dog, the marmoset and the cynomolgus macaque. Both the nonneoplastic and neoplastic pathology are reviewed with a discussion on the potential impact that significant background pathology might have on the interpretation of any drug induced pathology during subsequent testing. Although specific instances, such as parasitological infection in wild-caught primates can pose problems of interpretation, in general the background pathology in both the dog and the nonhuman primates, is not significantly different from that seen in the liver of laboratory rodents and with experience should not pose significant problems for the experienced pathologist. The relative merits of the primate versus the dog as a choice of second species are also considered in some detail. Although there is an inbuilt prejudice that the primate will more closely mimic subsequent effects that might occur in man in the clinic, insofar as the liver is concerned, there are many instances where the dog has been more representative of human exposure and metabolism and there is little evidence to show that the nonhuman primate is consistently better than dog in predicting human liver toxicity. As with most areas of science, comparative toxicology would dictate that the more information gained, from as wide a range of species as is practical, will give the best assessment for any subsequent problems in the clinic. This pragmatic approach should prove to be more successful than one based entirely upon an assumption, and in many cases the assumption is incorrect, that the primate always predicts human toxicity better than the nonprimate, including the dog. PMID- 15805058 TI - A brief overview of nonneoplastic hepatic toxicity in fish. AB - Biochemical assays are not routinely used to assess liver damage in fish, therefore, a histopathological evaluation is usually required to determine the existence or extent of nonneoplastic liver toxicity. Many mammalian pathologists may be uncomfortable when requested to identify and interpret subtle liver changes in these unfamiliar animals. It may be reassuring to note that there are more similarities than differences between fish and mammals in terms of their macro- and microanatomy, physiological and biochemical characteristics, and pathologic responses to hepatotoxic substances. This brief overview addresses several topics pertaining to hepatotoxicity in fish, including: anatomic considerations, that is, how the anatomy of the fish liver may be predictive of its metabolic capacity, and also its microscopic appearance, following exposure to toxins; physiologic considerations, including comparisons between mammalian and fish livers regarding the uptake, elimination, toxification, or detoxification of xenobiotic compounds; morphologic responses to toxicity, in which some of the general types of findings that are most commonly observed in cases or studies of fish hepatotoxicity are highlighted; and last, responses of the fish liver to specific hepatotoxins. PMID- 15805059 TI - Using laser scanning cytometry to measure PPAR-mediated peroxisome proliferation and beta oxidation. AB - Laser scanning cytometry (LSC) is a new technology that combines the properties and advantages of flow cytometry (FC) and immunohistochemistry (IHC), thus providing qualitative and quantitative information on protein expression with the additional perspective provided by cell and tissue localization. Formalin-fixed, paraffin embedded liver sections from rats exposed to a Peroxisome Proliferator Activated Receptor (PPAR) agonist were stained with antibodies against peroxisomal targeting signal-1 (PTS-1) (a highly conserved tripeptide contained within all peroxisomal enzymes), Acyl CoA oxidase (AOX) (the rate limiting enzyme of peroxisomal beta oxidation), and catalase (an inducible peroxisomal antioxidant enzyme) to evaluate peroxisomal beta oxidation, oxidative stress, and peroxisome proliferation. The LSC showed increased AOX, catalase, and PTS-1 expression in centrilobular hepatocytes that correlated favorably with the microscopic observation of centrilobular hepatocellular hypertrophy and with the palmitoyl CoA biochemical assay for peroxisomal beta oxidation, and provided additional morphologic information about peroxisome proliferation and tissue patterns of activation. Therefore, the LSC provides qualitative and quantitative evaluation of peroxisome activity with similar sensitivity but higher throughput than the traditional biochemical methods. The additional benefits of the LSC include the direct correlation between histopathologic observations and peroxisomal alterations and the potential utilization of archived formalin-fixed tissues from a variety of organs and species. PMID- 15805061 TI - Variation in the hepatic gene expression in individual male Fischer rats. AB - A new tool beginning to have wider application in toxicology studies is transcript profiling using microarrays. Microarrays provide an opportunity to directly compare transcript populations in the tissues of chemical-exposed and unexposed animals. While several studies have addressed variation between microarray platforms and between different laboratories, much less effort has been directed toward individual animal differences especially among control animals where RNA samples are usually pooled. Estimation of the variation in gene expression in tissues from untreated animals is essential for the recognition and interpretation of subtle changes associated with chemical exposure. In this study hepatic gene expression as well as standard toxicological parameters were evaluated in 24 rats receiving vehicle only in 2 independent experiments. Unsupervised clustering demonstrated some individual variation but supervised clustering suggested that differentially expressed genes were generally random. The level of hepatic gene expression under carefully controlled study conditions is less than 1.5-fold for most genes. The impact of individual animal variability on microarray data can be minimized through experimental design. PMID- 15805060 TI - Imaging mass spectrometry: principles and potentials. AB - Direct tissue profiling and imaging mass spectrometry (MS) allow for detailed mapping of the complex protein pattern across a tissue sample. Utilization of these tools provides spatial information across a tissue section for target protein expression and can be used to correlate changes in expression levels with specific disease states or drug response. Protein patterns can be directly correlated to known histological regions within the tissue, allowing for the direct monitoring of proteins specific for morphological regions within a tissue sample. Profiling and imaging MS have been used to characterize multiple tissues, including human gliomas and lung cancers, as well as tumor response to specific therapeutics, suggesting the use of proteomic information in assessing disease progression as well as predicting patient response to specific treatments. This article discusses both the technology and methods involved in analyzing proteins directly from tissue samples as well as several MS applications, including profiling human tumors, characterizing protein differences between tumor grades, and monitoring protein changes due to drug therapy. PMID- 15805063 TI - Differential display in rat livers treated for 13 weeks with phenobarbital implicates a role for metabolic and oxidative stress in nongenotoxic carcinogenicity. AB - Hepatic enzyme inducers such as phenobarbital are often nongenotoxic rodent hepatocarcinogens. Currently, nongenotoxic hepatocarcinogens can only be definitively identified through costly and extensive long-term, repeat-dose studies (e.g., 2-year rodent carcinogenicity assays). Although liver tumors caused by these compounds are often not found to be relevant to human health, the mechanism(s) by which they cause carcinogenesis are not well understood. Toxicogenomic technologies represent a new approach to understanding the molecular bases of toxicological liabilities such asnongenotoxic carcinogenicity early in the drug discovery/development process. Microarrays have been used to identify mechanistic molecular markers of nongenotoxic rodent hepatocarcinogenesis in short-term, repeat-dose preclinical safety studies. However, the initial "noise" of early adaptive changes may confound mechanistic interpretation of transcription profiling data from short-term studies, and the molecular processes triggered by treatment with a xenobiotic agent are likely to change over the course of long-term treatment. Here, we describe the use of a differential display technology to understand the molecular mechanisms related to 13 weeks of dosing with the prototype rodent nongenotoxic hepatocarcinogen, phenobarbital. These findings implicate a continuing role for oxidative stress in nongenotoxic carcinogenicity.An Excel data file containing raw data is available in full at http://taylorandfrancis.metapress.com/openurl.asp?genre=journal&issn=0192-6233. Click on the issue link for 33(1), then select this article. A download option appears at the bottom of this abstract. The file contains raw data for all gene changes detected by AFLP, including novel genes and genes of unknown function; sequences of detected genes; and animal body and liver weight ratios. In order to access the full article online, you must either have an individual subscription or a member subscription accessed through www.toxpath.org. PMID- 15805062 TI - Transcriptional profiling of the left and median liver lobes of male f344/n rats following exposure to acetaminophen. AB - The liver is a common organ for transcriptional profiling because of its role in xenobiotic metabolism and because hepatotoxicity is a common response to chemical exposure. To explore the impact that sampling different lobes may have on transcriptional profiling experiments we have examined and compared gene expression profiles of the left and median lobes of livers from male F344 rats exposed to toxic and nontoxic doses of acetaminophen. Transcript profiling using micorarrays revealed clear differences in the response of the left and median liver lobes of F344 rats to acetaminophen exposure both at low doses as well as doses that caused hepatotoxicity. Differences were found in the total number of differentially expressed genes in the left and median lobes, the number and identity of genes that were differentially expressed uniquely only in the left or median lobe, and in the patterns of gene expression. While it is not possible to generalize these results to compounds other than acetaminophen or other strains of rat, these results highlight the potential impact of sampling differences on the interpretation of gene expression profiles in the liver. PMID- 15805064 TI - Microarray data analysis of mouse neoplasia. AB - Microarray gene expression analysis offers great promise to help us understand the molecular events of experimental carcinogenesis, but have such promises been fulfilled? Studies of gene expression profiles of rodent are being published and demonstrate that yes, indeed, gene array data is furthering our understanding of tumor biology. Recent studies have identified differentially expressed genes in rodent mammary, colon, lung, and liver tumors. Although relatively few genes on the rodent arrays have been fully characterized, information has been generated to better identify signatures of histologic type and grade, understand invasion and metastasis, identify candidate biomarkers of early development, identify gene networks in carcinogenesis, understand responses to therapy, and decifer overlap with molecular events in human cancers. Data from mouse lung, mammary gland, and liver tumor studies are reviewed as examples of how to approach and interpret gene array data. Methods of gene array data analysis were also applied for discovery of genes involved in the regression of mouse liver tumors induced by chlordane, a nongenotoxic murine hepatocarcinogen. Promises are beginning to be fulfilled and it is clear that pathologists and toxicologists, in collaboration with molecular biologists, bioinformatists,and other scientists are making great strides in the design, analysis, and interpretation of microarray data for cancer studies. PMID- 15805065 TI - The hepatic transcriptome as a window on whole-body physiology and pathophysiology. AB - Transcriptomics can be a valuable aid to pathologists. The information derived from microarray studies may soon include the entire transcriptomes of most cell types, tissues and organs for the major species used for toxicology and human disease risk assessment. Gene expression changes observed in such studies relate to every aspect of normal physiology and pathophysiology. When interpreting such data, one is forced to look "far from the lamp post:' and in so doing, face one's ignorance of many areas of biology. The central role of the liver in toxicology, as well as in many aspects of whole-body physiology, makes the hepatic transcriptome an excellent place to start your studies. This article provides data that reveals the effects of fasting and circadian rhythm on the rat hepatic transcriptome, both of which need to be kept in mind when interpreting large scale gene expression in the liver. Once you become comfortable with evaluating mRNA expression profiles and learn to correlate these data with your clinical and morphological observations, you may wonder why you did not start your studies of transcriptomics sooner. Additional study data can be viewed at the journal website at (www.toxpath.org). Two data files are provided in Excel format, which contain the control animal data from each of the studies referred to in the text,including normalized signal intensity data for each animal (n=5) in the 6 hour, 24-hour, and 5-day time points. These files are briefly described in the associated 'Readme' file, and the complete list of GenBank numbers and Affymetrix IDs are provided in a separate txt file. These files are available at http://taylorandfrancis.metapress.comlopenurl.asp?genre=journal&issn=0192-6233. Click on the issue link for 33(1), then select this article. A download option appears at the bottom of this abstract. In order to access the full article online, you must either have an individual subscription or a member subscription accessed through (www.toxpath.org). PMID- 15805066 TI - Do preclinical testing strategies help predict human hepatotoxic potentials? AB - Overt hepatotoxicity due to drug administration is a real and present issue in drug development and regulatory circles. Preclinical drug development is intended to identify potential risks and target tissues prior to introduction of new molecular entities into the human population. The standard regimen is testing at various multiples of the intended human therapeutic dose in at least 2 species of animals, one rodent (rats or mice), one non-rodent (dogs,nonhuman primates, minipigs, and rabbits, as examples) for at least two weeks of repeated dosing. Experience has shown that this regimen "works"most of the time. However, preclinical models are not infallible and are not always predictive. Whether the lack of predictivity is due to individual human genetic sensitivities, immunologically mediated phenomena, disease mediation or idiosyncratic reactions, the animal models are limited in detecting these characteristics and other low incidence phenomena. While it is uncommon for drug developers to continue development with products that elicit overt hepatic toxicity early in the animal testing, some products have made it through the approval process and then shown significant adverse effects. Some of the drugs (acetaminophen, isoniazid, trovafloxacin, troglitazone, bromfenac, clarithromycin, telithromycin) that have shown this propensity will be discussed in detail from early preclinical development to marketing and, in some instances, to limitations to usage or removal from the U.S. marketplace. PMID- 15805067 TI - Recognizing drug-induced liver injury: current problems, possible solutions. AB - Currently there are three major problems in understanding drug-induced liver injury (DILI): (1) reliably establishing whether the liver disease was caused by the drug, or by another process; (2) determining the true incidence of and clinical risk factors for drug-induced hepatotoxicity; and(3) elaborating the mechanisms by which injury occurs to hepatocytes and other liver cells. We have focused here on the first two problems, as issues that may be amenable to actions in the near future, but the third may take substantially longer to work out. The first problem requires sufficient information for medical differential diagnosis. There are no pathognomonic indicators of DILI; even liver biopsy is not diagnostic. Making the correct attribution of causality requires analyzing the temporal relationship of drug exposure to illness and excluding all other possible causes. The second problem, determining incidence, cannot be done entirely adequately using currently available methods, whether by clinical trials, by spontaneous adverse event reports, or by retrospective epidemiologic studies. There is need for prospective safety studies to establish the true incidence of DILI caused by a drug, to identify risk factors for it, and to collect biologic materials for analytic studies toward better understanding mechanisms of DILI. PMID- 15805068 TI - Classification of proliferative hepatocellular lesions in harlan sprague-dawley rats chronically exposed to dioxin-like compounds. AB - Over the years, the most appropriate classification scheme for nodular proliferative lesions of the hepatocyte has been heavily debated. In the most recent guidelines there appears to be a consensus for classifying these lesions as hepatocellular adenoma, hepatocellular carcinoma, or regenerative hyperplasia. Also, large foci of cellular alteration may appear somewhat nodular. Some nodular hepatocellular lesions from a group of 7 studies of dioxin and dioxin-like compounds conducted by the National Toxicology Program did not readily fit into these categories. Some of these lesions had morphologic features consistent with hyperplasia. However, there was not sufficient morphological or biological evidence to conclude that the entire response was regenerative. In other instances, these lesions had some features resembling adenoma, but contained a prominent component of biliary epithelium and/or oval cells. This component does not appear to be well described in the literature, and while its presence suggested a nodule to be nonneoplastic, this is inconclusive. This paper describes the morphology of these lesions, as well as the diagnostic approach taken in this series of studies. PMID- 15805069 TI - Overview of the molecular biology of hepatocellular neoplasms and hepatoblastomas of the mouse liver. AB - The molecular pathogenesis of chemically induced hepatocellular neoplasms and hepatoblastomas in the B6C3FI mouse is unclear but may involve alterations in the fi-catenin/Wnt signaling pathway as was recently described for human liver neoplasms. The objectives of this research were to characterize the mutation frequency and spectrum of P-catenin mutations and the intracellular localization of I-catenin protein accumulation in chemically induced hepatoblastomas and hepatocellular neoplasms. In the majority of the hepatoblastomas examined by immunohistochemical methods, both nuclear and cytoplasmic localization of P catenin protein were detected, whereas in hepatocellular adenomas and carcinomas and normal liver only membrane staining was observed. Genomic DNA was isolated from paraffin sections of each liver tumor. P-catenin exon 2 (corresponds to exon 3 in humans) genetic alterations were identified in the majority of hepatoblastomas from exposed mice. Deletion mutations were identified more frequently than point mutations in hepatoblastomas. Hepatocellular adenomas and carcinomas from treated mice had mutations in exon 2 of the B-catenin gene which ranged from 32-43%, while 10% P-catenin mutations were detected in spontaneous neoplasms. By immunohistochemical methods cyclin Dl was observed in most nuclei of hepatoblastomas and strong expression of cyclin Dl was confirmed by Western analysis regardless of treatment. The cumulative data suggests that P-catenin mutations with upregulation of the B-catenin protein and Wnt signaling most likely increased cyclin Dl expression. Cyclin D1 may provide an advantage during tumor progression of hepatocellular neoplasms and hepatoblastomas. The review will also focus on other genes which are important in mouse and human liver tumors. PMID- 15805070 TI - Molecular mechanisms of hepatocarcinogenesis in transgenic mouse models of liver cancer. AB - Overexpression of c-myc and transforming growth factor-alpha (TGF-alpha) has been frequently observed in human hepatocellular carcinoma (HCC),suggesting a pivotal role played by these protooncogenes in liver oncogenesis. In order to investigate the molecular events underlying human hepatic malignant transformation, we have generated c-myc and c-myc/ TGF-alpha transgenic mice that are prone to liver cancer. These transgenic mice develop HCCs with different incidence, kinetics and histopathological features. Indeed, co-expression of c-myc and TGF-alpha transgenes results in a dramatic synergistic effect on liver tumor development when compared with respective single transgenic lines, including a shorter latency period and a more aggressive phenotype. The more malignant histopathological features characteristic of c-myc/ TGF-alpha HCCs are the result of the increased proliferation and reduced apoptosis in this model of liver cancer when compared with single parental lines. Accordingly, c-myc and c-myc/l TGF-alpha transgenic mice display a different molecular pathogenesis of HCC. Importantly, the genetic and molecular mechanisms that are involved in c-myc and c-myc/ TGF-alpha liver cancer development are major oncogenic events in human hepatocarcinogenesis, indicating that these mouse models represent a useful tool to dissect and elucidate the molecular basis of human HCC. PMID- 15805073 TI - Modulation of oral squamous cell carcinoma incidence in rats via diet and a novel calcium channel antagonist. AB - An unexpected dose related increase in oral squamous cell carcinomas was observed in a standard 2-year carcinogenicity study with a novel calcium channel blocker, in which Wistar rats received daily doses of 0, 1.5, 7, 20, or 40 mg/kg of the compound mixed with a standard diet containing fibers from barley. This finding was associated with an increased incidence of severe (destructive) periodontitis and the formation of oro-nasal fistulae at the 2 highest doses. Five assays of the compound for genotoxicity were negative indicating that a genotoxic effect was highly improbable. To investigate the underlying pathogenic mechanisms a second 2-year study in the same strain of rats was initiated and the influence of the diet and/or a possible local irritancy by the drug was assessed. In this second study the compound was administered by oral gavage at daily doses of 0, 7, or 40 mg/kg (later reduced to 20 mg/kg due to systemic intolerance) to rats maintained either on the standard diet or on a low fiber diet assumed to be less aggressive in terms of inducing periodontal lesions. Dose dependent gingival overgrowth (a class-related effect) was observed in the incisor and molar teeth area of all treated groups but was independent of the diet used. No oral tumors were found in the standard diet or low fiber diet controls and all treatment groups fed the low fiber diet, whereas in the high-dose group fed the standard diet a total of 8 oral squamous cell carcinomas were detected in association with an increased incidence of severe periodontitis. These results indicate that the increased incidence of squamous cell carcinomas observed upon chronic administration of the compound is not due to a direct tumorigenic effect of the drug. Tumor formation is attributable to severe periodontal disease favored by the diet and class related gingival overgrowth. PMID- 15805072 TI - Prediction of nephrotoxicant action and identification of candidate toxicity related biomarkers. AB - A vast majority of pharmacological compounds and their metabolites are excreted via the urine, and within the complex structure of the kidney,the proximal tubules are a main target site of nephrotoxic compounds. We used the model nephrotoxicants mercuric chloride, 2-bromoethylamine hydrobromide, hexachlorobutadiene, mitomycin, amphotericin, and puromycin to elucidate time- and dose-dependent global gene expression changes associated with proximal tubular toxicity. Male Sprague-Dawley rats were dosed via intraperitoneal injection once daily for mercuric chloride and amphotericin (up to 7 doses), while a single dose was given for all other compounds. Animals were exposed to 2 different doses of these compounds and kidney tissues were collected on day 1, 3, and 7 postdosing. Gene expression profiles were generated from kidney RNA using 17K rat cDNA dual dye microarray and analyzed in conjunction with histopathology. Analysis of gene expression profiles showed that the profiles clustered based on similarities in the severity and type of pathology of individual animals. Further, the expression changes were indicative of tubular toxicity showing hallmarks of tubular degeneration/regeneration and necrosis. Use of gene expression data in predicting the type of nephrotoxicity was then tested with a support vector machine (SVM)-based approach. A SVM prediction module was trained using 120 profiles of total profiles divided into four classes based on the severity of pathology and clustering. Although mitomycin C and amphotericin B treatments did not cause toxicity, their expression profiles were included in the SVM prediction module to increase the sample size. Using this classifier, the SVM predicted the type of pathology of 28 test profiles with 100% selectivity and 82% sensitivity. These data indicate that valid predictions could be made based on gene expression changes from a small set of expression profiles. A set of potential biomarkers showing a time- and dose-response with respect to the progression of proximal tubular toxicity were identified. These include several transporters (Slc21a2, Slc15, Slc34a2), Kim 1, IGFbp-1, osteopontin, alpha fibrinogen, and Gstalpha. PMID- 15805075 TI - Postmortem hepatocyte vacuolation in cynomolgus monkeys. PMID- 15805074 TI - Proliferating cell nuclear antigen--a marker for ovarian follicle counts. AB - Enumerating ovarian follicles is an effective way to estimate the extent of ovarian toxicity in female rodents exposed to xenobiotics. Differential follicle counts are useful in safety assessment bioassays and in interspecies extrapolation of ovarian toxicity. Counting the follicles in H&E-stained sections is labor intensive, tedious, and costly. In the present study we demonstrated that in rat formalin-fixed, paraffin-embedded ovary sections follicles of all degrees of maturity can be visualized by the use of antibody directed against proliferating cell nuclear antigen (PCNA). Follicles are easily distinguished from ovarian background with the ability to detect and identify primordial follicles being enhanced. This translates into a significant decrease in variability of follicle counts, labor, and cost. Specifically, variability dropped from 11% to 0.2%, the counting time was reduced by 46%, and the cost by 48%. PMID- 15805076 TI - Olfactory epithelial metaplasia and hyperplasia in female Harlan Sprague-Dawley rats following chronic treatment with polychlorinated biphenyls. AB - The National Toxicology Program recently completed a series of studies to evaluate the relative potency for toxicity and carcinogenicity of several polyhalogenated aromatic hydrocarbons including dioxin-like compounds (DLCs) and polychlorinated biphenyls. Female Sprague-Dawley rats were administered by gavage for up to 2 years with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD); 3,3',4,4',5 pentachlorobiphenyl (PCB126); 2,3,4,7,8-pentachlorodibenzofuran (PeCDF); 2,2',4,4',5,5'-hexachlorobiphenyl (PCB153); a tertiary mixture of TCDD, PCB126, and PeCDF; a binary mixture of PCB126 and 153; or a binary mixture of PCB126 and 2,3',4,4',5-pentachlorobiphenyl (PCB118); control animals received corn oil acetone vehicle (99:1) alone. Nasal epithelial changes were observed only in animals exposed for 2 years to the higher doses of the binary mixtures of PCB126 + PCB153 (1000 ng/kg and 1000 microg/kg) and PCB126 + PCB118 (216 and 360 ng TCDD equivalents/kg). In both studies, the changes were of the same nonneoplastic nature, localized to nasal sections II and III located, respectively, at the level of the incisive papilla anterior to the first palatial ridge (section II) and through the middle of the second molar teeth (section III). The changes consisted of hyperplasia of the respiratory epithelium (level II) and metaplasia of olfactory epithelium to respiratory epithelium with further hyperplasia of the metaplastic respiratory epithelium (levels II and III). Variable amounts of acute inflammatory exudate appeared within the lumen of the nasal cavity, overlying the affected epithelium. Occasionally, the inflammation eroded through the skull and into the adjacent olfactory bulbs. PMID- 15805077 TI - Neuropathological studies of rats following multiple exposure to tri-ortho-tolyl phosphate, chlorpyrifos and stress. AB - Adult male Long-Evans rats were exposed to 2 neurotoxic organophosphates in a setting of chronic stress, over a 63-day period. The organophosphates were tri ortho-tolyl phosphate (TOTP) administered in 14 gavage doses of 75, 150 or 300 mg/kg, and chlorpyrifos, given in two 60 mg/kg subcutaneous exposures. Corticosterone was added to the drinking water at 400 microg/ml, to model aspects of chronic stress. These compounds/dosages were administered individually and in combination, with appropriate controls, giving rise to 16 experimental groups. The major neuropathologic change was the presence of axonal degeneration progressing to myelinated fiber degeneration, mainly in distal regions of selected fiber tracts and peripheral nerve, seen in animals sacrificed on experimental day 63. The cervical spinal cord and medullary levels of the sensory gracile fasciculus were most prominently affected. This axonopathy/fiber degeneration was TOTP dose-related at the 300 and 150 mg/kg levels. There was association of this lesion with inhibition of the enzyme neurotoxic esterase in hippocampal tissue from TOTP-treated rats. Such an association categorizes this disease process as organophosphate ester-induced delayed neuropathy. Neither chlorpyrifos nor corticosterone appeared to contribute to the neuropathic events or the enzyme inhibition. A cohort of rats was maintained on the corticosterone dosing, but without additional exposure to TOTP or chlorpyrifos, for an additional 27 days. When these rats were examined on day 90, the nerve fiber degeneration had progressed in all experimental groups administered the 300 mg/kg dose of TOTP (lower doses were not studied at the 90-day interval), although hippocampal neurotoxic esterase had returned to control values. PMID- 15805078 TI - An investigation of the effects of late-onset dietary restriction on prostate cancer development in the TRAMP mouse. AB - In our previous work we showed that dietary restriction initiated at puberty reduced prostate cancer development in the TRAMP mouse model. The current study was conducted to ascertain whether a dietary restriction regime would similarly reduce lesion development if imposed once tumor development was well established. Male TRAMP mice were maintained on an ad libitum diet until 20 weeks of age when proliferative prostate lesions are clearly evident. Mice were then subjected to a 20% restriction in dietary calories compared to matched controls, which were continued on ad libitum feeding. Mice were sacrificed at 20, 24, 32, and 39 weeks of age and proliferative epithelial lesions of the prostate were assessed using an established grading scheme. In this study, although dietary restriction reduced mean sex pluck weight (prostate and seminal vesicles), and mean grade of epithelial proliferative lesions in the dorsal and lateral lobes of the prostate, the effect was not as pronounced as was the case with dietary restriction from puberty. There was no relationship between serum insulin like growth factor (IGF 1) and prostate lesion grade. Additionally, we also report the relationship between lobe specific lesion development and SV40 immunostaining and, the occurance of neuroendocrine tumors (NETs) in the ventral prostate and urethra of the TRAMP mouse. NETs stained with high specificity and sensitivity for the neuroendocrine markers, synaptophysin and neuron-specific enolase (NSE), less for serotonin, but not for chromogranin A. NETs did not stain for cyclo-oxygenase-2 (COX-2) nor androgen receptor (AR). SV40 positive tubulo-acinar tumors seen occasionally in the kidney, did not stain for synaptophysin nor NSE. PMID- 15805079 TI - Experimental renal failure and iron overload: a histomorphometric study in rat tibia. AB - Renal failure (RF) is a serious disease of relatively high incidence, known to cause bone alterations. RF patients frequently suffer anemia, which is usually treated with iron. Given that iron overload inhibits bone formation, the aim of the present study was to evaluate the effect of iron on the subchondral bone of rat tibiae, using a model of renal failure. Male Wistar rats were subjected to experimental nephrectomy in order to induce renal failure and to iron overload by daily intraperitoneal injections of 88 mg/kg body weight of iron-dextran for 16 days. Tetracyclines were injected intraperitoneally to evaluate dynamic parameters of bone. Undecalcified histological sections of the tibiae were obtained. Serum urea, creatinine, and paratohormone (PTH) levels were evaluated 30 days after the onset of the experiment. Static and dynamic histomorphometric measurements were performed. Iron overload modified the response of the animals with renal failure: a reduction in bone forming activity compatible with adynamic bone disease and a decrease in peritrabecular fibrosis were observed. Our results suggest that iron is yet one more factor involved in the imbalance in bone metabolism typically found in renal failure patients treated with iron, rendering diagnosis and treatment of bone disease in these patients more complex. PMID- 15805080 TI - STP position paper: best practice guideline for the routine pathology evaluation of the immune system. PMID- 15805082 TI - STP position paper: ovarian follicular counting in the assessment of rodent reproductive toxicity. PMID- 15805084 TI - Responses of deciduous forest trees to severe drought in Central Europe. AB - In 2003, Central Europe experienced the warmest summer on record combined with unusually low precipitation. We studied plant water relations and phenology in a 100-year- old mixed deciduous forest on a slope (no ground water table) near Basel using the Swiss Canopy Crane (SCC). The drought lasted from early June to mid September. We studied five deciduous tree species; half of the individuals were exposed to elevated CO(2) concentration ([CO(2)]) (530 ppm) using a free air, atmospheric CO(2)-enrichment system. In late July, after the first eight weeks of drought, mean predawn leaf water potential about 30 m above ground was 0.9 MPa across all trees, dropping to a mean of -1.5 MPa in mid-August when the top 1 m of the soil profile had no plant accessible moisture. Mean stomatal conductance and rates of maximum net photosynthesis decreased considerably in mid August across all species. However, daily peak values of sap flow remained surprisingly constant over the whole period in Quercus petraea (Matt.) Liebl., and decreased to only about half of the early summer maxima in Fagus sylvatica L. and Carpinus betulus L. (stomatal down- regulation of flux). Although we detected no differences in most parameters between CO(2)-treated and control trees, predawn leaf water potential tended to be less negative in trees exposed to elevated [CO(2)]. Leaf longevity was greater in 2003 compared with the previous years, but the seasonal increase in stem basal area reached only about 75% of that in previous years. Our data suggest that the investigated tree species, particularly Q. petraea, did not experience severe water stress. However, an increased frequency of such exceptionally dry summers may have a more serious impact than a single event and would give Q. petraea a competitive advantage in the long run. PMID- 15805085 TI - Intra-annual variations in climate influence growth and wood density of Norway spruce. AB - Intra-annual radial growth variations of two Norway spruce trees (Picea abies (L.) Karst.) were monitored over 4 years, at four heights up the stem, by means of point-dendrometers. The trees were then felled and radial wood samples were cut from the radii that had been monitored by the dendrometers and analyzed for density. From the radial growth measurements recorded by the dendrometers, we related positions within the rings to dates, thus making possible investigation of the relationships between changes within the rings in wood density and fluctuations in climate or growth rate. Radial growth started in early April and ended, with large intra-annual differences, in August or September. Short-term variations in growth rate were related to fluctuations in climate parameters and soil water reserves. The sensitivity of radial growth to climate decreased with stem height. Wood density responded strongly to drought events, and a dry period in June 1996 induced false-ring formation. Wood density was relatively independent of growth rate and climatic conditions during the first part of the growing season, but increased with decreasing radial growth rate later in the growing season. PMID- 15805086 TI - A set of primers for analyzing chloroplast DNA diversity in Citrus and related genera. AB - Chloroplast simple sequence repeat (cpSSR) markers in Citrus were developed and used to analyze chloroplast diversity of Citrus and closely related genera. Fourteen cpSSR primer pairs from the chloroplast genomes of tobacco (Nicotiana tabacum L.) and Arabidopsis were found useful for analyzing the Citrus chloroplast genome (cpDNA) and recoded with the prefix SPCC (SSR Primers for Citrus Chloroplast). Eleven of the 14 primer pairs revealed some degree of polymorphism among 34 genotypes of Citrus, Fortunella, Poncirus and some of their hybrids, with polymorphism information content (PIC) values ranging from 0.057 to 0.732, and 18 haplotypes were identified. The cpSSR data were analyzed with NTSYS pc software, and the genetic relationships suggested by the unweighted pair group method based on arithmetic means (UPGMA) dendrogram were congruent with previous taxonomic investigations: the results showed that all samples fell into seven major clusters, i.e., Citrus medica L., Poncirus, Fortunella, C. ichangensis Blanco, C. reticulata Swingle, C. aurantifolia (Christm.) Swingle and C. grandis (L.) Osbeck. The results of previous studies combined with our cpSSR analyses revealed that: (1) Calamondin (C. madurensis Swingle) is the result of hybridization between kumquat (Fortunella) and mandarin (C. reticulata), where kumquat acted as the female parent; (2) Ichang papeda (C. ichangensis) has a unique taxonomic status; and (3) although Bendiguangju mandarin (C. reticulata) and Satsuma mandarin (C. reticulata) are similar in fruit shape and leaf morphology, they have different maternal parents. Bendiguangju mandarin has the same cytoplasm as sweet orange (C. sinensis), whereas Satsuma mandarin has the cytoplasm of C. reticulata. Seventeen PCR products from SPCC1 and 21 from SPCC11 were cloned and sequenced. The results revealed that mononucleotide repeats as well as insertions and deletions of small segments of DNA were associated with SPCC1 polymorphism, whereas polymorphism generated by SPCC11 was essentially due to the variation in length of the mononucleotide repeats. PMID- 15805087 TI - Acidic mist reduces foliar membrane-associated calcium and impairs stomatal responsiveness in red spruce. AB - Acidic deposition can leach essential pools of calcium (Ca) directly from plant foliage. Because of the central role of Ca in environmental signal transduction, disruptions of labile foliar Ca pools could impair physiological responses to a variety of environmental stimuli and stressors. We investigated the possibility that acidic mist-induced depletion of membrane-associated Ca (mCa), which is one form of labile Ca, may alter stomatal responsiveness to water stress, a process known to include Ca in signal transduction cascades. Red spruce (Picea rubens Sarg.) seedlings were exposed to either pH 3.0 or pH 5.0 mist treatments for one growing season. Foliar nutrition was assessed following treatments, and declines in stomatal conductance and net photosynthesis were measured on current-year shoots following stem excision. Seedlings exposed to pH 3.0 acidic mist treatments had reduced mCa relative to the pH 5.0 treated seedlings. Seedlings subjected to the pH 3.0 acidic mist treatment exhibited impaired stomatal functions, including a smaller maximum aperture, slower closure and an increased lag time between stomatal closure and photosynthetic decline following experimental water stress. Delayed stomatal closure could undermine desiccation avoidance mechanisms. Previous work has demonstrated that acidic mist treatments deplete mCa in red spruce and impair cold tolerance, with similar effects in other species. The results we present provide further evidence that acidic mist induced mCa depletion may cause disruption of a broad range of plant stress responses. PMID- 15805088 TI - Effects of mass inoculation on induced oleoresin response in intensively managed loblolly pine. AB - Oleoresin flow is an important factor in the resistance of pines to attack by southern pine beetle, Dendroctonus frontalis Zimm., and its associated fungi. Abiotic factors, such as nutrient supply and water relations, have the potential to modify this plant-insect-fungus interaction; however, little is known of the effects of inoculation with beetle-associated fungi on oleoresin flow. We observed that constitutive and induced resin yield in loblolly pine, Pinus taeda L., were affected by either fungal inoculation (with the southern pine beetle associated fungus Ophiostoma minus (Hedgcock) H. & P. Sydow) or silvicultural treatment. The effects of mass wounding (400 wounds m(-2)) and mass wounding and inoculation with O. minus were assessed by comparison with untreated (control) trees. The treatments were applied to trees in a 2 x 2 factorial combination of fertilizer and irrigation treatments. Fertilization did not significantly affect constitutive resin yield. Even as long as 105 days post-treatment, however, mass inoculated trees produced higher induced resin yields than control or wounded only trees, indicating a localized induced response to fungal inoculation. We noted no systemic induction of host defenses against fungal colonization. Although beetles attacking previously attacked trees face a greater resinous response from their host than beetles attacking trees that had not been previously attacked, the effect of an earlier attack may not last more than one flight season. Despite mass inoculations, O. minus did not kill the host trees, suggesting that this fungus is not a virulent plant pathogen. PMID- 15805089 TI - Decreasing photosynthesis at different spatial scales during the late growing season on a boreal cutover. AB - The relationship between photosynthesis and accumulated cold degree days (CDD) over the late growing season was examined at the shoot, ecosystem and landscape scales in a boreal cutover in eastern Canada predominated by black spruce (Picea mariana Mill. BSP), lowbush blueberry (Vaccinium angustifolium Ait.) and sheep laurel (Kalmia angustifolia L.). We calculated CDD as the sum of minimum daily temperatures below a 5 degrees C threshold. Light-saturated photosynthesis at the shoot level (A(max)) of black spruce and V. angustifolium decreased steadily with increasing CDD once temperatures below the CDD threshold value became frequent in mid-September, whereas K. angustifolia showed a more irregular pattern. Tissue acclimation played an important role in the decrease in A(max) as the season progressed, but only V. angustifolium showed decreasing foliar nitrogen concentrations. Based on eddy covariance flux tower data, maximum daily gross primary productivity (GPP(max)-tower) at the ecosystem level was more strongly related to CDD (r(2) = 0.59) than was maximum daily net ecosystem exchange (r(2) = 0.32). The GPP(max) was likely influenced by both tissue acclimation and the direct effects of changing temperatures and irradiances on physiological rates. Mean daily GPP, calculated for consecutive 8-day periods for a 25 km(2) area around the tower by the MODIS MOD17A2 Collection 4 satellite algorithm (GPP- MODIS), decreased more rapidly with increasing CDD than did GPP(max)-tower. Although GPP-MODIS was closely correlated with mean daily GPP from the tower (GPP(daily)-tower, r(2) = 0.95) over the late growing season, the former was about twice as high. Although MODIS estimates of air temperature closely tracked the ground data, the maximum light-use efficiency parameter used by the MODIS algorithm was much higher than that indicated by the tower measurements. There was a 3% decline in GPP(max)-tower with an increase of 10 CDD, corresponding to the percent decline in branch-level A(max) of black spruce and V. angustifolium. PMID- 15805090 TI - Contrasting net primary productivity and carbon distribution between neighboring stands of Quercus robur and Pinus sylvestris. AB - Standing biomass, net primary production (NPP) and soil carbon (C) pools were studied in a 67-year-old pedunculate oak (Quercus robur L.) stand and a neighboring 74-year- old Scots pine (Pinus sylvestris L.) stand in the Belgian Campine region. Despite a 14% lower tree density and a lower tree height in the oak stand, standing biomass was slightly higher than in the pine stand (177 and 169 Mg ha(-1) in oaks and pines, respectively), indicating that individual oak trees contained more biomass than pine trees of similar diameter. Moreover, NPP in the oak stand was more than double that in the pine stand (17.7 and 8.1 Mg ha( 1) year(-1), respectively). Several observations indicated that soil organic matter accumulated at higher rates under pines than under oaks. We therefore hypothesized that the pines were exhibiting an age-related decline in productivity due to nutrient limitation. The poor decomposability of pine litter resulted in the observed accumulation of organic matter. The subsequent immobilization of nutrients in the organic matter, combined with the already nutrient-poor soil conditions, resulted in a decrease in total NPP over time, as well as in a substantial shift in the allocation of NPP toward fine roots. In the oak stand, litter is less recalcitrant to decay and soil acidity is less severe; hence, organic matter does not accumulate and nutrients are recycled. This probably explains why NPP was much higher in the oaks than in the pines and why only a small proportion of NPP was allocated to oak fine roots. PMID- 15805091 TI - Aboveground net primary productivity of a beech (Fagus moesiaca) forest: a case study of Naousa forest, northern Greece. AB - Based on allometric relationships and information provided in forest management plans, we determined aboveground net primary productivity (ANPP) for a 10-year period in a Mediterranean beech forest (Fagus moesiaca Cz.) extending across an elevation gradient. The ANPP ranged from 1.87 to 15.71 Mg ha(-1) year(-1), and leaf area index (L*) ranged from 2.3 to 3.6. Although small trees (diameter at breast height < 10 cm) were not sampled, it was unlikely that this accounted for the low L* because there were very few small trees on a per-hectare basis. A weak positive relationship was found between ANPP and L*, and only ANPP was negatively related to elevation. Although L* did not vary with elevation, biomass growth efficiency (ANPP/L*) declined strongly with elevation. Leaf carbon isotope composition, leaf nitrogen content per unit area and specific leaf area of leaves collected from nine trees across an elevation gradient all varied significantly with elevation and were significantly related to one other, suggesting that water limitations at higher elevations may have driven the reduced growth efficiency at the stand level. Strong winds may also have negatively affected ANPP at higher elevations by altering belowground allocation. Further research is needed to test these hypotheses and to determine the belowground dynamics of phytomass in this ecosystem. PMID- 15805092 TI - Allometry and evaluation of in situ optical LAI determination in Scots pine: a case study in Belgium. AB - We evaluated several optical methods for in situ estimation of leaf area index (LAI) in a Belgian Scots pine (Pinus sylvestris L.) stand. The results obtained were compared with LAI determined from allometric relationships established in the same stand. We found high correlations between branch cross-sectional area, diameter at breast height (DBH) and basal area as dependent variables, and leaf mass, needle area and crown projection as independent variables. We then compared LAI estimated by allometry with LAI determined by three optical methods (LAI 2000, TRAC and digital hemispherical photography) both before and after corrections for blue light scattering, clumping and non-leafy material. Estimates of stand LAI of Scots pine ranged from 1.52 for hemispherical photography to 3.57 for the allometric estimate based on DBH. There was no significant difference (alpha = 0.01) between the allometric LAI estimates and the optical LAI values corrected for blue light scattering, clumping and interception by non-leafy material. However, we observed high sensitivity of the optical LAI estimates to the various conversion factors, particularly to the clumping factor, indicating the need for caution when correcting LAI measured by optical methods. PMID- 15805093 TI - Calibration and assessment of seasonal changes in leaf area index of a tropical dry forest in different stages of succession. AB - A simple measure of the amount of foliage present in a forest is leaf area index (LAI; the amount of foliage per unit ground surface area), which can be determined by optical estimation (gap fraction method) with an instrument such as the Li-Cor LAI-2000 Plant Canopy Analyzer. However, optical instruments such as the LAI-2000 cannot directly differentiate between foliage and woody components of the canopy. Studies investigating LAI and its calibration (extracting foliar LAI from optical estimates) in tropical forests are rare. We calibrated optical estimates of LAI from the LAI-2000 with leaf litter data for a tropical dry forest. We also developed a robust method for determining LAI from leaf litter data in a tropical dry forest environment. We found that, depending on the successional stage of the canopy and the season, the LAI-2000 may underestimate LAI by 17% to over 40%. In the dry season, the instrument overestimated LAI by the contribution of the woody area index. Examination of the seasonal variation in LAI for three successional stages in a tropical dry forest indicated differences in timing of leaf fall according to successional stage and functional group (i.e., lianas and trees). We conclude that when calculating LAI from optical estimates, it is necessary to account for the differences between values obtained from optical and semi-direct techniques. In addition, to calculate LAI from litter collected in traps, specific leaf area must be calculated for each species rather than from a mean value for multiple species. PMID- 15805094 TI - Long-term trends in cellulose delta13 C and water-use efficiency of tropical Cedrela and Swietenia from Brazil. AB - Elevated CO(2) concentrations ([CO(2)]) affect plant water relations and photosynthesis, and the increase in atmospheric [CO(2)] over the past 100-200 years has been related to changes in stomatal density and the carbon isotope ratio (delta(13)C) in tree rings and leaves from herbarium specimens. Because many tropical trees do not produce annual growth rings and their wood is therefore difficult to date, no trends in delta(13)C of tropical trees have been reported. Wood from Cedrela odorata L. (tropical cedar) and Swietenia macrophylla King (bigleaf mahogany), which do produce annual rings, was collected from a primary rain forest in Aripuana, Brazil (10 degrees 09' S, 59 degrees 26' W). We measured wood cellulose delta(13)C in 10-year growth increments from 37 Cedrela trees (between 11 and 151 years old in 2001) and 16 Swietenia trees (48-126 years old). A comparison of delta(13)C in cellulose of trees from different decades and of trees of different cambial ages showed that the amount of delta(13)C was largely related to the decade the wood was produced in, and not, or only to a minor extent, to tree age. Cellulose delta(13)C decreased from -26.0 to -27.3 per thousand in Cedrela and from -25.7 to -27.1 per thousand in Swietenia, with the largest changes occurring during the past 50 years. Based on these data and the trends in atmospheric [CO(2)] and delta(13)CO(2), we calculated that the internal [CO(2)] increased from about 220 to 260 ppm and that intrinsic water-use efficiency increased by 34% in Cedrela and by 52% in Swietenia. This may have implications for the water cycle and may explain the trend toward increased tree growth and turnover observed in some tropical forests. PMID- 15805095 TI - Fruit load and branch ring-barking affect carbon allocation and photosynthesis of leaf and fruit of Coffea arabica in the field. AB - Increasing fruit load (from no berries present to 25, 50 and 100% of the initial fruit load) significantly decreased branch growth on 5-year-old coffee (Coffea arabica L.) trees of the dwarf cultivar 'Costa Rica 95', during their third production cycle. Ring-barking the branches further reduced their growth. Berry dry mass at harvest was significantly reduced by increasing fruit load. Dry matter allocation to berries was four times that allocated to branch growth during the cycle. Branch dieback and berry drop were significantly higher at greater fruit loads. This illustrates the importance of berry sink strength and indicates that there is competition for carbohydrates between berries and shoots and also among berries. Leaf net photosynthesis (P(n)) increased with increasing fruit load. Furthermore, leaves of non-isolated branches bearing full fruit load achieved three times higher P(n) than leaves of isolated (ring-barked) branches without berries, indicating strong relief of leaf P(n) inhibition by carbohydrate demand from berries and other parts of the coffee tree when excess photoassimilates could be exported. Leaf P(n) was significantly higher in the morning than later during the day. This reduction in leaf P(n) is generally attributed to stomatal closure in response to high irradiance, temperature and vapor pressure deficit in the middle of the day; however, it could also be a feedback effect of reserves accumulating during the morning when climatic conditions for leaf P(n) were optimal, because increased leaf mass ratio was observed in leaves of ring-barked branches with low or no fruit loads. Rates of CO(2) emission by berries decreased and calculated photosynthetic rates of berries increased with increasing photosynthetic photon flux (PPF) especially at low PPFs (0 to 100 micromol m(-2) s(-1)). The photosynthetic contribution of berries at the bean-filling stage was estimated to be about 30% of their daily respiration costs and 12% of their total carbon requirements at PPF values commonly experienced in the field (200 to 500 micromol m(-2) s(-1)). PMID- 15805096 TI - Relationships between nutrient composition of flowers and fruit quality in orange trees grown in calcareous soil. AB - To determine if flower nutrient composition can be used to predict fruit quality, a field experiment was conducted over three seasons (1996-1999) in a commercial orange orchard (Citrus sinensis (L.) Osbeck cv. 'Valencia Late', budded on Troyer citrange rootstock) established on a calcareous soil in southern Portugal. Flowers were collected from 20 trees during full bloom in April and their nutrient composition determined, and fruits were harvested the following March and their quality evaluated. Patterns of covariation in flower nutrient concentrations and in fruit quality variables were evaluated by principal component analysis. Regression models relating fruit quality variables to flower nutrient composition were developed by stepwise selection procedures. The predictive power of the regression models was evaluated with an independent data set. Nutrient composition of flowers at full bloom could be used to predict the fruit quality variables fresh fruit mass and maturation index in the following year. Magnesium, Ca and Zn concentrations measured in flowers were related to fruit fresh mass estimations and N, P, Mg and Fe concentrations were related to fruit maturation index. We also established reference values for the nutrient composition of flowers based on measurements made in trees that produced large (> 76 mm in diameter) fruit. PMID- 15805097 TI - Anteroposterior somatotopy of innocuous cooling activation focus in human dorsal posterior insular cortex. AB - Prior data indicate that graded activation by innocuous thermal stimuli occurs in the dorsal posterior insular (dpIns) cortex of humans, rather than the parietal somatosensory regions traditionally thought necessary for discriminative somatic sensations. We hypothesized that if the dpIns subserves the haptic capacity of localization in addition to discrimination, then it should be somatotopically organized. Using functional magnetic resonance imaging to detect activation in the dpIns by graded cooling stimuli applied to the hand and neck, we found unimodal foci arranged in an anteroposterior somatotopographic pattern, consistent with participation of the dpIns in localization as well as discrimination. This gradient is orthogonal to the mediolateral somatotopy of parietal somatosensory regions, which supports the fundamental conceptual differentiation of the interoceptive somatic representation in the dpIns from the parietal exteroceptive representations. These data also support the suggestion that the poststroke central pain syndrome associated with lesions of the dpIns is a thermoregulatory dysfunction. Finally, another focus of strongly graded activation, which we interpret to represent thermoregulatory behavioral motivation elicited by dynamic cooling, was observed in the dorsal medial cortex. PMID- 15805100 TI - Hemin induces active chloride secretion in Caco-2 cells. AB - Enterocytes maintain fluid-electrolyte homeostasis by keeping a tight barrier and regulating ion channels. Carbon monoxide (CO), a product of heme degradation, modulates electrolyte transport in kidney and lung epithelium, but its role in regulating intestinal fluid-electrolyte homeostasis has not been studied. The major source of endogenous CO formation comes from the degradation of heme via heme oxygenase. We hypothesized that heme activates electrolyte transport in intestinal epithelial cells. Basolateral hemin treatment increased baseline Caco 2 cell short-circuit currents (I(sc)) twofold (control = 1.96 +/- 0.14 microA/cm(2) vs. hemin = 4.07 +/- 0.16 microA/cm(2), P < 0.01); apical hemin had no effect. Hemin-induced I(sc) was caused by Cl- secretion because it was inhibited in Cl- -free medium, with ouabain, 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), or DIDS. Apical electrogenic Na+ channel inhibitor benzamil had no effect on hemin-induced I(sc). Hemin did not alter the ability of Caco-2 cells to respond maximally to forskolin, but a soluble guanylate cyclase inhibitor, [1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ) inhibited the effects of hemin. A CO-releasing molecule, tricarbonyldichlororuthenium II, induced active Cl- secretion that was also inhibited with ODQ. We conclude that hemin induces active Cl- secretion in Caco-2 cells via a cGMP-dependent pathway. These effects are probably the consequence of CO formation. Heme and CO may be important regulators of intestinal fluid-electrolyte homeostasis. PMID- 15805101 TI - Immunolocalization and expression of kinin B1R and B2R receptors in human inflammatory bowel disease. AB - Bradykinin is a mediator of inflammation, responsible for pain, vasodilation, and capillary permeability. Bradykinin receptor 1 (B(1)R) and bradykinin receptor 2 (B(2)R) are G protein-coupled receptors that mediate kinin effects. The latter is constitutive and rapidly desensitized; the former is induced by inflammatory cytokines and resistant to densensitization. The distribution of bradykinin receptors in human intestinal tissue was studied in patients with inflammatory bowel disease (IBD), namely ulcerative colitis (UC) and Crohn's disease (CD). Both B(2)R and B(1)R proteins are expressed in the epithelial cells of normal and IBD intestines. B(1)R protein is visualized in macrophages at the center of granulomas in CD. B(2)R protein is normally present in the apexes of enterocytes in the basal area and intracellularly in inflammatory tissue. In contrast, B(1)R protein is found in the basal area of enterocytes in normal intestine but in the apical portion of enterocytes in inflamed tissue. B(1)R protein is significantly increased in both active UC and CD intestines compared with controls. In patients with active UC, B(1)R mRNA is significantly higher than B(2)R mRNA. However, in inactive UC patients, the B(1)R and B(2)R mRNA did not differ significantly. Thus bradykinin receptors in IBD may reflect intestinal inflammation. Increased B(1)R gene and protein expression in active IBD provides a structural basis of the important role of bradykinin in chronic inflammation. PMID- 15805102 TI - CD8+ cytotoxic T lymphocyte activation by soluble major histocompatibility complex-peptide dimers. AB - CD8+ cytotoxic T lymphocyte (CTL) can recognize and kill target cells that express only a few cognate major histocompatibility complex class I-peptide (pMHC) complexes. To better understand the molecular basis of this sensitive recognition process, we studied dimeric pMHC complexes containing linkers of different lengths. Although dimers containing short (10-30-A) linkers efficiently bound to and triggered intracellular calcium mobilization and phosphorylation in cloned CTL, dimers containing long linkers (> or = 80 A) did not. Based on this and on fluorescence resonance energy transfer experiments, we describe a dimeric binding mode in which two T cell receptors engage in an anti-parallel fashion two pMHC complexes facing each other with their constant domains. This binding mode allows integration of diverse low affinity interactions, which increases the overall binding and, hence, the sensitivity of antigen recognition. In proof of this, we demonstrated that pMHC dimers containing one agonist and one null ligand efficiently activate CTL, corroborating the importance of endogenous pMHC complexes in antigen recognition. PMID- 15805103 TI - Two discrete promoters regulate the alternatively spliced human interferon regulatory factor-5 isoforms. Multiple isoforms with distinct cell type-specific expression, localization, regulation, and function. AB - Interferon regulatory factor-5 (IRF-5) is a mediator of virus-induced immune activation and type I interferon (IFN) gene regulation. In human primary plasmacytoid dendritic cells (PDC), IRF-5 is transcribed into four distinct alternatively spliced isoforms (V1, V2, V3, and V4), whereas in human primary peripheral blood mononuclear cells two additional new isoforms (V5 and V6) were identified. The IRF-5 V1, V2, and V3 transcripts have different noncoding first exons and distinct insertion/deletion patterns in exon 6. Here we showed that V1 and V3 have distinct transcription start sites and are regulated by two discrete promoters. The V1 promoter (P-V1) is constitutively active, contains an IRF-E consensus-binding site, and is further stimulated in virus-infected cells by IRF family members. In contrast, endogenous V3 transcripts were up-regulated by type I IFNs, and the V3 promoter (P-V3) contains an IFN-stimulated responsive element binding site that confers responsiveness to IFN through binding of the ISGF3 complex. In addition to V5 and V6, we have identified three more alternatively spliced IRF-5 isoforms (V7, V8, and V9); V5 and V6 were expressed in peripheral blood mononuclear cells from healthy donors and in immortalized B and T cell malignancies, whereas expression of V7, V8, and V9 transcripts were detected only in human cancers. The results of this study demonstrated the existence of multiple IRF-5 spliced isoforms with distinct cell type-specific expression, cellular localization, differential regulation, and dissimilar functions in virus mediated type I IFN gene induction. PMID- 15805104 TI - Compartmentalization of lipid biosynthesis in mycobacteria. AB - The plasma membrane of Mycobacterium sp. is the site of synthesis of several distinct classes of lipids that are either retained in the membrane or exported to the overlying cell envelope. Here, we provide evidence that enzymes involved in the biosynthesis of two major lipid classes, the phosphatidylinositol mannosides (PIMs) and aminophospholipids, are compartmentalized within the plasma membrane. Enzymes involved in the synthesis of early PIM intermediates were localized to a membrane subdomain termed PMf, that was clearly resolved from the cell wall by isopyknic density centrifugation and amplified in rapidly dividing Mycobacterium smegmatis. In contrast, the major pool of apolar PIMs and enzymes involved in polar PIM biosynthesis were localized to a denser fraction that contained both plasma membrane and cell wall markers (PM-CW). Based on the resistance of the PIMs to solvent extraction in live but not lysed cells, we propose that polar PIM biosynthesis occurs in the plasma membrane rather than the cell wall component of the PM-CW. Enzymes involved in phosphatidylethanolamine biosynthesis also displayed a highly polarized distribution between the PMf and PM-CW fractions. The PMf was greatly reduced in non-dividing cells, concomitant with a reduction in the synthesis and steady-state levels of PIMs and amino phospholipids and the redistribution of PMf marker enzymes to non-PM-CW fractions. The formation of the PMf and recruitment of enzymes to this domain may thus play a role in regulating growth-specific changes in the biosynthesis of membrane and cell wall lipids. PMID- 15805105 TI - Beta ig-h3 induces keratinocyte differentiation via modulation of involucrin and transglutaminase expression through the integrin alpha3beta1 and the phosphatidylinositol 3-kinase/Akt signaling pathway. AB - Beta ig-h3 is an extracellular matrix protein whose expression is highly induced by transforming growth factor (TGF)-beta1. Whereas beta ig-h3 is known to mediate keratinocyte adhesion and migration, its effects on keratinocyte differentiation remain unclear. In the present study, it was demonstrated that expression of both beta ig-h3 and TGF-beta1 was enhanced during keratinocyte differentiation and that expression of the former was strongly induced by that of the latter. This study also asked whether changes in beta-h3 expression would affect keratinocyte differentiation. Indeed, down-regulation of beta ig-h3 by transfection with antisense beta ig-h3 cDNA constructs effectively inhibited keratinocyte differentiation by decreasing the promoter activities and thus expression of involucrin and transglutaminase. The result was an approximately 2-fold increase in mitotic capacity of the cells. Conversely, overexpression of beta ig-h3, either by transfection with beta ig-h3 expression plasmids or by exposure to recombinant beta ig-h3, enhanced keratinocyte differentiation by inhibiting cell proliferation and concomitantly increasing involucrin and transglutaminase expression. Recombinant beta ig-h3 also promoted keratinocyte adhesion through interaction with integrin alpha3beta1. Changes in beta ig-h3 expression did not affect intracellular calcium levels. Subsequent analysis revealed not only induction of Akt phosphorylation by recombinant beta ig-h3 but also blockage of Akt phosphorylation by LY294002, an inhibitor of phosphatidylinositol 3-kinase. Taken together, these findings indicate that enhanced beta ig-h3, induced by enhanced TGF-beta during keratinocyte differentiation, provoked cell differentiation by enhancing involucrin and transglutaminase expression through the integrin alpha3beta1 and phosphatidylinositol 3-kinase/Akt signaling pathway. Lastly, it was observed that beta ig-h3-mediated keratinocyte differentiation was caused by promotion of cell adhesion and not by calcium regulation. PMID- 15805106 TI - Identical Hik-Rre systems are involved in perception and transduction of salt signals and hyperosmotic signals but regulate the expression of individual genes to different extents in synechocystis. AB - In previous studies, we characterized five histidine kinases (Hiks) and the cognate response regulators (Rres) that control the expression of approximately 70% of the hyperosmotic stress-inducible genes in the cyanobacterium Synechocystis sp. PCC 6803. In the present study, we screened a gene knock-out library of Rres by RNA slot-blot hybridization and with a genome-wide DNA microarray and identified three Hik-Rre systems, namely, Hik33-Rre31, Hik10-Rre3, and Hik16-Hik41-Rre17, as well as another system that included Rre1, that were involved in perception of salt stress and transduction of the signal. We found that these Hik-Rre systems were identical to those that were involved in perception and transduction of the hyperosmotic stress signal. We compared the induction factors of the salt stress- and hyperosmotic stress-inducible genes that are located downstream of each system and found that these genes responded to the two kinds of stress to different respective extents. In addition, the Hik33-Rre31 system regulated the expression of genes that were specifically induced by hyperosmotic stress, whereas the system that included Rre1 regulated the expression of one or two genes that were specifically induced either by salt stress or by hyperosmotic stress. Our observations suggest that the perception of salt and hyperosmotic stress by the Hik-Rre systems is complex and that salt stress and hyperosmotic stress are perceived as distinct signals by the Hik-Rre systems. PMID- 15805107 TI - Phloem-localized, proton-coupled sucrose carrier ZmSUT1 mediates sucrose efflux under the control of the sucrose gradient and the proton motive force. AB - The phloem network is as essential for plants as the vascular system is for humans. This network, assembled by nucleus- and vacuole-free interconnected living cells, represents a long distance transport pathway for nutrients and information. According to the Munch hypothesis, osmolytes such as sucrose generate the hydrostatic pressure that drives nutrient and water flow between the source and the sink phloem (Munch, E. (1930) Die Stoffbewegungen in der Pflanze, Gustav Fischer, Jena, Germany). Although proton-coupled sucrose carriers have been localized to the sieve tube and the companion cell plasma membrane of both source and sink tissues, knowledge of the molecular representatives and the mechanism of the sucrose phloem efflux is still scant. We expressed ZmSUT1, a maize sucrose/proton symporter, in Xenopus oocytes and studied the transport characteristics of the carrier by electrophysiological methods. Using the patch clamp techniques in the giant inside-out patch mode, we altered the chemical and electrochemical gradient across the sucrose carrier and analyzed the currents generated by the proton flux. Thereby we could show that ZmSUT1 is capable of mediating both the sucrose uptake into the phloem in mature leaves (source) as well as the desorption of sugar from the phloem vessels into heterotrophic tissues (sink). As predicted from a perfect molecular machine, the ZmSUT1 mediated sucrose-coupled proton current was reversible and depended on the direction of the sucrose and pH gradient as well as the membrane potential across the transporter. PMID- 15805108 TI - Receptor activity-modifying protein (RAMP) isoform-specific regulation of adrenomedullin receptor trafficking by NHERF-1. AB - Receptor activity-modifying proteins (RAMPs 1-3) are single transmembrane accessory proteins critical to various G-protein coupled receptors for plasma membrane expression and receptor phenotype. A functional receptor for the vasodilatory ligand, adrenomedullin (AM), is comprised of RAMP2 or RAMP3 and calcitonin receptor-like receptor (CRLR). It is now known that RAMP3 protein protein interactions regulate the recycling of the AM2 receptor. The major aim of this study was to identify other interaction partners of RAMP3 and determine their role in CRLR-RAMP3 trafficking. Trafficking of G-protein-coupled receptors has been shown to be regulated by the Na+/H+ exchanger regulatory factor-1 (NHERF 1), an adaptor protein containing two tandem PSD-95/Discs-large/ZO-1 homology (PDZ) domains. In HEK 293T cells expressing the AM2 receptor, the complex undergoes agonist-induced desensitization and internalization. However, in the presence of NHERF-1, although the AM receptor (CRLR/RAMP3) undergoes desensitization, the internalization of the receptor complex is blocked. Overlay assays and mutational analysis indicated that RAMP3 and NHERF-1 interact via a PDZ type I domain on NHERF-1. The internalization of the CRLR-RAMP complex was not affected by NHERF-1 when CRLR was co-expressed with RAMP1 or RAMP2. Mutation of the ezrin/radixin/moesin (ERM) domain on NHERF-1 indicated that NHERF-1 inhibits CRLR/RAMP3 complex internalization by tethering the complex to the actin cytoskeleton. When examined in a primary culture of human proximal tubule cells endogenously expressing the CRLR-RAMP3 complex and NHERF-1, the CRLR-RAMP complex desensitizes but is unable to internalize upon agonist stimulation. Knock-down of either RAMP3 or NHERF-1 by RNA interference technology enabled agonist-induced internalization of the CRLR-RAMP complex. These results, using both endogenous and overexpressed cellular models, indicate a novel function for NHERF-1 and RAMP3 in the internalization of the AM receptor and suggest additional regulatory mechanisms for receptor trafficking. PMID- 15805109 TI - Structure and mode of action of the membrane-permeabilizing antimicrobial peptide pheromone plantaricin A. AB - The three-dimensional structure in dodecyl phosphocholine micelles of the 26-mer membrane-permeabilizing bacteriocin-like pheromone plantaricin A (PlnA) has been determined by use of nuclear magnetic resonance spectroscopy. The peptide was unstructured in water but became partly structured upon exposure to micelles. An amphiphilic alpha-helix stretching from residue 12 to 21 (possibly also including residues 22 and 23) was then formed in the C-terminal part of the peptide, whereas the N-terminal part remained largely unstructured. PlnA exerted its membrane-permeabilizing antimicrobial activity through a nonchiral interaction with the target cell membrane because the d-enantiomeric form had the same activity as the natural l-form. This nonchiral interaction involved the amphiphilic alpha-helical region in the C-terminal half of PlnA because a 17-mer fragment that contains the amphiphilic alpha-helical part of the peptide had antimicrobial potency that was similar to that of the l- and d-enantiomeric forms of PlnA. Also the pheromone activity of PlnA depended on this nonchiral interaction because both the l- and d-enantiomeric forms of the 17-mer fragment inhibited the pheromone activity. The pheromone activity also involved, however, a chiral interaction between the N-terminal part of PlnA and its receptor because high concentrations of the l-form (but not the d-form) of a 5-mer fragment derived from the N-terminal part of PlnA had pheromone activity. The results thus reveal a novel mechanism whereby peptide pheromones such as PlnA may function. An initial nonchiral interaction with membrane lipids induces alpha-helical structuring in a segment of the peptide pheromone. The peptide becomes thereby sufficiently structured and properly positioned in the membrane interface, thus enabling it to engage in a chiral interaction with its receptor in or near the membrane water interface. This membrane-interacting mode of action explains why some peptide pheromones/hormones such as PlnA sometimes display antimicrobial activity in addition to their pheromone activity. PMID- 15805110 TI - Cloning and functional study of porcine parotid hormone, a novel proline-rich protein. AB - A parotid gland hormone that stimulates intradentinal fluid movement is believed to play a significant role in maintaining the vitality of dentin. This hormone has been purified from porcine parotid glands and partially sequenced in our previous study (Tieche, J. M., Leonora, J., and Steinman, R. R. (1980) Endocrinology 106, 1994-2005). We now report the cloning and functional study of porcine cDNAs that code for this hormone and its complete amino acid sequence. Three cDNA clones were isolated from a porcine parotid cDNA library. The last 30 amino acids encoded by two of the cDNAs agreed with the amino acid sequence of the isolated parotid hormone. In situ hybridization and immunohistochemical staining demonstrated that the acinar cells of the parotid glands were the primary location for both the parotid hormone-related mRNAs and the translation products. A 216-bp fragment of the cDNA that contains the coding sequence for the porcine hormone was subcloned into an expression vector, and the protein expression was detected by immunoblot analysis and quantified by enzyme-linked immunosorbent assay. In addition, the 30-amino acid parotid hormone was synthesized. Both the expressed and the synthetic proteins were biologically active in that they enhanced intradentinal fluid movement as measured by intradentinal dye penetration. PMID- 15805111 TI - GABA induces activity dependent delayed-onset uncoupling of GABA/benzodiazepine site interactions in neocortical neurons. AB - Changes in the function of type A gamma-aminobutyric acid receptors (GABA(A)Rs) are associated with neuronal development and tolerance to the sedative-hypnotic effects of GABA(A)R positive modulators. Persistent activation of GABA(A)Rs by millimolar concentrations of GABA occurs under physiological conditions as GABAergic fast-spiking neurons in neocortex and cerebellum exhibit basal firing rates of 5 to 50 Hz and intermittent rates up to 250 Hz, leaving a substantial fraction of synaptic receptors occupied persistently by GABA. Persistent exposure of neurons to GABA has been shown to cause a down-regulation of receptor number and an uncoupling of GABA/benzodiazepine (BZD) site interactions with a half-life of approximately 24 h. Here, we report that a single brief exposure of neocortical neurons in primary culture to GABA for 5-10 min (t(1/2) = 3.2 +/- 0.2 min) initiates a process that results in uncoupling hours later (t(1/2) = 12.1 +/ 2.2 h). Initiation of delayed-onset uncoupling is blocked by co-incubation with picrotoxin or alpha-amanitin but is insensitive to nifedipine, indicating that uncoupling is contingent upon receptor activation and transcription but is not dependent on voltage-gated Ca2+ influx. Delayed-onset uncoupling occurs without a change in receptor number or a change in the proportion of alpha1 subunit pharmacology, as zolpidem binding affinity is unaltered. Such activity dependent latent modulation of GABA(A)R function that manifests as delayed-onset uncoupling may be relevant to physiological, pathophysiological, and pharmacological conditions where synaptic receptors are transiently exposed to GABA agonists for several minutes. PMID- 15805112 TI - Spatiotemporal switch from DeltaNp73 to TAp73 isoforms during nephrogenesis: impact on differentiation gene expression. AB - p73 is a member of the p53 gene family, which also includes p53 and p63. These proteins share sequence similarity and target genes but also have divergent roles in cancer and development. Unlike p53, transcription of the p73 gene yields multiple full-length (transactivation (TA) domain) and amino terminus-truncated (DeltaN) isoforms. DeltaNp73 acts in a dominant negative fashion to inhibit the actions of TAp73 and p53 on their target genes, promoting cell survival and proliferation and suppressing apoptosis. The balance between TAp73 and its negative regulator, DeltaNp73, may therefore represent an important determinant of developmental cell fate. There is little if anything known regarding the developmental regulation of the p73 gene. In this study, we showed that TAp73 and DeltaNp73 exhibit reciprocal spatiotemporal expression and functions during nephrogenesis. TAp73 was predominantly expressed in the differentiation domain of the renal cortex in an overlapping manner with the vasopressin-sensitive water channel aquaporin-2 (AQP-2). Chromatin immunoprecipitation assays demonstrated that the endogenous AQP-2 promoter was occupied by TAp73 in a developmentally regulated manner. Furthermore TAp73 stimulated AQP-2 promoter-driven reporter expression. TAp73 also activated the bradykinin B2 receptor (B2R) promoter, a developmentally regulated gene involved in regulation of sodium excretion. The transcriptional effects of TAp73 on AQP-2 and B2R were independent of p53. In marked contrast to TAp73, DeltaNp73 isoforms were induced early in development and were preferentially expressed in proliferating nephron precursors. Moreover DeltaNp73 was a potent repressor of B2R gene transcription. We conclude that the p73 gene is developmentally regulated during kidney organogenesis. The spatiotemporal switch from DeltaNp73 to TAp73 may play an important role in the terminal differentiation program of the developing nephron. PMID- 15805113 TI - Down-regulation of BRCA2 expression by collagen type I promotes prostate cancer cell proliferation. AB - BRCA2 is a tumor suppressor gene that when mutated confers an increased susceptibility to developing breast and prostate carcinoma. Besides its role in mediating DNA repair, new evidence suggests that BRCA2 may also play a role in suppressing cancer cell growth. Because altered interactions between neoplastic cells and the surrounding extracellular matrix (ECM) play a pivotal role in unchecked cancer cell proliferation and metastatic progression, we hypothesized that the ECM may have an effect in BRCA2 expression. By using normal and prostate carcinoma cell lines, we demonstrated that although normal cells transiently increase BRCA2 protein levels when adhering to the ECM protein collagen type I (COL1), carcinoma cells exhibit a significant reduction in BRCA2 protein. This aberrant effect is independent from de novo protein synthesis and results from COL1-beta(1) integrin signaling through phosphatidylinositol (PI) 3-kinase leading to BRCA2 ubiquitination and degradation in the proteasome. BRCA2 protein depletion after cancer cell adhesion to COL1 or in small RNA interference assays triggers new DNA synthesis, a trophic effect that is abrogated by recombinant BRCA2 expression. Blocking or inhibiting beta(1) integrin, PI 3-kinase, or proteasome activity all have a negative effect on COL1-mediated DNA synthesis in cancer cells. In normal cells, the transient increase in BRCA2 expression is independent from beta(1) integrin or PI 3-kinase and has no effect in cell proliferation. In summary, these results unravel a novel mechanism whereby prostate carcinoma cell proliferation is enhanced by the down-regulation of BRCA2 expression when interacting with COL1, a major component of the ECM at osseous metastatic sites. PMID- 15805114 TI - Differential trafficking of GluR7 kainate receptor subunit splice variants. AB - Kainate receptors (KARs) are heteromeric ionotropic glutamate receptors that play a variety of roles in the regulation of synaptic network activity. The function of glutamate receptors (GluRs) is highly dependent on their surface density in specific neuronal domains. Alternative splicing is known to regulate surface expression of GluR5 and GluR6 subunits. The KAR subunit GluR7 exists under different splice variant isoforms in the C-terminal domain (GluR7a and GluR7b). Here we have studied the trafficking of GluR7 splice variants in cultured hippocampal neurons from wild-type and KAR mutant mice. We have found that alternative splicing regulates surface expression of GluR7-containing KARs. GluR7a and GluR7b differentially traffic from the ER to the plasma membrane. GluR7a is highly expressed at the plasma membrane, and its trafficking is dependent on a stretch of positively charged amino acids also found in GluR6a. In contrast, GluR7b is detected at the plasma membrane at a low level and retained mostly in the endoplasmic reticulum (ER). The RXR motif of GluR7b does not act as an ER retention motif, at variance with other receptors and ion channels, but might be involved during the assembly process. Like GluR6a, GluR7a promotes surface expression of ER-retained subunit splice variants when assembled in heteromeric KARs. However, our results also suggest that this positive regulation of KAR trafficking is limited by the ability of different combinations of subunits to form heteromeric receptor assemblies. These data further define the complex rules that govern membrane delivery and subcellular distribution of KARs. PMID- 15805115 TI - Phycobilisome linker proteins are phosphorylated in Synechocystis sp. PCC 6803. AB - The controversial issue of protein phosphorylation from the photosynthetic apparatus of Synechocystis sp. PCC 6803 has been reinvestigated using new detection tools that include various immunological and in vivo labeling approaches. The set of phosphoproteins detected with these methods includes ferredoxin-NADPH reductase and the linker proteins of the phycobilisome antenna. Using mutants that lack a specific set of linker proteins and are affected in phycobilisome assembly, we show that the phosphoproteins from the phycobilisomes correspond to the membrane, rod, and rod-core linkers. These proteins are in a phosphorylated state within the assembled phycobilisomes. Their dephosphorylation requires partial disassembly of the phycobilisomes and further contributes to their complete disassembly in vitro. In vivo we observed linker dephosphorylation upon long-term exposure to higher light intensities and under nitrogen limitation, two conditions that lead to remodeling and turnover of phycobilisomes. We conclude that this phosphorylation process is instrumental in the regulation of assembly/disassembly of phycobilisomes and should participate in signaling for their proteolytic cleavage and degradation. PMID- 15805117 TI - Proliferating cell nuclear antigen (PCNA) may function as a double homotrimer complex in the mammalian cell. AB - The diverse function of proliferating cell nuclear antigen (PCNA) may be regulated by interactions with different protein partners. Interestingly, the binding sites for all known PCNA-associating proteins are on the outer surface or the C termini ("front") sides of the PCNA trimer. Using cell extracts and purified human PCNA protein, we show here that two PCNA homotrimers form a back to-back doublet. Mutation analysis suggests that the Arg-5 and Lys-110 residues on the PCNA back side are the contact points of the two homotrimers in the doublet. Furthermore, short synthetic peptides encompassing either Arg-5 or Lys 110 inhibit double trimer formation. We also found that a PCNA double trimer, but not a homotrimer alone, can simultaneously accommodate chromatin assembly factor 1 and polymerase delta. Together, our data supports a model that chromatin remodeling by chromatin assembly factor-1 (and, possibly, many other cellular activities) are tightly coupled with DNA replication (and repair) through a PCNA double trimer complex. PMID- 15805118 TI - Inducible nitric-oxide synthase and NO donor induce insulin receptor substrate-1 degradation in skeletal muscle cells. AB - Chronic inflammation plays an important role in insulin resistance. Inducible nitric-oxide synthase (iNOS), a mediator of inflammation, has been implicated in many human diseases including insulin resistance. However, the molecular mechanisms by which iNOS mediates insulin resistance remain largely unknown. Here we demonstrate that exposure to NO donor or iNOS transfection reduced insulin receptor substrate (IRS)-1 protein expression without altering the mRNA level in cultured skeletal muscle cells. NO donor increased IRS-1 ubiquitination, and proteasome inhibitors blocked NO donor-induced reduction in IRS-1 expression in cultured skeletal muscle cells. The effect of NO donor on IRS-1 expression was cGMP-independent and accentuated by concomitant oxidative stress, suggesting an involvement of nitrosative stress. Inhibitors for phosphatidylinositol-3 kinase, mammalian target of rapamycin, and c-Jun amino-terminal kinase failed to block NO donor-induced IRS-1 reduction, whereas these inhibitors prevented insulin stimulated IRS-1 decrease. Moreover iNOS expression was increased in skeletal muscle of diabetic (ob/ob) mice compared with lean wild-type mice. iNOS gene disruption or treatment with iNOS inhibitor ameliorated depressed IRS-1 expression in skeletal muscle of diabetic (ob/ob) mice. These findings indicate that iNOS reduces IRS-1 expression in skeletal muscle via proteasome-mediated degradation and thereby may contribute to obesity-related insulin resistance. PMID- 15805119 TI - c-myc Promoter-binding protein 1 (MBP-1) regulates prostate cancer cell growth by inhibiting MAPK pathway. AB - Prostate cancer is the most common and invasive type of cancer among American men, and the second leading cause of cancer-elated deaths in the United States. Unfortunately, an effective therapeutic regimen is still lacking for advance stages of the disease. Recently, MEK5 has been shown to overexpress in prostate cancer and is associated with poor survival outcome. MEK5 exists as alpha- and beta-isoforms. MEK5alpha induces cell proliferation by activating its downstream molecules, whereas MEK5beta expression is associated with inhibition of cell growth. We have recently shown that exogenous expression of c-myc promoter binding protein 1 (MBP-1) induces prostate cancer cell death (Ghosh, A. K., Steele, R., and Ray, R. B. (2005) Cancer Res. 65, 718-721). In this study, we have investigated whether inhibition of MEK5 signaling pathway can modulate prostate cancer cell growth. MBP-1 is a general transcriptional repressor and modulates a number of cellular genes. Therefore, we examined the endogenous expression status of MEK5 in androgen-independent prostate cancer cells upon recombinant adenovirus-mediated introduction of MBP-1. Our results demonstrated that MBP-1 expression reduced the endogenous MEK5alpha protein level; on the other hand, MEK5beta expression was enhanced significantly. Transduction of MBP-1 modulates the downstream signaling molecules of MEK5, such as activation of the cyclin D1 promoter and MEF2C transcriptional activities in androgen-independent prostate cancer cells. MBP-1 expression also modulates MEK5-mediated activation of NF-kappaB. Further analysis suggested that MBP-1 physically associates with MEK5 and induces proteasome-mediated degradation of the MEK5 protein, which appears to occur independently of ubiquitination. Together, our results suggested a novel role of MBP-1 for suppression of prostate cancer cell growth by regulating the MEK5-mediated signaling pathway. PMID- 15805120 TI - Determination of tumor necrosis factor receptor-associated factor trimerization in living cells by CFP->YFP->mRFP FRET detected by flow cytometry. AB - The availability of protein fluorophores with appropriate spectral properties has made it possible to employ fluorescence resonance energy transfer (FRET) to assess interactions between three proteins microscopically. Flow cytometry offers excellent sensitivity, effective signal separation and the capacity to assess a large number of events, and, therefore, should be an ideal means to explore protein interactions in living cells. Here, we report a flow-cytometric FRET technique that employed both direct energy transfer from CFP-->YFP-->mRFP and donor quenching to assess TRAF2 trimerization in living cells. Initially, a series of fusion proteins incorporating CFP, YFP and mRFP with spacers that did or did not permit FRET were employed to document the magnitude of CFP-->YFP and YFP-->mRFP FRET and to calculate the efficiency of CFP-->YFP-->mRFP two-step FRET. Based upon this, TRAF2 homotrimerization could be detected. This method should have great utility in studying the dynamics of interactions between three specific proteins in vivo. PMID- 15805121 TI - An RNA polymerase II construct synthesizes short-hairpin RNA with a quantitative indicator and mediates highly efficient RNAi. AB - RNA interference (RNAi) mediates gene silencing in many eukaryotes and has been widely used to investigate gene functions. A common method to induce sustained RNAi is introducing plasmids that synthesize short hairpin RNAs (shRNAs) using Pol III promoters. While these promoters synthesize shRNAs and elicit RNAi efficiently, they lack cell specificity. Monitoring shRNA expression levels in individual cells by Pol III promoters is also difficult. An alternative way to deliver RNAi is to use Pol II-directed synthesis of shRNA. Previous efforts in developing a Pol II system have been sparse and the results were conflicting, and the usefulness of those Pol II vectors has been limited due to low efficacy. Here we demonstrate a new Pol II system that directs efficient shRNA synthesis and mediates strong RNAi at levels that are comparable with the commonly used Pol III systems. In addition, this system synthesizes a marker protein under control of the same promoter as the shRNA, thus providing an unequivocal indicator, not only to the cells that express the shRNA, but also to the levels of the shRNA expression. This system may be adapted for in vivo shRNA expression and gene silencing. PMID- 15805122 TI - Practical lessons from protein structure prediction. AB - Despite recent efforts to develop automated protein structure determination protocols, structural genomics projects are slow in generating fold assignments for complete proteomes, and spatial structures remain unknown for many protein families. Alternative cheap and fast methods to assign folds using prediction algorithms continue to provide valuable structural information for many proteins. The development of high-quality prediction methods has been boosted in the last years by objective community-wide assessment experiments. This paper gives an overview of the currently available practical approaches to protein structure prediction capable of generating accurate fold assignment. Recent advances in assessment of the prediction quality are also discussed. PMID- 15805123 TI - Structure of HinP1I endonuclease reveals a striking similarity to the monomeric restriction enzyme MspI. AB - HinP1I, a type II restriction endonuclease, recognizes and cleaves a palindromic tetranucleotide sequence (G/CGC) in double-stranded DNA, producing 2 nt 5' overhanging ends. Here, we report the structure of HinP1I crystallized as one protein monomer in the crystallographic asymmetric unit. HinP1I displays an elongated shape, with a conserved catalytic core domain containing an active-site motif of SDX18QXK and a putative DNA-binding domain. Without significant sequence homology, HinP1I displays striking structural similarity to MspI, an endonuclease that cleaves a similar palindromic DNA sequence (C/CGG) and binds to that sequence crystallographically as a monomer. Almost all the structural elements of MspI can be matched in HinP1I, including both the DNA recognition and catalytic elements. Examining the protein-protein interactions in the crystal lattice, HinP1I could be dimerized through two helices located on the opposite side of the protein to the active site, generating a molecule with two active sites and two DNA-binding surfaces opposite one another on the outer surfaces of the dimer. A possible functional link between this unusual dimerization mode and the tetrameric restriction enzymes is discussed. PMID- 15805124 TI - Acetylation of a fucosyl residue at the reducing end of Mesorhizobium loti nod factors is not essential for nodulation of Lotus japonicus. AB - NodMl-V(C(18:1), Me, Cb, AcFuc) is a major component of lipo-chitin oligosaccharides (LCOs), or Nod factors, produced by Mesorhizobium loti. The presence of a 4-O-acetylated fucosyl residue (AcFuc) at the reducing end has been thought to be essential for symbiotic interactions with the compatible host plant, Lotus japonicus. We generated an M. loti mutant in which the nolL gene is disrupted; nolL has been shown to encode acetyltransferase that is responsible for acetylation of the fucosyl residue. The nolL disruptant Ml107 produced LCOs that lacked acetylation of fucosyl residues as expected, but exhibited nodulation performance on L. japonicus as efficiently as the wild-type M. loti strain MAFF303099. We show that LCOs without acetylation of a fucosyl residue purified from Ml107 are also able to induce abundant root hair deformation and nodule primordium formation. These results indicate that NolL-dependent acetylation of a fucosyl residue at the reducing end of M. loti LCOs is not essential for nodulation of L. japonicus. PMID- 15805126 TI - The prognosis of different fatigue diagnostic labels: a longitudinal survey. AB - BACKGROUND: Several different diagnostic labels exist for the fatigue syndromes, including chronic fatigue syndrome (CFS), myalgic encephalomyelitis (ME) and postviral fatigue syndrome (PVFS). An allied condition is fibromyalgia. No study has examined prognostic differences across these different labels. OBJECTIVE: To compare the prognoses of patients labelled with different fatigue syndromes in primary care. METHODS: We performed a longitudinal survey, using electronic records from the General Practice Research Database. All 18,122 patients diagnosed by their GP with a fatigue syndrome from 1988-2001 with a minimum of one year of records after diagnosis were collated into four groups: CFS, ME, PVFS and fibromyalgia. CFS and ME were combined for the main analysis as no code for CFS was available until 1995. The length of illness was calculated as the interval between the diagnosis and the last recorded fatigue symptom, expressed as days per year, to account for differing lengths of record after diagnosis. RESULTS: Patients with CFS/ME combined had a worse prognosis (median length of illness 80 days per year; interquartile range 0-242) than fibromyalgia (51; 0 244) or PVFS 0 (0-108), a significant difference, P < 0.001. In a subgroup analysis, ME had a worse prognosis (median length of illness in days per year 106; interquartile range 0-259) than CFS (33; 0-170), P < 0.001, in spite of a better course before diagnosis. Secondary outcome measures were consistent with these results. CONCLUSION: There were important differences in outcome between the various fatigue labels, with ME having the worst prognosis and PVFS the best. This could be an adverse effect of the label ME itself. Alternatively, patients who are destined to have a worse prognosis may preferentially attract the ME label. Our data support the first interpretation. PMID- 15805125 TI - Role of multivitamins and mineral supplements in preventing infections in elderly people: systematic review and meta-analysis of randomised controlled trials. AB - OBJECTIVE: To evaluate the effectiveness of multivitamins and mineral supplements in reducing infections in an elderly population. DESIGN: Systematic review and meta-analysis of randomised controlled trials. DATA SOURCES: Medline and other databases. Reference lists of identified articles were inspected for further relevant articles. SELECTION OF STUDIES: Trials were included if they evaluated the effect of multivitamins and mineral supplements on infections in an elderly population. REVIEW METHODS: Studies were assessed for the methodological quality by using the Jadad instrument. If the data required for the analyses were not available from the published articles we requested them from the original study authors. Meta-analysis was undertaken on three outcomes: the mean difference in number of days spent with infection, the odds ratio of at least one infection in the study period, and the incidence rate ratio for the difference in infection rates. Data on adverse events were also extracted. RESULTS: Eight trials met our inclusion criteria. Owing to inconsistency in the outcomes reported, only a proportion of the trials could be included in each meta-analysis. Multivitamins and mineral supplements were found to reduce the mean annual number of days spent with infection (three studies) by 17.5 (95% confidence interval 11 to 24, P < 0.001). The odds ratio for at least one infection in the study period (three studies) was 1.10 (0.81 to 1.50, P = 0.53). The infection rate ratio (four studies) was 0.89 (0.78 to 1.03, P = 0.11). Reporting of adverse events was poor. CONCLUSION: The evidence for routine use of multivitamin and mineral supplements to reduce infections in elderly people is weak and conflicting. Study results are heterogeneous, and this is partially confounded by outcome measure. PMID- 15805127 TI - Prognostic factors for serious morbidity and mortality from community-acquired lower respiratory tract infections among the elderly in primary care. AB - BACKGROUND: Uncertainty about the prognosis of lower respiratory tract infections (LRTI) hinders optimal management in primary care. OBJECTIVE: We determined prognostic factors for a severe complicated course of LRTI among elderly patients in primary care. METHODS: In a retrospective clinical database study we examined 455 patients with a first LRTI episode; 226 with physician-diagnosed acute bronchitis or lung exacerbations and 229 with pneumonia. Multivariate logistic regression analysis was used to assess independent associations of the potential predictors with the endpoint. RESULTS: Occurrence of the combined endpoint 30-day home-treated complications from LRTI (4.4%) or hospitalisation (4.6%), or all cause mortality (5.3%) was 14.3%. In a logistic regression model, increasing age [odds ratio (OR) 1.04; 95% confidence interval (95% CI) 1.00-1.08], male sex (OR 3.12; 95% CI 1.66-5.87), heart failure (OR 5.14; 95% CI 2.33-11.34), stroke or dementia (OR 3.36; 95% CI 1.18-9.58), use of antidepressants or benzodiazepines (OR 1.89; 95% CI 1.02-3.52) and a diagnosis of pneumonia (OR 4.24; 95% CI 2.17 8.28) were independent predictors. CONCLUSION: GPs need to be aware of readily available prognostic factors that can be used in primary care to complement physical examination and laboratory data in LRTI to guide preventive and therapeutic management decisions. PMID- 15805128 TI - Chronic Fatigue Syndrome: a survey of GPs' attitudes and knowledge. AB - BACKGROUND: GPs need evidence and guidance to help them diagnose and manage Chronic Fatigue Syndrome (CFS)/ME appropriately. OBJECTIVES: The aim of this survey was to obtain baseline data and identify the factors associated with GPs' attitudes to and knowledge of CFS/ME. The attitude of GPs to the condition is an important indicator of likely prognosis. METHODS: A postal questionnaire was sent to 1054 GPs served by Taunton, Bristol and Gloucester laboratories. GPs' attitudes to nine statements about CFS/ME were assessed and the factors associated with positive or negative responses were determined. Knowledge of the clinical features was also assessed. RESULTS: 811 GPs (77%) returned the questionnaire. 48% of GPs did not feel confident with making a diagnosis of CFS/ME and 41% did not feel confident in treatment. 72% of GPs accepted CFS/ME as a recognisable clinical entity and those GPs had significantly more positive attitudes. Three other key factors that were significantly, positively associated with GPs' attitudes were knowing someone socially with CFS/ME, being male and seeing more patients with the condition in the last year. CONCLUSION: Despite the publication of guidance for GPs on CFS/ME, confidence with making a diagnosis and management was found to be low. Educational initiatives and guidance for GPs should stress the importance of accepting CFS/ME as a recognisable clinical entity, as this is linked to having a positive attitude and could lead to improved confidence to make a diagnosis and treat CFS/ME patients. PMID- 15805129 TI - A qualitative study of choosing and using an NHS Walk-in Centre. AB - BACKGROUND: NHS Walk-in Centres have been introduced to improve access to healthcare in the UK. Little is understood about why people choose Walk-in Centres from among the range of options available to them. OBJECTIVES: To explore users' accounts of choosing and using an NHS Walk-in Centre. METHODS: Semi structured interviews with 23 users who had recently attended an NHS Walk-in Centre were conducted. Analysis was based on the constant comparative method. RESULTS: Participants' accounts revealed two types of service use: those who knew what was wrong with them and had a clear idea of what treatment was required, and those seeking professional advice. Users reported "solidarity" with the NHS and other NHS users, and were highly sensitive to the demands on both Accident and Emergency and GP services in their choice of services. The Walk-in Centre appeared to function as a means of overcoming the barriers to healthcare associated with other healthcare services, although there was some lack of clarity about the purpose of the Walk-in Centre. CONCLUSIONS: Users' accounts suggest that NHS Walk-in Centres improve access to healthcare by opening up an alternative means of seeking a professional opinion or treatment. It is especially important in allowing people to use the NHS without feeling that they are increasing the burden on general practice and A&E facilities, and to feel that they are behaving responsibly while still meeting their own needs. PMID- 15805130 TI - A randomized controlled trial of the effect of providing information on accidental injury admissions and their costs to Primary Care Groups and Trusts. AB - BACKGROUND: Primary Care Groups and Trusts (PCG/Ts) are responsible for improving the health of local populations, but there is little evidence of their strategic involvement in accident prevention. OBJECTIVE: To determine the effect of providing information on local accidental injuries to PCG/Ts on the development of accident prevention strategies. METHODS: The study is a randomized controlled trial in PCG/Ts in the former Trent Region. Intervention PCG/Ts were sent profiles containing PCG/T specific information on hospital admissions for accidental injury and their costs and on accident prevention interventions. Health promotion leads were surveyed at baseline and three and fifteen months. The primary outcome was whether the PCG/T had a written accident prevention strategy. Secondary outcome measures included other accident prevention activities, prioritization of accident prevention, and knowledge and attitudes towards accident prevention. RESULTS: At three months 38% of PCG/Ts in the intervention arm had a written accident prevention strategy, compared with 42% in the control arm [difference -4%, 95% confidence interval (CI) -31% to 25%]. At fifteen months these values were 55% in the intervention arm and 50% in the control arm (difference 5%, CI -24% to 33%). There were no statistically significant differences between the groups for any outcome measures. CONCLUSIONS: Providing profiles containing information on accidental injuries to PCG/Ts did not significantly increase the development of accident prevention strategies, however this study only had sufficient power to detect substantial changes. Larger national studies would be required to detect moderate effects of tailored packages in changing PCG/T behaviour. PMID- 15805131 TI - Improving patient-provider communication: insights from interpreters. AB - BACKGROUND: It is important for physicians to recognize and address potential cross-cultural communication barriers with their patients. Several studies have demonstrated the importance of trained medical interpreters for ensuring effective patient-provider communication. Medical interpreters also represent an untapped source of insight into common communication problems. Such insights can contribute to strengthening physicians' cross cultural communication skills. OBJECTIVE: The purpose of this study was to explore professional medical interpreters' experiences and perspectives regarding patient-provider communication difficulties. METHODS: Key informant interviews were conducted with a purposive sample of nine professional interpreters working at the Geneva University Hospitals general medicine outpatient clinic. RESULTS: Interpreters described three domains where physicians and patients were likely to differ, and where mutual lack of awareness of those differences could lead to misunderstandings. These were: (1) ideas about the patient's health problem; (2) expectations of the clinical encounter; and (3) verbal and non-verbal communication styles. Interpreters recommended that cultural competence training for physicians focus on raising awareness about potential sources of misunderstanding and about the difficulties inherent in medical translation; providing basic background knowledge of patients' countries of origin; and adapting to patients' communication styles. While physicians' own perceptions of communication difficulties are important for developing learner-centered training activities, interpreters' bilingual and bicultural position allows for the identification of communication barriers that may be difficult for physicians to recognize. PMID- 15805132 TI - The costs of care in general practice: patients compared by the council tax valuation band of their home address. AB - BACKGROUND: It is difficult to measure and compare workload in UK general practice. A GP/health economist team recently proposed a means of calculating the unit cost of a GP consulting. It is therefore now possible to extrapolate to the costs of other clinical tasks in a practice and then to compare the workloads of caring for different patients and compare between practices. OBJECTIVES: The study aims were: (i) to estimate the relative costs of daily clinical activities within a practice (implying workload); and (ii) to compare the costs of caring for different types of patients categorized by gender, by age, and by socio economic status as marked by the Council Tax Valuation Band (CTVB) of home address. METHODS: The study design was a cross-sectional cost comparison of all clinical activity aggregated, by patient, over one year in an English semi-rural general practice. The subjects were 3339 practice patients, randomly selected. The main outcome measures were costs per clinical domain and overall costs per patient per year; both then compared by gender, age group and by CTVB. RESULTS: CTVB is as significant a predictor of patient care cost (workload) as is patient gender and age (both already known). CONCLUSIONS: It is now possible to estimate the cost of care of different patients in such a way that NHS planning and especially resource allocation to practices could be improved. PMID- 15805134 TI - General practices as emergent research organizations: a qualitative study into organizational development. AB - BACKGROUND: An increasing proportion of research in primary care is locally undertaken in designated research practices. Capacity building to support high quality research at these grass roots is urgently needed and is a government priority. There is little previously published research on the process by which GP practices develop as research organizations or on their specific support needs at organizational level. METHODS: Using in-depth qualitative interviews with 28 key informants in 11 research practices across the UK, we explored their historical accounts of the development of research activity. We analysed the data with reference to contemporary theories of organizational development. RESULTS: Participants identified a number of key events and processes, which allowed us to produce a five-phase model of practice development in relation to research activity (creative energy, concrete planning, transformation/differentiation, consolidation and collaboration). Movement between these phases was not linear or continuous, but showed emergent and adaptive properties in which specific triggers and set-backs were often critical. CONCLUSION: This developmental model challenges previous categorical taxonomies of research practices. It forms a theory-driven framework for providing appropriate support at the grass roots of primary care research, based on the practice's phase of development and the nature of external triggers and potential setbacks. Our findings have important implications for the strategic development of practice-based research in the UK, and could serve as a model for the wider international community. PMID- 15805133 TI - Obtaining optimal control in mild asthma: theory and practice. AB - BACKGROUND: Studies have shown that asthma severity is easily under-estimated and as a result, patients may be under-treated with reduced asthma control. OBJECTIVE: This study, performed in the General Practice Research Database (GPRD), investigates asthma control in patients treated as intermittent asthmatics (short-acting beta agonist (SABA) alone), or persistent asthmatics (additional inhaled cortico-steroid (ICS), no other medication). METHODS: Patients (0-45 years) diagnosed with asthma between 1 January 1995 and 31 December 2001 taking > or =2 scripts for SABA (SABA only group) or > or =3 scripts for ICS (ICS group) in the first six months following diagnosis were selected. Factors associated with drug prescriptions were assessed. RESULTS: SABA script rates were 3.6 and 5.1 per year in the SABA and ICS group respectively, i.e. >1 dose/day. 10.5% of SABA group and 13.4% of ICS group used oral steroids. Within the SABA group, 37% were stepped up to ICS, the time to first ICS script being significantly associated with prior hospitalization (RR 2.26, CI 1.65-3.10) and atopy (RR 1.47, CI 1.33-1.63). A higher rate of oral steroid use was significantly associated with using ICS, being female, adult and smoking. Smokers and atopic individuals had increased risk of obtaining an earlier script for oral steroid (RR 1.32, CI 1.10-1.59 and RR 1.28, CI 1.10-1.49, respectively). CONCLUSIONS: Asthma control was sub-optimal in a substantial proportion of patients using relatively high doses of SABA, or SABA and ICS from the outset of asthma treatment in general practice. Being female, atopic, a smoker and prior hospitalization were all associated with lack of asthma control and could guide physicians in treatment prescribing. PMID- 15805135 TI - Photography as a metaphor for (epidemiological) research. PMID- 15805136 TI - E. coli virulence factor hemolysin induces neutrophil apoptosis and necrosis/lysis in vitro and necrosis/lysis and lung injury in a rat pneumonia model. AB - Enteric gram-negative bacilli, such as Escherichia coli are the most common cause of nosocomial pneumonia. In this study a wild-type extraintestinal pathogenic strain of E. coli (ExPEC)(CP9) and isogenic derivatives deficient in hemolysin (Hly) and cytotoxic necrotizing factor (CNF) were assessed in vitro and in a rat model of gram-negative pneumonia to test the hypothesis that these virulence factors induce neutrophil apoptosis and/or necrosis/lysis. As ascertained by in vitro caspase-3/7 and LDH activities and neutrophil morphology, Hly mediated neutrophil apoptosis at lower E. coli titers (1 x 10(5-6) cfu) and necrosis/lysis at higher titers (> or =1 x 10(7) cfu). Data suggest that CNF promotes apoptosis but not necrosis or lysis. We also demonstrate that annexin V/7-amino-actinomycin D staining was an unreliable assessment of apoptosis using live E. coli. The use of caspase-3/7 and LDH activities and neutrophil morphology supported the notion that necrosis, not apoptosis, was the primary mechanism by which neutrophils were affected in our in vivo gram-negative pneumonia model using live E. coli. In addition, in vivo studies demonstrated that Hly mediates lung injury. Neutrophil necrosis was not observed when animals were challenged with purified lipopolysaccharide, demonstrating the importance of using live bacteria. These findings establish that Hly contributes to ExPEC virulence by mediating neutrophil toxicity, with necrosis/lysis being the dominant effect of Hly on neutrophils in vivo and by lung injury. Whether Hly-mediated lung injury is due to neutrophil necrosis, a direct effect of Hly, or both is unclear. PMID- 15805137 TI - Nuclear localization of leukotriene A4 hydrolase in type II alveolar epithelial cells in normal and fibrotic lung. AB - Leukotriene A4 (LTA4) hydrolase catalyzes the final step in leukotriene B4 (LTB4) synthesis. In addition to its role in LTB4 synthesis, the enzyme possesses aminopeptidase activity. In this study, we sought to define the subcellular distribution of LTA4 hydrolase in alveolar epithelial cells, which lack 5 lipoxygenase and do not synthesize LTA4. Immunohistochemical staining localized LTA4 hydrolase in the nucleus of type II but not type I alveolar epithelial cells of normal mouse, human, and rat lungs. Nuclear localization of LTA4 hydrolase was also demonstrated in proliferating type II-like A549 cells. The apparent redistribution of LTA4 hydrolase from the nucleus to the cytoplasm during type II to-type I cell differentiation in vivo was recapitulated in vitro. Surprisingly, this change in localization of LTA4 hydrolase did not affect the capacity of isolated cells to convert LTA4 to LTB4. However, proliferation of A549 cells was inhibited by the aminopeptidase inhibitor bestatin. Nuclear accumulation of LTA4 hydrolase was also conspicuous in epithelial cells during alveolar repair following bleomycin-induced acute lung injury in mice, as well as in hyperplastic type II cells associated with fibrotic lung tissues from patients with idiopathic pulmonary fibrosis. These results show for the first time that LTA4 hydrolase can be accumulated in the nucleus of type II alveolar epithelial cells and that redistribution of the enzyme to the cytoplasm occurs with differentiation to the type I phenotype. Furthermore, the aminopeptidase activity of LTA4 hydrolase within the nucleus may play a role in promoting epithelial cell growth. PMID- 15805138 TI - HIV-1 Tat protein-induced VCAM-1 expression in human pulmonary artery endothelial cells and its signaling. AB - Expression of cell adhesion molecule in endothelial cells upon activation by human immunodeficiency virus (HIV) infection is associated with the development of atherosclerotic vasculopathy. We postulated that induction of vascular cell adhesion molecule-1 (VCAM-1) by HIV-1 Tat protein in endothelial cells might represent an early event that could culminate in inflammatory cell recruitment and vascular injury. We determined the role of HIV-1 Tat protein in VCAM-1 expression in human pulmonary artery endothelial cells (HPAEC). HIV-1 Tat protein treatment significantly increased cell-surface expression of VCAM-1 in HPAEC. Consistently, mRNA expression of VCAM-1 was also increased by HIV-1 Tat protein as measured by RT-PCR. HIV-1 Tat protein-induced VCAM-1 expression was abolished by the NF-kappaB inhibitor pyrrolidine dithiocarbamate (PDTC) and the p38 MAPK inhibitor SB-203580. Furthermore, HIV-1 Tat protein enhanced DNA binding activity of NF-kappaB, facilitated nuclear translocation of NF-kappaB subunit p65, and increased production of reactive oxygen species (ROS). Similarly to VCAM-1 expression, HIV-1 Tat protein-induced NF-kappaB activation and ROS generation were abrogated by PDTC and SB-203580. These data indicate that HIV-1 Tat protein is able to induce VCAM-1 expression in HPAEC, which may represent a pivotal early molecular event in HIV-induced vascular/pulmonary injury. These data also suggest that the molecular mechanism underlying the HIV-1 Tat protein-induced VCAM-1 expression may involve ROS generation, p38 MAPK activation, and NF-kappaB translocation, which are the characteristics of pulmonary endothelial cell activation. PMID- 15805139 TI - Differential response of TIMP-3 null mice to the lung insults of sepsis, mechanical ventilation, and hyperoxia. AB - An imbalance in matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs) leads to excessive or insufficient tissue breakdown, which is associated with many disease processes. The TIMP-3 null mouse is a model of MMP/TIMP imbalance, which develops air space enlargement and decreased lung function. These mice responded differently to cecal ligation and perforation (CLP)-induced septic lung injury than wild-type controls. The current study addresses whether the TIMP-3 knockout lung is susceptible to different types of insults or only those involving sepsis, by examining its response to lipopolysaccharide (LPS)-induced sepsis, mechanical ventilation (MV), and hyperoxia. TIMP-3 null noninjured controls of each insult consistently demonstrated significantly higher compliance vs. wild-type mice. Null mice treated with LPS had a further significantly increased compliance compared with untreated controls. Conversely, MV and hyperoxia did not alter compliance in the null lung. MMP abundance and activity increased in response to LPS but were generally unaltered following MV or hyperoxia, correlating with compliance alterations. All three insults produced inflammatory cytokines; however, the response of the null vs. wild-type lung was dependent on the type of insult. Overall, this study demonstrated that 1) LPS-induced sepsis produced a similar response in null mice to CLP-induced sepsis, 2) the null lung responded differently to various insults, and 3) the null susceptibility to compliance changes correlated with increased MMPs. In conclusion, this study provides insight into the role of TIMP-3 in response to various lung insults, specifically its importance in regulating MMPs to maintain compliance during a sepsis. PMID- 15805140 TI - Haemodynamic effects of haemorrhage during xenon anaesthesia in pigs. AB - BACKGROUND: It was hypothesized that xenon would stabilize mean arterial pressure (MAP) in haemorrhagic shock, recovery, and volume resuscitation, because a higher MAP has been observed with xenon, when compared with isoflurane anaesthesia. The responses to haemorrhage and subsequent volume replacement were therefore compared between xenon and isoflurane anaesthesia, in pigs. METHODS: Pigs were randomized to anaesthesia with xenon 0.55 MAC (group Xe, n=9) or isoflurane 0.55 MAC (group Iso, n=9), each with remifentanil 0.5 microg kg(-1) min(-1). MAP, heart rate, cardiac output (CO), and left ventricular fractional area change (FAC) were collected at control (1), after haemorrhage (20 ml kg(-1)) (2), after 10 min of recovery (3), after volume replacement (4), and 30 min later (5). Data were analysed by two-way repeated measures anova. RESULTS: Blood loss decreased MAP (Xe: 103 [21] to 53 [24] mm Hg; Iso: 92 [18] to 55 [14] mm Hg) and CO (Xe: 4.1 [0.8] to 2.6 [0.5] litre min(-1); Iso: 5.1 [1.1] to 3.8 [1.2] litre min(-1)), in spite of significant tachycardia. MAP and CO recovered to about 75% of control, and subsequent volume replacement completely reversed symptoms in both groups, but increased FAC only with xenon. CONCLUSION: Haemodynamic response to acute haemorrhage appeared faster with xenon/remifentanil than with isoflurane/remifentanil anaesthesia. In particular MAP decrease and short-term recovery were more marked with xenon (P<0.02). In the xenon group, volume replacement increased FAC compared with control and isoflurane (P<0.02). PMID- 15805141 TI - Recent advances in the non-pharmacological management of postoperative nausea and vomiting. PMID- 15805142 TI - Identifying the sick: can biochemical measurements be used to aid decision making on presentation to the accident and emergency department. AB - BACKGROUND: Early and accurate identification of patients who may benefit from aggressive optimal medical intervention is essential if improved outcomes in terms of survival are to be achieved. We studied the usefulness of routine clinical measurements and/or markers of metabolic abnormality in the early identification of those patients at greatest risk of deterioration on presentation to the accident and emergency department. METHODS: We conducted a prospective observational study in the accident and emergency department of a 602 bed district general hospital. Routine clinical measurements (heart rate, systolic blood pressure, temperature, oxygen saturation in room air, level of consciousness and ventilatory frequency) and venous blood analysis for metabolic markers (pH, bicarbonate, standard base excess, lactate, anion gap, strong ion difference, and strong ion gap) and biochemical markers (Na+, K+, Ca2+, Cl-, PO4- albumin, urea and creatinine) were recorded from unselected consecutive hospital admissions over two 3-month periods (September-November 2002 and February-April 2003). RESULTS: Logistic regression analysis showed that neither conventional clinical measurements upon presentation to the accident and emergency department nor venous biochemical and metabolic indices have good discriminatory ability when used as single predictors of either hospital mortality or length of hospital stay. Selecting variables from all the clinical and venous blood measurements gave a parsimonious model containing only age, heart rate, phosphate and albumin (area under the receiver operating characteristic curve, 0.82 [95% CI 0.76, 0.87]). CONCLUSIONS: A combination of clinical and venous biochemical measurements in the accident and emergency department proved the best predictors of hospital mortality. Consequently, they may be helpful as a triage tool in the accident and emergency department to help identify patients at risk of deterioration. PMID- 15805143 TI - Value of Cushing reflex as warning sign for brain ischaemia during neuroendoscopy. AB - BACKGROUND: During an endoscopic neurosurgical procedure a sudden increase in intracranial pressure may occur at any time. We present a prospective study of haemodynamic changes during such procedures. METHODS: Physiological data were recorded during the whole operative procedure in 17 consecutive patients who underwent an endoscopic neurosurgical procedure under general anaesthesia. Monitoring included invasive blood pressure, intracranial pressure, electrocardiogram, end-expired carbon dioxide, pulse oximetry and heart rate. Pressure and ECG waveforms were recorded at 100 Hz and evaluated in a subsequent offline analysis. RESULTS: In almost every case, the occurrence of hypertension and tachycardia was clearly the result of an increase in intracranial pressure. Also, a Cushing reflex developed in almost every case where the cerebral perfusion pressure dropped below 15 mm Hg. The occurrence of bradycardia was not systematically associated with a low cerebral perfusion pressure. CONCLUSION: In this study, we describe the haemodynamic effects of increased intracranial pressure during endoscopic neurosurgical procedures and their respective sequence of events at high temporal resolution. Although most clinicians rely on the occurrence of bradycardia to diagnose intracranial hypertension during endoscopic neurosurgical procedures, we show that a simultaneous onset of hypertension and tachycardia is a better indicator of impaired brain perfusion. Waiting for a persistent bradycardia to alert the surgeon during endoscopic neurosurgical procedures could allow severe bradycardia or even asystole to develop. PMID- 15805144 TI - Temporary developmental arrest after storage of fertilized mouse oocytes at 4 degrees C: effects on embryonic development, maternal mRNA processing and cell cycle. AB - The aim of this study was to examine whether fertilized mouse oocytes can survive after short-term incubation (for 6-48 h) at 4 degrees C. When fertilized oocytes of ICR and C57BL/6 (B6) strain were incubated at 4 degrees C and returned to normal culture conditions (37 degrees C), development of these 4 degrees C treated embryos for up to 12 h (for ICR) to blastocyst stage did not differ from that of untreated oocytes. Even 4 degrees C-treated embryos for 48 h developed to blastocysts at relatively good rates (33.3% for ICR and 50.8% for B6). The in vivo development of 4 degrees C-treated embryos for 12, 24 and 36 h to fetal stage was similar to that of untreated ones. BrdU labelling assay revealed temporary cessation of DNA replication in 4 degrees C-treated fertilized oocytes. Post-fertilization events including cytoplasmic polyadenylation of maternal mRNAs, mRNA degradation of a cell cycle-related gene and elevated mRNA expression of zygotic gene activation-related genes were temporarily suppressed in 4 degrees C-treated embryos. These findings indicate that 4 degrees C-treatment of fertilized murine oocytes results in temporary cessation of molecular events. We also show that 4 degrees C-treated fertilized oocytes for 12 h can be used for preparation of transgenic mice. PMID- 15805145 TI - Baculovirus-expressed recombinant human zona pellucida glycoprotein-B induces acrosomal exocytosis in capacitated spermatozoa in addition to zona pellucida glycoprotein-C. AB - To facilitate our understanding of the role of zona pellucida glycoproteins during fertilization in humans, recombinant human zona pellucida glycoprotein-A (hZPA), -B (hZPB) and -C (hZPC) were obtained by using Escherichia coli and baculovirus expression systems. Analysis by SDS-PAGE and Western blot of the Ni NTA affinity purified recombinant proteins revealed that the baculovirus expressed hZPA, hZPB and hZPC have an apparent molecular weight of approximately 110, approximately 70-75 and approximately 65 kDa, respectively, as compared to approximately 80, approximately 65 and approximately 50 kDa of the respective E. coli-expressed proteins. Lectin binding studies revealed that the baculovirus expressed recombinant zona proteins were glycosylated. Major oligosaccharides were represented by strong reactivity with Concanavalin A (mannose alpha 1-3 or mannose alpha 1-6 residues) and Jacalin (alpha-O glycosides of Gal or GalNAc moieties). A significant increase in acrosomal exocytosis was observed when capacitated human sperm were incubated in vitro with baculovirus-expressed hZPB (P=0.0005) and hZPC (P=0.0005) The E. coli-expressed hZPB, hZPC and baculovirus expressed hZPA failed to induce any significant increase (P>0.05) in acrosome reaction. In contrast to hZPC, the acrosome reaction induced by recombinant hZPB was not inhibited by pertussis toxin. These studies, for the first time, have demonstrated that in humans, ZPB also induces acrosomal exocytosis through a Gi independent pathway. PMID- 15805146 TI - New enzymatic assay using phospholipase D to measure total calcium in serum. PMID- 15805147 TI - Glutathione S-transferase P1 *C allelic variant increases susceptibility for late onset Alzheimer disease: association study and relationship with apolipoprotein E epsilon4 allele. AB - BACKGROUND: Oxidative stress and neuronal cell death have been implicated in the pathogenesis of Alzheimer disease (AD). Considering that the glutathione transferase (GST) supergene family encodes isoenzymes that appear to be critical in protection against oxidative stress, we aimed at determining the various GSTP1, GSTM1, and GSTT1 polymorphisms and ApoE genotypes to investigate their role as susceptibility genes for late-onset AD (LOAD). METHODS: We included 210 LOAD patients and 228 healthy controls matched for age, sex, and educational level in our case-control genetic association study. GSTM1 and GSTT1 genotypes were studied by conventional PCR, whereas GSTP1 and ApoE genotypes were determined by real-time PCR on the LightCycler. RESULTS: We found a significant association between LOAD and the GSTP1*C allelic variant [odds ratio (OR) = 1.9; P < 0.05], but no association between the GSTM1 and GSTT1 deleted genotypes and LOAD. In addition, a preliminary result suggested that carriers of both the GSTP1*C and ApoE epsilon4 allelic variants were at increased risk of LOAD (OR = 19.98; P < 0.0001). CONCLUSION: The GSTP1*C allelic variant should be considered a candidate for LOAD, particularly in persons having the ApoE epsilon4 allelic variant, because the GSTP1 and ApoE gene products are implicated in oxidative stress and apoptosis processes leading to beta-amyloid-mediated neurodegeneration. PMID- 15805148 TI - A death in the family: reflections on the Terri Schiavo case. PMID- 15805149 TI - Clinical, morphological, and molecular aspects of sialic acid storage disease manifesting in utero. AB - BACKGROUND: Sialic acid storage diseases (SASDs) are caused by the defective transport of free sialic acid outside the lysosome. Apart from the Salla presentation in Finland, SASD is a very rare form of lysosomal storage disease (LSD) with approximately 35 cases, all diagnosed after birth, having been reported worldwide. We report a series of 12 French patients with very early manifestations, including eight fetuses diagnosed in utero. RESULTS: Ultrasound examination, fetal autopsy, or clinical examination showed prominent ascites, rarely progressing to complete hydrops, and highlighted the early severity of bone disease. Dramatic increase of free sialic acid in various biological samples confirmed the diagnosis in all cases. Storage staining affinities and storage distribution in placenta and fetal organs allowed differential diagnosis from other LSDs but cannot differentiate between SASD, sialidosis, and galactosialidosis. Fourteen different mutations were identified, showing the molecular heterogeneity of SASD in the French population. We found that the previously described p.Y306X mutation generated two different transcripts, and we identified seven novel mutations: three deletions (del exon 7, del exons10+11 and c.1296delT), one splice site mutation (c.1350+1G-->T) one nonsense mutation (p.W339X), and two missense mutations (p.R57C and p.G127E). CONCLUSIONS: The severity of our patients' genotypes is in agreement with their phenotypes but not with the importance and early appearance of the very frequent in utero manifestations. Minimal fetal disease in some patients and a reported case of heterogeneity of fetal involvement within a family suggest that factors other than the genotype influence fetal manifestations. PMID- 15805150 TI - Association of the 103I MC4R allele with decreased body mass in 7937 participants of two population based surveys. AB - BACKGROUND: The melanocortin-4-receptor gene (MC4R) is part of the melanocortinergic pathway that controls energy homeostasis. In a recent meta analysis, the MC4R V103I (rs2229616) polymorphism was shown to be associated with body weight regulation. Although no functional differences between the isoleucine comprising receptor and the wild type receptor have been detected as yet, this meta-analysis of 14 case-control studies reported a mild negative association with obesity (odds ratio (OR) 0.69, p = 0.03). However, evidence in a large population based study in a homogeneous population and a significant estimate of the change in quantitative measures of obesity is still lacking. METHODS: We analysed the data of two surveys of a white population with the same high quality study protocol, giving a total of 7937 participants. RESULTS: By linear regression, we found a significant decrease of 0.52 body mass index (BMI) units (95% confidence interval (CI) -0.02 to -1.03, p = 0.043) for carriers of the heterozygote rs2229616G/A genotype, which was observed in 3.7% of the participants. Logistic regression yielded a significantly negative association of the MC4R variant with "above average weight" (BMI > or = median BMI) yielding an OR of 0.75 (95% CI 0.59 to 0.95 p = 0.017). We obtained similar results comparing obese (BMI > or =30 kg/m2, World Health Organization results for 1997) with non obese (BMI < 30 kg/m2) participants. The results were found for both sexes and each survey separately, and did not depend on the modelling of age, sex, or survey effects. CONCLUSIONS: Our study confirms previous findings of a meta analysis that the relatively infrequent G/A genotype of the V103I MC4R polymorphism is negatively associated with above average weight and obesity in population based original data of 7937 participants, and extends previous findings by showing for the first time a significantly lower BMI in individuals carrying the infrequent allele of this MC4R variant. PMID- 15805151 TI - No MSH6 germline mutations in breast cancer families with colorectal and/or endometrial cancer. AB - BACKGROUND: The genetic background in breast cancer families with colorectal and/or endometrial cancer is mostly unknown. The functional connection between MSH6 and the known breast cancer predisposition gene product BRCA1 suggests that the MSH6 gene may also play a role in breast cancer predisposition. METHODS: We analysed 38 breast cancer families with colorectal and/or endometrial cancer for germline mutations in MSH6. RESULTS: No disease associated mutations were detected among the breast cancer families. However, mutation analysis revealed a Glu995STOP mutation in an atypical HNPCC family. The same mutation was found in a patient with both breast and colorectal carcinoma in our previous study, and haplotype analysis confirmed a common ancestral origin. The Glu995STOP mutation was further examined in an extensive series of 245 colorectal and 142 breast carcinoma patients with a family history of breast, colorectal, and/or endometrial carcinoma, and in 268 healthy population controls, but none was found to carry the mutation. CONCLUSIONS: Our results suggest that MSH6 may not be the underlying gene in breast cancer families with a history of colorectal and/or endometrial cancer. The Glu995STOP founder mutation is not a familial breast cancer predisposition allele and makes only a limited contribution to colorectal cancer burden in Finland. PMID- 15805152 TI - High frequency of APOB gene mutations causing familial hypobetalipoproteinaemia in patients of Dutch and Spanish descent. AB - BACKGROUND: Familial hypobetalipoproteinaemia (FHBL) is an autosomal co-dominant hereditary disorder of lipoprotein metabolism characterised by decreased low density lipoprotein (LDL) cholesterol and apolipoprotein B (APOB) plasma levels. High levels of plasma APOB and LDL cholesterol are strong predictors for risk of cardiovascular disease (CVD), while individuals with low APOB and LDL cholesterol levels are thought to have lower than average risk for CVD, and in fact, heterozygous FHBL patients appear to be asymptomatic. METHODS: Rather than identifying truncated APOB proteins in plasma fractions separated by gel electrophoresis, which will miss any mutations in proteins smaller than 30 kb, we analysed the APOB gene directly, using PCR. RESULTS: We identified nine different mutations, six of which are novel. Each mutation showed complete co-segregation with the FHBL phenotype in the families, and statistically significant differences between carriers and non-carriers were found for plasma total, LDL, and HDL cholesterol, triglycerides, and APOB levels, but not for APOA1 levels. All carriers of an APOB mutation were completely free from CVD. CONCLUSIONS: Prolonged low levels of LDL cholesterol and elevated levels of HDL cholesterol may reduce the progression of atherosclerotic disease, but this has not been unequivocally shown that this is indeed the case in individuals with FHBL, and is the subject of a current study. PMID- 15805153 TI - Increased prevalence of imprinting defects in patients with Angelman syndrome born to subfertile couples. AB - Recent case reports have suggested that infertility treatment with intracytoplasmic sperm injection (ICSI) may increase the risk of imprinting defects leading to Angelman syndrome (AS). Although imprinting defects account for only 4% of patients with AS, we have found four cases among 16 AS patients born to subfertile couples, who conceived with or without infertility treatment (25%; relative risk (RR) 6.25; 95% confidence interval (CI) 1.68 to 16.00). The risk in untreated couples with time to pregnancy (TTP) exceeding 2 years was identical to that of those treated by ICSI or by hormonal stimulation alone (RR 6.25; 95% CI 0.70 to 22.57). It was twice as high in couples who had received treatment and also had TTP >2 years (RR 12.5; 95% CI 1.40 to 45.13). Our findings suggest that imprinting defects and subfertility may have a common cause, and that superovulation rather than ICSI may further increase the risk of conceiving a child with an imprinting defect. PMID- 15805154 TI - Distinct CDH3 mutations cause ectodermal dysplasia, ectrodactyly, macular dystrophy (EEM syndrome). AB - BACKGROUND: EEM syndrome is the rare association of ectodermal dysplasia, ectrodactyly, and macular dystrophy. METHODS: We here demonstrate through molecular analysis that EEM is caused by distinct homozygous CDH3 mutations in two previously published families. RESULTS: In family 1, a missense mutation (c.965A-->T) causes a change of amino acid 322 from asparagine to isoleucine; this amino acid is located in a highly conserved motif likely to affect Ca2+ binding affecting specificity of the cell-cell binding function. In family 2, a homozygous frameshift deletion (c.829delG) introduces a truncated fusion protein with a premature stop codon at amino acid residue 295, expected to cause a non functional protein lacking both its intracellular and membrane spanning domains and its extracellular cadherin repeats 3-5. Our mouse in situ expression data demonstrate that Cdh3 is expressed in the apical ectodermal ridge from E10.5 to E12.5, and later in the interdigital mesenchyme, a pattern compatible with the EEM phenotype. Furthermore, we discuss possible explanations for the phenotypic differences between EEM and congenital hypotrichosis with juvenile macular dystrophy (HJMD), which is also caused by CDH3 mutations. CONCLUSIONS: In summary, we have ascertained a third gene associated with ectrodactyly and have demonstrated a hitherto unrecognised role of CDH3 in shaping the human hand. PMID- 15805155 TI - Disruption of the gene Euchromatin Histone Methyl Transferase1 (Eu-HMTase1) is associated with the 9q34 subtelomeric deletion syndrome. AB - BACKGROUND: A new syndrome has been recognised following thorough analysis of patients with a terminal submicroscopic subtelomeric deletion of chromosome 9q. These have in common severe mental retardation, hypotonia, brachycephaly, flat face with hypertelorism, synophrys, anteverted nares, thickened lower lip, carp mouth with macroglossia, and conotruncal heart defects. The minimum critical region responsible for this 9q subtelomeric deletion syndrome (9q-) is approximately 1.2 Mb and encompasses at least 14 genes. OBJECTIVE: To characterise the breakpoints of a de novo balanced translocation t(X;9)(p11.23;q34.3) in a mentally retarded female patient with clinical features similar to the 9q- syndrome. RESULTS: Sequence analysis of the break points showed that the translocation was fully balanced and only one gene on chromosome 9 was disrupted--Euchromatin Histone Methyl Transferase1 (Eu-HMTase1)--encoding a histone H3 lysine 9 methyltransferase (H3-K9 HMTase). This indicates that haploinsufficiency of Eu-HMTase1 is responsible for the 9q submicroscopic subtelomeric deletion syndrome. This observation was further supported by the spatio-temporal expression of the gene. Using tissue in situ hybridisation studies in mouse embryos and adult brain, Eu-HMTase1 was shown to be expressed in the developing nervous system and in specific peripheral tissues. While expression is selectively downregulated in adult brain, substantial expression is retained in the olfactory bulb, anterior/ventral lateral ventricular wall, and hippocampus and weakly in the piriform cortex. CONCLUSIONS: The expression pattern of this gene suggests a role in the CNS development and function, which is in line with the severe mental retardation and behaviour problems in patients who lack one copy of the gene. PMID- 15805156 TI - Multiple mechanisms are implicated in the generation of 5q35 microdeletions in Sotos syndrome. AB - BACKGROUND: Sotos syndrome is characterised by learning difficulties, overgrowth, and a typical facial appearance. Microdeletions at 5q35.3, encompassing NSD1, are responsible for approximately 10% of non-Japanese cases of Sotos. In contrast, a recurrent approximately 2 Mb microdeletion has been reported as responsible for approximately 50% of Japanese cases of Sotos. METHODS: We screened 471 cases for NSD1 mutations and deletions and identified 23 with 5q35 microdeletions. We investigated the deletion size, parent of origin, and mechanism of generation in these and a further 10 cases identified from published reports. We used "in silico" analyses to investigate whether repetitive elements that could generate microdeletions flank NSD1. RESULTS: Three repetitive elements flanking NSD1, designated REPcen, REPmid, and REPtel, were identified. Up to 18 cases may have the same sized deletion, but at least eight unique deletion sizes were identified, ranging from 0.4 to 5 Mb. In most instances, the microdeletion arose through interchromosomal rearrangements of the paternally inherited chromosome. CONCLUSIONS: Frequency, size, and mechanism of generation of 5q35 microdeletions differ between Japanese and non-Japanese cases of Sotos. Our microdeletions were identified from a large case series with a broad range of phenotypes, suggesting that sample selection variability is unlikely as a sole explanation for these differences and that variation in genomic architecture might be a contributory factor. Non-allelic homologous recombination between REPcen and REPtel may have generated up to 18 microdeletion cases in our series. However, at least 15 cannot be mediated by these repeats, including at least seven deletions of different sizes, implicating multiple mechanisms in the generation of 5q35 microdeletions. PMID- 15805157 TI - A homozygous BMPR1B mutation causes a new subtype of acromesomelic chondrodysplasia with genital anomalies. AB - We present a patient with acromesomelic chondrodysplasia and genital anomalies caused by a novel homozygous mutation in BMPR1B, the gene coding for bone morphogenetic protein receptor 1B. The 16 year old girl, the offspring of a multiconsanguinous family, showed a severe form of limb malformation consisting of aplasia of the fibula, severe brachydactyly, ulnar deviation of the hands, and fusion of carpal/tarsal bones. In addition, she presented with hypoplasia of the uterus and ovarian dysfunction resulting in hypergonadotrophic hypogonadism. Mutation analysis of BMPR1B revealed a homozygous 8 bp deletion (del359-366). This mutation is expected to result in a loss of function and is thus different from the heterozygous missense mutations in BMPR1B recently shown to cause brachydactyly type A2 through a dominant negative effect. The patient's skeletal phenotype shows an overlap with the clinical spectrum of the acromesomelic chondrodysplasias of the Grebe, Hunter-Thompson, and DuPan types caused by homozygous mutations in the gene coding for growth differentiation factor 5 (GDF5) which is a high-affinity ligand to BMPR1B. However, the phenotype described here differs from GDF5 associated chondrodysplasias because of the additional presence of genital anomalies and the distinct limb phenotype. PMID- 15805158 TI - Subset of individuals with autism spectrum disorders and extreme macrocephaly associated with germline PTEN tumour suppressor gene mutations. AB - The genetic aetiology of autism remains elusive. Occasionally, individuals with Cowden syndrome (a cancer syndrome) and other related hamartoma disorders such as Bannayan-Riley-Ruvalcaba syndrome, Proteus syndrome, and Proteus-like conditions, are characterised by germline PTEN mutations, and may have neurobehavioural features resembling autism as well as overgrowth and macrocephaly. Therefore, we undertook PTEN gene mutation analysis in 18 subjects mainly prospectively ascertained with autism spectrum disorder and macrocephaly. Of these 18 autistic subjects (13 males and five females; ages 3.1-18.4 years) with a head circumference range from 2.5 to 8.0 standard deviations above the mean, three males (17%) carried germline PTEN mutations. These three probands had previously undescribed PTEN mutations: H93R (exon 4), D252G (exon 7), and F241S (exon 7). They had the larger head circumference measurements amongst all our study subjects. The three residues altered in our patients were highly evolutionarily conserved. We suggest that PTEN gene testing be considered for patients with autistic behaviour and extreme macrocephaly. The gene findings may impact on recurrence risks as well as medical management for the patient. PMID- 15805159 TI - Ancestral RET haplotype associated with Hirschsprung's disease shows linkage disequilibrium breakpoint at -1249. AB - BACKGROUND: Hirschsprung disease (HSCR) is a complex disorder with traditional germline mutations in RET in up to 30% of familial cases and in 3% of sporadic cases in a population-based series. We have previously demonstrated that an ancestral haplotype at the 5' end of RET (haplotype 0) was strongly associated with a large subset of isolated HSCR cases and that a putative low penetrance susceptibility locus was encompassed within this ancestral haplotype, anchored by exon 2 SNP A45A. OBJECTIVE: To determine the 5' extent of the HSCR-associated ancestral haplotype by defining the linkage disequilibrium breakpoint in search for the low penetrance susceptibility locus. METHODS: Systematic screening of the region upstream of the anchoring A45A SNP, comprising RET intron 1, exon 1, and promoter in 117 population-based HSCR cases and 100 controls. Dual luciferase assay to determine differential activities between SNP combinations near a transcription start site. RESULTS: New SNP's were found which formed upstream haplotypes, anchored by A45A, in linkage disequilibrium with HSCR (2 = 76.96, p<0.00000001). Linkage disequilibrium appeared to break at the -1249C/T SNP. Further, the HSCR-associated genotype (00) was found in >60% of HSCR but only 2% of controls. Because only 2 variants, -200A>G and -196C>A, lie within the promoter region and are in proximity to the transcriptional start site (at -195), we modelled these combinations into constructs for luciferase reporter assay. The HSCR-associated SNP combination showed the lowest activity and the control associated combination, the highest. CONCLUSIONS: Our observations seem to discard the existence of a HSCR-causing mutation as it is conceived in the traditional sense, but strengthen the idea of a specific combination of variants conferring susceptibility to the disease in a low penetrance fashion. The data derived from our functional "in vitro" studies would suggest that the HSCR associated haplotype 0 may result in a lower level of expression of the RET gene [corrected] PMID- 15805160 TI - Deletion 9q34.3 syndrome: genotype-phenotype correlations and an extended deletion in a patient with features of Opitz C trigonocephaly. PMID- 15805161 TI - Comprehensive genomic analysis of PKHD1 mutations in ARPKD cohorts. PMID- 15805162 TI - DHCR7 nonsense mutations and characterisation of mRNA nonsense mediated decay in Smith-Lemli-Opitz syndrome. PMID- 15805163 TI - Genetics of Charcot-Marie-Tooth disease type 4A: mutations, inheritance, phenotypic variability, and founder effect. PMID- 15805164 TI - Inadvertent diagnosis of male infertility through genealogical DNA testing. PMID- 15805166 TI - Lipid diffusion, free area, and molecular dynamics simulations. PMID- 15805165 TI - Mean-field model of immobilized enzymes embedded in a grafted polymer layer. AB - Two-dimensional mean-field lattice theory is used to model immobilization and stabilization of an enzyme on a hydrophobic surface using grafted polymers. Although the enzyme affords biofunctionality, the grafted polymers stabilize the enzyme and impart biocompatibility. The protein is modeled as a compact hydrophobic-polar polymer, designed to have a specific bulk conformation reproducing the catalytic cleft of natural enzymes. Three scenarios are modeled that have medical or industrial importance: 1), It is shown that short hydrophilic grafted polymers, such as polyethylene glycol, which are often used to provide biocompatibility, can also serve to protect a surface-immobilized enzyme from adsorption and denaturation on a hydrophobic surface. 2), Screening of the enzyme from the surface and nonspecific interactions with biomaterial in bulk solution requires a grafted layer composed of short hydrophilic polymers and long triblock copolymers. 3), Hydrophilic polymers grafted on a hydrophobic surface in contact with an organic solvent form a dense hydrophilic nanoenvironment near the surface that effectively shields and stabilizes the enzyme against both surface and solvent. PMID- 15805167 TI - Tryptophan substitution of a putative D4S6 gating hinge alters slow inactivation in cardiac sodium channels. AB - Voltage-gated Na(+) channels display rapid activation gating (opening) as well as fast and slow inactivation gating (closing) during depolarization. We substituted residue S1759 (serine), a putative D4S6 gating hinge of human cardiac hNav1.5 Na(+) channels with A (alanine), D (aspartate), K (lysine), L (leucine), P (proline), and W (tryptophan). Significant shifts in gating parameters for activation and steady-state fast inactivation were observed in A-, D-, K-, and W substituted mutant Na(+) channels. No gating shifts occurred in the L-substituted mutant, whereas the P-substituted mutant did not yield sufficient Na(+) currents. Wild-type, A-, D-, and L-substituted mutant Na(+) channels showed little or no slow inactivation with a 10-s conditioning pulse ranging from -180 to 0 mV. Unexpectedly, W- and K-substituted mutant Na(+) channels displayed profound maximal slow inactivation around -100 mV ( approximately 85% and approximately 70%, respectively). However, slow inactivation was progressively reversed in magnitude from -70 to 0 mV. This regression was minimized in inactivation deficient hNav1.5-S1759W/L409C/A410W Na(+) channels, indicating that the intracellular fast-inactivation gate caused such a reversal. Our data suggest that the hNav1.5-S1759 residue plays a critical role in slow inactivation. Possible mechanisms for S1759 involvement in slow inactivation and for antagonism between fast and slow inactivation are discussed. PMID- 15805168 TI - Unraveling protein-protein interactions in living cells with fluorescence fluctuation brightness analysis. AB - Fluorescence correlation spectroscopy is a potentially powerful tool for measuring protein-protein interactions directly in single living cells. We previously reported on the detection of homodimer formation in cells using molecular brightness analysis. Here, we extend the technique to detect binding between different proteins. Proteins are labeled with the fluorescent markers YFP and CFP. We first determine the coexpression ratio of both proteins by measuring the intensity ratio with a dual-color setup. The effect of fluorescence resonance energy transfer on the intensity ratio is explicitly taken into account. The brightness of cells coexpressing both proteins is measured in a single-color setup. Selecting the laser wavelength of the two-photon light source allows us to either coexcite both proteins or to selectively excite YFP-labeled proteins. This approach enables us to distinguish between homodimer and heterodimer formation. We first present the theory and then demonstrate experimental feasibility using the ligand binding domains of retinoic acid receptor (RARLBD) and of retinoid X receptor (RXRLBD). Both proteins form heterodimers, and RXRLBD also forms homodimers in the presence of its agonist. We explore binding between these proteins in the presence and absence of RXR agonist. Our results demonstrate that brightness analysis offers a quantitative method for determining protein interactions in cells. PMID- 15805169 TI - Theoretical IR spectroscopy based on QM/MM calculations provides changes in charge distribution, bond lengths, and bond angles of the GTP ligand induced by the Ras-protein. AB - The GTPase Ras p21 is a crucial switch in cellular signal transduction. Fourier transform infrared (FTIR) spectra of the substrate guanosine triphosphate (GTP) show remarkable changes when it binds to the enzyme. The reduced band widths indicate that the flexible GTP molecule is guided by the protein into a preferred conformation. The delocalized phosphate vibrations of unbound GTP become localized. The frequency shifts show an electron movement toward beta-phosphate, which probably contributes to catalysis by reducing the free activation energy. To quantify these qualitative observations we performed QM/MM molecular dynamics simulations of Ras.GTP and GTP in water. The triphosphate part of GTP was treated quantum mechanically using density functional theory (DFT). Vibrational spectra were calculated in harmonic approximation with an average deviation of 3% from the experimental frequencies. This provides a high confidence in the computational results as vibrational spectra are highly sensitive to conformation and charge distribution. As compared to GTP in water, Ras-bound GTP shows a shift of negative charge of approximately 0.2 e toward the beta-phosphate from gamma phosphate and from alpha-phosphate due to the positive charge of the magnesium ion, to a lesser extent of Lys-16, and surprisingly without any effect of the P loop backbone. Magnesium and Gly-13 twist and bend the gamma-O-beta bonds such that the crucial bond is stretched before cleaving. PMID- 15805170 TI - Nano-sizing of specific gene domains in intact human cell nuclei by spatially modulated illumination light microscopy. AB - Although light microscopy and three-dimensional image analysis have made considerable progress during the last decade, it is still challenging to analyze the genome nano-architecture of specific gene domains in three-dimensional cell nuclei by fluorescence microscopy. Here, we present for the first time chromatin compaction measurements in human lymphocyte cell nuclei for three different, specific gene domains using a novel light microscopic approach called Spatially Modulated Illumination microscopy. Gene domains for p53, p58, and c-myc were labeled by fluorescence in situ hybridization and the sizes of the fluorescence in situ hybridization "spots" were measured. The mean diameters of the gene domains were determined to 103 nm (c-myc), 119 nm (p53), and 123 nm (p58) and did not correlate to the genomic, labeled sequence length. Assuming a spherical domain shape, these values would correspond to volumes of 5.7 x 10(-4) microm(3) (c-myc), 8.9 x 10(-4) microm(3) (p53), and 9.7 x 10(-4) microm(3) (p58). These volumes are approximately 2 orders of magnitude smaller than the diffraction limited illumination or observation volume, respectively, in a confocal laser scanning microscope using a high numerical aperture objective lens. By comparison of the labeled sequence length to the domain size, compaction ratios were estimated to 1:129 (p53), 1:235 (p58), and 1:396 (c-myc). The measurements demonstrate the advantage of the SMI technique for the analysis of gene domain nano-architecture in cell nuclei. The data indicate that chromatin compaction is subjected to a large variability which may be due to different states of genetic activity or reflect the cell cycle state. PMID- 15805171 TI - A stopped-flow kinetic study of the assembly of nonviral gene delivery complexes. AB - Stopped-flow circular dichroism and fluorescence spectroscopy are used to characterize the assembly of complexes consisting of plasmid DNA bound to the cationic lipids dimethyldioctadecylammonium bromide and 1, 2-dioleoyl- 3 trimethylammonium-propane and a series of polyamidoamine dendrimers. The kinetics of complexation determined from the stopped-flow circular dichroism measurements suggests complexation occurs within 50 ms. Further analysis, however, was precluded by the presence of mixing (shear) artifacts. Stopped-flow fluorescence employing the high-affinity DNA dyes Hoechst 33258 and YOYO-1 was able to resolve two sequential steps in the assembly of complexes that are assigned to binding/dehydration and condensation events. The rates of each process were determined over the temperature range of 10-50 degrees C and activation energies were determined from the slope of Arrhenius plots. The behavior of polyamidoamine dendrimers can be separated into two classes based on their differing binding modes: generation 2 and the larger generations (G4, G7, and G9). The larger generations have activation energies for binding that follow the trend G4 > G7 > G9. The activation energies for condensation (compaction) of complexes composed of these same dendrimers have the opposite trend G9 > G7 > G4. It is postulated that a balance between a more energetically favorable condensation and less favorable binding may prove beneficial in enhancing gene delivery. PMID- 15805172 TI - Chiral differentiation of DNA adducts formed by enantiomeric analogues of antitumor cisplatin is sequence-dependent. AB - 1,2-GG intrastrand cross-links formed in DNA by the enantiomeric complexes [PtCl(2)(R,R-2,3-diaminobutane (DAB))] and [PtCl(2)(S,S-DAB)] were studied by biophysical methods. Molecular modeling revealed that structure of the cross links formed at the TGGT sequence was affected by repulsion between the 5' directed methyl group of the DAB ligand and the methyl group of the 5'-thymine of the TGGT fragment. Molecular dynamics simulations of the solvated platinated duplexes and our recent structural data indicated that the adduct of [PtCl(2)(R,R DAB)] alleviated this repulsion by unwinding the TpG step, whereas the adduct of [PtCl(2)(S,S-DAB)] avoided the unfavorable methyl-methyl interaction by decreasing the kink angle. Electrophoretic retardation measurements on DNA duplexes containing 1,2-GG intrastrand cross-links of Pt(R,R-DAB)(2+) or Pt(S,S DAB)(2+) at a CGGA site showed that in this sequence both enantiomers distorted the double helix to the identical extent similar to that found previously for the same sequence containing the cross-links of the parent antitumor cis Pt(NH(3))(2)(2+) (cisplatin). In addition, the adducts showed similar affinities toward the high-mobility-group box 1 proteins. Hence, whereas the structural perturbation induced in DNA by 1,2-GG intrastrand cross-links of cisplatin does not depend largely on the bases flanking the cross-links, the perturbation related to GG cross-linking by bulkier platinum diamine derivatives does. PMID- 15805173 TI - A coarse-grained molecular model for glycosaminoglycans: application to chondroitin, chondroitin sulfate, and hyaluronic acid. AB - A coarse-grained molecular model is presented for the study of the equilibrium conformation and titration behavior of chondroitin (CH), chondroitin sulfate (CS), and hyaluronic acid (HA)-glycosaminoglycans (GAGs) that play a central role in determining the structure and biomechanical properties of the extracellular matrix of articular cartilage. Systematic coarse-graining from an all-atom description of the disaccharide building blocks retains the polyelectrolytes' specific chemical properties while enabling the simulation of high molecular weight chains that are inaccessible to all-atom representations. Results are presented for the characteristic ratio, the ionic strength-dependent persistence length, the pH-dependent expansion factor for the end-to-end distance, and the titration behavior of the GAGs. Although 4-sulfation of the N-acetyl-D galactosamine residue is found to increase significantly the intrinsic stiffness of CH with respect to 6-sulfation, only small differences in the titration behavior of the two sulfated forms of CH are found. Persistence length expressions are presented for each type of GAG using a macroscopic (wormlike chain-based) and a microscopic (bond vector correlation-based) definition. Model predictions agree quantitatively with experimental conformation and titration measurements, which support use of the model in the investigation of equilibrium solution properties of GAGs. PMID- 15805174 TI - DNA packaging in bacteriophage: is twist important? AB - We study the packaging of DNA into a bacteriophage capsid using computer simulation, specifically focusing on the potential impact of twist on the final packaged conformation. We perform two dynamic simulations of packaging a polymer chain into a spherical confinement: one where the chain end is rotated as it is fed, and one where the chain is fed without end rotation. The final packaged conformation exhibits distinct differences in these two cases: the packaged conformation from feeding with rotation exhibits a spool-like character that is consistent with experimental and previous theoretical work, whereas feeding without rotation results in a folded conformation inconsistent with a spool conformation. The chain segment density shows a layered structure, which is more pronounced for packaging with rotation. However, in both cases, the conformation is marked by frequent jumps of the polymer chain from layer to layer, potentially influencing the ability to disentangle during subsequent ejection. Ejection simulations with and without Brownian forces show that Brownian forces are necessary to achieve complete ejection of the polymer chain in the absence of external forces. PMID- 15805175 TI - Cross-polarized reflected light measurement of fast optical responses associated with neural activation. AB - We developed an optical probe for cross-polarized reflected light measurements and investigated optical signals associated with electrophysiological activation in isolated lobster nerves. The cross-polarized baseline light intensity (structural signal) and the amplitude of the transient response to stimulation (functional signal) measured in reflected mode were dependent on the orientation of the nerve axis relative to the polarization plane of incident light. The maximum structural signal and functional response amplitude were observed at 45 degrees , and the ratio of functional to structural signals was approximately constant across orientations. Functional responses were measured in single trials in both transmitted and reflected geometries and responses had similar waveforms. Both structural and functional signals were an order of magnitude smaller in reflected than in transmitted light measurements, but functional responses had similar signal/noise ratios. We propose a theoretical model based on geometrical optics that is consistent with experimental results. In the model, the cross polarized structural signal results from light reflection from axonal fibers and the transient functional response arises from axonal swelling associated with neural activation. Polarization-sensitive reflected light measurements could greatly enhance in vivo imaging of neural activation since cross-polarized responses are much larger than scattering signals now employed for dynamic functional neuroimaging. PMID- 15805176 TI - The emergence of scaling in sequence-based physical models of protein evolution. AB - It has recently been discovered that many biological systems, when represented as graphs, exhibit a scale-free topology. One such system is the set of structural relationships among protein domains. The scale-free nature of this and other systems has previously been explained using network growth models that, although motivated by biological processes, do not explicitly consider the underlying physics or biology. In this work we explore a sequence-based model for the evolution protein structures and demonstrate that this model is able to recapitulate the scale-free nature observed in graphs of real protein structures. We find that this model also reproduces other statistical feature of the protein domain graph. This represents, to our knowledge, the first such microscopic, physics-based evolutionary model for a scale-free network of biological importance and as such has strong implications for our understanding of the evolution of protein structures and of other biological networks. PMID- 15805177 TI - Normal mode analysis suggests a quaternary twist model for the nicotinic receptor gating mechanism. AB - We present a three-dimensional model of the homopentameric alpha7 nicotinic acetylcholine receptor (nAChR), that includes the extracellular and membrane domains, developed by comparative modeling on the basis of: 1), the x-ray crystal structure of the snail acetylcholine binding protein, an homolog of the extracellular domain of nAChRs; and 2), cryo-electron microscopy data of the membrane domain collected on Torpedo marmorata nAChRs. We performed normal mode analysis on the complete three-dimensional model to explore protein flexibility. Among the first 10 lowest frequency modes, only the first mode produces a structural reorganization compatible with channel gating: a wide opening of the channel pore caused by a concerted symmetrical quaternary twist motion of the protein with opposing rotations of the upper (extracellular) and lower (transmembrane) domains. Still, significant reorganizations are observed within each subunit, that involve their bending at the domain interface, an increase of angle between the two beta-sheets composing the extracellular domain, the internal beta-sheet being significantly correlated to the movement of the M2 alpha-helical segment. This global symmetrical twist motion of the pentameric protein complex, which resembles the opening transition of other multimeric ion channels, reasonably accounts for the available experimental data and thus likely describes the nAChR gating process. PMID- 15805178 TI - Moraxella catarrhalis in chronic obstructive pulmonary disease: burden of disease and immune response. AB - RATIONALE: Moraxella catarrhalis is frequently present in the sputum of adults with chronic obstructive pulmonary disease (COPD). Little is known about the role of M. catarrhalis in this common disease. OBJECTIVE: To elucidate the burden of disease, the dynamics of carriage, and immune responses to M. catarrhalis in COPD. METHODS: Prospective cohort study of 104 adults with COPD in an outpatient clinic at the Buffalo Veterans Affairs Medical Center. MEASUREMENTS: Clinical information, sputum cultures, molecular typing of isolates, and immunoassays to measure antibodies to M. catarrhalis. MAIN RESULTS: Over 81 months, 104 patients made 3,009 clinic visits, 560 during exacerbations. Molecular typing identified 120 episodes of acquisition and clearance of M. catarrhalis in 50 patients; 57 (47.5%) of the acquisitions were associated with clinical exacerbations. No instances of simultaneous acquisition of a new strain of another pathogen were observed. The duration of carriage of M. catarrhalis was shorter with exacerbations compared with asymptomatic colonization (median, 31.0 vs. 40.4 days; p = 0.01). Reacquisition of the same strain was rare. The intensity of the serum IgG response was greater after exacerbations than asymptomatic colonization (p = 0.009). Asymptomatic colonization was associated with a greater frequency of a sputum IgA response than exacerbation (p = 0.009). CONCLUSIONS: M. catarrhalis likely causes approximately 10% of exacerbations of COPD, accounting for approximately 2 to 4 million episodes annually. The organism is cleared efficiently after a short duration of carriage. Patients develop strain-specific protection after clearance of M. catarrhalis from the respiratory tract. PMID- 15805179 TI - Sex differences in factors associated with childhood- and adolescent-onset wheeze. AB - RATIONALE: Factors predicting the development of wheeze may differ between sexes and between childhood and adolescence. METHODS: A New Zealand birth cohort of 1,037 children was followed to age 26. For this analysis, those reporting recurrent wheezing at two or more assessments were classified as "wheezers." We examined risk factors for development of wheeze before age 10 (childhood) and subsequently (adolescent-onset) for males and for females separately using Cox regression modeling. RESULTS: Males more often developed childhood wheeze (p = 0.002) and females adolescent-onset wheeze (p < 0.001). Maternal atopy (asthma or hay fever) was a risk factor for childhood wheeze in both sexes (hazard ratio [HR], 1.48, p < 0.05 for males; HR, 2.37, p < 0.001 for females). Paternal atopy also influenced childhood wheeze, significantly for males (HR, 1.72; p = 0.01), and similarly but not significantly for females (HR, 1.70; p = 0.08). For adolescent-onset wheeze, neither maternal (HR, 1.41; p = 0.19) nor paternal history (HR, 0.73; p = 0.42) was a risk factor in males, but maternal history (HR, 2.08; p < 0.01) was a significant risk factor for females. When both age ranges were combined, providing greater power for analysis, paternal history was a stronger risk factor for wheeze in females (HR, 1.62; p = 0.02) than in males (HR, 1.35; p = 0.12). CONCLUSION: The influence of parental atopy on the development for wheeze differs between males and females and between childhood- and adolescent-onset wheeze. PMID- 15805180 TI - Polymorphisms in a disintegrin and metalloprotease 33 (ADAM33) predict impaired early-life lung function. AB - RATIONALE: Asthma commonly originates in early life in association with impaired lung function, which tracks to adulthood. OBJECTIVES: Within the context of a prospective birth cohort study, we investigated the association between single nucleotide polymorphisms (SNPs) in a disintegrin and metalloprotease 33 (ADAM33) gene and early-life lung function. METHODS: Children were genotyped for 17 SNPs in ADAM33. Lung function at age 3 (n = 285) and 5 years (n = 470) was assessed using plethysmographic measurement of specific airway resistance (sRaw). At age 5, we also measured FEV(1). SNPs were analyzed individually using logistic regression, followed by linkage disequilibrium mapping to identify the causal locus. MAIN RESULTS: Carriers of the rare allele of F+1 SNP had reduced lung function at age 3 years (p = 0.003). When the recessive model was considered, four SNPs (F+1, S1, ST+5, V4) showed association with sRaw at age 5 years (p < 0.04). Using linkage disequilibrium mapping, we found evidence of a significant causal location between BC+1 and F1 SNPs, at the 5' end of the gene. Four SNPs were associated with lower FEV(1) (F+1, M+1, T1, and T2; p < or = 0.04). The risk of transient early wheezing more than doubled among children homozygous for the A allele of F+1 (odds ratio, 2.39; 95% confidence intervals, 1.18-4.86; p = 0.02), but there was no association between any SNP and allergic sensitization or physician-diagnosed asthma. CONCLUSIONS: Polymorphisms in ADAM33 predict impaired early-life lung function. The functionally relevant polymorphism is likely to be at the 5' end of the gene. PMID- 15805181 TI - Haemophilus influenzae from patients with chronic obstructive pulmonary disease exacerbation induce more inflammation than colonizers. AB - RATIONALE: Airway infection with Haemophilus influenzae causes airway inflammation, and isolation of new strains of this bacteria is associated with increased risk of exacerbations in patients with chronic obstructive pulmonary disease (COPD). OBJECTIVE: To determine whether strains of H. influenzae associated with exacerbations cause more inflammation than strains that colonize the airways of patients with COPD. METHODS: Exacerbation strains of H. influenzae were isolated from patients during exacerbation of clinical symptoms with subsequent development of a homologous serum antibody response and were compared with colonization strains that were not associated with symptom worsening or an antibody response. Bacterial strains were compared using an in vivo mouse model of airway infection and in vitro cell culture model of bacterial adherence and defense gene and signaling pathway activation in primary human airway epithelial cells. RESULTS: H. influenzae associated with exacerbations caused more airway neutrophil recruitment compared with colonization strains in the mouse model of airway bacterial infection. Furthermore, exacerbation strains adhered to epithelial cells in significantly higher numbers and induced more interleukin-8 release after interaction with airway epithelial cells. This effect was likely mediated by increased activation of the nuclear factor-kappaB and p38 mitogen activated protein kinase signaling pathways. CONCLUSIONS: The results indicate that H. influenzae strains isolated from patients during COPD exacerbations often induce more airway inflammation and likely have differences in virulence compared with colonizing strains. These findings support the concept that bacteria infecting the airway during COPD exacerbations mediate increased airway inflammation and contribute to decreased airway function. PMID- 15805182 TI - The early bactericidal activities of rifampin and rifapentine in pulmonary tuberculosis. AB - RATIONALE: Comparison of the early bactericidal activity (EBA) of rifapentine and its pharmacokinetics with those of rifampin to determine the cause of poor clinical response and regrowth between doses, leading to rifamycin monoresistance at relapse. OBJECTIVES: Determination of the dose size of rifapentine that gives sufficient drug exposure to prevent regrowth. METHODS: EBA study over initial 5 days of treatment of 123 patients, half at Durban and half at Cape Town, who received single rifapentine doses of 300, 600, 900, or 1,200 mg rifapentine or five daily doses of 150, 300, or 600 mg rifampin, with a pharmacokinetic study on 58 patients measuring standard parameters for each dose size of rifamycin and their desacetyl metabolites. RESULTS: The EBAs for both rifamycins were similar, with a linear relationship to log dose at lower doses and a curvilinear response at higher doses giving a plateau at 1,136 mg rifapentine. The area under the concentration-time curve (AUC) divided by the minimal inhibitory concentration (MIC) agreed well for both rifamycins on the assumption that the only free 2% of free rifapentine and the 14% of free rifampin after plasma binding were active in the lesions. CONCLUSIONS: Only the free proportions of the rifamycins were active in lesions. From consideration of the pulse size and the duration of the postantibiotic lag, a 1,200-mg dose of rifapentine seemed necessary to improve response and to prevent regrowth between doses, and hence rifamycin monoresistance. PMID- 15805184 TI - Risk factors for active tuberculosis after antiretroviral treatment initiation in Abidjan. AB - RATIONALE: In sub-Saharan Africa: (1) tuberculosis is the first cause of HIV related mortality; (2) the incidence of tuberculosis in adults receiving highly active antiretroviral therapy (HAART) is lower than in untreated HIV-infected adults but higher than in HIV-negative adults; and (3) factors associated with the occurrence of tuberculosis in patients receiving HAART have never been described. OBJECTIVE: To look for the risk factors for active tuberculosis in HIV infected adults receiving HAART in Abidjan. METHODS: Seven-year prospective cohort of HIV-infected adults, with standardized procedures for documenting morbidity. We analyzed the incidence of active tuberculosis in patients who started HAART and the association between the occurrence of tuberculosis and the characteristics of these patients at HAART initiation. MAIN RESULTS: A total of 129 adults (median baseline CD4 count 125/mm(3)) started HAART and were then followed for 270 person-years (P-Y). At HAART initiation, 31 had a history of tuberculosis and none had current active tuberculosis. During follow-up, the incidence of active tuberculosis was 4.8/100 P-Y (95% confidence interval [CI], 2.5-8.3) overall, 3.0/100 P-Y (95% CI, 1.1-6.6) in patients with no tuberculosis history, and 11.3/100 P-Y (95% CI, 4.1-24.5) in patients with a history of tuberculosis (adjusted hazard ratio, 4.64; 95% CI, 1.29-16.62, p = 0.02). CONCLUSION: The risk of tuberculosis after HAART initiation was significantly higher in patients with a history of tuberculosis than in those with no tuberculosis history. If confirmed by others, this finding could lead to assessment of new patterns of time-limited tuberculosis secondary chemoprophylaxis during the period of initiation of HAART in sub-Saharan African adults. PMID- 15805185 TI - Steroid-resistant inflammation in a rat model of chronic obstructive pulmonary disease is associated with a lack of nuclear factor-kappaB pathway activation. AB - RATIONALE: Emphysema is one component of chronic obstructive pulmonary disease (COPD), a respiratory disease currently increasing in prevalence worldwide. The mainstay therapy adopted to treat patients with COPD is glucocorticoids; unfortunately, this treatment has limited impact on disease symptoms or underlying airway inflammation. OBJECTIVE: There is an urgent need to develop therapies that modify both the underlying inflammation, thought to be involved in disease progression, and the structural changes in the emphysematous lung. METHODS: We have characterized an elastase-driven model of experimental emphysema in the rat that demonstrates COPD-like airway inflammation and determined the impact of a clinically relevant glucocorticoid. MEASUREMENTS AND MAIN RESULTS: We observed an increase in lung neutrophils, lymphomononuclear cells, mucus production, and inflammatory cytokines. Also present were increases in average air space area, which are associated with emphysema-like changes in lung function, such as increased residual volume and decreased flow; these increases in area were maintained for up to 10 weeks. In addition, we observed that elastase-induced airway neutrophilia is steroid resistant. Interestingly, the inflammation observed after elastase administration was found to be temporally associated with a lack of nuclear factor-kappaB pathway activation. This apparent nuclear factor-kappaB-independent inflammation may explain why treatment with a glucocorticoid was ineffective in this preclinical model and could suggest parallels in the steroid-resistant human disease. CONCLUSION: We believe that this model, in addition to its suitability for testing therapies that may modify existing emphysema, could be useful in the search for new therapies to reduce the steroid-resistant airway inflammation evident in COPD. PMID- 15805183 TI - Tumor necrosis factor-alpha overexpression in lung disease: a single cause behind a complex phenotype. AB - RATIONALE: Tumor necrosis factor alpha (TNF-alpha) has been implicated as a key cytokine in many inflammatory lung diseases. These effects are currently unclear, because a transgenic mouse overexpressing TNF-alpha in the lung has been shown in separate studies to produce elements of both emphysema and pulmonary fibrosis. OBJECTIVES: We sought to elucidate the phenotypic effects of TNF-alpha overexpression in a mouse model. MEASUREMENTS: We established the phenotype by measuring lung impedance and thoracic gas volume, and using micro-computed tomography and histology. MAIN RESULTS: We found that airways resistance in this mouse was not different to control mice, but that lung tissue dampening, elastance, and hysteresivity were significantly elevated. Major heterogeneous abnormalities of the parenchyma were also apparent in histologic sections and in micro-computed tomography images of the lung. These changes included airspace enlargement, loss of small airspaces, increased collagen, and thickened pleural septa. We also found significant increases in lung and chest cavity volumes in the TNF-alpha-overexpressing mice. CONCLUSIONS: We conclude that TNF-alpha overexpression causes pathologic changes consistent with both emphysema and pulmonary fibrosis combined with a general lung inflammation, and consequently does not model any single human disease. Our study thus confirms the pleiotropic effects of TNF-alpha, which has been implicated in multiple inflammatory disorders, and underscores the necessity of using a wide range of investigative techniques to link gene expression and phenotype in animal models of disease. PMID- 15805186 TI - The adult incidence of asthma and respiratory symptoms by passive smoking in uterus or in childhood. AB - The effects of pre- or postnatal passive smoking on the adult incidence of asthma have not been reported previously. Between 1985 and 1996/1997, we conducted an 11 year community cohort study on the incidence of asthma and respiratory symptoms in Western Norway. The cohort included 3,786 subjects aged 15 to 70 years, of which 2,819 were responders at both baseline and follow-up. The incidence of asthma and five respiratory symptoms by self-reported exposure to maternal smoking in utero and in childhood, as well as smoking by other household members in childhood, was examined. After adjustment for sex, age, education, hay fever, personal smoking, and occupational exposure, maternal smoking was associated with asthma, phlegm cough, chronic cough, dyspnea grade 2, attacks of dyspnea, and wheezing, with odds ratios (95% confidence intervals [CI]) of 3.0 (1.6, 5.6), 1.7 (1.1, 2.6), 1.9 (1.2, 3.0), 1.9 (1.2, 3.0), 2.0 (1.3, 3.0), and 1.4 (0.9, 2.2), respectively. The adjusted attributable fractions (95% CI) of the adult incidence of asthma were 17.3% (5.2, 27.9) caused by maternal smoking and 9.3% (95% CI, 23.2, 33.2) caused by smoking by other household members. Exposure to pre- and postnatal smoking carries a substantial risk for developing adult asthma and respiratory symptoms. PMID- 15805187 TI - Clinical relevance of Mycobacterium tuberculosis plcD gene mutations. AB - To identify Mycobacterium tuberculosis virulence factors, we integrated comparative genomics and epidemiologic data analysis to investigate the relationship between certain genomic insertions and deletions in the phospholipase-C gene D (plcD) with the clinical presentation of tuberculosis (TB). Four hundred ninety-six well-characterized M. tuberculosis clinical isolates were studied. Approximately 30% (147) of the isolates had an interruption of the plcD gene. Patients infected with the plcD mutant were twice as likely to have extrathoracic disease as those infected by a strain without an interruption (adjusted odds ratio, 2.19; 95% confidence interval, 1.27, 3.76). When we limited the analysis to the 275 isolates with distinct DNA fingerprint patterns, we observed the same association (adjusted odds ratio, 2.74; 95% confidence interval, 1.35, 5.56). Furthermore, the magnitude of the association appeared to differ with the type of extrathoracic TB. Our findings suggest that the plcD gene of M. tuberculosis is potentially involved in the pathogenesis of TB, and the clinical presentation of the disease may be influenced by the genetic variability of the plcD region. PMID- 15805188 TI - Nonsense mutations in folliculin presenting as isolated familial spontaneous pneumothorax in adults. AB - RATIONALE: Approximately 10% of patients who have a spontaneous pneumothorax have a positive family history. OBJECTIVES: We sought to identify DNA sequence variations that confer susceptibility to pneumothoraces. METHODS: We collected 12 families that had at least 2 first-degree relatives with a spontaneous pneumothorax. All affected family members had no obvious stigmata of known genetic disorders associated with pneumothoraces. We used haplotype analysis, DNA sequencing, and restriction fragment analysis of mutations to evaluate the individuals in these families. MAIN RESULTS: In 2 of the 12 families the disorder cosegregated with markers flanking a candidate locus, FLCN. Sequencing the linked alleles revealed 2 mutations predicted to introduce premature stop codons in 2 of the 12 families. Most mutations in FLCN cause a rare disease, Birt-Hogg-Dube syndrome, characterized by autosomal dominant inheritance of multiple benign skin lesions, renal tumors, pulmonary blebs, and pneumothoraces. None of the family members with the nonsense mutations had the skin manifestations of Birt-Hogg-Dube syndrome or renal cancer. Pathologic examination of lung tissue from three affected nonsmokers revealed blebs and underlying emphysema. CONCLUSIONS: Isolated familial spontaneous pneumothorax can be caused by mutations of the FLCN gene. Because development of a pneumothorax and/or pulmonary blebs may be the earliest or the only clinical manifestation of FLCN mutations, pulmonologists should be alert to the contribution of this gene toward this familial form of emphysema. PMID- 15805189 TI - Plakoglobin suppresses keratinocyte motility through both cell-cell adhesion dependent and -independent mechanisms. AB - Plakoglobin (PG) is a member of the Armadillo family of adhesion/signaling proteins and has been shown to play a critical role in the organization of desmosomes and tissue integrity. Because dissolution of intercellular junctions is frequently an initial step in the onset of epithelial cell migration, we examined whether loss of PG promotes cell motility by compromising adhesive strength. Keratinocyte cultures established from PG-/-mice exhibited weakened adhesion and increased motility in transwell migration assays; both were restored by reintroducing PG through adenoviral infection. Interestingly, single PG-/- cells also exhibited increased motility, which was suppressed by reintroducing PG, but not the closely related beta-catenin. Whereas both N- and C-terminally truncated PG deletion mutants restored adhesion, only N-terminally deleted PG, but not C-terminally deleted PG, suppressed single-cell migration. Furthermore, both the chemical inhibitor PP2 and dominant-negative Src tyrosine kinase inhibited single-cell motility in PG-/- cells, whereas constitutively active Src overcame the inhibitory effect of PG. These data demonstrate that PG strengthens adhesion and suppresses motility in mouse keratinocytes, through both intercellular adhesion-dependent and -independent mechanisms, the latter of which may involve suppression of Src signaling through a mechanism requiring the PG C terminus. PMID- 15805190 TI - Decreased plasma cholesterol and hypersensitivity to statins in mice lacking Pcsk9. AB - PCSK9 encodes proprotein convertase subtilisin/kexin type 9a (PCSK9), a member of the proteinase K subfamily of subtilases. Missense mutations in PCSK9 cause an autosomal dominant form of hypercholesterolemia in humans, likely due to a gain of-function mechanism because overexpression of either WT or mutant PCSK9 reduces hepatic LDL receptor protein (LDLR) in mice. Here, we show that livers of knockout mice lacking PCSK9 manifest increased LDLR protein but not mRNA. Increased LDLR protein led to increased clearance of circulating lipoproteins and decreased plasma cholesterol levels (46 mg/dl in Pcsk9(-/-) mice versus 96 mg/dl in WT mice). Statins, a class of drugs that inhibit cholesterol synthesis, increase expression of sterol regulatory element-binding protein-2 (SREBP-2), a transcription factor that activates both the Ldlr and Pcsk9 genes. Statin administration to Pcsk9(-/-) mice produced an exaggerated increase in LDLRs in liver and enhanced LDL clearance from plasma. These data demonstrate that PCSK9 regulates the amount of LDLR protein in liver and suggest that inhibitors of PCSK9 may act synergistically with statins to enhance LDLRs and reduce plasma cholesterol. PMID- 15805191 TI - Domain III of Plasmodium falciparum apical membrane antigen 1 binds to the erythrocyte membrane protein Kx. AB - Plasmodium falciparum apical membrane antigen 1 (AMA1) is located in the merozoite micronemes, an organelle that contains receptors for invasion, suggesting that AMA1 may play a role in this process. However, direct evidence that P. falciparum AMA1 binds to human erythrocytes is lacking. In this study, we determined that domain III of AMA1 binds to the erythrocyte membrane protein, Kx, and that the rate of invasion of Kx(null) erythrocytes is reduced, indicating a significant but not unique role of AMA1 and Kx in parasite invasion of erythrocytes. Domains I/II/III, domains I/II and domain III of AMA1 were expressed on the surface of CHO-K1 cells, and their ability to bind erythrocytes was determined. We observed that each of these domains failed to bind untreated human erythrocytes. In contrast, domain III, but not the other domains of AMA1, bound to trypsin-treated human erythrocytes. We tested the binding of AMA1 to trypsin-treated genetically mutant human erythrocytes, missing various erythrocyte membrane proteins. AMA1 failed to bind trypsin-treated Kx(null) (McLeod) erythrocytes, which lack the Kx protein. Furthermore, treatment of human erythrocytes with trypsin, followed by alpha-chymotrypsin, cleaved Kx and destroyed the binding of AMA1 to human erythrocytes. Lastly, the rate of invasion of Kx null erythrocytes by P. falciparum was significantly lower than Kx expressing erythrocytes. Taken together, our data suggest that AMA1 plays an important, but not exclusive, role in invasion of human erythrocytes through a process that involves exposure or modification of the erythrocyte surface protein, Kx, by a trypsin-like enzyme. PMID- 15805192 TI - Functional diffusion map: a noninvasive MRI biomarker for early stratification of clinical brain tumor response. AB - Assessment of radiation and chemotherapy efficacy for brain cancer patients is traditionally accomplished by measuring changes in tumor size several months after therapy has been administered. The ability to use noninvasive imaging during the early stages of fractionated therapy to determine whether a particular treatment will be effective would provide an opportunity to optimize individual patient management and avoid unnecessary systemic toxicity, expense, and treatment delays. We investigated whether changes in the Brownian motion of water within tumor tissue as quantified by using diffusion MRI could be used as a biomarker for early prediction of treatment response in brain cancer patients. Twenty brain tumor patients were examined by standard and diffusion MRI before initiation of treatment. Additional images were acquired 3 weeks after initiation of chemo- and/or radiotherapy. Images were coregistered to pretreatment scans, and changes in tumor water diffusion values were calculated and displayed as a functional diffusion map (fDM) for correlation with clinical response. Of the 20 patients imaged during the course of therapy, 6 were classified as having a partial response, 6 as stable disease, and 8 as progressive disease. The fDMs were found to predict patient response at 3 weeks from the start of treatment, revealing that early changes in tumor diffusion values could be used as a prognostic indicator of subsequent volumetric tumor response. Overall, fDM analysis provided an early biomarker for predicting treatment response in brain tumor patients. PMID- 15805193 TI - Genetic predictors of the maximum doses patients receive during clinical use of the anti-epileptic drugs carbamazepine and phenytoin. AB - Phenytoin and carbamazepine are effective and inexpensive anti-epileptic drugs (AEDs). As with many AEDs, a broad range of doses is used, with the final "maintenance" dose normally determined by trial and error. Although many genes could influence response to these medicines, there are obvious candidates. Both drugs target the alpha-subunit of the sodium channel, encoded by the SCN family of genes. Phenytoin is principally metabolized by CYP2C9, and both are probable substrates of the drug transporter P-glycoprotein. We therefore assessed whether variation in these genes associates with the clinical use of carbamazepine and phenytoin in cohorts of 425 and 281 patients, respectively. We report that a known functional polymorphism in CYP2C9 is highly associated with the maximum dose of phenytoin (P = 0.0066). We also show that an intronic polymorphism in the SCN1A gene shows significant association with maximum doses in regular usage of both carbamazepine and phenytoin (P = 0.0051 and P = 0.014, respectively). This polymorphism disrupts the consensus sequence of the 5' splice donor site of a highly conserved alternative exon (5N), and it significantly affects the proportions of the alternative transcripts in individuals with a history of epilepsy. These results provide evidence of a drug target polymorphism associated with the clinical use of AEDs and set the stage for a prospective evaluation of how pharmacogenetic diagnostics can be used to improve dosing decisions in the use of phenytoin and carbamazepine. Although the case made here is compelling, our results cannot be considered definitive or ready for clinical application until they are confirmed by independent replication. PMID- 15805194 TI - A checkpoint control linking meiotic S phase and recombination initiation in fission yeast. AB - During meiosis, high levels of recombination initiated by DNA double-strand breaks (DSBs) occur only after DNA replication. However, how DSB formation is coupled to DNA replication is unknown. We examined several DNA replication proteins for a role in this coupling in Schizosaccharomyces pombe, and we show that ribonucleotide reductase, the rate-limiting enzyme of deoxyribonucleotide synthesis and the target of the DNA synthesis inhibitor hydroxyurea (HU) is indirectly required for DSB formation linked to DNA replication. However, in cells in which the function of the DNA-replication-checkpoint proteins Rad1p, Rad3p, Rad9p, Rad17p, Rad26p, Hus1p, or Cds1p was compromised, DSB formation occurred at similar frequencies in the absence or presence of HU. The DSBs in the HU-treated mutant cells occurred at normal sites and were associated with recombination. In addition, Cdc2p is apparently not involved in this process. We propose that the sequence of meiotic S phase and initiation of recombination is coordinated by DNA-replication-checkpoint proteins. PMID- 15805195 TI - "Minimally invasive" total hip arthroplasty. PMID- 15805196 TI - A minimal-incision technique in total hip arthroplasty does not improve early postoperative outcomes. A prospective, randomized, controlled trial. AB - BACKGROUND: Minimally invasive total hip arthroplasty has stirred substantial controversy with regard to whether it provides superior outcomes compared with total hip arthroplasty performed through longer incisions. The orthopaedic literature is deficient in well-designed scientific studies to support the clinical superiority of this approach. The objective of this study was to compare the results of a single mini-incision approach with those of a standard-incision total hip arthroplasty in the early postoperative period. METHODS: Two hundred and nineteen patients (219 hips) admitted for unilateral total hip arthroplasty between December 2003 and June 2004 were randomized to undergo surgery through a short incision of 0.05). CONCLUSIONS: Although anteverting the humeral component during total shoulder arthroplasty to compensate for glenoid retroversion has been advocated, these data suggest that compensatory anteversion of the humeral component does not increase the stability of a shoulder replacement with a retroverted glenoid component. PMID- 15805212 TI - In vivo degradation of polyethylene liners after gamma sterilization in air. AB - BACKGROUND: Ultra-high molecular weight polyethylene degrades during storage in air following gamma sterilization, but the extent of in vivo degradation remains unclear. The purpose of this study was to quantify the extent to which the mechanical properties and oxidation of conventional polyethylene acetabular liners treated with gamma sterilization in air change in vivo. METHODS: Fourteen modular cementless acetabular liners were revised at an average of 10.3 years (range, 5.9 to 13.5 years) after implantation. All liners, which had been machined from GUR 415 resin, had been gamma-sterilized in air; the average shelf life was 0.3 year (range, 0.0 to 0.8 year). After removal, the components were expeditiously frozen to minimize ex vivo changes to the polyethylene prior to characterization. The average duration between freezing and testing was 0.6 year. Mechanical properties and oxidation were measured with use of the small-punch test and Fourier transform infrared spectroscopy, respectively, in the loaded and unloaded regions of the liners. RESULTS: There was substantial regional variation in the mechanical properties and oxidation of the retrieved liners. The ultimate load was observed to vary by >90% near the surface. On the average, the rim and the unloaded bearing showed evidence of severe oxidation near the surface after long-term in vivo aging, but these trends were not typically observed on the loaded bearing surface or near the backside of the liners. CONCLUSIONS: The mechanical properties of polyethylene that has been gamma-sterilized in air may decrease substantially in vivo, depending on the location in the liner. The most severe oxidation was observed at the rim, suggesting that the femoral head inhibits access of oxygen-containing body fluids to the bearing surface. This is perhaps why in vivo oxidation has not been associated with clinical performance to date. PMID- 15805213 TI - A cadaveric model of the throwing shoulder: a possible etiology of superior labrum anterior-to-posterior lesions. AB - BACKGROUND: It has been speculated that a shift of the throwing arc commonly develops in athletes who perform overhead activities, resulting in greater external rotation and decreased internal rotation caused by anterior capsular laxity and posterior capsular contracture, respectively. Osseous adaptation in the form of increased humeral and glenoid retroversion may provide a protective function in the asymptomatic athlete but cannot explain the pathological changes seen in the shoulder of the throwing athlete. Therefore, the objective of the present study was to examine the biomechanical effects of capsular changes in a cadaveric model. METHODS: Ten cadaveric shoulders were tested with a custom shoulder-testing device. Humeral rotational range of motion, the position of the humerus in maximum external rotation, and glenohumeral translations in the anterior, posterior, superior, and inferior directions were measured with the shoulder in 90 degrees of abduction. Translations were measured with the humerus secured in 90 degrees of external rotation. To simulate anterior laxity due to posterior capsular contracture, the capsule was nondestructively stretched 30% beyond maximum external rotation with the shoulder in 90 degrees of abduction. This was followed by the creation of a 10-mm posterior capsular contracture. Rotational, humeral shift, and translational tests were performed for the intact normal shoulder, after anterior capsular stretching, and after simulated posterior capsular contracture. RESULTS: Nondestructive capsular stretching resulted in a significant increase in external rotation (average increase, 18.2 degrees 2.1 degrees ; p < 0.001), and subsequent simulated posterior capsular contracture resulted in a significant decrease in internal rotation (average decrease, 8.8 degrees +/- 2.3 degrees ; p = 0.02). There also was a significant increase in anterior translation with the application of a 20-N anterior translational force after nondestructive capsular stretching (average increase, 1.7 +/- 0.3 mm, p = 0.0006). The humeral head translated posteroinferiorly when the humerus was rotated from neutral to maximum external rotation. This did not change significantly in association with anterior capsular stretching. Following simulated posterior capsular contracture, there was a trend toward a more posterosuperior position of the humeral head with the humerus in maximum external rotation in comparison with the position in the stretched conditions, although these differences were not significant. CONCLUSIONS: A posterior capsular contracture with decreased internal rotation does not allow the humerus to externally rotate into its normal posteroinferior position in the cocking phase of throwing. Instead, the humeral head is forced posterosuperiorly, which may explain the etiology of Type-II superior labrum anterior-to-posterior lesions in overhead athletes. PMID- 15805214 TI - Soft-tissue releases to treat spastic hip subluxation in children with cerebral palsy. AB - BACKGROUND: Spastic hip subluxation and dislocation are common problems in children with cerebral palsy. Soft-tissue releases have proved to be beneficial in the prevention of spastic hip dislocation. A protocol for treatment based on patient age, hip abduction, and hip migration percentage was established in 1988. The purpose of this study was to assess the outcome in sixty-five children treated according to this protocol and followed for a minimum of eight years. METHODS: The medical records and radiographs of sixty-five children, from an original series of seventy-four patients, who met the inclusion criteria were available for review. Forty-seven children had spastic quadriplegia and were unable to walk; eighteen children had spastic diplegia and were able to walk independently or with assistive devices. The mean age at the time of the surgery was 4.4 years. Open adductor tenotomy and psoas muscle recession or iliopsoas tenotomy were performed on 129 hips, which were followed for a mean of 10.8 years. The mean age at the time of follow-up was fifteen years. Hips were grouped according to the hip migration percentage preoperatively, at one year postoperatively, and at the time of final follow-up. The final outcome for the patient was defined according to the worse hip. An analysis was performed to identify potential factors influencing outcome. RESULTS: Thirty-two patients (49%) had a good result, eleven (17%) had a fair result, three (4%) had a poor result, and nineteen (30%) had a failure. The mean hip migration percentage was 34% preoperatively and 18% at the time of final follow-up. Nineteen patients required subsequent osseous reconstructive procedures, and eleven required repeat soft-tissue releases. The migration percentage at one year postoperatively was the most predictive of the final outcome (p = 0.001). Patients who had been able to walk preoperatively had a better long-term outcome (p = 0.01). Neither the preoperative hip migration percentage nor the age at surgery significantly affected the outcome. CONCLUSIONS: Soft-tissue release was effective for long term prevention of hip dislocation in 67% (forty-three) of sixty-five children with spastic hip subluxation. Two preoperative factors that were related to a favorable outcome were a spastic diplegic pattern of involvement and the ability to walk. The hip migration percentage at one year postoperatively was a good predictor of final outcome. PMID- 15805215 TI - Periprosthetic infection in patients treated for an orthopaedic oncological condition. AB - BACKGROUND: Prosthetic replacement following excision of a bone tumor can be complicated by infection because patients who undergo surgery for a neoplastic condition often are subjected to extensive soft-tissue dissection and long operating times and are immunosuppressed. The aim of this study was to investigate the rate of periprosthetic infection, identify possible predisposing factors, and assess treatment efficacy in such patients. METHODS: Prosthetic reconstruction was performed in 1264 patients over a thirty-seven-year period in a single hospital by four surgeons. Twenty-four patients were excluded because of incomplete follow-up, leaving 1240 patients who had been followed for a mean of 5.8 years. Infection was identified in 136 patients (11.0%). The management and outcome of the infections in all of these patients were analyzed. RESULTS: Coagulase-negative Staphylococcus was the most common organism isolated. Two stage revision successfully treated the infection in 72% (forty-two) of the fifty eight patients in whom it was performed. Local surgical debridement with or without antibiotic implants was successful in only 6% (four) of sixty-eight patients. Amputation to treat the infection was performed in fifty (37%) of the 136 patients. The factors that were associated with a significant risk of infection (p or=40% of the length of the bone). Another twenty age matched control subjects were recruited for this study. Knee proprioception was evaluated by comparing active and passive angular repositioning of the involved limb and the normal limb in the patients and the dominant limb in the central subjects. RESULTS: The absolute matching error of active and passive repositioning was 2.7 degrees +/- 2.1 degrees and 2.9 degrees +/- 2.2 degrees , respectively, for knees treated operatively and 3.2 degrees +/- 2.1 degrees and 2.1 degrees +/- 1.3 degrees , respectively, for the dominant knees in the control subjects. The differences between the operatively treated knees and the control knees were not significant (p > 0.05). There was also no significant difference in proprioception between the patients with a distal femoral tumor and those with a proximal tibial tumor (p > 0.05). The proprioception of the patients with a greater length of bone resected was significantly worse in both lower extremities than that of patients with a shorter resected length (p = 0.016 for the normal side and p = 0.025 for the involved side). CONCLUSIONS: The proprioception of the involved knees in the patients with osteosarcoma who had modular endoprosthetic reconstruction was not found to be different from the normal knees or the knees in the control subjects. On the basis of our data, we found that proprioception is probably not influenced by the tumor site. Conversely, resection length appears to be an important factor. These study findings may be used as a reference for knee proprioception in patients with osteosarcoma who have a modular endoprosthetic knee reconstruction. PMID- 15805217 TI - Arthroscopic removal of the glenoid component for failed total shoulder arthroplasty. A report of five cases. PMID- 15805218 TI - Total sacrectomy and reconstruction with structural allografts for neurofibrosarcoma of the sacrum. A case report. PMID- 15805219 TI - Persistent bladder entrapment following acetabular fracture with subsequent vesical injury during total hip arthroplasty. A case report. PMID- 15805220 TI - Spontaneous Clostridium septicum septic arthritis of the shoulder and gas gangrene. a case report. PMID- 15805221 TI - Lateral malleolar reconstruction after distal fibular resection. A case report. PMID- 15805222 TI - Recurrent posterior shoulder instability. AB - Recurrent posterior shoulder instability is an uncommon, debilitating condition in young adults that is being diagnosed with increasing frequency. Although a number of predisposing factors have been identified, their relative importance remains poorly understood. Poor results have been reported following operative intervention to treat recurrent posterior instability with nonanatomic techniques. The more recent development of lesion-specific surgery has improved clinical results, particularly when that surgery has been performed arthroscopically. Operative treatment is therefore being increasingly recommended at an earlier stage to patients who do not respond to supervised rehabilitation programs. PMID- 15805223 TI - The early effects of code 405 work rules on attitudes of orthopaedic residents and attending surgeons. AB - BACKGROUND: The impact of strict enforcement of Section 405 of the New York State Public Health Code to restrict resident work to eighty hours per week and the adoption of a similar policy by the Accreditation Council on Graduate Medical Education in 2002 for orthopaedic residency training have not been evaluated. Adoption of these rules has created accreditation as well as staffing problems and has generated controversy in the surgical training community. The purposes of this study were (1) to evaluate the attitudes of orthopaedic residents and attending surgeons toward the Code 405 work-hour regulations and the effect of those regulations on the perceived quality of residency training, quality of life, and patient care and (2) to quantify the effect of the work-hour restrictions on the actual number of hours worked. METHODS: We administered a thirty-four-question Likert-style questionnaire to forty-eight orthopaedic surgery residents (postgraduate years [PGY]-2 through 5) and a similar twenty nine-question Likert-style questionnaire to thirty-nine orthopaedic attending surgeons. All questionnaires were collected anonymously and analyzed. Additionally, resident work hours before and after strict enforcement of the Code 405 regulations were obtained from resident time sheets. RESULTS: The average weekly work hours decreased from 89.25 to 74.25 hours for PGY-2 residents and from 86.5 to 73.25 hours for PGY-3 residents, and they increased from 61.5 to 68.5 hours for PGY-4 residents. Residents at all levels felt that they had increased time available for reading. There was general agreement between attending and resident surgeons that their operating experience had been negatively impacted. Senior residents thought that their education had been negatively affected, while junior residents thought that their operating experience in general had been negatively affected. Senior residents and attending surgeons felt that continuity of care had been negatively impacted. All agreed that quality of life for the residents had improved and that residents were more rested. CONCLUSIONS: On the basis of the survey data, the implementation of the new work-hour restrictions was found to result in a decrease in the number of hours worked per week for PGY-2 and PGY-3 residents and in an increase in work hours for PGY-4 residents. This could explain the definite difference between the attitudes expressed by the senior residents and those of the junior residents. Senior residents felt that their education was negatively impacted by the work rules, while junior residents expressed a more neutral view. However, senior residents did not believe that their operative experience was as negatively impacted as did junior residents. Although junior and senior residents and attending surgeons agreed that resident quality of life had improved, we were not able to determine whether this offset the perceived negative impact on education, continuity of care, and operative experience. PMID- 15805224 TI - What's new in foot and ankle surgery? PMID- 15805225 TI - The RXR-type endoplasmic reticulum-retention/retrieval signal of GABAB1 requires distant spacing from the membrane to function. AB - Functional gamma-aminobutyric acid type B (GABA(B)) receptors are normally only observed upon coexpression of GABA(B1) with GABA(B2) subunits. A C-terminal arginine-based endoplasmic reticulum (ER) retention/retrieval signal, RSRR, prevents escape of unassembled GABA(B1) subunits from the ER and restricts surface expression to correctly assembled heteromeric receptors. The RSRR signal in GABA(B1) is proposed to be shielded by C-terminal coiled-coil interaction of the GABA(B1) with the GABA(B2) subunit. Here, we investigated whether the RSRR motif in GABA(B1) remains functional when grafted to ectopic sites. We found that the RSRR signal in GABA(B1) is inactive in any of the three intracellular loops but remains functional when moved within the distal zone of the C-terminal tail. C-terminal deletions that position the RSRR signal closer to the plasma membrane drastically reduce its effectiveness, supporting that proximity to the membrane restricts access to the RSRR motif. Functional ectopic RSRR signals in GABA(B1) are efficiently inactivated by the GABA(B2) subunit in the absence of coiled-coil dimerization, supporting that coiled-coil interaction is not critical for release of the receptor complex from the ER. The data are consistent with a model in which removal of RSRR from its active zone rather than its direct shielding by coiled-coil dimerization triggers forward trafficking. Because arginine-based intracellular retention signals of the type RXR, where X represents any amino acid, are used to regulate assembly and surface transport of several multimeric complexes, such a mechanism may apply to other proteins as well. PMID- 15805226 TI - Blue flicker modifies the subfoveal choroidal blood flow in the human eye. AB - The objective of the present study was to reveal an interaction between choroidal blood flow (ChBF) and light-induced photoreceptor activity, a physiological coupling that has been already demonstrated for retinal blood flow but rejected for ChBF. Ten healthy adults volunteered for this study. A real-time recording near-infrared laser-Doppler flowmeter was used to quantify the subfoveal ChBF while the luminance of blue flicker between 1 and 64 Hz was first increased and then decreased by 4.0 log units in 1.0-log unit steps between 0.0375 and 375 cd/m2. In separate testing, flash electroretinograms (ERGs) provided electrophysiological indexes of the relative response of short-wave cones (s cones) and rods to blue light stimulation. Group-averaged, normalized ChBF measurements revealed that it was modulated by approximately 9% by flicker frequency. Increasing the blue flicker luminance from low to high attenuated the subfoveal ChBF, volume, and velocity by approximately 32%, approximately 30%, and approximately 5%, respectively. Decreasing the luminance from high to low over the same range had no effect on the subfoveal choroidal hemodynamics. The markedly different effects of reversed directions of change in blue flicker luminance on the subfoveal ChBF were linked to transitions between rod-dominated and s-cone-dominated retinal responses. Collectively, these findings indicate that the blue light-induced photoreceptor response is associated with a differential distribution of the ChBF across the ocular fundus according to the degree and type of retinal photoreceptor stimulated. PMID- 15805227 TI - Paracrine mitogenic effect of human endothelial progenitor cells: role of interleukin-8. AB - Endothelial progenitor cells (EPCs) play an important role in repair of vascular injury and neovascularization. Molecular mechanisms underlying vascular effects of EPCs are not fully understood. The present study was designed to test the hypothesis that human EPCs exert a strong paracrine mitogenic effect on mature endothelial cells. Levels of interleukin-8 (IL-8) were significantly higher in conditioned medium (CM) collected from EPCs than in CM derived from mature endothelial cells [umbilical vein endothelial cells (HUVECs) and coronary artery endothelial cells (CAECs)]. CM of EPCs stimulated proliferation of HUVECs and CAECs. This mitogenic effect was partially inhibited by IL-8-neutralizing antibody. In contrast, CM of HUVECs and CAECs had a weak or no mitogenic effect on mature endothelial cells. Our results demonstrate significantly higher levels of IL-8 secretion by human EPCs than by mature endothelial cells. IL-8 appears to be an important mediator of the paracrine mitogenic effect of EPCs. PMID- 15805228 TI - Revealing anti-inflammatory mechanisms of soy isoflavones by flow: modulation of leukocyte-endothelial cell interactions. AB - The antiatherogenic effects of soy isoflavone consumption have been demonstrated in a variety of studies. However, the mechanisms involved remain poorly defined. Adhesion of monocytes to vascular endothelial cells is a key step within the inflammatory cascade that leads to atherogenesis. Many factors, including the physical forces associated with blood flow, regulate this process. Using an in vitro flow assay, we report that genistein, a principal component of most isoflavone preparations, inhibits monocyte adhesion to cytokine (TNF-alpha) stimulated human vascular endothelial cells at physiologically relevant concentrations (0-1 microM). This effect is absolutely dependent on flow and is not observed under static conditions. Furthermore, this inhibition was dependent on activation of endothelial peroxisomal proliferator-activated receptor-gamma. No significant role for other reported properties of genistein, including antioxidant effects, inhibition of tyrosine kinases, or activation of estrogen receptors, was observed. Furthermore, the antiadhesive effects of genistein did not occur via modulation of the adhesion molecules E-selectin, ICAM-1, VCAM-1, or platelet-endothelial cell adhesion molecule-1. These data reveal a novel anti inflammatory mechanism for isoflavones and identify the physical forces associated with blood flow and a critical mediator of this function. PMID- 15805229 TI - Influence of insulin and free fatty acids on contractile function in patients with chronically stunned and hibernating myocardium. AB - It is unknown whether short-term modulation of substrate supply affects cardiac performance in heart failure patients with chronic ischemic myocardium. The aim of this study was to determine whether modulation of myocardial substrate metabolism with insulin and free fatty acids (FFAs) affects contractile function of chronically stunned (CST) and hibernating (HIB) myocardium at rest and after maximal exercise. We studied eight nondiabetic patients with ejection fraction (EF) 30 +/- 4% (SE) and CST/HIB in 49 +/- 6% of the left ventricle: 36 +/- 6% CST and 13 +/- 2% HIB as determined by 99m Technetium-Sestamibi single photon emission computed tomography (SPECT) and [18F]fluorodeoxyglucose (FDG) positron emission tomography (PET). Each patient was subjected to a 3-h infusion of 1) saline, 2) insulin-glucose (i.e., euglycemic insulin clamp; high insulin, suppressed FFA), and 3) somatostatin-heparin (suppressed insulin, high FFA). Echocardiographic endpoints were global EF and regional contractile function [maximum velocity (Vmax) and strain rate (epsilon max)] as determined by tissue Doppler imaging at steady state and after maximal exercise. EF was similar at baseline and steady state and increased after exercise to 36 +/- 5% (P < 0.05). Baseline regional Vmax and epsilon max were highest in control, intermediate in CST and HIB, and lowest in infarct regions (P < 0.05). Steady-state EF, Vmax, and epsilon max were not affected by metabolic modulation in any region. After maximal exercise, contractile function increased in control, CST, and HIB (P < 0.05), but not in infarct, regions. Exercise-induced contractile increments were unaffected by metabolic modulation. Metabolic modulation does not influence contractile function in CST and HIB regions. Chronic ischemic myocardium has preserved ability to adapt to extreme, short-term changes in substrate supply at rest and after maximal exercise. PMID- 15805230 TI - HIF-1 activation attenuates postischemic myocardial injury: role for heme oxygenase-1 in modulating microvascular chemokine generation. AB - The CXC chemokine IL-8, which promotes adhesion, activation, and transmigration of polymorphonuclear neutrophils (PMN), has been associated with production of tissue injury in reperfused myocardium. Hypoxia-inducible factor-1 (HIF-1) is a heterodimeric peptide that is a key regulator of genes such as heme oxygenase (HO)-1 expressed under hypoxic conditions. We hypothesized that HO-1 plays an important role in regulating proinflammatory mediator production under conditions of ischemia-reperfusion. HIF-1 was activated in the human microvascular endothelial cell line (HMEC-1) with the prolyl hydroxylase inhibitor dimethyloxalylglycine (DMOG). DMOG significantly attenuated cytokine-induced IL-8 promoter activity and protein secretion and cytokine-induced PMN migration across human microvascular endothelial cell line HMEC-1 monolayers. In vivo studies in a rabbit model of myocardial ischemia-reperfusion showed that rabbits pretreated with a 20 mg/kg DMOG infusion (n = 6) 24 h before study exhibited a 21.58 +/- 1.76% infarct size compared with 35.25 +/- 2.06% in saline-treated ischemia reperfusion animals (n = 6, change in reduction = 39%; P < 0.001). In DMOG pretreated (20 mg/kg) animals, plasma IL-8 levels at 3 h after onset of reperfusion were 405 +/- 40 pg/ml vs. 790 +/- 40 pg/ml in saline-treated ischemia reperfusion animals (P < 0.001). DMOG pretreatment reduced myocardial myeloperoxidase activity, expressed as number of PMN per gram of myocardium, to 1.43 +/- 0.59 vs. 4.86 +/- 1.1 (P = 0.012) in saline-treated ischemia-reperfused hearts. Both in vitro and in vivo DMOG-attenuated IL-8 production was associated with robust HO-1 expression. Thus our data show that HIF-1 activation induces substantial HO-1 expression that is associated with attenuated proinflammatory chemokine production by microvascular endothelium in vitro and in vivo. PMID- 15805231 TI - Cardiomyocyte apoptosis induced by short-term diabetes requires mitochondrial GSH depletion. AB - Oxidative stress due to excessive reactive oxygen species (ROS) and depleted antioxidants such as glutathione (GSH) can give rise to apoptotic cell death in acutely diabetic hearts and lead to heart disease. At present, the source of these cardiac ROS or the subcellular site of cardiac GSH loss [i.e., cytosolic (cGSH) or mitochondrial (mGSH) GSH] has not been completely elucidated. With the use of rotenone (an inhibitor of the electron transport chain) to decrease the excessive ROS in acute streptozotocin (STZ)-induced diabetic rat heart, the mitochondrial origin of ROS was established. Furthermore, mitochondrial damage, as evidenced by loss of membrane potential, increases in oxidative stress, and reduction in mGSH was associated with increased apoptosis via increases in caspase-9 and -3 activities in acutely diabetic hearts. To validate the role of mGSH in regulating cardiac apoptosis, L-buthionine-sulfoximine (BSO; 10 mmol/kg ip), which blocks GSH synthesis, or diethyl maleate (DEM; 4 mmol/kg ip), which inactivates preformed GSH, was administered in diabetic rats for 4 days after STZ administration. Although both BSO and DEM lowered cGSH, they were ineffective in reducing mGSH or augmenting cardiomyocyte apoptosis. To circumvent the lack of mGSH depletion, BSO and DEM were coadministered in diabetic rats. In this setting, mGSH was undetectable and cardiac apoptosis was further aggravated compared with the untreated diabetic group. In a separate group, GSH supplementation induced a robust amplification of mGSH in diabetic rat hearts and prevented apoptosis. Our data suggest for the first time that mGSH is crucial for modulating the cell suicide program in short-term diabetic rat hearts. PMID- 15805232 TI - Oxidized LDL induces mitochondrially associated reactive oxygen/nitrogen species formation in endothelial cells. AB - Exposure of cells to complex mixtures of oxidized lipids such as those found in oxidized low-density lipoprotein (oxLDL) induce reactive oxygen and nitrogen species (ROS/RNS) formation. The source of the ROS/RNS within cells is unknown; it is thought they may be involved in redox cell signaling. Although this possibility was initially overlooked, it is becoming clear that mitochondria, which are a source of superoxide and hydrogen peroxide, may play a critical role in the response of cells on exposure to oxidized lipids. In this study, we tested the possibility that mitochondria are a potential source of oxLDL-dependent formation of ROS/RNS in endothelial cells. Using confocal microscopy, we demonstrated that a significant proportion of oxLDL-dependent dichlorodihydrofluorescein (DCF) fluorescence is colocalized to mitochondria. In support of this concept, rho0 endothelial cells showed a substantial decrease in ROS/RNS formation stimulated by oxLDL. In contrast, mostly nonmitochondrial DCF fluorescence was detected in cells exposed to an extracellular source of hydrogen peroxide. The exposure of cells to a nitric oxide synthase inhibitor and urate resulted in a decrease in oxLDL-induced DCF fluorescence that was restored by addition of nitric oxide donors to the medium. Taken together, these results suggest that oxLDL-dependent DCF fluorescence is mitochondrially associated and may be due to the formation of peroxynitrite. PMID- 15805234 TI - Orthostatic edema in adolescents: more than walking on water. PMID- 15805233 TI - Integrated pharmacological preconditioning and memory of cardioprotection: role of protein kinase C and phosphatidylinositol 3-kinase. AB - Although protein kinase C (PKC) and phosphatidylinositol 3 (PI3)-kinase are implicated in cardioprotective signal transduction mediated by ischemic preconditioning, their role in pharmacological preconditioning (PPC) has not been determined. Cultured neonatal rat cardiomyocytes (CMCs) were subjected to simulated ischemia for 2 h followed by 15 min of reoxygenation. PPC of CMCs consisted of administration of 50 microM adenosine, 50 microM diazoxide, and 50 microM S-nitroso-N-acetylpenicillamine (SNAP), each alone or in combination, for 15 min followed by 30 min of washout before simulated ischemia. Although PKC epsilon and PI3-kinase were significantly activated during treatment with adenosine, activation of these kinases dissipated after washout. In contrast, PPC combined with adenosine, diazoxide, and SNAP elicited sustained activation of PKC epsilon and PI-3 kinase after washout. The combined-PPC, but not the single-PPC, protocol conferred antiapoptotic and antinecrotic effects after reoxygenation. The PKC inhibitor chelerythrine (5 microM) or the PI3-kinase inhibitor LY-294002 (10 microM) given during the washout period partially blocked the activation of PKC-epsilon and PI3-kinase mediated by the combined-PPC protocol, whereas combined addition of chelerythrine and LY-294002 completely inhibited activation of PKC-epsilon and PI3-kinase. Chelerythrine or LY-294002 partially blocked antiapoptotic and antinecrotic effects mediated by the combined-PPC protocol, whereas combined addition of chelerythrine and LY-294002 completely abrogated antiapoptotic and antinecrotic effects. These results suggest that the combined PPC protocol confers cardioprotective memory through sustained and interdependent activation of PKC and PI3-kinase. PMID- 15805235 TI - Rocky Mountain spotted fever: a physician's challenge. PMID- 15805236 TI - Evaluation and management of language and speech disorders in preschool children. PMID- 15805237 TI - Index of suspicion. PMID- 15805238 TI - Treating hypernatremic dehydration. PMID- 15805239 TI - A new perspective on tumor endothelial cells: unexpected chromosome and centrosome abnormalities. AB - It has been shown that endothelial cells in solid tumors are cytogenetically abnormal. These cells are aneuploid with multiple chromosomes and multiple centrosomes. Unlike normal endothelial cells which remain diploid in long-term culture, the aneuploidy of tumor endothelial cells is exacerbated in culture suggesting that these cells are inherently unstable. It is speculated that this instability might compromise the effectiveness of antiangiogenesis therapy. PMID- 15805240 TI - Signaling intricacies take center stage in cancer cells. AB - After many years of productive study on the signaling networks, posttranslational regulatory control of effector molecules remains an intensely investigated and continuously evolving field of research to connect signaling with phenotypic changes. In recent years, there have been intriguing results on the interaction of critical molecules to control the growth of cancer cells. This review article will focus on two critical convergence signaling nodules, Akt and p21-activated kinase, two integral components of phenotypic signaling during tumorigenesis. Here we will summarize the recent findings on how these master signaling nodules regulate their targets and alter the subcellular localization of their effectors to control their functionality. Based on the laboratory advances in the Akt and p21-activated kinase signaling pathways, it is conceivable to start defining novel avenues to develop targeted anticancer therapies. PMID- 15805241 TI - Emerging role of RAB GTPases in cancer and human disease. AB - Emerging evidence implicates alterations in the RAB small GTPases and their associated regulatory proteins and effectors in multiple human diseases including cancer. We have recently shown that RAB25, located at chromosome 1q22, is amplified at the DNA level and overexpressed at the RNA level in ovarian and breast cancer. These changes correlated with a worsened outcome in both diseases. In addition, enforced expression of RAB25 in both breast and ovarian cancer cells decreased apoptosis and increased proliferation and aggressiveness in vivo, potentially explaining the worsened prognosis. A better understanding of genetic alterations as well as the physiologic and pathophysiologic roles of RAB GTPases may open new opportunities for therapeutic intervention and better outcomes. PMID- 15805242 TI - Tid-1 interacts with the von Hippel-Lindau protein and modulates angiogenesis by destabilization of HIF-1alpha. AB - The von Hippel-Lindau protein (pVHL) is a major tumor suppressor protein and also associated with the inhibition of angiogenesis via HIF-1alpha ubiquitination and proteasomal degradation. To further elucidate the biological activity of pVHL in angiogenesis, pVHL-interacting proteins were screened using the yeast two-hybrid system. We found that a mouse homologue of the long form of Drosophila tumor suppressor l(2)tid, Tid-1(L), directly interacts with pVHL in vitro and in vivo. Furthermore, Tid-1(L) protein; enhanced the interaction between HIF-1alpha and pVHL, leading to the destabilization of HIF-1alpha protein; therefore, Tid-1(L) protein decreased vascular endothelial growth factor expression and inhibited angiogenesis in vivo and in vitro. These findings propose that Tid-1(L) may play a critical role in pVHL-mediated tumor suppression by modulating the pVHL dependent HIF-1alpha stability. PMID- 15805243 TI - BLM helicase facilitates Mus81 endonuclease activity in human cells. AB - Bloom syndrome is a rare, autosomal recessive inherited disorder in humans. The product of the Bloom syndrome mutated gene, designated BLM, is a member of the RecQ helicase family. BLM has been proposed to function at the interface of replication and recombination, and to facilitate the repair of DNA damage. Here, we report in vivo physical interaction and colocalization of BLM and a DNA structure-specific endonuclease, Mus81, at sites of stalled replication forks outside the promyelocytic leukemia nuclear bodies during the S-phase arrest of the cell cycle. Amino acids 125 to 244 of Mus81 interact with the C-terminal region (amino acids 1,007-1,417) of BLM. Whereas Mus81 does not have any effect on the helicase activity of BLM, BLM can stimulate Mus81 endonuclease activity on the nicked Holliday junctions and 3' flap. This stimulation is due to enhanced binding of Mus81 to the DNA substrates. These data suggest a new function of BLM in cooperating with Mus81 during processing and restoration of stalled replication forks. PMID- 15805244 TI - Knockdown of STAT3 expression by RNA interference inhibits the induction of breast tumors in immunocompetent mice. AB - Constitutively activated STAT3 is involved in the formation of multiple types of tumors including breast cancer. We examined the effects of Stat3 protein knockdown by RNA interference using a dicistronic lentivirus small hairpin (shRNA) delivery system on the growth of mammary tumors in BALB/c mice induced by the 4T1 cell line. A single exposure of 4T1 cells to shRNA/STAT3 lentivirus transduced 75% of the cells with green fluorescent protein (GFP) within 96 hours. In cells selected for GFP expression, neither Stat3 protein nor phosphotyrosine Stat3 was detected. Tumor formation induced by injecting 4T1 cells into the mammary fat pad was blocked by expression of the shRNA for STAT3 whereas all mice injected with 4T1 cells expressing only GFP efficiently formed tumors. c-Myc expression was reduced 75% in cells expressing greatly reduced levels of Stat3 compared with the GFP control. Of interest, the level of activated Src, which is known to activate Stat3, was virtually eliminated but the level of the Src protein itself remained the same. Importantly, expression of Twist protein, a metastatic regulator, was eliminated in STAT3 knockdown cells. Invasion activity of STAT3 knockdown cells was strongly inhibited. However, the proliferation rate of cells in Stat3 knockdown cells was similar to that of the GFP control; the cell cycle was also not affected. We conclude from these studies that activated Stat3 protein plays a critical role in the induction of breast tumors induced by 4T1 cells by enhancing the expression of several important genes including c-Myc and the metastatic regulator Twist. These studies suggest that stable expression of small interfering RNA for STAT3 has potential as a therapeutic strategy for breast cancer. PMID- 15805245 TI - Valproic acid stimulates proliferation and self-renewal of hematopoietic stem cells. AB - Histone deacetylase inhibitors have attracted considerable attention because of their ability to overcome the differentiation block in leukemic blasts, an effect achieved either alone or in combination with differentiating agents, such as all trans retinoic acid. We have previously reported favorable effects of the potent histone deacetylase inhibitor valproic acid in combination with all-trans retinoic acid in patients with advanced acute myeloid leukemia leading to blast cell reduction and improvement of hemoglobin. These effects were accompanied by hypergranulocytosis most likely due to an enhancement of nonleukemic myelopoiesis and the suppression of malignant hematopoiesis rather than enforced differentiation of the leukemic cells. These data prompted us to investigate the effect of valproic acid on normal hematopoietic stem cells (HSC). Here we show that valproic acid increases both proliferation and self-renewal of HSC. It accelerates cell cycle progression of HSC accompanied by a down-regulation of p21(cip-1/waf-1). Furthermore, valproic acid inhibits GSK3beta by phosphorylation on Ser9 accompanied by an activation of the Wnt signaling pathway as well as by an up-regulation of HoxB4, a target gene of Wnt signaling. Both are known to directly stimulate the proliferation of HSC and to expand the HSC pool. In summary, we here show that valproic acid, known to induce differentiation or apoptosis in leukemic blasts, stimulates the proliferation of normal HSC, an effect with a potential effect on its future role in the treatment of acute myeloid leukemia. PMID- 15805246 TI - Genome-wide association study in esophageal cancer using GeneChip mapping 10K array. AB - Whole genome association studies of complex human diseases represent a new paradigm in the postgenomic era. In this study, we report application of the Affymetrix, Inc. (Santa Clara, CA) high-density single nucleotide polymorphism (SNP) array containing 11,555 SNPs in a pilot case-control study of esophageal squamous cell carcinoma (ESCC) that included the analysis of germ line samples from 50 ESCC patients and 50 matched controls. The average genotyping call rate for the 100 samples analyzed was 96%. Using the generalized linear model (GLM) with adjustment for potential confounders and multiple comparisons, we identified 37 SNPs associated with disease, assuming a recessive mode of transmission; similarly, 48 SNPs were identified assuming a dominant mode and 53 SNPs in a continuous mode. When the 37 SNPs identified from the GLM recessive mode were used in a principal components analysis, the first principal component correctly predicted 46 of 50 cases and 47 of 50 controls. Among all the SNPs selected from GLMs for the three modes of transmission, 39 could be mapped to 1 of 33 genes. Many of these genes are involved in various cancers, including GASC1, shown previously to be amplified in ESCCs, and EPHB1 and PIK3C3. In conclusion, we have shown the feasibility of the Affymetrix 10K SNP array in genome-wide association studies of common cancers and identified new candidate loci to study in ESCC. PMID- 15805247 TI - Post-transcriptional regulation of the androgen receptor by Mammalian target of rapamycin. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF), an ErbB1 ligand and prostate stromal growth factor, is an antagonist of androgen receptor (AR) function. In the LNCaP prostate cancer model, HB-EGF reduced AR protein levels and AR transactivation without affecting AR mRNA level or protein turnover. The signal to attenuate AR was mediated by the mammalian target of rapamycin, as shown by genetic and pharmacologic methods, and was independent of ErbB2/HER-2, extracellular signal-regulated kinase 1/2, and p38 mitogen-activated protein kinase pathways. Additional evidence suggests that AR protein levels are highly sensitive to regulation by cap-dependent mRNA translation. These findings reveal a novel mechanism for regulation of AR by a classic growth factor system and indicate that a rapamycin-sensitive post-transcriptional pathway can attenuate or possibly bypass AR-mediated signaling. PMID- 15805248 TI - PIK3CA mutations correlate with hormone receptors, node metastasis, and ERBB2, and are mutually exclusive with PTEN loss in human breast carcinoma. AB - Deregulation of the phosphatidylinositol 3-kinase (PI3K) pathway either through loss of PTEN or mutation of the catalytic subunit alpha of PI3K (PIK3CA) occurs frequently in human cancer. We identified PIK3CA mutations in 26% of 342 human breast tumor samples and cell lines at about equal frequency in tumor stages I to IV. To investigate the relationship between PTEN and PIK3CA, we generated a cohort of tumors that had lost PTEN expression and compared it with a matched control set that had retained PTEN. A highly significant association between PIK3CA mutations and retention of PTEN protein expression was observed. In addition, PIK3CA mutations were associated with expression of estrogen and progesterone receptors (ER/PR), lymph node metastasis, and ERBB2 overexpression. The fact that PIK3CA mutations and PTEN loss are nearly mutually exclusive implies that deregulated phosphatidylinositol-3,4,5-triphosphate (PIP(3)) is critical for tumorigenesis in a significant fraction of breast cancers and that loss of PIP(3) homeostasis by abrogation of either PIK3CA or PTEN relieves selective pressure for targeting of the other gene. The correlation of PIK3CA mutation to ER/PR-positive tumors and PTEN loss to ER/PR-negative tumors argues for disparate branches of tumor evolution. Furthermore, the association between ERBB2 overexpression and PIK3CA mutation implies that more than one input activating the PI3K/AKT pathway may be required to overcome intact PTEN. Thus, mutation of PIK3CA is frequent, occurs early in carcinoma development, and has prognostic and therapeutic implications. PMID- 15805249 TI - CY15, a malignant histiocytic tumor that is phenotypically similar to immature dendritic cells. AB - The origin and pathogenesis of histiocytic malignancies and the biology of the tumor cells are poorly understood. We have isolated a murine histiocytic tumor cell line (CY15) from a BALB/c IFNgamma(-/-) mouse and characterized it in terms of phenotype and function. The morphology, as judged by electron microscopy, and the surface marker phenotype suggests that CY15 cells are similar to immature dendritic cells (CD11c (low), MHC II (low), CD11b(+), B7.1(+), B7.2(+), and CD40(+)). The cells form tumors in BALB/c mice and metastasize to spleen, liver, lung, kidney, and to a lesser extend to lymph nodes and bone marrow, as judged by the growth of green fluorescent protein transfected tumor cells in mice. CY15 cells are capable of actively taking up antigen (FITC-ovalbumin) and can stimulate T lymphocytes in an allogenic mixed lymphocyte reaction but less effectively than their normal counterparts (immature dendritic cells). They respond to interleukin 4 (IL-4) with up-regulation of CD11c. If stimulated with IFNgamma the cells up-regulate MHC II, CD40 B7.1, and B7.2. Lipopolysaccharide induces the cells to up-regulate B7.1 and B7.2 and to secrete tumor necrosis factor alpha and IL-12. Based on these data, CY15 is a dendritic cell-like tumor cell line and may serve as a transplantable tumor model for histiocytosis in humans. PMID- 15805250 TI - EMP3, a myelin-related gene located in the critical 19q13.3 region, is epigenetically silenced and exhibits features of a candidate tumor suppressor in glioma and neuroblastoma. AB - The presence of common genomic deletions in the 19q13 chromosomal region in neuroblastomas and gliomas strongly suggests the presence of a putative tumor suppressor gene for these neoplasms in this region that, despite much effort, has not yet been identified. In an attempt to address this issue, we compared the expression profile of 89 neuroblastoma tumors with that of benign ganglioneuromas by microarray analysis. Probe sets (637 of 62,839) were significantly down regulated in neuroblastoma tumors, including, most importantly, a gene located at 19q13.3: the epithelial membrane protein 3 (EMP3), a myelin-related gene involved in cell proliferation and cell-cell interactions. We found that EMP3 undergoes hypermethylation-mediated transcriptional silencing in neuroblastoma and glioma cancer cell lines, whereas the use of the demethylating agent 5-aza-2 deoxycytidine restores EMP3 gene expression. Furthermore, the reintroduction of EMP3 into neuroblastoma cell lines displaying methylation-dependent silencing of EMP3 induces tumor suppressor-like features, such as reduced colony formation density and tumor growth in nude mouse xenograft models. Screening a large collection of human primary neuroblastomas (n = 116) and gliomas (n = 41), we observed that EMP3 CpG island hypermethylation was present in 24% and 39% of these tumor types, respectively. Furthermore, the detection of EMP3 hypermethylation in neuroblastoma could be clinically relevant because it was associated with poor survival after the first 2 years of onset of the disease (Kaplan-Meier; P = 0.03) and death of disease (Kendall tau, P = 0.03; r = 0.19). Thus, EMP3 is a good candidate for being the long-sought tumor suppressor gene located at 19q13 in gliomas and neuroblastomas. PMID- 15805251 TI - Proximity of TPR and NTRK1 rearranging loci in human thyrocytes. AB - Chromosomal rearrangements are frequently associated with cancer; the mechanisms underlying their cell-type specificity are poorly understood. Papillary thyroid carcinomas are marked by a high frequency of chromosome rearrangements involving the RET and NTRK1 tyrosine kinase receptor genes and producing RET and TRK oncogenes. An explanation for the propensity of thyrocytes to undergo gene rearrangements has been recently proposed by Nikiforova and colleagues, who showed that the recombination between RET and H4 is favored by the loci proximity in interphase nuclei. We investigated whether the spatial proximity is a contributing factor also in the generation of the thyroid-specific TRK oncogenes. The distance between NTRK1 and its oncogenic partner TPR was determined by two color fluorescence in situ hybridization and two-dimensional microscopy. A three dimensional reconstruction of the data was also done. We show that the two loci in thyrocytes nuclei display a distance reduced with respect to peripheral blood lymphocytes, thus supporting the notion that spatial proximity of translocation prone gene loci may favor gene rearrangements. PMID- 15805252 TI - The effect of Bcrp1 (Abcg2) on the in vivo pharmacokinetics and brain penetration of imatinib mesylate (Gleevec): implications for the use of breast cancer resistance protein and P-glycoprotein inhibitors to enable the brain penetration of imatinib in patients. AB - Imatinib mesylate (signal transduction inhibitor 571, Gleevec) is a potent and selective tyrosine kinase inhibitor, which was shown to effectively inhibit platelet-derived growth factor-induced glioblastoma cell growth preclinically. However, in patients, a limited penetration of imatinib into the brain has been reported. Imatinib is transported in vitro and in vivo by P-glycoprotein (P-gp; ABCB1), which thereby limits its distribution into the brain in mice. Previously, imatinib was shown to potently inhibit human breast cancer resistance protein (BCRP; ABCG2). Here, we show that imatinib is efficiently transported by mouse Bcrp1 in transfected Madin-Darby canine kidney strain II (MDCKII) monolayers. Furthermore, we show that the clearance of i.v. imatinib is significantly decreased 1.6-fold in Bcrp1 knockout mice compared with wild-type mice. At t = 2 hours, the brain penetration of i.v. imatinib was significantly 2.5-fold increased in Bcrp1 knockout mice compared with control mice. We tested the hypothesis that P-gp and BCRP inhibitors, such as elacridar and pantoprazole, improve the brain penetration of imatinib. Firstly, we showed in vitro that pantoprazole and elacridar inhibit the Bcrp1-mediated transport of imatinib in MDCKII-Bcrp1 cells. Secondly, we showed that co-administration of pantoprazole or elacridar significantly reduced the clearance of i.v. imatinib in wild-type mice by respectively 1.7-fold and 1.5-fold. Finally, in wild-type mice treated with pantoprazole or elacridar, the brain penetration of i.v. imatinib significantly increased 1.8-fold and 4.2-fold, respectively. Moreover, the brain penetration of p.o. imatinib increased 5.2-fold when pantoprazole was co-administered in wild type mice. Our results suggest that co-administration of BCRP and P-gp inhibitors may improve delivery of imatinib to malignant gliomas. PMID- 15805253 TI - p53 mutations in benzo(a)pyrene-exposed human p53 knock-in murine fibroblasts correlate with p53 mutations in human lung tumors. AB - Human p53 mutation spectra differ significantly from one cancer type to another. One possible reason is that carcinogenic risk factors differ, and these factors elicit distinct mutation patterns. There has been no mammalian assay, however, with which to generate mutation patterns in human p53 sequences experimentally, hampering interpretation of the human tumor spectra. We have designed a new mammalian cell assay using gene targeting technology that selects and scores human p53 gene sequence mutations in human-p53 knock-in (Hupki) murine embryonic fibroblasts (HUF) that have undergone immortalization. With the Hupki assay we examined here whether benzo(a)pyrene (BaP), a major tobacco smoke carcinogen could elicit p53 mutation patterns characterizing the human lung tumor p53 mutation spectrum. We found that, in contrast to unexposed HUFs or HUFs exposed to other carcinogenic agents, HUFs exposed to BaP acquire mutations that display major features of the human lung tumor p53 mutation spectrum: (a) predominance of G-to-T mutations, (b) unequivocal strand bias of the transversions, and (c) a mutation hotspot at codons 157 to 158. These data are consistent with the hypothesis that BaP has a direct role in causing smokers' lung tumor p53 mutations. The assay can be used to examine various hypotheses on the endogenous or exogenous factors responsible for the p53 mutations in human tumors arising in other tissues. PMID- 15805254 TI - Endogenous retrovirus expression is required for murine melanoma tumor growth in vivo. AB - Tumor development is a multistep process in which both genetic and epigenetic events cooperate for the emergence of a malignant clone. The possibility that endogenous retroviruses promote the expansion of a neoplastic clone by subverting immune surveillance has been proposed, but remained elusive. Here we show that knocking down-by RNA interference-an endogenous retrovirus spontaneously induced in the B16 murine melanoma results in the rejection of the tumor cells in immunocompetent mice, under conditions where control melanoma cells grow into lethal tumors. The knockdown does not modify the transformed phenotype of the cells, as measured both in vitro by a soft agar assay and in vivo by tumor cell proliferation in immunoincompetent (X-irradiated and severe combined immunodeficiency) mice. Tumor rejection can be reverted upon adoptive transfer of regulatory T cells from control melanoma-engrafted mice, as well as upon reexpression of the sole envelope gene of the endogenous retrovirus in the knocked down cells. These results show that endogenous retroviruses can be essential for a regulatory T-cell-mediated subversion of immune surveillance and could be relevant to human tumors where such elements-and especially their envelope gene-are induced. PMID- 15805255 TI - A gene expression signature for relapse of primary wilms tumors. AB - Anaplastic histology and metastasis are each associated with higher relapse and mortality rates in Wilms tumor patients. However, not all anaplastic tumors relapse and some nonanaplastic tumors relapse unexpectedly. To identify more accurate early prognostic indicators, we analyzed expression of 4,900 cancer related genes in 26 primary Wilms tumors. This analysis revealed that expression of a set of four genes predicts future relapse of primary Wilms tumors with high accuracy, independent of anaplasia. Random permutation testing of this prognostic gene expression signature yielded P = 0.003. Real-time reverse transcription-PCR analysis of the four genes in an independent primary tumor set resulted in correct prediction of future relapse with an accuracy of 92%. One of the four genes in the prognostic signature, CCAAT/enhancer binding protein beta (C/EBPB), is expressed at higher levels in both primary relapsing tumors and metastatic tumors than in primary nonrelapsing tumors. Short interfering RNA-mediated down regulation of C/EBPB expression in WiT49, a cell line derived from a metastatic Wilms tumor, resulted in spontaneous apoptosis. These findings suggest that C/EBPB is a critical survival factor for Wilms tumor cells and that its expression contributes to the prognosis of Wilms tumor patients. PMID- 15805256 TI - Evidence against a role for SV40 in human mesothelioma. AB - SV40 has been implicated in the etiology of 40% to 60% of human mesotheliomas. These studies could have important medical implications concerning possible sources of human infection and potential therapies if human tumors are induced by this agent. We did PCR-based analysis to detect SV40 large T antigen DNA in human mesotheliomas. None of 69 tumors in which a single copy gene was readily amplified contained detectable SV40 large T antigen sequences. Under these conditions, it was possible to detect one copy of integrated SV40 DNA per cell in a mixture containing a 5,000-fold excess of normal cells using formalin-fixed preparations. Kidney, a known reservoir of SV40 in monkeys, from some of these individuals were also negative for SV40 large T antigen sequences. A subset of mesotheliomas was analyzed for SV40 large T antigen expression by immunostaining with a highly specific SV40 antibody. These tumors as well as several human mesothelioma cell lines previously reported to contain SV40 large T antigen were negative for detection of the virally encoded oncoprotein. Moreover, mesothelioma cell lines with wild-type p53 showed normal p53 function in response to genotoxic stress, findings inconsistent with p53 inactivation by the putative presence of SV40 large T antigen. Taken together, these findings strongly argue against a role of SV40 by any known transformation mechanism in the etiology of the majority of human malignant mesotheliomas. PMID- 15805257 TI - Linkage and microarray analyses of susceptibility genes in ACI/Seg rats: a model for prostate cancers in the aged. AB - ACI/Seg (ACI) rats develop prostate cancers spontaneously with aging, similar to humans. Here, to identify genes involved in prostate cancer susceptibility, we did linkage analysis and oligonucleotide microarray analysis. Linkage analysis was done using 118 effective rats, and prostate cancer susceptibility 1 (Pcs1), whose ACI allele dominantly induced prostate cancers, was mapped on chromosome 19 [logarithm of odds (LOD) score of 5.0]. PC resistance 1 (Pcr1), whose ACI allele dominantly and paradoxically suppressed the size of prostate cancers, was mapped on chromosome 2 (LOD score of 5.0). When linkage analysis was done in 51 rats with single or no macroscopic testicular tumors, which had larger prostates and higher testosterone levels than those with bilateral testicular tumors, Pcs2 and Pcr2 were mapped on chromosomes 20 and 1, respectively. By oligonucleotide microarray analysis with 8,800 probe sets and confirmation by quantitative reverse transcription-PCR, only two genes within these four loci were found to be differentially expressed >1.8-fold. Membrane metalloendopeptidase (Mme), known to inhibit androgen-independent growth of prostate cancers, on Pcr1 was expressed 2.0- to 5.5-fold higher in the ACI prostate, in accordance with its paradoxical effect. Cdkn1a on Pcs2 was expressed 1.5- to 4.5-fold lower in the ACI prostate. Additionally, genes responsible for testicular tumors and unilateral renal agenesis were mapped on chromosomes 11 and 14, respectively. These results showed that prostate cancer susceptibility of ACI rats involves at least four loci, and suggested Mme and Cdkn1a as candidates for Pcr1 and Pcs2. PMID- 15805259 TI - A lack of DNA mismatch repair on an athymic murine background predisposes to hematologic malignancy. AB - Inheritance of a germline mutation in one of the DNA mismatch repair genes predisposes human individuals to hereditary nonpolyposis colorectal cancer, characterized by development of tumors predominantly in the colon, endometrium, and gastrointestinal tract. Mice heterozygous for a mismatch repair-null mutation generally do not have an increased risk of neoplasia. However, mice constitutively lacking mismatch repair are prone to tumor development from an early age, particularly thymic lymphomas. Mismatch repair-deficient mice crossed to Apc(+/-) mice develop an increased spontaneous intestinal tumor incidence, demonstrating that the tumor spectrum can be genetically influenced. Here, we bred Msh2- and Msh6-deficient mice to athymic nude mice, hypothesizing that a broader tumor spectrum may be observed if mice are able to survive longer without succumbing to thymic lymphomas. However, Msh2(-/-);Foxn1(nu/nu) and Msh6(-/ );Foxn1(nu/nu) mice developed primarily early-onset lymphoblastic lymphomas. Using B-cell-specific markers, we found these tumors to be predominately B-cell in origin. The development of hematologic malignancy in the mouse, even in the absence of a thymus, parallels the development of B- and T-cell lymphoma and leukemia in the few rare mismatch repair-null human patients that have been identified. The persistent development of hematologic malignancy both in the mouse and in human patients deficient in mismatch repair leads us to implicate mismatch repair as an important repair mechanism in normal B- and T-cell development. Thus, mismatch repair-deficient mice may prove to be a good model to study human hematologic malignancy. PMID- 15805258 TI - Down-regulation of CD9 in human ovarian carcinoma cell might contribute to peritoneal dissemination: morphologic alteration and reduced expression of beta1 integrin subsets. AB - Peritoneal dissemination is one of the main causes of death in cancer patients. Pathophysiology of metastasis has been well investigated, but the mechanism of diffuse spread of tumor colonies in the peritoneal cavity is not fully understood. CD9 is a member of tetraspanin and its down-regulation is known to be involved in poor prognosis. To investigate the significance of the down regulation of CD9, HTOA, an ovarian carcinoma cell line that highly expressed CD9, was transiently transfected with small interfering RNA (siRNA) against CD9, and CD9-negative cells (HTOA(CD9-)) were purified. HTOA(CD9-) showed altered adhesion patterns on Matrigel, collagen, fibronectin, and laminin compared with those of control siRNA-transfected HTOA (control-HTOA). Flow cytometry and fluorescence cytostainings revealed that the expression levels of integrins beta1, alpha2, alpha3beta1, alpha5, and alpha6 were lower in HTOA(CD9-) than those of control-HTOA. HTOA(CD9-) showed altered expression of junctional and cytoskeletal molecules. By time-lapse video microscopy, control-HTOA showed solid adhesion to extracellular matrix and formed cobblestone pattern, whereas HTOA(CD9 ) showed weaker adhesion and were distributed as diffuse spots. To examine whether the expression level of CD9 change during tumor dissemination, HTOA-P, a highly disseminative subclone of HTOA, was established. HTOA-P showed distinctive down-regulation of CD9 at mRNA and protein levels, and showed similar morphologic alteration as HTOA(CD9-) did. These findings indicate that the down-regulation of CD9 may be an acquired event in the process of tumor dissemination. Down regulated CD9 may attenuate the expression of several integrins and rearrange junctional and cytoskeletal molecules that might contribute to dissemination of ovarian carcinomas. PMID- 15805260 TI - Cytoplasmic phospholipase A2 deletion enhances colon tumorigenesis. AB - Cellular pools of free arachidonic acid are tightly controlled through enzymatic release of the fatty acid and subsequent utilization by downstream enzymes including the cyclooxygenases. Arachidonic acid cleavage from membrane phospholipids is accomplished by the actions of phospholipase A(2) (PLA(2)). Upon release, free arachidonic acid provides substrate for the synthesis of eicosanoids. However, under certain conditions, arachidonic acid may participate in ceramide-mediated apoptosis. Disruption of arachidonic acid homeostasis can shift the balance of cell turnover in favor of tumorigenesis, via overproduction of tumor-promoting eicosanoids or alternatively by limiting proapoptotic signals. In the following study, we evaluated the influence of genetic deletion of a key intracellular phospholipase, cytoplasmic PLA(2) (cPLA(2)), on azoxymethane induced colon tumorigenesis. Heterozygous and null mice, upon treatment with the organotropic colon carcinogen, azoxymethane, developed a significant (P < 0.05) increase in colon tumor multiplicity (7.2-fold and 5.5-fold, respectively) relative to their wild-type littermates. This enhanced tumor sensitivity may be explained, in part, by the attenuated levels of apoptosis observed by terminal deoxynucleotidyl transferase-mediated nick end labeling staining within the colonic epithelium of heterozygous and null mice ( approximately 50% of wild type). The lower frequency of apoptotic cells corresponded with reduced ceramide levels (69% and 46% of wild-type littermates, respectively). Remarkably, increased tumorigenesis resulting from cPLA(2) deletion occurred despite a significant reduction in prostaglandin E(2) production, even in cyclooxygenase-2 overexpressing tumors. These data contribute new information that supports a fundamental role of cPLA(2) in the control of arachidonic acid homeostasis and cell turnover. Our findings indicate that the proapoptotic role of cPLA(2) in the colon may supercede its contribution to eicosanoid production in tumor development. PMID- 15805261 TI - Environmental pollutant and potent mutagen 3-nitrobenzanthrone forms DNA adducts after reduction by NAD(P)H:quinone oxidoreductase and conjugation by acetyltransferases and sulfotransferases in human hepatic cytosols. AB - 3-Nitrobenzanthrone (3-nitro-7H-benz[de]anthracen-7-one, 3-NBA) is a potent mutagen and suspected human carcinogen identified in diesel exhaust and air pollution. We compared the ability of human hepatic cytosolic samples to catalyze DNA adduct formation by 3-NBA. Using the (32)P-postlabeling method, we found that 12/12 hepatic cytosols activated 3-NBA to form multiple DNA adducts similar to those formed in vivo in rodents. By comparing 3-NBA-DNA adduct formation in the presence of cofactors of NAD(P)H:quinone oxidoreductase (NQO1) and xanthine oxidase, most of the reductive activation of 3-NBA in human hepatic cytosols was attributed to NQO1. Inhibition of adduct formation by dicoumarol, an NQO1 inhibitor, supported this finding and was confirmed with human recombinant NQO1. When cofactors of N,O-acetyltransferases (NAT) and sulfotransferases (SULT) were added to cytosolic samples, 3-NBA-DNA adduct formation increased 10- to 35-fold. Using human recombinant NQO1 and NATs or SULTs, we found that mainly NAT2, followed by SULT1A2, NAT1, and, to a lesser extent, SULT1A1 activate 3-NBA. We also evaluated the role of hepatic NADPH:cytochrome P450 oxidoreductase (POR) in the activation of 3-NBA in vivo by treating hepatic POR-null mice and wild-type littermates i.p. with 0.2 or 2 mg/kg body weight of 3-NBA. No difference in DNA binding was found in any tissue examined (liver, lung, kidney, bladder, and colon) between null and wild-type mice, indicating that 3-NBA is predominantly activated by cytosolic nitroreductases rather than microsomal POR. Collectively, these results show the role of human hepatic NQO1 to reduce 3-NBA to species being further activated by NATs and SULTs. PMID- 15805262 TI - Comprehensive DNA copy number profiling of meningioma using a chromosome 1 tiling path microarray identifies novel candidate tumor suppressor loci. AB - Meningiomas are common neoplasms of the meninges lining of the central nervous system. Deletions of 1p have been established as important for the initiation and/or progression of meningioma. The rationale of this array-CGH study was to characterize copy number imbalances of chromosome 1 in meningioma, using a full coverage genomic microarray containing 2,118 distinct measurement points. In total, 82 meningiomas were analyzed, making this the most detailed analysis of chromosome 1 in a comprehensive series of tumors. We detected a broad range of aberrations, such as deletions and/or gains of various sizes. Deletions were the predominant finding and ranged from monosomy to a 3.5-Mb terminal 1p homozygous deletion. Although multiple aberrations were observed across chromosome 1, every meningioma in which imbalances were detected harbored 1p deletions. Tumor heterogeneity was also observed in three recurrent meningiomas, which most likely reflects a progressive loss of chromosomal segments at different stages of tumor development. The distribution of aberrations supports the existence of at least four candidate loci on chromosome 1, which are important for meningioma tumorigenesis. In one of these regions, our results already allow the analysis of a number of candidate genes. In a large series of cases, we observed an association between the presence of segmental duplications and deletion breakpoints, which suggests their role in the generation of these tumor-specific aberrations. As 1p is the site of the genome most frequently affected by tumor specific aberrations, our results indicate loci of general importance for cancer development and progression. PMID- 15805263 TI - The t(8;9)(p22;p24) is a recurrent abnormality in chronic and acute leukemia that fuses PCM1 to JAK2. AB - We have identified a t(8;9)(p21-23;p23-24) in seven male patients (mean age 50, range 32-74) with diverse hematologic malignancies and clinical outcomes: atypical chronic myeloid leukemia/chronic eosinophilic leukemia (n = 5), secondary acute myeloid leukemia (n = 1), and pre-B-cell acute lymphoblastic leukemia (n = 1). Initial fluorescence in situ hybridization studies of one patient indicated that the nonreceptor tyrosine kinase Janus-activated kinase 2 (JAK2) at 9p24 was disrupted. Rapid amplification of cDNA ends-PCR identified the 8p22 partner gene as human autoantigen pericentriolar material (PCM1), a gene encoding a large centrosomal protein with multiple coiled-coil domains. Reverse transcription-PCR and fluorescence in situ hybridization confirmed the fusion in this case and also identified PCM1-JAK2 in the six other t(8;9) patients. The breakpoints were variable in both genes, but in all cases the chimeric mRNA is predicted to encode a protein that retains several of the predicted coiled-coil domains from PCM1 and the entire tyrosine kinase domain of JAK2. Reciprocal JAK2 PCM1 mRNA was not detected in any patient. We conclude that human autoantigen pericentriolar material (PCM1)-JAK2 is a novel, recurrent fusion gene in hematologic malignancies. Patients with PCM1-JAK2 disease are attractive candidates for targeted signal transduction therapy. PMID- 15805264 TI - Gene amplification is a mechanism of Six1 overexpression in breast cancer. AB - The Six1 homeoprotein plays a critical role in expanding progenitor populations during normal development via its stimulation of proliferation and inhibition of apoptosis. Overexpression of Six1 is observed in several tumor types, suggesting that when expressed out of context, Six1 may contribute to tumorigenesis by reinstating properties normally conveyed on developing cells. Indeed, Six1 contributes to tumor cell proliferation both in breast cancer and in rhabdomyosarcomas, in which it is also implicated in metastasis. Whereas Six1 overexpression has been reported in several tumor types, the mechanism responsible for its overexpression has not previously been examined. Here we show that a change in gene dosage may contribute to Six1 mRNA overexpression. Significant Six1 gene amplification and overrepresentation occurs in numerous breast cancer cell lines as compared with normal mammary epithelial cells, and the changes in gene dosage correlate with increased Six1 mRNA levels. Of 214 human infiltrating ductal breast carcinomas examined for Six1 gene dosage, 4.7% show Six1 amplification/overrepresentation, and tumors that exhibit an increase in Six1 gene dosage overexpress Six1 mRNA. These data implicate Six1 gene amplification/overrepresentation as a mechanism of Six1 mRNA overexpression in human breast cancer. PMID- 15805265 TI - p210BCR-ABL inhibits SDF-1 chemotactic response via alteration of CXCR4 signaling and down-regulation of CXCR4 expression. AB - It has been shown that p210(BCR-ABL) significantly impairs CXCR4 signaling. We report here that the migratory response to SDF-1 was profoundly altered in blast crisis, whereas chronic-phase CD34(+) cells migrated normally to this chemokine. This migratory defect was associated with a low CXCR4 membrane expression. In vitro STI-571 treatment of CD34(+) cells from patients in blast crisis markedly increased the CXCR4 transcript and CXCR4 membrane expression. Because p210(BCR ABL) frequently increases with disease progression, we determined the effects of high and low p210(BCR-ABL) expression on CXCR4 protein in the granulocyte macrophage colony-stimulating factor-dependent human cell line MO7e. p210(BCR ABL) expression distinctly alters CXCR4 protein through two different mechanisms depending on its expression level. At low expression, a signaling defect was detected with no modification of CXCR4 expression. However, higher p210(BCR-ABL) expression induced a marked down-regulation of CXCR4 that is related to its decreased transcription. The effect of p210(BCR-ABL) required its tyrosine kinase activity. Collectively, these data indicate that p210(BCR-ABL) could affect CXCR4 by more than one mechanism and suggest that down-regulation of CXCR4 may have important implications in chronic myelogenous leukemia pathogenesis. PMID- 15805266 TI - Histone deacetylase inhibitors decrease DNA methyltransferase-3B messenger RNA stability and down-regulate de novo DNA methyltransferase activity in human endometrial cells. AB - It is well known that the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) acts synergistically with the DNA methyltransferase (DNMT) inhibitor 5-aza 2'-deoxycytidine (ADC) to reactivate DNA methylation-silenced genes. Moreover, in several studies, TSA was capable of inducing DNA demethylation even in the absence of ADC. Here we describe a mechanism by which HDAC inhibitors affect DNA methylation through their regulation on DNMT3B, a methyltransferase responsible for de novo DNA methylation. Using quantitative real-time PCR and Western blot analysis, we show that TSA down-regulates DNMT3B mRNA and protein expression in human endometrial cancer cells. This decrease in DNMT3B mRNA results in a significant reduction in de novo methylation activities. Further experiments indicated that TSA decreases DNMT3B mRNA stability and reduces its half-life from approximately 4 to 2.5 hours. We established that protein synthesis is required for posttranscriptional regulation, suggesting the involvement of an RNase and/or key mRNA stabilization factor(s) controlling the DNMT3B mRNA stability. Therefore, TSA may not only modify histone acetylation, but also potentially alter DNA methylation. Since the HDAC inhibitors are frequently used in epigenetic studies and are considered to be promising anticancer drugs, these new findings will have implications in both laboratory and clinical settings. PMID- 15805267 TI - Transcriptional regulation of cyclin A2 by RASSF1A through the enhanced binding of p120E4F to the cyclin A2 promoter. AB - Recent advances in the study of RASSF1A, the candidate tumor suppressor gene, indicate a possible role of RASSF1A in cell cycle regulation; however, very little is known regarding molecular mechanisms underlying this control. Using small interfering RNA to knockdown endogenous RASSF1A in the breast tumor cell line HB2 and in the cervical cancer cell line HeLa, we identify that a key player in cell cycle progression, cyclin A2, is concomitantly increased at both protein and mRNA levels. In A549 clones stably expressing RASSF1A, cyclin A2 levels were diminished compared with vector control. A known transcriptional regulator of cyclin A2, p120(E4F) (a repressor of cyclin A2), has been shown previously by our group to interact with RASSF1A. We show that levels of p120(E4F) are not affected by RASSF1A small interfering RNA in HB2 and HeLa cells. However, electrophoretic mobility shift assays indicate that knockdown of endogenous RASSF1A in HB2 and HeLa cells leads to a reduction in the binding capacity of p120(E4F) to the cyclin A2 promoter, whereas in the A549 clone stably expressing RASSF1A the binding capacity is increased. These data are further corroborated in vitro by the luciferase assay and in vivo by chromatin immunoprecipitation experiments. Together, these data identify the cyclin A2 gene as a cellular target for RASSF1A through p120(E4F) and for the first time suggest a transcriptional mechanism for RASSF1A-dependent cell cycle regulation. PMID- 15805268 TI - Small interfering RNA-mediated Polo-like kinase 1 depletion preferentially reduces the survival of p53-defective, oncogenic transformed cells and inhibits tumor growth in animals. AB - Polo-like kinase 1 (Plk1) is required for multiple stages of mitosis and is up regulated in many human malignancies. We depleted Plk1 expression using small interfering RNA (siRNA) and showed defects in bipolar spindle formation and cytokinesis, growth inhibition, and apoptosis induction in human cancer cell lines. To our surprise, depletion of Plk1 in normal human cells did not result in obvious cell cycle defects, and did not induce significant inhibition of cell growth for at least two cell cycles. In addition, Plk1 siRNA inhibited colony formation in soft agar and tumorigenesis in a HT1080 xenograft model in a dose dependent manner. Analysis with isogenic pairs of cell lines, differing in p53 status, revealed that Plk1 depletion preferentially induced mitotic arrest, aneuploidy, and reduced cell survival in the p53-defective cell lines. No obvious defects were observed in most p53 wild-type cells during the first few cell cycles. In addition, long-term survival studies revealed that p53 facilitates survival upon Plk1 depletion. Therefore, short-term inhibition of Plk1 can kill tumor cells while allowing normal cells to survive. These data validate the episodic inhibition of Plk1 as a very useful approach for cancer treatment. PMID- 15805269 TI - Down-regulation of Cx43 by retroviral delivery of small interfering RNA promotes an aggressive breast cancer cell phenotype. AB - Connexins are gap junction proteins that assemble into channels that mediate direct intercellular communication. Connexins are well-documented tumor suppressors and are thought to regulate both cell growth and differentiation. As previously reported, most human breast tumors and cell lines down-regulate gap junctions or have defective gap junctional intercellular communication. Furthermore, overexpression of connexins in breast cancer cells inhibits tumor growth in vivo. In this study, we hypothesize that controlled Cx43 down regulation would induce breast tumor cells to acquire a more aggressive phenotype. Here we report that Cx43 was down-regulated in both normal rat kidney (NRK) cells and human breast cancer cell lines (MDA-MB-231 and Hs578T) by transfection with chemically synthesized small interfering RNA (siRNA) or short hairpin RNA generated from a retroviral infection. Furthermore, we show that retroviral delivery and expression of siRNA directed to different coding regions of Cx43 resulted in differential levels of Cx43 silencing and impaired gap junctional intercellular communication. Cx43-silenced Hs578T cells grew faster and were more migratory. Finally, Western blot analysis revealed that down regulation of Cx43 resulted in decreased expression of thrombospondin-1, an antiangiogenesis molecule, and increased expression of vascular endothelial growth factor. Taken together, these results suggest that Cx43 is required for maintaining cell differentiation and the regulation of molecules important in angiogenesis. PMID- 15805270 TI - Rapid access of antibodies to alpha5beta1 integrin overexpressed on the luminal surface of tumor blood vessels. AB - Integrin alpha(5)beta(1) is overexpressed on endothelial cells of tumor vessels and is uniformly and rapidly accessible to antibodies in the bloodstream. Here, we determined whether antibodies rapidly gain access to integrin overexpressed on the abluminal (basolateral) surface of endothelial cells through vascular leakiness or whether the rapid accessibility results instead because the integrin is overexpressed on the luminal (apical) surface of endothelial cells due to loss of cell polarity. Using tumors in RIP-Tag2 transgenic mice as a model, we first compared the binding pattern of intravascular anti-alpha(5)beta(1) integrin antibody with the leakage pattern of nonspecific IgG. The distributions did not match: anti-alpha(5)beta(1) integrin antibody uniformly labeled the tumor vasculature, but IgG was located in patchy sites of leakage. We next injected an antibody to fibrinogen/fibrin, which resulted in patchy labeling of tumors that matched the leakage of IgG and the overall distribution of fibrin in tumors. Similarly, injected antibodies to the basement membrane protein fibronectin, a ligand of alpha(5)beta(1) integrin, or type IV collagen produced patchy sites of leakage instead of uniform labeling of vascular basement membrane. Differences in the kinetics of labeling, which for alpha(5)beta(1) integrin antibody was near maximal by 10 minutes but for the other antibodies gradually increased over 6 hours, indicated differences in accessibility of their respective targets. Isosurface rendering of confocal microscopic images was consistent with antibody binding to alpha(5)beta(1) integrin on the luminal surface of endothelial cells. Together, these findings indicate that the rapid accessibility of alpha(5)beta(1) integrin in RIP-Tag2 tumors results from overexpression of the integrin on the luminal surface of tumor vessels. PMID- 15805271 TI - Telomerase-independent telomere length maintenance in the absence of alternative lengthening of telomeres-associated promyelocytic leukemia bodies. AB - Immortal tumor cells and cell lines employ a telomere maintenance mechanism that allows them to escape the normal limits on proliferative potential. In the absence of telomerase, telomere length may be maintained by an alternative lengthening of telomeres (ALT) mechanism. All human ALT cell lines described thus far have nuclear domains of unknown function, termed ALT-associated promyelocytic leukemia bodies (APB), containing promyelocytic leukemia protein, telomeric DNA and telomere binding proteins. Here we describe telomerase-negative human cells with telomeres that contain a substantial proportion of nontelomeric DNA sequences (like telomerase-null Saccharomyces cerevisiae survivor type I cells) and that are maintained in the absence of APBs. In other respects, they resemble typical ALT cell lines: the telomeres are highly heterogeneous in length (ranging from very short to very long) and undergo rapid changes in length. In addition, these cells are capable of copying a targeted DNA tag from one telomere into other telomeres. These data show that APBs are not always essential for ALT mediated telomere maintenance. PMID- 15805272 TI - A novel telomere structure in a human alternative lengthening of telomeres cell line. AB - Cancer cells require mechanisms to maintain telomeres. Most use telomerase, but 5% to 20% of tumors use alternative lengthening of telomeres (ALT), a telomerase independent mechanism that seems to depend on recombination. ALT is characterized by amplification of telomere TTAGGG repeats to lengths beyond 50 kb, by elevated rates of telomere recombination, and by nuclear structures called ALT-associated promyelocytic leukemia bodies. In Saccharomyces cerevisiae, survivors of telomerase inactivation also use recombination to maintain telomeres. There are two types of survivors, which differ in telomere structure. The first possesses telomere repeats and the Y' subtelomeric element amplified together as a tandem array at chromosome termini (type I), and the other possesses amplification of telomeric repeats alone (type II), similar to previously described human ALT cells. Here, we describe the first human ALT cell line having "tandem array" telomeres with a structure similar to that of type I yeast survivors. The chromosome termini consist of a repeat unit containing approximately 2.5 kb of SV40 DNA and a variable amount of TTAGGG sequence repeated in tandem an average of 10 to 20 times. Similar to previously described ALT cells, they show evidence of telomere recombination, but unlike standard ALT cells, they lack ALT associated promyelocytic leukemia bodies and their telomeres are transcribed. These findings have implications for the pathogenesis and diagnosis of cancer. PMID- 15805273 TI - Broad-spectrum G protein-coupled receptor antagonist, [D-Arg1,D Trp5,7,9,Leu11]SP: a dual inhibitor of growth and angiogenesis in pancreatic cancer. AB - Substance P analogues, including [D-Arg(1),D-Trp(5,7,9),Leu(11)]SP (SPA) are broad-spectrum G protein-coupled receptor (GPCR) antagonists that have potential antitumorigenic activities, although the mechanism(s) are not completely understood. Here, we examined the effects of SPA in ductal pancreatic cancers that express multiple GPCRs for mitogenic agonists and also produce proangiogenic chemokines. Using HPAF-II, a well-differentiated pancreatic cancer cell line as our model system, we showed that SPA inhibited multiple neuropeptide-induced Ca(2+) mobilization, DNA synthesis, and anchorage-independent growth in vitro. SPA also significantly attenuated the growth of HPAF-II tumor xenografts in nude mice beyond the treatment period. Interestingly, SPA markedly increased apoptosis but moderately decreased proliferation marker, Ki-67 in the tumor xenografts implying additional mechanism(s) for the significant growth inhibitory effect observed in vivo. HPAF-II cells express ELR(+) CXC chemokines, including IL 8/CXCL8, which bind to CXCR2 (a member of GPCR superfamily) and promote angiogenesis in multiple cancers, including pancreatic cancer. SPA inhibited CXCR2-mediated Ca(2+) mobilization and blocked specifically IL-8/CXCL8-induced angiogenesis in rat corneal micropocket assay in vivo. A salient feature of the results presented here is that SPA markedly reduced tumor-associated angiogenesis in the HPAF-II xenografts in vivo. Our results show that SPA, a broad-spectrum GPCR antagonist attenuates tumor growth in pancreatic cancer via a dual mechanism involving both the antiproliferative and antiangiogenic properties. We conclude that this novel dual-inhibitory property of SPA could be of significant therapeutic value in pancreatic cancer, when used in combination with other antiproliferative and/or antiangiogenic agents. PMID- 15805274 TI - Drastic down-regulation of Kruppel-like factor 4 expression is critical in human gastric cancer development and progression. AB - Kruppel-like factor 4 (KLF4) is highly expressed in epithelial tissues such as the gut and skin. However, the role of KLF4 in human gastric cancer development and progression is unknown. Here we show that KLF4 protein expression was decreased or lost in primary tumors and, in particular, lymph node metastases when compared with that in normal gastric mucosa. Moreover, loss of KLF4 expression in the primary tumors was significantly associated with poor survival, and also an independent prognostic marker in a multivariate analysis. Consistently, most human gastric cancer cell lines exhibited loss of or a substantial decrease in KLF4 expression at both RNA and protein levels. Enforced restoration of KLF4 expression resulted in marked cell growth inhibition in vitro and significantly attenuated tumor growth and total abrogation of metastasis in an orthotopic animal model of gastric cancer. Mechanism studies indicated that promoter hypermethylation and hemizygous deletion contributed to the down regulation of KLF4 expression and the induction of apoptosis contributed to the antitumor activity of KLF4. Collectively, our data provide first clinical and casual evidence and potential mechanism that the alteration of KLF4 expression plays a critical role in gastric cancer development and progression. PMID- 15805275 TI - Proteomic analysis reveals hyperactivation of the mammalian target of rapamycin pathway in neurofibromatosis 1-associated human and mouse brain tumors. AB - Individuals with the tumor predisposition syndrome, neurofibromatosis 1 (NF1), are prone to development of nervous system tumors, including neurofibromas and pilocytic astrocytomas. Based on the ability of the NF1 gene product (neurofibromin) to function as a GTPase activating protein for RAS, initial biologically based therapies for NF1-associated tumors focused on the use of RAS inhibitors, but with limited clinical success. In an effort to identify additional targets for therapeutic drug design in NF1, we used an unbiased proteomic approach to uncover unanticipated intracellular signaling pathways dysregulated in Nf1-deficient astrocytes. We found that the expression of proteins involved in promoting ribosome biogenesis was increased in the absence of neurofibromin. In addition, Nf1-deficient astrocytes exhibit high levels of mammalian target of rapamycin (mTOR) pathway activation, which was inhibited by blocking K-RAS or phosphatidylinositol 3-kinase activation. This mTOR pathway hyperactivation was reflected by high levels of ribosomal S6 activation in both Nf1 mutant mouse optic nerve gliomas and in human NF1-associated pilocytic astrocytoma tumors. Moreover, inhibition of mTOR signaling in Nf1-/- astrocytes abrogated their growth advantage in culture, restoring normal proliferative rates. These results suggest that mTOR pathway inhibition may represent a logical and tractable biologically based therapy for brain tumors in NF1. PMID- 15805276 TI - Hypoxia stimulates carcinoma invasion by stabilizing microtubules and promoting the Rab11 trafficking of the alpha6beta4 integrin. AB - Hypoxia plays a key role in tumor cell survival, invasion, and metastasis. Here we show that hypoxia increases tumor cell invasion by the modulation of Rab11, an important molecule for vesicular trafficking, especially membrane protein recycling and translocation of proteins from trans-Golgi network to plasma membrane. Dominant-negative Rab11 dramatically decreased hypoxia-induced invasion of MDA-MB-231 breast carcinoma cells without affecting cell apoptosis. Hypoxia induced Rab11 trafficking is regulated by microtubule stability, as evidenced by the findings that hypoxia increases Glu tubulin and that colchicine blocks Rab11 trafficking and invasion. Inhibition of GSK-3beta activity by hypoxia seems to be central to microtubule stabilization and invasion. In fact, expression of a dominant-negative GSK-3beta was sufficient to stimulate invasion in normoxia. One target of Rab11-mediated trafficking that contributes to invasion is the integrin alpha6beta4. Hypoxia induced a significant increase in alpha6beta4 surface expression but it had no effect on the surface expression of alpha3beta1. This increase is dependent on Rab11 and stable microtubules. In summary, we identify vesicle trafficking as a novel target of hypoxic stimulation that is important for tumor invasion. PMID- 15805277 TI - Signaling pathways associated with colonic mucosa hyperproliferation in mice overexpressing gastrin precursors. AB - MTI/G-Gly mice and hGAS mice, overexpressing glycine-extended gastrin (G-Gly) and progastrin, respectively, display colonic mucosa hyperplasia, hyperproliferation, and an increased susceptibility to intestinal neoplasia. Here, we have used these transgenic mice to analyze in vivo the modulation of intracellular signaling pathways that may be responsible for the proliferative effects of gastrin precursors. The expression, activation, and localization of signaling and cell-to cell adhesion molecules were studied using immunofluorescence and Western blot techniques on colonic tissues derived from MTI/G-Gly, hGAS, or wild-type FVB/N mice. These analyses revealed an up-regulation of Src tyrosine kinase and related signaling pathways [phosphatidyl inositol 3'-kinase (PI3K)/Akt, Janus-activated kinase (JAK) 2, signal transducer and activator of transcription (STAT) 3, and extracellular-signal regulated kinases (ERK)] in both MTI/G-Gly and hGAS mice compared with the wild-type control animals as well as an overexpression of transforming growth factor-alpha (TGF-alpha). In contrast, overexpression of the gastrin precursors did not affect the activation status of STAT1 nor the expression and the distribution of adhesion proteins (focal adhesion kinase, cadherins, and catenins). We report for the first time that the transition from a normal colonic epithelium to a hyperproliferative epithelium in MTI/G-Gly and hGAS mice may be a consequence of the up-regulation of Src, PI3K/Akt, JAK2, STAT3, ERKs, and TGF-alpha. Deregulation of cell adhesion, a late event in tumor progression, does not occur in these transgenic models. PMID- 15805278 TI - Reversible conversion of immortal human cells from telomerase-positive to telomerase-negative cells. AB - Immortal cell lines and tumors maintain their telomeres via the telomerase pathway or via a telomerase-independent pathway, referred to as alternative lengthening of telomeres (ALT). Here, we show the reversible conversion of the human papillomavirus type 16 E6-induced immortal human fibroblasts E6 Cl 6 from telomerase-positive (Tel(+)) to telomerase-negative (Tel(-)) cells. Tel(+) cells converted spontaneously to Tel(-) cells that reverted to Tel(+) cells following treatment with trichostatin A (TSA) and/or 5-aza-2'-deoxycytidine (5-AZC), which induced the reversion from complete to partial methylation of the CpG islands of the human telomerase reverse transcriptase (hTERT) promoter in Tel(-) E6 Cl 6 cells. Tel(-) E6 Cl 6 cells lacked the phenotypes characteristic of ALT cell lines such as very long and heterogenous telomeres and ALT-associated promyelocytic leukemia nuclear bodies (APB) but grew for >240 population doublings (PD) after they became telomerase negative. The ratios of histone H3 (H3) lysine (K) 9 methylation to each of H3-K4 methylation, H3-K9 acetylation, and H3-K14 acetylation of the chromatin containing the hTERT promoter in Tel(-) E6 Cl 6 cells and ALT cell lines were greater than those in Tel(+) cells and decreased following treatment with TSA and/or 5-AZC, inversely corresponding to telomerase activity. Our findings suggest the possibility that human tumors may be able to reversibly interconvert their telomere maintenance phenotypes by chromatin structure-mediated regulation of hTERT expression. PMID- 15805279 TI - p73-dependent apoptosis through death receptor: impairment by human cytomegalovirus infection. AB - The discovery of p73, a p53-related protein with various isotypes resulting from different promoter usage or splicing events, provided new insights into regulation of neurogenesis and tumorigenesis. Among p73 isoforms described thus far, TA-truncated molecules (DeltaN) appeared as key proteins according to their antagonistic activity against transcription factor activity of p53 family members. We previously showed that infection by human cytomegalovirus (HCMV) induced drug resistance and altered p53- and p73-dependent apoptosis of infected cells through accumulation of DeltaN-p73alpha. In accordance with the ability of p53 to induce apoptosis through death receptors, we asked whether p73 activation could compensate for p53 deficiency. We showed that p73 transcriptional activity sensitized cells to apoptosis through death receptors in a caspase-dependent pathway. Expression of the death-inducing signaling complex (DISC) proteins was unchanged, whereas p73 activation through either cisplatin treatment or ectopic overexpression induced up-regulation of Fas transcription and expression at cell surface. According to its ability to flood cells with DeltaN-p73alpha, HCMV inhibited p73-dependent Fas-mediated apoptosis, gaining an additional trick to favor its survival in the host cell. Owing to the involvement of p53- and p73 dependent death receptor signaling in development of the central nervous system, immune surveillance of neural cells, and sensitivity of tumors to drugs, our previous and present data prompt us to consider stabilization of DeltaN-p73alpha by HCMV as a possible mechanism in impairment of embryogenesis and in tumorigenesis. PMID- 15805280 TI - Escape from therapy-induced accelerated cellular senescence in p53-null lung cancer cells and in human lung cancers. AB - Accelerated cellular senescence (ACS) has been described for tumor cells treated with chemotherapy and radiation. Following exposure to genotoxins, tumor cells undergo terminal growth arrest and adopt morphologic and marker features suggestive of cellular senescence. ACS is elicited by a variety of chemotherapeutic agents in the p53-null, p16-deficient human non-small cell H1299 carcinoma cells. After 10 to 21 days, infrequent ACS cells (1 in 10(6)) can bypass replicative arrest and reenter cell cycle. These cells express senescence markers and resemble the parental cells in their transcription profile. We show that these escaped H1299 cells overexpress the cyclin-dependent kinase Cdc2/Cdk1. The escape from ACS can be disrupted by Cdc2/Cdk1 kinase inhibitors or by knockdown of Cdc2/Cdk1 with small interfering RNA and can be promoted by expression of exogenous Cdc2/Cdk1. We also present evidence that ACS occurs in vivo in human lung cancer following induction chemotherapy. Viable tumors following chemotherapy also overexpress Cdc2/Cdk1. We propose that ACS is a mechanism of in vivo tumor response and that mechanisms aberrantly up-regulate Cdc2/Cdk1 promotes escape from the senescence pathway may be involved in a subset of tumors and likely accounts for tumor recurrence/progression. PMID- 15805281 TI - Identification of the genes involved in enhanced fenretinide-induced apoptosis by parthenolide in human hepatoma cells. AB - Fenretinide (N-4-hydroxyphenyl retinamide, 4HPR) is a synthetic anticancer retinoid that is a well-known apoptosis-inducing agent. Recently, we observed that the apoptosis induced by fenretinide could be effectively enhanced in hepatoma cells by a concomitant treatment with parthenolide, which is a known inhibitor of nuclear factor-kappaB (NF-kappaB). Furthermore, treatment with fenretinide triggered the activation of NF-kappaB during apoptosis, which could be substantially inhibited by parthenolide, suggesting that NF-kappaB activation during fenretinide-induced apoptosis has an antiapoptotic effect. This study investigated the molecular mechanism of this apoptotic potentiation by NF-kappaB inhibition. The genes involved in the enhanced fenretinide-induced apoptosis by parthenolide were identified using the differential display-PCR method and subsequent Northern blot or semiquantitative reverse transcriptase PCR analysis. This study identified 35 apoptosis-related genes including 12 unknown genes that were either up- or down-regulated by parthenolide. Interestingly, one up regulated gene (HA1A2) was isolated and cloned from the liver cDNA, and was found to be identical to ANKRD1, which is also referred to as the CARP gene. Compared with controls treated with an empty vector or with antisense cDNA, the ectopic expression of ANKRD1 led to reduced colony formation and to enhanced apoptotic cell death in hepatoma cells. These results suggest that ANKRD1 and the other genes, whose expressions were substantially modulated by the parthenolide mediated inhibition of NF-kappaB activation, play roles in the enhanced drug induced apoptosis. In addition, this study suggests that those identified genes may be useful in anticancer strategies against hepatoma. PMID- 15805282 TI - Proapoptotic activity of cell-permeable anti-Akt single-chain antibodies. AB - We developed anti-Akt1 single-chain antibodies (scFv) by panning a mouse phage displayed scFv recombinant antibody library. Recombinant scFv that bound glutathione S-transferase (GST)-Akt1 were screened for their ability to inhibit Akt activity in vitro in a kinase reaction containing human recombinant Akt1 and an Akt/serum glucocorticoid-inducible kinase (SGK) substrate. Michaelis-Menten analysis of kinase inhibition by a selected scFv was consistent with scFv mediated competition with enzyme's substrate for the catalytic site of Akt. To generate a membrane-permeable version of the anti-Akt1 scFv, the scFv gene was subcloned into a GST expression vector carrying a membrane-translocating sequence (MTS) from Kaposi fibroblast growth factor. A purified GST-anti-Akt1-MTS fusion protein accumulated intracellularly in 293T, BT-474, and PyVmT cells in a dose- and time-dependent fashion. Intracellular accumulation correlated temporally with inhibition of p-Ser(473) Akt and GSK-3alpha/beta phosphorylation, suggesting that Ser(473) is an Akt autophosphorylation site. Phosphorylated (activated) phosphoinositide-dependent kinase 1, mitogen-activated protein kinase, p38, and HER2 (erbB2) were not affected, supporting Akt kinase specificity for the inhibitory scFv. Exogenously expressed constitutively active Akt2 and Akt3 were also inhibited in vitro by the anti-Akt1 fusion protein. Furthermore, GST-anti Akt1-MTS induced apoptosis in three cancer cell lines that express constitutively active Akt. Finally, systemic treatment with the anti-Akt scFv reduced tumor volume and neovascularization and increased apoptosis in PyVmT-expressing transgenic tumors implanted in mouse dorsal window chambers. Thus, GST-anti-Akt1 MTS is a novel cell-permeable inhibitor of Akt, which selectively inhibits Akt mediated survival in intact cells both in vitro and in vivo. PMID- 15805283 TI - Effects of the mammalian target of rapamycin inhibitor CCI-779 used alone or with chemotherapy on human prostate cancer cells and xenografts. AB - Selective inhibition of repopulation of surviving tumor cells between courses of chemotherapy might improve the outcome of treatment. A potential target for inhibiting repopulation is the mammalian target of rapamycin pathway; PTEN negative tumor cells are particularly sensitive to inhibition of this pathway. Here we study the rapamycin analogue CCI-779, alone or with chemotherapy, as an inhibitor of proliferation of the human prostate cancer cell lines PC-3 and DU145. The PTEN and phospho-Akt/PKB status and the effect of CCI-779 on phosphorylation of ribosomal protein S6 were evaluated by immunostaining and/or Western blotting. Expression of phospho-Akt/PKB in PTEN mutant PC-3 cells and xenografts was higher than in PTEN wild-type DU145 cells. Phosphorylation of S6 was inhibited by CCI-779 in both cell lines. Cultured cells were treated weekly with mitoxantrone or docetaxel for two cycles, and CCI-779 or vehicle was given between courses. Growth and clonogenic survival of both cell lines were inhibited in a dose-dependent manner by CCI-779, but there were minimal effects when CCI 779 was given between courses of chemotherapy. CCI-779 inhibited the growth of xenografts derived from both cell lines with greater effects against PC-3 than DU145 tumors. CCI-779 caused mild myelosuppression. The activity of mitoxantrone or docetaxel was limited, but CCI-779 given between courses of chemotherapy increased growth delay of PC-3 xenografts. Our results suggest that repopulation of PTEN-negative cancer cells between courses of chemotherapy might be inhibited by CCI-779. PMID- 15805284 TI - An oncolytic HSV-1 mutant expressing ICP34.5 under control of a nestin promoter increases survival of animals even when symptomatic from a brain tumor. AB - Oncolytic herpes simplex virus-1 (HSV-1) mutants possessing mutations in the ICP34.5 and ICP6 genes have proven safe through clinical trials. However, ICP34.5 null viruses may grow poorly in cells due to their inability to prevent host-cell shut-off of protein synthesis caused by hyperphosphorylation of eukaryotic initiation factor 2alpha. To increase tumor selectivity, glioma-selective expression of ICP34.5 in the context of oncolysis may be useful. Malignant gliomas remain an incurable disease. One molecular marker of malignant gliomas is expression of the intermediate filament nestin. Expression of nestin mRNA was confirmed in 6 of 6 human glioma lines and in 3 of 4 primary glioma cells. Normal human astrocytes were negative. A novel glioma-selective HSV-1 mutant (rQNestin34.5) was thus engineered by expressing ICP34.5 under control of a synthetic nestin promoter. Replication, cellular propagation, and cytotoxicity of rQNestin34.5 were significantly enhanced in cultured and primary human glioma cell lines compared with control virus. However, replication, cellular propagation, and cytotoxicity of rQNestin34.5 in normal human astrocytes remained quantitatively similar to that of control virus. In glioma cell lines infected with rQNestin34.5, the level of phospho-eukaryotic initiation factor 2alpha was lower than that of cells infected by control rHsvQ1, confirming selective ICP34.5 expression in glioma cells. In vivo, rQNestin34.5 showed significantly more potent inhibition of tumor growth compared with control virus. Treatment in the brain tumor model was instituted on animal's display of neurologic symptoms, which usually led to rapid demise. rQNestin34.5 treatment doubled the life span of these animals. These results show that rQNestin34.5 could be a potent agent for the treatment of malignant glioma. PMID- 15805285 TI - Carboxyl-terminal Src kinase homologous kinase negatively regulates the chemokine receptor CXCR4 through YY1 and impairs CXCR4/CXCL12 (SDF-1alpha)-mediated breast cancer cell migration. AB - Using microarray gene analysis, we found that carboxyl-terminal Src kinase homologous kinase (CHK) regulated the expression of the chemokine receptor, CXCR4. Northern blot and fluorescence-activated cell-sorting analyses showed that CHK down-regulated CXCR4 mRNA and protein levels, respectively. Mutated CHK, which contains a mutation within the ATP binding site of CHK, failed to inhibit CXCR4 expression, thus suggesting that CHK kinase activity is involved in the regulation of CXCR4. Results from gel shift analysis indicated that CHK regulates CXCR4 transcriptional activity by altering YY1 binding to the CXCR4 promoter. Whereas CHK had no significant effects on the expression of YY1, c-Myc, Max, and other YY1-binding proteins, CHK was found to modulate the YY1/c-Myc association. Furthermore, CHK inhibited CXCR4-positive breast cancer cell migration. Taken together, these studies show a novel mechanism by which CHK down-regulates CXCR4 through the YY1 transcription factor, leading to decreased CXCR4-mediated breast cancer cell motility and migration. PMID- 15805286 TI - Targeting tomoregulin for radioimmunotherapy of prostate cancer. AB - Radiotherapy is an effective approach for the treatment of local prostate cancer. However, once prostate cancer metastasizes, radiotherapy cannot be used due to the distribution of multiple metastases to lymph nodes and bones. In contrast, radioimmunotherapy should still be efficacious in metastatic prostate cancer as radioisotopes are brought to tumor cells by targeting antibodies. Here we identify and validate a prostate-expressed molecule, tomoregulin, as a target for radioimmunotherapy of prostate cancer. Tomoregulin is a transmembrane protein selectively expressed in the brain, prostate, and prostate cancer, but not expressed in other normal tissues. Immunohistochemical studies of tomoregulin protein in clinical samples show its location in the luminal epithelium of normal prostate, benign prostatic hyperplasia, and prostatic intraepithelial neoplasia. More importantly, the tomoregulin protein is expressed in primary prostate tumors and in their lymph node and bone metastases. The nature of tomoregulin as a transmembrane protein and its tissue-specific expression make tomoregulin an attractive target for radioimmunotherapy, in which tomoregulin-specific antibodies will deliver a radioisotope to prostate tumor cells and metastases. Indeed, biodistribution studies using a prostate tumor xenograft model showed that the (111)In-labeled anti-tomoregulin antibody 2H8 specifically recognizes tomoregulin protein in vivo, leading to a strong tumor-specific accumulation of the antibody. In efficacy studies, a single i.p. dose of 150 microCi (163 microg) (90)Y-labeled 2H8 substantially inhibits the growth rate of established LNCaP human prostate tumor xenograft in nude mice but produces no overt toxicity despite cross-reactivity of 2H8 with mouse tomoregulin. Our data clearly validate tomoregulin as a target for radioimmunotherapy of prostate cancer. PMID- 15805287 TI - Enhancement of the therapeutic efficacy of taxol by the mitogen-activated protein kinase kinase inhibitor CI-1040 in nude mice bearing human heterotransplants. AB - Taxol may contribute to intrinsic chemoresistance by activating the mitogen activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) cytoprotective pathway in human cancer cell lines and tumors. We have previously shown additivity between Taxol and the MEK inhibitor, U0126 in human cancer cell lines. Here, the combination of Taxol with an orally bioavailable MEK inhibitor, CI-1040, was evaluated in human lung tumors heterotransplanted into nude mice. Unlike xenograft models that are derived from cells with multiple genetic alterations due to prolonged passage, heterotransplanted tumor models are more clinically relevant. Combined treatment with both drugs resulted in inhibition of tumor growth in all models and tumor regressions in three of four models tested, supporting our previous observation that Taxol's efficacy is potentiated by MEK inhibition. Concurrent administration was superior to intermittent dosing. Pharmacodynamic assessments of tumors indicated that suppression of MEK was associated with induction of S473 phosphorylated Akt and reduced proliferation in the combination groups relative to single agents, in addition to suppression of fibroblast growth factor-mediated angiogenesis and reduced expression of vascular endothelial growth factor. These findings are significant and indicate that this combination may have broad therapeutic applications in a diverse range of lung tumors with different intrinsic chemosensitivities. PMID- 15805288 TI - Farnesyl transferase inhibitor (R115777)-induced inhibition of STAT3(Tyr705) phosphorylation in human pancreatic cancer cell lines require extracellular signal-regulated kinases. AB - In this study, we report that R115777, a nonpeptidomimetic farnesyl transferase inhibitor, suppresses the growth of human pancreatic adenocarcinoma cell lines and that this growth inhibition is associated with modulation in the phosphorylation levels of signal transducers and activators of transcription 3 (STAT3) and extracellular signal-regulated kinases (ERK). Treatment of cells with R115777 inhibited the tyrosine phosphorylation of STAT3((Tyr705)), while increasing the serine phosphorylation of STAT3((Ser727)). We found the differential phosphorylation of STAT3 was due to an increased and prolonged activation of ERKs. The biological significance of ERK-mediated inhibition of STAT3((Tyr705)) phosphorylation was further assessed by treating the cells with an inhibitor (PD98059) of mitogen-activated protein kinase kinase (MEK) or by transfecting the cells with a vector that expresses constitutively active MEK-1. Expression of constitutively active MEK-1 caused an increase of ERK activity and inhibited STAT3((Tyr705)) phosphorylation. Conversely, inhibition of ERK activity by PD98059 reversed the R115777-induced inhibition of STAT3((Tyr705)) phosphorylation. R115777 also caused the inhibition of the binding of STAT3 to its consensus binding element. An increase in the activation of ERKs either by overexpressing MEK-1 or treatment of cells with R115777 caused an up-regulation in the levels of a cyclin-dependent kinase (cdk) inhibitor, p21(cip1/waf1). These observations suggest that R115777-induced growth inhibition is partly due to the prolonged activation of ERKs that mediates an inhibition of STAT3((Tyr705)) phosphorylation and an increase in the levels of p21(cip1/waf1) in human pancreatic adenocarcinoma cell lines. PMID- 15805289 TI - RNA silencing of checkpoint regulators sensitizes p53-defective prostate cancer cells to chemotherapy while sparing normal cells. AB - p53 is frequently mutated in patients with prostate cancer, especially in those with advanced disease. Therefore, the selective elimination of p53 mutant cells will likely have an impact in the treatment of prostate cancer. Because p53 has important roles in cell cycle checkpoints, it has been anticipated that modulation of checkpoint pathways should sensitize p53-defective cells to chemotherapy while sparing normal cells. To test this idea, we knocked down ataxia telangiectasia mutated (ATM) gene by RNA interference in prostate cancer cell lines and in normal human diploid fibroblasts IMR90. ATM knockdown in p53 defective PC3 prostate cancer cells accelerated their cell cycle transition, increased both E2F activity and proliferating cell nuclear antigen expression, and compromised cell cycle checkpoints, which are normally induced by DNA damage. Consequently, PC3 cells were sensitized to the killing effects of the DNA damaging drug doxorubicin. Combining ATM knockdown with the Chk1 inhibitor UCN-01 further increased doxorubicin sensitivity in these cells. In contrast, the same strategy did not sensitize either IMR90 or LNCaP prostate cancer cells, both of which have normal p53. However, IMR90 and LNCaP cells became more sensitive to doxorubicin or doxorubicin plus UCN-01 when both p53 and ATM functions were suppressed. In addition, knockdown of the G(2) checkpoint regulators ATR and Chk1 also sensitized PC3 cells to doxorubicin and increased the expression of the E2F target gene PCNA. Together, our data support the concept of selective elimination of p53 mutant cells by combining DNA damage with checkpoint inhibitors and suggest a novel mechanistic insight into how such treatment may selectively kill tumor cells. PMID- 15805290 TI - Eradication of tumors from a human colon cancer cell line and from ovarian cancer metastases in immunodeficient mice by a single-chain Ep-CAM-/CD3-bispecific antibody construct. AB - Bispecific T-cell engager (BiTE) are a class of bispecific single-chain antibodies that can very effectively redirect cytotoxic T cells for killing of tumor target cells. Here, we have assessed the in vivo efficacy of one representative, called bscEp-CAMxCD3, with specificity for tumors overexpressing epithelial cell adhesion molecule (Ep-CAM) in human xenograft models. Cells of the human colon carcinoma line SW480 were mixed at a 1:1 ratio with unstimulated human peripheral mononuclear cells, s.c. injected in nonobese diabetes/severe combined immunodeficiency (NOD/SCID) mice, and animals were treated with bscEp CAMxCD3. Five daily i.v. injections of as little as 100 ng per mouse of bscEp CAMxCD3 completely prevented tumor outgrowth when treatment was started at the day of tumor cell inoculation. BscEp-CAMxCD3 was also efficacious when administered up to 8 days after xenograft injection. Established tumors could be eradicated in all animals by five 10 microg doses given between days 8 and 12 after tumor cell inoculation. To test the efficacy of bscEp-CAMxCD3 in a more physiologic model, pieces of primary metastatic tumor tissue from ovarian cancer patients were implanted in NOD/SCID mice. Partial tumor engraftment and growth was observed with four of six patient samples. Treatment of established tumors with daily 5 microg doses led to a significant reduction and, in some cases, eradication of human tumor tissue. These effects obviously relied on the tumor resident T cells reactivated by bscEp-CAMxCD3. Our data show that the class of single-chain bispecific antibodies has very high antitumor efficacy in vivo and can use previously unstimulated T cells at low effector-to-target ratios. PMID- 15805291 TI - A novel ring-substituted diindolylmethane,1,1-bis[3'-(5-methoxyindolyl)]-1-(p-t butylphenyl) methane, inhibits extracellular signal-regulated kinase activation and induces apoptosis in acute myelogenous leukemia. AB - We investigated the antileukemic activity and molecular mechanisms of action of a newly synthesized ring-substituted diindolylmethane derivative, 1,1-bis[3'-(5 methoxyindolyl)]-1-(p-t-butylphenyl) methane (DIM #34), in acute myelogenous leukemia (AML) cells. DIM #34 inhibited AML cell growth via the induction of apoptosis and abrogated clonogenic growth of primary AML samples. Exposure to DIM #34 induced loss of mitochondrial inner transmembrane potential, release of cytochrome c into the cytosol, and caspase activation. Bcl-2-overexpressing, Bax knockout, and caspase-9-deficient cells were partially resistant to cell death, suggesting the involvement of the intrinsic apoptotic pathway. Furthermore, DIM #34 transiently inhibited the phosphorylation and activity of the extracellular signal-regulated kinase and abrogated Bcl-2 phosphorylation. Because other methylene-substituted diindolylmethane analogues have been shown to transactivate the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma), we studied the role of PPARgamma in apoptosis induction. Cotreatment of cells with a selective PPARgamma antagonist or with retinoid X receptor and retinoic acid receptor ligands partially modulated apoptosis when combined with DIM #34, suggesting PPARgamma receptor-dependent and receptor-independent cell death. Together, these findings suggest that diindolylmethanes are a new class of compounds that selectively induce apoptosis in AML cells through the modulation of the extracellular signal-regulated kinase and PPARgamma signaling pathways. PMID- 15805292 TI - RNA interference targeting aurora kinase a suppresses tumor growth and enhances the taxane chemosensitivity in human pancreatic cancer cells. AB - AURKA/STK15/BTAK, the gene encoding Aurora A kinase that is involved in the regulation of centrosomes and segregation of chromosomes, is frequently amplified and overexpressed in various kinds of human cancers, including pancreatic cancer. To address its possibility as a therapeutic target for pancreatic cancer, we employed the RNA interference technique to knockdown AURKA expression and analyzed its phenotypes. We found that the specific knockdown of AURKA in cultured pancreatic cancer cells strongly suppressed in vitro cell growth and in vivo tumorigenicity. The knockdown induced the accumulation of cells in the G(2) M phase and eventual apoptosis. Furthermore, we observed a synergistic enhancement of the cytotoxicity of taxanes, a group of chemotherapeutic agents impairing G(2)-M transition, by the RNA interference-mediated knockdown of AURKA. These results indicate that inhibition of AURKA expression can result in potent antitumor activity and chemosensitizing activity to taxanes in human pancreatic cancer. PMID- 15805293 TI - Targeted delivery of IFNgamma to tumor vessels uncouples antitumor from counterregulatory mechanisms. AB - Because of its immunomodulatory and anticancer activities, IFNgamma has been used as an anticancer drug in several clinical studies, unfortunately with modest results. Attempts to increase the response by increasing the dose or by repeated continuous injection often resulted in lower efficacy, likely due to counterregulatory effects. We show here that targeted delivery of low doses of IFNgamma to CD13, a marker of angiogenic vessels, can overcome major counterregulatory mechanisms and delay tumor growth in two murine models that respond poorly to IFNgamma. Tumor vascular targeting was achieved by coupling IFNgamma to GCNGRC, a CD13 ligand, by genetic engineering technology. The dose response curve was bell-shaped. Maximal effects were induced with a dose of 0.005 microg/kg, about 500-fold lower than the dose used in patients. Nontargeted IFNgamma induced little or no effects over a range of 0.003 to 250 microg/kg. Studies on the mechanism of action showed that low doses of targeted IFNgamma could activate tumor necrosis factor (TNF)-dependent antitumor mechanisms, whereas high doses of either targeted or nontargeted IFNgamma induced soluble TNF receptor shedding in circulation, a known counterregulatory mechanism of TNF activity. These findings suggest that antitumor activity and counterregulatory mechanisms could be uncoupled by tumor vascular targeting with extremely low doses of IFNgamma. PMID- 15805294 TI - Altered IFNgamma signaling and preserved susceptibility to activated natural killer cell-mediated lysis of BCR/ABL targets. AB - Previous studies have shown that BCR/ABL oncogene, the molecular counterpart of the Ph1 chromosome, could represent a privileged target to natural killer (NK) cells. In the present study, we showed that activated peripheral NK cells killed high-level BCR/ABL transfectant UT-7/9 derived from the pluripotent hematopoietic cell line UT-7 with a high efficiency. To further define the mechanisms controlling BCR/ABL target susceptibility to NK-mediated lysis, we studied the effect of IFNgamma, a key cytokine secreted by activated NK cells, on the lysis of these targets. Treatment of UT-7, UT-7/neo, and low BCR/ABL transfectant UT 7/E8 cells with IFNgamma resulted in a dramatic induction of human leukocyte antigen class I (HLA-I) molecules and subsequently in their reduced susceptibility to NK-mediated cytolysis likely as a consequence of inhibitory NK receptors engagement. In contrast, such treatment neither affected HLA-I expression on transfectants expressing high level of BCR/ABL (UT-7/9) nor modulated their lysis by NK cells. Our data further show that the high-level BCR/ABL in UT-7/9 cells display an altered IFNgamma signaling, as evidenced by a decrease in IFN regulatory factor-1 (IRF-1) and signal transducers and activators of transcription (STAT) 1 induction and activation in response to IFNgamma, whereas this pathway is normal in UT-7 and UT-7/E8 cells. A decreased HLA-I induction and nuclear phospho-STAT1 nuclear translocation were also observed in blasts from most chronic myelogenous leukemia patients in response to IFNgamma. These results outline the crucial role of IFNgamma in the control of target cell susceptibility to lysis by activated NK cells and indicate that the altered response to IFNgamma in BCR/ABL targets may preserve these cells from the cytokine-induced negative regulatory effect on their susceptibility to NK mediated lysis. PMID- 15805295 TI - Up-regulation of inhibitory natural killer receptors CD94/NKG2A with suppressed intracellular perforin expression of tumor-infiltrating CD8+ T lymphocytes in human cervical carcinoma. AB - Inhibitory signals that govern the cytolytic functions of CD8(+) T lymphocytes have been linked to the expression of natural killer cell receptors (NKRs) on CTLs. There is limited knowledge about the induction of inhibitory NKR (iNKR) expression in vivo. Up-regulation of iNKRs has been linked to the modulation of the virus- and/or tumor-specific immune responses in animal models. In the present study, we directly examined the expression of various NKRs on tumor infiltrating lymphocytes (TILs) derived from human cervical cancer. We found that in human cervical cancer, the percentage expression of immunoglobulin-like NKR(+)CD8(+) T lymphocytes were similar in gated CD8(+)-autologous TILs and peripheral blood mononuclear cells. On the contrary, cervical cancer-infiltrating CD8(+) T lymphocytes expressed up-regulated C-type lectin NKRs CD94/NKG2A compared with either peripheral blood CD8(+) T cells or normal cervix infiltrating CD8(+) T lymphocytes. Dual NKR coexpression analyses showed that CD94 and NKG2A were mainly expressed on CD56(-)CD161(-)CD8(+) TILs within the cancer milieu. Immunohistochemical study showed that cervical cancer cells expressed abundant interleukin 15 (IL-15) and transforming growth factor-beta (TGF-beta). In kinetic coculture assay, cervical cancer cells can promote the expression of CD94/NKG2A on CD8(+) T lymphocytes. The cancer-derived effects can be reversed by addition of rIL-15Ralpha/Fc and anti-TGF-beta antibody. Functional analyses illustrated that intracellular perforin expression of CD8(+) T cells was minimal upon up-regulation of CD94/NKG2A. Kinetic cytotoxicity assays showed that up-regulated expressions of CD94/NKG2A restrain CD8(+) T lymphocyte cytotoxicity. Our study strongly indicated that cervical cancer cells could promote the expression of iNKRs via an IL-15- and possibly TGF-beta-mediated mechanism and abrogate the antitumor cytotoxicity of TILs. PMID- 15805296 TI - Sensitivity of undifferentiated, high-TCR density CD8+ cells to methylene groups appended to tumor antigen determines their differentiation or death. AB - CD8(+) cells expressing high numbers of TCR per cell (TCR(hi)) are considered important mediators of antitumor effects. To understand the relationship between TCR density and antigen affinity for TCR in the outcome of stimulation with antigen and differentiation of CTL recognizing tumor antigen, we analyzed perforin induction in ovarian tumor-associated lymphocytes in response to the smallest possible changes in the atomic forces of interaction between antigen and TCR. Stimulating undifferentiated, apoptosis-resistant CD8(+) cells expressing high levels of E75-TCR (TCR(hi)) with variants of the CTL epitope E75, HER-2 (369 377), induced their stepwise differentiation, first to IFN-gamma(+) Perf(-) and to TCR(hi) IFN-gamma(+) Perf(+) cells. Blocking caspase-9 activation at antigen stimulation also enhanced the generation of TCR(hi) Perf(hi) cells, demonstrating that TCR density dictated the pathway of death activated by stimulation with the same agonist. Expansion and differentiation of TCR(hi) Perf(+) CTL required an agonist of optimal CH(2) side chain length, which in this study was equal to two CH(2) groups appended to E75 at the Gly(4) position. Side chains one CH(2) shorter or longer than optimal were either less stimulatory or induced death of TCR(hi) Perf(+) cells. Differentiation of TCR(hi) CD8(+) cells can be finely tuned by synthetic amino acids in the peptide, whose side chains induce small increments in the affinity of the antigen for TCR below the affinity which induce apoptosis. PMID- 15805297 TI - Surrogate tumor antigen vaccination induces tumor-specific immunity and the rejection of spontaneous metastases. AB - The nonimmunogenic 4T1 murine mammary carcinoma model and a model surrogate tumor antigen (sTA) were employed to explore the possibility of inducing tumor-specific immunity through active immunization in the absence of defined tumor-associated antigens. Immunization of naive mice with protein-based sTA resulted in protection from s.c. challenge, with 4T1 modified to express the sTA (4T1.sTA), or from a sTA-expressing unrelated tumor cell line (mKSA). Immunization had no effect on parental 4T1 tumor growth or the formation of parental 4T1 spontaneous lung metastases. Mice that were sTA immunized and successfully rejected 4T1.sTA challenge also rejected a subsequent challenge in the contralateral flank with parental 4T1 and strikingly prevented the formation of spontaneous parental 4T1 lung metastases. The rejection of parental 4T1 seemed to be specific for and associated with unknown 4T1 tumor-associated antigens, because rejection of mKSA did not induce cross-protection against a challenge with parental 4T1. To evaluate the effect of this vaccine approach on established disease, mice were simultaneously challenged on day 0 with 4T1.sTA and parental 4T1 in contralateral flanks and then immunized on days 3, 10, 17, and 24 with sTA protein. Tumor growth and metastasis were delayed in four of five animals, and 20% (2 of 5) of the animals were tumor free at the completion of the experiment. Together, these data suggest that prior vaccination with a sTA followed by inoculation with poorly immunogenic tumor cells modified to express the sTA activates determinant spreading and the induction of systemic tumor immunity resulting in indigenous tumor rejection. PMID- 15805298 TI - Altered immunity accompanies disease progression in a mouse model of prostate dysplasia. AB - Increasing evidence suggests that altered immune function accompanies, and indeed may facilitate, cancer progression. In this study, we sought to determine the nature of, and cellular mechanisms underlying, changes in immune status during disease progression in a transgenic mouse model of prostate dysplasia. Immune cells in the tumor microenvironment, as well as in the secondary lymphoid tissues, displayed altered phenotypes. Although evidence of antitumor immunity was detected, there was a paradoxical decrease in the ability of T cells to proliferate in vitro at later stages of disease progression. Detailed analysis of the draining lumbar lymph nodes revealed an increased frequency and number of CD4(+)CD25(+) T cells and an enhanced production of inhibitory cytokines, which correlated with impaired T-cell function. Functional studies confirmed a role for CD4(+)CD25(+) regulatory T cells in suppressing T-cell proliferation as well as regulating the growth of transplanted prostate tumor cells. In addition, our studies show for the first time that anti-CD25 antibody treatment reduces, but does not prevent, tumor growth in a transgenic mouse model of prostate dysplasia. Taken together, this work provides compelling evidence that prostate tumor progression is accompanied by altered immune function and, moreover, that regulatory T cells play an important role in this process. These studies thus provide the impetus for development of specific and effective strategies to deplete regulatory T cells, or suppress their function, as an alternative or adjunct strategy for reducing tumor growth. PMID- 15805299 TI - Aberrant Stat3 signaling by interleukin-4 in malignant glioma cells: involvement of IL-13Ralpha2. AB - Interleukin (IL)-4 exhibits antitumor activity in rodent experimental gliomas, which is likely mediated by the actions of IL-4 on a variety of immune cells present in and around the tumor masses. Here, we show that IL-4, which activates Stat6 in normal human astrocytes and in a variety of other cells, induces an aberrant activation of Stat3 in glioblastoma multiforme (GBM) cells but not in normal human astrocytes. Previously, we have shown that autocrine IL-6 signaling induces a persistent activation of Stat3. Now, we show that Stat3 is further activated by IL-4 stimulation of GBM cells. Expression of IL-13Ralpha2, a decoy receptor for IL-13 that partly blocks IL-4-mediated activation of Stat6 in GBM cells, up-regulates the activation of Stat3 as shown by a small interfering RNA mediated inhibition of IL-13Ralpha2 expression. In addition, transient expression of the IL-13Ralpha2 transgene in 293T cells increases the IL-4-mediated activation of Stat3 and subsequent expression of Stat3-targeted gene. Coimmunoprecipitation results reveal that IL-13Ralpha2-mediated activation of Stat3 does not require a direct physical interaction between Stat3 and IL 13Ralpha2. Chromatin immunoprecipitation assay employing anti-Stat3 antibody confirms the in vivo binding of activated Stat3 to the promoters of genes that encode antiapoptotic proteins Bcl-2, Bcl-x(L), and Mcl-1. IL-4 significantly up regulates of the steady-state levels of Bcl-2, Bcl-x(L), and Mcl-1 in GBM cells. These results indicate that IL-4/IL-13 receptor-mediated Stat3 signaling may contribute to the pathogenesis of GBM cells by modulating the expression of the Bcl-2 family of antiapoptotic proteins. PMID- 15805300 TI - Anti-inflammatory properties of the novel antitumor agent yondelis (trabectedin): inhibition of macrophage differentiation and cytokine production. AB - Yondelis (Trabectedin) is a novel antitumor agent of marine origin extracted from the tunicate Ecteinascidia turbinata. This original compound is active against several human tumors including sarcoma and ovarian and breast adenocarcinoma, as evidenced in phase II clinical trials in advanced multitreated patients. Yondelis is a DNA minor groove binder that blocks cell cycle and interferes with inducible gene transcription in a selective manner. In this study, we investigated the immunomodulatory properties of Yondelis on leukocytes. Human blood monocytes were highly susceptible in vitro to its cytotoxic effect and underwent apoptosis at pharmacologically relevant concentrations (5 nmol/L), whereas lymphocytes were up to 5-fold less sensitive. Macrophages differentiated in vitro with macrophage colony-stimulating factor and tumor-associated macrophages (TAM), isolated from patients with ovarian cancer, were also susceptible. At subcytotoxic concentrations, Yondelis inhibited the in vitro differentiation of monocytes to macrophages. In tumor-treated patients, drug infusion caused a selective decrease of monocyte counts and of ex vivo macrophage differentiation. The in vitro production of two proinflammatory mediators, CCL2 and IL-6, was markedly reduced by Yondelis in monocytes, macrophages, TAM, and freshly isolated ovarian tumor cells. The chemokine CCL2 is the major determinant of monocyte recruitment at tumor sites, whereas IL-6 is a growth factor for ovarian tumors. In view of the protumor activity of TAM and of the strong association between chronic inflammation and cancer progression, the inhibitory effect of Yondelis on macrophage viability, differentiation, and cytokine production is likely to contribute to the antitumor activity of this agent in inflammation-associated human tumors. PMID- 15805301 TI - Association of CYP1A1 polymorphisms with differential metabolic activation of 17beta-estradiol and estrone. AB - Several epidemiologic studies associate certain CYP1A1 genotypes, alone or in combination, with an increased risk of estrogen-related cancers. To answer the question of whether genotype-dependent activation of estrogens by CYP1A1 could be the underlying mechanism, we studied the hydroxylation activity of the most common allelic variants of human CYP1A1 towards both endogenously occurring estrogens, 17beta-estradiol (E2) and estrone (E1). We expressed and purified CYP1A1.1 (wild-type), CYP1A1.2 (Ile(462)Val), and CYP1A1.4 (Thr(461)Asn) and did enzymatic assays of NADPH-dependent estrogen hydroxylation in reconstituted CYP1A1 systems. All CYP1A1 variants catalyzed the formation of 2-, 4-, 6alpha-, and 15alpha-hydroxylated estrogen metabolites from E2 and E1, yet with varying catalytic efficiency and distinct regiospecificity. Whereas the variant CYP1A1.2 (Ile(462)Val) had a significant higher catalytic activity for all hydroxylation sites and both substrates, it was most pronounced for 2-hydroxylation. Catalytic efficiencies for the formation of the major metabolites, 2-OH-E2 and 2-OH-E1, by CYP1A1.2 were 5.7- and 12-fold higher, respectively, compared with the wild-type enzyme. The catalytic efficiencies for hydroxylations catalyzed by CYP1A1.4 were roughly comparable with those of the wild-type enzyme. Enzyme kinetics showed that the superior activity of CYP1A1.2 (Ile(462)Val) is mainly caused by a higher V(max), whereas K(m) values of all variants were similar. The data suggest that risk of estrogen-induced cancers and cardiovascular diseases might be-at least partially-determined by the CYP1A1 genotype. PMID- 15805302 TI - Genetic polymorphism in cytochrome P450 7A1 and risk of colorectal cancer: the Fukuoka Colorectal Cancer Study. AB - Bile acids have long been implicated in the etiology of colorectal cancer, but epidemiologic evidence remains elusive. Cholesterol 7alpha-hydroxylase (CYP7A1) is the rate-limiting enzyme in the synthesis of bile acids from cholesterol in the liver, and thus may be an important determinant of bile acid production. We examined the association between the CYP7A1 A-203C polymorphism and colorectal cancer. The CYP7A1 A-203C polymorphism was determined by the PCR-RFLP method in 685 incident cases of colorectal cancer and 778 controls randomly selected from a community in the Fukuoka area, Japan. The CC genotype was slightly less frequent in the case group, and the adjusted odds ratio for the CC versus AA genotype was 0.88 (95% confidence interval, 0.65-1.20). In the analysis by subsite of the colorectum, a decreased risk associated with the CYP7A1 CC genotype was observed for proximal colon cancer, but not for either distal colon or rectal cancer. The adjusted odds ratios (95% confidence intervals) of proximal colon cancer for the CC genotype were 0.63 (0.36-1.10) compared with the AA genotype, and 0.59 (0.37 0.96) compared with the AA and AC genotypes combined. A decreased risk of proximal colon cancer in relation to the CC genotype of CYP7A1 A-203C, which probably renders less activity of the enzyme converting cholesterol to bile acids, is new evidence for the role of bile acids in colorectal carcinogenesis. PMID- 15805303 TI - The paradoxical association between Bcl-2 expression and prognosis: does the immune system make the difference? PMID- 15805304 TI - Emotional arousal and enhanced amygdala activity: new evidence for the old perseveration-consolidation hypothesis. PMID- 15805305 TI - What's right with my mouse model? New insights into the molecular and cellular basis of cognition from mouse models of Rubinstein-Taybi Syndrome. PMID- 15805306 TI - Estrogen and learning: strategy over parsimony. PMID- 15805307 TI - Olfactory blocking and odorant similarity in the honeybee. AB - Blocking occurs when previous training with a stimulus A reduces (blocks) subsequent learning about a stimulus B, when A and B are trained in compound. The question of whether blocking exists in olfactory conditioning of proboscis extension reflex (PER) in honeybees is under debate. The last published accounts on blocking in honeybees state that blocking occurs when odors A and B are similar (the "similarity hypothesis"). We have tested this hypothesis using four odors (1-octanol, 1-nonanol, eugenol, and limonene) chosen on the basis of their chemical and physiological similarity (experiment 1). We established a generalization matrix that measured perceptual similarity. Bees in the "block group" were first trained with an odor A and, in the second phase, with the mixture AB. Bees in the "novel group" (control group) were first trained with an odor N and, in the second phase, with the mixture AB. After conditioning, bees in both groups were tested for their response to B. We assayed all 24 possible combinations for the four odors standing for A, B, and N. We found blocking in four cases, augmentation in two cases, and no difference in 18 cases; odor similarity could not account for these results. We also repeated the experiments with those six odor combinations that gave rise to the similarity hypothesis (experiment 2: 1-hexanol, 1-octanol, geraniol) and found augmentation in one and no effect in five cases. Thus, blocking is not a consistent phenomenon, nor does it depend on odor similarity. PMID- 15805308 TI - Lasting increases in basolateral amygdala activity after emotional arousal: implications for facilitated consolidation of emotional memories. AB - Manipulations that reduce or enhance the activity of basolateral amygdala (BLA) neurons in the minutes to hours after training have been shown to respectively impair or facilitate retention on the inhibitory avoidance task. Although this suggests that BLA activity is altered after emotional arousal, such changes have not been directly demonstrated. To test this, we devised a feline analog of the inhibitory avoidance task and recorded BLA unit activity before and after a single inescapable footshock. Single-unit recordings revealed that the firing rate of many BLA neurons gradually increased after the footshock, peaking 30-50 min post-shock and then subsiding to baseline levels 2 h later. During this period of increased activity, the discharges of simultaneously recorded BLA cells were more synchronized than before the shock. Although it was known that pairing innocuous (conditioned stimulus, CS) and noxious stimuli modifies the responsiveness of BLA neurons to the CS, our results constitute the first demonstration that emotional arousal produces lasting increases in the spontaneous firing rates of BLA neurons. We propose that these changes in BLA activity may promote Hebbian interactions between coincident but spatially distributed activity patterns in BLA targets, facilitating the consolidation of emotional memories. PMID- 15805309 TI - A role for prefrontal calcium-sensitive protein phosphatase and kinase activities in working memory. AB - The prefrontal cortex is involved in the integration and interpretation of information for directing thoughts and planning action. Working memory is defined as the active maintenance of information in mind and is thought to lie at the core of many prefrontal functions. Although dopamine and other neurotransmitters have been implicated, the intracellular events activated by their receptors that influence working memory are poorly understood. We demonstrate that working memory involves transient changes in prefrontal G(q/11)-signaling and in calcium dependent intracellular protein phosphatase and kinase activity. Interestingly, inhibition of the calcium activated phosphatase calcineurin impaired, while calcium/calmodulin dependent kinase II (CaMKII) and calcium-dependent protein kinase C (PKC) enhanced, working memory. Our findings suggest that the active maintenance of information required for working memory involves transient changes in the balance of these enzymes' activities. PMID- 15805310 TI - Transgenic mice expressing a truncated form of CREB-binding protein (CBP) exhibit deficits in hippocampal synaptic plasticity and memory storage. AB - Deletions, translocations, or point mutations in the CREB-binding protein (CBP) gene have been associated with Rubinstein-Taybi Syndrome; a human developmental disorder characterized by retarded growth and reduced mental function. To examine the role of CBP in memory, transgenic mice were generated in which the CaMKII alpha promoter drives expression of an inhibitory truncated CBP protein in forebrain neurons. Examination of hippocampal long-term potentiation (LTP), a form of synaptic plasticity thought to underlie memory storage, revealed significantly reduced late-phase LTP induced by dopamine-regulated potentiation in hippocampal slices from CBP transgenic mice. However, four-train induced late phase LTP is normal. Behaviorally, CBP transgenic mice exhibited memory deficits in spatial learning in the Morris water maze and deficits in long-term memory for contextual fear conditioning, two hippocampus-dependent tasks. Together, these results demonstrate that CBP is involved in specific forms of hippocampal synaptic plasticity and hippocampus-dependent long-term memory formation. PMID- 15805311 TI - Acute stress facilitates trace eyeblink conditioning in C57BL/6 male mice and increases the excitability of their CA1 pyramidal neurons. AB - The effects of stress (restraint plus tail shock) on hippocampus-dependent trace eyeblink conditioning and hippocampal excitability were examined in C57BL/6 male mice. The results indicate that the stressor significantly increased the concentration of circulating corticosterone, the amount and rate of learning relative to nonstressed conditioned mice, and the excitability of CA1 hippocampal pyramidal neurons. Behaviorally, there was no effect of the stressor on control mice that received unpaired presentations of the tone and periorbital shock, i.e., neither stressed nor nonstressed control mice showed an increase in conditioned responding that was above baseline levels. Biophysically, the stressor significantly decreased the amplitude of the post-burst afterhyperpolarization (AHP) and decreased spike frequency accommodation relative to cells from nonstressed control mice. The effect was significant for mice that were stressed either 1 h or 24 h earlier. The results suggest that the stressor increases the excitability of hippocampal pyramidal neurons and that the mechanism underlying this increase may contribute to the more rapid acquisition of hippocampally dependent eyeblink conditioning. PMID- 15805312 TI - Expression of the immediate-early gene-encoded protein Egr-1 (zif268) during in vitro classical conditioning. AB - Expression of the immediate-early genes (IEGs) has been shown to be induced by activity-dependent synaptic plasticity or behavioral training and is thought to play an important role in long-term memory. In the present study, we examined the induction and expression of the IEG-encoded protein Egr-1 during an in vitro neural correlate of eyeblink classical conditioning. The results showed that Egr 1 protein expression as determined by immunocytochemistry and Western blot analysis rapidly increased during the early stages of conditioning and remained elevated during the later stages. Further, expression of Egr-1 protein required NMDA receptor activation as it was blocked by bath application of AP-5. These findings suggest that the IEG-encoded proteins such as Egr-1 are activated during relatively simple forms of learning in vertebrates. In this case, Egr-1 may have a functional role in the acquisition phase of conditioning as well as in maintaining expression of conditioned responses. PMID- 15805313 TI - Intrahippocampal muscimol shifts learning strategy in gonadally intact young adult female rats. AB - Learning strategy preferences depend upon circulating estrogen levels, with enhanced hippocampus-sensitive place learning coinciding with elevated estrogen levels. The effects of estrogen on strategy may be mediated by fluctuations in GABAergic function, given that inhibitory tone in the hippocampus is low when estrogen is high. We investigated the effects on learning strategy of intrahippocampal injections of a GABA(A) agonist in gonadally intact female rats. On the day of training, rats received 0.3 microL intrahippocampal infusions of muscimol (0.26 nmol or 2.6 nmol) or saline 20 min prior to training on a T-maze in which place (hippocampus-sensitive) or response (striatum-sensitive) strategies offer effective solutions. Muscimol treatment increased the use of the response strategy in a dose-dependent manner without influencing learning speed, indicating that muscimol modulated strategy and not learning ability. Furthermore, the muscimol-related shift to response strategies varied across the estrous cycle. The results indicate that increasing inhibition in the hippocampus biases rats away from hippocampus-sensitive place learning strategies and toward hippocampus-insensitive response learning strategies without a learning deficit. Furthermore, rats at proestrus demonstrated the most dramatic shift in learning strategy following muscimol treatment compared with control conditions, while rats at estrus demonstrated the most complete bias toward response strategies. The enhanced use of hippocampus-sensitive strategies at proestrus likely results from reduced hippocampal inhibition. PMID- 15805314 TI - A latent consolidation phase in auditory identification learning: time in the awake state is sufficient. AB - Large gains in performance, evolving hours after practice has terminated, were reported in a number of visual and some motor learning tasks, as well as recently in an auditory nonverbal discrimination task. It was proposed that these gains reflect a latent phase of experience-triggered memory consolidation in human skill learning. It is not clear, however, whether and when delayed gains in performance evolve following training in an auditory verbal identification task. Here we show that normal-hearing young adults trained to identify consonant-vowel stimuli in increasing levels of background noise showed significant, robust, delayed gains in performance that became effective not earlier than 4 h post training, with most participants improving at more than 6 h post-training. These gains were retained for over 6 mo. Moreover, although it has been recently argued that time including sleep, rather than time per se, is necessary for the evolution of delayed gains in human perceptual learning, our results show that 12 h post-training in the waking state were as effective as 12 h, including no less than 6 h night's sleep. Altogether, the results indicate, for the first time, the existence of a latent, hours-long, consolidation phase in a human auditory verbal learning task, which occurs even during the awake state. PMID- 15805315 TI - Extinction and renewal of Pavlovian modulation in human sequential Feature Positive discrimination learning. AB - Using a conditioned suppression task, we investigated extinction and renewal of Pavlovian modulation in human sequential Feature Positive (FP) discrimination learning. In Experiment 1, in context a participants were first trained on two FP discriminations, X-->A+/A- and Y-->B+/B-. Extinction treatment was administered in the acquisition context a (aaa group) or in a new context b (aba group), and comprised X-->A- extinction and Y- control trials. Discriminative X-->A/A responding was lost in both groups when tested in the extinction context, but partially recovered in the aba and not in the aaa group when tested in the acquisition context, suggesting extinction and renewal of extinguished modulation. The same was observed for the Y-->B/B control pair, however, questioning whether the loss of discriminative X-->A/A responding represented genuine extinction of modulation. In Experiment 2, including only aba groups, participants were trained in context a on two FP discriminations, X-->A+/A- and Y ->B+/B-, after which the group "Extinction" was exposed to X-->A- extinction trials in context b, whereas the group "Control" was exposed to X- control trials; concurrently, both groups received further Y-->B+/B- training. In the group Control, differential Y-->B/B and X-->A/A responding were acquired and maintained throughout the experiment. In the group Extinction, while Y-->B/B responding was also maintained throughout, differential X-->A/A responding disappeared because of X-->A- extinction treatment when tested in the extinction context b, but partially reappeared when tested in the acquisition context a. This evidences aba-renewal of extinguished modulation. PMID- 15805316 TI - Complement C5 and early oxygen kinetics during murine sepsis. AB - OBJECTIVES: Changes in oxygen consumption (VO(2)) and carbon dioxide production (VCO(2)) are common but poorly understood features of sepsis. The authors studied the role of complement C5 in the development of abnormal oxygen kinetics during sepsis in mice, arguing that as a pro-inflammatory event, complement activation might exacerbate disturbances in oxygen use during abdominal sepsis. METHODS: An open-circuit indirect calorimeter was used to measure VO(2), VCO(2), and respiratory exchange ratio during a seven-day median lethal dose (LD(50)) murine cecal ligation and puncture (CLP) model. RESULTS: CLP produced significant changes in oxygen kinetics within three hours of onset, although when the animals were stratified by seven-day survival, no difference in these abnormalities was seen between the survivors and the nonsurvivors. Genetic deficiency of C5 did not ameliorate the changes in oxygen utilization. Rather, the C5-deficient mice experienced more severe abnormalities in oxygen kinetics and greater mortality. Treating animals with anti-C5a antibodies at the time of injury had little effect on oxygen kinetics, indicating that C5b, rather than C5a, was predominantly acting to protect the mice during the first 24 hours of illness. CONCLUSIONS: These findings indicate that the primary contribution of C5 to oxygen kinetics during sepsis is salutary through the host defense conveyed by generation of C5b, rather than detrimental by worsening oxygen utilization via pro-inflammatory mechanisms. PMID- 15805317 TI - Comparison of oxycodone and hydrocodone for the treatment of acute pain associated with fractures: a double-blind, randomized, controlled trial. AB - BACKGROUND: Previous studies have demonstrated the efficacy of oxycodone and hydrocodone for the treatment of acute pain. However, to the best of the authors' knowledge, no previous reports have compared the efficacies of these commonly prescribed agents. OBJECTIVES: To compare the efficacies of oxycodone and hydrocodone for the treatment of acute pain associated with fractures in emergency department (ED) patients. METHODS: This prospective, double-blind, randomized, controlled trial was conducted at an urban trauma center with an annual census of 65,000. Eligible participants included ED patients over the age of 12 years with fractures who consented to participate. Subjects were randomized to receive either oxycodone (5 mg orally [po]) with acetaminophen, or hydrocodone (5 mg po) with acetaminophen. Measurements included demographic information; pain scores on a verbal numeric rating scale at baseline and at 30 and 60 minutes; vital signs at baseline and at 30 and 60 minutes; and adverse effects. Ninety five-percent confidence intervals (95% CIs) constructed about means and proportions were used to assess differences between the oxycodone and hydrocodone groups in analgesic efficacy and side effects. RESULTS: Seventy-three subjects were randomized to receive oxycodone or hydrocodone. Sixty-seven subjects completed the ED study period (n = 35, oxycodone; n = 32, hydrocodone). There was no difference between the two groups in age, weight, gender, ethnicity, diagnoses, baseline pain scores, or vital signs. Patients in both groups had pain relief from baseline to 30 minutes (oxycodone mean change 3.7, 95% CI = 2.9 to 4.6; hydrocodone mean change 2.5, 95% CI = 1.7 to 3.3), and from baseline to 60 minutes (oxycodone mean change 4.4, 95% CI = 3.2 to 5.6; hydrocodone mean change 3.0, 95% CI = 2.1 to 3.9). There was no difference in pain between the patients treated with oxycodone and hydrocodone at 30 minutes (mean difference between groups -0.6, 95% CI = -1.8 to 0.5) or at 60 minutes (mean difference -0.5, 95% CI = -2.0 to 1.0). There was no difference between the groups in nausea, vomiting, itching, or drowsiness; however, the hydrocodone patients had a higher incidence of constipation (oxycodone 0%, hydrocodone 21%, difference in proportions 21%, 95% CI = 3% to 39% more with hydrocodone). CONCLUSIONS: Treatment with acetaminophen and either oxycodone, 5 mg po, or hydrocodone, 5 mg po, resulted in pain relief among ED patients with acute fractures, and there was no difference between the two agents at 30 and 60 minutes. Adverse effect profiles were similar, with the exception of a higher incidence of subsequent constipation with the use of hydrocodone. These results suggest that oxycodone and hydrocodone have similarly potent analgesic effects in the first hour of treatment for ED patients with acute fractures. PMID- 15805318 TI - Ultrasound with topical anesthetic rapidly decreases pain of intravenous cannulation. AB - OBJECTIVES: Physicians and nurses in the emergency department rarely use topical anesthesia when starting intravenous (IV) lines because of time constraints and lack of data on patients' perception of the pain associated with this procedure. Ultrasound pretreatment of skin increases permeation rates of hydrophobic topical medications, including topical lidocaine. The objective of this study was to demonstrate that ultrasound treatment followed by brief application of topical anesthetic decreases the patients' perception of the pain of IV cannulation. METHODS: This was a randomized, controlled, prospective trial performed in a university hospital emergency department. Eighty-seven consecutive English speaking, subcritically injured or ill adult patients able to give consent who were receiving an IV line as part of their care were enrolled. The intervention was a brief ultrasound treatment using the SonoPrep (Sontra Medical Corp., Franklin, MA) therapeutic ultrasound device (frequency, 53-56 kHz) followed by 5 minutes of 4% liposomal lidocaine cream and standard-care IV cannulation, or standard care alone. Participants rated their pain on a visual analog scale. Researchers assessed the site after IV insertion and 20-36 hours later. The primary outcome was participants' subjective pain score, and the secondary outcome was site skin irritation. RESULTS: The ultrasound group reported significantly less pain (p<0.001), with 80% of treated participants reporting pain scores 120 mg/dL, 57 for neutrophils >75%, 15 for glucose concentration <20 mg/dL, and 20 for glucose concentration >120 mg/dL. CONCLUSIONS: When markedly abnormal, results of CSF total protein concentration, glucose concentration, and percent neutrophils have value for diagnosing acute bacterial meningitis, even among children with a low WBC count in CSF. PMID- 15805321 TI - The prevalence and perceived health consequences of hunger in emergency department patient populations. AB - OBJECTIVES: Hunger is associated with chronic poor health and may force patients to make choices that acutely impact their health. This study was conducted to determine the prevalence of hunger, some of the choices it forces, and its perceived health effects among emergency department (ED) patients. METHODS: A validated survey about hunger, choosing between food and medicine, and perceived adverse health outcomes was administered in English, Spanish, and Somali to non critically ill ED patients or their parents. Demographics, rates of hunger, numbers of yearly ED visits, and decisions between buying commodities or medications were assessed. Relationships between variables were summarized with odds ratios (ORs), 95% confidence intervals, and logistic regression. RESULTS: Of 930 respondents, 23.7% screened positive for hunger and 17.6% bought food instead of medications. Among the hunger-positive respondents, 44% believed this choice caused illness, and of these, 69% sought ED assessment and 35% believed they were hospitalized because of this choice. Predictors of hunger were low income (OR = 3.3), illicit drug use (OR = 5.5), chronic illness (OR = 2.1), Hispanic ethnicity (OR = 1.9), and lack of insurance (OR = 1.8). Buying food instead of medicine was predicted by low income (OR = 2.5), chronic illness (OR = 2.4), and lack of insurance (OR = 3.4). Predictors of illness were chronic illness (OR = 1.8), lack of insurance (OR = 2.8), African American race (OR = 2.0), and "poor" health (OR = 1.9). CONCLUSIONS: These ED patients have high rates of hunger and often choose to buy food instead of medications. Many believe that this choice results in ED visits and hospitalizations. Socioeconomic factors may predict hunger and subsequent ill health. PMID- 15805322 TI - Is the Charlson Comorbidity Index useful for predicting trauma outcomes? AB - BACKGROUND: Inclusion of a measure of comorbidity in trauma scoring has been suggested due to the potential for preexisting conditions to impact on patient outcomes, but studies have reported varied results. The Charlson Comorbidity Index (CCI) includes 19 diseases weighted on the basis of their association with mortality, and can be extrapolated from International Classification of Diseases, Ninth Revision (ICD-9) codes for administrative databases. OBJECTIVES: To evaluate the CCI as a predictor of trauma outcome. METHODS: Major trauma patient data from the Victorian State Trauma Registry (VSTR) were used to evaluate the CCI (n = 2,819). The CCI was scored from ICD-10 codes through modification of a previous method of mapping ICD-9 codes to the CCI. Logistic regression was used to determine the association between the CCI and mortality, the effect of adding the CCI to the Trauma and Injury Severity Score (TRISS) methodology, and the impact of adding the CCI to a modification of the TRISS methodology. Model performance was assessed through discrimination and calibration. RESULTS: The CCI was associated with death (p < 0.001), but adding the CCI to TRISS [area under the receiver-operating characteristic curve (AUC) 0.86; 95% CI = 0.84 to 0.88] did not result in improved discrimination over TRISS alone (AUC 0.83; 95% CI = 0.81 to 0.86). Modifying TRISS methodology, with age left as a continuous variable, performed better than the original TRISS (AUC 0.91; 95% CI = 0.89 to 0.92), but the addition of the CCI did not further improve this model (AUC 0.91; 95% CI = 0.89 to 0.92). CONCLUSIONS: While the CCI can be extrapolated from ICD codes and provides a measure of comorbid condition severity and was associated with mortality, addition of the CCI to prediction models did not result in a substantial improvement in performance. PMID- 15805323 TI - Coping responses of emergency physicians and nurses to the 2003 severe acute respiratory syndrome outbreak. AB - OBJECTIVES: During the 2003 severe acute respiratory syndrome (SARS) outbreak, health care workers (HCWs) experienced unusual stressors. The study hospital introduced psychosocial interventions to help HCWs. This study aimed to examine the coping strategies adopted by the emergency department (ED) HCWs who cared for the SARS patients. METHODS: In November 2003, a self-administered questionnaire of physicians and nurses was conducted in the hospital ED that is the national SARS screening center in Singapore. Data collected included demographics and responses to these instruments: 1) the Coping Orientation to Problems Experienced (COPE) to assess coping strategies, 2) the Impact of Event Scale (IES) to measure psychological reactions, and 3) the General Health Questionnaire 28 (GHQ 28) to measure psychiatric morbidity. RESULTS: Thirty-eight of 41 (92.7%) physicians and 58 of 83 (69.9%) nurses responded. The respondents reported a preference for problem-focused and emotion-focused coping measures. The physicians chose humor as a coping response significantly more frequently (p < 0.001) than nurses, scoring 9.61/16 (95% CI = 8.52 to 10.69), compared with the nurses' score of 7.05/16 (95% CI = 6.28 to 7.83). The Filipino HCWs turned to religion as a coping response significantly more frequently (p < 0.001) than the non-Filipino HCWs, scoring 14.38/16 (95% CI = 13.33 to 15.42), compared with 9.93/16 (95% CI = 9.00 to 10.87) for the non-Filipinos. Psychiatric morbidity was 17.7% on the IES and 18.8% on the GHQ 28, with the trend for physicians to report lower psychiatric morbidity. CONCLUSIONS: With a supportive hospital environment, ED HCWs chose adaptive coping in response to the outbreak and reported low psychiatric morbidity. Physicians chose humor and Filipinos chose turning to religion as their preferred responses. Psychosocial interventions to help HCWs need to take these preferences into account. PMID- 15805324 TI - Characteristics of cocaine users presenting to an emergency department chest pain observation unit. AB - OBJECTIVES: This report examines the sociodemographic and substance use characteristics, co-occurring psychological status, substance abuse consequences, and prior experiences with substance abuse treatment among patients with cocaine associated chest pain presenting to an emergency department chest pain observation unit. METHODS: This was a consecutive cohort of patients in the emergency department chest pain observation unit aged 18-60 years with low to moderate risk for acute coronary syndrome and recent cocaine use. Responses on standardized and validated instruments were used to examine demographic and clinical characteristics of the sample and to compare patients who met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria for past three-month substance abuse or substance dependence with patients who did not. RESULTS: Of 145 eligible patients identified between June 1, 2002, and February 29, 2004, 86% met criteria for a lifetime DSM-IV substance use disorder and 50% met past three-month criteria. Approximately one half of the total sample reported substantial symptoms of depression. Substance use frequency and consequences, depression, and psychological distress were significantly more severe among those with past three-month substance use diagnoses; however, most sociodemographic characteristics were not associated with substance use diagnoses. Interest in treatment services and treatment history was also significantly associated with the presence of a substance use disorder diagnosis. CONCLUSIONS: Findings regarding diversity in alcohol and drug involvement, current level of psychological functioning, depressive symptomatology, and interest in treatment services provide useful information for designing emergency department-based interventions for this population. PMID- 15805325 TI - Influenza and emergency department utilization by elders. AB - OBJECTIVES: Influenza outbreaks have been associated with worsened emergency department (ED) crowding. We sought to examine the mechanism behind this association. METHODS: A retrospective time series analysis was conducted in Toronto from January 1996 to April 1999. Weekly data on laboratory-confirmed influenza and other respiratory virus cases in the community and visits to all city EDs (n = 20) were obtained. In longitudinal analyses, we determined the association between influenza and changes in ED utilization by younger and older patients with specific diagnoses grouped as major influenza related (MIR) and upper respiratory infection (URI). Time trends in psychiatric visits and their relationship to influenza were used as a control group. RESULTS: A mean of 11,075 ED visits occurred weekly (SD = 698; average age, 39.9 years; 51% women). Four influenza seasons occurred, with weekly incident case counts ranging from 0 to 236; there were a total of 81 weeks with zero new cases between seasons. In multivariable analyses, every ten new cases of influenza active in the community was associated with a 1.5% (95% confidence interval = 1.2 to 1.8) and 1.2% (95% confidence interval = 0.6 to 1.8) absolute increase in the proportion of ED patients who were elders with MIR conditions and URIs, respectively. Influenza was not significantly associated with ED utilization by younger patients; other respiratory viruses were not significantly associated with ED utilization for any patient group. CONCLUSIONS: Influenza season is associated with increased ED utilization by patients aged 65 years and older, most of whom have major respiratory illnesses and may require hospital admission. No association was seen between influenza and utilization by younger patients. Efforts to reduce the impact of influenza seasons on EDs should focus on elders. PMID- 15805326 TI - Twenty years of emergency medical services for children--a cause for celebration and a call for action. PMID- 15805327 TI - Thrombolysis for acute stroke: the incontrovertible, the controvertible, and the uncertain. PMID- 15805328 TI - A blueprint for a sepsis protocol. AB - Despite numerous advances in medicine, sepsis remains an unconquered challenge. Although outcomes have improved slightly over decades, the unacceptably high mortality rate of 30%-50% for severe sepsis and septic shock continues. However, after years of unsuccessful clinical trials, several investigations over the last few years have reported survival benefit in the treatment of sepsis. Physicians now have several proven therapies to treat sepsis, but have yet to implement them on a widespread, systematic basis. This led 11 international professional societies spanning multiple specialties and continents to come together to create the Surviving Sepsis Campaign. The product of their work is an international effort organized to improve care of patients with sepsis and includes consensus, evidence-based guidelines for care that improves survival in septic patients, and an action plan for change. Given the clear role of early identification and treatment in stopping the sepsis cascade, therapy must start early in the emergency department (ED) and continue throughout the hospital course. The first of the recommendations by the Surviving Sepsis Campaign is the aggressive resuscitation strategy of early goal-directed therapy (EGDT). EGDT is reported to reduce absolute mortality by a staggering 16%. The use of recombinant activated protein C was demonstrated to confer a 6% absolute survival benefit. Steroid supplementation in adrenal insufficiency produced a 10% benefit. Additionally, early and appropriate use of antibiotics remains a cornerstone of therapy. Although no randomized trial will be performed, the effects are undisputed. Finally, although predominantly intensive care unit therapies, tight glucose control and low-tidal-volume ventilation strategies have also led to improved survival. Armed with these new therapies, the medical community must rise to this call to action. Clinicians must change the approach to this disease, as well as the way the septic patient is viewed. Although complex and challenging, these therapies must be brought to the patient's bedside. We propose and describe the Multiple Urgent Sepsis Therapies (MUST) protocol as a practical way to implement a comprehensive treatment plan using available evidence-based therapies. PMID- 15805329 TI - Advanced statistics: bootstrapping confidence intervals for statistics with "difficult" distributions. AB - The use of confidence intervals in reporting results of research has increased dramatically and is now required or highly recommended by editors of many scientific journals. Many resources describe methods for computing confidence intervals for statistics with mathematically simple distributions. Computing confidence intervals for descriptive statistics with distributions that are difficult to represent mathematically is more challenging. The bootstrap is a computationally intensive statistical technique that allows the researcher to make inferences from data without making strong distributional assumptions about the data or the statistic being calculated. This allows the researcher to estimate confidence intervals for statistics that do not have simple sampling distributions (e.g., the median). The purposes of this article are to describe the concept of bootstrapping, to demonstrate how to estimate confidence intervals for the median and the Spearman rank correlation coefficient for non-normally distributed data from a recent clinical study using two commonly used statistical software packages (SAS and Stata), and to discuss specific limitations of the bootstrap. PMID- 15805330 TI - Prevalence of traumatic hip and pelvic fractures in patients with suspected hip fracture and negative initial standard radiographs--a study of emergency department patients. AB - OBJECTIVES: To determine the prevalence of hip and pelvic fractures in emergency department (ED) patients with hip pain and negative standard initial radiographs. METHODS: This was a retrospective, cohort study at an academic, community-based ED. ED patients presenting during a one-year period with hip pain for whom a plain-film radiograph was obtained were included. Eligible patients were identified by query of electronic records. Plain radiographs and magnetic resonance images (MRIs) were ordered at the discretion of the treating physician. Initial plain radiographs were read at the point of care by board-certified radiologists. MRI images were reviewed by radiologists with fellowship training in musculoskeletal imaging. Structured follow-up at more than one month postvisit was conducted to rule out a subsequent diagnosis of hip fracture. Ninety-five percent confidence intervals (95% CIs) and kappa (kappa) were calculated as appropriate. RESULTS: Seven hundred sixty-four of 895 patients (85.3%) had follow up completed (study group). Within the study group, 219 patients (29%) had evidence of fracture on initial radiographs. Of the 545 patients with negative initial radiographs, 62 patients (11.4%) underwent hip MRI during the ED visit. MRI identified 24 additional patients with hip fractures. Interobserver agreement for the presence of fracture on MRI was very good (kappa = 0.847). For patients with negative initial plain radiographs who did not have a hip MRI, follow-up did not identify any of these patients as having a subsequent diagnosis of fracture. Thus, 24 of 545 (4.4%; 95% CI = 3.0% to 6.5%) patients with negative initial plain radiographs had a hip fracture. CONCLUSIONS: In this cohort of ED patients with symptoms suspicious for hip fracture who had negative standard radiographs, the authors found that 4.4% were subsequently diagnosed as having fracture. Further studies are warranted to identify characteristics of patients requiring advanced hip imaging studies. PMID- 15805331 TI - Bladder ultrasound increases catheterization success in pediatric patients. AB - OBJECTIVES: To determine whether volumetric bladder ultrasound (VBUS) determinations improve the rate of successful pediatric catheterizations and caregiver satisfaction. METHODS: This randomized controlled trial was conducted at an urban academic emergency department. Patients younger than 36 months requiring diagnostic urine samples were randomized into one of two groups: VBUS or conventional catheterization (CC). Patients were excluded if they were critically ill or had genitourinary abnormalities. VBUS catheterizations were postponed for 30 minutes if the transverse bladder diameter was <2 cm. Data included demographics, number of catheterizations required for success (>/=2.5 mL urine) (SucC), and the number of postponed catheterizations. Caregiver satisfaction scores using a ten-point Likert scale (1 = poor, 10 = excellent) were obtained for time to SucC, physical handling of child (PhysH), and overall satisfaction. Descriptive statistics, chi-square tests, t-tests, and correlation coefficients were used where appropriate. RESULTS: Of 64 patients, 33 underwent VBUS-guided catheterization. The groups did not differ regarding gender (39% male) and age (mean age: CC, 9.4 months, SD = 7.8; VBUS, 7.7 months, SD = 5.5; p = 0.33). SucC occurred in 94% (VBUS) versus 68% (CC) of patients (p = 0.007). The two groups did not differ on any of the caregiver satisfaction indices (time to SucC: VBUS = 8.9 vs. CC = 8.1; PhysH: VBUS = 9.3 vs. CC = 8.6; overall satisfaction: VBUS = 9.3 vs. CC = 8.5; all p > 0.15). CONCLUSIONS: Although there was no significant difference in caregiver satisfaction between the VBUS and CC groups, implementation of VBUS greatly improved the success rates of pediatric emergency department catheterizations. This improvement is achieved with a rapid and easily mastered VBUS technique. PMID- 15805332 TI - Procedure documentation in emergency medicine residencies: a time of change. AB - OBJECTIVES: The purpose of this study was to identify the methods of procedure documentation (PD) used by emergency medicine residency programs and to ascertain the number of programs that are transitioning to a more advanced system. METHODS: All 122 ACGME-approved allopathic emergency medicine programs were contacted by telephone in December 2001. Survey information was obtained from the program director, an attending physician, a resident, or the residency coordinator. RESULTS: The response rate was 92.6%. Seventeen programs (15%) reported using multiple methods of PD, with only 8% utilizing a formal database. Fifty-five percent reported that PD was manual. One third of all programs utilized a Web based system for PD, while 13% required the use of personal digital assistants (PDAs). Nearly one fifth of programs stated they were changing to another form of PD, with the majority of those changing to a PDA format. Fifteen percent of programs purchased PDAs for their residents, and a similar proportion reported that the PDA was used by "most or all" of their residents to document procedures. Nearly four times as many programs (64%) reported that "most or all" of their residents utilized PDAs for clinical purposes. CONCLUSIONS: PDAs are used by a majority of residents for clinical purposes, although fewer utilize this resource for PD. Although most emergency medicine residency programs still utilize a manual system for PD, many programs are in transition to a more technologically advanced method. PMID- 15805333 TI - A multicenter study of complementary and alternative medicine usage among ED patients. AB - OBJECTIVES: To estimate the prevalence of complementary and alternative medicine (CAM) usage among emergency department (ED) patients. METHODS: This study was a cross-sectional study in four Boston-area EDs. For two 24-hour periods, the authors interviewed consecutive patients age 18 and older about whether they had ever used or had recently (within the past year) used CAM "remedies" (e.g., herbal treatments) or "therapies" (e.g., chiropractic). The authors also asked about the patient's disclosure of CAM usage to allopathic health care providers. RESULTS: Of 978 patients, 752 patients were eligible. Among these, 539 (72%) were interviewed. Of these patients, ever having used CAM was reported by 57% (95% CI = 52% to 61%), and 37% (95% CI = 33% to 41%) reported use in the past year. In a multivariate model of recent CAM usage, independent predictors were younger age, higher education level, and chronic pain. Patients who spoke Spanish as their primary language were less likely to report CAM usage. Disclosure of CAM usage to allopathic providers was low, yet >80% reported that they would be comfortable discussing their CAM usage with allopathic providers. CONCLUSIONS: CAM usage was common among ED patients enrolled in our study, despite underreporting to ED staff. Given the growing evidence of benefits and hazards from popular CAM remedies and therapies, ED staff should consider raising this health issue with their patients. PMID- 15805334 TI - Syncope and NAMCS. PMID- 15805335 TI - Inadequate exposure to patients in the pediatric emergency department. PMID- 15805336 TI - Characteristics of infants with severe retinopathy of prematurity in countries with low, moderate, and high levels of development: implications for screening programs. AB - OBJECTIVE: Retinopathy of prematurity (ROP) is a potentially avoidable cause of blindness in children. The proportion of blindness as a result of ROP varies greatly among countries depending on their level of development, being influenced by the availability of neonatal care, neonatal outcomes, and whether effective screening and treatment programs are in place. The objective of this study was to compare characteristics of premature infants who developed severe ROP between 1996 and 2002 in highly developed countries with less developed countries. METHODS: This was an observational study. A questionnaire was completed by ophthalmologists in countries with low, moderate, and high development rankings (3 highly developed countries and from 10 less well-developed countries) who screen for ROP in which they supplied birth weights and gestational ages (GAs) of infants who were treated for threshold ROP or identified with more advanced stages of the disease. Birth weights and GAs of infants with severe ROP were measured. RESULTS: The mean birth weights of infants from highly developed countries ranged from 737 to 763 g compared with values ranging from 903 to 1527 g in less developed countries. Mean GAs of infants from highly developed countries ranged from 25.3 to 25.6 weeks compared with 26.3 to 33.5 weeks in less developed countries. A total of 13.0% of 1091 infants from poorly developed countries exceeded United Kingdom screening criteria; 3.6% exceeded a criteria of <34 weeks' GA and/or <1750 g birth weight. CONCLUSIONS: These findings suggest that larger, more mature infants are developing severe ROP in countries with low/moderate levels of development compared with highly developed countries. ROP screening programs need to use criteria that are appropriate for their local population. PMID- 15805337 TI - Phototherapy-mediated syndrome of inappropriate secretion of antidiuretic hormone in an in utero selective serotonin reuptake inhibitor-exposed newborn infant. AB - Although selective serotonin reuptake inhibitors (SSRIs) have gained wide acceptance in the off-label treatment of mental disorders in pregnant women, there seems to be an increased risk for serotonergic adverse effects in newborn infants who are exposed to SSRIs during late pregnancy. Hyponatremia as a result of the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) is a relatively common serious side effect of the use of SSRIs in (mostly elderly) adults. Severe hyponatremia as a result of an SIADH is proposed here as part of a neonatal serotonin toxicity syndrome in a newborn infant who was exposed prenatally to an SSRI. The definite reversal to normal serum sodium levels after fluid restriction, the lack of any alternative cause for the SIADH, and the positive temporal relation with a high score on a widely used adverse drug reaction probability scale offer solid support for the hypothesis of a causal relationship between the SIADH and the prenatal sertraline exposure in our neonate. Moreover, accumulative data on the acute enhancement of serotonergic transmission by intense illumination led us to hypothesize that phototherapy used to treat hyperbilirubinemia in the newborn infant could have been the ultimate environmental trigger for this proposed new cause of iatrogenic neonatal SIADH. The speculative role of phototherapy as a physical trigger for this drug-related adverse event should be confirmed in other cases by thorough study of the serotonin metabolism, assay of SSRI levels in cord blood, and serial measurement of plasma levels in exposed neonates. As phototherapy is used frequently in jaundiced neonates and an apparently increasing number of infants are born to mothers who take SSRIs, serotonin toxicity in neonates deserves increased attention. PMID- 15805338 TI - A national survey of pediatric critical care resources in the United States. AB - OBJECTIVE: To characterize resources available for the care of critically ill and injured children in the United States. STUDY DESIGN: In January through May 2004, we conducted a cross-sectional survey of medical directors of intensive care facilities for children. RESULTS: Pediatric critical care medical directors from 257 of 337 eligible hospitals responded to the survey (response rate: 76%). The median number of beds was 12 (interquartile range: 8-17 beds), with a median of 58 admissions per PICU bed (interquartile range: 44-70 admissions per PICU bed) in 2003. The median numbers of admissions per PICU bed were not statistically different among PICUs of different sizes. Fewer than 6% of hospitals shared PICU space with space for critically ill adults. The smallest units (1-6 beds) had higher physician and nurse staffing ratios per PICU bed. Advanced therapeutic technology, particularly renal replacement and inhaled nitric oxide therapy, was significantly more likely to be available in larger PICUs (> or =7 beds). CONCLUSIONS: PICUs with the fewest beds had higher physician and nurse staffing ratios per PICU bed and lower resource capacity for high-intensity renal and respiratory therapy. The impact of PICU resource availability on referral patterns and outcomes of pediatric critical illnesses warrants additional study. PMID- 15805339 TI - Infection control policies and hospital-associated infections among surgical patients: variability and associations in a multicenter pediatric setting. AB - BACKGROUND: Hospital-associated infections are an important cause of patient morbidity and death. Little is known about the variability of infection rates and infection control practices among pediatric hospitals. METHODS: This cross sectional study was performed with the Pediatric Health Information System database, which includes demographic and diagnostic data for 35 freestanding, noncompeting, children's hospitals, and with data from a survey of the hospitals, which yielded additional information on infection control policies and practices. Patients undergoing elective surgical procedures were included in this study. RESULTS: Of the 35 eligible hospitals, 31 (89%) chose to participate in the survey component of this study. A total of 48278 patients met the inclusion criteria for the study; 2.3% of these patients had respiratory infections and 0.8% had gastrointestinal infections. The frequency of patients diagnosed with respiratory or gastrointestinal infections varied considerably among the hospitals and ranged from <1% to 6%. Certain infection control processes also varied among the hospitals during the study period. For instance, of the 31 hospitals, 12 monitored hand hygiene, 19 had administrative support of hand hygiene, and 16 had alcohol hand gel present for the entire study period. The presence of alcohol hand gel for the entire study was strongly and independently associated with lower odds of gastrointestinal infections (adjusted odds ratio: 0.64; 95% confidence interval: 0.49-0.85). CONCLUSIONS: Hospitals should support the use of alcohol hand gel, with the aim of decreasing hospital-associated infection rates. PMID- 15805340 TI - Rapid, needle-free delivery of lidocaine for reducing the pain of venipuncture among pediatric subjects. AB - OBJECTIVES: The purpose of this study was to determine the optimal configuration of an investigational, single-use, needle-free, drug system (ALGRX 3268) that delivers powdered lidocaine into the epidermis for the rapid production of local anesthesia among pediatric subjects undergoing venipuncture. METHODS: Children 3 to 18 years of age were randomly allocated to receive 1 of 3 treatments, ie, (1) placebo, (2) a system configured to deliver 0.25 mg of lidocaine, or (3) a system configured to deliver 0.5 mg of lidocaine, at the antecubital fossa 2 to 3 minutes before venipuncture. Three age groups were included, ie, 3 to 7 years, 8 to 12 years, and 13 to 18 years. Two sets of pain rating scales were used, the Faces Pain Scale-Revised for the youngest age stratum and a visual analog scale for the oldest age stratum. Children in the middle age stratum used both scales. RESULTS: One-hundred forty-four subjects completed the study. For all ages combined, there was a statistically significant and clinically meaningful reduction in pain scores for subjects who received 0.5 mg of lidocaine, compared with placebo. The reduction in pain after 0.25 mg of lidocaine did not achieve statistical significance. CONCLUSIONS: Both active configurations were safe and well tolerated by pediatric subjects undergoing venipuncture at the antecubital fossa. ALGRX 3268 at 0.5 mg, administered 2 to 3 minutes before venipuncture, produced significantly lower pain scores, compared with placebo. PMID- 15805341 TI - Correlates of stress fractures among preadolescent and adolescent girls. AB - OBJECTIVE: Although stress fractures are a source of significant morbidity in active populations, particularly among young female athletes, the causes of stress fractures have not been explored among females <17 years of age or in the general population. The purpose of this study was to examine correlates of stress fractures in a large, population-based, national, cohort study of preadolescent and adolescent girls. METHODS: A cross-sectional analysis of data from 5461 girls, 11 to 17 years of age, in the Growing Up Today Study, an ongoing longitudinal study of the children of registered female nurses participating in Nurses' Health Study II, was performed. Mothers self-reported information regarding their children's histories of stress fractures on their 1998 annual questionnaire. Growing Up Today Study participants self-reported their weight and height, menarcheal status, physical activity, dietary intake, and disordered eating habits on annual surveys. RESULTS: In 1998, the mean age of the participants was 13.9 years. Approximately 2.7% of the girls had a history of stress fracture, 3% engaged in disordered eating (using fasting, diet pills, laxatives, or vomiting to control weight), and 16% participated in > or =16 hours per week of moderate to vigorous activity. Age at menarche, z score of BMI in 1998, calcium intake, vitamin D intake, and daily dairy intake were all unrelated to stress fractures after controlling for age. Independent of age and BMI, girls who participated in > or =16 hours per week of activity in 1998 had 1.88 greater odds of a history of stress fracture than did girls who participated in <4 hours per week (95% confidence interval [CI]: 1.18-3.30). Girls who participated in > or =16 hours per week of activity were also more likely than their peers to engage in disordered eating (4.6% vs 2.8%); however, disordered eating did not have an independent association with stress fractures (odds ratio [OR]: 1.33; 95% CI: 0.61-2.89). Independent of age and BMI, each hour per week of high-impact activity significantly increased the risk of stress fracture (OR: 1.05; 95% CI: 1.02-1.09). Among the high-impact physical activities, only running (OR: 1.13; 95% CI: 1.05-1.22) and cheerleading/gymnastics (OR: 1.10; 95% CI: 1.01-1.21) were independently associated with greater odds of stress fracture. CONCLUSIONS: These findings suggest that, although activity can be beneficial for bone health, there is a threshold over which the risk of stress fracture increases significantly among adolescent girls. High-impact activities, particularly running, cheerleading, and gymnastics, appear to be higher risk than other activities. Prospective studies are needed to explore the directionality of these relationships, as well as the role of menstrual history. In the meantime, clinicians should remain vigilant in identifying and treating disordered eating and menstrual irregularities among their highly active, young, female patients. PMID- 15805342 TI - Safety and efficacy of the nicotine patch and gum for the treatment of adolescent tobacco addiction. AB - OBJECTIVES: To determine the safety and efficacy of the nicotine patch and gum for adolescents who want to quit smoking. DESIGN: Double-blind, double-dummy, randomized, 3-arm trial with a nicotine patch (21 mg), nicotine gum (2 and 4 mg), or a placebo patch and gum; all participants received cognitive-behavioral group therapy. SETTING: Inner-city, outpatient clinic on the East Coast. Subjects. Thirteen- to 17-year-old adolescents who smoked > or =10 cigarettes per day (CPD), scored > or =5 on the Fagerstrom Test of Nicotine Dependence, and were motivated to quit smoking. Intervention. Twelve weeks of nicotine patch or gum therapy with cognitive-behavioral therapy, with a follow-up visit at 6 months (3 months after the end of treatment). MAIN OUTCOME MEASURES: Safety assessed on the basis of adverse event reports for all 3 groups, prolonged abstinence, assessed through self-report and verified with exhaled carbon monoxide (CO) levels of < or =6 ppm, in intent-to-treat analyses, and smoking reduction (CPD and thiocyanate concentrations) among trial completers. RESULTS: A total of 120 participants were randomized (72% white, 70% female; age: 15.2 +/- 1.33 years; smoking: 18.8 +/- 8.56 CPD; Fagerstrom Test of Nicotine Dependence score: 7.04 +/- 1.29) from 1999 to 2003. Participants started smoking at 11.2 +/- 1.98 years of age and had been smoking daily for 2.66 +/- 1.56 years; 75% had at least 1 current psychiatric diagnosis. Mean compliance across groups was higher for the patch (mean: 78.4 82.8%) than for the gum (mean: 38.5-50.7%). Both the patch and gum were well tolerated, and adverse events were similar to those reported in adult trials. Changes in mean saliva cotinine concentrations throughout treatment were not statistically significant. Intent-to-treat analyses of all randomized participants showed CO-confirmed prolonged abstinence rates of 18% for the active patch group, 6.5% for the active-gum group, and 2.5% for the placebo group; the difference between the active-patch and placebo arms was statistically significant. There was no significant effect of patch versus gum or gum versus placebo on cessation outcomes. Abstinence rates at the 3-month follow-up assessment were sustained but were not significantly associated with treatment group. Mean smoking rates, but not CO or thiocyanate concentrations, decreased significantly in all 3 arms but not as a function of treatment group. CONCLUSIONS: Nicotine patch therapy combined with cognitive-behavioral intervention was effective, compared with placebo, for treatment of tobacco dependence among adolescent smokers. Decreases in the numbers of cigarettes smoked appeared to be offset by compensatory smoking. Additional study of nicotine gum, with enhanced instructional support, is needed to assess its efficacy among adolescent smokers. PMID- 15805343 TI - Is chronic fatigue syndrome a connective tissue disorder? A cross-sectional study in adolescents. AB - OBJECTIVES: To investigate whether constitutional laxity of the connective tissues is more frequently present in adolescents with chronic fatigue syndrome (CFS) than in healthy controls. Increased joint hypermobility in patients with CFS has been previously described, as has lower blood pressure in fatigued individuals, which raises the question of whether constitutional laxity is a possible biological predisposing factor for CFS. DESIGN: Cross-sectional study. PARTICIPANTS: Thirty-two adolescents with CFS (according to the criteria of the Centers for Disease Control and Prevention) referred to a tertiary hospital and 167 healthy controls. METHODS: The 32 adolescents with CFS were examined extensively regarding collagen-related parameters: joint mobility, blood pressure, arterial stiffness and arterial wall thickness, skin extensibility, and degradation products of collagen metabolism. Possible confounding factors (age, gender, height, weight, physical activity, muscle strength, diet, alcohol consumption, and cigarette smoking) were also measured. The results were compared with findings in 167 healthy adolescents who underwent the same examinations. RESULTS: Joint mobility, Beighton score, and collagen biochemistry, all indicators of connective tissue abnormality, were equal for both groups. Systolic blood pressure, however, was remarkably lower in patients with CFS (117.3 vs. 129.7 mm Hg; adjusted difference: -13.5 mm Hg; 95% confidence interval [CI]: 19.1, -7.0). Skin extensibility was higher in adolescents with CFS (mean z score: 0.5 vs. 0.1 SD; adjusted difference: 0.3 SD; 95% CI: 0.1, 0.5). Arterial stiffness, expressed as common carotid distension, was lower in adolescents with CFS, indicating stiffer arteries (670 vs 820 mum; adjusted difference: -110 mum; 95% CI: -220, -10). All analyses were adjusted for age, gender, body mass index, and physical activity. Additionally, arterial stiffness was adjusted for lumen diameter and pulse pressure. CONCLUSIONS: These findings do not consistently point in the same direction of an abnormality in connective tissue. Patients with CFS did have lower blood pressure and more extensible skin but lacked the most important parameter indicating constitutional laxity, ie, joint hypermobility. Moreover, the collagen metabolism measured by crosslinks and hydroxyproline in urine, mainly reflecting bone resorption, was not different. The unexpected finding of stiffer arteries in patients with CFS warrants additional investigation. PMID- 15805344 TI - The utility of safety counseling in a pediatric emergency department. AB - OBJECTIVE: Pediatric injuries have a significant impact on the medical system, costing lives and disabling many survivors. Although injury-prevention measures do exist, they are underutilized. A majority of families do not consistently receive counseling by a primary care provider (PCP). We attempted to demonstrate the efficacy of counseling families who presented to a pediatric emergency department with unrelated medical complaints. METHODS: A self-report questionnaire was administered to assess the home safety of patients 2 weeks to 12 years old presenting to the emergency department. Targeted counseling on areas noted to be unsafe was provided, and a 2-week follow-up telephone call was made to assess the effectiveness of the counseling. Information on previous counseling by a PCP was also collected and analyzed. Logistic regression was performed to determine significance and calculate odds ratios. RESULTS: Thirty-seven percent of caregivers recalled receiving any counseling at a PCP visit. Caregivers who had received prior counseling by a PCP were significantly more likely to have a safe home environment. Patients who were English speaking were significantly more likely to have received safety counseling than their Spanish-speaking counterparts. One hundred fifty families received counseling in the emergency department, and 117 were eligible for follow-up. Of these families, 39% made a positive change in the safety of their child's environment at the 2-week telephone follow-up. CONCLUSIONS: Caregivers receiving counseling by a PCP are more likely to provide a safe home environment for their children. Spanish speaking patients are at particularly high risk of not receiving counseling. Of caregivers reporting unsafe practices during an unrelated emergency-department encounter, targeted counseling made a positive impact on behavior after discharge. PMID- 15805345 TI - Delayed diagnosis of Kawasaki syndrome: an analysis of the problem. AB - OBJECTIVE: Most pediatric providers in Colorado are familiar with Kawasaki syndrome (KS). However, in a recent outbreak, 30% of cases were diagnosed after illness day 10. We hypothesized that these children saw providers who were not familiar with KS, were given antibiotics for other diagnoses that delayed identification, had access-to-care issues, or presented atypically. METHODS: A retrospective chart review of 106 consecutive KS cases seen at the Children's Hospital in Denver during 1994-2000 was conducted. RESULTS: Twenty-five of 106 children (23.6%) were diagnosed after day 10 of illness (delayed-diagnosis group [DDG]), and these 25 cases were compared with 81 cases diagnosed on or before day 10 (early-diagnosis group [EDG]). There were no differences between patients in the DDG and EDG in age, gender, number of visits, specialty of the primary care physician, time to the first medical visit, number of antibiotics received, coronary artery abnormalities, white blood cell count, or erythrocyte sedimentation rate. Patients in the DDG had significantly more days of fever, rash, red eyes, and oral changes. A platelet count of >450000/mm3 occurred more often in the DDG (56%) than the EDG (30%). After additional analysis, patients in the EDG had close clustering of symptom onset in the first few days of illness, but patients in the DDG had onset of symptoms scattered over 9 days. Patients in the DDG were 2.8 times more likely to have coronary artery aneurysms than patients in the EDG (DDG: 24%; EDG: 8.6%). CONCLUSIONS: Diagnosis after the 10th day of illness was not linked to type of medical provider, number of antibiotics received, or number of physician visits. Patients in the DDG exhibited the typical features of KS, but the onset of their symptoms was dispersed over time as opposed to the close clustering of symptoms in the EDG. Because coronary artery aneurysms occurred significantly more often in the patients in the DDG, more education is needed to teach health care providers to have a high index of suspicion for KS in young children presenting with fever/rash illnesses. PMID- 15805346 TI - Have there been changes in children's psychosomatic symptoms? A 10-year comparison from Finland. AB - OBJECTIVES: The aims of the study were to determine whether the prevalence of children's somatic symptoms, such as headache, abdominal pain, other pain, and nausea and vomiting, changed from 1989 to 1999 and to study the similarity of parents' and children's reports of the child's symptoms. Furthermore, the aims were to explore possible comorbidity in somatic symptoms and to investigate the associations between somatic and psychiatric symptoms. METHODS: Two cross sectional, representative samples were compared. All children born in 1981 (1989 sample, n = 985) and 1991 (1999 sample, n = 962) and living in selected school districts in southwest Finland served as study samples. The response rate for the 1989 sample was 95% and that for the 1999 sample was 86%. Both children and parents were asked about the children's somatic symptoms, whereas parents, children, and teachers were asked about psychiatric symptoms. To study psychiatric symptoms, the Children's Depression Inventory and Rutter's parent and teacher scales were used. RESULTS: The prevalence of frequent headaches and abdominal pain increased somewhat from 1989 to 1999. Parents often failed to recognize their children's psychosomatic problems. Child-reported somatic symptoms were associated with conduct and hyperactivity symptoms, in addition to a previously well-documented association with depression. In associations between somatic symptoms and psychiatric symptoms, there were some differences between the 1989 and 1999 samples. CONCLUSIONS: In clinical work, questions about somatic and psychiatric symptoms should also be addressed to children themselves, because parents and teachers do not always recognize children's symptoms. When somatic problems are being evaluated, psychiatric symptoms should be asked about, and vice versa. More research is needed to explore the reasons for the increased prevalence of somatic symptoms and their associations with psychiatric symptoms. PMID- 15805347 TI - Short- and long-term beneficial effects of a combined dietary-behavioral-physical activity intervention for the treatment of childhood obesity. AB - BACKGROUND: Obesity has become the most common pediatric chronic disease in the modern era. Early prevention and treatment of childhood and adolescent obesity is mandated. Surprisingly, however, only a minor fraction of obese children participate in weight reduction interventions, and the longer-term effects of these weight-reduction interventions among children have not been elucidated. OBJECTIVE: To examine prospectively the short- and long-term effects of a 3 month, combined dietary-behavioral-physical activity intervention on anthropometric measures, body composition, dietary and leisure-time habits, fitness, and lipid profiles among obese children. METHODS: In this randomized prospective study, 24 obese subjects completed the 3-month intervention and were compared with 22 obese, age- and gender-matched, control subjects. RESULTS: At 3 months, there were significant differences in changes in body weight (-2.8 +/- 2.3 kg vs 1.2 +/- 2.2 kg), BMI (-1.7 +/- 1.1 kg/m2 vs -0.2 +/- 1.0 kg/m2), body fat percentage (from skinfold tests; -3.3 +/- 2.6% vs 1.4 +/- 4.7%), serum total cholesterol level (-24.6 +/- 15.1 mg/dL vs 0.8 +/- 18.7 mg/dL), low-density lipoprotein cholesterol level (-23.3 +/- 15.2 mg/dL vs -3.7 +/- 17.3 mg/dL), and fitness (215 +/- 107 seconds vs 50 +/- 116 seconds) in the intervention group versus the control group. After a 1-year follow-up period, there were significant differences between the intervention group (n = 20) and the control group (n = 20) in body weight (0.6 +/- 6.0 kg vs 5.3 +/- 2.7 kg), BMI (-1.7 +/- 2.3 kg/m2 vs 0.6 +/- 0.9 kg/m2), and body fat percentage. There was a significant increase in leisure-time physical activity among the intervention participants, compared with a decrease among the control subjects. CONCLUSIONS: Our data demonstrate the short- and longer-term beneficial effects of a combined dietary-behavioral physical activity intervention among obese children. These results highlight the importance of multidisciplinary programs for the treatment of childhood obesity and emphasize their encouraging long-term effects. PMID- 15805348 TI - A pulmonary score for assessing the severity of neonatal chronic lung disease. AB - BACKGROUND: Limited data are available to describe the spectrum of severity of neonatal chronic lung disease. In the multicenter Supplemental Therapeutic Oxygen for Prethreshold Retinopathy of Prematurity trial, all infants had some degree of pulmonary dysfunction, because eligibility required a median oxygen saturation of < or =94% with room air. Infants randomized to the supplemental oxygen group (oxygen saturation target of 96-99%) had more pulmonary morbidity than did those in the conventional group (oxygen saturation target of 88-94%). This prompted the retrospective development of a pulmonary severity score to compare the baseline status of the 2 groups. OBJECTIVES: To describe a pulmonary score that reflects the severity of neonatal lung disease and to evaluate the association of the score and its components with subsequent pulmonary morbidity through 3 months of corrected age. DESIGN AND METHODS: A pulmonary score was developed empirically by a consensus panel of 3 neonatologists and was defined as the fraction of inspired oxygen (Fio2) x (support) + (medications), where Fio2 is the actual or "effective" (for nasal cannula) Fio2; support is 2.5 for a ventilator, 1.5 for nasal continuous positive airway pressure, or 1.0 for nasal cannula or hood oxygen; and medications is 0.20 for systemic steroids for bronchopulmonary dysplasia, 0.10 each for regular diuretics or inhaled steroids, and 0.05 each for methylxanthines or intermittent diuretics. The scores could range from 0.21 to 2.95. Pulmonary morbidity was defined as any of the following occurring from randomization at a mean of 35.4 weeks' postmenstrual age through 3 months of corrected age: death or rehospitalization with a pulmonary cause; an episode of pneumonia/sepsis/exacerbation of chronic lung disease; or continued hospitalization, supplemental oxygen therapy, diuretic treatment, or systemic steroid therapy at 3 months. Between-group differences were tested with the Kruskal-Wallis or chi2 test. RESULTS: Data through death or the 3-month corrected age examination were available for 588 infants. Enrolled infants represented a wide spectrum of severity of chronic lung disease, with baseline pulmonary scores at randomization ranging from 0.21 to 2.6. The median pulmonary score at enrollment did not differ between the conventional and supplemental groups (0.42 and 0.45, respectively). However, higher baseline pulmonary scores were observed for infants who did versus did not develop subsequent pulmonary morbidity (0.48 vs 0.38). The pulmonary score was associated with subsequent pulmonary morbidity. Regression analyses adjusting for Supplemental Therapeutic Oxygen for Prethreshold Retinopathy of Prematurity group assignment, gestational age at birth, race, gender, and postmenstrual age at randomization revealed that the score was a significant independent predictor of subsequent pulmonary morbidity (odds ratio: 7.2; 95% confidence interval: 3.6-14.4). CONCLUSIONS: The pulmonary score, calculated near term, reflects a wide spectrum of bronchopulmonary dysplasia severity and is associated with subsequent pulmonary morbidity through corrected age of 3 months. This simple score could prove useful in clinical and research settings. Validation of the score requires additional study. PMID- 15805349 TI - Growth hormone therapy for children born small for gestational age: height gain is less dose dependent over the long term than over the short term. AB - BACKGROUND: Approximately 3% of children are born small for gestational age (SGA), and approximately 10% of SGA children maintain a small body size throughout childhood and often into adult life. Among short SGA children, growth hormone (GH) therapy increases short-term growth in a dose-dependent manner; experience with long-term therapy is limited. OBJECTIVE: To delineate the dose dependency of long-term height gain among short SGA children receiving GH therapy. METHODS: We performed an epianalysis of the first adult height data for SGA children (n = 28) enrolled in 3 randomized trials comparing the growth promoting efficacy of 2 continuous GH regimens (33 or 67 microg/kg per day for approximately 10 years, starting at approximately 5 years of age); in addition, we performed a meta-analysis of the adult height results published previously and those presented here. RESULTS: Epianalysis outcomes (n = 28) suggested that adult height increased more with a higher-dose regimen than with a lower-dose regimen. In the meta-analysis (n = 82), the higher-dose regimen was found to elicit a long term height gain superior to that achieved with the lower-dose regimen by a mean of 0.4 SD (approximately 1 inch). Children who were shorter at the start of therapy experienced more long-term height gain. CONCLUSIONS: These findings confirm GH therapy as an effective and safe approach to reduce the adult height deficit that short SGA children otherwise face. In addition, the first meta analysis indicated that height gain is less dose dependent over the long term than over the short term, at least within the dose range explored to date. For SGA children whose stature is not extremely short, current data support the use of a GH dose of approximately 33 microg/kg per day from start to adult height, particularly if treatment starts at a young age; shorter children (for example, height below -3 SD) might benefit from an approach in which short-term catch-up growth is achieved with a higher dose (> or =50 microg/kg per day) and long-term growth to adult height is ensured with a GH dose of approximately 33 mug/kg per day. Because GH-induced accelerations of height and weight gain evolve in parallel, the dose tapering from > or =50 microg/kg to approximately 33 microg/kg can be accomplished by simply maintaining the absolute GH dose (in micrograms) while the child gains weight (in kilograms). With this algorithm, more growth responsive children taper their GH dose down to approximately 33 microg/kg per day more quickly. PMID- 15805350 TI - Preventing abusive head trauma among infants and young children: a hospital based, parent education program. AB - OBJECTIVE: Abusive head injuries among infants (shaken infant or shaken impact syndrome) represent a devastating form of child abuse; an effective prevention program that reduces the incidence of abusive head injuries could save both lives and the costs of caring for victims. We wished to determine whether a comprehensive, regional, hospital-based, parent education program, administered at the time of the child's birth, could be successfully implemented and to examine its impact on the incidence of abusive head injuries among infants <36 months of age. METHODS: All hospitals that provide maternity care in an 8-county region of western New York State participated in a comprehensive regional program of parent education about violent infant shaking. The program was administered to parents of all newborn infants before the infant's discharge from the hospital. The hospitals were asked to provide both parents (mothers and, whenever possible, fathers or father figures) with information describing the dangers of violent infant shaking and providing alternative responses to persistent infant crying and to have both parents sign voluntarily a commitment statement (CS) affirming their receipt and understanding of the materials. Program compliance was assessed by documenting the number of CSs signed by parents and returned by participating hospitals. Follow-up telephone interviews were conducted with a randomized 10% subset of parents, 7 months after the child's birth, to assess parents' recall of the information. Finally, the regional incidence of abusive head injuries among infants and children <36 months of age during the program (study group) was contrasted with the incidence during the 6 preceding years (historical control group) and with statewide incidence rates for the Commonwealth of Pennsylvania during the control and study periods, using Poisson regression analyses with a type I error rate of 0.05. RESULTS: During the first 5.5 years of the program, 65,205 CSs were documented, representing 69% of the 94,409 live births in the region during that time; 96% of CSs were signed by mothers and 76% by fathers/father figures. Follow-up telephone surveys 7 months later suggested that >95% of parents remembered having received the information. The incidence of abusive head injuries decreased by 47%, from 41.5 cases per 100,000 live births during the 6-year control period to 22.2 cases per 100000 live births during the 5.5-year study period. No comparable decrease was seen in the Commonwealth of Pennsylvania during the years 1996-2002, which bracketed the control and study periods in western New York State. CONCLUSIONS: A coordinated, hospital-based, parent education program, targeting parents of all newborn infants, can reduce significantly the incidence of abusive head injuries among infants and children <36 months of age. PMID- 15805351 TI - Ensuring accurate knowledge of prematurity outcomes for prenatal counseling. AB - OBJECTIVES: To determine the accuracy of knowledge of different health care providers regarding survival and long-term morbidity rates for very premature infants and to examine whether a focused educational intervention improves the accuracy of this knowledge and influences health care decisions. METHODS: Using hypothetical case scenarios with infants at < or =28 weeks of gestation, we surveyed a variety of caregivers involved in perinatal communication and decision making processes at a tertiary center that provides intensive care for neonates. We asked physicians from the pediatrics and obstetrics services and nurses and nurse practitioners from the NICU and obstetrics ward for their best estimates of survival and major long-term disability rates and for their opinions regarding the appropriateness of resuscitation and life support at each week of gestation of <29 weeks. After the survey, we educated all providers about current data on survival and long-term disability rates for preterm infants and gave them pocket sized cards summarizing this information for reference during prenatal counseling. One month after the educational intervention and complete dissemination of the cards, a questionnaire with questions identical to those in the first survey was mailed to the same individuals. RESULTS: Fifty-one health care providers were involved in the baseline survey. The response rates for the postintervention survey were 100% for physicians (20 of 20 subjects) and nurses (20 of 20 subjects) and 91% (10 of 11 subjects) for the nurse practitioners. In the baseline survey, statistically significant underestimates of survival rates were seen for physicians and nurses at 23 to 28 weeks of gestation and for nurse practitioners at 23 to 27 weeks of gestation. Statistically significant overestimates of disability rates were seen for physicians and nurse practitioners at < or =26 weeks of gestation and for nurses at < or =28 weeks of gestation. After the intervention, respondents demonstrated significant improvements in the accuracy of survival and disability estimates at many, but not all, gestational ages. Although underestimation of survival rates and overestimation of disability rates decreased after the intervention, it persisted to some degree. After the intervention, a larger proportion of physicians (53% vs 21%) and a smaller proportion of nurses (10% vs 37%) were likely to recommend resuscitation for infants born at 23 weeks of gestation. CONCLUSIONS: Physicians, nurses, and nurse practitioners underestimated survival rates and overestimated long-term disability rates for very premature infants. After education, their estimates of survival and long-term disability rates for these infants improved significantly. More accurate estimates of survival and disability rates affected physicians' and nurses' theoretical decision-making regarding the appropriateness of resuscitation at 23 weeks of gestation. PMID- 15805352 TI - Physician reimbursement levels and adherence to American Academy of Pediatrics well-visit and immunization recommendations. AB - BACKGROUND: There is concern that commercial health insurance reimbursement levels for immunizations and well-child visits may not be meeting the delivery and practice overhead costs within some areas of the country. There is also concern that insufficient physician reimbursement levels may negatively affect the quality of children's health care. OBJECTIVE: We examined the relationships between commercial health insurance reimbursement levels to physicians for pediatric services and rates of immunization and well visits for children and adolescents. DESIGN: Quality of care was measured by examining state-level immunization and well-visit rates for 2002, which were obtained from the National Committee for Quality Assurance, Health Plan Employer Data and Information Set (HEDIS). Reimbursement data were obtained from the American Academy of Pediatrics Medical Cost Model. Variations in the child and adolescent HEDIS measures were examined as a function of physician reimbursement levels for pediatric services across states. HEDIS data were available for a total of 32 states. Partial correlations controlled for pediatrician concentration, as collected from the US Bureau of the Census and the American Medical Association Physician Masterfile data. RESULTS: Compliance with HEDIS immunization rates for all recommended vaccines was 60% for children and 24% for adolescents. By excluding the varicella vaccine, these rates increased to 70% for children and 44% for adolescents. Adherence rates for well visits were also higher for infants (60%) and children (59%) than for adolescents (34%). Physician reimbursement levels for pediatric services varied from 16.88 dollars per member per month to 32.06 dollars per member per month across states. Statistically significant positive correlations for reimbursement levels were found for 8 of the 16 HEDIS measures examined. Correlations with reimbursement levels were found for childhood immunizations (r = 0.42), infant well visits (r = 0.44), childhood well visits (r = 0.46), and adolescent well visits (r = 0.42). Reimbursement levels were especially strongly related to the rates of adolescent varicella vaccination (r = 0.53). When partial correlations were examined to control for pediatrician concentration, the correlations were reduced by 0.09 on average, suggesting that pediatrician supply may serve as an intermediary of the reimbursement relationship. CONCLUSIONS: Immunization and well-visit rates for infants, children, and adolescents were positively linked with physician reimbursement rates for those services. Although methodologic limitations suggest caution when interpreting these findings, more attention should be given to physician reimbursement levels as a possible predictor of immunization and well-visit rates as measures of quality of care and to the importance of reimbursement levels for pediatrician recruitment. PMID- 15805353 TI - Lengths of stay and costs associated with children's hospitals. AB - OBJECTIVE: Because of the unique mission of freestanding children's hospitals, higher costs have generally been accepted; however, increasing health care costs and the impetus for outcomes data demand more accountability. For common diagnoses, with respect to quality care indicators, length of stay (LOS), and total charges, we propose to compare freestanding children's hospitals and other hospitals. Our hypothesis is that, for similar diagnoses, freestanding children hospitals will have longer LOSs and higher costs than other hospitals. METHODS: Data were analyzed from the Healthcare Cost and Utilization Project Kids' Inpatient Database 2000. Encounters qualified for evaluation when 1 of the top 3 discharge codes was consistent with pneumonia, gastroenteritis, respiratory syncytial virus, dehydration, or asthma. Our outcomes were LOS and total charges per hospital admission; hospitals were categorized as children's hospitals and nonchildren's hospitals. We adjusted for the following potential confounders: number of diagnoses, insurance information, patient age in years, race of patient, admission source, procedures, teaching status of hospital, and hospital location. Because of the right skew of the outcomes, our primary analyses consisted of robust median regression; to support our final models, we also performed sensitivity analyses. RESULTS: Of 252262 total inpatient encounters, 24322 met the inclusion criteria. There were 3408 encounters from 23 different freestanding children's hospitals and 20914 encounters from 1749 nonchildren's hospitals. Freestanding children's hospitals provided care to a higher risk population with more children transferred from other hospitals, a higher percentage of minorities, increased number of co-diagnoses, and a higher percentage on Medicaid. There was no statistically significant difference in LOS by hospital type. However, there was a significant difference in total costs, with the median cost of an admission at freestanding children's hospitals 1294 dollars more per hospitalization than at nonchildren's hospitals, after adjusting for confounders. CONCLUSION: We found no significant difference in median LOS among freestanding children's hospitals and nonchildren's hospitals, but freestanding children's hospitals had higher total charges per admission, even after adjusting for differences in population characteristics. Additional studies are needed to elucidate whether these increased costs result in better health outcomes or are simply attributable to other characteristics of children's hospitals, in which not all patients may benefit. PMID- 15805354 TI - Oral versus vaginal sex among adolescents: perceptions, attitudes, and behavior. AB - OBJECTIVE: Despite studies indicating that a significant proportion of adolescents are having oral sex, the focus of most empirical studies and intervention efforts concerning adolescent sexuality have focused on vaginal intercourse. This narrow focus has created a void in our understanding of adolescents' perceptions of oral sex. This study is the first to investigate adolescents' perceptions of the health, social, and emotional consequences associated with having oral sex as compared with vaginal sex, as well as whether adolescents view oral sex as more acceptable and more prevalent than vaginal sex. METHODS: Participants were 580 ethnically diverse ninth-grade adolescents (mean age: 14.54; 58% female) who participated in a longitudinal study on the relationship between risk and benefit perceptions and sexual activity. Participants completed a self-administered questionnaire that inquired about their sexual experiences and percent chance of experiencing outcomes from, attitudes toward, and perceived prevalence of oral versus vaginal sex among adolescents. RESULTS: More study participants reported having had oral sex (19.6%) than vaginal sex (13.5%), and more participants intended to have oral sex in the next 6 months (31.5%) than vaginal sex (26.3%). Adolescents evaluated oral sex as significantly less risky than vaginal sex on health, social, and emotional consequences. Adolescents also believed that oral sex is more acceptable than vaginal sex for adolescents their own age in both dating and nondating situations, oral sex is less of a threat to their values and beliefs, and more of their peers will have oral sex than vaginal sex in the near future. CONCLUSIONS: Given that adolescents perceive oral sex as less risky, more prevalent, and more acceptable than vaginal sex, it stands to reason that adolescents are more likely to engage in oral sex. It is important that health care providers and others who work with youths recognize adolescents' views about oral sex and broaden their clinical preventive services to include screening, counseling, and education about oral sex. PMID- 15805355 TI - Illness transmission in the home: a possible role for alcohol-based hand gels. AB - OBJECTIVES: The widespread use of child care has altered the epidemiology of respiratory and gastrointestinal (GI) infection in the community. Our primary objective was to measure transmission of respiratory and GI illnesses among families with children enrolled in child care. We also sought to examine potential predictors of reduced illness transmission in the home in a secondary analysis. METHODS: We performed an observational, prospective cohort study to determine transmission rates for respiratory and GI illnesses within families with at least 1 child between 6 months and 5 years of age enrolled in child care. A survey about family beliefs and practices was mailed at the beginning of the study. Symptom diaries were provided for families to record the timing and duration of respiratory and GI illnesses. To ensure the accuracy of symptom diaries, biweekly telephone calls were performed to review illnesses recorded by participants. Families with > or =4 weeks of data recorded were included in the analysis. Families were recruited from 5 pediatric practices in the metropolitan Boston area. Of 261 families who agreed to participate in the study, 208 were available for analysis. Secondary transmission rates for respiratory and GI illnesses were measured as illnesses per susceptible person-month. RESULTS: We observed 1545 respiratory and 360 GI illnesses in 208 families from November 2000 to May 2001. Of these, 1099 (71%) respiratory and 297 (83%) GI illnesses were considered primary illnesses introduced into the home. The secondary transmission rates for respiratory and GI illnesses were 0.63 and 0.35 illnesses per susceptible person-month, respectively. Only two thirds of respondents correctly believed that contact transmission was important in the spread of colds, and fewer than half believed that it was important in the spread of stomach flus. Twenty-two percent of respondents reported use of alcohol-based hand gels all, most, or some of the time; 33% reported always washing their hands after blowing or wiping a nose. In multivariate models, use of alcohol-based hand gels had a protective effect against respiratory illness transmission in the home. CONCLUSIONS: In homes with young children enrolled in child care, illness transmission to family members occurs frequently. Alcohol-based hand gel use was associated with reduced respiratory illness transmission in the home. PMID- 15805356 TI - Evaluation of noise-induced hearing loss in young people using a web-based survey technique. AB - OBJECTIVE: Many adolescents and young adults consciously expose themselves to loud music for entertainment. We hypothesized that these individuals might not be aware that exposure to loud music could result in hearing loss. Furthermore, we wished to assess the feasibility of a web-based survey to collect health information from this group. METHODS: A 28-question survey was designed to target adolescents and young adults. The survey contained questions about views toward general health issues, including hearing loss, and was presented to random visitors at the MTV web site. RESULTS: In 3 days, 9693 web surveys were completed. Hearing loss was defined on a Likert scale as "a very big problem" by 8% of respondents compared with other health issues: sexually transmitted diseases, 50%; alcohol/drug use, 47%; depression, 44%; smoking, 45%; nutrition and weight issues, 31%; and acne, 18%. Notably, most respondents had experienced tinnitus or hearing impairment attending concerts (61%) and clubs (43%). Only 14% of respondents had used earplugs; however, many could be motivated to try ear protection if they were aware of the potential for permanent hearing loss (66%) or were advised by a medical professional (59%). CONCLUSIONS: A majority of young adults have experienced tinnitus and hearing impairment after exposure to loud music. Fortunately, many of these individuals could be motivated to wear ear protection. This novel web-based survey technique rapidly generated a large database and is a feasible method to obtain health data from this group. PMID- 15805357 TI - Attributable cost of nosocomial primary bloodstream infection in pediatric intensive care unit patients. AB - OBJECTIVE: To determine the attributable cost of nosocomial primary bloodstream infections (BSIs) in PICU patients. METHODS: A prospective cohort study was conducted of the PICU of the St Louis Children's Hospital, a 235-bed academic tertiary care center. All patients who were admitted to the PICU were included unless they met the following exclusion criteria: age >18 years, death within 24 hours of PICU admission, admission to the NICU service. Total and direct medical costs of PICU and hospital stay for patients with and without nosocomial primary BSI were measured. RESULTS: Fifty-seven children developed 65 episodes of primary BSIs during their PICU stay. The rate of BSI in this population was 13.8 per 1000 central venous catheter days. In multiple linear regression analysis, severity of illness as measured by the admission Pediatric Risk of Mortality Score III, congenital heart disease, underlying lung disease, ventilator days, transplant (solid organ and bone marrow), and nosocomial primary BSI were independent predictors of PICU direct costs. The direct cost of PICU admission for patients with nosocomial primary BSI was 45,615 dollars and for the patients without primary BSI was 6396 dollars. CONCLUSIONS: After controlling for age, severity of illness, underlying disease, and ventilator days, we found that the direct cost of PICU admission attributable to nosocomial primary BSI was 39,219 dollars. The prevention of these infections through specific interventions is likely to be cost-effective. PMID- 15805358 TI - Treatment of childhood constipation by primary care physicians: efficacy and predictors of outcome. AB - OBJECTIVE: Childhood constipation accounts for 3% of visits to general pediatric clinics and as many as 30% of visits to pediatric gastroenterologists. The majority of children who experience constipation and whose caregivers seek medical care are seen by primary care physicians such as pediatricians or family physicians. Little is known about how primary care physicians treat childhood constipation or the success of their treatments. With this study, we prospectively examined which treatments primary care physicians prescribe to children who present for the first time with constipation and how effective those treatments are. METHODS: A total of 119 children who were between 2 and 7 years of age (mean: 44.1 +/- 13.6 months) and presented to 26 different primary care physicians (15 pediatricians and 11 family physicians) for the treatment of constipation for the first time participated in this study. Parents completed daily diaries of their child's bowel habits for 2 weeks before starting treatment recommended by their primary care physician and again 2 months after treatment. The prescribed treatment was identified by reviewing office records of the treating physicians. RESULTS: After 2 months of treatment, 44 (37%) of 119 children remained constipated. In the majority (87%) of cases, physicians prescribed some form of laxative or stool softener. The most commonly prescribed laxatives were magnesium hydroxide (77%), senna syrup (23%), mineral oil (8%), and lactulose (8%). In nearly all cases, a specific fixed dose of laxative was recommended; in only 5% of cases were parents instructed clearly to adjust the dose of laxative up or down to get the desired effect. In approximately half of the cases, physicians recommended some sort of dietary intervention. Some form of behavioral intervention was mentioned in the office records of approximately one third of cases; however, in most cases, little detail was provided. In 45% of cases, physicians prescribed disimpaction using oral cathartics, enemas, or suppositories followed by daily laxatives. In 35% of cases, physicians prescribed daily laxatives without any disimpaction procedure. In the remainder, physicians prescribed only dietary changes (5%), the use of intermittent laxatives (9%), or no therapy (7%). Treatment success corresponded to how aggressively the child was treated. Specifically, children who underwent some form of colonic evacuation followed by daily laxative therapy were more likely to have responded to treatment than were those who were treated less aggressively. CONCLUSION: Primary care physicians tend to undertreat childhood constipation. After 2 months of treatment, nearly 40% of constipated children remain symptomatic. PMID- 15805359 TI - Variation in inpatient diagnostic testing and management of bronchiolitis. AB - OBJECTIVES: We know little about the variation in diagnosis and management of bronchiolitis. The objectives of this study were (1) to document variations in treatment and diagnostic approaches, lengths of stay (LOSs), and readmission rates and (2) to determine which potentially modifiable process of care measures are associated with longer LOSs and antibiotic usage. METHODS: We used the Pediatric Health Information System, which includes demographic, diagnostic, and detailed patient-level data on 30 large children's hospitals. We examined infants who were younger than 1 year and hospitalized for bronchiolitis (October 2001 September 2003). Multivariate analysis of variance was used to determine whether the variance in the outcomes was hospital related after controlling for other covariates. Linear regression was used to model predictors of increased LOS. Logistic regression was used to model antibiotic usage. Analyses were stratified by age group (<3 months and 3-11 months). RESULTS: A total of 17397 patients were included in the analysis. The mean LOS was 2.97 days; 72% of patients received chest radiographs, 45% received antibiotics, and 25% received systemic steroids. The mean LOS varied considerably across hospitals (range: 2.40-3.90 days), and hospital remained a significant contributor to LOS variation after controlling for our covariates. Variations in the use of diagnostic tests and medications as well as readmission rates also existed and also remained significant after controlling for covariates. The factors associated with the greatest increases in LOS in the regression analyses included higher severity scores and use of antibiotics, bronchodilators, and corticosteroids. The strongest predictors of antibiotic use in the logistic regression analyses were higher severity scores and receipt of a blood or cerebrospinal fluid culture. Receiving a chest radiograph was a significant predictor of antibiotic use in older but not younger infants. CONCLUSIONS: Considerable, unexplained variation exists in the inpatient management of bronchiolitis. The development of national guidelines and controlled trials of new therapies and different management approaches are indicated. PMID- 15805360 TI - Yield of diagnostic testing in infants who have had an apparent life-threatening event. AB - OBJECTIVE: Many infants who have experienced an apparent life-threatening event (ALTE) seem normal by the time they reach the hospital. Nevertheless, they typically undergo an extensive evaluation to rule out serious underlying conditions. The purpose of the present investigation was to determine the yield of different diagnostic tests in helping to identify the cause of the ALTE. METHODS: We reviewed test results from a consecutive series of infants who were younger than 12 months and admitted to a tertiary care academic medical center between November 1996 and June 1999 after having experienced a sudden breathing irregularity, color change, or alteration in mental status or muscle tone. For each patient, we noted whether a given test was performed, whether the result was positive, and, if so, whether the test contributed to the diagnosis, that is, suggested or helped establish the cause. We also noted whether the initial history and physical examination contributed information that eventually led to the final diagnosis. RESULTS: A total of 243 patients met the enrollment criteria. Of the 3776 tests ordered, 669 (17.7%) were positive and 224 (5.9%) contributed to the diagnosis. Prompted by findings from the initial clinical assessment, the following tests proved useful in patients who had a contributory history and physical examination: blood counts, chemistries, and cultures; cerebrospinal fluid analysis and cultures; metabolic screening; screening for respiratory pathogens; screening for gastroesophageal reflux; chest radiograph; brain neuroimaging; skeletal survey; electroencephalogram; echocardiogram; and pneumogram. In the remaining patients, who had a noncontributory history and physical examination, only the following tests proved useful: screening for gastroesophageal reflux, urine analysis and culture, brain neuroimaging, chest radiograph, pneumogram, and white blood cell count. Broad evaluations for systemic infections, metabolic diseases, and blood chemistry abnormalities were not productive in these patients. CONCLUSIONS: For many tests used in the evaluation of an ALTE, the likelihood of a positive result is low and the likelihood of a contributory result is even lower. Estimates of diagnostic yield derived from the present investigation can help clinicians maximize the productivity and efficiency of their evaluation. PMID- 15805361 TI - Adult health and social outcomes of children who have been in public care: population-based study. AB - OBJECTIVE: To examine adult socioeconomic, educational, social, and health outcomes of being in public care in childhood. METHODS: The 1970 British birth cohort was followed up at 5 (N = 13135), 10 (14875), 16 (11622), and 30 years (11261). Cases were defined as those ever in statutory or voluntary public care at 5, 10, and 16 years. Self-reported adult outcomes were occupation, educational achievement, general health, psychological morbidity, history of homelessness, school exclusion, and convictions. RESULTS: A total of 343 (3.6%) of 9557 had been in public care <17 years. Nonwhite children were more likely to have been in care (odds ratio [OR]: 3.3; 95% confidence interval [CI]: 2.1-5.4). Controlling for socioeconomic status, men with a history of public care were less likely to attain high social class (OR: 0.6; 95% CI: 0.4-0.9) and more likely to have been homeless (OR: 2.0; 95% CI: 1.1-3.8), have a conviction (OR: 2.3; 95% CI: 1.5 3.4), have psychological morbidity (OR: 1.8; 95% CI: 1.1-3.0), and be in poor general health (OR: 1.6; 95% CI: 1.1-2.6). Similar associations were found in women. Men but not women with a history of care were more likely to be unemployed (OR: 2.6; 95% CI: 1.4-5.0) and less likely to attain a higher degree (OR: 0.4; 95% CI: 0.2-0.7). Nonwhite ethnicity was associated with poorer adult outcomes of being in care. CONCLUSIONS: Public care in childhood is associated with adverse adult socioeconomic, educational, legal, and health outcomes in excess of that associated with childhood or adult disadvantage. PMID- 15805362 TI - Importance of catch-up vaccination: experience from a varicella outbreak, Maine, 2002-2003. AB - OBJECTIVE: During December 2002 to January 2003, a varicella outbreak occurred in an elementary school in Maine. Just 1 month before detecting the outbreak, Maine implemented varicella vaccine requirements for child care but did not require vaccination for school entry. We investigated this outbreak to examine reasons for its occurrence, including vaccine failure. METHODS: A self-administered questionnaire was sent to all students' parents to determine student disease status, medical conditions, and vaccination status, which was further confirmed by review of medical records. Parental reporting of chickenpox/varicella that occurred since September 1, 2002, in a student who attended the school was used to define a case. Parents of cases were interviewed by telephone about disease characteristics. Disease severity was classified on the basis of the number of skin lesions and the occurrence of complications. Vaccine effectiveness was calculated by comparing varicella attack rates for any disease, for moderate to severe disease, and for severe disease among vaccinated and unvaccinated students. RESULTS: We obtained complete information for 296 (81%) of 364 students. Varicella vaccine coverage was 74% overall and decreased by grade, from 90% in kindergarten to 60% in third grade. Attack rates increased significantly from 14% in kindergarten to 37% in third grade. Of the 53 varicella cases, 36 (68%) were unvaccinated, 12 (22%) were vaccinated, and 5 (10%) had previous disease history. Vaccine effectiveness was 89% (95% confidence interval [CI]: 79 94%) against disease of any severity, 96% (95% CI: 88-99%) against moderate to severe disease, and 100% (95% CI: undefined) against severe disease. Twenty-two percent of unvaccinated students had severe disease and 1 was hospitalized for a skin infection, whereas none of the vaccinated cases reported severe disease. CONCLUSION: This outbreak was attributable primarily to failure to vaccinate, especially among children in grades 1 through 3. Catch-up vaccination of susceptible older children and adolescents is especially important to prevent accumulation of susceptibility in these groups, in which the natural disease is more severe. School entry requirements will contribute to a more rapid implementation of the existing recommendations for vaccination. PMID- 15805363 TI - Estimated impact of competing policy recommendations for age of first dental visit. AB - OBJECTIVE: To compare levels of dental utilization and untreated dental decay among children aged 1 to 3 years that are likely to occur under 2 potential guidance policies: (1) pediatricians refer all toddlers to dentists for screening (consistent with American Academy of Pediatric Dentistry and the American Dental Association recommendations; DENT), and (2) pediatricians receive training in caries risk assessment, screen toddlers, and refer at-risk children to dentists (consistent with American Academy of Pediatrics recommendations; PED). METHODS: Using decision analysis, we estimated the impact of PED and DENT assuming alternately unlimited dental capacity for Medicaid-insured patients and fixed Medicaid dental capacity. Results With unlimited capacity, if DENT were implemented, then dental utilization is estimated to increase from 27% under the status quo to 65% and untreated decay to decrease from a mean of 0.60 surfaces to 0.52 surfaces per child. If PED were implemented, then dental utilization and untreated decay would decrease from status quo levels to an estimated 11% and 0.47 surfaces, respectively, assuming that diagnostic sensitivity and specificity both equaled 1; they would decrease to 13% and 0.53 surfaces, respectively, if sensitivity equaled 0.76 and specificity equaled 0.95. With fixed capacity, under DENT, untreated decay is estimated to increase to 0.63 surfaces because low-risk private-pay patients would crowd out at-risk Medicaid-insured children, whereas under PED, untreated decay would still be less than under the status quo. CONCLUSIONS: Implementing PED will decrease untreated decay under most plausible scenarios, whereas switching to DENT will increase the burden of disease if Medicaid dental capacity is limited. PMID- 15805364 TI - Helping parents cope with the trauma of premature birth: an evaluation of a trauma-preventive psychological intervention. AB - OBJECTIVE: To ascertain whether a trauma-preventive psychological intervention program for parents of premature infants during hospitalization in a level III NICU may reduce the severity of symptomatic response to the traumatic impact of premature birth. METHODS: Mothers of premature infants were enrolled consecutively in a sequential control group design. Intervention group mothers received a structured psychological intervention in the first days after birth. Each mother could make use of additional psychological support if required and was actively approached at critical times during her infant's NICU stay. Control group mothers did not receive psychological intervention but could ask for counseling by the hospital minister. At discharge, mothers of both groups answered a questionnaire covering key outcome variables (symptoms of traumatization, emotions at discharge, and sample and control variables). RESULTS: At discharge, intervention group mothers (N = 25) showed significantly lower levels of symptomatic response to the traumatic stressor "premature birth" than those in the control group (N = 25; mean overall symptom level 25.2 [SD: 13.9] vs 37.5 [SD: 19.2]). CONCLUSIONS: This intervention program for parents after premature birth, combining early crisis intervention, psychological aid throughout the infant's hospitalization, and intense support at critical times, reduced the symptoms of traumatization relating to premature birth. PMID- 15805365 TI - Effects of race, insurance status, and hospital volume on perforated appendicitis in children. AB - OBJECTIVE: Previous research suggests that perforated appendicitis is more common in Medicaid patients, but the roles of minority race and hospital volume remain largely unstudied. We sought to investigate the association of perforated appendicitis in children with minority race, insurance status, and hospital volume. METHODS: We conducted a retrospective, population-based cohort study of 33184 children who had an International Classification of Diseases, Ninth Revision diagnosis code for acute appendicitis in The Kids' Inpatient Database, a pediatric database from 22 states in 1997. A multivariate logistic regression model was developed to determine patient and hospital characteristics predictive of perforated appendicitis. RESULTS: Of 33184 children with acute appendicitis, 10777 (32.5%) were perforated. In multivariate analysis, black (odds ratio [OR]: 1.24; 95% confidence interval [CI]: 1.10-1.39) and Hispanic (OR: 1.19; 95% CI: 1.10-1.29) children were more likely to have perforated appendicitis than white children. Perforation was also more likely in Medicaid patients (OR: 1.30; 95% CI 1.22-1.39) compared with privately insured children. Annual hospital volume of cases of appendicitis was not significantly associated with perforation in multivariate analysis. CONCLUSIONS: Perforated appendicitis disproportionately affected both children of minority race and children insured by Medicaid. No effect of hospital volume was observed. To reduce this racial disparity, efforts should focus on the causes of delayed diagnosis and the treatment of appendicitis in children of minority race. PMID- 15805366 TI - Neonatal Ventilation With Inhaled Nitric Oxide Versus Ventilatory Support Without Inhaled Nitric Oxide for Preterm Infants With Severe Respiratory Failure: the INNOVO multicentre randomised controlled trial (ISRCTN 17821339). AB - BACKGROUND: Although inhaled nitric oxide (iNO) may be a promising treatment for newborn infants with severe respiratory failure, the results from 3 previous small trials were inconclusive. METHODS: Infants of <34 weeks' gestation, <28 days old, and with severe respiratory failure requiring ventilatory support were randomized to receive or not receive iNO. The study was not blinded. FINDINGS: Recruited were 108 infants (55 allocated to receive iNO and 53 not allocated to receive iNO) from 15 neonatal units in the United Kingdom and Republic of Ireland. Fifty-nine percent (64 of 108) died, and 84% of the survivors (37 of 44) had signs of some impairment or disability, 9 (20%) of them classified as severely disabled. There was no evidence of an effect of iNO on the primary outcomes: death or severe disability at 1 year corrected age (relative risk [RR]: 0.99; 95% confidence interval [CI]: 0.76 to 1.29); death or supplemental oxygen on expected date of delivery (RR: 0.84; 95% CI: 0.68 to 1.02); or death or supplemental oxygen at 36 weeks' postmenstrual age (RR: 0.98; 95% CI: 0.87 to 1.12). There was a trend for infants allocated to the iNO group to spend more time on the ventilator (log rank: 3.6), on supplemental oxygen (log rank: 1.4), and in hospital (log rank: 3.5) than those allocated to receive no iNO. This pattern predominantly reflected the infants who died. Mean total costs at 1 year corrected age were significantly higher in the iNO group, partly because of the costs of the gas but mainly because of the difference in initial hospitalization costs. INTERPRETATION: Evidence of prolongation of intensive care and increased costs of such care, without clear beneficial effects, implies that iNO cannot be recommended for preterm infants with severe hypoxic respiratory failure. PMID- 15805367 TI - Heart rate characteristics and laboratory tests in neonatal sepsis. AB - OBJECTIVE: The evaluation of an infant for suspected sepsis often includes obtaining blood for laboratory tests. The shortcomings of the current practice are that the infant has to appear clinically ill for the diagnosis to be entertained, and the conventional laboratory tests are invasive. We have found that the clinical diagnosis of neonatal sepsis is preceded by abnormal heart rate characteristics (HRC) of reduced variability and transient decelerations, and we have devised a predictive HRC monitoring strategy based on multivariable logistic regression analysis that was developed at one tertiary care NICU and validated at another. We hypothesized that HRC monitoring, which is continuous and noninvasive, might be an adjunct to conventional laboratory tests in the diagnosis of neonatal sepsis. The objective of this study was to test the hypothesis that HRC monitoring adds information to conventional laboratory tests in diagnosing neonatal sepsis. METHODS: We prospectively collected heart rate data in 678 consecutive infants who stayed >7 days in the University of Virginia NICU from July 1999 to July 2003. We prospectively measured HRC and noted 149 episodes of sepsis with positive blood cultures for which data were available in 137. We obtained all laboratory test results for ratio of immature to total neutrophil count, white blood cell count, glucose, platelet count, HCO3, arterial partial pressure of carbon dioxide, and pH. We tested hypotheses using multivariable logistic regression modeling adjusted for repeated measures. RESULTS: We found that the HRC index, which was available 92% of the time, was highly significantly associated with sepsis (receiver-operating characteristic [ROC] area: 0.73). The ratio of immature to total neutrophil count, white blood cell count (available 4%-8% of the time, usually around the time of suspected sepsis), and blood glucose and pH (available 28% and 38% of the time) were also significantly associated with sepsis (ROC area: 0.75). HRC and laboratory values added independent information to each other, and a predictive model using all significant variables had ROC area of 0.82. CONCLUSIONS: HRC monitoring adds independent information to laboratory tests in the diagnosis of culture-positive neonatal sepsis. PMID- 15805368 TI - Risk factors for and outcomes of bloodstream infection caused by extended spectrum beta-lactamase-producing Escherichia coli and Klebsiella species in children. AB - OBJECTIVE: The increasing prevalence of infections caused by extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella species (ESBL-EK) has become a growing concern in the hospitalized patient population. Previous studies on risk factors for infection with ESBL-EK have mainly focused on adult populations, and these findings may not be relevant among the pediatric population that experiences a unique set of health care exposures and underlying conditions. The objective of this study was to define the risk factors and outcomes associated with ESBL-EK bloodstream infections in children. METHODS: We conducted a nested case-control study using data from the Children's Hospital of Philadelphia from May 1, 1999, to September 30, 2003. Eligible patients were identified from the hospital database of microbiology laboratory records. All patients with ESBL-EK bloodstream infections were compared to a random sample of patients with non-ESBL-EK bloodstream infections. Risk factors analyzed included prior antimicrobial use, comorbid conditions, and demographic characteristics. Pulsed-field gel electrophoresis was performed to determine genetic relatedness of the ESBL-EK isolates. RESULTS: Thirty-five cases and 105 control subjects were included in the study. The median age among the cases was 2 years (interquartile range: 0-11), compared with 1 year (interquartile range: 0-8) among control subjects. Patients with ESBL-EK infections were 5.8 times (95% confidence interval: 1.9-17.7) more likely to have had exposure to an extended-spectrum cephalosporin in the 30 days before infection than those with non-ESBL-EK infections. Other independent predictors of ESBL-EK infection were being female, infection with a Klebsiella species, and steroid use in the 30 days before infection. All ESBL-EK isolates were susceptible to carbapenem antibiotics. Pulsed-field gel electrophoresis analysis revealed that the ESBL-EK isolates were polyclonal. Although a substantially higher proportion of children with ESBL-EK died (in-hospital mortality: 36% vs 13%), this difference was not statistically significant. CONCLUSIONS: Receipt of extended-spectrum cephalosporins in the 30 days before infection by an Escherichia coli or Klebsiella species is significantly associated with having an ESBL-EK infection in hospitalized children. Curtailed use of cephalosporins among high-risk groups may reduce the occurrence of ESBL-EK infections. Future studies on identifying high-risk children and investigating the impact of curtailed third-generation cephalosporin use to limit additional emergence of ESBL-EK infections should be undertaken. PMID- 15805369 TI - Use of volume expansion during delivery room resuscitation in near-term and term infants. AB - OBJECTIVE: To characterize use of volume infusion (VI) for infants who are > or =34 weeks' gestational age and receive intensive cardiopulmonary resuscitation (CPR; defined as >1 minute of positive-pressure ventilation and chest compressions, with or without the administration of medications) in the delivery room and are admitted to the NICU. METHODS: A retrospective review of a resuscitation registry between January 1999 and June 2001 was conducted. RESULTS: Of 37,972 infants, 23 received CPR, including 13 with VI. Ten of 13 received VI for persistent bradycardia despite CPR, and only 3 of 13 received VI for suspicion of hypovolemia with poor perfusion. More VI versus no VI infants had Apgar scores < or =2 at 5 and 10 minutes. VI versus no VI infants had lower cord arterial pH, had higher arterial partial pressure of carbon dioxide, had larger base deficit, required longer CPR, and required more epinephrine. On admission to the NICU, VI versus no VI infants had lower blood pressure and larger base deficit over the first 2 hours but did not differ in arterial pH, arterial partial pressure of carbon dioxide, heart rate, mortality, or use of additional VI or buffer. CONCLUSIONS: VI is rarely given for overt hypovolemia and more often for asphyxiated infants who are slow to respond to intensive CPR. Persistent postnatal hypotension in VI infants suggests that other factors, eg, myocardial dysfunction, may be important contributors to lack of response to CPR. PMID- 15805370 TI - Primary and secondary nocturnal enuresis: similarities in presentation. AB - OBJECTIVE: To determine the differences or similarities in the clinical presentation between patients with primary and secondary nocturnal enuresis. METHODS: A total of 170 patients with nocturnal enuresis were assessed at a busy tertiary care pediatric voiding dysfunction clinic at the University of Oklahoma Health Sciences Center. Patients with primary nocturnal enuresis (PNE) were compared with patients with secondary nocturnal enuresis (SNE) for a variety of clinical features, including gender, age when first voiding on their own, age on presentation, infrequent voiding, frequent voiding, urgency, daytime wetting, nocturia, urinary tract infection, constipation, vesicoureteral reflux, attention deficit/hyperactivity disorder, uroflow results, and ultrasound evidence of a postvoid residual. RESULTS: The only significant difference between the patients with PNE and those with SNE was in the prevalence of constipation. Constipation was significantly associated with PNE (74.59% vs 57.54%; odds ratio: 2.17; 95% confidence interval: 1.07-4.41). When adjusted for a history of constipation, the age at which a child began to void on his or her own became statistically significant. Patients with SNE started to void on their own at 2.13 years (SD: 0.61), an average of 0.22 years earlier than those with PNE, who started to void on their own at 2.35 years. CONCLUSIONS: PNE and SNE likely share a common pathogenesis. Symptoms of daytime voiding dysfunction are common in patients with PNE and SNE. Daytime voiding habits might influence how the central nervous system responds at night to a full or contracting bladder. PMID- 15805371 TI - Increasing the screening and counseling of adolescents for risky health behaviors: a primary care intervention. AB - OBJECTIVE: To determine whether a systems intervention for primary care providers resulted in increased preventive screening and counseling of adolescent patients, compared with the usual standard of care. METHODS: The intervention was conducted in 2 outpatient pediatric clinics; 2 other pediatric clinics in the same health maintenance organization served as comparison sites. The intervention was implemented in 2 phases: first, pediatric primary care providers attended a training workshop (N = 37) to increase screening and counseling of adolescents in the areas of tobacco, alcohol, drugs, sexual behavior, and safety (seatbelt and helmet use). Second, screening and charting tools were integrated into the intervention clinics. Providers in the comparison sites (N = 39) continued to provide the usual standard of care to their adolescent patients. Adolescent reports were used to assess changes in provider behavior. After a well visit, 13- to 17-year olds (N = 2628) completed surveys reporting on whether their provider screened and counseled them for risky behavior. RESULTS: Screening and counseling rates increased significantly in each of the 6 areas in the intervention sites, compared with rates of delivery using the usual standard of care. Across the 6 areas combined, the average screening rate increased from 58% to 83%; counseling rates increased from 52% to 78%. There were no significant increases in the comparison sites during the same period. The training component seems to account for most of this increase, with the tools sustaining the effects of the training. CONCLUSIONS: The study offers strong support for an intervention to increase clinicians' delivery of preventive services to a wide age range of adolescent patients. PMID- 15805372 TI - Is noncompliance among adolescent renal transplant recipients inevitable? AB - OBJECTIVE: To evaluate the prevalence of noncompliance and factors that influence poor adherence to immunosuppressive drug regimens among kidney transplant recipients. METHODS: We reviewed immunosuppressive drug compliance in 79 posttransplant patients. Patient self-report and low plasma calcineurin inhibitor levels served as indicators of noncompliance. RESULTS: The prevalence of noncompliance was found to be highest in adolescents who were responsible for their own medications and who underwent cadaveric kidney transplantation (CTx; 45.5%) and lower after living related transplantation (28.6%). There were no documented cases of noncompliance among any recipient of living unrelated (commercial) transplantation. Among 13 noncompliant patients, the first indication of "drug holiday" was low plasma calcineurin inhibitor levels in 11 children. Two additional children presented with acute rejection. In 7 patients, repeated episodes of "drug holidays" led to acute rejection later: 21.4 +/- 13.2 months after the first decrease in plasma calcineurin inhibitor level had been recorded. All 9 patients who experienced acute rejection subsequently developed chronic rejection. In 4 patients, noncompliance did not influence graft function. Psychosocial factors that were associated with noncompliance included insufficient family support, low self-awareness caused by poor cognitive abilities, and denial. CONCLUSIONS: The absence of cases of noncompliance in adolescents who underwent commercial living unrelated kidney transplantation suggests that although noncompliance is prevalent, it is not inevitable. Strategies to decrease noncompliance in young patients with chronic illnesses can be learned from the experience with transplant recipients. The general pediatrician has a central role in identifying and addressing the problem of noncompliance in adolescents with chronic disease. PMID- 15805373 TI - Primary hyperparathyroidism in pediatric patients. AB - OBJECTIVE: Primary hyperparathyroidism (HPT) is unusual in children. We reviewed our experience with HPT to better characterize these children. METHODS: The retrospective review of patients <19 years old who underwent parathyroid resection for primary HPT from 1970 to 2000 was performed at a single institution.. RESULTS: Fifty-two patients were identified. Median age was 16.8 years (range: 4-18.9) with a female-to-male ratio of 3:2. Eighty-five percent had an elevated parathyroid hormone (PTH) level, and 15% had an inappropriately normal PTH level during hypercalcemia. Serum calcium was elevated in all patients except for 2 with multiple endocrine neoplasma (MEN)-IIA and 1 with familial non MEN HPT, but both had elevated PTH levels. Alkaline phosphatase levels were significantly higher in children with documented bone involvement. At presentation 41 patients (79%) were symptomatic and end-organ damage (nephrocalcinosis, nephrolithiasis, acute pancreatitis, or bone involvement) occurred in 23 patients (44%). Thirty-four patients (65%) had a single adenoma; hyperplasia was identified in 16 patients (27%), and of these cases, 57% occurred in patients diagnosed with MEN-I. Short-term complications included transient hypocalcemia in 29 patients (56%) and transient vocal cord paralysis in 2 patients (4%). Long-term complications were significant for permanent hypocalcemia in 2 patients (4%) and no recurrent laryngeal nerve injuries. No parathyroid abnormalities were identified during exploration in 4 (8%) children. Long-term follow-up was achieved in 98% of patients for a mean and median of 13 years. Resolution of hypercalcemia was achieved in 94% of cases. CONCLUSION: The diagnosis of primary HPT in pediatric patients is frequently delayed, is commonly symptomatic, and has significant morbidity. For children in whom HPT is suspected, evaluation of serum calcium and PTH levels is diagnostic in 100% of children. Parathyroid resection is effective at restoring normal serum calcium, has few complications, and is the treatment of choice for children with primary hyperparathyroidism. PMID- 15805374 TI - Teen reach: outcomes from a randomized, controlled trial of a tobacco reduction program for teens seen in primary medical care. AB - OBJECTIVE: To test the long-term efficacy of brief counseling plus a computer based tobacco intervention for teens being seen for routine medical care. METHODS: Both smoking and nonsmoking teens, 14 to 17 years of age, who were being seen for routine visits were eligible for this 2-arm controlled trial. Staff members approached teens in waiting rooms of 7 large pediatric and family practice departments within a group-practice health maintenance organization. Of 3747 teens invited at > or =1 visits, 2526 (67%) consented and were randomized to tobacco intervention or brief dietary advice. The tobacco intervention was individually tailored on the basis of smoking status and stage of change. It included a 30-second clinician advice message, a 10-minute interactive computer program, a 5-minute motivational interview, and up to two 10-minute telephone or in-person booster sessions. The control intervention was a 5-minute motivational intervention to promote increased consumption of fruits and vegetables. Follow-up smoking status was assessed after 1 and 2 years. RESULTS: Abstinence rates after 2 years were significantly higher for the tobacco intervention arm, relative to the control group, in the combined sample of baseline smokers and nonsmokers (odds ratio [OR]: 1.23; 95% confidence interval [CI]: 1.03-1.47). Treatment effects were particularly strong among baseline self-described smokers (OR: 2.42; 95% CI: 1.40-4.16) but were not significant for baseline nonsmokers (OR: 1.25; 95% CI: 0.97-1.61) or for those who had "experimented" in the past month at baseline (OR: 0.95; 95% CI: 0.45-1.98). CONCLUSIONS: Brief, computer-assisted, tobacco intervention during routine medical care increased the smoking cessation rate among self-described smokers but was less effective in preventing smoking onset. PMID- 15805375 TI - Prenatal risk factors for severe retinopathy of prematurity among very preterm infants of the Australian and New Zealand Neonatal Network. AB - OBJECTIVE: To identify prenatal and perinatal risk factors for clinically severe (stage 3 or 4) retinopathy of prematurity (ROP). METHODS: Data were collected prospectively as part of the ongoing Australian and New Zealand Neonatal Network audit of high-risk infants (birth weight of <1500 g or gestational age [GA] of <32 weeks) admitted to a level III neonatal unit in Australia or New Zealand. Prenatal and perinatal factors to 1 minute of age were examined for the subset of infants with GA of <29 weeks who survived to 36 weeks' postmenstrual age and were examined for ROP (n = 2105). The factors significantly associated with stage 3 or 4 ROP were entered into a multivariate logistic regression model. RESULTS: Two hundred three infants (9.6%) had stage 3 or more ROP. Prematurity was the dominant risk factor, with infants with GA of <25 weeks having 20 times greater odds of severe ROP than infants with GA of 28 weeks. Birth weight for GA also had a "dose-response" effect; the more growth-restricted infants had greater risk, with infants below the 3rd percentile of weight for GA having 4 times greater odds of severe ROP than those between the 25th and 75th percentiles. Male gender was also a significant risk factor (odds ratio: 1.73; 95% confidence interval: 1.25-2.40). CONCLUSIONS: These data, for a large, essentially population-based cohort, suggest that factors related to the degree of immaturity, intrauterine growth restriction, and male gender contribute to severe ROP. PMID- 15805376 TI - Improved survival rates with increased neurodevelopmental disability for extremely low birth weight infants in the 1990s. AB - BACKGROUND: Advances in perinatal care have resulted in increased survival rates for extremely low birth weight children. We sought to examine the relative changes in rates of survival and neurodevelopmental impairment at 20 months of corrected age among 500- to 999-g birth weight infants born at our perinatal center during 2 periods, before and after the introduction of surfactant therapy in 1990. METHODS: Four hundred ninety-six infants with birth weights of 500 to 999 g were born at our perinatal center during period I (1982-1989) (mean body weight: 762 g; mean gestational age: 25.8 weeks) and 682 during period II (1990 1998) (mean body weight: 756 g; mean gestational age: 25.5 weeks). Rates of death and survival with and without neurodevelopmental impairment at 20 months of corrected age for the 2 periods were compared with logistic regression analyses, with adjustment for gestational age. RESULTS: Survival rates increased from 49% during period I to 67% during period II. Neonatal morbidity rates also increased during period II, including rates of sepsis (from 37% to 51%), periventricular leukomalacia (from 2% to 7%), and chronic lung disease, defined as oxygen dependence at 36 weeks of corrected age (from 32% to 43%). Rates of severe cranial ultrasound abnormalities were similar (22% vs 22%). Among children monitored, the rate of neurologic abnormalities, including cerebral palsy, increased from 16% during period I to 25% during period II and the rate of deafness increased from 3% to 7%. The overall rate of neurodevelopmental impairment (major neurosensory abnormality and/or Bayley Mental Developmental Index score of <70) increased from 26% to 36%. Compared with period I, in period II there were decreased rates of death (odds ratio [OR]: 0.3; 95% confidence interval [CI]: 0.2-0.4) and increased rates of survival with impairment (OR: 2.3; 95% CI: 1.7-3.3) but also increased rates of survival without impairment (OR: 1.7; 95% CI: 1.3-2.2). Compared with period I, for every 100 infants with birth weights of 500 to 999 g born in period II, 18 additional infants survived, of whom 7 were unimpaired and 11 were impaired. CONCLUSIONS: The improved survival rates in the 1990s occurred with an increased risk of significant neurodevelopmental impairment. Prospective parents of extremely low birth weight infants should be advised of this substantial risk, to facilitate decision-making in the delivery room. PMID- 15805377 TI - The role of endogenous opioids in mediating pain reduction by orally administered glucose among newborns. AB - OBJECTIVE: It has been demonstrated clearly that sweet-tasting solutions given before a painful intervention can reduce pain among newborns. There is no fully accepted explanation for this effect, but activation of endogenous opioids has been suggested as a possible mechanism. The aim of this study was to obtain deeper knowledge of the underlying mechanism by investigating whether administration of an opioid antagonist would reduce the effect of orally administered glucose at heel stick among term newborns. DESIGN: A randomized, placebo-controlled, double-blind trial with a validated, neonatal, pain-rating scale. PARTICIPANTS: The trial included 30 term newborns undergoing heel stick, who were assigned randomly to 1 of 2 groups, ie, group I, with naloxone hydrochloride (opioid antagonist) 0.01 mg/kg administered intravenously before oral administration of 1 mL of 30% glucose, or group II, with a corresponding amount of placebo (saline solution) administered intravenously before oral administration of glucose. OUTCOME MEASURES: Pain-related behavior during blood sampling was measured with the Premature Infants Pain Profile. Crying time and heart rate were also recorded. RESULTS: The 2 groups did not differ significantly in Premature Infant Pain Profile scores during heel stick. The median crying time during the first 3 minutes was 14 seconds (range: 0-174 seconds) for the naloxone group and 105 seconds (range: 0-175 seconds) for the placebo group. There was no significant difference in heart rate between the 2 groups. CONCLUSION: Administration of an opioid antagonist did not decrease the analgesic effect of orally administered glucose given before blood sampling. PMID- 15805378 TI - Early detection of imperforate hymen prevents morbidity from delays in diagnosis. AB - OBJECTIVE: Although it is detectable at all ages through inspection of the external genitalia, imperforate hymen (IH) is a diagnosis that is missed commonly. We hypothesized that children with late diagnoses (predefined as > or =8 years of age, chosen to reflect the timing of normal menarche) would be more likely to be symptomatic, undergo more diagnostic testing, and lack appropriate documentation in their medical records, compared with children with earlier diagnoses (ie, <8 years of age). METHODS: All patients with IH were identified through searches of 3 hospital databases with International Classification of Diseases, Ninth Revision, codes. The medical records of eligible subjects were reviewed by a single, blinded researcher. Comparisons were made between children diagnosed at younger versus older ages. RESULTS: A bimodal distribution of age at diagnosis was demonstrated; 43% (n = 10) of girls were diagnosed at <8 years of age, and 57% (n = 13) were diagnosed at > or =8 years of age. Among older girls, 100% were symptomatic (abdominal pain and/or urinary symptoms; duration of symptoms: 1-120 days), whereas 90% of cases in the younger group were detected incidentally. Documentation was lacking for breast development (77%), pubic hair development (69%), and menstrual history (46%) among the older girls. Older children were more likely to present symptomatically (odds ratio: 42.0; 95% confidence interval: 3.1-1965.7) and to undergo ancillary testing (odds ratio: 20.3; 95% confidence interval: 1.6-983.1). CONCLUSIONS: Two distinct populations of girls with IH exist, ie, those diagnosed without symptoms at a young age and those not diagnosed until >8 years of age. By incorporating an examination of the external genitalia into their routine practice, clinicians caring for children can prevent the significant delays in diagnosis, misdiagnosis, and morbidity associated with the latter group. PMID- 15805379 TI - Child health care clinicians' use of medications to help parents quit smoking: a national parent survey. AB - BACKGROUND: Smokers who use cessation medications when they attempt to quit double their likelihood of success. No prior survey has assessed the acceptability to parents of receiving smoking cessation medication prescriptions in the context of their child's primary care visits. OBJECTIVE: To assess acceptability to parents of receiving smoking cessation medication prescriptions and to compare that with the reported rate of actually receiving smoking cessation medication prescriptions in the context of the child's health care visit. METHODS: Data were collected through a national random-digit dial telephone survey of households from July to September 2003. The sample was weighted according to race and gender, on the basis of the 2002 US Census, to be representative of the US population. RESULTS: Of 3990 eligible respondents contacted, 3010 (75%) completed surveys; 1027 (34%) of those were parents. Of those parents, 211 (21%) were self-identified smokers. One half would consider using a smoking cessation medication and, of those, 85% said that it would be acceptable if the child's doctor prescribed or recommended it to them. In contrast, of the 143 smoking parents who accompanied their child to the doctor, only 15% had pharmacotherapy recommended and only 8% received a prescription for a smoking cessation medication. These results did not vary according to parent age, gender, race, or child age. CONCLUSIONS: Child health care clinicians have low rates of recommending and prescribing cessation therapies that have proved effective in other settings. The recommendation or provision of cessation medications would be acceptable to the majority of parents in the context of their child's health care visit. PMID- 15805380 TI - A multicenter, randomized, masked, comparison trial of lucinactant, colfosceril palmitate, and beractant for the prevention of respiratory distress syndrome among very preterm infants. AB - BACKGROUND AND OBJECTIVE: Evidence suggests that synthetic surfactants consisting solely of phospholipids can be improved through the addition of peptides, such as sinapultide, that mimic the action of human surfactant protein-B (SP-B). A synthetic surfactant containing a mimic of SP-B may also reduce the potential risks associated with the use of animal-derived products. Our objective was to compare the efficacy and safety of a novel synthetic surfactant containing a functional SP-B mimic (lucinactant; Discovery Laboratories, Doylestown, PA) with those of a non-protein-containing synthetic surfactant (colfosceril palmitate; GlaxoSmithKline, Brentford, United Kingdom) and a bovine-derived surfactant (beractant; Abbott Laboratories, Abbott Park, IL) in the prevention of neonatal respiratory distress syndrome (RDS) and RDS-related death. METHODS: We assigned randomly (double-masked) 1294 very preterm infants, weighing 600 to 1250 g and of < or =32 weeks gestational age, to receive colfosceril palmitate (n = 509), lucinactant (n = 527), or beractant (n = 258) within 20 to 30 minutes after birth. Primary outcome measures were the rates of RDS at 24 hours and the rates of death related to RDS during the first 14 days after birth. All-cause mortality rates, bronchopulmonary dysplasia (BPD) rates, and rates of other complications of prematurity were prespecified secondary outcomes. Primary outcomes, air leaks, and causes of death were assigned by an independent, masked, adjudication committee with prespecified definitions. The study was monitored by an independent data safety monitoring board. RESULTS: Lucinactant reduced significantly the incidence of RDS at 24 hours, compared with colfosceril (39.1% vs 47.2%; odds ratio [OR]: 0.68; 95% confidence interval [CI]: 0.52-0.89). There was no significant difference in comparison with beractant (33.3%). However, lucinactant reduced significantly RDS-related mortality rates by 14 days of life, compared with both colfosceril (4.7% vs 9.4%; OR: 0.43; 95% CI: 0.25-0.73) and beractant (10.5%; OR: 0.35; 95% CI: 0.18-0.66). In addition, BPD at 36 weeks postmenstrual age was significantly less common with lucinactant than with colfosceril (40.2% vs 45.0%; OR: 0.75; 95% CI: 0.56-0.99), and the all-cause mortality rate at 36 weeks postmenstrual age was lower with lucinactant than with beractant (21% vs 26%; OR: 0.67; 95% CI: 0.45-1.00). CONCLUSIONS: Lucinactant is a more effective surfactant preparation than colfosceril palmitate for the prevention of RDS. In addition, lucinactant reduces the incidence of BPD, compared with colfosceril palmitate, and decreases RDS-related mortality rates, compared with beractant. Therefore, we conclude that lucinactant, the first of a new class of surfactants containing a functional protein analog of SP-B, is an effective therapeutic option for preterm infants at risk for RDS. PMID- 15805381 TI - A multicenter, randomized, controlled trial of lucinactant versus poractant alfa among very premature infants at high risk for respiratory distress syndrome. AB - BACKGROUND: Available therapeutic surfactants are either animal-derived or non protein-containing synthetic products. Animal-derived surfactants contain variable amounts of surfactant apoproteins, whereas the older-generation synthetic products contain only phospholipids and lack surfactant proteins (SPs). Both decrease morbidity and mortality rates associated with respiratory distress syndrome (RDS) among preterm infants, compared with placebo. However, excess mortality rates have been observed with non-protein-containing synthetic surfactants, compared with the animal-derived products. Evidence suggests that synthetic surfactants consisting solely of phospholipids can be improved with the addition of peptides that are functional analogs of SPs. Lucinactant is a new synthetic peptide-containing surfactant that contains sinapultide, a novel, 21 amino acid peptide (leucine and lysine repeating units, KL4 peptide) designed to mimic human SP-B. It is completely devoid of animal-derived components. OBJECTIVE: We hypothesized that the outcomes for premature infants treated with lucinactant and poractant alfa would be similar. Therefore, we compared lucinactant (Surfaxin; Discovery Laboratories, Doylestown, PA) with porcine derived, poractant alfa (Curosurf; Chiesi Farmaceutici, Parma, Italy) in a trial to test for noninferiority. METHODS: A total of 252 infants born between 24 and 28 weeks of completed gestation, with birth weights between 600 and 1250 g, were assigned randomly in a multicenter, multinational, noninferiority, randomized, controlled study to receive either lucinactant (n = 124) or poractant alfa (n = 128) within 30 minutes of life. The primary outcome was the incidence of being alive without bronchopulmonary dysplasia (BPD) through 28 days of age. Key secondary outcomes included death at day 28 and 36 weeks postmenstrual age (PMA), air leaks, neuroimaging abnormalities, and other complications related to either prematurity or RDS. An independent, international, data and safety monitoring committee monitored the trial. RESULTS: The treatment difference between lucinactant and poractant alfa for survival without BPD through 28 days was 4.75% (95% confidence interval [CI]: -7.3% to 16.8%) in favor of lucinactant, with the lower boundary of the 95% CI for the difference, ie, -7.3%, being greater than the prespecified noninferiority margin of -14.5%. At 28 days, 45 of 119 infants given lucinactant were alive without BPD (37.8%; 95% CI: 29.1-46.5%), compared with 41 of 124 given poractant alfa (33.1%; 95% CI: 24.8-41.3%); at 36 weeks PMA, the rates were 64.7% and 66.9%, respectively. The corresponding mortality rate through day 28 for the lucinactant group was lower than that for the poractant alfa group (11.8% [95% CI: 6.0-17.6%] vs 16.1% [95% CI: 9.7-22.6%]), as was the rate at 36 weeks PMA (16% and 18.5%, respectively). There were no differences in major dosing complications. In addition, no significant differences were observed in the incidences of common complications of prematurity, including intraventricular hemorrhage (grades 3 and 4) and cystic periventricular leukomalacia (lucinactant: 14.3%; poractant alfa: 16.9%). CONCLUSIONS: Lucinactant and poractant alfa were similar in terms of efficacy and safety when used for the prevention and treatment of RDS among preterm infants. The ability to enhance the performance of a synthetic surfactant with the addition of a peptide that mimics the action of SP-B, such as sinapultide, brings potential advantages to exogenous surfactant therapy. PMID- 15805382 TI - A comparison of 2 influenza vaccine schedules in 6- to 23-month-old children. AB - BACKGROUND: Trivalent inactivated influenza vaccine (TIV) is recommended for all children ages 6 to 23 months. Delivering 2 doses of TIV at least 4 weeks apart to young children receiving this vaccine for the first time is challenging. METHODS: We compared the immunogenicity and reactogenicity of the standard 2-dose regimen of TIV administered in the fall with an early schedule of a single spring dose followed by a fall dose of the same vaccine in healthy toddlers 6 to 23 months of age. Children were recruited in the spring to be randomized into either the standard or early schedule. An additional group was also enrolled in the fall as part of a nonrandomized standard comparison group. The 2002-2003 licensed TIV was administered in the spring; the fall 2003-2004 vaccine contained the same 3 antigenic components. Reactogenicity was assessed by parental diaries and telephone surveillance. Blood was obtained after the second dose of TIV for all children. The primary outcome measure was antibody response to influenza A/H1N1, A/H3N2, and B after 2 doses of vaccine, as determined by hemagglutination inhibition titers > or =1:32 and geometric mean titer (GMT). RESULTS: Two hundred nineteen children were randomized to receive either the standard or early TIV schedule; 40 additional children were enrolled in the fall in the nonrandomized standard group. Response rates in the combined standard versus early groups were similar overall: 78% (GMT: 48) vs 76% (GMT: 57) to H1N1, 89% (GMT: 115) vs 88% (GMT: 129) to H3N2, and 52% (GMT: 24) vs 60% (GMT: 28) to B. Reactogenicity after TIV in both groups of children was minimal and did not differ by dose, age, or time between doses. Reaction rates were higher in those receiving TIV and concomitant vaccines compared with those receiving TIV alone. Overall rates of fever >38 degrees C axillary and injection-site pain, redness, or swelling were 5.4%, 3.1%, 0.9%, and 1.1%, respectively. CONCLUSIONS: When the spring and fall influenza vaccines had the same 3 antigenic components, the early vaccine schedule resulted in similar immunogenicity and reactogenicity compared with the standard schedule. When the vaccine components do not change between years, initiating influenza vaccine in the spring at the time of routine office visits would facilitate full immunization of children against influenza earlier in the season. PMID- 15805383 TI - A review of evidence supporting the American Academy of Pediatrics recommendation for prescribing cephalosporin antibiotics for penicillin-allergic patients. AB - The American Academy of Pediatrics, evidence-based guidelines endorse the use of cephalosporin antibiotics for patients with reported allergies to penicillin, for the treatment of acute bacterial sinusitis and acute otitis media. Many physicians, however, remain reluctant to prescribe such agents. Although such concern is understandable, lack of consistent data regarding exactly what constitutes an initial penicillin-allergic reaction and subsequent cross sensitivity to cephalosporins may be preventing many patients from receiving optimal antibiotic therapy. This article reviews evidence in support of the American Academy of Pediatrics recommendation. Included is an examination of the types and incidence of reactions to penicillins and cephalosporins; the frequency of cross-reactivity between these 2 groups of agents; experimental and clinical studies that suggest that side chain-specific antibodies predominate in the immune response to cephalosporins, thereby explaining the lack of cross sensitivity between most cephalosporins and penicillins; the role of skin testing; and the risks of anaphylaxis. Specific recommendations for the treatment of patients on the basis of their responses to previously prescribed agents are summarized. PMID- 15805384 TI - Challenge of transforming our private and public pediatric health care systems to emphasize value. PMID- 15805385 TI - Heart rate characteristics in neonatal sepsis: a promising test that is still premature. PMID- 15805386 TI - Neonatal resuscitation: what role for volume expansion? PMID- 15805387 TI - Procedural pain in neonates: the new millennium. PMID- 15805388 TI - Primary hyperparathyroidism in pediatric patients: clear-cut differences from adult patients. PMID- 15805389 TI - Synthetic surfactants: the search goes on. PMID- 15805390 TI - Delinquent youth in corrections: Medicaid and reentry into the community. PMID- 15805391 TI - Structure versus function: time will always tell. PMID- 15805392 TI - Kids and motorbikes: the need for speed. PMID- 15805393 TI - Motorized vehicles for children: a new public health problem. PMID- 15805394 TI - To treat or not to treat. PMID- 15805395 TI - Twenty years of emergency medical services for children: a cause for celebration and a call for action. PMID- 15805396 TI - The pediatrician's role in community pediatrics. AB - This policy statement reaffirms the pediatrician's role in community pediatrics. It offers pediatricians a definition of community pediatrics and provides a set of specific recommendations that underscore the critical nature of this important dimension of the profession. PMID- 15805397 TI - Providing care for immigrant, homeless, and migrant children. AB - This policy statement, which replaces the retired statements "Health Care for Children of Immigrant Families" (1997) and "Health Needs of Homeless Children and Families" (1996), is a broader discussion and addresses not only immigrant but also homeless and migrant child populations. It provides pediatricians with the necessary framework for addressing underserved children: those who face substantial barriers that limit access to appropriate health care services. This statement supports a community-based approach to health care delivery to ensure that underserved children have a medical home. PMID- 15805398 TI - Guiding principles for pediatric hospitalist programs. AB - Pediatric hospitalist programs have become increasingly popular recently, emulating the growth and success of adult hospitalist programs. This statement provides an overview of hospitalist programs, factors influencing their growth, and their expected benefits. Six guiding principles for the establishment of pediatric hospitalist programs are identified in this statement concerning voluntary referrals; local design; minimum physician-training requirements; arrangement for appropriate follow-up; communication among primary care physicians, subspecialists, and hospitalists; and data collection and outcome measurements. PMID- 15805399 TI - Use of performance-enhancing substances. AB - Performance-enhancing substances include dietary supplements, prescription medications, and illicit drugs. Virtually no data are available on the efficacy and safety in children and adolescents of widely used performance-enhancing substances. This statement is intended to provide a generalized but functional definition of performance-enhancing substances. The American Academy of Pediatrics strongly condemns the use of performance-enhancing substances and vigorously endorses efforts to eliminate their use among children and adolescents. PMID- 15805401 TI - Use of serum electrolyte panels in gastroenteritis. PMID- 15805402 TI - Use of serum electrolyte panels in gastroenteritis. PMID- 15805404 TI - Use of serum electrolyte panels in gastroenteritis. PMID- 15805405 TI - The natural course of infantile reflux regurgitation: a non-Western perspective. PMID- 15805406 TI - Brain-type natriuretic peptide in the diagnosis and management of persistent pulmonary hypertension of the newborn. PMID- 15805408 TI - Detection of the Bartonella henselae gene sequence in lymph nodes of children with Kikuchi's disease. PMID- 15805411 TI - Can the results be believed? PMID- 15805412 TI - Should officers of the US Department of Health and Human Services advocate pet theories of sudden infant death syndrome? PMID- 15805413 TI - Preterm circulatory support is more complex than just blood pressure. PMID- 15805416 TI - Stones from a glass house. PMID- 15805418 TI - Multiple roles for elastic fibers in the skin. AB - Dermal elastic fibers are believed to have a primary role in providing elastic stretch and recoil to the skin. Here we compare the structural arrangement of dermal elastic fibers of chick skin and different animal species. Most elastic fibers in chick skin are derived from cells that line the feather follicle and/or smooth muscle that connects the pterial and apterial muscle bundles to feather follicles. Elastic fibers in the dermis of animals with single, primary hair follicles are derived from cells lining the hair follicle or from the ends of the pili muscle, which anchors the muscle to the matrix or to the hair follicle. Each follicle is interconnected with elastic fibers. Follicles of animals with primary and secondary (wool) hair follicles are also interconnected by elastic fibers, yet only the elastic fibers derived from the primary follicle are connected to each primary follicle. Only the primary hair follicles are connected to the pili muscle. Human skin, but not the skin of other primates, is significantly different from other animals with respect to elastic fiber organization and probably cell of origin. The data suggest that the primary role for elastic fibers in animals, with the possible exception of humans, is movement and/or placement of feathers or hair. PMID- 15805417 TI - A novel marker for Purkinje cells, KIAA0864 protein. An analysis based on a monoclonal antibody HFB-16 in developing human cerebellum. AB - In the search for immunohistochemical markers of the developing human brain, a monoclonal antibody, HFB-16, was raised against homogenates from the cerebrum of a 15-gestational-week-old (GW) human fetus and screened on paraffin-embedded human embryonic brain specimens. This antibody was particularly useful as a marker for Purkinje cells in the developing human cerebellum. Positive immunoreactivities with HFB-16 first appeared in the Purkinje cell layer at 17 GW. From 20 to 24 GW, positive immunoreactivities were found above the lamina dissecans. After 25 GW, dendrites of Purkinje cells were found with the HFB-16 antibody, and the nerve fibers of the Purkinje cells became positive after 35 GW. Neurons in the dentate nucleus and external and internal granular layers reacted negatively to this antibody. After 1 year, when the external granular layer faded out, the dendrites of the Purkinje cells reached the pial surface of the cerebellum, and nerve fibers began to develop in the white matter. This antibody was also useful for characterization of components in heterotopic neurons found in various anomaly syndromes such as trisomy 13. Expressional cloning indicated the antigen against HFB-16 to be human KIAA0864 protein, which is supposed to be an alternative splicing product of p116Rip, whose function has not yet been elucidated. The antigenicity of the KIAA0864 protein was confirmed using human cDNA of the KIAA0864 protein, a protein expression vector, and an HFB-16 antibody. PMID- 15805419 TI - Fiber content and myosin heavy chain composition of muscle spindles in aged human biceps brachii. AB - The present study investigated potential age-related changes in human muscle spindles with respect to the intrafusal fiber-type content and myosin heavy chain (MyHC) composition in biceps brachii muscle. The total number of intrafusal fibers per spindle decreased significantly with aging, due to a significant reduction in the number of nuclear chain fibers. Nuclear chain fibers in old spindles were short and some showed novel expression of MyHC alpha-cardiac. The expression of MyHC alpha-cardiac in bag1 and bag2 fibers was greatly decreased in the A region. The expression of slow MyHC was increased in nuclear bag1 fibers and that of fetal MyHC decreased in bag2 fibers whereas the patterns of distribution of the remaining MyHC isoforms were generally not affected by aging. We conclude that aging appears to have an important impact on muscle spindle composition. These changes in muscle spindle phenotype may reflect an age-related deterioration in sensory and motor innervation and are likely to have an impact in motor control in the elderly. PMID- 15805420 TI - Chromatoid bodies: aggresome-like characteristics and degradation sites for organelles of spermiogenic cells. AB - We investigated the localization of several markers for lysosomes and aggresomes in the chromatoid bodies (CBs) by immunoelectron microscopy. We found so-called aggresomal markers such as Hsp70 and ubiquitin in the core of the CBs and vimentin and proteasome subunit around the CBs. Ubiquitin-conjugating enzyme (E2) was also found in the CBs. In tubulovesicular structures surrounding the CBs, lysosomal markers were detected but an endoplasmic reticulum retention signal (KDEL) was not. Moreover, proteins located in each subcellular compartment, including the cytosol, mitochondria, and nucleus, were detected in the CBs. Signals for cytochrome oxidase I (COXI) coded on mitochondrial DNA were also found in the CBs. Quantitative analysis of labeling density showed that all proteins examined were concentrated in the CBs to some extent. These results show that the CBs have some aggresomal features, suggesting that they are not a synthetic site as proposed previously but a degradation site where unnecessary DNA, RNA, and proteins are digested. PMID- 15805421 TI - Immunocytochemistry for drugs containing an aliphatic primary amino group in the molecule, anticancer antibiotic daunomycin as a model. AB - An immunocytochemistry (ICC) for the anticancer antibiotic daunomycin (DM) was developed using a combination of anti-DM serum produced against N-(gamma maleimidobutyryloxy)succinimide (GMBS)-conjugated DM, and DM-uptake human melanoma BD cells. The antiserum was demonstrated to be specific for DM and the structurally related analogs adriamicin and epirubicin by an ICC model system of the enzyme immunoassay (EIA) using glutaraldehyde (GA)-conjugated DM as a solid phase antigen. No cross-reaction occurred with any of the other antibiotics tested such as bleomycin, pepleomycin, and mitomycin C. Successful DM ICC required a series of processes prior to the immunocytochemical reaction: the cells were first fixed with GA, then reduced with NaBH4, treated with hydrochloric acid, and finally digested with protease. The cell specimens were then subjected to immunoreaction with anti-DM serum followed by peroxidase labeled goat anti-rabbit IgG/Fab', and in both immune reagents the detergent Triton X-100 was contained as well. The present ICC covering all these processes successfully stained for DM in the nucleus and in the perinuclear Golgi region of the cytoplasm of the BD cells, consistent with the results obtained by the DM autofluorescence method. This ICC was found to be three times as sensitive as the cytofluorometric method and applicable to the paraffin sections of the liver of rats 24 hr after an IV injection of DM. The principle used in the present study for developing DM ICC might be applied to other drugs containing the primary amino group(s) in the molecule. Thus, these ICCs for drugs are direct, precise and easy new methods that should have potential for pharmacology and toxicology studies of drugs, revealing the localization of a drug in cells and tissues. PMID- 15805422 TI - The LIM domain protein Lmo4 is highly expressed in proliferating mouse epithelial tissues. AB - LMO4 belongs to the LIM-only family of zinc finger proteins that have been implicated in oncogenesis. The LMO4 gene is overexpressed in breast cancer and oral cavity carcinomas, and high levels of this protein inhibit mammary epithelial differentiation. Targeted deletion of Lmo4 in mice leads to complex phenotypic abnormalities and perinatal lethality. To further understand the role of LMO4, we have characterized Lmo4 expression in adult mouse tissues by immunohistochemical staining using monoclonal anti-Lmo4 antibodies. Lmo4 was highly expressed within specific cell types in diverse tissues. Expression was prevalent in epithelial-derived tissues, including the mammary gland, tongue, skin, small intestine, lung, and brain. High levels of Lmo4 were frequently observed in proliferating cells, such as the crypt cells of the small intestine and the basal cells of the skin and tongue. Lmo4 was highly expressed in the proliferative cap cell layer of the terminal end buds in the peripubertal mammary gland and in the lobuloalveolar units during pregnancy. The expression profile of Lmo4 suggests that this cofactor is an important regulator of epithelial proliferation and has implications for its role in the pathogenicity of cancer. PMID- 15805423 TI - Altered expression patterns of heterogeneous nuclear ribonucleoproteins A2 and B1 in the adrenal cortex. AB - Several proteins implicated in hormonogenesis of the adrenal cortex have alternatively spliced isoforms, which respond differently to adrenocorticotropic hormone (ACTH). Heterogeneous nuclear ribonucleoproteins A2 and B1 are among the abundant pre-mRNA-binding proteins involved in alternative splicing. We examined the expression of A2 and B1 in normal adrenal cortex and tumors. B1 was variably expressed in the zona fasciculata-reticularis, although A2 was diffusely expressed in the three zones. B1 was more abundant in compact cells than clear cells, and B1 expression was frequent in the zona reticularis, which consists mainly of compact cells. In three kinds of cortical adenomas autonomously producing hormones, B1 was generally overexpressed and there were no significant differences among them. In cortisol-producing tumors, non-tumor parts of the cortex, which were generally atrophic due to low ACTH, had less B1 protein than normal adrenals. These results suggested a correlation between B1 expression and the hormonal activity responding to ACTH. In vitro ACTH stimulation induced a biphasic expression of B1 in an H295R cortical carcinoma cell line, and it paralleled hormonogenesis. Conclusively, B1 expression varied in relation to the hormonal activity responding to the ACTH, and it may provide a key to elucidating the splicing mechanisms involved in hormonogenesis. PMID- 15805424 TI - Visualization of the nuclear lamina in mouse anterior pituitary cells and immunocytochemical detection of lamin A/C by quick-freeze freeze-substitution electron microscopy. AB - We examined the nuclear lamina in the quickly frozen anterior pituitary cells by electron microscopic techniques combined with freeze substitution, deep etching, and immunocytochemistry and compared it with that in the chemically fixed cells. By quick-freeze freeze-substitution electron microscopy, an electron-lucent layer, as thick as 20 nm, was revealed just inside the inner nuclear membrane, whereas in the conventionally glutaraldehyde-fixed cells the layer was not seen. By quick-freeze deep-etch electron microscopy, we could not distinguish definitively the layer corresponding to the nuclear lamina in either fresh unfixed or glutaraldehyde-fixed cells. Immunofluorescence microscopy showed that lamin A/C in the nucleus was detected in the acetone-fixed cells and briefly in paraformaldehyde-fixed cells but not in the cells with prolonged paraformaldehyde fixation. Nuclear localization of lamin A/C was revealed by immunogold electron microscopy also in the quickly frozen and freeze-substituted cells, but not in the paraformaldehyde-fixed cells. Lamin A/C was localized mainly in the peripheral nucleoplasm within 60 nm from the inner nuclear membrane, which corresponded to the nuclear lamina. These results suggest that the nuclear lamina can be preserved both ultrastructurally and immunocytochemically by quick freezing fixation, rather than by conventional chemical fixation. PMID- 15805425 TI - Immunohistochemical expression of p16INK4a and bcl-2 according to HPV type and to the progression of cervical squamous intraepithelial lesions. AB - Inactivation of the cell cycle inhibitor gene p16MTS1 seems to be involved in human papillomavirus (HPV)-related carcinogenesis because E6 and E7 oncoproteins may impair p16INK4a and, indirectly, bcl-2 functions. In this study, we analyzed the role of immunohistochemical expression of p16INK4a and bcl-2 in HPV-infected cervical biopsies as prognostic markers of the progression of squamous intraepithelial lesion (SIL). Sixty-five cervical biopsies were stratified into two subgroups according to the second biopsy: 27 of them maintained a low-grade (LG)-SIL diagnosis, and 38 progressed from LG-SIL to high-grade (HG)-SIL. p16INK4a and bcl-2 quantitative expression levels were measured by the immunoperoxidase method. PCR-DNA techniques were used to detect and type HPV. The Wilcoxon and Fisher exact tests were employed for the statistical analysis. In the group with an LG-SIL diagnosis at the second biopsy, no significant associations were found between p16INK4a and bcl-2 expression and presence of HPV16/18. In the group that progressed to HG-SIL, a significant association was observed between p16INK4a overexpression and HPV16/18 presence (p=0.021), but none with bcl-2 levels. It is concluded that immunohistochemical bcl-2 expression may not be useful for predicting the progression of HPV-related SIL. In contrast, p16INK4a overexpression seemed to be associated with HPV 16 and 18, suggesting that it may be a good marker for predicting SIL progression. PMID- 15805426 TI - Immunohistochemical localization of P-glycoprotein in rat brain and detection of its increased expression by seizures are sensitive to fixation and staining variables. AB - The MDR1 gene product, P-glycoprotein (P-gp), was shown to confer multidrug resistance to cancer cells, but its overexpression is also suggested to be involved in pharmacoresistance of epilepsy by acting as an energy-dependent drug efflux pump in the blood-brain barrier (BBB). In normal brain tissue, P-gp is almost exclusively expressed by capillary endothelial cells (EC) of the BBB, whereas little or no expression is detected in other cell types. Increased P-gp expression was observed after seizures, but localization of this increase, i.e., within brain capillary EC or within parenchymal or perivascular astrocytes, which contribute to the BBB function, is controversial. To test whether these antithetic data arise from unusual properties of the antigen itself, we compared different immunohistochemical techniques and monoclonal or polyclonal antibodies to P-gp in normal rat brain and rat brain after kainate-induced seizures. Using acetone-fixed cryostat sections of snap-frozen tissue, strong P-gp labeling was detected in EC and, after seizures, in hippocampal neurons, but not in astrocytes. In contrast, EC and neuronal P-gp immunolabeling were not seen in paraformaldehyde-fixed sections, whereas both perivascular and parenchymal astrocytes exhibited strong P-gp labeling after seizures. The lack of P-gp labeling in EC by paraformaldehyde fixation, was reversed by treatment of the sections with acetate/ethanol. These experiments demonstrate that various fixation conditions have a striking effect on the immunohistochemical localization of P-gp in rat brain and detection of its increased expression by seizures. When data obtained from different immunohistochemical techniques are taken together, seizures seem to induce overexpression of P-gp in four different cell types, i.e., EC, perivascular astrocytes, parenchymal astrocytes, and neurons. PMID- 15805427 TI - Mitochondrial expression of arginase II in male and female rat inner medullary collecting ducts. AB - Microdissected rat proximal straight tubules (PST) and inner medullary collecting ducts (IMCD) highly produce urea from l-arginine, supporting the expression of the mitochondrial arginase II. However, IMCD contain a very low density of mitochondria compared with PST. Recently, arginase II has been localized by immunohistochemistry in rat PST but not IMCD. This study was designed to verify whether rat IMCD express arginase II and to identify its subcellular localization. We developed an antibody raised against arginase II that allowed the detection of a band of 38 kDa corresponding to arginase II on immunoblots. In male and female rat kidneys, Western blot analyses revealed that arginase II was highly expressed in the inner medulla (IM), the outer stripe of the outer medulla (osOM), and the deep cortex. Immunocytochemistry demonstrated that arginase II was homogeneously expressed in IMCD. Proteins of the cytosolic and mitochondrial fractions extracted from osOM and IM and analyzed by Western blot showed that 86% of arginase II was associated with mitochondria. The molecular weight of arginase II was similar in the cytosolic and mitochondrial fractions. Immunoelectron microscopy confirmed the presence of arginase II in the mitochondria of IMCD. In conclusion, arginase II is expressed in mitochondria of male and female rat IMCD. PMID- 15805428 TI - Monoclonal antibodies to plant cell wall xylans and arabinoxylans. AB - Two rat monoclonal antibodies have been generated to plant cell wall (1-->4)-beta D-xylans using a penta-1,4-xylanoside-containing neoglycoprotein as an immunogen. The monoclonal antibodies, designated LM10 and LM11, have different specificities to xylans in relation to the substitution of the xylan backbone as indicated by immunodot assays and competitive-inhibition ELISAs. LM10 is specific to unsubstituted or low-substituted xylans, whereas LM11 binds to wheat arabinoxylan in addition to unsubstituted xylans. Immunocytochemical analyses indicated the presence of both epitopes in secondary cell walls of xylem but differences in occurrence in other cell types. PMID- 15805431 TI - Towards an evidence based prevention of sports injuries. PMID- 15805429 TI - The origins of diversity and specificity in g protein-coupled receptor signaling. AB - The modulation of transmembrane signaling by G protein-coupled receptors (GPCRs) constitutes the single most important therapeutic target in medicine. Drugs acting on GPCRs have traditionally been classified as agonists, partial agonists, or antagonists based on a two-state model of receptor function embodied in the ternary complex model. Over the past decade, however, many lines of investigation have shown that GPCR signaling exhibits greater diversity and "texture" than previously appreciated. Signal diversity arises from numerous factors, among which are the ability of receptors to adopt multiple "active" states with different effector-coupling profiles; the formation of receptor dimers that exhibit unique pharmacology, signaling, and trafficking; the dissociation of receptor "activation" from desensitization and internalization; and the discovery that non-G protein effectors mediate some aspects of GPCR signaling. At the same time, clustering of GPCRs with their downstream effectors in membrane microdomains and interactions between receptors and a plethora of multidomain scaffolding proteins and accessory/chaperone molecules confer signal preorganization, efficiency, and specificity. In this context, the concept of agonist-selective trafficking of receptor signaling, which recognizes that a bound ligand may select between a menu of active receptor conformations and induce only a subset of the possible response profile, presents the opportunity to develop drugs that change the quality as well as the quantity of efficacy. As a more comprehensive understanding of the complexity of GPCR signaling is developed, the rational design of ligands possessing increased specific efficacy and attenuated side effects may become the standard mode of drug development. PMID- 15805432 TI - Binge drinking. PMID- 15805430 TI - Cancer prevention: a new era beyond cyclooxygenase-2. AB - The seminal epidemiological observation that nonsteroidal anti-inflammatory drugs (NSAIDs) prevent colon and possibly other cancers has spurred novel approaches to cancer prevention. The known inhibitory effect of NSAIDs on the eicosanoid pathway prompted studies focusing on cyclooxygenase (COX) and its products. The increased prostaglandin E2 levels and the overexpression of COX-2 in colon and many other cancers provided the rationale for clinical trials with COX-2 inhibitors for cancer prevention or treatment. Their efficacy in the prevention of sporadic colon and other cancers remains unknown; one COX-2 inhibitor has been withdrawn because of side effects, and there are concerns about whether these effects are class-specific. There is evidence to suggest that COX-2 may not be the only or ideal eicosanoid pathway target for cancer prevention. Six sets of observations support this notion: the relatively late induction of COX-2 during carcinogenesis; the finding that NSAIDs may not require inhibition of COX-2 for their effect; the modest effect of coxibs in cancer prevention; that currently available coxibs have multiple non-COX-2 effects that may account for at least some of their efficacy; the possibility that concurrent inhibition of COX-2 in non-neoplastic cells may be harmful; and the possibility that COX-2 inhibition may modulate alternative eicosanoid pathways in a way that promotes carcinogenesis. Given the limitations of COX-2-specific inhibitors and the biological evidence mentioned above, we suggest that targets other than COX-2 should be pursued as alternative or complementary approaches to cancer prevention. PMID- 15805433 TI - Winning, losing, and violence. AB - BACKGROUND: Although international sports events attract huge interest, and results can be a barometer of popular national standing, their impact on violent behaviour has not been investigated. METHODS: Associations between assault related emergency department (ED) attendances and international sporting events (home and away rugby and soccer matches) in a European capital city (Cardiff) served by one ED, between 1 May 1995 and 30 April 2002 were investigated. The frequency of assault related ED attendances were studied relative to whether the national team won or lost, controlling for potential covariates: match attendance, match location (home/away), results (win/lose), net scores, and day of match (weekend/weekday). Multiple linear regression was used to identify significant associations with ED assault related attendances. RESULTS: Matches which the Wales team won (p = 0.03), match attendance (p<0.001), and weekend matches (p<0.001) were positively associated with ED assault related injury attendances. Assault frequency measured in this way was no different for home and away matches. CONCLUSIONS: Assault injury resulting in ED treatment was more frequent when national teams won than when they lost. Sport type made no difference. Violence prevention efforts should be increased on international match days, when the national team is expected to win, when match attendance is large, and for away as well as home matches. PMID- 15805434 TI - Comparison of injury case fatality rates in the United States and New Zealand. AB - OBJECTIVE: To compare injury case fatality rates in the United States (US) with New Zealand (NZ) to guide future information collection, research, and evaluation. DESIGN: Using NZ (1992-96) and US (1996-98) mortality censuses, NZ national 1992-96 hospital discharge censuses, and US 1996-98 National Hospital Discharge Survey data, the authors compared case fatality rates by mechanism and intent of injury and age group. The analysis was restricted to severe injuries (AIS> or =3). SUBJECTS: NZ (1992-96) and US (1996-98) populations. MAIN OUTCOME MEASURES: Ratio of case fatality rates in NZ versus the US (RCFR(NZ:US)). RESULTS: Overall, among cases meeting the study criteria, unintentional injuries were 1.57 times more likely fatal in NZ and intentional assault injuries were 1.14 times more likely to be fatal in the US. Firearms were involved in 50% of US assaults versus 8% of NZ assaults. By mechanism, cutting/piercing injuries were 1.86, firearm injuries were 1.41, and motor vehicle injuries were 1.44 times more to be likely fatal in NZ. Natural/environmental injuries (RCFR(NZ:US) = 0.57), unintentional poisonings (RCFR(NZ:US) = 0.26), and unintentional suffocations (RCFR(NZ:US) = 0.67) were significantly more likely to be fatal in the US. CONCLUSIONS: Possible reasons for the observed results include: differences in geography and proportion of population in rural areas, trauma system differences, road design and vehicle types, seat belt use, larger role of firearms in US assaults, coding practices, policies, and environmental factors. Disparities evoke hypotheses to test in future research that will guide priority setting and intervention. PMID- 15805435 TI - An evaluation of state firearm regulations and homicide and suicide death rates. AB - OBJECTIVE: To determine if any of five different state gun laws were associated with firearm mortality: (1) "shall issue" laws permitting an individual to carry a concealed weapon unless restricted by another statute; (2) a minimum age of 21 years for handgun purchase; (3) a minimum age of 21 years for private handgun possession; (4) one gun a month laws which restrict handgun purchase frequency; and (5) junk gun laws which ban the sale of certain cheaply constructed handguns. DESIGN: A cross sectional time series study of firearm mortality from 1979 to 1998. SETTING: All 50 states and the District of Columbia. SUBJECTS: All residents of the United States. MAIN OUTCOME MEASURES: Firearm homicides, all homicides, firearm suicides, and all suicides. RESULTS: When a "shall issue" law was present, the rate of firearm homicides was greater, RR 1.11 (95% confidence interval 0.99 to 1.24), than when the law was not present, as was the rate of all homicides, RR 1.08 (95% CI 0.98 to 1.17), although this was not statistically significant. No law was associated with a statistically significant decrease in the rates of firearm homicides or total homicides. No law was associated with a statistically significant change in firearm suicide rates. CONCLUSION: A "shall issue" law that eliminates most restrictions on carrying a concealed weapon may be associated with increased firearm homicide rates. No law was associated with a statistically significant reduction in firearm homicide or suicide rates. PMID- 15805436 TI - Effects on alcohol related fatal crashes of a community based initiative to increase substance abuse treatment and reduce alcohol availability. AB - OBJECTIVE: This analysis tested whether comprehensive community interventions that focus on reducing alcohol availability and increasing substance abuse treatment can reduce alcohol related fatal traffic crashes. INTERVENTION: Five of 14 communities awarded Fighting Back grants by The Robert Wood Johnson Foundation to reduce substance abuse and related problems attempted to reduce availability of alcohol and expand substance abuse treatment programs (FBAT communities). Program implementation began on 1 January 1992. DESIGN: A quasi-experimental design matched each program community to two or three other communities of similar demographic composition in the same state. MAIN OUTCOME MEASURES: The ratio of fatal crashes involving a driver or pedestrian with a blood alcohol concentration of 0.01% or higher, 0.08% or higher, or 0.15% or higher were examined relative to fatal crashes where no alcohol was involved for 10 years preceding and 10 years following program initiation. RESULTS: Relative to their comparison communities, the five FBAT communities experienced significant declines of 22% in alcohol related fatal crashes at 0.01% BAC or higher, 20% at 0.08% or higher, and 17% at 0.15% or higher relative to fatal crashes not involving alcohol. CONCLUSIONS: Community interventions to reduce alcohol availability and increase substance abuse treatment can reduce alcohol related fatal traffic crashes. PMID- 15805437 TI - Incidence of sports and recreation related injuries resulting in hospitalization in Wisconsin in 2000. AB - OBJECTIVE: To describe the incidence and patterns of sports and recreation related injuries resulting in inpatient hospitalization in Wisconsin. Although much sports and recreation related injury research has focused on the emergency department setting, little is known about the scope or characteristics of more severe sports injuries resulting in hospitalization. SETTING: The Wisconsin Bureau of Health Information (BHI) maintains hospital inpatient discharge data through a statewide mandatory reporting system. The database contains demographic and health information on all patients hospitalized in acute care non-federal hospitals in Wisconsin. METHODS: The authors developed a classification scheme based on the International Classification of Diseases External cause of injury code (E code) to identify hospitalizations for sports and recreation related injuries from the BHI data files (2000). Due to the uncertainty within E codes in specifying sports and recreation related injuries, the authors used Bayesian analysis to model the incidence of these types of injuries. RESULTS: There were 1714 (95% credible interval 1499 to 2022) sports and recreation-related injury hospitalizations in Wisconsin in 2000 (32.0 per 100,000 population). The most common mechanisms of injury were being struck by/against an object in sports (6.4 per 100,000 population) and pedal cycle riding (6.2 per 100,000). Ten to 19 year olds had the highest rate of sports and recreation related injury hospitalization (65.3 per 100,000 population), and males overall had a rate four times higher than females. CONCLUSIONS: Over 1700 sports and recreation related injuries occurred in Wisconsin in 2000 that were treated during an inpatient hospitalization. Sports and recreation activities result in a substantial number of serious, as well as minor injuries. Prevention efforts aimed at reducing injuries while continuing to promote participation in physical activity for all ages are critical. PMID- 15805438 TI - Assessing automobile head restraint positioning in Portland, Oregon. AB - OBJECTIVE: Automobile head restraints, when used properly, have been shown to decrease the incidence and severity of whiplash injuries to the neck. Before the development of a public campaign on proper head restraint positioning, the authors assessed head restraint positioning and public understanding. DESIGN: Over a one month period, the position of the vehicle head restraint of drivers was observed in moving cars in the city of Portland, Oregon (population 530,000). Optimal position was defined as having the head restraint above the ears with the back of the head touching the head restraint. A questionnaire on head restraint understanding was administered to people during jury service. RESULTS: Of the 4287 drivers observed, 1% (n = 30) had no head restraint on their seat, 4% (n = 158) had a fixed head restraint, and 95% (n = 4099) had an adjustable head restraint. Among the fixed head restraints, 21% (33/158) were positioned optimally with no horizontal gap. Among the adjustable head restraints, only 7% (280/4099) had optimal head restraint positioning. Overall, 93% (3974/4287) of all head restraints observed were suboptimally positioned. Seventy five percent (38/51) of polled Portland residents identified safety as the primary head restraint function. CONCLUSION: Ninety three percent of all head restraints observed were suboptimally positioned. Fixed head restraints were three times more likely to be in optimal position than adjustable head restraints (21% v 7%). Most polled Portland residents understood the proper function and positioning of head restraints. This discrepancy between actual practice and understanding should be addressed with public education and manufacturer design changes. PMID- 15805439 TI - A cost benefit analysis of an enhanced seat belt enforcement program in South Africa. AB - OBJECTIVE: To examine whether a program to increase the wearing of seat belts in a South African urban area would be worthwhile in societal terms. DESIGN: A cost benefit analysis of a one year enhanced seat belt enforcement program in eThekwini (Durban) Municipality. METHODS: Data were drawn from two main sources- a 1998 study of the cost of road crashes in South Africa and, given the absence of other data, a meta-analysis of the effectiveness of various types of interventions to reduce road crash casualties in the United States--and were analyzed using cost benefit analysis. RESULTS: A program designed to enforce greater wearing of seat belts, estimated to cost 2 million rand in one year, could be reasonably expected to increase seat belt usage rates by 16 percentage points and reduce fatalities and injuries by 9.5%. This would result in saved social costs of 13.6 million rand in the following year or a net present value of 11.6 million rand. There would also be favorable consequences for municipal finances. CONCLUSIONS: Investment in a program to increase seat belt wearing rates is highly profitable in societal terms. PMID- 15805440 TI - Long term effects of a home visit to prevent childhood injury: three year follow up of a randomized trial. AB - OBJECTIVE: To assess the long term effect of a home safety visit on the rate of home injury. DESIGN: Telephone survey conducted 36 months after participation in a randomized controlled trial of a home safety intervention. A structured interview assessed participant knowledge, beliefs, or practices around injury prevention and the number of injuries requiring medical attention. SETTING: Five pediatric teaching hospitals in four Canadian urban centres. PARTICIPANTS: Children less than 8 years of age presenting to an emergency department with a targeted home injury (fall, scald, burn, poisoning or ingestion, choking, or head injury while riding a bicycle), a non-targeted injury, or a medical illness. RESULTS: We contacted 774 (66%) of the 1172 original participants. A higher proportion of participants in the intervention group (63%) reported that home visits changed their knowledge, beliefs, or practices around the prevention of home injuries compared with those in the non-intervention group (43%; p<0.001). Over the 36 month follow up period the rate of injury visits to the doctor was significantly less for the intervention group (rate ratio = 0.74; 95% CI 0.63 to 0.87), consistent with the original (12 month) study results (rate ratio = 0.69; 95% CI 0.54 to 0.88). However, the effectiveness of the intervention appears to be diminishing with time (rate ratio for the 12-36 month study interval = 0.80; 95% CI 0.64 to 1.00). CONCLUSIONS: A home safety visit was able to demonstrate sustained, but modest, effectiveness of an intervention aimed at improving home safety and reducing injury. This study reinforces the need of home safety programs to focus on passive intervention and a simple well defined message. PMID- 15805441 TI - Working conditions and fatigue in professional truck drivers at Israeli ports. AB - BACKGROUND: Trucks represent 6% of all vehicles, but truck crashes account for 20% of road deaths in Israel, even though travel distances are usually short (<200 km) and overnight travel is uncommon. OBJECTIVE: To determine occupational and individual predictors of fatigue, falling asleep at the wheel, and involvement in crashes with injuries and deaths in truck drivers. SETTING AND METHODS: We carried out field interviews of 160 port truck drivers regarding driver characteristics, workplace and driving conditions, employer-employee relations, medical conditions, sleep quality and fatigue, falling asleep at the wheel, and involvement in road crashes. RESULTS: One day before interview, 38.1% of the drivers had worked more than the 12 hour legal limit. More than 30% reported falling asleep at the wheel recently, and 13% had prior involvement in a sleep related crash. Sixty seven (41.9%) drivers said that their employer forced them to work beyond the legal 12 hour daily limit. Involvement in a crash with casualties was associated with poor sleep quality (adjusted OR = 2.9; p = 0.042) and frequent difficulty finding parking when tired (OR = 3.7; p = 0.049). Self assessment of fatigue underestimated fatigue from the Pittsburgh Sleep Quality Questionnaire. However fatigue occurred in many drivers without sleep problems and many crashes occurred without fatigue. CONCLUSIONS: Prevention requires measures to reduce work stresses, screening drivers, speed control, and modal shifts. The work risks and adverse outcomes of truck drivers in large countries with long overnight journeys occur in a small country with small distances, relatively short work journeys, and little overnight travel. PMID- 15805442 TI - Gender differences for non-fatal unintentional fall related injuries among older adults. AB - OBJECTIVES: To quantify gender differences for non-fatal unintentional fall related injuries among US adults age 65 years and older treated in hospital emergency departments (EDs). METHODS: The authors analyzed data from a nationally representative sample of ED visits for January 2001 through December 2001, available through the National Electronic Injury Surveillance System All Injury Program (NEISS-AIP). For each initial ED visit, coders record one principal diagnosis (usually the most severe) and one primary part of the body affected. RESULTS: Based on 22,560 cases, an estimated 1.64 million older adults were treated in EDs for unintentional fall injuries. Of these, approximately 1.16 million, or 70.5%, were women. Fractures, contusions/abrasions, and lacerations accounted for more than three quarters of all injuries. Rates for injury diagnoses were generally higher among women, most notably for fractures which were 2.2 times higher than for men. For all parts of the body, women's injury rates exceeded those of men. Rate ratios were greatest for injuries of the leg/foot (2.3), arm/hand (2.0), and lower trunk (2.0). The hospitalization rate for women was 1.8 times that for men. CONCLUSIONS: Among older adults, non-fatal fall related injuries disproportionately affected women. Much is known about effective fall prevention strategies. We need to refine, promote, and implement these interventions. Additional research is needed to tailor interventions for different populations and to determine gender differences in the underlying causes and/or circumstances of falls. This information is vital for developing and implementing targeted fall prevention strategies. PMID- 15805443 TI - Out on a limb: risk factors for arm fracture in playground equipment falls. AB - OBJECTIVES: To investigate and quantify fall height, surface depth, and surface impact attenuation as risk factors for arm fracture in children who fall from playground equipment. DESIGN: Unmatched case control study. SETTING: Five case hospitals and 78 randomly selected control schools. PARTICIPANTS: Children aged less than 13 years in Victoria, Australia who fell from school playground equipment and landed on their arm. Cases sustained an upper limb fracture and controls had minor or no injury. A total of 402 cases and 283 controls were included. INTERVENTIONS: Children were interviewed in the playground as soon as possible after their fall. MAIN OUTCOME MEASURES: Falls were recreated on site using two validated impact test devices: a headform (measuring peak G and HIC) and a novel anthropometric arm load dummy. Equipment and fall heights, as well as surface depth and substrate were measured. RESULTS: Arm fracture risk was greatest for critical equipment heights above 1.5 m (OR 2.39, 95% CI 1.49 to 3.84, p<0.01), and critical fall heights above 1.0 m (OR 2.96, 95% CI 1.71 to 5.15, p<0.01). Peak headform deceleration below 100G was protective (OR 0.67, 95% CI 0.45 to 0.99, p = 0.04). Compliance with 20 cm surface depth recommendation was poor for both cases and controls. CONCLUSIONS: Arm fracture-specific criteria should be considered for future standards. These include surface and height conditions where critical headform deceleration is less than 100G. Consideration should also be given to reducing maximum equipment height to 1.5 m. Improved surface depth compliance and, in particular, guidelines for surface maintenance are required. PMID- 15805444 TI - Time trends in socioeconomic inequalities in road traffic injuries to children, Northumberland and Tyne and Wear 1988-2003. PMID- 15805445 TI - Prices and affordability in child restraint seats in Japan. PMID- 15805446 TI - Will the future continue to repeat the past? PMID- 15805447 TI - Can quality improvement be used to change the wider healthcare system? PMID- 15805448 TI - Who is responsible for the safety of infusion devices? It's high time for action! PMID- 15805449 TI - Impact of reporting hospital performance. PMID- 15805450 TI - Are the risks of hospital practice adequately recognised by incident reporting? PMID- 15805451 TI - Insights from the sharp end of intravenous medication errors: implications for infusion pump technology. AB - BACKGROUND: Intravenous (IV) medication errors are a common type of error identified in hospitals and can lead to considerable harm. Over the past 20 years there have been several hundred FDA reported incidents involving IV pumps, many of which have led to patient deaths. OBJECTIVE: To determine the actual types, frequency, and severity of medication errors associated with IV pumps. To evaluate the likelihood that smart pump technology without an interface to other systems could have prevented errors. METHODS: Using a point prevalence approach, investigators prospectively compared the medication, dose, and infusion rate on the IV pump with the prescribed medication, doses, and rate in the medical record. Preventability with smart pump technology was retrospectively determined based on a rigorous definition of currently available technology. RESULTS: A total of 426 medications were observed infusing through an IV pump. Of these, 285 (66.9%) had one or more errors associated with their administration. There were 389 documented errors overall; 37 were "rate deviation" errors and three of these were judged to be due to a programming mistake. Most of the documented events would not have caused patient harm (NCC MERP category C). Only one error would have been prevented by smart pump technology without additional interface and software capabilities. CONCLUSION: Medication errors associated with IV pumps occur frequently, have the potential to cause harm, and are epidemiologically diverse. Smart pumps are a necessary component of a comprehensive safe medication system. However, currently available smart pumps will fail to generate meaningful improvements in patient safety until they can be interfaced with other systems such as the electronic medical record, computerized prescriber order entry, bar coded medication administration systems, and pharmacy information systems. Future research should focus on the effectiveness of new technology in preventing latent and active errors, and on new types of error that any technology can introduce. PMID- 15805452 TI - Relationship between accreditation scores and the public disclosure of accreditation reports: a cross sectional study. AB - OBJECTIVE: To examine the association between accreditation scores and the disclosure of accreditation reports. DESIGN: A cross sectional study. SETTING: Hospitals participating in an accreditation programme in Japan. PARTICIPANTS: 547 of the 817 hospitals accredited by the Japan Council for Quality Health Care (JCQHC) by January 2003. MAIN OUTCOME MEASURES: Data on participation in public disclosure of accreditation reports through the JCQHC website were obtained from the JCQHC database. Comments on the disclosure were obtained using a questionnaire based survey. RESULTS: A total of 508 (93%) of the participating hospitals disclosed their accreditation reports on the JCQHC website. Public hospitals were significantly more committed to public disclosure than private hospitals, and larger hospitals were significantly more likely to participate in public disclosure than smaller hospitals. Accreditation scores were positively related to the public disclosure of hospital accreditation reports. Scores for patient focused care and efforts to meet community needs were significantly higher in actively disclosing hospitals than in non-disclosing hospitals. Among the large hospitals, scores for safety management were significantly higher in hospitals advocating disclosure than in non-disclosing hospitals. CONCLUSIONS: There was a positive correlation between accreditation scores and public disclosure. Our results suggest that the public disclosure of accreditation reports should be encouraged to improve public accountability and the quality of care. Future studies should investigate the interaction between public disclosure, processes and outcomes. PMID- 15805453 TI - Use of a systematic risk analysis method to improve safety in the production of paediatric parenteral nutrition solutions. AB - BACKGROUND: Until recently, the preparation of paediatric parenteral nutrition formulations in our institution included re-transcription and manual compounding of the mixture. Although no significant clinical problems have occurred, re engineering of this high risk activity was undertaken to improve its safety. Several changes have been implemented including new prescription software, direct recording on a server, automatic printing of the labels, and creation of a file used to pilot a BAXA MM 12 automatic compounder. The objectives of this study were to compare the risks associated with the old and new processes, to quantify the improved safety with the new process, and to identify the major residual risks. METHODS: A failure modes, effects, and criticality analysis (FMECA) was performed by a multidisciplinary team. A cause-effect diagram was built, the failure modes were defined, and the criticality index (CI) was determined for each of them on the basis of the likelihood of occurrence, the severity of the potential effect, and the detection probability. The CIs for each failure mode were compared for the old and new processes and the risk reduction was quantified. RESULTS: The sum of the CIs of all 18 identified failure modes was 3415 for the old process and 1397 for the new (reduction of 59%). The new process reduced the CIs of the different failure modes by a mean factor of 7. The CI was smaller with the new process for 15 failure modes, unchanged for two, and slightly increased for one. The greatest reduction (by a factor of 36) concerned re-transcription errors, followed by readability problems (by a factor of 30) and chemical cross contamination (by a factor of 10). The most critical steps in the new process were labelling mistakes (CI 315, maximum 810), failure to detect a dosage or product mistake (CI 288), failure to detect a typing error during the prescription (CI 175), and microbial contamination (CI 126). CONCLUSIONS: Modification of the process resulted in a significant risk reduction as shown by risk analysis. Residual failure opportunities were also quantified, allowing additional actions to be taken to reduce the risk of labelling mistakes. This study illustrates the usefulness of prospective risk analysis methods in healthcare processes. More systematic use of risk analysis is needed to guide continuous safety improvement of high risk activities. PMID- 15805454 TI - Pursuing integration of performance measures into electronic medical records: beta-adrenergic receptor antagonist medications. AB - OBJECTIVE: Electronic medical records seldom integrate performance indicators into daily operations. Assessing quality indicators traditionally requires resource intensive chart reviews of small samples. We sought to use an electronic medical record to assess use of beta-adrenergic antagonist medications (beta blockers) following myocardial infarction, to compare a standardized manual assessment with assessment using electronic medical records, and to discuss potential for future integration of performance indicators into electronic records. DESIGN: Cross-sectional data analysis. SETTING: An urban academic medical center. PARTICIPANTS: US Medicare beneficiaries 65 years of age or older, admitted to hospital with myocardial infarction between 1995 and 1999. MEASUREMENTS AND MAIN RESULTS: Manual chart review was compared with a computer driven assessment of electronic records. Administration of beta-blockers and cases excluded from use of beta-blockers were measured, based on Medicare criteria. Among 4490 older adults, 391 (4%) of 9018 hospital admissions contained codes for myocardial infarction. In 323 (83%) of the 391 hospital admissions, criteria for excluding beta-blockers were met; 235 (60%) were excluded due to heart failure. Of 68 hospital admissions for myocardial infarction that did not meet exclusion criteria, physicians prescribed beta-blockers in 49 (72%) on admission and 42 (62%) at discharge. Compared with manual chart review, electronic review had a sensitivity of 83-100% and led to fewer false negative findings. CONCLUSIONS: An electronic medical records system can be used instead of chart review to measure use of beta-blockers after myocardial infarction. This should lead to integration of real time automated performance measurement into electronic medical records. PMID- 15805456 TI - Do split-side rails present an increased risk to patient safety? AB - BACKGROUND: Concerns have been raised about the safety of split-side bed rails for patients in the UK. OBJECTIVES: To investigate whether split-side rails were more likely to be associated with entrapment and injury of patients than other bed rail types. To establish whether there was a difference in the site of injury caused by different bed rail types and whether the outcome of the injury (death versus survival) varied by bedrail type. METHODS: A search of the USA Food and Drug Administration MAUDE database was carried out. The reports were screened using rigorous inclusion/exclusion criteria and then coded for rail type, incident outcome, and area of body involved. RESULTS: Split-side rail incidents only accounted for 5% of the reports and were more likely to involve the chest or pelvis. Although the biggest overall risk by rail type cannot be determined from these data, the severity of the outcome changed with the equipment type. Incidents involving half rails were more likely to be associated with head, neck, or face entrapments and were also more likely than other bed rail types to result in death. DISCUSSION: Split-side rail entrapments were not a common occurrence. However, our findings suggest that bed rails are associated with some level of risk of entrapment that potentially could result in death. Healthcare providers should therefore ensure that they follow the guidelines for risk assessment and rail use from the MHRA and other professional bodies so that the cultural norm in the UK continues to be "opt in", where no bed rails are used unless indicated by a documented clinical assessment. PMID- 15805455 TI - Effect on the process of care of an active strategy to implement clinical guidelines on physiotherapy for low back pain: a cluster randomised controlled trial. AB - OBJECTIVES: To evaluate the effect on the process of care of an active strategy to implement clinical guidelines on physiotherapy for low back pain. DESIGN: A cluster randomised controlled trial comparing an active strategy with standard dissemination. SETTING: Primary care physiotherapy practices. PARTICIPANTS: 113 physiotherapists were randomly allocated to receive the guidelines by mail (control group) or to receive an additional active strategy (intervention group) which consisted of a multifaceted programme including education, discussion, role playing, feedback, and reminders. MAIN OUTCOME MEASURES: Adherence to the guidelines was measured by means of individual patients' forms recording the treatment completed by the physiotherapist. The forms were assessed using an algorithm based on the number of treatment sessions, treatment goals, interventions, and patient education. RESULTS: Physiotherapists in the intervention group more often correctly limited the number of treatment sessions for patients with a normal course of back pain (OR 2.39; 95% CI 1.12 to 5.12), more often set functional treatment goals (OR 1.99; 95% CI 1.06 to 3.72), more often used mainly active interventions (OR 2.79; 95% CI 1.19 to 6.55), and more often gave adequate patient education (OR 3.59; 95% CI 1.35 to 9.55). They also adhered more to all four criteria (OR 2.05; 95% CI 1.15 to 3.65). CONCLUSIONS: The active strategy moderately improved adherence to the guidelines. Active strategies are recommended to implement the clinical guidelines on physiotherapy for low back pain. PMID- 15805458 TI - A web-based incident reporting system and multidisciplinary collaborative projects for patient safety in a Japanese hospital. AB - PROBLEM: When patient safety programs were mandated for Japanese health care institutions, a safety culture, a tool for collecting incident reports, an organizational arrangement for multidisciplinary collaboration, and interventional methods for improvement had to be established. DESIGN: Observational study of effects of new patient safety programs. SETTING: Osaka University Hospital, a large government-run teaching hospital. STRATEGY FOR CHANGE: A voluntary and anonymous web-based incident reporting system was introduced. For the new organizational structure a clinical risk management committee, a department of clinical quality management, and area clinical risk managers were established with their respective roles clearly defined to advance the plan-do-study-act cycle and to integrate efforts. For preventive action, alert procedures, staff education, ward rounds by peers, a system oriented approach for reducing errors, and various feedback channels were introduced. EFFECTS OF CHANGE: Continuous incident reporting by all hospital staff has been observed since the introduction of the new system. Several error inducing situations have been improved: wrong choice of drug in computer prescribing, maladministration of drugs due to a look-alike appearance or confusion about the manipulation of a medical device, and poor after hours service of the blood transfusion unit. Staff participation in educational seminars has been dramatically improved. Ward rounds have detected problematic procedures which needed to be dealt with. LESSONS LEARNT: Patient safety programs based on a web based incident reporting system, responsible persons, staff education, and a variety of feedback procedures can help promote a safety culture, multidisciplinary collaboration, and strong managerial leadership resulting in system oriented improvement. PMID- 15805457 TI - Relationship between tort claims and patient incident reports in the Veterans Health Administration. AB - OBJECTIVE: The Veterans Health Administration's patient incident reporting system was established to obtain comprehensive data on adverse events that affect patients and to act as a harbinger for risk management. It maintains a dataset of tort claims that are made against Veterans Administration's employees acting within the scope of employment. In an effort to understand the thoroughness of reporting, we examined the relationship between tort claims and patient incident reports (PIRs). METHODS: Using social security and record numbers, we matched 8260 tort claims and 32 207 PIRs from fiscal years 1993-2000. Tort claims and PIRs were considered to be related if the recorded dates of incident were within 1 month of each other. Descriptive statistics, odds ratios, and two sample t tests with unequal variances were used to determine the relationship between PIRs and tort claims. RESULTS: 4.15% of claims had a related PIR. Claim payment (either settlement or judgment for plaintiff) was more likely when associated with a PIR (OR 3.62; 95% CI 2.87 to 4.60). Payment was most likely for medication errors (OR 8.37; 95% CI 2.05 to 73.25) and least likely for suicides (OR 0.25; 95% CI 0.11 to 0.55). CONCLUSIONS: Although few tort claims had a related PIR, if a PIR was present the tort claim was more likely to result in a payment; moreover, the payment was likely to be higher. Underreporting of patient incidents that developed into tort claims was evident. Our findings suggest that, in the Veterans Health Administration, there is a higher propensity to both report and settle PIRs with bad outcomes. PMID- 15805459 TI - "Going solid": a model of system dynamics and consequences for patient safety. AB - Rather than being a static property of hospitals and other healthcare facilities, safety is dynamic and often on short time scales. In the past most healthcare delivery systems were loosely coupled-that is, activities and conditions in one part of the system had only limited effect on those elsewhere. Loose coupling allowed the system to buffer many conditions such as short term surges in demand. Modern management techniques and information systems have allowed facilities to reduce inefficiencies in operation. One side effect is the loss of buffers that previously accommodated demand surges. As a result, situations occur in which activities in one area of the hospital become critically dependent on seemingly insignificant events in seemingly distant areas. This tight coupling condition is called "going solid". Rasmussen's dynamic model of risk and safety can be used to formulate a model of patient safety dynamics that includes "going solid" and its consequences. Because the model addresses the dynamic aspects of safety, it is particularly suited to understanding current conditions in modern healthcare delivery and the way these conditions may lead to accidents. PMID- 15805460 TI - Implementing a national strategy for patient safety: lessons from the National Health Service in England. AB - Improving patient safety has become a core issue for many modern healthcare systems. However, knowledge of the best ways for government initiated efforts to improve patient safety is still evolving, although there is considerable commonality in the challenges faced by countries. Actions to improve patient safety must operate at multiple levels of the healthcare system simultaneously. Using the example of the NHS in England, this article highlights the importance of a strategic analysis of the policy process and the prevailing policy context in the design of the national patient safety strategy. The paper identifies a range of policy "levers" (forces for change) that can be used to support the implementation of the national safety initiative and, in particular, discusses the strengths and limitations of the "business case" approach that has attracted recent interest. The paper offers insights into the implementation of national patient safety goals that should provide learning for other countries. PMID- 15805461 TI - Pasteur and parachutes: when statistical process control is better than a randomized controlled trial. PMID- 15805462 TI - Effective health care: management of head and neck cancers. AB - The management of head and neck cancer, published in a recent issue of Effective Health Care, is reviewed. PMID- 15805463 TI - Kissing complex RNAs mediate interaction between the Fragile-X mental retardation protein KH2 domain and brain polyribosomes. AB - Fragile-X mental retardation is caused by loss of function of a single gene encoding the Fragile-X mental retardation protein, FMRP, an RNA-binding protein that harbors two KH-type and one RGG-type RNA-binding domains. Previous studies identified intramolecular G-quartet RNAs as high-affinity targets for the RGG box, but the relationship of RNA binding to FMRP function and mental retardation remains unclear. One severely affected patient harbors a missense mutation (I304N) within the second KH domain (KH2), and some evidence suggests this domain may be involved in the proposed role of FMRP in translational regulation. We now identify the RNA target for the KH2 domain as a sequence-specific element within a complex tertiary structure termed the FMRP kissing complex. We demonstrate that the association of FMRP with brain polyribosomes is abrogated by competition with the FMRP kissing complex RNA, but not by high-affinity G-quartet RNAs. We conclude that mental retardation associated with the I304N mutation, and likely the Fragile-X syndrome more generally, may relate to a crucial role for RNAs harboring the kissing complex motif as targets for FMRP translational regulation. PMID- 15805464 TI - An MT1-MMP-PDGF receptor-beta axis regulates mural cell investment of the microvasculature. AB - Platelet-derived growth factor (PDGF)/PDGFRbeta-dependent investment of the vascular endothelium by mural cells (i.e., pericytes and vascular smooth muscle cells; VSMCs) is critical for normal vessel wall structure and function. In the developing vasculature, mural cell recruitment is associated with the functionally undefined expression of the type I transmembrane proteinase, membrane-type 1 matrix metalloproteinase (MT1-MMP). In this paper, using VSMCs and tissues isolated from gene-targeted mice, we identify MT1-MMP as a PDGF-B selective regulator of PDGFRbeta-dependent signal transduction and mural cell function. In VSMCs, catalytically active MT1-MMP associates with PDGFRbeta in membrane complexes that support the efficient induction of mitogenic signaling by PDGF-B in a matrix metalloproteinase inhibitor-sensitive fashion. In contrast, MT1-MMP-deficient VSMCs display PDGF-B-selective defects in chemotaxis and proliferation as well as ERK1/2 and Akt activation that can be rescued in tandem fashion following retroviral transduction with the wild-type protease. Consistent with these in vitro findings, MT1-MMP-deficient brain tissues display a marked reduction in mural cell density as well as abnormal vessel wall morphology similar to that reported in mice expressing PDGF-B or PDGFRbeta hypomorphic alleles. Together, these data identify MT1-MMP as a novel proteolytic modifier of PDGF-B/PDGFRbeta signal transduction that cooperatively regulates vessel wall architecture in vivo. PMID- 15805466 TI - The nucleolus as a stress sensor: JNK2 inactivates the transcription factor TIF IA and down-regulates rRNA synthesis. AB - Cells respond to a variety of extracellular and intracellular forms of stress by down-regulating rRNA synthesis. We have investigated the mechanism underlying stress-dependent inhibition of RNA polymerase I (Pol I) transcription and show that the Pol I-specific transcription factor TIF-IA is inactivated upon stress. Inactivation is due to phosphorylation of TIF-IA by c-Jun N-terminal kinase (JNK) at a single threonine residue (Thr 200). Phosphorylation at Thr 200 impairs the interaction of TIF-IA with Pol I and the TBP-containing factor TIF-IB/SL1, thereby abrogating initiation complex formation. Moreover, TIF-IA is translocated from the nucleolus into the nucleoplasm. Substitution of Thr 200 by valine as well as knock-out of Jnk2 prevent inactivation and translocation of TIF-IA, leading to stress-resistance of Pol I transcription. Our data identify TIF-IA as a downstream target of the JNK pathway and suggest a critical role of JNK2 to protect rRNA synthesis against the harmful consequences of cellular stress. PMID- 15805467 TI - Visualizing Mu transposition: assembling the puzzle pieces. PMID- 15805465 TI - Replication checkpoint kinase Cds1 regulates Mus81 to preserve genome integrity during replication stress. AB - The replication checkpoint kinase Cds1 preserves genome integrity by stabilizing stalled replication forks. Cds1 targets substrates through its FHA domain. The Cds1 FHA domain interacts with Mus81, a subunit of the Mus81-Eme1 structure specific endonuclease. We report here that Mus81 and Rhp51 are required for generating deletion mutations in fission yeast replication mutants that experience replication stress. A mutation in the Mus81 FHA-binding motif eliminates its Cds1-binding and Cds1-dependent phosphorylation. Furthermore, this mutation exacerbates the deletion mutator phenotype of a replication mutant, and induces a hyper-recombination phenotype in hydroxyurea-treated cells. In unperturbed cells, Mus81 associates with chromatin throughout S phase. In replication mutants grown at semipermissive temperature, Mus81 undergoes minor Cds1-dependent phosphorylation, remains chromatin-associated, generates deletion mutations, and maintains cell growth. Upon S-phase arrest by acute hydroxyurea treatment, Mus81 is not required for cell viability but is essential for recovery from replication fork collapse. Moreover, Mus81 undergoes extensive Cds1 dependent phosphorylation and dissociates from chromatin in hydroxyurea-arrested cells, thereby preventing it from cleaving stalled replication forks that could lead to fork breakage and chromosomal rearrangement. These results provide novel insights into how Cds1 regulates Mus81 accordingly when cells experience different replication stress to preserve genome integrity. PMID- 15805468 TI - Ubiquitin ligase MKRN1 modulates telomere length homeostasis through a proteolysis of hTERT. AB - Telomere homeostasis is regulated by telomerase and a collection of associated proteins. Telomerase is, in turn, regulated by post-translational modifications of the rate-limiting catalytic subunit hTERT. Here we show that disruption of Hsp90 by geldanamycin promotes efficient ubiquitination and proteasome-mediated degradation of hTERT. Furthermore, we have used the yeast two-hybrid method to identify a novel RING finger gene (MKRN1) encoding an E3 ligase that mediates ubiquitination of hTERT. Overexpression of MKRN1 in telomerase-positive cells promotes the degradation of hTERT and decreases telomerase activity and subsequently telomere length. Our data suggest that MKRN1 plays an important role in modulating telomere length homeostasis through a dynamic balance involving hTERT protein stability. PMID- 15805469 TI - A novel cell-to-cell trafficking assay indicates that the KNOX homeodomain is necessary and sufficient for intercellular protein and mRNA trafficking. AB - Cell-to-cell trafficking of regulatory proteins is a novel mechanism for communication during cell fate specification in plants. Although several developmental proteins traffic cell-to-cell, no signals that are both necessary and sufficient for this function in developmental proteins have been described. We developed a novel trafficking assay using trichome rescue in Arabidopsis. Fusion to KNOTTED1 (KN1) conferred gain-of-trafficking function to the cell autonomous GLABROUS1 (GL1) protein. We show that the KNOX homeodomain (HD) is necessary and sufficient for intercellular trafficking, identifying a novel function for the HD as the minimal sequence required for trafficking of KN1 and its associated mRNA. PMID- 15805470 TI - Histone deacetylase 3 (HDAC3) activity is regulated by interaction with protein serine/threonine phosphatase 4. AB - Histone deacetylase 3 (HDAC3) is one of four members of the human class I HDACs that regulates gene expression by deacetylation of histones and nonhistone proteins. Early studies have suggested that HDAC3 activity is regulated by association with the corepressors N-CoR and SMRT. Here we demonstrate that, in addition to protein-protein interactions with NCoR/SMRT, the activity of HDAC3 is regulated by both phosphorylation and dephosphorylation. A protein kinase CK2 phosphoacceptor site in the HDAC3 protein was identified at position Ser424, which is a nonconserved residue among the class I HDACs. Mutation of this residue was found to reduce deacetylase activity. Interestingly, unlike other class I HDACs, HDAC3 uniquely copurifies with the catalytic and regulatory subunits of the protein serine/threonine phosphatase 4 complex (PP4c/PP4R1). Furthermore, HDAC3 complexes displayed protein phosphatase activity and a series of subsequent mutational analyses revealed that the N terminus of HDAC3 (residues 1-122) was both necessary and sufficient for HDAC3-PP4c interactions. Significantly, both overexpression and siRNA knock-down approaches, and analysis of cells devoid of PP4c, unequivocally show that HDAC3 activity is inversely proportional to the cellular abundance of PP4(c). These findings therefore further highlight the importance of protein-protein interactions and extend the significance of dephosphorylation in the regulation of HDAC activity, as well as present a novel alternative pathway by which HDAC3 activity is regulated. PMID- 15805471 TI - Ddb1 controls genome stability and meiosis in fission yeast. AB - The human UV-damaged DNA-binding protein Ddb1 associates with cullin 4 ubiquitin ligases implicated in nucleotide excision repair (NER). These complexes also contain the signalosome (CSN), but NER-relevant ubiquitination targets have not yet been identified. We report that fission yeast Ddb1, Cullin 4 (Pcu4), and CSN subunits Csn1 and Csn2 are required for degradation of the ribonucleotide reductase (RNR) inhibitor protein Spd1. Ddb1-deficient cells have >20-fold increased spontaneous mutation rate. This is partly dependent on the error-prone translesion DNA polymerases. Spd1 deletion substantially reduced the mutation rate, suggesting that insufficient RNR activity accounts for approximately 50% of observed mutations. Epistasis analysis indicated that Ddb1 contributed to mutation avoidance and tolerance to DNA damage in a pathway distinct from NER. Finally, we show that Ddb1/Csn1/Cullin 4-mediated Spd1 degradation becomes essential when cells differentiate into meiosis. These results suggest that Ddb1, along with Cullin 4 and the signalosome, constitute a major pathway controlling genome stability, repair, and differentiation via RNR regulation. PMID- 15805472 TI - Multiple branches of the meiotic recombination pathway contribute independently to homolog pairing and stable juxtaposition during meiosis in budding yeast. AB - A unique aspect of meiosis is the segregation of homologous chromosomes at the meiosis I division. Homologs are physically connected prior to segregation by crossing over between nonsister chromatids. Crossovers arise from the repair of induced double-strand breaks (DSBs). In many organisms, more DSBs are formed than crossovers in a given nucleus. It has been previously suggested that repair of DSBs to noncrossover recombination products aids homolog alignment. Here we explore how two modes of the meiotic recombination pathway (crossover and noncrossover) and meiotic telomere reorganization contribute to the pairing and close juxtaposition of homologous chromosomes in budding yeast. We found that intermediates in the DSB repair pathway leading to both crossover and noncrossover recombination products contribute independently to close, stable homolog juxtaposition (CSHJ), a measurable state of homolog pairing. Analysis of the ndj1delta mutant indicates that the effect of meiotic telomere reorganization on CSHJ is exerted through recombination intermediates at interstitial chromosomal loci, perhaps through the noncrossover branch of the DSB repair pathway. We suggest that transient, early DSB-initiated interactions, including those that give rise to noncrossovers, are important for homolog recognition and juxtaposition. PMID- 15805473 TI - Virus induction of heat shock protein 70 reflects a general response to protein accumulation in the plant cytosol. AB - Different cytoplasmically replicating RNA viruses were shown to induce a specific subset of heat-inducible heat shock protein 70 (HSP70) genes in Arabidopsis (Arabidopsis thaliana). To identify the inducing principle, a promoterreporter system was developed for the facile analysis of differentially responding Arabidopsis HSP70 genes, by infiltration into Nicotiana benthamiana leaves. Through transient expression of individual viral cistrons or through deletion analysis of a viral replicon, we were unable to identify a unique inducer of HSP70. However, there was a positive correlation between the translatability of the test construct and the differential induction of HSP70. Since these data implied a lack of specificity in the induction process, we also expressed a random series of cytosolically targeted Arabidopsis genes and showed that these also differentially induced HSP70. Through a comparison of different promoterreporter constructs and through measurements of the steady-state levels of the individual proteins, it appeared that the HSP70 response reflected the ability of the cytosol to sense individual properties of particular proteins when expressed at high levels. This phenomenon is reminiscent of the unfolded protein response observed when the induced accumulation of proteins in the endoplasmic reticulum also induces a specific suite of chaperones. PMID- 15805474 TI - Antifreeze proteins modify the freezing process in planta. AB - During cold acclimation, winter rye (Secale cereale L. cv Musketeer) plants accumulate antifreeze proteins (AFPs) in the apoplast of leaves and crowns. The goal of this study was to determine whether these AFPs influence survival at subzero temperatures by modifying the freezing process or by acting as cryoprotectants. In order to inhibit the growth of ice, AFPs must be mobile so that they can bind to specific sites on the ice crystal lattice. Guttate obtained from cold-acclimated winter rye leaves exhibited antifreeze activity, indicating that the AFPs are free in solution. Infrared video thermography was used to observe freezing in winter rye leaves. In the absence of an ice nucleator, AFPs had no effect on the supercooling temperature of the leaves. However, in the presence of an ice nucleator, AFPs lowered the temperature at which the leaves froze by 0.3 degrees C to 1.2 degrees C. In vitro studies showed that apoplastic proteins extracted from cold-acclimated winter rye leaves inhibited the recrystallization of ice and also slowed the rate of migration of ice through solution-saturated filter paper. When we examined the possible role of winter rye AFPs in cryoprotection, we found that lactate dehydrogenase activity was higher after freezing in the presence of AFPs compared with buffer, but the same effect was obtained by adding bovine serum albumin. AFPs had no effect on unstacked thylakoid volume after freezing, but did inhibit stacking of the thylakoids, thus indicating a loss of thylakoid function. We conclude that rye AFPs have no specific cryoprotective activity; rather, they interact directly with ice in planta and reduce freezing injury by slowing the growth and recrystallization of ice. PMID- 15805475 TI - Betaxanthins as substrates for tyrosinase. An approach to the role of tyrosinase in the biosynthetic pathway of betalains. AB - Tyrosinase or polyphenol oxidase (EC 1.14.18.1) is the key enzyme in melanin biosynthesis and in the enzymatic browning of fruits and vegetables. The role of tyrosinase in the secondary metabolism of plants still remains unclear, but its implication in betalain biosynthesis has been proposed. Betalains are an important class of water-soluble pigments, characteristic of plants belonging to the order Caryophyllales. In this article, the betaxanthins, tyrosine-betaxanthin (portulacaxanthin II) and dopaxanthin, are reported to be physiological substrates for tyrosinase. The direct activity of tyrosinase on selected betaxanthins is characterized in depth, and conversion of tyrosine-betaxanthin to dopaxanthin and its further oxidation to a series of compounds are described. Identity of the reaction products was studied by high-performance liquid chromatography and electrospray ionization-mass spectrometry. Masses determined for the reaction products were the same in all cases, 389 m/z ([M + H]+) and equal to that determined for betanidin. Data indicate that dopaxanthin-quinone is obtained and evolves to more stable species by intramolecular cyclization. Kinetic parameters for tyrosinase acting on dopaxanthin were evaluated, showing a high affinity for this substrate (K(m) = 84.3 microM). The biosynthetic scheme of betalains is reviewed and a branch is proposed based on the description of physiological substrates for tyrosinase. Lampranthus productus, Glottiphylum oligocarpum, and Glottiphylum pigmaeum are described as sources of stereopure (2S/S)-dopaxanthin. PMID- 15805476 TI - O-acetylserine and the regulation of expression of genes encoding components for sulfate uptake and assimilation in potato. AB - cDNAs encoding a high-affinity sulfate transporter and an adenosine 5' phosphosulfate reductase from potato (Solanum tuberosum L. cv Desiree) have been cloned and used to examine the hypothesis that sulfate uptake and assimilation is transcriptionally regulated and that this is mediated via intracellular O acetylserine (OAS) pools. Gas chromotography coupled to mass spectrometry was used to quantify OAS and its derivative, N-acetylserine. Treatment with external OAS increased sulfate transporter and adenosine 5'-phosphosulfate reductase gene expression consistent with a model of transcriptional induction by OAS. To investigate this further, the Escherichia coli gene cysE (serine acetyltransferase EC 2.3.1.30), which synthesizes OAS, has been expressed in potato to modify internal metabolite pools. Transgenic lines, with increased cysteine and glutathione pools, particularly in the leaves, had increased sulfate transporter expression in the roots. However, the small increases in the OAS pools were not supportive of the hypothesis that this molecule is the signal of sulfur (S) nutritional status. In addition, although during S starvation the content of S-containing compounds decreased (consistent with derepression as a mechanism of regulation), OAS pools increased only following extended starvation, probably as a consequence of the S starvation. Taken together, expression of these genes may be induced by a demand-driven model, via a signal from the shoots, which is not OAS. Rather, the signal may be the depletion of intermediates of the sulfate assimilation pathway, such as sulfide, in the roots. Finally, sulfate transporter activity did not increase in parallel with transcript and protein abundance, indicating additional posttranslational regulatory mechanisms. PMID- 15805477 TI - Comprehensive interaction map of the Arabidopsis MADS Box transcription factors. AB - Interactions between proteins are essential for their functioning and the biological processes they control. The elucidation of interaction maps based on yeast studies is a first step toward the understanding of molecular networks and provides a framework of proteins that possess the capacity and specificity to interact. Here, we present a comprehensive plant protein-protein interactome map of nearly all members of the Arabidopsis thaliana MADS box transcription factor family. A matrix-based yeast two-hybrid screen of >100 members of this family revealed a collection of specific heterodimers and a few homodimers. Clustering of proteins with similar interaction patterns pinpoints proteins involved in the same developmental program and provides valuable information about the participation of uncharacterized proteins in these programs. Furthermore, a model is proposed that integrates the floral induction and floral organ formation networks based on the interactions between the proteins involved. Heterodimers between flower induction and floral organ identity proteins were observed, which point to (auto)regulatory mechanisms that prevent the activity of flower induction proteins in the flower. PMID- 15805478 TI - Cloning and characterization of microRNAs from rice. AB - MicroRNAs (miRNAs) are a growing family of small noncoding RNAs that downregulate gene expression in a sequence-specific manner. The identification of the entire set of miRNAs from a model organism is a critical step toward understanding miRNA guided gene regulation. Rice (Oryza sativa) and Arabidopsis thaliana, two plant model species with fully sequenced genomes, are representatives of monocotyledonous and dicotyledonous flowering plants, respectively. Thus far, experimental identification of miRNAs in plants has been confined to Arabidopsis. Computational analysis based on conservation with known miRNAs from Arabidopsis has predicted 20 families of miRNAs in rice. To identify miRNAs that are difficult to predict in silico or not conserved in Arabidopsis, we generated three cDNA libraries of small RNAs from rice shoot, root, and inflorescence tissues. We identified 35 miRNAs, of which 14 are new, and these define 13 new families. Thirteen of the new miRNAs are not conserved in Arabidopsis. Four of the new miRNAs are conserved in related monocot species but not in Arabidopsis, which suggests that these may have evolved after the divergence of monocots and dicots. The remaining nine new miRNAs appear to be absent in the known sequences of other plant species. Most of the rice miRNAs are expressed ubiquitously in all tissues examined, whereas a few display tissue-specific expression. We predicted 46 genes as targets of the new rice miRNAs: 16 of these predicted targets encode transcription factors, and other target genes appear to play roles in diverse physiological processes. Four target genes have been experimentally verified by detection of miRNA-mediated mRNA cleavage. Our identification of new miRNAs in rice suggests that these miRNAs may have evolved independently in rice or been lost in other species. PMID- 15805479 TI - DDM1 binds Arabidopsis methyl-CpG binding domain proteins and affects their subnuclear localization. AB - Methyl-CpG binding domain (MBD) proteins in Arabidopsis thaliana bind in vitro methylated CpG sites. Here, we aimed to characterize the binding properties of AtMBDs to chromatin in Arabidopsis nuclei. By expressing in wild-type cells AtMBDs fused to green fluorescent protein (GFP), we showed that AtMBD7 was evenly distributed at all chromocenters, whereas AtMBD5 and 6 showed preference for two perinucleolar chromocenters adjacent to nucleolar organizing regions. AtMBD2, previously shown to be incapable of binding in vitro-methylated CpG, was dispersed within the nucleus, excluding chromocenters and the nucleolus. Recruitment of AtMBD5, 6, and 7 to chromocenters was disrupted in ddm1 and met1 mutant cells, where a significant reduction in cytosine methylation occurs. In these mutant cells, however, AtMBD2 accumulated at chromocenters. No effect on localization was observed in the chromomethylase3 mutant showing reduced CpNpG methylation or in kyp-2 displaying a reduction in Lys 9 histone H3 methylation. Transient expression of DDM1 fused to GFP showed that DDM1 shares common sites with AtMBD proteins. Glutathione S-transferase pull-down assays demonstrated that AtMBDs bind DDM1; the MBD motif was sufficient for this interaction. Our results suggest that the subnuclear localization of AtMBD is not solely dependent on CpG methylation; DDM1 may facilitate localization of AtMBDs at specific nuclear domains. PMID- 15805480 TI - The N-terminal domain of chlorophyllide a oxygenase confers protein instability in response to chlorophyll B accumulation in Arabidopsis. AB - Plants acclimate to variations in light intensity by changing the antenna size of photosystems. This acclimation allows them to undergo efficient photosynthesis and creates a protective strategy to minimize photodamage. Chlorophyll b synthesis by chlorophyllide a oxygenase (CAO) is a key regulatory step in the control of antenna size. Recently, we found that higher plant CAOs consist of three domains (A, B, and C domains) and confirmed that the C domain possesses catalytic function. To investigate the function of the A domain, we fused various combinations of these three domains with green fluorescent protein (GFP) and introduced them into Arabidopsis thaliana. When a full-length CAO-GFP fusion protein was introduced into a chlorophyll b-less chlorina1-1 mutant, chlorophyll b accumulated to almost the same levels as in the chlorophyll b-containing Columbia wild type, but the CAO-GFP could not be detected by immunoblotting. By contrast, when a GFP-C domain fusion was introduced into chlorina1-1 or Columbia wild type, a large amount of GFP-C domain protein accumulated and the chlorophyll a/b ratio decreased drastically from 3.6 to 2.2 in Columbia wild type. When an A domain-GFP was introduced into Columbia wild type, A domain-GFP levels were very low. Conversely, a large amount of the protein accumulated when it was introduced into the chlorina1-1 mutant. These results indicate that the A domain may sense the presence of chlorophyll b and regulate the accumulation of CAO protein in the chloroplasts. PMID- 15805481 TI - Mutation of SAC1, an Arabidopsis SAC domain phosphoinositide phosphatase, causes alterations in cell morphogenesis, cell wall synthesis, and actin organization. AB - SAC (for suppressor of actin) domain proteins in yeast and animals have been shown to modulate the levels of phosphoinositides, thereby regulating several cellular activities such as signal transduction, actin cytoskeleton organization, and vesicle trafficking. Nine genes encoding SAC domain-containing proteins are present in the Arabidopsis thaliana genome, but their roles in plant cellular functions and plant growth and development have not been characterized. In this report, we demonstrate the essential roles of one of the Arabidopsis SAC domain proteins, AtSAC1, in plant cellular functions. Mutation of the AtSAC1 gene in the fragile fiber7 (fra7) mutant caused a dramatic decrease in the wall thickness of fiber cells and vessel elements, thus resulting in a weak stem phenotype. The fra7 mutation also led to reduced length and aberrant shapes in fiber cells, pith cells, and trichomes and to an alteration in overall plant architecture. The AtSAC1 gene was found to be expressed in all tissues in elongating organs; however, it showed predominant expression in vascular tissues and fibers in nonelongating parts of stems. In vitro activity assay demonstrated that AtSAC1 exhibited phosphatase activity toward phosphatidylinositol 3,5-biphosphate. Subcellular localization studies showed that AtSAC1 was colocalized with a Golgi marker. Truncation of the C terminus by the fra7 mutation resulted in its localization in the cytoplasm but had no effect on phosphatase activity. Furthermore, examination of the cytoskeleton organization revealed that the fra7 mutation caused the formation of aberrant actin cables in elongating cells but had no effect on the organization of cortical microtubules. Together, these results provide genetic evidence that AtSAC1, a SAC domain phosphoinositide phosphatase, is required for normal cell morphogenesis, cell wall synthesis, and actin organization. PMID- 15805482 TI - Molecular and functional dissection of the maize B chromosome centromere. AB - The centromere of the maize (Zea mays) B chromosome contains several megabases of a B-specific repeat (ZmBs), a 156-bp satellite repeat (CentC), and centromere specific retrotransposons (CRM elements). Here, we demonstrate that only a small fraction of the ZmBs repeats interacts with CENH3, the histone H3 variant specific to centromeres. CentC, which marks the CENH3-associated chromatin in maize A centromeres, is restricted to an approximately 700-kb domain within the larger context of the ZmBs repeats. The breakpoints of five B centromere misdivision derivatives are mapped within this domain. In addition, the fraction of this domain remaining after misdivision correlates well with the quantity of CENH3 on the centromere. Thus, the functional boundaries of the B centromere are mapped to a relatively small CentC- and CRM-rich region that is embedded within multimegabase arrays of the ZmBs repeat. Our results demonstrate that the amount of CENH3 at the B centromere can be varied, but with decreasing amounts, the function of the centromere becomes impaired. PMID- 15805483 TI - Regulation by external K+ in a maize inward shaker channel targets transport activity in the high concentration range. AB - An inward Shaker K(+) channel identified in Zea mays (maize), ZmK2.1, displays strong regulation by external K(+) when expressed in Xenopus laevis (African clawed frog) oocytes or COS cells. ZmK2.1 is specifically activated by K(+) with an apparent K(m) close to 15 mM independent of the membrane hyperpolarization level. In the absence of K(+), ZmK2.1 appears to enter a nonconducting state. Thus, whatever the membrane potential, this maize channel cannot mediate K(+) influx in the submillimolar concentration range, unlike its relatives in Arabidopsis thaliana. Its expression is restricted to the shoots, the strongest signal (RT-PCR) being associated with vascular/bundle sheath strands. Based on sequence and gene structure, the closest relatives of ZmK2.1 in Arabidopsis are K(+) Arabidopsis Transporter 1 (KAT1) (expressed in guard cells) and KAT2 (expressed in guard cells and leaf phloem). Patch-clamp analyses of guard cell protoplasts reveal a higher functional diversity of K(+) channels in maize than in Arabidopsis. Channels endowed with regulation by external K(+) similar to that of ZmK2.1 (channel activity regulated by external K(+) with a K(m) close to 15 mM, regulation independent of external Ca(2+)) constitute a major component of the maize guard cell inward K(+) channel population. The presence of such channels in maize might reflect physiological traits of C4 and/or monocotyledonous plants. PMID- 15805484 TI - BLADE-ON-PETIOLE-dependent signaling controls leaf and floral patterning in Arabidopsis. AB - NONEXPRESSOR OF PR GENES1 (NPR1) is a key regulator of the plant defense response known as systemic acquired resistance. Accumulation of the signal molecule salicylic acid (SA) leads to a change in intracellular redox potential, enabling NPR1 to enter the nucleus and interact with TGACG sequence-specific binding protein (TGA) transcription factors, which in turn bind to SA-responsive elements in the promoters of defense genes. Here, we show that two NPR1-like genes, BLADE ON-PETIOLE1 (BOP1) and BOP2, function redundantly to control growth asymmetry, an important aspect of patterning in leaves and flowers. Phenotypes in the double mutant include leafy petioles, loss of floral organ abscission, and asymmetric flowers subtended by a bract. We demonstrate that BOP2 is localized to both the nucleus and the cytoplasm, but unlike NPR1, it is highly expressed in young floral meristems and in yeast interacts preferentially with the TGA transcription factor encoded by PERIANTHIA (PAN). In support of a biological relevance for this interaction, we show that bop1 bop2 and pan mutants share a pentamerous arrangement of first whorl floral organs, a patterning defect that is retained in bop1 bop2 pan triple mutants. Our data provide evidence that BOP proteins control patterning via direct interactions with TGA transcription factors and demonstrate that a signaling mechanism similar to that formally associated with plant defense is likely used for the control of developmental patterning. PMID- 15805485 TI - A two-edged role for the transposable element Kiddo in the rice ubiquitin2 promoter. AB - Miniature inverted repeat transposable elements (MITEs) are thought to be a driving force for genome evolution. Although numerous MITEs are found associated with genes, little is known about their function in gene regulation. Whereas the rice ubiquitin2 (rubq2) promoter in rice (Oryza sativa) line IR24 contains two nested MITEs (Kiddo and MDM1), that in line T309 has lost Kiddo, providing an opportunity to understand the role of MITEs in promoter function. No difference in endogenous rubq2 transcript levels between T309 and IR24 was evident using RT PCR. However, promoter analysis using both transient and stably transformed calli revealed that Kiddo contributed some 20% of the total expression. Bisulfite genomic sequencing of the rubq2 promoters revealed specific DNA methylation at both symmetric and asymmetric cytosine residues on the MITE sequences, possibly induced by low levels of homologous transcripts. When methylation of the MITEs was blocked by 5-azacytidine treatment, a threefold increase in the endogenous rubq2 transcript level was detected in IR24 compared with that in T309. Together with the observed MITE methylation pattern, the detection of low levels of transcripts, but not small RNAs, corresponding to Kiddo and MDM1 suggested that RNA-dependent DNA methylation is induced by MITE transcripts. We conclude that, although Kiddo enhances transcription from the rubq2 promoter, this effect is mitigated by sequence-specific epigenetic modification. PMID- 15805486 TI - The sulfate transporter SST1 is crucial for symbiotic nitrogen fixation in Lotus japonicus root nodules. AB - Symbiotic nitrogen fixation (SNF) by intracellular rhizobia within legume root nodules requires the exchange of nutrients between host plant cells and their resident bacteria. Little is known at the molecular level about plant transporters that mediate such exchanges. Several mutants of the model legume Lotus japonicus have been identified that develop nodules with metabolic defects that cannot fix nitrogen efficiently and exhibit retarded growth under symbiotic conditions. Map-based cloning of defective genes in two such mutants, sst1-1 and sst1-2 (for symbiotic sulfate transporter), revealed two alleles of the same gene. The gene is expressed in a nodule-specific manner and encodes a protein homologous with eukaryotic sulfate transporters. Full-length cDNA of the gene complemented a yeast mutant defective in sulfate transport. Hence, the gene was named Sst1. The sst1-1 and sst1-2 mutants exhibited normal growth and development under nonsymbiotic growth conditions, a result consistent with the nodule specific expression of Sst1. Data from a previous proteomic study indicate that SST1 is located on the symbiosome membrane in Lotus nodules. Together, these results suggest that SST1 transports sulfate from the plant cell cytoplasm to the intracellular rhizobia, where the nutrient is essential for protein and cofactor synthesis, including nitrogenase biosynthesis. This work shows the importance of plant sulfate transport in SNF and the specialization of a eukaryotic transporter gene for this purpose. PMID- 15805487 TI - N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. AB - Cryptochromes (CRY) are blue light receptors that share sequence similarity with photolyases, flavoproteins that catalyze the repair of UV light-damaged DNA. Transgenic Arabidopsis thaliana seedlings expressing the C-terminal domains of the Arabidopsis CRY fused to beta-glucuronidase (GUS) display a constitutive photomorphogenic (COP) phenotype, indicating that the signaling mechanism of Arabidopsis CRY is mediated through the C-terminal domain. The role of the Arabidopsis CRY N-terminal photolyase-like domain in CRY action remains poorly understood. Here, we report the essential role of the Arabidopsis CRY1 N-terminal domain (CNT1) in the light activation of CRY1 photoreceptor activity. Yeast two hybrid assay, in vitro binding, in vivo chemical cross-linking, gel filtration, and coimmunoprecipitation studies indicate that CRY1 homodimerizes in a light independent manner. Mutagenesis and transgenic studies demonstrate that CNT1 mediated dimerization is required for light activation of the C-terminal domain of CRY1 (CCT1). Transgenic data and native gel electrophoresis studies suggest that multimerization of GUS is both responsible and required for mediating a COP phenotype on fusion to CCT1. These results indicate that the properties of the GUS multimer are analogous to those of the light-modified CNT1 dimer. Irradiation with blue light modifies the properties of the CNT1 dimer, resulting in a change in CCT1, activating CCT1, and eventually triggering the CRY1 signaling pathway. PMID- 15805488 TI - ODORANT1 regulates fragrance biosynthesis in petunia flowers. AB - Floral scent is important to plant reproduction because it attracts pollinators to the sexual organs. Therefore, volatile emission is usually tuned to the foraging activity of the pollinators. In Petunia hybrida, volatile benzenoids determine the floral aroma. Although the pathways for benzenoid biosynthesis have been characterized, the enzymes involved are less well understood. How production and emission are regulated is unknown. By targeted transcriptome analyses, we identified ODORANT1 (ODO1), a member of the R2R3-type MYB family, as a candidate for the regulation of volatile benzenoids in Petunia hybrida cv W115 (Mitchell) flowers. These flowers are only fragrant in the evening and at night. Transcript levels of ODO1 increased before the onset of volatile emission and decreased when volatile emission declined. Downregulation of ODO1 in transgenic P. hybrida Mitchell plants strongly reduced volatile benzenoid levels through decreased synthesis of precursors from the shikimate pathway. The transcript levels of several genes in this pathway were reduced by suppression of ODO1 expression. Moreover, ODO1 could activate the promoter of the 5-enol-pyruvylshikimate-3 phosphate synthase gene. Flower pigmentation, which is furnished from the same shikimate precursors, was not influenced because color and scent biosynthesis occur at different developmental stages. Our studies identify ODO1 as a key regulator of floral scent biosynthesis. PMID- 15805489 TI - Dynamics of COPII vesicles and the Golgi apparatus in cultured Nicotiana tabacum BY-2 cells provides evidence for transient association of Golgi stacks with endoplasmic reticulum exit sites. AB - Despite the ubiquitous presence of the COPI, COPII, and clathrin vesicle budding machineries in all eukaryotes, the organization of the secretory pathway in plants differs significantly from that in yeast and mammalian cells. Mobile Golgi stacks and the lack of both transitional endoplasmic reticulum (ER) and a distinct ER-to-Golgi intermediate compartment are the most prominent distinguishing morphological features of the early secretory pathway in plants. Although the formation of COPI vesicles at periphery of Golgi cisternae has been demonstrated in plants, exit from the ER has been difficult to visualize, and the spatial relationship of this event is now a matter of controversy. Using tobacco (Nicotiana tabacum) BY-2 cells, which represent a highly active secretory system, we have used two approaches to investigate the location and dynamics of COPII binding to the ER and the relationship of these ER exit sites (ERES) to the Golgi apparatus. On the one hand, we have identified endogenous COPII using affinity purified antisera generated against selected COPII-coat proteins (Sar1, Sec13, and Sec23); on the other hand, we have prepared a BY-2 cell line expressing Sec13:green fluorescent protein (GFP) to perform live cell imaging with red fluorescent protein-labeled ER or Golgi stacks. COPII binding to the ER in BY-2 cells is visualized as fluorescent punctate structures uniformly distributed over the surface of the ER, both after antibody staining as well as by Sec13:GFP expression. These structures are smaller and greatly outnumber the Golgi stacks. They are stationary, but have an extremely short half-life (<10 s). Without correlative imaging data on the export of membrane or lumenal ER cargo it was not possible to equate unequivocally these COPII binding loci with ERES. When a GDP fixed Sar1 mutant is expressed, ER export is blocked and the visualization of COPII binding is perturbed. On the other hand, when secretion is inhibited by brefeldin A, COPII binding sites on the ER remain visible even after the Golgi apparatus has been lost. Live cell imaging in a confocal laser scanning microscope equipped with spinning disk optics allowed us to investigate the relationship between mobile Golgi stacks and COPII binding sites. As they move, Golgi stacks temporarily associated with COPII binding sites at their rims. Golgi stacks were visualized with their peripheries partially or fully occupied with COPII. In the latter case, Golgi stacks had the appearance of a COPII halo. Slow moving Golgi stacks tended to have more peripheral COPII than faster moving ones. However, some stationary Golgi stacks entirely lacking COPII were also observed. Our results indicate that, in a cell type with highly mobile Golgi stacks like tobacco BY-2, the Golgi apparatus is not continually linked to a single ERES. By contrast, Golgi stacks associate intermittently and sometimes concurrently with several ERES as they move. PMID- 15805490 TI - Whole genome shotgun sequencing of Brassica oleracea and its application to gene discovery and annotation in Arabidopsis. AB - Through comparative studies of the model organism Arabidopsis thaliana and its close relative Brassica oleracea, we have identified conserved regions that represent potentially functional sequences overlooked by previous Arabidopsis genome annotation methods. A total of 454,274 whole genome shotgun sequences covering 283 Mb (0.44 x) of the estimated 650 Mb Brassica genome were searched against the Arabidopsis genome, and conserved Arabidopsis genome sequences (CAGSs) were identified. Of these 229,735 conserved regions, 167,357 fell within or intersected existing gene models, while 60,378 were located in previously unannotated regions. After removal of sequences matching known proteins, CAGSs that were close to one another were chained together as potentially comprising portions of the same functional unit. This resulted in 27,347 chains of which 15,686 were sufficiently distant from existing gene annotations to be considered a novel conserved unit. Of 192 conserved regions examined, 58 were found to be expressed in our cDNA populations. Rapid amplification of cDNA ends (RACE) was used to obtain potentially full-length transcripts from these 58 regions. The resulting sequences led to the creation of 21 gene models at 17 new Arabidopsis loci and the addition of splice variants or updates to another 19 gene structures. In addition, CAGSs overlapping already annotated genes in Arabidopsis can provide guidance for manual improvement of existing gene models. Published genome-wide expression data based on whole genome tiling arrays and massively parallel signature sequencing were overlaid on the Brassica-Arabidopsis conserved sequences, and 1399 regions of intersection were identified. Collectively our results and these data sets suggest that several thousand new Arabidopsis genes remain to be identified and annotated. PMID- 15805491 TI - Comparing low coverage random shotgun sequence data from Brassica oleracea and Oryza sativa genome sequence for their ability to add to the annotation of Arabidopsis thaliana. AB - Since the completion of the Arabidopsis thaliana genome sequence, there is an ongoing effort to annotate the genome as accurately as possible. Comparing genome sequences of related species complements the current annotation strategies by identifying genes and improving gene structure. A total of 595,321 Brassica oleracea shotgun reads were sequenced by TIGR (The Institute for Genome Research) and the collaboration of Washington University and Cold Spring Harbor. Vicogenta (a genome viewer based on GMOD and GBrowse) was created to view the current annotation and sequence alignments for Arabidopsis. Brassica reads were compared with the Arabidopsis genome and proteome databases using BLAST. Hypothetical genes and conserved unannotated regions on the short arm of chromosome 4 from Arabidopsis were experimentally verified using RT-PCR. We were able to improve the Arabidopsis annotation by identifying 25 genes that were missed, and confirming expression of 43 hypothetical genes in Arabidopsis. We were also able to detect conservation in genes whose transcription is normally suppressed due to methylation. We also examined how useful the O. sativa genome and ESTs from other species are, compared with Brassica, in improving the Arabidopsis annotation. PMID- 15805492 TI - Comparative genome analyses of Arabidopsis spp.: inferring chromosomal rearrangement events in the evolutionary history of A. thaliana. AB - Comparative genome analysis is a powerful tool that can facilitate the reconstruction of the evolutionary history of the genomes of modern-day species. The model plant Arabidopsis thaliana with its n = 5 genome is thought to be derived from an ancestral n = 8 genome. Pairwise comparative genome analyses of A. thaliana with polyploid and diploid Brassicaceae species have suggested that rapid genome evolution, manifested by chromosomal rearrangements and duplications, characterizes the polyploid, but not the diploid, lineages of this family. In this study, we constructed a low-density genetic linkage map of Arabidopsis lyrata ssp. lyrata (A. l. lyrata; n = 8, diploid), the closest known relative of A. thaliana (MRCA approximately 5 Mya), using A. thaliana-specific markers that resolve into the expected eight linkage groups. We then performed comparative Bayesian analyses using raw mapping data from this study and from a Capsella study to infer the number and nature of rearrangements that distinguish the n = 8 genomes of A. l. lyrata and Capsella from the n = 5 genome of A. thaliana. We conclude that there is strong statistical support in favor of the parsimony scenarios of 10 major chromosomal rearrangements separating these n = 8 genomes from A. thaliana. These chromosomal rearrangement events contribute to a rate of chromosomal evolution higher than previously reported in this lineage. We infer that at least seven of these events, common to both sets of data, are responsible for the change in karyotype and underlie genome reduction in A. thaliana. PMID- 15805493 TI - Ancient haplotypes resulting from extensive molecular rearrangements in the wheat A genome have been maintained in species of three different ploidy levels. AB - Plant genomes, in particular grass genomes, evolve very rapidly. The closely related A genomes of diploid, tetraploid, and hexaploid wheat are derived from a common ancestor that lived <3 million years ago and represent a good model to study molecular mechanisms involved in such rapid evolution. We have sequenced and compared physical contigs at the Lr10 locus on chromosome 1AS from diploid (211 kb), tetraploid (187 kb), and hexaploid wheat (154 kb). A maximum of 33% of the sequences were conserved between two species. The sequences from diploid and tetraploid wheat shared all of the genes, including Lr10 and RGA2 and define a first haplotype (H1). The 130-kb intergenic region between Lr10 and RGA2 was conserved in size despite its activity as a hot spot for transposon insertion, which resulted in >70% of sequence divergence. The hexaploid wheat sequence lacks both Lr10 and RGA2 genes and defines a second haplotype, H2, which originated from ancient and extensive rearrangements. These rearrangements included insertions of retroelements and transposons deletions, as well as unequal recombination within elements. Gene disruption in haplotype H2 was caused by a deletion and subsequent large inversion. Gene conservation between H1 haplotypes, as well as conservation of rearrangements at the origin of the H2 haplotype at three different ploidy levels indicate that the two haplotypes are ancient and had a stable gene content during evolution, whereas the intergenic regions evolved rapidly. Polyploidization during wheat evolution had no detectable consequences on the structure and evolution of the two haplotypes. PMID- 15805494 TI - Functional insights from the distribution and role of homopeptide repeat containing proteins. AB - Expansion of "low complex" repeats of amino acids such as glutamine (Poly-Q) is associated with protein misfolding and the development of degenerative diseases such as Huntington's disease. The mechanism by which such regions promote misfolding remains controversial, the function of many repeat-containing proteins (RCPs) remains obscure, and the role (if any) of repeat regions remains to be determined. Here, a Web-accessible database of RCPs is presented. The distribution and evolution of RCPs that contain homopeptide repeats tracts are considered, and the existence of functional patterns investigated. Generally, it is found that while polyamino acid repeats are extremely rare in prokaryotes, several eukaryote putative homologs of prokaryote RCP-involved in important housekeeping processes-retain the repetitive region, suggesting an ancient origin for certain repeats. Within eukarya, the most common uninterrupted amino acid repeats are glutamine, asparagines, and alanine. Interestingly, while poly-Q repeats are found in vertebrates and nonvertebrates, poly-N repeats are only common in more primitive nonvertebrate organisms, such as insects and nematodes. We have assigned function to eukaryote RCPs using Online Mendelian Inheritance in Man (OMIM), the Human Reference Protein Database (HRPD), FlyBase, and Wormpep. Prokaryote RCPs were annotated using BLASTp searches and Gene Ontology. These data reveal that the majority of RCPs are involved in processes that require the assembly of large, multiprotein complexes, such as transcription and signaling. PMID- 15805495 TI - Novel specificities emerge by stepwise duplication of functional modules. AB - A functional module can be defined as a spatially or chemically isolated set of functionally associated components that accomplishes a discrete biological process. Modularity is a key attribute of cellular systems, but the mechanisms that underlie the evolution of functional modules are largely unknown. Duplication of modules has been shown to be an efficient mechanism for the generation of functional innovation in the field of artificial intelligence, but has not been studied in biological networks. Therefore, we ask whether module duplication occurs in cellular networks. We developed a generic framework for the analysis of module duplication, and use it in a large-scale analysis of Saccharomyces cerevisiae protein complexes. Protein complexes are well defined, experimentally derived, functional modules. We observe that at least 6%-20% of the protein complexes have strong similarity to other complexes; thus a considerable fraction has evolved by duplication. Our results indicate that many complexes evolved by step-wise partial duplications. We show that duplicated complexes retain the same overall function, but have different binding specificities and regulation, revealing that duplication of these modules is associated with functional specialization. PMID- 15805496 TI - Forcing interactions as a genetic screen to identify proteins that exert a defined activity. AB - The interaction of proteins to form macromolecular complexes is the basis for most biological processes. Approaches have been described that employ artificial constructs to promote such complexes and assess the consequences. For example, a protein interaction scheme has been described that examines the effects of a specific phosphorylation event catalyzed by a protein kinase via the provision of an artificial protein binding interface between a modified version of the kinase and a single substrate. We have generalized this type of approach to form the basis for a genetic selection to identify proteins that exert an activity when recruited to a target protein. The assay uses the leucine zipper domains from the mammalian transcription factors Fos and Jun to force the interaction of two proteins. With a target protein fused to the Jun zipper and a library of open reading frames fused to the Fos zipper, we demonstrate this approach in yeast with both a selection to identify membrane-associated proteins and a selection to identify candidate components of the filamentous growth MAP kinase pathway. PMID- 15805497 TI - ECgene: genome-based EST clustering and gene modeling for alternative splicing. AB - With the availability of the human genome map and fast algorithms for sequence alignment, genome-based EST clustering became a viable method for gene modeling. We developed a novel gene-modeling method, ECgene (Gene modeling by EST Clustering), which combines genome-based EST clustering and the transcript assembly procedure in a coherent and consistent fashion. Specifically, ECgene takes alternative splicing events into consideration. The position of splice sites (i.e., exon-intron boundaries) in the genome map is utilized as the critical information in the whole procedure. Sequences that share any splice sites are grouped together to define an EST cluster in a manner similar to that of the genome-based version of the UniGene algorithm. Transcript assembly is achieved using graph theory that represents the exon connectivity in each cluster as a directed acyclic graph (DAG). Distinct paths along exons correspond to possible gene models encompassing all alternative splicing events. EST sequences in each cluster are subclustered further according to the compatibility with gene structure of each splice variant, and they can be regarded as clone evidence for the corresponding isoform. The reliability of each isoform is assessed from the nature of cluster members and from the minimum number of clones required to reconstruct all exons in the transcript. PMID- 15805498 TI - Closing in on the C. elegans ORFeome by cloning TWINSCAN predictions. AB - The genome of Caenorhabditis elegans was the first animal genome to be sequenced. Although considerable effort has been devoted to annotating it, the standard WormBase annotation contains thousands of predicted genes for which there is no cDNA or EST evidence. We hypothesized that a more complete experimental annotation could be obtained by creating a more accurate gene-prediction program and then amplifying and sequencing predicted genes. Our approach was to adapt the TWINSCAN gene prediction system to C. elegans and C. briggsae and to improve its splice site and intron-length models. The resulting system has 60% sensitivity and 58% specificity in exact prediction of open reading frames (ORFs), and hence, proteins-the best results we are aware of any multicellular organism. We then attempted to amplify, clone, and sequence 265 TWINSCAN-predicted ORFs that did not overlap WormBase gene annotations. The success rate was 55%, adding 146 genes that were completely absent from WormBase to the ORF clone collection (ORFeome). The same procedure had a 7% success rate on 90 Worm Base "predicted" genes that do not overlap TWINSCAN predictions. These results indicate that the accuracy of WormBase could be significantly increased by replacing its partially curated predicted genes with TWINSCAN predictions. The technology described in this study will continue to drive the C. elegans ORFeome toward completion and contribute to the annotation of the three Caenorhabditis species currently being sequenced. The results also suggest that this technology can significantly improve our knowledge of the "parts list" for even the best-studied model organisms. PMID- 15805499 TI - Construction of a mini-Tn5-luxCDABE mutant library in Pseudomonas aeruginosa PAO1: a tool for identifying differentially regulated genes. AB - Pseudomonas aeruginosa is a major cause of nosocomial (hospital-derived) infections, is the predominant pathogen in chronic cystic fibrosis lung infections, and remains difficult to treat due to its high intrinsic antibiotic resistance. The completion of the P. aeruginosa PAO1 genome sequence provides the opportunity for genome-wide studies to increase our understanding of the pathogenesis and biology of this important pathogen. In this report, we describe the construction of a mini-Tn5-luxCDABE mutant library and a high-throughput inverse PCR method to amplify DNA flanking the site of insertion for sequencing and insertion site mapping. In addition to producing polar knockout mutations in nonessential genes, the promoterless luxCDABE reporter present in the transposon serves as a real-time reporter of gene expression for the inactivated gene. A total of 2519 transposon insertion sites were mapped, 77% of which were nonredundant insertions. Of the insertions within an ORF, -55% of total and unique insertion sites were transcriptional luxCDABE fusions. A bias toward low insertion-site density in the genome region that surrounds the predicted terminus of replication was observed. To demonstrate the utility of chromosomal lux fusions, we performed extensive regulatory screens to identify genes that were differentially regulated under magnesium or phosphate limitation. This approach led to the discovery of many known and novel genes necessary for these environmental adaptations, including genes involved in resistance to cationic antimicrobial peptides. This dual-purpose mutant library allows for functional and regulation studies and will serve as a resource for the research community to further our understanding of P. aeruginosa biology. PMID- 15805500 TI - CADLIVE dynamic simulator: direct link of biochemical networks to dynamic models. AB - We have developed the CADLIVE (Computer-Aided Design of LIVing systEms) Simulator that provided a rule-based automatic way to convert biochemical network maps into dynamic models, which enables simulating their dynamics without going through all of the reactions down to the details of exact kinetic parameters. The simulator supports the biochemical reaction maps that are generated by the previously developed GUI editor. Notice that the part of the GUI editor had been previously published, but, as yet, not the simulator. To directly link biochemical network maps to dynamic simulation, we have created the strategy of three layers and two stages with the efficient conversion rules in an XML representation. This strategy divides a molecular network into three layers, i.e., gene, protein, and metabolic layers, and partitions the conversion process into two stages. Once a biochemical map is provided, CADLIVE automatically builds a mathematical model, thereby facilitating one to simulate and analyze it. In order to demonstrate the feasibility of CADLIVE, we analyzed the Escherichia coli nitrogen-assimilation system (64 equations with 64 variables) that consists of multiple and complicated negative and positive feedback loops. CADLIVE predicted that the glnK gene is responsible for hysteresis or reversibility of nitrogen-related (Ntr) gene expression with respect to the ammonia concentration, supporting the experimental observation of the runaway expression of the Ntr genes. PMID- 15805501 TI - A novel mutation in kaiC affects resetting of the cyanobacterial circadian clock. AB - Light is the most important factor controlling circadian systems in response to day-night cycles. In order to better understand the regulation of circadian rhythms by light in Synechococcus elongatus PCC 7942, we screened for mutants with defective phase shifting in response to dark pulses. Using a 5-h dark-pulse protocol, we identified a mutation in kaiC that we termed pr1, for phase response 1. In the pr1 mutant, a 5-h dark pulse failed to shift the phase of the circadian rhythm, while the same pulse caused a 10-h phase shift in wild-type cells. The rhythm in accumulation of KaiC was abolished in the pr1 mutant, and the rhythmicity of KaiC phosphorylation was reduced. Additionally, the pr1 mutant was defective in mediating the feedback inhibition of kaiBC. Finally, overexpression of mutant KaiC led to a reduced phase shift compared to that for wild-type KaiC. Thus, KaiC appears to play a role in resetting the cellular clock in addition to its documented role in the feedback regulation of circadian rhythms. PMID- 15805502 TI - Genome-wide transcriptional profiling of the steady-state response of Pseudomonas aeruginosa to hydrogen peroxide. AB - The steady-state response of Pseudomonas aeruginosa to the oxidative-stress generating agent hydrogen peroxide was analyzed by PAO1 transcriptome profiling. In total, 694, 411, and 237 genes were upregulated and 668, 576, and 468 genes were downregulated in P. aeruginosa strains TB, 892, and PAO1, respectively. The expression profiles of the two variants of the TB clone were significantly more related to each other than the expression profile of either strain was to that of PAO1. Exposure to H(2)O(2) activated by more than 10-fold the expression of the cyoABCD operon, which is key for aerobic respiration, and of oxidative-stress response elements such as the catalase KatB, the alkyl hydroperoxide reductase AhpF, and the thioredoxin reductase 2 operon. Genes for iron and sulfur homeostasis were upregulated. Most enzymes necessary for the conversion of amino acids into the citric acid cycle were globally downregulated at the transcriptional level. Nitrate respiration and arginine fermentation were shut off in the clone TB strains and attenuated in the PAO strain. The transcriptional profiles indicate that the two clone TB strains are more proficient in coping with H(2)O(2)-mediated oxidative stress than the reference strain PAO. According to this data, we recommend study of the transcriptome of strain PAO1 in parallel with those of at least two strains of another clone in order to differentiate common responses from clone- and strain-specific responses and to minimize overinterpretations of microarray data. PMID- 15805503 TI - Dual roles of an E-helix residue, Glu167, in the transcriptional activator function of CooA. AB - CooA is a transcriptional activator that mediates CO-dependent expression of the genes responsible for CO oxidation in Rhodospirillum rubrum. In this study, we suggest in vitro and in vivo models explaining an unusual requirement of CooA for millimolar levels of divalent cations for high-affinity DNA binding. Several lines of evidence indicate that an E-helix residue, Glu167, plays a central role in this requirement by inhibiting sequence-specific DNA binding via charge repulsion in the absence of any divalent cation and that divalent cations relieve such repulsion in the process of DNA binding by CooA. Unexpectedly, the Glu167 residue is the optimal residue for in vivo transcriptional activity of CooA. We present a model in which the Glu167 from the downstream subunit of CooA helps the protein to interact with RNA polymerase, probably through an interaction between activating region 3 and sigma subunit. The study was further extended to a homologous protein, cyclic AMP receptor protein (CRP), which revealed similar, but not identical, roles of the residue in this protein as well. The results show a unique mechanism of CooA modulating its DNA binding and transcriptional activation in response to divalent cations among the CRP/FNR (fumarate and nitrate reductase activator protein) superfamily of regulators. PMID- 15805504 TI - Characterization of LtsA from Rhodococcus erythropolis, an enzyme with glutamine amidotransferase activity. AB - The nocardioform actinomycete Rhodococcus erythropolis has a characteristic cell wall structure. The cell wall is composed of arabinogalactan and mycolic acid and is highly resistant to the cell wall-lytic activity of lysozyme (muramidase). In order to improve the isolation of recombinant proteins from R. erythropolis host cells (N. Nakashima and T. Tamura, Biotechnol. Bioeng. 86:136-148, 2004), we isolated two mutants, L-65 and L-88, which are susceptible to lysozyme treatment. The lysozyme sensitivity of the mutants was complemented by expression of Corynebacterium glutamicum ltsA, which codes for an enzyme with glutamine amidotransferase activity that results from coupling of two reactions (a glutaminase activity and a synthetase activity). The lysozyme sensitivity of the mutants was also complemented by ltsA homologues from Bacillus subtilis and Mycobacterium tuberculosis, but the homologues from Streptomyces coelicolor and Escherichia coli did not complement the sensitivity. This result suggests that only certain LtsA homologues can confer lysozyme resistance. Wild-type recombinant LtsA from R. erythropolis showed glutaminase activity, but the LtsA enzymes from the L-88 and L-65 mutants displayed drastically reduced activity. Interestingly, an ltsA disruptant mutant, which expressed the mutated LtsA, changed from lysozyme sensitive to lysozyme resistant when NH(4)Cl was added into the culture media. The glutaminase activity of the LtsA mutants inactivated by site-directed mutagenesis was also restored by addition of NH(4)Cl, indicating that NH(3) can be used as an amide donor molecule. Taken together, these results suggest that LtsA is critically involved in mediating lysozyme resistance in R. erythropolis cells. PMID- 15805505 TI - Expression of the bmpB gene of Borrelia burgdorferi is modulated by two distinct transcription termination events. AB - bmp gene family 36 of Borrelia burgdorferi, the agent of Lyme disease, comprises four paralogs: bmpA, bmpB, bmpC, and bmpD. The bmpA and bmpB genes constitute an operon. All four genes have been found to be transcribed in cultured spirochetes. Expression from the bmpAB operon results in three distinct transcripts of 1.1, 1.6, and 2.4 kb, and the relative expression of bmpA mRNA is three- to fourfold greater than that of bmpB mRNA. However, thus far only expression of the BmpA protein has been demonstrated. Therefore, in this study we characterized the origins of the three transcripts and compared the relative expression of the BmpA and BmpB proteins. Northern blotting revealed that the three distinct transcripts originated from a single promoter located upstream of bmpA but terminated either 3' to the bmpA (1.1-kb RNA) or bmpB (2.4-kb RNA) gene or, most unusually, within the bmpB gene (1.6-kb RNA). Termination within the bmpB gene was associated with a functional Rho-independent transcription terminator. At the protein level, we also observed a 4.3-fold greater abundance of BmpA compared to that of BmpB. These studies identify a transcription termination mechanism in B. burgdorferi resulting in the disparate expression of the two genes of the bmpAB operon. PMID- 15805506 TI - The metalloprotease of Listeria monocytogenes controls cell wall translocation of the broad-range phospholipase C. AB - Listeria monocytogenes is a gram-positive bacterial pathogen that multiplies in the cytosol of host cells and spreads directly from cell to cell. During cell-to cell spread, bacteria become temporarily confined to secondary vacuoles. The broad-range phospholipase C (PC-PLC) of L. monocytogenes contributes to bacterial escape from secondary vacuoles. PC-PLC requires cleavage of an N-terminal propeptide for activation, and Mpl, a metalloprotease of Listeria, is involved in the proteolytic activation of PC-PLC. Previously, we showed that cell wall translocation of PC-PLC is inefficient, resulting in accumulation of PC-PLC at the membrane-cell wall interface. In infected cells, rapid cell wall translocation of PC-PLC is triggered by a decrease in pH and correlates with cleavage of the propeptide in an Mpl-dependent manner. To address the role of the propeptide and of Mpl in cell wall translocation of PC-PLC, we generated a cleavage site mutant and a propeptide deletion mutant. The intracellular behavior of these mutants was assessed in pulse-chase experiments. We observed efficient translocation of the proform of the PC-PLC cleavage site mutant in a manner that was pH sensitive and Mpl dependent. However, the propeptide deletion mutant was efficiently translocated into host cells independent of Mpl and pH. Overall, these results suggest that Mpl regulates PC-PLC translocation across the bacterial cell wall in a manner that is dependent on the presence of the propeptide but independent of propeptide cleavage. In addition, similarly to Mpl mediated cleavage of PC-PLC propeptide, Mpl-mediated translocation of PC-PLC across the bacterial cell wall is pH sensitive. PMID- 15805507 TI - Expression of fnr is constrained by an upstream IS5 insertion in certain Escherichia coli K-12 strains. AB - FNR is a global transcriptional regulator that controls anaerobic gene expression in Escherichia coli. Through the use of a number of approaches it was shown that fnr gene expression is reduced approximately three- to fourfold in E. coli strain MC4100 compared with the results seen with strain MG1655. This reduction in fnr expression is due to the insertion of IS5 (is5F) in the regulatory region of the gene at position -41 relative to the transcription initiation site. Transcription of the fnr gene nevertheless occurs from its own promoter in strain MC4100, but transcript levels are reduced approximately fourfold compared with those seen with strain MG1655. Remarkably, in strains bearing is5F the presence of Hfq prevents IS5-dependent transcriptional silencing of fnr expression. Thus, an hfq mutant of MC4100 is devoid of FNR protein and has the phenotype of an fnr mutant. In strain MG1655, or a derivative of MC4100 lacking is5F, mutation of hfq had no effect on fnr transcript levels. This finding indicates that IS5 mediates the effect of Hfq on fnr expression in MC4100. Western blot analysis revealed that cellular levels of FNR were reduced threefold in strain MC4100 compared with strain MG1655 results. A selection of FNR-dependent genes fused to lacZ were analyzed for the effects of reduced FNR levels on anaerobic gene expression. Expression of some operons, e.g., focA-pfl and fdnGHJI, was unaffected by reduction in the level of FNR, while the expression of other genes such as ndh and nikA was clearly affected. PMID- 15805508 TI - An FNR-type regulator controls the anaerobic expression of hyn hydrogenase in Thiocapsa roseopersicina. AB - The purple sulfur photosynthetic bacterium Thiocapsa roseopersicina BBS contains a heat-stable membrane-associated hydrogenase encoded by the hyn operon. Expression from the hyn operon regulatory region is up-regulated under anaerobic conditions. cis elements were mapped between positions -602 and -514 upstream from the hynS gene. Within this region two sequences that resemble DNA sites for FNR were recognized. The gene of an FNR homologue, FnrT, was identified in the genome of T. roseopersicina, and an fnrT knockout mutant was constructed. Anaerobic induction of hynS expression was abolished in the fnrT mutant, suggesting that FnrT is an activator of the hynS promoter. The T. roseopersicina hynS promoter could be activated in Escherichia coli, and this regulation was dependent on E. coli FNR. In vitro experiments with purified E. coli Ala154 FNR protein and purified E. coli RNA polymerase showed that FNR bound to two sites in the hyn regulatory region, that FNR could activate transcription initiation at the hynS promoter, and that FNR bound at the two target sites activated to different extents. PMID- 15805509 TI - Flavobacterium johnsoniae GldJ is a lipoprotein that is required for gliding motility. AB - Cells of Flavobacterium johnsoniae glide rapidly over surfaces by an unknown mechanism. Eight genes required for gliding motility have been described. Complementation of the nonmotile mutant UW102-48 identified another gene, gldJ, that is required for gliding. gldJ mutants formed nonspreading colonies, and individual cells were completely nonmotile. Like previously described nonmotile mutants, gldJ mutants were deficient in chitin utilization and were resistant to bacteriophages that infect wild-type cells. Cell fractionation and labeling studies with [(3)H]palmitate indicated that GldJ is a lipoprotein. Mutations in gldA, gldB, gldD, gldF, gldG, gldH, or gldI resulted in normal levels of gldJ transcript but decreased levels of GldJ protein. Expression of truncated GldJ protein in wild-type cells resulted in a severe motility defect. GldJ was found in regular bands that suggest the presence of a helical structure within the cell envelope. PMID- 15805510 TI - Diversity of genome structure in Salmonella enterica serovar Typhi populations. AB - The genomes of most strains of Salmonella and Escherichia coli are highly conserved. In contrast, all 136 wild-type strains of Salmonella enterica serovar Typhi analyzed by partial digestion with I-CeuI (an endonuclease which cuts within the rrn operons) and pulsed-field gel electrophoresis and by PCR have rearrangements due to homologous recombination between the rrn operons leading to inversions and translocations. Recombination between rrn operons in culture is known to be equally frequent in S. enterica serovar Typhi and S. enterica serovar Typhimurium; thus, the recombinants in S. enterica serovar Typhi, but not those in S. enterica serovar Typhimurium, are able to survive in nature. However, even in S. enterica serovar Typhi the need for genome balance and the need for gene dosage impose limits on rearrangements. Of 100 strains of genome types 1 to 6, 72 were only 25.5 kb off genome balance (the relative lengths of the replichores during bidirectional replication from oriC to the termination of replication [Ter]), while 28 strains were less balanced (41 kb off balance), indicating that the survival of the best-balanced strains was greater. In addition, the need for appropriate gene dosage apparently selected against rearrangements which moved genes from their accustomed distance from oriC. Although rearrangements involving the seven rrn operons are very common in S. enterica serovar Typhi, other duplicated regions, such as the 25 IS200 elements, are very rarely involved in rearrangements. Large deletions and insertions in the genome are uncommon, except for deletions of Salmonella pathogenicity island 7 (usually 134 kb) from fragment I-CeuI-G and 40-kb insertions, possibly a prophage, in fragment I-CeuI-E. The phage types were determined, and the origins of the phage types appeared to be independent of the origins of the genome types. PMID- 15805511 TI - Protein diversity confers specificity in plasmid segregation. AB - The ParG segregation protein (8.6 kDa) of multidrug resistance plasmid TP228 is a homodimeric DNA-binding factor. The ParG dimer consists of intertwined C-terminal domains that adopt a ribbon-helix-helix architecture and a pair of flexible, unstructured N-terminal tails. A variety of plasmids possess partition loci with similar organizations to that of TP228, but instead of ParG homologs, these plasmids specify a diversity of unrelated, but similarly sized, partition proteins. These include the proteobacterial pTAR, pVT745, and pB171 plasmids. The ParG analogs of these plasmids were characterized in parallel with the ParG homolog encoded by the pseudomonal plasmid pVS1. Like ParG, the four proteins are dimeric. No heterodimerization was detectable in vivo among the proteins nor with the prototypical ParG protein, suggesting that monomer-monomer interactions are specific among the five proteins. Nevertheless, as with ParG, the ParG analogs all possess significant amounts of unordered amino acid residues, potentially highlighting a common structural link among the proteins. Furthermore, the ParG analogs bind specifically to the DNA regions located upstream of their homologous parF-like genes. These nucleoprotein interactions are largely restricted to cognate protein-DNA pairs. The results reveal that the partition complexes of these and related plasmids have recruited disparate DNA-binding factors that provide a layer of specificity to the macromolecular interactions that mediate plasmid segregation. PMID- 15805512 TI - The Bacillus subtilis SinR and RapA developmental regulators are responsible for inhibition of spore development by alcohol. AB - Even though there is a large body of information concerning the harmful effects of alcohol on different organisms, the mechanism(s) that affects developmental programs, at a single-cell level, has not been clearly identified. In this respect, the spore-forming bacterium Bacillus subtilis constitutes an excellent model to study universal questions of cell fate, cell differentiation, and morphogenesis. Here, we demonstrate that treatment with subinhibitory concentrations of alcohol that did not affect vegetative growth inhibited the initiation of spore development through a selective blockage of key developmental genes under the control of the master transcription factor Spo0A approximately P. Isopropyl-beta-D-thiogalactopyranoside-directed expression of a phosphorylation independent form of Spo0A (Sad67) and the use of an in vivo mini-Tn10 insertional library permitted the identification of the developmental SinR repressor and RapA phosphatase as the effectors that mediated the inhibitory effect of alcohol on spore morphogenesis. A double rapA sinR mutant strain was completely resistant to the inhibitory effects of different-C-length alcohols on sporulation, indicating that the two cell fate determinants were the main or unique regulators responsible for the spo0 phenotype of wild-type cells in the presence of alcohol. Furthermore, treatment with alcohol produced a significant induction of rapA and sinR, while the stationary-phase induction of sinI, which codes for a SinR inhibitor, was completely turned off by alcohol. As a result, a dramatic repression of spo0A and the genes under its control occurred soon after alcohol addition, inhibiting the onset of sporulation and permitting the evaluation of alternative pathways required for cellular survival. PMID- 15805513 TI - Translational control of tetracycline resistance and conjugation in the Bacteroides conjugative transposon CTnDOT. AB - The tetQ-rteA-rteB operon of the Bacteroides conjugative transposon CTnDOT is responsible for tetracycline control of the excision and transfer of CTnDOT. Previous studies revealed that tetracycline control of this operon occurred at the translational level and involved a hairpin structure located within the 130 base leader sequence that lies between the promoter of tetQ and the start codon of the gene. This hairpin structure is formed by two sequences, designated Hp1 and Hp8. Hp8 contains the ribosome binding site for tetQ. Examination of the leader region sequence revealed three sequences that might encode a leader peptide. One was only 3 amino acids long. The other two were 16 amino acids long. By introducing stop codons into the peptide coding regions, we have now shown that the 3-amino-acid peptide is the one that is essential for tetracycline control. Between Hp1 and Hp8 lies an 85-bp region that contains other possible RNA hairpin structures. Deletion analysis of this intervening DNA segment has now identified a sequence, designated Hp2, which is essential for tetracycline regulation. This sequence could form a short hairpin structure with Hp1. Mutations that made the Hp1-Hp2 structure more stable caused nearly constitutively high expression of the operon. Thus, stalling of ribosomes on the 3-amino-acid leader peptide could favor formation of the Hp1-Hp2 structure and thus preclude formation of the Hp1-Hp8 structure, releasing the ribosome binding site of tetQ. Finally, comparison of the CTnDOT tetQ leader regions with upstream regions of five tetQ genes found in other elements reveals that the sequences are virtually identical, suggesting that translational attenuation is responsible for control of tetracycline resistance in these other cases as well. PMID- 15805514 TI - Identification of cyclic AMP-regulated genes in Mycobacterium tuberculosis complex bacteria under low-oxygen conditions. AB - Mycobacterium tuberculosis is the etiological agent of tuberculosis (TB), which kills approximately 2 million people a year despite current treatment options. A greater understanding of the biology of this bacterium is needed to better combat TB disease. The M. tuberculosis genome encodes as many as 15 adenylate cyclases, suggesting that cyclic AMP (cAMP) has an important, yet overlooked, role in mycobacteria. This study examined the effect of exogenous cAMP on protein expression in Mycobacterium bovis BCG grown under hypoxic versus ambient conditions. Both shaking and shallow standing cultures were examined for each atmospheric condition. Different cAMP-dependent changes in protein expression were observed in each condition by two-dimensional gel electrophoresis. Shaking low-oxygen cultures produced the most changes (12), while standing ambient conditions showed the fewest (2). Five upregulated proteins, Rv1265, Rv2971, GroEL2, PE_PGRS6a, and malate dehydrogenase, were identified from BCG by mass spectrometry and were shown to also be regulated by cAMP at the mRNA level in both M. tuberculosis H37Rv and BCG. To our knowledge, these data provide the first direct evidence for cAMP-mediated gene regulation in TB complex mycobacteria. PMID- 15805515 TI - Import of the transfer RNase colicin D requires site-specific interaction with the energy-transducing protein TonB. AB - The transfer RNase colicin D and ionophoric colicin B appropriate the outer membrane iron siderophore receptor FepA and share a common translocation requirement for the TonB pathway to cross the outer membrane. Despite the almost identical sequences of the N-terminal domains required for the translocation of colicins D and B, two spontaneous tonB mutations (Arg158Ser and Pro161Leu) completely abolished colicin D toxicity but did not affect either the sensitivity to other colicins or the FepA-dependent siderophore uptake capacity. The sensitivity to colicin D of both tonB mutants was fully restored by specific suppressor mutations in the TonB box of colicin D, at Ser18(Thr) and Met19(Ile), respectively. This demonstrates that the interaction of colicin D with TonB is critically dependent on certain residues close to position 160 in TonB and on the side chains of certain residues in the TonB box of colicin D. The effect of introducing the TonB boxes from other TonB-dependent receptors and colicins into colicins D and B was studied. The results of these and other changes in the two TonB boxes show that the role of residues at positions 18 and 19 in colicin D is strongly modulated by other nearby and/or distant residues and that the overall function of colicin D is much more dependent on the interaction with TonB involving the TonB box than is the function of colicin B. PMID- 15805516 TI - Nucleotide substitution and recombination at orthologous loci in Staphylococcus aureus. AB - The pattern of nucleotide substitution was examined at 2,129 orthologous loci among five genomes of Staphylococcus aureus, which included two sister pairs of closely related genomes (MW2/MSSA476 and Mu50/N315) and the more distantly related MRSA252. A total of 108 loci were unusual in lacking any synonymous differences among the five genomes; most of these were short genes encoding proteins highly conserved at the amino acid sequence level (including many ribosomal proteins) or unknown predicted genes. In contrast, 45 genes were identified that showed anomalously high divergence at synonymous sites. The latter genes were evidently introduced by homologous recombination from distantly related genomes, and in many cases, the pattern of nucleotide substitution made it possible to reconstruct the most probable recombination event involved. These recombination events introduced genes encoding proteins that differed in amino acid sequence and thus potentially in function. Several of the proteins are known or likely to be involved in pathogenesis (e.g., staphylocoagulase, exotoxin, Ser Asp fibrinogen-binding bone sialoprotein-binding protein, fibrinogen and keratin 10 binding surface-anchored protein, fibrinogen-binding protein ClfA, and enterotoxin P). Therefore, the results support the hypothesis that exchange of homologous genes among S. aureus genomes can play a role in the evolution of pathogenesis in this species. PMID- 15805517 TI - Smx nuclease is the major, low-pH-inducible apurinic/apyrimidinic endonuclease in Streptococcus mutans. AB - The causative agent of dental caries in humans, Streptococcus mutans, outcompetes other bacterial species in the oral cavity and causes disease by surviving acidic conditions in dental plaque. We have previously reported that the low-pH survival strategy of S. mutans includes the ability to induce a DNA repair system that appears to involve an enzyme with exonuclease functions (K. Hahn, R. C. Faustoferri, and R. G. Quivey, Jr., Mol. Microbiol 31:1489-1498, 1999). Here, we report overexpression of the S. mutans apurinic/apyrimidinic (AP) endonuclease, Smx, in Escherichia coli; initial characterization of its enzymatic activity; and analysis of an smx mutant strain of S. mutans. Insertional inactivation of the smx gene eliminates the low-pH-inducible exonuclease activity previously reported. In addition, loss of Smx activity renders the mutant strain sensitive to hydrogen peroxide treatment but relatively unaffected by acid-mediated damage or near-UV irradiation. The smx strain of S. mutans was highly sensitive to the combination of iron and hydrogen peroxide, indicating the likely production of hydroxyl radical by Fenton chemistry with concomitant formation of AP sites that are normally processed by the wild-type allele. Smx activity was sufficiently expressed in E. coli to protect an xth mutant strain from the effects of hydrogen peroxide treatment. The data indicate that S. mutans expresses an inducible, class II-like AP endonuclease, encoded by the smx gene, that exhibits exonucleolytic activity and is regulated as part of the acid-adaptive response of the organism. Smx is likely the primary, if not the sole, AP endonuclease induced during growth at low pH values. PMID- 15805518 TI - Completion of the genome sequence of Brucella abortus and comparison to the highly similar genomes of Brucella melitensis and Brucella suis. AB - Brucellosis is a worldwide disease of humans and livestock that is caused by a number of very closely related classical Brucella species in the alpha-2 subdivision of the Proteobacteria. We report the complete genome sequence of Brucella abortus field isolate 9-941 and compare it to those of Brucella suis 1330 and Brucella melitensis 16 M. The genomes of these Brucella species are strikingly similar, with nearly identical genetic content and gene organization. However, a number of insertion-deletion events and several polymorphic regions encoding putative outer membrane proteins were identified among the genomes. Several fragments previously identified as unique to either B. suis or B. melitensis were present in the B. abortus genome. Even though several fragments were shared between only B. abortus and B. suis, B. abortus shared more fragments and had fewer nucleotide polymorphisms with B. melitensis than B. suis. The complete genomic sequence of B. abortus provides an important resource for further investigations into determinants of the pathogenicity and virulence phenotypes of these bacteria. PMID- 15805519 TI - Mutants of FtsZ targeting the protofilament interface: effects on cell division and GTPase activity. AB - The bacterial cell division protein FtsZ assembles into straight protofilaments, one subunit thick, in which subunits appear to be connected by identical bonds or interfaces. These bonds involve the top surface of one subunit making extensive contact with the bottom surface of the subunit above it. We have investigated this interface by site-directed mutagenesis. We found nine bottom and eight top mutants that were unable to function for cell division. We had expected that some of the mutants might poison cell division substoichiometrically, but this was not found for any mutant. Eight of the bottom mutants exhibited dominant negative effects (reduced colony size) and four completely blocked colony formation, but this required expression of the mutant protein at four to five times the wild type FtsZ level. Remarkably, the top mutants were even weaker, most showing no effect at the highest expression level. This suggests a directional assembly or treadmilling, where subunit addition is primarily to the bottom end of the protofilament. Selected pairs of top and bottom mutants showed no GTPase activity up to 10 to 20 microM, in contrast to the high GTPase activity of wild-type FtsZ above 1 muM. Overall, these results suggest that in order for a subunit to bind a protofilament at the 1 microM K(d) for elongation, it must have functional interfaces at both the top and bottom. This is inconsistent with the present model of the protofilament, as a simple stack of subunits one on top of the other, and may require a new structural model. PMID- 15805520 TI - cse, a Chimeric and variable gene, encodes an extracellular protein involved in cellular segregation in Streptococcus thermophilus. AB - The isolation of a Streptococcus thermophilus CNRZ368 mutant displaying a long chain phenotype allowed us to identify the cse gene (for cellular segregation). The N terminus of Cse exhibits high similarity to Streptococcus agalactiae surface immunogenic protein (SIP), while its C terminus exhibits high similarity to S. thermophilus PcsB. In CNRZ368, deletion of the entire cse open reading frame leads to drastic lengthening of cell chains and altered colony morphology. Complementation of the Deltacse mutation with a wild-type allele restored both wild-type phenotypes. The central part of Cse is a repeat-rich region with low sequence complexity. Comparison of cse from CNRZ368 and LMG18311 strains reveals high variability of this repeat-rich region. To assess the impact of this central region variability, the central region of LMG18311 cse was exchanged with that of CNRZ368 cse. This replacement did not affect chain length, showing that divergence of the central part does not modify cell segregation activity of Cse. The structure of the cse locus suggests that the chimeric organization of cse results from insertion of a duplicated sequence deriving from the pcsB 3' end into an ancestral sip gene. Thus, the cse locus illustrates the module-shuffling mechanism of bacterial gene evolution. PMID- 15805521 TI - Mycobacterial lipid II is composed of a complex mixture of modified muramyl and peptide moieties linked to decaprenyl phosphate. AB - Structural analysis of compounds identified as lipid I and II from Mycobacterium smegmatis demonstrated that the lipid moiety is decaprenyl phosphate; thus, M. smegmatis is the first bacterium reported to utilize a prenyl phosphate other than undecaprenyl phosphate as the lipid carrier involved in peptidoglycan synthesis. In addition, mass spectrometry showed that the muropeptides from lipid I are predominantly N-acetylmuramyl-L-alanine-D-glutamate-meso-diaminopimelic acid-D-alanyl-D-alanine, whereas those isolated from lipid II form an unexpectedly complex mixture in which the muramyl residue and the pentapeptide are modified singly and in combination. The muramyl residue is present as N acetylmuramic acid, N-glycolylmuramic acid, and muramic acid. The carboxylic functions of the peptide side-chains of lipid II showed three types of modification, with the dominant one being amidation. The preferred site for amidation is the free carboxyl group of the meso-diaminopimelic acid residue. Diamidated species were also observed. The carboxylic function of the terminal D alanine of some molecules is methylated, as are all three carboxylic acid functions of other molecules. This study represents the first structural analysis of mycobacterial lipid I and II and the first report of extensive modifications of these molecules. The observation that lipid I was unmodified strongly suggests that the lipid II intermediates of M. smegmatis are substrates for a variety of enzymes that introduce modifications to the sugar and amino acid residues prior to the synthesis of peptidoglycan. PMID- 15805522 TI - Participation of 3'-to-5' exoribonucleases in the turnover of Bacillus subtilis mRNA. AB - Four 3'-to-5' exoribonucleases have been identified in Bacillus subtilis: polynucleotide phosphorylase (PNPase), RNase R, RNase PH, and YhaM. Mutant strains were constructed that were lacking PNPase and one or more of the other three ribonucleases or that had PNPase alone. Analysis of the decay of mRNA encoded by seven small, monocistronic genes showed that PNPase was the major enzyme involved in mRNA turnover. Significant levels of decay intermediates, whose 5' ends were at the transcriptional start site and whose 3' ends were at various positions in the coding sequence, were detected only when PNPase was absent. A detailed analysis of rpsO mRNA decay showed that decay intermediates accumulated as the result of a block to 3'-to-5' processivity at the base of stem loop structures. When RNase R alone was present, it was also capable of degrading mRNA, showing the involvement of this exonuclease in mRNA turnover. The degradative activity of RNase R was impaired when RNase PH or YhaM was also present. Extrapolation from the seven genes examined suggested that a large number of mRNA fragments was present in the PNPase-deficient mutant. Maintenance of the free ribosome pool in this strain would require a high level of activity on the part of the tmRNA trans translation system. A threefold increase in the level of peptide tagging was observed in the PNPase-deficient strain, and selective pressure for increased tmRNA activity was indicated by the emergence of mutant strains with elevated tmRNA transcription. PMID- 15805523 TI - Surprising dependence on postsegregational killing of host cells for maintenance of the large virulence plasmid of Shigella flexneri. AB - Low-copy-number plasmids all encode multiple systems to ensure their propagation, including replication, partition (active segregation), and postsegregational killing (PSK) systems. PSK systems kill those rare cells that lose the plasmid due to replication or segregation errors. PSK systems should not be used as the principle means of maintaining the plasmid. The metabolic cost of killing the many cured cells that would arise from random plasmid segregation is far too high. Here we describe an interesting exception to this rule. Maintenance of the large virulence plasmid of Shigella flexneri is highly dependent on one of its PSK systems, mvp, at 37 degrees C, the temperature experienced during pathogenesis. At 37 degrees C, the plasmid is very unstable and mvp efficiently kills the resulting cured bacterial cells. This imposes a major growth disadvantage on the virulent bacterial population. The systems that normally ensure accurate plasmid replication and segregation are attenuated or overridden at 37 degrees C. At 30 degrees C, a temperature encountered by Shigella in the outside environment, the maintenance systems function normally and the plasmid is no longer dependent on mvp. We discuss why the virulent pathogen tolerates this self-destructive method of propagation at the temperature of infection. PMID- 15805524 TI - Regulation of NAD synthesis by the trifunctional NadR protein of Salmonella enterica. AB - The three activities of NadR were demonstrated in purified protein and assigned to separate domains by missense mutations. The N-terminal domain represses transcription of genes for NAD synthesis and salvage. The C-terminal domain has nicotinamide ribose kinase (NmR-K; EC 2.7.1.22) activity, which is essential for assimilation of NmR, converting it internally to nicotinamide mononucleotide (NMN). The central domain has a weak adenylyltransferase (NMN-AT; EC 2.7.7.1) activity that converts NMN directly to NAD but is physiologically irrelevant. This central domain mediates regulatory effects of NAD on all NadR activities. In the absence of effectors, pure NadR protein binds operator DNA (the default state) and is released by ATP (expected to be present in vivo). NAD allows NadR to bind DNA in the presence of ATP and causes repression in vivo. A superrepressor mutation alters an ATP-binding residue in the central (NMN-AT) domain. This eliminates NMN-AT activity and places the enzyme in its default (DNA binding) state. The mutant protein shows full NmR kinase activity that is 10-fold more sensitive to NAD inhibition than the wild type. It is proposed that NAD and the superrepressor mutation exert their effects by preventing ATP from binding to the central domain. PMID- 15805525 TI - Cross-resistance of Escherichia coli RNA polymerases conferring rifampin resistance to different antibiotics. AB - In this study we further defined the rifampin-binding sites in Escherichia coli RNA polymerase (RNAP) and determined the relationship between rifampin-binding sites and the binding sites of other antibiotics, including two rifamycin derivatives, rifabutin and rifapentine, and streptolydigin and sorangicin A, which are unrelated to rifampin, using a purified in vitro system. We found that there is almost a complete correlation between resistance to rifampin (Rif(r)) and reduced rifampin binding to 12 RNAPs purified from different rpoB Rif(r) mutants and a complete cross-resistance among the different rifamycin derivatives. Most Rif(r) RNAPs were sensitive to streptolydigin, although some exhibited weak resistance to this antibiotic. However, 5 out of the 12 Rif(r) RNAPs were partially resistant to sorangicin A, and one was completely cross resistant to sorangicin A, indicating that the binding site(s) for these two antibiotics overlaps. Both rifampin and sorangicin A inhibited the transition step between transcription initiation and elongation; however, longer abortive initiation products were produced in the presence of the latter, indicating that the binding site for sorangicin A is within the rifampin-binding site. Competition experiments of different antibiotics with (3)H-labeled rifampin for binding to wild-type RNAP further confirmed that the binding sites for rifampin, rifabutin, rifapentine, and sorangicin A are shared, whereas the binding sites for rifampin and streptolydigin are distinct. Because Rif(r) mutations are highly conserved in eubacteria, our results indicate that this set of Rif(r) mutant RNAPs can be used to screen for new antibiotics that will inhibit the growth of Rif(r) pathogenic bacteria. PMID- 15805526 TI - Catabolite repression of the propionate catabolic genes in Escherichia coli and Salmonella enterica: evidence for involvement of the cyclic AMP receptor protein. AB - Previous studies with Salmonella enterica serovar Typhimurium LT2 demonstrated that transcriptional activation of the prpBCDE operon requires the function of transcription factor PrpR, sigma-54, and IHF. In this study, we found that transcription from the prpBCDE and prpR promoters was down-regulated by the addition of glucose or glycerol, indicating that these genes may be regulated by the cyclic AMP (cAMP)-cAMP receptor protein (CRP) complex. Targeted mutagenesis of a putative CRP-binding site in the promoter region between prpR and prpBCDE suggested that these genes are under the control of CRP. Furthermore, cells with defects in cya or crp exhibited reduced transcriptional activation of prpR and prpBCDE in Escherichia coli. These results demonstrate that propionate metabolism is subject to catabolite repression by the global transcriptional regulator CRP and that this regulation is effected through control of both the regulator gene prpR and the prpBCDE operon itself. The unique properties of the regulation of these two divergent promoters may have important implications for mechanisms of CRP-dependent catabolite repression acting in conjunction with a member of the sigma-54 family of transcriptional activators. PMID- 15805527 TI - Antimutator role of DNA glycosylase MutY in pathogenic Neisseria species. AB - Genome alterations due to horizontal gene transfer and stress constantly generate strain on the gene pool of Neisseria meningitidis, the causative agent of meningococcal (MC) disease. The DNA glycosylase MutY of the base excision repair pathway is involved in the protection against oxidative stress. MC MutY expressed in Escherichia coli exhibited base excision activity towards DNA substrates containing A:7,8-dihydro-8-oxo-2'-deoxyguanosine and A:C mismatches. Expression in E. coli fully suppressed the elevated spontaneous mutation rate found in the E. coli mutY mutant. An assessment of MutY activity in lysates of neisserial wild type and mutY mutant strains showed that both MC and gonococcal (GC) MutY is expressed and active in vivo. Strikingly, MC and GC mutY mutants exhibited 60- to 140-fold and 20-fold increases in mutation rates, respectively, compared to the wild-type strains. Moreover, the differences in transitions and transversions in rpoB conferring rifampin resistance observed with the wild type and mutants demonstrated that the neisserial MutY enzyme works in preventing GC-->AT transversions. These findings are important in the context of models linking mutator phenotypes of disease isolates to microbial fitness. PMID- 15805528 TI - Comprehensive characterization of the contribution of individual SigB-dependent general stress genes to stress resistance of Bacillus subtilis. AB - The sigma(B)-dependent general stress regulon of Bacillus subtilis comprises more than 150 members. Induction of this regulon by imposition of environmental or metabolic stress confers multiple, nonspecific, and preemptive stress resistance to nongrowing, nonsporulated cells of B. subtilis. In this study we performed a regulon-wide phenotypic screening analysis to determine the stress sensitivity profiles of 94 mutants defective in candidate members of the general stress regulon that were previously identified in our transcriptional profiling study of the general stress response of B. subtilis. The phenotypic screening analysis included analysis of adaptation to a growth-inhibiting concentration of ethanol (10%, vol/vol) or NaCl (10%, wt/vol), severe heat shock (54 degrees C), and low temperature (survival at 4 degrees C and growth at 12.5 degrees C). Surprisingly, 85% of the mutants tested displayed increased sensitivity at an alpha confidence level of < or =0.01 to at least one of the four stresses tested, and 62% still exhibited increased sensitivity at an alpha of < or =0.001. In essence, we were able to assign 63 genes (28 genes with an alpha of < or =0.001) to survival after ethanol shock, 37 genes (28 genes with an alpha of < or =0.001) to protection from NaCl shock, 34 genes (24 genes with an alpha of < or =0.001) to survival at 4 degrees C, and 10 genes (3 genes with an alpha of < or =0.001) to management of severe heat shock. Interestingly, there was a substantial overlap between the genes necessary for survival during ethanol shock and the genes necessary for survival at 4 degrees C, and there was also an overlap between genes required for survival during ethanol shock and genes required for survival during NaCl shock. Our data provide evidence for the importance of the sigma(B) regulon at low temperatures, not only for growth but also for survival. Moreover, the data imply that a secondary oxidative stress seems to be a common component of the severe stresses tested. PMID- 15805529 TI - Cytochrome oxidase deficiency protects Escherichia coli from cell death but not from filamentation due to thymine deficiency or DNA polymerase inactivation. AB - Temperature-sensitive DNA polymerase mutants (dnaE) are protected from cell death on incubation at nonpermissive temperature by mutation in the cydA gene controlling cytochrome bd oxidase. Protection is observed in complex (Luria Bertani [LB]) medium but not on minimal medium. The cydA mutation protects a thymine-deficient strain from death in the absence of thymine on LB but not on minimal medium. Both dnaE and Deltathy mutants filament under nonpermissive conditions. Filamentation per se is not the cause of cell death, because the dnaE cydA double mutant forms long filaments after 24 h of incubation in LB medium at nonpermissive temperature. These filaments have multiply dispersed nucleoids and produce colonies on return to permissive conditions. The protective effect of a deficiency of cydA at high temperature is itself suppressed by overexpression of cytochrome bo3, indicating that the phenomenon is related to energy metabolism rather than to a specific effect of the cydA protein. We propose that filamentation and cell death resulting from thymine deprivation or slowing of DNA synthesis are not sequential events but occur in response to the same or a similar signal which is modulated in complex medium by cytochrome bd oxidase. The events which follow inhibition of replication fork progression due to either polymerase inactivation, thymine deprivation, or hydroxyurea inhibition differ in detail from those following actual DNA damage. PMID- 15805530 TI - Enhanced synthesis of internalin A in aro mutants of Listeria monocytogenes indicates posttranscriptional control of the inlAB mRNA. AB - Listeria monocytogenes mutants with deletions in aroA, aroB, or aroE exhibited strong posttranscriptional upregulation of internalin A (InlA) and InlB synthesis, which resulted in a more-than-10-fold increase in InlA-mediated internalization by epithelial Caco-2 cells and a 4-fold increase in InlB-mediated internalization by microvascular endothelial cells (human brain microvascular endothelial cells) compared to the wild-type strain. The increase in InlA and InlB production was not due to enhanced PrfA- and/or sigma factor B (SigB) dependent inlAB transcription but was caused by enhanced translation of the inlAB transcripts in the aro mutants. All inlA(B) transcripts had a 396-nucleotide upstream 5' untranslated region (UTR). Different deletions introduced into this UTR led to significant reductions in InlA and InlB synthesis; enhanced translation of all of the truncated transcripts in the aro mutants was, however, still observed. Thus, translation of the inlAB transcripts was subject to two modes of posttranscriptional control, one mediated by the UTR structure and the other mediated by the aro mutation. The latter mode of control seemed to be related to the predominantly anaerobic metabolism of the aro mutants. PMID- 15805531 TI - MinC mutants deficient in MinD- and DicB-mediated cell division inhibition due to loss of interaction with MinD, DicB, or a septal component. AB - The min locus encodes a negative regulatory system that limits formation of the cytokinetic Z ring to midcell by preventing its formation near the poles. Of the three Min proteins, MinC is the inhibitor and prevents Z-ring formation by interacting directly with FtsZ. MinD activates MinC by recruiting it to the membrane and conferring a higher affinity on the MinCD complex for a septal component. MinE regulates the cellular location of MinCD by inducing MinD, and thereby MinC, to oscillate between the poles of the cell, resulting in a time averaged concentration of MinCD on the membrane that is lowest at midcell. MinC can also be activated by the prophage-encoded protein DicB, which targets MinC to the septum without recruiting it first to the membrane. Previous studies have shown that the C-terminal domain of MinC is responsible for the interaction with MinD, DicB, and the septal component. In the present study, we isolated mutations in the C-terminal domain of MinC that affected its interaction with MinD, DicB, and the septal component. Among the mutations isolated, R133A and S134A are specifically deficient in the interaction with MinD, E156A is primarily affected in the interaction with DicB, and R172A is primarily deficient in the interaction with the septum. These mutations differentiate the interactions of MinC with its partners and further support the model of MinCD- and MinC-DicB-mediated cell division inhibition. PMID- 15805532 TI - Genetic and structural analysis of the Bacteroides conjugative transposon CTn341. AB - The genetic structure and functional organization of a Bacteroides conjugative transposon (CTn), CTn341, were determined. CTn341 was originally isolated from a tetracycline-resistant clinical isolate of Bacteroides vulgatus. The element was 51,993 bp long, which included a 5-bp coupling sequence that linked the transposon ends in the circular form. There were 46 genes, and the corresponding gene products fell into three major functional groups: DNA metabolism, regulation and antibiotic resistance, and conjugation. The G + C content and codon usage observed in the functional groups suggested that the groups belong to different genetic lineages, indicating that CTn341 is a composite, modular element. Mutational analysis of genes representing the different functional groups provided evidence for the gene assignments and showed that the basic conjugation and excision genes are conserved among Bacteroides spp. A group IIA1 intron, designated B.f.I1, was found to be inserted into the bmhA methylase gene. Reverse transcriptase PCR analysis of CTn341 RNA showed that B.fr.I1 was functional and was spliced out of the bmhA gene. Six related CTn-like elements were found in the genome sequences of Bacteroides fragilis NCTC9343 and Bacteroides thetaiotaomicron VPI5482. The putative elements were similar to CTn341 primarily in the tra and mob regions and in the exc gene, and several appeared to contain intron elements. Our data provide the first reported sequence for a complete Bacteroides CTn, and they should be of considerable benefit to further functional and genetic analyses of antibiotic resistance elements and genome evolution in Bacteroides. PMID- 15805533 TI - Characterization of type 2 quorum sensing in Klebsiella pneumoniae and relationship with biofilm formation. AB - Quorum sensing is a process by which bacteria communicate by using secreted chemical signaling molecules called autoinducers. Many bacterial species modulate the expression of a wide variety of physiological functions in response to changes in population density by this mechanism. In this study, the opportunistic pathogen Klebsiella pneumoniae was observed to secrete type 2 signaling molecules. A homologue of luxS, the gene required for AI-2 synthesis in Vibrio harveyi, was isolated from the K. pneumoniae genome. A V. harveyi bioassay showed the luxS functionality in K. pneumoniae and its ability to complement the luxS negative phenotype of Escherichia coli DH5alpha. Autoinducer activity was detected in the supernatant, and maximum expression of specific messengers detected by quantitative reverse transcription-PCR analysis occurred during the late exponential phase. The highest levels of AI-2 were observed in minimal medium supplemented with glycerol. To determine the potential role of luxS in colonization processes, a K. pneumoniae luxS isogenic mutant was constructed and tested for its capacity to form biofilms in vitro on an abiotic surface and to colonize the intestinal tract in a murine model. No difference was observed in the level of intestinal colonization between the wild-type strain and the luxS mutant. Microscopic analysis of biofilm structures revealed that the luxS mutant was able to form a mature biofilm but with reduced capacities in the development of microcolonies, mostly in the early steps of biofilm formation. These data suggest that a LuxS-dependent signal plays a role in the early stages of biofilm formation by K. pneumoniae. PMID- 15805534 TI - Polarity of enteropathogenic Escherichia coli EspA filament assembly and protein secretion. AB - Type III secretion systems (TTSS) are sophisticated macromolecular structures that play an imperative role in bacterial infections and human disease. The TTSS needle complex is conserved among bacterial pathogens and shows broad similarity to the flagellar basal body. However, the TTSS of enteropathogenic and enterohemorrhagic Escherichia coli, two important human enteric pathogens, is unique in that it has an approximately 12-nm-diameter filamentous extension to the needle that is composed of the secreted translocator protein EspA. EspA filaments and flagellar structures have very similar helical symmetry parameters. In this study we investigated EspA filament assembly and the delivery of effector proteins across the bacterial cell wall. We show that EspA filaments are elongated by addition of EspA subunits to the tip of the growing filament. Moreover, EspA filament length is modulated by the availability of intracellular EspA subunits. Finally, we provide direct evidence that EspA filaments are hollow conduits through which effector proteins are delivered to the extremity of the bacterial cell (and subsequently into the host cell). PMID- 15805535 TI - Crystal structure of the flagellar rotor protein FliN from Thermotoga maritima. AB - FliN is a component of the bacterial flagellum that is present at levels of more than 100 copies and forms the bulk of the C ring, a drum-shaped structure at the inner end of the basal body. FliN interacts with FliG and FliM to form the rotor mounted switch complex that controls clockwise-counterclockwise switching of the motor. In addition to its functions in motor rotation and switching, FliN is thought to have a role in the export of proteins that form the exterior structures of the flagellum (the rod, hook, and filament). Here, we describe the crystal structure of most of the FliN protein of Thermotoga maritima. FliN is a tightly intertwined dimer composed mostly of beta sheet. Several well-conserved hydrophobic residues form a nonpolar patch on the surface of the molecule. A mutation in the hydrophobic patch affected both flagellar assembly and switching, showing that this surface feature is important for FliN function. The association state of FliN in solution was studied by analytical ultracentrifugation, which provided clues to the higher-level organization of the protein. T. maritima FliN is primarily a dimer in solution, and T. maritima FliN and FliM together form a stable FliM(1)-FliN(4) complex. Escherichia coli FliN forms a stable tetramer in solution. The arrangement of FliN subunits in the tetramer was modeled by reference to the crystal structure of tetrameric HrcQB(C), a related protein that functions in virulence factor secretion in Pseudomonas syringae. The modeled tetramer is elongated, with approximate dimensions of 110 by 40 by 35 Angstroms, and it has a large hydrophobic cleft formed from the hydrophobic patches on the dimers. On the basis of the present data and available electron microscopic images, we propose a model for the organization of FliN subunits in the C ring. PMID- 15805536 TI - Role of the Rep helicase gene in homologous recombination in Neisseria gonorrhoeae. AB - In Escherichia coli, the Rep helicase has been implicated in replication fork progression, replication restart, homologous recombination, and DNA repair. We show that a Neisseria gonorrhoeae rep mutant is deficient in the homologous recombination-mediated processes of DNA transformation and pilus-based colony variation but not in DNA repair. PMID- 15805537 TI - Functional reassembly of the Escherichia coli maltose transporter following purification of a MalF-MalG subassembly. AB - Taking advantage of a chaperone-like function of MalK, a stable complex of MalF MalG could be solubilized from the Escherichia coli membrane and purified in high yield in the absence of MalK. This MalF-MalG complex was competent for efficient reassembly of a functional MalFGK(2) maltose transporter complex both in detergent solution and in proteoliposomes. PMID- 15805538 TI - Identification of rcnA (yohM), a nickel and cobalt resistance gene in Escherichia coli. AB - We report here on the isolation and primary characterization of the yohM gene of Escherichia coli. We show that yohM encodes a membrane-bound polypeptide conferring increased nickel and cobalt resistance in E. coli. yohM was specifically induced by nickel or cobalt but not by cadmium, zinc, or copper. Mutation of yohM increased the accumulation of nickel inside the cell, whereas cells harboring yohM in multicopy displayed reduced intracellular nickel content. Our data support the hypothesis that YohM is the first described efflux system for nickel and cobalt in E. coli. We propose rcnA (resistance to cobalt and nickel) as the new denomination of yohM. PMID- 15805539 TI - Clonal distribution and phase-variable expression of a major histocompatibility complex analogue protein in Staphylococcus aureus. AB - The mapW gene of Staphylococcus aureus strain N315 contains a poly(A) tract which truncates translation of the protein. This study demonstrates that mapW is an allelic variant of the map/eap genes found in other strains and that the variation in the length of this poly(A) tract suggests that it is a contingency locus. PMID- 15805540 TI - Positive selection for loss-of-function tat mutations identifies critical residues required for TatA activity. AB - The Tat system, found in the cytoplasmic membrane of many bacteria, is a general export pathway for folded proteins. Here we describe the development of a method, based on the transport of chloramphenicol acetyltransferase, that allows positive selection of mutants defective in Tat function. We have demonstrated the utility of this method by selecting novel loss-of-function alleles of tatA from a pool of random tatA mutations. Most of the mutations that were isolated fall in the amphipathic region of TatA, emphasizing the pivotal role that this part of the protein plays in TatA function. PMID- 15805541 TI - Role of ceramide in Ca2+-sensing receptor-induced apoptosis. AB - Increased extracellular Ca(2+) ([Ca(2+)](o)) can damage tissues, but the molecular mechanisms by which this occurs are poorly defined. Using HEK 293 cell lines that stably overexpress the Ca(2+)-sensing receptor (CaR), a G protein coupled receptor, we demonstrate that activation of the CaR leads to apoptosis, which was determined by nuclear condensation, DNA fragmentation, caspase-3 activation, and increased cytosolic cytochrome c. This CaR-induced apoptotic pathway is initiated by CaR-induced accumulation of ceramide which plays an important role in inducing cell death signals by distinct G protein-independent signaling pathways. Pretreatment of wild-type CaR-expressing cells with pertussis toxin inhibited CaR-induced [(3)H]ceramide formation, c-Jun phosphorylation, and caspase-3 activation. The ceramide accumulation, c-Jun phosphorylation, and caspase-3 activation by the CaR can be abolished by sphingomyelinase and ceramide synthase inhibitors in different time frames. Cells that express a nonfunctional mutant CaR that were exposed to the same levels of [Ca(2+)](o) showed no evidence of activation of the apoptotic pathway. In conclusion, we report the involvement of the CaR in stimulating programmed cell death via a pathway involving GTP binding protein alpha subunit (Galpha(i))-dependent ceramide accumulation, activation of stress-activated protein kinase/c-Jun N-terminal kinase, c-Jun phosphorylation, caspase-3 activation, and DNA cleavage. PMID- 15805543 TI - ACAT2 contributes cholesteryl esters to newly secreted VLDL, whereas LCAT adds cholesteryl ester to LDL in mice. AB - The relative contributions of ACAT2 and LCAT to the cholesteryl ester (CE) content of VLDL and LDL were measured. ACAT2 deficiency led to a significant decrease in the percentage of CE (37.2 +/- 2.1% vs. 3.9 +/- 0.8%) in plasma VLDL, with a concomitant increase in the percentage of triglyceride (33.0 +/- 3.2% vs. 66.7 +/- 2.5%). Interestingly, the absence of ACAT2 had no apparent effect on the percentage CE in LDL, whereas LCAT deficiency significantly decreased the CE percentage (38.6 +/- 4.0% vs. 54.6 +/- 1.9%) and significantly increased the phospholipid percentage (11.2 +/- 0.9% vs. 19.3 +/- 0.1%) of LDL. When both LCAT and ACAT2 were deficient, VLDL composition was similar to VLDL of the ACAT2 deficient mouse, whereas LDL was depleted in core lipids and enriched in surface lipids, appearing discoidal when observed by electron microscopy. We conclude that ACAT2 is important in the synthesis of VLDL CE, whereas LCAT is important in remodeling VLDL to LDL. Liver perfusions were performed, and perfusate apolipoprotein B accumulation rates in ACAT2-deficient mice were not significantly different from those of controls; perfusate VLDL CE decreased from 8.0 +/- 0.8% in controls to 0 +/- 0.7% in ACAT2-deficient mice. In conclusion, our data establish that ACAT2 provides core CE of newly secreted VLDL, whereas LCAT adds CE during LDL particle formation. PMID- 15805542 TI - Monolysocardiolipins accumulate in Barth syndrome but do not lead to enhanced apoptosis. AB - Barth syndrome (BTHS) is an X-linked recessive disorder that is biochemically characterized by low cellular levels of the mitochondrial phospholipid cardiolipin (CL). Previously, we discovered that the yeast disruptant of the TAZ ortholog in Saccharomyces cerevisiae not only displays CL deficiency but also accumulates monolysocardiolipins (MLCLs), which are intermediates in CL remodeling. Therefore, we set out to investigate whether MLCL accumulation also occurs in BTHS. Indeed, we observed MLCL accumulation in heart, muscle, lymphocytes, and cultured lymphoblasts of BTHS patients; however, only very low levels of these lysophospholipids were found in platelets and fibroblasts of these patients. Although the fatty acid composition of the MLCLs was different depending on the tissue source, it did parallel the fatty acid composition of the (remaining) CLs. The possible implications of these findings for the two reported CL remodeling mechanisms, transacylation and deacylation/reacylation, are discussed. Because MLCLs have been proposed to be involved in the initiation of apoptosome-mediated cell death by the sequestration of the proapoptotic protein (t)BH3-interacting domain death agonist (Bid) to the mitochondrial membrane, we used control and BTHS lymphoblasts to investigate whether the accumulation of MLCLs results in higher levels of apoptosis. We found no differences in susceptibility to death receptor-mediated apoptosis or in cellular distribution of Bid, cytochrome c, and other parameters, implying that MLCL accumulation does not lead to enhanced apoptosis in cultured BTHS lymphoblasts. PMID- 15805544 TI - A redox-sensitive pathway mediates oxidized LDL-induced downregulation of insulin like growth factor-1 receptor. AB - Oxidized low density lipoprotein (OxLDL) has multiple proatherogenic effects, including induction of apoptosis. We have recently shown that OxLDL markedly downregulates insulin-like growth factor-1 receptor (IGF-1R) in human aortic smooth muscle cells, and that IGF-1R overexpression blocks OxLDL-induced apoptosis. We hypothesized that specific OxLDL-triggered signaling events led to IGF-1R downregulation and apoptosis. We examined OxLDL signaling pathways and found that neither IGF-1R downregulation nor the proapoptotic effect was blocked by inhibition of OxLDL-triggered extracellular signal-regulated kinase, p38 mitogen-activated protein kinase (MAPK), or peroxisome proliferator-activated receptor gamma (PPARgamma) signaling pathways, as assessed using specific inhibitors. However, antioxidants, polyethylene glycol catalase, superoxide dismutase, and Trolox completely blocked OxLDL downregulation of IGF-1R and OxLDL induced apoptosis. Nordihydroguaiaretic acid, AA-861, and baicalein, which are lipoxygenase inhibitors and also have antioxidant activity, blocked IGF-1R downregulation and apoptosis as well as reactive oxygen species (ROS) production. These results suggest that OxLDL enhances ROS production possibly through lipoxygenase activity, leading to IGF-1R downregulation and apoptosis. Furthermore, anti-CD36 scavenger receptor antibody markedly inhibited OxLDL induced IGF-1R downregulation and apoptosis as well as ROS production. In conclusion, our data demonstrate that OxLDL downregulates IGF-1R via redox sensitive pathways that are distinct from OxLDL signaling through MAPK- and PPARgamma-involved pathways but may involve a CD36-dependent mechanism. PMID- 15805545 TI - Changes in gene expression associated with loss of function of the NSDHL sterol dehydrogenase in mouse embryonic fibroblasts. AB - Seven human disorders of postsqualene cholesterol biosynthesis have been described. One of these, congenital hemidysplasia with ichthyosiform nevus and limb defects (CHILD) syndrome, results from mutations in the X-linked gene NADH sterol dehydrogenase-like (NSDHL) encoding a sterol dehydrogenase. A series of mutant alleles of the murine Nsdhl gene are carried by bare patches (Bpa) mice, with Bpa(1H) representing a null allele. Heterozygous Bpa(1H) females display skin and skeletal abnormalities in a distribution reflecting random X inactivation, whereas hemizygous male embryos die before embryonic day 10.5. To investigate the molecular basis of defects associated with perturbations in cholesterol biosynthesis, microarray analysis was performed comparing gene expression in embryonic fibroblasts expressing the Bpa(1H) allele versus wild type (wt) cells. Labeled cDNAs from cells grown in normal serum or lipid-depleted serum (LDS) were hybridized to microarrays containing 22,000 mouse genes. Among 44 genes that showed higher expression in the Bpa(1H) versus wt cells grown in LDS, 11 function in cholesterol biosynthesis, 7 are involved in fatty acid synthesis, 3 (Srebp2, Insig1, and Orf11) encode sterol-regulatory proteins, and 2 (Ldlr and StarD4) are lipid transporters. Of the 21 remaining genes, 16 are known genes, some of which have been implicated previously in cholesterol homeostasis or lipid-mediated signaling, and 5 are uncharacterized cDNA clones. PMID- 15805547 TI - The recycling of apolipoprotein E in macrophages: influence of HDL and apolipoprotein A-I. AB - The ability of apolipoprotein E (apoE) to be spared degradation in lysosomes and to recycle to the cell surface has been demonstrated by our group and others, but its physiologic relevance is unknown. In this study, we characterized apoE recycling in primary murine macrophages and probed the effects of HDL and apoA-I on this process. In cells pulsed with (125)I.apoE bound to VLDL, intact apoE was found in the chase medium for up to 24 h after the pulse. Approximately 27 +/- 5% of the apoE internalized during the pulse was recycled after 4 h of chase. Addition of apoA-I and HDL increased apoE recycling to 45 +/- 3% and 46 +/- 3%, respectively, similar to the amount of apoE recycled after pulsing the cells with (125)I.apoE.HDL. In addition, apoA-I-producing macrophages from transgenic mice showed increased apoE recycling at 4 h (38 +/- 3%). Increased ABCA1 expression potentiated apoE recycling, suggesting that recycling occurs via ABCA1. Finally, in the presence of apoA-I, recycled apoE exited the cells on HDL-like particles. These results suggest that apoE recycling in macrophages may be part of a larger signaling loop activated by HDL and directed at maximizing cholesterol losses from the cell. PMID- 15805546 TI - Chemical modification of proteins during peroxidation of phospholipids. AB - Chemical modification of proteins by advanced glycation and lipoxidation end products is implicated in the pathogenesis of macrovascular disease in aging and diabetes. To identify biomarkers of the lipoxidative modification of protein, we studied the oxidation of phospholipids in the presence of the model protein RNase A and compared protein-bound products formed in these reactions with those formed during oxidation of plasma proteins. Metal-catalyzed oxidation of 1-palmitoyl-2 arachidonoyl-phosphatidylcholine or 1-palmitoyl-2-linoleoyl-phosphatidylcholine in the presence of RNase led to the loss of amino groups in RNase and the incorporation of phosphate, hexanoate, pentanedioate, nonanedioate, and palmitate into protein. Protein-bound palmitate and phosphate correlated strongly with one another, and protein-bound pentanedioate and nonanedioate, derived from arachidonate and linoleate, respectively, accounted for approximately 20% of the cross-linking of lipid phosphorus to protein. Similar results were obtained on oxidation of total plasma or isolated LDL. We conclude that alkanedioic acids are quantitatively important linkers of oxidized phospholipids to proteins and that measurement of protein-bound phosphate and long-chain fatty acids may be useful for assessing long-term lipid peroxidative damage to proteins in vivo. Analyses of plasma proteins from control and diabetic patients indicated significant increases in lipoxidative modification of protein in diabetic compared with control subjects. PMID- 15805548 TI - Cysteine mutants of human apolipoprotein A-I: a study of secondary structural and functional properties. AB - Apolipoprotein A-I(Milano) (A-I(M)) (R173C), a natural mutant of human apolipoprotein A-I (apoA-I), and five other cysteine variants of apoA-I at residues 52 (S52C), 74 (N74C), 107 (K107C), 129 (G129C), and 195 (K195C) were generated. Cysteine residues were incorporated in each of the various helices at the same helical wheel position as for the substitution in A-I(M). The secondary structural properties of the monomeric mutants, their abilities to bind lipid and to promote cholesterol efflux from THP-1 macrophages, and the possibility of antiperoxidation were investigated. Results showed that the alpha helical contents of all of the cysteine mutants were similar to that of wild-type apoA-I (wtapoA-I). The cysteine variant of A-I(M) at residue 173 [A-I(M)(R173C)] exhibited weakened structural stability, whereas A-I(G129C) a more stable structure than wtapoA-I. A-I(G129C) and A-I(K195C) exhibited significantly impaired capabilities to bind lipid compared with wtapoA-I. A-I(K107C) possessed a higher capacity to promote cholesterol efflux from macrophages than wtapoA-I, and A-I(M)(R173C) and A-I(K195C) exhibited an impaired efflux capability. Neither A-I(M)(R173C) nor any other cysteine mutant could resist oxidation against lipoxygenase. In summary, in spite of the similar mutant position on the helix, these variants exhibited different structural features or biological activities, suggesting the potential influence of the local environment of mutations on the whole polypeptide chain. PMID- 15805549 TI - Genome-wide linkage analyses and candidate gene fine mapping for HDL3 cholesterol: the Framingham Study. AB - High density lipoprotein cholesterol (HDL-C) is inversely associated with coronary heart disease and has a genetic component; however, linkage to HDL-C is not conclusive. Subfractions of HDL, such as HDL(3)-C, may be better phenotypes for linkage studies. Using HDL(3)-C levels measured on 907 Framingham Heart Study subjects from 330 families around 1987, we conducted a genome-wide variance components linkage analysis with 401 microsatellite markers spaced approximately 10 centimorgan (cM) apart. Nine candidate genes were identified and annotated using a bioinformatics approach in the region of the highest linkage peak. Twenty eight single nucleotide polymorphisms (SNPs) were selected from these candidate genes, and linkage and family-based association fine mapping were conducted using these SNPs. The highest multipoint log-of-the-odds (LOD) score from the initial linkage analysis was 3.7 at 133 cM on chromosome 6. Linkage analyses with additional SNPs yielded the highest LOD score of 4.0 at 129 cM on chromosome 6. Family-based association analysis revealed that SNP rs2257104 in PLAGL1 at approximately 143 cM was associated with multivariable adjusted HDL(3) (P = 0.03). Further study of the linkage region and exploration of other variants in PLAGL1 are warranted to define the potential functional variants of HDL-C metabolism. PMID- 15805550 TI - Fluorescent leukotriene B4: potential applications. AB - Leukotriene B4 (LTB4) is a potent lipid mediator of inflammation that acts primarily via a seven-transmembrane-spanning, G-protein-coupled receptor denoted BLT1. Here, we describe the synthesis and characterization of fluorescent analogs of LTB4 that are easy to produce, inexpensive, and without the disadvantages of a radioligand. Fluorescent LTB4 is useful for labeling LTB4 receptors for which no antibodies are available and for performing one-step fluorescence polarization assays conducive to high-throughput screening. We found that orange and green fluorescent LTB4 were full agonists that activated the LTB4 receptor BLT1 with EC50 values of 68 and 40 nM, respectively (4.5 nM for unmodified LTB4). Flow cytometric measurements and confocal imaging showed that fluorescent LTB4 colocalized with BLT1. Fluorescence polarization measurements showed that orange fluorescent LTB4 bound to BLT1 with a Kd of 66 nM and that this binding could be displaced by unlabeled LTB4 and other BLT1-specific ligands. Fluorescent LTB4 analogs were also able to displace tritiated LTB4. Orange fluorescent LTB4 binding to enhanced green fluorescent protein-tagged BLT1 could be observed using fluorescence resonance energy transfer. In addition to being a useful alternative to radiolabeled LTB4, the unique properties of fluorescently labeled LTB4 allow a variety of detection technologies to be used. PMID- 15805551 TI - Increased arachidonic acid concentration in the brain of Flinders Sensitive Line rats, an animal model of depression. AB - Depression may be associated with impaired membrane PUFA composition, especially decreased n-3 PUFA. This assumption has not been tested at the level of brain tissue. Moreover, most studies were confounded by dietary variability. We examined the FA composition of selected brain areas in an animal model of depression, the Flinders Sensitive Line (FSL) rat, and compared the findings with those in controls fed identical diets. In all brain regions studied, the concentration of arachidonic acid (AA) was significantly higher in the FSL rats: in the hypothalamus by 21%, in the nucleus accumbens by 24%, in the prefrontal cortex by 31%, and in the striatum by 23%. No significant differences were observed for n-3 PUFA or for the saturated and monounsaturated FAs. Our results confirm the existence of altered brain PUFA composition in an animal model of depression. The finding of increased AA, an n-6 PUFA, rather than decreased n-3 PUFA, emphasizes the importance of both PUFA families in the pathophysiological processes underlying depression. The FSL rat is a useful tool for further elucidation of the FA disturbances in depression. PMID- 15805552 TI - Taking the sting out of needles. PMID- 15805553 TI - Management of chemically contaminated bodies. PMID- 15805555 TI - 'Referral into a void': opinions of general practitioners and others on single point of access to mental health care. AB - In the past, British general practitioners (GPs) have referred patients to individual consultants. There is now a trend towards generic referral, whereby the specialist team is consulted via a single point of access. We examined the impact of this innovation on the relationship between GPs and specialists in mental health care. Sixteen groups of randomly selected GPs and mental health professionals (MHPs) in England discussed clinical scenarios involving the use of mental health interventions for patients with functional somatic symptoms. The meetings were audiotaped, transcribed and analysed to the point at which no major new themes were emerging. The final analysis was confined to a purposive sample of six of the sixteen groups, comprising 54 GPs and 15 MHPs. Although the rationale for the single point of access was clear to both GPs and MHPs, the approach erected boundaries because it impeded the establishment of professional relationships and transfer of knowledge. GPs thought the system reduced their capacity to be accountable to their patients and limited the potential for their own professional development. MHPs did not seem to be aware of GPs' concerns. Effective interprofessional management of individual patients depends upon confidence in colleagues' skills and good communication. Factors that hamper these must be addressed in the development of this system of referral. PMID- 15805554 TI - Therapeutic monoclonal antibodies in oncology. PMID- 15805556 TI - Local anaesthesia for venous cannulation and arterial blood gas sampling: are doctors using it? AB - The pain of venous cannulation and arterial puncture can be greatly lessened by local anaesthesia. We sought information about the use of local anaesthesia for these procedures by doctors working in medicine, surgery and anaesthetics. A questionnaire was hand-delivered to 178 doctors in eight hospitals, all of whom responded. For insertion of large-bore cannulae, local anaesthesia was used by all the anaesthetists but less than half the medical and surgical doctors. For arterial blood sampling it was used by 60% of anaesthetists and 2% of ward doctors. Previous recommendations to use local anaesthesia seem to have been ignored, and in many instances these procedures are more painful than necessary. PMID- 15805557 TI - Portal vein thrombosis despite anticoagulation in a person with diabetes. PMID- 15805558 TI - Perindopril and pulmonary eosinophilic syndrome. PMID- 15805559 TI - Renal bone disease. PMID- 15805560 TI - Fibrinous peritonitis in a patient with rheumatoid arthritis. PMID- 15805561 TI - Two cases of hypothalamic-pituitary sarcoidosis. PMID- 15805562 TI - Giant colonic diverticulum. PMID- 15805563 TI - Human rights and medical ethics. PMID- 15805564 TI - Surviving a clinical exam: a guide for candidates. PMID- 15805565 TI - Parkinson's disease before Parkinson, Vilnius 1814. PMID- 15805566 TI - In Sri Lanka after the tsunami. PMID- 15805567 TI - 35 years with an uninvited guest. PMID- 15805568 TI - MRSA. PMID- 15805569 TI - Good teachers: substainable models. PMID- 15805570 TI - Empowerment for patients with medically unexplained symptoms. PMID- 15805571 TI - BKV-DNA and JCV-DNA co-quantification assay to evaluate viral load in urine and serum. AB - Infections from human polyomaviruses BK and JC (BKV and JCV) occur independently, but concomitant infections and the simultaneous persistence of both viruses have been observed in renal transplant recipients. Several studies have disclosed a correlation between BKV and interstitial nephritis in renal transplant recipients, and an association between JCV and some cases of nephropathy has recently been hypothesized. This article describes the development of a semiquantitative-nested polymerase chain reaction (PCR) assay to simultaneously detect BKV and JCV viral load in urine and serum. The first-round amplification step uses primers that amplify a 385-bp DNA fragment from the "large T antigen" region of both viruses. Samples testing positive in the first step are then run in the second step. In the second-round amplification, different inner primers are used to separately quantify BKV-DNA and/or JCV-DNA. The assay offers several advantages including: (1) rapid submission of clinical samples to screening; (2) verification of the absence of Taq polymerase inhibitors with the use of an internal control; (3) a sensitivity threshold of 10 copies/reaction; and (4) assay running is less labor intensive, cheap, and easy to perform. The assay may be easily used to monitor viral loads versus baseline levels in urine and serum samples from renal transplant recipients to detect those at risk of BKV- or JCV related nephropathy, and to monitor their response to immunosuppression reduction therapy if it occurs. PMID- 15805572 TI - Optimization of high-efficiency transfection of adult human mesenchymal stem cells in vitro. AB - With the advent of recent protocols to isolate multipotent human mesenchymal stem cells (MSCs), there is a need for efficient transfection methodologies for these cells. Most standard transfection methods yield poor transfection efficiencies for MSCs (<1%). Here we have optimized a high-efficiency transfection technique for low passage MSCs derived from adult human bone marrow. This technique is an extension of electroporation, termed amaxa Nucleofection, where plasmid DNA is transfected directly into the cell nucleus, independent of the growth state of the cell. With this technique, we demonstrate up to 90% transfection efficiency of the viable population of MSCs, using plasmid construct containing a standard cytomegalovirus (CMV) early promoter driving expression of green fluorescent protein (GFP). Although little variation in transfection efficiency was observed between patient samples, a 2-fold difference in transfection efficiency and a 10 fold difference in expression levels per cell were seen using two distinct CMV GFP expression plasmids. By fluorescence-activated cell sorting, the GFP expressing cells were sorted and subcultured. At 2 wk posttransfection, approx 25% of the population of sorted cells were GFP positive, and by 3 wk, nearly 10% of the cells still retained GFP expression. Transfection of these cells with plasmid containing either the collagen type I (Col1a1) promoter or the cartilage oligomeric matrix protein (COMP) promoter, each driving expression of GFP, produced a somewhat lower transfection efficiency (approx 40%), due in part to the lower activity of transcription from these promoters compared to that of CMV. Transfection with the collagen type II (Col2a1) promoter linked to GFP exhibited low expression, due to the fact that collagen type II is not expressed in these cells. Upon culturing of the Col2a1-GFP transfected cells in a transforming growth factor-beta3-containing medium known to induce mesenchymal chondrogensis, a significant enhancement of GFP level was seen, indicating the ability of the transfected cells to differentiate into chondrocytes and express cartilage specific genes, such as Col2a1. Taken together, these data provide evidence of the applicability of this technique for the efficient transfection of MSCs. PMID- 15805573 TI - INSULT: a novel mutagenesis method generating high yields of closed circular mutant DNA with one primer per mutant. AB - INSULT, a novel method for the creation of insertions, deletions, and point mutations without subcloning, requires only one new primer per mutant, and produces circular plasmids, obviating the need for special "ultracompetent" cells. The method includes cycles of linear amplification with a thermophilic polymerase, and nick repair after each cycle with a thermophilic ligase. After production of multiple single-stranded copies of circular mutation-bearing plasmid DNA, addition of a "generic" primer followed by one or more polymerase reaction cycles generates double-stranded circular DNA bearing the desired mutation. PMID- 15805574 TI - Identification of salt stress-induced transcripts in potato leaves by cDNA-AFLP. AB - Potato (Solanum tuberosum L.) is highly sensitive to salt stress, which is one of the most important factors limiting plant cultivation. The investigation of plant response to high salinity was envisaged in this report using cDNA-amplified fragment length polymorphism (AFLP). This technique was applied to salt- stressed and control potato plants (cv. Nicola). The expression profiles showed approx 5000 bands. Of these, 154 were upregulated and 120 were repressed by salt stress. In this study we have only considered cDNA fragments that seem to be originated from salt-induced mRNA. Eighteen fragments were then reamplified, cloned, and sequenced. Sequence comparison of these cDNA, identified in response to salt stress in potato, revealed that some of them present homologies with proteins in other species that are involved in cell wall structure and turnover such as proline-rich proteins and beta-galactosidase. A number of identified clones encoded putative stress response proteins such as NADP-dependant glyceraldehyde dehydrogenase and wound-induced protein. In addition, some of them encode proteins related to hypersensitive response against pathogens such as putative late blight and nematode as well as putative pathogenesis-related proteins. These cDNA seem to be differentially expressed in the presence of salt stress as shown by Northern blot or reverse Northern hybridization experiments. PMID- 15805575 TI - Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity. AB - We adapted a previously described Agrobacterium-mediated transient expression system to test the expression level of three constructs carrying the surface antigen 1 (SAG1) of Toxoplasma gondii. Two constructs were based in a Potato virus X (PVX) amplicon. In one of them, the PVX movement protein genes were replaced by the sag1 gene. In the other, the sag1 gene was placed under the control of an additional coat protein subgenomic promoter. In the third construct, the sag1 gene was fused to an apoplastic peptide signal under the CaMV 35S promoter. Western blot analysis of leaf extracts infiltrated with each construct revealed a protein of 35 kDa. SAG1 accumulation in leaves ranged from 0.1 to 0.06% of total soluble protein (equivalent to 10 microg and 6 microg of SAG1 per gram of fresh leaf tissue, respectively). Three of five human seropositive samples reacted with tobacco-expressed SAG1 in Western blot analysis. The C3H mice were immunized with SAG-expressing leaf extracts and perorally challenged with a nonlethal dose of the T. gondii Me49 strain. Mice vaccinated with SAG1 showed significantly lower brain cyst burdens compared to those from the control group. Immunization with SAG1-expressing leaves elicited a specific humoral response with predominant participation of type IgG2a. In conclusion, a functional SAG1 version could be transiently expressed in tobacco leaves. PMID- 15805576 TI - Method for recovery of intact DNA for community analysis of marine intertidal microbial biofilms. AB - A protocol is described for rapid DNA isolation from marine biofilm microorganisms embedded in large amounts of exopolysaccharides. The method is a modification of the hot phenol protocol used for plants tissues, where nonexpensive and easily available enzymes were used. The method is based on the incubation of biofilm biomass samples in an extraction buffer mixed with phenol preheated at 65 degrees C. The procedure can be completed in 2 h and up to 20 samples can be processed simultaneously with ease and DNA of excellent quality, as shown by successfully amplification of polymerase chain reaction (PCR) products. DNA was recovered from a range of intertidal marine biofilms with varying amounts of exopolysaccharides. PMID- 15805577 TI - Molecular biology of the blood-brain barrier. AB - Molecular biological investigations into the brain capillary endothelium and microvasculature, which forms the blood-brain barrier (BBB) in vivo, can provide the platform for the discovery and the molecular cloning of BBB-specific genes. Novel BBB genes can be discovered with either a genomics-based approach such as subtractive suppressive hybridization, or a proteomics approach using subtractive antibody expression cloning. BBB-specific genes are disproportionately transporter genes encoding either for carrier-mediated transporters, active efflux transporters, or receptor-mediated transporters. The discovery of new BBB transporters can lead to the development of new approaches to brain drug delivery using endogenous brain endothelial transporters. PMID- 15805578 TI - Introduction to the background, principles, and state of the art in suicide gene therapy. AB - Gene therapy is defined as a technology that aims to modify the genetic component of cells to gain therapeutic benefits. Suicide gene therapy (or gene-directed enzyme prodrug therapy [GDEPT]) is a two-step treatment for cancer (especially, solid tumors). In the first step, a gene for a foreign enzyme is delivered to the tumor by a vector. Following the expression of the foreign enzyme, a prodrug is administered during the second step, which is selectively activated in the tumor. This article discusses the principles and the theorectical background of GDEPT. A special emphasis is put on enzyme/prodrug systems developed for GDEPT, the design of prodrugs and the kinetic of their activation, the types and the mechanisms of bystander effect and its immunological implications. The possible strategies to improve GDEPT are also discussed. PMID- 15805579 TI - Androgen-receptor coregulators mediate the suppressive effect of androgen signals on vitamin D receptor activity. AB - Overexpression of androgen receptors (AR) in PC-3 cell, and treatment of 5alpha dihydrotestosterone in LNCaP cells lead to the suppression of VDR transactivation. Competition for shared coregulators between AR and VDR is one possible mechanism to explain the suppressive effect of androgen-AR signals on VDR activity. Among the AR coregulators we tested, ARA54, ARA70, supervillin, and gelsolin were found to enhance VDR transactivation. Further characterization of the interaction between ARA54 or ARA70 and VDR demonstrated a direct interaction between VDR and ARA70, but no association between ARA54 and VDR. The LXXLL motif of ARA70 is essential for interaction with VDR and partially responsible for its function as a coactivator of VDR. The suppression of VDR transactivation by AR signal was restored by overexpression of ARA70, but not ARA54. Together, ARA70 and ARA54 modulate VDR transactivation, and the competition for ARA70 mediates the suppressive effect of androgen-AR on VDR transactivation. PMID- 15805580 TI - Effect of growth hormone and estrogen administration on hepatocyte alterations in old ovariectomized female wistar rats. AB - Aging could be due to the accumulation of oxidative damage. On the other hand, growth hormone (GH) and estrogen deficiency induce deleterious effects on different tissues, and hormonal replacement could counteract these effects. We have investigated whether GH and estrogen administration modify some parameters related to oxidative stress and inflammation in hepatocytes isolated from old ovariectomized female rats. Twenty-two month-old ovariectomized animals were divided into control rats, rats treated with GH, rats treated with estradiol, and rats treated with GH+estradiol. Two-month-old intact female rats were used as young reference group. Hepatocytes were isolated, cultured, and CO and NO release, ATP, cyclic-guanosyl monophosphate (cGMP), and lipid peroxide (LPO) content of cells, as well as phosphatidylcholine (PC)synthesis, were measured. Hepatocytes isolated from old ovariectomized rats showed a decrease in ATP content and PC synthesis compared to young rats. Age also induced an increase in LPO, NO, CO, and cGMP. Treating old rats with GH significantly increased ATP and reduced CO and cGMP levels. Estradiol administration improved all the parameters that were altered. Co-administration of GH and estrogens induced a more marked effect than estrogens alone only in cGMP content. In conclusion, administration of estrogens to old ovariectomized females seemed to prevent oxidative changes in hepatocytes, whereas the effect of GH is not so evident. PMID- 15805581 TI - Serum levels of leptin, insulin, and lipids in relation to breast cancer in china. AB - Epidemiological studies have found obesity to be a risk factor for women's breast cancer. The present study was to investigate whether there is a relationship between serum levels of leptin, insulin, and lipids and breast cancer incidence, in order to find experimental evidence that would be helpful in the diagnosis and prevention of breast cancer. Blood samples were collected from 130 patients with mammary disease and 103 healthy control subjects. Serum leptin, insulin, and lipids were determined by radioimmunoassay (RIA), enzyme-linked immunosorbent assays (ELISA), and Biochemistry Auto-analyzer, respectively. The data analysis was performed by use of the SPSS10.0 computer software. We found that the serum levels of leptin, insulin, and triglyceride (TG) were clearly higher in patients with breast cancer than in patients with benign breast disease and healthy controls, while serum HDL-C levels were lower in breast cancer patients (p < 0.03). Moreover, serum leptin levels were significantly correlated with BMI (body mass index) among three groups, whereas serum insulin levels were unrelated to BMI among three groups. Furthermore, the serum levels of leptin and insulin were not associated with menopausal status in patients with mammary disease (p > 0.05); however, the serum levels of F-Chol, T-Chol, TG, LDL-C, and APOB were significant higher in postmenopausal cases than those in premenopausal cases (p < 0.025). Interestingly, logistic regression analysis showed that subjects with elevated serum levels of leptin, insulin, TG, APOA1, and reduced level of serum HDL-C displayed increased risk of developing breast cancer than those with the normal levels, respectively. In conclusion, the present study suggested that aberrant serum levels of leptin, insulin, and lipids might play an important role in carcinogenesis of breast cancer. The elevated serum levels of leptin, insulin, TG, APOA1, and reduced level of serum HDL-C may be correlated with increased risk of breast cancer, suggesting that one way of preventing breast cancer would be carried out by controlling the intake of food. PMID- 15805583 TI - Ovulation block by Pueraria mirifica: a study of its endocrinological effect in female monkeys. AB - Pueraria mirifica (PM), a Thai herb containing phytoestrogens, may act as estrogen and disturb reproduction. To investigate the effect of PM on the menstrual cycle length and related hormones, nine adult female monkeys (Macaca fascicularis) were separated into three groups. Each group (n = 3) was fed with 10, 100, and 1000 mg/d of PM for three menstrual cycles. The menstrual cycle length increased significantly in monkeys treated with PM-10 and PM-100 and disappeared completely in monkeys treated with PM-1000. Serum follicle stimulating hormone, luteinizing hormone, estradiol, progesterone, and irinhibin were lower during the treatment period in a dose-dependent manner. Changes in menstrual cycle length and the hormonal levels recovered during the post treatment period only in monkeys treated with PM-10 and PM-100. PM greatly influences menstrual cycles and may suppress ovulation by lowering serum levels of gonadotropins. PMID- 15805582 TI - Retinoid-related receptor (ROR) alpha mRNA expression is altered in the brain of male mice lacking all ligand-binding thyroid hormone receptor (TR) isoforms. AB - In the vertebrate brain, the thalamus serves as a relay and integration station for diverse neuronal information en route from the periphery to the cortex. Deficiency of TH during development results in severe cerebral abnormalities similar to those seen in the mouse when the retinoic acid receptor (ROR)alpha gene is disrupted. To investigate the effect of the thyroid hormone receptors (TRs) on RORalpha gene expression, we used intact male mice, in which the genes encoding the alpha and beta TRs have been deleted. In situ hybridization for RORalpha mRNA revealed that this gene is expressed in specific areas of the brain including the thalamus, pons, cerebellum, cortex, and hippocampus. Our quantitative data showed differences in RORalpha mRNA expression in different subthalamic nuclei between wild-type and knock-out mice. For example, the centromedial nucleus of the thalamus, which plays a role in mediating nociceptive and visceral information from the brainstem to the basal ganglia and cortical regions, has less expression of RORalpha mRNA in the knockout mice (-37%) compared to the wild-type controls. Also, in the dorsal geniculate (+72%) and lateral posterior nuclei (+58%) we found more RORalpha mRNA in dKO as compared to dWT animals. Such differences in RORalpha mRNA expression may play a role in the behavioral alterations resulting from congenital hypothyroidism. PMID- 15805584 TI - Low level of cyclin D1 protein expression in thyroid microcarcinomas from an autopsy series. AB - In a recent epidemiological screening study in an autopsy series, we found a high prevalence of microcarcinomas (MCs) (21/443 = 4.74%). We found no iodine intake-, gender-, or age-dependent differences in the prevalence of MCs. The results suggest a different and benign behavior of MCs compared to clinical cancer. The role of cyclin D1 overexpression in the pathogenesis of thyroid tumors is not known clearly; however, overexpression of this protein was reported in well differentiated papillary cancers and in incidentally found metastasizing MCs. To date, cyclin D1 expression has not been investigated in autopsy-derived thyroid MCs. Eight MCs were available for immunostaining and comparison with 15 clinically detected papillary thyroid cancers. Fourteen out of 15 clinical carcinomas expressed cyclin D1 (93.3%), while in the MCs this ratio was 1 out of 8 (12.5%) (p = 0.0001). The only cyclin D1-positive MC was multifocal (both lobes of the gland were affected). We concluded that the benign behavior of most autopsy-derived MCs may be associated with the lack of cyclin D1 overexpression. PMID- 15805585 TI - Structure and functional analysis of unclassified genes strongly expressed in human visceral adipose tissue. AB - Our previous work has described the gene expression patterns of human visceral adipose tissue (VAT) at the transcriptome level and reported that the strongly expressed genes in VAT showed an uneven distribution throughout the genome. The aim of the present work was to focus on the unclassified genes and known expressed sequence tags (ESTs) strongly expressed in VAT and analyze their structure and function with bioinformatics. Among the 400 ESTs strongly expressed in the VAT, 340 clones were classified into known genes through searching the latest Genbank database. Functional classification showed that 85 clones were unclassified known genes, and approx 90% of them were found to be expressed in adipose tissue for the first time. Among the 85 unclassified genes, only two share similarities in the coding sequences with all species examined, and six genes had so far no obvious similarity to any genes across different species. The protein products of 7 genes had putative signal peptide and 11 had transmembrane domains. The protein products of 39 genes had relative specific motifs or prosites on primary structure. In silico Northern blot showed that 21 known ESTs were abundantly specifically expressed in adipose tissue, which may provide clues to identify novel genes closely related to adipocyte function with potential pathophysiological implications. PMID- 15805586 TI - Involvement of estrogen in the pathogenesis of cyclophosphamide-induced cystitis in rats. AB - We examined the effects of ovariectomy and castration on the histology of cyclophosphamide (CP)-induced cystitis in rats. The animals were injected with CP (100 mg/kg) or saline intraperitoneally twice with an intervening 4 d and were euthanized at 1 or 2 wk after the initial CP administration. Saline treatment did not cause an apparent histological change in the bladder regardless of surgery, i.e., ovariectomy, castration, and sham-operation. CP treatment resulted in submucosal edema, urothelial damage, hemorrhage, and leukocyte infiltration in the bladder at 1 wk after the initial CP administration regardless of surgery. However, these histological changes were more severe in ovariectomized rats than in the sham-operated rats. In contrast, there were no significant differences in CP-induced histological changes in the bladder between castrated and sham operated groups. We further examined the role of estrogen and progesterone on the CP-induced histology in the bladder by the replacement with estrogen only or estrogen plus progesterone for 2 wk in overiectomized rats. Estrogen treatment ameliorated CP-induced histological changes compared to oil treatment, whereas estrogen plus progesterone treatment did not produce any differences in the histology of the bladder compared to estrogen treatment. These results suggest that estrogen may play a role in the pathogenesis of bladder inflammation. PMID- 15805587 TI - Lack of increase of serum adiponectin concentrations with a moderate weight loss during six months on a high-caloric diet in military service among a young male Finnish population. AB - An increase of insulin resistance and a worsening of lipid profile during 6 mo of military service in young male Finnish population has previously been shown by us. The present study demonstrates unfavorable changes of serum adiponectin concentrations and their association with weight loss in these particular circumstances. Adiponectin in a range of 4.3-21.2 microg/mL was present in the serum samples and had a significant negative correlation with weight, body mass index, waist to hip ratio, and insulin. Fasting serum lipids and plasma insulin significantly increased and serum adiponectin levels significantly decreased during the military service. Even the subjects with a 5-10% decrease in body weight showed the same result. In cases with more than 10% weight reduction and a significant decrease of fasting insulin concentration, the total and low density lipoprotein cholesterol significantly increased and adiponectin concentration tended to decreased. Only in severely obese cases (BMI >/= 30 kg/m2) with more than 10% decrease in body mass index adiponectin levels tended to increase, although not statistically significantly. This study shows that serum adiponectin concentrations decreased during a 6 mo high-caloric diet in military service, and even a moderate weight reduction induced by high-energy expenditure in exercise during service did not increase its levels. PMID- 15805589 TI - Structure of pyrR (Rv1379) from Mycobacterium tuberculosis: a persistence gene and protein drug target. AB - The Mycobacterium tuberculosis pyrR gene (Rv1379) encodes a protein that regulates the expression of pyrimidine-nucleotide biosynthesis (pyr) genes in a UMP-dependent manner. Because pyrimidine biosynthesis is an essential step in the progression of TB, the gene product pyrR is an attractive antitubercular drug target. The 1.9 A native structure of Mtb pyrR determined by the TB Structural Genomics Consortium facilities in trigonal space group P3(1)21 is reported, with unit-cell parameters a = 66.64, c = 154.72 A at 120 K and two molecules in the asymmetric unit. The three-dimensional structure and residual uracil phosphoribosyltransferase activity point to a common PRTase ancestor for pyrR. However, while PRPP- and UMP-binding sites have been retained in Mtb pyrR, a distinct dimer interaction among subunits creates a deep positively charged cleft capable of binding pyr mRNA. In silico screening of pyrimidine-nucleoside analogs has revealed a number of potential lead compounds that, if bound to Mtb pyrR, could facilitate transcriptional attenuation, particularly cyclopentenyl nucleosides. PMID- 15805590 TI - Structure of CDP-D-glucose 4,6-dehydratase from Salmonella typhi complexed with CDP-D-xylose. AB - Tyvelose is a unique 3,6-dideoxyhexose found in the O antigens of some pathogenic species of Yersinia and Salmonella. It is produced via a complex biochemical pathway that employs CDP-D-glucose as the starting ligand. CDP-D-glucose 4,6 dehydratase catalyzes the first irreversible step in the synthesis of this 3,6 dideoxysugar by converting CDP-D-glucose to CDP-4-keto-6-deoxyglucose via an NAD+ -dependent intramolecular oxidation-reduction reaction. Here, the cloning, protein purification and X-ray crystallographic analysis of CDP-D-glucose 4,6 dehydratase from Salmonella typhi complexed with the substrate analog CDP-D xylose are described. Each subunit of the tetrameric enzyme folds into two domains. The N-terminal region contains a Rossmann fold and provides the platform for NAD(H) binding. The C-terminal motif is primarily composed of alpha-helices and houses the binding pocket for the CDP portion of the CDP-D-xylose ligand. The xylose moiety extends into the active-site cleft that is located between the two domains. Key residues involved in anchoring the sugar group to the protein include Ser134, Tyr159, Asn197 and Arg208. Strikingly, Ser134 O gamma and Tyr159 O eta sit within 2.9 A of the 4'-hydroxyl group of xylose. Additionally, the side chains of Asp135 and Lys136 are located at 3.5 and 3.2 A, respectively, from C-5 of xylose. In the structurally related dTDP-D-glucose 4,6-dehydratase, the Asp/Lys pair is replaced with an Asp/Glu couple. On the basis of this investigation, it can be speculated that Tyr159 serves as the catalytic base to abstract the 4'-hydroxyl proton from the sugar and that Asp135 and Lys136 play critical roles in the subsequent dehydration step that leads to the final product. PMID- 15805588 TI - Further evidence for the regulation of acetyl-CoA carboxylase activity by a glutamate- and magnesium-activated protein phosphatase in the pancreatic beta cell: defective regulation in the diabetic GK rat islet. AB - We previously identified and characterized a glutamate- and magnesium-sensitive PP2A-like phosphatase (GAPP), which dephosphorylated and activated acetyl-CoA carboxylase (ACC) in the islet beta cell. Herein, we studied potential regulatory mechanisms by which GAPP is activated by glutamate and magnesium, and also quantitated the degree of activation, by glutamate- and magnesium, of ACC in normal rat islets and islets derived from the diabetic Goto-Kakizaki (GK) rat, a model for type 2 diabetes in humans. Our findings indicate that magnesium, but not glutamate, specifically activates the post-translational carboxylmethylation (CML) of the 36 kDa catalytic subunit of GAPP. Okadaic acid (OKA), which inhibits GAPP-mediated activation of ACC, also reduced the magnesium-stimulated CML of the catalytic subunit of GAPP in all the beta cell preparations studied. These data suggest that the CML step may be necessary for magnesium- and glutamate-mediated activation of ACC. We also observed a marked attenuation in magnesium- and glutamate-facilitated activation of ACC activity in islets derived from the GK rat. Together, our findings raise an interesting possibility that inhibition of GAPP-catalyzed inactivation of ACC (and subsequent reduction in the generation of long-chain fatty acids) could contribute toward the abnormalities in insulin secretion demonstrable in this animal model for type 2 diabetes. PMID- 15805591 TI - Structure of zinc-independent sorbitol dehydrogenase from Rhodobacter sphaeroides at 2.4 A resolution. AB - Recombinant sorbitol dehydrogenase (SDH) from Rhodobacter sphaeroides has been crystallized in the absence of the cofactor NAD(H) and its structure determined to 2.4 A resolution using molecular replacement (refined R and R free factors of 18.8 and 23.8%, respectively). As expected from the sequence and shown by the conserved fold, SDH can be assigned to the short-chain dehydrogenase/reductase protein family. The cofactor NAD and the substrate sorbitol have been modelled into the structure and the active-site architecture, which displays the highly conserved catalytic tetrad of Asn-Ser-Tyr-Lys residues, is discussed in relation to the enzyme mechanism. This is the first structure of a bacterial SDH belonging to the SDR family. PMID- 15805592 TI - Crystallization of mitochondrial respiratory complex II from chicken heart: a membrane-protein complex diffracting to 2.0 A. AB - A procedure is presented for preparation of diffraction-quality crystals of a vertebrate mitochondrial respiratory complex II. The crystals have the potential to diffract to at least 2.0 A with optimization of post-crystal-growth treatment and cryoprotection. This should allow determination of the structure of this important and medically relevant membrane-protein complex at near-atomic resolution and provide great detail of the mode of binding of substrates and inhibitors at the two substrate-binding sites. PMID- 15805593 TI - Automatic structure determination based on the single-wavelength anomalous diffraction technique away from an absorption edge. AB - The phasing of macromolecular structures based on the use of the single wavelength anomalous diffraction method has recently enjoyed a revival. Here, additional evidence is provided that the method may be successfully applied at wavelengths remote from the absorption edge of interest and that it is in principle applicable to a large number of systems. This opens up the possibility of rapid and reliable automatic de novo structure determination using simple experimental configurations with no need for wavelength tunability or absorption edge scanning. The method should therefore be exploitable at most synchrotron beamlines. The effects of data completeness and multiplicity on the quality of the phases obtained are discussed as are the prospects for the automation of macromolecular structure solution based on the experimental protocols described. PMID- 15805594 TI - Structure of a liganded type 2 non-specific lipid-transfer protein from wheat and the molecular basis of lipid binding. AB - In plants, a family of ubiquitous proteins named non-specific lipid-transfer proteins (ns-LTPs) facilitates the transfer of fatty acids, phospholipids and steroids between membranes. Recent data suggest that these secreted proteins play a key role in the formation of cuticular wax layers and in defence mechanisms against pathogens. In this study, X-ray crystallography has been used to examine the structural details of the interaction between a wheat type 2 ns-LTP and a lipid, L-alpha-palmitoyl-phosphatidyl glycerol. This crystal structure was solved ab initio at 1.12 A resolution by direct methods. The typical alpha-helical bundle fold of this protein is maintained by four disulfide bridges and delineates two hydrophobic cavities. The inner surface of the main cavity is lined by non-polar residues that provide a hydrophobic environment for the palmitoyl moiety of the lipid. The head-group region of this lipid protrudes from the surface and makes several polar interactions with a conserved patch of basic residues at the entrance of the pocket. The alkyl chain of a second lipid is bound within an adjacent smaller cavity. The structure shows that binding of the lipid tails to the protein involves extensive hydrophobic interactions. PMID- 15805595 TI - Crystal and solution structures of 7-amino-actinomycin D complexes with d(TTAGBrUT), d(TTAGTT) and d(TTTAGTTT). AB - The formation of the complex of 7-amino-actinomycin D with potentially single stranded DNA has been studied by X-ray crystallography in the solid state, by NMR in solution and by molecular modelling. The crystal structures of the complex with 5'-TTAG[Br(5)U]T-3' provide interesting examples of MAD phasing in which the dispersive component of the MAD signal was almost certainly enhanced by radiation damage. The trigonal and orthorhombic crystal modifications both contain antibiotic molecules and DNA strands in the form of a 2:4 complex: in the orthorhombic form there is one such complex in the asymmetric unit, while in the trigonal structure there are four. In both structures the phenoxazone ring of the first drug intercalates between a BrU-G (analogous to T-G) wobble pair and a G-T pair where the T is part of a symmetry-related molecule. The chromophore of the second actinomycin intercalates between the BrU-G and G-BrU wobble pairs of the partially paired third and fourth strands. The base stacking also involves (A*T)*T triplets and Watson-Crick A-T pairs and leads to similar complex three dimensional networks in both structures, with looping-out of unpaired bases. Although the available NOE constraints of a solution containing the antibiotic and d(TTTAGTTT) strands in the ratio 1:1 are insufficient to determine the structure of the complex from the NMR data alone, they are consistent with the intercalation geometry observed in the crystal structure. Molecular-dynamics (MD) trajectories starting from the 1:2 complexes observed in the crystal showed that although the thymines flanking the d(AGT) core are rather flexible and the G-T pairing is not permanently preserved, both strands remain bound to the actinomycin by strong interactions between it and the guanines between which it is sandwiched. Similar strong binding (hemi-intercalation) of the actinomycin to a single guanine was observed in the MD trajectories of a 1:1 complex. The dominant interaction is between the antibiotic and guanine, but the complexes are stabilized further by promiscuous base-pairing. PMID- 15805596 TI - Combined use of AFM and X-ray diffraction to analyze crystals of an engineered, domain-deleted antibody. AB - A genetically engineered humanized C(H)2-domain-deleted monoclonal antibody lacking any interchain-hinge disulfide bonds has been crystallized in the presence of detergent in a form suitable for X-ray diffraction analysis. The crystals were grown from 4 M formate along with Triton X-100 and had P2(1)2(1)2 space-group symmetry, with unit-cell parameters a = 83, b = 224, c = 167 A. The crystals diffract to beyond 2.8 A resolution. A disordered crystal form of larger size and more attractive habit was also grown from 4 M formate, but in the presence of the Anapoe series of detergents. Preliminary X-ray data, in conjunction with atomic force microscopy images, are consistent with asymmetric units consisting of two intact antibodies forming a circular dimeric ring. The crystallizing unit, which must contain a twofold axis, is a toroidal assembly of four antibodies (two dimeric rings). Competition between dimers and tetramers to enter the lattice, along with a unique kind of planar defect of packing, may be responsible for the unusually high defect density and the disorder of the X-ray diffraction pattern exhibited by the second crystal form. An approach to crystallizing proteins showing phase separation, particularly intact antibodies, that uses a preliminary detergent test set is described. PMID- 15805597 TI - Synchrotron X-ray powder diffraction study of hexagonal turkey egg-white lysozyme. AB - The structure of turkey egg-white lysozyme (TEWL) has been refined from high resolution X-ray powder diffraction data. The sample was rapidly obtained as a polycrystalline precipitate at high protein concentration using 0.5 M NaCl solvent pH 6 and was deposited in the PDB with code 1xft. The diffraction data were collected at room temperature. Molecular replacement was shown to give a suitable starting point for refinement, illustrating that powder data can be sufficient for this approach. Crystallographic models were then refined by combined Rietveld and stereochemical restraint analysis of the powder data (d(min) = 3.35 A), resulting in the extraction of reliable lattice parameters and the refinement of the molecular conformation at room temperature. The structure is hexagonal [space group P6(1)22, unit-cell parameters a = 71.0862 (3), c = 85.0276 (5) A] with 12 symmetry-related molecules in the unit cell, in agreement with previous studies. The results of our analysis are indicative of specific amino acids being disordered at this temperature. Upon cooling, a sudden drop in the lattice parameters at approximately 250 K is observed concurrently with the freezing of the mother liquor. The observation of severe peak broadening below this temperature indicates strain effects accompanying the freezing transition, which are found to be reversible. Finally, a correlation between the unit-cell parameters and the pH of the buffer solution is evident, in a similar manner to earlier observations on HEWL. PMID- 15805598 TI - Structure of gramicidin D-RbCl complex at atomic resolution from low-temperature synchrotron data: interactions of double-stranded gramicidin channel contents and cations with channel wall. AB - Gramicidin D (gD) is a naturally occurring ionophoric antibiotic that forms membrane channels specific for monovalent cations. The crystal structure of the RbCl complex of gD has been determined at 1.14 A resolution from low-temperature (100 K) synchrotron-radiation data with a final R of 16%. The structure was refined with anisotropic temperature factors for all non-H atoms and with partial occupancies for many of them. The asymmetric unit in the crystal contains four crystallographically independent molecules that form two right-handed antiparallel double-stranded dimers. There are seven distinct rubidium-binding sites in each dimeric channel. The occupancy factors of Rb cations are between 0.11 and 0.35 and the total ion contents of the two crystallographically independent channels are 1.59 and 1.22 ions, respectively. Although each channel is 'chemically symmetrical', the side-chain conformations, the distributions of rubidium cations and their binding sites in the two independent channels are not. Cations are 'coordinated' by delocalized pi-electrons of three to five carbonyl groups that together with peptide backbone chains form the gramicidin channel walls. The water:cation ratio in the channel interior is four or five:one, and five or six waters separate Rb cations during their passage through the channel. PMID- 15805599 TI - Structures of complexes between echinomycin and duplex DNA. AB - The structure of the bis-intercalation complex of the depsipeptide antibiotic echinomycin with (CGTACG)2 has been redetermined at a higher resolution (1.4 A) and new high-resolution structures (1.1-1.5 A) are reported for the complexes of echinomycin with (GCGTACGC)2 (at both low and high ionic strengths) and (ACGTACGT)2. The structures show the expected Hoogsteen pairing for the base pairs flanking the intercalating chromophores on the outside and Watson-Crick pairing for both base pairs enclosed by the echinomycin. In the octamer complexes but not the hexamer complex, the echinomycin molecule, which would possess a molecular twofold axis were it not for the thioacetal bridge, shows twofold disorder. In all the structures the stacking of the base pairs and chromophores is extended by intermolecular stacking. The structures provide more precise details of the hydrogen bonding and other interactions between the bis intercalating antibiotics and the duplex DNA than were previously available. PMID- 15805600 TI - Auto-rickshaw: an automated crystal structure determination platform as an efficient tool for the validation of an X-ray diffraction experiment. AB - The EMBL-Hamburg Automated Crystal Structure Determination Platform is a system that combines a number of existing macromolecular crystallographic computer programs and several decision-makers into a software pipeline for automated and efficient crystal structure determination. The pipeline can be invoked as soon as X-ray data from derivatized protein crystals have been collected and processed. It is controlled by a web-based graphical user interface for data and parameter input, and for monitoring the progress of structure determination. A large number of possible structure-solution paths are encoded in the system and the optimal path is selected by the decision-makers as the structure solution evolves. The processes have been optimized for speed so that the pipeline can be used effectively for validating the X-ray experiment at a synchrotron beamline. PMID- 15805601 TI - Likelihood-enhanced fast translation functions. AB - This paper is a companion to a recent paper on fast rotation functions [Storoni et al. (2004), Acta Cryst. D60, 432-438], which showed how a Taylor-series expansion of the maximum-likelihood rotation function leads to improved likelihood-enhanced fast rotation functions. In a similar manner, it is shown here how linear and quadratic Taylor-series expansions and least-squares approximations of the maximum-likelihood translation function lead to likelihood enhanced translation functions, which can be calculated by FFT and which are more sensitive to the correct translation than the traditional correlation-coefficient fast translation function. These likelihood-enhanced translation targets for molecular-replacement searches have been implemented in the program Phaser using the Computational Crystallography Toolbox (cctbx). PMID- 15805602 TI - Structure of a pseudomerohedrally twinned monoclinic crystal form of a pyridoxal phosphate-dependent catalytic antibody. AB - The purification, crystallization and structure determination at 2.3 A resolution of the complex of the pyridoxal-5'-phosphate (PLP) dependent catalytic antibody 15A9 with a phosphopyridoxyl-L-alanine (PPL-L-alanine) substrate analogue are described. The crystal belongs to space group P2(1), with two molecules in the asymmetric unit related by non-crystallographic symmetry. The unit-cell parameters are a = 63.5, b = 81.7, c = 79.3 A and beta is fortuitously 90 degrees . Refinement of the structure converged at unacceptably high R factors. Although the traditional analysis of intensity distribution did not indicate twinning, pseudomerohedral twinning was revealed by a newer test based on local intensity differences [Padilla & Yeates (2003), Acta Cryst. D59, 1124-1130]. When the potential twinning operator was included in SHELX, the structure could be satisfactorily refined with a twinning fraction of 0.46, indicating a nearly perfect hemihedrally twinned crystal. One of the active sites is occupied by the phosphopyridoxyl-L-alanine ligand, while one iodide ion mimics the cofactor phosphate group in the other. Four other iodide ions are present in the structure: two are involved in specific intermolecular contacts and two dictate the conformation of the CDRH3 loop in each molecule. PMID- 15805603 TI - Favourable mediation of crystal contacts by cocoamidopropylbetaine (CAPB). AB - Crystals of excellent quality are a prerequisite for high-resolution X-ray data. However, in refinement protocols of crystallization conditions it is often difficult to obtain the right combination of, for example, protein concentration, drop size, temperature and additives. A novel approach for optimizing crystal contacts in a most favourable fashion by performing crystallization setups with the zwitterionic surfactant cocoamidoproylbetaine (CAPB) is introduced. In the presence of this surfactant, highly diffracting crystals were obtained. Here, data from a right-handed coiled coil (RHCC) in complex with CAPB at 1.4 A resolution are presented. The addition of CAPB using otherwise identical crystallization conditions and the same X-ray source caused an improvement in resolution from 2.9 to 1.4 A. PMID- 15805604 TI - Structure of S35C flavodoxin mutant from Desulfovibrio vulgaris in the semiquinone state. AB - The crystallographic structure of an engineered flavodoxin mutant from Desulfovibrio vulgaris has been analysed. Site-directed mutagenesis was used to substitute serine 35 with a cysteine to provide a possible covalent linkage. The crystal structure of the semiquinone form of this mutant is similar to the corresponding oxidation state of the wild-type flavodoxin. Analysis of the structural changes reveals the interaction between N(5)H of the flavin and the carbonyl O atom of Gly61 to be critical for modulation of the electrochemical properties of the protein. PMID- 15805605 TI - Unusual non-crystallographic symmetry in crystals of a 420 kDa crustacean clottable protein. AB - The 420 kDa hemolymph clottable protein has been purified from the California spiny lobster Panulirus interrruptus and crystallized in both monoclinic and orthorhombic space groups. Complete data sets have been collected from the two crystal forms to a maximum resolution of 3.9 and 3.2 A, respectively. The monoclinic crystals exhibit unusual noncrystallographic symmetry with pseudo ninefold rotational symmetry generated from three dimers per asymmetric unit. The orthorhombic crystals have a single dimer per asymmetric unit. Attempts to phase the data are currently under way. PMID- 15805606 TI - Expanding screening space through the use of alternative reservoirs in vapor diffusion experiments. AB - Setting up vapor-diffusion crystallization experiments against four different reservoir solutions showed that the reservoir solution may have a profound effect on the outcome of a crystallization experiment. This suggests that a facile way to increase crystallization space through screening is not to add more crystallization conditions to the process, but to set up the same conditions over different reservoirs. PMID- 15805607 TI - Thallium selenate (Tl2SeO4) in a paraelastic phase by X-ray powder diffraction. AB - The structure of thallium selenate, Tl2SeO4, in a paraelastic phase (above 661 K) has been analysed by Rietveld analysis of the X-ray powder diffraction pattern. Atomic parameters based on the isomorphic K2SO4 crystal in the paraelastic phase were used as the starting model. The structure was determined in the hexagonal space group P6(3)/mmc, with a = 6.2916 (2) A and c = 8.1964 (2) A. From the Rietveld refinement it was found that two orientations are possible for the SeO4 tetrahedra, in which one of their apices points randomly up and down with respect to [001]. One Tl atom lies at the origin with 3m symmetry, the other Tl and one of the O atoms occupy sites with 3m symmetry, the Se atom is at a site with 6m2 symmetry and the remaining O atom is at a site with m symmetry. Furthermore, it was also found that the Tl atoms display anomalously large positional disorder along [001] in the paraelastic phase. PMID- 15805608 TI - catena-Poly[[aquabis(1H-benzimidazole-kappa N3)manganese(II)]-mu-adipato]. AB - In the title polymeric complex, [Mn(C6H8O4)(C7H6N2)2(H2O)]n, the MnII atom is surrounded by two adipate dianions, two benzimidazole molecules and one coordinated water molecule. The Mn atoms and coordinated water molecule are located on a twofold axis, and the bridging adipate ligand is located on an inversion center. The adipate dianions bridge neighboring MnII atoms to form polymeric chains. Each MnII atom is seven-coordinate, the longest Mn-O bond length being 2.5356 (16) A. PMID- 15805609 TI - Coordination polymers formed by bridging 2-substituted tetrazole ligands: poly[[dichlorocopper(II)]-di-mu2-2-propyl-2H-tetrazole-kappa2N1:N4] and poly[[dichlorocopper(II)]-di-mu2-2-allyl-2H-tetrazole-kappa2N1:N4]. AB - Two polymeric complexes, [CuCl2L2]n, where L is 2-propyltetrazole (C4H8N4) or 2 allyltetrazole (C4H6N4), are the first coordination polymers of 2-substituted tetrazoles in which only the tetrazole rings bridge neighbouring Cu atoms. In both complexes, the Cu atoms lie on inversion centres and are six-coordinated in tetragonally distorted octahedral geometries, CuCl2N4, with two N1 tetrazole ring atoms in the axial positions and two Cl atoms and two N4 tetrazole ring atoms in the equatorial sites. The Cl atoms do not participate in the polymeric layer formation. PMID- 15805610 TI - Layered poly[mu2-aqua-mu3-dimethylformamide-di-mu3-phenylphosphonato dipotassium]. AB - The first structurally characterized alkali metal phosphonate, the title compound, [K2(C6H6O3P)2(C3H7NO)(H2O)]n, has a complex structure, with layers parallel to the crystallographic bc plane consisting of two crystallographically independent K atoms sandwiched between the three types of ligands present in the structure, viz. water molecules, dimethylformamide molecules and two crystallographically independent phenylphosphonate ligands. Six O atoms coordinate to one K atom and seven to the other. The interlayer distance is 15.327 (4) A. The K-O distances are in the range 2.739 (2)-2.932 (2) A for the seven-coordinate K atom and 2.650 (2)-2.821 (2) A for the six-coordinate K atom. PMID- 15805611 TI - Poly[nickel(II)-di-mu-4,4'-bipyridyl-kappa4N:N'-mu-dichromato-kappa2O:O'] and poly[copper(II)-di-mu-4,4'-bipyridyl-kappa4N:N'-mu-dichromato-kappa2O:O']. AB - The novel title hybrid isomorphous organic-inorganic mixed-metal dichromates, [Ni(Cr2O7)(C10H8N2)2] and [Cu(Cr2O7)(C10H8N2)2], have been synthesized. A non centrosymmetric three-dimensional (4,6)-net is formed from a linear chain of vertex-linked [Cr2O7]2- and [MN4O2]2+ (M = Ni and Cu) units, which in turn are linked by the planar bidentate 4,4'-bipyridine ligand through the four remaining vertices of the [MN4O2]2+ octahedra. There are two such three-dimensional nets that interpenetrate with inversion symmetry. PMID- 15805612 TI - Bis(di-2-pyridyl sulfide-kappa2N,N')(4-methylpyrimidin-2-yl 2-pyridylmethyl sulfide-kappa2N,S)ruthenium(II) bis(hexafluorophosphate) acetonitrile solvate. AB - The crystal structure of the title compound, [Ru(C10H8N2S)2(C11H11N3S)](PF6)2.C2H3N, is composed of a bivalent octahedral RuII complex, two PF6- anions and an acetonitrile solvent molecule. Two PF6- units are found on a crystallographic binary axis, therefore contributing just one half each to the asymmetric unit cell. The structure displays a peculiar stereochemistry of the cation. Three bidentate ligands around the Ru centre, together with the coordination of the non-symmetric S atom, mean that these two atoms are chiral. This would lead to four stereoisomers, but only an enantiomeric pair was found in the analyzed sample. PMID- 15805613 TI - Bis(mu-pyridine-2-thiolato)-kappa4S:N;N:S-bis[(pyridine-2-thiolato-kappaS)(2 pyridylphenyl-kappa2N,C2)platinum(III)](Pt-Pt): the non-solvated and acetonitrile solvated forms. AB - The title dinuclear platinum(III) complex, [Pt2(C11H8N)2(C5H4NS)4], forms two crystal structures, viz. the non-solvated and acetonitrile-solvated (C2H3N) forms. For both forms, two (2-pyridylphenyl)platinum units are bridged by two pyridine-2-thiolate (pyt) anions in a head-to-tail configuration, and the other two pyridine-2-thiolate anions occupy the axial position, coordinated through their S atoms. The most remarkable difference between the two forms is the orientation of the axial monodentate ligands. Those for the solvated form are located over the 2-pyridylphenyl ligands, being related by a twofold axis which lies through the centre of the Pt-Pt bond, while the axial pyt ligands for the non-solvated form are oriented irregularly, which is attributable to the dimeric arrangement in the crystal. PMID- 15805614 TI - A mixed-valence CuII/CuI anion-cation complex: bis[mu-5-sulfosalicylato(3 )]bis[(di-2-pyridylamine)copper(II)] bis[bis(di-2-pyridylamine)copper(I)] dihydrate. AB - The title compound, [CuII(2)(C7H3O6S)2(C10H9N3)2][CuI(C10H9N3)2]2.2H2O, consists of anionic CuII moieties, cationic CuI species and uncoordinated water molecules. The anionic dimeric unit consists of one crystallographically independent fully deprotonated 5-sulfosalicylate (2-oxido-5-sulfonatobenzoate) anion, a di-2 pyridylamine group and a CuII atom. Each CuII atom is five-coordinate within a square-pyramidal geometry. The anion lies on a special position of -1 site symmetry. In the cationic monomer, the CuI atom adopts tetrahedral geometry. The cations and anions are connected by O-H...O and N-H...O hydrogen bonds. PMID- 15805615 TI - Pseudo-merohedrally twinned tetrakis(1H-imidazole-kappaN3)bis(N-nitrocyanamidato kappaN)copper(II). AB - Crystals of the title complex, [Cu(CN3O2)2(C3H4N2)4], the structure of which has been determined by single-crystal X-ray diffraction at 304 K, appear to be pseudo merohedrally twinned. Transformation to a monoclinic C-centred cell was necessary in order to derive the twin law. Twin refinement in a triclinic unit cell significantly reduced the R value. The asymmetric unit of the triclinic cell consists of one molecule in a general position and two half entities with the Cu atom on a centre of inversion. The coordination of the Cu atom is quasi octahedral, with four imidazole N-atom donors in the equatorial plane and two cyano N atoms from the N-nitrocyanamidate anion in axial positions. Owing to symmetry in the centrosymmetric molecules, the trans imidazole ligands are parallel, while those in the non-centrosymmetric molecule make angles of 22.8 (2) and 77.9 (2) degrees . PMID- 15805616 TI - trans-Bis(2,2-diphenylethylamine-kappaN)bis(5,5-diphenylhydantoinato kappaN3)copper(II) and its chloroform disolvate. AB - The crystal structures of the title compounds, [Cu(C15H11N2O2)2(C14H15N)2] and [Cu(C15H11N2O2)2(C14H15N)2].2CHCl3, respectively, have been determined. The red disolvate complex affords a square-planar CuN4 coordination environment in which the CuII atom lies on a centre of symmetry. The blue solvent-free complex affords a distorted square-pyramidal CuN4O coordination environment and adjacent molecules form centrosymmetric dimers. A comparison of the different crystal structures focuses on the role of the solvent molecules in supramolecular assemblies of the copper(II) complexes. PMID- 15805617 TI - Concerted coordination and hydrogen-bonding network of the 1,4-di-4-pyridyl-2,3 diaza-1,3-butadiene [Zn(MeOH)2(H2O)2](ClO4)2 3:1 complex. AB - In the title compound, diaquabis(1,4-di-4-pyridyl-2,3-diaza-1,3 butadiene)dimethanolzinc(II) bis(perchlorate) 1,4-di-4-pyridyl-2,3-diaza-1,3 butadiene methanol 1.72-solvate 1.28-hydrate, [Zn(C12H10N4)2(CH4O)2(H2O)2](ClO4)2.C12H10N4.1.72CH4O.1.28H2O, determined at ca 110 K, the Zn cation and the extended dipyridyl ligand both lie across inversion centres in space group P-1. The structure consists of a network arrangement of the constituent species stabilized by a combination of coordination, hydrogen bonding and pi-pi forces. Uncoordinated methanol and water solvent molecules occupy the otherwise void spaces within and between the networks. PMID- 15805619 TI - (mu-2,11-Dithia[3.3]paracyclophane-kappa2S:S')bis[(6-carboxypyridine-2 carboxylato-kappa2N,O6silver(I)]: a mixed-ligand silver-based dinuclear compound. AB - In the title complex, [Ag2(C7H4NO4)2(C16H16S2)], each Ag(I) atom is trigonally coordinated by one S atom of a 2,11-dithia[3.3]paracyclophane (dtpcp) ligand, and by one N and one O atom of a 6-carboxypyridine-2-carboxylate ligand. Dtpcp acts as a bidentate ligand, bridging two inversion-related AgI atoms to give a dinuclear silver(I) compound. The dinuclear moieties are interconnected via O H...O hydrogen bonds to form a two-dimensional zigzag sheet. Two such sheets are interwoven via pi-pi interactions between pyridine rings, affording an interwoven bilayer network. PMID- 15805618 TI - [(S)-1-Carbamoylethyl]bis(dimethylglyoximato-kappa2N,N')[(S)-1 phenylethylamine]cobalt(III) and bis(dimethylglyoximato-kappa2N,N')[(R)-1-(N methylcarbamoyl)ethyl][(R)-1-phenylethylamine]cobalt(III) monohydrate. AB - The title complexes, [Co(C3H6NO)(C4H7N2O2)2(C8H11N)] and [Co(C4H8NO)(C4H7N2O2)2(C8H11N)].H2O, were resolved from [(RS)-1 carbamoylethyl]bis(dimethylglyoximato)[(S)-1-phenylethylamine]cobalt(III) and bis(dimethylglyoximato)[(RS)-1-(N-methylcarbamoyl)ethyl][(R)-1 phenylethylamine]cobalt(III), respectively, and their crystal structures were determined in order to reveal the absolute configuration of the major enantiomer produced in the photoisomerization of each series of 2-carbamoylethyl and 2-(N methylcarbamoyl)ethyl cobaloxime complexes. PMID- 15805620 TI - 8-Aza-7-deaza-7-propynyladenosine methanol solvate. AB - In the title compound, 4-amino-3-propynyl-1-(beta-D-ribofuranosyl)-1H pyrazolo[3,4-d]pyrimidine methanol solvate, C13H15N5O4.CH3OH, the torsion angle of the N-glycosylic bond is between anti and high-anti [chi = -101.8 (5) degrees ]. The ribofuranose moiety adopts the C3'-endo (3T2) sugar conformation (N-type) and the conformation at the exocyclic C-C bond is +sc (gauche, gauche). The propynyl group is out of the plane of the nucleobase and is bent. The compound forms a three-dimensional network which is stabilized by several hydrogen bonds (O-H...O and O-H...N). The nucleobases are stacked head-to-tail. The methanol solvent molecule forms hydrogen bonds with both the nucleobase and the sugar moiety. PMID- 15805621 TI - 2-Amino-5-methylpyridinium (2-amino-5-methylpyridine)trichlorozincate(II). AB - The title compound, (C6H9N2)[ZnCl3(C6H8N2)], consists of one 2-amino-5 methylpyridinium cation and one (2-amino-5-methylpyridine)trichlorozincate(II) anion, which are held together by N-H...Cl hydrogen bonds and pi-pi interactions. The cation and the pyridine ligand show similar geometric features, except for the N-C bond lengths. Molecules of the title compound are connected by N-H...Cl hydrogen bonds to form chiral chains; these chains are associated further by C H...Cl hydrogen bonds to form layers, which are in turn linked by pi-pi interactions. PMID- 15805622 TI - L-valyl-L-serine trihydrate. AB - The valine side chains in the crystal structure of the title compound [systematic name: 2-(2-ammonio-3-methylbutanamido)-3-hydroxypropanoate trihydrate], C8H16N2O4.3H2O, stack along an a axis of 4.77 A to form hydrophobic columns surrounded by remarkable water/hydroxyl shells. The peptide main chains are connected by hydrogen bonds in two-dimensional layers. The peptide molecules in each layer are related only by translation, and generate a very rare pattern. This is rendered possible through the formation of the shortest C(alpha) H...O(carboxylate) interaction ever recorded. PMID- 15805623 TI - mer-Triiodotripyridineindium(III). AB - Crystals of the title compound, [InI3(C5H5N)3], consist of discrete molecules lying on a twofold axis running parallel to the crystallographic b axis. The molecules exhibit meridional octahedral stereochemistry, with In-I bond lengths of 2.8390 (6) and 2.8676 (3) A, and In-N bond lengths of 2.323 (5) and 2.309 (4) A. PMID- 15805624 TI - 21-deoxycortisone (17alpha-hydroxy-4-pregnene-3,11,20-trione). AB - The title compound, C21H28O4, a synthetic glucocorticoid, crystallizes with a single molecule in the asymmetric unit. Ring A is almost in a half-chair conformation, rings B and C are almost in chair conformations, and ring D is between a twist and a 13beta-envelope conformation. The A/B ring junction is quasi-trans, whereas the B/C and C/D ring junctions both approach trans characteristics. The molecule as a whole is slightly convex towards the beta side, with an angle of 9.60 (2) degrees between the C10-C19 and C13-C18 vectors. Molecular-packing and hydrogen-bonding (both intra- and intermolecular) interactions play a major role in the structural association of the compound. PMID- 15805625 TI - Bis[2-(3-pyridinio)benzimidazolium] di-mu-chloro-bis[trichlorocadmium(II)]. AB - The title compound, (C12H11N3)2[Cd2Cl8], consists of two discrete 2-(3 pyridinio)benzimidazolium cations and one [Cd2Cl8]4- anion. The dimeric [Cd2Cl8]4 anion lies about an inversion centre and consists of two distorted [CdCl5] trigonal bipyramids which share a common edge. The two Cd atoms are each coordinated by two mu-Cl atoms and three terminal Cl atoms, with a Cd...Cd separation of 3.9853 (6) A. The packing displays two-dimensional hydrogen-bonded sheets, which are further linked by C-H...Cl contacts and pi-pi stacking interactions to yield a three-dimensional network. PMID- 15805626 TI - Supramolecular structures of three isomeric 2-chloro-N (nitrophenyl)nicotinamides. AB - Molecules of 2-chloro-N-(2-nitrophenyl)nicotinamide, C12H8ClN3O3, are linked by two C-H...O hydrogen bonds into a chain of edge-fused R(2)(2) (14) and (4)(4) (24) rings. In 2-chloro-N-(3-nitrophenyl)nicotinamide monohydrate, C12H8ClN3O3.H2O, the molecules are linked by a combination of N-H...O, O-H...O and O-H...N hydrogen bonds into a chain of edge-fused rings containing two distinct types of R(4)(4) (16) ring. In 2-chloro-N-(4-nitrophenyl)nicotinamide, C12H8ClN3O3, which crystallizes with Z' = 2 in space group P2(1)/n, the molecules are linked by two N-H...N hydrogen bonds into simple C(2)(2) (12) chains. PMID- 15805627 TI - Poly[di-mu2-methanolato-di-mu2-tetramethoxyborato-disodium(I)]. AB - In the polymeric title compound, [Na2(C4H12BO4)2(CH4O)2]n, the two independent sodium cations are bound by five O atoms. All the O atoms of one tetramethoxyborate anion bind cations, forming a tetrameric cluster around a tetragonal inversion centre [Na-O = 2.2777 (18)-2.3907 (16) A]. Two methanol O atoms bridge the two Na atoms [Na-O = 2.3590 (15)-2.4088 (18) A] and provide the hydrogen-bonding H atoms. The second tetramethoxyborate anion provides two O atoms to one Na atom [mean Na-O = 2.31 (2) A] and two O atoms as donors for crosslinking hydrogen bonds to adjacent tetramers, which complete the three dimensional packing. The crystal was a treated as a racemic twin. PMID- 15805628 TI - Chains of edge-fused hydrogen-bonded R3 3(12) rings in N-phenyl-4 nitrophthalimide. AB - Molecules of the title compound [systematic name: 5-nitro-1H-isoindole-1,3(2H) dione], C14H8N2O4, adopt a conformation in the solid state which renders them chiral, and they are linked by three distinct types of direction-specific intermolecular interaction. The molecules are linked by two C-H...O hydrogen bonds [H...O = 2.50 and 2.52 A, C...O = 3.118 (7) and 3.294 (7) A, and C-H...O = 123 and 139 degrees ] into chains of edge-fused R(3)(3) (12) rings, which are themselves weakly linked into sheets by a combination of an aromatic pi-pi stacking interaction and a sheared-parallel carbonyl-carbonyl interaction. PMID- 15805629 TI - (eta5-cyclopentadienyl)(N,N-dimethyldithiocarbamato-kappa2S,S')[eta4 tetrakis(trifluoromethyl)cyclobutadienyl]molybdenum(IV). AB - The title complex, [Mo(C8F12)(C5H5)(C3H6NS2)], contains both a eta4-C4(CF3)4 cyclobutadienyl ligand with approximate C4v local symmetry and a eta5-C5H5 cyclopentadienyl ring. The centroids of the rings and the S atoms of a chelating dithiocarbamate ligand define the pseudo-tetrahedral coordination of the Mo atom. The Mo-C(cyclobutadienyl) bond lengths [2.189 (2)-2.211 (2) A] are unusually short, probably reflecting strong electron withdrawal by the trifluoromethyl groups. The molecules straddle crystallographic mirror planes. PMID- 15805630 TI - 2,5-dichloro-2,5-dihydrothiophene 1-oxide: determination of the complete stereochemistry. AB - The title five-membered heterocycle, C4H4Cl2OS, adopts an envelope conformation with the S atom at the tip of the flap. All three ring substituents, viz. the sulfoxide O atom and the two Cl atoms, are cis to each other. The two C atoms alpha to the sulfoxide group are also bonded to chlorine. The electron withdrawing chlorine substituents give rise to weak C-H...O hydrogen bonds with the sulfoxide O atom of a symmetry-related molecule [H...O = 2.44 (2) and 2.61 (2) A, C...O = 3.143 (3) and 3.302 (2) A and C-H...O = 129.9 (19) and 135.1 (19) degrees ]. There is also a possible weak C-H...Cl interaction. Chains of molecules held together by these weak interactions run parallel to the a axis. PMID- 15805631 TI - Di-mu-halogeno-bis[halogeno(triphenylphosphine)mercury(II)], [Ph3PHgX(mu X)2XHgPPh3], reinvestigated at 120 K for X = Cl and Br, and a second polymorph for X = I, also at 120 K. AB - Di-mu-chloro-bis[chloro(triphenylphosphine)mercury(II)], [Hg2Cl4(C18H15P)2], (I), and di-mu-bromo-bis[bromo(triphenylphosphine)mercury(II)], [Hg2Br4(C18H15P)2], (II), have been reinvestigated at 120 K. The molecules of (I) lie across inversion centres in space group P2(1)/n, and in both (I) and (II) the complexes are linked into three-dimensional frameworks by a combination of C-H...X (X = Cl and Br) and C-H...pi(arene) hydrogen bonds. At 120 K, di-mu-iodo bis[iodo(triphenylphosphine)mercury(II)], [Hg2I4(C18H15P)2], (III), crystallizes as a new polymorphic form having Z' = 1/2, where the complexes lie across inversion centres in space group P2(1)/n; the complexes are linked into sheets by a combination of C-H...I and C-H...pi(arene) hydrogen bonds. In the Z' = 1 polymorph of this compound, a single C-H...I hydrogen bond generates simple chains. PMID- 15805632 TI - 2-(3-benzoyl-1-pyridinio)-3,4-dioxocyclobutenolate. AB - The title compound, C16H9NO4, also known as the 3-benzoylpyridinium betaine of squaric acid, exhibits a dipolar electronic ground-state structure with a positively charged pyridinium fragment and a negatively charged squarate moiety. In the molecule, the two aromatic rings are twisted by 56.03 (2) degrees relative to one another. The three-dimensional packing of the molecules is stabilized by C H...O short contacts. PMID- 15805633 TI - Three isomeric N-(nitrophenyl)succinimides: isolated molecules, hydrogen-bonded sheets and a hydrogen-bonded three-dimensional framework. AB - Molecules of N-(2-nitrophenyl)succinimide, C10H8N2O4, are linked into sheets by a combination of C-H...O and C-H...pi(arene) hydrogen bonds. Molecules of N-(3 nitrophenyl)succinimide are linked into a three-dimensional framework by a combination of a two-centre C-H...O hydrogen bond and a three-centre C-H...(O)2 hydrogen bond. Molecules of N-(4-nitrophenyl)succinimide which lie across twofold rotation axes in space group C2/c, participate in no direction-specific intermolecular interactions. PMID- 15805634 TI - Trifluoromethyl derivatives of pentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecane. AB - Three cage-like polycyclic compounds, viz. exo-8 (trifluoromethyl)pentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecan-endo-8-ol, C12H13F3O, 5-(trifluoromethyl)-4 oxahexacyclo[5.4.1.0(2,6).0(3,10).0(5,9).0(8,11)]dodecan-3-ol, C12H11F3O2, and N [exo-11-(trifluoromethyl)-endo-11 (trimethylsilyloxy)pentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecan-8 ylidene]aniline methanol solvate, C21H24F3NOSi.CH4O, were obtained from the corresponding oxo derivatives by nucleophilic trifluoromethylation with (trifluoromethyl)trimethylsilane in 1,2-dimethoxyethane solution in the presence of CsF. The crystal structures show that the addition of trifluoromethanide occurs exclusively from the exo face of the polycyclic ketones. Further examination of the crystal structures, together with that of the starting pentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecane-8,11-dione, C11H10O2, showed that increasing substitution at the 8- and/or 11-positions in the cage molecules increases the non-bonded intramolecular C...C distances at the mouth of the cage and changes the puckering of the five-membered rings involving the 8- and 11 positions from an envelope towards a distorted half-chair conformation. Intermolecular co-operative O-H...O hydrogen bonds in the endo-8-ol compound link the molecules into tetramers. PMID- 15805635 TI - 9-(2,6-dichlorophenoxycarbonyl)-10-methylacridinium trifluoromethanesulfonate and its precursor 2,6-dichlorophenyl acridine-9-carboxylate. AB - The title compounds, C21H14Cl2NO2+.CF3O3S-, (I), and C20H11Cl2NO2, (II), form triclinic crystals. Adjacent cations of (I) are oriented either parallel or antiparallel; in the latter case, they are related by a centre of symmetry. Together with the CF3SO3- anions, the antiparallel-oriented cations of (I) form layers in which the molecules are linked via a network of C-H...O and pi-pi interactions (between the benzene rings). These layers, in turn, are linked via a network of multidirectional pi-pi interactions between the acridine rings, and the whole lattice is stabilized by electrostatic interactions between ions. Adjacent molecules of (II) are oriented either parallel or antiparallel; in the latter case, they are related by a centre of symmetry. Parallel-oriented molecules are arranged in chains stabilized via C-H...Cl interactions. These chains are oriented either parallel or antiparallel and are stabilized, in the latter case, via multidirectional pi-pi interactions and more generally via dispersive interactions. Acridine and independent benzene moieties lie parallel in the lattices of (I) and (II), and are mutually oriented at an angle of 33.4 (2) degrees in (I) and 9.3 (2) degrees in (II). PMID- 15805637 TI - N-(triphenylmethylsulfanyl)phthalimide. AB - The title compound, C27H19NO2S, contains chains of fused R(2)(2) (19) rings formed by intermolecular C-H...O=C hydrogen bonds and running along the [011] and [0-11] directions. These chains are linked through short intermolecular C-H...pi contacts, giving rise to sheets. The conformation of the title compound is dominated primarily by the nearly orthogonal lone-pair orbitals on the N and S atoms. PMID- 15805636 TI - 2-(Acridin-9-ylimino)-3-dimethylamino-1,3-thiazolidin-4-one. AB - In the title compound, C18H16N4OS, prepared by the reaction of 4-(acridin-9-yl) 1,1-dimethylthiosemicarbazide with methyl bromoacetate, the acridine and thiazolidine ring systems are both non-planar and, because of steric requirements, almost perpendicular, with a dihedral angle between their planes of 99.69 (6) degrees. C-H...O and C-H...pi(arene) hydrogen bonds stabilize the crystal structure in the solid state. PMID- 15805638 TI - (2Z)-3-(5-Hydroxy-4-oxo-4H-chromen-3-yl)acrylonitrile. AB - The title compound, C12H7NO3, consists of a chromone moiety substituted in position 3 with an acrylonitrile group in a Z configuration. The two planar groups are twisted with respect to one another. The only significant hydrogen bond in the structure is an intramolecular O-H...O bond. pi-pi contacts connecting aromatic groups and C-H...O intermolecular weak interactions lead to a supramolecular layer arrangement. PMID- 15805639 TI - 2,6-dimethyl-1,4-benzoquinone 4-monooxime. AB - Molecules of the title compound, C8H9NO2, are linked into sheets by a combination of C-H...N, O-H...N and O-H...O hydrogen bonds and C-H...pi interactions. The hydrogen bonds are arranged as described by the graph-set ring notations R(2)(2)(7) and R(3)(3)(5), and a C8 chain motif. There are two planar symmetry independent molecules in the asymmetric unit, with a dihedral angle of 19.24 (5) degrees between their least-squares mean planes. PMID- 15805640 TI - N-cinnamoylsaccharin. AB - The title compound [systematic name: 2-cinnamoyl-1,2-benzisothiazol-3(2H)-one 1,1 dioxide], C16H11NO4S, contains both saccharin and cinnamoyl groups. The molecule is approximately planar in the solid state, and adjacent molecules are connected by C-H...O and C-H...pi(phenyl) interactions. In the C-H...pi interaction, the C...CgA distance is 3.916 (4) A (CgA is the non-fused benzene ring centroid) and the C-H...pi angle is 156 (2) degrees . A feature of the molecular geometry is the narrow C-S-N angle of 92.51 (9) degrees in the five-membered ring. This angle relieves strain from the ring and makes it possible for the whole saccharin group to become quite planar. PMID- 15805641 TI - 3-iodoanilinium 2-carboxy-6-nitrobenzoate: a three-dimensional framework built from O-H...O and N-H...O hydrogen bonds and a two-centre iodo-nitro interaction. AB - In the title compound, C6H7IN+.C8H4NO6-, the anions are linked by a single type of O-H...O hydrogen bond into C(7) chains, and these chains are linked via three independent N-H...O hydrogen bonds into sheets. The sheets, in turn, are linked by a two-centre iodo-nitro interaction into a single three-dimensional framework. PMID- 15805642 TI - Two (+)-alpha,4-dimethyl-2-oxocyclohexaneacetic acids: hydrogen bonding in a terpenoid gamma-keto acid and in a diastereomeric lactol. AB - The (+)-(alphaS,1S,4R)-diastereomer of the title structure, C10H16O3, aggregates in the solid as non-symmetric dimers with disorder in both carboxyl groups [O...O = 2.710 (5) and 2.638 (5) A]. The two molecules constituting the asymmetric unit pair around a pseudo-twofold rotational axis and differ only slightly in their distances and angles, but one methyl group displays rotational disorder absent in the other molecule. Five intermolecular C-H...O close contacts exist, involving both ketone groups. The (+)-(alphaR,1R,4R)-diastereomer exists in the crystal in its closed-ring lactol form, (3R,3aR,6R,7aR)-2,3,3a,4,5,6,7,7a-octahydro-7a hydroxy-3,6-dimethylbenzo[b]furan-2-one, C(10)H(16)O(3), and aggregates as hydrogen-bonded catemers that extend from the hydroxyl group of one molecule to the carbonyl group of a neighbor screw-related along b [O...O = 2.830 (3) A and O H...O = 169 degrees]. One close intermolecular C-H...O contact exists involving the carbonyl group. PMID- 15805643 TI - A diaryl-terminated hexa-1,5-diyne-3,4-dione. AB - The structure of bis(4-tert-butyl-2,6-dimethylphenyl)hexa-1,5-diyne-3,4-dione, C30H34O2, has been determined, revealing an extended s-trans conformation of the dione and the two ynone moieties, which are shielded by the flanking methyl substituents. The structural parameters and the packing arrangement suggest little electronic delocalization between the two ynone moieties. PMID- 15805644 TI - threo-2-(2,6-dimethoxyphenoxy)-1-(4-hydroxy-3,5-dimethoxyphenyl)propane-1,3-diol: a conformational study. AB - In the crystal structure of the title compound, C19H24O8, the molecules adopt a conformation in which the bulky 2,6-dimethoxyphenoxy and 4-hydroxy-3,5 dimethoxyphenyl groups are almost as far apart as possible. The C(aryl)...C(aryl) distance is 4.8766 (19) A, which is close to the calculated maximum value (4.92 A). The C(aryl)-C-C-O(aryloxy) torsion angle is 173.76 (11) degrees and the C(benzylic)-C-O-C(aryl) torsion angle is 149.09 (11) degrees. The conformation is compared with those of related lignin model compounds. The hydrogen-bonding pattern is discussed in terms of graph-set theory. PMID- 15805645 TI - Packing effects in 4,4'-bis(4-hydroxybutyl)-2,2'-bipyridine and 4,4'-bis(4 bromobutyl)-2,2'-bipyridine. AB - Structure analyses of 4,4'-bis(4-hydroxybutyl)-2,2'-bipyridine, C18H24N2O2, (I), and 4,4'-bis(4-bromobutyl)-2,2'-bipyridine, C18H22Br2N2, (II), reveal intermolecular hydrogen bonding in both compounds. For (I), O-H...N intermolecular hydrogen bonding leads to the formation of an infinite two dimensional polymer, and pi stacking interactions are also observed. For (II), C H...N intermolecular hydrogen bonding leads to the formation of a zigzag polymer. The two compounds crystallize in different crystal systems, but both molecules possess Ci symmetry, with one half molecule in the asymmetric unit. PMID- 15805646 TI - N-[(R)-1-(2-hydroxy-5-methylphenyl)ethyl]-N-[(R)-1-(2-methoxy-5-methylphenyl)-2 phenylethyl]aminium chloride. AB - The title compound, C25H30NO2+.Cl-, has been synthesized, and the crystal structure shows that it is mainly stabilized through intermolecular N-H...Cl and O-H...Cl and intramolecular N-H...O hydrogen bonds. The absolute configuration of the new stereogenic center (the C atom adjacent to the N atom on the phenol side) was determined to have an R configuration. PMID- 15805647 TI - Ethyl 5-isopropoxy-4-methyl-beta-carboline-3-carboxylate: structural determinants of benzodiazepine-receptor antagonism. AB - Molecules of the title compound, C18H20N2O3, are linked into ribbons by N-H...O and N-H...N hydrogen bonds. Stereochemical comparison with Ro 15-1788 (viz. ethyl 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine-3 carboxylate) has identified three electronegative N and O atoms in the molecule as features likely to be responsible for its activity as a benzodiazepine receptor antagonist. PMID- 15805648 TI - Arteriovenous malformations: current status of surgery. PMID- 15805649 TI - Management of arteriovenous malformations: a surgical perspective. AB - The management strategies for arteriovenous malformations (AVMs) continue to evolve, spurred by advancing technology and improved understanding of the natural history of these lesions. In general, intervention is reserved for Spetzler Martin Grade I-III lesions or for those with certain high-risk features. Grade IV V AVMs, in contrast, are usually managed conservatively. Although multimodality therapy incorporating endovascular and/or radiosurgical techniques is increasingly common, microsurgical removal remains the definitive form of treatment. PMID- 15805650 TI - Genetic therapy in gliomas: historical analysis and future perspectives. AB - High-grade gliomas are relatively frequent in adults, and consist of the most malignant kind of primary brain tumor. Being resistant to standard treatment modalities such as surgery, radiation, and chemotherapy, it is fatal within 1 to 2 years of onset of symptoms. Although several gene therapy systems proved to be efficient in controlling or eradicating these tumors in animal models, the clinical studies performed so far were not equally successful. Most clinical studies showed that methodologies that increase tumor infection/transduction and, consequently confer more permanent activity against the tumor, will lead to enhanced therapeutic results. Due to the promising practical clinical benefits that can be expected for the near future, an exposition to the practicing neurosurgeon about the basic issues in genetic therapy of gliomas seems convenient. Among the main topics, we shall discuss anti-tumoral mechanisms of various genes that can be transfected, the advantages and drawbacks of the different vectors utilized, the possibilities of tumor targeting by modifications in the native tropism of virus vectors, as well as the different physical methods for vector delivery to the tumors. Along with the exposition we will also review of the history of the genetic therapy for gliomas, with special focus on the main problems found during the advancement of scientific discoveries in this area. A general analysis is also made of the present state of this promising therapeutic modality, with reference to the problems that still must be solved and the new paradigms for future research in this area. PMID- 15805651 TI - Telemedicine in neurology: underutilized potential. AB - Advances in telecommunication which started with telephone lines, FAX, integrated service digital network (ISDN) lines and now internet have provided an unprecedented opportunity for transfer of knowledge and sharing of information. The information can be used for overlapping applications in patient care, teaching and research. In medicine there is increasing utilization of telemedicine; radiology and pathology being regarded as mature specialties and emergency medicine as maturing specialties compared to other evolving specialties which include psychiatry, dermatology, cardiology and ophthalmology. Of the emergencies, status epilepticus and stroke have high potential for improving patient management. Administration of tPA was more frequent when carried out under telemedicine guidance. Telemedicine has great potential for medical education. The principles of education are in congruence with those of telemedicine and can be closely integrated in the existing medical education system. Our experience of telemedicine as a medical education tool is based on video conferencing with SCB Medical College, Cuttack. We had 30 sessions during 2001 to 2004 in which 2-3 cases were discussed in each session. The patients' details, radiological and neurophysiological findings could be successfully transmitted. These conferences improved the knowledge of participants, provided an opportunity for a second opinion as well as modified the treatment decisions in some cases. The advances in telemedicine should be utilized more extensively in neurology, especially in emergency management, epilepsy and stroke patients as well, as it may have a role in neurophysiology and movement disorders. PMID- 15805652 TI - Role of clinical neurophysiological tests in evaluation of erectile dysfunction in people with spinal cord disorders. AB - BACKGROUND: While erectile dysfunction is frequent among people with disorders of the spinal cord, the role of various clinical neurophysiological tests in assessment is not clear. AIMS: To study the role of clinical neurophysiological investigations in assessing erectile dysfunction among men with spinal cord disorders. SETTING: National Institute of Mental Health and Neurosciences, India. DESIGN: Survey. MATERIALS AND METHODS: Subjects with a score of 21 or less on the International Index of Erectile Function-5 were classified as with erectile dysfunction and with a score of more than 21 as without erectile dysfunction. Clinical neurophysiological studies done were Sympathetic Skin Response from limbs, posterior tibial sensory evoked potential, pudendal sensory potential and bulbocavernous reflex. STATISTICAL ANALYSES: Chi-square test. RESULTS: Among 40 subjects 26 had erectile dysfunction. The frequency of abnormalities in clinical neurophysiological studies were: pudendal sensory evoked potentials--16, posterior tibial sensory evoked potentials--26, bulbocavernous reflex--5, sympathetic skin response from sole--24 and, sympathetic skin response from palm- 18. Significant associations were noted between erectile dysfunction and abnormal pudendal sensory evoked potentials (P=0.0479), and absent sympathetic skin response from palm (P=0.0279) and sole (P< 0.001). There was no correlation between erectile dysfunction and posterior tibial sensory evoked potentials (P=0.133) or bulbocavernous reflex (P=0.418). Sympathetic skin response from sole was most sensitive (80.8%) and had best positive (87.5%) and negative predictive (68.8%) values. The specificity of these three tests was 78.6%. CONCLUSIONS: Sympathetic skin response from the sole of the foot was the most sensitive and specific clinical neurophysiological test for erectile dysfunction in spinal cord disorders. PMID- 15805653 TI - Myasthenic crisis: clinical features, complications and mortality. AB - BACKGROUND AND OBJECTIVE: Myasthenic crisis is a life-threatening complication of myasthenia gravis (MG) and when treated aggressively is associated with good outcome. MATERIALS AND METHODS: Retrospective study of case records of patients with episodes of myasthenic crisis. RESULTS: Twenty-one (22%) of the 95 patients with MG (9 with thymoma), experienced 23 episodes of myasthenic crisis, 3 (33%) in patients with thymoma. The crisis episodes occurred within 2 years of disease onset in 11 (52%) patients. Infection was the most common primary precipitant of the crisis occurring in 65%. The median duration of the crisis episode was 11 days (7-39 days), and the median neurological intensive care unit stay was 15 days (range 9-47 days). Fifteen (65%) episodes were treated with small volume plasma exchange (PE) and 8 (35%) episodes received intravenous immunoglobulin (IVIg). The time taken for disease stabilization, the median number of days for extubation, was 8 days (range 7-12) in the PE group and 10 days (range 7-39) in the IVIg group. Disease stabilization could not be achieved in one patient in the IVIg group. Ventilator-associated pneumonia (VAP) was the commonest complication, seen in 30%. Two (8%) of the 23 episodes of crisis were fatal, one resulting from VAP and septicemia, and the other due to crisis itself. All the 19 patients who survived to discharge had complete resolution of admission symptoms. CONCLUSIONS: In patients with myasthenic crisis, both therapeutic options, PE and IVIg, are equally effective in disease stabilization. To achieve good outcomes all efforts should be directed at decreasing the duration of intubation and also aggressively treating the associated medical complications. PMID- 15805654 TI - Surgery of vestibular schwannomas: an institutional experience. AB - AIMS: To report management results of vestibular schwannomas (VS) treated surgically in our institute, with particular reference to completeness of tumor excision, facial nerve and hearing preservation and complications of surgery. STUDY DESIGN AND SETTINGS: Retrospective study of 259 patients treated during the years 1988 to 2002. MATERIALS AND METHODS: The facial nerve function and hearing assessment was done according to House-Brackmann [HB] grading and pure tone audiometry (PTA) respectively. All patients were operated by retro-mastoid sub occipital approach. RESULTS: Most patients had large tumors and had no useful hearing (90%), had disabling cerebellar ataxia (88%) and presented with features of raised intra-cranial pressure (45%). Large sized tumors were in 41.3% and giant sized tumors were in 56% cases. Complete tumor excision was carried out in 96.5% and anatomical preservation of facial nerve was achieved in 79.2% cases. Hearing preservation was achieved in 8 patients. Cerebrospinal fluid leak with or without meningitis and transient lower cranial nerve paresis were common complications. The mortality was 6%. CONCLUSIONS: With experience, complete tumor excision with good facial nerve preservation can be achieved in large tumors. Hearing preservation is difficult in bigger tumors. Prevention and control of infection was a major concern. PMID- 15805655 TI - Frequent seizures and polytherapy can impair quality of life in persons with epilepsy. AB - CONTEXT: Careful evaluation of pharmacotherapy, seizure control and quality of life (QOL) are helpful in improving epilepsy care but such data are relatively meager from developing countries. AIMS: To audit pharmacotherapy, seizure control and QOL in persons with epilepsy and to identify factors associated with impaired QOL. SETTINGS AND DESIGN & MATERIALS AND METHODS: The study was carried out using a cross-sectional design in the setting of a tertiary care epilepsy center in India. Persons with epilepsy with > 12 months follow-up at this Center and aged > 16 years were eligible for enrollment. Persons with other disabilities or pregnancy were excluded. Subjects were interviewed with a standard questionnaire and an adapted version of Quality of Life in Epilepsy - 31 (QOLIE-31). Data pertaining to treatment at the time of referral to this center was extracted from medical records. STATISTICAL ANALYSIS USED: Chi-square test, analysis of variance and multiple regression analysis were carried out for statistical significance. RESULTS: One hundred and twelve patients with epilepsy (59 males, mean age 31.2+/ 10.7 years) were included. Forty-seven (42%) persons had Generalized Epilepsy (GE) and 65 persons (58%) had Localization-Related Epilepsy (LRE). At entry 24 persons (21.4%) were not on treatment and 59 persons (64.8%) had frequent seizures. At last follow-up 64 persons (57.1%) were seizure-free, 83 persons (74.1%) were on monotherapy and 29 were (25.9%) on polytherapy. Cost of drug at entry was INR 2276 (monotherapy) and INR 3629 (polytherapy) (45 INR = 1 USD). At the time of last follow-up, it was 1898 and 4929 respectively. QOLIE-31 Total Score (TQOL) ranged from 22.6 to 94.4 (mean 68.0 +/- 15.8). Multiple regression analysis showed significant correlation between low TQOL score and polytherapy (P=0.002) and occurrence of one or more seizures per month (P=0.001). CONCLUSIONS: Frequent seizures and polytherapy are associated with lower QOL in persons with epilepsy. PMID- 15805656 TI - The effects of hematocrit and age on transcranial Doppler measurements in patients with recent ischemic stroke. AB - BACKGROUND: The effects of age and hematocrit on transcranial Doppler (TCD) velocities have not been evaluated in a large patient group with recent ischemic stroke. AIM: This study assessed the effects of age and hematocrit on TCD measurements in patients with recent ischemic stroke compared to patients with non-vascular diseases. SETTINGS AND DESIGN: University Hospital, retrospective study. MATERIALS AND METHODS: TCD records and data files of 862 consecutive patients (mean age, 57+/-16 years) with various neurological diagnoses were reviewed retrospectively. The peak systolic, end diastolic and mean flow velocities (FV), systolic/diastolic ratios and pulsatility indices (PI) in the middle cerebral arteries were averaged and the effect of age and hematocrit values on these TCD values was studied. Independent samples t test, Pearson's coefficients of correlation, and linear regression test were used for statistical analysis. RESULTS: Among 862 patients, 413 were women (mean age, 53+/-17 years) and 449 were men (mean age, 60+/-13 years). Peak systolic and mean FV were higher and hematocrit concentration was lower in women compared to men (P< 0.001). The relation of TCD velocities with age and hematocrit was more remarkable in the group of patients with non-vascular neurological disorders. PI values demonstrated a significant correlation to age (r=+0.47) (P< 0.001), but did not change significantly with hematocrit level. CONCLUSIONS: It should be remembered that blood FV measured by TCD may be significantly affected by age and hematocrit level. However, there seems to be no significant association between TCD velocities and hematocrit in patients with a recent ischemic stroke. PMID- 15805657 TI - Clinical and laboratory features and response to treatment in patients presenting with vitamin B12 deficiency-related neurological syndromes. AB - AIMS AND OBJECTIVES: To study the clinical and laboratory features of patients admitted with vitamin B12 deficiency-related (B12def) neurological syndromes. SETTINGS AND DESIGN: A hospital-based retrospective and prospective study conducted at a referral teaching hospital. MATERIALS AND METHODS: Consecutive patients admitted with vitamin B12 deficiency-related neurological disorders during a three-year period from June 2000 to May 2003 were included. Data regarding clinical and laboratory features were obtained. Follow-up was done at least six months following treatment with parenteral vitamin B12. Chi-square test was used for statistical analysis. RESULTS: A total of 63 patients (52 males) with a mean age of 46.2 years were studied. The mean duration of symptoms at presentation was 10.3 months. Myeloneuropathy (54%) was the commonest neurological manifestation, followed by myeloneuropathy with cognitive dysfunction (34%), and peripheral neuropathy (9%). Neuropsychiatric manifestations and dementia were observed in 38% and 19% of patients respectively. All the patients had megaloblastic changes in the bone marrow smear. Eleven (17.5%) patients had both hemoglobin and the mean corpuscular volume (MCV) within the normal range. Follow-up after at least six months of therapy with parenteral B12 showed improvement in 54% patients. CONCLUSIONS: A high index of suspicion of B12def is required in patients presenting with myelopathy, cognitive decline, or neuropathy. A normal hemoglobin or MCV does not exclude B12def; therefore, other tests such as bone marrow smear and serum vitamin B12 assay are essential, as the condition is often reversible with treatment. PMID- 15805658 TI - The PHC-cog: a brief cognitive function screening test for the elderly. AB - BACKGROUND: Cognitive dysfunctions such as dementia have emerged as a leading public health problem among the elderly. Its early detection is important for the management of patients, and for the educational support of other family members. Although cognitive dysfunction screening tests are available, they have not been widely utilized in community or primary care settings. AIM: To validate the PHC cog (Public Health Center Cognitive Dysfunction Test) and to assess it as a simple and brief method for massive screening of cognitive dysfunctions in the primary care setting. MATERIALS AND METHODS: The study comprised 137 community dwelling patients with dementia and 134 age, sex, and education-matched controls. The PHC-cog was made combining patients' and informants' questionnaires to improve performance. The PHC-cog patient's section briefly assessed the patient's cognition. RESULTS: The PHC-cog Patient's Section had a sensitivity and specificity of 0.75 and 0.92, respectively. The PHC-cog Informants' Section had a sensitivity and specificity of 0.79 and 0.83, respectively. The total method of administering the PHC-cog had a sensitivity and specificity of 0.96 and 0.82, and the two-stage method had a sensitivity and specificity of 0.92 and 0.76, respectively. CONCLUSIONS: The PHC-cog test is a simple, accurate and reliable, performance-based tool in the screening for cognitive dysfunction. The PHC-cog test is quick, and easy-to-use, and will hopefully become widely used in the cognitive screening of the aging population in the primary public medical institutions of Korea. PMID- 15805659 TI - Electro-clinical characteristics and postoperative outcome of medically refractory tumoral temporal lobe epilepsy. AB - BACKGROUND: Very few studies have specifically addressed surgical treatment and outcome of patients with tumor-related temporal lobe epilepsy (TLE). AIM: To define the postoperative seizure outcome and the factors that influenced the outcome of patients with tumor-related TLE. MATERIALS AND METHODS: We selected patients whose surgical pathology revealed a temporal lobe neoplasm and who had completed > 1 year of postoperative follow-up. We reviewed the clinical, EEG, radiological and pathological data, and the seizure outcome of these patients and assessed the factors that influenced the outcome. RESULTS: Out of the 409 patients who underwent surgery for refractory TLE during the 8-year study period, there were 34 (8.3%) patients with temporal lobe neoplasms. The median age at surgery was 20 years and the median duration of epilepsy prior to surgery was 9.0 years. MRI revealed tumor in the mesial location in 21 (61.8%) patients. Interictal and ictal epileptiform EEG abnormalities were localized to the side of th lesion in the majority. Mesial temporal lobe structures were included in the resection, if they were involved by the tumor; otherwise, lesionectomy alone was performed. During a median follow-up of 4 years, 27 (79%) patients were completely seizure-free. The only factor that predicted long-term seizure-free outcome was being seizure-free during the first two postoperative years. CONCLUSIONS: Our results emphasize the fact that in patients with tumoral TLE, when the seizures are medically refractory, surgery offers potential for cure of epilepsy in the majority. PMID- 15805660 TI - Cerebral computed tomographic angiography scan delay in subarachnoid hemorrhage. AB - CONTEXT: Computed tomographic angiography (CTA) is widely applied in the evaluation of cerebral vessels. Contrast enhancement in cerebral CTA without care or test bolus is not always sufficient for high-quality images. AIMS: Evaluation of the possibilities of calculation of scan delay for cerebral CTA in case of subarachnoid hemorrhage (SAH), based on clinical data of a patient and to find out prognostic error of the model. SETTINGS AND DESIGN: Prospective study in Neurosurgery and Radiology departments. MATERIALS AND METHODS: Scan delay in 53 patients suffering an acute SAH was measured employing test bolus technique. Cerebral CTA was performed afterwards. STATISTICAL ANALYSIS USED: SPSS for Windows v.10.1 software package was applied for dispersion analysis, including one-sample Kolmogorov-Smirnov's test and Levene's Test of Equality of Error Variances. RESULTS: A statistical model for the prediction of scan delay in SAH was developed. Cerebral CTA scan delay was dependent upon age, neurological status and impact of the latter factors together (P< 0.05). The determined mean square error of prognosis of scan delay of the developed model equals 3.3 sec. CONCLUSION: Using our proposed model it is possible to estimate an optimal delay time for CTA in most patients with SAH with a determined error. PMID- 15805661 TI - Effect of naloxone on aluminum-induced learning and memory impairment in rats. AB - BACKGROUND: Uptake of aluminum may disturb the learning and memory of humans or animals. Naloxone (NAL) has been shown to exert beneficial effects on memory deficits. AIMS: We investigated the effects of naloxone on aluminum-induced learning and memory impairment in rats. SETTINGS AND DESIGN: Aluminum-induced learning and memory impairment model was established by gavage of Aluminum chloride (600 mg/kg) for 3 months. Rats were divided into three groups viz. naloxone-treated rats (NAL 0.8 mg/kg, i.p. daily for 7 days), non-treated model rats and normal controls. MATERIALS AND METHODS: The Morris water maze test was performed to study spatial learning and memory. Long-term potentiation (LTP) of the Schaffer collateral-CA1 synapse was recorded. Aluminum and zinc contents in the hippocampus were assayed with atomic absorption spectrophotometry. STATISTICAL ANALYSIS: Parameters of the hidden and visible platform trials and data of LTP were analyzed using two-way repeated measures ANOVA. RESULTS: In the hidden platform trials, escape latencies of the NAL rats were significantly shorter than that of the non-treated rats (P=0.000, 95% confidential interval low bound 14.31, upper bound 22.68). In probe trails, the number of entries in the target area of the NAL rats (6.75+/-1.28 times/min) was more than that of non treated model rats (4.56+/-2.16 times/min, P=0.004, 95% confidence interval low bound -3.65, upper bound -0.788). The magnitudes of LTP recorded in the CA1 pyramidal neurons of the NAL-treated rats were significantly augmented when compared to the non-treated model rats (P=0.005, 95% confidence interval low bound 0.16, upper bound 0.84). CONCLUSIONS: NAL could facilitate spatial learning and memory and enhance LTP in the CA1 region of the hippocampus in aluminum induced learning and memory impairment in rats. PMID- 15805662 TI - Anterior instrumentation for cervical spine tuberculosis: an analysis of surgical experience with 61 cases. AB - OBJECTIVE: To evaluate the efficacy of anterior instrumentation in patients with subaxial and cervicodorsal spinal tuberculosis in reconstruction of the spine, providing pain relief, neurological recovery and prevention of deformity. MATERIALS AND METHODS: The records of 61 consecutive patients, of surgically treated spinal tuberculosis affecting C3 to D2 region, in our neuro and spinal surgery unit over a five-year period were retrospectively reviewed. Patients with involvement of the C3-C6 vertebrae underwent excision of the involved vertebrae and intervertebral discs followed by reconstruction with titanium implants by anterior approach. A transclavicular approach was used for patients with involvement of the C7-D2 vertebrae. A five-drug antituberculous regimen was administered for a period of one year. The follow-up ranged from 24 to 84 months (mean 38 months). Clinical and radiological assessment using flexion and extension radiographs was performed at 24 months for all cases. RESULTS: The neck pain score based on a visual analog scale (1-10) changed from a pre-operative average of 7 to 2 at follow-up after 4 months. Fifty-two patients (85%) had complete relief of pain while 16 patients who had Grade III to IV muscle strength regained complete power. The asymmetric wasting in patients with involvement of the cervicodorsal region did not recover completely. Flexion-extension radiographs at 24 months did not show any evidence of instability or nonunion. CONCLUSIONS: Anterior reconstruction using titanium plates and locking screws for stabilization of the subaxial and cervicodorsal region tuberculosis is a useful adjunct in preventing kyphotic deformity. A satisfactory segmental stability and fusion is achieved by this technique. PMID- 15805663 TI - Protective effects of cisternal irrigation on leptomeningeal and cortical structures in meningitis: an experimental study. AB - BACKGROUND: Meningitis, termed as foreign material collection in the subarachnoid space, leads to various meningeal, cerebral and spinal cord pathologies. Meningitis still remains a problematic disease with severe complications in spite of advanced medical technology. AIMS: In this study, we aimed to investigate the role of cisternal irrigation in the prevention of meningitis complications. SETTING AND STUDY DESIGN: Experimental study was done in the Social Security Hospital of Erzurum. Histopathological specimens were evaluated in the Pathology Department in the Ataturk University Research Hospital, Erzurum, Turkey. MATERIALS AND METHODS: This study was conducted on twelve lambs. Experimental meningitis was achieved with streptococcus pneumonia. Two animals were not treated. Ten animals were given Cefotaxime (4 x 1g/day) for 20 days, and additionally half of these animals underwent cisternal irrigation. Then, all animals were sacrificed and brains were observed histopathologically. RESULTS: Massive purulent CSF formation, hemorrhagic cortical lesions, vascular congestion, leptomeningeal and cortical adhesions and brain edema were observed in the non-irrigated group, but these findings were observed slightly or absent in the irrigated group. CONCLUSION: Meningitis can affect all central neural tissues, consequently serious central nervous system lesions may develop. The irrigation procedure may decrease the percentage and severity of meningitis complications by way of the excretion of inflamed purulent collection from the subarachnoid spaces. PMID- 15805664 TI - Genetic epidemiology of epilepsy: a twin study. AB - The study explored the genetic susceptibility and prevalence of epilepsy in twins. The data on epilepsy were retrieved from the health records of 199 pairs of twins. Proband concordance rate in monozygotic (MZ) twins was four times more than that in dizygotic (DZ) twins (0.67 vs. 0.17). Three of 15 (20%) affected twin kinships had epileptic first-degree relatives. These findings indicated significant underlying genetic susceptibility to epilepsy with the Holzinger's heritability estimate being 0.45. The prevalence of epilepsy was similar in MZ (45.45), DZ (45.11) twins, and their non-twin siblings (47.60). In the general population from various nationalities, the mean prevalence rate of epilepsy varied from 5 to 17 per 1000. The appreciably higher prevalence rate in twin kinships could be attributed to peculiar development factors associated with the twinning process or the intrauterine environment of mothers having tendencies to bear twins. Of the genetic markers, PTC locus seemed to be associated with the susceptibility to epilepsy. The allele frequency of non-tasters (t) seemed greater in epileptic twin kinships (0.71) than that in the general population (0.53). The frequency of non-tasters was similar in MZ and DZ twins and singletons: 27.3%, 26%, and 27.7% respectively. The PTC data on the general population was based on a sample of 278 individuals. PMID- 15805665 TI - Decreased electroencephalogram alpha band [8-13 Hz] power in amyotrophic lateral sclerosis patients: a study of alpha activity in an awake relaxed state. AB - An attempt was made to quantitatively analyze the alpha activity in the awake relaxed state of Amyotrophic Lateral Sclerosis (ALS) patients and was compared with normals. ALS patients showed significantly low amplitude with a corresponding alpha band (8-13 Hz) power reduction, in both hemispheres though the change was more prominent in the left hemisphere. A review of the literature revealed no studies done on alpha oscillations in ALS patients; hence the results may have important implications for the interpretation of resting state brain activities. PMID- 15805666 TI - Hallervorden spatz disease: MR and pathological findings of a rare case. AB - We describe a child with pathologically proven Hallervorden Spatz disease. He presented with extrapyramidal symptoms and characteristic "eye-of-the-tiger" sign on magnetic resonance imaging. He was given the possible benefit if any of deep brain stimulation with no much improvement. Pathological examination of the brain showed iron deposition in bilateral globus pallidi, spongiform change and neuron axonal degeneration (spheroids). PMID- 15805667 TI - Giant pituitary adenoma invading the clivus. AB - A 26-year-old man was admitted with complaints of worsening of vision for one month. Investigations revealed a large sellar and parasellar tumor mass invading into both the cavernous sinuses. The tumor invaded the clivus and both petrous apices. It was radically but partially resected through a transsphenoidal approach. Following surgery, the patient showed symptomatic improvement. The residual tumor was treated by radiation treatment. The case is discussed and the literature on the subject is briefly reviewed. PMID- 15805668 TI - Acute myopia induced by topiramate: report of a case and review of the literature. AB - Topiramate, a new anticonvulsant, is also used for the prophylaxis of migraine and cluster headache. A serious but not often discussed side effect of the drug is the development of acute myopia and acute angle-closure glaucoma in the early stage of therapy that subsides rapidly with prompt discontinuation. One such case is reported here and the relevant literature in this regard is also reviewed. PMID- 15805669 TI - Rosai-Dorfman disease mimicking a sphenoid wing meningioma. AB - A 40-year-old male presented with a single generalized tonic-clonic seizure. MRI revealed an enhancing, dural-based, left lateral sphenoid wing lesion suggestive of a meningioma. At microsurgical excision, the lesion was firm and relatively avascular. The histopathological report revealed S-100 positive histiocytic proliferation with lymphophagocytosis (emperipolesis) characteristic of the Rosai Dorfman disease. The case and its management are discussed. PMID- 15805670 TI - Does increased 18FDG uptake reflect malignant transformation of a low-grade glioma? A diagnostic dilemma. AB - Benign gliomas of the brain show decreased uptake of 18F fluorodeoxyglucose (FDG) on positron emission tomography (PET). Malignant transformation is usually manifested by an increase of 18FDG uptake. A 45-year-old female has been followed up since 1987 by means of 18FDG-PET for a right hemispheric World Health Organization Grade II oligoastrocytoma. In 1996, increased epileptic activity was accompanied by increased 18FDG uptake within the temporal part of the tumor. After surgery, the epileptic seizures diminished. Histological examination of the resected tumor showed no change in the pathology when compared with the first biopsy. Localized temporal increase of 18FDG uptake was not associated with malignant progression. The decrease of seizure frequency might shed light on a putative connection of hyperglycolysis and epileptic discharges. PMID- 15805671 TI - Vertex extradural hematoma in association with Paget's disease of the skull. AB - Paget's disease of skull usually causes neurological complications such as basilar invagination, cranial neuropathies, etc. Occurrence of extradural hematoma in association with Paget's disease of skull is rare. A 48 year old man presented with headache and right upper limb weakness, two days after a fall from scooter. CT Scan of brain showed a large extradural hematoma at the vertex with the cranial vault showing features of Paget's disease. At surgery, no skull fracture or injury to the superior sagittal sinus was evident. There was diffuse oozing from the inner table of the skull, which showed features of Paget's disease. The extradural hematoma was evacuated and the patient made good recovery. He deteriorated a few hours after surgery. Follow up CT Scan showed diffuse brain swelling with minimal recollection of hematoma. Re-exploration showed a small recollection which was evacuated and the part of the oozing diseased skull was excised. The patient recovered completely. This is the first reported case of vertex extradural hematoma in association with the Paget's disease of skull. The unusual features are the absence of skull fracture and injury to the superior sagittal sinus. The increased vascularity of the skull due to Paget's disease has caused the hematoma by diffuse oozing from the inner table. PMID- 15805672 TI - Morgagni Stewart Morel syndrome--additional features. AB - A case of Morgagni Stewart Morel syndrome with progressive depression in frontal bone, headache, transient monoparesis, obesity; imbalance, neuropsychiatric symptoms and recurrent disc prolapse with absent right radial pulse is discussed. This syndrome was first mentioned 235 years back, but till now exact pathology is not known. Balance assessment using dynamic posturography was done, which revealed abnormal vestibular function. To our knowledge this is the first case examined for Dynamic Posturography. PMID- 15805673 TI - A case of sporadic Creutzfeldt Jakob disease with anterior visual pathway involvement. PMID- 15805674 TI - Vitamin B12 deficiency presenting with an acute reversible extrapyramidal syndrome. PMID- 15805675 TI - Safe surgical approach to deep pontomedullary cavernoma: an iMRI-assisted resection. PMID- 15805676 TI - Meningitis due to Escherichia vulneris. PMID- 15805677 TI - Extracranial redistribution causing rapid spontaneous resolution of acute subdural hematoma. PMID- 15805678 TI - Calcified vertebral artery and "dense basilar artery sign" in a patient with basilar territory infarction. PMID- 15805679 TI - Comparison of cognition abilities between groups of children with specific learning disability having average, bright normal and superior nonverbal intelligence. AB - BACKGROUND: Specific learning disabilities (SpLD) viz. dyslexia, dysgraphia and dyscalculia are an important cause of academic underachievement. AIMS: To assess whether cognition abilities vary in children with SpLD having different grades of nonverbal intelligence. SETTING: Government recognized clinic in a medical college. DESIGN: Cross-sectional study. SUBJECTS AND METHODS: Ninety-five children with SpLD (aged 9-14 years) were assessed. An academic achievement of two years below the actual grade placement on educational assessment with a Curriculum-Based test was considered diagnostic of SpLD. On basis of their nonverbal Intelligence Quotient (IQ) scores obtained on the Wechsler Intelligence Scale for Children test, the study children were divided into three groups: (i) average-nonverbal intelligence group (IQ 90-109), (ii) bright normal-nonverbal intelligence group (IQ 110-119), and (iii) superior-nonverbal intelligence group (IQ 120-129). A battery of 13 Cognition Function tests (CFTs) devised by Jnana Prabodhini's Institute of Psychology, Pune based on Guilford's Structure of Intellect Model was administered individually on each child in the four areas of information viz. figural, symbolic, semantic and behavioral. STATISTICAL ANALYSIS USED: The mean CFTs scores obtained in the four areas of information were calculated for each of the three groups and compared using one-way analysis of variance test. A P value < 0.05 was to be considered statistically significant. RESULTS: There were no statistically significant differences between their mean CFTs scores in any of the four areas of information. CONCLUSIONS: Cognition abilities are similar in children with SpLD having average, bright-normal and superior nonverbal intelligence. PMID- 15805680 TI - TNFR2 gene polymorphism in coronary artery disease. AB - BACKGROUND: Recently atherosclerosis and coronary artery disease (CAD) are considered to be inflammatory diseases. The genetic polymorphism in inflammatory markers has been well studied and found to be associated with development of CAD. AIM: To study the association of biallelic polymorphism at position 196 in exon 6 of tumor necrosis factor 2 (TNFR2) gene and coronary artery disease. SETTINGS AND DESIGN: The study design was a prospective case control study conducted at a tertiary referral center mainly catering to the north Indian population. MATERIALS AND METHODS: One hundred and fifty angiographically proven patients with coronary artery disease and one hundred and fifty age matched controls were genotyped for TNFR2 gene by polymerase chain reaction followed by analysis of restriction fragment length polymorphism. STATISTICAL ANALYSIS: Genotype frequencies were compared in patients and controls by Chi-square test. Binary logistic regression analysis was used to examine the relationship between genotypes and disease, incorporating other variables into the model. RESULTS: The incidence of CAD in those with MM genotype was 65% and in those with RM genotype was 42%. Genotype frequency shows significant association of MM genotype with development of CAD (P < 0.001; odds ratio-2.585; 95% confidence interval 1.533 4.359). The association of TNFR2 genotype with CAD persisted on logistic regression analysis. CONCLUSION: MM genotype of TNFR2 gene is associated with development of CAD and RM genotype appears to be protective. PMID- 15805681 TI - A case of autoimmune myopathy in pregnancy. AB - Autoimmune diseases are not found frequently with pregnancy in clinical practice. Polymyositis Dermatomyositis have a prevalence of 2.4-10.7/ 100,000 in general population. This is further low in pregnant women. It is associated with 57% perinatal morbidity and increased maternal and fetal mortality. Literature suggests that pregnancy outcomes are poorer if it manifests early in gestation while development or exacerbation in second or third trimester is associated with a better fetal prognosis. Not many case reports are published where the disease was diagnosed in third trimester. We present a case detected in third trimester, which was initially mistaken as a case of allergic reaction, however timely diagnosis and adequate management resulted in good fetal and maternal outcome. PMID- 15805682 TI - Acute viral E hepatitis with chronic liver disease (autoimmune hepatitis). AB - A 36 years old male presented with anorexia, jaundice and ascites. He was suffering from acute viral E hepatitis. In view of ascites, he was investigated for associated asymptomatic chronic liver disease (CLD). The CLD was diagnosed as cirrhosis with autoimmune hepatitis and was treated with steroid with good response. He is maintaining good health with low dose steroid, on follow up for 1 year. PMID- 15805683 TI - Maternal and perinatal outcome in pregnancy with hypothyroidism. PMID- 15805684 TI - Quadriparesis following wasp sting: an unusual reaction. PMID- 15805685 TI - Antibacterial resistance: current problems and possible solutions. AB - Antimicrobial resistance is a natural biological phenomenon of response of microbes to the selective pressure of an antimicrobial drug. Resistance may be inherent, which explains the phenomenon of opportunistic infection or acquired. Concern about the resistance increased in the late 1990's and since then, many governmental and agency reports have been published regarding the agricultural use of antibacterials, advising less use of antibacterials, appropriate choice of antibacterials and regimens, prevention of cross-infection and development of new antibacterials. The emergence of multidrug resistant strains of Gram-negative bacteria (Pseudomonas, Klebsiella, Enterobacter, Acinetobacter, Salmonella species) and Gram-positive organisms (Staphylococcus, Enterococcus, Streptococcus species) is the more worrisome in the present therapeutic scenario. Multidrug - resistant tuberculosis is another serious public health problems. Resistance to some agents can be overcome by modifying the dosage regimens (e.g., using high dose therapy) or inhibiting the resistance mechanism (e.g., beta-lactamase inhibitors), whereas other mechanisms of resistance can only be overcome by using an agent from a different class. It is urgently required to ban the sale of antibiotics without prescription, to use antibiotics more judiciously in hospitals by intensive teaching of the principles of the use of antibiotics and to establish better control measures for nosocomial infections. Thus, it is highly recommended that practicing physicians should become aware of the magnitude of existing problem of antibacterial resistance and help in fighting this deadly threat by rational prescribing. PMID- 15805686 TI - Tissue microarrays: potential in the Indian subcontinent. AB - Tissue microarrays (TMAs) are a means of combining hundreds of specimens of tissue on to a single slide for analysis simultaneously. The evolution of this technology to validate the results of cDNA microarrays has impacted tremendously in accurately identifying prognostic indicators significant in determining survival demographics for patients. TMAs can be generated from archival paraffin blocks, combined with sophisticated image analysis software for reading TMA immunohistochemistry, and a staggering amount of useful information can be generated in terms of the biomarkers useful in predicting patient outcome. There is a wide range of uses for the TMA technology including profiling of specific proteins in cancerous tissues and non-cancerous tissues. Given the wide variety of tissue resources available in India, investment in a dedicated TMA facility will be of immense use in the research arena in India. This review article discusses the basics of TMA construction, design, the software available for the analysis of this technology and its relevance to Indian scientists. A potential workflow structure for setting up a TMA facility is also included. PMID- 15805687 TI - Cancer preventive role of selected dietary factors. AB - Dietary behavior seems to be an important modifiable determinant for the risk of cancer. The evidences from several epidemiological studies suggest that higher intakes of fruits and vegetables have been associated with lower risk of cancer. Dietary phenolic and polyphenolic substances, terpenoids, dietary fibers, fish oils, some micronutrients present in foods of both plant and animal origin, and a reduction of caloric intake appear to inhibit the process of cancer development. Many dietary factors possess antioxidant and anti-inflammatory properties and cause induction of phase II enzymes like glutathione-S-transferases. It has been suggested that cruciferous vegetables play an important role in cancer prevention, and their chemopreventive effects are due to high glucosinolate content which under enzymatic hydrolysis produces bioactive compound isothiocyanates. Further, isothiocyanates of a wide variety of cruciferous vegetables are powerful inhibitors of carcinogenesis in experimental animal models. Several flavonoids present in fruits, tea, soya beans, etc. may be useful as cancer preventive agents. Similarly, ellagic acid, perillyl alcohol and resveratrol found in various fruits may have chemoprotective effect. Moreover, different vanilloids such as curcumin and gingerol have been shown to possess antioxidative properties. Nevertheless, in spite of several studies, still the effects of various ingredients are not clearly distinguished. In human, little convincing evidence has been established for the proposed protective effects of dietary constituents. It is an important future research goal to provide necessary evidences to support the chemopreventive role of different dietary factors, and also to clarify misunderstandings in this perplexing area. PMID- 15805688 TI - A study of histopathological features of medullary carcinoma of the thyroid: cases from a single institute in India. AB - BACKGROUND: The microscopic features of medullary carcinoma have been described in world literature, together with its behavior and molecular biology. However, no large study has been reported from India. AIMS: This study aims to analyse the clinical, and especially the pathological features of medullary carcinoma of the thyroid, and the surrounding thyroid. MATERIALS AND METHODS: In this study a total of 234 cases of medullary thyroid carcinoma (MTC) were gathered over a period of 3 decades. The clinical presentation, the microscopic features and the clinical outcome were analyzed. RESULTS: MTC was found to be twice as common in men as in women and for some reason it occurred 10 years earlier in women. The histology revealed certain interesting features like the presence of apoptosis in over half of the tumors, in addition to the other common and not so common histological findings (encapsulated variant, small cell variants, follicular pattern, rosettes, oncocytic change, osteosarcoma-like pattern, and cribriform pattern). The adjacent thyroid in about 19% of the cases showed optically clear nuclei in the follicles that were close to the tumor cells. These features were similar to those seen in papillary thyroid carcinoma. CONCLUSIONS: The thyroid adjacent to MTC showed nuclear changes, which are also found in papillary carcinoma of the thyroid. The occasional concurrent occurrence of these two tumors and the involvement of the RET gene in both medullary and papillary carcinomas, makes this observation worth discussing and studying further. PMID- 15805689 TI - Feasibility of breast conservation surgery in locally advanced breast cancer downstaged by neoadjuvant chemotherapy: a study in mastectomy specimens using simulation lumpectomy. AB - BACKGROUND: The response of locally advanced breast cancer (LABC) to neoadjuvant chemotherapy (NACT) offers these patients previously treated by mastectomy, the chance for breast conservation. AIM: This study aims to assess the feasibility of lumpectomy in patients with LABC treated by NACT, with residual tumor < or =5 cm. SETTINGS, DESIGN: Single group prospective study from August 2001 to June 2003 in a teaching hospital. MATERIALS AND METHODS: Thirty patients with LABC whose tumors reduced with NACT to 5 cm were included. Simulation lumpectomy was performed on the mastectomy specimens to achieve 1 to 2 cm clearance from tumor and hence margin negativity. Multiple sections of the inked margin were studied. STATISTICAL ANALYSIS: Margin positivity was correlated with patient factors. Chi square test and Fisher's exact test used as appropriate. P value 0.05 was considered significant. RESULTS AND CONCLUSIONS: After three cycles of NACT, 4 patients (13%) had complete clinical response including 2 with complete pathological response. Twenty-two (73%) showed partial response and 4, no response. Fourteen out of thirty (47%) had tumor involvement of margins. Tumors with post-chemotherapy size> 4 cm were margin positive in 10/13 (77%). Tumors with post-chemotherapy size>3 cm were margin positive in 13/24 (54%). Tumors with post-chemotherapy size 3 cm were margin negative in 5/6 (83%). Pre-chemotherapy tumor size and post-chemotherapy tumor size were significantly associated with margin positivity (P=0.003). Tumors in the subareolar location had significantly higher incidence of residual tumor in the nipple areola complex. (P=0.04). Margin positivity of lumpectomy on downstaged tumors can be reduced by removing the nipple areola complex in subareolar tumors and by limiting breast conservation to tumors with post-chemotherapy size < or =3 cm. PMID- 15805690 TI - Juvenile nasopharyngeal angiofibroma: a single institution study. AB - BACKGROUND: Juvenile nasopharyngeal angiofibroma (JNA) is a rare tumor of adolescent males and there is a paucity of Indian studies on this subject. AIMS: To present the experience of management of JNA at a single institution. SETTING AND DESIGN: This is a retrospective observational study of patients with JNA who presented at the Tata Memorial Hospital between May 1988 and August 2001. MATERIALS AND METHODS: Thirty-two patients with JNA were treated in the study period. Since the time period was prolonged and diagnostic and therapeutic protocols had undergone many changes, the patients were divided into two groups, namely 1988-1996 and 1997-2001. The age distribution, disease patterns, management approaches and treatment outcomes of patients in the two groups were recorded. Statistical analyses were done using students 't' test and test for proportion. RESULTS: The mean age at presentation was 16 years and more than 90% of the patients had Stage III or IV disease. Preoperative embolization was carried out in 19 patients. The surgical approaches used were median maxillectomy, infratemporal fossa, transpalatal, maxillary swing and craniofacial approach. The recurrence rate, complete resection rate and cure rates were 12.5%, 41% and 63% respectively. CONCLUSION: Surgery is the mainstay of treatment of JNA. Preoperative embolization and newer surgical approaches result in less blood loss and complete resection. Aggressive re-resection should be done for resectable recurrences reserving radiotherapy for unresectable, recurrent/ residual disease. PMID- 15805691 TI - Clinical features and prognostic factors of early breast cancer at a major cancer center in North India. AB - BACKGROUND: Data on the clinical profile of early breast cancer (EBC) from India is scant. Due to differences in genetics, environment, lifestyle, socio demographic structure and ethnicity, the presentation and behavior of breast cancer in India may be different. AIMS: To analyze the clinical presentation and outcome of EBC patients. SETTINGS AND DESIGN: A single center retrospective study. MATERIALS AND METHODS: Data from 487 EBC patients registered and treated at our institute from 1993 through 1999 were analyzed. Cox's multivariate regression test was used to determine prognostic factors for overall and disease free survival (OS & DFS). RESULTS: The median age was 47 years and 49.7% patients were pre-menopausal. Ninety-six per cent patients presented with a lump. Stages I, IIa, and IIb comprised 7.8%, 38.8%, and 47.6% respectively. Only 11.3% patients opted for breast-conserving surgery (BCS) while the remaining 88.7% underwent modified radical mastectomy (MRM). Adjuvant chemotherapy was administered to 275 (56.5%), and radiotherapy to 146 (29.9%). Estrogen receptor status was known in 173, of whom 93 (53.7%) were positive. Most patients were prescribed Tamoxifen for 5 years. At a median follow-up of 48 months, 126 (25.9%) patients had relapsed (systemic 107, loco-regional 19) and 94 (19.3%) had died. Five-year DFS and OS were 73% and 78%, respectively. On multivariate analysis, four positive nodes adversely influenced survival (P< 0.01). CONCLUSIONS: The median age at presentation was 47 years, significantly lower than most Western figures. The majority (86.4%) had a lump size > two cm. BCS was done in only 11% and the rest underwent MRM. Nodal involvement was the significant prognostic factor. PMID- 15805692 TI - Use of simple hematological, biochemical and clinical parameters to monitor response of multiple myeloma patients on high dose thalidomide therapy. AB - BACKGROUND: Evidence of increased bone marrow vascularity in multiple myeloma (MM) has led to the use of anti-angiogenic drugs especially thalidomide in relapsed or refractory patients. Currently, parameters such as serum/ urine electrophoresis for M (monoclonal) proteins, bone marrow biopsy with touch preparation and b2 microglobulin are routinely used to assess response to therapy. These investigations are expensive, invasive and require high technical setup. AIM: To correlate simple and routine hematological and biochemical parameters with the key marker of disease i.e. M proteins. SETTINGS AND DESIGN: This is an open label, uncontrolled, single-arm study. MATERIALS AND METHODS: Twenty nine refractory or relapsed multiple myeloma patients of both sexes (M=20, F=9) with age ranging between 35-72 years were initiated on 200 mg/day of thalidomide with fortnightly increments of 200 mg to a maximum tolerated dose not exceeding 800 mg/day. All hematological and biochemical parameters were monitored at monthly intervals for one year. STATISTICAL ANALYSIS: Correlation analysis was performed between hemoglobin (Hb), total leukocyte count (TLC), absolute neutrophil count (ANC), platelet count (PC), total proteins (TP), serum albumin and serum globulin on one hand and M protein levels on the other using Pearsons Correlation test by SPSS version 7.5. RESULT: Hb, TLC, ANC, PC and serum albumin levels showed a significant negative correlation with M proteins. A highly significant positive correlation existed between M proteins on one hand and TP and globulin levels on the other. Dryness of skin indicated positive response to therapy. These correlations were found to be significant at the end of one month of therapy in all the above-mentioned parameters except in TLC where it was significant after 2 months of thalidomide therapy. CONCLUSION: Results suggest that sustained efficacy of thalidomide therapy may be amenable to monitoring by these simple, inexpensive and easily available investigations after ascertaining an initial response by M protein and marrow plasmacytosis as these parameters closely follow M protein levels. However more studies are required to further substantiate these findings. PMID- 15805693 TI - The efficacy of postoperative radiation therapy in patients with carcinoma of the buccal mucosa and lower alveolus with positive surgical margins. AB - PURPOSE: A retrospective analysis to determine the efficacy of postoperative radiation therapy, in patients of carcinoma of the buccal mucosa and lower alveolus with pathologically verified positive surgical margins (PSM). MATERIALS AND METHODS: Ninety-four patients were analysed, who underwent surgery plus postoperative radiation therapy. Twenty-nine patients (31%) had PSM. Other pathological factors like nodal stage, number of nodes, bone involvement etc. were also analysed. RESULTS: Disease free survival (DFS) of patients with a PSM was significantly worse when compared with those with negative surgical margins (NSM). Poor DFS was also observed for variables like nodal stage, number of nodes and extranodal extension and radiation dose. In multivariate analysis only two variables showed significant impact on DFS, those were surgical margins and number of nodes. CONCLUSION: To conclude in our study median dose of 60 Gy in PSM patients was not able to improve DFS and showed poor results as compared with NSM patients. There is also evidence from other studies, to suggest that post operative radiation doses upto 60 Gy may not be sufficient to overcome this poor prognostic factor. To overcome this poor prognostic group patients, we in our institution are now employing radiation dose intensification and altered fractionation in an effort to improve our results. In physically fit patients we are trying to administer concomitant chemotherapy along with radiation treatment. PMID- 15805694 TI - Generalized neutrophilic dermatosis: a rare presentation of myelodysplastic syndrome. AB - We present a 30-year-old man admitted with generalized cutaneous lesions, fever and cough. Examination of skin biopsies of a papular lesion revealed dense neutrophilic infiltration of the upper dermis, so these lesions were diagnosed as neutrophilic dermatosis. Peripheral blood examination and bone marrow findings confirmed the diagnosis of myelodysplastic syndrome with excess blasts. The cutaneous lesions improved after administration of corticosteroid and follow-up bone marrow examination revealed a normocellular marrow. One year later he referred with acute myelogenous leukemia (AML-M0). Unfortunately, he did not respond to treatment and died a few months later due to disease progression. PMID- 15805695 TI - (18)FDG PET in primary oat cell carcinoma of the esophagus. AB - The role of FDG-PET in oat cell carcinoma of the esophagus is hitherto unexplored. A MEDLINE search using the terms "small cell carcinoma" or "oat cell carcinoma" combined with "FDG-PET" yielded no report on this issue till date. We report, in this article, two cases depicting the usefulness of this modality in the management of this uncommon neoplasm. While reevaluation of unsuspected metastatic sites missed by other modalities suggest its role in M staging, whole body FDG PET (both baseline as well as post treatment) may find important role in treatment monitoring and evaluation in residual viable disease, taking into account the systemic nature of the disease. PMID- 15805696 TI - Orthodox sometimes generates paradox. PMID- 15805697 TI - Quality of life assessment in patients with interstitial pulmonary fibrosis. PMID- 15805698 TI - Securing the safety and efficacy of macrolide therapy for chronic small airway diseases. PMID- 15805699 TI - Hepatocellular carcinoma in primary biliary cirrhosis at an early histologic stage: coincidental or causally related? PMID- 15805700 TI - Hypoglycemic attack: a rare triggering factor for takotsubo cardiomyopathy. PMID- 15805701 TI - Hyponatremia associated with selective serotonin reuptake inhibitors. PMID- 15805702 TI - Reversible ventricular dysfunction takotsubo (ampulla-shaped) cardiomyopathy. PMID- 15805703 TI - Hereditary periodic Fever syndromes in Japan. PMID- 15805704 TI - Chronic kidney disease as a metabolic syndrome with malnutrition--need for strict control of risk factors. AB - Patients with chronic kidney disease (CKD) have an increased risk for death from cardiovascular disease (CVD). They have multiple metabolic abnormalities that may accelerate atherosclerosis, such as hypertension, insulin resistance, and dyslipidemia, along with other CKD-related risk factors. In addition, a considerable proportion of patients with advanced stages of CKD are malnourished, presenting "metabolic syndrome with malnutrition". The presence of malnutrition/inflammation dramatically changes the apparent relationship between CVD death risk and some risk factors. For example, in stage 5 CKD patients on hemodialysis, a higher body mass index and a higher plasma cholesterol are predictors of better survival. To understand the paradoxic epidemiology, we should recognize risk factors for occurrence of CVD events and risk factors of fatality after an event. In this article, we review the unique situation of CKD, emphasizing the need of more strict control of both types of risk factors to improve survival of CKD patients. PMID- 15805705 TI - Risk of worsened encephalopathy after intravenous glycerol therapy in patients with adult-onset type II citrullinemia (CTLN2). AB - OBJECTIVE: We report a risk of worsening of encephalopathy by glycerol infusion when this osmotic agent is used for the treatment of brain edema in patients with adult-onset type II citrullinemia (CTLN2) caused by citrin deficiency. PATIENTS AND METHODS: We performed a retrospective investigation of 3 patients with CTLN2 treated for brain edema at our institute: a 31-year-old male patient and a 40 year-old female patient received treatment for encephalopathy-related brain edema with 10% glycerol infusion and 20% D-mannitol, and a 40-year-old male patient received only 20% D-mannitol infusion. In addition, we also performed a retrospective study in 11 CTLN2 patients reported previously (8 patients treated with 10% glycerol, 2 treated with 10% glycerol and 20% mannitol, and 1 treated with 20% mannitol). RESULTS: The 12 patients treated with 10% glycerol, including 2 of our patients, died due to rapidly deteriorating encephalopathy and brain edema. On the other hand, the 2 patients who received only 20% D-mannitol, including one of our patients, recovered with the disappearance of brain edema. CONCLUSION: In CTLN2 patients, glycerol infusion seems to be associated with exacerbation of encephalopathy itself and only mannitol should be used for the treatment of brain edema in patients with this disorder. PMID- 15805706 TI - Reassessment of the classification of the severity in idiopathic pulmonary fibrosis using SF-36 questionnaire. AB - OBJECTIVE: To investigate whether or not the newly revised classification of the severity of idiopathic interstitial pneumonia (IIP) is appropriate with respect to quality of life (QOL). METHODS: The association between the subscale of Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and pulmonary function or serum marker was analyzed using Pearson's correlation coefficient. The association between the subscale of SF-36 and the previous or newly revised classification of the severity of IIP was analyzed using Spearman's rank correlation test. PATIENTS: Forty patients with idiopathic pulmonary fibrosis (IPF) were enrolled. RESULTS: The mean deviation value scores for 7 items, excluding bodily pain (BP) in SF-36 were below the national reference values. % vital capacity (VC) was correlated with the 7 items excluding BP. However, neither serum LDH nor KL-6 values were correlated with any item in SF-36. According to the new or previous classification of the severity, severity was correlated with physical function, limitation of role functioning related physical problems and general health (GH); the correlation coefficient with the new one was slightly higher than the previous one. Based on these results, we established a unique draft on the classification of the severity. %VC <70% was added as an item for the newly revised classification in our draft. In our draft, there was rank correlation between the 7 items, excluding BP, in SF-36 and severity. CONCLUSION: With respect to QOL, the newly revised classification of the severity of IIP was not satisfactory, but the hypoxemia during exercise in patients with resting PaO(2) >80 Torr and reduction of VC were found to be important factors. PMID- 15805707 TI - Long-term macrolide antibiotic therapy in the treatment of chronic small airway disease clinically mimicking diffuse panbronchiolitis. AB - OBJECTIVE: In the current studies, we investigated the clinical effects of long term macrolide antibiotic therapy for patients with chronic small airway disease (CAD) that clinically and radiologically mimics but is pathologically distinct from diffuse panbronchiolitis (DPB). PATIENTS AND METHODS: Twenty-one Japanese patients were selected on the basis of clinical criteria for DPB and were categorized as DPB or CAD following histological evaluation of surgical lung biopsies. All patients received long-term macrolide therapy, and therapeutic results were compared for the DPB and CAD groups. RESULTS: Clinical, laboratory, radiological, and bacterial features, as well as neutrophilia in bronchoalveolar lavage fluid were strikingly similar in both groups. Long-term treatment with macrolides improved the clinical symptoms and PaO(2) in both groups. There was a significant improvement in forced expiratory volume in one second (FEV(1)), vital capacity (VC), and %VC in patients with DPB but not in patients with CAD. Neutrophilia in bronchoalveolar lavage fluid was also reduced following therapy in DPB patients but was refractory in CAD patients. CONCLUSION: Based on the different responses to macrolides, CAD might be associated with conditions distinct from those of DPB. Nevertheless, low-dose macrolide therapy may be applied in CAD to achieve clinical improvement, such as in respiratory symptoms and PaO(2). PMID- 15805708 TI - Hepatocellular carcinoma in a male patient with early stage (stage I) primary biliary cirrhosis. AB - The true incidence of hepatocellular carcinoma (HCC) in patients with primary biliary cirrhosis (PBC) remains undetermined due to limited epidemiological studies and some conflicting results. Some studies indicated that in PBC, male gender, cirrhosis, hepatitis C virus (HCV) superinfection, and history of blood transfusion are associated with the development of HCC, and the occurrence of HCC in the early stage of PBC is rare. We present herein a 75-year-old male patient with stage I PBC who developed oropharyngeal squamous cell carcinoma, followed by HCC and duodenal adenocarcinoma without hepatitis B or C virus infection. While it could be argued that the concurrence of HCC and stage I-PBC in our patient was coincidental, patients with early stage PBC should be strictly followed up as cirrhotic patients with PBC by monitoring the serum concentration of tumor markers for HCC and appropriate imaging methods. PMID- 15805709 TI - Hepatic encephalopathy due to intrahepatic portosystemic venous shunt successfully treated by interventional radiology. AB - We treated a 66-year-old woman with hepatic encephalopathy secondarily induced by an intrahepatic portosystemic venous shunt (IPSVS). In serial observations, the volume of the liver became smaller and encephalopathy could not be controlled with conservative therapy. We occluded the IPSVS successfully using percutaneous transcatheter embolization with micro coils. Following embolization, encephalopathy disappeared and blood flow of all branches of portal vein improved. In cases with an IPSVS without liver cirrhosis, blood flow in the portal vein and liver volume must be followed carefully, and interventional radiology may be considered effective in those who do not show a satisfactory response to conservative therapy. PMID- 15805710 TI - Chest pain and ST segment depression caused by expansion of gastric tube used for esophageal reconstruction. AB - We describe a 76-year-old man in whom a gastric tube was used for esophageal reconstruction via the anterior mediastinum after esophagectomy for esophageal cancer. Stenosis of the tube resulted in fluid accumulation, which directly compressed the heart and caused angina-like chest pain associated with ST-segment depression in lead V(2-3) on the electrocardiogram (ECG). Coronary angiography revealed no stenosis. Drainage of the fluid in the gastric tube resulted in immediate relief of symptoms and normalization of ECG. Angina-like chest pain associated with ST segment changes were caused by expansion of the gastric tube and compression of the heart. PMID- 15805711 TI - Pheochromocytoma presenting as recurrent hypotension and syncope. AB - We report a case with pheochromocytoma presenting as recurrent syncope due to hypotension. A 71-year-old man was admitted because of recurrent syncope and paroxysmal hypotension. He was diagnosed as having pheochromocytoma. In spite of a large volume of infusion and insulin therapy, syncope and paroxysmal hypotension continued. We speculated that hypotension was mainly due to vasodilatation caused by excess plasma epinephrine and prescribed a non-selective beta-adrenergic blocker. It stabilized blood pressure and syncope disappeared. The right adrenal tumor was excised and he was discharged in good condition. This case report provides some implications for the management of pheochromocytoma complicated with hypotension. PMID- 15805712 TI - Ampulla cardiomyopathy after hypoglycemia in three young female patients with anorexia nervosa. AB - Ampulla cardiomyopathy is named after the echocardiographic abnormalities occurring in this condition, characterized by extensive akinesis (ballooning ) of the apical region with hypercontraction of the basal segment of the ventricle. We describe 3 young female anorexia nervosa patients showing evidence of this cardiac complication after hypoglycemia. One case was complicated by echocardiographically confirmed ampulla cardiomyopathy while the other 2 patients showed increases in myocardial enzymes and transient electrocardiographic abnormalities consistent with this complication. The precipitating event for all three patients was hypoglycemic coma, and this is the first case report in which this factor lead to the complication of ampulla cardiomyopathy in anorexia nervosa patients. PMID- 15805713 TI - Severe starvation hypoglycemia and congestive heart failure induced by thyroid crisis, with accidentally induced severe liver dysfunction and disseminated intravascular coagulation. AB - A 69-year-old woman caught a cold resulting in nausea, vomiting, diarrhea and severe anorexia. Then she suffered progressively from dyspnea and leg edema, and finally became delirious. On admission severe hypoglycemia, hypothermia, marked tachycardia, generalized edema, mild jaundice and cachexy were noted. EKG showed atrial fibrillation. A chest X-ray, chest CT and echocardiography showed congestive heart failure. Therapeutic use of diuretics induced shock leading to serious liver dysfunction and disseminated intravascular coagulation. However, combined therapy by intravenous glucose, digitalis, diuretics, anti-fibrinolytic drug and hydrocortisone were effective. Addition of antithyroid therapy brought a further favorable outcome. PMID- 15805714 TI - Hyponatremia upon resumption of paroxetine therapy. AB - We present a 79-year-old woman with severe hyponatremia secondary to resumption of treatment with paroxetine, a selective serotonin-reuptake inhibitor antidepressant. Confusion and fatigue followed re-initiation of paroxetine after a 3-month hiatus. Hyponatremia, serum hypoosmolality, and urine hyperosmolality strongly suggested the syndrome of inappropriate secretion of antidiuretic hormone. Hyponatremia was quickly resolved after discontinuation of paroxetine and initiation of intravenous normal saline infusion together with oral fluid restriction. This case underscores the importance of monitoring serum sodium in elderly patients taking paroxetine, whether this represents a new prescription or reintroduction of the drug. PMID- 15805715 TI - Paradoxical peripheral embolism coincident with acute pulmonary thromboembolism. AB - Paradoxical embolism may occur in patients with acute pulmonary thromboembolism, when patent foramen ovale (PFO) coexists with pulmonary hypertension (right-left shunt). There have been few case reports of paradoxical embolism in peripheral arteries coincident with acute pulmonary thromboembolism. Here, we describe a case of paradoxical peripheral embolism associated with PFO complicated by acute pulmonary thromboembolism. The patient had severe peripheral ischemia due to a massive thrombus and was treated successfully by peripheral thrombectomy, thrombolysis, implantation of a permanent inferior vena cava filter and anticoagulation. PMID- 15805716 TI - Chronic necrotizing pulmonary aspergillosis complicated by a cavitary lesion caused by Pulmonary Mycobacterium-avium complex disease. AB - A 66-year-old woman who had undergone one year's treatment for pulmonary nontuberculous mycobacterial disease due to Mycobacterium avium (rifampicin, ethambutol, clarithromycin, streptomycin-->levofloxacin) five years earlier was admitted to our hospital because of continuous fever and a newly detected abnormal chest shadow, which was like a fungus ball in the right upper lobe on chest computed tomography in the giant cavitary lesion caused by pulmonary Mycobacterium-avium complex (MAC) disease. A diagnosis of chronic necrotizing pulmonary aspergillosis (CNPA) complicated by pulmonary MAC disease was made because Aspergillus niger was isolated from several sputum specimens, anti aspergillus antibody was positive, and clinical symptoms such as fever, were disclosed with the radiological finding of a fungus ball-like shadow and an infiltration shadow around the cavity. The patient had received various forms of antifungal chemotherapy, but the clinical effect had been poor. Since then, she had been slowly worsening. Although mycetomas, with the typical appearance of a fungus ball on a chest radiograph, have been reported to easily form in cavitary lesions caused by previous pulmonary tuberculosis, we believe, as illustrated by the present case, that they could also form in such lesions caused by pulmonary MAC disease, since the frequency of pulmonary nontuberculous mycobacterial disease has recently been increasing in comparison with that of pulmonary tuberculosis. PMID- 15805717 TI - Takotsubo (ampulla-shaped) cardiomyopathy associated with microscopic polyangiitis. AB - Recently, a cardiac disorder characterized by ballooning and hypokinesis at the apex has been described as takotsubo (ampulla-shaped) cardiomyopathy. We encountered a patient with a rare case of takotsubo cardiomyopathy associated with microscopic polyangiitis. A 70-year-old woman suddenly presented with ventricular dysfunction during the active phase of microscopic polyangiitis. The findings on echocardiograms and electrocardiograms were consistent with those of takotsubo cardiomyopathy. The ventricular dysfunction completely resolved after treatment with 40 mg/day of prednisolone and methylprednisolone pulse therapy. This unique type of cardiomyopathy can be a complication of microscopic polyangiitis. PMID- 15805718 TI - Takayasu myocarditis mediated by cytotoxic T lymphocytes. AB - A 15-year-old Japanese man was referred for evaluation of heart failure. Conventional heart failure therapy had little effect, and severe left ventricular dysfunction as well as elevated erythrocyte sedimentation rate persisted. Magnetic resonance angiography showed aortic dilatation with wall thickening characteristic of Takayasu's arteritis. An endomyocardial biopsy specimen revealed infiltration of natural killer cells and gamma delta T lymphocytes, which play major roles in vascular injury of Takayasu's arteritis. Prednisolone administration provided great benefits to cardiac function. These findings suggest that autoimmune cytotoxic mechanisms similar to those in arterial tissue may contribute to cardiac impairment in Takayasu's arteritis. PMID- 15805719 TI - A Japanese patient with familial Mediterranean fever associated with compound heterozygosity for pyrin variant E148Q/M694I. AB - Familial Mediterranean fever (FMF) is an inherited inflammatory disease occurring mainly in Mediterranean and Middle Eastern populations. FMF is caused by mutations in the MEFV gene that encodes pyrin/marenostrin. Here, we report a Japanese female FMF patient with heterozygosity for the compound pyrin E148Q/M694I showing recurrent fever, serositis or delay in skin wound healing. Her father and elder sister were heterozygous for pyrin variant M694I alone and sometimes suffered from mild fever or delay in wound healing, but her mother was heterozygous for pyrin variant E148Q alone and had no symptoms. This suggested that the inheritance of FMF occurred not only in an autosomal recessive manner but also in an autosomal dominant manner in this Japanese family, and the severity of the disease differed among the family members in relation to the mutation. In the treatment of FMF, colchicine, reserpine or prazosin hydrochloride have been reported to prevent the attacks, but, in our patient such drugs were ineffective or caused side effects, and only the anti-allergic drug azelastine was of benefit in relieving the attacks. PMID- 15805720 TI - Mycobacterium peregrinum infection in a patient with AIDS. AB - The patient, a 30-year-old housewife, visited a nearby doctor in mid August 2002 because of weight loss and neck swelling. HIV tests done at the hospital were positive. She was referred to and admitted to our hospital on October 2 for detailed examination and treatment of the neck tumor. A coat of epithelial debris extended from the oral cavity to the pharynx and an abscess and a fistula were found in the left tonsil. After hospitalization, an abscess culture revealed the presence of acid-fast bacteria, which was identified as Mycobacterium peregrinum. Treatment with imipenem and clarithromycin resulted in the normalization of CRP (0.1 mg/dl), on day 5 of treatment. The patient was discharged from the hospital after treatment for 2 weeks with imipenem and clarithromycin. Thereafter, the patient received continuous treatment with faropenem and clarithromycin for 4 more weeks, and has shown no signs of recurrence for 11 months to date. Only a few cases of infection with this bacterial strain have been reported. This infection is difficult to treat because most antituberculosis agents are not effective against it and there is limited availability of effective antibiotics. Medical treatment of infection caused by Mycobacterium peregrinum may be useful in such cases. PMID- 15805721 TI - Mesentery artery embolism and splenic infarction in infective endocarditis. PMID- 15805722 TI - Sacral arachnoid cyst associated with marfan syndrome. PMID- 15805723 TI - Pseudorabies virus propagated in rabbit kidney-derived RK13 cells is neutralized by natural IgM antibodies in normal swine serum which specifically lyse host cells. AB - Pseudorabies virus (PRV) propagated in rabbit kidney-derived RK-13 cells (PRV-RK) was neutralized by serum obtained from specific pathogen-free pigs through the activation of complement. The virus-neutralizing activity of swine serum was lost after treatment with ethylene glycol-bis-aminoethylether-N,N,N',N'-tetraacetic acid (EGTA) or ethylenediaminetetraacetic acid (EDTA). Anti-C1q and anti-IgM antibodies also inhibited virus-neutralizing activity. Though IgG-depleted swine serum neutralized PRV, IgM and IgG-free swine serum lost virus-neutralizing activity. Pre-incubation of swine serum with RK-13 cells, but not with swine kidney-derived CPK cells, at 4 degrees C eliminated the virus-neutralizing activity to PRV-RK. Results indicated that swine serum contained natural IgM against an antigen(s) on the RK-13 cell surface and that this surface antigen was integrated into the PRV envelope during the budding process. Thus the natural IgM in swine serum reacted with the RK-13 antigen on the viral envelope, activated the complement cascade and neutralized the PRV-RK. PMID- 15805724 TI - Mild hepatic fibrosis in cholesterol and sodium cholate diet-fed rats. AB - To date, the majority of research on hypercholesterolemia has focused on the effects of a high cholesterol diet on atherosclerosis and coronary heart disease. The toxic effects of cholesterol on the liver and the relationship between the intake of a high cholesterol diet and hepatic fibrosis, however, have not been investigated clearly or histopathologically. Male Wistar rats were fed a diet supplemented with 1.0% cholesterol and 0.3% sodium cholate for 12 weeks. Rats were sacrificed and analyzed via blood biochemistry, traditional microscopy and immunohistochemistry. Following the feeding of this diet, the rates of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase and total cholesterol in the rats were elevated consistently from week 3 and throughout the remainder of the experiment. From microscopic observation, hepatic necrosis, macrophage infiltration and steatosis increased markedly throughout the experiment. Hepatic fibrosis and myofibroblast proliferation were detected at weeks 9 and 12. Mast cell appearance was proportional to the degree of hepatic damage. These findings suggest that hepatic fibrosis is inducible by a high cholesterol diet and is likely the result of the interaction between several different cell types (i.e., macrophages, myofibroblasts, and mast cells) in an inflammatory milieu. Hypercholesterolemia should be considered as a risk factor for hepatic fibrosis as well as atherosclerosis and coronary artery disease. PMID- 15805725 TI - Bovine paraoxonase 1 activities in serum and distribution in lipoproteins. AB - Paraoxonase 1 (PON1) is an enzyme associated with high-density lipoprotein (HDL) and has a protective effect against oxidation of lipoproteins in mammals. We investigated PON1 enzyme activities in bovine serum and its distribution among bovine serum lipoproteins. Paraoxonase activity and arylesterase activity in serum (152 Holstein cows and 42 Japanese Blacks) were 275 +/- 55 U/ml and 130 +/- 27 U/ml (mean +/- SD), respectively. There was a high correlation (r=0.962) between the two enzyme activities, and the activity ratio of paraoxonase/arylesterase did not exhibit individual variation. More than 85% of both paraoxonase and arylesterase activities were detected in the HDL fraction separated by ultracentrifugation. The 43-kDa protein in the HDL fraction was identified as bovine PON1 by N-terminal amino acid sequence analysis. Bovine PON1 was purified by ultracentrifugation and preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis, and an anti-bovine PON1 antiserum was developed. The concentration of PON1 protein determined by immunoblotting was closely correlated (r=0.976) with paraoxonase activity in serum. Bovine HDL was further fractionated into subpopulations, and the distribution of PON1 was examined. Paraoxonase activity and PON1 protein increased with decreasing HDL size and approximately 60% of total paraoxonase activity was distributed in the heavy HDL fraction. The different distributions of PON1 among HDL subpopulations might be concerned to the function and metabolism of bovine HDL. PMID- 15805726 TI - Enzyme-linked immunosorbent assay for the detection of canine Leptospira antibodies using recombinant OmpL1 protein. AB - OmpL1 is a 31-kDa outer membrane protein characterized in 1993 and known to be expressed only in pathogenic Leptospira spp. Recombinant OmpL1 (GST-rOmpL1) was expressed for use as an ELISA antigen for the detection of anti-Leptospira antibodies. In immunoblot analysis, the protein reacted with sera of dogs infected with three different serotypes of Leptospira interrogans, while did not react with sera of dogs both uninfected negative controls and infected with Borrelia burgdorferi, which is closely related to Leptospira spp. Moreover, in ELISA using GST-rOmpL1, the optical density (O.D.) values from the positive controls were very high (1.125 +/- 0.549). In contrast, the O.D. values from clinically healthy dogs and dogs with diseases other than leptospirosis were very low (0.109 +/- 0.046 and 0.089 +/- 0.046, respectively). These data suggest that the detection of anti-Leptospira antibodies by ELISA using the GST-rOmpL1 protein can be applied for diagnosis of canine leptospirosis. PMID- 15805727 TI - Differentiation between bovine and ovine strains of Histophilus somni based on the sequences of 16S rDNA and rpoB gene. AB - Nucleotide sequences of 16S rDNA and rpoB gene of 25 bovine and 6 ovine Histophilus somni strains were determined to detect subtle differences between the host animal species. The 1465 nucleotide residues of the 16S rDNA exhibited levels of sequence similarities of 99.4% or more. The high sequence similarity of the 16S rDNA of recently described species H. somni was confirmed in the 31 strains from cattle and sheep. These results suggested that the intra-specific diversity of 16S rDNA was limited in bovine and ovine strains of H. somni. The specific association of strains was also observed in the 311 bp region of rpoB gene which sequence similarities were 98.6% or more. However, the phylogenetic tree analysis of the rpoB gene showed that the ovine strains appeared to form a subgroup recovered in 70% of the bootstrap trees. In the 311 bp region of the ovine strains, a HincII restriction endonuclease site was detected. The PCR amplified rpoB DNA of 46 bovine and 20 ovine H. somni strains were examined for the digestion with HincII. As the results, 17 strains of ovine strains were cleaved by the enzyme but none of the bovine strains appeared to possess the restriction site. The restriction enzyme analysis of rpoB gene may be useful to differentiate ovine strains from bovine strains of H. somni. PMID- 15805728 TI - Isolation and characterization of staphylococci from external auditory meatus of dogs with or without otitis externa with special reference to Staphylococcus schleiferi subsp. coagulans isolates. AB - Staphylococci were isolated from the external auditory meatus in 14 (48.3%) of 29 dogs affected with otitis externa (OE dogs) and 28 (68.3%) of 41 dogs without OE (non-OE dogs). Twenty-two OE isolates were identified as belonging to 12 species, and 42 non-OE isolates were identified as belonging to 13 species. The predominant species found in both OE and non-OE isolates were S. intermedius, and S. epidermidis. Thirty-eight (59.4%) of 64 isolates were resistant to one or more of the 17 antimicrobial agents tested. Resistance to PCG and ABPC was most frequent. S. schleiferi subsp. coagulans, a recent etiologic agent of canine OE, was isolated from OE and non-OE dogs. All of the 5 S. schleiferi subsp. coagulans isolates showed typical characteristics. No clear difference in the extracellular enzyme or toxin profiles, nor in the PFGE patterns, was demonstrated between the OE and non-OE isolates of S. schleiferi subsp. coagulans. A new PCR primer set specific for 16S rDNA was designed to identify strains of S. schleiferi subsp. coagulans. The amplified fragment was detected in all of the 5 isolates as well as the type strain GA 211 (=JCM 7470) and a reference strain GA 11, but was not detected in any strains of the related species, S. aureus, S. intermedius and S. hyicus. The PCR may allow a simple, rapid and precise identification of S. schleiferi subsp. coagulans, in addition to the standard tube test for free coagulase. PMID- 15805729 TI - Prevalence and characterization of Staphylococcus aureus and enterotoxigenic Staphylococcus aureus in retail raw chicken meat throughout Japan. AB - A total of 444 samples of raw chicken meat (thighs, breasts, wings, livers, gizzards, hearts and ovaries) that retailed at 145 different supermarkets in 47 prefectures in Japan were examined for contamination with Staphylococcus aureus in association with its enterotoxigenicity. S. aureus was isolated from 292 (65.8%) of the samples, and from 131 of the 145 supermarkets. There was no significant difference in the detection rate of S. aureus according to the type of meat examined. About 80% of 714 isolates belonged to the poultry (57.1%) and human biotypes (22.1%). Seventy-eight (21.7%) of 360 isolates were enterotoxigenic and isolated from 78 samples in 53 supermarkets in 31 prefectures. Staphylococcal enterotoxins (SEs) produced were SEB (50 isolates), SEA (14), SEC (8), SED (2), SEA+SEB (2), and SEA+SEC (2). Most of the enterotoxigenic isolates belonged to the human and poultry biotypes, coagulase type VII, VIII or IV, and were lysed by phages of group III. Identical SE types, biotypes, coagulase types and pulsed-field gel electrophoresis (PFGE) patterns were shown in isolates from different types of meat at the same supermarket and from samples taken from different supermarkets in the same prefectures or in isolates from samples obtained from several different prefectures. Among the 50 SEB-producing isolates, 27 yielded three similar PFGE patterns that differed by only a few fragments, suggesting that they were closely related genetically. The three patterns were found in isolates of samples that retailed at 17 supermarkets in 11 prefectures, indicating that they may be disseminated among raw chicken meat in Japan. PMID- 15805730 TI - GnRH treatment at CIDR insertion influences ovarian follicular dynamics in Japanese black cows. AB - Ovarian follicular dynamics and estrous synchronization after Gonadotropin releasing hormone (GnRH) treatment at Controlled Internal Drug Releasing device (CIDR) insertion were investigated in Japanese Black cows. CIDR was inserted for eight cows at 7 days after estrus. Cows were allocated to either Group A: 8-day CIDR insertion with GnRH treatment on d 0 (n=4, d 0=CIDR insertion) or Group B: 8 day CIDR insertion (n=4). Both groups were injected with prostaglandin F2alpha (PGF2alpha) on d 7. Ultrasonography and blood sampling were performed twice daily. Intensive sampling was performed every 15 min for 8 hr to determine the pulsatile release of LH on d -1, d 5 and d 10. Three of four cows showed intermediate ovulation within 2 days after GnRH treatment during CIDR insertion in Group A, whereas no ovulation was found in Group B. Three of four cows in Group A and all four cows in Group B ovulated after CIDR removal. Plasma progesterone concentrations from d 3 to d 7 in three intermediate ovulatory cows in Group A (8.4 +/- 1.6 ng/ml) was significantly higher than those in Group B (4.1 +/- 1.2 ng/ml; 4 cows) during CIDR insertion (P<0.01). Interval to estrus and ovulation after CIDR removal was observed at 60.0 +/- 12.0 hr and 76.0 +/- 6.9 hr in three cows in Group A, and 75.0 +/- 15.1 hr and 93.0 +/- 20.5 hr in Group B, respectively. There was a significant increase in LH pulse frequency on d 10 compared on d -1 or d 5 in both groups (P<0.05), in addition those on d 10 in Group A tended to be higher than in Group B. As a result, GnRH treatment at CIDR insertion at 7 days after estrus induced intermediate ovulation with formation of corpus luteum (CL) and rather synchronized emergence of ovulatory follicle during CIDR insertion. These induced CL increased plasma progesterone concentrations and contributed to precise synchronization. PMID- 15805731 TI - Acoustic feature of barks of ovariohysterectomized and intact German Shepherd bitches. AB - This study was carried out to investigate the effect of ovariohysterectomy on vocalization during territorial aggression of German Shepherd dogs. Sixteen clinically healthy dogs of 5 to 10 months old were assigned randomly to one of two groups: ovariohysterectomy or control. Their behaviors and vocalizations induced by the approach of a strange with a strange dog were recorded using digital camcorder at four and five months after surgery and were analysed. When territorial aggression was induced, dogs in ovariohysterectomized group showed more offensive territorial aggression. The bark was the most frequent vocalization. In this study, the average number of barks was 45 times in ovariohysterectomized group, and 26 times in the control group. The pitch of vocalization was significantly lower in ovariohysterectomized group than control group. First formant, second formant, third formant, and fourth formant frequency in ovariohysterectomized group, which represent the degree of sound energy in specific frequency, were lower than those of control group. Ovariohysterectomy of bitches may influence the frequency of aggressive vocalization and affect the acoustic feature of dogs' vocalization. Analysis of vocalization could be a useful method of evaluating the dogs' intention. PMID- 15805732 TI - Loss of milk yield due to Akabane disease in dairy cows. AB - Akabane disease is an infection with clinical signs of congenital malformation and abortion in ruminants. Abnormal parturitions caused by Akabane disease result in great economic loss. The purpose of this study is to estimate the reduction in the milk yield from abnormal parturition due to Akabane disease. The data were collected from 33 Holstein cows on 11 farms. The animals had abnormal parturitions during the period from September 1998 to March 1999, and were diagnosed as having Akabane disease. The mean and standard deviation of the rate of reduction in the milk yield of 33 cows after abnormal parturition caused by Akabane disease was -11.4 +/- 14.9%. The means and standard deviations of the rate of reduction of four cows calving abnormally at 220-239 days of gestation, nine cows calving abnormally at 240-270 days of gestation, and 20 cows calving abnormally at 271-300 days of gestation were -26.6 +/- 24.7%, -14.7 +/- 11.0%, and -6.9 +/- 12.3%, respectively. In this study, we demonstrated that the rate of reduction in the milk yield in cows affected by Akabane disease was -11.4 +/- 14.9%, but values as high as -26.6 +/- 24.7% were reached in the comparison with the milk yield obtained after normal parturition. PMID- 15805733 TI - Role of gap junctional intercellular communication (GJIC) through p38 and ERK1/2 pathway in the differentiation of rat neuronal stem cells. AB - Gap junctional intercellular communications (GJIC) contributes to neural function in development and differentiation of CNS. In this study, we have investigated the expression of GJIC during the differentiation of neuronal stem cells and 12-O tetradecanoylphorbol-13-acetate (TPA)-induced neuronal stem cell-derived cells from rat brain. During neuronal stem cell differentiation, expressions of Cx43 and 32 were increased for the duration of 72 hr, however the effect were decreased on the 7d. In the neuronal stem cell-derived cells, pretreatments with p38 MAP kinase inhibitor, SB203580, and MEK inhibitor, PD98059, could protect GJIC against TPA-induced inhibition of GJIC. Our data suggest that GJIC plays an important role during neuronal stem cell differentiation, and ERK1/2 and p38 MAP kinase signaling pathway may be closely related functionally to regulate gap junction in rat neuronal stem cell-derived cells. PMID- 15805734 TI - Blood parasites of some West African rainforest birds. AB - A total of 969 birds representing 121 species of 21 families from the West African nations of Cameroon, Equatorial Guinea and Ivory Coast were examined for haematozoa using thin blood smears; 277 individuals (28.6%) harbored blood parasites. The parasites identified included species of Haemoproteus (7.7% prevalence), Plasmodium (10.7%), Leucocytozoon (4.6%), and Trypanosoma (7.3%). In addition, microfilariae of filariid nematodes were present in 3.6% of the individuals examined. The birds were collected over a period of 12 years, from 1989-2001, from rainforest and ecotone habitats. We report a relatively high prevalence of parasites in colonial nesting birds, and two species of ground nesting birds. In addition, we compared data from bird species collected at a site identical to a previously published study, and did not find significant differences in parasite prevalence between the two years constituting two different seasons. Our results are also compared to other studies in Africa that implement similar and different methodologies. PMID- 15805735 TI - Hypothalamo-pituitary-adrenal gland axis in mice inhaling toluene prior to low level long-term exposure to formaldehyde. AB - We studied the change in the hypothalamo-pituitary-adrenal gland (HPA) axis upon adding prior toluene inhalation to our previous formaldehyde inhalation experiments to determine whether short term exposure to relatively high levels of toluene triggers multiple chemical sensitivity (MCS). Data come from immunocytochemical, morphometrical and RT-PCR measurements. Four groups of adult female mice were exposed to differing concentrations (0, 80, 400, and 2,000 ppb) of formaldehyde for 16 hr/day, 5 days/week for twelve weeks, after the mice were exposed intranasally to 500 ppm toluene per mouse for 6 hr/day, for 3 days. We found that the number of corticotropin releasing hormone (CRH)-immunoreactive (ir) neurons was up-regulated according to the amount of formaldehyde as well as inhalation of formaldehyde alone in our previous experiment. The proportion of adrenocorticotropin hormone (ACTH)-ir cells increased according to the formaldehyde concentration, though there was no significant difference between the 400 and 2,000 groups. The number of ACTH-ir cells was higher in the 400 group than in the other groups (0, 80, and 2,000). Expression of ACTH-mRNA was also up regulated according to the quantity of formaldehyde. The sinusoid in the anterior pituitary showed more dilatation in the 400 and 2,000 groups than in the control group, especially in the 2,000 group. We propose that exposure to toluene prior to inhalation of formaldehyde has no effect on the HPA axis and as a trigger of MCS, although greater sinusoid dilatation was found in the anterior pituitary gland at higher concentrations of formaldehyde. PMID- 15805736 TI - Murine goblet cell hypoplasia during Eimeria pragensis infection is ameliorated by clindamycin treatment. AB - The goblet cell (GC) and the intestinal mucus are important in preventing invasion of the mucosa by luminal microorgansisms. GC responses in the jejenum, cecum and colon of C57BL/6 mice during the course of infection with the large intestinal-tropic coccidian, Eimeria pragensis (E. pragensis), were investigated histologically. The numbers of large intestinal GCs (cecum and colon) gradually decreased (hypoplasia) in association with development of endogenous stages of parasite life cycle. The effect was transient and recovery of GC numbers was associated with resolution of coccidial infection. The jejunal GC numbers were not affected by E. pragensis infection. Clindamycin treatment in the infected mice reduced numbers of intracellular parasites and significantly increased the numbers of large intestinal GCs compared with untreated, infected mice. PMID- 15805737 TI - First demonstration of bovine herpesvirus 2 infection among cattle by neutralization test in Japan. AB - Seroepidemiological surveys were performed on neutralizing antibody to bovine herpesvirus 2 (BoHV-2) among cattle in Japan. A total of 1,819 sera were collected from cattle in 27 prefectures from 1997 to 1998. Antibodies were detected in only 18 sera collected from 4 prefectures. However, the most prevalent areas of the infection were found in two islands located in the subtropical zone. Additional 353 sera were collected in three including these islands in 1999-2001. The antibody-positive rates in the farms in these islands ranged from 10% to 81.1%. It was confirmed that BoHV-2 was prevalent in these areas. However, the infection seemed to be latent, because no diseases have been noticed. This is the first report showing the presence of BoHV-2 infection among cattle in Japan. PMID- 15805738 TI - Polymorphisms of caprine PrP gene detected in Japan. AB - Polymorphism of the PrP gene is a primary factor influencing susceptibility and incubation period in natural and experimental scrapie in sheep and goats. Polymorphisms of the caprine PrP gene in Japan were examined in 118 goats. Eight allelic variants and 19 genotypes were obtained. Amino acid polymorphisms were observed at 7 codons: 102, 142, 143, 240, 127, 146 and 211 (the latter 3 are novel polymorphisms). The polymorphisms at codons 142M and 143R, which are associated with the resistance to scrapie, were relatively rare in the present study. Thus, the present results provide information about the caprine PrP gene that may be useful for assessing the risk of goat scrapie. PMID- 15805739 TI - Nucleocapsid-like structures of Ebola virus reconstructed using electron tomography. AB - Electron tomography (ET) is a new technique for high resolution, three dimensional (3D) reconstruction of pleiomorphic macromolecular complexes, such as virus components. By employing this technique, we resolved the 3D structure of Ebola virus nucleocapsid-like (NC-like) structures in the cytoplasm of cells expressing NP, VP24, and VP35: the minimum components required to form these NC like structures. Reconstruction of these tubular NC-like structures of Ebola virus showed them to be composed of left-handed helices spaced at short intervals, which is structurally consistent with other non-segmented negative strand RNA viruses. PMID- 15805740 TI - Perirenal pyelocaliceal diverticulum in an infant cat. AB - A 3-month-old female tortoise-shell cat showing azotemia died with a marked swollen abdomen. Necropsy revealed a huge perirenal cyst (8.5 x 6.0 x 4.5 cm) on the ventral aspect of the right kidney. The cyst was filled with the pellucid yellow fluid with a smell of urine. The lumen was connected with irregularly dilated renal pelvis by a narrow channel passing through the renal parenchyma. The cyst was lined by epithelial cells and its wall was consisted of collagen fibers and smooth muscle cells as that of the renal pelvis and ureter. Renal parenchyma adjacent to the channel showed interstitial infiltration of the lymphoid cells. The cyst was a diverticulum of the renal pelvis due to an impaired development. PMID- 15805741 TI - Susceptibility of Chinese hamsters (Cricetulus griseus) to the infection of Babesia microti. AB - Chinese hamsters were examined for the susceptibility to the infection with Babesia microti based on the hematological parameters during the course of infection. A marked decrease in the RBC count, Ht value, Hb concentration, and an increase in WBC count due to the development of neutrophils or monocytes were recognized with the progress of parasitemia. Remarkable clinical findings were anemia and persistent infection with a low level of parasite burden in the chronic and convalescent stages. From these findings, it was concluded that Chinese hamsters were susceptible to infection with B. microti and would be useful for infection examination with the parasite. PMID- 15805742 TI - Identification of genotypes of Giardia intestinalis isolates from a human and calf in Japan. AB - Giardia intestinalis is recognized as a significant pathogen in humans and animals, causing diarrhea. Recent molecular studies indicate that G. intestinalis is composed of genetically distinct multiple genotypes. Therefore, it is valuable to distinguish among genotypes in the epidemiology of Giardia infection in humans and animals. Although G. intestinalis has been found in humans and animals in Japan, the genotype of isolates remains unclear except for several isolates from dogs, because identification has been performed only by conventional microscopy. We report herein the genotypes of G. intestinalis isolates distinguished by a phylogenetic analysis. G. intestinalis isolates originated from a patient and a calf were found to have Assemblage B and E, respectively. PMID- 15805743 TI - Hyperlipemia of captive bottlenose dolphins during pregnancy. AB - In this study values for total cholesterol and triglycerides were measured in 110 blood samples taken from 360 days pre-partum to 90 days post-partum in ten parturitions of six bottlenose dolphins, and in 75 blood samples when the dolphins were not pregnant as a control group. The average total cholesterol values in the second, third and fourth stages and in the puerperium were significantly higher than the average value of the control group by 11.0%, 30.2%, 19.3% and 13.4% respectively. The average triglycerides values for the third and fourth stages and in the puerperium were also significantly higher than those in the control group by 59.7%, 84.3%, and 42.1% respectively. PMID- 15805744 TI - Clinicopathological survey of 101 canine mammary gland tumors: differences between small-breed dogs and others. AB - Clinicopathological features of mammary gland tumors (MGTs) in 101 dogs were evaluated retrospectively. The incidence of histological malignancy in 60 small- and 41 other-breed dogs were 25% and 58.5%, respectively. In 82 epithelial MGTs, small-sized tumors (< 3 cm) or non-invasive tumors were predominant in small breeds. In multivariate survival analysis, small breed (p=0.048) and lower stage of tumor cell invasion (p=0.006) were significantly associated with longer survival time. These results suggest that the incidence of histological or biological malignancy in MGTs is lower in small-breed dogs than in others. PMID- 15805745 TI - Surveillance of chronic wasting disease in sika deer, Cervus nippon, from Tokachi district in Hokkaido. AB - Surveillance of chronic wasting disease (CWD) was conducted by performing Western blot analysis of tissue samples from 136 sika deer (Cervus nippon) killed by hunters in the Tokachi district of Hokkaido Island. No prion protein (PrPSc) associated with CWD was detected in any of the samples. To assess amino acid polymorphisms of the sika deer PrP gene, nucleotide sequencing of the PrP gene was performed. The only amino acid polymorphisms detected were 3 silent mutations at nucleotide positions 63, 225 and 408. These results suggest that sika deer in the Tokachi district are genetically homogeneous, and are not infected with CWD. PMID- 15805746 TI - Oral leiomyosarcoma in a woodchuck (Marmota monax). AB - We report the case of a 4-year-old female woodchuck (Marmota monax) which presented with a white, firm and discrete mass in the hard palate. The mass extended into the oral cavity but it was well separated from the surrounding tissues. Histology of the tumor showed a malignant mesenchymal tumor with pleomorphic spindle cells varying in degrees of differentiation and density. The neoplastic cells had moderate amounts of granular or fibrillar eosinophilic cytoplasm with indistinct cell margins. Nuclei were oval to elongated and frequently blunt-ended with vesicular chromatin. Immunohistochemical study showed that the neoplastic cells expressed vimentin and alpha-smooth muscle actin but did not express desmin, pan-cytokeratin, and S-100. Therefore, histology and immunohistochemistry revealed that the tumor was oral leiomyosarcoma. Oral cavity is an extremely rare site for leiomyosarcoma and the present case is the first report of spontaneous oral leiomyosarcoma in animals. PMID- 15805747 TI - [Evaluation of the surgical results of Achilles tendon ruptures by gait analysis and isokinetic muscle strength measurements]. AB - OBJECTIVES: We evaluated the long-term results of surgical treatment of Achilles tendon ruptures. METHODS: Fifteen male patients (mean age 39.5 years; range 28 to 58 years) underwent surgery for Achilles tendon ruptures. All the ruptures but one occurred during sport recreation. Eleven patients were treated within the first week of trauma. After surgery, a cast was applied for four weeks, after which it was switched to a polyethylene splint that did not allow dorsiflexion. Partial weight bearing was allowed in the fourth postoperative week. Subjective and objective evaluations were made according to the system by Thermann et al. Muscle strength was measured by an isokinetic dynamometer and gait analysis was performed. Data from the healthy sides were used for comparisons. The mean follow up was 16.8 months (range 8 to 48 months). RESULTS: Seven patients (46.7%) had no pain, while six patients (40%), one patient (6.7%), and one patient complained of pain during heavy, medium, and light sports activities, respectively. Muscle strength did not decrease in nine patients (60%). Seven patients (46.7%) returned to preinjury activity levels. Twelve patients (80%) evaluated the outcome as very good or good. The mean muscle atrophy on the affected side was 0.43 cm. The overall results were very good or good in nine patients (60%), moderate in four patients (26.7%), and poor in two patients (13.3%). One patient had superficial infection and delayed healing. Isokinetic measurements showed decreased peak torque, and increased total work. By gait analysis, no significant relationship was found between the dorsiflexion and plantar flexion difference and forefoot ground reaction forces. CONCLUSION: Early surgical treatment of Achilles tendon ruptures followed by a shorter immobilization period is associated with satisfactory results, and in the long-term, with a higher rate of patient satisfaction. PMID- 15805748 TI - [Treatment of femoral shaft fractures and pseudoarthrosis with compressive and interlocking intramedullary nailing]. AB - OBJECTIVES: We evaluated the results of compressive and interlocking intramedullary nailing in femoral shaft fractures. METHODS: Thirty-three femoral fractures of 30 patients (23 males, 7 females; mean age 36.9 years; range 14 to 80 years) were treated by compressive and interlocking intramedullary nailing. There were 26 closed and seven open (2 type II, 5 type IIIA) fractures. In three femurs, intramedullary nailing was performed due to pseudoarthrosis. According to the Winquist classification, there were 13, 8, 8, and 4 type I to IV fractures, respectively. Clinical, radiographic, and functional results were evaluated according to the Thoresen criteria. The mean follow-up was 29 months (range 6 to 29 months). RESULTS: The mean operation time was 105.7 minutes. Union was obtained in all the patients within a mean duration of 19.6 weeks. In six cases, dynamization was performed due to insufficient callus formation. The results were excellent in 17 patients (56.7%), good in eight patients (26.7%), fair in three patients (10%), and poor in two patients (6.7%). Complications included shortening (n=3; 2 to 4 cm), varus deformity (n=2; 8 and 11 degrees), external rotation (n=3), restriction in knee flexion (n=3), superficial infection (n=8), trochanteric bursitis (n=3), and irritation of the skin by distal screws (n=2). CONCLUSION: High rates of union with low complication rates makes interlocking intramedullary nailing an appropriate method in the treatment of femoral shaft fractures in adults. Compression with top screws contributes to the stability of fixation by eliminating any looseness through the fracture line. PMID- 15805749 TI - [Surgical management of hip instabilities in children with spina bifida]. AB - OBJECTIVES: We evaluated the results of surgical management of hip instability in children with spina bifida (SB). METHODS: Twenty-eight hips of 26 patients (16 girls, 10 boys; mean age 4.5 years; range 3 to 6 years) were surgically managed for hip instability (subluxation/dislocation) associated with SB. Twenty-four patients (2 bilateral dislocations) had low-level lesions (L4-sacral) and a potential to walk, of which 16 patients presented with unilateral dislocation with functional problems including significant (>2 cm) limb-length discrepancy and scoliosis. Two patients had high-level lesions (thoracic-L3) associated with unilateral dislocations and were unable to walk. Those with a high-level lesion and some patients (9/26 hips) with a low-level lesion also had hip flexion contractures. Treatment included open reduction, pelvic osteotomy, proximal femoral osteotomy when necessary, and a spica cast. The patients were clinically and radiographically monitored for a mean of 38 months (range 30 to 48 months). RESULTS: Of 16 patients with functional problems, 14 patients had improvement in their gait patterns, while limb-length discrepancy and scoliosis persisted in two. The remaining 10 patients maintained their preoperative functional statuses. The mean range of motion of the hips decreased postoperatively; however, none of them developed joint stiffness. Early postoperative complications included superficial wound infections in three patients, and distal femoral diaphyseal fractures in two patients. Three patients required removal of the implants in the sixth month due to subcutaneous prominence thereof. Late radiographs of three patients showed recurrent subluxations, which did not require any intervention. CONCLUSION: Although surgical treatment of hip problems associated with high level lesions may be unrewarding in children with SB, those associated with low level lesions can be successfully managed with proper surgical indications. PMID- 15805750 TI - [Closed reduction and percutaneous pinning with three Kirschner wires in children with type III displaced supracondylar fractures of the humerus]. AB - OBJECTIVES: We evaluated the results of closed reduction and percutaneous pinning in the treatment of displaced supracondylar fractures of the humerus in children. METHODS: Sixty-one children (41 boys, 20 girls; mean age 7.6 years; range 2 to 13 years) were treated for type III displaced supracondylar fractures of the humerus within the first six hours after trauma. Initial neurologic examinations were normal. Treatment included closed reduction followed by percutaneous pinning with two lateral and one medial K-wires, the last being placed during decreased (80 to 90 degrees) elbow flexion. The ulnar nerve was examined intraoperatively, at the time of discharge, at the end of two weeks, and in the third and six months. On final examinations, the range of motion and carrying angles of both elbows were measured with a goniometer and anteroposterior and lateral radiograms of the affected elbows were examined. The results were evaluated using the clinical scoring system developed by Flynn et al. The mean hospital stay was 3.4 days (range 1 to 10 days) and the mean follow-up was 28.9 months (range 17 to 53 months). RESULTS: Iatrogenic ulnar nerve injury occurred in two patients (3.3%), which showed complete clinical improvement in three months. Union was achieved in all the patients. Superficial pin tract infections were seen in four patients (6.6%); and cubitus varus (1.6%) and myositis ossificans in two patients, respectively. Deep infection or compartment syndrome were not encountered. The results were excellent in 49 (80.3%), good in seven (11.5%), fair in two (3.3%), and poor in three patients (5%). CONCLUSION: Closed reduction and percutaneous pinning with three K-wires is an efficacious and safe procedure with a low complication rate in the treatment of type III displaced supracondylar fractures of the humerus. PMID- 15805751 TI - [The effect of long- or short-arm casting on the stability of reduction and bone mineral density in conservative treatment of Colles' fractures]. AB - OBJECTIVES: We evaluated the effect of long- or short-arm casting on the stability of reduction and bone mineral density (BMD) in the forearm in patients treated conservatively for Colles' fractures (CF). METHODS: Eighty-three patients (48 females, 35 males; mean age 53 years; range 30 to 76 years) with an isolated unilateral CF underwent closed reduction followed by a randomly assigned long-arm (n=44) or short-arm (n=39) casting. Fractures were classified according to the Frykman's system. After reduction, radiographs of both forearms were taken, on which radial height and inclination, and volar tilt were measured and assessed according to the criteria by Sarmiento et al. In the first week, BMD measurements were made on the unaffected side to obtain reference values from four sites of the forearm, namely ultradistal, 1/3 proximal, middle diaphysis, and total. Following removal of the casts (mean 45.3 days; range 40 to 55 days), radiographic and BMD assessments were repeated. Osteoporosis was defined according to the criteria of the World Health Organization. RESULTS: The two casting groups were similar with respect to age, sex, Frykman's classification, involved side, and the dominant extremity. Osteoporosis was detected in 20% according to the T scores. All the sites in the fractured forearm showed density losses, but the difference was significant only in the middle diaphysis (p<0.05). No significant relationship was found between BMD losses and the cast type. Angular measurements showed significant deterioration after union; however, none was found to be related to the cast type (p>0.05). CONCLUSION: Our results show that BMD losses and deterioration in reduction following treatment of CF occur irrespective of which type of casting is used. PMID- 15805752 TI - [The use of local anesthesia in endoscopic release of the carpal tunnel]. AB - OBJECTIVES: We evaluated the early results of endoscopic carpal tunnel release with the use of local anesthesia. METHODS: Endoscopic carpal tunnel release was performed in 14 hands of 10 patients (8 females, 2 males; mean age 43 years; range 35 to 58 years) using the two-portal Chow technique following local anesthesia described by Wood and Logan. Involvement was in the right hand in five patients, in the left in three, and in both hands in two patients. Electromyography showed moderate or severe compression of the median nerve in the carpal tunnel and the Tinel's and Phalen's signs were positive in all the patients. There was no previous history of metabolic diseases such as diabetes mellitus, gout, or hypothyroidism, nor a coexisting pregnancy. The decision for surgery was given at the end of an unsuccessful conservative treatment for a month. The mean follow-up period was 16 months (range 10 to 24 months). RESULTS: Two patients (14.3%) required further anesthesia with local anesthetic agents and sedation in the incision area. No complaints of paresthesia were seen during injections. Injection-associated problems such as increased thickness of the synovial layer or decreased endoscopic view did not occur. No instances of tendon or nerve injury, hematoma, or sympathetic dystrophy were seen. One patient developed neuropraxia in the third and forth fingers postoperatively. Early postoperative pain was observed in two wrists, appearing 10 days and two months after surgery, respectively. During the follow-up period, all the patients were free of preoperative complaints and no recurrences were encountered. CONCLUSION: Local anesthesia is a rapid and reliable technique in carpal tunnel surgery, enabling hospitalization confined to the same day. PMID- 15805753 TI - [Treatment of AO type C2-C3 fractures of the distal end of the radius with external fixation]. AB - OBJECTIVES: We evaluated the results of combined treatment with closed reduction, grafting, K-wires, and external fixation in patients with fractures of the distal end of the radius. METHODS: Of 24 patients (15 males, 9 females; mean age 34 years; range 21-73), 16 patients had C2, eight patients had C3 fractures according to the AO classification. Three patients had open fractures, and four patients had signs of median nerve compression. Treatment included closed reduction, autologous grafting (16 cases) followed by fixation with 2 to 5 K wires, and an external fixator. Clinical and radiographic results were assessed according to the Gartland-Werley's and Stewart's criteria, respectively, and degenerative changes to the Knirk-Jupiter's criteria. The mean follow-up was 35 months (range 25 to 52 months). RESULTS: All of the 19 patients who were employed returned to preinjury working conditions in a mean of 3.5 months. Radial length was preserved to a great extent. The mean amount of collapse of the joint surface was 1.4 mm. The mean loss was 6.7% in radial inclination, and 17.6% in the volar tilt. According to the Stewart's criteria, the results were good, moderate, and poor in 12, nine, and three patients, respectively. Flexion-extension (75%), supination-pronation (70%), and radial-ulnar deviation (75%) were preserved to a great extent. Grip force amounted to 70% of the healthy side. According to the Gartland-Werley criteria, three patients had excellent, eight patients had good, 10 patients had moderate, and three patients had poor results. According to the Knirk-Jupiter's criteria, degenerative changes were slight in 11 patients, moderate in six patients, and severe in one patient. Six patients had no signs of degeneration. Complications included pin track infections (n=4), early transient reflex sympathetic dystrophy (n=2), and hypoesthesia along the superficial branch of the radial nerve (n=2). CONCLUSION: Treatment of the distal end fractures of the radius (AO type C2-C3) with a combination of external fixation, grafting, and percutaneous K-wires provides almost normal radiologic and clinical parameters. PMID- 15805754 TI - [Management of fibular hemimelia with the Ilizarov circular external fixator]. AB - OBJECTIVES: We evaluated the results of treatment with the Ilizarov circular external fixator for limb length inequality and deformities in patients with type IA, IB, and type II fibular hemimelia. METHODS: Tibial corticotomy and distraction osteogenesis with the Ilizarov technique were performed in five male patients (mean age 11.4 years; range 4 to 20 years). According to the classification of Achterman and Kalamchi, fibular hemimelia was type IA, IB, and II in three patients, one patient, and one patient, respectively. Involvement was on the right in three patients, and on the left in two patients. Two patients had equinus and one patient had valgus deformities. No instability existed in the ankle and knee joints. The mean leg discrepancy was 8.7 cm (range 3 to 16.5 cm), and the mean lengthening index was 1.6 cm/month (range 1.4 to 2 cm). The mean follow-up was 33 months (range 15 to 68 months). RESULTS: On final examinations, full range of motion of the knee was obtained. A plantigrade foot was achieved in three feet, while two sustained an equinus deformity of 17 and 15 degrees, respectively. Pin tract infections were observed in four patients, all of which were treated with oral antibiotics and dressing. During distraction, three patients had pain. Two patients had a limited range of motion of the ankle joint, without instability or subluxation of the ankle and knee joints. These joint problems were successfully dealt with by physical exercises. CONCLUSION: The Ilizarov technique is a convenient method in the correction of angular and rotational deformities while enabling distraction in type I and type II fibular hemimelia. PMID- 15805755 TI - [Our clinical experience in the treatment of snakebites]. AB - OBJECTIVES: We evaluated the results of medical and surgical treatment for venomous snakebites and reviewed current principles of first aid and therapy for affected patients. METHODS: Fourteen venomous snakebite victims (8 males, 6 females; mean age 22 years; range 7 to 75 years) were enrolled in the study. Six patients received medical treatment alone, while eight patients required both medical and surgical treatments. Injury was in the upper and lower extremities in nine and five patients, respectively. Fasciotomy was performed in seven patients due to ensuing compartment syndrome, which was manifest with extreme swelling in the affected extremity and severe pain on passive stretching of the muscles at the site of the lesion. Fasciotomy site was primarily closed in three patients, whereas four patients required debridement and skin grafting. One patient, who developed necrosis due to an excessively tight tourniquet at the time of first aid, underwent amputation of the third finger at the level of the middle phalanx. The mean follow-up was 11.5 months (range 3 to 30 months). RESULTS: Following fasciotomy, a long incision line remained in all the patients and a marked scar tissue due to skin grafting, which were associated with flexion contracture deformities in two elbows (35 and 105 degrees). Hemopericardium detected in one patient was dealt with by medical treatment. The mean length of hospital stay was 11.3 days for medically treated patients, and 18.2 days following surgical treatment. No incidence of late serum disease or mortality was encountered. CONCLUSION: Management of snakebite victims include an appropriate first aid and treatment at the hospital; identification of compartment syndrome through clinical means and measurements should lead to an indication for fasciotomy. PMID- 15805756 TI - [Psychiatric symptoms in patients treated with an Ilizarov external fixator]. AB - OBJECTIVES: The Ilizarov technique limits daily activities of patients and may cause psychiatric symptoms. We investigated psychiatric symptoms associated with the use of an Ilizarov circular external fixator (CEF) and the relationship between symptoms and duration of CEF application in adult patients. METHODS: The study included 40 adult male outpatients (mean age 26.4 years; range 20 to 40 years) who were treated with a CEF. The participants were administered a questionnaire on sociodemographic and clinical characteristics. Psychiatric symptoms were assessed using the Symptom Check List (SCL-90-R). Data were compared with those of a control group consisting of 30 healthy males with similar sociodemographic features. RESULTS: Significant differences were found between the patient and control groups with regard to somatization (p=0.03), interpersonal sensitivity (p=0.027), depression (p=0.003), anxiety (p=0.025), hostility (p=0.004), paranoid ideation (p=0.021), additional scale (p=0.000), and overall symptom (p=0.024) scores. There was at least one psychiatric symptom in 52.5% of the subjects treated with a CEF. Hostility, interpersonal sensitivity, sleep disturbances, obsessive-compulsive signs, and anxiety were detected in 37.5%, 35%, 32.5%, 20%, and 20%, respectively. The duration of CEF application was positively correlated with depression and interpersonal sensitivity (p=0.000), and negatively correlated with anxiety (p=0.001). CONCLUSION: Our data demonstrate that, during treatment with a CEF, psychiatric problems emerge, which warrant careful evaluation of the psychiatric condition of the patients. PMID- 15805757 TI - [Histopathologic and morphometric changes in rat nerve and blood vessels associated with femoral lengthening]. AB - OBJECTIVES: We investigated histopathologic and morphometric changes in rat femoral arteries, veins, and nerves associated with femoral lengthening. METHODS: The study included 42 male Wistar-albino rats. All the rats underwent left femoral osteotomy which was stabilized with an external fixator device. The rats were divided into five groups except for seven control rats which were left untreated. Femoral lengthening was performed with a distraction of 0.35 mm four times a day, which was continued for three days in group 1 (10%), and seven days in the other groups (30%). Before sacrification of each group of rats under general anesthesia after 3, 7, 14, 21, and 31 days of osteotomy, respectively, biopsy samples were obtained from distraction sites involving femoral artery, vein, and nerves for histopathologic and histomorphometric studies. RESULTS: Arterial changes observed in the first four groups were flattening and/or focal absence of the lamina elastica interna and externa, and hydropic degeneration and cytoplasmic vacuolization of the smooth muscle cells of the tunica media. Group 5 exhibited a normal arterial appearance except for fibrosis. Femoral vein changes were characterized by smooth muscle cell degeneration and cytoplasmic vacuolization in the first three groups, and by irregularities in elastic fibres and fibrosis in the remaining two. In group 5, the diameter of the femoral vein decreased by 27.9%. No histologic changes were noted in nerve morphology. In group 2, perineurium thickness and the diameter of myelinated nerve fibers notably increased by 164.6% and 58.4%, respectively. CONCLUSION: Our results show that the effect of femoral lengthening is heavily on blood vessels rather than on nerves. PMID- 15805759 TI - [Total dislodgement of the femoral component following cemented total knee arthroplasty: a case report]. AB - Total dislodgement of the components, which is the most severe form of loosening, has hitherto been unreported following total knee arthroplasty. An eighty-four year-old woman presented with complaints of pain and sensation of insecurity of her right knee after cemented total knee arthroplasty. On physical examination, a clunk was elicited during movements of the knee; however, radiographs appeared normal except for a separated fragment of the medial femoral condyle. Further examination with varus and valgus stress tests under fluoroscopic control showed dislodgement of the femoral component, which was only apparent on stress radiographs. She underwent revision cemented total knee arthroplasty and her complaints disappeared completely within a follow-up period of three years. This case illustrates the need for stress radiographs when standard radiographs are normal in the face of a high suspicion of loosening. PMID- 15805758 TI - [The factors affecting thermal necrosis secondary to the application of the Ilizarov transosseous wire]. AB - OBJECTIVES: We investigated thermal changes associated with the application of the Ilizarov transosseous wires, the extent of necrosis, and the factors affecting necrosis. METHODS: We used a pair of tibiae from a 1-year-old cow. After removal of metaphyseal areas, each of four equal diaphyseal zones marked on both tibiae was drilled at 600, 900, 1,200 and 1,800 rpm, each time with a new wire. Heat changes were recorded with heat electrodes during the application and the speed of the wire was calculated. For histopathological examination, specimens were obtained at the access and exit sites to assess the extent of necrosis. Thermal changes between the zones and immediate and remote cortices were compared. The most significant factor affecting the heat changes was analyzed by linear regression. RESULTS: Heat changes varied between 48.4 degrees C (at 1,200 rpm) and 151.9 degrees C (at 600 rpm). The thickness of the immediate cortex, the time and speed for the wire to pass the cortex were found as significant parameters in heat changes (p=0.003, p=0.01, and p=0.01, respectively). A negative correlation was found between the speed of the wire and the thickness of the necrotic area (r=-0.901, p=0.001). Regression analysis showed that the time for the wire to pass through the cortex was the most significant factor in inducing heat changes in both cortices (p=0.001, p=0.003, respectively). Histopathologically, the extent of necrosis and bone erosion was associated with lower drill speeds. Necrosis was significantly notable in the immediate cortex than that of the remote one (p=0.006). CONCLUSION: Transosseous wires should be passed at high drill speeds and with earliest time elapses to reduce thermal necrosis. PMID- 15805760 TI - [A case of lumbar ganglion cyst causing radiculopathy]. AB - Ganglion cysts represent a rare pathology mostly encountered in the lumbar region of the spinal column. Magnetic resonance imaging revealed a ganglion cyst at the L4-5 level in a 46-year-old woman who had a complaint of long-standing pain in her right leg. The cyst was completely excised following total laminectomy at L4. After surgery, her symptoms and neurological signs completely disappeared. PMID- 15805761 TI - [An isolated sacral fracture and a fracture dislocation in two pediatric patients]. AB - Isolated sacral fractures are very rare in children. Two children, aged eight and 12 years, presented with an isolated sacral type III fracture and a fracture dislocation, respectively, both of which were not associated with any neurologic problems. Without any attempts for reduction, the fractures healed completely with conservative treatment. These cases demonstrated a high remodeling potential in children for spontaneous healing. PMID- 15805762 TI - On the elusive quest for medical appropriateness. PMID- 15805763 TI - An evidence-based approach to the use of drug-eluting stents in clinical practice. AB - Less than a year after their release, drug-eluting stents are being used in more than 70% of patients who undergo percutaneous intervention for obstructive coronary disease in the United States. This unprecedented quick adoption was fueled by results of several randomized trials that demonstrated a profound reduction in restenosis rates compared with bare-metal stents. Subset analysis of the drug-eluting stent trials shows a strong restenosis reduction rate across a wide range of patient characteristics; however, these broad beneficial effects are based on randomized subjects who may not represent the actual population currently being treated with coronary stents. This review presents an analysis of the available data on the approved drug-eluting stents, including patient subsets that may or may not benefit; potential stent-specific complications; and a discussion of costs, risks, and cost-effectiveness. PMID- 15805764 TI - Advanced heart failure: a practical management algorithm and therapeutic options. AB - Despite medical and surgical advances in the management of heart failure, 3%-10% of patients clinically reside in the severe stages of this condition. The management of advanced, end-stage heart failure is one of the major challenges of medicine today. An algorithm applicable to the management of severe heart failure is presented and discussed, with an emphasis on medical treatment and therapeutic options. PMID- 15805765 TI - Selecting the best heart valve for your patient: mechanical or tissue. AB - This review provides general guidance for heart valve selection. Mechanical heart valves exhibit excellent durability and hemodynamic performance but require anticoagulation to reduce thromboembolism, and therefore risk of anticoagulation related hemorrhage is increased. Tissue valves were introduced to avoid anticoagulation, but in fact often do not, and lack durability. A literature review was performed to compare the complications of thromboembolism, anticoagulation-related hemorrhage, reoperation structural valve deterioration, and reoperative mortality associated with mechanical and tissue valves. The thromboembolism rates for mechanical and tissue valves are equivalent. During their lives, many recipients of tissue valves receive anticoagulation therapy due to comorbid conditions. The anticoagulation-related blood loss rates associated with mitral mechanical valves and mitral tissue valves are equivalent, whereas the blood loss rates associated with aortic tissue valves are less than those associated with aortic mechanical valves. PMID- 15805766 TI - Transitions: noninvasive coronary angiography using electron beam computed tomography: technique, clinical application, future prospective. AB - Electron beam computed tomography has been available clinically for 20 years. It is the only computed tomography scanner specifically developed for cardiac imaging. Over the past decade, with improvements in methodology and computer software, electron beam computed tomography has been shown to provide an excellent method to perform noninvasive coronary angiography. This article looks at the historical aspects of electron beam computed tomography and comments on how to perform and interpret electron beam angiography studies. The expanding development of noninvasive coronary and peripheral angiography methods using computed tomography will have a significant influence on cardiovascular specialists and their practices. PMID- 15805767 TI - Using the market to regulate health care price: why heart hospitals will have a competitive advantage in the world of post-diagnostic related group pricing. AB - For the past 20 years, the federal government has reimbursed hospital services by administrating pricing. Simply put, under such a system the government dictated the prices of medical services. Not only has administrative pricing failed to control medical inflation, but such failure could have been predicted from a review of basic economics. Accordingly, to eliminate the deleterious effects of administrative pricing, it is not surprising that the government is gathering information on hospital quality and cost in anticipation of a return to a system in which the price for hospital services is determined by the market. For some hospitals, this will be good news because they will be able to negotiate a more favorable rate of reimbursement. Unfortunately, for some hospitals a market system will be bad news because the government is not going to negotiate a provider contract with every hospital. In short, when the government returns to a market system for pricing of hospital services, competition among hospitals is going to become even more competitive. PMID- 15805768 TI - Transparency is a prerequisite of pay for performance. PMID- 15805769 TI - Military heart hospital at Hampstead, England: World War I. PMID- 15805770 TI - Absent septal q wave and other abnormalities. PMID- 15805771 TI - Spectral Doppler tracings--noninvasive hemodynamics. PMID- 15805772 TI - Evaluating dyspnea with myocardial perfusion imaging. PMID- 15805775 TI - As antisense RNA gets intronic. AB - Recent work describing the transcriptional output of the human genome points to the existence of a significant number of non-coding RNA transcripts coming from intronic regions, with a fraction of these being oriented antisense relative to the protein-coding mRNA of the known gene. In this article, we survey the main findings of the large-scale expression analysis projects that led to the identification of antisense intronic messages and which demonstrate their ubiquitous expression in the human genome. We review the current knowledge on long, unspliced, intronic antisense transcripts, a new class of non-coding RNAs, recently described by our group to be correlated with the degree of tumor differentiation in prostate cancer, which we postulate is involved in the fine tuning of gene expression in eukaryotes. Possible mechanisms of antisense intronic transcript biogenesis and function in gene expression regulation are discussed, as is their involvement in human diseases. While there is still no conclusive evidence demonstrating a functional role for these long, intronic antisense messages, the far-reaching implications of their existence for the mechanisms regulating gene expression certainly warrant further experimentation. PMID- 15805776 TI - Identification of putative sulfurtransferase genes in the extremophilic Acidithiobacillus ferrooxidans ATCC 23270 genome: structural and functional characterization of the proteins. AB - Eight nucleotide sequences containing a single rhodanese domain were found in the Acidithiobacillus ferrooxidans ATCC 23270 genome: p11, p14, p14.3, p15, p16, p16.2, p21, and p28. Amino acids sequence comparisons allowed us to identify the potentially catalytic Cys residues and other highly conserved rhodanese family features in all eight proteins. The genomic contexts of some of the rhodanese like genes and the determination of their expression at the mRNA level by using macroarrays suggested their implication in sulfur oxidation and metabolism, formation of Fe-S clusters or detoxification mechanisms. Several of the putative rhodanese genes were successfully isolated, cloned and overexpressed in E. coli and their thiosulfate:cyanide sulfurtransferase (TST) and 3 mercaptopyruvate/cyanide sulfurtransferase (MST) activities were determined. Based on their sulfurtransferase activities and on structural comparisons of catalytic sites and electrostatic potentials between homology- modeled A. ferrooxidans rhodaneses and the reported crystal structures of E. coli GlpE (TST) and SseA (MST) proteins, two of the rhodanese-like proteins (P15 and P16.2) could clearly be defined as TSTs, and P14 and P16 could possibly correspond to MSTs. Nevertheless, several of the eight A. ferrooxidans rhodanese-like proteins may have some different functional activities yet to be discovered. PMID- 15805777 TI - Cyanobacteria from Brazilian building walls are distant relatives of aquatic genera. AB - The 16S-rDNA from 22 cyanobacteria isolated from biofilms on walls of modern and historic buildings in Brazil was partially sequenced (approximately 350 bp) using specific primers. The cyanobacteria with the closest matching sequences were found using the BLAST tool. The sequences were combined with 52 other cyanobacterial sequences already deposited in public data banks and a dendrogram constructed, after deletion from each sequence of one of the variable 16S rDNA regions (VI). The newly sequenced organisms fitted well within their respective families, but their similarities to other members of the groups were generally low, less than 96%. Close matches were found only with one other terrestrial (hot dry desert) cyanobacterium, Microcoleus sociatus, and with Anabaena variabilis. Phylogenetic analysis suggested that the deletion of the hypervariable regions in the RNA structure is essential for meaningful evolutionary studies. The results support the standard phylogenetic tree based on morphology, but suggest that these terrestrial cyanobacteria are distant relatives of their equivalent aquatic genera and are, indeed, a distinct population. PMID- 15805778 TI - Comparative analyses of Xanthomonas and Xylella complete genomes. AB - Computational analyses of four bacterial genomes of the Xanthomonadaceae family reveal new unique genes that may be involved in adaptation, pathogenicity, and host specificity. The Xanthomonas genus presents 3636 unique genes distributed in 1470 families, while Xylella genus presents 1026 unique genes distributed in 375 families. Among Xanthomonas-specific genes, we highlight a large number of cell wall degrading enzymes, proteases, and iron receptors, a set of energy metabolism genes, second copy of the type II secretion system, type III secretion system, flagella and chemotactic machinery, and the xanthomonadin synthesis gene cluster. Important genes unique to the Xylella genus are an additional copy of a type IV pili gene cluster and the complete machinery of colicin V synthesis and secretion. Intersections of gene sets from both genera reveal a cluster of genes homologous to Salmonella's SPI-7 island in Xanthomonas axonopodis pv citri and Xylella fastidiosa 9a5c, which might be involved in host specificity. Each genome also presents important unique genes, such as an HMS cluster, the kdgT gene, and O-antigen in Xanthomonas axonopodis pv citri; a number of avrBS genes and a distinct O-antigen in Xanthomonas campestris pv campestris, a type I restriction modification system and a nickase gene in Xylella fastidiosa 9a5c, and a type II restriction-modification system and two genes related to peptidoglycan biosynthesis in Xylella fastidiosa temecula 1. All these differences imply a considerable number of gene gains and losses during the divergence of the four lineages, and are associated with structural genome modifications that may have a direct relation with the mode of transmission, adaptation to specific environments and pathogenicity of each organism. PMID- 15805779 TI - Whole-genome expression profiling of Xylella fastidiosa in response to growth on glucose. AB - Xylella fastidiosa is the etiologic agent of diseases in a wide range of economically important crops including citrus variegated chlorosis, a major threat to the Brazilian citrus industry. The genomes of several strains of this phytopathogen have been completely sequenced enabling large-scale functional studies. In this work we used whole-genome DNA microarrays to investigate the transcription profile of X. fastidiosa grown in defined media with different glucose concentrations. Our analysis revealed that while transcripts related to fastidian gum production were unaffected, colicin-V-like and fimbria precursors were induced in high glucose medium. Based on these results, we suggest a model for colicin-defense mechanism in X. fastidiosa. PMID- 15805780 TI - Large-scale, multi-genome analysis of alternate open reading frames in bacteria and archaea. AB - Analysis of over 300,000 annotated genes in 105 bacterial and archaeal genomes reveals an unexpectedly high frequency of large (>300 nucleotides) alternate open reading frames (ORFs). Especially notable is the very high frequency of alternate ORFs in frames +3 and -1 (where the annotated gene is defined as frame +1). The occurrence of alternate ORFs is correlated with genomic G+C content and is strongly influenced by synonymous codon usage bias. The frequency of alternate ORFs in frame -1 is also influenced by the occurrence of codons encoding leucine and serine in frame +1. Although some alternate ORFs have been shown to encode proteins, many others are probably not expressed because they lack appropriate signals for transcription and translation. These latter can be mis-annotated by automatic gene finding programs leading to errors in public databases. Especially prone to mis-annotation is frame -1, because it exhibits a potential codon usage and theoretical capacity to encode proteins with an amino acid composition most similar to real genes. Some alternate ORFs are conserved across bacterial or archaeal species, and can give rise to misannotated "conserved hypothetical" genes, while others are unique to a genome and are misidentified as "hypothetical orphan" genes, contributing significantly to the orphan gene paradox. PMID- 15805781 TI - In silico identification of regulatory elements of GRIN1 genes. AB - The ionotropic receptor of glutamate activated by N-methyl-D-aspartate (iGluR NMDA) is a multiheteromeric complex constituted by at least three different types of subunits, encoded by seven different genes. The subunits of iGluR-NMDA have a complex system of regulation of their gene expression. Their expression is specific for each type of neural cell, as well as for the age of the organism. Moreover, there are reports that iGluR-NMDA expression is species-specific. Even though this macromolecular complex is very important in physiology and pathology of the central nervous system, knowledge to date about the regulatory elements controlling expression is scarce. We present the results of an in silico prediction of potential regulatory elements, some of which coincide with the few known experimentally. We also present the important differences regarding the presence and the localization of the regulatory elements among human, rat, and mouse species. PMID- 15805782 TI - Exploring the genome of Trypanosoma vivax through GSS and in silico comparative analysis. AB - A survey of the Trypanosoma vivax genome was carried out by the genome sequence survey (GSS) approach resulting in 1,086 genomic sequences. A total of 455 high quality GSS sequences were generated, consisting of 331 non-redundant sequences distributed in 264 singlets and 67 clusters in a total of 135.5 Kb of the T. vivax genome. The estimation of the overall G+C content, and the prediction of the presence of ORFs and putative genes were carried out using the Glimmer and Jemboss packages. Analysis of the obtained sequences was carried out by BLAST programs against 12 different databases and also using the Conserved Domain Database, InterProScan, and tRNAscan-SE. Along with the existing 23 T. vivax entries in the GenBank, the 32 putative genes predicted and the 331 non-redundant GSS sequences reported herein represent new potential markers for the development of PCRbased assays for specific diagnosis and typing of Trypanosoma vivax. PMID- 15805785 TI - The evaluation of abnormal uterine bleeding. PMID- 15805786 TI - Pathologic diagnosis of the abnormally bleeding patient. PMID- 15805787 TI - Abnormalities of hemostasis and abnormal uterine bleeding. PMID- 15805788 TI - Endocrine mechanisms and management for abnormal bleeding due to perimenopausal changes. PMID- 15805789 TI - Role, epidemiology, and natural history of benign uterine mass lesions. PMID- 15805790 TI - Minimally invasive surgery for ablation of the endometrium. PMID- 15805791 TI - Hormonal management of abnormal uterine bleeding. PMID- 15805792 TI - Minimally invasive surgery for mass lesions. PMID- 15805793 TI - Uterine fibroid embolization. PMID- 15805796 TI - Circulating angiogenic factors in preeclampsia. PMID- 15805797 TI - Novel techniques for cerebral evaluation in preeclampsia and eclampsia. PMID- 15805798 TI - Thrombophilia and preeclampsia: the evidence so far. PMID- 15805799 TI - Prevention of preeclampsia with antioxidants: evidence from randomized trials. PMID- 15805800 TI - Expectant management of severe preeclampsia: proper candidates and pregnancy outcome. PMID- 15805801 TI - Antihypertensive medications in management of gestational hypertension preeclampsia. PMID- 15805802 TI - Controversies with the diagnosis and management of HELLP syndrome. PMID- 15805803 TI - Magnesium sulfate prophylaxis in preeclampsia: evidence from randomized trials. PMID- 15805804 TI - Long-term outcomes after preeclampsia. PMID- 15805807 TI - Which EEG patterns warrant treatment in the critically ill? Reviewing the evidence for treatment of periodic epileptiform discharges and related patterns. AB - Continuous electroencephalographic monitoring in critically ill patients has improved detection of nonconvulsive seizures and periodic discharges, but when and how aggressively to treat these electrographic patterns is unclear. A review of the literature was conducted to understand the nature of periodic discharges and the strength of the data on which management recommendations have been based. Periodic discharges are seen from a wide variety of etiologies, and the discharges themselves are electrographically heterogeneous. This spectrum suggests a need to consider these phenomena along a continuum between interictal and ictal, but more important clinically is the need to consider the likelihood of neuronal injury from each type of discharge in a given clinical setting. Recommendations for treatment are given, and a modification to current criteria for the diagnosis of nonconvulsive seizures is suggested. PMID- 15805808 TI - Continuous EEG monitoring in patients with subarachnoid hemorrhage. AB - Patients with subarachnoid hemorrhage (SAH) are at risk for seizures and delayed cerebral ischemia, both of which can be detected with continuous EEG monitoring (cEEG). Ischemia can be detected with EEG at a reversible stage. CEEG may be most useful in patients with poor grade SAH, as the neurological exam is of limited utility in these stuporous or comatose patients. Seizures have been detected in 19% of SAH patients undergoing cEEG, with the vast majority (95%) of these seizures being nonconvulsive and without any detectable clinical correlate. Applying quantitative analysis to the cEEG (relative alpha variability, post stimulation alpha/delta ratio) allows reliable detection of ischemia from vasospasm, with EEG changes often preceding changes in the clinical exam and other non-continuous monitoring techniques by up to two days. In patients at risk for developing vasospasm, cEEG monitoring, preferably with quantitative EEG analysis, should be started as early as possible and carried out for up to 14 days after the SAH. CEEG findings may lead to therapeutic (e.g., antiepileptic medication, hypertensive therapy, angioplasty) or additional diagnostic interventions such as angiography, CT or MRI. PMID- 15805809 TI - Continuous EEG monitoring for the detection of seizures in traumatic brain injury, infarction, and intracerebral hemorrhage: "to detect and protect". AB - Brain injury results in a primary pathophysiologic response that enables the brain to have seizures. Seizures occur frequently after traumatic and nontraumatic intracerebral bleeding. These seizures can be nonconvulsive, and if one does not monitor for seizures, one will not know they are occurring. The use of continuous EEG monitoring (cEEG) to detect brain arrhythmias after a primary insult, much in way that cardiac arrhythmias are detected after myocardial infarction, can influence treatment decisions and mitigate some of the pathophysiologic natural history of brain injuries. Seizures after brain injury worsen clinical outcome and need to be treated. In summary, cEEG is a valuable clinical instrument "to detect and protect," i.e., to detect seizures and protect the brain from seizure-related injury in critically ill patients, whose brains are often in a particularly vulnerable state. PMID- 15805810 TI - Continuous EEG monitoring in the intensive care unit: technical and staffing considerations. AB - Continuous EEG monitoring in the intensive care unit (ICU) is superficially similar to that which occurs in the epilepsy monitoring unit, but it also presents unique technical challenges. ICU monitoring imposes an expectation of reliability on EEG recording equipment exceeding that which may be demanded by other settings; performance requirements may also differ. Reliable network connectivity between ICUs and other hospital locations is necessary, as is provision for off-site EEG review and display. Staffing should be adequate to support continuous recording, including performing hookups at off-hours. Selection of electrode number, type, and application method requires weighting of monitoring needs, nursing concerns, and imaging requirements. PMID- 15805811 TI - New chronic EEG electrode for critical/intensive care unit monitoring. AB - The main limiting factor of EEG monitoring in the critical/intensive care environments is, and always has been, the recording electrode. The electrode and its application to the scalp has changed very little since EEG was first discovered and developed as a clinical tool. However, the evolution of amplifiers and data acquisition systems have made tremendous strides. Modern-day EEG recording systems now have the capability to record for days and weeks with little intervention, whereas the EEG electrode requires constant attention and skilled adjustment every 10 to 24 hours. If one surveys the vast array of electrodes used now and in the past, the only electrode that, once placed, never needed any further adjustment for days and weeks on end, was the chronic silver silver/chloride (Ag-Ag/Cl) sphenoidal (Sp) electrode. This Sp electrode has now been modified to permit it to be placed subdermally, similar to that of a subdermal needle electrode, but now the needle is removed to leave in place a fine, flexible, durable, chronic Ag-Ag/Cl electrode. Once placed, this subdermal wire electrode (SWE, patent pending) starts to record immediately with a low impedance of 3 to 4 Komega. This electrode can record any biopotential, in humans and in animals, and in most recording environments; it never needs adjustment, and records high-quality biopotential signals for as long as it is left in place. The SWE is also MRI and computed-tomography compatible. It takes less than half the time to place the SWE, and placement can now be performed by any medically trained personnel to obtain a low-maintenance, high-quality EEG recording. PMID- 15805812 TI - Multimodal brain monitoring in the neurological intensive care unit: where does continuous EEG fit in? AB - Continuous EEG (cEEG) is a vital component of patient monitoring in the neurologic intensive care unit, allowing the intensivist to diagnose nonconvulsive seizure activity. Though still in its infancy, Fourier-transformed cEEG data are also increasingly being used in ICUs to monitor global cerebral activity and cortical function. In conjunction with other components of multimodality neurologic monitoring, including intracranial pressure, cerebral blood flow, brain tissue oxygen tension monitoring, transcranial Doppler, and microdialysis monitoring, cEEG provides unique data regarding the electrical activity of the brain. The main challenge for clinicians and researchers will be to understand how these different aspects of multimodality monitoring relate to each other, and how physiologic variables such as blood pressure, osmolality, and temperature can be manipulated to optimize cerebral function and tissue survival in the setting of acute injury. PMID- 15805813 TI - The ACNS subcommittee on research terminology for continuous EEG monitoring: proposed standardized terminology for rhythmic and periodic EEG patterns encountered in critically ill patients. PMID- 15805814 TI - Excitability of spinal inhibitory circuits in patients with spasticity. AB - The excitability of Ia inhibition and D1 inhibition after stimulation of the common peroneal nerve to the soleus motoneuron pool was investigated in 37 spastic patients at rest and onset of voluntary ankle dorsiflexion. Ia inhibition was determined as the short-latency depression of the soleus H-reflex and D1 inhibition as the long-latency depression. There was no significant difference in Ia inhibition between the paraplegic and control groups, however Ia inhibition in the hemiplegic group was significantly decreased. D1 inhibition was reduced in the paraplegic and hemiplegic groups compared with controls. Although inhibition of the soleus H-reflex appeared at the onset of voluntary dorsiflexion in control subjects, it was not observed in the patients. Although the excitability of the Ia inhibitory pathway at rest in the patients did not differ from that in control subjects, facilitation of the Ia inhibitory pathway at the onset of movement was decreased in the patients. Ia inhibition and D1 inhibition were evaluated in two paraplegic patients who were treated with local anesthesia and surgery, respectively. The excitability of both inhibitory pathways at rest was unchanged despite improvement of reciprocal movement in one patient, and was enhanced despite reduction in muscle strength in the other patient. The excitability of spinal inhibitory pathways at rest was not always reflected by motor function in spastic patients. PMID- 15805815 TI - Intraoperative monitoring of the recurrent laryngeal nerve using acoustic, free run, and evoked electromyography. AB - The purpose of this study was to explore advantages and disadvantages of electrodes used for monitoring of the recurrent laryngeal nerve (RLN) and to determine the postoperative outcome in 135 patients. A pilot study examined 11 patients to compare the clinical performance of two commercially available recording electrodes and three stimulation electrodes for RLN neuromonitoring. After determining the most reliable and consistent stimulating/recording electrode combination, 124 patients were then monitored. In a total of 135 patients there was no permanent iatrogenic nerve damage to the RLN, although 2 patients developed transient vocal hoarseness that resolved shortly postoperatively. An initial "searching" current for evoked electromyography (EMG) was delivered at 0.7 mA and then decreased to a value of 0.35 mA, which reliably elicited RLN stimulation with a minimal incidence of false-positive results. The combination of auditory feedback from the mechanically elicited EMG and a control channel involving EMG monitoring of an additional peripheral muscle improved the detection of artifact and improved the clinical efficacy of intraoperative electromyography. A video camera showed the surgical manipulation on the EMG screen, allowing the neurophysiologist to correlate dissection with RLN status and location. Auditory responses from the nerve helped to determine the type of distress the RLN was encountering. PMID- 15805816 TI - Differences in MEG/EEG epileptic spike yields explained by regional differences in signal-to-noise ratios. AB - Controversy remains regarding the preferred modality, magnetoencephalography (MEG) or EEG, for the presurgical evaluation of patients with epilepsy. In general, it appears that the spike yields for MEG and EEG are similar in patients with temporal lobe epilepsy, and that for neocortical epilepsy the MEG spike yields may be larger than for EEG. In general, MEG/EEG spike yields depend on factors such as (1) the number of sensors, (2) the source depth and orientation, (3) the background activity, and (4) the smearing of the potential fields due to variations in skull resistivity in EEG. Because the contribution of all these factors are of the same order of magnitude, the authors took them all into account to predict the signal-to-noise ratio (SNR) of hypothetical spikes in different brain areas. In this study, it was assumed that spike sensitivity (and therefore the spike yield) increases with SNR. The estimated SNR values at temporal areas were comparable for MEG and EEG, which is in agreement with clinical findings that spike yields in temporal lobe epilepsy are similar. Furthermore, the SNR of MEG was substantially higher in the frontal area, indicating that in frontal lobe epilepsy MEG may be highly relevant to prescreening of epilepsy patients. This model-based approach indicates that SNR mapping clarifies differences between MEG and EEG findings that are difficult to understand on the basis of patient studies only. PMID- 15805817 TI - Is anyone really M.A.D.?: the occurrence and course of mixed anxiety-depressive disorder in a sample of primary care patients. AB - We examined the occurrence and 1-year course of mixed anxiety-depressive disorder (MAD) in a sample of primary care patients. Participants are part of the Primary Care Anxiety Project, a naturalistic, longitudinal study of anxiety disorders in primary care. Participants completed a questionnaire screening for anxiety symptoms. Those screening positive were invited for an interview to diagnose MAD and DSM-IV Axis I disorders. Participants were then interviewed at 6 and 12 months postintake. Of 1634 participants completing an intake interview, four participants (0.2%) met complete DSM-IV MAD criteria. The adjusted probability of remitting from MAD in 1 year was 80%. Although this was not a prevalence study, results indicate a very low occurrence of MAD across 15 primary care settings. Further, they indicate that this diagnosis may not be stable across time and raise doubts about its utility. PMID- 15805818 TI - The impact of personality on symptom expression in obsessive-compulsive disorder. AB - Previous research conducted on the five-factor model of personality (FFM) in obsessive-compulsive disorder (OCD) has demonstrated that community and clinical participants score significantly higher than controls on the domains and facets of neuroticism and extraversion and selective facets of agreeableness and conscientiousness. However, studies have yet to examine the extent to which personality traits, as assessed by the FFM, are associated with the specific symptoms of OCD. The purpose of this study was to examine further the personality predictors of obsessive-compulsive symptoms in clinical participants using the facets of the FFM. Patients with a DSM-IV diagnosis of OCD (N = 56) completed the Revised NEO Personality Inventory, the Yale Brown Obsession Compulsion Scale, and the Beck Depression Inventory. Lower scores on openness to ideas were uniquely associated with greater obsession severity, whereas lower openness to actions was uniquely associated with greater compulsive severity. In contrast with past research that has emphasized the association between neuroticism and extraversion and dimensionally rated obsessive-compulsive symptoms, this study demonstrates the specific associations between selective facet traits of openness and clinical obsessions and compulsions. Whereas tendencies toward negative affectivity may confer a nonspecific vulnerability to the development of OCD, facets of openness may impact on the particular expression and severity of obsessive-compulsive symptoms. PMID- 15805819 TI - Peer suicidal behavior and adolescent risk behavior. AB - The relationship between adolescent suicide attempts and death by suicide and psychosocial functioning of peers remains poorly understood, especially in the myriad ways that these suicidal behaviors might impact friends. This study explored the relationship between peer suicidal behavior and adolescent risk behavior using a large, nationally representative sample of adolescents (N = 5852). Results indicate that youth exposed to peer suicidal behavior are significantly more likely to have their own suicidal ideation and attempts, and to smoke cigarettes and marijuana, binge drink, be involved in a serious physical fight, and have inflicted injuries that require medical attention. These results highlight the need for professionals to be aware of these risks in friends of those who have attempted or died by suicide. Assessment and intervention for peers is appropriate and required for this at-risk group. PMID- 15805820 TI - Narrative qualities in schizophrenia: associations with impairments in neurocognition and negative symptoms. AB - Schizophrenia spectrum disorders often involve a profound diminishment in people's ability to coherently narrate their lives. Unknown is whether narrative difficulties of persons with schizophrenia differ from those with other disabilities, and how they relate to clinical or neurocognitive aspects of schizophrenia. To address both issues, personal narratives were obtained for 25 participants with schizophrenia spectrum disorders and a comparison group of eight legally blind participants and four with major depressive disorders. Flexibility of abstract thought and positive, negative, and emotional discomfort symptoms were assessed among the schizophrenia group. ANCOVA comparing scores on our scale to assess narrative development, controlling for age, found the schizophrenia group narratives significantly more impoverished than the comparison group on multiple dimensions, including self-worth and agency. Neurocognitive impairment and negative symptoms were significant predictors of impoverishment in the schizophrenia group. Results may highlight deficits that could be addressed in psychotherapy or rehabilitation to promote recovery. PMID- 15805821 TI - Religiosity/spirituality and pain in patients with sickle cell disease. AB - Religion/spirituality has been identified by individuals with sickle cell disease (SCD) as an important factor in coping with stress and in determining quality of life. Research has demonstrated positive associations between religiosity/spirituality and better physical and mental health outcomes. However, few studies have examined the influence religiosity/spirituality has on the experience of pain in chronically ill patients. Our aim was to examine three domains of religiosity/spirituality (church attendance, prayer/Bible study, intrinsic religiosity) and evaluate their association with measures of pain. We studied a consecutive sample of 50 SCD outpatients and found that church attendance was significantly associated with measures of pain. Attending church once or more per week was associated with the lowest scores on pain measures. These findings were maintained after controlling for age, gender, and disease severity. Prayer/Bible study and intrinsic religiosity were not significantly related to pain in our study. Positive associations are consistent with recent literature, but our results expose new aspects of the relationship for African American patients. We conclude that religious involvement likely plays a significant role in modulating the pain experience of African American patients with SCD and may be an important factor for future study in other populations of chronically ill pain sufferers. PMID- 15805822 TI - Predictors of development of adult psychopathology in female victims of childhood sexual abuse. AB - The purpose of this study is to identify predictors of resilience and adult mental disorders in women with a history of childhood sexual abuse. This cross sectional study was conducted in a family practice center using adult female (age 18-40) patients. Outcome measures assessed the prevalence of major depressive episode, panic disorder, agoraphobia, substance abuse, posttraumatic stress disorder, borderline personality disorder, bulimia, and suicidality. Seventy-six percent of the 90 women with sufficient data met criteria for at least one adult disorder. Mental health was related to high SES, lack of family alcohol abuse, lower frequency of first perpetrator abuse, and few perpetrators. Specifics of the abuse were associated with development of borderline personality disorder, substance abuse, major depressive episode, suicidality, bulimia, agoraphobia, and panic disorder. Maternal violence against the father, substance abuse within the household of origin, and maternal care and overprotection were also important. The specifics about the abuse and the family environment during childhood are important predictors of adult psychopathology. PMID- 15805823 TI - Factorial structure of the hallucinatory experience: continuity of experience in psychotic and normal individuals. AB - Examination of the distribution of the hallucinatory experience may aide in the determination of their continuity and the psychological mechanisms that mediate their occurrence. Past investigators have found that hallucinatory experiences are not limited to disordered individuals and can be induced in the laboratory and occur naturally in the general population. Few reports to date, however, have directly investigated the continuity of the experience by comparing hallucinatory behavior of psychotic patients with a nonclinical sample. In the present study, we examined the architecture of the hallucinatory experience by comparing the factorial structure of the Launay-Slade Hallucination Scale using psychotic patients with active hallucinations, psychotic inpatients without hallucinations, and a group of university students. In support of the continuum model of psychosis, a very similar factor-analytic solution was obtained for all three groups. Discriminant function analysis, however, revealed that all groups achieved a high classified rate by their item responses. These results are consistent with the notion that expression of hallucinatory behavior exists along a continuum, but at a certain level of symptom severity beyond a critical threshold, the behavior becomes discontinuous and dysfunctional. PMID- 15805824 TI - Gender differences and related factors affecting online gaming addiction among Taiwanese adolescents. AB - The aim of this study was to evaluate the extent to which gender and other factors predict the severity of online gaming addiction among Taiwanese adolescents. A total of 395 junior high school students were recruited for evaluation of their experiences playing online games. Severity of addiction, behavioral characteristics, number of stressors, and level of satisfaction with daily life were compared between males and females who had previously played online games. Multiple regression analysis was used to explore gender differences in the relationships between severity of online gaming addiction and a number of variables. This study found that subjects who had previously played online games were predominantly male. Gender differences were also found in the severity of online gaming addiction and motives for playing. Older age, lower self-esteem, and lower satisfaction with daily life were associated with more severe addiction among males, but not among females. Special strategies accounting for gender differences must be implemented to prevent adolescents with risk factors from becoming addicted to online gaming. PMID- 15805828 TI - Introduction: First International Symposium on: pediatric inflammatory bowel disease: September 16-18, 2003; Irvington, Virginia, U.S.A. PMID- 15805825 TI - Weight loss and PTSD symptom severity in former POWs. AB - To determine the relationship between weight loss suffered by former prisoners of war during captivity during World War II and the Korean Conflict and current posttraumatic stress disorder (PTSD) symptoms, the Clinician-Administered PTSD Symptom Scale, a lifetime stressor checklist, and the Structured Clinical Interview for DSM-IV were administered to 102 former prisoners of war. Preconfinement and postconfinement weights and length of confinement were obtained from military medical records. Percentage of body weight lost during captivity was significantly higher in those subjects with PTSD and correlated with current PTSD symptom severity. Length of confinement was not associated with current PTSD symptoms. PMID- 15805829 TI - Thymic functions related to the pathogenesis of IBD. PMID- 15805830 TI - Innate immunity. PMID- 15805831 TI - Receptor-mediated lymphocyte apoptosis in health and disease. PMID- 15805832 TI - Control of adaptive immunity: from naive to memory. PMID- 15805833 TI - Proceedings of the First International Symposium on Pediatric Inflammatory Bowel Disease. September 16-18, 2003. Irvington, Virginia, U.S.A. PMID- 15805834 TI - Costimulation and regulation of autoimmunity and tolerance. PMID- 15805836 TI - Early and late inflammatory responses. PMID- 15805835 TI - NF-kappa B, an inhibitor of microflora-induced colitis. PMID- 15805837 TI - Downstream effector functions of T-cell activation. PMID- 15805838 TI - Role of APRIL in the mucosal Iga antibody response. PMID- 15805839 TI - A genomic view of our symbiosis with members of the gut microbiota. PMID- 15805840 TI - Molecular investigation of the human gastrointestinal ecosystem. PMID- 15805841 TI - Role of commensal enteric bacteria in the pathogenesis of immune-mediated intestinal inflammation: lessons from animal models and implications for translational research. PMID- 15805842 TI - Early enterocyte responses to enteropathogenic E. coli. PMID- 15805843 TI - Signal transduction by bacterial proteins. PMID- 15805844 TI - Effector and regulatory CD4+ T cell function in a murine model of Helicobacter hepaticus-induced colitis. PMID- 15805845 TI - The hygiene hypothesis: epidemiologic evidence. PMID- 15805846 TI - Gene-environment interactions in the etiopathogenesis of inflammatory bowel disease. PMID- 15805847 TI - Opportunities in pediatric IBD research. PMID- 15805848 TI - Impact of prebiotics and probiotics on enteric flora. PMID- 15805849 TI - Celiac disease as a model of gastrointestinal inflammation. PMID- 15805851 TI - Genetics and pediatric diseases. PMID- 15805850 TI - TIM-1, hepatitis A virus and the hygiene theory of atopy: association of TIM-1 with atopy. PMID- 15805852 TI - Genomic signatures of immune and inflammatory responses. PMID- 15805853 TI - Defining molecular targets for drug design: synergism between academia and industry. PMID- 15805854 TI - Stem cell plasticity: tools for investigation and repair. PMID- 15805855 TI - Neonatal T cell function. PMID- 15805856 TI - Inverting the therapeutic triangle. PMID- 15805857 TI - The protective effect of infections on immune disorders. PMID- 15805858 TI - A pivotal role for PPR gamma in innate immune homeostasis? PMID- 15805859 TI - Flexible endoscopic evaluation of swallowing with sensory testing in patients with unilateral vocal fold immobility: incidence and pathophysiology of aspiration. AB - OBJECTIVES/HYPOTHESIS: The objective was to examine the incidence and pathophysiology of aspiration in patients with unilateral vocal fold immobility presenting with dysphagia. STUDY DESIGN: Retrospective review of flexible endoscopic evaluation of swallowing with sensory testing (FEESST) data and medical records in two tertiary medical care centers. METHODS: The data for all patients with unilateral vocal fold immobility who underwent FEESST between 2000 and 2003 were reviewed. RESULTS: Eighty-one patients (45 male and 36 female patients) were included in the study. The mean age was 59 years. The most common causes or origins were iatrogenic (42%), malignancy (23%), and neurological (18%). The immobility was left-sided in 59% of patients. A majority of the patients exhibited laryngeal edema/erythema (90%), difficulty with secretions (60%), and decreased laryngopharyngeal sensation (83%). The laryngeal adductor reflex was absent in 34% of the patients. An aspiration rate of 35% was detected with thin liquids. Trials of purees revealed a 76% rate of pooling, 44% rate of spillage, 32% rate of penetration, 18% rate of aspiration, and 24% rate of regurgitation. Rates of penetration and aspiration with purees were significantly higher in patients who had decreased laryngopharyngeal sensation, absent pharyngeal squeeze, and absent laryngeal adductor reflex. CONCLUSION: Dysphagia in patients with unilateral vocal fold immobility is demonstrated during FEESST by pooling, spillage, penetration, and aspiration. The pathophysiology of dysphagia is multifactorial with decreased sensation and limitation of airway protective mechanisms both acting as contributing factors. PMID- 15805860 TI - Effectiveness of chemotherapy and radiotherapy in sterilizing cervical nodal disease in squamous cell carcinoma of the head and neck. AB - OBJECTIVE: Determine effects of chemoradiotherapy on nodal disease in head and neck squamous cell carcinoma (SCCA). STUDY DESIGN: Matched case-control study. METHODS: A series of neck dissections (ND) performed for SCCA of the head and neck was retrospectively reviewed. Three groups were identified: 1) planned ND after chemoradiotherapy, 2) ND after radiotherapy alone, and 3) ND before adjuvant therapy (control group). Demographic data, tumor-node-metastasis stage, and pathology were reviewed. Total number of nodes recovered, number of positive nodes, and extracapsular spread (ECS) were recorded. To each patient in the chemoradiotherapy group, a randomly matched dissection in the control group was identified, matching for preoperative N stage and ND type (comprehensive, supraomohyoid, or selective). Comparisons were conducted for total nodes, presence of positive nodes, and ECS. Similar matched comparisons were conducted for ND after radiotherapy alone versus the control group. RESULTS: Ninety-seven NDs (N0 = 10 cases, N1 = 9, N2 = 69, and N3 = 9) were matched to control NDs without previous therapy. Total nodal yield was not statistically different between chemoradiotherapy and control groups (23.5 vs. 23.0 nodes, respectively, P = .77). Positive nodal yield was significantly lower for chemoradiotherapy versus control dissections (0.76 vs. 3.0, P < .001). The percent of chemoradiotherapy dissections recovered as positive nodes was 24.7% versus 68.0% of the control group (P < .001). ECS was identified in 13.4% of the chemoradiotherapy group versus 32.0% of the control group (P = .002). Radiotherapy alone produced less pronounced nodal reductions. CONCLUSIONS: Chemoradiotherapy substantially decreases nodal disease and ECS in SCCA. However, a significant percentage of necks contain positive nodes after therapy, meriting consideration for ND. PMID- 15805861 TI - Tympanomastoid suture and digastric muscle in cadaver and live parotidectomy. AB - OBJECTIVES/HYPOTHESIS: To prove that the tympanomastoid suture (TMS) is a significantly closer and less variable anatomic landmark to the facial nerve than the posterior-superior margin of the posterior belly of the digastric muscle (PBD) in parotid surgery. MATERIALS AND METHODS: A prospective study of 14 cadaver specimens and 22 live patients comparing the closest measured distances between the TMS and PBD to the facial nerve. RESULTS: The mean closest distances from the TMS and PBD to the facial nerve were 1.8 (range 0-4) mm and 12.4 (range 7-17) mm, respectively (P < .05) for cadavers. The mean closest distances in live patients from the TMS and PBD to the facial nerve were 2.0 (range 0-4) mm and 10.7 (range 5-14) mm, respectively (P < .05). CONCLUSIONS: Facial nerve identification and preservation is the key to successful parotid surgery. The TMS is a significantly closer and less variable anatomic landmark compared with the PBD both in cadaver dissection and in live patients. PMID- 15805862 TI - Bacterial biofilms in surgical specimens of patients with chronic rhinosinusitis. AB - OBJECTIVES: Biofilms are bacterial pathogens that organize in several chronic and recalcitrant infectious processes. We hypothesize that biofilms play a role in chronic rhinosinusitis (CRS). Our goal is to demonstrate biofilms in mucosal specimens of patients undergoing surgery for CRS. STUDY DESIGN: A prospective study of the presence of biofilms in patients undergoing endoscopic sinus surgery for CRS compared with control patients without CRS. METHODS: There were a total of 30 subjects and 4 controls enrolled. The samples of 24 subjects and 4 controls were cultured and then prepared using standard methods for scanning electron microscopy (SEM). The remaining six subjects' samples were treated using advanced cryofixation methods as preparation to preserve structure for SEM and transmission electron microscopy (TEM). RESULTS: Using strict SEM morphologic criteria, 24 (80%) of the 30 patients were found to have micrographic evidence of biofilms. All controls had healthy appearing cilia and goblet cells without biofilms. The six cryofixation samples showed biofilm structures on SEM micrographs that were correlated with bacterial structures seen at the mucosal surface on the corresponding TEM cross sections. Bacterial cultures were positive on all patients. CONCLUSIONS: Biofilms were demonstrated to be present in patients undergoing surgery for CRS; none of the patients without CRS had any evidence of biofilms. Although SEM is capable of demonstrating the biofilms' three-dimensional structure, glycocalyx, and water channels, it cannot clearly demonstrate the presence of bacteria within the biofilm. We were able to demonstrate evidence of bacteria in the biofilms on the subjects tested using TEM. PMID- 15805863 TI - Polyglycolic acid/poly-L-lactic acid copolymer use in laryngotracheal reconstruction: a rabbit model. AB - OBJECTIVE: To evaluate the tissue response and resorption of the polyglycolic acid/poly-L-lactic acid (PGA/PLLA) implant in laryngotracheal reconstruction and compare its dynamic stability with autologous cartilage grafts. STUDY DESIGN: An interventional, before-after trial. METHODS: Twenty-one white, female, New Zealand rabbits were divided into four groups. Groups A and B underwent laryngotracheoplasty using the PGA/PLLA implants of 3 and 4 mm widths. Group C received autologous ear cartilage grafts. Group D was the control group and did not undergo surgery. The subjects were sedated at 12 months, and the larynges were evaluated in vivo for stability and area measurements by way of endoscopy during spontaneous respiration. The subjects were then killed, the larynges harvested, and the negative intraluminal pressures applied to the laryngotracheal unit were measured in a closed-system apparatus. The larynges were then evaluated for inflammatory reaction and implant resorption by way of histologic analysis. RESULTS: All implanted subjects survived without complications and grew normally. There was no appreciable subglottic collapse during spontaneous respiration under anesthesia. Ex vivo examination of maximum negative intraluminal pressures (-50 cm H2O) in a closed system demonstrated subglottic collapse of 78%, 72%, 61%, and 3% for groups A, B, C, and D, respectively, revealing the inherent weakness in the surgically manipulated airways regardless of grafting material. Histologically, the PGA/PLLA implants were essentially completely resorbed. CONCLUSIONS: PGA/PLLA appears to be a safe and effective synthetic material for use in laryngotracheal reconstruction in the rabbit model while avoiding donor site morbidity and additional operative time. Reconstructed airways maintained adequate strength and patency under physiologic conditions and are comparable with autologous cartilage grafts. PMID- 15805864 TI - Posterior epistaxis: identification of common bleeding sites. AB - OBJECTIVE: The objective of this study was to determine common bleeding sites in the nasal cavity of patients with posterior epistaxis and thus review our management protocol. STUDY DESIGN: A prospective study was carried out from 1989 to November 2003 in the otolaryngology-head and neck surgery department of a tertiary referral center. This study included patients who presented with posterior epistaxis uncontrolled with standard nasal packing and with no identifiable bleeding point on examination under local anesthesia. METHOD: All patients underwent a formal examination under general anesthesia by the senior author of this article. Findings at examination were documented along with subsequent management and its outcome. RESULTS: Forty-three patients were included in this study. Bleeding points were identified in 36 cases. Seven patients had septal bleeding points (20%). The rest were located on the lateral nasal wall (81%). Of these, 4 were on the lateral wall of inferior meatus, 7 on the lateral surface of inferior turbinate, 8 on the lateral wall of middle meatus, and 10 on the lateral surface of middle turbinate. All were located posteriorly. CONCLUSIONS: We recommend examination under general anesthesia when conservative measures fail to control bleeding, concentrating on the posterior aspect of the lateral nasal wall. In addition, the lateral aspect of the middle and inferior turbinates may contain a groove within which bleeding points may be concealed. The lateral position of most bleeding sites indicates that use of nasal packing can only attempt to indirectly tamponade blood flow and is rarely justified bilaterally. Electrothermocautery can achieve excellent results with minimal complications. Failure to identify a bleeding point, after thorough examination under general anesthesia, does not require further intervention unless complicated by further bleeding. PMID- 15805865 TI - Postoperative bleeding after diathermy and dissection tonsillectomy. AB - OBJECTIVE: To analyze the incidence and pattern of bleeding after tonsillectomy performed by either cold dissection or diathermy. DESIGN: A prospective, nonrandomized cohort study of postoperative hemorrhage after tonsillectomy. METHODS: Monthly reporting of the number of tonsillectomies and postoperative bleeds from otolaryngologists working in rural areas of Victoria, Australia over a 2.5 year period. Criteria for bleeding were either 1) repeat anesthesia and surgery because of hemorrhage (including return to theater from the recovery room), or 2) readmission to hospital because of bleeding, or 3) blood transfusion to replace blood loss. Main outcome measures were the incidence, volume, and time course of postoperative hemorrhage. RESULTS: The number of bilateral tonsillectomies with removal by cold-blunt dissection was 3,087. In this group, there were 57 (1.85%) bleeds. The number of bilateral tonsillectomies with removal by diathermy dissection was 1,557. In this group, there were 37 (2.38%) bleeds. If cold dissection is taken as the "control" and diathermy tonsillectomy as the "treatment" group, the relative risk of bleeding after diathermy tonsillectomy is 1.30 (95% confidence interval 0.88-1.93). The pattern of bleeding after each technique differs significantly over time, with more reactionary bleeds in the dissection group and more bleeds between 4 to 7 postoperative days after diathermy. When bleeding occurred, it was in excess of 500 mL in 16% of dissection cases and 43% of diathermy tonsillectomies. CONCLUSIONS: The difference in the risk of bleeding after each technique did not reach statistical significance, but the temporal pattern of hemorrhage differed, and more bleeds exceeding 500 mL were seen in the diathermy group. PMID- 15805866 TI - Behavioral inhibition and clinical outcomes in children with cochlear implants. AB - OBJECTIVES/HYPOTHESIS: Individual speech and language outcomes of deaf children with cochlear implants (CIs) are quite varied. Individual differences in underlying cognitive functions may explain some of this variance. The current study investigated whether behavioral inhibition skills of deaf children were related to performance on a range of audiologic outcome measures. DESIGN: Retrospective analysis of longitudinal data collected from prelingually and profoundly deaf children who used CIs. METHODS: Behavioral inhibition skills were measured using a visual response delay task that did not require hearing. Speech and language measures were obtained from behavioral tests administered at 1-year intervals of CI use. RESULTS: Female subjects showed higher response delay scores than males. Performance increased with length of CI use. Younger children showed greater improvement in performance as a function of device use than older children. No other subject variable had a significant effect on response delay score. A series of multiple regression analyses revealed several significant relations between delay task performance and open set word recognition, vocabulary, receptive language, and expressive language scores. CONCLUSIONS: The present results suggest that CI experience affects visual information processing skills of prelingually deaf children. Furthermore, the observed pattern of relations suggests that speech and language processing skills are closely related to the development of response delay skills in prelingually deaf children with CIs. These relations may reflect underlying verbal encoding skills, subvocal rehearsal skills, and verbally mediated self-regulatory skills. Clinically, visual response delay tasks may be useful in assessing behavioral and cognitive development in deaf children after implantation. PMID- 15805867 TI - Fungal-specific humoral response in eosinophilic mucus chronic rhinosinusitis. AB - OBJECTIVES/HYPOTHESIS: An immunoglobulin (Ig)E-mediated allergic pathogenesis is presumed in allergic fungal sinusitis (AFS), yet extensive polyps and eosinophilic mucus (EM) in the paranasal sinuses may also occur in the absence of allergy. Although a noninvasive fungal pathogenesis is presumed in all chronic rhinosinusitis with EM (EMCRS), fungal-specific nonallergic immune responses have not been thoroughly investigated. We tested the hypothesis that there is a fungal specific humoral response in EMCRS and that it is not confined to IgE. STUDY DESIGN: EMCRS patients were prospectively stratified into subgroups based on the presence or absence of fungi within EM and of fungal-specific systemic IgE. There were 12 AFS, 5 AFS-like, 8 nonallergic fungal eosinophilic sinusitis (NAFES), and 5 nonallergic, nonfungal eosinophilic sinusitis (NANFES) patients. METHODS: Alternaria alternata and Aspergillus fumigatus-specific serum IgE, IgG, IgM, and IgA was measured by enzyme-linked immunosorbent assay and compared with strictly defined healthy and disease-control groups. RESULTS: Fungal-specific IgG (Alternaria alternata P = .0002; Aspergillus fumigatus P = .004), and IgA levels (Alternaria alternata P = .0016; Aspergillus fumigatus P = .002) were higher in EMCRS compared with healthy volunteers but not with disease controls. Fungal specific IgG3 levels were significantly elevated in all the EMCRS subgroups compared with controls for either fungal antigen (P < .0001). Importantly, fungal specific IgE levels were not significantly different between fungal-allergic EMCRS and disease controls. CONCLUSIONS: Fungal-specific immunity characterized by serum IgG3 and not IgE, distinguished the EMCRS subgroups from control groups regardless of the presence of fungus within EM or of systemic fungal allergy. Fungal-specific IgE responses in fungal-allergic EMCRS were no different to those in fungal-allergic controls, thus challenging the presumption of a unique pathogenic role of fungal allergy in "allergic fungal sinusitis." PMID- 15805868 TI - Usefulness of delayed postcontrast magnetic resonance imaging in the detection of residual cholesteatoma after canal wall-up tympanoplasty. AB - OBJECTIVES/HYPOTHESIS: Imaging takes an increasing place in the follow-up of patients who have undergone surgery for cholesteatoma, with computed tomography (CT) as the first line imaging technique. However, in case of complete opacity of the tympanomastoid cavities, CT is not able to differentiate residual cholesteatoma from postoperative scar tissue. The aim of this study was to assess the usefulness of magnetic resonance imaging (MRI) using delayed postcontrast T1 weighted images for the detection of residual cholesteatoma after canal wall-up tympanoplasty (CWU) in cases where CT was not conclusive. STUDY DESIGN: Prospective study. METHODS: MRI, with delayed postcontrast T1-weighted images (30 45 minutes after contrast injection), was performed before revision surgery in 41 consecutive patients who had undergone CWU for cholesteatoma and presenting with a nonspecific complete opacity of the mastoid bowl on CT. In all the cases, imaging results were compared with operative findings at surgical revision. RESULTS: A residual cholesteatoma was found in 19 of 41 patients at revision surgery and was correctly detected on MRI in 17 patients. In the two remaining cases, cholesteatoma pearls smaller than 3 mm were not seen. There was no false positive case. Statistics were as follows: sensitivity 90%; specificity 100%; positive predictive value 100%; negative predictive value 92%. CONCLUSION: When postoperative CT is not conclusive because of complete opacity of the tympanomastoid cavities, MRI with delayed postcontrast T1-weighted images is a reliable additional technique for the detection of a residual cholesteatoma when its diameter is at least 3 mm. PMID- 15805869 TI - Cricothyroid approximation and subluxation in 21 male-to-female transsexuals. AB - OBJECTIVES: To evaluate the medium-term outcome of cricothyroid approximation and subluxation (CTAS) with postoperative speech therapy for pitch elevation in male to-female transsexuals. STUDY DESIGN: Retrospective study of male-to-female transsexuals who underwent pitch-raising surgery between November 1996 and August 2001. METHODS: Twenty-one male-to-female transsexuals opted for surgical feminization of their voices after inadequate improvements with speech therapy alone. Electrolaryngographic measurements were obtained by a single speech therapist of modal fundamental frequencies and the percentage of irregularities before, at 2 weeks, and 6 months after surgery. All 21 patients underwent CTAS, and 20 underwent simultaneous cosmetic thyroid chondroplasty by a single surgeon. RESULTS: Electrolaryngographic results 2 weeks after surgery showed an average postoperative gain in modal frequency of free speech of 71.05 Hz (95% confidence interval [CI]: 42.9-99.2, P < .001). There was a concomitant average rise in irregularities of 9.9% (95% CI 0.7-18.5, P = .03). At median follow-up of 6 months after six sessions of speech therapy (n = 15), there was a decrease in irregularities to preoperative levels. The overall gain at 6 months in modal frequency of free speech was 56.9 Hz (95% CI 38.3-75.4, P < .001). Smoking and age did not predict a worse outcome. CONCLUSIONS: Cricothyroid approximation effectively raises pitch in male-to-female transsexuals. There is a concomitant rise in voice irregularities that is effectively addressed by speech therapy. PMID- 15805870 TI - Neuronal survival and glial reactions after recurrent laryngeal nerve resection in the rat. AB - OBJECTIVES/HYPOTHESIS: Complete axonal injury to the recurrent laryngeal nerve (RLN) leads to permanent loss of coordinated function of the intrinsic muscles of the larynx. The aim of the present study was to investigate retrograde reactions, neuronal survival, and glial reactions in the nucleus ambiguus after a distal resection of the RLN to evaluate the potential need for neuroprotective substances. STUDY DESIGN AND METHODS: A segment of the left RLN was resected in 31 adult rats. Before sacrifice of the animals at 2 to 28 days postlesion, the motor neurons in the nucleus ambiguus were retrogradely traced by the use of Fluorogold. Brainstems were isolated and processed for neuron quantification and immunohistochemical analysis. Neuron counts were performed in the nucleus ambiguus on serial sections. Glial reactions were investigated in the nucleus ambiguus using immunohistochemistry. RESULTS: No decrease in the number of motor neurons in the nucleus ambiguus could be demonstrated up to 1 month postlesion. Astroglia and microglia showed increased immunoreactivity at 7 to 14 days postinjury, followed by a slight decline in glial reaction. Microglia revealed no signs of transformation into macrophages during the study period, further indicating the absence of neuronal loss. CONCLUSIONS: Neuronal death does not occur within 1 month postlesion as a result of resection of the RLN in the adult rat, and neuroprotective substances should therefore be of minor value after RLN injury. Glial reactions appear in a similar fashion as after other peripheral nerve lesions not causing neuronal loss. PMID- 15805871 TI - Elective transcervical superior mediastinal lymph node dissection for advanced laryngeal and level 4 N3 squamous cell carcinoma. AB - OBJECTIVES: To review our results with elective superior mediastinal lymph node dissections in patients with advanced laryngeal squamous cell carcinoma (SCCA) and overt level 4 adenopathy. STUDY DESIGN: Retrospective review. METHODS: We searched operative case logs for all patients treated with an elective superior mediastinal dissection by the senior author (Y.D.) during a 7-year period. Charts were reviewed for demographic information, prior treatment with chemotherapy or radiotherapy, and pathologic results. RESULTS: Fifty-six patients who underwent elective superior mediastinal lymph node dissection for advanced laryngeal SCCA and overt level 4 adenopathy were reviewed, and superior mediastinal disease was present in 15 of 56 (26.8%) patients. Superior mediastinal nodes were positive in 11 of 42 (26.2%) patients with advanced laryngeal SCCA and 4 of 14 (28.6%) patients with N3 SCCA involving level 4. Patients with SCCA receiving prior chemotherapy and radiotherapy had a significantly higher rate of positive superior mediastinal nodes (10/21 patients, 47.6%) compared with patients without prior therapy (5/35 patients, 14.3%, P = .01, Fisher's exact test). There were no stomal recurrences in 42 patients treated for advanced laryngeal cancer. CONCLUSIONS: Elective transcervical superior mediastinal dissection was positive in 26.8% of patients with advanced laryngeal cancer or N3 disease in level 4. A transcervical superior mediastinal dissection may be safely performed without a sternotomy. PMID- 15805872 TI - The influence of lymph node metastasis in the treatment of squamous cell carcinoma of the oral cavity, oropharynx, larynx, and hypopharynx: N0 versus N+. AB - OBJECTIVES/HYPOTHESIS: Management of the N0 neck is a continuing controversy. The study compares the influence of N0 and N+ disease on the results of treating squamous cell carcinoma (SCCA) of the oral cavity (OC), oropharynx (OP), larynx (LX), and hypopharynx (HP) with five different treatment modalities. The study also compares the results of four different approaches to the treatment of the N0 neck. STUDY DESIGN: A retrospective study of 3887 patients. METHODS: Patients in the Tumor Research Project of the Department of Otolaryngology-Head and Neck Surgery of the Washington University School of Medicine (St. Louis, MO) with biopsy-proven previously untreated SCCA of one of the four above-mentioned regions who were treated with curative intent by one of five modalities and who were eligible for 5-year follow-up were included in the study. The treatment modalities included local resection of primary alone (LR), composite resection (primary with neck dissection) (CR), radiation therapy alone (RT), local resection with radiation therapy (LR/RT), and composite resection with radiation therapy (CR/RT). The N0 neck was treated with one of four approaches: observation with close follow-up reserving treatment only for subsequent neck disease, neck dissection, RT to the neck region, and a combination of neck dissection with RT. Multiple diagnostic, treatment, and follow-up parameters were studied using standard statistical analyses to determine statistical significance. RESULTS: The 5-year disease-specific survival (DSS) for the all-sites group (ASG) was 59%. The DSS for the subsites included the following: OC, 53%; OP, 47%; LX, 70%; and HP, 42%. Patients with N0 disease had significantly better DSS than patients with N+ disease at all sites. Occult neck disease in N0 patients was low with 4% pN1 for ASG, OC, and LX and with 11% pN1 for OP and HP. There was 3% pN2 for LX, 4% pN2 for ASG and OC, and 6% pN2 for OP and HP. The DSS for patients with occult neck disease was statistically similar to that of N+ patients. Prognostic survival indicators included age, decade of treatment, T stage, N stage, TN stage, treatment modality, and recurrence. Patients over 65 years of age had poorer DSS than younger patients. Staging T, N, and TN affected survival at all sites. Local resection produced better DSS for ASG, OC, LX, and HP patients. Local resection with radiation therapy produced increased DSS for ASG and OC patients. There was no survival advantage for HP patients related to treatment modality. Treatment of the N0 neck with observation and later treatment for subsequent neck disease produced a survival advantage for patients in the ASG. This advantage was specific for ASG and LX patients staged T1N0. For patients staged T2N0, T3N0, and T4N0 at all four subsites there was no survival advantage for any of the four neck approaches. CONCLUSION: Lymph node metastasis significantly and negatively affects DSS in patients with SCCA of the OC, OP, LX and HP. The rate of occult neck disease (pN+) in N0 patients receiving meticulous workup is low. When present, it produces DSS rates similar to those found in N+ patients. In the study series, there was decreased survival in patients older than 65 years of age, in patients with advanced tumor (T, N, TN), and in patients with recurrent disease. None of the four current approaches to treatment of the N0 neck produces a significant survival advantage. Close observation with later treatment reserved for subsequent neck disease produces statistically similar survival (DSS) to the three elective (prophylactic) treatments and is a valid form of treatment. It may preclude unnecessary treatment of the neck with its attendant risks and complications. PMID- 15805873 TI - Audiologic testing and molecular analysis of 12S rRNA in patients receiving aminoglycosides. AB - BACKGROUND: Pathogenic mutations in the mitochondrial genome are associated with a wide variety of maternally inherited human diseases including sensorineural hearing loss (HL). A specific mutation, m.1555A>G in the mitochondrial 12S rRNA gene, is associated with predisposition to aminoglycoside ototoxicity and HL. Mutation screening in this gene has been recommended before use of aminoglycosides as a preventative strategy to reduce the risk of ototoxicity. OBJECTIVE: To study the incidence of mutations in the 12S rRNA gene in patients being treated with aminoglycosides and its correlation with ototoxicity. METHODS: Patients undergoing treatment with aminoglycosides were prospectively enrolled in this study (n = 27). Total dosage administered and therapeutic levels of the antibiotic were noted. All patients underwent high-frequency pure-tone audiometry pre- and posttherapy and sequencing of the 12S rRNA gene. In addition, 12S rRNA gene was also sequenced in 50 controls to characterize population specific polymorphisms. RESULTS: Five of 27 patients suffered from HL involving the high frequencies: four mild and one moderate. Only one of the five patients with ototoxicity harbored two sequence alterations in 12S rRNA of uncertain pathogenicity. The m.1555A>G and m.961delTInsCn mutations were not detected. CONCLUSIONS: High-frequency pure-tone audiometry is critical for detection of aminoglycoside-induced HL. In the Swiss population, screening for mutations in the 12S rRNA gene, before the initiation of aminoglycoside therapy, is not supported by this limited study. A larger multicenter and multicultural study is warranted to more definitively address this critical clinical issue. PMID- 15805874 TI - Positron-emission tomography for surveillance of head and neck cancer. AB - OBJECTIVES/HYPOTHESIS: To determine the diagnostic accuracy and the ideal timing of fluoro-fluorodeoxyglucose positron-emission tomography (PET) in the posttreatment surveillance of head and neck mucosal squamous cell carcinoma (HNSCC). STUDY DESIGN: Retrospective chart review. METHODS: Our sample includes 103 adult patients with 118 posttreatment PET scans who had undergone treatment for HNSCC. We correlated PET results with surgical pathology and clinical outcome in the subsequent 6 months. RESULTS: For the detection of locoregional persistent or recurrent HNSCC, PET scans had a sensitivity of 82%, specificity of 92%, positive predictive value (PPV) of 64%, negative predictive value (NPV) of 97%, and overall accuracy of 90%. For the detection of distant metastases, PET scans had a sensitivity of 89%, specificity of 97%, PPV of 85%, NPV of 98%, and overall accuracy of 96%. PET scans of the head and neck region performed greater than 1 month after the completion of radiation compared with scans performed within 1 month had a significantly higher sensitivity of 95% versus 55% (P < .01) and NPV of 99% versus 90% (P < .01). CONCLUSION: PET is effective in detecting distant metastases in the posttreatment surveillance for HNSCC patients. A negative PET is highly reliable for all sites. However, a positive PET in the head and neck region is unreliable because of a high false-positivity rate. PET of the head and neck region has a statistically significant risk of a false-negative reading when performed within 1 month of radiation. PMID- 15805875 TI - Inflammatory proteases in chronic otitis externa. AB - OBJECTIVES: Proteases are known to contribute to the pathogenesis of chronic inflammatory skin conditions such as atopic dermatitis and psoriasis. Inhibition of these proteases has shown promise in the treatment of these skin conditions. The purpose of this study was to measure the matrix metalloproteinases (MMP) and human neutrophil elastase (HNE) activities in chronic otitis externa (COE) and to determine whether administration of protease inhibitors recombinant alpha 1 antitrypsin (rAAT) and ilomastat might reduce these protease activities. STUDY DESIGN: Prospective and ex vivo. METHODS: Twenty-five ear canals with COE and 34 with no pathology (i.e., controls) were debrided and filled with saline. After a tragal pump and 1 to 2 minutes, the washes were collected and analyzed for MMP and HNE activities and the inhibitory activity of rAAT and ilomastat on these proteases, respectively. RESULTS: MMP and HNE levels were significantly higher (P = .0057 and .0112) in ears with COE than normal ears. MMP activity greater than 3 mAU/minute was observed in 30% of COE and 0% of controls (P = .0270). HNE activity greater than 3 mAU/minute was found in 77% of COE versus 7% of controls (P < .0001). Ilomastat and rAAT inhibited 60% of MMP and 98% of HNE activity, respectively, in COE ears. CONCLUSIONS: Elevated levels of proteases found in COE, MMP, and HNE may be inhibited with ilomastat and rAAT. The therapeutic potential of these protease inhibitors warrants investigation. PMID- 15805876 TI - Influence of aging and sex on voice parameters in patients with unilateral vocal cord paralysis. AB - OBJECTIVES: The objective of this study was to investigate influences of aging and sex on different voice parameters in patients with unilateral vocal cord paralysis (VCP). STUDY DESIGN: Retrospective review of patients with unilateral VCP. MATERIALS AND METHODS: Forty-seven patients, 22 males, 25 females (24-85 years), were enrolled in the study. The diagnosis of VCP was established by videolaryngostroboscopy. The acoustic parameters of jitter, shimmer, degree of subharmonic, noise to harmonic ratio, fundamental frequency, and maximal intensity were measured. The auditive voice analysis included roughness, breathiness, and hoarseness. Statistical analysis involves Pearson's bivariate correlation coefficients and two-way analysis of variance with interaction variables. RESOLUTION: After unilateral VCP in the elderly, some sex- and age related differences in the restriction of the voice can be documented. CONCLUSION: In general, the investigated voice parameters showed similar tendencies to those in otherwise healthy aging persons. PMID- 15805877 TI - Variables predicting distant metastases in thyroid cancer. AB - OBJECTIVES: Distant metastases from thyroid cancer are uncommon and have a variable prognosis. We present a series of patients with distant metastases to determine which patients are at risk of developing distant disease and to examine the significant prognostic variables. STUDY DESIGN: Retrospective chart review of 30 patients with distant metastases compared with 633 controls from the Mount Sinai Thyroid Cancer Database and literature review. METHODS: The prevalence of distant metastases was 4.5%, and median follow-up of survivors was 12.7 years. Histologic type was Hurthle cell carcinoma in 3, follicular in 3, papillary in 19, and 5 patients had focal anaplasia either in the primary site or regional metastases. Predictors for distant metastases, locoregional control, and survival were analyzed. RESULTS: Cumulative survival for patients with distant metastases was 49.5% at 10 years and 12.9% at 20 years. Site of metastases was lung in 26, bone in 11 and brain in 1 patient, with 8 patients having multiple sites. The median time to diagnosis of distant metastases was 3 months. Variables that predicted for development of distant disease were male sex, age, size, extrathyroidal extension, regional metastases, and elevated thyroglobulin. Survival in patients without distant disease was significantly better than those with distant metastases (P < .001). Variables that predicted poor outcome in patients with distant metastases on analysis were age greater than 45 years (P = .003) and histologic type of thyroid cancer (P = .009). CONCLUSION: Although patients with thyroid cancer and distant metastases may live prolonged periods with disease, it does significantly impact on patient survival. Age remains an important variable in both predicting for development of distant metastases and also influences long-term survival in patients with existing distant metastases. PMID- 15805878 TI - Comparison of diced cartilage graft wrapped in surgicell and diced cartilage graft wrapped in fascia: an experimental study. AB - HYPOTHESIS: This study was conducted to compare diced cartilage grafts wrapped in fascia and diced cartilage grafts wrapped in surgicell with respect to their resorption. STUDY DESIGN: Experimental study. METHODS: Eight male Wistar Albino rats were used. After general anesthesia, two different subcutaneous pockets (upper and lower pocket) were prepared in the rat's abdomen. Surgicell-wrapped homograft that was taken from the ear was placed into the upper subcutaneous pocket. Fascia-wrapped homograft which was taken from the ear was placed into the lower subcutaneous pocket. All the rats were killed at the end of 2 months, and the samples were collected to be examined histopathologically. Cartilage mass, foreign body reaction, fibrosis, chronic inflammation, and vascularization were researched. RESULTS: Diced cartilage wrapped surgicell stained negative for glial fibrillary acidic protein (GFAP). Diced cartilage wrapped in fascia stained positive for GFAP. There was a significant difference between two groups with respect to fibrosis, chronic inflammation, and cartilage mass. There was no significant difference between the two groups regarding vascularization. CONCLUSIONS: This study suggests that diced cartilage graft wrapped in surgicell was absorbed, whereas diced cartilage graft wrapped in fascia was not. PMID- 15805879 TI - Is word recognition correlated with the number of surviving spiral ganglion cells and electrode insertion depth in human subjects with cochlear implants? AB - OBJECTIVES/HYPOTHESIS: Speech perception scores using cochlear implants have ranged widely in all published series. The underlying determinants of success in word recognition are incompletely defined. Although it has been assumed that residual spiral ganglion cell population in the deaf ear may play a critical role, published data from temporal bone specimens from patients have not supported this hypothesis. The depth of insertion of a multichannel cochlear implant has also been suggested as a clinical variable that may be correlated with word recognition. In the current study these correlations were evaluated in 15 human subjects. STUDY DESIGN: Retrospective review of temporal bone histopathology. METHODS: Temporal bones were fixed and prepared for histological study by standard techniques. Specimens were then serially sectioned and reconstructed by two-dimensional methods. The spiral ganglion cells were counted, and the depth of insertion of the cochlear implant as measured from the round window was determined. Correlation analyses were then performed between the NU6 word scores and spiral ganglion cell counts and the depth of insertion. RESULTS: The segmental and total spiral ganglion cell counts were not significantly correlated (P > .50) with NU6 word scores for the 15 subjects. Statistically significant correlations were not achieved by separate analysis of implant types. Similarly, no significant correlation between the depth of insertion of the electrode array and postoperative NU6 word score was identified for the group. CONCLUSION: Although it is unlikely that the number of residual spiral ganglion cell counts is irrelevant to the determination of word recognition following cochlear implantation, there are, clearly, other clinical variables not yet identified that play an important role in determining success with cochlear implantation. PMID- 15805880 TI - Dental status and quality of life in long-term head and neck cancer survivors. AB - OBJECTIVES: To analyze how the dental status of long-term head and neck cancer survivors affects their subjective quality of life. STUDY DESIGN: Observational case series. METHODS: A convenience sample of 5-year head and neck cancer survivors underwent the following battery of tests: 1) targeted head and neck examination, 2) updated medical history, 3) dental evaluation, 4) standardized quality of life questionnaires. RESULTS: Eighty-six survivors were included in the study. The following associations were identified: 1) those who became edentulous secondary to cancer treatment and those without occlusion at time of the study demonstrated worse Pain, Activity, Recreation/Entertainment, Chewing, Swallowing, Speech, Eating in Public, Normalcy of Diet, Physical Well-Being, Social/Family Well-Being, Functional Well-Being, and Additional Concerns scores; 2) higher Decayed/Missing/Filled scores were associated with worse Pain, Disfigurement, Activity, Recreation/Entertainment, Employment, Chewing, Swallowing, Speech, Eating in Public, Understandability of Speech, Normalcy of Diet, Physical Well-Being, Additional Concerns scores, and weight loss; 3) decreased oral opening measurements were associated with worse Chewing, Swallowing, Eating in Public, Normalcy of Diet, Additional Concerns scores, and weight loss; 4) edentulous survivors who did not use dentures had worse Pain, Activity, Recreation/Entertainment, Understandability of Speech, and Eating in Public scores. CONCLUSIONS: Although previous studies have shown that many of the effects of cancer treatment disappear between 1 and 3 years, this study shows that the dental status has a persistent impact on subjective quality of life. PMID- 15805881 TI - Role of interleukins and transforming growth factor-beta in chronic rhinosinusitis and nasal polyposis. AB - OBJECTIVES: To determine the role of interleukin (IL)-4, IL-4 receptor (R), IL-6, IL-8, IL-11, and transforming growth factor (TGF)-beta in chronic rhinosinusitis (CRS) and chronic rhinosinusitis with nasal polyposis (CRS/NP). METHODS: Sinus tissue from patients undergoing endoscopic sinus surgery for CRS and CRS/NP was collected. Sinus tissue was then analyzed using reverse-transcription polymerase chain reaction (RT-PCR) to detect transcription of IL-4R, IL-6, IL-8, and IL-11. Sinus tissue samples were also cultured in vitro, treated with IL-4 for 24 hours, and real-time PCR was used to quantify the transcription of TGF-beta. RESULTS: Twenty patients were evaluated, 9 with CRS/NP and 11 with CRS alone. The mean age was 43 (20-74) years, with 13 females and 7 males. IL-4R, IL-6, IL-8, and IL-11 were identified by RT-PCR in all 20 patients. The transcription of TGF-beta was found to be 3.2 times greater in patients with CRS/NP than in patients with CRS alone (P = .047). CONCLUSION: IL-6, IL-8, and IL-11 are nonspecific markers of sinus inflammation being transcribed in patients with CRS and patients with CRS/NP. However, patients with CRS/NP demonstrate increased transcription of TGF beta in response to IL-4 treatment, suggesting an IL-4 mediated mechanism for stromal proliferation in the formation of nasal polyposis. PMID- 15805882 TI - Extraction of dental crowns from the airway: a multidisciplinary approach. AB - We describe two cases of airway foreign bodies (FB) consisting of a dental crown. The shape and composition of dental crowns complicate their extraction from the tracheobronchial tree, sometimes necessitating thoracotomy. We describe the use of a multidisciplinary approach involving rigid and flexible bronchoscopy in concert with the use of wire snares under fluoroscopic guidance for extraction of these challenging FB. These cases illustrate that this multidisciplinary approach can allow successful extraction of the difficult FB from much of the tracheobronchial tree and the avoidance of thoracotomy. PMID- 15805884 TI - An ex vivo model for reperfusion of laryngotracheal grafts. AB - OBJECTIVE: To describe the development of an ex vivo model to facilitate the study of ischemia-reperfusion injury in laryngotracheal grafts taken from Minnesota minipigs. STUDY DESIGN: This is a descriptive study. METHODS: Laryngotracheal grafts from Minnesota minipigs were harvested and placed in cold storage for 3 hours. Autologous blood was used to reperfuse the graft for up to 8 hours using the described reperfusion model. RESULTS: By altering retrieval technique and pressures within the graft, we demonstrated healthy grafts after 8 hours of reperfusion. CONCLUSIONS: Ex vivo reperfusion offers a range of experimental advantages over in vivo reperfusion including close control of interventions, ease of outcome measurement, and reduction in animal use. This model is now ready to be used for further studies of interventions to reduce ischemia-reperfusion injury in these grafts. PMID- 15805883 TI - Gene expression profiling in squamous cell carcinoma of the oral cavity shows abnormalities in several signaling pathways. AB - OBJECTIVES/HYPOTHESIS: To examine gene expression profiles in squamous cell carcinoma of the oral cavity (oral SCC) compared with histologically matched normal tissue. STUDY DESIGN: Fresh-frozen tissue was prospectively obtained from individuals undergoing surgical resections for oral SCC. METHODS: RNA was extracted from seven sets of oral SCC and matched normal tissue. Gene expression profiles were obtained by interrogation of Affymetrix Gene Chip Arrays with cRNA prepared from the tissue. Expression values were subjected to a paired t test. Genes that were judged to differ between oral SCC and normal tissue were annotated according to their name in the Affymetrix Netaffx database and according to their function as indicated by their Gene Ontology Consortium number. RESULTS: Of the 10,599 probe sets that were analyzed, 523 genes were abnormally expressed in SCC of the head and neck (P < or = .01), and 417 of these genes were abnormally expressed in all seven tumors in the same manner. Hierarchical clustering of the 121 genes that were abnormally expressed in cancerous relative to normal tissue (P < or = .001) showed that the tissue segregated into two groups consisting of normal and transformed tissue, as expected. The abnormal expression of two genes that were up regulated in oral SCC (ADAM 12 and PTHLH) and two genes that were down-regulated in SCCHN (EMP-1 and P11) were confirmed by real-time polymerase chain reaction and immunohistochemistry (ADAM 12) using additional sets of tissue. CONCLUSIONS: The data showed that oral SCC aberrantly express genes in cellular pathways related to proliferation, apoptosis, extracellular matrix degradation, adhesion, transforming growth factor-beta signaling, and transcription. Further work is needed to determine the role of these genes in the development and progression of oral SCC. PMID- 15805885 TI - Headache: a quality of life analysis in a cohort of 1,657 patients undergoing acoustic neuroma surgery, results from the acoustic neuroma association. AB - OBJECTIVES: On the basis of survey results of the Acoustic Neuroma Association (ANA), we report patient ratings of postoperative headache (POH) symptoms, determine its effect on quality of life (QOL), and review the literature regarding POH after acoustic neuroma (AN) treatment. STUDY DESIGN: In this cohort study, 1,657 patients who underwent surgical treatment of AN reported their experiences of POH. METHODS: A detailed questionnaire was mailed to members of the ANA to identify preoperative and postoperative headache symptoms, complications, and long-term effects on physical and psychosocial function. Questions were answered by 1657 (85.4%) respondents that were intended to qualify and quantify the effects of POH, including QOL issues. Responses were analyzed by tumor size, surgical approach, and patient age and sex. Statistical analysis was performed with the SPSS software. RESULTS: Preoperative headache was reported in approximately one third of respondents. Typical POHs occurred more than once daily (46%), lasted 1 to 4 hours in duration (43.1%), and were of moderate intensity (62.6%). The worst headaches were rated as "severe" by 77% of respondents. Treatment most often reported for typical headaches were nonprescription medications including nonsteroidal anti-inflammatory drugs in 61.3% (P < .01) and regular use of narcotics in 15%. Patients who underwent the retrosigmoid approach were significantly more likely to report their worst POH as "severe" (82.3%) compared with the translabyrinthine (75.2%) and middle fossa approaches (63.3%). Women and younger patients tended to have poorer outcomes with regard to POHs. CONCLUSIONS: In this large cohort study of AN patients, POH was a significant morbidity among AN patients with persistent headaches. Treating physicians should be aware of the risk factors identified and the effect POH has on the QOL when counseling patients regarding optimal treatment management. PMID- 15805886 TI - Nerve monitoring and stimulation during endoscopic neck surgery in the pig. AB - OBJECTIVES: To determine the feasibility of recurrent laryngeal nerve monitoring and stimulation during endoscopic neck surgery in an animal model. STUDY DESIGN: Prospective, nonrandomized experimental investigation in a porcine model. METHODS: Bilateral recurrent laryngeal nerve monitoring and stimulation was accomplished during endoscopic neck surgery in five domestic pigs. Each pig was intubated with an electromyography endotracheal tube. Recurrent laryngeal nerve function was monitored throughout the endoscopic neck surgery with a nerve integrity monitor system. An endoscopic surgical pocket was created in the neck using blunt dissection followed by low-pressure carbon dioxide insufflation. Under direct endoscopic visualization, the trachea, thyroid gland, and associated vasculature were identified. The recurrent laryngeal nerve was identified on each side of the animal and was successfully stimulated with a monopolar stimulator probe. RESULTS: Ten of ten recurrent laryngeal nerves were successfully monitored and stimulated. No significant complications were encountered during the procedures. CONCLUSIONS: Recurrent laryngeal nerve monitoring and stimulation may be successfully accomplished during endoscopic neck surgery. PMID- 15805887 TI - Fat obliteration in paranasal sinuses: a comparative magnetic resonance imaging and histopathologic study. AB - OBJECTIVES: To assess postoperative changes after fat tissue obliteration of the paranasal sinuses with the use of magnetic resonance imaging (MRI) and correlate the findings with correspondent histology. STUDY DESIGN: By using an animal model with fat obliteration of the maxillary paranasal sinus. METHODS: We correlated postoperative changes of the fatty tissues by means of histopathologic analysis and MRI. The study group included 15 rabbits undergoing autologous fat tissue obliteration of their maxillary paranasal sinus. After 1 month (n = 5), 3 months (n = 5), and 6 months (n = 5), both MRI and histopathologic evaluations of the fatty tissue status were performed. Contrast enhanced MRI was used to identify vital fat tissue. Subsequently, MRI findings were compared with a correspondent histologic status and proliferative factors such as angio- and osteogenesis and presence of abundant granulocytes, macrophages, and giant cells. RESULTS: After a period of 6 months, the obliteration sites in all animals showed vital fat tissues, whereas at 1 month after surgery, vital fat tissue was rarely observed. The microscopic appearance of the obliteration tissue after 1 month was characterized by fat tissue necrosis and distinct tissue reactions including blood vessel dilatation, abundant macrophages, granulocytes, and lymphocytes. MRI after 1 month showed a clear contrast enhancement because of the hyperemia and inflammation reaction. CONCLUSION: Fat tissue transplants used for obliteration of paranasal sinuses are almost completely degraded after transplantation and replaced by vital fat tissue over a period of at least 6 months. Contrast enhanced MRI is a well-suited technique for follow-up imaging and assessing the transplant vascularization and tissue remodeling status. PMID- 15805888 TI - Anterior vocal commissure invasion in laryngeal carcinoma diagnosis. AB - BACKGROUND: Laryngeal carcinoma involving anterior vocal commissure (AVC) represents a great challenge for staging and treatment. OBJECTIVES: To compare laryngoscopy and computed tomography (CT) scan efficiency in staging tumors extending to the AVC. We also analyzed the helicoidal axial CT scan accuracy in recognizing this larynx subregion invasion. MATERIAL AND METHODS: Fifty-two glottic and supraglottic laryngeal squamous cell carcinoma patients with tumoral extension to the AVC were prospectively studied from August 2001 to August 2003 at the National Cancer Institute (Rio de Janeiro, Brazil). All patients underwent videolaryngoscopic examination and direct laryngoscopy for lesion extension analysis and biopsy. After AVC helicoidal axial CT scan with sagittal and coronal 1.0 mm thick reconstruction, patients were submitted to surgical treatment. The same pathologist analyzed all surgical specimens. RESULTS: When compared with pathologic stage, clinical endoscopic classification was correct in 40.38% of cases (40% for T1, 29.41% for T2, 46.43% for T3, and 50% in T4). Helicoidal axial CT scan accuracy for AVC tumors was 75% (P = .0001), being more important for T2 (62.50%), T3 (73.91%), and T4 (88.24%) lesions. Identification of radiologic signs described as gross radiologic anterior commissure involvement (GRACI) increased radiologic image staging accuracy to over 96%. CONCLUSIONS: Endoscopic evaluation understaged tumors in all clinical stages but really T1. Helicoidal axial CT scan reformatted to 1.0 mm thick played an important role in correctly staging more advanced AVC laryngeal tumors. Radiologic signs, here identified as GRACI, may be very helpful for tomographic staging and patient treatment. PMID- 15805889 TI - Effect of adenotonsillectomy on middle ear status in children. AB - OBJECTIVE: To determine the effects of adenotonsillectomy as compared with watchful waiting on the middle ear status of children. STUDY DESIGN: Randomized controlled trial. METHODS: We recruited 300 children between 2 and 8 years of age who were selected for adenotonsillectomy according to current medical practice. Excluded from the trial were children with very frequent throat infections (more than 6 per year) or obstructive sleep apnea. Participants were randomly assigned to either adenotonsillectomy or watchful waiting. Main outcome measure was the percentage of children with unilateral or bilateral otitis media diagnosed at the scheduled follow-up visits according to an algorithm combining tympanometry and otoscopy. RESULTS: The percentages of children in the adenotonsillectomy and watchful waiting group diagnosed with otitis media at baseline and at 3, 6, 12, 18, and 24 months were 27.7 versus 30.5, 16.8 versus 25.2, 18.3 versus 21.2, 12.3 versus 15.2, 17.6 versus 15.5, and 14.7 versus 10.3%, respectively (P < .10). In the subgroup of children selected for adenotonsillectomy predominantly because of recurrent or persistent otitis media, hearing loss, or recurrent upper respiratory tract infections (n = 111) and in the subgroup of children diagnosed with otitis media at inclusion (n = 82), the occurrence of otitis media did not differ significantly between the adenotonsillectomy and watchful waiting group during the entire follow-up period. CONCLUSION: We conclude that in a large proportion of children selected for adenotonsillectomy according to current medical practice, including those with otitis media or related complaints, no beneficial effect of adenotonsillectomy on middle ear status is to be expected. PMID- 15805890 TI - Sleep endoscopy versus modified Mallampati score in sleep apnea and snoring. AB - OBJECTIVES/HYPOTHESIS: The aim of this study is to quantify the correlation between a large tongue classified by the modified Mallampati score (MMS) and obstruction at the tongue base observed during sleep endoscopy in patients with socially unacceptable snoring (SUS) and obstructive sleep apnea syndrome (OSAS). STUDY DESIGN: Prospective. METHODS: One hundred twenty-seven patients with obstructive sleep apnea and SUS underwent sleep endoscopy. Patients were classified into four specific groups according to their MMS. After intravenous administration of midazolam, obstructive events in the upper airway were classified. The level of obstruction was expressed as palatal (2), palatal and to a lesser degree base of the tongue/laryngeal level (2), tongue base/laryngeal level and to a lesser degree palatal (3), and tongue base/laryngeal level (3). The degree of correlation between the level of obstruction and the MMS was analyzed by means of Spearman's rank-order correlation. RESULTS: No linear association between the level of obstruction and MMS could be found. A large tongue (MMS 3 or 4) does not predict obstruction at the base of the tongue. Patients classified as MMS 1 or 2 did not frequently show obstruction at the base of the tongue (9 of 38, 23,6%). The majority of the population showed retropalatal obstruction. A large tongue (MMS 3 or 4) was often accompanied by retropalatal obstruction, 76% of the cases (64 of 89). CONCLUSIONS: No positive correlation exists between a large tongue and obstruction at tongue base level. In both groups, retropalatal obstruction occurred more often than retrolingual obstruction. Sleep endoscopy provides qualitative insight into important changes during sleep in patients with SUS and OSAS. In addition, anatomic aberrations are clearly visualized in a dynamic setting. Classification of the size of the tongue, as assessed by the MMS, may be of great importance as well. PMID- 15805891 TI - Hyoidthyroidpexia: a surgical treatment for sleep apnea syndrome. AB - OBJECTIVES/HYPOTHESIS: The aim of this study is to evaluate the results of primary hyoidthyroidpexia (HTP) and HTP after previous uvulopalatopharyngoplasty (UPPP) in patients with obstructive sleep apnea syndrome (OSAS). STUDY DESIGN: Prospective case series. METHODS: Thirty-one patients with obstruction at tongue base level and moderate to severe sleep apnea syndrome underwent HTP. Seventeen patients underwent surgery after an unsuccessful UPPP (secondary HTP), and in 14 patients, primary HTP was performed. RESULTS: Patients who underwent primary HTP showed a significant decrease in apnea hypopnea index (AHI) (P = .007), whereas those patients who had secondary HTP did not (P = .06). Overall, the AHI significantly changed (P = .0005). Visual analogue scales for snoring and hypersomnolence and the Epworth sleepiness scores showed significant improvement for both groups, without any difference between them. HTP was considerably less painful when compared with UPPP. CONCLUSION: This study demonstrates that HTP, in particular as primary treatment in cases of obstruction at tongue base level, is a valuable addition to the therapeutic armamentarium of moderate to severe OSAS. Selection criteria are moderate to severe OSAS with preferably a body mass index less than 27, multilevel obstruction with emphasis on the base of tongue, small tonsils, and normal uvula, without a floppy epiglottis or a palatal stenosis after UPPP. PMID- 15805892 TI - Sodium morrhuate sclerotherapy for the treatment of benign lymphoepithelial cysts of the parotid gland in the HIV patient. PMID- 15805893 TI - Conservative transoral removal of hilar submandibular salivary calculi. PMID- 15805894 TI - Use of image-guided computed tomography-magnetic resonance fusion for complex endoscopic sinus and skull base surgery. PMID- 15805896 TI - Beta-trace protein: an unknown marker for cerebrospinal fluid leaks. PMID- 15805898 TI - Questioning the relationship between cochlear otosclerosis and sensorineural hearing loss. PMID- 15805899 TI - Optical coherence tomography of choroidal nevus in 120 patients. AB - OBJECTIVE: To describe the optical coherence tomography (OCT) findings of choroidal nevi. METHODS: Retrospective, single-center case series of 120 eyes of 120 consecutive patients with choroidal nevi who were evaluated by OCT. Diagnostic imaging was performed with a Zeiss StratusOCT Model 3000 (Carl Zeiss Ophthalmic Systems, Dublin, CA) using scan acquisition protocols of 6 radial lines and retinal thickness analysis overlying the nevus. RESULTS: The mean patient age was 59 years (median, 60 years; range, 14-87 years). The choroidal nevus was a mean of 5.2 mm in basal dimension and 1.7 mm in thickness and was located a mean of 2.7 mm from the optic disk and 2.5 mm from the foveola. Related retinal findings by ophthalmoscopic evaluation included overlying retina edema (3%), subretinal fluid (16%), retinal thinning (0%), drusen (58%), and retinal pigment epithelium (RPE) detachment (2%). In comparison, related retinal findings at the site of the nevus by OCT included overlying retina edema (15%), subretinal fluid (26%), retinal thinning (22%), drusen (41%), and RPE detachment (12%). Furthermore, OCT permitted classification of the overlying retinal edema as focal cystoid (3%), diffuse cystoid (8%), coalescent cystoid (3%), and noncystoid edema (1%). By OCT, the overlying retina was normal thickness (32%), thinned (22%), or thickened (45%), and photoreceptor loss or attenuation was noted in 51% of cases. Specific OCT findings of the choroidal nevus were limited to its anterior surface with minimal penetration into the mass. These findings included increased thickness of the RPE/choriocapillaris layer (68%) and optical qualities within the anterior portion of the nevus of hyporeflectivity (62%), isoreflectivity (29%), and hyperreflectivity (9%). Hyporeflectivity was observed in 68% of pigmented nevi and 18% of nonpigmented nevi. When comparing OCT with clinical examination, OCT was more sensitive in the detection of related retinal edema, subretinal fluid, retinal thinning, photoreceptor attenuation, and RPE detachment. CONCLUSIONS: OCT is a useful diagnostic modality for imaging the retina overlying a choroidal nevus. Numerous overlying changes such as subretinal fluid, retinal edema, retinal thinning, and photoreceptor attenuation are visible by OCT. PMID- 15805900 TI - Guidelines for interpreting retinal photographs and coding findings in the Submacular Surgery Trials (SST): SST report no. 8. AB - PURPOSE: To describe the guidelines followed by the Submacular Surgery Trials (SST) Research Group in the interpretation of color fundus photographs and fluorescein angiograms of subfoveal choroidal neovascular lesions evaluated in the SST and to assist ophthalmologists in applying the results of the SST. METHODS: Stereoscopic color fundus photographs and fluorescein angiograms of the study eye and nonstudy eye of 1,015 patients with subfoveal choroidal neovascular lesions secondary to age-related macular degeneration, ocular histoplasmosis syndrome, or idiopathic choroidal neovascularization (CNV) were obtained and graded by certified SST fundus photograph readers at the baseline examination in three randomized clinical trials comparing surgery with observation. Adherence to the inclusion and exclusion criteria and ocular features that might affect visual outcome were documented. Stereoscopic color fundus photography and fluorescein angiography were repeated 1 month after randomization for patients assigned to surgery to provide documentation that surgery was performed and to assess compliance with the surgery protocol. Photographs and fluorescein angiograms of both the study eye and the fellow eye in all patients then were obtained 3 months, 6 months, and 12 months after randomization and then annually up to 48 months. The kappa statistic was used to evaluate interobserver reliability of photograph gradings. RESULTS: Lesion components at baseline included classic CNV, occult CNV, and features contiguous to CNV, including blood, fibrous tissue, hypofluorescence not corresponding to blood, serous detachment of the retinal pigment epithelium, and prior areas of laser photocoagulation. At follow-up, fluorescein leakage from CNV was assessed peripheral to or within the area of the retinal pigment epithelium abnormality after surgery. The lesion at follow-up could include any of the features identified at baseline as well as retinal pigment epithelium abnormalities, such as mottling of the retinal pigment epithelium with a subtle transition to normal retinal pigment epithelium or a very sharply demarcated, markedly hypopigmented area that was easily distinguished from the surrounding retinal pigment epithelium. kappa statistics for interobserver reliability ranged from good (0.47) to excellent (1.00) for features graded at baseline and follow-up. CONCLUSIONS: Although some of the definitions essential to the interpretation of the SST are similar to those used in the Macular Photocoagulation Study and randomized clinical trials of photodynamic therapy with verteporfin, this guideline provides new information regarding lesion components at baseline as well as standardized descriptions of lesions after submacular surgery. These descriptions from the SST assist in understanding what lesions were studied, when additional treatment was considered after surgery, and how anatomical results should be interpreted. PMID- 15805901 TI - Microbiologic studies of multiple-dose containers of triamcinolone acetonide and lidocaine hydrochloride. AB - PURPOSE: Reports of high endophthalmitis rates following intravitreal triamcinolone acetonide (TA) injection have raised concerns about safety. One possible risk factor is the use of multiple-dose containers. The purpose of this study was to determine whether there is a substantial risk of contamination of multiple-dose containers. METHODS: Microbiologic Challenge Study: Suspensions of a known quantity of several microorganisms were placed in samples of TA to determine whether the preservative, benzyl alcohol 0.99%, is rapidly bacteriocidal. Simulated Clinical Use Study: After ten days of simulated clinical use of multiple-dose containers of TA and lidocaine hydrochloride 4%, samples were cultured. Additionally, samples from lidocaine containers in actual clinical use were cultured. RESULTS: Microbiologic Challenge Study: Four of five challenge organisms demonstrated moderate growth, even after 24 hours of exposure to the benzyl alcohol preservative. Simulated Clinical Use Study: Simulated clinical use of multiple-dose containers of TA and lidocaine did not result in contamination, nor did actual clinical use of multiple-dose lidocaine containers. CONCLUSIONS: No culture-proven contamination of multiple-dose containers in simulated or actual clinical use was found. However, TA suspension is capable of harboring viable bacterial and fungal pathogens. The use of multiple-dose containers of triamcinolone for intravitreal injections is therefore discouraged. PMID- 15805902 TI - Transvitreal limited arteriovenous-crossing manipulation without vitrectomy for complicated branch retinal vein occlusion using 25-gauge instrumentation. AB - PURPOSE: To evaluate a new technique, 25-gauge transvitreal limited arteriovenous crossing manipulation without vitrectomy (LAM), for the treatment of branch retinal vein occlusion (BRVO) complicated by macular hemorrhage and/or macular edema recalcitrant to grid laser photocoagulation. METHODS: Twelve eyes of 12 patients underwent LAM for BRVO performed by a single surgeon (M.S.H.) using the 25-gauge nitinol flexible-extendable blunt pick. The presence or absence of intraoperative reperfusion visualization, pre- and postoperative visual acuity, macular thickness as measured by optical coherence tomography, intraocular pressure, and lens status were evaluated. RESULTS: Restoration of blood flow was noted in all patients and was based on intraoperative reestablishment of a red column of erythrocytes through the previously closed vessel. Mean visual acuity improved from 20/200 (logarithm of the minimal angle of resolution [LogMAR] +/- SD, 1.00 +/- 0.32) preoperatively to 20/70 (LogMAR +/- SD, 0.56 +/- 0.28) (P = 0.0003) at the final visit. Eleven (92%) of 12 eyes had >or=2 lines of visual improvement. Five eyes (45%) had final visual acuity of 20/50 or better. Mean macular thickness +/- SD improved from 401.0 +/- 73.2 to 178.7 +/- 19.6 microm (P < 0.0001) at the final visit. No statistically significant difference was noted in cataract progression or intraocular pressure. Mean follow-up +/- SD was 49.9 +/- 19.6 weeks. All patients were observed for at least 12 weeks. CONCLUSION: LAM may achieve outcomes comparable with those of arteriovenous adventitial sheathotomy for complicated BRVO. PMID- 15805903 TI - Incidence of sclerotomy-related retinal breaks. AB - PURPOSE: To estimate the incidence of sclerotomy-related retinal breaks (SRRBs) and identify possible associated risk factors. METHODS: Medical records of patients who underwent pars plana vitrectomy (PPV) from January 1994 to January 1999 at the King Khaled Eye Specialist Hospital in Riyadh, Saudi Arabia, were reviewed. The diagnosis, procedure, and possible associated risk factors were documented, in addition to the presence of SRRBs and time of discovery. RESULTS: Of the 441 eyes, 3.1% developed early SRRBs, while 2.9% of 350 eyes developed late SRRBs. Most of the eyes that developed SRRBs were phakic, and penetrating trauma was found statistically to increase the risk of SRRBs. CONCLUSIONS: SRRBs are an uncommon complication of PPV. Early detection and proper management can prevent serious sequelae of such complications. PMID- 15805904 TI - The macular buckling procedure in the treatment of retinal detachment in highly myopic eyes with macular hole and posterior staphyloma: mean follow-up of 15 years. AB - PURPOSE: To evaluate the long-term anatomical and functional results of the macular buckling procedure in myopic macular hole with retinal detachment and posterior staphyloma. METHODS: Twenty-five consecutive highly myopic eyes with retinal detachment, macular hole, and posterior staphyloma, which were operated between February 1983 and April 1992, were retrospectively studied. The last examination was in June 2002. Follow-up of the 25 patients ranged from 10 years to 19 years (mean, 15 years). Macular buckling with an episcleral sponge was the initial and only procedure. In all operated eyes, anchoring of the sponge was away from the posterior pole. Best-corrected visual acuity was evaluated before and after surgery. The axial length of the eyeball was also measured by A-scan ultrasonography before treatment. RESULTS: Twenty-two of 25 eyes were successfully treated with a single procedure. In the remaining three eyes, retinal reattachment was obtained after a second buckling operation, which was considered necessary due to loosing of the sponge fixation sutures in two cases and the location of the hole beside the buckle in one case. The mean overall visual acuity was improved after treatment. CONCLUSIONS: The macular buckling procedure with anchoring of the sponge away from the posterior segment provided efficient scleral indentation of long duration for closure of the macular hole and retinal reattachment. PMID- 15805905 TI - Removal of adherent cortical vitreous plaques without removing the internal limiting membrane in the repair of macular detachments in highly myopic eyes. AB - PURPOSE: To report anatomic and visual improvement after vitrectomy with removal of adherent plaques of vitreous and gas tamponade in highly myopic eyes with detachments over posterior staphylomata. METHODS: Eyes with retinal detachment over staphylomata were treated with vitrectomy and vitreous cortex removal. Adherent plaques of vitreous were invariably found on the detached retina and were easier to identify with the aid of intraocular triamcinolone. Removal of the vitreous plaques and placement of long-acting gas affected surgical repair. Outcome measures included best-corrected visual acuity and retina reattachment confirmed by biomicroscopic appearance and optical coherence tomography findings. RESULTS: Six eyes of five consecutive patients were treated. The mean preoperative best-corrected visual acuity was 20/100, and the mean time of documented detachment was 21 months. The internal limiting membrane was not removed in any patient. After surgery, all patients' retinas remained attached during the mean follow-up period of 19.1 months. At the end of the follow-up period, the mean best-corrected visual acuity was 20/60, an improvement that was statistically significant (P = 0.027). CONCLUSION: Retinal detachment over staphylomata in highly myopic eyes appears to be tractional from unresolved forces caused by adherent plaques of vitreous. Removal of the vitreous, without removing the internal limiting membrane, can reattach the retina and lead to anatomic and visual acuity improvements. PMID- 15805906 TI - Results of a multicenter clinical trial to evaluate the preferential hyperacuity perimeter for detection of age-related macular degeneration. AB - PURPOSE: To compare the preferential hyperacuity perimeter (PHP) with an Amsler grid in detection of age-related macular degeneration (AMD). METHODS: Patients underwent refraction, visual acuity examination, PHP, Amsler grid examination, and macular photography. RESULTS: One hundred fifty patients participated in the trial. Of 19 eyes with neovascular AMD, 19 (100%) were positive on the PHP, and 10 (53%), on the Amsler grid. Of 27 eyes with geographic atrophy, 26 (96%) were positive on the PHP, and 12 (44%), on the Amsler grid. Of 20 eyes with intermediate AMD, 14 (70%) were positive on the PHP, and 4 (20%), on the Amsler grid. Of 51 eyes with early AMD, 21 (41%) were positive on the PHP, and 4 (8%), on the Amsler grid. Of 33 eyes with no AMD, 6 (18%) were positive on the PHP, and none, on the Amsler grid. Thus, 80 (68%) of 117 patients with AMD had a positive PHP, while 30 (26%) had positive results of Amsler grid examination (P < 0.001, McNemar test). CONCLUSION: The PHP had greater sensitivity, although with a relatively high rate of false-positive results for healthy individuals, than the Amsler grid in detecting AMD-related lesions. PMID- 15805907 TI - Bilateral idiopathic multifocal retinal pigment epithelium detachments in otherwise healthy middle-aged adults: a clinicopathologic study. AB - PURPOSE: To present the results of a clinicopathologic study of patients with bilateral idiopathic multifocal serous retinal pigment epithelial detachments (RPEDs). METHOD: Retrospective review of the medical records of three middle-aged patients with innumerable RPEDs. The eye of one patient was available for histopathologic examination. RESULTS: The presence of multiple serous RPEDs was verified with fluorescein angiography and optical coherence tomography in two patients and by histopathologic examination of one eye of a third patient, who developed a submacular hematoma and vitreous hemorrhage caused by a focal area of choroidal neovascularization. CONCLUSIONS: No cause for the multifocal serous RPEDs was found. PMID- 15805908 TI - Histopathologic evaluation of the internal limiting membrane surgically excised from eyes with diabetic maculopathy. AB - PURPOSE: To histopathologically evaluate the internal limiting membrane (ILM) in diabetic eyes with macular edema as compared to nondiabetic controls. METHODS: The authors ultrastructurally and immunohistochemically studied ILM specimens that were intentionally peeled from five eyes with diabetic maculopathy, comprising four with diffuse diabetic macular edema and one with macular hole accompanying diabetic retinopathy (DM group), and five with nondiabetic idiopathic macular hole (MH group). They compared ultrastructural and immunohistochemical findings between the two groups. RESULTS: A larger amount of cellular elements was observed on the vitreous side of the ILM in the DM group. The thickness of the ILM in the DM group was significantly increased (mean 4.8 +/ 1.6 microm) compared with that in the MH group (1.8 +/- 0.6 microm) (P < 0.0001). Immunoreactions for heparan sulfate proteoglycan in the ILM were more abundant in the DM group than in the MH group. CONCLUSION: The ILM thickening and cell abundance on the vitreous surface might contribute to the course and the pathogenesis of diabetic maculopathy. PMID- 15805909 TI - Associations between specific measures of vision and vision-related quality of life in patients with bothnia dystrophy, a defined type of retinitis pigmentosa. AB - PURPOSE: To assess the relationship between objective tests of visual function and vision-related quality of life in patients with Bothnia dystrophy (BD), a retinal dystrophy of retinitis pigmentosa type with progressive maculopathy. METHODS: Forty-nine patients were tested. Weighted distance logMAR visual acuity (WVA), weighted logMAR low contrast VA (WCS), and binocular visual field (VF) areas were calculated. Vision-related quality of life (VRQL) was assessed using the 25-item National Eye Institute-Visual Function Questionnaire (NEI-VFQ-25). Correlation statistics were used and adjusted analyses of the relationship between the composite score and the objective visual function tests were performed with multiple linear regression. RESULTS: VRQL was significantly correlated with age, WVA, WCS, and binocular VF areas (P < 0.001). Calculation of partial correlation coefficients showed age to be significantly correlated only with VF (V-4-e) area (P < 0.0001). Multiple linear regression analyses revealed age and WVA to be significantly associated with the NEI-VFQ-25 composite score (P < 0.02 and P = 0.001, respectively). WVA alone was the strongest predictor of self-reported experience of total visual function in BD patients (r 2= 0.69). CONCLUSIONS: A strong relationship between objective tests of visual function and patient perceived VRQL as assessed by a questionnaire was found. WVA was the strongest predictor and together with age explained almost 70% of the variability of the composite score of the questionnaire. PMID- 15805910 TI - Prevalence of patients presenting with neovascular age-related macular degeneration in an urban population. AB - PURPOSE: To determine the number and type of new cases of neovascular age-related macular degeneration (AMD) present in a defined urban population and to establish the proportion that would be recommended for treatment with verteporfin or laser photocoagulation. METHODS: Patients referred to an ophthalmic center in Vienna during a 10-week period because of recent deterioration in vision caused by newly diagnosed neovascular AMD were included. RESULTS: Neovascular AMD was diagnosed in 168 eyes in 153 patients. One hundred one eyes (60.1%) had lesions that were occult with no classic choroidal neovascularization (CNV); of these, 70 were subfoveal, 19 were juxtafoveal, and 12 were extrafoveal. Thirty-five eyes (20.8%) had predominantly classic lesions; of these, 27 were subfoveal, 6 were juxtafoveal, and 2 were extrafoveal. Thirty-two eyes (19.0%) had minimally classic lesions, of which 31 were subfoveal and 1 was extrafoveal. In accordance with consensus guidelines from a panel of experts and with American Academy of Ophthalmology's Preferred Practice Pattern guidelines, 33 lesions (17%) would be considered for treatment with verteporfin therapy. A further 37 subfoveal lesions with occult with no classic CNV and 7 juxtafoveal lesions with occult with no classic CNV might also benefit from verteporfin therapy if there is evidence of presumed recent disease progression. Five lesions (3.0%) could have been treated with laser photocoagulation according to Macular Photocoagulation Study criteria. CONCLUSIONS: These results suggest that verteporfin therapy substantially increases the number of patients with treatable neovascular AMD. PMID- 15805911 TI - Fourier transform infrared and 1h nuclear magnetic resonance spectroscopic findings of silicone oil removed from eyes and the relationship of emulsification with retinotomy and glaucoma. AB - PURPOSE: To investigate whether structural changes occur in silicone oil that has been left in the eye for different periods and to research the relationship of emulsification with the time during which silicone oil is left in the eye, glaucoma, and wide relaxing retinotomy. METHODS: Samples of silicone oil were randomly removed from the eyes of 22 patients. Gel permeation chromatography (GPC) and spectroscopy were used to identify any changes in the silicone oil. The relationship between spectroscopically determined emulsification with time spent in the eye, glaucoma, and wide relaxing retinotomy was investigated statistically. RESULTS: GPC revealed that no change was found in the 1,000 centistoke silicone oil model after it had been removed from the eye. Spectroscopy identified absorption peaks belonging to water in addition to those belonging to the functional groups. The relationships between emulsification and time spent in the eye and glaucoma were found to be statistically insignificant, while that between emulsification and wide relaxing retinotomy was statistically significant. CONCLUSION: Intravitreally applied silicone oil undergoes no structural change. Wide relaxing retinotomy constitutes a risk factor for silicone oil emulsification, which can occur earlier than expected and before it can be observed clinically. PMID- 15805912 TI - 2,4-Dinitrophenol pharmacologically promotes retinal detachment in rabbits. AB - PURPOSE: The most difficult and unpredictable step of macular translocation surgery is creating the retinal detachment. The authors evaluated the efficacy of 2,4-dinitrophenol (2,4-DNP) to promote retinal detachment in the rabbit. METHODS: A vitrectomy was performed in each eye of a Dutch-belted rabbit. One eye was injected with 0.1 cc of a 5 mmol/L 2,4-DNP, the other eye with 0.1 cc of BSS+. After 30 minutes, the minimum aspiration pressure required to visibly elevate the retina was recorded. Four nonvitrectomized eyes received an intravitreal injection of either 0.1 cc of BSS+ or 5 mmol/L 2,4-DNP, and were enucleated and fixated for histopathologic examination. RESULTS: In the 12 masked eyes, the mean aspiration pressure decreased from 217 +/- 20 mmHg in the six BSS+ eyes to 117 +/ 20 mmHg in the six 2,4-DNP treated eyes (P = 0.0022). A retinal detachment was present in three of six masked and two of two unmasked 2,4-DNP treated eyes and none of eight BSS+ treated eyes. There was no short-term toxicity to the retina at the light microscope level. CONCLUSION: Intravitreal injection of 2,4-DNP reduced the retinal adhesive force by over 50% when compared to the BSS+ treated control eyes, without any short-term retinal toxicity. PMID- 15805913 TI - Study of inhibitory effects of an antiangiogenic somatostatin-camptothecin conjugate on laser-induced choroidal neovascularization in rats. AB - PURPOSE: To evaluate the ocular toxicity and efficacy of intravitreal camptothecin-somatostatin conjugate (JF-10-81), a somatostatin type 2 receptor directed, antiangiogenic compound. METHODS: Part 1: New Zealand albino rabbits (except for controls) were injected intravitreally with conjugate at various concentrations. Preoperative and postoperative ocular examinations and electroretinography were performed until animals were killed for histology. Part 2: Long-Evans pigmented rats had choroidal neovascularization (CNV) induced by argon laser. One eye per animal was injected with concentration 10M (safe dose), whereas the other eyes were controls and received 30 microL of sterile water at different time intervals after laser application. Fluorescein angiography was performed at various time points to evaluate the lesions and confirm presence of CNV. Animals were euthanized. The eyes were immediately enucleated and prepared for histologic examination. RESULTS: Part 1: No clinical changes were seen in groups receiving 10(-8)M, 10(-7)M, 10(-6)M, and 10(-5) M of conjugate. Electroretinography showed decreasing b-wave amplitude in groups receiving 10(-4) M and 10(-3) M; cataracts also developed in these eyes. Part 2: Fluorescein angiography revealed that intravitreal injection of somatostatin conjugate JF-10 81 favorably affected the development of CNV when the treatment was performed at least 1 week after the laser application. These results were statistically significant. Histologic analysis results of eyes treated 2 weeks after laser application also showed significant benefit. CONCLUSIONS: Part 1: Camptothecin somatostatin conjugate injected intravitreally appeared safe at concentrations of 10(-5)M or less. Part 2: Conjugate JF-10-81 at a concentration of 10(-5)M administered intravitreally 1 to 3 weeks after laser demonstrated statistically significant efficacy in the treatment of choroidal neovascularization. PMID- 15805914 TI - Diagnostic and therapeutic challenges. PMID- 15805915 TI - Serous detachment of macula in Behcet disease. PMID- 15805916 TI - Acute posterior multifocal placoid pigment epitheliopathy and choroidal neovascularization. PMID- 15805917 TI - Necrotic melanocytoma of the optic disk with central retinal vascular obstruction. PMID- 15805918 TI - Choroidal metastasis from lung cancer masquerading as sarcoidosis. PMID- 15805919 TI - Isolated, bilateral intraocular lymphoma in a 15-year-old girl. PMID- 15805920 TI - Foveal schisis as a cause of retinal detachment secondary to macular hole in juvenile X-linked retinoschisis. PMID- 15805921 TI - Photodynamic therapy for choroidal neovascularization secondary to sympathetic ophthalmia. PMID- 15805922 TI - Choroidal neovascularization in sorsby fundus dystrophy treated with photodynamic therapy and intravitreal triamcinolone acetonide. PMID- 15805923 TI - Lightning-induced maculopathy. PMID- 15805924 TI - Malignant hypertensive retinopathy studied with optical coherence tomography. PMID- 15805926 TI - Management of posteriorly dislocated posterior chamber intraocular lenses by vitrectomy and pars plana removal. PMID- 15805927 TI - Use of rapid antibiotic susceptibility testing in bacterial endophthalmitis. PMID- 15805930 TI - Commentary on the laws of England: book the third. PMID- 15805931 TI - An overview of medical malpractice litigation and the perceived crisis. AB - In this overview of medical malpractice litigation in the United States, practical and philosophic aspects of the so-called malpractice litigation crisis are addressed. After reviewing the historical, legal rationale for compensating victims of negligent conduct by others, attention is focused on the plight of physicians who are charged with medical negligence and the oppressive insurance premiums that impose a heavy burden on all health care providers, particularly those in the surgical fields. A variety of political solutions advanced to "correct" the problem is reviewed. A historical prospective of malpractice litigation in the United States is presented together with an analysis of various legislative proposals--many of which have already been enacted in various states that will ostensibly "cure" the problems that concern clinicians. Consideration of the various legislative proposals includes: arbitrary limits on pain and suffering awards (caps); elimination of joint and several liability; regulation of attorneys fees; elimination of the collateral source rule; abrogation of punitive damages; proposals for periodic payments; and statutes of repose. Various procedural changes in the processing of malpractice claims are reviewed and analyzed from the perspective of both fairness and efficacy. PMID- 15805932 TI - The defense counsel's perspective. AB - In all likelihood, most orthopedic surgeons today will be the subject of a lawsuit related to the care and treatment of a patient. This article is designed to provide physicians with pertinent material to help prepare for what seems to have become the inevitable--the medical malpractice lawsuit. There are a number of things that can be done to minimize the chance that a surgeon will be sued. Keeping informed about recent developments in orthopaedics, developing a good relationship with patients, and maintaining good documentation are just a few of the ways to help reduce the chances of being named in a potential lawsuit. Additionally, we offer physicians who have been sued assistance in getting through the litigation process, which at times can be daunting. Logical suggestions, such as not discussing a case with anyone other than a spouse and/or an attorney and not altering the medical records and larger concepts, including the importance of developing a relationship with an attorney, understanding the attorney-client privilege, and adequately preparing for the deposition and trial are discussed at length. Although it is not possible to be completely prepared for the day one gets sued, this article should provide insight and information necessary to enable surgeons to face each element of the lawsuit with some knowledge and control. PMID- 15805933 TI - Wrong-site surgery: a preventable complication. AB - All surgical procedures have a risk of complications, many of which cannot be avoided completely regardless of the experience and expertise of the surgeon, the surgical team, or the hospital staff. Wrong-site surgery is a relatively uncommon complication that is easily preventable. The "Sign Your Site" protocol is a simple, straightforward program that requires only a minimal amount of time to eliminate the risk of wrong-site surgery, and it should be standard policy in healthcare institutions. PMID- 15805934 TI - Orthopaedic trauma for the general orthopaedist: avoiding problems and pitfalls in treatment. AB - The Advanced Trauma Life Support program of the American College of Surgeons has developed a de facto standard of care for the first hour of treatment of patients with trauma who are treated in North America. No equivalent for the treatment of orthopaedic trauma has been established. We provide an overview of the important topics every orthopaedist treating patients with musculoskeletal trauma should be familiar with. The topics include: (1) diagnosis of musculoskeletal injury and documentation of physical exam; (2) pelvic ring injuries; (3) open fractures; (4) compartment syndrome; (5) the timing of definitive fixation in the patient with polytrauma; (6) delayed diagnosis of musculoskeletal injury; and (7) deep venous thrombosis prophylaxis. PMID- 15805935 TI - Medicolegal issues in sports medicine. AB - This article reviews medicolegal issues in sports medicine. Nonmusculoskeletal medical conditions discussed include cardiac abnormalities, heat illness, and concussion in the athlete. Musculoskeletal injuries with the greatest potential for litigation, knee dislocation, and cervical spine trauma, are also reviewed. We provide legal case examples and discuss evaluation and treatment strategies to assist the medical team in the care of athletes. PMID- 15805936 TI - Medicolegal issues affecting sports medicine practitioners. AB - The practice of sports medicine represents a unique subspecialty within the discipline of medicine. Practitioners provide care for athletes who engage in activities that may put the athletes at risk for serious injury. Physicians may be held legally liable for not doing a standardized pre-participation evaluation, for not administering adequate on-site or after injury care, or for violating an individual's civil rights by refusing to allow continued participation because of medical risk. The courts now are being asked to define obvious and necessary risks. Medical law addresses the behavior, obligation, and liability-affecting physicians who provide care for athletes. Many ethical and medicolegal issues must be resolved by the legal system because coaches, athletic trainers, on-call physicians, physicians providing pre-participation physicals, and the team physician are being held responsible for injuries sustained during athletic competition. This review is intended to assist all physicians who take care of athletes by outlining their legal obligations and risks. Armed with this knowledge, we expect that the sports medicine physician will be better able to focus on patient care within the context of medicolegal obligations. PMID- 15805937 TI - Medicolegal aspects of the orthopaedic care for shoulder injuries. AB - The medicolegal aspects of orthopaedic care increasingly are becoming an important focus of physicians. Because of the rarity of publications on the subject, I will examine medical litigation in orthopaedics, specifically in regards to shoulder injuries. There are several reasons for recent increases in malpractice litigation. Increased orthopaedic specialization and advanced technologies have raised patient expectations while the media have informed patients of the potential financial rewards of litigation. I will discuss three cases of litigation regarding shoulder injuries including reasons for malpractice and what can be done to avoid it. In Case 1 I examine an implant failure of a rotator cuff repair. The failure was caused by a manufacturing error; therefore, the importance of being familiar with the equipment and the companies that a physician deals with is emphasized in this case. Case 2 is another failed rotator cuff repair; this litigation stemmed from poor patient selection and a lack of preoperative patient-physician communication. Finally, Case 3 involved retained hardware, a surgical mistake that often results in a substantial settlement because it is considered indefensible. These cases highlight the potential for malpractice and can be applied more broadly to all branches of orthopaedics. Most litigation, however, can be avoided with careful diagnostic procedures, greater experience with equipment, and better communication. PMID- 15805938 TI - Complex surgical conditions of the hand: avoiding the pitfalls. AB - Many factors can contribute to a successful result in hand surgery, as is the case in all orthopaedic surgery. An accurate diagnosis with emphasis on a complete history coupled with a detailed physical examination supplemented with the appropriate ancillary studies and proper preoperative surgical planning all are necessary in reaching this goal. An improper diagnosis such as in suspected carpal tunnel syndrome can result in continued symptoms. We will emphasize some of the more complex and difficult conditions in hand surgery that necessitate a high level of clinical acumen. It is hoped that by highlighting these details we may help others avoid common pitfalls and help to achieve an excellent result and avoid litigation. Although these are not the only problems involved in hand surgery, they are complex and can be missed easily. PMID- 15805939 TI - Medicolegal aspects of hip and knee arthroplasty. AB - Hip and knee arthroplasties usually are satisfying for the patient and the surgeon; however, these procedures also have considerable risks for generating a medical malpractice lawsuit. Strict adherence to the standard of care and recently implemented patient safety strategies should reduce the surgeon's liability. Expert technical execution of the surgery, timely evidence-based patient treatment, and detailed documentation in the medical record will not only improve the quality of patient care but also will serve as a strong legal defense should the need arise. PMID- 15805940 TI - Medicolegal aspects of foot and ankle surgery. AB - Medicolegal aspects of orthopaedic foot and ankle surgery, including issues of standard of care, complications, and discrepancies between patient and physician expectations, generally are similar to those encountered in other orthopaedic subspecialties. However, there are some unique aspects involving foot and ankle surgery: the standard of care continues to evolve, some complications are specific to the foot and ankle, and patient expectations of particular foot and ankle procedures may exceed those of the foot and ankle surgeon, who often is confronted with challenging pathologic conditions. This review addresses issues of standard of care and complications as they pertain to current practices of orthopaedic foot and ankle surgery. PMID- 15805941 TI - Pathomechanics in atraumatic shoulder instability: scapular positioning correlates with humeral head centering. AB - The objective was to analyze three-dimensional scapular positioning and glenohumeral centering of normal and atraumatic unstable shoulders. We hypothesized that changes of humeral head position correlate with alterations of scapular positioning. The shoulders of 28 healthy volunteers and 14 patients with atraumatic instability were examined in various arm positions using open magnetic resonance imaging. After segmentation and three-dimensional reconstruction, three dimensional analyses of scapular positioning and humeral head position relative to the glenoid were done. The coefficient of correlation (r) between both parameters was determined using the correlation z test. The glenohumeral to scapulothoracic ratio in the scapular plane was increased in nine of 14 patients and decreased in three patients, whereas the scapular internal rotation in the transverse plane was increased in all unstable shoulders. The unstable shoulders also had malcentering (greater than two times the standard deviation in the healthy volunteers) of the humeral head in the direction of instability during various arm positions. In healthy and unstable shoulders, the correlation between scapular position and glenohumeral positioning was high during passive elevation (r = 0.60-0.87). The high correlation suggests that scapular positioning is relevant for humeral head decentering. Therefore, physiotherapeutic strategy should consider the malpositioning of the scapula and be adapted to the direction of instability. PMID- 15805942 TI - The acromiohumeral and coracohumeral intervals are abnormal in rotator cuff tears with muscular fatty degeneration. AB - We sought to determine how various types of full-thickness rotator cuff tears, fatty degeneration of the rotator cuff muscles, duration of symptoms, and mechanism of injury affect the sizes of the acromiohumeral and coracohumeral intervals. We studied 206 shoulders with rotator cuff tears that had surgical treatment. The acromiohumeral interval (anteroposterior radiograph) and the coracohumeral interval (computed tomogram) were measured on preoperative imaging studies. An abnormal acromiohumeral interval was associated with multiple-tendon rotator cuff tears involving the infraspinatus, fatty degeneration of the supraspinatus or infraspinatus, and duration of symptoms longer than 5 years. An abnormal coracohumeral interval was associated with a combined tear of the supraspinatus and subscapularis and fatty degeneration of the infraspinatus or subscapularis. Fatty degeneration of the infraspinatus therefore was associated with an abnormal acromiohumeral interval and an abnormal coracohumeral interval. Evaluation of a patient who has a diminished acromiohumeral or coracohumeral interval should involve examination with computed tomography or magnetic resonance imaging of the rotator cuff tendons to determine the type of tear and of the rotator cuff muscles to determine the degree of fatty degeneration. LEVEL OF EVIDENCE: Diagnostic study, Level II-1 (development of diagnostic criteria on basis of consecutive patients--with universally applied reference "gold" standard). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805945 TI - Impairment of perineal care functions after long fusions of the lumbar spine. AB - The purpose of this study was to investigate the incidence of perineal care impairment after extended thoracolumbar and thoracolumbosacral spinal fusions. Fourteen adult patients with fusions from the thoracic spine to L5 or the sacrum completed a questionnaire regarding their ability to do perineal care. The mean number of vertebral levels fused was 9.5 (range, 6-16 levels) with five patients having spinal fusion to L5 and nine having fusion to the sacrum. Thirty-six percent (five of 14 patients) reported difficulty doing perineal care after fusion. Maintenance of L5-S1 segmental motion did not seem to reduce occurrence of perineal care problems. We think that extended thoracolumbar fusion and thoracolumbosacral fusion can produce postoperative difficulty in doing perineal care. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series--no, or historical control group). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805948 TI - Curved periacetabular osteotomy for treatment of dysplastic hip. AB - The Bernese periacetabular osteotomy has a considerable rate of postoperative complications such as reflex sympathetic dystrophy, motor nerve palsy, heterotopic ossification, and delayed union of the ilium, which are assumed to be caused by extensive exposure or asphericity of the osteotomy surfaces. To address these issues, we developed the curved periacetabular osteotomy, a modification of the Bernese periacetabular osteotomy which limits dissection, prevents the outside of the ilium from being exposed, and produces osteotomy surfaces with the same curvature. Curved periacetabular osteotomies were done on 128 hips in 118 patients whose average age at the time of surgery was 35.2 years (range, 16-59 years). The average followup was 46 months (range, 24-99 months). The average center-edge angles were 4 degrees (range, -15 degrees -5 degrees ) preoperatively and 35 degrees (20 degrees -55 degrees ) postoperatively, and union of the iliac osteotomy was achieved in all hips. We experienced three asymptomatic pubic nonunions. Dysesthesias occurred in 27 patients along the lateral femoral cutaneous nerve and symptoms resolved in 23 patients within 1 year. The average Harris hip score improved from 72 to 93 points. There were no major complications such as sciatic nerve palsy, abductor dysfunction, or heterotopic ossification. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series--no, or historical control group). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805946 TI - Long-term results of the dial osteotomy in the treatment of high-grade acetabular dysplasia. AB - The dial osteotomy, an acetabular reorientation procedure based on radiographs, was developed by R. H. Eppright for treatment of hip dysplasia; however, long term results are not recorded. The aim of this study was to evaluate retrospectively the results of the dial osteotomy as done in 37 patients (44 hips) at an average followup of 12.6 years. Articular pressures with the application of a time and pressure algorithm were calculated from radiographs to correlate calculated intraarticular pressures with progression of degenerative disease. Clinical results at followup were 32 (73%) satisfactory and 12 (27%) unsatisfactory hips. Six (13%) hips failed between 10-20 years (average 14.7 years). A satisfactory result correlated with the preoperative functional score. Radiographically, the anterior center-edge angle increased from an average of 6.7 degrees to 37.9 degrees. At followup, radiographic indices of degeneration indicated that eight (18%) hips had improved, 18 (41%) had stabilized, and 18 (41%) had deteriorated. Severin indices improved in 21 (48%) hips. Joint space width at followup provided the only correlation between radiographic parameters and clinical result. Contact pressures were reduced from 4.45 MPa to 1.12 MPa. The cumulative exposure to articular pressures averaged 61.6 MPa-years, and did not correlate with radiographic parameters or clinical success. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series--no, or historical control group). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805951 TI - Computer-assisted navigation increases precision of component placement in total knee arthroplasty. AB - In our clinical study, 200 total knee arthroplasties were evaluated to compare the use of the OrthoPilot system with conventional mechanical instrumentation. Long-term outcome of total knee replacement depends mainly on the accuracy of implant positioning and restoration of the mechanical leg axis. Our experience was that navigation could achieve a greater degree of accuracy concerning the aforementioned aspects. Among 513 primary-inserted total knee replacements, 100 navigated knees were compared with 100 conventionally implanted knees after matching the two groups of patients by gender, body mass index, age, preoperative deformities, radiographic findings, and operating time. Three weeks after surgery, the radiographic results were significantly better in the computer assisted group compared with the results in the conventional group when we assessed component positioning in four axes. Only the sagittal tibial component angle was not significantly different. Total knee arthroplasty using the OrthoPilot system led to increased precision of tibial and femoral component positioning in comparison with hand-guided replacement surgery. An additional 10 minutes of operating time was acceptable. Navigation-specific complications were not seen, and the number of outliers decreased. Because computer navigation in orthopaedics is a new technology, data regarding long-term outcomes are not available. LEVEL OF EVIDENCE: Diagnostic study, Level II-1 (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805943 TI - Long-term vertebral changes attributable to postoperative lumbar discitis: a retrospective study of six cases. AB - A clinical, plain radiographic, and magnetic resonance imaging retrospective followup study was done on six patients with postoperative septic L4-L5 lumbar spondylodiscitis to identify the characteristic long-term radiographic findings predicting the clinical outcome. The mean followup was 12 years (range, 9-13 years). Three patients had moderate low-back pain and two were handicapped. One patient had slight peroneal weakness. Flexibility of the lumbar spine was restricted to some extent in all patients. Laminectomy for spinal stenosis (L4 L5) had been done in one patient and reoperation because of recurring disc herniation was done in two patients. Partial or total bone marrow depletion and corresponding enhancement of fat signals in relation to adjacent vertebral bodies, narrowing of the intervertebral disc space, and irregularity or cavitations affecting one or both end plates were long-term findings of magnetic resonance imaging. Plain radiographs and magnetic resonance imaging scans revealed a complete bony fusion in one patient and partial fusion in one patient. In two patients, residual disc protrusion or herniation in relation to the surgically treated disc was evident; three affecting L3-L4 and two affecting L5 S1 disc levels. Septic postoperative lumbar discitis severely hampers clinical recovery after disc operations and results in long-term changes as seen on magnetic resonance imaging scans. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series--no, or historical control group). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805944 TI - Surgical versus pharmacologic treatment of intraspinal gout. AB - A controversy between pharmacologic and surgical treatment of intraspinal gout exists in the literature. If gout is diagnosed timely, pharmacologic therapy may avert the need of surgery. The lack of readily available synovial fluid makes the diagnosis particularly difficult. The purpose of this study was to evaluate the clinical pictures and magnetic resonance imaging features in rapid differentiations of intraspinal gout. I retrospectively evaluated lumbar intraspinal tophaceous gout without the classic radiographic punched-out lesions. Four patients (average age, 65 years) had a history of hyperuricemia with multiple tophaceous deposits in the joints or visceral organs or both. The common presentations were low back pain with or without inflammatory reaction (fever, elevated C-reactive protein level, and mild leukocytosis). The patients also presented with intermittent claudication or radiculopathy of variable duration or both. The gouty tophi yielded homogeneous and hypointense masses on T1- and T2 weighted images, with multiple hypointense speckles. The masses were located in bilateral lumbar facet joints in all patients, with additional midline extension along the ligamentum flavum in three. All patients had uneventful outcomes after surgical decompression and pharmacologic treatment. Rapid deposition of tophi may aggravate nerve compression. If neurologic deficits are found, surgical decompression can provide a satisfactory outcome. LEVEL OF EVIDENCE: Therapeutic study, Level IV. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805947 TI - Intertrochanteric varus osteotomy for osteoarthritis in patients with hip dysplasia: 6 to 28 years followup. AB - There have been few reports regarding midterm to long-term results of intertrochanteric varus osteotomy for treatment of hip dysplasia. We investigated 55 hips in 46 patients, with an average age at surgery of 32 years (range, 12-55 years), for an average followup of 17 years (range, 6-28 years). Preoperative osteoarthritis was Tonnis Grade 0 in seven hips, Grade 1 in 23 hips, Grade 2 in 21 hips, and Grade 3 in four hips. Preoperative anteroposterior radiographs of the 55 hips showed almost perfect congruency between the acetabulum and the femoral head with the hip in an abducted position. Excellent or good clinical results were obtained in 23 hips (42%). Using a Harris hip score less than 70 points or the time until additional surgery as the end point, the cumulative rates of survival were 81% at 10 years, 60% at 20 years, and 50% at 25 years. The clinical results of patients with Grade 0, 1, or 2 osteoarthritis, with good spherical femoral head and mild dysplasia, were acceptable; however, this osteotomy might not be indicated for patients with Grade 3 osteoarthritis or for those with severe acetabular dysplasia. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series--no, or historical control group). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805950 TI - Rotational deformity in varus osteoarthritis of the knee: analysis with computed tomography. AB - Rotational deformities in varus osteoarthritis of the knee were evaluated with computed tomography. Preoperative computed tomography scans of 150 knees (114 patients) having total knee arthroplasty and 31 control knees (20 patients) were included. The femorotibial rotation was quantified using the clinical epicondylar axis of the femur and the tibial tuberosity (patella tendon) as the references. The knees with osteoarthritis were divided into three groups according to the femorotibial angle (Group 1, 0 degrees -9 degrees varus, n = 87; Group 2, 10 degrees -19 degrees varus, n = 51; Group 3, 20 degrees or greater varus, n = 12) and statistically analyzed. Rotational deformities (external rotation of the tibia) existed in knees with osteoarthritis and were larger in knees with increased varus deformities (mean +/- standard deviation, -2.24 degrees +/- 4.19 degrees in Group 1; 0.33 degrees +/- 4.14 degrees in Group 2; and 5.33 degrees +/ 5.71 degrees in Group 3). When the femorotibial rotation of each knee was stratified by the corresponding femorotibial angle, we found a correlation. The information should help minimize the rotational mismatch between the femoral and the tibial components in total knee arthroplasty and elucidate the pathogenesis of varus osteoarthritis of the knee. LEVEL OF EVIDENCE: Diagnostic study, Level III-1 (study of nonconsecutive patients--no consistently applied reference gold standard). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805949 TI - Cemented total knee arthroplasty in patients with juvenile rheumatoid arthritis. AB - The optimal techniques and implants for total knee arthroplasty in patients with juvenile rheumatoid arthritis are controversial. We report the functional outcomes and complications of a series of 17 cemented total knee arthroplasties done by one surgeon during a 10-year period in which off-the-shelf implants were used, the posterior cruciate ligament was excised, and a lateral retinacular release was done. Preoperatively, all knees had severe loss of normal joint space and osteopenia on 3-foot, standing AP radiographs, lateral radiographs, and patellofemoral views. The patients were evaluated after a mean followup of 74 months (range, 36-116 months). The Knee Society scores improved from a mean of 38.9 +/- 23.9 points (range, 10-81 points) preoperatively to 81.9 +/- 16.6 points (range, 39-99 points) postoperatively. Range of motion showed significant improvement in all patients at the most recent followup. Ambulation scores improved significantly; nine of 10 patients (15 knees) were ambulatory after surgery. Complications included two transient regional pain syndromes and one patellofemoral subluxation requiring realignment. Cemented total knee arthroplasty with off-the-shelf implants, excision of the posterior cruciate ligament, and lateral retinacular release in patients with juvenile rheumatoid arthritis can provide substantial improvement in pain, deformity, ambulation, and function. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series--no, or historical control group). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805952 TI - Bilateral total knee arthroplasty increases the propensity to trip on an obstacle. AB - Tripping over an obstacle is the most frequent cause of falls. We examined the effects of total knee arthroplasty on obstacle avoidance success rates in older adults. Obstacle avoidance success rates, body mass index, visual acuity, contrast sensitivity, depth perception, and single-leg stance duration were evaluated in 29 subjects who had bilateral total knee arthroplasties (age range, 72.6 +/- 5.4 years) and 27 age-matched healthy control subjects (age range, 70.6 +/- 5.5 years). The patients who had total knee arthroplasties had a lower obstacle avoidance success rate, lower single-leg stance duration, and greater body mass index than control subjects. Age, contrast sensitivity, and depth perception were not different between patients who had total knee arthroplasties and control subjects. Obstacle avoidance success rates decreased linearly as single-leg stance duration decreased in the control group and across all groups, but not in the group that had total knee arthroplasties. Linear relationships between obstacle avoidance success rates and body mass index existed for all subjects but not for the group that had total knee arthroplasties or the control group individually. Total knee arthroplasty reduces obstacle avoidance success rate, suggesting that persons who have total knee arthroplasties have an increased propensity to trip on an obstacle and fall. Increased body mass index and decreased single-leg stance duration in patients who have total knee arthroplasties are associated with a decreased obstacle avoidance success rate. PMID- 15805955 TI - Treatment of reflex sympathetic dystrophy in children using a prostacyclin analog: preliminary results. AB - The aim of our study was to evaluate Iloprost, a prostacyclin analog, for treatment of reflex sympathetic dystrophy in children not responsive to conservative treatment. Seven female patients with a mean age of 9 years (range, 6-11 years) diagnosed with reflex sympathetic dystrophy Stage II were treated with infusions of Iloprost given on three consecutive days. Additionally, all patients had physiotherapy and were offered psychologic consultation. One day after the final infusion, all seven patients were free of pain and achieved full weightbearing. The side effects of Iloprost were headache in all patients and vomiting in two patients. Two patients experienced relapse--one patient at 3 months and the other patient 5 months after primary treatment--but both were free of pain by 5 days after a second series of infusions. During a mean followup of 30 months all patients remained asymptomatic. Our preliminary results suggest that temporary sympathicolysis with Iloprost, physiotherapy, and psychologic counseling is a safe and an effective treatment of reflex sympathetic dystrophy in children with a long history of symptoms or in those who do not respond to conservative treatment. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series--no, or historical control group). See the Instructions for Authors for a complete description of levels of evidence. PMID- 15805954 TI - The diagnostic accuracy of MRI versus CT imaging for osteoid osteoma in children. AB - Advanced imaging often is obtained in children suspected of having osteoid osteoma. We hypothesized that magnetic resonance imaging gives a falsely aggressive appearance and that computed tomography is better for identifying osteoid osteoma. This is the first prospective blinded study comparatively assessing these imaging studies in children. Twelve preoperative magnetic resonance imaging scans of confirmed cases of osteoid osteoma were collected. Three radiologists blinded to the diagnosis reviewed these images. Only a clinical history of skeletal pain was given. Lesions were classified as benign latent, benign-aggressive, or malignant. Radiologists were asked to list their primary diagnosis (with a confidence level from 1-10). Seven of these 12 children also had computed tomography scans that subsequently were reviewed in similar manner. With computed tomography scans, lesions were accurately identified as benign-latent (15/21 readings, 71%) and as osteoid osteoma (14/21 readings) more frequently than with magnetic resonance imaging scans (7/36 readings, 19%). LEVEL OF EVIDENCE: Diagnostic study, Level III-1 (study of nonconsecutive patients--no consistently applied reference gold standard). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805956 TI - Effect of cyclic pneumatic soft tissue compression on simulated distal radius fractures. AB - We investigated the effect of pneumatic pressure applied to the proximal musculature of the sheep foreleg on load at the site of a transverse osteotomy of the distal radius. The distal radii of 10 fresh sheep foreleg specimens were osteotomized and a pressure sensor was inserted between the two bone fragments. An inflatable cuff, connected to a second pressure sensor, was positioned around the proximal forelimb musculature and the leg then was immobilized in a plaster cast. The inflatable cuff was inflated and deflated repeatedly to various pressures. Measurements of the cuff pressure and corresponding change in pressure at the osteotomy site were recorded. The results indicated that application of pneumatic pressure to the proximal foreleg musculature produced a corresponding increase in load at the osteotomy site. For the cuff pressures tested (109.8 238.4 mm Hg), there was a linear correlation with the load at the osteotomy site with a gradient of 12 mm Hg/N. It is conceivable, based on the results of this study, that a technique could be developed to provide dynamic loading to accelerate fracture healing in the upper limb of humans. PMID- 15805953 TI - Pediatric spinal cord injury without radiographic abnormality: a meta-analysis. AB - Using a meta-analysis, we identified 392 published cases of patients recently diagnosed with spinal cord injuries without radiographic abnormalities (SCIWORA) and studied the epidemiologic, pathophysiologic, clinical, and radiologic data. To describe those at risk for this uncommon syndrome, mainly pediatric patients (90% of the cases) who sustain serious trauma in car accidents, serious falls, sports injuries, or child abuse, we analyzed the reported information in the literature. Magnetic resonance imaging scans may indicate neural (hemorrhages, edema, or both) or extraneural (disc protrusions, extradural hematomas) injuries, and the location and type of the injury. Every patient having magnetic resonance imaging scans had either intraneural or extraneural injury. However, followup magnetic resonance imaging scans are necessary because evidence of the injury might not appear immediately. Late and recurrent spinal cord injuries without radiographic abnormalities are reported. External immobilization for 12 weeks helps patients who are moderately injured and helps prevent recurrence of these types of injuries. Approximately 44% of the patients in our study did not recover whereas complete recovery occurred in 39% of the population. The prognosis can be improved if the syndrome is diagnosed early, so we recommend considering spinal cord injuries without radiographic abnormalities as a possible diagnosis for any child who has a mechanism of injury that suggests trauma to the spine. LEVEL OF EVIDENCE: Therapeutic study, Level III-3 (case-control study). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805958 TI - Fracture callus engulfing a peripheral nerve does not affect its function: an experimental study in rabbits. AB - The fate of a peripheral nerve engulfed in fracture callus is not known. We investigated the impact of envelopment of the sciatic nerve by fracture callus using a New Zealand rabbit femoral fracture model. The sciatic nerve was mobilized and coiled around the ipsilateral femur, which was surgically fractured, shortened, and osteosynthesized. Bony union was achieved, and callus engulfed the sciatic nerve in all animals. Nerve function was evaluated clinically and by conduction studies preoperatively and postoperatively. Although the nerve function in terms of clinical evaluation, amplitude, motor latency, and spontaneous activity deteriorated immediately postoperatively, recovery was evident in the following weeks indicating that the detected nerve dysfunction was attributable to the surgical mobilization. In addition, histologic and quantitative histomorphometric analyses proved that in none of the animals did the callus compress the sciatic nerve whereas an impressive process of axonal regeneration took place despite callus maturation. Results of our study suggest that callus, engulfing a peripheral nerve, does not compress it and the nerve appears to be intact in an osseous canal. This results in preservation of the integrity and function of the nerve, which may have significant clinical applications. PMID- 15805960 TI - The treatment of chronic open osteomyelitis of the tibia in adults. AB - The treatment of 30 consecutive adult patients with chronic draining osteomyelitis of the tibia was reviewed. There were four treatment patterns. Eight patients had local debridement with or without soft tissue coverage procedures or bone graft. Three patients had radical debridement and bone transport using a circular frame. Eight patients were treated by the Papineau grafting technique after debridement. Eleven patients had debridement and circular frame fixation to correct associated nonunion, malunion, or shortening. All patients received long-term antibiotic therapy. At an average of 6 years' followup (minimum, 2 years), two patients had persistent drainage and one patient had an aseptic nonunion. This experience affirms the value of the circular frame, of the Papineau graft, of bone transport, and of long-term antibiotics for treatment of chronic osteomyelitis of the tibia. There was successful limb salvage in all of the patients and successful treatment (fracture healing without drainage) in 27 of 30 patients. LEVEL OF EVIDENCE: Therapeutic study, Level III-1 (case-control study). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805959 TI - Host classification predicts infection after open fracture. AB - We propose that the classification commonly used in patients with osteomyelitis, the Gustilo classification, also is applicable to patients with open fractures as a method of identifying increased risk of infection because of comorbid medical illness. The records of 174 adult patients sustaining open fractures of long bones were retrospectively reviewed. Each patient was sorted into Class A, B, or C based on 14 immune system compromising factors. Class A has no compromising factors, Class B has one or two compromising factors, and Class C has more than three compromising factors. The association between class and infection was examined. The incidence of infection was 4%, 15%, and 30% in patients in Classes A, B, and C, respectively. Patients in Class B were 2.86 times more likely to have an infection, and patients in Class C were 5.72 times more likely to have an infection when both groups of patients were compared with patients in Class A. The Gustilo classification, location of fracture, and tobacco use are all factors associated with infection. Patients in Class B or C are at markedly increased risk of infection and may benefit from additional or alternative therapies that decrease the risk of infection. LEVEL OF EVIDENCE: Prognostic study, Level II-1 (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805963 TI - Indentation stiffness of repair tissue after autologous chondrocyte transplantation. AB - Our main hypothesis was that indentation stiffness of the repair tissue approaches the values of adjacent cartilage 1 year after autologous chondrocyte transplantation. We also wanted to investigate the differences between osteochondritic lesions and full-thickness lesions. Thirty patients with cartilage lesions were operated on with autologous chondrocyte transplantation. The repair was evaluated arthroscopically, indentation stiffness was measured, and clinical evaluations were done. The stiffness of the repair tissue improved to 62% (mean 2.04 +/- 0.83 N, mean +/- SD) of adjacent cartilage (3.58 +/- 1.04 N). Fifty-three percent of the patients graded their knee as excellent or good and 47% of the patients graded their knee as fair at the followup. In six patients the normalized stiffness was at least 80%, suggesting hyaline-like repair. The indentation stiffness of the osteochondritis dissecans lesion repairs (1.45 +/- 0.46 N; n = 7) was less than that of the nonosteochondritis dissecans lesion repair sites (2.37 +/- 0.72 N; n = 19). Gadolinium-enhanced magnetic resonance imaging of the cartilage (dGEMRIC) during followup of four patients suggested proteoglycan replenishment, although all grafts showed low indentation values. Low stiffness values may indicate incomplete maturation or predominantly fibrous repair. The indentation analysis showed that the repair tissue stiffness could, in some cases, reach the same level as the adjacent cartilage, but there was a large variation among the grafts. PMID- 15805957 TI - Tourniquets may increase postoperative swelling and pain after internal fixation of ankle fractures. AB - Tourniquets frequently are used in orthopaedic surgery to provide a bloodless field. There is still controversy among authors regarding the benefits and potential risks of using a tourniquet. The objective of this prospective randomized study was to quantify the effect of tourniquet use on postoperative swelling, pain, and range of motion after open reduction and internal fixation of ankle fractures. Fifty-four patients with closed ankle fractures were treated surgically; 26 patients were operated on using a thigh tourniquet (Group A), and 28 patients had surgery without the use of a tourniquet (Group B). The groups of patients were similar for age, gender, fracture types, and operative procedures. There was no difference in operation time. Using a tourniquet during open reduction and internal fixation of ankle fractures did increase postoperative swelling and postoperative pain at Day 5 and at Week 6 after surgery. We found a trend for a better range of motion in the ankle until the 6-week followup in the nontourniquet group compared with the group in which the tourniquet was inflated. According to these results we do not recommend using a tourniquet for osteosynthesis of ankle fractures. LEVEL OF EVIDENCE: Therapeutic study, Level I 1 (randomized controlled trial). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805961 TI - Improved detection of orthopaedic surgical site infections occurring in outpatients. AB - To test the hypothesis that a simple method of text searching of standard dictated outpatient clinic notes could be used to detect postoperative infections that otherwise were undetected by the traditional hospital-based infection surveillance methods, a retrospective analysis of all clinical notes from the orthopaedic surgery service of a tertiary care medical center without an electronic medical record system was done. Text-searching algorithms were used to screen all outpatient clinic notes for patients seen between September, 2000 and May, 2001. Clinic notes were screened using specific search terms indicative of surgical site infection, to yield a subset of notes that then were manually reviewed. The identified 18,887 notes were reduced to 558 screen-positive notes, of which 347 notes (197 patients) had confirmed surgical site infections on manual review (positive predictive value, 62.2%). Eighty percent of the total joint infections and 54% of the spine infections were not detected through standard hospital-based surveillance. This simple, inexpensive, method for screening clinic notes is effective in improving detection of postdischarge surgical site infections (and, therefore, quality assessment for surgery). It can be implemented in most current clinical settings. LEVEL OF EVIDENCE: Diagnostic study, Level II-1 (development of diagnostic criteria on basis of consecutive patients--with universally applied reference gold standard). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 15805964 TI - Osteogenesis and angiogenesis in regenerating bone during transverse distraction: quantitative evaluation using a canine model. AB - For some patients with a bone defect, curved deformity, or small diameter bone, transverse distraction might be better indicated than bone transport or lengthening. However, detailed quantitative evaluation of osteogenesis or angiogenesis during transverse distraction by creating a well-controlled animal model has not been done. We established a transverse distraction model of the canine tibia. A rectangular cortex was separated for a distraction fragment. Seven days after the operation, 0.5-mm distraction was applied twice daily for 14 days. After a 28-day consolidation period, the animals were sacrificed. Quantitative evaluations of the rates of mineralization and the increases of bone mineral density indicated faster mineralization and earlier corticalization of the regenerating bone in the initial stage of the consolidation period. The average blood vessel volume ratio in the distraction area was more than three times greater than in the intact contralateral tibiae. We hypothesized that adequate preservation of the marrow arteries and stability of the distraction site throughout an experimental period could induce this faster osteogenesis. Our results indicated that the transverse distraction technique was feasible. The transverse distraction technique could be indicated for patients with small diameter bones or with massive bone defects. PMID- 15805962 TI - Intermittent compressive strain may reduce aggrecanase expression in cartilage: a study of chondrocytes in agarose gel. AB - We tested the hypothesis that intermittent compressive strain reduces the catabolic actions of interleukin-1 beta on chondrocyte metabolism at the gene expression level. We investigated the effects of intermittent compressive strain on mRNA expression in bovine chondrocytes cultured in agarose gel supplemented with or without interleukin-1 beta. Fifteen percent compressive strain amplitude was applied to agarose-chondrocyte constructs at a frequency of 1 Hz. In the absence of interleukin-1 beta, the strain caused an increase in the mRNA levels of anabolic factors like aggrecan and Type II collagen, compared with the levels of anabolic factors in unstrained constructs. These results suggest that the strain may stimulate matrix production in normal cartilage. Interleukin-1 beta is a powerful catabolic agent; in unstrained agarose-chondrocyte constructs, interleukin-1 beta caused a decrease in the mRNA levels of anabolic factors. However, interleukin-1 beta also caused an increase in the mRNA of catabolic factors like aggrecanase-1, aggrecanase-2, and matrix metalloproteinase-3. In the presence of interleukin-1 beta, the strain reduced the mRNA levels of aggrecanase 1 and aggrecanase-2. These results suggest that intermittent compressive strain may protect cartilage by suppressing the expression of aggrecanase-1 and aggrecanase-2, which are thought to be the major matrix-degrading enzymes responsible for cleaving aggrecan. PMID- 15805965 TI - Osteoprogenitor cells and osteoblasts are targets for hepatitis C virus. AB - The goal of this study was to determine whether human osteoblasts might harbor the hepatitis C virus. We tested for positive-strand and negative-strand (replicative) hepatitis C virus RNA by reverse transcriptase-polymerase chain reaction, by in situ reverse transcriptase-polymerase chain reaction for intracellular localization of the hepatitis C virus, and by amplicon sequencing in in vitro differentiated mature osteoblasts from STRO-1+ osteoprogenitor cells from patients with chronic hepatitis C and from healthy individuals. We only detected the hepatitis C virus genome in STRO-1+ cells and mature osteoblasts from carriers with chronic hepatitis C, and we found hepatitis C virus negative strands expressed sporadically in these patients. Using in situ hepatitis C virus reverse transcriptase-polymerase chain reaction, we determined that the percentage of infected carrier osteoblasts ranged from 8.0-15.3%. These data provide evidence of hepatitis C virus presence and replication in human osteoprogenitors and osteoblasts, which may have important implications for bone allograft processing. PMID- 15805966 TI - Particles of all sizes provoke inflammatory responses in vivo. AB - The aim of this study was to investigate whether all sizes of wear particles are capable of provoking inflammatory responses and whether there are different responses among different particle sizes. The knees of 40 female Balb/c mice were injected with polystyrene particles of three different diameters, 0.5 microm, 2.0 microm, and 75 microm, using a 0.1% vol/vol concentration. Seven days after particle injection, assessment of the synovial microcirculation using intravital microscopy, and histologic examination, were done. All the mice injected with polystyrene particles had enhanced leukocyte-endothelial cell interactions and histologic scores regardless of particle size when compared with control animals injected with sterile phosphate buffered saline. Polystyrene particles 0.5 microm in size provoked stronger membrane thickening and increased leukocyte-endothelial cell interactions than 75-microm particles. The fraction of rolling leukocytes was enhanced in the 2.0-microm particle group when compared with the 75-microm particle group. These results indicate that polystyrene particles of all sizes (0.5 microm, 2.0 microm, and 75 microm) are capable of inducing an inflammatory response. Small particles (0.5 microm, 2.0 microm) seem to provoke a stronger inflammatory response than larger particles (75 microm) in conditions with equal particle volume. PMID- 15805967 TI - A hand mass that became painful 13 years after onset. PMID- 15805968 TI - Enlarging knee mass in a 13-year-old girl. PMID- 15805969 TI - Right knee pain in a 13-year-old boy. PMID- 15805976 TI - Spinal stenosis. PMID- 15805984 TI - Advances in upper airway diseases and allergen immunotherapy. AB - Evidence of an increased prevalence of rhinitis in patients with asthma, and asthma in patients with rhinitis, supports the 1 airway concept. However, there are basic differences between the upper and lower airways, such as the virtual absence of remodeling in the nose compared with the bronchi, despite the presence of similar inflammation. Etiologic factors in chronic rhinosinusitis (CRS) attract increasing interest. Peripheral blood monocytes from patients with CRS release IL-4, IL-13, and IFN-gamma on stimulation with fungal antigens, especially those from Alternaria. This is not seen with cells from normal controls. However, a double-blind trial of intranasal amphotericin in CRS was negative. Evidence continues to accumulate of the pivotal role of regulatory T lymphocytes secretion of IL-10 in the response to allergen immunotherapy. In patients with asthma and house dust mite allergy who are receiving appropriate pharmacotherapy and have instituted environmental controls, house dust mite immunotherapy provides marginal additional benefits in asthma control. Immunotherapy with cat dander extract at a maintenance dose containing 15 microg Fel d 1 produces a more consistent immunologic response than with maintenance doses containing 3.0 microg, whereas doses containing only 0.6 microg are no more effective than placebo. Sublingual immunotherapy for seasonal grass allergy can be safely administered by general practitioners, but symptom relief begins only in the second season of therapy. Sublingual immunotherapy for seasonal grass allergy in children reduced symptoms and onset of new asthma symptoms but, again, beginning only in the second year of treatment. A course of 6 weekly injections of ragweed Amb a 1 bound to cytosine phosphorothionate guanosine containing DNA produced a shift from T H 2 to T H 1 cytokine release both in peripheral blood cells and in the nose after allergen challenge. No symptom improvement was seen the first year, but symptoms were reduced the second year without further treatment. PMID- 15805977 TI - Facet-mediated pain. PMID- 15805980 TI - Assessing respiratory function in young children: Developmental considerations. AB - The purpose of this review is to provide practitioners and clinical investigators with an update on methods of assessing respiratory function in young children. The importance of this topic is presented in light of the natural history of asthma, as well as maturational changes that occur early in life in terms of airway development. Models of disease are cited to support the concept that injury of the mammalian airway early in postnatal life might have far-reaching consequences in terms of control of airway caliber and responsiveness. The methods currently available to measure respiratory function in our younger patients are outlined. The ability of children to perform the maneuvers necessary for this testing is considered as a function of age. Areas in which research and development are needed are highlighted. PMID- 15805978 TI - Minimally invasive spine surgery for low back pain. PMID- 15805982 TI - Effects of viral respiratory infections on lung development and childhood asthma. AB - Viral infections are closely linked to wheezing in infancy, and those children with recurrent virus-induced wheezing episodes are at great risk for chronic childhood asthma. Infancy is a time of increased susceptibility to viral infections, and this stage is also characterized by pulmonary alveolar multiplication and extensive remodeling of the airways to accommodate growth. This coincidence, together with the observation that children with asthma can have structural lung changes and functional deficits at an early age, suggests that viral infections could adversely affect lung development. Inflammatory mediators induced by viral infection are known to have effects on the remodeling process, suggesting a plausible mechanism to support this theory. Furthermore, animal models of viral infection during lung growth and development suggest that developmental factors are important in determining the consequences of infection on long-term lung function. Greater understanding of the effects of viral infections on lung development and growth in early childhood might lead to the discovery of additional strategies for the prevention of recurrent wheezing and chronic asthma. PMID- 15805986 TI - The role of air pollution in asthma and other pediatric morbidities. AB - A growing body of research supports the role of outdoor air pollutants in acutely aggravating chronic diseases in children, and suggests that the pollutants may have a role in the development of these diseases. This article reviews the biologic basis of children's unique vulnerability to highly prevalent outdoor air pollutants, with a special focus on ozone, respirable particulate matter (PM 2.5 [<2.5 microm in diameter] and PM 10 [<10 microm in diameter]), lead, sulfur dioxide, carbon monoxide, and nitrogen oxides. We also summarize understanding regarding health effects and molecular mechanisms of action. Practitioners can significantly reduce morbidity in children and other vulnerable populations by advising families to minimize pollutant exposures to children with asthma, or at a broader level by educating policymakers about the need to act to reduce pollutant emissions. Management of children with asthma must expand beyond preventing exposures to agents that directly cause allergic reactions (and therefore can be diagnosed by means of skin tests) and must focus more attention on agents that cause a broad spectrum of nonspecific, generalized inflammation, such as air pollution. PMID- 15805985 TI - Facing the challenges of childhood asthma: what changes are necessary? PMID- 15805987 TI - Progression of asthma in childhood. AB - In general, asthma runs a variable course. However, there are children with asthma who are at risk for progressive disease that can lead to failure to attain peak lung growth and eventually can result in clinical and functional impairment. We present a case of a 14-year-old girl who has demonstrated a deterioration in lung function at least in the past 6 years. This has been associated with increasing medication requirements, including chronic oral corticosteroid and a steroid-sparing medication, the presence of comorbidities, and the occurrence of frequent exacerbations. The patient has evidence of airway inflammation and remodeling based on biomarkers and biopsy findings. It remains a challenge to determine patients at risk for irreversible airflow obstruction, presumably a result of airway remodeling. Histopathologic studies from biopsy samples provide the best evidence for this. However, bronchoscopy and biopsy require technical expertise and cannot be routinely used. Clinically, once a patient increases medication requirements, this should serve as a marker of disease progression. Biomarkers of airway inflammation such as exhaled nitric oxide, exhaled breath condensates, and induced sputum analysis may have the potential to monitor disease progression. Additional efforts are necessary to understand the mechanisms involved in this phenomenon and to discover specific modalities of treatment to target the structural changes of airway remodeling. PMID- 15805988 TI - Understanding mild persistent asthma in children: the next frontier. AB - Limitations in asthma prevalence studies and difficulties in diagnosing pediatric asthma lead to uncertainty over the full extent of mild persistent asthma in children and adolescents. Although recent surveys have reported that the majority of pediatric patients with asthma in the United States and Europe have symptoms consistent with mild disease, these surveys have limitations in design. Thus, the true prevalence of mild asthma remains unknown. It is unclear whether children with mild persistent asthma progress to more severe asthma, but the risk of severe asthma exacerbations seems to be unrelated to the symptom severity. Clinical studies restricted to pediatric patients with mild asthma are limited, but available data do suggest substantial morbidity of mild persistent asthma in this population and support inhaled corticosteroid intervention. There is a need for further investigation into the true prevalence of mild persistent asthma in children and adolescents, and optimal treatment. PMID- 15805989 TI - Bias in observational study of the effectiveness of nasal corticosteroids in asthma. AB - BACKGROUND: A recent observational study suggests that intranasal corticosteroids used to treat allergic rhinitis are effective at preventing asthma outcomes, such as emergency visits. The approach to data analysis may have led to biased results because of misclassification of immortal time. OBJECTIVE: To illustrate the bias in the cohort approach and to present the proper time-dependent analysis by replicating the recent study using data from another source. METHODS: From an existing cohort of 30,569 patients with asthma age 5 to 44 years and identified from the Saskatchewan Health databases (1975-1997), we formed the cohort of all subjects who were in the source population between January 1, 1989, and December 31, 1991. Subjects were followed to the first asthma hospitalization. All prescriptions dispensed during follow-up were identified. We replicated the time fixed approach to data analysis used in the recent study and compared it with time-dependent approaches. RESULTS: The cohort included 20,173 subjects, of whom 1849 were hospitalized for asthma between January 1, 1989, and December 31, 1991. The time-fixed approach misclassified more than 5000 person-years of follow-up, corresponding to 44% of the exposed person-time. As a result, the rate ratio of asthma hospitalization after any use of nasal corticosteroids (NCSs) was 0.57 by the biased time-fixed approach compared with 1.13 by the proper time-dependent approach. The time-fixed approach produced a paradoxical protective effect of NCS with 1 or less canisters dispensed per year (odds ratio, 0.47), which was further exaggerated when the cohort was extended to 5 years (odds ratio, 0.33). Adjusted time-dependent analyses found no protective effect, even when NCSs were dispensed regularly (rate ratio, 1.10; 95% CI, 0.54-2.21). CONCLUSION: The time-fixed approach to the analysis of the effectiveness of NCSs on asthma outcomes leads, by its inherent misclassification of immortal time, to a considerable exaggeration of the protective effect of these medications in preventing severe asthma exacerbations. PMID- 15805992 TI - The relationship between vaccine refusal and self-report of atopic disease in children. AB - BACKGROUND: In the last 3 decades, there has been an unexplained increase in the prevalence of asthma and hay fever. OBJECTIVE: We sought to determine whether there is an association between childhood vaccination and atopic diseases, and we assessed the self-reported prevalence of atopic diseases in a population that included a large number of families not vaccinating their children. METHODS: Surveys were mailed to 2964 member households of the National Vaccine Information Center, which represents people concerned about vaccine safety, to ascertain vaccination and atopic disease status. RESULTS: The data included 515 never vaccinated, 423 partially vaccinated, and 239 completely vaccinated children. In multiple regression analyses there were significant ( P < .0005) and dose dependent negative relationships between vaccination refusal and self-reported asthma or hay fever only in children with no family history of the condition and, for asthma, in children with no exposure to antibiotics during infancy. Vaccination refusal was also significantly ( P < .005) and negatively associated with self-reported eczema and current wheeze. A sensitivity analysis indicated that substantial biases would be required to overturn the observed associations. CONCLUSION: Parents who refuse vaccinations reported less asthma and allergies in their unvaccinated children. Although this relationship was independent of measured confounders, it could be due to differences in other unmeasured lifestyle factors or systematic bias. Further research is needed to verify these results and investigate which exposures are driving the associations between vaccination refusal and allergic disease. The known benefits of vaccination currently outweigh the unproved risk of allergic disease. PMID- 15805991 TI - Pulmonary chemokines and their receptors differentiate children with asthma and chronic cough. AB - BACKGROUND: Cough is a frequent symptom in children, but the differentiation of asthmatic cough from cough of other origins can be difficult. Chemokines recruit T lymphocytes to inflamed tissues, and the corresponding chemokine receptors are differentially expressed on T H 1 and T H 2 cells. OBJECTIVE: We sought to determine whether levels of T H 1/T H 2-related chemokines and their receptors differ in bronchoalveolar lavage fluid (BALF) from 12 children with allergic asthma, 15 nonatopic children with chronic cough, and 10 children without airway disease. METHODS: The T H 1-related (IFN-gamma-inducible protein of 10 kd [IP 10], IFN-gamma-inducible T cell alpha chemoattractant [ITAC], monokine induced by IFN-gamma [Mig], and IFN-gamma) and T H 2-related (thymus- and activation regulated chemokine [TARC], macrophage-derived chemokine [MDC], IL-5, and IL-4) chemokines and cytokines were quantified in BALF by ELISA and a particle-based multiplex array. Percentages of pulmonary lymphocytes expressing CXCR3 + and CCR5 + (T H 1) and CCR4 + and CCR3 + (T H 2) chemokine receptors were determined in BALF by flow cytometry. RESULTS: Pulmonary CCR4 + CD4 + cells and levels of TARC and MDC were significantly increased in asthmatic children versus children with chronic cough or without airway disease. In asthmatic children CCR4 + CD4 + cells correlated positively with levels of TARC, MDC, and serum IgE levels and negatively with FEV 1 . In contrast, CXCR3 + CD8 + cells and levels of ITAC were significantly increased in children with non-atopic chronic cough compared with the other groups. In children with chronic cough, CXCR3 + CD8 + cells correlated with levels of ITAC and IFN-gamma. CONCLUSION: Pulmonary CCR4 + CD4 + and CXCR3 + CD8 + cells and their ligands TARC, MDC, and ITAC clearly differentiate asthmatic children from nonatopic children with chronic cough. The analysis of these markers could facilitate the diagnostic discrimination of asthma versus other reasons for chronic cough in children. PMID- 15805993 TI - Protection by budesonide and fluticasone on allergen-induced airway responses after discontinuation of therapy. AB - BACKGROUND: Treatment with inhaled steroids is an effective method of reducing bronchoconstriction and airway inflammation after allergen challenge. However, the duration of the protective effects of inhaled steroids after discontinuation of therapy has not been established. OBJECTIVE: We sought to evaluate the protective effect of 1 week of inhaled steroid therapy against inhaled allergen challenge 12 hours after discontinuation of therapy. METHODS: In this randomized, double-blind, placebo-controlled crossover trial, 26 asthmatic subjects (>18 years old) not using inhaled steroids were administered 200 microg of budesonide twice daily, 200 microg of fluticasone twice daily, or placebo twice daily for 1 week. Twelve hours after discontinuation of therapy, subjects were administered an inhaled allergen challenge. Each treatment period was separated by a 3-week washout period. RESULTS: When compared with placebo (26% +/- 14%), there was a slight but significant protection against the allergen-induced early response after fluticasone treatment (19% +/- 10%, P = .001) but not after budesonide treatment (23% +/- 13%, P = .08). However, when the area under the curve for the early airway response was examined, there was no difference between the 2 drugs in the amount of protection ( P = .62). Partial protection was demonstrated against the late-response allergen-induced sputum eosinophilia with both treatments ( P = .001). By contrast, no protection was observed against allergen induced airway hyperresponsiveness for either treatment. CONCLUSIONS: The protective effects of inhaled steroids against allergen-induced early responses, airway eosinophilia, and allergen-induced airway hyperresponsiveness are partially or completely lost as early as 12 hours after discontinuation of therapy. PMID- 15805990 TI - Sputum eosinophil counts predict asthma control after discontinuation of inhaled corticosteroids. AB - BACKGROUND: Although inhaled corticosteroids (ICSs) are effective in preventing deterioration in asthma control, at least half of subjects with mild-to-moderate asthma will remain stable when these agents are discontinued. OBJECTIVE: We sought to determine whether noninvasive markers of inflammation predict which individuals maintain asthma control after discontinuation of ICSs. METHODS: We analyzed data obtained from 164 subjects with mild-to-moderate asthma who participated in a 16-week trial comparing the effects of continued ICS use with the effects of a switch to salmeterol or placebo. RESULTS: In comparison with continued ICS use, a switch to salmeterol or placebo was associated with increased rates of asthma deterioration over 16 weeks (9.3% vs 24.1% and 37.5%, respectively; P = .04 and P < .001, respectively). We found that neither exhaled nitric oxide nor methacholine PC 20 , when measured at randomization or 2 weeks after randomization, were significant predictors of subsequent asthma control in subjects who discontinued ICSs. However, both induced sputum eosinophil counts measured 2 weeks after a switch from ICS to placebo and changes in sputum eosinophil counts from before cessation of ICSs to after a switch to placebo predicted subsequent asthma deterioration (area under the receiver-operating characteristic curve, 0.771 [ P < .001] and 0.825 [ P < .001], respectively). CONCLUSION: On the basis of a model treatment strategy, we estimate that allocating subjects to ICS therapy on the basis of changes in sputum eosinophil counts after a trial discontinuation could allow 48% of subjects with mild-to moderate asthma to discontinue ICS therapy without an increased risk of asthma deterioration over a period of at least 14 weeks. PMID- 15805994 TI - Variation in total and specific IgE: effects of ethnicity and socioeconomic status. AB - BACKGROUND: Asthma is common in minority and disadvantaged populations, whereas atopic disorders other than asthma appear to be less prevalent. It is unclear whether the same holds true for objective markers of sensitization. OBJECTIVE: To determine the association of asthma, atopic disorders, and specific sensitization with race and socioeconomic factors. METHODS: We analyzed total and specific IgE among 882 women (577 white, 169 black, and 136 Hispanic) who delivered a child at a large tertiary hospital in Boston, Mass, and who were screened for participation in a family and birth cohort study. Race/ethnicity and other characteristics were obtained from screening questionnaires. Addresses were geocoded, and 3 census-based geographic area socioeconomic variables were derived from block group information from the 1990 US Census. RESULTS: Black and Hispanic women were more likely to come from areas with low socioeconomic indicators and were more likely to have asthma than white women. However, these women were less likely to have hay fever and eczema than their white counterparts. Compared with white women, black women had higher mean total IgE levels; had greater proportions of sensitization to indoor, outdoor, and fungal allergens; and were more than twice as likely to be sensitized to > or =3 aeroallergens. CONCLUSION: The racial/ethnic disparities in atopic disorders may represent either underdiagnosis or underreporting and suggest that allergy testing may be underused in some populations. Differences in total IgE levels and specific allergen sensitization are likely a result of the complex interplay between exposures associated with socioeconomic disadvantage. PMID- 15805996 TI - Exhaled breath condensate cysteinyl leukotrienes are increased in children with exercise-induced bronchoconstriction. AB - BACKGROUND: It is recognized that airway inflammation has a central role in the pathogenesis of asthma, but how it relates to exercise-induced bronchoconstriction (EIB) is not completely understood. OBJECTIVE: The aim of our study was to investigate the relationship between EIB and baseline concentrations of cysteinyl leukotrienes (Cys-LTs) and other inflammatory markers in exhaled breath condensate (EBC). METHODS: EBC was collected, and the fraction of exhaled nitric oxide (FE NO ) was measured in a group of 19 asthmatic children, after which they performed a treadmill exercise test. Fourteen healthy children were enrolled as control subjects. RESULTS: The asthmatic children were divided into the EIB group (decrease in FEV 1 , > or =12%) and the non-EIB group. The EBC was analyzed for the presence of Cys-LTs, leukotriene B 4 , and ammonia. Asthmatic patients with EIB (mean FEV 1 decrease, 23% +/- 3%) had higher Cys-LT concentrations than either asthmatic patients without EIB or control subjects (42.2 pg/mL [median] vs 11.7 pg/mL and 5.8 pg/mL; P < .05 and P < .001, respectively). Ammonia concentrations were lower in both the EIB and non-EIB groups than in control subjects (253.2 microM and 334.6 microM vs 798.4 microM; P < .01 and P < .05, respectively). No difference in EBC leukotriene B 4 levels was found among the 3 groups. Both asthmatic groups had higher FE NO levels than control subjects ( P < .001). EBC Cys-LT ( P < .01; r = 0.7) and FE NO ( P < .05; r = 0.5) values both correlated significantly with the postexercise FEV 1 decrease. CONCLUSION: this study shows that EBC Cys-LT values are higher in asthmatic children with EIB and correlate with the decrease in FEV 1 after exercise. These findings suggest that the pathways of both Cys-LT and nitric oxide are involved in the pathogenesis of EIB. PMID- 15805995 TI - IL-5 and thromboxane A2 receptor gene polymorphisms are associated with decreased pulmonary function in Korean children with atopic asthma. AB - BACKGROUND: Asthmatic airways undergo chronic inflammatory cell infiltration by T cells and eosinophils, which results in sustained airway hyperresponsiveness. IL 5 is important for eosinophil-induced airway inflammation and airway hyperresponsiveness. Thromboxane A2 and its receptor, TBXA2R, are involved in constriction of respiratory smooth muscles and may play a role in thickening and remodeling of airways, which contributes to the severity of asthma. The relationship between IL-5 and TBXA2R gene polymorphisms and pulmonary function in children with asthma has rarely been examined. OBJECTIVE: To determine whether IL 5 (T-746C) and TBXA2R (T924C) gene polymorphisms are associated with asthma phenotype and pulmonary function in Korean children with atopic and nonatopic asthma. METHODS: We conducted an association study between known polymorphisms of IL-5 (T-746C) and TBXA2R (T924C) and asthma phenotype and the parameters of atopy and pulmonary function in atopic and nonatopic Korean children with asthma. The subjects were 240 atopic children with asthma, 70 nonatopic children with asthma, and 106 nonatopic healthy children. Asthma phenotypes and bronchial responsiveness to methacholine were determined by a physician. IL-5 and TBXA2R gene polymorphisms were determined by genotyping by using PCR-RFLP assays. RESULTS: The genotype frequencies of IL-5 and TBXA2R polymorphisms did not differ between healthy controls and atopic or nonatopic children with asthma. A significant association was observed between the IL-5 polymorphism and forced expiratory flow at 25% to 75% of forced vital capacity (FEF 25-75% ; %; P = .002), and between the TBXA2R polymorphism and FEV 1 (%; P = .035) and FEF 25-75% (%; P = .042) in children with atopic asthma, whereas no such association between the polymorphisms and lung function was observed in nonatopic or control children. In atopic children with asthma, we identified a significant gene-gene interaction in that the combination of the IL-5 (T-746C) and TBXA2R (T924C) mutant alleles was shown to be associated with reduced pulmonary function as determined by FEF 25-75% (%) measurement. CONCLUSION: The current study indicates that IL-5 (T-746C) and TBXA2R (T924C) polymorphisms alone are associated with spirometric markers of asthma severity, whereas they are not associated with presence of asthma per se. In addition, the data suggest that an interaction between IL-5 and TBXA2R genes may contribute to the severity of asthma, especially atopic asthma. These results suggest that IL-5 and TBXA2R genes may be disease-modifying genes in Korean children with atopic asthma. PMID- 15805998 TI - Eotaxin-2 and eotaxin-3 expression is associated with persistent eosinophilic bronchial inflammation in patients with asthma after allergen challenge. AB - BACKGROUND: Eotaxin-1, eotaxin-2, and eotaxin-3 are chemokines involved in the activation and recruitment of eosinophils through activation of their main receptor, CC chemokine receptor 3. The differential roles of these chemokines still remain to be established. It has been suggested that eotaxin-1 is an important mediator in the early phase of allergen-induced recruitment of eosinophils into the airways. Eotaxin-2 and eotaxin-3 might play a role in the subsequent persistence of allergen-induced bronchial eosinophilia. OBJECTIVE: The aim of this study was to determine the expression of eotaxins and eosinophil counts in the bronchial mucosa of subjects with mild asthma after resolution of the late-phase asthmatic response (LAR). METHODS: The expression of eotaxins and eosinophil counts were determined in bronchial biopsy specimens obtained from 10 subjects with mild asthma 48 hours after diluent and allergen challenge by using immunohistochemistry. Positively stained cells were counted in a 125-mum-deep zone of the lamina propria. RESULTS: Eotaxin-2 and eotaxin-3 expression in bronchial mucosa was significantly increased 48 hours after allergen challenge ( P = .001 and P = .013, respectively). At this time point, when marked tissue eosinophilia was still present, these increases were positively correlated with the magnitude of the LAR ( r = 0.72, P = .019 and r = 0.64, P = .046, respectively). Furthermore, eotaxin-2 expression was associated with the number of eosinophils after allergen challenge ( r = 0.72, P = .018). CONCLUSION: Our findings suggest that eotaxin-2 and eotaxin-3 might account for the persistence of bronchial eosinophilia after resolution of the LAR. PMID- 15805999 TI - Out-of-season recollection of drug use for seasonal IgE-mediated rhinitis: useful but an overestimation. AB - BACKGROUND: The history of seasonal, IgE-mediated (allergic) rhinitis is often obtained postseasonally as a retrospective assessment. It is not known whether recollected drug use is representative for the in-seasonal medication history. OBJECTIVES: To investigate the agreement between in-seasonal and out-seasonal reports on drugs and drug doses, and to investigate whether retrospective assessment was sufficiently sensitive to detect the effect of intervention. MATERIAL AND METHODS: Thirty-five patients allergic to birch pollen were allocated to allergen-specific immunotherapy (SIT) or placebo in a double-blind study. Drug use was recorded daily during the season 2000 and out-seasonally 6 months after the seasons 1999 and 2000. The mean daily drug use was transformed into a 4-point categorical scale for simplicity and was calculated for the out seasonal rating and for 6 in-seasonal periods of different durations. In-seasonal and postseasonal ratings were compared with the Cohen weighted kappa (kappa w). RESULTS: We observed a high level of agreement between in-seasonal and retrospective reports of drugs used (kappa w > 0.60) and drug doses (kappa w > 0.50). The highest agreement was observed for the in-seasonal day, week, or 2 weeks with the highest drug intake. Out-seasonal ratings significantly overestimated the daily oral and nasal antihistamine use compared with in seasonal periods >2 weeks. Despite being comparable pretreatment, only the SIT group had a significant decrease in recollected total drug use during SIT. CONCLUSION: Out-seasonal recollection of hay fever drugs represented a period of a maximum 14 days. It appeared sufficiently sensitive to detect the effect of intervention. PMID- 15806000 TI - An intranasal Syk-kinase inhibitor (R112) improves the symptoms of seasonal allergic rhinitis in a park environment. AB - BACKGROUND: R112 inhibits Syk kinase, a transducer of signaling through the Fcepsilon receptor of mast cells, blocking mast cell responses to allergic stimuli. OBJECTIVE: Examine the efficacy and safety of intranasal R112 in volunteers with symptomatic seasonal allergic rhinitis compared with a placebo in a park setting. METHODS: In this double-blind, placebo-controlled study of 319 volunteers with seasonal allergic rhinitis, 160 were randomized to intranasal R112 and 159 to a vehicle control during 2 days at 2 separate locations in spring 2004. Subjects were evaluated for symptoms of allergic rhinitis (i.e., sneezes, runny nose/sniffles, itchy nose, stuffy nose) on the basis of a possible maximum score of 32 for the Global Symptom Complex (GSC) scale. The primary outcome evaluated was the difference in the reduction in GSC (area under the curve over a period of 8 hours) from baseline between R112 and vehicle placebo. RESULTS: At baseline, the combined GSC was approximately 18/32 and equal between treatment groups. After 8 hours (dosing 3 mg/nostril every 4 hours x 2), R112 significantly reduced the GSC compared with placebo (7 vs 5.4 units, respectively; P = .0005). Each individual symptom combined to form the GSC was also significantly improved in the R112 group compared with control ( P < .05). As early as 45 minutes after dosing, R112 showed a significant improvement in symptoms over placebo, and the duration of action exceeded 4 hours. Adverse effects were indistinguishable between the groups and clinically insignificant. CONCLUSION: Intranasal R112 was effective in this park study and is a promising new treatment for seasonal allergic rhinitis. PMID- 15806001 TI - Endotoxin exposure and atopic sensitization in adult pig farmers. AB - BACKGROUND: Recent studies have reported a low prevalence of atopic sensitization and respiratory allergy in children growing up on farms. OBJECTIVES: We sought to evaluate the dose-response relationship between endotoxin and atopic sensitization in adult farmers and to assess the effect on respiratory health outcomes. METHODS: Data on endotoxin exposure and serum IgE levels were available for 162 pig farmers from a cross-sectional case-control study, with case selection on the basis of respiratory symptoms. Exposure to endotoxin was modeled in detail, and respiratory health effects were assessed during a medical examination. Exploratory analysis was done by using nonparametric modeling and was followed by classical parametric regression. RESULTS: IgE to one or more common allergens was detected in sera from 28 (17%) farmers. The average (geometric mean) total serum IgE levels was 37 IU/mL (geometric SD, 4 IU/mL). A strong inverse relationship was found between endotoxin and sensitization to common allergens for exposures of 75 ng/m 3 or less, with an odds ratio of 0.03 (95% CI, 0.0-0.34) for a 2-fold increase in endotoxin. For endotoxin exposure of greater than 75 ng/m 3 , the association was weak (odds ratio, 1.2 [95% CI, 0.38 3.6]). No association was found between endotoxin exposure and total serum IgE levels. Endotoxin was associated with increased airway hyperresponsiveness to histamine and lower lung function in sensitized farmers, without evidence of a nonlinear relationship. CONCLUSIONS: The prevalence of atopic sensitization in adult pig farmers is low. Endotoxin or related exposures might protect from sensitization, even in an adult working population exposed to high levels of endotoxin, but is a risk factor for increased airway hyperresponsiveness and low lung function. PMID- 15805997 TI - Asthmatic bronchial epithelium activated by the proteolytic allergen Der p 1 increases selective dendritic cell recruitment. AB - BACKGROUND: Airway dendritic cells (DCs) are crucial for allergen-induced sensitization and inflammation in allergic asthma. After allergen challenge, an increased number of DCs is observed in airway epithelium from patients with allergy. OBJECTIVE: Because Der p 1, a cysteine protease allergen from Dermatophagoides pteronyssinus , induces chemokine production by bronchial epithelial cells (BECs), the purpose of this investigation was to evaluate the capacity of BEC exposed to Der p 1 to recruit DCs. METHODS: Chemotactic activity of BEAS-2B, a bronchial epithelial cell line, and BECs from nonatopic controls and patients with allergic asthma was evaluated on the migration of precursors, immature and mature monocyte-derived DCs (MDDCs), and CD34 + -derived Langerhans cells (LCs). RESULTS: C-C chemokine ligand (CCL)-2, CCL5, and C-X-C chemokine ligand 10 production by BEAS-2B and BEC was increased after Der p 1 exposure, whereas the proenzyme proDer p 1 devoid of enzymatic activity had no effect. Der p 1 stimulation of BEAS-2B and BEC from both groups increased significantly the recruitment of MDDC precursors, depending on CCL2, CCL5, and C-X-C chemokine ligand 10 production. In a reconstituted polarized epithelium, apical application of Der p 1 enhanced MDDC precursor migration into the epithelial layer. Moreover, Der p 1 stimulation of BEC from patients with asthma but not from controls increased the migration of LC precursors, mainly dependent on CCL20 secretion. No migration of immature and mature DCs was observed. CONCLUSION: These data confirmed that BECs participate in the homeostasis of the DC network present within the bronchial epithelium through the secretion of chemokines. In allergic asthma, upregulation of CCL20 production induced LC recruitment, the role of which remains to be determined. PMID- 15806007 TI - Age-related changes in the cellular composition of the thymus in children. AB - BACKGROUND: T-cell development in the thymus is an extensively studied subject, mainly in mice. Nevertheless, the normal composition and cell numbers of the noninvoluted human thymus are largely unknown. OBJECTIVE: We aimed to gain insight into age-related changes in different thymic subpopulations and to provide reference values for the distribution of thymocyte subsets. The composition of the normal thymus may serve as a reference for thymi in pathological conditions and may aid diagnoses of immunodeficiency diseases. METHODS: Thymic lobes of 70 children (58 immunologically normal and 12 diseased), ranging in age from 8 days to 8 years old, were studied by 4-color flow cytometric analysis. Detailed staining and gating strategies allowed us to dissect small subsets, including immature CD4(-) CD8(-) populations and thymic B, natural killer, and T-cell receptor gammadelta + cells. RESULTS: We demonstrate that distribution of thymocyte subsets changes with age and correlates with age related fluctuations of T-lymphocyte counts in peripheral blood. Thymi of children 3 to 6 months old appear to be the most active: they have high numbers of total thymocytes, the highest percentage of double-positive cells, and large numbers of CD34 + progenitors in their thymi. Furthermore, we show that the human thymus is a site for B-cell development, because all B-cell progenitor stages that can be found in the bone marrow are also present in the thymus. CONCLUSION: We conclude that T-cell development in children is a dynamic process, answering the demands of a maturing and expanding immune system. PMID- 15806003 TI - Influences of earlier adherence and symptoms on current symptoms: a marginal structural models analysis. AB - BACKGROUND: The morbidity and mortality associated with asthma are suspected to be a result, in part, of poor adherence to inhaled corticosteroid regimens. One influence on adherence may be the perception of symptoms. Because symptoms and adherence affect each other over time, a conventional statistical approach for studying these relationships may provide biased results. OBJECTIVE: To understand the influence of previous asthma symptoms and previous adherence on current symptoms. METHODS: A total of 76 adults, mean age 48 years +/- 15 years, with moderate or severe persistent asthma underwent 6 weeks of electronic monitoring of their use of inhaled corticosteroids and completed a daily symptom diary. We estimated the effect of earlier adherence on final symptoms by using marginal structural models, estimated by using a weighted estimation technique. RESULTS: Morning was better than evening adherence, which declined over the observation period. The variability of adherence appeared to increase over the observation period. In addition, earlier adherence predicted current adherence more strongly than earlier symptoms predicted current adherence. There was no overall significant relationship between cumulative adherence and final symptoms. CONCLUSION: These data indicate that accurately determining past adherence will help identify patients to target to improve their future adherence. These analyses are important for understanding time-varying measures in the clinical setting. PMID- 15806002 TI - Specific IgE and IgG antibody-binding patterns to recombinant cockroach allergens. AB - BACKGROUND: The specificity of serum antibody responses to different cockroach allergens has not been studied. OBJECTIVE: We sought to quantitate serum IgE and IgG antibodies to a panel of purified cockroach allergens among cockroach sensitized subjects. METHODS: IgE antibodies to recombinant cockroach allergens (rBla g 1, rBla g 2, rBla g 4, rBla g 5, and rPer a 7) were measured in sera containing IgE antibodies to Blattella germanica extract (n = 118) by using a streptavidin CAP assay and a multiplex flow cytometric assay. Specific IgG antibodies were determined by using radioimmunoprecipitation techniques. RESULTS: Specific IgE antibodies measured by means of CAP assay and multiplex assay were strongly correlated ( r = 0.8, P < .001). The sum of IgE antibodies (in international units per milliliter) against all 5 allergens equated to IgE antibodies to cockroach extract. Although the prevalence of IgE antibodies was highest for rBla g 2 (54.4%) and rBla g 5 (37.4%), patterns of IgE antibody binding were unique to each subject. Surprisingly, only 16% of cockroach sensitized subjects with IgE antibodies to house dust mite exhibited IgE antibody binding to cockroach tropomyosin (rPer a 7). Specific IgE antibodies were associated with increased IgG antibody levels, although detection of IgG in the absence of IgE was not uncommon. CONCLUSION: The techniques described offer a new approach for defining the hierarchy of purified allergens. IgE antibodies directed against 5 allergens constitute the majority of the IgE antibody repertoire for cockroach. Such distinct patterns of IgE-IgG responsiveness to different cockroach allergens highlight the complexity of B-cell responses to environmental allergens. PMID- 15806005 TI - Ovalbumin-specific IgE modulates ovalbumin-specific T-cell response after repetitive oral antigen administration. AB - BACKGROUND: Some patients outgrow their food allergies even though their serum antigen-specific IgE levels remain high. OBJECTIVE: To elucidate the role of T cells in outgrowing food allergies in the presence of antigen-specific IgE, we tracked antigen-specific T-cell responses after oral antigen administration. METHODS: Ovalbumin (OVA)-specific T-cell receptor (TCR) and OVA-specific IgE transgenic (Tg) mice (OVA-TCR/IgE-Tg) and OVA-specific TCR Tg (OVA-TCR-Tg) mice were fed with high doses of OVA or PBS every other day. After 7 administrations, OVA-specific proliferation and cytokine production of mononuclear cells of the spleen, mesenteric lymph nodes, and Peyer's patches and the number of splenic CD4 + CD25 + T cells were analyzed. RESULTS: Without OVA administration, the splenocytes from OVA-TCR/IgE-Tg mice exhibited a higher proliferative response and produced more IL-4 and IL-10 and less IFN-gamma than those from OVA-TCR-Tg mice. The proliferative responses of the splenocytes from either OVA-TCR/IgE-Tg mice or OVA-TCR-Tg mice fed with OVA were significantly reduced compared with those from PBS-fed mice. The number of OVA-specific TCR + T cells decreased in the spleen from OVA-fed mice, whereas the number of CD4 + CD25 + T cells increased. The suppressed proliferation of splenocytes of OVA-fed mice was partially resumed by neutralization of TGF-beta1, but not of IL-10. CONCLUSION: The presence of OVA-specific IgE modulated the OVA-specific responses of the splenocytes. Irrespective of the presence of OVA-specific IgE, repetitive oral administration of OVA induced tolerance, which seems to be composed of clonal deletion/anergy and TGF-beta1-mediated active suppression. PMID- 15806008 TI - Involvement of human natural killer cells in asthma pathogenesis: natural killer 2 cells in type 2 cytokine predominance. AB - BACKGROUND: T H 2 cells play a pivotal role in the pathogenesis of human asthma. Natural killer (NK) cells are also thought to divide into NK1 and NK2 subsets. OBJECTIVE: Whether NK1 or NK2 cells are involved in asthma remains unclear. METHODS: Triple-color flow cytometry for detecting intracellular cytokine and NK cell surface phenotype from asthmatic patients was used. The NK cell cloning and analysis with RT-PCR for cytokine expression and Western blotting for signal transducer and activator of transcription activation were performed. RESULTS: For the first time, we observed that the ratio of IL-4 + CD56 + NK2 cells in PBMCs of 8 asthmatic patients were higher than in healthy individuals. NK cell clones were then obtained by means of limited dilution, and the average mean of the relative intensity of PCR products for type 2 cytokines significantly increased in the asthmatic patients. Signal transducer and activator of transcription 6, a key transcript factor of type 2 phenotype, was constitutively activated in NK2 clones from asthmatic patients. We cocultured freshly purified NK cells from asthmatic patients with IFN-gamma and anti-IL-4 antibody in the presence of IL-15 and found that the content of IL-4 + NK2 cells significantly decreased after treatment. Interestingly, NK2-biased status in asthmatic patients was reversed when patients recovered from regular therapy. CONCLUSION: The results suggest that the NK2 cell subset is involved in the pathogenesis of asthma. PMID- 15806012 TI - Montelukast protects against bradykinin-induced bronchospasm. PMID- 15806011 TI - Adverse effects of danazol prophylaxis on the lipid profiles of patients with hereditary angioedema. AB - BACKGROUND: Hereditary angioedema (HAE) is a rare disorder caused by the deficiency of the C1-inhibitor gene (C1INH) . Patients experience recurrent bouts of edema, which can occur in almost any region of the body. As regards the treatment of the disease, danazol (an attenuated androgen) is used, among other agents, for long-term prophylaxis. OBJECTIVE: The aim of this study was to investigate the possible adverse effects of danazol on serum lipid profile, as well as to ascertain whether danazol treatment is associated with an increased risk of atherosclerosis. METHODS: Serum concentrations of total cholesterol, high density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides, apolipoprotein A-I, apolipoprotein B-100, and lipoprotein(a) were compared between danazol-treated patients with HAE and 2 control groups (ie, patients who did not receive long-term danazol prophylaxis and untreated healthy subjects). RESULTS: Serum concentrations of HDL ( P = .0002 and P < .0001) and apolipoprotein A-I ( P = .0015 and P < .0001) were significantly lower, whereas LDL ( P = .0129 and P = .0127) and apolipoprotein B-100 ( P = .0456 and P = .0013) were higher in the danazol-treated patients compared with the 2 control groups, respectively. No significant difference was found in total cholesterol, triglyceride, or lipoprotein(a) levels. Patients who received danazol had an 11.6 (95% CI, 2.7-49.7) times higher risk for abnormally low HDL levels and a 4.4 (95% CI, 1.2-16.0) times lower risk for high LDL concentrations. CONCLUSIONS: Our findings indicate that the long-term use of danazol is associated with an increased risk for early atherosclerosis in patients with HAE. Consequently, monitoring of HDL and LDL levels at regular intervals is recommended during follow-up. PMID- 15806014 TI - Missing parvalbumin: implications in diagnostic testing for tuna allergy. PMID- 15806006 TI - Single nucleotide polymorphisms of the IL18 gene are associated with atopic eczema. AB - BACKGROUND: Human IL-18 is an inflammatory cytokine that plays a role in atopic diseases, such as atopic eczema (AE), by enhancing IL-4 and IL-13 production and stimulating the synthesis of IgE. OBJECTIVE: To evaluate associations of polymorphisms in the IL18 gene on chromosome 11q22 with AE, we performed genotyping for single nucleotide polymorphisms (SNPs) in the IL18 gene in 225 patients with AE and 175 healthy control volunteers. METHODS: Genotyping was performed by means of restriction fragment length polymorphism analysis. RESULTS: Analyses revealed significant associations of SNPs +113[t/g] and +127[c/t] in exon 1, -137[g/c] in promoter region 1, and -133[c/g] in promoter region 2 with AE. These associations were not directly dependent on a specific subtype of AE or the concomitant manifestation of allergic rhinitis or asthma. On the functional level, the amount of IL-18 in the supernatants of PBMCs of patients with AE stimulated with Staphylococcus aureus enterotoxin B was significantly higher than that in healthy control subjects. In parallel, the amount of active IL-18 in the sera of patients with AE was enhanced at the exacerbation of their disease. CONCLUSION: In conclusion, our data suggest that SNPs in the IL18 gene might be involved in the development of AE by contributing to a functional dysregulation of the IL-18 production in vivo . PMID- 15806013 TI - Determination of IgG subclasses: a need for standardization. PMID- 15806010 TI - Jun N-terminal kinase is essential for CD40-mediated IgE class switching in B cells. AB - BACKGROUND: CD40 ligation activates nuclear factor kappaB (NF-kappaB) and the mitogen-activated protein kinases p38 and C-Jun N-terminal kinase (JNK) and causes immunoglobulin class-switch recombination (CSR) in B cells. Both NF-kappaB and p38 are important for CD40-mediated CSR. The role of JNK activation in CD40 mediated isotype switching is unknown. OBJECTIVE: We sought to determine the role of JNK activation in CD40-mediated isotype switching. METHODS: Splenic B cells from BALB/c mice were stimulated with anti-CD40 mAb and IL-4 or with soluble CD40 ligand in the presence or absence of SP600125, an anthrapyrazolone inhibitor of JNK. The following events were examined: IgE production by means of ELISA; S(mu) S(epsilon) deletional switch recombination by means of digestion circularization PCR; Cepsilon germline, mature epsilon, and activation-induced deaminase (AID) transcription by means of RT-PCR; and proliferation by tritiated thymidine incorporation and surface expression of CD23, CD54, and CD86 by means of FACS analysis. RESULTS: SP600125 at 10 microM drastically inhibited JNK phosphorylation but had little effect on CD40-mediated p38 phosphorylation and expression of the NF-kappaB dependent genes c-Myc and bcl-xL. SP600125 inhibited IgE synthesis by approximately 88% but had no effect on B-cell proliferation and survival in response to anti-CD40 + IL-4 or on upregulation of CD23, CD54, and CD86 in response to CD40 ligation. Analysis of molecular events involved in IgE class switching revealed that SP600125 had no effect on the expression of C(epsilon) germline and AID transcripts. In contrast, SP600125 severely reduced S(mu)-S(epsilon) switch recombination and expression of mature epsilon transcripts. CONCLUSION: These results demonstrate that JNK activation is essential for CD40-mediated CSR to IgE and suggest that JNK is important for AID activity in B cells. PMID- 15806009 TI - Predictors of immunologic long-term nonprogression in HIV-infected children: implications for initiating therapy. AB - BACKGROUND: Early markers that predict immunologic long-term nonprogression in infants with perinatally acquired HIV infection might assist in subsequent antiretroviral treatment decisions. OBJECTIVES: We sought to identify early markers of immunologic long-term HIV disease nonprogression. METHODS: We analyzed immunologic and virologic characteristics at 1 and 2 months of age in HIV infected children who were enrolled in the Women and Infants Transmission Study and born before 1995, comparing immunologic long-term nonprogressors (ILTNPs; n = 10) with non-ILTNPs (n = 127). ILTNPs were children who survived to 8 years or older with CD4 percentages of 25% or greater and counts of 500 cells/mm 3 or more without receiving highly active antiretroviral therapy. Non-ILTNPs were defined as all other HIV-infected children. Receiver operating characteristic curve analysis was used to assess combined sensitivity and specificity for each of these characteristics and to determine potential threshold values to discriminate between ILTNPs and non-ILTNPs. RESULTS: Characteristics in the first 2 months of life associated with ILTNP status in univariate analysis included higher CD4 percentages, lower CD8 + percentages, lower CD8 + HLA-DR + percentages, and lower HIV-1 RNA PCR values. In receiver operating characteristic analysis CD8 + HLA-DR + percentage had the best combined sensitivity and specificity for discriminating between ILTNPs and non-ILTNPs. CD8 + HLA-DR + percentages of 5% or less predicted ILTNP status with 80% sensitivity and 80% specificity. In multivariate analysis CD8 + HLA-DR+ percentage of 5% or less remained a significant predictor of ILTNP status after adjusting for CD3 + CD4 + percentage and HIV-1 RNA PCR value (odds ratio, 15.4; 95% CI, 1.9-124.7). CONCLUSION: CD8 + HLA-DR + T-lymphocyte percentage of less than 5% at 1 to 2 months of age might be predictive for ILTNP status but should not be used at this time to make treatment-deferral decisions. Immune activation in HIV-infected infants might herald more disease progression. Further study of the use of this subpopulation in early infancy to predict ILTNP status is warranted. PMID- 15806015 TI - Drug presentation to T cells. PMID- 15806016 TI - Does C-2 kinin exist? PMID- 15806018 TI - In vitro synergism of inhaled corticosteroid and long-acting beta2-agonist is not translated clinically in vivo. PMID- 15806036 TI - Genetics and the variability of treatment response in asthma. AB - Pharmacogenetics is the study of how genetic differences influence the variability in patients' responses to therapy. Using specific examples in asthma, I will define the concepts of variability in response to therapy in asthma. I will then use the relationship between specific genetic polymorphisms and the response to beta-adrenergic agonists and leukotriene modifiers to illustrate pharmacogenetic interactions and to acquaint the reader with the applicability of these findings to clinical practice. PMID- 15806038 TI - Gastric reflux is uncommon in acute post-tympanostomy otorrhea. AB - OBJECTIVE: Acute post-tympanostomy otorrhea (APTO) is a common complication of tympanostomy tube placement. APTO has been related primarily to viral upper respiratory infections and external ear contamination. Elevated levels of gastric enzymes have been found in a large proportion of chronic middle ear effusions, implicating gastric reflux (GR) in its pathogenesis. Thus, our objective was to determine whether GR may be a contributing factor in the development of APTO. STUDY DESIGN AND SETTING: Prospective, nonrandomized design. Otorrhea samples were collected from children with APTO. Total pepsinogen concentrations were measured with a commercial ELISA, using a pepsinogen I-specific capture antibody and horseradish peroxidase detection antibody. RESULTS: Twenty-six samples from 24 patients were collected and analyzed. Eight samples demonstrated measurable pepsinogen I, but the measured concentrations, 2-17 mg/L, were below the normal serum reference ranges. CONCLUSIONS: GR does not play a major role in the development of APTO in children. PMID- 15806035 TI - Diversity of asthma: evolving concepts of pathophysiology and lessons from genetics. AB - Asthma is a complex respiratory disorder characterized by reversible airflow obstruction and bronchial hyperreactivity. In recent years, our understanding of the mechanisms underlying allergic asthma has grown, and this has resulted in complex paradigms to define the immunobiology of the disorder. These paradigms conceive of asthma as a disorder with complex genetic and environmental interactions that affect the developing immune system and ultimately result in the episodic release of procontractile mediators, including leukotrienes and prostaglandins, causing susceptible individuals to wheeze. Genetic studies seek to advance our knowledge by identifying the differences in our genetic make-up that cause us to be susceptible. Surprisingly few of the verified asthma genes involve known asthma mediators. This article reviews several of the best established asthma genes, including a disintegrin and metalloprotease 33, dipeptidyl peptidase 10, PHD finger protein 11 (PHF11), and the prostanoid DP1 receptor. Identification of these unsuspected genes is leading to models of asthma pathogenesis that go beyond our conception of asthma as a disease of smooth muscle abnormalities, inflammatory cell presence, and airway structural changes. PMID- 15806037 TI - What the asthma end points we know and love do and do not tell us. AB - Asthma is a disorder characterized by common features of reversible airflow obstruction and bronchial hyperreactivity in the setting of airway mucosal and submucosal inflammation. However, the clinical manifestations are syndromic and not unimodal. There is marked variability in severity of symptoms, natural history, risks of adverse outcomes, pathologic characteristics, and response to therapy. Understanding the relationship between these factors has been complicated by the variability of outcomes and the lack of correlation between them. There is a striking absence of correlation among the pathologic, physiologic, and clinical manifestations of the asthmatic disorders. Lung function tends to correlate poorly with clinical outcomes, and there is only modest correlation between clinical outcomes. Response to therapy is variable both externally (between patients) and internally, depending on which outcome is evaluated. A more complete understanding of the variability of disease and the genetic and environmental causes of the variability likely will change how we approach asthma and its therapy. PMID- 15806020 TI - The discovery of cromolyn sodium and its effect on research and practice in allergy and immunology. PMID- 15806019 TI - Roger Altounyan and the discovery of cromolyn (sodium cromoglycate). PMID- 15806004 TI - Functional assessment of pathogenic IgG subclasses in chronic autoimmune urticaria. AB - BACKGROUND: Chronic urticaria is caused by a complement fixing, IgG antibody directed to the alpha-subunit of the IgE receptor, which is present in 35% to 45% of patients. This autoimmune subgroup can be identified by an autologous skin test or histamine release from human basophils or cutaneous mast cells. However, binding assays do not correlate with these functional assays. We considered the possibility that pathogenic antibody may be present within particular IgG subclasses, which might facilitate development of a binding method that can reliably screen patients. OBJECTIVE: To determine the subclass distribution of IgG antireceptor antibodies on the basis of histamine release, and to assess the possibility that a subclass specific ELISA binding method could be used to screen patients. METHODS: We isolated patient IgG by protein G affinity chromatography and then isolated patient IgG subclasses 1, 2, 3, and 4 by a combination of antibody affinity chromatography and protein A affinity chromatography. The ability of each subclass to activate basophils was assayed by histamine release. RESULTS: Patient IgG subclasses IgG 1 , IgG 3 , and to a lesser degree IgG 4 have antibody capable of activating basophils to release histamine, whereas IgG 2 is inactive. Immunoblot or RAST assay that is subclass-specific does not correlate with histamine release as a result of nonfunctional but binding antibody within IgG subclasses 1, 3, or 4, and complement activation by IgG 1 and IgG 3 . CONCLUSION: Purification of IgG subclasses from patients with chronic urticaria demonstrates functional antibody in IgG 1 and IgG 3 and occasionally IgG 4 . Nonfunctional antibody within IgG 2 plus nonfunctional antibody mixed with functional antibody within IgG 1 , IgG 3 , and IgG 4 and effects of complement are responsible for a lack of correlation of histamine release with binding assays even if subclass-specific. PMID- 15806043 TI - DPOAE-grams in patients with acute tonal tinnitus. AB - OBJECTIVE: To investigate cochlear outer hair cell function in patients with acute tonal tinnitus and normal or near-normal hearing threshold. STUDY DESIGN AND SETTING: Prospective controlled study in an academic tertiary health center. Distortion products of otoacoustic emissions (DPOAE)-grams of 32 ears with acute tonal tinnitus and normal hearing or minimal hearing loss were compared with those of 17 healthy nontinnitus ears. RESULTS: Tinnitus ears exhibited relatively increased amplitudes of DPOAE at high frequencies (4-6.3 kHz) when compared with the group of healthy ears and relatively decreased DPOAE amplitudes at middle frequencies (1650-2400 Hz). Statistically significant ( P < 0.01) increased mean values of DPOAE amplitudes were observed only at a frequency of f2 equal to 4.9 kHz. CONCLUSIONS AND SIGNIFICANCE: These findings suggest an altered functional state of the outer hair cells at a selected high-frequency region of the cochlea in ears with acute tonal tinnitus and normal or near-normal hearing threshold. PMID- 15806044 TI - Management of posttraumatic vertigo. AB - OBJECTIVE: To evaluate patients after blunt trauma of the head, neck, and craniocervical junction (without fractures) with vertigo and to report the results of treatment after extensive diagnostics. STUDY DESIGN: Prospective study of consecutive new cases with vertigo after trauma at different periods of onset. During 2000-2002, 63 patients were examined and treated. SETTING: Regional trauma medical center for the greater Berlin Area, tertiary referral unit. RESULTS: The primary disorders included labyrinthine concussion (18), rupture of the round window membrane (6), and cervicogenic vertigo (12). The secondary disorders included otolith disorders (5), delayed endolymphatic hydrops (12), and canalolithiasis (9). The patients were free of vertigo symptoms (except cervicogenic and otolith disorder) after treatment, which consisted of habituation training, medical and surgical therapy options. The follow-up was 1 year. CONCLUSION: Posttraumatic vertigo can be treated with a high success rate once the underlying disorder has been identified. The extent of the neurotological test battery determines the precision and quality of diagnostics. Surgical measures should be an integral part of treatment modalities if conservative treatment is not effective. SIGNIFICANCE: Minor trauma of the head, neck, and craniocervical junction can have major impact on the vestibular system at different sites. Patients need to be carefully diagnosed, even if the onset of vertigo occurs a few weeks or months after the initial trauma. PMID- 15806040 TI - Mediolateral graft tympanoplasty for anterior or subtotal tympanic membrane perforation. AB - OBJECTIVE: To describe and evaluate the mediolateral graft tympanoplasty for the reconstruction of anterior or subtotal tympanic membrane (TM) perforation. STUDY DESIGN AND SETTING: Retrospective study of 100 patients who underwent the mediolateral graft tympanoplasty at community and tertiary care centers from 1995 to 2001. All patients underwent preoperative and postoperative audiograms. Posterior tympanomeatal flap is elevated same as in the medial (underlay) graft tympanoplasty. Anterior-medial canal skin is elevated down to the annulus. At the annulus, only squamous epithelial layer of TM is elevated up to anterior half of the TM perforation. Temporalis fascia is grafted medial (underlay) to the posterior half of the perforation and lateral (overlay) to the anterior half of the de-epithelialized TM perforation, up to the annulus. Anterior canal skin is rotated to cover the fascia graft and TM perforation as a second-layer closure. Patients were followed for at least 6 months. Outcome was considered successful if the TM is intact. RESULTS: There were 3 failures (97% success rate), attributable to a postoperative infection, anterior blunting, and recurrent cholesteatoma, respectively. There was no significant postoperative hearing loss compared with preoperative hearing. More than 70% of the operated ears had hearing improvement of 0-40 dB (0-10 dB in 19% of ears, 11-20 dB in 44%, 21-30 dB in 7%, and 31-40 dB in 4%) even without ossiculoplasty. With ossiculoplasty using either partial ossicular replacement prosthesis (PORP, 15%) or total ossicular replacement prosthesis (TORP, 11%), there were various degree of hearing improvement from 11 to 30 dB. CONCLUSION AND SIGNIFICANCE: The mediolateral graft method is superior to the traditional medial or lateral graft technique for the reconstruction of large anterior or subtotal TM perforation. This new method should help otologic surgeons to improve outcome of tympanoplasty for anterior or total TM perforation. EBM RATING: C-1. PMID- 15806039 TI - Intratympanic steroid perfusion for refractory sudden sensorineural hearing loss. AB - OBJECTIVE: Patients with sudden sensorineural hearing loss (SSHL) can benefit from systemic steroid therapy. Unfortunately, some patients are not candidates for steroid therapy due to concern over possible complications. Furthermore, not all patients will benefit from steroid administration. This study evaluates the potential benefits and safety of treating patients with SSHL refractory to oral steroids with intratympanic steroid therapy. METHODS: A retrospective case review was performed on all patients who presented with sudden sensorineural hearing loss refractory to oral steroid therapy during the past year. Seventeen patients were identified. All patients underwent intratympanic steroid administration, via MicroWick placement and/or round window catheter placement. RESULTS: Nine patients with sudden sensorineural hearing loss showed an improvement with intratympanic steroid therapy, consisting of MicroWick placement with dexamethasone drop (Decadron) administration for 1-2 weeks and/or round window catheter placement with steroid perfusion. Only one of the patients presenting with hearing loss present for greater than 8 weeks benefited from intratympanic therapy. Complications were few and included tympanic membrane perforation, chronic otitis media, dysequilibrium, and dysguesia. CONCLUSIONS: Intratympanic steroid therapy can be beneficial in treating patients with sudden sensorineural hearing loss refractory to oral steroid use. EBM RATING: C. PMID- 15806041 TI - Safety evaluation of titanium middle ear prostheses at 3.0 tesla. AB - OBJECTIVE: To assess the magnetic resonance imaging (MRI) safety of titanium middle ear prostheses at 3.0 tesla (T). STUDY DESIGN AND SETTING: Titanium middle ear prostheses from 3 commercial vendors were examined for magnetic field interactions at 3 T. Initially, ex vivo studies were performed to test for rotational motion and forward displacement (translational motion) of the prostheses in a static magnetic field. If movement was observed during this screening study, then the prosthesis was tested to determine the translational or rotational force acting upon the prosthesis. In addition to testing for prosthesis displacement, temperature changes of the prostheses were measured to assess for radiofrequency heating during imaging. RESULTS: Twenty-one of the 24 titanium prostheses tested revealed no movement when tested in the 3 T static magnetic field. Three prostheses revealed minimal movement during the screening study. A translational force test (string test) was performed upon these 3 prostheses, and the measured angle of displacement was used to determine the force. This calculated force acting upon each prosthesis was essentially zero. Therefore, we conclude that the magnetic field interaction is negligible. A positive control with a ferromagnetic stainless steel prosthesis demonstrated obvious displacement during the screening study, as well as deflection of the prosthesis by 90 degrees in the translational force test. Last, heating of the titanium prostheses did not occur in the 7 models tested. CONCLUSIONS: Middle ear prostheses made from titanium are safe, neither deflecting nor heating during magnetic resonance examinations conducted at 3 T. EBM RATING: C-1. PMID- 15806048 TI - Local excision procedure for Warthin's tumor of the parotid gland. AB - The aim of this work is to establish a local excision procedure (LEP) and indications of this procedure for Warthin's tumor. Seventy-three patients (82 sides) with Warthin's tumor were studied. Point I was located 1 cm from the intertragal notch in the direction indicated by the notch. Point S was located 5 mm superior to the inferior end of the mandibular angle. The trunk of the facial nerve and the marginal mandibular branch run at the points I and S, respectively. In surgical maneuvers below the I-S line, the marginal mandibular and colli branches may exist within the surgical field, but the trunk and other peripheral branches of the facial nerve will not be encountered. For Warthin's tumor estimated to be below the I-S line (Group A), LEP was used, involving resection of the tumor after locating and dissecting the marginal mandibular and colli branches. For tumors not meeting these criteria (Group B), partial superficial parotidectomy was performed. Results indicated that mean volume of hemorrhage was significantly smaller, and that mean operation time was significantly shorter in Group A than in Group B. Conversely, no significant difference in tumor size or incidence of postoperative facial paresis was identified between the 2 groups. Recurrence has not yet been noted in either group. In conclusion, LEP is useful for Warthin's tumor below the I-S line. This procedure seems applicable not only to Warthin's tumors, but also to other benign parotid tumors in the surgical field below the I-S line, such as pleomorphic adenoma and lymphoepithelial cyst. PMID- 15806042 TI - Reliability of the "Sydney," "Sunnybrook," and "House Brackmann" facial grading systems to assess voluntary movement and synkinesis after facial nerve paralysis. AB - OBJECTIVE: To investigate the extent of within-system reliability and between system correlation for the "Sydney" and "Sunnybrook" systems of grading facial nerve paralysis, and to examine the interobserver reliability and agreement of the "House Brackmann" grading system. STUDY DESIGN: A fixed-effects reliability study in which 6 otolaryngologists viewed videotapes of patients with facial nerve paralysis. SETTING: University and medical Centers. PATIENTS: Patients with unilateral lower motor neurone facial nerve dysfunction greater than 1 year after onset, none of whom had undergone surgical reanimation procedures. Intervention Twenty-one patients with facial nerve paralysis were videotaped while they performed a protocol of facial movements. Six otolaryngologists viewed the videotapes and scored them with the Sydney and Sunnybrook systems, and then gave a House Brackmann grade. MAIN OUTCOME MEASURE: The 3 systems of grading facial nerve paralysis were evaluated and compared with the use of intraclass correlation coefficients, Pearson's weighted kappa, and percentage exact agreement values. RESULTS: The Sydney and the Sunnybrook systems had good intrasystem reliability and high intersystem association for the assessment of voluntary movement. Grading of synkinesis was found to have low reliability both within and between systems. The House Brackmann system had substantial reliability as shown by weighted kappa but had a percentage exact agreement of 44%. CONCLUSIONS: For clinical grading of voluntary movement, there is good correlation between ratings given on the Sydney and Sunnybrook systems, and within each system there is good reliability. The assessment of synkinesis was far less reliable within, and less related between, systems. Although the reliability of the House Brackmann system was found to be high, examination of individual grades revealed some wide variation between trained observers. PMID- 15806047 TI - Sentinel node biopsy as staging tool in a multimodality treatment approach to cancer of the oral cavity and the oropharynx. AB - OBJECTIVES: Feasibility of sentinel lymph node (SLN) biopsy in head and neck cancer as a staging tool embedded in a multimodality regimen including neoadjuvant intraarterial chemotherapy. STUDY DESIGN AND SETTING: 39 patients with oral and anterior oropharyngeal cancer classified N0 by [18 F]FDG-PET underwent SLN scintigraphy. Selective SLN biopsy without elective neck dissection (ND) was performed, immediately followed by radical resection of the primary tumor. Histopathology included step-serial sections and immunocytochemistry. RESULTS: Lymphoscintigraphy detected 104 spots. In 15 patients there was bilateral drainage. 114 SLN were excised due to additional intraoperative discrimination. 95% of visualised SLN could be removed. Histology was positive in 3 patients (8%), all underwent ND which yielded another positive node in 2 cases. Median observation time was 30 months. Two patients (5%) had a neck relapse in combination with a second primary. CONCLUSIONS: SLN biopsy as only surgical staging tool seems to be feasible. SIGNIFICANCE: Method promises reduction of elective ND and morbidity in N0 patients. PMID- 15806045 TI - The bone-anchored hearing aid in children: a surgical and questionnaire follow-up study. AB - OBJECTIVES: The main objectives were to evaluate the surgical techniques and problems seen in children with bone-anchored hearing aids (BAHA) as well as to determine the children's attitudes toward BAHA. STUDY DESIGN: A retrospective study of 41 children with unilateral BAHA was conducted. Surgical recordings were investigated and a questionnaire was distributed. RESULTS: Surgery was performed as a 2-stage procedure. Available bone thickness was measured in 29 patients and found to be on average 2.5 mm. Thin temporal bone necessitated bone augmentation in 8 patients. Of the implants, 70.5% were installed in contact with the dura or the sigmoid sinus. Implant failures were seen in 9.1% and adverse skin reactions appeared in 7.6% of the patients. Twenty-seven patients responded to the questionnaire, 19 were still BAHA users. Overall, these patients were very content with their BAHAs. CONCLUSION: BAHA is a good alternative in children despite limited thickness of the temporal bone. PMID- 15806049 TI - A hystopathological study of thyroarytenoid muscle invasion in early (T1) glottic carcinoma. AB - PATIENTS AND METHODS: Surgical specimens belonging to 16 patients who underwent partial laryngectomy for T1 glottic cancer were subjected to detailed histopathologic examination; 11 of the patients were staged as T1a while 5 were staged as T1b. RESULTS: It was detected that in 11 patients the tumor was confined to the mucosal or submucosal connective layer, and in 5 (31.2%) cases, the tumor invaded the thyroarytenoid (TA) muscle. In cases involving the one third anterior part of the vocal cords and the anterior commissure, a 50% (4/8) TA muscle invasion was detected. DISCUSSION AND CONCLUSION: Our finding TA muscle invasion at the rate of 31.2% in our 16 case series clinically staged as T1 revealed that deep invasion did not always impair the vocal cord mobility. Observing TA muscle invasion at the rate of 50% in cases involving the one-third anterior part of the vocal cord and the anterior commissure suggested that TA muscle invasion in the anterior part did not significantly impair mobility. PMID- 15806046 TI - Long-term effects of repetitive transcranial magnetic stimulation (rTMS) in patients with chronic tinnitus. AB - OBJECTIVES: The pathophysiologic mechanisms of idiopathic tinnitus remain unclear. Recent studies demonstrated focal brain activation in the auditory cortex of patients with chronic tinnitus. Low-frequency repetitive transcranial magnetic stimulation (rTMS) is able to reduce cortical hyperexcitability. STUDY DESIGN: Fusing of the individual PET-scan with the structural MRI-scan (T1, MPRAGE) allowed us to identify exactly the area of increased metabolic activity in the auditory cortex of patients with chronic tinnitus. With the use of a neuronavigational system, this target area was exactly stimulated by the figure 8 shaped magnetic coil. In a prospective study, rTMS (110% motor threshold; 1 Hz; 2000 stimuli/day over 5 days) was performed using a placebo controlled cross-over design. Patients were blinded regarding the stimulus condition. For the sham stimulation a specific sham-coil system was used. Fourteen patients were followed for 6 months. Treatment outcome was assessed with a specific tinnitus questionnaire (Goebel and Hiller). SETTING: Tertiary referral medical center. RESULTS: Increased metabolic activation in the auditory cortex was verified in all patients. After 5 days of verum rTMS, a highly significant improvement of the tinnitus score was found whereas the sham treatment did not show any significant changes. The treatment outcome after 6 months still demonstrated significant reduction of tinnitus score. CONCLUSION: These preliminary results demonstrate that neuronavigated rTMS offers new possibilities in the understanding and treatment of chronic tinnitus. PMID- 15806050 TI - The utility of serum PTH assessment 24 hours after total thyroidectomy. AB - BACKGROUND: Hypocalcemia is the most frequent complication following total thyroidectomy. This prospective study examines the predictive value of parathyroid hormone (PTH) levels measured 24 hours after surgery. MATERIAL AND METHODS: A total of 1006 consecutive patients (mean age, 54.8 years; female/male ratio, 4/1) underwent total thyroidectomy for benign or malignant thyroid from January 1995 to November 2003. Serum calcium, phosphorus, and PTH were measured preoperatively and at 24 hours after surgery. All patients underwent preoperative examination to assess cord motility. RESULTS: A total of 253 (25.1%) patients presented with hypocalcemia demonstrated by clinical and laboratory findings. In 101 cases the hypocalcemic syndrome manifested after 24 to 36 hours whereas in 5 of 101 cases, symptom onset was between 48 and 72 hours. Serum calcium levels lower than 7.5 mg/dL were recorded in all the 101 cases. In 239 of 253 cases serum calcium returned to normal values within 7 days following surgery. PTH at 24 hours was below normal levels in 49 of the 101 patients but was within normal limits in 52 cases. The incidence of hypocalcemia was higher in patients undergoing surgery for malignant thyroid ( P < 0.05). CONCLUSIONS: We do not consider PTH levels at 24 hours postoperatively as predictive of hypocalcemia. PMID- 15806017 TI - Laboratory diagnostics for hereditary angioedema: an economic, evidence-based standpoint. PMID- 15806051 TI - Primary parapharyngeal space tumors in a Mexican cancer center. AB - OBJECTIVE: To describe clinical and demographic characteristics of the parapharyngeal space tumors and assess surgical approaches used to treat them at our institution. METHODS: A retrospective and descriptive study of the parapharyngeal space tumors, excluding paragangliomas, treated from June 1991 to October 2002 in a cancer center. The study population included 21 patients, 8 men and 13 women, average age of 41 years (range, 20 to 70 years). Fine needle biopsy was done in 5 (24%) patients. Computed tomography (CT) was performed in all patients, and only a few required magnetic resonance image (MRI). RESULTS: Surgical approaches included transcervical alone or in combination with parotidectomy, transoral, or transmandibular (mandibular swing) approach. Laminectomy and segmentary approaches were also performed in 1 patient each. Sixteen (76%) patients had benign lesions and 5 (24%) had malignant tumors. Neurogenic tumors represented 57% of all tumors. Mean tumor size was of 6.7 cm (range, 3 to 11 cm). Six (29%) patients received adjuvant radiotherapy. Complications occurred in 6 (29%) patients, 4 (19%) of which were nervous injuries associated with peripheral nerve sheath tumors. Median disease-free follow-up survival was 33 months (range, 2 to 184 months) despite being an heterogeneous group of histologies. CONCLUSION: Parapharyngeal space is a rare location for head and neck tumors. Cervical approach should be the first choice for large tumors; transoral approach is reserved for tumors less than 3 cm. Conversion to mandibular swing approach when the cervical approach is not offering proper exposure for tumor resection is indicated. Preoperative histologic diagnosis is not required. Nevertheless, CT scan should always be performed in order to exclude paragangliomas, distinguish prestyloid from poststyloid lesions, and to assess the extension of the tumor as well as its relationship with adjacent structures. PMID- 15806052 TI - Bone regeneration in the rat mandible with bone morphogenetic protein-2: a comparison of two carriers. AB - OBJECTIVE: To compare mandibular bone regeneration with bone morphogenetic protein-2 (BMP-2) delivered with two carriers: a hyaluronic acid polymer (HY), and a collagen carrier complexed with calcium hydroxyapatite and tricalcium phosphate (collagen/HA/TCP). STUDY DESIGN: Defects were created in the bilateral mandibular bodies of 16 Sprague-Dawley rats. The defects were filled with the HY carrier, the HY carrier loaded with BMP-2, the collagen/HA/TCP carrier, or the collagen/HA/TCP carrier loaded with BMP-2. Animals were euthanatized after 6 weeks, and the hemi-mandibles were analyzed histomorphologically. RESULTS: Specimens containing BMP-2 had significantly larger new bone and marrow volumes than control specimens. Specimens in the hyaluronan/BMP-2 group tended to have larger volumes of new bone and osteoid than collagen/HA/TCP/BMP-2 specimens, though these differences were not statistically significant. CONCLUSION: The HY and collagen/HA/TCP carriers had comparable efficacy for bone healing with BMP-2. SIGNIFICANCE: Bone morphogenetic proteins can be delivered with commercially available alloplasts as osteogenic bone substitutes for the repair of craniofacial bone defects. EBM RATING: B-2. PMID- 15806053 TI - Use of mitomycin C for treatment of keloid: a preliminary report. AB - OBJECTIVES: Mitomycin C (MC) is an anti-neoplastic agent with an anti proliferative effect on fibroblasts. We set out to evaluate the effect of MC application following keloid excision. STUDY DESIGN AND SETTING: Patients with keloid in a tertiary referral center were enrolled in a prospective study. The scar was excised and prior to skin closure, a pledget with 1 cc of MC 0.4 mg/mL was applied for 5 minutes. Patient satisfaction and keloid thickness were assessed. RESULTS: All patients were satisfied with the results, although complete disappearance of the keloid was evident only in two. Keloid thickness was measured and ranged from 5 to 26 mm. Following surgery and treatment at 2 months thickness ranged from 0 to 8 mm. CONCLUSIONS: MC application following scar resection appears to be effective in treatment of keloid. SIGNIFICANCE: As no specific effective treatment for keloid is currently available, utilization of this readily available therapeutic agent may improve treatment outcome. PMID- 15806054 TI - Endoscopic surgery of inverted papillomas under image guidance--a prospective study of 42 consecutive cases at a Danish university clinic. AB - BACKGROUND: To evaluate the feasibility of endoscopic surgery with image guidance in the treatment of inverted papillomas. STUDY DESIGN AND SETTING: This prospective cohort study comprised 42 consecutive patients with biopsy-confirmed inverted papillomas that were diagnosed between 1998 and 2003. All patients were treated by the first author (C.B.). Image guidance based on preacquired CT scans of the patients was used to assist the surgeon aiming at endoscopic resection of inverted papilloma. The success of the surgery was judged primarily by the recurrence rate and the treatment morbidity. RESULTS: The study group consisted of 30 males and 12 females with a median age of 61 years. The follow-up period ranged from 9 months to 69 months (median, 37 months). The only additional procedure performed was the Caldwell-Luc procedure (8 cases). Recurrence was diagnosed in 10 cases (24%), all in the original tumor site. Eight of these had undergone previous surgery for inverted papilloma. The recurrences were predominantly located in the maxillary or in the frontal sinus (8 cases). In 2 cases, the recurrence was simply removed endoscopically while performing the biopsy procedure. All recurrences were identified within the first 9 months. Associated malignancy was not shown. No severe complications occurred. CONCLUSIONS: A treatment based on endoscopic resection with image guidance appears to offer a safe treatment modality of inverted papilloma with insignificant morbidity. Predominantly cases with nonmedial involvement of the maxillary sinus still require a supplement with the Caldwell Luc procedure. Although all the recurrences were found within 9 months postoperatively, later recurrences cannot be excluded. Long-term follow-up is recommended. PMID- 15806055 TI - Use of mometasone furoate aqueous nasal spray in the treatment of rhinitis medicamentosa: an experimental study. AB - OBJECTIVE: We aimed to investigate, histopathologic changes in the nasal mucosa of guinea pig's after prolonged administration of oxymetazoline and the development of rhinitis medicamentosa, and the efficacy of mometasone furoate aqueous nasal spray and saline in reversing the ultrastructural changes attributable to rhinitis medicamentosa. METHODS: In the study, 24 male guinea pigs (500 to 600 gr) were used. Oxymetazolin (0.05%) was sprayed into the nasal cavities of the guinea pigs 3 times daily for 8 weeks. At the end of this period, 6 guinea pigs were killed and examined to make sure that the animals had developed rhinitis medicamentosa. The remaining guinea pigs were randomly divided into 3 groups. In the first group, one spray-puff of 0.05% mometasone furoate aqueous nasal spray (50 microg) was applied twice daily for 14 days. In the second group, saline solution (0.9% NaCl) was applied twice daily for 14 days. No treatment was performed in the third group. At the end of the treatment period, nasal mucosal changes were evaluated by light microscopy and electron microscopy. RESULTS: After oxymetazolin application for 8 weeks, the main histologic changes were edema, congestion, proliferation of subepithelial glands, and squamous cell metaplasia. After topical mometasone furoate aqueous spray application for 2 weeks, the edema fluid was found to diminish markedly. In the saline and no treatment groups, edema and congestion continued. In these groups of guinea pigs, fibrosis has been seen in the nasal mucosa. CONCLUSION: We found that mometasone furoate nasal spray was effective against experimentally induced rhinitis medicamentosa in guinea pigs. Mometasone furoate nasal spray may have value in the treatment of patients with rhinitis medicamentosa. PMID- 15806056 TI - Expression of interleukins in patients with nasal polyposis. AB - OBJECTIVE: To correlate the levels of interleukins 1beta, 3, 4, and 5 before and after surgery and compare the levels between patients with and without recurrence of nasal polyposis. STUDY DESIGN AND SETTING: Thirty-nine patients with NP were selected, 13 of them allergic and 26 nonallergic. A control group of 11 individuals was also studied. The concentrations of interleukins were measured by enzyme-linked immunosorbent assay. RESULTS: There was a higher incidence of NP after the fourth decade of life and among men. The clinical symptoms were similar in both groups of patients with nasal polyposis and characterized by nasal obstruction and anosmia. A significant reduction of all interleukins studied was observed after surgical treatment. CONCLUSION: Levels of interleukins 1beta, 3, 4, and 5 were significantly reduced after surgery and the levels of interleukins 1beta and 5 were significantly lower in patients without recurrence of nasal polyposis after surgery when compared to those with recurrence. PMID- 15806057 TI - Ultrastructural detection of nitric oxide in human nasal mucosa. AB - OBJECTIVE: Nasal vasculature and seromucous glands are exposed to complex mechanisms influenced by external as well as internal stimuli. In addition to classic and peptidergic neurotransmitters, nitric oxide (NO) was increasingly found to be important in the control of various physiologic functions. NO modulates nasal immunology, influences macrophage activity, and has antiviral and bacteriostatic properties. The aim of this study was to show the localization of nitric oxide synthases (NOS) I and III in the normal human nasal mucosa by using immunoelectron microscopical techniques. STUDY DESIGN: Specimens of noninflammed inferior turbinates from 35 patients who underwent nasal surgery were fixed in phosphate-buffered glutaraldehyde. After dehydration, incubation in unicryl and polymerization, ultrathin sections were cut. Primary antibodies against NOS I and III were applied and the immunocomplexes were visualized by an immunocytochemical staining-technique using gold-labeled antibodies. Immunostained structures were photodocumented using a transmission electron microscope. RESULTS: NOS immunoreactive nerve fibers were mainly co-located in parasympathetic nerves in the adventitia of arterial vessels and in periglandular axons. Electron microscopy showed that NOS-positive axons were in close contact with acinus cells. A strong NOS III-immunoreactivity was found in endothelial cells of capillaries near the glands as well as in arterial vessels. Furthermore, immunoreaction products were deposited throughout the cytoplasm of fibroblasts. CONCLUSION: Nitric oxide in nerve fibers, seromucous glands, and endothelial cells of capillaries and arterial vessels suggest that NO takes part in the regulation of physiologic processes of the human nasal mucosa. NOS was co localized in parasympathetic nerves and plays a role in the neurotransmission and neuromodulation of the vascular tone and glandular secretion. Arteries showed a distinctly developed nitrergic innervation and endothelial accumulation. The NO production in axons of the adventitia and in the endothelium of arteries demonstrated that these vessels are influenced by a dual NO system. NO could mainly act on these structures with vasodilatatory effects. Finally, NO would be able to influence the functions of perivascular fibroblasts. PMID- 15806058 TI - Eustachian tube dysfunction in allergic rhinitis. AB - OBJECTIVE: To assess eustachian tube function in patients with allergic rhinitis and compare them with a control group. STUDY DESIGN AND SETTING: Tympanometry was performed in 130 patients (260 ears), divided into 2 groups: 80 cases with allergic rhinitis and 50 healthy controls. Cases underwent skin hypersensitivity tests. RESULTS: Cases, age 21.1 +/- 14.9; Controls, age 23.9 +/- 15.6. Most frequent skin hypersensitivity: Dermatophagoides pt (62%), Zea Maiz (44%), and Cockroach (37%). Tympanometry of cases showed negative values of peak tympanometric pressure in both children and adults (P < or = 0.05). Among children under 11 years of age, 15.5% tympanograms showed abnormal curves (13% C curves and 3% B curves); among the control group only normal curves were found (type A). CONCLUSION: Allergic rhinitis patients have a higher risk of eustachian tube dysfunction, particularly during childhood. Tympanometry is a noninvasive, readily available procedure that may be useful in these patients to prevent chronic middle-ear disease. PMID- 15806060 TI - Effects of genioglossal muscle advancement on speech: an acoustic study of vowel sounds. AB - OBJECTIVE: The effects of the genioglossal muscle advancement on phonetic quality of speech were studied analyzing the acoustic features of vowel sounds. STUDY DESIGN AND SETTING: The study group consisted of 5 men suffering from partial upper airway obstruction during sleep. To prevent tongue base collapse, genioglossal muscle advancement was made with chin osteotomy without hyoid myotomy and suspension. The speech material consisted of 8 vowels produced in sentence context repeated 10 times before the operation, and 10 days and 6 weeks after the operation. The acoustic features of vowels were analyzed. RESULTS: The operation had no significant effects on vowel quality. Only for 2 of the subjects the pitches changed systematically due to the operation. CONCLUSION: According to the acoustic analysis, genioglossal muscle advancement with chin osteotomy has no effects on vowel production. Some short-term changes were observed, but these changes were highly individual. SIGNIFICANCE: The operation seems to have no potential to change vowel production. PMID- 15806059 TI - Multilevel temperature-controlled radiofrequency for obstructive sleep apnea: extended follow-up. AB - OBJECTIVE: To determine long-term effectiveness of multilevel (tongue and palate) temperature-controlled radiofrequency tissue ablation (TCRFTA) for patients with obstructive sleep apnea syndrome (OSAS). STUDY DESIGN AND SETTING: Prospective, 2 institution case series. Twenty-nine subjects with mild to moderate OSAS and who were at least 1 year from completion of multilevel TCRFTA were included, representing a subset of subjects who were enrolled in a previously published controlled trial. Exclusion criteria for this extended follow-up study included any additional treatment for OSAS after completion of TCRFTA. RESULTS: Median follow-up was 23 months. Daytime sleepiness and OSAS-related quality of life were significantly improved at extended follow-up (both P < 0.001). Median reaction time testing and apnea-hypopnea index (AHI) were also significantly improved at long-term follow-up (P = 0.03 and 0.01). Body mass index was unchanged (P = 0.94). CONCLUSIONS: Multilevel TCRFTA treatment of mild to moderate OSAS resulted in prolonged improvement in daytime somnolence, OSAS-related quality of life, psychomotor vigilance, and AHI in this group of subjects at extended follow-up. PMID- 15806061 TI - The role of adjustment of expiratory effort in the control of vocal intensity: clinical assessment of phonatory function. AB - OBJECTIVE: To determine the role of the adjustment of expiratory effort in the control of vocal intensity. STUDY DESIGN: An intensity-loading test was performed by using the airway interruption method. Three groups of subjects were used: a control group thought to resemble normal vocal fold closure, a group of patients with Reinke's edema thought to represent increased mass at the level of the vocal folds, and a group with vocal fold paralysis that was thought to represent a group with lack of adequate vocal fold closure. RESULTS: In the control group, expiratory lung pressure and airway resistance slightly increased. In the patients with Reinke's edema, expiratory lung pressure, and airway resistance significantly increased. In this group, the voice intensity was controlled by laryngeal adjustment, but a greater expiratory effort was needed because of a greater increase in glottal resistance. In the patients with vocal cord paralysis, airway resistance did not increase even with a high-intensity voice. Vocal intensity was controlled by expiratory effort. CONCLUSIONS: If there is sufficient ability for laryngeal adjustment, vocal intensity is controlled primarily by laryngeal adjustment and by expiratory adjustment in response to increased glottal resistance. However, vocal intensity is controlled by expiratory effort when laryngeal adjustment ability is poor. PMID- 15806062 TI - Measuring quality of life in preschool children with sore throats and otitis media using the TAPQOL questionnaire. AB - OBJECTIVES: TNO-AZL Preschool Quality of Life Questionnaire (TAPQOL) is one of the few instruments designed to assess health-related quality of life in preschool children but its applicability to otolaryngology is unknown. STUDY DESIGN AND SETTING: We studied a consecutive series of children aged 1 to 5 years referred to hospital with recurrent sore throats, recurrent acute otitis media, or otitis media with effusion. RESULTS: TAPQOL domain scores were not influenced by age, sex, or socio-economic class, but correlated with markers of disease severity (frequency of sore throat or pyrexia, time off school), ear-related handicap (assessed with the OM6 questionnaire), and other measures of health related quality of life (visual analogue scale, 5-point rating scale and the Health Utilities Index mark III). Comparison with published data from healthy children suggests that these common otolaryngologic problems have a large impact on a child's quality of life. CONCLUSIONS: TAPQOL seems to be appropriate for use in this context. Marked ceiling effects in some domains, however, may limit their sensitivity. PMID- 15806063 TI - Imperforate submandibular duct. PMID- 15806064 TI - Pediatric subglottic granular cell myoblastoma. PMID- 15806065 TI - Stomatococcus mucilaginosis infection leading to early cervical necrotizing fasciitis. PMID- 15806066 TI - Tracheal paraganglioma presenting with acute airway obstruction. PMID- 15806067 TI - Metastatic renal cell carcinoma to the mandible. PMID- 15806068 TI - Parotitis due to ritodrine tocolytic treatment for preterm labor. PMID- 15806069 TI - Migrating foreign body into the common carotid artery. PMID- 15806070 TI - Fifty-four wooden toothpicks in the nasal cavity. PMID- 15806071 TI - Internal carotid pseudoaneurysm within a parapharyngeal infection: an infrequent complication of difficult diagnosis. PMID- 15806074 TI - One gene, one complex neurological disorder? PMID- 15806075 TI - A therapy that'll put hair in your ears. PMID- 15806078 TI - Not your normal redleg. PMID- 15806081 TI - NIH updates regulations on conflict of interest. PMID- 15806082 TI - New guidance on prompt reporting to OLAW under PHS Policy. PMID- 15806083 TI - Training and feasibility protocols: acceptable under current laws and standards? Details missing. PMID- 15806084 TI - Training and feasibility protocols: acceptable under current laws and standards? PMID- 15806085 TI - Training and feasibility protocols: acceptable under current laws and standards? Too much ambiguity. PMID- 15806086 TI - Training and feasibility protocols: acceptable under current laws and standards? A bridge too far. PMID- 15806087 TI - Perianal mass in a female transgenic mouse. PMID- 15806089 TI - Breeding genetically modified rodents: tips for tracking and troubleshooting reproductive performance. AB - Infertility or reduced fertility in genetically modified mouse strains represents a serious bottleneck for planned research projects. The authors describe simple methods for troubleshooting poor reproductive performance in breeding colonies and introduce various means of assisted reproduction that might be used to 'rescue' infertile lines. PMID- 15806090 TI - The role of the veterinary staff in mouse breeding colony management. AB - The rapid increase in the production and use of transgenic mice has been a boon for biomedical research and a challenge for the animal care and use programs responsible for providing housing and medical care to these animals. The authors suggest ways in which the veterinary staff can successfully organize and manage transgenic mouse breeding programs to reduce uncontrolled breeding and the problems associated with it. PMID- 15806091 TI - Archiving mouse strains by cryopreservation. AB - A great deal of time and energy goes into the creation of each new line of transgenic mice; established lines are expensive and labor-intensive to maintain. Archiving of mice by cryopreservation of germ cells or embryos represents a means to free up facility space, while protecting the line from loss due to environmental disasters, genetic drift, or infectious disease. The author reviews the available cryopreservation techniques and presents considerations for setting up a cryopreservation facility. PMID- 15806092 TI - Reestablishment of a transgenic rabbit line by artificial insemination using cryopreserved semen. AB - The production of recombinant proteins in the milk of transgenic animals is an alternative to traditional cell culture methodology. Transgenic rabbits can serve in the small-scale production of recombinant proteins, underscoring the need to maintain valuable transgenic lines. In this study, the authors used cryopreserved transgenic rabbit semen to artificially inseminate does, demonstrating the utility of this method for the reestablishment of a transgenic rabbit herd. PMID- 15806093 TI - A primer on rodent identification methods. AB - Identifying laboratory rodents as a group may be sufficient if all the animals are to receive the same treatment and/or manipulation and if individual variations in response are not to be recorded separately. However, it is frequently necessary to be able to differentiate between individual animals used in a study. Identification of individual animals may also be necessary to maintain health records and to properly manage colonies of laboratory animals in compliance with regulatory agencies and research protocol requirements. PMID- 15806094 TI - Insights into host responses against pathogens from transcriptional profiling. AB - DNA microarrays have allowed us to monitor the effects of pathogens on host-cell gene expression programmes in great depth and on a broad scale. The comparison of results that have been generated by these studies is complex, and such a comparison has not previously been attempted in a systematic manner. In this review, we have collated and compared published transcriptional-profiling data from 32 studies that involved 77 different host-pathogen interactions, and have defined a common host-transcriptional-response. We outline gene expression patterns in the context of Toll-like receptor and pathogen-mediated signalling pathways, and summarize the contributions that transcriptional-profiling studies have made to our understanding of the infectious disease process. PMID- 15806095 TI - Amyloids--a functional coat for microorganisms. AB - Amyloids are filamentous protein structures approximately 10 nm wide and 0.1-10 mum long that share a structural motif, the cross-beta structure. These fibrils are usually associated with degenerative diseases in mammals. However, recent research has shown that these proteins are also expressed on bacterial and fungal cell surfaces. Microbial amyloids are important in mediating mechanical invasion of abiotic and biotic substrates. In animal hosts, evidence indicates that these protein structures also contribute to colonization by activating host proteases that are involved in haemostasis, inflammation and remodelling of the extracellular matrix. Activation of proteases by amyloids is also implicated in modulating blood coagulation, resulting in potentially life-threatening complications. PMID- 15806096 TI - Opinion: Bacterial toxins and cancer--a case to answer? AB - Since the discovery that Helicobacter pylori infection leads to gastric cancer, other chronic bacterial infections have been shown to cause cancer. The bacterial and host molecular mechanisms remain unclear. However, many bacteria that cause persistent infections produce toxins that specifically disrupt cellular signalling to perturb the regulation of cell growth or to induce inflammation. Other bacterial toxins directly damage DNA. Such toxins mimic carcinogens and tumour promoters and might represent a paradigm for bacterially induced carcinogenesis. PMID- 15806097 TI - Highly efficient endogenous human gene correction using designed zinc-finger nucleases. AB - Permanent modification of the human genome in vivo is impractical owing to the low frequency of homologous recombination in human cells, a fact that hampers biomedical research and progress towards safe and effective gene therapy. Here we report a general solution using two fundamental biological processes: DNA recognition by C2H2 zinc-finger proteins and homology-directed repair of DNA double-strand breaks. Zinc-finger proteins engineered to recognize a unique chromosomal site can be fused to a nuclease domain, and a double-strand break induced by the resulting zinc-finger nuclease can create specific sequence alterations by stimulating homologous recombination between the chromosome and an extrachromosomal DNA donor. We show that zinc-finger nucleases designed against an X-linked severe combined immune deficiency (SCID) mutation in the IL2Rgamma gene yielded more than 18% gene-modified human cells without selection. Remarkably, about 7% of the cells acquired the desired genetic modification on both X chromosomes, with cell genotype accurately reflected at the messenger RNA and protein levels. We observe comparably high frequencies in human T cells, raising the possibility of strategies based on zinc-finger nucleases for the treatment of disease. PMID- 15806098 TI - Electroreceptor neuron dynamics shape information transmission. AB - The gymnotiform weakly electric fish Apteronotus leptorhynchus can capture prey using electrosensory cues that are dominated by low temporal frequencies. However, conventional tuning curves predict poor electroreceptor afferent responses to low-frequency stimuli. We compared conventional tuning curves with information tuning curves and found that the latter predicted substantially improved responses to these behaviorally relevant stimuli. Analysis of receptor afferent baseline activity showed that negative correlations reduced low frequency noise levels, thereby increasing information transmission. Multiunit recordings from receptor afferents showed that this increased information transmission could persist at the population level. Finally, we verified that this increased low-frequency information is preserved in the spike trains of central neurons that receive receptor afferent input. Our results demonstrate that conventional tuning curves can be misleading when certain noise reduction strategies are used by the nervous system. PMID- 15806099 TI - Genetically enhanced cows resist intramammary Staphylococcus aureus infection. AB - Mastitis, the most consequential disease in dairy cattle, costs the US dairy industry billions of dollars annually. To test the feasibility of protecting animals through genetic engineering, transgenic cows secreting lysostaphin at concentrations ranging from 0.9 to 14 micrograms/ml [corrected] in their milk were produced. In vitro assays demonstrated the milk's ability to kill Staphylococcus aureus. Intramammary infusions of S. aureus were administered to three transgenic and ten nontransgenic cows. Increases in milk somatic cells, elevated body temperatures and induced acute phase proteins, each indicative of infection, were observed in all of the nontransgenic cows but in none of the transgenic animals. Protection against S. aureus mastitis appears to be achievable with as little as 3 micrograms/ml [corrected] of lysostaphin in milk. Our results indicate that genetic engineering can provide a viable tool for enhancing resistance to disease and improve the well-being of livestock. PMID- 15806100 TI - Extracellular secretion of polypeptides using a modified Escherichia coli flagellar secretion apparatus. AB - We developed a modified flagellar type III secretion apparatus to secrete heterologous polypeptides into the growth medium of Escherichia coli. The secretion was facilitated by fusing the 173-bp untranslated DNA fragment upstream of the gene fliC (encoding flagellin) as well as a transcriptional terminator from fliC, into the gene encoding the polypeptide of interest. The polypeptides secreted into the growth medium at concentrations ranging from 1 to 15 mg/l were from Campylobacter jejuni (262 residues in length), Streptococcus pneumoniae (434 residues), Staphylococcus aureus (115 residues), and N-terminal FliC hybrid proteins, for example, the eukaryotic green fluorescent protein (238 residues). The expressed proteins represented >50% of total secreted protein. Previously reported protein yields from extracellular secretion of foreign proteins in E. coli have been low, approximately 100 microg/l. The strengths of our method are the concentration and purity of the secreted proteins and its versatility with regard to the proteins' length and origin. PMID- 15806101 TI - A gene expression map of Arabidopsis thaliana development. AB - Regulatory regions of plant genes tend to be more compact than those of animal genes, but the complement of transcription factors encoded in plant genomes is as large or larger than that found in those of animals. Plants therefore provide an opportunity to study how transcriptional programs control multicellular development. We analyzed global gene expression during development of the reference plant Arabidopsis thaliana in samples covering many stages, from embryogenesis to senescence, and diverse organs. Here, we provide a first analysis of this data set, which is part of the AtGenExpress expression atlas. We observed that the expression levels of transcription factor genes and signal transduction components are similar to those of metabolic genes. Examining the expression patterns of large gene families, we found that they are often more similar than would be expected by chance, indicating that many gene families have been co-opted for specific developmental processes. PMID- 15806102 TI - A genomic screen in yeast implicates kynurenine 3-monooxygenase as a therapeutic target for Huntington disease. AB - Huntington disease is a fatal neurodegenerative disorder caused by expansion of a polyglutamine tract in the protein huntingtin (Htt), which leads to its aggregation in nuclear and cytoplasmic inclusion bodies. We recently identified 52 loss-of-function mutations in yeast genes that enhance the toxicity of a mutant Htt fragment. Here we report the results from a genome-wide loss-of function suppressor screen in which we identified 28 gene deletions that suppress toxicity of a mutant Htt fragment. The suppressors are known or predicted to have roles in vesicle transport, vacuolar degradation, transcription and prion-like aggregation. Among the most potent suppressors was Bna4 (kynurenine 3 monooxygenase), an enzyme in the kynurenine pathway of tryptophan degradation that has been linked directly to the pathophysiology of Huntington disease in humans by a mechanism that may involve reactive oxygen species. This finding is suggestive of a conserved mechanism of polyglutamine toxicity from yeast to humans and identifies new candidate therapeutic targets for the treatment of Huntington disease. PMID- 15806103 TI - Maternal segregation of the Dutch preeclampsia locus at 10q22 with a new member of the winged helix gene family. AB - Preeclampsia is a pregnancy-associated disease with maternal symptoms but placental origin. Epigenetic inheritance is involved in some populations. By sequence analysis of 17 genes in the 10q22 region with maternal effects, we narrowed the minimal critical region linked with preeclampsia in the Netherlands to 444 kb. All but one gene in this region, which lies within a female-specific recombination hotspot, encode DNA- or RNA-binding proteins. One gene, STOX1 (also called C10orf24), contained five different missense mutations, identical between affected sisters, cosegregating with the preeclamptic phenotype and following matrilineal inheritance. Four STOX1 transcripts are expressed in early placenta, including invasive extravillus trophoblast, generating three different isoforms. All contain a winged helix domain related to the forkhead (FOX) family. The largest STOX1 isoform has exclusive nuclear or cytoplasmic expression, indicating activation and inactivation, respectively, of the PI3K-Akt-FOX pathway. Because all 38 FOX proteins and all 8 STOX1 homologs have either tyrosine or phenylalanine at position 153, the predominant Y153H variation is highly mutagenic by conservation criteria but subject to incomplete penetrance. STOX1 is a candidate for preeclampsia controlling polyploidization of extravillus trophoblast. PMID- 15806104 TI - Combinatorial microRNA target predictions. AB - MicroRNAs are small noncoding RNAs that recognize and bind to partially complementary sites in the 3' untranslated regions of target genes in animals and, by unknown mechanisms, regulate protein production of the target transcript. Different combinations of microRNAs are expressed in different cell types and may coordinately regulate cell-specific target genes. Here, we present PicTar, a computational method for identifying common targets of microRNAs. Statistical tests using genome-wide alignments of eight vertebrate genomes, PicTar's ability to specifically recover published microRNA targets, and experimental validation of seven predicted targets suggest that PicTar has an excellent success rate in predicting targets for single microRNAs and for combinations of microRNAs. We find that vertebrate microRNAs target, on average, roughly 200 transcripts each. Furthermore, our results suggest widespread coordinate control executed by microRNAs. In particular, we experimentally validate common regulation of Mtpn by miR-375, miR-124 and let-7b and thus provide evidence for coordinate microRNA control in mammals. PMID- 15806105 TI - Regulation of T cell receptor alpha gene assembly by a complex hierarchy of germline Jalpha promoters. AB - Assembly of the gene encoding T cell receptor alpha (Tcra) is characterized by an orderly progression of primary and secondary V(alpha)-to-J(alpha) recombination events across the J(alpha) array, but the targeting mechanisms responsible for this progression are mostly unknown. Studies have shown that the T early-alpha promoter is important in targeting primary Tcra rearrangements. We found that T early-alpha and a previously unknown promoter associated with J(alpha)49 targeted primary recombination to discrete sets of constant alpha region (C(alpha))-distal J(alpha) segments and together directed nearly all normal primary recombination events. Furthermore, deletion of the T early-alpha promoter activated previously suppressed downstream promoters and stimulated primary rearrangement to centrally located J(alpha) segments. Central promoter derepression also occurred after primary rearrangement, thereby providing a mechanism to target secondary recombination events. PMID- 15806106 TI - Suppression of allergic inflammation by the prostaglandin E receptor subtype EP3. AB - Prostaglandins, including PGD(2) and PGE(2), are produced during allergic reactions. Although PGD(2) is an important mediator of allergic responses, aspirin-like drugs that inhibit prostaglandin synthesis are generally ineffective in allergic disorders, suggesting that another prostaglandin-mediated pathway prevents the development of allergic reactions. Here we show that such a pathway may be mediated by PGE(2) acting at the prostaglandin E receptor EP3. Mice lacking EP3 developed allergic inflammation that was much more pronounced than that in wild-type mice or mice deficient in other prostaglandin E receptor subtypes. Conversely, an EP3-selective agonist suppressed the inflammation. This suppression was effective when the agonist was administered 3 h after antigen challenge and was associated with inhibition of allergy-related gene expression. Thus, the PGE(2)-EP3 pathway is an important negative modulator of allergic reactions. PMID- 15806107 TI - A Brownian motor mechanism of translocation and strand separation by hepatitis C virus helicase. AB - Helicases translocate along their nucleic acid substrates using the energy of ATP hydrolysis and by changing conformations of their nucleic acid-binding sites. Our goal is to characterize the conformational changes of hepatitis C virus (HCV) helicase at different stages of ATPase cycle and to determine how they lead to translocation. We have reported that ATP binding reduces HCV helicase affinity for nucleic acid. Now we identify the stage of the ATPase cycle responsible for translocation and unwinding. We show that a rapid directional movement occurs upon helicase binding to DNA in the absence of ATP, resulting in opening of several base pairs. We propose that HCV helicase translocates as a Brownian motor with a simple two-stroke cycle. The directional movement step is fueled by single stranded DNA binding energy while ATP binding allows for a brief period of random movement that prepares the helicase for the next cycle. PMID- 15806108 TI - Structure of the SWI2/SNF2 chromatin-remodeling domain of eukaryotic Rad54. AB - SWI2/SNF2 chromatin-remodeling proteins mediate the mobilization of nucleosomes and other DNA-associated proteins. SWI2/SNF2 proteins contain sequence motifs characteristic of SF2 helicases but do not have helicase activity. Instead, they couple ATP hydrolysis with the generation of superhelical torsion in DNA. The structure of the nucleosome-remodeling domain of zebrafish Rad54, a protein involved in Rad51-mediated homologous recombination, reveals that the core of the SWI2/SNF2 enzymes consist of two alpha/beta-lobes similar to SF2 helicases. The Rad54 helicase lobes contain insertions that form two helical domains, one within each lobe. These insertions contain SWI2/SNF2-specific sequence motifs likely to be central to SWI2/SNF2 function. A broad cleft formed by the two lobes and flanked by the helical insertions contains residues conserved in SWI2/SNF2 proteins and motifs implicated in DNA-binding by SF2 helicases. The Rad54 structure suggests that SWI2/SNF2 proteins use a mechanism analogous to helicases to translocate on dsDNA. PMID- 15806109 TI - Effect of mast cell chymase inhibitor on the development of scleroderma in tight skin mice. AB - 1 Although the pathogenesis of scleroderma is not fully understood, activation of connective-tissue-type mast cells (CTMCs) has been implicated in various fibrotic diseases. 2 Our previous study showed that the number of CTMCs was markedly increased during fibrous proliferation in the skin of a scleroderma model, namely tight-skin (Tsk) mice. Because mast cells express numerous bioactive factors, such as cytokines, growth factors, proteases, and others, it is crucial to identify the primary factors that may be involved in the pathogenesis of scleroderma. Our previous study also showed that a CTMC-specific protease, chymase-4, was selectively upregulated in accordance with the development of skin fibrosis in Tsk mice. 3 To further elucidate the role of chymase secreted from CTMCs, we evaluated the therapeutic effects of a synthetic chymase-specific inhibitor, SUN-C8257, on the development of skin fibrosis in Tsk mice. SUN-C8257 (50 mg kg-1 day-1) was administered via intraperitoneal injection in 13-week-old Tsk mice for a period of 2 weeks. 4 Treatment with SUN-C8257 significantly reduced chymase activity by 43% and the chymase-4 mRNA level by 47%, and also decreased the thickness of the subcutaneous fibrous layer of Tsk mice by 42% compared with that of Tsk mice injected with vehicle. 5 Furthermore, immunohistochemical analysis revealed that transforming growth factor (TGF)-beta1 staining in the fibrous layer of Tsk skin was markedly reduced by the treatment with SUN-C8257. This chymase inhibitor may prevent the chymase-dependent pathway that activates the latent TGF-beta1 in fibrous tissue, and may exhibit beneficial effects that inhibit the development of fibrosis. 6 In conclusion, our results strongly support the assumption that CTMC-derived chymase may play a key role in the pathogenesis of scleroderma. PMID- 15806110 TI - Mechanisms of the protective effects of urocortin on coronary endothelial function during ischemia-reperfusion in rat isolated hearts. AB - 1 Urocortin is a vasodilator peptide related to corticotrophin-releasing factor, which may protect endothelial function during coronary ischemia-reperfusion (I R). The aim of this study was to study the mechanisms of this protective effect. 2 Hearts from Sprague-Dawley rats were isolated and perfused at constant flow and then exposed to 15 min global zero-flow ischemia, followed by 15 min reperfusion. The relaxation to acetylcholine (10 nM-10 microM) was recorded after pre constriction of the coronary vasculature with U46619 (100-300 nM) in ischemic reperfused or time-control hearts. 3 After I-R, the coronary relaxation to acetylcholine was reduced and this reduction was attenuated by treatment with urocortin (10 pM), administered before ischemia and during reperfusion. 4 This urocortin-induced improvement of the relaxation to acetylcholine was not modified by tetraethylammonium (10 mM), blocker of Ca2+ dependent-potassium channels; glibenclamide (10 microM), blocker of K(ATP) channels; N(w)-nitro-L-arginine methyl ester (L-NAME, 100 microM), blocker of nitric oxide synthesis; or meclofenamate (10 microM), blocker of cyclooxygenase, but it was abolished by chelerythrine (3 microM), blocker of protein kinase C (PKC). 5 These results suggest that urocortin may protect coronary endothelial function during I-R by activation of PKC. PMID- 15806111 TI - Protein kinase C-alpha mediates TNF release process in RBL-2H3 mast cells. AB - 1 To clarify the mechanism of mast cell TNF secretion, especially its release process after being produced, we utilized an antiallergic drug, azelastine (4-(p chlorobenzyl)-2-(hexahydro-1-methyl-1H-azepin-4-yl)-1-(2H)- phthalazinone), which has been reported to inhibit TNF release without affecting its production in ionomycin-stimulated RBL-2H3 cells. 2 Such inhibition was associated with the suppression of an ionomycin-induced increase in membrane-associated PKC activity rather than the suppression of Ca2+ influx, suggesting that PKC might be involved in TNF release process. 3 To see whether conventional PKC family (cPKCs) are involved, we investigated the effects of a selective cPKC inhibitor (Go6976) and an activator (thymeleatoxin) on TNF release by adding them 1 h after cell stimulation. By this time, TNF mRNA expression had reached its maximum. Go6976 markedly inhibited TNF release, whereas thymeleatoxin enhanced it, showing a key role of cPKC in TNF post-transcriptional process, possibly its releasing step. 4 To determine which subtype of cPKCs could be affected by azelastine, Western blotting and live imaging by confocal microscopy were conducted to detect the translocation of endogenous cPKC (alpha, betaI and betaII) and transfected GFP tagged cPKC, respectively. Both methods clearly demonstrated that 1 microM azelastine selectively inhibits ionomycin-triggered translocation of (alpha)PKC without acting on betaI or betaIIPKC. 5 In antigen-stimulated cells, such a low concentration of azelastine did not affect either (alpha)PKC translocation or TNF release, suggesting a functional link between (alpha)PKC and the TNF-releasing step. 6 These results suggest that (alpha)PKC mediates the TNF release process and azelastine inhibits TNF release by selectively interfering with the recruitment of (alpha)PKC in the pathway activated by ionomycin in RBL-2H3 cells. PMID- 15806112 TI - Ciclopirox protects mitochondria from hydrogen peroxide toxicity. AB - 1 The mitochondrial respiratory chain produces reactive oxygen species (ROS) during normal electron transport. Despite producing ROS, mitochondria are vulnerable to oxidative stress. Mitochondrial dysfunction has been associated with many degenerative diseases, making it important to identify compounds that protect mitochondria from ROS-mediated toxicity. Here we report that ciclopirox (CPX) blocks H2O2-induced mitochondrial injury by maintaining mitochondrial transmembrane potential (Deltapsim). 2 CPX completely blocked H2O2-stimulated release of lactate dehydrogenase (a marker of cell death) and decrease in MTT reduction (a marker of mitochondrial function) in adenocarcinoma SK-HEP-1 cells. 3 H2O2 rapidly depolarized the Deltapsim, and CPX blocked this H2O2-stimulated Deltapsim decrease. Similar data were obtained in experiments using mitochondria isolated from rat liver. 4 Furthermore, CPX effectively inhibited H2O2-induced mitochondrial permeability transition pore (MPTP) opening. In de-energized mitochondria, however, CPX did not inhibit Ca2+-evoked MPTP opening, indicating that CPX is not a direct inhibitor of the MPTP. 5 Oxygen consumption studies showed that in the presence of pyruvate and malate CPX restored the rate of state 3 to state 4 respiration decreased by H2O2. Consistent with this, CPX replenished ATP levels lowered by H2O2. 6 The present results indicate that CPX protects SK HEP-1 cells from H2O2 cytotoxicity by inhibiting Deltapsim decrease and indirectly preventing MPTP opening. PMID- 15806113 TI - Artocarpol A stimulation of superoxide anion generation in neutrophils involved the activation of PLC, PKC and p38 mitogen-activated PK signaling pathways. AB - 1 Artocarpol A (ART), a natural phenolic compound isolated from Artocarpus rigida, stimulated a slow onset and long-lasting superoxide anion generation in rat neutrophils, whereas only slightly activated the NADPH oxidase in a cell-free system. 2 Pretreatment of neutrophils with pertussis toxin (1 microg ml(-1)), 50 microM 2'-amino-3'-methoxyflavone (PD 98059), or 1 microM 1,4-diamino-2,3-dicyano 1,4-bis(2-aminophenylthio)butadiene (U0126) had no effect on ART-stimulated superoxide anion generation. ART (30 microM) did not induce extracellular signal regulated kinase (ERK) phosphorylation. 3 4-(4-Fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole (SB 203580) markedly attenuated the ART-stimulated superoxide anion generation (IC50 value of 4.3+/-0.3 microM). Moreover, ART induced p38 mitogen-activated PK (MAPK) phosphorylation and activation. 4 The superoxide anion generation in response to ART was also substantially inhibited in a Ca2+-free medium, and by pretreatment with 1 microM 1-[6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-1H-pyrrole-2,5 dione (U-73122) and 100 microM 2-aminoethyldiphenyl borate (2-APB). ART (30 microM) stimulated the [Ca2+]i elevation in the presence or absence of external Ca2+, and also increased the D-myo-inositol 1,4,5-trisphosphate formation. 5 2-[1 (3-Dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimide (GF 109203X) greatly inhibited the ART-stimulated superoxide anion generation (IC50 value of 7.8+/-1.0 nM). ART increased the recruitment of PKC-alpha, -betaI, and -betaII to the plasma membrane of neutrophils, and stimulated Ca2+-dependent PKC activation in the cytosol preparation. 6 ART induced the phosphorylation of p47phox, which was attenuated by GF 109203X. Moreover, ART evoked the membrane association of p47(phox), which was inhibited by GF 109203X and SB 203580. 7 These results indicate that the ART stimulation of superoxide anion generation involved the activation of p38 MAPK, PLC/Ca2+, and PKC signaling pathways in rat neutrophils. PMID- 15806114 TI - D-aspartate and NMDA, but not L-aspartate, block AMPA receptors in rat hippocampal neurons. AB - 1 The amino acid, D-aspartate, exists in the mammalian brain and is an agonist at the N-methyl-D-aspartate (NMDA) subtype of ionotropic glutamate receptors. Here, for the first time, we studied the actions of D-aspartate on alpha-amino-3 hydroxyl-5-methyl-4-isoxazolepropionate receptors (AMPARs) in acutely isolated rat hippocampal neurons. 2 In the presence of the NMDA receptor channel blocker, MK801, D-aspartate inhibited kainate-induced AMPAR current in hippocampal neurons. The inhibitory action of D-aspartate on kainate-induced AMPAR current was concentration-dependent and was voltage-independent in the tested voltage range (-80 to +60 mV). 3 The estimated EC50 of the L-glutamate-induced AMPAR current was increased in the presence of D-aspartate, while the estimated maximum L-glutamate-induced AMPAR current was not changed. D-aspartate concentration dependently shifted the dose-response curve of kainate to the right. Schild plot analysis indicated that D-aspartate acts competitively to block AMPARs. The K(b) for D-aspartate was estimated to be 0.93 mM. 4 D-aspartate also blocked L glutamate-induced current in Xenopus laevis oocytes that expressed recombinant homomeric AMPARs. 5 NMDA possessed similar inhibitory action on AMPARs. However, L-aspartate had little inhibitory action on AMPARs. 6 D-Aspartate, but not L aspartate, was found to reduce the amplitude of miniature excitatory postsynaptic current in cultured hippocampal neurons. 7 Our data are consistent with a model in which D-aspartate directly competes with kainate and L-glutamate in binding to the agonist binding site of AMPARs. The prevalence of D-aspartate in the brain suggests a possible role of D-aspartate in modulating AMPAR-mediated fast excitatory synaptic transmission. PMID- 15806115 TI - Block of TRPC5 channels by 2-aminoethoxydiphenyl borate: a differential, extracellular and voltage-dependent effect. AB - 1 2-aminoethoxydiphenyl borate (2-APB) has been widely used to examine the roles of inositol 1,4,5-trisphosphate receptors (IP3Rs) and store-operated Ca2+ entry and is an emerging modulator of cationic channels encoded by transient receptor potential (TRP) genes. 2 Using Ca2+-indicator dye and patch-clamp recording we first examined the blocking effect of 2-APB on human TRPC5 channels expressed in HEK-293 cells. 3 The concentration-response curve has an IC50 of 20 microM and slope close to 1.0, suggesting one 2-APB molecule binds per channel. The blocking effect is not shared by other Ca2+ channel blockers including methoxyverapamil, nifedipine, N-propargylnitrendipine, or berberine. 4 In whole-cell and excised membrane patch recordings, 2-APB acts from the extracellular but not intracellular face of the membrane. 5 Block of TRPC5 by 2-APB is less at positive voltages, suggesting that it enters the electric field or acts by modulating channel gating. 6 2-APB also blocks TRPC6 and TRPM3 expressed in HEK-293 cells, but not TRPM2. 7 Block of TRP channels by 2-APB may be relevant to cell proliferation because 2-APB has a greater inhibitory effect on proliferation in cells overexpressing TRPC5. 8 Our data indicate a specific and functionally important binding site on TRPC5 that enables block by 2-APB. The site is only available via an extracellular route and the block shows mild voltage-dependence. PMID- 15806116 TI - Glutamate excitotoxicity in glaucoma: truth or fiction? PMID- 15806118 TI - Optimal initial dose of oral cyclosporine in relation to its toxicities for graft versus-host disease prophylaxis following reduced-intensity stem cell transplantation in Japanese patients. AB - Since the introduction of reduced-intensity stem-cell transplantation (RIST), allogeneic stem-cell transplantation has become available for elderly patients. While pharmacokinetics of cyclosporine might differ according to age or other factors, cyclosporine is uniformly started at an oral dose of 6 mg/kg/day. We retrospectively reviewed medical records of 35 patients aged between 32 and 65 (median 52) years who had undergone RIST. Doses of cyclosporine were adjusted to the target blood trough level of 150-250 ng/ml. Cyclosporine dosages were changed in 33 patients (94%). Dose reduction was required in 32 patients because of high blood levels (n=25), renal dysfunction (n=3), hepatic dysfunction (n=2), and hypertension (n=2). Cyclosporine doses were increased in one because of the suboptimal level. The median of the achieved stable doses was 3.1 mg/kg/day (range, 1.0-7.4). Five patients sustained Grade III toxicities according to NCI CTC version 2.0: renal dysfunction (n=4), hyperbilirubinemia (n=2), and hypertension (n=2). No patients developed grade IV toxicity. There was no statistically significant difference in the frequency and severity of cyclosporine toxicities between patients aged 50 years and above and those below 50 years. The initial oral cyclosporine dose of 6 mg/kg/day was unnecessarily high irrespective of age. The possible overdose of cyclosporine might have aggravated regimen-related toxicities. PMID- 15806119 TI - The effect of low-dose aciclovir on reactivation of varicella zoster virus after allogeneic haemopoietic stem cell transplantation. AB - Patients undergoing haemopoietic stem cell transplants (HSCT) are at high risk of varicella zoster virus (VZV) reactivation, with a significant incidence of dissemination. This study reports a retrospective analysis of 247 allogeneic HSCT recipients receiving anti-viral prophylaxis with low-dose oral aciclovir 400 mg/day, administered until immunosuppression was discontinued and the CD4(+) cell count exceeded 200/mm(3). Viral reactivation was successfully suppressed by aciclovir prophylaxis, with only one case of breakthrough infection. The cumulative incidence of zoster infection at 1 year post transplant was 2% and at 5 years 34%. In all, 64 patients discontinued prophylaxis. Zoster developed in 26 of these, giving a cumulative incidence of infection at 1 year after stopping aciclovir of 39% and at 3 years 44%. Infection occurred in a localised dermatomal distribution in 93% of cases. This supports previous findings that aciclovir prophylaxis prevents early VZV reactivation, although the long-term incidence is not affected as infection occurs once prophylaxis is discontinued. Such infection, however, is mild and localised. This study does not support the idea that use of such low-dose aciclovir regimens reduces the zoster incidence by permitting subclinical reactivation during prophylaxis, and therefore the re establishment of protective anti-viral immunity. PMID- 15806120 TI - Factors affecting the outcome of stem cell transplantation from unrelated donors for childhood acute lymphoblastic leukemia in third remission. AB - Between July 1990 and March 2002, 35 consecutive children with ALL in third complete remission (CR3) underwent stem cell transplantation (SCT) from unrelated donors (UD). All patients received CAMPATH-1M 5-20 mg daily for 5 days. Grafts were T-cell depleted in 30 patients, 29 by CAMPATH antibodies and one by CD34 selection. Median follow-up was 3.8 years (0.3-9.3). Event-free survival (EFS) at 3 years was 35% (SE 8%); relapse rate and transplant-related mortality (TRM) at 3 years was 42 and 23%. Short first complete remission (CR1) <2.5 years was associated with lower EFS (P=0.001), higher TRM (P=0.019) and higher relapse rate (P=0.023). Short second complete remission (CR2) <2.5 years was associated with lower EFS (P=0.003) and higher TRM (0.009). Higher relapse rate and lower EFS were associated with isolated first extramedullary relapse (P=0.019, 0.012). There was no significant difference in outcome between mismatched unrelated donor stem cell transplantation (MMUD-SCT) and matched unrelated donor stem cell transplantation (UD-SCT). We conclude that UD-SCT is an effective treatment of ALL in CR3. The outcome remains limited by TRM and a high relapse rate. Short duration of CR1 and of CR2 and extramedullary site at first relapse are particularly adverse. MMUD should also be considered in high-risk patients, since the outcome of MMUD appears similar to MUD. PMID- 15806121 TI - T-cell reconstitution and expansion after hematopoietic stem cell transplantation: 'T' it up! AB - Adoptive immunotherapy is the isolation and infusion of antigen-specific or nonspecific lymphocytes. Adoptive therapy with T cells may have a role in replacing, repairing, or enhancing immune function damaged by cytotoxic therapies, and rapid lymphocyte recovery may improve outcome after autologous and allogeneic stem cell transplantation (SCT). Recently, a plethora of information on the basic mechanisms of T-cell biology and regulation of cellular immune responses has emerged, permitting the development of new forms of adoptive cell therapy. Efficient ex vivo culture method for T-cell subsets affords the possibility of adoptive transfer of T cells engineered with enhanced capacity for central memory, effector cytotoxicity, Th1, Th2, veto cell, and T regulatory functions. Studies show that homeostatic T-cell proliferation is important for effective adoptive immunotherapy and pretreatment with chemotherapy may enhance the effects of infused T cells. Replicative senescence, in part due to telomere erosion, likely limits successful adoptive immunotherapy, though it may be possible to maintain T-cell pools by enforced expression of telomerase. Clinical trials now demonstrate that it is possible to enhance immune reconstitution after SCT with cytokines or infusions of ex vivo costimulated expanded T cells. These data all support the premise that adoptive therapy can accelerate reconstitution of cellular immunity with enhanced antitumor effects following SCT. PMID- 15806123 TI - Ancestim associated with Filgrastim and/or chemotherapy can improve blood progenitor yields in patients who previously failed mobilisation. PMID- 15806124 TI - Successful non-T-cell-depleted HLA-haploidentical stem cell transplantation (SCT) with reduced-intensity conditioning from a second child for late graft failure after the first HLA-haploidentical SCT for MDS/overt leukemia based on feto maternal microchimerism. PMID- 15806125 TI - Melphalan 200 mg/m2 with blood stem cell support as first-line myeloma therapy: impact of glomerular filtration rate on engraftment, transplantation-related toxicity and survival. AB - In this retrospective single-centre study, 96 consecutive myeloma patients were treated with melphalan 200 mg/m(2) with blood stem cell support as first-line therapy. Their mean age was 55 (38-65) years. The impact of renal function on stem cell collection yield, engraftment, transplantation-related toxicity and overall survival was studied. Glomerular filtration rate (GFR) was evaluated by iohexol clearance, a median 32 days before high-dose administration. Chronic renal failure (GFR <60 ml/min) was present in 19 patients, with severe failure (GFR <30 ml/min) in five patients, including one patient on haemodialysis. No relationship between GFR and stem cell collection yield or engraftment was observed, nor was the incidence of neutropenic fever or infectious complications related to GFR. Patients with subnormal renal function, however, were more often affected by severe mucositis. In addition, the two patients with severe GI bleeding, the two pneumonia patients who needed ventilator support and the only therapy-related death were noted in the five patients with severe renal failure. Lower iohexol clearance at the time of high-dose administration was found to have a poor impact on survival. A reduction of melphalan dose in patients with severe renal failure, here defined as iohexol clearance <30 ml/min, is suggested. PMID- 15806126 TI - Late thyroid toxicity in 153 long-term survivors of allogeneic bone marrow transplantation for acute lymphoblastic leukaemia. AB - The purpose of this study was to identify risk factors for hypothyroidism after bone marrow transplantation (BMT) for high-risk or relapsed acute lymphoblastic leukaemia (ALL) in children. In all, 388 children with acute lymphoblastic leukaemia underwent allogeneic bone marrow transplantation between 1984 and 1994. Overall 5-year survival was 54.6%. Thyroid function was assessed in the 153 patients with more than 5 years of follow-up. In total, 16 patients developed uncompensated hypothyroidism (UH) and 46 compensated hypothyroidism (CH) a median of 2.9 and 2.7 years, respectively, after BMT. Thyroid dysfunction-free survival rates were 73.2% after 5 years and 59.2% after 10 years. Three factors were significantly associated with the onset of hypothyroidism, namely age, bone marrow transplantation in second remission, and single-dose total body irradiation (TBI). Ultrasonography of the thyroid showed nodules in 10 of 35 patients. The median time from BMT to nodule detection was 7.8 years. Cytology (n=5) and surgery (n=4) showed no evidence of thyroid cancer. Four of the 14 patients who received cytoreduction without TBI but with busulphan and cyclophosphamide developed UH (n=2) or CH (n=2). We concluded that children who undergo BMT for ALL are at a high risk of subsequent thyroid dysfunction. PMID- 15806127 TI - Polyoma BK viruria in patients undergoing autologous hematopoietic stem cell transplantation. PMID- 15806128 TI - Reduced-intensity allogeneic stem cell transplantation in relapsed and refractory Hodgkin's disease: low transplant-related mortality and impact of intensity of conditioning regimen. AB - A total of 40 patients with relapsed/refractory Hodgkin's disease (HD) underwent reduced-intensity conditioning (RIC) allogeneic stem cell transplantation (allo SCT) from an HLA-identical sibling (n=20) or a matched unrelated donor (n=20). The median age was 31 years (range 18-58). Disease status at allo-SCT was refractory relapse (n=14) or sensitive relapse (n=26). The conditioning regimens were fludarabine-cyclophosphamide+/-antithymocyte globulin (n=14), a less intensive regimen, and fludarabine-melphalan (FM) (n=26), a more intensive one. The two groups had similar prognostic factors. The median time to neutrophil recovery (ie absolute neutrophil count >/=500/microl) was 12 days (range 10-24). The median time to platelet recovery (ie platelet count >/=20 000/microl) was 17 days (range 7-132). Day 100 and cumulative (18-month) transplant-related mortalities (TRMs) were 5 and 22%. Twenty-four patients (60%) are alive (14 in complete remission or complete remission, unconfirmed/uncertain) with a median follow-up of 13 months (4-78). In all, 16 patients expired (TRM n=8, disease progression n=8). FM patients had better overall survival (73 vs 39% at 18 months; P=0.03), and a trend towards better progression-free survival (37 vs 21% at 18 months; P=0.2). RIC allo-SCT is feasible in relapsed/refractory HD patients with a low TRM. The intensity of the preparative regimen affects survival. PMID- 15806129 TI - VDRL screening and haemopoietic stem cell transplantation. PMID- 15806130 TI - In vivo T-cell depletion with low-dose ATG is effective in reducing cGVHD after peripheral blood stem cell myeloablative sibling transplants in CML: results from a prospective phase II study. PMID- 15806131 TI - Impact of disease burden at time of allogeneic stem cell transplantation in adults with acute myeloid leukemia and myelodysplastic syndromes. AB - The impact of disease burden on the outcome of patients with acute myeloid leukemia (AML) undergoing allogeneic stem cell transplantation (SCT) has not been well defined. Data from several retrospective series suggest that overt leukemia at the time of transplant increases the risk of relapse. We reviewed the outcomes of 68 consecutive adults with AML (n=60) or myelodysplastic syndromes (MDS) (n=8) who received an allogeneic SCT at the University of Chicago between May 1986 and October 2002 to confirm the importance of currently recognized risk factors for overall survival (OS) and progression-free survival (PFS). In addition, we wanted to determine whether quantification of residual disease by blast percentage or cytogenetic abnormalities at the time of SCT was correlated with outcome. AML subtypes based on the FAB classification were as follows: M0=9, M1=9, M2=16, M3=2, M4=16, M5=3, M6=5. Cytogenetic analysis was available from 52 patients. Using standard morphologic criteria, 34 patients were in complete remission (CR) and 34 had visible leukemia present. The majority of donors were HLA-identical siblings (n=55). In all, 56 patients received myeloablative conditioning regimens and 12 received a reduced-intensity, fludarabine-based conditioning regimen. OS and PFS times were 7.1 months (95% CI, 4.8-10.4) and 5.1 months (95% CI, 3.2 7.8), respectively. Median follow-up from SCT was 4.6 years (range, 0.6-17.0) for survivors. In multivariate analysis, the following factors were found to be associated with worse survival: (1) increased percentage of blasts in the bone marrow at the time of SCT, (2) presence of acute graft-versus-host disease, (3) mismatched donor, (4) Zubrod performance score of >/=2, and (5) age >/=45 years. We also found a trend towards improved outcome among patients in cytogenetic remission as compared to those who had residual cytogenetic abnormalities and those in overt relapse. These data support an association between pre-transplant disease burden and poor outcome after SCT. PMID- 15806132 TI - Pilot study of 13cis-retinoic acid+dexamethasone+alpha interferon as maintenance therapy following high-dose chemotherapy and autologous stem cell transplant for multiple myeloma. AB - Interleukin 6 (IL-6) is a major growth factor for myeloma cells and retinoids have been shown to inhibit expression of the interleukin 6 receptor (IL-6R). We performed a pilot study to assess the efficacy and tolerability of 13cis retinoic acid (13cRA) and dexamethasone (Dex), when added to interferon alpha (IFNalpha) as maintenance therapy post autologous stem cell transplantation. Between 90 and 120 days post stem cell transplantation, 33 patients were started on 13cRA 1 mg/kg p.o. daily for 14 days and Dex 40 mg p.o daily for 5 days every month. 13cRA was dose escalated by 0.5 mg/kg/month to 2 mg/kg. Seventeen patients had a persistent paraprotein post transplant. Overall, a response to therapy was observed in 11/17 (64%), with a complete response in 4/17 (23.5%) and a partial response (>/=50% paraprotein decline) in 7/17 (41%). With a median follow-up of 34.8 months, 22/33 (66%) demonstrated disease progression and 11/33 (33%) died. The median progression-free survival from diagnosis was 34.7 months. Although a decline in paraprotein was frequently observed on triple therapy, many patients discontinued therapy due to the side-effects of the IFNalpha. Future trials should be designed using 13cRA and Dex alone. PMID- 15806133 TI - Nonmyeloablative stem cell transplantation using lymphoablative rather than myeloablative conditioning in the prefludarabine era by ATG and limiting doses of cyclophosphamide. AB - The feasibility of using lymphoablative rather than myeloablative conditioning for durable engraftment of allogeneic stem cells and subsequent cell therapy with donor lymphocytes was pioneered in the prefludarabine era in patients with resistant lymphoma and metastatic solid tumors. Between July 1995 and August 1996, 15 patients, five males and 10 females, median age 50 (range 20-57) years, were enrolled in a protocol that consisted of different doses of cyclophosphamide (Cy), 50 mg/kg/day for 1, 2, 3 or 4 consecutive days in parallel with a fixed dose of rabbit antithymocyte globulin (ATG) (Fresenius) 10 mg/kg/day for 4 consecutive days. All patients, except one treated with a single dose of Cy, achieved full tri-lineage engraftment and no late graft failure was observed. Only three patients suffered from grade III-IV graft-versus-host disease (GVHD). Three patients out of the 15 survived long term (follow-up >93 to >96 months). We concluded that lymphoablative conditioning with ATG and intermediate-to-high-dose Cy is well tolerated and can result in durable engraftment with acceptable GVHD in heavily pretreated patients with advanced malignancies. Hence, induction of tolerance to donor alloantigens by lymphoablative conditioning while avoiding myeloablative chemotherapy or radiation therapy may serve as a platform for subsequent cell therapy with donor lymphocytes. PMID- 15806134 TI - Failure of anti-infective mouth rinses and concomitant antibiotic prophylaxis to decrease oral mucosal colonization in autologous stem cell transplantation. AB - Autologous stem cell transplantation has augmented treatment successes. However, high-dose chemotherapy is still accompanied by dose-limiting toxicities, for example, severe mucositis. Mucosal lesions serve as portals of entry for infections. In order to reduce the oral microbial burden, we prospectively evaluated the microbiological impact of a complex regimen of mouth rinses consisting of concomitantly applied polyene antifungals, povidone-iodine, chlorhexidine, sage tea, and prophylactic ciprofloxacin and fluconazole. A total of 15 patients were enrolled into this longitudinal evaluation. Colony-forming units (CFU) were quantitated from saliva, buccal and palatinal swabs during high dose chemotherapy and autologous stem cell transplantation. The number of CFU did not show any significant changes after initiation of the mouth rinses and the prophylactic antibiotics. The median CFU count was 268 x 10(6)/ml saliva before chemotherapy and decreased after initiation of intravenous antibiotics only. Neither prophylactic nor therapeutic antifungals significantly reduced the number of cultures positive for yeasts. Since 90% of our patients had febrile neutropenia at some time point during the observation period, the approach evaluated cannot be recommended as prophylaxis of febrile neutropenia as such. PMID- 15806135 TI - Treatment of steroid-resistant acute graft-versus-host disease with daclizumab and etanercept. AB - Steroid-resistant acute GVHD (aGVHD) following allogeneic hematopoietic stem cell transplantation (alloHSCT) continues to be associated with a high mortality. We report the results of a phase II study of treatment of steroid-resistant aGVHD with the IL-2 receptor antibody daclizumab combined with the TNF-receptor fusion protein etanercept. Treatment consisted of daclizumab 1 mg/kg given i.v. on days 1, 4, 8, 15, 22 and etanercept 16 mg/m(2) s.c. on days 1, 5, 9, 13, 17. A total of 21 patients (age 15-61 years) with steroid-resistant aGVHD after alloHSCT were included in the study. Donor types were HLA-matched related (n=6), HLA-matched unrelated (n=14), and HLA-mismatched unrelated (n=1). Eight patients achieved complete, and six showed partial remission of aGVHD. Seven patients did not respond. Four of 21 patients are currently alive with a median follow-up of 586 (185-1155) days. Three patients died due to relapsed malignancy. Treatment related mortality was due to infectious complications (n=11) or organ failure due to aGVHD (n=3). In total, 12 patients developed subsequent chronic GVHD. In conclusion, the data demonstrate an acceptable response rate of the combination of daclizumab and etanercept in the treatment of steroid-resistant aGVHD. Nevertheless, long-term mortality due to infectious complications and chronic GVHD remains high. PMID- 15806136 TI - ADAMTS13 is expressed in hepatic stellate cells. AB - ADAMTS13 is a circulating zinc metalloprotease that cleaves the hemostatic glycoprotein von Willebrand factor (VWF) in a shear-dependent manner. Deficiency in ADAMTS13, owing to genetic mutations or autoimmune inhibitors, causes thrombotic thrombocytopenic purpura (TPP). Northern blot analysis has shown that ADAMTS13 is expressed primarily in the liver. By using real-time RT-PCR, we confirmed that in mice the liver had the highest level of the ADAMTS13 transcript. To identify the liver cell-type-specific origin of ADAMTS13, we used in situ hybridization techniques to investigate the pattern of ADAMTS13 expression in the liver; analyzed the ADAMTS13 proteolytic activity in the culture media of fractionated liver cells; and confirmed ADAMTS13 expression with RT-PCR analysis and cloning of the mouse ADAMTS13 gene. The results revealed that ADAMTS13 was expressed primarily in cell fractions enriched in hepatic stellate cells. The mouse ADAMTS13 cloned from primary hepatic stellate cells was similar to its human counterpart in digesting VWF and was susceptible to suppression by EDTA or the IgG inhibitors of patients with TTP. Since hepatic stellate cells are believed to play a major role in the development of hepatic fibrosis and cirrhosis, the identification of the liver cell-type expressing ADAMTS13 will have important implications for understanding pathophysiological mechanisms regulating ADAMTS13 expression. PMID- 15806137 TI - Characterization of lymphoblast mitochondria from patients with Barth syndrome. AB - Barth syndrome (BTHS) is a multisystem disorder of individuals who carry mutations in tafazzin, a putative phospholipid acyltransferase. We investigated the hypothesis that BTHS is caused by specific impairment of the mitochondrial lipid metabolism. The fatty acid composition of all major mitochondrial phospholipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE), and cardiolipin (CL), changed in lymphoblasts from BTHS patients. These changes were most extensive in CL and least extensive in PE. The complementary nature of the fatty acid alterations in CL and PC suggested that fatty acid transfer between these two lipids was inhibited in BTHS. Fluorescence staining and electron microscopy showed abnormal proliferation of mitochondria in BTHS lymphoblasts. The mitochondrial membrane potential, monitored with the fluorescence probe JC-1, was reduced in BTHS lymphoblasts. However, mitochondrial ATP formation of permeabilized lymphoblasts remained unaffected in BTHS. The data suggest that phospholipid abnormalities of BTHS mitochondria led to partial uncoupling of oxidative phosphorylation and that lymphoblasts compensated for this deficiency by expanding the mitochondrial compartment. PMID- 15806138 TI - Upregulation of TCF4 expression as a transcriptional target of beta-catenin/p300 complexes during trans-differentiation of endometrial carcinoma cells. AB - Nuclear stabilization of beta-catenin and its interaction with TCF/LEF factors are key events in transduction of the Wnt/beta-catenin signal pathway. Our previous study indicated that nuclear beta-catenin accumulation provides an initial signal for trans-differentiation toward the squamoid phenotype of endometrial carcinoma (Em Ca) cells in a TCF4-dependent manner, which makes this a possible factor for a positive prognosis. However, little is known about regulation of TCF4 expression in Em Cas. We show here that beta-catenin can directly induce transcription from the TCF4 promoter, the effect being enhanced by the p300 coactivator. In clinical cases, nuclear beta-catenin accumulation was found to frequently overlap with TCF4 immunoreactivity in morules and surrounding glandular carcinoma lesions, showing a significant positive correlation (r = 0.82, P < 0.0001), in contrast to areas of squamous metaplasia (SqM) within Em Cas. In cases with coexistence of two squamoid features in trans-differentiated areas, loss of nuclear beta-catenin and TCF4 immunoreactivity was closely related to change in the morphology from the morular to the SqM phenotype. The TCF4 promoter contains a single consensus TCF-binding site that is critical for activation by beta-catenin. The p300 coactivator, in particular N-terminal residues 1 to 670, appears sufficient to enhance beta-catenin-dependent transcription, again with TCF4-dependence. These findings indicate that a positive feedback loop of TCF4 expression mediated by beta-catenin/p300 may be important for initial steps during trans-differentiation of Em Ca cells. In addition, its downregulation is associated with induction of a more differentiated squamoid phenotype. PMID- 15806139 TI - Expression of HSP47, a collagen-specific chaperone, in normal and diseased human liver. AB - HSP47 is a collagen-specific chaperone that is required for normal collagen synthesis. In animal models of liver injury, hepatic stellate cells (HSC) have been identified as a source of HSP47. Because expression of HSP47 has not been investigated in human liver, the aim of these studies was to characterize expression of HSP47 in human liver and to investigate its regulation in human HSC in vitro. Immunohistochemistry demonstrated staining for HSP47 along the sinusoids of normal and cirrhotic human livers and in fibrous septa. Dual fluorescence confocal microscopy showed colocalization of HSP47 with synaptophysin, a marker for HSC. Levels of immunoreactive HSP47 and its transcript tended to be higher in cirrhotic livers than in normal livers. The abundance of HSP47 protein was unchanged by treatment of cultured human HSC with TGF-beta1, angiotensin II, hypoxia and a number of other treatments intended to increase collagen synthesis. A modest reduction in HSP47 was achieved by transfection with antisense oligonucleotides and was associated with a significant decrease in procollagen synthesis. These observations suggest that HSP47 is constitutively expressed in human HSC and that HSP47 may be a target for antifibrotic therapy. PMID- 15806140 TI - Vascular leakage in chick embryos after expression of a secreted binding protein for fibroblast growth factors. AB - Fibroblast growth factors (FGFs) have been implicated in a variety of physiologic and pathologic processes from embryonic development to tumor growth and angiogenesis. FGFs are immobilized in the extracellular matrix of different tissues and require release from this storage site to trigger a response. Secreted FGF-binding proteins (FGF-BPs) can release immobilized FGFs, enhance the activity of locally stored FGFs and can thus serve as an angiogenic switch molecule in cancer. Here, we report on the effect of human FGF-BP transgene expression in chicken embryos. To establish the transgenic model, plasmid-based reporter vectors expressing luciferase, beta-galactosidase or green fluorescent protein were introduced through different routes into 4- to 5-day-old embryos grown outside their egg shell on top of the yolk sac. This allows for easy manipulation and continuous observation of phenotypic effects. Expression of human FGF-BP induced dose-dependent vascular permeability, hemorrhage and embryonic lethality. Light and electron microscopic studies indicate that this hemorrhage results from compromised microvascular structure. An FGF-1 expression vector with an added secretory signal mimicked this vascular leakiness phenotype whereas wild-type FGF-1 required coexpression of a threshold amount of FGF-BP. This model is a powerful tool for real-time monitoring of the effects of transient transgene expression during embryogenesis. PMID- 15806141 TI - c-FMS chromatin structure and expression in normal and leukaemic myelopoiesis. AB - The macrophage colony-stimulating factor receptor is encoded by the c-FMS gene, and it has been suggested that altered regulation of c-FMS expression may contribute to leukaemic transformation. c-FMS is expressed in pluripotent haemopoietic precursor cells and is subsequently upregulated during monocytic differentiation, but downregulated during granulopoiesis. We have examined transcription factor occupancy and aspects of chromatin structure of the critical c-FMS regulatory element located within the second intron (FIRE - fms intonic regulatory element) during normal and leukaemic myelopoiesis. Granulocytic differentiation from normal and leukaemic precursors is accompanied by loss of transcription factors at FIRE and downregulated c-FMS expression. The presence of AML1-ETO in leukaemic cells does not prevent this disassembly. In nonleukaemic cells, granulocytic differentiation is accompanied by reversal to a chromatin fine structure characteristic of c-FMS-nonexpressing cells. In addition, we show that low-level expression of the gene in leukaemic blast cells and granulocytes does not associate with increased CpG methylation across the c-FMS locus. PMID- 15806142 TI - Significance of HDAC6 regulation via estrogen signaling for cell motility and prognosis in estrogen receptor-positive breast cancer. AB - Histone deacetylase (HDAC) 6 is a subtype of the HDAC family; it deacetylates alpha-tubulin and increases cell motility. Here, we investigate the impact of an alteration of HDAC6 expression in estrogen receptor alpha (ER)-positive breast cancer MCF-7 cells, as we identified that HDAC6 is a novel estrogen-regulated gene. MCF-7 treated with estradiol showed increased expression of HDAC6 mRNA and protein and a four-fold increase in cell motility in a migration assay. Cell motility was increased to the same degree by stably transfecting the HDAC6 expression vector into MCF-7 cells. In both cases, the cells changed in appearance from their original round shape to an axon-extended shape, like a neuronal cell. This HDAC6 accumulation caused the deacetylation of alpha-tubulin. Either the selective estrogen receptor modulator tamoxifen (TAM) or the pure antiestrogen ICI 182,780 prevented estradiol-induced HDAC6 accumulation and deacetylation of alpha-tubulin, leading to reduced cell motility. Tubacin, an inhibitory molecule that binds to the tubulin deacetylation domain of HDAC6, also prevented estradiol-stimulated cell migration. Finally, we evaluated HDAC6 protein expression in 139 consecutively archived human breast cancer tissues by immunohistochemical staining. The prognostic analyses for these patients revealed no significant differences based on HDAC6 expression. However, subset analysis of ER-positive patients who received adjuvant treatment with TAM (n = 67) showed a statistically significant difference in relapse-free survival and overall survival in favor of the HDAC6-positive group (P < 0.02 and P < 0.05, respectively). HDAC6 expression was an independent prognostic indicator by multivariate analysis (odds ratio = 2.82, P = 0.047). These results indicate the biological significance of HDAC6 regulation via estrogen signaling. PMID- 15806143 TI - High levels of cytoplasmic HTLV-1 Tax mutant proteins retain a Tax-NF-kappaB-CBP ternary complex in the cytoplasm. AB - The oncogenic potential of HTLV-1 Tax protein is partially ascribed to its capacity to activate NF-kappaB. The current view is that Tax acts first in the cytoplasm to dissociate NF-kappaB factors from the IkappaB proteins and enable their nuclear translocation, then Tax links p65(RelA), within the nucleus, to CBP/p300 and P/CAF, which are essential for its optimal transcriptional activity. Our present study challenges the paradigm that Tax-p65(RelA)-CBP/p300 assembly occurs in the nucleus. Using Tax mutants defective for nuclear localization we show that at low levels these mutants induce the nuclear translocation of NF kappaB factors but not their transcriptional activity, whereas at high levels they trap CBP and free p65(RelA) in the cytoplasm and block, thereby, their transcriptional function. In contrast, wild-type (w.t.) Tax strongly stimulated NF-kappaB-dependent gene expression in all tested experimental settings. These data suggest that the Tax-p65(RelA)-CBP ternary complex is established in the cytoplasm rather than in the nucleus. When this complex is formed with w.t. Tax, the entire moiety translocates into the nucleus and exerts high transcriptional activity. However, if the complex is formed with the cytoplasmic Tax mutants, the resulting moiety is retained in the cytoplasm and is, therefore, devoid of transcriptional activity. PMID- 15806144 TI - Characterization of murine S-endoglin isoform and its effects on tumor development. AB - Endoglin is a transmembrane glycoprotein that acts as an auxiliary receptor for transforming growth factor-beta (TGF-beta) and modulates cellular responses to this pleiotropic cytokine. Endoglin is strongly expressed in endothelial cells, where it appears to exert a crucial role in vascular development and angiogenesis. Two endoglin isoforms (L and S), differing in their cytoplasmic domains, have been previously characterized in human tissues. We now demonstrate the existence of similar L- and S-endoglin variants in murine tissues with 47 and 35 amino acids, respectively, in their cytoplasmic tail. RT-PCR analysis showed that L is the predominant endoglin isoform expressed in mouse tissues, although S endoglin mRNA is significantly expressed in liver and lung, as well as in endothelial cell lines. Furthermore, a protein of size equivalent to recombinant S-endoglin expressed in mammalian cells was detected in mouse endothelial cells by Western blot analysis. L- and S-endoglin isoforms can form disulfide-linked heterodimers, as demonstrated by cotransfection of L- and S-endoglin constructs. To address the role of S-endoglin in vivo, an S-Eng(+) transgenic mouse model that targets S-endoglin expression to the endothelium was generated. The lethal phenotype of endoglin-null (Eng(-/-)) mice was not rescued by breeding S-Eng(+) transgenic mice into the endoglin-null background. S-Eng(+) mice exhibited reduced tumor growth and neovascularization after transplantation of Lewis lung carcinoma cells. In addition, S-Eng(+) mice showed a drastic inhibition of benign papilloma formation when subjected to two-stage chemical skin carcinogenesis. These results point to S-endoglin as an antiangiogenic molecule, in contrast to L endoglin which is proangiogenic. Oncogene (2005) 24, 4450-4461. doi:10.1038/sj.onc.1208644 Published online 4 April 2005. PMID- 15806145 TI - Differences in the association of p53 phosphorylated on serine 15 and key enzymes of homologous recombination. AB - Phosphorylation of p53 on serine 15 by ATM or ATR is a frequent modification and initiates a cascade of post-translational modifications. To identify possible mechanisms that modulate p53 functions in recombination surveillance, we compared the nuclear localization of p53 phosphorylated on serine 15 (p53pSer15) and the key enzymes of homologous recombination (HR) after replication fork stalling. We demonstrate an almost mutually exclusive subcompartmentalization with Rad52, while p53pSer15 was colocalizing with 40-60% of the Rad51 and Mre11 foci. Therefore, possible sites of p53pSer15-dependent regulation seem to be sites of Rad51- rather than Rad52-dependent HR processes. Remarkably, the association of p53pSer15 with repair complexes containing Rad51 or Mre11 was transient, because less than 20% of the Rad51 and Mre11 foci overlapped with p53pSer15 after 6 h. When we examined colocalization and co-immunoprecipitation of p53pSer15 and the RecQ helicase BLM with recombination surveillance and proapoptotic functions, we observed colocalization within a fraction of approximately 70% of the BLM foci and stable physical interactions until 6 h after replication arrest. Our data suggest that p53pSer15 plays a dual role in the functional interactions with early complexes of Rad51-dependent recombination and with BLM-associated surveillance and signalling complexes within distinct nuclear subcompartments. PMID- 15806146 TI - Expression of DLK1 in hematopoietic cells results in inhibition of differentiation and proliferation. AB - The Delta-like (DLK1) gene is overexpressed in CD34+ cells from myelodysplastic syndrome (MDS) patients. DLK1 encodes an EGF-like homeotic transmembrane protein homologous to the notch/delta/serrate family. Although exogenous DLK1 promotes maintenance of murine hematopoietic stem cells, the functional effects of DLK1 overexpression in hematopoietic cells are unknown. We show that ectopically expressed DLK1 significantly inhibits differentiation and proliferation of human promyelocytic HL-60 cells. Unlike preadipocytes, where proteolytic processing of membrane-bound protein and release of a soluble form mediates differentiation inhibition, proteolytic release of the extracellular domain was not required for inhibition of hematopoietic cell differentiation. However, intracellular domain interactions were critical to this DLK1 function. We conclude that DLK1 overexpression in hematopoietic cells has important functional consequences. Our studies identify novel molecular mechanisms and indicate that DLK1 has activity both as a soluble and a transmembrane expressed protein. Our results support further investigation of the role of DLK1 in abnormal hematopoiesis in MDS. PMID- 15806147 TI - The second loop of occludin is required for suppression of Raf1-induced tumor growth. AB - Tight junctions (TJs) regulate epithelial cell polarity and paracellular permeability. Loss of functional TJs is commonly associated with epithelial cell derived cancers. Raf1-mediated transformation of rat salivary gland epithelial cells (Pa4-Raf1) induces transcriptional downregulation of the TJ protein occludin and forced re-expression of occludin rescues polarized phenotype of epithelial cells. In the present study, we used this model to examine how specific structural modifications in the occludin protein affect its function in vitro and influence tumor growth in vivo. Our results revealed that neither the C terminal nor the N-terminal half of occludin alone were sufficient to rescue cells from transformation by Raf1. However, forced expression of an occludin mutant lacking the first extracellular loop was sufficient to rescue cells from Raf1-mediated transformation. Interestingly, forced expression of an occludin mutant lacking the second extracellular loop did not rescue the epithelial phenotype in vitro nor did it prevent tumor growth in vivo. These results demonstrate that the TJ protein occludin has a potent inhibitory effect on the Raf1-mediated tumorigenesis, and the second extracellular loop of occludin appears to be critical for this function. PMID- 15806148 TI - Junctional recruitment of mammalian Scribble relies on E-cadherin engagement. AB - Members of the LAP protein family, LET-413 in Caenorhabditis elegans, Scribble in Drosophila melanogaster, and Erbin, Lano, Densin-180 and hScrib in mammals, have conserved structural features. LET-413 and Scribble are junctional proteins involved in establishing and maintaining epithelial cell polarity. scribble also behaves as a neoplastic tumor suppressor gene. We show here that, in epithelial cells, hScrib is recruited at cell-cell junctions in an E-cadherin-dependent manner as shown by calcium switch assays in MDCK cells, re-expression of E cadherin in MDA-231 cells treated by 5-Aza-2'-deoxycytidine (5Aza), and siRNA experiments. hScrib is restricted at the basolateral membrane of epithelial cells by its LRR domain, and is enriched in Triton X-100-insoluble fractions. In breast cancers, most lobular tumors did not express hScrib and E-cadherin while ductal tumors had a less frequent downregulation of hScrib. Our data provide additional insights on the modalities of recruitment of hScrib at the cell-cell junctions, and establish a potential link between the E-cadherin and hScrib tumor suppressors. PMID- 15806149 TI - The Ski oncoprotein is upregulated and localized at the centrosomes and mitotic spindle during mitosis. AB - Ski is an oncoprotein that represses transforming growth factor-beta and nuclear receptor signaling. Despite evidence that relates increased Ski protein levels directly with tumor progression in human cells, the signaling pathways that regulate Ski expression are mostly unidentified. Here we show that the Ski protein levels vary throughout the cell cycle, being lowest at G0/G1. This reduction in Ski protein levels results from proteosomal degradation as suggested by in vivo ubiquitination of Ski and the effects of proteosomal inhibitors. In contrast, an upregulation of the Ski protein was observed in cells going through mitosis. At this stage, we also found that Ski is phosphorylated. In vitro and in vivo data suggest that the phosphorylation of Ski in mitosis is carried out by the main kinase controlling the progression of mitosis, namely cdc2/cyclinB. Interestingly, immunofluorescence experiments, supported by biochemical data, show not only an increase in the Ski protein levels, but also a dramatic redistribution of Ski to the centrosomes and mitotic spindle throughout mitosis. Studies to date on Ski have focused on its role as a transcriptional regulator. However, Ski's increased level and specific relocalization during mitosis suggest that Ski might play a distinct role during this particular phase of the cell cycle. PMID- 15806150 TI - Hepatitis B virus-related insertional mutagenesis in chronic hepatitis B patients as an early drastic genetic change leading to hepatocarcinogenesis. AB - Growing evidence demonstrates that hepatitis B virus (HBV) integration and resulting insertional mutagenesis play an important role in cell growth or maintenance in hepatocellular carcinomas (HCCs). To determine if HBV integration occurs and affects cellular genes at such a stage of infection, we analysed viral host junctions in chronic hepatitis tissues without HCC using PCR amplification with primers specific to human Alu-repeat and HBV. We obtained 42 independent viral-host junctions from six patients examined and identified chromosomal locations for 20 of the 42 junctions. In six clones, each integration apparently affected a single gene. These six candidate genes included one known tumor suppressor gene, three human homologs of drosophila genes that are critical for organ development, one putative oncogene and one recently found chemokine. Our data, together with previously reported HBV integrants in HCCs, suggested preferential HBV integration into chromosome 3 (P = 0.022). Our virus-tagging approach provided (a) firm evidence of HBV integration in hepatocytes at an early stage of chronic infection and (b) revealed cellular genes possibly affected by HBV integration and potentially involved in early steps of the process leading to carcinogenesis. PMID- 15806151 TI - ERK phosphorylation is linked to VEGFR2 expression and Ets-2 phosphorylation in breast cancer and is associated with tamoxifen treatment resistance and small tumours with good prognosis. AB - Extracellular signal-regulated kinase (ERK)1/2 signalling mediates communication between growth factor receptors and the cell nucleus and has been linked to several key events in the transformation process such as proliferation and invasion. We therefore sought to delineate the degree of phosphorylated ERK1/2 in breast cancer and potential links to upstream receptors such as VEGFR2, ErbB2, downstream targets, such as Ets-2, as well as clinico-pathological parameters, clinical outcome and response to tamoxifen. ERK1/2 phosphorylation was assessed by immunohistochemistry using a phospho-specific ERK1/2 antibody in three breast cancer cohorts including a total of 886 tumours arranged in tissue arrays. Cohort I consisted of 114 patients, cohort II of 248 postmenopausal patients randomized to either 2 years of tamoxifen or no adjuvant treatment and cohort III of 524 patients. Surprisingly, ERK1/2 phosphorylation correlated inversely with tumour size. Phosphorylated ERK1/2 was further associated with the presence of VEGFR2 (cohorts II and III) and the degree of phosphorylated Ets-2, indicating in vivo, a signalling cascade from VEGFR2 via ERK1/2 to Ets-2 phosphorylation. Interestingly, ERK1/2 phosphorylation correlated with better survival in untreated patients independently of lymph-node status and tumour size indicating that ERK1/2 signalling might be associated with a less aggressive phenotype. Finally, patients with oestrogen receptor positive and ERK1/2 phosphorylated tumours also had an impaired tamoxifen response. PMID- 15806152 TI - The antiepidermal growth factor receptor monoclonal antibody cetuximab/C225 reduces hypoxia-inducible factor-1 alpha, leading to transcriptional inhibition of vascular endothelial growth factor expression. AB - We have previously shown that the antiepidermal growth factor receptor monoclonal antibody cetuximab (C225; Erbitux), which was recently approved for the treatment of metastatic colorectal cancer, has antiangiogenic properties, inhibiting vascular endothelial growth factor (VEGF) secretion in culture and in animal models. Here, we have furthered the study by demonstrating that cetuximab reduces cellular levels of hypoxia-inducible factor-1 alpha (HIF-1alpha), a transcriptional regulator of VEGF expression, in A431 epidermoid carcinoma cells under both normoxic and hypoxic culture conditions. Expression of a constitutively active Ras in A431 cells rendered cellular resistance to the cetuximab-mediated reduction of the HIF-1alpha level. Cell lines with naturally occurring phosphatase and tensin homologue deleted on chromosome 10 mutations or deletions were also resistant to cetuximab-mediated reduction of the HIF-1alpha level. Pharmacologic inhibition of phosphatidylinositol 3-kinase with LY294002 reduced the HIF-1alpha level in both normoxic and hypoxic A431 cells, whereas inhibition of the mitogen-activated protein kinase kinase by PD98059 reduced the level of HIF-1alpha only in normoxic A431 cells. In addition, cetuximab reduced the cellular level of HIF-1alpha in the presence of a proteasome inhibitor, lactacystin, indicating that cetuximab acts mainly at the level of protein synthesis. The reduction of HIF-1alpha in response to cetuximab treatment was accompanied by transcriptional inhibition of VEGF expression, measured by a luciferase assay in A431 cells transfected with a vector containing the VEGF hypoxia response element. Taken together, our results indicate that the previously demonstrated inhibition of VEGF by cetuximab occurs at the level of transcription in response to a reduced level of HIF-1alpha and justify further testing of therapeutic strategies that combine cetuximab with approaches inhibiting the function of VEGF or the VEGF receptor. PMID- 15806153 TI - Estrogen-induced loss of progesterone receptor expression in normal and malignant ovarian surface epithelial cells. AB - While estrogens are suspected risk factors for epithelial ovarian cancer (OCa), progesterone (P4) has been shown to exert protective effects. The biological actions of P4 in target cells are mediated by progesterone receptors (PRs) that exist principally as A- and B-isoforms. We observed overexpression of PR-A and PR B protein in two lines of OCa cells when compared to two lines of nontumorigenic, normal human ovarian surface epithelial (HOSE) cells. Treatment of HOSE or OCa cells with estrone or 17beta-estradiol at 10(-8) M for a period of 72 h induced significant loss of PR-A and PR-B mRNA and protein expression, with the regulation primarily controlled at the transcriptional level. In contrast, breast cancer cells (line MCF-7) exposed to estrogens upregulated PR-A and PR-B expression. Of significance, both the inhibitory and stimulatory actions of estrogens were blocked by the specific ER-antagonist ICI 182,780 (ICI, 10(-5) M), confirming estrogen specificity. Co-treatment of estrogen-exposed HOSE, OCa, and MCF-7 cell lines with inhibitors of type 1- and type 2-17beta hydroxysteroid dehydrogenase did not affect the previously observed changes in PR expression, suggesting that the action of each estrogen is direct and not mediated via conversion to its metabolic counterpart. Green fluorescence protein (GFP)-PR-A and GFP-PR-B were localized in the cytoplasmic compartment of untreated HOSE cells and translocated to the nucleus after P4 treatment, while both chimera PRs resided in the nuclei of OCa cells in a ligand-independent manner. In OCa cell cultures, P4 (10(-6) M), but not RU486 (10(-5) M), induced apoptosis that was blocked by co-treatment with the antiprogestin but enhanced by co-treatment with ICI. In sharp contrast, P4 induced proliferation, while ICI and RU486 caused cell death in MCF-7 cells. In conclusion, this study is first to demonstrate estrogens as negative regulators of PR expression in HOSE/OCa cells and to provide a mechanistic basis upon which to explain the antagonism of estrogens on the anti OCa action of progestins. It also raises the possibility of using progestin and ICI as a combinational therapy for OCa treatment. PMID- 15806154 TI - The glucose dependence of Akt-transformed cells can be reversed by pharmacologic activation of fatty acid beta-oxidation. AB - Activation of the oncogenic kinase Akt stimulates glucose uptake and metabolism in cancer cells and renders these cells susceptible to death in response to glucose withdrawal. Here we show that 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) reverses the sensitivity of Akt-expressing glioblastoma cells to glucose deprivation. AICAR's protection depends on the activation of AMPK, as expression of a dominant-negative form of AMPK abolished this effect. AMPK is a cellular energy sensor whose activation can both block anabolic pathways such as protein synthesis and activate catabolic reactions such as fatty acid oxidation to maintain cellular bioenergetics. While rapamycin treatment mimicked the effect of AICAR on inhibiting markers of cap-dependent translation, it failed to protect Akt-expressing cells from death upon glucose withdrawal. Compared to control cells, Akt-expressing cells were impaired in the ability to induce fatty acid oxidation in response to glucose deprivation unless stimulated with AICAR. Stimulation of fatty acid oxidation was sufficient to maintain cell survival as activation of fatty acid oxidation with bezafibrate also protected Akt-expressing cells from glucose withdrawal-induced death. Conversely, treatment with a CPT-1 inhibitor to block fatty acid import into mitochondria prevented AICAR from stimulating fatty acid oxidation and promoting cell survival in the absence of glucose. Finally, cell survival did not require reversal of Akt's effects on either protein translation or lipid synthesis as the addition of the cell penetrant oxidizable substrate methyl-pyruvate was sufficient to maintain survival of Akt-expressing cells deprived of glucose. Together, these data suggest that activation of Akt blocks the ability of cancer cells to metabolize nonglycolytic bioenergetic substrates, leading to glucose addiction. PMID- 15806155 TI - CXCR4 chemokine receptor and integrin signaling co-operate in mediating adhesion and chemoresistance in small cell lung cancer (SCLC) cells. AB - Small cell lung cancer (SCLC) is an aggressive, rapidly metastazising neoplasm with a high propensity for marrow involvement. SCLC cells express high levels of functional CXCR4 receptors for the chemokine stromal-cell-derived factor-1 (SDF 1/CXCL12). Adhesion of SCLC cells to extracellular matrix or accessory cells within the tumor microenvironment confers resistance to chemotherapy via integrin signaling and thus may be responsible for residual disease and relapses commonly seen in SCLC. We examined the signaling mechanisms that regulate CXCL12-induced adhesion of SCLC cells to fibronectin, collagen, and stromal cells and the effects on SCLC cell chemoresistance. We found that CXCL12-induced integrin activation which resulted in an increased adhesion of SCLC cells to fibronectin and collagen. This was mediated by alpha2, alpha4, alpha5, and beta1 integrins along with CXCR4 activation, which could be inhibited by CXCR4 antagonists. Stromal cells protected SCLC cells from chemotherapy-induced apoptosis, and this protection could also be antagonized by CXCR4 inhibitors. We conclude that activation of integrins and CXCR4 chemokine receptors co-operate in mediating adhesion and survival signals from the tumor microenvironment to SCLC cells. Therefore, CXCR4 antagonists in combination with cytotoxic drugs should be explored in SCLC to overcome CXCL12-mediated adhesion and survival signals in the tumor microenvironment. PMID- 15806156 TI - p53 deficiency does not affect mutation rate in the mouse germline. AB - Although the influence of p53 deficiency on somatic genetic stability is well established, its effect in the germline is poorly understood. Mutation rates at two expanded simple tandem repeat (ESTR) loci were studied in the germline of nonexposed and irradiated p53-deficient mice. Spontaneous mutation rates in the homozygous and heterozygous p53-deficient males did not significantly differ from that in the isogenic wild-type mice. Acute exposure to 1 Gy of X rays resulted in a similar increase in mutation rates across males with different p53 genotypes. ESTR mutation spectra did not significantly differ across males with different p53 genotypes. Taken together, these results suggest that p53-deficiency does not affect spontaneous and radiation-induced mutation in the mouse germline. PMID- 15806157 TI - A functional heparan sulfate mimetic implicates both heparanase and heparan sulfate in tumor angiogenesis and invasion in a mouse model of multistage cancer. AB - Heparan sulfate proteoglycans are integral components of the extracellular matrix that surrounds all mammalian cells. In addition to providing structural integrity, they act as a storage depot for a variety of heparan sulfate (HS) binding proteins, including growth factors and chemokines. Heparanase is a matrix degrading enzyme that cleaves heparan sulfate side chains from the core proteoglycans, thus liberating such HS-binding proteins, as well as potentially contributing to extracellular matrix degradation. Here, we report that heparanase mRNA and protein expression are increased in the neoplastic stages progressively unfolding in a mouse model of multistage pancreatic islet carcinogenesis. Notably, heparanase is delivered to the neoplastic lesions in large part by infiltrating Gr1+/Mac1+ innate immune cells. A sulfated oligosaccharide mimetic of heparan sulfate, PI-88, was used to inhibit simultaneously both heparanase activity and HS effector functions. PI-88 had significant effects at distinct stages of tumorigenesis, producing a reduction in the number of early progenitor lesions and an impairment of tumor growth at later stages. These responses were associated with decreased cell proliferation, increased apoptosis, impaired angiogenesis, and a substantive reduction in the number of invasive carcinomas. In addition, we show that the reduction in tumor angiogenesis is correlated with a reduced association of VEGF-A with its receptor VEGF-R2 on the tumor endothelium, implicating heparanase in the mobilization of matrix-associated VEGF. These data encourage clinical applications of inhibitors such as PI-88 for the many human cancers where heparanase expression is elevated or mobilization of HS-binding regulatory factors is implicated. PMID- 15806158 TI - Distinct molecular phenotype of malignant CD34(+) hematopoietic stem and progenitor cells in chronic myelogenous leukemia. AB - Chronic myelogenous leukemia (CML) is a malignant disorder of the hematopoietic stem cell characterized by the BCR-ABL oncogene. We examined gene expression profiles of highly enriched CD34(+) hematopoietic stem and progenitor cells from patients with CML in chronic phase using cDNA arrays covering 1.185 genes. Comparing CML CD34(+) cells with normal CD34(+) cells, we found 158 genes which were significantly differentially expressed. Gene expression patterns reflected BCR-ABL-induced functional alterations such as increased cell-cycle and proteasome activity. Detoxification enzymes and DNA repair proteins were downregulated in CML CD34(+) cells, which might contribute to genetic instability. Decreased expression of junction plakoglobulin and CXC chemokine receptor 4 (CXCR-4) might facilitate the release of immature precursors from bone marrow in CML. GATA-2 was upregulated in CML CD34(+) cells, suggesting an increased self-renewal in comparison with normal CD34(+) cells. Moreover, we found upregulation of the proto-oncogene SKI and of receptors for neuromediators such as opioid mu1 receptor, GABA B receptor, adenosine A1 receptor, orexin 1 and 2 receptors and corticotropine-releasing hormone receptor. Treatment of CML progenitor cells with the selective adenosine A1 receptor antagonist 8 cyclopentyl-1,3-dipropylxanthine (DPCPX) resulted in a dose-dependent significant inhibition of clonogenic growth by 40% at a concentration of 10(-5) M, which could be reversed by the equimolar addition of the receptor agonist 2-chloro-N6 cyclopentyladenosine (P<0.05). The incubation of normal progenitor cells with DPCPX resulted in an inhibition of clonogenic growth to a significantly lesser extent in comparison with CML cells (P<0.05), suggesting that the adenosine A1 receptor is of functional relevance in CML hematopoietic progenitor cells. PMID- 15806159 TI - The adaptor Grb7 is a novel calmodulin-binding protein: functional implications of the interaction of calmodulin with Grb7. AB - We demonstrate using Ca2+-dependent calmodulin (CaM)-affinity chromatography and overlay with biotinylated CaM that the adaptor proteins growth factor receptor bound (Grb)7 and Grb7V (a naturally occurring variant lacking the Src homology 2 (SH2) domain) are CaM-binding proteins. Deletion of an amphiphilic basic amino acid sequence (residues 243-256) predicted to form an alpha-helix located in the proximal region of its pleckstrin homology (PH) domain demonstrates the location of the CaM-binding domain. This site is identical in human and rodents Grb7, and shares great homology with similar regions of Grb10 and Grb14, and the Mig10 protein from Caenorhabditis elegans. We show that Grb7 and Grb7V are present in the cytosol and bound to membranes, while the deletion mutants (Grb7Delta and Grb7VDelta) have less capacity to be associated to membranes. Grb7Delta maintains in part the capacity to bind phosphoinositides, and CaM competes for phosphoinositide binding. Activation of ErbB2 by heregulin beta1 decreases the pool of Grb7 associated to membranes. The cell-permeable CaM antagonist W7 (N-(6 aminohexyl)-5-chloro-1-naphthalenesulfonamide), but not the CaM-dependent protein kinase II inhibitor KN93, prevents this effect. Highly specific cell-permeable CaM inhibitory peptides decrease the association of Grb7 to membranes. This suggests that CaM regulates the intracellular mobilization of Grb7 in living cells. Direct interaction between enhanced yellow fluorescent protein (EYFP)-Grb7 and enhanced cyan fluorescent protein (ECFP)-CaM chimeras at the plasma membrane of living cells was demonstrated by fluorescence resonance energy transfer (FRET). The FRET signal dramatically decreased in cells loaded with a cell permeable Ca2+ chelator, and was significantly attenuated when enhanced yellow fluorescent protein-Grb7 chimera (EYFP-Grb7)Delta instead of EYFP-Grb7 was used. Finally, we show that conditioned media from cells transiently transfected with Grb7Delta and Grb7VDelta lost its angiogenic activity, in contrast to those from cells transiently transfected with their wild-type counterparts. PMID- 15806160 TI - Akt phosphorylates the Y-box binding protein 1 at Ser102 located in the cold shock domain and affects the anchorage-independent growth of breast cancer cells. AB - Akt/PKB is a serine/threonine kinase that promotes tumor cell growth by phosphorylating transcription factors and cell cycle proteins. There is particular interest in finding tumor-specific substrates for Akt to understand how this protein functions in cancer and to provide new avenues for therapeutic targeting. Our laboratory sought to identify novel Akt substrates that are expressed in breast cancer. In this study, we determined that activated Akt is positively correlated with the protein expression of the transcription/translation factor Y-box binding protein-1 (YB-1) in primary breast cancer by screening tumor tissue microarrays. We therefore questioned whether Akt and YB-1 might be functionally linked. Herein, we illustrate that activated Akt binds to and phosphorylates the YB-1 cold shock domain at Ser102. We then addressed the functional significance of disrupting Ser102 by mutating it to Ala102. Following the stable expression of Flag:YB-1 and Flag:YB-1 (Ala102) in MCF-7 cells, we observed that disruption of the Akt phosphorylation site on YB-1 suppressed tumor cell growth in soft agar and in monolayer. This correlated with an inhibition of nuclear translocation by the YB-1(Ala102) mutant. In conclusion, YB-1 is a new Akt substrate and disruption of this specific site inhibits tumor cell growth. PMID- 15806161 TI - Somatic heterozygous mutations in ETV6 (TEL) and frequent absence of ETV6 protein in acute myeloid leukemia. AB - ETV6 (ets translocation variant gene 6) TEL (translocation ets leukemia), encoding a transcriptional repressor, is involved in various translocations associated with human malignancies. Strikingly, the nonrearranged ETV6 allele is often deleted or inactivated in cells harboring these translocations. Although ETV6 translocations are infrequent in acute myeloid leukemia (AML), mutations or deregulated expression of ETV6 may contribute to leukemogenesis. To investigate the involvement of ETV6 in AML, we analysed 300 newly diagnosed patients for mutations in the coding region of the gene. Furthermore, we studied protein expression in 77 patients using two ETV6-specific antibodies. Five somatic heterozygous mutations were detected, which affected either the homodimerization- or the DNA-binding domain of ETV6. The proteins translated from the cDNAs of these mutants were unable to repress transcription and showed dominant-negative effects. In addition, we demonstrate that one-third of AML patients have deficient ETV6 protein expression, which is not related to ETV6 mRNA expression levels. In conclusion, we demonstrate that ETV6 abnormalities are not restricted to translocations and occur more frequently in AML than previously thought. Additional comprehensive studies are required to define the clinical consequence of ETV6 loss of function in AML. PMID- 15806162 TI - Mitogen regulated induction of FRA-1 proto-oncogene is controlled by the transcription factors binding to both serum and TPA response elements. AB - FRA-1, a member of the FOS family of transcription factors, is overexpressed in a variety of human tumors, and contributes to tumor progression. In addition to mitogens, various toxicants and carcinogens persistently induce FRA-1 expression in vitro and in vivo. Although the mitogen induced expression of c-FOS is relatively well understood, it is poorly defined in the case of FRA-1. Our recent analysis of the FRA-1 promoter has shown a critical role for a TRE located at 318 in mediating the TPA-induced expression. The -379 to -283 bp promoter segment containing a critical TRE (-318), however, is insufficient for the induction of FRA-1 promoter. Here, we show that a 40-bp (-276/-237) segment, comprising a TCF binding site and the CArG box (collectively known as serum response element, SRE), and an ATF site, is also necessary for the FRA-1 induction by TPA and EGF. Interestingly, the -283 to +32 bp FRA-1 promoter fragment containing an SRE and an ATF site alone was also insufficient to confer TPA sensitivity to a reporter gene. However, in association with the -318 TRE, the SRE and ATF sites imparted a strong TPA-inducibility to the reporter. Similarly, EGF also required these motifs for the full induction of this gene. Using ChIP assays we show that, in contrast to c-Jun, SRF, Elk1, ATF1 and CREB proteins bind to SRE and ATF sites of the FRA-1 promoter, constitutively. RNAi-mediated knockdown of endogenous SRF, ELK1 and c-JUN protein expression significantly reduced TPA-stimulated FRA-1 promoter activity. Thus, a bipartite enhancer formed by an upstream TRE and the downstream SRE and ATF sites and the cognate factors is necessary and sufficient for the regulation of FRA-1 in response to mitogens. PMID- 15806163 TI - TRAIL sensitizes for ionizing irradiation-induced apoptosis through an entirely Bax-dependent mitochondrial cell death pathway. AB - The death ligand TRAIL has been suggested as a suitable biological agent for the selective induction of cell death in cancer cells. Moreover, TRAIL synergizes with DNA-damaging therapies such as chemotherapeutic drugs or ionizing irradiation (IR). Here, we show that synergy of TRAIL and IR, that is, crosssensitization between TRAIL and IR for induction of apoptosis, entirely depends on Bax proficiency in human DU145 and HCT116 carcinoma cells. DU145 prostate carcinoma cells that have lost Bax protein expression due to mutation fail to activate caspase-3 and -9 when exposed to TRAIL and IR. In contrast, TRAIL sensitized for IR-induced apoptosis and vice versa upon reconstitution of Bax expression. Notably, both DU145 and HCT116 still express significant levels of the multidomain proapoptotic Bcl-2 homolog Bak. This indicates that Bak is not sufficient to mediate crosssensitization and synergism between IR and TRAIL. These data clearly establish distinct roles for Bax and Bak in linking the TRAIL death receptor pathway to the mitochondrial apoptosis signaling cascade upon DNA damage by IR. PMID- 15806164 TI - Gene profiling reveals specific oncogenic mechanisms and signaling pathways in oncocytic and papillary thyroid carcinoma. AB - The oncogenic pathways in mitochondrial-rich thyroid carcinomas are not clearly understood. To investigate the possible implication of mitochondrial abundance in the genesis of thyroid tumors, we have explored the gene expression profile of six oncocytic carcinomas and six mitochondrial-rich papillary carcinomas using cDNA-microarray technology. A supervised approach allowed us to identify 83 genes differentially expressed in the two types of carcinoma. These genes were classified according to their ontologic profiles. Three genes, NOS3, alpha actinin-2 and alpha-catenin, suspected of playing a role in tumor genesis, were explored by quantitative RT-PCR analysis and immunohistochemistry. Of the 59 genes overexpressed in papillary carcinomas, 51% were involved in cell communication. Of the 24 genes overexpressed in oncocytic carcinomas, 84% were involved in mitochondrial and cellular metabolism. Our results suggest that mitochondrial respiratory chain complexes III and IV play a significant role in the regulation of reactive oxygen species production by oncocytic tumors. PMID- 15806165 TI - Role of the adenosine 3',5'-cyclic monophosphate (cAMP) in enhancing the efficacy of siRNA-mediated gene silencing in neuroblastoma cells. AB - Gene-silencing activity mediated by siRNA has been demonstrated in mammalian cells; however, the mechanism of its regulation is not well understood. Since downregulation of a number of genes occurs during adenosine 3',5'-cyclic monophosphate (cAMP)-induced differentiation of neuroblastoma (NB) cells, it is possible that cAMP may play a role in regulating siRNA activity during differentiation. To study this, we utilized an NB cell line (NBP2-PN25) that expresses a short-lived green fluorescent protein (d2EGFP) under the CMV promoter. These cells were transfected with a retroviral plasmid that expresses U6 promoter-driven expression of siRNA targeted to d2EGFP and then were treated with cAMP-elevating agents (200 microg/ml RO20-1724, an inhibitor of cyclic nucleotide phosphodiesterase, and 1 microg/ml prostaglandin A1, a stimulator of adenylate cyclase) for 2 or 24 h. The siRNA activity was measured by determining the level of intensity of d2EGFP protein by flow cytometry, and the level of d2EGFP mRNA by real-time PCR. The results showed that cAMP-elevating agents enhanced U6-driven siRNA activity directed towards d2EGFP in NB cells 24 h after treatment. One of the mechanisms of action of cAMP is mediated via phosphatidylinositol 3-kinase (PI3K) inhibition; therefore, we have investigated the effect of a PI3K inhibitor on siRNA activity. This study showed that inhibition of PI3K also enhanced U6-driven siRNA activity towards d2EGFP. cAMP stimulating agents increased U6 transcript levels, perhaps suggesting that increased siRNA activity may in part be due to an increase in transcriptional activity. When NB cells were transfected with a synthetic siRNA directed to d2EGFP, both cAMP elevation and PI3K inhibition similarly enhanced siRNA activity. Sodium butyrate, which inhibits the growth of NB cells similar to the effect produced by cAMP, did not affect U6-driven siRNA activity towards d2EGFP. Protein kinase C (PKC) activation or inhibition also failed to affect siRNA activity in NB cells. This study also showed that cAMP elevation and PI3K inhibition increases U6-driven siRNA activity directed towards an endogenous gene, p53. Our data suggest a role for the cAMP pathway in affecting the efficacy of siRNA system during differentiation of NB cells. PMID- 15806166 TI - Kruppel-like factor 4 prevents centrosome amplification following gamma irradiation-induced DNA damage. AB - Centrosome duplication is a carefully controlled process in the cell cycle. Previous studies indicate that the tumor suppressor, p53, regulates centrosome duplication. Here, we present evidence for the involvement of the mammalian Kruppel-like transcription factor, KLF4, in preventing centrosome amplification following DNA damage caused by gamma-irradiation. The colon cancer cell line HCT116, which contains wild-type p53 alleles (HCT116 p53+/+), displayed stable centrosome numbers following gamma-irradiation. In contrast, HCT116 cells null for the p53 alleles (HCT116 p53-/-) exhibited centrosome amplification after irradiation. In the latter cell line, KLF4 was not activated following gamma irradiation due to the absence of p53. However, centrosome amplification could be suppressed in irradiated HCT116 p53-/- cells by conditional induction of exogenous KLF4. Conversely, in a HCT116 p53+/+ cell line stably transfected with small hairpin RNA (shRNA) designed to specifically inhibit KLF4, gamma irradiation induced centrosome amplification. In these cells, the inability of KLF4 to become activated in response to DNA damage was directly associated with an increase in cyclin E level and Cdk2 activity, both essential for regulating centrosome duplication. Cotransfection experiments showed that KLF4 overexpression suppressed the promoter activity of the cyclin E gene. The results of this study demonstrated that KLF4 is both necessary and sufficient in preventing centrosome amplification following gamma-radiation-induced DNA damage and does so by transcriptionally suppressing cyclin E expression. PMID- 15806167 TI - Cleavage of cten by caspase-3 during apoptosis. AB - Normal endothelial and epithelial cells undergo apoptosis when cell adhesion and spreading are disrupted, implying a critical role of focal adhesions in cell survival. Cten is a focal adhesion molecule of the tensin family. In contrast to other tensins, cten expression is limited to very few tissues, such as the prostate, and only in epithelial cells. Here, we have explored the potential roles of cten in apoptosis. We found cten was cleaved during apoptosis induced by staurosporine in normal prostate epithelial cells. By using recombinant caspases and site-directed mutagenesis, we have identified caspase-3 as the major protease to digest cten at the DSTD(570 downward arrow)S site. The biological relevance of cten-cleaved fragments was demonstrated by cells ectopically expressing these fragments. Cten fragment (residues 571-715) containing the phosphotyrosine binding domain significantly reduced the growth rate. Detection of cleaved poly(ADP-ribose) polymerase and annexin binding in cells expressing cten (571 715) indicated that a fraction of cells underwent apoptosis. These results demonstrate that cten is a target of caspase-3 and the resultant fragments could further promote apoptosis. PMID- 15806168 TI - Gli1 is important for medulloblastoma formation in Ptc1+/- mice. AB - Germline mutations in the human homolog of the patched1 (PTCH1) are associated with basal cell nevus carcinoma syndrome (BCNS or Gorlin syndrome), which is characterized by developmental anomalies, radiation hypersensitivity and a predisposition to medulloblastomas and skin tumors. Patched1 (Ptc1) functions as a receptor for Sonic hedgehog (Shh) in a wide range of biological processes. Binding of Shh to Ptc1 results in activation of Smoothened (Smo), which in turn stimulates expression of downstream target genes including Ptc1 and Gli1. Gli1 is a member of a family of DNA-binding zinc-finger proteins, including Gli2 and Gli3, that function in transcription control. Here, we report that inactivation of both Gli1 alleles in Ptc1+/- mice significantly reduces spontaneous medulloblastoma formation. Therefore, Gli1 is not only a marker of pathway activation but also plays a functional role in medulloblastoma formation. Interestingly, Gli2 levels were elevated in medulloblastoma cells but not in normal granule neuron precursors during cerebellar development in mice lacking Gli1. In cultured fibroblasts, Gli1 was more potent than Gli2 at inducing cell transformation. These results demonstrate that Gli1 plays a central role in medulloblastoma formation in Ptc1+/- mice and that Gli2 may also contribute to oncogenesis. PMID- 15806169 TI - CpG island promoter hypermethylation of a novel Ras-effector gene RASSF2A is an early event in colon carcinogenesis and correlates inversely with K-ras mutations. AB - We report in silico identification and characterisation of a novel member of the ras association domain family 1 (RASSF1)/NORE1 family, namely, RASSF2, located at chromosomal region 20p13. It has three isoforms, all contain a ras association domain in the C-terminus. The longest isoform RASSF2A contains a 5' CpG island. RASSF2A was cloned from a brain cDNA library and directly sequenced, confirming the genomic gene structure. In previous reports, we and others have demonstrated that RASSF1A is epigenetically inactivated in a variety of cancers, including sporadic colorectal cancer (CRC). In the present report, we analysed the methylation status of RASSF2A promoter region CpG island in sporadic CRC and compared it to K-ras mutation status. RASSF2A promoter region CpG island was hypermethylated in a majority of colorectal tumour cell lines (89%) and in primary colorectal tumours (70%), while DNA from matched normal mucosa was found to be unmethylated (tumour-specific methylation). RASSF2A expression was reactivated in methylated tumour cell lines after treatment with 5-aza 2 deoxycytidine. RASSF2A methylation is an early event, detectable in 7/8 colon adenomas. Furthermore, 75% of colorectal tumours with RASSF2A methylation had no K-ras mutations (codons, 12 and 13) (P=0.048), Fisher's exact test). Our data demonstrate that RASSF2A is frequently inactivated in CRCs by CpG island promoter hypermethylation, and that epigenetic (RASSF2A) and genetic (K-ras) changes are mutually exclusive and provide alternative pathways for affecting Ras signalling. PMID- 15806170 TI - Inhibition of endogenous reverse transcriptase antagonizes human tumor growth. AB - Undifferentiated cells and embryos express high levels of endogenous non telomerase reverse transcriptase (RT) of retroposon/retroviral origin. We previously found that RT inhibitors modulate cell growth and differentiation in several cell lines. We have now sought to establish whether high levels of RT activity are directly linked to cell transformation. To address this possibility, we have employed two different approaches to inhibit RT activity in melanoma and prostate carcinoma cell lines: pharmacological inhibition by two characterized RT inhibitors, nevirapine and efavirenz, and downregulation of expression of RT encoding LINE-1 elements by RNA interference (RNAi). Both treatments reduced proliferation, induced morphological differentiation and reprogrammed gene expression. These features are reversible upon discontinuation of the anti-RT treatment, suggesting that RT contributes to an epigenetic level of control. Most importantly, inhibition of RT activity in vivo antagonized tumor growth in animal experiments. Moreover, pretreatment with RT inhibitors attenuated the tumorigenic phenotype of prostate carcinoma cells inoculated in nude mice. Based on these data, the endogenous RT can be regarded as an epigenetic regulator of cell differentiation and proliferation and may represent a novel target in cancer therapy. PMID- 15806171 TI - The fibroblast growth factor binding protein is a novel interaction partner of FGF-7, FGF-10 and FGF-22 and regulates FGF activity: implications for epithelial repair. AB - The fibroblast growth factor-binding protein (FGF-BP) binds and activates FGF-1 and FGF-2, thereby contributing to tumor angiogenesis. In this study, we identified novel binding partners of FGF-BP, and we provide evidence for a role of this protein in epithelial repair processes. We show that expression of FGF-BP increases after injury to murine and human skin, in particular in keratinocytes. This upregulation is most likely achieved by major keratinocyte mitogens present at the wound site. Most importantly, we demonstrate that FGF-BP interacts with FGF-7, FGF-10, and with the recently identified FGF-22, and enhances the activity of low concentrations of ligand. Due to the important functions of FGF-7 and FGF 10 for repair of injured epithelia, our findings suggest that upregulation of FGF BP expression after injury stimulates FGF activity at the wound site, thus enhancing the process of epithelial repair. PMID- 15806172 TI - Viral oncoproteins E1A and E7 and cellular LxCxE proteins repress SUMO modification of the retinoblastoma tumor suppressor. AB - The retinoblastoma tumor suppressor protein (pRB) is a major regulator of cell cycle progression and cellular differentiation. Central to pRB function is the pocket domain, which serves as the main binding region for cellular regulators. In tumors pRB is frequently inactivated by mutations in the pocket domain or by binding of viral oncoproteins to this region. A characteristic feature of these viral oncoproteins and many cellular pRB-binding partners is an LxCxE sequence motif, which interacts with pRB's pocket domain. Here, we show that the ubiquitin like modifier SUMO is covalently attached to a distinct residue (K720) of pRB within the B-box of the pocket region that binds LxCxE-motif proteins. We provide evidence that SUMO preferentially targets the active, hypophosphorylated form of pRB and show that tumorigenic mutations of pRB in the pocket domain lead to a loss of SUMOylation. Notably, the level of pRB SUMOylation is controlled by the interaction of pRB with viral and cellular LxCxE-motif proteins. Inhibitors of pRB function, including the viral oncoproteins E1A and E7 and the cellular E1A like inhibitor of differentiation EID-1, completely abolish SUMO modification of pRB. Conversely, pRB mutants deficient in binding of LxCxE-motif proteins exhibit a drastically enhanced modification by SUMO. Finally, we provide evidence that SUMOylation can influence pRB function, as the SUMO-deficient pRB(K720R) mutant exerts a slightly higher repressive potential on an E2F-responsive reporter gene than wild-type pRB. Taken together, these data identify SUMO modification as a novel post-translational modification of pRB that may control pRB activity by modulating LxCxE-pocket interactions. PMID- 15806173 TI - Positive feedback regulation between AKT activation and fatty acid synthase expression in ovarian carcinoma cells. AB - Activation of AKT and overexpression of fatty acid synthase (FAS) are frequently observed in human ovarian cancer. To explore a possible connection between AKT and FAS, immunohistochemical analyses were conducted on an ovarian cancer tissue microarray, which revealed a significant correlation between phosphorylated AKT (phospho-AKT) and expression of FAS. To investigate the relationship between phospho-AKT and FAS in vitro, a variety of experiments employing a specific phosphatidylinositol 3-OH kinase (PI3K) inhibitor (LY294002), inducible PTEN expression in PTEN-null cells, or AKT1 siRNA demonstrated that phosphatidylinositol-3 kinase (PI3K)/AKT signaling modulates FAS expression. In contrast, inhibition of FAS activity by the drug C75 resulted in downregulation of phospho-AKT and increased cell death. To explore the functional relationship between phospho-AKT and FAS, we used SKOV3, C200, and OVCAR10 ovarian carcinoma cells, which have constitutively active AKT, and OVCAR5 cells, which have very low basal phospho-AKT levels. Treatment with LY294002 abolished AKT activity and potentiated apoptosis induced by FAS inhibitors cerulenin or C75 only in cells with constitutively active AKT, suggesting that constitutive activation of AKT protects against FAS inhibitor-induced cell death. Furthermore, inhibition of FAS activity by cerulenin or C75 resulted in downregulation of phospho-AKT, which preceded the induction of apoptosis. To investigate the relationship between phospho-AKT and FAS in vivo, severe combined immunodeficient mice injected intraperitoneally with SKOV3 cells were treated with C75. Growth of SKOV3 xenografts was markedly inhibited by C75. Analysis of the levels of phospho-AKT and FAS in C75-treated tumors revealed concordant downregulation of phospho-AKT and FAS. Collectively, our findings are consistent with a working model in which AKT activation regulates FAS expression, at least in part, whereas FAS activity modulates AKT activation. PMID- 15806174 TI - HSP25 inhibits radiation-induced apoptosis through reduction of PKCdelta-mediated ROS production. AB - Since radiation-induced caspase-dependent apoptosis and ROS generation were partially prevented by HSP25 overexpression, similar to the treatment of control cells with antioxidant agents such as DPI and tiron, questions arise whether radiation-mediated ROS generation contributes to the apoptotic cell death, and also whether HSP25 overexpression can reduce ROS mediated apoptotic cell death. In the present study, radiation-induced cytochrome c release from mitochondria and activation of caspases accompanied by a decrease of mitochondrial membrane potential in Jurkat T cells were shown to be inhibited by mitochondrial complex I inhibitor rotenone, suggesting that mitochondrial ROS might be important in radiation-induced caspase-dependent apoptosis. When HSP25 was overexpressed, effects similar to the treatment of cells with the antioxidants were obtained, indicating that HSP25 suppressed radiation-induced mitochondrial alteration that resulted in apoptosis. Furthermore, activation of p38 MAP kinase by radiation was associated with radiation-induced cell death and ROS production and PKCdelta was an upstream molecule for p38 MAP kinase activation, ROS generation and subsequent caspase-dependent apoptotic events. However, in the HSP25 overexpressed cells, the above-described effects were blocked. In fact, radiation-induced membrane translocation of PKCdelta and tyrosine phosphorylation were inhibited by HSP25. Based on the above data, we suggest that HSP25 downregulates PKCdelta, which is a key molecule for radiation-induced ROS generation and mitochondrial-mediated caspase-dependent apoptotic events. PMID- 15806175 TI - Increased prevalence of the BRCA2 polymorphic stop codon K3326X among individuals with familial pancreatic cancer. AB - Germline BRCA2 mutations predispose to the development of pancreatic cancer. A polymorphic stop codon in the coding region of BRCA2 (K3326X) has been described, and although an initial epidemiological study suggested it was not disease causing, subsequent studies have been inconclusive. To investigate the biological significance of the K3326X polymorphism, we determined its prevalence in patients with sporadic and familial pancreatic cancer. Using a case-control design, we studied 250 patients with resected sporadic pancreatic adenocarcinomas, 144 patients with familial pancreatic adenocarcinoma, 115 spouses of patients with pancreatic cancer, and a disease control group of 135 patients without a personal history of cancer who had undergone cholecystectomy for non-neoplastic disease. The K3326X polymorphism was detected using heteroduplex analysis and DNA sequencing. The BRCA2 K3326X polymorphism was significantly more prevalent in individuals with familial pancreatic cancer: 8/144 (5.6%) vs 3/250 controls (1.2%) (odds ratio, 4.84; 95% CI, 1.27-18.55, P<0.01). One K3326X carrier with familial pancreatic cancer carried an alteration (IVS 16-2A>G) suspected to be deleterious. Excluding this case did not alter the significance of the association (OR: 4.24, P<0.01). In contrast, there was no difference in prevalence among individuals with sporadic pancreatic cancer - 7/250 (OR: 2.37, 95% CI: 0.61-9.27). The increased prevalence of the BRCA2 K3326X polymorphism in patients with familial pancreatic cancer suggests that this polymorphism is deleterious and contributes to pancreatic cancer risk. PMID- 15806176 TI - Modulation of CDK2-AP1 (p12(DOC-1)) expression in human colorectal cancer. AB - We have previously demonstrated an association between microsatellite instability and decreased CDK2-AP1 (p12(DOC-1)) expression in human colorectal cancer (CRC) cell lines. In those same studies, induction of CDK2-AP1 expression promoted both cell cycle arrest and apoptosis. The goals of our present study were to better understand the mechanisms leading to reduced CDK2-AP1 expression in microsatellite unstable (MSI) CRC and to study further the effect of CDK2-AP1 modulation on cell proliferation and apoptosis utilizing RNA interference (RNAi) techniques. We used direct sequencing to screen for mutations of the poly (T)8 microsatellite-like region in the 3' end of the CDK2-AP1 gene in 24 CRC cell lines. We then utilized an in vitro human mismatch repair (MMR) recombinant system to assess for correction of the mutation and changes in CDK2-AP1 expression secondary to hMLH1 transfection. We also investigated the effect of CDK2-AP1 modulation in four settings: (1) native CDK2-AP1 absence, (2) endogenous CDK2-AP1 expression, (3) RNAi-induced CDK2-AP1 inhibition and (4) induced CDK2 AP1 over expression. The mutation - del T poly (T)8 - at the 3' end of the CDK2 AP1 gene was found in 3/12 (25%) of MSI CRC cell lines, but in none of the microsatellite stable samples (0/12). Interestingly, when wild-type MMR protein - MLH1 - was induced in an in vitro human recombinant system, the del T poly (T)8 mutation was reversed and CDK2-AP1 expression increased. RNAi-mediated CDK2-AP1 inhibition was associated with decreased apoptosis and increased cell proliferation in CDK2-AP1-non deficient CRC cell lines. We conclude that mutations in the microsatellite-like sequence of the CDK2-AP1 gene in MSI CRC are associated with decreased CDK2-AP1 expression. In addition, modulation of CDK2 AP1 expression in human CRC alters cell proliferation and apoptosis. PMID- 15806177 TI - Ethnic differences in mental health and lifestyle issues: results from multi-item general practice screening. AB - AIM: To determine ethnic differences in response, acceptance and desire to address problems identified by the multi-item screening tool (MIST). METHODS: Patients were assessed using the MIST in urban Auckland. Fifty consecutive adult patients from 20 randomly selected practices completed the MIST and evaluation sheet before their consultation. All patients and general practitioners (GPs) completed feedback forms. Analysis adjusted for the clustered nature of the data. RESULTS: Participants were 1000 patients and 20 urban GPs. The participation rate was 87% of GPs and 97.75% of patients. Compared with New Zealand Europeans, Pacific Island people were significantly more likely to be concerned about abuse and anger control. Maori were significantly more likely to want help with cutting down their alcohol use. The screening tool was accepted by all patients (<1% objection rate) regardless of ethnicity. DISCUSSION: This is the first study to examine possible ethnic differences between primary care patients' response to screening on lifestyle behaviours and mental health issues. PMID- 15806178 TI - SIT (seated immobility thromboembolism) syndrome: a 21st century lifestyle hazard. AB - The association between prolonged seated immobility and venous thromboembolism (VTE) is well recognised in relation to travel. In this case series, we report the occurrence of deep vein thrombosis (DVT) or pulmonary embolism (PE) following prolonged sitting in relation to work and/or recreation. The cases were characterised by the considerable length of time the individuals sat, the extensive nature of the VTE events in young individuals without other recognised risk factors, and the lack of recognition by the attending doctors of seated immobility as the likely major risk factor. While recognising that the association may be coincidental rather than causal, we propose the acronym 'SIT' (seated immobility thromboembolism) syndrome to encompass all forms of seated immobility associated with VTE. PMID- 15806179 TI - Heathrow to ICU direct. AB - We present the case of a witnessed massive pulmonary embolism (PE) resulting in pulseless electrical activity (PEA) following a long-haul flight. Our patient was successfully treated with cardiopulmonary resuscitation (CPR) and thrombolysis in the Emergency Department at Christchurch Hospital and went on to complete her honeymoon in New Zealand. PMID- 15806180 TI - Pulmonary embolus successfully treated with thrombolysis in the resuscitation room. PMID- 15806181 TI - Venous thromboembolism: when inaction can be fatal. PMID- 15806182 TI - Few rural general practitioners use the Internet frequently in regard to patient care. AB - AIMS: To benchmark North Island rural general practitioner (GP) access to computers and the Internet, both at work and home, and assess whether rural GPs are using the Internet in regard to patient care. METHODS: Cross-sectional postal survey of all North Island rural GPs in mid-2003. RESULTS: 175 of 289 GPs (60.6%) returned useable questionnaires. Most (89.0%) reported computer availability at work when consulting, but even more had access to a computer at home (97.1%, p<0.01). Access to the Internet was also lower at work (68.6%) than at home (98.8%, p<0.01). Fewer GPs (p<0.05) reported ever using the Internet at work in regard to patients (56.5%) than at home (71.9%). Less than 10% of all GPs used the Internet three or more times a week at work (6.9%) or home (8.6%) in regard to patients. Of those with Internet access at work, 27.0% had broadband (fast Internet) access. Predictors of having (versus not having) work Internet access were computer availability in consultations (p=0.04). CONCLUSIONS: Few North Island rural GPs use the Internet frequently in regard to patient care, despite increasing access to computers and the Internet, both at work and home. PMID- 15806183 TI - Patterns of medical practice variation: variability in referral for back pain by New Zealand general practitioners. AB - AIMS: To describe patterns of variation in referral among general practitioners, and to establish whether variability among practitioners within a geographic area is associated with high levels of utilisation in an area. METHOD: Multilevel analysis of routinely collected primary care data. The four outcome measures were referral to physiotherapy, specialist assessment, radiology, and approval of earnings-related compensation. RESULTS: The pattern of observed variability is not consistent for different referral activities: groups of general practitioners within one area may practice consistently in referral for one outcome, but be highly variable for another, while practitioners in other areas can show the reverse pattern. The degree of variability among GPs within geographic areas was not significantly correlated at the 95% level with the absolute level of referral to any of the referral options. CONCLUSION: The mechanisms which drive variability operate at the level of the specific clinical management option, rather than at the level of the overall approach to management of the disease. Caution should be exercised about claims that reductions in variability will produce reductions in utilisation. PMID- 15806184 TI - Documentation of family violence in New Zealand general practice. AB - AIM: To determine the rate of family violence documented during general practice consultations and describe clinical presentations. METHOD: A dataset of 447,809 computerised consultations involving 143,634 patients from 41 general practices throughout New Zealand was examined to identify consultations recording family violence issues. The documentation rate was determined and the subset analysed. RESULTS: A subset of 337 consultations from the 447,809 examined (0.075%) involved a family violence issue. This subset included 311 patients, 0.2% of the 143,634 patients in the original 6-month dataset. 225 (81%) of the patients in the subset were female. Family violence was the main reason for presentation in 137 (40%) consultations. The perpetrator was identified as the partner in 134 (40%) consultations, as the parent in 54 (16%) consultations and the patient identified themselves as the possible abuser in 17 (5%) consultations. Physical abuse (42%) and sexual abuse (26%) was most commonly mentioned. Past abuse (42%) was discussed as often as current abuse (41%). Depression and anxiety disorders were documented in 59 (18%) of these consultations. CONCLUSIONS: The number of consultations documenting family violence is low in this dataset. Such information is not always recorded, however GPs can also be reluctant to ask about, and patients can be hesitant to disclose family violence issues. The number of consultations involving the perpetrator was higher than expected. GPs require training to deal with both the victim and the perpetrator of family violence. PMID- 15806185 TI - Responsible IVF treatment in New Zealand is the preferential transfer of a single embryo. PMID- 15806186 TI - Reflecting on the 'difficult' patient. PMID- 15806187 TI - Screening for prostate cancer: a response to four letters. PMID- 15806188 TI - Screening for prostate cancer: the view of the Prostate Awareness and Support Society's President. PMID- 15806189 TI - Regarding 'Informing consent in New Zealand research: researchers' conflict of interest and patient vulnerability'. PMID- 15806190 TI - Modern surveillance needed to control the increase in STIs. PMID- 15806191 TI - COX-2 inhibitors--second, do some good. PMID- 15806192 TI - The epidemiology of multiple sclerosis in New Zealand. PMID- 15806193 TI - Are DEXA results reproducible? PMID- 15806195 TI - Marine natural products: synthetic aspects. AB - An overview of marine natural products synthesis during 2003 is provided. The emphasis on total syntheses of molecules of contemporary interest, new total syntheses, and syntheses that have resulted in structure conformation or stereochemical assignments. PMID- 15806196 TI - Natural products to drugs: natural product derived compounds in clinical trials. AB - Natural product and natural product-derived compounds that are being evaluated in clinical trials or in registration (current 31 December 2004) have been reviewed. Natural product derived drugs launched in the United States of America, Europe and Japan since 1998 and new natural product templates discovered since 1990 are discussed. PMID- 15806197 TI - Muscarine, imidazole, oxazole and thiazole alkaloids. AB - Novel and structurally diverse natural products containing imidazol-, oxazole-, or thiazole-unit(s) display a wide variety of biological activities. The isolation, biological activity and total synthesis of naturally occurring muscarine, imidazole, oxazole and thiazole alkaloids have been reviewed. The literature covers from January 2003 to June 2004. PMID- 15806198 TI - Triterpenoids. AB - This review covers the isolation and structure determination of triterpenoids including squalene derivatives, lanostanes, cycloartanes, dammaranes, euphanes, tirucallanes, tetranortriterpenoids, quassinoids, lupanes, oleannes, friedelanes, ursanes, hopanes, isomalabaricanes and saponins. The literature from January to December 2002 is reviewed and 221 references are used. PMID- 15806199 TI - beta-Phenylethylamines and the isoquinoline alkaloids. AB - This review covers beta-phenylethylamines and isoquinoline alkaloids derived from them, including further products of oxidation, condensation with formaldehyde and rearrangement, some of which do not contain as isoquinoline system, together with napthylisoquinoline alkaloids, which have a different biogenetic origin. The occurrence of the alkaloids with the structures of new bases, together with their reactions and syntheses, are reported. The literature from July 2003 to June 2004 is reviewed, with 145 references cited. PMID- 15806200 TI - Acetogenins from Annonaceae: recent progress in isolation, synthesis and mechanisms of action. AB - The aim of the present review is to summarise the knowledge about newly isolated acetogenins (ACGs) in the last six years. It will also report the total syntheses that have allowed either the confirmation or the revision of some structures, together with the biological activities and mechanism of action of such interesting natural products. In fact, of the 417 isolated compounds reviewed, over 176 have been added during the period from 1998 to 2004. PMID- 15806201 TI - The epidemic of the metabolic syndrome. AB - The incidence of metabolic syndrome (MS) is rising worldwide. This is partly due to a significant increase in the prevalence of obesity. Observational cross sectional studies as well as demographic health surveys from the Middle East, point out that the prevalence of obesity increases from an average of 6% in healthy children to 20% in adolescent males and to a further 32% in elderly patients. The impact of obesity on our population is expected to be considerable; especially, as it feeds into further rising in the prevalence of hypertension, diabetes, MS and cardiovascular disease. The prevalence of MS in nondiabetic adults in Europe was recently reported to be 15%. In the Middle East, as pointed out by pilot observational projects, is estimated to be anywhere between 15-25%. The medical system is unprepared to deal with this epidemic partly due to scanty knowledge on the clinical significance of the MS and importantly as there is a limited number of specific treatments that we can offer these patients. PMID- 15806202 TI - Female genital mutilation and childbirth. AB - The practice of female genital mutilation sadly remains to this day, part of the life of many women worldwide. In the Middle East and Africa, it is hard to practice obstetrics and gynecology without some knowledge of the condition. This review is an attempt to discuss the salient points as regards to management of these women during pregnancy and labor. PMID- 15806203 TI - Chromosomal instability detected by interphase fluorescence in situ hybridization and its relation to p53 alteration in prostate carcinoma in Saudi patients. AB - OBJECTIVE: Chromosomal instability (CIN) is a feature of human neoplasm. The p53 mutation has been shown to be associated with CIN in many human dysplastic and neoplastic lesions. The objective of this study was to examine CIN and p53 mutations in prostate carcinoma (Pca) resected from Saudi patients. METHODS: Testing of p53 alteration using immunohistochemistry was performed on 28 archived prostatic carcinoma specimens containing Pca foci from Saudi patients seen at King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia. Chromosomal instability was evaluated in the same tissues by interphase in situ hybridization (IFISH) using centromere probes for chromosome 7 and 8. Immunohistochemistry and IFISH were performed at Princess Margaret Hospital, University Health Network, Toronto, Ontario, Canada in 2001. RESULTS: The p53 immunoreactivity was found in 29% in Pca and 0% in benign epithelium. Interphase in situ hybridization revealed numerical chromosomal alterations in keeping with CIN in 63% of p53 positive and 20% p53 negative Pca. No evidence of CIN was seen in non-neoplastic epithelium. CONCLUSION: We concluded that CIN as determined by IFISH is present in Pca from Saudi patients similarly to those reported in western countries. The p53 mutation occurs relatively infrequently in Pca and is associated with the presence of CIN at least in a subset of Pca. PMID- 15806204 TI - Immunohistochemical analysis of CD71, CD98 and CD99 activation antigens in human palatine and nasopharyngeal tonsils. AB - OBJECTIVE: Tonsils (palatine and nasopharyngeal) are immunologically active tissues. Due to their anatomical location, they are considered to be the initial defense barrier against the antigens entering into the respiratory and gastrointestinal tract. Tonsils act against these antigens by producing and activating the lymphocytes, which are responsible for the immune response. In order to get information regarding the distribution of cell surface antigens on the epithelial, stromal and lymphoid cells of these organs, we performed immunohistochemical staining by using antibodies against CD99, CD71 and CD98 activation antigens. METHODS: Tissue samples of 20 patients undergoing tonsillectomy and adenoidectomy who presented with recurrent tonsillitis and adenoid hypertrophy in the Otorhinolaryngology Department, Hacettepe University Medical Faculty Hospital, Ankara, Turkey in 2001, were obtained as partial tissue samples apart from pathological examination. Tissues were immunostained by the indirect immunoperoxidase method. RESULTS: Strong CD71 reactivity in macrophages was observed as an indicator of the active role of the macrophages in immunoresponse in the chronic inflammation reaction. The CD98 reactivity on the proliferative basal layer of epithelium was a usual finding, as its detection in epithelial neoplasms and proliferative states is well known. We did not observe any reactivity of CD98 in nasopharyngeal tonsil epithelium and lymphoid cells of either nasopharyngeal or palatine tonsils. The CD99 reactivity was observed in the T-cell dependent area. CONCLUSION: We determined some topographic difference in the expression of some activation antigens in the epithelial, stromal and lymphoid components of the palatine and nasopharyngeal tonsils. Further detailed studies directed to determine the role of these antigens in tonsils would help to understand the role of these molecules in inflammatory events. PMID- 15806205 TI - Comparison of light cycler PCR and conventional susceptibility testing for detection of MRSA from cultures. AB - OBJECTIVE: To compare a duplex light cycler polymerase chain reaction (PCR) assay targeting the mecA gene and a Staphylococcus aureus (S. aureus) specific marker and the conventional method. METHODS: We evaluated 400 samples sent to the laboratory in Zayed Military Hospital, Abu Dhabi, United Arab Emirates for methicillin-resistant Staphylococcus aureus (MRSA) screening and routine bacterial cultures from the period January 2003 to January 2004. All samples were cultured and identified according to the National Committee for Clinical Laboratory Standard guidelines. Staphylococcus aureus were tested for methicillin susceptibility according to the guidelines. All Staphylococcus positive cultures underwent testing by the new duplex light cycler PCR assay. We used 2 pairs of primers: mecA and nuc. Both targeted the mecA gene and the S. aureus-specific marker. Results obtained from the 2 methods (conventional culture method and the real-time PCR method) were compared. RESULTS: From the 400 samples tested, a total of 9 MRSA were detected by both methods. The real-time PCR method took less than 60 minutes to complete. CONCLUSION: This study shows that the duplex light cycler PCR assay method is very sensitive, very specific, and less time consuming in diagnosing MRSA from bacterial cultures. PMID- 15806206 TI - Azithromycin prophylaxis and treatment of murine toxoplasmosis. AB - OBJECTIVE: To evaluate the azithromycin effects alone and in combination with other agents in the prophylaxis and treatment of murine toxoplasmosis. METHODS: A total of 280 BALB/c mice were included, and 2 x 103 Toxoplasma organisms of the RH strain Toxoplasma gondii strain ATCC50174 were given intraperitoneally to each mouse. In experiment one, 40 animals were given azithromycin 200 milligram/kilogram/daily for 3 days starting the day of inoculation, 40 mice were control. In experiment 2, the treatment was started 48 hours after inoculation and given daily for 3 days: one group received azithromycin 200 milligram/kilogram/day, the second group received pyrimethamine 25 milligram/kilogram/day, and the sulfadiazine 100 milligram/kilogram/day. The third group was control. In experiment 3, 7 groups of animals received one of the following (1) none, (2) azithromycin 200 milligram/kilogram/day, (3) pyrimethamine 25 milligram/kilogram/day and sulfadiazine 100 milligram/kilogram/day, (4) azithromycin and sulfadiazine, (5) azithromycin and pyrimethamine, (6) azithromycin with sulfadiazine and pyrimethamine, (7) sulfadiazine alone. Treatment was initiated 72 hours after inoculation for 3 days. The study was conducted at the Animal Care Facility of King Saud University, Riyadh, Kingdom of Saudi Arabia. RESULTS: Animals that received azithromycin simultaneously with inoculation survived, and all control animals died. All animals died in groups receiving single drug therapy. Animals treated with azithromycin and sulfadiazine showed a survival rate of 40%, sulfadiazine and pyrimethamine 40%, or azithromycin with sulfadiazine and pyrimethamine 95% (p<0.0001). CONCLUSION: Azithromycin alone was found to be effective in the prophylaxis of murine toxoplasmosis. Combination therapy was effective in the treatment of murine toxoplasmosis. PMID- 15806207 TI - Effect of cholesterol diet on reproductive function in male albino rats. AB - OBJECTIVE: The present study was carried out to investigate the effect of cholesterol diet (400 mg/kg body weight) for 60 days on gonadal function in albino rats. METHODS: The study was conducted in the Animal House Unit at Jordan University of Science and Technology, School of Medicine, Irbid, Jordan between October 2003 and February 2004. Adult male and female albino rats of Sprague Dawley strain were raised under controlled temperature and light. Male rats were divided into: a) control group--rats receiving vehicle (olive oil) for 60 days and treatment group--rats receiving cholesterol diet for a reproductive cycle. Animals were weighed and autopsied 24 hours after the last dose. Biochemical and histological approaches were used to assess fertility in both groups. RESULTS: The treatment caused significant reduction (p < or = 0.001) in sperm motility and density in cauda epididymides and testes. A significant reduction (p < or = 0.001) in epithelial cell height of caput, cauda and seminal vesicle was also observed. In the treated group, there was a significant reduction (p<0.001) in seminiferous tubules diameter and Leydig cell nuclear diameter. Spermatocytes (primary and secondary) were significantly decreased (p < or = 0.01) and spermatids were significantly reduced (p < or = 0.001) in the treatment group. Whereas, the number of degenerating Leydig cells (interstitial cells) increased significantly (p < or = 0.001). Serum biochemistry reveals significant increase (p<0.001) in cholesterol and triglyceride levels. The intragastric administration of cholesterol diet to male rats for 60 days significantly reduced the number of females impregnated by these males. However, the number of implantations and number of viable fetuses were significantly (p<0.01) decreased in female rats impregnated by males that ingested cholesterol. On the other hand, the number of resorptions was significantly (p<0.01) increased in females impregnated by males that ingested cholesterol. The histometry and histology of reproductive organs confirm these results. CONCLUSION: Hyperlipidemia can cause alteration in the biochemistry and histometry of reproductive organs and can cause inhibition of spermatogenesis via the Leydig cell. PMID- 15806208 TI - Biological and morphological effects on the reproductive organ of rats after exposure to electromagnetic field. AB - OBJECTIVE: The biological effect of electromagnetic field (EMF) emitted from mobile phones is a current debate and still a controversial issue. Therefore, little is known on the possible adverse effects on reproduction as mobile phone bio-effects are only a very recent concern. The aim of this experimental study was to determine the biological and morphological effects of 900 MHz radiofrequency (RF) EMF on rat testes. METHODS: The study was performed in the Physiology and Histology Research Laboratories of Suleyman Demirel University, Faculty of Medicine, Isparta, Turkey in May 2004. Twenty adult male Sprague Dawley rats weighing 270-320 gm were randomized into 2 groups of 10 animals: Group I (control group) was not exposed to EMF and Group II (EMF group) was exposed to 30 minutes per day, 5 days a week for 4 weeks to 900 MHz EMF. Testes tissues were submitted for histologic and morphologic examination. Testicular biopsy score count and the percentage of interstitial tissue to the entire testicular tissue were registered. Serum testosterone, plasma luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels were assayed biochemically. RESULTS: The weight of testes, testicular biopsy score count and the percentage of interstitial tissue to the entire testicular tissue were not significantly different in EMF group compared to the control group. However, the diameter of the seminiferous tubules and the mean height of the germinal epithelium were significantly decreased in EMF group (p<0.05). There was a significant decrease in serum total testosterone level in EMF group (p<0.05). Therefore, there was an insignificant decrease in plasma LH and FSH levels in EMF group compared to the control group (p>0.05). CONCLUSION: The biological and morphological effects resulting from 900 MHz RF EMF exposure lends no support to suggestions of adverse effect on spermatogenesis, and on germinal epithelium. Therefore, testicular morphologic alterations may possibly be due to hormonal changes. PMID- 15806209 TI - Intracytoplasmic sperm injection after total conventional in-vitro fertilization failure. AB - OBJECTIVE: This study assessed the value of delayed intracytoplasmic sperm injection after failure of initial in-vitro fertilization in patients with normal seminology. METHODS: A case series analysis was conducted on 82 couples who underwent delayed intracytoplasmic sperm injection after failure of initial in vitro fertilization at Fakeeh Hospital, Jeddah, Kingdom of Saudi Arabia between January 1995 and January 2001. RESULTS: Seventy-eight couples had 616 unfertilized metaphase II mature oocytes after initial in-vitro fertilization. Second day "rescue" reinsemination by intracytoplasmic sperm injection (ICSI) resulted in 174 embryos that were suitable for transfer. This resulted in 4 clinical pregnancies (5.1% per started rescue ICSI cycle). CONCLUSION: The limited success rate in this study provides confirmatory data that rescue ICSI is not an efficient adjuvant laboratory procedure. PMID- 15806210 TI - Occupational exposure to urban pollutants and plasma insulin. AB - OBJECTIVE: The aim of this study was to investigate whether traffic police of a big city who are exposed to urban pollutants are at risk to alterations in plasma insulin concentrations compared with controls. METHODS: The class of workers examined in this study were employees of the municipal police in Rome, Italy. The study was carried out during the period March-April 2001. Traffic police were matched with a control group, where gender, age, working life and drinking habit were noted. A total of 246 traffic police (128 men and 118 women) with outdoor activity and exposure to urban pollutants and 242 subjects (128 men and 114 women) with indoor activity were included in the study. RESULTS: The levels of plasma insulin were significantly lower in traffic police compared to the control group of both gender. CONCLUSION: Plasma insulin level was altered in traffic police who are exposed to chemical and physical stressors. PMID- 15806211 TI - Comparison of the efficacy of inhaled budesonide and oral choline in patients with allergic rhinitis. AB - OBJECTIVE: A single blind parallel group study was conducted to evaluate the effects of oral choline [given as tricholine citrate (TRI)] in patients with allergic rhinitis, and compare its efficacy with intranasal budesonide (BUD). METHOD: The study was conducted at the Department of Respiratory Medicine, Vallabhbhai Patel Chest Institute, Delhi, India from February 2001 to April 2002. Sixty patients were randomized into 2 groups after a run-in period of 2 weeks. Group A received intranasal BUD 200 microg twice daily and group B received TRI 500 mg thrice daily. The patients were reviewed every 2 weeks up to 8 weeks. The mean individual symptom score, total symptom score and drug score were significantly reduced in both groups (p<0.05) compared to baseline values, with maximum effect occurring within 4 weeks of therapy. RESULTS: Budesonide showed statistically significant reduction (p<0.05) in all the outcome parameters, when compared to TRI. Crossover study between the 2 treatment groups also showed similar results. Seventy-six percent of patients with BUD and 43% of patients with TRI found the drug to be effective. CONCLUSION: Both intranasal BUD and oral TRI are effective in relieving symptoms of allergic rhinitis. Budesonide was found to be the statistically superior drug. PMID- 15806212 TI - Age-related changes in human tendo calcaneus collagen fibrils. AB - OBJECTIVE: The ruptures of tendo calcaneus often occur between the age of 30-45 years as described by several textbooks. It is also described that some diseases and drugs are said to be responsible in the etiology; however, there are no studies related with the detailed histological structure of collagen fibrils found in the tendon in the age groups of humans. In view thereof, this study was aimed to obtain further information on the etiology and to find an answer regarding the frequency of the ruptures occurring between the age of 30-45 years in humans. METHODS: In the study, the biopsy specimens taken from 28 patients (ages 1-68 year) who had undergone surgery due to tendo calcaneus ruptures or achilloplasty operation were examined by transmission electron microscope. All the specimens were prepared according to routine electron microscopic tissue preparation technique. The patients were divided into 7 age groups (1-9, 10-19, 20-29, 30-39, 40-49, 50-59, >60 years) and there were 4 patients in each group. The transverse diameters of collagen fibers were measured from the ultra thin sections and statistical analysis of the results were performed. The study was carried out in the electron microscopy laboratory of the Anatomy Department of Hacettepe University, Ankara, Turkey between January 2004 and September 2004. RESULTS: The diameters of the collagen fibers were higher in the 20-29 year-old group compared to other groups and it showed a statistically significant difference. In patients who were in the 30-39 year-old group or older, the diameters of the collagen fibers were lesser than the 20-29 year-old group. However, an increase was observed in the collagen fibril concentration of these groups. In examination of the specimens of patients who were under 20-year-old, the diameter of the collagen fibers were less than the 20-29 year-old group. The electron microscopic appearance of the tissue sample of a one-year-old patient had a specific organization and in this patient, both the diameters and concentration of collagen fibers were less. CONCLUSION: We believe that the decrease in the diameters of collagen fibers of 30-45 year-old patients who are in the active period of their life, can play a role in the etiology of the frequency of tendo calcaneus ruptures similar to other etiologic factors. PMID- 15806213 TI - Immunohistochemical evaluation of periprosthetic membrane from loose cemented and uncemented total hip arthroplasty. AB - OBJECTIVE: Evaluate the cellular mechanisms responsible for the aseptic loosening of total hip replacements. METHODS: Twenty periprosthetic membranes were collected during revision procedures at the Department of Orthopedics, Hacettepe University, Ankara, Turkey, during 1998 to 1999. Arthroplasties with polyethylene components and without polyethylene components were examined under light microscopy and immunohistochemically using the indirect immunoperoxidase technique into 2 different groups. RESULTS: Immunohistochemical analysis with monoclonal antibodies reactive to certain cell surface antigens (CD45, CD44, CD98, CD31, CD26, CD71) revealed different morphological characteristics regarding the adjacent fibrosis, leukocyte activation, and immunologic response. CONCLUSION: These findings suggested the role of different cellular mechanisms in each group. PMID- 15806214 TI - Carcinoid tumors of the appendix. Our experience in a university hospital. AB - OBJECTIVE: To present our experience of carcinoid tumors of the appendix managed at a university teaching hospital. Complex symptomatology, varied biochemical affections and different surgical therapeutic modalities are discussed. METHODS: The medical records of all the patients who underwent consecutive appendectomies at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia from 1994 to 1999 were retrospectively analyzed. The data of patients identified to have histological evidence of carcinoid tumors of the appendix were further reviewed for the demographic details, indications for surgery, surgical procedure, tumor localization in the appendix and size; concomitant appendicitis and further surgical procedures were considered. RESULTS: During the study period, 1547 appendectomies were performed and, out of these, 9 (0.6%) cases were reported to have carcinoid tumors of the appendix. There were 4 male and 5 female patients, age range 17-51 years (median 29.8 years). Seven subjects had a clinical evidence of appendicitis while 2 presented with chronic abdominal pain. There were 6 open and 3 laparoscopic appendectomies. Six carcinoid tumors were encountered at the appendiceal apex, 2 at the midportion, and one at the base with a mean diameter of 9.5 mm (range, 4-19 mm). One patient had histologically confirmed residual tumor, which necessitated a right hemicolectomy 3 weeks later. All patients remained disease-free during a mean follow up of 7 years (range, 4-10 years). CONCLUSION: Carcinoid tumors of the appendix are extremely rare and invariably remain asymptomatic. Simple appendectomy offers adequate relief while the need for further extensive surgery depends on tumor characteristics and dissemination. Despite an excellent prognosis, all reported patients should be followed up with urinary 5-hydroxyindoleacetic acid and abdominal ultrasonography. PMID- 15806215 TI - Multinodular goiter management in Western Saudi Arabia. AB - OBJECTIVE: The aim of this study was to provide epidemiological data and summarize the different modalities of management of multinodular goiters (MNG) in the Western Province of Saudi Arabia (KSA). METHODS: A total of 135 patients had MNG, which were diagnosed by ultrasonography at King Abdul-Aziz University Hospital in Jeddah, KSA between January 2003 and June 2004. The data collected includes the age, gender, physical examination of the thyroid gland (trifluorothymidine, free thyroxine, free triiodothyronine, thyroid stimulating hormone), ultrasonographic findings and Tc99 radioiodine nuclear scan of the thyroid gland, fine needle aspiration (FNA) of the gland and antithyroid antibodies. The study group was divided according to the thyroid status: nontoxic euthyroid MNG, hypothyroid MNG and toxic hyperthyroid MNG. The management of MNGs according to the patients' clinical presentations and thyroid status was reviewed. RESULTS: The mean age was 39 +/- 12.66 years with a range of 10-79 years. Forty-two patients (31%), with no history of thyroid cancer, had ultrasoundguided FNA; 41 (97.6%) of the FNAs were benign. One FNA (2.3%) was positive for papillary carcinoma in a 56-year-old female patient. Thyroid antibodies (anti-microsomal antibodies and anti-thymoglobulin antibodies) were measured in 50 patients (37%). Thirty-one (62%) were positive, 11 (35%) of which were positive in patients with documented hypothyroidism. All patients with hypothyroidism were treated with levothyroxine; however, only 25.6% with euthyroid MNG were treated with suppressive doses of levothyroxine. Twelve patients (44.4%) with toxic MNG were treated with antithyroid medications. Radioiodine therapy was not given to any patient with nontoxic MNG; however, 48% of those with hyperthyroid MNG received radioactive iodine treatment. Surgery was carried out in 25.6% of patients with euthyroid nontoxic MNG, and in 11 patients with toxic MNG. Nearly half of those with nontoxic MNG (46.5%) and 14.8% of those with a subclinical hyperthyroid MNG refrained from any therapy. They were followed-up by their clinician in the outpatient clinic. CONCLUSION: Serum thyroid stimulating hormone levels, ultrasound and fine needle biopsy were the cornerstones of the diagnostic evaluation of patients with MNG. Review of the management of these patients was comparable to that found in the literature. The treatment strategy was similar to the recommendations by the American Thyroid Society; however, radioactive iodine treatment was not used as a treatment for patients with nontoxic goiters. PMID- 15806216 TI - Influence of dietary oils on liver and blood lipid peroxidation. AB - OBJECTIVE: Diets high in unsaturated fatty acids have been recommended to lower the risk of cardiovascular disease. However, these lipids are more susceptible to lipid peroxidation than saturated fatty acids. The aim of the present study described herein was to investigate the effects of dietary oils (differing in their degree of saturated and unsaturated fatty acids) on liver and blood lipid peroxidation in chicks. METHODS: The experiments were conducted at the laboratories of University of Dumlupinar, Kutahya, Turkey and Osmangazi University, Eskisehir, Turkey between November 2002 and December 2003. The animals were randomly divided into 5 groups of 30 and fed dietary butter, margarine, olive oil, sunflower oil or corn oil for 7 weeks. Liver malondialdehyde level, blood superoxide dismutase activity (SOD) and glutathione peroxidase activity (GPx), serum vitamin E, and total antioxidant (AOA) levels were measured to determine the effects of the dietary oils on lipid peroxidation. RESULTS: No significant differences were observed in SOD and GPx activities, or vitamin E and AOA levels between the experimental groups. However, the results indicated that the corn oil feeding caused significant increases in liver malondialdehyde (a genotoxic byproduct of lipid peroxidation) level as compared with the other oils. CONCLUSION: The results demonstrate that corn oil feeding increases lipid peroxidation significantly and thus may raise the susceptibility of tissues to free radical oxidative damage. PMID- 15806217 TI - Predictors of gangrenous necrotizing enterocolitis and extent of disease. Early laparotomy versus peritoneal drainage. AB - OBJECTIVE: Evaluation of known predictors of gangrene in neonates with necrotizing enterocolitis (NEC) and identification of those suggestive of severe disease, requiring expeditious laparotomy rather than primary peritoneal drainage as a definitive treatment. METHODS: This is a retrospective review of data collected from the medical records of newborns with confirmed NEC, treated at the Maternity and Children's Hospital, Dammam, Kingdom of Saudi Arabia, from May 1993 to May 2004. Fifty-five cases were selected for the study, 23 had successful medical management and 32 underwent laparotomy. Of this group, 15 had peritoneal drainage prior to laparotomy. Nine known clinical, radiological and laboratory features suspicious of bowel perforation or gangrene were evaluated. The operated group was classified according to the extent of disease into isolated, multifocal or pan intestinal and the distribution of these 9 criteria was calculated for each of the 3 groups. Comparison was then carried out between the group with isolated NEC and those with extensive disease. RESULTS: Isolated NEC was present in 8 (25%), multifocal NEC in 19 (59%) and pan intestinal NEC in 5 (16%) of the operated cases. Pneumoperitoneum and palpable abdominal mass were the most specific and predictive signs of perforated or gangrenous bowel in NEC. Severe pneumatosis intestinalis and gasless abdomen were also highly specific and predictive of the same but had a low prevalence. Abdominal wall erythema, persistent metabolic acidosis, portal vein air, gasless abdomen and severe pneumatosis intestinalis were found to be associated with severe or extensive gangrene. Palpable abdominal mass and fixed dilated loops were increased in cases of isolated NEC. Portal vein air was associated with the highest mortality. CONCLUSION: Pneumoperitoneum, though the only absolute evidence of bowel perforation, cannot predict the extent of disease. Peritoneal drainage is a useful stabilizing procedure but the presence of any of the above mentioned criteria which are associated with severe disease necessitate a quick decision in favor of laparotomy. The absence of these signs, however, cannot rule out extensive or progressive NEC and failure to improve after peritoneal drainage also requires an emergency laparotomy, regardless of birth weight or gestational age. PMID- 15806218 TI - Head size at birth in neonates with transposition of great arteries and hypoplastic left heart syndrome. AB - OBJECTIVE: Appropriate fetal brain growth depends upon the cerebral blood flow (CBF). Different congenital heart defects (CHDs), due to the difference in anatomy and physiology, alter the intrauterine CBF. Thus, variable brain growth is expected in different CHDs that is reflected by variability in the head circumference (HC) at birth. The present study was carried out to compare the HC of babies born with transposition of great arteries (TGA) and hypoplastic left heart syndrome (HLHS) in comparison to normal control. METHODS: The data on the HC of neonates with TGA and HLHS were extracted from the computer database then compared with the control group. During the period from January 1996 to December 2003, a total of 7396 neonates were admitted, out of which 639 (8.6%) were admitted with the diagnosis of the CHD. After correcting for gestational age and non-availability of HC measurements, 236 infants were excluded. Out of the remaining 403 term appropriate for gestational age (AGA) infants, 46 had TGA while 28 had HLHS. The control group comprised of a total of 74 term AGA infants. RESULTS: The mean HC for the control group was noted to be 34.4 +/- 1.7 centimeters (cms), 33.7 +/- 1.5 cms for TGA while it was 32.9 +/- 1.3 cms for HLHS. The head size at birth for newborn with HLHS was significantly smaller than the TGA (p=0.03) and control group (p=0.001). Similarly, HC of TGA group was significantly smaller than the control group (p=0.02). CONCLUSION: The newborns with TGA and HLHS are found to have significantly small head size at birth. The clinical significance of this finding with respect to the neurodevelopmental outcome and value of early antenatal intervention to repair these defects remains to be evaluated in further studies. PMID- 15806219 TI - Prevalence of hypocalcemia in children examined for serum calcium in Sana'a, Yemen. AB - OBJECTIVE: To estimate the prevalence of hypocalcemia in children examined for serum calcium. METHODS: A record-based study was carried out in the Specialized Pediatric Center in Sana'a city, Yemen during the 4 year period 1999 to 2003. Out of 90600 patients seen for different causes, 310 (0.34%) were subjected to serum calcium level examination. Data regarding age and gender were also collected. RESULTS: The median age of the patients was 5 months, with minimum age of one day and maximum age of 4 years. Prevalence of hypocalcemia among children examined for serum calcium was 58%, 60% of them were males and 40% were females. Age group of 0-1 months constituted 17.8% while the age group of 1-3 months constituted 17.2%. Patients with serum calcium level between 4-6 mg constituted 27.8% and less than 7.5 mg were 72.2%. CONCLUSION: Prevalence of hypocalcemia in children examined for serum calcium was high. Males were more affected than females and was more in neonates followed by age 1-3 months then age 3-6 months. PMID- 15806221 TI - Schatzki's ring. An unusual clinical and radiological presentation. AB - Schatzki's ring is a lower esophageal mucosal ring associated with a small sliding hiatus hernia. Most investigators described it as either an asymptomatic or symptomatic entity with chronic recurrent presentation of dysphagia. Barium swallow study in patients with Schatzki's ring was described as a thin smooth circumferential constriction at the GE junction. This case report describes an unusual clinical and radiological presentation in a patient with Schatzki's ring. Our adult male patient experienced sudden dysphagia, followed by spontaneous relief after an interval of 12 hours without specific treatment. Radiological findings were highly suggestive of lower esophageal malignancy. However, this possibility could be excluded by upper endoscopy and histopathological examination of biopsies taken from the lesion. The condition was diagnosed as Schatzki's ring with unusual clinical and radiological presentation. PMID- 15806220 TI - Risk factors for asthma among primary school children in Baghdad, Iraq. AB - OBJECTIVE: Asthma is one of the most common chronic diseases of children. To fill the gap in data concerning this disease in Iraq, we investigated the socio demographic and other risk factors related to asthma occurrence among primary school children. METHODS: A case control study was conducted in Baghdad, Iraq among primary school children aged 6-12 years, for the period between October 2000 and June 2002. Six hundred and forty-four children with asthma were investigated with a control group of 1618 children without asthma. Well constructed standardized modified questionnaires of International Study of Asthma and Allergies in Childhood were completed by the parents of the chosen children. RESULTS: From the studied risk factors, the following were found as significant risk factors for asthma development: crowding rate of > or = 5 (odds ratio [OR]=1.65, 95% confidence interval [CI]=1.1 - 2.4), lower educational level of parents, prematurity (OR=1.61, 95% CI=1.003-2.59), low birth weight (OR=2.41, 95% CI=1.87-3.09), family history of asthma whether father (OR=3.86, 95% CI=2.54 5.87), or mother (OR=8.27, 95% CI=5.21-13.15) or sibling (OR=4.33, 95% CI=3.24 5.8) and environmental exposure to tobacco smoking during pregnancy or currently from both parents. On the contrary, our study failed to detect significant association for the following factors: gender, residency, type of birth, breast feeding and duration. CONCLUSION: Crowding, low parental education, prematurity, low birth weight, family history of asthma and smoking are significant risk factors for asthma development among our primary school children. Efforts must be concentrated for hygienic environment, good antenatal care and quitting smoking habits in order to overcome this health problem. PMID- 15806222 TI - Frontal sinus cementifying ossifying fibroma. AB - Cementifying ossifying fibroma is a fibro-osseous lesion which produces cementum. Cementifying ossifying fibroma is a mesodermal, non-odontogenic tumor of ectopic multipotential periodontal membrane blast cells. Although, it is a benign bone tumor, it has an aggressive destructive behavior in patients under 20 years of age with high tendency for recurrence. Cementifying ossifying fibroma can be recognized microscopically by the presence of cementicles. This is a case of frontal sinus cementifying ossifying fibroma presenting with proptosis in a 39 year-old male. PMID- 15806223 TI - Successful treatment of Brucella endocarditis with aortic root abscess. AB - Brucella endocarditis is a rare but fatal complication of Brucellosis, it causes destructive valvular lesions. The aortic valve is the most common affected site. We present a case of Brucella endocarditis with aortic root abscess, the patient received a prolonged combination of antibiotic therapy, and underwent aortic valve replacement. After one and a half years of follow up, the patient is still without signs of recurrence. The high mortality in Brucella endocarditis can be overcome by early diagnosis and aggressive therapy. PMID- 15806224 TI - Survival of 2 extreme preterm neonates with cardiac tamponade as a complication of percutaneous central venous catheterization. AB - We report 2 extreme preterm neonates who developed cardiac tamponade secondary to perforation of the myocardium by the percutaneous silastic central venous catheters, which were inserted for parental nutrition. Percutaneous pericardiocentesis was performed and pericardial effusion was aspirated, later proved by analysis to be total parental nutrition. The lines were removed and the patients successfully resuscitated and survived, both were sent home in good condition. PMID- 15806225 TI - Hepatocellular carcinoma directly invading the duodenum. AB - Recurrent gastrointestinal bleeding from hepatocellular carcinoma HCC invading the duodenum is very rare. We present a case of 50-year-old male who was admitted with a history of recurrent upper gastrointestinal tract UGIT bleeding, weight loss and anemia. The patient was known to have a chronic hepatitis C. Endoscopic examination showed grade-2 non-bleeding esophageal varices, and a large ulcerated duodenal mass partially obstructing the duodenal bulb outlet and causing recurrent UGIT bleeding. Pathological evaluation of the mass revealed HCC. PMID- 15806226 TI - Does prolonged semi-erection in prepubertal high flow priapism result in increased penile size? AB - High flow priapism is a rare pathology resulting mainly from trauma to the perineum leading to arterial-lacunar fistula. Management includes arterial embolization using absorbable material, as well as conservative approach. In this case, the effect of prolonged semi-erection in prepubertal high flow priapism on increased penile size is discussed. PMID- 15806227 TI - Cecal fistula after percutaneous nephrolithotomy ended by hemicolectomy. PMID- 15806228 TI - Detection of antibodies to the extractable nuclear antigens by enzyme linked immunosorbent assay. PMID- 15806229 TI - Antiphospholipid syndrome among Bahraini patients. PMID- 15806230 TI - Contribution study of visceral leishmaniasis in Syria. PMID- 15806231 TI - Vitiligo and human herpesvirus 6. Is there a relationship? PMID- 15806232 TI - Use of laryngeal mask airway for the care of rhinoplasty. PMID- 15806233 TI - Breathlessness and respiratory failure in myasthenia gravis patient. PMID- 15806234 TI - Preliminary results of muscle diseases prevalence in patients from Jordan. PMID- 15806235 TI - Pregnancy outcome following exposure to orlistat, ramipril, glimepiride in a woman with metabolic syndrome. PMID- 15806237 TI - Retropubic space hemorrhage. An unusual complication in cesarean section. PMID- 15806236 TI - Randomized trial of ceftriaxone prophylaxis in elective cesarean section. PMID- 15806238 TI - Condylomata acuminata in infants and young children. Topical podophyllin an effective therapy. PMID- 15806239 TI - Neutropenia. PMID- 15806240 TI - Effects of angiotensin II and leptin on blood pressure. PMID- 15806241 TI - [Not Available]. PMID- 15806242 TI - [Epidemiology and geography: the pathogenic complex of Max. Sorre]. AB - Some essential features of the pathogenic complex concept formulated by the French geographer Max. Sorre are presented, aiming at integrating epidemiological and geografic studies. PMID- 15806243 TI - Simuliidae and the transmission and control of human Onchocerciasis in Latin America. AB - Factors that affect the propensity of a simuliid species to act as a host to Onchocerca volvulus and to naturally transmit this filarial worm in nature are discussed. Presence or absence of a cibarial armature is believed to be a major factor that has been previously overlooked and this is considered in relation to the choice of control methods currently advocated for onchocerciasis. The current epidemiological studies, transmission dynamics and relevant control measures are discussed for each onchocerciasis focus in Latin America. PMID- 15806244 TI - [The organization of health services: the comparison as contribution]. AB - This article discusses about a recent procedure in health care studies, the comparison as a methodology of analysis. The different analytical currents refer to a particular method of understanding health-disease process. They are: functionalism, the historical-materialism and the new currents. Their phylosophical and sociological basis, concepts, analysis instruments and purposes are showed here by a review of the principal works from representative authors as Navarro, Terris, Roemer, Fry, Illich, Capra and others. The paper suggests that comparative analysis can take two directions: the first is a operational approach for analysing the concrete situations of health's service organization, the second, a more conceptual one, aimed at identifying critical questions and international tendencies in health's systems. The recent discussion search for the overcoming of these dichotomies toward the progress of the production of knowledge and its effects in health's services organization. PMID- 15806245 TI - [Knowledge, opinions and cigarette smoking among teachers]. AB - Due to the importance that educational activities have in preventing the habit of smoking in boys and young men, a research was carried out in order to know the rates of predominance, knowledge and opinions teachers have on smoking. For this purpose, an enquiry was applied to 746 teachers from Osorno, Chile. The results showed a rate of 42.6% of smokers; considering specialization, the major predominance is found in woman teachers of Biology and Physical Education. Comparisons with predominance in other countries are established. It was determined that the level of knowledge is independent from the habit of smoking; on the contrary, opinions about different aspects related to smoking, are closely connected to the habit. We conclude that it is necessary to carry on a systematic training with in service teachers and specially in Teacher's Training Colleges. PMID- 15806246 TI - [The low quality of home-made oral rehydration solution in Salvador, Bahia (Brazil)]. AB - In 1988 a nation-wide campaign aiming to inform the Brazilian population about preparation and use of simple sugar/salt oral rehydration solution (ORS) was carried out. The campaign was massively shown by the media. This preliminary study assessed the quality of ORS prepared by 23 mothers of in-patient children from a pediatric hospital in Salvador, State of Bahia. Fourteen (60.9%) among the 23 mothers prepared solutions containing Na concentrations ranging from 30 to 80 mmol/L which is recommended by the World Health Organization. Eleven (47.8%) solutions contained glucose within the recommended range of 30 to 112 mmoI/L Only 6 (26%) out of the 23 mothers prepared ORS with simultaneously adequate Na and glucose concentrations. However, just 3 (13%) out of these 6 ORS also presented balanced electrolyte concentrations. Potential iatrogenicity due to high concentrations of Na and glucose was found in 30.4% and 43.5% of the solutions, respectively. These data raise serious concerns about the quality of home-made sugar/salt ORS, and therefore about its use as a safe agent in a campaign of diarrhoeal diseases control. PMID- 15806247 TI - [Medical-sanitary assistance: notes for an evaluation]. AB - This paper presents and discusses the role of the research "Medical and Sanitary Care" (Assistencia medico-sanitaria-AMS) developed by the IBGE (Brazilian Institute of Geography and Statistics) in the context of the Brazilian Health System Reform process. It points out the importance of stablishing a greater interaction between the IBGE and the user of this institution 's information system in such a way that users' needs can be more clearly expressed in order to achieve a more productive use of the available information. Finally, the Author suggests that there should be some improvements in the diffusion of these researches and in the access to IBGE 's data bases, for these issues constitute two major barriers for the health planning process. PMID- 15806249 TI - [Dengue-clinical classification]. PMID- 15806248 TI - [Use of the Information, Memory and Concentration (IMC) Test in the epidemiological study of senile dementia in Fortaleza,Ceara (Brazil)]. AB - The objective of the present paper was to verify the occurrence of senile dementia in the city of Fortaleza, Brazil. A screening test, the Information, Memory and Concentration Test (IMC), was applied to 865 persons (65 years old and older). The Test includes questions on personal identification, recent and remote memory and concentration. Dementia was determined as a function of the total score obtained by the person in the test previously examined as far as sensitivity and specificity are concerned. The results were analysed according to age, sex and social strata. It was demonstrated a prevalence of dementia of 8.4% in the sample studied with no significant difference in the proportions of cases among people 75 years old or older (9.3%) as compared to the group younger, 65 to 74 years old, (7.5%). The percentage of dementia was similar between men (8.7%) and women (8.3%). It was verified a progressive increase in the frequency of dementia ranging from 5.2% for the higher social classes (A/B) to 6.9% and 10.3% for medium (C) and lower (D/E) ones respectively, with a significant difference between the social classes A/B and D/E. These results, however, should be seen with caution because of the percentage of refuse especially in the higher, A/B (19.3%) and lower D/E (9.3%) social classes. PMID- 15806250 TI - [Managerial modernity: proposal of model for organization structure]. PMID- 15806251 TI - [Episodic acute schistosomiasis in Ilha de Itamaraca, state of Pernambuco (Brazil)]. PMID- 15806252 TI - [AIDS: an hypothesis on its psychological disturbances]. PMID- 15806254 TI - [Not Available]. PMID- 15806253 TI - [The illness narratives: suffering, healing and the human condition]. PMID- 15806261 TI - [Credentialing or proficiency?]. PMID- 15806262 TI - [How many procedures are necessary to achieve competency in colonoscopy?]. AB - BACKGROUND: Competency for colonoscopy implies technical and cognitive skills. The American Society for Gastrointestinal Endoscopy has suggested 100 supervised procedures might be necessary. There are no specific recommendations in Brazil. AIM: To evaluate technical progress of trainees during a regular colonoscopy training program. MATERIALS AND METHOD: Two gastrointestinal fellows at Federal University of Sao Paulo, SP, Brazil, were prospectively evaluated during first year training. The frequency and time of reaching the cecum, total procedure duration, ability to identify lesions and patient discomfort were recorded. RESULTS: Two hundred and seventy one colonoscopies were preformed by both fellows (fellow A: 186 and B: 85). Twenty-seven cases were excluded of obstructive lesions or previous surgery, leading 171 and 72 exams, respectively. The mean success rate of reaching the cecum was 82.5% and 56.9%, respectively. For the first 72 exams it was 72.2% and 56.9%. Fellow A reached the cecum in 76% of first 100 colonoscopies, improving to 91.5% after the 101 st. Mean time for trainees to complete the procedure was 17.7 and 23.5 minutes to fellows A and B. Trainee A took 19.8 and 14.7 minutes before and after the 100th colonoscopy. DISCUSSION: Success rate and time taken to reach the cecum progressively improved over the number of procedures with statistical significance. However, because of individual differences, 100 colonoscopies may be insufficient to acquisition of technical skills. CONCLUSION: Although analyzing the learning curve of two fellows only, we could notice a statistically significant improve in reaching the cecum with experience over time. However, depending on individual skills more than 100 procedures may be necessary during training. PMID- 15806263 TI - Capella's gastroplasty: metabolites and acute phase proteins changes in midline and bilateral arciform approaches. AB - BACKGROUND: Obesity has adverse health effects. Dietary reeducation does not seem to offer sustained weight loss. For appropriately selected patients, surgery may be beneficial. AIM: To evaluate early postoperative metabolic response to surgery in patients submitted to Capella's gastroplasty using two different surgical approaches to the abdominal cavity. PATIENTS/METHOD: Twenty patients (9 males and 11 females, aged 21 to 53 years) were randomized prior to submission to either one of the surgical access incisions (bilateral arciform or supra-umbilical midline incisions). Blood samples were collected at the beginning and end of the operation, 12 (T-12 h) and 24 hours (T-24 h) postoperatively. Dieresis and synthesis time, blood loss, planimetry of operative field, operative time, hospital stay, hemoglobin, hematocrit, lymphocytes, potassium, albumin, erythrocyte sedimentation rate, C-reactive protein, glucose, pyruvate, lactate and ketone bodies were analyzed. RESULTS: Dieresis time was significantly decreased when median approach was used. Total operating time, hospital stay, hematocrit, hemoglobin, lymphocyte count, potassium and albumin concentrations were similar in both groups. C-reactive protein (T-12 h), glucose and pyruvate concentrations (T-24 h) increased significantly after completion of surgical procedure. Ketone bodies concentrations were significantly decreased 24 hour following completion of surgical procedure. CONCLUSION: Capella's gastroplasty induces metabolic and inflammatory changes in blood parameters. There is no evidence of technical superiority of arciform over midline incisions in this study. PMID- 15806264 TI - [Nutritional assessment in patients with cirrhosis: the use of indirect calorimetry]. AB - BACKGROUND: Malnutrition is frequent in cirrhotic patients, and its assessment is difficult. Functional assessment through a dynamometer is a simple method and could minimize these drawbacks. Harris-Benedict prediction formulae estimates the resting energy expenditure but has not been validated for this population. One alternative is the use of indirect calorimetry. AIM: To assess nutritional status in cirrhotic patients and estimates the resting energy expenditure through indirect calorimetry and compares it to Harris-Benedict. PATIENTS AND METHODS: Thirty four adult hepatitis C cirrhotic outpatients were studied, classified by Child-Pugh and model of end-stage liver disease score. The resting energy expenditure was predicted through Harris-Benedict and measured by indirect calorimetry. Nutritional assessment was done through anthropometry, subjective global assessment, hand-grip strength and a 3-day recall. RESULTS: Fifteen (44.2%) were Child-Pug A, 12 (35.3%) B and 7 (20.6%) C, and 33 (97.1%) had model of end-stage liver disease scores less than 20. The resting energy expenditure predicted was higher than the measured (Harris-Benedict 1404.5 +/- 150.3 kcal; indirect calorimetry 1059.9 +/- 309.6 kcal). The prevalence of malnutrition varied between methods (body mass index, muscle arm circumference, subjective global assessment, triceps skinfold thickness and hand-grip strength: 0; 5.9; 17.6; 35.3 and 79.4%, accordingly). Calories and proteins intake were 80% and 85% of recommended amounts and there was inadequate intake of calcium, magnesium, iron and zinc. CONCLUSION: Malnutrition was frequent and hand-grip strength seemed to be the most sensitive method for its diagnosis. Calories and protein intakes were inadequate. Considering that the predicted resting energy expenditure was higher than the measured one and the need to offer higher caloric intake, the use of the predicting equation may replace indirect calorimetry. PMID- 15806265 TI - [Immunohistochemical detection of p21ras, c-myc and p53 oncoproteins in hepatocellular carcinoma and in non-neoplastic liver tissue]. AB - BACKGROUND: Genetic and epigenetic alterations have been described in animal hepatocarcinogenesis models but need to be studied in human being. AIMS: To assess the immunoreactivity of p21ras, c-myc and p53 oncoproteins in hepatocellular carcinoma and non neoplastic tissue. Association of the immunoreactivity of these markers with histological grades and patterns, hepatitis B and C were additionally studied. METHODS: Detection of oncoproteins p21ras, c-myc and p53 was performed immunohistochemically in hepatocellular carcinoma (47 cases) and surrounding non neoplastic liver tissue (40 cases). RESULTS: Oncoproteins p21ras, c-myc and p53 were detected in 44,7%, 53,2% and 36,2% of the hepatocellular carcinoma cases, respectively. The p21ras and c-myc immunoreactivity has shown a significant association. However there was no association of p21ras, c-myc and p53 detection with hepatitis B and C virus infections, histological grades and patterns. The same significant association between p21ras and c-myc was observed in non-neoplastic tissue with cirrhosis when compared with tissue without it. The p53 immunoreactivity was negative in all non-neoplastic liver tissue samples. CONCLUSIONS: The immunoreactivity detection of p21ras, c-myc and p53 corroborates previous evidence of their detection in hepatocellular carcinoma that suggest the participation of these proteins in human hepatocarcinogenesis. The significant association between p21ras and c-myc oncoproteins in hepatocellular carcinoma and in cirrhosis can point to an interaction between them mainly, in hepatocarcinogenesis that occurs through cirrhosis. PMID- 15806266 TI - Liver resection: 10-year experience from a single institution. AB - BACKGROUND: Liver resection constitutes the main treatment of most liver primary neoplasms and selected cases of metastatic tumors. However, this procedure is associated with significant morbidity and mortality rates. AIM: To analyze our experience with liver resections over a period of 10 years to determine the morbidity, mortality and risk factors of hepatectomy. PATIENTS AND METHODS: Retrospective review of medical records of patients who underwent liver resection from January 1994 to March 2003. RESULTS: Eighty-three (41 women and 42 men) patients underwent liver resection during the study period, with a mean age of 52.7 years (range 13-82 years). Metastatic colorectal carcinoma and hepatocellular carcinoma were the main indications for hepatic resection, with 36 and 19 patients, respectively. Extended and major resections were performed in 20.4% and 40.9% of the patients, respectively. Blood transfusion was needed in 38.5% of the operations. Overall morbidity was 44.5%. Life-threatening complications occurred in 22.8% of cases and the most common were pneumonia, hepatic failure, intraabdominal collection and intraabdominal bleeding. Among minor complications (30%), the most common were biliary leakage and pleural effusion. Size of the tumor and blood transfusion were associated with major complications (P = 0.0185 and P = 0.0141, respectively). Operative mortality was 8.4% and risk factors related to mortality were increased age and use of vascular exclusion (P = 0.0395 and P = 0.0404, respectively). Median hospital stay was 6.7 days. CONCLUSION: Liver resections can be performed with low mortality and acceptable morbidity rates. Blood transfusion may be reduced by employing meticulous technique and, whenever indicated, vascular exclusion. PMID- 15806267 TI - [Clinical presentation of children with celiac disease attended at a Brazilian specialized university service, over two periods of time]. AB - BACKGROUND: Several studies have demonstrated changes in the forms of clinical presentation of celiac disease in children and adults, with an increased occurrence of atypical forms. AIM: To determine this fact in Clinical Hospital, Ribeirao Preto, SP, Brazil. METHODS: Celiac patients were studied over two different periods of time, from January 1978 to December 1987 (group 1 = G1) and from January 1988 to December 1997 (group 2 = G2). Time of disease, age at diagnosis and nutritional status and clinical forms were investigated. The typical form was considered to be present if the patient had at least two of the following signs/symptoms: diarrhea, abdominal distention, and weight loss, associated or not with others. RESULTS: Clinical forms (%) - G1: typical 57.14 and atypical 42.85. G2: typical 55.18 and atypical 44.82. Median age at diagnosis: 23 months (G1) and 21 months (G2). Interval between the onset of symptoms and the diagnosis: 13 months (G1) and 11 months (G2). Gomez nutritional status G1: Eutrophy = 23.8, DI = 47.61, DII = 23.8 and DIII = 4.76. G2: Eutrophy = 20.68, DI = 48.27, DII = 27.58 and DIII = 3.44. Waterloo G1: Eutrophy = 23.8, Wasting = 14.28, Stunting = 28.57 and Chronic = 33.33. G2: Eutrophy = 20.68, Wasting = 13.79, Stunting = 34.48 and Chronic = 31.03. No statistically significant differences occurred between the groups for any of these parameters. CONCLUSIONS: No change in the forms of presentation or remaining clinical characteristics of celiac disease occurred in our service. This may be due to the population characteristics or to differences among health services, or to different periods of observation. PMID- 15806268 TI - Evidence of mother-child transmission of Helicobacter pylori infection. AB - BACKGROUND: Low socioeconomical status is a major risk factor for natural acquisition of Helicobacter pylori (H. pylori) infection in developing countries. Its transmission route is unknown but studies suggest person-to-person transmission. AIM: To evaluate seropositivity of anti-H. pylori antibodies in family members of infected symptomatic index patients as compared to family members of symptomatic uninfected index patients. PATIENTS AND METHODS: One hundred and twelve family members of 38 patients who underwent endoscopy to exclude peptic disease were studied. Patients were deemed H. pylori infected or not infected when rapid urease test and histology were both positive or both negative. The family members underwent ELISA serology using the Cobas Core II Kit (Roche) and were classified into three groups: I - 29 family members of 10 H. pylori (+) duodenal ulcer index patients; II - 57 family members of 17 H. pylori (+) index patients without duodenal ulcer; III - 26 family members of 11 H. pylori (-) index patients. RESULTS: Seropositivity of group I and II (infected patients) was higher than the control group, 83% vs 38%, specially in mothers, 81% vs 18%, and in siblings 76% vs 20%. Differences between fathers' seropositivity was not statistically significant in the three groups: 100% vs 86% vs 70%. Seropositivity of all family members (mother, father and siblings) between infected group (I vs II) was similar. CONCLUSION: Prevalence of H. pylori infection was higher in family members of infected patients, but was similar among family members of infected patients with and without duodenal ulcer. H. pylori infection is more frequent in mothers and siblings of infected index children. A common source of infection cannot be excluded, but facts suggest that person-to-person transmission occurs, specially from mother to child. PMID- 15806269 TI - [Evaluation of postoperative adhesions in rats submitted to peritoneostomy with polypropylene mesh associated to nitrofurazone]. AB - BACKGROUND: Peritoneal injuries, common in per and postoperative, lead to the formation of adhesions. The use of polypropylene mesh increases the postoperative adhesions incidence. Based on the fact of nitrofurazone accelerates the healing process, a possible action on peritoneal adhesions was cogitated. AIM: To evaluate postoperative adhesions in rats submitted to peritoneostomy with polypropylene mesh fixation associated to nitrofurazone. METHODS: Thirty-three Wistar rats were separated into three groups of eleven animals each one. In group I was performed the laparotomy with exposition of the abdominal cavity followed by the laparotomy suture; in group II was performed the resection of a abdominal wall fragment followed by polypropylene mesh fixation on the gap and in group III was performed the same done in group II, but the mesh was previously impregnated with nitrofurazone. After 45 days the animals were sacrificed and necropsied. RESULTS: Seven animals of group I had no adhesions (degree 0), two had degree 1 adhesions and two had degree 2. In group II, one animal was classificated as degree 3 and ten as degree 4. In group III, one had degree 0, eight had degree 3 and one had degree 4 (one animal died before 45 days). All adhesions found involved the abdominal wall; ten animals had adhesions involving only omentum (group I = 4; group II = 1 and group III = 5); eleven animals had adhesions involving omentum, small bowel and other abdominal organs (group II = 10; group III = 1); three animals had adhesions involving only small bowel, all belonged to group III. Fourteen animals had adhesions involving small bowel, of these ten belonged to group II (nine with more than one fixation point) and four belonged to group III (three with just one fixation point). CONCLUSION: The association of nitrofurazone with polypropylene mesh did not reduce significantly the postoperative adhesions incidence in rats, but reduced the intensity and severity of these adhesions. PMID- 15806270 TI - [Hepatopulmonary syndrome: pathogenesis, diagnosis and treatment]. AB - BACKGROUND: The hepatopulmonary syndrome is characterized by a clinical triad consisting of: 1) hepatic disease and/or portal hypertension, 2) intrapulmonary vascular dilatations, and 3) abnormal arterial oxygenations (partial oxygen pressure <70 mm Hg or an alveolar-arterial oxygen gradient >20 mm Hg). Its frequency varies according to the diagnostic methods used. OBJECTIVES: To review the literature about the pathogeny, diagnosis and treatment of the hepatopulmonary syndrome. METHODS: A survey of the more relevant Brazilian and international publications on the hepatopulmonary syndrome in MEDLINE. RESULTS: The prevalence of the hepatopulmonary syndrome ranges from 4% to 17.5% depending on the diagnostic criteria used. The most commonly associated liver disease is cirrhosis. The alveolar-arterial oxygen gradient seems to be the best parameter for the assessment of the abnormalities of arterial oxygenation. For the detection of intrapulmonary vascular dilatations, contrast echocardiography is the method of choice because it is of easy execution and it can differentiate between the intrapulmonary and intracardiac communications. In the treatment of hepatopulmonary syndrome, it was possible to reproduce and confirm few successful reports using pharmacological therapy and/or intervention radiology. At present, liver transplantation is considered to be the main therapeutic option for these patients, with encouraging results. CONCLUSIONS: The hepatopulmonary syndrome is a frequent disease that requires blood gas measurements for diagnosis. Liver transplantation is the treatment of choice for patients with the syndrome. PMID- 15806271 TI - [Solid-pseudopapillary tumor of the pancreas: report of four cases]. AB - BACKGROUND: Solid-pseudopapillary tumor of the pancreas is a rare neoplasia of pancreas, with a rate varying from 0.17%-2.7% of non-endocrine tumors of the pancreas. Recently there has been a steady increase in the number of solid pseudopapillary tumors of the pancreas, with more than two thirds of the total cases described in the last 10 years. OBJECTIVE: To evaluate four patients with Frantz's tumor in a Brazilian institution. PATIENTS AND METHODS: Four patients with diagnosis of solid-pseudopapillary tumor were analysed in a retrospective analysis from December 2000 to February 2003, clinically and histopathologically. Therapeutical approach and prognosis were also studied. RESULTS: There were three females and one male with a median age of 27 years (range, 17-42). Dyspeptic symptoms and abdominal palpable mass represented the initial clinical findings. The main localization of the tumor was the pancreatic head (three of four cases). Enucleation was performed in one case, Whipple's surgery in two cases, and distal pancreatectomy in one case. Curative resection was possible in all cases confirmed by free margins. Two cases showed venous invasion histopathologically. Immunohistochemical analysis was done in three cases to confirm the diagnosis. In an average of 15-month follow-up, no recurrence has been observed. CONCLUSION: Our casuistic showed the preference of the tumor to the head of the pancreas, controversial to literature. However, other characteristics were similar to literature reports (clinical, histopathological, immunohistochemical and therapeutics). In addition, a longer postoperative follow-up will be necessary to affirm about prognosis. PMID- 15806272 TI - [Symptom's questionnaire for gastroesophageal reflux disease]. AB - BACKGROUND: Structured questionnaires are valuable instruments to measure the impact of specific diseases in patient's quality of life through a score and they are available such abroad as in Brazil. Nevertheless, questionnaires based on gastroesophageal reflux disease symptoms are not available in Portuguese. AIM: To develop and validate in Portuguese a specific questionnaire for gastroesophageal reflux disease symptoms. PATIENTS AND METHODS: Velanovich's original questionnaire was translated, one question about "regurgitation" symptom was included and the vocabulary was adjusted to be understood to the scholarity level of the analyzed population. The "face validity" to each question was evaluated by the members of a multidisciplinary panel and a symptom's questionnaire for gastroesophageal reflux disease was developed. The questionnaire was applied to patients with gastroesophageal reflux disease symptoms confirmed by prolonged pH esophageal monitoring. The reproducibility, the comprehension, the time spent to fill out the questionnaire and the correlation coefficient to Johnson-DeMeester's score were measured. RESULTS: The "face validity" was considered satisfactory by the panel and the questionnaire was applied to 124 patients, consecutively. The comprehension of the questionnaire and the time less than 5 minutes to fill out them were observed in all patients (100%). Reproducibility for 10 patients in two different occasions showed a high intra-class correlation coefficient of 0,833. The correlation to the Johnson-DeMeester's score was null. CONCLUSION: This study showed that the symptom's questionnaire for gastroesophageal reflux disease has "face validity", excellent reproducibility, easy comprehension and was quickly answered by patients. The correlation with Johnson-DeMeester's score was null. PMID- 15806273 TI - Hypercoagulable states: a review. AB - Vascular surgeons are often asked to evaluate patients with recurrent thromboses or thromboses in unusual locations either arterial or venous. There may be obvious inciting factors, but in many of these patients the possibility of a previously undiagnosed hypercoagulable state must be considered. Hypercoagulable states can be acquired (smoking, medication, change in physiologic status as occurs with pregnancy) or congenital. Congenital hypercoagulable disorders are more prevalent in families or patients with a history of thromboembolic events early in life or multiple miscarriages and include conditions such as protein C or S deficiencies, factor V Leiden mutation, and prothrombin gene variant. This article reviews the hypercoagulable states that are encountered in clinical practice as well as guidelines for testing and goals of therapy. PMID- 15806274 TI - Anastomotic strategies to improve hemodialysis access patency--a review. AB - The number of patients with end-stage renal disease (ESRD) who require maintenance hemodialysis has risen sharply in the past 2 decades. It is estimated that more than 60% of all patients with ESRD who require chronic hemodialysis are accessed through an arteriovenous fistula (AVF) or graft (AVG), and the incidence is increasing at a rate of 2% to 4% per year. The long-term patency rate of an upper extremity AVF or AVG for hemodialysis access remains suboptimal owing in part to progressive stenosis at the venous anastomosis. This article reviews the causative factors of dialysis access-related anastomotic stenosis, or intimal hyperplasia. This article also reviews the clinical experience of various anastomotic strategies to ameliorate the hemodynamic environment in an effort to improve the clinical outcome of hemodialysis access. These strategies include the use of (1) vein cuff at the expanded polytetrafluoroethylene (ePTFE)-venous anastomosis of AVG, (2) cuffed ePTFE dialysis AVG, and (3) anastomotic devices that create an interrupted anastomosis with staples or clips. PMID- 15806275 TI - Are we undertreating carotid stenoses diagnosed by ultrasound alone? AB - Clinical management of carotid disease is primarily based on results of national trials (ACAS and NASCET) that used the distal internal carotid artery diameter as a reference. However, commonly accepted ultrasound (US) criteria for carotid stenosis were derived from the correlation of velocity measurements with angiographic bulb diameter reductions (BDR). This study was undertaken to compare the degree of carotid stenosis determined by conventional velocity criteria to the degree of stenosis measured by B-mode (gray scale) diameter at both the carotid bulb and at the distal internal carotid artery, and, second, to evaluate US imaging to derive distal diameter reductions (DDR) noninvasively. During a 3 month period patients referred for carotid US were prospectively analyzed for standard velocity criteria and plaque morphology. Minimum carotid diameter was measured by longitudinal and transverse B-mode measurements and compared to carotid bulb diameter and internal carotid diameter distal to all disease. B-mode diameter reductions were compared to the degree of stenosis determined by velocity criteria and to patient symptoms and the decision for carotid endarterectomy. In total, 131 carotid arteries in 74 patients were evaluated. Based on the University of Washington velocity criteria, lesions were classified as grade I (n = 61, 46%), IIA (n = 58, 44%), IIB (n = 7, 5%), or III (n = 5, 4%). BDR measured by B-mode predicted the grade of disease based on velocity criteria (p < 0.001) with an overall accuracy of 95%. With use of the B-mode for DDR (NASCET style), 18 patients exceeded the 60% threshold for surgical intervention. Of these, only 3 patients were symptomatic and were operated on. An additional 3 operated-on patients had an asymptomatic grade III stenosis, our usual threshold for intervention. Twelve additional patients were appropriate for surgical intervention by B-mode but were not treated based on conventional velocity criteria alone. Bulb diameter reduction by B-mode imaging correlates strongly with diameter reduction determined by velocity criteria, and independently predicts the grade of carotid disease. With this in mind, the accuracy of B-mode imaging may be extended to the measurement of carotid stenosis based on DDR. By B mode criteria, many patients appropriate for intervention were not offered treatment based on conventional velocity criteria. Modern B-mode imaging provides a noninvasive method to obtain 'arteriographic equivalent'' measurements and should be added as a routine to carotid ultrasound interrogation. PMID- 15806276 TI - Arterial injuries from femoral artery cannulation with port access cardiac surgery. AB - Although minimally invasive (MI) cardiac surgery reduces blood loss, hospital stay, and recovery time, some MI approaches require femoral arterial cannulation, which introduces a heretofore unknown risk of femoral arterial injury. This study was performed to examine the risk of femoral arterial injury after Port Access MI cardiac surgery (PA-MICS) with femoral cannulation. Data were prospectively obtained on 739 consecutive patients who had PA-MICS with femoral cannulation between June 1996 and April 2000, identifying any patient with new (<30 days postoperative) arterial insufficiency from the cannulation site. Patient characteristics (gender, age, height, weight, body surface area, smoking, peripheral vascular disease, diabetes) and operative variables (cannula size, cross-clamp time) were examined with univariate and multivariate analysis to identify risk factors for arterial injury. Injuries were defined and classified by radiologic and intraoperative assessment, and follow-up was obtained by patient examination and from the medical records. Femoral arterial occlusion (FAC) occurred in 0.68% (5/739) of patients (4 women, 1 man; age range 26-74 years). The risk of femoral injury was higher in women: 1.31% vs 0.23% (p = 0.07). One patient had intraoperative limb ischemia from iliofemoral dissection and was treated by axillopopliteal bypass. Four patients presented postoperatively with claudication. Three of these had iliofemoral arterial occlusion or localized iliofemoral dissection and were treated with iliofemoral bypass, and 1 patient had localized femoral artery stenosis treated by angioplasty. With a mean follow-up of 17.8 months (range 13-26 months) limb salvage was achieved in all patients. Secondary or tertiary interventions were required in 40% (2/5), both in patients with iliofemoral occlusion, and 1 patient (20% of femoral injuries, 0.135% of overall series) has chronic graft occlusion and long-term claudication. The risk of arterial injury after femoral arterial cannulation and perfusion for Port Access surgery was low (0.68%). This risk is increased in women and is unpredictable. Initial vascular repair has a significant failure rate, and secondary interventions are often necessary. Although the femoral cannulation and perfusion technique is safe overall, the risk must be clearly recognized. PMID- 15806277 TI - Early transposition of the sartorius muscle for exposed patent infrainguinal bypass grafts. AB - The traditional approach for patent and exposed and infected infrainguinal bypass grafts in the groin has included wide operative debridement and secondary or delayed primary closure. However, this has been associated with significant risk of further contamination and length of stay. The authors reviewed their experience using the wide debridement, sartorius muscle flap transposition, and primary wound closure as an alternative. During the past 5 years, they have had 50 patients with major wound necrosis or infection in the groin or thigh with the graft or native artery being exposed after debridement. This group included 28 men; 74% of the patients had hypertension, 58% had diabetes, and 20% had renal failure. The grafts were split evenly between native vein and prosthetic material. After wide debridement, closure was performed by the vascular surgeon using the sartorius muscle flap. Postoperatively, there was an 8% major amputation rate and a 12% mortality rate in the first 30 days. One patient developed a pseudoaneurysm 5 weeks after placement of the flap. This patient underwent removal of the infected polytetrafluoroethylene graft with ligation of the common femoral artery. None of the procedures have resulted in further systemic or graft sepsis. None have resulted in arterial or graft blowout. Follow up was for an average of 18 months. Closure of groin and thigh wounds with exposed bypass graft or native artery can be safely performed with the sartorius muscle flap with excellent results. The length of stay of these patients compared to historical controls is acceptable. Furthermore, the chance of infection of the native artery or bypass may be reduced. Familiarity with this simple technique can be a valuable tool for the vascular surgeon. PMID- 15806278 TI - An integrative review of outcomes in patients with acute primary upper extremity deep venous thrombosis following no treatment or treatment with anticoagulation, thrombolysis, or surgical algorithms. AB - Primary upper extremity deep venous thrombosis (UEDVT) is a rare condition that typically affects young patients and can cause considerable long-term morbidity. Proposed treatments have included rest, heat, elevation of the affected limb, anticoagulation, thrombolysis, surgical decompression, percutaneous transluminal angioplasty (PTA), and stenting. However, the optimal management of primary UEDVT remains controversial. This study was an integrative review of the English language literature since 1965 on primary UEDVT, with comparison of long-term symptoms, rethrombosis, and pulmonary embolism in 4 treatment algorithms: rest, heat, and elevation alone; anticoagulation alone; surgical decompression without thrombolysis; and algorithms including thrombolysis. Forty-one studies describing 559 patients met the criteria for inclusion. Statistically significant differences were found among the 4 treatment algorithms in the incidence of residual symptoms (p < 0.000), the incidence of pulmonary embolism (p < 0.000), and the incidence of rethrombosis (p < 0.027). Residual symptoms and the severity of residual symptoms were greatest in the rest, heat, and elevation algorithm (74%), followed by the surgical (60%), anticoagulation (44%), and thrombolysis (22%) algorithms. Pulmonary embolism was also greatest in the rest, heat, and elevation algorithm (12%), followed by the anticoagulation (7%), thrombolysis (1%), and surgical algorithms (0%), while rethrombosis was greatest in the thrombolytic algorithm (7%) followed by the surgical (3%), anticoagulation (2%), and rest, heat, and elevation (0%) algorithms. These results support the current clinical practice of a staged, multidisciplinary approach to treatment of primary UEDVT that includes thrombolytic therapy and possible surgical decompression. Further studies are needed to evaluate the natural history of patients treated with thrombolysis alone, to assess the optimal timing of surgical decompression, and to determine the best use of PTA and stenting in the multidisciplinary approach. PMID- 15806279 TI - Decreased production of nitric oxide by peripheral blood mononuclear cells of patients with peripheral vascular disease. AB - Although prior studies have implicated nitric oxide (NO), a molecular messenger, in the development and progression of atherosclerosis, most of these studies have centered on atherosclerotic plaques. The current investigation determines whether a correlation exists between the presence of altered levels of NO production by peripheral blood mononuclear cells (PBMCs) and atherosclerotic disease. Venous blood was collected from 8 surgical patients having severe peripheral vascular disease and 8 healthy controls. PBMCs were separated by gradient centrifugation, diluted to 10(5) cells per mL, and cultured. Lipopolysaccharide (LPS), at doses of 10, 25, and 50 ng/mL, was used to stimulate NO production. Total nitric oxide assay was performed to determine the levels of NO produced by PBMCs at 24 and 48 hours. When stimulated by LPS there was an increase in NO production in the PBMCs cultured from control as well as patient samples, as compared to basal NO levels. However, the data demonstrate a significant decrease in the nitric oxide production in the patients with atherosclerosis as compared to that in the control group (p < 0.05). The differential production of nitric oxide by PBMCs of patients with atherosclerotic disease and healthy controls not only suggests that it has a role in the pathogenesis of this disease but also underlines its systemic nature. Blood cells circulating in the body with altered levels of NO production could have profound effects in the microvascular environment mediating molecular pathways and signaling cascades that activate and augment atherosclerosis. PMID- 15806280 TI - Endovascular repair of early rupture of Dacron aortic graft--two case reports. AB - Complications after open aortic surgery pose a challenge both to the vascular surgeon and the patient because of aging population, widespread use of cardiac revascularization, and improved survival after aortic surgery. The perioperative mortality rate for redo elective aortic surgery ranges from 5% to 29% and increases to 70-100% in emergency situation. Endovascular treatment of the postaortic open surgery (PAOS) patient has fewer complications and a lower mortality rate in comparison with redo open surgical repair. Two cases of ruptured abdominal aortic aneurysm (AAA) were managed with the conventional open surgical repair. Subsequently, spiral contrast computer tomography scans showed reperfusion of the AAA sac remnant mimicking a type III endoleak. These graft related complications presented as vascular emergencies, and in both cases endovascular aneurysm repair (EVAR) procedure was performed successfully by aortouniiliac (AUI) stent graft and femorofemoral crossover bypass. These 2 patients add further merit to the cases reported in the English literature. This highlights the crucial importance of endovascular grafts in the management of such complex vascular problems. PMID- 15806281 TI - Postcarotid endarterectomy pseudoaneurysm treated with combined stent graft and coil embolization--a case report. AB - Pseudoaneurysm formation is a rare complication following carotid endarterectomy (CEA); however, its occurrence is associated with significant risk of morbidity. The patient in this report presented 2 years following CEA with headache and lateral neck mass. The diagnosis of a 3.5 x 3.0 cm carotid artery bifurcation pseudoaneurysm was made by using magnetic resonance angiography (MRA). Endovascular exclusion of the aneurysm was accomplished with coil embolization of the external carotid artery followed by deployment of a 7 x 50 mm wall stent graft into the common carotid artery-internal carotid artery (CCA-ICA). The patient's symptoms improved and at 6-months postexclusion, duplex ultrasound demonstrated a significant reduction in pseudoaneurysm size. This case highlights the feasibility and safety of using endovascular techniques in the treatment of post-CEA pseudoaneurysm. PMID- 15806282 TI - Exteriorization and control of a retained embolization coil in the carotid artery -a case report. AB - Advances in endovascular technology have enabled the development of complex techniques for the treatment of vascular conditions. Not surprisingly, the modern vascular surgeon will likely encounter unusual complications and will need to formulate plans for their management. In the current case report, the vascular surgery service was consulted to assist in the management of a retained embolization coil in the carotid artery. Relevant aspects of detachable coils are discussed and the successful management of this potentially hazardous complication is described. PMID- 15806283 TI - Endovascular management of isolated axillary artery aneurysm--a case report. AB - The purpose of this paper is to report a case of axillary artery aneurysm, successfully treated by percutaneous transfemoral self-expandable polytetrafluoroethylene (PTFE) covered Wallgraft stent. A 20-year-old man presented with sudden-onset, severe aching pain of left upper limb that occurred while doing isometric exercise. He had feeble left brachial and radial pulses. Angiography revealed a 21 x 14 mm left axillary artery aneurysm with a 15 mm neck. There was an organized clot in the midpart of the left brachial artery. The aneurysm was successfully closed with a self-expandable 10 x 30 mm Wallgraft endoprosthesis PTFE graft stent, via the right femoral route. A check angiogram at 6 weeks of follow-up showed a patent stent with no endovascular leak. At 1 year of follow-up, the patient was asymptomatic with normal upper limbs pulsations. Axillary artery aneurysm may be treated with a stent graft, which is an effective, rapid, and definite treatment and is an acceptable alternative to standard open surgical repair. PMID- 15806284 TI - Ureteroarterial fistula with ruptured anastomotic pseudoaneurysm: successful management with vascular exclusion, extra-anatomic bypass and nephrectomy--a case report. AB - Ureteroarterial fistula is a rare but life-threatening condition most often arising as a consequence of combined vascular and urologic pathology. Only about 70 cases are reported in the English literature. Principles of repair include complete vascular isolation, extra-anatomic bypass, and urinary stream diversion away from major vascular conduits. The case presented herein is only the second reported instance of fistulization to an anastomotic pseudoaneurysm of an iliopopliteal bypass. PMID- 15806285 TI - Candida sepsis following infected iliocaval thrombosis: a case report. AB - The authors report the case of a patient who developed a thrombosis of the right iliac vein involving also the inferior vena cava (IVC), in association with Candida sepsis. Despite adequate and prolonged antimycotic treatment, the patient recovered from the fungal sepsis only following the surgical removal of the infected thrombus. PMID- 15806286 TI - Induction of Epstein-Barr virus lytic replication by recombinant adenoviruses expressing the zebra gene with EBV specific promoters. AB - The latent Epstein-Barr virus (EBV) is found in the cells of many tumors. For example, EBV is detectable in almost all cases, and in almost all tumor cells, of non-keratinizing nasopharyngeal carcinoma. Activating the latent virus, which will result in its lytic replication and the death of tumor cells, is a potential approach for the treatment of EBV-associated cancers. In this study, three recombinant adenoviruses were constructed to express the Zebra gene, an EBV gene responsible for switching from the latent state to lytic replication. EBV specific promoters were used in order to limit Zebra expression in EBV-positive cells, and reduce the potential side effects. The EBV promoters used were Cp, Zp and a dual promoter combining both promoters, CpZp. The Zebra protein was detected in HEK293 cells as well as the EBV-positive D98-HR1 cells infected with recombinant viruses. An EBV lytic replication early antigen, EA-D, was also detected in infected D98-HR1, implying the initiation of lytic replication. In the cell viability assay, Zebra-expressing adenoviruses had little effect on EBV negative HeLa cells, while significantly reducing the cell viability and proliferation of D98-HR1 cells. The results indicate that EBV virus promoters can be used in adenovirus vectors to express the Zebra gene and induce EBV lytic replication in D98-HR1 cells. PMID- 15806287 TI - nifH promoter activity is regulated by DNA supercoiling in Sinorhizobium meliloti. AB - In prokaryotes, DNA supercoiling regulates the expression of many genes; for example, the expression of Klebsiella pneumoniae nifLA operon depends on DNA negative supercoiling in anaerobically grown cells, which indicates that DNA supercoiling might play a role in gene regulation of the anaerobic response. Since the expression of the nifH promoter in Sinorhizobium meliloti is not repressed by oxygen, it is proposed that the status of DNA supercoiling may not affect the expression of the nifH promoter. We tested this hypothesis by analyzing nifH promoter activity in wild-type and gyr- Escherichia coli in the presence and absence of DNA gyrase inhibitors. Our results show that gene expression driven by the S. meliloti nifH promoter requires the presence of active DNA gyrase. Because DNA gyrase increases the number of negative superhelical turns in DNA in the presence of ATP, our data indicate that negative supercoiling is also important for nifH promoter activity. Our study also shows that the DNA supercoiling-dependent S. meliloti nifH promoter activity is related to the trans-acting factors NtrC and NifA that activate it. DNA supercoiling appeared to have a stronger effect on NtrC-activated nifH promoter activity than on NifA-activated promoter activity. Collectively, these results from the S. meliloti nifH promoter model system seem to indicate that, in addition to regulating gene expression during anaerobic signaling, DNA supercoiling may also provide a favorable topology for trans-acting factor binding and promoter activation regardless of oxygen status. PMID- 15806288 TI - Identification of peptides inhibiting adhesion of monocytes to the injured vascular endothelial cells through phage-displaying screening. AB - Using oxidized low-density lipoprotein (LDL)-injured vascular endothelial cells (ECs) as target cells, peptides specifically binding to the injured ECs were screened from a phage-displaying peptide library by using the whole-cell screening technique after three cycles of the adsorption-elution-amplification procedure. Positive phage clones were identified by ELISA, and the inserted amino acid sequences in the displaying peptides were deduced from confirmation with DNA sequencing. The adhesion rate of ECs to monocytes was evaluated by cell counting. The activity of endothelial nitric oxide synthase (eNOS), and the expression levels of caveolin-1 and intercellular adhesion molecule-1 (ICAM-1) were determined by Western blotting. Six positive clones specifically binding to injured ECV304 endothelial cells were selected from fourteen clones. Interestingly, four phages had peptides with tandem leucine, and two of these even shared an identical sequence. Functional analysis demonstrated that the YCPRYVRRKLENELLVL peptide shared by two clones inhibited the expression of ICAM 1, increased nitric oxide concentration in the culture media, and upregulated the expression of caveolin-1 and eNOS. As a result, the adhesion rate of monocytes to ECV304 cells was significantly reduced by 12.1%. These data suggest that the anti adhesion effect of these novel peptides is related to the regulation of the caveolin-1/nitric oxide signal transduction pathway, and could be of use in potential therapeutic agents against certain cardiovascular diseases initiated by vascular endothelial cell damage. PMID- 15806289 TI - Expression of monomeric insulin precursor in Pichia pastoris and its conversion into monomeric B27 Lys destripeptide insulin by tryptic hydrolysis. AB - Monomeric B27 Lys destripeptide insulin (B27 Lys DTrI) was designed and produced from its precursor expressed in Pichia pastoris through tryptic hydrolysis instead of the less efficient tryptic transpeptidation. The monomeric B27 Lys DTrI precursor (MIP) was purified from a cultured medium of P. pastoris by a combination of hydrophobic, size-exclusion, and ion-exchange chromatography. The purified MIP was converted, by tryptic hydrolysis, to B27 Lys DTrI, which was then purified by ion-exchange chromatography to homogeneity as assessed by native gel electrophoresis, HPLC, amino acid composition, and electrospray mass spectrometric analysis. B27 Lys DTrI exhibited superior monomeric properties in size-exclusion chromatography. The yield of MIP was 200 mg per liter of culture, and the overall yield of purified B27 Lys DTrI from the crude MIP was 70%. The in vivo biological activity of B27 Lys DTrI as determined by the mouse convulsion assay was 21 U/mg, identical to that obtained by semisynthesis. PMID- 15806290 TI - Identification of the alternative promoters of the KChIP4 subfamily. AB - The subfamily of voltage-dependent potassium (Kv) channel interacting protein 4 (KChIP4) is made up of the auxiliary interacting protein of voltage-dependent potassium channels. In this study, the structure of four splicing variants of the human KChIP4 gene was analyzed. Three of the four isoforms of the KChIP4 gene, KChIP4.1, KChIP4.2 and KChIP4.4, were amplified from mouse and human fetal brain tissues by reverse transcription-polymerase chain reaction and then identified. Based on the bioinformatics analysis of the genomic sequences of the gene, we cloned and characterized two promoter fragments from the KChIP4 gene. One was a 325 bp fragment upstream of the 5' end of the KChIP4.1 mRNA sequence and the other was an 818 bp fragment located immediately at the 5' end of the KChIP4.4 variant. Both of them can initiate the transcription of the reporter gene in HT1080 cells and Sprague-Dawley (SD) rat fetal brain neurons, and they contain CG islands, except typical TATA boxes and CAAT boxes. This shows that the KChIP4 gene expression is regulated by an alternative promoter. PMID- 15806291 TI - A viral protein suppresses siRNA-directed interference in tobacco mosaic virus infection. AB - Plant viruses encode suppressors of post-transcriptional gene silencing (PTGS), an adaptive defense response that limits virus replication and its spread in plants. The helper component proteinase (HC-Pro) of the potato virus A (PVA, genus Potyvirus) suppresses PTGS of silenced transgenes. Here, the effect of HC Pro on siRNA-directed interference in the tobacco mosaic virus (TMV) was examined by using a transient Agrobacterium tumefaciens-based delivery system in intact tissues. It was shown that the interference effect was completely blocked by co infiltration with HC-Pro plus siRNA constructs in both systemic and hypersensitive hosts. In the system host, all plants agro-infiltrated with HC-Pro plus siRNA constructs displayed the same symptoms as the negative control. Meanwhile, TMV RNA accumulation was found to be abundant in the upper leaves using reverse transcriptase-PCR (RT-PCR) and Northern blot assays. On the contrary, plants agro-infiltrated with the siRNA construct alone were free of symptoms. Therefore, our study suggests that the transient expression of HC-Pro inhibited the siRNA-directed host defenses against TMV infection. PMID- 15806292 TI - Gene transcription profile in mice vaccinated with ultraviolet-attenuated cercariae of Schistosoma japonicum reveals molecules contributing to elevated IFN gamma levels. AB - Vaccination with ultraviolet-attenuated cercariae of Schistosoma japonicum induced protective immunity against challenge infection in experimental animal models. Our preliminary study on the transcription levels of IFN-gamma and IL-4 in splenic CD4+ T cells revealed that attenuated cercariae elicited predominantly a Th1 response in mice at the early stage, whereas normal cercariae stimulated primarily Th2-dependent responses. Further analysis on the gene profile of the skin-draining lymph nodes demonstrated that the levels of IFN-gamma were significantly higher in vaccinated mice than those in infected mice at day 4, 7 and 14 post-vaccination or post-infection. However, for IL-12 and IL-4, the potent inducers of Th1 and Th2 responses, respectively, as well as IL-10, there were no differences over the course of the experiment between the infected and vaccinated mice. To explore the underlying factors that may potentially contribute to elevated IFN-gamma in vaccinated mice, the mRNA profiles of the skin-draining lymph nodes at day 4 post-exposure were compared using oligonucleotide microarrays. Within the 847 probe sets with increased signal values, we focused on chemokines, cytokines and relevant receptors, which were validated by semi-quantitative RT-PCR. A comprehensive understanding of the immune mechanisms of attenuated cercariae-induced protection may contribute to developing efficient vaccination strategies against S. japonicum, especially during the early stage of infection. PMID- 15806293 TI - On-column refolding of an insoluble His6-tagged recombinant EC-SOD overexpressed in Escherichia coli. AB - The EC-SOD cDNA was cloned by polymerase chain reaction (PCR) and inserted into the Escherichia coli expression plasmid pET-28a(+) and transformed into E. coli BL21(DE3). The corresponding protein that was overexpressed as a recombinant His6 tagged EC-SOD was present in the form of inactive inclusion bodies. This structure was first solubilized under denaturant conditions (8.0 M urea). Then, after a capture step using immobilized metal affinity chromatography (IMAC), a gradual refolding of the protein was performed on-column using a linear urea gradient from 8.0 M to 1.5 M in the presence of glutathione (GSH) and oxidized glutathione (GSSG). The mass ratio of GSH to GSSG was 4:1. The purified enzyme was active, showing that at least part of the protein was properly refolded. The protein was made concentrated by ultrafiltration, and then isolated using Sephacryl S-200 HR. There were two protein peaks in the A280 profile. Based on the results of electrophoresis, we concluded that the two fractions were formed by protein subunits of the same mass, and in the fraction where the molecular weight was higher, the dimer was formed through the disulfide bond between subunits. Activities were detected in the two fractions, but the activity of the dimer was much higher than that of the single monomer. The special activities of the two fractions were found to be 3475 U/mg protein and 510 U/mg protein, respectively. PMID- 15806294 TI - Cloning and characterization of beta-carotene ketolase gene promoter in Haematococcus pluvialis. AB - The unicellular green alga Haematococcus pluvialis accumulates a highly valuable ketocarotenoid, astaxanthin, under various environmental stresses. beta-carotene ketolase (BKT) plays a key role in astaxanthin biosynthesis in H. pluvialis. In this paper, an approximate 700 bp 5'-flanking region of the bkt gene containing a putative promoter was cloned through walking upstream. The results of the sequence analysis showed that this bkt 5'-flanking region might have cis-acting elements such as sterol regulatory element (SRE-1)-like motifs, the C repeat/dehydration responsive element (DRE) and al-3 proximal element (APE)-like motifs, except for typical TATA and CCAAT boxes. The results of the beta galactosidase assay and the transient expression of lacZ driven by a series of sequential deletions revealed that a minimal promoter-like region might exist from -630 to -408 bp, and the highest promoter activity was observed to span the positions from -630 to -308 bp. The results of the site-directed mutagenesis of a C-repeat/DRE and two APE-like motifs in a promoter-like region (-630 to -308 bp) suggested that two APE-like motifs might be essential for transcriptional control of the bkt gene. PMID- 15806295 TI - Postnatal developmental expression of calbindin, calretinin and parvalbumin in mouse main olfactory bulb. AB - The distribution of calbindin, calretinin and parvalbumin during the development of the mouse main olfactory bulb (MOB) was studied using immunohistochemistry techniques. The results are as follows: (1) calbindin-immunoreactive profiles were mainly located in the glomerular layer, and few large calbindin immunoreactive cells were found in the subependymal layer of postnatal day 10 (P10) to postnatal day 40 (P40) mice; (2) no calbindin was detected in the mitral cell layer at any stage; (3) calretinin-immunoreactive profiles were present in all layers of the main olfactory bulb at all stages, especially in the olfactory nerve layer, glomerular layer and granule cell layer; (4) parvalbumin immunoreactive profiles were mainly located in the external plexiform layer (except for P10 mice); (5) weakly stained parvalbumin-immunoreactive profiles were present in the glomerular layer at all stages; and (6) no parvalbumin was detected in the mitral cell layer at any stage. PMID- 15806296 TI - Laser-assisted microdissection, techniques and applications in pathology (review). AB - Innovations in molecular medicine provided sophisticated tools for analysis of the pathogenesis of diseases. Much emphasis is put on examination of alterations in affected organ systems. However, since in vivo tissues are inherently complex mixtures of different cell types, specific molecular data on individual cell populations are difficult to obtain. The advent of laser assisted microdissection (LAM) now allows efficient isolation of pure cell populations and even of single cells from mixed tissues. The most promising aspect of LAM is its combination with different molecular downstream analyses of microdissected cells at the levels of the genome, the transcriptome and the proteome. This review focuses on the two basic techniques of laser-assisted microdissection and on its applications in molecular pathology. PMID- 15806297 TI - Inactivation of Ets 1 transcription factor by a specific decoy strategy reduces rat C6 glioma cell proliferation and mmp-9 expression. AB - Malignant gliomas represent the most aggressive tumours of the central nervous system and are characterised by both extensive proliferation and invasive growth. Matrix degrading proteases called matrix metalloproteinases (MMPs), particularly MMP-9, play a crucial role in glioma infiltration. The activity of these enzymes is regulated at different levels. In this regard, the control of transcriptional activity by specific transcription factors is believed to be very important. In the present study, we examined whether rat C6 glioma cells express the Ets 1 transcription factor and whether inhibition of Ets 1 by a specific decoy strategy affects C6 glioma cell proliferation and mmp-9 expression. We found that C6 glioma cells express Ets 1 and can efficiently be transfected with an Ets 1 specific decoy oligodesoxynucleotide (ODN). This ODN significantly reduces cell proliferation and mmp-9 expression, the latter in a dose-dependent manner. We conclude that inhibition of transcription factors, which play a role for glioma development and progression such as Ets 1 by specific decoy approaches, might represent useful tools for experimental therapeutic strategies against malignant gliomas. PMID- 15806298 TI - Aspirin blocks binding of photosensitizer SnET2 into human serum albumin: implications for photodynamic therapy. AB - Tin etiopurpurin dichloride (SnET2) is one of the photosensitizers under investigation to be used in photodynamic therapy of prostate cancer. The drug is delivered intravenously, transported in vivo by liposomes and plasma proteins and localized within the prostate. SnET2 exists in two tautomeric forms (I - closed ring, II - open ring) with I converting spontaneously into the more energetically stable form II at physiological pH. Up to approximately 50% of the drug can be carried by serum albumin, although this association can increase photo-bleaching and diminish the drug efficiency. Molecular modeling and force field calculations indicate that Sudlow Site I in human serum albumin (HSA) is the most probable binding site for both forms of SnET2, with the porphyrin moiety nestling between domains IIA and IB, and the esterolytic side group oriented toward domain IIIA of HSA. Other drugs, including aspirin, bind to the same part of HSA. SnET2 does not bind to HSA when pre-incubated with aspirin, which confirms that its place of binding to this protein must be located near Lys199. This observation could be exploited to improve photo-efficiency of SnET2 by finding drugs that could compete with the photosensitizer for binding into Sudlow Site I of HSA. PMID- 15806299 TI - Structural analysis of epolactaene derivatives as DNA polymerase inhibitors and anti-inflammatory compounds. AB - Epolactaene (compound 1), a neuritogenic compound found in human neuroblastoma cells, was found to show anti-inflammatory activity in vivo in this study. DNA polymerases and DNA topoisomerase II (topo II) were some of the major molecular targets of compound 1. Since the agent seems to be a potential pharmaceutical medicine, we synthesized derivatives chemically and obtained seven compounds, 1 to 7 to screen clinically more efficient epolactaene derivatives. A comparison of its structural derivatives revealed that the long alkyl side chain seemed to have an important role in the inhibitory effect. Notably, C18-alkyl chain conjugated epolactaene (compound 5) was the strongest inhibitor of DNA polymerase alpha, beta, lambda (pol alpha, beta, lambda) and topo II, with IC50 values of 13, 135, 4.4 and 5 microM, respectively, and 500 microg of compound 5 caused a marked reduction in TPA (12-O-tetradecanoylphorbol-13-acetate)-induced inflammation (inhibitory effect, 65.0%). Compound 5 did not influence the activities of plant or prokaryotic DNA polymerases, or of other DNA metabolic enzymes such as telomerase, RNA polymerase and deoxyribonuclease I. Based on these results, the relationship among the three-dimensional structure of epolactaene derivatives and the inhibition of polymerases and topo II, and anti-inflammation is discussed. PMID- 15806300 TI - Expression of thioredoxin in patients with Graves' disease. AB - Thioredoxin (TRX), which is a stress-inducible protein with redox-active disulfide structures, has various biological activities by regulating DNA binding of transcription factors in cells. In Graves' disease that is among the common diseases of the thyroid, endogenous stresses that are induced by excess thyroid hormones or antibodies against thyroid stimulating hormone (TSH) receptors are responsible for the pathogenesis. The objective of this study was to examine the expression of TRX and to determine whether TRX is responsible for the pathogenesis of Graves' disease. The thyroid follicular cells were shown to express both TRX and vascular cell growth factors (VEGF) in all of the patients with Graves' disease by immunohistochemistry. In contrast, the expression of TRX or VEGF was not found in any of the normal thyroids. Serum levels of TRX were significantly elevated in patients with Graves' disease regardless of their thyroid function compared to those in healthy donors (122+/-16 versus 37+/-5 ng/ml, p<0.0001). Consecutive administration of iodine resulted in not only a reduction in serum levels of free triiodothyronine (T3) but also an increase in serum levels of TRX in the patients. These findings suggest that release of intracellular TRX from thyroid follicular cells in response to iodine resulted in suppression of T3 production. Taken together, TRX is highly produced under stress in Graves' disease and involved in regulating production of thyroid hormones. The investigation of biological behavior of this molecule may greatly help to understand pathogenesis of Graves' disease functions. PMID- 15806301 TI - Up-regulation of beta-catenin in external auditory canal cholesteatoma. AB - The external auditory canal cholesteatoma (EACC) is a rare disease with hyperproliferation and destructive growth in the adjacent structures. Down regulation of beta-catenin (key component of the zonula adherens) is a pivotal factor for loose tissue integrity and invasiveness. Transforming growth factor beta1 (TGF-beta1) was reported to decrease beta-catenin in mammary epithelium. We investigated the abrogation of TGF-beta1 and beta-catenin expression in EACC culture cells. Cultured EACC-specimens were incubated with 6 micromol TGF-beta1 antisense. After 48 h, expression of beta-catenin was determined by means of immunohistochemistry. The cells showed an increased mural reactivity to beta catenin, and intracellular reactivity was unchanged. The untreated cells showed a loss of beta-catenin expression at the membranes. The predominant membranous location after treatment with TGF-beta1 antisense suggests increased tendency of the cells for tissue formation and strong cell-cell adhesion rather than migratory and invasive character, and thus TGF-beta1 antisense application is a useful therapeutical strategy. PMID- 15806302 TI - Isolation of five types of flavonol from seabuckthorn (Hippophae rhamnoides) and induction of apoptosis by some of the flavonols in human promyelotic leukemia HL 60 cells. AB - Five types of flavonol were isolated from seabuckthorn (Hippophae rhamnoides) and identified by mass, 1H- and 13C-NMR. The proliferations of human promyelotic leukemia HL-60 cells were inhibited as the concentrations of these flavonols were increased. The order of the extent of growth inhibition by the flavonols at a concentration of 20 microM is as follows: pentamethylquercetin > syringetin > isorhamnetin > quercetin > kaempherol > myricetin. Apoptotic morphological changes of the nucleus, including chromatin condensation were induced in the HL 60 cells treated with quercetin, kaempherol and myricetin, respectively, but not in the cells treated with the other flavonols. The fragmentations of DNA by quercetin, kaempherol and myricetin, respectively, to oligonucleosomal-sized fragments, a characteristic of apoptosis, were observed to be dose-dependent in the HL-60 cells. These findings suggest that growth inhibition by quercetin, kaempherol and myricetin, respectively, results from the induction of apoptosis by these flavonols. The other flavonols (pentamethylquercetin, syringetin and isorhamnetin) having methoxy (-OCH3) group inhibited more strongly than the above 3 flavonols without induction of apoptosis in the HL-60 cells. These findings suggest that mechanisms of growth inhibition by pentamethylquercetin, syringetin and isorhamnetin are different from the apoptosis caused by quercetin, kaempherol and myricetin. PMID- 15806303 TI - Expression of hepatocyte growth factor and its receptor c-met, correlates with severity of pathological injury in experimental alcoholic liver disease. AB - Expression of hepatocyte growth factor (HGF) and its receptor, c-met is up regulated in various forms of liver injury. This study evaluated the relationship between HGF and c-met expression and pathological changes in experimental alcoholic liver disease. Rats (5 per group) were fed ethanol and a diet containing saturated fat corn oil or fish oil by intragastric infusion. Dextrose isocalorically replaced ethanol in controls. In a second set of experiments, Kupffer cells, endothelial cells and hepatocytes were isolated from rats in each group. Pathological evaluation and analysis of HGF and c-met expression were performed in liver and the different cell types. Increased expression of HGF and c-met expression was detected in the liver of rats showing necroinflammatory changes. The Kupffer and endothelial cells were primarily responsible for the increase in HGF, c-met expression was seen only in hepatocytes. Thus, up regulation of HGF and c-met occurred in the presence of the necrosis and inflammation suggesting that HGF may be acting to protect against liver injury or accelerate the regenerative process. PMID- 15806304 TI - Down-regulation of dopamine transporter and abnormal behavior in STAT6-deficient mice. AB - To determine the role of STAT6 transcription factors in brain function, we performed a battery of mouse behavioral analyses of STAT6-deficient mice and made comparisons with wild-type mice. STAT6-deficient mice were significantly more hyperactive in the final two blocks of an open field test, indicating abnormal habituation in completing the task. Two learning tasks, a water maze and a passive avoidance test, were mastered by STAT6-deficient mice as effectively as the wild-type. RT-PCR analysis suggested that the levels of dopamine transporter (DAT) mRNA may be lower in the midbrain of the mutant. In agreement with the finding, subsequent Western blotting and immunohistochemical analysis demonstrated lower levels of DAT protein in the mutant striatum. These results lead to the proposal that, in addition to being a pivotal transcription factor in the immune system, STAT6 may be crucially implicated in mouse behavior by modulating the expression of neuronal genes such as DAT. PMID- 15806305 TI - Estrogen specifically stimulates expression and production of osteoprotegerin from rheumatoid synovial fibroblasts. AB - We studied the effects of estrogen on human fibroblast-like synovial cells in rheumatoid arthritis (RA-FLS) focusing on receptor activator of NF-kappaB ligand (RANKL) and its decoy receptor osteoprotegerin (OPG), the osteoclast formation and function regulators that have a substantial role in bone erosion of RA. Estrogen influences osteoporosis and the onset of RA clinically. The cellular responses of RA-FLS to estrogen are initiated via two high-affinity estrogen receptors (ERs). Culture of RA-FLS in the presence of 10(-6) M 17beta-estradiol (E2) increased expression of estrogen receptor (ER)-alpha, but not ER-beta. OPG mRNA expression was significantly increased, whereas RANKL mRNA was unaffected. E2 treatment also significantly increased the amount of OPG released in the culture supernatant. The increase of OPG and ER-alpha was specifically antagonized by the pure estrogen antagonist ICI 182780. Tamoxifen, a selective ER moderator, did not increase OPG. The results indicate that estrogen stimulates secretion of OPG from RA-FLS by acting on ER-alpha, which likely prevents bone erosion in RA. PMID- 15806306 TI - IFN-gamma/JAK/STAT pathway-induced inhibition of DR4 and DR5 expression on endothelial cells is cancelled by cycloheximide-sensitive mechanism: novel finding of cycloheximide-regulating death receptor expression. AB - The pathway of interferon gamma (IFN-gamma-induced suppression in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated endothelial cell apoptosis was investigated. rTRAIL triggered apoptosis of human umbilical vein endothelial cells (HUVECs) in a type II cell death manner. IFN-gamma pretreatment significantly suppressed the expression of death receptor 4 (DR4) and DR5 on HUVECs, and inhibited apoptosis in response to TRAIL. IFN-gamma rapidly phosphorylated signal transducers and activators of transcription 1 (STAT1) and STAT6 but did not enhance phosphorylation of STAT3, Akt and extracellular signal regulated kinase (ERK) and nuclear translocation of NF-kappaB p65. Janus kinase (JAK)-induced phosphorylation of STAT1/6 appeared to be crucial since chemical inhibition of JAK abolished phosphorylation of STAT1/6, down-regulation of DR4/DR5 expression and IFN-gamma-induced inhibition of TRAIL-mediated apoptosis. IFN-gamma/JAK/STAT-induced suppression was regulated by cycloheximide (CHX) sensitive mechanism since the use of CHX mimicked the action of chemical inhibition of JAK in regard to DR4/DR5 expression as well as TRAIL-mediated endothelial cell apoptosis. We have not yet clarified precise mechanism, however, the present data provide a novel finding that IFN-gamma/JAK/STAT pathway elicits inhibition of TRAIL-mediated endothelial cell apoptosis through CHX-sensitive suppression of DR4/DR5. PMID- 15806307 TI - Effects of met-enkephalin on cell proliferation in different models of adrenocortical-cell growth. AB - Met-enkephalin (met-Enk) is an opioid peptide that acts via three main subtypes of receptors referred to as mu (mu)-, delta (delta)- and zeta (zeta)-receptor. While the first two receptor subtypes mediate the classic opioid effects of met Enk, zeta-receptors are reported to be involved in the non-opioid actions of the peptide, i.e. the inhibitory effect on the cell growth. Despite the fact that met Enk is known to regulate the function of the hypothalamic-pituitary-adrenal axis acting on both its central and peripheral branches, none is known on the effects of met-Enk on adrenal growth. Hence, we have investigated the effects of met-Enk and its receptor agonists and antagonists on cell proliferation in three different models of rat adrenal growth: i) immature adrenal cortex, ii) regenerating adrenal cortex and iii) primary cultures of adrenocortical cells. In in vivo experiments, rats were given subcutaneous injections of 1 nmol/100 g of the peptides 28, 16 and 4 h before the sacrifice, and proliferative activity was assessed by counting the number of metaphase-arrested cells (after vincristine administration). In in vitro studies, cultured adrenocortical cells were exposed for 48 h to the peptides at a concentration of 10(-6) M, and proliferative activity was measured by the EZ4U method. The blockade of mu- and delta-receptors raised proliferative activity in immature adrenals and decreased it in regenerating glands, and the effects were reversed by mu- and delta-receptor agonists. Naltrexone-induced blockade of all met-Enk receptor subtypes decreased proliferative activity in immature adrenal and raised it in regenerating glands. The exposure to either mu- or delta-receptor agonists and antagonists evoked doubtful or no effects on the proliferative activity of cultured adrenocortical cells. In contrast, met-Enk exerted a marked antiproliferogenic effect that was reversed by naltrexone. Taken together, these findings allow us to draw the following conclusions: i) mu- and delta-receptor activation inhibits the growth of immature adrenals, stimulates adrenal regeneration, and does not affect proliferation of cultured adrenocortical cells; ii) zeta-receptors mediate the growth inhibitory effect of met-Enk on both regenerating adrenals and cultured adrenocortical cells, but unexpectedly their activation stimulates the growth of immature gland; and iii) the effects of mu- and delta-receptor activation in in vivo experiments are probably mediated by extra-adrenal indirect mechanisms. PMID- 15806308 TI - Orexins modulate the growth of cultured rat adrenocortical cells, acting through type 1 and type 2 receptors coupled to the MAPK p42/p44- and p38-dependent cascades. AB - Orexin A and B are hypothalamic peptides that act through two subtypes of receptors named OX1-R and OX2-R. The OX1-R almost exclusively binds orexin-A, whereas OX2-R is non-selective for both orexins. We previously found that rat adrenocortical cells express both orexin-receptor subtypes, and orexin-A stimulates corticosterone secretion from dispersed adrenocortical cells acting via the OX1-R. Here, we examined the possibility that orexins, acting through both their receptor subtypes, modulate the growth of adrenocortical cells. Reverse transcription-polymerase chain reaction showed that rat adrenocortical cells cultured in vitro for four days expressed OX1-R and OX2-R mRNAs. Orexin-A increased the proliferation rate (PR) of cultured cells, while orexin-B lowered it. Using selective antibodies, we demonstrated that OX1-R immuno-blockade reversed the proliferogenic action of orexin-A, causing a sizeable decrease in PR. In contrast, OX2-R immuno-blockade magnified the proliferogenic effect of orexin-A and annulled the antiproliferogenic action of orexin-B. The proliferogenic effect of orexin-A in the presence of OX2-R immuno-blockade was abrogated by the MAPK p42/p44 inhibitor PD-98059, while the antiproliferogenic effect of orexin-A in the presence of OX1-R immuno-blockade was annulled by the MAPK p38 inhibitor SB-203580. Neither inhibitor altered per se the basal PR of cultured cells. Taken together, our present findings allow us to conclude that i) orexins modulate the growth of rat adrenocortical cells cultured in vitro, by exerting both proliferogenic and antiproliferogenic effects, which are mediated by OX1-Rs and OX2-Rs, respectively; and ii) OX1-R and OX2-R growth effects involve the activation of the MAPK p42/p44 and p38 signaling cascades, respectively. PMID- 15806309 TI - Overexpression of regucalcin suppresses apoptotic cell death in the cloned rat hepatoma H4-II-E cells induced by a naturally occurring isothiocyanate sulforaphane. AB - The effect of regucalcin, a regulatory protein in intracellular signaling system, on cell death and apoptosis was investigated. Sulforaphane, a naturally occurring isothiocyanate, is known to induce cell cycle arrest and apoptosis in cancer cells, although its effect has not been clarified in the cloned rat hepatoma H4 II-E cells. Hepatoma H4-II-E cells (wild-type) and stable regucalcin/pCXN2 transfected cells were cultured for 72 h in a medium containing 10% fetal bovine serum (FBS). Cells with subconfluency were changed to a medium containing either vehicle or sulforaphane (10(-7) or 10(-6) M) in the absence of FBS. After medium change, cells were cultured for 24, 48, or 72 h. The number of wild-type cells was significantly decreased in the presence of sulforaphane (10(-7) or 10(-6) M). Agarose gel electrophoresis showed the presence of low-molecular-weight deoxyribonucleic acid (DNA) fragments of adherent wild-type cells cultured with sulforaphane (10(-7) or 10(-6) M) for 24 h. Sulforaphane (10(-7) or 10(-6) M) induced cell number and DNA fragmentation was significantly suppressed in transfectants. The effect of sulforaphane (10(-6) M) in decreasing the number of wild-type cells was significantly prevented in the presence of caspase-3 inhibitor (10(-9) M), while the presence of Nomega-nitro-L-arginine methylester (NAME), an inhibitor of nitric oxide synthase, did not prevent sulforaphane induced death of wild-type cells. Sulforaphane (10(-6) M) did not have a significant effect on cell number of transfectants in the presence of caspase-3 inhibitor or NAME. This study demonstrates that sulforaphane induces cell death and apoptosis in the cloned rat hepatoma H4-II-E cells, and that overexpression of regucalcin suppresses sulforaphane-induced apoptotic cell death which is partly mediated through caspase-3.. PMID- 15806310 TI - Signal transduction in human endothelial cells induced by their interaction with group B Streptococci. AB - The molecular mechanisms underlying entry of group B Streptococci (GBS) into human endothelial cells are not yet fully understood. This study is centered on the triggering of signaling cascade in human umbilical vein endothelial cells (HUVEC) during their interaction with different GBS serotypes/strains (type III: 80340-vagina and 90356-CSF and type V: 88641-vagina and 90186-blood). We have shown that the analyzed microorganisms adhere to HUVEC, but only those of the strains 90356-CSF, 88641-vagina and 90186-blood presented intracellular viability. Activation of PKC directly increased F-actin content and organization into stress fibers, and increased intracellular viability of GBS-III microorganisms. PKA inhibitor seems to promote surveillance of GBS type V microorganisms within HUVEC. These studies indicate that different molecules present at the cell surface of the GBS might induce different responses to HUVEC, interfering with the recruitment of cortical actin filaments. PMID- 15806311 TI - Polymorphisms in the promoter regions of FAS and FASL genes as candidate genetic factors conferring susceptibility to endometriosis. AB - Although the pathophysiological mechanisms leading to endometriosis remain unknown, several hypothesis have been proposed, including a dysregulation of the normal apoptotic process which takes place in the endometrium. One of the apoptotic pathways playing a crucial role in the programmed cell death within the endometrium is the Fas-FasL system. In this study we have performed a case control analysis in order to evaluate three polymorphisms located within FAS ( 1377G>A and -670A>G) and FASL (-843C>T) genes, as susceptibility factors for endometriosis. We have analysed a series of women with endometriosis compared respectively to a group of women without symptoms of the disease, and to a group of confirmed unaffected women. The genotyping of the three variants was carried out by Fluorescence Resonance Energy Transfer (FRET) technology, and statistical analysis was performed using chi2 test with Yates correction. Our results show that the differences in the distribution of the polymorphic variants were not statistically significant when the group of patients was compared to the other groups. Thus, it seems to indicate that the variants here analysed are not involved in the pathogenesis of the disease in our population. However this does not let us to completely exclude such genes as potential candidates for the disease. A complete genetic analysis of the genes involved in the intricate regulatory system of the apoptosis may lead to the identification of susceptibility factors for the disease and a better understanding of its etiology. PMID- 15806312 TI - Na+,K+-ATPase activity is inhibited in cultured intestinal epithelial cells by endotoxin or nitric oxide. AB - Na+K+-ATPase is an important enzyme serving vital functions in various mammalian tissues, including the intestine. We have previously documented that endotoxin (LPS) and nitric oxide (NO) can induce enterocyte injury in vitro. To examine whether alterations Na+,K+-ATPase activity might be involved in LPS- or NO induced enterocyte dysfunction, we carried out four series of experiments. The first set of experiments documented that LPS decreases IEC-6 Na+,K+-ATPase activity at concentrations as low as 0.10 microg/ml. The second set of experiments tested whether exposure of IEC-6 cells to the exogenous NO donor, S Nitroso-N-acetylpenicillamine (SNAP), would decrease IEC-6 Na+,K+-ATPase activity. The results of these experiments documented that SNAP significantly decreased IEC-6 Na+,K+-ATPase activity in a dose-dependent fashion at a threshold inhibitory concentration of 0.1 mM, and there was an inverse correlation between Na+,K+-ATPase activity and NO concentrations in the medium. Since enterocytes contain iNOS, and LPS can increase iNOS activity, the third set of experiments examined the relationship between LPS-induced inhibition of Na+),K+-ATPase activity and NO production by the IEC-6 cells. These results showed that LPS increased IEC-6 NO production in both a dose- and time-dependent fashion and an inverse correlation existed between LPS-induced NO production and decreased Na+,K+-ATPase activity. Addition of the NOS inhibitor, L-NNA, prevented the LPS induced decrease in Na+,K+ATPase activity, suggesting that NO is involved in the decrease of Na+,K+-ATPase activity observed in the IEC-6 cells incubated with LPS. One mechanism by which the increased NO concentrations could have contributed to the decrease in Na+,K+ATPase activity, after the addition of LPS or SNAP, is via the production of peroxynitrite during the reaction of NO with superoxide. This notion was supported by studies showing that SNAP- and LPS induced decreases in IEC-6 Na+,K+-ATPase activity could be blocked by adding superoxide dismutase to the medium. The last set of experiments tested whether the inhibition of Na+,K+-ATPase activity with the specific Na+,K+-ATPase inhibitor ouabain would increase the permeability of an IEC-6 monolayer. IEC-6 monolayer permeability was increased by ouabain, but only at a high concentration. In conclusion, these studies indicate that LPS or the NO donor, SNAP, inhibit Na+,K+-ATPase activity and this inhibition is at least partly related to peroxynitrite production. These studies also suggest that LPS-induced NO production by the IEC-6 cells decreases IEC-6 Na+,K+-ATPase activity in an autocrine fashion. PMID- 15806313 TI - Comparative genomics on Wnt11 gene. AB - Non-canonical WNTs activate the planar cell polarity (PCP) pathway to induce cell motility and metastasis, while canonical WNTs activate the beta-catenin-TCF pathway to induce carcinogenesis. WNT11 gene at human chromosome 11q13.5 encodes non-canonical WNT11 protein, which is applicable for regenerative medicine of heart diseases. Here, we identified and characterized rat Wnt11 gene by using bioinformatics. Rat Wnt11 gene, consisting of five exons, was identified within AC120107.3 genome sequence. Rat Wnt11 (354 aa) was a secreted protein with 24 conserved Cys residues and five Asn-linked glycosylation sites. Rat Wnt11 showed 99.4%, 97.5%, 84.5% and 76.0% total-amino-acid identity with mouse Wnt11, human WNT11, chicken wnt11 and zebrafish wnt11, respectively. Comparative proteomics revealed that the number of Asn-linked glycosylation sites increased during molecular evolution of Wnt11 orthologs. Comparative genomics revealed that exon 1, but not 5'-flanking region, was well conserved between rat Wnt11 and human WNT11 genes. Although conserved transcription-factor-binding site was not identified within 5'-flanking region of rat Wnt11 and human WNT11 genes, Nkx2-5 binding site within exon 1 was evolutionarily conserved among mammalian Wnt11 orthologs. Because Nkx2-5 and Wnt11 are key regulators of heart development, Wnt11 was predicted as a target gene of Nkx2-5 transcription factor during cardiac myocyte differentiation. This is the first report on rat Wnt11 gene as well as on comparative genomics for Wnt11 orthologs. PMID- 15806314 TI - Comparative genomics on Norrie disease gene. AB - DAND1 (NBL1), DAND2 (CKTSF1B1 or GREM1 or GREMLIN), DAND3 (CKTSF1B2 or GREM2 or PRDC), DAND4 (CER1), DAND5 (CKTSF1B3 or GREM3 or DANTE), MUC2, MUC5AC, MUC5B, MUC6, MUC19, WISP1, WISP2, WISP3, VWF, NOV and Norrie disease (NDP or NORRIN) genes encode proteins with cysteine knot domain. Cysteine-knot superfamily proteins regulate ligand-receptor interactions for a variety of signaling pathways implicated in embryogenesis, homeostasis, and carcinogenesis. Although Ndp is unrelated to Wnt family members, Ndp is claimed to function as a ligand for Fzd4. Here, we identified and characterized rat Ndp, cow Ndp, chicken ndp and zebrafish ndp genes by using bioinformatics. Rat Ndp gene, consisting of three exons, was located within AC105563.4 genome sequence. Cow Ndp and chicken ndp complete CDS were derived from CB467544.1 EST and BX932859.2 cDNA, respectively. Zebrafish ndp gene was located within BX572627.5 genome sequence. Rat Ndp (131 aa) was a secreted protein with C-terminal cysteine knot-like (CTCK) domain. Rat Ndp showed 100, 96.9, 95.4, 87.8 and 66.4 total-amino-acid identity with mouse Ndp, cow Ndp, human NDP, chicken ndp and zebrafish ndp, respectively. Exon-intron structure of mammalian Ndp orthologs was well conserved. FOXA2, CUTL1 (CCAAT displacement protein), LMO2, CEBPA (C/EBPalpha)-binding sites and triple POU2F1 (OCT1)-binding sites were conserved among promoters of mammalian Ndp orthologs. PMID- 15806319 TI - Role of hypothalamic 5'-AMP-activated protein kinase in the regulation of food intake and energy homeostasis. AB - Although obesity is an epidemic threat to general health worldwide, an effective treatment has yet to be found. Insights into weight-regulatory pathways will accelerate the identification of new molecular targets for anti-obesity agents. 5'-AMP-activated protein kinase (AMPK) is an enzyme activated during low cellular energy charge. In peripheral tissues, the activation of AMPK influences various metabolic pathways, including glucose uptake, glycolysis, and fatty acid oxidation, all of which help to re-establish a normal cellular energy balance. AMPK is also present in the neurons of the hypothalamus, a critical center in the regulation of energy homeostasis. Recent studies from our group and others have shown that many factors (alpha-lipoic acid, leptin, insulin, ghrelin, glucose, 2 deoxyglucose, etc.) cause an alteration in hypothalamic AMPK activity that mediates effects on feeding behavior. Hypothalamic AMPK also appears to play a role in the central regulation of energy expenditure and peripheral glucose metabolism. These data indicate that hypothalamic AMPK is an important signaling molecule that integrates nutritional and hormonal signals and modulates feeding behavior and energy metabolism. PMID- 15806320 TI - Relationship between X-inactivation and clinical involvement in Fabry heterozygotes. Eleven novel mutations in the alpha-galactosidase A gene in the Czech and Slovak population. AB - We have identified 21 different alpha-galactosidase A gene (GLA) mutations in 22 unrelated Czech and Slovak families with Fabry disease. Eleven of these mutations were novel (point mutations D93N, A135V, D155H, G171R, Q280K, G360S, Q330X, splicing errors c.194ins14, c.801ins36 and deletions c.674_732del59, g.3405_6021del2617). Genotyping of family members for family-specific mutations revealed 55 heterozygotes that manifested clinical symptoms of different severity. To examine the contribution of X-inactivation skewing to disease manifestation in Fabry heterozygotes, we have adopted the Mainz severity scoring scheme and compared the score values with the X-inactivation status in 39 carriers in an age-dependent manner. The age-score trendline of Fabry females who had a predominantly inactivated X-chromosome bearing a wild-type GLA allele (10 of 38 females) was markedly steeper than in the rest of the cohort. One female carrier with an inactivated mutated allele had a low score value when compared to the other heterozygotes of the same age. These data suggest that X-inactivation is indeed a major factor determining the severity of clinical involvement in Fabry heterozygotes. There was a statistically significant difference between the severity score values of heterozygotes with random and non-random X-chromosome inactivation at the 5% level of significance. Further studies will show if the degree of the wildtype allele inactivation will be useful as a predictive marker of severity of phenotype in Fabry heterozygotes. Although the correlation between X-inactivation skewing and presentation of the disease in Fabry heterozygotes has previously been suggested in the literature, this report is among the first attempts to examine this relationship systematically. PMID- 15806321 TI - SUMO wrestling with type 1 diabetes. AB - Post-translational modification of proteins by phosphorylation, methylation, acetylation, or ubiquitylation represent central mechanisms through which various biological processes are regulated. Reversible covalent modification (i.e., sumoylation) of proteins by the small ubiquitin-like modifier (SUMO) has also emerged as an important mechanism contributing to the dynamic regulation of protein function. Sumoylation has been linked to the pathogenesis of a variety of disorders including Alzheimer's disease (AD), Huntington's disease (HD), and type 1 diabetes (T1D). Advances in our understanding of the role of sumoylation suggested a novel regulatory mechanism for the regulation of immune responsive gene expression. In this review, we first update recent advances in the field of sumoylation, then specifically evaluate its regulatory role in several key signaling pathways for immune response and discuss its possible implication in T1D pathogenesis. PMID- 15806322 TI - Incidence and causes for failure of treatment of women with proven osteoporosis. AB - The purpose of this study was to examine factors influencing women's compliance with medical therapy for osteoporosis in Israel. The study population consisted of women diagnosed with osteoporosis using a DEXA examination in the year 2000. Medication purchases for a period of 13 months after the exam were analyzed. Data were gathered from the computerized data base of Clalit Health Care Services and phone interviews. Treatment was divided into specific drugs and food additives (preparation of calcium plus vitamin D). Full, intermediate, low and no compliance were defined as >80, 50-80, 25-50 and <25% of treatment days, respectively. In the study, 857 women with a mean age of 73.6 years were enrolled; 581 (67.8%) were interviewed by telephone. Most of women were undereducated and unemployed. Twenty percent did not purchase any specific drug; 4.5% did not purchase any drug or food additive for osteoporosis; 60% were fully or intermittently compliant (by purchasing) with drug therapy. The most frequent single specific drugs purchased were bisphosphanates and SERM: 62.3 and 76.3% purchased bisphosphanates and/or food additives, respectively. Multi-variant analysis showed that predictors of compliance were healthy women, compliant with calcium plus vitamin D preparations, and/or spending more on drugs. Reasons for low or non-compliance were inconsistent recommendations by various physicians and the side effects of specific drugs. The relatively high compliance rate of osteoporosis treatment may be attributed to the increase in awareness of its benefits. The effect of physicians on compliance needs further investigation. PMID- 15806323 TI - Evaluation of pulmonary function and quality of life in women with osteoporosis. AB - Osteoporotic vertebral fractures generally result in an increased kyphotic angle, which in turn may lead to disturbances in pulmonary function. The objective of the present study was to evaluate pulmonary function and quality of life in a group of osteoporotic patients. Fifteen women with osteoporosis and thoracic vertebral fractures (group 1), 20 women with osteoporosis without vertebral fracture (group 2) and 20 control women (group 3) were submitted to spirometry using a Vitatrace-130 SL spirometer and to an SF-36 quality of life questionnaire. Women with osteoporosis and vertebral fractures showed an increased kyphotic angle (median=60 degrees) and decreased forced vital capacity (group 1 vs. group 2, P =0.020; group 1 vs. group 3, P =0.039) and forced expiratory volume in 1 s (group 1 vs. group 2, P =0.008; group 1 vs. group 3, P =0.014) when compared with women without vertebral fractures or osteoporosis. A negative correlation was observed between thoracic kyphosis and the predicted value of expiratory forced volume in 1 s ( r =-0.713, P =0.003). No differences in the quality of life were detected between the three groups studied. We conclude that women with thoracic vertebral fractures have an increased kyphotic angle and present a decrease in lung volume. PMID- 15806325 TI - Ethical considerations on consent procedures for emergency research in severe and moderate traumatic brain injury. AB - Therapeutic trials in TBI are subject to principles of Good Clinical Practice (GCP), to national legislation, and to international and European ethical concepts and regulations [e.g. 13]. The guiding principles underlying these investigations of treatment are respect for autonomy of research subjects, protection against discomfort, risk, harm and exploitation and the prospect of some benefit. Patients with significant TBI are mentally incapacitated, thus prohibiting obtaining consent directly from the subject. Various approaches to consent procedures are used as surrogate to subject consent: proxy consent, consent by an independent physician and waiver of consent. These approaches are reviewed. A questionnaire soliciting opinions was mailed to 148 EBIC (European Brain Injury Consortium) associated neuro-trauma centers in 19 European countries. 48% respondents believe that relatives were not able to make a balanced decision, 72% believed that consent procedures are a significant factor causing decrease in enrollment rate and 83% stated that consent procedures delay initiation of study treatment, resulting in possible harm if the agent has shown to be effective. 64% of the respondents considered TBI an emergency situation in which clinical research could be initiated under the emergency exception for consent. In new European legislation, emergency research under waiver of consent is not permitted. Nevertheless, we consider that randomising patients with TBI into carefully evaluated trial protocols without prior consent may be considered ethically justified. PMID- 15806324 TI - Interaction between proliferating cell nuclear antigen (PCNA) and a DnaJ induced by DNA damage. AB - Proliferating cell nuclear antigen (PCNA) is an essential protein for both DNA replication and DNA repair. In the present study using two-hybrid analysis with PCNA from rice, Oryza sativa L. cv. Nipponbare (OsPCNA), we found that OsPCNA interacted with rice DnaJ protein. We have identified DnaJ and designated it as OsDnaJ. OsDnaJ was able to bind to OsPCNA in vitro. Transcripts of OsDnaJ were found to be strongly expressed in the proliferating cells. mRNA of DnaJ was induced by UV and DNA-damaging agents such as H2O2. The expression patterns of OsPCNA were almost the same as OsDnaJ. The relationship between OsPCNA and OsDnaJ is discussed. PMID- 15806326 TI - Aneurysm recurrence after treatment of paraclinoid/ophthalmic segment aneurysms- a treatment-modality assessment. AB - OBJECT: The treatment of 21 paraclinoid/ophthalmic segment internal carotid artery aneurysms (PCOSAs) over a seven year period in a regional neurosurgical center was reviewed to assess the degree of obliteration and recurrence rate of these aneurysms after treatment by surgical and endovascular methods. METHOD AND RESULT: An assessment of the clinical notes, operation records and cerebral angiograms was made to evaluate the rate of residual and recurrent aneurysms after treatment and at follow-up. In the coiling group, the aneurysm recurrence rate was eight out of fifteen aneurysms (53%). Four recurrences were from previously totally occluded aneurysms. Out of the six surgical cases, five had follow-up angiography performed. All had stable occlusions of their aneurysms including one with subtotal occlusion. Two clipping procedures after previous coiling achieved total occlusion of aneurysm on follow-up angiography. CONCLUSION: Based on our case series we conclude that PCOSAs frequently recur after primary treatment. GDC coiling was associated with a higher rate of recurrent aneurysms when compared with surgical treatment. A review of the literature on the surgical and endovascular treatment of PCOSAs support this observation. PMID- 15806327 TI - Complete removal of ossification of the posterior longitudinal ligament in the mid-thoracic spine. PMID- 15806328 TI - Combined endoscopic surgery and radiosurgery as treatment modality for olfactory neuroblastoma (esthesioneuroblastoma). AB - OBJECT: Treatment of esthesioneuroblastoma (olfactory neuroblastoma) has been considerably improved by microsurgical techniques. Nevertheless, these rare tumours of the frontal skull base are still associated with high rates of tumour recurrence and mortality, thus remaining a challenge even for experienced surgeons. A novel therapeutic approach that combines endoscopic sinus surgery and Gamma Knife radiosurgery is presented here. Taking into account the rarity of the disease the present study comprises a relatively large series of patients treated in a similar manner. METHODS: 14 patients (8 males, 6 females) aged 27-75 years (median 38) were treated between May 1993 and December 2003. This series comprises 12 newly diagnosed esthesioneuroblastomas. Two more patients had already previously undergone surgery (24/39 months earlier). Paranasal and nasal endoscopic sinus surgery was performed. Marginal irradiation doses ranging from 15-34 Gy were given to the residual tumours by means of radiosurgery (Gamma Knife) involving 1-7 isocentres within 3 months after surgery. Median follow-up is 58 months (range 13-128). RESULTS: There was no mortality. In all patients tumour control was achieved within the treated area. 4 patients underwent a second radiosurgical procedure 6-79 months (median 34 months) after initial radiosurgery. One patients had to undergo an additional craniotomy because of extensive neoplastic infiltration, 1 developed postoperative liquorrhea, 1 case was complicated by bilateral frontal sinusitis. All patients complained of nasal discharge and crusts. Karnovsky Index--preoperatively ranging from 80%-100%- remained stable in 12 patients, an improvement was observed in 2 cases. CONCLUSIONS: Based on the favourable results recorded so far, the combination of endoscopic sinus surgery and radiosurgery can be considered a promising treatment option for esthesioneuroblastoma that merits further consideration. PMID- 15806329 TI - Fracture of anterior cervical plate implant--report of two cases. AB - Anterior cervical plate is a convenient and popular implant employed after corpectomy and bone grafting for traumatic or degenerative cervical spine disease. Although short-segment anterior cervical surgery may adequately be managed with anterior plate fixation alone, multilevel anterior cervical constructs exhibit a relatively high complication rate warranting a simultaneous posterior fusion. We report two patients undergoing two-level anterior corpectomy/fusion with a fixed anterior plate alone and exhibiting plate fracture in conjunction with pseudoarthrosis. The instrument failure was mainly attributed to pseudoarthrosis. However, improper contouring of the plate causing microstructural damage might create a weak point and contributed to this unusual hardware failure. PMID- 15806330 TI - Behcet's disease, associated with subarachnoidal heamorrhage due to intracranial aneurysm. AB - Behcet's disease is an unusual medical condition in central Europe and North America, however more common in Turkey and Japan. It was originally described in Turkey, characterized by recurrent oral ulcers, genital ulcers and also uveitis. A variety of vascular lesions such as venous occlusions, arterial aneurysms and varices account for the high rate of morbidity and mortality with this disease. Arterial aneurysms most commonly occur in the abdominal aorta, femoral arteries and in the pulmonary arteries. To our knowledge there have been seventeen documented reports of patient's with Behcet's disease combined with aneurysms of cerebral arteries. We describe a patient with Behcet's disease and subarachnoid haemorrhage due to a ruptured cerebral aneurysm. PMID- 15806331 TI - Presurgical octreotide treatment in acromegaly: no improvement of final growth hormone (GH) concentration and pituitary function. A long-term case-control study. AB - BACKGROUND: The effect of presurgical long-acting somatostatin analogue (SSA) treatment on operative outcome in acromegaly is as yet uncertain and long-term observations are lacking. We evaluated in an acromegaly case-control study the effect of octreotide pre-treatment on short- and long-term postoperative GH concentrations, pituitary function and glucose tolerance. METHODS: 48 patients with a pituitary macro-adenoma - micro- and giant adenomas excluded - were evaluated. 24 patients received presurgical octreotide treatment (secondary surgery, prospectively studied). Another 24 thoroughly matched patients had been operated on without prior octreotide therapy (primary surgery, retrospective evaluation). No patient had received any other treatment prior to operation/octreotide. Standardized testing was performed at diagnosis, following octreotide treatment, after surgery and then yearly for 10.3+/-0.9 yrs (mean+/ SE, primary surgery) and 4.1+/-0.6 yrs (secondary surgery). Immediate and 4-year postoperative results were compared. All work-up was strictly identical in both groups, except for imaging techniques. "Partial remission" was defined as mean GH profile (6-h/7-point) concentration <2.5 microg/L, and "complete remission" as GH nadir <1 microg/L during OGTT plus normal IGF-I concentration (when available). FINDINGS: The median profile GH (microg/L) values and the OGTT GH nadir values post-surgery (2.4/1.0 vs 1.8/0.7, primary and secondary surgery, resp.) as well as 4 yrs later (2.1/1.15 vs 2.3/0.8) were not significantly different between the groups. The 10-year results of the primary surgery group were not significantly different from its 4-year results. Subgroup analysis of pre-treated patients revealed no significant difference between those with and without tumour shrinkage, or between those with and without parasellar tumour extension. Postoperatively pituitary function was not significantly different between the groups. After 4-years the pituitary-adrenal axis was slightly more impaired in the secondary surgery group rather than following primary surgery, while the pituitary-gonadal axis was not different. CONCLUSION: Presurgical octreotide treatment has no significant short- or long-term beneficial effect on GH concentration or pituitary function. PMID- 15806332 TI - 3D ultrasound navigation in syrinx surgery - a feasibility study. AB - BACKGROUND: The prototype of a 3D ultrasound navigation system, with a trackable 4- to 8-MHz phased-array ultrasound probe was used in syrinx-surgery. The aim of this study was to evaluate the advantages offered by 3D ultrasound and navigation in syringomyelial surgery. METHODS AND MATERIALS: After laminectomy, with a free handed tilt of the ultrasound probe, the 3D volume of the target area is acquired within 15 seconds. The data are visualized after reconstruction in an axial, coronal, and sagittal view, offering the possibility of ultrasound-based guided surgery. RESULTS: Based on the intraoperative volume information, it was possible to navigate with the 3D ultrasound images in all cases. The orientation and image quality with respect to resolution, spatial information, and the identification of anatomical structures facilitated the surgery in all seven cases. The navigation tool, with a length of 12 cm and a tip diameter of 1 mm, was simple to place into the surgical site. The availability of an up-to-date 3D-image resulted in less interruption of the surgical procedure, with no need to repeatedly fill the cavity with sterile saline for new ultrasound acquisitions. New ultrasound images were only required if shift occurred. The coronal and "trajectory-plane" views, offer additional information about the syrinx cavity. The target borders are easier to determine and orientation in separated cavities was possible. Particularly in syringomyelial surgery it was helpful to determine the surface point of the syrinx to place the myelotomy or insert a catheter. CONCLUSION: 3D ultrasound offers the advantages of visualizing the third dimension of the target. For orientation and border determination navigation within the 3D ultrasound volume is very helpful and can take place with the ultrasound probe out of the way. Any disruption in the surgical procedure is minimized by not having to repeatedly fill the cavity with a sterile saline solution, there are fewer difficulties with image orientation because of new image adjustments. PMID- 15806333 TI - Varied patterns of postoperative course of disappearance of hemifacial spasm after microvascular decompression. AB - BACKGROUND: The precise course of resolution of postoperative residual HFS after a single MVD has not been well categorised quantitatively in the literature. Not all patients with HFS were cured immediately after a single MVD; some of them exhibited a gradual disappearance of the HFS over a period of time. The time course of the gradual resolution of HFS is categorised and the situation of re exploration in a few patients whose HFS persisted or recurred is determined. METHODS: The results of 547 MVDs in 545 Chinese patients were reviewed using the database from the author's personal records between January 1992 and December 2002. Different outcomes were observed and divided into four categories according to the variable convalescent period: type 1, spasm cured immediately; type 2, spasm persisted with milder severity and faded away gradually from 7 days to as long as 2 years; type 3, spasm ceased immediately but recurred after 3 days and ran the same course as in type 2; type 4, failed. FINDINGS: Four hundred and seventy-nine patients followed a type 1 course, which constituted an 87.9% immediate success rate. Forty-one patients (7.5%) followed a type 2 course, including three whose spasm persisted for more than 2 years. Twenty-three patients (4.2%), including one with venous compression, followed a type 3 course with their spasm ceased within three months. All of them had typical vascular loop compression. Another patient with a venous contact failed to respond positively to the first MVD underwent re-operation within 2 months. Re-operation was also performed in one patient four days after the first MVD due to persisting and even more severe spasm. These two patients were categorized type 4 as they failed the first MVD (0.4%). Late recurrence was noted in five patients from 1 to 2 years after the first MVD, only one of whom underwent re-exploration and was then cured again. CONCLUSIONS: Approximately 80% of the patients with HFS achieved immediate excellent results after a single MVD, the rest of the patients exhibited residual but usually milder spasms, which resolved gradually over a period of time. According to the patterns of the residual or persisting spasms, the situation and timing of re-exploration can be determined without difficulty. The surgeon should be concerned about missing the responsible vessels in patients with typical HFS who completely failed the first MVD. PMID- 15806334 TI - Genetic structure of Japanese scallop population in Hokkaido, analyzed by mitochondrial haplotype distribution. AB - To examine the genetic structure of Japanese scallop populations (Mizuhopecten yessoensis) in Hokkaido prefecture, Japan, and compare it with those in the Aomori prefecture, we applied a method for lineage analysis based on sequence variation in a mitochondrial DNA segment (NcR2). After showing that there was a low probability of doubly uniparental inheritance of mitochondrial DNA in the scallop, we sequenced the NcR2 regions of 914 individuals from 15 populations (13 in Hokkaido and 2 in Aomori). In total, 103 different haplotypes were detected. Results of homogeneity tests for pairwise populations and the fixation indices indicated that significant heterogeneity (P < 0.0005) and structuring (pairwise fixation index F(ST) = 0.1606-0.4444, P = 0.0000; fixation index among groups F(CT) = 0.1549, P = 0.0078) could be inferred between the Hokkaido and Aomori groups, but not among populations within the groups. Moreover, heterogeneity of the haplotype distribution between populations of the 1980s and 1990s or 2000s at the 4 culturing areas was not observed (P > 0.05), and the haplotype diversity between them was not significant (P = 0.05), suggesting that the culture operations had not imparted a significant effect on the genetic structure during these periods. PMID- 15806338 TI - Regional cerebral blood flow during auditory recall in 47 subjects exposed to assaultive and non-assaultive trauma and developing or not posttraumatic stress disorder. AB - OBJECTIVE: Psychological trauma leads to posttraumatic stress disorder (PTSD) in susceptible subjects. The aim of this study was to investigate the differences in regional cerebral blood flow (rCBF) between two groups of subjects exposed to different types of traumatic stressor either developing or not developing PTSD. METHODS: Twenty subjects developing (S) and 27 not developing (NS) PTSD after being exposed to either earlier person-under-the-train accident (NA) or being assaulted in the underground environment (A) were included in the study. 99mTc HMPAO SPECT was performed and the uptake in 29 regions of the brain (VOIs), bilaterally, was assessed. rCBF distribution was compared, using analysis of variance (ANOVA), between groups (S/NS) and type (A/NA) during a situation involving an auditory evoked re-experiencing of the traumatic event. Discriminant analysis was applied to test the concordance between clinical diagnosis and SPECT findings. RESULTS: In the general analyses significant differences were found between groups and types and there was a significant hemisphere x type interaction. S showed higher CBF than NS and so did A as compared to NA, particularly in the right hemisphere. Discriminant analysis correctly classified 66% of cases (p < 0001) in testing S/NS and 72% (p < 0001) in testing NA/A. CONCLUSIONS: Under recall of their traumatic experience we found higher relative CBF distribution values in S as compared to NS. CBF was higher in the right hemisphere and particularly in assaulted subjects. These findings underscore the role upon trauma recall of both the right hemisphere and the nature of the stressing event. PMID- 15806339 TI - Primary and secondary vasculitic neuropathy. AB - Necrotizing vasculitis occurs as a primary phenomenon in connective tissue disorders and cognate fields, including polyarteritis nodosa and the Churg and Strauss syndrome variant, rheumatoid arthritis, systemic lupus and Wegener's granulomatosis. In all these conditions focal and multifocal neuropathy occur as a consequence of destruction of the arterial wall and occlusion of the lumen of small epineurial arteries. Vasculitis may also complicate the course of other conditions ranging from infection with the HIV and with the B and C hepatitis viruses to diabetes and sarcoidosis. Pathologically polymorphonuclear cells are present in the infiltrates of the vessel wall in primary necrotizing vasculitis, while in secondary vasculitis the inflammatory infiltrate is mainly composed of mononuclear cells. In all instances symptomatic vasculitis requires corticosteroid to control the inflammatory process and prevent further ischemic nerve lesions. PMID- 15806340 TI - Occipital lobe epilepsy secondary to ulegyria. AB - OBJECTIVE: To analyse clinical and therapeutic aspects of epilepsy secondary to ulegyria in adults. PATIENTS: Out of 1,020 consecutive patients studied at a tertiary care epilepsy centre, eight cases of ulegyria were identified. All patients had comprehensive clinical evaluation, neuropsychological testing, interictal EEG, and brain magnetic resonance imaging (MRI). In addition, five patients had video-EEG monitoring. Ulegyria was confirmed by histological analysis in two patients who had successful epilepsy surgery. RESULTS: All patients had a history of perinatal asphyxia. In four of them there was psychomotor developmental delay. Mean age at onset of seizures was 5.8 years (range first week to 21 years). Brain MRI demonstrated predominant involvement of occipito-parietal cortical and subcortical areas. This posterior distribution of lesions was also supported by the presence of auras with occipital and parietal semiology in six patients, and signs of visuospatial dysfunction in five. Four patients had medically refractory epilepsy and two of them had significant improvement with surgical treatment. CONCLUSIONS: In this group of adult epileptic patients with ulegyria brain MRI, ictal semiology, and neurological examination are consistent with occipital lobe epilepsy. Most patients have severe epilepsy, but in some of them epilepsy can be controlled with antiepileptic drugs, while in others surgical treatment can be effective. Brain MRI criteria of ulegyria are well established, and in two cases it was possible to confirm their diagnosis with histological analysis. PMID- 15806341 TI - [Godfrey N. Hounsfield and his influence on radiology]. AB - On August 12, 2004 Sir Godfrey N. Hounsfield died at the age of 84 years. Computed tomography, which he developed, has been the most important invention in radiology since the discovery of X-rays; it revolutionized the medical diagnostic undisputedly. The essay gives an insight into the personality and the achievments of G. N. Hounsfield as well as the evolution of computed tomography of the classical sequential scanning via single to the current multislice spiral technique. The perspectives of computed tomography are shown. PMID- 15806342 TI - [Use of lubricants in urology. Indications and results]. AB - Lubricants are used for catheterization and/or endoscopic maneuvers. "The lubricant" should guarantee sufficient lubrification of the urinary tract, good visualization during endoscopy, and excellent local anesthesia. Additionally, asepsis or reliable control of the local bacterial flora of the urethra should be ensured. Modern lubricants fulfil these recommendations; in addition, they provide therapeutic opportunities for local therapy, for instance, of non gonococcal urethritis. The latest results show that there are a few lubricants with antimicrobiotic influence on MRSA (methicillin-resistant Staphylococcus aureus), which is of great importance because of the steady increase in MRSA dependent infections. PMID- 15806343 TI - A linkage map of chickpea (Cicer arietinum L.) based on populations from Kabuli x Desi crosses: location of genes for resistance to fusarium wilt race 0. AB - Two recombinant inbred line (RIL) populations derived from intraspecific crosses with a common parental line (JG62) were employed to develop a chickpea genetic map. Molecular markers, flower colour, double podding, seed coat thickness and resistance to fusarium wilt race 0 (FOC-0) were included in the study. Joint segregation analysis involved a total of 160 markers and 159 RILs. Ten linkage groups (LGs) were obtained that included morphological markers and 134 molecular markers (3 ISSRs, 13 STMSs and 118 RAPDs). Flower colour (B/b) and seed coat thickness (Tt/tt) appeared to be linked to STMS (GAA47). The single-/double podding locus was located on LG9 jointly with two RAPD markers and STMS TA80. LG3 included a gene for resistance to FOC-0 (Foc0(1)/foc0(1)) flanked by RAPD marker OPJ20(600) and STMS marker TR59. The association of this LG with FOC-0 resistance was confirmed by QTL analysis in the CA2139 x JG62 RIL population where two genes were involved in the resistance reaction. The STMS markers enabled comparison of LGs with preceding maps. PMID- 15806344 TI - Association analysis of candidate genes for maysin and chlorogenic acid accumulation in maize silks. AB - Two compounds, the C-glycosyl flavone maysin and the phenylpropanoid product chlorogenic acid (CGA), have been implicated in corn earworm (Helicoverpa zea Boddie) resistance in maize (Zea mays L.). Previous quantitative trait locus (QTL) analyses identified the pericarp color (p) locus, which encodes a transcription factor, as the major QTL for maysin and CGA. QTL analysis has also implicated the dihydroflavanol reductase (DFR; E.C. no. 1.1.1.219) locus anthocyaninless1 (a1) and the duplicate chalcone synthase (CHS; E.C. no. 2.3.1.74) loci colorless2 (c2) and white pollen1 (whp1) as genes underlying QTL for maysin and/or CGA synthesis. Epistatic interactions between p and a1 and between p and c2 were also defined. CHS catalyzes the first step in the flavonoid pathway and represents one of the first enzyme steps following the branch off the general phenylpropanoid pathway towards CGA synthesis. In maize, the reduction of dihydroflavanol to leucoanthocyanin by DFR immediately follows the pathway branch leading to C-glycosyl flavone production. The detection of QTLs for maysin and CGA concentration at loci encoding enzyme steps following the pathway branch points implicates alterations in the flow of biochemical intermediates as the biological basis of the QTL effects. To examine if sequence variation among alleles of a1, c2, and whp1 affect maysin and CGA synthesis in maize silks, we performed an association analysis. Because the p locus has often been a major QTL for maysin and CGA and has exhibited epistatic interactions with a1, c2, and whp1, association analysis was conditioned on the p genotype. A highly significant association of two sequence polymorphisms in the promoter of a1 with maysin synthesis was demonstrated. Additional conditioning on the genotype of the significant a1 polymorphism allowed the detection of a significant polymorphism within the whp1 promoter. Our analyses demonstrate that conditioning for epistatic factors greatly increases the power of association testing. PMID- 15806345 TI - Genetic relationships within Brassica rapa as inferred from AFLP fingerprints. AB - Amplified fragment length polymorphism (AFLP) markers were employed to assess the genetic diversity amongst two large collections of Brassica rapa accessions. Collection A consisted of 161 B. rapa accessions representing different morphotypes among the cultivated B. rapa, including traditional and modern cultivars and breeding materials from geographical locations from all over the world and two Brassica napus accessions. Collection B consisted of 96 accessions, representing mainly leafy vegetable types cultivated in China. On the basis of the AFLP data obtained, we constructed phenetic trees using MEGA 2.1: software. The level of polymorphism was very high, and it was evident that the amount of genetic variation present within the groups was often comparable to the variation between the different cultivar groups. Cluster analysis revealed groups, often with low bootstrap values, which coincided with cultivar groups. The most interesting information revealed by the phenetic trees was that different morphotypes are often more related to other morphotypes from the same region (East Asia vs. Europe) than to similar morphotypes from different regions, suggesting either an independent origin and or a long and separate domestication and breeding history in both regions. PMID- 15806346 TI - Reproduction and cytogenetic characterization of interspecific hybrids derived from crosses between Brassica carinata and B. rapa. AB - The tri-genomic hybrid (ABC, 2n=27) between Brassica carinata (BBCC, 2n=34) and B. rapa (AA, 2n=20) is a unique material for studying genome relationships among Brassica species and a valuable bridge for transferring desirable characteristics from one species to the other within the genus Brassica. The crossability between B. carinata and B. rapa was varied with the cultivar of B. rapa. Hybrid pollen mother cells (PMCs), confirmed by morphological observation and molecular marker assay, could be grouped into 20 classes on the basis of chromosome pairing configurations. More than 30% of the PMCs had nine or more bivalents. Genomic in situ hybridization confirmed that two of the bivalents most likely belonged to the B genome. Nearly one-half of the PMCs had trivalents (0-2) and quadrivalents (0-2), which revealed partial homology among the A, B, and C genomes and suggested that there is a good possibility to transfer genes by means of recombination among the three genomes. The advantages of using the tri-genomic hybrids as bridge material for breeding new types of B. napus are discussed. PMID- 15806347 TI - Methods for predicting superior genotypes under multiple environments based on QTL effects. AB - Methods were developed for predicting two kinds of superior genotypes (superior line and superior hybrid) based on quantative trait locus (QTL) effects including epistatic and QTL x environment interaction effects. Formulae were derived for predicting the total genetic effect of any individual with known QTLs genotype derived from the mapping population in a specific environment. Two algorithms, enumeration algorithm and stepwise tuning algorithm, were used to select the best multi-locus combination of all the putative QTLs. Grain weight per plant (GW) in rice was analyzed as a working example to demonstrate the proposed methods. Results showed that the predicted superior lines and superior hybrids had great superiorities over the F(1) hybrid, indicating large breeding potential remained for further improvement on GW. Results also showed that epistatic effects and their interaction with environments largely contributed to the superiorities of the predicted superior lines and superior hybrids. User-friendly software, QTLNetwork, version 1.0, was developed based on the methods in the present paper. PMID- 15806348 TI - A microarray analysis of wheat grain hardness. AB - Grain hardness is an important quality characteristic of wheat grain, and considerable research effort has focused on characterising the genetic and biochemical basis underlying the hardness phenotype. Previous research has shown that the predominant difference between hard and soft seeds is linked to the puroindoline (PIN) proteins. In this study the near-isogenic lines of Heron and Falcon, which differ only in the grain hardness character, were compared using a cDNA microarray consisting of approximately 5,000 unique cDNA clones that were isolated from wheat and barley endosperm tissue. Our analysis showed that major differences in gene expression were evident for puroindoline-a (Pina), with a minor but not consistent change in the expression of puroindoline-b (Pinb). These observations were confirmed using a 16,000 unique cDNA microarray in a comparison of hard wheats with either the Pina null or Pinb mutation. PMID- 15806349 TI - Cross-species amplification of Medicago truncatula microsatellites across three major pulse crops. AB - Model plants are facilitating the genetic characterization and comparative mapping of a number of traditional crops. Medicago truncatula has been widely accepted as a model plant to this end as it provides the essential tools for multiple aspects of legume genetics and genomics. A large set of markers from highly conserved M. truncatula gene regions is being created and used to establish a worldwide framework for comparative genomic studies in legumes. We have investigated the potential for cross-species amplification of 209 expressed sequence tag (EST)-based and 33 bacterial artificial chromosome (BAC)-based microsatellites from M. truncatula in the three most important European legume pulses-pea, faba bean and chickpea-that might facilitate future comparative mapping. Our results revealed significant transferability of M. truncatula microsatellites to the three pulses (40% in faba bean, 36.3% in chickpea and 37.6% in pea). The percentage of M. truncatula EST-SSRs (simple sequence repeats) amplified in the three crops (39-43%) was twofold higher than that of the genomic SSRs (21-24%). Sequence analysis determined that the level of conservation in the microsatellite motif was very low, while the flanking regions were generally well conserved. The variations in the sequences were mainly due to changes in the number of repeat motifs in the microsatellite region combined with indel and base substitutions. None of the functional microsatellites showed direct polymorphism among the parental genotypes tested, consequently preventing their immediate use for mapping purposes. PMID- 15806350 TI - Intersubgenomic heterosis in seed yield potential observed in a new type of Brassica napus introgressed with partial Brassica rapa genome. AB - This paper reports the observation on the intersubgenomic heterosis for seed yield among hybrids between natural Brassica napus (A(n)A(n)C(n)C(n)) and a new type of B. napus with introgressions of genomic components of Brassica rapa (A(r)A(r)). This B. napus was selected from the progeny of B. napus x B. rapa and (B. napus x B. rapa) x B. rapa based on extensive phenotypic and cytological observation. Among the 129 studied partial intersubgenomic hybrids, which were obtained by randomly crossing 13 lines of the new type of B. napus in F(3) or BC(1)F(3) to 27 cultivars of B. napus from different regions as tester lines, about 90% of combinations exceeded the yield of their respective tester lines, whereas about 75% and 25% of combinations surpassed two elite Chinese cultivars, respectively. This strong heterosis was further confirmed by reevaluating 2 out of the 129 combinations in a successive year and by surveying hybrids between 20 lines of the new type of B. napus in BC(1)F(5) and its parental B. napus in two locations. Some DNA segments from B. rapa were identified with significant effects on seed yield and yield components of the new type of B. napus in BC(1)F(5) and intersubgenomic hybrids in positive or negative direction. It seems that the genomic components introgressed from B. rapa contributed to improvement of seed yield of rapeseed. PMID- 15806352 TI - Chromatography in silico, basic concept in reversed-phase liquid chromatography. AB - Basic phenomena in reversed-phase liquid chromatography have been quantitatively analyzed using a computational chemical calculation. Pyridine interacted with an ionized silica surface under neutral conditions. Alkyl-chain length affected the contact surface area with an analyte. Steric hindrance was demonstrated using a model graphitic carbon phase and unsaturated alkenes. Quantitative structure retention relationships in reversed-phase liquid chromatography were demonstrated for phenolic compounds and acidic and basic drugs. The correlations between predicted and measured retention factors were satisfactory. Dissociation constants were derived from the atom partial charge and used to predict retention factors of partially ionized compounds. PMID- 15806353 TI - Antimicrobial activity of conditioned medium fractions from Spodoptera frugiperda Sf9 and Trichoplusia ni Hi5 insect cells. AB - Concentrated conditioned medium (CM) fractions from Spodoptera frugiperda Sf9 and Trichoplusia ni cells, eluting from a gel filtration column at around 10 kDa, were found to exhibit strong antibacterial activity against Bacillus megaterium and Escherichia coli. The B. megaterium cells incubated in the CM fraction from Sf9 cells rapidly lost viability: after 8 min the viability had decreased to 0.7%, as compared with the control. Addition of the CM fraction to E. coli cells resulted in a less drastic drop in viability: 65% viability was lost after 60 min of incubation. Further, exposure to the CM fraction caused a substantial leakage of intracellular proteins, as demonstrated by SDS-PAGE analysis. Cell lysis was confirmed by optical density measurements, microscopic investigations and flow cytometry. B. megaterium exposed to a CM fraction from T. ni cells lost 97% of their viability in about 40 min. Ubiquitin, thioredoxin and cyclophilin were identified in the antibacterial fraction from Sf9 cells by mass spectrometry and N-terminal amino acid sequencing. Other proteins in the fraction gave no matches in a database search. Since ubiquitin was shown not to cause the antimicrobial effect and thioredoxin and cyclophilin were likely not involved, the responsible agent may be an unknown protein, not yet registered in databases. The antimicrobial effect of the CM fraction from T. ni cells most probably comes from a lysozyme precursor protein. PMID- 15806354 TI - Microbial response and elimination capacity in biofilters subjected to high toluene loadings. AB - Elimination capacity (EC) is frequently used as a performance and design criterion for vapor-phase biofilters without further verification of the microbial quantity and activity. This study was conducted to investigate how biofilters respond to high pollutant loadings and ultimately how this affects the EC of the biofilter. Two identical laboratory-scale biofilters were maintained at an initial toluene loading rate of 46 g m-3 h-1 for a period of 24 days. After the initial biofilm development stage, the loading rates were increased to 91 g m 3 h-1 and 137 g m-3 h-1, respectively. Following a short period of pseudo-steady state, toluene removal efficiencies rapidly declined in both biofilters, with a concurrent decline in both critical and maximum ECs. The decline was mainly due to deterioration in the biodegradation activity of the biofilm and a decline in the toluene-degrading bacterial population within the biofilm phase. The findings imply that high toluene loadings accelerated the deterioration in overall performance due to a rapid accumulation of inactive biomass. As a result, care must be used when relying on EC values for biofilter design and operational purposes, since the values do not appropriately reflect the temporal changes in biodegradation activity and active biomass quantities that can occur in biofilters subjected to high inlet loadings. PMID- 15806355 TI - Eco-Efficiency Analysis of biotechnological processes. AB - Eco-Efficiency has been variously defined and analytically implemented by several workers. In most cases, Eco-Efficiency is taken to mean the ecological optimization of overall systems while not disregarding economic factors. Eco Efficiency should increase the positive ecological performance of a commercial company in relation to economic value creation--or to reduce negative effects. Several companies use Eco-Efficiency Analysis for decision-making processes; and industrial examples of best practices in developing and implementing Eco Efficiency have been reviewed. They clearly demonstrate the environmental and business benefits of Eco-Efficiency. An instrument for the early recognition and systematic detection of economic and environmental opportunities and risks for production processes in the chemical industry began use in 1997, since when different new features have been developed, leading to many examples. This powerful Eco-Efficiency Analysis allows a feasibility evaluation of existing and future business activities and is applied by BASF. In many cases, decision-makers are able to choose among alternative processes for making a product. PMID- 15806356 TI - A microcosm study on bioremediation of p-nitrophenol-contaminated soil using Arthrobacter protophormiae RKJ100. AB - p-Nitrophenol (PNP), a toxic nitroaromatic compound, can build up in soils due to extensive usage of nitrophenolic pesticides and hence needs to be removed. Arthrobacter protophormiae RKJ100, a PNP-degrading organism, was used in this work to study factors affecting its growth, and then evaluated for its capacity to degrade PNP in soil microcosms. Molasses (10%) treated with 0.1% potassium hexacyanoferrate was found to be a suitable and cheap carbon source for inoculum preparation. Induction studies showed that PNP depletion was quicker when cells were induced by pre-exposure to PNP. The efficiency of PNP degradation in soil by strain RKJ100 was seen to be dependent on pH, temperature, initial PNP concentration and inoculum size. Microcosm studies performed with varying concentrations (1.4-210 ppm) of PNP-spiked soils showed that strain RKJ100 could effectively degrade PNP over the range 1.4-140 ppm. A cell density of 2x10(8) colony forming units/g soil was found to be suitable for PNP degradation over a temperature range of 20-40 degrees C and at a slightly alkaline pH (7.5). Our results indicate that strain RKJ100 has potential for use in in situ bioremediation of PNP-contaminated sites. This is a model study that could be used for decontamination of sites contaminated also with other compounds. PMID- 15806357 TI - Reverse oblique intertrochanteric femoral fractures treated with the gamma nail. AB - We studied 47 reverse oblique intertrochanteric femoral fractures that were treated with gamma nails between 1992 and 2000. Fracture reduction was satisfactory in 38 patients (81%), the hip screw position was correct in 42 (89%) and there were no peri-operative complications. However, in three patients the nail displaced, resulting in non-union in one and protrusion into the acetabulum in another. A logistical regression analysis of our series showed that an incorrect position of the hip screw in the femoral head was the only predictor for complications. Thus, we consider that the gamma nail is a good option for the treatment of these complex fractures. PMID- 15806358 TI - Peri-acetabular radiolucent lines: inter- and intra-observer agreement on post operative radiographs. AB - Peri-acetabular radiolucent lines (RLLs) seen on "early" post-operative radiographs have been identified as a potential predictor of long-term implant performance. This study examines the inter- and intra-observer variation encountered when assessing such radiographs. Four consultant orthopaedic surgeons assessed the presence, extent and width of RLLs in 220 radiographs performed on 50 patients taken one to two weeks, six weeks, six months and one year following surgery. Inter-observer agreement was fair at 7-14 days but improved to moderate to good in films at six and 12 months. Intra-observer agreement was moderate to good at 7-10 days but again improved to good at 6 and 12 months. When only the presence or absence of RLLs was considered, both inter-observer and intra observer agreement improved for both the six-month and one-year radiographs. This experiment shows that caution must be used for the interpretation of RLLs on hip radiographs taken during the very early post-operative period. We recommend that films taken at least six weeks to six months following surgery should be used for assessment to reduce observer variation. For optimum results, a single experienced observer should do the assessment with a simple classification. PMID- 15806359 TI - The incidence of septic arthritis in Malawian children. AB - The incidence of septic arthritis in Malawian children was estimated by assessing the incidence in the relatively enclosed district of Chikwawa. We found an incidence of approximately one in 5,000 in the under-five age group and one in 13,000 in the 5-15 age group. PMID- 15806360 TI - Anterior thigh pain after cementless total hip arthroplasty. AB - We compared the incidence of anterior thigh pain retrospectively following primary uncemented total hip replacement in 70 patients (78 hips) with osteoarthritis. In one group (38 hips), an Endoplus porous-coated, canal-filling prosthesis was inserted through an anterolateral approach. In the other group (40 hips), a Furlong hydroxyapatite-coated prosthesis was inserted through a posterior approach. The latter prosthesis has a relatively longer femoral stem and does not achieve diaphyseal press-fit contact. Three patients in the Furlong group had anterior thigh pain, which eventually resolved in two patients. In the Endoplus group, two patients developed anterior thigh pain; in both cases, there was an underlying cause, which had to be treated. Idiopathic anterior thigh pain is more common in non-canal-filling prostheses. When anterior thigh pain is noticed in canal-filling prosthesis, a search for and treatment of the cause will cure the pain. The hydroxyapatite coating did not prevent thigh pain in the Furlong group. PMID- 15806361 TI - Abdominal tuberculous lymphadenopathy: MRI features. AB - The aim of this study was to describe the MRI features of abdominal tuberculous lymphadenopathy. MRI studies of 13 patients with abdominal tuberculous lymphadenopathy were reviewed with regard to anatomic distribution and size. Signal intensities, in relation to abdominal wall muscle, on unenhanced T1- and T2-weighted images and patterns of contrast enhancement of lymphadenopathy were evaluated in each patient. In each patient, the largest lymph node with the same imaging characteristic was evaluated. The upper paraaortic region was the most common site of involvement (n=12 patients), followed by the lesser omentum (n=10 patients), the anterior pararenal space (n=9 patients), the lower paraaortic area (n=8 patients), the small bowel mesentery (n=6 patients), the greater omentum (n=2 patients) and the originating site of the inferior mesenteric artery (n=2 patients). The mean lymph node size was 1.8 cm (range 0.5-5 cm). The overall mean lymph node number per patient was 16 (range 2-50). A total of 41 lymph nodes were evaluated in 13 patients. On T2-weighted images, 40 lesions were hyperintense and one lesion was isointense. Nine hyperintense lesions showed a hypointense peripheral rim and seven internal heterogeneity. Perinodal T2-hyperintensity was present in 23 lesions. The latter finding was valid for all patients. On T1 weighted images, 30 lesions were hypointense and 11 isointense. Nine hypointense lesions demonstrated a hyperintense peripheral rim, and six were heterogeneous. Contrast-enhanced fat-suppressed T1-weighted images demonstrated predominant peripheral enhancement in 28 lesions: (1) peripheral uniform, thin (n=19); (2) thick irregular, complete (n=3); and (3) conglomerate group of nodes showing peripheral and central areas of rim enhancement (n=6). Heterogeneous and homogeneous enhancement was present in ten and three lesions, respectively. Combinations of enhancing patterns in the same nodal group and different nodal groups were seen in eight and nine patients, respectively. Abdominal tuberculous lymphadenopathy may show a variety of signal intensities and patterns of contrast enhancement on MRI. Lymphadenopathy, hypointense on T1-weighted, hyperintense on T2-weighted images with perinodal hyperintensity, and predominant peripheral rimlike enhancement may suggest the diagnosis of tuberculosis. PMID- 15806362 TI - Predictions of outcomes of renal stones after extracorporeal shock wave lithotripsy from stone characteristics determined by unenhanced helical computed tomography: a multivariate analysis. AB - The aim of our study is to analyze the relationships between the characteristics of renal stones determined by unenhanced helical computed tomography (UHCT) and their outcomes after extracorporeal shock wave lithotripsy (ESWL) as well as to predict ESWL outcomes of renal stones by their UHCT characteristics with the use of multivariate analysis. During a 7-month period, 80 adult patients with renal stones underwent ESWL as well as UHCT both before and 3 months after ESWL. Of the 80 patients, 42 patients were classified as ESWL successes and 38 as ESWL failures based on their post-ESWL UHCT findings. For pre-ESWL UHCT, a stone number of more than 2 (P=0.0236), a maximal stone size of greater than 12 mm (P<0.0001), a stone burden of more than 700 mm3 (P<0.0001), a maximal stone density of more than 900 HU (P=0.0008) and nonround/oval stones (P=0.0007) were associated with ESWL failure outcomes. Multivariate analysis demonstrated that a stone burden of more than 700 mm3 (P=0.0003), the presence of nonround/oval stones (P=0.0072) and a maximal stone density of more than 900 HU (P=0.0430) were statistically significant predictors of a failure outcome for ESWL. Thus, the analysis of stone characteristics of renal stones by UHCT is helpful in selecting appropriate patients undergoing ESWL for favorable outcomes and reduces the overall costs of the treatment of renal stones. PMID- 15806363 TI - Flat detectors and their clinical applications. AB - Diagnostic and interventional flat detector X-ray systems are penetrating the market in all application segments. First introduced in radiography and mammography, they have conquered cardiac and general angiography and are getting increasing attention in fluoroscopy. Two flat detector technologies prevail. The dominating method is based on an indirect X-ray conversion process, using cesium iodide scintillators. It offers considerable advantages in radiography, angiography and fluoroscopy. The other method employs a direct converter such as selenium which is particularly suitable for mammography. Both flat detector technologies are based on amorphous silicon active pixel matrices. Flat detectors facilitate the clinical workflow in radiographic rooms, foster improved image quality and provide the potential to reduce dose. This added value is based on their large dynamic range, their high sensitivity to X-rays and the instant availability of the image. Advanced image processing is instrumental in these improvements and expand the range of conventional diagnostic methods. In angiography and fluoroscopy the transition from image intensifiers to flat detectors is facilitated by ample advantages they offer, such as distortion-free images, excellent coarse contrast, large dynamic range and high X-ray sensitivity. These characteristics and their compatibility with strong magnetic fields are the basis for improved diagnostic methods and innovative interventional applications. PMID- 15806364 TI - Development of subdural effusions in association with pyruvate dehydrogenase deficiency. PMID- 15806365 TI - Liver metastasis with apparent intratumoral superparamagnetic iron oxide uptake. PMID- 15806366 TI - Multislice CT of the pelvis: dose reduction with regard to image quality using 16 row CT. AB - To optimize examination protocols of 16-row multi-detector CT (MDCT) of pelvis for dose reduction with regard to image quality. MDCT of pelvis was performed on 12 cadaver specimens with stepwise reduction of tube current from 160 mA (113, 80, 56, 40, 28) to 20 mA at 120 kV. Scan parameters were 16 x 1.5 mm collimation. Reconstructions of axial and coronal images were used for evaluation of cortex, trabeculum, image quality, image noise, acetabulum and iliosacral (ISJ) joints. After data were blinded, evaluation of images was done by three radiologists according to 5-point Likert scale. Accuracy of the observers in sorting films according to dose reduction was determined with kappa coefficient. Mean values of image evaluation were determined. Pronounced deterioration of image quality for all criteria was observed between 80 and 28 mA. Adequate image quality was obtained at 40 mA [effective dose (E): 2.2 mSv, CTDI(w): 2.8 mGy] for criterion detailed definition of acetabulum and ISJ and at 80 mA (E: 4.4 mSv, CTDI(w): 5.6 mGy) for remaining criteria. Moderate agreement was observed between the three observers (kappa coefficient: 0.31). All observers were excellent in arranging images according to decreasing dose. Using 16-row MDCT image quality of pelvis is acceptable at 80 mA and 120 kV. This translates into a dose reduction of 33% of average value of the nationwide survey of the German Roentgen Society (1999) for this type of examination. PMID- 15806367 TI - Mammary fibromatosis mimicking recurrent breast cancer: radiological findings. PMID- 15806368 TI - In vitro characterization of atherosclerotic carotid plaque with multidetector computed tomography and histopathological correlation. AB - This in vitro study evaluated the performance of 16-slice multidetector computed tomography (MDCT) in the assessment of carotid plaque components, with histology as the gold standard. Twenty-one specimens (n=21) were scanned and reconstructed after optimization of the protocol. Three corresponding MDCT images and histologic sections were selected from each specimen. The Hounsfield values (HV) of the major plaque components (calcifications, fibrous tissue and lipid) were assessed. Plaque areas (mm2) assessed with MDCT were compared with the results from histologic analysis. A value of 140 kVp and an intermediate reconstruction algorithm was the optimal protocol. In 15 out of 21 specimens it was possible to match MDCT images with histology. The HV of calcifications, fibrous tissue and lipid were 45+/-21, 79+/-20 and 960+/-491 HU (P<0.001), respectively. Plaque areas were compared in 27 matched levels. The calcified and lipid areas on MDCT and histology correlate well (R2=0.83 and R2=0.68, respectively). The mean difference in lipid area was 0.1 mm2 (95% CI=-2.1-2.3 mm2). This in vitro study showed that MDCT is capable of characterizing and quantifying the lipid rich portion of the atherosclerotic plaque. PMID- 15806369 TI - Comparison of CT, MRI and FDG-PET in response prediction of patients with locally advanced rectal cancer after multimodal preoperative therapy: is there a benefit in using functional imaging? AB - The aim of this study was to compare CT, MRI and FDG-PET in the prediction of outcome of neoadjuvant radiochemotherapy in patients with locally advanced primary rectal cancer. A total of 23 patients with T3/4 rectal cancer underwent a preoperative radiochemotherapy combined with regional hyperthermia. Staging was performed using four-slice CT (n=23), 1.5-T MRI (n=10), and (18)F-FDG-PET (n=23) before and 2-4 weeks after completion of neoadjuvant treatment. Response criteria were a change in T category and tumour volume for CT and MRI and a change in glucose uptake (standard uptake value) within the tumour for FDG-PET. Imaging results were compared with those of pretherapy endorectal ultrasound and histopathological findings. Histopathology showed a response to neoadjuvant therapy in 13 patients whereas 10 patients were classified as nonresponders. The mean SUV reduction in responders (60+/-14%) was significantly higher than in nonresponders (37+/-31%; P=0.030). The sensitivity and specificity of FDG-PET in identifying response was 100% (CT 54%, MRI 71%) and 60% (CT 80%, MRT 67%). Positive and negative predictive values were 77% (CT 78%, MRI 83%) and 100% (CT 57%, MRI 50%) (PET P=0.002, CT P=0.197, MRI P=0.500). These results suggest that FDG-PET is superior to CT and MRI in predicting response to preoperative multimodal treatment of locally advanced primary rectal cancer. PMID- 15806371 TI - Vector analysis of high early postoperative astigmatism after congenital cataract surgery. AB - BACKGROUND: Corneal astigmatism is a well-documented finding after cataract surgery. Postoperative astigmatism is of greater importance in children than in adults, because of its adverse effect on vision development and the risk of amblyopia. The purpose of this study was to evaluate refractive outcomes in eyes- which had high early postoperative astigmatism after congenital cataract surgery. METHODS: We retrospectively reviewed the charts of 74 children (112 eyes), who underwent congenital cataract extraction with intraocular lens implantation, using limbal incision, scleral tunnel, or clear corneal incision. Thirty-two eyes of 28 children, aged 2 months to 11 years, had astigmatism of 3.0 diopters (D) or more when assessed 1 week after surgery. Changes in cylinder vector, spherical equivalent (SEQ) and defocus equivalent (DEQ) between 1 week and 5 months postoperatively were calculated for each group. RESULTS: Mean SEQ and DEQ were 0.7+/-3.5 D and 5.1+/-2.8 D, respectively, at 1 week postoperatively- and -0.2+/ 3.4 D and 3.1+/-2.3 D, respectively, at 5 months postoperatively in the three groups. DEQ levels showed a significant reduction during the 5-month follow-up period (P<0.001). The mean astigmatism vector was 2.9+/-3.3 D x 93 degrees in all the three groups 1 week postoperatively- and 0.2+/-1.1 D x 153 degrees at 5 months after surgery. A significant change during 5-month follow-up in cylinder vector (P<0.02) and SEQ (P=0.01) was found after operations using scleral tunnel technique. CONCLUSION: Children- who underwent congenital cataract surgery by different surgical techniques showed a significant spontaneous reduction in DEQ values. A significant change in cylinder vector and SEQ was found after operations using scleral tunnel technique. PMID- 15806370 TI - [Ocular allergies]. AB - Recent developments indicate that ocular allergy is more than an IgE-mediated allergic conjunctivitis. Ocular allergy is a disease affecting the entire ocular surface including conjunctiva, lids, cornea, lacrimal gland and tear film. Besides an IgE-mediated reaction, a complex chronic inflammation is involved in the pathogenesis of many ocular allergies. According to their pathogenesis and clinical picture, ocular allergies are classified into mild forms, such as seasonal and perennial allergic conjunctivitis as well as giant papillary conjunctivitis, and chronic, potentially blinding forms such as atopic keratoconjunctivitis and vernal keratoconjunctivitis. New therapeutics act on the entire inflammatory process or try to modulate the allergic reaction early and specifically. The association with non-ocular allergic symptoms requires an interdisciplinary approach. PMID- 15806372 TI - Primary intraocular lymphoma of T-cell type: report of a case and review of the literature. AB - PURPOSE: Primary intraocular lymphoma (PIOL) is an uncommon non-Hodgkin lymphoma and is usually of B-cell type. Intraocular T-cell or T/NK-cell lymphomas are extremely rare and mostly represent a secondary manifestation of either a cutaneous or a systemic lymphoma. The aim of the current paper is to report the clinical, histopathological and molecular biological findings of a PIOL of T-cell type. METHODS: Conventional cytological and immunocytological examination of vitrectomy specimens. Conventional histology, immunohistochemistry and polymerase chain reaction (PCR) for the detection of immunoglobulin heavy chain (IgH) and T cell-receptor gamma (TCR-gamma) gene rearrangement, GeneScan analysis, and DNA sequencing were performed on the chorioretinal biopsy. RESULTS: Cytology of the right vitreous aspirate revealed a moderate cellular infiltrate consisting of medium-sized T-cells with pleomorphic nuclei. Similar atypical lymphocytes were seen in the partially necrotic chorioretinal biopsy. These lymphocytes expressed CD3, CD4, betaF1 and CD30, with a growth fraction of 90%. TCR-gamma-PCR, GeneScan analysis and DNA sequencing demonstrated a monoclonal amplification product within the expected range. In contrast, IgH-PCR revealed oligoclonal amplificates. The patient was treated with low-dose radiotherapy (total 45 Gy), and was in complete remission at final follow-up. CONCLUSION: A rare PIOL of T cell type was diagnosed on the basis of vitreous aspiration and chorioretinal biopsy. In addition to conventional cytology and immunocytology, the utilisation of gene rearrangement studies on vitreous or chorioretinal biopsies increases the chances of diagnosing or excluding a PIOL of either B-cell or T-cell type. Despite its rarity, ophthalmic pathologists should always consider the diagnosis of T-PIOL when reviewing vitreous samples. PMID- 15806373 TI - Fibrous proliferation of the pre-papillary canal in proliferative diabetic retinopathy: Cloquet's canal as a scaffold for proliferative diabetic retinopathy. AB - BACKGROUND: The structure of the vitreous body is known to play an important role in the pathogenesis of proliferative diabetic retinopathy (PDR). The purpose of this study was to ascertain the pattern of fibrous membrane proliferation in PDR focusing on pre-optic disc canal-like fibrous proliferation. METHODS: The investigation comprised two parts. The first, prospective study involved examining ten eyes of ten patients with PDR who underwent triamcinolone acetonide assisted pars plana vitrectomy (TA-PPV). The presence of canal-like intravitreous structures was determined during TA-PPV. The second, retrospective study included analysis of photographs and clinical records of 235 eyes examined in 207 patients with PDR. The clinical and surgical records of the 235 eyes treated with PPV were reviewed and were categorized into four groups based upon the pattern of fibrous proliferation (optic disc type, pre-cortical pocket type, mixed type, or other). Each clinical feature was studied. The clinical features of the four groups of PDR were compared. RESULTS: A pre-optic disc canal-like structure was observed in seven of the ten eyes. In the retrospective study, 25 eyes (10.7%) were classified as optic disc type, 16 eyes (6.9%) as pre-cortical pocket type, 68 eyes (29.1%) as mixed type, and 126 eyes (53.4%) as other types. The patients with the optic disc type were significantly younger than the other three groups (P = 0.02). Other clinical features and post-operative outcome were not significantly different between the groups. CONCLUSIONS: A pre-optic disc canal like structure, probably Cloquet's canal and the area of Martegiani, was present in PDR eyes. The PDR patients with fibrous proliferation of this structure were younger than those with other patterns of fibrous proliferation. PMID- 15806374 TI - Integrated visual fields: a new approach to measuring the binocular field of view and visual disability. AB - BACKGROUND: We have developed a method of quantifying the central binocular visual field by merging results from monocular fields (Integrated visual field). This study aims to compare the new measure with the binocular Esterman visual field test in identifying patients with self-reported visual disability. METHODS: Forty-eight patients with glaucoma each recorded Humphrey 24-2 fields for both eyes and an Esterman on the same day, and each completed a binary forced-choice questionnaire relating to perceived visual disability. Computer software merged sensitivity values from monocular fields to generate an integrated visual field and a related score of the number of defects at the <10 dB and <20 dB level. Receiver operating characteristic (ROC) analysis was used to compare the integrated visual field score and the Esterman disability score with individual responses to the questions on perceived difficulty with visual tasks. RESULTS: Comparison of areas under ROC curves revealed that a score based on the integrated visual field was generally better (median area: 0.79) than Esterman scores (median area: 0.70) in classifying patients with or without a self reported perceived difficulty with visual tasks. CONCLUSIONS: The integrated visual field offers a rapid assessment of a glaucoma patient's binocular visual field without extra perimetric testing. As compared to an actual binocular field test (Esterman), the integrated visual field provides a better prediction of a glaucoma patient's perceived inability to perform certain visual tasks. PMID- 15806375 TI - Macular pigment density in age-related maculopathy. AB - PURPOSE: Macular pigment (MP) is believed to have a protective role in the development and progression of age-related maculopathy (ARM). We present results of measurements of macular pigment density (MPD) in patients with ARM at different stages with a modified confocal scanning laser ophthalmoscope (HRA, Heidelberg Engineering, Heidelberg, Germany). METHOD: ARM patients were categorized into four groups. Staging depended upon the size of drusen, geographic atrophy, and loss of visual acuity due to ARMD in the fellow eye; 1: drusen size <125 microm, 2: drusen size >125 microm, 3: drusen size <125 microm + ARMD in fellow eye, and 4: drusen size >125 microm + ARMD in fellow eye. We took autofluorescence images at 488 nm and 514 nm with a modified HRA. MP density was evaluated within 2 degrees around the center of the fovea. In this study, we included 146 patients with ARM: 26 in group 1, 19 in group 2, 80 in group 3, and 21 in group 4. RESULTS: The mean MPD of the different stages of ARM was: in group 1, 0.233 +/- 0.091 DU; group 2, 0.218 +/- 0.102 density units (DU); group 3, 0.252 +/- 0.085 DU; and group 4, 0.208 +/- 0.100 DU. Statistical analysis (Kruskal-Wallis test) showed no differences between groups. CONCLUSIONS: Recent studies in ARM did not result in significant differences of MPD between the various stages of ARM. Further longitudinal studies are requested to compare the incidence of ARMD in eyes with high and low MPD in order to provide definite evidence of the influence of MPD on the progression of ARMD. PMID- 15806376 TI - Bilateral anterior granulomatous uveitis associated with cutaneous Rosai-Dorfman disease. AB - BACKGROUND: Rosai-Dorfman disease (RDD) is a benign, rare, histiocytic disorder of unknown etiology. We describe the case of a 61-year-old woman who presented to the ophthalmology clinic with bilateral granulomatous anterior uveitis. METHODS: The uveitis was controlled on topical prednisolone acetate 1%. One month after presentation, she developed generalized macules and papules, and dermatologic and systemic evaluations were performed. RESULTS: On the basis of skin biopsy, purely cutaneous RDD was established. CONCLUSION: Anterior granulomatous uveitis may be the presenting sign of cutaneous RDD. Consultation is necessary for accurate diagnosis. PMID- 15806378 TI - Interdigitating dendritic cell tumor with breast and cervical lymph-node involvement: a case report and review of the literature. AB - Interdigitating dendritic cell tumor (IDCT) is an extremely rare malignancy. It occurs primarily in lymph nodes, but extranodal involvement has also been reported. A 38-year-old woman with IDCT with breast and cervical lymph-node involvement is reported in this paper. To our knowledge, this is the first case of IDCT originating from the breast. In the breast and lymph node, the tumor displayed diffuse sheets, fascicles and storiform growth pattern. It was composed of oval to spindle cells with pale to eosinophilic cytoplasm, ill-defined cell outlines, oval nuclei with vesicular chromatin and prominent eosinophilic nucleoli. Mitotic activity was three per ten high-power fields. The neoplastic cells were intermingled with small mature lymphocytes and plasma cells. Immunohistochemical studies showed that the tumor cells were strongly and diffusely positive for vimentin, CD68, S-100 protein, CD45/leukocyte common antigen and fascin and focally positive for lysozyme, alpha-1 antitrypsin and CD4. Ki-67 labeling index was 10%. The patient was treated with combined therapeutic approaches, including surgery, radiotherapy and chemotherapy. IDCT has the potential for an aggressive clinical course. However, 32 months after the initial diagnosis, the patient is still alive and being followed with a stable tumor burden. PMID- 15806379 TI - Dedifferentiated liposarcoma of the oral cavity with angiosarcomatous dedifferentiation. AB - We report a unique case of a 42-year-old woman with a dedifferentiated liposarcoma of the soft tissue of the oral cavity with angiosarcomatous dedifferentiation. Liposarcomas compromising the head and neck region are very unusual, and most of the cases in oral cavity show a well-differentiated pattern. Dedifferentiation in liposarcomas occurs in about 10% of the cases and, when it occurs, the dedifferentiated areas usually resemble high-grade fibrosarcoma or pleomorphic sarcoma. Divergent differentiation might also occur. To the best of our knowledge, there are only nine cases of dedifferentiated liposarcoma of the oral cavity, none of which show an angiosarcomatous dedifferentiation. PMID- 15806380 TI - Extracellular matrix proteins and matrix metalloproteinases differ between various right and left ventricular sites in end-stage cardiomyopathies. AB - This study was undertaken to investigate whether there might be differences in the distribution of extracellular matrix (ECM) proteins and matrix metalloproteinases (MMPs), depending on their specific sites within the heart. We investigated 33 explanted human hearts, 15 with dilated cardiomyopathy (DCM) and 18 with ischemic cardiomyopathy (ICM). Transmural samples from the right ventricle, the interventricular septum and the left ventricle, either from near the apex or from near the base were taken from every heart. Frozen sections were processed for connective tissue staining and immunohistochemistry for collagens type I, III, IV, laminin and fibronectin, as well as MMP-1, -2 and -9. Volume densities of laminin in ICM as well as of fibronectin and collagen types I and IV in DCM showed significant differences between right and left ventricular sites. The volume densities of matrix proteins usually did not reveal significant differences among the three left ventricular sites tested in both DCM and ICM. MMPs partly showed differences between the right and the left ventricular myocardium. These results suggest that the distributions of ECM proteins and MMPs differ between the two ventricles in both end-stage DCM and ICM. This gives rise to the hypothesis that a specific pattern of ECM degradation exists in the right and left ventricular myocardium. PMID- 15806381 TI - Ultrastructure and localization of a visual Gq protein in hypertrophied epitoke ocelli of Perinereis brevicirris (Polychaeta, Annelida). AB - Functional ultrastructural changes in the rhabdomeric photoreceptors of the cerebral ocelli are described for normal and sexually mature (epitoke) Perinereis brevicirris (Polychaeta, Annelida). With sexual maturation, the cerebral ocelli hypertrophied, increasing in volume to 5.5 times that of ocelli in the normal state, and the thickness of the retinal layer increased up to 10 times. Perinereis ocelli have a pigmented retinal layer consisting of at least two cell types: photoreceptor cell (PR) and pigmented supporting cells (PS). In epitoke ocelli, PR bear well-developed rhabdomeric microvilli, multilamellar bodies, and numerous cytoplasmic membranous structures, including vesicles, smooth endoplasmic reticulum, and secondary lysosomes. Localization of a visual Gq protein in the ocelli was studied with anti-GqC antibody. The antibody strongly labeled not only microvilli and multilamellar bodies throughout the retinal layer, but also secondary lysosomes and vesicles in the cytoplasm of the PR in the epitoke ocelli, although labeling was observed only in the microvilli and multilamellar bodies in normal ocelli. Reverse transcription/polymerase chain reaction analysis revealed that the amount of G protein alpha subunit mRNA in the epitoke head increased by roughly twice that of the normal head. Since Gq protein is essential for phototransduction in Perinereis ocelli, these results suggest that the sites are involved in photoreceptive membrane turnover, which occurs much more extensively in epitoke ocelli. Thus, epitoke ocelli may represent a model system for studying rhabdomeric photoreceptive membrane turnover. PMID- 15806382 TI - Improving performance of a gas stripping-based recovery system to remove butanol from Clostridium beijerinckii fermentation. AB - The effect of factors such as gas recycle rate, bubble size, presence of acetone, and ethanol in the solution/broth were investigated in order to remove butanol from model solution or fermentation broth (also called acetone butanol ethanol or ABE or solvents). Butanol (8 g L(-1), model solution, Fig. 2) stripping rate was found to be proportional to the gas recycle rate. In the bubble size range attempted (< 0.5 and 0.5-5.0 mm), the bubble size did not have any effect on butanol removal rate (Fig. 3, model solution). In Clostridium beijerinckii fermentation, ABE productivity was reduced from 0.47 g L(-1) h(-1) to 0.25 g L( 1) h(-1) when smaller (< 0.5 mm) bubble size was used to remove ABE (Fig. 4, results reported as butanol/ABE concentration). The productivity was reduced as a result of addition of an excessive amount of antifoam used to inhibit the production of foam caused by the smaller bubbles. This suggested that the fermentation was negatively affected by antifoam. PMID- 15806383 TI - Using a kinetic model that considers cell segregation to optimize hEGF expression in fed-batch cultures of recombinant Escherichia coli. AB - Growth inhibition of recombinant Escherichia coli during the expression of human epidermal growth factor was observed. The recombinant cells could be segregated into three populations based on their cell division and plasmid maintenance abilities: dividing and plasmid-bearing cells, dividing and plasmid-free cells, and viable-but-non-culturable (VBNC) cells. Fed-batch fermentations were performed to investigate the effect of cell segregation on the kinetics of growth and foreign protein production. The results showed that a low concentration of inducer caused weak induction, whereas high levels cause strong induction, resulting in cells segregating into VBNC bacteria and producing a low foreign protein yield. A kinetic model for cell segregation was proposed and its predictions correlated well with experimental data for cell growth and protein expression. The optimal induction strategy could then be predicted by the model, and this prediction was then verified by experimentally deriving the conditions necessary for maximum expression of recombinant protein. PMID- 15806384 TI - [Fibromyalgia -- a somatoform (pain) disorder?]. AB - The hypothesis that fibromyalgia (FM) should be classified as a somatoform disorder was assessed by reviewing current clinical studies. According to the ICD 10, somatic illness beliefs of the patient, high health care utilization, and frustrating patient-doctor relationships are diagnostic criteria of somatoform disorders. For the diagnosis of a somatoform pain disorder, a temporal association between the manifestation of pain and emotional or psychosocial conflicts and the exclusion of a depressive disorder are additionally required. Empirical studies demonstrate a higher lifetime and current prevalence of psychiatric disorders, childhood adversities, life events, and daily hassles and a higher health care utilization of FM patients. Studies also reveal that most patients believe that both somatic and psychosocial factors have caused their disorder. The patient-doctor relationship is characterized to be disappointing for both. Yet in all studies there were patients who did not fulfill the ICD-10 criteria of a somatoform (pain) disorder. A biopsychosocial model of FM differentiating between biological as well as psychosocial predisposing, triggering, and perpetuating factors in the pathogenesis of FM is presented as an alternative model. Hopefully the biopsychosocial model and the distinction of subgroups will enable more differentiated and tailored psychotherapeutic and pharmacological treatment strategies. PMID- 15806385 TI - [Alimentary trigger factors that provoke migraine and tension-type headache]. AB - Based on a review of the literature the authors discuss the role of nutrition in the precipitation of migraine and tension-type headache (TTH). The available information relies largely on the subjective assessment of the patients. Controlled trials suggest that alcohol and caffeine withdrawal are the most important nutritional precipitating factors of migraine and TTH. In addition, there is some evidence that missing meals is also an important factor. Dehydration seems to deserve more attention. A selective sensitivity to red wine has been shown in some patients, the importance of chocolate has been doubted seriously, and scientific evidence for cheese as a precipitating factor is lacking. Despite a series of experimental studies demonstrating that NO donors such as nitroglycerin and parenteral histamine cause headache the role of histamine, nitrates, and nitrites in food remains unclear. Similarly, other biogenic amines and aspartame have not been proven to precipitate headache. Sodium glutamate causes adverse reactions including headache probably at large doses ingested on an empty stomach. Therefore, patients should be advised that food plays a limited role as a precipitating factor of migraine and TTH. Subjective sensitivity to certain foods should be examined critically, and proven precipitating factors should be avoided. General dietary restrictions have not been proven to be useful. PMID- 15806386 TI - Synthesis of vanadium(IV,V) hydroxamic acid complexes and in vivo assessment of their insulin-like activity. AB - We synthesized vanadyl (oxidation state +IV) and vanadate (oxidation state +V) complexes with the same hydroxamic acid derivative ligand, and assessed their glucose-lowering activities in relation to the vanadium biodistribution behavior in streptozotocin-induced diabetic mice. When the mice received an intraperitoneal injection of the complexes, the vanadate complex more effectively lowered the elevated glucose levels compared with the vanadyl one. The glucose lowering effect of the vanadate complex was linearly related to its dose within the range from 2.5 to 7.5 mg V/kg. In addition, pretreatment of the vanadate complex induced a larger insulin-enhancing effect than the vanadyl complex. Both complexes were more effective than the corresponding inorganic vanadium compounds. The vanadyl and vanadate complexes, but not the inorganic vanadium compounds, resulted in almost the same organ vanadium distribution. Consequently, the observed differences in the insulin-like activity between the complexes would reflect the potency of the two compounds in the +IV and +V oxidation states in the subcellular region. PMID- 15806387 TI - Force field impact and spin-probe modeling in molecular dynamics simulations of spin-labeled T4 lysozyme. AB - Several attempts have been made to compute electron paramagnetic resonance (EPR) spectra of biomolecules, using motional models or simulated trajectories to describe dynamics. Ideally, the simulated trajectories should capture "fast" (picosecond) snapshots of spin-probe rotations accurately, while being lengthy enough to ensure a proper Fourier integration of the time-domain signal. It is the interplay of the two criteria that poses computational challenges to the method. In this context, an analysis of the spin-probe and protein conformational sampling and equilibration, with different force fields and with explicit solvent, may be a useful attempt. The present work reports a comparative study of the effect of the molecular dynamics (MD) force field on conformational sampling and equilibration in two spin-labeled T4 lysozyme (T4L) variants, N40C and K48C. Ensembles of 10x 3 ns-trajectories per variant and per force field (OPLS/AMBER and AMBER99) are analyzed for a reliable assessment of convergence and sampling. It is found that subtle site-dependent differences in spin-probe rotations and torsions are more readily captured in the AMBER99 trajectories than in the OPLS/AMBER simulations. On the other hand, sampling and equilibration are found to be better with the OPLS/AMBER force field at equal trajectory lengths. PMID- 15806388 TI - Computer simulation of polypeptide translocation through a nanopore. AB - A simplified model of polypeptide chains was designed and studied by means of computer simulations. Chains were represented by a sequence of united atoms located at the positions of the alpha-carbons. A further assumption was the lattice approximation for the chains. We used a (310) lattice, which was found useful for studying properties of proteins. The force field used consisted of a long-range contact potential between amino-acid residues and a local preference for forming alpha-helical states. The chain consisted of two kinds of residues: hydrophilic (P) and hydrophobic (H) ones forming model helical septets--HHPPHPP- in a sequence. The chains were placed near an impenetrable surface with a square hole in it. The size of the hole was comparable or smaller than the size of a chain. The properties of these model chains were determined using the Monte-Carlo simulation method. During the simulations, translocation of the chain through the hole in the wall was observed. The influence of the chain length, the temperature differences on both sides of the wall and the force field on the chain properties were investigated. It was shown that the translocation time scales as N(2.2) and it was found that the presence of the local helical potential significantly slows down the process of translocation. [Figure: see text]. The snapshots of typical chain's conformation obtained during the simulation for chain consisted of N = 60. The values of the local potential epsilon(loc) = -8. PMID- 15806390 TI - Improvement of bioremediation by Pseudomonas and Burkholderia by mutants of the Vitreoscilla hemoglobin gene (vgb) integrated into their chromosomes. AB - Using genetic engineering, the Vitreoscilla (bacterial) hemoglobin gene (vgb) was integrated stably into the chromosomes of Pseudomonas aeruginosa and Burkholderia sp. strain DNT. This was done for both wild type vgb and two site-directed mutants of vgb that produce Vitreoscilla hemoglobin (VHb) with lowered oxygen affinities; in all cases functional VHb was expressed. Similar to previous results, the wild type VHb improved growth for both species and degradation of 2,4-dinitrotoluene (Burkholderia sp.) or benzoic acid (P. aeruginosa) under both normal and low aeration conditions. Both mutant vgbs enhanced these parameters compared to wild type vgb, and the improvement was seen in both species. The enhancements were generally greater at low aeration than at normal aeration. The results demonstrate the possibility that the positive effects provided by VHb may be augmented by protein engineering. PMID- 15806391 TI - Comparing different controllers for the coordination of a six-legged walker. AB - This paper compares four models for coordination: the Cruse rules (Cruse et al. 199), two extended versions of a model based on ideas by Porta and Celaya (2001), and one model based on the idea of MMC networks (Steinkuhler and Cruse 1998). All models are capable of coordinating a six-legged walker on level terrain and of crossing low obstacles. The experiments presented here show that an MMC network for coordination is feasible and that the Porta variants, which are based on rather intuitive ideas, show the best performance. PMID- 15806392 TI - Background-activity-dependent properties of a network model for working memory that incorporates cellular bistability. AB - In models of working memory, transient stimuli are encoded by feature-selective persistent neural activity. Network models of working memory are also implicitly bistable. In the absence of a brief stimulus, only spontaneous, low-level, and presumably nonpatterned neural activity is seen. In many working-memory models, local recurrent excitation combined with long-range inhibition (Mexican hat coupling) can result in a network-induced, spatially localized persistent activity or "bump state" that coexists with a stable uniform state. There is now renewed interest in the concept that individual neurons might have some intrinsic ability to sustain persistent activity without recurrent network interactions. A recent visuospatial working-memory model (Camperi and Wang 1998) incorporates both intrinsic bistability of individual neurons within a firing rate network model and a single population of neurons on a ring with lateral inhibitory coupling. We have explored this model in more detail and have characterized the response properties with changes in background synaptic input I(o) and stimulus width. We find that only a small range of I(o) yields a working-memory-like coexistence of bump and uniform solutions that are both stable. There is a rather larger range where only the bump solution is stable that might correspond instead to a feature-selective long-term memory. Such a network therefore requires careful tuning to exhibit working-memory-like function. Interestingly, where bumps and uniform stable states coexist, we find a continuous family of stable bumps representing stimulus width. Thus, in the range of parameters corresponding to working memory, the model is capable of capturing a two-parameter family of stimulus features including both orientation and width. PMID- 15806393 TI - [Pharmacology of Sinupret. Recent results on the rational for the Sinupret compound]. AB - The herbal medicinal product Sinupret has been successfully used for the treatment of sinusitis for 70 years. In Germany the product is established as a standard in the therapy of sinusitis. The most recent pharmacodynamic findings resulted from close scientific cooperation with Pontus Stierna and co-workers from the Karolinska Institute in Stockholm: Mice were infected intranasally with Streptococcus pneumoniae to induce bacterial rhinosinusitis. Animals were randomized to treatment with ampicillin, dexamethasone, Sinupret, or sham treatment. All groups receiving active treatment showed a reduction in bacterial growth after 4 days and a significant reduction of bacterial growth after 8 days. Similar results were observed regarding histopathology. Initial findings in a sinusitis model in the New Zealand White rabbit confirmed the results observed in the aforementioned studies in mice. Understanding of the pharmacological profile of Sinupret supports the knowledge about the clinical efficacy of this herbal medicinal product. PMID- 15806394 TI - A homozygous nonsense mutation in SOX9 in the dominant disorder campomelic dysplasia: a case of mitotic gene conversion. AB - Campomelic dysplasia (CD; MIM 114290), an autosomal dominant skeletal malformation syndrome with XY sex reversal, is caused by heterozygous de novo mutations in and around the SOX9 gene on 17q. We report a patient with typical signs of CD, including sex reversal, who was, surprisingly, homozygous for the nonsense mutation Y440X. Since neither parent carried the Y440X mutation, possible mechanisms explaining the homozygous situation were a de novo mutation followed by uniparental isodisomy, somatic crossing over, or gene conversion. As the patient was heterozygous for six microsatellite markers flanking SOX9, uniparental isodisomy and somatic crossing over were excluded. Analysis of intragenic single-nucleotide polymorphisms suggested that the homozygous mutation arose by a mitotic gene conversion event involving exchange of at least 440 nucleotides and at most 2,208 nucleotides between a de novo mutant maternal allele and a wild-type paternal allele. Analysis of cloned alleles showed that homozygous mutant cells constituted about 80% of the leukocyte cell population of the patient, whereas about 20% were heterozygous mutant cells. Heterozygous Y440X mutations, previously described in three CD cases, have been identified in seven additional cases, thus constituting the most frequent recurrent mutations in SOX9. These patients frequently have a milder phenotype with longer survival, possibly because of the retention of some transactivation activity of the mutant protein on SOX9 target genes, as shown by cell transfection experiments. The fact that the patient survived for 3 months may thus be explained by homozygosity for a hypomorphic rather than a complete loss-of-function allele, in combination with somatic mosaicism. This is, to our knowledge, the first report of mitotic gene conversion of a wild-type allele by a de novo mutant allele in humans. PMID- 15806395 TI - AHSG gene variant is associated with leanness among Swedish men. AB - Alpha(2) Heremans-Schmid glycoprotein (AHSG) is a plasma protein inhibiting the activity of the insulin receptor tyrosine kinase. Ahsg knock-out mice have increased insulin sensitivity and are resistant to diet-induced obesity. We hypothesized that functional variants of the AHSG gene segregating in the human population would reflect variation in body mass index (BMI). We genotyped 356 overweight or obese (BMI: 37.2 [25.0-66.5] kg/m(2)) and 148 lean (BMI: 23.7 [23.4 24.9] kg/m(2)) otherwise healthy Swedish men for three non-synonymous single nucleotide polymorphisms (SNPs) within exon 6 (rs4917) and exon 7 (rs4918 and Arg299Cys) and one SNP in intron 1 (rs2593813) of the AHSG gene. The G/G genotype for rs2593813 was more common among lean than among obese and overweight individuals (odds ratio = 2.01, P = 0.009), whereas rs2593813 was in strong linkage disequilibrium (|D'| > or = 0.97) with rs4917 and rs4918. Homozygosity for the rs2593813:G-rs4917:Met-rs4918:Ser haplotype conferred an increased risk for leanness (odds ratio=1.90, P = 0.027). rs4917:Met and rs4918:Ser have previously been associated with lower AHSG protein level. A common variant of AHSG, previously associated with a lower AHSG protein level, is thus more common among lean than obese and overweight men, supporting the results from Ahsg knock out mice, namely, that AHSG modulates body mass. PMID- 15806396 TI - Functional promoter polymorphism in the TBX21 gene associated with aspirin induced asthma. AB - Asthma is a phenotypically heterogeneous disorder with many etiologic factors and clinical characteristics. T-bet, a Th1-specific transcription factor of T-box family, has been found to control interferon-gamma (IFN-gamma) expression in T cells. Mice lacking the T-bet gene (tbx21) demonstrate multiple physiological and inflammatory features reminiscent of human asthma. In order to examine whether polymorphisms in the candidate gene, TBX21, located on chromosome 17q21.32, are related to the risk of human asthma phenotypes, we have searched for genetic variations in the human TBX21 gene and identified 24 single nucleotide polymorphisms (SNPs), including five novel SNPs, by direct sequencing in Japanese subjects. Among asthma phenotypes, a promoter -1993T-->C SNP, which is in linkage disequilibrium with a synonymous coding 390A-->G SNP in exon 1, is significantly associated with a risk of aspirin-induced asthma (AIA; P = 0.004, P(c) = 0.016). This association has also been confirmed in additional independent samples of asthma with nasal polyposis (P = 0.008), regardless of aspirin hypersensitivity. Furthermore, our data indicate that the -1993T-->C substitution increases the affinity of a particular nuclear protein to the binding site of TBX21 covering the -1993 position, resulting in increased transcriptional activity of the TBX21 gene. Thus, in addition to the antigen-driven excess Th2 response, increased T bet (and subsequent IFN-gamma) production in human airways of individuals with the -1993T-->C polymorphism could contribute to the development of certain asthma related phenotypes, such as AIA. PMID- 15806397 TI - Ancestral origins of the prion protein gene D178N mutation in the Basque Country. AB - Fatal familial insomnia (FFI) and familial Creutzfeldt-Jakob disease (fCJD) are familial prion diseases with autosomal dominant inheritance of the D178N mutation. FFI has been reported in at least 27 pedigrees around the world. Twelve apparently unrelated FFI and fCJD pedigrees with the characteristic D178N mutation have been reported in the Prion Diseases Registry of the Basque Country since 1993. The high incidence of familial prion diseases in this region may reflect a unique ancestral origin of the chromosome carrying this mutation. In order to investigate this putative founder effect, we developed "happy typing", a new approach to the happy mapping method, which consists of the physical isolation of large haploid genomic DNA fragments and their analysis by the Polymerase Chain Reaction in order to perform haplotypic analysis instead of pedigree analysis. Six novel microsatellite markers, located in a 150-kb genomic segment flanking the PRNP gene were characterized for typing haploid DNA fragments of 285 kb in size. A common haplotype was found in patients from the Basque region, strongly suggesting a founder effect. We propose that "happy typing" constitutes an efficient method for determining disease-associated haplotypes, since the analysis of a single affected individual per pedigree should provide sufficient evidence. PMID- 15806398 TI - Genetic evidence in support of a shared Eurasian-North African dairying origin. AB - The process by which pastoralism and agriculture spread from the Fertile Crescent over the past 10,000 years has been the subject of intense investigation by geneticists, linguists and archaeologists. However, no consensus has been reached as to whether this Neolithic transition is best characterized by a demic diffusion (with a significant genetic input from migrating farmers) or a cultural diffusion (without substantial migration of farmers). Milk consumption and thus lactose tolerance are assumed to have spread with pastoralism and we propose that by looking at the relevant mutations in and around the lactase gene in human populations, we can gain insight into the origin(s) and spread of dairying. We genotyped the putatively causal allele for lactose tolerance (-13910T) and constructed haplotypes from several polymorphisms in and around the lactase gene (LCT) in three North African Berber populations and compared our results with previously published data. We found that the frequency of the -13910T allele predicts the frequency of lactose tolerance in several Eurasian and North African Berber populations but not in most sub-Saharan African populations. Our analyses suggest that contemporary Berber populations possess the genetic signature of a past migration of pastoralists from the Middle East and that they share a dairying origin with Europeans and Asians, but not with sub-Saharan Africans. PMID- 15806399 TI - Molecular characterization of histidinemia: identification of four missense mutations in the histidase gene. AB - Histidinemia (MIM235800) is characterized by elevated histidine in body fluids and decreased urocanic acid in blood and skin and results from histidase (histidine ammonia lyase, EC 4.3.1.3) deficiency. It is the most frequent inborn metabolic error in Japan. Although the original description included mental retardation and speech impairment, neonatal screening programs have identified the majority of histidinemic patients with normal intelligence. Molecular characteristics of histidase in histidinemia have not been determined, and cytogenetically visible deletions of 12q22-24.1 in which histidase gene resides have not been identified in histidinemic patients. In order to investigate whether individuals with this disorder have small deletions, additions, or point mutations in the histidase gene, we screened genomic DNA isolated from 50 histidinemic individuals who were discovered by the neonatal screening program. The methods employed included polymerase chain reaction (PCR) amplification of exons 1-21 of the histidase gene, followed by mutation detection enhancement gel electrophoresis and sequencing of the PCR products displaying heteroduplex bands. Four missense mutations (R322P, P259L, R206T, and R208L), two exonic polymorphisms (T141T c.423A-->T and P259P c.777A-->G), and two intronic polymorphisms (IVS6-5T-->C and IVS9+25A-->G) were identified. The frequencies of each polymorphism estimated either by dot blot allele-specific oligonucleotide hybridization, restriction enzyme digestion, or direct sequencing of the PCR products amplified from 50 unrelated normal individuals were 0.28, 0.30, 0.40, and less than 0.01, respectively. Mutation analysis of one family demonstrated that the patient inherited R322P from the mother and P259L from the father. This report describes the first mutations occurring in the coding region of the histidase structural gene in patients with histidinemia. PMID- 15806400 TI - Calcium-dependent activation of T-lymphocytes. AB - Activation of T-lymphocytes requires stimulation of T-cell receptors (TCR) and co stimulatory signals. Among different signalling cascades, TCR engagement induces Ca(2+) entry through plasma membrane Ca(2+) channels, which is an indispensable step for T-cells to expand clonally and to acquire effector functions. The Ca(2+) channels are activated by depletion of Ca(2+) stores and are called Ca(2+) release-activated Ca(2+) (CRAC) channels. Ca(2+) influx through CRAC channels is also controlled, directly or indirectly, by K(+) channels, Ca(2+)-ATPases, mitochondria, endoplasmic reticulum and Ca(2+) buffers. We review the functional implications of these transporters, organelles and buffers and develop a model of Ca(2+) signal generation that depends mainly on their relative mutual localization. This model offers the possibility of controlling amplitude and kinetics of Ca(2+) signals in T-cells. Decoding of various Ca(2+) signals allows differential activation of the transcription factor families nuclear factor of activated T-cells (NFAT), nuclear factor-kappaB (NF-kappaB) and activator protein 1 (AP-1). Variation of amplitude and kinetics of Ca(2+) signals thus is an important mechanism for modulating the specificity of T-cell responses. PMID- 15806401 TI - Matched adaptations of electrophysiological, physiological, and histological properties of skeletal muscles in response to chronic hypoxia. AB - This study tried to differentiate the consequences of chronic hypoxia on the electrophysiological and physiological properties and the histological characteristics of slow and fast muscles in rats. Animals inhaled a 10% O(2) concentration for a 1-month period. Then, slow [soleus (SOL)] and fast [extensor digitorum longus (EDL)] muscles were analyzed in vitro by physiological and electrophysiological measurements and histological analyses. The results were compared to those obtained in corresponding muscles of an age-matched normoxic group. After exposure to hypoxia: (1) in SOL, there was a tendency to elevated F(max), a significant increase in twitch force and tetanic frequency and a shortening of M-wave duration, and a reduced percentage of type I fibres, whereas the proportion of type IIa fibres doubled; (2) in EDL, F(max) and tetanic frequency were lowered, the muscle became less resistant to fatigue, and the proportion of type IId/x fibres was halved. Then, after 1 month of hypoxia, in the SOL muscle, both the contractile and histological properties resemble those of a fast muscle. By contrast, the EDL became slower, despite its histology was modestly affected. Reduced muscle use in hypoxia could explain the tendency for deteriorating adaptations in EDL, and the faster properties of SOL could result from hypoxia-induced inhibition of the growth-related fast-to-slow shift in muscle fibre types. PMID- 15806402 TI - Endogenous RGS proteins enhance acute desensitization of GABA(B) receptor activated GIRK currents in HEK-293T cells. AB - The coupling of GABA(B) receptors to G-protein-gated inwardly rectifying potassium (GIRK) channels constitutes an important inhibitory pathway in the brain. Here, we examined the mechanism underlying desensitization of agonist evoked currents carried by homomeric GIRK2 channels expressed in HEK-293T cells. The canonical GABA(B) receptor agonist baclofen produced GIRK2 currents that decayed by 57.3+/-1.4% after 60 s of stimulation, and then deactivated rapidly (time constant of 3.90+/-0.21 s) upon removal of agonist. Surface labeling studies revealed that GABA(B) receptors, in contrast to micro opioid receptors (MOR), did not internalize with a sustained stimulation for 10 min, excluding receptor redistribution as the primary mechanism for desensitization. Furthermore, heterologous desensitization was observed between GABA(B) receptors and MOR, implicating downstream proteins, such G-proteins or the GIRK channel. To investigate the G-protein turnover cycle, the non-hydrolyzable GTP analogue (GTPgammaS) was included in the intracellular solution and found to attenuate desensitization to 38.3+/-2.0%. The extent of desensitization was also reduced (45.3+/-1.3%) by coexpressing a mutant form of the Galphaq G-protein subunit that has been designed to sequester endogenous RGS proteins. Finally, reconstitution of GABA(B) receptors with Galphao G-proteins rendered insensitive to RGS resulted in significantly less desensitization (28.5+/-3.2%). Taken together, our results demonstrate that endogenous levels of RGS proteins effectively enhance GABA(B) receptor-dependent desensitization of GIRK currents. PMID- 15806403 TI - [Injury pattern and clinical course of children with multiple injuries in comparison to adults, Ab 11-year analysis at a clinic of maximum utilization]. AB - Due to the small number of polytraumatized children treated in single clinics, only a few studies are available that compare multiply injured children and adults. However, the differences between the two groups with respect to type of injury, injury patterns and pathophysiology may have some crucial effect on the clinical course and are therefore essential for the treatment regime. In this study data about the epidemiology, the preclinical treatment, the clinical course and the complication rates of polytraumatized children and adults were analysed. The aim of this study was to point out the main differences between the two collectives and to compare the results with the current literature. Traffic accidents were the leading cause for trauma (children 77.8%, adults 62.6%). Children more often were transported by helicopter than adults. In both age groups head injuries and fractures of the extremities were most frequent. Adults had a prolonged ventilation time as well as an increased stay in ICU and a regular ward. PMID- 15806404 TI - Squalene aspiration pneumonia in children: radiographic and CT findings as the first clue to diagnosis. AB - BACKGROUND: The diagnosis of squalene aspiration pneumonia in children is often difficult because of minimal non-specific symptoms. OBJECTIVE: To investigate the radiological findings of squalene aspiration pneumonia in children. MATERIALS AND METHODS: We reviewed the chest radiographs (n = 8) and CT scans (n = 7), including high-resolution CT (n = 3), of eight patients (four boys, four girls; age 3 months to 6 years) with squalene aspiration pneumonia. All patients presented minimal symptoms. RESULTS: Chest radiographs showed right-sided predominantly parahilar infiltrations. The extent and the opacity of the lesions decreased slowly during the follow-up period (mean 5.4 months) after halting the exposure. On CT, affected areas appeared as dense consolidations surrounded by ground-glass opacities showing a crazy-paving pattern in a geographic lobular distribution in all patients. The lesions were predominantly in the right lung and dependent areas in all patients and extensively involved all pulmonary lobes in five patients. CONCLUSIONS: These radiological findings, although non specific, can lead to an appropriate diagnosis, particularly when patients present few symptoms. PMID- 15806405 TI - Costal osteochondroma presenting as haemothorax and diaphragmatic laceration. AB - We report the unusual case of an 11-year-old girl who suffered a large haemothorax and lacerated left hemidiaphragm because of a costal osteochondroma arising from the left sixth rib near the costochondral junction. Surgical excision of the osteochondroma was performed and the lacerated diaphragm was repaired. PMID- 15806409 TI - [Implant infections. What must an internist know?]. AB - Implant-associated infections are a major challenge for the primary care physician because the chance of cure depends on the time interval between infection and start of adequate surgical and antimicrobial treatment. On the other hand, rapid therapy should not prevent from correct microbiological sampling. Since long-term antibiotic therapy is required, the microorganism and its susceptibility pattern should be known. Thus, the primary care physician should know signs and symptoms of these rare infections and rapidly involve the specialist. The main task of the specialist is to confirm the diagnosis and to decide whether the implant has to be removed, or whether debridement with retention is a feasible option to eradicate infection. In this review, infections associated to vascular grafts, to pacemakers and to arthroplasties are presented. PMID- 15806410 TI - [Diagnosis and therapy of thyroid nodules]. AB - Thyroid nodules and goitre can be diagnosed in up to 50% in populations living in iodine deficiency areas. Because of the necessity to exclude malignancy they therefore represent a significant diagnostic and economic problem. Sonography as well as TSH determination are the basic constituents of any thyroid diagnostic work up. Thyroid scintigraphy should be performed with any solitary thyroid nodule >10 mm if the scintigraphic result (together with the sonographic result) is likely to influence the treatment. Except of hot nodules any thyroid nodule should be evaluated by fine needle aspiration biopsy. Because of the lack of controlled studies including sufficient numbers of patients, there is a lack of evidence for some aspects of our everyday clinical practice. The aim of this article is therefore to summarize latest results on pathogenesis, diagnostic tools and recommendations concerning therapy and follow up. PMID- 15806411 TI - [Cardiovascular end organ impairment due to hypertension]. AB - Arterial hypertension is the most frequent cause of pressure overload on the left ventricle. Longer lasting arterial hypertension leads to hypertension-specific organ manifestations summarized as "hypertensive heart disease". Hypertensive heart disease comprise the manifestation of stenosis in epicardial arteries, hypertensive microvascular disease, ischemic cardiomyopathy, left ventricular hypertrophy, endothelial dysfunction, increased sympathetic drive and degeneration of aortic valve. Diastolic dysfunction and reduced coronary flow reserve can be evaluated as early markers of hypertensive heart disease. These alterations lead to the major clinical manifestations of hypertensive heart disease that are symptoms of reduced coronary insufficiency with typical angina pectoris, but also of symptoms of heart failure (systolic and diastolic dysfunction) and arrhythmia. Different non-invasive and invasive procedures are available for screening and follow-up of patients with hypertensive heart disease. Primary therapeutic target is, apart from lowering blood pressure, to reverse cardiac manifestations of arterial hypertension using specific therapeutic algorithms. PMID- 15806412 TI - [Cardiogenic shock in a 61 year old female with recurrent panic attacks]. AB - A 61 year old patient in a severely reduced general condition complaining of epigastric pain was admitted to our emergency room. Because of elevated troponin T and creatine kinase levels and severely reduced left ventricular function as seen in echocardiography as well as negative T-waves in the anterolateral ECG leads we suspected an acute coronary syndrome. The patient underwent coronary angiography after intubation and a coronary artery disease was excluded. The initial therapy included the use of an intraaortic balloon pump, volume and catecholamine administration controlled by a pulmonary artery catheter. Conducting an abdominal ultrasound we detected a mass at the right adrenal gland. The suspected diagnosis of pheochromocytoma was confirmed by elevated catecholamine levels in the urine and a CT scan. After recompensation and subsequent administration of phenoxybenzamine a benign pheochromocytoma was resected. A massive catecholamine secretion due to a pheochromocytoma can lead to a cardiogenic shock and multiple organ failure. In patients with recurrent panic attacks and hypertension a pheochromocytoma should be included in the differential diagnosis. PMID- 15806413 TI - [Pharmacologic therapy for inflammatory bowel diseases: hopes, disappointments]. AB - Because the etiology of inflammatory bowel diseases is unclear, no causative therapy is available. However, pathophysiology of the disease offers a lot of possibilities to disrupt the inflammatory cascade that maintains the inflammatory process. The aim of every therapy is to maintain remission as long as possible and to amend the natural course of the disease. Pharmacotherapy includes 5 Aminosalicylates, glucocorticoids, immunosupressants (methotrexate, azathioprine) as well as specific pharmacologic interventions like monoclonal antibodies directed against TNF-alpha (Infliximab). Important supportive tools are available to improve symptoms like diarrhea and pain. Dietetic treatment and surgical procedures represent important alternatives or supplement pharmacotherapeutic interventions. PMID- 15806414 TI - [Renovascular hypertension--diagnosis and therapy]. AB - Atherosclerotic renal artery stenosis accounts for most cases of renovascular hypertension (RVH). Hypertensive patients with clinical features suggesting RVH should be submitted to further noninvasive evaluation including duplex Doppler ultrasonography, CT- or MR angiography. Invasive evaluation by contrast-enhanced angiography confirms the diagnosis of renal artery stenosis. However, neither diagnostic test reliably predicts the course of hypertension after revascularisation. The therapeutic approach in hypertensive patients with hemodynamically important renal artery stenosis includes medical or invasive therapy (renal percutaneous transluminal angioplasty, PTRA; renal arterial stent placement, PTRAS; surgical revascularisation). Randomized trials comparing invasive and conservative approaches demonstrated no differences in blood pressure control or renal function. Only patients with clear clinical indications should be submitted to interventional procedures as PTRA, PTRAS and surgical vascular intervention. PMID- 15806415 TI - In vivo antitumor activity by 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone in a solid human carcinoma xenograft model. AB - Previously we have shown that 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone (DMC), which is isolated from the buds of Cleistocalyx operculatus, significantly inhibits the growth of human liver cancer SMMC-7721 cells and is able to induce apoptosis of SMMC-7721 cells in vitro. Here we report the antitumor effects of DMC in vivo, using a solid human tumor xenograft mouse model using human liver cancer SMMC-7721 cells. The average tumor weights in the control group and in mice injected with 150 mg/kg DMC were 1.42+/-0.11 g and 0.59+/-0.12 g, respectively. Flow cytometric analysis of the tumor cell population demonstrated an aneuploid peak (representing 33.60+/-0.80% of the total in mice injected with 150 mg/kg DMC). To our knowledge, this is the first time that chalcone compounds have been applied to a human tumor xenograft model. PMID- 15806416 TI - Randomized clinical trial to compare the effects of methadone and buprenorphine on the immune system in drug abusers. AB - RATIONALE: Buprenorphine may be a useful alternative option to methadone in addicts. Opioids can produce severe changes in the immune system. OBJECTIVES: The objectives of this study are to compare the effect of sublingual buprenorphine and methadone on the immune system and to compare the two substances on the drying-out program compliance. METHODS: We studied 62 randomized outpatients for a period of 12 months. Subjects (55 males and 7 females; mean age 25+/-4 years; average history of heroin abuse being 2 years) on maintenance treatment were assigned in two groups (A and B). Methadone chloride (medium dose 100 mg/day) was administered to group A, whereas group B received sublingual buprenorphine (32.40+/-2.8 mg/day). Urine toxicological screening, plasma levels of TNF-alpha interleukin-1, interleukin-beta, lymphocyte CD14 and a self-rating depression questionnaire were measured. RESULTS: Urine screening was negative for opiates in 17.6% of group A and in 10.7% of group B (p<0.001; r = 0.62). Depression score was 62+/-2 in group A and 55+/-3 in group B (p < 0.01). Cytokine and CD14 revealed higher concentrations both in groups A and B without significant differences (p > 0.05) between the two groups. CONCLUSIONS: The effects of buprenorphine and methadone tested on the immune system were overlapping in our patients. The elevated cytokine levels observed may suggest that the two drugs stimulate immunologic hyperactivation of an immune system that was formerly inhibited by heroin. Furthermore, our data suggest that buprenorphine can be a valid alternative to methadone in maintenance treatment of chronic heroin abuse and referred a marked decline in depression. PMID- 15806417 TI - Combined treatment of ascorbic acid or alpha-tocopherol with dopamine receptor antagonist or nitric oxide synthase inhibitor potentiates cataleptic effect in mice. AB - RATIONALE: Drugs like haloperidol (Hal) that decrease dopamine (DA) neurotransmission in the striatum induce catalepsy in rodents and Parkinson disease-like symptoms in humans. Nitric oxide synthase (NOS) inhibitors interfere with motor activity, disrupting rodent exploratory behavior and inducing catalepsy. Catalepsy induced by NOS inhibitors probably involves striatal DA mediated neurotransmission. Antioxidants such as ascorbic acid (vitamin C) and alpha-tocopherol (vitamin E) have also been shown to interfere with movement modulation and the DA system. OBJECTIVE: The objective of the study is to investigate if the antioxidants vitamins C and E would influence the catalepsy produced by Hal and NOS inhibitors. METHODS: The effects of the following treatments on catalepsy were examined using the hanging-bar test on male Swiss mice (25-30 g): (1) vitamin C (30-1,000 mg/kg)xHal (1 mg/kg); (2) vitamin C (90 1,000 mg/kg)xN (G)-nitro-L: -arginine (LNOARG, 10 and 40 mg/kg); (3) vitamin C (300 mg/kg)xN (G)-nitro-L: -arginine methylester (LNAME, 20-80 mg/kg); (4) vitamin C (300 mg/kg) x 7-nitroindazole (7NI, 3-50 mg/kg); (5) vitamin C (90 mg/kg i.p.) x LNOARG [40 mg/kg twice a day during 4 days (subchronic treatment)]; (7) vitamin E (3-100 mg/kg) x Hal (1 mg/kg); and (6) vitamin E (3-100 mg/kg) x LNOARG (40 mg/kg). RESULTS: Vitamin C enhanced the catalepsy produced by NOS inhibitors and Hal. Treatment with vitamin C did not affect tolerance to LNOARG cataleptic effect induced by subchronic treatment. Vitamin E potentiated the catalepsy induced by LNOARG at all doses tested; in contrast, catalepsy induced by Hal was enhanced only by the dose of 100 mg/kg. CONCLUSIONS: Results support an involvement of dopaminergic and nitrergic systems in motor behavior control and provide compelling evidence that combined administration of the antioxidants vitamins C and E with either Hal or NOS inhibitors exacerbates extrapyramidal effects. Further studies are needed to assess possible clinical implications of these findings. PMID- 15806418 TI - [Apraxias]. AB - Apraxias are deficits in higher motor behaviour that are not primarily caused by elementary deficits of the sensorimotor system, communication problems, or dementia. These patients present with deficits such as imitating meaningful or meaningless gestures and in dexterity or purposeful use of objects. The different forms of apraxia originate from lesions of different levels/structures of the motor system, reflecting its complexity. Apraxias are caused by deficits in motor programmes generated in the frontal motor areas, in modality-specific higher sensorimotor control, or at the highest level of motor planning and motor conception. The types of apraxia differentially affect activities of daily living and hence show marked differences in the prognosis of recovery and the physiotherapeutic treatment required. Therefore, appropriate diagnosis and treatment of the different forms are of foremost clinical importance. PMID- 15806419 TI - [MS registry in Germany--design and first results of the pilot phase]. AB - In the summer of 2001, a nationwide epidemiological multiple sclerosis (MS) register was initiated under the auspices of the German MS Society (DMSG). This project aimed at collecting epidemiological data on the number of patients with MS, course of the disease, and their social situation in Germany. During the 2 year pilot phase, five MS centers with various regional differences and treatment methods participated, leading to a representative selection of patients. In December 2003, standardised data sets of 3,458 MS patients were available for evaluation. After examining the quality of the data, 3,223 sets remained for further analysis. The demographics were similar to those obtained from other epidemiological studies: 72% of the patients were female, mean age was 42.9+/ 11.2 years, mean disease duration 12.6+/-8.7 years, and 64% suffered from the relapsing-remitting form of the disease. The median EDSS was 3.0, and 69% of patients had an EDSS 1.4 mg/dL [>124 micromol/L]; women, >1.2 mg/dL [>106 micromol/L]; odds ratio, 3.21; 95% confidence interval, 1.32 to 7.76 and odds ratio, 5.39; 95% confidence interval, 2.08 to 13.99, respectively). CONCLUSION: This is the first study to show an association of periodontal disease with prevalent renal insufficiency. A prospective study is necessary to determine the exact nature of the observed relationship. PMID- 15806468 TI - Quality of life in chronic kidney disease (CKD): a cross-sectional analysis in the Renal Research Institute-CKD study. AB - BACKGROUND: Health-related quality of life (QOL) is an important measure of how disease affects patients' lives. Dialysis patients have decreased QOL relative to healthy controls. Little is known about QOL in patients with chronic kidney disease (CKD) before renal replacement therapy. METHODS: The Medical Outcomes Study Short Form-36 (SF-36), a standard QOL instrument, was used to evaluate 634 patients (mean glomerular filtration rate [GFR], 23.6 +/- 9.6 mL/min/1.73 m2 [0.39 +/- 0.16 mL/s/1.73 m2]) enrolled in a 4-center, prospective, observational study of CKD. SF-36 scores in these patients were compared with those in a prevalent cohort of hemodialysis (HD) patients and healthy controls (both from historical data). QOL data also were analyzed for correlations with GFR and albumin and hemoglobin levels in multivariable analyses. RESULTS: Patients with CKD had higher SF-36 scores than a large cohort of HD patients (P < 0.0001 for 8 scales and 2 summary scales), but lower scores than those reported for the US adult population (P < 0.0001 for 7 of 8 scales and 1 of 2 summary scales). Patients with CKD stage 4 had lower QOL scores than patients with CKD stage 5, although differences were not significant. Hemoglobin level was associated positively with higher mental and physical QOL scores (P < 0.05) in all individual and component scales except Pain. CONCLUSION: SF-36 scores were higher in this CKD cohort compared with HD patients, but lower than in healthy controls. GFR was not significantly associated with QOL. Hemoglobin level predicted both physical and mental domains of the SF-36. Longitudinal studies are needed to define at-risk periods for decreases in QOL during progression of CKD. PMID- 15806469 TI - Microscopic hematuria in pregnancy: relevance to pregnancy outcome. AB - BACKGROUND: The significance of dipstick or microscopic hematuria in pregnancy is uncertain, with some studies suggesting this is associated with a greater risk for preeclampsia. We sought to determine the prevalence and clinical significance of microscopic hematuria during pregnancy. METHODS: This was a prospective case control study in the antenatal Clinic of St George Hospital, Kogarah, Australia, a teaching hospital without tertiary referral antenatal care, with approximately 2,600 deliveries per year. One thousand pregnant women attending for routine antenatal care were invited to have a routine urinalysis performed and be referred to a nephrology clinic for further investigation if dipstick microscopic hematuria was detected on more than 1 occasion before 32 weeks' gestation. Main outcome measures were the prevalence of dipstick hematuria, prevalence of hematuria confirmed by urine microscopy, and the development of preeclampsia or gestational hypertension or delivery of a small-for-gestational-age baby. RESULTS: One hundred seventy-eight of 902 women (20%) who entered the study had dipstick hematuria on at least 2 occasions in pregnancy; 66 of 126 women (53%) who had hematuria before 32 weeks attended the nephrology clinic, where microscopic hematuria was confirmed in 40 women (61%). Renal imaging results were normal in all except 1 woman, and all women had a serum creatinine level of 0.90 mg/dL or less (< or =80 micromol/L). The development of preeclampsia or gestational hypertension or delivery of a small-for-gestational-age baby were similar in women with and without dipstick hematuria. Microscopic hematuria persisted in half (15 women) of those who attended for follow-up after 3 months postpartum. CONCLUSION: Dipstick hematuria is very common during pregnancy, but rarely signifies a disorder likely to impact on the pregnancy outcome. Postpartum follow-up is recommended to detect women who have persistent hematuria and presumed underlying mild glomerulonephritis. PMID- 15806470 TI - Immunoperoxidase versus immunofluorescence in the assessment of human renal biopsies. AB - BACKGROUND: For half a century, immunofluorescence (IF) on frozen sections has been the gold standard for immunohistochemical evaluation of renal biopsy specimens. In routine diagnostic immunohistopathologic evaluation, traditional IF has been replaced to a large extent by immunoperoxidase (IP) methods applied to paraffin sections of formaldehyde-fixed tissue. This is caused in part by the practical disadvantages inherent in the IF method, eg, separate tissue specimen and handling, UV microscopy, fading and impermanence of the label-making archiving, and difficult later investigation. Our aim for the present study is to evaluate IP as an alternative to IF in the diagnostic assessment of renal biopsy specimens. METHODS: Proteolytic antigen retrieval, antibodies effective on deparaffinized sections, a sensitive detection system (Dako EnVision HRP; Dako, Copenhagen, Denmark), and a standardized and rigorously controlled procedure were applied to a series of renal biopsy specimens (n = 81) previously classified by means of light microscopy (LM) and IF. Staining for immunoglobulin G (IgG), IgA, IgM, C1q, and C3c were recorded as positive or negative for IF and IP in paired proportions, presuming that IF was the test standard. RESULTS: Concordant observations were 71% for all (282 of 398 observations), 82% for IgG (65 of 79 observations), and 89% for IgA (72 of 81 observations). The majority of discordant observations (74 of 116 observations) were positive by means of IP, with mesangial deposits of IgM and C1q that were not found by IF. Statistically, there was no significant difference in outcomes between IF and IP for IgG, IgA, and C3c ( P > 0.2). In addition, IP staining allowed simultaneous evaluation of tissue by LM and therefore correlation between tissue structure and immune deposits not readily attained by IF. CONCLUSION: In the present study, it is documented that for the detection of IgG, IgA, and C3c, IP applied to protease digested deparaffinized sections of formaldehyde-fixed renal tissue is, with few exceptions, equal to IF on frozen sections. The EnVision HRP method used here is several times more effective in terms of primary antibody dilution than earlier existing IP methods, and because the avidin-biotin system is not involved, very little nonspecific background staining will occur. Discordant observations (116 of 398 observations; 29%) were in the majority (91 of 116 observations) due to positive IP findings of IgM and C1q, which deserve additional investigation. PMID- 15806471 TI - Hepatitis C infection is acquired pre-ESRD. AB - BACKGROUND: Hepatitis C is more prevalent in patients with end-stage renal disease (ESRD) than in the general population. Previously, infection from nosocomial sources was an important cause for this increase. In the past decade, the risk from these sources has lessened, but the estimated percentage of patients with ESRD with hepatitis C virus (HCV) antibody (anti-HCV) has not changed. METHODS: This is a prospective observational study of hemodialysis patients in 4 urban units in Birmingham, AL. Testing for anti-HCV was performed at the initiation of dialysis therapy and then yearly from August 1998 through August 2004. Race, sex, age, ESRD date, comorbid conditions, and outcomes were recorded. RESULTS: The prevalence of anti-HCV was 16.8% in 860 patients (89% black). Patients new to ESRD had a prevalence of 14.4%. Seroconversion in previously anti-HCV-negative patients was 2.5%. Anti-HCV occurred in black men 3 times more than in black women (odds ratio, 3.3; 95% confidence interval, 2.2 to 5.0). Mean age at ESRD was significantly younger in anti-HCV-positive than HCV negative patients (47.3 versus 54.1 years; P < 0.0001). Age, race, sex, and history of drug abuse were predictors of anti-HCV positivity. CONCLUSION: The prevalence of anti-HCV among patients with ESRD varies from community to community. New patients are major contributors to the prevalence of anti-HCV in patients with ESRD, particularly those who are younger, male, black, or have a history of drug use. Measuring anti-HCV levels in patients with chronic kidney disease may help identify those at risk for additional disorders. PMID- 15806472 TI - Association of physical activity with mortality in the US dialysis population. AB - BACKGROUND: It is unclear whether the protective benefits of regular physical activity on mortality extend to patients with end-stage renal disease (ESRD). We tested this hypothesis in a national cohort of new patients with ESRD in the United States. METHODS: Data for a subset of patients (n = 2,507; 62%) from the Dialysis Morbidity and Mortality Wave 2 study were used to explore the associations of exercise and limitations in physical activity with mortality. RESULTS: Overall, 56% of patients exercised less than once a week, whereas the remainder reported more frequent physical activity; 2 to 3 times/wk in 18%, 4 to 5 times/wk in 6%, and daily exercise in 20%. Severe limitations in vigorous and moderate physical activities were reported by 75% and 42%, respectively. Mortality risks were greatest for those with severe limitations in either moderate (relative risk [RR], 1.72; 95% confidence interval [CI], 1.44 to 2.05) or vigorous physical activities (RR, 1.51; 95% CI, 1.20 to 1.90) compared with those reporting minimal or no limitations. Conversely, mortality risks were lower for patients who exercised 2 to 3 (RR, 0.74; 95% CI, 0.58 to 0.95) or 4 to 5 times/wk (RR, 0.70; 95% CI, 0.47 to 1.07), whereas no advantage was associated with daily exercise (RR, 1.06; 95% CI, 0.86 to 1.30). CONCLUSION: Although limitations in physical activity are common among new patients with ESRD in the United States and correlate highly with increased mortality risk, this study shows an association of frequent exercise of up to 4 to 5 times/wk with improved survival. The surprising lack of association of daily exercise with increased survival deserves additional study. PMID- 15806473 TI - Hyperhomocysteinemia and arteriovenous fistula thrombosis in hemodialysis patients. AB - BACKGROUND: To date, the relationship between vascular access (VA) failure and plasma total homocysteine level has been investigated only in mixed dialysis populations (ie, patients with a native arteriovenous [AV] fistula or arterial graft), whereas almost no data exist for hemodialysis patients with a native AV fistula. METHODS: In this prospective cohort study, we examined the relationship between plasma total homocysteine level and the methylenetetrahydrofolate reductase (MTHFR) gene and VA-related incident morbidity in a cohort of 205 hemodialysis patients, all with a native AV fistula. RESULTS: During follow-up, 78 patients experienced 1 or more VA thrombotic episodes. Patients with incident VA thrombosis had a significantly greater plasma total homocysteine level compared with patients without this event (P = 0.046). In Kaplan-Meier survival analysis, the hazard ratio for VA thrombosis increased in parallel with homocysteine level, such that patients in the third homocysteine level tertile had a relative risk for this outcome 1.72 times (95% CI, 1.21 to 2.24) greater than in those in the first tertile (log-rank test, 6.81; P = 0.009). In a multiple Cox regression model, plasma total homocysteine level was confirmed to be an independent predictor of AV fistula outcome. Plasma total homocysteine level was significantly greater (P < 0.001) in patients with the TT genotype of the MTHFR gene than in those with the CT or CC genotype. CONCLUSION: VA thrombosis in dialysis patients is associated with hyperhomocysteinemia. Intervention studies are needed to clarify whether decreasing plasma homocysteine concentrations may prevent VA failure in hemodialysis patients. PMID- 15806474 TI - Vascular access survival in children and young adults receiving long-term hemodialysis. AB - BACKGROUND: The delivery of long-term hemodialysis therapy in children is complicated by smaller vascular caliber and the potential lifelong requirement for hemodialysis access. Various factors have resulted in the increased use of cuffed central venous catheters (CVLs) in preference to autologous arteriovenous fistulae (AVFs) and arteriovenous synthetic grafts (AVGs). The aim of this study is to compare CVL, AVF, and AVG survival and determine factors affecting their survival. METHODS: A 20-year retrospective study was undertaken of pediatric patients receiving long-term hemodialysis therapy. Age, height, weight, body mass index, and sex were noted at each procedure, in addition to the presence of hypoalbuminemia, underlying diagnosis, type and site of vascular access, and effect of previous access surgery. The grade of operator also was noted. RESULTS: Three hundred four vascular access procedures were performed on 114 patients, with a median age at initial access formation of 12.0 years (range, 4 weeks to 21.9 years). The most common procedure was CVL insertion (182 procedures) and then AVF formation (107 procedures), with only 15 AVGs created. Median censored survival was 3.14 years (95% confidence interval, 1.22 to 5.06) for AVFs and 0.6 years (95% confidence interval, 0.20 to 1.00) for CVLs. Factors adversely affecting vascular access survival were younger age, trainee operator, presence of hypoalbuminemia, and type of access undertaken, with AVF better than CVL. CONCLUSION: This study shows increased survival of AVFs over CVLs and AVGs. Vascular access in children and adolescents may impact on future dialysis accessibility and should be undertaken by those most experienced in each technique. PMID- 15806475 TI - Circulating soluble vascular cell adhesion molecule 1: relationships with residual renal function, cardiac hypertrophy, and outcome of peritoneal dialysis patients. AB - BACKGROUND: Vascular cell adhesion molecule 1 (VCAM-1) is involved in leukocyte endothelial cell interaction and has a pivotal role in inflammation. Whether it contributes to excessive mortality in dialysis patients remains uncertain. In this study, we examined circulating soluble VCAM-1 (sVCAM-1) in relation to different clinical and biochemical parameters, as well as mortality and cardiovascular events, in peritoneal dialysis (PD) patients. METHODS: Values for serum sVCAM-1, together with C-reactive protein (CRP), homocysteine, albumin, lipid profile, blood hemoglobin, and indices of dialysis adequacy, were determined at study baseline, and echocardiography was performed in 160 long-term PD patients. Patients were followed up for a mean of 35 +/- 16 (SD) months. RESULTS: Serum sVCAM-1 levels were elevated in our continuous ambulatory PD (CAPD) patients and showed a negative correlation with residual glomerular filtration rate (GFR; P < 0.001) and low-density lipoprotein (LDL) cholesterol level (P = 0.004), but a positive correlation with left ventricular mass index (P = 0.025). Using Kaplan-Meier analysis, overall survival rates at 2 years were 96.2%, 75.2%, and 50.6% for patients in the lower, middle, and upper tertiles of sVCAM-1 levels, respectively (P < 0.0001). Fatal and nonfatal cardiovascular event-free survival rates were 58.2%, 56.9%, and 19.4% for patients in the lower, middle, and upper tertiles, respectively (P < 0.0001). Using Cox regression analysis with adjustment for confounding covariates, every 100-ng/mL increase in sVCAM-1 level was associated with 8% (95% confidence interval, 1.03 to 1.13) and 5% (95% confidence interval, 1.00 to 1.10) increases in risk for death and fatal and nonfatal cardiovascular events, respectively. Its significance for all-cause mortality remained with additional adjusting for LDL cholesterol level, but was lost when adjusting for residual GFR. Its association with cardiovascular events became insignificant when adjusting for LDL cholesterol level or residual GFR. Furthermore, patients with both sVCAM-1 and CRP levels elevated at the 50th percentile or greater were associated with the greatest death and fatal and nonfatal cardiovascular event rates compared with those with either CRP or sVCAM 1 level elevated at the 50th percentile or greater. CONCLUSION: Circulating sVCAM 1 levels show an important link with residual renal function, LDL cholesterol level, and cardiac hypertrophy in CAPD patients. Furthermore, residual renal function, which correlates inversely with circulating sVCAM-1 level, shows an important association with all-cause mortality and cardiovascular events and displaces sVCAM-1 level from the models for all-cause mortality and future cardiovascular events in CAPD patients. Additional study is needed to explore possible mechanistic links between inflammation, soluble adhesion molecules, residual renal function, and cardiac hypertrophy in CAPD patients. PMID- 15806476 TI - Association of patient autonomy with increased transplantation and survival among new dialysis patients in the United States. AB - BACKGROUND: It is unclear whether patients with chronic kidney disease who are more autonomous in medical decision making have better outcomes than those who are not. We examined the contribution of patient autonomy to treatment selection (peritoneal dialysis versus hemodialysis) and subsequent association with transplantation and survival. METHODS: Data were obtained from the Dialysis Morbidity and Mortality Study Wave 2, a national random sample of 4,025 new dialysis patients enrolled during 1996 and 1997 and followed up until October 31, 2001. Responders were asked to quantify their contribution to treatment selection and were grouped based on perceived degree of participation as patient led, team led, or patient and team led. Groups were compared and subsequent outcomes were evaluated by using Cox regression. RESULTS: Six hundred thirty-six patients (26.3%) reported a patient-led decision, 860 patients (35.6%) reported a team-led decision, and 922 patients (38.1%) reported a patient-and-team-led decision in treatment assignment. Unadjusted death rates were significantly lower (127 versus 159 versus 207 deaths/1,000 patient-years at risk; P < 0.0001), and transplantation rates were significantly higher (103 versus 88 versus 41 transplantations/1,000 patient-years at risk; P < 0.0001) for patients reporting the greatest contribution to modality selection. With adjustment for case mix, mortality risks were lowest (relative risk [RR], 0.84; 95% confidence interval [CI], 0.71 to 0.99) and transplantation rates were highest (RR, 1.44; 95% CI, 1.07 to 1.93) for the patient-led group. CONCLUSION: Although the contribution of patient selection factors cannot be completely ignored, this analysis supports an association of patient autonomy with transplantation and survival. Greater efforts to empower patients with chronic kidney disease during the period before end-stage renal disease may improve clinical outcomes. PMID- 15806477 TI - An unusual case of pulmonary-renal syndrome associated with defects in type IV collagen composition and anti-glomerular basement membrane autoantibodies. AB - Commercial serological assays for the presence of anti-glomerular basement membrane (GBM) antibodies are thought to be indicative of Goodpasture's syndrome. We report a case in which commercial tests inaccurately suggested that a patient with a pulmonary-renal syndrome had Goodpasture's disease. Additional laboratory testing using recombinant type IV collagen NC1 domain proteins showed that the autoantibodies in question were not directed against the Goodpasture antigen (the alpha3NC1 domain), but against the alpha2NC1 domain of type IV collagen. Our findings represent the first known case of human autoantibodies to the alpha2NC1 domain. Further investigation showed that this patient has decreased alpha3 and alpha5 chain expression in the GBM and defects in type IV collagen, resembling abnormalities in patients with Alport's syndrome. PMID- 15806478 TI - Fanconi's syndrome induced by a monoclonal Vkappa3 light chain in Waldenstrom's macroglobulinemia. AB - Fanconi's syndrome (FS) is a disorder of sodium-dependent proximal tubule reabsorption, which may complicate plasma cell disorders producing a free monoclonal light chain (LC). FS often occurs in the setting of smoldering myeloma and features cytoplasmic crystalline inclusions of monoclonal kappa LC in proximal tubular cells and malignant plasma cells. Although the clinical and pathological presentation may vary, including lack of crystal formation, monoclonal kappa LCs that underlie FS show a striking genetic and biochemical homogeneity: they almost always belong to the Vkappa1 subgroup of variability and originate from 2 germline genes, O2/O12 or O8/O18. Their variable domain sequences present unusual hydrophobic residues, responsible for the resistance to proteolysis, which leads to LC accumulation in the endocytic compartment of proximal tubule cells. We report a patient with slowly progressive Waldenstrom's macroglobulinemia and full-blown FS with accumulation of a monoclonal kappa LC within proximal tubules, but no detectable crystalline organization. This LC, which belonged to the unusual Vkappa3 subgroup and derived from the L2/L16 germline gene, showed no common substitution with previously described FS kappaI LC and was sensitive to trypsin digestion. These data show that molecular and biochemical characteristics of kappa LCs in patients with FS are more heterogeneous than initially suspected. Mechanisms other than resistance of LCs to endosomal proteolysis probably are involved in the pathogenesis of FS associated plasma cell dyscrasias. PMID- 15806479 TI - Pulmonary-renal syndrome in a newborn with placental transmission of ANCAs. AB - Despite clinical and experimental data suggesting a direct relationship between antineutrophil cytoplasmic antibodies (ANCAs) and disease activity in patients with microscopic polyangiitis (MPA), the causal relationship between perinuclear ANCAs specific for myeloperoxidase (MPO-ANCA) and disease manifestations has been controversial. We describe the case of a woman with a history of pulmonary-renal syndrome caused by MPA whose disease became clinically and serologically active during pregnancy. Forty-eight hours after delivery, the newborn developed pulmonary hemorrhage and abnormalities in renal function. The newborn's cord blood showed an immunoglobulin G MPO-ANCA level identical to that of the mother's serum, indicating passive transfer of the antibody to the neonate. Our findings represent the first human model supporting the interpretation that MPO-ANCAs were immunopathogenic. PMID- 15806480 TI - Thrombotic thrombocytopenic purpura in a patient treated with imatinib mesylate: true association or mere coincidence? AB - Thrombotic thrombocytopenic purpura (TTP), characterized by thrombocytopenia and microangiopathic hemolytic anemia, is a relatively rare disorder. The majority of cases have no defined causes. TTP has been reported in association with many drugs, but not with imatinib mesylate. We report a 22-year-old African-American woman who developed idiopathic hypereosinophilic syndrome. She was treated with imatinib mesylate and subsequently developed microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure. TTP was diagnosed. A kidney biopsy was performed and was diagnostic of thrombotic microangiopathy. The patient was treated with plasma exchange and hemodialysis. Her eosinophilia resolved, but she remained dialysis dependent. To our knowledge, this is the first case report of the possible association between imatinib mesylate and TTP. PMID- 15806481 TI - Needed: tailored exercise regimens for kidney transplant recipients. PMID- 15806483 TI - Nephrotic syndrome in patients with peripheral blood stem cell transplant. PMID- 15806485 TI - Efficacy of furosemide in ARF. PMID- 15806486 TI - Plasma fibrinogen and dialysis. PMID- 15806487 TI - Coronary artery calcification and impaired renal function. PMID- 15806488 TI - Cyclophosphamide or chlorambucil therapy is indicated in idiopathic membranous nephropathy with strong risk factors for progression. PMID- 15806502 TI - K/DOQI clinical practice guidelines for cardiovascular disease in dialysis patients. PMID- 15806503 TI - Where is your manuscript? PMID- 15806504 TI - Nursing students' perceptions of a career in nursing and impact of a national campaign designed to attract people into the nursing profession. AB - The study assessed perceptions about a career in nursing, the nursing shortage, decision to enroll in a nursing education program, and awareness and effect of the Johnson & Johnson Campaign for Nursing's Future. Data were obtained from a survey administered to a national sample of 496 nursing students. Although most students believe nursing is physically challenging and there is inadequate respect and recognition of nurses, most agree nursing is a good career for men and for people who have academic ability and want a secure job. Although the majority believes the nursing shortage will increase stress on nurses, lower the quality of care, and fail to improve working conditions, most students also expect the shortage will lead to higher pay and more job choices. Information and advice from practicing nurses was the most influential factor in deciding to become a nurse, and friends, parents, and other family members also had a positive influence. Two thirds are aware of the campaign, almost all said it made them feel good about becoming a nurse, and students who are White and enrolled in baccalaureate programs were more likely to be aware of the campaign. Future recruitment initiatives should strengthen the focus on men and minorities, parents and friends, and practicing nurses and nursing educators. PMID- 15806505 TI - Development of case statements in academic administration: a proactive method for achieving outcomes. AB - The complex nature of higher education presents academic administrators with unique challenges to communicate vision and strategic direction to a variety of internal and external audiences. The administrator must be prepared to engage in persuasive communication to describe the needs and desired outcomes of the academic unit. This article focuses on the use of the case statement as a communication tool for the nursing academic administrator. The case statement is a form of persuasive communication where a situation or need is presented in the context of the mission, vision, and strategic direction of a group or organization. The aim of the case statement is to enlist support in meeting the identified need. Fundamental assumptions about communicating case statements are described, as well as guidelines for how the academic administrator can prepare themselves for using the case statement method. PMID- 15806506 TI - Practice what you teach: a case study of ethical conduct in the academic setting. AB - Nursing faculty teach ethics and ethical behavior in undergraduate and graduate curriculum. In this article, a case study is presented that illustrates a breach of ethical behavior and conduct in the academic setting by both faculty and students. The decision-making process used to resolve this dilemma by the chair, the associate dean, and a faculty member relied on a dialectic approach that looked at philosophical underpinnings, historical background of nursing ethics, and university- and schoolwide policies and procedures. The conversations facilitated the ethical resolution to the dilemma raised in the case study as well as the recognition of additional issues for consideration. The authors uncovered compelling questions that included, "What is meant by ethical conduct in the classroom?," "How do we teach it?," and "How do we practice it?" The purpose of this article is to begin the dialogue in search of answers to these questions. PMID- 15806507 TI - The ethics of heparin administration to the potential non-heart-beating organ donor. AB - The increasing need and limited sources for organs has stimulated a renewed interest in non-heart-beating organ donation (NHBOD). NHBOD is the donation of organs from cadavers that have been declared dead by cardiopulmonary criteria. Emerging protocols for heparin administration to the potential non-heart-beating organ donor (NHBD) deserve scrutiny. This topic is presented within a limited discussion of organ donation in general. The definition of death has been refined to its present state because of the need for clear parameters in light of a desire to procure organs for transplant. The administration of heparin to the NHBD is intended to prevent the formation of blood clots in the kidneys and liver. Heparin at high doses is clearly not given for the benefit of the patient, and some professionals are concerned that administration may hasten death. Nurses are guided by codes that require the consideration of the ethical principles of autonomy, informed consent, beneficence, and justice while providing compassionate care. An eagerness to procure viable organs for positive transplant outcomes must not be the guiding force in protocols that allow the administration of heparin to the potential NHBD. Heparin administration is supported for these donors within specific parameters. PMID- 15806508 TI - Nursing and genetics: applying the American Nurses Association's Code of Ethics. AB - A professional code of ethics is essential for behaviors to be reinforced in any discipline. The American Nurses Association (ANA) has established a code of ethics for nurses to use as a framework for making ethical decisions with all aspects of health care delivered to the public. With the explosion of genetic discoveries, nurses and clients are facing new ethical dilemmas. It is important for nurses to understand how to use the ANA Code of Ethics when faced with the many complexities of ethical issues involving genetics and health care. This article illustrates how the ANA Code of Ethics can be applied in nursing practice to establish professional behaviors related to ethical issues and genetics. PMID- 15806509 TI - Physical assessment skills: a descriptive study of what is taught and what is practiced. AB - The purpose of this study was to investigate physical assessment skills taught in undergraduate baccalaureate nursing programs and the physical assessment skills used by practicing nurses. Twelve educators teaching physical assessment and 51 practicing nurses were surveyed on 120 physical assessment skills listed in standard nursing textbooks. Median scores were computed on all the skills. The educators taught all of the skills, 92.5% of the skills were taught and practiced in class and 7.5% were taught but not practiced. For the practicing nurses, 37% of the skill were never used, 29% were used on a daily/weekly basis, with 34% used on a monthly/occasional basis. Based on these findings, we recommend that the content of physical assessment courses be reevaluated, with courses focusing on what nurses need to know to practice nursing. PMID- 15806510 TI - Advances in graduate nursing education: beyond the advanced practice nurse. AB - Historically, graduate education in nursing has been primarily concerned with the clinical role. In recent years it has been suggested that graduate education needs to consider alternate programs of study that prepare nurses for clinical leadership that are distinct from management and advanced practice roles. Graduate education is needed that focuses on the skills required to coordinate care and implement outcome-based practice and quality improvement strategies. Two models are currently being proposed that meet these objectives. The first is the population health nurse expert that functions on the macrosystems approach and the second is the clinical nurse leader which is based on a microsystems framework. The two models are compared and a combined model where the clinical nurse leader is based on the population health framework is proposed. PMID- 15806511 TI - Best practices in Web-based courses: generational differences across undergraduate and graduate nursing students. AB - The demand for online courses is greatly increasing across all levels of the curriculum in higher education. With this change in teaching and learning strategies comes the need for quality control to determine best practices in online learning communities. This study examines the differences in student perceptions of the use of technology, educational practices, and outcomes between undergraduate and graduate students enrolled in Web-based courses. The multisite study uses the benchmarking process and the Flashlight Program Evaluating Educational Uses of the Web in Nursing survey instrument to study best practices and examine generational differences between the two groups of students. The outcomes of the study establish benchmarks for quality improvement in online learning. The results support the educational model for online learning and postulates about generational differences for future study. PMID- 15806512 TI - Intestinal and oncocytic variants of pancreatic intraepithelial neoplasia. A morphological and immunohistochemical study. AB - We report 2 previously undescribed morphological variants of pancreatic intraepithelial neoplasia (PanIN). The first variant with an intestinal phenotype was associated with mucinous carcinomas that occurred in the tail of the pancreas of 2 men (60 and 65 years old). The carcinomas lacked the characteristic ovarian like stroma of mucinous cystic neoplasms observed in female patients and did not show a papillary architecture. Whether they represent mucinous cystadenocarcinomas or mucinous carcinomas that arose from the flat variant of intraductal papillary mucinous neoplasms could not be determined with certainty. Microscopically, the intestinal type of PanIN was composed of pseudostratified columnar cells similar to those of colonic adenomas and showing variable degrees of dysplasia. A significant increase in the MIB-1 labeling index correlated with the severity of dysplasia. In contrast to conventional PanIN, the intestinal variant expressed MUC-2 and was MUC-1 negative. The second type of PanIN had an oncocytic phenotype, coexpressed MUC-2 and MUC-1 mucins, and was associated with intraductal oncocytic papillary carcinomas that showed a similar immunohistochemical mucin profile. Both intestinal and oncocytic types of PanIN expressed DPC4 and lacked p53 reactivity. The anatomical separation of the PanINs from the carcinomas and the gradual progression of cytological and architectural abnormalities in both variants of PanIN argue against ductal spread (cancerization of the ducts). The intestinal and oncocytic variants of PanIN broaden the morphological spectrum of this intraductal lesion. Although their significance is unknown, the possibility that these PanIN variants represent cancer precursors should be considered. PMID- 15806513 TI - The value of postmortem examination in cases of metastasis of unknown origin-20 year retrospective data from a tertiary care center. AB - BACKGROUND: Metastasis of unknown origin (MUO) is a diagnostic challenge in clinical practice even with the state of current advanced diagnostic technology. To evaluate the value of autopsy in determining the primary site of MUO, this study reviewed the Hamilton experience-over the last 20 years-with patients autopsied with clinical diagnosis of MUO. METHODS: All autopsy diagnoses from cases performed at the Hamilton Health Sciences Center and St Joseph's Healthcare from 1980 to 2000 were reviewed. Fifty-three cases of MUO were identified (MUO was defined as a patient with pathological and/or radiological diagnosis of a metastatic tumor for which the primary site of malignancy was unknown). The clinical history and gross and microscopic diagnoses for these cases were reviewed. RESULTS: There were 31 men (58.5%) and 22 women (41.5%) in the study. Their mean age was 66 years. Pathological diagnoses at autopsy were adenocarcinoma (n = 37), small cell carcinoma (n = 6), anaplastic carcinoma (n = 3), and undifferentiated carcinoma (n = 3). Primary tumors were identified in 27 patients (51%), most commonly in the lung (n = 8), large bowel (n = 6), and pancreas (n = 4). Histochemical and immunohistochemical stains were helpful in reaching the diagnosis of a primary tumor in 4 of 27 cases. CONCLUSIONS: The following were observed: (1) in this series, autopsy was helpful in establishing the diagnosis of a primary tumor in 51% of the cases, reaffirming the value of postmortem examination in these instances; (2) adenocarcinoma was the most frequent tumor presenting as MUO; (3) the lung and the large bowel were the most frequent sites for primary tumors; and (4) careful gross and histological examinations remain the most important tools in identifying the primary site. PMID- 15806514 TI - The faces of mast cell disease: bone marrow infiltrates in 3 patients with systemic mastocytosis. AB - The clinical spectrum of mast cell disease ranges from relatively innocuous and histologically subtle urticarial skin lesions to an aggressive and fatal leukemic form of mast cell proliferation. Not surprisingly, mast cell infiltrates may show significant microscopic heterogeneity, particularly in the bone marrow, the most common site of involvement in systemic mastocytosis (SM). Herein, 3 cases are presented to illustrate the clinical and morphologic heterogeneity of mast cell disease: the first patient, with long standing urticaria pigmentosa, developed anemia and thrombocytopenia; the second patient presented with a pathologic fracture; and the third patient was suspected to have refractory anemia. Upon bone marrow examination, all 3 patients showed mast cell infiltration with distinct morphologic features and all met the WHO criteria for aggressive systemic mastocytosis. Histochemical methods continue to play a role in the identification of mast cells, with some limitations depending on the degree of differentiation of the mast cells and tissue processing methods. Immunohistochemistry has contributed to the identification of mast cells. Coexpression of CD117 and CD25, as well as expression of the more specific immunohistochemical marker tryptase, is seen in systemic SM. The latter may also be employed as a serum marker in the diagnosis and follow-up of patients with SM. The mast cells, in the majority adults with SM, have somatic point mutations of KIT. PMID- 15806515 TI - Sarcomatoid salivary duct carcinoma. AB - The so-called sarcomatoid salivary duct carcinoma (SSDC) is one of the variants of salivary duct carcinoma (SDC). This neoplasm is characterized by the presence of both a carcinomatous and a sarcomatoid tumor component. The histology and nomenclature of such neoplasms has been a matter of debate for many years. The histologic, immunohistochemical, and electron microscopic findings including those of 4 previously described cases of SSDC are defined and the different attitudes concerning their etiology will be discussed. In addition, the fine needle aspiration biopsy of such a case is presented for the first time. In analogy to typical SDC there seems to be a predilection for elderly men and a location in major salivary glands. The resected SSDC tumors measured between 1.5 and 3.5 cm. Histologically, each case was a composite of SDC and sarcomatoid carcinoma. Immunohistochemical positivity for epithelial membrane antigen (EMA) and cytokeratins (AE1/AE3, CAM 5.2) was shown in the sarcomatoid tumor component. The important cytomorphologic feature of SSDC is the presence of cohesive clusters and flat sheets of cells with a cribriform pattern, in combination with an atypical spindle cell component. The use of the term SSDC seems more appropriate than the term carcinosarcoma , as the immunohistochemical, electron microscopic, and recent molecular findings in this and other biphasic neoplasms imply a monoclonal origin. PMID- 15806516 TI - Optic nerve choristoma. AB - Choristomas are rare lesions that typically involve peripheral nerves and are marked by the presence of histologically normal tissue for an organ or body part other than the site at which it is located. Reports of choristoma involving cranial nerves are extraordinarily rare. We report a case of an optic nerve choristoma arising in a 24-year-old woman. The lesion was marked by a disordered admixture of smooth muscle tissue and optic nerve tissue. The literature on optic nerve choristoma is reviewed. PMID- 15806517 TI - Abdominal lymphangioleiomyomatosis in a man with Klinefelter syndrome: the first reported case. AB - Lymphangioleiomyomatosis (LAM) is an uncommon progressive disease characterized by a hamartomatous tumor-like proliferation of smooth muscle cells that occurs most often in women. This disease commonly involves the lymph nodes, lungs, and mediastinum, and rarely the abdominal sites. We report a case of mesenteric LAM occurring in a 37-year-old man affected by Klinefelter syndrome with a 17-year history of androgen replacement therapy. Histology revealed a hamartomatous proliferation of spindle cells surrounding multiple ectatic lymphatic spaces intermixed with lymphatic follicles. When subjected to immunohistochemical studies, the tumor cells stained positive for muscular markers and negative for estrogen, progesterone, and androgen receptors. The occurrence of LAM in association with Klinefelter syndrome has never been reported in the literature and could represent a further clue in the still unclear pathogenesis of this disease. PMID- 15806518 TI - Uterine epithelioid endometrial stromal sarcoma presenting as a "cervical polyp". AB - Although appearance of conventional uterine endometrial stromal sarcoma is easily recognized on histology, it may uncommonly assume unusual appearances such as uterine tumor resembling ovarian sex-cord tumor, thereby hindering its diagnosis. Recently, its manifestation as an epithelioid neoplasm was described. In this report, we detail yet another instance where this tumor adopted an epithelioid morphology, presenting itself as a polyp extruding from the cervical os in a 41 year-old Chinese woman. Both the polypectomy and subsequent hysterectomy specimens revealed a predominant proliferation of CD10-negative, caldesmon negative, and CD117-positive epithelioid cells set within a stroma containing vascular proliferation resembling endometrial stromal tumor. Areas of typical low grade endometrial stromal sarcoma containing spindle cells that were focally positive for CD10 and negative for CD117 were present in close association with the epithelioid areas. The differential diagnoses and possible implication of CD117 positivity are discussed. PMID- 15806519 TI - Pyogenic granuloma of the sigmoid colon. AB - Pyogenic granuloma (PG) is a polypoid form of lobular capillary hemangioma that usually occurs in the skin and some mucosal surfaces. However, it is extremely rare in the intestinal tract and its preoperative diagnosis is difficult. We report on a case of PG of the sigmoid colon in a 62-year-old woman with a 6-month history of rectal bleeding. A pedunculated polypoid lesion was detected by colonoscopy and a polypectomy was performed. A microscopical study revealed a lobular arrangement proliferation of varying sizes of capillaries within an edematous stroma. A critical review of the English literature yielded only another well-documented case of PG in the large intestine. Diagnostic consideration includes inflammatory polyp and other vascular tumors such as bacillary angiomatosis and the angiomatous variant of Kaposi's sarcoma. Precise recognition of this distinctive vascular neoplasm in the gastrointestinal tract is essential to avoid misdiagnosis and inappropriate treatment. PMID- 15806520 TI - Paraganglioma with ganglioneuromatous component located in the posterior mediastinum. AB - A 4-cm paravertebral mediastinal tumor was resected in a 70-year-old male patient treated for hypertension. The tumor displayed both paraganglioma and ganglioneuroma areas that were in equal proportion and often merged one into the other. Paraganglioma areas contained synaptophysin and chromogranin-positive chief cells and PS100-positive sustentacular cells. Ganglioneuroma areas contained neurofilament-positive mature ganglion cells and PS100-positive Schwann cells. Such pheochromocytoma-ganglioneuroma has not been previously reported in the mediastinum and appears as the adrenal and aorticosympathetic counterpart of gangliocytic paraganglioma described in other anatomic sites. PMID- 15806521 TI - Current status of small peripheral adenocarcinomas of the lung and their importance to pathologists. AB - There has been a large amount of work done recently on small peripheral stage I adenocarcinomas that come to resection. Radiological (including proportion of ground glass opacity) and pathological features of these lesions (predominant bronchioloalveolar component, central scar with or without invasion <0.5 cm) have been shown to be prognostically favorable with cure rate approaching 100% in some series. Most of these studies emanate from Japan. The relevance of these studies to other parts of the world, particularly North America, is discussed in light of the fact that some recent chemotherapeutic studies with gefitinib have shown increased response in individuals of Asian origin, suggesting that some genetic differences may be significant. The relevance of these findings to pathologists and the pathological study of small peripheral adenocarcinomas from elsewhere in the world are discussed. PMID- 15806523 TI - Cultural competence: a call to action. PMID- 15806525 TI - Perianesthesia nursing at an ophthalmic hospital in the Middle East. AB - Health care delivery differs from the United States among different cultures and countries. This descriptive article describes health care delivery at two ophthalmic hospitals in a Middle Eastern country. This description may not be representative of health care delivery throughout that country but is limited to the hospitals described. The purpose of this article is two-fold. First, it allows the readers to compare and contrast health care delivery to their own setting and practice. Second, it allows the reader to appreciate the tremendous work being done in a difficult environment. PMID- 15806526 TI - Strategies to reduce delays in admission into a postanesthesia care unit from operating rooms. AB - The authors performed a systematic review of strategies to reduce delays in admission into PACUs from ORs. The purpose of this article was to evaluate for managers how to choose interventions based on effectiveness and practicality. The authors discuss optimization methods that can be used to sequence consecutive cases in the same OR, by the same surgeon, on the same day, based on the objective of reducing delays in PACU admission due to the unavailability of unfilled PACU beds. Although effective, such methods can be impractical because of large organizational change required and limited equipment or personnel availability. When all physical beds are not full, PACU nurse staffing can be adjusted. Statistical methods can be used to ensure that nursing schedules closely match the times that minimize delays in PACU admission. These methods are effective and practical. Explicit criteria can be applied to assist in deciding when to assign other qualified nurses to the PACU, when to ask PACU nurses to work late, and/or when to make a decision on the day before surgery to add more PACU nurses for the next day (if available). The latter would be based on statistical forecasts of the timing of patients' admissions into the PACU. Whether or not all physical beds are full, the risk of delays in PACU admission is relatively insensitive to economically feasible reductions in PACU length of stay. Such interventions should be considered only if statistical analysis, performed by using computer simulation, has established that reducing PACU length of stay will reduce delays in admission at a manager's facility. PMID- 15806527 TI - Perianesthesia nursing considerations for Ganz periacetabular osteotomy patients. AB - Various approaches have been proposed, developed, and implemented for patients suffering from chronic and sometimes incapacitating hip pain. This article explores the history of surgical procedures for hip pain related to developmental dysplasia, with a focus on the Ganz periacetabular osteotomy. Finally, some future approaches that may offer ways to improve the results of periacetabular osteotomies are explored. PMID- 15806528 TI - Whole lung lavage in the treatment of pulmonary alveolar proteinosis. AB - Although a rare condition, pulmonary alveolar proteinosis (PAP) can be a very devastating diagnosis with life-altering consequences. This case study follows the path of a young woman who is currently undergoing whole lung lavage as treatment for pulmonary alveolar proteinosis. The entire concept of flooding a lung with large quantities of saline as a treatment for lung disease is contrary to normal respiratory care. Caring for the patient with PAP provides many challenges for the perianesthesia nurse. Management of the postanesthesia airway, oxygen administration and maintenance of oxygen saturation, and pain relief skills are all of high importance to the patient with PAP. These skills plus the emotional support provided by the experienced perianesthesia nurse can ensure a safe recovery from this unusual procedure. PMID- 15806529 TI - Anne Allen: the blue streak. PMID- 15806530 TI - Prevention of anesthesia awareness. PMID- 15806531 TI - Development of a preoperative admission group. PMID- 15806532 TI - Inflammation II: sepsis. PMID- 15806533 TI - The use of opioids in the postoperative setting: focus on morphine, hydromorphone, and fentanyl. PMID- 15806534 TI - SurgiChip--new technology for prevention of wrong site, wrong procedure, wrong person surgery. PMID- 15806537 TI - Transition: a metamorphosis of nursing? PMID- 15806538 TI - Active sites on oxide surfaces: ZnO-catalyzed synthesis of methanol from CO and H2. PMID- 15806539 TI - Chemotherapy and attentional dysfunction in survivors of childhood acute lymphoblastic leukemia: effect of treatment intensity. AB - BACKGROUND: Central nervous system (CNS) directed chemotherapy is replacing prophylactic cranial irradiation in treatment protocols for childhood acute lymphoblastic leukemia (ALL), mainly to reduce long-term neuropsychological sequelae. We evaluated the effects of chemotherapy on attentional function in survivors of ALL and examined whether possible deficits are related to treatment intensity. METHODS: In a multi-center study, we compared attentional function in 36 children at least 1 year after finishing treatment with chemotherapy only for ALL, with a cancer control group consisting of 39 Wilms tumor patients and with 110 healthy children. We differentiated between standard- and intensified ALL treatment. The role of previously reported risk factors for neuropsychological deficits was also assessed. RESULTS: After chemotherapy, attentional deficits were detected in patients with ALL, but not in Wilms tumor patients. Children treated according to standard ALL protocols performed worse than healthy controls on only 1 of 10 outcome measures (P = 0.004), while those who had received intensified treatment performed worse on four outcome measures (0.0001 < P < 0.004). Higher treatment intensity, young age at diagnosis, and female gender were associated with worse performance. CONCLUSIONS: CNS-directed chemotherapy, even in the absence of cranial irradiation, is associated with attentional dysfunction in survivors of childhood ALL, particularly in case of intensified treatment protocols. These sequelae stress the importance of reducing doses of neurotoxic chemotherapy as much as possible in the design of future treatment protocols for ALL. PMID- 15806540 TI - Polymorphism of angiotensin converting enzyme is associated with severe circulatory compromise in febrile neutropenic children with cancer. AB - Angiotensin converting enzyme (ACE) gene insertion(I)/deletion(D) polymorphism influences the outcome of a number of cardiovascular diseases. ACE I/D polymorphism was investigated by PCR in 207 pediatric cancer patients and 144 controls. ACE I/D distribution of patients and controls was similar. The frequency of the D allele and the prevalence of the deletion (DD) genotype were significantly (P < 0.05) higher among patients with severe circulatory compromise requiring treatment in the intensive care unit (ICU) than among the other patients and controls. Patients with the DD and ID genotypes spent significantly (P < 0.05) longer time in the ICU than patients with the II genotype. PMID- 15806541 TI - Symptoms of post-traumatic stress in children with cancer and their parents: effects of informant and time from diagnosis. AB - BACKGROUND: Post-traumatic stress disorder (PTSD) has been put forth as a model for understanding the adjustment of children with cancer and their parents, but findings in the literature regarding the prevalence of post-traumatic stress symptoms (PTSS) have been mixed. This study examined PTSS levels in both patients and parents as a function of time elapsed from diagnosis, and by use of parent versus child reports for assessing patient PTSS. PROCEDURE: Four groups of patients (total N = 162) differing in time elapsed since diagnosis were assessed with standardized measures of PTSS. Patients reported on their own symptoms, while parents reported on their symptoms as well as symptoms in their child. RESULTS: Significant differences between groups were observed on all patient PTSS measures. Recently diagnosed patients obtained higher PTSS scores than survivors more than 5 years from diagnosis. Likewise, parents of recently diagnosed patients self-reported significantly higher levels of PTSS than parents of long term survivors. Parent-child correlations were high and no differences were observed between parent and patient report of patient PTSS. Comparison with historical control data suggests that levels of PTSS in patients are very low. CONCLUSION: Concordance between parent and child reports of child PTSS suggests that data may be obtained with reasonable confidence from either if only one informant is available. Higher levels of PTSS in patients who are recently diagnosed (and their parents) in comparison to long-term survivors, suggest that the symptoms reported reflect primarily a concurrent response to ongoing acute stressors, rather than a post-traumatic re-experiencing of past traumas. This, in combination with the low levels of patient PTSS raise questions regarding the utility of PTSD as a model for understanding patient and parent adjustment to childhood cancer. PMID- 15806542 TI - Intramedullary spinal cord astrocytomas in children. PMID- 15806543 TI - Allogeneic graft-versus-hepatoblastoma effect. AB - Although the survival rate for pediatric patients with hepatoblastoma has improved, prognosis is still poor when the disease is unresectable and refractory to chemotherapy. Therefore, novel approaches are warranted. Herein, we describe a patient with recurrent metastatic hepatoblastoma who received a non-myeloablative hematopoietic stem cell transplantation from an HLA-matched unrelated donor. After withdrawal of immunosuppressant and establishment of full donor T-cell engraftment, the tumor regressed and serum alpha-fetoprotein level decreased in concurrence with the onset of graft-versus-host disease (GVHD). Her disease recurred when GVHD resolved. This patient's clinical course provides evidence for the probable existence of allogeneic graft-versus-hepatoblastoma effect. PMID- 15806544 TI - Randomized comparison of antibiotics with and without granulocyte colony stimulating factor in children with chemotherapy-induced febrile neutropenia: a report from the Children's Oncology Group. AB - PURPOSE: To determine if granulocyte colony-stimulating factor (G-CSF) with empirical antibiotics accelerates febrile neutropenia resolution compared with antibiotics without it. PATIENTS AND METHODS: Eligible children were treated without prophylactic G-CSF and presented with fever (temperature >38.3 degrees C) and neutropenia afterward. Patients with acute myelogenous leukemia and myelodysplastic syndrome were excluded. Assignments were randomized between G-CSF (5 microg/kg/day) or none beginning within 24 hr of antibiotics. Subcutaneous administration was recommended, but intravenous G-CSF was allowed. Patients remained on study until absolute neutrophil count (ANC) >500/microl and > or =48 hr without fever. RESULTS: One of 67 patients enrolled was ineligible, 59 had acute lymphoblastic leukemia (ALL). Thirty-four were assigned to antibiotics, 32 to G-CSF plus antibiotics. Adding G-CSF significantly reduced neutropenia and febrile neutropenia recovery times. Median days to febrile neutropenia resolution was nine earlier with G-CSF (4 vs. 13 days) (P < 0.0001). However, there was no difference in the resolution of fever between arms. Hospitalization median was shorter by 1 day with G-CSF (4 vs. 5 days) (P = 0.04). There was no difference in the duration of IV and oral antibiotic treatment, addition of antifungal therapy, and shock incidence. A trend for decreased incidence of late fever with G-CSF was noted (6.3 vs. 23.5%) (P = 0.08). CONCLUSIONS: Adding G-CSF to empiric antibiotic coverage accelerates chemotherapy-induced febrile neutropenia resolution by 9 days in pediatric patients, mainly with ALL, which results in a small but significant difference in the median length of hospitalization. PMID- 15806545 TI - AMOR: a proposed cooperative effort to improve outcomes of childhood cancer in Central America. AB - The dramatic reduction of pediatric cancer mortality rates has been one of the greatest accomplishments of contemporary medicine. About 80% of children with cancer are now expected to be cured by current therapies. However, most of the world's children have no access to cancer treatment. The translation of effective pediatric cancer therapies to impoverished regions of the world presents an enormous challenge to the health care profession. Over the past 20 years, efforts have been under way to extend adequate cancer treatment to an increasing number of children in developing countries. These initiatives, collectively designated "twinning programs," consist essentially of a partnership between a pediatric cancer unit in a developing country and a group of health care providers in the developed world. Here we review the twinning programs that have been implemented in Central America, discuss their impact on the development of local resources and the outcome of childhood cancer, and propose a collaborative research initiative aimed at improving the international dissemination of progress in pediatric hematology-oncology. PMID- 15806546 TI - Outcome of CNS and pulmonary enteroviral infections after hematopoietic cell transplantation. PMID- 15806547 TI - Glucose metabolism at high density growth of E. coli B and E. coli K: differences in metabolic pathways are responsible for efficient glucose utilization in E. coli B as determined by microarrays and Northern blot analyses. AB - In a series of previous reports it was established by implementing metabolic flux, NMR/MS, and Northern blot analysis that the glyoxylate shunt, the TCA cycle, and acetate uptake by acetyl-CoA synthetase are more active in Escherichia coli BL21 than in Escherichia coli JM109. These differences were accepted as the reason for the differences in the glucose metabolism and acetate excretion of these two strains. Examination of the bacterial metabolism by microarrays and time course Northern blot showed that in addition to the glyoxylate shunt, the TCA cycle and the acetate uptake, other metabolic pathways are active differently in the two strains. These are gluconeogenesis, sfcA shunt, ppc shunt, glycogen biosynthesis, and fatty acid degradation. It was found that in E. coli JM109, acetate is produced by pyruvate oxidase (poxB) using pyruvate as a substrate rather than by phosphotransacetylase-acetate kinase (Pta-AckA) system which uses acetyl-CoA. The inactivation of the gluconeogenesis enzyme phosphoenolpyruvate synthetase (ppsA), the activation of the anaplerotic sfcA shunt, and low and stable pyruvate dehydrogenase (aceE, aceF) cause pyruvate accumulation which is converted to acetate by pyruvate oxidase B. The behavior of the ppsA, acs, and aceBAK in JM109 was dependent on the glucose supply strategy. When the glucose concentration was high, no transcription of these genes was observed and acetate concentration increased, but at low glucose concentrations these genes were expressed and the acetate concentration decreased. It is possible that there is a major regulatory molecule that controls not only ppsA and aceBAK but also acs. The gluconeogenesis pathway (fbp, pckA, and ppsA) which leads to glycogen accumulation is constitutively active in E. coli BL21 regardless of glucose feeding strategy. PMID- 15806548 TI - Chain-growth polycondensation for well-defined condensation polymers and polymer architecture. AB - The historical development of our research on polycondensation that proceeds in a chain-growth polymerization manner ("chain-growth polycondensation") for well defined condensation polymers is described. We first studied polycondensation in which change of the substituent effect induced by bond formation drove the reactivity of the polymer end group higher than that of the monomer. In this approach, well-defined aromatic polyamides, polyesters, polyethers, and poly(ether sulfone)s were obtained. The second approach was the study of the phase-transfer polymerization of a solid monomer dispersed in an organic solvent. In this type of polymerization, the solid monomer was physically unable to react with another monomer and was carried with the phase transfer catalyst into the solution phase where it reacted with an initiator and the polymer end group in the solvent in a chain polymerization manner. We also found catalyst-transfer polycondensation as a third approach to chain-growth polycondensation. In the Ni catalyzed polycondensation of 2-bromo-5-chloromagnesiothiophenes, the Ni catalyst transferred to the polymer end group, and a coupling reaction occurred there to yield a well-defined polythiophene. This chain-growth polycondensation was applied to the synthesis of condensation polymer architectures such as block copolymers, star polymers, graft copolymers, and so on. PMID- 15806549 TI - Medium optimization for hen egg white lysozyme production by recombinant Aspergillus niger using statistical methods. AB - Statistics-based experimental design was used to investigate the effect of medium components (starch, peptone, ammonium sulfate, yeast extract, and CaCl2.2H2O) on hen's egg white lysozyme production by Aspergillus niger HEWL WT-13-16. A 2(5-1) fractional factorial design augmented with center points revealed that peptone, starch, and ammonium sulfate were the most significant factors, whereas the other factors were not important within the levels tested. The method of steepest ascent was used to approach the proximity of optimum. This task was followed by a central composite design to develop a response surface for medium optimization. The optimum medium composition for lysozyme production was found to be: starch 34 g L-1, peptone 34 g L-1, ammonium sulfate 11.9 g L-1, yeast extract 0.5 g L-1, and CaCl2.2H2O 0.5 g L-1. This medium was projected to produce, theoretically, 212 mg L-1 lysozyme. Using this medium, an experimental maximum lysozyme concentration of 209+/-18 mg L-1 verified the applied methodology. PMID- 15806550 TI - Comparing the function of the corticospinal system in different species: organizational differences for motor specialization? AB - An appreciation of the comparative functions of the corticospinal tract is of direct relevance to the understanding of how results from animal models can advance knowledge of the human motor system and its disorders. Two critical functions of the corticospinal tract are discussed: first, the role of descending projections to the dorsal horn in the control of sensory afferent input, and second, the capacity of direct cortico-motoneuronal projections to support voluntary execution of skilled hand and finger movements. We stress that there are some important differences in corticospinal projections from different cortical regions within a particular species and that these projections support different functions. Therefore, any differences in the organization of corticospinal projections across species may well reflect differences in their functional roles. Such differences most likely reflect features of the sensorimotor behavior that are characteristic of that species. Insights into corticospinal function in different animal models are of direct relevance to understanding the human motor system, providing they are interpreted in relation to the functions they underpin in a given model. Studies in non-human primates will continue to be needed for understanding special features of the human motor system, including feed-forward control of skilled hand movements. These movements are often particularly vulnerable to neurological disease, including stroke, cerebral palsy, movement disorders, spinal injury, and motor neuron disease. PMID- 15806551 TI - Functional sparing of intrafusal muscle fibers in muscular dystrophies. AB - In a previous study, we showed that patients with muscular dystrophies (MDs) perceive passive movements, experience sensations of illusory movement induced by muscle tendon vibration, and have proprioceptive-regulated sways in response to vibratory stimulation applied to the neck and ankle muscle tendons. These findings argue for preserved proprioceptive functions of muscle spindles. However, it is unclear whether the function of intrafusal muscle fibers is spared, i.e., whether they retain their ability to contract when submitted to a fusimotor drive. To answer this question, we analyzed the effects of reinforcement maneuvers (mental computation and the Jendrassik maneuver) that are known to increase muscle spindle sensitivity via fusimotor drive in healthy subjects. Nine patients with different MDs participated in the study. Reinforcement maneuvers increased both the mean amplitude of the Achilles tendon reflex (187 +/- 52.9% of the mean control amplitude) and the sensitivity of muscle spindle afferents to imposed movements of the ankle. The same reinforcement maneuvers failed to alter the amplitude of the Hoffmann reflex in the triceps surae muscle. These results suggest that the intrafusal muscle fibers preserve their contractile abilities in slowly progressive MDs. The reasons for a differential impairment of intrafusal and extrafusal muscle fibers and the clinical implications of the present results are discussed. PMID- 15806552 TI - Involvement of superficial peroneal sensory nerve in common peroneal neuropathy. AB - Motor involvement in common peroneal neuropathy (CPN) frequently shows a selective pattern with regard to deep and superficial divisions of the peroneal nerve, by clinical examination and needle electromyography. The involvement of the sensory branch of the superficial peroneal nerve (SPN) has not been well established using nerve conduction studies. Among 42 cases of electrophysiologically defined CPN in 35 patients, 37 (88%) had clinical or electrophysiologic evidence for SPN involvement, but only 20 had diminished superficial peroneal sensory response (SPSR) amplitudes. Many of the cases involving normal SPSRs had significant axonal loss involving deep peroneal motor fibers. Sparing of the superficial peroneal sensory fibers provides further evidence for the selective vulnerability of different nerve fascicles to injury. This is an important pattern to recognize; from a practical standpoint, focal segmental conduction abnormalities in the motor nerve and EMG findings can help to differentiate these lesions from L-5 radiculopathy. PMID- 15806553 TI - Interaction of the conserved meiotic regulators, BOULE (BOL) and PUMILIO-2 (PUM2). AB - Germ cell development is complex; it encompasses specification of germ cell fate, mitotic replication of early germ cell populations, and meiotic and postmeiotic development. Meiosis alone may require several hundred genes, including homologs of the BOULE (BOL) and PUMILIO (PUM) gene families. Both BOL and PUM homologs encode germ cell specific RNA binding proteins in diverse organisms where they are required for germ cell development. Here, we demonstrate that human BOL forms homodimers and is able to interact with a PUMILIO homolog, PUM2. We mapped the domain of BOL that is required for dimerization and for interaction with PUM2. We also show that BOL and PUM2 can form a complex on a subset of PUM2 RNA targets that is distinct from targets bound by PUM2 and another deleted in azoospermia (DAZ) family member, DAZ-like (DAZL). This suggests that RNA sequences bound by PUM2 may be determined by protein interactions. This data also suggests that although the BOL, DAZ, and DAZL proteins are all members of the same gene family, they may function in distinct molecular complexes during human germ cell development. PMID- 15806554 TI - Synthetic examination of incorrectly proposed structures of biomolecules. AB - Many incorrect structures of biomolecules have been proposed for natural products. Synthesis of compounds having the proposed structures often enabled us to judge the correctness of the proposals. In some cases, we were able to revise the structures by synthesizing the biomolecules themselves. In other cases, we were able to definitely disprove the proposed structures. Some examples treated in this review include: auxin-a and b; the sex pheromone of Chlamydomonas; sex pheromones of the gypsy moth, the American cockroach, and the pink bollworm moth; Persoons' periplanone-A; orobanchol; naurol A; bifurcarenone; koninginin A; alpha acoradiene; himachalene-type pheromones of the flea beetle; differolide; blattellastanoside A and B; etc. PMID- 15806555 TI - Vine-twining polymerization: a new preparation method for well-defined supramolecules composed of amylose and synthetic polymers. AB - In this article we describe a new method of polymerization called "vine-twining polymerization" for preparation of well-defined supramolecules, which are amylose polymer inclusion complexes. The method was achieved by enzymatic polymerization of alpha-D-glucose-1-phosphate catalyzed by phosphorylase in the presence of various synthetic polymers such as polyethers, polyesters, poly(ester-ether), and amphiphilic block copolymer. Powder X-ray diffraction (XRD) and 1H-NMR measurements determined the structures of the products to be inclusion complexes. The XRD patterns were completely different from those of amylose and guest polymers. The 1H-NMR spectra of the products indicated that the structures were composed of amylose and guest polymers. The formation process of the inclusion complexes during the enzymatic polymerization was also evaluated. In addition, we revealed that the bulkiness of the end groups and the hydrophobicity of the guest polymers strongly affected the formation of the inclusion complexes. By means of this method of polymerization, a graft polymer having inclusion complexes as side chains was prepared. Furthermore, as an evolution of the "vine-twining polymerization," we attempted a system of parallel polymerization to form an inclusion complex of amylose with a strongly hydrophobic guest polymer. PMID- 15806556 TI - The analysis of telomere length and telomerase activity in cloned pigs and cows. AB - Inefficiency in the production of cloned animals is most likely due to epigenetic reprogramming errors after somatic cell nuclear transfer (SCNT). In order to investigate whether nuclear reprogramming restores cellular age of donor cells after SCNT, we measured telomere length and telomerase activity in cloned pigs and cattle. In normal pigs and cattle, the mean telomere length was decreased with biological aging. In cloned or transgenic cloned piglets, the mean telomere length was elongated compared to nuclear donor fetal fibroblasts and age-matched normal piglets. In cloned cattle, no increases in mean telomere length were observed compared to nuclear donor adult fibroblasts. In terms of telomerase activity, significant activity was observed in nuclear donor cells and normal tissues from adult or new-born pigs and cattle, with relatively higher activity in the porcine tissues compared to the bovine tissues. Cloned calves and piglets showed the same level of telomerase activity as their respective donor cells. In addition, no difference in telomerase activity was observed between normal and transgenic cloned piglets. However, increased telomerase activity was observed in porcine SCNT blastocysts compared to nuclear donor cells and in vitro fertilization (IVF)-derived blastocysts, suggesting that the elongation of telomere lengths observed in cloned piglets could be due to the presence of higher telomerase activity in SCNT blastocysts. In conclusion, gathering from the comparative studies with cattle, we were able to demonstrate that telomere length in cloned piglets was rebuilt or elongated with the use of cultured donor fetal fibroblasts. PMID- 15806557 TI - Production of alpha 1,3-galactosyltransferase gene knockout pigs expressing both human decay-accelerating factor and N-acetylglucosaminyltransferase III. AB - Heterozygous alpha 1,3-galactosyltransferase (GT) gene knockout pigs were produced with transgenic pig fetal cells expressing both human decay-accelerating factor (hDAF) and N-acetylglucosaminyltransferase III (GnT-III). In this study, we assessed the gene targeting efficiency in the transgenic pig fetal cells derived from different fetal tissues such as brain, skin, heart, and liver, or fetal carcass. Targeted cell colonies were selected by hygromycin B. The GT knockout colonies (KO colonies) were obtained equally from the cells derived from all tissues except liver. Staining with five antibodies against intermediate filaments, all examined KO cell lines stained positive for vimentin with the exception of a colony that stained positive for both vimentin and glial fibrillary acidic protein simultaneously. This is the first study to produce KO cells from the astrocytes. Some of these KO cell lines were used for nuclear transfer (NT) to obtain KO pig fetuses. Fourteen fetuses were obtained from two recipients of the embryo transfer and eight of them had normal ploidy. The cells from the KO pig fetuses were also used for NT to produce cloned KO pigs. Two healthy clone pigs were born. These pigs were determined to have a heterozygous knockout GT gene and the two transgenes. The cells collected from the KO pigs were shown to have similar expression levels of hDAF and GnT-III compared to their original transgenic pigs and less than a half levels of the alphaGal epitopes existed in wild-type pig cells. PMID- 15806559 TI - Unequal segregation of parental chromosomes in embryonic stem cell hybrids. AB - Chromosome segregation was studied in 14 intra- and 20 inter-specific hybrid clones generated by fusion of Mus musculus embryonic stem (ES) cells with fibroblasts or splenocytes of DD/c mice or Mus caroli. As a control for in vitro evolution of tetraploid karyotype we used a set of hybrid clones obtained by fusion of ES cells (D3) with ES cells (TgTP6.3). Identification of the parental chromosomes in the clones was performed by microsatellite analysis and in situ hybridization with labeled species-specific probes. Both analyses have revealed three types of clones: (i) stable tetraploid, observed only for ES x ES cell hybrids; (ii) bilateral loss of chromosomes of both ES and somatic partners; (iii) unilateral segregation of chromosomes of the somatic partner. Observed unilateral segregation was extensive in ES-splenocyte cell hybrids, but lower in ES-fibroblast hybrid clones. Developmental state of the somatic partner is presumably responsible for directional chromosome loss. Nonrandom segregation implies that initial differences in the parental homologous chromosomes were not immediately equalized implying at least transient persistence of the differentiated epigenotype. PMID- 15806558 TI - Optimization of real time RT-PCR methods for the analysis of gene expression in mouse eggs and preimplantation embryos. AB - This study was carried out to optimize conditions for using real time RT-PCR as an efficient and precise quantitative method for estimating the transcript levels of genes expressed in samples containing miniscule amounts of RNA, such as single mammalian oocytes and embryos. First, using mouse eggs and blastocysts, we tested three kinds of RNA isolation or collection methods: TRIZOL reagent, oligo-dT conjugated beads, or three freeze/thaw cycles with the reverse transcription buffer. There were no significant differences among three groups in mRNA quantity as assayed by real time RT-PCR analysis. Second, we compared the efficacy of real time analysis between TaqMan fluorescent probes and the SYBR-green dye system. The two systems presented similar real time RT-PCR profiles for the 16s ribosomal protein gene from oocytes to blastocysts. Third, RNA from mouse embryos at defined stages of preimplantation development were isolated and the levels of transcripts encoded by several housekeeping genes (GAPDH, beta-actin, ribosomal protein L7, 16s ribosomal protein, histone H2A.Z) were quantitatively analyzed by real time RT-PCR. The histone H2A.Z and 16s ribosomal protein slightly increased from the egg to blastocyst stages by approximately 10- and 30-fold, respectively. However, other transcripts increased more than 300-fold as a function of developmental stage from eggs to blastocysts. Our results suggest that the simple freezing/thawing method for RNA collection, the economic SYBR-green dye system, and histone H2A.Z gene as an internal control should be useful for the real time RT-PCR analysis of single mouse eggs and preimplantation embryos. PMID- 15806560 TI - Impact of pro-oxidant agents on the morula-blastocyst transition in bovine embryos. AB - Exposing day 5 bovine morulae to reactive oxygen species induces a delayed degeneration of some blastocysts on day 8 post-insemination (pi) but without affecting the blastocyst rates. The aim of this study was to characterize the resisting and the degenerating population of blastocysts. The kinetics of degeneration of the embryos exposed to the two pro-oxidant agents: 2,2'-azobis (2 amidinopropane) dihydrochloride (AAPH) and buthionine sulfoximine (BSO) was evaluated using time-lapse cinematography. With both agents the first signs of degeneration appeared at day 7.5 pi but the duration of the degeneration process was shorter in presence of AAPH than BSO (4.2 vs. 12.5 hr, ANOVA, P < 0.05). The resisting blastocysts derived from morulae with a larger diameter (mean diameter: 161 vs. 154 microm, ANOVA, P < 0.05) and showed an earlier cavitation (135 vs. 142 hpi, P < 0.05) than the degenerating ones. The profile of protein neosynthesis at day 7 was not affected by the treatment. The proportion of male embryos was more important in the resisting than in the degenerating population (70 vs. 55%, chi2, P < 0.05) especially when the stress was induced by AAPH. The quality of the resisting embryos, measured by the total cell number and the rate of apoptosis, did not seem to be affected when compared to control embryos. In conclusion, resistance to oxidative stress seems related to the kinetics of development and/or the sex of the embryos. Resisting embryos apparently display a quality similar to untreated embryos. PMID- 15806561 TI - Effects of duration, concentration, and timing of ionomycin and 6 dimethylaminopurine (6-DMAP) treatment on activation of goat oocytes. AB - The protocol of ionomycin followed by 6-dimethylaminopurine (6-DMAP) is commonly used for activation of oocytes and reconstituted embryos. Since numerous abnormalities and impaired development were observed when oocytes were activated with 6-DMAP, this protocol needs optimization. Effects of concentration and treatment duration of both drugs on activation and development of goat oocytes were examined in this study. The best oocyte activation (87-95%), assessed by pronuclear formation, was obtained when oocytes matured in vitro for 27 hr were treated with 0.625-20 microM ionomycin for 1 min before 6-hr incubation in 2 mM 6 DMAP. Progressional reduction of time for 6-DMAP-exposure showed that the duration of 6-DMAP treatment can be reduced to 1 hr from the second up to the fourth hour after ionomycin, to produce activation rates greater than 85%. Activation rates of oocytes in vitro matured for 27, 30, and 33 hr were higher (P < 0.05) than that of oocytes matured for 24 hr when treated with ionomycin plus 1 hr (the third hour) 6-DMAP, but a 4-hr incubation in 6-DMAP enhanced activation of the 24-hr oocytes. Goat activated oocytes began pronuclear formation at 3 hr and completed it by 5-hr post ionomycin. An extended incubation in 6-DMAP (a) impaired the development of goat parthenotes, (b) quickened both the release from metaphase arrest and the pronuclear formation, and (c) inhibited the chromosome movement at anaphase II (A-II) and telophase II (T-II), leading to the formation of one pronucleus without extrusion of PB2. In conclusion, duration, concentration, and timing of ionomycin and 6-DMAP treatment had marked effects on goat oocyte activation, and to obtain better activation and development, goat oocytes matured in vitro for 27 hr should be activated by 1 min exposure to 2.5 microM ionomycin followed by 2 mM 6-DMAP treatment for the third hour. PMID- 15806562 TI - Topological polymer chemistry by dynamic selection from electrostatic polymer self-assembly. AB - A collection of recent developments in topological polymer chemistry is presented. First, topological isomerism occurring on randomly coiled, flexible polymer molecules having cyclic and linear structures is discussed. Second, an electrostatic self-assembly and covalent fixation strategy has been developed for the synthesis of polymeric topological isomers. These isomers have double cyclic, manacle-, and theta-shaped constructions, and are prepared by using either linear or star telechelic polymer precursors having moderately strained cyclic ammonium salt groups, which carry multifunctional carboxylate counteranions. A technique of reversed-phase chromatography (RPC) is demonstrated as an effective means to separate polymers with different topologies, especially polymeric topological isomers. A further extension of topological polymer chemistry has been observed by dynamic selection from electrostatic polymer self-assembly to enable the effective formation of tadpole-shaped, cyclic-linear hybrid topologies. PMID- 15806563 TI - Nitroxide-mediated radical processes. AB - In the present short review article recent achievements in nitroxide-mediated radical polymerizations are presented. The basic concept behind these reactions, which is the Persistent Radical Effect (PRE), will be briefly explained. The effect of the nitroxide structure on the polymerization process will be discussed. Moreover, results of nitroxide-mediated radical polymerizations in aqueous dispersions will be summarized. Finally, applications of the PRE to environmentally benign radical chemistry such as nitroxide-mediated alkoxyamine isomerization and carboaminoxylation reactions are presented. Moreover, the potential use of microwave-induced heating to conduct these thermal radical reactions will be discussed. PMID- 15806564 TI - Thermomechanics of the shape memory effect in polymers for biomedical applications. AB - We examine the shape memory effect in polymer networks intended for biomedical, and specifically cardiovascular, applications. The polymers were synthesized by photopolymerization from a tert-butyl acrylate monomer with a diethyleneglycol diacrylate crosslinker. Three-point flexural tests were used to systematically investigate the thermomechanics of shape storage (predeformation) and shape recovery. The glass transition temperature, T(g), of the polymers was determined to be approximately 65 degrees C. The polymers show 100% strain recovery, at low and high predeformation temperatures, up to maximum strains of approximately 80%. The polymers show a sigmoidal free strain recovery response as a function of increasing temperature at a constant heating rate. Free strain recovery was determined to depend on the temperature during predeformation; lower predeformation temperatures (T < T(g)) decreased the temperature required for free strain recovery. Constrained stress recovery shows a complex evolution as a function of temperature and also depends on the temperature during predeformation. Stress recovery after low-temperature predeformation (T < T(g)) shows a peak in the generated recovery stress, whereas stress recovery after high temperature predeformation (T > T(g)) is sigmoidal. The isothermal free strain recovery rate was found to increase with increasing temperature or decreasing predeformation temperature. The thermomechanical results are discussed in light of potential biomedical applications, and a prototype device is presented. PMID- 15806566 TI - Decomposition and long-lasting downregulation of extracellular matrix in perineuronal nets induced by focal cerebral ischemia in rats. AB - The upregulation of extracellular matrix components, especially chondroitin sulfate proteoglycans, after brain injury and stroke is known to accompany the glial reaction, forming repellent scars that hinder axonal growth and the reorganization of the injured neuronal networks. The extracellular matrix associated with perineuronal nets (PNs) in the primarily injured and remote regions has not yet been systematically analyzed. We use the model of permanent middle cerebral artery occlusion (MCAO) to investigate the acute and long-lasting consequences of ischemia for PNs, related to the damage of neurons and reactions of glial cells, in spontaneously hypertensive rats. Extracellular matrix components associated with PNs around cortical interneurons and neurons in thalamic nuclei were characterized 1, 7, 14, and 35 days after MCAO, using Wisteria floribunda agglutinin (WFA) staining and immunocytochemistry. The degradation of PNs in the infarct core was initiated by loss of WFA-binding matrix components, indicating the cleavage of glycosaminoglycan chains of chondroitin sulfate proteoglycans. Immunostaining showed the subsequent removal of proteoglycan core proteins within the extending microglia/macrophage invasion zone lasting for 2 weeks after MCAO. In the cortical periinfarct region, delineated by an astrocytic scar against the infarct core, the number of WFA stained and proteoglycan core protein-immunoreactive PNs was permanently reduced. In the homolateral ventroposterior thalamus, the delayed decrease in perineuronal matrix was related to the distribution pattern of activated microglia and massive neuronal degeneration. It can be concluded from these results that complementary to the known upregulation of matrix components in the glial scar, deficits in the expression of the neuron-associated extracellular matrix develop in the periinfarct and remote regions. These deficits may contribute to the long-lasting functional impairments after stroke. PMID- 15806565 TI - Interactions between self-assembled polyelectrolyte shells and tumor cells. AB - Layer-by-layer self-assembled polyelectrolyte shells are a new class of micro/nanocapsules with unique physicochemical properties for potential applications in drug/gene delivery. The objective of this study was to investigate the interactions of polyelectrolyte shells ( approximately 1 mum in diameter) with MCF-7 breast cancer cells and identify key parameters that affect such interactions. Tailoring of surface properties of polyelectrolyte shells was achieved by choosing different outermost layer materials, including cationic polymers, anionic polymers, and lipid bilayers. Different surface compositions led to a wide range of electrostatic potentials from -46 to +47 mV in phophate buffered saline buffer. Confocal microscopy studies showed that the polyelectrolyte shells were internalized into the cell cytoplasm, but not into the nuclei. Correlation of cell uptake with shell surface compositions was complicated by the adsorption of serum proteins on the surface of polyelectrolyte shells, particularly polycation-coated shells. To prevent protein adsorption, poly(ethylene glycol) (PEG) grafted poly(ethyleneimine) (PEI) copolymers (1:1, 1:5, 1:10 graft ratios) were synthesized and introduced on the shell surface. Shells coated with PEI-PEG copolymers effectively reduced protein adsorption whereas PEI-PEG copolymers with lower graft ratios achieved higher cell uptake efficiency after 24 h of incubation with MCF-7 cells. PMID- 15806567 TI - Three-dimensional stereotactic surface projection SPECT analysis in Parkinson's disease with and without dementia. AB - We investigated regional cerebral blood flow (rCBF) using three-dimensional stereotactic surface projection (3D-SSP) analysis in 30 patients initially diagnosed as Parkinson's disease (PD), and compared differences in rCBF between patients with and without PD-related manifestations. 3D-SSP analysis of cerebral perfusion was performed by use of a control database. Compared to age-matched controls, there were multiple hypoperfusion areas in cases where the original diagnosis was PD. Temporal bases showed the lowest perfusion; frontal bases and medial parietal lobes the second; visual cortices the third; and parietal association areas exhibited the fourth lowest. During the clinical course, 10 of the patients suffered dementia, 9 had fluctuating cognition, and 19 experienced repeated visual hallucinations. Significant negative correlations were observed between dementia and the bilateral posterior cingulate area, and among fluctuating cognition and bilateral medial parietal lobes, parietal association areas, and dorsal occipital lobes. Repeated visual hallucinations did not show any correlation with any region of interest. We concluded that multiple hypoperfusion areas were observed in the 3D-SSP SPECT analysis. Although the presence of dementia showed a significant relationship with the bilateral posterior cingulate areas, perfusion in the frontal bases, temporal bases, or parietal lobes was markedly more reduced than that seen in the bilateral posterior cingulate areas. PMID- 15806568 TI - Visual system side effects caused by parasympathetic dysfunction after botulinum toxin type B injections. AB - Botulinum toxin type B (BTX-B) has been approved by the Food and Drug Administration for the treatment of cervical dystonia. However, as with botulinum toxin type A (BTX-A) it has off-label uses, such as for hyperhidrosis, focal dystonias, spasticity, and facial wrinkles. BTX-B has also been shown to be a safe and effective alternative for patients who are resistant to BTX-A. The most commonly reported side effects include dry mouth and dysphagia. To date, there have been few reports of visual disturbances associated with BTX-B use. In this study, we report on three individual patients who received BTX-B and who subsequently developed parasympathetic dysfunction of the visual system after injections of BTX-B at remote sites. PMID- 15806569 TI - Properties of titanium-silver alloys for dental application. AB - The purpose of this study was to develop titanium-silver alloys with biocompatibility, high corrosion resistance, and low ion-release rate, and to evaluate the electrochemical properties of titanium-silver alloys in artificial saliva. Titanium-silver alloys with silver contents ranging from 0 to 4.5 at % in steps of 0.5 at % were designed. The alloys were arc melted, homogenized at 950 degrees C for 72 h, hot rolled to 2 mm in thickness, and finally solution heat treated at 950 degrees C for 1 h and quenched in water. Chemical compositions, phases, hardnesses, electrochemical properties, and the cytotoxicity of the alloys were investigated. The purity of titanium-silver alloys was maintained above 99.9%, because few impurities were introduced through their manufacture. In the case of alloys containing silver in the range 2.0-4.0 at %, the formation of an acicular alpha phase was observed inside the beta phase. The acicular phase got thinner with increasing amounts of silver. This means that silver is a beta phase stabilizing element in titanium-silver alloys. The hardness value tended to rise with increasing silver content and increased largely over 3.5 at %, and the increase of the hardness value versus pure titanium was about 33%. It is believed that the substantial increases in hardness was due to the effects of solid solution strengthening and of alpha-beta phase transition. Moreover, titanium silver alloys had higher corrosion resistances than pure titanium. These results mean that silver additions to titanium can improve alloy corrosion resistance. Passive current densities in the potentiodynamic polarization curves were dependent on the chemical compositions of the titanium-silver alloys. However, they did not show a linear relationship with respect to silver contents. Titanium silver alloys did not show pitting corrosion in artificial saliva. It is believed that silver addition to titanium strengthened the passive film due to titanium dissolution induced by the different electromotive forces of titanium and silver. In the agar overlay test, the cytotoxicity of the titanium-silver alloys and of titanium were none or mild. In summary, titanium-silver alloys had higher mechanical properties and corrosion resistance than titanium, and toxicities that were similar to titanium. Therefore, it is recommended that titanium-silver alloys be adopted cautiously by the biomedical and dental fields. PMID- 15806570 TI - Wilson's disease presenting with an unusual cough. AB - A 26-year-old man developed an unusual repetitive, nonproductive cough. Extensive pulmonary and otolaryngology investigations failed to disclose a cause. It was only after he developed additional neurological manifestations ultimately leading to the diagnosis of Wilson's disease (WD) that a neurological basis for the cough was suspected. Features of the cough suggest it was a form of respiratory dyskinesia, a previously unreported presentation of WD. PMID- 15806571 TI - Availability of conjugated ganglioside GD2 monoclonal antibody. PMID- 15806572 TI - Vibrational and electronic circular dichroism of delta-TRISPHAT [tris(tetrachlorobenzenediolato)phosphate(V)] anion. AB - Herein is reported an experimental and theoretical study of the circular dichroism properties of TRISPHAT (1) anion. ECD analysis of the [tetramethylammonium][delta-1] salt confirms the absolute configuration assignment obtained through X-ray crystallographic analysis of the parent cinchonidium salt. The structure, infrared, and vibrational circular dichroism (VCD) spectra derived from density functional theory (DFT) calculations are compared with experimental data. PMID- 15806573 TI - Stereoselective synthesis of an anti-HIV drug candidate. AB - The asymmetric synthesis of a Merck anti-HIV drug candidate is described. The target molecule contains four stereogenic centers, three of which are located in a highly functionalized cyclopentane unit. The convergent synthesis involves the preparation of two key advanced intermediates: the cyclopentane unit and a substituted pyrazole unit. The cyclopentane unit was prepared via two different procedures; a highly diastereoselective Diels-Alder reaction with a chiral oxazolidinone auxiliary and a sequence that incorporated a molybdenum-catalyzed asymmetric allylic alkylation reaction to set the stereocenters. The other key step was a highly diastereoselective hydroxyl-directed reductive amination. The overall yield for the 16-step synthesis was 10%. PMID- 15806574 TI - Characterization of subclinical tremor in Parkinson's disease. AB - The physiological or pathological nature of subclinical tremor amplitude in Parkinson's disease (PD) is not well established. We analyzed characteristics of resting and postural tremors of subclinical amplitude in 17 patients with idiopathic PD without visible resting tremor, having a postural tremor in their least-affected hand rated 0 (12 subjects) or 1 (5 subjects) on Item 21 of the Unified Parkinson's Disease Rating Scale, compared to 17 control subjects matched for age, sex, and handedness. Tremor was recorded at the tip of the index finger using a displacement laser transducer. Overall results show that subclinical resting tremor in PD is significantly different from physiological tremor in terms of amplitude fluctuation, frequency dispersion, harmonic index, and proportional power in 4 to 6 Hz. No significant differences were found for postural tremor. These differences appear to originate mainly from patients with the mixed form of the disease. This study also confirms the preservation of physiological tremor likely originating from a distinct central oscillator in PD. The use of this method in the early and detailed characterization of PD tremors when amplitude is still within normal limits is proposed. PMID- 15806575 TI - Enantiomer separation of a powerful chiral auxiliary, 2-methoxy-2-(1 naphthyl)propionic acid by liquid chromatography using chiral anion exchanger type stationary phases in polar-organic mode; investigation of molecular recognition aspects. AB - The enantiodiscriminating potential of the weak anion exchange-type quinine-based chiral stationary phases (CSPs) for direct enantiomer separation of racemic 2 methoxy-2-(1-naphthyl)propionic acid (selectand, SA) was studied. The influence of structure variations of the selector (SO) in the carbamate functional group and/or in the C6' position of quinoline moiety on retention and enantioselectivity was investigated. Systematic chromatographic studies were made to gain more insight into the overall chiral recognition mechanism for a given mobile phase. In this context, the tert-butylcarbamoyl quinine and the corresponding diisopropylphenyl-derived selector provided the highest resolution and enantioselectivity under polar-organic conditions with the elution order of (R) before the (S) enantiomer. When the bulkiness of the substituents in the C6' position of the SO was increased, the selectivity was decreased in all cases. Alkylation of the nitrogen atom in the carbamate functionality of the SO resulted in the complete loss of enantiomer separation, confirming the crucial importance of the hydrogen-bond formation involved in the stereodiscriminating events. In addition, ten different mono-, bi-, or trivalent acids, necessary as competitor molecules (counter-ions) of the mobile phase, were screened to judge their influence on retention and overall enantioselectivity. Among them, acetic acid, formic acid, N-acetylglycine, and glycolic acid proved to be the most promising counter-ions with R(S) values of 6.35, 6.81, 8.19, and 7.34, respectively. On the basis of chromatographic data, a tentative molecular recognition model was proposed. Simultaneous ion-pairing and hydrogen bonding, in concert with pi-pi stacking and steric interactions, were expected to be responsible for chiral recognition mechanism. This was partially corroborated by structural and/or conformational analysis of the tert-butylcarbamoyl quinine-2-methoxy-2-(1 naphthyl)propionic acid (SO-SA) complex. PMID- 15806576 TI - Neuroanatomy of the normal prostate. AB - BACKGROUND: The prostate is a richly innervated organ, but its neuroanatomy has not been thoroughly mapped and analyzed quantitatively. METHODS: We selected prostates from autopsy cases of cancer-free patients aging from their 40's to 70's, and histologic sections were immunostained with S-100. Nerve density hotspots were mapped, photographed, and analyzed for nerve area. RESULTS: Innervation of the peripheral zone was found to be significantly greater than the transition zone (P < 0.0006), which is in turn significantly more innervated than BPH (P < 0.007). The posterior capsule has significantly more nerve area than the anterior capsule (P < 0.0001). Highest innervation was seen in the neurovascular bundles and seminal vesicles, with the lowest in transition zone and BPH. When compared to increasing patient age, both peripheral zone and overall innervation tended to decrease. CONCLUSIONS: These results are quantitative in nature, performed in cancer-free patients ranging over four different decades of age. We plan to soon compare this profile with our developing profile of cancerous prostates, hoping to learn more about interactions between nerves and prostate cancer. PMID- 15806577 TI - Psychodynamic psychotherapy for separation anxiety disorders in children. PMID- 15806578 TI - Structural features of a central mismatch in oligonucleotide hybrid duplexes visualized via Raman spectroscopy: model system for evaluation of potential "antisense" drugs. AB - Structural features of mismatched base pairs were studied on four nonamer hybrid duplexes formed between the 5'-d(GTGATATGC)-3' complement and its 5'-r(GCAUNUCAC) 3' (N = A, C, G, U) counterparts. This oligonucleotide set is considered a model molecular system for future systematic studies of various modifications of internucleotide linkages with respect to their impact on the structure of mismatched base pairs. Raman spectra, measured at 15 degrees C, revealed the prevailing A-like structure of the RNA strand and mixed A-like and B-like characteristics for the DNA strand. All three mismatches disturb only weakly the overall conformation of the hybrid duplex in contrast to analogous mismatched DNA duplexes. In particular, the dT x rG mismatch influences the global hybrid duplex geometry almost negligibly. The dT x rC and dT x rU mismatches induce somewhat more pronounced distortions of the backbone structure and of the thymine position, the latter being expressed by a change of the surrounding methylene group without effect on the carbonyl's vibrations. Structural effects of the mismatches correlate well with the duplex thermodynamic stabilities obtained by ultraviolet (UV) absorption, i.e., the dT x rG mismatch decreases the hybrid duplex stability very weakly while the effect of both pyrimidine-pyrimidine mismatches is considerable. PMID- 15806579 TI - Response to Dr. Kopke's comments on haplotypes at the OPRM1 locus. PMID- 15806581 TI - Genome-wide linkage analyses of extended Utah pedigrees identifies loci that influence recurrent, early-onset major depression and anxiety disorders. AB - Major depressive disorder (MDD) is a common, clinically heterogeneous disorder often found comorbid with other disorders. We studied recurrent, early-onset MDD (MDD-RE) and anxiety disorders in combination to define powerful phenotypes for genetic study. We used 87 large, extended Utah pedigrees to investigate linkage to 3 phenotypes: "MDD-RE;" "MDD-RE or anxiety;" and "MDD-RE and anxiety;" where in the latter definition the disorders must appear comorbid within an individual. Pedigrees ranged in size from 2 to 6 generations and contained 3 to 42 individuals affected with MDD or anxiety (718 total). In primary analyses, we identified three regions with at least suggestive genome-wide evidence for linkage on chromosomes 3centr, 7p, and 18q. Both 7p and 18q are replication findings for related phenotypes. The best linkage evidence was for a novel locus at 3p12.3-q12.3 (LOD = 3.88, "MDD-RE or anxiety") and 18q21.33-q22.2 (LOD = 3.75, "MDD-RE and anxiety"), a well-established susceptibility locus for bipolar disorder. In our secondary sex-specific analyses, we identified two further regions of interest on chromosomes 4q and 15q. Using linked pedigrees, we localized 3centr and 18q to 9.8 and 12.2 cM, respectively, with potential for further localization with the addition of markers in specific pedigrees. Our success in replication and novel locus identification illustrates the utility of large extended pedigrees for common disorders, such as MDD. Further, it supports the hypothesis that MDD and anxiety disorders have over-lapping genetic etiologies and suggests that comorbid diagnoses may be useful in defining more genetically homogeneous forms of MDD for linkage mapping. PMID- 15806582 TI - Mutational screening and association study of glutamate decarboxylase 1 as a candidate susceptibility gene for bipolar affective disorder and schizophrenia. AB - Recent evidence from postmortem studies suggests that GAD1 encoding the gamma aminobutyric acid (GABA) synthetic enzyme GAD67 is a functional candidate susceptibility gene for both bipolar affective disorder (BPAD) and schizophrenia. Previous studies suggest linkage between D2S326 near GAD1 and BPAD. We systematically screened GAD1 exons, flanking intronic sequences, and the promoter sequence for polymorphisms in 16 BPAD patients and five controls from Denmark. We identified eight single nucleotide polymorphisms (SNPs) including two in the promoter sequence. An association study of SNPs covering GAD1 was performed in a Danish sample of 82 BPAD subjects and 120 controls and in a Scottish sample of 197 individuals with schizophrenia, 200 BPAD subjects and 199 controls. Linkage disequilibrium (LD) and haplotype frequencies were estimated from genotype data from eight SNPs. Strong pairwise LD was observed among all pairs of neighboring markers. In the Danish sample, we found weak association between BPAD and two promoter SNPs spaced 1 kb apart. Furthermore, one, two, and three loci haplotype analysis showed weak association with BPAD in the Danish sample. The results from the association studies indicate that promoter variants are of importance for the Danish BPAD cases and we cannot reject the hypothesis of GAD1 as a functional candidate gene for BPAD. No association was observed between BPAD or schizophrenia and any of the investigated SNPs in the Scottish sample set. Thus the results obtained from the Scottish sample suggest that the GAD1 gene variants do not play a major role in the predisposition to schizophrenia. PMID- 15806583 TI - Association of the serotonin transporter promoter polymorphism with smoking behavior among adolescents. AB - Serotonin transporter promoter polymorphism (5-HTTLPR) genotype was previously found associated with smoking behavior, difficulty in quitting smoking, and nicotine addiction; with non-replicated findings and contrasting results. Aim of the present study was to evaluate the possible association between 5-HTTLPR genotype and smoking behavior among adolescents, in relationship with psychological characteristics. Two hundred and ten Caucasian high school students (aged 14-19 years); 103 non-smokers, who have never smoked nicotine; and 107 tobacco smokers have been genotyped. Aggressiveness levels and temperamental traits were measured in both smokers and non-smokers, respectively, utilizing Buss-Durkee Hostility Inventory (BDHI) and Cloninger Three-Dimensional Personality Questionnaire (TPQ). Data about school performance have been also collected. The short-short (SS) genotype frequency was significantly higher among smokers compared with non-smokers (P = 0.023). The odds ratio for the SS genotype versus the long-long (LL) genotype frequency was 1.17 [95% CL (0.30-2.05)], when smokers were compared with non-smokers. The SS genotype frequency was significantly higher among heavy smokers with early onset, compared with moderate smokers with late onset (P = 0.042). BDHI irritability scores, NS scores at TPQ, and school failure frequency were significantly higher in smokers than in non smokers. Multivariate model-fitting analysis evidenced a significantly greater relationship of genotype with irritability levels (BDHI scores) (0.34, P < 0.001) and temperament traits (NS scores) (0.36, P < 0.001), than with school performance (rate of school under-achievements) (0.18, P < 0.05) and nicotine smoking (number of cigarettes) (0.24, P < 0.01). Accordingly, factor-analysis showed that gene polymorphism contributes more directly to BDHI scores and NS scores (0.73; 0.71) than to smoking behavior and school under-achievement (0.54; 0.51). Our data suggest that a decreased expression of the gene encoding the 5 HTT transporter, due to "S" promoter polymorphism, may be associated with smoking behavior among adolescents and increased risk to develop nicotine dependence, possibly in relationship to personality traits, temperamental characteristics, and school under-achievements. PMID- 15806584 TI - Toward localizing genes underlying cerebral asymmetry and mental health. AB - Genome investigations of autism, attention deficit hyperactivity disorder (ADHD), and dyslexia suggest possible genetic overlap. Atypical cerebral asymmetry (ACA), the absence of the left hemisphere dominance for language, may be a shared phenotype due to genes located in regions of overlap. A binomal test is used to evaluate whether linked regions overlap more than expected by chance for 15 genome-wide scans in autism, ADHD, and dyslexia. Significant evidence of linkage overlap (P = 10(-7)) is seen for autism, ADHD, and dyslexia for seven chromosomal regions (2p11-12, 5p13, 7q22-33, 9q33-34, 13q22, 16p13, and 17p11-q11). Linkage analysis of ACA and molecular markers for 270 sibling pairs with ADHD is conducted using the Haseman-Elston statistic. Linkage analysis supports ACA as a shared phenotype with risk genes located on 9q33-34 or 16p13 (P < 0.004). Further support stems from the overlap of these regions in schizophrenia, bipolar illness, specific language impairment (SLI), and handedness, all traits associated with ACA. Autism, ADHD, and dyslexia share regions of linkage overlap and ACA may be a shared phenotype for such genes similar to HLA in autoimmune disease. Because ACA is associated with certain aspects of creativity, such risk genes may also be enhancer genes for creativity. PMID- 15806585 TI - Validated toxicological determination of 30 drugs of abuse as optimized derivatives in oral fluid by long column fast gas chromatography/electron impact mass spectrometry. AB - An analytical procedure was developed for the simultaneous sensitive identification, screening and quantitation of 30 drugs of abuse using 250 microl of human oral fluid. The method employs sequential mixed-mode solid-phase extraction (SPE), optimized derivative formation and long-column fast gas chromatography/electron impact mass spectrometry (GC/EI-MS). After sequential SPE elution, the most sensitive and stable derivatives were formed by taking careful account of the characteristics of the active functional groups and possible steric hindrances affecting derivatization chemistry. Amphetamine-type stimulant drugs were acylated with heptafluorobutyric anhydride, benzodiazepines and Delta(9)-tetrahydrocannabinol were silylated with N-methyl-N-(tert butyldimethylsilyl)trifluoroacetamide and benzoylecgonine, codeine, ethylmorphine, 6-monoacetylmorphine, morphine, pholcodine, buprenorphine and norbuprenorphine with N-methyl-N-(trimethylsilyl)trifluoroacetamide. In addition, the following analytes were included: methadone, cocaine, alprazolam, midazolam, fentanyl and zolpidem. In GC separation, fast temperature ramping and high carrier gas flow-rate combined with long 30 m columns of i.d. 0.32 mm offered a reduction in analysis time and sharp peak shapes while still maintaining sufficient resolution and high sample capacity. Validated parameters including selectivity, linearity, accuracy, intra- and inter-day precision, extraction efficiency and limit of quantitation were all within required limits. In contrast to previously published methods, this single procedure is suitable for the simultaneous toxicological determination of the most common illicit drugs and benzodiazepines, and also zolpidem, in a small amount of oral fluid. PMID- 15806586 TI - Embryology of neural tube development. AB - Neurulation is the process of forming the neural tube, which will become the brain and spinal cord. This article reviews the various cellular processes involved in neurulation and discusses possible roles of folate in this process. PMID- 15806587 TI - The sensitivity of antenatal ultrasound for predicting renal tract surgery in early childhood. AB - OBJECTIVE: To establish the sensitivity of antenatal ultrasound for identifying the need for renal tract surgery in infancy and early childhood. METHODS: A retrospective analysis of the surgical records in children under 5 years of age undergoing renal tract surgery in a regional pediatric urological surgery referral unit was carried out. All records between May 1997 and July 2002 were examined to assess the relationship between prenatal ultrasound findings and postnatal surgical pathology. RESULTS: A total of 106 operations had been performed. The detection rate of multicystic renal dysplasia was 100% (17/17). The equivalent detection rates for pelviureteral junction obstruction, duplex renal system and vesicoureteral reflux were 82.8, 67 and 26.1%, respectively. None of the babies with renal tumors had abnormal antenatal ultrasound findings. The diagnosis of postnatal renal surgical pathology was made on the basis of prenatal scan findings in 59.6% of cases, while recurrent urinary tract infection led to the diagnosis in 26.0%. CONCLUSIONS: Approximately 40% of children requiring surgery for renal tract pathology will have a normal antenatal ultrasound examination. The prevalence of abnormal antenatal ultrasound findings varies depending on the type of renal tract pathology. Despite these findings, the commonest indicator for surgery remains abnormal prenatal scan findings, followed by recurrent urinary tract infections. PMID- 15806589 TI - Huntington's disease and minocycline. PMID- 15806591 TI - Re: "Two new radiological findings to improve the diagnosis of bronchial foreign body aspiration in children" by Girardi et al. (Pediatr Pulmonol 2004;38:261 264). PMID- 15806594 TI - Neural tube closure and neural tube defects: studies in animal models reveal known knowns and known unknowns. AB - The vertebrate central nervous system is a hollow structure that develops first as a flat sheet of cells and subsequently rolls into a tube during embryogenesis. Failure of this rolling process, called neural tube closure, results in a class of common human birth defects called neural tube defects. The cellular and molecular mechanisms governing neural tube closure have been studied extensively in animal models, but much remains to be elucidated. In this review, I will highlight recent progress in understanding neural tube closure mechanisms and how these studies can inform our search for the genes that underlie human neural tube defects. Supplementary material for this article can be found on the American Journal of Medical Genetics (Part C) website (http://www.mrw.interscience.wiley.com/suppmat/1552 4868/suppmat/2005/135/v135.1.wallingford.html) PMID- 15806593 TI - The CFTR gene and regulation of its expression. AB - The cystic fibrosis transmembrane conductance regulator gene (CFTR) shows clear temporal and developmental regulation of its expression. However, there are few well-defined regulatory elements that control this pattern of expression, and their mechanism of action is poorly understood. We review the structure and organization of the CFTR gene and what is known about its regulation. The CFTR gene promoter is clearly important for maintaining levels of CFTR gene expression, but apparently it does not contain any tissue-specific elements. Thus tissue-specificity is probably controlled by sequences lying elsewhere in this large gene. We discuss data from our group and others implicating additional regions of CFTR in regulatory functions, and evaluate candidate transcription factors that may be involved. Further, we summarize aspects of the regulation of the developmental expression of CFTR. Definition of CFTR gene regulatory elements could be of considerable therapeutic significance, since only a small increase in CFTR expression in the correct cell type could alleviate the disease phenotype. PMID- 15806595 TI - Beta 2 adrenergic receptor polymorphisms in cystic fibrosis. AB - There has been a recent emphasis on identifying modifier genes that influence the severity of cystic fibrosis (CF) lung disease. The beta-2-adrenergic receptor is expressed on airway smooth muscle, is the target for inhaled beta agonists, and has several common polymorphisms in its gene, ADRB2. Polymorphisms changing glycine to arginine or glutamate to glutamine in codons 16 and 27, respectively, were associated with differences in clinical response to inhaled beta agonists in individuals with asthma. We compared acute airway responsiveness and 5-year decline in pulmonary function in CF patients with different ADRB2 genotypes. One hundred and six subjects performed spirometry before and after the administration of an inhaled bronchodilator, and had ADRB2 genotype determined for codons 16 and 27. Comparing the percent change in FEV(1) and FEF(25-75) continuously revealed differences in the degree of airway responsiveness to bronchodilator between ADRB2-genotyped groups. However, there was no significant relationship between the ADRB2 genotype at positions 16 and 27 and bronchodilator response when defined as 12% improvement in FEV(1). Five-year decline in percent predicted FEV(1) showed no association with ADRB2 genotype. These data are consistent with variants of the ADRB2 gene having different responses to bronchodilator, but the long-term effects, if any, are not apparent over a 5-year period. PMID- 15806596 TI - Correlation between cough frequency and airway inflammation in children with primary ciliary dyskinesia. AB - Cough is common in airway disease. We measured cough frequency in children with primary ciliary dyskinesia (PCD), to determine how accurately families assess this symptom; and to assess the relationship between cough frequency and airway inflammation, measured using induced sputum and exhaled nitric oxide (eNO). Twenty stable PCD children (7 boys), median age 10.8 years (interquartile range (IQR), 9-14), and 10 healthy control children, median age 12 years (IQR, 10.5 12.7), were recruited. ENO was measured using a chemiluminescence analyzer, with sputum induction with 3.5% saline. PCD children underwent ambulatory cough monitoring. Sputum neutrophils were higher in PCD (median, 70.3%; IQR, 55.3-78%) compared to controls (median, 27%; IQR, 24.5-33%; P = 0.004); cough frequency was higher (median episodes, 19; IQR, 11-22.5) compared to healthy children (median episodes, 6.7; IQR, 4.1-10.5; P < 0.001). Forced expiratory volume in 1 sec (FEV(1) percent predicted) and eNO were lower in PCD (median, 63%; IQR, 57-85%; P < 0.0001); eNO (median, 7.1 ppb (IQR, 4.8-19.1 ppb) vs. 12.4 ppb (IQR, 10.3-17.3 ppb), P = 0.043). Parental scoring of day and night cough correlated with recorded cough (r = 0.930, P < 0.0001, daytime; r = 0.711 for nighttime, P = 0.002). Visual analogue score and cough episodes also correlated positively (r = 0.906; P < 0.0001). There was a positive correlation between cough frequency and sputum neutrophil count in PCD (Spearman's r = 0.693, P < 0.002), but not percent FEV(1) or eNO. Stable PCD children have increased cough frequency and neutrophilic airway inflammation. In conclusion, cough frequency correlated with sputum neutrophils but not with FEV1 or eNO. PMID- 15806597 TI - Standard criteria for relapse are needed in obsessive-compulsive disorder. AB - To assess how different criteria for relapse affect inferences about relapse in obsessive-compulsive disorder (OCD), a post hoc analysis of relapse was conducted using data from a multisite randomized controlled trial comparing clomipramine (CMI), exposure and ritual prevention (EX/RP), and its combination (EX/RP+CMI) in adults with OCD. Different relapse definitions were constructed based on criteria used in prior studies. For each definition, the number of relapsers was computed, and the proportion of relapsers and time to relapse were compared. When applied to this data set, relapse criteria used in prior OCD studies yielded different observed relapse rates (range: 27-63% for CMI; 0-50% for EX/RP; and 7-67% for EX/RP+CMI). Most criteria found that EX/RP responders (with or without CMI) had a significantly lower relapse rate and longer time to relapse after treatment discontinuation than did responders to CMI alone. However, some relapse criteria (e.g., those requiring minimal worsening) found no significant treatment differences in relapse rates or time to relapse, and some generated biases against one treatment or another. Most definitions concurred: in adults with primary OCD, EX/RP treatment (with or without CMI) can produce more durable short term gains after treatment discontinuation than CMI alone. However, different relapse criteria can lead to very different observed relapse rates and even contradictory inferences about relapse. Standard criteria for relapse are needed in OCD to facilitate comparisons between studies (enabling better treatment guidelines) and to advance research on mechanisms of relapse and relapse prevention. PMID- 15806598 TI - Association of the apolipoprotein A-IV: 360 Gln/His polymorphism with cerebrovascular disease, obesity, and depression in a Brazilian elderly population. AB - The identification of genetic polymorphisms as risk factors for complex diseases can be relevant for their prevention, diagnosis, and prognosis. The apolipoprotein A-IV: 360 Gln/His polymorphism was investigated in 383 elderly individuals, who were participants of a longitudinal study commenced in 1991. The major morbidities that affect elderly people, such as cardiovascular diseases, diabetes, low cognitive function, depression, and obesity, were extensively investigated. DNA was isolated from blood cells, amplified by PCR, and digested with Fnu4HI. In this population the frequency of the His allele was 0.056 and the genotypes were distributed according to Hardy-Weinberg equilibrium. Logistic regression analysis showed a significant association between the presence of His allele and cerebrovascular disease and/or transitory ischemic attack (odds ratio) (OR = 3.070, P = 0.027), obesity (OR = 2.241, P = 0.047), and depression (OR = 2.879, P = 0.005). This study indicates that the presence of the rare allele in elderly people can play a significant role in the occurrence of multifactorial diseases. This is the first study analyzing this polymorphism in elderly people in Brazil. More studies should be encouraged to elucidate the mechanisms involved in these diseases. PMID- 15806599 TI - Differential expression of matrix metalloproteinases and interleukin-8 during regeneration of human airway epithelium in vivo. AB - In many airway diseases, the airway epithelium is severely damaged and has to regenerate rapidly to restore its function. The regeneration process involves chronological steps of epithelial cell migration, proliferation, stratification, and differentiation. The present study has used an in vivo humanized airway xenograft model in nude mice that mimics the regeneration dynamics of human airway epithelium after severe injury, and human-specific molecular tools, to study the expression profiles of epithelial matrix metalloproteinases (MMPs)-7 and -9, of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), and of the pro-inflammatory cytokine interleukin-8 (IL-8) during the different steps of human airway epithelium regeneration. It was found that during the cell migration and proliferation steps, airway epithelial cells expressed IL-8 at a high level, whereas airway epithelial pseudo-stratification and surface airway epithelial differentiation were associated with increased expression of MMPs and a progressive decrease in IL-8. Interestingly, immunohistochemical analysis revealed exclusive expression of MMPs at the apical part of the well differentiated regenerated airway epithelium, and incubation of the regenerating epithelial cells with MMP inhibitors led to abnormal epithelial differentiation. These data provide new insight into the temporal expression of MMPs and IL-8 during the regeneration of airway epithelium and demonstrate the involvement of these factors during the different steps that lead to restoration of a well differentiated and functional airway epithelium. PMID- 15806600 TI - Stability of (C60)2 and epoxide dimers, (C60)2ON, and their anions. AB - The PM3, AM1, and MINDO3 semiemperical methods are used to calculate the the energy difference between C60ON and C60ON- and the bond dissociation energy necessary to cleave neutral and negatively charged (C60)2 dimers and epoxide dimers, (C60)2ON, to their respective monomers C60, and C60ON/2. The results show that the anions of the dimers are significantly more stable than neutral dimers. This result may explain the higher thermal stability of the observed ferromagnetic phase in photolyzed C60. which has been attributed to epoxide dimers and oligomers. It also provides an explanation for the origin of unpaired electron spin necessary for ferromagnetism. PMID- 15806601 TI - Monoamine oxidase A (MAOA) and antisocial behaviors in the presence of childhood and adolescent maltreatment. AB - There is a robust relationship between the experience of maltreatment in childhood and later antisocial behaviors amongst adolescents and adults. Animal and human studies suggest that variation in monoamine oxidase A (MAOA) genotype may moderate the effects of maltreatment. Self-reported conduct problems and criminal convictions amongst sibling-pairs from the National Longitudinal Study of Adolescent Health were tested for association with reports of maltreatment before and after the age of 12. MAOA promoter polymorphisms were tested for possible moderation effects. Maltreatment predicted conduct problems and criminal convictions. MAOA genotype did not have a significant moderating effect in any of the six analyses that were conducted. We did not replicate a previous report that MAOA polymorphisms moderated the relationship between maltreatment and conduct problems. There was, however, a non-significant trend in the predicted direction. Additional studies will be needed before firm conclusions can be drawn about this hypothesized genotype-environment interaction. PMID- 15806602 TI - Molecular dynamics simulation of clustered DNA damage sites containing 8 oxoguanine and abasic site. AB - Clustered DNA damage sites induced by ionizing radiation have been suggested to have serious consequences to organisms, such as cancer, due to their reduced probability to be repaired by the enzymatic repair machinery of the cell. Although experimental results have revealed that clustered DNA damage sites effectively retard the efficient function of repair enzymes, it remains unclear as to what particular factors influence this retardation. In this study, approaches based on molecular dynamics (MD) simulation have been applied to examine conformational changes and energetic properties of DNA molecules containing clustered damage sites consisting of two lesioned sites, namely 7,8 dihydro-8-oxoguanine (8-oxoG) and apurinic/apyrimidinic (AP) site, located within a few base pairs of each other. After 1 ns of MD simulation, one of the six DNA molecules containing a clustered damage site develops specific characteristic features: sharp bending at the lesioned site and weakening or complete loss of electrostatic interaction energy between 8-oxoG and bases located on the complementary strand. From these results it is suggested that these changes would make it difficult for the repair enzyme to bind to the lesions within the clustered damage site and thereby result in a reduction of its repair capacity. PMID- 15806603 TI - Carbon boronyls: species with higher viable possibility than boron carbonyls at the density functional theory. AB - Density functional theory investigations indicate that carbon boronyls (CBO)n (n = 3-7) are considerably more stable in thermodynamics than their boron carbonyl isomers (BCO)n and exhibit aromaticity throughout the whole series. The extra stabilities of (CBO)n originate from their frontier pi molecular orbitals delocalized over the Dnh Cn central rings which are absent in (BCO)n. It is expected that experimental characterization of these (CBO)n species may open a new branch of chemistry on carbon boronyls. PMID- 15806604 TI - Quantitative analysis of wine yeast gene expression profiles under winemaking conditions. AB - Wine fermentation is a dynamic and complex process in which the yeast cell is subjected to multiple stress conditions. A successful adaptation involves changes in gene expression profiles where a large number of genes are up- or downregulated. Functional genomic approaches are commonly used to obtain global gene expression profiles, thereby providing a comprehensive view of yeast physiology. We used SAGE to quantify gene expression profiles in an industrial strain of Saccharomyces cerevisiae under winemaking conditions. The transcriptome of wine yeast was analysed at three stages during the fermentation process, mid exponential phase, and early- and late-stationary phases. Upon correlation with the yeast genome, we found three classes of transcripts: (a) sequences that corresponded to ORFs; (b) expressed sequences from intergenic regions; and (c) messengers that did not match the published reference yeast genome. In all fermentation phases studied, the most highly expressed genes related to energy production and stress response. For many pathways, including glycolysis, different transcript levels were observed during each phase. Different isoenzymes, including hexose transporters (HXT), were differentially induced, depending on the growth phase. About 10% of transcripts matched non-annotated ORF regions within the yeast genome and could correspond to small novel genes originally omitted in the first gene annotation effort. Up to 22% of transcripts, particularly at late-stationary phase, did not match any known location within the genome. As the available reference yeast genome was obtained from a laboratory strain, these expressed sequences could represent genes only expressed by an industrial yeast strain. Further studies are necessary to identify the role of these potential genes during wine fermentation. PMID- 15806605 TI - Hyperhomocysteinemia, methylenetetrahydrofolate reductase 677TT genotype, and the risk for schizophrenia: a Dutch population based case-control study. AB - Evidence for an involvement of aberrant homocysteine metabolism in the aetiology of schizophrenia is limited and controversial. A case-control study was performed to quantify the risk of schizophrenia in the presence of elevated homocysteine concentrations or homozygosity for the 677C --> T polymorphism (677TT) in the methylenetetrahydrofolate reductase (MTHFR) gene in subjects of Dutch ancestry. We determined the 677C --> T MTHFR genotype distribution in 254 well-defined patients and 414 healthy controls. Plasma homocysteine concentrations were measured in 62 patients with schizophrenia and 432 control subjects. When homocysteine concentrations were stratified into quartiles of the control distribution, we calculated an increased risk for schizophrenia in the fourth and third quartile versus the lowest quartile [odds ratio (OR) = 3.3; 95% confidence interval (CI): 1.2-9.2, and OR = 3.1; 95% CI: 1.2-8.0, respectively]. A significant dose-response relation of increasing homocysteine levels and increasing risk for schizophrenia was observed (P = 0.036). The 677TT genotype was associated with an OR of 1.6 [95% CI: 0.96-2.8] of having schizophrenia. Heterozygosity for the T allele compared to 677CC subjects accounted for an OR of 1.3 [95% CI: 0.91-1.8]. Elevated homocysteine levels and the MTHFR 677TT genotype are associated with an increased risk for schizophrenia. These observations support a causal relation between disturbed homocysteine metabolism and schizophrenia. PMID- 15806606 TI - Strong means slow: dynamic contributions to the bulk mechanical properties of supramolecular networks. PMID- 15806607 TI - Total synthesis of antascomicin B. PMID- 15806608 TI - Gold catalysis: proof of arene oxides as intermediates in the phenol synthesis. PMID- 15806609 TI - Glory-scattering measurement of water-noble-gas interactions: the birth of the hydrogen bond. PMID- 15806610 TI - SFP1 is involved in cell size modulation in respiro-fermentative growth conditions. AB - Saccharomyces cerevisiae grows fast on glucose, while growth slows down on ethanol as cells move from glucose fermentation to oxidative metabolism. The type of carbon source influences both the specific growth rate and cell cycle progression, as well as cell size. Yeast cells grown on glucose have a larger size than cells grown on ethanol. Here, we analysed the behaviour of a sfp1 null mutant during balanced and transitory states of growth in batch in response to changes in the growth medium carbon sources. In a screening for mutants affected in cell size at Start, SFP1 has been identified as a gene whose deletion caused one of the smallest whi phenotype. Findings presented in this work indicate that in the sfp1 null mutant the reduction in cell size is not only a consequence of the reduced growth rate but it is tightly linked to the cellular metabolism. The SFP1 gene product is required to sustain the increase of both rRNA and protein content that in wild-type cells takes place in respiro-fermentative growth conditions, while it seems dispensable for growth on non-fermentable carbon sources. It follows that sfp1 cells growing on ethanol have a larger size than cells growing on glucose and, noticeably, the former enter the S phase with a critical cell size higher than the latter. These features, combined with the role of Sfp1p as a transcriptional factor, suggest that Sfp1p could be an important element in the control of the cell size modulated by nutrients. PMID- 15806611 TI - Self-assembly and cross-linking of bionanoparticles at liquid-liquid interfaces. PMID- 15806612 TI - In vivo specificity of Ure2 protection from heavy metal ion and oxidative cellular damage in Saccharomyces cerevisiae. AB - The S. cerevisiae Ure2 protein is a prion precursor able to form large homopolymers with the characteristics of amyloid particles, a function largely restricted to its 90 N-terminal amino acids. The remaining C-terminal domain of Ure2 plays two important roles in cellular metabolism. First, it regulates nitrogen catabolic gene expression by forming a complex with the GATA transcription factor Gln3. This complex formation correlates with Gln3 being sequestered in the cytoplasm under conditions of excess nitrogen, where Gln3/Gat1 mediated transcription is minimal. Second, Ure2, which possesses structural homology with glutathione S-transferases and binds to xenobiotics and glutathione, has been recently shown to be required for Cd(II) and hydrogen peroxide detoxification. Present experiments demonstrate that Ure2 possesses a far broader protection specificity, being required to avoid the toxic effects of As(III), As(V), Cr(III), Cr(VI), Se(IV), as well as Cd(II) and Ni(II), and to varying lesser degrees Co(II), Cu(II), Fe(II), Ag(I), Hg(II), cumene and t-butyl hydroperoxides. In contrast, deletion of URE2 greatly enhances a cell's ability to withstand toxic concentrations of Zn(II) and Mo(VI). In the case of Cd(II), Ure2 does not function to decrease intracellular Cd(II) levels or influence glutathione availability for glutathionation. In fact, ure2 hypersensitivity to Cd(II) remains the same, even when glutathione is used as sole source of nitrogen for cell growth. These data suggest that Ure2 possesses a central role in metal ion detoxification, a role not demonstrably shared by either of the two known S. cerevisiae glutathione S-transferases, Gtt1 and Gtt2, or the two glutaredoxins, Grx1 and Grx2, that also possess glutathione S-transferase activity. PMID- 15806613 TI - Investigation of limiting metabolic steps in the utilization of xylose by recombinant Saccharomyces cerevisiae using metabolic engineering. AB - A Saccharomyces cerevisiae screening strain was designed by combining multiple genetic modifications known to improve xylose utilization with the primary objective of enhancing xylose growth and fermentation in xylose isomerase (XI) expressing strains. Strain TMB 3045 was obtained by expressing the XI gene from Thermus thermophilus in a strain in which the GRE3 gene coding for aldose reductase was deleted, and the genes encoding xylulokinase (XK) and the enzymes of the non-oxidative pentose phosphate pathway (PPP) [transaldolase (TAL), transketolase (TKL), ribose 5-phosphate ketol-isomerase (RKI) and ribulose 5 phosphate epimerase (RPE)] were overexpressed. A xylose-growing and fermenting strain (TMB 3050) was derived from TMB 3045 by repeated cultivation on xylose medium. Despite its low XI activity, TMB 3050 was capable of aerobic xylose growth and anaerobic ethanol production at 30 degrees C. The aerobic xylose growth rate reached 0.17 l/h when XI was replaced with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes expressed from a multicopy plasmid, demonstrating that the screening system was functional. Xylose growth had not previously been detected in strains in which the PPP genes were not overexpressed or when overexpressing the PPP genes but having XR and XDH genes chromosomally integrated. This demonstrates the necessity to simultaneously increase the conversion of xylose to xylulose and the metabolic steps downstream of xylulose for efficient xylose utilization in S. cerevisiae. PMID- 15806614 TI - A systematic nomenclature of chromosomal elements for hemiascomycete yeasts. AB - We present a compact, stable, unambiguous and extensible nomenclature for unique chromosomal elements from genomic DNA, developed to meet the increasing need created by the increasing number of yeast sequencing projects. Our proposal, adopted for use in the Genolevures project, is specifically designed to facilitate basic tasks in comparative genomics. PMID- 15806615 TI - Rapid enrichment and analysis of yeast phosphoproteins using affinity chromatography, 2D-PAGE and peptide mass fingerprinting. AB - A combination of affinity purification, 2D-PAGE and peptide mass fingerprinting was employed to study the phosphoprotein complement of Saccharomyces cerevisiae. Protein extracts were first passed through a phosphoprotein affinity column, and the phosphoprotein-enriched eluate fractions were then separated on 2D gels and visualized by staining with SYPRO Ruby. Proteins were excised from the gels and identified by peptide mass fingerprinting; 11/13 protein spots identified from a gel of the phosphoprotein-enriched fraction had prior published evidence indicating that they were phosphoproteins. Additional experiments using a specific stain for phosphoproteins, prior incubation of the protein extract with alkaline phosphatase and blotting with monoclonal antibodies to phosphothreonine, phosphoserine and phosphotyrosine demonstrated that the phosphoprotein affinity column was an effective method for enriching phosphoproteins. Further validating the method, growth of yeast in the presence of sorbic acid resulted in altered phosphorylation of 17 proteins, 13 of which had prior published evidence that they were phosphoproteins or had ATP binding activity. PMID- 15806616 TI - Bias of estimates of the number needed to treat. AB - There are several commonly used measures of association between treatment and control event rates in the population (piT and piC, respectively). One such measure, the number needed to treat (NNT) indicates the number of patients, on average, who must be treated in order to prevent one additional adverse event, and is equal to 1/(piC - piT). Because the population values piC and piT are unknown, the sample proportions (rates) pC and pT are used as estimates. The precision of a sample-based estimator is usually exhibited in terms of confidence intervals. However, the accuracy of the estimator (i.e., its bias) is often ignored. The purpose of the present study is to examine the degree of bias. Using exact calculations based on the binomial theorem, we determined the bias of an estimate of NNT conditional on pC not equal pT, and the bias of an adjusted estimator of the NNT for various sample sizes (n= 10, 20, 30, 40, 50, 100) and population parameters (0.01 < or = piC < or = 0.9; 0.01 < or = piC - piT < or = 0.8). The magnitude and non-monotonic nature of the bias are due to the NNT scale. The bias of the adjusted estimator can be approximated for some studies using the tabular results in this analysis. PMID- 15806617 TI - Gene selection for microarray data analysis using principal component analysis. AB - Principal component analysis (PCA) has been widely used in multivariate data analysis to reduce the dimensionality of the data in order to simplify subsequent analysis and allow for summarization of the data in a parsimonious manner. It has become a useful tool in microarray data analysis. For a typical microarray data set, it is often difficult to compare the overall gene expression difference between observations from different groups or conduct the classification based on a very large number of genes. In this paper, we propose a gene selection method based on the strategy proposed by Krzanowski. We demonstrate the effectiveness of this procedure using a cancer gene expression data set and compare it with several other gene selection strategies. It turns out that the proposed method selects the best gene subset for preserving the original data structure. PMID- 15806618 TI - Evaluating survival model performance: a graphical approach. AB - In the last decade, many statistics have been suggested to evaluate the performance of survival models. These statistics evaluate the overall performance of a model ignoring possible variability in performance over time. Using an extension of measures used in binary regression, we propose a graphical method to depict the performance of a survival model over time. The method provides estimates of performance at specific time points and can be used as an informal test for detecting time varying effects of covariates in the Cox model framework. The method is illustrated on real and simulated data using Cox proportional hazard model and rank statistics. PMID- 15806619 TI - The value of information and optimal clinical trial design. AB - Traditional sample size calculations for randomized clinical trials depend on somewhat arbitrarily chosen factors, such as type I and II errors. Type I error, the probability of rejecting the null hypothesis of no difference when it is true, is most often set to 0.05, regardless of the cost of such an error. In addition, the traditional use of 0.2 for the type II error means that the money and effort spent on the trial will be wasted 20 per cent of the time, even when the true treatment difference is equal to the smallest clinically important one and, again, will not reflect the cost of making such an error. An effectiveness trial (otherwise known as a pragmatic trial or management trial) is essentially an effort to inform decision-making, i.e. should treatment be adopted over standard? As such, a decision theoretic approach will lead to an optimal sample size determination. Using incremental net benefit and the theory of the expected value of information, and taking a societal perspective, it is shown how to determine the sample size that maximizes the difference between the cost of doing the trial and the value of the information gained from the results. The methods are illustrated using examples from oncology and obstetrics. PMID- 15806620 TI - Separation and aquatic toxicity of enantiomers of synthetic pyrethroid insecticides. AB - Synthetic pyrethroid insecticides are chiral compounds with multiple asymmetric positions. In this study, isomers of four commonly used pyrethroids were separated at the enantiomeric level by enantioselective high-performance liquid chromatography (HPLC), and differences between enantiomers in aquatic toxicity were characterized using individual isomers. Isomers of cis-bifenthrin and permethrin were completely resolved on a Sumichiral OA-2500-I column. All eight isomers of cypermethrin and cyfluthrin were completely separated on two chained Chirex 00G-3019-DO columns. Great differences were found between enantiomers in the acute toxicity to aquatic invertebrates Ceriodaphnia dubia or Daphnia magna. In cis-bifenthrin (cis-BF) and cis-permethrin (cis-PM), the 1R-cis isomer was 15 38 times more active than the 1S-cis enantiomer, while in trans-PM, the 1R-trans isomer was substantially more toxic than the 1S-trans enantiomer. In cypermethrin or cyfluthrin, two of the eight isomers, 1R-cis-alphaS and 1R-trans-alphaS, contributed for almost all the toxicity in the racemate, while the other six enantiomers were inactive. These results suggest that significant enantioselectivity occurs for pyrethroids in aquatic toxicity, and such enantiomeric differences must be considered when evaluating ecological effects of pyrethroid insecticides. PMID- 15806621 TI - Covariate-adjusted adaptive randomization in a sarcoma trial with multi-stage treatments. AB - We present a Bayesian design for a multi-centre, randomized clinical trial of two chemotherapy regimens for advanced or metastatic unresectable soft tissue sarcoma. After randomization, each patient receives up to four stages of chemotherapy, with the patient's disease evaluated after each stage and categorized on a trinary scale of severity. Therapy is continued to the next stage if the patient's disease is stable, and is discontinued if either tumour response or treatment failure is observed. We assume a probability model that accounts for baseline covariates and the multi-stage treatment and disease evaluation structure. The design uses covariate-adjusted adaptive randomization based on a score that combines the patient's probabilities of overall treatment success or failure. The adaptive randomization procedure generalizes the method proposed by Thompson (1933) for two binomial distributions with beta priors. A simulation study of the design in the context of the sarcoma trial is presented. PMID- 15806622 TI - [Reports from the annual meeting of the American Thoracic Society, Orlando, Florida, USA, 21-26 May 2004]. PMID- 15806684 TI - 2005 resource guide. PMID- 15806683 TI - Relationship between stroke volume and sympathetic nerve activity: new insights about autonomic mechanisms of syncope. AB - Head-up tilt table experiments conducted in astronauts prior to and immediately after the NASA Neurolab Space Mission (STS-90) revealed that a reduction in stroke volume induced by moving from the supine to upright posture was associated with increased muscle sympathetic nerve activity (MSNA). Although this finding was not unexpected, lower average stroke volume and greater average MSNA measured after space flight in both supine and upright postures were positioned in a linear fashion on the same stroke volume-MSNA stimulus-response relationship as the average pre-flight stroke volume and MSNA responses. Since all astronauts who participated in the Neurolab orthostatic experiments completed the 10-min tilt table tests, these observations supported the notion that sympathetic reflex responses were not altered but functioned adequately after space flight in non presyncopal subjects. In contrast to the Neurolab results, development of orthostatic hypotension and presyncopal events reported in astronauts during standing after space flight have been accompanied by attenuated peripheral vasoconstriction and less elevation in plasma concentrations of norepinephrine. The association between circulating norepinephrine (NE) and peripheral vascular resistance in presyncopal astronauts after space flight led to the conclusion that postflight presyncope can be attributed to a combination of inherently low resistance responses, a strong dependence on volume status, and relative hypoadrenergic function. In the present investigation, we used graded levels of lower body negative pressure (LBNP) to produce linear reductions in stroke volume and performed direct measurements of MSNA to test the hypotheses that (1) elevations in MSNA during central hypovolemia are proportional (i.e., linear) with reductions in stroke volume and; (2) that the slope of the stroke volume MSNA relationship will be reduced in presyncopal subjects. PMID- 15806685 TI - The dangerous secrets patients keep. What you don't tell your doctor can imperil your heart's health. PMID- 15806686 TI - New hope for treating small coronary arteries. PMID- 15806687 TI - New diet drug stirs high hopes. But can the pill, rimonabant, liver up to large expectations? PMID- 15806689 TI - The eye: a window to the heart. How an ophthalmologist's exam may head off serious risks. PMID- 15806688 TI - Fighting women's biggest killer. Cardiac threats you should know, and how to counter them. PMID- 15806691 TI - Healthy eating. All Italian. A recipe for good health. PMID- 15806690 TI - Ask the doctors. I had angioplasty and have one stent. My cardiologist says that I don't need to take clopidogrel (Plavix) plus aspirin. He says one or the other is okay. I'm taking clopidogrel, but notice that advertising for the medication suggest coupling it with aspirin. What's your opinion? PMID- 15806692 TI - Back to basics. Cholesterol. PMID- 15806693 TI - Diabetic ketoacidosis. As dangerous as it sounds. Knowing what diabetic ketoacidosis is and how to prevent it can save you a trip to the hospital. PMID- 15806694 TI - Delicious dressings. Tasty, good-for-you toppings for your favorite veggies. PMID- 15806695 TI - Live fit. What's in it for you? Exercise is important for everyone, but now always for the same reasons. PMID- 15806696 TI - Live fit. Sneaking in fitness. Put the "active" into your family's everyday activities. PMID- 15806697 TI - Live fit. Moving into midlife. Short on time and long on stress, 40- and 50 somethings need exercise more than ever. PMID- 15806698 TI - Live fit. Reenergizing retirement. Sometimes, an empty next is the best place to exercise. PMID- 15806699 TI - Research profile. Mom's antibodies. A risk for type 1? Ake Lermark, PhD. PMID- 15806700 TI - Antipsychotics, insulin resistance, and kids. PMID- 15806701 TI - Hot spots and foot ulcer risk. PMID- 15806702 TI - New Medicare benefits for people with diabetes. The new year brings new Medicare benefits and services for people with diabetes. PMID- 15806703 TI - Microbial life in permafrost. AB - Hydrogenotrophic and acetoclastic methanogenesis was measured at temperatures between 5 and -16.5 degrees C with H14CO3- and 14CH3CO2- as substrates in Siberian permafrost soils. The rate of methane formation was reduced approximately 2-fold over the temperature range from 5 to -1.8 degrees C. For the most active sample "a" temperature dependence of CH4, production at negative temperatures was approximately a 100-fold reduction for a range of -1.8 to -16.5 degrees C for both substrates. According to the Arrhenius equation, the activation energy of methane generation from bicarbonate and acetate for the temperature interval -5 to -16.5 degrees C was reduced by a factor of 3 and 1.5, respectively, in comparison with the temperatures above zero. In the experiments we tested the geological time series, showing the ability of microorganisms to carry out redox reactions after thousands to millions years of existence in permafrost. From the Climate Change point of view, it is important that the recovered organisms are quickly involved anew in present-day ecological processes after instances of permafrost thawing, and may be vital in nutrient recycling and in the production and consumption of greenhouse gases over a large portion of the Earth's surface. From an exobiological point of view, the terrestrial permafrost, inhabited by cold adapted microbes and protecting the cells against unfavorable conditions, can be considered as an extraterrestrial model. The methanogenic bacteria and their metabolic end-products found in the Earth's permafrost provide a range of analogues that could be used in the search for possible ecosystems and potential inhabitants on extraterrestrial cryogenic bodies free of oxygen. PMID- 15806704 TI - The potential of the lichen symbiosis to cope with the extreme conditions of outer space II: germination capacity of lichen ascospores in response to simulated space conditions. AB - Complementary to the already well-studied microorganisms, lichens, symbiotic organisms of the mycobiont (fungi) and the photobiont (algae), were used as "model systems" in which to examine the ecological potential to resist to extreme environments of outer space. Ascospores (sexual propagules of the mycobiont) of the lichens Fulgensia bracteata, Xanthoria elegans and Xanthoria parietina were exposed to selected space-simulating conditions (up to 16 h of space vacuum at 10(-3) Pa and UV radiation at 160 nm < or = lambda < or = 400 nm), while embedded in the lichen fruiting bodies. After exposure, the ascospores were discharged and their viability was tested as germination capacity on different culture media including those containing Mars regolith simulant. It was found that (i) the germination rate on media containing Mars regolith simulant was as high as on other mineral-containing media, (ii) if enclosed in the ascocarps, the ascospores survived the vacuum exposure, the UV-irradiation as well as the combined treatment of vacuum and UV to a high degree. In general, 50 % or more viable spores were recovered, with ascospores of X. elegans showing the highest survival. It is suggested that ascospores inside the ascocarps are well protected by the anatomical structure, the gelatinous layer and the pigments (parietin and carotene) against the space parameters tested. PMID- 15806705 TI - Biological UV dosimeters in simulated space conditions. AB - Polycrystalline uracil thin layers participate in the phage and uracil response (PUR) experiment, assigned to the biological dosimetry of the extraterrestrial solar radiation on the International Space Station (ISS). In ground based experiments (experiment verification tests), the following space parameters were simulated and studied: temperature, vacuum and short wavelength UV (UV-C, down to 200 nm) radiation. The closed uracil samples proved to be vacuum-tight for 7 days. In the tested temperature range (from -20 to +40 degrees C) the uracil samples are stable. The kinetic of dimer formation (dimerization) and reversion (monomerization) of uracil dimers due to short wavelength UV radiation was detected, the monomerization efficiency of the polychromatic deuterium lamp is higher than that of the germicidal lamp. A mathematical model describing the kinetic of monomerization-dimerization was constructed. Under the influence of UV radiation the dimerization-monomerization reactions occur simultaneously, thus the additivity law of the effect of the various wavelengths is not applicable. PMID- 15806706 TI - Simulation experiments of the effect of space environment on bacteriophage and DNA thin films. AB - The main goal of PUR experiment (phage and uracil response) is to examine and quantify the effect of specific space conditions on nucleic acid models. To achieve this an improved method was elaborated for the preparation of DNA and bacteriophage thin films. The homogeneity of the films was controlled by UV spectroscopy and microscopy. To provide experimental evidence for the hypothesis that interplanetary transfer of the genetic material is possible, phage T7 and isolated T7 DNA thin films have been exposed to selected space conditions: intense UVC radiation (lambda=254 nm) and high vacuum (10(-4) Pa). The effects of DNA hydration, conformation and packing on UV radiation damage were examined. Characteristic changes in the absorption spectrum, in the electrophoretic pattern of DNA and the decrease of the amount of PCR products have been detected indicating the photodamage of isolated and intraphage DNA. PMID- 15806707 TI - Calcium-tracers disclose the site of biomineralization in inner ear otoliths of fish. AB - Since changing gravity (concerning direction and amplitude) strongly affects inner ear otolith growth and otolithic calcium incorporation in developing fish, it was the aim of the present study to locate the site of mineralization in order to gain cues and insights into the provenance of the otoliths inorganic compounds. Therefore, larval cichlid fish (Oreochromis mossambicus) were incubated in the calcium-tracer alizarin complexone (AC; red fluorescence). After maintenance in aquarium water for various periods (1, 2, 3, 6, 9 and 12 h; 1, 2, 3, 5, 6, 7, 15, 29, 36 and 87 d), the animals were incubated in the calcium tracer calcein (CAL; green fluorescence). AC thus labeled calcium being incorporated at the beginning of the experiment and would subsequently accompany calcium in the course of a possible dislocation, whereas CAL visualized calcium being deposited right at the end of the test. Subsequently, the otoliths were analyzed using a laser scanning microscope and it was shown that the initial site of calcium incorporation was located directly adjacent to the sensory epithelium and the otolithic membrane. Later, calcium deposits were also found on further regions of the otoliths' surface area, where they had been shifted to in the course of dislocation. This finding strongly indicates that the sensory epithelium plays a prominent role in otolithic biomineralization, which is in full agreement with an own electron microscopical study [ELGRA News 23 (2003) 63]. PMID- 15806708 TI - Efficacy of an ototoxic aminoglycoside (gentamicin) on the differentiation of the inner ear of cichlid fish. AB - Previous investigations revealed that the growth of fish inner ear otoliths depends on the amplitude and the direction of gravity, thus suggesting the existence of a (negative) feedback mechanism. In the course of these experiments, it was shown that altered gravity both affected otolith size (and thus the provision of the proteinacious matrix) as well as the incorporation of calcium. It is hitherto unknown, as of whether sensory hair cells are involved either in the regulation of otolith growth or in the provision of otolithic material (such as protein or inorganic components) or even both. The ototoxic aminoglycoside gentamicin (GM) damages hair cells in many vertebrates (and is therefore used for the treatment of Meniere's disease in humans). The present study was thus designed to determine as of whether vestibular sensory cells are needed for otolith growth by applying GM in order to induce a (functionally relevant) loss of these cells. Developing cichlid fish Oreochromis mossambicus were therefore immersed in 120 mg/l GM for 10 or 21 days. At the beginning and at the end of the experimental periods, the fish were incubated in the calcium-tracer alizarin complexone (AC). After the experiment, otoliths were dissected and the area grown during GM-exposure (i.e., the area enclosed by the two AC labellings) was determined planimetrically. The results showed that incubating the animals in a GM-solution had no effect on otolith growth, but the development of otolith asymmetry was affected. Ultrastructural examinations of the sensory hair cells revealed that they had obviously not been affected by GM-treatment (no degenerative morphological features observed). Overall, the present results suggest that hair cells are not affected by GM concerning their possible role in (general) otolith growth, but that these cells indeed might have transitionally been impaired by GM resulting in a decreased capacity of regulating otolith symmetry. PMID- 15806709 TI - Environmental impacts on the developing CNS: CD15, NCAM-L1, and GFAP expression in rat neonates exposed to hypergravity. AB - We have previously reported that the developing rat cerebellum is affected by hypergravity exposure. The effect is observed during a period of both granule and glial cell proliferation and neuronal migration in the cerebellum and coincides with changes in thyroid hormone levels. The present study begins to address the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of cerebellar proteins that are known to be directly involved in cell-cell interactions [protein expressing 3 fucosyl-N-acetyl-lactosamine antigen (CD15), neuronal cell adhesion molecule (NCAM-L1)] and those that affect cell-cell interactions indirectly [glial fibrillary acidic protein (GFAP)] in rat neonates exposed to centrifuge-produced hypergravity. Cerebellar mass and protein expression in rat neonates exposed to hypergravity (1.5 G) from gestational day (G) 11 to postnatal day (P) 30 were compared at one of six time points between P6 and P30 against rat neonates developing under normal gravity. Proteins were analyzed by quantitative western blots of cerebellar homogenates prepared from male or female neonates. Cerebellar size was most clearly reduced in male neonates on P6 and in female neonates on P9, with a significant gender difference; differences in cerebellar mass remained significant even when change in total body mass was factored in. Densitometric analysis of western blots revealed both quantitative and temporal changes in the expression of selected cerebellar proteins that coincided with changes in cerebellar mass and were gender-specific. In fact, our data indicated certain significant differences even between male and female control animals. A maximal decrease in expression of CD15 was observed in HG females on P9, coinciding with maximal change in their cerebellar mass. A shift in the time-course of NCAM-L1 expression resulted in a significant increase in NCAM-L1 in HG males on P18, an isolated time at which cerebellar mass does not significantly differ between HG and SC neonates. A maximal decrease in expression of GFAP was observed in HG males on P6, coinciding with maximal change in their cerebellar mass. Altered expression of cerebellar proteins is likely to affect a number of developmental processes and contribute to the structural and functional alterations seen in the CNS developing under altered gravity. Our data suggest that both cerebellar development and its response to gravitational manipulations differ in males and females. PMID- 15806710 TI - Future prospects of anticytokine therapy in chronic heart failure. AB - Several lines of evidence suggest that chronic heart failure is a state of chronic inflammation. Indeed, various pro-inflammatory markers, including the cytokines TNF-alpha, and interleukin 6 and 1, are activated in the course of the disease. In chronic heart failure, these substances are frequently induced even before the classical neurohormones angiotensin II and noradrenaline. Although the recently published anti-TNF-alpha trials with etanercept and infliximab have called the beneficial effects of targeting single cytokines into question, the overactive immune system remains a promising target for therapeutic interventions, which aim at slowing down disease progression. Broader approaches are required. These comprise targeting bacterial lipopolysaccharide (endotoxin) that enters the circulation through the oedematous gut wall, immune modulation therapy with patient-derived whole blood exposed to oxidative stress, 3-hydroxy-3 methylglutaryl-coenzyme A reductase inhibitors (the so-called statins) and a number of other substances including pentoxifylline and thalidomide. PMID- 15806711 TI - Outcome measures for clinical trials in neurotrauma. PMID- 15806712 TI - Elevated dietary sodium intake exacerbates myocardial hypertrophy associated with cardiac-specific overproduction of angiotensin II. AB - INTRODUCTION/HYPOTHESIS: Cardiac hypertrophy is an independent risk factor predictive of cardiovascular disease and is significantly associated with morbidity and mortality. The mechanism by which angiotensin II (Ang II) and dietary sodium exert additive effects on the development of cardiac hypertrophy is unclear. The goal of this study was to evaluate the hypothesis that, where there is a genetic predisposition to Ang II-dependent hypertrophy, there is also an increased susceptibility to sodium-induced hypertrophy mediated by AT1 receptor expression. METHODS: Diets of low sodium (LS, 0.3% w:w) and high sodium (HS, 4.0% w:w) content were fed to adult (age 25 weeks) control wild-type mice (WT) and to transgenic mice exhibiting cardiac-specific overexpression of angiotensinogen (TG). At the conclusion of a 40-day treatment period, cardiac tissue weights were compared and the relative expression levels of Ang II receptor subtypes (AT(1A) and AT(2)) were evaluated using RT-PCR. RESULTS: WT and TG mice fed HS and LS diets maintained comparable weight gains during the treatment period. The normalised heart weights of TG mice were elevated compared to WT, and the extent of the increase was greater for mice maintained on the HS diet treatments (WT 12% vs TG 41% increase in cardiac weight index). While a similar pattern of growth was observed for ventricular tissues, the atrial weight parameters demonstrated an additional significant effect of dietary sodium on tissue weight, independent of animal generic type. No differences in the relative (GAPDH normalised) expression levels of AT(1A)- and AT(2)-receptor mRNA were observed between diet or animal generic groups. CONCLUSION: This study demonstrates that, where there is a pre-existing genetic condition of Ang II dependent cardiac hypertrophy, the pro-growth effect of elevated dietary sodium is selectively augmented. In TG and WT mice, this effect was evident with a relatively short dietary treatment intervention (40 days). Evaluation of the levels of Ang II receptor mRNA further demonstrated that this differential growth response was not associated with an altered relative expression of either AT(1A)- or AT(2)-receptor subtypes. The cellular mechanistic bases for this specific ANG II-dietary sodium interaction remain to be elucidated. PMID- 15806713 TI - Effects of concurrent therapy with policosanol and omega-3 fatty acids on lipid profile and platelet aggregation in rabbits. AB - BACKGROUND: Policosanol is a mixture of high-molecular-weight aliphatic primary alcohols isolated from sugarcane wax with cholesterol-lowering and antiplatelet effects. Omega-3 fatty acids (FA) from fish oil can protect against coronary disease. An antiarrhythmic mechanism is emerging as the most convincing explanation for omega-3 FA cardiovascular protection, but triglyceride (TG) lowering effects and inhibition of platelet function could play a role. In view of the effects of policosanol and omega-3 FA on lipid profile and platelet function, potential benefits of combined therapy were expected. OBJECTIVE: To investigate whether combined therapy with policosanol and omega-3 FA would offer some benefit, compared with policosanol or omega-3 FA alone, on serum lipid profile and platelet aggregation in rabbits. METHODS: Male rabbits were randomly distributed in four groups (n = 9 per group). A control group received vehicle, one group was treated with policosanol 5 mg/kg and one with omega-3 FA (eicosapentaenoic acid; EPA [47.0%], docosahexaenoic acid; DHEA [41%]) 250 mg/kg, and the fourth received policosanol 5 mg/kg + omega-3 FA 250 mg/kg. Treatments were orally administered for 60 days. Bodyweight, food consumption and animal behaviour were performed at baseline and study completion. RESULTS: Policosanol significantly lowered low-density lipoprotein cholesterol (LDL-C) [42.7%; p < 0.01] and total cholesterol (TC) [29.4%; p < 0.05], increased high-density lipoprotein (HDL-C) [15.4%; p < 0.05], but left TG levels unchanged. Omega-3 FA significantly lowered TG (47.1%; p < 0.05), but left TC, LDL-C and HDL-C unchanged. Combined therapy decreased LDL-C (38.7%; p < 0.05). Changes in TC, LDL C and HDL-C obtained with combined therapy were greater (p < 0.05) than those with omega-3 FA, but similar to those with policosanol, whereas the opposite applied to TG reduction. No significant changes in lipid profile were observed in the control group. Policosanol and omega-3 FA significantly (p < 0.05) but moderately inhibited platelet aggregation induced with arachidonic acid (13.3% and 12.4%, respectively); combined therapy achieved greater inhibition (23.9%; p < 0.05). All groups showed similar food consumption and bodyweight gain. No toxic signs were observed in any animal. CONCLUSIONS: Concurrent therapy with policosanol 5 m/kg and omega-3 FA 250 mg/kg lowered LDL-C, TC and TG and increased HDL-C. All treatments inhibited platelet aggregation, but better effects were observed with policosanol + omega-3 FA compared with either treatment alone. Combined therapy was well tolerated. These results suggest that treatment with policosanol + omega-3 FA could be useful for regulating lipid profile and inhibiting platelet aggregation, but conclusive demonstration of such effects requires further experimental and clinical studies. PMID- 15806719 TI - President's message. Organized dentistry, dental hygiene must find common ground. PMID- 15806721 TI - Testing your diagnostic skills. Lingual thyroid nodule. PMID- 15806723 TI - Local anaesthetics--Nupercaine and Amethcaine. PMID- 15806728 TI - Estimating quartz dose-response. PMID- 15806729 TI - Oral tolerance: overview and historical perspectives. AB - Oral tolerance was first detailed almost 100 years ago, and since then, it has been shown repeatedly that feeding a wide variety of nonpathogenic antigens can inhibit subsequent systemic immune responses. All systemic immune responses are susceptible, but the degree and scope of the suppression depends on the nature and dose of the fed antigen. Oral tolerance has been described in most mammals, including humans, and it may be the homeostatic mechanism that prevents hypersensitivity to food antigens, as is found in celiac disease. A similar process may prevent the aberrant immune responses to commensal bacteria that occur in inflammatory bowel disease. The ability of oral tolerance to modulate experimental models of autoimmune and inflammatory disease has led to clinical trials in such diseases as rheumatoid arthritis, multiple sclerosis, and type I diabetes, with only variable success. Despite intense research, the exact mechanisms responsible for the systemic tolerance and the reasons why tolerance is the default response to many fed antigens remain controversial. Early studies suggested that CD8(+) "suppressor" T cells were important, but it is now accepted that it may involve either anergy/deletion of CD4(+) T cells, or the induction of regulatory CD4(+) T cells that produce IL-10 and/or TGFbeta. There may also be a role for CD4(+) CD25(+) T(reg), but how and when all these different mechanisms operate is still unclear. The ability of fed antigens to induce tolerance probably reflects their uptake by "quiescent" antigen-presenting cells in the intestine, with presentation to specific CD4(+) T cells in the absence of costimulation, or with the involvement of inhibitory costimulatory molecules. Dendritic cells in the Peyer's patches or mucosal lamina propria are the most likely APCs involved, but it remains to be determined exactly where these interactions occur and what the precise nature of the relevant dendritic cells is. PMID- 15806730 TI - Bargaining or biology? The history and future of paternity law and parental status. PMID- 15806731 TI - The implementation of game in a 20-day head-down tilting bed rest experiment upon mood status and neurotic levels of rest subjects. AB - This study evaluated the effect of the implementation of game on mental health among participants in a bedrest (BR) experiment. Subjects were 12 healthy males aged 20-26, who participated in a 20-day 6-degrees head-down tilting BR experiment. The participants were asked to complete psychometrical questionnaires before, during, and after the experiment. We entrusted the participants to manage their leisure time and they intended a game in which all of them could take part over the experiment period. The general conversation and light-hearted mood among the subjects continued during the experimental period. Longitudinal data analysis showed that levels of neurosis and mood status did not deteriorate during the experiment, while our previous experiments, which were performed under the same protocol as this study except for the implementation of the game showed a distinct deterioration in psychosocial status. We consider that the implementation of game autonomously contributes to the positive effects on the mental health among the participants. PMID- 15806732 TI - History repeating. PMID- 15806733 TI - Reactive oxygen species and antioxidant enzymes activity of Anabaena sp. PCC 7120 (Cyanobacterium) under simulated microgravity. AB - It was found that reactive oxygen species in Anabaena cells increased under simulated microgravity provided by clinostat. Activities of intracellular antioxidant enzymes, such as superoxide dismutase, catalase were higher than those in the controlled samples during the 7 days' experiment. However, the contents of glutathione [correction of gluathione], an intracellular antioxidant, decreased in comparison with the controlled samples. The results suggested that microgravity provided by clinostat might break the oxidative/antioxidative balance. It indicated a protective mechanism in algal cells, that the total antioxidant system activity increased, which might play an important role for algal cells to adapt the environmental stress of microgravity. PMID- 15806734 TI - Short duration microgravity experiments in physical and life sciences during parabolic flights: the first 30 ESA campaigns. AB - Aircraft parabolic flights provide repetitively up to 20 s of reduced gravity during ballistic flight manoeuvres. Parabolic flights are used to conduct short microgravity investigations in Physical and Life Sciences, to test instrumentation and to train astronauts before a space flight. The European Space Agency (ESA) has organized since 1984 thirty parabolic flight campaigns for microgravity research experiments utilizing six different airplanes. More than 360 experiments were successfully conducted during more than 2800 parabolas, representing a cumulated weightlessness time of 15 h 30 m. This paper presents the short duration microgravity research programme of ESA. The experiments conducted during these campaigns are summarized, and the different airplanes used by ESA are shortly presented. The technical capabilities of the Airbus A300 'Zero G' are addressed. Some Physical Science, Technology and Life Science experiments performed during the last ESA campaigns with the Airbus A300 are presented to show the interest of this unique microgravity research tool to complement, support and prepare orbital microgravity investigations. PMID- 15806735 TI - Schedules of controlled substances: placement of Zopiclone into schedule IV. Final rule. AB - With the issuance of this final rule, the Deputy Administrator of the Drug Enforcement Administration (DEA) places the substance, zopiclone, including its salts, isomers and salts of isomers into Schedule IV of the Controlled Substances Act (CSA). As a result of this rule, the regulatory controls and criminal sanctions of Schedule IV will be applicable to the manufacture, distribution, dispensing, importation and exportation of zopiclone and products containing zopiclone. PMID- 15806736 TI - Initial characterization of the microgravity environment of the international space station: increments 2 through 4. AB - The primary objective of the International Space Station (ISS) is to provide a long-term quiescent environment for the conduct of scientific research for a variety of microgravity science disciplines. This paper reports to the microgravity scientific community the results of an initial characterization of the microgravity environment on the International Space Station for increments 2 through 4. During that period almost 70,000 hours of station operations and scientific experiments were conducted. 720 hours of crew research time were logged aboard the orbiting laboratory and over half a terabyte of acceleration data were recorded and much of that was analyzed. The results discussed in this paper cover both the quasi-steady and vibratory acceleration environment of the station during its first year of scientific operation. For the quasi-steady environment, results are presented and discussed for the following: the space station attitudes Torque Equilibrium Attitude and the X-Axis Perpendicular to the Orbital Plane; station docking attitude maneuvers; Space Shuttle joint operation with the station; cabin de-pressurizations and the station water dumps. For the vibratory environment, results are presented for the following: crew exercise, docking events, and the activation/de-activation of both station life support system hardware and experiment hardware. Finally, a grand summary of all the data collected aboard the station during the 1-year period is presented showing where the overall quasi-steady and vibratory acceleration magnitude levels fall over that period of time using a 95th percentile benchmark. PMID- 15806737 TI - The physician's bias. PMID- 15806738 TI - Russian system of countermeasures on board of the International Space Station (ISS): the first results. AB - The system of countermeasures used by Russian cosmonauts in space flights on board of International Space Station (ISS) was based on the developed and tested in flights on board of Russian space stations. It included as primary components: physical methods aimed to maintain the distribution of fluids at levels close to those experienced on Earth; physical exercises and loading suits aimed to load the musculoskeletal and the cardiovascular systems; measures that prevent the loss of fluids, mainly, water-salt additives which aid to maintain orthostatic tolerance and endurance to gravitational overloads during the return to Earth; well-balanced diet and medications directed to correct possible negative reactions of the body to weightlessness. Fulfillment of countermeasure's protocols inflight was thoroughly controlled. Efficacy of countermeasures used were assessed both in- and postflight. The results of studies showed that degrees of alterations recorded in different physiological systems after ISS space flights in Russian cosmonauts were significantly higher than those recorded after flights on the Russian space stations. This phenomenon was caused by the failure of the ISS crews to execute fully the prescribed countermeasures' protocols which was as a rule excused by technical imperfectness of exercise facilities, treadmill TVIS particularly. PMID- 15806739 TI - Use of ozone-depleting substances; removal of essential-use designations. Final rule. AB - The Food and Drug Administration (FDA) is amending its regulation on the use of ozone-depleting substances (ODSs) in self-pressurized containers to remove the essential-use designations for albuterol used in oral pressurized metered-dose inhalers (MDIs). Under the Clean Air Act, FDA, in consultation with the Environmental Protection Agency (EPA), is required to determine whether an FDA regulated product that releases an ODS is an essential use of the ODS. Two albuterol MDIs that do not use an ODS have been marketed for more than 3 years. FDA has determined that the two non-ODS MDIs will be satisfactory alternatives to albuterol MDIs containing ODSs and is removing the essential-use designation for albuterol MDIs as of December 31, 2008. Albuterol MDIs containing an ODS cannot be marketed after this date. PMID- 15806740 TI - A status report on the characterization of the microgravity environment of the International Space Station. AB - A primary objective of the International Space Station is to provide a long-term quiescent environment for the conduct of scientific research for a variety of microgravity science disciplines. Since continuous human presence on the space station began in November 2000 through the end of Increment-6, over 1260 hours of crew time have been allocated to research. However, far more research time has been accumulated by experiments controlled on the ground. By the end of the time period covered by this paper (end of Increment-6), the total experiment hours performed on the station are well over 100,000 hours (Expedition 6 Press Kit: Station Begins Third Year of Human Occupation, Boeing/USA/NASA, October 25, 2002). This paper presents the results of the on-going effort by the Principal Investigator Microgravity Services project, at NASA Glenn Research Center, in Cleveland, Ohio, to characterize the microgravity environment of the International Space Station in order to keep the microgravity scientific community apprised of the reduced gravity environment provided by the station for the performance of space experiments. This paper focuses on the station microgravity environment for Increments 5 and 6. During that period over 580 Gbytes of acceleration data were collected, out of which over 34,790 hours were analyzed. The results presented in this paper are divided into two sections: quasi-steady and vibratory. For the quasi-steady analysis, over 7794 hours of acceleration data were analyzed, while over 27,000 hours were analyzed for the vibratory analysis. The results of the data analysis are presented in this paper in the form of a grand summary for the period under consideration. For the quasi steady acceleration response, results are presented in the form of a 95% confidence interval for the station during "normal microgravity mode operations" for the following three attitudes: local vertical local horizontal, X-axis perpendicular to the orbit plane and the Russian torque equilibrium attitude. The same analysis was performed for the station during "non-microgravity mode operations" to assess the station quasi-steady acceleration environment over a long period of time. The same type of analysis was performed for the vibratory, but a 95th percentile benchmark was used, which shows the overall acceleration magnitude during Increments 5 and 6. The results, for both quasi-steady and vibratory acceleration response, show that the station is not yet meeting the microgravity requirements during the microgravity mode operations. However, it should be stressed that the requirements apply only at assembly complete, whereas the results presented below apply up to the station's configuration at the end of Increment-6. PMID- 15806741 TI - Analysis of a spacecraft life support system for a Mars mission. AB - This report summarizes a trade study conducted as part of the Fall 2002 semester Spacecraft Life Support System Design course (ASEN 5116) in the Aerospace Engineering Sciences Department at the University of Colorado. It presents an analysis of current life support system technologies and a preliminary design of an integrated system for supporting humans during transit to and on the surface of the planet Mars. This effort was based on the NASA Design Reference Mission (DRM) for the human exploration of Mars [NASA Design Reference Mission (DRM) for Mars, Addendum 3.0, from the world wide web: http://exploration.jsc.nasa.gov/marsref/contents.html.]. The integrated design was broken into four subsystems: Water Management, Atmosphere Management, Waste Processing, and Food Supply. The process started with the derivation of top-level requirements from the DRM. Additional system and subsystem level assumptions were added where clarification was needed. Candidate technologies were identified and characterized based on performance factors. Trade studies were then conducted for each subsystem. The resulting technologies were integrated into an overall design solution using mass flow relationships. The system level trade study yielded two different configurations--one for the transit to Mars and another for the surface habitat, which included in situ resource utilization. Equivalent System Mass analyses were used to compare each design against an open-loop (non-regenerable) baseline system. PMID- 15806742 TI - Managed care mandates fall short of curbing California Medicaid costs. PMID- 15806743 TI - The flight experiment ANITA--a high performance air analyser for manned space cabins. AB - Analysing Interferometer for Ambient Air (ANITA) is a flight experiment as precursor for a permanent continuous trace gas monitoring system on the International Space Station (ISS). For over 10 years, under various ESA contracts the flight experiment was defined, designed, breadboarded and set up. For the safety of the crew, ANITA can detect and quantify quasi on-line and simultaneously 32 trace gases with ppm or sub-ppm detection limits. The self standing measurement system is based on Fourier Transform Infrared Spectrometer (FTIR) technology. The system represents a versatile air monitor allowing for the first time the detection and monitoring of trace gas dynamics of a spacecraft atmosphere. It is envisaged to accommodate ANITA in a Destiny (US LAB) Express Rack on the ISS. The transportation to the ISS is planned with the first ATV 'Jules Verne'. The options are either the Space Shuttle or the Automated Transfer Vehicle. PMID- 15806744 TI - Scan time reduction with an adaptive field of view. AB - In magnetic resonance imaging (MRI), there is always a drive toward reducing the acquisition time. In volume imaging, time is often spent in acquiring data where there exists no signal because the imaging volume is larger than the object. In this paper, a method is presented for scan time reduction using an adaptive field of view (FOV). Multislice images are acquired with the FOV in the phase encoding direction of each slice determined by measurements made on the initial localization survey scan. Depending on the region of interest, an optimized FOV is also determined so that scan time is reduced in comparison to a normal scan while improving image resolution. The method is simple to implement and requires no additional hardware. Typical reductions in scan time are on the order 9-14%. PMID- 15806745 TI - Minimally invasive hip resurfacing. AB - Metal-on-metal hip resurfacing is an emerging surgical alternative for younger patients who are expected to outlive a conventional total hip replacement implant. Minimally invasive hip resurfacing uses a smaller surgical incision and new techniques to expose the hip joint. Possible advantages include less damage to soft tissue, muscle and bone; smaller scars; less blood loss; and shorter hospital stays and rehabilitation. Possible disadvantages include damage to soft tissue, femur fracture, neurovascular damage, implant malposition and a longer operating time. The lack of published or unpublished trials or case reports makes it impossible to draw conclusions about the relative effectiveness of minimally invasive procedures for hip resurfacing. Developments in computer navigation systems and fluoroscopic imaging may permit more widespread use of minimally invasive hip resurfacing in the future. PMID- 15806747 TI - Alemtuzumab for B-cell chronic lymphocytic leukemia. AB - Alemtuzumab, a humanized monoclonal antibody, is thought to destroy cancer cells through immune system stimulation or apoptosis induction (programmed cell death). In case series studies using alemtuzumab as salvage therapy, about a third of patients with B-cell chronic lymphocytic leukemia (B-CLL), who were otherwise refractory to chemotherapy, improved. Anti-tumour activity was also observed when the drug was used as first-line therapy or to treat minimal residual disease. Adverse events associated with alemtuzumab included "first-dose" flu-like symptoms, prolonged lymphopenia with a subsequent increased risk of opportunistic infections and viral reactivation (e.g., cytomegalovirus) and transient cytopenias. Data from randomized controlled trials (RCTs), focusing on clinical outcomes such as survival and patients' quality of life, are needed to accurately assess the harm and benefit of alemtuzumab. PMID- 15806746 TI - Recent NASA research accomplishments aboard the ISS. AB - The activation of the US Laboratory Module "Destiny" on the International Space Station (ISS) in February 2001 launched a new era in microgravity research. Destiny provides the environment to conduct long-term microgravity research utilizing human intervention to assess, report, and modify experiments real time. As the only available pressurized space platform, ISS maximizes today's scientific resources and substantially increases the opportunity to obtain much longed-for answers on the effects of microgravity and long-term exposure to space. In addition, it evokes unexpected questions and results while experiments are still being conducted, affording time for changes and further investigation. While building and outfitting the ISS is the main priority during the current ISS assembly phase, seven different space station crews have already spent more than 2000 crew hours on approximately 80 scientific investigations, technology development activities, and educational demonstrations. PMID- 15806748 TI - Pregabalin for peripheral neuropathic pain. AB - Pregabalin is an anticonvulsant drug that is under review for use in Canada. It was recently approved in the US and Europe for the treatment of adults with peripheral neuropathic pain (NeP). In most short-term randomized controlled trials (RCTs) of pregabalin in patients with diabetic peripheral neuropathy (DPN) and or post-herpetic neuralgia (PHN), there were early and significant decreases in mean pain scores. The number of subjects with > 50% reduction in pain score was increased when pregabalin was compared to placebo. The most common adverse effects were dizziness and sleepiness. Withdrawal due to adverse events was also more frequent with pregabalin than with placebo. While pregabalin appears to be an effective treatment for NeP, there is no evidence that it offers advantages over treatments being used in Canada. PMID- 15806749 TI - Lunar precursor missions for human exploration of Mars--III: studies of system reliability and maintenance. AB - Discussions of future human expeditions into the solar system generally focus on whether the next explorers ought to go to the Moon or to Mars. The only mission scenario developed in any detail within NASA is an expedition to Mars with a 500 day stay at the surface. The technological capabilities and the operational experience base required for such a mission do not now exist nor has any self consistent program plan been proposed to acquire them. In particular, the lack of an Abort-to-Earth capability implies that critical mission systems must perform reliably for 3 years or must be maintainable and repairable by the crew. As has been previously argued, a well-planned program of human exploration of the Moon would provide a context within which to develop the appropriate technologies because a lunar expedition incorporates many of the operational elements of a Mars expedition. Initial lunar expeditions can be carried out at scales consistent with the current experience base but can be expanded in any or all operational phases to produce an experience base necessary to successfully and safely conduct human exploration of Mars. PMID- 15806750 TI - Human missions to Mars: new psychological challenges and research issues. AB - Human exploratory missions to Mars represent the most exciting future vision of human space flight. With respect to the distance to travel and mission duration, these missions will provide unique psychological challenges that do not compare to any other endeavor humans ever have attempted. The present paper presents outcomes of two recent projects sponsored by the European Space Agency--Humex and Reglisse--where these challenges and risks have been analyzed in some detail, and where concepts for future research have been developed. This presentation involves three steps. At first, it will be shown that our current psychological knowledge derived from orbital spaceflight and analogue environments is not sufficient to assess the specific risks of mission into outer space. Secondly, new psychological challenges of missions to Mars will be identified with respect to three different areas: (1) individual adaptation and performance, (2) crew interactions, and (3) concept and methods of psychological countermeasures. Finally, different options and issues of preparatory psychological research will be discussed. PMID- 15806751 TI - Professional misconduct. PMID- 15806752 TI - Internal leak. Type 1 endoleak from a right limb dislocation. PMID- 15806754 TI - Basic data on anthropometric measurements and angular measurements of the hip and knee joints for selected age groups 1-74 years of age. PMID- 15806755 TI - Decayed, missing, and filled teeth among persons 1-74 years. PMID- 15806756 TI - [Measurement of isotope shift by means of laser optogalvanic spectroscopy]. AB - The method of measuring isotope shift by means of laser optogalvanic spectroscopy has been built up. The isotope shifts of copper at 578.2nm, europium at 576.2nm and uranium at 575.8nm have been measured. PMID- 15806757 TI - [Investigation of analytical methods of laser photothermal spectroscopy for trace microchemical analysis]. AB - The purpose of this work is to investigate the analytical characteristics of laser photothermal spectroscopy and discuss the feasibility to performance trace microchemical analysis by analytical methods of laser photothermal spectroscopy. PMID- 15806758 TI - [Collisional energy transfer of the Rb (7 2D) state by ground state K atoms]. AB - In the K-Rb vaper mixtures, the two-photon stepwise excitation of rubidium atoms from the ground state to the state Rb 7 2D is obtained by the use of a rubidium lamp and a cw dye laser. The cross sections for the collisional process Rb (7 2D) +K (4S) -->Rb (5S) +K (7S. 5D) have been measured by observation of the resulting fluorescence. The ratio of K (7S. 5D) to Rb7D fluorescence contains a contribution from K 7S<==>5D mixing. The contribution could effectively be subtracted out using a second experiment in which a potassium lamp and a cw dye laser were used to pump the K7S or 5D state, and the ratio of 7S to 5D or 5D to 7S fluorescence was monitored. The cross sections (in 10(-15)cm2) for Rb7D-->K7S. 5D excitation transfer are 25.8 +/- 10. 3 and 1.5 +/- 0.6 respectively. PMID- 15806759 TI - [Deducing j-j coupling spectroscopic terms by combination-matrix method]. AB - Deducing j-j coupling spectroscopic terms by combination-matrix method was applied for the first time. PMID- 15806760 TI - [The microspectra of chromosome]. AB - The spectra of radicle chromosome of broad bean, marrow cell chromosome and marrow cell micro-nucleus of small white mouse were measured within the range of visible light by microspectrum technique. The differences among the spectra were found. PMID- 15806761 TI - [The developement of the near-field scan optical microscope and near-field spectroscopy]. AB - This paper introduces the basic principles and techniques of the near-field microscope and the status of recent development in the near-field spectroscopy. We also discuss problems facing the analysis of the results of the near-field spectra. PMID- 15806762 TI - [X-ray photoelectron spectroscopy and mass spectrum study of tin and germanium compounds]. AB - The relationship between the chemical displacement of binding energy and the different chemical environment for tin and germanium compounds was studied by means of X-ray photoelectron spectroscopy and mass spectrum. PMID- 15806763 TI - [Absorption spectra study for fullerene-doped in polystyrene film]. AB - The absorption spectra of C60 and C70-doped in polystyrene film and in solution were studied. The experiment results showed that the positions of the absorption peaks for C60-doped in polystyrene film moved a little toward longer wavelength than in n-hexane, and regular solid material appeared while the ratio of C60 doped in the film increased; The absorption peaks for C70-doped in polystyrene film were almost in consistent with that in solution. We suggest that C60 molecules may be connected as "microcrystalline" in polystyrene film, while C70 may be exist as single molecule or particle less than C60 in it. The preliminary model for the growth of "microcrystslline" in polystyrene film has been proposed. PMID- 15806764 TI - [SERS and electrochemistry studies on the structure of thiourea adsorption layer on silver electrode]. AB - The structure of thiourea adsorption layer on silver electrode was studied by SERS and electrochemistry. It was found that the thiourea can be adsorbed on the electrode surface and can inhibit the reaction of hydrogen separation. And a change in structure of the thiourea adsorption layer on electrode surface took place at the voltage of about -0.6V. The further SERS experiment showed that the thiourea adsorption layer has a loose structure at about -0.9V and a compact structure at about -0.3V - -0.5V. The transition region is at about -0.6V - 0.8V. PMID- 15806765 TI - [Preparation and property investigations of nanosized titanium oxide microcrystals]. AB - A naosized TiO2 powder was successfully prepared by the hydrolysis of titanium isopropoxide through sol-gel process. The microstructure of TiO2 powder was investigated by transmission electron microscopy and X-ray diffraction. Both anatase and rutile phases were found in TiO2 powder annealed for 1 hour at 600 degrees C. The characteristics of X-ray photoelectron spectra and infrared spectra could be affected by the size and aggregation state of the microsrystalc constituting the TiO2 powder. PMID- 15806766 TI - [Study on the tribochemistry of antiwear additive BA1 by FTIRM]. AB - This paper studied the tribochemistry of antiwear additive BA1 by FTIR microscope technique. We observed the changes of physical properties, interpreted the infrared spectra of the products in the track and concluded the chemical mechanism through the change of BA1 concentration, load and rubbing time. The results shows that the BA1 was decomposed, oxided and a thin film was produced when rubbed. This film makes the rubbing efficiency less. PMID- 15806767 TI - [Syntheses and spectra of Mn(III)-chlorophyll-a and Mn(II)-chlorophyll-a]. AB - Manganese(III)-chlorophyll-a [Mn(III)-Chl-a] was synthesized by the reaction of pheophytin-a and manganese(II) acetate and separated by HPCL. Manganese(II) chlorophyll-a [Mn(II)-Chl-a] was acquired from Mn(III)-chl-a reduced by sodium hydrosulfite (Na2S2O4) in acetone-water. Because Mn(II)-Chl-a is oxidized very easily in the air, Mn (II)-Chl-a was kept under a nitrogen atmosphere and reductant during the spectroscopic measurements. Elemental analysis, absorption and FT-IR spectra proved that the metal Mn-substituted Mg of chlorophyll-a has been successfully performed and possible structure of new complexes was also given. Their formulae are [Mn(II) (chlorophyll-a)] and [Mn(III) (chlorphyll-a) (CH3COO)-(H2O)] respectively. PMID- 15806768 TI - [Ann applied to the prediction of vibration frequencies of octahedral modes (v1 and v2) of MX6(n-) ions (X = F, Cl, Br, I)]. AB - Artificial neural networks back-propagation algorithm was applied to the prediction of vibration frequencies of v1 and v2 modes of octahedral hexahalide (MX6(n-)). Three-layer networks with one and two output nodes were used. Two inertia terms and training step controlling scheme were adopted to weights adjustment. The result of one output node networks has little difference from that of two output nodes networks. The frequencies of [Mof6]2-, [BiF6]- and [AuF6]- from literature are a great deal different from those calculated or predicted values. PMID- 15806769 TI - [Studies on the spectral characteristics of argon in a novel microwave plasma enhanced glow discharge source]. AB - This paper describes a investigation about the spectral characteristics of argon in a novel microwave plasma enhanced glow discharge source. In different discharge conditions, such as glow discharge, microwave plasma and microwave plasma enhanced glow discharge, there are different distributions of argon spectral lines. PMID- 15806770 TI - [ICP-AES method for the determination of migrated hazard elements in toy materials and coatings]. AB - An ICP-AES method is established for the determination of Sb, As, Ba, Cd, Cr, Pb, Hg and Se in toy materials and coatings under conditions that simulate the situation where the materials stay 4 hours in alimentary tract after swallowing. PMID- 15806771 TI - [Some considerations in the designed scheme of WLY100 sequential scanning ICP spectrometer]. AB - The designed scheme of China made sequential scanning ICP spectrometer was described in this paper. The considerations of monochromater, RE generator, scanning machine, intensity measurement, the utility of far ultra vioiet region, the controlling of temperature and carrier ags flow, and the software were discussed detailly. A good designed seheme promoted the manufacturing and performed in short period. PMID- 15806772 TI - [Determination of atomization efficiency in graphite furnace with V-boat]. AB - Based on that the absorbance is proportional to the effective length of the absorbing layer of atoms, a simple foumula is derived by which the atomization efficiency. Two atomization efficiencies were calculated: beta1 and beta2, which gave the fraction of calculated the total number of analytical atoms that enter the analysis volume during each stage of atomization. It is based on two different calculated ways for the mean residence time of atoms, and the beta1 and beta2 values of Ga and Tl in furnace with V-boat in various atomization temperatures were determined. Influence of the loss process of atomic vapor on the atomization efficiency value was discussed. PMID- 15806773 TI - [Determination of copper, zinc, calcium, magnesium and iron microelements in whole blood samples from human's ear using microsample-injection FAAS]. AB - This paper established a new and convenient method for the determination of Cu, Zn, Ca, Mg and Fe microelements content in whole blood samples from human's ear by using flame AAS with microsample injection. The results show that recoveries of these microelements are above 96.0% and relative standard deviation of Zn, Ca, Mg and Fe are less than 2.0%, Cu is less than 3.9%. Satisfactory results are obtained with only 10 microL whole blood samples from human's ear. PMID- 15806774 TI - [Flame atomic absorption spectrometry determination of zinc and ferrum in enamel]. AB - The amount of zinc and-ferrum in enanmel was determined by flame AAS. For twenty six healthy men, the average of zinc and ferrum were 325.6 +/- 84.8 and 34.7 +/- 9.1 microg/g, respectively. For fifty four patients with dental caries disease, the average of zinc and ferrum were 269.9 +/- 63.5 and 38.7 +/- 10.0 microg/g, respectively. The difference between healthy men and patients of zinc and ferrum rate was insignificant. As to the determination method, the rate of recovery varied from 97.0 to 101.5% for zinc and from 97.2 to 102.4% for ferrum. The RSDs fo Zn and Fe are less than 2.60 and 2.10% respectively. PMID- 15806775 TI - [Studies on determinaion of selenium in drink by GFAAS]. AB - Pd(NO3)2-Triton X-100 was used as matrix modifier for determination of selenium in drink. In this way ashing temperature can be increased up to 1200 degrees C and absorbance can increase by 2.92 times. Interferences of Fe, P and other elements were reduced effectively by means of Zeemen effect background correction. Detection limit is 8.0ng/mL and linear range is from 10 to 120ng/ml, and r = 0.9931. PMID- 15806776 TI - [Three-dimensional microanalysis of arsenic impurity in silicon]. AB - The three-dimensional distribution of arsenic impurity concentration on the silicon wafer surface is determinaed by means of synchrotron radiation X-ray microfluorescence (SRXRF) and total reflection X-ray fluorescence (TXRF) anslysis. The result of depth profile of As is checked with secondary ion mass spectrometry (SIMS) and the agreement is reasonably good. PMID- 15806777 TI - [Fluorescence spectroscopic analysis of huminite of the peat and soft brown coal]. AB - The huminite of the peat and soft brown coal in the Zhenan basin, western Yunnan, has been studied by means of fluorescence microscopy. The study reveals that the fluorescent features of the huminite are considerabably complicated: some huminite, like liptinites, can give out dark yellow to yellow fluorescent light, I546 ranging from 8.13 to 13.17; and the fluorescent spectrum having one peak or two peaks, lambdamax ranging from 555 to 585nm and Q650/500 from 0.60 to 2.93. Most of huminites do not give out fluorescent light or give out dark brown fluorescent light, I546 ranging from 0 to 5.38. Because of having extraordinarily weak fluorescent light, it is very difficult to obtain their fluorescent spectrum. Nevertherless, after being stimulated for several minutes, some huminites that do not give out fluorescent light or give out dark brown fluorescent light may give out yellowish brown to dark orange fluorescent light, I546 ranging from 6.12 to 12.17, lambdamax. from 590 to 615nm and Q650/600 from 3.49 to 4.31. With the stimulating time becoming longer (10 to 20min), some huminites,may give out sharper yellow fluorescent light, I546 ranging from 12.18 to 20.01, lambdamax from 555 to 595nm and Q650/500 from 0.87 to 1.60. PMID- 15806778 TI - [Catalytic fluorimetric determination of trace amount of bromine ion]. AB - A new catalytic fluorimetric method for the determination of bromine ion based on its catalytic effect on the oxidation of fluorescein by potassium bromate in the solution of dilute sulfuric acid has been established. The detection limit for Br is 1.94 x 10(-2) microg/25 mL. The linear range of the determination is 6.0-12.0 microg/25 mL. The method has been applied to the determination of Br- in tablet pyridostigmini bromidum sample with satisfactory results. PMID- 15806779 TI - [Determination of trace selenium in herbs of invigorating circulation blood and removing blood stasis by fluorometry]. AB - Selenium in herbs of invigorating circulation blood and removing blood stasis was analysed by fluorometry. The RSD was less than 4.9%, the rate of recovery varied from 96% to 102%. The method has better selectivity. It is operated simply, and is suited for selenium of herbs analysis. PMID- 15806780 TI - [Determination of prostaglandinum E1 by fluorescence extinction method]. AB - In this paper, we have studied the fluorescence extinction effect of methyl alcohol by prostaglandinum E1 (PG E1) and found the linear relation between the fluorescence extinction intensity and prostaglandinum E1 concentration. Its linear equation is deltaF = 0.222c + 9.33 x 10(-2)(deltaF: fluorescence extinction value; c: PG E1 concentration microg/mL), correlation coeficient r = 0.99930. The results derived from fluorescence extinction method correspond to the results from violet spectrophotometric method. PMID- 15806781 TI - [Determinaion of artificial synthetic pigment in the drug by reflection spectrophotometry]. AB - Solid phase spectrophotometry is a new, simple, fast and sensitive trace analytical method. In this report, it is presented a method that the polyamide was used for the absorbence and the reflectances absorbances were measured directly with remodelled the colorimetric cell. Not only the trace inorganic ion can be analysed by the solid phase method, but also the artificial synthetic pigment in the drug can be determined, the results are satisfied. PMID- 15806782 TI - [Study of conditions on determination of micro phosphorous by synchronous fluorescence quenching of rhodamine B]. AB - The fluorescence behavior of the system and the conditions of the title method has been studied. Non-ionic surfactant op was added to increase the sensitive and stability of the determination. This method has been applied to the determinations of phosphorous in some black foods. Both the accuracy and precision are satisfactory. PMID- 15806783 TI - [Dual wavelength multiplication subtractive spectrophotometric determination of sulfadiazinum and trimethoprimum in compound sulfadiazine tablets]. AB - This paper described the simultaneous determination of sulfadiazine (SD) and trimethoprim (TMP) in tablets co-Trimediazini by dual wavelength spectrophotometry multriplication subtractive method. This method was based on the measurement of both absorbance of standard solution of SD (8-14) x 10( 6)g/mL, TMP (1-2) x 10(-6)g/mL and sample solution at wavelength 242.5 nm and 228.0nm. The average recovery and RSD of SD and TMP were 100.60%, 0.33% and 98.27%, 1.58%, respectively (n=20). The method was simple, quick and results satisfactory. PMID- 15806784 TI - IUNS in the twenty first century on the shoulders of the twentieth century giants of nutrition. PMID- 15806785 TI - Nutrition in the third millennium: countries in transition. PMID- 15806786 TI - A vision of the nutritional sciences in the third millennium. PMID- 15806787 TI - Vitamin A deficiency and the global response. PMID- 15806788 TI - Food carotenoids: analysis, composition and alterations during storage and processing of foods. AB - Substantial progress has been achieved in recent years in refining the analytical methods and evaluating the accuracy of carotenoid data. Although carotenoid analysis is inherently difficult and continues to be error prone, more complete and reliable data are now available. Rather than expressing the analytical results as retinol equivalents, there is a tendency to present the concentrations of individual carotenoids, particularly beta-carotene, beta-cryptoxanthin, alpha carotene, lycopene, lutein and zeaxanthin, carotenoids found in the human plasma and considered to be important to human health in terms of the provitamin A activity and/or reduction of the risk for developing degenerative diseases. With the considerable effort directed to carotenoid analysis, many food sources have now been analyzed in different countries. The carotenoid composition of foods vary qualitatively and quantitatively. Even in a given food, compositional variability occurs because of factors such as stage of maturity, variety or cultivar, climate or season, part of the plant consumed, production practices, post-harvest handling, processing and storage of food. During processing, isomerization of trans-carotenoids, the usual configuration in nature, to the cis forms occurs, with consequent alteration of the carotenoids' bioavailability and biological activity. Isomerization is promoted by light, heat and acids. The principal cause of carotenoid loss during processing and storage of food is enzymatic or non-enzymatic oxidation of the highly unsaturated carotenoid molecules. The occurrence and extent of oxidation depends on the presence of oxygen, metals, enzymes, unsaturated lipids, prooxidants, antioxidants; exposure to light; type and physical state of the carotenoids present; severity and duration of processing; packaging material; storage conditions. Thus, retention of carotenoids has been the major concern in the preparation, processing and storage of foods. However, in recent years the effect of processing on bioavailability has been focalized. More than a century after their discovery, carotenoids continue to be intensely investigated in various areas. This article aims to give an overview of current knowledge in Food Science and Technology, which has bearing on the role of carotenoids in human health. PMID- 15806789 TI - The significance of vitamin A for the development and function of the lung. PMID- 15806790 TI - Vitamin E, on the biopotency of gamma-tocopherol. PMID- 15806791 TI - Vitamin C: from popular food supplement to specific drug. AB - The daily requirement of a human person for vitamin C (ascorbic acid) has now been established at 100 mg. This value was already on the map when Arnold Durig put together the most important needs of nutritional ingredients. The modern value rests on the saturating level of ascorbate in leukocytes, which is in the millimolar range. The mechanism of accumulation of ascorbate in these cells rests on the uptake of oxidized dehydroascorbic acid. It is very efficient and avoids loss of vitamin which occurs in vitro when ascorbate is oxidized because of the great instability of the dehydro form. Therefore and increased requirement in case of infection is very unlikely from the biochemical point of view. However, low concentrations of ascorbate are found in patients suffering from arterial diseases or diseases accompanied by arterial damage such as diabetes mellitus. Ascorbate is known as a protection factor for the arterial endothel, but it is not clear by what mechanism this protection is brought about. Moreover, under clinical conditions very high concentrations are needed, which are achieved only by intravenous infusion, and the protection is only observed when the disease is manifest, not in healthy people. Therefore, also in this respect an increase in daily intake seems of no prophylactic value. Thus, by using high concentrations of ascorbate as an i.v. drug, effects of this substance frequently observed in vitro, could be used for therapy. This includes not only treatment of arterial diseases, but also relates to the cytotoxic effects of the vitamin against certain tumor cells and may assist conventional chemotherapy. PMID- 15806792 TI - Folate, vitamin B12 and birth defects. AB - While the evidence that folic acid/folate can prevent most cases of NTDs if taken periconceptually, the position is less clear with respect to OFCs. Some studies indicate a positive protection while others are negative. There are also conflicting reports as to whether or not the C677T variant of MTHFR increases risk. This is to be contrasted with NTDs where it clearly does. PMID- 15806793 TI - What have animal studies told us about the role of folic acid and vitamin B12 in health? PMID- 15806794 TI - Toxicological aspects of iron. PMID- 15806795 TI - Anaemia--a matter of serious concern and threat to human development. PMID- 15806796 TI - Calcium, iron, zinc, copper, manganese and selenium concentrations in typical hospital diets. PMID- 15806798 TI - Thyroid fetomaternal relationship in iodine deficiency. PMID- 15806797 TI - Zinc deficiency: issues for public health, assessment, and prevention. PMID- 15806799 TI - Dietary fatty acids in an era of genetic modification. PMID- 15806800 TI - Dietary fat in the prevention of coronary heart disease. PMID- 15806801 TI - Essential fatty acids in health and chronic diseases. PMID- 15806802 TI - Long-chain polyunsaturated fatty acids in visual and neural development: cellular and molecular mechanisms. PMID- 15806803 TI - Considerations on sources of milk fatty acids: investigations of kinetics and concentrations. PMID- 15806804 TI - Methods for determining the metabolism of alpha-linolenic acid in human subjects from intrinsically-labeled food. PMID- 15806805 TI - Contents and analytical aspects of trans and conjugated fatty acids especially in food. PMID- 15806806 TI - Effect of cis-9,trans-11 and trans-10,cis-12-CLA isomers in two strains of mice. PMID- 15806807 TI - Nutritional aspects of glutamate. PMID- 15806808 TI - Glutamate metabolism in the gut. PMID- 15806809 TI - The role of glutamate in fetal hepatic glucogenesis. PMID- 15806810 TI - Changes in selection and intake of foods in humans, induced by the addition of monosodium glutamate. PMID- 15806811 TI - Brain mechanisms that analyse umami taste and their relation to the control of feeding. PMID- 15806812 TI - Monosodium glutamate (MSG) and taste-mGluR4, a candidate for an umami taste receptor. PMID- 15806813 TI - Microbial synthesis of essential amino acids in the human colon--an unsolved problem. PMID- 15806814 TI - On the recommended nutrient and energy intakes for the European Community. PMID- 15806815 TI - New reference values for nutrient intake in Germany, Austria and Switzerland (DACH-Reference Values). PMID- 15806816 TI - Genomic stability: a new paradigm for recommended dietary allowances (RDAs). PMID- 15806817 TI - Healthy lifestyles and health promotion: a challenge and an opportunity for nutrition science. PMID- 15806818 TI - The EURODIET initiative and health promotion prospects: the case of Greece. PMID- 15806819 TI - National food consumption survey in children aged 1-9 years: South Africa 1999. PMID- 15806820 TI - Vital statistical implication of nutrition in Hungary. PMID- 15806821 TI - Towards institutionalisation of community-based actions to improve child nutrition. A review of African experience. PMID- 15806822 TI - Policy and implementation of community-based nutrition program in Thailand. PMID- 15806823 TI - Methodological approach to a multidimensional evaluation of food and nutrition policies. AB - Recommendations arising from global conferences and summits, expressed the need to formulate and implement public policies to improve household food security. In the context of maximizing benefits given available resources, comprehensive evaluations of the nutrition policies and programs are needed. One obstacle to overcome was a clear definition of terminology; the words efficiency, effectiveness, efficacy and impact, widely used in the context of program evaluation, are sometimes interchanged and there is no consensus about their precise definition. Another approach to health evaluation is based in the paradigm structure-process-outcome. The level structure contemplated the installations, resources, instruments (physical and discursive), as well as the technical bureaucratic organizational structures. Process encompassed the whole set of intervention activities developed, whereas the dimension of outcome comprised the effects of interventions on the health and nutrition of beneficiaries. Each of the three dimensions constitutes a continuum in the evaluation procedure: the structure only fulfills its purposes if the processes are adequate and conversely, processes cannot alone supersede structural limitations. Moreover, all the three dimensions only reach their ultimate objectives through the completion of outcomes. The methodology proposed here has been tested in the case of Bahia, Northeast Brazil, and it was found to be adequate for this type of analysis. We hope this approach of a comprehensive multidimensional evaluation, constitutes an effective contribution for program planners and program managers, in particular with regard to the obstacles detected, some of which can surely be overcome. PMID- 15806824 TI - From needs-based to rights-based approaches to child nutrition: lessons learnt from the 1990s. PMID- 15806825 TI - Nutrition and poverty reduction: lessons on policy change from the World Bank/UNICEF nutrition assessment. PMID- 15806826 TI - Capacity development initiative in the area of food and nutrition Southern Africa. PMID- 15806827 TI - Better understanding of eating pattern for better understanding of human nutrition. PMID- 15806828 TI - Highlighting the positive impact of increasing feeding frequency on metabolism and weight management. AB - Research on feeding frequency started more than 20 years ago and some studies have shown evidence of nutritional benefits, especially on metabolism and body weight management. Advice on feeding frequency could play an important role in public health policies by reducing levels of overweight and obesity, the prevalence of which has dangerously increased in most countries over the last few decades. The 17th International Congress of Nutrition brought to the forefront the benefits of increasing feeding frequency (i.e. keeping the same total daily energy intake but dividing it into more frequent meals than usual). Recent epidemiological studies, mostly carried out in France, have provided evidence on the beneficial effects of a fourth meal for those individuals who habitually choose this pattern. Supported by metabolic data, these findings have now been supported by experimental studies. The "gouter", commonly eaten in the afternoon in France by most children and many adults, has the biological characteristics of a meal because it is eaten in response to hunger. Suppressing the "gouter" in "habitual fourth meal eaters" soon leads to an increase in Body Mass Index (BMI). Further, people who are regular "gouter" eaters have a higher carbohydrate intake and better metabolic profile than other adults, even though their total energy intake is not greater. Increased feeding frequency leads to a reduction in the total secretion of insulin, an improvement in insulin resistance and a better blood glucose control, as well as an improvement in the blood lipid profile. The experts agreed that, as long as we do not consume more energy than we use up and we only eat when we are hungry, it may be useful to split our total energy intake into as many meals as our social pattern allows. However, the pattern of eating cannot be completely dissociated from the composition of foods consumed. Therefore within this energy intake, we must take care to consume not only a good balance of macronutrients with high carbohydrate and low fat levels, but also ensure that we get an adequate intake of essential micronutrients. "What you eat" and "When you eat it" are public health messages to communicate: frequent consumption of low energy dense high carbohydrate foods, rich in micronutrients, must be encouraged ensuring that energy intakes are not greater than energy expenditures and that eating episodes occur in a hunger state. PMID- 15806829 TI - A perspective on nutrition communication in developing countries: from theory to practice. AB - Though nutrition communities are still far from understanding how to bring science to bear in formulating effective policies and programs, the contributions of nutrition science have provided recommendations leading to ways and means to improve nutrition in developing countries. To transfer these recommendations into successful implementation, effective nutrition communication has played an important role in the development. Nevertheless, its success has been limited due to several factors including the fact that there is likely only a small proportion of professionals who consistently aim to use theory as a tool to guide action. This paper highlights the importance of using theoretical frameworks to guide the design and implementation. Based on Thailand's experience, social cognitive theory and social marketing are suggested for further applications in developing country context. It is proposed that the role of researchers/practitioners should be formed to facilitate nutrition implementation. Issues relating to the creation of supportive structures and environments i.e. quality personnel, capacity building, proper status and reasonable budgets, are also identified. PMID- 15806830 TI - Nutrition communication from theory to practice: some future perspectives. AB - 'Nutrition communication' can be defined as the process by which nutrition knowledge is converted into dietary change. Nutrition communication then includes nutrition education--the process by which people are informed, and sometimes empowered by, nutrition information--but also other actions taken to improve peoples' diets such as restricting misinformation about nutrition or manipulating the composition, availability or price of foods. Dietary goals for populations and food-based dietary guidelines for individuals constitute the starting point for nutrition communication, and if these could be more evidence-based we would be in a much stronger position to develop more effective nutrition communication. A population-based approach to nutrition communication is complementary to an individualistic approach in theory, but in practice they compete for attention in food policy making circles. PMID- 15806831 TI - Communicating emerging science to maximize understanding. PMID- 15806832 TI - Training in public health nutrition: a conceptual framework. PMID- 15806833 TI - Public health nutrition in Latin America. AB - An inquiry into options for Masters-level training and into attitudes and perceptions among a convenience sample of nutrition students and professionals from 11 countries suggests that the term, "Public Health Nutrition", as such, is poorly represented and poorly understood in the Latin American region. At least six countries (Brazil, Chile, Cuba, Mexico, Peru, Puerto Rico) at seven sites have Masters programs to provide training for nutrition in a public health context or public health with an emphasis in nutrition. Exploring alliances from the Americas with the formal PHN discipline emerging in Europe should enrich the mutual perspective on curriculum design. However, the form and context of postgraduate training in Latin America must consider first and foremost its own job-markets, diverse public health needs, and resource allocations in building or transforming training programs. PMID- 15806834 TI - The development of a European master programme in public health nutrition. PMID- 15806835 TI - The importance of high-level training for nutrition scientists in Sub-Saharan Africa. AB - Can the African people investigate and critically analyse nutrition needs in their context, identify national or community nutrition problems, plan and evaluate nutrition programs, educate, and communicate scientific knowledge in nutrition without high-level trained staffs with a strong scientific base in human nutrition? Will they be able to tackle their nutrition problems without a sustainable organisation for long-term training and research? In this paper, an attempt is made to analyse factors that will contribute to improving human nutrition in Sub-Saharan Africa. The paper also highlights an experience of institutional capacity building in Senegal, West Africa. PMID- 15806836 TI - A human rights-based approach to food and nutrition in development--reflections from the ACC/SUB-Committee on Nutrition. PMID- 15806837 TI - Mobilising states and other actors for a rights-based approach to food and nutritional health. PMID- 15806838 TI - Nutrient regulation of immune functions. PMID- 15806839 TI - The interaction of nutrition and the immune response: new approaches. PMID- 15806840 TI - Nutritive amino acids--effective modulators of the immune response. PMID- 15806841 TI - Adverse reactions to foods. PMID- 15806842 TI - Peanut allergy: lymphocyte response to peanut extract. PMID- 15806843 TI - Chemical and technological aspects of food allergies. PMID- 15806844 TI - Update on HIV/HAART associated morphologic and metabolic abnormalities. PMID- 15806845 TI - Breastfeeding and risk of HIV transmission: an update. PMID- 15806846 TI - The molecular basis of body weight control. PMID- 15806847 TI - Effects of protein vs. carbohydrate-rich diets on fuel utilisation in obese women during weight loss. AB - BACKGROUND: Energy restriction is a common therapeutic approach for weight reduction, but the most effective macronutrient distribution of the hypoenergetic diet as well as the role of the metabolic processes involved require further investigations. OBJECTIVE: To study the effect of a high protein (HP) hypoenergetic diet compared with a high carbohydrate (HC) hypoenergetic diet on fuel utilisation changes during the weight loss process in obese women. DESIGN: Eleven obese (BMI>30) women were randomly assigned to a 10w dietary intervention study comparing HP (30% protein) or HC (55% carbohydrate) energy restricted diets providing 30% energy fat content. Substrate utilisation was evaluated by indirect calorimetry. Body weight and composition (Bod Pod) and blood measurements were performed before and after weight loss. RESULTS: On average, the individuals on the HP dietary group lost 4.4 kg more than those in the HC program (p<0.50), which was mainly due to a fat mass loss (3.7 kg, p<0.05) with no statistical differences in lean body mass reduction. These losses were accompanied by a significant decrease in fasting leptin in the HP group (-52%; P<0.05). On the other hand post-absorptive lipid oxidation decreased in the HC group (-48%) and remained unchanged in the HP groups. CONCLUSIONS: The replacement of some dietary carbohydrate by protein in energy restricted diets, improves weight and fat losses and specifically promotes lipid oxidation in the fasting state, without major different in lean body mass depletion. PMID- 15806848 TI - Role of nutrition in the management of diabetes mellitus. PMID- 15806849 TI - Dietary fibre and type 2 diabetes mellitus. PMID- 15806850 TI - Nutritional rickets in developing countries. PMID- 15806851 TI - Nutrition and osteoporosis: facts and uncertainties about calcium and vitamin D recommendations. AB - The current calcium recommendations in Europe are lower than in USA. During the growth, although intervention studies had shown bone mass gains with calcium supplementation, these effects did not persist after withdrawal of the supplements and so the long-term effects of calcium supplementation on peak bone mass are not known. Besides, the relative importance of the several factors that influence peak bone mass needs to be established. PMID- 15806852 TI - Treating severe malnutrition effectively: the challenge to all nutritionists. PMID- 15806853 TI - An extended field test of the WHO severe malnutrition guidelines in hospitals in developing countries: preliminary observations. AB - The WHO has published guidelines for the inpatient management of severe malnutrition. A qualitative study in hospitals in developing countries is being conducted to document the re-organisation of the clinical ward and support services required to implement these guidelines and to gain an impression of the feasibility and sustainability of such a re-organisation. Following a postal survey of experts in the management of malnutrition in children in developing countries, hospitals were contacted and asked if they were interested to participate in the study. If so, they were requested to submit background information about admission patterns, the frequency of malnutrition, and current practice. Based on this information, hospitals are selected for a preliminary visit. Following this, and the final selection, a paediatrician conducts three visits to the study hospital over a one-year period to appraise the current practice, assist the health staff in recognising the strengths and shortcomings of their current management, help them find locally appropriate solutions, support the implementation process through a participatory approach and assess the outcome. A structured survey instrument is used to guide the assessment and identification of problems. Results of the first visit, which documents the existing situation, and changes identified by staff and implemented during the second visit are presented. PMID- 15806854 TI - The nature of scientific evidence and its relevance to policy. PMID- 15806855 TI - Translating science into improved health. PMID- 15806856 TI - Nutrition and cancer--translating science into practice. Symposium summary. PMID- 15806858 TI - A collaboration to increase nutrition work in general medical practice and make it more effective. PMID- 15806857 TI - Dietary calcium and colon cancer prevention. PMID- 15806859 TI - Possible lessons for physician counseling on obesity from the progress in smoking cessation in primary care. PMID- 15806860 TI - The current status of parenteral and enteral nutrition in the critically ill. PMID- 15806861 TI - Fat--the good, the bad and the trans. PMID- 15806862 TI - Thiol-mediated redox regulation. PMID- 15806863 TI - Nutritargeting. PMID- 15806864 TI - Nutrition surveys: springboards for action, not academic exercises. PMID- 15806865 TI - The CDC-UCLA USAID national nutrition baseline surveys of the 1970's. PMID- 15806866 TI - Using national health and nutrition surveys for policy and programs: experiences from the demographic and health surveys. AB - National health and nutrition surveys have proved invaluable for policy and programs internationally, nationally and locally. With internationally comparable data, decisions can be made on trends and to monitor progress towards improving the nutritional status of women and children. In addition, the iterative process of survey and indicator development ensures that the use of the findings are fed back into the survey for improved design, data collection and measurements. Researchers, policy makers, program managers and surveyors work together for maternal child health. PMID- 15806867 TI - The challenges of regional program coordination and scientific leadership. PMID- 15806868 TI - Integrating strategies for combating vitamin A deficiency: successes in Asia. AB - Indonesia and Bangladesh were among the first countries to distribute vitamin A capsules (VAC) to underfives in the early 1970s. Since then, VAC coverage has increased very much. In Indonesia, VAC are now distributed to underfives in two special months. In Cambodia, VAC distribution started less than a decade ago and coverage is yet <50%. After being linked to National Immunization Days, it is now linked to Immunization Outreach, which seems a good strategy. Because VAC are only distributed to underfives and postpartum women, and because coverage is usually around 50-80%, other strategies are implemented simultaneously. Recent HKI-evaluations found that promotion of dark-green leafy vegetables and eggs in Indonesia increased their consumption and improved vitamin A status. In Bangladesh, underfives and women of households with a homestead garden and/or poultry had a lower risk of vitamin A deficiency. The HKI Homestead Food Production Program aims at improving homestead food production activities in order to increase production and consumption of vitamin A-rich foods. Food fortification in Indonesia reaches large parts of the population, while coverage is related to socio-economic status and urban/rural location. The successes in Asia emphasize the need for multiple strategies and integration with other health and agricultural programs. PMID- 15806869 TI - Vegetarian diets. PMID- 15806870 TI - The contribution of vegetarian diets to human health. AB - Our knowledge is far from complete regarding the relationship between vegetarian diets and human health. However, scientific advances in the last decades have considerably changed the role that vegetarian diets may play in human nutrition. Components of a healthy vegetarian diet include a variety of vegetables, fruits, whole grain cereals, legumes and nuts. Numerous studies show important and quantifiable benefits of the different components of vegetarian diets, namely the reduction of risk for many chronic diseases and the increase in longevity. Such evidence is derived from the study of vegetarians as well as other populations. While meat intake has been related to increased risk for a variety of chronic diseases, an abundant consumption of vegetables, fruits, cereals, nuts, and legumes all have been independently related with a lower risk for several chronic degenerative diseases, such as ischemic heart disease, diabetes, obesity, and many cancers. Hence, whole foods of plant origin seem to be beneficial on their own merit for chronic disease prevention. This is possibly more certain than the detrimental effects of meats. Vegetarian diets, as any other diet pattern, have potential health risks, namely marginal intake of essential nutrients. However, from the public health viewpoint the health benefits of a well-planned vegetarian diet far outweigh the potential risks. PMID- 15806871 TI - Nutrition ecology: origin and definition. PMID- 15806872 TI - Promoting the nutritional and cultural benefits of traditional food systems of Indigenous People. PMID- 15806873 TI - Indigenous food and lost diversity. PMID- 15806874 TI - Possibilities and limitations of dietary therapy for inborn errors of metabolism. PMID- 15806875 TI - Genetics. Implications for nutrition. PMID- 15806876 TI - Nutritional status of Hungarian pregnant women. PMID- 15806877 TI - Nutrition of pregnant and lactating women. PMID- 15806878 TI - Infant and young child nutrition promotion strategies--the Malaysian experience. PMID- 15806879 TI - A new global strategy for infant and young child feeding. PMID- 15806880 TI - The WHO Multicentre Growth Reference Study: strategy for developing a new international growth reference. PMID- 15806881 TI - Complementary feeding strategies to improve child growth in developing countries. PMID- 15806883 TI - The relationship between nutrition and growth under various environmental conditions. PMID- 15806882 TI - Determinants of anaemia and iron deficiency in European infants at 12 months of age (Euro-Growth Study). PMID- 15806884 TI - Cardiovascular risks in childhood in Hungary. PMID- 15806885 TI - Global patterns of child health: the role of nutrition. PMID- 15806886 TI - New predictive equations for estimation of basal metabolic rates of adolescents: tropics versus temperate. AB - The energy and nutritional requirements of adolescents are influenced primarily by the growth spurt that occurs at puberty, and also by the need to maintain adequate levels of physical activity. Predictions of BMR have gained attention since the publication of the FAO/WHO/UNU (1985) expert consultation report, which adopted the principle of relying on energy expenditure rather than energy intake to derive requirement of individuals. While the report predicts BMR accurately in many individuals from temperate climate, they are said to be less accurate in predicting BMR in populations living in the tropics. The collation of worldwide data on basal metabolism indicated that, relative to adults, there was a paucity of data in other age groups including the adolescents. Although several BMR studies among children had been reported in the 90's, the data in normal weight children are almost exclusively from small control groups in obesity studies. Furthermore, we know little as to whether BMR of children differs in differing climatic conditions. This paper presents predictive equations for estimating BMR from a two-centre study, Bangi (Malaysia) and Oxford (UK) and to compare the results with the currently used predictive equations. PMID- 15806887 TI - Adolescent nutrition--a multidimensional challenge. PMID- 15806888 TI - The European Youth Heart Survey (EYHS): an international study that addresses the multi-dimensional issues of CVD risk factors. PMID- 15806889 TI - Aging, food patterns and disability. AB - To lay the groundwork for devising, improving and implementing strategies to prevent or delay the onset of disability in the elderly, a systematic literature review was conducted of longitudinal studies published between 1985 and 2001 that reported statistical significant associations between individual baseline nutritional risk factors and subsequent functional status in community-living older persons. The review revealed that nutrition has been neglected in past research. Not more than three studies fulfilled the methodological criteria (e.g. control for baseline functional status, control for confounding or attrition rate). The association between increased and decreased body mass index and functional decline was high. This and other available information regarding the importance of nutrition for the disablement process underline how important adequate intakes of energy and nutrients today are for the ability to manage activities of daily living tomorrow. Vulnerable groups of elderly people can be identified by questions on number of meals a day, food group intake and assessment of weight change. Policy makers and health care professionals should put a joined effort in optimising the food intake of these vulnerable elderly people. On one hand by stimulating physical activity to animate food intake and on the other hand by facilitating the availability of attractive nutritious foods. PMID- 15806890 TI - Age-fitness. How achievable with food? AB - To be fit as we age could be described as "Age fitness". Increasing evidence points to opportunities for greater wellness, health maintenance and reduction of the burden of disease in later life for a growing proportion of the community than previously envisaged. The scope of "age fitness" is social, mental, physiological and physical. We know a great deal about food cultures associated with successful aging--Greek, Okinawan, and Scandinavian, for example. Within these cultures, we are beginning to account for variances in health and survival by integrative scores of food intake and their validation. PMID- 15806891 TI - Antioxidants status and physical fitness in seniors aerobically trained and supplemented with a multivitamin drink. PMID- 15806892 TI - Dynamics of the nutrition transition and its implications for the developing world. PMID- 15806893 TI - Strategies for effective and sustainable national nutrition plans and policies. PMID- 15806894 TI - Trends in the burden of disease due to under- and over-nutrition in the developing world. PMID- 15806895 TI - Can the challenges of poverty, sustainable consumption and good health governance be addressed in an era of globalisation? PMID- 15806896 TI - Consequences of market economy on food and nutrition in Central and Eastern European countries. PMID- 15806897 TI - Cognition and undernutrition: evidence for vulnerable period. PMID- 15806898 TI - Functional foods and cognitive performance. PMID- 15806899 TI - Africa symposium--an introduction. PMID- 15806900 TI - Nutritional problems of Africa--the future of a continent: an overview. PMID- 15806901 TI - Multiple approaches to micronutrient deficiency in developing countries. PMID- 15806902 TI - Multiple micronutrients approach: the hope for improving micronutrient deficiencies in Africa. PMID- 15806903 TI - Control and prevention of micronutrient malnutrition. The way forward for speedy progress. PMID- 15806904 TI - The African nutrition leadership and Capacity Development Initiative. PMID- 15806905 TI - Why is malnutrition not declining in Africa? PMID- 15806906 TI - Focus on Africa: the state of nutrition and the challenges for the 21st century. PMID- 15806907 TI - Interventions to prevent low birthweight--a (South) African perspective. PMID- 15806908 TI - The future of animal experiments in nutrition research. AB - An overview is given on the legal framework in Europe for the use of experimental animals set by EU (European Union) Directive 86/609/EEC and on the activities of EU member states to implement this directive for regulatory testing in animals. Recent progress in implementing the 3Rs concept of Russel and Burch into regulatory guidelines of the EU are described. Moreover the validation of non animal tests to be used for regulatory purposes is outlined. From the regulatory and the animal welfare perspective the international harmonisation of test guidelines and the mutual acceptance of data are the way forward for chemical safety testing including food additives and contaminants. Examples are given for the replacement of diagnostic animal tests by analytical test methods, e.g. hormones, Clostridium botulinum toxins and neurotoxins from algae in shellfish. PMID- 15806909 TI - Validity of body composition methods across ethnic population groups. AB - Body composition methods can be classified into direct, indirect and doubly indirect methods. In vivo direct methods use neutron activation analysis to get information on body composition. Indirect methods rely on rules and constants derived from direct methods. Most basic research, especially the development of rules and models has been done in Caucasian subjects in Europe or USA. The critical use of more advanced body composition methodologies in various ethnic groups has shown that assumptions may differ between ethnic groups, an example being the assumption of constant density of the fat free mass. Indirect or predictive methods rely on statistical relationships between body parameters and components of body composition. Subcutaneous fat patterning differs among ethnic groups, and this may have consequences for the validity of body fat predicted from skinfold thickness. Relative leg length and relative arm length also differ between ethnic groups. As a result the body mass index (weight/height squared, BMI), often used as surrogate for body fat percent, and formulas based on bioelectrical impedance measurement show different validity among ethnic groups. Less information is available about the validity of indicators for body fat distribution. There are indications that the relationship between the amount of visceral adipose tissue and waist circumference or waist-hip circumference ratio also differs among ethnic groups. Ethnic differences in body composition rules and constants are important and challenging to investigate, especially in relation to overweight and obesity. PMID- 15806910 TI - Metabolically active components of fat free mass (FFM) and resting energy expenditure (REE) in humans. AB - Magnetic resonance imaging (MRI) can be used to assess metabolically active components of FFM. MRI-derived organ mass (OM) explains part of the interindividual variance in REE. It may also add to our understanding of malnutrition- and obesity-related variance in REE. There is need for (i) standardisation of MRI technology in body composition research, (ii) reference data on detailed body composition including also more recent autopsy data, (iii) reducing the number of assumptions in model-based predictions and (iv) a combination of imaging technologies with in vivo measurements of individual OM respiration (e.g. by positron emission tomography). PMID- 15806911 TI - Oxidative stress and antioxidant status of athletes and untrained persons. PMID- 15806912 TI - Functional assessment of nutritional status: lessons learned and future challenges. PMID- 15806913 TI - Why and how to use radioisotopes in human nutrition research. AB - Tracer methodology using radioisotopes in human nutrition research has been the object of unjustified limitations in its use. These limitations have encumbered research studies where micronutrient and macronutrient deficiencies are prevalent. Stable isotope methodologies are expensive, including the cost of the isotopes, equipment and its maintenance, and are often fraught with serious pitfalls. We have been developing safe radioisotopic methods to study the metabolism of micronutrients, especially iron and zinc in the developing world. These should allow scientists everywhere to conduct their own research without having to depend on facilities and laboratories in the industrial world, and thus enhance productive collaborative research. All radioisotopic research must be subjected to strict safety criteria, based on scientific evidence of risk, but should not be hampered by unscientific fears. Examples on the use of radio and stable isotopes to study iron and zinc absorptions are given including the radiation exposure and risk calculations. PMID- 15806914 TI - 2001 uses of stable isotopes in the assessment of nutrient status and metabolism. AB - Stable isotopes are nonradioactive and can be safely administered to humans; yet, because of the isotopic difference, can be distinguished from the unlabeled moiety and thus trace the nutrient uptake and elimination. Stable isotope applications include measurement of nutrient absorption, determination of nutrient body stores, tracing routes of nutrient metabolism, measuring nutrient fluxes through specific pathways, and measuring nutrient elimination. The ability to assess the dynamics of nutrient metabolism in vivo has been vital in the study of nutrient requirements, nutrient metabolism, mechanisms of nutrient homeostasis, and nutrient toxicity. Stable isotopes provide a window into human metabolism that is particularly valuable to the quantitative study of human nutrition. PMID- 15806915 TI - Nuclear techniques in nutrition and health: importance and applications in developing regions. AB - Nuclear and isotope methods in one form or another are regarded as essential tools for carrying out nutrition research besides enhancing sensitivity of nutrition monitoring techniques. They have been used extensively in industrialized countries to analyse human energy requirements, body composition including bone mineral density, and the metabolism of important nutrients such as protein, fat, vitamins and minerals. The information acquired has led to many improvements in nutrition and health. Currently, several strategic applications of isotopic techniques are being introduced in developing countries where they can benefit millions through monitoring improvement in nutritional status, and serve as specific indicators of broader social and economic advances. PMID- 15806916 TI - Using stable isotopes to assess the bioavailability of minerals in food fortification programs. AB - Food fortification with minerals is often undertaken without consideration of the bioavailability or nutrient-nutrient interactions related to its use. Stable isotopes provide a safe and accessible approach to providing direct evidence regarding these issues. Studies can be applied in all areas of the world and in all population groups. Mineral stable isotopes are safe and meet stringent ethical standards for use in children. Clinical studies in children allowing scientists, policy makers and the food industry to obtain data needed to understand how best to fortify foods to enhance the nutritional health of children of all ages. It is not necessary to have analytical mass spectrometry equipment available in each country where studies are performed. Rather, international collaborations can be developed to allow clinical studies to be performed in the field setting with analysis elsewhere if needed. The combination of isotope studies with field intervention trials may be beneficial. PMID- 15806917 TI - Transgenic potato plants expressing antimicrobial activity: establishment of pest control and environmental safety. PMID- 15806918 TI - Quality of plant-derived products. PMID- 15806919 TI - Contribution of animal food in the form of meat to human nutrition. PMID- 15806920 TI - What is pork and poultry meat quality? PMID- 15806921 TI - Risk as a social phenomenon. AB - What do nuclear power, smoking, and eating beef have in common? The answer is multifaceted. They all share aspects of mass production and consumption, and therefore consumer confidence comes into play in evaluating each of the products' merits. They all produce some level of anxiety--be it in response to adverse media coverage, or the successes of adversarial interest groups in publicizing those products' weaknesses--among consumers who wonder what if any level of the product should be consumed. Finally, nuclear power, smoking, and eating beef all share risk; that is, the perceived and real detriments of producing and consuming those products. Whether or not the associated risks--from increased cancer to increased heart disease--are real is beside the point. When talking about risk, scientific education must deal with the perception of risk as much as its reality. Risk is a socially defined phenomenon, and as such, the successful scientists/communicator must understand the social evaluations of it. While many scientists take a rational view of risk evaluation and consumers often make rational decisions about technological and scientific risk based upon the costs and benefits of that technology, this approach is problematic for three reasons. First, the nature of modern risks has changed in that risk is complex, incomprehensible and uncontrollable to the average person. Second, the rational approach assumes that all costs and benefits of a technology, both real and potential, both current and future, can be known and therefore rationally evaluated. This is not the case. Third, evidence from Europe indicates that trust in information sources about risk has fallen, and thus classic scientific education campaigns that rely upon increasing public knowledge and acceptance of technological and scientific risk are problematic. Because of these factors we argue that effective scientific response must begin to understand the social components of risk if campaigns to shape public perception are to have any success. The report concludes that public perceptions concerning risk are driven more by social and moral debates than by scientific or economic debates (340 words). PMID- 15806922 TI - New recommendations for the energy and nutrient supply of dairy cows and heifers in Germany. AB - In summary, GfE (2001) give not only a view on energy and nutrient requirements, and the official recommendations for supply in Germany established by the Society of Nutrition Physiology, but also a comprehensive literature review on the particular issues. Some gaps of knowledge on requirements of dairy cows and heifers were identified and the need of further research was determined. The derivation of the new recommendations under consideration of physiological aspects has been demonstrated in the paper. PMID- 15806923 TI - Ruminant nutrition in developing countries. PMID- 15806924 TI - Assessment of novel foods in animal nutrition. AB - Composition of feeds from GMO crops were determined as well as digestion and feeding experiments were carried out with broilers (Bt-corn), layers (Bt-Corn, Pat-corn), pigs (Bt-corn, Pat-sugar beet, soybeans), sheep (Bt-corn silage, Pat corn silage), growing bulls (Bt-corn silage) and fistulated cows (Bt-corn silage). Up to now, no significant differences in nutritional value between feeds from isogenic and transgenic plants of the first generation were observed. The so called substantial equivalence, but also the nutritional equivalence of transgenic hybrids could be demonstrated. Recombinant plant DNA constructs were not detected in animal tissues samples. In the future, long term feeding experiments for nutritional assessment of novel feeds should be combined with risk assessment studies. Proposal for discussion has been submitted. PMID- 15806925 TI - Impact of processing on nutritional quality of marine food items. AB - During the last two decades it has been established that Greenland Eskimos living on their traditional diet, have a lower incidence of coronary heart disease (CHD) than when living in Denmark on a western diet. These findings have been attributed to their diet, particularly the high amounts n-3 polyunsaturated fatty acids. The Eskimo diet consists mainly of meat and blubber of seal and whale and relatively small amounts of fish. Another aspect of the Eskimo diet, still not fully explored, is that the Eskimos consume the bulk of their food raw or dried, seldom boiled or exposed to excessive heat. The main task of modern processes is to make edible and stable products. Removal of molecules that cause off-flavours or taste to improve sensory attributes may, for instance, destroy potent antioxidants. Modern meal preparing techniques may also lower the content of biologically active components. The objective of our experiments has been, by mimicking in part the traditional Eskimo diet, to explore the beneficial effects of raw food items on parameters related to development of CHD. Reduced tendency of developing arteriosclerosis has been related to the lower reactivity of platelets and less production of proinflammatory products, e.g. cytokines, prostaglandins and leukotrienes. In our study, healthy volunteers ingested raw or heat processed marine materials (smoked versus heat processed salmon muscle, cold pressed versus refined marine oils). PMID- 15806926 TI - Current issues in fish and shellfish nutrition. PMID- 15806927 TI - Losses of natural antioxidants and vitamins during deep-fat frying. PMID- 15806928 TI - Frying oil discarding: polar content vs. oligomer content determinations. AB - The debate on deep-fat frying is primarily focused on the point at which any oil used for frying should be discarded. Polar content (PC) and triacylglycerol oligomer content (TOC) determinations constitute the basis of legislation for oil discarding in some European countries. Most of them have set a maximum level of 25% PC while others have established a PC cut-point between 20% and 27%. The maximum TOC have been set at 10% while other countries permit 16%. The aim of the present study was to study the relationship between these two measurements and to open some debate on whether PC or TOC is preferred for oil discarding assessment. Monounsaturated oils (olive oil, extra virgin olive oil, high oleic acid sunflower oil), polyunsaturated oils (sunflower oils and a blend of olive oil and sunflower oil), and saturated oils (palm olein) used in frying of different foods under domestic controlled conditions were employed. Further application of HPSEC to the isolated PC to measure TOC was preferred to the direct evaluation of TOC. Although PC and TOC were always very high correlated (p < 0.001), the 25% PC corresponds to a 10% TOC in palm olein but to 15% in sunflower oil. These findings suggest the need to unify criteria for oil discarding. Taking into account the potential toxicity of oligomers and also that some compounds defined as polar components are not necessarily altered (e.g. diacylglycerols, free fatty acid), it can be suggested that TOC gives more precise information about the alteration of the oil and its potential toxicity than the PC. PMID- 15806929 TI - Ascorbyl palmitate and its synergism to tocopherols. PMID- 15806930 TI - Effect of food component changes during frying on the nutrition value of fried food. PMID- 15806931 TI - Nutritional consequences of food processing. AB - A variety of methods are used to process foods: if they are not edible, to render them so; to permit storage; to alter texture and flavor; to destroy microorganisms and other toxins. These methods include heating (baking, cooking, frying, microwaving), freezing, and high pH. It is a paradox of nature that the processing of foods can improve, nutrition, quality, and safety; yet, occasionally these processing alternatives can lead to the formation of anti nutritional and toxic compounds. These multi-faceted consequences of food processing result from molecular interactions among nutrients and with other food ingredients, both natural and added. This paper outlines the following aspects of processing-induced formation of novel food ingredients and the resulting consequences for nutrition: protein-polyphenol and protein-carbohydrate enzymatic and non-enzymatic browning reactions; formation of heterocyclic amines in meat; inactivation of soybean inhibitors of digestive enzymes; formation of lysinoalanine and D-amino acids in food proteins; and the stability of phenolic compounds to high pH. Possible approaches to prevent the formation of deleterious food ingredients are also addressed. PMID- 15806932 TI - Maillard reaction products: uptake, metabolic transit and selected parameters of biopotency and safety. PMID- 15806933 TI - New aspects of the functionalities of probiotics. PMID- 15806934 TI - Discussion points on the functionalities of probiotics. PMID- 15806935 TI - Importance, impact evaluation, quality assurance and safety of food fortification in developed countries. PMID- 15806936 TI - Food fortification: issues on quality assurance and impact evaluation in developing countries. AB - Quality assurance and impact evaluation are essential components of a food fortification program and should be integrated in the fortification process. Quality assurance will ensure that the micronutrient meant to be delivered is indeed reaching the target population at the correct level. Impact evaluation will determine the effectiveness of food fortification as a strategy in controlling micronutrient deficiency and enable program planners to make decisions on the future of the program. In developing countries, both quality assurance and impact evaluation are often constrained not only by inadequacy of facilities as well as financial and manpower resources, but by unclear definition of objectives and inappropriate design. It is therefore necessary to consider the target audience for the quality assurance monitoring and impact evaluation in order to clearly define their objectives and in turn suit the design to these objectives, at the same time as the limitations in financial and manpower resources are considered. PMID- 15806937 TI - Antioxidant supplementation and risk of chronic disease. PMID- 15806938 TI - Supplements and food fortification--when and where? PMID- 15806939 TI - New food technologies and processes and their impact on nutrition. AB - An evaluation of new technologies already applied or still under investigation has shown that those procedures allow a mild treatment mostly of liquid food materials. If also Osmotic Processing, Ionising Irradiation, Carbon-Dioxide High Pressure-Gas treatment, Ultra-Sound treatment and Gas-Plasma treatment are considered a large array of new processes is available for processing of food with minimum impact on compounds of nutritional relevance. Those processes must however be operated under very special conditions in order to comply with hygienic standards. Since all this processes require very sophisticated equipment it is too early at the moment to predict which one of the New Technologies will survive the fierce competition with the classic and well established processes and which do not fall under the Novel Food Regulation of the EU. It must be stated however that the pressure which did arise from the New Technologies has initiated many developments in the area of the classical technologies which had at the very end an impact also on the quality of products produced with those procedures. PMID- 15806940 TI - Finding optimal photosensitisers for the decontamination of foods by the photodynamic effect. AB - A new method for sanitation based on the photodynamic effect is introduced. Photosensitisers are compounds that generate cytotoxic oxygen species and free radicals after excitation with appropriate visible light. The cell killing efficacy is due to their chemical structure, to the actual environment and therefore also to their degree of aggregation. As has been shown, photosensitisers bound to the cell wall or incorporated into the cells are very effective cytotoxic agents after illumination. Even singlet oxygen generated in the gas phase surrounding the micro-organisms effectively kills cells. Especially membrane components are the targets of the photodynamic effect. Membrane disintegration is observed soon after illumination. The food contaminants under investigation are Bacillus subtilis, Bacillus cereus, forming endospores, Staphylococcus aureus, forming a wide range of agressins, exotoxins and enterotoxins, all gram positive micro-organisms, Escherichia coli, a hygiene indicator, and the yeasts Saccharomyces cerevisiae, Kloeckera javanica, Rhodotorula mucilaginosa, a blue and a pink pigmented isolate from foods. We investigated the capacity of several photosensitisers to induce the phototoxic effect towards those micro-organisms following excitation by visible light. PMID- 15806941 TI - Non-thermal food preservation and its potential in meat processing. PMID- 15806942 TI - Nutritional challenges of the 21st century. PMID- 15806943 TI - Food security in a world without borders. AB - The alleviation of poverty and the eradication of hunger and malnutrition are within reach. Considerable progress has been made over the past thirty years in reducing the numbers of the hungry, and projections over the next thirty years suggest that this progress will continue. The majority of developing countries have participated in this progress and have improved nutrition but there are significant regional differences. The current challenge is to build upon and accelerate the progress already made. Everyone is involved in this struggle against hunger and malnutrition, and to achieve these goals, global partnerships to enhance co-operation and co-ordination are being strengthened. Successful country experiences for improving food security and nutrition have demonstrated the importance of peace, political stability, and stable economic growth. There is also the need for increased foreign investment and increased ODA, particularly for African agriculture, debt relief, the better integration of LDCs in the global economy, their more secure access to markets and more equitable terms of trade, enhanced South-South co-operation, training and research and new high yield and drought-resistant crop varieties. While all these have been commented on and recognised before, we must close the credibility gap in our political will by honouring our existing commitments for providing tangible benefits at the local level. For globalisation, the key issue is how the aggregate benefits of globalisation will be distributed and how to translate this into better nutrition. The LDCs, which are the most vulnerable and disadvantaged members of the international community, should be at the centre of this drive towards food security and take genuine ownership of policies, initiatives and activities to improve development. At the same time we need to succeed in making the developed world more aware of and responsive to the conditions of the LDCs. The most effective way to improve nutrition is through community nutrition programmes that encourage the full participation and co-operation of the entire community, maximise the utilisation of local resources, grasp the benefits of new technologies for productivity gains, involve multiple sectors and engage strong political commitment. The international community will be judged by its treatment of its most vulnerable members. The international community has repeatedly declared that it is dedicated to the eradication of poverty. Eliminating hunger and malnutrition is a vital first step. The political will to fight hunger and a firm commitment to invest in agriculture' and rural development are critical elements in any effort to achieve sustainable alleviation of hunger and poverty. This meeting and the WFS:FYL provide the additional impetus to meet the challenge of achieving food and nutritional security in a world without borders and free from hunger and malnutrition. PMID- 15806944 TI - The role of non-government organizations in promoting nutrition and public health. PMID- 15806946 TI - No longer off the menu: the welcome re-emergence of food on the nutrition agenda. PMID- 15806945 TI - Food security and nutrition--a household perspective. PMID- 15806947 TI - Assessing the impact of large scale nutrition interventions. PMID- 15806948 TI - Evaluation challenges with respect to large-scale nutrition intervention programs. PMID- 15806949 TI - Nutrition security and crisis prevention: can nutritional science contribute to crisis prevention? PMID- 15806950 TI - Food aid in emergencies and public health nutrition. AB - HUNGER LEAVES SCARS: The visible kind may be born by the survivors of famine. Less visible, but all the more damaging, are the long-term effects of hunger that run through families through generations. Hunger passed from mother to child represents a ruinous inheritance. It marks a cycle of hunger that transcends generations, unless the cycle is broken. Food aid provided at crucial times in the lives of women and infants represents an investment for future. Thus, in contrast to former conceptions of food aid as an exclusively life-saving vehicle, modern aims of food aid also include preventing increases in the prevalence of malnutrition and asset depletion. Mass migration and food shortages have been responsible for most deaths following civil conflicts around the world. The most visible form of migration occurs when people cross international borders. The reasons for the flight of refugees and internally displaced persons are generally same; war, civil strife, and persecution. "NUTRITIONAL GATEWAYS": Finally, the importance of timely and sustained delivery of adequate food aid adequate in quality and quantity to people in dire need during the emergency is paramount. Food aid is the most direct means for conveying nutritional benefits: the time frame is often limited, sustainability is not an issue. However, in the case of drought victims, refugees or displaced people, the nutritional situation and the actions needed are more complex. In many situations people arrive are often in very bad state. While high prevalence of malnutrition is associated with inadequate food rations, in some situation malnutrition developed primarily because of the high incidence of diarrhoeal diseases. The synergism between high malnutrition and increased incidence of communicable diseases explains much of excess mortality seen in refugee and displaced persons. PMID- 15806951 TI - Bovine spongiform encephalopathy (BSE): the importance of the food and feed chain. PMID- 15806952 TI - Water quality and health in the new millennium: the role of the World Health Organization Guidelines for Drinking-Water Quality. AB - In this report the role of the WHO Guidelines for Drinking Water Quality in promoting safe drinking water for the world's population is briefly described. The guidelines are being revised in a third edition to emphasize an integrated approach to water quality assessment and management from source to consumer. The forthcoming guidelines will: be risk-based and quantitative, emphasize quality protection and prevention of contamination, be proactive and participatory, and address the needs of those in developing countries who have no access to piped community water supplies. The guidelines emphasize the maintenance of microbial quality to prevent waterborne infectious disease as an essential goal. In addition, they address protection from chemical toxicants and other contaminants of public health concern. The forthcoming 3rd edition of the WHO GDWQ intend to be responsive to the under-served in developing countries by inclusion of non piped supplies and addressing practical systems for their collection, treatment and storage at household level to provide safe water. Beyond the inclusion of these and possibly additional household water collection, treatment and storage systems, what is needed is to achieve their widespread use is an education and dissemination campaign that promotes and explains them and their benefits. Such a communication and marketing campaign is best done by including as many different sectors and stakeholders as possible in the process. It will be important to acknowledge that safe water is one of essential components or needs for healthy living, along with adequate sanitation and proper nutrition. Together, these are the essential health needs to be met in the developing and the developed world. All three contribute to reduced disease and increased health, and the lack of one can degrade the beneficial impact of the others. The importance of safe water, sanitation and nutrition to human health and well-being can be stated no better than it was by United Nations Secretary General Kofi Annan in his statement on "Freedom from Want" in the Millennium Report, 03/04/00. "How can we call human beings free and equal in dignity when over a billion of them are struggling to survive on less than one dollar a day, without safe drinking water, and when half of all humanity lacks adequate sanitation? Some of us are worrying about whether the stock market will crash, or struggling to master our latest computer, while more than half our fellow men and women have much more basic worries, such as where their children's next meal is coming from." PMID- 15806953 TI - Risk reduction of endocrine disruption by antioxidants. PMID- 15806954 TI - Food safety and risk assessment: naturally occurring potential toxicants and anti nutritive compounds in plant foods. AB - The centralisation and globalisation of the world food supply presents opportunities for massive epidemics of food borne illness. Food processing generally decreases exposure to naturally occurring toxicants. A significant number of deaths are caused in both developed and developing countries from the consumption of naturally occurring toxicants and this exceeds the numbers attributable to artificial food contaminants and pollutants. The risk posed by naturally occurring toxicants is greater in developing countries where food choice and facilities for food processing are limited. The main risk in developed countries is from food important from areas where food safety controls are inadequate. Toxins derived from marine algae present a specific threat to health especially as seafood is being traded from areas where poisoning by algal toxins is known to occur. A systematic approach to food safety using HACCP methodology and identity preservation of the food are crucial in the prevention of food related illness. There is also a need for international agreed standards for tolerable levels of naturally occurring toxicants in foodstuffs but these should derived not be set unreasonably low as this would threaten food security in developing countries. The occurrence of a number of fatalities in developing countries among people consuming herbal teas and dietary supplements underscores the need to assess the safety of these products before they are placed on the market. PMID- 15806955 TI - Mycotoxins: natural toxins of agricultural and health significance. PMID- 15806956 TI - Actions of the antibiotic mycotoxins beauvericin and enniatin on mammalian tissue. PMID- 15806957 TI - Microbiological criteria for safe food. PMID- 15806958 TI - New approaches for the fast detection of microorganisms in food and water using enzyme detection method (EDM). PMID- 15806959 TI - Food control systems. PMID- 15806960 TI - [Progress on research of tissue culture of Siraitia grosvenorii]. AB - In this paper we reviewed the development of tissue culture, current situations of virus-free plantlets industrialization and the way to deal with the situations, application prospects of Siraitia grosvenorii so as to give some advice for its further study and application. PMID- 15806961 TI - [Studies on the botanical sources and DNA molecular identification of Herba Asari based on ITS sequence]. AB - OBJECTIVE: To explore the genetic basis of using three species of Asarum as Herba Asari to determine the taxonomic positions of Asarum heterotropoides and A. siebodii; and to apply DNA molecular analysis as a tool for identification of Herba Asari. METHOD: PCR, purification, sequence analysis were prerformed. RESULT: MS sequences of the three Asarum species were obtained. 3 botanical sources of Herba Asari are closely clustered together on the topology tree; one inner branch is composed of A. heterotropoides and A. sieboldii, whereas another branch contains A. sieboldii. Their ITS sequences are different. CONCLUSION: Three plant species of Herba Asari are closely related, and there are genetic reasons that they are used as the sources of the same medicine. The classification placement of A. sieboldii is not certain. The differences of ITS sequences of the botanical sources of Herba Asari can be used as a means of identification. PMID- 15806962 TI - [Analysis on the geologic background and physicochemical properties of soil for the cultivation of Panax notoginseng in Yunnan province]. AB - OBJECTIVE: To study the relationship between the geo-authentic and geologic background system (GBS) and physicochemical properties of soil for the cultivation of Panax notoginseng. METHOD: The physicochemical properties of soil were analyzed, the geologic background system between producing areas of Yunnan and Guanxi of P. notogiseng was compared. RESULT: The distribution of P. notoginseng was restricted by GBS, The yellow-red soil of fragmentary rock mixed with carbonate seems to be most suitable for the growth. CONCLUSION: The most suitable soil for growing P. notoginseng is the neutral or weakly acidic soil with loam clay, particularly that with low base saturation percentage. PMID- 15806963 TI - [Influence of process methods on contents of chemical component Radix Polygoni Multiflori]. AB - Different methods of processing Radix Polygoni Multiflori and the change of chemical components including emodin, physcion, 2,3,5,4'-stilbene glucoside, and tannin were investigated. The dried roots of polygoni were processed in different ways, such as, steamed with black bean sauce, steamed with water, stewed with black bean sauce, and stewed with black bean sauce at high pressure, for h and 36 h, respectively. The process stewing with black bean sauce was also investigated for 12, 24, 36 h, and 48 h, respectively. The results indicated that 2,3,5,4' stilbene glucoside and total free anthraquinones varied in different processes. In the process of black bean sauce stewing, the content of 2,3,5,4'-stilbene glucoside decreased with processing time, reached 17% of original at 48 h. The content of tannin and combined and free anthraquinones also decreased with processing time. It was concluded, therefore, that the processing of Radix Polygoni Multiflori should be standardized to avoid significant variation of chemical contents. PMID- 15806964 TI - [Enhancement of thrombolytic activities of Carthamus tinctorius processed with fermentation with a bacillus sp. C2-13]. AB - A processing method to enhance thrombolytic effect of Carthamus tinctorius using a fermentation technology with bacillus sp. C2-13 was investigated. The fibrinolysis and anticoagulation activity of thrombolytic extracts from an optimized fermentation process was studied using a carrageenan induced mice model. The fermented extracts resulted in significantly better thrombolytic activity, suggesting that the process was promising for use in the study and preparation of nature medicines. PMID- 15806965 TI - [Study on the encapsulation technique of high purity gamma-linolenic acid, part 1 -saponification reaction and saponification value]. AB - OBJECTIVE: To measure the saponification value and fatty acid formation of evening primrose oil, to study the effects of pH value on production yield and fatty acid formation during the saponification reaction, and to provide rationales for the selection of raw material, the enhancement of production yield of saponification, and the encapsulation of gamma-linolenic acid with urea. METHOD: To measure fatty acid's formation with gas chromatographic method and to measure the saponification value. RESULT: The content of gamma-linolenic acid is 7%-10% in evening primrose oil. The content of gamma-linolenic acid is inversely correlated with that of unsaturated fatty acid. The saponification value, the amount of KOH for saponification of evening primrose oil, and the pH value for subsequent isolations of oils are determined. From the measurement of fatty acids of evening primrose oil in two different cultivation locations, the content of gamma-linolenic acid is determined to be 7%-10%, unsaturated oils account for 90%. CONCLUSION: The saponification value of evening primrose oil is between 180 200, pH value of isolated oil is 1.5-2.0 after saponification reaction. Fatty acids mainly include palmitic acid, stearic acid, oleic acid, linolic acid and gamma-linolenic acid. PMID- 15806966 TI - [Chemical constituents from marine alga Chaetomorpha basiretorsa]. AB - OBJECTIVE: To investigate the chemical constituents of marine alga Chaetomorpha basiretorsa. METHOD: Compounds were isolated by normal phase silica gel and Sephadex LH-20 gel colum chromatography, reverse phase MPLC, reverse phase HPLC and recrystallization. Their structures were elucidated by spectroscopic methods including MS, IR, NMR, and X-ray crystalography. Cytotoxicity of the compounds were screened by using standard MTT method. RESULT: Nine compounds were isolated from C. basiretorsa and their structures were identified as N-phenyl-2 naphthalenamine( I ), dibutyl phthalate( II ), diisobutyl phthalate( III ), 1 phenyl-ethane-1, 2-diol( IV ), 2-hydrox-gamma-benzaldehyde( V ), diethyleneglycol monobenzoate( VI ), uracil( VII ), thymine( VIII ) and thymidine( IX ). CONCLUSION: All these compounds were obtained from this genus for the first time, N-phenyl-2-naphthalenamine and diethyleneglycol monobenzoate were first reported from the marine organisms. Compound I and VII showed moderate activity against KB cell(IC50 10.15 microg x mL(-1) for I and 3.79 microg x mL(-1) for VII ) and MCF 7 cell(IC50 3.24 microg x mL(-1) for VII). PMID- 15806967 TI - [Studies on constituents from the fermentation of Alternalia sp]. AB - OBJECTIVE: To study the metabolites of marine fungus Alternalia sp. METHOD: Compounds were separated by column chromatography and their structures were elucidated by means of chemical and spectral analysis. RESULT: Six compounds were isolated from the ethyl acetate and n-butanol extracts of the fermentation of marine fungus Alternalia sp. Their structures were elucidated as p-benzyloxy phenol ( I ), p-hydroxy phenyl ethylamine( II ), 3-hydroxymethyl-8-hydroxyl pyrrolopiperazine-2, 5-dione ( III ), 3-isobutyl-6-secbutyl-piperazine-2, 5-dione (IV), 5alpha, 8alpha-epidioxy-ergosta-6, 22-diene-3beta-ol (V), 3beta-hydroxxy cholesta-5-ene (VI). CONCLUSION: Compounds I , II, III, IV have the activity of inducing morphological deformation of mycelia germinated from conidia of Pyricularia oryzae. Compounds I , II , III were isolated from the genus Alternalia for the first time. PMID- 15806968 TI - [Phenanthrene constituents from rhizome of Arundina graminifolia]. AB - OBJECTIVE: To isolate and elucidate the constituents from rhizome of Arundina graminifolia. METHOD: Theconstituents were extracted with 95% alcohol and isolated by chromatography on silica gel, Sephedax LH-20. The structures were determined by UV, IR, NMR and MS spectral analysis. RESULT: Five phenanthrene constituents were identified as 7-hydroxy-2, 4-dimethoxy-9, 10 dihydrophenanthrene( I ), 4, 7-dihydroxy-2-methoxy-9, 10-dihydrophenanthrene ( II ), 2, 7-dihydroxy-4-methoxy-9, 10-dihydrophenanthrene ( III ), 7-hydroxy-2 methoxyphenanthrene-1,4-dione ( IV ), 7-hydroxy-2-methoxy-9, 10 dihydrophenanthrene-1,4-dione (V), respectively. CONCLUSION: All compounds were isolated from rhizome of A. graminifolia for the first time. PMID- 15806969 TI - [The inhibition of 20(R)-ginsenoside Rg3 on the expressions of angiogenesis factors proteins in human lung adenocarcinoma cell line A549 and HUVEC304 cell]. AB - OBJECTIVE: To study the effect of 20( R)-ginsenoside Rg3 on the expressions of angiogenesis factors proteins (VEGF,bFGF, MMP-2) in human lung adenocarcinoma cell line A549 and HUVEC304 cell. METHOD: The cell lines of A549 and HUVEC304 were cultured with 20(R)- Rg3. The gray scale and positive rate of VEGF, bFGF, MMP-2 were detected by immunohistochemistry. The differential expressions of genes were studied by DNA microarray. RESULT: The positive rate of VEGF protein in A549 cell decreased significantly as compared with the control group ( P = 0.03). The gray scales of VEGF, Flt, KDT proteins in both A549 cell lines and HUVEC 304 cell lines decreased ( P = 0.05). Gray scale of MMP-2 also decreased in A549 cell lines. The result of differential expressions of genes of A549 cell lines showed that 14 genes were down-regulated and 10 genes were up-regulated. CONCLUSION: The Chinese materia medica of 20( R)-Rg3 can inhibit the expression of angiogenesis factors proteins via several target genes in both tumour cell and vascular endothelial cell. PMID- 15806970 TI - [Suppressive effects of GTW treatment on mesangial lesions in experimental irreversible glomerulosclerosis]. AB - OBJECTIVE: To examine suppressive effects of multi-glycoside of Tripterygium wilfordii Hook. f. (GTW)on mesangial injury induced by two-injecti on of anti Thy1. 1 monoclonal antibody(mAb) 1-22-3 in vitro. METHOD: We established the irreversible model of glomerulosclerosis with anti-Thy1. 1 mAb 1-22-3. After 42 days of oral treatment with GTW (50 mg x kg(-1) BW)and vehicle (distilled water), to observe effects of GTW on proteinuria, renal function, mesangial morphological change, and mRNA expressions of collagen type I and TGF-beta by light microscope (LM), immunofluorescence (IF), and Reverse Transcription Polymerase Chain Reaction (RT-PCR). RESULT: GTW ameliorated proteinuria (from day24 to day 42) and mesangial proliferation [total cell number, GTW group 65.67+/-3.43 vs. control group 87.02+/-2.41, P < 0.05; matrix expansion, GTW group 1.20+/-0.06 vs. control group 2.77+/-0.23, P < 0.05; alpha-smooth muscle actin(alpha-SMA) expression, GTW group 1.75+/-0.33 vs. control group 2.62+/-0.15, P < 0.05; collagen type I expression, GTW group 1.68+/-0.31 vs. control group 2.06+/-0.24, P < 0.05], moreover, significantly reduced the glomerular expression of mRNA for collagen type 1(53.5% to the control group, P < 0.05)and TGF-beta(14.7% to the control group, P < 0.05)on day 42day. CONCLUSION: GTW can not only decrease proteinuria, but also ameliorate mesangial alterations probably by the reduction of cytokines. GTW may be a promising agent for the prevention of progressive and irreversible glomerulosclerosis. PMID- 15806971 TI - [Study of huchang qingfei pellets on immune function in rats infected with Mycoplasma pneumoniae]. AB - OBJECTIVE: To study the effects and mechanism of Huchang Qingfei pellets on immune function in rats infected with mycoplasma pneumoniae. METHOD: A rat model of mycoplasmal pneumonia (MP) was developed in repeated intranasal infectious routes and then the humoral and cellular immunocompetences were detected by radioimmunoassay, immune-turbidimetry and flow cytometry. RESULT: The levels of serum IgG,IgM and IL-2, IL-6 were enhinced obviously, the complement C3 and TNF alpha were decreased and the ratio of CD4+ /CD8+ was improved significantly in the Huchang groups as compared with MP model group. CONCLUSION: Huchang Qingfei pellets can reinforce immune function via preventing both cellular and humoral immunity from depression in the rats with MP. PMID- 15806972 TI - [Study of the effect of crocin on rat experimental hyperlipemia and the underlying mechanisms]. AB - OBJECTIVE: To study the effect of crocin on rat experimental hyperlipemia and its mechanisms. METHOD: Hyperlipemia model was established by feeding heavy cholesterol for 2 months and the effect of crocin on blood lipid in experimental hyperlipemia rats was observed. Aortic smooth muscle cells were cultured in different culture media and proliferation was measured by MTT assay. Western blotting was used to detect the effect of crocin on phosphorylation of p38 MAPK. RESULT: Crocin not only decreased greatly the content of cholesterol, triglyceride and density lipoprotein in blood, but also increased the content of high density lipoprotein. In addition, the proliferation of smooth muscle cells and the activation of p38MAPK were inhibited by Crocin. CONCLUSION: Crocin prevents atherosclerosis in hyperlipemia, which may be mediated by the inhibition of both proliferation of smooth muscle cells and activation of p38MAPK. PMID- 15806973 TI - [Effects of jiantai liquid on the expression of estrogen/progesterone receptors in embryo implantation dysfunction mice endometrium]. AB - OBJECTIVE: To explore the mechanism of Jiantai liquid on the endometrium development of embryo implantation dysfunction mice. METHOD: The model of embryo implantation dysfunction mice was induced by mifepristone and treated by Jiantai liquid. All animals were sacrificed on day 8 of pregnancy. Estradiol and progesterone concentrations in serum and endometrium tissue homogenates were measured by radioimmunoassay method, the endometial expressions of estrogen receptor (ER)and progesterone receptor (PR)assessed by immunohistochemical SP method. RESULT: There were no significantly differences in the estradiol and progesterone concentrations in serum and uterus tissue homogenates among three groups( P > 0.05). Absorbency and area rate of ER, PR in model group' s gland and stroma were higher than those in model group(P < 0.05), which was similar with the control group( P > 0.05). CONCLUSION: Jiantai liquid increase the implantation rate and improve the endometrial development by increasing the expressions of ER, PR in endometrium of embryo implantation dysfunction PMID- 15806974 TI - [The renoprotect effect of shenhua recipe on 5/6 renal ablation rats]. AB - OBJECTIVE: To investigate the reno protective effect of Shenhua recipe on the experimental model of 5/6 renal ablation. METHOD: 5/6 renal ablation rats were underlying this experiment. They were administered Shenhua, irbesartan respectively by gavage during 12 weeks. Body weight, systolic blood pressure, proteinuria, Scr, BUN, total protein, albumin, Glycero and cholesterol were measured. Histologic glomenular and tubulointerstitial damage scores were measured at 12 weeks. RESULT: The treated groups showed significantly less histologic glomerular and tubulointerstitial damage scores at 12 weeks. The plasma albumin were higher ( P < 0.05), urine protein excretion rates, serum cholesterol and creatinine were lower than in nontreated group, but arterial blood pressure was not significantly different in the three Shenhua treated groups compared with nontreated group. CONCLUSION: Shenhua can retard the progression of chronic renal injury in the 5/6 renal ablation without changes in systolic blood pressure. PMID- 15806975 TI - [Clinical efficacy and T-lymphocyte subset, serum interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-2(IL-2) levels on treatment of chronic aplastic anemia patients by shenfu injection combined with stanozol and cyclosporin A]. AB - OBJECTIVE: To observe the effect of Shenfu injection (SFI) and influence on T lymphocyte subset, serum level of interferon-gamma(IFN-gamma), tumor necrosis factor-alpha(TNF-alpha), interleukin-2(IL-2) in patients with chronic aplastic anemia (CAA) based on treating with stanozol and cyclosporin A. METHOD: 60 patients with CAA were randomly divided into two groups, 30 patients in the SFI group were treated with SFI (100 mL which contains Ginsenoside 0.8 mg x mL(-1) and aconitine 1.8 microg x mL(-1) by adding it in 500 mL of 5% glucose every day) plus stanozol and cyclosporin A and 30 patients in the control group treated with slanozol and cyclosporin A alone for 2 months. The clinical efficacy was observed. The change of T-lymphocyte subset analyzed by flow cytometry and the levels of serum IFN-gamma, TNF-alpha, IL-2 measured with ELISA method were also observed before and after treatment. RESULT: After treatment, the total effective rate of the SFI group was higher than that in the control group, but it did not showing significant difference. The CD4/CD8 levels were significantly increased (1.76+/-0.49, P< 0.01) and CD8 levels were significantly lowered (22.57+/-6.30, P < 0.01) in the SFI group after treatment. Serum levels of lFN-gamma, TNF-alpha and IL-2 were lower in both groups, and the level of TNF-alpha and IL-2 in the SFI group (0.710+/-0.213) ng x L(-1) and (0.639+/-0.247) ng x L(-1) was significantly lowered than that in the control group (P < 0.05, P < 0.01). CONCLUSION: SFI might believe the hemopoietic inhibition so as to promote the recovery of hemopoietic function through improving the T-lymphocyte subset and reducing the release of hemopoietic negative regulatory factors such as IFN gamma, TNF-alpha and IL-2. PMID- 15806976 TI - [Studies on the anticancer effects of total alkaloid from Viscum coloratum]. AB - OBJECTIVE: To study the anticancer effects of total alkaloid from Viscum coloratum in vivo and vitro. METHOD: In vitro, MTT assays were used t o measure the inhibitory effect. Cells at period of logarithmic growth were incubated for 24 hours. Then total alkaloid of various concentrations were added. 24 hours later, supernatant was removed and MTT was added. 4 hours after that, DMSO was added, then 30 minutes later, A value was measured. In vivo, suspension of carcinoma cells was implanted in the mice's limbs subcutaneously, 0.2mL each. 24 hours later, the mice were grouped randomly. Fed by total alkaloid continuously for 7 days, the mice were sacrificed. The tumors were weighed and calculated the inhibitory rates. RESULT: In vitro, it shows that total alkaloid has prominent inhibitory effect on the growth of carcinoma cells. In vivo, it shows that total alkaloid can inhibit the growth of tumors and prolong the survival days of the mice bearing tumors. CONCLUSION: Total alkaloid from Viscum coloratum has the activities of anticancer. PMID- 15806977 TI - Effects of tea constituents on cell cycle progression of human leukemia U937 cells. AB - Tea and tea constituents are known to induce apoptosis in a variety of cancerous cells, suggesting their beneficial effects as chemopreventive agents. Previous studies have shown that low molecular weight constituent catechins and high molecular weight fractions of tea have the apoptosis-inducing activity, but that their action mechanisms may be different. Since cell cycle arrest is known to be one of the underlying mechanisms of apoptosis, we examined the effects of these tea constituents on cell cycle progression of human leukemia U937 cells. The results showed that the high molecular weight fractions of green tea and black tea caused G2/M arrest associated with up-regulation of p21/Waf1, but that epigallocatechin gallate, a major component of green tea catechins, gave little effects of cell cycle progression and p21/Waf1 expression. Thus, the present results suggest the difference in the apoptosis-induction mechanism between the two types of tea constituents. PMID- 15806978 TI - Protection of hepatocytes from apoptosis by a novel substance from actinomycetes culture medium. AB - A novel substance, #675, found from an Streptomyces sp. SM675 culture medium, dose-dependently stimulates the proliferation of human functional liver cell 4 (FLC4). When FLC4 cells were incubated under conditions without fetal bovine serum (FBS), typical features of apoptotic cell death such as shrinkage and nuclear condensation appeared; high molecular weight (HMW) DNA fragments were found; and caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were cleaved. When FLC4 cells were incubated with #675 and without FBS, the cells grew healthy, no HMW DNA fragments were found, and caspase-3 and PARP cleavage weakened, suggesting that #675 protects FLC4 cells from apoptosis induced by FBS deprivation. The quantitative reverse-transcribed polymerase chain reaction did not show differences in PARP or Bcl-2 mRNA expression in FLC4 cells incubated with or without #675, indicating other genes may be involved in this anti apoptosis effect. These results show that #675 enhances FLC4 proliferation via an apoptosis-inhibition pathway, implying potential pharmacological and clinical applications. PMID- 15806979 TI - The effect of magnetic stimulation on unloaded soleus muscle of rat: changes in myosin heavy chain mRNA isoforms. AB - This study assessed the potential application and the effectiveness of functional magnetic stimulation (FMS) for preventing skeletal muscle atrophy in adult rats. FMS using magnetic stimulator was performed to rat soleus muscle by placing a round magnetic coil on the back of 3rd-5th lumbar vertebral level at 20 Hz frequency for 60 min/day up to 10 days. A reverse transcriptase-polymerase chain reaction was applied to evaluate relative amounts of mRNAs specific to four myosin heavy chain (MHC) isoforms [MHCIbeta, MHCIIa, MHCIIb, and MHCIId(x)] in rat soleus muscle during contractile activity by magnetic stimulation. Ten-day unloading by hindlimb suspension induced a drastic decrease of MHCIbeta and MHCIIa mRNA expressions, while MHCIIb and MHCIId(x) mRNA was not decreased. The magnetic stimulation resuscitated the down-regulation of the mRNA levels of MHCIbeta and MHCIIa. These results suggest that magnetic stimulation on acute atrophied muscles is useful for preventing the muscle atrophy. PMID- 15806980 TI - Normalization of high-flow or removal of flow cannot stop high-flow induced endothelial proliferation. AB - Endothelial cells (ECs) are activated in response to high-flow. Our previous studies using arteriovenous fistula (AVF) model have demonstrated that high-flow in blood vessels induces an early and rapid proliferation of ECs before arterial dilatation. Here, we investigated the proliferation of ECs, which had once been stimulated by high-flow loading, in a situation without the influence of high flow. First, we induced high-flow in the rabbit common carotid artery by using AVF. Then, we removed the influence of high-flow by normalization of high-flow with the closure of AVF or by removal of flow itself with tissue isolation and organ culture or with cell culture of ECs, at the timing considered that ECs began to proliferate. Kinetics of ECs was investigated by a laser scanning confocal microscopy, phase-contrast microscopy and light microscopy using bromodeoxyuridine labeling method. We found that ECs, which had once been stimulated by high-flow, transiently proliferated even after normalization of high-flow or removal of flow. We assume that proliferation of ECs is promised when these cells start to proliferate after high-flow loading. PMID- 15806981 TI - Loxoprofen inhibits facilitated micturition reflex induced by acetic acid urinary bladder infusion of the rats. AB - Prostaglandins (PGs) are well known as one of the chemical mediators of inflammation. Nonsteroidal anti-inflammatory drugs (NSAIDs), PG synthesis inhibitors, are used for anti-nociception and/or anti-inflammation. We examine the effect of loxoprofen, an NSAID, on micturiton in acetic acid-induced bladder inflammation of the rats. In cystometrogram study with saline infusion into the urinary bladder, loxoprofen did not alter the interval of bladder contraction (IC, 107% of the control). IC was shortened by acetic acid infusion (65% of the control) and loxoprofen prolonged the IC (162% of acetic acid infused period). This prolonged IC was approximately same as the control. Loxoprofen did not alter the threshold pressure and the maximal voiding pressure. These data suggest that PGE2 might not play a part of normal micturition and may play a part of the micturition reflex during acetic acid infusion. That is, loxoprofen might be useful for pathological hyperreflex of the micturition. PMID- 15806982 TI - Augmentation of Helicobacter pylori urease activity by its specific IgG antibody: implications for bacterial colonization enhancement. AB - Gastric colonization of Helicobacter pylori (H. pylori) occurs in a very early age via infected mothers having H. pylori-specific IgG antibodies that would be transplacentally transferred to infants. In addition, H. pylori urease-specific IgG was associated with chronic gastric atrophy and post-immunization gastritis is usually correlated with a strong local IgG response. These findings indicate that H. pylori-specific IgG antibodies, in particular its urease-specific IgG, may induce unfavorable influence on host resistance against H. pylori. Here, we show that we have found a unique H. pylori urease-specific IgG monoclonal antibody (MAb), termed S3, recognizing the conformational structure of the small subunit Ure-A, which enhanced the urease enzymatic activity. Such enhancement of the H. pylori urease activity induced by 1 microg of S3 was almost completely cancelled by simultaneously added the same amount of L2 MAb, which has a strong and specific inhibitory activity against H. pylori urease and recognizes a liner epitope of 8-mer peptide (F8: SIKEDVQF) within its large subunit Ure-B (Infect. Immun. 69: 6597, 2001). Intravenous pre-administration of purified S3 into BALB/c mice showed significant augmentation for gastric colonization with the susceptible strain Sydney Strain-1 (SS-1). To our knowledge, this is the first demonstration that a H. pylori urease-specific IgG MAb induced an augmentation of their gastric colonization in vivo. PMID- 15806983 TI - Death with indignity. PMID- 15806984 TI - Prescription for crime. PMID- 15806985 TI - When bad bugs go good. PMID- 15806986 TI - Hall of shame. PMID- 15806987 TI - Vitamin E-gads. PMID- 15806989 TI - Spectral subtraction-based speech enhancement for cochlear implant patients in background noise. PMID- 15806990 TI - A beamforming video recorder for integrated observations of dolphin behavior and vocalizations. PMID- 15807005 TI - Using listening difficulty ratings of conditions for speech communication in rooms. AB - The use of listening difficulty ratings of speech communication in rooms is explored because, in common situations, word recognition scores do not discriminate well among conditions that are near to acceptable. In particular, the benefits of early reflections of speech sounds on listening difficulty were investigated and compared to the known benefits to word intelligibility scores. Listening tests were used to assess word intelligibility and perceived listening difficulty of speech in simulated sound fields. The experiments were conducted in three types of sound fields with constant levels of ambient noise: only direct sound, direct sound with early reflections, and direct sound with early reflections and reverberation. The results demonstrate that (1) listening difficulty can better discriminate among these conditions than can word recognition scores; (2) added early reflections increase the effective signal-to noise ratio equivalent to the added energy in the conditions without reverberation; (3) the benefit of early reflections on difficulty scores is greater than expected from the simple increase in early arriving speech energy with reverberation; (4) word intelligibility tests are most appropriate for conditions with signal-to-noise (S/N) ratios less than 0 dBA, and where S/N is between 0 and 15-dBA S/N, listening difficulty is a more appropriate evaluation tool. PMID- 15807010 TI - Encoding of vowel-like sounds in the auditory nerve: model predictions of discrimination performance. AB - The sensitivity of listeners to changes in the center frequency of vowel-like harmonic complexes as a function of the center frequency of the complex cannot be explained by changes in the level of the stimulus [Lyzenga and Horst, J. Acoust. Soc. Am. 98, 1943-1955 (1995)]. Rather, a complex pattern of sensitivity is seen; for a spectrum with a triangular envelope, the greatest sensitivity occurs when the center frequency falls between harmonics, whereas for a spectrum with a trapezoidal envelope, greatest sensitivity occurs when the center frequency is aligned with a harmonic. In this study, the thresholds of a population model of auditory-nerve (AN) fibers were quantitatively compared to these trends in psychophysical thresholds. Single-fiber and population model responses were evaluated in terms of both average discharge rate and the combination of rate and timing information. Results indicate that phase-locked responses of AN fibers encode phase transitions associated with minima in these amplitude-modulated stimuli. The temporal response properties of a single AN fiber, tuned to a frequency slightly above the center frequency of the harmonic complex, were able to explain the trends in thresholds for both triangular- and trapezoidal-shaped spectra. PMID- 15807011 TI - Cochlear gain control. AB - The nonlinear auditory phenomena of compression, suppression, and distortion are known to have a cochlear-mechanical origin. An instantaneous nonlinear transfer function is often assumed to underlie these phenomena, but there are experimental indications that auditory nonlinearity is sluggish rather than instantaneous. This study analyzes the consequences of such sluggishness, using automatic gain control (AGC) as a model noninstantaneous nonlinearity. The distinctive characteristic of AGC, its delayed action, is shown to produce a number of observable and measurable effects that distinguish AGC from instantaneous nonlinearities. A major class of such AGC-specific effects concerns the phase of aural distortion products. For example, the phase of the cancellation tone in the classical psychoacoustic cancellation paradigm is linearly related to the frequency spacing of the primary tones in an AGC, as opposed to the square-law relationship produced by an instantaneous nonlinearity. These and other predictions are confronted with experimental data from the literature. The impact of putative AGC-related delays on the interpretation of distortion product otoacoustic emissions (DPOAEs) is discussed. Detailed suggestions are made for experiments specifically aimed at determining whether cochlear nonlinearity is instantaneous or delayed. PMID- 15807012 TI - On the large-scale spectral structure of otoacoustic emissions. AB - Transient evoked and distortion product otoacoustic emission data, showing a characteristic slowly oscillating spectral shape, are presented. Such peculiar behavior had also been observed in earlier studies, and deserves some theoretical explanation. A simple model of the cochlear reflectivity, based on the analogy between the cochlear transmission line equations and the Schrodinger wave equation for the motion of an elementary particle above a one-dimensional potential well, is presented. Wave mechanics predicts indeed reflection from a negative potential well, which is quasiperiodically dependent on the width and depth of the well, i.e., on the quality factor of the cochlear resonance. The model, whose quantitative predictions are dependent on the rather uncertain level and slope of the cochlear tuning curve, proves capable of explaining, at least qualitatively, the observed experimental behavior. PMID- 15807013 TI - Investigation of potential effects of cellular phones on human auditory function by means of distortion product otoacoustic emissions. AB - Outer hair cells (OHC) are thought to act like piezoelectric transducers that amplify low sounds and hence enable the ear's exquisite sensitivity. Distortion product otoacoustic emissions (DPOAE) reflect OHC function. The present study investigated potential effects of electromagnetic fields (EMF) of GSM (Global System for Mobile Communication) cellular phones on OHCs by means of DPOAEs. DPOAE measurements were performed during exposure, i.e., between consecutive GSM signal pulses, and during sham exposure (no EMF) in 28 normally hearing subjects at tone frequencies around 4 kHz. For a reliable DPOAE measurement, a 900-MHz GSM like signal was used where transmission pause was increased from 4.034 ms (GSM standard) to 24.204 ms. Peak transmitter power was set to 20 W, corresponding to a specific absorption rate (SAR) of 0.1 W/kg. No significant change in the DPOAE level in response to the EMF exposure was found. However, when undesired side effects on DPOAEs were compensated, in some subjects an extremely small EMF exposure-correlated change in the DPOAE level (< 1 dB) was observed. In view of the very large dynamic range of hearing in humans (120 dB), it is suggested that this observation is physiologically irrelevant. PMID- 15807014 TI - Distortion-product otoacoustic emission measured with continuously varying stimulus level. AB - Distortion-product otoacoustic emissions (DPOAE) are measured by stimulating the ear with two simultaneous tones. A novel method for measuring DPOAEs has been developed in which the tone levels vary continuously instead of in discrete steps. Varying the tone levels continuously may offer advantages for characterizing DPOAE level as a function of stimulus level. For equivalent primary levels, DPOAE levels measured with the continuous-level method were the same as levels obtained with the discrete-level method, thus validating the new method. Continuous-level measurements were used to determine the optimal L1 for each L2 in individual subjects (N= 20) at f2 = 1, 2, 4, and 8 kHz by using a Lissajous path that covered a wide range of stimulus levels. The optimal L1 (defined as the L1 that resulted in the largest DPOAE for each L2) varied across subjects and across frequency. The optimal difference between L1 and L2 decreased with increasing L2 at all frequencies, and increased with frequency when L2 was low. When the optimal L1 was determined individually for each ear, the DPOAE levels were larger and less variable than those obtained using the equation for L1 suggested by Kummer et al. [J. Acoust. Soc. Am. 103, 3431-3444 (1998)]. PMID- 15807015 TI - The Allen-Fahey experiment extended. AB - An ingenious experiment has been performed by Allen and Fahey [J. Acoust. Soc. Am. 92, 178-188 (1992)], in which they attempted to estimate the gain of the cochlear amplifier by comparing responses to the 2 f1-f2 distortion product (DP) in the outer ear canal (otoacoustic emissions) and from an auditory-nerve fiber. Results were essentially negative: no evidence of cochlear amplification was found in that experiment. A variation of that experiment is reported here, where DP responses in the outer ear canal are compared with mechanical responses of the basilar membrane. This variation does not suffer from the major limitation in the original experiment in the choice of possible frequency ratios. Results confirm and extend those of Allen and Fahey entirely. Apparently, the gain of the cochlear amplifier cannot be measured in this way. It is argued that the retrograde wave going to the stapes is most likely reduced in magnitude by wave interference when the two primary frequencies approach each other. Such a reduction does not take place in the forward-going wave to the location tuned to the DP frequency. This explanation is illustrated on the basis of results of earlier experiments on the movements of the basilar membrane. PMID- 15807016 TI - Recording depth and signal competition in heterodyne interferometry. AB - A common way to measure submicroscopic motion of the organ of Corti is heterodyne interferometry. The depth over which vibration can be accurately measured with heterodyne interferometry is determined by both the optics, which controls to what extent light from nonfocal planes reaches the photodetectors, and demodulation electronics, which determines to what extent signal generated by out of-focal-plane light influences the measurements. The influence of a second reflecting surface is investigated theoretically and experimentally. By reviewing the theory of FM demodulation and showing tests with a Revox FM demodulator, it is demonstrated that the influence of a secondary signal on a measurement depends on the modulation index. Both high- and low-modulation index signals are encountered in heterodyne interferometry of the cochlea. Using a He-Ne-like diode laser (lambda = 638 nm), the border between low- and high-modulation signals is at a displacement of about 25-100 nm. Confocal interferometry reduces the magnitude of out-of-focus signals, and therefore their effect on vibration measurement. The response of the confocal system to reflected signals from two surfaces separated by distances encountered within the cochlear partition is shown. The results underscore the benefit of steep optical sectioning for intracochlear measurements. PMID- 15807017 TI - Integration and segregation in auditory scene analysis. AB - Assessment of the neural correlates of auditory scene analysis, using an index of sound change detection that does not require the listener to attend to the sounds [a component of event-related brain potentials called the mismatch negativity (MMN)], has previously demonstrated that segregation processes can occur without attention focused on the sounds and that within-stream contextual factors influence how sound elements are integrated and represented in auditory memory. The current study investigated the relationship between the segregation and integration processes when they were called upon to function together. The pattern of MMN results showed that the integration of sound elements within a sound stream occurred after the segregation of sounds into independent streams and, further, that the individual streams were subject to contextual effects. These results are consistent with a view of auditory processing that suggests that the auditory scene is rapidly organized into distinct streams and the integration of sequential elements to perceptual units takes place on the already formed streams. This would allow for the flexibility required to identify changing within-stream sound patterns, needed to appreciate music or comprehend speech. PMID- 15807018 TI - Combined representations for frequency and duration in detection templates for expected signals. AB - When trying to detect a tonal signal in a continuous broadband noise, listeners attend selectively to both the frequency and the duration of the expected signal. However, it is not known whether they monitor separate or combined representations of these two attributes. To investigate this question, a probe signal method was used to measure the detectability of signals of expected and unexpected durations at two expected frequencies. The four listeners expected only one of two signals to be presented at random: a brief tone at one frequency or a long tone at another frequency. For each signal frequency, the detectability of the signals of unexpected duration decreased to near chance as the difference between the expected and unexpected duration, at that frequency, increased. The frequency specificity of this duration tuning indicates that both the frequency and the duration of an expected stimulus are represented in a single template. PMID- 15807019 TI - Detection-theoretic analysis of same-different judgments for the amplitude discrimination of acoustic sinusoids. AB - Three methods were applied to determine which decision model best accounted for same-different judgments about the amplitude of acoustic sinusoids. The methods were (1) analysis of the shape of the receiver operating characteristic; (2) analysis of an observer's decision space; and (3) a correlation method based on the conditional-on-single-stimulus procedure. In one experiment, observers rated their confidence that a pair of 1-kHz sinusoids was drawn from the same or from different sets. The two sets had identical Gaussian distributions of decibel amplitudes, but their mean amplitudes differed by 3 dB. The differencing model, in which observers base their decision on the absolute difference between the two observations, accounted for the data better than the independent-observation model, in which observers compute independent likelihood ratios for each observation. A second experiment added to every trial an interval that contained a 1-kHz sinusoid whose amplitude equaled the grand mean of both stimulus sets. Despite this additional information, which is needed to adopt the independent observation model, the differencing model again better accounted for the data. PMID- 15807020 TI - Effect of frequency-modulation coherence for inharmonic stimuli: frequency modulation phase discrimination and identification of artificial double vowels. AB - The ability to compare patterns of frequency modulation (FM) in separate frequency regions was explored. In experiment 1, listeners had to distinguish whether the FM applied to two nonharmonically related sinusoidal carriers was in phase or out of phase. The FM rate was the same for each carrier. The starting phase of the modulation was randomized for each stimulus in a three alternative, forced-choice (3AFC) trial. Subjects were sensitive to relative FM phase for modulation rates of 2 and 4 Hz, but not for higher rates. In experiment 2, vowel identification was compared for artificial single and double vowels. The vowels were constructed from complex tones with components spaced at 2-ERB(N) (equivalent rectangular bandwidth) intervals, by increasing the levels of three components by 15 dB, to create three "formants." In the double vowels, the components of the two vowels were interleaved, to give 1-ERB(N) spacing. The three "formant" components were frequency modulated at 2, 4, or 8 Hz, with either the same or different rates for the two vowels. The identification of double vowels was not improved by a difference in FM rate across vowels, suggesting that differences in FM rate do not support perceptual segregation of inharmonic stimuli. PMID- 15807021 TI - Dominance region for pitch: effects of duration and dichotic presentation. AB - The dominance region (DR) for pitch was determined for 16- and 200-ms complex tones containing the first seven harmonics of a fundamental frequency (F0) of 250 Hz. A tone was presented with one of the harmonics mistuned upwards or downwards by 3%, followed 500 ms later by a perfectly harmonic tone of the same duration. Listeners adjusted the F0 of the harmonic tone so that its pitch matched that of the mistuned complex. In experiment 1, stimuli were presented monaurally. The DR was significantly higher in harmonic number for the short than for the long duration. The overall sum of the pitch shifts produced by all harmonics was significantly larger for the short than for the long duration, presumably due to stronger perceptual fusion for the former. In experiment 2, the mistuned harmonic was presented only contralaterally to the remainder of the complex. A similar shift in the DR with duration was observed, although the pitch shifts were smaller than for monaural presentation. There was no significant effect of duration on the overall pitch shifts. The results are discussed in terms of pattern recognition and autocorrelation models of pitch perception, and a role of attention in pitch matching is suggested. PMID- 15807022 TI - Frequency dependency of the relationship between perceived auditory source width and the interaural cross-correlation coefficient for time-invariant stimuli. AB - Previous research has indicated that the relationship between the interaural cross-correlation coefficient (IACC) of a narrow-band sound and its perceived auditory source width is dependent on its frequency. However, this dependency has not been investigated in sufficient detail for researchers to be able to properly model it in order to produce a perceptually relevant IACC-based model of auditory source width. A series of experiments has therefore been conducted to investigate this frequency dependency in a controlled manner, and to derive an appropriate model. Three main factors were discovered in the course of these experiments. First, the nature of the frequency dependency of the perceived auditory source width of stimuli with an IACC of 1 was determined, and an appropriate mathematical model was derived. Second, the loss of perceived temporal detail at high frequencies, caused by the breakdown of phase locking in the ear, was found to be relevant, and the model was modified accordingly using rectification and a low-pass filter. Finally, it was found that there was a further frequency dependency at low frequencies, and a method for modeling this was derived. The final model was shown to predict the experimental data well. PMID- 15807023 TI - Speech and melody recognition in binaurally combined acoustic and electric hearing. AB - Speech recognition in noise and music perception is especially challenging for current cochlear implant users. The present study utilizes the residual acoustic hearing in the nonimplanted ear in five cochlear implant users to elucidate the role of temporal fine structure at low frequencies in auditory perception and to test the hypothesis that combined acoustic and electric hearing produces better performance than either mode alone. The first experiment measured speech recognition in the presence of competing noise. It was found that, although the residual low-frequency (<1000 Hz) acoustic hearing produced essentially no recognition for speech recognition in noise, it significantly enhanced performance when combined with the electric hearing. The second experiment measured melody recognition in the same group of subjects and found that, contrary to the speech recognition result, the low-frequency acoustic hearing produced significantly better performance than the electric hearing. It is hypothesized that listeners with combined acoustic and electric hearing might use the correlation between the salient pitch in low-frequency acoustic hearing and the weak pitch in the envelope to enhance segregation between signal and noise. The present study suggests the importance and urgency of accurately encoding the fine-structure cue in cochlear implants. PMID- 15807024 TI - Simulations of temporal patterns of oral airflow in men and women using a two mass model of the vocal folds under dynamic control. AB - In this study we use a low-dimensional laryngeal model to reproduce temporal variations in oral airflow produced by speakers in the vicinity of an abduction gesture. It attempts to characterize these temporal patterns in terms of biomechanical parameters such as glottal area, vocal fold stiffness, subglottal pressure, and gender differences in laryngeal dimensions. A two-mass model of the vocal folds coupled to a two-tube approximation of the vocal tract is fitted to oral airflow records measured in men and women during the production of /aha/ utterances, using the subglottal pressure, glottal width, and Q factor as control parameters. The results show that the model is capable of reproducing the airflow records with good approximation. A nonlinear damping characteristics is needed, to reproduce the flow variation at glottal abduction. Devoicing is achieved by the combined action of vocal fold abduction, the decrease of subglottal pressure, and the increase of vocal fold tension. In general, the female larynx has a more restricted region of vocal fold oscillation than the male one. This would explain the more frequent devoicing in glottal abduction-adduction gestures for /h/ in running speech by women, compared to men. PMID- 15807025 TI - Measurements of glottal structure dynamics. AB - Low power, radarlike electromagnetic (EM) wave sensors, operating in a homodyne interferometric mode, are being used to measure tissue motions in the human vocal tract during speech. However, when these and similar sensors are used in front of the laryngeal region during voiced speech, there remains an uncertainty regarding the contributions to the sensor signal from vocal fold movements versus those from pressure induced trachea-wall movements. Several signal-source hypotheses are tested by performing experiments with a subject who had undergone tracheostomy, and who still was able to phonate when her stoma was covered (e.g., with a plastic plate). Laser-doppler motion-measurements of the subject's posterior trachea show small tissue movements, about 15 microns, that do not contribute significantly to signals from presently used EM sensors. However, signals from the anterior wall do contribute. EM sensor and air-pressure measurements, together with 3-D EM wave simulations, show that EM sensors measure movements of the vocal folds very well. The simulations show a surprisingly effective guiding of EM waves across the vocal fold membrane, which, upon glottal opening, are interrupted and reflected. These measurements are important for EM sensor applications to speech signal de-noising, vocoding, speech recognition, and diagnostics. PMID- 15807026 TI - Estimation of sound pressure levels of voiced speech from skin vibration of the neck. AB - How accurately can sound pressure levels (SPLs) of speech be estimated from skin vibration of the neck? Measurements using a small accelerometer were carried out in 27 subjects (10 males and 17 females) who read Rainbow and Marvin Williams passages in soft, comfortable, and loud voice, while skin acceleration levels (SALs) and SPLs were simultaneously registered and analyzed every 30 ms. The results indicate that the mean SPL of voiced speech can be estimated with accuracy better than +/-2.8 dB in 95% of the cases when the subjects are individually calibrated. This makes the accelerometer an interesting sensor for SPL measurement of speech when microphones are problematic to use (e.g., noisy environments or in voice dosimetry). The estimates of equivalent SPL, which is the logarithm of averaged relative energy of voiced speech, were found to be up to 1.5 dB less accurate than the mean SPL. The estimation accuracy for instantaneous SPLs was worse than for the mean and equivalent SPLs (on average +/ 6 and +/-5 dB for males and females, respectively). PMID- 15807027 TI - The relative importance of spectral tilt in monophthongs and diphthongs. AB - Ito et al. [J. Acoust. Soc. Am. 110, 1141-1149 (2001)] demonstrated that listeners can reliably identify vowel stimuli on the basis of relative formant amplitude in the absence of, or in spite of, F2 peak frequency. In the present study, formant frequencies and global spectral tilt are manipulated independently in synthetic steady-state vowels. Listeners' identification of these sounds demonstrate strong perceptual effects for both local (formant frequency) and global (spectral tilt) acoustic characteristics. Subsequent experiments reveal that effects of spectral tilt are attenuated in synthetic stimuli for which formant center frequencies change continuously. When formant peaks are kinematic, perceptual salience of the relative amplitudes of low- and high-frequency formants (as determined by spectral tilt) is mitigated. Because naturally produced English vowels are rarely spectrally static, one may conclude that gross spectral properties may play only a limited role in perception of fluently produced vowel sounds. PMID- 15807028 TI - Interactions between cochlear implant electrode insertion depth and frequency place mapping. AB - While new electrode designs allow deeper insertion and wider coverage in the cochlea, there is still considerable variation in the insertion depth of the electrode array among cochlear implant users. The present study measures speech recognition as a function of insertion depth, varying from a deep insertion of 10 electrodes at 28.8 mm to a shallow insertion of a single electrode at 7.2 mm, in four Med-El Combi 40+ users. Short insertion depths were simulated by inactivating apical electrodes. Speech recognition increased with deeper insertion, reaching an asymptotic level at 21.6 or 26.4 mm depending on the frequency-place map used. Baskent and Shannon [J. Acoust. Soc. Am. 116, 3130-3140 (2004)] showed that speech recognition by implant users was best when the acoustic input frequency was matched onto the cochlear location that normally processes that frequency range, minimizing the spectral distortions in the map. However, if an electrode array is not fully inserted into the cochlea, a matched map will result in the loss of considerable low-frequency information. The results show a strong interaction between the optimal frequency-place mapping and electrode insertion depth. Consistent with previous studies, frequency-place matching produced better speech recognition than compressing the full speech range onto the electrode array for full insertion ranges (20 to 25 mm from the round window). For shallower insertions (16.8 and 19.2 mm) a mild amount of frequency-place compression was better than truncating the frequency range to match the basal cochlear location. These results show that patients with shallow electrode insertions might benefit from a map that assigns a narrower frequency range than patients with full insertions. PMID- 15807029 TI - Glottal open quotient in singing: measurements and correlation with laryngeal mechanisms, vocal intensity, and fundamental frequency. AB - This article presents the results of glottal open-quotient measurements in the case of singing voice production. It explores the relationship between open quotient and laryngeal mechanisms, vocal intensity, and fundamental frequency. The audio and electroglottographic signals of 18 classically trained male and female singers were recorded and analyzed with regard to vocal intensity, fundamental frequency, and open quotient. Fundamental frequency and open quotient are derived from the differentiated electroglottographic signal, using the DECOM (DEgg Correlation-based Open quotient Measurement) method. As male and female phonation may differ in respect to vocal-fold vibratory properties, a distinction is made between two different glottal configurations, which are called laryngeal mechanisms: mechanism 1 (related to chest, modal, and male head register) and mechanism 2 (related to falsetto for male and head register for female). The results show that open quotient depends on the laryngeal mechanisms. It ranges from 0.3 to 0.8 in mechanism 1 and from 0.5 to 0.95 in mechanism 2. The open quotient is strongly related to vocal intensity in mechanism 1 and to fundamental frequency in mechanism 2. PMID- 15807030 TI - Estimation of total attenuation and scatterer size from backscattered ultrasound waveforms. AB - Quantitative ultrasound techniques using backscattered echoes have had limited success in vivo due to the frequency-dependent attenuation along the entire propagation path masking the frequency dependence of the backscatter. Herein, total attenuation and scatterer size are estimated simultaneously by an analysis of the in vivo backscattered power spectrum using two approaches. The simulations used to evaluate the two approaches used frequencies between 4 and 11 MHz with an effective scatterer radius of 25 microm. The first approach was based on approximations of the in vivo backscattered power spectrum (i.e., assumed Gaussian function), wherein attenuation and size were estimated by assuming each was a Gaussian transformation performed on Gaussian power spectra. The approach had poor accuracy due to the backscattered power spectra not being sufficiently modeled by a Gaussian function. The second approach estimated attenuation and size by fitting a modified reference spectrum to the in vivo backscattered power spectrum without any assumptions about the shape of the spectrum. The accuracy of the size estimate was better than 20% for signal-to-noise ratio >6 dB, window lengths greater than 4 mm, and attenuation between 0 and 1 dB/cm-MHz. However, the precision quickly degraded with increasing noise, increasing attenuation, and decreasing window length. PMID- 15807031 TI - Ultrasonic excitation of a bubble near a rigid or deformable sphere: implications for ultrasonically induced hemolysis. AB - A number of independent studies have reported increased ultrasound bioeffects, such as hemolysis and hemorrhage, when ultrasound contrast agents are present. To better understand the role of cavitation in these bioeffects, one- and two dimensional models have been developed to investigate the interactions between ultrasonically excited bubbles and model "cells." First, a simple one-dimensional model based on the Rayleigh-Plesset equation was developed to estimate upper bounds for strain, strain rate, and areal expansion of a simulated red blood cell. Then, two-dimensional boundary element models were developed (with DynaFlow Inc.) to obtain simulations of asymmetric bubble dynamics in the presence of rigid and deformable spheres. The deformable spherical "cell" was modeled using Tait's equation of state for water, with a membrane approximated by surface tension that increases linearly with areal expansion. The presence of a rigid or deformable sphere had little effect on the bubble expansion, but caused an asymmetric collapse and jetting for the conditions considered. Predicted membrane areal expansions were found to be below critical values for hemolysis reported in the literature for the cases considered near the inertial cavitation threshold. PMID- 15807032 TI - Characteristics of the audio sound generated by ultrasound imaging systems. AB - Medical ultrasound scanners use high-energy pulses to probe the human body. The radiation force resulting from the impact of such pulses on an object can vibrate the object, producing a localized high-intensity sound in the audible range. Here, a theoretical model for the audio sound generated by ultrasound scanners is presented. This model describes the temporal and spectral characteristics of the sound. It has been shown that the sound has rich frequency components at the pulse repetition frequency and its harmonics. Experiments have been conducted in a water tank to measure the sound generated by a clinical ultrasound scanner in various operational modes. Results are in general agreement with the theory. It is shown that a typical ultrasound scanner with a typical spatial-peak pulse average intensity value at 2 MHz may generate a localized sound-pressure level close to 100 dB relative to 20 microPa in the audible (< 20 kHz) range under laboratory conditions. These findings suggest that fetuses may become exposed to a high-intensity audio sound during maternal ultrasound examinations. Therefore, contrary to common beliefs, ultrasound may not be considered a passive tool in fetal imaging. PMID- 15807033 TI - Whistles of tucuxi dolphins (Sotalia fluviatilis) in Brazil: comparisons among populations. AB - A quantitative and comparative analysis of the acoustic whistles parameters of tucuxi dolphins (Sotalia fluviatilis) from eight areas in Brazilian waters were conducted. Tucuxi produced mainly upsweep whistles, and despite their similar characteristics to other delphinids, whistles were shorter and less complex in shape. Some signals had fundamental frequency components above 24 kHz, and a broader band system may be useful in further studies concerning tucuxi whistles. The frequency parameters of the whistles had the lowest variation coefficients, whereas the duration and number of inflections had the highest variation coefficients. About 65% of all whistles had common characteristics between study localities, but SF, MinF, MeF, F1/4, F1/2, and F3/4 increased in frequency from southern to northern populations. The magnitude of the whistle characteristics variation was comparatively smaller between adjacent sites areas than between nonadjacent ones. However, for some pairwise comparisons the whistles were more similar between nonadjacent areas than between adjacent ones, indicating that other factors besides genetic and mixing affected tucuxi whistle characteristics. Only a few numbers of whistle variables were significantly different between adjacent areas and the parameter ranges of whistles overlapped, which made it difficult to discriminate whistles between adjacent tucuxi populations. PMID- 15807034 TI - Underwater, low-frequency noise in a coastal sea turtle habitat. AB - Underwater sound was recorded in one of the major coastal foraging areas for juvenile sea turtles in the Peconic Bay Estuary system in Long Island, New York. The recording season of the underwater environment coincided with the sea turtle activity season in an inshore area where there is considerable boating and recreational activity, especially during the summer between Independence Day and Labor Day. Within the range of sea turtle hearing, average noise pressure reached 110 dB during periods of high human activity and diminished proportionally, down to 80 dB, with decreasing human presence. Therefore, during much of the season when sea turtles are actively foraging in New York waters, their coastal habitats are flooded with underwater noise. During the period of highest human activity, average noise pressures within the range of frequencies heard by sea turtles were greater by over two orders of magnitude (26 dB) than during the lowest period of human activity. Sea turtles undoubtedly are exposed to high levels of noise, most of which is anthropogenic. Results suggest that continued exposure to existing high levels of pervasive anthropogenic noise in vital sea turtle habitats and any increase in noise could affect sea turtle behavior and ecology. PMID- 15807035 TI - Three-dimensional beam pattern of regular sperm whale clicks confirms bent-horn hypothesis. AB - The three-dimensional beam pattern of a sperm whale (Physeter macrocephalus) tagged in the Ligurian Sea was derived using data on regular clicks from the tag and from hydrophones towed behind a ship circling the tagged whale. The tag defined the orientation of the whale, while sightings and beamformer data were used to locate the whale with respect to the ship. The existence of a narrow, forward-directed P1 beam with source levels exceeding 210 dBpeak re: 1 microPa at 1 m is confirmed. A modeled forward-beam pattern, that matches clicks >20 degrees off-axis, predicts a directivity index of 26.7 dB and source levels of up to 229 dBpeak re: 1 microPa at 1 m. A broader backward-directed beam is produced by the P0 pulse with source levels near 200 dBpeak re: 1 microPa at 1 m and a directivity index of 7.4 dB. A low-frequency component with source levels near 190 dBpeak re: 1 microPa at 1 m is generated at the onset of the P0 pulse by air resonance. The results support the bent-horn model of sound production in sperm whales. While the sperm whale nose appears primarily adapted to produce an intense forward-directed sonar signal, less-directional click components convey information to conspecifics, and give rise to echoes from the seafloor and the surface, which may be useful for orientation during dives. PMID- 15807036 TI - Indication of a Lombard vocal response in the St. Lawrence River Beluga. AB - Noise pollution is recognized as a potential danger to marine mammals in general, and to the St. Lawrence beluga in particular. One method of determining the impacts of noise on an animal's communication is to observe a natural and repeatable response of the vocal system to variations in noise level. This is accomplished by observing intensity changes in animal vocalizations in response to environmental noise. One such response observed in humans, songbirds, and some primates is the Lombard vocal response. This response represents a vocal system reaction manifested by changes in vocalization level in direct response to changes in the noise field. In this research, a population of belugas in the St. Lawrence River Estuary was tested to determine whether a Lombard response existed by using hidden Markhov-classified vocalizations as targets for acoustical analyses. Correlation and regression analyses of signals and noise indicated that the phenomenon does exist. Further, results of human subjects experiments [Egan, J. J. (1966), Ph.D. dissertation; Scheifele, P. M. (2003), Ph.D. dissertation], along with previously reported data from other animal species, are similar to those exhibited by the belugas. Overall, findings suggest that typical noise levels in the St. Lawrence River Estuary have a detectable effect on the communication of the beluga. PMID- 15807037 TI - Bird's-eye view. Ultraviolet vision is commonplace in the animal world. A very few people, it turns out, have this ability too. PMID- 15807038 TI - Strategic intensity: a conversation with world chess champion Garry Kasparov. AB - It's hard to find a better exemplar for competition than chess. The image of two brilliant minds locked in a battle of skill and will-in which chance plays little or no apparent role-is compelling. Even people who have scant knowledge of the game instinctively recognize that chess is unusual in terms of its intellectual complexity and the strategic demands it places on players. Can strategists learn anything from chess players about what it takes to win? To find out, H BR senior editor Diane L. Coutu talked with Garry Kasparov, the world's number one player since 1984. Kasparov believes that success in both chess and business is very much a question of psychological advantage; the complexity of the game demands that players rely heavily on their instincts and on gamesmanship. In this wide ranging interview, Kasparov explores the power of chess as a model for business competition; the balance that chess players strike between intuition and analysis; the significance of his loss to IBM's chess-playing computer, Deep Blue; and how his legendary rivalry with Anatoly Karpov, Kasparov's predecessor as World Chess Champion, affected his own success. Kasparov also shares his solution to what he calls the champion's dilemma, a question for all world masters, whether they are in business, sports, or chess: Where does a virtuoso go after he has accomplished everything he's ever wanted to, even beyond his wildest imagination? If you are lucky, says Kasparov, your enemies will push you to be passionate about staying at the top. PMID- 15807039 TI - How strategists really think. Tapping the power of analogy. AB - Leaders tend to be so immersed in the specifics of strategy that they rarely stop to think how much of their reasoning is done by analogy. As a result, they miss useful insights that psychologists and other scientists have generated about analogies' pitfalls. Managers who pay attention to their own analogical thinking will make better strategic decisions and fewer mistakes. Charles Lazarus was inspired by the supermarket when he founded Toys R Us; Intel promoted its low-end chips to avoid becoming like U.S. Steel; and Circuit City created CarMax because it saw the used-car market as analogous to the consumer-electronics market. Each example displays the core elements of analogical reasoning: a novel problem or a new opportunity, a specific prior context that managers deem to be similar in its essentials, and a solution that managers can transfer from its original setting to the new one. Analogical reasoning is a powerful tool for sparking breakthrough ideas. But dangers arise when analogies are built on surface similarities (headlong diversification based on loose analogies played a role in Enron's collapse, for instance). Psychologists have discovered that it's all too easy to overlook the superficiality of analogies. The situation is further complicated by people's tendency to hang on to beliefs even after contrary evidence comes along (a phenomenon known as anchoring) and their tendency to seek only the data that confirm their beliefs (an effect known as the confirmation bias). Four straightforward steps can improve a management team's odds of using an analogy well: Recognize the analogy and identify its purpose; thoroughly understand its source; determine whether the resemblance is more than superficial; and decide whether the original strategy, properly translated, will work in the target industry. PMID- 15807040 TI - 7 transformations of leadership. AB - Most developmental psychologists agree that what differentiates one leader from another is not so much philosophy of leadership, personality, or style of management. Rather, it's internal "action logic"--how a leader interprets the surroundings and reacts when his or her power or safety is challenged. Relatively few leaders, however, try to understand their action logic, and fewer still have explored the possibility of changing it. They should, because leaders who undertake this voyage of personal understanding and development can transform not only their own capabilities but also those of their companies. The authors draw on 25 years of consulting experience and collaboration with psychologist Susanne Cook-Greuter to present a typology of leadership based on the way managers personally make sense of the world around them. Rooke and Torbert classify leaders into seven distinct actionlogic categories: Opportunists, Diplomats, Experts, Achievers, Individualists, Strategists, and Alchemists-the first three associated with below-average performance, the latter four with medium to high performance. These leadership styles are not fixed, the authors say, and executives who are willing to work at developing themselves and becoming more self-aware can almost certainly move toward one of the more effective action logics. A Diplomat, for instance, can succeed through hard work and self reflection at transforming himself into a Strategist. Few people may become Alchemists, but many will have the desire and potential to become Individualists and Strategists. Corporations that help their executives and leadership teams to examine their action logics can reap rich rewards. PMID- 15807041 TI - Countering the biggest risk of all. AB - Corporate treasurers and chief financial officers have become adept at quantifying and managing a wide variety of risks: financial (for example, currency fluctuations), hazard (chemical spills), and operational (computer system failures). To defend themselves, they use tried-and-true tools such as hedging, insurance, and backup systems. Some companies have even adopted the concept of enterprise risk management, integrating available risk management techniques in a comprehensive, organization-wide approach. But most managers have not addressed in a systematic way the greatest threat of all--strategic risks, the array of external events and trends that can devastate a company's growth trajectory and shareholder value. Strategic risks go beyond such familiar challenges as the possible failure of an acquisition or a product launch. A new technology may overtake your product. Gradual shifts in the market may slowly erode one of your brands beyond the point of viability. Or rapidly shifting customer priorities may suddenly change your industry. The key to surviving these strategic risks, the authors say, is knowing how to assess and respond to them. In this article, they lay out a method for identifying and responding to strategic threats. They categorize the risks into seven major classes (industry, technology, brand, competitor, customer, project, and stagnation) and describe a particularly dangerous example within each category. The authors also offer countermeasures to take against these risks and describe how individual companies (American Express, Coach, and Air Liquide, among them) have deployed them to neutralize a threat and, in many cases, capitalize on it. Besides limiting the downside of risk, strategic-risk management forces executives to think more systematically about the future, thus helping them identify opportunities for growth. PMID- 15807042 TI - The quest for customer focus. AB - Companies have poured enormous amounts of money into customer relationship management, but in many cases the investment hasn't really paid off. That's because getting closer to customers isn't about building an information technology system. It's a learning journey-one that unfolds over four stages, requiring people and business units to coordinate in progressively more sophisticated ways. The journey begins with the creation of a companywide repository containing each interaction a customer has with the company, organized not by product, purchase, or location, but by customer. Communal coordination is what's called for at this stage, as each group contributes its information to the data pool separately from the others and then taps into it as needed. In the second stage, one-way serial coordination from centralized IT through analytical units and out to the operating units allows companies to go beyond just assembling data to drawing inferences. In stage three, companies shift their focus from past relationships to future behavior. Through symbiotic coordination, information flows back and forth between central analytic units and various organizational units like marketing, sales, and operations, as together they seek answers to questions like "How can we prevent customers from switching to a competitor?" and "Who would be most likely to buy a new product in the future"? In stage four, firms begin to move past discrete, formal initiatives and, through integral coordination, bring an increasingly sophisticated understanding oftheir customers to bear in all day-to-day operations. Skipping stages denies organizations the sure foundation they need to build a lasting customer-focused mind-set. Those that recognize this will invest their customer relationship dollars much more wisely-and will see their customer-focusing efforts pay offon the bottom line. PMID- 15807043 TI - The relative value of growth. AB - Most executives would say that adding a point of growth and gaining a point of operating-profit margin contribute about equally to shareholder value. Margin improvements hit the bottom line immediately, while growth compounds value over time. But the reality is that the two are rarely equivalent. Growth often is far more valuable than managers think. For some companies, convincing the market that they can grow by just one additional percentage point can be worth six, seven, or even ten points of margin improvement. This article presents a new strategic metric, called the relative value of growth (RVG), which gives managers a clear picture of how growth projects and margin improvement initiatives affect shareholder value. Using basic balance sheet and income sheet data, managers can determine their companies' RVGs, as well as those of their competitors. Calculating RVGs gives managers insights into which corporate strategies are working to deliver value and whether their companies are pulling the most powerful value-creation levers. The author examines a number of well-known companies and explains what their RVG numbers say about their strategies. He reviews the unspoken assumption that growth and profits are incompatible over the long term and shows that a fair number of companies are effective at delivering both. Finally, he explains how managers can use the RVG framework to help them define strategies that balance growth and profitability at both the corporate and business unit levels. PMID- 15807044 TI - Selection bias and the perils of benchmarking. AB - To find the secrets of business success, what could be more natural than studying successful businesses? In fact, nothing could be more dangerous, warns this Stanford professor. Generalizing from the examples of successful companies is like generalizing about New England weather from data taken only in the summer. That's essentially what businesspeople do when they learn from good examples and what consultants, authors, and researchers do when they study only existing companies or--worse yet--only high-performing companies. They reach conclusions from unrepresentative data samples, falling into the classic statistical trap of selection bias. Drawing on a wealth of case studies, for instance, one researcher concluded that great leaders share two key traits: They persist, often despite initial failures, and they are able to persuade others to join them. But those traits are also the hallmarks of spectacularly unsuccessful entrepreneurs, who must persist in the face of failure to incur large losses and must be able to persuade others to pour their money down the drain. To discover what makes a business successful, then, managers should look at both successes and failures. Otherwise, they will overvalue risky business practices, seeing only those companies that won big and not the ones that lost dismally. They will not be able to tell if their current good fortune stems from smart business practices or if they are actually coasting on past accomplishments or good luck. Fortunately, economists have developed relatively simple tools that can correct for selection bias even when data about failed companies are hard to come by. Success may be inspirational, but managers are more likely to find the secrets of high performance if they give the stories of their competitors'failures as full a hearing as they do the stories of dazzling successes. PMID- 15807045 TI - The half-truth of first-mover advantage. AB - Many executives take for granted that the first company in a new product category gets an unbeatable head start and reaps long-lasting benefits. But that doesn't always happen. The authors of this article discovered that much depends on the pace at which the category's technology is changing and the speed at which the market is evolving. By analyzing these two factors, companies can improve their odds of succeeding as first movers with the resources they possess. Gradual evolution in both the technology and the market provides a first mover with the best conditions for creating a dominant position that is long lasting (Hoover in the vacuum cleaner industry is a good example). In such calm waters, a company can defend its advantages even without exceptional skills or extensive financial resources. When the market is changing rapidly and the product isn't, a first entrant with extensive resources can obtain a long-lasting advantage (as Sony did with its Walkman personal stereo); a company with only limited resources probably must settle for a short-term benefit. When the market is static but the product is changing constantly, first-mover advantages of either kind--durable or short lived--are unlikely. Only companies with very deep pockets can survive (think of Sony and the digital cameras it pioneered). Rapid churn in both the technology and the market creates the worst conditions. But if companies have an acute sense of when to exit-as Netscape demonstrated when it agreed to be acquired by AOL-a worthwhile short-term gain is possible. Before venturing into a newly forming market, you need to analyze the environment, assess your resources, then determine which type offirst-mover advantage is most achievable. Once you've gone into the water, you have no choice but to swim. PMID- 15807046 TI - Sensitivity of distortion product otoacoustic emissions in noise-exposed chinchillas. AB - The present study investigates the effect of small amounts of outer hair cell (OHC) loss on distortion product otoacoustic emission (DPOAE) levels and evoked potential permanent threshold shifts (PTS) in a population of 12 noise-exposed chinchillas. The group mean DPOAE level, which decreased by up to approximately 15 dB in the presence of less than 8 dB PTS and 15% OHC loss, indicates that DPOAEs can detect an underlying cochlear pathology (i,e., OHC damage/loss) despite the presence of normal to near normal thresholds. The sensitivity of DPOAEs in detecting OHC loss makes this test measure suited for diagnosing sensorineural hearing impairment, particularly when abnormal auditory symptoms (i.e., speech discrimination problems) are associated with a normal audiogram in the clinical setting and as part of a hearing conservation program. PMID- 15807047 TI - The instructional effectiveness of a web-based audiometry simulator. AB - With distance learning becoming more of a reality than a novelty in many undergraduate and graduate training programs, web-based clinical simulations can be identified as an instructional option in distance education that has both a sound pedagogical foundation and clinical relevance. The purpose of this article is to report on the instructional effectiveness of a web-based pure-tone audiometry simulator by undergraduate and graduate students in speech-language pathology. Graduate and undergraduate majors in communication sciences and disorders practiced giving basic hearing tests on either a virtual web-based audiometer or a portable audiometer. Competencies in basic testing skills were evaluated for each group. Results of our analyses of the data indicate that both undergraduate and graduate students learned basic audiometric testing skills using the virtual audiometer. These skills were generalized to basic audiometric testing skills required of a speech language pathologist using a portable audiometer. PMID- 15807048 TI - Estimates of loudness, loudness discomfort, and the auditory dynamic range: normative estimates, comparison of procedures, and test-retest reliability. AB - The purpose of this series of experiments was to establish normative reference values for absolute and relative judgements of loudness discomfort and for the auditory dynamic range (DR), and to evaluate intersubject variability and intra subject test-retest reliability for the respective measures of loudness discomfort. To establish the normal auditory DR, audiometric thresholds and loudness discomfort levels (LDLs) were measured from a group of 59 normal-hearing adults without sound tolerance problems. The resulting estimates of the LDL and DR were on the order of 100 dB HL and 95 dB, respectively. A subset (n = 18) of this larger group participated in further studies in which loudness growth functions and the upper limit of the auditory DR were measured by categorical scaling judgments. The findings revealed no significant differences between the test methods for absolute (LDL) and relative (categorical scaling) judgements of loudness discomfort, intersubject variability, or intrasubject test-retest reliability, and suggest that the simple LDL estimate of loudness discomfort is an efficient and valid clinical measure for characterizing the "threshold of discomfort." PMID- 15807049 TI - The effects of expansion on the objective and subjective performance of hearing instrument users. AB - The present study investigated the effects of expansion on the objective and subjective performance of 20 hearing instrument users fitted binaurally with digital ITE products. Objective performance was evaluated in quiet using the Connected Speech Test and in noise using the Hearing in Noise Test. Subjective performance was evaluated in two ways: (a) by having each participant rate their satisfaction regarding the amount of noise reduction they perceived in each expansion condition on a daily basis and (b) by having each participant indicate which expansion condition they preferred following the completion of a two-week trial. Results indicated that expansion significantly reduced low-level speech perception performance; however, satisfaction and preference ratings significantly increased when using expansion. The effect of degree of hearing loss, expansion kneepoint, and expansion ratio on the effectiveness of expansion for a given listener was discussed. PMID- 15807050 TI - Development of low-frequency tone burst versus the click auditory brainstem response. AB - Often ABR threshold testing employs clicks to assess high-frequency hearing, and low-frequency tone bursts to assess low-frequency sensitivity. While a maturation effect has been shown for click stimuli, similar data are lacking for low frequency toneburst stimuli. Thus, 305 infants ranging in conceptional age (CA) from 33 weeks to 74 weeks were tested. Absolute latencies were measured for wave V at 55, 35, and 25 dB nHL in response to a click and for wave V500 in response to a 500 Hz tone burst. Major wave latency in response to 500 Hz tone bursts decreases with age and do not stabilize by 70 weeks CA. Likewise, waves III and V latencies in response to clicks decrease with age, as has been reported by others, and do not stabilize by 70 weeks CA. Wave I latency produced by clicks did not decrease with age, being mature by 33 weeks CA. PMID- 15807051 TI - Perspectives in palliative care. AB - OBJECTIVES: To review the evolution of palliative care in the United States and to define the terms palliative, supportive, and end-of-life care. DATA SOURCES: Published articles, textbooks, and reports. CONCLUSION: The health care demands that will arise from our aging society will force change in the field of palliative care. IMPLICATIONS FOR NURSING PRACTICE: Nurses providing care to patients who are living and dying from advanced cancer should apply palliative interventions from diagnosis until death. PMID- 15807052 TI - Communication issues and advance care planning. AB - OBJECTIVES: To provide practical strategies to enhance the quality of communication in the palliative and end-of-life medical care settings. DATA SOURCES: Published articles, textbooks, and reports. CONCLUSION: The components of effective and compassionate care at the end of life require successful communication with patients, families, and members of the health care team. Unfortunately, few health care professionals are formally trained in communication skills. IMPLICATIONS FOR NURSING PRACTICE: Nurses who possess self awareness and are skilled in effective communication practices are integral to the provision of high-quality palliative care for patients and families coping with advanced malignancies. PMID- 15807053 TI - Symptom clusters in advanced illness. AB - OBJECTIVES: To review the four sets of symptom clusters commonly seen in patients with advanced illness, and their definitions, associated symptomatology, and management. DATA SOURCES: Research and review articles and textbooks. CONCLUSION: Symptoms of patients with advanced illness tend to occur not isolation, but in symptom clusters. The ability to cluster symptoms in both assessment and management reduces the use of polypharmacy, systemic toxicities, and improves the patient's quality of life. IMPLICATIONS FOR NURSING PRACTICE: It is important that the nurse providing symptom management for the oncology patient understand the importance of clustering certain symptoms together. PMID- 15807054 TI - Pharmacology of palliative medicine. AB - OBJECTIVE: To describe the concepts of both pharmacokinetics and pharmacodynamics and how it applies to palliative medicine. DATA SOURCES: Research and review articles and textbooks. CONCLUSION: Most patients who have an illness that requires palliation are elderly, have co-morbid conditions, and are on many medications. Both age and disease can alter the way drugs are handled by the body (pharmacokinetics) and how they affect the body (pharmacodynamics). IMPLICATIONS FOR NURSING PRACTICE: The oncology nurse, armed with the correct knowledge about medications, can have an important inmpact on improving the patient's care, safety, and quality of life. PMID- 15807055 TI - Applying evidence to palliative care. AB - OBJECTIVES: To discuss the evidence-based practice process for palliative care, from defining a clinical problem to implementation and evaluation of an evidence based practice change. DATA SOURCES: Professional experience, literature, practice guidelines, and web sites related to evidence-based practice and palliative care. CONCLUSION: Nurses caring for patients needing palliative care need evidence from multiple sources. Using a systematic process to find and appraise evidence followed by strategically planning for implementation and evaluation of evidence-based practice changes optimizes the likelihood of enhanced patient care. IMPLICATIONS FOR NURSING PRACTICE: The integration of evidence-based practices into palliative care nursing will empower nurses to promote optimal patient outcomes and decrease practice variability. PMID- 15807056 TI - Cultural issues in palliative care. AB - OBJECTIVES: To review the common issues at the end of life across cultures and the measures that can be taken to dress these issues. DATA SOURCES: Published articles and textbook chapters. CONCLUSION: Culture plays a critical role in how patients, families, and health care providers view the end of life. Cultural competence in end-of-life care includes knowledge and experience and working in cross-cultural situations. IMPLICATIONS FOR NURSING PRACTICE: Increased culture specific knowledge will provide nurses with a basis for beginning exploration of individual or family beliefs. PMID- 15807057 TI - End-of-life care for children and adolescents. AB - OBJECTIVES: To describe the complexities in end-of-life care of children and adolescents dying cancer-related deaths. DATA SOURCES: Research studies, review articles, and government reports. CONCLUSION: The complexities in providing competent and compassionate care to the dying child and the family is intense, undeniable, and may in some clinical situations be unavoidable. IMPLICATIONS FOR NURSING PRACTICE: It is important for all those involved with the care at the end of life for a child or adolescent dying a cancer-related death create the framework that will support the provikion of competent and compassionate end-of life care. PMID- 15807058 TI - Ethical issues in palliative care. AB - OBJECTIVES: To review important issues that address respect for patient autonomy, beneficnce, non-maleficence, and justice, which are included in communication surrounding the determination of decision-making capacity, informed consent, breaking bad news, and creating shared goals of care. DATA SOURCES: Review articles, and government and organizational reports. CONCLUSION: Palliative care and its proximity to end-of-life care issues frequently raises ethical issues for patients, their families, and the clinicians caring for them. Supporting the identification and honoring the patient's preferences for treatment are central components of ethical behavior. IMPLICATIONS FOR NURSING PRACTICE: Advance care planning provides an important opportunity for respecting patient autonomy and may be helpful when discussing care options surrounding resuscitation, withholding or withdrawal of treatment, or the determination of medical futility. PMID- 15807059 TI - Modern palliative care: an exercise in prevention and partnership. AB - Palliative care is no longer solely an excercise in controlling suffering at the end of life. If we are to apply the principles of palliative care throughout the course of illness, both the culture and the organization of palliative care services must change. New partners must be enlisted in more sophisticated, increasingly academic partnerships. This change, while welcome and necessary, must come about with perservation of the core values of palliative care. PMID- 15807060 TI - Preparing for four challenges ahead. PMID- 15807061 TI - Marketing seniors' housing. PMID- 15807062 TI - The bishops and housing. AB - According to Catholic social teaching, housing is not a commodity but a human right. To ensure that all people--especially low-income elderly and other vulnerable populations--have access to affordable housing, the church has established a variety of programs, services, and advocacy efforts. Much of this work is based on key concepts: preserving existing housing stock, creating new programs to provide more options for the underserved, empowering residents and communities to deal with housing issues, establishing partnerships to make organizations' efforts more successful, making housing affordable, and ending discrimination in housing. Although church ministries, community groups, the private sector, and other players must work together to find solutions to the housing crisis, federal leadership is essential. Especially with the housing affordability gap growing and the U.S. population aging, the federal government must provide the resources, leadership, and direction for effective housing solutions. PMID- 15807063 TI - Allying health care and housing. AB - There is a wealth of evidence that health is inextricably linked to housing. For instance, research has shown that those in substandard housing have poorer health outcomes than other groups, and they often must forgo costly medication in order to pay for housing. Further, the health care and housing concerns faced by the underserved often compound one another--people with poor health often have trouble maintaining housing, and those with substandard homes, in turn, often have trouble maintaining their health. Three groups are especially vulnerable to the health care risks associated with housing issues: children, seniors, and the chronically homeless. As the research suggests, substandard housing is a contributing factor to the U.S. health care crisis. Therefore, as part of its efforts to reform the nation's health care system, the ministry should address housing issues as well. Seven Catholic health systems are doing this through the Strategic Health Care Partnership. The partnership, in collaboration with Mercy Housing, enables the seven organizations to work together to create healthy communities. The partnership's key goal is to increase access to affordable housing and health care. Just providing homes often is not enough, however. A holistic approach, through which supportive services are offered to the underserved, is most effective. PMID- 15807064 TI - Health, housing, and public policy. AB - With federal funding of affordable housing declining, health care and housing organizations must work together to advocate sound policy and reasonable funding in this realm. Federal agencies like the U.S. Department of Housing and Urban Development (HUD) and the U.S. Department of Agriculture traditionally have been the primary source of low income housing funds. But key housing programs like HUD's Section 8 have lost a significant amount of funding. Through advocacy efforts, health care and housing organizations can urge legislators to retain or restore these vital programs. They also can support the preservation of affordable housing units in order to counterbalance the trend of these homes being "lost to the market." Also, health care and housing agencies can partner to enhance housing services. Vulnerable populations-such as the elderly, individuals at risk for homelessness, those with disabilities, and the mentally ill-can benefit greatly from the supportive services that health care organizations can offer. PMID- 15807065 TI - Promoting health, building community. AB - As part of its mission to honor human dignity and to care for the poor and vulnerable, Catholic Healthcare Partners (CHP), Cincinnati, has made a systemwide commitment to address housing needs in the communities it serves. A priority for the system is providing safe, affordable housing options for the low-income elderly. CHP's approach goes beyond "bricks and mortar," however. The system aims not only to provide a home for senior adults but also to enrich their lives. Through various activities and support services, CHP's senior living complexes in Kentucky, Ohio, Pennsylvania, and Tennessee offer residents an opportunity to live in a vibrant community. CHP facilities have developed a variety of initiatives to enhance residents' lives. Among these are: spiritual care services, nurses who serve as a resource to low-income elders, a short-stay shelter for seniors in transition, a service referral program, and therapy to help elders remain independent. In order to offer these comprehensive services to senior adults, CHP relies on partnerships with a variety of organizations and on funding from both the federal government and private investors. Especially as the nation's population ages, CHP continues to make its housing ministry a strategic priority. PMID- 15807066 TI - Supportive housing in Brooklyn. AB - Since 1977, Saint Vincent Catholic Medical Centers of New York City has been creating and maintaining supportive housing offerings for at-risk populations, such as individuals with HIV/AIDS, those with substance abuse challenges, and the mentally ill. By providing a continuum of medical and social services, the organization aims to help residents stabilize and rebuild their lives. Saint Vincent sees empowerment as a key step toward helping individuals maintain their health, re-enter the community, seek employment, and pursue other goals. Some of the supportive services Saint Vincent offers in its housing communities are care coordination, counseling, peer support networks, self-help groups, leisure activities, help with finances, and referrals to community agencies offering other resources. Recognizing the importance of job training and assistance, the system also offers a unique program in which mentally ill individuals are able to work in a recycling center or office cleaning business, both of which are owned by Saint Vincent. PMID- 15807067 TI - A project in Pennsylvania. AB - Through collaboration, Mercy Health Partners (MHP) of Southwest Ohio, Cincinnati, and Catholic Social Services (CSS) of the Diocese of Scranton, PA, built a housing complex for low-income seniors and developed a variety of supportive services for residents. St. Catherine's Manor aims to meet elder adults' social and medical needs through offerings such as on-site health assessments, meal services, and transportation help. In order for the collaboration to be a success, MHP, CSS, and their partners had to trust one another. They also had to be clear and deliberate about their roles and responsibilities, thereby establishing expectations well in advance of the project. Their partnership was enhanced because they shared a common culture. PMID- 15807068 TI - Affordable housing in Southern California. AB - St. Joseph Health System (SJHS), Orange, CA, has been committed since the mid 1990s to acting as an advocate for the housing needs of low-income people. In 1997, for example, SJHS became a founder, with Mercy Housing and six other Catholic health care organizations, in creating the Strategic Health Care Partnership, an investment pool designed to bring affordable housing to the most vulnerable. In 1999, SJHS supported Mercy Housing's construction of an 81-unit development for very limited-income senior citizens in Orange County, CA. Two years later, SJHS supported Mercy Housing's creation of a 23-unit development in the county for homeless people with HIV/AIDS. In the same year, SJHS and other local groups formed the Kennedy Commission, a group devoted to improving housing in Orange County. Just last spring, the commission persuaded the city council of Anaheim, CA, to vote unanimously for an ordinance that would offer incentives to developers who set aside portions of their housing projects for low-income renters. PMID- 15807069 TI - Health and housing in Savannah. AB - St. Joseph's/Candler, a two-hospital system in Savannah, GA, and Mercy Housing East (MHE), Atlanta, a regional subsidiary of the Denver organization, collaborate on housing projects in southeastern Georgia. In Savannah, the partners transformed two local landmarks, a former hospital and a former school, into an 88-apartment development for low-income, predominantly African-American residents. St. Joseph's/Candler also provides health services to the community. As a result of their work, MHE and St. Joseph's/Candler have helped restore a historic city neighborhood. In Pembroke, GA, MHE (with support from St. Joseph's/Candler) has constructed a two-story, 30-unit, independent-living apartment development for senior citizens. Last year, St. Joseph's/Candler opened a health care facility in Pembroke. PMID- 15807070 TI - It takes a village. AB - Faced with the question of what to do with an aging, under-used hospital in northeast Detroit, St. John Health, Warren, MI, turned to the community for help in developing a plan for transforming the campus in a way that would better meet local needs. Through meetings and interviews with local residents, St. John Health learned that community members needed job training, career development, employment, health care, housing, and a safe environment. To meet these needs, St. John Health developed a plan to convert the facility, ST. John NorthEast Community Hospital, into a senior housing community called "Conner Creek: A partnership for healthy lives." Inpatient care has been phased out. Now St. John Health is working with local partners to build affordable, safe, and attractive apartments for seniors. Plans call for Conner creek to offer a wide variety of supportive services to complement the housing and to meet needs identified by the community. a nursing school, chronic care services, job training, and recreational activity areas are among the offerings under consideration. Townhouses may be added in the future. St. John Health's leaders say that Conner Creek May prove to a model for urban health care sites in need of revitalization. PMID- 15807071 TI - Catholic health care's future: Catholic Healthscan 2004" reveals the views of 175 ministry leaders. PMID- 15807072 TI - Journey into sponsorship's future. Trinity Health's public juridic person develops a formation program for prospective members. PMID- 15807073 TI - Avian influenza, Viet Nam--update. PMID- 15807074 TI - Shigellosis: disease burden, epidemiology and case management. PMID- 15807075 TI - Toward a biopsychosocial model for 21st-century genetics. AB - Advances in genomic research are increasingly identifying genetic components in major health and mental health disorders. This article presents a Family System Genetic Illness model to address the psychosocial challenges of genomic conditions for patients and their families, and to help organize this complex biopsychosocial landscape for clinical practice and research. This model clusters genomic disorders based on key characteristics that define types of disorders with similar patterns of psychosocial demands over time. Key disease variables include the likelihood of developing a disorder based on specific genetic mutations, overall clinical severity, timing of clinical onset in the life cycle, and whether effective treatment interventions exist to alter disease onset and/or progression. For disorders in which carrier, predictive, or presymptomatic testing is available, core nonsymptomatic time phases with salient developmental challenges are described pre- and post-testing, including a long-term adaptation phase. The FSGI model builds on Rolland's Family System Illness model, which identifies psychosocial types and phases of chronic disorders after clinical onset. The FSGI model is designed to be flexible and responsive to future discoveries in genomic research. Its utility is discussed for research, preventive screening, family assessment, treatment planning, and service delivery in a wide range of healthcare settings. PMID- 15807076 TI - The psychotherapy of genetics. AB - The evolution of genomic science and its effect on medicine and health care offer opportunities for family therapists to participate in the comprehensive care of patients and families with genetic disorders. This article provides an overview of what we now know about the psychological and interpersonal experience of patients and families facing some of these illnesses. Case examples illustrate the process of decision-making about testing and treatment, and the importance of understanding developmental issues and transgenerational family dynamics in any related psychotherapy. Challenging emotional issues include managing anger, ambivalence, and guilt; challenging interpersonal issues include dealing with differing coping and communication styles, decisions about disclosure and secrets, and conflict resolution. Family-oriented interventions include individual, couple, and family therapy, and psychoeducational groups. Recommendations are made for family therapists to participate as part of the genetic healthcare team. PMID- 15807077 TI - Transparency and self-disclosure in family therapy: dangers and possibilities. AB - Therapy is a paid intimate relationship that thrusts clients and therapists into navigating personal and professional boundaries. When, where, why, and how is it appropriate and ethical for family therapists to be transparent, and when is it damaging? Theorists take varied stances from Haley's position of tight boundaries around therapist disclosure--whether in treatment or training--to the narrative viewpoint that therapists should be transparent about models of therapy, personal values, and life experiences that inform their practice and beliefs. However, these positions are not research based, and theorists who support disclosure offer few guidelines other than general statements. This article examines the history of ideas about disclosure in six major family therapy models, and the dangers and possibilities of transparency. It looks at the research on self disclosure in individual therapy and whether and how it could apply to family therapy. Guidelines are proposed that take into account the multiple social identities of therapists and clients, and issues of safety and transparency. PMID- 15807078 TI - Structured methods and striking moments: using question sequences in "living" ways. AB - This article draws together two seemingly incompatible practices in social constructionist therapies: the use of structured questioning methods (associated with solution-focused and narrative therapies) and the poetic elaboration of "striking moments" (associated with conversational therapies). To what extent can we value and use both styles of practice? Beginning with practitioners' concerns about the use of structured question sequences, I explore possibilities for resituating these methods in different conceptual and metaphorical frames, selectively drawing on ideas from the philosophy of striking moments. The aim is not to reduce one therapeutic style to another, but to encourage the teaching and practice of structured methods in more creative, improvisational, and "living" ways. PMID- 15807079 TI - Family ties: constructing family time in low-income families. AB - "Family time" is reflected in the process of building and fortifying family relationships. Whereas such time, free of obligatory work, school, and family maintenance activities, is purchased by many families using discretionary income, we explore how low-income mothers make time for and give meaning to focused engagement and relationship development with their children within time constraints idiosyncratic to being poor and relying on welfare. Longitudinal ethnographic data from 61 low-income African American, European American, and Latina American mothers were analyzed to understand how mothers construct family time during daily activities such as talking, play, and meals. We also identify unique cultural factors that shape family time for low-income families, such as changing temporal orientations, centrality of television time, and emotional burdens due to poverty. Implications for family therapy are also discussed. PMID- 15807080 TI - Maternal depression, adult attachment, and children's emotional distress. AB - INTRODUCTION: Our goal was to evaluate the impact of depressed mothers' marital intimacy and attachment security in romantic relationships on children's internalizing and externalizing symptoms. METHOD: Forty-six clinically depressed mothers rated attachment security, marital intimacy, and symptoms shown by their children aged 8 to 12. RESULTS: Maternal avoidance of closeness predicted increases in children's internalizing symptoms over a 6-month period. DISCUSSION: Avoidance of closeness in depressed mothers may be implicated in the development of internalizing symptoms in their children, possibly because individuals who are avoidant of closeness are poor caregivers. PMID- 15807081 TI - Critical consciousness, accountability, and empowerment: key processes for helping families heal. AB - Families are inextricably embedded within their larger sociopolitical contexts, an observation acknowledged by many theorists. The field of family therapy is working on its translation into comprehensive and effective approaches for helping families change. This article illustrates the use of the Cultural Context Model to help families change, guided by the linked foundational concepts of accountability, critical consciousness, and empowerment. The authors support their discussion of theory with examples illustrating the ways in which social patterns connected to race, gender, class, and sexual orientation shape the dilemmas that family members encounter, as well as their access to solutions. PMID- 15807082 TI - Co-constructing a cross-cultural course: resisting and replicating colonizing practices. AB - This article examines the cross-cultural complexities of teaching and learning processes at different levels in a family therapy training program delivered in a different country. It presents two interlinked narratives of an advanced family therapy training program delivered in Slovenia by a U.K-based family therapy training institute. The context in which the training was negotiated and planned is discussed, highlighting issues at the macrosystemic level. Dilemmas that arose in relation to differing beliefs about teaching and learning are elaborated. We examine power relationships, ways that we positioned ourselves and each other, and how we tried and sometimes failed to take these into account. We question how the issue of spoken language and its meanings creates its own constraints and power structures, and how we respond to the impossibilities of translation. We argue that it is through the examination of uncomfortable processes that we develop self-reflexivity and work toward more equitable relationships. PMID- 15807083 TI - Shame, guilt, self-hatred and remorse in the psychotherapy of Vietnam combat veterans who committed atrocities. AB - The soldiers who perpetrated atrocities during the Vietnam War are a group much less studied than the victims of violence during peacetime. These soldiers were catalyzed by war to express a darker side of their humanity, a side that our society often chosses to ignore. At some level their experiences relate to us all. In the future more young men and, probably young women, will be sent to war. Once they have been transformed by military training and the hellish conditions of war, society tends to forget them--to leave them wandering in a no man's land of their own tortured thoughts and feelings. We need to understand more about how such transformations of the self beset our returning soldiers. And, once this has happened, we need to know how to help them reintegrate into society and reconnect with others in a meaningful way. A psychotherpy that embraces the patient's need to express remorse is necessary in order to help these patients work through their guilt and self-hatred. These are the main objectives of treatment. PMID- 15807084 TI - Is psychotherapy good for your health? AB - This is a dedicated review of the evidence for the relation of having a period of psychotherapy and then comparing it with a measure of improved physical health. We aimed to make it the first intended-to-be-complete review of this type. Three inter-related types of studies were examined: Type 1: reduction in physical illnesses through psychotherapy, especially for the patient's survival time during the interval between diagnosis and an end point, Type 2: reduction in pain in relation to receiving psychotherapy, and Type 3: reduction in costs of treatment in relation to receiving psychotherapy. To find the relevant studies on these topics, we performed a literature search using both Psychinfo and Medline databases. An average of the effect sizes under each type was taken to calculate the mean effect size along with its confidence interval. Our results (1) on survival time for the combined severe patients, did not reach even the lowest significant level of effect size, although the low severity patients seemed to fit the hypothesis better, but the other two reduction topics, (2) and (3), clearly did achieve it. PMID- 15807085 TI - The silence of Socrates. AB - This paper emphasizes the humanistic and phenomenologic approach to the mind (soul, self) and contrasts it to the "era of the brain" that is so idealized by psychiatry today. It describes the debate between those who expect science to explain everything sooner or later and those who believe there are certain essential aspects of the world, such as the qualia of consciousness, that cannot be reduced to material factors. Plato's dialogue Timaeus is taken as the historically first example of this debate. The connection between it and what has been labeled "neuroism" in contemporary psychiatry is established and the relevance of this debate to the practice of psychoanalysis and dynamic psychiatry is discussed. Finally the wider consequences of the scientific or materialistic approach to the world as forecast already by Nietzsche are indicated. PMID- 15807086 TI - Personality disorders over time: implications for psychotherapy. AB - Personality disorders have an early onset, and are associated with dysfunction over the course of adult life. Antisocial and borderline personality disorders tend to remit with age, but other categories do not usually show improvement. The chronicity of personality disorders is both a challenge and a frame for treatment planning. Psychotherapy for these patients can focus on rehabilitation and the development of social niches that match their personality profiles. PMID- 15807087 TI - Psychotherapy with physically disabled patients. AB - With the last decades, health care interventions have been more productively attuned to actualizing the potential for optimal recuperation of every patient. Unique and important contributions of psychotherapy to this effort include: 1) A formulation which synthesizes an understanding of clinical behaviors, reality based physical limitations and risks with an appreciation of the patient's mechanisms of defense, ego strengths and weaknesses, and transference expectations which impact the treatment process; 2) The utilization of individual psychotherapy (focused on "insight") in combination with supportive individual and group experiences. For children and adolescents struggling with age appropriate physical-developmental and social issues or learning disabilities, psychoeducational approach for disabled youngsters has proven very beneficial. 3) Occasional crises occur which involve the spouse or relatives more than the index patient. Working to provide supportive Couple or Family System intervention is sometimes as essential as caring for the disabled individual. 4) Numerous Group Therapy approaches have proven efficacious. Treatment in a group setting is attractive to those who are concerned about cost-containment. Unfortunately, groups for disabled are often "didactic" and utilize a format that provides factual information about disabilities, medical procedures, and sometimes an intellectual discussion of "emotional answers" for certain types of problems or conditions. Groups that facilitate self-disclosure and emotional interactions among the members accomplish more meaningful results. In conclusion, we wish to emphasize the importance of developing rigorous scientific research in the area of disabilities which will match the excellence of clinical work already being done in the field. Gaining an accurate and more thorough understanding of the psychological reality of a disabled person's internal world may be a key to facilitating his or her self-esteem and optimal adaptive functioning. PMID- 15807088 TI - Schiavo case's fallout. Providers concerned about political intrusion! PMID- 15807089 TI - Priceless. Healthcare consumers show little interest in chargemasters despite growing movement to mandate price disclosure. AB - More states are joining the trend of requiring hospitals to publicly report prices, with South Dakota becoming the latest to sign on. But are consumers taking advantage of the service or having any luck understanding the complex science of hospital billing? "I would say it's impossible," says Reece Fawley, left, adding that only a handful of people have requested a look at his hospital's chargemaster. PMID- 15807090 TI - Seeing red. California hospitals vulnerable to bankruptcy. PMID- 15807091 TI - Reaching out. Physician investors hope to save Atlanta hospital. PMID- 15807092 TI - Marketing switch. Brailer's office was seeking IT success stories. PMID- 15807093 TI - Masimo wins new round. Tyco unit plans appeal of dollar 420 million antitrust verdict. PMID- 15807094 TI - A change of plans. Beverly agrees to auction after hostile takeover moves. PMID- 15807095 TI - Backpedaling on safety. Demand for confidentiality only another example of whitewashing problem. PMID- 15807096 TI - Time to reform 990s? Advocates say IRS filings could use an overhaul to improve comparability of executive compensation. PMID- 15807097 TI - Ensuring care. Minimal insurance coverage has pros, cons. PMID- 15807098 TI - Battling the system. Commonwealth Fund chief fights for quality, transparency. Interview by Michael Romano. PMID- 15807099 TI - Modulation of the gene expression in the protective effects of electroacupuncture against cerebral ischemia: a cDNA microarray study. AB - In the present study, a cDNA microarray technology was used to analyze the gene expression profile in ischemia and in electroacupuncture (EA)-treated ischemia. A focal cerebral ischemia/reperfusion model on Macaca mulatta was performed with a modified middle cerebral artery occlusion method. Among the nearly 8000 genes, approximately 8% of the total number of genes examined were affected after ischemia/reperfusion injury. Major altered genes were downregulated. In EA treated monkeys, approximately 10% of the total number of genes examined were affected. Major altered genes were upregulated, including signal transduction-, cell-cycle-, metabolism-, stress response-, DNA repair-related genes. One of the representative upregulated genes encodes insulin-like growth factor-1 (IGF-1) was confirmed using in situ hybridization. Results showed that after ischemia/reperfusion injury, IGF-1 mRNA expression decreased in ipsilateral striatum, whereas increased in ipsilateral hippocampus. No expression changes were observed in cortex. EA treatment could obviously upregulate the IGF-1 mRNA expression in striatum, and further enhance its expression in hippocampus. Therefore, the data presented suggest a possible mediator underlying the mechanisms of anti-ischemic effect of acupuncture. In conclusion, the protective mechanisms of EA against stroke include several related pathways and gene expressions. Microarray analysis may provide a framework for understanding these complicated mechanisms and yield valuable, clinically relevant insights and potentially therapeutic targets of stroke. PMID- 15807100 TI - Managing acute withdrawal syndrome on patients with heroin and morphine addiction by acupuncture therapy. AB - Though there are articles and case reports about using acupuncture to detoxify and to break the narcotic addiction, few articles describe in the West about using acupuncture therapy to treat the emergence of acute withdrawal symptom due to heroin, opium, or morphine. Most often the method of treatment are using the methadone or benzodiazepine and phenoziazine drugs this article describes many years of clinical experience with non-drug approach to treat the acute withdrawal symptoms with acupuncture therapy. Unlike the drug approach, which usually has side effects, there is no adverse effect with acupuncture therapy. PMID- 15807101 TI - Left-right and Yin-Yang balance of biophoton emission from hands. AB - Yearlong measurements of biophotons from palm and back of hand of three healthy people were performed. The detection of biophoton was done with two photomultiplier tubes whose spectral range was from 300 nm to 650 nm. The measurement was done on a circular area of diameter 46 mm whose centers were at the acupuncture point Laogong (PC8) of a palm and the mid-point of a back, respectively. The emission rates from the dorsa showed strong seasonal dependence which is consistent with the active nature of Yang meridians, while the palm sides show less seasonal dependence as they belong to the passive Yin meridians. This could be quantified simply by the standard deviations from the yearly average: They were 47.6 counts per second (cps), 66.1 cps and 66.0 cps from the dorsa, and 23.8 cps, 29.7 cps and 30.4 cps from the palms of subject 1, 2 and 3, respectively. Biophoton emission revealed qualitative individual tendencies: The subjects 1 and 2 emitted more strongly from the left dorsa, while the subject 3 from the right dorsum. The left-right balance of biophoton emission was well kept for normal people, which is in contrast with the severe imbalance for some stroke patients. Thus biophoton measurement has a diagnostic potential that encourages more studies. PMID- 15807102 TI - Registering the response of tissues exposed to an interferential electric current stimulation. AB - Electrotherapy is a treatment method in which an electric stimulus is used to achieve physiological changes in the body. When increasing the level of the electric stimulus, three sensory, three motor and a pain response can be found. In this paper the three sensory threshold levels (1st just noticeable weak tingling sensation, 2nd a very fine but definite tingling and 3rd moderate strong tingling) were studied with an interferential electric current stimulus. The interferential electric current can be achieved by summing two alternating currents with little different frequencies with each other. As a current source, the study used a value intended for interferential electric current therapy. The threshold value measurements were performed on 5 days to 6 volunteers. The method of limits is a frequently used technique for determining sensory thresholds where ascending or descending stimuli is used and the observer reports of the appearance or disappearance of the sensation, respectively. A Bekesy's algorithm is a threshold tracking method with a variable stimulus. They were used for controlling the interferential electric current stimulus, and hence, for studying the,thresholds. By using the method of limits, the measurements were done at three sensory threshold levels at interferential frequencies deltaf of 10 Hz, 30 Hz, 50 Hz, and 100 Hz caused by two sine waves, one with a frequency between 2490 Hz and 2400 Hz, and the other constant 2500 Hz. In the method based on Bekesy's algorithm the interferential frequency deltaf was increased from 1 Hz to 100 Hz by changing the first frequency from 2499 Hz to 2400 Hz and the measurement was repeated for all three sensory thresholds. The sensory thresholds for interferential current stimuli seem to be quite unaffected by the interferential electric current frequency among the healthy test persons, and no distinct interferential electric current frequencies at which the threshold values should be measured were found in this study. PMID- 15807103 TI - An examination of the relationship between five oriental musical tones and corresponding internal organs and meridians. AB - The Yellow Emperor's Internal Medicine (Nei-Jing) describes the relationship between five internal organs corresponding to five different meridians and five oriental musical tones such as Gung (spleen), Sang (liver), Gak (liver), Chi (heart), Wu (kidney). However, there has been little research to test this relationship. The purpose of this study was to determine this relationship through applying sound stimulation for 60 seconds, comprising various frequencies produced on an electric organ(Gung: 261.6 Hz; Sang: 293.7 Hz; Gak: 329.6 Hz Chi: 392.0 Hz and Wu: 440.0 Hz), to 20 healthy adults by stimulating their ears (via headphones), skin (via speakers) and both. A significant relationship was found between the tones and the meridians as they are described in Nei-Jing. The greatest response rates were found in the following corresponding organic and musical tones: spleen/Gung, lung/Sang, liver/Gak, heart/Chi, and kidney/Wu. The traditional descriptions of Yellow Emperor's Nei-Jing are therefore consistent with the findings of this study. Further basic research may lead to the application of clinically appropriate tones in an acoustically suitable room following oriental medical diagnosis. PMID- 15807104 TI - [Preparation of covalently bonded cellulose tris (4-methylbenzoate) derivative chiral stationary phases through a polymerization reaction]. AB - Cellulose tris (4-methylbenzoate) derivatives (CTMB) having methacryloyl groups were synthesized with regio-selective or nonselective procedures, and were immobilized on gamma-methacrylatepropylated silica (gamma-MAPS) through a polymerization reaction. The obtained chiral stationary phases ( CSPs) were evaluated by high performance liquid chromatographic (HPLC) resolution of 9 racemates (3-butyl-phthalide, ketoprofen, trans-stilbene oxide, benzoin, benzoin analogue, Troger' s base, warfarin, 4,4'-dimethoxy-5,6,5',6'-dimethylenedioxy biphenyl-2,2'-dicarboxylate and 4,4'-dimethoxy-5,6,5',6'-dimethylenedioxy biphenyl-2-methylcarboxylate-2'-ethylcarboxylate). For regio-nonselective procedure, increasing the content of vinyl group on CTMB would lead to higher immobilization efficiency and enantio-selectivity. CSPs prepared by regio selective procedure possessed slightly higher enantioselectivities than those prepared by regio-nonselective procedure, while the latter showed some advantages for rapid and facile preparation. CSPs prepared with silica gel II (Fuji, 5 microm, 30 nm, ca. 150 m2/g) showed higher enantioselectivities than those prepared with silica gel I (Kromasil, 5 microm, 20 nm, ca. 240 m2/g). The prepared CSPs could keep stable under different eluents, even with 40% of tetrahydrofuran (THF) in mobile phases. PMID- 15807105 TI - [Determination of neurosteroids in rat brain regions by liquid chromatography/negative atomspheric pressure ionization mass spectrometry]. AB - A simplified method has been established using liquid chromatography-negative atomspheric pressure ionization mass spectrometry (LC-MS) to simultaneously determine two conjugated neurosteroids from rat brain regions. Neurosteroids were separately isolated in a two-step procedure using chloroform/2-butanol (50: 50, v/v) where the first step was to extract sulfated steroids, then steroid fractions were purified by solid phase extraction (SPE), and finally the sulfated steroid was solvolyzed. Estrogen sulfate was chosen as internal standard. All steroids were derivatized with 2-nitro-4-trifluoromethylphenylhydrazine (2NFPH) and analyzed by LC-MS using selected-ion monitoring. LC-MS was performed on an Agilent 1100 liquid chromatograph-mass spectrometer with atmospheric pressure chemical ionization (APCI) interface, and the hydrazones of oxosteroids was analyzed in a negative-ion mode. A Zorbax SB C18 column was used with a flow rate of 1 mL/min at 40 degrees C. The mobile phase consisted of acetonitrile and distilled water. The concentrations of PREGS and DHEAS in male rat brain regions were (4.14 +/- 1.33 ) ng/g and (2.26 +/- 0.76) ng/g (pituitary gland), (1.98 +/- 1.13) ng/g and (1.80 +/- 0.93) ng/g (hypothalamus), (1.08 +/- 0.48 ) ng/g and (0.81 +/- 0.23) ng/g (frontal cortex), (0.72 +/- 0.19) ng/g and (0.77 +/- 0.12) ng/g (hippocampus), (1.70 +/- 0.45) ng/g and (1.44 +/- 0.71 ) ng/g (amygdale), (0.92 +/- 0.27) ng/g and (0.85 +/- 0.44) ng/g (striatum), (3.62 +/- 1.77) ng/g and (3.17 +/- 2.11) ng/g (nucleus accumbens), respectively. Good linearity and accuracy were observed for each steroid. The procedure was suitable for measuring concentrations of the sulfated steroids in rat brain regions simultaneously. PMID- 15807106 TI - [Determination of bisphenol A and 4-nonylphenol in rat tissues by high performance liquid chromatography with fluorescence detection]. AB - A simple and highly sensitive method has been developed for the determination of bisphenol A (BPA) and 4-nonylphenol (4-NP) in rat tissues by high performance liquid chromatography (HPLC) with fluorescence detection. The rat tissue samples were homogenized-with methanol-ammonium acetate buffer (2:8, v/v) by a high-speed homogenizer and then BPA and 4-NP were extracted by a mixed solvent of n-hexane and diethyl ether (7:3, v/v). The organic layer was evaporated with a stream of nitrogen, and the residue was dissolved in the mobile phase. The chromatographic operating conditions were a C18 column (250 mm x 4. 6 mm i.d., 5 microm), acetonitrile-ammonium acetate buffer (pH 4.5) (75:25, v/v) as mobile phase at a flow rate of 1 mL/min, fluorescence detection with the excitation and emission wavelengths at 227 nm and 313 nm, respectively. Average recoveries for rat tissues at three different levels were in the range of 82.0% -95.4% for BPA and 81.2% -96.5% for 4-NP. The relative standard deviations (RSDs) were 0. 37% - 5.28% and the detection limits were 4.0, 4.6, 3.2, 3.3 ng/g for BPA and 12.0, 15.6, 11.8, 13.8 ng/g for 4-NP in liver, kidney, heart and brain tissues, respectively. The intra-day precisions were 0.89% -4.50%, and the inter-day precisions were 3.10% - 12.40%. These results demonstrated that the proposed method is simple, sensitive, and reliable for the determination of BPA and 4-NP in animal tissues. PMID- 15807107 TI - [Study on thermostability and photo-isomerization of trans-resveratrol by high performance liquid chromatography and liquid chromatography-electrospray ionization-mass spectrometry]. AB - The thermostability and photo-isomerization reaction mechanism of trans resveratrol were studied by using high performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization-mass spectrometry ( LC-ESI-MS). A reversed-phase Hypersil-ODS column was used with the mobile phase consisting of 60/40 methanol-water, with addition of 0.05% trifluoroacetic acid for HPLC, and 0.1% formic acid and 5 mmol/L ammonium formate for LC-ESI-MS, at a flow rate of 0.5 mL/min. Diode array detection was set at 300 nm and the injection volume was 20 or 10 microL. Mass spectrometric conditions were in the negative mode. The results of thermostability test under 4, 20 and 40 degrees C indicated that the concentration of trans-resveratrol, after storage for 600 h, was decreased slightly under 4 degrees C, decreased by 5% under 20 degrees C, and 10% under 40 degrees C. In addition, the photo-isomerization reaction of trans- and cis resveratrol under 254 nm and 365 nm UV irradiation was investigated by LC-ESI-MS, showing that the isomerization is based on p-status, and that some secondary reactions are radical induced. PMID- 15807108 TI - [Characterization method for metallothioneins by reversed-phase liquid chromatography coupled with electrospray ionization mass spectrometry]. AB - Mammalian metallothioneins (MTs) usually have two major isoforms, MT-1 and MT-2, existing in tissues because of the genetic polymorphism. Metallothionein isoforms and sub-isoforms were characterized with a hyphenated technique of reversed-phase liquid chromatography (RPLC) coupled with electrospray ionization mass spectrometry (ESI-MS). The MT-isoform separation was carried out using a C8 narrow-bore column Vydac C8 (250 mm x 2.1 mm i.d., 5 microm, 30 nm). Buffer A was 5 mmol/L ammonium acetate in water (pH 6.0) and buffer B was 5 mmol/L ammonium acetate in methanol-water (1:1, v/v) (pH 6.0). A linear gradient elution was used by increasing buffer B from 10% to 37.5% within 40 min. The flow rate was set at 0.20 mL/min. The tentative formulas of MT isoforms and subisoforms were interpreted with the measured masses at the vicinity of apexes in total ion current chromatogram of ESI-MS and the reference data reported previously. The method was validated with the standard reference materials of MT-1 and MT-2. The results indicated that many MT species interpreted were in good agreement with the reference data reported. The MT sample from rabbit liver was measured with the established method. The results, of which the apo-MT was used by acidification of the sample for simplifying the mass spectra, showed that there were species of MT isoforms and subisoforms in rabbit livers. PMID- 15807109 TI - [Study on chiral selective inclusion and retention characteristics of cis-trans isomers and enantiomers of sertraline with HP-beta-CD as a mobile phase modifier]. AB - The effect of the inclusion complex formation of cis-trans isomers and enantiomers of sertraline with added hydroxypropyl-beta-cyclodextrin (HP-beta-CD) in the reversed-phase high performance liquid chromatographic (RP-HPLC) mobile phase on the retention and selectivity has been investigated. Graphs of 1/k vs [HP-beta-CD] in different experimental conditions gave good linear relationships, indicating the stoichiometry ratio of 1:1. The binding constants between HP-beta CD and cis-trans isomers and enantiomers of sertraline were calculated by using the liquid chromatographic retention values. Meanwhile, the influences of pH value, buffer ionic strength, concentration of organic modifier and temperature on the binding constant have been studied in detail. Quantitative equations were obtained for the correlations between the binding constants and the above influencing parameters. On the other hand, the thermodynamic parameters of the inclusion complexes were determined for the four inclusion complexes. The study characterized the chiral recognition selectivity between sertraline isomers and HP-beta-CD. It was demonstrated that the chiral selectivity values were in the same sequence as delta(j,i) deltaH and delta(j,i) deltaS and opposite order against the binding constants of inclusion complexes. Both enthalpic and entropic terms were negative for cis-trans isomers and enantiomers of sertraline. It was confirmed that enthalpy-driven separations occurred in consistency with the results. The influence of temperature on retention behavior showed that the process of inclusion was exothermic, in consistency with the calculated thermodynamic parameters. PMID- 15807110 TI - [Study of protein binding in ketoprofen using liquid chromatography frontal analysis in comparison with capillary electrophoresis frontal analysis]. AB - A method of high performance liquid chromatography-frontal analysis (HPLC-FA) was developed to study the binding of ketoprofen to human serum albumin (HSA) and it was compared with high performance capillary electrophoresis-frontal analysis (HPCE-FA). The separation was performed using Pinkerton GFF II-S5-80 internal surface reversed-phase silica column (150 mm x 4.6 mm i.d., 5 microm) at pH 7.4 in a 67 mmol/L isotonic sodium phosphate buffer at 37 degrees C. Other conditions included a flow rate of 0.2 mL/min, UV detection at a wave-length of 254 nm and an injection volume of 950 microL. A trapezoidal peak of the unbound ketoprofen appeared after HSA elution in the chromatogram. The plateau height of the peak was employed to determine the concentration of unbound ketoprofen in the HSA equilibrated sample solution. The HPLC-FA method provides the advantage of high sensitivity and however the disadvantages of large sample size and long analytical time when compared with HPCE-FA. HPLC-FA is applicable to the binding parameter estimation of ketoprofen to both primary and secondary sites, which are 0.37 x 10(6) L/mol and 1.4 for K1 (the association constant) and n (the number for the binding sites per molecule HSA), respectively, and 0.005 x 10(6) L/mol and 7.2 for K2 and n2, respectively. In contrast, HPCE-FA measures parameters for only the secondary binding sites; K2 of 0.018 x 10(6) L/mol and n2 of 2.54 can be estimated. It is found that ketoprofen binds mainly at the primary sites at a lower mole ratio of ketoprofen versus HSA, and the binding at the secondary sites occurs at a higher ratio. PMID- 15807111 TI - [Separation of tannins in Rhubarb and its analysis by high performance liquid chromatography-mass spectrometry]. AB - In order to investigate the pharmaceutical actions of rhubarb, a method for extracting, separating and analyzing the tannin components in rhubarb was studied. At first, a procedure for the group separation of tannins from the water ethanol extract of rhubarb was established based on the formation of tannins caffein precipitation. Then, a high performance liquid chromatographic (HPLC) method for the analysis of tannins in rhubarb was developed. This HPLC method is based on a reversed-phase C18 column and polar mobile-phase such as water and methanol with gradient elution, and the tannins can be well separated. Finally, the identification of the tannin components in rhubarb was carried out by high performance liquid chromatography-mass spectrometry (HPLC-MS). The structures of the main tannin components (gallic acid, catechin, the dimer, trimer, tetramer and pentamer of catechin) in rhubarb are suggested. The fragmentation laws of the tannin components are summarized. In comparing with previous methods, it is simple and without the interference of co-existed compounds. PMID- 15807112 TI - [Determination of curcumin in urine by capillary electrophoresis]. AB - The major component of the plant curcuma longa (a widely cultivated tropical plant in Asia and Central America) is curcumin. Curcumin has been reported to have very strong anti-inflammatory, anti-carcinogenic, anti-oxidant, antiallergic, anti-bacterial, and anti-tumor activities. Little is known about the absorption, distribution, and metabolism of curcumin in human beings. The first step in in vivo physiological and pharmacokinetic studies is to develop a method to measure curcumin in body fluid. A rapid capillary electrophoretic (CE) method with diode array detection was established for the determination of curcumin in human urine. It could be rapidly determined within 2.5 min. The optimized experimental conditions were as follows: 15 mmol/L Na2B4O7 as buffer, applied voltage 20 kV, temperature 25 degrees C and detection wavelength 262 nm. The method has been validated and shows good performance with respect to selectivity, reproducibility and limit of detection. Curcumin had good linearity in the range of 10 - 300 mg/L, and the recoveries of curcumin added in urine were more than 96.3% with relative standard deviations (RSDs) less than 2.3%. The method is sensitive, fast and accurate and can be used to determine curcumin in urine. PMID- 15807113 TI - [Design of laser-induced fluorescence detector for high performance liquid chromatography]. AB - Based on confocal configuration, a laser-induced fluorescence detector (LIFD) for high performance liquid chromatography (HPLC) is described and evaluated. A diode double-pumped solid-state laser emitting at 473 nm is used as excitation source. A detection cell of 7 microL physical volume and 0.1 nL effective optical volume was designed for conventional HPLC-LIFD. The angle between entrance and outlet channels of the cell was 120 degrees, without any change of internal diameters or right-angles along the channels. These measures insured the elimination of bubble accumulation or formation inside the cell, and additional band broadening effect. Riboflavin standard was used as test sample to evaluate the performance of the HPLC-LIFD. The analyses of real samples containing riboflavin were demonstrated. PMID- 15807114 TI - [Recent advances in chromatography coupled bioassay for screening bioactive natural products]. AB - Chromatography is the important platform for screening bioactive compounds. The recent advances in chromatographic technology of thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC) coupled to bioassay and its application are reviewed. Thirty references were cited. PMID- 15807115 TI - [Effect of emulsion on chromatographic process]. AB - The types and features of concentration wave of the chromatographic process in which the mobile phase was emulsion were studied theoretically by the method of characteristics and the concept of multi mobile phase chromatography. When the mobile phase is an emulsion, solute is partitioned between the inner and outer phases of the emulsion as well as simultaneously adsorbed on the stationary phase resulting in the great difference of concentration profiles from the case in which the mobile phase is homogenous. The various profiles and mechanisms of concentration wave of the chromatographic process in which the partition relationship between the inner and outer phases of the emulsion and the adsorption isotherm on the stationary phase were both Langmuir-like were analyzed and discussed with corresponding examples, and the comparison with the case in which the mobile phase is homogenous is also done. It is shown that the chromatographic progress with emulsion is more complicated and multiple. The existence of multi mobile phases may sharpen simple wave to be shock wave or diffuse shock wave to be simple wave. PMID- 15807116 TI - [Determination of the binding constants of four anti-HIV-1 active compounds with bovine serum albumin by capillary zone electrophoresis]. AB - The binding constants (Kb) of four novel anti-HIV-1 active compounds with bovine serum albumin (BSA) were determined by capillary zone electrophoresis (CZE) with UV detection under condition of phosphate buffer (pH 8.0, 50 mmol/L) at 15 kV running voltage. These molecules were synthesized by computer-simulated design with the property of inhibiting the binding of HIV-1 Tat protein to trans activation response region (TAR) RNA required for HIV-1 transcription and blocking the HIV replication cycle. The results showed that with the addition of different concentrations of BSA into the buffer solution, the binding constants of four active compounds (IG3, iso-C3, C3, MC3) with BSA could be measured by the change of migration time. The experimental values of Kb were 1.07 x 10(4), 1.34 x 10(4), 8.51 x 10(3) and 9.45 x 10(3) L/mol, respectively. It is an easy and simple method to estimate the interaction of small molecules with biomacromolecules with 1:1 molar binding ratio. PMID- 15807117 TI - [Enantioseparation of tropicamide by capillary electrophoresis with square wave amperometric detection]. AB - An enantioseparation method for tropicamide by high performance capillary electrophoresis with square wave amperometric detection (SWAD) was developed. The enantiomers of tropicamide were baseline separated in 16 min with an uncoated fused-silica capillary (75 microm i.d. x 50 cm) under the optimum conditions: 7 mmol/L Tris-10 mmol/L citric acid-2 mmol/L H3BO(3)-15 mmol/L beta-cyclodextrin (beta-CD) (pH 3.0) as background electrolyte, SWAD balance potential (E(b)) + 0.80 V (vs. Ag/AgCl), separation voltage 15 kV, injection height 20 cm, and injection time 10 s. The calibration curve of the enantiomers showed good linearity in the range from 5 micromol/L to 750 micromol/L with detection limit of 2 micromol/L. The average recoveries of added standards were 96% - 103%. The effects of the concentrations of beta-CD and the boric acid, the pH of the background electrolyte on resolutions (Rs) of the enantiomers were discussed in details. The proposed method was applied to the determination of a commercial tropicamide eye-drops sample without pre-treatment, and satisfactory results were obtained. PMID- 15807118 TI - [Preparation and evaluation of 2,6-di-O-pentyl-beta-cyclodextrin bonded silica stationary phase for high performance liquid chromatography]. AB - In order to improve the chiral separation capability of the conventional beta cyclodextrin bonded-silica gel stationary phase, 2,6-di-O-pentyl-beta cyclodextrin bonded stationary phase (PCDS) was prepared via a long spacer. The resulted bonded-silica stationary phase was characterized by three methods, namely Fourier transform infrared, Molisch color reaction, X-ray optical electrical energy spectrogram. The chromatographic performances of PCDS were investigated by using liquid chromatography with toluene, dimethyl phthalate, and phenanthrene as solutes, and their retention mechanism was investigated and discussed. The results show that the introduction of pentyl to beta-cyclodextrin leads to enhancement of the retention of the solutes. The chiral separation capability of the new bonded-silica stationary phase was evaluated by using liquid chromatography with some chiral drugs. Some of the enantiomers such as chlorphenamine maleate and bupropion hydrochloride were separated by heptakis (2,6-di-O-pentyl)-beta-cyclodextrin bonded silica stationary. PMID- 15807119 TI - [Simultaneous determination of ten sulfonylurea herbicide residues in soybeans by high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed for the simultaneous determination of ten sulfonylurea herbicide (oxasulfuron, thifensulfuron-methyl, metsulfuron-methyl, triasulfuron, chlorsulfuron, bensulfuron-methyl, prosulfuron, pyrazosulfuron-methyl, chlorimuron-ethyl and primisufuron-methyl) residues in soybeans. Sulfonylurea herbicides were extracted with acetonitrile, followed by hexane partitioning. After the extract was cleaned up with a Florisil column, sulfonylurea herbicides were analyzed by HPLC-photo diode array detector (DAD) and quantified by external standard method. The pre-treatment method of the samples and the chromatographic conditions of the analysis were critically examined. The linear ranges of 10 sulfonylureas were 0.1 - 10.0 mg/L, and the correlation coefficients were 0.999 6 - 0.999 7. The average recoveries of ten sulfonylurea herbicides in spiked soybeans ranged from 69.8% to 100.7%, and the relative standard deviations were between 1.89% and 10.43%. The limit of detection was 20 microg/kg. The results have indicated that the method developed is easier, faster, and has better purification effect. It has also demonstrated that this multiresidue analytical method can meet the requirements for the simultaneous determination of many sulfonylurea herbicides in import and export inspection for soybeans. PMID- 15807120 TI - [Determination of the content of geniposide in xinxue granules by high performance liquid chromatography]. AB - A high performance liquid chromatographic method has been developed for the determination of geniposide in Xinxue granules. Geniposide was extracted by ultrasonic extraction for 30 min. The chromatographic separation was carried out on a Diamonsil C18 column (200 mm x 4. 6 mm i.d., 5 microm) with a mobile phase consisting of an acetonitrile-water (15:85, v/v) mixture. The detection wavelength was set at 238 nm. The calibration curve was linear in the ranged of 25 - 400 mg/L for geniposide. The average recovery of geniposide was 101.2% with a relative standard deviation (RSD) of 1.6% and contents of geniposide in the granules ranged from 0.841 to 0.923 mg/g. This method is simple, reliable and suitable for quality control of Xinxue granules. PMID- 15807121 TI - [Characterization of Hg-binding metallothioneins in tissues of rat after oral intake of mercury chloride by size-exclusion chromatography coupled with inductively coupled plasma-mass spectrometry]. AB - Hg concentrations, total metallothioneins (MTs) and metal-binding MTs in tissues of the rat after oral intake of HgCl2 were determined by the inductively coupled plasma-mass spectrometry (ICP-MS), a modifield mecurry saturarion assay and the size-exclusion chormatography (SEC) coupled with ICP-MS, respectively. The results indicated that Hg accumulation in all of the rat tissues, including kidney, liver, testicle, brain, heart, serum and blood plasma, in HgCl2 group significantly increased compared with that in the control group, especiallyin kidney,liver and testicle. A large amount of MTs was inducted in these exposed rat tissues, especially in their kidneys and livers. The metal species binding with MTs obtained with SECICP-MS showed that the fractions of Hg-MTs in both kidney and liver of exposed rat were significantly higher compared with those of the control. In addition, higher amounts of Cu-MTs observed in exposed rat liver and kidney demomstrated the effect of MTs in the homeostatis of Cu and in the detoxification of Hg. PMID- 15807122 TI - [Determination of anions, cations and organic acids in the oil field water by ion chromatography]. AB - A method for the determination of anions, cations and organic acids in the oil field water with high chloride and sodium ions was developed by ion chromatography (IC) with a conductivity detector. The separation of the cations was achieved on an IonPac CS12A column with H2SO4 as the eluent. The separation of the anions and organic acids was achieved on an IonPac AS11-HC column with KOH as the eluent, and the detection was performed by a conductivity detection mode. The calibration graphs of peak area for all the analytes were linear over a wide range. The recoveries of the added standards were 90% - 110% . The relative standard derivations (RSDs) of the peak area of analytes were less than 8.0%. The detection limits for analytes ranged from 1.55 microg/L to 31.8 microg/L. PMID- 15807123 TI - [Determination of residual epichlorohydrin and sym-dichloroisopropyl alcohol in cationic etherified reagent by gas chromatography]. AB - A method for the determination of the residual epichlorohydrin and sym dichloroisopropyl alcohol in cationic etherified reagent by gas chromatography has been established. Methyl benzoate, which has high extraction efficiency for the two components, was used as extractant. With an HP-5 capillary column, the two components were baseline separated and they eluted before the extractant. The linear ranges achieved were 5 - 590 mg/kg for epichlorohydrin and 21 - 480 mg/kg for sym-dichloroisopropyl alcohol. The limits of detection were 1.2 mg/kg for epichlorohydrin and 2.2 mg/kg for sym-dichloroisopropyl alcohol. Recoveries for epichlorohydrin were 95.93% - 103.42% with relative standard deviations of 2.4% - 10.6% and those for sym-dichloroisopropyl alcohol were 98.54% - 107.40% with relative standard deviations of 6.6% -11.1%. The method is simple, fast, and convenient. PMID- 15807124 TI - [Analysis of low-polarity constituents of Reniochalina sp. by gas chromatography mass spectrometry]. AB - The sponge of Reniochalina sp. was collected from China Yellow Sea. The specimen (dry weight 100 g) was repeatedly extracted with cyclohexane. The combined crude extract was then evaporated in vacuo, and the dark red residue (5 g) was subjected to a silica gel column for chromatographic separation using gradient mixtures of ethyl acetate and cyclohexane as eluents. The fraction eluted with ethyl acetate-cyclohexane (1:9, v/v) was repeatedly separated by preparative liquid chromatography (HiQ sil C18-10 column, 250 mm x 21.2 mm i.d. ) to afford sample B and sample C as dark red oil. These two low-polarity samples were analyzed by gas chromatography-mass spectrometry (GC-MS) equipped with an HP-5 column (30 m x 0.25 mm i.d.). The oven temperature was raised from 40 degrees C to 250 degrees C at 10 degrees C /min and kept at 250 degrees C for 70 min. Using GC-MS coupled with 1H-nuclear magnetic resonance (1H-NMR), 18 compounds including alkanes, alkenes, fatty acids, fatty acid esters, phthalic acid and phthalic esters, were identified. The results provide to be helpful for in taxonomy of Reniochalina. PMID- 15807125 TI - [Monitoring of the residue of fosthiazate in water samples using solid-phase extraction coupled with gas chromatography/mass spectrometry]. AB - Solid-phase extraction (SPE) coupled with gas chromatography/mass spectrometry (GC/MS) was used to determine the fosthiazate residue in water samples. The water samples were first filtered through cellulose filters (0.45 microm pore size). A 100 mL volume of filtered water, in which 1 mL of methanol has been added, was then passed through a pre-conditioned 3 cm C18 cartridge at a flow-rate of 1.5 mL/min. Elution was performed by 1 mL of methanol. The eluant was finally dried under reduced pressure for solvent evaporation. The volume was quantitatively adjusted to 0.5 mL with methanol. The analysis was carried out on GC/MS. The mass spectrometer was operated in selected ion monitoring (SIM) mode. According to mass spectrum of fosthiazate, three selected ions at m/z of 126, 195, 283, respectively, were monitored for identification and quantification. High sensitivity and selectivity were achieved by using this method. The limit of detection for fosthiazate in water samples was determined to be 56.4 ng/L. The linearity was demonstrated over a wide range of concentrations covering from 0.282 to 141 microg/L. The recoveries were more than 85.5% and the relative standard deviations for the overall procedure were less than 4.42%. The fosthiazate residue was detected in the water samples from a pool near cropland where fosthiazate was used. The results demonstrate the suitability of the SPE GC/MS approach for the analysis of fosthiazate in water. PMID- 15807126 TI - [Study on determination of aflatoxins B1, B2, G1 and G2 in peanuts using OASIS HLB cartridges with post-column derivatization HPLC and fluorescence detection]. PMID- 15807127 TI - [Simultaneous determination of chloroxylenol, trilocarban and triclosan in cosmetics or household products by reversed-phase high performance liquid chromatography]. PMID- 15807128 TI - [Simultaneous determination of magnesium L-ascorbyl-2-phosphate, arbutin and kojic acid in cosmetics by reversed-phase high performance liquid chromatography]. PMID- 15807129 TI - [Determination of antioxidant in jet fuels by reversed-phase high performance liquid chromatography/mass spectrometry]. PMID- 15807130 TI - [Determination of EPA and DHA in marine microalgaes by GC]. PMID- 15807131 TI - [Determination of residual acetone in soybean phospholipids by headspace solid phase microextraction and gas chromatography]. PMID- 15807132 TI - [Determination of alpha-hydroxytrizolam and alpha hydroxyaplrazolam from urine by acetylation derivatization using gas chromatography electron capture detection]. PMID- 15807133 TI - [Determination of aroma components from apricot fruit by gas chromatography-mass spectrometry]. PMID- 15807134 TI - [Application of GC-FTIR to the analysis of high boiling residues in distillation tower bottom of ethyl benzene synthesized from dry gas]. PMID- 15807135 TI - [Determination of two ethylene glycol ether acetates in waterborne dopes with capillary gas chromatography]. PMID- 15807136 TI - Reality vs. Perception. PMID- 15807138 TI - Dentists, golf, and ethics. PMID- 15807137 TI - Thinking globally: dentists step up to help. PMID- 15807139 TI - And they'll take care of you ... PMID- 15807140 TI - The clinical significance and management of microleakage. Part one. AB - This research reviews relevant articles related to microleakage from 1966 to the present. The review discusses what the sensation of microleakage is clinically; how dentin permeability is related to microleakage; how the quality of dentin affects permeability and microleakage; what the effects of bacterial infiltration from microleakage can be, as well as the body's defenses against bacterial infiltration within the tooth complex; how caries and microleakage are related; and how various materials and procedures can be used to limit microleakage. PMID- 15807141 TI - Community Emergency Response Training. AB - My assessment of my Community Emergency Response Training experience is this: the training was excellent, and the experience was extremely valuable. That said, I would add that the one weakness that could be addressed in the program's design is the role of CERT teams in a biological attack by an infectious agent. PMID- 15807142 TI - Estate planning and charitable giving. PMID- 15807143 TI - Charitable giving and the Minnesota Dental Foundation. PMID- 15807144 TI - What's a dentist to do? Values, part two. PMID- 15807145 TI - History speaks to those who listen. PMID- 15807146 TI - Part two. Dentists' ideas for solutions: a 2004 survey of Greater Minnesota dentists. PMID- 15807147 TI - On group sequential procedures under variance heterogeneity. AB - In this paper, we consider group sequential procedures for clinical trials under variance heterogeneity. Group sequential procedures typically involve small samples at each interim analysis. We advocate Welch's correction for variance heterogeneity, and present a natural application of the significance level method for such situations. Currently available procedures are based on a large sample method, with no allowance for corrections of heterogeneity. Unless the sample size is large, the results are not valid. On the basis of simulation studies, comparing their abilities to control Type I error rates, we recommend using Welch's correction for sequential trials involving small samples under variance heterogeneity. PMID- 15807148 TI - Nonparametric methods for analysing the accuracy of diagnostic tests with multiple readers. AB - The evaluation of diagnostic agents or imaging procedures is governed by the same scientific and regulatory rules as that of other medical products. Receiver operating characteristic (ROC) curves, and especially the area under these ROC curves, are indices for the accuracy of a diagnostic test for continuous as well as ordinal data. The methodology of multivariate rank statistics for the nonparametric Behrens-Fisher problem is used to evaluate the accuracy of a diagnostic test in a complex factorial design with repeated measurements. Hypotheses are formulated by means of relative treatment effects and are tested by a multivariate extension of the Mann-Whitney statistic in a heteroscedastic model. The application of this method is demonstrated by the analysis of a data set from a diagnostic clinical trial. PMID- 15807149 TI - Revisiting proportion estimators. AB - Proportion estimators are quite frequently used in many application areas. The conventional proportion estimator (number of events divided by sample size) encounters a number of problems when the data are sparse as will be demonstrated in various settings. The problem of estimating its variance when sample sizes become small is rarely addressed in a satisfying framework. Specifically, we have in mind applications like the weighted risk difference in multicenter trials or stratifying risk ratio estimators (to adjust for potential confounders) in epidemiological studies. It is suggested to estimate p using the parametric family p(c) and p(1 - p) using p(c)(1 - p(c)), where p(c) = (X + c)/(n + 2c). We investigate the estimation problem of choosing c > or = 0 from various perspectives including minimizing the average mean squared error of p(c), average bias and average mean squared error of p(c)(1 - p(c)). The optimal value of c for minimizing the average mean squared error of p(c) is found to be independent of n and equals c = 1. The optimal value of c for minimizing the average mean squared error of p(c)(1 - p(c)) is found to be dependent of n with limiting value c = 0.833. This might justify to use a near-optimal value of c = 1 in practice which also turns out to be beneficial when constructing confidence intervals of the form p(c)+/-1.96 square root of np(c)(1 - p(c))/(n + 2c). PMID- 15807150 TI - Statistical methods for HIV dynamic studies in AIDS clinical trials. AB - Studies of HIV dynamics in AIDS research are very important for understanding pathogenesis of HIV infection and for assessing the potency of antiviral therapies. Since the viral dynamic results from clinical data were first published by Ho et al. and Wei et al., the study of HIV-1 dynamics in vivo has drawn a great attention from AIDS clinicians and researchers. Although the important findings from HIV dynamic studies have been published in many prestigious scientific journals, statistical methods for estimating viral dynamic parameters have not been paid enough attention by HIV dynamic investigators. The estimation methods in many viral dynamic studies are very crude and inefficient. In this paper, we review the statistical methods and mathematical models for HIV dynamic data analysis developed in recent years. We also address some practical issues and share our experiences in the design and analysis of viral dynamic studies. Some principles and guidelines for the design and analysis of viral dynamic studies are provided. The methodologies reviewed in this paper are also applicable to studies of other viruses such as hepatitis B virus or hepatitis C virus. We also pose some challenging statistical problems in this area in order to stimulate further study by the statistical research community. PMID- 15807151 TI - [Global strategies in prevention of infectious diseases on the turn of the second and third millennium: expectation versus reality]. AB - Prevention has been the greatest success in the field of infectious diseases. Control of infectious diseases is being challenged by many factors, like standards of living, human behaviour, mass population movements, emergence of infectious like HIV/AIDS and SARS, the re-emergence of infections such as tuberculosis, development of resistance in bacteria, viruses and parasites, modern rate of global travel, etc. Although great advances have been made, a considerable burden of morbidity and mortality from infectious diseases remains globally. The aim of any control program can be defined by three levels- containment, elimination and eradication. Prevention methods are many aspects, including introduction of prevention guidelines, low cost interventions, vector control, surveillance, and vaccination, etc. PMID- 15807152 TI - [Herd immunity and effectiveness of vaccination]. AB - The effectiveness of vaccinations is discussed in relation to vaccine efficacy and effectiveness of vaccination programs. The types of epidemiological studies used in the assessment of vaccine effectiveness are presented, and most common sources of bias in such studies are listed. Basic formulas for calculation of vaccine effectiveness are given as applied for cohort and case-control studies. The definitions and ways of estimation of indicators of herd immunity as applied to the analysis of the effectiveness of vaccinations are presented. PMID- 15807153 TI - [CMV infections]. AB - Infections with cytomegalovirus are widespread among populations. The frequency of anti-CMV bodies occurrence in healthy adults is from 40% in Europe to 100% in the Third World countries. The research carried out in Poland on two thousand healthy women at productive age showed in more than half of them the presence of anti-CMV bodies before pregnancy. It is especially significant in the context of the possibility of primary CMV infection in gravidas and the transmission into the fetus. Confirmation of the primary CMV infection in women at an early stage of pregnancy requires further prenatal diagnosis. CMV is asymptomatic in 99% of cases. It is especially dangerous for neonates, in whom heavy cytomegalovirus disease is connected with immaturity of the immunological system, and for people with immunological system disorders (after transplantations or HIV infections). Clinical forms of CMV infections have been presented here, with some of them being illustrated with descriptions of cases observed at the Clinic; methods of CMV infection treatment in various groups of patients, as well as the state of knowledge about anti-CMV vaccine are also presented. PMID- 15807154 TI - [Smallpox--historical or real threat]. AB - Presently, there is no real possibility of natural re-emergence of smallpox virus, which was eradicated globally more then 25 years ago. During the last decade the possibility of use of smallpox virus as a biological weapon by a criminal organisation was emphasised. The re-emergence of smallpox virus would lead to unprecedented disaster. Theoretical models indicated that only extremely strict and enforced interventions could stop the spread of epidemic, but the assumptions of these models were unrealistic. Presently, there are limited stocks of the first generation smallpox vaccine left in the world. This vaccine, as well as the second-generation vaccine are associated with multiple adverse events, including fatalities and may not be accepted by society. Much safer vaccines are now being developed. Strategic plan of prophylactic vaccinations requires defining the groups to be immunised in the first place and whether immunisation should start before or after a first smallpox case would occur. PMID- 15807155 TI - [Risk of influenza in healthy children--vaccination benefits]. PMID- 15807156 TI - [Prevention of pertussis and high expectations concerning vaccines]. AB - The basic vaccine used in the prevention of pertussis is the combined vaccine including a whole cell pertussis component and tetanus and diphtheria toxoids. Although this type of vaccine has been used more than 50 years in USA and more than 40 years in Poland it is still effective what can be evidenced by the decreased number of pertussis cases since the vaccine has been implemented. There are however some evidences that the whole cell vaccine may lead to the acute encephalopathy, fever seizures, hypotonic-hyporeactive episodes, inconsolable crying or anaphylactic reactions. But still is a lack of convincing evidences that the vaccine may be a cause of persistent brain damage. It was also shown that the longer is the period after the last dose of the vaccine the lower effectiveness was observed. Improving the safety of the pertussis vaccine was the reason of introducing the acellular vaccines in the eightieth. All these products contain pertussis toxoid and some of them contain also filamentous hemagglutinin, pertactin and fimbrial agglutinogens. Some published studies have shown that the effectiveness of these vaccines is similar to the whole cells vaccines and that the incidence of some adverse events especially seizures, hypotonic-hyporeactive episodes and inconsolable crying is lower. PMID- 15807157 TI - [Combined vaccines in European Union]. AB - Combined vaccines are urgently needed because of the increasing number of recommended vaccines for children. Combined vaccines could be considered as having the same priming effect as monovalent vaccines. They demonstrated a good reactogenicity and tolerability profile. Combined vaccines are more comfortable option for the infant vaccination providing immunity against four or more important childhood diseases with a single injection at each visit. PMID- 15807158 TI - [Epidemiologic aspects of viral hepatitis A and vaccination]. AB - Epidemiological problems connected with hepatitis A in twentieth century and in the first years of twenty-first were mentioned with special attention of self limiting of cases in the periods of very high, high, medium, low and very low endemicity. Decreasing of percentage of immunised persons was observed especially among children and young persons. It may be a cause in future of increasing of numbers of cases of hepatitis A even epidemics. Therefore vaccination against hepatitis A some people in risk groups is now recommended. Especially are included persons with some professions (health workers, persons employed in food and meals industry, travellers to the region of high or medium endemicity etc), persons living in territory of poor sanitary conditions, children, adolescents and young adults as well as persons with some chronic diseases, drug addicts, homosexuals. PMID- 15807159 TI - [Analysis of HBV source in patients hospitalized at the Department of Hematology and Clinical Immunology of the Medical Academy in Warsaw in the years 2000-2003]. AB - The purpose of the study was to evaluate routes of HBV transmission in 140 patients hospitalized in the Department of Hepatology and AIDS, Medical University of Warsaw in the years 2000--2003. Authors found two major routes of HBV infection in the observed patients: drug abuse and the unknown route in the group of young people and infection transmitted at the non-surgical wards among the patients older than 40 years. PMID- 15807160 TI - [Combined vaccine against hepatitis A and B]. AB - Among the infectious virus hepatitis diseases, types A and B can be successfully prevented with vaccinations. WHO recommend effective control of hepatitis A through immunisation, especially among populations with indirect or high endemicity. Since 1991 WHO has been recommending vaccinations against hepatitis B. It is planned to introduce it in all countries by the year 2007. Vaccinations against hepatitis A & B are recommended for people travelling to the regions endemic for these infections or for those with high professional risk of HAV and HBV. Presented here are the characteristic features of the vaccine, the analysis concerning the evaluation of its effectiveness, comparison of this effectiveness with monovalent vaccine effectiveness, as well as evaluation of the tolerance and safety. PMID- 15807161 TI - [HIV and HCV infections--clinical implications]. AB - Coexisting HIV and HCV infection involves approx. 10 million people in the world. It is most common in drug addicts taking drug injections as well as in haemophiliacs. In the research we have confronted the existing views on natural history progression in hepatitis C in HIV infected, with our own study. The existing recommendations for hepatitis C treatment have also been presented. It was pointed out that there is a possibility of interaction between the drugs applied in the treatment and the antiretrovirus therapy and what is connected with it, the possibility of potentially adverse actions. It was also shown that early treatment of hepatitis C in HIV infected slows down the progress of liver fibrosis, prevents liver failure, improves HA-Art tolerance, increases immunological reconstruction and improves the quality of the patients' lives. PMID- 15807162 TI - [Chance of hepatic fibrosis treatment in chronic hepatitis C]. AB - Treatment of liver fibrosis in chronic hepatitis C should affect at least one among elements of fibrosis pathogenesis. At the present time the only accessible therapeutic method is clearance of HCV infection using combined treatment with pegylated interferon alpha and ribavirin. Other therapeutic opportunities, that can be applied shortly include: reduction of hepatocytes apoptosis by caspase inhibitors, decrease of inflammation intensity using interleukin-10, and inhibition of hepatic stellate cells transformation into myofibroblasts with interferon-gamma. Efficiency of purified phospholipids acting as metalloproteinases inducers or antagonists of profibrogenic cytokines need further evaluation. PMID- 15807163 TI - [Vaccination programs in Poland]. PMID- 15807164 TI - [Vaccination priorities for traveller's children]. AB - Routine, recommended and required immunization are important for traveller's children. All required immunization. The most frequent vaccine vaccine preventable infection travellers to developing counties in hepatitis A. Influenza is estimated to be 1%. Routine immunization against poliomyelitis is important for travel to north Africa and parts of Asia. It should be considered that many immunizations are recommended for special risk groups only. If health professionals do not comply with rule, serious problems my arise as a traveller my be refused entry to the country. PMID- 15807165 TI - [Vaccination for HIV positive travelers]. AB - People with HIV+ travelling around the world comprise a unique group regarding immunological disorders which can be intensified due to, e.g. climatic and time zones. Increased exposure of vulnerable tourists to various biological factors (viruses, bacteria, fungi, and parasites) is hazardous to their health. Travellers are advised to active prophylaxis as far as endemic diseases of particular globe regions are concerned. Limitations in vaccine use in HIV+ travellers are due to the kind of biological material contained in preparations. Live vaccines in those with impaired immunity are limited as it is widely known. It is absolutely contraindicated to vaccinate patients with AIDS against tuberculosis, typhoid fever, and yellow fever. Killed vaccines are considered safe and recombined ones are recommended. Vaccines against HBV, HAV, Haemophilus influenzae b (Hib), influenza, and pneumococcus are indicated. However, the kind of prophylaxis should be individualized, decided by a managing physician in concord with an appropriate referential center. PMID- 15807166 TI - [Yellow fever]. AB - Yellow fever is a mosquito born viral illness that causes hemorrhagic fever in tropical Africa and South America. An estimated 200 000 people are still infected annually. There is no antiviral therapy available and vaccination is major strategy in the control yellow fever. From 60 years ago is available very safe and efficient vaccine. The vaccine is a live attenuated virus strain 17D. Recently identified vaccine associated viscerotropic diseases (YFV-AVD). This paper describes knowledge about yellow fever disease, vaccine associated adverse events and actually recommendation for vaccine use. PMID- 15807167 TI - [Varicella--mild or dangerous disease]. AB - OBJECTIVE: The aim was an analysis of causes of varicella complications in children hospitalized in the Department if Infectious Diseases In Childhood of Medical University in Warsaw. METHODS AND MAIN OBSERVATIONS: 105 children from 15 day to 15 years of age was observed. 58.1% were male. The median age was 5,8 years. RESULTS AND CONCLUSIONS: Bacterial skin infections and neurological complications were the most frequent complications. Sepsis was diagnosed in 3 children. Vaccination program against varicella can reduce the risk of very severe complications. PMID- 15807168 TI - [Alternative immunization schedule and combined vaccines in paediatric practice]. AB - European countries individually implement their national immunisation schedules adjusted to their economic situation, epidemiological risks and other socio political factors. The polish schedule of compulsory vaccination includes an obligatory part financed by the state and the recommended one financed individually by parents of patients themselves. The officially registered multi component vaccines my become an alternative for the obligatory schedules. Benefits of combined vaccine comprise: reduced number injections per visit and reduced number of exposures to possible injection pain; less or no exposure to thiomersal; less time spent in doctor visit, less waste and increased injection safety with fewer syringes, reduced immunisation programme costs and facilitated addition data collection through easier documentation and simplified implementation of immunisation programmes. PMID- 15807169 TI - [Vaccinations as a cause of children hospitalisation in Neuroinfection Department of the John Paul II Hospital in Cracow between 2002--2004]. AB - A retrospective study was conducted among 205 children chosen from a group of 2728 children hospitalised in Neuroinfection Department of the John Paul 2nd Hospital In Cracow between 1/1/2002 and 6/30/2004. The data was taken from medical documentation from the Neuroinfection Department and Province Dispensary of Inoculation in the Hospital. Patients was divided into two groups: first one contained children from group of increased risk, patients hospitalised electively in the Dispensary to be under observation in hospital environment on day of execution of inoculation; and second one contained children sent to the Neuroinfection Department from other institutions in consequence of showed symptoms of adverse event following immunization (AEFI) Within both groups' causes of hospitalisation was determined and given types of vaccinations was qualified as the most often cause of hospitalisation. It appeared that predominantly cause of hospitalisation was misgivings about course of the inoculation process for children with neurological diseases, which had been inoculated DTaP according to plan. However the most often to cause AEFI was vaccination DTP with full-cell pertussis component. PMID- 15807170 TI - [Diagnostic methods of postvaccinal complications after administration of BCG vaccine]. PMID- 15807171 TI - [Effectiveness of vaccination against hepatitis B in HIV infected children]. AB - OBJECTIVE: The aim of the study was to evaluate hepatitis B vaccination effectiveness in HIV infected children. METHODS: 45 children vertically infected with HIV who received hepatitis B vaccination were evaluated. Anti-HBs antibodies were assayed to established vaccination efficiency and were repeated every 6-12 months. No-responders received the next vaccination schedule with doubled dose. Children with antibody levels < 100 IU/ml were boostered. All the children have been receiving HAART. MAIN OBSERVATIONS AND RESULTS: 35/45 (77,8%) children had anti-HBs antibodies > 100 IU/ml, including 20 with anti-HBs antibodies > or = 1000 IU/ml (32 children without immunodeficiency, 2 with moderate and 1 with severe immunodeficiency). Anti-HBs level 8-100 IU/ml was observed in 6/46 (13,3%) children (5 children without immunodeficiency and 1 with severe immunodeficiency). 4/46 (8.9%) children had no anti-HBs antibodies (2 children without immunodeficiency, 1 with moderate and 1 with severe immunodeficiency). CONCLUSIONS: In HIV infected children anti-HBs antibodies should be assayed to establish hepatitis B vaccination efficiency and repeated every 6-12 months. PMID- 15807172 TI - [Complications of chickenpox as reason for children's hospitalization]. AB - Chickenpox is self limiting disease, with potentially dangerous course. Chickenpox complications can evoke the necessity of hospitalization. Assess the types and courses of chickenpox complications in child patients hospitalized in Provincial Hospital of Infectious Diseases in Bydogoszcz between 1999 and 2003. Cases of chickenpox complications being the reason of children hospitalization were retrospectively analyzed. The total number of children hospitalized because of chickenpox complication is 153. Patients age ranged from 18 days to 18 years, with average of 5,4 years. 62% of children were younger than 5 and, 1/3 children were younger than 1. The average period of hospitalization was 7 days. 17% of patients stayed in hospital longer than 10 days. 53% of patients were male. In 5 cases chronic diseases were diagnosed. The most common reason of hospitalization connected with chickenpox were symptoms of alimentary canal disorder (30%), respiratory tract inflammations, with pneumonia and bronchitis in the lead (30/47). Neurological complications during chickenpox occurred in 23 of 153 hospitalized (15%): febrile convulsions - 6/153, cerebellar ataxia - 9/153, meningitis and brain fever - 6/153, peripheral nerve - 2/153. Bacterial skin infection as the reason of hospitalization of 16 children, hepatitis of 3 and joints inflammation of 1 child. PMID- 15807174 TI - [Treatment of chronic hepatitis B]. AB - The aim of the study was comparison of the efficacy of treatment with INF and lamivudine in chronic hepatitis B. 103 patients with chronic hepatitis B were included in this trial and divided into two groups. Group I consisted of 34 patients treated with INF-alpha (5MU 3x per week during 4-6 months), group II included 69 patients treated with lamivudine (100mg per day during 1-4 years). Virologic, serologic and biochemical factors at ETR were analyzed. Costs of these two therapies were estimated too. In group I HBV DNA clearance was observed in 26/34 cases (76%), seroconversion HBeAg/anti-HBe in 5/34 (15%) and normalization of ALT in 22/30 patients (73%). In group II virological response occurred in 37/69 cases (53%), serological in 16/41 (39%) and biochemical in 34/50 (68%). Patients from group I suffered from adverse events typical for INF therapy. No adverse events were observed in group II. Cost of virological response in group I was 4 500 zl, in group II 8 000 zl, biochemical--INF 4 700 zl, lamivudine 5 700 zl and serological--group I 18 600 zl, in group II 18 700 zl. PMID- 15807175 TI - Lightning injuries and acute renal failure: a review. AB - Lightning strikes cause multimodal injuries in victims, and although the number of deaths due to lightning is reportedly in the area of 1,000 deaths per year, they cause significant morbidity in many others. A major complication of a lightning strike is acute renal failure (ARF). The true incidence of ARF due to lightning injuries worldwide is difficult to ascertain because of significant underreporting, due in large measure to cultural and sociodemographic factors. Its incidence is reportedly rare in some literature and significant in others. However, lightning's potential to cause ARF due to myoglobinuria has been noted by many authors. Prompt treatment of myoglobinuric patients prevents the development of ARF. ARF due to direct organ damage by lightning is virtually unheard of. In this article, the current mechanisms of lightning injuries leading to muscle damage, myoglobinuria, and subsequent ARF are discussed, as well as signs and symptoms, laboratory investigations, and patient management. PMID- 15807176 TI - Influence of parenteral iron therapy and oral vitamin E supplementation on neutrophil respiratory burst in chronic hemodialysis patients. AB - BACKGROUND: Bioincompatibility of hemodialysis (HD) membranes is responsible for neutrophil activation leading to oxidative stress, which can be further increased by intravenous (IV) iron (Fe) administration. The aim of our study was to monitor neutrophil respiratory burst during HD and to find out whether this process is influenced by IV Fe and oral vitamin E administration. METHODS: Within four HD sessions, blood samples were taken from seven chronic HD patients at time 0 (before HD), 60, 70, and 130 min of HD session. Neutrophil respiratory burst was assessed by luminol-enhanced chemiluminescence (CL). Plasma advanced oxidation protein products (AOPP) concentration was measured spectrophotometrically. During the second and the fourth HD, 62.5 mg of sodium ferric gluconate was applied IV in the 65th minute of HD. Before the last two HD, the patients were given orally 200 mg of vitamin E daily for 7 days. Patient's results were compared with healthy controls. RESULTS: Predialysis CL is higher in patients than in controls (1,926 +/- 436 vs. 1,083 +/- 325 RLU, p<.01). CL decreases in the 60th min of HD (1,926 +/- 436 vs. 1,220 +/- 599 RLU, p<.05); thereafter, it remains stable. After Fe application, CL increases at time 130 compared with CL at time 60 (1,303 +/- 269 vs. 877 +/- 292 RLU, p<.05). AOPP concentration is higher in patients than in controls (137.5 +/- 42.7 vs. 88.9 +/- 24.8 micromol/L, p<.01) and remains unaffected by vitamin E supplementation. After vitamin E intake, predialysis CL remains significantly higher than in controls, and changes in CL during HD are minimal despite Fe administration. CONCLUSION: HD patients' neutrophils generate more oxygen radicals than in healthy individuals. This production decreases during HD and then increases after IV Fe administration. Short-term vitamin E administration attenuates this fluctuation of neutrophil oxidative metabolism, without affecting the total degree of oxidative stress. PMID- 15807177 TI - Metabolic acidosis aggravation and hyperkaliemia in hemodialysis patients treated by sevelamer hydrochloride. AB - Reports on acid-base side effects of sevelamer hydrochloride (SH), a new aluminum (Al)- and calcium (Ca)-free phosphate binder are rare and conflicting. In a retrospective analysis, we evaluated SH impact on metabolic acidosis and serum potassium (K) in hemodialysis (HD) patients. Two groups of stable HD patients were studied. Group A included 17 patients, M/F=15/2, 64 (42-80) years old, dialyzed since 130 (34-253) months, under SH for 24 months. Group B serving as controls was made of 7 patients, M/F=4/3, 67 (48-91) years old, dialyzed since 67 (27-174) months, under CaCO3 and/or Al(OH)3 as phosphate binders also for 24 months. Bicarbonate (BIC), K, Ca, phosphorus (P), Ca x P, alkaline phosphatase (ALP), and intact parathyroid hormone (iPTH) were recorded before (MO) and at the end (M24) of 24-month SH or CaCO3-Al(OH)3 treatment in group A and B patients. In group A, BIC fell from 20.02 +/- 1.43 to 17.89 +/- 2.30 mEq/ L, P=.002; and K rose from 5.45 +/- 0.51 to 5.75 +/- 0.49 mEq/L, P=0.02. In group B, BIC (19.8 +/- 3.03 to 19.0 +/- 3.3 mEq/L) and K (5.01 +/- 0.8 to 4.9 +/- 1.1 mEq/L) had nonsignificant changes. In group A, iPTH rose from 132.82 +/- 124.08 to 326.89 +/ 283.91 pg/mL, P=.0008; P fell from 5.92 +/- 1.48 to 4.9 +/- 1.01, P=.02; and Ca x P decreased from 52.04 +/- 9.7 to 45.58 +/- 10.42 mg2/dL2, P=.04. In group B, changes in iPTH from 240.71 +/- 174.7 to 318.57 +/- 260.2 pg/mL, P from 4.9 +/- 0.5 to 4.8 +/- 1.3 mg/dL, and CaxP product from 44.3 +/- 6.6 to 44 +/- 11.2 mg2/dL2 were nonsignificant. The changes observed in Ca and ALP in both groups were nonsignificant. Correlations in group A between metabolic acidosis (BIC) and SH doses, or iPTH and BIC, Ca, or P changes, were also found to be nonsignificant. Long-term use of SH, effectively controlling serum P levels and Ca x P values, is associated with acidosis aggravation and hyperkaliemia. Worsening of secondary hyperparathyroidism, also noted, needs to be confirmed and could be related to Ca/Al salt discontinuation and to metabolic acidosis aggravation itself. PMID- 15807178 TI - Clinical comparisons of 1,498 chronic renal failure patients with and without tuberculosis. AB - OBJECTIVE: To investigate the clinical characteristics of chronic renal failure (CRF) patients suffering from tuberculosis. METHODS: Clinical materials from CRF inpatients in our department suffering from tuberculosis from 1997 to 2003 were investigated and compared with similar data from tuberculosis patients without CRF in Chongqing City (China) during the same period. RESULTS: Of the 1,498 tested CRF inpatients, 71 suffered from tuberculosis, more than half of which suffered from atypical type V tuberculosis (lesion spot uncertain). The incidence of tuberculosis in the CRF inpatients (4740/100,000) was significantly higher than that in Chongqing City (151/100,000; P<.001). We found an inverse association between renal function and tuberculosis; the worse the renal function, the higher the tuberculosis incidence. CRF patients suffering from tuberculosis had fewer positive antituberculosis-antibody and purified protein derivative tests (12.7% and 15.5%, respectively) than did tuberculosis patients without CRF (72.1% and 58.1%, respectively; P<.05). In CRF patients with tuberculosis, the two-thirds dosage antituberculosis drug treatment was effective. Twelve cases had good outcome after a 1-year course of antituberculosis treatment and then kidney transplantation. In contrast, 4 patients who received less than half a year of antituberculosis treatment died after kidney transplantation, in association with relapsed tuberculosis. CONCLUSION: CRF patients are susceptible to tuberculosis, especially to atypical tuberculosis outside the lung. Thus, antituberculosis treatment might be necessary in CRF patients found to be suffering from chronic fevers of unknown origin. Furthermore, it appears to be safe for patients with CRF plus tuberculosis to receive kidney transplantation after 1 year of antituberculosis treatment. PMID- 15807179 TI - Estrogen receptor (ER) gene polymorphism may predict the bone mineral density response to raloxifene in postmenopausal women on chronic hemodialysis. AB - The estrogen receptor (ER) gene has been considered as a candidate genetic marker for osteoporosis, and PvuII and XbaI polymorphisms of the ERalpha gene have been associated with low bone mineral density (BMD). We investigated whether ER polymorphism could predict the response of BMD in 28 postmenopausal women on hemodialysis with marked osteopenia or osteoporosis, randomized to receive raloxifene, a selective estrogen receptor modulator (SERM), or placebo for 1 year. BMD was assessed by dual X-ray absorptiometry and PvuII and XbaI restriction fragment-length polymorphism of the ER gene was determined using polymerase chain reaction. Baseline lumbar spine or femoral neck BMD parameters were not different between patients presenting either homozygous PP or xx when compared with heterozygous Pp or Xx genotypes. After 1 year, patients on raloxifene, presenting with PP or xx genotypes (but not those with Pp or Xx), showed a significantly higher mean lumbar spine BMD (0.942 +/- 0.18 vs. 0.925 +/- 0.17 g/cm2, p < .01) and lower serum pyridinoline (19.7 +/- 9.7 vs. 30.6 +/- 16.5 nmol/L, p < .02) when compared with baseline values. No changes were detected in the placebo-treated patients or in the femur neck sites. In conclusion, after 1 year on raloxifene, postmenopausal osteoporotic women on chronic hemodialysis, homozygous for the P or x (PP or xx) alleles of the ER, exhibited a better lumbar spine BMD response and decreased serum pyridinoline values when compared with heterozygous women (Pp or Xx), suggesting that ERalpha allelic variants may explain, at least in part, the different outcomes after treatment of osteoporosis with SERM. PMID- 15807180 TI - Parathyroid hormone levels, calcium-channel blockers, and the dyslipidemia of nondiabetic hemodialysis patients. AB - BACKGROUND: Experimental studies have shown that increased levels of parathyroid hormone (PTH) in uremia may cause elevation of intracellular calcium, predisposing to insulin resistance and lipid metabolism abnormalities. Administration of calcium-channel blockers (CCBs) in these models protects against the development of lipid profile abnormalities. This study evaluates the combined effect of intact PTH (iPTH) levels and administration of CCB on the lipid profiles of nondiabetic hemodialysis patients. METHODS: One hundred and eight non-diabetic hemodialysis patients were studied for 6 months. The population was divided into four groups, according to iPTH levels and administration of CCB: (A) iPTH<70 pg/mL, administration of CCB (n=16), (B) iPTH>300 pg/mL without administration of CCB (n=43), (C) iPTH<70 pg/mL without CCB administration (n=19), and (D) iPTH>300 pg/mL with CCB administration (n=30). Serum concentrations of total cholesterol, high-density lipoprotein (HDL), triglycerides, and albumin were measured on a monthly basis. RESULTS: All results are shown as mean SE. Total cholesterol values (in mg/ dL) were for group (A) 186 +/- 4, for group (B) 205 +/- 3, for group (C) 200 +/- 3, and for group (D) 203 +/ 4 [p NS between (C) and (D), p<.05 for all other comparisons]. Triglycerides values (in mg/dL) were for group (A) 171 +/- 9, for group (B) 199 +/- 6, for group (C) 190 +/- 6, and for group (D) 191 +/- 9 (p NS for all comparisons). HDL values (in mg/dL) were for group (A) 43.8 +/- 1, for group (B) 35.8 +/- 1, for group (C) 38.3 +/- 0.7, and for group (D) 37.2 +/- 0.7 mg/dL [p NS between (C) and (D), p<.001 for all other comparisons]. Low-density lipoprotein values (in mg/dL) were for group (A) 107.6 +/- 4.4, for group (B) 149.3 +/- 2.5, for group (C) 131.2 +/- 2.9, and for group (D) 126.8 +/- 4.1 [p NS between (C) and (D), p<.001 for all other comparisons]. Atherogenic index values, calculated as [triglycerides/HDL] ratio, were for group (A) 4.6 +/- 0.04 , for group (B) 6.2 +/ 0.04, for group (C) 4.9 +/- 0.03, and for group (D) 5.9 +/- 0.03 [p NS between (C) and (D), p<.004 for all other comparisons]. CONCLUSION: In nondiabetic hemodialysis patients, lipid profile abnormalities often accompany high levels of iPTH. The decrease in iPTH and/or the administration of CCB are accompanied by significant improvements in the main lipid profiles, including the atherogenic index. PMID- 15807181 TI - Silent myocardial ischemia and high-grade ventricular arrhythmias in patients on maintenance hemodialysis. AB - The occurrence of silent myocardial ischemia (SMI) and serious arrhythmias during hemodialysis (HD) has been well documented. However, it is unclear whether these changes are due to epicardial coronary artery disease (CAD). We conducted a prospective study to assess whether SMI and arrhythmias during HD correlated with angiographically demonstrable CAD. Twenty-three patients with end-stage renal disease on maintenance HD underwent 48-hour Holter monitoring, beginning 24 hours prior to a HD session. All patients underwent biochemical evaluation, coronary angiography, and echocardiography. Holter monitoring showed SMI during HD in 22% cases. A significant increase in the frequency of ventricular ectopics (VEs) was noted during and after HD. Patients who showed SMI during HD and VEs prior to initiation of dialysis were more likely to develop significant ventricular arrhythmias during and after HD. Epicardial CAD was documented in four patients, and it did not correlate with SMI. To conclude, HD is an arrhythmogenic process. SMI during dialysis is probably not due to epicardial CAD but predisposes to clinically significant ventricular arrhythmias during and after HD. The cause of SMI during HD in patients without demonstrable CAD needs further investigation. PMID- 15807182 TI - Effect of erythropoietin therapy on red cells filterability and left ventricular mass in predialysis patients. AB - BACKGROUND: Cardiovascular complications are the leading cause of mortality in patients with end-stage renal disease. Left ventricular hypertrophy (LVH) is recognized as an independent risk factor for cardiovascular morbidity and mortality. At the onset of dialysis, more than 70% of the patients with chronic kidney disease have echocardiographic evidence of LVH. Anemia, increased red cells filterability time (RCFT), and blood viscosity are known to induce LVH. AIM: To evaluate, prospectively, the effects of erythropoietin (EPO) therapy for 20 weeks on RCFT and left ventricular mass (LVM). PATIENTS AND METHODS: Twenty uremic and anemic predialysis patients with creatinine clearance test below 35 mL/min were studied. RCFT test and three-dimensional echocardiography were performed at 0, 10, and 20 weeks. RESULTS: EPO therapy for 20 weeks did not adversely affect renal function and did not significantly change the mean blood pressure. It significantly increased the hemoglobin and fibrinogen levels, and decreased RCFT and LVM (p < .01). CONCLUSION: Although correction of anemia can contribute to regression of LVM, we speculate that an increasing number of cells with normalized viscoelastic properties and a direct effect of EPO on erythrocytes and myocardiocytes, through specific receptors, may also play an important role. PMID- 15807183 TI - Effect of off-pump and on-pump coronary artery bypass grafting on renal function. AB - BACKGROUND: Coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) has the risk of renal dysfunction. The cause of renal dysfunction after CPB is multifactorial, such as nonpulsatile flow, renal hypoperfusion, hypothermia, and duration of CPB. This study compared off-pump technique with on-pump technique on renal function in patients who underwent CABG. METHODS: Sixty patients with normal preoperative renal functions undergoing CABG were randomly assigned to conventional revascularization with CPB (on-pump) or beating heart revascularization (off-pump). Renal functions were assessed up to 10 days postoperatively. RESULTS: Creatinine clearance was found to be significantly higher in the off-pump group than in the on-pump group (p<.05). The off-pump group had significantly less increase in creatinine levels when compared with the on-pump group (p<.05). The free water clearance values decreased similarly in both groups; however, the recovery was more prompt in the off-pump group (p<.05). No significant differences were found in the prevalence of postoperative hemodialysis. CONCLUSION: The off-pump technique may provide a positive contribution and sufficient protection on postoperative renal functions in patients undergoing CABG. PMID- 15807184 TI - Multicystic dysplastic kidney: conservative management and follow-up. AB - This study reports our experience in the conservative management of multicystic dysplastic kidney (MCDK) disease. Between 1992 and 2003, 35 children, 18 male (51.4%) and 17 female (48.6%), were diagnosed with MCDK and prospectively followed at the Riyadh Armed Forces Hospital. Static scintigraphy with dimercaptosuccinic acid and micturating cystourethrogram were performed in all patients. Ultrasonography (US) examinations were conducted every 3 months, and then annually. Mean age at diagnosis during the postnatal period was 2.4 days. Median follow-up time was 33.75 months. Follow-up US examinations showed partial involution in 17 (48.6%), complete involution in 13 (37.1%), and 5 (14.3%) showed an unchanged size without any pathological manifestations. The mean age at complete or partial involution of the lesion was 43.7 months. No child developed complications such as hypertension or tumors, and all maintained normal growth and development. In conclusion, MCDK lesions are often benign, and the affected kidneys tend to show involution with time. PMID- 15807185 TI - Effect of 1-year oral alpha-tocopherol administration on anticardiolipin antibodies in hemodialysis patients. AB - BACKGROUND: Anticardiolipin antibodies (ACA) have been related to an increased incidence of thrombotic episodes and atherosclerosis progression. ACA levels are elevated in hemodialysis (HD) patients. Atheroembolic episodes are the major cause of morbidity and mortality in this population. Oxidative stress has been implicated in ACA formation, and it is increased in HD patients. Vitamin E is a known antioxidant factor. In this study, the effects of prolonged oral alpha tocopherol administration on ACA levels were evaluated. METHODS: Serum anticardiolipin IgG antibodies (ACA-IgG) and IgM antibodies (ACA-IgM) levels were evaluated in 27 stable HD patients and 22 healthy volunteers. Then measurements were performed in the patients' group after oral administration of alpha tocopherol at a dose of 500 mg/d for a 1-year period. ACA levels were assessed by solid-phase enzyme immunoassay. RESULTS: ACA-IgG levels were higher in HD patients compared with control (13.3 +/- 6.64 GPL/mL vs. 7.727 +/- 18.305 GPL/mL, p < .001). This was not the case for ACA-IgM levels (2.96 +/- 4.18 MPL/mL vs. 1.386 +/- 2.636 MPL/mL, p=.17). alpha-Tocopherol administration resulted in a further increase in ACA-IgG (26.7 +/- 14.7 GPL/mL vs. 13.3 +/- 6.64 GPL/mL, p < .001) and ACA-IgM levels (8.17 +/- 1.95 MPL/mL vs. 2.96 +/- 4.18 MPL/mL, p < .001) in HD patients. CONCLUSIONS: Prolonged oral alpha-tocopherol administration in HD patients increases ACA levels. The mechanism and the clinical significance of this finding need further evaluation. PMID- 15807186 TI - Hemodynamic effects of anemia correction by recombinant human erythropoietin in predialysis patients with renal failure. AB - Most patients with chronic renal failure have anemia, which can be corrected by recombinant human erythropoietin (rHuEpo) treatment. Increase in arterial pressure (AP) was reported in some studies and was related to higher systemic vascular resistance induced either by the rise of erythrocyte mass or the change in various endogenous vasopressors, including the direct action of rHuEpo itself. We investigated the effect of rHuEpo treatment on hemodynamic variables, including small and large arterial compliance in 20 patients with chronic renal failure who were not receiving dialysis (CCT 29 +/- 12 mL/min), with Hb levels of 40.4 +/- 0.58 g/dL. They were treated with 2,000 units intravenously followed by 80 to 120 s/c units/kg/body weight, with dosage titration according to Hb level. Noninvasive hemodynamic evaluation was performed before the first rHuEpo treatment, 30 min after the first IV rHuEpo administration and at least 3 months later when target hemoglobin (Hb) and hematocrit (Hct) were reached. No rise in AP occurred after rHuEpo administration either short term or long term. The significant hemodynamic changes were a fall in pulse pressure and a rise in large artery compliance, with no change in small artery compliance after 3 months of rHuEpo treatment when Hb and Hct levels were corrected. These findings show improvement in arterial stiffness when Hb is corrected with rHuEpo treatment. PMID- 15807187 TI - Comparisons between oral pulse alfacalcidol therapy and daily therapy in maintenance hemodialysis patients with secondary hyperparathyroidism: a randomized, controlled, and multicenter study. AB - OBJECTIVE: To investigate the efficacy and safety of 1alpha-(OH)-D3 high-dose pulse therapy or daily low-dose therapy in secondary hyperthyroidism in maintenance hemodialysis patients in China. METHODS: Maintenance hemodialysis patients of both gender with intact parathyroid hormone (iPTH) level above 200 pg/mL were randomly divided into a pulse group and a daily group. They were treated for 20 weeks, with 2 microg oral Alfacalcidol twice weekly or thrice weekly in the pulse group, and 0.5 microg oral Alfacalcidol per day in the daily group. The therapeutic end point was parathyroid hormone level < 200 pg/ mL. The iPTH levels during the study were monitored, and parameters representative of calcium and phosphate metabolism and side effects were also observed. RESULTS: One hundred and fifty-eight patients were initially enrolled, 91 in the pulse therapy group and 67 in the daily therapy group. There was no significant difference in age, hemodialysis duration, proportion of diabetic nephropathy and systemic diseases, proportion of patients who had received active vitamin D therapy previously, mean initial iPTH level (pulse group 570.47 +/- 295.86 pg/mL; daily group 498.33 +/- 207.84 pg/mL), serum calcium, serum phosphate, alkaline phosphatase (AKP), and albumin between two groups. In the pulse therapy group there were more patients with iPTH levels of 500 to approximately 1,000 pg/mL and > 1,000 pg/mL, so stratified analysis according to iPTH level was used. In therapeutic end point, iPTH levels in both groups were significantly lower compared with those before therapy (pulse group 261.29 +/- 234.97 pg/mL, P < .01; daily group 262.17 +/- 274.82 pg/mL, P < .01). After 4 weeks, the ratio of reaching end point in the pulse group was 35.2%, which was significantly higher than that (19.4%) in the daily group (P < .05). More obvious change was seen in the 200 to approximately 500 pg/mL subgroup by stratified analysis (P < .05), whereas there was no significant difference between the 500 to approximately 1,000 pg/mL and > 1,000 pg/mL subgroup (P > .05). At therapeutic end point, the total ratio of reaching end point did not differ between the two groups, and there were no obvious differences between each subgroup. In the iPTH 200 to approximately 500 pg/mL subgroup, mean iPTH%/week in the pulse group was significantly higher than that in the daily group, and no obvious difference was seen in other subgroups. AKP levels decreased significantly in both groups at therapeutic end point (pulse group 98.42 +/- 54.52 vs. 74.21 +/- 30.68 IU/L, P < .01; daily group 103.3 +/- 68.04 vs. 75.40 +/- 34.12 IU/L, P < .01). On the 4th week, AKP level in pulse group (82.39 +/- 35.23 IU/L) was significantly lower than the initial level (98.42 +/- 54.52 IU/L, P < .05), whereas in the daily group there was no difference between each week. The mean serum calcium, phosphate, and [Ca2+] x [P3+] levels in both groups did not change greatly. Nine patients in the pulse group (9.9%) and 8 patients in the daily group (11.9%) suffered hypercalcemia at least once. Persistent hypercalcemia occurred in 8 patients in the pulse group (8.8%) and 9 patients in the daily group (13.4%), but the difference in proportion did not show statistical significance. The serum phosphate in the daily group was higher after the therapy (1.74 +/- 0.36 vs. 1.89 +/- 0.36 mmol/L, P < .05), whereas that in the pulse group remained unchanged. At therapeutic end point, [Ca2+] x [P3+] level in the daily group was higher than that before the therapy (48.04 +/- 11.71 vs. 55.46 +/- 12.66, P < .05), whereas in the pulse group there was no significant difference. Side effects for both groups were minimal and well tolerated. CONCLUSIONS: Alfacalcidol [1alpha-(OH) D3] has good and safe effects on secondary hyperparathyroidism in maintenance hemodialysis patients. The efficacy and early effects of pulse therapy are superior to those of daily therapy in moderate hyperparathyroidism patients. PMID- 15807188 TI - Use of ascorbate-rich dialysate to attenuate oxidative stress in maintenance hemodialysis patients. AB - BACKGROUND: Oxidative stress exists in uremic milieu, particularly in maintenance hemodialysis (MHD) patients, and accounts for certain long-term complications. Yet little is known about whether supplementation of ascorbic acid (vitamin C, or vitC) via extracorporeal circuit has substantial effects on minifying oxidative impairment. SUBJECTS AND METHODS: The entire experiment consisted of three sections: 1) Practicing ascorbate dialysate among 8 MHD patients in a single dialysis session, compared with a conventional hemodialysis session and another one with intravenous injection of vitC. In each session, oxidative stress markers -namely, plasma total ascorbic acid (TAA), ratio of dehydroascorbic acid (DHAA) to TAA (DHAA/TAA), vitamin E (vitE), and malondialdehyde (MDA)--in both plasma and erythrocytes were measured. 2) A relatively long-term application of ascorbate dialysate in 12 of 23 MHD patients, who were randomly allocated to experimental group (n = 12), and control group (n = 11). Oxidative stress markers and main hematological and biochemical indices were determined at the beginning and end of the period. 3) Application of ascorbate dialysate in 10 MHD patients with intravenous iron treatment, performed in similar procedures as section 1. In addition to determining the aforementioned oxidative stress markers, area under the curve (AUC0-180 min) of ratio of plasma MDA to cholesterol (MDA:Cho) was calculated to evaluate the extent of lipoperoxidation. RESULTS: 1) Plasma TAA gradually decreased during dialysis, whereas a mild increase appeared in MDA. A protruding TAA concentration peak, as well as an extreme DHAA/TAA reduction, followed the injection of vitC, but soon a precipitous fall in DHAA/TAA ensued. Stable plasma TAA and slightly raised vitE were observed when applying ascorbate dialysate. 2) Plasma TAA augmented (27.4 +/- 13.3 vs. 16.8 +/- 9.5 mg/dL, P < .05) and plasma low-density lipoprotein (oxLDL) became two-thirds of baseline data (32.6 +/- 25.2 vs. 83.8 +/- 56.5 micromol/L, P < .05) in the experimental group, whereas oxLDL in the control group reduced quantitatively but not significantly in statistics. (3) As iron sucrose was infused, the decline of TAA and ascending of MDA would be abated not only by intravenous drop of vitC, but also by ascorbate dialysate; however, TAA or MDA curve manifested totally distinguished in the two modalities. AUC0-180 min in ascorbate dialysate group was significantly less than that in control group (400.25 +/- 28.54 vs. 487.25 +/ 109.82). CONCLUSION: Plasma ascorbic acid diminished a great deal during hemodialysis, and at the same time oxidative stress formed and intensified, which will be exacerbated by a remedy of frequent intravenous iron. Ascorbate supplementation, by means of either infusion or extracorporeal circuit, can lessen the loss and therefore attenuate oxidative stress. The latter pattern takes the advantage of retaining the approximate internal balance instead of exquisite change in vivo due to administration of intravenous vitC. PMID- 15807189 TI - Association of cardiac valve calcification and inflammation in patients on hemodialysis. AB - BACKGROUND/AIMS: This study investigates the possible relationship between inflammation and cardiac valve calcification (VC) in patients on hemodialysis (HD), and identifies risk factors for VC in this patient group. METHODS: Seventy nine patients on HD (mean age, 52.2 +/- 13.6 years; mean HD duration, 46.8 +/- 34.3 months) were assessed echocardiographically for the presence of VC. Systolic and diastolic blood pressure (BP) values were determined. The blood parameters studied in each case were hemoglobin, blood urea nitrogen, creatinine, calcium, phosphate, calcium-phosphorous (Ca x P) product, albumin, alkaline phosphatase, intact parathyroid hormone, total cholesterol, high-density lipoprotein, low density lipoprotein, triglyceride, lipoprotein(a), fibrinogen, and C-reactive protein (CRP). The number of patients receiving vitamin D and calcium-containing phosphate binder was determined from records, and presence of diabetes mellitus was noted. RESULTS: Cardiac VC was detected in 36 patients (46%). Five of these patients (6%) had mitral VC, 11 (14%) had aortic VC, and 20 (25%) had calcification of both valves. The patients with VC were significantly older than those without VC (60 +/- 11 vs. 43 +/- 15 years, respectively; P=.001). Compared with the group without VC, the group with calcification had significantly higher systolic (145.1 +/- 14.7 vs. 124.3 +/- 20.7 mmHg, P=.001) and diastolic BP (91.3 +/- 10.3 vs. 75.09 +/- 14.9 mmHg, P=.001); significantly higher phosphate (5.1 +/ 1.4 vs. 4.5 +/- 1.4 mg/dL, P=0.04), CaxP product (48.6 +/- 16.2 vs. 39.8 +/- 11.8, P=.01), lipoprotein(a) [28 (15, 45) vs. 16 (5,42) mg/dL, P=.04], fibrinogen (4.2 +/- 1.2 vs. 3.5-0.9, P=.005), and CRP levels [9 (4, 19) vs. 5 (3, 11) mg/L, P=.05]; and significantly longer HD duration [49 (27, 99) vs. 26 (17, 52) month, P=.01). Apart from age, duration of HD, systolic and diastolic BP, and Ca x P product, VC was associated with CRP (odds ratio, 1.151; P=.007) and fibrinogen (odds ratio, 1.119; P=.005). CONCLUSIONS: The results confirm well-known risk factors for cardiac VC in HD patients, such as older age, longer HD duration, elevated BP, and high Ca x P product. In addition, they suggest that elevated levels of CRP and fibrinogen were associated with VC in the HD population. PMID- 15807190 TI - Chronic renal disease in Kuwaiti nationals: a prospective study during the past 4 years. AB - Our study is a prospective one conducted at Al-Amiri Hospital and including all new cases of chronic renal disease (CRD) seen at the capital area of Kuwait between 1 January 1999 and 30 December 2003. Diagnosis of CRD was based on clinical, laboratory, and radiological features. Kidney biopsies were done when indicated. A total of 271 cases of chronic renal failure (CRF) were diagnosed, of whom 143 were women. The median age was 40 years (range, 5 to 80 years; mean +/- SD: 40 +/- 14). The most common cause of CRF was glomerulonephritis (32%), of which systemic lupus erythematosis and vasculitis constituted 5% and 4%, respectively. Diabetic glomerulosclerosis was the second leading cause of CRD (24%), followed by tubulointerstitial disease (11%) and nephroangiosclerosis (10%). Less frequent causes included renovascular/ischemic disease (6%), obstructive nephropathy (3%), and adult polycystic kidney disease (3%). One hundred and seven patients had 121 incidents of acute deterioration of underlying renal disease. This was mostly due to drugs (22%), infection (21%), and volume depletion (13%). Antiinflammatory drugs were the most common drugs (63%) responsible for the acute decline in renal function. By the end of the study, 18 (7%) patients died, 55 (20%) required maintenance dialysis, and 40 (15%) had received a kidney allograft. Diabetic patients did not differ from nondiabetic with regard mortality, although had more renal replacement therapy (p = .002). Using the Cox regression model, analysis of the relative risk factors likely to contribute to mortality, viz. age, gender, original kidney disease, fitness for transplantation, and mode of presentation, did not show significant factors except for less hazard to death in those diagnosed early with CRD (i.e., on routine testing; relative risk 0.06, p = .01). In conclusion, our study indicates that early diagnosis and management of CRD can improve the patient's quality of life and decrease the cost of frequent hospitalization, morbidity, and even mortality associated with end-stage renal disease. PMID- 15807191 TI - Relation of interdialytic water retention with apparent bicarbonate space, HCO3-, and pH in hemodialyzed uremic patients. AB - This study determines the relationship between interdialytic water retention (IWR) and acid-base homeostasis in uremic patients under regular hemodialysis (HD). To this aim, in 33 regular bicarbonate HD sessions of 11 uremic patients (three HD sessions of 1 week for each patient), blood samples were received from arterial line immediately pre- and post-HD anaerobically in heparinized syringes and the HCO3-, pH, and pco2 were determined. Also in the studied HD sessions, the IWR was estimated and the apparent bicarbonate space percentage (ABS%) pre- and post-HD was calculated by Fernandez et al. (Eq. 1). The mean +/- SD values pre-HD (ABS% = 54.15 +/- 1.49, HCO3- = 18.54 +/- 2.0 mmol/L, pH = 7.32 +/- 0.02, pco2 = 35.44 +/- 3.10 mmHg) and post-HD (ABS% = 49.88 +/- 0.6, HCO3- = 26.33 +/- 1.6 mmol/L, pH = 7.44 +/- 0.02, pco2 = 37.69 +/- 3.00 mmHg) show metabolic acidosis pre-HD and slight alkalosis post-HD. There was a significant positive correlation between IWR and ABS% pre-HD (r = 0.650, p < .0001) and post-HD (r = 0.655, p < .0001), but a significant negative correlation between IWR and HCO3- pre-HD (r = 0.502, p < .003) and post-HD (r = -0.700, p < .001), as well as between IWR and pH pre-HD (r = -0.516, p < .002) and post-HD (r = -0.377, p < .03). In addition, there was a significantly negative correlation between IWR and pco2 post-HD (r = 0.656, p < .001), but not pre-HD (r = 0.0136, PNS). The significantly positive relationship between IWR and ABS% pre- and post-HD, in combination with the significantly negative correlation between HCO3- and pH pre- and post-HD, indicates that the IWR negatively influences the acid-base homeostasis in hemodialysis patients without residual renal function, and may worsen the cardiovascular physiology and tissue oxygenation of these patients. PMID- 15807192 TI - Hypochlorous acid and 3,4-dihydroxyphenylalanine increase the formation of serum protein lipofuscin-like fluorophores in vitro. AB - INTRODUCTION: Serum protein lipofuscin-like fluorophores (LLFs) that include fluorescent advanced glycation end products (AGEs), are an index of protein modification and levels are abnormally high in hemodialysis patients. To investigate the possibility that hypochlorous acid (HOCl) and 3,4 dihydroxyphenylalanine (DOPA) may contribute to high LLFs concentrations, we have examined the effect of these factors on serum protein LLF formation in vitro. METHODS: Protein LLF concentration was measured at excitation 350 nm and emission 460 nm and was expressed in arbitrary units relative to quinine sulphate fluorescence. Oxidation of serum or other solutions with HOCl was carried out at room temperature for 30 minutes and serum was delipidated before measurement of protein LLFs. RESULTS: Serum protein LLF concentration increased non-linearly by a maximum 247% with increasing HOCl concentration in the range 6.5-32.9 mmol/L and this was mirrored by a decrease in protein tryptophan fluorescence. HOCl (32.9 mmol/L) increased LLFs in human gamma-globulin solutions (15-fold in 12 mg/mL and 5-fold in 60 mg/mL solutions) and did not alter LLFs appreciably in human serum albumin solution (60 mg/mL). Addition of DOPA (265 micromol/L) significantly (P<0.001) increased LLF formation in serum by nearly 2-fold during 3 days incubation under air. CONCLUSIONS: These data suggest that HOCl and DOPA are capable of generating serum protein LLFs and that gamma-globulins appear to be an important substrate for protein LLF formation in human serum. These findings may be relevant to the abnormally high concentrations of serum protein LLFs and impaired immune response in hemodialysis patients. PMID- 15807193 TI - Theory, targets and therapy in systemic lupus erythematosus. AB - The treatment of systemic lupus erythematosus (SLE) has been refined over the years, with the recognition that a fine balance lies between aggressive and prompt therapy and attendant complications brought upon by immunosuppressive therapy itself. However, there has been limited change to the repertoire of drugs available to treat this challenging disease. The current standard therapy for severe manifestations of SLE includes the use of high-dose corticosteroids and cytotoxic agents such as cyclophosphamide (CYC), which have been associated with an increased risk of serious and opportunistic infections. The need for safer, more targeted therapies has been recognized and now, with the exponential increase in the understanding of immunopathogenic mechanisms in SLE, the way has been paved for the development of biologic or targeted therapies in SLE. Although the potential immunosuppression, long-term safety issues and cost-effectiveness remain unclear. These targeted therapies may range from small molecules that specifically inhibit inflammatory processes at an intracellular, cell-cell or cell-matrix level to monoclonal antibodies, soluble receptors or natural antagonists that interfere with cytokine function, cellular activation and inflammatory gene transcription. PMID- 15807194 TI - Anti-cytokine therapy in systemic lupus erythematosus. AB - In the course of the disease, a wide variety of cytokines is dysregulated, many of which likely influence systemic lupus erythematosus (SLE) autoimmunity and/or lupus tissue inflammation. Proinflammatory cytokines in particular, such as TNF, IL-6, IL-18 or IFN-gamma, may play a major role in propagating the inflammatory processes responsible for tissue damage. These cytokines are overexpressed both systemically and locally, and preliminary results from open-label trials and/or animal studies suggest potential benefits of blocking either of these inflammatory mediators. Since new therapeutic agents may soon offer many ways to influence the process, controlled clinical trials following open-label safety studies are of central importance to arrive at optimized therapies for SLE patients. PMID- 15807195 TI - Statins: immunomodulators for autoimmune rheumatic disease? AB - Inhibitors of 3-hydroxy-3-methylgluttaryl coenzyme A (HMG-CoA) reductase, or statins, are used extensively to reduced elevated lipid levels and reduce cardiovascular risk. However, accumulated evidence suggests that stains not only act by lowering cholesterol levels, but also exert pleiotropic effects on many essential cellular functions including cell proliferation, differentiation, and survival and participate in the regulation of cell shape and motility. Thus cardiovascular benefit is provided by lowering raised cholesterol levels and by modulation of the inflammatory component of this disease. Such an anti inflammatory effect may also benefit patients with autoimmune rheumatic disease. This overview assesses the evidence for using statins in patients with rheumatoid arthritis and systemic lupus erythematosus (SLE). PMID- 15807196 TI - Block and tackle: CTLA4Ig takes on lupus. AB - Blockade of antigen nonspecific costimulatory signals is a promising approach for the treatment of autoimmune diseases including systemic lupus erythematosus (SLE). CTLA4Ig, an antagonist of the CD28/B7 costimulatory interaction, effectively prevents SLE onset in several murine models and, when used in combination with cyclophosphamide, can induce remission of active SLE nephritis. In this review we describe the known mechanisms of action of CTLA4Ig both in normal immunity and in autoimmune disease models and address issues about its activity that still need to be resolved. We discuss the preclinical use of CTLA4Ig in murine SLE models and the rationale for a clinical trial in SLE patients. PMID- 15807197 TI - BLyS antagonists and peptide tolerance induction. AB - The major impediment to drug development for systemic lupus erythematosus (SLE) is its heterogeneity. The unpredictable manner in which lupus targets different organs at varying intensity makes the study of new drugs and the optimization of their administration extremely difficult. With the advent of novel, targeted biologic agents for SLE, it can be hoped that more strategic, lupus-relevant immune modulation will lead to safer and more effective treatments. Two alternative new approaches to lupus treatment are reviewed. The first involves selective inhibition of a single protein (BLyS), which may play a central role in host defense and in the pathogenesis of SLE. Although this approach is finely targeted to the inhibition of a single protein which is known to be upregulated in SLE patients, the positioning of BLyS at a critical hub in the immune response suggests that more global adverse repercussions on immunity might still occur. The second strategy is the use of peptides designed to specifically induce tolerance in limited autoreactive immune responses. Immune repercussions might, at least in theory, be almost nonexistent with this kind of approach. Concerns that limited induction of tolerance might have equally limited impact on the complex immune disorder of SLE are not borne out in preliminary murine data. Specific development programs are ongoing using both of these strategies and have recently entered human trials. PMID- 15807198 TI - Anti-B cell therapy (rituximab) in the treatment of autoimmune diseases. AB - The use of B cell depletion as a mode of treatment for non-Hodgkin's lymphoma was first utilized in 1997 when Rituximab, a chimeric human-mouse monoclonal antibody which has a high affinity to the CD20 antigen expressed on B cells, became available. Over 500000 lymphoma patients have been treated worldwide with this drug and it has a good safety record. The notion that B cells might be critical to the development of rheumatoid arthritis led to the extension of the use of B cell depletion to this condition and a recent double blind controlled trial has shown very encouraging results. In addition, B cell depletion either using Rituximab alone, or in combination with cyclophosphamide and corticosteroids has also been reported to have been of great benefit in some patients with severe systemic lupus erythematosus albeit in open label studies. This review considers the mechanism of action of the drug, the clinical trials that have been reported, and tries to place its current use in patients with autoimmune rheumatic disease in context. PMID- 15807199 TI - [Therapeutic effects of phytosterols and phytostanols in cholesterolemia]. AB - Plant sterol and stanol esters are called "functional" compounds due to their hypocholesterolemic properties. The objective of this review is to update recent findings concerning the effect of phytosterols in the blood cholesterol, emphasizing the results from experimental and human studies. The hypocholesterolemic effect is observed with the intake of 2.5g/day of phytosterols or phytostanols. Daily intake, usually of stanols, for 4 weeks has shown to to be effective in lowering blood total- as well as LDL-cholesterol by about 10%. The mechanism of action in lowering blood cholesterol comes from their structural similarity to cholesterol, hence they act by competing with cholesterol at the luminal absorption site. The adverse effects of a high intake of phytosterols and phytostanols are the lower absorption of some liposoluble vitamins and antioxidants. PMID- 15807200 TI - [Postprandial lipemia in Colombian young adults from different ethnic groups]. AB - Postprandial lipemia has been associated with atherogenesis and other non infectious chronic diseases. A descriptive, non aleatory study of 51 healthy young adults (23.8 +/- 4 years) of different ethnic background was carried out to identify possible personal or life style factors associated with the response of plasma lipids after a mixed carbohydrate and lipid load. Personal, family, life styles including use of drugs and activity and ethnic background were recorded. Anthropometrical measurements, a short insulin and postprandial lipemia tests were done. From these observations body mass index (BMI) and area under the curve (AUC) for lipids and glucose were calculated. High mean values were observed in AUC for glucose, triacylglycerol and nonesterified fatty acids (NEFA). Postprandial glucose with differences by age (p=0.05 Bonferroni) but no ethnic, gender, BMI or insulin sensitivity related significant differences were found. Mestizos (40%) showed two, and Negroes three (27%) postprandial triglyceridemia peaks. Noteworthy, subjects presenting four peaks had elevated BMI. Higher triglyceridemia values were found in white and mestizos (p<0.05), in subjects older than 29 and in males (p<0.05). Statistical correlation (r2=0.70) between BMI and triglyceridemia and change in postprandial trigliceridemia was found in white and black subjects. NEFA declined up to the second hour, but at 6-7 hours they reached levels higher than base values. Higher than reported values for UCA and BMI and an abnormal response to fat intake with an early release of NEFA was found. The high correlation between BMI and postprandial trigliceridemia suggests that body mass index could be a good predictor of postprandial trigliceridemia. PMID- 15807201 TI - [Effect of a high fat or high carbohydrate breakfast on postprandial lipid profile in healthy subjects with or without family history of type 2 diabetes mellitus]. AB - The objective of this study was to evaluate the effect of a high fat or high carbohydrate breakfast on postprandial lipid profile in healthy subjects with or without family history of type 2 diabetes mellitus. A single blind, controlled clinical trial with parallel groups was performed in 20 healthy subjects; 10 subjects with family history of type 2 diabetes mellitus and 10 individuals without that background. Each group was randomized to receive a high fat or high carbohidrate breakfast. A metabolic profile that included fasting and postprandial lipids, as well as, the assessment of insulin sensitivity were performed. Lower high-lipoprotein cholesterol (p < 0.02) and apolipoprotein A1 (p < 0.03) concentrations were found in subjects with family history of type 2 diabetes mellitus than those without that background. In this same above mentioned group with the high carbohydrate breakfast, there were significant increments in apoliprotein B at minute 300 (p < 0.03) and in triglycerides at minute 360 (p < 0.03). In the group without family history of diabetes that received the high fat breakfast, there were increments in triglycerides (p < 0.03) and very-low density lipoprotein concentrations at minute 180 (p < 0.03). In conclusion, healthy subjects with family history of type 2 diabetes showed some atherogenic characteristics in their metabolic profile, and the high carbohydrate breakfast produced in them increments in apolipoprotein B and in triglycerides, meanwhile that, in those subjects without such background the high fast breakfast produced unfavorable effects on their lipid concentrations. PMID- 15807202 TI - [Predictive accuracy of body mass index in estimating body fatness measured by bioelectrical impedance]. AB - The aim of this study was to determinate the performance of body mass index (BMI) for the diagnosis of obesity and as a predictor of body fatness in adult Chilean subjects. We conducted a study in 433 women (18-73 years old; BMI: 19.7 to 69.7 kg/m2) and 264 men (18-83 y.; BMI: 19.1 to 54.8 kg/m2). Bioelectrical resistance was measured by impedance method and fat mass percent (FM%) was calculated by fatness-specific equations developed by Segal et al. Obesity was defined as a BMI of at least 30 kg/m2. Increased fatness was defined by the FM% cut-off points of at least 25% for men and at least 30% for women. Sixty-four percent of women and 23.6% of men with BMI below 30 kg/m2 had FM% higher than 30% and 25%, respectively. A value of BMI of 26 kg/m2 in women and 30 kg/m2 in men had the best agreement to the cut-off points of fatness according to sensitivity vs. specificity analysis The following equations were developed to predict individual fatness: women FM% = 0.96 x BMI + 0.154 x age + 1.44 (r2 = 0.75; standard error 3.8%); men FM% = 0.99 x BMI + 0.141 x age - 9.914 (r2 = 0.66; standard error 4.4%). Differences between measured and predicted FM% presented a wide variation, with a range of +/- 2 sd of 7.5% in women and 8.8% in men. The commonly used value of BMI 330 kg/m2 as a cut-off point for obesity does not have adequate sensitivity and specificity for the screening of increased fatness subjects, specially in women. In this study BMI shows a low reliability as a predictor of individual body fatness, particularly in men and in subjects with a BMI below 30 kg/m2. PMID- 15807203 TI - [Influence of nutritional status on the effectiveness of a dietary supplement of live lactobacillus to prevent and cure diarrhoea in children]. AB - The influence of undernutrition on the effectiveness of a dietary supplement of live lactobacillus regarding prevention and treatment of infantile diarrhoea was studied on 200 children 6 to 24m old. Children, undernourished (D) or controls (C), received for 90 days, in a blind experiment, a fermented milk providing L. Acidofilus y L. Casei (10(7)-10(8)/ml) (LB) or an equivalent amount of fluid milk (L). diarrhoea episodes were recorded and classified according duration: 1-4, 5 14 days, and protracted diarrhea (DP) those lasting beyond 14 days. Study requirements were fulfilled by 119 children: D-L: n=25; D-LB: n=32; C-L: n=27 y C LB: n=35. Preventive aspects were evaluated through number of episodes and through their lasting the therapeutic ones. Episodes recorded were 134: 29 in D L; 48 in D-LB; 37 in C-L and 20 in C-LB. In the D-LB group over 90% episodes were very short (1-4 days), percentage far higher to that observed in D-L and similar to C-LB. There were 12 episodes of DP, 9 D and 3 in C, all in the L groups. In conclusion, although the fermented milk prevent half episode in the controls but not in the undernourished, it was able to shorter episodes duration and prevent protracted diarrhoea irrespectively of nutritional status. Therefore, undernutrition impaired the ability of the lactobacillus supplement to prevent children diarrhoea, which implicates immune system, but not curative effects which are the result of local actions. PMID- 15807204 TI - [Evaluation of the effect of probiotic cultures added to commercial yogurt over a known population of Listeria monocytogenes and Escherichia coli O157:H7]. AB - The effect of probiotic cultures over Listeria monocytogenes and Escherichia coli O157:H7 during yogurt storage was evaluated. Two different yogurt brands, one with additional probiotic cultures (Lactobacillus casei and L. acidophilus) were inoculated with known populations (106 UFC/g) of either L. monocytogenes or E. coli O157:H7 in three different times and stored for 32 days at 5 degrees C. Every four days the count of lactic bacteria, the added pathogens and pH was evaluated, according to the methodology described in the Bacteriological Analytical Manual. The pH and lactic bacteria population were constant during the testing period. Yogurt with additional probiotic cultures reduced the population of L. monocytogenes in 8 days, the population of E. coli O157:H7 in 16; yogurt with no additional probiotics took 20 days to reduce L. monocytogenes to non detectable levels and even after 28 days of storage, E. coli O157:H7 was cultured. In this work, the beneficial effects of additional probiotic cultures in yogurt is confirmed again. PMID- 15807205 TI - [Evaluation of the effect of probiotic cultures on two different yogurt brands over a known population of Staphylococcus aureus and the production of thermonuclease]. AB - The effect of probiotic cultures over known populations of Staphylococcus aureus inoculated in yogurt was studied; also the production and stability of its thermonuclease during yogurt storage was evaluated. In three different occasions, two different yogurt brands, one with additional probiotic cultures (Lactobacillus casei and L. acidophilus), were inoculated with known populations of S. aureus in high and low concentration (10(9) CFU/g and 10(7) CFU/g), respectively. These samples were stored for 28 days at 5 degrees C. Every four days the count of lactic bacteria, S. aureus and pH were evaluated, according to the methodology described in the Compendium of Methods for the Microbiological Examination of Foods, Vanderzant & Splittstoesser. The presence of thermonuclease was determined using petrifilm for S. aureus from 3M company. The pH and lactic bacteria population were constant during the testing period. Yogurt with additional probiotic cultures (high and low concentration) lowered the population of S. aureus to non detectable levels in 8 days; but, S. aureus could be cultured from yogurt without probiotics even after 24 days of incubation. Same time, the presence of thermonuclease was positive in all tests; it was not affected by probiotics. The presence of thermonuclease is related to the production of S. aureus enterotoxin. This work emphasizes again the beneficial effects of probiotic cultures in yogurt over bacteria and the importance of keeping hygienic practices in order to avoid the contamination of food with S. aureus and the eventual production of its enterotoxin, since it is not affected by probiotics. PMID- 15807206 TI - [Evaluation of the presence of bacteria in food and environment of an Oncological Service of a National Hospital, San Jose, Costa Rica]. AB - The microbiological quality of 92 samples from the Oncological Service of a National Hospital, San Jose, Costa Rica was evaluated during the months of February to July, 2002. Of these samples, 48 were from salads, refreshments, and fruits and 44 from surfaces, air and hands of the medical personnel. The presence of total and fecal coliforms was determined by the total plate count technique, using Violet Red Bile Agar for the isolation of total and fecal coliforms, Baird Parker Agar for Staphylococcus aureus, Cetrimide Agar for Pseudomonas and Listeria was enriched in University of Vermont Broth, Fraizer Broth and isolated in Oxford Agar. From the food samples analyzed, 77% were positive for at least one of the parameters studied; the fruits presented the greater contamination (94%) and the cooked salads the minor, even though Listeria was isolated in 13% of them. All tested salads (100%) were positive for fecal coliforms. Pseudomonas and S. aureus were isolated from fruits, raw salads, and refreshments. All the analyzed surfaces, with an exception, were negative for the parameters evaluated. 75% of the hands of the medical personnel were positive for at least one of the parameters evaluated; 25% had total coliforms (2.8x10(3) UFC/hand), 13% fecal coliforms (2.4x10(3) UFC/ hand), 69% S. aureus (1.3x10(3) UFC/hand) and 19% Pseudomonas sp. 115 isolates were randomly selected, and identified using miniaturized tests (API). When comparing these identifications with the 166 isolations done at the Clinical Laboratory of the Hospital from the infected patients of oncology, by means of variance analysis, it was determined that both bacterial populations were statistically similar. PMID- 15807207 TI - [Microbiological and physicochemical evaluation of pasteurized beverages fortified with orange deodorized residues extracts]. AB - Microbiological and physicochemical parameters of pasteurized beverages conditioned with aqueous extracts from orange deodorized residues were evaluated. The fruits used were selected according to following criterion: homogenous maturity, without physical damage and absence of apparent chlorophyll. Orange peels were dried and transformed to flour. The juice was evaluated by means of these parameters: pH 3.90, degrees Brix 10, titrable acidity 0.33 g of citric acid/100 mL and total carotenoids 0.0078 mg/mL. Volatil compounds of the flour that may to cause bitterness were separated by means of two methods of deodorization: distillation in current of vapor and in autoclave to 121 degrees C; then, the flour was extracted with water (relation 1:50 p/v). Pasteurized citric beverages (orange juice) were elaborated adding the deodorized extracts. pH, degrees Brix, titrable acidity and total carotenoids showed no significant differences (P>0.05). Microbiological results were according to pasteurized products. Sensorial analysis by untrained panel showed no signiificant differences. In conclusion, the deodorant processes were effectives and permitted the inclusion of aquasoluble compounds as flavonoids with antioxidant activity. PMID- 15807208 TI - [Development of bakery products for greater adult consumption based on wheat and rice flour]. AB - The present investigation was developed as a contribution to Guatemalan's elderly food and nutrition. Its main objective was to evaluate the chemical, nutritional and sensory quality of bread prepared from the partial substitution of wheat flour with rice flour. Wheat flour substitutions with rice flour in the order of 15, 20, 30, 40, 50 and 60% were evaluated. Differences with the control (100% wheat bread) were found during the process of preparation, as well as texture, volume, height, weight and specific volume. Important effects in dough handling were noted specifically in the 40, 50 and 60% rice bread. Thus, a sandy texture was found in breads of higher rice levels. The bread protein quality increased with the level of substitution; however the protein quality difference between the wheat bread and the bread with 60% rice flour did not achieve statistical significance. Based on a statistical analysis of the physical properties the bread with 30 and 40% rice flour was selected, and through a preference test between these last two, the 30% rice flour bread was selected as the sample best suited to the present study's purposes. This bread was not different to wheat bread in many nutritional parameters, although in others it showed to be superior. Each serving size of bread has a weight of 80 grams (2 slices) that contributes adequate quantity of calories, protein and sodium, although a little less dietary fiber than 100% wheat bread. PMID- 15807209 TI - [Nutritional characterization of carbohydrates and proximal composition of cooked tropical roots and tubers produced in Costa Rica]. AB - Tropical roots and tubers (TRT) are important staple foods in the tropics. TRT are produced by small farmers and have received only limited attention. The objective of this paper was to evaluate nutritionally important carbohydrate fractions and proximal composition of cooked cassava (Manihot esculenta), cocoyam or tannia- (Xantosoma sp) and yam (Dioscorea alata) grown in Costa Rica. Twenty boiled samples of each TRT were analyzed for proximal composition, following AOAC protocols. Carbohydrate fractions (dietary fiber, available and resistant starch) and alpha-amylolysis rate were assessed by enzymatic methods. All TRT analyzed consisted mainly of water and carbohydrates, representing moderate dietary fiber and energy sources. They show a low protein level and contain no fat. Moisture, protein, ashes and dietary fiber contents vary significantly (p<0.01) among the three species. In terms of their proximal composition, these locally grown TRT differ from TRTs cultivated in the South Pacific area, but appear similar to other Latin American varieties. In all samples most of the dietary fiber (84-88%) is insoluble; available starch ranges between 74 and 84% of the dry matter, whereas resistant starch content varied between 0.7 and 1.7%. Amylolysis rate was similar for cassava and cocoyam, whereas a significantly slower digestion (p<0.01) was recorded for yam. Compared to values reported in the literature for other starchy foods and to gelatinized potato starch, used as reference sample, the studied TRT showed intermediate amylolysis rates. It is concluded that consumption of these TRT may be promoted in Costa Rica and other countries with a similar nutritional situation. PMID- 15807210 TI - [Lipidic pattern of 25 Mexican marine fishes with special emphasis in their n-3 fatty acids as nutraceuticals components]. AB - The aim of this study was to characterize and to evaluate the lipidic composition of mexican marine fishes with special emphasis in n-3 fatty acids as nutraceuticals. The edible portion of 25 species: humidity (H), crude protein (CP), total lipids (TL) and fatty acids (FA). The average content (g/100g edible portion) of H was 75.20, PC was 18.40, TL was 3.60. Four n-3 FA were identified in all the samples and they were found in the next abundance order (mg/100g edible portion): C22:6n-3 (DHA)(229.60), C20:5 n-3 (EPA)(52.10), C18:3 n-3 (ALA)(11.80) and C20:3 n-3 (2.25). By their origin and climate there were no difference. By their biologycal classification, n-3 FA content was higher in bony fishes than cartilaginous fishes. It was detected a proportional relation with the n-3 FA concentration and total lipid content. According to their ecotic distribution there were numerical differences in DHA content (mg/100g edible portion) between pelagics (420.70), benthopelagics (125.30) and demersals fishes (225.40). Fatty fishes had higher content of EPA and DHA (mg/100g edible portion) (109.27 and 552.72) than semifatty fishes (56.12 and 226.29) and leanness (15.95 and 96.52), respectively. Bony, fatty and pelagic fishes had a higher content of EPA+DHA. According with the international recommendation values (200 to 600 mg EPA+DHA/day) the 44% of the analyzed species could be considered as functional foods due to their high content of EPA + DHA in a range of 220 to 1300 mg/100g. PMID- 15807211 TI - [Characterization of microbial population present in the edible seaweed, Monostroma undulatum, Wittrock]. AB - The microbiological quality of Monostroma undulatum, Wittrock from the Southern Argentinean coast, was studied for its application for human food. Also the diversity and function of the native bacterial population to this green seaweed was analyzed. Samples were collected in Puerto Deseado, province of Santa Cruz, Southern Argentina (47 degrees 45'L.S., 65 degrees 55'L.W). The samples were analyzed for the presence of psycotrophic heterotrophic bacteria, marine heterotrophic bacteria, low nutritional request bacteria (LNRB), marine low nutritional request bacteria (LNRB marine), Vibrio spp, total and thermotolerant colifom bacteria, anaerobic sulfite reducing bacteria, yeasts and moulds. The isolates were identified using standard techniques based on morphologic, physiologic and metabolic characteristics. Among the gram-negative bacteria isolated, the predominant genera belonged to Vibrio (20%), E. coli inactiva (18%), Flavobacterium (11%), Flexibacter (9%), Moraxella (9%), Alcaligenes/Pseudomonas group (9%), Aeromonas (2%), Acinetobacter (2%). Cotophaga (2%), Photobacterium (2%), Ps/Caulobacter/Alteromonas/Spirillum group (2), The main genus of gram-positive bacteria was Staphylococcus. Human pathogenic bacteria were not detected. Fecal contamination indicator bacteria were not isolated from fresh seaweed and seawater. These results showed an adequate microbiological quality of seaweed acceptable for human food. The bacterial population associated to Monostroma undulatum, consisted of gram-negative, marine and psycotrophic microorganisms, including vibrios and enterobacteria as their main components. Also the identified bacteria showed a great capacity to hydrolyze different substrates and so they might contribute to the balance of this marine ecosystem. PMID- 15807212 TI - Mineral content of the honey produced in Zulia state, Venezuela. AB - The mineral content of the honey produced in five zones of the Zulia state, Venezuela, during dry and rainy seasons was determined. The analyzed elements were: sodium, potassium (by emission spectroscopy), calcium, magnesium, copper, iron, manganese (by atomic absorption spectroscopy), phosphorus (phosphate ions, by colorimetric method), and ash content of raw honey samples directly collected from different beekeepers. The mean values for Na, K, Ca, Mg, Cu, Fe, Mn, and P were 353+84; 1774+138; 237+66; 52+24; 0.76+0.43; 13.5+10.23; 0.92+0.42 and 1642+323 mg/kg respectively. The mean ash content was 0.431+0.15%. Potassium was the most abundant of the elements determined. This results confirm that Zulian honey can be considered a good source of minerals. PMID- 15807213 TI - [Biochemical and chemical partial characterization of Bauhinia forficata Link seeds]. AB - Seeds of Bauhinia forficata species were submited to biochemical characterization concerning fatty acids analysis, protein fractionization, and hemaglutinanting activity. The seed elementary analysis showed a high protein and lipids contents with 21.24% and 19.45% respectively. The more abundant fatty acid was linoleic acid with 46.47% of the lipidic fraction. With the exception of prolamins, the different proteic fractions (albumin, globulins, acid and basic glutelins) showed hemaglutinanting activity against rabbit red cells no treated and treated with proteolitic enzymes. The fraction acid glutelin showed the higher specific hemaglutinanting activity (1072.25 UH/mg P) against rabbit blood pre-treated with trypsin. Glutamin (16.20%) and Valin (11.07%) were the more abundant amino acids in the seeds. Therefore, B. forficata represent a possible optional source of food because exhibit a high energetic values. PMID- 15807214 TI - To publish or not to publish--that is the question. PMID- 15807216 TI - A reduction in intraovarian arterial blood flow resistance after ovulation is necessary to achieve pregnancy in natural cycle. AB - PURPOSE: Color Doppler imaging permits the accurate localization of vessels and high-frequency pulsed Doppler ultrasonography has improved the resolution of flow velocity waveforms. In this study, intraovarian arterial blood flow before and after follicle rupture in the natural cycle was examined using transvaginal color flow Doppler imaging and changes in intraovarian arterial resistance in relation to the outcome of infertility treatment was analyzed. METHODS: In a prospective study, 227 spontaneous cycles in 118 infertile patients who were undergoing infertility treatment at the division of Reproductive Medicine in our center were recruited in this study. The impedance to flow in intraovarian vessels was measured by means of transvaginal color flow Doppler imaging during the periovulatory period in the natural cycle of all patients. The pulsatility index (PI) of intraovarian arterial blood flow and pregnancy rate was evaluated. RESULTS: On the basis of PI values before and after follicular rupture, 227 cycles were classified into severely decreased (113 cycles) and not-severely decreased groups (114 cycles). The pregnancy rate per cycle in the severely decreased group was 18.6% (21/113), significantly higher than that in the not severely decreased group (7/114; 6.1%, p = 0.004). The miscarriage rate was similar in the two groups. CONCLUSIONS: A reduction in intraovarian blood vessel resistance is necessary to achieve pregnancy in a natural cycle. PMID- 15807217 TI - Do lunar phases affect conception rates in assisted reproduction? AB - OBJECTIVES: To assess the effect of the phases of the moon on pregnancy rates in humans following in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment. DESIGN: Retrospective, observational study. SETTING: Reproductive Medicine Unit, Liverpool Women's Hospital. PATIENT: Complete data for all women undergoing assisted conception procedures over a period of 13 years (1995-2002). INTERVENTION: Assisted conception procedures--IVF and ICSI. MAIN OUTCOME MEASURES: Biochemical pregnancy that is positive pregnancy test result following embryo transfer. RESULTS: There was no significant effect of any lunar phase on the incidence of biochemical pregnancy (p-value 0.71). Age of the woman significantly affects the chances of pregnancy, (OR 0.95, 95% CI 0.91, 0.998, and p-value 0.04). The chances of pregnancy rises significantly with increase in the number of embryos replaced from 1 to 2 (OR 2.97, CI 1.36, 6.48, and p-value 0.01). CONCLUSION: Pregnancy rates in humans, following assisted conception, appears to be independent of the effect of the lunar phase during which embryo transfer is carried out. PMID- 15807215 TI - Abdominal ultrasound-guided embryo transfer improves clinical pregnancy rates after in vitro fertilization: experiences from 330 clinical investigations. AB - PURPOSE: To investigate the effect of ultrasound-guided embryo transfer on the rate of implantation and clinical pregnancy. METHODS: A prospective randomized trial was performed to compare ultrasound-guided embryo transfer with the traditional method. A total of 330 patients were randomly divided into two groups on the day of embryo transfer. For the cases (n = 178), ultrasound-guided was used; controls (n = 152) was performed using routine methods. RESULTS: The rate of implantation and clinical pregnancy for the cases (19.6 and 37.1%, respectively) was significantly higher than for the controls (12.6 and 25%, respectively; p < 0.05). CONCLUSION: Ultrasound-guided embryo transfer can significantly increase the rate of implantation and clinical pregnancy, and should be recommended as a routine procedure in the process of in vitro fertilization and embryo transfer (IVF-ET). PMID- 15807218 TI - In vivo assessment of the regulation of transforming growth factor alpha, epidermal growth factor (EGF), and EGF receptor in the human endometrium by medroxyprogesterone acetate. AB - PURPOSE: The present study evaluated the in vivo effect of medroxyprogesterone acetate (MPA) on the localization of immunoreactive transforming growth factor alpha (TGFalpha), epidermal growth factor (EGF), and their common receptor (EGF R) in the human endometrium. METHODS: The study design was a randomized clinical trial enrolling 36 healthy women with regular menstrual cycles. The participants were randomly assigned into three groups: groups 1 (n = 11) and 2 (n = 17) received placebo and were submitted to endometrial biopsy during the proliferative and secretory phases of menstrual cycle, respectively; group 3 (n = 8) received MPA (10 mg/day) for 10 days followed by endometrial biopsy, which was performed during the secretory phase. Immunohistochemistry was used to localize TGFalpha, EGF, and EGF-R in the endometrial tissue. RESULTS: TGFalpha was present markedly in the luminal and glandular epithelia but also in the periglandular stroma, with a distribution pattern similar in the three experimental groups. EGF immunostaing was equally distributed in epithelial and stromal layers of the endometrium and remained unchanged in endometrial samples from women treated with MPA compared to placebo. EGF-R was expressed only in the epithelium. The intensity of EGF-R immunostaining was higher in secretory than in proliferative endometrium and was further increased by administration of MPA (p < 0.05, chi square test). CONCLUSION: The present results suggest that the progestogen induced in vivo differentiation of secretory endometrium does not require dramatic changes in the expression of EGF or TGFalpha, whereas EGF-R may be up regulated. PMID- 15807219 TI - Preliminary experience with low concentration of granulocyte-macrophage colony stimulating factor: a potential regulator in preimplantation mouse embryo development and apoptosis. AB - PURPOSE: To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the development of preimplantation mouse embryos. METHODS: Mouse 2-cell embryos were collected and cultured in P-1 medium supplemented with GM-CSF at different concentrations. Using reverse transcription-polymerase chain reaction, expression Bcl-2 and Bax mRNA in blastocyst were evaluated in the GM CSF group and control group. Apoptosis detection was performed using the in situ apoptosis detection kit in mouse blastocysts. The statistical significance of the data was analyzed using t-test and chi-square test. RESULTS: The development of blastocyst increased to 89% in the addition of GM-CSF (0.125 ng/mL), compared to controlled group (80%). The number of cells staining for apoptosis was lower in GM-CSF group than that in the control group. Bcl-2 expression was found to be upregulated in blastocysts in the GM-CSF supplemented group compared to the control group. CONCLUSION: These results suggest that GM-CSF might be an important regulator in embryo development. PMID- 15807220 TI - Successful pregnancy after the vitrification of zygotes using commercial vitrification solutions and conventional straws to protect against infections in liquid nitrogen. AB - PURPOSE: To report on successful birth after the transfer of postthawed human zygotes that were vitrified using a conventional straw for the purpose of protecting them from infections and a low-toxicity cryoprotectant that is commercially sold. METHODS: A primary infertile couple presented at our IVF program. After being checked for fertilization, the embryos were not transferred to the uterus at that cycle. Instead, all of them were cryopreserved at the 2 pronuclei stage using our original vitrification method. After the vitrification and warming of four zygotes, two embryos were transferred into the uterus. RESULTS: Twenty-one 2-pronuclei embryos were vitrified in liquid nitrogen. After 2 embryos were thawed and transferred, successful pregnancy was the outcome, and a healthy boy was born at term. CONCLUSIONS: Vitrification is a simple procedure and requires less time than slow freezing. Vitrification of zygotes in a conventional straw seems to be sufficient for viability and works to store the zygotes safely. PMID- 15807221 TI - Successful pregnancies in patients with estrogenic anovulation after low-dose human chorionic gonadotropin therapy alone following hMG for controlled ovarian hyperstimulation. AB - OBJECTIVE: To demonstrate that folliculogenesis can be sustained with 200 IU human chorionic gonadotropins (hCG) after FSH-priming and result in pregnancy in women with estrogenic ovulatory dysfunction and risk factors for severe ovarian hyperstimulation syndrome (OHSS). DESIGN: CASE REPORT: Three women with infertility associated with estrogenic ovulatory dysfunction and hyperinsulinemia who appeared to be at high risk for severe OHSS during gonadotropin therapy. INTERVENTIONS: After 10 days of receiving either 150 IU hMG or recombinant FSH, patients were switched to 200 IU hCG/day alone for 2-3 days. 5,000 IU of hCG was then administered followed by either home intercourse, intrauterine insemination or transvaginal oocyte retrieval-embryo transfer. MAIN OUTCOME MEASURES: Endovaginal ultrasound measurement of follicle number and size, serum estradiol levels, symptoms of ovarian hyperstimulation, pregnancy test, and evaluation of pregnancy by transvaginal ultrasound. RESULTS: After discontinuation of hMG or recombinant FSH, serum estradiol concentrations continued to rise, and follicles >14 mm continued to grow during low-dose hCG administration. All women conceived without developing symptoms of OHSS. Pregnancy outcomes achieved include a term singleton delivery, a term twin delivery, and triplets delivered at 31 weeks gestation. CONCLUSION: The use of low-dose hCG alone is sufficient for supporting the late stages of folliculogenesis in women with estrogenic ovulatory dysfunction. This ovulation induction regimen appears to support the follicular growth of larger follicles while decreasing the number of smaller preovulatory follicles, thereby reducing a known risk factor for OHSS. We report on the positive pregnancy outcomes in 3 women with estrogenic ovulatory dysfunction and clinically appeared to be at high risk for developing severe OHSS who safely underwent this protocol. PMID- 15807222 TI - Reflections and comments on regulations of biotechnologies that touch the beginning of human life. PMID- 15807223 TI - The gambling follow-up scale: development and reliability testing of a scale for pathological gamblers under treatment. AB - OBJECTIVE: To provide preliminary data on the Gambling Follow-Up Scale (GFS), a new scale assessing recovering gamblers. Secondary goals included assessing the impact of "work status," "family relationship," "leisure," and "enrolment in Gamblers Anonymous (GA)" on gambling (all items from the scale), together with the impact of treatment. METHOD: Using the GFS, 3 independent raters interviewed gamblers under treatment. The sample was collected in 2 university centres in the city of Sao Paulo, Brazil. Patients attended psychotherapy coupled with psychiatric follow-up, participation in GA, or both. RESULTS: We interviewed 47 pathological gamblers; 13 were interviewed twice, with a minimum interval of 6 months, for a total of 60 GFS interviews. Interviews took on average 6.0 minutes, SD 2.7. Interrater concordance ranged from 82% to 95% (intraclass correlation coefficient range 0.85 to 0.99, P < 0.001). A factorial analysis showed a 1 factor solution (Eigenvalue = 2.4, 47.6% of total variance accounted). "Leisure," "frequency and time gambling," and "family relationship" showed considerable loadings (0.84; 0.71; 0.71),whereas "enrolment in GA" and "work status" showed moderate loadings (0.59; 0.56). A linear regression model significantly correlated gambling (R2 = 0.356; P < 0.001) with "leisure" and length of treatment. Treatment modalities affected "leisure" (F2,43 = 5.00, P = 0.011), with GA attendees reporting more regular and gratifying activities. CONCLUSIONS: The GFS showed interrater reliability and construct validity. More leisure and lengthier treatment were significantly relAted to less gambling. GA enrolment seemed to particularly benefit the quality of leisure. Future studies could profit from the quickness and simple structure of the GFS in providing shareable outcome measures. PMID- 15807224 TI - The relation between perceived need for mental health treatment, DSM diagnosis, and quality of life: a Canadian population-based survey. AB - OBJECTIVES: Prevalence estimates of mental disorders were designed to provide an indirect estimate of the need for mental health services in the community. However, recent studies have demonstrated that meeting criteria for a DSM-based disorder does not necessarily equate with need for treatment. The current investigation examined the relation between self-perceived need for mental health treatment and DSM diagnosis, with respect to quality of life (QoL) and suicidal ideation. METHODS: Data came from an Ontario population-based sample of 8116 residents (aged 15 to 64 years). The University of Michigan Composite International Diagnostic Interview was used to diagnose mood, anxiety, substance use, and bulimia disorder according to DSM-III-R criteria. We categorized past year help seeking for emotional symptoms and (or) perceiving a need for treatment without seeking care as self-perceived need for treatment. We used a range of variables to measure QoL: self-perception of mental health status, a validated instrument that measured well-being, and restriction of activities (current, past 30 days, and long-term). RESULTS: Independent of subjects' meeting criteria for a DSM-III-R diagnosis, self-perceived need for treatment was significantly associated with poor QoL (on all measures) and past-year suicidal ideation. CONCLUSIONS: Self-perceived need for mental health treatment, in addition to DSM diagnosis, may provide valuable information for estimating the number of people in the population who need mental health services. The relation between self perceived need for treatment and objective measures of treatment need requires future study. PMID- 15807225 TI - Perceived quality of life in patients with bipolar disorder. Does group psychoeducation have an impact? AB - OBJECTIVE: A large body of research has now accumulated concerning quality of life (QoL) for patients with major depressive disorder, both in terms of describing levels of well-being and in terms of assessing the impact of treatment interventions. However, there is little information concerning QoL for patients with bipolar disorder (BD), and there is relatively little published evidence concerning the effectiveness of psychological interventions for BD. We aimed to assess the impact of a time-limited psychoeducation (PE) group therapy upon perceived QoL among patients with BD. METHOD: Participants were patients (n = 57) with BD type I or II who were clinically described as euthymic or mildly symptomatic. Treatment intervention was a standardized, 8-week group PE course delivered in a mood disorders program in British Columbia, Canada. Using retrospective chart review and the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q), we assessed QoL at baseline and at 8 weeks. RESULTS: Mean baseline Q-LES-Q scores were 56%, representing moderate impairment in QoL. Group PE was associated with a 5-point increase in Q-LES-Q scores (where higher scores indicate better QoL). Examination of the questionnaire's subscales revealed that 2 domains (that is, physical functioning and general satisfaction) increased significantly following PE, with the remaining domains showing nonsignificant trends toward improved functioning. Multivariate analysis indicated that only one factor (having had a recent episode of depression) significantly predicted pre- and posttreatment Q-LES-Q scores. CONCLUSION: Patients with BD continue to show impaired QoL even when clinically euthymic. Although preliminary, our results show that group PE is associated with improved QoL in this population, both in terms of general satisfaction and in relation to levels of physical functioning. The use of PE as an adjunct to pharmacotherapy in BD should be further studied with particular emphasis on characterizing the effects of treatment intervention on perceived QoL. PMID- 15807226 TI - [Depersonalization--current data]. AB - OBJECTIVE: Depersonalization is a fascinating clinical phenomenon referring to a self-consciousness disorder, characterized by emotional detachment from one's own feelings, thoughts, or actions. This article intends to summarize the current literature in this area. METHOD: Using the Medline data base, we reviewed literature addressing the clinical, etiology, nosology, physiopathology, and treatment of depersonalization. CONCLUSIONS: Derealization means that perception of the world and of external reality are altered. These 2 phenomena are often associated. They are not specific to any psychiatric entity and are reported in many different psychiatric syndromes. Many factors, including use of different substances, are involved in their onset. The physiopathology is still little known. However, some conceptual models suggest partial amygdala inhibition combined with activation of other amygdaloid structures. A serotoninergic functioning impairment is indicated in different pharmacologic studies. Different psychotropic drugs, especially serotoninergic antidepressants, have been proposed for pharmacotherapy; however, there are no conclusive randomized studies, and the contribution of psychotherapy in treating these patients is still questioned. PMID- 15807227 TI - Qualitative research in psychiatry. AB - This paper is an overview of qualitative research and its application to psychiatry. It is introductory and attempts to describe both the aims of qualitative research and its underlying philosophical basis. We describe the practice and process of qualitative research and follow this with an overview of the 3 main methods of inquiry: interviews, focus groups, and participant observation. Throughout the paper, we offer examples of cases where qualitative research has illuminated, or has the potential to illuminate, important questions in psychiatric research. We describe methods of sampling and follow with an overview of qualitative analysis, appropriate checks on rigour, and the presentation of qualitative results. The paper concludes by arguing that qualitative methods may be an increasingly appropriate methodology to answer some of the demanding research questions being posed in 21st century psychiatry. PMID- 15807228 TI - Finding our way: an introduction to path analysis. AB - Path analysis is an extension of multiple regression. It goes beyond regression in that it allows for the analysis of more complicated models. In particular, it can examine situations in which there are several final dependent variables and those in which there are "chains" of influence, in that variable A influences variable B, which in turn affects variable C. Despite its previous name of "causal modelling," path analysis cannot be used to establish causality or even to determine whether a specific model is correct; it can only determine whether the data are consistent with the model. However, it is extremely powerful for examining complex models and for comparing different models to determine which one best fits the data. As with many techniques, path analysis has its own unique nomenclature, assumptions, and conventions, which are discussed in this paper. PMID- 15807229 TI - Suicidality in adolescents and adults with fetal alcohol spectrum disorders. PMID- 15807230 TI - Absent dose-response in the posttraumatic stress disorder symptoms of 350 Holocaust survivors. PMID- 15807231 TI - Sex of the offender, sex of the victim, and motivation in filicidal situations in Quebec. PMID- 15807232 TI - Seizures, coma, and coagulopathy following olanzapine overdose. PMID- 15807233 TI - Posttraumatic mood disorder: a new concept. PMID- 15807234 TI - Proinflammatory cytokines: a common denominator in depression and somatic symptoms? PMID- 15807235 TI - [Effect of N, P, K on rhizome yield and diosgenin content of Dioscorea zingiberensis]. AB - OBJECTIVE: A field experiment was conducted to investigate the different effects of various applying ratios of nitrogen (N), potassium (K) and phosphate (P) on diosgenin content and Rhizoma yield of Dioscorea zingiberensis. METHOD: Experiment was conducted with both one factor and orthogonal design, and data was analyzed statistically on SPSS 11. 5. RESULTS: Compared with the control groups,the yield of rhizome was significantly increased by N (P < 0.01), K (P < 0.01) , P (P <0.05). The content of diosgenin has no obvious increase (P > 0.05) by individual application of N, P and K, while changed significantly (P < 0.05) by the combination of proper levels of N, P, K. CONCLUSION: The degree of effect on diosgenin content was ranged with N > K > P, while on output of dioscorea rhizome was K > N > P, and the combination of N, K, P2 was superior to others. PMID- 15807236 TI - [Study on tissue culture technique of Liriope spicata]. AB - OBJECTIVE: To study the tissue culture technique of Liriope spicata. METHODS: The tender stem segments with roots as explants were cultured on various media with different kinds of hormones, a lot of bud clusters were induced and grew into seedlings. RESULTS: A great quantity of whole aseptic seedlings can be formed quickly by the tender stem segments on the media. PMID- 15807237 TI - [LC-MS analysis of three kinds of Mutong medicinal materials]. AB - Three kinds of Mutong medicinal materials (Lardizabalaceae, Ranunculaceae, Aristolochiaceae) were analyzed qualitatively by LC-MS. The results show Mutong of Lardizabalaceae and Mutong of Ranunculaceae have more the same chemical constituents, and Mutong of Aristolochiaceae has less the same constituents with them. These results are consistent with their medicinal properties and explain the similarity and difference in medicinal properties from chemical basis. PMID- 15807238 TI - [Morphological and microscopical identification of Murraya kwangsiensis]. AB - OBJECTIVE: To identify medical plant Murraya kwangsiensis by its tissue characters. METHOD: Morphological characters and microstructure of this plant were described and illustrated. RESULT: Oil cell, fiber cell and calcium oxalate prism were found in its tissue. CONCLUSION: Tissue characters can be taken into account of developing basis for quality standard of this medical plant. PMID- 15807239 TI - [Re-study on characteristic fingerprint of volatile oil from Herba Pogostemonis by GC]. AB - OBJECTIVE: To establish the characteristic fingerprint and parameter of volatile oil from Herba Pogostemonis. METHODS: The dffferent collecting time and county samples of Herba Pogostemonis were determined by GC. RESULTS: Accuracy, stability and repeatability of the method were good. The same region samples had good stability and main constituents were same in different collection time, but content were different. The samples collected from different region were more different in constituents and content. CONCLUSION: The 11 components in common buildup the characterisitic fingerprint of volatile oil from Herba Pogostemonis. The ratiio of pogostone to patchoulic alcohol were used index parameter for quality evaluation and characteristics of producing region. PMID- 15807240 TI - [Study on chemical constituents from Swertia kauitchensis Franch]. AB - OBJECTIVE: To study the major chemical constituents from the whole herb of Swertia kauitchensis Franch. METHOD: Compounds were separated and purified with silica gel column and microreticular resin, and their structures were elucidated on the basis of spectroscopic evidence (UV, IR, MS, NMR). RESULT: Ten compounds were identified as oleanolic acid (1), beta-sitosterol (2), daucosterol (3), gentiacaulein (4), bellidifolin (5), methylswertianin (6), swertianin (7), 1,7 dihydroxy-3,4,8-trimethoxyxanthone (8), swertianolin (9),1,3,5,8 tetrahydroxyxanthone (10). CONCLUSION: All compounds were isolated from Swertia kauitchensis Franch for the first time. PMID- 15807241 TI - [Extraction of ferulic acid from enzymatic hydrolysate by ion-exchange adsorption]. AB - Anion-exchange resin, D201 was used to prepare ferulic acid from the enzymatic hydrolysate of wheat-bran and the influence of adsorption and desorption operation was researched. The optimal technology is as follows: under the room temperature, the concentration of ferulic acid in the extraction solution is 2000 mg/L to approximately 3000 mg/L, pH and flow velocity were kept around 9.0 and 1 ml/min respectively; elution solution is composed of ethanol: water: HCl in ratio 60:36:4 (v/v), and the velocity of elution is 1 ml/min. Under optimal conditions,the recovery percent of ferulic acid from enzymatic hydrolysate exceed 97% and the purification production is in evidence. PMID- 15807242 TI - [Purification and characteristics of polypeptide with inhibition of platelet aggregation from Agkistrodon blomhoffii Bioe]. AB - A polypeptide, SV-PP-2, was purified from Agkistrodon blomhoffii Bioe by HPLC C18 column. The SV-PP-2 has molecular weight of 23339.00 Da, and can inhibit the ADP induced platelet aggregation in dose-dependent manner. There have been no such polypeptides reported yet which has inhibition of platelet aggregation and similar molecular weight to SV-PP-2. PMID- 15807243 TI - [Isolation and identification of alkaloids from Armeniaca mume Sieb]. AB - OBJECTIVE: To study the alkaloids from Armeniaca mume Sieb. METHODS: The alkaloids were extracted with chloroform from 80% alcoholic extract and concentrated. Then they were transferred into aqueous solution and absorbed by macroporous cation exchange resin. The concentrated total eluent was extracted with chloroform. The alkaloids were purified by recrystallization with petroleum ether and sublimation. Then they were determined by Fourier Transform Infrared spectroscopy, Mass spectrometry and Nuclear magnetic resonance etc methods. RESULT: Two alkaloids were separated and their structures were elucidated as 2,2,6,6-tetramethyl-4-piperidone and tert-butylurea on the basis of spectral evidence. CONCLUSION: They are new alkaloids from A. mume. PMID- 15807244 TI - [Research of anti-proliferation of curcumin on A549 human lung cancer cells and its mechanism]. AB - OBJECTIVE: To investigate the mechanism of anti-tumor effects of curcumin on human lung cancer cell (A549). METHOD: MTT colorimetry method, fluoroscope, FCM combine PI and Annexin V-FITC double pigmentation method and Western blot method were used. RESULT: Under the effect of the curcumin, cell grew against the wall and suspended in the culture liquid, the A549 cell nucleolus were found fragmentated into different size of apoptosis body under fluoroscopy. The cell proliferation were obvious suppressed after treated with different concentration curcumin for 72 hours. The IC50 were 18 micromol/L by using linear regression. The apoptosis induced by curcumin of A549 cell is concentration dependent. With curcumin increased from 5 micromol/L to 30 micromol/L, Annexin-FITC single positive cell (early apoptosis cell) increased from 3.4% to 59.1%. When curcumin concentration reached 40 micromol/L, PI and Annexin V-FITC double positive cell (secondary apoptosis necrosis cell) became major part of cells, and the cell showed G2 phase block. Observed with western blot method, with the increase of curcumin concentration to 10 micromol/L, the expression of PARP increased simultaneously. CONCLUSION: Curcumin can interfere cell growth cycle of A549 cell and suppress cell growth. The suppression effect is concentration dependent. The effect depends not only from the nonspecific cytotoxic but also from induced cell apoptosis. PMID- 15807245 TI - [Effects of Peritrophe roxburghiana on blood pressure in renal hypertensive and hyperlipidemic rats]. AB - OBJECTIVE: To observe the effects of Peristrophe roxburghiana (HSX) on the blood pressure in renal hypertensive and hyperlipidemic rats (RHHR), and investigate the possible mechanisms of its pharmacological effects. METHODS: The two-kidney, two-clip method was used to produce the renovascular hypertensive rats, then high lipid emulsion was administered to produce the model of hyperlipidemic rats. Drugs were administered for 5 weeks. Blood pressure was measured weekly before and after administration. After rats were killed at the end of the 5th week,blood was sampled to measure plasma angiotensin IT and serum NO. Angiotensin II in the thoratic aorta was measured too. RESULTS: The BP decreased significantly to the end of 5th week (P < 0.05) after treatment of 1-2 weeks with HSX. HSX (H or L dosage) increased the level of serum NO evidently at the end of the experiment. No significant statistical difference was found in the level of plasma angiotensin, but HSX(H) and captopril decreased the value of angiotensin II in blood vessel (P < 0.01). CONCLUSION: HSX decreased the blood pressure significantly for RHHR and the mechanism of its antihypertensive effect was possibly related to the increasing of serum NO and the decreasing of angiotensin II1 in the thoratic aorta. PMID- 15807246 TI - [Effects of exopolysaccharide from Tolypocladium sinense on murine immunocytes in vitro]. AB - OBJECTIVE: To elucidate the effects of exopolysaccharide from Tolypocladium sinense on murine immunocytes in vitro. METHODS: Extracted exopolysccharide (EPS) from cultured Tolypocladium sinense. The effects of EPS on mouse immune parameters were detected in vitro using cell culture techniques. RESULTS: The results indicated that EPS might significantly not only promote mouse macrophage neutral red uptake ability and cytotoxicity (P < 0.05), but also stimulate their thymus lymphocytes proliferation (P < 0.05). CONCLUSION: EPS may elevate murine cellular immunity to a certain. PMID- 15807247 TI - [The effect of "Jian Pi Dao Zhi Fang" on human gastric carcinoma cells]. AB - OBJECTIVE: To study the effect of Traditional Chinese medicine "Jian Pi Dao Zhi Fang" (JPDZF) with invigorating the spleen and purgative function on the growth of gastric carcinoma cells and the expression of VEGF and P53 in gastric carcinoma cells. METHODS: Gastric carcinoma cells were cultured in the RPMI-1640 media with 10% serum containing JPDZF for 48 hours. The cell cycle were tested with flow cytometer. Expression of P53, VEGF and its receptor Fit were evaluated by immunohistochemistry. RESULT: The cell cycle of gastric carcinoma cells cultured in the media with JPDZF were different from the control. The expression of P53, VEGF, Flt decreased with the dosage increasing of JPDZF. CONCLUSION: Traaditional Chinese medicine JPDZF can inhibit the growth of gastric carcinoma cells and the expression of P53, VEGF, Flt. PMID- 15807248 TI - [The effect of borneol on level of HA and 5-HT in rat's hypothalamus]. AB - OBJECTIVE: To observe the effect of borneol on level of HA and 5-HT in rat's hypothalamus. METHODS: The levels of HA and 5-HT in rat's hypothalamus were determined before and after p.o. with different amount of borneol by high performance liquid chromatography with electrochemical detection (HPLC-ECD), and the borneol's effect on the levels of HA and 5-HT was also studied. RESULTS: The level of HA in rat's hypothalamus after different dose of borneol were higher than before administration. The level of HA in 20 min group after middle dose increased significantly comparing with before administration (P < 0.01), the others of group after middle dose, the 45 min group after high dose, the 20 and 45 min after low dose were also increased significantly than before administration (P < 0.05). After different dose of borneol, the level of 5-HT in rat's hypothalamus changed as follows: the level of 5-HT after high dose were higher than before administraton (P <0.05 or P < 0.01 ); the level of 5-HT after 5, 20 and 45 min of middle and low dose incrased significantly (P <0.05 or P < 0.01). CONCLUSION: Borneol could increase the levels of HA and 5-HT in rat's hypothalamus. PMID- 15807249 TI - [Study on technique of extraction and purifying chlorogenic acid in Eucommia ulmoides Oliver leaves]. AB - Ethanol was used as solvent to extract chlorogenic acid from leaves of Eucommia ulmoides Oliver leaves. The optimum conditions are obtained by response surface analytical method. Optimized conditions for the extraction of chlorogenic acid were obtained: ethanol 51.7%, extraction temperature 55.8 degrees C, ratio of material to liquor 1:12.8, extraction time 2 h. The static adsorption tests are conducted among six kinds of macroporous resins to select NKA-II resin as a optimum resin to adsorb the chlorogenic acid in Eucommia ulmoides Oliver leaves. The optimum adsortion-elution conditions are as follow: adsorption flow rate is 2 ml/min, crude liquid flew twice through the adsorption column,the optimum adsorbent is 40% ethanol. PMID- 15807250 TI - [Study on preparation of the inclusion compound of curcumin with beta cyclodextrin]. AB - OBJECTIVE: The inclusion compound of curcumin with, beta-CD was preparated to increase the solubility and bioavaliability of curcumin. METHODS: Prepare inclusion compound was studied with orthogonal design. The inclusion compound was verified with fluorometry, TLC, and phase solubility. RESULT: The best way to prepare inclusion compound are as follows: the molar ratio between curcumin and, beta-CD is 1:1, temperture is 40 degrees C, reaction time is 2 h. CONCLUSION: Solubility of inclusion compound is 10 times than curcumin. PMID- 15807251 TI - [Study on the extraction process of polysaccharide from Lycium barbarum]. AB - OBJECTIVE: To choose the optimum extration process of polysaccharide from Lycium barbarum L. METHODS: The optimal water extration process is selected with the orthogonal design. The yield and the content of polysaccharide were used to evaluate the factor levels. RESULTS: The ideal extraction process is: the polysaccharide is extracted with water for 3 times,with solvent volume 8 times amount each time (10 times amount for the first time), extraction time 1 h each time and stired 10 min each hour. CONCLUSION: The experimental results provide the basis for the water extration process of polysaccharide from Lycium barbarum L. PMID- 15807252 TI - [Determination of sennoside A in rhubarb extracts by HPLC]. AB - OBJECTIVE: To determine sennoside A in rhubarb extracts by solvent extraction and SFE-CO2. METHODS: Sennoside A was extracted by ultrasonic vibration. The analytical column was Allitima RP C18 (250 x 4.6 mm, 5 microm). The mobile phase consisted of 0.1% TFA-HCN (44:56). The flow rate was 1.0 ml/min. The UV detector wavelength was 280 nm and the reference wavelength was 600 nm. The column temperature was 25 degrees C. RESULT: Considering the extracting rate of sennoside A, solvent extraction was better than SFE-CO2. PMID- 15807253 TI - [Expression of xenogeneic homologous epidermal growth factor receptor ectodomain in Pichia pastoris]. AB - OBJECTIVE: To develop a secreted expression system of xenogeneic homologous epidermal growth factor receptor (EGFR) ectodomain using Pichia pastoris (P. pastoris), thus providing a basis for subsequent antitumor immunological researches on EGFR subunit protein vaccine. METHODS: In order to construct the secreted expression vector pPICZaA-mer and pPICZaA-her, we recombined the cDNA of xenogeneic (human) EGFR ectodomain (her) and the corresponding control mouse EGFR ectodomain (mer) with the empty plasmid pPICZaA, respectively. Then the recombinant plasmids were linearized by Sac I and transformed into P. pastoris strain GS115 by electroporation. The positive transformants were selected on YPDS plate including Zeocin and were induced to express by methanol. Consequently, the proteins in the culture supernatant were assayed with SDS-PAGE and Western-blot. So we could choose the high-level secreted expression recombinant strain to produce protein vaccine vastly. RESULTS: We got the secreted expression of mouse and human EGFR ectodomain in P. pastoris. The relative molecular mass of the two aimed proteins is about 95 x 10(3). The Western-blot analysis demonstrated that the expression proteins have much good antigenicity and specificity. CONCLUSION: The recombinant plasmids pPICZaA-mer and pPICZaA-her could be induced to express the EGFR aimed proteins by methanol in methanol-trophic yeast expression system. PMID- 15807254 TI - [Effect of hypoxia/reoxygenation (H/R) on expression of MICA and MICB in human hepatocytes]. AB - OBJECTIVE: It has been found that the human major histocompatibility complex class I chain-related antigen A,B (MICA,B), the ligand for NKG2D (an activating receptor of NK cell, CD8+ cell and gammadelta T cell), could be induced by cellular stress. The purpose of this study is to investigate whether Hypoxia/Reoxygenation could upregulate expression of MICA,B gene in human hepatocytes (HL-7702). METHODS: The model of cell hypoxia/reoxygenation was used to mimic ischemia/reperfusion injury (IRI) of graft organ. The human hepatocytes were exposed to hypoxic condition for 15 h under 1% O2, 94% N2 and 5% CO2, then reoxygenated for 2 h, 4 h, 6 h, 12 h and 24 h separately under normal condition. The levels of MICA and MICB mRNA were measured by semi-quantitative RT-PCR. The surface expression of MIC were detected by immunofluorescence with a rabbit polyclonal antibody which is recommended for the detection of MICA,B of human origin simultaneously. RESULTS: Hpoxia/Reoxygenation increased the levels of MICA,B in hypoxic hepatocytes. CONCLUSION: In this article is firstly reported that H/R could increased the levels of MICA,B in human hepatocytes, yet the meachinsm is unknown. The upregulated expression of NKG2D ligand possibly activates NK cells and is hence associated with transplantation. PMID- 15807255 TI - [Construction of co-expression plasmid containing tPA and VEGF165 genes and its expression in VSMCs]. AB - OBJECTIVE: To construct the eukaryotic co-expression plasmid of tissue plasminogen activator (tPA) genes and vascular endothelia growth factor165 (VEGF165) and observe its expression in vascular smooth muscle cells (VSMCs). METHODS: The tPA and VEGF165 genes were cloned from hominal heart tissue with RT PCR, and then the tPA and VEGF165 genes were cloned into eukaryotic expression plasmid pBudCE4.1 to construct the eukaryotic co-expression plasmid pBudCE4.1/tPA VEGF165. The pBudCE4.1/tPA-VEGF165 was transfected into VSMCs via lipofection mediation. The expression levels of tPA and VEGF165 mRNAs in the transformed VSMCs were detected by RT-PCR and the expression levels of tPA and VEGF165 proteins were detected by ELISA. RESULTS: The size of RT-PCR product of tPA and VEGF165 genes was 1.9 kb and 576 bp respectively. Restrictive enzyme digestion analysis showed that recombinant co-expression plasmid pBudCE4.1/tPA-VEGF165 had been constructed successfully. The expression of tPA and VEGF165 at mRNA and protein levels in the transformed VSMCs and cells culture supernatant were detected respectively by RT-PCR and ELISA. CONCLUSION: The recombinant eukaryotic co-expression plasmid pBudCE4.1/tPA-VEGF165 has been successfully constructed. The tPA and VEGF165 are expressed in transformed VSMCs. PMID- 15807256 TI - [Antitumor efficacy of CD/5-FC suicide gene therapy system mediated by Bifidobacterium infantis against melanoma]. AB - OBJECTIVE: To observe the antitumor efficacy of cytosine deaminase/5 fluorocytosine (CD/5-FC) suicide gene therapy system mediated by Bifidobacterium infantis against melanoma in vitro and in vivo. METHODS: Bifidobacterium infantis transfected by CD gene was incubated with 5-FC in anaerobic condition. The supernatant of bacteria was collected and added to the melanoma B16-F10 cells for assessment of the killing effect of this suicide gene system. Mice were incubated with melanoma B16-F10 cells to establish animal models. Then the mice bearing melanoma were treated by a combination of Bifidobacterium infantis transfected by CD gene and 5-FC. Bifidobacterium infantis transfected by CD gene was injected into the tail vein of mice and 5-FC was given introperitoneally. RESULTS: (1) In vitro, the tumor cells in test groups appeared damaged with remarkable changes in morphology, and the cell growth was significantly inhibited as compared against those in the control group. (2) In animal experiment, after the mice melanoma model being treated with the recombined bacteria and 5-FC for 21 days, the tumor growth span was remarkably prolonged and the tumor volume was significantly inhibited when compared with control. The differences became more obvious with the passage of observation time. CONCLUSION: CD/5-FC suicide gene therapy system mediated by Bifidobacterium infantis was noted to have good antitumor effect on melanoma. PMID- 15807257 TI - [The construction of recombinant plasmid with prostate-specific membrane antigen promoter controlling reporter gene expression]. AB - OBJECTIVE: To study the specificity of prostate-specific membrane antigen (PSMA) promoter in controlling gene expression. METHODS: PSMA promoter gene was amplified with PCR, and then the promoter gene was cloned into the vector pEGFP-1 to construct a recombinant plasmid, which was transfected into different cell lines such as LNcap, PC-3, MCF-7, A549. Green fluorescent protein (GFP) expression was observed. RESULTS: The recombinant plasmid constructed, and PSMA promoter uniquely showed modulating activity in PSMA positive cell line. CONCLUSION: PSMA promoter possesses PSMA positive cell specificity, as well as prostatic tissue specificity. PSMA promoter may have the potential for use in targeted gene therapy of prostate adenocarcinoma. PMID- 15807258 TI - [Construction and expression of recombinant plasmid with prostate-specific membrane antigen promoter and enhancer regulating uracil phosphoribosyltransferase gene]. AB - OBJECTIVE: To construct the recombinant expression plasmid pPSMA(EP)-UPRT including UPRT gene that is regulated by PSMA(enhancer/promoter). METHODS: By use of PCR, UPRT gene was amplified from E. coli JM109 genome. Then UPRT gene was cloned into the recombinant expression plasmid pPSMA(enhancer/promoter)-EGFP that is driven by prostate-specific membrane antigen promoter and enhancer. RESULTS: It was found that EGFP was digested by two restriction enzymes from the recombinant expression plasmid pPSMA(enhancer/promoter)-EGFP, and UPRT gene was linked with the recombinant expression plasmid by T4 DNA ligase. We succeeded in constructing the recombinant expression plasmid pPSMA(enhancer/promoter)-UPRT. The recombinant plasmid sequences were verified, and the expression in vitro was measured by MTT. CONCLUSION: The recombinant expression plasmid pPSMA(enhancer/promoter)-UPRT is regulated targetly by prostate-specific membrane antigen promoter and enhancer. It is, of importance to us in studying the UPRT/5 FU for gene therapy of prostate cancer, especially suicide gene therapy (CD/5-FC system). PMID- 15807259 TI - [Expression of CNTF and PDGF in spared dorsal root ganglion after partial dorsal root rhizotomy]. AB - OBJECTIVE: To investigate the expression of ciliary neurotrophic factor (CNTF) and platelet derived growth factor (PDGF) in spared dorsal root ganglion (DRG) after partial dorsal root rhizotomy. METHODS: Twenty adult cats were divided into four groups. Five cats were kept intact in the control group. Fifteen cats were subjected to bilateral root rhizotomy, and on the 3rd day, 7th day and 14th day after operation, they were sacrificed as subjects in the three experiment groups respectively (n=5 per group) and their DRGs (L6) were taken. Immunochemical ABC method was used to detect the distribution of CNTF and PDGF-immunoreactivity neurons in those DRGs. The quantitative analysis was conducted to get the numbers of CNTF, PDGF-positive total, large, and medium-small sized neurons in each group. RESULTS: CNTF, PDGF-immunoreactants were distributed in large and medium small sized neurons. The numbers of total and medium-small sized CNTF positive neurons were noted to be decreased on the 3rd day after operation (P<0.05), and no difference 7th and 14th days was seen when compared with control (P>0.05), but the large positive neurons showed no difference. The total and medium-small sized PDGF positive neurons were found decreased apparently on the 3rd day and 7th day, but there was no difference on the 14th day as compared with normal level. Large positive neurons displayed no change at every time. CONCLUSION: Partial dorsal root rhizotomy exerts different influence on the expression of CNTF and PDGF for different neurons in spared DRG. PMID- 15807260 TI - [Expression of Bax, Bcl-2 and caspase-3 in spared dorsal root ganglion after partial dorsal root rhizotomy]. AB - OBJECTIVE: Investigating the expression of Bax, Bcl-2 and Caspase-3 in dorsal root ganglion after partial dorsal root rhizotomy in spared root. METHODS: Adult cats were used. The study comprised a normal control group (n=5) and three expriment groups (n=5 per group). The unilateral root rhizotomy was performed on the 15 cats of the 3 experiment groups. On the 3rd, 7th and 14th days after partial dorsal rhizotomy, the DRG (L6) of 5 cats per batch were taken and made into frozen sections and were stained under the same condition using specific Bax, Bcl-2 and Caspase-3 antibody by ABC method. The Bax, Bcl-2 and Caspase-3 positive large-, medium- and small-sized neurons in DRG were counted in the 3 experiment groups and in the normal control group. RESULTS: In the intact animals of the normal control group, the Bax, Bcl-2 and Caspase-3 immunoreactants were mainly distributed in medium, small neurons and a few large neurons. The number of Bax positive neurons in spared DRG of the 3rd day group increased apparently as compared with that of the normal control group but decreased apparently in the 14th day group and showed no significant difference when compared versus control (P>0.05). There was no significant difference between the 14th day group and normal control group. Bcl-2 expression in spared DRG displayed no difference among the four groups. The number of Caspase-3 positive neurons in spared DRG of the 3rd day group increased apparently when compared against control (P<0.05), but it was not significantly different from that of the 7th day and 14th day groups (P>0.05). CONCLUSION: Partial dorsal root rhizotomy mainly has an effect on the expressions of Bax, Bcl-2 and Caspase-3 in medium- and small-sized neurons of spared DRG, the peaks of the expressions may appear 3-7 d after the operation. PMID- 15807261 TI - [Changes in expression of SP in spinal lamina II following hemisected spinal cord injury in rats]. AB - OBJECTIVE: To explore the changes in expression of Substance P (SP) in spinal lamina II at different time following hemisected spinal cord injury (hSCI). METHODS: Twenty Sprague-Dawley rats were randomly divided into intact group and hSCI group at days 3, 7 and 21. The spinal cords were hemisected between T13 and L1. The animals were sacrificed 3, 7, 21 days after operation. The L3 segments were taken out and sectioned continuously into sections (20 microm). The expression of SP was measured by immunohistochemical ABC method, and the number of SP positive varicosities in lamina II was counted. RESULTS: SP positive varicosities were observed in spinal lamina II. Compared with control group, the number of SP positive varicosities on the injured side apparently increased 3 days after hSCI, but decreased 7 days after operation, then recovered to normal by 21 days after operation. Noteworthily, the number of SP positive varicosities on the intact side was higher than that on the injured side. CONCLUSION: SP, as a neurotransmitter of nociceptive information, may be related to the process of hSCI. PMID- 15807262 TI - [IGFBP-2 mRNA expression in gastric cancer tissues and its roles in proliferation, differentiation and peritoneal metastasis of gastric cancer]. AB - OBJECTIVE: To detect the expression of IGFBP-2 mRNA in gastric cancer tissues and its roles in neoplasia and peritoneal metastasis of gastric cancer. METHODS: The IGFBP-2 mRNA expression levels in the fresh tissues from normal mucosa, primary cancer and peritoneal metastasis of 35 patients with gastric cancer were determined by the quantitative real time RT-PCR method, and the samples were sliced up and stained with hematoxylin-eosin for pathologyical examination. RESULTS: The levels of IGFBP-2 mRNA expression in gastric primary cancer tissues were significantly higher than those in the tissues from normal mucosa and peritoneal metastasis (P<0.05). Yet, IGFBP-2 mRNA expression levels showed no difference between the tissues of normal mucosa and peritoneal metastasis (P>0.05). CONCLUSION: The expression of IGFBP-2 mRNA possibly plays an important role in proliferation, differentiation and peritoneal metastasis of gastric cancer, and it might be useful in a new method for clinical prognostication and treatment of gastric cancer peritoneal metastasis. PMID- 15807263 TI - [Changes in the expression of vital cytokines for pancreatic development in early embryo]. AB - OBJECTIVE: To clarify the changes in the expression of vital cytokines for pancreatic development in early embryo and hence provide preclinical data regarding embryonic pancreas transplanted for treatment, of diabetes mellitus. METHODS: Sample collection was conducted in accordance to the principle of informed consent. Histochemical S-ABC method was adopted in studying pancreas at 7-12 weeks of gestation, and analysis was made on vital cytokines expression as well as the differentiation and forming of pancreatic islets. RESULTS: It was found that the expression of Insulin, Glucagon, Somatostatin and Cytokeratin begins at 7 weeks, and the change is consistent with the differentiation and formation of pancreatic islets. IGF-I and F-VIII appear at 12 weeks. CONCLUSION: Insulin, Glucagon, Somatostatin, Cytokeratin, IGF-I and F-VIII play a regulatory role in the development of embryonic pancreas and are related to the differentiation and formation of endocrine and exocrine glands of pancreas. The pancreas after 12 weeks of gestation can be used as a donated graft for pancreas transplantation. PMID- 15807264 TI - [The genotypic profiles of Candida albicans isolates from patients with oral lichen planus]. AB - OBJECTIVE: The aim of this study was to investigate the genotypic profiles of Candida albicans isolates from erosive oral lichen planus (OLP) and non-erosive OLP. METHODS: A total of 112 isolates obtained from healthy control (26), erosive OLP (62) and non-erosive OLP (24) were screened for genotypic profiles by using the randomly amplified polymorphic DNA (RAPD) assay. RESULTS: RAPD analyses with some random primer reavealed 4 different genotypes among all isolates, and there was significant difference in genotypic constitution between every two groups. Statistically, B and D were the major types in healthy group; A and C were the major types in erosive OLP; A and D were the major types in non-erosive OLP. CONCLUSION: Some Candida albicans isolates with special genotypic profiles may contribute to the development and progression of OLP. PMID- 15807265 TI - [Construction and expression of the major outer membrane protein PI of Neisseria gonorrhoeae in Escherichia coli]. AB - OBJECTIVE: To construct Neisseria gonorrhoeae major outer membrane protein PI gene recombinants for expression of the target protein in E. coli. METHODS: Four clinic isolates of Neisseria gonorrhoeae were collected, and then the genome DNA of these strains was extracted. The gene encoding for PI of Neisseria gonorrhoeae was amplified by PCR, inserted into the cloning vector pBS-T; the recombinant plasmids pBS-T-PI-NG were constructed and sequence analysis was performed. Then PI gene fragments were inserted into expression vector pET30b to form pET30b-PI NG recombinants. The PI protein expression was induced by adding IPTG in the inocula. The expressed proteins were analyzed by SDS-PAGE. RESULTS: The pET30b-PI NG expression recombinants for four clinic isolates of Neisseria gonorrhoeae were constructed successfully; three of the four expression recombinants were expressed successfully. CONCLUSION: The expressed PI protein will be applied in the further research for PI antigenicity and immunological activity. This will be very helpful for the further construction of preventive vaccines directed against Neisseria gonorrhoeae infection. PMID- 15807266 TI - [Cloning and expression of superoxide dismutase gene from Deinococcus radiodurans in E. coli]. AB - OBJECTIVE: To construct expressing recombinant of Mn-SOD of Deinococcus radiodurans and express the target protein in E. coli BL21(DE3). METHODS: SOD gene was amplified by PCR from genomic DNA of Deinococcus radiodurans and inserted into expression plasmid pET-30a(+) to create the recombinant pET-SOD. After being analyzed by the restriction endonuclease, the plasmid was transformed into E. coli BL21(DE3), and the recombinant protein was expressed after induction by the isopropyl-beta-D-thiogalactopyranoside (IPTG) and was analyzed with SDS PAGE. RESULTS: The recombinant plasmid pET-SOD was obtained, and the recombinant protein was highly expressed in E. coli BL21(DE3). The activity of recombinant superoxide dismutase was 51,800 U per gram of wet bacteria. CONCLUSION: This study has provided a foundation for further studies and applications of the recombinant Mn-SOD. PMID- 15807267 TI - [Expression with TGFbeta1 in the patients with ulcerative colitis]. AB - OBJECTIVE: To elucidate the differences in expression of TGFbeta1 and TGFbeta1 mRNA between ulcerative colitis (UC), infectious colitis (IC) and normal control. METHODS: TGFbeta1 in colonic mucosa was detected by immunohistochemistry (IHC). TGFbeta1 mRNA was detected by hybridization in situ. RESULTS: There was no difference in detecting TGFbeta1 expression and TGFbeta1 mRNA expression in colonic mucosa between UC group and IC group (P>0.05), but the expression rates for the two groups were significantly higher than those for normal control (P<0.001). The expression of TGFbeta1 in colonic mucosa of UC group was noted to have a positive correlation with UC histological grade (r=0.462, P=0.002). CONCLUSION: Enhanced TGFbeta1 production in the colonic mucosa of UC patients can not inhibit proinflammatory cytokine production and hence can not get control of inflammation; this finding suggests the possible presence of TGFbeta1 signaling defects in the cases of UC. TGFbeta1 may serve as a disease activity marker of ulcerative colitis. PMID- 15807268 TI - [The clinical, pathological features and expression of Ki-67 and COX-2 in severe reflux esophagitis and Barrett's esophagus]. AB - OBJECTIVE: To study the clinical, pathological features of severe reflux esophagitis (SRE) and Barrett's esophagus (BE) and search for expression and significance of Ki-67 and COX-2 in their tissues. METHODS: Both SRE (n = 15) and BE (n = 25) cases were retrospectively analyzed for clinical manifestations, endoscopic findings and pathological features. The expression levels of Ki-67 and COX-2 of esophageal epithelium in the two groups were compared with that of normal esophageal (NE) epithelium (n = 10) by immunoperoxidase staining. RESULTS: It was found that the positivity and staining intensity of Ki-67 expression in SRE and BE were higher than those in normal esophageal epithelium (P<0.01), but there were no statistical differences between these two groups (P>0.05); COX-2 was selectively expressed in some BE epithelium, but not in SRE and normal esophageal epithelium. CONCLUSION: There were intensified Ki-67 expressions in the epithelium of SRE and BE, which could help us to evaluate the prognosis of gastroesophageal reflux disease (GERD) patients clinically by surveillance of Ki 67. COX-2 was selectively expressed in BE epithelium, so COX-2 inhibitors may have the potential for treatment of BE and for chemoprevention of its malignant change. PMID- 15807269 TI - [Study on the mechanism by which Rhubarb protects the mucosal barrier of intestine of mouse]. AB - OBJECTIVE: To observe the influence of Rhubarb on the the excretion of Type II PLA2 and lysozyme of small intestine of mouse. METHODS: Forty ICR mice were randomized to two groups. In the experiment group, the mice were gavaged with 0.3 ml 10% Rhubarb decoction every 8 hours; in the control group, the mice were given normal saline instead of Rhubarb decoction. After 24 hours, the mice were subjected to cervical dislocation, and their jejunum and ileum were taken out. The lumen of each resected intestine was rinsed with 100 g/L acetic acid, and the washed intestines from each mouse were cut into pieces 1-2 mm in length. Then the perfusate and homogenate were prepared, lyophilized, sealed, and stored at -20 degrees C. The Type II PLA2 activity and lysozyme were assayed respectively. RESULTS: The Type II PLA2 activities and lysozyme of homogenate in Rhubarb group were lower than those in Saline group (P<0.01). The type II PLA2 activities and lysozyme of perfusate in Rhubarb group were higher than those in Saline group (P<0.01). CONCLUSION: Rhubarb can stimulate the small intestine of mice to excrete Type II PLA2 and lysozyme, thus resulting in the increase of Type II PLA2 and lysozyme contents in the intestinal tract and enhancing the function of mucosal barrier of intestine. PMID- 15807270 TI - [Preventive effects of montelukast on the collagen expression of pulmonary arterioles in rats with chronic hypoxia]. AB - OBJECTIVE: To evaluate the preventive effects of montelukast on the collagen expression of pulmonary arterioles in chronic hypoxic rats. METHODS: Thirty male Wistar rats were randomly divided into three groups: control group, hypoxic group and montelukast preventive group. The animal model of pulmonary hypertension was established by exposing the rats to normabaric hypoxic conditions 8 hours q.d. for 3 weeks. The expression levels of collagen I and III in arterioles were observed by immunohistochemistry. RESULTS: The positive degree of collagen I in pulmonary arterioles of hypoxic group was higher than that of control group (1.51+/-0.09 vs 1.15+/-0.05, P<0.01), and the positive degree of collagen I in pulmonary arterioles of preventive group (1.19+/-0.06) was lower than that of hypoxic group (P<0.01). The differences of positive degree of collagen III in pulmonary arterioles were not significant among the three groups (P>0.05). CONCLUSION: Montelukast can reduce the hypoxia-induced deposition of collagen I in the pulmonary arterioles wall. PMID- 15807271 TI - [Anticarcinogenic effect of 20(R)-ginsenoside Rg3 on induced hepatocellular carcinoma in rats]. AB - OBJECTIVE: To explore the anticarcinogenic mechanism of 20(R)-ginsenoside Rg3 in induced liver tumor in SD rat. METHODS: Thirty-five SD rats with induced hepatocellular carcinoma were divided into a control group and 3 dosage groups according to the dosing levels of 20(R)-ginsenoside Rg3. The tumour volume was measured by MR imaging. The apoptotic rat and S-phase fraction and diploid of tumor cell were measured with flow cytometry. Protein expression of PCNA and TNF were evaluated with immunohistochemistry. RESULTS: There was significant difference in tumour volume between the high dosage group and the control group. The average apoptotic rates were 11.08+/-3.78, 13.57+/-3.34, 27.35+/-16.04 and the S-phase fractions were 23.98+/-9.44, 19.73+/-6.62, 14.09+/-3.48 in the low-, medium-, and high-dosage groups respectively. The apoptotic rate was significantly higher in the high-dosage group than in the medium-dosage group and low-dosage group. Before-after comparison showed that the anti-proliferative effects of 20(R)-ginsenoside Rg3 were significant in three treatment groups. The higher positive rats of protein expression with PCNA and TNF were significant difference in the high-dosage group compared to those in the low-dosage group. No significant difference between the medium-dosage group and the low-dosage group. CONCLUSION: 20(R)-ginsenoside Rg3 can noticeably inhibit the proliferation of tumor cells, and efficaciously induce the apoptosis and facilitate necrosis of the tumor cells, and there appears to be a dose dependent effect. PMID- 15807272 TI - [Effects of Radix Salviae Miltiorrhizae on Ca2+ in hepatic stellate cells]. AB - OBJECTIVE: To investigate the effects of Radix Salviae Miltiorrhizae (RSM) on intracellular free calcium in hepatic stellate cells (HSCs). METHODS: After the model of hepatic fibrosis was established in SD rats, RSM [20 ml/(kg x d)] was given via gastrogavage to the rats of treatment groups while the same volume of 0.9% NaCl was given to the rats of control groups twice a day for 6 consecutive days. Then the blood sample was drawn from the inferior vena cava, and the serum was extracted for pharmacological studies. After 24 h incubation with 10% drug serum, HSCs were loaded with Fluo-3/AM, a Ca2+ marker, and were observed with laser scanning confocal microscopy (LSCM). RESULTS: By comparison with controls, both RSM pharmacological serums decreased [Ca2+]i in HSCs significantly in the condition of using Ang II or not (P<0.05). CONCLUSION: RSM decreased [Ca2+]i in activated HSCs, which may be one of important ways to block liver fibrosis. PMID- 15807273 TI - [Effects of L-threonate on bone resorption by osteoclasts in vitro]. AB - OBJECTIVE: To clarify if calcium L-threonate and sodium L-threonate have inhibitory effects on the bone resorption of rabbit's osteoclasts in vitro. METHODS: This study contained a total of 16 culture groups, including one group as control and 5 groups treated by 5 drugs (calcium D-threonate, sodium L threonate, alendronate, 17beta-estradiol and calcium gluconate) each at the final concentrations of 10(-9) mol/L, 10(-7) mol/L, 10(-5) mol/L respectively. After 7 days, eight bone slices of every group were stained with toluidine blue and the areas of resorptive pits were analyzed under light microscope; the concentrations of C-telopeptide of type I collagen (CTx or Crosslaps) in culture supernatants were measured by ELISA. RESULTS: (1) The resorption area and the CTx concentration of the Calcium L-threonate groups were reduced significantly as compared with those of control and of Calcium gluconate groups respectively. The resorption area and CTx level of the Sodium L-threonate groups were significantly reduced when compared with those of the control, but the effects of Calcium gluconate groups were not so. (2) The reduction in the resorption area and CTx concentration of Calcium L-threonate group was more than that of Sodium L threonate group. (3) The reductive effect of the high concentration (10(-5)) group of Calcium L-threonate on the area and CTx level was corresponding to that of 17beta-estradiol at a concentration between 10(-7) and 10(-9). (4) The resorption area was related to the CTx concentration (r=0.876). (5) The CTX level was much more sensitive, precise and stable than the concentration. CONCLUSION: L threonate, especially calcium L-threonate could inhibit the bone resorption of osteoclasts in vitro, and its effect might be related to the radical of L threonic acid. The CTx concentration in culture supernatants might be an effective marker quantitatively reflecting the bone resorption by osteoclasts in vitro. PMID- 15807274 TI - [The effect of retinoic acid on induction of osteoporotic model rats and the possible mechanism]. AB - OBJECTIVE: To observe the effect of retinoic acid on induction of osteoporotic model in rats and analyze the mechanism therein involved. METHODS: SD rats were treated with retinoic acid 80 mg/(kg x d) by gastrogavage for 15 days to induce osteoporosis and were killed in batches at 0, 30 and 60 days after drug withdrawal. The levels of Ca, P, BGP, E2, IGF-1, AKP and TRAP in serum were assayed, the collagen and proteoglycan contents of bone and bone mineral density (BMD) were determined, and the morphological changes in cancellous and cortical bone and growth plate cartilage (GPC) of femurs from the experimental rats were observed. RESULTS: After 15 days of induction by retinoic acid, the serum E2 and BGP contents of rats were obviously decreased, the activities of AKP and TRAP were significantly increased, and the levels of BMD were lowered. The masses of spongy bone and cortical bone of femurs from the rats were decreased, and the number of chondrocytes in GPC was reduced. At 30 days, after drug withdrawal, the masses of spongy bone and cortical bone of femurs from the osteoporotic model rats still showed reduction; the activities of AKP in serum were lower than those at 15 days after drug redrawal, but were still higher than those of normal group rats; the chondrocytes in GPC were increased, the serum BGP and Ca contents were increased. At 60 days, after drug withdrawal, only the masses of femoral spongy bone of the osteoporotic model rats continuously showed obvious reduction, the other indices, including BGP, E2, AKP, TRAP and the masses of cortical bone, showed no significant difference between the two groups. CONCLUSION: The short term effect of retinoic acid on induction of rat's osteoporotic model was noticeable, but the long-term effect was not so good, and the bone loss of spongy bone existed longer and was more obvious than that of cortical bone. PMID- 15807275 TI - [Randomized controlled trial on haiguiyuyang capsule in the treatment of duodenal ulcer]. AB - OBJECTIVE: To assess the efficacy and safety of haiguiyuyang capsule in the treatment of duodenal ulcer (also diagnosed as weiwan pain and hanrecuoza syndrome according to the theory of TCM). METHODS: This is a multi-center clinical trial cooperatively conducted from May 2003 to March 2004 in accordance to the principle of informed consent, to the criteria for subject inclusion, exclusion, discontinuation, and to the relevant guidelines for evaluating the therapeutic effects of new TCM drugs. The design of double-blind, double-dummy and randomized controlled trial was adopted. 438 patients were randomized to the Test group (n = 330, treated with haiguiyuyang capsule) and to the Control group (n = 108, treated with ranitidine). The therapeutic course for both groups was 6 weeks. RESULTS: Regarding the efficacy in treating duodenal ulcer, in the Test group, the Marked Efficacy Rate was 66.37% and the Total Efficacy Rate was 82.13%; in the Control group, the Marked Efficacy Rate was 68.61% and the Total Efficacy Rate was 93.34%; there was no significant difference between the two groups (P>0.05). Regarding the efficacy in treating the syndrome diagnosed by TCM, in the Test group, the Marked Efficacy Rate was 70.31% and the Total Efficacy Rate was 93.34%; in the control group, the rates were 71.29% and 91.66% respectively; there was no significant difference between the two groups (P>0.05). Besides, regarding the abatement of distention and fullness of stomach after treatment, the haiguiyuyang capsule was better than ranitidine (P<0.05). No adverse side-effect was observed. CONCLUSION: The efficacy of haiguiyuyang capsule in treating duodenal ulcer is similar to that of ranitidine. No obvious adverse effect of it was observed in this trial. PMID- 15807276 TI - [The effect of ultraviolet B on interleukin-8 secretion in human keratinocyte cell line]. AB - OBJECTIVE: To investigate the effect of ultraviolet B (UVB) on Interleukin-8 secretion in human keratinocyte cell line. METHODS: The concentration of IL-8 was detected by ELISA kit 24 h after human keratinocytes (HKC) were irradiated by different doses of UVB, and the level of IL-8 was also determined at different times after the same dose of UVB irradiation. RESULTS: The secretion of IL-8 was increased after the HKC were irradiated by UVB, the effect was dose-dependent when UVB ranged from 10 to 40 mJ/cm2, and there was statistically significant difference between the IL-8 level of UVB groups (20-70 mJ/cm2) and the control (0 mJ/cm2) (P<0.01). The level of IL-8 was increased 1 h after the irradiation of 30 mJ/cm2 UVB, and it reached the peak at 12 h. There was statistically significant difference between the IL-8 levels detected at different times (3 h, 6 h, 12 h, 24 h after irradiation) and that at 0 h (P<0.01). CONCLUSION: UVB increase the secretion of IL-8, and the effect is dose-dependent to some extent. PMID- 15807277 TI - [Expression and significance of mRNA for MIP-1alpha in cerebral tissue of newborn rat with hypoxic-ischemic brain damage]. AB - OBJECTIVE: To observe the expression of mRNA for the MIP-1alpha in cerebral tissue after hypoxic-ischemic brain damage (HIBD) in newborn rat. METHODS: The dynamic changes of expression of MIP-1alpha mRNA were studied using 7 d-old SD rat hypoxic-ischemic brain damage model and quantitative, real-time, fluorogenic PCR (TaqMan) assay. RESULTS: Peaking transcription was shown at 6 hours after HIBD when MIP-1alpha mRNA level in the HI cerebral hemisphere was almost 10 times compared with that of control group. The mRNA level of 12 and 24 hours after HIBD was also greater than that of control group. CONCLUSION: This appearance suggests that, as an inflammatory mediator, the MIP-1alpha transcription after HIBD may play important roles in the evolvement of brain damage. PMID- 15807278 TI - [The expression of Fas after fluid percussion brain injury in rats]. AB - OBJECTIVE: To inquire into the pathologic diagnosis and the dating of injuries of light closed encephalon injury. METHODS: Wistar rats were hurt by fluid percussion, and were killed respectively at 15 min, 30 min, 1 h, 3 h, 6 h, 12 h, 1 d, 4 d, 7 d, and 14 d after injury. The expression of Fas in the cerebral cortex, thalamus, and hippocampi was detected by immunohistochemistry and the results were assessed by image analysis system. RESULTS: The results showed that the expression of Fas could be detected in thirty minutes after injury, and then it increased significantly in three hours, reached apex in twelve hours after injury, decreased gradually in four days after injury, and returned to normal in 14 days after injury. CONCLUSION: This research demonstrated that Fas-mediated apoptosis appeared not only around brain trauma but also in the brain tissue far away from the traumatic area. It indicted that Fas expression is a useful target for diagnosis of early brain injury and the regularity of Fas expression could be used as one of indications in deducing the time of brain injury. PMID- 15807279 TI - [The protective efficacy of rabbit endogenous nitric oxide against acute rabbit lung ischemia-reperfusion injury and its mechanism]. AB - OBJECTIVE: To investigate the protective efficacy of rabbit endogenous nitric oxide (NO) against acute rabbit lung injury associated with ischemia-reperfusion and explore the possible mechanism. METHODS: The rabbit lung ischemia-reperfusion (LIR) model was established; thirty-two adult New Zealand white rabbits were randomly divided into four groups. The rabbits of control group underwent sham operation. In group LIR, the rabbits' left lung hili were clamped for 60 minutes and then released. In group LIR+L-Arg (L-arginine), the rabbits were operated upon as those in group LIR, but L-Arg (200 mg/kg) was infused into blood circulation as substrate for NO generation before removal of the clip. In group LIR+L-NNA (L-ng-nitro-Arginine), the rabbits passed through the same operation as in group LIR, but L-NNA (10 mg/kg) was infused into circulation as an inhibitor against NO generation before reperfusion. After reperfusion for 60 minutes, the lung tissues were harvested for histological examination, and the wet to dry ratio of lung tissue weight (W/D), myeloperoxidase (MPO) activity, malondialdehyde (MDA) content as well as the ratio of nitrate/nitrite (N/N) were measured respectively. RESULTS: The group LIR had greater lung tissue W/D, higher MPO activity and MDA content, lower ratio of N/N, and serious pulmonary edema as compared with group ShO (P<0.01). But in group LIR+L-Arg, the degree of pulmonary edema was alleviated, the MPO activity and MDA content were decreased, and the ratio of N/N increased; there was statistically significant difference between group LIR and group LIR+Arg in respect to the above indices (P<0.01). However, in group LIR+L-NNA, the pulmonary edema was even more severe, the MPO activity and MDA content were significantly higher those that in group LIR or group LIR+L-Arg (P<0.01). CONCLUSION: The endogenous release of pulmonary NO can attenuate the acute lung injury associated with LIR, and the mechanisms may involve the protective efficacy conferred by endogenous NO against accumulation of neutrophil in lung, against pulmonary microvascular permeability, and against the oxygen free radical injury to lung. PMID- 15807280 TI - [Comparative study on the genotoxicity of gasoline-fueled vehicle exhaust and methanol-fueled vehicle exhaust]. AB - OBJECTIVE: Comparing the genotoxicity of gasoline-fueled vehicle exhaust (gasoline exhaust for short) with that of methanol-fueled vehicle exhaust (methanol exhaust for short) so as to provide a scientific basis for replacement of gasoline by methanol as fuel in vehicle. METHODS: The MTT method was used first to evaluate the cytotoxicity of the two kinds of vehicle exhausts, and the concentration that had no obvious cytotoxicity would be used as the highest dose in the experiments. The A549 cells micronucleus test and single cell gel electrophoresis (SCGE) assay (comet assay) in vitro were applied to compare the genotoxicity of gasoline exhaust and methanol exhaust. RESULTS: The MTT results showed that the gasoline-fueled vehicle exhaust exerted stronger cytotoxicity to A549 cells in both 2 h and 24 h exposure times, compared with the methanol exhaust. In A549 cells micronucleus test in vitro, at doses 0.025, 0.05, 0.1 and 0.2 L/ml, the micronucleated cell (MNC) rates were 2.65%, 4.35%, 4.95% and 5.85% respectively, which were higher than those (1.30% and 1.35%) of controls (P<0.01). For the methanol-fueled vehicle exhaust, there was no significant difference in the rate of A549 cells micronucleus between the test groups and control group. In the comet assay, gasoline-fueled vehicle exhaust could induce A549 cells DNA damage. The rate of caudate cell and the length of DNA migration increased with the escalation of dosing level. However, the methanol-fueled vehicle exhaust did not show any DNA damage to A549 cells. CONCLUSION: The results demanstrate that gasoline-fueled vehicle exhaust can induce DNA and chromosome damage, it has a distinct genotoxicity, whereas the methanol-fueled vehicle exhaust does not show any potential genotoxicity in both tests. PMID- 15807281 TI - [Proton spectroscopy findings of meningioma]. AB - OBJECTIVE: To evaluate the characteristics of 1HMRSI and its diagnostic value on meningioma. METHODS: The findings of conventional MRI and of 1HMRSI from 11 cases of clinically proven meningiomas and from 12 normal volunteers were comparatively analyzed. The imaging sequences included T1-weighted imaging and T2-weighted imaging; additionally T1-weighted imaging with injection of the contrast agent of Gd-DTPA was performed in all meningioma cases. Two-dimensional phase encoding of proton magnetic resonance spectroscopic imaging (2D 1HMRSI) were obtained in all cases. RESULTS: On conventional pre-contrast MRI, the signals in 8 meningioma cases showed medium or faintly low intensities and in 3 cases showed mixed intensities on T1WI. On T2WI, the signals in 7 cases showed medium or faintly high intensities and in 4 cases showed mixed intensities. After injection of Gd DTPA, the solid portion of tumors exhibited obvious enhancement in all meningioma cases. "Dural trail sign" was revealed in 7 cases. Peri-tumor edema was evident in 11 meningioma cases. Compared with control group, 1HMRSI of all meningiomas revealed different spectral peaks, including absent acetylaspartate (NAA), prominent choline (Cho), and reduced phosphocreatine (Pcr). The ratios of Cho/Pcr obviously increased. The peak of lactate (Lac) was visualized in 2 meningioma cases. CONCLUSION: Most cases of meningioma can be diagnosed with conventional magnetic resonance imaging (MRI). 1HMRSI can yield more informative findings about meningioma via the observed metabolic materials changes in tumor cells. So conventional MRI is the most important technology for diagnosing meningioma and 1HMRS combined with MRI can improve the diagnostic accuracy. 1HMRSI can be an important supplemental means in the diagnosis of meningiomas. PMID- 15807282 TI - [CT differentiation of renal angiomyolipoma and renal cell carcinoma]. AB - OBJECTIVE: To improve the diagnosis of atypical renal angiomyolipoma (RAML) by use of CT to differentiate RAML from renal cell carcinoma (RCC). METHODS: We retrospectively analyzed the CT manifestations with dual-phase enhanced contrast study in 36 cases of RAML (with 57 lesions) and 46 cases of RCC. 25 lesions of RAML and all RCC were histologically confirmed. Maximum likehood estimator was employed to discriminate all the lesions according to a series of indexes with statistical meanings. RESULTS: Nine indexes were found to be significant and evident for separating RAML from RCC, namely "single or multiple", border, lobulation, protrusion rate, interface with cortex, angle with cortex, "levering cortex-up" sign, characters of blood vessel in lesion and calcification. The accuracy rates of discrimination analysis for RAML and RCC were 93.0% and 89.1%, separately. CONCLUSION: The importance of 4 indexes, including protrusion rate, angle with cortex, "levering-cortex-up" sign and the traits of blood vessel, should be highlighted in the differentiation of RAML and RCC. PMID- 15807283 TI - [The regularity of changes on the CT images of accessory nasal cavity in patients with nasopharyngeal carcinoma after radiotherapy]. AB - OBJECTIVE: To analyze the CT findings of nasal sinusitis in patients with nasopharyngeal carcinoma post-radiotherapy and detect the regularity of their change in different periods. METHODS: This is a study on 145 cases of nasopharygeal carcinoma, including 75 cases treated by gamma-ray and 70 cases by X-ray. CT examinations on the patients were carried out pre-radiotherapy and post radiotherapy in different periods. RESULTS: The radioreactions in maxillary sinus and sphenoidal sinus might be classified as patch form, crescent form, ring form and polyform. The unit of radioreaction in ethmoidal sinus was ethmoidal cell. Several ethmoidal sinus cells were fused to patch in severe cases. The radioreactions in ethmoidal sinus, maxillary sinus and sphenoidal sinus were seen within three months, after three months and after one year respectively. And nasal sinus radioreactions of most cases were not absorbed spontaneously after one year. When the two kinds of radiotherapies were equal in dosing level, there was no obvious difference between them in respect to the radioreaction of accessory sinus, but when the dosing level was above 3182 GBq, the radioreaction of accessory sinus was remarkably increased, compared with those caused by lower dose radiotherapies. CONCLUSION: CT plays an important role in yielding helpful information for the diagnosis and treatment of the nasal sinusitis of nasopharyngeal carcinoma post-radiotherpy. The patients' life quality can be improved. PMID- 15807284 TI - [Fracture resistance of the all-ceramic posts in post-and-core system]. AB - OBJECTIVE: To make a comparative study on the fracture resistance of the all ceramic posts in post-and-core system where the ceramic core was combined with the post by direct sintering and by two other methods. METHODS: A total of 15 all ceramic posts fabricated by alumina-zirconia nano-composite infiltrated ceramics were randomly divided into three groups. They were given the following treatments: Group A: forming and combining the ceramic core with the post by direct sintering; group B: combining the ceramic core with the post by use of 3M RelyX for adhesion; group C: forming composite resin core onto the post. Each specimen was placed into a special jig and subjected to a load perpendicular to the post axis on INSTRON 4302 universal testing instrument until failure. ANOVA was used for statistical analysis. RESULTS: There was no statistically significant difference between the groups (P>0.05). CONCLUSION: The findings indicate that the strength of the post remains uninfluenced during the combination of the core with the post by direct sintering. PMID- 15807285 TI - [Comparison of efficacy between nateglinide and repaglinide in treating type 2 diabetes: a randomized controlled double-blind clinical trial]. AB - OBJECTIVE: To compare the efficacy of nateglinide with repaglinide in the treatment of type 2 diabetes mellitus. METHODS: Forty-six type 2 diabetic patients were randomly treated with repaglinide (group A, 1.0 mg tid, n=23) or nateglinide (group B, 90.0 mg tid, n=23). The trial consisted of a 4-week equilibrated period followed by 12 weeks of treatment course. RESULTS: In group A, the fasting blood glucose (FBG) and 30-, 60-, 120- min postprandial blood glucose (PBG), as well as hemoglobin A1c were decreased significantly (P<0.05). In group B, the 60-min and 120-min PBG decreased remarkably (P<0.05), but FBG, 30 min PBG and A1c decreased with no statistical significance (P>0.05). After 12 weeks treatment, the 30-, 60-, 120-min postprandial insulin level, area under the curve of insulin and C peptide (0 to 120 min) increased in both groups (P<0.05). No significant difference was found between the effects of repaglinide and nateglinide on early phase insulin secretion. CONCLUSION: The glucose lowering effect of repaglinide at a dosing level of 1.0 mg tid was better than that of nateglinide 90 mg tid on fasting blood glucose and A1c during 12 weeks treatment period, yet the insulinotropic effects of the two drugs were similar. PMID- 15807286 TI - [Relationship between the adult attachment styles of social anxiety disorder (SAD) and its cognitive mode and behavior mode]. AB - OBJECTIVE: The adult interpersonal relationship is related to individual's early attachment experience. Insecure attachment relationship may lead to dysfunction of interpersonal relationship. This study was carried out to disclose the relationship of SAD-based adult attachment styles with cognitive mode and personality traits. METHODS: A comparison was made between the SAD group and normal control (NC) group by using the psychohygiene measures: Adult attachment scale (AAS), Social avoidance and distress scale (SADS), Fear of negative evaluation scale (FNE), Texas social behaviour inventory (TSBI), and Status trait anxiety inventory (STAI). Statistically, T test, ANOVA, and Scheffe multiple comparison were used. RESULTS: Chi-square test revealed a significant adult attachment-style difference between SAD group and NC group; T test revealed significant FNE, SADS, TSBI and STAI differences between SAD group and NC group. One-way ANOVA revealed significant difference of FNE, SADS, TSBI and STA on the adult attachment styles within SAD group. CONCLUSION: The internal working models of fearful and preoccupied styles in adult attachment are the pathopsychological basis of SAD. PMID- 15807287 TI - [Secular trends of hypospadias in Chinese perinatals]. AB - OBJECTIVE: To investigate the secular trends in the prevalence rate of hypospadias in Chinese perinatals and explore its possible influencing factors. METHODS: Data were taken from Chinese Birth Defects Monitoring Network, a hospital-based congenital anomalies registry system. All live or still births at or beyond 28 weeks of gestation were assessed within 7 days after delivery in the period from 1987 to 2001. RESULTS: 1999 cases of hypospadias were identified among 3,769,464 male births, the crude prevalence rate being 5.30 per 10,000 births. And an increasing secular trend was observed both in urban and rural areas. In recent years, an increase in rate of hypospadias was found in coastal provinces and inland provinces, but not in remote border provinces. Faster increase was seen in urban areas and coastal areas, compared with that in rural areas and inland areas respectively. CONCLUSION: The prevalence rates of hypospadias in Chinese perinatals from 1987 to 2001 showed an upward tendency. PMID- 15807288 TI - [Purification of human serum apolipoprotein A I and A II by middle-pressure liquid chromatography]. AB - OBJECTIVE: This study is made for the purification of human HDL apolipoproteins by middle-pressure liquid chromatography (MPLC). METHODS: Human HDL was isolated by one step density ultracentrifugation. Delipided human HDL was separated by Sephacryl S-200 molecular sieve chromatography. Sephadex G-75 molecular sieve chromatography was used to separate apoA I and apoA II. RESULTS: Purified apoA I and apoA II were obtained and SDS-PAGE and immunodiffusion test indicated that the proteins are the same as those theoretically predicted. CONCLUSION: We have established a purification procedure for human HDL apolipoproteins with high efficiency and simplicity by MPLC. It could serve as a base for preclinical and clinical trials of HDL apolipoproteins. PMID- 15807289 TI - [Purification and identification of recombinant varicella-zoster virus glycoprotein E]. AB - OBJECTIVE: To purify and identify recombinant Varicella-Zoster Virus Glycoprotein E. METHODS: The recombinant plasmid pGEX-VZVgE was induced by isopropyl-beta-D thiogalactoside (IPTG), the fusion protein was purified with affinity chromatography column; then the purified fusion protein was cleaved by thrombin, and the product's antigenicity was examined by Western blot. RESULTS: The product of pGEX-VZVgE induced by IPTG was separated from the mixture proteins by the affinity chromatography column, the expressed fusion protein's relative molecular mass was about 98 x 10(3). After cleavage, the obtained VZV Glycoprotein E's relative molecular mass was about 72 x 10(3); the purified fusion protein and VZV Glycoprotein E were single band by SDS-PAGE. The available antigenicity of Glycoprotein E was confirmed by Western blot. CONCLUSION: Purification of VZV Glycoprotein E with affinity chromatography is an effective method. It provides a foreground for studies on the application of VZV gE. PMID- 15807290 TI - [Current and perspective treatment of chronic lymphocytic leukemia]. AB - Chronic lymphocytic leukemia represents the most frequent hematological malignancy in the western population. Important role in its pathogenesis has the inhibition of apoptosis, disturbances in differentiation of B-lymphocytes, slightly increased proliferation and induction of angiogenesis. Current treatment options (chemotherapy and immunotherapy) can improve the quality of life and prolong disease-free survival, but the overall survival is not significantly influenced. The only curative treatment alternative is the allogeneic hematopoietic stem cell transplantation. However, this treatment is accompanied by the risk of peritransplantation mortality and its use is limited for younger patients. Number of new compounds is clinically tested. New drugs can influence intracellular events such as induction of apoptosis or inhibition of the transduction cascade. Review summarises treatment alternatives and presents an algorithm for their clinical application. PMID- 15807291 TI - [Endocannabinoids--the new option in the treatment of metabolic syndrome and in smoking cessation]. AB - Development of the metabolic syndrome results from the interaction of genetic and environmental factors. Metabolic syndrome together with smoking represents risk factors for the development of cardiovascular complications. They may result from the hyperstimulation of the endocannabinoid system. The CB1 receptor has been assumed to play an important role in the endocannabionoid system. It is abundantly expressed in the brain, and in other parts of human body such as in the fat tissue. Rimonabant is a selective blocker of cannabinoid-1 (CB1) receptors and participates in the regulation of impaired endocannabinoid system. In the overweight humans, it stimulates sustained reduction of the body weight, girth size and it improves lipid and glucose metabolism. Rimonabant also reduces nicotine self-administration and may be effective not only as an aid for smoking cessation but also in the prevention of body weight increase related to the smoking cessation as it was documented in Rio-Lipids and Stratus-us studies. PMID- 15807292 TI - [Serum tumor markers at exocrine adenocarcinoma of pancreas]. AB - Pancreatic cancer represents a group of cancers with the worst prognosis. There exists only one method of successful therapy--radical surgery, i.e. the complete resection of the tumor. However, only 5% of cases are diagnosed in the early stage. More than 60% are recognised in the period of advanced disease. For those patients only palliative treatment can be used. Only early diagnosis can ensure better therapy results. Possibility of early diagnosis based on imaging and endoscope methods is at present very limited due to the anatomical location of pancreas. Serum tumor markers assessment might represent one of the possibilities of early detection of the pancreatic cancer. Review summarizes current knowledge on the use of tumor markers for diagnosis, prognosis and therapy monitoring of pancreatic exocrine adenocarcinomas with respect to the clinical use at the daily routine practice. PMID- 15807293 TI - [Promyelocytic leukaemia protein and defect in transforming growth factor-beta signal pathway in acute promyelocytic leukaemia]. AB - Chromosome translocations are detected in 50-70% of human leukaemia. The promyelocytic leukaemia (PML) gene is involved in the t(15;17) chromosomal translocation of acute promyelocytic leukaemia (APL). PML gene encodes a protein, which was shown to be concentrated in PML-nuclear bodies. Histone acetyltransferases and deacetylases, and chromatin-modifying proteins are accumulated in complexes with PML protein in these nuclear bodies giving the evidence of their role in transcription regulation. Physical interactions of PML protein with transcription factors, co-activators and co-repressors of transcription correspond with the role of PML in transcription regulation. PML plays an important role in apoptosis, proliferation and senescence of cells. PML gene is a tumour-suppressor gene and a product of its expression acts as a potent cell growth suppressor. All these activities of PML protein are ascribed to its nuclear functions. Cytoplasmic form of PML (cPML) is also very important and it is critical for transforming growth factor-beta (TGF-beta) signalling. Cytoplasmic PML interacts with two TGF-beta receptors (TbetaBRI and TbetaRII) and acts as a bridging factor between protein called Smad anchor of receptor activation (SARA) and Smad proteins and it plays a role in the transport of whole complex into the early endosomes in TGF-beta signalling. The loss of functional cPML induces not only APL but it might influence behaviour of cancer cells and their resistance to TGF-beta. PMID- 15807294 TI - [Epidemiological aspects of the female urinary incontinence]. AB - Urinary incontinence is defined as an involuntary loss of urine, which makes social and hygienic problem. It is a symptom with different causes. According to the typical clinical manifestation it is classified as stress, urge, reflex and paradox urinary incontinence. Loss of small amount of urine related to the increase of intraabdominal pressure (during coughing, sneezing or running) is characteristic for stress urinary incontinence. Sudden and uncontrollable voiding with loss of greater amount of urine is typical for urge incontinence. Reflex incontinence means that urinary bladder is emptying without voiding. Paradox incontinence is caused by an acquired smooth muscle weakness of the bladder and it manifests with incomplete emptying and with growing residual urine. Prevalence of urinary incontinence increases with age. Significant increase of female urinary incontinence symptoms is found in fifth and sixth decade. Urinary incontinence in young women is more a dynamic than a permanent symptom but the postmenopausal incontinence obviously does not disappear spontaneously. Urge and mixed incontinence are less frequent than stress symptomatology (between 10 and 15%). According to the prevalence studies only 1,5 to 6% of incontinent women are looking for a medical help. Because the urge symptoms are more limiting, the patients with urge incontinence are searching treatment possibilities more often than those with stress incontinence. PMID- 15807295 TI - [Diagnostic algorithms in female urinary incontinence]. AB - Urinary incontinence is frequent problem with significantly negative impact on the quality of life. The correct plan of examination leads to the proposal of the appropriate treatment. The history, correct judgment of symptoms of incontinence and detailed gynaecological examination are important at the beginning of the diagnostic process. Negative urine cultivation is necessary condition for the invasive investigation of the urinary tract. Special questionnaires, micturition diary and clinical tests belong to the physical examination. The pad-weight test is based on the measurement of weight gain of absorbent pads during a test period under standardised conditions. Q-tip test (cotton swab test) measures the degree of mobility of urethra and bladder neck. Most of the clinical tests can be replaced by the ultrasound examination (introital or perineal approach), which helps to appreciate the vigour of urethra, bladder neck and vagina during the resting period and at stress. Urodynamic examination records pressure changes during the bladder filling, pressure gradients in urethra and the velocity of the urinary flow. It should be carried out when antiincontinence surgery is planned or when conservative therapy failed. Urodynamic examination helps to distinguish between stress and urge type of incontinence. Urethrocystoscopy is indicated for the diagnosis of the recurrent urinary infections, urge incontinence when structural pathologies of the urinary bladder are suspected, iatrogenic incontinence or postoperative complications. PMID- 15807296 TI - [Therapy of acute myocardial infarction in unspecialised medical centres- evaluation of diagnostic and therapeutic practice]. AB - BACKGROUND: No general register of patients with acute coronary syndrome is available in the Czech Republic. That is why the everyday clinical practice remains indeterminate. The aim of the study was to compare patients' characteristics, diagnostic and therapeutic procedures in patients with acute myocardial infarction admitted to three municipal hospitals (Caslav, Kutna Hora, Znojmo) and to assess the correspondence of the local clinical practice with the current guidelines. METHODS AND RESULTS: A total of 564 patients (300 men, 264 women) mean age 71 +/- 11.9 years) admitted for acute myocardial infarction was included into the study. No significant difference was found in the demographic data among the hospitals. Higher proportion of smokers was observed in the hospital of Kutna Hora (p < 0.01). In the studied sample, the proportion of myocardial infarctions without ST elevation was larger than that without it (311 pts. - 73%). In the subgroup of patients with myocardial infarction and ST elevation 67% received some form of reperfusion therapy (primary angioplasty in 63 % and fibrinolytic therapy in 4%). Most patients received aspirin and heparin (78% a 82% respectively) within the first 24 hours; the treatment with beta blockers was less common (34%). The most frequent in-hospital complication was the heart failure (23%). In-hospital mortality for all patients was 15%. At the discharge from hospital, patients received flowing treatment: aspirin (76%), beta blockers (63%), ACE inhibitors (58%), statins (54%). A significant difference was found only in the use of clopidogrel and ticlopidin (p < 0.001). CONCLUSIONS: This study reveals only minor differences in the management of patients among hospitals and demonstrates some differences between existing guidelines and current clinical practice. The vast majority of patients with acute myocardial infarction with ST elevation was treated by primary angioplasty. PMID- 15807297 TI - [The use of auto mode switching in patients with sick sinus syndrome]. AB - BACKGROUND: At present the dual chamber pacing, originally developed for patients with AV blockades, is widely used also for patients with Sick sinus syndrome (tachycardic-bradycardic type). But these patients often cause therapeutical problems to their physicians. In these cases either antiarrhythmic therapy is necessary to prevent recurrent supraventricular tachycardias (which are cause of rapid ventricular pacing) or in the case of failure of AA therapy the pacing mode has to be changed to DDI/R, which excludes physiological VAT pacing. The Auto Mode Switching (AMS) function ensures adequate ventricular pacing rate in the time of SV arrhythmias. METHODS AND RESULTS: Effects of dual chamber pacemakers equipped with AMS were studied in a group of patients with paroxysmal atrial fibrilation and/or atrial flutter. CONCLUSIONS: Therapy brings effects in lower of expenses due to less frequent visits at the physician, lower rate of rehospitalizations and lower need for powerful AA therapy. PMID- 15807298 TI - [Molecular genetic study of causes of the Prader-Willi and Angelman syndrome]. AB - BACKGROUND: Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are distinctive diseases with severe impairment of psychomotoric development and behaviour. Both syndromes are caused by the loss of paternal (PWS) or maternal (AS) gene expression of chromosomal region 15q11-13. The work reveals the various causes of this loss. The choice of the most suitable method for screening of the genome mutations in the patients suspected of PWS and AS is another purpose of the work. METHODS AND RESULTS: The methyl specific analysis (MS PCR) in locus SNRPN, short tandem repeat (STR) analysis and fluorescent in situ hybridization (FISH) were used. In the group of 55 patients tested for PWS and AS only maternal allele was present in 11 patients and only paternal allele was present in 1 patient in the locus SNRPN: 10 microdeletions 15q11-13, 1 UPD(15)mat and 1 UPD(15)pat. CONCLUSIONS: MS PCR seems to be the most profitable method for the first step of selection of PWS patients. In positive cases is inevitable to use also additional tests of molecular diagnosis to distinguish the particular mechanism leading to the disorders. In AS patients is also MSPCR recommended as the first step although it is necessary to exclude mutation in UBE3A gene in case of MS PCR negativity. PMID- 15807299 TI - [Long-term follow up of renal function after orthotopic liver transplantation]. AB - BACKGROUND: Renal dysfunction in patients after the orthotopic liver transplantation (OLT) is frequent and it significantly contributes to the morbidity and mortality. The aim of our work was to assess the level of glomerular function (GFR) within the first to fifth year after OLT. METHODS AND RESULTS: Serum creatinine concentration (Skr), creatinine clearance (Ckr) and predicted value of creatinine clearance using the Cockcrofta a Gaulta formula (CG) were assessed in 75 patients. Normal values of the given parameters (Skr <110 umol/l, Ckr > or = 1.3 ml/s/1.73 m2) were found only in 16% of all patients. Significant decrease of GFR (Ckr < 0.5 ml/s/1.73 m2) was found in 24% of cases, acute renal failure, which required transitory haemodialysis developed in 4% of patients. In 60% of patients various degree of GFR decrease was found without the necessity of haemodialysis. CONCLUSIONS: Level of renal functions was not significantly related to the blood pressure or serum lipids concentration. An important factor appeared to be the level of renal function before OLT. Because the level of renal function after OLT can significantly influence the post transplantation development, regular follow up of GFR is recommended. PMID- 15807300 TI - [Organ changes induced by ergot derivative dopamine agonist drugs: time to change treatment guidelines in Parkinson's disease?]. AB - Ergot derivative dopamine agonists, e.g. pergolide, bromocriptine, dihydroergocriptine used in treatment of Parkinson's disease can cause pleural, pericardial, retroperitoneal and valvular fibrotic changes. Case No 1: A 56-year old woman with PD was treated with pergolide 3mg/24h since July 2002. In June 2003, edema of lower extremities was first noticed and echocardiography found a minor mitral regurgitation without any morphological changes of the valve. In January 2004, left- sided cardiac failure rapidly developed and echocardiography revealed multivalvular insufficiency with predominating severe mitral regurgitation. Mitral valve replacement was performed and pergolide was changed to ropinirole. Until now, neither cardiac functions nor motor status are sufficiently compensated. Case No 2: A 66-year-old-man with PD since 1996 was treated with pergolide 3 mg/day since 1999. In the beginning of 2004, leg edema appeared. On examination, bilateral hydronephrosis with ureteric strictures and incipient renal insufficiency was found. Bilateral ureteroplasty was performed and the histology showed periureteric fibrosis. Treatment with steroids was initiated and pergolide was changed to pramipexole. Despite the treatment, the fibrosis progressed, requiring ureteral stenting. Based on the literature review and on our own experience, we propose following guidelines to minimize the risk of complications: A. Not to use EAD as the first-line dopamine agonists. B. Regularly follow all patients treated with EAD, especially monitor the majorsymptoms: dyspnea, cough, fatigue, leg edema (also asymmetric), symptoms of urinary outflow obstruction, cardiac insufficiency, chest pain, heart murmur. An elevated ESR, C-reactive protein or anemia support the diagnosis. C. All symptomatic patients should undergo workup for serosal fibrosis (according to type of complication): chest X-ray or CT scan, spirometry, renal functions, renal ultrasound, CT of retroperitoneum. D. Before the introduction of EAD therapy, examine the renal functions, perform chest X-ray and echocardiography. Screening echocardiography should be performed in 3-6 months and subsequently in every 6-12 months. PMID- 15807301 TI - [Contribution to neglecting of power analysis in experimental medicine studies]. AB - The work provides basic information about what is being understood under the term "statistical analysis of power of test" from the viewpoint of correct use of mathematical statistics in medicine. The detail rules of statistical decision making are described. Furthermore, the work contains an illustration of power analysis of test application when using unpaired t-test. PMID- 15807302 TI - [Psychiatry is more than therapy]. AB - Presentation describes the relation between psychiatry and society and their mutual interplay. The special status of psychiatry among the medical disciplines is characterized. Indications and methods of restriction in psychiatry are given. Beside its curative and regulative function, psychiatry has to protect both the patients and the society. The means of restriction are presented (hospitalisation without the patient's consent, locked departments, physical and pharmacological restrictions, protective psychiatric treatment and other). Problems of medical juristic dilemma in psychiatric praxis are discussed. Official and unofficial methods of the regulation of patient's behaviour are described together with the necessity to develop and cultivate them. PMID- 15807303 TI - [Pharmacogenetics from the aspect of the Coordinating Group for Pharmacogenetics of OECD]. PMID- 15807304 TI - [Municipal Chief Health Counselor Professor MUDr. Bohdan F. Hejduk--incorrectly forgotten Czech surgeon]. PMID- 15807305 TI - Gastrointestinal stromal tumor (GIST) in southern Taiwan: a clinicopathologic study of 93 resected cases. AB - The purpose of this study was to determine the clinicopathologic features of gastrointestinal stromal tumor (GIST) in southern Taiwan. The pathology files from a medical center in southern Taiwan (1993 to 2003) were searched for primary mesenchymal tumors of the gastrointestinal tract. Hematoxylin/eosin sections and history were reviewed, and immunohistochemistry was performed using anti-CD 117, CD34, smooth muscle actin (SMA), and S-100 protein. Only primary resected GISTs were included in this study. Univariate and multivariate analyses were carried out using the T-test to evaluate the significance of primary tumor size and mitotic activity for the prediction of recurrence and metastasis. A total of 121 surgically resected primary mesenchymal tumors were identified, and 93 of these were GISTs. These 93 patients showed a slight female predominance (male: female = 1:1.2). The clinical presentations were variable and site-dependent. The most common tumor locations were the stomach (57%) and the small intestine (39%). Microscopically, 88 tumors (95%) were composed of spindle cells, the remaining five (5%) consisted of mixed epithelioid and spindle cells. No pure epithelioid type GIST was found. In addition to CD117, 66 cases (71%) were positive for CD34, 23 cases (25%) were positive for SMA, and 19 cases (21%) were positive for S-100. In a mean follow-up time of 27.3 months (median: 26 months), 19 cases (20.6%) were clinically malignant and mainly manifested as liver metastases (seven cases, 37% of malignant GISTs). Univariate analysis revealed that both primary tumor size and mitotic activity were significantly increased in the group affected by recurrence and/or metastasis (p = 0.001 and 0.035, respectively). Compared to GISTs in the western countries, those in southern Taiwan are characterized by a slight female predominance, a relatively higher frequency of small intestinal localization, a higher rate of S-100 protein expression, and a less aggressive behavior. Tumor size and mitotic activity were useful predictors of malignancy. PMID- 15807306 TI - Incidence and distribution of hybrid goblet cells in complete type intestinal metaplasia of the stomach. AB - Previous reports suggest that hybrid goblet cells (HGCs) sharing both gastric and intestinal mucin phenotypes are rarely observed in complete intestinal metaplasia (cIM) of the stomach. However, we have made a different observation. Thus, we compared the incidence and distribution of HGCs within the tubules of gastric cIM and the duodenum in order to define the significance of HGCs. Fifteen antral sections and 16 fundic sections from tissue with cIM and gastric cancer, as well as 19 sections from duodenal tissue with cancer of the Papilla of Vater, were stained for human gastric mucin (HGM), Con A, MUC2, CD10, and Ki-67. Multivariate analysis showed that antral location, a distance of 5mm or less from the tumor margin, and the presence of underlying pyloric glands were significant predictive factors for tubules containing >50% HGCs as part of their goblet cell population. The incidence of tubules with HGCs differed significantly in tissue samples from the antrum, body and duodenum. HGCs did not stain for Ki-67 and were not surrounded by gastric foveolar-type epithelium within the tubules of cIM foci. These findings indicate that alterations in the proportion of HGCs may occur under some circumstances, and that HGCs are not precursors to gastric foveolar type cells in the stomach and duodenum. PMID- 15807307 TI - Immunohistochemical expression of DNA mismatch repair (MMR) system proteins (hMLH1, hMSH2) in cervical preinvasive and invasive lesions. AB - The purpose of our study was to analyze the immunohistochemical expression of two MMR system proteins at different steps of neoplastic progression within the squamous cervical epithelium. We compared cases showing normal histologic appearance with those affected by low and high-grade squamous intraepithelial lesions and invasive cervical carcinoma. We investigated formalin-fixed and paraffin-embedded tissue specimens obtained from 83 selected patients (55 with preinvasive neoplastic lesions and 28 with invasive squamous cervical carcinoma) for the expression of hMSH2 and hMLH1 at the immunohistochemical level. We also included 30 patients with histologically normal cervix as a control group. Epithelial cells of CIN lesions showed a significant increase in the expression of both hMLH1 and hMSH2 proteins compared to non-neoplastic squamous epithelium (p < 0.0001). The cases of invasive carcinoma showed a positivity for hMLH1 protein that was statistically lower than that for non-neoplastic cells (p = 0.0009) and that for cases with CIN (p < 0.0001). Positivity for hMSH2 protein was higher than that for normal epithelium (p = 0.0007), but lower than that for preinvasive lesions (p = 0.0001). Preinvasive lesions showed increased expression of both proteins if compared with normal esocervical epithelium. Neoplastic stromal invasiveness is associated with a significant loss of hMLH1 function. PMID- 15807308 TI - A new in vitro model for analyzing the biological behavior of well-differentiated squamous cell carcinoma. AB - A suitable model analyzing the behavior of well-differentiated squamous cell carcinoma has not yet been established. We tried to establish such a system using a reconstructed oral mucosa, in which T3M-1 squamous cell carcinoma cells were cultured on 3T3 fibroblast-containing collagen gel. Fibroblasts promoted the stratification and keratinization of T3M-1 cells. During growth, the Ki-67 index of T3M-1 cells with fibroblasts was higher than that of T3M-1 cells alone. Fibroblasts increased the expression of involucrin, a differentiating marker of keratinocytes, in T3M-1 cells. They also promoted the invasion of T3M-1 cells into the gel. When T3M-1 cells alone were cultured in a fibroblast-conditioned (FC) medium, the fibroblast-induced phenomena mentioned above were almost replicated. In addition, epidermal growth factqr (EGF) promoted T3M-1 cells growth, but not the invasion. cDNA microarray analysis showed that FC medium increased the expression of EGF receptor and several other mRNAs of T3M-1 cells. The data suggest that T3M-1 cells, under cancer-stromal fibroblast interaction, undergo invasive growth with their well-differentiated squamous phenotype, and that this interaction may be mediated partly by soluble molecules (e.g., EGF) in an autocrine or paracrine pathway. Our system will probably provide a useful model for analyzing the biological behavior of well-differentiated squamous cell carcinoma. PMID- 15807309 TI - Improved detection of mycobacterial DNA by PCR in formalin-fixed, paraffin embedded tissues using thin sections. AB - PCR is a unique methodology allowing for the sensitive detection of mycobacterial DNA-sequences in cases in which no fresh material can be obtained for classic analyses. Despite the limitations of this technique, for example the less satisfactory quality of DNA from paraffin-embedded specimens and the high effort necessary to control contamination, PCR still represents a useful additional tool for routine diagnostic examinations of mycobacterial infections. Fragmentation of the DNA extracted from formalin-fixed, paraffin-embedded samples on the one hand and the rigid cell wall of mycobacteria on the other hand are obstacles to detecting the DNA of these microorganisms by PCR. Here, we describe a simple mechanical procedure that allows us to improve the detection of mycobacterial DNA with the use of thin (1 microm) sections instead of thicker sections. This could be explained by a gentle, mechanical opening of the acid fast mycobacterial cell wall. Thus, even the application of heat/cold shock treatments is not necessary. This inexpensive fast procedure can also be used for the detection of other infectious agents. PMID- 15807310 TI - Fluorescent in situ hybridization in liver cell touch preparations from autopsy. AB - The objective of the present study was to develop a simplified and low-cost protocol for the investigation of congenital anomalies of chromosomal etiology by fluorescent in situ hybridization (FISH) using probes for chromosomes X, 18, 13/21 in liver cell touch preparations obtained from autopsies performed at the University Hospital of the Faculty of Medicine of Ribeirao Preto. Liver touch preparations were obtained from 11 autopsy cases and fixed in 95% ethanol or methanol:acetic acid (3:1). The FISH technique was carried out according to the protocol of Pinkel with modifications, using probes for chromosomes X, 18, 13/21. There was no significant difference in labeling intensity, quantity of nuclei, or number of signals present per nucleus between the materials fixed with the two fixatives. Similar results were obtained with different times of storage up to 14 months at -20 degrees C. We concluded that the use of touch preparations pretreated with acetic acid and fixed in 95% ethanol represents an efficient, practical, and low-cost method of cell preparation for FISH analysis. PMID- 15807311 TI - A case of primary ductal adenocarcinoma of the lacrimal gland: histopathological and immunohistochemical study. AB - We encountered primary ductal adenocarcinoma of the lacrimal gland in a 67-year old Japanese man. To the best of our knowledge, only three cases of primary ductal adenocarcinoma of the lacrimal gland have been reported in the literature. The patient was admitted because of visual disturbance, and a mass measuring about 3 cm in diameter was revealed in the right orbit. The mass was resected, and primary ductal adenocarcinoma of the lacrimal gland was diagnosed histopathologically. He died from recurrence at the primary site and metastasis to the brain, lungs, liver, common bile duct, and pancreas 2 years and 10 months after surgery although adjunctive orbital radiotherapy was given. Immunohistochemically, the characteristics of cancer cells were similar to those of salivary duct carcinoma, namely positivity for cytokeratin (CK) 7, 10, 17, 18, 19, and 34betaE12, and negativity for CK20. It was not clear whether the ductal adenocarcinoma originated from the ductal or acinar epithelium of the lacrimal gland, because the immunohistochemical features of both epithelia were identical. PMID- 15807312 TI - Hepatocellular carcinoma occurring in alagille syndrome. AB - Hepatocellular carcinoma only rarely occurs in Alagille syndrome. Here, we report on three cases of hepatocellular carcinoma associated with Alagille syndrome. All three patients were boys and presented with jaundice. In addition, they had the characteristic facial appearance of Alagille syndrome with cardiac, vertebral, and eye anomalies, and all had passed acholic stools from the neonatal period. Liver biopsies were diagnosed as bile duct paucity, compatible with Alagille syndrome in two cases, but the third case showed marked bile duct proliferation at the initial liver biopsy when 7 months old, which made diagnosis difficult. Eventually, all three cases progressed to biliary cirrhosis and hepatocellular carcinoma, which occurred at 17 months, 4 years, and 7 years, respectively. Because of the unusual liver histology and early onset of hepatocellular carcinoma, careful clinicopathologic correlations and close monitoring are required for the diagnosis of Alagille syndrome and for the early detection of hepatocellular carcinoma. PMID- 15807314 TI - Pure (non-papillary) serous cystadenoma of the epididymis: a histologic and immunohistochemical study. AB - A 44-year-old man presented with painless right scrotal swelling of 2 years duration. A cystic tumor strictly attached to the head of the epididymis was surgically resected. The pathologic examination revealed a unilocular cyst with a thin fibrous capsule, lined by ciliated cubical or cylindrical columnar cells, mostly arranged in a single layer. No papillary projection could be detected. Immunohistochemical staining was positive for epithelial membrane antigen, low- and high molecular weight cytokeratins, progesterone receptor, vimentin, and S 100 protein, but was negative for carcinoembryonic antigen, CD10, p53 protein, and calretinin. Single MIB-1 positive cells were noted. Histologic and immunohistochemical features suggest a Mullerian origin or differentiation. The lesion was diagnosed as pure serous cystadenoma of the epididymis, possibly originating from vestigial remnants of the Muller duct in male. The differential diagnosis to spermatocele and adenomatoid tumor of the epididymis is discussed. PMID- 15807313 TI - A persistently recurring peri-urethral soft tissue lesion of the vulva. AB - Various mesenchymal lesions, some of which have only recently been characterized, may affect the vulva. Because of their apparently shared origin from vulvovaginal mesenchyme, these lesions can resemble one another superficially and can be challenging diagnostically. Clinically, benign lesions and tumor-like conditions may mimic a malignant process because of hypercellularity, mitotic activity, and rapid growth. We report on a 84-year-old diabetic and hypertensive woman who had previously undergone skinning vulvectomy for lichen planus and an embolectomy of the left femoral artery. The patient presented with a rapidly growing vulvar mass. The biopsy showed spindle cell proliferation in a loose myxoid stroma with granulation tissue and a mixed inflammatory infiltrate. Based on histology and immunohistochemistry, the initial diagnosis was that of a benign lesion. In view of the persistent recurrence of the lesion and the absence of metastatic spread, we conclude that the lesion has a low-grade malignant potential and suggest the diagnosis of florid cellular reactive pseudosarcomatous myofibroblastic proliferation. PMID- 15807315 TI - Back to the future: rebuilding public health infrastructure. PMID- 15807316 TI - Public health and law enforcement: intersecting interests, collegiality and cooperation. PMID- 15807317 TI - Redefining public health. PMID- 15807318 TI - Smart growth for community development. PMID- 15807319 TI - Commercial speech and the public's health: regulating advertisements of tobacco, alcohol, high fat foods and other potentially hazardous products. PMID- 15807320 TI - Legal frameworks for chronic disease prevention. PMID- 15807321 TI - Reducing underage drinking: the role of law. PMID- 15807322 TI - The pros and cons of litigation in public health. PMID- 15807323 TI - New partners in community legal preparedness: public health and the health care bar. PMID- 15807324 TI - Public health in court: who's to judge? PMID- 15807325 TI - Jails, prisons, and your community's health. PMID- 15807326 TI - Public health and law enforcement: future directions. PMID- 15807327 TI - School-based policies: safety and injury liability. PMID- 15807328 TI - Innovative legal tools to prevent obesity. PMID- 15807329 TI - Elder abuse and the law: new science, new tools. PMID- 15807330 TI - Legal approaches to regulating internet tobacco sales. PMID- 15807331 TI - How drug courts reduce substance abuse recidivism. PMID- 15807332 TI - Living with the HIPAA privacy rule. PMID- 15807333 TI - Are you ready for the next outbreak? An exercise in legal preparedness. PMID- 15807335 TI - Quarantine: voluntary or not? PMID- 15807334 TI - Private property in public health emergencies. PMID- 15807336 TI - Globalization, public health, and international law. PMID- 15807337 TI - Lawyers as advocates in public health practice. PMID- 15807338 TI - Interdisciplinary contributions to public health law. PMID- 15807339 TI - How states are using the Turning Point Model State Public Health Act. PMID- 15807340 TI - Connecting public health law with science. PMID- 15807341 TI - Methamphetamine: tools and partnerships to fight the threat. PMID- 15807342 TI - Bernard Dickens: bespoke public health, law and ethics. PMID- 15807343 TI - How litigation can promote product safety. PMID- 15807344 TI - Using litigation to make public health policy: theoretical and empirical challenges in assessing product liability, tobacco, and gun litigation. PMID- 15807345 TI - Pandemic Influenza: public health preparedness for the next global health emergency. PMID- 15807346 TI - Towards progress in resolving dilemmas in international research ethics. PMID- 15807347 TI - Research involving humans: a time for change? PMID- 15807348 TI - The evolution of research ethics: the current international configuration. PMID- 15807349 TI - Regulating Research and experimentation: a view from the U.K. PMID- 15807350 TI - Institutional conflicts of interest: protecting human subjects, scientific integrity, and institutional accountability. PMID- 15807351 TI - Legal and ethical approaches to stem cell and cloning research: a comparative analysis of policies in Latin America, Asia, and Africa. PMID- 15807352 TI - Leopards in the temple: restoring scientific integrity to the commercialized research scene. PMID- 15807353 TI - American bioethics and human rights: the end of all our exploring. PMID- 15807354 TI - What are the limits of bioethics in a culturally pluralistic society? PMID- 15807355 TI - Challenging themes in American health information privacy and the public's health: historical and modern assessments. PMID- 15807356 TI - The changing legal and conceptual shape of health care privacy. PMID- 15807357 TI - The ELSI genetics regulatory resource kit: a tool for policymakers in developing countries. PMID- 15807358 TI - Disputes about the withdrawal of treatment: the role of the courts. PMID- 15807359 TI - An appraisal of abortion laws in southern Africa from a reproductive health rights perspective. PMID- 15807360 TI - Law and clinical Research--from rights to regulation? An English perspective. PMID- 15807361 TI - Commentary: Social-ethical values issues in the political public square: principles vs. packages. PMID- 15807362 TI - Storing newborn blood spots: modern controversies. PMID- 15807363 TI - What conditions justify risky nontherapeutic or "no benefit" pediatric studies: a sliding scale analysis. PMID- 15807364 TI - Shared decision-making and the lower literate patient. PMID- 15807365 TI - Ethics in the practice of health law. PMID- 15807366 TI - Supreme Court rules HMO suits are preempted. PMID- 15807367 TI - The demise of Bayh-Dole protections against the Pharmaceutical Industry's abuses of government-funded inventions. PMID- 15807368 TI - Symmetry cues for matching mirrored objects. AB - The study explores cues for matching pairs of objects. These objects were arranged into mirror-symmetrical displays and the task was to judge whether a pre specified 180 degree rotation around the X, Y or Z axis carries one object into the mirror object. For some rotations, the object's mirror symmetry (M) and, for other rotations, the object's point-symmetry (P) could serve as a cue. The matching results suggest that M is a better cue than P, say M > P. Various attempts are made to explain this effect. The most promising one focuses on simplest structural object representations as these capture M and not P. It is furthermore plausible that M captured by reference frames at high hierarchical representation levels, say M1, serves as a better cue than M captured at low levels, say M2. The prediction M1 > M2 > P merely applies to the open surface objects in the experiment. For the closed solid objects in the experiment the expectation is M1 = M2 = P. Both predictions roughly agree with the accuracy and reaction-time data. The results suggest the perceptual relevance of representation cues and, aditionally, that cues stemming from reference frames at higher hierarchical representation levels are more effective than those from lower levels. PMID- 15807369 TI - Adelson's tile and snake illusions: a Helmholtzian type of simultaneous lightness contrast. AB - Adelson's tile, snake, and some other lightness illusions of the same type were measured with the Munsell neutral scale for twenty observers. It was shown that theories based on low-level luminance contrast processing could hardly explain these illusions. Neither can those based on luminance X-junctions. On the other hand, Helmholtz's idea, that simultaneous lightness contrast originates from an error in judgement of apparent illumination, has been elaborated so as to account for the tile and snake illusions as well as other demonstrations presented in this report. PMID- 15807370 TI - A random-ray model for speed and accuracy in perceptual experiments. AB - We present a 'random ray' model to describe Yes/No reaction times (RTs) and errors in perceptual experiments. The ray model is analogous to a random walk, but it is computationally simpler, requiring only elementary geometry. Ray parameters control the drift rates to the Yes and No decision boundaries, bias, and a termination or 'time-out' rule. Rays are normally distributed, but predicted RT distributions are skewed by projection onto the boundaries. Model parameters can be estimated directly from the 16th, 50th, and 84th percentiles of the RT distributions on hit, correct rejection, false alarm, and miss trials, if the data satisfy three easily testable constraints. Examples are given from visual search and object recognition. PMID- 15807371 TI - Contrast sensitivity functions to stimuli defined in Cartesian, polar and hyperbolic coordinates. AB - Recent electrophysiological studies indicate that cells in the LGN, V1, V2, and V4 areas in monkeys are specifically sensitive to Cartesian, polar and hyperbolic stimuli. We have characterized the contrast sensitivity functions (CSF) to stimuli defined in these coordinates with the two-alternatives forced-choice paradigm. CSFs to Cartesian, concentric, and hyperbolic stimuli have had similar shapes, with peak sensitivity at approximately 3 c/deg. However, the Cartesian CSF peak sensitivity has been at least 0.1 log units higher than that to stimuli in any other coordinate system. The concentric-Bessel CSF has a low-pass shape, peaking at 1.5 c/deg or below. The radial CSF has a bell shape with maximum sensitivity at 8 c/360 degrees. Only the concentric-Bessel CSF could be explained in terms of the components of maximum amplitude of the Fourier transform. Neural models, which in previous studies predicted the responses to Cartesian and polar Glass patterns, failed to account for the full CSFs data. PMID- 15807372 TI - Thresholds vary between spatial and temporal forced-choice paradigms: the case of lateral interactions in peripheral vision. AB - Psychophysicists use spatial or temporal two-alternative forced-choice (2AFC) paradigms interchangeably. Thus, experiments with the same general goal are carried out using one or the other paradigm by distinct or even the same research groups. For example, this situation has occurred both in studies on visual sensitivity in dyslexia and in studies on lateral interactions in peripheral vision. Conflicting results in either field (e.g. whether or not dyslexics have a visual deficit and whether or not peripheral detection is facilitated by the presence of flankers) appear to be resolved on the surmise that spatial and temporal 2AFC paradigms indeed produce different results. We designed experiments in which peripheral detection thresholds for Gabor patches (in the presence or absence of suprathreshold flankers) could be measured using completely equivalent spatial and temporal 2AFC paradigms so that any resultant difference can be unequivocally attributed to the effect of the paradigms themselves. The results showed that spatial 2AFC renders significantly lower sensitivity than temporal 2AFC when the target is presented along with suprathreshold flankers, but about the same sensitivity as temporal 2AFC when the target is presented alone. In the end, this resulted in statistically significant facilitation in peripheral vision only when measured with temporal 2AFC. Separate experiments at each of several peripheral locations revealed that the presence and magnitude of this effect varies not only with psychophysical paradigm but also with retinal locus. PMID- 15807373 TI - Mechanical properties of the take-off leg as a support mechanism in the long jump. AB - The aim of this study was to establish the functions of the support leg in the long jump take-off with a three-element mechanical model spring, damper, and actuator The take-off motions of eleven male long jumpers, with personal bests from 6.45 to 7.99 m, were videotaped at 250 Hz and ground reaction forces were simultaneously recorded at 1 kHz. A two-dimensional 14-segment linked model was used to collect basic kinematic parameters. The spring, damper and actuator forces were determined from the displacement and velocity of the centre of mass and from ground reaction forces. Large spring and damper forces were exerted, and absorbed the impact force immediately after the touch-down. The spring force was also exerted from 25 to 75% of the take-off phase. The actuator force was dominant in the latter two-thirds of the take-off phase. Statistically significant correlations were found between the spring force impulse and the knee flexion during the take-off phase (r = 0.699, p < 0.05), and between the knee flexion and the angular velocity of the thigh at the touch-down (r = 0.726, p < 0.05). These results indicated that the jumper should retain less flexion of the take-off leg knee to increase the spring force, after a fast extension of the hip, and use a more extended knee at the touch-down to prevent excessive knee flexion. PMID- 15807374 TI - Off-side front foot drives in men's high performance cricket. AB - We investigated the techniques used by nine right-handed, international batsmen to perform front foot off-side drives in first class and international matches. All strokes were captured using two synchronised high-speed video cameras; nine were selected for kinematic analysis. These movement sequences were then manually digitised at a sampling frequency of 125 Hz using APAS motion analysis software. The results of this study indicated that the batsmen used movement patterns that enabled important aspects of stroke production, such as the front stride and the downswing of the bat, to be delayed so that additional information from ball flight could be assimilated. Front upper limb segments were constrained to work in a unitary fashion, with the peak horizontal end point speed of each segment occurring almost simultaneously just before impact. It has been suggested that these strategies serve to enhance stroke accuracy. Other aspects of their techniques included a distinctively looped bat path, a front foot placement that occurred only just before impact, and a front ankle that was positioned well inside the line of the ball at impact. Various technical parameters, such as the alignment of the trunk relative to ground and the continuous flow of the bat between the backswing and downswing, were similar to findings in previous batting research. Other characteristics of stroke production not previously addressed, including the path of the bat and the timing of the front stride, may challenge some long held beliefs evident in current coaching literature. PMID- 15807375 TI - A comparison of the ball rebound characteristics of wooden and composite cricket bats at three approach speeds. AB - The primary aim of this study was to compare the rebound characteristics of wooden and composite cricket bats. The rebound characteristics of two 'experimental' bats manufactured from composite material were compared with three English willow bats and one Kashmir willow bat. The bats were tested using a specially designed testing rig, which propelled a 156 g Kookaburra cricket ball at three impact speeds: fast-medium, 67 km x h(-1); fast, 101 km x h(-1); and express, 131 km x h(-1) on to the bats mounted in position so that the ball impacts occurred at the position where the blade of the bats was the thickest. The rebound characteristics of the bats were calculated by measuring the approach and rebound speeds of the ball as it passed through a light beam positioned a short distance away from the point of impact. The statistical software package SAS was used to test for significant differences (p < 0.05) between the average rebound characteristics of the bats. Further, Scheffe's method was used as a post hoc comparison to determine whether differences existed between the composite and willow bats. When the composite and traditional willow bats were compared, the results showed no significant differences between the three average approach speeds, while the composite bats showed significantly smaller rebound speeds and coefficient of restitution at all three approach speeds. Thus, the rebound characteristics of the composite bats were significantly less than the traditionally designed English willow wooden bats and would not enhance performance by allowing the batsman to hit the ball harder, assuming all other factors, such as bat speed, mass distribution and the impact point, were the same for the bats. Further study is required to determine the physical properties of composite and wooden bats to enhance their impact characteristics. PMID- 15807376 TI - Equipment set-up among Olympic sprint and slalom kayak paddlers. AB - The anthropometry and equipment set-up for sprint (31 male; 11 female) and slalom (12 male; 12 female) kayak paddlers who competed at the 2000 Olympic Games in Sydney were measured in the 15 day period before competition. This paper provides normative data for equipment set-up in these sports, as well as information about differences in rigging and paddle dimensions between sprint and slalom kayak paddlers. These differences were consistent for both male and female athletes, with sprint paddlers seated higher and using longer paddles with longer, though narrower, blades (p < 0.0001). Among male sprint paddlers, only minor differences in equipment set-up were found between competitors ranked in the top 10 places compared to the rest of the field. Considering all male paddlers initially, then sprint paddlers alone, significant (p < 0.01) regression equations were developed for the prediction of foot bar distance (r2 = 0.482 and 0.589 respectively) and hand grip distance (r2 = 0.400 and 0.541 respectively). The process of fine tuning equipment set-up often requires hours of practice with subjective feedback from the athlete. The normative data presented in this paper should assist coaches with this process as their athletes evolve toward their individual optimum set-up. PMID- 15807377 TI - Full-body kinematic characteristics of the maximal instep soccer kick by male soccer players and parameters related to kick quality. AB - Many studies try to understand fundamental soccer skills, most focusing on kicking. However, a full picture of an efficient kick remains incomplete owing to constraints of test designs and difficulties that arise in synchronizing and analyzing information generated by multiple assessment techniques. Previous scientific studies may be generally categorized as: two-dimensional kinematic analysis using high-speed cameras, muscle activity studies using electromyography, three-dimensional analysis of the kicking-leg using a partial body model, and kinetic studies using force measurements and modelling. No existing inquiries have used full-body three-dimensional motion capture and modelling to examine kicking. The current study remedies this deficiency and defines a full-body model capable of revealing more detailed characteristics of kicking. Additionally, it reveals effects of long-term training by comparing novices with skilled athletes and explores new parameters that have potential to aid quantitative evaluations of skill. Results show effective upper-body movement to be a key factor in creating better initial conditions for a more explosive muscle contraction during kicking. It permits a more powerful quasi whip-like movement of the kicking leg. Finally, the timely change of distance between the kick-side hip and the non-kick-side shoulder provides a quantitative means of measuring kick quality. PMID- 15807378 TI - Coaching points for the technique of the eggbeater kick in synchronized swimming based on three-dimensional motion analysis. AB - We investigated coaching points for the technique of the eggbeater kick in synchronized swimming. The movements of the eggbeater kick for six female synchronized swimmers, all the top-ranking members at the Olympics and World Championships, were analyzed by using three-dimensional motion analysis. By comparing the movements of higher eggbeater-skilled swimmers with lower eggbeater skilled swimmers, the coaching points for techniques of the eggbeater kick in synchronized swimming are clarified as follows. First, to hold the knees as high and as near the water surface as possible and to keep the heels close to the hips. Secondly, to keep the knees as wide as possible. Thirdly, to kick sideways producing lift force, and not to kick downward. Fourthly, to pull up the heels to nearly the water surface at the out-kick, with strong internal rotation of the thighs. Lastly, to move the feet as if treading water, with strong dorsiflexion and plantar flexion and adduction of the foot; that is to move the feet like the motion of the hands when sculling. Moreover, the movements of higher eggbeater skilled swimmers can be termed 'horizontal kick type' and the movements of lower eggbeater-skilled swimmers can be termed 'vertical kick type'. PMID- 15807379 TI - Variations of stroking parameters associated with 200 m competitive performance improvement in top-standard front crawl swimmers. AB - The aim of this study was to analyse the variations of stroking parameters (speed, stroke length, stroke rate, and stroke index) associated with the 200 m front crawl competitive performance improvement. Two races completed by 17 top swimmers were analysed in the 200 m freestyle final of French or European championships, each final being separated by two years. All the swimmers' performances were bettered in the second race (mean +/- SD: 113.44 +/- 2.50 vs 111.78 +/- 2.71 s; p < 0.01) and were associated with a significant increase of stroke rate without variation of average stroke length and stroke index values (p > 0.05). Swimmers emphasized the first part of the race, with higher speed in the first three lengths, higher stroke rate in the first two lengths and lower stroke length in the first one. Stroke length and stroke rate variations were highly correlated (r = 0.98; p < 0.05). In 11 of the 17 swimmers, the improvement was concomitant with a decrease in stroke length and an increase in stroke rate. Only one swimmer's improvement was associated with a substantial increase in stroke length. These results highlighted that stroke length and stroke index cannot be considered as the only parameters linked to improvement in a 200 m crawl in adult swimmers competing at high standard. Moreover, an increase in stroke rate associated with a slight decrease in stroke length should not be considered as ineffective, especially at top standard. PMID- 15807380 TI - Criterion validity of an isokinetic dynamometer to assess shoulder function in tennis players. AB - Isokinetic dynamometry is used in the assessment and rehabilitation of shoulder function in tennis players. The aim of this study was to validate a newly installed Biodex III Isokinetic Dynamometer for internal and external shoulder rotation. A non-injured male performed this movement which replicates an integral section of the tennis service action (Cohen et al., 1994). Three maximal trials were performed at six randomly assigned angular velocities (0.52, 1.05, 1.57, 2.09, 2.62 and 3.14 rad x s(-1)) with 90 s rest between each set. Trials were recorded using an on-line motion analysis system and kinematic data were obtained. Kinematic angular displacement of the lever arm was slightly greater (> or = 0.01 rad x s(-1)) than for the isokinetic dynamometer at all test angular velocities. Mean angular velocities from the kinematic data were almost identical to those from the isokinetic dynamometer but less than the target values, and this difference (e.g. 2.55 rad x s(-1) at 3.14 rad x s(-1)) was greater at higher angular velocities owing to the greater acceleration phases required. Peak angular velocity was similar on the isokinetic dynamometer to the target values, but substantially less than those from the kinematic data (e.g. 0.68 rad x s(-1) at 0.52 rad x s(-1)). This suggests that the isokinetic data are over-smoothed and may mask important information. In summary, criterion validity of this isokinetic dynamometer is supported for displacement and mean angular velocity, but not for peak angular velocity. Shoulder rotations in tennis are often at angular velocities greater than the Biodex can reproduce, but for the purpose of monitoring the shoulder strength and range of motion of tennis players and providing safe functional rehabilitation, the use of slower angular velocities is acceptable. PMID- 15807381 TI - [Disribution of Streptococcus pneumoniae serotypes and serogroups among patients with invasive pneumococcal diseases in the Czech Republic in 1996-2003: background data for vaccination strategy]. AB - STUDY OBJECTIVE: To map distribution of Streptococcus pneumoniae serotypes and serogroups in the Czech Republic in 1996-2003 in order to provide background data for effective strategy of vaccination with type specific pneumococcal conjugated vaccine. METHODS: Quellung typing with sera from the Statens Serum Institute (Denmark) was used to differentiate 1553 S. pneumoniae strains isolated from patients with invasive pneumococcal infections in the Czech Republic. RESULTS: Strains of serotypes 3, 19F, 9V, 23F, 1, 14 and 4 were most frequently isolated from clinical specimens (blood, cerebrospinal fluid, autopsy specimens, lung puncture aspirate, bronchoalveolar lavage fluid, sputum) of patients with invasive pneumococcal infections. Some serotypes were detected regardless of patient age (1, 3, 6A, 9V, 14, 19F and 23F), while serotype 6B was most frequent in the age group 0-2 years, serotypes 4, 6B, 7F and 18C were typically found in the age group >2-5 years and types 4, 7F, 8, 9N, 10A and 11A were common in patients aged >5 years. The 7-valent conjugated pneumococcal vaccine (PNCMR7) covers 63.1%, 62.3% and 34.3% of the serotypes identified in 152 patients aged 0 2 years, 168 patients aged >2-5 years and 1233 patients aged >5 years, respectively. It includes 38.3% of the serotypes identified among 1055 strains isolated from blood, cerebrospinal fluid and autopsy specimens regardless of patient age. Over the period moni- tored, the percentage of strains allocated to one of the serotypes included in the vaccine increased from 45.5% to 70.6%, from 54.3% to 38.7% and from 21.8% to 38.7% in patients aged 0-2 years, >2-5 years and >5 years, respectively, and from 30.0% to 39.4% for the isolates from blood, cerebrospinal fluid and autopsy specimens. A similar trend was found in the distribution of serogroups. These shifts can also be ascribed to the continuous increase in the percentages of strains of serotypes 4 (from 1.0% to 8.9%) and 9N (from 0.0% to 5.9%). CONCLUSION: The PNCRM7 vaccine is recommended for use in the Czech Republic in spite of the lower prevalence of vaccination serotypes among strains isolated from Czech patients with invasive pneumococcal infections which is a limitation to its efficacy. Nevertheless, results of the longterm monitoring of serotype distribution in the Czech Republic are suggestive of the need for inclusion of types 1 and 3 in the vaccine in the future. PMID- 15807382 TI - [Inhibition of hemolytic activity of Staphylococcus aureus 3-hemolysin by an exosubstance produced by some Enterococcus faecalis strains]. AB - OBJECTIVE: Accidentally, some Enterococcus faecalis strains were found to inhibit the hemolysis caused by Staphylococcus aureus beta-hemolysin. The study objectives were to define the conditions under which this phenomenon appears and to characterize the inhibitory agent. MATERIALS AND METHODS: Ninety-one wild strains and five type strains of E. faecalis and nine wild E. faecium strains were tested for antihemolytic activity on blood agar with 5% sheep erythrocytes prepared using the following agar bases: Columbia Agar Base (BBL, 211124), Columbia Agar (Bio Merieux, 51026), Blood Agar Base (Bio-Rad, 64524), Blood Agar Base (Columbia) (Ferosa, 1-034), Columbia Blood Agar Base (HiMedia, M114), Blood Agar Base No. 4 (Imuna), Columbia Agar Base (Merck, 10455) and Columbia Agar Base (Oxoid, CM331). The exosubstance produced by E. faecalis strains was isolated by means of acetone precipitation and was exposed to elevated temperature and trypsin. RESULTS: Thirty-three (36%) out of 91 wild strains of E. faecalis produced the substance inhibiting hemolysis caused by Staphylococcus aureus beta hemolysin. Two out of five E. faecalis type strains and none of E. faecium strains appeared to be producers of the same substance. Inhibition of the hemolysis was observed on all of the media tested, being the most pronounced with the use of Blood Agar Base No. 4 (Imuna) and Columbia Agar Base (BBL) at pH 7.0 7.5 when cultured at 37 degrees C for 24 hours. The exosubstance lost its inhibitory effect after exposure to 70 degrees C for 15 min and to 0.1% trypsin for 30 min. The inhibitory potential against staphylococcal hemolysis correlated with the ability of strains to produce protease. CONCLUSION: About one third of wild E. faecalis strains produce a protease-like substance that seems proteinaceous in nature and has an inhibitory effect on the hemolysis caused by Staphylococcus aureus beta-hemolysin. PMID- 15807383 TI - [Poliomyelitis surveillance in the Czech Republic from the start of vaccination to the certification of eradication in the European Region]. AB - Poliomyelitis anterior acuta is an acute infectious disease caused by polioviruses of three antigenic types. First epidemics of poliomyelitis emerged at the end of the 19th century. The World Health Organization launched the poliomyelitis eradication program in 1988. The incidence of poliomyelitis in the world decreased from 350,000 cases in 1988 to 1918 cases in 2002 when poliomyelitis eradication was certified in three WHO regions, the European Region (2002), American Region (1994) and West Pacific Region (2000). Systematic clinico virological surveillance of poliomyelitis has been carried in the Czech Republic since 1961, including annual vaccination campaigns with living OPV vaccine, clinical screening, virological screening of clinical specimens and sewage water (environmental) samples and sera screening within serological surveys mapping the vaccination immunological efficacy. From 1961 to 2003, 21,423 stool specimens of vaccinated healthy children, 62,440 stool specimens of patients, 6250 cerebrospinal fluid specimens and 2100 throat swab specimens were screened. Within the outdoor environment surveillance, 15,460 sewage water samples were analysed. From 1995 to 2003 129 cases of acute flaccid paresis were investigated in children under 15 years of age and 28 stool samples from their contacts were screened. Over the same period, 1280 sewage water samples from refugee camps were analysed. For serological surveys, about 60,000 sera from healthy individuals of all age categories were investigated. No case of paralytic poliomyelitis has been reported and no wild virus has been isolated in the Czech Republic since 1961. PMID- 15807384 TI - [Longitudinal epidemiology of leptospirosis in the Czech Republic (1963-2003)]. AB - After higher rates of occupational or leisure diseases recorded in the Czech Republic in the second half of the last century it was found that the last climatic changes and catastrophic floods of 1997 and 2002 were followed by outbreaks of leptospirosis as the only re-emerging postflood infection. While in tropics and subtropics the monsoon season is typically followed by highly increased rates of leptospirosis in humans, even, with fatal outcomes, this phenomenon remained unknown under the climatic conditions of the Czech Republic where human leptospirosis has been reported rather sporadically, mostly in natural foci of infection, and its incidence rate is usually about 0.3% per 100,000 population. Nevertheless, after the unexpected vast floods of 1997 and 2002 that also afflicted natural foci of leptospirosis, the rates of reported and serologically confirmed cases of leptospirosis in the Czech Republic were three times as high as usual with the specific morbidity reaching 0.9 case per 100,000 population. In 1997, as many as 7156 persons were tested for leptospirosis in the Czech Republic: the disease was diagnosed and reported in 94 patients and in 2002, 92 out of 4999 persons tested were diagnosed with leptospirosis. Two thirds of these cases were from inundation areas, half of them being directly associated with floods (exposure to residual water, flood mud in cellars, etc.). Four case of Weil disease reported in 1997 were fatal. As many as 41 deaths from Weil disease have been reported in the Czech Republic since 1963 when the disease became reportable. PMID- 15807385 TI - [HCV genotyping by combination of the Cobas Amplicor HCV 2.0 test and the reverse hybridization Versant HCV Genotype Assay]. AB - Genotyping of hepatitis C virus (HCV) is of relevance to scheduling the treatment of patients with chronic hepatitis C (VHC), making their prognosis and monitoring the treatment efficacy. A set of 62 sera testing HCV RNA positive in Cobas Amplicor HCV 2.0 test (CA) were genotyped using Versant HCV Genotype Assay (LiPA) Bayer, i.e. the reverse hybridization method, with the CA amplified product being directly used in the assay. Fifty-six out of 57 samples reactive in reverse hybridization (92%) were genotyped. One sample showed a profile differing from any genotype, five samples were not reactive and one sample was not tested within this study design. Two out of five non-reactive sera and one non-tested serum could be genotyped by nested PCR based reverse hybridization. It can be concluded that the CA product resulting from one-step HCV RNA amplification is suitable for use in genotyping by reverse hybridization. The CA product based genotyping procedure is easier to perform, less time-consuming and less costly. The nested PCR based procedure could be used for typing of sera with lower HCV concentrations nontypeable with the combination of CA and Versant HCV Genotype Assay. Forty-eight selected samples were typed not only by reverse hybridization but also by a serological kit Murex HCV Serotyping 1-6 Assay (Abbot Murex). Thirty-seven (77%) of these sera, including all of three sera negative in reverse hybridization, appeared typeable by this kit. Although less sensitive, serotyping may be of relevance to typing of sera with low HCV levels or not containing detectable viral NA which are nontypeable by reverse hybridization. Thirty-three sera appeared genotypeable by both of the methods tested with the results being in good agreement. In two cases only the serotyping method revealed one more type of virus (mixed genotype) compared to the reverse hybridization. PMID- 15807387 TI - The phenomenon of preconditioning today. PMID- 15807388 TI - Early statin therapy in patients with acute coronary syndrome. PMID- 15807389 TI - Vascular wall shear stress: basic principles and methods. PMID- 15807390 TI - Strategies for the invasive treatment of multivessel coronary artery disease. PMID- 15807391 TI - Histological study of arterial and venous grafts before their use in aortocoronary bypass surgery. AB - INTRODUCTION: In this study we investigated the morphology of grafts from the internal thoracic artery and the great saphenous vein, before their use in aortocoronary bypass surgery, in order to draw conclusions concerning their suitability and viability. MATERIAL AND METHODS: Sections of grafts from the great saphenous vein and left internal thoracic artery obtained for use in bypass surgery were examined using light microscopy and transmission and scanning electron microscopy. RESULTS: The histological changes in the walls of the vessels were classified as acute or chronic. The acute lesions concerned the endothelium and the subendothelial layer. There was extensive necrosis of endothelial cells, resulting in the basement membrane being left uncovered and becoming the target of blood cells. The endothelial necrosis was accompanied by subendothelial oedema and focal destruction of the inner elastic lamina of the internal thoracic artery. The chronic lesions affected mostly the venous grafts and included the presence of distinct atheromatous plaques or thickening of the intima and media. CONCLUSIONS: The combination of ischaemic and chronic atheromatous lesions in bypass grafts may contribute to a decrease in their viability, especially in the case of venous grafts. PMID- 15807392 TI - Determinants of heart rate turbulence after ventricular premature beats in healthy volunteers. AB - INTRODUCTION: A decline in the acceleration of the heart rate (HR, heart rate turbulence, HRT) after a ventricular premature capture is associated with increased mortality in patients suffering from coronary artery disease. The physiological properties of HRT have not been evaluated in a large human cohort. METHODS: In 95 healthy individuals, HRT parameters onset (TO) and slope (TS) as well as the turbulence timing (TT) were calculated from 24-hour Holter ECGs. With the help of a simple, linear, weighted regression model, gender specific differences of TO and TS were compared. A multiple linear regression model served to evaluate the influence of age and the basic HR preceding the ventricular premature contraction (VPC) on HRT. RESULTS: The median of TT is present in regression line #5. We discovered that, in men and women, TO is reduced as basic HR rises (p<0.01). In contrast, analysis of TS showed a divergence: in men, TS declines as basic HR increases. However, basic HR modifies TS in women to some extent (p<0.01). A multiple, linear regression model revealed a decrease of HRT with increasing age in men. CONCLUSIONS: The acceleration of HR after a ventricular premature beat occurred within the first 11 beats in more than 75% of our healthy individuals. An increased HR prior to VPC affects HRT in men. Increasing age leads to an attenuation of HRT in men and to a reduction of TO in women. These results emphasise the significance of the physiological properties of HRT when using HRT for risk stratification, especially in elderly patients. PMID- 15807393 TI - Tei index as a method of evaluating left ventricular diastolic dysfunction in acute myocardial infarction. AB - INTRODUCTION: The Doppler index of overall left ventricular (LV) myocardial performance--the Tei index--has been shown to be a reliable indicator of all changes in LV systolic dysfunction, retaining an inverse relationship with the ejection fraction. The aim of this study was to examine the corresponding behaviour in relation to LV diastolic dysfunction in patients with acute myocardial infarction (AMI), a relationship that has not been studied previously. METHODS: The study included 105 patients (77 men) with first AMI who were classified into four groups according to the severity of LV diastolic dysfunction: a) 25 patients with normal diastolic function (NDF), b) 36 with decreased peak filling rate pattern (DFR), c) 33 with impaired relaxation (IR) and d) 11 with pseudonormal or restrictive physiology (PN/RP). A complete echocardiographic study, including all conventional systolic and diastolic echo/Doppler parameters as well as measurement of the Tei index, was performed on the eighth post-infarction day (mean 8.07 +/- 0.96 days) in all patients. RESULTS: In the patients with IR (0.77 +/- 0.05) the index was significantly greater than in those of the NDF (0.55 +/- 0.03, p<0.01) or DFR (0.65 +/- 0.02, p<0.01) groups. The index in the DFR group was greater than in the NDF group, though not significantly so. In contrast, the index in the PN/RP patients (0.59 +/- 0.05) was significantly lower than in the patients with IR (p<0.01), whereas it did not differ from that of the patients in the NDF or DFR groups ("pseudonormalisation" of the index). CONCLUSIONS: The Tei index detects with reliability milder types of diastolic dysfunction. However, because of its "pseudonormalisation" in patients with PN/RP, the Tei index cannot be considered a reliable indicator of more severe patterns of LV diastolic dysfunction in AMI patients. PMID- 15807394 TI - Myocardial performance index or Tei index: valuable in research but doubtful in clinical practice. PMID- 15807395 TI - Comparison of enoxaparin and unfractionated heparin in coronary angioplasty. AB - INTRODUCTION: Recent studies have shown that enoxaparin may be equally as safe and effective as unfractionated heparin during a coronary angioplasty procedure. The aim of this study was to investigate whether enoxaparin can be used effectively and safely in place of unfractionated heparin in patients undergoing emergency or programmed coronary angioplasty, regardless of the use of platelet glycoprotein IIb/IIIa inhibitors. METHODS: We compared two series of consecutive patients, who received unfractionated heparin (n = 217) or enoxaparin (n = 116) during emergency or programmed angioplasty, regardless of age, weight, renal function and the coadministration of platelet glycoprotein IIb/IIIa inhibitors. In the patients who received enoxaparin the arterial sheaths were removed immediately after the procedure. RESULTS: There were no significant differences between the two groups as regards clinical characteristics or risk factors for coronary artery disease. During a 30-day follow up no major adverse cardiac events were observed (death, myocardial infarction, target vessel revascularisation). Multivariate logistic regression analysis showed no correlation between the anticoagulant used and the occurrence of major cardiac events in the two groups of patients (log odds ratio = -9.46, p = 0.89), after controlling for age, sex, administration of platelet glycoprotein IIb/IIIa inhibitors, number of coronary lesions, number of stents used, clinical picture and risk factors for coronary artery disease. As regards the development of haematoma in the groin, the only significant independent predictive factor for this was the coadministration of platelet glycoprotein IIb/IIIa inhibitors. CONCLUSIONS: The use of enoxaparin in coronary angioplasty is safe, effective and allows faster removal of sheaths and mobilisation of the patient. PMID- 15807396 TI - The Tei index of myocardial performance: applications in cardiology. PMID- 15807397 TI - Homocysteine: a risk factor for coronary artery disease? PMID- 15807398 TI - Study of myocardial viability using gadolinium-enhanced magnetic resonance imaging. PMID- 15807399 TI - Cardiovascular magnetic resonance evaluation of patients with transposition of the great arteries following atrial switch surgical correction. AB - We present a method of imaging patients with surgically corrected transposition of the great arteries using cardiac magnetic resonance imaging and we describe the findings from a typical case. Cardiac magnetic resonance is superior to other imaging methods for patients with complex congenital heart diseases, both for establishing the initial diagnosis and for subsequent follow-up after surgical therapy. PMID- 15807400 TI - Electroanatomical mapping and ablation of upper loop reentry atrial flutter. AB - Upper loop reentry is a rare type of non isthmus-dependent atrial flutter with a reentry circuit around the superior vena cava. We present a case of atrial flutter of this type in a 75-year-old man with concomitant sick sinus syndrome. The diagnostic approach used three-dimensional electroanatomical mapping with the upgraded ENSITE 3000 system. This is the first case to be described where, apart from non-contact mapping, the circuit was also tracked in detail with three dimensional contact mapping. The reentry circuit followed a clockwise course around the superior vena cava and passed through a conduction gap in the crista terminalis. Radiofrequency ablation of this region successfully eliminated the tachycardia. PMID- 15807401 TI - Acute myocardial infarction in a patient with normal coronary arteries after an allergic reaction. AB - The case is described of a man aged 51 years who, one hour after a coronary angiographic examination that showed coronary arteries without lesions, suffered an acute inferior myocardial infarction as the result of an acute allergic reaction, probably to the iodinated contrast agent that was used. Acute myocardial infarction following an allergy is an extremely rare clinical entity that, according to existing data, is due to the local release of vasoconstrictive substances as a result of the allergic reaction. PMID- 15807402 TI - Res Ipsa Loquitur: a step along the road to liability without fault. AB - Traditionally, courts have required that the plaintiff prove the negligence of medical defendants through expert testimony. Res ipsa loquitur, on the other hand, allows juries to infer negligence from the circumstances surrounding the injury. The tension between those two propositions is obvious. It is incongruous to allow a lay jury to infer a proposition that generally demands expert proof. PMID- 15807403 TI - Prevalence and risk factors of occupational hand dermatoses in electronics workers. AB - The electronics industry is becoming an important mainstream in the workforce in some developed countries and in Taiwan. Among patients with occupational hand dermatitis in northern Taiwan, workers from electronics industries were one of the most important groups. We conducted a field investigation to determine the prevalence, patterns and risk factors of occupational hand dermatoses among electronics workers. The survey was conducted in five electronics plants using a self-administered questionnaire on skin symptoms and risk factors. Skin examination and patch testing were followed for those with symptoms compatible with hand dermatitis. A total of 3070 workers completed the questionnaire. Among them, 302 (9.8%) reported to have symptoms (itching and with either redness/scaling) compatible with contact dermatitis on hands. Hand dermatitis was associated with working in the fabrication unit and personal history of atopy and metal allergy, as well as the following job titles: wafer bonding, cutting, printing/photomasking, softening/degluing, impregnation and tin plating. Among those with reported hand dermatitis, 183 completed skin examination and patch testing, 65/183 (35.5%) were diagnosed as having irritant contact dermatitis (ICD) and 7/183 (3.8%) allergic contact dermatitis. The most important allergens were nickel, cobalt and phenylenediamine. In conclusion, Taiwanese electronics workers have a high risk of having hand dermatitis, especially ICD. Preventive efforts should be focused on the workers with risk factors or at certain worksites. PMID- 15807404 TI - Species difference in the disposition of acrylonitrile: quantitative whole-body autoradiographic study in rats and mice. AB - Previous studies from this and other laboratories have indicated the role of species difference in acrylonitrile (VCN) toxicity and its metabolism to cyanide. Our recent studies also indicated a more pronounced elimination of VCN following oral as compared to i.v. administration. To further characterize the mechanism of these differences on the distribution of VCN, quantitative whole-body autoradiographic distribution and elimination studies were conducted at various time points (0.08, 8, 24, 48 h) following the administration of an equivalent i.v. dose of 2-[14C]-VCN to male Fischer rats and male CD-1 mice. Whole-body autoradiographs obtained from freeze-dried and acid-extracted sections of rats and mice demonstrated a rapid uptake of 14C in liver, lungs, spleen and bone marrow at early time intervals. Quantitatively, the uptake, retention and covalent interaction of 14C were higher in organs of rats as compared to mice, over 48 h. Mice eliminated 74% of the total administered dose of 2-[14C]-VCN (expired air 4%, urine 16% and feces 54%), while rats eliminated only 26% of the dose (expired air 2%, urine 4% and feces 20%). Species differences in VCN toxicity seem to be correlated with its rate of elimination. The distribution and elimination data demonstrated that mice divest VCN more rapidly than rats. The study also demonstrated that administration of VCN in rats resulted in covalent interactions and retention of 2-[14C]-VCN/metabolites in the tissues thus exerting more chronic toxicity to rats than to mice. PMID- 15807405 TI - Comparing study of the effect of nanosized silicon dioxide and microsized silicon dioxide on fibrogenesis in rats. AB - This study compares the effect of nanosized silicon dioxide (nanosized SiO2) and microsized silicon dioxide (microsized SiO2) particles on fibrogenesis in rats. Wistar rats were instilled intratracheally with saline, 20 mg of nanosized SiO2 or 20 mg of microsized SiO2, and were sacrificed at 1 and 2 months after instillation. The lungs of rats were analysed for the changes of lung/body coefficient and hydroxyproline content. Changes in pathology and fibrotic grade were observed by use of hematoxylin and eosin and Van Gieson dyeing. The expression of interleukin-4 (IL-4) and transforming growth factor-beta1 (TGF beta1) was observed by use of immunohistochemical technique, and protein expression quantitatively analysed by image analysis. The lung/body coefficient and hydroxyproline content of nanosized SiO2 groups were significantly lower than those of microsized SiO2 groups at both 1 and 2 months after instillation (P<0.05 or P<0.01), but without significant differences from those of saline control groups. At 1 month after instillation, there were mainly cellular nodules (Stage I) in nanosized SiO2 group, while in microsized SiO2 group Stage II, II+ of silicotic nodules were observed. At 2 months after instillation, there were still Stage I of silicotic nodules in nanosized SiO2 group. In microsized SiO2 group mainly Stage II+, III of silicotic nodules were found. Quantity image analysis showed that the expressions of IL-4 and TGF-beta1 in nanosized SiO2 groups were significantly lower than those in microsized SiO2 groups (P<0.01), but without significant difference from those of saline control groups. Our experiment revealed that the effect of fibrogenesis of nanosized SiO2 might be milder than that of microsized SiO2 in rats, potentially resulting from nanoparticals tending to be diffused and easily translocated due to their ultrafine particle size compared to microsized particles. PMID- 15807406 TI - A dynamic simulation of bisphenol A dosimetry in neuroendocrine organs. AB - Bisphenol A (BPA) is a known xenoestrogen with similar properties to 17beta estradiol. BPA and estrogen are hydrophobic compounds, and this affects the pharmacokinetics of both compounds in mammals. In a previous study we measured the distribution of BPA in female F344 rats exposed to oral doses of 0.1, 10 and 100 mg/kg. The results showed distribution to target neuroendocrine organs at all doses tested. Using these results, we developed a pharmacokinetic model to predict the dynamic uptake and excretion of BPA by various routes of exposure (po, iv, sc, ip). The model was able to simulate the entire time course (48 h) following various routes of exposure in rats over the dose ranges tested. The model indicated that the ultimate tissue uptake of BPA was established by the rapid initial transfer of free BPA into tissues. After free BPA enters the systemic circulation, metabolism and excretion reactions cause a relatively short duration and rapid decline. This period is followed by a slower long-term decline characteristic of BPA's biphasic pharmacokinetics. Plasma protein and tissue binding reactions established the long-term half-life of BPA in the body. Route differences in tissue uptake were directly related to the competition between transfer and binding reactions during the absorption phase. PMID- 15807407 TI - Distribution of bisphenol A in the neuroendocrine organs of female rats. AB - The distribution of 14C-bisphenol A (BPA) in plasma and neuroendocrine organs was determined in Fischer 344 female rats following three oral doses (0.1, 10 or 100mg/kg). Plasma and tissue maximum concentrations (Cmax) were reached within 15 30 min of dosing. Plasma areas-under-the-curve (AUC) ranged from 0.06 to 53.9 microg-h/mL. The AUCs of the pituitary gland and uterus/gonads were 16-21% higher than that of plasma. The AUCs of hypothalamus and the rest of the brain were 43.7% and 77% of the plasma AUCs, respectively. In the brain tissue, the exposure increased linearly with the oral dose, as the dose was increased from 0.1 to 10 and 100 mg/kg; the exposure in the brain relative to the plasma increased by factors of 1, 1.19 and 1.24. This indicates that the brain barrier systems do not limit the access of the lipophilic BPA to the brain. The increases of the uterus/gonads relative to the plasma were 1, 1.07 and 1.04. Tissue partitioning was also examined in vitro by the uptake of 14C-BPA. The BPA tissue/blood partition coefficients were as follows: heart, 7.5; liver, 6.1; kidney, 6.4; fat, 3.6; muscle, 2.6; breast, 3.6; ovaries, 9.1; uterus, 5.9; stomach, 5.1; and small intestine, 6.7. The tissue/cerebrospinal fluid partition coefficients were as follows: pituitary gland, 12.8; brain stem, 6.1; cerebellum, 6.4; hippocampus, 7.1; hypothalamus, 6.1; frontal cortex, 4.9; and caudate nucleus, 6.8. PMID- 15807408 TI - Markers of cadmium exposure in workers in a cadmium pigment factory after changes in the exposure conditions. AB - The objective of this study was to assess changes in concentrations of cadmium in the blood (Cd-B), cadmium in the urine (Cd-U), beta2-microglobulin in the serum (beta2-mG-S) and beta2-microglobulin in the urine (beta2-mG-U) of workers at a cadmium (Cd) pigment factory in Japan in which exposure conditions improved. We evaluated reversibility of these markers in continuously employed workers in relation to changes in exposure levels resulting from improvements in the workplace and the reduced production of Cd. Our study involved both environmental and biological monitoring. Data were collected for four years. We measured the Cd concentration in the air of each work area, using the time-weighted average (TWA). Cd-B and Cd-U were measured in workers as direct indices of Cd exposure. beta2-mG-S and beta2-mG-U were measured as markers of renal tubular function. Exposure levels were high in all work areas, according to the criteria set by the American Conference of Governmental Industrial Hygienists (ACGIH). Workers' Cd-B and Cd-U concentrations reflected high levels of exposure. Correlation was found between these direct indices and beta2-mG-S concentrations. Since the second year, ambient Cd concentrations decreased and reacted markers have been improved. Our results suggest that Cd-B, Cd-U, beta2-mG-S and beta2-mG-U are appropriate markers for monitoring both the level of Cd exposure and the tubular function of workers. Reversibility of urinary low molecular weight protein was observed in the workers over the four years. PMID- 15807409 TI - Hormesis effect of trace metals on cultured normal and immortal human mammary cells. AB - An in vitro study was conducted to determine the effects of variable concentrations of trace metals on human cultured mammary cells. Monolayers of human mortal (MCF-12A) and immortal (MDA-MB231) mammary epithelial cells were incubated in the absence or presence of increasing concentrations of arsenic (As), mercury (Hg) and copper (Cu) for 24-h, 72-h, 4-d, and 7-d. The MTT assay was used to assess viability for all time periods and cell proliferation was monitored for 4-d and 7-d studies. Monolayers were also labeled with rhodamine 110 (R-6501), Sytox green, and Celltiter blue fluorescent dyes as indicators for intracellular esterase activity, nucleic acid staining, and cell reduction/viability, respectively. Total incubation time with chemical plus dyes was 24 h. For 24-h and 72-h studies, cells were seeded in 96-well plates, after which confluent monolayers were exposed to increasing concentrations of chemicals. For 4-d and 7-d studies, cells were seeded in 12-well plates at 1/3 confluent density (day 0) and exposed to increasing concentrations of metals on day 1. All cells were counted on days 4 and 7. In addition, test medium was removed from select groups of cultures on day 4, replaced with fresh medium in the absence of chemical (recovery studies), and assays were performed on day 7 as above. The data suggest that there is a consistent protective and/or stimulating effect of metals at the lowest concentrations in MCF-12A cells that is not observed in immortal MDA-MB231 cells. In fact, cell viability of MCF-12A cells is stimulated by otherwise equivalent inhibitory concentrations of As, Cu, and Hg on MDA-MB231 cells at 24-h. Whereas As and Hg suppress proliferation and viability in both cell lines after 4-d and 7-d of exposure, Cu enhances cell proliferation and viability of MCF-12A cells. MDA-MB231, however, recover better after 4-days of toxic insult. In addition, nutritional manipulation of media between the cell lines, or pretreatment with penicillamine, did not alter the hormesis effect displayed by MCF-12A. Growth of these cells however was not maintained in the alternative medium. The study demonstrates that a hormesis effect from trace metals is detectable in cultured mammary cells; fluorescent indicators, however, are not as sensitive as cell proliferation or MTT in recognizing the subtle responses. Also, sensitivity of mammary cells to lower concentrations of Cu, a biologically important trace metal, may play an important role in controlling cellular processes and proliferation. The ability to detect this in vitro phenomenon implies that similar processes, occurring in vivo, may be responsible for the development, induction, or enhancement of human cancers. PMID- 15807410 TI - Metal mixture inhalation (Cd-Pb) and its effects on the bronchiolar epithelium. An ultrastructural approach. AB - The current study explores the effects of the inhalation of lead (Pb), Cd and its mixture (Pb-Cd) in a mice model, analysing metal concentrations in the lung, and the morphological modifications in the bronchiolar epithelium identified by scanning electron microscopy after eight weeks of inhalation. Our results indicate that metal concentrations in lung were higher compared to controls; however, Pb concentrations drastically decrease in the mixture. This reduction was also observed in the inhalation chamber. The main changes observed in the bronchiole were mostly in the mixture. The modifications were mainly given by Cd alone and in the mixture, with a decreased number of nonciliated bronchiolar cells and an increased number of bundles of dividing cells. The additive effect of Pb-Cd is suggested, as the extensive damage observed was more evident when mice were exposed to the mixture, and the results endured more research in the area of inhaled mixtures. PMID- 15807411 TI - Gene-expression profiling using suppression-subtractive hybridization and cDNA microarray in rat mononuclear cells in response to welding-fume exposure. AB - Welders with radiographic pneumoconiosis abnormalities have shown a gradual clearing of the X-ray identified effects following removal from exposure. In some cases, the pulmonary fibrosis associated with welding fumes appears in a more severe form in welders. Accordingly, for the early detection of welding-fume exposure-induced pulmonary fibrosis, the gene expression profiles of peripheral mononuclear cells from rats exposed to welding fumes were studied using suppression-subtractive hybridization (SSH) and a cDNA microarray. As such, Sprague-Dawley rats were exposed to a stainless steel arc welding fume for 2 h/day in an inhalation chamber with a 1107.5 +/- 2.6 mg/m3 concentration of total suspended particulate (TSP) for 30 days. Thereafter, the total RNA was extracted from the peripheral blood mononuclear cells, the cDNA synthesized from the total RNA using the SMART PCR cDNA method, and SSH performed to select the welding-fume exposure-regulated genes. The cDNAs identified by the SSH were then cloned into a plasmid miniprep, sequenced and the sequences analysed using the NCBI BLAST programme. In the SSH cloned cDNA microarray analysis, five genes were found to increase their expression by 1.9-fold or more, including Rgs 14, which plays an important function in cellular signal transduction pathways; meanwhile 36 genes remained the same and 30 genes decreased their expression by more than 59%, including genes associated with the immune response, transcription factors and tyrosine kinases. Among the 5200 genes analysed, 256 genes (5.1%) were found to increase their gene expression, while 742 genes (15%) decreased their gene expression in response to the welding-fume exposure when tested using a commercial 5.0k DNA microarray. Therefore, unlike exposure to other toxic substances, prolonged welding-fume exposure was found to substantially downregulate many genes. PMID- 15807412 TI - Detecting the historical signature of key innovations using stochastic models of character evolution and cladogenesis. AB - Phylogenetic evidence for biological traits that increase the net diversification rate of lineages (key innovations) is most commonly drawn from comparisons of clade size. This can work well for ancient, unreversed traits and for correlating multiple trait origins with higher diversification rates, but it is less suitable for unique events, recently evolved innovations, and traits that exhibit homoplasy. Here I present a new method for detecting the phylogenetic signature of key innovations that tests whether the evolutionary history of the candidate trait is associated with shorter waiting times between cladogenesis events. The method employs stochastic models of character evolution and cladogenesis and integrates well into a Bayesian framework in which uncertainty in historical inferences (such as phylogenetic relationships) is allowed. Applied to a well known example in plants, nectar spurs in columbines, the method gives much stronger support to the key innovation hypothesis than previous tests. PMID- 15807413 TI - The contribution of spontaneous mutation to variation in environmental responses of Arabidopsis thaliana: responses to light. AB - It has been hypothesized that new, spontaneous mutations tend to reduce fitness more severely in more stressful environments. To address this hypothesis, we grew plants representing 20 Arabidopsis thaliana mutation-accumulation (M-A) lines, advanced to generation 17, and their progenitor, in differing light conditions. The experiment was conducted in a greenhouse, and two treatments were used: full sun and shade, in which influx of red light was reduced relative to far-red. The shade treatment was considered the more stressful because mean absolute fitness was lower in that treatment, though not significantly so. Plants from generation 17 of M-A developed significantly faster than those from generation 0 in both treatments. A significant interaction between generation and treatment revealed that, counter to the hypothesis, M-A lines tended to have higher fitness on average relative to the progenitor in the shaded conditions, whereas, in full sun, the two generations were similar in fitness. A secondary objective of this experiment was to characterize the contribution of new mutations to genotype x environment interaction. We did not, however, detect a significant interaction between M-A line and treatment. Plots of the line-specific environmental responses indicate no tendency of new mutations to contribute to fitness trade offs, between environments. They also do not support a model of conditionally deleterious mutation, in which a mutant reduces fitness only in a particular environment. These results suggest that interactions between genotype and light environment previously documented for A. thaliana are not explicable primarily as a consequence of steady input of spontaneous mutations having environment specific effects. PMID- 15807414 TI - Multigenerational effects of inbreeding in Cucurbita pepo ssp. texana (Cucurbitaceae). AB - The shape of the fitness function relating the decline in fitness with coefficient of inbreeding (f) can provide evidence concerning the genetic basis of inbreeding depression, but few studies have examined inbreeding depression across a range of f using noncultivated species. Futhermore, studies have rarely examined the effects of inbreeding depression in the maternal parent on offspring fitness. To estimate the shape of the fitness function, we examined the relationship between f and fitness across a range off from 0.000 to 0.875 for components of both male and female fitness in Cucurbita pepo ssp. texana. Each measure of female fitness declined with f, including pistillate flower number, fruit number, seed number per fruit, seed mass per fruit, and percentage seed germination. Several aspects of male fitness also declined with f, including staminate flower number, pollen number per flower, and the number of days of flowering, although cumulative inbreeding depression was less severe for male (0.34) than for female function (0.39). Fitness tended to decline linearly with f between f = 0.00 and f = 0.75 for most traits and across cumulative lifetime fitness (mean = 0.66), suggesting that individual genes causing inbreeding depression are additive and the result of many alleles of small effect. However, most traits also showed a small reduction in inbreeding depression between f = 0.75 and f = 0.875, and evidence of purging or diminishing epistasis was found for in vitro pollen-tube growth rate. To examine inbreeding depression as a maternal effect, we performed outcross pollinations on f = 0.0 and f = 0.5 mothers and found that depression due to maternal inbreeding was 0.07, compared to 0.10 for offspring produced through one generation of selfing. In at least some families, maternal inbreeding reduced fruit number, seed number and mass, staminate flower number, pollen diameter, and pollen-tube growth rate. Collectively these results suggest that, while the fitness function appears to be largely linear for most traits, maternal effects may compound the effects of inbreeding depression in multigenerational studies, though this may be partially offset by purging or diminishing epistasis. PMID- 15807415 TI - Offspring sex ratio under inbreeding and outbreeding in a gynodioecious plant. AB - Silene vulgaris is a gynodioecious plant native to Eurasia and now found throughout much of North America. Using hermaphrodite plants from three geographic regions (Stamford, NY; Broadway,VA; and Giles Co., VA) and four local populations within each region, we employed a hierarchical crossing design to explore the geographic structure of sex determining genes. Sex determination in this species is cytonuclear involving multiple cytoplasmic male sterility and nuclear restorer loci. Due to dominance effects within nuclear restorer loci, self-fertilization of hermaphrodites heterozygous at restorer loci should produce some homozygous recessive female offspring. Female offspring may also result from outcrossing among related individuals. At greater geographic and genetic distances, mismatches between cytoplasmic and nuclear sex determining genes should also produce high frequencies of female offspring if coevolution between cytoplasmic and nuclear sex determining alleles occurs independently among widely separated populations. We found evidence of dominance effects among nuclear restorer loci but no evidence of nuclear-cytoplasmic mismatches at the regional level. Of 63 maternal lines, 55 produced at least one female offspring when self fertilized. Outcrossing within populations produced significantly fewer female offspring than self-fertilization. Outcrossing among regions produced the lowest proportion of female offspring, significantly fewer than outcrossing among populations within regions. Regions responded differently to among-region outcrossing with pollen donors from the two Virginia regions producing far fewer female offspring with New York dams than crosses among New York populations. These results indicate that nuclear restoration is complex, involving multiple loci with epistatic interactions and that most hermaphrodites in nature are heterozygous at one or more restorer locus. Further, regional differences in restorer frequencies indicate significant genetic structure for sex determining genes at large geographic scales, perhaps reflecting invasion history. PMID- 15807416 TI - Dispersal in a parasitic worm and its two hosts: consequence for local adaptation. AB - Characterizing host and parasite population genetic structure and estimating gene flow among populations is essential for understanding coevolutionary interactions between hosts and parasites. We examined the population genetic structure of the trematode Schistosoma mansoni and its two host species (the definitive host Rattus rattus and the intermediate host Biomphalaria glabrata) using microsatellite markers. Parasites were sampled from rats. The study was conducted in five sites of the Guadeloupe Island, Lesser Antilles. Mollusks display a pattern of isolation by distance whereas such a pattern is not found neither in schistosomes nor in rats. The comparison of the distribution of genetic variability in S. mansoni and its two host species strongly suggests that migration of parasites is principally determined by that of the vertebrate host in the marshy focus of Guadeloupe. However, the comparison between genetic differentiation values in schistosomes and rats suggests that the efficacy of the schistosome rat-mediated dispersal between transmission sites is lower than expected given the prevalence, parasitic load and migration rate of rats among sites. This could notably suggest that rat migration rate could be negatively correlated to the age or the infection status of individuals. Models made about the evolution of local adaptation in function of the dispersal rates of hosts and parasites suggest that rats and mollusks should be locally adapted to their parasites. PMID- 15807417 TI - Cryptic speciation and host-race formation in a purportedly generalist tumbling flower beetle. AB - Host-race formation remains controversial as a source of herbivorous insect diversity, and examples of host races are still fairly scarce. In this study, analysis of five enzyme loci in the ostensibly generalist tumbling flower beetle Mordellistena convicta (Coleoptera: Mordellidae) revealed hidden host-plant and plant-organ related genetic differentiation. Mordellistena convicta turned out to be a complex of cryptomorphic species, each with fewer hosts than the nominal species. These cryptic species, in turn, were divided into taxa that showed host race characteristics: samples from different host plants and organs exhibited (1) genetic indications of partial reproductive isolation, (2) differences in size and emergence timing that suggested divergent host-related selection, and (3) among-host selective differences in mortality from parasitoids. Host-race formation in M. convicta, which has a somewhat different life history from the well-studied host races, enlarges the group of insects considered likely to undergo this process. The widespread sympatry of the M. convicta species complex, along with its spectrum of host-correlated genetic differentiation, suggests that these host specialist taxa developed in sympatry. PMID- 15807418 TI - Outbreeding depression, but no inbreeding depression in haplodiploid Ambrosia beetles with regular sibling mating. AB - In sexual reproduction the genetic similarity or dissimilarity between mates strongly affects offspring fitness. When mating partners are too closely related, increased homozygosity generally causes inbreeding depression, whereas crossing between too distantly related individuals may disrupt local adaptations or coadaptations within the genome and result in outbreeding depression. The optimal degree of inbreeding or outbreeding depends on population structure. A long history of inbreeding is expected to reduce inbreeding depression due to purging of deleterious alleles, and to promote outbreeding depression because of increased genetic variation between lineages. Ambrosia beetles (Xyleborini) are bark beetles with haplodiploid sex determination, strong local mate competition due to regular sibling mating within the natal chamber, and heavily biased sex ratios. We experimentally mated females of Xylosandrus germanus to brothers and unrelated males and measured offspring fitness. Inbred matings did not produce offspring with reduced fitness in any of the examined life-history traits. In contrast, outcrossed offspring suffered from reduced hatching rates. Reduction in inbreeding depression is usually attributed to purging of deleterious alleles, and the absence of inbreeding depression in X. germanus may represent the highest degree of purging of all examined species so far. Outbreeding depression within the same population has previously only been reported from plants. The causes and consequences of our findings are discussed with respect to mating strategies, sex ratios, and speciation in this unusual system. PMID- 15807419 TI - Multiple cases of striking genetic similarity between alternate electric fish signal morphs in sympatry. AB - Striking trait polymorphisms are worthy of study in natural populations because they can often shed light on processes of phenotypic divergence and specialization, adaptive evolution, and (in some cases) the early stages of speciation. We examined patterns of genetic variation within and between populations of mormyrid fishes that are morphologically cryptic in sympatry but produce alternate types of electric organ discharge (EOD). Other species in a large group containing a clade of these morphologically cryptic EOD types produce stereotyped, species-typical EOD waveforms thought to function in mate recognition. First, for six populations from Gabon's Brienomyrus species flock, we confirm that forms of electric fish that exhibit distinctive morphologies and unique EOD waveforms (i.e., good reference species) are reproductively isolated from coexisting congeners. These sympatric species deviate from genetic panmixia across five microsatellite loci. Given this result, we examined three focal pairs of syntopic and morphologically cryptic EOD waveform types that are notable exceptions to the pattern of robust genetic partitioning among unique waveform classes within assemblages. These exceptional pairs constitute a monophyletic group within the Brienomyrus flock known as the magnostipes complex. One member of each pair (type I) produces a head-negative EOD, while the other member (either type II or type III, depending on location) produces a longer duration EOD differing in waveform from type I. We show that signal development in these pairs begins with juveniles of all magnostipes-complex morphs emitting head positive EODs resembling those of type II adults. Divergence of EOD waveforms occurs with growth such that there are two discrete and fixed signal types in morphologically indistinguishable adults at each of several localities. Strong microsatellite partitioning between allopatric samples of any of these morphologically cryptic signal types suggests that geographically isolated populations are genetically decoupled from one another. By contrast, sympatric morphs appear genetically identical across microsatellite loci in Mouvanga Creek and the Okano River and only very weakly diverged, if at all, in the Ivindo River. Our results for the magnostipes complex fail to detect species boundaries between the focal morphs and are, instead, fully consistent with the existence of relatively stable signal dimorphisms at each of several different localities. No mechanism for the maintenance of this electrical polymorphism is suggested by the known natural history of the magnostipes complex. Despite a lack of evidence for genetic differentiation, the possibility of incipient sympatric speciation between morphs (especially type I and type II within the Ivindo River) merits further testing due to behavioral and neurobiological lines of evidence implying a general role for stereotyped EOD waveforms in species recognition. We discuss alternative hypotheses concerning the origins, stability, and evolutionary significance of these intriguing electrical morphs in light of geographical patterns of population structure and signal variation. PMID- 15807420 TI - Contrasting quaternary histories in an ecologically divergent sister pair of low dispersing intertidal fish (Xiphister) revealed by multilocus DNA analysis. AB - Recurrent glacial advances have shaped community histories across the planet. While biogeographic responses to glaciations likely varied with latitude, the consequences for temperate marine communities histories are less clear. By coalescent analyses of multiloci DNA sequence data (mitochondrial DNA control region, alpha-enolase intron, and alpha-tropomyosin intron) collected from a low dispersing sister pair of rocky intertidal fishes commonly found from southeastern Alaska to California (Xiphister atropurpureus and X. mucosus), we uncover two very different responses to historical glaciations. A variety of methods that include a simulation analysis, coestimates of migration and divergence times, and estimates of minimum ages of populations sampled up and down the North American Pacific coast all strongly revealed a history of range persistence in X. atropurpureus and extreme range contraction and expansion from a southern refugium in X. mucosus. Furthermore, these conclusions are not sensitive to the independent estimates of the DNA substitution rates we obtain. While gene flow and dispersal are low in both species, the widely different histories are rather likely to have arisen from ecological differences such as diet breadth, generation time, and habitat specificity. PMID- 15807421 TI - Parallel evolution of sexual isolation in sticklebacks. AB - Mechanisms of speciation are not well understood, despite decades of study. Recent work has focused on how natural and sexual selection cause sexual isolation. Here, we investigate the roles of divergent natural and sexual selection in the evolution of sexual isolation between sympatric species of threespine sticklebacks. We test the importance of morphological and behavioral traits in conferring sexual isolation and examine to what extent these traits have diverged in parallel between multiple, independently evolved species pairs. We use the patterns of evolution in ecological and mating traits to infer the likely nature of selection on sexual isolation. Strong parallel evolution implicates ecologically based divergent natural and/or sexual selection, whereas arbitrary directionality implicates nonecological sexual selection or drift. In multiple pairs we find that sexual isolation arises in the same way: assortative mating on body size and asymmetric isolation due to male nuptial color. Body size and color have diverged in a strongly parallel manner, similar to ecological traits. The data implicate ecologically based divergent natural and sexual selection as engines of speciation in this group. PMID- 15807422 TI - Marine radiations at small geographic scales: speciation in neotropical reef gobies (Elacatinus). AB - Studies of speciation in the marine environment have historically compared broad scale distributions and estimated larval dispersal potential to infer the geographic barriers responsible for allopatric speciation. However, many marine clades show high species diversity in geographically restricted areas where barriers are not obvious and estimated dispersal potential should bring many sister taxa into contact. Genetic differentiation at small (separation < 1000 km) spatial scales could facilitate speciation by mechanisms other than the gradual accumulation of reproductive isolation during extended allopatry, such as ecological adaptation to local environmental conditions or the rapid evolution of genes tied to mate recognition, but the role of each of these possibilities has not been simultaneously explored for any species-rich marine taxon. Here, we develop a robust phylogenetic framework for 31 taxa from a species-rich group of Neotropical reef fishes (Gobiidae: Elacatinus) using 3230 bp from one mitochondrial and two nuclear gene regions. We use this framework to explore the contribution of large- and small-scale geographic isolation, ecological differentiation, and coloration toward the formation and maintenance of species. Although species of Elacatinus occur on both sides of the Isthmus of Panama, no sister species are separated by this barrier. Instead, our results indicate that sister taxa occur within oceans. Sister taxa usually differ by coloration, and more distantly related sympatric species frequently differ by resource use. This suggests that some combination of coloration and ecological differences may facilitate assortative mating at range boundaries or in sympatry. Overall, speciation in Elacatinus is consistent with a model of recurring adaptive radiations in stages taking place at small geographic scales. PMID- 15807423 TI - Phylogeography and demography of sympatric sister surfperch species, Embiotoca jacksoni and E. lateralis along the California coast: historical versus ecological factors. AB - With 18 closely related endemic species that radiated in a diversity of ecological niches, the California surfperches (Embiotocidae) species flock is a good candidate for the study of sympatric speciation. Resource partitioning has been suggested as an important driving force in the radiation of the surfperch family. Within the family, two congeneric sister species, Embiotoca jacksoni and E. lateralis, are known to compete strongly for a preferred single food resource and may be used as a model of ecological interactions for the family. Along the California coast, the distribution of the two species differs. Embiotoca jacksoni has a continuous range, whereas E. lateralis shows a disjunction with a distribution gap in the Southern California Bight. Two hypotheses may explain this disjunct distribution. Ecological competition may have displaced E. lateralis in favor of E. jacksoni. Alternatively, a common vicariant event may have separated the species into northern and southern populations, followed by secondary contact in E. jacksoni but not in E. lateralis. The two hypotheses predict different phylogeographic and demographic signatures. Using a combined phylogeographic and coalescent approach based on mitochondrial control region data, we show that vicariance can only account for a portion of the observed divergences. Our results are compatible with a significant role played by ecological competition in the southern range of the species. PMID- 15807424 TI - Advertisement-call preferences in diploid-tetraploid treefrogs (Hyla chrysoscelis and Hyla versicolor): implications for mate choice and the evolution of communication systems. AB - Signals used for mate choice and receiver preferences are often assumed to coevolve in a lock-step fashion. However, sender-receiver coevolution can also be nonparallel: even if species differences in signals are mainly quantitative, females of some closely related species have qualitatively different preferences and underlying mechanisms. Two-alternative playback experiments using synthetic calls that differed in fine-scale temporal properties identified the receiver criteria in females of the treefrog Hyla chrysoscelis for comparison with female criteria in a cryptic tetraploid species (H. versicolor); detailed preference functions were also generated for both species based on natural patterns of variation in temporal properties. The species were similar in three respects: (1) pulses of constant frequency were as attractive as the frequency-modulated pulses typical of conspecific calls; (2) changes in preferences with temperature paralleled temperature-dependent changes in male calls; and (3) preference functions were unimodal, with weakly defined peaks estimated at values slightly higher than the estimated means in conspecific calls. There were also species differences: (1) preference function slopes were steeper in H. chrysoscelis than in H. versicolor; (2) preferences were more intensity independent in H. chrysoscelis than in H. versicolor; (3) a synergistic effect of differences in pulse rate and shape on preference strength occurred in H. versicolor but not in H. chrysoscelis; and (4) a preference for the pulse shape typical of conspecific calls was expressed at the species-typical pulse duration in H. versicolor but not in H. chrysoscelis. However, females of H. chrysoscelis did express a preference based on pulse shape when tested with longer-than-average pulses, suggesting a hypothesis that could account for some examples of nonparallel coevolution. Namely, preferences can be hidden or revealed depending on the direction of quantitative change in a signal property relative to the threshold for resolving differences in that property. The results of the experiments reported here also predict patterns of mate choice within and between contemporary populations. First, intraspecific mate choice in both species is expected to be strongly influenced by variation in temperature among calling males. Second, simultaneous differences in pulse rate and pulse shape are required for effective species discrimination by females of H. versicolor but not by females of H. chrysoscelis. Third, there is greater potential for sexual selection within populations and for discrimination against calls produced by males in other geographically remote populations in H. chrysoscelis than in H. versicolor. PMID- 15807425 TI - Convergence and the multidimensional niche. AB - Convergent evolution has played an important role in the development of the ecological niche concept. We investigated patterns of convergent and divergent evolution of Caribbean Anolis lizards. These lizards diversified independently on each of the islands of the Greater Antilles, producing the same set of habitat specialists on each island. Using a phylogenetic comparative framework, we examined patterns of morphological convergence in five functionally distinct sets of morphological characters: body size, body shape, head shape, lamella number, and sexual size dimorphism. We find evidence for convergence among members of the habitat specialist types for each of these five datasets. Furthermore, the patterns of convergence differ among at least four of the five datasets; habitat specialists that are similar for one set of characters are often greatly different for another. This suggests that the habitat specialist niches into which these anoles have evolved are multidimensional, involving several distinct and independent aspects of morphology. PMID- 15807426 TI - Little effect of extrapair paternity on the opportunity for sexual selection in Savannah sparrows (Passerculus sandwichensis). AB - Extrapair paternity (EPP) can dramatically increase the opportunity for sexual selection if relatively few males are able to monopolize the majority of fertilizations in a population. Although recent work with birds suggests that EPP can increase the standardized variance in male reproductive success (Is) as much as 13-fold, only a male's within-pair success is typically quantified with any accuracy. In most cases, nearly half of all extrapair young are of unknown parentage. A strong, negative correlation across studies between the proportion of extrapair young for which parentage is known and the apparent effect of EPP on Is (r(s) = -0.71, P = 0.013, N = 13 studies) suggests that the incomplete sampling of extrapair sires has greatly exaggerated the influence of EPP. To achieve a more thorough accounting of EPP and its importance to variation in male fitness, we used a suite of four to six microsatellite loci to identify extrapair young and their sires in a polygynous population of Savannah sparrows (Passerculus sandwichensis). Pooling over the 2002 and 2003 breeding seasons, 79 of 116 females (68.1%) produced young outside of the pairbond and 194 of 411 offspring (47.2%) were extrapair. We identified sires for 96.4% of all young (N = 396), including sires for 92.3% of the extrapair young (N = 179), allowing us to partition Is into within-pair and extrapair components. In both years, EPP related fitness components generated more variation in male reproductive success than the number or quality of within-pair mates. Differences among males in the number of extrapair mates alone accounted for 56.6% of Is in 2002 and for 23.6% of Is in 2003. Nonetheless, in absolute terms, the occurrence of EPP on Kent Island increased the opportunity for sexual selection less than two-fold. Averaging over the two years, Is was only 78% higher than Is,app, the variance in male reproductive success that would have occurred had EPP been nonexistent and males sired all young on their territories. Likewise, across nine socially monogamous species, we found no correlation between the extent of EPP and its effect on the opportunity for sexual selection (Is/Is,app) and only a marginally significant positive correlation between EPP and Is itself. Taken together, our results suggest that the relationship between EPP and sexual selection in birds may be much less strong and much less straightforward than commonly thought. PMID- 15807427 TI - The effects of sex and mutation rate on adaptation in test tubes and to mouse hosts by Saccharomyces cerevisiae. AB - Some hypotheses for the evolution of sex focus on adaptation to changing or heterogeneous environments, but these hypotheses have rarely been tested. We tested for advantages of sex and of increased mutation rates in yeast strains in two contrasting environments: a standard and relatively homogeneous laboratory environment of minimal medium in test tubes, and the variable environment of a mouse brain experienced by pathogenic strains. Evolving populations were founded as equal mixtures of sexual and obligately asexual genotypes. In the sexuals, cycles of sporulation, meiosis, and mating were induced approximately every 50 mitotic generations, with the asexuals undergoing sporulation but not ploidy cycles or recombination. In both environments, replicate negative control populations established with the same pair of strains were propagated with neither mating nor meiosis. In test tubes with no sex induced, sexuals were fixed in all five replicates within 250 mitotic generations, whereas in mice with no sex induced, asexuals were fixed in all four replicates by 170 generations. Inducing sex altered these outcomes in opposite directions in test tubes and mice, decreasing the fixation frequencies of sexuals in test tubes but increasing them in mice. These contrasts with asexual controls suggest an advantage for sex in mice but not in test tubes, although there was no difference between test tubes and mice in the numbers of populations fixed-for sexuals. In analogous experiments testing for an advantage of increased mutation rates, wild-type genotypes became fixed at the expense of mutators in every replicate of both test tube and mouse populations, indicating a disadvantage for mutators in both environments. Increased rates of point mutation do not appear to accelerate adaptation. PMID- 15807428 TI - Parasite burden and constitution of major histocompatibility complex in the Malagasy mouse lemur, Microcebus murinus. AB - We investigated the importance of the major histocompatibility complex (MHC) constitution on the parasite burden of free-ranging mouse lemurs (Microcebus murinus) in four littoral forest fragments in southeastern Madagascar. Fourteen different MHC class II DRB-exon 2 alleles were found in 228 individuals with high levels of sequence divergence between alleles. More nonsynonymous than synonymous substitutions in the functional important antigen recognition and binding sites indicated selection processes maintaining MHC polymorphism. Animals from the four forest fragments differed in their infection status (being infected or not), in the number of different nematode morphotypes per individual (NNI) as well as in the fecal egg counts (FEC) values. Heterozygosity in general was uncorrelated with any of these measures of infection. However, a positive relationship was found between specific alleles and parasite load. Whereas the common allele Mimu DRB*1 was more frequently found in infected individuals and in individuals with high NNI and FEC values (high parasite load), the rare alleles Mimu-DRB*6 and 10 were more prevalent in uninfected individuals and in individuals with low NNI and FEC values (low parasite load). These three alleles associated with parasite load had unique amino acid motifs in the antigen binding sites. This distinguished them from the remaining 11 Mimu-DRB alleles. Our results support the hypothesis that MHC polymorphism in M. murinus is maintained through pathogen-driven selection acting by frequency-dependent selection. This is the first study of the association of MHC variation and parasite burden in a free-ranging primate. PMID- 15807429 TI - Selection on mothers and offspring: whose phenotype is it and does it matter? AB - Reproductive and early life-history traits can be considered aspects of either offspring or maternal phenotype, and their evolution will therefore depend on selection operating through offspring and maternal components of fitness. Furthermore, selection at these levels may be antagonistic, with optimal offspring and maternal fitness occurring at different phenotypic values. We examined selection regimes on the correlated traits of birth weight, birth date, and litter size in Soay sheep (Ovis aries) using data from a long-term study of a free-living population on the archipelago of St. Kilda, Scotland. We tested the hypothesis that selective constraints on the evolution of the multivariate phenotype arise through antagonistic selection, either acting at offspring and maternal levels, or on correlated aspects of phenotype. All three traits were found to be under selection through variance in short-term and lifetime measures of fitness. Analysis of lifetime fitness revealed strong positive directional selection on birth weight and weaker selection for increased birth date at both levels. However, there was also evidence for stabilizing selection on these traits at the maternal level, with reduced fitness at high phenotypic values indicating lower phenotypic optima for mothers than for offspring. Additionally, antagonistic selection was found on litter size. From the offspring's point of view it is better to be born a singleton, whereas maternal fitness increases with average litter size. The decreased fitness of twins is caused by their reduced birth weight; therefore, this antagonistic selection likely results from trade offs between litter size and birth weight that have different optimal resolutions with respect to offspring and maternal fitness. Our results highlight how selection regimes may vary depending on the assignment of reproductive and early life-history traits to either offspring or maternal phenotype. PMID- 15807430 TI - The file-drawer problem revisited: a general weighted method for calculating fail safe numbers in meta-analysis. AB - Quantitative literature reviews such as meta-analysis are becoming common in evolutionary biology but may be strongly affected by publication biases. Using fail-safe numbers is a quick way to estimate whether publication bias is likely to be a problem for a specific study. However, previously suggested fail-safe calculations are unweighted and are not based on the framework in which most meta analyses are performed. A general, weighted fail-safe calculation, grounded in the meta-analysis framework, applicable to both fixed- and random-effects models, is proposed. Recent meta-analyses published in Evolution are used for illustration. PMID- 15807431 TI - [Amino acid and lipid metabolism in embryos of flesh fowl with different yolk mass]. AB - It has been proposed that variations in relative yolk mass in a population of flesh fowl be used as a model of development of nidicolous and nidifugous birds. During development of the eggs with a high proportion of yolk, an excess of lipids is cleaved at a higher rate and oxidized until day 17 of incubation, while in the embryos developing from the eggs with a low relative yolk mass, amino acids are intensely cleaved during the period preceding the hatching. Significant differences in the body content of cystine were found in 17-day embryos and upon hatching, thus suggesting a delayed activity of the genes encoding keratins in the group corresponding to the seminidicolous type according to the egg content of lipids. These biochemical differences question the widespread concept on the occurrence of dichotomy by the end of embryogenesis and beginning of neonatal growth of nidifugous and nidicolous birds. PMID- 15807432 TI - [Age-related features of protein synthesis rhythm in hepatocytes. Effects of extracellular medium]. AB - Cell interactions have been studied in cultures pf hepatocytes from young and old rats. The rhythm of protein synthesis is an index of cell interaction and synchronization in culture, while the amplitude of oscillations characterized cell cooperation in an aggregate rhythm. The mean rhythm amplitude in the culture of hepatocytes from old rats is twice lower than that from young rats. Gangliosides (mixture, bovine brain gangliosides) and alpha1-adrenomimetic phenylephrine enhanced synchronization of cultures of the cells from old rats and increased the amplitude of oscillations to the level of young animals. Addition of rat blood serum (10%) to the medium revealed the rhythm of protein synthesis in the culture, asynchronous in the control, i.e., led to their synchronization. In media with young and old rat blood sera, oscillations were intense, with high amplitudes, and low, respectively. Addition of bovine brain gangliosides to a medium with old rat blood serum increased the amplitudes of oscillations to a level of the rhythm stimulated by the young rat serum. Thus, the cells of old animals can fully perceive synchronizing factors and, in the case of their increased concentration, the rhythm of protein synthesis in old animals did not differ from that in young rats. Current data on biochemical mechanisms underlying intercellular cooperation in the formation of population rhythm of protein synthesis have been discussed. PMID- 15807433 TI - [Phorbol 12-myristate 13-acetate inhibits apoptosis in erythroleukemia K562 cells induced by some nucleosides]. AB - We studied the ability of phorbol 12-myristate 13-acetat to prevent erythroid differentiation and apoptosis in erythroleukemic K562 cells induced by cytidine, thymidine, and guanosine. The exposure of cancer cells to combinations of phorbol 12-myrsitate 13-acetate (100 nM) nucleosides for two days led to a loss of hemoglobin production (marker of erythroid differentiation) in cells and increased expression of monocyte-macrophage lineage associated surface antigen CD14. The treatment of K562 cells with nucleosides only was accompanied by the activation of caspase-3 and caspase-9, rather than caspase-6, increased fluorescence of ethidium bromide and DAPI upon binding to DNA, and apoptosis. Intracellular activation of caspase-6, inhibition of caspase-9, a markedly decreased activity of caspase-3 and of fluorescence of DNA-binding dyes, and inhibition of apoptosis were observed when the cells were treated with phorbol 12 myeristet 13-acetate combined with nucleosides. PMID- 15807434 TI - [Immunosuppressor activity of rat endometrial granulated cells and their differentiation]. AB - Natural killer and natural suppressor activities of the rat endometrial granulated cells were assayed on day 13 of pregnancy or pseudopregnancy. Metrial gland granulated cells were used as endometrial granulated cells. The natural killer activities of metrial gland granulated cells and other cells were determined by means of Hashimoto-Sudo test with K562 cells as targets. The estimation of natural killer activity included removal of the cells sticking to glass from a suspension of material gland granulated cells. Cytochemically, metrial gland granulated cells were identified by the presence of PAS-positive granules in the cytoplasm after treatment of the cells with diastase and identification of a specific antigen with the help of specific antisera. The natural killer activity of metrial gland granulated cells was twice weaker than that of splenocytes from the same pregnant or pseudopregnant females. The level of natural killer activity was proportional to the content of metrial gland granulated cells in a cell system. These data suggest that the natural killer activity of metrial gland granulated cells is realized via their contact with cell targets. Natural killer and suppressor activities were determined simultaneously for metrial gland granulated cells and splenocytes of the same rat with common cell targets. When estimating the nuclear suppressor activity of metrial gland granulated cells, the splenocytes of the same rat were used as an effector in a natural killer test. Various amounts of metrial gland granulated cells were added to the effector : target system at a ratio of 50:1. The natural suppressor activity of metrial gland granulated cells did not depend on the amount of metrial gland granulated cells present in a natural killer system. After fractionation in a Percoll gradient, the highest natural killer activity was recorded in a 30% Percoll fraction. The highest and lowest natural suppressor activities were recorded in 30% and 60% Percoll fractions, respectively. The culture medium was characterized by natural suppressor activity as well. The differences in mean areas of metrial gland granulated cells in 30 and 60% Percoll fractions between the pregnant (144.7 +/- 13.4 and 75.0 +/- 12.5 microm2, respectively) and pseudopregnant (97.5 +/- 4.9 and 69.2 +/- 3.5 microm2) females were reliable. The natural killer activity was estimated in all studied 23 samples of metrial gland granulated cells, among which 18 (79.6 +/- 7.8%) displayed the natural suppressor activity as well. The absence of natural suppressor activity in five samples was combined with the absence of this activity in their culture medium and with a reduction in the mean area of metrial gland granulated cells in 30% Percoll fraction to 109.1 +/- 5.2 microm2. The data obtained confirm the known data on a low activity of metrial gland granulated cells and demonstrated for the first time the natural suppressor activity of these cells. It was concluded that the natural suppressor activity of metrial gland granulated cells is due to their differentiation from metrial gland granulated cells with natural killer activity. PMID- 15807435 TI - [Regeneration of freshwater planarians Dugesia tigrina and Polycelis tenuis under the influence of methyl mercury compounds of natural origin]. AB - We studied the effects of methyl mercury compounds of natural origin on regeneration of the planarians Dugesia tigrina and Polycelis tenuis. Accumulation of methyl mercury in the planarian body leads to a delayed formation of photoreceptor organs in planarians of both species. After a significant traumatic load, the regeneration is suppressed and the death of some control and most experimental animals was observed. The intensity of joining additional cuts depends on the localization of body fragment with a cut and localization of a cut itself. PMID- 15807436 TI - [Structural organization and distribution of the symbiotic bacteria Wolbachia during spermatogenesis of Drosophila simulans]. AB - Electron microscopy and morphometric analysis have shown that the symbiotic bacteria Wolbachia occur the testis cells during spermatogenesis and are absent in mature spermatids. Bacteria did not affect the structural organization of testis cells, which have a typical morphology during morphogenesis. Bacteria were distributed along the meiotic spindle microtubules near the mitochondria. They increased in number in spermatids at the stage of elongation. Endosymbionts aggregated at the spermatid distal end and contained many vacuoles but were absent at the spermatid proximal end near the nuclei. It was shown for the first time that the diameter of spermatids in a strongly infected line was two of three times that in a noninfected line. We hypothesize that the increase in the number of endosymbionts during spermatid elongation can affect the chromatin condensation in the spermatozoon. PMID- 15807437 TI - [Changes in the distribution of mitochondria in mouse embryos blocked at the two cell stage]. AB - Changes in the distribution of mitochondria in the two-cell mouse embryos preceding the developmental arrest in vitro, caused by a genetically determined "two-cell block in vitro" or genisteine treatment, were examined vitally using the mitochondrial-specific probe rhodamine 123 and conventional fluorescence microscopy. In the former case, serious disturbances in the localization of mitochondria appeared already from the middle of two-cell stage, long before the time corresponding to the 2nd cleavage division. Comparison of the behavior of mitochondria in the embryos successfully developing between the one- and two-cell stages and that in the embryos that ceased to cleave suggests that the developmental arrest was accompanied by aggregation of the mitochondria into clusters. There are many such clusters unlike in the cytoplasm of normally developing embryos. Intracellular localization of clusters observed in the genisteine-treated embryos differed radically from that observed in the embryos blocked in vitro at the two-cell stage. PMID- 15807438 TI - [Comparative studies of follicle cells in testes of Glyptocephalus stelleri and Pleuronectes pinnifasciatus (Teleostei, Pleuronectidae)]. AB - Accessory cells were studied in early spermatogenesis of flatfishes Glyptocephalus stelleri and Pleuronectes pinnifasciatus using transmission electron microscopy. The morphological organization of accessory cells in G. stelleri was similar to that of Sertoli cells. In P. pinnifasciatus, these cells had morphological organization, which had not been previously described. PMID- 15807439 TI - [Neonatal androgenization of female rats during sex differentiation of the brain modifies neuronal organization of the amygdala complex]. AB - Sex differences in neuron dendroarchitectonics of the amygdala posterior cortical nucleus of adult rats were described for the first time using the Golgi method. Long-axon sparse-branched neurons in male rats possessed a larger number of primary dendrites, while female rats had long-axon dense-branched neurons with longer dendrites. Injection of testosterone propionate at 1250 g to females on day 5 after birth resulted in a greater number of primary dendrites of long-axon sparse branched neurons in adults, as compared to that in the control. Dendrites of long-axon sparse-branched neurons became much longer, thus enlarging the dendrite area. PMID- 15807441 TI - [Cholelithiasis: approaches to solving the problem]. PMID- 15807442 TI - [Low density lipoproteins in patients with cholelithiasis and cholesterosis of the gallbladder]. AB - AIM: To study heterogenic forms of LDLP and LP(a) in blood serum of patients with cholelithiasis (CL) and gallbladder cholesterosis (GBC). MATERIAL AND METHODS: Native gradient (3-12%) electrophoresis in polyacrylamide gel, rocket immunoelectrophoresis with antibodies to apo(a) were made in 20 patients with CL and 20 with GBC, 13 controls without gastrointestinal disease. Correlation of retardation factor (Rf) of LDLP and LP(a) with blood lipids, cholesterol (C) and triglycerides (TG) levels, body mass index (BMI) and age was studied. CL and GBC risk factors were analysed basing on a retrospective assessment in random representative samples of patients (100 CL and 100 GBC patients). RESULTS: There was a shift of the main peak in LDLP spectrum in the direction of smaller particles in GBC (Rf = 0.171 +/- 0.003) which was significant in comparison with CL group (Rf = 0.146 +/- 0.004, p < 0.001) and control (Rf = 0.114 +/- 0.013, p < 0.05). The analysis of LDLP Rf distribution in patients with different C levels has shown that LDLP small particles can occur in a normal C level: 75% in GBC and 50% in CL groups. Prevalence of small dense LDLP was recorded in both groups (87.5% cases) in hypercholesterolemia. Compared to control, LP(a) concentration was significantly elevated both in GBC (23.7 +/- 4.9 mg/dl) and CL (15.7 +/- 4.4 mg/dl) patients (control--7.5 +/- 1.4 mg/dl, p < 0.01), p > 0.5 in comparison between the groups. The correlation analysis found no correlations between LP9(a), other lipids, BMI and age in both study groups while Rf of LDLP correlated with C and TG levels (r = 0.596 and r = 0.226, respectively, p < 0.05), age and BMI (r = 0. 533 and r = 0.363, respectively, p < 0.05) in CL and did not correlate in GBC. CONCLUSION: A C level in CL changes with age and BMI while in GBC high LDLP C level was caused by other factors. No correlation of LP(a), LDLP Rf with age, body mass and blood lipids indicates that the above factors are independent in development of GBC. PMID- 15807443 TI - [Lipid composition of blood serum and bile in biliary diseases in patients with diabetes mellitus]. AB - AIM: To study content and correlation of lipids in the serum and bile from diabetic patients with biliary diseases (BD). MATERIAL AND METHODS: Serum and bile lipids were studied biochemically in 125 patients with BD and diabetes mellitus and 87 healthy control subjects aged 20-59 years. Serum lipids were studied biochemically as well as content and correlation of duodenal bile lipids. Correlation was investigated between blood and bile lipids. RESULTS: Compared to healthy controls, diabetic patients with chronic cholecystitis and cholelithiasis had higher serum total cholesterol, triglycerides (by 28-34% and 63%, respectively) and a 18-22% decrease in HDLP cholesterol. Bile of such patients is oversaturated with cholesterol. Patients with diabetes type 2 had a close direct correlation between levels of total cholesterol, LDLP in the serum and Thomas Hofmann index. CONCLUSION: BD patients with diabetes mellitus had high levels of serum lipids and high bile concentrations of cholesterol. Serum and bile lipids concentrations direcly correlate. PMID- 15807444 TI - [Use of essentiale and cholagogum in prevention of cholelithiasis in fat hepatosis with concomitant cholecystitis]. AB - AIM: To assess efficacy of essentiale in combination with cholagogum in prevention of cholelithogenesis in patients with fat hepatosis (FH) and concomitant cholecystitis (CC). MATERIAL AND METHODS: 100 patients aged 31-60 years with FH and CC were examined using clinical, device and biochemical tests. The diagnosis was verified by the results of ultrasonography, computed and MR tomography. Fractionated duodenal tubing followed by biochemical examination of bile was conducted in all the patients. RESULTS: Changes in physico-chemical properties of hepatic and vesical bile corresponding to stage I of cholelithiasis occurred in most of the examinees. Treatment with essentiale +cholagogum is clinically effective, improves functional condition of bile ducts and the liver, diminishes bile lithogenicity. CONCLUSION: In fat hepatosis and concomitant cholecystitis, essentiale and cholagogum improve biochemical composition of bile, resulting finally in decreased risk of cholelithogenesis. PMID- 15807445 TI - [Esomeprasol and rabeprasol effects on esophageal acidification in patients with gastroesophageal reflux disease, intensively metabolizing inhibitors of proton pump]. AB - AIM: To compare action of different esomeprasol and rabeprasol doses on esophageal acidification in patients with gastroesophageal reflux disease (GERD). MATERIAL AND METHODS: A cross-over study was made in 27 GERD patients (12 males and 15 females). It included esophagoduodenoscopy with biopsy for detection of H pylori and 24-h pH-metry of esophageal content. Rabeprasol and esomeprasol efficacy was compared for daily doses 20 mg of each (stage I) and rabeprasol 20 mg and esomeprasol 40 mg (stage II). RESULTS: All the patients proved H-pylori positive. Before the treatment mean daily time of marked esophageal acidification was 29-36%. At stage I significant differences between the drugs were absent. At stage If esomeprasol was more effective from the first day of therapy both in relation to time to effect and its intensity. CONCLUSION: Esomeprasol provided a fast and noticeable correction of esophageal acidification. This relieves symptoms of GERD and accelerates esophageal mucosa epithelization. PMID- 15807446 TI - [Relationship between CagA-bearing strains of Helicobacter pylori and gastrointestinal pathology]. AB - AIM: To study the prevalence of CagA-positive strains of Helicobacter pylori in Novosibirsk residents and to assess their relation to some clinical parameters. MATERIAL AND METHODS: Random sample of the population (218 males and 220 females aged 25-64 years) was examined. Antibodies against Helicobacter pylori and CagA protein were tested in the sera using enzyme immunoassay. Gastrointestinal symptoms were evaluated with questionnaire. RESULTS: The prevalence of H. pylori infection was 88%, among H. pylori-positive persons CagA-positivity was revealed in 58%. Presence of antibodies to CagA protein was associated with dyspepsia (OR = 2.5, 95% CI 1.2-2.5) and history of peptic ulcer (OR = 3.6, 95% CI 1.0-14.5). CagA-positive subjects more frequently reported bowel disturbances, diarrhea, in particular (OR = 2.9, 95% CI 1.2-7.4). CONCLUSION: The data showed similar distribution of H. pylori and CagA-positivity among adult population in Russia and Eastern European countries. CagA-positivity is associated with peptic ulcer and dyspepsia. PMID- 15807447 TI - [Impact of age and type of behavior on the course of ulcer disease]. AB - AIM: To study features of ulcer disease (UD) course with reference to a biological age, type of behavior, quality of life for recommendations on psychological rehabilitation of these patients. MATERIAL AND METHODS: Examination of 111 gastroduodenal UD patients determined type of behavior, correlation of biological age (BA) and due biological age (DBA), quality of life (QL). RESULTS: An unfavourable BA/DBA correlation was found in 71.1% cases, in which patients over 45 years of age had a larger ulcer defect size. Behavior type A with shorter scarring was observed in 30.9% patients. Lower QL was explained by sleep disorders, pain, emotional reactions, the demand in continuous treatment, diet, limited physical exercise. UD patients have a low QL index and a high index of the disease severity. These parameters are higher in unfavorable correlations between BA and DBA and in patients with type A behavior. CONCLUSION: UD patients have behavioral peculiarities dependent on BA and affecting the disease course and QL. PMID- 15807449 TI - [Quality of life and vegetative status of patients with ulcer disease]. AB - AIM: To analyse features of vegetative status of patients with ulcer disease (UD) with reference to quality of life (QL), course of UD, clinical and secretory motor characteristics. MATERIAL AND METHODS: A total of 200 UD patients were examined. UD was at the stage of exacerbation, new-onset, with a favourable or unfavourable course. The vegetative status was studied by parameters of Kerdo index, variation pulsometry, vegetative crises, insomnia. Quality of life (QL) was assessed by a medicosocial questionnaire. The patients were divided into two groups: with satisfactory and unsatisfactory QL. 120 patients had 24-h pH-metry. RESULTS: Among patients with normal QL normotonics and vagotonics prevailed. Low QL was seen in sympathicotonics. Sleep disorders and vegetative crises were in patients with vegetative imbalance, especially in combination with unsatisfactory QL. The worst QL was associated with sympathicotony. Vegetative balance was observed in a favourable UD course, moderate in time duodenogastric reflux (DGR). Sympathicotony runs with marked DGR, vasotony--minimal DGR, scare symptoms. CONCLUSION: Vegetative dysfunction in UD is due to permanent or paroxysmal vegetative disorders. Sympathicotony is accompanied with dyskinetic pain syndrome and longer duodenogastric reflux which is tolerated bad. Vegetative dystonia is more often in unfavorable course of UD. PMID- 15807448 TI - [Oxidative stress in mechanism of psychosomatic disorders realization in duodenal ulcer in students]. AB - AIM: To study the role of oxidative stress in mechanism of psychosomatic disorders involvement in pathogenesis of ulcerogenesis. MATERIAL AND METHODS: Psychosomatic disorders (PSD) and oxidative stress (OS) were assessed in 120 students with duodenal ulcer (DU) who had no concomitant pathology. Examination for PSD included interviewing by questionnaires. OS was judged by the level of lipid peroxidation products (malonic dialdehyde and dienic conjugates) and antioxidant activity--by glutathion peroxidase and superoxidedismutase. Melatonin was measured by enzyme immunoassay. RESULTS: A close correlation was discovered between the severity and features of PSD, lowering of melatonin, severity of OS and clinical features of the disease. CONCLUSION: DU students develop PSD which become leading factors of ulcerogenesis. One of the mechanisms of their participation in ulcerogenesis may be related with lowering of melatonin which is a potent stress regulator and corrector of lipid peroxidation and antioxidant activity. PMID- 15807450 TI - [Clinical variants of hypochondriac state in patients with duodenal ulcer]. AB - AIM: To describe some clinical variants of hypochondriac syndrome in patients with duodenal ulcer (DU). MATERIAL AND METHODS: A hypochondriac trend was studied clinically and was confirmed by changes in the first scale of AMPI in 150 DU patients. If the first scale contained a rise over 70 points, the examinee was stated to be much concerned about his/her health. RESULTS: Hypochondriac syndrome in exacerbation was registered in 59 (39.3%) patients. It was accompanied with anxiety in 28.8% cases. Introversion and original attitude to somatic disorders were observed in 25.4% cases. Demonstrative behavior was seen in 23.8% patients. Depressive trends occurred in 13.5% patients. Conception of the disease was confirmed in 5.1% cases. Hypochondriac state with protest reactions and hyperthymic effect occurred least frequently (1 and 1 case, respectively). CONCLUSION: Hypochondriac states aggravate DU, deteriorate quality of life. Early diagnosis of a variant of hypochondriac syndrome enables a directed individual psychotherapy which improves emotional state of the patient and prevent further development of psychopathological traits. PMID- 15807451 TI - [Choline- and histaminergic shifts as a trigger of gastroduodenal erosive-ulcer lesions in myocardial infarction]. AB - AIM: To clarify the role of cholin- and histaminergic shifts in the onset of circulatory and functional gastric disturbances associated with erosive-ulcer gastroduodenal lesions in myocardial infarction (MI). MATERIAL AND METHODS: Cholin- and histamine-reactive systems, end gastroduodenal mucosal circulation were studied in 80 Ml patients with gastroduodenal erosions and ulcers divided into three groups by the disease severity. RESULTS: MI features influence characteristics and severity of gastroduodenal erosions and ulcers. The latter develop due to focal microcirculatory disorders in gastric and duodenal mucosa by thrombohemorrhagic or thromboischemic type. Moderate and severe coronary pathology is associated with higher production of hydrochloric acid and pepsinogen, with depressed production of gastromucoproteins, hypo- or hypermotor dyskinesia, extremely severe coronary pathology--with decline of all gastric functions except normal acid production. Erosive-ulcer lesions may be triggered by alterations in cholin-histamine systems the activity of which in moderately severe and severe MI rises but in extremely severe--lowers. CONCLUSION: Pathogenesis of erosive-ulcer lesions of the gastroduodenal zone in MI has some specific features dependent on severity of MI. This should be taken in consideration in choice of drug therapy. PMID- 15807452 TI - [Efficacy of colloid bismuth subcitrate (De-Nol) in gastropathies induced by nonsteroid anti-inflammatory drugs: an open controlled 4-week trial]. AB - AIM: To evaluate efficiency of colloid bismuth subcitrate (CBS) in gastropathies induced by nonsteroid anti-inflammatory drugs (NSAID). MATERIAL AND METHODS: 45 patients with rheumatic diseases (RD) taking NSAID with gastric and/or duodenal up to 1 cm in size or multiple (more than 10) gastric mucosa erosions detected at esophagogastroduodenoscopy were randomized into two groups. Patients of group 1 (n = 30) received CBS (240 mg twice a day + amoxicillin 2 g/day and furozolidon 400 mg/day--subgroup 1a, n = 10 or CBS monotherapy in the same dose--subgroup 1b, n = 20). Patients of group 2 (n = 15) received ranitidine 150 mg twice a day. Older women, RA patients, patients with gastric ulcer prevailed. H. pylori was detected in 73.3 and 90% patients, respectively. Dispepsia and heartburn occurred in 90 and 93.3%, respectively. The efficacy was assessed after 4 weeks of therapy. RESULTS: Healing of ulcers and erosions were achieved in 22 of 26 patients of group 1 (84.6%) and in 7 of 14 (50%) of group 2 (p = 0.036, chi square). After therapy dyspepsia persisted in 4 patients of group 1 and 9 patients of group 2. Side effects were in 3 patients on CBS. Untiulcer effect did not differ between the subgroups (88.8 and 82.4%, respectively) and was effective in 6 of 7 HP-negative patients (85.7%) and in 8 of 10 HP-positive patients (80%). CONCLUSION: De-Nol is effective in NSAID-induced gastropathies. Its action is not related with influence on HP. PMID- 15807453 TI - [Current understanding of primary biliary cirrhosis]. AB - AIM: To examine characteristics of the course of primary biliary cirrhosis (PBC). MATERIAL AND METHODS: The study of 150 PBC patients aged 26-82 years used clinical, biochemical, immunological and morphological methods. RESULTS: Antimitochondrial antibodies of type M2 (AMAM2) were detected in 85.4% patients with PBC. A positive correlation was found between clinicobiochemical activity of the disease and number of AMAM2. AMAM2-positive patients more frequently had extrahepatic manifestations, overlap syndrome, higher biochemical and immunological activity vs AMAM2-negative patients. CONCLUSION: The following variants of PBC course were identified: classic AMAM2-positive, AMAM2-negative, overlapping of PBC and autoimmune hepatitis. Concentration of AMAM2 has diagnostic and prognostic value in PBC. PMID- 15807454 TI - [Immunogenetic HLA markers of chronic viral hepatitis]. AB - AIM: To study possible immunogenetic HLA markers of chronic viral hepatitides. MATERIAL AND METHODS: Using the reaction of complement-dependent cytotoxicity by Terasaki, we analysed distribution of leukocytic HLA antigens (loci A, B and C) in 179 patients with chronic viral hepatitides B, C and D in Russians and Kazakhs living in the Astrakhan Region. RESULTS: In the Russian population we discovered a significant positive association of CVHB with HLA-B18, HLA-B35, HLA-B40, HLA Cw3 antigens, and negative one--with HLA-A2. In Kazakhs with CVHB there was a positive association with HLA-A3, HLA-B18 and negative one--with HLA-A11. Alleles HLA-A10, HLA-B35, HLA-B40 and HLA-Cw3 mark CVHC in Russians. HLA-Cw4 specificity acts as protector in development of chronic HCV-infection. A correlation was found between carriage of some specificities and haplotypes of HLA and activity of chronic HBV and HCV infection. A high risk of chronic delta infection in Russians is associated with HLA-B8 and HLA-B35, in Kazakhs--with HLA-B35 and HLA D40. There are significant associations between CVHB, CVHC, chronic delta infection and some HLA haplotypes. CONCLUSION: A universal role of HLA-B35 specificity in development of CVH irrespective of hepatotropic virus and patients' nationality is shown. PMID- 15807455 TI - [Spectrum, incidence and prognostic value of systemic manifestations in chronic hepatitis C of low activity]. AB - AIM: To study the incidence and prognostic significance of extrahepatic manifestations of chronic HCV-infection with low activity of hepatic process and to specify treatment policy. MATERIAL AND METHODS: Sixty-two patients with low active chronic hepatitis C (CHC) entered the study. Blood samples from all the patients were studied clinically, biochemically, immunologically and virusologically. In 58 patients the diagnosis was confirmed morphologically (liver tissue). RESULTS: Systemic manifestations of CHC were observed in 23 (37.1%) patients, in 12 of them they were associated with cryoglobulinemia. Most frequent was cryoglobulinemic vasculitis with skin (12.9%) and joint (9.7%) involvement, cryoglobulinemic glomerulonephritis (11.3%), arthralgia without association with cryoglobulinemia (11.3%). Independent risk factors for development of systemic disorders in CHC of low activity comprised female gender, age over 45, the disease duration and presence of cryoglobulinemia. CONCLUSION: CHC of low activity is characterized by high occurrence of systemic manifestations primarily associated with cryoglobulinemia. Renal affection with development of cryoglobulinemic glomerulonephritis is most prognostically significant. PMID- 15807456 TI - [The condition of the stomach, duodenum and bile ducts in patients with chronic hepatitis C]. AB - AIM: To study the status of the stomach, duodenum and biliary tracts in young patients with chronic hepatitis C. MATERIAL AND METHODS: We studied associated pathology of the gastroduodenal zone and biliary tract in 140 patients with CHC aged 16 to 35 years (mean age 25 +/- 0.8 years). We used USI and MRT of the abdominal organs, fibroesophagogastroduodenoscopy with biopsy of gastric mucosa for H. pylori, fractional duodenal tubing. 30 healthy males were controls. RESULTS: Asymtomatic gastroduodenal and biliary defects were found (75.5%). Endoscopic signs of gastroduodenal lesions were frequent (76.3%) in CHC patients, the majority of them were associated with H. Pylori and were not significantly different from those in the controls. Biliary tracts pathology, as shown by USI and MRT, occurred in 50.8%, were primarily functional (74.5%) and combined with high occurrence of bile mixed microflora with predominance of B. Fragilis (81%). Correlation of pathological changes of the gastroduodenal zone and biliary tract was established in CHC patients. How ever, they do not correlate with clinicolaboratory and histological signs of HCV infection. CONCLUSION. Young patients with CHC demonstrated no correlation between pathological changes of the gastroduodenal zone, biliary tracts and clinicolaboratory, histological signs of HCV infection. Therefore, it is recommended not to eradicate HP-infection and not to suppress opportunistic bile microflora in CHC young patients in the absence of clinical symptoms of gastrointestinal changes. PMID- 15807457 TI - [Cytokine spectrum in patients with chronic hepatitis C in the treatment with interferon-alpha drugs]. AB - AIM: To evaluate the trend in content of IL-1, 2, 4, 6, TNF-alpha, interferon alpha (Inf-alpha) and gamma (Inf-g) in the serum of patients with chronic hepatitis C (CHC) in the course of treatment with Inf-alpha. MATERIAL AND METHODS: The study included 30 patients with CHC (20 males and 10 females) and 25 healthy controls. Markers of CHC were detected with enzyme immunoassay, RNA--with polymerase chain reaction. Index of histological activity was calculated for 20 patients. Immune response mediators were determined with enzyme immunoassay before therapy with intron A in a dose 3000000 U 3 times a week and after 12 weeks of such treatment. RESULTS: Before the treatment absolute levels of inflammation mediators were high, after 3 months of treatment RNA of VHC was absent in 16 (55%) of the patients, 18 (60%) had low activity of transaminases. In 6 months VHC-RNA was not detected in 8 (28%) patients. Patients with a persistent positive effect had Ifn-g significantly higher than in unstable effect. IL-4 showed an opposite trend. CONCLUSION: High concentrations of IL-4 and low of Ifn-g in the serum of CHC patients before If treatment determined high activity of Th-2 system mediators, active replication of HCV and worse response to therapy. PMID- 15807458 TI - [Treatment with peg-interferon-alpha of patients with chronic hepatitis C]. AB - AIM: To investigate a comparative efficacy of treatment with different types of interferon. MATERIAL AND METHODS: We studied comparative effectiveness of alpha interferon vs pegilated interferon. In both cases alpha-interferon was administered in combination with ribavirin. 98 patients (age 16-65 years) with chronic HCV infection have undergone serological, virological, biochemical and histological tests for diagnosis and characterization of HCV infection. HCV genotyping was done by sequencing-based technology. RESULTS: It is shown that treatment with pegilated interferon-alpha in combination with ribavirin resulted in a higher rate of sustained viral response. Adverse reactions were less frequent with peg-interferon vs conventional interferon therapy. CONCLUSION: Treatment with peg-interferon-alpha in combination with ribavirin is more effective than conventional interferon therapy. PMID- 15807459 TI - [Combined treatment with reaferon and iodantipirin in patients with chronic viral hepatitides B and C]. AB - AIM: To examine clinical efficacy of combined therapy with reaferon and iodantipirin in patients with chronic hepatitis B and C of moderate activity. MATERIAL AND METHODS: 100 patients with confirmed diagnosis of chronic viral hepatitis (CVH) at the stage of replication. All the patients have undergone puncture biopsy of the liver with estimation of index of histological activity Knodell and index of fibrosis Desmeth. RESULTS: Comparative evaluation was made of clinical efficacy of combined therapy reaferon + iodantipirin and monotherapy with reaferon in 100 patients with moderate activity CHB and C. CONCLUSION: Conventional scheme with reaferon turned out less effective than the above combination therapy. PMID- 15807460 TI - [Alcoholic cholangitis as a variant of alcoholic disease of the liver (case report and review of the literature)]. PMID- 15807461 TI - [Microscopic colitis--a cause of a torpid course of celiac disease]. PMID- 15807462 TI - [Collagenic sprue as a cause of dynamic obstruction of the small intestine]. PMID- 15807463 TI - [Dynamic results of the EUROPA study: mechanisms of a favourable influence of perindopril on the course of IHD and decreased IHD mortality]. PMID- 15807464 TI - [Diagnosis and conservative treatment of acute pyelonephritis in diabetes mellitus]. AB - AIM: To raise efficacy of conservative treatment of patients with acute pyelonephritis (AP) which developed in the presence of diabetes mellitus (DM). MATERIAL AND METHODS: Elderly patients (n = 218, 182 females and 36 males) with DM and AP enered the study. DM type 1 and 2 were diagnosed in 41 and 177 of them. Pyelonephritis was diffuse-purulent, pyodestructive, calculous, pyocalculous. RESULTS: Conservative treatment was made in 160 patients, surgical treatment--in 58. Lethal outcomes (14 cases) were registered as a rule in patients with bilateral pyodestructive pyelonephritis. CONCLUSION: AP is a severe complication of DM. The severity of DM decompensation and metabolic disorders is proportional to AP severity. AP in diebetics runs often an asymptomatic course and the diagnosis is difficult. Therapeutic policy is individual with consideration of a clinical course. Positive results were achieved in 80% patients. PMID- 15807465 TI - [Female sexual hormones and some other factors in pathogenesis of cholelithiasis]. PMID- 15807467 TI - Congressional budgeters want tougher Medicaid cuts than Bush. PMID- 15807466 TI - [Ghrelin and peptide YY--regulators of the appetite and quantity of taken food. Perspectives of cachexia and obesity treatment]. PMID- 15807468 TI - Howard Atwood Kelly, Elizabeth Hurdon, and their great work: "The vermiform appendix and its diseases". PMID- 15807469 TI - Appendicitis and infections of the appendix. AB - The pathologic spectrum of the acutely inflamed appendix encompasses a wide range of infectious and noninfectious entities. The appendix suffers alone in some of these disorders, and in others may be involved through extension from other areas of the gastrointestinal tract. Although the appendix is the most commonly resected and examined intraabdominal organ, the pathogenesis and etiology of acute nonspecific appendicitis (the most common diagnosis made in this organ) remains enigmatic. This review encompasses the pathology, pathogenesis, and bacteriology of acute appendicitis, as well as controversial issues such as the diagnosis of chronic appendicitis and the significance of a morphologically unremarkable appendectomy specimen in the clinical context of appendicitis. In addition, the pathologic features, pertinent diagnostic techniques, and clinical significance of several specific bacterial, viral, fungal, and parasitic infections affecting the appendix are presented, including adenovirus, cytomegalovirus, Yersinia species, actinomycosis, Mycobacteria species, histoplasmosis, pinworms, schistosomiasis, and Strongyloides stercoralis. PMID- 15807470 TI - Granulomatous appendicitis and the appendix in idiopathic inflammatory bowel disease. AB - Granulomatous appendicitis is a rare condition, accounting for less than 2% of all cases of appendicitis. The initial belief that it represented a manifestation of Crohn's disease is incorrect in the great majority of cases, as only 5-10% of patients with granulomatous appendicitis develop Crohn's disease elsewhere in their gastrointestinal tract. The remaining etiologies are diverse. Unusual causes include sarcoidosis, foreign body reaction, and infection by mycobacteria, fungi, or parasites. These conditions combined explain less than 10% of cases. More recently, two etiologies have been recognized that potentially account for most of the previous "idiopathic" cases of granulomatous appendicitis. The first is infection by pathogenic Yersinia species, now demonstrated in approximately 25% of cases. The second cause may be the most common of all, namely subacute/recurrent appendicitis with interval appendectomy. This condition likely produces a granulomatous reaction in relation to a protracted secondary inflammatory response to appendicitis and temporizing measures to delay appendectomy, such as antibiotic therapy. Thus, granulomatous appendicitis only rarely represents a manifestation of Crohn's disease. Rather, the overwhelming majority of patients with this condition are cured by appendectomy alone. The appendix, however, can be involved by idiopathic inflammatory bowel disease, both Crohn's disease and ulcerative colitis. It can be involved by ulcerative colitis in patients with distal colonic involvement and sparing of the intervening colonic segment, a phenomenon known as the appendiceal "skip lesion" or "cecal patch" and this pattern of involvement does not necessarily indicate Crohn's disease. Interestingly, appendectomy has been shown to provide some protection against developing inflammatory bowel disease and in reducing its severity if performed before the onset of disease. PMID- 15807471 TI - Neuroendocrine tumors of the appendix. AB - The vast majority of neuroendocrine neoplasms of the appendix are carcinoid tumors. Most are of enterochromaffin (EC) cell type, although rare examples are of L cell type. EC cell carcinoids of the appendix differ from those encountered elsewhere in the gastrointestinal system. For example, they are remarkably common given the small size of the appendix, are usually benign, occur in younger patients, and typically contain sustentacular cells that express S-100. Origin from subepithelial neuroendocrine cells could explain these characteristics. It has also been suggested that most appendiceal carcinoids are hyperplastic rather than neoplastic, although this hypothesis requires further study. Nevertheless, truly neoplastic EC cell carcinoids of the appendix undoubtedly occur, and those greater than 2 cm in diameter have a significant risk of producing distant metastases. Carcinoid syndrome is a very rare presentation. Tubular carcinoids are unusual benign neoplasms; it has been proposed that they represent L cell carcinoids with a predominant tubular pattern of growth. Goblet cell carcinoids tend not to produce a grossly visible tumor mass but diffusely infiltrate the wall. They typically exhibit tight clusters of goblet cells, usually with scattered neuroendocrine cells and sometimes with Paneth cells, sometimes surrounding a small lumen. They may behave as a low-grade malignancy. The distinction between goblet cell carcinoid and other types of tumor is of great importance because of the implications for treatment and prognosis. Frank adenocarcinoma can arise from goblet cell carcinoids, and tumors with both components are classified as mixed goblet cell carcinoid-adenocarcinoma. The carcinoma component of the latter determines their prognosis, which would be worse than for a goblet cell carcinoid alone. PMID- 15807472 TI - Primary epithelial neoplasms and other epithelial lesions of the appendix (excluding carcinoid tumors). AB - Epithelial tumors of the appendix range from low-grade mucosal-based tumors which, when confined to the appendix, have an excellent prognosis but, once outside the appendix, have a fair prognosis and often a prolonged disease course, to high grade invasive carcinomas that are rapidly fatal. Low grade mucinous neoplasms may rupture and spread to the peritoneum as pseudomyxoma peritonei, and the nomenclature of these tumors has been the subject of considerable disagreement among pathologists; the designation "low grade appendiceal mucinous neoplasm" has recently been proposed for reasons discussed herein. Demonstrating rupture of these neoplasms may require particularly diligent gross and microscopic evaluation as the rupture site often heals over leaving only subtle evidence of its presence. Invasive adenocarcinomas are often mucinous and may also spread to the peritoneum. Against this backdrop, the clinical and pathologic features of low grade appendiceal mucinous neoplasms and mucinous adenocarcinomas, as well as other types such as typical colorectal type and signet-ring cell type, are reviewed. In addition, emerging entities, serrated polyps and serrated adenomas, whose significance is only beginning to be understood, are considered. Retention cysts, hyperplastic polyps, and diffuse mucosal hyperplasia, although not necessarily neoplastic, are reviewed here as they may enter into the differential diagnosis of appendiceal mucinous neoplasms. PMID- 15807473 TI - Pseudomyxoma peritonei and selected other aspects of the spread of appendiceal neoplasms. AB - The varied aspects of spread of appendiceal neoplasms are reviewed with emphasis on the often clinically dramatic phenomenon known as pseudomyxoma peritonei, a term mostly used to describe grossly evident mucin within the peritoneal cavity. The majority of cases of pseudomyxoma peritonei result from tumors primary in the appendix, which are usually low-grade. On microscopic examination pseudomyxoma peritonei is typically characterized by large aggregates of mucin which may be relatively acellular or cellular containing strips of mucinous epithelium, mucinous epithelium encircling glands and cysts, or aggregates of mucinous epithelium lying within mucin pools. High-grade adenocarcinoma of the appendix may spread to the omentum and peritoneal surfaces without grossly striking mucin deposition and resemble spread of other high-grade gastrointestinal adenocarcinomas. In many cases of pseudomyxoma peritonei in females there is involvement of one, or more often, both ovaries. The size of the ovarian neoplasms characteristically dwarfs the often relatively unremarkable appearing appendix in these cases. The ovaries are typically multilocular, although one locule may dominate, and in cases in which the primary is a low-grade appendiceal mucinous neoplasm often have a "jelly-like" consistency. In cases of spread of frank adenocarcinomas the ovarian metastases typically have a more solid, albeit still somewhat gelatinous consistency. Microscopic examination of the ovaries typically shows surface involvement, a characteristic of spread to the ovaries in general, and the glands and cysts that replace most or all of the parenchyma are typically lined by tall mucin-rich cells with, in many cases, relatively bland microscopic features. In cases of frank adenocarcinoma, the tumors may mimic closely a primary mucinous adenocarcinoma of the ovary. Spread to the ovaries may also be seen in cases of frank intestinal-type adenocarcinoma primary in the appendix and the uncommon signet ring cell carcinoma of the appendix, the latter being one cause of the Krukenberg tumor. Occasional cases are reported in the literature of ovarian spread of goblet cell carcinoid tumor of the appendix, but in our opinion most of the primary tumors in those cases are better classified as adenocarcinomas, usually dominantly of signet-ring cell type, albeit sometimes with focal neuroendocrine differentiation. Other interesting aspects of spread of appendiceal neoplasms include to the lining of the uterus and the fallopian tube. In yet other cases the tumors may present clinically as incidentally discovered mucinous aggregates within hernia sac specimens or as a scrotal mass. PMID- 15807474 TI - Miscellaneous conditions of the appendix. AB - A variety of miscellaneous conditions affect the appendix, both as incidental findings and as causes of clinical signs and symptoms that often mimic appendicitis. Congenital abnormalities of the appendix are rare; the two most commonly reported are congenital absence and appendiceal duplication. Diverticular disease may be an incidental finding, but when inflamed, can be clinically confused with appendicitis. Endometriosis of the appendix, which usually occurs in the setting of generalized gastrointestinal endometriosis, often presents as acute appendicitis, but may present as intussusception, lower intestinal bleeding, and, particularly during pregnancy, perforation. Peritoneal endosalpingiosis often involves the appendiceal serosa and occasionally the wall but has no clinical manifestations in contrast to endometriosis. Vasculitis may be either isolated to the appendix or part of a systemic vasculitis, most often polyarteritis nodosa. Neural proliferations of the appendix include lesions associated with von Recklinghausen's disease, as well as mucosal and axial neuromas that are theorized to progress to fibrous obliteration of the appendix. Mesenchymal tumors of the appendix are most often of smooth muscle type, usually leiomyoma but rarely leiomyosarcoma; nonmyogenic neoplasms such as gastrointestinal stromal tumor, granular cell tumor, Kaposi's sarcoma, and miscellaneous other curiosities occur rarely. Lymphoma affects the appendix exceptionally; in children, Burkitt lymphoma is most common whereas in adults, large cell lymphomas and low grade B-cell lymphomas predominate. Secondary involvement of the appendix by leukemia has been reported. Secondary involvement of the appendix by carcinomas of the female genital tract, particularly ovary, and diverse other sites are in aggregate common but only rarely a clinical or pathological difficulty. Occasionally, however, appendiceal neoplasia that is secondary from another site may dominate the clinical picture and lead to potential pathologic misdiagnosis as primary appendiceal disease. PMID- 15807475 TI - The guidelines for prevention and control of overweight and obesity in Chinese adults. PMID- 15807476 TI - Refraction status and motility defects of persons 4-74 years. PMID- 15807481 TI - On the perception of molecules from 3D atomic coordinates. AB - A method is presented for perceiving chemical types of atoms in molecules given 3D atomic coordinates and element identities. The method assigns hybridizations, bond orders, and formal charges for structures whether hydrogen atoms are present. The Maximum Weighted Matching algorithm for nonbipartite graphs is used to assign bond orders with weights derived from statistics of a large collection of organic molecules. Results form tests on a collection of functional groups, heterocycles, entries from the Protein Data Bank, and Cambridge Structural Database as well as a comparison to other methods, are presented and discussed. PMID- 15807482 TI - Variable neighborhood search for extremal graphs. 10. Comparison of irregularity indices for chemical trees. AB - Chemical graphs, as other ones, are regular if all their vertices have the same degree. Otherwise, they are irregular, and it is of interest to measure their irregularity both for descriptive purposes and for QSAR/QSPR studies. Three indices have been proposed in the literature for that purpose: those of Collatz Sinogowitz, of Albertson, and of Bell's variance of degrees. We study their properties for the case of chemical trees. Structural conjectures are generated with the system AutoGraphiX, and most of them proved later by mathematical means. Analytical expressions for extremal values are obtained, and extremal graphs are characterized for the two last indices. PMID- 15807483 TI - From Wiener index to molecules. AB - In this paper we present an algorithm for the generation of molecular graphs with a given value of the Wiener index. The high number of graphs for a given value of the Wiener index is reduced thanks to the application of a set of heuristics taking into account the structural characteristics of the molecules. The selection of parameters as the interval of values for the Wiener index, the diversity and occurrence of atoms and bonds, the size and number of cycles, and the presence of structural patterns guide the processing of the heuristics generating molecular graphs with a considerable saving in computational cost. The modularity in the design of the algorithm allows it to be used as a pattern for the development of other algorithms based on different topological invariants, which allow for its use in areas of interest, say as involving combinatorial databases and screening in chemical databases. PMID- 15807484 TI - Prospective exploration of synthetically feasible, medicinally relevant chemical space. AB - We describe a novel approach to direct the exploration of chemical space in an effort to balance synthetic accessibility and medicinal relevancy prior to experimental work. Reaction transforms containing empirical reactivity and compatibility information are dynamically assembled into reaction sequences (vProtocols) utilizing commercially available starting material feedstock. These vProtocols are evolved and optimized by a genetic algorithm, which leverages fitness functions based on predicted properties of generated molecular products. We present the underlying concepts, methodology and initial results of this prospective approach. PMID- 15807485 TI - Classifying 'drug-likeness' with kernel-based learning methods. AB - In this article we report about a successful application of modern machine learning technology, namely Support Vector Machines, to the problem of assessing the 'drug-likeness' of a chemical from a given set of descriptors of the substance. We were able to drastically improve the recent result by Byvatov et al. (2003) on this task and achieved an error rate of about 7% on unseen compounds using Support Vector Machines. We see a very high potential of such machine learning techniques for a variety of computational chemistry problems that occur in the drug discovery and drug design process. PMID- 15807486 TI - A partition coefficient calculation method with the SFED model. AB - The Solvation Free Energy Density (SFED) model, a solvation model proposed by No et al. was modified to give better solvation free energies of the molecules having high polarizable groups. The SFED at a point around the molecule was represented by a linear combination of four basis functions, the contribution from the cavitation free energy of a solvent, and a constant. As an application of the SFED model, the linear expansion coefficients of the Hydration Free Energy Density (HFED) and the 1-Octanol Free Energy Density (1-OFED) were determined. Both calculated hydration free energy and 1-octanol solvation free energy of selected 95 organic molecules agreed well with experimental values. The standard errors were 0.47 and 0.39 kcal/mol, respectively. 1-Octanol/water partition coefficients (P) of the molecules were calculated from the difference of the HFE and 1-OFE of the molecules. At the same time, the logP density (LPD) of a molecule was represented by the same basis functional form with the SFED model. The logP of a molecule can be obtained by the integration of the LPD of the molecule. The coefficients of the basis functions were determined by using experimental logP as constraints through an optimization procedure. Both logPs calculated from the free energy difference and from the LPD agreed well with the experimental data. The absolute mean errors were obtained as 0.34 and 0.32, respectively. PMID- 15807487 TI - Kohonen network study of aromatic compounds based on electronic and nonelectronic structure descriptors. AB - Atoms in Molecules (AIM) and Electron Localization Function (ELF) methodologies were applied to describe the electronic structure of 88 aromatic compounds. The analyzed database contains molecules substituted by nucleophilic and electrophilic groups which are responsible for electron density distribution in the molecule and further for its reactivity. Radial Distribution Function (RDF), Weighted Holistic Invariant Molecular (WHIM), Three-Dimensional Molecule Representation of Structures based on Electron Diffraction (3D-MoRSE) and Geometry, Topology and Atom-Weights Assembly (GETAWAY) descriptors were taken into account describing the structures of the analyzed molecules. According to generated descriptor space the classification of the molecules has been subsequently performed using unsupervised learning strategy and Kohonen network. The final step of descriptor space testing was supervised learning of Counter Propagation Artificial Neural Network (CPANN) using n-octanol/water partition coefficient (logP), dipole moment (DM) and molecular refractivity (MR) as target values. PMID- 15807488 TI - Do Fukui function maxima relate to sites of metabolism? A critical case study. AB - The usefulness of local reactivity descriptors for understanding drug metabolism is investigated. Electrophilic Fukui functions are calculated for 18 drugs and 11 agrochemicals and their relation to experimentally observed metabolites is discussed. Maxima of the Fukui functions correspond to major sites of metabolic attack in many examples, facilitating a posteriori understanding of experimental findings. In the second part of the paper, the nature of the electrophilic oxidant species in cytochromes, called "Compound I" (Cpd I), is investigated within the Fukui framework. Nucleophilic Fukui functions are calculated involving the relevant spin states of Cpd I, allowing a more qualitative, intuitive understanding of its reactivity. PMID- 15807490 TI - Periodic cages. AB - Various cages are constructed by using three types of caps: f-cap (derived from spherical fullerenes by deleting zones of various size), kf-cap (obtainable by cutting off the polar ring, of size k), and t-cap ("tubercule"-cap). Building ways are presented, some of them being possible isomerization routes in the real chemistry of fullerenes. Periodic cages with ((5,7)3) covering are modeled, and their constitutive typing enumeration is given. Spectral data revealed some electronic periodicity in fullerene clusters. Semiempirical and strain energy calculations complete their characterization. PMID- 15807489 TI - 2D-3D migration of large chemical inventories with conformational multiplication. Application of the genetic algorithm. AB - Mathematical chemistry has afforded a variety of research areas with important tools to understand and predict the behavior of chemicals without having to consider the complexities of three-dimensional conformations of molecules. Predictive toxicology, an area of increasing importance to toxicity assessments critical to molecular design and risk management, must be based on more explicit descriptions of structure, however. Minimum energy conformations are often used for convenience due, in part, to the difficulty of computing a representative population of conformers in all but rigid structures. Such simplifying assumptions fail to reveal the variance of the stereoelectronic nature of molecules as well as the misclassification of chemicals which initiate receptor based toxicity pathways. Because these errors impact both the success in discovering new lead and the identification of possible hazards, it is important that mathematical chemistry develop additional tools for conformational analysis. This paper presents a new system for automated 2D-3D migration of chemicals in large databases with conformer multiplication. The main advantages of this system are its straightforward performance, reasonable execution time, simplicity and applicability to building large 3D chemical inventories. The module for conformer multiplication within the 2D-3D migration system is based on a new formulation of the genetic algorithm for computing populations of possible conformers. The performance of the automated 2D-3D migration system in building a centralized 3D database for all chemicals in commerce worldwide is discussed. The applicability of the 3D database in assessing the impact of molecular flexibility on identifying active conformers in QSAR analysis and assessing similarity between chemicals is illustrated. PMID- 15807491 TI - Structure, dynamics and solvation of HIV-1 protease/saquinavir complex in aqueous solution and their contributions to drug resistance: molecular dynamic simulations. AB - As it is known that the understanding of the basic properties of the enzyme/inhibitor complex leads directly to enhancing the capability in drug designing and drug discovery. Molecular dynamics simulations have been performed to examine detailed information on the structure and dynamical properties of the HIV-1 PR complexed with saquinavir in the three protonated states, monoprotonates at Asp25 (Mono-25) and Asp25'(Mono-25') and diprotonate (Di-Pro) at both Asp25 and Asp25'. The obtained results support clinical data which reveal that Ile84 and Gly48 are two of the most frequent residues where mutation toward a protease inhibitor takes place. In contrast to the Ile84 mutation due to high displacement of Ile84 in the presence of saquinavir, source of the Gly48 mutation was observed to be due to the limited space in the HIV-1 PR pocket. The Gly48 was, on one side, found to form strong hydrogen bonds with saquinavir, while on the other side this residue was repelled by the hydrophobic Phe53 residue. In terms of inhibitor/enzyme binding, interactions between saquinavir and a catalytic triad of the HIV-1 PR were calculated using the ab initio method. The results show an order of the binding energy of Mono25